PMID- 8220520 TI - Conceptual framework for nutrition surveillance systems. AB - This article describes the evolution of nutrition surveillance as an intervention strategy and presents a framework for improving the usefulness of nutrition surveillance programs. It seems clear that such programs' impact on nutritional well-being will depend increasingly on their ability to reach and influence decision-makers. Therefore, it is important to consider political and social forces, and also to realize that if a program is too decentralized or too far removed from key decision-makers, its ability to influence resource flows may be limited. It is of course important that the surveillance information provided be appropriate and of good quality. Therefore, the data collected should be analyzed to ensure they are accurate and representative. Once that has been done, relevant findings should be presented in a readily understandable form designed to meet the intended recipients' information needs. Such findings should also be disseminated to all important decision-maker constituencies, including external donors of nutrition assistance and the general public. PMID- 8220521 TI - Chagas' disease: control and surveillance through use of insecticides and community participation in Mambai, Goias, Brazil. AB - This article describes the nature and effectiveness of methods developed for longitudinal surveillance and control of Chagas' disease vectors in Mambai, Brazil. The surveillance effort made use of an education program in the schools, community leader and resident participation, specific surveillance equipment (Gomez-Nunez boxes, instructive posters, and instructive calendars with special detection papers), periodic manual collection of triatomines, and a network of collection posts. Spraying of infested houses was accomplished partly by community leaders who received special training for this purpose. The methods used offer an affordable approach to triatomine surveillance and control well suited to use in large-scale national programs. PMID- 8220522 TI - Global malaria control strategy. PMID- 8220523 TI - Investing in health: the 1993 World Development Report of the World Bank. PMID- 8220524 TI - Strategies for the certification of the eradication of wild poliovirus transmission in the Americas. Expanded Program on Immunization, Pan American Health Organization. AB - Because it appears that the last case of poliomyelitis caused by transmission of indigenous wild poliovirus occurred 2 years ago on 23 August 1991 in Peru, the challenge for PAHO is to develop a methodology of certification that will convince the rest of the world that the eradication of poliomyelitis in the Americas has been achieved. To that end, the guidelines set forth here allow for a rigorous and standardized evaluation of the certification process by objective experts. As it was during the campaign to eradicate smallpox, epidemiologic surveillance (of acute flaccid paralysis [AFP] in the case of polio) will be the most important component of the certification process. It must be demonstrated that when cases of AFP occur, they will be identified, reported, and investigated in a timely manner to insure that if wild poliovirus is present, it will be identified. PMID- 8220525 TI - Association between HIV and tuberculosis: technical guide. PMID- 8220526 TI - [How do chronic leg ulcers originate?]. PMID- 8220527 TI - [Chronic ulcers can be treated]. PMID- 8220528 TI - [Compression can prevent ulcers]. PMID- 8220529 TI - [Living with psoriasis--a qualitative study]. PMID- 8220530 TI - [A place to be or a place to learn?]. PMID- 8220531 TI - [Student guidance. Do as I say but not as I do!]. PMID- 8220532 TI - [Current therapy of birth marks]. PMID- 8220533 TI - [Sterile center--an important part of quality assurance]. PMID- 8220534 TI - [What viewpoint of knowledge does the journal Sykepleien provide?]. PMID- 8220535 TI - [AIDS in the world]. PMID- 8220537 TI - [Continued education provides new impetus]. PMID- 8220538 TI - [Foreign nurses]. PMID- 8220536 TI - [We foreign nurses]. PMID- 8220539 TI - [Why I gave birth at home!]. PMID- 8220540 TI - [Closing-time routines in neonatal intensive care units]. PMID- 8220541 TI - [Rosenborg Project: nursing home residents with senile dementia--hospital report]. PMID- 8220542 TI - [The spiritual dimension of nursing]. PMID- 8220543 TI - [Dry treatment best for diabetic ulcers]. PMID- 8220544 TI - [Bechterew's disease and Reiter's disease]. PMID- 8220545 TI - [Nurses in rheuma-care]. PMID- 8220546 TI - [Systemic lupus erythematosus]. PMID- 8220547 TI - [Polymyalgia rheumatica]. PMID- 8220548 TI - [Rheuma surgery and nursing care]. PMID- 8220549 TI - [Rheumatic diseases. Child and adolescent department in the hospital]. PMID- 8220551 TI - [Rheumatic diseases. Discussion groups]. PMID- 8220550 TI - [Rheumatic diseases. Current treatment methods]. PMID- 8220552 TI - [A 24-hour open nursing conference]. PMID- 8220553 TI - [Deaconess House Lovisenberg. 125 years for parish and nursing]. PMID- 8220554 TI - [Nursing services and DRG]. PMID- 8220555 TI - [Rheumatic diseases. A life with arthritis--2 patient histories. Interview by Tor Egil Thomassen and Ellen Hunstad]. PMID- 8220556 TI - [The oldest old and life's meaning]. PMID- 8220557 TI - [Rosenborg Project: nursing home residents with senile dementia--treatment methods in family therapy]. PMID- 8220558 TI - [Does Kari Martinsen's work have clinical relevance?]. PMID- 8220559 TI - [Nursing college and research]. PMID- 8220560 TI - [Who considers the patients?]. PMID- 8220562 TI - [Yes, to operating room practice in basic education]. PMID- 8220563 TI - [The story of Herman]. PMID- 8220561 TI - [Discussion about foreign nurses. Lack of information and follow-up]. PMID- 8220564 TI - [What really is nursing?]. PMID- 8220565 TI - [A glimpse of nursing--2 vignettes of patients who helped me to insight]. PMID- 8220566 TI - [Leadership of nursing. Development of professional nursing practice]. PMID- 8220567 TI - [Attitude to drug abuse among nursing students]. PMID- 8220568 TI - [AIDS in the world--WHO recommends prison reform]. PMID- 8220569 TI - [Rheumatic diseases. A national illness with an uncertain cause]. PMID- 8220570 TI - [Rheumatoid arthritis]. PMID- 8220571 TI - The effects of hormones on the gametocytogenesis of Plasmodium falciparum in vitro. AB - To examine possible effects of hormones on the gametocytogenesis of Plasmodium falciparum in vitro, insulin, progesterone, 17-beta-estradiol and testosterone were added in different concentrations to the culture medium. The medium containing RPMI was free of human serum but complemented with BMS (Basal-Medium Supplement, Biochrom), a semidefined serum replacement. As controls, cultures were grown either with BMS supplemented RPMI without hormones, or with human serum complemented RPMI. Only addition of insulin increased the parasitemia. All tested steroid hormones added to the medium enhanced the number of gametocytes. PMID- 8220572 TI - A survey of intestinal helminths among students of post-primary institution in Adamawa State, Nigeria. AB - A survey was carried out to determine the prevalence of intestinal helminthiasis among students of Government Secondary School Girei, Song Local Government Area of Adamawa State, Nigeria, between January and July 1991. Of the 200 students between ages ten to thirty-two years old examined, 86 (43.0%) were infected using the formyl ether concentration method. The most commonly found worm were hookworms, Ascaris lumbricoides and Trichuris trichiura with mean egg per gram of 4,800; 2,600; and 1,250 respectively. Infection was independent of both sexes, and parental occupations (P > 0.05) but decreased significantly with host age and a progressive increase in body weight (P < 0.05). Suggestions are made for the control of helminth parasites in the area. PMID- 8220573 TI - On the pathology and localisation of Cysticercus ovis in the skin of sheep. AB - Histological studies were conducted with experimentally and spontaneously infected lambs on the localisation of T. ovis larvae in the subcutaneous connective tissue and in the skin. In the first few days following infection an inflammatory process in the papillar and reticular skin layers was found, causing skin defects which appeared on tanning. Forty days after infection the mature cysticerci caused atrophic changes. Openings of 6-8 mm in diameter, evident on the tanning of the skin, appeared. These injuries reduced significantly the quality and the economic value of the skins. PMID- 8220574 TI - Falciparum malaria in naturally infected human patients: ultrastructural alterations of non-parasitized red blood cells during anaemia. AB - Several ultrastructural alterations have been displayed by non-parasitized erythroblasts (Eb) and erythrocytes (Ec) in the peripheral blood of 28 Saudi patients who were anaemic and suffering from acute falciparum malaria (AFM). A counting system is incorporated to determine the rate of altered to normal red cells in plastic serial sections. The Eb showed for the first time conspicuous surface knobs which were previously described only for parasitized Ec, and are known to play a role in their adhesion to vascular endothelial cells. The Ec displayed a bizarre contour and ragged appearance produced by the abundance of cytoplasmic processes. Such cells were similar to erythrocytic ropalocytes seen in haematological diseases associated with anaemia. The Ec also possessed autolysosome-like vacuoles containing rosette forms of alpha-glycogen-like particles (GLP) while smaller beta-GLP were dispersed in the cytoplasm. Ultrastructural alterations of these cells were suggested to be due to dyserythropoiesis and ineffective erythropoiesis, which were in turn attributed to an unbalance in metabolism as they were being overproduced in response to infection. Numerous haemoglobin-like particles (Hg) were being liberated through the erythrocytes plasma membranes indicating severe haemolysis which is considered to be one of the major factors in producing anaemia during malarial infection. This study showed tat anaemia is common in patients with AFM which is usually associated with structural alterations of non-parasitized immature and mature red blood cells (RBC). In addition, the rate of abnormal RBC correlates with the degree of parasitaemia. Moreover, old patients showed a lower degree of parasitaemia than young patients, possibly due to a less mature immune system in the later.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8220575 TI - [The parasitization of Gadus morhua in the southern Baltic Sea]. AB - During a survey of the working group 25 of BMB in the south of the Baltic Sea in October 1991 105 cod (Gadus morhua L.) were investigated for parasites (ICES Subdivision Areas 22, 24, 25 and 26). They were parasitized by 15 species. With increasing salinity (from east to west) and in the higher size groups more parasite species were present. Eimeria spec. in the liver and Schistocephalus spec. in the body cavity were registered for the first time in cod. PMID- 8220576 TI - [The effect of praziquantel on Trichobilharzia (Digenea, Schistosomatidae), a cause of swimmer's dermatitis in humans]. AB - Cases of swimmer's itch caused by cercariae of the genus Trichobilharzia have spread during the last years in Southwestern Germany and increased in intensity. The only consequence was to forbid swimming because no effective strategies are known which would be ecologically or financially acceptable. In this examination we therefore tried, by means of medication of ducks, the possible final hosts, to kill the different stages of Trichobilharzia in patency and prepatency in order to interrupt the parasitic cycle. For this the effectiveness of praziquantel on the parasitic stages in the final host (preadults, adults, eggs or fully developed miracidia) was tested in experimentally infected dwarf mallards (Anas platyrhynchos, hemerot.) and mallards (Anas platyrhynchos platyrhynchos). During therapy of ducks with patent Trichobilharzia infection, onefold or twofold application of this drug (1 x 500 mg/duck, 1 x 300 mg/duck, 2 x 200 mg/duck) only prevented the releasing of eggs with hatchable miracidia for up to 24 h. Only a threefold application of 200 mg/duck in 24 h intervals led to a permanent reduction of detectable miracidia. Application of praziquantel in low doses (30 or 40 mg per duck and day) did not reduce the number of released miracidia. Medication with praziquantel led to a strong shift of adult worms located in the enteric veins of the ducks to the liver veins in a little as 3 h; examinations by scanning electron microscopy revealed changes in the worms' surface and damage of the tegument. During prepatency doses of 22.5 mg praziquantel per duck and day, given continuously for one week, were sufficient to completely stop the release of miracidia. In this low dosage, a special preparation of the bitter drug was voluntarily swallowed by the ducks, thus providing in principal a simple, cheap and ecologically sound method of controlling cercarial dermatitis. PMID- 8220577 TI - [Faunistic-ecological studies of Ixodes ricinus in the urban area of Leipzig]. AB - The urban-ecological rule of decrease of the population size from the environs to the town centre is established also for Ixodes ricinus. The manifold impairments are shown which urban influences have on the cycle of the parasite, especially in the transitional zone between the forest area and the totally sealed ground. The presentation of an urban gradient is tried for the first time. There is potential danger of infection with Borrelia burgdorferi deep into the conurbation. PMID- 8220578 TI - Malaria morbidity among children attending maternal and child welfare clinics in northern Nigeria. AB - Blood samples from 522 children within the ages of 0-9 years attending maternal and child welfare clinics in northern Nigeria who were clinically diagnosed as being malarious were obtained for parasitological examination. Results revealed a parasite rate of 41%, density index of 7.72 and 19.5% of the population with parasite densities higher than the critical value of 10,000 parasites/mm3. There was no direct relationship between body temperature and rate of infection and between febrile illness and parasite density. Sex unlike age had no effect on the infection in the population. The study confirms that accurate diagnosis of malaria rests upon parasitological examination and not clinical acumen. PMID- 8220579 TI - Laboratory investigations on the larval zoophagy of Ophyra capensis--an antagonist of Musca domestica. AB - Second and third instar larvae of Ophyra capensis are distinct predators. Although facultative, their zoophagy is so strong that they show cannibalism in the absence of prey. If house fly larvae are available, the predators can kill up to 17 prey per larva of the predator depending on the larval instar and the culture density. The highest effectiveness under laboratory conditions was observed at densities from 1.7 to 3.0 g substrate per larva from the culture and at a ratio between predators and prey from 1:6 to 1:12. Under these conditions the reduction of house fly numbers as a result of O. capensis action is over 70%. If the natural elimination during house fly development (in our experiments from the second larval instar up to the adult eclosion it is not less than 10%) also is taken into account, it seems that under these conditions at the most 20% of M. domestica can develop to the adult stage. The similar requirements of biotopes and substrates for the preadult development of O. capensis and M. domestica, the high predator activity and the big food portion give a good reason to conclude that O. capensis is a perspective agent for biological house fly control. PMID- 8220580 TI - Similar simian virus 40-induced immortalization frequency of fibroblasts and epithelial cells from human large airways. AB - Several previous studies have shown that when Simian virus 40 (SV40) genes were introduced into normal human fibroblasts, a very small proportion (< 2%) of the resulting transformed clones were immortalization-competent. In separate studies with normal human epithelial cells, SV40 genes were shown to induce a much higher proportion (30-100%) of immortalization-competent clones. In this study, a direct comparison was made of the immortalization frequencies of clones of normal epithelial cells and fibroblasts derived from the same individual. The results demonstrate that fibroblasts and epithelial cells can be immortalized at similarly high frequencies when transfected with an SV40 early region origin minus expression plasmid, without the need for a selectable marker. We conclude that there is no intrinsic difference between the SV40 early region-induced immortalization potentials of human fibroblasts and epithelial cells. Comparison of this study with other studies of the immortalization of human fibroblasts suggests that removal of SV40 late region genes from the transfected plasmid is important for obtaining a high proportion of immortalization-competent clones. PMID- 8220581 TI - c-MYC mRNA is present in human sperm cells. AB - Stage-specific expression of proto-oncogenes, including c-myc, has been demonstrated during spermatogenesis in testis. Some of these proto-oncogenes are expressed postmeiotically, especially in the round spermatid stage. Recently, we demonstrated the presence of c-myc protein in mature ejaculated sperm cells with a possible role in sperm cell function. Since the half-life of c-myc protein has been shown to be short, we suspected the presence of c-myc mRNA in human sperm cells. In the present study, the presence of the c-myc mRNA transcript in human sperm cells was investigated by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and in situ hybridization. Total RNA, 5-10 micrograms, was extracted from 0.2-0.5 ml of pelleted human sperm cells by NP-40 lysis procedure, and was used to construct cDNA with pd(N)6 random primer and Moloney Murine Leukemia Virus (MMLV) reverse transcriptase. The PCR with sperm cDNA and primers #P1 and #P2, both from exon 3, resulted in amplification of the expected 322 bp product. Primers #P3 and #P4, which are located in exon 2 and exon 3, respectively, and are 1.37 kb apart, gave the expected PCR amplified 313 bp product ruling out the possibility of DNA contamination. The presence of c-myc mRNA in human sperm cells was further confirmed by in situ hybridization using a digoxigenin labelled DNA probe, containing exon 2 of the c-myc gene sequence. The c-myc specific DNA probe reacted with the postacrosomal mid-piece and tail regions of both noncapacitated as well as capacitated methanol-fixed sperm cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8220582 TI - MyoD transactivates angiotensinogen promoter in fibroblast C3H10T1/2 cells. AB - MyoD is one of the regulatory genes that causes activation of the expression of muscle specific genes in fibroblasts through interaction with a cis-acting DNA element CANNTG (E-box). We show here that forced expression of MyoD in mouse fibroblast C3H10T1/2 cells induces the expression of the liver specific rat angiotensinogen gene promoter. In the absence of MyoD, the angiotensinogen promoter is not expressed in C3H10T1/2 fibroblasts. Cotransfection of MyoD has no significant effect on the angiotensinogen promoter activity in the liver-derived HepG2 cells, suggesting that the regulatory programs of HepG2 and C3H10T1/2 cells are different, at least in terms of the requirement for MyoD-mediated transactivation of the target angiotensinogen promoter. PMID- 8220583 TI - H-DNA can act as a transcriptional insulator. AB - The polypurine/polypyrimidine DNA stretches with repetitive sequences (H palindromes) can adopt triplex-mediated folded structures called H-DNA. H palindromes are more represented than expected for a random distribution of bases in eucaryotic genomes, suggesting that they could ensure some biological function. Most studies have focused attention on their possible involvement in the control of transcription because of their particularly high frequency in the 5'-flanking sequences of genes. Using the (5'-TTCCC-3')n sequence present in the upstream region of several genes, this work concludes to a novel potentiality of H palindromes: The strong ability to disrupt the cooperation between proximal elements initiating transcription and distal elements enhancing transcription over a long distance. We present three structural features of the TTCCC repeat likely to explain such an activity: (1) the visualization by electron microscopy showing that a long H palindrome spontaneously forms higher order tertiary structures; (2) the fact that this structure is a target for specific nuclear proteins displaying an affinity for single-stranded polypyrimidines; and (3) the preferential localization of the genomic TTCCC repetitive sequences at the level of chromosomal matrix attachment regions (MARs). We propose that H-DNA can insulate some genetic loci from influences of their chromosomal environment, and belongs to a subclass of genomic matrix attachment regions. PMID- 8220584 TI - The novel purification of functional RNA polymerase III VAI transcription preinitiation complexes. AB - A novel purification scheme employing high-performance liquid chromatography (HPLC) gel filtration chromatography on a 4000 A methacrylate-based polymer and preparative gel mobility shift fractionation on 2% agarose gels with continuous elution allowed for a 10(5) fold enrichment in RNA polymerase III transcription preinitiation complexes from NTP-depleted HeLa whole cell extracts. Purified preinitiation complexes from the TATA-less Class III (Internal Control Region) adenovirus VAI gene contain the TATA-Box binding protein (TBP) by the criterion of comigration on sodium dodecyl sulfate (SDS) gels with the 43 kd TBP present in a purified B-TFIID fraction, and by Western blot analysis. Purified VAI preinitiation complex preparations lacking RNA polymerase III consistently contain a number of additional polypeptides with apparent molecular weights of 17, 20, 29, 32, 39, 55, 105, 120, and 140 kDa which appear to be specific components, since they are not detected in the corresponding fractions isolated using a VAI gene B-block mutant promoter. PMID- 8220587 TI - Bibliography of cellular and molecular biology research. PMID- 8220585 TI - The calcium binding protein of Entamoeba histolytica: expression in Escherichia coli and immunochemical characterization. AB - The gene encoding calcium binding protein (CaBP) of Entamoeba histolytica is single-copy and transcribes a single class of mRNA. The coding region of the gene, amplified from a genomic clone of CaBP by polymerase chain reaction, was cloned into the Escherichia coli expression vector pET-3c. The expressed protein was 15 kD in size, which agreed with the predicted size from the open reading frame. The recombinant protein showed altered mobility in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) after binding calcium and was found to be resistant to boiling, a property shared by many calcium binding proteins. A polyclonal antibody raised against the recombinant CaBP recognized a 15 kD protein in lysates from E. histolytica pathogenic strain HM-1:IMSS. The amebic protein could compete with the recombinant protein in binding to the antibody. While the antibody recognized lysates from a number of E. histolytica strains, it failed to cross-react with other Entamoeba species, e.g., E. invadens and E. moshkovskii. Moreover, no homologue of the CaBP gene could be detected in these species by using a set of degenerate primers for amplification by polymerase chain reaction (PCR). Our data demonstrate that amebic CaBP can be expressed in E. coli and that this protein is detectable only in E. histolytica but not in other Entamoeba. PMID- 8220586 TI - Restriction selection cloning: a simple general method for the selection of recombinant DNA. AB - It is well known that the ligation of two DNA fragments which are the product of digestion from different restriction enzymes will not lead to the regeneration of either of the original blunt end or cohesive end restriction sites. This property of sequence incompatibility for the original restriction enzyme can be exploited in a general cloning procedure for both PCR products and restricted DNAs. Restriction selection is particularly useful when cloning low abundance polymerase chain reaction (PCR) products and when cloning blunt ended DNA into reporter vectors that lack a method for the selection of recombinants. PMID- 8220588 TI - Acidic domain of the phosphoprotein (P) of vesicular stomatitis virus differentially interacts with homologous and heterologous nucleocapsid protein (N). AB - The homologous and heterologous interactions between the nucleocapsid protein N and the phosphoprotein P of New Jersey and Indiana serotypes of vesicular stomatitis virus were studied. SP6 derived N and P mRNAs were cotranslated in rabbit reticulocyte lysate and the complexes formed thereof were analyzed by 7.5% nondenaturing polyacrylamide gel electrophoresis. P protein of VSV(NJ) has two binding sites for homologous N protein: One located within the C-terminal 11 amino acids (within domain III) is responsible for the formation of five specific complexes while the other site, which spans the acidic domain I, is necessary for the formation of the sixth complex only. In contrast, P(IND) does not form the sixth complex when interacted with homologous N protein. Interestingly, P(NJ) forms only complexes 1 to 5 when it interacts with N(IND). The above results suggest that the complex 6 formation or domain I interacting site is NJ-serotype specific. Two chimeric P proteins were made using heterologous domains I and II/III of the P proteins of both serotypes. The soluble interaction of the chimeric proteins with the N protein supported the observed serotype specific interactions. The chimeric P proteins bound with equal efficiency with N-RNA template of both serotypes. These results strongly suggest that the acidic domain I of the P protein differentially interacts with homologous and heterologous N proteins. The biological significance of these findings is discussed. PMID- 8220589 TI - Pioneers' progress. Three states move ahead with their reforms. PMID- 8220590 TI - Full steam ahead! Survey confirms rush to join networks. PMID- 8220591 TI - Structure follows strategy. AB - Establishing the framework for an integrated health network is just one step in a complex process. Those building networks soon realize the skills requirements for network senior management differ from those for senior management in freestanding hospitals. So who will be on the new network management team? Experts say the answer depends on a variety of factors. PMID- 8220592 TI - Specialty gamble. Joining a health network is a high-stakes decision. PMID- 8220593 TI - Rough ride. EDs and trauma centers are buffeted by integration and managed care. PMID- 8220595 TI - Survey finds strong support for shared health benefits costs. PMID- 8220594 TI - Columbia, SC. Four area hospitals plant the seeds for closer ties, community planning. PMID- 8220596 TI - Clinton reform plan: health care vendors discuss implications. PMID- 8220597 TI - Sicker patients? New system helps evaluate proper utilization. PMID- 8220598 TI - 'Revolutionary' aspect of regulations gives HMOs more latitude. PMID- 8220599 TI - Six reasons why price controls won't work. PMID- 8220600 TI - Do computerized patient records risk invading patient privacy more than paper records? No. PMID- 8220601 TI - Do computerized patient records risk invading patient privacy more than paper records? Yes. PMID- 8220602 TI - At the grass roots. How I helped my community restore its health care system. Interview by Rhonda Bergman. AB - The late 1980s saw the unraveling of the health care system in rural northern Pend Oreille County, WA. It began when the area's only physician left, followed by the shuttering of the only area hospital. Kathy Grass, office manager of the local health clinic and a longtime hospital employee, was one of many who stepped forward to help. Given the title "community encourager" by other local volunteers, Grass began working with other residents to restore the local health care system and recruit providers. In a recent interview with Hospitals & Health Networks staff editor Rhonda Bergman, Grass described the challenges faced by her community. PMID- 8220603 TI - Getting down to details. Hospital groups take a careful look at the Clinton bill. PMID- 8220604 TI - Family practice physician survey. PMID- 8220605 TI - Learning by merging. Once adversaries, payers and providers develop new partnerships. AB - The rapid movement in health care toward the integration of delivery and payment systems reflects both a quickly changing marketplace and the anticipation of reform. The challenge in merging insurance and provider organizations, say those insurers who are doing it, is in the details. PMID- 8220606 TI - Learning by doing. Hospitals and physician groups move forward with insurance products. PMID- 8220607 TI - No more bells and whistles. Health care reform and network pressures are taking imaging back to basics. PMID- 8220608 TI - Opening arguments. Organized medicine warily eyes the Clinton plan. PMID- 8220609 TI - Lending a hand. Foundations see chance to speed up, complement reform. PMID- 8220610 TI - St. Louis providers shape market in advance of reform's tide. PMID- 8220612 TI - Local alliance calms tempers and tackles health costs. PMID- 8220611 TI - Data watch. Study: hospitals intensify their recruitment of primary care physicians. PMID- 8220613 TI - Rule 1 on critical paths: proceed with caution. PMID- 8220614 TI - The immunization imperative. PMID- 8220615 TI - The substance of names. PMID- 8220616 TI - Analyzing consumer magazine markets. AB - This nurse author has experience writing for the public. She shares her tips on how to analyze the consumer magazines in order to develop a successful manuscript targeted to their readership and style. PMID- 8220617 TI - Review process for nursing journals: making the best of any review system. AB - Within nursing, different review practices exist in the operation of nursing's professional journals. This article describes each of the three major review practices and provides you with some tips on how to work with these approaches as an author or editor. PMID- 8220618 TI - Reviewing manuscripts: how to be critical without being offensive. AB - Authors often take reviewers' comments to heart. Develop your manuscript review skills so that you give positive criticism to the author without making personal or professional attacks on him or her. PMID- 8220619 TI - Nurse author's guide to publishing house editors. AB - When you write an article or book manuscript, you are likely to work with several editors, including the acquisitions editor, developmental editor, production editor, and others. By knowing what each editor is responsible for, you will be able to communicate with the right person when you have a question or need help on your manuscript. PMID- 8220620 TI - I sent a query letter to a nursing journal editor describing a potential article. PMID- 8220621 TI - Reviewing manuscripts: developing an efficient system. AB - By focusing on accuracy, organization, and clarity you can develop an efficient system for reviewing manuscripts. This experienced reviewer tells you what to focus on in your manuscript review. PMID- 8220622 TI - Elements of an effective query letter. PMID- 8220623 TI - Writing transition sentences. AB - Transition sentences help the reader move from point to point. They clarify a key point, so the reader can distinguish main points from specific details in an article or book. In this article, you will be able to practice writing transition sentences. PMID- 8220624 TI - Case management studied. PMID- 8220625 TI - Health changes cause problems. PMID- 8220626 TI - Sexuality and cancer. PMID- 8220627 TI - Nurses' unity forces changes. PMID- 8220628 TI - Pulse oximetry: shedding a little light. PMID- 8220629 TI - Developing professionalism. PMID- 8220630 TI - New approaches to technology assessment: opportunities and trends. PMID- 8220631 TI - The clinical engineer's role in incident investigation. PMID- 8220632 TI - Telemetry for the year 2000: meeting the needs of the customer. AB - As customers of medical technology, all too often we are limited in our equipment selection process by the unwillingness of the vendor to custom-design their equipment to meet our needs. Reductions in capital equipment budgets as well as in operating (salary) budgets will require hospitals to make equipment selections based not only on how the technology can fulfill a patient care need, but also on how the technology can reduce or offset operating costs. The replacement of older telemetry monitoring equipment with state-of-the-art computerized, distributed network display terminals offers a new insight into how telemetry monitoring can better serve the needs of the caregiver, and at the same time contribute to reductions to operating costs. Computerized arrhythmia telemetry monitoring technology has finally advanced to the point where you don't need a full-time person to "monitor the monitor." PMID- 8220633 TI - EquipTeach: a computer-aided instruction to teach users how to operate specific medical equipment. PMID- 8220634 TI - Avoiding tourniquet-induced neuropathy through cuff design. AB - Pneumatic tourniquets are routinely used in limb surgery to provide a bloodless operating field, but they are known to cause nerve injuries. Simulation results based on the hypothesis that axial compression of nerves is responsible for a certain class of these mechanically induced injuries are in substantial accord with clinical observations. A model of the limb and tourniquet that treats the tissue as a linearly elastic solid is presented and is used to predict the induced axial strains. The smallest axial strains are induced by a tourniquet design for which the applied pressure distribution rolls off as gradually as possible; according to the axial-strain hypothesis, such a design will markedly decrease a tourniquet's inherent potential for injury. Use of a wider cuff in and of itself will not reduce axial strain, so if the hypothesis is correct, a wider cuff would not be intrinsically safer than a regular cuff, a result that is contrary to current opinion. PMID- 8220635 TI - Neural-network applications for analysis of adverse drug reactions. AB - A neural-network analysis has been applied to predict adverse or side effects of drugs using a database of antidepressant agents with known effects as well as input from a database with both patient parameters and drug information. A NeuralWork software package was implemented on a Macintosh Quadra 700 and trained on a database of ten drugs with known adverse effects. Another agent (not in the database) was used to test the ability of the network to predict the relative incidence of its side effects. Despite the small number of drugs used for training, the adverse effects of some drugs, such as doxepin, were predicted with 90-100% accuracy. These results indicate that neural-network analysis can be used to predict adverse drug effects for drugs within a given class and ultimately can be extended to include patient parameters to predict the mechanisms of action of drugs from relatively large databases. PMID- 8220636 TI - Steam sterilization--the response of the test pack. AB - The authors obtained temperature recordings from nearly 400 experiments with the standard test pack as defined in the prEN 285 as a basis for methods to evaluate conditions influencing steam sterilization. A new method to scale the duration and the extent of an air pocket (penetration fault) is presented. This method proved suitable for the interpretation and quantification of substantial correlations between the effect of a fault, the volume of the sterilizer chamber, and the rate of pressure change in the chamber. The coolest spot was found near the center of the test pack in only 17% of the tests. The air pocket was found to be an anomaly best elucidated by statistical methods. The prevailing influence of the rate of pressure change does not allow the establishment of limiting values for any fault, such as leakage rate or air dilution in the chamber, except as applied to a well-defined sterilizer. The penetration fault caused by an air leak does not depend on the chamber volume. The effects of residual air and induced air on the penetration fault are fundamentally different. Faults caused by those different air sources should be distinguished from each other even though both cause air pockets. PMID- 8220637 TI - India--critical care help for Mother Teresa. PMID- 8220638 TI - Assessing and treating decreased cardiac output. AB - The compensatory pathophysiological mechanisms for a decreased cardiac output are explored via three case studies. The assessments and nursing interventions appropriate to these patients are included in separate tables. Evaluation and decision making about the patient's clinical status and responses are major nursing responsibilities that require astute assessment and skilled intervention to lead to optimal patient outcomes. PMID- 8220639 TI - Postoperative management of patients undergoing radical cystectomy and urinary diversion. AB - The management of patients undergoing radical cystectomy and urinary diversion requires an understanding of the various tubes and drains used in reconstructing the urinary tract. The three major types of urinary diversions are presented with particular attention to the nursing skills required for successful postoperative care. PMID- 8220640 TI - Systemic lupus erythematosus. Part 2: Diagnosis, treatment modalities, and nursing management. AB - Recent advances in the diagnosis and symptom management of systemic lupus erythematosus have now made the disease compatible with normal life span. However, systemic lupus erythematosus remains a multisystem disorder with no cure. Consequently, every specialty, particularly medical-surgical nursing, is challenged to provide comprehensive care to these patients. PMID- 8220641 TI - Reducing occupational transmission of tuberculosis to health care workers. AB - Tuberculosis infection is an occupational risk for nurses and others working in health care settings. The National Institute of Occupational Safety and Health (NIOSH) and the Centers for Disease Control (CDC) have issued guidelines for preventing transmission of tuberculosis to health care workers. Nurses should be familiar with these guidelines, current recommendations for protection and post exposure management, and the implications of the resurgence of tuberculosis as an occupational health hazard. PMID- 8220642 TI - Factors contributing to protein-calorie malnutrition in older adults. AB - Protein-calorie malnutrition, a common problem of older adults, is frequently overlooked by nurses. The physiologic changes of aging, as well as changes in living conditions and income, all contribute to the problem. Recognition of this problem by nursing staff can reduce many of the health problems which are exacerbated by or result from malnutrition in this patient population. PMID- 8220643 TI - Development of a nursing information systems department. Part 2: Operationalizing the targets, strategies, and tactics. PMID- 8220644 TI - Preventing malpractice. PMID- 8220645 TI - What to expect from an ethics consultation. PMID- 8220646 TI - Unit-based educators: decentralizing resources at the unit level. PMID- 8220647 TI - The art of setting limits. PMID- 8220648 TI - Pharmacologic management of systemic lupus erythematosus: nursing implications. PMID- 8220649 TI - Planning to meet the JCAHO standard: consideration of scientific findings. AB - Bringing research findings into practice does take some thought and planning, but most health care institutions already have committees, departments, and positions that can facilitate research utilization. It may just take some assessment of the roles and functions of these components to more carefully plan for systematic use of research findings in practice. This planning will pay off by enhancing the quality of nursing care and providing documentation that review of policies and procedures includes consideration of current scientific and nursing research findings. PMID- 8220650 TI - Haemorheological changes in obese adolescents after short-term diet. AB - Along with other risk factors in coronary heart and cerebrovascular disease, obesity is frequently associated with the development of atherosclerosis, a disease in which the rheological characteristics of blood are important. We studied the influence of weight reduction on haemorheological parameters of 20 obese adolescents (10 female, 10 male; aged 12-17 years) without metabolic and/or cardiovascular diseases. The control group composed 39 health non-obese subjects (19 female, 20 male; aged 13-17 years). At the beginning of the study the mean (+/- s.d.) body mass indices (BMI) of the obese adolescents were 36.0 +/- 5.3 kg/m2 in males and 36.8 +/- 5.0 kg/m2 in females. This was significantly reduced (31.5 +/- 4.9 kg/m2 in males and 32.5 +/- 4.7 kg/m2 in females) after one month following a diet of 1000 kcal/day: 25% proteins, 26% lipids and 49% carbohydrates. Before dieting, plasma fibrinogen level, whole blood viscosity at low shear rates and plasma viscosity were significantly higher in obese patients than in normal subjects, while the microhaematocrit values of the two groups were not significantly different. After dieting, plasma viscosity, the plasma fibrinogen level and the mean erythrocyte aggregation index of the patients decreased significantly compared to basal values, while whole blood viscosity did not change. Our data seem to indicate that short-term diet improves the haemorheological pattern in obese adolescents, probably as a consequence of an important metabolic rearrangement. PMID- 8220651 TI - Long-term effects of feeding high carbohydrate diet in pre-weaning period by gastrostomy: a new rat model for obesity. AB - In the present study, we report the long-term metabolic consequences of feeding a milk substitute formula that is high in carbohydrate-derived calories during the suckling period. Male Sprague-Dawley rat pups were raised by gastrostomy on a high carbohydrate (HC) formula or a high fat (HF) formula (which mimicked rat milk composition in macronutrients) during the pre-weaning period (day 4 to 24). These rats were then weaned to a laboratory stock diet and subsequently challenged with a high sucrose diet to augment the development of obesity. The pups receiving the HC formula developed obesity in later life, even though there was no change in the body weight of this group compared to mother-fed (MF) controls or HF formula fed animals during the pre-weaning period. The HC rats were hyperinsulinemic and their growth rates were greater than MF rats starting at day 55. The lipogenic capacity of liver as well as adipose tissues (epididymal and omental) was higher in HC animals compared to MF control animals, as reflected by increases in two key lipogenic enzymes (fatty acid synthase and glucose-6-phosphate dehydrogenase) and in vitro synthesis of lipids. An analysis of adipose tissue morphology in adult rats showed an increase in cell size in epididymal adipose tissue of HC rats compared to the MF group. However, there was no significant difference in cell size in omental adipose tissue between the MF and HC rats. The HF group behaved similarly to the MF control group in growth pattern and measured metabolic parameters. PMID- 8220652 TI - Swedish obese subjects (SOS)--an intervention study of obesity. Baseline evaluation of health and psychosocial functioning in the first 1743 subjects examined. AB - This part of an on-going intervention trial analyses impacts of obesity on psychosocial factors and health. The study sample comprised 800 obese men (BMI > or = 34 kg/m2) and 943 women (BMI > or = 38 kg/m2) ranging in age from 37 to 57 years. All participants completed standardized health-related quality of life measures, a validated obesity-specific eating inventory and study-specific questionnaires on current and past health status, use of medical care and medications, socioeconomic status, dietary habits, physical activity habits, weight history and familial history of obesity. Chronic patients and population samples were used as reference. The obese reported distinctly poorer current health and less positive mood states than the reference subjects, women being worse than men. Anxiety and/or depression on a level indicating psychiatric morbidity were more often seen in the obese and again women reported more affliction than men. Furthermore, the average poor mental well-being was worst than in chronically ill or injured patients, such as rheumatoid, cancer survivors and spinal cord injured persons. Predictors of perceived health and psychosocial functioning could be discerned using a comprehensive system of statistical analyses (16-28% explained variance). A background of both somatic and psychiatric morbidity was decisive for the health and psychosocial functioning in the obese; joint symptoms and angina pectoris dominated among somatic variables. Physical inactivity was the most prominent of traditional risk factors. The number of dieting attempts and body image were important weight correlates. Our results provide further evidence to the effect that severe obesity is a crippling condition. PMID- 8220654 TI - Circadian rhythms of appetite at different stages of a weight loss programme. AB - A group of 28 healthy obese people (16 women, 12 men) rated their appetite each whole hour of the day during nine separate days at three different stages of a dietary intervention study of 24 weeks. The first stage was an introduction period of eight weeks in which subjects were weight stable. The second stage was a weight reduction period of 13 weeks in which subjects lost about 0.85 kg/week of body weight. In the third stage of three weeks subjects were weight stable again. The slimming diet, which contained predominantly sweet items, had an energy content which was 1000 kcal (4.2 MJ) below the individually estimated daily energy expenditure. The results showed that appetite for a meal increased in the first week of the weight reduction period of 13 weeks but returned to the baseline level within 4-8 weeks. Appetite for something savoury remained elevated during the entire weight reduction period, and returned to the baseline level in the stabilization period. Appetite for something sweet did not change. The circadian rhythms of the different types of appetite remained unchanged at all stages during the diet intervention programme. Different types of appetite (e.g. sweet, savoury) had different patterns over the day. PMID- 8220653 TI - Evidence for brain serotonin-mediated control of carbohydrate consumption in normal weight and obese humans. AB - A plasma insulin and amino acid-mediated mechanism is thought to modulate brain serotonin concentration, thereby regulating carbohydrate consumption on a meal to meal basis. It has been suggested that obesity is associated with a defect in the appetite control system. Furthermore, post-absorptive plasma levels of several amino acids are increased in obese subjects, which is ascribed to obesity associated insulin resistance and/or hyperinsulinemia. We studied breakfast induced changes in plasma ratios of tryptophan to other large neutral amino acids and associated differences in macro-nutrient composition of lunch food in normal weight and obese human subjects. The study was randomized, double blind and cross over with a 2 x 2 factorial design with drug/placebo and type of breakfast as factors. Nineteen healthy, non-obese (body mass index (BMI) 22.5 +/- 1.9 kg/m2, mean +/- s.d.) and 19 obese (BMI 34.7 +/- 6.2 kg/m2) female volunteers were treated with either 60 mg fluoxetine (FXT), a serotonin re-uptake blocker specifically acting in the brain, or placebo for four days with a wash-out period between treatments of four weeks. The subjects received either a carbohydrate (CHO) breakfast (80 g maltodextrin, 300 kcal) or a protein-rich (PROT) breakfast (60% milk protein and 40% CHO, 300 kcal) on two consecutive days (days 4 and 5 of each treatment period). Plasma glucose, insulin and amino acids were measured at several time points after breakfast. Three hours after breakfast, subjects were able to choose from 29 different food items. Total energy content and weight of lunch food and energy percentage of each macronutrient were calculated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8220655 TI - Measurement of excessive appetite and metabolic changes in Prader-Willi syndrome. AB - The behavioural, cognitive and metabolic response to food intake was studied in 13 adults with the Prader-Willi syndrome (PWS) and compared to ten age-matched controls. Rates of eating were observed during one hour's access to food and feelings of hunger were assessed using a visual analogue scale. Blood was taken for estimation of glucose, insulin, cholecystokinin (CCK), prolactin, growth hormone (GH) and cortisol every 20 min for a total period of 100 min. Ten (76%) of the subjects with PWS ate steadily for the whole hour that food was available and on average consumed three times more calories than the control group. The median ratings for feelings of hunger in the PWS group changed in the expected direction but these changes were delayed compared to the control group and only reached the same level as the controls after the PWS subjects had eaten a significantly greater amount of food. In the PWS group, in contrast to the control group, feelings of hunger started to re-emerge shortly after food was removed. There were marked differences between individuals with PWS in the extent of the changes in serum prolactin levels. Increases in plasma glucose levels were inversely correlated with changes in hunger ratings in the PWS group, but not the control group. There was a significantly greater increase in serum CCK levels during the meal in the PWS group than in the control group indicating that in PWS failure of peripheral release of CCK in response to food intake was not the explanation for the impaired satiety response. PMID- 8220656 TI - Snoring and sleep apnoea in men: association with central obesity and hypertension. AB - The objective of this study was to examine the relationship between sleep apnoea, measures of obesity and blood pressure. The study was cross-sectional in design, involving 1464 consecutive men undergoing sleep studies at two non-hospital sleep laboratories. Detailed sleep studies, body mass index (BMI), neck, hip and waist circumferences and morning and evening blood pressures were measured in each patient. Twenty-eight per cent of patients were obese (BMI > 30 kg/m2) and 47% were overweight (BMI 26-30 kg/m2). Body fat distribution in these patients was typically central with a mean waist-hip ratio (WHR) for the entire group of 0.99 with over 80% of patients having a WHR more than 0.94. Increasing severity of sleep apnoea measured either by respiratory disturbance index (RDI) or minimum oxygen saturation in sleep (MOS) was associated with increasing central obesity and morning but not evening blood pressure. Normal weight patients (BMI < 25 kg/m2) with sleep apnoea were characterized by increased waist and hip circumferences and increased morning diastolic blood pressure compared to patients without sleep apnoea. The best explanatory variables for sleep apnoea were waist (r2 = 0.156, P < 0.001) and age (r2 = 0.013, P = 0.003) only. Morning but not evening blood pressure was related to sleep apnoea independently of obesity and age. We concluded that patients with sleep apnoea are centrally obese. Waist circumference is a better predictor for sleep apnoea than neck circumference or BMI suggesting that the link between obesity and sleep apnoea cannot be explained solely by neck fat deposition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8220657 TI - Twenty years of cooperation between Polish and German pharmacologists. PMID- 8220658 TI - Open field behavior of rats reared in different social conditions: the effects of stress and imipramine. AB - Behavioral changes in rats divided on the weaning day into two groups (crowded and isolated) were studied in the computerized open field. At the end of the experiment the effects of an acute stress (1 h long immobilization) on open field behavior, body weight and body temperature, were examined. In a separate experiment both groups of rats were chronically pretreated with tricyclic antidepressant imipramine (5.0 mg/kg, po), and subsequently exposed to the same stress procedure. Social deprivation significantly enhanced spontaneous locomotion and exploratory activity. Restraint stress significantly decreased exploration, body weight and temperature, in the isolated animals only. Chronic pretreatment of rats with imipramine significantly attenuated the effect of stress on motor and exploratory behavior, as well as stress-induced changes in body weight and temperature, in the group of isolated animals. The present data indicate sensitization to stress of behavioral and vegetative processes in socially deprived animals. The relationship between rearing conditions and behavioral and vegetative reactivity to the environmental challenges is confirmed. PMID- 8220659 TI - A search for new active amino acids responsible for pharmacological activity of collagen degradation products. AB - The pharmacological activity of glycine (Gly), proline (Pro) and hydroxyproline (Hyp)--the main amino acids in collagen degradation products--was studied in comparison to the activity of all mixture of DPTC (degradation products of trihelical collagen). The investigated amino acids decreased psychomotor activity of rats, enhanced apomorphine- and amphetamine-induced stereotypy. Pro and Hyp enhanced also catalepsy, but Gly decreased the haloperidol action. Each amino acid was given adequately to its contents in DPTC. Our results indicated that hydroxyproline revealed the action on the central nervous system (CNS) in each aspect most similar to the action of DPTC mixture. PMID- 8220660 TI - The analgesic activity of some tuftsin- and tuftsin inhibitor-like fragments of the viral coat proteins. AB - Antinociceptive activity of seven pentapeptide fragments of human adenovirus type 2 (Ad2) virion proteins: Thr-Val-Pro-Pro-Arg (1), Thr-Arg-Pro-Pro-Arg (2), Thr Gly-Pro-Pro-Thr (3), Pro-Arg-Pro-Pro-Thr (4), Phe-Val-Pro-Pro-Arg (5), Ala-Arg Pro-Pro-Ala (6), Tyr-Gly-Pro-Pro-Lys (7)--analogs of known tuftsin inhibitor Thr Lys-Pro-Pro-Arg, was measured by hot-plate procedure. Also two tuftsin-like fragments of epitopes of HIV-1 and HIV-2: Thr-Lys-Ala-Lys (8), Thr-Lys-Glu-Lys (9), and tuftsin analog Thr-Lys-Asp-Lys (10) were tested. In the control experiments the effects of tuftsin and pentapeptide tuftsin inhibitor were also studied. The peptides 2, 4 and 5 were found to produce very strong analgesia after the icv injection. It was observed that pretreatment with peptide 8 remarkably diminished the antinociceptive effect induced by tuftsin. PMID- 8220661 TI - Hypothalamic, striatal and plasma leucine enkephalin level in two rat models of portal hypertension. AB - In two rat models of portal hypertension obtained by common bile duct obstruction or by partial portal vein ligation the hypothalamic level of leu-enkephalin was increased but striatal and plasma level of leu-enkephalin was unchanged. These results may be of importance for understanding some disturbance related to hepatic encephalopathy. PMID- 8220663 TI - Ambroxol inhibits endotoxin-induced lipid peroxidation in mice. AB - Administration of ambroxol (70 mg/kg ip) once a day for 3 days protected lung and heart lipids from lipopolysaccharide (LPS, 17 mg/kg)-induced oxidative stress in mice. Ambroxol as a lipid peroxidation inhibitor was almost as active as an equivalent dose of N-acetylcysteine (27.6 mg/kg), a well known antioxidant. The lung and heart levels of conjugated dienes in animals pretreated with ambroxol were 3.3- and 1.7-times lower (p < 0.05 and p < 0.01) than those observed in the control group which received only buffer and subsequently LPS. These results indicate that ambroxol can sufficiently inhibit the harmful process of lipid peroxidation in vivo. PMID- 8220662 TI - The responsiveness of M2-muscarinic receptors in the posterior hypothalamus and brain stem of vasopressin hypertensive rats. AB - The response of an endogenous inhibitor of cAMP-dependent protein kinase (type I inhibitor) to tremorine was used as an index of sensitivity of control muscarinic M2-receptors. Tremorine induced a dose-dependent increase in type I inhibitor activity in the posterior hypothalamus and brain stem. The action of the compound was blocked by pretreatment with aminophylline and atropine. Prolonged, 28 days treatment with lysine vasopressin (1 U/kg/day ip) induced hypertension and modified the dose-response curve for tremorine. Five times higher doses of tremorine than in normotensive rats were necessary to induce statistically significant increase in type I inhibitor activity in the posterior hypothalamus and brain stem suggesting subsensitivity of M2-muscarinic receptors in the brain areas responsible for the regulation of blood pressure. PMID- 8220665 TI - Critical care and healthcare reform. PMID- 8220664 TI - Similar action of imipramine and arginine-vasopressin in the social interaction test. AB - An antidepressant drug, imipramine, given chronically increased the social interaction between rats. A neuropeptide, vasopressin, demonstrated a similar, although weaker effect. PMID- 8220666 TI - Case studies in terminal weaning from mechanical ventilation. AB - Terminal weaning, withdrawal of mechanical ventilation when the patient is not expected to survive the process, must not be burdensome to the patient or significant others. The healthcare team must individualize the weaning process, considering the physiologic comfort of the patient and the psychoemotional comfort of both the patient and family. The following case studies illustrate variations in a method for terminal weaning that are patient-specific and are based on the experience of a supportive care team. PMID- 8220667 TI - Temporal patterns of heart rate and selected arrhythmias for 48 hours after cardiac surgery. AB - OBJECTIVE: To describe the temporal patterns of heart rate and arrhythmias in the immediate postoperative period following cardiac surgery. Six postoperative cardiac surgical patients with a mean age of 48.3 years were studied. DESIGN: Descriptive longitudinal design. SETTING: Cardiac surgical ICU. METHODS: Heart rate and arrhythmias were recorded continuously for 48 hours from a cardiac monitor using a Holter tape recorder. Environmental and treatment data were noted throughout data collection by trained nonparticipant observers. RESULTS: Mean heart rate and incidence of arrhythmias were different between the 2 study days; therefore, data were divided into two segments (A and B). These differences coincided with extubation in most cases. Individual subject cosinor analysis revealed 24-hour rhythms of heart rate in both segments in all subjects except segment B for one subject. Rhythms of shorter periods were also found. In segment A individual subjects' acrophases (peak times of fitted curves) occurred later than expected for subjects' prehospitalization sleep-wake schedule, whereas in segment B they occurred earlier. Cosinor analysis of arrhythmias revealed significant 24-hour rhythms in both segments in one of the three subjects with premature atrial complexes, two of the four subjects with premature ventricular complexes and both subjects with ventricular couplets. Four-hour rhythms were found in premature atrial complexes (n = 1), atrial tachycardia (n = 1) and premature ventricular complexes (n = 3). Acrophases for arrhythmias varied among patients. During segment B the 4-hour-rhythm acrophases in heart rate and arrhythmias were related to the timing of respiratory therapy. CONCLUSIONS: Temporal variations in heart rate could be identified in these six critically ill adults. Rhythm parameters changed during the first 48 hours after cardiac surgery. In those who had arrhythmias, some patients demonstrated temporal patterns in the incidence of selected arrhythmias. Further study is needed to describe the temporal patterns of heart rate and arrhythmias in varied groups of critically ill persons in a variety of settings. PMID- 8220668 TI - Management of delirium associated with use of the intra-aortic balloon pump. AB - Five patients who developed an agitated delirium in association with use of an intra-aortic balloon pump are presented. The differential diagnosis of delirium in critically ill cardiac patients is reviewed and effective pharmacologic treatment strategies (involving rapid and aggressive management with intravenous haloperidol) are discussed. PMID- 8220669 TI - Pediatric critical care nurses' knowledge of cardiac dysrhythmias. AB - OBJECTIVE: To describe pediatric critical care nurses' knowledge of dysrhythmias in critically ill pediatric patients and relate this knowledge level to certain demographic variables (education, nursing experience, certification, supplemental training, area of employment and geographic region of residence). DESIGN: A descriptive survey. SETTING: American Association of Critical-Care Nurses' 19 geographic regions of the United States. PARTICIPANTS: Of 1000 questionnaires mailed to pediatric critical care nurses who were members of the American Association of Critical-Care Nurses in 1991, 356 responses were received (a response rate of 36%). INTERVENTION: A criterion-referenced, self-administered test regarding pediatric dysrhythmias and a demographic sheet randomly mailed to 1000 pediatric critical care nurses. Test results were analyzed and compared with demographic variables. RESULTS: The mean total test score was 66%. Significantly higher total test scores and selected subtest scores were demonstrated in relationship to the following variables: increased age; certification in pediatric advanced life support, advanced cardiac life support or adult critical care; increased years of adult critical care experience; advanced dysrhythmia courses and dysrhythmia self-study; and perceived knowledge level above that of the advanced beginner. CONCLUSIONS: Pediatric critical care nurses' overall knowledge of dysrhythmias was low. Knowledge strengths included recognition of basic and life-threatening dysrhythmias and calculation of basic ECG measurements. Knowledge deficits included importance of sinus bradycardia in the neonate, appropriate intervention for life-threatening dysrhythmias and calculation of an irregular heart rate. These deficits should be considered when planning continuing education programs for pediatric critical care nurses. PMID- 8220670 TI - HELLP: a collaborative challenge for critical care and obstetric nurses. AB - Hemolysis, elevated liver enzymes and low platelet count, or HELLP syndrome, is a manifestation of pregnancy-induced hypertension. It is a multisystem, life threatening disorder. Pregnant women with HELLP syndrome often require critical care nursing. Collaboration between the critical care nurse and the obstetric nurse is essential because it enhances the nursing management of the mother and her fetus. This article reviews selected physiologic changes in pregnancy and the pathophysiology of pregnancy-induced hypertension and HELLP syndrome. These changes and pathophysiology serve as the rationale for the nursing care guidelines. PMID- 8220671 TI - Severe HELLP syndrome presenting with acute gum bleeding following toothbrushing at 38 weeks' gestation. AB - We describe a case of hemolysis, elevated liver enzymes and low platelet count (HELLP syndrome) at 38 weeks' gestation. This condition was diagnosed when the patient presented with acute, persistent gum bleeding after toothbrushing. Her platelet count was 26,000/mm3. Bleeding gums have rarely led to the diagnosis of this syndrome; this case appears to be the most severe description of this clinical presentation following toothbrushing yet presented in the literature. Although the patient had been hospitalized 48 hours earlier with nonspecific viral-like symptoms, she was discharged after being treated with intravenous hydration. Reexamination of laboratory studies obtained at the time of her first presentation revealed thrombocytopenia. The diagnosis of this syndrome could possibly have been made at that time. This case emphasizes the importance of maintaining a low threshold of suspicion for this syndrome. PMID- 8220672 TI - Sotalol hydrochloride (Betapace): a new antiarrhythmic drug. AB - Sotalol hydrochloride (Betapace), recently released by the Food and Drug Administration for general use, is used to treat a variety of ventricular and supraventricular tachyarrhythmias. The drug's dominant action is the result of combined nonselective beta-adrenergic antagonism (Class II effect) and monophasic action potential duration prolongation in all cardiac tissues (Class III effect). It causes less left ventricular depression than propranolol and has a low incidence of toxicity. It is a useful addition to the antiarrhythmic drug armamentarium. This article reviews the drug's pharmacokinetic, pharmacodynamic and electrophysiologic properties, clinical uses and potential side effects. Reports on the drug's use as an antianginal and antihypertensive agent are also discussed. PMID- 8220673 TI - Critiquing and using nursing research: guidelines for the critical care nurse. AB - This article focuses on a concern that practicing nurses are expected to use nursing research but are not thoroughly prepared to meet this expectation. This concern is addressed by proposing: (1) a guideline for research critique that is designed to bridge baccalaureate education and nursing practice and (2) a unique description of research utilization that may be realistically accomplished by practicing nurses. PMID- 8220674 TI - Reviews and summaries of research related to AACN 1980 research priorities: clinical topics. AB - This article reviews and summarizes the research conducted following publication of the 1980s American Association of Critical-Care Nurses' clinical research priorities. Original research conducted on the clinical priority topics between 1981 and 1991 was included. Review articles, doctoral dissertations, theses and abstracts were excluded unless judged to provide important information on the topic. Following the statement of each priority, progress in the area is summarized. Limitations and measurement issues are discussed as appropriate. Recommendations for future research are provided, and progress in the area is summarized. PMID- 8220675 TI - Case study: implications of the recommendation to the patient with an AICD to avoid magnetic fields. PMID- 8220676 TI - Syndrome X: fact or fancy? AB - Syndrome X continues to be a puzzle to the profession. A consensus as to the definition, etiology, pathology, prognosis and treatment has still to be developed. PMID- 8220677 TI - "Do not resuscitate' orders and end-of-life care planning. PMID- 8220678 TI - Open visiting policies in the ICU. PMID- 8220679 TI - Open visiting policies in the ICU. PMID- 8220680 TI - Open visiting policies in the ICU. PMID- 8220681 TI - Minimizing diagnostic blood loss in critically ill patients. PMID- 8220682 TI - Clinical practice guidelines. PMID- 8220683 TI - Guidelines for the transfer of critically ill patients. PMID- 8220684 TI - Probabilistic and decision-theoretic systems in medicine. PMID- 8220685 TI - Multiply sectioned Bayesian networks for neuromuscular diagnosis. AB - A prototype neuromuscular diagnostic system (PAINULIM) that diagnoses painful or impaired upper limbs has been developed based on Bayesian networks. This paper presents nonmathematically the major knowledge representation issues that arose in the development of PAINULIM. Motivated by the computational overhead of large application domains, and the desire to provide a user with an interface that gives a focused display of a subdomain of current interest, we built PAINULIM using the idea of multiply sectioned Bayesian networks. A preliminary evaluation of PAINULIM with 76 patients has demonstrated good clinical performance. PMID- 8220686 TI - A tutorial introduction to stochastic simulation algorithms for belief networks. AB - Belief networks combine probabilistic knowledge with explicit information about conditional independence assumptions. A belief network consists of a directed acyclic graph in which the nodes represent variables and the edges express relationships of conditional dependence. When information about one variable's state is given to the network in the form of evidence, an update algorithm computes the posterior marginal probability distributions for the remaining variables in the network. Many algorithms for performing this inference task have been proposed. Exact algorithms report precise results for some classes of networks, but take exponential time (in the number of nodes) both in the worst case and for many interesting networks. Stochastic simulation algorithms estimate the posterior marginal probability distribution for many graph topologies that would require exponential time when using an exact algorithm. Nonetheless, for some belief networks, stochastic simulation algorithms are also known to have exponential worst case performance. This article describes at a tutorial level several stochastic simulation algorithms for belief networks, and illustrates them on some simple examples. In addition, the theoretical and empirical performance of the algorithms is briefly surveyed. PMID- 8220687 TI - Computing the confidence in a medical decision obtained from an influence diagram. AB - Willard and Critchfield [31] assume that uncertainty exists as to the values of the probabilities which need to be assessed for a decision tree, and that the uncertainty in each probability is represented by a continuous probability distribution. If the probability itself is thought of as an objective limit of a relative frequency, then this distribution represents our degree of belief concerning the 'true' value of the probability. Willard and Critchfield [31] obtain a method which is able to determine the probability that the recommended decision is the one which would be obtained using the objective relative frequencies. This probability is called the confidence in the decision. These same results are obtained here for the case where a problem is represented in an influence diagram. There is also a discussion concerning the importance of the confidence measure in the evaluation of the quality of a medical expert system and in the instance of a single decision. PMID- 8220688 TI - Bayesian inference for model-based segmentation of computed radiographs of the hand. AB - We present a method for medical image understanding by computer that uses model based, hierarchical Bayesian inference to accurately segment imaged anatomy. A first application is a prototype system that automatically segments and measures symptoms of arthridities in hand radiographs. This is potentially useful in radiological diagnosis and tracking of arthridities. Key steps of the model based, Bayesian inference approach are: (1) prediction of imagery features from 3D models of anatomy, parameterized by population statistics, (2) local image feature extraction in predicted sub-regions, and (3) the use of a probabilistic calculus to accrue results of image processing and image feature matching procedures in support or denial of hypotheses about the imaged anatomy. The prototype system for hand radiograph analysis accurately segments normal and somewhat degenerated hand anatomy. Results are shown of the ability of the automated system to 'fail soft', recognizing when segmentation is inadequate for accurate measurement. This self evaluation capability improves reliability of measurements for potential clinical use. PMID- 8220689 TI - The identification of subcellular fractions from mammalian cells. PMID- 8220690 TI - Biomembrane protocols. I. Isolation and analysis. PMID- 8220691 TI - The isolation of membranes from bacteria. PMID- 8220692 TI - Isolation and purification of functionally intact chloroplasts from leaf tissue and leaf tissue protoplasts. PMID- 8220693 TI - Isolation and purification of functionally intact mitochondria from plant cells. PMID- 8220694 TI - Isolation of cholinergic-specific synaptosomes by immunoadsorption. PMID- 8220695 TI - Analysis of phospholipids by high performance liquid chromatography. PMID- 8220696 TI - Measurement of cholesterol in membranes. PMID- 8220697 TI - Analysis of fatty acids by gas liquid chromatography. PMID- 8220698 TI - The isolation of nuclei and nuclear membranes from rat liver. PMID- 8220699 TI - Separation and analysis of membrane proteins by SDS-polyacrylamide gel electrophoresis. PMID- 8220700 TI - Isolation of a membrane-bound enzyme, 5'-nucleotidase. PMID- 8220701 TI - The isolation of membrane proteoglycans. PMID- 8220702 TI - Analysis of sugar sequences in glycoproteins by glycosidase digestion and gel filtration. PMID- 8220703 TI - The extraction and analysis of glycosphingolipids. PMID- 8220704 TI - Isolation of mitochondria, mitochondrial membranes, lysosomes, peroxisomes, and Golgi membranes from rat liver. PMID- 8220705 TI - Continuous-flow electrophoresis. Application to the isolation of lysosomes and endosomes. PMID- 8220706 TI - Fractionation of a microsomal fraction from rat liver or cultured cells. PMID- 8220707 TI - Isolation of plasma membrane sheets and plasma membrane domains from rat liver. PMID- 8220708 TI - The isolation of clathrin-coated vesicles and purification of their protein components. PMID- 8220710 TI - Protocols in molecular parasitology. PMID- 8220709 TI - Isolation of membranes from tissue culture cells. PMID- 8220711 TI - The extraction and purification of DNA and RNA from in vitro cultures of the malaria parasite Plasmodium falciparum. PMID- 8220712 TI - Culturing and biological cloning of Trypanosoma cruzi. PMID- 8220713 TI - Application of DNA and whole organisms to filter supports for DNA probe analysis: dot, slot, and touch blotting. PMID- 8220714 TI - The development and use of repetitive sequences as DNA probes for parasite detection and species identification. PMID- 8220715 TI - Analysis of the DNA of parasitic protozoa by flow cytometry. PMID- 8220716 TI - PCR for low-level detection of malaria parasites in blood. PMID- 8220717 TI - PCR methods for identification of point mutations and gene rearrangements. PMID- 8220718 TI - Antigenic typing of field isolates of Plasmodium by DNA techniques. PMID- 8220719 TI - Ribosomal RNA probes for detection and identification of species. PMID- 8220720 TI - Rapid sequencing of parasite rRNA. PMID- 8220721 TI - Antibody screening of expression libraries. PMID- 8220722 TI - In vitro cultivation and biological cloning of Leishmania. PMID- 8220723 TI - Antibody select procedure for characterization of expression clones. PMID- 8220724 TI - Isolation of parasite genes using synthetic oligonucleotides. PMID- 8220725 TI - Transfection of Leishmania and Trypanosoma brucei by electroporation. PMID- 8220726 TI - Two-dimensional polyacrylamide gel electrophoresis. PMID- 8220727 TI - Isoenzyme electrophoresis for parasite characterization. PMID- 8220728 TI - The surface labeling of schistosomes. PMID- 8220730 TI - Immunoblotting and enzyme-linked immunosorbent assay. PMID- 8220729 TI - Monoclonal antibody affinity chromatography. PMID- 8220731 TI - Immunofluorescence of parasites. PMID- 8220732 TI - Immunoelectron microscopic localization of antigens in malaria parasites. PMID- 8220733 TI - Simple in vitro cultivation of the malaria parasite Plasmodium falciparum (erythrocytic stages) suitable for large-scale preparations. PMID- 8220734 TI - Preparation of blotted membrane for protein microsequencing. PMID- 8220735 TI - Synchronization and cloning of malaria parasites. PMID- 8220736 TI - The culture and preparation of gametocytes of Plasmodium falciparum for immunochemical, molecular, and mosquito infectivity studies. PMID- 8220737 TI - The culture of Schistosoma mansoni and production of life cycle stages. PMID- 8220738 TI - DNA sequencing protocols. PMID- 8220739 TI - DNA sequencing. PMID- 8220740 TI - Dideoxy sequencing reactions using sequenase version 2.0. PMID- 8220741 TI - Dideoxy sequencing reactions using Taq polymerase. PMID- 8220742 TI - Pouring linear and buffer-gradient sequencing gels. PMID- 8220743 TI - Electrophoresis of sequence reaction samples. PMID- 8220744 TI - Plasmid sequencing. PMID- 8220745 TI - Plasmid sequencing. PMID- 8220746 TI - Direct sequencing of inserts cloned into lambda vectors. PMID- 8220747 TI - Cosmid sequencing. PMID- 8220748 TI - Genomic sequencing. PMID- 8220749 TI - Sequencing of double-stranded PCR products. PMID- 8220750 TI - Sequencing double-stranded linear DNA with sequenase and [alpha-35S] dATP. PMID- 8220751 TI - Solid phase PCR sequencing of biotinylated products. PMID- 8220752 TI - Cycle sequencing. PMID- 8220753 TI - Direct blotting electrophoresis. PMID- 8220754 TI - Multiplex DNA sequencing. PMID- 8220755 TI - Cloning into M13. PMID- 8220756 TI - DNA sequencing by chemiluminescent detection. PMID- 8220757 TI - Reverse sequencing of M13 cloned DNA. PMID- 8220758 TI - Terminal labeling of DNA for Maxam and Gilbert sequencing. PMID- 8220759 TI - DNA sequencing by the chemical method. PMID- 8220760 TI - DNA sequencing by chemical degradation using one, two, and four different fluorophores. PMID- 8220761 TI - Linear amplification sequencing with dye terminators. PMID- 8220763 TI - Transfection of E. coli with M13 DNA. PMID- 8220762 TI - Sequencing reactions for the applied biosystems 373A Automated DNA Sequencer. PMID- 8220764 TI - Sequencing reactions for ALF (EMBL) automated DNA sequencer. European Molecular Biology Laboratory. PMID- 8220766 TI - M13 phage growth and single-strand DNA preparation. PMID- 8220765 TI - Sequencing using the Du Pont Genesis 2000 DNA Analysis System. PMID- 8220767 TI - The use of robotic workstations in DNA sequencing. PMID- 8220768 TI - M13 phage growth and DNA purification using 96 well microtiter trays. PMID- 8220769 TI - Generation of random fragments by sonication. PMID- 8220770 TI - Generation of a nested set of deletions using exonuclease III. PMID- 8220771 TI - Sequential deletions of single-stranded DNA. "Cyclone sequencing". PMID- 8220772 TI - Subcloning for DNA sequencing. PMID- 8220773 TI - Sequencing using custom designed oligonucleotides. PMID- 8220774 TI - Dideoxy sequencing reactions using Klenow fragment DNA polymerase 1. PMID- 8220775 TI - M13 cloning vehicles. Their contribution to DNA sequencing. PMID- 8220776 TI - Dideoxy sequencing reactions using T7 polymerase. PMID- 8220777 TI - Lectins: reliable differentiation markers in human oligodendrogliomas. AB - Ninety tumours classified as well-differentiated oligodendrogliomas (43 cases) and anaplastic oligodendrogliomas (47 cases) were studied with glial fibrillary acidic protein (GFAP) and bound with different lectins including: peanut agglutinin (PNA), concanavalin A (Con A), wheat germ agglutinin (WGA) and Ricinus communis agglutinin (RCA-1). PNA has shown high affinity to cellular membranes of well-differentiated oligodendrogliomas. The affinity decreases with the cellular differentiation to astrocytic lines and is lost in anaplastic oligodendrogliomas. Con A labelling is restricted to anaplastic oligodendroglioma cells and to reactive astrocytes showing a predominant somatic (cytoplasmatic) pattern of staining. Our findings showed that a combined search with GFAP and lectins may allow an accurate grading of oligodendrogliomas. PMID- 8220778 TI - Lectin histochemistry of dermoid and epidermoid cysts of the central nervous system. AB - The binding affinities of seven lectins to four epidermoid and three dermoid cysts were studied and the results were compared with the epidermis and the oral mucosa. The reactivity of four epidermoid cysts of the fourth ventricle and the frontal lobe, and one dermoid cyst in the sylvian fissure was consistent with that of the skin, whereas the binding profile of one parasellar dermoid cyst was identical to that of the keratinized oral mucosa. Another parasellar dermoid cyst also showed close similarity to the buccal mucosa. These results indicate that cells of these cysts develop same glycoconjugates as do normal epithelial cells, and some of these cysts around the sellar region might be derived from the cells of the oral mucosa. PMID- 8220779 TI - Phenotypic alteration of glioma cells during culture. AB - Tissue specimens and culture cells from three human gliomas (two astrocytomas and one glioblastoma) were immunohistochemically investigated, using GFAP, S-100P, vimentin, FN and TNF antibodies. Primary culture consisted of two cell types, flat cells and fibrous cells. Phenotypic alternation was observed during successive subculture. Differences between fibrous cells in astrocytoma and those in glioblastoma were remarkable, while flat cells in astrocytoma and glioblastoma examined in this study, were similar. PMID- 8220780 TI - Radiosensitive squamous cell craniopharyngioma: clinical and pathological comparison with the adamantinomatous type. AB - Five patients with the squamous cell type of craniopharyngioma were examined clinically and pathologically, and comparisons were made with the adamantinomatous type of craniopharyngioma. In three patients, the tumor remaining after partial or subtotal resection was treated with conventional radiation therapy, and all three lesions disappeared completely. The squamous cell tumors consisted of basal cubodial cells and polygonal superficial cells with areas of cellular detachment and degeneration. Our findings suggest that these tumors are more radiosensitive than adamantinomatous craniopharyngiomas and represent a distinct clinical entity. PMID- 8220781 TI - Clinical, biological and histological considerations on primary intracranial germinomas in the prevalent sites. AB - Twenty-four germinomas from which tumor sites were 11 pineal, 7 suprasellar, and 6 basal ganglial and thalamic (BG-T) were subjected. Humoral tumor markers of HCG, CEA and AFP were measured and compared with immunohistochemistries for these tumor markers along with PLAP, HPL and PKK1. Humoral tumor markers and immunohistochemistries did not always comprehend to each other. Difference of the survival rates among tumor sites was thought to be attributed to the variabilities in their histological, immunochemical and flow cytometric results. PMID- 8220782 TI - Anaplastic astrocytoma in the cerebellopontine angle. AB - In this paper we present a case of glioma which was located in the cerebellopontine angle. The patient, a 3-year-old male, experienced difficulty with gait for one month before admission. He was admitted to Toyota Memorial Hospital on February 2, 1991, suffering from severe headache and vomiting. Neurological examination upon admission revealed horizontal nystagmus and ataxia. MRI revealed a mass in the cerebellopontine angle. Craniotomy was performed on February 4, 1991, and a tumor was revealed in the cerebellopontine angle. The tumor was clearly demarcated and encapsulated; the cerebellum and brainstem were compressed without damage. Most of the tumor was removed. A histopathological summary of the tumor follows. The tumor appeared as exophytic lesions on the pons, extending into the cerebellopontine angle. Tumor cells contained small round nuclei and acidophilic cytoplasm. The oncocyte, which was growing endomorphically, revealed a short-cell projection, suggesting a tendency to penetrate blood vessels. Intercellular microcystic degeneration was observed clearly, with some parts of the oncocyte forming a myxoid matrix. Immunohistochemically, most of the tumor cells reacted positively to Vimentin, but negatively to S-100 protein and GFAP. Given the pathological information, the tumor was interpreted as anaplastic astrocytoma. Postoperative radiation therapy was performed, but the patient died four months later because the tumor had spread to the brainstem. In this paper we discuss the differential diagnosis of the cerebellopontine angle tumor and the appearance of anaplastic astrocytoma as exophytic lesions on the pons and the spread of the tumor into the cerebellopontine angle. PMID- 8220783 TI - Immunohistochemical studies on choroidal epithelial cyst and neuroepithelial cyst: in comparison with normal choroid plexus and ependyma. AB - Immunohistochemical evaluations with antibodies against S-100 protein, glial fibrillary acidic protein, cytokeratin and prealbumin were applied to 2 intracranial cysts of choroidal epithelial and neuroepithelial origin, and to the normal choroid plexus and ependyma. All specimens showed various immunoreactivities with the 4 antigens: especially, prealbumin-positive cells were detected in normal choroid plexus, part of normal ependyma, and part of choroidal epithelial cyst. The epithelium of neuroepithelial cysts presumably has multipotentials in cellular differentiation. PMID- 8220784 TI - Clinicopathological study on craniopharyngioma with sebaceous differentiation. AB - We presented 2 cases of craniopharyngioma, which showed sebaceous differentiation. Both cases shared the following features: 1) neuroradiological detection of a cystic tumor accompanied by calcification, 2) clear yellowish fluid detected in the cyst during surgery, 3) histological rating as an adamantinomatous type, 4) the presence of a nest-like distribution of multivacuolated cells in the innermost layer of the cyst. PMID- 8220785 TI - Autoradiographic analysis of (14C-methyl)-L-methionine uptake in transplanted rat brain tumors. AB - To elucidate metabolic pathways of methionine in brain tumors, we studied autoradiographic images of (14C-methyl)-L-methionine (14C-Met) in transplanted rat brain tumors. 14C-Met accumulated in the tumor almost twice as much as in the gray matter 90 min after injection. The acid-insoluble fraction, which was bound to macromolecules, comprised 84% the total count of the accumulated tracers in the tumor. Analysis of 14C-Met tracer accumulation is informative for understanding amino acid metabolism in brain tumors. PMID- 8220786 TI - Medulloblastoma associated with malignant rhabdoid tumor of the kidney: case report. AB - The authors report a case of 6-month-old boy with cerebellar medulloblastoma associated with malignant rhabdoid tumor of the kidney, presenting with an abdominal mass, large head and projectile vomiting. Following removal of the renal tumor, the mass arising from the superior vermis about 6 cm in diameter was removed by a combined right occipital transtentorial and suboccipital approach. The patient had been well for 3 months after surgery followed by chemotherapy but died eventually of tumor recurrence in the abdomen. Reported cases of malignant rhabdoid tumor of the kidney associated with brain tumor are reviewed and its characteristics are discussed. PMID- 8220787 TI - Ossifying cementicular fibroma of the orbitofrontal bone in a child: case report. AB - A rare case of ossifying cementicular fibroma of the left orbitofrontal bone that developed in a 12-year-old boy is presented. A hard, painless mass that was incidentally noticed gradually enlarged over 2 years. Skull X-rays showed a well demarcated lesion with mixed sclerotic and osteolytic radiolucent changes in the left orbitofrontal bone. Computed tomography revealed an expansile intradiploic multilocular mass that was separated by bony trabeculae. T1-weighted magnetic resonance imaging demonstrated a multi-cystic iso-intense mass with homogeneous contrast enhancement. Left external carotid angiograms revealed a vague tumor stain that was mainly fed by the middle meningeal artery. Systemic bone scintigrams revealed a single abnormal uptake in the lesion. The skull tumor was totally removed. Histological examination demonstrated two different characteristic findings that were composed of fibrous dysplasia and cementifying fibroma, although most of the tumor appeared to be a highly cementicular form of fibro-osseous lesion. The pathological diagnosis was a cementicular variant of fibrous dysplasia. PMID- 8220788 TI - A case of sellar chordoma mimicking a non-functioning pituitary adenoma with survival of more than 10 years. AB - A rare case of sellar chordoma occurring in a 67-year-old woman who survived for more than 10 years is presented. Clinical signs and symptoms masqueraded as a non functioning pituitary adenoma with visual disturbance and hypopituitarism. Initial computed tomography (CT) showed an intrasellar mixed dense mass with suprasellar extension, accompanied by non-homogeneous contrast enhancement. Partial removal of the mass was accomplished via right fronto-temporal craniotomy. Histological examination revealed a typical chordoma with no malignancy. After postoperative irradiation, the patient was discharged with clinical improvement. Serial CT and magnetic resonance imaging 8 years after treatment disclosed a regrowth of the intrasellar lesion, which extended to the sphenoid sinus and clivus, accompanied by non-homogeneous contrast enhancement. The patient underwent subtotal removal of the recurrent tumor through a sublabial transsphenoidal approach. Histological examination confirmed the previous diagnosis. Immunohistochemical study demonstrated positive cytoplasmic expression of vimentin, epithelial membrane antigen, keratin and S-100 protein, in contrast with a lack of appearance of carcinoembryonic antigen. After reoperation, she completely recovered and has survived for more than 10 years with good quality of life. PMID- 8220789 TI - Immunohistochemical study of copper and zinc-superoxide dismutase (Cu, Zn-SOD) in meningiomas. AB - A localization of copper and zinc-superoxide dismutase (Cu, Zn-SOD) in meningiomas was examined with immunohistochemical technique. Meningiomas include five meningotheliomatous, five transitional, two fibroblastic, and one of each angioblastic, malignant and hemangiopericytic types. The Cu, Zn-SOD localized diffusely in the cytoplasm of tumor cells and in the endothelial cell of vessels and neutrophils, macrophages and red blood cells in the vessels or in the tumor tissue. The Cu, Zn-SOD content in meningiomas was markedly higher in angioblastic, malignant and hemangiopericytic than in meningotheliomatous, transitional and fibroblastic types. PMID- 8220790 TI - Poorly differentiated orbital rhabdomyosarcoma with intracranial extension: a case report. AB - The authors report a case of poorly differentiated orbital rhabdomyosarcoma with intracranial extension in a 5-month-old female. The clinical and pathologic characteristics are discussed with reference to the differential diagnosis. PMID- 8220791 TI - Origin of craniopharyngioma: an electron microscopic study. AB - To investigate the possibility that the two subtypes of craniopharyngioma, adamantinous type and squamous papillary type, are tumors arising from different origins, the authors conducted an electron microscopic study in 10 cases of craniopharyngioma. Light microscopic study revealed distinct differences between the two subtypes, but their ultrastructure was fundamentally similar. In this report, the authors propose an explanation of the origin of these subtypes to account for the findings. PMID- 8220792 TI - Malignant nerve sheath tumor of the jugular foramen followed by meningeal dissemination in a patient with neurofibromatosis. AB - A 20-year-old female with von Recklinghausen's disease became aware of hoarseness and dysphagia with a right neck mass, and received subtotal resection of a jugular foramen neurofibroma. One year later the jugular foramen tumor recurred and pathologic examination at excision showed a malignant transformation. Despite postoperative chemotherapy and radiotherapy, it recurred soon and followed the metastatic cerebral tumor by meningeal dissemination. Malignant nerve sheath tumors of the jugular foramen are very rare and the clinical course and pathological pictures of this tumor are reviewed. PMID- 8220793 TI - Malignant astrocytoma with extracranial metastases: a case report. AB - A 41-year-old man was referred to this hospital after being diagnosed as suffering from malignant astrocytoma. In spite of two operations and subsequent chemoradiation therapy, the patient died 20 months after the onset of disease. At autopsy, the authors found skin invasion of tumor at left temporal region, softening of the brain and subarachnoid hemorrhage at the base of the brain. Yellowish-white bulging lesions were found at the bottom of left lung, diaphragm and parietal pleura. Histologically, primary lesion showed features of anaplastic malignant astrocytoma. Subarachnoid dissemination was noted at the base of the brain and in the spinal canal. Invasion into the vessels were observed both at the primary site and at the base of the brain. Glial fibrillary acidic protein positive spindle-shaped tumor cells proliferation was seen in the metastatic lesions. This case was diagnosed as malignant astrocytoma with remote extracranial metastases. PMID- 8220794 TI - Histological and ultrastructural findings of benign intracranial cysts. AB - Since benign intracranial cysts have similar clinical and neuroradiological imaging appearances, it is difficult to distinguish between these cysts. Thus an electron microscopic study plays an important role in definite diagnosis of these non-tumorous benign cystic lesions, and also in determination of their origins. Arachnoid cysts occurring in 5 patients, epithelial cysts occurring in 2 patients, 2 cases of Rathke's cleft cysts, and one case of pineal cyst, were electron- and light-microscopically observed, and their characteristic features and differences were described. The structure of the arachnoid cyst wall was similar to that of normal arachnoid membrane. The inner surface of the arachnoid membrane. The inner surface of the arachnoid cyst wall was formed of one or several layers of arachnoid cells with slender processes, which contained large extracellular spaces, but not microvilli. As for epithelial cysts, microvilli and surface-coating material were present on the surface of epithelial cysts, and a well-developed basement membrane was continuously observed under the epithelial cells. Rathke's cleft cysts, which may basically be classified as epithelial cysts, are lined by columnar or cuboidal ciliated epithelium. The wall of the pineal cyst is composed of normal pineal tissue, and there are three layers, which have a lining of glial cells in the inner layer, a few sheets of pineal cells in the middle layer, and a fibrous capsule in the outer layer. PMID- 8220795 TI - Secretory meningioma: a case report with immunohistochemical and ultrastructural study. AB - Secretory meningioma is a new concept proposed with the progress on immunohistochemistry, and has not sufficiently been discussed in the literature. We report a case with this rare type of meningioma, in a 54-year-old female. The tumor had hyaline inclusions that showed not only carcinoembryonic antigen (CEA) on immunohistochemical study but some secreting organelles on ultrastructural study. The value of these findings are emphasized in the histological diagnosis for secretory meningioma. PMID- 8220796 TI - Vacuolated meningioma with secretory features: a case report. AB - We report a case of meningioma with secretory features in a 47-year-old woman. The left frontal tumor mass had already eroded the overlying bone at the time of presentation. When respected, the mass showed a gelatinous appearance. Histologically, marked vacuolar change was seen in the stroma. Whorls, psammoma bodies, and pavement arrangement of nuclei were also noted. Periodic acid-Schiff stain revealed cytoplasmic granules, which were digested with diastase. Alcian blue diffusely stained the cytoplasm of the tumor cells. Electron microscopy showed numerous small vacuoles and cored vesicles in the cytoplasm. Glycogen granules were poorly observed. These features indicate the mild secretory nature of this vacuolated meningioma. PMID- 8220797 TI - Hydrolysed cow's milk formulae. Allergenicity and use in treatment and prevention. An ESPACI position paper. European Society of Pediatric Allergy and Clinical Immunology. PMID- 8220798 TI - Food and behaviour. Allergy, intolerance or aversion. PMID- 8220799 TI - Prevalence of atopic disease among Danish school children. AB - As a result of a 1990 survey by questionnaire, the symptoms of atopy among all 4,952 school children aged 5 to 16 years in the municipal district of Viborg, Denmark, were registered. Random checks, made among children who were recorded as having symptoms, and others who were recorded as having none, accorded well with the information supplied by the parents about symptoms and the clinical diagnosis of a specialist; 10.5% of all school children had rhinitis, 7% had atopic eczema, 3.2% had urticaria and 4.5% had asthma; 1/4 of all those questioned had shown symptoms within the last year, and a further 13% of all the children were reported as having had atopic symptoms that had disappeared more than a year previously. Of the children showing symptoms within the last year before the survey, 2/3 had gone to a doctor. Of the children with present symptoms, largely asthma, 1/4 had been referred to a hospital allergy clinic. For 1/3 of the children with present symptoms, these had led to no contact with a doctor. Of the cases with present symptoms, 6.5% had had contact with natural healers or chiropractors. Rhinitis and asthma were most frequent among boys, while atopic eczema was most frequent among girls. For both sexes, the frequency of rhinitis increased during their years at school, while the frequency of skin symptoms fell. PMID- 8220800 TI - Prevalence of intolerance to food additives among Danish school children. AB - The prevalence of intolerance to food additives was assessed in a group of unselected school children aged 5-16 years. A study group of 271 children was selected on the basis of the results of a questionnaire on atopic disease answered by 4,274 (86%) school children in the municipality of Viborg, Denmark. The children in the study group followed an elimination diet for two weeks before they were challenged with a mixture of food preservatives, colourings and flavours. The challenge was open and the additives were prepared as a fizzy lemonade. If the open challenge was positive, a double-blind placebo controlled challenge with gelatine capsules was performed. The study included 281 children, 10 were excluded, and the remaining 271 children were given the open challenge (98 healthy controls and 173 with atopic symptoms). The open challenge was negative in all 98 healthy control children who had not reported any atopic symptoms. Of the 173 children reporting present or previous atopic disease 17 had a positive open challenge. Of these 17 children 1 experienced gastrointestinal symptoms, 13 reacted with aggravation of atopic eczema, and 3 with urticaria. Twelve of these 17 children went through the double-blind challenge which was positive in 6 cases. Five of these 6 children had positive reactions to synthetic colourings and 1 to citric acid. No serious reactions were seen. Based upon calculations of the results from this study and an earlier multi-center study in children referred to hospital clinics, the prevalence of intolerance to food additives in school children is estimated to be 1-2%. PMID- 8220801 TI - Atopic family history. Validation of instruments in a multicenter cohort study. AB - In order to identify newborns at risk for atopic diseases, we developed a family questionnaire and selected specific answers which were suitable to identify atopic family members. The validity of the questionnaire was evaluated by the Phadiatop test results of 793 mothers and 353 fathers. As both screening instruments do not measure the same, the Phadiatop test identifies sensitization to inhalant allergens and the history reflects the clinical manifestation of atopic disease, the agreement between sensitization and manifestation is incomplete. Sensitivity and specificity of the questionnaire screening conditions to reproduce the Phadiotop test result was 64% and 84% for mothers, and 58% and 88% for fathers, respectively. The relative risk for lifetime prevalence of atopic manifestations in Phadiatop positive over negative mothers was calculated to be 3.88 (95% confidence interval = 3.12 to 4.81), and for Phadiatop positive over negative fathers to amount 4.84 (95% confidence interval 3.25 to 7.23). A few relevant answers of 20 were identified by logistic regression analysis to predict the Phadiatop test result nearly, as well as the total questionnaire. PMID- 8220802 TI - Vacuum cleaning decreases the levels of mite allergens in house dust. AB - To investigate the capacity of chemical treatment of surfaces and the difference in capacity among common vacuum cleaners to reduce mite allergen content in house dust, we recruited 52 families with allergic children. Ten families used their central vacuum cleaners. Forty-two families were randomly divided into four groups with 10 or 11 families in each. These families used either new vacuum cleaners with either HEPA (High Efficiency Particulate Air) or micro-filters, or their own vacuum cleaners with either tannic acid or placebo. Dust samples were collected from carpets and upholstered furniture in the living rooms and from the mattresses of the children at Days 0, 7, 21, and 35. Der pI and Der fI allergens were determined by sandwich ELISA. After one week, tannic acid reduced the concentration of mite allergens/g of dust and the total amount/sampling area by 30% and 34%, respectively (p < 0.05), but there was no significant decrease in relation to placebo. After 5 weeks, central, HEPA- and micro-filter vacuum cleaners decreased the mite allergen concentration by 10-50% (p < 0.05) and the total amount of mite allergen from the investigated areas by 50-85% (p < 0.01). In relation to the placebo group the decrease was significant for HEPA- and micro filter vacuums (p < 0.05). The total amount of mite allergens/sampling area was more significantly (p < 0.05) reduced than the concentration/g of dust. We conclude, that tannic acid reduces mite allergen concentrations in dust and total amount/sampling area for a short period of time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8220803 TI - Nasal powder administration of budesonide for seasonal rhinitis in children and adolescents. AB - The effect of intranasal budesonide delivered from a new dry powder inhaler (Rhinocort Turbuhaler) was evaluated in a randomized, double blind, group parallel study of 83 children aged 4-16 years with seasonal allergic rhinitis. The doses 100 and 200 micrograms taken once daily in each nostril were compared with placebo. A one week run-in period was followed by a 4 weeks treatment period. Diary recordings of nasal symptoms, p.r.n. use of terfenadine tablets were made at home and rhinoscopy performed at hospital before and at the end of each treatment period. Total or substantial control of symptoms was achieved in 41% of the children in the 400 micrograms group (p < 0.05 compared with placebo), in 33% in the 200 micrograms group (p = 0.07 compared with placebo) and in 14% in the placebo group. The 400 micrograms but not the 200 micrograms group was statistically significantly better than placebo for most of the clinical effect parameters studied. The treatment was well tolerated and a battery of laboratory tests including cortisol excretion in the urine was not influenced by the various treatments. These results indicate that 400 micrograms budesonide given once daily from a Rhinocort Turbuhaler is clinically effective and safe in children with rhinitis. Further studies are needed to evaluate if this inhaler is preferable to other inhalers currently used for the treatment of rhinitis. PMID- 8220804 TI - Cetirizine for seasonal allergic rhinitis in children aged 2-6 years. A double blind comparison with placebo. AB - A total of 107 children of both sexes between 2 and 6 years of age with pollen induced seasonal allergic rhinitis (SAR) were entered in a multicentre study of double-blind parallel group design in which the effects of 5 mg cetirizine, given as drops from a solution containing 10 mg/ml once daily each evening for two weeks, were compared with those of identical placebo. Sneezing, rhinorrhea, nasal obstruction and nasal and ocular pruritus were the symptoms evaluated by means of symptom scores by investigators and, on daily record cards, by parents. Investigators also made a global evaluation at the end of treatment. Cetirizine was more active than placebo for each symptom evaluated both by parents and investigators. There were significant by more (p = 0.002) days during which symptoms were absent or mild, in the cetirizine than in the placebo group. When the maximum symptom scores rated by investigators at each visit were compared, the difference in favour of cetirizine at the end of treatment was statistically significant (p = 0.04). Global evaluation by investigators of changes in symptoms at the end of the study showed an improvement in both groups which was significantly greater with cetirizine, providing excellent or good improvement in 34/54 patients compared with 25/53 patients on placebo (p = 0.039). Tolerance was good. Three patients on cetirizine and none on placebo experienced mild somnolence. PMID- 8220805 TI - Use of hydrolyzed cow's milk formulae for prevention of early sensitization and signs of atopy must be further documented. PMID- 8220806 TI - Judges as gatekeepers: guidelines in court. Interview by Paul M. Schyve. PMID- 8220807 TI - Decision making with pregnant patients: a policy born of experience. AB - One hospital has developed an institutional policy to guide decision making between physicians and pregnant patients. Read the policy and a description of how it was developed by the ethics committee at the George Washington University Hospital. PMID- 8220809 TI - The legal implications of clinical practice guidelines: a workshop sponsored by the Association of Academic Health Centers. PMID- 8220808 TI - Letter from the Interest Group on Health Services Research, Association of Academic Health Centers. AB - Although guidelines have existed in various forms for many years, increased interest in guidelines has sparked myriad concerns among health care professionals, policymakers, and the broader public. Among these concerns are a number of legal questions that to date have not received sufficient notice or study. PMID- 8220810 TI - Clinical practice guidelines: policy issues and legal implications. AB - The growing interest in clinical practice guidelines is raising a number of complex legal questions that have yet to be widely examined. Could physicians be held liable if they disregard good medical judgement in favor of following cost containment guidelines? To what extent should developers and implementers of guidelines be granted civil immunity from liability? How will judges respond to guidelines as evidence, and how will they react when various guidelines conflict? PMID- 8220811 TI - Overview: prospects and options for local and national guidelines in the courts. AB - Local adaptation of national guidelines appears to be widespread, but it is little examined in terms of its extent, appropriateness, and impact. But the worst situation for patient and practitioner would be when a good national guideline exists but a bad local guideline guides practice. PMID- 8220812 TI - Overview of legal issues. AB - Evidence suggests that compliance with guidelines will affect malpractice litigation. One point often overlooked is that guidelines can provide both exculpatory and inculpatory evidence. PMID- 8220813 TI - Use of practice parameters as standards of care and in health care reform: a view from the American Medical Association. AB - Guidelines that are standards of care could limit the creative initiatives to find ways of caring for patients which do not conform to traditional medical practice but result in good outcomes. Physicians would likely feel hemmed in by guidelines that constitute an express legal standard of care and would be afraid to deviate from them except under the auspices of carefully controlled research protocols. PMID- 8220814 TI - The use of guidelines to reduce costs and improve quality: a perspective from the insurers. AB - The natural concern is the abuse of guidelines--most commonly, from using them as rules, either applying them incorrectly or attempting to micromanage care through them. Both corporate customers and insurers are greatly concerned about these issues and have policies to avoid such abuse. PMID- 8220815 TI - A view from the Agency for Health Care Policy and Research: the use of language in clinical practice guidelines. AB - "Good medicine is good law" is the first principle for writing good guidelines. The most important legal inquiry will be whether the language specifies mandatory or persuasive actions. Ordinary words may be hazardous when they are used ambiguously and create more than one meaning. That may mean a jury will decide what was meant. PMID- 8220817 TI - A case study in progress: practice guidelines and the affirmative defense in Maine. AB - To encourage physicians to practice less defensively, the Maine Medical Liability Demonstration project adopted the controversial "affirmative defense" to combat runaway malpractice liability by granting immunity to physicians who follow practice guidelines. Despite challenges from trial lawyers, physicians are beginning to accept the program. PMID- 8220816 TI - Assisting judges in screening medical practice guidelines for health care litigation. AB - In a far-reaching decision about scientific evidence in federal cases handed down at the very end of the 1992-1993 term, the U.S. Supreme Court implicated medical practice guidelines in indirect, profound, yet highly uncertain ways. In Daubert v Merrell Dow Pharmaceuticals, Inc the Court ruled that judges must actively screen novel scientific evidence to assure both its relevance and its reliability. PMID- 8220818 TI - Ultrastructural study of glomerular basement membrane in diabetic rats by quick freezing and deep-etching method. AB - The three-dimensional ultrastructure of glomerular basement membrane (GBM) in streptozotocin (STZ)-induced diabetic rats was examined by quick-freezing and deep-etching method. In three layers of the GBM of control rats, the outer and inner layers were formed by files of perpendicular fibrils, which connected the epithelial or endothelial cell surfaces with meshwork structures of the middle layer. In the diabetic rats, the inner layer was diffusely enlarged and the meshwork structure of the middle layer became markedly irregular due to the rupture of fine fibrils and thickening of material adherent to the fibrils. These ultrastructural changes correspond to those of subendothelial oedema, lamellation of lamina densa and fluffy material in the GBM, as revealed on conventional ultra thin sections. It is suggested that the initial morphological change of STZ induced diabetic nephropathy is disruption of matrix fibrils in the GBM, seemingly indicating a disturbance of size and/or charge barriers. PMID- 8220819 TI - The effect of fibronectin on cytoskeleton structure and transepithelial resistance of alveolar type II cells in primary culture. AB - The cytoskeleton of alveolar type II cells on different matrices has been examined, and the bioelectric properties of these cells grown as monolayers in primary culture has be measured using Ussing-type chambers, to determine whether the extracellular matrix affects the cytoskeletal organization of alveolar type II cells and whether any such interactions influence their physiological functions. Alveolar type II cells cultured on a fibronectin substratum spread slowly over a 6-day period to produce cells of extremely large diameter. The cytoskeletal structure of these cells was characterized by a more marked accumulation of large bundles of actin and a finer network of keratin than cells grown on a collagen substratum. The transepithelial resistances of monolayers grown on a fibronectin substratum were much higher than those on a collagen substratum. These results indicate that alveolar type II cells cultured on fibronectin can form tighter, better organized and more polarized monolayers in primary culture, which suggests that fibronectin may have a physiologically important role in the maintenance of the alveolar wall. PMID- 8220820 TI - Histochemical and biochemical determination of calcium in pleomorphic adenoma. AB - Although calcification seldom occurs in pleomorphic adenoma, it often occurs in salivary glands, and so we decided to investigate the possible role of calcium in this difference. A histochemical method using glyoxal bis(2-hydroxyanil) demonstrated a small amount of calcium outlining lumina and separated cells of epithelial structures and associated with cells of myxoid and chondroid regions in pleomorphic adenoma, and a conspicuous amount in the acini of the associated salivary glands. A biochemical method using dry ashing demonstrated a significantly higher level of calcium in the glands than in pleomorphic adenoma. The results indicate that the calcium is mainly associated with secretory granules, which are scarce in pleomorphic adenoma, and with proteoglycan present intercellularly and in stromal regions of pleomorphic adenoma. The calcium in secretory granules is of possible importance in calcification in lumina and epithelium, and that bound to proteoglycan is possibly released following necrosis to be of importance in stromal calcification. However, the overall low level of calcium in pleomorphic adenoma is the likely explanation for the usual lack of calcification. PMID- 8220821 TI - In-situ polymerase chain reaction. An overview of methods, applications and limitations of a new molecular technique. AB - The in-situ polymerase chain reaction (in-situ PCR) is a novel molecular technique that combines the extreme sensitivity of the PCR with the anatomical localization provided by in-situ hybridization. A number of groups have recently reported studies using in-situ PCR for the detection of specifically amplified single-copy nucleic acid sequences in single cell preparations or low copy DNA sequences in tissue sections. In this overview, we describe the principles of in situ PCR, review the applications of this technique and discuss future aspects of in-situ PCR. We critically compare the different in-situ PCR protocols described in the literature. Emphasis is placed on the absolute requirement for controls to allow accurate interpretation of results and the possible problems and pitfalls of the in-situ PCR methods, including artefacts related to diffusion of PCR products and non-specific incorporation of labelled nucleotides into fragmented DNA undergoing repair. It is concluded that this technique will eventually play an important role in specialized diagnostic laboratories in the evaluation of viral diseases, haematological and other malignancies which have unique genetic markers. PMID- 8220822 TI - Ultrastructure of human dermal mast cells in 29 different lysosomal storage diseases. AB - The effect of lysosomal storage diseases on the ultrastructure of human mast cells has not previously been reported. Indeed, there has been little published evidence indicating that mast cells contain typical lysosomes. However, mast cell cytoplasmic granules contain hydrolases similar to those found in lysosomes, but which differ from lysosomal hydrolases in exhibiting optimal activity at higher pH. We therefore examined by transmission electron microscopy the dermal mast cells in 58 biopsies of patients exhibiting 1 of 29 different lysosomal storage diseases. We found mast cells containing abnormal lysosomes in 16 of these disorders. In 6 of these 16 diseases, the mast cells' cytoplasmic granules appeared normal. These observations indicate that human mast cells can contain lysosomes, and provide evidence that the enzymes affected by lysosomal storage diseases are active in mast cells. PMID- 8220823 TI - Role of fat-storing cells in schistosomal hepatic fibrosis of mice. AB - The involvement of fat-storing cells (FSC) in hepatic schistosomal granuloma was investigated in mice infected with Schistosoma mansoni cercariae. After infection, 24 animals were treated with s.c. injections of vitamin A in a total dose of 210,000 IU given twice a week for 3 weeks. Two other groups of 24 animals each were: a) non-infected vitamin A-treated animals and (b) untreated infected controls. Animals from all groups were killed at weekly intervals from the 5th through the 10th week following infection. Pieces of liver were examined by light microscopy and transmission electron microscopy. In all vitamin A-treated animals, FSC disclosed prominent cytoplasmic fat droplets, which permitted their prompt identification in the light and in the electron microscope. They were found in large numbers as a constituent of periovular granulomas. In infected controls, FSC were not identified in granulomas, possibly because lipid droplets disappeared during differentiation to the fibroblastic phenotype. FSC also appeared within areas of septal fibrosis. These data suggest that FSC play an important part in focal portohepatic fibrosis during granuloma formation around S. mansoni eggs in the liver of mice. PMID- 8220824 TI - Effects of hypercholesterolemia on initial and chronic phases of rat nephrotoxic serum nephritis: development of focal segmental glomerulosclerosis, analogous to atherosclerosis. AB - The effects of hypercholesterolemia on both the initial and chronic phases of rat nephrotoxic serum (NTS) nephritis have been investigated. Injury during the initial phase of NTS nephritis in hypercholesterolemic rats maintained on a cholesterol-supplemented diet (Group 2) was characterized by segmentally accentuated accumulations of vacuolated cells with lipid droplets (foam cells) in the glomeruli, while the kidneys of rats fed a standard diet (Group 1) revealed only mild intracapillary cell proliferation. Immunoelectron microscopy showed that the foam cells observed in Group 2 rats were largely derived from macrophages. The glomerular macrophage number, defined by the number of ED1 positive cells per glomerulus, was significantly higher in Group 2 than in Group 1 animals at days 5-6 (3.4 +/- 1.4 in Group 1 against 6.3 +/- 1.0 in Group 2; p < 0.01) as well as at days 21-28 (5.5 +/- 2.6 in Group 1 against 10.9 +/- 2.8 in Group 2; p < 0.01). In contrast, the numbers of OX19-positive T-lymphocytes and OX33-positive B-lymphocytes were similar in both groups. In the chronic phase of NTS nephritis at week 20, semiquantitative evaluation of the glomerular lesions disclosed more severe focal segmental glomerulosclerosis (FSGS) in Group 2 compared with Group 1 animals (glomerular injury score: 14 +/- 10 in Group 1 against 73 +/- 17 in Group 2; p < 0.01). Accumulations of lipid and foam cells were invariably seen in the sclerotic foci of Group 2 animals. The results indicate that hypercholesterolemia played an important role in the accelerated development of FSGS in rat NTS nephritis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8220825 TI - [The fatty acid composition of the cellular structures of Yersinia pestis]. AB - Fatty acid composition of lipopolysaccharides (LPS) and cytoplasmic membranes (CPM) of Yersinia pestis [correction of the plaque microbe] has been studied. Concentration of certain acids in the LPS content proved sharply different and thus the strains were separated into groups. High content of 3-oxymiristinic acid was a distinction of vaccine strains. Virulent strains of one of the groups regularly differed from the rest of virulent strains by high content of laurinic acid. Laurinic and two nonidentified acids, which were not found in the virulent cultures, were present in CMP lipids of vaccine strains. PMID- 8220826 TI - [Soluble and semisoluble lectins from mollicutes and their possible functions]. AB - When studying mollicute lectins it was established that Acholeplasma laidlawii PG 8 synthesizes two half-soluble lectins one of which is specific to N-acetyl-D glucosamine, and the other--to D-glucosamine.HCl; phytopathogenic strain A. laidlawii var. granulum 118 produced 4 lectins one of which is soluble and specific in respect to fructose-1.6-diphosphate, the rest three lectins are half soluble and specific to one of the sugars--D-galactosamine.HCl, rafinose and D glucosamine.HCl. In Mycoplasma pneumoniae FH all the four lectins found in the culture liquid have been classified as half-soluble, specific to one of carbohydrates--D-galactosamine.HCl, talose, N-acetyl-neuramine acid and D glucose; M. capricolum Cal. Kid. synthesizes four lectins; two of them being defined as soluble (one of the lectins is, respectively, specific to talose and D glucosamine.HCl, two others, as half-soluble, specific to one of sugars--rafinose or D-glucose. The results obtained permit a conclusion to be made that the half soluble lectins of mollicutes, on the one hand, are the factors of adhesion on the corresponding organs of macroorganism and, on the other hand take part in the transport of substances from without into the microorganism cell. Soluble lectins determine pathogenicity of mollicutes and form with half-soluble lectins a single chain to providing the mycoplasma cells with nutrients and to protect them from the action of the macroorganism immune system. PMID- 8220827 TI - [The resistance of the DNA of cyanophage LPP-3 to the action of different restriction endonucleases]. AB - Data on the study of structure peculiarities of cyanophage LPP-3 DNA are presented in the work. The length of cyanophage DNA calculated by means of the enzymatic hydrolysis by restrictases is 40 +/- 3.5 thou. pairs of bases. Cyanophage LPP-3 DNA was hydrolysed by more than 50 different restrictases. As a result of screening it was found out that the great number of restrictases, which recognized hexanucleotide sequences did not hydrolyze DNA of cyanophage LPP-3. A considerable deviation of the number of the observed sites of restriction from their theoretically expected number for restrictases Hae III and Cfr 131 was established. Restrictases-isoschisomeres with different sensitivity to the methylation of the recognition sites--Msp I, Hpa II and Sau 3A, MboI and DpnI were used to check the availability of methylated bases in LPP-3 DNA. Absence of methylated adenine in the site GATC and methylated cytosine in the second position of the site CCGG were established. The results obtained permit supposing that the expressed counterselection by the sites of recognition of many restriction endonucleases takes place in cyanophage LPP-3 DNA. It is supposed that apparently, this method of protection of its genome in LPP-3 is one of most important but the inconsiderable percentage of site-specific methylation of the virus DNA cannot be completely excluded. PMID- 8220828 TI - [The participation of the neuraminidase of Yersinia pestis in the transmission of the transmembrane signal]. AB - Neuraminidase isolated from Yersinia pestis [correction of the plaque microbe] and purified has been studied for its effect on the peritoneal leukocytes of the white mice with results presented. Neuraminidase exerted the insulin-like effect on the peritoneal leukocytes of the white mice and this effect depended on the used dose. The priming of leukocytes by neuraminidase preparation was accompanied by the sharp increase of hydrogen peroxide production under effect of the capsule antigen of the plaque microbe and, apparently, by the appearance of additional binding sites for the adhesion piles of the plaque agent. PMID- 8220829 TI - [The body's nonspecific resistance factors in a localized experimental infection caused by Bacteroides]. AB - Under local experimental infection induced by bacteroides of Fragilis group the factors of nonspecific organism resistance take an active part in the inflammation process. Phase changes of the state of monocytic-phagocytic and complement systems were observed. Peripheral blood leukocyte phagocytic activity decreased at the primary stage, then followed the complement activation by the alternative pathway mainly. The increase of phagocytic and metabolic activities of those phagocytes taking part in the inflammation, the complement at this stage is activated due to the classical pathway while being at the stage of clinic manifestation. Levels of antibody titres are also increasing. PMID- 8220830 TI - [The carriage of Staphylococcus aureus among different population groups]. AB - The level of carriage of Staphylococcus aureus in three biotopes (nose, fauces, hand skin) and categories of carriers among students and teachers of the Medical Institute, medical personnel of surgical department and maternity hospital as well as the workers of combine building plant have been studied. The higher percentage of Staphylococcus aureus carriers was established in the nasal cavity of the maternity hospital personnel (p < 0.001) and on the hand skin of the personnel of surgical stationary (p < 0.05) as compared to the students and teachers. Carriage frequency of Staphylococcus aureus among the workers several times exceeded its level in the control group. The strains of the third phage group predominated among the personnel of surgical department, those of the second and fourth phage, groups, among the maternity hospital workers. Representatives of the second phage group were isolated in the plant workers. High level of carriage among the workers creates preconditions for the development of purulent diseases of the hand and fingers after microtraumas at the plant. All this dictates a necessity to eliminate Staphylococcus aureus in the carriers of all three biotopes. PMID- 8220831 TI - [The pathogenicity factors in different representatives of the genus Citrobacter]. AB - No trustworthy distinctions were revealed under the comparative analysis of properties of bacteria of Citrobacter genus from 3 studied ecological groups (from patients with diarrhea, from healthy persons and patients with extraenteric pathological process). It allows the finding of potentially partogenic bacteria of Citrobacter genus in patients with diarrhea to be considered the manifestation of dysbacteriosis. PMID- 8220832 TI - [The dynamics of the fungal mycelial content in the soils of stationary posts in a 30-kilometer zone around the Chernobyl Atomic Electric Power Station]. AB - The paper deals with mycobiota of radionuclide-polluted soils of seven stationary posts in the 30-kilometer zone of the Chernobyl NPP and one in the vicinities of Kiev (Feofaniia). The lengths of light-and dark-coloured fungal mycelium were determined in the soil under study by the method of membrane filters for 1987 1989. Data obtained have been analyzed with the account of the radioactivity level of the soil samples, season and depth of the sampling horizon. The dynamics of mycelium content in soils has been traced which evidences for its seasonal changes and prevalence of dark-pigmented mycelium at the beginning which then was changed by predominance of light-coloured mycelium. It is supposed that radioactive contamination of soil is one of considerable components of the whole complex of ecological factors which determine the dynamics of fungal mycelium content in soil. PMID- 8220833 TI - [The current concepts of the mechanisms of the therapeutic-prophylactic action of probiotics from bacteria in the genus Bacillus]. AB - Data from literature as well as the authors' experimental data on the mechanism of the effect of probiotics from aerobic sporeforming bacteria have been first generalized. The authors made a conclusion that the effect of biopreparations from bacilli is the multifactor process, when bacilli get to the gastrointestinal tract they begin producing biologically active substances which make the immediate effect on pathogenic and conditionally pathogenic microorganisms as well as activate specific and nonspecific systems of the macroorganism protection. The unknown before phenomenon of bacteria translocation is analyzed in detail as one of the most important factors of activation of non-specific resistance of the macroorganisms. Aerobic sporeforming bacteria produce a number of enzymes, amino acids, antibiotic substances and other biologically active substrates which add to the complex treatment-prophylactic effect of biopreparations from these bacteria. Prospects of the use of probiotics based on aerobic sporeforming bacteria in medicine and veterinary are considered. It is emphasized, in particular, that the potential multifactor character of the treatment-prophylactic efficiency of biological preparations from Bacillus is of positive value, since no negative effects are evoked by the known mechanisms of specific effect of aerobic sporeforming bacteria. This opens wide potentialities for improving the schemes and methods of application of probiotics from bacilli, creation of new forms of drugs on their base as well as to combine them with other preparations. PMID- 8220834 TI - Day care obstetrics. PMID- 8220835 TI - Guillain-Barre syndrome: the latest on treatment. PMID- 8220836 TI - Brain-imaging techniques: use and overuse. PMID- 8220837 TI - Herpesvirus infections in childhood: 1. AB - Infections due to herpesviruses have received increasing attention over the past decade, culminating in the isolation in 1986 of human herpesvirus 6. We examine the clinical spectrum of acquired herpesvirus infections in children and review developments in our understanding of their molecular biology, pathogenesis, treatment and prevention. PMID- 8220838 TI - Urinary urgency in women. AB - Urinary urgency is a common unpleasant symptom which may result from a variety of different pathologies. Careful patient assessment and appropriate investigation allow the cause to be elucidated and treated effectively in most cases. However, further research is necessary to increase our understanding of these disorders and to improve the quality of life of the women affected by them. PMID- 8220839 TI - Antiphospholipid antibodies in pregnancy: clinical associations. AB - Antiphospholipid antibodies are a group of pathogenic antibodies associated with adverse pregnancy outcome. Pregnancies complicated by the presence of antibodies require careful surveillance and in specific situations therapy with low-dose aspirin, corticosteroids and/or heparin. PMID- 8220840 TI - Alcohol and depression. AB - Chronic alcohol misuse lowers mood and often leads to suicide or suicide attempts. yet drinking problems are often overlooked in patients presenting with depression, or the severity of mood change is unrecognized in a known alcohol misuser. Anxiety frequently accompanies problem drinking and accentuates depression. This article considers the implications for therapy. PMID- 8220841 TI - First impressions. AB - 'During a consultation there are two people at work. While the doctor is searching for a diagnosis, the patient is quietly summing up the doctor. And it is often the patient who reaches his conclusion first.' This quote from William Evans illustrates how the consultation involves a two-way analysis by both doctor and patient. PMID- 8220842 TI - Management of acute asthma in childhood. AB - In children, acute asthma is one of the most common reasons for admission to hospital. Morbidity and mortality from asthma are unnecessarily high. It is essential that all those caring for asthma patients can accurately assess and treat an acute exacerbation of asthma. PMID- 8220843 TI - Slides in tropical medicine. PMID- 8220844 TI - Disseminated intravascular coagulation in adult-onset Still's disease with neurological, respiratory and hepatic sequelae. PMID- 8220845 TI - RHAs sweat in reforms wrangle. PMID- 8220846 TI - Aspirin, warfarin and stroke. PMID- 8220847 TI - Organization of obesity services. PMID- 8220849 TI - Lords, ladies and audit men. PMID- 8220848 TI - Pressure-controlled ventilation in asthma. PMID- 8220850 TI - A peri-implant capsule flap. AB - Spherical expanders (30 ml) were implanted under the skin vascularised by the left inferior epigastric pedicle in rats. When expansion was complete, the expander was removed and the animals divided into three groups of 15. In the first group, the floor of the capsule was simply everted. In the second group, a capsule island flap was raised; in the third group, a capsule free flap was raised, transferred to the heterolateral vessels by microanastomosis; the inner side of the various capsule flaps was covered with autologous skin graft. In the three experimental groups, there was complete "take" of the skin grafts in 80% of the animals. Pedicle or free flaps of capsular tissue may be raised and transferred safely in rats. PMID- 8220851 TI - Surface ultrastructure of human dermis and wounds. AB - Scanning electron microscopic observations of the de-epidermalized surfaces of wounds and control skin reveal the architecture of dermal collagen. At wound edges there is a transition from the normal regular rete peg structure through erratically shaped finger-like projections with buds, to the relatively flat surface of granulation tissue. A corresponding change in the ultrastructure of dermal collagen is seen from normal areas where bundles of thick fibres are made up from finer fibrils, through a transitional zone where only fine fibrils are visible, to the granulation tissue where a disorganised amorphous structure is seen. It is suggested that the fibrous tissue surface at the edges of healing wounds is extensively remodelled after it is covered by epidermal cells. PMID- 8220852 TI - The morbidity of the free vascularised fibula flap. AB - Twenty patients who had undergone free vascularised fibula flap transfers were reviewed to determine the incidence of peri and postoperative complications. The mean follow-up was 3.2 years (0.2-10); 18 grafts were successful, but required 10 re-operations for complications including 5 haematoma evacuations and 3 bone grafts and plates for delayed or non-union. The two unsuccessful fibula grafts required 11 re-operations, the majority for infections and non-union. Technical details of the operative procedure are reviewed for their part in the complications. PMID- 8220853 TI - The anatomic basis and clinical applications of flaps based on the posterior tibial vessels. AB - The vascular anatomy of the posterior tibial vessels has been studied in 20 legs of 10 cadavers. The number, size and distribution of the direct cutaneous and direct muscle branches of the posterior tibial vessels were recorded. For analysis, the leg was divided into four equal segments, Zone I being the most distal and Zone IV the most proximal. The direct cutaneous branches were found to cluster mainly around Zone II. The direct muscle branches to the soleus and flexor digitorum longus muscles arose chiefly in Zones II and III. Based on this knowledge, we performed fasciocutaneous, island soleus muscle, musculo fasciocutaneous and flexor digitorum longus muscle flaps based on the posterior tibial vessels in 12 patients. Four were free fasciocutaneous flaps and eight were pedicled flaps. All flaps survived and there were no major complications. Delayed wound healing was encountered in three patients. PMID- 8220854 TI - A distally based island first dorsal metatarsal artery flap for the coverage of a distal plantar defect. AB - An intractable plantar ulcer of the great toe was successfully covered with a distally based island first dorsal metatarsal artery flap. Common dorsal digital veins served as venous drainage for the flap. The results show that this flap can be used as a reliable alternative to conventional techniques to repair a distal foot defect. PMID- 8220855 TI - Innervated reverse digital artery flap through bilateral neurorrhaphy for pulp defects. AB - To provide sensation to the reverse digital artery (RDA) flap, both the dorsal branch from the proper digital nerve and the superficial sensory branch from the corresponding radial or ulnar nerve are sectioned at their proximal ends and included with the RDA flap. These are then anastomosed with the distal ends of both radial and ulnar digital nerves at the recipient wound. Three cases of pulp defects reconstructed by this technique achieved very favourable functional and cosmetic results. The RDA flap, innervated through bilateral neurorrhaphy, seems to be an excellent option for one-stage reconstruction of major pulp defects. PMID- 8220856 TI - Neurocutaneous axial island flaps in the forearm: anatomical, experimental and preliminary clinical results. AB - The neurocutaneous island flap is an axial flap composed of one vein and one nerve, the arterial vascularisation of which is provided by the vascular plexus around and inside the nerve. The vascularisation of the lateral and medial cutaneous nerve of the forearm was studied in twenty fresh human upper limbs. In ten rats the vascularisation of these nerves was studied using latex injection and in twenty other animals neurocutaneous island flaps were raised based on the lateral forelimb cutaneous nerve and the medial cutaneous nerves of the forelimb. There was always a consistent arterial longitudinal plexus along the human and animal nerves, which linked the cutaneous perforating arteries. All the experimental flaps harvested survived well. On the basis of these anatomical and experimental findings neurocutaneous island flaps were raised on the forearms of three patients and are reported on. PMID- 8220857 TI - Basal encephalocoele: imaging and exposing the hernia. AB - Basal encephalocoeles are rarely reported anomalies. Eight cases seen by one unit manifested external facial features and internal cerebral anomalies characteristic of the individual encephalocoele subgroups. CT and MR imaging delineates the anatomy of the skeletal defect and the associated cerebral abnormalities. Such imaging of cases of median cleft face syndrome may identify previously unsuspected basal encephalocoeles. Transcranial correction with increased exposure, if needed, by the technique of facial bipartition has been performed in five cases. PMID- 8220858 TI - The galea frontalis myofascial flap in anterior fossa CSF leaks. AB - We report the clinical use of galea frontalis myofascial flaps in the treatment of anterior fossa cerebrospinal fluid leaks after trauma. This flap provides an adequately sized and vascularised barrier between the cranial and nasal cavities through which the cells of the inflammatory response reach the target area. This technique was used in 9 cases with complete success; in 5 out of 9 patients, repair of an anterior cranial base bone defect was also performed with split calvarial bone grafts, harvested from the frontal craniotomy bone. In all patients, neither recurrence of the CSF leakage nor postoperative meningitis or its recurrence were observed. PMID- 8220859 TI - The scapular fasciocutaneous flap: a new flap for reconstruction of the posterior neck. AB - The axial, myocutaneous and free flaps have made immediate reconstruction of head and neck defects possible. Notwithstanding this remarkable progress, defects of the posterior neck leave the reconstructive surgeon with very little choice. The scapular fasciocutaneous flap is easy to harvest, reliable and versatile, with functional and cosmetic results comparable to free flaps. We describe the anatomy and the technique of this new donor site, along with a case report. PMID- 8220860 TI - Early postoperative brachytherapy following free flap reconstruction. AB - Brachytherapy delivered within the early postoperative period has been associated with delayed wound healing and wound breakdown. The objective of this study was to determine whether reconstruction with a microvascular free flap reduced the incidence of wound breakdown in the presence of early postoperative brachytherapy following wide excision of soft tissue sarcomas and head and neck carcinomas. Ten patients with malignant tumours underwent wide excision and free flap reconstruction. Brachytherapy was administered using Iridium-192 wires in the early postoperative period via tubes inserted intra-operatively. In 9 of the 10 patients the wounds healed uneventfully, demonstrating that brachytherapy can be delivered in the early postoperative period following free flap reconstruction without an increase in the frequency of wound breakdown. PMID- 8220861 TI - Early experience with the Lejour vertical scar reduction mammaplasty technique. AB - Over a 19-month period 25 bilateral reduction mammaplasties were performed using the Lejour technique. Results were assessed retrospectively. The mean follow-up was 12 months. The study showed that minor complications were common (40%), that nipple-areolar sensation was frequently reduced and that minor revision surgery for persistent dog-ears was often necessary (20%). Comparison with complications and revision rates in a similar group of patients who had undergone reduction mammaplasty using an inferior pedicle technique showed remarkably similar results. The positive findings of the current study were that nipple-areola necrosis was not encountered, patient satisfaction was high and the aesthetic results were generally good. PMID- 8220862 TI - Bupivacaine and Kaltostat reduces post-operative donor site pain. AB - A prospective double blind controlled trial was carried out to examine the differences in post-operative split skin graft donor site pain between sites dressed with three differently treated types of dressing; a dry calcium alginate dressing (Kaltostat Britcair), a saline moistened Kaltostat dressing and a bupivacaine hydrochloride (0.5%) moistened Kaltostat dressing. There was a significant reduction in post-operative pain in the Kaltostat and bupivacaine group (group 3) at 24 and 48 h when compared to the other two groups (p < 0.04). There was no difference in ease of removal of dressings or the quality of wound healing on day 10 between the three groups. This study demonstrates a significant reduction in post-operative pain in bupivacaine soaked Kaltostat without reducing the beneficial effects of Kaltostat on donor site healing and we recommend its use in clinical practice. PMID- 8220863 TI - Malignant melanoma excision margins: plastic surgery audit in Britain and Ireland, 1991, and a review. AB - In 1991, 146 consultant plastic surgeons in Britain and Ireland were sent a short questionnaire about their policies for excision margins for primary cutaneous malignant melanoma. 106/146 (73%) replied. 39/106 (37%) considered narrow margin (2-3 mm) excision biopsies adequate in certain cases. The minimum tumour thickness for a margin of more than 1 cm was 1 mm or more for 67/106 (63%) on the leg and for 57/106 (54%) on the back. The maximum margin was specified as 4 cm or more on the leg by 37/106 (35%) and on the back by 42/106 (40%). Other sub-groups of results were analysed. A review of the literature is presented. PMID- 8220864 TI - Human orf. AB - Human Orf is an uncommon viral disease acquired through contact with infected sheep and goats. We report five cases of human Orf acquired while preparing mutton. The clinical picture and the management of human Orf are presented. Awareness of the benign nature of the condition is important in preventing ill advised therapy. PMID- 8220865 TI - Identical unusual subtotal penile amputation in children: a report of four cases. AB - 4 cases are reported in whom an accidental or iatrogenic near complete slow amputation of the penis at the corona glandis was observed at first presentation. A representative case is presented in detail with illustrations showing the surgical repair carried out. The future erectile ability of this reconstructed phallus is speculated upon. PMID- 8220866 TI - Pressure developed under pressure garment. AB - To study how much pressure develops under a pressure garment, and how to obtain adequate effective pressure for scar treatment, we have used garments in combination with sponge and plastic plates in human volunteers. The pressures increased significantly when a sponge or sponge and plastic plate were used beneath the garment compared to pressures using a garment alone. In evaluations in patients with scar tissue on the limb, similar findings resulted. PMID- 8220867 TI - Apert's Syndrome. PMID- 8220868 TI - First use of tongue flap. PMID- 8220869 TI - The case of the disappearing leech. PMID- 8220870 TI - The effect of platelet-activating factor on the responsiveness of the human nasal airway. AB - 1. The effects of inhaled platelet-activating factor (PAF) on responsiveness of the human nasal airway were examined in normal subjects by measuring nasal airway resistance in response to histamine and bradykinin at 2, 6, 24, 48 h and 7 d after PAF administration. Eosinophil cationic protein (ECP) in nasal secretions was also measured. 2. Intranasal aerosol administration of PAF, 30 or 60 micrograms per nostril to normal human subjects induced an increased responsiveness to inhaled histamine, 50 to 400 micrograms and bradykinin, 100 micrograms per nostril at 2, 6 and 24 h following PAF treatment. However the effect was not apparent at 48 h or 7 days after PAF administration. 3. Intranasal administration of lyso-PAF, 60 micrograms by aerosol did not increase the reactivity of the nasal airway in response to histamine, 200 micrograms. 4. There was no difference in the time course of the PAF-induced hyperresponsiveness to histamine or bradykinin. 5. PAF-induced nasal hyperresponsiveness at 2 and 6 h was associated with increases in the ECP concentration of the nasal lavage fluid. 6. Vitamin E pretreatment of subjects resulted in the attenuation of the PAF induced hyperresponsiveness to histamine, and a decrease in ECP levels of the nasal lavage fluid. 7. The results suggest that in the human nasal airway, PAF induces a non-specific hyperresponsiveness which is accompanied by eosinophil activation in the nasal cavity. Free radical production induced by PAF may contribute to the hyperresponsiveness and the activation of eosinophils. PMID- 8220871 TI - RS 23597-190: a potent and selective 5-HT4 receptor antagonist. AB - 1. The pharmacological properties of RS 23597-190 (3-(piperdine-1-yl)-propyl-4 amino-5-chloro-2-methoxy benzoate hydrochloride) have been studied in vitro and in vivo. 2. RS 23597-190 competitively antagonized 5-HT4 receptor-mediated relaxations of rat, carbachol precontracted oesophageal muscularis mucosae, (pA2 = 7.8 +/- 0.1; Schild slope = 1.2 +/- 0.2). Affinity estimates (-log KB) at 5-HT4 receptors using either renzapride or SC-53116 as agonists yielded a -log KB value of 8.0 +/- 0.01. In contrast, RS 23597-190 failed to antagonize contractile responses to 5-HT of guinea-pig ileal 5-HT3 receptors, even at concentrations up to 10 microM. 3. Increases in short-circuit current, induced by 5-HT, were studied in guinea-pig ileal mucosal sheets. Concentration-response curves to 5-HT were biphasic, with the high potency phase to 5-HT inhibited by RS 23597-190 and mimicked by 5-methoxytryptamine. The -log KB value for RS 23597-190 at the high potency phase was 7.3 confirming that 5-HT4 receptors mediated the high potency phase. 4. In rat isolated vagus nerve, 5-HT elicited a slow, maintained depolarization at low concentrations and a rapid, transient depolarization at higher concentrations. The high potency, slow depolarizing phase to 5-HT was abolished selectively in the presence of 1 microM RS 23597-190 and the low potency phase was abolished selectively in the presence of 1 microM ondansetron. These data confirm that 5-HT4 and 5-HT3 receptors mediated slow and fast depolarization responses, respectively. 5. At 5-HT3 binding sites in membranes from NG 108-15 cells, labelled by [3H]-quipazine, RS 23597-190 exhibited an apparent affinity (- log Ki) of 5.7 +/- 0.1. At 5-HT3 receptors in membranes from rat cerebral cortex, labelled by [3H]-RS 42358-197, the apparent affinity (- log Ki) of RS 23597-190 was also 5.7 +/- 0.1. In both studies, Hill coefficients were not significantly different from unity. At 5-HT1A, 5-HT2,muscarinic M1, M2, M3, M4 and dopamine D1 and D2 receptors, RS 23597-190 exhibited low apparent affinities, with all - log Ki values less than 5.5.6. Intravenous infusion of RS 23597-190 in the conscious, restrained rat antagonized the von Bezold Jarisch reflex induced by 2-methyl 5-HT, with an ID50 of 300 microg kg-1 min-1, i.v. In the anaesthetized,bilaterally vagotomized micropig, RS 23597-190 (6 mg kg-1, i.v.) antagonized 5-HT-induced tachycardia with a half-life of 77 (63-99) min. Transient arrhythmic effects were noted after administration of the compound.7. In conclusion, RS 23597-190 acts as a high affinity, selective competitive antagonist at 5-HT4 receptors. Thus, the compound appears to be a useful tool for 5-HT4 receptor identification in vitro. In vivo, the compound is rapidly metabolized in pigs such that 5-HT4 blockade is not maintained. However,in the rat, when given by infusion, RS 23597-190 antagonizes 5-HT3 mediated responses, at doses consistent with a low affinity 5-HT3 receptor. These data suggest that, under appropriate experimental conditions, RS 23597-190 may also be used in vivo to characterize further 5-HT4 receptor function. PMID- 8220872 TI - Leukotriene D4- and prostaglandin F2 alpha-induced airflow obstruction and airway plasma exudation in guinea-pig: role of thromboxane and its receptor. AB - 1. We studied the effects of a thromboxane A2 receptor (TP receptor) antagonist, ICI-192,605 (0.5 mg kg-1, i.v.) and a selective thromboxane (Tx) synthetase inhibitor, OKY-046 (30 mg kg-1, i.v.), on airway responses induced by leukotriene D4 (LTD4; 0.2 nmol) or prostaglandin F2 alpha (PGF2 alpha; 20 nmol) instilled via the airways route to anaesthetized guinea-pigs. For a comparison, airway responses to a TxA2-mimetic, U-46619 (0.02 nmol) were also studied. We measured both lung resistance (RL) to monitor airflow obstruction, and extravasation of Evans Blue dye to quantify airway plasma exudation. 2. Instilled LTD4 into the tracheal lumen induced an immediate peak and subsequently persistent increase in RL and produced a large amount of extravasation of Evans Blue dye at all airway levels. Both ICI-192,605 and OKY-046 significantly attenuated the persistent increase in RL following the immediate response and reduced LTD4-induced extravasation of Evans Blue dye in the trachea and proximal intrapulmonary airway. Instilled LTD4 produced significant increases in immunoreactive TxB2 in bronchoalveolar lavage fluid obtained 1.5 min after instillation of LTD4. 3. Instilled PGF2 alpha into the tracheal lumen induced an immediate increase in RL which peaked at approximately 15 s. We also observed a delayed sustained increase in RL, reaching a second peak at approximately 4 min. PGF2 alpha produced small but significant increases in the amount of Evans Blue dye at all airway levels. As with PGF2 alpha, instillation of U-46619 produced a biphasic increase in RL and extravasation of Evans Blue dye. The potency of PGF2a, in inducing these airway responses was about 1000 times less than U-46619. ICI-192,605 abolished both the immediate and the delayed increase in RL after PGF2a, and also blocked PGF2a,-induced extravasation of Evans Blue dye. However, OKY-046 had no inhibitory effects on these responses.4. We conclude that airflow obstruction and airway plasma exudation induced by instilled LTD4 is, in part, mediated via TxA2 generation and subsequent activation of TP-receptors. On the other hand,instilled PGF2a, while inducing similar responses, does so primarily by direct activation of TP receptors,rather than via TxA2 generation. PMID- 8220873 TI - Characterization of P2-purinoceptor mediated cyclic AMP formation in mouse C2C12 myotubes. AB - 1. The formation of adenosine 3':5'-cyclic monophosphate (cyclic AMP) and inositol(1,4,5)trisphosphate (Ins(1,4,5)P3), induced by ATP and other nucleotides was investigated in mouse C2C12 myotubes. 2. ATP (100 microM) and ATP gamma S (100 microM) caused a sustained increase in cyclic AMP content of the cells, reaching a maximum after 10 min. The cyclic AMP content reached a maximum in the presence of 100 microM ATP, followed by a decline at higher ATP concentrations. ATP-induced cyclic AMP formation was inhibited by the P2-purinoceptor antagonist, suramin. 3. Myotubes hydrolysed ATP to ADP at a rate of 9.7 +/- 1.0 nmol mg-1 protein min-1. However, further hydrolysis of ADP to AMP and adenosine was negligible. 4. The cyclic AMP formation induced by ADP (10 microM-1 mM) showed similar characteristics to that induced by ATP, but a less pronounced decline was observed than with ATP. ADP-induced cyclic AMP formation was blocked by suramin, while cyclic AMP formation elicited by adenosine (10 microM-1 mM) was insensitive to suramin. 5. The ATP analogue, alpha,beta-methylene-ATP also induced a suramin sensitive cyclic AMP formation, while 2-methylthio-ATP and the pyrimidine, UTP, did not affect cyclic AMP levels. 6. Stimulation of the myotubes with ATP or UTP (10 microM-1 mM) caused a concentration-dependent increase in the Ins(1,4,5)P3 content of the cells. ADP (100 microM-1 mM) was less effective. Adenosine did not affect Ins(1,4,5)P3 levels. 7. Incubation of the cells with UTP (30 microM- 1 mM) inhibited the ATP- and ADP-induced cyclic AMP formation, suggesting that stimulation of the 'nucleotide' type P2-receptor inhibits P2-purinoceptor mediated cyclic AMP formation in C2C12 myotubes. In contrast, UTP (30 microM-I mM) enhanced adenosine-induced cyclic AMP formation.8. Adenosine-sensitive P1 purinoceptors activating cyclic AMP formation were found in C2C12 myotubes.Further, a novel P2-purinoceptor is postulated, sensitive to ATP, ADP and ATPgammaS, which also activates the formation of cyclic AMP in C2C12 myotubes. PMID- 8220874 TI - Partial inhibition by epithelium of tracheal smooth muscle relaxation induced by the potassium channel activator, BRL 38227. AB - 1. A method is described whereby either the serosal (Out) or epithelial (In) sides of rat isolated tracheae were selectively perfused. Perfusion with BRL 38227 (10(-8)-5 x 10(-6) M; In/Out) of preparations with intact epithelium (+ EP) precontracted with carbachol (10(-6) M; Out/In) produced complete relaxation. Perfusion with aminophylline (10(-5)-10(-3) M; In) of + EP preparations precontracted with carbachol (10(-6) M; Out) also produced complete relaxation. 2. In preparations precontracted with carbachol (10(-6) M) epithelium removal (- EP) increased the sensitivity to the relaxant effect of BRL 38227 (In), but not BRL 38227 (Out) [- log EC50, + EP/- EP; carbachol (In), BRL 38227 (Out): 6.76 +/- 0.11 vs 6.67 +/- 0.15; carbachol (Out), BRL 38227 (In): 5.93 +/- 0.06 vs 6.25 +/- 0.07]. Removal of the epithelium increased also the sensitivity to BRL 38227 (In) of preparations precontracted with a lower concentration (5 x 10(-7) M) of carbachol (Out). [- log EC50, + EP/- EP, carbachol (Out), BRL 38227 (In): 6.19 +/ 0.14 vs 6.58 +/- 0.17]. 3. Removal of the epithelium did not affect the sensitivity to BRL 38227 (In) of preparations precontracted with a higher concentration (5 x 10(-6) M) of carbachol (Out). 4. In both + EP and - EP preparations precontracted with carbachol (10(-6) M; Out), BRL 38227 (In) had a more potent relaxant effect than aminophylline (In) (EC50, BRL 38227 vs aminophylline, + EP/- EP: 5.93 +/- 0.06 vs 3.66 +/- 0.11/6.25 +/- 0.07 vs 3.77 +/ 0.11). 5. In preparations precontracted with carbachol (10-6 M; Out), removal of the epithelium did not affect the sensitivity to aminophylline (In) but increased the degree of precontraction (Tmax) following epithelial but not serosal stimulation with carbachol.6. We conclude that BRL 38227, a K+ channel activator, is a potent relaxant of rat tracheal smooth muscle precontracted with carbachol, and that the effect can be partially inhibited by the presence of an intact tracheal epithelium, whereas the relaxant effect of aminophylline is not. PMID- 8220875 TI - Kinetics of rate-dependent slowing of intraventricular conduction by the class Ib antiarrhythmic agent tocainide in vivo. AB - 1. The effects of the class I antiarrhythmic agent, tocainide, on intraventricular conduction were assessed in guinea-pigs, anaesthetized with pentobarbitone sodium 60 mg kg-1, i.p. 2. After electrical ablation of the sinus node, heart rate was controlled by atrial pacing. His bundle electrograms were recorded by means of an epicardial bipolar electrode. 3. During continuous stimulation, comparison of HV intervals measured at a cycle length of 475 ms, with HV intervals measured at a cycle length of 250 ms yielded the following results: 25.26 +/- 0.64 ms versus 25.02 +/- 0.70 ms (NS), at baseline, 26.65 +/- 0.80 ms versus 29.88 +/- 1.13 ms (P < 0.001) after i.v. administration of 30 mg kg-1 tocainide, and 28.04 +/- 0.64 ms versus 36.24 +/- 1.31 ms (P < 0.001), after addition of 20 mg kg-1 tocainide. Thus, tocainide caused HV intervals to increase in a strictly rate-dependent fashion. 4. In order to characterize the rate dependent class I activity of tocainide in terms of its binding kinetics to sodium channels, fractional sodium channel block was estimated from drug induced reductions of intraventricular conduction velocity (delta theta). On abruptly changing the drive cycle length from 500 ms to 250 ms, delta theta reached a new steady state with rate constants of 1.23 +/- 0.09 beat-1 and 1.28 +/- 0.09 beat 1, after administration of 30 mg kg-1 and addition of 20 mg kg-1 tocainide, respectively. At a basic drive cycle length of 250 ms delta theta recovered with time constants of 250.29 +/- 23.32 ms and 183.04 +/- 8.03 ms after administration of 30 mg kg-1 and addition of 20 mg kg-1 tocainide, respectively.5. The experimentally determined kinetic parameters were implemented into a mathematical model that assumes drug binding to sodium channels in terms of a periodical two state process. Rate-dependent reductions in conduction velocity during continuous stimulation after administration of tocainide were closely approximated by steady state reductions in sodium channel availability as calculated on the basis of the aforementioned model.6. In agreement with previously published in vitro studies, our data, obtained in vivo, confirm the classification of tocainide as a class I antiarrhythmic agent with fast onset and offset kinetics. The kinetic parameters obtained in vivo can be used in order to predict steady state reductions in conduction velocity at a wide range of frequencies. PMID- 8220876 TI - Regional haemodynamic effects of angiotensin II (3-8) in conscious rats. AB - 1. It has been reported that angiotensin II (AII) (3-8) causes endothelium dependent renal cortical vasodilatation, in anaesthetized rats, through interaction with a novel receptor that shows no affinity for the AT1-receptor antagonist, losartan. Therefore in order to get a fuller profile of the regional haemodynamic effects of AII (3-8) in conscious rats we assessed its renal, mesenteric and hindquarters vascular effects, and compared them to the responses elicited by AII and AIII. 2. AII and AIII (1.25, 12.5 and 125 pmol kg-1) caused dose-dependent pressor and renal and mesenteric vasoconstrictor effects. At doses up to 125 pmol kg-1, AII (3-8) was without any cardiovascular effects, but with doses of 1.25 and 12.5 nmol kg-1 there were dose-dependent increases in mean arterial blood pressure and reductions in renal and mesenteric flows and vascular conductances. The responses to AII (3-8) (12.5 nmol kg-1) were abolished by losartan (20 mumol kg-1). 3. Since it has been found that pretreatment with L arginine can reveal a vasodilator effect of AII (3-8) on rabbit pial arterioles, we assessed responses to AII (3-8) (12.5 nmol kg-1) before and 5 min after onset of a primed infusion of L-arginine (1.4 mmol kg-1 bolus, 1.4 mmol kg-1 h-1 infusion). Responses to AII (3-8) were unchanged by L-arginine. 4. The results are consistent with AII (3-8) being a less effective agonist than All (or AIII) at the AT1-receptor, but provide no evidence for AII (3-8) interacting with a novel receptor that shows no affinity for losartan. PMID- 8220877 TI - Effects of the central analgesic tramadol and its main metabolite, O desmethyltramadol, on rat locus coeruleus neurones. AB - 1. Tramadol is a centrally acting analgesic with low opioid receptor affinity and, therefore, presumably additional mechanisms of analgesic action. Tramadol and its main metabolite O-desmethyltramadol were tested on rat central noradrenergic neurones of the nucleus locus coeruleus (LC), which are involved in the modulation of nociceptive afferent stimuli. 2. In pontine slices of the rat brain the spontaneous discharge of action potentials of LC cells was recorded extracellularly. (-)-Tramadol (0.1-100 microM), (+)-tramadol (0.1-100 microM), ( )-O-desmethyl-tramadol (0.1-100 microM) and (+)-O-desmethyltramadol (0.01-1 microM) inhibited the firing rate in a concentration-dependent manner. (+)-O desmethyltramadol had the highest potency, while all other agonists were active at a similar range of concentrations. 3. (-)-Tramadol (10, 100 microM) was less inhibitory in brain slices of rats pretreated with reserpine (5 mg kg-1, 5 h before decapitation) than in controls. 4. The effect of (-)-tramadol (10 microM) was abolished in the presence of the alpha 2-adrenoceptor antagonist, rauwolscine (1 microM), whilst that of (+)-O-desmethyltramadol (0.3 microM) virtually disappeared in the presence of the opioid antagonist, naloxone (0.1 microM). (+) Tramadol (30 microM) and (-)-O-desmethyl-tramadol (10 microM) became inactive only in the combined presence of naloxone (0.1 microM) and rauwolscine (1 microM). 5. In another series of experiments, the membrane potential of LC neurones was determined with intracellular microelectrodes. (-)-Tramadol (100 microM) inhibited the spontaneous firing and hyper-polarized the cells; this effect was abolished by rauwolscine (1 microM). (+)-O-desmethyltramadol (10 microM)had a similar but somewhat larger effect on the membrane potential than ( )-tramadol. The (+)-O-desmethyltramadol-(10 microM) induced hyperpolarization was abolished by naloxone (0.1 microM).6. The hyperpolarizing effect of noradrenaline (30 microM) was potentiated in the presence of (-)-tramadol(100 microM), but not in the presence of (+)-O-desmethyltramadol (10 microM). There was no potentiation of the noradrenaline (30 microM) effect, when the cells were hyperpolarized by current injection to an extent similar to that produced by (-)-tramadol (100 microM).7. Both noradrenaline (100 microM) and (- )-tramadol (100 microM) decreased the input resistance.8. The results confirm that the analgesic action of tramadol involves both opioid and non-opioid components. It appears that (-) tramadol inhibits the uptake of noradrenaline and via a subsequent increase in the concentration of endogenous noradrenaline indirectly stimulates alpha2 adrenoceptors. (+)-0-desmethyltramadol seems to stimulate directly opioid micro receptors. The effects of (+)-tramadol and(-)-O-desmethyltramadol consist of combined micro-opioid and alpha2-adrenergic components. PMID- 8220878 TI - Use-dependent block of Na+ currents by mexiletine at the single channel level in guinea-pig ventricular myocytes. AB - 1. The mechanism of use-dependent block of Na+ current by mexiletine was studied at the single channel level in guinea-pig ventricular myocytes by the patch-clamp techniques. All experiments were performed using stimulation protocols to enable us to analyze the strict dependence of changes in channel properties on channel use. 2. In cell-attached patches, bath or pipette application of mexiletine (40 microM) produced a use-dependent reduction of the peak average current without changes in single channel conductance. Null sweeps were increased and the number of openings per sweep decreased with successive pulses, whereas no significant change in the mean open time was detected during the train. 3. Block by mexiletine became greater when pulse duration was extended beyond the period in which channels were open, suggesting that block progressed without channel opening. 4. At near threshold potentials, mexiletine decreased the later occurrence of first openings. Additionally, late openings were reduced in a use dependent way. 5. We conclude that mexiletine binds to the inactivated closed states of the Na+ channel and then causes a failure of late openings as well as early, which results in null sweeps on subsequent depolarization. PMID- 8220879 TI - Evidence for participation of B1 and B2 kinin receptors in formalin-induced nociceptive response in the mouse. AB - 1. This study was designed to investigate the role of bradykinin (BK), as well as the subtype of BK receptors involved, in formalin-induced hindpaw pain in the mouse by use of selective B1 and B2 receptor antagonists. In addition, we have analysed whether or not BK may be involved in formalin-induced hindpaw oedema in the mouse. 2. The pretreatment of animals with captopril (2 and 5 mg kg-1, s.c.) significantly increase the first and the second phases of formalin-induced pain. 3. Co-injection of the selective B1 receptor antagonist des-Arg9[Leu8]-BK (0.2 0.4 nmol/paw), together with formalin, caused graded and similar inhibitions of both phases of formalin-induced pain. Similar results were obtained with the B2 antagonists NPC 349 (D-Arg[Hyp3,Thi5,8-D-Phe7]-BK) and NPC 567 (D-Arg[Hyp3, D Phe7]-BK) (0.2 and 0.6 nmol/paw). Higher concentrations of these antagonists (1 nmol/paw) failed to antagonize formalin-induced pain. 4. The new potent and selective B2 receptor antagonists, Hoe 140 (D-Arg[Hyp3,Thi5,D-Tic7,Oic8]-BK), NPC 17731 (D-Arg[Hyp3, trans-4-propoxy-D-proline (transpropyl)7, Oic8]-BK), and NPC 17761 (D-Arg[Hyp3, trans-4-propoxy-D-proline (trans thiophenyl)7, Oic8]-BK) (0.02 to 1.0 nmol/paw), also caused significant inhibitions of both phases of formalin induced pain. When Hoe 140 was injected subcutaneously 30 min before formalin injection (9.9 and 99 nmol kg-1), it significantly attenuated both phases of formalin-induced pain. The putative non-peptide BK antagonist, MV 8612 (1.6 to 9.6 nmol/paw), but not MV 8608 (5.5 to 33 nmol/paw), caused a graded inhibition of both phases of formalin induced pain, being, however, more active against the first phase.5. The pretreatment of animals with morphine (2.6 to 13 micromol kg 1, s.c.) caused dose-dependent and equipotent inhibitions of both phases of formalin-induced pain. In contrast, in domethacin (2.7 to 27 micromol kg-1) antagonized only the second phase of formalin-induced pain.6. The B2 receptor antagonists, Hoe 140, NPC 17731, NPC 17761, NPC 349 and NPC 567, all caused a significant inhibition of formalin-induced hindpaw oedema. A similar inhibition was also observed within domethacin but not with captopril or morphine.7. Our results provide strong evidence for the important role of endogenous BK, acting through both B1 and B2 receptors, in the genesis of both phases of formalin induced persistent pain in the mouse. In addition, the current results also demonstrate that the inflammatory oedema associated with the later phase of formalin-induced pain seems to be mediated by endogenous BK, via activation of B2 receptors. PMID- 8220880 TI - Effects of hydroxyethylrutosides on the permeability of microvessels in the frog mesentery. AB - 1. We have investigated the effects of a standardised mixture of hydroxyethylrutosides (HR, Venoruton), a mixture of five of its main components (M) and each of the five components separately (7-mono-HR, 7,4'-di-HR, 7,3',4' tri-HR, 5,7,3',4'-tetra-HR and 7,3'4'-tri HQ) upon the permeability of single perfused capillaries and venules in the mesenteries of pithed frogs. 2. In each experiment, the hydraulic permeability (Lp) of a single perfused microvessel and the effective osmotic pressure (sigma delta pi) exerted by macromolecules across its walls were estimated by a microcclusion technique, first during control perfusion and then in the presence of a known concentration of test substance. 3. HR, M and 7,4'-di-HR reduced Lp in a similar concentration-dependent manner over the range of 1 microgram ml-1 to 1 mg ml-1 (maximum reduction was to 40% of control Lp at 1 mg ml-1). At perfusate concentrations greater than 1 mg ml-1, these substances reduced Lp to a lesser extent. While the four other test substances reduced Lp significantly when their perfusate concentrations equalled or exceeded 100 micrograms ml-1, they were all less potent than 7,4'-di-HR. 4. The reduction in Lp induced by the mixture of flavonoids was only slightly reversed by subsequent perfusion with flavonoid-free solutions. 5. When permeability was increased by perfusing with protein-free solutions, both HR and 7,4'-di-HR reduced and then reversed the increase in Lp in a concentration dependent manner over the range of 1 microgram ml-1 to 100 micrograms ml-1. None of the other component flavonoids was effective in restoring Lp under these conditions. PMID- 8220881 TI - Effects of L- and D-arginine and some related esters on the cytosolic mechanisms of alpha-thrombin-induced human platelet activation. AB - 1. In Fura-2 preloaded human platelets, the increase in cytosolic calcium induced by alpha-thrombin was reduced by some L- and D-arginine ester compounds the IC50 (microM) values of which were 7.4 for TAEE, 56.9 for BAEE, 77.6 for TAME, 560 for T(d)AME, 656.3 for L-ArgOMe and 2206.7 for D-ArgOMe. alpha-tosyl-L-Arginine, L- and D-arginine were inactive. 2. The inhibitory activity of the L-arginine esters was not modified when platelets were pretreated with 100 microM N omega monomethyl-L-arginine. 3. The L-arginine esters did not increase cyclic GMP content in platelets either in the presence or absence of indomethacin and apyrase at rest and after alpha-thrombin stimulation. 4. The kinetic parameters of platelet Na+/H+ antiporter (amiloride-inhibitable, evaluated after cytosolic nigericin-induced acidification) were modified by L- and D-arginine esters, while the native amino acids were ineffective. 5. The inhibitory effects of the L- and D-arginine esters on platelet activation appear to be mainly due to their inhibitory effect on Na+/H+ antiporter. PMID- 8220882 TI - Electrophysiological effects of Org 7797 in the closed-chest anaesthetized dog. AB - 1. The intravenous electrophysiological effects of a new antifibrillatory agent, Org 7797, were studied in closed chest anaesthetized dogs. Effects of fast sodium and slow calcium-mediated action potentials were also examined in guinea-pig isolated papillary muscle. 2. The major effects of a known antifibrillatory dose of Org 7797 (0.5 mg kg-1) were a protracted slowing of AV nodal conduction (for at least 20 min) and prolongation of the AV nodal functional refractory period. Conduction in the atria and His-Purkinje system (reflected by the St-A and HV intervals) were not significantly modified whilst ventricular conduction (reflected by the QRS interval) and the ventricular functional refractory period were only transiently prolonged. No other electrophysiological changes were seen. 3. A higher dose of Org 7797 (1.5 mg kg-1) slowed conduction at all levels of the myocardium (as evidenced by increases in the St-A, AH, HV and QRS intervals), slightly shortened cardiac repolarization (as assessed from JTc) and decreased Wenckebach rate. Atrial refractory periods were increased whereas effects on ventricular refractory periods were modest. 4. Neither heart rate nor sinus node recovery time were modified by either dose of Org 7797. 5. Org 7797, at a concentration (20 microM) which reduced Vmax of fast sodium-mediated action potentials in isolated papillary muscle by 83%, did not modify Vmax of slow calcium-mediated action potentials. It prolonged duration of the latter but did not modify that of the former. However, the plateau phase of both the 'fast' and especially the 'slow' action potentials was prolonged. 6. It is concluded that the main electrophysiological effects of a known antifibrillatory dose of Org 7797 in dogs with normal cardiac function are seen at the level of the AV node, actions which are unlikely to be explained by calcium channel block. Higher doses display a class Ic profile. This preferential action on the AV node may contribute to the control of ventricular rate during atrial fibrillation in the absence of infra-nodal conduction disturbances.7. These results contrast with those previously obtained in infarcted dogs and might further suggest that myocardial infarction enhances the Class I action of Org 7797. PMID- 8220883 TI - Eicosanoid-dependence of responses of pre- but not postglomerular vessels to noradrenaline in rat isolated kidneys. AB - 1. We investigated the role of nitric oxide (NO) and of vasoactive eicosanoids in the control of renal vascular resistance (RVR) and glomerular filtration rate (GFR) and of their responses to noradrenaline (NA). This study was conducted in single-pass perfused, isolated kidney preparations of the rat. 2. NA (63, 110 and 160 nM) dose-dependently increased RVR and to a lesser degree GFR. 3. In baseline conditions, N omega-nitro-L-arginine methylester (L-NAME, 100 microM) increased GFR more than RVR, thus demonstrating a basal release of NO which predominates in postglomerular vessels. 4. In kidneys stimulated with NA, L-NAME potentiated the increases in RVR but not in GFR. Indomethacin (1.5, 150 nM and 15 microM) did not alter GFR but markedly and dose-dependently reduced the NA-induced increase in RVR. Similar results were obtained with GR 32191B (10 and 100 microM), a prostaglandin H2/thromboxane A2 (PGH2/TxA2) receptor antagonist. 5. Indomethacin (15 microM) suppressed the enhancing effects of L-NAME on RVR responses to NA but did not affect those on GFR. 6. It is concluded that the mechanisms of the response to NA differ among pre- and postglomerular vessels. In preglomerular vessels the vasoconstrictor action and the NO release depend upon the activation of PGH2/TxA2 receptors, while both are eicosanoid-independent in the postglomerular vessels. PMID- 8220884 TI - Estimation of partial agonist affinity by interaction with a full agonist: a direct operational model-fitting approach. AB - 1. The operational model of agonism (Black & Leff, 1983) has been extended to describe the interaction between a partial agonist and a full agonist at the same receptor. The derived equation explicitly describes the interaction and allows the affinity (and efficacy) of the partial agonist to be estimated by direct fitting of raw experimental agonist concentration-effect (E/[A]) curve data. 2. The model was used to analyse experimental E/[A] curve data generated for the interaction between pilocarpine (partial agonist) and carbachol (full agonist) at the M3-muscarinic receptor mediating contraction of the guinea-pig isolated trachea. Pilocarpine affinity estimates obtained by operational model-fitting were compared with those obtained by use of the null method (Stephenson, 1956). These analyses demonstrated that the two methods gave comparable results (mean pKB estimates were 5.79 and 5.86 for the operational model and null method respectively). 3. When multiple concentrations of partial agonist are used, simultaneous operational model-fitting of all the E/[A] curve data allows the competitive nature of the interaction to be studied. 4. We conclude that operational model-fitting is a valid and analytically simple alternative to the conventional null method of analysing full/partial agonist interactions. PMID- 8220885 TI - Pharmacological characterization of the novel nonpeptide angiotensin II receptor antagonist, BIBR 277. AB - 1. The pharmacological profile of BIBR 277, 4'-[(1,4'-dimethyl-2'-propyl[2,6'-bi 1H-benzimidazol]-1'-yl)methyl ]- [1,1'-biphenyl]-2-carboxylic acid, a novel, nonpeptide angiotensin II receptor antagonist has been investigated by use of receptor binding studies, enzymatic assays, functional in vitro assays in rabbit aorta as well as in vivo experiments in pithed, anaesthetized and conscious rats. 2. BIBR 277 potently interacted with rat AT1 receptors (Ki 3.7 nM). Competitive receptor interaction was shown by radioligand saturation experiments performed in the presence of BIBR 277. The failure to inhibit radioligand binding to AT2 sites demonstrates the selectivity of BIBR 277 for AT1 receptors. This is further substantiated by the findings that BIBR 277 neither interacted with other receptor systems investigated nor affected the activity of components of the human renin-angiotensin system, such as plasma renin or serum converting enzyme. 3. In rabbit aorta, BIBR 277 had no agonistic properties and was shown to be an insurmountable antagonist of angiotensin II-induced contractions (KB 0.33 nM). The antagonistic effect persisted even after several wash-out procedures. However, this interaction was not irreversible since the insurmountable antagonism was concentration-dependently reversed when BIBR 277 (0.1 microM) and the surmountable antagonist, losartan (0.1 and 1.0 microM) were incubated simultaneously. The specificity of BIBR 277 for the AT1 receptor was further substantiated in this preparation since micromolar concentrations of BIBR 277 neither affected potassium chloride and noradrenaline-induced contractions nor acetylcholine-mediated tissue relaxation. 4. In pithed rats, i.v. administration of BIBR 277 (0.1, 0.3 and 1.0 mg kg-1) shifted the dose-pressor response curve to angiotensin II dose-dependently to the right with ED50 values of 0.23 microg kg-1 (control)and 1.4 microg kg-1, 4.7 microg kg-1 and 20 microg kg-1, respectively. As observed in the in vitro experiments no agonistic effect was detected and the maximum of the blood pressure response to angiotensin II at the highest dose of BIBR 277 was decreased by 29%.5. In anaesthetized rats, bolus i.v. administration of 0.1, 0.3 and 1.0 mg kg-1 BIBR 277 attenuated the blood pressure response to bolus i.v. injections of angiotensin 11 (0.1 microg kg-1). At the highest dose an almost complete blockade was observed even after 2 h.6. Single oral administration of BIBR 277 (0.3 and 1.0 mg kg-1) to conscious, chronically instrumented renovascular hypertensive rats dose-dependently decreased the mean arterial blood pressure by 15 and 30 mmHg, respectively. At the higher dose a significant antihypertensive effect was maintained for more than 24 h. Moreover, consecutive daily dosing of 1 mg kg-1 orally resulted in a sustained reduction in blood pressure over the 4 day observation period.7. It is concluded that BIBR 277 is an effective and selective angiotensin II antagonist with antihypertensive activity after oral administration. PMID- 8220886 TI - Centrally administered ouabain aggravates rapid-eye-movement-sleep-related bradyarrhythmias in freely moving rats. AB - 1. The effects of continuous infusions of ouabain on bradyarrhythmias (cardiac pauses for 0.5 s or longer) during sleep were examined in freely moving Wistar Kyoto rats. 2. In a control group (n = 7), saline was infused into both the lateral ventricle and the femoral vein. In an intracerebroventricular (i.c.v.) ouabain group (n = 7), ouabain was infused centrally, such that each rat received three stepped doses of 1, 10, and 100 ng kg-1 h-1 for 3 days at each dose, while saline was infused systemically. In an intravenous (i.v.) ouabain group (n = 7), ouabain was infused systemically at the same doses as the i.c.v. ouabain received, while the simultaneous i.c.v. infusion of saline was carried out. 3. Three-day i.c.v. infusions of the three stepped doses of ouabain caused a dose dependent increase in the frequency of bradyarrhythmias during rapid-eye movement (REM) sleep without affecting the time spent in REM sleep, arterial pressure, average heart rate, or the frequency of bradyarrhythmias during non-REM sleep. Intravenous ouabain or i.c.v. saline had no effects on the frequency of bradyarrhythmias. 4. Intrinsic CNS activity during REM sleep may be involved in the centrally mediated arrhythmogenic properties of ouabain during sleep. PMID- 8220887 TI - Effects of morphine metabolites on micturition in normal, unanaesthetized rats. AB - 1. By means of continuous cystometry in normal, unanaesthetized rats, the effects on micturition of intrathecally (i.t.) administered morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G), the two main metabolites of morphine, were studied and compared with those of i.t. morphine. 2. Both M6G (0.01, 0.1, and 0.5 microgram) and M3G (5 micrograms) were found to have significant effects on micturition. Like morphine (0.1, 0.5, and 10 micrograms), M6G was able to inhibit the micturition reflex, and produce urinary retention and dribbling incontinence in a dose-dependent manner. The potency of M6G for inhibiting micturition was approximately 10 times higher than that of morphine, and the duration of its effect was longer. All effects of M6G could be reversed by naloxone. 3. M3G (5 micrograms) facilitated the micturition reflex, resulting in decreases in bladder capacity and micturition volume, and an increase in spontaneous contractile activity. Pretreatment with naloxone (10 micrograms), which by itself had no effect on micturition, enhanced the facilitatory effects of M3G. In addition, M3G tended to counteract the inhibitory effects of both morphine and M6G on micturition. M3G (5 micrograms) also produced an excitatory behavioural syndrome. 4. It is concluded that in rats, i.t. M3G has excitatory effects on micturition and behaviour, probably not mediated via opioid receptors. I.t M6G has a potent inhibitory effect on micturition mediated by stimulation of opioid receptors. It may have effects on somatosensory afferent input in lower doses than those required for effects on micturition. PMID- 8220888 TI - The effects of ATP and alpha,beta-methylene-ATP on cytosolic Ca2+ level and force in rat isolated aorta. AB - 1. The effects of a non-selective P2-receptor agonist ATP and a selective P2x receptor agonist alpha,beta-methylene-ATP on intracellular free Ca2+ level ([Ca2+]i) and force were examined in rat isolated aorta without endothelium. 2. Both ATP (1-1000 microM) and alpha,beta-methylene-ATP (0.1-100 microM) induced transient increase followed by small sustained increase in [Ca2+]i in a concentration-dependent manner. Compared with the force induced by a high concentration of KCl, the force induced by alpha,beta-methylene-ATP was smaller and that induced by ATP was much smaller at a given [Ca2+]i. 3. An L-type Ca2+ channel blocker, verapamil (10 microM), completely inhibited the high K(+) stimulated [Ca2+]i and force. Verapamil partially inhibited the transient and sustained increases in [Ca2+]i induced by 10 microM alpha,beta-methylene-ATP and the sustained increase but not the transient increase induced by 1 mM ATP. 4. In the absence of extracellular Ca2+ (with 0.5 mM EGTA) 1 mM ATP caused transient increase in [Ca2+]i while 10 microM alpha,beta-methylene-ATP was ineffective 5. ATP, but not alpha,beta-methylene-ATP, increased the tissue adenosine 3':5' cyclic monophosphate (cyclic AMP) level. 6. These data suggest that ATP and alpha,beta-methylene-ATP increase [Ca2+]i by an activation of both L-type and non L-type Ca2+ channels. In addition, ATP, but not alpha,beta-methylene-ATP, increases [Ca2+]i by a release of Ca2+ from an intracellular Ca2+ store. Possible reasons are discussed as to why the increase in [Ca2+]i due to ATP and alpha,beta methylene-ATP resulted in only a small contraction. PMID- 8220889 TI - Dissociation of the anti-ischaemic effects of cloricromene from its anti-platelet activity. AB - 1. Cloricromene is a non-anticoagulant coumarin derivative with anti-platelet and anti-leukocyte properties, which has beneficial effects in various models of ischaemia and shock. 2. We have assessed the effects of cloricromene on (a) ex vivo platelet aggregation, and (b) infarct size using a model of myocardial ischaemia in the anaesthetized rabbit. 3. Cloricromene (1-1000 micrograms kg-1 min-1 for 15 min) induced a dose-dependent inhibition of ex vivo platelet aggregation, causing only a minimal increase in heart rate and no change in mean arterial blood pressure. The inhibitory activity was considerably stronger when platelet aggregation was induced by collagen than by ADP. 4. Cloricromene inhibited ex vivo platelet aggregation in rabbits pretreated with indomethacin (5 mg kg-1) and this inhibition persisted for 30-60 min. 5. The model of myocardial ischaemia involved 1 h occlusion of the first antero-lateral branch of the left coronary artery followed by 2 h of reperfusion. Infusion of cloricromene (30 or 300 micrograms kg-1 min-1), ibuprofen (80 micrograms kg-1 min-1) or vehicle began 15 min prior to occlusion, and continued throughout the experiment. 6. While area at risk was similar for all groups studied, cloricromene (30 or 300 micrograms kg 1 min-1) or ibuprofen caused a reduction in infarct size, and decreased myeloperoxidase activity in the tissue of the infarcted myocardium. 7. Cloricromene at 300 micrograms kg-1 min-1 also reduced the occlusion-induced elevation of the ST-segment of the rabbit electrocardiogram, and inhibited platelet aggregation ex vivo. Ibuprofen or cloricromene at 30 fg kg-1 min-1 had no effect on either the ST-elevation or platelet reactivity.8. Thus, cloricromene exhibits a cardioprotective activity via an inhibition of leukocyte infiltration, in the presence (300 microg kg-l min-1) or absence (30 microg kg-1 min-1) of inhibition of platelet activity ex vivo.The anti-aggregatory activity of cloricromene acts via a mechanism that is either different from, or in addition to, inhibition of cyclo-oxygenase, and is of long duration. PMID- 8220890 TI - A facilitatory effect of anti-angiotensin drugs on vagal bradycardia in the pithed rat and guinea-pig. AB - 1. In pithed rats, preganglionic vagal nerve stimulation (at 5 Hz) elicited a bradycardia. This bradycardia was potentiated by the angiotensin converting enzyme inhibitor, captopril (1 mg kg-1, i.v.) by about 40%. Subsequent angiotensin II infusion (0.03 micrograms kg-1 min-1) reversed this effect. A similar facilitatory effect was also seen with the angiotensin receptor antagonist, losartan (10 mg kg-1, i.v.). These results suggest a tonic inhibitory effect of endogenous angiotensin II on vagal transmission. 2. The effect of captopril in potentiating vagal bradycardia appears to be at the level of vagal neurones, since the bradycardia elicited by the muscarinic agonist, methacholine was unaffected. 3. After the pithed rats were nephrectomized, captopril had no effect on vagally-induced bradycardia, suggesting that the formation of the endogenous angiotensin II responsible for the effect was dependent on renin release from the kidney. 4. When the sympathetic nerves of the pithed rat were electrically stimulated there was a tachycardia, and this was unaffected by captopril. However, when the sympathetic and vagus nerves were activated concurrently, the resulting tachycardia was inhibited by captopril. 5. In pithed guinea-pigs, captopril also potentiated the bradycardia caused by vagal nerve stimulation. This appears to be a tissue-selective effect since the bronchoconstriction due to the vagal stimulation was not affected by captopril. 6. These results suggest that endogenous angiotensin II can have a tonic inhibitory effect on cardiac vagal transmission. Disruption of this mechanism by anti-angiotensin drugs may attenuate the reflex tachycardia associated with the fall in blood pressure in anti-hypertensive therapy. PMID- 8220891 TI - Characteristics of the bradykinin-induced changes in intracellular calcium ion concentration of single bovine tracheal smooth muscle cells. AB - 1. Single bovine tracheal smooth muscle (BTSM) cells were cultured and used to measure bradykinin-induced changes in [Ca2+]i by dynamic video imaging. 2. Bradykinin (10 pM-10 microM)-induced an increase in [Ca2+]i over basal levels (69 +/- 2 nM; n = 353) which was concentration-dependent (log EC50 = -8.7 M) in the presence of extracellular calcium ions (2 mM). The bradykinin B2 receptor antagonist, D-Arg[Hyp3,Thi5,8,D-Phe7]- bradykinin, produced a parallel shift to the right of the bradykinin concentration-response curve (log EC50 = -7.1 M and 5.8 M in the presence of 1 microM and 10 microM antagonist respectively) yielding an apparent KD of 26 nM. 3. In the absence of extracellular calcium ions (with 0.1 mM EGTA), bradykinin (10 pM-10 microM) produced a uniform increase in [Ca2+]i from a basal level of 33 +/- 2 nM (n = 140) to approximately 180 nM in BTSM cells indicating an 'all-or-nothing' release of intracellular calcium ions. In the presence of 10 microM D-Arg[Hyp3,Thi5,8,D-Phe7]-bradykinin no responses could be induced by bradykinin at concentrations below 100 nM. However, at 100 nM and 1 microM bradykinin there was no change in the uniform increase in [Ca2+]i in these cells previously observed. 4. In both the absence or presence of D Arg[Hyp3,Thi5,8,D-Phe7]-bradykinin, there was a concentration-dependent increase in the percentage of cells responding to bradykinin (frequency) under calcium rich or calcium-free conditions. Individual cells also demonstrated a difference in the sensitivity to any particular concentration of bradykinin. 5. A latency in the response of cells to bradykinin was observed both in calcium-containing and calcium-free conditions.6. We conclude that bradykinin B2 receptors are expressed by BTSM cells and are involved in the bradykinin-induced increase in [Ca2+]i. It appears that the increase in [Ca2+], can be mediated via a graded influx of calcium ions from the extracellular space or an 'all-or-nothing' release from intracellular stores. PMID- 8220893 TI - Functional evidence for multiple receptor activation by kappa-ligands in the inhibition of spinal nociceptive reflexes in the rat. AB - 1. The evidence for kappa-receptor heterogeneity is equivocal. We have now investigated this question by comparing the effects of five putatively selective kappa-agonists. The parameters examined were: the relative potencies in depressing hindlimb flexor muscle reflexes to noxious pinch stimuli in both spinalized and sham-spinalized rats; the reversibility of these effects by naloxone; and the effects on blood pressure. 2. Two types of drug effect was discriminated. One drug group, represented by U-50,488, U-69,593 and PD-117,302, had a potency ratio between sham and spinalized rats approximately 10 fold lower than the other group, which comprised GR103545 and CI-977. 3. Under sham spinalized conditions, CI-977 and GR103545 at high doses caused only sub-maximal reductions of spinal reflexes. U-50,488 was still active when superimposed on these high doses of GR103545. 4. Naloxone reversed all effects, but different doses were required between compounds, with GR103545 taking some 20 times higher doses of naloxone to cause reversal than did U-50,488. 5. The effects on mean arterial pressure were opposite between groups. 6. The results imply that more than one type of naloxone-sensitive non-mu opioid receptor must be involved in mediating these complex actions of ligands that have been claimed to be selective for kappa-receptors. PMID- 8220894 TI - Post-receptor pathway of the ATP-induced relaxation in smooth muscle of the mouse vas deferens. AB - 1. The post-receptor pathway of the ATP relaxant effect in K(+)-precontracted vas deferens smooth muscle (VD) was examined. 2. The relaxation to ATP was not antagonized either by 10 microM methylene blue, a cyclic GMP inhibitor, by 10 microM indomethacin, an inhibitor of prostaglandin synthesis or by 100 microM NG nitro-L-arginine, an inhibitor of NO production. 3. The Rp-diastereomer of adenosine 3':5'-cyclic monophosphorothioate (Rp-cAMPS) 200 microM, a competitive inhibitor of cyclic AMP significantly diminished the relaxant response to ATP. 4. Isoprenaline 10 microM, a beta-adrenoceptor agonist, produced a sustained relaxation, inhibited by Rp-cAMPS, without a significant change in [Ca2+]i, thereby mimicking the ATP-induced relaxant effect. 5. The level of the phosphorylated myosin light chain in the precontracted VD was significantly lowered by 1000 microM ATP. 6. ATP (1000 microM) and isoprenaline (10 microM) produced the same increase (+ 50%) of [cyclic AMP] when applied to a resting VD. 7. The effect of simultaneous increases of [Ca2+]i and of [cyclic AMP] produced by externally applied ATP are discussed. 8. These results suggest that ATP induced relaxation in K(+)-precontracted VD is mediated by the activation of adenylyl cyclase. PMID- 8220895 TI - Evidence for sympathetic neurotransmission through presynaptic N-type calcium channels in human saphenous vein. AB - 1. The specific type(s) of voltage-sensitive calcium channels (VSCCs) involved in sympathetic neurotransmission have not yet been characterized in human vascular tissues. We therefore examined the functional role of the N- and L-type VSCCs in human saphenous veins. 2. Contractile response curves for transmural nerve stimulation (TNS) and for exogenously administered noradrenaline (NA) were obtained in superfused saphenous vein rings. The contractions induced by TNS, but not by NA, were inhibited by 1 microM tetrodotoxin and by 10 microM guanethidine. Both responses were substantially reduced by 1 microM phentolamine, indicating that the contractions evoked by TNS were mediated by endogenous NA released from noradrenergic nerves. 3. In the presence of 2 microM omega-conotoxin GVIA (omega Conus Geographus toxin, fraction VI A; omega-CgTx), a polypeptide with specific inhibitory activity on N- and L-type calcium channels, the neurally evoked contractions were almost completely abolished. In contrast, the responses induced by exogenous NA were not affected by the neurotoxin, thus providing evidence of the exclusive presynaptic action of omega-CgTx. 4. In the presence of the calcium antagonist verapamil (10 microM), which selectively blocks L-type VSCCs, the contractions induced by both TNS and NA were diminished to the same extent, suggesting that the organic calcium blocker is active only at the postjunctional level. 5. It is concluded that N-type calcium channels are the main pathway of calcium entry controlling the functional responses induced by activating sympathetic nerves; the role of L-type channels appears to be limited to the postjunctional level, modulating smooth muscle contractions. PMID- 8220892 TI - Thirteenth Gaddum Memorial Lecture. Neuronal and endothelium-derived mediators in the modulation of the gastric microcirculation: integrity in the balance. PMID- 8220896 TI - Effect of endopeptidase-24.11 inhibition and of atrial natriuretic peptide clearance receptor ligand on the response to rat brain natriuretic peptide in the conscious rat. AB - 1. The present studies examined the effect of (a) a specific endopeptidase-24.11 (E-24.11) inhibitor (candoxatrilat) and (b) a ligand for the atrial natriuretic peptide (ANP) clearance receptor (SC 46542) on the renal and blood pressure response to brain natriuretic peptide (BNP) in conscious rats. 2. Infusion of BNP 200 ng kg-1 min-1 for 60 min produced a small rise in urinary sodium and guanosine 3':5'-cyclic monophosphate (cyclic GMP) excretion with a non significant fall in mean arterial blood pressure. 3. Candoxatrilat (3 mg kg-1) alone had no significant effect on sodium excretion or blood pressure but markedly potentiated the natriuretic response to BNP. 4. Similarly SC 46542 (68 micrograms kg-1; 6.8 micrograms kg-1 min-1) which produced no significant effect on its own, potentiated the natriuresis-induced by BNP, although the effect was of shorter duration compared to that of candoxatrilat. 5. The data indicate two approaches to the potentiation of the renal activity of BNP and suggest that BNP may mediate some of the activity of E-24.11 inhibitors reported in cardiac failure. PMID- 8220897 TI - Lack of effect of L-687,414 ((+)-cis-4-methyl-HA-966), an NMDA receptor antagonist acting at the glycine site, on cerebral glucose metabolism and cortical neuronal morphology. AB - 1. N-methyl-D-aspartate (NMDA) receptor ion channel antagonists have been reported to cause pronounced increases in cerebral glucose metabolism (CMRglc) and transient reversible vacuolation within pyramidal cortical neurones. The present studies examined in rats the effects of the NMDA receptor antagonist, L 687,414 (R-(+)-cis-4-methyl-3-amino-l-hydroxypyrolid-2-one (+)-cis-4-methyl-HA 966) on regional CMRglc and cortical neuronal morphology. 2. L-687,414 was given as a steady state intravenous infusion for 4 h in a neuroprotective dose regime of 17.5 mg free base kg-1 bolus followed by 225 micrograms kg-1 min-1 (n = 8) or at the higher dose rate of 35 mg kg-1 bolus followed by 440 micrograms kg-1 min-1 (n = 10). Data were compared to a parallel series of experiments in rats given the NMDA receptor ion channel antagonist, dizocilpine for 4 h in the optimum intravenous neuroprotective dose-regime of 0.12 mg kg-1 bolus followed by 1.8 micrograms kg-1 min-1 (n = 8) or at the higher dose rate of 0.4 mg kg-1 bolus followed by 6 micrograms kg-1 min-1 (n = 4; morphology only studied). A saline infused group of rats (n = 8) were used as controls. 3. CMRglc was studied by use of [14C]-2-deoxyglucose and autoradiography (n = 4 each group) whilst plasma drug levels were in a steady state during the final 45 min of the 4 h drug infusion. Effects on cortical neuronal morphology were assessed at the end of the 4 h infusion period using light microscopic techniques (n = 4-6 each group).4. The results showed a selective activation of limbic CMRglc by dizocilpine at optimal neuroprotective dose levels and showed that this dose was at the threshold for the neuronal vacuolation response as I of 4 rats showed morphological changes in the pyramidal neurones in the posterior cingulate and retrosplenial cortices. At the higher dose rate of dizocilpine, all 4 animals showed extensive morphological changes in these cortical neurones. In contrast, L-687,414 did not increase limbic CMRglc, nor evoke vacuolation when given in the neuroprotective dose regime or at the higher dosage rate.5. The findings of the present study suggest that neuroprotection mediated through the NMDA receptor complex can be achieved without changes in CMRglc or cortical neuronal morphology by antagonism at the glycine modulatory site. PMID- 8220898 TI - Comparison of contractile responses to 5-hydroxytryptamine and sumatriptan in human isolated coronary artery: synergy with the thromboxane A2-receptor agonist, U46619. AB - 1. The interaction between the thromboxane A2 receptor agonist, U46619 and two 5 hydroxytryptamine (5-HT) receptor agonists, the non-selective, naturally occurring agonist, 5-HT and the selective 5-HT1-like agonist, sumatriptan were studied in human epicardial coronary arteries in vitro. 2. Coronary artery rings (2-4 mm in diameter) were prepared from epicardial arteries from explant hearts of patients undergoing heart transplant (cardiomyopathy, n = 13; ischaemic heart disease, n = 10) and unused donor hearts (n = 5). Each ring of artery was set at optimal resting conditions to record changes in isometric force. 3. The majority of artery rings developed phasic, rhythmic contractions either spontaneously or in response to all vasoconstrictor agonists tested. Both the spontaneous and agonist-induced phasic contractions were abolished by nifedipine (0.1 microM). 4. Concentration-contraction curves to 5-HT-receptor agonists and noradrenaline (NA), were first constructed in artery rings that did not develop phasic activity. 5-HT and ergometrine were the most potent agonists with EC50 values of 6.8 +/- 0.2 and 7.7 +/- 0.2 (-log M) respectively. Potencies (EC50's) to sumatriptan, methysergide and noradrenaline could not be determined due to their poor ability to contract the coronary artery. Maximum contractions (Emax; normalized as a percentage of the contraction to a maximum-depolarizing concentration of K+ in physiological salt solution (KPSS)) for 5-HT, ergometrine, sumatriptan, methysergide and noradrenaline were 40 +/- 10, 9 +/- 3, < 5, < 5 and < 5% respectively. 5. In arteries without phasic activity, U46619 (1 nM) caused an increase in force of 3.8 +/- 1% KPSS. With U46619 present, the Emax values for 5-HT, ergometrine, sumatriptan and methysergide were all markedly increased. For 5-HT and sumatriptan, E., values were 92+/- 4% and 49 +/- 14% KPSSrespectively. The presence of U46619 did not significantly change the sensitivity (EC50) to 5 HT.6. In a separate series of arteries, nifedipine (0.1 microM) was used to block phasic, contractile activity. The synergy observed between U46619 and 5-HT or sumatriptan still occurred although the Emax values for each agonist were depressed but the EC50 values were again unaffected.7. In conclusion, these in vitro studies indicate that the normally poor contractions to sumatriptan, inhuman coronary arteries are significantly enhanced when active force is induced with a thromboxane A2-receptor agonist, U46619. The enhanced response is not specific for either sumatriptan or 5-HT,-like receptors since contractions to 5 HT, ergometrine and methysergide were also potentiated by U46619. PMID- 8220899 TI - Involvement of cholecystokinin receptor types in pathways controlling oxytocin secretion. AB - 1. Intravenous administration of cholecystokinin (CCK) results in a transient activation of oxytocin neurones in the rat, and hence to oxytocin secretion: this activation is followed by expression of c-fos mRNA and of Fos-like immunoreactivity (Fos-LI) in magnocellular oxytocin neurones. Fos-like immunoreactivity is also induced in the regions of the brainstem that are thought to relay information from the periphery to the hypothalamus. 2. Administration of the selective CCKA receptor antagonist MK-329, but not the CCKB receptor antagonist L-365,260, prior to CCK injection, prevented oxytocin release as measured by radioimmunoassay and oxytocin neuronal activation as measured by electrophysiology and by the lack of induction of c-fos mRNA. 3. MK-329 abolished the release of adrenocorticotrophic hormone (ACTH) following injection of CCK. 4. MK-329 prevented the expression of Fos-LI in the hypothalamic magnocellular nuclei and in the area postrema and dorsal vagal complex of the brainstem. 5. L 365,260 had no effect on the expression of Fos-LI in the brainstem, but attenuated that seen in the hypothalamic magnocellular nuclei. 6. We conclude that CCK acts on CCKA receptors, either in the area postrema or on peripheral endings of the vagus nerve, to cause the release of hypothalamic oxytocin and ACTH. Information may be carried to the hypothalamus in part by CCK acting at CCKB receptors. PMID- 8220900 TI - Effect of trimebutine on voltage-activated calcium current in rabbit ileal smooth muscle cells. AB - 1. The effect of trimebutine on the voltage-dependent inward Ca2+ current was investigated by the whole-cell voltage-clamp technique in single smooth muscle cells from rabbit ileum. 2. Trimebutine (3-100 microM) reduced the Ca2+ current in a concentration-dependent manner. The inhibitory effect on the Ca2+ current was also dependent on the holding potential. The Ca2+ current after a low holding potential was inhibited to a greater extent than that after a high membrane potential: the IC50 values were 7 microM and 36 microM at holding potentials of 40 mV and -60 mV, respectively. The Ca2+ current elicited from a holding potential of -80 mV could not be reduced by as much as 50% of the control by trimebutine at concentrations as high as 100 microM. 3. Trimebutine (30 microM) shifted the voltage-dependent inactivation curve for the Ca2+ current by 18 mV in the negative direction. The affinity of the drug for Ca2+ channels was calculated to be 36 times higher in the inactivated state than in the closed-available state. 4. Blockade of the Ca2+ current by trimebutine, unlike verapamil, was not use-dependent. 5. The results suggest that trimebutine inhibits the voltage dependent inward Ca2+ current through a preferential binding to Ca2+ channels in the inactivated state in the smooth muscle cell from rabbit ileum. The inhibitory effect of trimebutine on gastrointestinal motility is discussed in the light of the present findings. PMID- 8220901 TI - Nitric oxide-dependent and -independent hyperaemia due to calcitonin gene-related peptide in the rat stomach. AB - 1. Calcitonin gene-related peptide (CGRP) potently enhances mucosal blood flow in the rat stomach. The aim of this study was to examine whether CGRP also dilates extramural arteries supplying the stomach and whether the vasodilator action of CGRP involves nitric oxide (NO). 2. Rat CGRP-alpha (0.03-1 nmol kg-1, i.v.) produced a dose-dependent increase in blood flow through the left gastric artery (LGA) as determined by an ultrasonic transit time technique in urethane anaesthetized rats. Blockade of NO synthesis by NG-nitro-L-arginine methyl ester (L-NAME, 20 and 60 mumol kg-1, i.v.) significantly reduced basal blood flow (BF) in the LGA and attenuated the hyperaemic activity of CGRP by a factor of 2.8-4. D NAME tended to enhance basal BF in the LGA but had no influence on the dilator activity of CGRP. The ability of vasoactive intestinal polypeptide to increase left gastric arterial blood flow remained unaltered by L-NAME. 3. L-NAME (20 and 60 mumol kg-1, i.v.) evoked a prompt and sustained rise of mean arterial blood pressure (MAP) and caused a slight decrease in the hypotensive activity of CGRP. In contrast, D-NAME induced a delayed and moderate increase in MAP and did not influence the hypotensive activity of CGRP. 4. Rat CGRP-alpha dilated the isolated perfused bed of the rat LGA precontracted with methoxamine and was 3 times more potent in this respect than rat CGRP-beta. The dilator action of rat CGRP-alpha in this preparation was not affected by L-NAME or D-NAME (40 microM). 5. L-NAME (60 micromol kg-1, i.v.) reduced gastric mucosal blood flow as assessed by laser Doppler flowmetry and diminished the hyperaemic activity of rat CGRP alpha in the gastric mucosa by a factor of 4.5, whereas D-NAME was without effect.6. These data show that CGRP is a potent dilator of mucosal and extramural resistance vessels in the rat stomach. Its dilator action involves both NO dependent and NO-independent mechanisms. PMID- 8220902 TI - Bradykinin-induced release of PGI2 from aortic endothelial cell lines: responses mediated selectively by Ca2+ ions or a staurosporine-sensitive kinase. AB - 1. Bradykinin (100 nM) triggers release of nitric oxide and prostacyclin from both AG07680A and AG04762 bovine cultured aortic endothelial cells. The exposure of these cells to bradykinin is in each case associated with a striking rise in intracellular calcium ion concentration. 2. Exposure of AG07680A cells to 250 nM ionomycin was followed also by a significant release of prostacyclin, whereas 250 nM ionomycin had no capacity to stimulate release of prostacyclin from AG04762 cells. 3. There was a similar concentration-dependent increase in intracellular calcium ion concentration on exposure of AG07680A and AG04762 cells to ionomycin. 4. Exposure of AG04762 cells for 10 min to staurosporine produced a concentration dependent inhibition (IC50 = 107 +/- 14 nM) in bradykinin-stimulated prostacyclin release. There was no similar inhibitory effect of staurosporine in AG07680A cells. 5. Bradykinin (10 nM) triggered release of nitric oxide from both AG07680A and AG04762 cells, and the effect was not inhibited by 500 nM staurosporine. There was a similar ionomycin-dependent release of nitric oxide from both cell types. 6. These results identify a common pathway for bradykinin-dependent nitric oxide release from both AG07680A and AG04762 cells, involving increases in intracellular calcium ion concentration. In contrast, the bradykinin-dependent release of prostacyclin may involve one of two pathways (involving an increase in intracellular calcium or activation of a staurosporine-sensitive kinase), and the two pathways are selectively exploited in AG07680A and AG04762 cells, respectively. PMID- 8220903 TI - Specific inhibition of leukotriene B4 (LTB4)-induced neutrophil emigration by 20 hydroxy LTB4: implications for the regulation of inflammatory responses. AB - 1. The interaction between leukotriene B4 (LTB4) and its metabolite, 20-hydroxy LTB4 in the control of neutrophil emigration was examined in guinea-pig skin. 2. Leukotriene B4 (10-300 ng) elicited a dose-dependent increase in neutrophil infiltration (as measured by myeloperoxidase activity) 4 h after injection into guinea-pig skin. In contrast, 20-hydroxy LTB4 (30-1000 ng) displayed only weak inflammatory activity in this assay. 3. Although 20-hydroxy LTB4 had low agonist activity, this metabolite caused a potent dose-dependent inhibition of responses to LTB4 (100 ng), when administered systemically (ED50 = 1.3 micrograms kg-1, s.c.) without significantly affecting neutrophil infiltration in response to C5a (2 micrograms). Systemic administration of 20-carboxy LTB4 (10 micrograms) did not affect neutrophil accumulation in response to LTB4 or C5a. In addition, neither 15(S)-hydroxy 5(S)-HPETE(10 micrograms) nor lipoxin A4 (10 micrograms) inhibited responses to LTB4. 4. Addition of 20-hydroxy LTB4 (10(-11)-10(-8) M) to human blood prior to isolation of the neutrophils led to concentration-dependent decrease in the number of LTB4 receptors and decreased chemotactic responsiveness to LTB4 without affecting responses to C5a. Incubation of blood with 20-carboxy LTB4 (10(-8) M) did not reduce LTB4 receptor number of chemotactic responsiveness to LTB4. 5. These data indicate that although 20-hydroxy LTB4 is a weak agonist at LTB4 receptors, it can desensitize neutrophils to the effects of LTB4 via down regulation of the high affinity receptor and thus provides evidence for a mechanism whereby inflammatory responses may be regulated. PMID- 8220904 TI - Cerebral blood flow and cerebrovascular reactivity after inhibition of nitric oxide synthesis in conscious goats. AB - 1. The role of nitric oxide in the cerebral circulation under basal conditions and after vasodilator stimulation was studied in instrumented, conscious goats, by examining the action of inhibiting endogenous nitric oxide production with NG nitro-L-arginine methyl ester (L-NAME). 2. In 6 unanaesthetized goats, blood flow to one brain hemisphere (electromagnetically measured), systemic arterial blood pressure and heart rate were continuously recorded. L-NAME (35 mg kg-1 by i.v. bolus) decreased resting cerebral blood flow by 43 +/- 3%, increased mean arterial pressure by 21 +/- 2%, and decreased heart rate by 41 +/- 2%; cerebrovascular resistance increased by 114 +/- 13% (P < 0.01); the immediate addition of i.v. infusion of L-NAME (0.15-0.20 mg kg-1 during 60-80 min) did not significantly modify these effects. Cerebral blood flow recovered at 72 h, arterial pressure and cerebrovascular resistance at 48 h, and heart rate at 6 days after L-NAME treatment. 3. A second treatment with L-NAME scheduled as above reproduced the immediate haemodynamic effects of the first treatment, which (except bradycardia) reversed with L-arginine (200-300 mg kg-1 by i.v. bolus). 4. Acetylcholine (0.01-0.3 micrograms), sodium nitroprusside (3-100 micrograms) and diazoxide (0.3-9 mg), injected into the cerebral circulation of 5 conscious goats, produced dose-dependent increases in cerebral blood flow, and decreases in cerebrovascular resistance; sodium nitroprusside (30 and 100 micrograms) also caused hypotension and tachycardia. 5. The reduction in cerebrovascular resistance from resting levels (in absolute values) to lower doses,but not to the highest dose, of acetylcholine was diminished, to sodium nitroprusside was increased, and to diazoxide was unaffected after L-NAME, compared to control conditions. The effects on cerebrovascular resistance to acetycholine normalized within 24 h and to sodium nitroprusside within 48 h after L-NAME treatment.6. This study provides information about the evolution of the changes in cerebral blood flow and cerebrovascular reactivity after inhibition of endogenous nitric oxide in conscious animals. The results suggest: (a) endogenous nitric oxide is involved in regulation of the cerebral circulation by producing a resting vasodilator tone, (b) the cerebral vasodilatation to acetylcholine is mediated, at least in part, by nitric oxide release, and (c) inhibition of nitric oxide production induces supersensitivity of cerebral vasculature to nitrovasodilators. PMID- 8220905 TI - Endothelin-1 (ET-1)-induced contraction in rat isolated trachea: involvement of ETA and ETB receptors and multiple signal transduction systems. AB - 1. Quantitative autoradiographic, biochemical and functional studies were performed to investigate the endothelin receptor subtypes and signal transduction systems that mediate endothelin-1 (ET-1)-induced contraction in rat isolated tracheal smooth muscle. 2. Specific binding of 0.5 nM [125I]-ET-1 to tracheal smooth muscle was inhibited by at least 40% in the presence of either the ETA receptor selective ligand BQ-123 (1 microM) or the ETB receptor-selective ligand sarafotoxin S6c (30 nM), indicating the presence of both ETA and ETB receptors in this tissue. 3. ET-1 and sarafotoxin S6c were both potent spasmogens of rat isolated tracheal smooth muscle preparations. Sarafotoxin S6c-induced contractions were unaffected in the presence of the ETA receptor antagonist BQ 123 (10 microM), but were markedly attenuated in tissue previously exposed to 100 nM sarafotoxin S6c to induce ETB receptor desensitization. ET-1-induced contractions were, at most, only partially attenuated either by blocking the ETA receptor-effector system (with 10 microM BQ-123) or by desensitizing the ETB receptor-effector system with sarafotoxin S6c. However, ET-1-induced contractions were markedly attenuated by blocking both receptor-effector systems simultaneously. These findings suggest that ET-1 could induce contraction by stimulating either ETA or ETB receptors. 4. ET-1 (10 microM) induced a 7 fold increase in intracellular [3H]-inositol phosphate accumulation over basal levels in rat isolated tracheal smooth muscle. In contrast, sarafotoxin S6c (2.5 microM) increased intracellular [3H]-inositol phosphate accumulation by only 2 fold. ET-1 induced accumulation of [3H]-inositol phosphates was abolished by 10 microM BQ 123. 5. In Ca2+-free Krebs bicarbonate solution, 100 nM ET-1 induced a significantly larger contraction than that induced by 100 nM sarafotoxin S6c (46.6 +/- 5.6% C,., versus 8.8 +/- 2.8% Cmax, n = 5-7). This presumed intracellular Ca2+-dependent phase of contraction induced by ET-1 was significantly inhibited by 10 microM BQ-123 (7.5 +/- 1.0% C.). Subsequent addition of 2.5 mM Ca2+ induced a second phase of contraction. The extracellular Ca2+-dependent phase of contraction induced by ET-1 was similar inmagnitude to that induced by sarafotoxin S6c (63.6 +/- 4.5% C.. versus 58.0 +/- 3.7% C.) and was not inhibited by BQ-123. Sarafotoxin S6c-induced contractions were not inhibited by the L-type Ca2+-channel antagonists, nicardipine or verapamil.6. In summary, ETA and ETB receptors coexist in rat isolated tracheal smooth muscle and stimulation of both receptor subtypes contributes to ET-l-induced contraction in this tissue. However, stimulation of these receptor subtypes appears to induce contraction by activating different second messenger pathways; ETA receptor stimulation induces phosphoinositide turnover and subsequent release of intracellular Ca2+ whereas stimulation of ETB receptors facilitates the influx of extracellular Ca2+. PMID- 8220906 TI - Suppression of inflammatory responses to 12-O-tetradecanoyl-phorbol-13-acetate and carrageenin by YM-26734, a selective inhibitor of extracellular group II phospholipase A2. AB - 1. YM-26734 [4-(3,5-didodecanoyl-2,4,6-trihydroxyphenyl)-7-hydroxy-2-(4-hydroxyph eny l) chroman] dose-dependently inhibited the activities of extracellular phospholipase A2 (PLA2): rabbit platelet-derived group II and porcine pancreas derived group I PLA2, with IC50 values of 0.085 (0.056-0.129, n = 5) and 6.8 (5.0 9.6, n = 5) microM, respectively. 2. In contrast, YM-26734 did not reduce the activity of intracellular PLA2 prepared from mouse macrophages, which preferentially hydrolyzed arachidonoyl phospholipids at concentrations up to 50 microM. YM-26734 also showed no effect against either sheep seminal vesicle cyclo oxygenase or rat leukocyte 5-lipoxygenase. 3. Linewater-Burk analysis showed that YM-26567-1 behaved as a competitive inhibitor of group II PLA2 derived from rabbit platelets, with a Ki value of 48 nM. 4. In mice, YM-26734 inhibited 12-O tetradecanoylphorbol-13-acetate (TPA, 1 microgram/ear)-induced ear oedema in a dose-dependent manner, with ED50 values of 45 (30-67) micrograms/ear (n = 5) and 11 (4-32) mg kg-1, i.v. (n = 5), but did not decrease arachidonic acid (4 mg/ear) induced ear oedema at 1 mg/ear and 30 mg kg-1, i.v. 5. In rats, the accumulation of exudate fluids and leukocytes in the pleural cavity in response to carrageenin injection (2 mg) was significantly less in a group treated with YM-26734 (20 mg kg-1, i.v.) than in the control group (0.43 +/- 0.02 vs 0.59 +/- 0.03 g per cavity and 3.8 +/- 0.2 vs 4.9 +/- 0.3 x 10(7) cells per cavity, respectively; n = 5). 6. These results suggest that YM-26734 is a potent and competitive inhibitor of extracellular PLA2 with selectivity for group II PLA2, and that the inhibition of group II enzymes activity may cause the suppression of inflammatory responses to TPA and carrageenin. PMID- 8220907 TI - Vascular actions of purines in the foetal circulation of the human placenta. AB - 1. The vasoactive effects of adenosine triphosphate (ATP), adenosine and other purines in the foetal circulation of the human placenta were examined. Single lobules of the placenta were bilaterally perfused in vitro with Krebs buffer (maternal and foetal sides 5 ml min-1 each, 95% O2:5% CO2, 37 degrees C). Changes in foetal vascular tone were assessed by recording perfusion pressure during constant infusion of each purine. To allow recording of the vasodilator effects, submaximal vasoconstriction was induced by concomitant infusion of prostaglandin F2 alpha (0.7-2.0 mumol l-1). 2. ATP (1.0-100 mumol l-1) usually caused concentration-dependent reductions in perfusion pressure. However, biphasic with initial transient increases, or only increases in pressure were sometimes observed. Falls in pressure caused by ATP were significantly reduced by addition to the perfusate of NG-nitro-L-arginine (L-NOARG) (100 mumol l-1) but not NG nitro-D-arginine (D-NOARG) (100 mumol l-1). They were not influenced by addition of indomethacin (10 mumol l-1) or L-arginine (100 mumol l-1). 3. Adenosine (0.01 1.0 mmol l-1) consistently caused concentration-dependent reductions in perfusion pressure, this effect not being influenced by indomethacin. L-NOARG, but not D NOARG, reduced the potency of adenosine approximately three fold. L-Arginine, but not D-arginine enhanced its potency by a similar amount. 4. 2-Methylthio-ATP, a selective P2 gamma agonist was approximately 50 times more potent than ATP as a vasodilator agent, always causing decreases in perfusion pressure. 5. Beta-gamma Methylene ATP, a selective P20 agonist, was approximately 100 times more potent than ATP as a vasoconstrictor, but only caused transient increases in perfusion pressure.6. The rank order of vasodilator potencies of a selection of adenosine receptor agonists was, 2-chloroadenosine>>5-(N-cyclopropyl)-carboxamidoadenosine, >5-N-ethylcarboxamidoadenosine, >2-chloro-N6-cyclopentyladenosine, >CGS-21680 > N6-cyclohexyladenosine = adenosine. Vasodilatation due to adenosine was inhibited by the PI-A2 receptor antagonist 3,7-dimethyl-l-propargylxanthine(DMPX).7. These results suggest that ATP may cause an endothelium-dependent vasodilatation in the foetal vessels of the human placenta via activation of a P2y receptor linked to the formation of nitric oxide(NO). Vasodilatation caused by ATP may mask an accompanying vasoconstrictor effect mediated, via a P2X receptor, in the villous vascular smooth muscle. Adenosine acting on P1-A2 receptors, which are also present in the foetal vasculature, may require synergistic interaction with NO to achieve a maximal vasodilator response. PMID- 8220908 TI - The effect of ions and second messengers on long-term potentiation of chemical transmission in avian ciliary ganglia. AB - 1. The effects of tetanic stimulation of the oculomotor nerve on transmission through the avian ciliary ganglion have been determined by use of the amplitude of the compound action potential recorded in the ciliary nerve, in the presence of hexamethonium (300 microM), as a measure of synaptic efficacy. 2. Tetanic stimulation for 20 s at 30 Hz potentiated the chemical phase of the compound action potential by at least 100% of its control level. This potentiation, reflecting an increase in synaptic efficacy, decayed over two distinct time courses: firstly, a rapid decay with a time constant in the order of minutes, and secondly, a slower decay, representing a smaller potentiation, with a time constant in the order of an hour. The large increase in synaptic efficacy is attributed to post-tetanic potentiation (PTP) whereas the smaller but longer lasting increase is attributed to long-term potentiation (LTP). 3. Higher frequencies of tetanic stimulation gave increased PTP and LTP. 4. In order to test whether the influx of calcium ions into the nerve terminal during the tetanus is likely to be involved in potentiation, facilitation was measured during PTP and LTP. Facilitation was reduced to approximately zero during PTP but recovered to normal values about 15 min into LTP. A requirement for the induction of LTP was shown to be the presence of calcium in the bathing solution. However, blocking synaptic transmission with a high concentration of hexamethonium (3 mM) during the tetanic stimulation did not block the induction of LTP. 5. Application of the muscarinic inhibitor, atropine (2 microM), did not affect the magnitude of PTP or LTP. 5. Application of the muscarinic inhibitor, atropine (2 tM), did not affect the magnitude of PTP or LTP.6. The activator of protein kinase C, phorbol 12,13-dibutyrate (2 microM) potentiated synaptic transmission and reduced the potentiation due to PTP although it did not affect that due to LTP, but the inhibitor of this kinase, staurosporine (0.5 microM), partially blocked the appearance of LTP without affecting PTP after the tetanus.7. An inhibitor of calmodulin, W-7 (5 microM), reversibly blocked the appearance of LTP significantly after a tetanus although the size of PTP was not affected.8. The results presented here suggest that the initiation of LTP in the ciliary ganglion is due to an influx of calcium ions into the calyciform nerve terminal during the tetanus and that the mechanism for LTP involves a calcium-calmodulin-dependent process. PMID- 8220909 TI - Mediation of the neuroprotective action of R-phenylisopropyl-adenosine through a centrally located adenosine A1 receptor. AB - 1. Systemic injections of kainic acid, 10 mg kg-1, into adult rats resulted in lesions in the hippocampus, as assessed by peripheral benzodiazepine ligand binding. Co-administration of clonazepam at 1 mg kg-1 or 0.2 mg kg-1 prevented major seizures associated with kainate injections, but did not alter significantly the production of hippocampal damage. 2. The co-administration of the adenosine A1 agonist R-phenylisopropyladenosine (R-PIA, 25 micrograms kg-1, i.p.) abolished the lesions induced by kainic acid. 3. The presence of the selective A1 antagonist, 8-cyclopentyl-1,3-dipropylxanthine (250 or 50 micrograms kg-1, i.p.) abolished the R-PIA neuroprotective action. 4. The A1/A2 antagonist, 8-(p-sulphophenyl)theophylline (20 mg kg-1, i.p.) which cannot cross the blood brain barrier, did not alter significantly the neuroprotective action of R-PIA, indicating that the neuroprotective action of the purine may be predominantly central. 5. The time course of the neuroprotection was also examined. R-PIA was effective when administered 2 h before or after kainate administration. 6. The results emphasise the potential utility of systemically active adenosine A1 receptor ligands in reducing CNS gliosis induced by the activation of excitatory amino acid receptors. PMID- 8220910 TI - Differential effects of B2 receptor antagonists upon bradykinin-stimulated phospholipase C and D in guinea-pig cultured tracheal smooth muscle. AB - 1. Guinea-pig tracheal smooth muscle cells were isolated and maintained in culture for 14-21 days prior to the study of the effect of a selective bradykinin B1 agonist and B2 antagonists upon bradykinin-stimulated phospholipase C and D activities. 2. Bradykinin-stimulated phospholipase C activity was determined by mass measurement of inositol (1,4,5)trisphosphate (Ins(1,4,5)P3) in unlabelled cells, whereas phospholipase D activity was assayed by the accumulation of [3H] phosphatidylbutanol ([3H]-PtdBut) in [3H]-palmitate-labelled cells, which were stimulated in the presence of butan-1-o1 (0.3%, v/v). 3. Bradykinin elicited the rapid and transient formation of Ins(1,4,5)P3, in a concentration-dependent manner (log EC50 = -7.55 +/- 0.1 M, N = 3). Bradykinin also rapidly activated the concentration-dependent (log EC50 = -8.3 +/- 0.4 M, n = 3) phospholipase D catalysed accumulation of [3H]-PtdBut; the accumulation of [3H]-PtdBut was sustained. These effects were not inhibited by pretreatment of the cells with indomethacin (1 microM). 4. The bradykinin B1 agonist, desArg9-bradykinin (1 microM) was without effect upon phospholipase C or phospholipase D activity. Bradykinin-stimulated (10 nM, EC40) Ins(1,4,5)P3 formation was inhibited by B2 receptor antagonists, D-Arg-[Hyp3,D-Phe7]-bradykinin (NPC 567) and D-Arg [Hyp3,Thi5,8,D-Phe7]-bradykinin (NPC 349), with log IC50 values of -6.3 +/- 0.5 M and -6.3 +/- 0.4 M, respectively. However, bradykinin-stimulated (10 nM, EC100) [3H]-PtdBut accumulation was poorly inhibited and with low potency by each B2 receptor antagonist and bradykinin-stimulated phospholipase D activity persisted at concentrations of antagonist that completely blocked bradykinin-stimulated Ins(1,4,5)P3 formation (30 microM). 5. These observations suggest that the activation of phospholipase C by bradykinin may be mediated through a bradykinin B2 receptor population, whereas bradykinin-stimulated phospholipase D may be activated via a distinct population of bradykinin receptors that do not appear to be either B1 or B2 receptor types, based upon pharmacological specificity. The mechanism of the activation of phospholipase D by bradykinin and the role of the putative B3 bradykinin receptor are discussed. PMID- 8220912 TI - Antitussive effects of GABAB agonists in the cat and guinea-pig. AB - 1. GABAB agonists inhibit neuronal processes which are important in the pathogenesis of airway disease, such as bronchospasm. Cough is a prominent symptom of pulmonary disease, but the effects of GABAB agonists on this airway reflex are unknown. Experiments were conducted to determine the antitussive effect of GABAB receptor agonists in comparison to the known antitussive agents, codeine and dextromethorphan. 2. Unanaesthetized guinea-pigs were exposed to aerosols of 0.3 mM capsaicin to elicit coughing, which was detected with a microphone and counted. Cough also was produced in anaesthetized cats by mechanical stimulation of the intrathoracic trachea and was recorded from electromyograms of respiratory muscle activity. 3. In guinea-pigs, the GABAB agonists baclofen and 3-aminopropyl-phosphinic acid (3-APPi) produced dose dependent inhibition of capsaicin-induced cough when administered by subcutaneous or inhaled routes. The potencies of baclofen and 3-APPi compared favourably with codeine and dextromethorphan. 4. The GABAB antagonist, CGP 35348 (0.3- 30 mg kg 1, s.c.) inhibited the antitussive effect of baclofen (3.0 mg kg-1, s.c.). However, CGP 35348 (10 mg kg-1, s.c.) had no effect on the antitussive activity of codeine (30 mg kg-1, s.c.). The antitussive effect of baclofen was not influenced by the GABAA antagonist, bicuculline (3 mg kg-1, s.c.) or naloxone (0.3 mg kg-1, s.c.). 5. In the cat, baclofen (0.3-3.0 mg kg-1, i.v.) decreased mechanically-induced cough in a dose-dependent manner. In this model, baclofen (ED50 = 0.63 mg kg-1) was less potent than either codeine or dextromethorphan. The antitussive effect of baclofen in the cat was antagonized by the GABAB antagonists, CGP 35348 (10 mg kg-1, i.v.) and 3-aminopropylphosphonic acid (3 mg kg-1, i.v.).6. We show that baclofen and 3-APPi have antitussive effects in the guinea-pig and cat and these effects are mediated by GABAB receptors. PMID- 8220911 TI - Human liver microsomal metabolism of the enantiomers of warfarin and acenocoumarol: P450 isozyme diversity determines the differences in their pharmacokinetics. AB - 1. To explain the large differences in (the stereoselectivity of) the clearances of the enantiomers of warfarin and acenocoumarol (4'-nitrowarfarin) their human liver microsomal metabolism has been studied and enzyme kinetic parameters determined. The effects of cimetidine, propafenone, sulphaphenazole, and omeprazole on their metabolism has been investigated. 2. The 4-hydroxycoumarins follow similar metabolic routes and are mainly hydroxylated at the 6- and 7 position (accounting for 63 to 99% of the metabolic clearances). 3. Due to the lower Km values of R- and S-acenocoumarol and higher Vmax values of S acenocoumarol, the overall metabolic clearances of R/S acenocoumarol exceed those of R/S warfarin 6 and 66 times respectively. 4. The metabolism of both compounds is stereoselective for the S-enantiomers, which is 10 times more pronounced in the case of acenocoumarol. 5. Except for the 7-hydroxylation of the R-enantiomers (r = 0.90; P < 0.025), the 6- and 7-hydroxylation rates of R/S warfarin do not correlate with those of R/S acenocoumarol. 6. Sulphaphenazole competitively inhibits the 7- and in some samples partly (up to 50%) the 6-hydroxylation of S warfarin as well as the 7-hydroxylation of R- and S-acenocoumarol and the 6 hydroxylation of S-acenocoumarol (Kis ranging from 0.5-1.3 microM). 7. Omeprazole partly (40-80%) inhibits the 6- and 7-hydroxylation of R-warfarin (Ki = 99 and 117 microM) and of R- (Ki = 219 and 7.2 microM) and S-acenocoumarol (Ki = 6.1 and 7.7 microM) but not S-warfarin in a competitive manner. 8. Differences in the partial (up to 40%) inhibition of the metabolism of the enantiomers of the 4 hydroxycoumarins were also observed for the relatively weak inhibitors, propafenone and cimetidine.9. The results suggest that the coumarin ring hydroxylations of both compounds are catalysed by different combinations of P450 isozymes. The 7-hydroxylation of R/S acenocoumarol and the 6-hydroxylation of S acenocoumarol are at least partly conducted by (a) P450 isozyme(s) of the 2C subfamily different from P450 2C9 (the main S-warfarin 7- and 6-hydroxylase). PMID- 8220913 TI - The paradoxical vascular interactions between endothelin-1 and calcitonin gene related peptide in the rat gastric mucosal microcirculation. AB - 1. The interactions between local intra-arterial infusion of endothelin-1 (ET-1) and rat alpha-calcitonin gene-related peptide (alpha-CGRP) on gastric mucosal damage and blood flow have been investigated in the pentobarbitone-anaesthetized rat. 2. Close-arterial infusion of ET-1 (2-200 pmol kg-1 min-1) induced a significant and dose-dependent increase in gastric mucosal haemorrhagic injury. 3. Close-arterial infusion of the higher doses of ET-1 (100 and 200 pmol kg-1 min 1) resulted in a biphasic effect on mucosal blood flow, as determined by laser Doppler flowmetry (LDF). This consisted of an initial transient increase followed by a pronounced and sustained fall in LDF. 4. Local microvascular constriction may thus contribute to the mechanisms underlying the gastric injury induced by these higher doses of ET-1. 5. However, close-arterial infusion of lower doses of ET-1 (2-50 pmol kg-1 min-1), that also provoked substantial mucosal damage, induced only a sustained and significant mucosal hyperaemia, which may be secondary to microvascular injury. 6. Concurrent dose-arterial administration of rat alpha-CGRP (50 pmol kg-1 min-1) significantly inhibited the extent of gastric mucosal injury induced by ET-1 (5 pmol kg-1 min-1). 7. Furthermore, concurrent close-arterial infusion of this dose of alpha-CGRP, which itself increased mucosal LDF, significantly inhibited the hyperaemic response induced by close arterial infusion of ET-1 (5 pmol kg-1 min-1). 8. These results indicate a damaging action on the gastric mucosa by low doses of ET-1 which is independent of local vasoconstriction, that may involve a direct injury of the microvascular endothelium. The protective action of alpha-CGRP thus seems unlikely to be due to a local vasodilator effect but may reflect protective actions on the microvascular endothelium PMID- 8220914 TI - Glycine stimulates striatal dopamine release in conscious rats. AB - 1. Glycine is an inhibitory neurotransmitter in the spinal cord and brainstem. The mechanism of this inhibition is via binding of glycine to specific receptors, increasing transmembrane Cl- conductance and hyperpolarizing neurones. Strychnine selectively antagonizes these effects. The role of glycinergic neurones in supraspinal regions is poorly understood. 2. Effects of glycine on release of catecholamines in the striatum were examined by microdialysis in freely-moving rats. Transcription of the genes encoding strychnine-sensitive glycine receptors was assessed in the striatum and substantia nigra, by use of reverse transcription followed by the polymerase chain reaction. 3. Glycine administered via the microdialysis probe dose-dependently increased concentrations of dopamine and its metabolites, dihydroxyphenylacetic acid and homovanillic acid, in the perfusate, indicating increased local release and metabolism of dopamine. Strychnine markedly attenuated these responses. Whereas striatal tissue did not contain mRNA for either the adult or neonatal form of strychnine-sensitive glycine receptor, nigral tissue contained a message for the adult form. 4. The results suggest that dopaminergic cells in the substantia nigra synthesize strychnine-sensitive glycine receptors and transport the receptors to terminals in the striatum. Occupation of the glycine receptors then exerts a net stimulatory effect on striatal dopamine release in vivo. PMID- 8220915 TI - Desensitization of the P2-purinoceptors on the rat colon muscularis mucosae. AB - 1. Adenosine 5'-triphosphate (ATP) and adenosine have been shown to contract the rat colon muscularis mucosae, and the receptors at which they act have been classified as P2Y and A1 respectively. Uridine 5'-triphosphate (UTP) also contracts this tissue, and desensitization was used to investigate the receptors by which it acts, in the light of recent suggestions that specific pyrimidinoceptors may exist for UTP, or that nucleotide receptors may exist which are responsive to both ATP and UTP but not to some ATP analogues such as 2 methylthioadenosine 5'-triphosphate (2-MeSATP). 2. ATP, UTP and adenosine each contracted the rat colon muscularis mucosae in a concentration-dependent manner over the concentration range 0.3-300 microM, although maximal responses to ATP and UTP were not obtained. ATP was approximately 4 times as potent as UTP and approximately equipotent with adenosine although the maximal response to adenosine appeared to be less than that to ATP or UTP. 3. Desensitization of the tissue with ATP (200 microM) given immediately before each concentration of the agonists reduced subsequent contractions induced by ATP itself and also by UTP, but did not reduce contractions induced by adenosine. Desensitization of the tissues with UTP (200 microM) also reduced contractions induced by ATP and UTP but not by adenosine, whereas desensitization with adenosine (200 microM) reduced contractions induced by adenosine itself but not by ATP or UTP. 4. Desensitization of the tissue with 2-MeSATP (200 microM), which is a more potent agonist than ATP at P2Y-purinoceptors, greatly reduced the responses to ATP and to UTP, but had no effect on responses induced by adenosine. Attempts to desensitize the tissue with adenosine 5'-(alpha,beta methylene)triphosphonate(AMPCPP), which is a more potent agonist than ATP at P2X purinoceptors but is less potent atP2y-purinoceptors, were unsuccessful.5. These results show that cross desensitization to ATP and UTP occurred and was specific for these agonists rather than being due to a general decrease in the ability of the muscle to contract. This implies that ATP and UTP act at the same receptor, which does not support the existence of specificpyrimidinoceptors but which could be taken as evidence for the existence of a nucleotide receptor on this tissue. However, the ability of 2-MeSATP, which is inactive at the proposed nucleotide receptors,also selectively to desensitize this receptor indicates instead that ATP and UTP are both acting at a purinoceptor of the P2Y type in this tissue. PMID- 8220916 TI - Mediation by B1 and B2 receptors of vasodepressor responses to intravenously administered kinins in anaesthetized dogs. AB - 1. Vasodepressor responses to intravenous (i.v.) injection of bradykinin (BK) and des-Arg9-BK, a selective B1 kinin receptor agonist, were characterized following i.v. pretreatment with selective B1 ([Leu8]-des-Arg9-BK) and B2 (Hoe 140) kinin receptor antagonists in anaesthetized dogs. 2. Des-Arg9-BK (0.05-3.3 nmol kg-1) produced dose-dependent decreases in mean arterial blood pressure with a ED50 0.4 nmol kg-1. The vasodepressor effects evoked by des-Arg9-BK (0.6 nmol kg-1) and BK (0.2 nmol kg-1) were greater after i.v. and i.a. injections, respectively. 3. The vasodepressor response to BK (0.6 nmol kg-1) but not to des-Arg9-BK (0.6 nmol kg 1) was significantly (P < 0.001) blocked by pretreatment with the B2 receptor antagonist, Hoe 140. 4. The vasodepressor response to des-Arg9-BK (0.6 nmol kg-1) but not to BK (0.6 nmol kg-1) was significantly (P < 0.001) reduced by pretreatment with the selective B1 receptor antagonist, [Leu8]-des-Arg9-BK. Although both B1 and B2 receptor antagonists caused a transient fall in blood pressure, their inhibitory action was unlikely to be related to a desensitization mechanism. 5. Inhibition of prostaglandin synthesis with indomethacin prevented the vasodepressor response induced by arachidonic acid (1 mg kg-1, i.v.) but not that to BK or des-Arg9-BK (0.6 nmol kg-1). 6. These results suggest, firstly, that the vasodepressor responses to i.v. BK and des-Arg9-BK are mediated by the activation of B2 and B1 receptors, respectively; secondly, that prostaglandins are not involved in the vasodepressor responses to kinins.These findings provide pharmacological evidence for the existence of functionally active B1 receptors in canine cardiovascular homeostasis. PMID- 8220917 TI - Membrane current responses to externally-applied ATP in the longitudinal muscle of the chicken rectum. AB - 1. Membrane current responses to ATP in enzymically-dispersed single smooth muscle cells from the chicken rectum were investigated by the whole-cell voltage clamp technique. 2. In cells dialysed with a KCl-rich solution under voltage clamp at a holding potential of -40 mV, ATP (10 microM) produced an inward current followed by an outward current. When the holding potential was changed to 0 mV and -80 mV, the biphasic current response to ATP was converted to an outward current alone and an inward current alone, respectively. 3. External application of tetraethylammonium (TEA, 5 mM), intracellular dialysis with a CsCl-rich solution, or inclusion of EGTA (10 mM) in the pipette abolished the outward current response to ATP. 4. Neither depletion of Ca2+ store with caffeine (10 mM) nor block of voltage-gated Ca2+ channels with nifedipine (10 microM) affected the biphasic current response to ATP. After removal of the extracellular Ca2+ the outward current response to ATP was abolished. 5. alpha,beta-methylene ATP (100 microM) elicited a current similar to the ATP-induced current. In the presence of alpha,beta-methylene ATP (100 microM), application of ATP (100 microM) was without effect. 6. In CsCl-filled cells, ATP analogues elicited an inward current and the order of potency was ATP not equal to alpha, beta-methylene ATP > ADP >> AMP. 7. Inclusion of GTP gamma S (0.2 mM) or GDP beta S (2 mM) in the pipette did not affect the ATP-induced inward current in CsCl-filled cells. The reversal potential of the ATP-induced inward current was about 0 mV and was completely inhibited after replacement of the cations in the bath solution by Tris. The reversal potential remained almost unchanged after replacement of Na+ in the bath solution with 1 10 mM Ca2+, but shifted in the negative direction after replacement of Na+ or both Na+ and Ca2+ with glucosamine.8. The results suggest that ATP acts on P2 purinoceptors to cause activation of cation channels with selectivity for Ca2+ over Na+. Moreover, it appears that no G-protein-mediated mechanism is involved and increased Ca2+ entry through the cation channels causes activation of Ca2+-activated K+ channels. PMID- 8220919 TI - Local action transcutaneous flurbiprofen in the treatment of soft tissue rheumatism. AB - The objective of the present study was to establish the efficacy and tolerability of local action transcutaneous flurbiprofen (flurbiprofen LAT) in the treatment of soft tissue lesions. A randomized, double-blind, parallel-group placebo controlled study was carried out in two hospital outpatient rheumatology clinics. One hundred and four patients aged 18-75 yr were randomized to receive a non woven polyester-backed patch supporting a formulation containing 40 mg flurbiprofen 12-hourly over 14 days; or a non-medicated (but otherwise identical) control. Statistically significant differences in favour of the active preparation were seen at both days 7 (P = 0.02) and 14 (P = 0.009) for the investigator's overall opinion of severity of the condition, and at day 7 for the investigator's assessment of pain severity (P = 0.04 intention-to-treat; P = 0.052 N.S. eligible data). The need for further treatment in the form of steroid injections after the trial was greater in the controls (29/44, 66%) than in the flurbiprofen LAT group (17/46, 37%) (chi 2 = 7.54 on 1 d.o.f., P = 0.006). Plasma flurbiprofen levels in 11 patients ranged from 13.4 to 338.7 ng/ml (mean 116; median 57.9). Eight out of 53 (15%) patients receiving flurbiprofen LAT reported a total of 10 adverse events, compared with three out of 51 (6%) reporting seven events among controls. Patients found the patch convenient and soothing. We conclude that flurbiprofen LAT is an effective and acceptable treatment for soft tissue lesions, and should be considered as an alternative therapy to local steroid injection. PMID- 8220918 TI - The positive inotropic effect of compound II, a novel analogue of sotalol, on guinea-pig papillary muscles and single ventricular myocytes. AB - 1. Compound II is a novel analogue of sotalol which has been reported to be free of beta-adrenoceptor and L-type calcium channel blocking actions. The effects of compound II on the contraction of guinea-pig papillary muscles (at 2 microM) and single ventricular myocytes (at 100 nM) were investigated. 2. Exposure to compound II caused a significant increase in the contraction of both preparations. 3. Compound II prolonged the action potential of the single myocytes and increased the magnitude of the Ca-activated current which was used as a qualitative indicator of the intracellular calcium transient. 4. The ratio of first/steady state Ca-activated currents evoked by short action potentials was not modified. This may indicate that compound II does not influence the normal functioning of the sarcoplasmic reticulum stores. 5. The observations are consistent with the hypothesis that action potential prolongation by compound II reduces Ca2+ extrusion via the Na-Ca exchange. This in turn allows increased uptake of calcium into the sarcoplasmic reticulum stores so that more calcium is available for release by subsequent action potentials, leading to an increase in intracellular calcium transients and contractions. PMID- 8220920 TI - Amyloid arthritis associated with IgM kappa lymphoplasmacytoid lymphoma. AB - Amyloid arthritis is an uncommon cause of locomotor disease and may closely resemble RA. Macroglobulinaemia is rarely associated with amyloidosis and there has been only one report of amyloid arthritis in this setting, the patient having had Waldenstrom's macroglobulinaemia. We report the occurrence of amyloid joint disease in the course of an IgM kappa B-cell dyscrasia which evolved over 16 years to an overt lymphoplasmacytoid lymphoma. PMID- 8220921 TI - Glomerulonephritis in rheumatoid arthritis. AB - We present data on 10 patients with RA who developed glomerulonephritis which was not related to gold or penicillamine therapy. Although two of these patients had received gold this had been discontinued 13 and 18 yr before the diagnosis of glomerulonephritis. Seven patients presented with nephrotic syndrome and three with proteinuria and microscopic haematuria. Renal histology showed a membranous nephropathy (five patients), mesangial IgA nephropathy (two patients), focal segmental necrotizing glomerulonephritis (two patients) and focal segmental glomerulosclerosis (one patient). PMID- 8220922 TI - Re-evaluating the need for hospitalization following synovectomy using Yttrium-90 silicate. AB - In 51 patients treated with Yttrium-90 (Y-90) synovectomy for rheumatoid (inflammatory) arthritis (IA) and OA of the knee we found that decreased retained knee activity (RKA) and increased extra-articular activity in lymph nodes and liver are more likely to be found in IA than OA and following bilateral knee injections. Joint inflammation, as assessed by radionuclide blood pool scan but not by SF white cell count, correlates with decreased RKA and increased activity in lymph nodes. Intra-articular steroid had no significant effect on retention or extra-articular uptake. Strict hospital immobilization improves RKA of Y-90 in IA but not in OA. Y-90 synovectomy in OA shows good RKA and low extra-articular uptake. We recommend strict immobilization following Y-90 synovectomy, particularly in IA patients and/or those with high joint blood flow. PMID- 8220923 TI - Reactive arthritis associated with Bacillus Calmette-Guerin immunotherapy for carcinoma of the bladder: a report of two cases. PMID- 8220924 TI - Chronic polyarthritis due to Pseudomonas aeruginosa. AB - We present a 45-year-old male patient with chronic obstructive lung disease treated with low-dose corticosteroids, who developed a chronic septic polyarthritis due to Pseudomonas aeruginosa following a surgical wound infection. Due to the mild synovitis and the absence of systemic signs of infection the diagnosis was delayed for nearly 2 years and resulted in severe joint destruction. PMID- 8220925 TI - The Arthritis and Rheumatism Council guidelines for preparing a grant proposal for a clinical research project. PMID- 8220926 TI - Trichinosis mimicking polymyositis in a patient with human immunodeficiency virus infection. PMID- 8220927 TI - Increased secretion of norepinephrine after intravenous gamma globulin therapy in a patient with rheumatoid arthritis. PMID- 8220928 TI - Once bitten twice shy. PMID- 8220930 TI - Measures of pain and disease activity in rheumatoid arthritis. PMID- 8220929 TI - Anti-DNA antibodies in the primary antiphospholipid syndrome. PMID- 8220931 TI - MRI of the appendicular skeleton. PMID- 8220932 TI - Communication channels of the Arthritis and Rheumatism Council--the role of the Honorary Medical Secretaries. PMID- 8220933 TI - A new biochemical marker for joint injury. Analysis of YKL-40 in serum and synovial fluid. AB - We report the development of the first radioimmunoassay for YKL-40, a M(r) = 40 kDa protein which is secreted at high levels by human synovial cells and articular cartilage chondrocytes, and by the human osteosarcoma cell line MG63. This assay uses YKL-40 purified from the conditioned medium of MG63 cells as standard and tracer, and as antigen for immunizing rabbits. With this assay we have discovered high levels of YKL-40 antigen in serum and SF. The molecular weight of serum and SF YKL-40 is identical to purified YKL-40. To evaluate the possible utility of YKL-40 in the assessment of joint disease, we measured YKL-40 in serum and SF of 49 patients with various forms of inflammatory and degenerative joint disease and in the serum of 50 normal adults. The YKL-40 level in serum was significantly higher (P < 0.001) in the patients compared to the normal adults, but there was no difference in serum YKL-40 between the patients with inflammatory joint diseases and OA. The SF levels of YKL-40 were 15-fold higher than serum levels and there was a significant correlation (r = 0.55, P < 0.001) between YKL-40 concentration in SF and serum. Although the tissue distribution of YKL-40 secretion is presently unknown, these observations suggest that a major portion of serum YKL-40 in fact arises from the joint.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8220934 TI - Mechanical conditioning of articular cartilage to prevalent stresses. AB - The possible correlation between joint stresses and cartilage compressive modulus is examined. The stresses acting upon different areas of the joints and cartilage compressive modulus in these areas were obtained for 15 pairs of ipsilateral human ankle and knee autopsy joint specimens. It was found that the cartilage compressive modulus was significantly correlated with the mechanical stress (r = 0.889 at P < 0.02 level of significance) in such a manner that cartilage subjected to higher predominant stresses was significantly stiffer than that subjected to lower predominant stresses. This was true when comparing ankle with knee joints, and also when comparing different regions within one single joint. Such a correlation is significant in that it indicates that cartilage may well be conditioned mechanically by the prevalent stress it is subject to. However, this correlation of data obtained from autopsy specimens is a necessary, but not sufficient condition, for the above hypothesis to be true. It is, therefore, concluded that further work on animals is necessary. PMID- 8220935 TI - Immunoassay of platelet-derived growth factor in the plasma of patients with scleroderma. AB - It has been postulated that platelet-derived growth factor (PDGF) is responsible for the abnormal fibroblast proliferation observed in scleroderma. In one previous study, plasma samples from patients with scleroderma caused increased mitogenesis in cultured fibroblasts, suggesting that the pathogenesis of scleroderma is related to increased plasma PDGF concentrations. To test this hypothesis, we used a sensitive, monoclonal antibody-based ELISA to measure PDGF in the plasma of 12 scleroderma patients. A rigorous sampling protocol prevented false elevations in plasma PDGF levels from ex vivo platelet degranulation: beta thromboglobulin concentrations were measured in each plasma sample to monitor platelet lysis. Plasma PDGF concentrations in the scleroderma patients were not statistically different from those observed in age- and sex-matched normal controls, and patients with RA. While it is possible that changes in PDGF activity at a local level alter fibroblast function, we cannot conclude that elevated plasma concentrations of PDGF play a role in the pathogenesis of scleroderma. PMID- 8220936 TI - Avascular necrosis of the hip in systemic lupus erythematosus: the role of magnetic resonance imaging. AB - The value of magnetic resonance imaging (MRI) in the early diagnosis of avascular necrosis (AVN) of the hip in SLE was investigated. Twenty females with severe SLE were studied prospectively. Each underwent 6-monthly X-rays, technetium -99m (Tc 99m) pyrophosphate bone scans and MRI of the hips over a 3-yr period. AVN was diagnosed in five hips of three patients (15%) during the study period. It was confirmed histologically in three hips of two patients who underwent core decompression. Radiological evidence of AVN was present in two patients at diagnosis. One patient developed progressive radiological changes despite core decompression. Bone scintigraphy was abnormal at some stage in all three patients with AVN however failed to detect the early ischaemic stage of AVN. MRI was the most reliable investigation and was able to detect asymptomatic AVN prior to the appearance of radiological or scintigraphic abnormalities. PMID- 8220937 TI - A study of the early and late 99technetium scintigraphic images and their relationship to symptoms in osteoarthritis of the hands. AB - Thirty-five patients with OA of the hands had an early and late phase isotope bone scan performed at entry and 1 yr later. Simultaneous assessment of symptoms was made by a visual analogue pain score (VAS) and a tender joint articular index (AI) and comparisons were made between the clinical and scintigraphic findings. Sixty-five percent of joints were classed as positive in the late phase compared to 17% in the early phase scan. Thirteen per cent of joints were positive only in the early phase and 49% only in the late phase. There was no significant overall change in either phase of the scan in 1 yr. The pattern of positive joints showed considerable symmetry and wrist involvement. There was a high degree of correlation between AI and the late phase scan but none with the early phase scan. VAS showed no correlation with the late phase scan but did correlate significantly with the early phase scan at 1 yr. PMID- 8220938 TI - Effects of fish oil supplementation on non-steroidal anti-inflammatory drug requirement in patients with mild rheumatoid arthritis--a double-blind placebo controlled study. AB - Maxepa contains eicosapentaenoic acid (EPA) (171 mg/capsule) and docosahexaenoic acid (DHA) (114 mg/capsule). EPA acts as an alternative substrate to arachidonate, leading to the formation of the less proinflammatory prostaglandins ('3' series) and leukotrienes ('5' series). If Maxepa has anti-inflammatory properties it could be expected to reduce the requirement for NSAIDs in patients with RA. This has not been investigated nor has Maxepa therapy been studied over a full 1-yr period. Sixty-four patients with stable RA requiring NSAID therapy only were studied. Patients received either 10 Maxepa or air-filled placebo capsules per day for 12 months. All then received placebo capsules for a further 3 months. Patients were reviewed at 3-monthly intervals. NSAID requirement at entry visit for each patient was assigned as 100%. Patients were instructed to slowly reduce their NSAID dosage providing there was no worsening of their symptoms. Clinical and laboratory parameters of RA activity were also measured. There was a significant reduction in NSAID usage in patients on Maxepa when compared with placebo from month 3 [mean (95% C.I. for mean) requirement--71.1 (55.9-86.2)% and 89.7 (73.7-105.7)%, respectively]. This effect reached its maximum at month 12 [40.6 (24.5-56.6)% and 84.1 (62.7-105.5)%, respectively] and persisted to month 15 [44.7 (27.6-61.8)% and 85.8 (60.5-111.1)%, respectively] (P < 0.001, ANOVA). These patients were able to reduce their NSAID requirement without experiencing any deterioration in the clinical and laboratory parameters of RA activity. PMID- 8220939 TI - The prevention and healing of acute non-steroidal anti-inflammatory drug associated gastroduodenal mucosal damage by misoprostol. AB - This double-blind study assessed the acute development of NSAID-associated gastroduodenal (GD) damage and its prevention by misoprostol. Patients requiring chronic NSAID therapy were stratified into two groups depending on initial endoscopic appearance, Group I: normal (n = 223); Group II: non-ulcer lesions (n = 78). After 2 weeks of therapy with NSAID and either misoprostol 400-800 micrograms daily or placebo the incidence of severe mucosal damage (including ulcers) was significantly reduced by misoprostol (odds ratio; 95% CI). Group I: 4.52; 1.94, 10.51 (P = 0.018); Group II: 10.93; 1.09, 109.60 (P = 0.014); Groups I and II combined: 5.95; 3.23, 10.94 (P = 0.0003). Misoprostol exerted a significant protective effect against progression of minor to severe damage in Group II (P < 0.001). Endoscopic findings did not correlate significantly with gastrointestinal symptoms and misoprostol did not interfere with the NSAID efficacy. Significant GD damage occurs early in the course of NSAID treatment and misoprostol significantly reduces the incidence of such damage. PMID- 8220940 TI - Effect of a non-steroidal anti-inflammatory drug, naproxen, on faecal microbial flora. AB - Faecal Clostridium perfringens counts have been observed to be elevated in RA patients. The use of NSAIDs has been suggested as being responsible for this increase. To clarify the potential of NSAIDs to change faecal flora, 10 male volunteers were given naproxen 500 mg twice daily for 2 weeks in a randomized, placebo-controlled and double-blind study, and 10 other volunteers were given a placebo in tablets of identical appearance. Stool samples were collected and subjected to direct stool sample gas-liquid chromatography of bacterial fatty acids. The method has proved to be practical and sensitive in detecting overall changes in faecal flora. The samples were also cultured for Cl. perfringens. No significant change of faecal flora was observed by either method. The results show that naproxen given in doses and over a period in excess of the levels reported to increase intestinal permeability, does not change intestinal flora. PMID- 8220941 TI - The theory of reasoned action and cooperative behaviour: it takes two to use a condom. AB - The applicability of the Theories of Reasoned Action and Planned Behaviour to the cooperative behaviour of condom use were examined. Seventy-one male and 78 female students, all sexually active unmarried heterosexuals aged 17 to 21 years, gave information about their intentions for the next sexual encounter, as well as their attitude, subjective norm, expectancy-value attitude and subjective norm (including normative beliefs for their sexual partner), and their past behaviour with respect to condom use. After their next sexual encounter, they completed a questionnaire on their actual condom use. Results indicated that when behavioural conditions including the availability of a condom and an agreement with the partner to use it were satisfied, intention interacted with past behaviour to predict actual behaviour. These results imply that intentions which are consistent with past behaviour are stable enough to be carried out in the face of the interpersonal dynamics of a sexual encounter. Further, normative belief for the sexual partner had a direct influence on attitudes, subjective norm and intention. Neither the Theory of Reasoned Action nor the Theory of Planned Behaviour can fully explain these results, which point to the need for further theoretical inquiry into the dynamics of cooperative behaviour. PMID- 8220942 TI - The patello-femoral joint--a critical appraisal of its geometric assessment utilizing conventional axial radiography and computed arthro-tomography. AB - In the quest for treatable causes of anterior knee pain, plain film skyline views of the patello-femoral joint are requested, often in 30 degrees, 60 degrees and 90 degrees of flexion, to assess the functional relationships of the joint. Patello-femoral malalignment predisposes to recurrent subluxation and dislocation, articular cartilage damage and premature degenerative change. The aim of this study is to evaluate critically, by comparative assessment, the information provided by skyline views and axial computed arthro-tomography (CTA). Measurements of the patello-femoral angle (assesses patellar tilt), the congruence angle (assesses patellar lateralization) and the trochlear depth were made on Merchant's skyline views and on axial CTA on each of 50 symptomatic knees. Results are presented in graphic form with visual examples indicating a poor correlation between the two imaging methods. We conclude that skyline views are inaccurate and unsuitable primarily because they cannot be obtained in less than 30 degrees of flexion. We suggest that skyline views have no role to play in screening for maltracking as even florid examples must be missed, and would strongly urge that no surgery be performed on their basis alone as this would result in inappropriate operations. Although computed tomography is the preferred mode of assessing patello-femoral geometry, difficulties are still encountered and the ways of circumventing them are discussed. PMID- 8220943 TI - Computed tomographic cholangiography using spiral scanning and 3D image processing. AB - Volumetric computed tomography (CT) scans ("spiral CT") were performed after intravenous (i.v.) cholangiography followed by additional 3D surface reconstructions of gallbladder and biliary ducts. 34 patients were investigated prior to cholecystectomy. No allergic adverse reactions were observed. The scan time was 24 s. Contrast enhancement in the extrahepatic bile duct and gallbladder were measured. All CT image series were reviewed independently by four experienced physicians (two radiologists, two surgeons) and compared for quality with conventional cholangiography on a three-point scale. The average rating for the demonstration of the biliary tract was significantly better for spiral CT than for conventional cholangiography (p < 0.01). In all cases sufficient contrast was found in the common bile duct (mean 315 HU). 3D imaging was considered to be helpful for intraoperative orientation during laparoscopic surgery. PMID- 8220944 TI - Elevation of the larynx on normal and abnormal cineradiogram. AB - The relationship between bolus volume (2.5, 5, 10 and 20 ml) and larynx elevation during swallowing was assessed in 10 non-dysphagic and 10 dysphagic individuals without pharyngeal dysfunction. Laryngeal elevation in different types of pharyngeal dysfunction was assessed in 60 non-dysphagic and 75 dysphagic patients. All subjects were examined with liquid barium and cineradiography at 50 frames/s. The laryngeal elevation was measured at the moment when the bolus reached the level of the valleculae and at maximum elevation. Elevation of the larynx, both the initial and maximal, was not influenced by sex, age or presence of dysphagia. Elevation of the larynx at the moment when the bolus reached the valleculae, when expressed in per cent of maximum elevation, was lower with 10 and 20 ml bolus volumes compared with 2.5 ml bolus volume (p < 0.05) in the 10 dysphagic individuals. Pharyngeal dysfunction was associated with significantly lower initial elevation of the larynx, at the moment when the bolus reached the level of the valleculae, although the maximal laryngeal elevation was normal. Initial elevation was 30% lower (p = 0.03) in patients with aspiration of bolus material into the trachea, 22% lower (p = 0.007) in those with defective closure of the laryngeal vestibule without aspiration and 16% lower (p = 0.06) in those with incoordination of the cricopharyngeal muscle compared with patients without dysfunction. PMID- 8220945 TI - The histological response of the lungs of rats to potentially suitable water soluble bronchographic contrast agents iotrolan (a non-ionic dimer) and iopamidol (a non-ionic monomer). AB - The recent need for selective bronchography during bronchoscopy in certain patients, together with paediatric indications for tracheo-bronchography, raises the question as to which contrast medium is both safe and efficient. The purpose of the present experimental study was to define the tissue reaction caused by iotrolan and iopamidol in the bronchi and lungs of rats. 60 animals used in this study were divided into five groups receiving iotrolan 300, iopamidol 370, iopamidol 150, physiological saline and anaesthetic only, the last two acting as control groups. Statistically there was no significant difference in the histological reaction between iotrolan 300, iopamidol 370 and iopamidol 150 compared with the control groups, although iotrolan 300 appeared to result in the least tissue reaction. With its adequately high iodine concentration, low osmolality and relatively high viscosity, iotrolan 300 would appear to be a suitable contrast medium for bronchography. PMID- 8220946 TI - The effects of radiographic contrast media on leucocyte orientation. AB - The effects of radiographic contrast media (RCM) on leucocyte orientation in vitro were studied using a Zigmond chamber. Leucocyte orientation was assessed following exposure of the leucocytes to iotrolan, iohexol, ioxaglate and diatrizoate. The RCM used were diluted to a concentration similar to that obtained in vivo during routine angiography. At this concentration there was a significant reduction in leucocyte orientation for all RCM investigated but the effect was more pronounced in the monomeric than the dimeric RCM. The results may have significance when deciding which radiographic contrast medium to use in selected patients, particularly those who are immunosuppressed or septicaemic. PMID- 8220947 TI - The management of stroke during intraarterial thrombolysis. AB - Intraarterial thrombolytic therapy is being increasingly used in the treatment of acute limb ischaemia. When strokes occur during thrombolytic therapy treatment is usually stopped because of the suspicion of cerebral haemorrhage. If the stroke is not haemorrhagic then stopping the treatment jeopardizes the ischaemic limb. If the stroke is proven to be non-haemorrhagic on computed tomography (CT) then the thrombolytic therapy should be recommenced. We present three case histories of patients who suffered a stroke during thrombolytic therapy over a 30-month period, in whom CT confirmed non-haemorrhagic stroke and we suggest that managing the ischaemic limb with anticoagulant and thrombolytic therapy can be safe in the early period following the stroke. We also discuss the probable aetiology of the strokes and whether cardiac echocardiography should be performed in these patients. PMID- 8220948 TI - T1 rho dispersion imaging of diseased muscle tissue. AB - T1 rho dispersion, or the frequency dependence of T1 relaxation in the rotating frame, was used for in vivo muscle tissue characterization in 13 patients with primary skeletal muscle disease and in eight normal subjects for comparison. T1 rho dispersion measurements represent a new approach to magnetic resonance tissue characterization, possibly reflecting the macromolecular constituents of tissue. A definite, statistically significant, difference was found between the relative T1 rho dispersion values of normal and diseased muscle tissue. T1 rho dispersion measurements and images may increase the accuracy of identification of diseased muscles. Early identification of affected muscles is important for accurate diagnosis by muscle biopsy. PMID- 8220949 TI - Gastrointestinal manifestations in the Hallopeau-Siemens variant of recessive dystrophic epidermolysis bullosa. AB - Epidermolysis bullosa encompasses a group of rare disorders typified by blister formation following minor trauma to the skin. Gastrointestinal tract involvement may occur in the extremely rare recessive dystrophic variants. The gastrointestinal manifestations present in 25 patients with the Hallopeau-Siemens variant of recessive dystrophic epidermolysis bullosa are reviewed. In the oesophagus both anatomical and motility abnormalities were observed. Features seen included a generally shortened oesophagus, strictures including those resembling webs, hiatus herniae, decreased peristalsis, oesophageal atony and pseudodiverticulum formation. These patients also had faecal impaction. PMID- 8220950 TI - Stab wounds of the heart: two new signs of pneumopericardium. AB - The diagnosis of a small traumatic pneumopericardium may be difficult and differentiation from a left medial pneumothorax or pneumomediastinum is important. Two new signs which only occur in pneumopericardium are described: the "transverse band of air" sign on the frontal radiograph representing air in the transverse sinus of the pericardium, and the "triangle of air" sign as noted on the lateral radiograph. If there is still doubt, a left-side-down decubitus radiograph will establish the diagnosis. PMID- 8220951 TI - Changes in the radiation treatment of cancer of the anus in Glasgow. AB - The survival and complication rate of two groups of patients with carcinoma of anus were compared. The first group of 20 patients were treated between 1974 and 1983 by double plane implantation using radium or equivalent caesium needles and supplementary external beam radiotherapy to groin nodes if present. The second group of 26 patients were treated between 1984 and 1990 by external beam radiotherapy followed by a jig implant using afterloading iridium needles. 5-year survival was superior in the second group (73% vs. 40%) with a lower incidence of radiation-induced complications. PMID- 8220952 TI - Digital storage phosphor radiography for treatment verification in radiotherapy. AB - The potential of digital storage phosphor radiography (SR) to improve image quality of portal radiographs is evaluated. Conventional film radiographs (FR) and corresponding SR verification images of an anthropomorphic phantom and various irradiation ports of patients were obtained with high-energy photon beams. For both techniques conventional films and storage phosphor screens were placed into a cassette with steel screens. Images were evaluated according to contrast and spatial resolution, delineation of anatomical structures, position of shielding blocks and accuracy of field alignments. Evaluation of 33 pairs of SR and FR portal images yielded a superior contrast resolution of SR in 47% (contrast air-soft tissue) and 37% (bone-soft tissue). Thus SR allows quick and easy detectability of anatomical structures as well as a better definition of block positions and field alignments. Shorter exposure times for computed images may result in a reduction of motion artefacts. SR images are indispensable in modern radiation therapy units, as they are instantly available in a computerized network for further image processing and analysis. PMID- 8220953 TI - Is there a danger in delaying radiotherapy in childhood medulloblastoma? AB - Approximately 45-50% of children with medulloblastoma are cured by conventional surgery and radiotherapy, but survivors may face severe late neuropsychological toxicity. Studies showing good partial responses to platinum-based chemotherapy in relapsed patients and the theoretical possibility of a therapeutic window immediately after surgery have prompted neoadjuvant treatment studies which are ongoing. However, the absolute benefit of chemotherapy for the treatment of medulloblastoma in childhood is, as yet, not proven. There is a danger that chemotherapy may simply delay radiotherapy, and in so doing reduce the radiological impact of this known effective treatment. We report four children with medulloblastoma presenting consecutively to this unit over a 6-month period, whose management was problematic because of either failure to respond to neoadjuvant chemotherapy or their very young age. These cases are discussed in the light of the current literature and future treatment strategies that must seek to improve the therapeutic ratio of multimodality therapy. PMID- 8220954 TI - Reduced bone mineral density in long-term survivors of medulloblastoma. AB - Bone mineral density (BMD) reaches a peak at approximately 30 years of age, and may be influenced by radiotherapy before completion of skeletal maturation. Regional BMD has been measured using dual energy X-Ray absorptiometry (DEXA) in adults following craniospinal irradiation for medulloblastoma between ages 4 and 19 years, receiving doses of 3500-4000 cGy to the brain and spinal cord. Lumbar spine (LS) and was failure to achieve normal adult BMD at both LS and FN, with a mean reduction at LS of 12.1% +/- 2.4% (p < 0.01) and a mean reduction at FN of 14.3% +/- 3.4% (p < 0.01). The mean body mass index (BMI) was also less than that of a standard population (21.8 +/- 1.5), as were mean standing and sitting heights. No relationship was found between reduction in BMD at either site and age at irradiation, time elapsed since irradiation or BMI at time of scanning. Biochemical and endocrine markers including corrected calcium, alkaline phosphatase, sex hormones and IGF-1 were normal in all seven patients. The reduction in BMD outside the irradiated area suggests that indirect factors may be important in this effect. PMID- 8220955 TI - Clinical high resolution skeletal single photon emission tomography using a triple-headed gamma camera. AB - Planar skeletal scintigraphy has become established as a standard diagnostic test performed within the nuclear medicine department. Since the 1970s good quality images have been produced using an Anger gamma camera and 99Tcm-labelled diphosphonates. Single photon emission tomography (SPET) has improved the sensitivity of detection and the ability to localize bony pathology, particularly benign bone disease in the spine. Recently multi-detector gamma cameras dedicated to SPET have become available. One such system, the Toshiba GCA-9300A, has been used to perform routine clinical skeletal SPET in 81 patients. Good quality images have been obtained using an 8 min acquisition in the axial skeleton and a 16 min acquisition protocol in the peripheral skeleton. Multiple sites can be tomographed in the same patient during the same examination using two or more 8 min acquisitions. Such a multi-detector gamma camera offers advantages over the standard single-headed rotating camera for skeletal SPET in terms of both imaging time and image quality. A cost analysis was performed which demonstrated that the additional cost of purchasing such a multidector gamma camera was less than 30.00 pounds per SPET study. PMID- 8220956 TI - Radiation doses to paediatric patients undergoing less common radiological procedures involving fluoroscopy. AB - A semi-automated dosimetry survey has been undertaken to monitor the radiation doses to patients in a paediatric fluoroscopy room. The doses were assessed using large area Diamentor ionization chambers to measure dose x area product, and by means of thermoluminescent dosemeters attached to the patient's skin in various places. Details of patients and examinations were entered into a computer, all information being stored on a data base. Many radiological examinations have been monitored, including renal and jejunal biopsies, nephrostograms, loopograms, tracheograms and fistulograms. Patients included in the study have been divided into three age bands for dose assessment. Radiation doses for different types of examination have been compared, including more common examinations for which data have been previously presented. PMID- 8220957 TI - Technical note: physical evaluation of recent Kodak films for mammography. AB - Four Kodak films for mammography were evaluated for image quality and dose. Processing times and storage effects have also been evaluated. PMID- 8220958 TI - Technical note: the implementation of patient position correction using a megavoltage imaging device on a linear accelerator. AB - The problem of using information from the analysis of megavoltage images to adjust patient set-up has been addressed. In the case of rotational corrections it has been assumed that the treatment head is to be adjusted, although for gantry angles of 0 degree and 180 degrees couch rotation may be used. In the case of translational shifts adjustment of the collimator jaws or of the couch have both been considered for arbitrary combinations of couch and gantry angle. For couch movement the case has been considered where it is desirable to minimize both the number of parameters to be adjusted and also the magnitude of the change in the patient's position. Values obtained for frequently used set-up parameters have been presented. Adjustment of the treatment couch positioning is the most desirable option, as this should bring the patient closer to the correct position for subsequent treatment fields. However, rotational errors are not correctable for all gantry angles and furthermore the collimator settings may be set more accurately than those of the treatment couch. Hence, in some cases, adjustment of the collimation system may be desirable or necessary. The formulae given in Equations (13) to (18) are currently being used in an intervention study to correct patient set-up during the course of a treatment fraction. PMID- 8220959 TI - Case report: accessory lobe of the liver mimicking lesser omental lymphadenopathy. AB - An accessory lobe of the liver is an uncommon occurrence and often an incidental finding. We present the case of a middle-aged man whose upper abdominal pain was relieved following removal of an accessory lobe. His preoperative imaging had led us to believe that he had lesser omental lymphadenopathy. We draw attention to the fact that this congenital abnormality can cause diagnostic confusion. PMID- 8220960 TI - Case report: emphysematous pyelonephritis caused by clostridium septicum and complicated by a mycotic aneurysm. PMID- 8220961 TI - Case report: staghorn calculus complicated by psoas abscess presenting as flank mass in a teenager. PMID- 8220962 TI - Case of the month: computed tomography in the diagnosis of pyrexia of unknown origin. PMID- 8220963 TI - Squamous cell carcinoma of the nail bed. PMID- 8220964 TI - Assessment of X-ray field alignment in mammography. PMID- 8220965 TI - Complications of intraarterial digital subtraction angiography in patients investigated for cerebral vascular disease. AB - 395 patients (236 males, mean age 55.6 years: 159 females, mean age 52.2 years) with suspected transient ischaemic attacks or previous strokes underwent intraarterial digital subtraction angiography (IADSA) over a 3-year period ending in March 1991. All procedures were performed via the femoral approach and the majority consisted of arch studies followed by selective catheterization. 253 (64.1%) of the patients had extracranial vascular disease confirmed at angiography. A retrospective analysis of the patients' records was made to extract all possible complications. Complications were defined as any untoward symptoms or signs occurring within 48 h and which could have been related to the angiogram. Neurological complications occurred in 15 (3.89%) patients. 10 (2.5%) patients had transient complications which resolved completely within 24 h. In three (0.8%) patients the neurological deficit was reversible, recovering fully within 6 days. Two (0.52%) patients were left with residual disability from stroke at 10 days. The permanent neurological complication rate is in the lower range of the rates recorded in previous conventional angiographic studies. We conclude that IADSA is a relatively safe and reliable form of investigation in patients with suspected cerebral vascular disease. PMID- 8220966 TI - Ultrasound diagnosis of splenic lymphoma: ROC analysis of multidimensional splenic indices. AB - To assess the efficacy of splenic size in the sonographic diagnosis of lymphomatous involvement of the spleen, the authors studied 31 patients with splenic lymphoma (Hodgkin's disease, 17 and non-Hodgkin's lymphoma, 14) and 218 individuals without evidence of splenic disease. All subjects were studied with both a linear and a sector transducer. The longitudinal, transverse and diagonal diameters of the spleen were measured, and two- and three-dimensional splenic indices were calculated. The analysis of the diagnostic performance of these criteria, compared by means of receiver-operating characteristic (ROC) curves, revealed that diagnosis of splenic involvement by malignant lymphoma was considerably more reliable with a sector than with a linear scanner. If the longitudinal diameter was measured with a sector scanner, sensitivities were 66% and 74%, at specificities of 95% and 90% respectively (cut-off points: 12.5 cm and 11.3 cm, respectively). For the sector scanner, there was no advantage in using other diameters or multidimensional indices. Additional ROC analysis of recently published data indicating excellent discrimination capacity of a computed tomography index revealed that these results were largely owing to patient selection. In contrast, our data suggest that the potential of current non-invasive assessment of splenic lymphoma is limited. However, ultrasound may eventually help to eliminate staging laparotomy in selected cases, e.g. in patients with low risk of abdominal disease and with increased surgical risk. PMID- 8220967 TI - Quality assurance in obstetric ultrasound. AB - Ultrasound examination is a routine element of antenatal care, and an accurate and reliable ultrasound service is essential for confident patient management. Qualitative findings, e.g. fetal normality and placental site, may be confirmed at delivery, but this form of audit is not suitable for fetal measurements owing to the variation in gestation at delivery and the complexity of neonatal assessment. Our aim was to develop a simple audit method applied a short time after the ultrasound scan. The method was based on assessment of measured images against measurement criteria which are clearly defined in the literature, results for each criterion being recorded in a spreadsheet. Two main forms of report were generated, the first showing overall achievement of satisfactory measurements for each sonographer, the second providing graphical information to show which criteria required greater attention by individual sonographers. Over several phases of audit problems of quality recognition and technical skill were isolated, graphical reports were used to guide tuition and the levels of performance were improved. The method itself and the results satisfy managers at all levels that standards are in place and are being maintained. PMID- 8220968 TI - A double blind study to evaluate the tolerability of gadodiamide injection and its effect on renal function in patients undergoing cerebral magnetic resonance imaging. AB - Gadodiamide injection was administered intravenously to 28 patients with cancer undergoing cerebral magnetic resonance imaging (MRI). Two parallel groups were used to evaluate the safety of single doses of 0.1 and 0.3 mmol per kilogram body weight (kgbw). Adverse events, vital signs, blood chemistry, haematology and urinalysis were the principal measures of safety. Four patients, all in the 0.1 mmol kgbw-1 group, experienced a total of six adverse events. No adverse events were reported in the 0.3 mmol kgbw-1 group. No clinically significant changes in blood chemistry, haematology or urinalysis occurred. No significant changes in renal tubular function or glomerular filtration rate were observed after injection at either dose. Overall, this study suggests that gadodiamide injection is a safe and effective contrast medium for use in patients undergoing cerebral MRI at both the 0.1 and 0.3 mmol kgbw-1 doses. PMID- 8220969 TI - Does the Whitaker test add to antegrade pyelography in the investigation of collecting system dilatation in renal allografts? AB - Significant pelvicalyceal dilatation in renal allografts is currently investigated by antegrade pyelography. However, the clinical significance of a radiologically demonstrated narrowing of the ureter is unclear. Over a 21-month period 26 of 155 renal allografts with pelvicalyceal dilatation were investigated by antegrade pyelography. In eight allografts no ureteric stenosis could be identified. Two grafts were shown to have ureteric necrosis and required surgical intervention and 16 of the other grafts appeared to have a ureteric stenosis. 15 of the 16 allografts with radiological ureteric stenosis underwent a concurrent pressure flow study to assess the functional relevance of the ureteric narrowing. As shown by a pressure rise of > 7 mmHg at a perfusion rate of 10 ml min-1, 11 of the 15 grafts were functionally obstructed and were treated by a nephrostomy catheter followed by antegrade insertion of a ureteric stent. The four grafts with a negative pressure flow study were subsequently shown on biopsy to have rejection. The diagnosis of allograft rejection was also confirmed by biopsy in seven of the eight allografts without a radiological ureteric stenosis. The last of the eight allografts was found to be cyclosporin toxic. Pelvicalyceal dilatation of renal allografts is appropriately investigated by antegrade pyelography in combination with a pressure flow study which identifies those grafts with mechanical obstruction. PMID- 8220970 TI - Quantification of image persistence in a digital angiography system. AB - Image persistence, as a characteristic of video imaging systems affecting the quality of fast moving fluoroscopic images, is shown to vary considerably. A simple quantitative method for measuring image persistence in a digital angiography system is presented, together with a series of image intensifier exposure-response curves. For the Saticon tube, used with the Siemens 3VA Digitron, it was found that persistence increased for low exposure rates and may increase to 31% at a 120 ms interval. In addition, a sharp increase in image persistence, from 8.3% to 33%, was observed within 18 months from installation of the system. PMID- 8220971 TI - Fat suppressed magnetic resonance imaging at 0.5 T using binomial radiofrequency pulses. AB - Fat suppressed MR imaging can be achieved by selectively saturating lipid protons, just before applying a conventional spin-echo rf pulse sequence. The difference in the Larmor frequency between fat and water protons is only 3.5 ppm, so that the frequency response of the suppressing pulse, or pulses, has to be carefully designed. The choice of suitable binomial and modified trains of hard rf pulses was investigated by computer simulation, phantom testing and T1 weighted spin-echo imaging of a normal volunteer's optic nerve at 0.5 T. PMID- 8220972 TI - Evolution of diagnostic radiology in a big hospital during a 5 year period, and the derived collective dose. AB - An analysis is presented of the trends in the annual number of radiological examinations and in the average effective dose equivalent for each type of examination in a big Spanish hospital. Annual frequencies for each type of examination, annual average effective dose equivalent values for each study group, and the contribution of each examination group to the collective dose are presented. Also, sex and age distributions for several important examinations are given, and their impact on the collective dose is reviewed. PMID- 8220973 TI - Radiation exposure to the hands of orthopaedic surgeons during procedures under fluoroscopic X-ray control. AB - The hands of the surgeon are most likely to be directly exposed to ionizing radiation during fluoroscopic screening in the orthopaedic theatre. There is however little information available on the level of exposure to radiation during the normal working pattern of individual surgeons. The purpose of this study was to directly measure the radiation exposure to the hands during fluoroscopic screening in a series of consecutive cases over a month in order to establish whether these staff need to be designated classified persons, and if not, whether they need to be routinely monitored. Extremity monitoring was carried out using thermoluminescent dosimeters. The dosimeter was secured to the operating surgeon's dominant index finger. 44 procedures were carried out by nine different surgeons. The total radiation dose received per surgeon ranged from 48-2329 microSv. In 80% of procedures the dose of radiation to the surgeon's hand was less than 100 microSv. The extrapolated annual dose, even for the surgeon with the highest radiation exposure, was well below the annual dose limit for extremities of 500 mSv per year recommended by the International Commission on Radiological Protection, and embodied in the Ionizing Radiations Regulations 1985. Despite the relatively low doses of radiation received by surgeons in this study, occupational exposure to all personnel should be kept to the lowest practicable levels, and a review of procedures, including dose measurements, from time to time is advised. PMID- 8220974 TI - Dual X-ray absorptiometry: a comparison between fan beam and pencil beam scans. AB - The recent introduction of dual X-ray absorptiometry (DXA) systems with fan beam instead of conventional pencil beam scanning geometry represents a significant technical advance in bone densitometry. This report describes phantom and in-vivo studies of the effect of the change in beam configuration on DXA measurements. Fan beam and pencil beam measurements acquired on one of the new generation scanners, the Hologic QDR-2000, were compared with scans performed on an earlier pencil beam model, the Hologic QDR-1000. The variation with height above the scanning table of fan beam measurements of an anthropomorphic spine phantom were: bone mineral content (BMC): -3.1% cm-1; projected area (AREA): -2.8% cm-1; bone mineral density (BMD): -0.2% cm-1. For pencil beam scans the magnitude of height variation was less than 0.1% cm-1. QDR-2000 fan and pencil beam scan results for 20 volunteers correlated closely with QDR-1000 pencil beam data (r = 0.966 0.998). For BMD measurements on the spine and hip, differences between fan and pencil beam data were consistent with the errors expected from measurement precision. For AREA and BMC data, however, larger differences were observed with individual deviations which correlated with body habitus of the subjects. Although the change from pencil to fan beam geometry significantly affected AREA and BMC data, the effect on the clinically more important BMD measurements was negligibly small. PMID- 8220975 TI - The 62 MeV proton beam for the treatment of ocular melanoma at Clatterbridge. AB - A second treatment room and beam line has been constructed at the Cyclotron Unit at Clatterbridge for the purpose of using 62 MeV protons for the treatment of ocular melanoma. A uniform beam is produced by a double foil scattering system. The initial Bragg peak is spread across the target volume by the use of beam modulators. These are rotating four-vaned stepped absorbers made from Perspex. Two beam lines can be configured with different positions of modulators and range limiters. The first has a maximum penetration of 31.9 +/- 0.2 mm in water and the second a penetration of 31.2 +/- 0.2 mm. The second configuration has the advantage of less variation in beam penumbra, with a typical value of 1.7 +/- 0.1 mm for the 90% to 10% decrement lines. The patients are treated with individually shaped collimators. Beam output varies by less than 2% over the range of collimator areas used. The resulting whole-body dose equivalent to patient has also been assessed. In the first three years of operation over 250 patients have been treated. PMID- 8220976 TI - Radiation risks to personnel and public during the treatment of malignant glioma using interstitial brachytherapy. AB - 125I seeds are used in brachytherapy for the treatment of malignant gliomas. The use of such radioactive sources is associated with a certain radiological hazard to both personnel and members of the public. This hazard should be quantified so that the ALARA principle of radiological protection may be implemented satisfactorily. A study was undertaken to derive isodose rate contours in the vicinity of an anthropomorphic phantom with 125I seeds positioned at typical tumour sites within the cranial cavity. These contours are illustrated for seed positions appropriate to deep and superficial temporal tumours. Results indicate that the annual doses to personnel and public should not exceed those recommended by the International Commission on Radiological Protection. PMID- 8220977 TI - The detection and modification of the hypoxic fraction in quiescent cell populations in murine solid tumours. AB - Mice bearing SCC VII or EMT6/KU tumours were irradiated after receiving 10 injections of 5-bromo-2'-deoxyuridine (BUdR) to label all proliferating tumour cells, and the tumours were then excised and trypsinized. The tumour cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker), and the micronucleus (MN) frequency in cells without BUdR labelling was determined using immunofluorescence staining to BUdR. This MN frequency was then used to calculate the surviving fraction of unlabelled cells from the regression line for the relation between MN frequency and the surviving fraction of all tumour cells. Thus a cell survival curve could be determined for cells not labelled by BUdR, which can be regarded as quiescent tumour cells for all practical purposes. Assays performed immediately after irradiation of both normally aerated and hypoxic tumours showed that quiescent cells contained higher hypoxic fractions than the tumour cells as a whole. Furthermore, administration of nicotinamide before irradiation or the placement of mice in a circulating carbogen (95% O2, 5% CO2) chamber for 30 min before and during irradiation altered the acutely and chronically hypoxic fractions of the proliferating and quiescent tumour cell populations in a way which depended on the tumour system. Combined nicotinamide and carbogen therapy was shown to have a large potential to sensitize cells to low-dose radiation in vivo. In addition, this assay method appears to be useful for determining the size of the hypoxic fraction of quiescent tumour cells in murine solid tumours. PMID- 8220978 TI - Technical note: a reference frame designed to use external numeric references during simulation for tumours located in the head. PMID- 8220979 TI - Technical note: digital mammography--the comparative evaluation of film digitizers. PMID- 8220980 TI - Technical note: a re-evaluation of magnetic resonance imaging damping loops. PMID- 8220981 TI - Case report: primary intrathoracic rhabdomyosarcoma: a rare childhood malignancy. AB - Primary intrathoracic rhabdomyosarcoma is a rare tumour in childhood. Three cases are presented and the radiological findings and clinical course are reviewed. The radiological manifestations are varied but a rapidly growing soft tissue mass with compression of adjacent structures is the most common. A rare association with an underlying congenital pulmonary cyst is described. The prognosis is worse than for rhabdomyosarcoma at other sites, with a predisposition for cerebral metastases. PMID- 8220982 TI - Case report: an unusual cause of epistaxis: non-traumatic intracavernous carotid aneurysm. A case report with 12 year follow-up and review of the literature. AB - Intracavernous carotid aneurysms are uncommon. We report the natural history and radiological appearances of a giant, non-traumatic, intracavernous carotid aneurysm which extended through the skull base to the anterior nares and caused epistaxis. The magnetic resonance imaging appearances of such an aneurysm have not been previously described. The importance of correct diagnosis is discussed. PMID- 8220983 TI - Case report: infected false aneurysm at the site of an iliac stent. AB - False aneurysm formation at the site of iliac artery stent placement is an uncommon but serious complication of the procedure. We report a case of infected false aneurysm at the site of an iliac stent, complicated by renal failure. PMID- 8220984 TI - Case of the month: interesting ileus. PMID- 8220985 TI - Incidentally discovered solid renal masses: what are they? AB - In 36 of 99 consecutive patients operated on for the presumptive diagnosis of renal carcinoma the tumour was discovered accidentally during investigations for a variety of complaints. CT and ultrasound scans were responsible for discovery of the tumour in 24 of the 36 patients. Renal carcinoma accounted for 92% of incidentally discovered solid renal masses. Of 33 incidentally diagnosed renal carcinomas 27 were stage 1. An incidentally discovered solid renal mass should be regarded as a low stage renal carcinoma until proven otherwise. PMID- 8220986 TI - Severe sepsis following percutaneous or endoscopic procedures for urinary tract stones. AB - Nine cases of severe sepsis following percutaneous or endoscopic procedures for upper urinary tract stones are reported. The mortality rate was 66%. Despite the fact that approximately 700 procedures were carried out in males and females in roughly equal proportions, a striking but inexplicable feature was that all 9 patients in the study group were female. Severe systemic sepsis has a high mortality rate and any procedure that may put patients at risk of this complication should not be undertaken lightly (and certainly not as an out patient procedure). Recovery is possible with a high index of suspicion, early intervention and intensive treatment. PMID- 8220987 TI - The ureter in vitro: normal motility and response to urinary pathogens. AB - The effects of bacteria on in vitro ureteric contractility were studied, using a model which allowed selective exposure of organisms to the ureteric mucosa and smooth muscle, respectively. A cannula attached to a pressure transducer was ligated into the proximal lumen of 2.5-cm segments of canine ureter. The distal ureter was ligated to form a closed pressure monitored system, and the segment suspended in a 20-ml organ bath containing Krebs Henseleit buffer at physiological pH and temperature. Following onset of spontaneous activity, broths of Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa and Staphylococcus aureus were added to either the buffer solution or ureteric lumen in doses of > 10(6) organisms/ml. Experiments were repeated using heat-killed organisms, bacterial filtrates and E. coli endotoxin. Ureteric contractility was stimulated by organisms added to the buffer medium, but reversibly inhibited by bacteria placed in the ureteric lumen. Heat-killed organisms, endotoxin and live filtrates had no effect on normal motility when exposed to either the ureteric mucosa or muscularis respectively. These findings reflect the conflicting changes in ureteric motility seen in vivo when bacteria are administered systemically or directly into the ureteric lumen. PMID- 8220988 TI - Ureteric peristalsis studies in loin pain and haematuria syndrome: another diagnostic disappointment. AB - Although not a cause of progressive renal damage, loin pain and haematuria syndrome is nevertheless associated with significant morbidity. The management of pain often presents a formidable problem to urologists, physicians and general practitioners. An earlier study noted hold-up of urine in the renal pelvis and implicated this in the pathogenesis of pain. In this study we have failed to demonstrate an excess incidence of disordered urinary peristalsis in loin pain and haematuria syndrome. PMID- 8220989 TI - Metabolic appraisal of the effects of dietary modification on hypersensitive bladder symptoms. AB - The concentration of certain metabolites and amino acids appears to be changed in patients with the painful bladder syndrome interstitial cystitis. A study of the metabolism of the arylalkylamines (tryptophan, tyrosine, tyramine, phenylalanine) was carried out in 250 patients (237 females, 13 males), revealing an inability to synthesise normal amounts of serotonin and MHPG, a noradrenaline metabolite. Furthermore, the absence of ammonia and tryptophan in urine confirmed the presence of a membrane leak. Dietary restriction lessened the symptoms but did not alter specific abnormalities in dopamine metabolism. Dietary management offers a cost-effective therapeutic approach. PMID- 8220990 TI - Response of the human neurogenic bladder to KCl, carbachol, ATP and CaCl2. AB - The in vitro pharmacological responses of the human neurogenic bladder to KCl, carbachol, ATP and CaCl2 have been analysed. The contractility (contractile strength and ED50) of neurogenic bladders was significantly increased when treated with carbachol, ATP and CaCl2. In contrast, there was no apparent difference in the responsiveness of neurogenic bladders when treated with KCl. There was no apparent correlation between pharmacological responsiveness and clinical parameters, such as gender, age or cystometric data, in the neurogenic bladders. PMID- 8220991 TI - Search for mycobacteria in interstitial cystitis using mycobacteria-specific DNA probes with signal amplification by polymerase chain reaction. AB - The aetiology of interstitial cystitis is not known. Various infective agents have been postulated and although recognised as perpetrators of chronic inflammatory conditions, mycobacteria have never been satisfactorily excluded from interstitial cystitis. If present in interstitial cystitis tissue, mycobacteria exist either in very small numbers or in forms which contemporary staining techniques fail to recognise. We used a polymerase chain reaction with mycobacteria-specific DNA probes and found no evidence of mycobacterial involvement in 8 cases of proven interstitial cystitis. PMID- 8220992 TI - Urinary fibronectin in diagnosis and follow-up of patients with urinary bladder cancer. AB - The levels of fibronectin in urine from 106 patients with urinary bladder cancer, from 13 patients with benign urological disease and from 24 healthy control individuals were determined by an enzyme-linked immunosorbent assay (ELISA). The fibronectin levels in urine from patients with bladder cancer were higher than in patients with benign urothelial disease and in healthy controls. In 9 patients with bladder cancer, sampling was done both pre- and post-operatively. In these cases the fibronectin levels after operation were significantly lower than they had been before. Among 14 patients treated with BCG intravesically for superficial bladder tumours, those with complete remission of disease had less urinary fibronectin than those who did not respond to treatment. The data suggest that urinary fibronectin may be a useful marker for detecting urinary bladder cancer and for selecting patients for BCG treatment. PMID- 8220993 TI - Post-operative retention associated with acute prostatic infarction. AB - Twenty-one patients with post-operative retention following unassociated surgery and requiring transurethral resection of the prostate were compared with patients with acute retention (control group). Histological evidence of acute prostatic infarction was significantly increased in the post-operative retention group. Prolonged operative hypotension was associated with acute prostatic infarction, as were smoking and pre-existing cardiovascular disease. PMID- 8220994 TI - Evolving experience with radical prostatectomy. AB - Thirty-six radical prostatectomies were performed over an 8-year period; 25 suitable patients (70%) presented with symptoms of bladder outflow obstruction. In 15 cases (44%), initial digital rectal examination was not indicative of malignancy. The primary tumour was understaged pre-operatively in 17 patients (48%). In 14 cases (41%) the pre-operative biopsy grade was different from the grade assigned to the tumour following radical prostatectomy. Radical prostatectomy is being performed with increasing frequency: trends in morbidity have been identified. PMID- 8220995 TI - Epinephrine reduces the severity of catheter-induced urethral inflammation by action at the alpha 2-adrenoceptors. AB - We have studied the contribution of epinephrine to experimentally induced urethral inflammation in the rat. Inflammation was induced by inserting latex strips into the urethra. The effects of various experimental procedures were assessed according to a 4-point scale based on histological findings. The results showed that 0.5 mg/kg epinephrine decreased the severity of catheter-induced urethral inflammation. This effect was blocked by the alpha 2-adrenergic agonist, yohimbine, but not by alpha 1 (prazosin), beta 1 (M32 MTC), or beta 2 (butoxamine) antagonists. The results suggest that the suppressive effect of epinephrine is mediated by action at the alpha 2 adrenergic receptor. PMID- 8220996 TI - Electromyography of cavernous smooth muscle during flaccidity: evaluation of technique and normal values. AB - Objective evaluation of the penile innervation in impotent patients is mostly restricted to examination of the somatic pudendal pathways. These tests provide little information on the pelvic-cavernous autonomic innervation of the corporeal bodies. Electromyography of the flaccid penile smooth muscle is a reproducible and non-invasive method of evaluating these autonomic pathways and the status of intrinsic smooth muscle. Examination techniques and normal values have been studied in 15 young and potent volunteers. Recordings in 13 patients with neuropathology and 57 impotent patients are discussed. PMID- 8220997 TI - Renal transplantation in young boys with posterior urethral valves: preliminary report. AB - Seven boys (mean age 38 months) with posterior urethral valves underwent renal transplantation between June 1988 and August 1991. Urodynamic studies were performed before transplantation in 6/7 patients. In 4 the investigation indicated bladders of capacity and compliance which were deemed suitable for transplantation. Two patients had poorly compliant bladders; one of these underwent bladder augmentation before engraftment and the other proceeded to transplantation without bladder surgery. Six patients have functioning renal allografts with a mean follow-up of 1.3 years and a mean plasma creatinine of 51.6 mumol/l. Mean glomerular filtration rate (ml/min/1.73 m2 SA) 6 months after transplantation was 76.8 and at 1 year it was 84.5. In one patient early rejection was followed by transplant nephrectomy. Careful pre-operative evaluation is mandatory for a successful outcome of renal transplantation in young boys with posterior urethral valves. PMID- 8220998 TI - Antenatal diagnosis of posterior urethral valves. AB - The antenatal histories of 42 patients with posterior urethral valves diagnosed between June 1987 and September 1990 were reviewed. The mothers of all patients had at least one ultrasound scan during pregnancy. Despite this, fetal uropathy was diagnosed in only 19 cases. The remaining 23 undiagnosed children presented acutely, all within the first 6 months of life. In 33 of 36 pregnancies scanned before 24 weeks' gestation, fetal urological pathology was undetected. Mean plasma creatinine (pCr) at presentation in the group antenatally diagnosed was 139 mumol/l and in those presenting acutely was 238 mumol/l. All pCr analysed were taken after at least 48 h of life. Renal function as measured by follow-up pCr was better in the antenatally diagnosed group during the first year of life. It would appear that a routine second ultrasound scan at 26 weeks' gestation or later would reveal more cases of posterior urethral valves and this information may improve the outcome in terms of renal function. PMID- 8220999 TI - Internal ureteric stenting following pyeloplasty reduces length of hospital stay in children. AB - The advent of totally internal ureteric stents has the potential to reduce hospital stay in paediatric pyeloplasty. Traditionally, discharge from hospital has followed removal of an external trans-anastomotic drain, usually 5 days to a week post-operatively. The use of totally internal catheters negates the need for nursing supervision by removing external attachments that a paediatric patient might inadvertently dislodge. The length of hospital stay for pyeloplasty using a Double-J ureteric stent was compared with our previous method of trans anastomotic feeding tube nephrostomy drainage. The results show the use of Double J stenting to be advantageous in paediatric pyeloplasty by decreasing considerably the length of hospital stay. This is clearly of benefit to the patient, who returns to the family setting much earlier. Despite the need for an outpatient endoscopic procedure to remove the stent, savings in treatment costs and improved efficiency of bed use are also achieved. PMID- 8221000 TI - Intravesical ureteric plication and reimplantation for megaureters in children. AB - Historically, megaureters have always been a surgical dilemma for paediatric urologists. However, the evolution of modern diagnostic and surgical methods such as tailoring, folding and plication have made it possible to ensure successful correction in most patients. We report 17 megaureters (11 refluxing and 6 obstructing) in 11 children who were treated with intravesical plication and the trans-trigonal advancement technique between January 1986 and April 1991. Results were excellent in 11 ureters and satisfactory in 4. In one ureter additional surgery was needed because of obstruction at the implantation site and in another ureter reflux persisted. Intravesical plication and reimplantation is a good alternative procedure for grossly dilated ureters owing to its low morbidity and high success rate. PMID- 8221001 TI - Inflammatory pseudotumour of the urinary bladder. PMID- 8221002 TI - Pure primary small cell carcinoma of the prostate. PMID- 8221003 TI - Endometrial stromal sarcoma invading the bladder. PMID- 8221004 TI - Bladder hamartoma. PMID- 8221005 TI - Renal transplantation in a patient with multifocal transitional cell carcinoma. PMID- 8221006 TI - Bilateral testicular teratoma in Klinefelter's syndrome. PMID- 8221007 TI - Bilateral pyelosinus extravasation secondary to probable radiation-induced ureteric fibrosis. PMID- 8221008 TI - Metastatic appendiceal carcinoma masquerading as spermatic cord tumour. PMID- 8221009 TI - Removal of ureteric stents in women without cystoscope. PMID- 8221010 TI - Total fibrotic obliteration of urethral stent. PMID- 8221011 TI - Simple technique for vasography. PMID- 8221012 TI - A sliding urethral catheter for cystometric studies. PMID- 8221013 TI - Reduction of post-operative urethral strictures by double sheath continuous flow resectoscope. PMID- 8221014 TI - Re: Rome pouch: pre-peritoneal continent ileal reservoir with hydraulic valve. PMID- 8221015 TI - Re: Suprapubic catheterisation and bowel injury. PMID- 8221016 TI - Re: The mobile lithotriptor: an answer for the smaller centre. PMID- 8221017 TI - An introduction to free radical biochemistry. AB - Free radicals are chemical species possessing an unpaired electron that can be considered as fragments of molecules and which are generally very reactive. They are produced continuously in cells either as accidental by-products of metabolism or deliberately during, for example, phagocytosis. The most important reactants in free radical biochemistry in aerobic cells are oxygen and its radical derivatives (superoxide and hydroxyl radical), hydrogen peroxide and transition metals. Cells have developed a comprehensive array of antioxidant defences to prevent free radical formation or limit their damaging effects. These include enzymes to decompose peroxides, proteins to sequester transition metals and a range of compounds to 'scavenge' free radicals. Reactive free radicals formed within cells can oxidise biomolecules and lead to cell death and tissue injury. Establishing the involvement of free radicals in the pathogenesis of a disease is extremely difficult due to the short lifetimes of these species. PMID- 8221018 TI - Measuring free radical reactions in vivo. AB - The increasing interest in the role of free radicals in the pathogenesis of human disease has led to an increased need for techniques to measure free radicals and their reactions in vivo and, most importantly, in the clinical situation. Free radicals are extremely reactive and thus short lived. Consequently, free radicals are not amenable to direct assay and free radical activity is usually assessed by indirect methods such as measurement of the various end products of reactions with lipids, proteins and DNA. A vast array of analytical techniques has been developed to measure these end products though not all of them are applicable to the clinical situation where the only samples normally available are blood, urine and expired breath. Lipid peroxidation is the most intensively studied process and provides a number of possibilities for assays. Protein and nucleic acid oxidation are attracting increasing attention at the present time. The techniques currently available, however, are limited to semi-quantitative assays of damage to broad classes of biomolecules and there is an urgent need for more specific and informative methods. PMID- 8221019 TI - Free radicals in inflammation: second messengers and mediators of tissue destruction. AB - In recent years it has become increasingly apparent that, in man, free radicals play a role in a variety of normal regulatory systems, the deregulation of which may play an important role in inflammation. As examples, we discuss the second messenger roles of: NO in the regulation of vascular tone, O2.- in fibroblast proliferation and H2O2 in the activation of transcription factors such as NF kappa B. Other control mechanisms, the physiological function of which may be perturbed in inflammation, include: the oxidative modification of low density lipoprotein, the oxidative inactivation of alpha-1-protease inhibitor, DNA damage/repair and heat shock protein synthesis. At sites of inflammation, increased free radical activity is associated with the activation of the neutrophil NADPH oxidase and/or the uncoupling of a variety of redox systems, including endothelial cell xanthine dehydrogenase. Although free radicals, thus produced, have the capacity to mediate tissue destruction, either alone or in concert with proteases, we argue that disturbances in the second messenger and regulatory activities of free radicals may also contribute significantly to the inflammatory process. PMID- 8221020 TI - Oxidative mechanisms in carcinogenesis. AB - Cancer in humans and animals is a multistep disease process. In this process, a single cell can develop from an otherwise normal tissue into a malignancy that can eventually destroy the organism. The complex series of cellular and molecular changes that occur through the development of cancers can be mediated by a diversity of endogenous and environmental stimuli. Active oxygen species and other free radicals have long been known to be mutagenic; further, these agents have more recently emerged as mediators of the other phenotypic and genotypic changes that lead from mutation to neoplasia. Free radical production is ubiquitous in all respiring organisms, and is enhanced in many disease states, by carcinogen exposure, and under conditions of stress. Free radicals may therefore contribute widely to cancer development in humans. This review explores the molecular mechanisms through which free radicals can participate in the carcinogenic process. PMID- 8221021 TI - Free radicals and myocardial reperfusion injury. AB - Ischaemic myocardial tissue will, inevitably, necrose if blood flow is not restored. Whilst reperfusion is always beneficial in terms of potential recovery of heart muscle, reperfusion in itself is believed to bring about cellular injury. While the causes of this 'reperfusion injury' are apparently multifactorial, there is now an increasing body of evidence to suggest that oxygen free radicals play a major role in the pathogenesis of reperfusion injury. The initial evidence for this hypothesis was indirect, based on the ability of free radical scavengers to limit myocardial injury in animal models. More recent work has utilised the highly specific technique of electron spin resonance (ESR) spectroscopy and ESR spin trapping to detect the free radical species. The evidence for free radical production on myocardial reperfusion will be presented along with details of human studies. The potential for a therapeutic intervention will also be briefly discussed. PMID- 8221022 TI - Involvement of oxygen radicals in shock related cell injury. AB - Shock-related organ failure evolves from a variety of starting points--ischemia, reperfusion, non-bacterial or bacterial inflammation--several mechanisms are involved. In addition to the effects of xanthine oxidase after ischemia/reperfusion, toxic oxygen species from phagocytes that accumulate in both intra- and extravascular tissue spaces are of central importance. A critical event is the contact (adhesion) of leukocytes to endothelial cells, which consequently are the targets for leukocyte products. Damage of membranes by lipid peroxidation and by exposure to mediators such as platelet activating factor (PAF), leukotrienes and proteases, leads to increased permeability, tissue oedema and organ dysfunction. Thus antioxidants and other agents that control phagocyte function are likely to contribute to the protection of the permeability barrier in shock states. PMID- 8221023 TI - Lipid peroxidation and its role in atherosclerosis. AB - A crucial step in the pathogenesis of atherosclerosis is believed to be the oxidative modification of low density lipoprotein (LDL). The oxidation of LDL is a free radical driven lipid peroxidation process and the aldehyde products of lipid hydroperoxide breakdown are responsible for the modification of the LDL apoprotein. Aldehyde-modified apoB protein has altered receptor affinity, causing it to be scavenged by macrophages in an uncontrolled manner with the development of foam cells and the initiation of the atherosclerotic lesion. The aldehydic products of lipid peroxidation may also be involved in other aspects of the development of the lesion. The oxidation of LDL may be prevented by its endogenous antioxidant compounds, most prominent of which is alpha-tocopherol. Consequently, an improved antioxidant status may offer possibilities for the prevention of this major disease. PMID- 8221024 TI - Free radicals in brain metabolism and pathology. AB - Reactive oxygen metabolites (ROM), namely superoxide and hydroxyl free radicals and hydrogen peroxide, are produced as a consequence of the physiological metabolic reactions and functioning of the central nervous system. ROM have also been implicated in the aetiopathogenic processes of a number of pathological conditions of the brain. While primarily indirect, evidence for this view is accumulating, and credence for the participation of free radical oxidative interactions in promoting tissue injury in such conditions as brain trauma, ischaemia, and toxicity, and in neurodegenerative diseases such as Parkinson's disease, Alzheimer's dementia, multiple sclerosis, and lipofuscinosis, is growing. Concomitant with this new understanding of the injurious role of free radical oxidants in neural pathology, is the increasing appreciation for the need for both fundamental and clinical research into the development of the potential preventative and therapeutic benefits that are now being foreseen for a variety of antioxidant nutritional and pharmacological interventions. PMID- 8221025 TI - Free radicals and lung disease. AB - The involvement of reactive oxygen species (ROS) in the pathogenesis of several lung diseases/injuries has been suggested. ROS are believed primarily to be generated by leukocytes (e.g. infiltrating neutrophils) although other ROS generating systems such as the xanthine/xanthine oxidase system may also be of importance. ROS may through oxidative changes exert a number of toxic effects which have been demonstrated in many different biological systems. At limited oxidative stress events such as modification of receptor activity and signalling, as well as release of endogenous mediators of inflammation may occur. One such ROS induced event, probably of importance for several lung diseases, is arachidonic acid (AA) release and metabolism to active product(s). In the lung, the release of AA results in both vaso- and bronchoconstriction, primarily caused by thromboxane A2. The molecular events leading to oxidant induced AA release and thromboxane formation are only partially elucidated. PMID- 8221026 TI - Liver damage due to free radicals. AB - The involvement of free radical reactions in the pathogenesis of liver injury has been investigated for many years in a few defined experimental systems using carbon tetrachloride, excess iron or ethanol as prooxidant agents. More recently, the hepatotoxicity of several other free radical-generating compounds has been characterised mainly in the rat hepatocyte model. In particular, the mechanisms by which drugs like paracetamol, halothane, paraquat or conditions such as ischemia-reperfusion exert their damaging activity to the liver have mostly been clarified. Since we are not trying to cure diseases occurring only in rats, the likely relevance of free radical reactions also in the genesis and progression of human liver injury has been carefully considered. Increasing evidence of free radical involvement is reported for chronic ethanol intoxication and iron overload, but the most striking proof of a causative role of ree radical chain reactions, namely lipid peroxidation, in the acute lethal damage of the hepatocyte has been obtained so far in ischemic hepatitis. PMID- 8221027 TI - Involvement of reactive oxygen species in kidney damage. AB - There is considerable evidence suggesting that reactive oxygen species (ROS) are implicated in the pathogenesis of ischemic, toxic, and immunologically-mediated renal injury. In experimental renal ischemia, ROS sources include the electron transport chain, oxidant enzymes (xanthine oxidase), phagocytes, and auto oxidation of epinephrine. ROS cause lipid peroxidation of cell and organelle membranes and, hence, disruption of the structural integrity and capacity for cell transport and energy production, especially in the proximal tubule segment. In experimental immune glomerulonephritis, ROS are generated by both infiltrating blood-borne cells (polymorphonuclear leukocytes and monocytes) and resident glomerular cells, mainly mesangial cells. Their formation results in morphologic lesions and in modifications of glomerular permeability to proteins through activation of proteases and reduction of proteoglycan synthesis. Additionally, they promote a reduction in glomerular blood flow and glomerular filtration rate through liberation of vasoconstrictory bioactive lipids (prostaglandins, thromboxane, and platelet activating factor) and, possibly, inactivation of relaxing nitric oxide. Further studies are needed to address the role of ROS in human glomerular diseases. PMID- 8221028 TI - Free radicals and muscle damage. AB - Muscle tissue is unique in its requirement and ability to undertake very rapid and co-ordinated changes in energy supply and oxygen flux during contraction. Several studies have suggested that this renders the tissue particularly prone to oxygen radical-mediated damage. Free radicals have been postulated to play a role in muscle damage induced by different forms of exercise and in various pathological disorders, such as the muscular dystrophies, malignant hyperthermia and alcoholic myopathy. However, conclusive evidence for a fundamental role for free radicals and protective effect of antioxidants remains elusive in all these situations and much further work on the relevant pathogenetic mechanisms is still required. PMID- 8221029 TI - Diabetes mellitus and free radicals. Free radicals, transition metals and oxidative stress in the aetiology of diabetes mellitus and complications. AB - Diabetes mellitus is a syndrome initially characterized by a loss of glucose homeostasis. The disease is progressive and is associated with high risk of atherosclerosis, kidney and nerve damage as well as blindness. Abnormalities in the regulation of peroxide and transition metal metabolism are postulated to result in establishment of the disease as well as its longer term complications. Diabetes mellitus is associated with oxidative reactions, particularly those which are catalyzed by decompartmentalized transition metals, but their causative significance in diabetic tissue damage remains to be established. PMID- 8221030 TI - Involvement of free radicals in ageing: a consequence or cause of senescence. AB - Free oxygen radicals are increasingly discussed as important factors involved in the phenomenon of biological ageing. Higher formation rates of free radicals from senescent animals observed in isolated biological materials (mainly in mitochondria), accumulation of free radical damage and changes of antioxidant capacities appear to prove the correctness of this assumption. In the present review these findings are critically examined in order to evaluate whether free radicals do contribute to the initiation and/or propagation of ageing. It is concluded that data available so far do not allow a definite answer to this question although, free radicals are very likely to contribute considerably to the development of stochastic disorders observed during the progress of ageing. PMID- 8221031 TI - Free radical disorders of preterm infants. AB - In recent years increasing experimental and clinical data have provided compelling evidence for the involvement of oxygen free radicals in the 3 main disorders of prematurity--chronic lung disease, retinopathy of prematurity and intraventricular haemorrhage. Infants born prior to 30 weeks gestation or weighing less than 1500 g at birth appear to be most at risk. They are very underdeveloped and as a consequence of the immaturity of their lungs often require intense respiratory support, including the provision of supplemental oxygen. The theoretical basis for free radical involvement in these disorders is that oxygen centred radicals and related reactive oxygen metabolites are formed too rapidly to be detoxified by the antioxidant defence mechanisms in specific tissues. In the case of chronic lung disease, the evidence currently favours excess oxygen (hyperoxia) as the cause of the greater oxygen free radical production, whereas in retinopathy of prematurity and intraventricular haemorrhage, it is proposed that low oxygen tensions (hypoxia) followed by periods of reoxygenation is the more likely stimulus for excess radical formation. PMID- 8221032 TI - Prospects for the prevention of free radical disease, regarding cancer and cardiovascular disease. AB - Free radicals may be involved in the aetiology of cancer and cardiovascular diseases. In epidemiological studies poor plasma levels of all essential antioxidants are associated with increased relative risks; in particular, low levels of carotene and vitamin E with the risk of cancer and ischemic heart disease, respectively. The studies suggest that for optimal synergistic protection the plasma antioxidant levels should simultaneously exceed the threshold values of 28-30 mumol/l lipid-standardized vitamin E, 40-50 mumol/l vitamin C, 0.4-0.5 mumol/l carotene and 2.2-2.8 mumol/l lipid-standardized vitamin A. However the preventive efficacy of an optional antioxidant status is still to be proven in randomized intervention trials. Although these antioxidant micronutrients may be the primary protective components of vegetable-rich 'preventive' diets, the potentials of other plant components await exploration, eg carotenoids other than beta-carotene, bioflavonoids and oxygen-sensitive B vitamins. PMID- 8221033 TI - Prospects for treatment of free radical-mediated tissue injury. AB - Toxic metabolites of oxygen are generated normally by aerobic metabolism in cells and this generation can significantly increase in certain pathologic conditions. When endogenous antioxidant defense capabilities are exceeded by this oxidant flux, tissue injury occurs. This process can be intercepted pharmacologically at different levels with agents that scavenge reactive oxygen metabolites, block their generation, or enhance endogenous antioxidant capabilities. In many situations, such as the treatment of post-ischaemic reperfusion injury, efficacy of antioxidant is related primarily to the proportionate magnitude of the total injury sustained that is due to an antioxidant mechanism. This approach has shown great promise in animal models of clinical problems and has been tested successfully in early, controlled clinical trials. PMID- 8221034 TI - Equine viral arteritis--a challenge for the British horse industry. PMID- 8221035 TI - Horserace Betting Levy Board recommendation for the 1993 covering season. Common code of practice for equine viral arteritis. PMID- 8221036 TI - George Fleming Prize. PMID- 8221037 TI - The Royal (Dick) School of Veterinary Studies: the past, the present and the future. PMID- 8221038 TI - Secreted regulatory proteins: new opportunities for immunological research in domesticated species. PMID- 8221039 TI - The life and work of William Dick. PMID- 8221040 TI - Neutrophil chemotaxis in the horse is not mediated by a complex of equine neutrophil elastase and equine alpha-1-proteinase inhibitor. AB - Studies have demonstrated that as a result of proteolytic inactivation or complex formation (with neutrophil elastase), human alpha-1-proteinase inhibitor (API) becomes a potent chemoattractant for human neutrophils. The present study aimed to investigate the in vitro chemotactic response of equine neutrophils to an equivalent complex of equine API and neutrophil elastase. No evidence of neutrophil migration was observed towards purified complex derived from equine neutrophil elastase and the Spi 1 isoform of equine API, or to crude mixtures of porcine pancreatic elastase and unseparated equine API isoforms, although the same neutrophil preparations actively migrated towards zymosan activated plasma. It was concluded that, in the horse, complexes of API are not involved in the migration of neutrophils to sites of inflammation. PMID- 8221041 TI - The dangers of disease transmission by artificial insemination and embryo transfer. AB - This review summarizes the major infectious diseases of the three major agricultural species (cattle, sheep and pigs) and horses, and presents the evidence for and against the possibility of infectious agents being transmitted between animals via the venereal route or by the use of semen or early embryos in commercial artificial insemination (AI) or embryo transfer (ET). Cattle feature most prominently in the widespread distribution of frozen semen, and national and international organizations have set out guidelines to work towards disease-free bull studs with semen free from potential pathogens. With the control of major epizootic diseases, attention has been focused on such diseases as IBR, BVD and blue tongue, where clinical signs are rarely evident but the detection of virus in semen is of great importance. New information on the relevance of bacterial disease such as Mycobacterium paratuberculosis, campylobacteriosis and leptospirosis is reviewed, along with details of the mycoplasma and ureaplasma species of the bull's genital tract. Bovine spongiform encephalopathy (BSE) has attracted much research and semen is not regarded as a source of infection. New work on the pathogenesis of a number of diseases and the use of new biotechnology in diagnosis is included. The International Embryo Transfer Society (IETS) has encouraged a great deal of experimental work--much originating in Canada--on the risk of transmission of disease from donors to recipients via a 7-day-old blastocyst. There has been much success in demonstrating that with an approved protocol of handling the embryos, to date there is very little danger in disease transmission with both viruses and bacteria. The mycoplasma group appear more intractable and the role of BSE is still being evaluated. In sheep, scrapie, Brucella ovis infection and blue tongue feature in current work. In the pig there is a surge in international movement of pig semen, and Aujeszky's disease and the new so-called Blue Ear disease feature prominently. Much work is in progress on infectious agents likely to be found in the semen of stallions, with an expanding trade in the international movement of chilled and frozen semen. Equine embryo transfer experiments are hampered by the very limited number of embryos available. Reference is also made to the further risk of disease transmission by in vitro manipulated embryos. PMID- 8221042 TI - Current research on ovine cytokines. AB - Cytokines are key mediators of the immune system, dictating the quality of the host response to infection. The importance of such immune mediators to the development of immune and inflammatory responses has emerged from work in mouse and man, however it has now become necessary to produce the equivalent (and novel) cytokines in ruminants. Over the past three years recombinant DNA techniques have allowed the cloning of numerous ovine cytokines. These include interleukins -1, -2 and -3 (IL-1, -2 and -3), interferon-gamma (IFN-gamma), ovine trophoblast protein (oTP-1), tumour necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF). The predicted amino acid sequences of these ovine proteins show varying degrees of similarity with the equivalent human proteins thus explaining why some of the cytokines are not biologically cross-reactive between species. Recombinant ovine proteins have been produced for IFN-gamma, oTP-1, IL-1, IL-3 and GM-CSF. Their biological activities are very similar to those of their human counterparts. Although it is too early to tell whether the recombinant ovine proteins will be of use in the treatment or prophylaxis of infectious disease, work in cattle and pigs has indicated the potential usefulness of cytokines in this role. PMID- 8221043 TI - Sequence of an exon of the feline p53 gene--mutation in a lymphosarcoma. AB - Exon 8 of tumour suppressor gene p53 was sequenced in domestic cats and showed remarkable similarity to the human sequence. Only four of the 13 nucleotide differences gave rise to interspecific amino acid differences. In an investigated lymphosarcoma we detected a mutation cgg --> tgg (arginine --> tryptophan) in codon no. 282. PMID- 8221044 TI - The depletion of glycogen stores and indices of dehydration in transported broilers. AB - Broilers were either not transported or were transported for 2, 4 or 6 hours after having been subjected to food withdrawal times of less than one hour or of ten hours. The birds were then slaughtered using normal commercial practices. The longer period of food deprivation reduced liver weight and glycogen content, and circulating glucose concentrations. It also elevated the ultimate pH value (pHu) in the biceps muscle and by implication, therefore, reduced its glycogen content. With longer journey times, liver weight and glycogen content decreased. Transport had an inconsistent effect on glycogen concentration in the pectoral muscle but progressively reduced its pHu. In contrast, pHu in the biceps progressively increased, by implication because transport depleted muscle glycogen levels. Birds transported further had higher concentrations of total protein in their plasma which, though this was not significant, also had a higher osmolality. This suggests that transported birds became dehydrated. Additionally, the depletion of body glycogen stores might be associated with the perception of fatigue. PMID- 8221045 TI - Silent cerebral embolism caused by neurologically symptomatic high-grade carotid stenosis. Event rates before and after carotid endarterectomy. AB - Fourteen symptomatic patients with severe extracranial internal carotid artery stenosis (> or = 70% of luminal narrowing) were monitored using long-term transcranial Doppler ultrasonography to determine the rate of clinically silent embolism of the ipsilateral middle cerebral artery. Before carotid endarterectomy (all patients being treated with intravenous heparin), 462 such events occurred during a total monitoring time of 45 h. Statistical analyses of the inter-event intervals and of the relationship between events and cardiac cycle revealed random occurrence. While the ipsilateral events were found in each subject, silent embolism of the contralateral middle cerebral artery occurred in only four patients each of whom had angiographically proven intracranial cross-flow from the symptomatic carotid territory (39 embolic events during 7 h monitoring time). The other 10 patients showed no contralateral embolism (10 h monitoring time). Five or more days (median 10 days) after surgery and cessation of intravenous anticoagulation the ipsilateral event rate had dropped to 13 in 33 h (P < 0.001) and the contralateral rate to zero. This suggests that the stenosed arterial segment is the main source of cerebral embolism detectable with ultrasound in symptomatic patients with high-grade internal carotid stenosis, and that carotid endarterectomy substantially reduces the rate of these events. Since reduction of ipsilateral stroke risk by successful endarterectomy is known to be of a similar degree in patients as those studied here, transcranially detected embolism may represent a new marker of disease activity of extracranial carotid artery stenosis. PMID- 8221046 TI - Examination of distal involvement in cisplatin-induced neuropathy in man. An electrophysiological and histological study with particular reference to touch receptor function. AB - Cisplatin is a widely used anti-neoplastic agent with dose-dependent sensory neuropathy as a major side-effect. The mechanism for the neuropathy is poorly understood; it may be caused by a lesion of the dorsal root ganglion cells or by a distal axonopathy. This distinction is important since regeneration in a neuronopathy is impossible, whereas recovery may occur if the axon is affected only distally. The most distal part of the sensory nerve fibre is, however, not accessible for conventional electrophysiological examination. To ascertain whether the distal receptor-associated part of the fibre is involved, we have used a method previously untested in patients with neuropathy. In 26 males treated with cisplatin for testicular cancer 3-6.5 years previously, and in 22 normal males, the compound sensory action potentials evoked by a tactile probe were recorded through needle electrodes placed close to the sural and median nerves. The responses were compared with action potentials evoked by electrical stimulation of the same nerves. Biopsies from the distal sural nerve at the dorsolateral aspect of the foot were obtained in three patients and in four subjects not treated with cisplatin. Sixteen patients had received a conventional dose (307-435 mg/m2) of cisplatin and 10 patients had received a high dose (553 1197 mg/m2). Two-thirds of the conventional dose patients and all the high dose patients had mild to severe sensory loss and reduced or absent tendon reflexes. The amplitude of the electrically evoked sensory action potential decreased with increasing dose of cisplatin and was correlated with the reduction of vibration sense. Tactile responses, probably originating mainly from Pacinian corpuscles, were, with the exception of two high dose patients, recorded from all sural and median nerves. The two high dose patients without a tactile response had a severely reduced or no electrically evoked response at the sural nerve. The sural nerve biopsies from high dose patients showed loss of large fibres; Pacinian corpuscles were obtained in two of these patients and contained normal axons. Our findings do not suggest that cisplatin causes a primarily distal lesion with sparing of more proximal parts of the peripheral nerve. We interpret the results as being consistent with a neuronopathy affecting primarily large sensory neurons. Brainstem and somatosensory evoked potentials and H-reflexes suggested that the spinal cord and brainstem were affected as well. PMID- 8221047 TI - The neurotrophic analogue of ACTH(4-9), Org 2766, protects against experimental allergic neuritis. AB - Demyelinating diseases such as multiple sclerosis or the Guillain-Barre syndrome originate from an autoimmune response resulting in the degradation of myelin and impaired neuronal function. Prophylactic administration of the neurotrophic peptide, H-Met(O2)-Glu-His-Phe-D-Lys-Phe-OH [an ACTH(4-9) analogue], to rats with experimental allergic neuritis, a model for the Guillain-Barre syndrome, markedly suppresses the clinical symptoms, protects against loss of motor coordination and prevents the degeneration of myelinated axons in the affected peripheral nerve. Therefore, this peptide may provide a new approach to the therapy of peripheral demyelinating polyneuropathies. PMID- 8221048 TI - Gadolinium enhancement increases the sensitivity of MRI in detecting disease activity in multiple sclerosis. AB - There is now widespread agreement that serial brain MRI is useful in monitoring treatments designed to modify the course of multiple sclerosis. It has been less clear whether gadolinium enhancement is needed. We therefore compared the relative sensitivity of long repetition time (TR) spin echo (SE) and gadolinium enhanced short TR SE sequences in detecting active lesions. A blind analysis of the two sequences was performed in 26 untreated patients with early relapsing- remitting (19) or secondary progressive (seven) multiple sclerosis who underwent monthly MRI on four occasions (one baseline and three follow-up). Active lesions were defined as either new or enlarged lesions on long TR SE, or new or persistent enhancing lesions on short TR SE. In one patient there were 144 active lesions, all of which were seen with enhancement on short TR SE, but only 17 were seen on long TR SE. Amongst the remaining 25 cases, a total of 106 active lesions were seen: 68 (64%) were seen only with enhancement on short TR SE, 16 (15%) were seen only on long TR SE, while 22 (21%) were active on both sequences. We conclude that gadolinium enhancement markedly increases the sensitivity of monthly brain MRI in monitoring the treatment of relapsing--remitting or secondary progressive multiple sclerosis. With this frequency of scanning, a post contrast short TR SE sequence is the most sensitive method for detecting active lesions. The smaller yet still substantial incidence of active lesions seen only on the long TR SE sequence suggests that it should also be obtained. PMID- 8221049 TI - Ocular torsion and perceived vertical in oculomotor, trochlear and abducens nerve palsies. AB - Ocular torsion (OT) and subjective visual vertical (SVV) were determined in acute and chronic oculomotor (n = 6), trochlear (n = 21) and abducens (n = 7) palsies separately for each eye in the primary position with the head upright. Ocular torsion measured by fundus photographs was not only within normal range in all abducens palsies, but unexpectedly also in 68% of third and fourth nerve palsies which involve oblique eye muscles. Pathological OT, when measurable, was slight (2 degrees - 8 degrees), monocular and occurred either in the paretic or in the nonparetic eye. Subjective visual vertical tilts were more frequent (67% of third and fourth nerve palsies) although mostly small in amplitude (1 degree - 6 degrees). They were confined either to the paretic or the nonparetic eye depending on the duration of the palsy. Determinations of SVV were always normal under binocular viewing conditions. The dissociated occurrence of OT and SVV tilts in the paretic or the nonparetic eye was dependent on the acuteness of the palsy and reflected sensory and/or motor compensation mechanisms. Third and fourth nerve palsies cause only minor and unpredictable monocular OT and SVV tilts as distinct from the frequent binocular and conjugate tilts seen in patients with acute unilateral brainstem lesions. PMID- 8221050 TI - Abnormal spatial localization with trigeminal-oculomotor synkinesis. Evidence for a proprioceptive effect. AB - We examined spatial localization, using open-loop pointing to visual targets, in a patient with a congenital trigeminal-oculomotor synkinesis. This patient demonstrated abnormal co-activation of the left medial rectus muscle when the left lateral pterygoid contracted. Because one eye could be deviated in the absence of a normal oculomotor innervational command, the efference copy (derived from monitoring of central oculomotor commands) could be dissociated from the proprioceptive afferent signal (determined by the mechanical state of the extraocular muscles). Under conditions of monocular viewing with the normal right eye, when the covered left eye was adducted by the aberrant trigeminal innervation, the patient pointed to the left of the actual position of the target. This finding indicates that proprioceptive afference from the adducted, covered left eye was used in the process of spatial localization. While synkinetic adduction produced a shift in pointing in the opposite direction of rotation of the non-viewing eye, previous studies using passive deviation of the non-viewing eye in normal subjects reported a shift in pointing in the same direction as eye rotation (Gauthier et al., 1990; Bridgeman and Stark, 1991). We propose that this discrepancy is due to the different effects of passive eye rotation and active muscle contraction on the tendon organs of the extraocular muscles. On this basis, we hypothesize that the tendon organs, rather than the muscle spindles, are primarily responsible for the transduction of proprioceptive information about eye position in the orbit. PMID- 8221051 TI - Study of selective reaching and grasping in a patient with unilateral parietal lesion. Dissociated effects of residual spatial neglect. AB - In the present study we investigated the possibility of a dissociation between the visual control of reaching and the visual control of grasping in a prehension task. To this purpose we studied the kinematics of prehension movements in a patient with a right parietal lesion and in six right-handed healthy control subjects. The task we used was one in which the subjects had to reach and grasp target objects in the presence or absence of a simultaneously presented distractor object. All stimuli were presented in the space ipsilateral to the lesion. The distractor could be either of the same or different size to the target object and was presented either to the right or to the left of the target. The following parameters of the prehension 'transport' component were analysed: wrist trajectory, transport time, tangential peak velocity, acceleration. Maximal finger aperture, time to maximal finger aperture, peak acceleration and time to peak acceleration of grip aperture were the parameters of the 'grasping' component analysed. The results showed that, although the patient had no misreaching, her hand trajectory deviated abnormally towards the distractor position when the distractor was to the right (ipsilateral) side of the target. In contrast, the grasp kinematics was not affected by the distractors, even when the size of the right distractor was different from the target. It appears, therefore, that the attentional shift towards the ipsilesional side, typical of neglect patients, determines a surprising dissociation in motor control. In the presence of a right distractor, the patient plans and partially executes a reaching movement towards that object and simultaneously performs a grasping movement towards a second object, i.e. the centrally located target. The presentation of distractors had no effects on the prehension kinematics of the control subjects. PMID- 8221052 TI - Disturbed frontal regulation of attention in Parkinson's disease. AB - Parkinson's disease is characterized not only by tremor, akinesia and rigidity, but also by frontal cognitive dysfunction, that can be understood as a disturbance in the 'Supervisory Attentional System' (SAS). This concept refers to a system, located in the frontal cortex, that regulates attentional processes under novel, non-routine conditions. The hypothesis that cognitive dysfunction in Parkinson's disease results from a disturbance in the SAS was investigated by recording 'processing negativity' in 33 parkinsonian patients and 17 controls. Processing negativity is an event-related potential that reflects neuronal activity during selective attention. The contribution of the frontal cortex to selective attention can be studied directly using processing negativity. Parkinsonian patients were also scored for clinical symptoms and subjected to a neuropsychological test battery. Processing negativity was clearly disturbed in the parkinsonian patients. Moreover, parkinsonian patients with the lowest scores on 'frontal' neuropsychological tests such as Stroop, Trail Making and Word Fluency, also had the lowest processing negativity. Our results support the hypothesis that cognitive dysfunction in Parkinson's disease might be understood as a disturbance in the frontal regulation of attentional processes. Degeneration of the dopaminergic mesocortical innervation of the frontal cortex in Parkinson's disease is a possible neurochemical substrate of these frontal attentional disturbances. PMID- 8221053 TI - Contrasting mechanisms of impaired attentional set-shifting in patients with frontal lobe damage or Parkinson's disease. AB - Tests which assess the ability to shift cognitive set modelled after the Wisconsin Card Sorting Test are particularly sensitive to impairments in patients with Parkinson's disease as well as in patients with frontal lobe damage. However, the underlying mechanisms responsible for the similar deficits observed in the two patient groups are not well understood and may not be identical. For example, an apparent deficit in set-shifting ability may reflect either an impairment in the ability to shift from a perceptual dimension which has previously commanded attention (i.e. 'perseveration'), or in the ability to shift to an alternative perceptual dimension which has previously been irrelevant (i.e. 'learned irrelevance'). In this study, the performance of both medicated and non medicated patients with Parkinson's disease were compared with a group of neurosurgical patients with localized excisions of the frontal lobes on a novel task designed to assess the relative contribution of 'perseveration' and 'learned irrelevance' to impaired set-shifting ability. Patients with frontal lobe damage were worse than controls in their ability to shift attention from a previously relevant stimulus dimension. Medicated patients with Parkinson's disease were worse at shifting to a previously irrelevant dimension. In contrast to both groups, nonmedicated patients with Parkinson's disease were impaired in both conditions. These results suggest that the gross set-shifting deficits reported in both frontal lobe patients and patients with Parkinson's disease may involve fundamentally different, though related, cognitive processes, and that these may be differentially affected by medication. Specifically, L-dopa therapy may protect Parkinson's disease patients from preservation of attention to a formerly relevant stimulus dimension. PMID- 8221054 TI - Vestibular induced postural responses in Parkinson's disease. AB - We have tested the hypothesis that dysfunction of the vestibular control of posture is a principal cause of parkinsonian instability by measuring the body sway response induced by galvanic vestibular stimulation (0.5 mA for 2 s) in a group of patients with Parkinson's disease (n = 15). Responses were compared with those obtained from a group of age-matched control subjects (n = 10). Subjects were stimulated (randomized polarity) whilst standing with feet together, eyes closed and maintaining one of five head yaw angles. The motion of the body and the ground reaction forces were measured in three dimensions. There was no significant difference between patients and controls in the speed or direction of the induced body sway response. When the patients were subdivided into two groups according to a clinical assessment of postural deficit, the more disabled group was found to respond with significantly greater body speed than either the control group or the mildly affected patient group. However, the baseline speed of spontaneous body sway was also greater in these patients and it was found that response speed was linearly related to baseline body sway even for the control group. There were no significant differences between any of these groups in the latency to onset, latency to peak or peak amplitude of the initial horizontal ground reaction force response to the stimulus. We conclude that vestibular dysfunction does not explain the postural deficits of patients who are mildly or moderately affected by Parkinson's disease. PMID- 8221055 TI - Striatal [18F]dopa uptake in familial idiopathic dystonia. AB - It is known that most cases of idiopathic torsion dystonia (ITD) are inherited in an autosomal dominant fashion. Despite clarification of the underlying genetic defect, no consistent structural lesion has been identified in ITD, and it is probable that a biochemical disturbance is the basis of the disorder. To determine whether there is impaired function of the nigro-striatal dopaminergic terminals in ITD we studied 11 subjects with generalized ITD and a positive family history using [18F]dopa and PET scanning. Of these 11 patients, eight had putamen [18F]dopa uptake within the lower half of the normal range, while three had uptake reduced by > 2 SDs below the normal mean. The lowest putamen [18F]dopa influx constants were found in the most disabled patients. As these reductions were mild it is unlikely that abnormalities of the nigro-striatal dopaminergic pathway are the primary determinant of either the nature of the severity of dystonic symptoms. In addition, we studied three presumed carriers of the ITD gene. These subjects all had normal striatal [18F]dopa influx constants suggesting that [18F]dopa PET is unsuitable as a screening tool for ITD. PMID- 8221056 TI - Patterns of central motor reorganization in hemiplegic cerebral palsy. AB - Central motor reorganization was studied in 33 subjects with hemiplegic cerebral palsy. Corticospinal projections were investigated using focal magnetic stimulation of the motor cortex. Reflex pathways were examined with digital nerve stimulation. Cross-correlation analysis of multi-unit EMG was used to detect activity in branched common stem last order presynaptic inputs to motor neuron pools. The neurophysiological findings were related to the clinical outcome. In 21 of the subjects studied (64%), there was evidence for reorganization of central motor pathways. The clinical and neurophysiological findings revealed two different forms of reorganization. In both forms focal magnetic stimulation demonstrated novel ipsilateral motor pathways from the undamaged motor cortex to the hemiplegic hand. Ipsilateral projections were not demonstrated from the damaged motor cortex. Eleven subjects had intense mirror movements. In these subjects cross-correlation analysis and reflex testing suggested that corticospinal axons had branched abnormally and projected bilaterally to homologous motor neuron pools on both sides of the spinal cord. The remaining 10 subjects did not have intense mirror movements and in these subjects there was no evidence for last order branching of corticospinal axons. It was found that good function of the hemiplegic hand was associated with the presence of EMG responses in that hand following magnetic stimulation of the contralateral motor cortex. When EMG responses were absent, hand function was poor unless the subject had intense mirror movements. PMID- 8221057 TI - The human dorsal hippocampal commissure. An anatomically identifiable and functional pathway. AB - The hippocampal commissural system in the human brain was found to be similar to that of non-human primates. Three normal serially sectioned human brains were studied in coronal and sagittal sections. Morphological criteria that had been validated in experimental studies on the hippocampal commissures in monkeys (Amaral et al., 1984; Demeter et al., 1985; Lamantia and Rakic, 1990) were applied to the study of these human brains. It was found that while a further reduction in the ventral hippocampal commissure has taken place in human phylogeny leading to its near or total disappearance, the dorsal hippocampal commissure is well developed and represents a sizable fibre tract. It crosses the midline under the rostral portion of the splenium and the caudal part of the body of the corpus callosum. Its fibres as they travel between the splenium and the hippocampal formation attach themselves to the fornix and the inferior portion of the forceps major of the corpus callosum. In its morphology the human dorsal hippocampal commissure fully conforms to that reported in experimental tracer studies in the monkey (Amaral et al., 1984; Demeter et al., 1985). Depth electrode EEG recordings of temporal lobe seizures show, in some instances, a pattern of contralateral spread which strongly suggests that it occurred through the dorsal hippocampal commissure. This statement is based on a careful analysis of the different patterns of contralateral spread of seizure discharge in the light of the known anatomical connections of mesial temporal structures through both direct commissural pathways and through possible indirect ones involving subcortical structures or the frontal lobe. For seizure discharges originating in mesial temporal structures of one side with spread to the contralateral hippocampus before any involvement of the contralateral isocortex the dorsal hippocampal commissure is the only likely pathway of contralateral propagation. Alternative routes appear unlikely in the light of the known primate anatomy of the commissural and other connections of the temporal lobe. Thus humans, despite claims to the contrary (Wilson et al., 1987, 1990, 1991), seem to possess a functional dorsal hippocampal commissure. Some patterns of seizure spread observed in this study which seem to utilize the dorsal hippocampal commissure may be relevant for two phenomena that are of clinical interest: (i) pure amnestic seizures (Palmini et al., 1992); (ii) false lateralization of seizure onset in extracranial EEG recordings in the type III of contralateral seizure spread as defined in this study. PMID- 8221058 TI - Amygdaloid central nucleus lesions and cholinergic blockade attenuate the response of the renal peripheral benzodiazepine receptor to stress. AB - Previous research has demonstrated that the density of peripheral benzodiazepine receptors (PBR) in rat kidney rapidly drops following exposure to 80 min of stress. The present experiments examined the contribution of the central and autonomic nervous systems in mediating this effect. Ibotenic acid lesions of the amygdaloid central nucleus (ACe), but not the lateral and basolateral amygdala, diminished the magnitude of the reduction in renal PBR binding caused by stress. Pretreating rats with methyl-scopolamine also inhibited the response of the PBR to stress. Adrenergic blockade with nadolol was ineffective. In order to test whether the PBR was under direct or indirect neural control during stress, unilateral renal denervation was performed. The stress-induced reduction in PBR binding persisted in denervated kidneys revealing that any neural control over the PBR that might exist must be indirect. Together the results suggest that the CNS may be involved in regulating the PBR during stress through the activation of intermediate, possibly hormonal, factors. The involvement of the central nervous system in the modulation of the PBR indicates the relevance of the PBR to physiological adaptations to stress. PMID- 8221059 TI - Chloride current observed as calcium-gated tail current in trigeminal root ganglion neurons of the marine catfish, Plotosus. AB - Isolated trigeminal ganglion (TRG) neurons of Plotosus in primary culture were studied with patch electrodes in a whole-cell recording configuration. When Ca currents were isolated using electrodes filled with CsCl and Ca buffer in Na- and K-free saline, a large tail current was induced. Both Ca and tail currents were blocked by 2 mM Co2+ in the bath. Reversal potential of the tail current was close to the equilibrium potential for Cl-. The reversal potential was altered by substitution of external Cl- with Br- or methanesulphonate- (MSA-). The anion permeability ratios were estimated as PBr/PCl = 1.95 and PMSA/PCl < 0.05. These results suggested that the tail current was the Ca-gated Cl current. In Plotosus neurons, two types of Ca current, high- and low-voltage activated (HVA and LVA), have been described. Both types of Ca currents were able to induce a tail current. Ba currents through both HVA and LVA channels proved to be ineffective. Upon repetitive activation, the tail current progressively increased in the presence of a nearly constant peak Ca current, and showed a more prolonged complex decay phase. When the electrode was filled with 20 mM EGTA, no tail current was induced, not even after repetitive stimulation. Therefore, the tail current seemed to be regulated by local Ca activity in the vicinity of the membrane. The Ca-gated Cl tail current may regulate the membrane excitability of TRG neurons during repetitive firing. PMID- 8221060 TI - Colocalization of D1 and D2 dopamine receptor mRNAs in striatal neurons. AB - There is evidence that D1 and D2 dopamine receptor subtypes coexist at the cellular level in the striatum and act synergistically to mediate the effects of dopamine. Other data suggest that these receptor subtypes are largely segregated in different striatal projection pathways. We used in situ hybridization in serial adjacent 4 microns sections to determine the extent of colocalization of D1 and D2 receptor mRNAs in rat striatal neurons. Cellular localization of D1 and D2 receptor mRNA was performed on section pairs that were hybridized with 35S labeled cDNA or oligonucleotide probes directed against non-homologous regions of D1 and D2 receptor mRNAs. We found that 26-27% of striatal cells containing one receptor subtype also contained the other subtype. Thus, although D1 and D2 receptors are segregated in the majority of striatal neurons, a substantial number of striatal neurons coexpress both dopamine receptor mRNA subtypes. Our findings provide anatomic support for many of the functional interactions that have been described for D1 and D2 receptors. PMID- 8221061 TI - Spinal cord motoneurons express p75NGFR and p145trkB mRNA in amyotrophic lateral sclerosis. AB - In the present study, in situ hybridization was used to examine the expression of nerve growth factor (NGF) receptor (p75NGFR), trk (p140trk) and trkB (p145trkB) mRNA in spinal cord sections from patients with amyotrophic lateral sclerosis (ALS). We report that the expression of p75NGFR and p145trkB mRNA is elevated in alpha motoneurons in ALS sections. However, p140trk mRNA was not expressed in either ALS or control sections. PMID- 8221062 TI - A modality-specific somatosensory area within the insula of the rhesus monkey. AB - Response properties of neurons in the monkey's granular insula (Ig) were examined with somatic, auditory, visual, and gustatory stimuli. Results indicate that a major portion of Ig is a somatic processing area exclusively, with units that have large and often bilateral receptive fields, consistent with the view that this area serves as a higher-order, modality-specific link in the somatosensory limbic pathway. PMID- 8221063 TI - Induction of HSP90 alpha heat shock mRNA after transient global ischemia in gerbil hippocampus. AB - Distribution of heat shock protein (HSP) 90 alpha mRNA induction after 10 min of transient global ischemia was investigated in gerbil hippocampus by in situ hybridization. A small amount of HSP90 alpha mRNA was normally present in hippocampal cells and the mRNA was further induced with a peak at 8 h after ischemia. In hippocampal CA1 cells that are vulnerable to ischemia, HSP90 alpha mRNA was continuously induced by 1 day and finally diminished at 2 days. The temporal profile of HSP90 alpha mRNA induction in hippocampal CA1 cells was similar to that of HSP70 mRNA reported previously, suggesting a cooperative role of HSP90 alpha with other HSPs after ischemia. PMID- 8221064 TI - Microinjections of subpicomolar amounts of NPY(13-36) into the nucleus tractus solitarius of the rat counteract the vasodepressor responses of NPY(1-36) and of a NPY Y1 receptor agonist. AB - Microinjections of neuropeptide Y (NPY) (1-36) and of the NPY Y1 agonist [Leu31,Pro34]NPY into the caudal dorsomedial part of the nucleus tractus solitarius (Sol) in the anaesthetized rat led to the development of dose-related vasodepressor and bradycardic responses. The threshold dose of the NPY Y2 agonist NPY(13-36) (50 fmol) significantly counteracted the vasodepressor actions of a close to ED50 dose of NPY(1-36) (2.5 pmol) and of the NPY Y1 agonist (5 pmol). These results indicate that NPY Y1 receptor activation in the Sol leads to the development of a vasodepressor response, which can be counteracted by NPY Y2 receptor activation in the Sol. The results support the existence of a Y2/Y1 receptor-receptor interaction in the Sol, via which NPY Y2 receptors may reduce transduction over NPY Y1 receptors. PMID- 8221065 TI - The caudal ventrolateral medulla is a source of tonic sympathoinhibition. AB - The caudal ventrolateral medulla (CVLM) contains neurons that are essential for the elaboration of the sympathoinhibitory effects of baroreceptor afferent stimulation. To determine if neurons in the CVLM also mediate a tonic inhibition of sympathetic nerve activity (SNA), we examined the effects of lesioning CVLM neurons in baroreceptor-denervated rats. The sustained increases in both arterial pressure (AP; 40 mmHg) and splanchnic SNA (200%) indicate that the discharge of neurons in the CVLM maintains a tonic sympathoinhibition that contributes significantly to the maintenance of normotensive levels of SNA and AP. PMID- 8221066 TI - Evidence for localization of the transgene product within axon terminals in the dentate gyrus of transgenic mice expressing human phenylethanolamine N methyltransferase. A light and electron microscopic immunocytochemical study. AB - The aim of this study was to examine whether protein products of a transgene are localized within axon terminals in transgenic mice. We have previously created transgenic mice containing a chimeric gene composed of the human dopamine beta hydroxylase gene promoter and the human phenylethanolamine N-methyltransferase (PNMT) cDNA. In the present study, we used an antiserum that detects specifically human PNMT but not mouse PNMT, and examined immunocytochemically the hippocampal formation of the transgenic mice. At a light microscopic level, immunoreactivity of human PNMT was found in fiber plexuses in the outer molecular layer of the dentate gyrus, and in cell bodies of layer 2 of the entorhinal cortex. At an electron microscopic level, in the outer molecular layer of the dentate gyrus, human PNMT immunoreactivity was observed in axon terminals that formed synapses with dendritic spines. The present study provides the evidence for localization of the transgene's protein products in axon terminals, suggesting axonal transport of the products. PMID- 8221067 TI - Membrane surface ruffling in cells that over-express Alzheimer amyloid beta/A4 C terminal peptide. AB - Deposition of beta/A4 amyloid in brain is a defining characteristic of Alzheimer disease (AD); however, the extent to which amyloid deposits may interfere with normal cellular processes is incompletely understood. We examined this issue by means of PC12 cells. After transfection with DNA coding for 97 amino acids of the beta/A4 C-terminal region of the amyloid precursor protein, beta/A4 antigen was visible at the cell membrane. We report that normal unstimulated PC12 cells exhibit ruffling activity at the cell surface when plated on a plastic substrate. Relative to control cells, however, those that over-expressed the beta/A4 C terminal peptide had significantly higher levels of ruffling activity, suggesting a structural and/or functional membrane modification. Similar cellular alterations, if present, in Alzheimer brain cells, may indicate disturbances in membrane-associated functions, including intercellular communication. PMID- 8221068 TI - Suppression of immunorejection against rat fetal dopaminergic neurons in a mouse brain by 15-deoxyspergualin. AB - In this study, the immunosuppressive effect of 15-deoxyspergualin (DSG) on the survivability of rat embryonic dopaminergic neurons grafted into the lateral ventricle of adult mice was investigated. DSG was administered daily in dose of 5 mg/kg for 2 weeks postgrafting, commencing on the day of transplantation, in the immunosuppressant-treated groups. Animals were then allowed to survive for 2 to 4 weeks after transplantation. Histological analysis revealed that most of transplanted rat fetal neural tissue was destroyed and scavenged in 2 weeks after transplantation in the non-immunosuppressed group. However, a large number of tyrosine hydroxylase (TH)-positive grafted neurons survived and grew postgrafting in the brain of the host administered with DSG even if they suffered from T lymphocyte infiltrations visualized by cytotoxic T cell immunohistochemistry. The results thus indicated that DSG is an potent immunosuppressant in neural transplantation as well as in transplantation of other organs in animals. It seems to be able to block, at least in part, the ability of mature specific cytotoxic T cells to lyse their targets. PMID- 8221069 TI - c-Fos expression by dorsal horn neurons chronically deafferented by peripheral nerve section in response to spared, somatotopically inappropriate nociceptive primary input. AB - Subcutaneous formalin injection into the hindpaw of rats induces c-Fos expression in neurons in the ipsilateral spinal cord dorsal horn. In laminae I and II of the dorsal horn at the junction of 4th and 5th segments of the lumbar spinal cord, neurons exhibiting c-Fos protein-like immunoreactivity (Fos-LI) are concentrated in the medial 3/4 that correspond to the terminal field of primary neurons innervating the sciatic nerve. Subacute tibial nerve section 24 h before formalin stimulation caused almost complete elimination of neurons with the formalin induced Fos-LI in the medial 1/2 (tibial territory) of the above sciatic territory of the dorsal horn. Following a longer survival period (chronic tibial nerve section of 21 days standing), neurons with the formalin-induced Fos-LI re appeared in the tibial territory. In addition, the number of neurons with the formalin-induced Fos-LI increased in the medial part of the peroneal territory (the lateral 1/2 of the sciatic territory). The results indicate that the activation of c-Fos expression in that part of dorsal horn that has been chronically deafferented by the tibial nerve section is taken over by the spared, but somatotopically inappropriate primary nociceptors. Furthermore, dorsal horn neurons outside but near the deafferented tibial nerve's territory exhibit hypersensitivity to c-Fos expression evoked by intact, somatotopically appropriate primary nociceptive input. PMID- 8221070 TI - Tyrosine kinase inhibitors, herbimycin A and lavendustin A, block formation of long-term potentiation in the dentate gyrus in vivo. AB - The effects of two specific tyrosine kinase inhibitors on long-term potentiation (LTP) in the dentate gyrus were investigated using anesthetized rats. I.c.v. injection of herbimycin A (1.74 nmol) or lavendustin A (1.31 nmol) before application of tetanic stimulation significantly inhibited the formation of LTP. However, these inhibitors, when administered after tetanic stimulation, did not affect the established LTP. These results suggest that protein tyrosine phosphorylation is involved in formation of LTP at the dentate gyrus in vivo. PMID- 8221071 TI - c-Fos proto-oncogene activity induced by mating in the preoptic area, hypothalamus and amygdala in the female rat: role of afferent input via the pelvic nerve. AB - In order to identify brain areas which receive afferent genitosensory input important for mating-induced prolactin release, we compared numbers of Fos immunoreactive (Fos-IR) cells in brains of intact estrous females 1 h after differential mating stimulation. Numbers of Fos-IR cells were approximately 3 fold higher in the preoptic area (POA), medial amygdala (mAMYG) and bed nucleus of the stria terminalis (BNST) when females received intromissions (I) from males than when they received mounts-without-intromission (M) or were taken directly from their home cage. In the ventrolateral portion of the ventromedial nucleus (VL-VMN), the paraventricular nucleus (PVN) of the hypothalamus and the midbrain central tegmental field (CTF) numbers of Fos-IR cells were significantly higher than home cage levels in groups of females exposed to males regardless of type of mating stimulation received. Bilateral transection of the pelvic nerve eliminated the increases in Fos-IR in POA and mAMYG which occurred in sham-transected females in these areas after intromissions from males. These data demonstrate that afferent input via the pelvic nerve activates cell groups within the POA, mAMYG and BNST and suggests that these areas may be involved in initiation of mating-induced prolactin surges. PMID- 8221072 TI - Calcium-destabilizing and neurodegenerative effects of aggregated beta-amyloid peptide are attenuated by basic FGF. AB - The mechanisms that contribute to neuronal degeneration in Alzheimer's disease (AD) are not understood. Abnormal accumulations of beta-amyloid peptide (beta AP) are thought to be involved in the neurodegenerative process, and recent studies have demonstrated neurotoxic actions of beta APs. We now report that the mechanism of beta AP-mediated neurotoxicity in hippocampal cell culture involves a destabilization of neuronal calcium homeostasis resulting in elevations in intracellular calcium levels ([Ca2+]i) that occur during exposure periods of 6 hr to several days. Both the elevations of [Ca2+]i and neurotoxicity were directly correlated with aggregation of the peptide as assessed by beta AP immunoreactivity and confocal laser scanning microscopy. Exposure of neurons to beta AP resulted in increased sensitivity to the [Ca2+]i-elevating and neurodegenerative effects of excitatory amino acids. Moreover, [Ca2+]i responses to membrane depolarization and calcium ionophore were greatly enhanced in beta AP treated neurons. Neurons in low cell density cultures were more vulnerable to beta AP toxicity than were neurons in high cell density cultures. Basic fibroblast growth factor (bFGF), but not nerve growth factor (NGF), significantly reduced both the loss of calcium homeostasis and the neuronal damage otherwise caused by beta AP. In AD, beta AP may endanger neurons by destabilizing calcium homeostasis and bFGF may protect neurons by stabilizing intracellular calcium levels. Aggregation of beta AP seems to be a major determinant of its [Ca2+]i destabilizing and neurotoxic potency. PMID- 8221073 TI - Cardiac baroreflex dynamics during the defence reaction in freely moving rats. AB - To determine the extent of baroreceptor reflex involvement in the cardiovascular changes observed during electrically induced defence reaction, the mean arterial blood pressure (MBP) and heart rate (HR) of conscious intact or sinoaortic baroreceptor denervated (SAD) rats were continuously recorded from indwelling cannulae during a 1-min period of electrical stimulation of the mesencephalic tectum. Electrical stimulation produced stimulus intensity-dependent behaviours including freezing at lower intensities and flight at higher intensities. The cardiovascular responses in intact rats were dependent on both the intensity and duration of the stimulus. A linear increase in MBP was observed with increasing stimulus intensities. However, while a slight bradycardia was observed during the freezing behaviour, a marked tachycardia occurred during flight. Simultaneous increases of MBP and HR were seen throughout the first 15 s of the flight response, after which the HR rapidly fell to baseline levels, whereas the MBP remained at a hypertensive plateau until the end of the stimulus. The baroreflex HR curve showed a parallel shift to the left during the first half of the freezing period, being fully reset 40 s after that. So, while the baroreflex gain remained unchanged, the reflex set point was lowered during the freezing stage of the defence reaction. The experiments with SAD rats corroborated the above data. The baroreceptor denervation reversed the freezing bradycardia to tachycardia. Moreover, the denervation potentiated the flight tachycardia and prevented its later reset. MBP responses of baroreceptor denervated rats did not differ from the sham-operated group. The sustained hypertension, thus, appears to be mediated by mechanisms other than the mere baroreceptor reflex deactivation. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221074 TI - Ventral tegmental microinjection of delta 9-tetrahydrocannabinol enhances ventral tegmental somatodendritic dopamine levels but not forebrain dopamine levels: evidence for local neural action by marijuana's psychoactive ingredient. AB - Basal extracellular levels of dopamine (DA) and its metabolites in both ventral tegmental area (VTA) and nucleus accumbens (Acb) were simultaneously monitored by in vivo brain microdialysis in laboratory rats. Microinjection of 12 micrograms or 24 micrograms delta 9-tetrahydrocannabinol (delta 9-THC), the psychoactive ingredient in marijuana and hashish, into the VTA produced a dose-dependent increase in somatodendritic DA levels in VTA but failed to produce any simultaneous change in extracellular DA in Acb. Direct delta 9-THC perfusion (5 x 10(-5) and 2 x 10(-4) M) into Acb via the microdialysis probes caused a significant increase in extracellular DA levels in Acb. These findings suggest that (1) delta 9-THC has a facilitating effect on somatodendritic DA efflux, and (2) the elevation of Acb DA levels seen in our previous studies with systemic administration of delta 9-THC may result from local actions of delta 9-THC at or near the DA terminal projections in Acb. PMID- 8221075 TI - Kainic acid lesions increase reafferentation of the striatum by substantia nigra grafts. AB - Effects of kainic acid lesions of the striatum on reafferentation of the striatum produced by intraventricular substantia nigra (SN) grafts were investigated. Rats with unilateral 6-hydroxydopamine lesions of the SN received intrastriatal kainic acid lesions or sham lesions, and then received fetal (E16) SN or sciatic nerve grafts in the lateral ventricle. The depth of reafferentation of the striatum by catecholaminergic neurites from SN grafts was significantly increased in rats with kainic acid-induced striatal lesions, as compared to the sham-lesioned controls. No reafferentation was seen in the control animals with sciatic nerve grafts. These data suggest that striatal injury promotes the growth of dopaminergic neurites from SN grafts. PMID- 8221076 TI - Cardiac and sympathetic effects of middle cerebral artery occlusion in the spontaneously hypertensive rat. AB - Acute increases in sympathetic activity, plasma catecholamine concentrations and myocardial damage, occur following middle cerebral artery occlusion (MCAO) in Wistar rats. Hypertension is a major risk factor for stroke. The autonomic responses to MCAO in the spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats were therefore investigated. Arterial pressure (AP), heart rate (HR), renal sympathetic nerve discharge (SND), plasma catecholamines and ECG were measured in 16 SHR and 16 WKY male urethane-anesthetized rats, which were subjected to either MCAO or sham MCAO. Cerebral infarct size did not differ between SHR and WKY rats, as shown by tetrazolium staining. Initial AP was significantly higher in SHR (96 +/- 4 mmHg) than in WKY (70 +/- 1 mmHg; P < 0.05). No significant differences in initial HR or plasma catecholamine levels were observed between SHR and WKY. By 6 hours after MCAO, AP, SND and plasma epinephrine in SHR decreased significantly, while HR showed a significant increase. SND and plasma catecholamines in the WKY showed increases that did not reach significant levels following MCAO. The QT interval of the ECG was significantly prolonged in the WKY MCAO rats, which also had a higher frequency of cardiac myocytolysis than the other groups. Unlike the increases in autonomic variables following MCAO in Wistar rats, SHR exhibit significant decreases in SND and AP, while WKY show slight, but non-significant increases. These differences in the autonomic reaction to MCAO may reflect genetic differences in the response to cerebral ischemia. PMID- 8221077 TI - Muscarinic modulation of conductances underlying the afterhyperpolarization in neurons of the rat basolateral amygdala. AB - The excitability level of pyramidal neurons in the basolateral amygdala (BLA) is greatly increased following muscarinic receptor activation, an effect associated with an increased rate of action potential firing and reduction of the afterhyperpolarization (AHP). We impaled BLA pyramidal neurons in slices of rat ventral forebrain with a single microelectrode to examine the currents underlying the AHP and spike frequency accommodation and determine their sensitivities to muscarinic modulation. In voltage-clamp, depolarizing steps were followed by biphasic outward tail currents, consisting of rapidly decaying (IFast) and slowly decaying (ISlow) current components. These corresponded temporally with the medium and slow portions of the AHP, respectively. The reversal potential for the IFast component of the AHP tail current shifted in the depolarizing direction with increases in the extracellular K+ concentration. The amplitude of IFast was reduced during perfusion of 0-Ca2+ medium or by superfusion of TEA (1-5 mM) or carbachol (10-40 microM). It is suggested that IFast was produced by the rapidly decaying Ca(2+)-activated K+ current (IC) and the muscarinic-sensitive M-current (IM). The ISlow tail current component reversed at the estimated values for EK in medium containing either normal or elevated K+ levels. This component was eliminated by perfusion of 0-Ca2+ medium or inclusion of cyclic-AMP in the recording electrode. It was not blocked by TEA (5 mM) or apamin (50-500 nM), but was reduced by carbachol in a dose-dependent manner (IC50 = 0.5 microM). Electrical stimulation of cholinergic afferent pathways to the BLA produced inhibition of ISlow, an effect which was enhanced by eserine and prevented by atropine. Loss of the ISlow component was always accompanied by similar reductions in accommodation and the slow AHP. It was concluded that this tail current component resulted from the slowly decaying Ca(2+)-activated K+ current, IAHP. Thus, the muscarinic inhibition of IAHP contributes to the enhanced excitability exhibited by BLA pyramidal neurons following cholinergic stimulation. PMID- 8221078 TI - Response of neurons in the dorsal motor nucleus of the vagus to thyrotropin releasing hormone. AB - Autonomic motoneurons in the dorsal motor nucleus of the vagus (DMX) were recorded intracellularly in an in vitro slice preparation of the guinea pig brainstem. Bath-applied thyrotropin releasing hormone (TRH) (1-10 microM) induced a reversible depolarization of neurons that was typically accompanied by an increase in the spontaneous firing of the cells. In some cells, TRH induced rhythmic bursting activity. The TRH-induced depolarization occurred also in the presence of reduced Ca2+ and TTX. The response was dose-dependent over TRH concentrations of 0.1-10 microM. The TRH-induced depolarization was accompanied by an increase in input resistance. The reversal potential of this effect corresponded to that of K+. Our results indicate that TRH increases the excitability of DMX neurons by reducing a resting K+ conductance. PMID- 8221079 TI - Spontaneous postischemic hyperthermia is not required for severe CA1 ischemic damage in gerbils. AB - We have recently shown that brain temperature can drop even though rectal and skull readings are maintained near 37 degrees C during global forebrain ischemia in the gerbil. In this study gerbils were subjected to 5 min of ischemia followed by 85 min of extended halothane anesthesia, while rectal and skull temperatures were kept at normal values. This extended anesthesia procedure prevented the development of spontaneous postischemic hyperthermia. However, it occasionally produced mild brain hypothermia both during ischemia and throughout anesthesia. In addition, the degree of brain hypothermia positively correlated with CA1 preservation; with some gerbils showing complete protection. In contrast, animals with normal brain temperature displayed extensive CA1 cell loss. These data suggest that postischemic hyperthermia is not a prerequisite for extensive CA1 loss in gerbils exposed to 5 min of ischemia. Second, rectal and skull recordings are not always reliable indicators of brain temperature, especially during anesthesia. PMID- 8221080 TI - The vascular response to nerve crush: relationship to Wallerian degeneration and regeneration. AB - The response of the endoneurial vasculature in rat sciatic nerve following crush injury was investigated by morphometric analysis of serial nerve transverse sections at the site of injury and in distal segments at 1, 2, 3, 6, and 9 weeks after injury. Quantitative analysis included determination of the number of vessels, vessel radius vessel perimeter, and transfascicular area. The vascular response to crush injury consisted of two phases: an early phase, which peaked at 1 week after crush, consisted of an increase in vessel size but not vessel number. The second phase, which peaked at 6 weeks after crush, consisted of an increase in the number of vessels and in their density. This two-phase response was also evident as a dual peak in the total endoneurial vessel perimeter, a measure of vascular surface area, when this variable was plotted against time. The first phase of the vascular response was temporally related to the recruitment of macrophages and the clearance of degenerating axonal and myelin tissue during the early phase of Wallerian degeneration. The second phase involved an increase in the number of blood vessels and was associated with cellular proliferation, neurite elongation, and myelination during the subsequent period of nerve regeneration. PMID- 8221081 TI - Evidence for the involvement of excitatory amino acid pathways in the development of precipitated withdrawal from acute and chronic morphine: an in vivo voltammetric study in the rat locus coeruleus. AB - Previous studies have demonstrated that activation of excitatory amino acid (EAA) pathways projecting to the locus coeruleus may be involved in the increased firing of locus coeruleus (LC) neurons during opioid withdrawal. Using differential normal pulse voltammetry to monitor catechol oxidation current (CA.OC), an index of neuronal activity in the LC, the role of EAA pathways in naloxone precipitated withdrawal after acute and chronic morphine treatment was examined. Acute morphine treatment (10 micrograms i.c.v.) significantly reduced the CA.OC signal in the LC to 54.3 +/- 3.1% of baseline. Naloxone challenge (1 mg/kg i.v.) completely reversed the morphine effect and produced a significant increase in the CA.OC signal above baseline, peak 145.4 +/- 10.1% of baseline. This naloxone-induced rebound response was attenuated by pretreatment with the EAA receptor antagonists gamma-D-glutamylglycine (DGG) (2, 20, 200 micrograms i.c.v.) and (-)-2-amino-7-phosphonoheptanoic acid (D-APH), but not L-APH (25 micrograms i.c.v.). In chronically morphine-treated rats (25 micrograms/h i.c.v., 5 days), naloxone challenge (1 mg/kg i.v.) produced a significant increase in CA.OC signal, peak 466.5 +/- 112.7% of baseline. This naloxone-induced response was attenuated by pretreatment with DGG (200 micrograms i.c.v.) or D-APH (25 micrograms i.c.v.). To the extent that CA.OC reflects locus coeruleus neuronal activity, the present findings further suggest that increases in locus coeruleus activity during naloxone precipitated withdrawal after both acute and chronic morphine treatment are mediated at least in part by activation of EAA pathways. PMID- 8221082 TI - Degeneration of vascular muscle cells in cerebral amyloid angiopathy of Alzheimer disease. AB - In cerebral amyloid angiopathy, the amyloid-beta (A beta) deposits lie primarily in the tunica media suggesting that smooth muscle cells play an important role in A beta deposition. To define this role, we conducted an immunocytochemical study of brain tissue from cases of Alzheimer disease with extensive cerebral amyloid angiopathy and cerebral hemorrhage. Antibodies specific to recombinant beta protein precursor (beta PP) and synthetic peptides homologous to various beta PP sequences from residue 18 to 689 of beta PP695 were used. Antibodies to actin, tropomyosin, alpha-actinin or desmin were used to label muscle cells. Antibodies to A beta sequences intensely recognized the extracellular amyloid deposit. Antibodies raised against beta PP sequences other than the A beta domain recognized smooth muscle cells. beta PP-immunoreactivity was reduced in regions of A beta deposits, since no muscle cells were recognized by cytoskeletal markers or observed ultrastructurally. In order to assess why A beta is deposited in the tunica media, we used biotin-labelled beta PP to determine if beta PP can be locally retained. We found beta PP bound to the tunica media of vessels but not other brain elements. These findings suggest A beta in blood vessels derives from degenerating beta PP-containing smooth muscle cells. PMID- 8221083 TI - Distribution of phosphatase inhibitor-1-immunoreactive neurons in the suprachiasmatic nucleus of the Syrian hamster. AB - The protein phosphatase inhibitor-1 (I-1) is phosphorylated by a cyclic AMP dependent protein kinase, and is itself involved in the regulation of phosphorylation of other proteins. The enzyme has been shown to be present in skeletal muscles and in distinct neuronal systems of the brain. The suprachiasmatic nucleus is essential in generation of circadian rhythms, but the cellular mechanisms by which the oscillator is entrained are not understood. Since cyclic AMP is known to phase shift the rhythm of electrical activity in SCN neurons in vitro, we aimed by an avidin-biotin immunohistochemical technique to localize I-1-containing neurons in the hamster suprachiasmatic nucleus and thereby identify potential target neurons for cyclic AMP effects. Numerous densely stained neurons were observed in the hamster SCN. The I-1-immunoreactive cell bodies were intermingled with non-immunoreactive neurons and occupied mostly the ventral half of the nucleus, but cell bodies were found in all compartments of the nucleus. The I-1-immunoreactive neurons located in the ventral SCN sent dendrite-like processes into the underlying optic chiasm, indicating that they are directly innervated from the retina, the intergeniculate leaflet of the thalamus, and/or the dorsal raphe. A few I-1-immunoreactive neurons were observed immediately outside the borders of the SCN, but their pronounced staining intensity and their similar morphology to those found inside the SCN indicate that they belong to the same type of neurons as found in the SCN. Delicate I-1 immunoreactive nerve fibers possessing boutons were found throughout the SCN. Furthermore, axonal fibers were followed dorsally into the subparaventricular area.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221084 TI - Increase in inositol tris-, pentakis- and hexakisphosphates by high K+ stimulation in cultured rat cerebellar granule cells. AB - Effects of high K+ stimulation on inositol polyphosphate accumulations and intracellular free calcium concentration ([Ca2+]i) were investigated in cultured rat cerebellar granule cells. When the [3H]inositol-labelled cells were stimulated with KCl, concentration-dependent accumulations of [3H]Ins(1,4,5)P3, [3H]InsP5 and [3H]InsP6 were observed. Nifedipine (3 microM), a calcium channel antagonist, inhibited the high (KCl, 90 mM) K(+)-induced accumulations of these inositol polyphosphates. In Ca(2+)-depleted and EGTA-containing (0.1 mM) medium, the high K(+)-induced inositol polyphosphate accumulation were completely inhibited. Similar results were also observed in the case of [Ca2+]i. These results suggest that the rise in [Ca2+]i caused by activation of voltage dependent calcium channels plays an important roles in the high K(+)-induced accumulation of [3H]Ins(1,4,5)P3, [3H]InsP5 and [3H]InsP6 in cultured rat cerebellar granule cells. PMID- 8221085 TI - The effects of 6-hydroxydopamine lesions of the nucleus accumbens and the mesolimbic dopamine system on oral self-administration of ethanol in the rat. AB - Rats readily learn to self-administer ethanol using a procedure where ethanol is introduced in the presence of a sweetener. After gradual removal of the sweetener, sufficient quantities of ethanol are self-administered in non fluid-, non food-deprived rats to produce reliable blood ethanol concentrations. Previous studies using this self-administration model have shown that dopamine receptor antagonists injected systemically or directly into the terminal regions of the mesolimbic dopamine system decrease lever pressing for ethanol, suggesting an important role for dopamine in ethanol reinforcement. The purpose of the present study was to test the hypothesis that the mesolimbic dopamine system is a critical substrate for ethanol reinforcement. Results of this study show that 6 hydroxydopamine (6-OHDA)-induced lesions of the mesolimbic dopamine system, sufficient to produce a 93% depletion of dopamine in the nucleus accumbens, an 85% depletion in the olfactory tubercle, an 82% depletion in the frontal cortex and a 78% depletion in the amygdala, failed to alter ethanol self-administration as measured by the total lever presses. However, the 6-OHDA lesion rats showed an altered pattern of responding for ethanol: an increase in the slope of the regression line of cumulative responses vs. time and an increase in the frequency of responding at inter-response intervals of 4-6 and 6-8 s post 6-OHDA lesion; suggesting that this lesion produced a subtle change in motor or attentional function. The results of this study indicate that while the mesolimbic dopamine system may contribute to the reinforcing actions of ethanol, it is not critical for maintaining ethanol reinforcement. PMID- 8221086 TI - Characterization of type A and type B CCK receptor binding sites in rat vagus nerve. AB - We employed quantitative receptor autoradiography to analyze pharmacological properties of 125I-Bolton Hunter cholecystokinin (CCK-8)-labeled binding sites in sections of rat cervical vagus nerve that had been ligated 24 h prior to extraction. Binding densities were detected in segments of nerve proximal and distal to the ligature. Analysis was confined to proximal segments. Saturation and competitive binding studies were carried out using sulphated CCK-8 and two selective CCK receptor antagonists: MK-329, to define type-A (CCKA) binding sites; and, L-365,260, to define type-B (CCKB) binding sites. Sulphated CCK-8 was the most potent inhibitor of vagal 125I-CCK binding (IC50 = 2 nM). Nonlinear curve fitting analysis of the CCK binding data favored the presence of a single class of vagal CCK receptors (KDi = 1 nM). However, both MK-329 (IC50 = 18 nM) and L-365,260 (IC50 = 45 nM) competed for vagal 125I-CCK binding indicating the presence of CCKA and CCKB binding sites. Co-analysis of the antagonist binding data suggested that CCKA and CCKB receptors were transported in equal concentrations within the vagus. MK-329 bound with high affinity to CCKA sites (Ki = 3 nM) and low-affinity to CCKB sites (Ki = 462 nM) while L-365,260 bound with high affinity to CCKB sites (Ki = 10 nM) and low-affinity to CCKA sites (Ki = 775 nM). These same ligands were used to characterize the specificity of 125I CCK binding in the medial and lateral divisions of the nucleus of the solitary tract (NTS), two regions innervated by primary vagal afferents carrying CCK receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221087 TI - Ephedrine: effects on neuromuscular transmission. AB - (-)-Ephedrine has been used in the treatment of patients with myasthenia gravis. To investigate the possible effects of ephedrine on neuromuscular transmission, canine intercostal muscle endplates were studied by microelectrode techniques. At concentrations less than 10(-4) M, ephedrine had no effect on neuromuscular transmission. At a concentration of 10(-4) M, ephedrine increased the quantal content of the endplate potential by 21%. The presynaptic store of acetylcholine quanta available for immediate release was unchanged, but the probability of quantal release was increased by 16%. At this concentration, ephedrine decreased the amplitude of the miniature endplate potential by 38%. In the presence of 10( 3) M ephedrine, the miniature endplate potentials and currents became undetectable. The kinetic properties of the acetylcholine receptor channel were studied by analysis of acetylcholine-induced endplate current noise. At 10(-4) M, ephedrine reduced the channel conductance by 43% but had no effect on the open time. At 5 x 10(-4) M, ephedrine reduced the channel conductance by 84% and increased the open time by 23 percent. PMID- 8221088 TI - Species differences in [125I]interleukin-1 binding in brain, endocrine and immune tissues. AB - There were dramatic species differences in the level of [125I]recombinant human interleukin-1 alpha ([125I]hIL-1 alpha) binding with high levels of binding present in mouse and rabbit tissues, while no specific binding was present in rat and guinea pig tissues. Utilizing [125I]hIL-1 alpha, moderate to high levels of specific binding were observed in EL-4 6.1 cells (representative of Type I IL-1 receptors) and in mouse hippocampus, spleen and testis; however, no specific [125I]hIL-1 alpha binding was present in Raji cells (representative of Type II IL 1 receptors) and in rat tissues. On the other hand, utilizing [125I]hIL-1 beta, high specific IL-1 binding was present in EL-4 6.1 and Raji cells and moderate binding was evident in mouse tissues, whereas specific [125I]hIL-1 beta binding to rat tissues was not detectable. Moreover, no IL-1 binding in rat tissues was observed using [125I]hIL-1 receptor antagonist, [125I]mouse IL-1 beta or the homologous radioligands [125I]rat IL-1 beta or [125I]rat IL-1 receptor antagonist. These data demonstrate that under optimal conditions for labeling Type I or Type II IL-1 receptors, no specific binding is observed in rat tissues suggesting the presence of novel IL-1 receptor(s) in rat tissues. PMID- 8221089 TI - Effects of noise stimulation on cochlear dopamine metabolism. AB - Dopamine (DA) appears to be one of the putative neurotransmitters of the lateral efferent olivocochlear fibers. However, its role in the cochlear physiology remains unknown. In this study, animals were exposed for 1 h to white noise at 70, 90 or 110 dB SPL or were kept in silence conditions. Afterwards, the cochlear content of DA and its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were analyzed using HPLC coupled to electrochemical detection. Cochlear DA concentration decreased with the noise intensity, while cochlear DOPAC and HVA concentrations increased. Males presented higher cochlear DOPAC contents and lower HVA contents than females. This sexual dimorphism could be related to the link between DA and gonadal steroids. Present results show that DA, as other lateral efferent neurotransmitters, is released and metabolized in relationship with the noise stimulation, and suggest that DA could be involved in the modulation of the type I afferent fiber activity. PMID- 8221090 TI - Alpha 1-adrenoceptors in the adult rat barrel field: effects of deafferentation and norepinephrine removal. AB - Norepinephrine (NE), acting on brain adrenoceptors, plays an important role in barrel field neuronal activity and plasticity. For this reason, the distribution of alpha 1- and alpha 2-adrenoceptors in the somatosensory cortex barrel field was studied by autoradiographic techniques in rats undergoing plastic change or NE depletion. In layers IV and V of the cortex, the pattern of alpha 1 adrenoceptors (assessed by [3H]prazosin binding) varied across the barrel field. There was relatively low binding within the barrels themselves, with 21% higher binding in the surrounding septa. alpha 2-Adrenoceptor binding (assessed with [3H]paraminoclonidine) was almost homogeneous across the entire barrel field. Two weeks after noradrenergic deafferentation by unilateral lesioning of the locus coeruleus, there was a 16% upregulation of [3H]prazosin binding. This then returned to control levels of by 8 weeks. Peripheral deafferentation of sensory input to the barrel field produced the opposite effect on alpha 1-adrenoceptors. Unilateral removal of all but the central (C3) vibrissa (which induces plastic changes in the cortical representation of the spared virbrissa) caused a 12% decrease in [3H]prazosin binding in the whole barrel field at 2 weeks after surgery which returned to normal by 8 weeks. Therefore, alpha 1-adrenoceptors in the barrel field of the rat are affected in opposite ways by changes in NE content and afferent sensory input. We hypothesize that alpha 1-adrenoceptor levels are modulated after vibrissectomy through either an indirect reaction to reduced cortical gamma-aminobutyric acid levels, or by a reordering of metabolic priorities during plastic change of the cortical neuronal network. PMID- 8221091 TI - Regional cerebral blood flow changes of cortical motor areas and prefrontal areas in humans related to ipsilateral and contralateral hand movement. AB - The regional cerebral blood flow (rCBF) was measured with positron emission tomography (PET) in ten normal right-handed volunteers with the purpose of comparing rCBF changes related to movements of the dominant (right) and non dominant (left) hand. The hand movement task consisted of sequential opposition of the thumb to each finger. The rCBF measured was compared with a rest state. Movements of the dominant hand and the non-dominant hand, increased CBF significantly in the contralateral motor area (MA) and the premotor area (PMA) with small increases in rCBF in the supplementary motor area (SMA). However, movements of the non-dominant hand also elicited significant ipsilateral increases in rCBF in the MA and PMA (6.3% and 5.0%, respectively). Superior part of the prefrontal area (PFA) of the left hemisphere showed significant CBF increases to both left and right hand movement. Our findings indicate that rCBF changes in the motor areas and the PFA of one hemisphere are not related simply to movement of the contralateral hand. Non-dominant hand movement may in addition require activation of ipsilateral motor areas. That is, there appears to be functional asymmetry in the MA and PFA in humans even in this relatively simple and symmetric motor task. PMID- 8221092 TI - Systemic or intracerebroventricular injection of NMDA receptor antagonists attenuates the antinociceptive activity of intrathecally administered NMDA receptor antagonists. AB - We have previously reported that the response latency in the mouse hot-plate test is affected differently by spinal intrathecal (i.t.) injection of competitive and non-competitive N-methyl-D-aspartate (NMDA) receptor antagonists, in that only the former produces an antinociceptive effect. Since the lipophilic non competitive antagonists will redistribute rapidly from the spinal injection site, it is conceivable that they reach sites where they counteract the spinal antinociceptive effect. In the present study, we have tested this hypothesis by comparing the antinociceptive effect of the competitive NMDA receptor antagonist CGS 19755 and the non-competitive NMDA receptor antagonist MK-801 after i.t., intraperitoneal (i.p.) and intracerebroventricular (i.c.v.) administration as well as after combinations thereof. CGS 19755 injected i.p. or i.c.v. and MK-801 injected i.p. or i.t. attenuated the antinociceptive effect of i.t. injected CGS 19755. Both i.p. and i.c.v. administration of either CGS 19755 or MK-801 dose dependently impaired motor function without producing antinociceptive effects. Thus, the effect of CGS 19755 and MK-801 on the motor system was found to be separate from their antinociceptive effect. In a separate experiment, changes in hind-paw skin temperature were excluded as a possible confounding factor. These findings demonstrate that supraspinal systems can limit the spinal antinociceptive effect of NMDA receptor antagonists. PMID- 8221093 TI - Intranigral injections of SCH 23390 inhibit amphetamine-induced rotational behavior. AB - Rats were given unilateral 6-hydroxydopamine lesions of the nigrostriatal pathway and permanent indwelling cannula were surgically implanted into the non-lesioned side of the brain; cannula were used for direct injections of dopamine antagonists into the pars reticulata region of the non-lesioned substantia nigra. The selective D1 receptor antagonist, SCH 23390, was injected intranigrally at various concentrations (3.0, 1.5, 1.0, 0.6, or 0.3 mM) just prior to an intraperitoneal injection of amphetamine. SCH 23390 dose-dependently inhibited amphetamine-induced rotational behavior with the highest doses completely blocking rotational behavior in some animals. An intranigral injection of the selective D2 receptor antagonist, (-)-sulpiride (1.0 mM), did not produce a significant reduction in amphetamine-induced rotational behavior whereas an equivalent molar concentration of SCH 23390 (1.0 mM) produced a significant 62% reduction in amphetamine-induced rotational behavior. A concentration of SCH 23390 that produced a 50% reduction in rotational behavior when injected directly into the substantia nigra was unable to produce a significant reduction in rotational behavior when injected directly into the striatum. The effects of intranigral injections of SCH 23390 on apomorphine-induced rotational behavior were directly opposite to that observed for amphetamine-induced rotational behavior; contralateral rotational behavior increased relative to baseline measures. These data support the hypothesis that dopamine release in the midbrain may act as a neuromodulator of motor behavior, and that D1 receptors play a functional role in this process. PMID- 8221094 TI - Brain blood flow restoration 'rescues' chronically damaged rat CA1 neurons. AB - Middle aged rats (13 months) were subjected to chronic cerebrovascular insufficiency (CVI) for 9 weeks using a 3-vessel occlusion technique. This CVI injury targets CA1 neuron damage selectively. Three groups of rats had their cerebral blood flow restored after 1, 2 or 3 weeks following CVI by removal of their carotid artery occluders. Another rat group did not undergo deocclusion for the 9 week observation period. Rats were tested for memory acquisition and retention 6 and 9 weeks after CVI using a modified water maze test. At the end of the 9 weeks, cerebral blood flow was measured in the fronto-parietal cortex and rats were killed by fixation-perfusion. Hippocampal morphometry was done to assess the % of damaged CA1 neurons and the density of GFAP-positive hyperplasia and hypertrophy. Results show that restoration of cerebral blood flow 1 and 2 weeks after CVI but not after 3 weeks of CVI, reversed a significant increase in reactive astrocytosis and prevented memory impairment in these deoccluded rats when compared to the non-deoccluded group. It appears from these results that 'neuronal rescue' of CA1 neurons is possible when cerebral blood flow is restored in rats subjected to chronic CVI during a 2 week (but not 3 week) 'window of opportunity'. This chronic brain ischemia model may be useful in screening potential therapy in patients with dementia where spatial memory impairment and hippocampal damage may be manifested. PMID- 8221095 TI - Intravenous administration of inorganic selenium compounds, inhibitors of prostaglandin D synthase, inhibits sleep in freely moving rats. AB - Prostaglandin (PG) D2 has been postulated to be an endogenous sleep-promoting factor. Biosynthesis of PGD2 is catalyzed by PGD synthase (prostaglandin-H2 D isomerase, EC 5.3.99.2), the activity of which is inhibited by inorganic selenium compounds such as SeCl4 and Na2SeO3. We recently examined the effect of intracerebroventricular administration of these selenium compounds on sleep in rats, and demonstrated time- and dose-dependent sleep inhibition. To establish whether this effect of selenium is also produced when the compound is administered systemically, we devised a procedure for intravenous catheterization and examined the effect of these selenocompounds on sleep-wake activity in freely moving rats (n = 35). Each test compound was administered into the inferior vena cava continuously between 11.00 and 17.00 h on the experimental day. SeCl4 time- and dose-dependently inhibited sleep at infusion rates of 5, 7.5, 10 and 20 nmol/microliters per min. During the SeCl4 infusion at 20 nmol/microliters per min, slow-wave sleep and paradoxical sleep were reduced to 63% and 50% of their respective baseline values. Na2SeO3 exhibited a similar sleep inhibition, though Na2SO3 was ineffective. Infusion of SeCl4 at 10 nmol/microliters per min or below produced no consistent changes in the mean brain temperature, or food and water intake during the infusion period. During the nocturnal period subsequent to SeCl4 infusion, sleep was increased by a rebound phenomenon, while a decrease in brain temperature and inhibition of food and water intake dose-dependently occurred. We conclude that systemic administration of these PGD synthase inhibitors has a sleep-reducing potency. PMID- 8221096 TI - Sensory responsiveness of brain noradrenergic neurons is modulated by endogenous brain serotonin. AB - Previous results have indicated that application of serotonin (5-HT) onto noradrenergic locus coeruleus (LC) neurons selectively attenuates the response of these cells to excitatory amino acids (EAAs). Other studies revealed that certain sensory responses of LC neurons are mediated by EAA inputs. We examined the role of endogenous 5-HT in modulating sensory responses of LC neurons that are EAA mediated. LC neurons recorded in rats pretreated with the serotonin (5-HT) depletor, p-chlorophenylalanine (PCPA), exhibited increased responsiveness to electrical stimulation of a rear footpad. Conversely, injection of the 5-HT precursor, 5-hydroxytryptophan (5-HTP), reversed this effect of PCPA and attenuated this sensory response of LC neurons in drug-naive animals. Neither treatment altered the spontaneous discharge rate of LC neurons. These results are consistent with previous findings indicating that 5-HT has potent but selective effects on EAA-mediated responses of LC neurons, and in addition point to a possible functional role for endogenous 5-HT in controlling sensory-evoked LC activity. PMID- 8221097 TI - Neuron-specific enolase increases in cerebral and systemic circulation following focal ischemia. AB - Neuron-specific enolase (NSE) is an isoform of the glycolytic enzyme, enolase, and is found in neurons and neuroendocrine cells. We evaluated cerebral immunohistologic and plasma changes in NSE in rats from 2 h to 15 days following permanent or transient middle cerebral artery occlusion (MCAO). At 1-2 days post MCAO, loss of NSE immunofluorescence from within neurons to the extracellular space was observed in the infarcted areas of all MCAO animals. NSE also was identified intravascularly throughout the brain following MCAO. NSE in plasma was determined by a specific radioimmunoassay. Plasma NSE following permanent or transient MCAO was increased significantly from that observed in controls (2.8 +/ 0.3 ng/ml) beginning at 2 h and persisting for 2.5 days post-MCAO (maximum levels of 8.8 +/- 0.9 to 9.6 +/- 0.5 ng/ml after 6-12 h; P < 0.05, n = 4-9). Quantified contralateral forelimb and hindlimb neurological deficits in these animals were significant and persisted for at least 15 days following MCAO but were not observed following sham surgery. These data suggest that MCAO-induced cortical infarction and neurological dysfunction is associated with neuronal depletion and vascular redistribution of brain NSE resulting in a measurable increase in plasma NSE. Such diffusion of NSE into the cerebral vasculature and systemic circulation from ischemic tissue can be expected to serve as a marker for the incidence of cerebral damage in acute and chronic ischemic brain infarcts. PMID- 8221098 TI - Generation of a monoclonal antibody specific for a new class of minor ganglioside antigens, GQ1b alpha and GT1a alpha: its binding to dorsal and lateral horn of human thoracic cord. AB - We have established a monoclonal antibody, GGR41, specific for a new class of minor gangliosides, such as GQ1b alpha and GT1a alpha, by immunizing mice with a GQ1b-rich ganglioside fraction extracted from bovine brain. Each of those minor gangliosides has been reported to be one of the cholinergic-specific gangliosides (Chol-1). Careful examination of binding specificity of the antibody by both an enzyme-linked immunosorbent assay and immunostaining on thin-layer chromatograms showed that the antibody recognizes three sialyl residues separately attaching to the gangliotetraosyl backbone structure. Immunohistochemical analysis revealed that GGR41 immunostained lamina I and III of dorsal horn and lateral horn of human thoracic cord but motor neurons were not immunostained. Except for negative staining of motor neurons, this distribution is similar to the distribution pattern of staining as reported in rats and humans using a polyclonal antibody against Chol-1. Thus, the antibody obtained in this study should be a useful reagent to study the function of a unique new class of the minor gangliosides. PMID- 8221099 TI - Growth hormone release evoked by electrical stimulation of the arcuate nucleus in anesthetized male rats. AB - Plasma growth hormone (GH) concentrations were measured following electrical stimulation of either the arcuate nucleus or the median eminence in urethane anesthetized male rats. While electrical stimulation of the arcuate nucleus elicited a large pulse of GH secretion, stimulation of the median eminence was relatively ineffective. For stimulation of the arcuate nucleus, the frequency dependence of stimulus-secretion coupling for GH release was investigated by delivering 3 differing patterns of electrical stimulation, each of 2 min duration and containing 1,200 stimulus pulses: 10 Hz continuous; 20 Hz (10 s on/10 s off); and 50 Hz (2 s on/8 s off). To examine the effect of increasing the duration of the 50 Hz stimulus train on evoked GH release, a further three stimulation protocols were also tested: 50 Hz (2 s on/8 s off); 50 Hz (3 s on/7 s off) and 50 Hz (4 s on/6 s off). While evoked GH release (per stimulus pulse) was not significantly different for various frequencies of stimulation, it was greatly potentiated by increasing the duration of 50 Hz stimulus trains. These findings suggest that GH release is not linearly related to the activation of GRF neurons but is strongly facilitated with increases in burst duration. PMID- 8221100 TI - Transitional properties of the mechanically evoked perioral reflex from infancy through adulthood. AB - The organization of motor responses in the orbicularis oris muscle following the delivery of punctate mechanical inputs to vermilion skin of the lips was studied in a group of young infants, school-age children, and adults during periods of voluntary lip muscle activation. A specially designed multi-point array skin contactor, coupled to a position-servo controlled linear motor, was highly effective in driving the early component of the perioral reflex (R1). Overall, the evoked R1 response obtained from the infant was of variable amplitude relative to the children and adults, lacked response specificity, and occurred at a longer latency. This brainstem mediated sensorimotor action appears to take on several characteristics of the adult form by the age of 12. The emergence and maturation of mechanically evoked perioral reflexes is discussed in relation to the acquisition of motor skills, including speech and smiling. PMID- 8221101 TI - Differential effects of specific delta and kappa opioid receptor antagonists on the bidirectional dose-dependent effect of systemic naloxone in arthritic rats, an experimental model of persistent pain. AB - In an attempt to determine the opioid receptor class(es) which underly the two opposing effects of naloxone in models of persistent pain, we tested the action of the selective delta antagonist naltrindole, and that of the kappa antagonist MR-2266 on the bidirectional effect of systemic naloxone in arthritic rats. As a nociceptive test, we used the measure of the vocalization thresholds to paw pressure. The antagonists were administered at a dose (1 mg/kg i.v. naltrindole, 0.2 mg/kg i.v. MR-2266), without action per se but which prevents the analgesic effect of the delta agonist DTLET (3 mg/kg, i.v.) or the kappa agonist U-69,593 (1.5 mg/kg, i.v.) respectively, and does not influence the effect of morphine (1 mg/kg i.v.) or the mu agonist DAMGO (2 mg/kg, i.v.) in these animals. In arthritic rats injected with the delta antagonist, the paradoxical antinociceptive effect produced by 3 micrograms/kg i.v. naloxone was not significantly modified (maximal vocalization thresholds (% of control) were 146 +/- 9% versus 161 +/- 7% in the control group). By contrast, the hyperalgesic effect produced by 1 mg/kg i.v. naloxone was significantly reduced (maximal vocalization thresholds were 87 +/- 4% versus 69 +/- 5% in the control group). In rats injected with the kappa antagonist, the antinociceptive effect of the low dose of naloxone was almost abolished (mean vocalization thresholds were 115 +/- 3% versus 169 +/- 7%) whereas the hyperalgesic effect of naloxone 1 mg/kg i.v. was not significantly modified (mean vocalization thresholds = 70 +/- 3% and 65 +/- 3%, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221102 TI - Glycogen in astrocytes: possible function as lactate supply for neighboring cells. AB - In order to contribute to the elucidation of the function of astrocyte glycogen in brain, studies on the fate of the glucosyl residues of glycogen were carried out on astroglia-rich primary cultures derived from the brains of newborn rats. On glucose deprivation astroglial cells rapidly deplete their glycogen. In contrast to the situation with hepatocytes, only lactate, but not glucose, is detectable in the medium surrounding the astroglial cells. Besides glucose, astroglial cultures can also use mannose as a substrate for the synthesis of glycogen and the generation of lactate. Although mannose-fed astroglial cells contain glucose-6-phosphate, they do not release a measurable amount of glucose into the culture medium. Instead of glucose the astroglial cells release high amounts of lactate into the culture medium. Gluconolactone or 2-deoxyglucose which prevent glycogen breakdown in astroglial cells after glucose deprivation, allow to discriminate between lactate generated from glycogen and lactate from other sources. The amount of lactate found in the medium in the absence of gluconolactone (or 2-deoxyglucose) exceeds the amount found in the presence of either compound by the lactate equivalents calculated to be contained in the cellular glycogen. In conclusion, glycogen in astrocytes can be considered as a store for lactate rather than for glucose. PMID- 8221103 TI - Chlordiazepoxide attenuates stress-induced accumulation of corticotropin releasing factor mRNA in the paraventricular nucleus. AB - Corticotropin-releasing factor (CRF) plays a role in coordinating endocrine, autonomic and behavioral responses to stressful stimuli. Benzodiazepines exert many effects which are antithetical to those of CRF, including anxiolysis and suppression of the pituitary-adrenal axis. Although there is evidence that benzodiazepines can modulate several electrophysiological and behavioral responses to exogenous CRF, we questioned whether this class of drug might also affect CRF biosynthesis as well. We have shown previously that footshock stress increases CRF mRNA levels as monitored by in situ hybridization histochemical techniques in the paraventricular nucleus (PVN) and Barrington's nucleus (the pontine micturition center). We report here the effects of the potent benzodiazepine, chlordiazepoxide (CDP), on stress-induced CRF mRNA accumulation in these two regions. Male albino rats were exposed to electrical footshock (1.5 mA, 1-s duration, 60 times/30 min) twice daily for 4 days and sacrificed 24 h after the last shock session. Either CDP (1, 2.5, 5 or 10 mg/kg) or saline was given i.p. 30 min before each stress. Sections were hybridized with an 35S labeled prepro-CRF cRNA probe. Relative levels of CRF mRNA were quantified by densitometry of the autoradiography with X-ray film. CRF mRNA concentrations were significantly increased in both the PVN and Barrington's nucleus after stress, and CDP attenuated these increases in the PVN. By contrast, CDP did not affect CRF mRNA accumulation in Barrington's nucleus after stress. The results suggest that the benzodiazepine, CDP, suppresses stress-induced pituitary adrenal activation at least in part through inhibition of CRF production in the PVN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221104 TI - Attenuation of stress-induced behavior by antagonism of corticotropin-releasing factor receptors in the central amygdala in the rat. AB - Research suggests that endogenous corticotropin-releasing factor (CRF) in the amygdala plays a role in the expression of stress-induced behavior. This study examined in rats whether antagonism of CRF receptors in the central amygdala (CA) region using alpha-helical CRF9-41, a CRF antagonist, was effective in attenuating the occurrence of stress-induced freezing. Bilateral infusions of 50, 100, or 200 ng of the CRF antagonist were made in the CA region using 33-gauge cannula immediately prior to testing. Freezing was measured in two test conditions. In one condition, the effects of the CRF antagonist on freezing was assessed immediately after exposure to electric foot shock. In the other condition, freezing was examined in shock-experienced rats that were re-exposed to the shock environment. Results suggested that 50 and 100 ng of the CRF antagonist were effective in reducing the duration of freezing in the immediate post-shock period. In addition, the 100 ng dose produced a significant reduction in freezing duration after rats were re-exposed to the shock environment. Collectively, data suggest that antagonizing the action of endogenous CRF in the CA region contributes to a general alleviation of stress-induced freezing. PMID- 8221105 TI - Neurons in the area of human thalamic nucleus ventralis caudalis respond to painful heat stimuli. AB - A population of neurons in the area of human thalamic nucleus ventralis caudalis (Vc) respond to noxious heat stimuli. In the cutaneous core of Vc 6% (6/108) of recorded neurons had a significantly greater response to noxious heat stimuli than to innocuous control stimuli. Half of these neurons (n = 3) also responded to innocuous cold stimuli. Within the region posterior and inferior to the cutaneous core of Vc 5% (4/77) of neurons responded exclusively to noxious heat stimuli. Cells responding to noxious heat were recorded at a greater proportion (66%) of sites where painful sensations were evoked by microstimulation than at sites where nonpainful sensations were evoked (1.5%). The results suggest that neurons in the region of human Vc mediate the sensory aspect of pain. PMID- 8221106 TI - Spontaneous activity in the thalamic reticular nucleus during the sleep/wake cycle of the freely-moving rat. AB - Neurons of the somatosensory thalamic reticular nucleus (TRN) were studied by extracellular recordings through the sleep/wake cycle in the unanesthetized, freely-moving rat. All electrophysiologically-identified TRN neurons expressed rhythmic patterns of discharge that altered with shifts in sleep/wake state. During slow wave (SW) sleep, neurons displayed spike-burst discharges in long trains followed by pauses. high-frequency oscillations in auto-correlograms in the spindle-frequency range (approximately 10 Hz) reflected the rhythm of interburst intervals within the trains whereas low-frequency oscillations (0.3 0.2 Hz) displayed the rhythm of intertrain intervals. During rapid eye movement (REM) sleep, a more continuous pattern of spike-burst discharges was prominent, resulting in absence of a detectable, low-frequency rhythm but persistence of spindle-frequency firing. At the transitions between SW and REM sleep, cell discharge was more tonic than during either sleep state and lacked a dominant rhythm. During the wake (AW) state, neurons fired in a single-spike mode that also lacked rhythmicity. Unlike their pattern of discharge, TRN neurons' mean rate of discharge did not distinguish sleep/wake state. The mean discharge rates were: SW, 18.4 +/- 1.3; REM, 17.4 +/- 1.2; AW, 22.3 +/- 2.1 (Hz +/- S.E.M.). Mean discharge rate during transitions from SW to REM sleep (28.6 +/- 2.1) was significantly higher, however, than during any sleep/wake state. Compelling evidence was lacking for segregation of TRN neurons into discrete populations according to absolute discharge rate. Neurons recorded simultaneously from the same electrode discharged synchronous trains of spike-bursts and pauses during SW sleep. This phenomenon may be related to generation of EEG slow waves.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221107 TI - Functional mapping of neural sites mediating prolactin-induced hyperphagia in doves. AB - Microinjections of prolactin (PRL) into the ventromedial nucleus of the hypothalamus (VMN) or the preoptic area (POA) have been previously shown to increase food intake and body weight in ring doves. In an attempt to corroborate these results and to provide a more complete map of PRL-sensitive brain sites mediating the orexigenic action of PRL, a microinjection procedure was employed in the present study that delivered PRL or saline vehicle in extremely small volumes (10 nl/injection) to a variety of diencephalic sites in dove brain that had been previously demonstrated to contain high concentrations of PRL receptors. Estimates obtained from one female subject given a single 10 nl injection of [125I]ovine PRL into the VMN supported the claim that such injection volumes resulted in limited diffusion, as 80% of the tissue radioactivity was found within a 280 mm area surrounding the injection site at 30 min after injection. Food intake of cannulated male doves in the mapping study was monitored daily during a 6 day baseline period, an initial 4 day treatment period, a 6-12 day post-treatment recovery period, and a second 4 day treatment period. Approximately half of the birds received PRL injections (50 ng/10 nl twice daily) and saine vehicle injections (10 nl twice daily) during the first and second treatment periods, respectively, while remaining birds received these treatments in the reverse order. No significant changes in food intake across baseline, vehicle, post-treatment, or PRL treatment periods were observed in birds with injection sites in the lateral POA, paraventricular nucleus of the hypothalamus (PVN), or the medial-basal hypothalamic region between the tuberal hypothalamus (TU) and VMN. In contrast, injections of PRL into the VMN area, medial POA, or TU resulted in average daily food intake values that significantly exceeded those recorded during other periods. The most robust feeding response was seen in the VMN group, where PRL injections resulted in a 58% increase in food intake over that recorded during injection of vehicle. This increase was significantly greater than that observed following PRL injections into the mPOA (26%) or the TU (32%). These findings suggest that the VMN may be a primary site of PRL action in promoting hyperphagia in this species, although PRL effects at other diencephalic loci, such as the mPOA and TU, may also contribute to the orexigenic action of this hormone. PMID- 8221108 TI - Identification of periaqueductal gray and dorsal raphe nucleus neurons projecting to both the trigeminal sensory complex and forebrain structures: a fluorescent retrograde double-labeling study in the rat. AB - The midbrain periaqueductal gray (PAG) including the dorsal raphe nucleus (DR) has been known to contain serotoninergic neurons projecting to many brain regions. Employing fluorescent retrograde double labeling combined with immunofluorescence histochemistry for serotonin (5-HT), we examined in the rat whether or not single PAG/DR neurons with 5-HT send their axons to both the trigeminal sensory complex and forebrain regions. Stereotaxic injections of Diamidino Yellow (DY) and Fast Blue (FB) were performed unilaterally; DY was injected into the caudal spinal trigeminal nucleus or principal sensory trigeminal nucleus, and FB into the ventrolateral orbital cortex, nucleus accumbens or amygdala. A small percentage of PAG/DR neurons were doubly labeled with DY and FB, and the majority of them showed 5-HT-like immunoreactivity (5-HT LI). Most of these 5-HT-LI PAG/DR neurons that were indicated to send their axons simultaneously to both the trigeminal sensory complex and forebrain regions were distributed in the ventrolateral PAG subdivision and ventral aspects of the medial PAG subdivision at the middle and caudal PAG levels, bilaterally with a predominant distribution on the side ipsilateral to the injections. This indicates a possible role of these PAG/DR neurons in the limbic or affective motivational aspect of the pain-related neural system. PMID- 8221109 TI - Neurons in the superficial dorsal horn of the rat spinal cord projecting to the medullary ventrolateral reticular formation express c-fos after noxious stimulation of the skin. AB - The nociceptive nature of the neurons of the superficial dorsal horn (laminae I III) which project to the medullary ventrolateral reticular formation is studied in the rat. Medullary injections of Fluoro-Gold showed exclusive retrograde labeling of laminae I-III cells when the tracer filled a zone intermediate between the lateral tip of the lateral reticular nucleus and the spinal trigeminal nucleus, pars caudalis. This zone is here called VLMlat. Following noxious mechanical or thermal stimulation of the skin, double-labeled neurons, which stained retrogradely and were Fos-immunoreactive, prevailed in laminae I and IIo. Double-labeled neurons were few in lamina IIi after thermal stimulation and entirely lacking in lamina III after the two kinds of stimulation. Findings in lamina I confirm previous electrophysiological data (see Menetrey et al., J. Neurophysiol., 52 (1984) 595-611) showing that lamina I cells projecting to the ventrolateral reticular medulla convey noxious messages. The occurrence of numerous double-labeled cells in lamina IIo suggests that this lamina is also involved in nociceptive transmission to the VLMlat. PMID- 8221110 TI - High metabolic activity demonstrated by positron emission tomography in human auditory cortex in case of deafness of early onset. AB - Glucose metabolism has been studied in the auditory cortex of human subjects with deafness of early onset, and compared to normal subjects with ears plugged. The metabolism in the auditory cortex and in the association auditory cortex was higher in deaf subjects than in normal subjects. This result is compared to similar observations that we made previously in the visual cortex of human subjects with blindness of early onset. PMID- 8221111 TI - An analysis of GABAergic afferents to basket cell bodies in the cat's cerebellum. AB - An antibody to glutamic acid decarboxylase (GAD) was used to identify GABAergic terminals around the somata of basket cells in the cat's cerebellar cortex. Two sources for GAD immunoreactive terminals were identified based on size and cytological characteristics including the recurrent collaterals derived from Purkinje cells and the axons of stellate cells. The majority of the unlabeled terminals likely arise from parallel fibers. The present analysis indicates that GAD-positive terminals form 13% of the synaptic contacts on basket cells. The remaining 87% are unlabeled. Thus, GAD-positive terminals represent a small proportion of the synaptic input to basket cell bodies as compared to afferent endings that likely mediate excitation. PMID- 8221112 TI - Loss of sigma binding sites in the CA1 area of the anterior hippocampus in Alzheimer's disease correlates with CA1 pyramidal cell loss. AB - The densities of [3H]1,3-di-o-tolylguanidine ([3H]DTG) binding to sigma binding sites in the CA1 stratum pyramidale region in 7 hippocampi affected by Alzheimer's disease, were compared with densities in 7 normal hippocampi. There was an average reduction of 26% in [3H]DTG binding in this area, which was correlated with an average 29% pyramidal cell loss in the same region. These results are consistent with experiments in animals indicating that sigma binding sites are preferentially associated with the somata of large cells. PMID- 8221113 TI - L-dopa pretreatment potentiates striatal dopamine overflow and produces dopamine terminal injury after a single methamphetamine injection. AB - Rats receiving L-dopa/carbidopa (70 mg/kg/17.5 mg/kg, i.p.) 1 h prior to a single methamphetamine (m-AMPH) (4 mg/kg, s.c.) pretreatment showed an extraordinary striatal dopamine (DA) overflow into the extracellular space (30-60 times basal overflow) as compared to the DA overflow elicited by m-AMPH alone (4-5 times basal). Animals treated with L-dopa/carbidopa plus m-AMPH, but not m-AMPH alone, had substantial (60%) decreases in striatal DA content 1 week later. These findings support the conclusion that the magnitude of m-AMPH-induced DA overflow contributes to the degree of nerve terminal damage and highlight the importance of extracellular DA in striatal terminal damage. PMID- 8221114 TI - Attenuation of cholinergic analgesia by nifedipine. AB - Nociception was tested in mice receiving oxotremorine or physostigmine either after the dihydropyridine calcium channel blocker nifedipine or the non-calcium antagonist vasodilator hydralazine. Nifedipine did not change the reaction time to thermal stimulation (tail-flick test), but attenuated the prolonging action on tail-flick latencies exerted by the two cholinomimetic agents. Hydralazine had no effect alone nor modified the action of cholinomimetics. The results suggest that attenuation of cholinergic analgesia by nifedipine might be related to not yet defined neuronal changes produced by calcium channel blockade, but changes in the pharmacokinetics of oxotremorine and physostigmine cannot be ruled out. PMID- 8221115 TI - Multiple opioid system involvement in the mediation of parasitic-infection induced analgesia. AB - Although parasite modification of host behaviour is well established, little is known about the mechanisms underlying such effects. The present study examined the relationships between subclinical infection with the enteric sporozoan parasite, Eimeria vermiformis, nociceptive responses and endogenous opioid systems in male mice. Infected mice displayed significant analgesia which increased through the prepatent period [oocyst formation (pre-infective); days 1 7 post-infection (PI)], reached a maximum with the onset of patency (onset oocyst shedding and infectivity; days 7-8 PI) and declined during patency (oocyst shedding), with response latencies declining to basal levels with the cessation of oocyst production and infectivity (day 15 PI). The increasing nociception during the prepatent period (day 4 PI) was associated with kappa opioid mechanisms, being reduced by the kappa antagonist, nor-binaltorphimine, and insensitive to either the delta antagonist, ICI 174,864, or the general, predominantly mu antagonist, naloxone. Maximum analgesia (day 7 PI) associated with the onset of patency (infectivity) was sensitive to both the kappa and mu antagonists, but insensitive to the delta antagonist, while the declining analgesia during patency (day 10 PI) was reduced by the mu and delta antagonists, but was insensitive to the kappa antagonist. These results indicate that mu, delta and kappa opioid systems are involved in the mediation of subclinical parasitic infection-induced analgesia and likely other associated parasite induced modifications of host behaviour. PMID- 8221116 TI - Interleukin-1 mediates the behavioral hyperalgesia produced by lithium chloride and endotoxin. AB - The sickness-inducing agents lithium chloride (LiCl) and lipopolysaccharide (LPS) produce a long-lasting facilitation of the nociceptive tailflick reflex. Many of the behavioral and physiological changes produced by illness are mediated by interleukin-1 (IL-1) released from monocytes stimulated by the pathogenic substance. Monocytes also produce an IL-1 receptor antagonist (IL-1ra) which has been sequenced and cloned. The present experiments report that IL-1 can itself produce hyperalgesia as assessed by tailflick to radiant heat, and that recombinant IL-1ra blocks the hyperalgesia produced by LiCl and LPS. PMID- 8221117 TI - L-dopa stimulates c-fos expression in dopamine denervated striatum by combined activation of D-1 and D-2 receptors. AB - Administration of L-dopa to unilaterally 6-hydroxydopamine-lesioned rats, activates the early gene c-fos in the lesioned caudate-putamen. D-1 receptor blockade by SCH 23390, prevented L-dopa-induced Fos-like immunoreactivity in the whole caudate-putamen, while D-2 receptor blockade by raclopride reduced Fos-like immunoreactivity only in the dorso-lateral portion. The results suggest that L dopa induces c-fos primarily through an activation of D-1 receptors, while D-2 receptor stimulation plays a facilitatory influence on D-1-mediated c-fos expression. PMID- 8221118 TI - Antibodies to quinolinic acid reveal localization in select immune cells rather than neurons or astroglia. AB - Polyclonal antibodies were produced against quinolinic acid. No immunoreactivity was observed in any cell type in carbodiimide-fixed brain tissue from control rats. When the antibodies were applied to carbodiimide-fixed spleen tissue, strong quinolinic acid immunoreactivity was observed in some cells with the appearance of macrophages and dendritic cells. These findings indicate an immune system origin for quinolinic acid, and implicate immune cells in excitotoxic CNS pathologies. These findings also raise the possibility that quinolinic acid is a unique cytokine in immune system signal transmission. PMID- 8221119 TI - Individual differences in the psychomotor effects of morphine are predicted by reactivity to novelty and influenced by corticosterone secretion. AB - Clinical observations show that individual vulnerability to the reinforcing properties of drugs plays an important part in the subsequent development of addition. In animals, individual vulnerability to psychostimulants has been found to be predicted by their locomotor response to novelty as well as their corticosterone response. Rats with a high locomotor response to novelty (High Responders or HR) relative to Low Responders (LR), show a higher sensitivity to both the psychomotor and reinforcing effects of psychostimulants and a longer lasting corticosterone secretion in response to stress. In this study, we addressed two main questions. First, does the locomotor response to novelty also predict the psychomotor effects of morphine? Second, do differences in corticosterone secretion underlie individual differences in the stimulant effects of morphine? We compared the locomotor response to morphine (2 mg/kg s.c.) in: (i) HR and LR rats with an intact hypothalamo-pituitary-adrenal (HPA) axis; (ii) HR and LR rats in which stress-induced corticosterone secretion was suppressed by adrenalectomy but basal levels of corticosterone were maintained by implantation of subcutaneous corticosterone pellets. In animals with an intact HPA axis, HR rats showed a higher locomotor response than did LRs to morphine. In animals in which corticosterone secretion was suppressed, the enhanced locomotor response of the HRs to morphine fell to that observed in the LRs. In conclusion our data show that, (1) individual reactivity to novelty can predict individual vulnerability to the psychomotor effects of opioids, and (2) stress-induced corticosterone secretion may play a role in determining individual differences in sensitivity to these drugs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221120 TI - Increase of central 5-HT1B binding sites following 5,7-dihydroxytryptamine axotomy in the adult rat. AB - The effects of selective axotomy of serotoninergic neurons produced by an intracerebroventricular injection of 5,7-dihydroxytryptamine (200 micrograms free base) on 5-HT1B binding sites labeled with S-CM-G-[125I]TNH2 were investigated by quantitative autoradiography in the rat brain. Results show, 21 days after surgery, an upregulation of 5-HT1B receptors in the entorhinal cortex and the dorsomedial and suprachiasmatic nuclei of the hypothalamus. The cellular localization of those 5-HT1B receptors exhibiting post-lesion plastic properties is discussed. PMID- 8221121 TI - Hypoxia affects differently the intracellular pH of clustered and isolated glomus cells of the rat carotid body. AB - Clustered and isolated glomus cells, cultured from rat carotid bodies, were exposed to hypoxia (pO2 2-30 torr) induced by applications of sodium-dithionite (Na2S2O4). Hypoxia decreased or increased intracellular pH (pHi) of clustered cells about equally, but lowered it in most isolated cells. The levels of intracellular acidification were similar in both groups whereas alkalinization was more pronounced in the clusters. The H+ equilibrium potential (EH) and its changes during hypoxia (delta EH), were determined almost exclusively by pHi. Seventy-five percent of clustered cells became depolarized whereas 80% of isolated cells underwent hyperpolarization. In both groups, changes in the resting potential (delta EM) were directly and significantly correlated with delta EH, thus delta pHi. These observations support the view that clustered and isolated rat glomus cells behave differently. This difference may occur because of the presence (in the clusters) or absence (in isolated cells) of enveloping sustentacular cell processes. PMID- 8221122 TI - Morphology of CA3 neurons in hippocampal slices with nonepileptic and epileptic activity: a light and electron microscopic study. AB - In guinea pig hippocampal slices, relations between morphology and spontaneous bioelectric activity of neurons were studied in control saline and with exposure to the epileptogenic drug pentylenetetrazole (PTZ) for 2-3 h. Light and electron microscopic structures of the CA3 region were analysed after recording the membrane potential. Neurons in slices kept in control saline exhibited spontaneous aperiodic bioelectric activities partly mixed with rhythmically occurring burst discharges. In slices exposed to PTZ, these periodic burst discharges and/or paroxysmal depolarization shifts (PDS) predominated. Light microscopic comparison focussing on tissue preservation showed no significant differences between control and PTZ-treated slices. Ultrastructural morphology revealed, on the one hand, no differences regarding spine and synaptic densities, and on the other hand, significantly more irregular electron translucent vacuoles within dendrites of PTZ-treated slices being either randomly distributed or clustered. The vacuoles are interpreted as early changes during epileptic activity. PMID- 8221123 TI - Scopolamine injected into the rat amygdala impairs working memory in the double Y maze. AB - Recent neurochemical results suggest the hypothesis that the nucleus basalis magnocellularis (nbm) cholinergic projection to the amygdala may play a role in memory. The present study investigated the effects of intra-amygdaloid injections of the cholinergic antagonist scopolamine on working and reference memory in the double Y-maze. Rats were pretrained until working and reference memory choice accuracy stabilized to a criterion of > or = 86% correct. Bilateral cannulae were then surgically implanted in the basolateral amygdaloid complex. Rats (n = 9) received scopolamine in doses of 8.0, 24.0, and 72.0 micrograms/0.5 microliter and saline (0.5 microliter) in a counterbalanced order with retraining to criterion between injections. Intra-amygdaloid scopolamine produced a dose dependent and differential impairment of working and reference memory. A dose of 24.0 micrograms impaired working memory without significantly affecting reference memory; doses of 8.0 micrograms and 72.0 micrograms affected neither and both types of memory, respectively. Results implicate amygdaloid acetylcholine in memory. PMID- 8221124 TI - Fornix degeneration and memory in traumatic brain injury. AB - Fornix-to-brain ratios (FBR) based on postinjury magnetic resonance (MR) studies were calculated on a group of 27 female traumatic brain injury (TBI) patients and 18 female medical controls by taking the widest aspect of the fornix at the level of the anterior horns and third ventricle and determining a fornix surface area. The FBR was significantly reduced in the TBI group (FBR = 0.1121) as compared to the normal control group (FBR = 0.1766). Despite these significant FBR findings indicating prominent atrophic changes of the fornix in TBI patients, the FBR did not relate systematically to neuropsychological outcome. These findings clearly indicate fornix vulnerability in TBI and that current quantitative MR methods are sensitive enough to detect such changes. However, fornix degeneration constitutes only one of many contributing factors to the anatomic basis of TBI-induced cognitive disturbances, as fornix atrophy did not relate systematically to neuropsychological outcome. PMID- 8221125 TI - Glucocorticoids regulate the development of intracellular signaling: enhanced forebrain adenylate cyclase catalytic subunit activity after fetal dexamethasone exposure. AB - Although glucocorticoids cause growth retardation and interfere with cell development, selective promotion of some aspects of cell function also has been reported. The current study examines whether glucocorticoids enhance intracellular transduction mechanisms mediated by adenylate cyclase in the developing forebrain, a region in which steroids have been shown to interfere with cell replication, maturation, and growth. Pregnant rats were given dexamethasone at doses spanning the threshold for growth impairment (0.05, 0.2, and 0.8 mg/kg) on gestational days 17, 18, and 19, and development of adenylate cyclase was evaluated in membrane preparations, using four different activity measures; basal adenylate cyclase in the absence or presence of GTP, maximal G protein activation by fluoride in the presence of GTP, and stimulation mediated by forskolin-Mn2+, which bypasses the G-proteins. Prenatal exposure to dexamethasone produced a dose-dependent impairment of body growth, with smaller deficits in forebrain weights (brain sparing) indicative of systemic toxicity. Basal adenylate cyclase activity was unaffected by dexamethasone treatment, regardless of whether GTP was present in the assay. Similarly, fluoride stimulation developed normally in all dexamethasone groups. However, forskolin Mn(2+)-stimulated activity was significantly enhanced in a dose-dependent fashion. These results suggest that glucocorticoids serve as positive factors for the development of adenylate cyclase catalytic subunit activity, independently of their adverse effects on general growth and development; thus, these hormones may be a primary regulator of cell signaling during early development. PMID- 8221126 TI - The pathway responsible for EEG synchronization and effect of histamine on this system. AB - Electrical stimulation (3 Hz, 0.5 volts) to the midbrain reticular formation of conscious rats induced significant increase of EEG power densities (synchronization) recorded at the frontal cortex (FCOR), nucleus ventralis thalami (VE), or nucleus medialis centralis thalami (CM). Significant synchronization was also observed in the FCOR when electrical stimulation was applied to the VE and CM. When ipsilateral and bilateral VEs were electrocoagulated, no EEG synchronization was observed in the FCOR and CM. Intracerebroventricular administration of histamine (Hi) caused a marked suppression of FCOR EEG synchronization in both CM-lesioned and normal rats through H1 receptors. EEG synchronization in FCOR was not induced in ipsilateral or bilateral VE-lesioned rats after RF stimulation. When Hi (1 microgram) was injected into the VE of normal rats, EEG synchronization of FCOR was markedly reduced after RF or VE stimulation. No such changes were induced when Hi was injected into the CM. PMID- 8221127 TI - Neurotensin-induced hypothermia prevents hippocampal neuronal damage and increased locomotor activity in ischemic gerbils. AB - Hypothermia induced by surface cooling has shown to protect vulnerable regions of the brain during an ischemic insult. This study evaluated the neuroprotective efficacy of neurotensin, a potent hypothermic agent, using a 5-min carotid occlusion procedure in the gerbil. In Experiment 1, the dose-response and time course of neurotensin-induced hypothermia were evaluated (n = 5/dose). Central infusion of 10, 20, and 30 micrograms neurotensin were found to significantly decrease core body temperature of conscious gerbils within 30 min of administration. In Experiment 2, gerbils pretreated with 30 micrograms neurotensin were permitted to become hypothermic or were maintained at 37 degrees 38 degrees C (rectal) during ischemic insult. Other gerbils were pretreated with peptide vehicle prior to ischemic insult (at 37 degrees -38 degrees C) or underwent a sham procedure (n = 6/condition). At 24 h after surgery, gerbils were tested for increased locomotor activity in an open-field apparatus. Gerbils pretreated with peptide vehicle or neurotensin and maintained at 37 degrees-38 degrees C during ischemia had significantly higher activity levels compared to the other treated groups. In contrast, gerbils made hypothermic with neurotensin exhibited activity levels similar to sham gerbils. Histological assessment revealed that neurotensin-induced hypothermia protected the CA1 region from ischemic damage. PMID- 8221128 TI - Elevation of hippocampal extracellular acetylcholine levels by methoctramine. AB - Previous studies suggest that m2 muscarinic receptors serve as presynaptic autoreceptors. Blocking these receptors by selective antagonists may, therefore, lead to increased acetylcholine (ACh) release. This study assessed changes in extracellular ACh levels, via in vivo microdialysis, following administration of the m2 antagonist methoctramine. Drug or placebo (Ringer's solution) was perfused via a microdialysis probe into the CA1 hippocampal region of unrestrained, awake male Fischer rats. HPLC-EC was used for online analysis of the dialysates. Methoctramine significantly enhanced ACh release in a dose-dependent fashion as compared to placebo for the doses employed (0.25-16 microM). The present in vivo data corroborate studies that show increased ACh levels in vitro following application of m2 antagonists. PMID- 8221129 TI - Brain angiotensin AT-2 receptor antagonism and water intake. AB - Intracerebroventricular administration of the angiotensin AT-2 receptor antagonist, PD 123319, inhibited drinking induced in rats by hypertonic NaCl, carbachol, isoproterenol, hypovolemia, and water deprivation, but had no effect on food intake. In contrast, the AT-1 antagonist, losartan potassium, had no effect on these intakes. A model of thirst is presented that incorporates an AT-2 receptor in a final common pathway for drinking. PMID- 8221130 TI - Streptozotocin-induced diabetes blocks the positive feedback release of luteinizing hormone in the female rat. AB - The effects of streptozotocin-induced (STZ) diabetes on the release of gonadotropins was studied in female rats. In the first experiment, rats were ovariectomized and 2 days later were injected with STZ. Three weeks later rats were treated with estrogen and progesterone and blood samples were taken via intraatrial cannulae for luteinizing hormone (LH) assay. Afternoon surges of LH were seen in 4/5 control but 0/8 STZ rats. Pituitary responses to LH-releasing hormone in vitro did not differ. In the 2nd experiment, ovariectomized estrogen primed rats were killed prior to and during a progesterone-induced LH surge. As in Experiment 1, STZ-treatment inhibited the LH surge but did not effect the afternoon rise in median eminence norepinephrine turnover which has previously been shown to be important in stimulating LH release. Turnover of norepinephrine in the anterior hypothalamus was depressed in the diabetic rats both prior to and during the expected time of the LH surge. Dopamine turnover was depressed in all three brain regions studied. It can be concluded that the positive feedback control of LH release is severely attenuated in diabetic rats but the mechanism explaining the loss is not clear. Diabetes-induced alterations in hypothalamic catecholamine metabolism may be involved but further work is needed to more carefully define these relationships. PMID- 8221131 TI - Angiotensin II binding in area postrema and nucleus tractus solitarius of SHR and WKY rats. AB - The distribution of angiotensin II (AII) binding sites in the area postrema (AP) and adjacent nucleus tractus solitarius (NTS) was compared in spontaneously hypertensive (SHR) rats and normotensive Wistar-Kyoto (WKY) rats. 125I[Saralasin 1-Isoleucine8]-Angiotensin II (125I[SI]-AII) binding density was quantitated from autoradiographic images by computer-assisted image analysis. Seventeen 30 microns serial coronal sections inclusive of the entire AP were analyzed as either individual sections or as groups of sections designated as caudal, middle, or rostral area postrema regions. 125I[SI]-AII binding density was greatest in caudal AP and declined progressively in the rostral direction in both strains; however, binding density in SHR was significantly higher than in WKY rats at each level of the AP analyzed. 125I[SI]-AII binding in the entire area postrema was approximately 46% higher in SHR rats. In the NTS, however, there were no differences in binding density between strains. At the middle level of the AP, 125I[SI]-AII binding was highest in the ventral midline and lowest in the dorsolateral region. In the NTS, the highest 125I[SI]-AII binding density was found in the pars commissuralis and pars medialis. In conclusion a) 125I[SI]-AII binding density was nonhomogeneous in the AP and NTS of both strains, b) qualitatively similar patterns of nonhomogeneity of binding in the AP and NTS were noted in both strains; however, c) the SHR strain consistently had higher density AII binding than WKY in AP, but not in NTS. PMID- 8221132 TI - The effect of insulin on norepinephrine uptake by PC12 cells. AB - We have previously reported that insulin can enhance endogenous noradrenergic activity in vitro in the rat CNS. In the present study, we examined one potential mechanism underlying this effect: the ability of insulin to inhibit norepinephrine (NE) reuptake and secondarily increase its synaptic concentration. Acute (20 min) insulin treatment (0.1-10 nM) significantly inhibited specific 3H norepinephrine uptake by rat hypothalamic slices. To ascertain whether this is a direct effect of insulin on cells that can synthesize and release norepinephrine, we studied NE uptake by the rat pheochromocytoma PC12 cell line. In PC12 cells, insulin (0.5-10 nM) inhibited NE uptake whereas the related peptide, insulin-like growth factor 1 (IGF-1), did not. Insulin did not compete with 3H-mazindol (a ligand for the NE reuptake transporter) binding to PC12 cell membranes. Thus, this effect of insulin is not due to interaction with either IGF-1 receptors or the norepinephrine transporter, but may be due to insulin interaction with its own receptor. Chronic (96-h) insulin treatment of PC12 cells also resulted in an inhibition of 3H-norepinephrine uptake, and membranes prepared from cells chronically treated with insulin bound less 3H-desipramine than control membranes. Thus, chronic insulin treatment may result in a decrease in the numbers of membrane-associated transporters. We conclude that insulin has a direct and physiological role in the modulation of synaptic norepinephrine levels by modulating reuptake by cells that synthesize and release norepinephrine. PMID- 8221133 TI - Melatonin reduces dopamine content in the neurointermediate lobe of male Syrian hamsters. AB - The effect of daily late afternoon administration of melatonin on the in situ activity of tyrosine hydroxylase (TH) was studied in the posterior pituitary (neurointermediate lobe) of the male Syrian hamster. After 3 weeks of melatonin administration, TH activity was significantly reduced in the posterior pituitary. This was associated with a significant decrease in norepinephrine (NE) content. After 5 weeks, TH activity and NE content were no longer significantly different from controls. Dopamine (DA) content of the posterior pituitary was decreased progressively by melatonin administration, with a reduction of greater than 50% after 5 weeks of treatment. These data provide evidence that melatonin has a potent inhibitory effect on the regulation of the dopaminergic system of the neurointermediate lobe--an effect that appears unrelated to changes in axonal TH. PMID- 8221134 TI - Effects of intra-PVN injections of d- and l-norephedrine on feeding in rats. AB - Phenylpropanolamine (PPA) is thought to inhibit feeding by activation of alpha 1 adrenergic receptors within the paraventricular hypothalamus (PVN). Systemic injections of the PPA component enantiomers, d- and l-norephedrine (NEP), result in differential suppression of feeding (l-NEP more potent than d-NEP). Whether the norephedrine racemates induce differential anorexia subsequent to injection into the PVN is unknown. In the present study, adult male rats received intra-PVN injections of the d- and l-norephedrine enantiomers (0, 80, 160, and 240 nmol). Significantly greater anorexia was obtained for l-NEP relative to d-NEP. These results document a stereospecific effect of the norephedrine enantiomers within the PVN in inhibiting food intake and suggest that the interaction of these enantiomers with PVN alpha 1-adrenoceptors may mediate the similar difference in potency noted for systemic injections of d- and l-norephedrine. PMID- 8221135 TI - Mating-induced increases in FOS protein in preoptic area and medial amygdala of cycling female rats. AB - Genitosensory stimulation received during mating initiates neural and endocrine changes necessary for pregnancy. The present study examined sites of induction of the proto-oncogene, c-Fos, after mating in the cycling female rat to determine neural sites activated by such stimulation that might be involved in pregnancy initiation. Two groups of cycling female rats were exposed to males on the evening of proestrus and remained with the male until receiving either 14 mounts with-intromission including ejaculations (Intromissions) or a control number of mounts-without-intromission (Mounts-Only). These two mating treatments were previously shown to induce pregnancy/pseudopregnancy in 100% and 0% of the animals, respectively. Seventy-five minutes after mating, females were perfused intracardially with 2% paraformaldehyde/2.5% acrolein, and the brains were processed for FOS immunocytochemistry using standard procedures. FOS immunoreactive cells (FOS-IR) were counted in standard template quadrilaterals in the preoptic area (POA), medial amygdala (mAMYG), and the paraventricular (PVN), ventromedial (VMN), and dorsomedial (DMN) nuclei of the hypothalamus at 50 x magnification using a camera lucida. Significantly higher numbers of FOS-IR cells were seen in Intromissions females above Mounts-Only females in the POA and mAMYG, demonstrating that c-Fos expression in these areas depended upon cervical vaginal stimulation rather than on cutaneous somatosensory input received in Mounts-Only tests. The cells within the POA and mAMYG that are activated in response to the intromittive stimulus may be directly involved in pathways triggering one or several of the neuroendocrine responses to mating in the female rat. PMID- 8221136 TI - CCK-8 excites oxytocin-secreting neurons in the paraventricular nucleus in rats- possible involvement of noradrenergic pathway. AB - Systemic administration of CCK-8 increased plasma oxytocin (OXT) level in rats anesthetized with a mixture of urethane and alpha-chloralose. Extracellular recordings were made from magnocellular neurosecretory neurons in the paraventricular nucleus (PVN) of the hypothalamus in anesthetized rats to examine the effects of CCK-8 on the firing of PVN neurons. Thirteen out of 16 nonphasic neurons (putative OXT-secreting neurons) were excited by intravenous and/or intraperitoneal administration of CCK-8. By contrast, 8 out of 10 phasic cells, vasopressin(AVP)-secreting cells, were inhibited by systemic administration of CCK-8. Four out of five nonneurosecretory neurons in the PVN were excited by the administration of CCK-8. Moreover, microiontophoretically applied phentolamine blocked the excitatory responses induced by CCK-8 in nonphasic neurons. We measured extracellular noradrenaline (NA) level in the PVN, using in vivo microdialysis. Intravenous administration of CCK-8 induced NA release in the PVN. These results suggest that CCK-8 activates the excitatory afferent pathway to putative OXT-secreting neurons in the PVN which may, at least in part, be involved in the central noradrenergic projection. PMID- 8221137 TI - Visceroendocrine responses elicited by neuropeptide Y in the nucleus tractus solitarius. AB - Neuropeptide Y (NPY) has been shown to be localized in a number of CNS regions, including the nucleus tractus solitarius (NTS). In this meeting report, a brief overview is presented of recent studies from our laboratory examining the role of NPY in NTS-mediated mechanisms of cardiorespiratory and visceroendocrine regulation. Microinjections of NPY, NPY analogs, or C-terminal NPY fragments were made into the subpostremal NTS of anesthetized spontaneously breathing rats. NPY elicited pronounced dose-related depressor responses, bradycardia, and reductions in respiratory minute volume. The overall cardiorespiratory response pattern elicited by NPY was mimicked by NPY, a fragment of NPY exhibiting selective agonist properties at presynaptic Y2 receptors, whereas the Y1 receptor-selective analog, [Leu31,Pro34]NPY, and the C-terminal inactive fragment, NPY, were found to be ineffective. In an effort to further characterize intrinsic NTS mechanisms mediating the NPY-evoked response pattern, NPY microinjections were similarly made in a group of rats with bilateral glossopharyngeovagotomy (G-vagotomy) and in a group of rats decerebrated at the supracollicular level. The results showed that whereas decerebration did not appreciably affect the NTS-mediated cardiorespiratory responses elicited by NPY, G-vagotomy enhanced the NPY-evoked hypotension while at the same time abolishing the NPY-evoked bradycardia and reductions in tidal volume. Taken together, these observations with G-vagotomized animals, along with the results from microinjection studies using selective ligands for NPY receptors, suggest that NPY may modulate primary visceral afferent information via activation of Y2 receptors distributed at presynaptic sites in the subpostremal NTS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221138 TI - Olfactory bulb neurons respond to cervicovaginal distension. AB - Mitral cell layer neuronal activity in the olfactory bulb (OB) of the anesthetized rat is modulated by cervicovaginal distension. Data are reported on 22 cells that decreased and 6 that increased in response to the distension. These results provide support for the existence of a functional interaction between the reproductive tract and the olfactory system. PMID- 8221139 TI - Effect of chronic antidepressant treatment on responses to apomorphine in selectively bred rat strains. AB - The purpose of this study was to verify the dopamine-sensitizing behavioral effect of chronic antidepressant treatment in two selectively bred rat strains: the hypercholinergic Flinders Sensitive Line (FSL) and control Flinders Resistant Line (FRL). Two antidepressants, desipramine HCl (DMI) and sertraline HCl, were injected IP in separate groups of FSL and FRL rats in a dose of 16.5 mumol/kg twice daily for 16 days. Twenty-four hours after withdrawal, locomotor and hypothermic responses to 0.2 mg/kg of apomorphine, SC, were examined. Attenuation of the effect of apomorphine was observed in the open field: FRLs withdrawn from sertraline were significantly less mobile than control FRLs, and the same trend was found in FSL rats. Chronic DMI resulted in similar changes in the locomotor activity. Sertraline treatment decreased apomorphine-induced hypothermia by almost half in FSLs, whereas slight hyperthermia was induced in FRL rats instead. The present results suggest that in these selectively bred strains, a serotonergic antidepressant such as sertraline may have sensitized dopaminergic autoreceptors and/or desensitized postsynaptic receptors. Apomorphine-induced hypothermia could be mediated by serotonergic neuron function that may have been altered by chronic sertraline but not DMI treatment. PMID- 8221140 TI - Locomotion of aged rats: relationship to neurochemical but not morphological changes in nigrostriatal dopaminergic neurons. AB - Spontaneous locomotion and motor coordination was evaluated in young (5-6 month old) and aged (24-25 month old) rats. Animals were tested for spontaneous locomotor activity in Digiscan Animal Activity Monitors during the nocturnal cycle. Aged animals exhibited a significant hypoactivity compared to their young counterparts. Evaluation of the time course of activity revealed that the young animals had a cyclical pattern of activity during the 12-hour testing period with clear peaks at 2-4 hours after the initiation of testing and at 8- to 10-hour intervals thereafter. In contrast, the aged animals exhibited a blunted initial activity peak. During the remainder of the test period the aged animals activity was stable with no further peaks in activity. Compared to the young animals the aged animals also (a) remained suspended from a horizontal wire for less time, (b) were unable to descend a wooden pole covered with wire mesh in a coordinated manner, (c) fell more rapidly from a rotating rod and (d) were unable to maintain their balance on a series of wooden beams with either a square or rounded top of varying widths. Histological analysis demonstrated that there was no reduction in the number, area, or length of tyrosine hydroxylase-immunoreactive neurons within the A8, A9, or A10 region of the aged animals. Neurochemical analysis revealed that while DA and HVA levels were not decreased in the aged rats, DOPAC levels, as well as the ratios of DA/DOPAC and DA/HVA, were decreased. These results indicate that neurochemical but not morphological changes within the nigrostriatal dopaminergic system underlie the deficits in motor behavior observed in aged rats. PMID- 8221141 TI - Behavioral effects of fetal neural transplants: relevance to Huntington's disease. AB - Animal models of Huntington's disease (HD) and other neurological disorders have proven useful for examining the anatomical, neurochemical, and behavioral alterations in these diseases. Investigators have taken advantage of new excitotoxic models that appear to successfully simulate the neurobiological and behavioral characteristics of HD with remarkable homology. Selective excitotoxic compounds allow for a more precise and controlled lesion with which to examine the relationship between striatal damage and behavioral abnormalities. In addition, these models provide new approaches for developing and testing various treatments for HD. Fetal neural tissue transplanted into the excitotoxin-lesioned animal can integrate with the host brain and promote neurochemical and functional recovery. Neural grafting paradigms may be viewed as potential therapies for treating neurodegenerative diseases and as aids in deciphering the regenerative mechanisms of the central nervous system. Further research is necessary, however, to determine the negative and positive effects of neural transplantation. In addition, existing behavioral models need to be refined to allow for better evaluation of the subtle topographic changes in behavior resulting from fetal tissue transplantation. PMID- 8221142 TI - Angiotensin II(3-8) (ANG IV) hippocampal binding: potential role in the facilitation of memory. AB - The present research characterizes a newly discovered ANG II(3-8) (ANG IV) binding site localized in structures associated with memory function (hippocampus, neocortex, cerebellum), as well as other brain stem structures (thalamus, inferior olivary nucleus). This site is not the AT1 or AT2 site that binds angiotensins II (ANG II) and III (ANG III) nor does it bind the nonpeptide AT1 or AT2 receptor antagonists DuP753 and PD123177, respectively. The intracerebroventricular (ICV) infusion of ANG IV was ineffective at inducing drinking in rats as compared with equivalent doses of ANG II and III. Although not as effective as ANG II or ANG III, ICV infusion of ANG IV did provoke a pressor response at the highest dose (100 pmol/min), which appeared to be mediated by ANG II (AT1)-type receptors and not the specific AIV binding site described here. By contrast, the ICV infusion of ANG IV resulted in greater effects upon retention and retrieval of a passive avoidance task as compared with ANG II. Specifically, ANG II was not different from the ICV infusion of artificial cerebrospinal fluid, while ANG IV improved retention and retrieval of this task. PMID- 8221143 TI - Long-term habituation to spatial novelty modifies posttrial synchronized sleep in rats. AB - To assess the role of posttrial synchronized sleep in the processing of a nonassociative task, adult male Sprague-Dawley rats with chronically implanted cortical electrodes for EEG recording were exposed to a Lat-maze, and horizontal (HA; corner crossing) and vertical (VA; rearings) activities were monitored during two 10-min test trials made at a 3-h (experiment 1) or 24-h (experiment 2) interval. EEG conventional recording was taken during 3 h under baseline conditions (day 1), and following exposure to the maze (day 2), and analyzed as to the amount (a), number (n), and mean duration (d) of synchronized sleep (SS) episodes followed by wakefulness (SS-->W) or by paradoxical sleep (SS-->PS). In both experiments there was a significant intertrial decrement (long-term habituation: LTH) for horizontal activity (LTH-HA), vertical activity (LTH-VA), and emotionality (LTH-E). In experiment 1, in comparison to baseline values, the posttrial SS-->PS(a) increased, mainly for the appearance of SS-->PS episodes in the 1st h. SS-->W(a) also increased in the first h. Correlative analyses among behavioral and sleep parameters showed that SS-->PS(n) and (d) covaried positively with LTH-HA relative to the entire test, and with LTH-VA relative to the second part of the test in the third h. Negative correlations were present between SS-->PS(n) and (d), and LTH-E. In experiment 2, exposed rats showed a lower SS-->PS(n) in the first hour and an increased SS-->PS(d) in the second hour. No change was observed as to SS-->W episodes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221144 TI - Distributed changes in c-Fos and c-Jun immunoreactivity in the rat brain associated with arousal and habituation to novelty. AB - The effects of exposure to spatial novelty on expression of the immediate early gene (IEG) products c-Fos and c-Jun were mapped in the rat brain by immunohistochemistry. Adult male Sprague-Dawley rats were tested for 10 min in a Lat-maze, and corner-crossings, rearings, and fecal boli were recorded. Rats were sacrificed at different time intervals after exposure to the maze (0.5, 2, 6, or 24 h). Unexposed rats or rats repeatedly exposed for 3 days at 24 h interval served as controls. Nonperfused brains were processed for immunocytochemistry for c-Fos and c-Jun on adjacent slices using the avidin-biotin method and diaminobenzidine as chromogen. In unexposed control rats the constitutive expression of the two IEGs products was low and scattered. In contrast, rats that had been exposed for the first time to the maze (spatial novelty) showed an extensive c-Fos- and c-Jun-like immunoreactivity in the reticular formation, the caudate-putamen complex, the hippocampus (granular and pyramidal neurons), the cerebellum (granular neurons), and all layers of somatosensory cortex. The positivity was stronger in rats exposed for the first time to the box than in repeatedly exposed or unexposed control rats. A maximal IEG expression was found in animals with postexposure survival times of 2 and 6 h. IEG expression in repeatedly exposed rats was still above baseline expression of unexposed rats but still lower than that of rats having been exposed only once to the maze.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221145 TI - Striatal grafts in the ischemic striatum improve pallidal GABA release and passive avoidance. AB - Fetal striatal cells were grafted into the ischemic striatum of rats and pallidal GABA release, and behavioral improvement were investigated. Intraluminal occlusion of the middle cerebral artery (MCA) for 1 h induced ischemic infarcts in the lateral striatum and the adjacent cortex. In ischemic rats, the performance of a passive avoidance task was disturbed, and the pallidal GABA level detected by microdialysis decreased to about a half of control. After the graft, the deficit in the passive avoidance was partially alleviated and the GABA level recovered moderately and increased further by the infusion of an uptake blocker. The data indicate that fetal striatal cell grafts in the ischemic striatum partially restored both chemical and behavioral deficits. PMID- 8221146 TI - Protective effect of under-reinforcement of passive avoidance against scopolamine induced amnesia. AB - Administration of antimuscarinic drugs induces amnesia of aversively motivated behaviors. However, when relatively high intensities of footshock are used during training (over-reinforcement), animals become protected against such amnesic state. Moreover, the protective effect is established in a none-or-all fashion, i.e., within a series of increasing intensities a minute augmentation of footshock intensity is sufficient to reach the protective threshold. In the present experiment it was found that very low intensities of aversive stimulation (under-reinforcement), sufficient to produce learning, also protected animals from scopolamine-induced amnesia. These results suggest that acetylcholine is critically involved in memory consolidation of passive avoidance, but only within a limited range of training intensities. PMID- 8221147 TI - Convergence of gastric and hepatic information in brain stem neurons of the rat. AB - Convergence of gastric and hepatic information in the nucleus of the solitary tract (NTS) was investigated by single-pulse electrical stimulation of the hepatic and the gastric branches of the vagus. Facilitation induced by both stimuli occurred most often in responses of neurons in the NTS, indicating convergence of information there. In a second experiment using gastric distension and portal infusion of glucose to analyze such convergence, activation by gastric distension and suppression by portal glucose was the most prominent combination of responses to both stimuli. This confirmed the results of electrical stimulation, because distension increases the firing rate of gastric afferents and glucose infusion decreases the firing rate of hepatic afferents. Analysis of the responses to varying degrees of gastric distension revealed that some NTS neurons are activated by weak distension, but inhibited by strong distensions. PMID- 8221148 TI - Glucose-responsive neurons exist within the area postrema of the rat: in vitro study on the isolated slice preparation. AB - Responses to glucose of spontaneously active neurons were investigated by extracellular recording in the rat area postrema slice preparations (in vitro). Among 67 spontaneously active neurons, 16 neurons displayed a marked increase or decrease in discharge rate in response to increases or decreases of the glucose concentration in perfusate. These results confirm the existence of glucose responsive neurons within the area postrema suggested in prior in vivo experiments. Response to CCK or dopamine was also examined on the isolated area postrema slices. The neuron that showed a marked increase in discharge rate responding to glucose elicited a marked increase of discharge rate in response to 2.1 microM CCK, suggesting that glucose and CCK affect the same neurons. Some neurons showed a marked increase or decrease in the discharge rate in response to 20 microM dopamine, but these neurons showed neither response to CCK nor to glucose. It is likely that different neuronal networks in the area postrema contribute to control of ingestion and to initiation of nausea. PMID- 8221149 TI - Food intake, sympathetic activity, and adrenal steroids. AB - Food Intake is reciprocally related to the activity of sympathetic nerves to brown adipose tissue. This reciprocal or feedback relation is shown for hypothalamic lesions, drugs, and many peptides. These peptides also modulate intake of specific nutrients. Galanin and opioids increase fat intake, whereas enterostatin decreases fat intake. NPY increases carbohydrate intake and growth hormone releasing hormone decreases protein intake. The activity of the sympathetic nervous system is low in obesity and adrenalectomy reverses this decrease in sympathetic activity and reverses or stops the progression of obesity. One mechanism for this effect of adrenal steroids is through a transacting substance which is involved in steroid actions and the production of obesity. PMID- 8221150 TI - Macronutrient choice following food deprivation: effect of dietary fat dilution. AB - Under standard laboratory conditions rats given access to three separate macronutrient sources compose a diet yielding 31% of their total daily calories as protein, 34% as carbohydrate, and 34% as fat. This selection pattern is dramatically altered with restored access following a 48 h fast. During the first hour of refeeding, rats composed a diet that was low in protein and high in carbohydrates and fat. By the end of 24 h, no difference in selection pattern was found, though intake of all three macronutrients was higher than baseline. A separate group given access to three macronutrient sources of equal caloric density specifically increased fat intake during the period of restored access. Another group, familiarized with a concentrated fat source, was given access to a diluted fat source during refeeding. Similarly, a fourth group, familiarized with a diluted fat source during the baseline condition was given access to a concentrated fat source during refeeding. Results from these experiments suggest that prior experience with a diluted fat source promotes a significant increase in fat intake and a suppression of carbohydrate intake during initial refeeding following a 48 h fast. In a second experiment, rats that were given a choice of both fat sources preferred the concentrated source; 72% of all fat (g) and 82% of all fat calories were consumed from the concentrated fat source. These results suggest that fat intake increases following deprivation not solely due to its inherent relatively increased caloric density but also possibly due to its role in energy metabolism. PMID- 8221151 TI - Liver denervation, 5-HT3 receptor antagonist, and intake of imbalanced amino acid diet. AB - The serotonin3 receptor antagonist ICS 205-930 (ICS) may act peripherally to attenuate the anorectic response of rats given an imbalanced amino acid (IMB) diet. Rats were divided into four groups: SHAM+saline (sal); SHAM+ICS; total liver denervation (TLD) + sal; and TLD+ICS. Rats were then given a purified basal diet for 16 days. Next, the groups were injected with sal or 9 mg/kg BW of ICS at 0800 h and at 0900 h (lights out) an isoleucine IMB diet was presented. By 12 h postinjection, the food intake (FI) of TLD and SHAM rats receiving ICS was similarly higher (p < 0.02) than sal-injected counterparts whose FI was also similar; BW followed FI. By day 3, the SHAM groups had similar low FI, whereas the FI of the TLD groups was increasing. The above study was repeated with similar results. Liver innervation is not required for ICS attenuation of IMB diet-induced hypophagia. Also, while sal-injected TLD rats show a normal attenuation of consumption of the IMB diet on the first day of exposure, they subsequently consume more of the IMB diet than SHAM rats. The reason for this difference in TLD rats is not clear but may be related to metabolism of the IMB diet or possibly learning. PMID- 8221152 TI - Food consistency modulates eating volume and speed through brain histamine in rat. AB - Changes in meal parameters of rats fed with different consistency of food were examined using hard and soft pellets. Meal size and eating speed of the first meal after 1800 h increased significantly in rats fed with soft pellets compared to those fed with hard pellets. Effects of histamine depletion on meals treated with hard or soft pellets were investigated after an intraperitoneal injection of 0.11 mmol/kg alpha-fluoromethylhistidine (FMH), a specific suicide inhibitor of the histamine synthesizing decarboxylase enzyme. When rats were fed with hard pellets, FMH significantly decreased eating speed and prolonged meal duration without affecting meal size. When rats were fed with soft pellets, FMH increased meal size and duration, but not eating speed. The meal parameter of eating speed was significantly decreased and meal size and duration were increased in obese Zuckers, a hereditary histamine-depleted animal model, when compared to their lean littermates. These results indicate that proprioceptive sensation from the oral cavity may regulate meal parameters through histaminergic neurons in the brain. PMID- 8221153 TI - The effects of amygdaloid stimulation on amphetamine-elicited locomotor sensitization. AB - Systemic injection of d-amphetamine (1.0 mg/kg) resulted in a progressive increase in locomotor activity as a function of repeated daily drug administration. The magnitude of the stimulant-induced sensitization effect was enhanced by low-current electrical stimulation of the central nucleus of the amygdala during open-field testing. Amygdaloid stimulation in the absence of amphetamine treatment did not influence spontaneous locomotor activity, and there was no behavioral evidence of epileptogenesis following amygdaloid stimulation over the course of the experiment. However, with continued stimulation of the amygdala, early-stage convulsive activity was apparent in animals after approximately 40 days of testing, signifying the advancement of kindling evolution. These results suggest that the processes responsible for kindling acquisition, prior to the behavioral expression of epileptiform events, interact with the underlying substrates of amphetamine sensitization. PMID- 8221154 TI - The role of serotonergic neurons in intravenous hypertonic saline-induced secretion of vasopressin, oxytocin, and ACTH. AB - This study tested the effect of brain serotonin (5-HT) depletion on the secretion of oxytocin (OT), vasopressin (VP), and adrenocorticotropin (ACTH) due to an osmotic load. The 5-HT neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) was used to deplete brain 5-HT. The OT, VP, and ACTH osmotic sensitivity (slope of delta[OT]/delta[Osm]) and the osmotic threshold (X intercept of delta[OT]/delta[Osm]) were evaluated. Depletion of brain 5-HT decreased the OT osmotic sensitivity by > 80% (p < 0.001) without changing the OT osmotic threshold. Brain 5-HT depletion had no effect on the VP osmotic sensitivity and increased the VP osmotic threshold from 287.8 +/- 1.5 to 293.1 +/- 2.0 mOsm/kg (p < 0.05). The plasma ACTH increase due to infusion of hypertonic saline was not affected by brain 5-HT depletion. Brain 5-HT depletion significantly (p < 0.01) decreased the pituitary content of OT and VP by 38 and 32%, respectively, without changing ACTH content. These results provide evidence for a functional role of serotonergic neurons in osmoregulation of plasma and pituitary concentration of OT and VP, but not ACTH. PMID- 8221155 TI - Cardiac, neuroendocrine, and behavioral effects of central amygdaloid vasopressinergic and oxytocinergic mechanisms under stress-free conditions in rats. AB - The central nucleus of the amygdala (CEA) is considered to play a major role in the expression of behavioral, autonomic, and neuroendocrine components of the stress response. The present study was designed to examine possible modulating effects of the neuropeptides arginine-8-vasopressin (AVP) and oxytocin (OXT) on functioning of the CEA in male Wistar rats. Heart rate, neuroendocrine parameters, and behavioral activity were repeatedly measured before, during, and after local administration of several doses of AVP and OXT under stress-free resting conditions. In comparison with control artificial-CSF infusion, AVP infusion in the lowest dose (20 pg) caused in a part of the animals a long lasting decrease in heart rate, i.e., bradycardia, without affecting behavioral activity. In contrast, local infusion with high doses of AVP and OXT (2 ng) induced a transient cardioacceleration concomitant with an increase in behavioral activity. Moreover, these latter effects of AVP could effectively be blocked by pretreatment with a selective OXT receptor antagonist. These findings suggest that higher doses of AVP are effective via agonistic action on OXT receptors in the CEA. A strong correlation existed between the magnitudes of the tachycardiac response and behavioral activation. Thus, heart rate increase by OXT receptor stimulation is possibly due to somatic-autonomic coupling rather than genuine autonomic activation. Additionally, plasma corticosterone, but not epinephrine and norepinephrine, concentrations were elevated in response to AVP and OXT infusions. In conclusion, these results suggest that vasopressinergic influences on CEA function involve two receptor mechanisms possibly related to differential output systems. PMID- 8221156 TI - Differential induction of c-Fos immunoreactivity in hypothalamus and brain stem nuclei following central and peripheral administration of endotoxin. AB - Lipopolysaccharide (LPS), an endotoxin associated with gram-negative bacteria, is a potent activator of the immune system. We have tested the effects of ICV infusions of LPS (10 ng) or Ringer's solution on the induction of the proto oncogene protein c-Fos in the brain as well as plasma levels of corticosterone and splenic concentrations of norepinephrine (NE) and VIP. At 3 h post-ICV infusion of LPS, numerous labeled neurons were observed in the paraventricular nucleus (PVN) of the hypothalamus and the nucleus tractus solitarius (A2) region of the brain stem. Also, corticosterone and splenic NE and VIP levels were all elevated post-ICV LPS. Analysis of the time course for the induction of c-Fos protein in the brain following IP injections of LPS indicated that, relative to control injections, increased numbers of c-Fos-positive cells were detected in the PVN 0.5 h following IP injections (100 micrograms), peaked at 2-3 h postinjection, and then returned to control levels at later intervals. Additional dose-response data for IP LPS indicated a small increase in the number of labeled cells at a dose of 4.0 micrograms, and the number and staining intensity increased up to a dose of 100 micrograms. Corticosterone levels followed a similar pattern and were elevated at the 4.0 micrograms IP dose of LPS and increased to peak levels at 40 micrograms and higher. In contrast to ICV injections, splenic NE levels were unaltered by IP injections of LPS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221157 TI - Effects of choline-free plasma induced by choline oxidase on regional levels of choline and acetylcholine in rat brain. AB - Choline-free plasma (CFP) was induced in rats by intravenous (IV) injection of 56.0 x 10(2) units kg-1 of choline oxidase (ChO) which completely metabolized the free Ch circulating in the plasma for at least 15.0 h and caused subsequent significant decrease in the concentration of free Ch in the three brain regions examined, the striatum, hippocampus, and cortex. However, the treatment did not affect concentrations of acetylcholine (ACh) in these regions. By contrast, intraperitoneal (IP) injection of 1.0 mmol kg-1 Ch chloride resulted in a maximum concentration of free Ch in plasma in 5 min, after which tissue Ch in all regions examined increased (p < 0.001). Concomitant increases were observed in cortical and hippocampal ACh (p < 0.05) 20 min after the injection. It is thus suggested that the brain may possess compensative mechanisms to prevent the supply of free Ch from circulating to the brain during synthesis of ACh in the brain. It is also suggested that the CFP rat would be a useful and readily available animal model for future study. PMID- 8221158 TI - GABAergic neurons of the rat dorsal hippocampus express muscarinic acetylcholine receptors. AB - The expression of muscarinic acetylcholine receptors (mAChRs) in glutamic acid decarboxylase (GAD)-positive cells in the different strata of CA1, CA3, and the dentate gyrus (DG) of the dorsal hippocampus is examined by way of quantitative immunofluorescent double labeling employing M35, the monoclonal antibody raised against purified mAChR protein. Of all GAD-positive neurons, 97.5% express mAChRs. Conversely, 92.9% of the muscarinic cholinoceptive nonpyramidal neurons express GAD. These results indicate that the vast majority of the gamma aminobutyric acid (GABA)ergic neurons express mAChRs. In addition to GAD, parvalbumin (PARV) and somatostatin (SOM) are two neurochemical substances notably expressed in GABAergic neurons. In order to examine whether the entire muscarinic cholinoceptive nonpyramidal cell group can be characterized by these three GABAergic markers, a cocktail of GAD, PARV, and SOM was used in a fluorescent double-labeling experiment with M35. These results show that 97.2% of all muscarinic cholinoceptive nonpyramidal neurons can be neurochemically characterized by the content of GAD, PARV, and SOM. In conclusion, nearly all GABAergic cells express mAChRs and, conversely, virtually the entire muscarinic cholinoceptive nonpyramidal cell group belongs to the GABAergic cell population. This study, therefore, provides anatomical evidence for an extensive neuronal connectivity of the hippocampal muscarinic cholinoceptive nonpyramidal system and the inhibitory GABAergic circuitry. PMID- 8221159 TI - Thalamic- and cerebellar-projecting interpolaris neuron responses to afferent inputs. AB - Thalamic- and cerebellar-projecting interpolaris neuron responses to afferent inputs from the temporomandibular joint (TMJ) and/or the masseter muscle (Mm) were examined in rats. Of 230 neurons tested, 24 could be antidromically stimulated from the contralateral ventral posteromedial thalamic nucleus (VPM), and 27 of 91 neurons tested were stimulated from the ipsilateral posteromedial part of crus II of the cerebellar cortex. None had dual projections. The thalamic projecting neurons were recorded in the dorsomedial region of the interpolaris; most cerebellar-projecting neurons were at the medial border of the interpolaris. Ten of 24 thalamic- and 17 of 27 cerebellar-projecting neurons received nociceptive information. Afferent inputs from the TMJ and the Mm converged on 6 of 24 thalamic-projecting neurons and on 16 of 27 cerebellar-projecting neurons. In both the thalamic- and cerebellar-projecting neurons, there was no difference between the non-nociceptive and nociceptive neurons in mean antidromic latency. The results suggest that the interpolaris integrates and relays afferent inputs from deep oral structures. PMID- 8221160 TI - Circadian rhythms differ between selected mouse lines: a model to study the role of vasopressin neurons in the suprachiasmatic nuclei. AB - Mice selected for differences in nest-building behavior differ in the number of arginine-vasopressin (AVP)-immunoreactive neurons in the suprachiasmatic nuclei (SCN). Although previous efforts to link AVP-immunoreactive neurons in the SCN to clock function have failed, we show that differences in several circadian parameters are associated with differences in the number of AVP-immunoreactive neurons between the selected lines. Although an alternative interpretation is discussed, we hypothesize that these neurons may relay timing information from the circadian pacemaker in the SCN for wheel-running activity. In addition, phase response curves (PRCs) to 15-min light pulses in constant darkness also differ between the selected lines. However, these differences are not associated with the number of the AVP-immunoreactive neurons in the SCN, but are associated with the level of nest-building behavior. Compared to the Brattleboro rat, in which homozygous rats are deficient for AVP in the entire brain, our system, exhibiting a wide range of variability, has more specific utility for studying the role of the output pathways of the SCN in circadian rhythm control. PMID- 8221161 TI - Bombesin-induced hypothermia in VMH-lesioned rats. AB - Microinfusion of bombesin into the lateral ventricles (LV) of rats pretreated with insulin or acutely deprived of food has been demonstrated to reduce core body temperature. Lesions of the ventromedial hypothalamus (VMH) have been shown to produce hyperphagia, hyperinsulinemia, and to alter serum metabolic fuels. The present study examines VMH lesions as a permissive event in bombesin-induced hypothermia in rats tested at normal ambient temperature. A between-group design was used to evaluate the effect of microinjections of bombesin (1, 10, 100 ng) into the LV of rats with bilateral VMH lesions or sham lesions. Core body temperature was recorded over a 240-min period. In animals with lesions of the VMH, hypothermia was demonstrated by 30 min after injection of the 10 ng and 100 ng doses; the hypothermia persisted for 120 min. The 1 ng dose had no effect on body temperature in VMH-lesioned animals. Animals that received sham lesions of the VMH did not demonstrate a reduction in core body temperature at the maximum effective dose (100 ng) of bombesin. These results suggest that some event(s) associated with bilateral VMH lesions acts as a permissive factor in the production of bombesin-induced hypothermia at normal ambient temperature. PMID- 8221162 TI - Functional relationships between mesencephalic central gray and septum in regulating lordosis in female rats: effect of dual lesions. AB - In order to clarify the functional relationships between the lateral septum (LS) and the mesencephalic central gray (MCG) in regulating lordosis behavior, ovariectomized female rats received dual lesions in these two areas. In the first experiment, females with unilateral (right or left, R-MCGL or L-MCGL) or bilateral MCG (B-MCG) lesions were subjected to behavioral tests after the implantation of a Silastic tube containing estradiol. Lordosis was observed in only one B-MCGL female. In the R-MCGL and L-MCGL groups, most females displayed lordosis, but lordosis quotients (LQ) were significantly lower than that of the control group. These results suggest the importance of the MCG in lordosis regulation, and that there is no functional laterality in the MCG. In the second experiment, B-MCGL or R-MCGL females received bilateral LS lesions (LSL). The lordotic activity in the LSL + B-MCGL group was extremely low, being comparable to that of B-MCGL alone. On the other hand, in the LSL + R-MCGL females, the LQ was significantly higher than that of females with R-MCGL alone and was comparable to that of controls. Thus, the lateral septum plays an inhibitory role in regulating lordosis, but the influence of the lateral septum is not stronger than the facilitatory influence of the mesencephalic central gray, because the LSL could not recover the suppressive effect of the MCGL. PMID- 8221163 TI - Distribution of parvalbumin immunoreactivity in the cat brain stem. AB - We studied the distribution of parvalbumin-immunoreactive cell bodies and fibers in the cat brain stem. A high or moderate density of perikarya containing parvalbumin was observed in the periaqueductal gray, interpeduncular nucleus, nucleus of the trapezoid body, superior and inferior colliculi, and in the substantia nigra. The nucleus ruber, cuneiform nucleus, preolivary nucleus, retrorubral nucleus, paracentral division of the tegmental reticular nucleus, central and lateral tegmental fields, and the pericentral division of the dorsal tegmental nucleus had the lowest density of immunoreactive cell bodies. Moreover, a high or moderate density of parvalbumin immunoreactive processes was visualized in the nucleus ruber, substantia nigra, superior and inferior colliculi, periaqueductal gray, nucleus sagulum, cuneiform nucleus, Kolliker-Fuse nucleus, nucleus of the trapezoid body, vestibular nuclei, dorsal motor nucleus of the vagus, and in the lateral reticular nucleus. Finally, a few immunoreactive fibers were observed in the pontine gray, nucleus coeruleus, marginal nucleus of the brachium conjunctivum, nucleus of the solitary tract, inferior olive, and in the tegmental fields. PMID- 8221164 TI - Modulation of 35S-TBPS binding by GABAergic drugs in the cerebral cortex of newborn and adult rats. AB - The present study was designed to compare the allosteric modulatory effects of GABAergic drugs on 35S-t-butylbicyclophosphorothionate (35S-TBPS) binding in the cerebral cortex of newborn (5-day-old) and adult (90-day-old) rats. To examine the influence of GABA on the modulation of 35S-TBPS binding, the assays were performed in unwashed membranes (in which the concentration of GABA was dependent on the content of this neurotransmitter in vivo), and in extensively washed membranes in the presence of defined concentrations of exogenous GABA (3 microM). In unwashed membranes, the GABAA receptor antagonist, bicuculline, and the inverse agonist for benzodiazepine receptors, 6,7-dimethoxy-4-ethyl-beta carboline-3-carboxylic acid-methyl ester (DMCM) increased 35S-TBPS binding in a concentration-dependent manner in adult rats, but not in newborn rats. By contrast, in extensively washed membranes (plus 3 microM GABA) both bicuculline and DMCM were able to stimulate 35S-TBPS binding either in newborn or in adult rats. On the other hand, the inhibitory effect of diazepam on 35S-TBPS binding was observed in both unwashed and extensively washed membranes from newborn and adult rats. These results reflect the early development of the allosteric interaction between the different components of the GABAA receptor complex. In addition, the age-dependent changes in the concentration of endogenous GABA play a critical role in the modulation of 35S-TBPS binding by GABAergic drugs. PMID- 8221165 TI - An excitatory amino acid synapse in the thoracic spinal cord is involved in the pressor response to muscular contraction. AB - The increase in arterial pressure and heart rate elicited during exercise are produced by descending central command and by feedback from contracting limb muscles. Previous studies from this laboratory have demonstrated that neurons in the ventrolateral medulla (VLM) that project to the intermediolateral (IML) columns of the thoracic spinal cord are involved in the mediation of the pressor response to contraction of hind limb muscles. This study determines if these VLM neurons utilize excitatory amino acids (EAA) as the neurotransmitter at the synapse on IML neurons. The arterial pressure and heart rate responses to static muscular contraction, elicited by stimulation of the L7 and S1 ventral roots, and to electrical stimulation in the caudal hypothalamus were examined in anesthetized cats. Both muscular contraction and hypothalamic stimulation elicited significant increases in arterial pressure and heart rate. Intrathecal administration of the broad spectrum, postsynaptic EAA antagonist, kynurenic acid, greatly reduced (-77%) the pressor response to muscular contraction. A smaller (-27%) decrease in the magnitude of pressor response elicited by muscular contraction was produced by intrathecal administration of 2-amino-4 phosphonobutyric acid which acts on a presynaptic EAA receptor. Neither antagonist affected the heart rate responses associated with muscular contraction or the cardiovascular responses to hypothalamic stimulation. These results indicate that the pressor response elicited by feedback from contracting hind limb muscles is mediated through an excitatory amino acid synapse in the spinal cord. PMID- 8221166 TI - Regulation of central muscarinic receptors after cholinesterase inhibition: effect of clonidine. AB - In rats, the injection of soman (70 micrograms/kg, SC) resulted in a 90% inhibition of the cholinesterase (ChE) activities in three brain regions. The density (Bmax) for muscarinic acetylcholine receptors (mAChRs) following a single injection of soman was significantly reduced at 2 h after injection in the cortex and hindbrain. Bmax values, however, returned to baseline within 24 h. Subacute (repeated injection every 15 min) treatment with a sublethal dose of soman over 2 h also decreased the density of mAChRs. In both cases the density of mAChRs was reduced by about 15% for the cortex and 17% for the hindbrain (the midbrain was also reduced by 18% for subacute injections). Chronic administration (once daily for 7 days) of soman (20 micrograms/kg, SC) produced maximal inhibition of ChE activity but did not significantly downregulate mAChRs. Clonidine pretreatment reversed the soman-induced mAChR downregulation in cortex and hindbrain produced by acute soman administration. Thus, marked reduction in the levels of brain ChE is not the only factor involved in the production of mAChR downregulation to cholinesterase inhibitors. PMID- 8221167 TI - Long-term effects of neonatal capsaicin on C-fiber excitability and dorsal horn C input processing in the rat. AB - The effects of neonatal capsaicin treatment (50 mg/kg, SC, at day 2 of postnatal life) on C-fiber excitability and dorsal horn C-input processing were studied through recording of a C-fiber-evoked spinal flexor reflex in 55-60-day-old rats, anesthetized with urethane (1.1 g/kg, IP) and spinalized at Th9-10. Neonatal capsaicin resulted in decreased C-fiber excitability, as revealed by increased chronaxie values determined in a strength-duration paradigm. Besides, capsaicin treated rats exhibited a reduced potentiation of the C-reflex discharge in response to repetitive stimulation. The results indicate that capsaicin given to rats early in life leads to both functional disturbances of surviving C-fibers and altered temporal synaptic processing of the C-input in the spinal cord. PMID- 8221168 TI - Treatment with alpha-helical-CRF(9-41) prevents the anorectic effect of 17-beta estradiol. AB - The role of corticotropin-releasing factor (CRF) in the anorexia induced by 17 beta-estradiol (E2) has been assessed in castrated female rats that were trained to eat their daily food ration in three separate meals. Each rat was implanted with a permanent guide cannula that was aimed at the right lateral ventricle of the brain. Seven days after the brain surgery each rat was also subcutaneously implanted with an osmotic minipump containing Buserelin, a potent GnRH agonist that induces reversible castration in rats. Eight rats were used in the study, and each of them underwent four experimental treatments that consisted of a) a subcutaneous (SC) injection of oil combined with an intracerebroventricular (ICV) infusion of saline, b) a SC injection of E2 combined with an ICV infusion of saline c) a SC injection of oil combined with an ICV infusion of alpha-helical CRF(9-41), and d) a SC injection of E2 combined with an ICV injection of alpha helical CRF(9-41). Subcutaneous injections of E2 or oil were carried out the day before the ICV infusions of alpha-helical CRF(9-41) or saline. Intracerebroventricular infusions were performed 30 min before the meal for which the interaction effect of E2 and alpha-helical CRF(9-41) on food intake was determined. E2 and alpha-helical CRF(9-41) interacted on food intake; E2 brought about a 33% reduction in food intake in rats when infused with saline, whereas it was without effect when infused with alpha-helical-CRF(9-41)-treated rats. The present results provide evidence that CRF is involved in the anorectic effect of E2. PMID- 8221169 TI - [Cardio-respiratory complications of sleep apnea in obese patients]. AB - The cardio-respiratory complications of sleep apnea syndrome have been prospectively assessed in 60 patients with massive obesity and free of chronic obstructive lung disease while the associated cardiovascular diseases and the alterations of pulmonary function were taken into account. These cardio respiratory complications were observed only in patients with a number of apneas per hour of sleep greater than 20. The sleep apneas induced nocturnal hypoxemia that is frequently severe and independently correlated to the apnea index, diurnal hypoxemia and hypercapnia that are usually moderate, and presumably left ventricular hypertrophy that is not related to the development of daytime hypertension. However the nocturnal apneas were not associated with the development of an impairment of right or left ventricular function, or with the occurrence of cardiac arrhythmias or conduction disturbances. The absence of severe cardiac complications in this study may be related to the fact that the patients were relatively young and that the sleep apnea syndrome was diagnosed at an early stage of evolution. The findings of this study could help to define a more rationale approach in several therapeutic indications of sleep apnea syndrome. PMID- 8221170 TI - [Forensic medicine in the hospital]. AB - The reform of teaching hospitals which integrates all the hospital disciplines has been responsible for the creation of forensic medicine units in hospitals. Several solutions have been proposed; in Lyon this organisation has been set up within the framework of medical emergency units as that is effectively where there are the most forensic problems to solve. This integration is beneficial for our discipline: it is thus possible to set up a permanent service 24 hours a day, 7 days a week, at the disposal of the justice department and the police, and it opens the possibility of doing extra examinations like X-rays or getting the opinion of a specialist such as an ophthalmologist. But the forensic pathologist in hospital can help his colleagues to draw up a difficult certificate or give advice concerning medical law: it must be remembered that forensic medicine is not only a link between medicine and law in matters of experts' reports or certificates, it also establishes the same link between the legal world and medicine. This integration has allowed the development of clinical forensic medicine and a better understanding of the needs of the general practitioner in this area. This national experience has been broadcast in periodicals such as Journal de medecine legale et Droit medical (Journal of forensic medicine and Medical Law) and books published by Editions A. Lacassagne of which I am in charge. PMID- 8221171 TI - [Meeting with a topic: nosocomial infections]. PMID- 8221172 TI - [Nosocomial infection: clinical aspects]. AB - Nosocomial infections develop within a hospital or are produced by microorganisms acquired during hospitalization. They may involve not only patients (2 to 10 percent) but also hospital personnel. They arise from complex interactions of multiple causal factors. Patients risk factors are these that reduce the patient's capacity for resisting the injurious effects of the microorganisms and impair natural host defense mechanisms: patients with malignant disorders or immunosuppressive therapy, poor nutritional status, extensive burn wounds ... The young and the elderly are generally more susceptible to infection. Other infections are preventable. Disease causation is often multifactorial. Nosocomial urinary tract infections had the highest rate, followed by lower respiratory tract infections, surgical infections and bacteremias. The emergence of other nosocomial infections, caused by bacteria (tuberculosis), virus (HIV, hepatitis B and C virus, cytomegalovirus...), Aspergillus species or Pneumocystis carinii appears to be recent in origin and is of importance to immunocompromised hosts, other patients and hospital personnel. Nosocomial infections and their social and economic impacts require for their prevention vigorous organized hospital-wide surveillance and control programs. PMID- 8221173 TI - [Microbiology of nosocomial infections]. AB - The microorganisms causing nosocomial infections belong either to the patient himself (endogenous infections) or to hospital flora. New epidemiological tools, using molecular technologies, have renewed the body of knowledge on this flora. The commonest pathogens producing nosocomial infections include coagulase positive or negative staphylococci, enterococci, enterobacteriaceae, pseudomonas and candida. A recent trend has been shown, indicating the emergence of Gram positive cocci and fungi, when E. coli and Klebsiella tend to decrease. Bacterial resistance to antimicrobials is a growing concern, with the appearance of new resistance mechanisms such as broad-spectrum beta-lactamases in Gram negative rods, vancomycin resistance in several Gram positive species, or multiply resistance in enterococci, staphylococci, pseudomonas or mycobacteria. Some pathogens create new concerns nowadays, such as M. tuberculosis, M. avium intracellulare, Gram positive bacilli (exemplified by Corynebacterium JK), water bacteria, non-albicans Candida and other fungi. Increasing therapeutic challenge underlines the prominent role of infection control programs. PMID- 8221174 TI - [Critical study of surveillance of nosocomial infections]. AB - The institution of a continuous surveillance program is a compulsory prerequisite to the effective control of nosocomial infections. This is actually the main task of the Committee for Nosocomial Infection Control as ruled by the decree of 8 May 1988. The methodology of this surveillance relies on the continuous collection of data. It must apply to hospitals of any size and lead to practical actions, at the general or local level according to circumstances. Data concern patients and staff as well. They must be collected from different sources among which the bacteriology and hygiene laboratories play an important role. However infection records from wards constitute the cornerstones for the surveillance program: the personnel involved in the filling of the forms must be clearly identified. The scope of the surveillance and analytical methods are also discussed: a comprehensive control of the whole hospital associated with incidence measurement appears to be the best approach. Prevalence surveys are also performed. They allow a better control of continuous surveillance processes. However these processes will only be profitable if the data to be recorded have been previously specified. It is of importance to get the most comprehensive data about infectious hazards. One might rely not on global infection rates per hospital, as they have no significance, but on specific infection rates in correlation with body sites or risk factors. PMID- 8221175 TI - [Resources for the campaign against nosocomial infection in hospitals]. AB - Since 1973, the Ministry of Health has recommended to set up Infection Control Committees which became statutory in 1988. They have many responsibilities including the survey of nosocomial infection rate, the implementation of appropriate control strategies, the control of "antibiotic resistance", the training of staff, the production and the evaluation of written procedures for patients care, isolation, sterilization..., as well as they are in charge of the protection of health care workers against occupational risk of infection. Although, such committees have been appointed in more than 85% of hospitals, their true activity and the rate of nosocomial infection remain much less well known. The very low proportion of beds actually involved in a "surveillance" program, the scarcity of specific personnel, the insufficiency of institutional program of training, and the height prevalence of bacterial resistance to antibiotics are all of concern. Recently, the Ministry of Health has intensified its requirements and recommendations. However, resources allocated to nosocomial infection control remain unsatisfactory and it makes mandatory that each hospital generate a program which should be recognized as a priority. It is obvious that willingness is not sufficient and that the appointment of nosocomial infection control teams is warranted with appropriately trained personnel. However it is fair te recognize that the cost-effectiveness evaluation of various strategies remains to be done. PMID- 8221176 TI - [Socio-economic and juridical aspects of nosocomial infections]. AB - The medical and paramedical personnel shows concern about clinical and epidemiological aspects of nosocomial infections, but pays also attention to their economical and legal aspects. The acquired infections in hospital bring about economic and social consequences. It is logical to study the ratio cost/efficiency insofar as the social and human dimension of illness and suffering are taken into account and the struggle and the prevention are not only seen from their economical aspect. The legal liability of hospital or State health personnel is now well-know et the number of instituted proceedings is in continuous increase. The penal jurisdiction is the most common way. No negligence is permitted and nobody can be safe. The clarity and the dialog with the patients and their families are the best means to avoid a conflict prejudicial for everybody. PMID- 8221177 TI - [Prevention of nosocomial infections]. AB - The nosocomial infections prevention have to be subject to a concerted and multidisciplinary policy including the formation development, the specifics resources attribution, a rigorous evaluation reinforcement, and a quality research promotion. The prevention have to be done within tree nosocomial infection, transmissions classical ways: transmission by the health care workers, prevented by the hand washin up, a rigorous "hygienic care" and the isolation of infections patient, transmission by the material, involving to use valid disinfection procedure, transmission within the environment witch control have to grow within an implementation of "prosthetic" material always more used. Two other aspects have to be taken in the nosocomial infection prevention: prophylactic antibiotic use with let to prevent operative site infections, the health employee infection, bacterial or viral, have to be analysed by hygienic strategy in hospital. PMID- 8221178 TI - [Eulogy of Jean-Louis Lortat-Jacob (1908-1992)]. PMID- 8221179 TI - [Reconsideration of the role of aflatoxins in public health]. AB - The pathogenicity of aflatoxins for human has been largely demonstrated. Although aflatoxin contamination of some foodstuffs has been unquestionably established, stress should be laid upon a certain lack of information: some of the affected foodstuffs have been "forgotten" in the enumeration of contaminated products (especially soybeans); relations between mycotoxins and food balance have not yet been taken into account: for instance, the influence of some vitamins, of various compounds (carotenoids, methionine, ...), influence of deficiency in proteins (often very important in Africa) etc... aflatoxin bioavailability has not sufficiently been examined (particularly that of milk aflatoxin M1.); concerning aflatoxin evaluation in contaminated products, it should be known that their distribution is very irregular. Sampling and following analysis should thus be carried out most carefully; for these motives, some conditions and likewise regulations would can again scrutinized. PMID- 8221180 TI - [Plastic surgery and the treatment of breast cancer]. AB - For many patients conservative treatments are available at the present time in case of breast cancer. Nevertheless, radical mastectomy is to be performed in a few cases. Breast reconstruction is then able, when required by the patient, to improve her quality of life. This present study aims to discuss both the technical modalities and the indications of this type of reconstructive surgery. PMID- 8221181 TI - [Arcus juvenilis and lecithin:cholesterol acyltransferase functions. Report of a case of familial fish-eye-disease]. AB - A Fish-Eye Disease family has been recently discovered in Bordeaux, being made up 3 homozygous and 3 heterozygous patients for a recessive hereditary anomaly of LCAT. The influence of the enzyme on the plasma lipoprotein composition and its role in cholesterol efflux explain, at least for a part, the pathophysiology of the lipidic corneal clouding which is the single symptom in the homozygous patients. The comparison of the molecular biology data resulting from the analysis of the patient's LCAT gene with those which have been obtained in other FED patients as in patients with classic LCAT deficiency allows to differenciate biochemically both pathologies. It allows too the differentiation between primary and secondary (Tangier disease, apo A-I deficiency, A-I and C-III deficiency) LCAT deficiencies, which may be all associated with a Corneal arcus. The profile of the lipidic parameters most often measured in plasma (Total cholesterol, HDL cholesterol, cholesterol esterification rate, lipidogramme, apo A-I, apo A-II, LCAT mass and activity) is practically pathognomonic of this affection and consequently authorizes its differential diagnosis. In spite of the striking deficiency of HDL as of their atherogenesis preventing markers these patients do not show any sign of early cardio vascular disease. PMID- 8221182 TI - [Role of intercellular communications in hepatic differentiation and carcinogenesis]. AB - The liver epoch like other tissue epochs, occurs after that different events have induced heterogeneity in embryonic cells which result in distinct evolutionary processes. These events and those of organogenesis like "induction", are deeply dependent on cell-cell communications. Cell-cell interactions involve either soluble factors (hormones, growth factors), extracellular matrix or plasma membrane proteins responsible for cell-cell recognition and/or adhesion. All these plasma membrane signals are transduced to the nucleus and modulate the expression of groups of genes. To be functionally stable along the adult stage the liver has to maintain an ordered activity of cell renewal. This balance between proliferation and differentiation is at least in part, controlled by cell cell communications. Therefore, it is not surprising that intercellular communications are altered during hepatocarcinogenesis. They involve changes in the distribution of junctions, in the amounts of extracellular matrix components and/or growth factors which all result in modifying the differentiation/proliferation balance. Cell culture models have been used for these different studies; new in vitro systems should be set up in the near future by taking advantage of the targeted hepatocarcinogenesis in transgenic mouse. PMID- 8221183 TI - [Conservation of tap water in metallo-plastic bags: observed modifications and possible health risks]. AB - Conservation of tap water kept in soft plastic bags and stocked at different temperatures (20, 37 and 55 degrees C) and its health incidences were studied over one year by samples taken at regular intervals. Pollutants generated were studied by investigating the consequences of the choice of the plastic and of the aseptic agent used at different temperatures versus the length of the period of contact. Microbiological, chemical, and physico-chemical characteristics reported here showed the existence of an exchange between the contents and the container. The presence of two toxic substances indicates that one should be careful when using inadequate material. This is a matter to think about without further speculation. PMID- 8221184 TI - Very low frequency electromagnetic fields and health. PMID- 8221185 TI - [Video-assisted abdominal-pelvic and thoracic endoscopic surgery]. PMID- 8221186 TI - [Portrait of Jean Martin Charcot]. AB - Throughout paintings, engravings, and photography, Charcot's face and his life at the Salepetriere have become widely known. More importantly, his biographers and those who wrote about their firsthand experiences with Charcot have brought to life his authority and his penetrating eye. Charcot held his students, his patients, and all those in close contact with him under a despotic rule. His shyness and emotions hid behind a cold and impenetrable mask. Much has been written about Charcot's life at the Salpetriere. He transformed the old hospice into an institute of neurology considered internationally as a model, and its fame attracted visitors and patients from around the world. He formed a school at the Salpetriere composed of his many students. These young men gathered each Tuesday evening in the luxurious reception halls of his mansion on Boulevard Saint-Germain. There they mixed with writers, artists, and politicians who were firmly republican and anticlerical. There is only information on Charcot's early years other than the major dates of his career and a few legends. Arriving at the Salpetriere in 1862, he created the foundations of neurology over the next decade by applying the anatomoclinical method. Built on the traditions of the French anamatopathological method, his system was adapted by Charcot to incorporate the new advances in microscopy and cellular pathology. Later in his career, he also directed a scientific effort towards psychophysiologic explorations of hysteria and hypnosis, some inciting severe criticism. This judgement has been revised and in its place there remains the boldness of an innovative mind. His neurological achievement remains undisputed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221187 TI - [Influence of Jean Martin Charcot on neurology, medical education and psychology in the United States]. PMID- 8221188 TI - [Imitation and utilization behavior in major depressive states]. AB - Imitation and utilization behaviour (IB ; UB) have been described in lesions or deafferentations of the frontal lobes. Both reflect a loss of the patients' autonomy. IB and UB having been encountered in a case of melancholia, a systematic study has been undertaken in psychiatric departments. Sixty patients have been examined among whom 18 had a major depressive episode and 19 a dysthymic episode. IB or UB or both were present in every case of the first group, in 3 cases of the second one. Such data show first an unexpected focal specific neurological sign in a psychiatric disease. Second, they suggest a relationship with the frontal hypoactivity shown by PET studies in major depressive episodes. Such frontal hypoactivity should not be viewed as the cause of major depression but as a concomitant disorder. Its role in the awareness and expression of the thymic disturbance is discussed. PMID- 8221189 TI - [The campaign against tuberculosis in 1993. Imperatives and current perceptions]. AB - The continuing presence of tuberculosis in the world and its reappearance in industrialized countries again emphasizes the need to maintain and reinforce tuberculosis control. Despite the existence of effective drugs, there are still, deplorably, nearly 20 million cases of active tuberculosis throughout the world, with at least 3 million deaths annually. Almost 1.7 billion people are infected with the Koch bacillus. These figures are approximate and probably underestimate the real situation, as precise epidemiological data are not available. The rules of tuberculosis control are reviewed in this article, with distinctions drawn between low prevalence countries, where the objective is to eradicate the disease, and high prevalence countries, where it must be contained and reduced as quickly as possible the interruption of the claim of transmission of tuberculosis and the necessary collective measures are described in turn. The role of new techniques, particularly those based on molecular biology, is discussed. The importance of case finding and treatment in groups at risk in France is underlined, as are the role of BCG and the importance of protective measures, particularly in hospitals and in vulnerable subjects. The association between tuberculosis and HIV is well-known, but the HIV epidemic does not seem to be the only reason for the current resurgence of the tuberculosis problem in France in particular. PMID- 8221190 TI - [Plasticity of myocardial phenotype during cardiac hypertrophy and failure]. AB - Cardiac hypertrophy and failure frequently cause complications in some cardiovascular diseases. Both conditions are associated with important modifications of the heart's contractile and endocrine functions, induced by various changes in gene expression, which in turn are attributable to chronic hemodynamic overload. Differential expression of the myosin heavy chain family leads to a disproportionate accumulation of the alpha form relative to the beta, which in turn causes slower but more efficient myocardial contraction. This transition occurs in the rodent ventricle and human atrium. In the sarcomeric actin family, both the alpha-cardiac and alpha-skeletal isoforms are expressed in the mammalian ventricle in utero. After birth, the latter transiently accumulates in the rodent ventricle at the acute phase of an experimental overload. In humans, alpha-skeletal actin accounts for over half of total actin; this ratio remains the same during heart failure. In experimental models of hemodynamic overload, and during heart failure in humans, expression of Ca(2+)-ATPase in the sarcoplasmic reticulum is reduced. This decrease may partly account for the changes in cardiac relaxation observed in these circumstances. The atrial natriuretic factor gene in the ventricular myocardium is also activated, permitting the ventricle to participate in the regulation of its loading conditions. Several mechanical and neurohumoral factors have been proposed as triggers for this gene reprogramming. Research is currently focussed on signal transduction mechanisms, and in particular identification of the transcription factors involved. PMID- 8221191 TI - [Lacteal and placental excretion of Coxiella burnetti, agent of Q fever, in the cow. Importance and prevention]. AB - Q Fever, a true zoonosis, comes always from an animal contamination. The causes of human infection are mainly the pulmonary route and at a less degree the oral route. We describe, from a mouse model, a method of quantitative evaluation of the excretion of Coxiella in the milk. Our experiments done on 400 samples of milk of clinically infected cows, show that the mammal excretion is low and irregular. Moreover it needs 10,000 times more of Coxiella to the mouse by oral route than by the intraperitoneal route. These facts seem to minimize the importance of the digestive route in the human infection. The most important cause remains the inhalation of infected dusts. The prevention of this animal excretion goes through classical steps of sanitary prophylaxis. Moreover vaccination, antibiotherapy, or association of both reduce but do not stop the excretion in the environment. The application of all or of a part of these measures will strongly reduce the human contamination. PMID- 8221192 TI - [Eulogy of Paul Mandel (1908-1992)]. PMID- 8221194 TI - [Practical training and evaluation of the attainments of surgeons of general hospital centers]. PMID- 8221193 TI - [Qualification in general surgery. Experience of the National Commission with the first action by the Counsel of licensure of physicians]. AB - The french board of qualification in general surgery examines requests from physicians who have neither a University French diplome of surgery, nor a similar diplome of a country of the European Community. From 1989 to 1991, 184 requests were submitted to the board, coming from military surgeons (n = 18), residents of university hospitals (n = 37) or general hospitals (n = 10) or from other physicians (n = 119). These latters originate mainly from non European foreign countries, and are naturalized as French citizens afterwards. The board gave 54 favourable recommendations (29%), ranging from 100% for military surgeons, to 8% for other physicians who had not been resident. Out of these requests, 18 were applied by physicians who were appointed as staff surgeon, usually in small public hospitals. Rather to appoint physicians without any surgical qualification to artificially provide jobs it would be cheaper for the care insurance system, and more secure for the patients, to close or transform hospitals which cannot find qualified surgeons. However, it is a difficult political challenge as, in France, mayors are systematically President of the administrative board of the public hospital. PMID- 8221195 TI - [Penitentiary medicine and the Rights of Man]. AB - Penitentiary medicine combines preventive medicine and treatment which are for the most part carried out in renovated structures. Other than the usual illnesses, these services and their personal are confronted with specific pathologies due to stress, tobacco consumption, insomnia, inactivity, distress, but also hunger strikes and self-mutilation. One particular concern is AIDS and its formidable complications, notably tuberculosis. The important role played by psychiatry and psychotherapeutic support in penitentiary medicine cannot be emphasized enough. Not only are many of the inmates alcoholics or drugs users, many of them are also starved for communication. The Ahens statement (1987), a true codicil of Human Rights, developed by the International Council of Penitentiary Medical Services, formally prohibits doctors to practice any form of torture or experimental medical operations on inmates, and puts medical priority above and beyond any administrative or judiciary considerations. This message is applicable to all prisons, regardless of countries: Europe, the Americas, Africa, Pacific or other. As President of the Council, we have distributed this message throughout the world, to all the various United Nations Agencies, thus teaching Human Rights, including prisoners' rights, in a concrete fashion. PMID- 8221196 TI - The accuracy of the histological classification of lung carcinoma and its reproducibility: a study of 75 archival cases of adenosquamous carcinoma. AB - The classification of lung carcinoma into a small cell lung carcinoma (SCLC) and non-small cell lung carcinoma (NSCLC), is highly reproducible. There are few studies on the reproducibility of subtyping NSCLC, with anecdotal reports on the subtype of adenosquamous carcinoma. We undertook a study of 75 cases archived as adenosquamous carcinoma. All cases were accepted as NSCLC on independent review by three pathologists utilizing the 1982 World Health Organization (WHO) classification criteria. The acceptance rate of adenosquamous carcinoma by the three pathologists was 65%, 28%, and 65%. Cases not accepted as adenosquamous fell into the different subtypes of NSCLC, with a concordance rate between each pair of pathologists of 49%, 61% and 43%, indicating poor agreement between pathologists. The study confirms the high reproducibility of the classification into SCLC and NSCLC; it shows poor concordance for all subtypes of NSCLC with poor reproducibility of diagnosis of adenosquamous carcinoma as specified by the 1982 WHO classification. Physicians dealing with lung carcinoma should be aware of this limitation of classification. PMID- 8221197 TI - Alterations in erythrocyte glutathione metabolism associated with cervical dysplasias and carcinoma in situ. AB - The study was designed to test the hypothesis whether cervical dysplasias of the more severe grades are associated with elevated erythrocyte glutathione levels. Subjects were women who obtained Pap tests and were subsequently found (1) not to have any cervical lesions or (2) to have colposcopically visualized, biopsy confirmed cervical abnormalities histopathologically diagnosed as mild, moderate, severe dysplasias, or carcinoma in situ (CIS). The erythrocyte levels of reduced glutathione (GSH), oxidized glutathione (GSSG), glutathione reductase (GR), glucose-6-phosphate dehydrogenase (G6PD), and 6-phosphogluconate dehydrogenase (6PGD) were analyzed from coded peripheral venous blood samples. GSH and GR concentrations increased with increasing severity of dysplasia. Exploratory data analysis and multiple pairwise comparisons suggested comparable levels of the glutathione-related variables between these histopathological pairs: (1) mild and moderate dysplasias or (2) severe dysplasia and CIS. We suggest that the changes in erythrocyte glutathione-related indices in conjunction with histopathological diagnosis may have the potential to distinguish between low- and high-grade cervical dysplastic lesions. PMID- 8221198 TI - Inhibition of murine sarcoma cell adherence to polystyrene substrata by bacillus Calmette-Guerin: evidence for fibronectin-mediated direct antitumor activity of BCG. AB - Bacillus Calmette-Guerin (BCG) inhibited adherence of S180 mouse sarcoma cells and WI38 human diploid fibroblasts to the polystyrene substratum of 24-well cluster dishes in a dose-dependent manner. This property was retained by washed or heat-killed bacilli, but not by the vaccine filtrate or by the spent bacterial culture medium. Adhesion of bacilli to nonadherent S180 cells was demonstrated by light and scanning electron microscopy, but was not seen after trypsinization of adherent cells, indicating that bacilli bind to cell-surface adhesins. Preincubation of bacilli with human fibronectin abolished their ability to inhibit S180 adherence, suggesting that the phenomenon may be mediated by interaction of bacilli with cell-surface fibronectin. Fibronectin pretreatment of the bacteria also decreased their inhibition of S180 tumor growth in vivo, indicating that this mechanism may be at least partly responsible for BCG vaccine's observed antineoplastic activity. PMID- 8221199 TI - A phase II study of merbarone in patients with adenocarcinoma of the pancreas. AB - Merbarone, a nonsedating derivative of thiobarbituric acid that has demonstrated antineoplastic activity against a variety of murine tumors, was evaluated in a phase II trial in patients with advanced, measurable adenocarcinoma of the pancreas. Seventeen patients were treated at a starting dose of 1000 mg/m2/day for 5 days by continuous intravenous infusion; the dose was escalated in accordance with the toxicity experienced, and no dosage reductions owing to toxicity were required. No complete or partial responses were observed, and only one minor response was documented, suggesting that merbarone is ineffective against pancreatic cancer at the doses and schedule in which it was administered in this trial. PMID- 8221200 TI - Advances in the diagnosis and treatment of testicular cancer. PMID- 8221201 TI - Gene therapy of cancer. AB - Retroviral-mediated gene transfer has permitted the development of clinical protocols for the study and treatment of cancer. These protocols can be divided into gene-labeling and gene therapy proposals. Labeling studies include the tracking of tumor infiltrating lymphocytes (TIL) following the administration of those cells, and the detection, at the time of relapse, of tumor cells from transplanted autologous bone marrow. Most gene therapy protocols are designed to induce an immune attack against the tumor by inserting genes into tumor cells themselves. Although uncertainty about the safety of the procedure still exists, gene therapy of cancer holds much promise as an effective treatment modality. PMID- 8221202 TI - Nursing salaries and practice patterns: a nationwide evaluation by the Oncology Nursing Society. AB - This article summarizes the results of a national survey of salary, staffing, and professional practice patterns in oncology nursing conducted by the Oncology Nursing Society. The respondents represent a wide geographic distribution, and data were analyzed in the aggregate, as well as by American Hospital Association (AHA) regions, to allow for predictions of regional as well as national trends. This article reports on national and regional trends in salaries for entry-level and experienced practitioners; salary patterns for agency and float pool nurses; and the methods for awarding salary increases to nursing personnel in the past year. National trends in a variety of personnel practices affecting oncology nursing (e.g., shifts, self-scheduling, tuition reimbursement, and certification) are also presented. The data in this study come from a broad-based and geographically well-distributed sample that allows for meaningful national comparisons. PMID- 8221203 TI - Screening mammography in primary care settings: implications for cost, access, and quality: background paper. PMID- 8221204 TI - Impact of cancer on couples. PMID- 8221205 TI - Recombinant antibody technology: its advent and advances. PMID- 8221206 TI - Interleukin-6. PMID- 8221207 TI - Interleukin-8: a review. PMID- 8221209 TI - Regressive penalties for NIH scientists: Alice and the White Knight's box. PMID- 8221208 TI - Interleukin-10 and cancer. PMID- 8221210 TI - "Screening mammography in primary care settings"--Wagner. PMID- 8221211 TI - Three do's and three don'ts for expert witnesses. PMID- 8221212 TI - A call for forensically relevant research. PMID- 8221213 TI - Maltreatment and the school-aged child: school performance consequences. AB - Studies of the impact of abuse or neglect on children have focused largely on maltreated infants, toddlers, or preschool children. In this study a total of 139 school-age and adolescent children participated in a multi-model, multi-source assessment; 22 of the children had been physically abused, 47 had been neglected, and the remainder served as comparison subjects. Parent and child interviews, teacher ratings, and data from school records were used to comprehensively assess children's school performance; social and emotional development in school, at home, in the community, and with peers; and adaptive behavior in areas such as motor skills, personal care skills, and community orientation. With the effects of socioeconomic status covaried out, results showed that the abused children displayed pervasive and severe academic and socioemotional problems. Neglected children differed little from children who were neither abused nor neglected on measures of socioemotional development, but they displayed severe academic delays. Both groups of maltreated children showed unexpected strengths on measures of adaptive behavior. PMID- 8221214 TI - How teachers define and respond to child abuse: the distinction between theoretical and reportable cases. AB - Teachers are considered to be among those who are well-placed for the detection and prevention of child abuse. Yet little attention has been given to the definitions teachers use in deciding which cases to process. The paper draws on a larger study of Ontario women teachers. Information from 264 elementary teachers and 47 principals was collected in three phases: exploratory interviews, a survey, and focused interviews. A key part of the survey involved 10 vignettes designed to determine how teachers define abuse, how much experience they have had in dealing with such situations, and what action they took in each case. The findings demonstrated that teachers included a broad range of behaviors in their own "theoretical" definitions, but preferred informal intervention over formal reporting. Although there is no statistical relationship between definitions and reporting, it seems clear that the decision to report involves the interplay of definitions, institutional response, and teachers' experiences with a range of reactions and personal trials. Reporting is also complicated by teachers' disciplinary role, by their concerns for establishing reasonable grounds, and by the perception that some cases can be handled more effectively by the school, without the intervention of Child Protection Services (CPS). PMID- 8221215 TI - Physicians and medical neglect: variables that affect reporting. AB - Variables that affect the propensity to report medical neglect were surveyed among 52 physicians on the staff of a pediatric hospital. Death was one of the 52 potential consequences of neglect listed for 7 of the 46 diseases analyzed. The time allotted before neglect would be considered varied with the stage and seriousness of the disease. Caretakers were less likely to be considered neglectful if the symptoms of the disease were minor, not obvious, or required technical sophistication for recognition. The number of acceptable reasons for missing appointments varied. The natural course of the disease process, efficacy and safety of treatment, parents' religion, intellectual level, and economic situation affected neglect reporting propensity. Definitions of medical neglect for common diseases must be standardized. Training of lay caretakers of ill children and communication of clear and reasonable expectations are necessary to help prevent medical neglect. PMID- 8221216 TI - Children's testimony and their perceptions of stress in and out of the courtroom. AB - Modifications of the courtroom environment have been proposed to reduce stress and enhance truth-telling of child witnesses. The present study examines the premise that courtroom environment affects the quality of children's evidence and children's perceptions of their own stress. Thirty-four 8- to 10-year-olds participated in an activity and 2 weeks later, their memory for the activity was tested. Half the children were questioned in a mock courtroom in the law school of a major university, and half at their school, both by the same interviewer. Children questioned at court showed impaired memory performance when compared with agemates questioned at school. They also rated certain court-related experiences as more stressful than peers interviewed at school. Furthermore, children's perceptions of courtroom stress were negatively correlated with completeness of accurate free recall, suggesting a relation between court-related stress and eyewitness memory worthy of further study. PMID- 8221217 TI - The combined effects of physical, sexual, and emotional abuse during childhood: long-term health consequences for women. AB - The long-term health effects of physical, sexual, and emotional abuse during childhood were studied in a sample of 668 middle class females in a gynecologic practice who responded to a self-administered, anonymous questionnaire covering demographic information, family history, physical and psychological health, as well as stressful events and abusive experiences as a child. Half (53%) of the sample reported childhood abuse, with 28.9% recounting exposure to one type of abuse, 18.7% to two types of abuse, and 5.4% to all three types of abuse. In comparison to women not abused during childhood, the abused reported significantly more hospitalizations for illnesses, a greater number of physical and psychological problems, and lower ratings of their overall health. The greater the number of childhood abuses, the poorer one's adult health and the more likely one was to have experienced abuse as an adult. Thus, in addition to the deleterious psychological consequences of abuse described in the literature, physical health also appears to be adversely affected in women abused as children. PMID- 8221218 TI - Differential understanding of sexual abuse prevention concepts among preschoolers. AB - Most evaluations of sexual abuse prevention programs employ composite knowledge scores that may mask whether individual skills are differentially understood. The few studies examining separate skills have provided no statistical justification for doing so. This study attempted to validate empirically distinct prevention skills and whether children comprehend these skills in the order taught by prevention programs: RECOGNITION (of good and bad touch), SAY-NO (refuse the perpetrator), GO (leave the situation), TELL-WHO (find an adult), and TELL-WHAT (accurately disclose the abuse). Subjects were 117 preschoolers who were individually administered the "What If Situations Test" (WIST). Analyses confirmed that the WIST was composed of six skills. The hypothesized sequence of skills produced a high coefficient of reproducibility, but an even higher coefficient was obtained when three common sequence reversals were added to the existing logical sequences. We discuss explanations for these cognitive sequence reversals and applications of this information for teaching preschoolers prevention concepts. PMID- 8221219 TI - Preschoolers' cognitions of authority, and its relationship to sexual abuse education. AB - This study investigated preschoolers' perceptions of authority in sexual and benign situations and how these might influence acquisition of concepts presented in a sexual abuse prevention program. Participants were 117 children, ages 36-84 months, at four day-care centers. Children were randomly assigned to either an experimental or delayed treatment control group. Measures of authority and prevention skills were administered prior to and following the first administration of the curriculum. Preschoolers reasoned at a higher level of authority in sexually abusive encounters as compared to benign ones. For experimental children, this capability increased after participating in the prevention program. At pretest, sexual abuse authority scores predicted scores on one of two prevention skills; pretest authority scores did not predict either posttest prevention skill. Results are discussed in relation to children's understanding of moral versus social-conventional rules, and future directions for research are indicated. PMID- 8221220 TI - Normal versus abnormal genital findings in children: how well do examiners agree? AB - In order to determine how well medical examiners agree on the significance of certain anogenital findings in children, preselected colposcopic photographs of the anogenital area of 16 patients were shown to 170 medical examiners (82% pediatricians) who were blinded as to the history on each patient. Findings rated most frequently as being suggestive or indicative of penetrating injury included immediate anal dilatation with no stool present (85%), hymenal transection (84%), marked narrowing of the hymenal rim with notching (81%), and a posterior fourchette scar (75%). The agreement between the participants and the experts on the abnormal cases (mean 81%) was significantly higher than on the normal cases (mean 71%, p < .001) and on genital findings (78%) than on anal findings (63%, p = .000). Higher experience level (more cases seen per month) was associated with significantly higher agreement between the participants and the experts on five of eight normal cases and two of four abnormal cases. Use of a colposcope was also associated with higher overall agreement with the experts (74% vs. 44%, p < .0001). PMID- 8221221 TI - Dissociative experiences and disorders among women who identify themselves as sexual abuse survivors. AB - The purpose of the study was to identify dissociative experiences and disorders among women who are survivors of sexual abuse. Fifty-one women from two different centers who identified themselves as abuse survivors participated in the research interviews. The Dissociative Experiences Scale (DES) and the Dissociative Disorders Interview Schedule (DDIS) were used to collect data. The results indicated that 88.2% of the 51 women had a dissociative disorder of some type. Twenty-eight (54.9%) of the women had a DDIS diagnosis of multiple personality disorder (MPD). The women had high DES scores, a high number of secondary features of MPD, high rates of depression, borderline personality, substance abuse, somatic symptoms, Schneiderian symptoms, ESP/supernatural experiences, suicide attempts, and conversion symptoms. The vast majority of sexual abuse survivors in this sample have extensive dissociative symptomatology and related features. Therapists working with adult sexual abuse survivors should be knowledgeable about dissociation, should thoroughly assess a client's dissociative abilities, and should incorporate these findings into the therapy process. PMID- 8221222 TI - Assisting psychosocially distressed mothers-to-be: primary prevention of child abuse. PMID- 8221223 TI - Failure to prevent physical child abuse despite detection of risk factors at birth and social work follow-up. PMID- 8221224 TI - Polarity analysis in child abuse case management. PMID- 8221225 TI - Protecting seriously mistreated children: time delays in a court sample. PMID- 8221226 TI - [Fetal thymus and thymuline stimulate in vitro proliferation of gonocytes in the fetal testis in rats]. AB - Testes from 13.5-day-old rat foetuses were cultured in vitro for 1 or 2 days: 1) in synthetic medium; 2) in thymulin- or corticosterone supplemented medium; 3) in co-culture with fragments of foetal thymus; 4) in co-culture with fragments of foetal thymus in corticosterone-supplemented medium. The numbers of germ cells were about 2-fold greater in testes cultured with fragments of thymus, or in thymulin-supplemented medium than in all the other experimental conditions. These results indicated that thymulin can stimulate foetal gonocytes proliferation and that corticosterone inhibits thymulin-stimulated gonocytes proliferation. The data suggest a functional relationship between corticosterone, thymulin and the foetal gonocytes in vivo. PMID- 8221227 TI - [Immunoreactivity to glial fibrillary acidic protein (GFAP) in the retina of the lamprey Lampetra fluviatilis]. AB - This study is the first demonstration of glial fibrillary acidic protein (GFAP) immunoreactivity in the retina of the lamprey Lampetra fluviatilis. This immunoreactivity is expressed on one hand, in radial processes and somata which belong to Muller cells and, on the other hand, in horizontal fibers in the intermediate plexus between horizontal cells. The tracing of these fibers to Muller cells or horizontal cells is discussed. PMID- 8221228 TI - Imaging by fluorescence videomicroscopy of individual single stranded DNA molecules in solution. AB - We demonstrate that the observation by fluorescence video-microscopy of individual single stranded DNA molecules is now possible owing to the use of an asymmetric cyanine (YOYO-1) belonging to a new class of fluorescent dyes complexing the DNA. The fluorescence of labelled DNA persists in denaturing conditions showing that the dye does not attach simply to the possible secondary structures. Furthermore it is stable under electrophoresis and enables the imaging of stretched molecules. PMID- 8221229 TI - [A second gene involved in the formation of disulfide bonds in proteins localized in Escherichia coli periplasmic space]. AB - A novel mutant of Escherichia coli was isolated whose phenotype is similar to those of dsbA strains. For instance, it is unable to express pH 2.5 acid phosphatase, glucose-1-phosphatase and alkaline phosphatase in the periplasmic space. The mutation lies at min 26.2 of the linkage map, and does not affect expression of DsbA. Addition of oxidized glutathione to the growth medium restores the wild-type phenotype in the mutant while this is not the case in a dsbA strain. The product of this new gene dsbX is thus actually involved in the formation of disulfide bridges in the periplasmic space but can only operate if DsbA is functional. PMID- 8221230 TI - Exclusion of genes coding for proteins of the cytoskeleton and the extracellular matrix in familial hypertrophic cardiomyopathy using a candidate gene approach. AB - Familial hypertrophic cardiomyopathy (FHC), a primary cardiac pathology, is a genetically heterogeneous disease, with autosomal dominant inheritance. The first gene identified as responsible for FHC codes for beta-myosin heavy chain (beta MHC). To find a second locus, a candidate gene approach was applied on two families for which the beta-MHC locus was excluded. Selection of candidate genes is based on the observation of tissular and cellular disorganisation in FHC, and included genes coding for proteins involved in human myocardium architecture: the extracellular matrix components and cytoskeleton proteins. Chromosomal areas containing the candidate genes were examined by linkage analysis with microsatellite markers. The genes coding for different types of collagens, laminins, fibronectin, fibrillins, desmin, titin, alpha-actinin, vinculin, cardiac and skeletal alpha-actins, ankyrin and spectrin were excluded as responsible for FHC. PMID- 8221231 TI - Localization of 15 cosmids on human chromosome 22 by fluorescence in situ hybridization. AB - Fluorescence in situ hybridization has been used in a cytogenetic analysis to map 15 cosmids on human chromosome 22. Thirteen cosmids were localized on the long arm of chromosome 22 (22q) while two other probes displayed a hybridization signal on 22p and the short arm of the acrocentric chromosomes of groups D and G. The regional assignment of these new chromosome markers will improve the mapping of chromosome 22; they can be used to detect numerical and structural aberrations of this chromosome involved in numerous pathologies. PMID- 8221232 TI - Inhibition of interleukin-2 receptor alpha-subunit gene expression by oligonucleotide-directed triple helix formation. AB - Oligonucleotide-directed triple helix formation represents a promising approach to block gene expression at the transcriptional level. We have previously shown [10] that a triple-helix-forming oligonucleotide was able to inhibit promoter function of reporter constructs in live cells, provided that the oligonucleotide was covalently linked to an intercalating agent which stabilizes triple-helical complexes. In order to demonstrate that this inhibitory effect was due to triple helix formation, we have mutated the oligonucleotide target site in the promoter of the interleukin-2 receptor alpha-chain gene. The mutated version of the promoter does not bind, and is not inhibited by the oligonucleotide, demonstrating that the observed inhibition of the wild-type promoter is indeed due to triple helix formation within cells. PMID- 8221233 TI - Two factors, IRF1 and KBF1/NF-kappa B, cooperate during induction of MHC class I gene expression by interferon alpha beta or Newcastle disease virus. AB - The expression of class I genes of the Major Histocompatibility Complex is stimulated by IFN. The promoter of these genes contains an interferon response sequence (IRS) which overlaps the major enhancer. These elements are recognized by several protein factors, including IRF-1, which binds the IRS, and KBF1/NF kappa B, which binds the enhancer. We demonstrate here that infection by Newcastle Disease Virus (NDV) results in an increased expression of class I genes, by a mechanism partially different from that of IFN, but that in both cases the cooperative action of IRF1 and KBF1/NF-kappa B is required. In F9 embryonal carcinoma cells, where KBF1/NF-kappa B activity cannot be detected, both types of stimuli are ineffective. PMID- 8221234 TI - [The corticorubral projection in rats: topographic distribution of fibers arising from areas of the sensorimotor cortex functionally identified by microstimulation]. AB - The combination of microstimulation and anterograde tracing techniques reveals that different regions of the sensorimotor cortex project to different sectors of the red nucleus (NR). V-FEF projects to the dorsolateral sector, JLT to the dorsal sector, RFL and CFL to the dorsomedial and medial sectors, respectively, and HL to the ventral and ventrolateral sectors. Moreover, the NR receives fibers from the 2 somatosensory cortical areas examined in this study. The vibrissae area projects to the dorsolateral sector and the hand area to the dorsomedial sector. PMID- 8221235 TI - [Early effects of neutron-gamma or gamma irradiation on the group toxicity induced by (+) amphetamine in mice]. AB - To better understand the mechanism of action of gamma and neutron radiation on the dopaminergic system, the influence of the two irradiation modalities on the group toxicity of (+) amphetamine was studied in mice. Neutron-gamma irradiation (3.6-4.95 Gy) leads to an early toxicity reduction, while gamma-exposure (7-12 Gy) induces an increase in toxicity. This suggests that these two types of radiation induce different early effects on central dopaminergic system. Possible mechanisms are discussed. PMID- 8221236 TI - Maturation and integration of purified foetal neurons transplanted into the adult brain. AB - Fetal neural transplants presently developed as a therapeutic strategy for neurodegenerative diseases include both the neurons of interest and their cellular environment. These glial and vascular cells may be detrimental by, for instance, expressing foreign MHC antigens. This study was undertaken to determine whether purified neurons would survive transplantation into an adult host brain. Embryonic rat spinal neurons were purified by panning and transplanted into adult hosts' brain. During the first three weeks post-transplantation the grafts contained essentially packed immature neurons. Later transplants contained large, multipolar neurons, demonstrating the ability of transplanted neurons to mature in the adult environment. The adult host appears actively involved in the integration of such a transplant by complementing it with microglial and vascular cells. PMID- 8221237 TI - [Implication of melatonin in the control of the interdigestive ileocecal-colic electromyographic profile in rats]. AB - In the interdigestive period, intestinal motility displays myoelectrical complexes occurring cyclically at a constant rhythm. Since it is established that the pineal gland plays an important role in the control of circadian rhythms via its melatonin secretion, we decided to study the effects of pinealectomy and melatonin on the organization of the ileo-ceco-colic electromyogram of the rat. Pinealectomy resulted in the disappearance of ileal myoelectrical complexes and, concurrently, an excitomotor effect on the cecocolic tract. In pinealectomized rats a single intravenous injection of melatonin during lighting period induced an immediate recovery of the ileal myoelectrical complexes and a progressive return to the control electromyographic pattern of the ceco-colic tract. The melatonin action was definitive. PMID- 8221238 TI - [Effect of the mutation muscular dysgenesis on the mitochondrial metabolism of fibroblasts in vitro]. AB - Muscular dysgenesis is a lethal mutation characterized by an absence of striated muscle contraction. This abnormality of function caused by a defect in excitation contraction coupling is due to the lack of the alpha 1 subunit from the dihydropyridine receptor in the transverse tubule membrane. A phenotypical reversion is obtained in vitro by addition of normal cells, as fibroblasts, in dysgenic muscle culture. Perisynaptic fibroblasts from denervated muscle have been shown to synthetize molecules which are usually concentrated at the synapses suggesting that signal arising after denervation may influence fibroblasts synthesis ability. We show here that the energetic metabolism level from dysgenic fibroblast is lower than that from normal fibroblasts. The synthesis or the release of muscular signals that are involved in fibroblast differentiation could be disturbed in mdg/mdg myotubes. PMID- 8221239 TI - Metal identification in human anti-hemophilia A factor (factor VIII). AB - Anti-hemophilia A factor (FVIII) consists in different heterodimers of heavy and light chains associated by a metallic ion. The integrity of this complex is indispensable for procoagulant activity. Atomic absorption spectrometry with Zeeman effect has been applied to determine the nature of this metal. For this purpose, the different active forms of FVIII were separated by FPLC and characterized by SDS-PAGE. Two peaks were observed, the first corresponding to different FVIII complexes of high molecular mass (ranging from 210-80 kDa to 110 80 kDa) and the second to the heavy-light chain dimer of 90-80 kDa. In all these active fractions, copper atom was identified and a proportionality was measured between the metal concentration and the coagulant activity. Furthermore, the determination of copper and FVIII concentrations indicated that only one copper atom is implicated in the 90-80 kDa association. PMID- 8221240 TI - [The production and significance of interleukin-4 in patients with idiopathic nephrotic syndrome]. AB - Samples of serum and PBMC were collected at the same time from 42 pediatric patients with idiopathic nephrotic syndrome (INS) and 28 age-matched healthy individuals. The level of IgE in serum was detected and IL4 in supernatant of PHA activated PBMC assayed by sandwich ELISA. The level of IL4 and IgE in healthy individuals was 400-500 pg/ml and 50-100 U/ml respectively, and that of IL4 and IgE in the INS patients was on the average 1,080-4,000 pg/ml and 380-1,000 U/ml. Positive correlations were noted between the levels of IL4 and IgE, IL4, and quantity of proteinuria in 24 hours in the INS patients. It is suggested that the elevated IgE in the INS patients was induced by the elevated IL4, and that the imbalanced function of T cells and IL4 was contributable to INS. PMID- 8221241 TI - [The association of Graves' disease with serum soluble interleukin 2 receptor]. AB - The serum soluble interleukin 2 receptor (sIL2R) was measured in 38 first visited patients with Graves' disease and 29 normal controls. The serum sIL2R in 17 patients with Graves' disease was determined after treatment with antithyroid drugs (propylthiouracil) for a short period (1.2 +/- 0.5 months). The serum sIL2R was measured by sandwich enzyme linked immunosorbent assay. The sIL2R was significantly higher in patients before (3.04 +/- 0.19 U/ml) and after treatment (2.56 +/- 0.41 U/ml) than in normal controls (2.20 +/- 0.27 U/ml, P < 0.01). The mean value of serum sIL2R in 17 patients after treatment (2.56 +/- 0.41 U/ml) was substantially decreased as compared with that before treatment (2.99 +/- 0.14 U/ml, P < 0.01). The serum level of sIL2R in pretreatment patients was correlated significantly with T3(r = 0.5032, P < 0.05), but was not obviously related to T4 or rT3. These findings suggest that the human lymphocytes in patients with Graves' disease were activated in vivo and that sIL2R may be an useful immunological indicator of disease activity. PMID- 8221242 TI - [The precursors and effectors of human lymphokine-activated killer (LAK) cells]. AB - Natural killer cells were enriched in the second fraction (F2) of discontinuous Percoll density gradients from human PBL after centrifugation, whereas T lymphocytes were enriched in F4. The cytotoxicity against K562 by F2 LAK (F2 cells activated by rIL2) was higher than that of F2 cells. F2 LAK also acquired the ability to lyse NK-resistant tumor cell lines and noncultured lung cancer cells (6 out of 9 cases). All these activities of F2 LAK cells were significantly higher than those of F4 LAK cells. Besides, F2 LAK exhibited higher level of lysis against noncultured lung cancer cells than did LAK derived from PBL. Enzyme labeled immunocytochemistry staining showed that the number of CD16+ and CD8+ cells in F2 was much more than that in F4. So the higher lytic ability of F2 LAK might come from two kinds of precursors: CD16+ typical NK cells and MHC unrestricted CD8+, NK-like' cytotoxic T lymphocytes. The number of CD4+ cells in F4 was much more than that in F2. They shouldn't be the precursors of LAK cells, otherwise the lytic ability of F4 LAK should be higher than that of F2 LAK. After being cultured with rIL2 for 3 days, F2 cells exhibited proliferative response. But the percentage of CD16+ and CD8+ cells in F2 LAK did not change much, and remained much higher than that in F4 LAK. So the effector cells of short-term cultured LAK might be the activated CD16+ NK cells and CD8+, NK-like' cytotoxic T lymphocytes as well.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221243 TI - [Types of anticellular structure antisera in scleroderma patients]. AB - The sera of 338 scleroderma patients were cytologically examined with indirect immunofluorescence method. The sera were classified according to the cellular structure, to which they were against respectively. Among these sera, six main types of cellular structure antiserum were found: antikinetochore, antinucleolus, antichromatin, antiheterochromatin, antichromosome and anticentrosome. Some of these main types can further be subtyped with respect to the substructure or the chemical components (antigens) of a particular cellular structure. These types of antiserum may possibly be correlated with the clinical signs of scleroderma patients. For exploring the possibility to use for diagnosis. PMID- 8221244 TI - [Synthetic oligopeptide of hepatitis D virus antigen and its clinical application]. AB - A 27-peptide fragment derived from natural HDAg sequence was selected, identified and synthesized. It was used to develop an EIA method for detection of anti-HD. The 27-peptide possessed the similar antigenicity with the corresponding fragment of natural HDAg, and could also compete with natural HDAg for serum anti-HD. High specificity and no cross-reaction were found among the peptide and normal human sera, normal mice sera and sera positive with HAV, HBV or HCV antibody alone. 36 serum samples were identified previously, with Abbott kits with a coincident rate of 97.2%. Anti-HD was also detected in serum samples of some blood donors and patients with liver diseases and HBV infection from 1990-1992. One (0.33%) of 300 blood donors was positive (two times higher than normal ALT level in serum). 62 patients with hepatitis A and 58 patients with non-hepatitis B were anti-HD negative. 100 (11.64%) of 859 patients with HBV infection were positive: ASC 13/410 (3.17%), AH 7/63 (10.29%), CPH 1/9 (11.11%), CAH 22/121 (18.18%), SH 15/75 (20.00%), LC 23/78 (29.49%), PHC 19/89 (19.39%). The results are consistent with our previous reports. PMID- 8221245 TI - [Protective action of taurine on ischemia-reperfusion liver injury in rats and its mechanism]. AB - The protective effect of taurine on ischemia-reperfusion damages of rat perfused liver and its possible mechanism were investigated. Reflow with 20 mmol/L taurine markedly ameliorated reperfusion injury of post-ischemic liver as evidenced by reduction of lipid peroxidation (LPO), mitochondrial calcium overload in hepatocytes, as well as leakage of intracellular LDH and protein. Furthermore, taurine could significantly suppress in vitro LPO formation of hepatocyte plasma membrane induced by generating system of free radicals, inhibit calcium uptake and promote calcium release by mitochondria in addition to a membrane stabilizing action on rat isolated hepatic lysosome. The results demonstrated that taurine possesses significant protective efficacy on ischemia-reperfusion liver damage, and that the mechanism of its anti-hepatic injury may be mainly related to inhibiting LPO formation, regulating cellular calcium homeostasis and stabilizing biological membrane. PMID- 8221246 TI - [Electrochemical analysis in differential diagnosis of benign and malignant oral maxillofacial tumors]. AB - A bioelectrochemical-sensor device was designed according to the principles of galvanic cell reaction. Modified bioelectrochemically, the device was used to measure the current of samples from benign and malignant tumor tissues. Statistical analysis and clinical test showed that the current values (426 microA) of the malignant tissue were higher than those (216 microA) of the benign tissue. The sensitivity and specificity were 95% and 90% respectively. This method provided important information about benignancy or malignancy of the tumor and its involvement and metastasis. PMID- 8221247 TI - [Lectins in gastric carcinoma and precancerous lesions]. AB - Three lectins (PNA, PHA, dBA) were used to study gastric carcinoma (66 cases), lymph node metastatic tumors (32), normal gastric mucosa (10), dysplasia (30), and intestinal metaplasia with gastritis (43). The intestinal metaplasia with quite abundant PHA and DBA receptors was closely related to gastric carcinoma. The positive rate of PHA receptors increased progressively in order of mild dysplasia, moderate dysplasia, severe dysplasia and gastric carcinoma. The distribution of PHA and PNA receptors was correlated with the histologic type and the degree of differentiation. 81.3% lymph node metastatic tumors produced cellular glycoconjugates, which differed in lectin binding profiles from those made by the cells in the respectively primary carcinoma. More PHA receptors were produced by the lymph node metastatic tumors than primary carcinoma. PMID- 8221248 TI - [The relation between the volume of gastric mucosal blood flow and acid secretion in patients with gastric and duodenal ulcer]. AB - The relationship between gastric mucosal blood flow and acid secretion was studied in 39 patients. In the active stage of gastric ulcer, the volume of gastric mucosal blood flow (GMBF) was decreased, and that of GMBF at the ulcer margin was decreased significantly and increased markedly at the healing stage. The volume of gastric acid output showed no significant difference between the patients with gastric ulcer and controls. The volume of GMBF was slightly increased at all stages of duodenal ulcer. The volume of gastric acid output was increased in the patients with duodenal ulcer. A positive relationship was noted between the volume of maximal acid output and GMBF in the patients with duodenal ulcer. The results indicated that there is a great difference between gastric ulcer and duodenal ulcer in the functions of mucosal microcirculation and acid secretion cells. The reduction of mucosal protective function may be attributable to the pathogenesis of gastric ulcer. The strengthening of the injury factors plays an important role in causing duodenal ulcer. Increased mucosal blood supply may promote the healing of the ulcer. PMID- 8221249 TI - [Effects of anisodamine and ampicillin on cerebrospinal fluid in experimental E coli meningitis]. AB - 24 hr and 27 hr after K1-positive E coli suspended in 0.25 ml of saline (E coli 8 x 10(7)/ml) was injected into the cisterna magna of Chinese rabbits, the levels of bacterial titer, endotoxin, malondialdehyde (MDA), lactate and lactic dehydrogenase (LDH) of the cerebrospinal fluid (CSF) were increased significantly. After ampicillin therapy alone for 3 hr, the level of CSF bacterial titer was decreased and that of LDH increased, but the others didn't change significantly. After anisodamine and ampicillin therapy for 3 hr, the levels of CSF endotoxin and lactate concentrations were decreased from 3.2 +/- 0.6 to 2.1 +/- 0.9 log 10 IU/ml and from 8.6 +/- 2.0 to 6.5 +/- 1.8 mmol/L respectively. The CSF MDA concentration (0.31 +/- 0.07) was lower than that of the nontreatment group (0.59 +/- 0.25) or the ampicillin treatment group (0.59 +/ 0.21 nmol/ml) (P < 0.01 or 0.05). These results suggest that anisodamine combined with antibacterial therapy for experimental E coli meningitis is superior to antibacterial treatment in efficacy. PMID- 8221250 TI - [Hyperselective posterior rhizotomy in the treatment of spasticity of paralytic limbs]. AB - 108 cases of spasticity of paralytic limbs were treated successfully with hyperselective posterior rhizotomy (SPR). Among them, 100 had cerebral palsy, 2 hemiplegia, 2 paraplegia, 3 sequelae of cerebral injury, and 1 multiple sclerosis. Cervical SPR was performed in 12 cases and lumbosacral SPR in 96 cases. After laminectomy, the posterior nerve roots were split into some rootlets, and the lower threshold rootlets were divided after electrical stimulation. Follow up for 6-30 months showed an effective rate of 95% and an improvement rate of 81%. PMID- 8221251 TI - [X-ray computed tomography in the diagnosis of peritoneal and retroperitoneal neoplasms]. AB - CT findings of peritoneal and retroperitoneal neoplasms were evaluated in 24 patients. The accuracy of CT in locating peritoneal and retroperitoneal tumors was 87% and in differentiating benign from malignant ones 96%. It is difficult to evaluate the tumors histopathologically. We suggest that CT scan be preferable in diagnosis of tumors in that area. PMID- 8221252 TI - [Correct use of the techniques of tritium labeled deoxythymidine incorporation into deoxyribonucleic acid]. PMID- 8221253 TI - Pollens and allergic disease: do not overlook your own backyard. PMID- 8221254 TI - Hypereosinophilic syndromes: association with vasculitis, fibrosis and autoimmunity. PMID- 8221255 TI - Regulation and functions of stress proteins in allergy and inflammation. PMID- 8221256 TI - Liposomes in allergy and immunology. PMID- 8221257 TI - Total blood eosinophils, serum eosinophil cationic protein and eosinophil protein X in childhood asthma: relation to disease status and therapy. AB - Blood eosinophils, and serum levels of the eosinophil proteins, eosinophil cationic protein (ECP) and eosinophil protein X (EPX) were measured in childhood asthma. Seventeen patients mean age 11.9 years who were symptomatic with asthma, were enrolled in a study examining the eosinophil counts and eosinophil proteins at the onset of study and after treatment in relation to changes in their baseline forced expiratory volume at 1 second (FEV1) and % predicted FEV1. The patients with symptomatic asthma were compared with 17 patients mean age 12.0 years with asymptomatic asthma maintained on daily inhaled steroid and 13 patients, mean age 12.0 years, without asthma but with urticaria who served as non-asthma controls. Patients with symptomatic asthma did not have significantly higher initial eosinophil counts compared with those with asymptomatic asthma (0.43 x 10(9)/l vs 0.26 x 10(9)/l, P = 0.09) but had higher serum ECP levels (28.9 micrograms/l vs 18.5 micrograms/l). Both asthma patient groups had significantly higher serum ECP levels (P < 0.01) than the controls (9.8 micrograms/l). After therapy consisting of increased dose of inhaled steroids and/or oral steroids, patients in the symptomatic asthma group demonstrated a significant rise in FEV1 (1.67 l/sec at Visit 1 vs 2.08 l/sec at Visit 2, P < 0.001). A similar rise was seen for % predicted FEV1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221258 TI - Nodules, eosinophilia, rheumatism, dermatitis and swelling (NERDS): a novel eosinophilic disorder. AB - This study presents the clinical and laboratory findings of a novel syndrome associated with eosinophilia. Two young women presented with marked eosinophilia, and large, non-tender compressible articular nodules arising from the tenosynovium of extensor tendons, dermatitis, episodic swelling of the hands and/or feet and pain in adjacent muscles and joints. Tissue specimens were examined by routine haematoxylin and eosin staining, immunofluorescent staining for eosinophil granule major basic protein (MBP) and rhodamine-avidin or tryptase staining for mast cells. Plasma levels of MBP and eosinophil-derived neurotoxin (EDN) were quantitated by immunoassay. The first patient presented in 1967 at the age of 20 and had, in addition to nodules and eosinophilia, dermographism, recurrent episcleritis and axillary urticaria. Biopsy of a nodule showed tenosynovitis with necrotizing granulomas, non-specific vasculitis, eosinophils and eosinophil degranulation as shown by extracellular deposition of eosinophil granule MBP. Her symptoms responded to low-dose, alternate-day prednisone and have remained quiescent over the past 15 yr. The second patient presented in 1990 at the age of 28 with generalized pruritic dermatitis for 15 yr, eosinophilia for 2 yr, subcutaneous nodules and non-limiting pain in several joints. Biopsy of a nodule showed chronic mild tenosynovitis, numerous eosinophils and extracellular deposition of MBP. She remains untreated. Serum IgE values and plasma levels of MBP and EDN were elevated in both patients; mast cells were numerous in their synovial tissue. Based on their clinical courses, these patients reveal the existence of a distinctive, relatively benign eosinophilic disorder with good long-term prognosis. PMID- 8221259 TI - Gastrointestinal stability of baker's yeast allergens: an in vitro study. AB - An in vitro model was established to study the stability of baker's yeast (Saccharomyces cerevisiae) allergens in conditions simulating the gastrointestinal tract. The protocol consisted of 2 hr incubation under gastric conditions (pH 1.2, +37 degrees C and gastric enzymes) and 2 hr incubation under duodenal conditions (pH 6.8, +37 degrees C and duodenal enzymes). These were studied together and separately, as well as under pure acidic conditions without gastric enzymes. The yeast extracts contained equal amounts of allergen and were analyzed by IgE-immunoblotting. The acidic conditions had partly an enhancing and slightly degrading effect on the yeast allergens, whereas the gastric enzymes destroyed several allergens, including the important intermediate allergens of 31 and 45 kD. After treatment under both gastric and duodenal conditions most of the yeast allergens were destroyed, except mannan and a 10 kD protein component. The findings suggest that the allergen exposure caused by baker's yeast takes place mainly on the mucosal surfaces orally and oesophageally and through viable baker's yeast organisms that manage to pass the stomach and duodenum and possibly lead to intestinal growth of the organism. Patients with IgE production against the 10 kD allergen and mannan are, however, moderately exposed to allergens consisting of soluble antigenic material only. PMID- 8221261 TI - Mite allergen content in mattress dust of Dermatophagoides-allergic asthmatics/rhinitics and matched controls. AB - It has been suggested that the mites Dermatophagoides pteronyssinus and D. farinae are important indoor environmental factors facilitating both the sensitization of atopic subjects and asthmatic attacks of house dust-sensitive patients. Contradictory results have been reported about the current exposure to mites or their allergens among patients and control groups of atopic or non atopic subjects. In order to determine whether there is a difference in mite exposure levels between D. pteronyssinus-sensitized asthmatics and/or rhinitis and control subjects we considered a case-control study of 70 patients with asthma and/or rhinitis and positive skin test reactions to D. pteronyssinus and twice as many control subjects who were matched as to age and sex. The first control subject for each patient was an immediate neighbour of the patient and the second was patient arbitrarily chosen among hospitalized patients. Mite allergen exposure was measured in mattress dust collected under standardized conditions, by measuring Der p I+Der fI content and by performing a semiquantitative guanine determination (Acarex-test). The content of Der p I and Der fI was very high both in the homes of patients and those of healthy individuals: 69% of the total samples contained more than 10 micrograms Der pI+Der fI/g of dust. There was no significant difference between cases and controls with respect to Der p I, Der fI, Der p I+Der fI content and Acare class distributions. The calculated odds-ratios associated with the Acarex test and the mite allergens did not differ significantly from the level 1.0.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221260 TI - Sensitization to four common inhalant allergens within 302 nuclear families. AB - The coincidence of allergic sensitization was investigated in 302 school-aged children and their parents. Specific sensitization to four common inhalant allergens (grass and birch pollens, cat dander, Dermatophagoides pteronyssinus) was ascertained by means of skin-prick tests (SPT) carried out on the complete family unit at the beginning of a 22-month follow-up period. The same test procedure was then repeated on the children twice at 11-month intervals to provide cumulative prevalences of sensitization. A clinical history of atopy in the children (hay fever or asthma; n = 47), which was derived from an interview, is associated with sensitization (positive SPT in 89%). For three allergens (grass and birch pollens, cat dander) sensitization occurs significantly more frequently in the children of mothers who are sensitized to the same allergen (odds-ratios (ORs), 2.5-4.1). Additionally, in three of the four explanatory models related to a single antigen, maternal sensitization to one of the complementary allergens is of importance (ORs, 2.7-3.7). In contrast to this finding, none of the paternal sensitizations has statistical significance. Based on a reaction to at least one of the four allergens, the child's relative risk to be sensitized is increased in case of maternal (OR, 2.88; P = 0.001) but not of paternal (OR, 1.06; P = 0.83) sensitization. In conclusion, our data indicate that the maternal status is more predictive than that of the father with regard to the child's risk of sensitization. PMID- 8221262 TI - CD23 expression on B-lymphocytes and its modulation by cytokines in allergic patients. AB - The aim of this study was to assess the expression of CD23 on peripheral blood B cells, and its in vitro modulation by recombinant human interferon-gamma (IFN gamma) in phytohaemagglutinin-(PHA) or recombinant human interleukin-4 (IL-4) stimulated cultures in atopic patients with Dermatophagoides pteronyssinus hypersensitivity and in healthy non-atopic subjects. Atopic patients with asthma not receiving allergen-specific immunotherapy (n = 21) were studied and further compared with a group of atopic subjects with asthma under allergen-specific immunotherapy (n = 21). They were age-(+/- 5 yr) and sex-matched. The results were also compared with those obtained in the non-atopic group (n = 11). CD23 expression on B-lymphocytes and its modulation were analyzed by flow cytometry using conjugated monoclonal antibodies with a double immunofluorescence method. Atopic patients had an increase in the percentage of B-cells expressing CD23 in peripheral blood. Phytohaemagglutinin and IL-4 induced a rise in the percentage of CD23-positive B-cells in both atopic groups and non-atopic subjects. Phytohaemagglutinin provoked an increase in the intensity of CD23 expression on B cells from stimulated cultures in all groups, while IL-4 only produced a significant increase in atopic patients. The presence of IFN-gamma decreased the CD23 expression on B-cells in PHA-stimulated culture of atopic patients, whereas it caused an increase in CD23 expression in the non-atopic group. Furthermore, the presence of IFN-gamma in IL-4-stimulated cultures induced a decrease in CD23 expression on B-cells in all cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221264 TI - Cough, wheeze and asthma diagnosis in children. PMID- 8221263 TI - Toxicity and immediate allergenicity of latex gloves. AB - Surgical latex gloves and a natural rubber latex (NRL) catheter were evaluated for biocompatibility by cell culture methods and Type I allergic reactions in operating room personnel using latex gloves. The prevalence of relevant immediate allergy to NRL was 5.2% when using eluates made from different latex gloves as allergens and the skin-prick test methodology. The reliability of a questionnaire in detecting the sensitized persons was limited. Weak positive skin test reactions to surgical latex glove eluates were found to be unreliable as compared with challenging with the corresponding glove. Latex gloves exhibited more toxicity in cell culture than non latex gloves, but, inversely, the toxic catheter gave weak positive skin-prick test reaction in only one sensitized person, confirming that the allergenicity and toxicity of natural rubber products do not refer to the same properties of the products. Catheters exhibiting toxicity in cell culture have earlier been found to cause significant clinical symptoms in operated patients, but in the case of surgical gloves this relationship is not clear. One glove brand with a low content of known sensitizing rubber chemicals caused clear toxicity in cell culture but was well tolerated by the hospital personnel. PMID- 8221265 TI - The risk of allergy related to general anaesthesia. PMID- 8221266 TI - Priorities for understanding and managing occupational allergy: a Delphi consensus. AB - Limiting factors in the understanding, clinical management, and prevention of occupationally related allergic disease should be identified, since these dictate the tasks that need to be undertaken and/or important research questions that warrant an answer. Through an adaptation of the Delphi technique, a working group of the Special Interest Group on Occupational Allergy of the British Society of Allergy and Clinical Immunology addressed these issues. The consensus categorized and ranked the limiting factors and tasks needed in relation to epidemiology and natural history, mechanisms, clinical diagnosis, statute and education, and the workplace environment. This report should help focus the debate regarding which issues are perceived to be most relevant and timely for research and good practice in the field of occupational allergy. PMID- 8221267 TI - Standardization of pollen allergens of Parthenium hysterophorus and selection of an in-house reference extract. AB - A standardized in-house reference extract from the pollen of Parthenium hysterophorus, which is responsible for the high incidence of allergic rhinitis in India, was generated and examined by skin test, radio-allergosorbent test inhibition and isoelectric focusing. Parthenium reference allergen discs and positive reference serum were also generated. These reference reagents could not only be used for the quantitation of Parthenium-specific IgE in the sera of rhinitis patients but also for the evaluation of allergenic activity (relative potency and lot-to-lot variation) of different batches of Parthenium pollen. PMID- 8221268 TI - Venom-specific IgG antibodies in bee and wasp allergy: lack of correlation with protection from stings. AB - This paper investigates the relationship between venom IgG levels and protection from stings. Venom-specific IgG antibody levels have been measured by radioimmunoassay in untreated wasp-(n = 38) and bee-allergic (n = 16) patients presenting with systemic reactions to stings and in a sub-group of these (wasp = 15; bee = 9), before and after the initial course of venom immunotherapy (VIT). A history was taken of all reactions, the last systemic reaction being graded on a scale of 1-8 and of the number and timing of stings. In untreated patients venom IgG levels were much higher in bee-allergic patients (mean +/- s.e. = 68.2 +/- 7.1% positive pool) than in the wasp group (27.1 +/- 4.2%) (P < 0.05 Mann-Whitney U-test). There was a marked rise in venom IgG after the initial course of VIT in the wasp group (geometric mean and 95% confidence intervals = 40.5%, 28.8-54.3) but a much smaller rise in the bee group (15.3%, 6.6-24.1), with no overlap in the 95% confidence intervals. Bee patients, who were mainly beekeepers or their relatives, had been more heavily immunized with venom than wasp patients. They had received: (i) more stings (mean number of stings: bee, 26; wasp, 4; P < 0.001) and (ii) more stings per year. Wasp patients received their smaller number of stings over a much longer period, up to 40 yr. There was no correlation between the severity of the last systemic reaction and the venom IgG levels alone or venom IgG and IgE levels in combined analysis in either bee or wasp patients. This study shows that the pattern of IgG response differs in bee and wasp allergic subjects, and that most bee-allergic subjects with systemic reactions have high levels of venom IgG. The degree of immunization with venom seems to be an important determinant of the venom IgG level. Our findings suggest that venom specific IgG levels do not predict systemic reactions to stings and are not useful for monitoring VIT. If protection from stings is IgG-mediated, our observations suggest that the relevant immune response is more complex, possibly involving IgG sub-classes, IgG antibodies to individual venom antigens or antibody affinity, and not adequately reflected by measurement of the concentration of venom-specific IgG. PMID- 8221269 TI - Lymphocyte transformation test with house dust mite (Dermatophagoides pteronyssinus) in normal children, asthmatic children and asthmatic children receiving hyposensitization. AB - In the first part of this study the proliferative response of lymphocytes (lymphocyte transformation test) to house dust mite (HDM) stimulation in cultures was studied in normal children (n = 16), asthmatic children who never received hyposensitization (HS) (n = 50) and asthmatic children receiving HS with HDM for at least 6 months (n = 20). The results are expressed as disintegrations per minute (d.p.m.) and as stimulation index (SI = d.p.m. in the presence of the allergen/d.p.m. in the control culture). A positive SI (> 2) was found in 54% of the asthmatic children who never received HS, in 30% of the asthmatics receiving HS and in none of the normal children. Furthermore, between asthmatics with and without HS, the SI was not statistically different, although asthmatics without HS tended to have a higher SI (median value: 2.13 vs 1.38) (P = 0.10). In a second series of experiments the effect of adding interleukin-2 (IL-2) to the lymphocyte cell culture was studied in asthmatic children with and without HS. Interleukin-2 induced an additional stimulatory effect on the lymphoproliferative response to HDM and to phytohaemagglutinin in patients who never received HS, but had no effect in patients receiving HS. We conclude that HS treatment seems to have an inhibiting effect upon this proliferative response, not only inhibiting the degree of the allergen-induced lymphocyte proliferation, but also inhibiting the sensitivity of proliferating lymphocytes for IL-2. These inhibiting effects upon lymphocytic activation could be responsible for the anti-inflammatory effects (i.e. suppression of the late asthmatic reaction) of HS. PMID- 8221270 TI - Increased immune reactivity to house dust mites in adults with chronic rhinosinusitis. AB - Sixty-three adults with symptomatic chronic rhinosinusitis had computerized tomographic (CT) scans of the paranasal sinuses, which were used to quantify disease severity. These patients were divided into three closely age- and sex matched groups: a CT scan-negative group (chronic rhinitis only), a mild sinusitis group and a severe sinusitis group. Serum dust mite-specific IgG levels were found to be significantly elevated in the sinusitis patients compared with a matched group of asymptomatic normal individuals. Levels were highest in the more severe sinusitis group, in which the mean titre was 559 U/ml and the incidence of titres greater than 400 U/ml was 48%, as compared with a mean titre of 139 U/ml and only a 10% incidence of titres greater than 400 U/ml in the normal subjects (P < 0.005 and < 0.01). In contrast, although the frequency of immediate hypersensitivity to dust mite, as assessed by intradermal skin tests, tended to be higher in patients with sinusitis, it was not significantly different from normal individuals. The association between mite IgG and disease was most striking in the patient sub-group with negative mite skin tests. In this group, mite IgG levels were significantly higher than normal, even in those patients with only chronic rhinitis. These findings demonstrate that increased serum levels of IgG against dust mites are strongly associated with chronic rhinosinusitis, especially in the sub-group of patients who are not allergic to mites. PMID- 8221271 TI - The determinants of airway hyperresponsiveness to hypertonic saline in atopic asthma in vivo. Relationship with sub-populations of peripheral blood leucocytes. AB - In patients with asthma there is only a weak relationship between airway responsiveness to hypertonic saline and methacholine. We tested the hypothesis that airway responsiveness to hypertonic saline in asthma is related to the presence and activity of inflammatory cells in the peripheral blood. Nineteen atopic asthmatic adults (19-28 yr; PC20 0.06-12.4 mg/ml), not receiving steroid treatment, entered a metacholine and hypertonic saline period in random order. Dose-response curves to doubling doses of inhaled methacholine (0.03-256 mg/ml) or hypertonic saline (0.9-14.4% NaCl) were obtained twice in each period, 7 days apart. The response was measured by FEV1. Methacholine responsiveness was measured by PC20 METH of FEV1 and responsiveness to hypertonic saline was expressed as the percentage fall in FEV1 after 14.4% NaCl (HYP14.4%). Peripheral blood was collected before the second challenge test of each period. Apart from leucocyte counts and serum eosinophilic cationic protein (ECP) level, sub-sets of lymphocytes (CD4+/CD3+, CD8+/CD3+, CD25+/CD4+ and VLA-1+/CD4+) were determined using flow cytometry. HYP14.4% was positively correlated to basophil, eosinophil and monocyte counts (r = 0.64, 0.54 and 0.44, respectively; P < 0.05). The basophil count remained positively related to HYP14.4% when PC20METH or FEV1%pred were entered in multiple linear regression analyses (r = 0.66 and 0.75, respectively; P < 0.05). There were no significant relationships between HYP14.4% or PC20METH on one side and ECP level or T-lymphocyte subsets on the other (P > 0.05). We conclude that airway responsiveness to hypertonic saline is positively related to the number of peripheral blood basophils, eosinophils and monocytes. Basophil count is an independent correlate of responsiveness to hypertonic saline, after correction for methacholine responsiveness and baseline lung function. This fits in with active involvement of basophils in airway narrowing to hypertonic saline in vivo. PMID- 8221272 TI - Cough variant asthma is associated with a higher wheezing threshold than classic asthma. AB - Cough variant asthma is an occult form of asthma in which the only sign or symptom is chronic cough. It is a common problem amongst all ages that frequently goes unrecognized, leading to underdiagnosis and undertreatment. To characterize the pathophysiological abnormalities in these patients, we performed bronchial provocation tests with methacholine using the stepwise doubling concentration technique, and measured the concentration of methacholine and the severity of airway obstruction when wheezing was first detected. Airway hyperresponsiveness, defined as PC20 in the cough variant asthma group was not significantly different from that of classic asthma. There was a good correlation between the PCW (the concentration of methacholine causing wheezing) and the PC20 with the PCW values higher than PC20 values in both groups. However, in the cough variant asthma group, the PCW:PC20 ratio was greater than that of the classic asthma group. Furthermore, the mean % fall in FEV1 at which wheezing was first detected in the former group was significantly larger than that of the latter group. The results indicated that the mechanism for the manifestation of cough without wheeze in the cough variant asthma may be a higher wheezing threshold, i.e. wheezing becomes audible at the greater degree of airway obstruction than classic asthma. They suggested that patients with cough variant asthma may represent a subset of asthmatic subjects whose airways are less able to produce a wheeze. PMID- 8221274 TI - Encopresis: developmental, behavioral and physiological considerations for treatment (continuing education credit). PMID- 8221273 TI - Allergen extracts. Standardization of preparations for bronchial provocation tests. A position paper. (EAACI Sub-committee on Bronchial Provocation Tests). AB - Standardization of allergen provocation tests of the airways requires standardization of each of the steps involved, including standardization of the extracts used. The value of international standards is emphasized and so is the determination of a clinically relevant potency. The allergen extracts used for bronchial provocation tests must be compared with qualitatively and quantitatively clinically relevant standards using complementary immunochemical and biological methods. Ideally the same extracts should be used for diagnosis (skin test, specific IgE determinations, provocations) and treatment (immunotherapy). The importance of the biological compared with the non biological methods is that the biological methods are established in allergenic patients in order to select a clinically relevant potency of the allergen in question, and this potency ought to be confirmed in clinical trials. Once established the biological potency of an in-house preparation can be reproduced by supplementary in vitro methods. Very few commercial allergen preparations are available which fulfil the above-mentioned criteria. PMID- 8221275 TI - Clinton plan: CNA praises access goal, seeks care standards, disclosure laws. PMID- 8221276 TI - Hospital restructuring: the impacT on RNs and patient care. PMID- 8221277 TI - A guarantee--is it ethical? PMID- 8221278 TI - CDIP malpractice insurance is cost effective and competitive. PMID- 8221279 TI - The whole truth or nothing?, a tale of misinformed consent. PMID- 8221280 TI - Achieving success: moving beyond the comfort level. PMID- 8221281 TI - Access and isolation problem solving in endodontics: posterior teeth. AB - Through the pictorial presentation of case studies, this paper explores the versatility of alternative methods of retention as a means of improving overall access to the endodontically-involved tooth. Successful endodontics is the result of precise biomechanical instrumentation and involves procedural manipulations that depend on a complete and unobstructed approach to the working field. This paper is aimed principally at those practitioners who would ordinarily reject rubber dam isolation in cases where the clinical pretreatment conditions are anything but ideal. PMID- 8221282 TI - Down's syndrome and Alzheimer's disease. AB - Individuals with Down's syndrome (DS) who live to be 40 years of age will demonstrate neuropathological changes that are consistent with Alzheimer's disease (AD). Due to modern medical intervention, we are now observing an aging DS population. Middle-aged Down's syndrome adults are actually considered to be "very old," and it is not uncommon to observe a progressive loss of cognitive function and a decline in the ability to perform daily tasks consistent with that seen in Alzheimer's disease. At this stage, the DS individual will not be able to perform daily preventive dental care and may be unable to cooperate for professional dental care. Clinicians who care for DS adults must be aware of this problem when preparing their dental treatment plans, which must emphasize preventive care prior to the onset of dementia and the maintenance of that program during their patients' cognitive decline. In the latter stages of AD, it may be necessary to extract all the remaining teeth due to the inability of the individual or care giver to provide adequate oral hygiene to prevent dental caries or periodontal disease. PMID- 8221283 TI - Oral health and the quality of life among older adults: the oral health impact profile. AB - A number of studies are beginning to show that oral disorders can have a significant impact on the functional, social and psychological well-being of older adults. The Oral Health Impact Profile (OHIP) was developed to collect information on the nature and extent of this impact, and to facilitate the use of such data in oral health surveys, the evaluation of dental procedures and the clinical evaluation of patients. This paper describes OHIP and the results of two studies undertaken in Ontario to assess its measurement properties and to provide preliminary data on the way in which oral conditions compromise the quality of life of older adults. The measure proved to be reliable and valid, while data collected using this measure indicated that oral conditions have a negative impact on the daily lives of substantial proportions of older people. This impact was particularly marked among both edentulous and dentulous individuals who did not make regular visits for dental care. PMID- 8221284 TI - [A case of swallowing a lower partial denture]. AB - This paper reports on the case of an elderly person who accidentally swallowed his partial denture. The ingestion did not result in any signs or symptoms and the denture was found totally by chance. A major surgical procedure was required to remove the prosthesis. PMID- 8221285 TI - Successful mandibular anesthesia following numerous unsuccessful attempts: a case report. AB - The management of a very anxious patient who had undergone uneventful dental treatment in the past, but posed a problem when clinicians attempted to secure profound mandibular anesthesia, is discussed. This case report also examines the advantages and disadvantages of the Gow-Gates mandibular block technique, and describes the patient's subsequent management. PMID- 8221286 TI - Do we need "socialized dentistry" in the nineties? PMID- 8221287 TI - Endothelium-derived contracting factors. AB - In the past decade, the importance of endothelium in modulating vascular tone has become apparent. Endothelium can produce different vasodilators, such as prostacyclin, endothelium-derived relaxing factor (most likely nitric oxide), and endothelium-derived hyperpolarizing factor. Endothelium can also produce endothelium-derived contracting factors (EDCF) which can induce constriction of the underlying vascular smooth muscle. Constriction may be mediated by a variety of products released by the endothelium. Thus, by responding to various stimuli and releasing either relaxing or contracting factors, the endothelium can set and modulate vascular tone. PMID- 8221288 TI - The antimicrobial activity of imipenem/cilastatin and its treatment in critical ill patients with polymicrobial and mixed infection. AB - The antimicrobial activity of imipenem/cilastatin (IPM/CS) was determined for a broad spectrum of 7,157 organisms isolated from the Clinical Microbiology Laboratory of Chang Gung Memorial Hospital, Linkou Medical Center, between April 1 and June 30, 1988. Ninety eight point one percent of 4,389 gram negative aerobes, 95.8% of 2391 gram positive rods, and 95.9% of 507 anaerobes, were shown to be sensitive to IPM/CS. Ninety nine point two percent of 837 Pseudomonas aeruginosa isolates were sensitive to this antibiotic. This study also disclosed that this agent was much more active against Pseudomonas aeruginosa than any other tested aminoglycosides (gentamicin, amikacin, netilmicin) or third generation cephalosporins (cefotaxime, latamoxef, ceftazidime). Twelve critically ill patients with polymicrobial and mixed infection were recruited into this trial. All patients received IPM/CS 500 mg intravenously every six hours except one patient with poor renal function had 250 mg every six hours. The average duration of therapy was 12.5 days. All patients were evaluated according to the selection criteria. IPM/CS achieved favorable clinical response in 83.7 percent. The antibiotic was well tolerated. One patient discontinued treatment because of jaundice. One patient had superinfection of fungemia. The results suggested that IPM/CS is very useful in the treatment of patients with mixed infections due to gram positive, gram negative, and anaerobic bacteriae, even when empirical therapy with other antibiotics fails. PMID- 8221289 TI - Electrodiagnostic studies in myasthenia gravis. AB - Forty-three patients with myasthenia gravis (MG) were studied to compare the sensitivity of repetitive nerve stimulation (RNS) test with single fiber electromyography (SFEMG) in the diagnosis of MG. SFEMG was abnormal in 81% (35 of 43 cases), while RNS test was abnormal in 53% (23 of 43 cases). In ocular MG, SFEMG showed a much higher positive rate (70%, 16 of 23 cases) than RNS test (22%, 5 of 23 cases). Both studies had similar abnormal rates in the generalized group. In patients studied with SFEMG in the EDC muscle, the clinical severity was closely correlated with jitter abnormality. We conclude that SFEMG is more sensitive than RNS test in the diagnosis of MG, especially in the ocular group. PMID- 8221290 TI - Features of spinal cord injury in Taiwan (1977-1989). AB - In order to establish an etiological and statistical base for spinal cord injuries, 1,617 spinal cord injured patients admitted to the Chang Gung Memorial Hospital in Taiwan during the period of 1977 to 1989 were reviewed. The most common causes of injury were pedestrian (29.31%) and motorcycle (28.88%) accidents. The greatest incidence of injury was in the 26-35 year age group. The complete tetraplegic patients had the highest mortality rate (26.5%). Additional features studied were the time of occurrence and pattern of injury. Information gathered from this study suggest the need to establish a Spinal Cord Injury Prevention Program, to develop a Prehospital Care System and set up comprehensive Spinal Cord Injury Units in Taiwan. We expect this study to be adaptable to other similar developing countries. PMID- 8221291 TI - Planned replacement of daily wear Acuvue disposable contact lenses. AB - A study of Acuvue disposable contact lenses was performed on 30 patients. The lenses were worn daily and renewed every 2 weeks for 6 months. During collection of these patients, another 16 were excluded due to an improper lens fit. Of the 30 patients included in the study, 98 to 100% had good vision and felt comfortable when wearing Acuvue lens during the whole study course. Twelve (40%) patients felt the lens were floppy and had some difficulty handling them at first. Nineteen (2.4%) of the 780 lenses used had prominent defect and 4 (0.5%) were lost during daily manipulation or wearing. Four (6.6%) eyes suffered mild punctate superficial keratitis. So, patients should be selected carefully because Acuvue lens only have one size and base curve. Acuvue lens can provide good vision and comfort of lens wearing but handling may be difficult at first. Caution should be exercised in checking and manipulating lens to prevent lens and ocular damage. PMID- 8221292 TI - [Acute upper gastrointestinal bleeding in Chang Gung Memorial Hospital: comparison between 1980 and 1989]. AB - Upper gastrointestinal (UGI) bleeding remains a great medical problem despite the improvement in both diagnostic and therapeutic management. We retrospectively analyzed 560 cases (male 429 cases, female 131 cases, mean age 45.8 +/- 23.1 years) of acute UGI bleeding within one year from January to December 1980, and 1872 cases (male 1395 cases, female 477 cases, mean age 48.7 +/- 27.5 years) within one year from January to December 1989, in order to define the changing pattern of etiology in the past 10 years. The major cause of bleeding was duodenal ulcer bleeding, which occurred at the age of 20-40 years. The incidence significantly decreased (57.8% in 1980 VS. 46.3% in 1989, P < 0.001). The second common cause was gastric ulcer (11.1% VS. 13.5%), which occurred mainly at the age of 50-70 years. The third was esophageal varices bleeding with a significantly increased incidence (6.6% VS. 11.4%, P < 0.001), which occurred mainly at the age of 40-60 years. The other less common causes included gastric cancer (5.9% VS. 5.8%), which occurred mainly at the age of 50-70 years, gastric erosion (5.2% VS. 6.1%), Mallory Weiss tear (2.1% VS. 3.1%), esophagitis (1.9% VS. 2.9%), Dieulafoy's ulcer (1.6% VS. 2.7%), vascular lesion (1.6% VS. 2.6%), and non-diagnostic cases (6.2% VS. 5.6%). The ratio of male to female for each etiology of UGI bleeding was about 3 to 1 in both 1980 and 1989.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221293 TI - [Pheochromocytoma nuclear pattern analysis by flow cytometry]. AB - Flow cytometric nuclear DNA analyses were performed on paraffin-embedded tissue samples taken from 39 patients with pheochromocytoma treated between 1981 and 1992. The Hedley technique was used for measurement of nuclear DNA contents. Only 30 cases out of the 39 tumors were available for the determination of DNA ploidy analyses. Ten cases (33.3%) showed a diploidy pattern, 8-cases (27%) showed an aneuploidy pattern and 12 cases (40%) revealed a tetraploidy pattern. Four cases of malignant pheochromocytoma with regional or distal metastasis had been noted during a mean follow-up period of 4.75 years. All these four cases showed tetraploidy nuclear pattern with vascular invasion on pathological examination, which was statistically significant (p < 0.001) in differentiation between benign and malignant pheochromocytoma. About one third of patients with tetraploidy appeared to have malignant tendencies and this was statistically significant (p < 0.05). None of the patients with either diploidy or aneuploidy pattern had disease progression. The location of the tumor (adrenal vs. extra-adrenal) and 24 hours urine total vanillylmandelic acid (VMA) amounts were not significantly different from the diploid, aneuploid or tetraploid groups. These results suggested that nuclear DNA ploidy pattern is an important independent, and useful prognostic factor for patients with pheochromocytoma. PMID- 8221294 TI - Type I familial amyloid polyneuropathy--report of a family in Taiwan. AB - We report three cases of histologically verified systemic amyloidosis with polyneuropathy. Common to them were early onset progressive peripheral sensorimotor disturbance starting in the legs and prominent autonomic dysfunctions such as postural hypotension, anhidrosis, and loss of pupillary light reflexes. Other characteristic features included vomiting, alternating diarrhea and constipation, opacities of the vitreous bodies, and congestive heart failure. All these clinical manifestations resemble type I familial amyloid polyneuropathy described by Andrade in 1952 from Portugal. Sural nerve biopsy stained with Congo red showed typical green birefringence under polarized light microscope. Histologically verified familial cases of this form of amyloid polyneuropathy have not been reported from Taiwan before. PMID- 8221295 TI - Deep vein thrombosis during pregnancy. AB - Antepartum deep vein thrombosis is quite rare in Chinese women during pregnancy. An adequate diagnosis and early treatment are extremely important as the deep vein thrombosis can embolize, in particular, to the pulmonary vasculature. We present a case of pregnancy at 36 weeks gestation complicated by deep vein thrombosis. The objective diagnosis of deep vein thrombosis for obstetric patients is the noninvasive Doppler ultrasound and plethysmography. An early diagnosis is important to reduce maternal and fetal risks related to embolization. Continuous therapy of low dose intravenous heparin is safe and effective during pregnancy. PMID- 8221296 TI - [Squamous cell carcinoma originating from ovarian dermoid cyst--report of three cases]. AB - We report three cases of squamous cell carcinoma originating from ovarian dermoid cysts. All patients were postmenopausal with the chief complaints of lower abdominal pain and palpable masses found by themselves. Two of them had diabetes mellitus. The tumor sizes were all more than 10 cm. The diagnoses were made by histological examination of tissues removed at surgery. Two cases were categorized into the FIGO stage IIIc and one in stage Ia. Sonographic findings were characterized by a large adnexal mass with mixed components. An elevated serum squamous cell carcinoma antigen was found in the two cases of advanced stage and one of them also had an elevated serum CA-125 level. All reported cases were unilaterally involved without ascites at laparotomy, while omental lymph nodes metastases were noted in the two cases of advanced stage. One of the patients in the advanced stage died six months after surgery and radiation therapy due to recurrence. The other patient in the advanced stage refused further treatment even though recurrence was found five months after surgery and chemotherapy. There was no evidence of recurrence in the early stage case during the two years of follow-up. PMID- 8221297 TI - [A phoniatric treatment report for a dysphonia case with unilateral false cord bulging]. AB - This case report was a 43-year-old male with phonation problem described as hoarseness, reduced loudness and with effort. Patient was examined by a laryngologist and a voice-speech pathologist. The laryngeal finding showed left false cord bulging under indirect laryngoscope and fiberscope. Patient's dysphonia was analyzed from perceptual and acoustic parameters. Patient was diagnosed as dysphonia with left cord bulging and under a phoniatric treatment. The goal of treatment was to eliminate vocal abuse and misuse behaviors, stimulate true cords function by Boone's "inhalation phonation" method then reduce compensatory behaviors of false cord involvement and balance life between work and after work. The results after the phoniatric treatment and underlying phonation mechanism were presented and discussed. PMID- 8221298 TI - [Capability of producing interleukin-2 of lymphocyte subsets and mononuclear leukocytes in burn patients]. PMID- 8221299 TI - [Treatment of chronic osteomyelitis of leg with free latissimus dorsi musculocutaneous flap anastomosed to contralateral leg vessels]. AB - A free latissimus dorsi musculocutaneous flap was first anastomosed to dorsal vessels of the contralateral leg. It was then used to cover a large defect of the leg as a result of chronic osteomyelitis. The use of such a cross-leg flap was not affected by the poor circulatory condition of the diseased leg. It was done in 20 patients with satisfactory results. All of them could take part in the physical work. PMID- 8221300 TI - [The transconjunctival lower blepharoplasty]. PMID- 8221301 TI - [Septicemia in early stage of severe burns]. AB - Among 108 cases of severely burned patients, 11 were complicated with septicemia in early stage. This analysis showed that pathogenic bacteria might mainly come from the intestinal tract, and shock played a critical role in the occurrence of septicemia, and at the same time septicemia aggravated shock and also induced complications of internal organs. Attention should be paid to severely burned patients who were hospitalized late or with severe shock, and early administration of broad-spectrum antibiotic against intestinal flora might improve the therapeutic effect. PMID- 8221302 TI - [Preliminary study of density of Langerhans' cells and distribution of ABH blood group antigen in prepuce]. PMID- 8221303 TI - [Comparative study on tissue morphology and vascular changes of skin xenografts, allografts and autografts after transplantation]. AB - In this study, the tissue morphology and the vascular changes of skin xenografts from the rats to the rabbits were compared with that of skin allografts and autografts simultaneously transplanted to the same recipient by the use of histological examination and dye injection. The results suggested that the xenografts were incapable of becoming vascularized, and exhibited necrosis on the third day, while early vascularization was demonstrated in the allografts and autografts. The xenografts underwent a course of "white graft rejection", which probably contributed to rejection. PMID- 8221305 TI - [An experimental observation of effect of tetrandrine on pulmonary dysfunction in rabbit model of smoke inhalation injury]. AB - A rabbit model with severe smoke inhalation injury was adopted to observe the effect of tetrandrine, a calcium antagonist, on pulmonary dysfunction after smoke inhalation during 24 hours post injury. It was found that tetrandrine could decrease the permeability of pulmonary and tracheal vascular permeability (p < 0.01), reduce lung edema and improve the function of respiration. These results imply that tetrandrine might be used in the treatment of inhalation injury. PMID- 8221304 TI - [Comparison of topical therapeutic effect between silver norfloxacin (Ag-NFL) and silver sulfadiazine (Ag-SD) in treatment of pyocyaneous sepsis in burn rats]. AB - The purpose of the study was to carry out a further investigation of the efficacy of topical antimicrobial treatment of Ag-NFL in comparison with that of the most commonly used topical agent Ag-SD. An experimental model of rats with 20% TBSA full thickness burns seeded with 109 CFU of Pseudomonas aeruginosa ATCC-27853 was used. At 24, 48 and 72h after the seeding bacteriologic examination showed that the subeschar bacterial counts exceeded 10(6) CFU per gram of tissue. Subeschar bacterial counts were zero in the group in which Ag-NFL was used, while those in the group using Ag-SD were almost the same as the control. The amount of silver accumulated in the eschar of Ag-NFL, treated rats was significantly higher than that of Ag-SD treated rats. In conclusion, Ag-NFL was proved valuable in the treatment of burn wound infection caused by invading organisms, particularly by the Ag-SD resistant strain of Pseudomonas. PMID- 8221306 TI - [Research on clinical applied anatomy of breast]. PMID- 8221307 TI - [Contribution to the treatment of the benign deformities of the breast]. PMID- 8221308 TI - [Evolution of plastic surgery reflected in changes in breast reconstruction]. PMID- 8221309 TI - [Liposculpture by syringe technique]. PMID- 8221310 TI - [V-Y advancement of volar skin flap in the repair of fingertip defect and cicatricial contracture]. AB - The traditional Kutler and Atasoy V-Y advancement flaps have minimal range of advancement, so it is not satisfactory to repair large skin defect in fingers, hence limited indications. The sensory function of the repaired fingers is impaired because of injury to sensory nerves and scar formation. Since 1985 to 1991, the V-Y advancement flap with bilateral digital arteries and nerves as the pedicle has been used for the reconstruction of 33 fingertip defects and 5 digital volar cicatricial contracture. Satisfactory results were obtained in all these cases. In this paper, the anatomical characteristics and principles of device were recommended, the advantages and key points to success were discussed. PMID- 8221311 TI - [Early repair of burned cranium]. AB - Early repair was done on 10 patients, of whom there was necrosis of either the outer or both outer and inner tables of the skull as a result of deep burn injuries. Multiple drill holes were made through the necrotic cranial bones until bleeding occurred on 7 PBD. The wounds were immediately covered with skin flaps. A continuous drip of lidocaine, chloramphenicol and normal saline was instituted underneath the flaps. The results after one to five years follow-up were satisfactory. PMID- 8221312 TI - [Correction of naso-labial disfigurement subsequent to bilateral cleft lip repair with combined upper lip flap and Abbe flap]. AB - Anomalies of the nose and lip after repair of bilateral cleft lip and palate is a psychological injury to the patients affecting their social intercourse. The author has designed an operative method using a flap of the fore-lip combined with Abbe flap of the lower lip to repair the anomaly with successful result. In two patients, free Abbe lip flap was used, with one complete success and partial necrosis of the other. PMID- 8221313 TI - [Repair of lower nasal defect with compound skin flap from forehead with trochlear artery and subcutaneous tissue pedicle]. AB - This article reports the repair of lower nasal defect with forehead compound skin flap consisting of the trochlear artery and subcutaneous tissue pedicle in one stage in 11 patients. The repaired area ranged from 2 x 2 to 4 x 3 cm and the frontal bone area 1.5 x 1 cm. This operation had achieved good results, and the frontal bones were not absorbed, except 1 case, after 1 year follow-up. The methods of this flap design and operation were described. The merits, demerits and points for attention were discussed. PMID- 8221314 TI - [Myocardial damage and its mechanism in burn patients]. PMID- 8221315 TI - [Prevention and treatment of common complications following suction lipectomy]. PMID- 8221316 TI - [Suction lipectomy for male gynecomastia]. AB - This paper reports the application of the suction lipectomy to treat gynecomastia in 10 cases with excellent results. A 2-2. 5 cm incision was made on the lower areolar border at 5 to 7 o'clock. The suction power was adjusted to 67-93 kPa. The suction cannula was moved to and fro in radial direction rapidly. The diameter of the sucked area should be larger than desired size of the breast by 2 cm to avoid a saucer deformity. If gynecomastia is entirely due to fat accumulation, suction lipectomy only is enough. But, when gynecomastia is due partly to parenchymal hypertrophy, an additional excision of the parenchymatous tissue is required. After the operation, elastic garment must be worn at all time for 4 to 6 weeks to avoid hematoma and to achieve a flat appearance. PMID- 8221317 TI - [Reconstruction of half of the nose]. PMID- 8221318 TI - [A new scapular free flap with ascending and descending branches of the circumflex scapular artery as the main axial arteries]. AB - Free scapular skin flap was transplanted for repair of burn scars on the face and neck successfully in 8 patients. The ascending and descending branches of the circumflex scapular artery are utilized as the main axial vessel. This method expands the range of use of skin flap in the scapular region. PMID- 8221319 TI - [Oro-pharyngeal flora in severely burned patients]. AB - Throat cultures were taken in thirty-seven cases of burned patients, and also, at the same time, from twenty-three healthy persons. The results showed that the essential oro-pharyngeal flora in healthy persons were Neisseria (65.7%) and Streptococcus viridans (14.3%), and Staphylococcus aureus was obtained occasionally. In twenty-five out of thirty-seven patients dominant growth of invaders was observed. The prominent aerobic bacteria in patients included Serratia (9.33%), Pseudomonas aeruginosa (8.44%) and Proteus (7.11%). The positive rate of fungi was as high as 75.1%, of which Candida was the most commonly found organism. In 7 patients the number of bacteria in the throat was in excess of 108/cm2. Of these 7 patients, 4 died. Quantitative bacterial count of lungs were taken in 3 non-survivors, and the bacterial count was over 10(5)/g. The bacteria in the lungs were as same as the predominantly colonized bacteria in the throat, and in a part of cases were the same as the systemically disseminated bacteria. The study of the throat flora is simple, its result might help the prediction of pulmonary infection. PMID- 8221320 TI - [Comparison of irradiated porcine skin with exposure or occlusive method in the care of second degree burn wounds]. AB - This article compared irradiated porcine skin with exposure and occlusive method in the care of second degree burn wounds. In 30 patients in whom irradiated porcine skin (IPS) was used, no wound infection occurred. The rate of wound infection, in patients on whom herb medicine and exposure method were used, was 19% (7/37), and that of occlusive dressing plus drug group was 22% (2/9). The rate of wound infection of IPS was significantly lower than the other two groups (P < 0.05). The time of wound healing in IPS group was 13.5 + 5.03 days; it was significantly shorter than the other two groups. PMID- 8221321 TI - [Therapeutic effect of calcium preparation in phosphorus poisoning following phosphorus burns]. AB - All of 20 patients with phosphorus burn, admitted from 1987 to 1991, have been cured by intravenous drip of calcium gluconate combined with other therapies. The wounds were washed with tap water, followed by eschar formation in 12 days. The clinical data showed that phosphorus poisoning did not occur if calcium was given very early postburn. Calcium also could abolish or ameliorate clinical manifestations of existing phosphorus poisoning, accelerate the elimination of phosphorus and recovery of damaged organs. PMID- 8221322 TI - [Clinical use of irradiated allogeneic dermis in the coverage of burn wounds]. PMID- 8221323 TI - [An evaluation of problems arising from the use of soft tissue expander]. AB - From November 1987 to July 1992, expanded skin was examined with both light and electron microscopes in 15 patients. Using these results, several problems arising from clinical use of skin expander were evaluated. PMID- 8221324 TI - [Silastic chin implant introduced through an intra-oral small V-shaped incision]. PMID- 8221325 TI - [A clinical research of three dimensional computerized imaging in the aid of operative design for reconstructive surgery of the face]. AB - This article reports the utilization of three dimensional imaging technique of computer in reconstructive surgery of the face. In 312 cases, the technique has been proved that it can make the preoperative analysis computerized, quantitated and picturized. The measurement and analysis through computer are quick and exact, rendering operative designs feasible. The use of the evaluation system can also help enhance the communication and understanding between the surgeon and his patient. The surgical simulation system can replace the traditional preoperative manual work such as paper molds, plaster casts, and wax mold, and it is therefore much valuable for practical use. PMID- 8221326 TI - [Experimental study on the skin by laser-assisted anastomosis]. AB - Incisions in 35 rats were performed on the back bilaterally. Both incisions were sutured by hand with threads at equal interval. One of the incisions immediately welded by laser after suture, the others as follow. When 24 hours after suture, the stitches of the laser welded incisions were removed. The maximal strength force and histology of the bilateral incisions were measured and examined respectively at 1, 3, 7 and 14 days after the operation. The results showed: there was obviously significant differences of the strength force within 3 days between the two incisions. There was slightly inflammatory reactions, no scar and ideal healing in laser welded incisions. PMID- 8221327 TI - [Angiotensin converting enzyme (ACE) in scalded rats and protective effect of chloroquine (CQ) in lung injury]. AB - Wistar rats were scalded and then divided at random into control group and scalded+CQ group. They were sacrificed in lots at 6, 24 and 48 hours after the scald and blood was drawn for the determination of ACE, PLAa, Pa02. Pathological examination was also done. The results revealed that, 1. after scald, the rat's serum ACE was reduced below normal value, and it occurred before changes in blood gas, suggesting that ACE might be used as a valuable supplementary diagnostic criterion for pulmonary failure; 2. CQ provided certain amount of protection in pulmonary failure of scalded rat. PMID- 8221328 TI - [Effect of hydrocortisone, dexamethasone, adenosine triphosphate (ATP), and L glutamine on viability of skin stored at 4-8C]. PMID- 8221329 TI - [Experimental study of pharmacokinetics of cefoperazone (CPZ) in the burn area]. PMID- 8221330 TI - [Effect of burn injury, blast injury and combined burn-blast injury on immune reactions of thymocytes and splenocytes in rats]. AB - Rats were inflicted with burn (15% TBSA full-thickness) by flash thermal radiation, blast injury in shock tube (over pressure 429.3 +/- 11.5 kPa) and both of them, respectively. The dynamic changes of several cellular immune reactions of thymocytes and splenocytes were observed. One hour after injuries, the immune reactions were significantly enhanced in combined injury group as well as in blast injury group, but somewhat depressed in burn group. 12 hours after injuries, the immune reactions in all three groups were obviously depressed. It was found that the degree of depression showed an order as follows: Combined injury > Blast injury > Burn. The combined effects were more serious than that of the sum of two single injuries, and the combined injury recovered more slowly than the other two. Therefore, an aggravated effect was observed in combined injury in this experiment. PMID- 8221331 TI - [Early coverage of deep partial-thickness burn wound with porcine dermal collagen membrane. An experimental study]. AB - Collagen was extracted by pepsin digestion from porcine skin and collagen membrane was prepared by salt precipitation. Collagen membrane as a wound dressing was evaluated in a deep partial-thickness burn wounds in a rat model. Burn wound, 4 x 4 cm, were inflicted by exposing skin to 75 degrees C for 15 seconds followed by de-epithelization. Wound healing was assessed by plenimetry of epithelization on day 10 postwounding. Open wounds exhibited a re epithelialization of 24% of wound area. Collagen membrane dressing significantly improved the healing rate to approximately 70% of wound area. PMID- 8221333 TI - [Anatomy of the skin and fascia layers of the face and advances in face lifting procedure]. PMID- 8221332 TI - [Technetium-99m bone imaging and mechanical strength test on heterogenous inorganic bone implantation]. AB - Two kinds of heterogenous inorganic bone (HAB), lower temperature treated HAB (LHAB) and higher temperature treated HAB (HHAB), were implanted and observed by technetium-99m bone imaging and mechanical strength test. The results confirmed that the LHAB possessed biocompatibility and osteoinductivity. Half a year after implantation, LHAB induced bone formation and bridged large osseous defect. PMID- 8221334 TI - [Hydrofluoric acid burns]. PMID- 8221335 TI - Harold Nathan Segall (1897-1990). PMID- 8221336 TI - Primary prevention of atherosclerosis. PMID- 8221337 TI - Canadian Consensus Conference on Thrombolytic Therapy. PMID- 8221338 TI - Coronary thrombolysis: history and rationale. PMID- 8221339 TI - Mechanisms of action of thrombolytic drugs. PMID- 8221340 TI - Rules of evidence and clinical recommendations for the management of patients. PMID- 8221341 TI - Coronary thrombolysis: mortality trials. AB - There is unequivocal evidence for the reduction of early mortality (up to about 35 days) by SK, APSAC and rt-PA, each compared with placebo, for patients treated intravenously within the first few hours of onset of acute MI. An overview of early trials demonstrating mortality reduction with SK, has been confirmed by three level I trials of SK, one level I trial of APSAC and one level I trial of rt-PA. The early benefits observed in the large trials of SK, APSAC and rt-PA have persisted at the one-year follow-up. The initial two large trials which directly compared mortalities of therapy with SK, rt-PA and APSAC detected no significant differences. However, the recently presented GUSTO trial has demonstrated greater mortality reduction with accelerated rt-PA plus heparin, compared with regimens of combined rt-PA/SK plus heparin, SK plus iv heparin, and SK plus sc heparin. PMID- 8221342 TI - Important patient subgroups in thrombolytic therapy. PMID- 8221343 TI - Coronary thrombolysis: nonmortality measures of efficacy. AB - Early reperfusion during AMI results in myocardial salvage, recovery of function and reduced mortality (level I). This can be achieved with all of the currently available thrombolytic agents (level I). Benefits from late reperfusion are not proven but there are data which suggest that delayed reperfusion is better than persistent occlusion (level II). There is now clear evidence that accelerated dose rt-PA (plus heparin) is more effective in reducing mortality than is SK (plus heparin) or a combined regimen of rt-PA/SK (plus heparin) (level I). Soon to-be published data will refine the extent to which different subgroups benefit and will provide a basis from which to select the most appropriate agent in individual patients. The overall benefit was the reduction of one death per 100 patients treated with rt-PA compared with SK, while the absolute benefit varied considerably among patient subgroups. PMID- 8221344 TI - Coronary thrombolysis: hemorrhagic complications. PMID- 8221345 TI - Nonhemorrhagic complications of coronary thrombolysis. PMID- 8221346 TI - Clinical use of coronary thrombolytic therapy. Contraindications. PMID- 8221347 TI - Coronary thrombolysis and previous SK therapy. PMID- 8221348 TI - Coronary thrombolysis: prehospital use. AB - Strong evidence supports the concept that earlier application of fibrinolytic treatment can maximize the benefits derived. The results of two level I trials have shown that prehospital thrombolysis is feasible and safe. Although the trials have not proven that prehospital thrombolysis improves prognosis, they have shown that early identification of eligible patients can greatly accelerate the process of drug administration. PMID- 8221349 TI - Coronary thrombolysis in community centres and emergency rooms. PMID- 8221350 TI - New dose regimens for coronary thrombolysis. PMID- 8221351 TI - Coronary thrombolysis: adjuvant medical therapy. PMID- 8221352 TI - Coronary thrombolysis: adjuvant coronary angioplasty. PMID- 8221353 TI - Coronary thrombolysis: adjuvant revascularization surgery. PMID- 8221354 TI - Cost-effectiveness of intravenous thrombolytic drugs for acute myocardial infarction. PMID- 8221355 TI - Canadian Consensus Conference on Coronary Thrombolysis. Recommendations. PMID- 8221356 TI - Chaos in the coronary. PMID- 8221357 TI - The dialogue between the doctor and the patient. PMID- 8221358 TI - The new science of chaos--implications for cardiology? PMID- 8221359 TI - Neuroendocrine response to cardiac transplantation. AB - OBJECTIVE: The neuroendocrine response to heart transplantation was characterized in 11 patients with special reference to long term effects on plasma hormone concentration. DESIGN: Multiple serial measurements of preload, ejection fraction, plasma renin activity (PRA), aldosterone, atrial natriuretic factor (ANF) and catecholamines were made over time. SETTING: Tertiary care cardiac referral, university-based centre. PATIENTS: Eleven adult patients undergoing orthotopic cardiac transplantation were studied. The group consisted of 10 males and one female (mean age 52 +/- 2 years). Eight patients had coronary atherosclerosis, two had idiopathic cardiomyopathy and one had valvular heart disease. All patients were in end-stage heart failure (Canadian Cardiovascular Society class IV) and two also had angina. INTERVENTIONS: Right heart catheterization and hormonal assays in blood were performed simultaneously preoperatively and postoperatively at 24 h, 48 h and during each endomyocardial biopsy. An endomyocardial biopsy to detect rejection was performed weekly for two to four weeks, then every three to four months at one year postoperatively. Hemodynamic measurements included central venous pressure (CVP) and pulmonary capillary wedge pressure (PCWP). Ejection fraction was measured in each patient using radionuclide ventriculography preoperatively and serially through the postoperative period. MAIN RESULTS: Following transplantation, transient elevation of intracardiac filling pressures occurred. The CVP and PCWP were elevated at 15 +/- 2 and 17 +/- 1 mmHg, respectively, early postoperatively (ie, days 2 to 30 postsurgery). Late postoperatively (ie, more than 30 days postoperatively), the CVP and PCWP decreased to 8 +/- 1 and 12 +/- 1 mmHg, respectively. Systolic function, as measured by radionuclide ejection fraction, did not change significantly from the early to the late postoperative period (60 +/- 5% early versus 59 +/- 2% late postoperatively). PRA and plasma aldosterone fell in association with the decrease in filling pressures (PRA was 2.4 +/- 0.8 ng/mL/h early versus 1.0 +/- 0.2 ng/ml/h late; plasma aldosterone was 122 +/- 31 pg/mL early versus 103 +/- 16 pg/mL late). Plasma aldosterone levels were similar in the early and late postoperative periods, except during the first day after surgery during which a transient elevation occurred. ANF remained markedly elevated despite the fall in filling pressures (323 +/- 50 pg/mL preoperatively, 360 +/- 33 pg/mL early postoperatively and 322 +/- 31 pg/mL late postoperatively). CONCLUSIONS: The authors conclude that transient cardiac dysfunction occurs following cardiac transplantation with elevation of filling pressures and continued increased activity of the renin-angiotensin-aldosterone system (RAAS) and elevated plasma ANF levels. With return of cardiac function and normalization of filling pressures, the activity levels of the RAAS decrease, but not those of ANF, which remain chronically abnormally elevated. It is not clear whether this persistent elevation of ANF is the result of factors related to the transplant procedure, such as extrinsic denervation or antirejection therapy, among others, or is the persistence of factors acting preoperatively. However, known interactions of cyclosporine with vascular smooth muscle and endothelial cells leading to increased sensitivity to vasopressor hormones and increased circulating levels of endothelin appear as the most likely explanation for the chronic elevation of ANF plasma levels. In this context, ANF may play a key role in moderating the side effects of cyclosporine treatment. PMID- 8221360 TI - Reversal of diastolic dysfunction by intravenous magnesium chloride. AB - OBJECTIVE: To treat diastolic dysfunction in patients with coronary artery disease (CAD). DESIGN: Open study of the intravenous injection of 5 mL of a solution containing 17% magnesium chloride (equal to 100 mg of elemental magnesium) during routine coronary angiography in patients with a raised left ventricular end-diastolic pressure (LVEDP). SETTING: A hospital department of cardiac hemodynamics. SUBJECTS: Ten patients with angiographically documented CAD. RESULTS: After magnesium chloride injection, the LVEDP was significantly reduced in all patients, from a mean of 24 +/- 3 to 16 +/- 3 mmHg (P < 0.001). CONCLUSIONS: Under the short term conditions of the study, the injection of magnesium chloride effectively improved left ventricular diastolic function. Further study of the long term effect is required. PMID- 8221361 TI - Coronary artery bypass grafting in Alberta from 1984 to 1989. AB - OBJECTIVE: To examine the geographic and temporal trends in the performance of coronary artery bypass grafting (CABG) in the province of Alberta. This examination was designed to improve understanding of the geographic distribution and temporal changes in the incidence of the procedure. DESIGN: A cohort of all individuals (2.4 million) registered with the Alberta Health Care Insurance Plan (AHCIP) was followed for the five-year period from April 1, 1984 to March 31, 1989. No new members were admitted to the cohort and an attrition rate averaging 6% per year was observed. SETTING: Data from the claims administrative database of the AHCIP were used. The AHCIP includes the records of all registered residents of the province and is virtually complete. PARTICIPANTS: Participants included all 2963 members of the cohort (2413 males, 550 females) who underwent CABG over the study period. MAIN RESULTS: The overall mean surgical rate was 28.8 per 100,000 (46.7 for males, 10.8 for females; chi 2 = 1172.8, df = 1, P < 0.0001). The mean age at surgery was 59.6 years (57.9 for males, 61.4 for females), with 70 and 76% of procedures occurring between the ages of 60 and 70 years for males and females, respectively. The largest increases in incidence were in males (223%) and females (148%) aged 70 years or older. Both males (chi 2 linearity = 25.95, df = 3, P < 0.01) and females (chi 2 linearity = 8.34, df = 3, P < 0.05) displayed an increasing trend over the study period. Data were aggregated across the five-year study period to assess geographic variations. No difference was noted across census divisions for either sex. CONCLUSIONS: The similarity in rates across census divisions implies consistent use of criteria for the performance of this procedure. Also, the results imply that research, in Alberta, should focus more on indications for the procedure and its outcomes, than on geographic disparities. PMID- 8221362 TI - Overview of clinical trials in congestive heart failure. AB - Heart failure is the only cardiovascular disorder which is increasingly prevalent in North America. Several large clinical trials have emerged within the past 10 years. The results from some of these trials have had a significant impact on the way congestive heart failure is recognized and treated. The objective of this review is not to review exhaustively all recent clinical trials in heart failure but to present the most important ones selected because of their impact on how one understands and treats congestive heart failure in 1993. The results, as well as some important pathophysiological considerations related to these trials, will be addressed. PMID- 8221363 TI - Role of endothelin in heart function in health and disease. AB - A comprehensive review of the literature has revealed that endothelins belong to a family of vasoactive peptides which are formed and released from the endothelium. By producing constriction of the coronary arteries and peripheral blood vessels, endothelins are known both to reduce coronary bloodflow and increase blood pressure and thus can be seen to affect heart function adversely. On the other hand, endothelins are capable of producing positive inotropic and chronotropic effects by directly affecting both the myocardium and nodal tissues. Prolonged actions of high concentrations of endothelins can be seen to induce relative hypoxia in the myocardium which will eventually result in heart dysfunction. The mechanisms of actions of endothelin on smooth muscle cells and cardiomyocytes include interaction with endothelin receptors on the cell surface, activation of phospholipase C through G-proteins, and increase in the intracellular concentration of Ca2+ through the increase in phosphoinositol turnover. Endothelins were found to exert no effects on sarcolemmal Na+,K(+) ATPase, Na(+)-Ca2+ exchange and Ca2+ pump systems nor on the sarcoplasmic reticular Ca2+ pump system and myofibrillar ATPase activities in the rat heart. Marked elevation in the levels of plasma endothelins and down-regulation of endothelin receptors in ischemia-reperfusion injury, hypertension and chronic diabetes indicate a significant role of endothelins in the genesis of heart dysfunction under different pathological conditions. PMID- 8221364 TI - Repeated plaque rupture in the same bypass graft. AB - A 51-year-old man experienced a plaque rupture at the site of a shallow plaque in his anterior descending bypass graft and went on to suffer a myocardial infarct. Angiography, five months later, showed excellent recanalization; however, one month after that angiogram, the patient presented with unstable angina and evidence of plaque rupture at the same site. The presence of plaque in a graft indicates risk for future plaque rupture, but there appear to be no definite indicators to identify the specific plaque that is at highest likelihood of rupture. PMID- 8221365 TI - Patterns of recurrence and survival in AMI patients in Canada. AB - OBJECTIVE: To describe the patterns of recurrence and survival of patients following their first acute myocardial infarction (AMI). DESIGN: A retrospective follow-up study based on the population of Nova Scotia and Saskatchewan from 1981 85. SUBJECTS: The analyses were performed on patients aged 25 to 75 years (n = 2083) who had been admitted to hospital with their first AMI during 1981 from the provinces of Nova Scotia and Saskatchewan. MAIN RESULTS: The rate of recurrence of AMI was 10% in men and 7% in women, regardless of age. Median time to recurrence was 13 to 15 months in men and seven months in women. After the first AMI, mortality rates were higher in women one week and one month after the initial infarction, reaching statistical significance (only in the 55- to 74-year old age group)--P < 0.01 and P < 0.02, respectively. Five-year mortality rates were significantly higher in the older age groups (P < 0.01), but were similar in men and women. CONCLUSIONS: Reinfarction occurs more quickly in women, who also appear to be at a higher risk of death during the first month, and especially during the first week, postinfarction. PMID- 8221366 TI - Comparison of the oculokinetic perimetry glaucoma screener with two types of visual field analyser. AB - We compared the accuracy and testing time of a new 26-point central visual field glaucoma screening device, the oculokinetic perimetry (OKP) screener, with the results obtained with the Friedmann field analyser standard program (phase 1) and the Humphrey field analyser 40-point screening program (phase 2). Seventy-three eyes of 73 subjects were tested in phase 1, and 142 eyes of 75 subjects in phase 2. The study population comprised patients with eyes that were ocularly normal and patients with eyes showing early cataract, posterior vitreous detachment, established chronic open-angle glaucoma or ocular hypertension. The OKP glaucoma screener demonstrated a sector concordance rate of 95% in phase 1 and 93% in phase 2. The OKP screener was three to four times more rapid in execution than the standard device. We feel that the OKP glaucoma screener is ideally suited for use by physicians in a family practice setting. PMID- 8221367 TI - Recession of the superior oblique tendon for inferior oblique palsy and Brown's syndrome. AB - Superior oblique tendon recession of 12 to 14 mm was performed in five patients with inferior oblique muscle palsy and three patients with Brown's syndrome. The angle of deviation was decreased by an average of 18 prism dioptres in the worst field of gaze for the former group and by 17.3 delta for the latter group. As a secondary benefit of the procedure for the patients with inferior oblique palsy, any associated preoperative A-pattern was collapsed an average of 10 delta postoperatively. The procedure eliminated all abnormal head postures and improved versions and ductions for both groups. Symptoms of diplopia and torsion were resolved, and full fusional ability was maintained at preoperative levels in all fields of gaze for all patients. After an average follow-up period of 18 months none of the patients had manifested superior oblique palsy or other notable postoperative surgical complications. Theoretical advantages of superior oblique tendon recession include the potential for reversibility and reoperation if required, the possibility of asymmetric surgery and the potential to perform the procedure with an adjustable suture. The technique is an advantageous approach in the surgical treatment of these complicated cases. PMID- 8221368 TI - Brown's syndrome with contralateral inferior oblique overaction: a possible mechanism. AB - The simultaneous occurrence of Brown's syndrome in one eye and inferior oblique overaction in the other eye is an infrequently reported association. Review of our series of 38 cases of Brown's syndrome disclosed six patients with overaction of the contralateral inferior oblique, of whom four stepped out to contralateral superior oblique palsy. We propose that these patients initially have bilateral Brown's syndrome in infancy, and spontaneous resolution occurs in one eye only. While Brown's syndrome is present the antagonist inferior oblique muscle undergoes isometric contracture. With spontaneous resolution of Brown's syndrome a relative imbalance of forces occurs, with the superior oblique muscle now being relatively paretic compared with the contracted and fibrotic inferior oblique. Up shoot in adduction then becomes apparent. PMID- 8221369 TI - Unexpected stereoacuity following surgical correction of long-standing horizontal strabismus. AB - Eight patients with long-standing large-angle constant tropias unexpectedly achieved stereoacuity (40 seconds of arc in six patients and 60 seconds of arc in two patients) following strabismus surgery. There are few reports in the literature documenting this finding. The authors found that if excellent postoperative alignment can be achieved in patients with good bilateral visual acuity, some of these patients will demonstrate high-grade stereoacuity even if the preoperative findings suggest this outcome to be unlikely. PMID- 8221370 TI - Presumed photic retinopathy after cataract surgery: an angiographic study. AB - To ascertain the prevalence rate of presumed photic retinopathy after cataract surgery, 276 fluorescein angiograms of 147 patients were reviewed. The diagnosis was based on strict angiographic criteria. Lesions consistent with healed photic retinopathy were noted in four patients (prevalence rate of 3%). This finding is discussed and comparisons with the literature are made. PMID- 8221371 TI - Pars plana vitrectomy in the management of vitreous touch syndrome. AB - The management of three patients with aphakic bullous keratopathy by means of pars plana vitrectomy is described. The patients had previously undergone uneventful intracapsular cataract extraction with intact vitreous face. Eventual herniation of the vitreous into the anterior chamber with resultant corneal endothelial touch caused the corneal edema in all three cases. Pars plana vitrectomy resulted in resolution of photophobia and irritation. The visual acuity improved from 20/200, 20/200 and 20/70 to 20/70, 20/60 and 20/40 respectively. The central cornea was cleared of edema in all three cases. The final vision was limited in two of the patients by the persistence of preoperative cystoid macular edema. Pars plana vitrectomy may be used in the management of selected cases of aphakic bullous keratopathy from vitreous touch. PMID- 8221372 TI - Mucoepidermoid carcinoma of the lacrimal sac: a report of two cases, with observations on the histogenesis. PMID- 8221373 TI - Management of severe hypotony with intravitreal injection of Healon. PMID- 8221374 TI - Metastatic group B streptococcal endophthalmitis from a cutaneous foot ulcer. PMID- 8221375 TI - Macular infarction after intravitreal injection of amikacin. PMID- 8221376 TI - Complications of retinotomies for subretinal fluid drainage. PMID- 8221377 TI - Sequence analysis of the lysin gene region of the prolate lactococcal bacteriophage c2. AB - Approximately 80% of the genome of the prolate-headed lactococcal bacteriophage c2 was cloned into shuttle vectors pSA3 and pFX3 in Escherichia coli and transferred to Lactococcus lactis. A 1.67-kilobase EcoRV fragment containing the gene for the phage lysin was identified and the position and orientation of the phage lysin gene in the physical map of the phage were determined. The phage lysin was expressed in E. coli and its sequence was determined and compared with the sequences of other bacteriophage lytic genes. The sequence was similar, but not identical, to that of the related lactococcal phage m13, having a number of silent substitutions and an apparent deletion that altered the carboxy terminus of the protein. Possible alternative translation initiation codons for the lysin gene and two possible alternative mechanisms for access of the lysin enzyme to the cell wall are discussed. An open reading frame upstream of the putative lysin gene was found to be 177 base pairs longer than that reported for phage m13. A codon usage table for the lysin genes of several phages as well as for reported gene sequences from L. lactis and lactococcal bacteriophages is presented. PMID- 8221378 TI - Digestion of cell-wall monosaccharides of ryegrass and alfalfa hays by the ruminal bacteria Fibrobacter succinogenes and Butyrivibrio fibrisolvens. AB - The ruminal bacteria Fibrobacter succinogenes strains S85 and BL2 were grown in monocultures or in coculture with strain D1 of Butyrivibrio fibrisolvens, and the solubilization of ryegrass and alfalfa cell walls (CW) and digestion of CW monosaccharides were measured. Fibrobacter succinogenes monocultures and cocultures with B. fibrisolvens D1 degraded 58-69% of ryegrass CW, solubilizing 67-78% of CW glucose, 65-71% of CW xylose, 69-75% of hemicellulose, and 68-77% of total CW monosaccharides. When grown on alfalfa CW, those cultures degraded 28 39% of alfalfa CW, solubilizing 42-58% of CW glucose, 30-36% of CW xylose, and 37 45% of hemicellulose. With respect to both substrates, F. succinogenes strains solubilized CW carbohydrates better than did B. fibrisolvens D1. Complementary interaction between B. fibrisolvens D1 and the F. succinogenes strains was identified with respect to the utilization of some solubilized carbohydrates, but not with respect to the extent of CW solubilization, which was determined mainly by the F. succinogenes strains. For both substrates, utilization of cellulose by F. succinogenes monocultures was high (96-98%), whereas that of hemicellulose was lower (24-26% in ryegrass and 49-50% in alfalfa). Under scanning electron microscopy, F. succinogenes bacterial cells attached to and colonized on CW particles were characterized by the appearance of protuberant surface structures that we have identified as "polycellulosome complexes." PMID- 8221379 TI - Activity of microbial peptidases, oxidases, and esterases in lake waters of varying trophic status. AB - The range and activities of microbial enzymes present in lake water were assessed directly in cells concentrated by tangential flow filtration. A total of 108 enzymes were assayed in this study, which included tests for 60 peptidases, 20 oxidases, and 10 esterases, and 18 miscellaneous tests. In general, no trends in the range of enzymes were associated with trophic status of the lakes. However, one lake that was hypereutrophic had a greater range of enzymes than the other lakes tested. An increase in total enzyme activity (activity/mL) was recorded with an increase in trophic status of the water. The relationship between the physical and chemical attributes of each lake and microbial enzyme activities was investigated by principal component analysis. Quantitative changes between lakes in 11 of the 21 variables were shown to be closely related to changes in the enzyme activities of the lakes; total organic carbon, particulate carbon, particulate nitrogen, pH, and chlorophyll a showed the closest relationships. PMID- 8221380 TI - Physiology of basal ganglia disorders: an overview. AB - The pathophysiology of the movement disorders arising from basal ganglia disorders has been uncertain, in part because of a lack of a good theory of how the basal ganglia contribute to normal voluntary movement. An hypothesis for basal ganglia function is proposed here based on recent advances in anatomy and physiology. Briefly, the model proposes that the purpose of the basal ganglia circuits is to select and inhibit specific motor synergies to carry out a desired action. The direct pathway is to select and the indirect pathway is to inhibit these synergies. The clinical and physiological features of Parkinson's disease, L-DOPA dyskinesias. Huntington's disease, dystonia and tic are reviewed. An explanation of these features is put forward based upon the model. PMID- 8221381 TI - Pathogenesis of mesial temporal sclerosis. AB - A relationship between epilepsy and damage to mesial temporal structures has long been recognized. Recent advances have clarified somewhat the issue of whether the pathological changes seen in mesial temporal sclerosis represent the cause or the effect of seizures. This paper reviews mesial temporal sclerosis from an historical perspective and summarizes recent developments in the fields of excitotoxicity, selective vulnerability, and synaptic reorganization as they pertain to the pathogenesis of mesial temporal sclerosis. PMID- 8221382 TI - Nuclear magnetic resonance study of cerebrospinal fluid from patients with multiple sclerosis. AB - Proton nuclear magnetic resonance (NMR) spectroscopy was used to examine cerebrospinal fluid (CSF) from patients (n = 30) with actively progressive multiple sclerosis (MS). Metabolite concentrations obtained from the spectra were compared to those determined from the spectra of CSF from control patients (n = 27) with benign spinal disorders. No significant difference was found between the 2 groups for most constituents, including lactate, glutamine, citrate, creatine and creatinine, and glucose. Acetate levels were significantly higher in MS patients, while formate levels were significantly lower, than the controls. There were no significant differences in metabolite concentrations in CSF from early and longstanding MS patients. A peak due to an unidentified compound was found at 2.82 ppm in the spectra of CSF from patients with actively progressive MS, but not in the spectra of CSF from the controls. The peak was not found in spectra of CSF from patients with AIDS dementia complex (n = 9) or Parkinson's disease (n = 5), but it did appear in spectra of CSF from 1 patient with Jakob-Creutzfeldt disease (out of 3 examined) and from 1 patient (out of 7) with Guillain-Barre disease. The unidentified compound is volatile and, from the chemical shift of the observed NMR peak, is probably an N-methyl compound. As such, it may be an intermediate in the cholinoglycine cycle, in which an abnormality has been proposed to exist in MS patients. PMID- 8221383 TI - Severe permanent encephalopathy in acute lymphoblastic leukemia. AB - As survival rates for childhood acute lymphoblastic leukemia have increased, concerns over improved quality-of-life have also increased. Although 3-10% of children may experience acute transient neurotoxicity during induction chemotherapy, they are felt to be at low risk for late sequelae. We report three previously healthy boys with newly-diagnosed acute lymphoblastic leukemia who presented with obtundation and severe seizures during late induction with a standard four drug chemotherapy regimen. While all three are disease-free survivors, they unexpectedly have persistent and medically intractable partial complex seizures, broad-based neuropsychological impairment and striking neuroimaging abnormalities. These findings suggest that children with leukemia who develop an acute encephalopathy during induction chemotherapy are at risk for long-term neurological and neuropsychological sequelae, despite the cessation of further potentially neurotoxic therapy. PMID- 8221384 TI - Serum lipoprotein fatty acid profile in hereditary ataxias. AB - We investigated the serum fatty acid profiles of cholesterol esters, phospholipids and triglycerides in 24 patients with Friedreich's disease and 16 patients with other forms of spinocerebellar degeneration. In 8 patients with Friedreich's disease we also analyzed the fatty acid profile of the lipoprotein fractions. We found no major differences in fatty acid profiles between ataxic patients and sex and age-matched controls; in particular there was no decrease of linoleic acid in Friedreich's disease. The level of linoleic acid in serum cholesterol esters decreased with increasing disability of patients. PMID- 8221385 TI - The usefulness of CT scanning in diagnosing dementia of the Alzheimer type. AB - In order to determine the usefulness of Computerized Tomography (CT) scanning in making a diagnosis of dementia of the Alzheimer type, a group of patients diagnosed by NINCDS-ADRDA criteria (n = 22) were compared to a group of normal subjects (n = 49) using certain defined linear CT scan measurements. These measurements included specific measurements of the temporal lobes (temporal horns). Subjects were classified correctly 91.5% of the time with a high degree of probability. A diagnostic equation is presented which will allow testing of these methods in a prospective fashion. PMID- 8221386 TI - The use of antiepileptic drug levels in children: a survey of Canadian pediatric neurologists. AB - There are 60 pediatric neurologists in Canada. Replies were received from 56 in response to a survey regarding the use and perceived value of antiepileptic drug (AED) levels. AED levels are frequently ordered and influence clinical care. There were, however, discrepancies among pediatric neurologists regarding the upper and lower limits of the "therapeutic ranges" and the clinical application of levels. We suggest that both the value and use of AED levels needs further study. PMID- 8221387 TI - Contact thermography in the diagnosis of childhood migraine. AB - The objective of our prospective study was to assess the role of contact thermography in children with migraine. Contact thermograms were done in 54 children aged 4.2-16.5 years (median 10.5 years), who were seen for headache and on 10 age-matched controls, between July and December 1991. Thermograms were interpreted as definitely normal, equivocally normal, equivocally abnormal, and definitely abnormal by a radiologist who was blinded to clinical information. Forty-eight children had the test between headaches; of these, four out of 26 patients (15%) who had migraine without aura and 3 out of 14 children (21%) who had migraine with aura had definitely abnormal thermograms. Nine out of 10 normal controls had definitely normal thermograms. The sensitivity of contact thermography in the diagnosis of childhood migraine, when done between headaches, was low in our study. PMID- 8221388 TI - Inflammatory myopathy and Walker-Warburg syndrome: etiologic implications. AB - Walker-Warburg syndrome is a well delineated clinical entity with characteristic brain and eye anomalies. Recent diagnostic surveys have revealed that muscular dystrophy is an obligatory feature of this syndrome. We report a patient with an inflammatory myopathy that preceded dystrophic changes. While reports of parental consanguinity and multiple affected sibships strongly suggest an autosomal recessive genetic basis for this syndrome, previous pathological analyses of the CNS have suggested an inflammatory process. Our case supports both the notion of an aberrant inflammatory process that is likely under genetic control or etiologic heterogeneity (phenocopies) underlying this syndrome. PMID- 8221389 TI - Dural cavernous hemangioma: an under-recognized lesion mimicking meningioma. AB - We report a 77-year-old woman who presented with partial seizures and was found to have an enhancing dural-based parietal convexity mass. The lesion enlarged on serial examination by computed tomography (CT) over a one year period. The clinical features and radiologic appearance were compatible with a pre-operative diagnosis of meningioma; however, pathologic findings were typical of a dural cavernous hemangioma. Accumulating evidence suggests that these lesions are an uncommon but distinct type of vascular malformation most often arising from the cavernous sinus, tentorium, or cerebello-pontine angle. With CT, magnetic resonance imaging and angiography, these lesions can closely resemble meningioma in terms of signal characteristics, enhancement pattern, and location. This is of importance both in the practical management of meningiomas where the diagnosis is often based on radiologic studies alone, and in clinical trials where incorrect entry diagnosis should be avoided. PMID- 8221390 TI - Failure to recognize status epilepticus in a paralysed patient. AB - Paralysis induced by neuromuscular blocking agents facilitates ventilation of seriously ill patients but may preclude clinical recognition of seizures. We describe the occurrence of severe cognitive impairment in a 14-year-old girl in whom status epilepticus was recognized only when pancuronium was withdrawn after 14 hours of paralysis. This patient emphasizes a potential danger of paralysis from drugs in patients with acute cerebral dysfunction. PMID- 8221391 TI - Tremorgenic encephalopathy: a role of mycotoxins in the production of CNS disease in humans? AB - We describe a young man who, shortly after exposure to moldy silage, developed a neurological syndrome consisting of dementia and a remarkable tremor which was precipitated by movement. All symptoms resolved within one week. Despite investigation, no definitive diagnosis was reached. We propose that this patient's illness may have resulted from inhalation exposure to a tremorgenic mycotoxin. PMID- 8221392 TI - Syrinx extending from conus medullaris to basal ganglia: a clinical, radiological, and pathological correlation. AB - A 41-year-old woman with a history of birth injury to the brachial plexus suffered several delayed episodes of neurological deterioration. Magnetic resonance imaging studies revealed a syrinx extending from the conus medullaris into the brainstem and rostrally into both internal capsules. She died of an acute exacerbation of chronic respiratory failure. Autopsy demonstrated syringomyelia and syringobulbia with cavity extension bilaterally along the corticospinal tracts into the internal capsules. Islands of glial tissue in the subarachnoid space around the medulla caused obstruction of the subarachnoid space at the foramen magnum. These were probably the result of birth injury to the cerebellum. A detailed clinico-pathological correlation is provided to explain her neurological deficits. The pathogenesis of syrinx formation is discussed in terms of a late manifestation of birth trauma. PMID- 8221393 TI - Concerning mind. AB - The nature of the human mind is addressed from a neurological viewpoint. First from reported observations on subjects born blind with congenital cataract, who acquired vision after surgery, the concept of the primacy of somatosensation is developed. Therefrom some principles of the organization of higher sensory functions in the dominant parietal, occipital and temporal lobes are deduced. Sentience is traced to somatosensation. Some characteristics of the mind and of thinking are described. The non-existence of the unconscious is inferred. The question of free will is approached. Using neurological experience it is demonstrated that a good case for operation of a free will is difficult to establish. Positing the absence of a free will may be beneficial in the analysis and management of behavior. Finally the opportunity to study the nature and mechanism of the emotions in clinical neurology is outlined. PMID- 8221394 TI - William John Horsey, M.D., F.R.C.S.C. Neurosurgeon to St. Michael's Hospital, Toronto. PMID- 8221395 TI - Elemental iodine: relief for the painful breast? PMID- 8221396 TI - Medical memories: an aortoesophageal fistula. PMID- 8221397 TI - The influence of intermittent positive-pressure ventilation on the cardiorespiratory dynamics of diaphragmatic rupture with gastric herniation. AB - To determine the effect of intermittent positive-pressure ventilation (IPPV) on gastric herniation in diaphragmatic rupture, 16 piglets underwent laceration of the left hemidiaphragm. Arterial blood gas levels, pulmonary artery wedge pressure, cardiac output and arterial blood pressure were measured. The stomach was then placed above the diaphragm. The initial measurements were repeated, and displacement of the stomach above the diaphragm (S/D) was measured. The animals were divided into two groups: group 1, eight animals breathing 80% oxygen spontaneously and group 2, eight animals receiving IPPV. Initially in group 2 there was only a small decrease in mean (+/- SD) arterial oxygen tension from 484 +/- 34 mm Hg to 424 +/- 20 mm Hg (other parameters were unchanged). In group 1, gastric herniation produced a further fall in arterial oxygen tension to 308 +/- 10 mm Hg at 1 hour, a rise in arterial carbon dioxide tension to 49 +/- 4 mm Hg and a decrease in pH to 7.32 +/- 0.04. In group 2 similar changes in arterial blood gas levels occurred with gastric herniation, but there was a return to baseline values of 490 +/- 28 mm Hg for arterial oxygen tension, 37 +/- 4 mm Hg for carbon dioxide tension and 7.38 +/- 0.05 for pH after IPPV. Similar changes were seen in blood pressure, cardiac output and pulmonary artery wedge pressure. S/D remained at 6.3 +/- 0.2 cm in group 1 but decreased from 6.0 +/- 0.2 cm to 1.1 +/- 0.5 cm after 1 hour in group 2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221398 TI - Effect of cardiac stabbing on ventricular function: evaluation by radionuclide angiography. AB - OBJECTIVE: To determine whether cardiac stab wounds cause early ventricular dysfunction. DESIGN: Retrospective case series over 7 years. SETTING: University affiliated teaching hospital. PATIENTS: Nine consecutive survivors of cardiac stab wounds. INTERVENTIONS: Postoperative radionuclide angiography performed in all patients between 1 and 13 days after injury. MAIN OUTCOME MEASURES: Information on the patient's clinical presentation and hospital course was obtained from the patient's charts and correlated with ventricular function as measured by the right and left ventricular ejection fractions (RVEF and LVEF). RESULTS: Cardiac injuries consisted of isolated right (five) and left (three) ventricular lacerations, and one biventricular laceration. Three patients had perioperative dysrhythmias (two ventricular, one atrial), and nonspecific electrocardiographic changes occurred in seven patients. Six patients had postoperative complications: pericarditis in three and pneumonia, pulmonary thromboembolism and abdominal wound dehiscence in one each. The LVEF was normal in all patients. Although none had clinically apparent cardiac dysfunction at the time of radionuclide angiography, six patients had a reduced RVEF (less than 0.45). All patients remained asymptomatic at the time of discharge from hospital (median 12 days postoperatively). In two of the six patients who had a reduced RVEF, one had a normal RVEF 2 months later; the other had a near-normal RVEF (0.43) but had persistent right ventricular enlargement 7 months later. Both were asymptomatic. CONCLUSIONS: Although a reduced RVEF is common in survivors of cardiac stab wounds, the reduction is small and is not associated with clinically apparent cardiac dysfunction. PMID- 8221399 TI - Recycling of cardiopulmonary bypass cannulae: a risk-benefit analysis. AB - OBJECTIVE: To determine whether recycling of arterial and venous cannulae for cardiopulmonary bypass is associated with an increased risk of infectious complications after open heart surgery. DESIGN: A prospective, risk-benefit analysis. SETTING: A university teaching hospital. PATIENTS: Five hundred and forty patients underwent a variety of non-transplant cardiac operations, which were performed by one surgeon between October 1988 and July 1993. INTERVENTIONS: Standard techniques of open heart surgery were used. Ascending aortic and vena caval cannulae and the multiple perfusion device for administering cardioplegia down saphenous vein bypass grafts were reused an average of 10 to 15 times after sterilization with ethylene oxide after each use. MAIN OUTCOME MEASURES: The postoperative morbidity and mortality and the estimated savings achieved by recycling the cannulae. RESULTS: The operative mortality (total in hospital plus within 30 days of operation) was 2.8% (15 patients). There were no cases of endocarditis; there was only one case of superficial presternal infection and one case of mediastinitis. Gram-negative organisms were cultured from the blood of two patients and gram-positive organisms from the blood of one patient. Only one patient died of an infectious complication. The estimated cost saving during the study period was more than $50,000. CONCLUSIONS: The risk of wound and hematogenous infection after open heart surgery in which recycled cannulae are used is less than 1%. This rate is lower than that quoted in most recent reports. Reuse of heart-lung bypass cannulae may realize substantial cost savings without detriment to the patient. PMID- 8221400 TI - Myocardial and circulatory performance during the ischemic phase of superior mesenteric artery occlusion. AB - OBJECTIVE: To investigate myocardial and circulatory parameters during the acute ischemic phase of mesenteric artery occlusion. DESIGN: A prospective, randomized, control trial. SUBJECTS: Twelve, adult, mongrel dogs. INTERVENTIONS: In seven dogs the superior mesenteric artery was occluded with two silk ligatures (experimental group). In five dogs the ligatures were not tied (control group). Measurements were made during 7 hours of occlusion. MAIN OUTCOME MEASURES: Myocardial performance and circulatory performance. RESULTS: There were significant (p < 0.05) reductions in arterial blood pressure, mean pulmonary artery pressure, diastolic pulmonary artery pressure and cardiac index in dogs exposed to intestinal ischemia compared with the control dogs. No differences were identified in ventricular performance, stroke volume index or peripheral vascular resistance index. CONCLUSIONS: Early cardiac and central circulatory changes in massive intestinal ischemia are due to intravascular hypovolemia. Sepsis and myocardial depressant factors were not found to be a cause of death. PMID- 8221401 TI - Reconstruction of composite facial defects: the combined application of multiple reconstructive modalities. AB - OBJECTIVE: To describe the combined use of craniofacial skeletal reconstruction, tissue expansion and microvascular free tissue transfer in the repair of major composite facial defects. DESIGN: Case series with an integrated team approach. SETTING: Craniofacial unit, university teaching hospital. PATIENTS: Three cases were selected to best illustrate the combined use of the three modalities in reconstruction of acute traumatic, congenital and post-traumatic facial defects. A 15-year-old boy had a shotgun wound to the face; a 23-year-old man had Treacher Collins syndrome; and a 55-year-old woman had a post-traumatic composite defect of the central midface. INTERVENTIONS: Preoperatively, complete neurologic, ophthalmologic and dental examinations, anthropometric analysis, prosthodontic assessment, computed tomography and computer graphics. Operatively, craniofacial exposure followed established surgical principles. Skeletal reconstruction was performed to provide accurate positioning of bony segments and three-dimensional stability. Bone grafting was used when necessary to restore bony continuity and increase stability. Tissue expansion was used to provide more locally available tissue for wound closure and resurfacing composite defects. Microsurgical free tissue transfer was used to provide functional replacement of deficient tissues. RESULTS: The results of the modalities used for reconstruction of these composite facial defects are illustrated for each case described. CONCLUSIONS: The use of multiple modalities, including craniofacial skeletal reconstruction, tissue expansion and free tissue transfer, allow the surgeon to address the specific functional and anatomical requirements associated with composite facial defects that are characterized by a combined deficiency of multiple tissues in the craniofacial region. PMID- 8221402 TI - Iodine replacement in fibrocystic disease of the breast. AB - OBJECTIVE: To determine the response of patients with fibrocystic breast disease to iodine replacement therapy. DESIGN: Review of three clinical studies beginning in 1975: an uncontrolled study with sodium iodide and protein-bound iodide; a prospective, control, crossover study from iodide to molecular iodine; and a prospective, control, double-blind study with molecular iodine. SETTING: University affiliated breast-treatment clinics. PATIENTS: Study 1: 233 volunteers received sodium iodide for 2 years and 588 received protein-bound iodide for 5 years. Study 2: the treatment of 145 patients from study 1 treated with protein bound iodide for several months who still had symptoms was switched to molecular iodine 0.08 mg/kg; 108 volunteers were treated initially with molecular iodine. Study 3: 23 patients received molecular iodine, 0.07 to 0.09 mg/kg body weight; 33 received an aqueous mixture of brown vegetable dye and quinine. The numbers in study 2 increased over the review period so that 1365 volunteers were being treated with molecular iodine by 1989. INTERVENTIONS: All patients in study 3 had pre- and post-treatment mammography and measurement of serum triiodothyronine, thyroxine and thyroid-stimulating hormone levels. MAIN OUTCOME MEASURES: Subjective evaluation--freedom from pain--and objective evaluation--resolution of fibrosis. RESULTS: Study 1: 70% of subjects treated with sodium iodide had clinical improvement in their breast disease, but the rate of side effects was high; 40% of patients treated with protein-bound iodide had clinical improvement. Study 2: 74% of patients in the crossover series had clinical improvement, and objective improvement was noted in 72% of those who received molecular iodine initially. Study 3: in the treatment group 65% had subjective and objective improvement; in the control group there was a subjective placebo effect in 33% and an objective deterioration of 3%. CONCLUSIONS: The fibrocystic breast reacts differently to sodium iodide, protein-bound iodide and molecular iodine. Molecular iodine is nonthyrotropic and was the most beneficial. PMID- 8221403 TI - [Patellectomy: results of a long-term follow-up]. AB - The authors are in disagreement with the literature on the results of patellectomy after comminuted fracture of the patella. They assessed retrospectively 17 patients who underwent unilateral total patellectomy between 1955 and 1980. Subjective and objective (clinical examination, radiologic examination and Cibex II studies) assessment was carried out for each patient using the contralateral healthy knee as a control. The results demonstrated 88% patient satisfaction, no ligamentous instability and no early arthrosis but a decrease in extensor muscle strength to 75% of normal. The authors conclude that total patellectomy is better tolerated and causes less morbidity than suggested in the literature. PMID- 8221404 TI - Intramedullary nailing of femoral shaft fractures: reoperation and return to work. AB - OBJECTIVE: To define the functional outcome and residual disability in adults treated for isolated femoral shaft fractures. DESIGN: A retrospective case study of HMRI records. The mean follow-up was 34.5 months from the time of injury. SETTING: A tertiary-care teaching hospital. PATIENTS: One hundred and thirty adults with femoral shaft fractures. Of these, 47 were isolated and qualified for study. INTERVENTIONS: Primary reamed intramedullary nailing performed between April 1987 and September 1990. MAIN OUTCOME MEASURES: Reoperation and return to work. RESULTS: Of the 47 patients, primary healing occurred in 41 (87%) patients; 6 (13%) patients had delayed union or nonunion. Twenty (43%) patients had delayed implant-related pain after fracture healing; 85% of these had relief of pain after implant removal. Of 41 patients available for analysis of function based on return to work, 33 (80%) had returned to work full time in their original occupation at the time of follow-up, and 4 (10%) were working in other full-time employment; 3 (7%) were working part time. Only one (2%) patient was disabled and unable to work. CONCLUSIONS: Primary reamed intramedullary nailing is an effective method of achieving consistent primary union in adults with femoral shaft fractures. However, the majority of patients will require secondary surgery. Most patients will be able to return to work after these injuries. PMID- 8221405 TI - Anterior and posterior spinal fusion: comparison of one-stage and two-stage procedures. AB - OBJECTIVE: To compare postoperative morbidity and length of hospital stay after combined anterior and posterior spinal fusion for patients treated by a one-stage procedure and those treated in two stages, 1 to 2 weeks apart. DESIGN: Retrospective review of all cases handled by the first author between July 1989 and November 1991. The patients had been referred for treatment of severe spinal deformity, scoliosis or kyphosis by combined anterior and posterior spinal fusion. SETTING: The operations were performed at McMaster University Medical Centre, Hamilton, Ont., by the first author, who was assisted by another specialist in spinal surgery and a fellow or resident. PATIENTS: Eleven operations of each type were performed. The mean age of the 22 patients was 16.6 and 14.6 years for those who underwent the one-stage and the two-stage procedures respectively. The diagnoses included neuromuscular disease, neurofibromatosis, spina bifida, congenital kyphoscoliosis and severe idiopathic spinal curvature. The one-stage procedure was used after the surgical team became able to provide the care associated with this type of major surgery; selection of patients also involved preoperative risk assessment and the feasibility of combining two surgical procedures that would take a maximum of 9 hours. The preoperative plan was to spend a maximum of 9 hours in performing the one-stage procedure. INTERVENTIONS: Similar surgical procedures were performed in both groups. The average number of intervertebral levels fused during the anterior component of the operation was 4.6 in the one-stage procedure and 6.0 in the two-stage procedure. Thoracolaparotomy was performed in four patients who underwent the two stage procedure. During the posterior component of the operation, instrumentation was inserted through an average of 11.6 and 12.6 intervertebral levels in the groups undergoing the one-stage and the two-stage procedures respectively. Total operating time averaged 7 hours and 15 minutes for the one-stage procedure and 11 hours for the two-stage procedure. Mean blood loss was 1830 mL for the one-stage procedure and 2270 mL for the two-stage procedure. MAIN OUTCOME MEASURES: The number of days spent in the intensive care unit (ICU) and the total number of days spent in hospital, as well as morbidity after the operation, were determined. RESULTS: Postoperative morbidity included five minor complications in patients who underwent a one-stage procedure; all of these complications resolved well. Among those who underwent the two-stage procedure there were 11 complications; in two patients further surgery was required. The patients' stay in the ICU averaged 2.6 and 7.7 days respectively for one-stage and two-stage procedures, and the total stay in hospital averaged 14 and 33 days respectively. CONCLUSIONS: When possible, the one-stage procedure for anterior and posterior spinal fusion is preferred over the two-stage procedure because of a significant reduction in the length of stay in the ICU and in hospital, as well as reduced morbidity. However, this analysis should be interpreted cautiously because of the small number of cases and the variables encountered in treating this type of spinal deformity. PMID- 8221406 TI - Bochdalek hernia with hemorrhage in an adult. AB - Life-threatening herniation of intra-abdominal viscera through a patent Bochdalek foramen is well recognized in neonates. Persistent foramina, leading to clinical problems in adult life, are very rare. In the case of the 17-year-old girl described in this paper, two-thirds of the stomach had inverted and passed into the left side of the chest as had the splenic flexure of the colon and most of the greater omentum. There was 500 mL of blood free in the peritoneal cavity and 1200 mL in the left pleural cavity, with no hernial sac. The defect measured 5 cm in diameter. It was repaired primarily after reduction of the viscera. Prompt operative intervention is recommended in such cases to prevent strangulation and bleeding from engorged viscera. PMID- 8221407 TI - Aorto-esophageal fistula secondary to benign Barrett's ulcer: a rare cause of massive gastrointestinal hemorrhage. AB - Aorto-intestinal fistula is a rare cause of massive upper gastrointestinal bleeding. The authors report on an 82-year-old man who had an aorto-esophageal fistula due to a benign Barrett's ulcer. The patient presented with a sentinel hemorrhage followed by fatal exsanguination despite vigorous attempts at resuscitation. Management of the entity requires awareness, appropriate radiologic and endoscopic investigation and early surgery. Even with appropriate management, the mortality remains high. PMID- 8221408 TI - The anatomical basis for laparoscopic splenectomy. AB - The anatomy of the splenic blood supply is extremely variable. Two principal patterns, the distributed and the magistral types, include the majority of variations. These two types differ with respect to the length of the splenic trunk, the number of arterial branches and the proportion of the medial surface of the spleen occupied by the branches as they enter the organ. The suspensory ligaments of the spleen, including the sustentaculum lienis, are usually avascular except for the gastrocolic and lienorenal ligaments on the medial side. The tail of the pancreas touches the spleen in 30% of cases and lies within 1 cm of the spleen in 73%. Knowledge of these basic anatomic facts and the variations should contribute to the safety of laparoscopic splenectomy. PMID- 8221409 TI - Ethics and health care costs. PMID- 8221410 TI - Ethics and health care costs. PMID- 8221411 TI - Ethics and health care costs. PMID- 8221412 TI - New Zealand and Canadian health care systems. PMID- 8221413 TI - Health care research. PMID- 8221414 TI - Canadian National Breast Screening Study: response. PMID- 8221415 TI - Objective-structured clinical examination. PMID- 8221416 TI - Diagnostic testing in chronic urticaria and angioedema. PMID- 8221417 TI - Accredited mammography units. PMID- 8221418 TI - Ambulatory care training. PMID- 8221419 TI - Traditional Chinese medicine. PMID- 8221420 TI - Literacy and health. PMID- 8221421 TI - Carbon monoxide poisoning. PMID- 8221422 TI - Physicians and the pharmaceutical industry. PMID- 8221423 TI - Unravelling genes: history, health care and the techno-industrial imperative. PMID- 8221424 TI - Interactions between physicians and the pharmaceutical industry: what does the literature say? AB - OBJECTIVE: To determine the effect of three types of interaction between physicians and the pharmaceutical industry--company-funded clinical trials, company-sponsored continuing medical education (CME) and information for physicians supplied by pharmaceutical detailers--on orientation and quality of clinical trials, content of CME courses and physicians' prescribing behaviour. DATA SOURCES: MEDLINE and HEALTH searches for English-language articles published from 1978 to 1993, supplemented by material from the author's personal collection. STUDY SELECTION: A total of 227 papers from the MEDLINE and HEALTH searches and about 2000 items from the author's library were initially reviewed. The following selection criteria were used: studies conducted in Australia, Canada, New Zealand, Britain and the United States; studies conducted after 1977; quantitative surveys containing details of the survey methods; studies on the orientation and quality of company-funded clinical trials and on the content of CME courses giving explicit criteria used in the evaluation; and reports on the outcome of interactions stating how the outcomes were assessed. Thirty-six studies met these criteria. DATA EXTRACTION: Information was extracted on five topics: physicians' attitudes toward drug industry interactions, frequency with which physicians participate in the interactions, orientation and quality of company-funded clinical trials, content of company-sponsored CME courses and changes in physicians' prescribing behaviour as a result of an interaction. DATA SYNTHESIS: Although most physicians participate only occasionally in company sponsored clinical trials, most see detailers and attend company-sponsored CME courses. However, physicians do not have a very high opinion of the information from detailers or of company-sponsored CME events. Many doctors regard pharmaceutical companies as an important source of funding for clinical trials, but they also have concerns about accepting money from this source. Company funding of clinical trials may affect the quality of the trials and the types of research that physicians undertake. Company-sponsored CME courses may have a commercial bias even if conducted under guidelines designed to ensure the independence of the event. All three types of interactions affect physicians' prescribing behaviour and, in the case of obtaining information from detailers, physicians' prescribing practices are less appropriate as a result of the interaction. CONCLUSIONS: Physicians are affected by their interactions with the pharmaceutical industry. Further research needs to be done in most cases to determine whether such interactions lead to more or less appropriate prescribing practices. The CMA's guidelines on this topic should be evaluated to see whether they are effective in controlling physician-industry interactions. Further measures may be necessary if the guidelines fail to prevent negative effects on prescribing practices. PMID- 8221425 TI - Periodic health examination, 1993 update: 3. Periodontal diseases: classification, diagnosis, risk factors and prevention. Canadian Task Force on the Periodic Health Examination. AB - OBJECTIVE: To review the 1979 Canadian Task Force on the Periodic Health Examination recommendations on the diagnosis and prevention of periodontal disease. OPTIONS: Self-care at home, professional care, treatment with antimicrobial agents and management of patients at high risk. OUTCOMES: Maintaining the gingiva, alveolar bone and periodontal ligament in a healthy state (absence of gingival bleeding and no loss of epithelial attachment). EVIDENCE: A literature search for articles published from 1980 to 1993 was conducted. Selected studies published before 1980 were also reviewed if there were no recent updates. Evidence was evaluated and classified as good, fair or poor according to the criteria of the task force. VALUES: The task force's evidence-based rules for recommendations were used. In addition, whenever feasible, and on the basis of advice from the reviewers and experts, recommendations were modified in the interest of maintaining oral health (e.g., dental flossing in children). BENEFITS, HARMS AND COSTS: The recommendations are not expected to increase the costs of preventing periodontal diseases for the general population. They could benefit dental patients and reduce costs because they are based on the periodontal needs of patients rather than on the current universal application approach. PMID- 8221426 TI - Effects of acetaminophen on adverse effects of influenza vaccination in health care workers. AB - OBJECTIVE: To evaluate the effects of acetaminophen on the incidence of adverse effects to, and the immunogenicity of, whole-virus influenza vaccine in health care workers. DESIGN: Prospective, randomized, double-blind placebo-controlled trial. SETTING: Health Sciences Centre, an acute care teaching hospital in Winnipeg. PARTICIPANTS: Of 474 hospital personnel who agreed to undergo influenza vaccination during the 1990-91 season 262 volunteered to participate in the study. INTERVENTIONS: A dose of 0.5 mL of inactivated trivalent whole-virus influenza vaccine was injected into the deltoid muscle. Volunteers were randomly assigned to ingest two capsules of acetaminophen in a half dose (162.5 mg per capsule) or a full dose (325 mg per capsule) or two identical placebo capsules. Capsules were to be taken at vaccination and at 4, 8 and 12 hours afterward. Subjects were asked to answer questions regarding six symptoms in a diary for the 3 days after vaccination and to record their ingestion of the study medication. MAIN OUTCOME MEASURES: Incidence of local (sore arm) and systemic (headache, fever, muscle ache, nausea and diarrhea) side effects as well as serum titres of hemagglutination inhibition (HAI) antibody to vaccine antigens before vaccination and 2 weeks and 6 months afterward. RESULTS: A total of 87, 87 and 88 subjects received the half dose, full dose and placebo respectively; 96% returned the diaries, 83% ingested all four doses of medication, and 87% volunteered all blood samples. Compared with the placebo group the incidence of sore arm was 25% to 28% lower in the half-dose and full-dose groups respectively at 24 hours after vaccination, and the rate of nausea was 90% lower in the full-dose group. The HAI titres were similar among the groups at the three test times. CONCLUSIONS: The full dose of acetaminophen significantly reduced the incidence of sore arm and nausea without affecting the antibody response. Acetaminophen use may increase the acceptance of influenza vaccine by health care workers in whom concern about side effects is an impediment to vaccination. PMID- 8221427 TI - Case-control study of prenatal ultrasonography exposure in children with delayed speech. AB - OBJECTIVE: To determine whether there is an association between prenatal ultrasound exposure and delayed speech in children. DESIGN: Case-control study. SETTING: Network of community physicians affiliated with the Primary Care Research Unit, University of Calgary. SUBJECTS: Thirty-four practitioners identified 72 children aged 24 to 100 months who had undergone a formal speech language evaluation and were found to have delayed speech of unknown cause by a speech-language pathologist. For each case subject the practitioners found two control subjects matched for sex, date of birth, sibling birth order and associated health problems. MAIN OUTCOME MEASURES: Rates of prenatal ultrasound exposure and delayed speech. RESULTS: The children with delayed speech had a higher rate of ultrasound exposure than the control subjects. The findings suggest that a child with delayed speech is about twice as likely as a child without delayed speech to have been exposed to prenatal ultrasound waves (odds ratio 2.8, 95% confidence limit 1.5 to 5.3; p = 0.001). CONCLUSION: An association between prenatal ultrasonography exposure and delayed speech was found. If there is no obvious clinical indication for diagnostic in-utero ultrasonography, physicians might be wise to caution their patients about the vulnerability of the fetus to noxious agents. PMID- 8221428 TI - Sudden unexpected death in the emergency department: caring for the survivors. AB - OBJECTIVE: To determine whether emergency department staff met the needs of the next of kin and close friends ("survivors") of patients dying in an emergency department and to assess the effectiveness of a program to improve care of survivors. DESIGN: Mail survey before and after program implementation. SETTING: Emergency department of a tertiary care, adult teaching hospital. PARTICIPANTS: Two groups of survivors, identified through a review of emergency department records of deaths during two 6-month periods. In the first group, surveyed in 1987, before program implementation, 26 (53%) of 49 responded; in the second group, surveyed in 1990, after program implementation, 40 (70%) of 57 responded. INTERVENTIONS: A structured, multidisciplinary protocol for notifying next of kin of death and supporting the survivors was implemented. An educational program was provided to all emergency department staff. An information pamphlet was created and provided to survivors. MAIN OUTCOME MEASURES: Questionnaire responses regarding the adequacy and timeliness of information provided, the support and actions by emergency department staff and the survivors' desire to be present during resuscitation efforts. RESULTS: Comparison of responses before and after program implementation showed that adequate information was provided before notification of death in 32% and 83% of cases respectively (p < 0.001), lengthy delays in receiving medical information occurred in 60% and 15% of cases (p < 0.01), adequate medical information concerning the events of death was provided in 53% and 88% (p < 0.05), the presence of emergency department staff was sufficient in 40% and 79% (p < 0.01), survivors spent less than 2 hours in the emergency department in 50% and 81% (p < 0.05), and survivors expressed a desire to be present during resuscitation efforts in 95% and 11% of cases (p < 0.001). CONCLUSION: The grievous experience of learning that a loved one has suddenly and unexpectedly died in the emergency department can be alleviated somewhat by a structured, multidisciplinary approach combined with staff sensitization and education. PMID- 8221429 TI - Dancing around the fountain. PMID- 8221430 TI - Statement on hepatitis B vaccine. National Advisory Committee on Immunization. PMID- 8221431 TI - College bitter as courts quash or reduce MDs' sexual-abuse penalties. PMID- 8221433 TI - In Medellin, the drug wars mean that emergency physicians are always busy. PMID- 8221432 TI - Dalhousie's Pediatric Neurology Group: "We throw rocks at our glass house". PMID- 8221434 TI - Brutal conditions in Somalia, Yugoslavia open eyes of armed forces' medical personnel. PMID- 8221435 TI - Art, play-therapy programs help children whose parents are dying of cancer. PMID- 8221436 TI - Medicine is a family industry in Magrath, Alta. PMID- 8221437 TI - New dictionary provides native-language equivalents of English medical terms. PMID- 8221438 TI - The obstetric care crisis facing Ontario's rural hospitals. PMID- 8221439 TI - Bleak days in Britain. PMID- 8221440 TI - An appeal for simpler medical terminology. PMID- 8221441 TI - Sue Rodriguez and palliative care. PMID- 8221442 TI - Sue Rodriguez and palliative care. PMID- 8221443 TI - CMA policy on alcohol. PMID- 8221444 TI - Pathology: the "unloved" specialty. PMID- 8221445 TI - Pathology: the "unloved" specialty. PMID- 8221446 TI - Evaluating clinical teachers. PMID- 8221447 TI - A rational approach to the cause, prevention and treatment of postdural puncture headache. AB - OBJECTIVE: To review the current research and formulate a rational approach to the cause, prevention and treatment of postdural puncture headache (PDPH). DATA SOURCES: Articles published from January 1980 to April 1992 were obtained through a search of MEDLINE and Index Medicus. Key reference articles published before 1980 were also reviewed. STUDY SELECTION: All pertinent studies were included and critically analysed. DATA SYNTHESIS: PDPH occurs when a slow leak of cerebrospinal fluid leads to contraction of the subarachnoid space and compensatory expansion of the pain-sensitive intracerebral veins. Female sex and an age between 20 and 40 years have been shown to be independent risk factors for PDPH, but pregnancy has not. The rate of PDPH is directly proportional to the diameter of the needle used and also depends on the design of the needle tip. Prophylactic epidural blood patching or saline infusion after dural puncture can decrease the incidence of PDPH, but both are invasive procedures. Intravenous caffeine sodium benzoate therapy effectively relieves PDPH, but the headache may recur. An epidural blood patch is an invasive but effective, permanent treatment for PDPH in most cases; resistant cases may respond to epidural saline infusion. CONCLUSION: The rate of PDPH after lumbar puncture can be minimized through strict attention to technique and the employment of a 25-gauge needle with the bevel parallel to the dural fibres. A reliable diagnosis and stepwise approach to treatment will minimize complications. PMID- 8221448 TI - A measurement process for evaluating clinical teachers in internal medicine. AB - With input from faculty members and residents in the internal medicine residency training program at McMaster University, Hamilton, Ont., the authors developed a form to evaluate attending physicians in clinical teaching units according to 14 domains of performance. Although brief, the form included all main areas of teacher performance, allowed specification of attending physician behaviour and differentiated areas of strengths and weaknesses between and within attending physicians. The authors describe the new evaluation process and the results for 41 attending physicians. PMID- 8221449 TI - Can Haemophilus influenzae type b-tetanus toxoid conjugate vaccine be combined with diphtheria toxoid-pertussis vaccine-tetanus toxoid? AB - OBJECTIVE: To assess the side effects and immune responses after three serial doses of PRP-T vaccine (a Haemophilus influenzae type b [Hib]-tetanus toxoid conjugate vaccine) given concurrently or mixed with adsorbed DPT vaccine (diphtheria toxoid-pertussis vaccine-tetanus toxoid). DESIGN: Multicentre randomized controlled trial. SETTING: Four public health units in western Canada. PARTICIPANTS: Healthy infants 8 to 15 weeks old at entry who were able to receive routine primary vaccinations. Of 444 infants enrolled, 433 (98%) completed the study. INTERVENTIONS: All infants received PRP-T and DPT vaccines at 2, 4 and 6 months of age: half received them mixed in one injection and the others as separate, bilateral injections. MAIN OUTCOME MEASURES: Side-effects 24 and 48 hours after each dose and serologic responses to each vaccine component. RESULTS: Follow-up was obtained after all 1312 vaccinations. Fever was infrequent in the two treatment groups. Local adverse effects of the PRP-T vaccine were infrequent and mild (e.g., redness was noted in 5.9% of cases and the area of redness was more than 2.5 cm in diameter in 0.8%). The incidence rate of local effects of the DPT-containing vaccines was the same in the two groups except for tenderness, which was more frequent in the group given the mixed vaccine (26.6% v. 17.9%, p < 0.001). Serologic data were available for 97% of the subjects. After the three doses 98.1% of the subjects had a PRP antibody level of 0.15 micrograms/mL or more, and 87.9% had a level of 1.0 micrograms/mL or more, both levels compatible with protection against Hib. Responses to PRP-T were comparable between the treatment groups as were responses to the diphtheria and tetanus toxoids. Pertussis agglutinin titres were reduced after administration of one of two PRP-T lots mixed with DPT vaccine, but responses to four other pertussis antigens were not impaired. CONCLUSION: PRP-T vaccine is well tolerated and immunogenic. Combined PRP-T and DPT vaccines performed satisfactorily and may be the preferred method of administration. PMID- 8221450 TI - Outcome of rationing access to open-heart surgery: effect of the wait for elective surgery on patient outcome. AB - OBJECTIVE: To assess the effect of the waiting period before elective open-heart surgery on patient outcomes. DESIGN: Retrospective analysis. SETTING: The Montreal Heart Institute, a referral centre in cardiology and cardiac surgery. PATIENTS: All 568 patients who underwent open-heart surgery on an elective basis or following urgent admission or interhospital transfer between October 1991 and February 1992. MAIN OUTCOME MEASURES: In-hospital death rate, incidence of postoperative complications, length of stay in the intensive care unit (ICU) and total length of hospital stay. RESULTS: A total of 206 patients (151 men and 55 women with an average age of 59.0 [standard error of the mean (SEM) 1] years) underwent elective surgery, and 362 patients (264 men and 98 women with an average age of 62.0 [SEM 1] years) underwent urgent surgery. The mean wait for elective surgery was 2.8 (SEM 0.2) months. There was no significant difference between the two groups in the in-hospital death rate (4% v. 4%), the average length of stay in the ICU (4.4 [SEM 0.2] days v. 5.8 [SEM 1] days) or the average total length of hospital stay (9.0 [SEM 0.4] days v. 9.1 [SEM 1] days). As would be expected, postoperative complications developed in significantly more patients in the urgent group (27%) than the elective group (18%) (p = 0.02). Eight patients were admitted on an urgent basis for surgery owing to worsening symptoms or acute myocardial infarction after a mean wait of 4.6 months. One patient died suddenly at home 1 month after medical investigation while awaiting repeat coronary artery bypass grafting. Among the 206 patients who underwent elective surgery there was no relation between waiting time and adverse clinical outcomes after surgery. CONCLUSIONS: The results suggest that the wait before elective open-heart surgery had no effect on patient outcome after surgery in our institution. A policy of a short waiting period before elective open-heart surgery for patients whose condition is stable is safe and acceptable only if rapid access to medical and surgical treatment is available should it become necessary. PMID- 8221451 TI - Angioplasty redeemed? PMID- 8221452 TI - Statement on pertussis immunization. National Advisory Committee on Immunization. AB - The following recommendations on pertussis vaccine from the National Advisory Committee on Immunization will appear in the fourth edition of the Canadian Immunization Guide, scheduled for publication in autumn 1993. These revised recommendations represent a major change in vaccine usage and take account of the current understanding of adverse events associated with use of pertussis vaccine. Most adverse events are no longer contraindications to further immunization; only anaphylactic reaction to a previous dose remains an absolute contraindication to pertussis immunization. Other adverse reactions for which there are no sequelae or that have not been proven to be caused by the vaccine are no longer considered valid reasons for withholding immunization. It is hoped that simplification of the contraindications will improve vaccine utilization and achieve higher levels of age-appropriate immunization. PMID- 8221453 TI - Medicine and misogyny. PMID- 8221454 TI - The remarkable healing power of a $20 bill. PMID- 8221455 TI - Medication use and the elderly. Canadian Medical Association. PMID- 8221456 TI - Canada's first private MRI clinic: does it signal a shift to two-tiered medicine? PMID- 8221457 TI - Women may be "their own worst enemies" as they attempt to climb medical ladder. PMID- 8221459 TI - Retiring president warns of "chaos" facing Canada's health care system. PMID- 8221458 TI - Study of lung cancer treatment costs may point to savings on a broader scale. PMID- 8221461 TI - Hepatitis B immunization project in BC shows positive results. PMID- 8221460 TI - Nurses may become key players in providing primary care to Canadians, CPHA meeting told. PMID- 8221462 TI - Canadian microbiologist spreads message about danger of rotaviral infections. PMID- 8221463 TI - HIV testing and counselling. PMID- 8221464 TI - More on hospital foundations. PMID- 8221465 TI - Breast implants. PMID- 8221466 TI - Drug patent law reform and shortages. PMID- 8221467 TI - Future for new MDs. PMID- 8221468 TI - Assisted reproductive technology. PMID- 8221469 TI - Standards of aerosol therapy. PMID- 8221470 TI - Improving the lot of emergency physicians. PMID- 8221471 TI - Drug advertising. PMID- 8221472 TI - Compensating occupational diseases. PMID- 8221473 TI - Primary prevention of neural tube defects: notice from the HPB. PMID- 8221474 TI - Isoniazid and pyridoxine. PMID- 8221475 TI - Low-birth-weight infants and breast-feeding. PMID- 8221476 TI - Unrecognized adverse drug reactions. PMID- 8221477 TI - Physical abuse in pregnancy: what can we do about it? PMID- 8221478 TI - Recommendations on the use of folic acid supplementation to prevent the recurrence of neural tube defects. Clinical Teratology Committee, Canadian College of Medical Geneticists. AB - OBJECTIVE: To prevent the recurrence of neural tube defects (NTDs) in families at increased risk of having offspring with NTDs with the use of periconceptional folic acid supplementation. OPTIONS: Genetic counselling and prenatal diagnosis of NTDs. OUTCOMES: NTDs cause stillbirth, neonatal death and severe disabilities. The cost for medical care and rehabilitation in the first 10 years of life of a child with spina bifida cystica was estimated to be $42,507 in 1987. EVIDENCE: The authors reviewed the medical literature, communicated with investigators from key studies, reviewed policy recommendations from other organizations and drew on their own expertise. A recent multicentre randomized controlled trial showed that among women at high risk of having a child with an NTD those who received 4 mg/d of folic acid had 72% fewer cases of NTD-affected offspring than nonsupplemented women. Two previous intervention studies also demonstrated that folic acid supplementation was effective in reducing the rate of NTD recurrence. Several retrospective studies support this conclusion. VALUES: Recommendations are the consensus of the Clinical Teratology Committee of the Canadian College of Medical Geneticists (CCMG) and have been approved by the CCMG Board. The committee believes that primary prevention of NTDs is preferable to treatment or to prenatal detection and abortion. BENEFITS, HARMS AND COSTS: Folic acid supplementation should result in fewer NTDs among infants in Canada and ancillary savings in medical costs. The recommended dosage of folic acid is not known to be associated with adverse effects. Higher dosages of folic acid may make vitamin B12 deficiency difficult to diagnose and may alter seizure frequency in patients with epilepsy due to drug interactions with anticonvulsants. RECOMMENDATIONS: A minimum dosage of folic acid of 0.8 mg/d, not to exceed 5.0 mg/d, is recommended along with a well-balanced, nutritious diet for all women who are at increased risk of having offspring with NTDs and who are planning a pregnancy or may become pregnant. Supplementation should begin before conception and continue for at least 10 to 12 weeks of pregnancy. VALIDATION: These guidelines are similar to those of the Society of Obstetricians and Gynaecologists of Canada, the US Centers for Disease Control and Prevention and the Department of Health in Britain. SPONSORS: These guidelines were developed by the CCMG Clinical Teratology Committee and endorsed by the Board of the CCMG. No funding for the development of these guidelines was obtained from any other sources. PMID- 8221480 TI - Physical abuse in pregnancy. AB - OBJECTIVES: To determine the prevalence of physical abuse during late pregnancy and to investigate how abused and nonabused pregnant women differ in demographic characteristics, health habits, psychologic distress and attitudes about fetal health. DESIGN: Survey of women attending for prenatal health care or admitted to hospital for delivery. The information was obtained on one occasion from self report questionnaires, completed with the option of anonymity. SETTINGS: Community-based prenatal clinic, private obstetricians' offices in a large city, private family physicians' offices in a large city, family physicians' offices in a small town, and a university teaching hospital. PATIENTS: English-speaking women at 20 weeks' or more gestation attending or admitted consecutively. INTERVENTIONS: Three self-report questionnaires: the General Health Questionnaire (GHQ), the Fetal Health Locus of Control (FHLC) and the study questionnaire. RESULTS: Thirteen women (2.4%) refused to participate in the survey. Of the 548 women who completed the questionnaires 36 (6.6%) reported physical abuse during the current pregnancy and 60 (10.9%) before it. There were no significant differences in rates of abuse between settings. Of the women abused during the pregnancy 23 (63.9%) reported increased abuse during the pregnancy, and 28 (77.8%) remained with the abuser. Twenty-four pregnant women (66.7%) received medical treatment for abuse, but only 1 (2.8%) told her prenatal care provider of the abuse. Factor analysis revealed three factors associated with physical abuse in pregnancy: "social instability" (comprising low age, unmarried status, lower level of education, unemployment and unplanned pregnancy), "unhealthy lifestyle" (comprising poor diet, alcohol use, illicit drug use and emotional problems) and "physical health problems" (comprising health problems and prescription drug use). The GHQ scores showed that the abused women were significantly more emotionally distressed than the nonabused women (p < 0.001). The FHLC scores showed that the abused women believed they had little "internal control" over the health of their fetuses and that "chance" played the most important role in the outcome of their pregnancy (p < 0.001). CONCLUSIONS: Abused pregnant patients are a frequently undetected high-risk group. Prenatal care should include a routine screening question about domestic violence, and identified patients should be appropriately counselled and referred. PMID- 8221479 TI - Why don't we use vitamin E in dermatology? AB - OBJECTIVE: To review the possible uses of topical and systemic tocopherols as therapy for skin conditions in light of the widespread use of vitamin E by patients. DATA SOURCES: Index Medicus was searched for articles published from 1922 (when vitamin E was discovered) to 1966 (the beginning of MEDLINE). MEDLINE was searched for articles in English and French on vitamin E or tocopherol in relation to dermatology. Additional original articles were identified from the reference lists of the review articles. STUDY SELECTION: Only well-designed controlled studies were accepted; anecdotes and open studies are cited for completeness and as direction for future research. DATA SYNTHESIS: There was some weak or conflicting evidence that vitamin E is of value in yellow nail syndrome, vibration disease, epidermolysis bullosa, cancer prevention, claudication, cutaneous ulcers, and collagen synthesis and wound healing. It was of no use in atopic dermatitis, dermatitis herpetiformis, psoriasis, subcorneal pustular dermatosis, porphyrias and skin damage induced by ultraviolet light. CONCLUSIONS: After 44 years of research there is still scant proof of vitamin E's effectiveness in treating certain dermatologic conditions. Further research in well-designed controlled trials is needed to clarify vitamin E's role. PMID- 8221481 TI - Chlamydia trachomatis infection in rural Nova Scotia. AB - OBJECTIVE: To examine the demographic characteristics of patients who underwent testing for Chlamydia trachomatis and to determine the clinical and behavioural characteristics and the types of treatment for those who had positive test results. DESIGN: Case series. SETTING: Rural county in Nova Scotia. PATIENTS: All residents of the county for whom testing for C. trachomatis was ordered at the regional hospital from Sept. 1, 1990, to Mar. 31, 1991. MAIN OUTCOME MEASURES: Rates of testing and of positive test results by age and sex. Comparison of patient and physician characteristics in relation to testing rates. RESULTS: Of the 1116 patients tested 58 (5.2%) had positive test results. Females accounted for 82.8% of those with positive results whose sex could be determined. Among the females the mean age of those with a positive result was 22.3 years, as compared with 27.5 years for those with a negative result (p < 0.0001). Females 15 to 19 years of age were less likely to have a test performed than women 20 to 29 years and were more likely to have a positive test result than the women in the older groups. Almost 9% of the testing among the females was in those over 39 years of age, although no infection was seen in this age group. The number of tests ordered per general or family practitioner varied from 1 to 154; the physicians' sex, practice location and length of time in practice did not predict the rates of positive test results. Treatment was most often in keeping with that recommended by national guidelines. Four (8.5%) of the 47 patients with positive results who were interviewed were not aware of their diagnosis, either because they had not returned for follow-up or had not being notified by the physician's office. CONCLUSIONS: The frequency of testing for C. trachomatis infection may be less than is desirable among young patients, who, if tested, are more likely than older patients to have positive results. More understanding of the diagnostic approach taken by physicians is needed. PMID- 8221482 TI - Adjuvant radiotherapy for early breast cancer: patterns of practice in Ontario. AB - OBJECTIVE: To determine the number of different radiation schedules used in Ontario to treat women with node-negative breast cancer after lumpectomy and axillary dissection. DESIGN: Retrospective survey. SETTING: Princess Margaret Hospital, Toronto, and regional centres of the Ontario Cancer Treatment and Research Foundation (in Hamilton, London, Ottawa, Windsor and Thunder Bay). PATIENTS: A total of 551 of 1624 consecutive patients with node-negative breast cancer having undergone lumpectomy and axillary dissection who were eligible but did not participate in the Ontario Clinical Oncology Group randomized clinical trial and who received adjuvant breast irradiation between April 1984 and February 1989. OUTCOME MEASURES: Schedules of radiotherapy received. RESULTS: Forty-eight different radiotherapy schedules were identified. Total doses ranged from 4000 to 6600 cGy and the number of fractions from 15 to 30. Several different schedules were preferred: 322 patients (58.5%) received 4000 cGy in 15 or 16 fractions to the whole breast over 3 weeks plus a local boost of 1250 cGy to the primary site in 5 fractions over 1 week; 66 patients (12.0%) received 4000 cGy in 15 or 16 fractions over 3 weeks to the whole breast plus a local boost of 1000 cGy to the primary site in 4 or 5 fractions over 1 week; and 63 patients (11.5%) received 5000 cGy in 25 fractions to the whole breast in 5 weeks, without a boost. CONCLUSIONS: The practice of adjuvant radiotherapy for early breast cancer in Ontario varies. The optimal radiation regimen for patients after lumpectomy should be determined through randomized clinical trials. PMID- 8221483 TI - Fetal outcome after maternal radiation treatment of supradiaphragmatic Hodgkin's disease. PMID- 8221484 TI - Finding the genes of neurologic and psychiatric illnesses: variations on a theme. PMID- 8221485 TI - Questions about sleep should be routine part of patient visits, physician says. PMID- 8221486 TI - "You couldn't beat the medical corps". PMID- 8221487 TI - Canadians ignoring knowledge that they can improve their health: survey. PMID- 8221488 TI - CMA continues development of euthanasia policy after Supreme Court's Rodriguez ruling. PMID- 8221489 TI - Pamphlet on HIV and athletes dispels myths about possibility of infection through sports. PMID- 8221490 TI - Social contracts and health care. PMID- 8221491 TI - Dr. Alexander Augusta: breaking down the barriers for black doctors. PMID- 8221492 TI - Suicide: a public health concern? PMID- 8221493 TI - Suicides on Ontario farms. AB - PURPOSE: To describe the epidemiology of suicides on Ontario farms from 1980 1989, and the ecological relationship between economic strain and farm suicide risk. METHOD: Farm suicides were identified using a computer-assisted search of vital records. Distributions of farm suicides were examined by several demographic variables. Linear and Poisson regression were used to describe potential associations between ten economic indicators and the incidence of farm suicide. RESULTS: 126 farm suicides were identified. After adjustment for age and under-reporting, the farm suicide rate was estimated at 7.2 per 100,000 per year. The highest suicide rate was observed among elderly farm owner-operators. Regression analyses failed to find any associations between the economic indicators and farm suicide rates. CONCLUSION: The results do not support the notion that farmers in Ontario generally have a high rate of suicide, the ecological analysis provided no evidence for increased farm suicide risk with increasing economic strain. PMID- 8221494 TI - Elderly suicide in British Columbia: an exploration of regional variation and related factors. AB - In 1986, the Canadian Task Force on Suicide reported coast-to-coast variations in suicide rates across Canada. This review of 1,040 reported suicides in British Columbia between 1985 and 1988 shows variations in suicide rates that are related to population size. Elderly suicides are disproportionate in some, but not all, regions. Urban population centres have lower suicide rates than low population areas. Explorations of the factors which explain these variations may provide understanding of suicide and its prevention. PMID- 8221495 TI - Community development and partnerships. PMID- 8221496 TI - Population-based health promotion: a new agenda for public health nurses. AB - The role of Public Health Nurses (PHNs) involves participation in the development and implementation of population-based health promotion activities. To support encourage and facilitate such participation, the Nursing Division of the Hamilton Wentworth Department of Public Health Services initiated a "New Agenda" that would help to realize these PHN roles. The New Agenda involved a commitment by the Nursing Division to support population-based health promotion activities by PHNs through changes in Nursing Division policies and practices, and the advancement of a consensus among PHNs regarding the practice of population-based health promotion. A series of four workshops were designed and introduced to ensure that structural barriers to developing PHN roles were eliminated. PHNs stated they achieved role clarification and role acceptance through the New Agenda interventions. PMID- 8221497 TI - The incidence of wife abuse and mental health status in abused wives in Edmonton, Alberta. AB - To determine the incidence and type of wife abuse in relation to mental health status, a survey was undertaken in a community-based sample of wives. The incidence rate of physical abuse was 10.6%, psychological abuse 13.1%. Nearly all physically abused wives were psychologically abused. Wives at highest risk of abuse were those: 1) separated from their husbands in the previous year; 2) aged 18-44 years; 3) in relatively new relationships; 4) whose husbands were unemployed, in school or working part-time. Physically and psychologically abused wives had more somatic complaints, higher levels of anxiety and insomnia, greater social dysfunction and more symptoms of depression than non-abused wives. Alcohol dependency was associated with abuse; 16.3% of physically abused and 11.3% of psychologically abused wives were alcohol-dependent, compared to 2.4% of non abused wives. PMID- 8221498 TI - Promoting social health in northern Saskatchewan. AB - In response to concern about social health problems in Northern Saskatchewan, a Working Group on Social Health was established in 1989 in the Research and Development Committee of Northern Medical Services. The Group formulated a concept of mental health in social terms; found and interpreted indicators of the extent of social health problems; identified major determinants of social health problems, barriers to effective coping and problems in providing adequate support and services; and identified strategies and program models that could be more effective in promoting social health in this region. Indicators of problems and underlying determinants are discussed, along with strategies for change. These strategies are based on a community development model, and incorporate innovation and reaffirmation of values and ways that have traditionally given people strength. PMID- 8221499 TI - Environment and well-being before and following smoking ban in office buildings. AB - The purpose of this study was to compare the well-being, salivary cotinine levels and environmental characteristics of office building workers and their workplace before and after the introduction of a smoking ban. The results showed that one year following the ban, there was an ambiguous change in symptom reporting, a trend towards reduced salivary cotinine levels, and an improvement in measured but not in perceived environmental quality. PMID- 8221500 TI - The validity of a province-wide student drug use survey: lessons in design. AB - This study assesses the validity and reliability of a province-wide survey on drug use by high school students in Nova Scotia in 1991. The Nova Scotia instrument was derived from the prototype provided in the Canadian guidelines for self-reported adolescent drug use surveys. To our knowledge, this is the first study on the validity and reliability of the Canadian instrument, as applied in a province, since 1978. Two new methods are proposed as validity and reliability checks: a statistical method to assess possible exaggeration by the group claiming to have used a fictitious drug, and a method to review the entire instrument based on responding error distributions. Overall, this study provides strong evidence of the validity and reliability of the items specifically about drug use. The major source of error uncovered using the responding error distributions, was related to instrument design and occurred in items considered to be explanatory variables for drug use. PMID- 8221501 TI - Meeting a person with AIDS in the classroom: an evaluation. AB - Meeting a person with AIDS in the classroom was evaluated to determine if it had an impact on students' perceived susceptibility to HIV infection and attitudes toward persons with AIDS. The meeting was incorporated into the Grade 9 AIDS education program of a school district in Nova Scotia. Four schools participated in this study. Two schools were randomly assigned to the treatment group, which met the person with AIDS, and the remaining two schools formed the comparison group. Measures of the two attitudinal variables were collected using a self report questionnaire that was administered both prior to and two weeks after the educational intervention. Meeting a person with AIDS in the classroom had no measurable impact on students perceived susceptibility to HIV infection nor on their attitudes toward persons with AIDS. Suggestions for using the educational intervention more effectively and for further research are made. PMID- 8221502 TI - [Adolescents and AIDS: knowledge, attitudes, behaviors, perceptions and consequences for prevention]. AB - The vital issues at stake in the credibility and effectiveness of AIDS prevention programs for young people have led to the emergence of a trend in research focused on the study of adolescents' understanding of AIDS, their attitudes and behaviour towards it, and their beliefs about it. This research can be classified under the three main ways of approaching the problem: 1) results of inquiries about knowledge, attitudes and behaviour; 2) research centred on beliefs and their consequences on a social and individual level; 3) reports and analysis of prevention programs. The results obtained by the different authors bring to light certain characteristics among the adolescent population, notably, their understanding of the disease, their attitudes towards people who are HIV positive, their sexual behaviour, their beliefs about AIDS, and the general direction of the prevention programs which are best adapted to this particular population. PMID- 8221503 TI - Changes in university students' AIDS-related knowledge, attitudes and behaviours, 1988 and 1992. AB - We evaluated changes in AIDS-related knowledge, attitudes and behaviours among students in residence at the University of British Columbia based on identical surveys conducted in 1988 and 1992. 493 randomly selected students in each year were sent a self-administered questionnaire on AIDS-related knowledge, fear of AIDS, homophobia, sexual behaviours, and intravenous drug use. AIDS-related knowledge was very high and increasing (p < 0.01) among respondents. Fear of AIDS and homophobia were low in both years. Intravenous drug use was reported by only one student in each year. Sexual activity was reported by 62% of students overall, with 25% of these reporting multiple sexual partners in the previous six months. 23% of sexually active students in 1988 and 40% in 1992 reported usually or always using condoms during intercourse. In spite of high levels of AIDS related knowledge, a large, although decreasing, proportion of students have multiple partners without barrier protection. Nevertheless, the consistent presentation of the same public health message over several years is associated with substantial improvement in condom use. PMID- 8221504 TI - Hepatitis B prenatal screening survey, Nova Scotia, 1990-1991. AB - We conducted a study in which 5,754 pregnant women who delivered at the Grace Maternity Hospital in Halifax were screened for HBsAg. There were five who were found to be seropositive for the first time (a screening yield for seropositivity of 8.7/10,000). Overall six were seropositive for a prevalence rate of 10.4/10,000. These rates are above the 6.0/10,000 level at which routine prenatal screening is considered to be cost-effective. Screening based upon risk factors would have identified only two of the five women who were found for the first time to be HBsAg seropositive. Based upon the results of this study, we recommend that routine screening for HBsAg be performed on all prenatal women in Nova Scotia. PMID- 8221505 TI - The incidence of cervical cancer among Chinese and Caucasians in British Columbia. PMID- 8221506 TI - Seafood allergy. Introduction. PMID- 8221507 TI - Phylogeny of commercially important seafood and description of the seafood industry. PMID- 8221508 TI - Allergic reactions to fish. AB - A wide variety of fish are known to induce allergic reactions following ingestion or inhalation of vapors by sensitized individuals. Although the exact prevalence of fish sensitivity is not known, fish are among the most important food allergens; and as consumption of fish increases, rates of sensitization are expected to increase. Diagnosis of fish allergy is aided by clinical history, skin prick testing, and in vitro assays; however, double-blind placebo-controlled food challenges are the most reliable method to confirm fish allergy and to identify putative species. It appears from RAST inhibition and SDS-PAGE/Western blot studies that the current policy of recommending that fish-sensitive individuals avoid all species of fish should be reevaluated. The major allergen in codfish (Gad cI) is one of the most extensively studied of all allergens. It is a calcium-chelating protein, with a mol wt of 12,328 kDa and an isoelectric point of 4.75. As an allergen, Gad cI is very stable and its allergenic activity appears to be dependent on amino acid sequence. Crossreactivity among some fish species may be the result of common structures within related proteins. PMID- 8221509 TI - Hypersensitivity reactions to crustacea and mollusks. PMID- 8221510 TI - Occupational reactions in the seafood industry. PMID- 8221511 TI - Seafood toxins. PMID- 8221513 TI - Lyme borreliosis. PMID- 8221512 TI - Chemical additives in seafood products. PMID- 8221514 TI - How the discovery of Borrelia burgdorferi came about. PMID- 8221515 TI - Epizootiology of Lyme disease-causing borreliae. PMID- 8221516 TI - Erythema migrans. PMID- 8221517 TI - Role of Borrelia burgdorferi in lymphocytomas and sclerotic skin lesions. PMID- 8221518 TI - Acrodermatitis chronica atrophicans. PMID- 8221519 TI - Histopathologic patterns of erythema migrans and borrelial lymphocytoma. PMID- 8221520 TI - Histopathologic patterns of acrodermatitis chronica atrophicans. PMID- 8221521 TI - Neurologic manifestations in Lyme borreliosis. PMID- 8221522 TI - Rheumatic manifestations in Lyme borreliosis. PMID- 8221523 TI - Laboratory diagnostic methods in Lyme borreliosis. PMID- 8221524 TI - Immunopathogenesis of Lyme borreliosis. PMID- 8221526 TI - CNSs stretch an inadequate health care system. PMID- 8221525 TI - Therapeutic aspects of Lyme borreliosis. PMID- 8221527 TI - Incorporating imagery in clinical practice. PMID- 8221528 TI - Imagery: a strategic intervention to empower clients. Part II--A practical guide. PMID- 8221530 TI - Legislative and regulatory update: reimbursement and health care reform legislation introduced. PMID- 8221529 TI - The counterpart relationship: a model for clinical nurse specialists in international nursing practice. AB - The proposed counterpart relationship model can be an effective tool in blending role functions to provide culturally sensitive nursing care in international settings. The authors explore their experience in developing a counterpart relationship where the CNS enters into a mutually collaborative relationship with a host counterpart from a cultural base different from the CNS's. The goal is to improve the level of health care provided to a specific cultural group and to transition the responsibilities of the CNS to the host counterpart. PMID- 8221531 TI - Measuring the outcomes of our practice. PMID- 8221532 TI - An empirical study of a model of self-care in chronic illness. AB - A descriptive correlational study was conducted to test the efficacy of the Model of Self-Care in Chronic Illness. The model, an adaption and extension of the Health Belief Model, was developed to describe and relate the variables that influence effective self-care among ambulatory chronically ill adults. Participants (n = 181) were randomly selected from the waiting lists of an ambulatory care clinic of a major military medical center. The research examined general self-care, i.e., behaviors to promote health and prevent illness, and therapeutic self-care, i.e., education, diet, and other behaviors prescribed for a specific illness. Self-concept was correlated significantly with general as well as medication and dietary self-care. Psychological status and general health motivations influence general self-care. Stepwise multiple regression analyses demonstrated that costs of medication, patients' psychological status and perceptions of the seriousness of their illnesses, and vulnerability to complications were significantly related to medication self-care, while costs and perceptions of seriousness and vulnerability were significantly related to dietary self-care. PMID- 8221533 TI - Latex allergy: collaborative nursing research using a consortium model. AB - Nursing research, a recognized component of the CNS role, is difficult to develop in practice. Collaborative research is frequently recommended as a way to promote research activity by advanced practice nurses. Although most collaborative nursing research involves cooperation between academic and clinical nurses or within nursing organizations that exist for the purpose of conducting research, a regional nursing consortium can successfully carry out research activities. An investigation into the incidence and nature of latex allergy in children with myelodysplasia grew out of the education, mutual support, and advocacy activities of a regional clinical consortium. This article shares the research process, the obstacles encountered, and the strategies employed to identify, investigate, and use new nursing knowledge. PMID- 8221534 TI - The ultimate goal. PMID- 8221535 TI - The role of the psychosocial oncology CNS in a rural cancer outreach program. AB - The high cancer mortality rates in rural areas of Virginia are discussed, and cancer risk factors particular to the rural population are delineated. Rural health beliefs and their relationship to delay in seeking cancer care in rural areas are examined. A rural cancer outreach program and the role of the oncology clinical nurse specialist (CNS) in the program are described in terms of Psychosocial Oncology CNS subroles, i.e., clinician, consultant-liaison, networker/linker, coach, educator, change agent, researcher, and supervisor/supervisee. Effectiveness of the role in the program is analyzed. An innovative urban-rural nurse oncology program is delineated, and recommendations for research in rural nursing cancer care and actions for fostering CNS role development are suggested. PMID- 8221536 TI - Can the CNS survive with managed competition? PMID- 8221537 TI - Capturing the multiple contributions of the CNS role: a criterion-based evaluation tool. AB - The CNSs at a community hospital found that tools used to evaluate their performance did not reflect the wide variety of activities native to the CNS role within the institution. A six-member working group came to consensus on practice behaviors, and ordered them into a hierarchy of performance levels. The resulting role description and evaluation tool provides flexibility in application to unit based, centralized, and specialty-based positions. The benefits of the new role description and evaluation tool to the organization and individual are multiple and serve to highlight the unique talents the role can bring to nursing. PMID- 8221538 TI - Development of a surgical/trauma intermediate care unit. AB - During these times of escalating health care costs and increased demand for critical care beds, hospitals are often unable to provide the intensive level of nursing care that is required. One large urban trauma center found that 36% of the surgical intensive care unit patients required a level of nursing care intermediate between the intensive care unit and the general ward. A multidisciplinary team met to plan and develop a surgical/trauma intermediate care unit to provide this level of care. Goals, objectives, specific admission, discharge criteria, and administrative responsibility were determined. An educational program was developed based on a needs assessment of the staff. During the implementation phase and throughout the unit's operation, advanced practice nurses performed an integral role as resource personnel. The purpose of this paper is to describe the multidisciplinary process used in the development of a new unit and to highlight the advanced practice role in this activity. PMID- 8221539 TI - Potential problems in patient support groups: a reason for consultation. PMID- 8221540 TI - When the CNS needs help establishing a women's support group: getting the most from the consultation process. AB - Support groups are an effective intervention strategy to address the unique psychosocial needs of women in transition. The task of establishing a support group can be overwhelming for the CNS who possesses expertise in women's health but not in establishing a support group. Securing the services of an external nurse consultant with experience in group process and project management skills is an invaluable resource to the CNS in charge of developing a formal proposal, setting goals and planning actions, and/or implementing and evaluating interventions. The CNS's role as a consumer of the formal consultation process is addressed. PMID- 8221541 TI - Epidemiology of soft-tissue and bone tumors of the foot. AB - This article reviews and discusses the epidemiology of soft-tissue and bone tumors of the foot. Specific reference is made to their frequency and location of occurrence as well as the age and gender of patients. Although the relative frequency of soft-tissue and bone tumors of the foot is low, their occurrence is still frequent enough to warrant serious consideration. PMID- 8221542 TI - Evaluation of a patient for pedal neoplasia. Basic principles and procedures. AB - The process of determining a diagnosis for a patient with a tumor mass is not easy. It demands astuteness on the part of the attending doctor as well as cooperative consultation and teamwork with radiologists, pathologists, and other specialists. It also demands a basic understanding of the characteristics of benign and malignant tumors in the foot, whether primary or metastatic. This is best accomplished by continued reading of a variety of journals and current texts as well as by attendance at clinical conferences. Concern for the patient must motivate any practitioner to know, understand, and undertake definitive diagnostic procedures to determine the nature and extent of a neoplasm. The practitioner must acquire and maintain an appropriately high index of suspicion and an awareness that the foot can be the site of a primary or metastatic tumor. PMID- 8221543 TI - Imaging methods for assessment of pedal soft-tissue neoplasms. AB - The advent of cross-sectional, tissue-specific, noninvasive multiplanar imaging methods has enabled the radiologist to probe the etiopathogenesis of soft-tissue lesions of the foot to greater advantage than in the past. High-resolution, high contrast images are now available so that a wide variety of conditions may be explored, resulting in accurate compartmental localization and assessment of both adjacent tissue as well as vascular invasion and distant metastasis. PMID- 8221544 TI - Radiographic evaluation of pedal osseous tumors. AB - Osseous tumors of the foot are uncommon lesions. The approach to making the correct diagnosis should be based upon a knowledge of the types of tumor that may be encountered and a systematic analysis of the characteristics of the lesion. Proper use of imaging techniques will help to narrow the differential diagnosis. Consultation among colleagues is very important for diagnosing a specific lesion, for broadening one's knowledge base, and for improving patient care. Topics discussed in this article include fluoroscopy, magnification views, and characteristics of lesions and patterns of destruction. PMID- 8221545 TI - Advanced diagnostic imaging techniques for pedal osseous neoplasms. AB - It is a widely held view that the initial evaluation of foot and ankle osseous tumors and tumor-like lesions begins with the conventional radiograph. It is equally recognized that musculoskeletal radiology has undergone a veritable revolution in the last few decades, as it continues to experience refinements that allow increased sensitivity of anatomic localization, specificity of pathologic tissue characterization, and hence greater insight into the subsequent therapeutic maneuvers that may significantly affect case management and survival outcomes of more aggressive disease processes. This article discusses the basics of various imaging modalities and applies these principles to a number of examples of osseous tumors with the hope that podiatrists, orthopedists, radiologists, and others might develop an approach to selecting the most appropriate diagnostic study, as well as an increased awareness of the appearance of some of the entities that these specialists might encounter in routine practice. PMID- 8221546 TI - Differential diagnosis of pedal osseous neoplasms. AB - Topics discussed in this article include osteogenic sarcoma, osteoid osteoma, osteoblastoma, chondrosarcoma, chondromyxoid fibroma, chondroblastoma, chondroma, Ollier's disease, Maffucci's syndrome, osteochondroma, hereditary multiple exostoses, unicameral bone cyst, fibrous dysplasia, Albright's syndrome, nonossifying fibroma, giant cell tumor, Ewing's sarcoma, and metastasis. Numerous radiographs, CT scans, MR images, arteriograms, and photomicrographs supplement the text. PMID- 8221547 TI - Management of soft-tissue and bone tumors of the foot. AB - Neoplasms of the foot are relatively rare problems. Because of their rarity, many physicians are not familiar with their diagnosis or treatment. In addition, there is a diversity of opinions about the histogenesis, nomenclature, and treatment of these tumors. A systematic approach to evaluation, diagnosis, and treatment is necessary to ensure prompt, appropriate management of these problems. Topics discussed in this article included the initial evaluation, staging, biopsy, surgical resection, and postoperative care. PMID- 8221548 TI - Pigmented villonodular synovitis of multiple tarsal bones. A case report. AB - PVNS because of its indolent symptomatology can present with confounding clinical, radiographic, and pathologic findings. The case study detailed in this article demonstrates the need in most cases to excise clean margins with the involved tissue for better pathologic diagnosis. Currently, the patient described in this article has been followed for over 2 years with no recurrence and has a weight-bearing limb. PMID- 8221549 TI - Hemangiopericytoma of the foot. A case report. AB - Soft-tissue sarcoma is a rare diagnosis in general and more so in the foot. The author has seen two in the last 9 years, both of which were fatal. As podiatry becomes more and more an accepted primary care specialty, our index of suspicion needs to increase and our diagnostic procedures need to be aggressive. The incidence of primary and secondary cancerous lesions in the foot and ankle are increasing. PMID- 8221550 TI - Rhabdomyosarcoma. A podiatric challenge. AB - The clinical case presented in this article demonstrates the necessity of prompt treatment of suspicious mass lesions and highlights the importance of always sending tissue specimens for proper evaluation. In this case, the prognosis was directly related to extent of the disease. Delaying treatment or improper treatment could easily mean the difference between life and death. When a highly aggressive rhabdomyosarcoma is encountered, a planned formal excision with adequate margins is mandatory to achieve optimal results. Multidisciplinary care, including oncology, radiology, pathology, and podiatry services, is preferred. A current 4-year follow-up of the patient discussed in this article has demonstrated no recurrence. PMID- 8221551 TI - Localized prostate carcinoma. Surgical management. AB - Total prostatectomy in the properly selected patient will provide disease-free survival rates comparable to the expected survival in similarly aged men for up to 30 years of observation (Figure 4). Patients who undergo total prostatectomy accept a very small risk of long-term permanent complications or mortality, and effective treatment is available for most complications. The morbidity and costs associated with hormone refractory metastatic prostate cancer are significant, with bone pain and anemia from bone marrow invasion, bladder dysfunction (retention, incontinence, and hematuria), urinary tract infection, anorexia, and uremia from obstructed ureters being common sequelae in the months before death. In the properly selected patient, minimal risk is incurred from total prostatectomy, the potential complications are well defined and manageable, and long-term disease-free survival is seen in most patients. PMID- 8221552 TI - Cervical, mediastinal, and abdominal lymph node dissection (three-field dissection) for superficial carcinoma of the thoracic esophagus. AB - BACKGROUND: The recent increase in the number of esophageal carcinomas detected at an early stage has prompted debate about the most suitable treatment for them. METHODS: Forty-three patients with superficial esophageal carcinoma (SEC) underwent esophagectomy with cervical, mediastinal, and abdominal lymph node dissection (three-field dissection). On the basis of the pathologic data and results of surgery, the adequacy of treatment was discussed. RESULTS: Three patients with pTis tumor had no positive lymph nodes. Twenty patients (46.5%) had positive nodes in the resected specimen. Nineteen lesions (57.6% of submucosal cancers) with lymph node metastasis had invasion to the submucosa. Five patients with submucosal cancer (15.2% of submucosal cancers) had positive nodes in the neck. The lymph nodes along the right recurrent nerve and the right paracardiac nodes were the most frequent site of metastasis (16.3%), whereas the right paratracheal, infracarinal, infra-aortic arch, common hepatic, and celiac nodes had no metastasis. The operative mortality rate was 2.3%. Recurrent disease occurred in three patients with submucosal cancer. The 5-year survival rate after surgery was 73.2% for all 43 patients and 68.6% for patients with positive nodes. CONCLUSIONS: Esophagectomy with three-field lymph node dissection is recommended for patients with lesions diagnosed as submucosal cancer. Endoscopic mucosectomy is applicable as a first-choice treatment for patients with esophageal carcinoma at Tis. PMID- 8221553 TI - Thymidine uptake in vitro as a prognostic indicator for primary gastric cancer. AB - BACKGROUND: Prognostic significance of in vitro thymidine uptake by cancer cells remains unclear in patients with gastric cancer. METHODS: In 173 patients with operable gastric cancer, the relations between thymidine uptake by gastric cancer cells in semi-solid media and their clinicopathologic features as well as their survival lengths were studied. RESULTS: There were significant correlations between in vitro thymidine uptake and such clinicopathologic features as lymph node metastasis (P = 0.00002), lymphatic invasion (P = 0.003), vessel invasion (P = 0.006), peritoneal metastasis (P = 0.010), depth of invasion (P = 0.011), and hepatic metastasis (P = 0.032). Ninety-five of 173 cancers (54.9%) that incorporated 1000 or more cpm in a single well were designated as being a high uptake group. Other gastric cancers (78 of 173; 45.1%) were designated as being a low uptake group. The overall survival rate of the patients was demonstrated to be significantly longer in the group with a low thymidine uptake than with a high uptake (P < 0.00001). The multivariate analysis showed that thymidine is one of the two variables that are the most highly correlated with the probability of patient death (P = 0.00044). CONCLUSIONS: These results indicated that in vitro thymidine uptake is an independent prognostic parameter for gastric cancer and may be useful for selecting patients who would benefit from more intensive therapy. PMID- 8221554 TI - Time trends of early gastric carcinoma. A clinicopathologic analysis of 2846 cases. AB - BACKGROUND: Despite recent improvements in the diagnostic accuracy and techniques for identifying early gastric carcinoma, gastric carcinoma remains a major cause of death in the world. METHODS: The clinicopathologic features of 2846 surgically resected early gastric carcinomas from 1962-1987 were reviewed, and the time trends of a variety of clinicopathologic factors were studied using both an univariate and a multivariate analysis. RESULTS: The differentiated type of carcinoma increased in frequency from 34-50%. The percentage of patients older than 60 years of age increased from 32-56%. The number of lesions measuring less than 3 cm increased from 28-56%. The frequency of shallow-type-IIc carcinomas measuring less than 3 cm, which had been previously difficult to detect and were mainly composed of differentiated carcinomas, increased from 46-69%. The relative proportion of the differentiated type of carcinoma was significantly higher in patients older than 60 years of age than in those younger than 60. The relative proportion of differentiated-type carcinomas measuring less than 3 cm, which had been easy to detect as opposed to undifferentiated-type carcinomas, significantly increased from 56-76%. CONCLUSIONS: The above time trends may indeed reflect genuine change. However, the recent increase in the rate of differentiated-type, small-sized carcinomas also suggests the possibility that such results may, in fact, be due mainly to improvements in both diagnostic accuracy and medical technology. Thus, a large number of undifferentiated-type, small-sized carcinomas might have merely failed to be found in routine gastric examination. PMID- 8221555 TI - Fluorodeoxyuridine with continuous leucovorin infusion. A phase II clinical trial in patients with metastatic colorectal cancer. AB - BACKGROUND: Chemotherapy of colon cancer has used the modulation of 5 fluorouracil (5-FU) by leucovorin; however, early studies indicate that leucovorin with fluorodeoxyuridine (FUDR) may be clinically superior. The authors report their experience in patients with metastatic colorectal cancer. METHODS: One hundred twelve evaluable patients with metastatic colorectal cancer were treated with leucovorin and FUDR. Leucovorin was given by continuous intravenous infusion at 500 mg/m2/day on days 1-6, and FUDR was given by intravenous push on days 2-6 at 3:00 p.m. daily, with doses ranging from 270-1350 mg/m2/day. RESULTS: This regimen was well tolerated with dose-limiting toxicity of diarrhea and stomatitis, while hematologic toxicity was minimal. At least one chemotherapy regimen had previously failed in 90 of 112 patients (80%). Twenty major responses greater than or equal to partial remission, lasting from 2-40+ months, were observed in this patient population for an overall response rate of 18%. Twelve of 22 previously untreated patients (55%) had major responses. Overall survival of previously untreated patients was 73% (14 of 19) and 50% (11 of 22) at 1 and 2 years, respectively. Of 66 patients who had received prior leucovorin-5-FU (LVFU) therapy with subsequent disease progression, 4 had major responses lasting 95, 241, 350, and 432 days, respectively. CONCLUSIONS: These data suggest that the modulation of FUDR by leucovorin may have clinical use. The recommended starting dose of FUDR is 800 mg/m2/day on days 2-6, with subsequent escalation each month in those patients who do not display stomatitis. PMID- 8221556 TI - Combined cystic teratoma and hepatoblastoma of the liver. Probable divergent differentiation of an uncommitted hepatic precursor cell. AB - BACKGROUND: Hepatic cystic teratoma is an extremely rare tumor and hepatoblastoma a relatively rare tumor of childhood. Their occurrence as a combined tumor has not been previously reported. METHODS: The relevant clinical, radiologic, and pathologic findings are presented. RESULTS: A 17-month-old boy presented with an abdominal mass. Imaging studies suggested an intrahepatic teratoma, but serum alpha-fetoprotein levels were extremely elevated. The resected tumor comprised adjoining benign cystic teratoma and epithelial hepatoblastoma. The child remains alive and well 37 months after complete resection of the tumor. CONCLUSIONS: The tumor was considered most likely to originate from divergent differentiation of a single uncommitted liver precursor cell. PMID- 8221557 TI - DNA analysis in hepatoblastoma by flow and image cytometry. AB - BACKGROUND: In several types of tumors, including hepatocellular carcinoma, prognosis could be correlated with DNA ploidy. Few studies have been performed on hepatoblastoma with contradictory results. METHODS: Twenty-nine cases of nonpretreated hepatoblastoma were studied with flow cytometry and image cytometry for DNA index and proliferation index using paraffin-embedded tissue. RESULTS: Twenty-three (79.9%) tumors were diploid, and 6 (20.7%) were aneuploid (hyperdiploid). Patients with diploid tumors were younger than those with aneuploid tumors. With regard to stage, diploid tumors were almost equally distributed among stages (tumor, lymph node metastases, distant metastases), whereas aneuploid tumors tended to occur in higher stages (tumor, lymph node metastases, distant metastases). Diploid tumors had clearly a better prognosis than aneuploid tumors, although the difference was not statistically significant (flow cytometry, P = 0.06; image cytometry, P = 0.16). A more favorable prognosis was also noted for hepatoblastomas with low-proliferation index (< or = 7%), but the difference from tumors with high-proliferation index (> 7%) again was not statistically significant (P = 0.16). CONCLUSIONS: Although no statistically significant differences in prognosis between hepatoblastomas with diploid and aneuploid DNA content, respectively, were found, there is a clear tendency that diploid hepatoblastomas behave more favorably. The same is true for hepatoblastomas with low-proliferation index. PMID- 8221558 TI - Heterogeneity in the nuclear DNA content of cells in carcinomas of the biliary tract and pancreas. AB - BACKGROUND: The authors investigated the frequency of aneuploidy and heterogeneity in DNA ploidy within one tumor by multiple sampling. METHODS: Using flow cytometry, the authors measured the nuclear DNA content of cells in fresh surgical specimens from 24 patients with carcinoma of the extrahepatic bile duct, 8 patients with carcinoma of the gallbladder, 5 patients with carcinoma of ampulla of Vater, and 14 patients with carcinoma of the pancreas. RESULTS: The frequency of aneuploidy was 91.7%, 87.5%, 80.0%, and 85.7% in carcinoma of the extrahepatic bile duct, gallbladder, ampulla of Vater, and pancreas, respectively. The incidence of heterogeneity in DNA ploidy within a tumor was 79.2%, 87.5%, 80.0%, and 85.7% in carcinoma of the extrahepatic bile duct, gallbladder, ampulla of Vater, and pancreas, respectively. The DNA index ranged mostly from 1.0-2.0. Hypodiploid samples constituted 23.5% of the total, and only five (9.8%) had a DNA content above tetraploid. Many of the patients in which the DNA index was about 1.0 (including diploid cases) tended to be in the relatively early stages of the disease. CONCLUSIONS: It is necessary to take several fresh samples from the same tumor to accurately measure the variations in nuclear DNA content. The authors suggest that three or four samples should be studied to determine aneuploidy and that DNA ploidy heterogeneity should be analyzed in five or six samples. PMID- 8221559 TI - Combination of chemotherapy and recombinant alpha-interferon in advanced non small cell lung cancer. Multicentric Randomized FONICAP Trial Report. The Italian Lung Cancer Task Force. AB - BACKGROUND: Preclinical data suggested that alpha-interferon (IFN) may potentiate chemotherapy cytotoxicity. METHODS: A prospective multicentric randomized trial was initiated to assess the clinical benefit of adding recombinant alpha-2-IFN to combination chemotherapy in patients with metastatic non-small cell lung cancer. A total of 182 patients were randomized to receive either cisplatin epidoxorubicin-cyclophosphamide (CEP) combination chemotherapy (cisplatin, 60 mg/m2; epidoxorubicin, 50 mg/m2; and cyclophosphamide, 400 mg/m2 intravenously) alone on day 1 or the same chemotherapy plus recombinant alpha-2-IFN at the dose of 5 MU intramuscularly from day -2 to +4, then 3 times weekly. RESULTS: The median survival was 6 months in the CEP plus IFN arm versus 5.5 months in the control arm. The log-rank test showed a marginal statistically significant difference (P = 0.045) in favor of CEP chemotherapy, which disappeared when survival curves were adjusted for prognostic factors. Progression-free survival was similar in the two treatment arms. Considering all eligible patients, the response rate was 7.6% in the CEP arm versus 18.9% in the CEP plus IFN arm (P = 0.042). Nearly 40% of the patients receiving IFN had grade 3-4 nadir leukopenia versus 15% in the control arm (P = 0.01) and 12.5% versus 4.2% had grade 3-4 thrombocytopenia. Apart from the usual constitutional symptoms, IFN was also responsible for increased emesis and mucositis. CONCLUSIONS: This study indicates that the addition of recombinant alpha-IFN to CEP chemotherapy can increase response rate and toxicity to treatment without a positive effect on progression free survival and survival. PMID- 8221560 TI - Mucin 1 antigens in the serum and bronchial lavage fluid of patients with lung cancer. AB - BACKGROUND: Lung cancer (LC) is the most common fatal malignancy, but there are no useful tumor markers for diagnosis or monitoring. Mucin 1 has an established role as a marker in other malignancies, but has undergone limited assessment in LC. METHODS: Serum from 86 patients with LC and 24 with benign pulmonary disease (BPD), and bronchial lavage fluid from 55 LC patients and 21 BPD patients were tested using the Mucin 1 assays mammary serum antigen (MSA) and cancer-associated serum antigen (CASA). RESULTS: For LC, serum CASA achieved sensitivity of 57%, specificity of 93% relative to normals, and 63% specificity relative to BPD. For MSA the same parameters were 19%, 95%, and 92%. Serum CASA levels were significantly higher in LC patients compared with BPD (P = 0.024) but there was no difference for MSA (P = 0.635). CASA showed excellent correlation with tumor stage and in patients with changing status of disease, while MSA did not. By contrast there was no difference in bronchial lavage fluid tumor marker levels from LC and BPD patients (CASA, P = 0.87; MSA, P = 0.89). CONCLUSIONS: In a small series serum CASA appears to be a useful agent in detecting LC because it is elevated in all types and stages of LC, and its level correlates with stage and progress of disease. Some patients with BPD have elevated levels suggesting a greater value for monitoring rather than diagnosis. Both serum MSA testing and measurements of either marker in bronchial lavage fluid are of no value. PMID- 8221561 TI - Expression and modulation of class I and class II histocompatibility leukocyte antigens on human soft tissue sarcomas. AB - BACKGROUND: Class I and Class II histocompatibility leukocyte antigens (HLA) play an important role in the antigenic recognition and target cell killing by T lymphocytes. Their expression and modulation with gamma interferon on human soft tissue sarcomas were investigated. METHODS: The phenotypic expressions of Class I and Class II HLA were determined by avidin-biotin immunoperoxidase staining using two monoclonal antibodies W6/32 and MEL3, respectively. RESULTS: The present study showed that soft tissue sarcomas frequently had demonstrable Class I HLA and less-frequently expressed Class II HLA: The staining for Class I HLA was more diffuse, and the staining for Class II HLA was generally patchy in appearance. The expressions of two antigens on cultured sarcoma cells were found in accordance with the findings of sarcoma tumors. The expression of Class I antigen was enhanced, and Class II was induced in two cell lines by gamma interferon. The in vitro modulation of HLA with gamma interferon was reversible. Gamma interferon at the testing dose did not have cytotoxic or antiproliferative effects on either cell lines. CONCLUSIONS: Through the modulation of HLA on soft tissue sarcomas, gamma interferon may play a role in the clinical management of sarcomas. PMID- 8221562 TI - Ambulatory continuous infusion ifosfamide with oral etoposide in advanced sarcomas. AB - BACKGROUND: Chemotherapy given by continuous infusion may have different toxicity profiles and different degrees of therapeutic efficacy than when given by bolus administration. The potential therapeutic benefits of continuous infusion chemotherapy and the advantages of outpatient treatment led us to study a continuous infusion of ifosfamide with mesna and oral etoposide. METHODS: The authors performed a Phase I-II trial in which 9 g/m2 ifosfamide was administered for 6 days and 10.5 g/m2 mesna was administered for 7 days, both by continuous infusion, in combination with 50 mg/m2/d oral etoposide for 8 days in 21 patients with sarcomas or other solid tumors. Courses were repeated every 28 days. RESULTS: A total of 65 treatment cycles were given. Only six patients required hospitalization for treatment, all because of an initial poor performance status, and most carried out normal activities on an ambulatory basis. Treatment was stopped during the first course in five patients because of central nervous system toxicity, each with a poor pretreatment performance status; neurologic recovery was complete in each patient. The dose of etoposide was decreased by 20% in 11 patients and unchanged in 7 following the first treatment. Hematologic toxicity was predominantly manifested by leukopenia. An absolute neutrophil count less than 500 neutrophils/microliters occurred in 22 of 50 cycles; thrombocytopenia (platelets less than 100,000/microliters) was seen in two patients, requiring platelet transfusion in one. Neutropenic fevers occurred in 13 of 65 cycles; in 4 of these, cultures demonstrated a bacterial infection. Nausea and vomiting were mild. Objective responses occurred in 6 of 16 patients with soft tissue sarcomas (6 partial responses [PR]) (95% confidence interval, 15 65%), and 3 of 5 bone sarcomas, all of whom had been previously treated with doxorubicin and dacarbazine. CONCLUSIONS: The authors concluded that ifosfamide and mesna given by ambulatory continuous intravenous infusion with wearable pump systems in combination with oral etoposide was well tolerated and showed substantial anti-tumor activity. This combination represents a rational therapeutic approach to patients with advanced soft tissue sarcomas and may have application to other malignancies. PMID- 8221563 TI - Short-term administration of granulocyte-macrophage colony stimulating factor decreases hematopoietic toxicity of cytostatic drugs. AB - BACKGROUND: Administration of granulocyte-macrophage colony stimulating factor (GM-CSF) is followed by a rapid increase in the proliferative activity of the hematopoietic precursors. Within 72 hours after its suspension, however, establishment of a negative feedback results in a reduction of the proliferative activity of the hyperplastic marrow to values below the baseline, suggesting refractoriness of hematopoietic progenitors to the action of cell-cycle-specific cytostatic agents. METHODS: The hypothesis that short treatment with GM-CSF before chemotherapy could reduce the hematopoietic toxicity of cytostatics was investigated by administering GM-CSF glycosylate (Sandoz, Basel, Switzerland/Schering-Plough, Kenilworth, NJ) subcutaneously with a 5.5 micrograms/kg protein dosage per day from day-6-day-4 before each course of adjuvant chemotherapy (cyclophosphamide, epirubicin, 5 fluorouracil/cyclophosphamide, methotrexate, 5-fluorouracil alternate) in patients with node-positive breast cancer. Twelve patients were randomized to receive GM-CSF before chemotherapy or only at chemotherapy. The hematologic picture and dose intensity of chemotherapy were compared in the two groups of patients. RESULTS: In the group of patients receiving chemotherapy only, 22% of the cycles had to be postponed because of leukopenia, with a consequent reduction of the dose intensity, whereas in the GM-CSF group, the neutrophil counts remained at significantly (P < 0.001) higher levels, and there were no delays in chemotherapy administration. No substantial systemic toxicity was associated with this brief GM-CSF schedule. Moreover, GM-CSF treatment did not result in delayed depletion of the hematopoietic pool. CONCLUSIONS: Short treatment with GM-CSF can enable the dose intensity of conventional protocols of proven efficacy to be increased. PMID- 8221564 TI - Race, income, and survival from breast cancer at two public hospitals. AB - BACKGROUND: Some studies have shown that adjustment for socioeconomic status reduces breast cancer survival differences between blacks and whites. The purpose of this study is to evaluate the effect of age, race, stage, treatment, and income status on breast cancer survival among women attending public hospitals in Chicago, Illinois. METHODS: Hospital Cancer Registry data on 887 black women and 265 white women with breast cancer onset between 1973-1985 were analyzed using Cox regression and Kaplan-Meier techniques. The purpose was to examine the effect of age, race, stage, treatment, and income on breast cancer survival. RESULTS: Black women with breast cancer were younger and poorer than white women with breast cancer. There were no significant differences between blacks and whites with regard to stage, estrogen receptor status, or type of treatment. Black women had lower 5-year breast cancer survival rates compared to white women (50.2% versus 60.2%; P = 0.05), and survival was lower when adjusted for stage and age. However, when adjusted for income in addition to stage and age, the effect of race on survival was reduced (from relative risk = 1.26; 95% confidence interval = 1.02, 1.57 to relative risk = 1.17%; 95% confidence interval = 0.95, 1.38). CONCLUSIONS: Income influences breast cancer survival differences between blacks and whites in this population. PMID- 8221565 TI - Overexpression of Glut-1 glucose transporter in human breast cancer. An immunohistochemical study. AB - BACKGROUND: Breast cancers have higher than normal glucose metabolism, but the mechanism of glucose entry into these tumors is not well understood. METHODS: The expression of five facilitative glucose transporters, Glut-1 (erythrocyte type), Glut-2 (liver type), Glut-3 (brain type), Glut-4 (muscle/fat type), and Glut-5 (small intestine type), was studied by immunohistochemistry of paraffin sections from 12 primary human breast cancers and 8 lymph node metastases from 2 patients. Rat tissues known to express these glucose transporters were used as controls. RESULTS: All the primary breast cancers and the lymph node metastases were positive for Glut-1. This transporter was expressed on the cell membrane and in the cytoplasm of the tumor cells, but exhibited marked intratumoral and intertumoral variability in the proportions of positive cells and the intensity of staining. Staining of the normal mammary epithelium, if present, was much lower than observed in tumor cells from the same patient. Glut-2 was expressed in all of the tumors, but the intensity of staining was not consistently stronger than that seen in healthy breast. Clusters of Glut-4-positive granule were observed in cells in six of the tumors. None of the tumors or the healthy breast in the tissues studied expressed Glut-3 or Glut-5. CONCLUSIONS: Higher expression of the glucose transporter Glut-1 by breast cancer cells compared with the healthy breast tissue is common. Increased glucose transporter protein expression may contribute to the increased uptake of 2-[18F]-fluoro-2-deoxy-D-glucose (FDG) by these tumors observed by positron emission tomography (PET) imaging. PMID- 8221566 TI - Response to the National Cancer Institute Alert. The effect of practice guidelines on two hospitals in the same medical community. AB - BACKGROUND: Despite the recent increase in medical practice guideline development and dissemination, physician compliance with the guidelines has often been low. Previous research has suggested that physicians at hospitals with low volumes of cases and weakened financial status were more likely to omit indicated diagnostic testing or appropriate treatment. The authors sought to determine whether differences in compliance to a widely disseminated set of guidelines would exist even among the most dominant hospital providers within the same medical community. METHODS: Two hospitals, together providing nearly half of the cancer surgery within a metropolitan area, were studied for their compliance to the May 1988 National Cancer Institute (NCI) Clinical Alert regarding adjuvant therapy after primary treatment for node negative breast cancer. A case series consecutive collection of 549 women treated at the study hospitals for 2 years before and two years after the Alert determined those patients who had received any form or combination of adjuvant therapy after primary surgical treatment (lumpectomy or modified radical mastectomy). RESULTS: Following modified radical mastectomy, for women age 50 and older, the university hospital (U) provided adjuvant therapy to a higher percentage of patients than the community hospital (C) both before (25.6% versus 4.7%, P < 0.005) and after (58.9% versus 23.2%, P < 0.001) the Alert. For women younger than 50 years of age, the two hospitals were equally likely to provide adjuvant therapy both before and after the Alert. Following lumpectomy, hospital U increased the percentage of women receiving adjuvant therapy following the Alert in women younger than 50 years of age (25 75.8%, P < 0.001) and in women age 50 and older (33.3-56.5%, P < 0.025). Hospital C provided no adjuvant therapy before or after the Alert. Preferences for breast conserving surgical treatment were significantly (P < 0.001) different with hospital U performing a higher percentage of lumpectomies than hospital C both before (50.9% versus 14.9%) and after (57.6% versus 16.8%) the Alert. CONCLUSIONS: Significant differences in compliance with practice guidelines may be found even among the most dominant hospital providers of cancer services within the same medical community. The role of the surgeon in referring patients to the oncologist greatly influences the ultimate provision of adjuvant therapy. Strategies for enhancing compliance should be considered integral to the process of guideline development. PMID- 8221567 TI - Time dependency of the influence of prognostic factors on relapse in breast cancer. AB - BACKGROUND: A number of factors concerning the prognosis of cancer patients have been reported, but the evaluation of individual factors is not always consistent. The author considered that the discrepancies may lie in part in that factors may have a changing influence on relapse with time. The present study was intended to reveal the phenomenon and its clinical significance in patients with breast cancer. METHODS: Eleven pathological prognostic factors in 462 patients with primary breast cancer were analysed with special reference to the influence on relapse and its changes with time. The Cox regression analysis was performed for whole and three stratified non-overlapping observation period (within 2, 2-5 and 5 years more), using factors including extent of nodal involvement (N), nuclear grade (NG), tumor infiltrating lymphocytes (TIL), tumor size, fat infiltration (F) and others. RESULTS: Cox regression analysis for whole observation period showed N followed by NG, TIL, F and others as the most important factors on relapse. But the analysis performed for 3 stratified intervals revealed that the influence on relapse decreased with time for N and NG, but tended to increase with time for F. The suppressive effect of TIL on relapse was not evident up to 2 years but was significantly emphasized after 2 years postoperatively. CONCLUSIONS: In conclusion, a distinct time-dependency with varied patterns was found in the influence of some prognostic factors on relapse, which implies the assessment of prognostic factors may vary according to the length of the observation period. PMID- 8221568 TI - Cervical cytopathology. An evaluation of its accuracy based on cytohistologic comparison. AB - BACKGROUND: Although Papanicolaou cytology represents the most effective technique to prevent and detect precancerous conditions of the uterine cervix, its false-negative yield is still a reason of concern among pathologists and gynecologists. METHODS: Because histologic control is one of the best ways to assess the accuracy of cytology diagnosis, the authors have investigated 1000 women who had cervical smears and tissue sampling obtained during the same colposcopic evaluation between 1987 and 1990. RESULTS: Out of 1000 cases (average age, 34.6 years; range, 14-80 years), 918 had adequate, 62 had less than optimal, and 10 had unsatisfactory samples. Cytology unsatisfactory and less than optimal cases as well as inadequate histology cases have been disregarded from all calculations. After histologic comparison, confirmed negatives were 622 of 918 (67.8%). Cytologic diagnoses of cervical intra-epithelial neoplasia (CIN) I were 96, of CIN II were 44, of CIN III, inclusive of carcinoma in situ, were 39, and of invasive carcinoma were 2. Atypical cases were 56. The overall sensitivity was 76.3%, with group sensitivity rates increasing directly with CIN grade. Positive predictive value was 80.2%. Specificity was 93.0%, and negative predictive value was 91.3%. False-negatives were 59 of 681 (8.7%), basically due to sampling errors. Among true-positives, there was 1 category discrepancy in 30 cases (mostly undercalled or overcalled CIN II) and 2 category discrepancies in 4 cases. CONCLUSIONS: Cervical cytology has an overall accuracy close to that reported in studies employing indirect control methods, such as patient follow up. Higher sensitivity rates emerged for CIN II, CIN III, and cervical carcinoma. Our figures of sensitivity and specificity may represent a useful reference source for future studies dealing with quality control in cervical cytopathology. PMID- 8221569 TI - National survey of ovarian carcinoma. VI. Critical assessment of current International Federation of Gynecology and Obstetrics staging system. AB - BACKGROUND: The Commission on Cancer of the American College of Surgeons recently completed a national survey of patients with ovarian cancer. From the large database, the prognostic value of current International Federation of Gynecology and Obstetrics (FIGO) staging system for ovarian carcinoma was re-examined. METHODS: Data was collected from 25 consecutive ovarian carcinomas diagnosed in 1983 and 1988 at 904 hospitals with cancer programs. Among a total of 12,316 cases, 5156 patients had long-term survival data. RESULTS: The overall 5-year survivals were 88.9 +/- 0.9%; 57.1 +/- 2.4%; 23.8 +/- 1.3%; and 11.6 +/- 0.9% for Stages I, II, III, and IV, respectively. Pairwise survival comparisons using Lee Desu statistic confirmed the prognostic value of current staging system (P < 0.00001). When survival data was substratified further to substage division, the 5-year survivals were: IA, 92.1 +/- 0.9%; IB, 84.9 +/- 3.4%; IC, 82.4 +/- 2.0%; IIA, 69.0 +/- 4.3%; IIB, 56.4 +/- 3.6%; IIC, 51.4 +/- 4.5%; IIIA, 39.3 +/- 2.8%; IIIB, 25.5 +/- 2.6%; IIIC, 17.1 +/- 1.4%; and IV, 11.6 +/- 0.9%. As the disease process becomes more advanced, patients' survival reduces proportionally. However, the survival reduction is relatively small between IB-IC and IIB-IIC divisions. Survival comparisons revealed significant prognostic value for most substage divisions (P = 0.03-0.0002) except for IB-IC and IIB-IIC combinations (P > 0.33). Further analyses revealed no significant differences between IB-IC and IIB-IIC patients in several prognostic parameters such as age, histologic grade, cell type, and amount of residual disease. CONCLUSIONS: These data support the current FIGO staging system. However, Substages IB-IC and IIB-IIC should be combined to respective single substages. PMID- 8221570 TI - Metallothionein in testicular germ cell tumors and drug resistance. Clinical correlation. AB - BACKGROUND: Metallothioneins (MT) are endogenous metalloproteins involved in the homeostasis of essential metals and detoxification of toxic metals. Some recent experimental studies suggested tumor resistance to cisdiamminedichloroplatin may be associated with overexpression of MT in the tumor. METHODS: The presence of MT in 33 primary testicular germ cell tumor specimens was assessed immunohistochemically using a rabbit polyclonal rat liver MT antibody that cross reacted with human MT. The data were correlated with the patients' clinical course. RESULTS: Seminomas stained weakly or not at all for MT, regardless of the clinical stage. Most nonseminomas stained heavily for MT. The more advanced staged nonseminomas tended to stain more heavily for MT. CONCLUSIONS: In view of the considerable experimental evidence as well as some inferential clinical data involving MT in cis-diamminedichloroplatin resistance, there appears to be a role for MT in cis-diamminedichloroplatin resistance in germ cell tumors. Further studies to elucidate the role of MT in germ cell tumor chemoresistance are warranted. PMID- 8221571 TI - Bladder cancer definitive radiation therapy of muscle-invasive bladder cancer. A retrospective analysis of 317 patients. AB - BACKGROUND: The role of radiation therapy as curative treatment of muscle invasive bladder cancer was to be analyzed. METHODS: From 1980-1990, 308 patients with transitional cell carcinoma of the urinary bladder received definitive pelvic radiation therapy (nominal standardized dose greater than or equal to 1700 ret). T categorization was based on clinical examination assessing the palpability of the bladder tumor and its extent (TNM 1978/1982). RESULTS: The cancer-specific 5-year survival rate for all patients was 24% (crude survival, 20%). The 135 patients with T2/T3a tumors lived significantly longer (5-year survival, 38%) than those with greater than or equal to T3b tumors (5-year survival, 14%). In the former group of patients, age (75 years and younger versus older than 75 years) was significantly correlated with a favorable outcome. The cancer-specific 2-year survival was significantly correlated to clinical response assessed 3-4 months after radiation therapy was 72%, 38%, and 10% in cases of complete response, partial response, and no response/inevaluability, respectively. In a multivariate analysis, the T categorization, patient age, serum creatinine level (less than or equal to 150 mumol/l versus greater than 150 mumol/l), and radiation therapy schedule predicted the 5-year survival rate. CONCLUSIONS: The clinical T category (< or = T3a versus > or = T3b), based on bimanual palpation, represents an important prognostic parameter, if done by clinicians experienced in onco-urology. High-dose radiation therapy offers a reasonable chance for long-term survival in patients with T2/T3 tumors confined to the bladder wall, especially in patients younger than 76 years. Greater than or equal to 80% of patients with more extended tumors (greater than or equal to T3b) and those older than 75 years of age are not curable by radiation therapy alone. In these patients palliative treatment modalities should be considered, in particular if cisplatin-based chemotherapy is not feasible. PMID- 8221572 TI - A bladder cancer multi-institutional experience with total cystectomy for muscle invasive bladder cancer. AB - BACKGROUND: The role of total cystectomy was to be assessed in the curative treatment of muscle-invasive bladder cancer. METHODS: Two hundred and fifty-three patients with T2-T4a transitional cell carcinoma of the urinary bladder were referred to precystectomy radiation therapy (46 Gy, 66 patients; 20 Gy, 187 patients). These patients represented approximately 20% of all patients developing muscle-invasive bladder cancer in Southern Norway from 1980-1990. The clinical T categorization was generally based on palpability and extent of the palpable bladder tumor assessed by the referring urologist. Twenty-six patients (10%) did not have total cystectomy, most often due to peroperatively demonstrated locoregional inoperability. Two or three cycles of cisplatin-based combination chemotherapy were given to 68 patients. RESULTS: For the 227 patients who underwent cystectomy, the cancer-specific 5-year survival rate was 58% (T2 [104 patients], 63%; greater than or equal to T3 [123 patients], 54%) (P = 0.022). The comparable figure for patients with histologically proven regional lymph node metastases was 22%. The 97 stage-reduced cases (less than or equal to pT1) survived significantly longer than the 130 patients without stage reduction (74% versus 46%) (P < 0.0001). Neoadjuvant chemotherapy was correlated with a more favorable survival in patients with greater than or equal to T3 tumors but did not seem to influence survival of patients with T2 bladder cancer. CONCLUSIONS: In a multicenter setting, prognostically relevant T categorization of operable muscle-infiltrating bladder cancer can be based on the palpability of the primary tumor. Approximately 50% of favorably selected patients with operable T2-T4 bladder cancer survived for at least 5 years independent of whether the operation was done at a large uro-oncologic unit or a smaller urologic section. In this retrospective review, chemotherapy seemed to improve the survival in patients with deeply infiltrating (greater than or equal to T3) bladder cancer but appeared to represent an overtreatment in patients with T2 tumors. PMID- 8221573 TI - Regional heterogeneity in the DNA content of human gliomas. AB - BACKGROUND: Regional heterogeneity causes significant errors in the histologic classification and grading of gliomas, but little is known about its implications for other modalities. Attempts to predict glioma behavior using flow cytometry (FCM) have yielded contradictory results, possibly due to regional heterogeneity in DNA content, a recognized phenomenon that has not been evaluated systematically. METHODS: The authors used FCM to analyze the DNA content of 353 regions from 18 resected human gliomas. Five to 60 regions were sampled from each tumor, and the topographic relationships of ploidy, proliferative activity (S phase fraction [SPF]), and histologic features were established. RESULTS: Most tumors demonstrated heterogeneity among regions in the number and relative sizes of aneuploid populations and/or proliferative activity. The degree of heterogeneity increased with tumor grade. Heterogeneity in histologic features, ploidy, and proliferative activity appeared to vary independently, except for a significant association between the presence of mitotic figures and the SPF of individual regions (P < 0.0001). The clustering of regions with similar percentages of aneuploid cells supported the concept of local clonal expansion. CONCLUSIONS: Gliomas express significant regional heterogeneity in ploidy and proliferative activity and this will have an adverse effect on the usefulness of their analysis. However, the independent variation of ploidy, proliferative activity, and histologic features suggests that the use of multiple analyses may allow more accurate prediction of glioma patient survival. Regional heterogeneity appears be a fundamental property of malignant gliomas; systematic studies to determine its effects on the diagnostic usefulness of other new methods used to evaluate gliomas are indicated. PMID- 8221574 TI - Endothelial cell marker PAL-E reactivity in brain tumor, developing brain, and brain disease. AB - BACKGROUND: The endothelial cell marker PAL-E is not reactive to vessels in the normal brain. The present study concerns the PAL-E reactivity in brain tumors in contrast to normal brain and nonneoplastic brain disease. METHODS: A total of 122 specimens were examined: brain tumors (n = 94), nonneoplastic brain disease (n = 19), normal brain (n = 8), and fetal brain (n = 1). Standard immunohistochemical procedures using a panel of endothelial cell markers were applied to detect vessels reactive to PAL-E. RESULTS: PAL-E reactivity to endothelial cells was found in all cases of glioblastoma multiforme, in 75% of the cases of anaplastic astrocytoma, and in 46% of the cases of astrocytoma. Furthermore, PAL-E reactivity was present in diseases with a developmental etiology, such as primitive tumors and congenital vascular malformations. The developing human brain (6-weeks' gestation age) and special sites of the mature brain, sites without blood-brain barrier, showed a strong reactivity, which indicates a relation with the status of blood-brain barrier development. CONCLUSIONS: PAL-E is the only marker out of a panel of endothelial cell markers that shows no reactivity to endothelial cells in the normal brain with an intact blood-brain barrier. In primary and metastatic brain tumors, PAL-E is reactive to endothelial cells, except for 25% of anaplastic astrocytoma and 54% of astrocytoma. PAL-E reactivity in brain tumors most likely is related to angiogenesis and to blood tumor barrier properties not present in the normal blood-brain barrier. PMID- 8221575 TI - CD30 (Ki-1) positive anaplastic large cell lymphomas in individuals infected with the human immunodeficiency virus. AB - BACKGROUND: CD30 (Ki-1) positive anaplastic large cell lymphoma (ALCL) has been only rarely described in HIV-positive patients. METHODS: The clinicopathologic features of eight ALCLs occurring in four AIDS and four HIV-positive patients were investigated. The phenotype of each neoplasm was determined by immunohistochemical methods. In three cases fresh tissue was available for molecular analysis. RESULTS: The ALCLs are a clinically heterogeneous group of T (4), B (1) and indeterminate (3) cell malignant lymphomas which presented in the skin (4), liver (1), lung (1), nasal cavity (1; also with bone marrow involvement) and peritoneal fluid (1). While most of the patients had aggressive disease, dying in a median of three months, two patients had either localized or regressing skin lesions. Molecular studies showed that two ALCLs, one of B cell and one of indeterminate cell lineage, contained clonal Epstein-Barr virus sequences. None of the ALCLs examined contained evidence of HTLV-1 or HIV integration nor did they exhibit c-myc or bcl-2 proto-oncogene rearrangements. No mutations or deletions of the p53 tumor suppressor gene were identified in the three cases studied. CONCLUSIONS: HIV-related ALCL represents a clinically heterogeneous group of T cell, B cell and null cell malignant lymphomas, distinct from the previously described categories of AIDS-associated NHL, that may expand the spectrum of lymphoid neoplasms associated with HIV-infection. Identification and investigation of other cases of HIV-associated ALCL is important to determine the nature of the relationship between HIV infection and the development of ALCL. PMID- 8221576 TI - Prognostic factors in patients with metastatic malignant melanoma. A multivariate analysis. AB - BACKGROUND: Current methods to predict survival in patients with advanced, metastatic melanoma are limited. To determine clinical prognostic factors that accurately predict survival in patients with metastatic melanoma, a retrospective analysis was performed. METHODS: Clinical, hematologic, and biochemical pretreatment parameters from 284 patients entered on 18 Phase I and II clinical trials were analyzed to determine their prognostic effect on survival. A multivariate parametric regression based on the Weibull distribution was derived to estimate survival. RESULTS: Multivariate Weibull survival regression analysis showed that a simple model using the logarithm of the pretreatment values of lactate dehydrogenase and serum albumin, dichotomized as high and low, significantly and adequately predicted survival. A more complex multivariate model was also derived that involved the pretreatment platelet count, visceral organ involvement, and gender as additional factors. However, a larger study is needed to statistically validate such a model. CONCLUSIONS: The pretreatment values of serum lactate dehydrogenase and albumin are independent prognostic factors for survival in patients with metastatic melanoma. These two factors can be used to estimate survival of patients with advanced, metastatic melanoma and should be considered when designing melanoma trials in which survival is an endpoint. PMID- 8221577 TI - Different contribution of substrates oxidation to insulin resistance in malnourished elderly patients with cancer. AB - BACKGROUND: The relative contribution of malnutrition and cancer to insulin resistance in elderly patients is still poorly understood. METHODS: Twenty malnourished and 15 well-nourished patients with cancer matched for age (70 +/- 0.3 versus 69 +/- 0.3 years; P = not significant), mean arterial blood pressure (104 +/- 8 versus 107 +/- 7 mm Hg; P = not significant), gender ratio (10 males/10 females versus 7 males/8 females), and sedentary life style underwent an euglycemic glucose clamp. Simultaneous infusion of 3-D-H-glucose and indirect calorimetry allowed the authors to determine glucose turnover parameters and analyze substrates oxidation. RESULTS: Malnourished patients displayed a reduced total body glucose metabolism (36 +/- 2 versus 31 +/- 3 mumol/kg of lean body mass x min P < 0.04) despite higher glucose oxidation. In the same group of patients, the authors demonstrated a significant increase in lipid oxidation and the lack of the inverse relationship between glucose and lipid oxidation (r = 0.07; P = not significant). Insulin infusion enhanced glucose oxidation and suppressed lipid oxidation, but such changes were at lower extent in malnourished patients. CONCLUSIONS: Our study confirms the presence of insulin resistance in malnourished patients and demonstrates the relative contribution of substrates oxidation to the insulin resistance itself. PMID- 8221578 TI - Liposarcoma in patients younger than or equal to 22 years of age. AB - BACKGROUND: There have been few reported series of liposarcomas in patients younger than or equal to 22 years of age. METHODS: A retrospective analysis of all patients presenting with liposarcoma between 1949-1990 at Memorial Sloan Kettering Cancer Center with age at diagnosis younger than or equal to 22 years was performed. Variables evaluated for their predictive effect on survival included anatomic location of the primary, size, and completeness of surgical resection. RESULTS: Eighteen patients were identified. Only 1 patient (6%) presented with a high-grade lesion, and in 13 patients (72%), the myxoid subtype was observed. All but one patient undergoing complete resection remain disease free 1.3-29.1 years after treatment, while all patients with gross residual tumor have died from disease. Two of three patients with microscopic residual at resection are disease-free 2 and 11.8 years after diagnosis with the addition of external beam radiation therapy. CONCLUSIONS: The authors conclude that complete surgical resection is crucial for survival in young patients with liposarcoma and the external beam radiation therapy may be effective against microscopic residual. PMID- 8221579 TI - Development of a new intensive therapy for acute lymphoblastic leukemia in children at increased risk of early relapse. The Memorial Sloan-Kettering-New York-II protocol. AB - BACKGROUND: Improved survival of children with acute lymphoblastic leukemia (ALL) has made it more difficult to develop new protocols to further improve results. The authors report the pilot experience with the Memorial Sloan-Kettering-New York-II (MSK-NY-II) protocol, based on the New York regimen with changes made in an attempt to improve efficacy while reducing toxicity. METHODS: Forty-four of 46 consecutive patients were randomized to one of four regimens varying only in the sequence and mode of administration of the drugs during the first 48 hours of therapy, while the kinetics of the disappearance of the leukemic cells from the bone marrow was monitored with bone marrow aspirates and biopsies on days 0, 2, 7, and 14. RESULTS: Thirty-two high-risk and 12 average-risk patients were randomized. The marrow contained less than 25% blasts in 74.4% and 92.9% by day 7 and 14, respectively. Ninety-three percent achieved remission. Regimens beginning with daunorubicin achieved a greater and more rapid reduction in leukemic cells than those starting with cyclophosphamide. Daunorubicin infusion produced a more rapid cytoreduction than daunorubicin bolus. Two of 41 patients who achieved remission relapsed, and there was one death in remission. With a median follow-up of 54+ months, the event-free survival (EFS) rate was 86% +/- 10%. Disease-free survival (DFS) rate at 48 months was 93%. The estimated 4-year EFS rate for the high-risk and average-risk patients were 83 +/- 14% and 93 +/- 10%, respectively. Four of 18 patients given daunorubicin bolus and 0 of 18 patients given daunorubicin infusion who were monitored with serial echocardiograms had significant decrease in cardiac function (P = 0.10). The major toxicity of the therapy was infections, with 35% of patients developing serious infections during induction and consolidation. Half the patients had an episode of bacteremia from the venous catheter during the 2 years of maintenance. CONCLUSIONS: Close monitoring of kinetics of cytoreduction can rapidly distinguish between similar therapies, and the surrogate end-point may reduce the need for the long follow-up periods that may still be required to demonstrate differences in EFS. Continuous infusion of daunorubicin had less cardiotoxicity with faster antileukemic activity than bolus infusion. The MSK-NY-II protocol with a 86% 4-year EFS rate and a 95% DFS rate was a promising new regimen for the treatment of average-risk and high-risk ALL. PMID- 8221580 TI - The American Joint Committee on Cancer. Criteria for prognostic factors and for an enhanced prognostic system. AB - The American Joint Committee on Cancer proposes the following criteria for evaluating putative prognostic factors: they must be (1) significant, (2) independent, and (3) clinically important. Furthermore, we suggest the criteria for selecting a prognostic system that includes TNM and new prognostic factors. These criteria are: (1) easy for physicians to use; (2) provides predictions for all types of cancer; (3) provides the most accurate relapse and survival predictions at diagnosis and for every year lived for each patient; (4) provides group survival curves, where the grouping can be by any variable including outcome and therapy; (5) accommodates missing data and censored patients and is tolerant of noisy and biased data; (6) makes no a priori assumptions regarding the type of data, the distribution of the variables, or the relationships among the variables; (7) can test putative prognostic factors for significance, independence, and clinical importance; (8) accommodates treatment information in the evaluation of prognostic factors; (9) accommodates new putative prognostic factors without changing the model; (10) accommodates emerging diagnostic techniques; (11) provides information regarding the importance of each predictive variable; and (12) is automatic. PMID- 8221581 TI - Tap in on ear thermometry. AB - In February 1992, nursing departments at The Mississauga Hospital, a 530-bed community hospital in Southern Ontario, purchased First Temp Genius ear thermometers to replace the electronic predictive thermometers then in use. Ear thermometers collect infrared heat radiation from the ear canal with a probe, then convert the radiation to a temperature reading. Although quick and easy to use with patients of all ages, there are nonetheless some issues concerning their use that still must be addressed. PMID- 8221582 TI - Self-governed health. AB - As primary caregivers in remote Indian communities in northern Canada, nurses often witness considerable health status inequities between these people and the rest of Canada's population. In Saskatchewan, for example, tuberculosis incidence rates are 15 times higher for northerners in general than for people in the south. Infant mortality rates are twice as high, suicides are three times higher, motor vehicle related deaths are four to six times higher and deaths from diabetes in Indians aged 45 to 65 years are six times higher than in the general population. Some people suggest that these discrepancies are due simply to the inadequate funding of health care programs. Others refer to the remoteness of the communities and hence the inability to access advanced medical technology. A final hypothesis is that the increased exposure to white communities, practices and morals and the neglect of spiritual values have led to social disintegration and have hindered good health care practices. PMID- 8221583 TI - Culture shock. AB - As Canadians, we pride ourselves on our tolerance toward diverse ethnic cultures within our country. Unfortunately, tolerance is not enough. Native and ethnocultural groups have unique health care needs, needs we do not always meet. Our health care system was established before the increase in non-European immigrants and without input from native groups. As a result, the values of many nurses still reflect those of the dominant culture, often to the exclusion of other cultures. Hopefully by understanding the differences in cultural values we can improve the health status of ethnocultural and native groups in Canada. PMID- 8221584 TI - Labrador health care comes home. AB - The recruitment and retention of nurses in isolated communities in Canada's North is a chronic problem. Non-native nurses, usually from urban areas, face cultural and lifestyle differences that make many unwilling to stay in these communities. Plus, they rarely have cross-cultural training or experience. On the other side, the residents must often discuss the private details of their lives with nurses from a foreign culture and in a language other than their native tongue. PMID- 8221585 TI - Crisis, grief and loss. AB - At one time or another, many of us experience a life-threatening crisis that proves to be a turning point in our lives. I had such a crisis while working as a medic on the oil-rig Vinland, offshore of Nova Scotia, in 1984. PMID- 8221586 TI - [Resources for preventing professional burnout]. AB - Burnout in the nursing profession has been reported in the literature for the past decade. Yet few studies have identified the causative factors or the recommended resources for prevention and treatment. An exhaustive literature review by the authors produced over 300 articles--40 of which were selected and systematically analyzed. It became apparent that the contributing factors to nursing burnout include stressors relating to the nurse's role, tasks and interpersonal conflicts, and in particular, the lack of management and peer support. The nurse's susceptibility to burnout varied considerably and was dependent upon individual character traits such as hardiness. Hardy individuals demonstrated an ability to deal with life through qualities such as their ability to manage change; their sense of commitment, and their sense of control over their lives. According to the literature, these individuals are less susceptible to burnout manifestations. The article also identifies potential resources available to nurses and positive factors that could improve their adaptation to stressful situations or environments. In light of dwindling financial resources and increased nursing responsibilities, the authors advise steering nurses in a direction that will unite them with their peers, management, and their profession. PMID- 8221588 TI - Documenting by computer. PMID- 8221587 TI - [Work satisfaction of psychiatric nurses]. AB - A demanding clientele, the social stigma of working in a psychiatric institution and a lack of professional support are factors that discourage psychiatric nurses from staying in their profession. Because of this, there is the potential risk of decreasing nursing's contribution to psychiatric treatment and rehabilitation- thereby compromising the quality of health care. In light of this, a research study was carried out to look at job satisfaction of psychiatric nurses in four Quebec hospitals. The study aimed to identify the direct repercussions on nurses caring for hospitalized psychiatric patients. The results of the study indicate that both staff nurses and nurse managers alike suffer a marked sense of job dissatisfaction. The key element is the apparent prioritization of administrative functions above clinical functions. The study recommended that necessary adjustments be made to create a favorable milieu that stimulates rehabilitation. The authors propose to enhance professional competence by making less bureaucratic demands on nurses; increasing the value of direct patient care; increasing professional supervision; providing more inservice education, and diminishing the hierarchial structure--thereby favoring clinical nursing practice. PMID- 8221589 TI - Reallocating power. AB - Nursing management throughout the country must accept the realization that the "profession" of nursing is at a crossroads. Nursing must either abandon its claim as a profession or revise nursing's support structures in order to provide a professional practice environment. Those who accept nursing as a profession understand the responsibility and authority of the profession. These values involve the desire to assume accountability, articulate the practice of nursing and nursing competence, and define the quality of care provided. As the majority of nurses continue to practice within the hospital setting, the challenge facing nursing management is the implementation of professional governance practices. PMID- 8221590 TI - [Nursing care evolves? That's to be seen!]. AB - This author sees a gap between nursing education and practice. She maintains that critical thinking and analysis of a nursing situation are part of the autonomous functions of a professional nurse. She also believes that nurses at the masters level should not only teach patients and their families, but also new nursing graduates in order to give them an inservice preparation to critical thinking. The author also suggests that new grads receive a supervised internship. PMID- 8221591 TI - Saying goodbye. PMID- 8221592 TI - A uniform with uniformity. AB - Clothing is a powerful form of non-verbal communication, conveying socio-economic status, occupational role and even the authority and power vested in that role. The traditional hospital nursing uniform reflects its historic military and religious foundations, and, many would say, the servile rank of the wearer. PMID- 8221593 TI - Under a grueling hot sun. PMID- 8221594 TI - Constraints on the moral agency of nurses. AB - Awareness of nurses' ethical concerns has increased greatly over the last few years. In response to this awareness, the Canadian Nurses Association revised the Code of Ethics to provide direction for the implementation of ethical standards for nursing. Yet, a growing body of literature suggests that situational constraints make it difficult for nurses to uphold these standards. Overcoming these constraints will take concerted efforts in nursing education, administration, practice and research. PMID- 8221595 TI - ECMO: some infants' last chance. PMID- 8221596 TI - Getting to your point. PMID- 8221597 TI - Calling the shots. PMID- 8221598 TI - Baby friendly care. AB - In 1991, the United Nations Children's Fund (UNICEF) and the World Health Organization (WHO) launched a global campaign called the Baby-Friendly Hospital Initiative (BFHI). The initiative encourages health professionals to promote, protect and support breastfeeding as the optimal method of infant nutrition and enables families to make an informed choice about infant feeding. Hospitals that serve perinatal families are the focus, because what starts in the hospital carries on into the community. Recognizing the vital role of nurses as patient advocates and health promoters, and their position as the largest group of hospital-based health care workers, WHO and UNICEF are asking nurses to facilitate the implementation of the initiative in their hospitals. PMID- 8221599 TI - Breastfeeding: then and now. AB - Over the centuries, civilization has survived because babies were breastfed. Although there has always been a small percentage of women (less than four per cent) unable to provide sufficient milk for their infants, the majority of women can produce enough milk to nourish twins and even triplets. PMID- 8221600 TI - [Nursing research]. AB - Research in the health sciences is a continuously evolving activity. Nursing research, however, is unfamiliar to many health professionals. Even nurses are sometimes uncomfortable with this activity. The goal of this article is to demystify nursing research. The authors define nursing and nursing research, its goals, history and the actual state of nursing research in Canada. PMID- 8221601 TI - [Mental health and support of women treated for breast cancer]. AB - This article presents the results of a pilot-study conducted on a small sample of women with breast cancer and the members of their social support system. The purpose was to determine the potential link between the mental health of the breast cancer patient and their support system. Mental health was assessed with the SCL-90-R (Derogatis, 1977), a self-report scale composed of 90 items divided into nine categories, offering three global indicators of distress: the Global Severity Index (GSI); the Positive Symptom Distress Index (PSDI), and the Positive Symptom Total (PST). The SCL-90-R was distributed to a sample of 14 subjects: seven women treated for breast cancer and seven significant others (social support). The results led to the definition of symptoms such as hostility, phobic anxiety and psychosis specific to either the woman or the support system. PMID- 8221602 TI - What is informed consent? PMID- 8221603 TI - It's all in the name. AB - In recent years it has become fashionable to call a person seeking health care a client instead of a patient. Some consider that this is part of the change in nursing, from the old military model that Florence Nightingale was acquainted with to the new consumer model. Nurses are also finding themselves in the position of acting as advocates for the care-seeking person. For me, none of these models appear appropriate for nursing. PMID- 8221604 TI - My baby's dying! PMID- 8221605 TI - Immunophenotype of mitotic cells with clonal chromosome abnormalities demonstrating multilineage involvement in acute myeloid leukemia. AB - Eleven patients with acute myeloid leukemia (AML) were studied with a technique that simultaneously identifies cytogenetic abnormality and immunophenotype of the same mitotic cell. To determine the cell lineages with abnormal karyotypes, monoclonal antibodies in the alkaline phosphatase-antialkaline phosphatase (APAAP) detection procedure were used. The granulocytic/monocytic lineage was involved in the leukemic process in all 11 patients. In nine patients, we also detected abnormal karyotypes in the erythrocytic and/or megakaryocytic lineages. All four patients with secondary AML showed involvement of the granulocytic/monocytic, erythrocytic, and megakaryocytic lineages into the leukemic process, as compared with five of seven patients with de novo AML. One patient with trisomy 8 showed erythrocytic participation in the leukemic process, but in another the erythrocytic lineage had only normal karyotypes. Thus, in AML, the chromosome abnormalities apparently usually originate at the multipotent progenitor cell stage, since in addition to granulocytic/monocytic lineages, erythrocytic and/or megakaryocytic lineages were also involved. Some patients show involvement of granulocytic/monocytic lineages only, however, suggesting that the target cell belongs to a more mature committed progenitor cell stage. PMID- 8221606 TI - Cytogenetic abnormalities and clinical stage in testicular nonseminomatous germ cell tumors. AB - To study the impact of chromosomal abnormalities on the clinical behavior of testicular nonseminomatous germ cell tumors (TNSGCTs), we compared the chromosomal constitution of primary tumors of patients who initially presented and remained without metastases to those with metastatic disease. Furthermore, the chromosomal pattern of primary TNSGCTs was compared to ploidy and the clinicopathologic risk factors histology and small-vessel invasion. The modal chromosome number and the ploidy were in agreement. No correlation was found between the modal chromosome number and histology, presence of vascular invasion, or clinical stage. No correlation was found between structural chromosome abnormalities, like the number of copies of the i(12p) chromosome, and clinical stage. No obvious differences were found in chromosomal constitution of metastatic and non-metastatic tumors. The results of the present study suggest that in TNSGCTs differences in clinical behavior are not associated with gross chromosomal differences. PMID- 8221607 TI - Childhood myelodysplastic syndrome with clonal evolution progressing to acute megakaryoblastic leukemia (ANLL-M7). AB - We treated a 16-month-old girl with myelodysplastic syndrome (MDS; refractory anemia with excess of blasts subtype, RAEB by FAB classification) that developed into acute megakaryoblastic leukemia (ANLL-M7). The blast cells were positive for CD41 shown by flow cytometry and for platelet peroxidase by electron microscopy. Cytogenetically, five kinds of abnormal karyotypes were apparent at the initial visit and karyotypic progression (clonal evolution) was also evident. These karyotypes were considered to be derived from the putative original clone, 48,XX, +6, +21. The observed karyotypes were considered 50,XX, +4,add(4)(q31), +6,add(7)(p22),add(10)(q24),add(12)(q11), +20, +21, + mar[karyotype A];48,XX,add(4)(q31), +6,add(10)(q24),add(12)(q11), +21 [karyotype B];48,XX, +6,t(6;13)(p23;q14), +21 [karyotype C];51,XX, +X, t(6;13)(p23;q14), + der(6)t(6;13)(p23;q14), +21, +21, + mar [karyotype D]; and 49,XX, +X, 3,t(6;13)(p23;q14), +der(6)t(6;13)(p23;q14), -12, +21, +21, + mar [karyotype E]. It seems karyotypes B and C were derived from the putative clone; karyotype B developed into karyotype A; and karyotype C developed into karyotype E through karyotype D. After development of ANLL-M7, the cytogenetic study showed a karyotype with further karyotypic progression. The patient was treated with high dose cytosine arabinoside (HD AraC) followed by allogeneic bone marrow transplantation. Despite intensive care, she died 3 months after the transplantation. PMID- 8221608 TI - In situ hybridization: a simple and sensitive method for detection of trisomy 12 in chronic lymphocytic leukemia. AB - Chromosome aberrations are detected in only 50% of patients with chronic lymphocytic leukemia (CLL), owing usually to the low mitotic rate exhibited by the neoplastic lymphocytes. Fluorescence in situ hybridization (FISH) is a simple method for identifying numerical abnormalities of the target chromosome in interphase nuclei. Therefore, we used the FISH procedure with chromosome 12 specific a-satellite probe to evaluate 19 patients with CLL. Trisomy 12 was detected in interphase cells of 12 patients (63%). Cytogenetic analysis, performed in nine patients, yielded trisomy 12 in four (44%). FISH detected three patients with trisomy 12 in whom conventional cytogenetic method yielded a normal karyotype. FISH is a simple, reliable, and sensitive method for detection of trisomy 12 in patients with CLL. PMID- 8221609 TI - X-ray-induced chromatid damage in cells from Down syndrome and Alzheimer disease patients in relation to DNA repair and cancer proneness. AB - Frequencies of chromatid aberrations in response to G2-phase x-irradiation were compared in PHA-stimulated blood lymphocytes from healthy control subjects, Down syndrome (DS) patients, and Alzheimer disease (AD) patients. In cells arrested with Colcemid immediately (0-30 min) after x-irradiation, DS, AD, and control cells showed similar high frequencies of chromatid breaks and gaps, representing unrepaired DNA strand breaks. Frequencies had decreased in AD and control cells arrested 30-90 min after irradiation. However, DS cells had two- to three-fold higher frequencies than AD or control cells. This result indicates deficient repair of the DNA damage in DS cells. Similar responses were obtained with lymphocytes from four of seven DS parents tested and with skin fibroblasts from DS patients compared to age-matched controls. Addition of 1-beta-D arabinofuranosylcytosine (ara-C), an inhibitor of the repair polymerase, after x irradiation during G2 phase increased the frequencies of chromatid breaks and gaps in lymphocytes from control and AD donors significantly more than in those from DS patients. This result indicates a deficiency in DS cells in incision at sites of x-ray-induced damage. Thus DS, like other cancer-prone genetic disorders, has a G2-phase DNA repair deficiency in strand break repair and also a second DNA repair deficiency in incision activity. PMID- 8221610 TI - Sequence of centromere separation. Minor satellite DNA does not influence separation of inactive centromeres in transformed cells of mouse. AB - Neoplastic cells may carry inactive centromeres on some multicentric, yet stable, chromosomes. We report that some inactive centromeres in L929 mouse cells do not contain minor satellite DNA, the DNA fraction which has been suggested to constitute the centromere. We compared the sequence of separation of inactive centromeres carrying the minor satellite with those lacking this fraction. The sequence of separation appears to be independent of whether or not the inactive centromeres carry the minor satellite DNA. The timing of replication of the inactive centromeres is also independent of this DNA. Hence, minor satellite of mouse is not a factor in holding together the subunits of inactive centromeres. Extension of these results to active centromeres might suggest that the minor satellite DNA is not a factor responsible for adhesion of the two centromere sub units up until late meta-anaphase. PMID- 8221611 TI - Influence of M-BCR breakpoint sites on the duration of chronic phase in 100 patients with chronic myelocytic leukemia. AB - Rearrangements of the bcr (M-BCR) gene were studied in 100 patients with chronic myelocytic leukemia (CML). To determine the significance of a chimeric gene expression in the progression of CML, we analyzed 43 patients for bcr-ABL chimeric mRNA expression. Both DNA and RNA analyses revealed a possible influence of breakpoint sites in the bcr region on the duration of the chronic phase. Patients with the breakpoint located at about the 1-kb region between BamHI and HindIII in bcr exon 3 (region C2) had a significantly shorter chronic phase (31 months) (p = 0.028) than patients in whom the breakpoint was located in other regions. When the bcr locus was divided into 5' and 3' regions as for the BamHI cleavage site located near the 5' region of bcr exon 3, the chronic phase duration in patients with the 5' site (HindIII-BamHI) and 3' site (BamHI-EcoRI site) was 75 and 38 months, respectively. However, the difference was not statistically significant (p = 0.128). These results suggest that only the breakpoint site at C2 on the bcr locus, rather than breakpoint sites in other regions, has an important role in the progression of CML. PMID- 8221612 TI - Loss of tumorigenicity in a methotrexate-resistant human leukemia cell line. AB - We have studied the tumorigenicity of a CCRF-CEM-derived cell line (CEM/MTX-3) resistant to MTX. Eight of nine mice inoculated with drug-sensitive CEM cells developed tumors within 5 weeks, but 16 weeks after inoculation with CEM/MTX-3 cells, none of nine mice developed tumors. We were unable to detect dihydrofolate reductase gene overexpression, amplification, or rearrangement in CEM/MTX-3 cells. Instead, the resistance in CEM/MTX-3 cells appeared to be due largely to decreased methotrexate accumulation. Because tumorigenicity could have been related to intracellular folate levels, we cultured CEM and CEM/MTX-3 cells in folate-rich and folate-deprived media. When inoculated in mice, CEM cells cultured in either medium rapidly formed tumors. As before, CEM/MTX-3 cells grown in either medium did not, suggesting that factors other than low folate levels contributed to the inability of CEM/MTX-3 cells to form tumors. Cytogenetic analysis revealed that the CEM/MTX-3 karyotype contained a unique and complex translocation marker chromosome that was not observed in the CEM cell line and which involved chromosomal breakpoints at bands 11p14, 22p11, and 22p13. Although biochemical mechanisms are not yet delineated, this remodeled chromosome could be related to the loss of tumorigenicity in CEM/MTX-3 cells. PMID- 8221613 TI - Heterogeneity in bladder cancer as detected by conventional chromosome analysis and interphase cytogenetics. AB - Thirty transitional cell carcinomas (TCCs) of the bladder were examined by classical chromosome counting to establish range, modal number, and percentage of metaphases with 2n, 3n, 4n, and > or = 5n chromosomes. In addition, fluorescence in situ hybridization (FISH) was applied to interphase nuclei to detect the percentage of tumor cells showing polyploidization and chromosome imbalance. In FISH, centromere-specific DNA probes for chromosomes 1, 7, 9, and 11 were used. The tumors were analyzed flow cytometrically to determine the DNA index (DI). Fourteen of 21 cases (67%) having a DI = 1 showed, after classical chromosome counting, in addition to a diploid model number, some cells with a 3n and 4n chromosome count. With FISH, eight cases (38%) showed a low percentage of cells with multiple signals for each of the probes, thus indicating polyploidization. In 13 (62%) cases, an imbalance between different chromosomes was detected. In nine tumors having a DI of 1.6 to 1.9, classical chromosome counting showed low percentages of > or = 5n cells in four cases, in addition to a triploid modal number. With FISH in six cases, a low percentage of cells showed five or more signals for each of the chromosomes, indicating polyploidization. In all cases, a chromosome imbalance was detected. With classical chromosome counting not all tumors can be analyzed. With FISH, small percentages of polyploid cells are not recognized. Both methods complement each other in that chromosome counting allows readier detection of heterogeneity in DNA-diploid tumors after polyploidization, whereas FISH allows efficient recognition of the chromosomes involved in the process of imbalance. PMID- 8221614 TI - Age- and gender-related heterogeneity of cancer chromosome aberrations. AB - The karyotype of a neoplasm is known to be associated not only with the histopathologic subtype of the tumor but also with previous cytotoxic exposure and with the geographic place of origin of the patient. Some data also indicate that cytogenetic patterns vary with age and gender. To further investigate whether the frequencies of cancer chromosome aberrations differ between children and adults or between men and women, clinical and karyologic data on 14,141 neoplasms with clonal chromosome changes reported in the literature were assessed. In cytogenetically well-characterized neoplasias, recognized primary and secondary chromosome aberrations were selected, and their frequencies were calculated in men, women, children (< or = 15 years), and adults (> 15 years). In general, the frequencies of the various aberrations did not differ between men and women or between children and adults, but a few exceptions were found. In refractory anemia (RA) and RA with excess of blasts or in transformation, del(5q) was more common among women. In acute lymphoblastic leukemia (ALL-L1 + L2), t(1;19) was more frequently detected in women and del(6q) more common among men. In Philadelphia chromosome positive chronic myeloid leukemia, gain of an extra der(22)t(9;22) occurred more frequently among men. Four primary aberrations were more common in children than in adults: t(8;21) in acute myeloid leukemia (AML M2), -7 in AML-M4, der(11q) in AML-M5, and t(8;14) in ALL-L3. On the other hand, der(16q) in AML-M4 and t(9;22) in ALL-L1 + L2 were more common in adults. The only secondary cancer chromosome aberration showing a variation with age was loss of the Y chromosome in AML-M2 with t(8;21), being more common in children than in adults. These variations might be spurious and level out when more data are collected, but more probably they reflect, for reasons presently unknown, that different genetic mechanisms may be operative in children and adults--and even in men and women--in the development of some tumors. PMID- 8221615 TI - A unique EBV-negative low-grade lymphoma line (WSU-FSCCL) exhibiting both t(14;18) and t(8;11). AB - A new human B-cell line, WSU-FSCCL, was established from the peripheral blood of a patient with low-grade follicular small cleaved cell lymphoma in leukemic phase. Both the fresh lymphoma cells and the established cell line exhibit t(14;18)(q32;q21) and t(8;11)(q24;q21) chromosomal translocations, 6q-, 1p+, and +i(1q). PCR analysis confirmed the juxtaposition of the major breakpoint-cluster region of bcl-2 with immunoglobulin heavy chain (JH) gene rearrangements. Southern analysis demonstrated that the 8q24 breakpoint was 5' of c-myc exon 1. The new line grows as a single-cell suspension with a doubling time of approximately 26 hours. It expresses cytoplasmic and cell surface IgM-kappa and reacts with monoclonal antibodies to B-cell antigens. Cells are negative for T cell and myeloid/monocyte antigens as well as for Epstein-Barr virus nuclear antigen (EBNA). DNA histogram generated by flow cytometry indicated a near diploid stemline. While t(14;18) is common in follicular lymphomas, the t(8;11) is unusual in lymphomas, although it does involve a region frequently aberrant on chromosome 8. The rearrangement of c-myc may have conferred an aggressive clinical behavior seen in the terminal phase of the disease. The role of 11q21 remains undetermined. PMID- 8221616 TI - T-cell malignant lymphoma with a complex unbalanced translocation (8;11;14). PMID- 8221617 TI - Case of meningioma with del(1)(p32) as sole anomaly. AB - We studied a case of typical syncytial meningioma. Cytogenetic analysis of the tumor cells showed a karyotype with normal chromosomes 22 and only one anomaly, del(1)(p32). Cases of meningiomas with normal chromosomes 22 and other anomalies are rare, and it is difficult to correlate their histologic characteristics and biologic behavior. PMID- 8221618 TI - Deletion (X)(p11): another case of renal adenocarcinoma with involvement of Xp11. PMID- 8221619 TI - Acute myelomonocytic leukemia (M4) and t(15;17)(q24;q21). A diagnostic dilemma. PMID- 8221620 TI - An oncogenic cast to the chromosomes. PMID- 8221621 TI - The role of reduced growth rate in the development of drug resistance of HOB1 lymphoma cells to vincristine. AB - An MDR cell line resistant to 1.0 microM vincristine (designated HOB1/VCR1.0) was established. The growth rate of HOB1/VCR1.0 cells was slow. The cells did not go into active proliferation although the drug treatment was released for two months (revertant cells). The HOB1/VCR1.0 cell line and its revertant were resistant to a high dose of vincristine (up to 20 microM). These two cell lines showed a decrease in expression of the hyperphosphorylated form of the retinoblastoma protein, of which the hypophosphorylated form has been considered to be a tumor suppressor. Similar phenomena were observed in the parental cells surviving brief treatment with a lethal dose of vincristine (0.05 microM). The current study gives the impression that self-inhibited growth rate may participate in the initial drug resistance before the expression of P-glycoprotein when tumor cells are suddenly exposed to chemotherapeutic agents. PMID- 8221622 TI - Evidence that membrane proteins of rhabdomyosarcoma cell line RD bind human immunodeficiency virus type 1 (HIV-1). AB - Membrane proteins (MP) obtained from the human mesenchymal rhabdomyosarcoma cell line RD were coated on 96-well polystyrene microplates and tested for their ability to bind human immunodeficiency virus type 1 (HIV-1). The virus bound to MP was detected by solid phase assay. Anti-human CD4 monoclonal antibodies directed against the HIV-1 gp120 binding site of the CD4 receptor did not inhibit viral binding to MP. HIV-1 specific polypeptides were recovered from coated MP to microplates by a modification of the solid phase immunoisolation technique and shown by immunoblotting analysis using a high titer of biotinylated human anti HIV-1 IgG. Together these findings provide evidence that HIV-1 binding to RD cell surfaces can proceed via a mechanism other than those mediated by the CD4 receptor. PMID- 8221623 TI - Effect of alkyl sulfides on diazomethane-induced methylation of DNA in vitro. AB - Organosulfur compounds found in Allium species such as garlic and onions inhibit carcinogenesis induced by alkylating agents. One potential mechanism for this activity involves scavenging of ultimate carcinogenic species by the nucleophilic sulfur atom. Carcinogenic methylating agents such as methyl-nitrosourea produce, among others, O6-methylguanine and N7-methylguanine in DNA both in vivo and in vitro. In the present study we have determined the effect of several alkyl sulfides on the formation of O6-methylguanine and N7-methylguanine in DNA reacted with the methylating agent diazomethane in vitro. Dipropyl sulfide and diallyl sulfide affect guanine methylation by increasing the O6/N7 ratio without drastic alterations in the total amount of adduct formed. Three similar compounds- diallyl disulfide, allyl methyl sulfide and diallyl ether--had no appreciable effect on the amount of alkylation at either position. These data suggest that scavenging of diazomethane-like ultimate carcinogens does not play a major role in the inhibition of carcinogenesis by organic sulfides. PMID- 8221624 TI - Comparative cytotoxicities of selected minor dietary non-nutrients with chemopreventive properties. AB - The comparative acute cytotoxicities were determined for a varied spectrum of minor dietary non-nutrients that have been implicated as chemopreventive agents. Cytotoxicity was determined with the neutral red (NR) assay, using BALB/c mouse 3T3 fibroblasts as the bioindicators. Based on midpoint cytotoxicity (NR50) values, the range of cytotoxicity for the different chemicals varied by 1000 times. The sequence of potency was tannic acid, tamoxifen citrate, quercetin, benzyl and phenethyl isothiocyanate > glycyrrhetinic acid > indole-3-carbinol > caffeic acid > phytic acid > vanillin > ellagic acid > D-saccharic acid 1,4 lactone. Vanillin, at slight to moderately toxic concentrations, was the only test agent that induced multinucleation in the 3T3 fibroblasts. PMID- 8221625 TI - Quantitative analysis of the metabolism of 9,10-dihydrobenzo[a]pyrene by induced rat liver microsomes. AB - The ability of reduced polycyclic aromatic hydrocarbons to be converted to their fully aromatic forms by the microsomal cytochrome P-450 mixed-function oxidases may assist in the explanation of the mutagenic and tumorigenic activities of these agents. The metabolic conversion of 9,10-dihydrobenzo[a]pyrene (9,10 DHB[a]P) to benzo[a]pyrene (B[a]P) and 9- and/or 10-hydroxy-9,10-DHB[a]P (OH-9,10 DHB[a]P) was quantitatively measured. In beta-naphthoflavone-induced rat liver microsomes, 9,10-DHB[a]P was metabolized to B[a]P with a specific activity of 1.51 nmol B[a]P formed/min/mg microsomal protein. The formation of B[a]P was directly related to incubation time and microsomal protein concentration. Similarly, 9,10-DHB[a]P was converted to OH-9,10-DHB[a]P with a specific activity of 4.48 nmol OH-9,10-DHB[a]P formed/min/mg microsomal protein. Its formation was directly related to incubation time and microsomal protein concentration. The possibility of OH-9,10-DHB[a]P as a metabolic intermediate to B[a]P is discussed. PMID- 8221626 TI - Alternative splicing of the p53 tumor suppressor gene in the Molt-4 T lymphoblastic leukemia cell line. AB - The expression of the p53 tumor suppressor gene in ten human cell lines (nine cancers and one normal) was studied using reverse transcription, polymerase chain reaction (PCR) and direct sequencing. Using P53U and P53D primers for amplifying a 371-base pair (bp) target fragment spanning exons 7-10 of p53 cDNA, normal sized PCR products were amplified from 9 cell lines but not from the Hep3B hepatocellular carcinoma (HCC) cell line. An additional larger band (504 bp) was observed for the Molt-4 T-lymphoblastic leukemia cell line. Employing P531 and P53D primers which flank a 76-bp p53 cDNA fragment, 76 bp as well as 209 bp products were generated by PCR of Molt-4 cDNA. Direct sequencing of the 504 bp and 209 bp bands confirmed the presence of a 133 bp insertion between exons 9 and 10 in the aberrant transcript. This insertion was homologous to a 130-bp sequence within the wild-type p53 intron 9, except for 2 point mutations and 3 base insertions. Sequencing of P53U/P53D PCR products of Molt-4 genomic DNA revealed an 8 bp deletion just downstream to the 133 bp insertion, creating a novel donor splicing site within intron 9. This site, coupled with an inherent acceptor splicing site just upstream to the 133 bp insertion, suggests that the 133 bp stretch represents an alternative exon. The occurrence of a termination signal within this alternative transcript is predicted to culminate in a truncated p53 translational product. The sequences of the 371 bp PCR products of Molt-4, HT 1080, SiHa, CaSki, HeLa and MRC-5 cell lines corresponded with the wild-type p53 cDNA. G-->T transversions at the third base of codon 249 of p53 were detected in Mahlavu and PLC/PRF/5 HCC lines, while a TAC to CAC mutation at codon 234 was observed in an allele of the Raji Burkitt lymphoma line. PMID- 8221627 TI - Ribonucleotide reductase: a possible target for orotic acid induced mitoinhibition in normal hepatocytes in primary culture. AB - The present study was designed to determine the mechanism by which orotic acid, a rat liver tumor promoter, inhibits DNA synthesis in normal hepatocytes in primary culture. Our results indicate that orotic acid inhibited the epidermal growth factor induced expression (mRNA) of both M1 and M2 subunits of ribonucleotide reductase while the expression of c-fos, c-myc, c-Ha-ras and beta-actin was not inhibited to any significant extent. These studies suggest that ribonucleotide reductase may be one target for orotic acid-induced mitoinhibition. PMID- 8221628 TI - Effects of alprazolam on cellular immune response to surgical stress in mice. AB - Mice exposed to surgical stress induced by laparotomy and treated with chronic alprazolam (0.5-2 mg/kg) showed a dose-dependent reduction in stress-induced suppression of the natural killer (NK) cell activity. These immunoenhancing effects of alprazolam were more intense when it administered before the surgery was performed. PMID- 8221629 TI - Protective effects of green tea polyphenols administered by oral intubation against chemical carcinogen-induced forestomach and pulmonary neoplasia in A/J mice. AB - Our studies and others have shown the cancer chemopreventive effects of chronic administration of green tea in several animal tumor models. In this study, the administration of a polyphenolic fraction isolated from green tea (GTP) by oral intubation at a dose of 5 mg in 0.2 ml water 30 min prior to challenge with carcinogen, afforded significant protection against both diethylnitrosamine (DEN) and benzo(a)pyrene (BP)-induced forestomach and lung tumorigenesis in A/J mice. The protective effects were evident by a decrease in numbers of tumors/mouse in GTP-fed groups compared to non GTP-fed controls. In the forestomach tumorigenesis protocol, GTP afforded 71 and 66% protection against, respectively DEN- and BP induced tumor multiplicity. In the case of lung tumorigenesis protocol, however, the protective effects of GTP were 41 and 39%, respectively. Histological examination of forestomach tumors showed significantly lesser number of squamous cell carcinoma formation in GTP-fed groups of mice compared to carcinogen alone treated controls. When pulmonary tumors were examined histologically, no adenocarcinomas were observed in GTP-fed groups compared to 15% mice with adenocarcinomas in DEN and BP alone-treated controls. The results of this study suggest that limited doses of GTP administration by gavage 30 min prior to carcinogen challenge may afford protection against carcinogen-induced tumorigenesis in internal body organs. PMID- 8221630 TI - Suppressing effect of Lactobacillus casei administration on the urinary mutagenicity arising from ingestion of fried ground beef in the human. AB - It is known that the ingestion of cooked meat which contains carcinogenic heterocyclic amines causes increase in urinary mutagenicity in humans. Using 6 healthy non-smokers, we examined the effect of 3-week oral administration of Lactobacillus casei (bacilli commonly present in yoghurt), on the urinary mutagenicity derived from ingestion of fried ground beef. Comparison of the urinary mutagenicity found before and after the L. casei treatment showed that the treatment resulted in a decrease (6-67%, average 47.5%) of the mutagenicity. This suppressing effect is possibly related to the changes in the intestinal microflora population. PMID- 8221631 TI - Demonstration of selective protein complexes of p53 with 73 kDa heat shock cognate protein, but not with 72 kDa heat shock protein in human tumor cells. AB - It has been demonstrated that p53, especially, mutant p53 (mp53), makes protein complexes with major heat shock proteins hsp72/hsc73. However, there is no direct evidence showing whether hsp72 or hsc73 could bind preferentially to p53. In the present study, using TYKnu human ovarial carcinoma cells and monoclonal antibodies reacting specifically to hsp72/hsc73, we were able to find the selective protein complex formation with p53, presumably mp53, and hsc73, but not in the case of p53 and hsp72. The p53-hsc73 protein complexes dissociate with the addition of ATP, indicating that the dissociation is dependent upon the ATP hydrolysis. These data suggest that hsc73 rather than hsp72 plays an important role in the yet undefined mechanism of disregulated cell growth control by mp53. PMID- 8221632 TI - Mouse skin tumor-initiating activity of benz[j]aceanthrylene in SENCAR mice. AB - Benz[j]aceanthrylene (B[j]A), a cyclopenta-fused derivative of benz[a]anthracene, has been reported to be an active bacterial cell and mammalian cell gene mutagen, a morphological transforming agent in C3H10T1/2CL8 mouse embryo fibroblasts and a mouse lung tumorigen in strain A/J mice. B[j]A was evaluated as a skin tumor initiator in female SENCAR mice and was found to induce papilloma formation in the range of 40-400 micrograms/mouse. B[j]A was found to be extremely active, inducing 8.7 papillomas/mouse after an initiating dose of 40 micrograms/mouse. At this dose, 100% of the mice bore tumors. Comparison with four other cyclopenta fused polycyclic aromatic hydrocarbons suggests that B[j]A is extremely potent. PMID- 8221633 TI - Molecular modelling of tumour-promoting and non-promoting diterpene esters and molecular co-ordinates of the potent irritant resiniferatoxin. AB - The lowest energy conformer of seventeen diterpenes, representing five different phorbol and daphnane diterpene nuclei, has been generated. TPA possessed the highest minimum free energy of these compounds; all other compounds possessed a lower minimum free energy. Compounds based on the resiniferonol nucleus possessed the lowest minimum free energy (9.4-16.6% of that of TPA). The molecular co ordinates of the non-promoting but potent irritant resiniferatoxin (Rx) are also reported. These studies may be important in elucidation of the biochemical mechanisms of action of diterpene esters, including an understanding of the interactions of diterpene esters with the phorbol ester binding domain of the protein kinase C isoform family. PMID- 8221634 TI - Effects of media conditioned by a non-metastasizing human salivary gland adenocarcinoma cell clone and metastasizing clones from salivary gland and various other tissues on the proliferation, migration and protease production of bovine aortic endothelial cells in vitro. AB - We demonstrate the role in tumor-associated angiogenesis of factors released into conditioned medium (CM) from in vitro human salivary gland cell clones with biological phenotypes ranging from non-metastasizing to metastasizing. A non metastasizing human salivary gland adenocarcinoma cell clone HSGc and its subclone with metastatic potential (Gc2-100 cl-1) were employed. We also used metastasizing cell clones obtained by explant cultures of organs of Gc2-100 cl-1 tumor-bearing nude mouse. The proliferation and migration of bovine aortic endothelial (BAE) cells were significantly stimulated by the addition of CM obtained from Gc2-100 cl-1 and metastasizing cell clones, while CM from HSGc was ineffective. When the effect on protease secretion by BAE cells was examined, CM from Gc2-100 cl-1 and metastasizing cell clones inhibited the secretion of type IV collagenases by BAE cells much more than did CM from HSGc. These findings, therefore, may imply that Gc2-100 cl-1 and metastasizing cell clones secrete angiogenic factors that stimulate not only the proliferation and migration of endothelial cells but also the formation of basement membrane components necessary for the reconstruction of new blood vessels at migrated sites of endothelial cells by preventing the degradation of basement membrane component, type IV collagen. PMID- 8221635 TI - Studies on the mutagenic activation of heterocyclic amines by cynomolgus monkey, rat and human microsomes show that cynomolgus monkeys have a low capacity to N oxidize the quinoxaline-type heterocyclic amines. AB - A number of mutagens and carcinogens have been isolated from cooked meats. In the current study we investigated the ability of hepatic microsomes from cynomolgus monkeys, Fischer-344 rats and humans to metabolically activate these compounds. Monkeys had almost no capacity to activate the quinoxaline-type compounds to mutagens in the Ames test relative to rats and humans but were able to activate the quinoline, pyridoindole and pyridoimidazole compounds. Differences in the mutagenicity of the quinoline and quinoxaline compounds by monkeys and rats was related to differences in cytochrome P-450-mediated N-oxidation between the species. This suggests that monkeys and rats may have different hepatic cytochrome P-450 isozymes, which are important for the metabolic activation of quinolines and quinoxalines, or that the orthologous monkey cytochromes show a select substrate specificity for the quinolines over the quinoxalines. PMID- 8221636 TI - Protective and curative potential of vaccination with interleukin-2-gene transfected cells from a spontaneous mouse mammary adenocarcinoma. AB - The potential of interleukin 2-gene-transfected tumor cells to prevent tumor growth and cure established tumors was evaluated using cells from a spontaneous, invasive, and metastasizing mouse mammary adenocarcinoma. Tumor cells engineered to secrete interleukin 2 initially trigger a local inflammatory reaction that leads to inhibition of established parental adenocarcinomas, as well as an antigenically unrelated fibrosarcoma. The ensuing systemic immunity selectively inhibits subsequent parental cell challenges and cures established parental adenocarcinomas and their lung metastases, although less effectively as the neoplastic mass increases. Multiple injections of interleukin 2-gene-transfected tumor cells may thus be considered a new form of vaccination in the management of minimal residual disease and incipient metastases. PMID- 8221637 TI - Polymerase chain reaction analysis of parathyroid hormone-related protein gene expression in breast cancer patients and occurrence of bone metastases. AB - Parathyroid hormone-related protein (PTHrP) is associated with the syndrome of humoral hypercalcemia of malignancy. A high incidence of positive staining for PTHrP is observed in breast cancer and positivity is more frequent in patients who develop bone metastases. We assessed the presence of PTHrP mRNA by using the polymerase chain reaction in 38 normocalcemic breast cancer patients with long term follow-up (minimum, 5 years) selected for the presence or absence of later bone metastasis development. In all the patients except one, the PTHrP gene was expressed in the breast tumor. The level of amplified PTHrP complementary DNA was inversely related to age (P < 0.02) and positively related to the proportion of invaded nodes (P < 0.02) but was not related to the other usual prognostic factors. The level of PTHrP mRNA was not different between the group of patients without recurrence or metastases (n = 11) and the group of patients who later developed metastases in soft tissues (n = 10). By contrast, patients who subsequently developed bone metastases (n = 17) showed higher PTHrP gene expression than patients in the other two groups (P < 0.001). This study suggests that strong PTHrP gene expression in breast tumors is associated with the onset of bone metastases. PMID- 8221638 TI - Coexistence of somatic and germ-line mutations of APC gene in desmoid tumors from patients with familial adenomatous polyposis. AB - Desmoid tumors, which are locally invasive with recurrence but without metastasis, are frequently observed in patients with familial adenomatous polyposis after abdominal surgery or during pregnancy. This study analyzed mutation of the adenomatous polyposis coli gene in 8 desmoid tumors from 7 familial adenomatous polyposis patients using polymerase chain reaction-single strand conformation polymorphism and the direct sequencing method. Seven somatic mutations, 1 somatic allele loss, and 6 germ-line mutations were detected. The majority of adenomatous polyposis coli gene mutations were deletions of 1 to 19 base pairs in exon 15, and all mutations led to the formation of stop codons. A somatic mutation with repetition of 82 base pairs from codon 1399 to 1426 was also observed in a desmoid, which was most likely caused by an error during replication or repair replication. No mutation was detected in exons 1 to 2 of H ras, K-ras, and N-ras genes and in exons 5 to 8 of p53 gene, in these tumors. The simultaneous existence of somatic and germ-line alterations of adenomatous polyposis coli gene observed in all 8 tumors strongly suggests that inactivation of both alleles of adenomatous polyposis coli gene is involved in the development of desmoid tumors. PMID- 8221640 TI - Genetic instability in pancreatic cancer and poorly differentiated type of gastric cancer. AB - To examine genetic instability during carcinogenesis, we screened 171 carcinomas of the breast, liver, proximal colon, stomach, pancreas, uterine cervix, and ovary for replication error at four microsatellite marker loci on chromosome 2, 3, and 17. A significantly high incidence of genetic instability was observed in pancreatic (6 of 9 tumors) and gastric cancers (22 of 57 cases). In other types of carcinoma, the incidence of replication error-positive cases was relatively low (3-16%). Among gastric carcinomas, significantly more replication error positive cases were observed in the poorly differentiated types (16 of 25 cases) than in well differentiated types (3 of 18) (P = 0.0023 by Fisher's exact test). Our results suggested that genetic instability is likely to play an important role in development of pancreatic and gastric cancers, particularly poorly differentiated adenocarcinomas. PMID- 8221639 TI - Linkage analysis of DRD2, a marker linked to the ataxia-telangiectasia gene, in 64 families with premenopausal bilateral breast cancer. AB - Recent reports suggest that subjects who are heterozygous for the ataxia telangiectasia gene are at increased risk of breast cancer. We conducted linkage analyses of 64 families with premenopausal bilateral breast cancer using DRD2, a marker linked to the ataxia-telangiectasia locus at 11q22-23. We assumed a model with dominant transmission of breast cancer. Lod scores summed over all families provided strong evidence against tight linkage (e.g., a lod score of -6.08 at theta = 0.00001), although a single family provides suggestive evidence of tight linkage to DRD2. Evidence against linkage to 11q was strongest among families that may involve the BRCA1 breast cancer susceptibility gene on 17q21. However, we did not observe evidence of linkage to 11q among the remaining subgroup with neither a family history of ovarian cancer nor the appearance of linkage to 17q21. PMID- 8221641 TI - High affinity binding sites for gastrin releasing peptide on human gastric cancer and Menetrier's mucosa. AB - The bombesin-like peptides gastrin releasing peptide (GRP) and neuromedin B are found in the submucosal and myenteric plexuses of the human gastrointestinal tract. These peptides are potent mitogens to Swiss 3T3 fibroblasts and are important autocrine growth factors in human small cell lung cancer cells. We have recently described the presence of receptors for the bombesin-like peptide, GRP, on the human gastric cancer cell line St42. In this study, we examined fresh resected gastric cancer and uninvolved mucosa from 23 patients for the presence of binding sites to the bombesin-like peptides. Thirteen of 23 gastric cancers expressed high affinity binding sites for bombesin (mean Kd = 3.42 nM; mean Bmax = 40.5 pmol/mg protein), of which 12 were subsequently characterized and found to be of the GRP-preferring subtype. One of 23 mucosal samples specifically bound bombesin and was the only sample from a patient with Menetrier's disease, a disorder of mucosal growth known to be premalignant. The early gastric cancer from this patient also possessed high affinity binding sites for GRP. This is the first description of binding sites to bombesin-like peptides on human gastric cancer and Menetrier's mucosa. The role of bombesin/GRP antagonists in the treatment of gastric cancer warrants investigation. PMID- 8221642 TI - Evidence for two bladder cancer suppressor loci on human chromosome 9. AB - Most carcinomas of the bladder show loss of heterozygosity for markers on human chromosome 9, which suggests that one or more tumor suppressor genes are located on this chromosome. Several observations suggest that such alterations are an important early step in tumorigenesis. We analyzed the pattern of allelic loss in 46 primary carcinomas of the bladder using 19 polymorphic markers from chromosome 9. While most tumors with allelic loss showed loss of heterozygosity for all informative markers that were tested, six tumors demonstrated only partial loss of chromosome 9. Two tumors with partial loss contained deletions that predominantly involved the q arm, as shown by previous studies. The other four tumors contained deletions that predominantly or exclusively involved the p arm, with a common region of loss between D9S161 (9p21) and the telomere. The results show that there is no single common region of loss on chromosome 9 and identify two distinct regions of loss that may contain bladder tumor suppressor loci. PMID- 8221643 TI - Simultaneous analysis of cell cycle kinetics at two different DNA ploidy levels based on DNA content and cyclin B measurements. AB - The methods of cell cycle analysis that rely on DNA content measurements cannot discriminate between cells at different phases of the cycle if these cells have similar DNA content. This limitation can be circumvented by measurement of another cell cycle phase-specific cell constituent in addition to DNA content, followed by bivariate analysis of the correlated data. The aim of the present study was to explore the utility of a monoclonal antibody against the G2- and M phase-specific regulatory protein cyclin B for discrimination of cell populations with overlapping DNA content. This analysis, which was based on correlated DNA/cyclin B content measurements by flow cytometry, was applied to human lymphocytic leukemic MOLT-4 cells. The onset of cyclin B synthesis was observed in the last one third of S phase with its maximum accumulation in G2 and M phases; cells in G1 and early- and mid-S phases were negative. Cells arrested in metaphase by vinblastine expressed high levels of this protein, although not as high as in cells arrested in G2 by the DNA topoisomerase II inhibitor m-AMSA. Disruption of cytokinesis by the protein kinase inhibitor staurosporine led to DNA rereplication, cell progression through the chromatin cycle at higher DNA ploidy, and induction of polyploidy. It was possible, utilizing the cyclin B antibody, to discriminate between G2 + M cells with a 2C level of DNA and G1 cells with 4C DNA, as well as to distinguish doublets of G1 cells with a 2C DNA level. Thus, the rate of cell entrance to G1 at the 4C DNA level and the rates of progression through the cycle at both the 2C and 4C DNA levels could be simultaneously estimated. The data indicate that, in the presence of 0.1 microM staurosporine, cytokinesis of all MOLT-4 cells is impaired and the cells enter to and progress through the chromosome cycle at 4C DNA at the same rates as at 2C DNA. This approach can be helpful in the analysis of DNA ploidy and the cell cycle of human tumors when there is an overlap in DNA content values between normal stromal or infiltrating cells and aneuploid tumor cell population and may be the method of choice to investigate the activity of antitumor drugs which impair cytokinesis but do not interfere with progression of cells through the chromatin cycle. PMID- 8221644 TI - Genetic instability of microsatellites in endometrial carcinoma. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is characterized by a familial predisposition to colorectal carcinoma and extracolonic cancers of the gastrointestinal, urological, and female reproductive tracts, notably the endometrium. A genetic locus for HPNCC was recently determined by linkage analysis to exist on chromosome 2p; both sporadic and HNPCC-associated colorectal carcinomas exhibit a "replication error" phenotype, characterized by instability of dinucleotide repeat sequences throughout the genome. Here, we address the hypothesis that the replication error phenotype would be evident in some fraction of sporadic endometrial carcinomas or in those associated with HNPCC. Microsatellite instability was observed in 17% of sporadic endometrial carcinomas and in 75% of those tumors associated with HNPCC. These data indicate that the HNPCC gene is also involved in heritable and somatic forms of endometrial carcinoma. PMID- 8221645 TI - Elevated production of active oxygen in Bloom's syndrome cell lines. AB - Based on our previous evidence indicating that the elevated sister chromatid exchange that characterizes Bloom syndrome (BS) cells may arise in response to elevated production of active oxygen, we have quantitated the levels of active oxygen in two control, two BS and one BS revertant cell lines. Luminol-dependent chemiluminescence was used as a measure of active oxygen production following treatment of the cells with the calcium ionophore A23187 or the chemotactic tripeptide N-formylmethionylleucylphenylalanine. A peptide factor present in plasma was required for priming the cells to undergo the oxidative response. As determined with A23187, active oxygen production was elevated in BS cell lines by 48.6% above control. Using N-formylmethionylleucylphenylalanine, active oxygen production was found to be increased by 250-314%. Chemiluminescence was inhibited in a dose-dependent manner by diphenylene iodonium, which specifically binds to and inhibits membrane-associated NADPH oxidase activity. This compound inhibited oxygen radical production nearly 3 times more effectively in control cells than in BS cells. The capacity to produce elevated levels of oxygen radicals may contribute to the spontaneous chromosomal instability of BS cells and to the associated high incidence of neoplasia in individuals with BS. PMID- 8221646 TI - Fusion of PAX3 to a member of the forkhead family of transcription factors in human alveolar rhabdomyosarcoma. AB - Alveolar rhabdomyosarcoma, a malignant tumor of skeletal muscle, is characterized by a chromosomal translocation, t(2;13)(q35;q14). This translocation is associated with a structural rearrangement of the gene encoding PAX3, a presumed transcriptional regulator expressed exclusively during embryogenesis. The breakpoint results in a fusion between PAX3 and a gene provisionally named ALV, a novel member of the forkhead family of transcription factors. In PAX3-ALV, the structural integrity of both PAX3 DNA-binding regions, the paired box and homeodomain, are retained while the putative transcriptional activation domain of PAX3 is replaced by the bisected forkhead DNA-binding domain of ALV. Formation of chimeric transcription factors has now been implicated in diverse human tumors of myogenic, hematopoietic, neuroectodermal, and adipocytic origin, suggesting that transcriptional deregulation is a common mechanism of tumorigenesis. PMID- 8221647 TI - Recent advances in head and neck cancer--larynx preservation and cancer chemoprevention: the Seventeenth Annual Richard and Hinda Rosenthal Foundation Award Lecture. PMID- 8221648 TI - Involvement of human liver cytochrome P450 3A in vinblastine metabolism: drug interactions. AB - Vinblastine biotransformation was investigated by using a human liver microsomes library. The drug was converted into one major metabolite (M) upon incubation with the microsomes. A large interindividual variation in vinblastine metabolism was observed among the samples tested, with a 4.4 ratio between the lowest and the highest metabolic rates. The biotransformation of vinblastine followed Michaelis-Menten kinetics (Km = 6.82 +/- 0.27 microM and Vmax = 0.64 +/- 0.06 nmol/min/mg protein). The involvement of the cytochrome P450 3A subfamily in vinblastine metabolism was demonstrated by the following body of evidence: (a) the competitive inhibition of vinblastine biotransformation by cytochrome P450 3A specific probes with Ki values of 0.17, 22.5, 14.8, and 35.3 microM for ketoconazole, erythromycin, troleandomycin, and vindesine, respectively; (b) the immunoinhibition of vinblastine metabolism by polyclonal anti-cytochrome P450 3A antibodies; (c) the highly significant correlation between the level of cytochrome P450 3A determined by Western blots and vinblastine metabolism (r = 0.759, P < 0.001); (d) the highly significant correlation between erythromycin N demethylase activity (mediated by cytochrome P450 3A) and vinblastine metabolism (r = 0.83, P < 0.001); (e) the significant correlation between the CYP3A4 mRNA level and vinblastine metabolism (r = 0.60, P < 0.1). Although vincristine and navelbine (members of the Vinca alkaloid family) also inhibit the metabolism of vinblastine, suggesting the involvement of the cytochrome subfamily in their respective metabolisms, other anticancer drugs currently associated with vinblastine in chemotherapy (etoposide, Adriamycin, lomustine, and teniposide) also interfere with vinblastine biotransformation. These metabolic drug interactions may alter the antitumor activity and/or toxicity of the drug during anticancer chemotherapy. PMID- 8221649 TI - Cytochrome P-450-induced cytotoxicity of mitoxantrone by formation of electrophilic intermediates. AB - Recent studies of our group have shown that the oxidation of the substituted anthraquinone skeleton is involved in the biotransformation of mitoxantrone. In this report the importance of this process with regard to the mode of action of the drug is investigated. This communication describes a new high performance liquid chromatography separation for mitoxantrone and its metabolites allowing the direct coupling of high performance liquid chromatography to mass spectrometry. Application of this technique to bile of mitoxantrone-treated pigs reveals the formation of several metabolites in addition to the drug-derived compounds found in urine. Seven biliary metabolites are identified as thioether derivatives of mitoxantrone and its side chain oxidation products. Independent synthesis and structural elucidation of 3 thioether conjugates of the drug provides unequivocal evidence that the hydroquinone moiety of mitoxantrone is the site of reaction with glutathione. Furthermore, the formation of the thioether conjugates in HepG2 hepatoma cells and in rat hepatocytes during cell incubations is demonstrated. Inhibition of cytochrome P-450 with metyrapone prevents the formation of the thioether conjugates and leads to a complete loss of the cytotoxicity of mitoxantrone in HepG2 cells and rat hepatocytes up to concentrations of 200 to 400 microM thereby indicating that mitoxantrone has a negligible effect by itself. Rat hepatocytes were found to be more susceptible for the oxidation-induced cytotoxicity than HepG2 cells. These results demonstrate that the acute cytotoxicity of mitoxantrone depends on prior oxidation of its 1,4-dihydroxy-5,8-diaminoanthraquinone moiety. PMID- 8221650 TI - Purine nucleotide levels in host tissues of Ehrlich ascites tumor-bearing mice in different growth phases of the tumor. AB - The pool sizes of purine nucleotides, nucleosides, and nucleobases were measured in the host tissues liver, skeletal muscle, and blood of Ehrlich ascites tumor bearing mice during the different periods of tumor growth. There were large differences of tissue concentrations of these metabolites between control animals, animals in the logarithmic growth period of the Ehrlich ascites tumor, and animals in the resting phase of tumor growth. The ATP concentrations in liver, muscle, and erythrocytes were higher during the proliferating phase of the tumor compared with the ATP levels of these organs in healthy animals. In liver and skeletal muscle the ATP concentration decreased during the transition from proliferating into resting phase of tumor growth. The concentrations of nucleosides and nucleobases within the RBC and blood plasma deceased during the logarithmic growth phase but restored during the plateau period. As well as in the organs/cells investigated and in the body fluids (plasma, ascites fluid) a tremendous increase of adenosine concentration during the resting phase of tumor growth was observed. From changes of total purine ribonucleotide pattern an activation period in the nucleotide metabolic pathways of liver and skeletal muscle during the proliferating phase of tumor growth is postulated. PMID- 8221651 TI - Migratory pattern of fetal rat brain cells and human glioma cells in the adult rat brain. AB - The migratory behavior of two human glioma cell lines (D-54MG and GaMG) and fetal rat brain cells grafted into the adult rat brain was studied. To trace the implanted cells, they were stained with the carbocyanine vital dye 1,1' dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate before injecting them into the white matter above the corpus callosum. The animals were sacrificed 2 h and 7 and 21 days after injection, and the brains were removed and cryosectioned. Fluorescence microscopy showed that both the 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate-stained fetal and tumor cells had the same migratory pattern. Implanted cells were found along myelinated fibers in the corpus callosum and in the perivascular space. After immunostaining for several extracellular matrix (ECM) components (laminin, fibronectin, collagen type IV, and chondroitin sulfate), laminin deposits were observed in the border zone between the host tissue and implanted tumor cells as well as fetal cells. By using two different types of antibodies against fibronectin, it is shown that the fibronectin expression observed in the tumor matrix may be host derived. This was further supported by the fact that tumor spheroids obtained from the two glioma cell lines were negative when immunostained for these ECM components. Several of the ECM components may be host derived. This can be caused by neovascularization and repair synthesis or by a local production of guiding substrates which are important for tumor cell locomotion. The present data suggest that the migratory patterns of fetal and glioma cells are indistinguishable when transplanted into the adult rat brain. Thus, glioma cells may be routed by the same ECM components that play a major role during brain development. PMID- 8221652 TI - Insulin-like growth factor II and WT1 transcript localization in human fetal kidney and Wilms' tumor. AB - In situ hybridization was used to examine, in parallel, the localization of insulin-like growth factor II (Igf2) and WT1 transcripts in normal fetal kidney and Wilms' tumor. The expression of Igf2 and WT1 transcripts in the fetal kidney is almost complementary in both the epithelial and stromal cell lineages derived from the undifferentiated metanephrogenic blastema. The patterns of transcription of Igf2 in three Wilms' tumors appeared to be perturbed as compared to the normal fetal kidney. In these tumors Igf2 transcripts were detected in structures that are developmentally equivalent to the renal vesicle, which in the normal kidney do not contain Igf2 transcripts. These results suggest that Wilms' tumors arise from an alteration in the regulation of Igf2 mRNA synthesis. PMID- 8221653 TI - Enhancement of rat urinary bladder tumorigenesis by lipopolysaccharide-induced inflammation. AB - Chronic inflammation of the urinary tract is a significant risk factor for the development of urinary bladder cancer in humans. We previously demonstrated that weekly treatment with killed Escherichia coli enhanced rat urinary bladder tumorigenesis initiated by the carcinogen N-methyl-N-nitrosourea. We conducted the present study to determine whether lipopolysaccharide (LPS), a major cell wall component of E. coli, had a tumor-enhancing effect. LPS was instilled twice a week at three doses (100, 1.0, and 0.01 microgram/ml) into heterotopically transplanted rat urinary bladders which were treated with a single low dose (0.25 mg) of N-methyl-N-nitrosourea or vehicle. Rats treated with 100 micrograms/ml of LPS showed a significant increase in the incidence and number of tumors in the bladders pretreated with N-methyl-N-nitrosourea. Treatment with LPS alone did not induce tumors. The enhancing effects were associated with a marked increase in the numbers of polymorphonuclear leukocytes and an increase in the H2O2 concentration in the bladder lumen. Oxidative stress by reactive oxygen intermediates and a proliferative response of the carcinogen-exposed urothelium to the inflammatory stimulation appeared to play a significant role in tumor enhancement by LPS. PMID- 8221654 TI - Phase I trial of an oral immunomodulator and interferon inducer in cancer patients. AB - Imiquimod [1-(2-methylpropyl)-1H-imidazo[4,5c]quinolin-4-amine] is a compound of low molecular weight that, when administered p.o., induces interferon-alpha in several animal species and inhibits tumor growth in mice. To determine maximum tolerated dose, toxicity, and biological response in humans, a phase I clinical trial was conducted with 14 eligible cancer patients who received 100-500 mg imiquimod p.o. either once or twice weekly. Imiquimod induced interferon-alpha in serum in 10 of 19 doses of 200-300 mg. Interferon serum levels peaked 8-24 h after treatment and reached a maximum of 23,000 IU/ml in one patient. Significant mean increases (P < 0.01) in serum beta 2-microglobulin (1.5-fold), serum neopterin (3.5-fold), and 2-5A synthetase activity in peripheral blood mononuclear cells (7.9-fold) indicated that 200-300 mg imiquimod had biological and immunological activity in all evaluable patients. Increases in serum interferon, beta 2-microglobulin, and neopterin correlated significantly with dose (P < 0.001). No patient developed measurable antibody to interferon-alpha. Dose-limiting side effects included fatigue, malaise, fever, headache, and lymphocytopenia; no hepatic or renal toxicity or other hematological changes exceeded the normal range. Patients tolerated weekly doses of up to 500 mg, with the longest treatment lasting 4 weeks at 200 mg weekly. Twice-weekly doses up to to 300 mg were tolerated, with the longest twice-weekly treatments being 200 mg for 9 weeks and 100 mg for 25 weeks. No clinical responses were observed. Imiquimod, as an oral inducer of interferon, may have therapeutic usefulness in human cancers, viral infections, and other diseases. However, before initiation of phase II trials, additional work will be required to establish a tolerated dose and schedule for continued administration. PMID- 8221655 TI - Individualized chemotherapy for patients with non-small cell lung cancer determined by prospective identification of neuroendocrine markers and in vitro drug sensitivity testing. AB - We attempted to prospectively select individualized chemotherapy for 165 non small cell lung cancer patients based on in vitro analysis of neuroendocrine (NE) markers and drug sensitivity testing (DST) using fresh tumor. The chemotherapy used for small cell lung cancer (SCLC) was selected when NE marker expression determined by L-dopa decarboxylase assay was documented. Selection of chemotherapy for other patients was guided by DST results using a modified dye exclusion assay when available; otherwise etoposide and cisplatin was administered. A total of 112 of 165 (68%) specimens were assayed for L-dopa decarboxylase and 36 patients (22%) had DST. In vitro data directed management for 27 of 96 (28%) patients given chemotherapy: 6 with NE markers were treated with the SCLC regimen; and 21 (58% of those with DST) received their DST-selected chemotherapy regimen. There were no significant differences in response rate among all 3 treatment arms (P = 0.076). However, response to chemotherapy for the patients treated prospectively with a SCLC regimen was 3 of 6 (50%), marginally better than patients given their DST-selected chemotherapy regimen (2 of 21; 9%; P = 0.056) or those treated with etoposide and cisplatin (10 of 69; 14%; P = 0.061). When patients whose NE markers were identified retrospectively are included, 4 of 9 (44%) responded to administered chemotherapy, compared to 7 of 55 (13%) with no NE markers present (P = 0.04). There were no differences in survival among the three treatment groups. Cisplatin and etoposide comprised the most active regimen in vitro for tumors from 16 of 36 (44%) patients, potentially limiting the benefit of DST since this is often the empiric therapy for non-SCLC. Furthermore, the correlation between in vitro and clinical response is nonsignificant for all drugs tested, highlighting the overall relative resistance of non-SCLC tumors to currently available chemotherapy. PMID- 8221656 TI - Improved prediction of survival in advanced adenocarcinoma of the ovary by immunocytochemical analysis and the composition adjusted receptor level of the estrogen receptor. AB - Conventional cytosol estrogen receptor analysis is not a significant prognostic variable in serous ovarian carcinoma. Although the use of immunocytochemical receptor analysis for estrogen does provide prognostically useful information in enhanced accuracy of predicting survival in patients with ovarian cancer, its usefulness can still be improved. Surgical samples from ovarian carcinomas are heterogeneous in tissue composition. Immunocytochemical receptor analysis allows for the specific assessment of the tumorous portions of a histological specimen. However, it is limited by its dependence on staining intensity as the determining factor. Biochemical receptor analysis does provide objective information concerning the number of receptor molecules present in a given sample, but the value is not adjusted for histological composition of the tumor section. Therefore, we have attempted to combine the advantages of both methods. By adjusting the conventional receptor analysis for the percentage of tumor present in the specimen, we have eliminated the tissue heterogeneity as a confounding variable. The resulting value is named Composition Adjusted Receptor Level or CARL. A prospective study was performed on the estrogen receptor concentrations in 61 ovarian cancers. Minimum follow-up was 8 years. For the percentage of tumor in the specimen, a highly significant correlation of the assessment of the two pathologists was observed. Stage (P < 0.05) and grade (P < 0.05) as well as cell type (P < 0.05) were found to be significant prognostic variables. In an attempt to eliminate the confounding influences of these variables, the CARL of the estrogen receptor was assessed with regard to its prognostic significance in 32 grade 2 and 3 serous carcinomas of the ovary, stage III and IV. A linear correlation between CARL and survival was found above a threshold estrogen receptor concentration of 15 fmol/mg cytosol protein using a correlation of the Cox proportional hazards model (P < 0.02). Our data suggest that (a) the assessment of the percentage of tumor in a given sample is not significantly observer dependent, (b) CARL is a significant predictor of survival in serous ovarian carcinoma, and (c) a CARL should be determined for the analysis of any cytosol receptor in solid tumors. PMID- 8221657 TI - CCKA and CCKB receptors are expressed in small cell lung cancer lines and mediate Ca2+ mobilization and clonal growth. AB - Gastrin, cholecystokinin (CCK), and CCK-related peptides comprise a hormonal family characterized by an identical carboxy-terminal amino acid sequence, a domain critical for receptor binding. The addition of gastrin to small cell lung cancer (SCLC) cells causes a rapid and transient increase in the intracellular concentration of calcium ([Ca2+]i). Furthermore, gastrin acts as a direct growth factor through CCKB/gastrin receptors. We report here that the expression of the mRNA coding for CCKB/gastrin receptors correlates with the responsiveness of SCLC cells to gastrin in terms of Ca2+ mobilization and stimulation of clonal growth in semisolid medium. The GLC19 SCLC cell line had no detectable expression of CCKB/gastrin receptor mRNA. Accordingly, gastrin (1-100 nM) did not cause any measurable increase in [Ca2+]i. In contrast, the addition of cholecystokinin residues 26-33 (CCK-8) caused a rapid and transient increase in [Ca2+]i in this cell line. CCK-8 mobilized Ca2+ in a dose-dependent manner in the nanomolar range (half-maximal stimulatory concentration = 12 nM). Furthermore, the selective CCKA antagonist CAM-1481 inhibited the increase in [Ca2+]i induced by CCK-8 (half maximal inhibitory concentration = 3 nM) in GLC19 but not in H510 cells. The selective CCKB/gastrin antagonist blocked the increase in [Ca2+]i induced by CCK 8 (half-maximal inhibitory concentration = 80 pM) in H510 but not in GLC19 cells. Thus, the effects of CCK-8 are mediated through CCKA receptors in GLC19 cells and via CCKB/gastrin receptors in H510 cells. CCK-8 markedly stimulated colony formation in GLC19 cells in a dose-dependent manner in the nanomolar range, whereas over the same concentration range, gastrin had no effect on clonal growth. CAM-1481 inhibited the CCK-stimulated colony formation in GLC19 but not in H510 cells. Our results show, for the first time, that CCKA receptors can mediate Ca2+ mobilization and growth in SCLC cells and that SCLC cells express two distinct functional CCK receptor subtypes. PMID- 8221658 TI - Thymosin alpha 1 down-regulates the growth of human non-small cell lung cancer cells in vitro and in vivo. AB - The effect of thymosin alpha 1 (THN alpha 1) and its NH2-terminal fragment (THN1 14) and COOH-terminal fragment (THN15-28) on non-small cell lung cancer (NSCLC) growth was evaluated. Using an anti-THN alpha 1 antibody, receptors were identified on NSCLC cells that were pretreated with 10(-6) M THN alpha 1. [3H]Arachidonic acid was readily taken up by NSCLC cells and THN alpha 1 significantly increased the rate of arachidonic acid release. THN1-15 slightly stimulated but THN15-28 and THN beta 4 did not alter arachidonic acid release from NCI-H1299 cells. In clonogenic growth assays in vitro, THN alpha 1 (10(-6) M) significantly decreased NSCLC colony number whereas THN1-14, THN15-28, and THN beta 4 were less potent. Using growth assays in vivo, THN alpha 1 (10 micrograms s.c./day) but not THN1-14, THN15-28, or THN beta 4 inhibited significantly NSCLC xenograft formation in nude mice. These data suggest that biologically active THN alpha 1 receptors are present on NSCLC cells and that native THN alpha 1 inhibits the growth of human NSCLC. PMID- 8221659 TI - Resistance to fluorodeoxyuridine-induced DNA damage and cytotoxicity correlates with an elevation of deoxyuridine triphosphatase activity and failure to accumulate deoxyuridine triphosphate. AB - Deoxyuridine triphosphate (dUTP) misincorporation and uracil misrepair have long been implicated in fluoropyrimidine-induced DNA damage; however, the enzymatic activities responsible for these lesions have not been previously identified as critical determinants of overall sensitivity to the antitumor effects of these agents. The purpose of this study was to determine whether differences in uracil misincorporation/misrepair could account for the difference in sensitivity to fluorodeoxyuridine (FdUrd)-induced cytotoxicity and DNA damage in 2 human colorectal tumor cell lines having identical sensitivities to FdUrd-induced thymidylate synthase inhibition. Compared to HT29 cells, SW620 cells were resistant to both cytotoxicity and induction of DNA double-strand breaks, as assessed by pulse field gel electrophoresis. Alkaline elution experiments demonstrated that this resistance coincided with delayed induction of DNA single strand breaks on parental DNA and, to a lesser extent, on nascent DNA. Following treatment with FdUrd for 24 h, HT29 cells accumulated 904 +/- 273 pmol deoxyuridine triphosphate (dUTP)/10(7) cells, whereas SW620 cells accumulated 20 +/- 7 pmol dUTP. Consistent with this difference in extent of dUTP accumulation was the observation that deoxyuridine triphosphatase levels in SW620 cellular extracts were 4.4-fold higher than in HT29 extracts. The ability to accumulate dUTP, intracellular deoxyuridine triphosphatase activity, and extent of DNA damage appear to be important determinants for predicting the response to FdUrd treatment in these cell lines. PMID- 8221660 TI - Development of resistance to hydroxyurea during treatment of human myelogenous leukemia K562 cells with alpha-difluoromethylornithine as a result of coamplification of genes for ornithine decarboxylase and ribonucleotide reductase R2 subunit. AB - alpha-Difluoromethylornithine (DFMO), an enzyme-activated irreversible inhibitor of ornithine decarboxylase (ODC), was used to select two very highly drug resistant cell lines, designated K562-DFMOr and V79-DFMOr. Both DFMO-resistant cell lines exhibited elevated ODC expression due to gene amplification. Moreover, the K562-DFMOr cells, but not the V79-DFMOr cells, had an elevated level of ribonucleotide reductase subunit R2 (R2) mRNA and an increased R2 gene copy number. By analysis of their electron paramagnetic resonance spectra, an increased level of the R2 protein was observed in the K562-DFMOr cells as compared to the wild type K562 cells. This is the first description of a DFMO induced mutant cell line exhibiting coamplification of the genes for ODC and R2, and overexpression of their products. There was no coamplification of the N-myc protooncogene, which is located close to the ODC and R2 genes on human chromosome 2. The alterations exhibited by the K562-DFMOr cell line were shown to be stable for many passages and to convey resistance not only to DFMO but also to hydroxyurea, an inhibitor of ribonucleotide reductase and thus DNA replication. In the absence of the selective pressure exerted by DFMO, the V79-DFMOr cell line produced revertants by loss of ODC gene amplification within three passages. Coamplification of linked genes may turn out to be an important mechanism in the development of cross-resistance and should be considered when designing therapeutic strategies. PMID- 8221661 TI - Absence of p53 gene mutations in primary neuroblastomas. AB - Neuroblastoma is a common childhood malignancy of the sympathetic nervous system. Mutations in p53, a tumor suppressor gene located on the short arm of chromosome 17, are one of the most common genetic lesions in human cancers. The evidence for trisomies of 17q with loss of 17p in some cases of neuroblastoma led us to consider whether p53 mutations might contribute to the onset and progression of this malignancy. In this study, primary tumors from 38 neuroblastoma patients were screened for mutations within the coding exons of the p53 gene by single strand conformation polymorphism analysis, and potential mutations were further analyzed by nucleotide sequence analysis. Previously described sequence variations were detected in many of the tumors, including a silent polymorphism at codon 213 (CGA to CGG) and the nontransforming Pro to Arg substitution at codon 72 (CCC to CGC). However, no other sequence variations were detected within the coding portions of the p53 gene. This finding suggests that p53 mutations do not contribute to the etiology of neuroblastoma and that the chromosome 17 alterations observed in neuroblastoma involve genes which are distinct from the p53 locus. PMID- 8221662 TI - The "bystander effect": tumor regression when a fraction of the tumor mass is genetically modified. AB - Tumor cells expressing the herpes simplex virus thymidine kinase (HSV-TK) gene are sensitive to the drug ganciclovir (GCV). We demonstrate here that HSV-TK positive cells exposed to GCV were lethal to HSV-TK-negative cells as a result of a "bystander effect." HSV-TK-negative cells were killed in vitro when the population of cultured cells contained only 10% HSV-TK-positive cells. The mechanism of this "bystander effect" on HSV-TK-negative cells appeared to be related to the process of apoptotic cell death when HSV-TK-positive cells were exposed to GCV. Flow cytometric and electron microscopic analyses suggested that apoptotic vesicles generated from the dying gene-modified cells were phagocytized by nearby, unmodified tumor cells. Prevention of apoptotic vesicle transfer prevented the bystander effect. The toxic effect of HSV-TK-positive cells on HSV TK-negative cells was reproduced in an in vivo model. A mixed population of tumor cells consisting of HSV-TK-positive and HSV-TK-negative cells was inoculated s.c. into mice. Regression of the tumor mass occurred when the inoculum consisted of as few as 10% HSV-TK-expressing tumor cells. The bystander effect was also demonstrated in i.p. tumor studies. Initial experiments demonstrated that prolonged survival (> 70 days) occurred when a mixture containing 50% HSV-TK positive and 50% HSV-TK-negative cells was injected i.p. followed by GCV treatment. Further, survival was prolonged for mice with a preexisting HSV-TK negative i.p. tumor burden by injecting HSV-TK-positive cells and GCV. These results suggest that genetic modification of tumor cells may be useful for developing cancer therapies. PMID- 8221663 TI - Mutations of the p53 gene are involved in Ewing's sarcomas but not in neuroblastomas. AB - We have investigated the frequency of p53 gene mutations in Ewing's sarcoma (ES) and neuroblastoma (NB) by using polymerase chain reaction-single strand conformation polymorphism analysis for genomic DNA or complementary DNA generated from total RNA. Mutations of the p53 gene were found in six of seven ES cell lines: a missense mutation of TGC (Cys)-->TAC (Try) at codon 141 in one, a missense mutation of CGT (Arg)-->TGT (Cys) at codon 273 in one, a missense mutation of TGC (Cys)-->TTC (Phe) at codon 176 in three, and one base deletion of CGC-->CG at codon 283 in one. Further analysis of 14 ES and related primary tumors showed mutations of the p53 gene in only two: one base insertion of CCG- >CCCG at codon 152 in one and a missense mutation of GGC (Gly)-->GTC (Val) at codon 154 in the other. Both of the two tumors were obtained from patients with an advanced stage disease. Three of the eight ESs with mutations of the p53 gene showed the same missense mutation at codon 176, suggesting the mutational hot spot of the p53 gene in ESs. In contrast to ES, none of 6 NB cell lines or 48 NB tumors including advanced-stage ones with or without N-myc amplification showed any aberration of the p53 gene. Our findings suggest that mutations of the p53 gene in ES might represent late genetic events related to tumor progression, and that aberrations of the p53 gene might not be involved in the development or the progression of NB. PMID- 8221664 TI - Basic fibroblast growth factor secreted by an animal tumor is detectable in urine. AB - Basic fibroblast growth factor (bFGF or FGF-2) is abnormally elevated in the serum and urine of patients with many types of cancer. However, the source of the bFGF is unclear. We developed a model that could distinguish between tumor derived and host-derived bFGF. We gave athymic mice s.c. injections of cells of the murine K1000 tumor, which secretes a bFGF mutein (bFGF CS23) as its dominant angiogenic factor. Controls were given injections of Lewis lung carcinoma or saline. Urine was collected for 9 weeks, and bFGF was quantitated using two immunoassays which can discriminate between tumor bFGF CS23 and native bFGF. None of the mice had detectable urinary native bFGF, and no control mice had detectable urinary bFGF CS23. In contrast, urine from mice bearing the K1000 tumor revealed detectable bFGF CS23 by 2 weeks, and bFGF CS23 increased with increasing tumor volume throughout the study. Because bFGF CS23 is not produced by other cells, the bFGF CS23 in the urine most likely came from the K1000 tumor and not from the host. These results suggest that the source of elevated bFGF in the urine of human cancer patients is, at least in part, the tumor itself. PMID- 8221665 TI - Epidermal growth factor receptor-mediated autocrine and paracrine stimulation of human transitional cell carcinoma. AB - Stromal-epithelial interactions may play a key role in tumor growth and metastasis. We have established a model to study the cellular and molecular basis of this paracrine interaction both in vivo and in vitro using a human transitional cell carcinoma cell line (WH). s.c. coinoculation of 1 x 10(6) WH cells with 1 x 10(6) nontumorigenic fetal rat urogenital sinus mesenchymal (rUGM) cells in athymic mice accelerated carcinoma growth 20 times faster than isolated WH cell inoculations and 4 times faster than coinoculations of the same number of NIH-3T3 or human bladder fibroblasts. Characterization of these chimeric tumors with immunohistochemical and DNA dot-blot analyses documented their predominantly human component. To evaluate the underlying mechanisms involved in this paracrine mediated in vivo tumor growth acceleration, Northern analyses for growth factors (GFs) and extracellular matrix (ECM) expression in the different cell lines, as well as in vitro mitogenic assays, were performed. Northern analysis revealed basic fibroblast growth factor, transforming growth factor alpha, and epidermal growth factor receptor expression by WH cells but not rUGM cells; ECM components (fibronectin and collagens I and IV) were expressed only in the fibroblast cell lines. Cell type-specific paracrine growth factors are produced by cultured stromal and epithelial cells with a 2-3-fold bidirectional increase in WH and rUGM cell growth when cultured with reciprocal cell-type conditioned medium. An autocrine growth loop was observed for WH but not rUGM cells. WH cell growth is stimulated in vitro by low concentrations of transforming growth factor alpha and epidermal growth factor, while rUGM cell growth is stimulated 3-fold by basic fibroblast growth factor. Antiepidermal growth factor receptor antibodies completely inhibited autocrine and paracrine pathways stimulating WH cell growth, while anti-basic fibroblast growth factor antibodies had no inhibitory effect. These observations suggest that autocrine and paracrine growth factor stimulation of WH bladder carcinoma cell growth is most likely mediated by an epidermal growth factor receptor-related pathway. The predominant expression of ECM by fibroblasts in this model suggests that stromal cell ECM components may modulate tumor cell growth and angiogenesis possibly through mechanisms involving cellular adhesion, chemotaxis, or growth factor action. PMID- 8221666 TI - Tumor cell heme uptake induces ferritin synthesis resulting in altered oxidant sensitivity: possible role in chemotherapy efficacy. AB - Neovascularization and hemorrhage are common features of malignant tumors. We wondered whether hemoglobin derived from extravasated RBC deposits heme-derived iron into the tumor, which could modulate the sensitivity of cancer cells to oxidant-mediated injury. A brief exposure (1 h) of 51Cr-radiolabeled breast cancer cells (BT-20) but not colon cancer cells (Caco-2) to hemin (10 microM) or FeSO4 (10 microM) significantly enhances cytotoxicity mediated by 0.5 mM hydrogen peroxide (H2O2). Associated with Caco-2 resistance, these cells were found to be enriched in the endogenous iron chelator, ferritin. If cellular ferritin is even further increased through 1 h incubation (24 h prior to H2O2 exposure) of both cell types with hemin, FeSO4, or exogenous spleen apoferritin itself (24 h), marked resistance to H2O2-mediated cytotoxicity is manifest. Under several conditions, the sensitivity of tumor cells to oxidant-mediated lysis is inversely proportional to their ferritin content. Pretreatment of BT-20 and Caco-2 cells with hemin or FeSO4 rapidly increases H-ferritin mRNA but only slightly increases L-ferritin mRNA; nevertheless, large increases in overall ferritin content of iron-exposed cells result. Data analogous to those with H2O2-mediated cytotoxicity were obtained in studies of bleomycin-engendered DNA strand breakage and cell damage, i.e., brief treatment of BT-20 cells with both hemin or FeSO4 significantly increases their sensitivity to bleomycin (100 micrograms/ml), whereas treatment followed by 24 h incubation with media alone significantly protects against bleomycin toxicity. We speculate that acute exposure of tumors to iron (e.g., derived from heme-proteins in hemorrhagic cancerous lesions) may increase sensitivity of some cancer cells, particularly those relatively low in endogenous ferritin, to oxidant-mediated lysis. In contrast, repeated, more chronic, exposure effector cells or chemotherapeutic agents, an effect derived from their increased synthesis and accumulation of the intracellular iron scavenger, ferritin. PMID- 8221667 TI - Apoptosis induced in Burkitt's lymphoma cells via Gb3/CD77, a glycolipid antigen. AB - Gb3/CD77 is a glycolipid antigen, specifically expressed on two different B-cell populations, Burkitt's lymphoma and a subset of tonsillar B-lymphocytes located in germinal centers, which could be the normal counterpart of Burkitt cells. Both Gb3/CD77(+) populations have recently been shown to enter programmed cell death (apoptosis) readily. Here we show that verotoxin, also called Shiga-like toxin, which is known to bind to the carbohydrate moiety of Gb3/CD77, induces cell death in Gb3/CD77(+) Burkitt's lymphoma cells, not only by inhibiting protein synthesis as classically described but also through an additional mechanism, namely apoptosis. Furthermore a recombinant B-subunit of verotoxin, which carries only the binding property of the holotoxin, also induces apoptosis in Gb3/CD77(+) cells. Gb3/CD77 could thus represent the first example of a glycolipid antigen able to transduce a signal leading to apoptosis. PMID- 8221668 TI - Coexistence of aneuploid subclones within a myeloma cell line that exhibits clonal immunoglobulin gene rearrangement: clinical implications. AB - A new human myeloma cell line, ANBL-6, was established and characterized at the genotypic and phenotypic levels. The cells exhibit a clonally rearranged immunoglobulin gene locus and resemble plasma cells morphologically. The ANBL-6 cells also exhibited an absolute dependence on exogenous interleukin 6 for growth. Of interest, DNA ploidy analysis suggested the existence of a near diploid as well as a near-tetraploid population in this cell line. Cytogenetic studies confirmed the existence of two aneuploid karyotypes and further revealed a clonal relationship between the two karyotypes, as evidenced by numerous shared structural abnormalities. To determine whether the near-diploid cells functioned as stem cells for the near-tetraploid population, the near-diploid population was separated via flow cytometry and recultured prior to ploidy analysis. This population was observed to remain predominantly near-diploid over time, suggesting that these cells did not function as stem cells for the near tetraploid population. However, the near-tetraploid cells did exhibit a growth advantage in vitro. Moreover, sequential ploidy analysis performed retrospectively on fresh bone marrow cells from the patient also suggested that there was an expansion of the near-tetraploid population during clinical relapse. These results suggest that both populations are self-regenerating and reflect the consequences of clonal evolution in the myeloma tumor. The coexistence of clonally related subclones with shared chromosomal abnormalities, however, suggests that the near-tetraploid subclone was derived from the near-diploid subclone at an unknown time during tumorigenesis. PMID- 8221669 TI - Malignant transformation of cutaneous lesions in renal allograft patients: a role for human papillomavirus. AB - Renal allograft recipients are at greatly increased risk of developing squamous cell carcinomas. As these frequently arise adjacent to areas of multiple viral warts, we have investigated a possible role for human papillomavirus in malignant transformation within this population. We established, as primary cultures, keratinocytes from 24 lesions of varying degrees of squamous atypia from 9 patients. Ten of 14 cultures screened for the presence of episomal human papillomavirus DNA were positive using a mixed probe for cutaneous human papillomaviruses, although episomal copy was universally lost with continued passage. Three cultures, 2 of which were derived from malignant tissue and 1 from a benign lesion, were positive when screened with a probe for potentially oncogenic human papillomavirus DNAs 5 or 8. Both positive cultures of malignant origin exhibited extended lifespan and have been briefly characterized by morphology and growth requirements. PMID- 8221670 TI - Systematic HRAS amplification in ovary-independent mammary tumors: correlation with progressively anaplastic phenotypes. AB - Progression of well differentiated rat mammary adenocarcinomas to very anaplastic phenotypes was found to correlate with a systematic and significant amplification of a mutant HRAS allele. Tumors with high amplification levels of this oncogene were analyzed by chromosomal in situ hybridization; in four of the cases the amplified sequences did not reside at the native chromosome 1 locus but were localized in a novel marker chromosome. The model described has potential as a reproducible system for the study of the chromosomal and cellular mechanisms operative "in vivo" for oncogene amplification. PMID- 8221671 TI - Sequence-specific interaction of a conformational domain of p53 with DNA. AB - Mutations within a conserved "conformational" domain of the p53 protein have frequently been observed in a wide variety of human cancers. A hybrid protein containing the wild-type conformational domain of p53 fused to protein A bound to calf thymus DNA and a specific p53 DNA-binding motif. Hybrid proteins containing mutations in p53 bound to DNA less efficiently than wild-type hybrid protein. In addition, competition experiments showed that mutated p53 DNA-binding motif failed to interact with p53 hybrid proteins. The DNA-binding activity of wild type p53 hybrid protein was inhibited by the metal chelator 1,10-phenanthroline. These results demonstrate that DNA-binding activity resides in the conformational domain of p53, providing a structural model for disruption of DNA binding by mutation. Furthermore, metal ions may regulate binding of p53 to DNA by modulating its conformation. PMID- 8221672 TI - Levels of matrix metalloproteases in bladder cancer correlate with tumor grade and invasion. AB - We have used quantitative zymography to measure levels of the type IV collagenases matrix metalloproteinase (MMP)-9 and MMP-2 in 42 biopsies of transitional cell carcinoma and in 7 biopsies of normal bladder. Mean levels of MMP-9 were significantly higher in tumor compared with normal samples (P = 0.08). Levels of MMP-9 and active MMP-2 increased with tumor grade (test for trend, P = 0.002 and P = 0.05, respectively). Levels of MMP-9 and activated MMP-2 were also higher in invasive tumors than in superficial ones (P = 0.001 and P = 0.008, respectively). In situ hybridization studies showed that the mRNAs for both MMP-2 and MMP-9 were located chiefly in the stroma rather than epithelial tumor cells and were concentrated at the interface between the two tissues. PMID- 8221673 TI - Poly(ADP-ribose) polymerase can bind melphalan damaged DNA. AB - As a means of identifying damage recognition proteins involved in repair of nitrogen mustard lesions in chronic lymphocytic leukemia, we performed Southwestern analysis using a probe damaged with melphalan and protein extracts from chronic lymphocytic leukemia patients. We detected proteins with molecular weights of 116,000, 66,000, and 64,000 which bound the damaged probe with a higher specificity than the undamaged probe. The M(r) 66,000 and 64,000 proteins were determined to be degradation products of the M(r) 116,000 protein. The M(r) 116,000 protein was identified as poly(ADP-ribose) polymerase. The use of methoxyamine, an inhibitor of DNA strand breakage following depurination, significantly reduced binding of the melphalan damaged probe to poly(ADP-ribose) polymerase. Following depletion of poly(ADP-ribose) polymerase from the cell extracts, no other binding activity was discovered. Thus, poly(ADP-ribose) polymerase is the only demonstrable protein in chronic lymphocytic leukemia cells which can bind to a DNA probe damaged with melphalan. PMID- 8221674 TI - Synthetic retinoid fenretinide is effective against a human ovarian carcinoma xenograft and potentiates cisplatin activity. AB - Fenretinide or N-(4-hydroxyphenyl) retinamide (4HPR) is a synthetic retinoid currently being tested clinically, which can inhibit the development and the growth of breast and prostate cancers in rodents. The efficacy of 4HPR alone and in combination with cisplatin was tested against the human ovarian carcinoma IGROV-1 xenograft i.p. Administration p.o. of 4HPR was not effective, whereas intracavitary treatment significantly increased the survival time of treated mice. It also enhanced the antitumor activity of cisplatin. These findings suggest that 4HPR may be an active agent against epithelial ovarian tumors. PMID- 8221675 TI - DNA strand bias in the repair of the p53 gene in normal human and xeroderma pigmentosum group C fibroblasts. AB - We have measured the gene-specific and strand-specific DNA repair of UV-induced cyclobutane pyrimidine dimers in the p53 tumor suppressor gene in a normal, repair-proficient human fibroblast strain and in fibroblasts from a patient with the repair deficient disorder xeroderma pigmentosum, complementation xeroderma pigmentosum group C (XP-C). In both cell strains, repair was measured in the p53 gene and in its individual DNA strands. For comparison, the repair also was measured in other genomic regions in these human fibroblast strains, including the housekeeping gene dihydrofolate reductase, and two inactive genomic regions, the delta globin gene, and the 754 locus of the X chromosome. In both cell strains, we find that the p53 gene is repaired faster than the dihydrofolate reductase gene and much more efficiently than the inactive genomic regions. Selective repair of the transcribed DNA strand of p53 is observed in both human cell strains; the strand bias of repair is particularly distinct in XP-C. Mutations specific to the nontranscribed strand may occur due to replication errors at the sites of unrepaired DNA damage. Therefore, our results predict that the majority of mutations in skin cancers, especially those from patients with XP C, would occur on the nontranscribed strand of the p53 gene. Indeed, Dumasz et al. (Proc. Natl. Acad. Sci. USA, in press, 1993) report such a strand bias of p53 mutation in skin cancers from XP-C patients. PMID- 8221676 TI - Biophysical characterization of one-, two-, and three-tandem repeats of human mucin (muc-1) protein core. AB - Until recently mucin tandem repeat protein cores were believed to exist in random coil conformations and to attain structure solely by the addition of carbohydrates to serine and threonine residues. Matsushima et al. (Proteins Struct. Funct. Genet., 7: 125-155, 1990) recently proposed a model of the secondary structure of proline rich tandem repeat proteins that has challenged this idea, especially for the case of the human polymorphic epithelial mucin encoded by the muc-1 gene. We report here results of structural analyses of the muc-1 protein core by using synthetic peptide analogues. Synthetic peptides were prepared to correspond to one-, two-, and three-tandem repeats of muc-1. Results of one- and two-dimensional 1H NMR correlation spectroscopy on these peptides confirm that the muc-1 protein core is not a random-coil secondary structure. Long-lived amide protons are protected in D2O, and increasing spectral complexity in the region of the beta-protons of Asp2 and His 15 reveals that structural changes are occurring as the number of repeats increases. The greatest changes occur when the number of repeats increases from one to two. These results are supported by the reactivity of a panel of monoclonal antibodies raised against tumor associated muc-1 with these synthetic peptides in enzyme-linked immunosorbent assay. The primary immunodominant mucin epitope, PDTRP, does not appear to attain a native conformation in the single repeat peptide (20 amino acids, starting with P), but is expressed on peptides with multiple repeats. Intrinsic viscosity measurements of the peptide containing three repeats indicate that an ordered structure present in solution is rod shaped. The circular dichroism spectrum of the same peptide is dominated by proline in the trans conformation. These results are all consistent with the prediction that the muc-1 tandem repeat polypeptide core forms a polyproline beta-turn helix. PMID- 8221677 TI - Genetic and enzymatic basis for the differential expression of GM2 and GD2 gangliosides in human cancer cell lines. AB - Using beta 1,4-N-acetylgalactosaminyltransferase (EC 2.4.1.92) complementary DNA, the correlation of gene expression, enzyme activity, and expression of ganglioside antigens was analyzed in 20 human tumor cell lines. In many lines, GM2 and/or GD2 were the most complex structures examined. Northern blot analysis revealed 5.2- and 3.0-kilobase mRNAs in almost all cell lines expressing GD2 and/or GM2. Some melanoma lines, however, showed no bands although they expressed fairly high levels of GD2. These cell lines expressed very high levels of alpha 2,8-sialyltransferase and the resulting product, GD3. Semiquantitative RT-PCR demonstrated that even cell lines with no bands in Northern blot contained 0.4 2.5% of mRNA level in the highest expressing cell line. These results indicate that GD2 expression on individual cell lines is regulated not only by the expression level of the N-acetylgalactosaminyl transferase but also by the amount of its precursor structure (GD3) and alpha 2,8-sialyltransferase present in the cells. beta 1,4-N-acetylgalactosaminyltransferase activities and mRNA levels generally correlated quite closely. A few lines, however, showed lower enzyme activities than expected from their mRNA levels, indicating the possibility that the enzyme is being regulated by translational or posttranslational modification such as phosphorylation and glycosylation as well as by transcriptional regulation. Depending on their patterns of ganglioside synthesis and expression, the lines examined were classified into 6 groups which were characteristic of different tumor cell types. PMID- 8221678 TI - Folate deficiency increases genetic damage caused by alkylating agents and gamma irradiation in Chinese hamster ovary cells. AB - The effect of folate deficiency on genetic damage caused by alkylating agents and gamma-irradiation was studied in Chinese hamster ovary (CHO) cells. Mutant frequencies of 6-thioguanine-resistant and diphtheria toxin-resistant cells were not significantly increased by incubation in low-folate medium. In contrast, folate deficiency increased the mutant frequencies of 6-thioguanine-resistant cells caused by N-ethyl-N-nitrosourea or ethyl methanesulfonate by about 3-fold. Folate deficiency was associated with a 70% increase of diphtheria toxin resistant cells after exposure to ethyl methanesulfonate. Folate deficiency alone caused DNA strand breaks equivalent to 26 cGy, as monitored by alkaline filter elution. Following 400 cGy of gamma-irradiation, folate-deficient cells manifested strand breaks equivalent to a dose of 710 cGy. CHO cells in folate containing medium repaired breaks within 3 h, while cells in low-folate medium had an increased break frequency (P = 0.02) at 3 h and were unable to fully repair radiation-induced damage even after 9 h. These studies indicate that folate deficiency acts synergistically with alkylating agents to increase somatic mutation and with gamma-irradiation to promote DNA strand breaks in CHO cells. Folate deficiency appears to potentiate the genetic damage caused by mutagens/carcinogens by limiting DNA repair. PMID- 8221679 TI - Protection against malignant conversion of chemically induced benign skin papillomas to squamous cell carcinomas in SENCAR mice by a polyphenolic fraction isolated from green tea. AB - Progression of benign tumors to malignant cancer is critical since cancerous lesions are capable of metastatic spread and eventually causing death. Inhibitors of the conversion process, therefore, would likely be useful as cancer chemopreventive agents. In this study, we assessed the protective effect of topical application of a polyphenolic fraction isolated from green tea (GTP) against spontaneous as well as benzoyl peroxide (BPO)- and 4-nitroquinoline-N oxide (4-NQO)-enhanced malignant conversion of chemically induced skin papillomas in SENCAR mice. Papillomas were induced in SENCAR mice by topical application of 7,12-dimethylbenz(a)anthracene as a tumor-initiating agent followed by twice a week application of 12-O-tetradecanoylphorbol-13-acetate as a tumor-promoting agent. Beginning at the 20th week, when papilloma yield was stabilized, enhanced malignant conversion was achieved by twice weekly topical application of either BPO or 4-NQO, whereas spontaneous malignant conversion was associated with topical application of acetone. In these protocols, preapplication of GTP (6 mg/animal) 30 min prior to skin application of acetone, BPO, or 4-NQO resulted in 14, 31, and 29% protection, respectively, in terms of percentage of mice with carcinomas, and 20, 35, and 43% protection in terms of number of carcinomas/mouse. In these experiments, a BPO- and 4-NQO-enhanced rate of malignant conversion was also found to be decreased significantly by the skin application of GTP; however, such effects of GTP were less profound in the cases of spontaneous malignant conversion. The results of this study suggest that, in addition to its chemopreventive effects against tumor initiation and promotion stages of multistage carcinogenesis, green tea also possesses significant protective effects against tumor progression, specifically tumor progression induced by BPO and 4-NQO. PMID- 8221680 TI - Kinetics and tissue distribution of the radiolabeled chimeric monoclonal antibody MOv18 IgG and F(ab')2 fragments in ovarian carcinoma patients. AB - Twenty-four patients suspected of having ovarian carcinoma received i.v. injection with a combination of radiolabeled intact IgG (1 mg) and F(ab')2 fragments (1 mg) of the chimeric monoclonal antibody MOv18, each form labeled with 1.85 MBq 131I or 125I. Laparotomy was performed either 2 or 6 days after injection, and the uptake of radioactivity was determined in a total of 329 biopsies of normal and malignant tissues. The mean elimination half life in plasma of cMOv18 IgG and F(ab')2 was 70 +/- 8 (SD) and 20 +/- 5 h, respectively. The mean uptake of IgG in tumor biopsies was 3.6-fold higher two days after injection and 6.9-fold higher than the uptake of F(ab')2 6 days after injection. Uptake in normal tissues was 3.3 and 5.5 times higher for IgG at 2 and 6 days, respectively. Two days after injection, the mean ratio of the uptake in tumor:normal tissue/patient was 3.8 +/- 1.5 and 4.0 +/- 1.8 for radiolabeled cMOv18 IgG and F(ab')2, respectively. Six days after injection, this was 6.7 +/- 4.7 for Ig G and 5.7 +/- 4.1 for F(ab')2. cMOv18 IgG has a longer circulation time in blood, a higher uptake in tumor and normal tissues, and a longer retention time compared to the F(ab')2 fragments. However, the tumor:normal tissue ratios are similar. The results do not warrant a definite conclusion as to which antibody form is most suitable for therapeutic application of antibodies but provide a more firm basis for rational design of therapeutic targeting studies using immunoconjugates. PMID- 8221681 TI - Cytokinetic effects of interferon in colorectal cancer tumors: implications in the design of the interferon/5-fluorouracil combinations. AB - Interferon (IFN) has been shown to enhance the cytotoxic effects of 5 fluorouracil (5FUra) in colorectal cancer, and clinical trials with this combination resulted in higher response rate with respect to 5FUra alone. IFN is generally administered s.c. three times a week. This prolonged exposure could determine a block of tumor cells in the G0-G1 phase of the cell cycle, thus rendering tumor cells insensitive to 5FUra, an S-phase specific agent. In order to verify the presence of this block, 21 operable colorectal cancer patients were treated with IFN-alpha 2b at the dose of 3 megaunits every other day in the week before operation, while another 22 represented the control group. Samples of tumor tissue were taken at endoscopy and operation. [3H]Thymidine labeling index and flow cytometry were used to assess the S-phase fraction. In IFN treated patients, we found a significant statistical difference between the mean percentage of S-phase fractions evaluated either by labeling index (P = 0.00001) or by flow cytometry (P < 0.001). On the contrary, this difference was not present in the control group: labeling index, P = 0.06; flow cytometry, P = 0.08. Furthermore a significant increase in the G0-G1 phase of the cell cycle was found after IFN administration (P < 0.001) but not in the control group. Our results suggest that IFN reduces the S-phase fraction in colorectal cancer tumors. This action should be considered in the design of the 5FUra/IFN combination because it could decrease 5FUra activity, leading to a loss or a decrease in the advantage of 5FUra modulation by IFN. PMID- 8221682 TI - Dihydropyrimidine dehydrogenase activity in human peripheral blood mononuclear cells and liver: population characteristics, newly identified deficient patients, and clinical implication in 5-fluorouracil chemotherapy. AB - Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme in the catabolism of 5-fluorouracil (FUra), one of the most widely used anticancer drugs. Previous studies from our laboratory demonstrated the clinical importance of DPD in cancer patients (G. D. Heggie, J-P. Sommadossi, D. S. Cross, W. J. Huster, and R. B. Diasio. Cancer Res., 47: 2203-2206, 1987; B. E. Harris, R. Song, S-j. Soong, and R. B. Diasio. Cancer Res., 50: 197-201, 1990), particularly in those with DPD deficiency who experience severe FUra toxicity (including death) following FUra treatment [R. B. Diasio, T. L. Beavers, and J. T. Carpenter. J. Clin. Invest., 81: 47-51, 1988; B. E. Harris, J. T. Carpenter, and R. B. Diasio. Cancer (Phila.), 68: 499-501, 1991]. We now suggest that measurement of DPD activity may be useful in routine screening of cancer patients prior to FUra treatment. In this paper, we describe the following serial studies: (a) we developed a sensitive, accurate, and precise DPD assay and a storage method to stabilize DPD activity, permitting large scale DPD screening in cancer patients; (b) we demonstrated a normal distribution (Gaussian distribution) of human DPD activity from peripheral blood mononuclear cells (PBM-DPD) in a population study. Baselines for PBM-DPD with fresh and frozen samples were 0.425 +/- 0.124 (SD) and 0.189 +/- 0.064 nmol/min/mg protein, respectively. The 95% and 99% distribution ranges for both fresh and frozen samples were also determined, providing criteria for detection of DPD-deficient patients; (c) we identified nine new patients with profound or partial DPD deficiency; (d) we determined a baseline for human liver DPD activity, which was shown to be 0.360 +/- 0.182 nmol/min/mg protein (frozen samples); (e) we did a preliminary evaluation of liver DPD from deficient patients. Low liver DPD activity in two deficient patients correlated with low PBM-DPD activity. Using a polyclonal antibody raised against human liver DPD in our laboratory (Z. Lu, R. Zhang, and R. B. Diasio. J. Biol. Chem., 267: 17102-17109, 1992), Western blot analysis demonstrated decreased DPD protein in the liver cytosol from DPD-deficient patients compared to normal subjects. These results may be useful in improving the effectiveness and/or lessening the toxicity of FUra chemotherapy. PMID- 8221683 TI - High affinity binding and direct antiproliferative effects of LHRH analogues in human ovarian cancer cell lines. AB - Recently, specific binding sites for luteinizing hormone releasing hormone (LHRH) and its analogues have been demonstrated in biopsy samples of human epithelial ovarian cancer. Their biological significance remained obscure. In this study we ascertained whether such LHRH-binding sites are also present in the human epithelial ovarian cancer cell lines EFO-21 and EFO-27 and if they could mediate antiproliferative effects of LHRH analogues. Using [125I, D-Trp6]LHRH, a high affinity/low capacity binding site was detected in both lines: EFO-21 (Kd1 = 1.5 x 10(-9) M; binding capacity (Bmax1) = 4.9 fmol/10(6) cells) and EFO-27 (Kd1 = 1.7 x 10(-9) M; Bmax1 = 3 fmol/10(6) cells). In addition, a second class of low affinity/high capacity binding sites (EFO-21: Kd2 = 7.5 x 10(-6) M; Bmax2 = 24 pmol/10(6) cells; EFO-27: Kd2 = 4.3 x 10(-6) M; Bmax2 = 14.5 pmol/10(6) cells) was demonstrated. Specific binding of [125I, D-Trp6]LHRH was displaced with nearly equal efficiency by unlabeled [D-Trp6]LHRH, the LHRH-antagonists SB-75 and Hoe-013, and by native LHRH but not by unrelated peptides such as oxytocin and somatostatin. In the presence of 10(-5) M agonist [D-Trp6]LHRH, the proliferation of both cell lines was significantly reduced to 77% of controls after 24 h and to approx. 60% after 6 days. Lower concentrations (10(-9) M) of the agonist, significantly decreased the proliferation to 87.5% for EFO-21 and 86% for EFO-27 after 6 days. These antiproliferative effects were enhanced by increasing doses of [D-Trp6]LHRH and were maximal at 10(-5) M (EFO-21: 65.5% of control, EFO-27: 68% of control). Similar dose-dependent antiproliferative effects were obtained in EFO-21 line with the LHRH-antagonists SB-75 and Hoe-013, while these analogues had no effects on the proliferation of EFO-27 cells. SB-75 partly antagonized the antiproliferative effect of [D-Trp6]LHRH in a dose dependent way in the EFO-27 line. These data suggest that LHRH analogues can directly inhibit the in vitro proliferation of human ovarian cancer cells. This effect might be mediated through the high affinity LHRH binding sites. PMID- 8221684 TI - Localization of estrogen receptors in interstitial cells of hamster kidney and in estradiol-induced renal tumors as evidence of the mesenchymal origin of this neoplasm. AB - The mechanism of estrogen-induced and -dependent kidney carcinogenesis in Syrian hamsters and the cell of origin of the tumor are not well understood; they have been investigated in this study by mapping the cellular locations of estrogen receptor (ER) in estrogen-dependent tumors, in kidney tissue of hamsters treated with estradiol for 0.5 and 5.5 months, and in kidneys of age-matched controls. To validate the methods used, receptors have also been localized in uteri of hamsters and rats and in female hamster kidneys. ERs have been identified in cryostat sections by immunocytochemical techniques using an affinity-purified ER antibody, ER-715. Nuclei of tumors were intensely stained for ERs. In estrogen treated kidneys and in controls, ER protein was identified in interstitial cells and capillaries, in arteries, and in renal corpuscles, particularly in podocytes and in the parietal layers surrounding the renal corpuscles. There was no ER protein in tubular epithelia even when tubuli were surrounded by tumor cells. The ER distribution in female hamster kidneys closely matched that in male kidneys. However, the staining intensity was stronger in female than in male kidneys. In hamster uteri, there was an intense ER-positive reaction in the nuclei of stroma, in stromal vessels, and in the luminal epithelia as demonstrated previously by others in rat uteri. ER mRNA has also been demonstrated by Northern blot analysis in estrogen-treated kidneys which contained tumors but was undetectable in untreated kidneys. The localization of ERs in estrogen-dependent tumors and in interstitial cell types but not in tubular epithelia supports previous conclusions of an interstitial origin of estrogen-induced hamster kidney tumors. PMID- 8221685 TI - Increased growth rate and tumor burden of spontaneously metastatic Walker 256 cancer cells in the skeleton of bisphosphonate-treated rats. AB - We have studied the effect of 3-amino-1-hydroxypropylidene-1,1-bisphosphonate (APD) on the morphology of rat bone and the metastatic behavior of Walker 256 (W256) cancer cells in the rat skeleton. Male Fischer rats (150-175 g) received s.c. injections for 7 days with APD (0.5 mg/kg body weight/day) (+ APD; n = 20) or with vehicle (-APD; n = 20). Subsequently, 10 + PD and 10 -APD rats received i.m. injections with W256 cells (+ W256), and the remaining rats received injections of vehicle (-W256). All rats were killed 14 days later. Trabecular bone volume was increased by 46 +/- 3% by APD treatment alone and was decreased by 56 +/- 7% (SEM) by W256 tumor burden alone. After 14 days of tumor burden, + APD/+ W256 rats had 3-fold more trabecular bone than did -APD/+W256 rats. Despite this bone-sparing effect, APD treatment of +W256 rats was associated with a 2.6 fold increase in skeletal tumor burden, while metastatic tumor burden in the liver, lungs, and kidneys was unaffected. The increased skeletal tumor burden in + APD/+ W256 rats was accompanied by an increase in the growth rate of W256 cells located in bone. Independent of APD treatment, W256 cells located adjacent to trabecular bone surfaces had greater growth rates than did W256 cells in the marrow, located > 50 microns from trabecular bone. In summary, the APD-induced increase in trabecular bone volume in rats is associated with a selective increase in skeletal tumor burden and an increased growth rate of W256 cells in the skeleton. PMID- 8221686 TI - Cigarette smoking is a determinant of DT-diaphorase gene expression in human non small cell lung carcinoma. AB - The levels of NAD(P)H:(quinone-acceptor) oxidoreductase (EC.1.6.99.2) (DT diaphorase) mRNA and enzyme activity have been studied in paired human normal lung and non-small cell lung tumor samples from patients with a history of cigarette smoking. There were significantly higher levels of DT-diaphorase mRNA (1.2 kilobases) in lung tumor compared to normal lung tissue of patients who had stopped smoking more than 6 months before surgery, with relative values (normalized to beta-actin mRNA) of 29.6 +/- 7.8 (SE) in the lung tumor compared to 11.7 +/- 2.2 in normal lung tissue (P < 0.05). There was no significant difference in DT-diaphorase mRNA between lung tumor and normal lung tissue of subjects who were smokers at the time of surgery, with values of 16.5 +/- 2.1 and 15.3 +/- 2.5 (P > 0.05), respectively. DT-diaphorase enzyme activity in normal and tumor lung tissue was positively correlated with DT-diaphorase mRNA (r = 0.908, P < 0.01). The results of the study suggest that DT-diaphorase does not function as an inducible protectant enzyme in human lung against oxidant species and carcinogens present in cigarette smoke. Metabolism of some anticancer drugs by DT-diaphorase can alter their activity. Differences in DT-diaphorase between lung tumors of smokers and past smokers might alter the response to these drugs. PMID- 8221687 TI - Kinetics and mechanism of mitomycin C bioactivation by xanthine dehydrogenase under aerobic and hypoxic conditions. AB - These studies examined the kinetic and mechanistic parameters of mitomycin C (MMC) bioreduction by xanthine dehydrogenase (XDH), an enzyme recently shown to be capable of MMC activation. The bioreduction of MMC by XDH leads to the formation of 2,7-diaminomitosene (2,7-DM) under both aerobic and hypoxic conditions, with greater 2,7-DM formation observed under hypoxic conditions. The XDH-induced formation of 2,7-DM is pH dependent with increasing formation as the pH is varied from 7.4 to 6.0. In this study, the kinetics of MMC bioreduction by XDH was assessed under aerobic and hypoxic conditions and at pH 7.4 and 6.0. MMC interaction with XDH was also assessed by monitoring the ability of MMC to inhibit XDH-mediated uric acid and NADH formation. The ability of xanthine to serve as reducing equivalents for MMC reduction was also measured. Aerobically but not hypoxically, MMC reduction by XDH followed Michaelis-Menten kinetics. Kinetic constants calculated under aerobic conditions suggested that the pH dependent increase (pH 6.0 > pH 7.4) in MMC activation by XDH is due to an approximately 2-fold decrease in the Km and a 2-fold increase in the Vmax at pH 6.0. Stimulation of uric acid formation and decreases in NADH formation by XDH in the presence of MMC suggest that MMC interaction with XDH may occur at the NAD(+) binding region of the enzyme. The ability of xanthine to serve as reducing equivalents for MMC conversion to 2,7-DM also supports the hypothesis that MMC reduction is occurring at the NAD+ site. PMID- 8221688 TI - Growth hormone administration preserves lean body mass in sarcoma-bearing rats treated with doxorubicin. AB - Cachexia and malnutrition play a significant role in the morbidity associated with antineoplastic chemotherapy regimens. Conventional nutritional support during cancer therapy has shown little benefit in terms of reducing morbidity and mortality. We examined the anabolic properties of growth hormone (GH) that preserve normal body composition in sarcoma-bearing animals treated with doxorubicin. On day 0, 40 male Fischer 344 rats were inoculated with 10(6) methylcholanthrene-induced sarcoma cells s.c. in the left flank. On day 9, animals were randomized into 3 groups: control (CTL, n = 13); doxorubicin (DOX, n = 13); and doxorubicin plus GH (DOX-GH, n = 14). Two CTL animals were excluded due to tumor invasion into the peritoneal cavity. From day 9 to day 23, the DOX GH group received daily s.c. recombinant rat GH injection (1 mg/kg). On day 13, DOX and DOX-GH groups received 7 mg/kg of DOX i.v. while the CTL group received sham i.v. sterile saline injection. Body weight and tumor dimensions were measured daily. On day 23, all animals were weighed and sacrificed. Tumors were resected and weighed. Body composition analysis was performed. Plasma GH levels were measured by radioimmunoassay and insulin growth factor 1 levels were measured by chondrocyte proliferation bioassay. The DOX-GH group had a significantly higher mean body weight, carcass weight, total fat free body mass, insulin growth factor 1, and GH plasma levels as compared to the DOX group. No difference in total food intake was observed between the DOX and DOX-GH groups. There was no difference in final tumor weight and tumor growth rate between DOX and DOX-GH groups. Exogenous growth hormone can attenuate weight loss and preserve host body composition in tumor-bearing rats treated with doxorubicin without stimulating tumor growth. PMID- 8221689 TI - Biosynthetic basis of incompatible histo-blood group A antigen expression: anti-A transferase antibodies reactive with gastric cancer tissue of type O individuals. AB - The expression of incompatible A carbohydrate antigens in some adenocarcinomas may provide an explanation for the generally observed lower incidence of adenocarcinoma among types O and B versus type A individuals. The chemistry and genetic basis of incompatible A expression is largely unknown. Here, we have screened 31 cases of gastric tumors of phenotype O for the expression of blood group A gene-defined glycosyltransferase by immunohistology on frozen sections using newly developed monoclonal antibodies to the transferases. Three cases were positive, and transferase expression was confirmed by enzyme analysis of extracts from the specimens. Blood group A carbohydrate antigens were also identified immunohistologically in these three cases as well as in five other cases. Thin layer chromatography immunostaining analysis of glycolipid extracts from the three cases did not confirm the chemical presence of A antigen. The ABO genotype of all patients was found to be OO, showing that all carried O alleles with a structural defect at nucleotide position 261 leading to a shift in the reading frame. The data suggest that incompatible A antigen expression is a result of transferase expression derived from the ABO genes. PMID- 8221690 TI - Sezary T-cell-activating factor induces functional interleukin 2 receptors on T cells derived from patients with Sezary syndrome. AB - Sezary syndrome is the leukemic form of cutaneous T-cell lymphoma characterized by circulating neoplastic CD4+ T-cells. Although peripheral blood mononuclear cells from patients with Sezary syndrome have been shown to respond poorly to mitogens, we found that mitogen-activated peripheral blood mononuclear cells from four of five patients with Sezary syndrome produce a 28 kDa protein termed Sezary T-cell activating factor (SAF). SAF renders nonproliferating "resting" T-cells from leukemic patients or healthy donors responsive to interleukin 2 in the absence of a costimulator. We demonstrate that SAF induces functional, high affinity interleukin 2 receptors on T-cells from Sezary syndrome patients and provide evidence that SAF may be a novel cytokine. PMID- 8221691 TI - Macrophage nitric oxide synthesis delays progression of ultraviolet light-induced murine skin cancers. AB - The role of macrophages in the host immune response against cancers remains uncertain. Since nitric oxide synthesis represents a significant macrophage antitumor mechanism in vitro, we evaluated whether NO was synthesized during the immune response to growing murine skin cancers. NO synthesis was readily detectable in enzymatically dissociated tumors (RD-995 and LR-298) and was inhibited by N omega-monomethyl-L-arginine (MLA) and by macrophage depletion. Nitrosylation of iron-sulfur and heme complexes was observed in these tumors using electron paramagnetic resonance spectroscopy. NO production in the presence of increasing concentrations of MLA correlated inversely with tumor cell proliferation in vitro. To elucidate the role of NO during in vivo tumor progression, tumor-bearing mice were treated with continuous infusions of the nitric oxide synthase inhibitor MLA. MLA-treated mice demonstrated increased growth and delayed rejection of the highly antigenic UV radiation-induced regressor tumor LR-298. These experiments demonstrate that macrophage-derived NO synthesis can contribute to the antitumor immune response in vivo. PMID- 8221692 TI - Increased expression of high mobility group protein I(Y) in high grade prostatic cancer determined by in situ hybridization. AB - In a previous study using the Dunning rat prostate cancer model, we found high mobility group protein I-(Y) [HMG-I(Y)] to be overexpressed in metastatic tumor lines when compared to nonmetastatic lines. Hence, overexpression of this 12-kDa non-histone chromosomal protein may be associated with tumor progression. Firstly, by Northern analysis we showed that HMG-I(Y) expression increases in high grade prostate tumors. These studies, however, required fresh material, and clinical follow-up was limited. To overcome this problem paraffin-embedded material must be made amenable for determination of HMG-I(Y) expression in retrospective studies. RNA in situ hybridization enables the evaluation of mRNA levels in such material. We studied tumors from 71 patients with prostate cancer. The microscopic analysis of each sample included: (a) hybridization on sections with sense HMG-I(Y) and (b) 28S rRNA probes (nonspecific signal); (c) hybridization with antisense 28S rRNA (RNA preservation); (d) hybridization with an antisense HMG-I(Y) probe [quantification of HMG-I(Y) mRNA in the expressing areas]. Data were quantified using an image analysis system. High expression of HMG-I(Y) was observed in regions with high Gleason grade (4 and 5); whereas in lesions of Gleason grade 3, both weak and no expression was observed. In areas of grade 1 and 2, as well as in normal glands, low or no expression was found. We conclude that HMG-I(Y) expression assessed by RNA in situ hybridization is related to tumor differentiation in prostate cancer. These findings indicate that HMG-I(Y) expression may be a marker in prostate cancer diagnosis, and the possible clinical implication of expression of this gene in malignancy is discussed in this report. PMID- 8221693 TI - Production of transgenic mice expressing the Ki-ras oncogene under the control of a thyroglobulin promoter. AB - Transgenic mice have been generated bearing three fusion genes consisting of: (a) a 900-base pair rat thyroglobulin promoter followed by a gene coding for a chloramphenicol acetyl transferase activity; (b) the same promoter followed by the complementary DNA of the human activated Ki-ras oncogene; (c) a 2000-base pair rat thyroglobulin promoter followed by the complementary DNA of the human activated Ki-ras. We have shown that the 900-base pair rat thyroglobulin promoter is able to direct the expression of the reporter gene specifically in the thyroid gland of transgenic mice. The mice bearing the two Ki-ras constructs, which express the transgene in thyroid glands, show thyroid abnormalities, although at very low incidence. These lesions appear after a long latency and with a benign aspect, thus suggest that, in agreement with literature data on naturally occurring human thyroid tumors, the action of an activated ras gene is not sufficient to attain a complete malignant conversion of thyroid glands in vivo. However, ras expression in thyroid follicular cells represents a favorable ground for tumor development, as shown by the fact that goitrogen stimulation experiments increase the occurrence of tumors. PMID- 8221694 TI - Microvascular mechanisms of change in tumor blood flow due to angiotensin II, epinephrine, and methoxamine: a functional morphometric study. AB - To elucidate the microvascular mechanisms of change in tumor blood flow elicited by vasopressors, a functional morphometric study of the s.c. microcirculation within a rat transparent chamber was performed. Arteriolar vessels were classified centripetally (a2-a5) according to Strahler's method. Arteriolar pressure in each segment both under normotension and under hypertension induced by angiotensin II, epinephrine, or methoxamine was measured using a microocclusion technique. Vasoconstriction was estimated by changes in vessel diameters. In addition, tissue blood flow of the subcutis and s.c. tumor (LY80, a variant of Yoshida sarcoma) under the same conditions was measured with the hydrogen clearance method. By comparing the sites of the greatest pressure drop and the vasoconstriction induced by each vasopressor, we assessed the sites of vascular resistance (VR) which showed increases due to these vasopressors. The greatest VR increase elicited by angiotensin II occurred across a2 vessels. On the other hand, the sites of VR increase due to epinephrine were in a3 vessels and larger vessels upstream from a3 arterioles. The VR increase induced by methoxamine was much smaller than that induced by epinephrine. We conclude that the fact that the sites of increased VR differ with each vasopressor is the primary reason that various vasopressors have been found to produce different changes in tumor blood flow. PMID- 8221695 TI - Coamplification of the CDK4 gene with MDM2 and GLI in human sarcomas. AB - The 34-kilodalton cyclin-dependent kinase, p34cdk4, is a major catalytic subunit of mammalian D-type cyclins, which act during the G1 phase of the cell cycle to enforce the decision of cells to enter S phase. A murine complementary DNA clone was used to clone the cognate human CDK4 gene, which was localized to human chromosome 12, band q13, by fluorescence in situ hybridization. Because this chromosomal band contains the GLI and MDM2 genes, which are frequently amplified in human sarcomas, we analyzed CDK4 copy number and expression in a panel of sarcoma cell lines. An osteosarcoma cell line, OsACL, manifested a 25-fold increased copy number of CDK4, amplified concordantly with both GLI and MDM2, whereas a rhabdomyosarcoma cell line, SJRH30, was found to have an amplicon that included CDK4 and GLI but not MDM2. CDK4 mRNA and protein were overexpressed in both cell lines, and nucleotide sequencing analysis indicated that the gene had not sustained mutations. These observations provide the first evidence for amplification of a gene encoding a cell division cycle protein kinase, complement recent data indicating that genes encoding D-type cyclins are targets of chromosomal rearrangement and gene amplification in tumor cells, and suggest that CDK4 amplification might contribute to oncogenesis. PMID- 8221696 TI - Chromosome changes characterizing in vitro response to radiation in human squamous cell carcinoma lines. AB - We have analyzed the karyotypes of a series of 16 cell lines derived from human squamous cell carcinomas of the head and neck. When the cell lines were grouped according to their in vitro response to radiation, it was observed that some recurrent chromosomal changes occurred with differing frequencies between groups. Radiation resistance was associated with clusters of breakpoints in bands 1p22, 3p21, and 8p11.2 and deletion of distal 14q, while relative radiation sensitivity was associated with a high frequency of breakpoints in 11q13 and duplication of distal 14q. The regions identified point to the possible locations of genes involved in the response to radiation and could provide a series of markers with which to predict response to radiation therapy. PMID- 8221697 TI - Differential modulation of protein kinase C isoforms in erythroleukemia during induced differentiation. AB - Induction of erythroid differentiation of murine erythroleukemia cells (MELC) by exposure to hexamethylene bisacetamide (HMBA) involves the modulation of protein kinase C (PKC) activity. Using immuno- and Northern blot techniques, we have demonstrated that MELC express a pattern of PKC isoforms which includes PKC alpha, PKC delta, PKC epsilon, PKC zeta, and PKC eta. We show that MELC resistant to induction by HMBA express significantly less of the nPKC isoform, PKC delta, and slightly less PKC epsilon. Recovery of HMBA sensitivity is associated with reexpression of PKC delta protein. Upon exposure to HMBA, there is a fall in cytosolic PKC delta and PKC epsilon accompanied by a transient increase in membrane-associated forms of these PKC isoforms. HMBA-resistant MELC fail to display this isoform-specific translocation of PKC. Induction of differentiation is accompanied, over the next 24 h of exposure to HMBA, by a progressive fall in cellular PKC activity, associated with a progressive fall in the cellular content of PKC delta, PKC epsilon, and PKC zeta. These studies suggest that PKC delta, and possibly PKC epsilon and PKC zeta as well, play a role in the pathway of HMBA mediated terminal cell differentiation of MELC. PMID- 8221698 TI - Nicotine stimulates a serotonergic autocrine loop in human small-cell lung carcinoma. AB - Small-cell lung carcinoma cells express different plasma membrane nicotinic acetylcholine receptor subtypes. We have now found that interacting with these receptors (-)-nicotine induces a dose-dependent and stereoselective release of [3H]serotonin which is dependent on external calcium and blocked by the specific ganglionic nicotinic antagonist mecamylamine. With the same potency (-)-nicotine stimulates tumor cell proliferation, an effect also blocked by mecamylamine. Serotonin itself stimulates cell proliferation in a dose-dependent manner, an effect blocked by the selective serotonergic receptor antagonists methiotepine and metergoline. These data suggest that nicotine might affect proliferation of small-cell lung carcinoma cells by inducing the release of hormones (such as serotonin) with autocrine capabilities and place both the nicotinic and the serotonergic receptors at key positions in the biological and, possibly, pharmacological approach to this human lung cancer. PMID- 8221699 TI - Systemic expression of HIV-1 tat gene in transgenic mice induces endothelial proliferation and tumors of different histotypes. AB - The human immunodeficiency virus tat protein, a transactivator of viral and cellular genes, is suspected to be involved in the pathogenesis of acquired immunodeficiency syndrome-associated tumors. We report that transgenic mice carrying a recombinant DNA containing BK virus early region and the human immunodeficiency virus tat gene develop skin leiomyosarcomas, squamous cell papillomas and carcinomas, adenocarcinomas of skin adnexa, glands, and B-cell lymphomas. Although the incidence of hepatocellular carcinoma is low, most animals show a liver cell dysplasia of variable degree. These mice are also affected by skin lesions resembling the early stages of Kaposi's sarcoma. The transgene was detected intact in all the organs of transgenic mice, generally as multiple tandemly integrated copies. BK virus early region and tat were expressed in essentially all tissues and organs of BK virus/tat transgenic mice. This transgenic mouse model is representative of the systemic involvement of tat in human immunodeficiency virus natural infection and may be applied to investigate the role of tat in malignancies associated to acquired immunodeficiency syndrome, to study Kaposi's sarcoma pathogenesis and cell of origin, to characterize preneoplastic conditions established by tat in the skin and liver, and to assess in vivo the efficacy of antiangiogenic and anti-tat-specific drugs. PMID- 8221700 TI - New treatment strategies for bacterial meningitis. AB - Over the last several years, the therapeutic approach to bacterial meningitis has been changing. The neurological complications of bacterial meningitis result from an inflammatory process that is initiated by the lysis of bacteria in the subarachnoid space by antibiotics; to alter the pathophysiologic events initiated by this lysis, adjunctive therapy to antimicrobial therapy has been investigated in experimental and clinical trials. This article reviews our present understanding of bacterial meningitis, the efficacy of dexamethasone in treatment, and the potential use of leukocyte monoclonal antibodies, pentoxifylline, nonsteroidal antiinflammatory agents, and anticytokine antibodies. PMID- 8221701 TI - Neurobiological mechanisms involved in antidepressant therapies. AB - Selective serotonin reuptake inhibitors (SSRIs) are effective in alleviating the symptoms of depression. However, clinical improvement is only obtained after several weeks of treatment. SSRIs, when administered acutely to animals, have little effect on synaptic levels of serotonin. This suggests the existence of one or more regulatory mechanisms controlling serotonergic neurotransmission. The firing rate of dorsal raphe serotonergic neurons is under the control of somatodendritic 5-hydroxytryptamine 1A (5-HT1A) autoreceptors, the release of serotonin from nerve terminals is under the control of 5-HT autoreceptors (5-HT1B subtype in rodents, 5-HT1D in other species), whereas the control of the activity of tryptophan hydroxylase, the rate-limiting enzyme of serotonin synthesis, is complex, involving 5-HT1A but possibly other 5-HT receptors including the 5 HT1B/D subtype. During prolonged administration with a SSRI, these three feedback systems become desensitized and their regulatory effects on serotonergic neurotransmission are weakened or lost. This has the effect of allowing the synaptic levels of serotonin to rise with a consequently increased stimulation of one or more types of postsynaptic 5-HT receptor. Thus, it is only after prolonged administration that the pharmacological activity of SSRI is fully expressed in terms of synaptic serotonin levels. This may explain the latency of antidepressant action seen with these drugs in humans. Various other classes of antidepressant therapies (tricyclic antidepressants and monoamine oxidase inhibitor drugs, electroconvulsive therapy) have long-term effects on one or more of the feedback mechanisms such that an increase in synaptic concentrations of serotonin may be a common mechanism of many antidepressant therapies. PMID- 8221702 TI - Molecular cellular and behavioral aspects of peripheral-type benzodiazepine receptors. AB - Peripheral-type benzodiazepine receptors (PBR) are prominent in peripheral organs, whereas in the brain, they are sparse and located mainly in glial cells. The PBR bind with high affinity the ligands Ro 5-4864 (4'-chlorodiazepam) and PK 11195 (an isoquinoline carboxamide derivative), but not clonazepam, which binds with high affinity to central-type benzodiazepine receptors (CBR). Subcellularly, PBR are predominantly localized on the outer mitochondrial membrane. It appears that the PBR are composed of three subunits: an 18-kDa subunit that binds isoquinoline carboxamide derivatives; a 30-kDa subunit that binds benzodiazepines; and a 32-kDa subunit labeled by the benzodiazepine [3H]AHN 086, the voltage-dependent anion channel. Recently, complementary DNA (cDNA) encoding for rat and human PBR was isolated and sequenced. The PBR gene is located in the q13.3 region of the long arm of human chromosome 22. The PBR play a major role in steroidogenesis, controlling cholesterol mitochondrial transport. Diazepam binding inhibitor and its processing products, as well as porphyrins, have been suggested as putative endogenous ligands for these receptors. The PBR ligands have been shown to control cell proliferation and differentiation, and the binding capacity for these ligands is enhanced in some malignant tumors. Stress has been demonstrated to affect PBR bidirectionally. Acute stress is associated with increased PBR density, whereas chronic stress down-regulates PBR. PMID- 8221703 TI - Thyrotropin-releasing hormone produces different hemodynamic effects in vegetative and brain-dead patients. AB - To define a mechanism for the pressor effects of thyrotropin-releasing hormone (TRH), we evaluated changes in mean blood pressure (MBP) when a synthetic form of TRH (0.1 mg/kg, i.v.) was injected into two types of comatose patients: vegetative and brain dead. The patients in the vegetative group (n = 7, age 58 +/ 6) retained spontaneous respiration and brainstem function, whereas the brain dead (BD) patients (n = 7, age 68 +/- 4) lacked these functions. In the vegetative group, TRH caused significant increases in MBP (from 91 +/- 8 mm Hg to 110 +/- 10 mm Hg) at 2 min after the injection [p < 0.05, analysis of variance (ANOVA) with a Scheffe F-test]. In contrast, five of the seven BD patient showed no alterations in the measured parameter in response to the TRH injection. However, the remaining two BD patients, who had spinal reflexes, exhibited an elevation in MBP. In such BD patients, baroreceptor reflex function was virtually absent, suggesting that the blood pressure regulation mediating through the baroreceptor reflex system might be abolished. These results indicate that in comatose patients, the hemodynamic effects of TRH may differ depending on impairments in the central nervous system; the results support previous reports indicating a mediation of the central sympathetic nervous system in the development of pressor effects of TRH. Furthermore, because brain-dead patients with spinal reflexes showed hypertensive responses to TRH, there is a possibility that these responses may have resulted from an activation of TRH receptors in the spinal cord. PMID- 8221704 TI - Short-term memory in Parkinson's disease after withdrawal of long-term anticholinergic therapy. AB - We assessed the short-term memory (STM) using the Visual Paired Associates and the Verbal Paired Associates test, and the motor status at the end of 7.8 +/- 3.7 years of anticholinergic therapy in 22 nondemented patients with Parkinson's disease (PD). Eighteen patients managed to stop anticholinergic therapy. During anticholinergic therapy, STM in PD is significantly worse than in normal controls (CS). Two months after the withdrawal of anticholinergic drugs, the STM retest gain is significantly greater in PD than in CS, and at that time STM in PD did equal CS levels. After withdrawal of anticholinergic therapy, thirteen patients noticed a deterioration of motor function; the dose of levodopa had to be increased in seven patients. We conclude that in nondemented PD patients, long term anticholinergic therapy probably does not result in irreversible damage to STM, and withdrawal of long-term anticholinergic therapy is feasible in the majority of PD patients. PMID- 8221705 TI - Effect of D-penicillamine on pharmacokinetics of levodopa in Parkinson's disease. AB - A 42-year-old man had suffered from Parkinson's disease for 5 years. Levodopa was effective, but the wearing-off phenomena were severe. Because of relatively low levels of serum copper and ceruloplasmin, D-penicillamine was administered. D penicillamine increased plasma levodopa concentrations, thereby improving his parkinsonian symptoms. We propose that D-penicillamine facilitates levodopa absorption and, hence, the efficacy of the antiparkinsonian drug. PMID- 8221706 TI - Parkinsonian syndrome in childhood after sodium valproate administration. AB - Among the side effects attributed to sodium valproate administration, the production of a parkinsonian syndrome is very uncommon, particularly in children. We report a 12-year-old girl with secondary epilepsy; 7 days after the initiation of valproate therapy she developed parkinsonism that disappeared completely when valproate was replaced by carbamazepine. We discuss the possible role of alterations in GABAergic neurotransmission in the extrapyramidal syndrome that developed. PMID- 8221707 TI - Effect of nicardipine on essential tremor: brief report. AB - The effects of nicardipine, a calcium channel blocker (CCB), were investigated in 11 patients with essential tremor (ET). In a placebo controlled study, a single oral dose of 30 mg of nicardipine was administered, followed by 1 month of sustained treatment (60 mg/day). Tremor was assessed by accelerometric recording. A single oral dose of nicardipine reduced the tremor amplitude respect baseline (p = 0.003) and placebo (p = 0.008). After 1 month of chronic treatment, nicardipine still reduced the tremor amplitude (30.78% +/- 17.13 SE from baseline), but failed to sustain the initial statistical improvement. A single oral dose of nicardipine is effective in reducing ET. PMID- 8221708 TI - Flunarizine in essential tremor. AB - We studied the effects of flunarizine (Fz) in 10 patients with moderate to severe essential tremor. Tremor was evaluated after 6 weeks of treatment using patient and physician assessment as well as blinded video analysis. Only one patient had mild subjective transient improvement and three experienced worsening of tremor. No patient elected to remain on Fz. We conclude that Fz is ineffective for moderate to severe essential tremor and may actually worsen the symptoms in some patients. PMID- 8221709 TI - Successful treatment of spinal sarcoidosis by high-dose intravenous methylprednisolone. AB - A man with known pulmonary sarcoidosis presented with paraplegia and a rod-shaped increase in T2 signal intensity in his cervical and upper thoracic spine. Initial treatment efforts using oral doses of 40 mg of methyl-prednisolone were futile, but intravenous bolus therapy (500 mg of methyl-prednisolone daily for 1 week) led to long-lasting improvement of his neurological status and to normalization of MRI findings. PMID- 8221710 TI - Intravenous midazolam suppression of complex partial status refractory to intravenous phenytoin and diazepam. PMID- 8221711 TI - Pneumocystis carinii pneumonia in cancer patients. AB - Concern has been arisen about the recently reported increasing incidence of PCP in patients with cancer and the potential transmissibility of this infection. Whether or not there is an increase in the incidence of P. carinii infections, PCP should be considered in the differential diagnosis of pulmonary infiltrates in bone marrow transplant recipients, in patients with hematologic neoplasms and in patients with primary or metastatic brain neoplasms. Intensity of immunosuppression plays a crucial role, especially long-term (> 2 months) corticosteroid treatment. PCP is usually manifested clinically during augmentation or during tapering of corticosteroid dose. Thus, if the chest radiograph of a high-risk patient shows diffuse infiltrates, bronchoscopy and bronchoalveolar lavage should be done immediately. Treatment options are the same as for the AIDS population, except that TMP-SMX is tolerated better in non-AIDS patients. The role of supportive care, including mechanical ventilation in such patients should not be underestimated. Oral therapy with dapsone-trimethoprim or with atovaquone, can be as effective as conventional therapy in mild disease, permitting treatment on an outpatient basis. PCP is often preventable and our understanding has improved about when prophylaxis should be initiated. In the future, the emergence of new technologies for diagnosis and of new agents for treatment and prophylaxis, will bring us closer to the goal of controlling this serious infection. PMID- 8221712 TI - Carboplatin-based chemotherapy for bladder cancer. AB - Urothelial cancer is common and the prognosis of patients with locally advanced tumors treated with conventional treatment is poor. In this trial we examined responses among 72 urothelial cancer patients referred for treatment with MVMJ (methotrexate/vinblastine/mitoxantrone/carboplatin) chemotherapy. Sixty-four evaluable patients, 37 with locally advanced and 27 with metastatic urothelial cancer, were treated. Twenty-nine (45%) of the 64 patients had a complete or partial response, 15 (23%) had stable disease, and 13 (20%) had disease progression. The median survival of the responding patients has not been reached (range, 114 to 1184+ days). Twelve patients (32%) with locally advanced disease had a complete response to treatment; their median survival has not been reached and ranges up to 1131+ days. Five patients (18%) with metastatic cancer had a complete response to treatment and their median survival was 497 days (range, 184 to 637+). There were seven deaths within the first treatment month. PMID- 8221713 TI - Long-term oral etoposide in metastatic breast cancer: clinical and pharmacokinetic results. AB - To evaluate the activity of long-term, single-agent oral etoposide against advanced breast cancer, this study employed etoposide 50 mg/day and 100 mg/day (given over 14 days) in previously treated and chemotherapy-naive patients with histologically confirmed, recurrent or metastatic breast cancer. Of 38 patients, 24 had had chemotherapy, 34 had prior radiotherapy, and 31 had received hormone therapy. Etoposide courses in both treatment groups were repeated every 4 weeks for at least two courses; delays were instituted when patients' total leukocyte nadir fell to or below 3.0 x 10(9)/l. Dose reductions were made in the 100-mg group (to 50 mg/day) if World Health Organization leukopenia grade 3 or higher was present. Plasma pharmacokinetic profiles were measured in selected patients to assess inter- and intrapatient variability in etoposide's oral bioavailability. No complete responses were achieved among the 36 patients evaluable for response, but eight patients had a partial response. Responses were more frequent at the 100-mg dose and in previously untreated patients (seven of eight partial responders had not had previous chemotherapy). Median duration of response was 16 weeks (range, 7 to 46). Myelosuppression (variable and unpredictable) and alopecia (universal) were the notable toxicities. Pharmacokinetic analyses of oral bioavailability revealed significant interpatient variability, but much less intrapatient variability when successive etoposide courses in individual patients were evaluated. Despite the relatively small number of patients in this study, the responses achieved by previously untreated patients suggest etoposide's value against breast cancer. Further trials should use pharmacokinetic studies to assess bioavailability as well as to help define 'target' etoposide doses, based on plasma etoposide levels, during treatment. PMID- 8221714 TI - Carboplatin in combination as first-line therapy in advanced breast cancer. AB - Carboplatin, a platinum analog with single-agent activity in previously untreated breast cancer, is characterized by comparatively less renal toxicity and emesis than cisplatin. We combined carboplatin at different dose levels [from 200 to 350 mg/m2 by intravenous (IV) infusion on day 1] with 5-fluorouracil (500 mg/m2 IV on days 1 and 8) and cyclophosphamide (500 mg/m2 IV on day 1), with all three drugs recycled every 28 days, to evaluate anti-tumor activity and toxicity of this novel combination [5-fluorouracil/carboplatin/cyclophosphamide (FCC)] in untreated locally advanced (LABC) or metastatic breast cancer (M+). Of 37 patients treated between March 1990 and August 1991 [LABC 25, M+ 8; World Health Organization (WHO) performance status, 0-1; median number of treatment cycles, 5; median follow-up, 20 months], 33 are evaluable for response and toxicity. The overall complete plus partial remission rate was 57% (LABC 68%, M+ 25%). The median duration of response was 19+ months. The cumulative carboplatin dose ranged from 800 to 2350 mg/m2 (median, 1450 mg/m2). In this series, no correlation was observed between the carboplatin dose level and response rate or toxicity. Leukopenia and thrombocytopenia represented the most frequent toxicities. WHO grades 3 and 4 neutropenia were documented in 34% and 8% of patients, respectively. Thrombocytopenia below 50 x 10(9)/l was observed in 8%. No renal toxicity was observed, and moderate emesis occurred in 67% of patients. These results indicate that FCC is an active and relatively safe combination for the treatment of advanced breast cancer in patients not previously treated with chemotherapy. PMID- 8221715 TI - Chronic etoposide administration: overview of clinical experience. AB - Etoposide is an important anti-neoplastic drug, but the best dose and schedule for its administration remain unknown. The schedule-dependency of etoposide for small cell lung cancer (SCLC) is now proven and it is probably true for other sensitive neoplasms as well (e.g., lymphoma and germ cell tumors). Recent data suggest that an extended schedule of administration (i.e., 14 to 21 days) may be more effective than the standard 3- to 5-day schedule. Several phase II studies with etoposide have demonstrated its activity against several neoplasms. Some have assessed the relationship of etoposide plasma levels to dose, schedule, and tumor responsiveness. Preliminary data suggest that high peak plasma levels (i.e., > 5 to 10 micrograms/ml) are associated with more severe myelosuppression than lower peak plasma levels (i.e., 1 to 3 micrograms/ml). Response and survival rates in previously untreated SCLC patients given low daily doses of etoposide for 14 to 21 days are at least comparable with results achieved with standard etoposide doses given for 3 to 5 days, and the extended etoposide schedule is less toxic than the 3- to 5-day schedule. Preliminary data from several studies suggest that administering low, daily etoposide doses over a prolonged schedule is a superior method of administration. Other studies, including randomized comparisons, are necessary to confirm these observations. PMID- 8221716 TI - Preliminary results of a phase II trial of chronic oral etoposide in breast cancer. AB - Twenty-seven women with metastatic breast cancer (at least one site of measurable disease) entered a phase II study of chronic oral etoposide (50 mg/m2/day x 21 days, given every 4 weeks). To date, 23 patients are evaluable for response and toxicity. All patients had received prior chemotherapy (adjuvant therapy, one patient; adjuvant plus chemotherapy for metastases, six patients; chemotherapy for metastases, 16 patients). Thirteen patients had previously received anthracyclines, and 10 had also received prior hormonal therapy. Of the 23 evaluable patients, one obtained a complete response and six achieved partial responses (objective response rate 30.4%, 95% confidence interval, 13 to 53%). Responses were seen in lymph nodes (three of eight sites), skin and soft tissue (five of seven), lung (two of six), lytic lesions of the bone (one of three), and liver (1 of 12). The median duration of responses was 6 months (range, 1+ to 8). The main toxic side-effects were leukopenia (74% of patients), thrombocytopenia (22%), and anemia (69.5%). Myelosuppression in four patients (17%) necessitated a 25% dose reduction. Other toxicities included alopecia (83%), mucositis (52%), and emesis (35%). Chronic oral etoposide appears to be an active regimen in metastatic breast cancer patients previously exposed to chemotherapy. PMID- 8221717 TI - Carboplatin, etoposide, and radiotherapy, followed by surgery, for the treatment of marginally resectable non-small cell lung cancer. AB - The present study was undertaken in order to determine the feasibility and efficacy of induction chemotherapy with carboplatin and etoposide, followed by weekly carboplatin and full-course radiotherapy as pre-operative therapy for marginally resectable non-small cell lung cancer (NSCLC). Twenty-eight patients with good Eastern Cooperative Oncology Group (ECOG) performance status ratings and stage IIIA NSCLC received induction chemotherapy with carboplatin (dose computed with the Egorin formula, days 1 and 29) and etoposide (100 mg/m2/day, days 1 through 3 and 29 through 31). This was followed by 100 mg/m2 weekly carboplatin given over 6 weeks, concurrently with 60 Gy radiotherapy. Patients with either responsive or stable disease underwent thoracotomy 4 weeks after the completion of combined-modality therapy. All 28 patients received the first chemotherapy cycle (average carboplatin dose, 407 mg/m2; range, 195 to 586 mg/m2). World Health Organization (WHO) grade 3/4 neutropenia and thrombocytopenia were observed in 53 and 34% of patients, respectively. There were three febrile neutropenic episodes, but no septic deaths. Five patients (18%) required dose reductions prior to the second chemotherapy cycle, but the dose intensity of carboplatin was maintained (average dose, 390 mg/m2; range, 195 to 586 mg/m2). In all, 82% of patients received full-dose radiotherapy, and 73% received at least five of six planned concurrent weekly carboplatin doses. Carboplatin doses were most frequently delayed for thrombocytopenia and/or leukopenia. Carboplatin did not increase the incidence of radiation-induced esophagitis. Only three patients required interruption of radiotherapy, for esophagitis (two patients) and persistent thrombocytopenia (one patient). The response rate to pre-operative therapy was 64%. In this study, we demonstrated the ability to deliver escalated doses of carboplatin with standard-dose etoposide as induction chemotherapy with reasonable myelotoxicity. The combined modality therapy was well tolerated, and the addition of weekly carboplatin did not result in increased radiation-related toxicity. This neoadjuvant regimen is active in the treatment of locally advanced NSCLC, and compares favorably to other cisplatin-based regimens. PMID- 8221718 TI - A convenient synthesis of N-acetyllactosamine derivatives from lactal. AB - In a thermal inverse-type hetero-Diels-Alder reaction of O-silyl-protected lactal 1 and bis(2,2,2-trichloroethyl) azodicarboxylate (2), the dihydrooxadiazine derivative 3 was obtained in a very high yield; transesterification with benzyl alcohol furnished the corresponding derivative 4. Treatment of 3 with methanol in the presence of BF3.OEt2 afforded the methyl lactoside derivative 5 which, after transesterification with benzyl alcohol, then hydrogenolytic debenzylation and concomitant NN-cleavage with Raney nickel, and N-acetylation, furnished methyl O (2,4,6-tri-O-tert-butyldimethylsilyl-beta-D-galactopyranosyl)-(1-->4)- 2 acetamido-3,6-di-O-tert-butyldimethylsilyl-2-deoxy-beta-D-glucopyra nos ide (7) in high yield. Desilylation of 4, then O-acetylation, methyl glycoside formation with methanol-BF3.OEt2, hydrogenolytic debenzylation, and NN-cleavage with Raney nickel, and N-acylation afforded methyl O-(2,3,4,6-tetra-O-acetyl-beta-D galactopyranosyl)-(1-->4)-2-acetamido-3 ,6-di-O-acetyl-2-deoxy-beta-D glucopyranoside (10). PMID- 8221719 TI - Characterization of inhibitory activities and binding mode of synthetic 6' modified methyl N-acetyl-beta-lactosaminide toward rat liver CMP-D-Neu5Ac: D galactoside-(2-->6)-alpha-D-sialyltransferase. AB - 6'-Deoxy (12), 6'-thio (13), and 6'-O-tetrahydropyranosyl (14) analogues of methyl N-acetyl-beta-lactosaminide (3), were synthesized from lactose. NOE experiments proved that they adopt the same conformation as that of 3. Inhibition studies using these synthetic analogues, including the disulfide dimer 15, toward (2-->6)-alpha-sialytransferase (EC 2.4.99.1) revealed that the 6'-deoxy analogue 12 had remarkable inhibitory activity as the first acceptor-analogue inhibitor for this enzyme. It is noteworthy that the disulfide 15 also behaves as an inhibitor. The results indicated that chemical modification at the 6'-position of 3 did not cause much decrease in the binding affinity to the sialytransferase. Further, a novel possibility that the acceptor and the acceptor-analogue inhibitor can bind simultaneously to the sialytransferase was proposed based on the inhibition studies with 12 and CMP. PMID- 8221720 TI - Determination of the pattern of methyl esterification in pectin. Distribution of contiguous nonesterified residues. AB - A method is described for determining the distribution of contiguous nonesterified galacturonic acid residues within pectins. First, the esterified galacturonic acids are converted to galactose by reduction with sodium borohydride, then the glycosidic linkages of the resulting galactose residues are cleaved selectively by liquid HF solvolysis. Separation and quantitation of the resulting galacturonic acid containing oligomers reveals the proportion of each stretch of contiguous nonesterified galacturonic acid residues in the original pectin. The distribution of nonesterified GalA in a pectin fraction obtained from cotton suspension culture cell walls with approximately 50% esterification appears to be far from random. PMID- 8221721 TI - "Fast moving" and "slow moving" heparins, dermatan sulfate, and chondroitin sulfate: qualitative and quantitative analysis by agarose-gel electrophoresis. AB - Heparin from beef intestinal mucosa, dermatan sulfate from beef intestinal mucosa, and chondroitin sulfate from bovine trachea were extracted and purified, and their structures and physico-chemical properties were evaluated by different techniques (disaccharide patterns by specific enzymatic cleavage, relative molecular mass by high-performance size-exclusion chromatography, sulfate-to carboxyl ratio by potentiometric determination). Heparin was fractionated into "slow moving" and "fast moving" fractions by selective precipitation as the barium salt at different temperatures. The "fast moving" and "slow moving" components of heparin, dermatan sulfate, and chondroitin sulfate were utilized to run calibration curves in agarose-gel electrophoresis. Mixtures containing different amounts of these glycosaminoglycans were made and separated by agarose gel electrophoresis, and these were analyzed quantitatively. For analysis of relative amounts, the area of each individual component of mixtures, obtained by photodensitometric readings, was divided by the sum of the areas of all glycosaminoglycans and expressed as a percentage. For analysis of absolute amounts, the area under the curve for each component of mixtures was fitted to specific calibration curves, and the amount of each glycosaminoglycan was calculated in micrograms. The quantitative procedure performed by analysing absolute amounts was used to obtain an accurate quantitative evaluation of each component in mixtures of glycosaminoglycans utilized for pharmaceutical purposes. A sensitive method was developed for the evaluation of very small amounts (0.2% w/w) of possible glycosaminoglycans as contaminants in preparations of a single species of glycosaminoglycan. This technique requires specific enzymatic degradation by bacterial lyases, separation in agarose-gel electrophoresis, and quantitative analysis by photodensitometric analysis and specific calibration curves. PMID- 8221722 TI - Characterization of the oligosaccharide structures on bee venom phospholipase A2. AB - The N-linked oligosaccharide structures on bee venom phospholipase A2 were investigated. The oligosaccharides on purified phospholipase A2 were released by hydrazinolysis and labeled in vitro by reduction with NaB3H4. Following purification, the labeled oligosaccharides were characterized by size exclusion chromatography in combination with digestion with specific glycosidases. Linkage positions were determined by methylation analysis. Four types of structures were identified on the molecule, all of which were of truncated high-mannose type and none of which contained any alpha-(1-->2)-linked mannose residues. The majority of the structures were Man3 oligosaccharides with (43%) or without (38%) a fucose residue linked alpha-(1-->6) to the reducing N-acetylglucosamine. The remaining 19% of the oligosaccharides on the molecule were identified as a Man5 oligosaccharide without core fucose (9.6%) and a core-fucosylated Man4 structure (9.2%). PMID- 8221723 TI - Molar-response factors for the quantitative analysis of fully methylated methyl 2 acetamido-2-deoxyhexopyranosides by gas-liquid chromatography with flame ionization detection. PMID- 8221724 TI - Synthesis of the terminal trisaccharide unit of the lipo-oligosaccharide from Mycobacterium linda. PMID- 8221725 TI - Synthesis of branched-chain sugars with methiniminium salts. PMID- 8221726 TI - Syntheses of the octyl and tetradecyl glycosides of 3,6-di-O-alpha-D mannopyranosyl-alpha-D-mannopyranose and of 3,4-di-O-alpha-D-mannopyranosyl-alpha D-mannopyranose. A new way for 2,4-di-O-protection of mannopyranosides. PMID- 8221727 TI - Structure of glycosylamines and diglycosylamines in the arabinose, mannose, and rhamnose series. PMID- 8221728 TI - An imino-linked carba-disaccharide alpha-D-mannosidase inhibitor. PMID- 8221729 TI - A cellobiose phosphorylase from Cellvibrio gilvus recognizes only the beta-D-form of 5a-carba-glucopyranose. PMID- 8221730 TI - The tetrasaccharide nystose trihydrate: crystal structure analysis and hydrogen bonding. AB - The crystal structure of nystose trihydrate, O-beta-D-fructofuranosyl-(2-->1)-O beta-D-fructofuranosyl-(2-->1)-beta-D -fructofuranosyl alpha-D-glucopyranoside trihydrate, C24H42O21.3H2O, has been determined using Cu K alpha X-ray data at 121 K. The space group is P2(1)2(1)2(1), with 4 molecules in a unit cell of dimensions a = 10.155(1), b = 13.506(1), and c = 23.278(2) A. The structure was refined to R = 0.052 and RW = 0.048 for 2734 observed structure amplitudes. The alpha-D-glucopyranose unit of the molecule has the normal 4C1 chair conformation and the three fructofuranose units have twist conformations lying between E3 and 4T5. One of the three water molecules is distributed over two sites: W-3 with occupancy 0.80 and W-3' with occupancy 0.20. All the hydrogen atoms were located on the difference synthesis with the exception of those attached to the low occupancy water site. All hydroxyls, two of the three linkage oxygen atoms, and the water molecules are involved in a complex three-dimensional network which can be decomposed into a series of infinite chains intersecting at the water molecules to form homo- and hetero-dromic cycles. PMID- 8221731 TI - Computer modeling of the tetrasaccharide nystose. AB - Nystose, O-beta-D-fructofuranosyl-(2-->1)-O-beta-D-fructofuranosyl-(2-->1)- beta D-fructofuranosyl alpha-D-glucopyranoside, was modeled with the rigid-residue program PFOS and with MM3. The three furanose rings of crystalline nystose trihydrate all are within the 1 kcal.mol-1 contour on an MM3 energy surface that was calculated with a dielectric constant of 4. The calculations showed that in the extended conformation found in the crystal, there was a small influence of the remainder of the residues on the conformation of any given disaccharide segment. Even accounting for intramolecular, interresidue forces, however, the central inulobiose and the sucrose linkages appear to be distorted. These discrepancies are apparently due to crystal packing forces and, for the sucrose linkage, miscalculation of the energy of an overlapping exoanomeric effect, as indicated by studies of a model miniature crystal. The different conformations of the central and terminal inulobiose linkages in the crystal show that conformations in higher oligosaccharides will not always correspond to the global minima on conformational maps for isolated disaccharides. PMID- 8221732 TI - The crystal structure and conformational analysis of substituted 2,7 dioxabicyclo[4,1,0]heptanes: 1,2-anhydro-3,4,6-tri-O-benzyl-beta-D-talopyranose. AB - The title compound, C28H27O5, is triclinic, space group P1 with unit cell dimensions a = 12.763(2), b = 11.130(2), c = 4.764(3) A, alpha = 73.78(3), beta = 82.89(3), gamma = 62.16(1) degrees, V = 574.8(4) A3, and Z = 1. The pyranose ring has an 4H5 conformation with some flattening at C-4. Molecular mechanisms calculations indicate that the 4H5 conformation of the pyranose ring in the title compound is the most stable conformation. PMID- 8221733 TI - The conformational analysis of methyl beta-xylobioside: effect of choice of potential functions. AB - In order to determine the effect of the choice of potential function used in the conformational analysis of a carbohydrate, the NMR spectrum of methyl beta xylobioside [beta-D-Xyl-(1-->4)-beta-D-Xyl-(1-->O)-Me] was interpreted using calculated J13C-H coupling constants and nuclear Overhauser effects for protons across the anomeric linkage. Conformational flexibility was described by calculating average phi and psi angles, and estimating their standard deviations. ECEPP2, ECEPP83, and HSEA potentials were used in the first series of calculations. The calculated coupling constants and nuclear Overhauser effects were averaged over the Boltzmann distribution of conformations of the disaccharide in which the entire phi, psi space was scanned in ten-degree steps while retaining fixed bond distances and angles in the remainder of the molecule. In the second series of calculations, MM2, MM2CARB, and PCILO parameters were used to calculate conformational energies. Conformational optimization was done. The effect of temperature and solvent on the calculated coupling constants was negligible. Calculated properties from conformations whose energies were based on the ECEPP parameters gave the best agreement with experiment. Exploration of the conformational space in breadth rather than on a detailed level of full optimization appears to be a preferable course of action. PMID- 8221734 TI - Crystal structure of 6-O-[(R)-2-hydroxypropyl]cyclomaltoheptaose and 6-O-[(S)-2 hydroxypropyl]cyclomaltoheptaose. AB - Crystal structures of 6-O-[(R)-2-hydroxypropyl]- and 6-O-[(S)-2-hydroxypropyl] cyclomaltoheptaose were determined by X-ray analysis. In both structures, the 2 hydroxypropyl group is inserted into the macrocyclic cavity of the next molecule related by the two-fold screw axis, and a helically extended polymeric structure is formed by repetition of the intermolecular inclusion. The hydroxyl group of the substituent group penetrates through the macrocyclic ring from the secondary hydroxyl side and is linked to an HO-6 group by a hydrogen bond. Comparison of intermolecular contacts of the substituent group indicates that the (S)-2 hydroxypropyl group is better fitted to the cavity than the (R)-2-hydroxy-propyl group. PMID- 8221735 TI - Microscopic protonation constants in tobramycin. An NMR and pH study with the aid of partially N-acetylated derivatives. AB - Three tetra-N-acetyl derivatives and one tri-N-acetyl derivative of tobramycin (1) have been prepared by partial N-acetylation. Comparison of the pKa values, determined by NMR chemical shift titrations and pH titration of the derivatives, with those of unprotected 1 suggests that protonation equilibria at any particular amino group in 1 are not likely to be influenced by those at other sites. pH-Dependent conformational changes in 1 were assessed on the basis of 1H and 13C chemical shift changes in the derivatives. PMID- 8221736 TI - [Changes in the physical characteristics of the lung can account for the improvement observed after ultrafiltration in patients with moderate heart failure]. AB - Ultrafiltration improves the clinical condition of patients with moderate congestive heart failure. Ultrafiltration (1895 +/- 120 ml of plasma water, rate 600 ml/h, veno-venous bypass) was performed in 30 patients with moderate congestive heart failure in stable clinical conditions. The effects of ultrafiltration were assessed through pulmonary function and cardiopulmonary exercise tests performed before and 30 days after the procedure. Vital capacity, forced expiratory volume (1 s) and maximal voluntary ventilation improved as improved exercise performance (increase in oxygen consumption at peak exercise and at anaerobic threshold). Tidal volume at anaerobic threshold and at peak exercise increased by 10% with no changes in respiratory rate. This suggests that ultrafiltration induces changes at the physical characteristics of the lungs probably related to changes in lung water content. PMID- 8221737 TI - [Anterograde and retrograde cardioplegia in myocardial revascularization. An intraoperative contrast echographic study]. AB - The aim of this study was to assess the distribution of antegrade and retrograde cardioplegia with intraoperative contrast echocardiography in patients undergoing coronary artery bypass grafting. Fifteen patients with chronic stable angina pectoris and severe coronary artery disease were studied. The severity of coronary artery disease was assessed at coronary angiography, using the Jeopardy Score System. The presence and the extent of collateral circulation was evaluated on the basis of preoperative coronary angiography and graded as: absent or poor; good or excellent. Coronary revascularization was carried out during extracorporeal circulation and myocardial protection was performed with antegrade (aortic root) and retrograde (right atrial) cardioplegia. The echo contrast agent was sterilely prepared 1 hour prior to surgery and consisted of a solution of sonicated 5% human albumin microbubbles. Two ml of sonicated albumin were injected along with antegrade cardioplegia and 4 ml with retrograde cardioplegia. The echocardiographic images were obtained with transesophageal echocardiography in the transgastric left ventricular short-axis view. Images were recorded on videotape for off-line planimetric measurement of percent myocardial opacification. Data were analyzed with the analysis of variance. Multiple comparisons were made with Student's paired t test and using Bonferroni's correction. Myocardial opacification was 58.9 +/- 12.9% during antegrade cardioplegia and 77.5 +/- 16.4% during retrograde cardioplegia (p = 0.003). This overall difference was mainly due to the impact of collateral circulation in the distribution of antegrade cardioplegia. Patients with absent or poor collateral circulation showed a lower degree of myocardial opacification than patients with good or excellent myocardial opacification (44.3 +/- 12.0% versus 64.2 +/- 8.6%; p < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221738 TI - [The clinical and echocardiographic parameters at rest and during mental arithmetic stress in preadolescent subjects with a hypertensive parent]. AB - Previous studies have showed that a genetic predisposition and/or a family history of essential hypertension may play a significant role in the development of cardiovascular changes in borderline hypertensive patients. Moreover, mental stress has been identified as contributing to the development of cardiovascular disease. The aim of this study was to compare the possible changes in blood pressure and Doppler echocardiographic parameters in resting and during mental arithmetic stress in 21 children with a hypertensive parent (Group I) and 21 age matched children with normotensive parents (Group II), similar for body mass index, alimentary and life habits, and not differing in resting blood pressure, heart rate and Doppler echocardiographic parameters. Mass index was slightly but not significantly high in Group I with respect to controls. During mental stress only in the children with a family history of hypertension there was a significant increase (p < 0.02) in systolic blood pressure. A positive relation between the increase in systolic blood pressure and end-diastolic thickness of interventricular septum and posterior wall was observed in the same group. In addition a close relation (p < 0.05) between mass index and end-systolic left ventricular stress at rest was evidenced, whereas a negative relation (p < 0.03) between the increase in systolic blood pressure on one side and the early peak diastolic filling velocity (Vmax E) and the ratio of early to atrial peak diastolic filling velocity (Vmax E/Vmax A), on the other side, were found.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221739 TI - [The prevalence, risk factors and clinical significance of atherosclerosis of the thoracic aorta: a transesophageal echocardiographic study]. AB - Available information on atherosclerosis of thoracic aorta in man is scanty and mostly derived from pathological or surgical series. Transesophageal echocardiography makes a clear definition of the entire thoracic aorta possible and enables large, population based studies. In order to define prevalence, risk factors and clinical implications of aortic atherosclerosis, the echocardiographic recordings of 220 patients suitable for both evaluation of thoracic aorta and risk factors analysis were reviewed. Transesophageal echocardiography has been performed because of valvular diseases (78), suspected aortic aneurysm or trauma (43), evaluation of valve prosthesis (39), previous cerebral or peripheral embolic events (22), infective endocarditis (14), cardiac mass lesions (12) or other indications (12). Age ranged from 5 to 81 years (55 +/ 15), male to female ratio was 0.99. Simple and complex atherosclerotic plaques were identified in 33% and 10% respectively. Complex atheromas were more frequent among patients with previous embolic episodes (6/22, 27% versus 17/198, 8.5%; p = 0.019). The prevalence of any type of atherosclerosis progressively increased from the fourth (8%) to the eighth (88%) decade of age. By univariate analysis age (p < 0.001), history of hypertension (p < 0.001), systolic (p < 0.001) and diastolic (p < 0.05) pressure, type II diabetes mellitus (p < 0.01), HDL cholesterol (p < 0.01), HDL/total cholesterol (p < 0.01) and uricaemia (p < 0.05) were associated with aortic atherosclerosis. Discriminant analysis identified 5 independent variables associated with the presence and the extent of atherosclerosis (Wilk's Lambda = 0.43): number of cigarettes per day, age, history of hypertension, systolic pressure and type II diabetes mellitus. This model provided a 63% correct classification rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221740 TI - [An ergometric cross-over evaluation of the anti-ischemic efficacy of amlodipine and gallopamil in stable angina of effort]. AB - In the setting of stable effort angina a single-blind, randomized, cross-over study to evaluate the effects of gallopamil (GAL) and amlodipine (AML) on exercise tolerance and ischemic ST depression was conducted. Fifteen outpatients, 12 males and 3 females, aged 40-65 years (57 +/- 9), with documented coronary atherosclerosis and reproducible ST-segment depression on 2 consecutive baseline exercise stress tests, completed the study, which consisted of 4 periods: 1 and 3 placebo, 2 and 4 at random GAL (50 mg tid) and AML (10 mg/daily). At the end of each period a multistage treadmill exercise stress test (Bruce protocol) was performed. Both drugs significantly (p = 0.0001) increased the ischemia time (IT) (0.1 mV ST depression) as compared to placebo, from 416 +/- 165 s to 635 +/- 161 s (GAL) and 607 +/- 152 s (AML) with significant difference (p = 0.2) between the 2 drugs, and reduced significantly (p = 0.001) the maximal ST depression from 0.25 +/- 0.09 mV to -0.11 +/- 0.08 mV (GAL) and -0.12 +/- 0.09 mV (AML). At the IT, the systolic blood pressure increased from 178 +/- 23 mmHg to 185 +/- 20 mmHg (GAL) and remained unchanged during AML treatment (178 +/- 15 mmHg); similarly, the heart rate increased from 126 +/- 22 b/min to 139 +/- 21 b/min (GAL) and 138 +/- 19 b/min (AML). In conclusion, both GAL and AML showed a good anti-ischemic effect (IT = +52.6% during GAL and +45.9% during AML), even if GAL proved to be significantly more effective than AML. PMID- 8221741 TI - [Captopril administration before thrombolysis in patients with an acute myocardial infarct: a pilot study of the short- and long-term effects]. AB - Several experimental studies, carried out on animals and on isolated heart, showed that captopril can reduce the post-ischemic reperfusion injury. Our study was aimed at checking the effects of captopril before thrombolysis in acute myocardial infarction (AMI) and included 204 patients, hospitalized within 4 hours from the onset of symptoms. Patients were randomly subdivided into 2 groups: the first group (105 patients, Group A: pretreatment) received 6.25 mg captopril orally about 15 min before iv administration of urokinase (2 million), the second group (99 patients, Group B: late-treatment) received captopril about 3 days after thrombolytic treatment. Captopril doses were later increased in both groups according to blood pressure. All patients were subdivided according to the localization of infarction. One hundred and thirty-seven patients showed anterior AMI (70 from Group A and 67 from Group B); 67 patients showed inferior AMI (35 from Group A and 32 from Group B). Ventricular hyperkinetic arrhythmias (VHA) due to reperfusion were evaluated during the first 2 hours. Ventricular hyperkinetic arrhythmias occurred in 11.4% of patients with anterior AMI in Group A versus 38.8% in Group B (p < 0.001). CK peak normalization time in the group with anterior AMI was achieved after 58 +/- 2 hours in Group A versus 72 +/- 2 hours in Group B (p < 0.01). Late arrhythmias, > Lown's class 2 was found to occur in 15.7% of patients with anterior AMI of Group A versus 32.8% in Group B (p < 0.05), at predischarge Holter test. One hundred and nineteen patients underwent a hemodynamic test about 3 weeks after AMI.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221742 TI - [Angiotensin receptors in the rat myocardium during pre- and postnatal development]. AB - Angiotensin II exerts positive inotropic and chronotropic effects on mammalian heart by binding to specific membrane receptors. Recently, 2 subtypes of angiotensin II receptors (AT1 and AT2) have been distinguished using the nonpeptide antagonists losartan and PD123177. Because angiotensin II has been reported to have growth potentiating effects in several tissues, we examined angiotensin II receptors in fetal (embryonic day 16 and 19), neonatal (1, 2, 3 and 10 days), and adult (10 and 16 weeks) rats. We performed an 125I-[Sar1,Ile8] angiotensin II in situ binding assay on tissue sections obtained from Sprague Dawley rats. Binding specificity was verified by competition with unlabeled [Sar1]-angiotensin II. Distribution of AT1 and AT2 receptors was determined by competition with losartan and PD123177, respectively, and the density of receptors quantitated by emulsion autoradiography. Angiotensin II receptors were widely distributed throughout the heart, with each receptor subtype accounting for approximately 50% of the specific binding. Binding density was comparable in right and left ventricles, and interventricular septum. Throughout cardiac development a significant increase (p < 0.005) in the density of both receptor subtypes was found immediately after birth, reaching a maximum on day 2, and decreasing toward prenatal values thereafter. No variation in the proportion of the 2 receptor subtypes was observed during development. Thus, in rat heart, AT1 and AT2 receptors are equally distributed over the myocardium. The density of these angiotensin II receptors is developmentally regulated, suggesting a possible role of the cardiac renin-angiotensin system in heart growth and in the adaptation of the heart to postnatal circulatory conditions. PMID- 8221743 TI - [Arterial hypertension and cognitive impairment]. PMID- 8221744 TI - Carvedilol. A wide therapeutic potential in cardiovascular syndromes. Proceedings of a satellite symposium organized by the International Institute for Therapeutic Research (ICIT). Scheveningen, The Netherlands, August 23, 1991. PMID- 8221745 TI - Infected health-care professionals: healers or modern day lepers? Part 1. Issues and considerations. PMID- 8221746 TI - Endodontic care: treatment vs emergency treatment vs palliative treatment. PMID- 8221747 TI - The implant-supported overdenture: a practical implant-prosthetic design. PMID- 8221748 TI - Designing occlusion for complete dentures. PMID- 8221749 TI - Cariology: theories past and present. PMID- 8221750 TI - Morbidity and mortality associated with the pharmacologic management of pain and anxiety. PMID- 8221751 TI - Use of endosseous implants to fixate a surgical stent in conjunction with mandibular vestibuloplasty. PMID- 8221752 TI - Dynamics of provisionalization in advanced periodontal prosthesis cases. PMID- 8221753 TI - A new material for periodontal splinting and orthodontic retention. PMID- 8221754 TI - Comparison of clinical and statistical models in age estimation using dental periapical radiographic parameters. PMID- 8221755 TI - Removable partial denture repairs. PMID- 8221756 TI - Endothelial cell induced modulation of cardiac fibroblast collagen metabolism. AB - OBJECTIVE: The aim was to examine the influence of vascular endothelial cells on rat cardiac fibroblast collagen synthesis and collagenase activity under co culture conditions, and to determine whether angiotensin II or aldosterone influence endothelial cell induced modulation of fibroblast collagen metabolism. METHODS: Bovine aortic endothelial cells were grown to confluence on microporous membrane inserts and then co-cultured with cardiac fibroblasts obtained from adult rats, previously grown to confluence in 35 mm dishes supplemented with 0.4% fetal bovine serum. Collagen synthesis was measured by 3H-proline incorporation. Experiments were also conducted under similar conditions to quantitate collagenase activity by zymography in conditioned medium from fibroblasts co cultured with endothelial cells. RESULTS: After 48 h co-culture, there was a 1.9 fold increase in fibroblast collagen synthesis when compared to fibroblasts alone (p < 0.001). Aldosterone (10(-8) M) or angiotensin II (10(-7) M) added to endothelial cells did not increase fibroblast collagen synthesis over co-cultures alone. Neither the aldosterone receptor antagonist spironolactone (10(-8) M) nor type I (DuP 753, 10(-8) M) or type II (PD 123319, 10(-8) M) angiotensin II receptor antagonists altered fibroblast collagen synthesis in co-cultures. A significant increment in collagenase activity was observed in co-cultured fibroblasts relative to collagenase activity of fibroblasts alone. CONCLUSIONS: Endothelial cells modulate both cardiac fibroblast collagen synthesis and degradation. The nature of the responsible signal(s) remains to be defined, but does not appear to be mediated or regulated by angiotensin II or aldosterone. PMID- 8221757 TI - Magnesium enhances function of postischaemic human myocardial tissue. AB - OBJECTIVES: The effect of Mg2+ on the developed force and concentrations of high energy phosphate metabolites in isolated human atrial trabeculae has been investigated. METHODS: Human atrial trabeculae, obtained from right atrial appendages of patients undergoing cardiac surgery requiring cardiopulmonary bypass, were dissected at room temperature in modified Krebs-Henseleit buffer containing 1.2 or 16 mM Mg2+, mounted on muscle stands, and rewarmed to 34 degrees C in the same buffer. After 30 minutes, their mechanical function was assessed. At the end of the protocol, trabeculae were fast frozen for measurement of concentrations of metabolites of high energy phosphates. RESULTS: Trabeculae collected and rewarmed in 16 mM Mg2+ Krebs-Henseleit buffer showed significantly higher mean developed force (0.59(SEM 0.10) g, p < 0.01) than those rewarmed in 1.2 mM Mg2+ Krebs-Henseleit buffer (0.32(0.03) g). Trabeculae that had a developed force > or = 0.8 g, a resting force < or = 0.7 g, and a cross sectional area < or = 1 mm2 ("functional" trabeculae) were selected for further comparison. New reverse phase high performance liquid chromatography techniques developed for the analysis of small samples (0.5-5 mg dry weight) were used to measure nucleotide, nucleoside, and creatine compounds. Total adenylate (ATP+ADP+AMP) concentrations in trabeculae revived in the presence of 16 mM Mg2+ (15.4(1.1) mumol.g-1 dry weight) were significantly higher (p < 0.01) than in those revived with 1.2 mM Mg2+ (11.8(1.0) mumol.g-1), but lower (p < 0.01) than in trabeculae fast frozen immediately after removal from the patient (22.6(1.0) mumol.g-1). There were no significant differences in NAD and total creatine (phosphocreatine+creatine) concentrations between the three groups. CONCLUSIONS: The presence of high Mg2+ during the rewarming of human atrial trabecular preparations maintains a significantly higher developed force and a significantly higher total adenylate pool than does collection and rewarming with normal concentrations of Mg2+. PMID- 8221758 TI - Inhibition of neutral endopeptidase 3.4.24.11 in conscious dogs with pacing induced heart failure. AB - OBJECTIVE: The effects of a selective neutral endopeptidase inhibitor, SQ 28,603 (N-[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]-beta-alanine), were determined in an experimental model of heart failure. METHODS: The symptoms of heart failure were induced by rapid ventricular pacing for one or three weeks in dogs with surgically implanted catheters for measurement of atrial pressures and mean arterial pressure and with ultrasonic flow probes for determination of cardiac output and renal blood flow. RESULTS: Inhibition of neutral endopeptidase by 10, 30, or 100 mumol.kg-1 SQ 28,603 given intravenously increased sodium excretion, cyclic GMP excretion, and plasma concentrations of atrial natriuretic peptide in a dose related manner in conscious dogs paced for one week. SQ 28,603 (100 mumol.kg-1) stimulated similar natriuretic and cyclic GMP responses in dogs paced for three weeks although baseline glomerular filtration rate was reduced. Because the natriuresis was maintained despite the smaller filtered sodium load, the increase in fractional sodium excretion was significantly greater after three weeks of pacing (from 0.5(0.2) to 3.7(0.7)%) than after one week of tachycardia (from 0.1(0.0) to 2.0(0.3)%). By contrast, SQ 28,603 (100 mumol.kg-1) did not affect renal, haemodynamic, or hormonal variables in normal conscious dogs where baseline atrial natriuretic peptide (18(3) fmol.ml-1) was lower than in the paced animals (104(10) fmol.ml-1). CONCLUSIONS: Inhibition of neutral endopeptidase in dogs with pacing induced heart failure protected endogenous atrial natriuretic peptide from degradation and stimulated sustained natriuresis, presumably via a tubular mechanism. PMID- 8221759 TI - Left ventricular energetics: heat production by the human heart. AB - OBJECTIVE: The aim was to examine the effect of coronary artery disease on human left ventricular energetics by a comparison of left ventricular oxygen consumption and heat production. The usefulness of measurement of left ventricular heat production for the detection of the expected change in left ventricular energetics produced by atrial pacing to a faster heart rate was also assessed. METHODS: Forty six patients (mean age 57 years; 31 men) undergoing cardiac catheterisation and coronary arteriography for the investigation of chest pain were studied. Normal left ventricular function and normal coronary arteries were present in eight and 38 had atheromatous coronary artery disease. Left ventricular heat production was calculated from coronary blood flow, the coronary arteriovenous (aorta-coronary sinus) temperature difference, and the areas under thermodilution curves recorded in the aorta and coronary sinus after injection of cold saline into the pulmonary artery. Mean external left ventricular power was calculated from mean arterial blood pressure and cardiac output. Left ventricular mechanical efficiency was derived from heat production and the energy value of myocardial oxygen use, assuming aerobic metabolism. In 27 patients studies were repeated during atrial pacing from the coronary sinus. RESULTS: At rest under basal conditions left ventricular heat production was 2.4(SD 1.0) W in patients with normal hearts and 3.1(1.4) W in patients with coronary disease (NS). Mechanical efficiency was 44.2(9.7)% in the normal patients and 30.7(10.9)% in those with coronary disease (p = 0.003). During atrial pacing to a faster heart rate left ventricular energy supply increased from 4.6(2.7) W to 5.9(3.3) W (p < 0.0005), and heat production increased from 3.0(1.6) W to 4.6(2.4) W (p < 0.0005), but mean external power was not altered. As the extra energy used during pacing was "wasted" as heat, there was a significant fall in left ventricular mechanical efficiency with pacing from 33.9(13.5)% to 18.9(15.2)% (p < 0.0005). CONCLUSIONS: These results show the effect of coronary artery disease on the energetics of left ventricular function. They also show that the method and equipment can detect the expected alteration in left ventricular energetics produced by atrial pacing. The measurement of left ventricular heat production and oxygen consumption allows assessment of the total left ventricular energy flux, and may be useful for the evaluation of drug treatment with such as inotropes and vasodilators, and for the investigation of the functional consequences of left ventricular disease. PMID- 8221760 TI - Acute pericardial distension in pigs: effect of fluid conductance on body surface electrocardiogram QRS size. AB - OBJECTIVES: Pericardial effusion has long been associated with low voltage of the surface electrocardiogram and its clinical utility is well known. The cause of this reduced QRS voltage has usually been attributed to short circuiting of the impulses by the pericardial fluid, but its precise mechanism has not been clearly elucidated. Therefore, we investigated this phenomenon by instilling various fluids with different resistances in increasing volumes into the pericardial space to finally produce acute cardiac tamponade. METHODS: Acute cardiac tamponade was produced 65 times in 25 closed chest pigs with a mean weight of 57(SEM 1.2) kg. A sufficient quantity (mean = 200 ml) of plasma, saline, blood, and blood with varying packed cell volume was introduced into the pericardial space in 60 ml increments to decrease systemic and pulmonary pressures to 50% of control to produce cardiac tamponade. At that point intrapericardial pressure had increased to 15-20 mm Hg. Resistivity values for the instilled fluids are 150 omega-cm for blood, 60 omega-cm for plasma, and 50 omega-cm for saline. RESULTS: Mean limb and precordial lead QRS voltage fell significantly, with no significant differences among these fluids. Electrocardiograph recordings from a unipolar electrode catheter in the right ventricle showed an increase in R wave voltage, but body surface recordings of stimuli introduced into the right atrium via a bipolar electrode catheter showed no amplitude change. CONCLUSIONS: In considering possibilities such as internal shunting of cardiac currents by intrapericardial fluid, distance of the heart from body surface electrodes, and change in cardiac position, the results are best explained by a reduction of cardiac volume and size during cardiac tamponade. This conclusion confirms Brody's hypothesis. PMID- 8221761 TI - Preventive effect of chronic converting enzyme inhibition on aortic stiffening induced by renovascular hypertension in conscious dogs. AB - OBJECTIVE: The aim was to assess the influence of the renin-angiotensin system on the geometrical and elastic properties of the aorta in conscious dogs, using a model of renovascular hypertension, and to examine the effects of inhibition of the system by the angiotensin converting enzyme inhibitor spirapril. METHODS: The aortic elastic behaviour in response to renovascular hypertension was studied in 15 conscious dogs instrumented with a pressure microtransducer and a pair of ultrasonic diameter dimension gauges in the upper descending thoracic aorta. Renovascular hypertension was induced by surgical occlusion of one renal artery and stenosis of the other. One day after renal surgery, dogs were randomly assigned to two groups receiving for two months either the new angiotensin converting enzyme inhibitor spirapril (n = 8) or a placebo capsule (n = 7). The two groups of dogs were compared to a control group of normotensive dogs (n = 7). After two months of treatment the elastic properties of the aorta were studied by computation of the beat to beat pressure-diameter hysteresis loops obtained during transient increase of pressure induced by bolus doses of angiotensin. The aortic pressure-diameter (P-D) relationship, obtained over a wide range, was fitted by an exponential fit (P = alpha.e beta D), where beta is the stiffness index. A decomposition of the P-D curve according to a biphasic model of the parallel arrangement of elastin and collagen enabled two pressure-diameter elastic moduli to be obtained, one representing the resistance to stretch at low pressure levels (elastic fibres and smooth muscle), and the other representing the resistance to stretch at the highest pressures (collagen fibres). RESULTS: The pressure-diameter curve of the placebo group was shifted to the left compared to the curves of the control and spirapril groups, showing that renovascular hypertension was associated with isobaric reduction of aortic diameter. The stiffness index beta was higher (p < 0.05) in the placebo group [0.605(SD 0.304) mm-1] than in either the control group [0.362(0.126) mm-1] or the spirapril group [0.348(0.083) mm-1], suggesting that renovascular hypertension was associated with aortic stiffening. The biphasic analysis showed that the collagen pressure diameter elastic modulus was unaffected by spirapril, whereas the elastin pressure-diameter elastic modulus was significantly reduced by converting enzyme inhibitor with respect to the placebo (p < 0.05). CONCLUSIONS: Chronic converting enzyme inhibition by spirapril prevents the isobaric aortic diameter reduction induced by renovascular hypertension in conscious dogs and decreases aortic stiffness, in particular by changing the elastic behaviour of the elastin fibres rather than of the collagen fibres. PMID- 8221762 TI - Effect of endogenous digoxin-like factor and digoxin antibody on myocardial Na+, K(+)-pump activity and ventricular arrhythmias in acute myocardial ischaemia in rats. AB - OBJECTIVE: The aim was to study whether a circulating sodium pump inhibitor (endogenous digoxin-like factor) contributes to the genesis of early ventricular arrhythmias in acute myocardial ischaemia in rats. METHODS: Effects of digoxin antibody (260 micrograms.kg-1) on the incidence of ventricular arrhythmias, plasma digoxin-like immunoreactivity (DELFIA immunoassay), Na+, K+, and Mg2+ ions, and activity of the ouabain sensitive Na+, K(+)-pump in different regions of myocardium have been studied in propranolol naive and propranolol pretreated rats exposed to acute coronary artery ligation. Adult male Wistar rats were divided into six experimental groups: (1) saline pretreated controls; (2) saline pretreated coronary artery ligated rats; (3) coronary artery ligated rats pretreated with 260 micrograms.kg-1 digoxin antibody; (4) propranolol pretreated controls; (5) propranolol pretreated rats with acute myocardial ischaemia; (6) rats with acute myocardial ischaemia pretreated with both propranolol and digoxin antibody. RESULTS: Acute myocardial ischaemia in saline pretreated rats was associated with a twofold increase of plasma digoxin-like immunoreactivity and ventricular arrhythmias, but did not lead to changes in myocardial Na+, K(+)-pump activity. Pretreatment of coronary artery ligated rats with digoxin antibody reduced the total duration of ventricular tachycardia and ventricular fibrillation during a 15 minute postligation period from 201 (SEM 34) to 46(18) seconds (p < 0.002) but did not alter activity of the myocardial Na+, K(+)-pump. In rats pretreated with propranolol, acute myocardial ischaemia was associated with a twofold inhibition of the Na+, K(+)-pump in left atrial and left ventricular myocardium, and with a 69% increase in plasma K+ concentration. Administration of digoxin antibody to propranolol pretreated coronary artery ligated rats in parallel with the antiarrhythmic effect prevented the increase in plasma K+ concentration and inhibition of Na+, K(+)-pump in the left atrial, but not the left ventricular myocardium. CONCLUSIONS: A circulating digoxin-like factor contributes to the pathogenesis of myocardial ischaemia induced ventricular arrhythmias. As propranolol pretreatment of coronary artery ligated rats inhibited the Na, K(+)-pump in myocardium, the inhibitory effect of endogenous digoxin-like factor on Na+, K(+)-ATPase was probably masked in propranolol naive animals by the stimulatory action of catecholamines on Na+, K(+)-ATPase described previously. PMID- 8221763 TI - Suppression of reperfusion induced arrhythmias in the isolated rat heart: pretreatment with 7-oxo prostacyclin in vivo. AB - OBJECTIVE: The aim was to investigate the late effect of pretreatment with 7-oxo prostacyclin on reperfusion induced arrhythmias in the isolated rat heart. METHODS: Forty eight hours after intramuscular administration of drug in vivo (50 micrograms.kg-1 body weight), isolated Langendorff perfused rat hearts were subjected to 30 minutes of regional ischaemia and 5 minutes of reperfusion. Incidence and duration of ventricular arrhythmias in both pretreated and control groups were evaluated on reperfusion. Morphological examination was also performed. RESULTS: In the untreated group reperfusion induced 75% of sustained ventricular fibrillation. Incidence of ventricular fibrillation, its duration, and arrhythmia score were significantly lower in the pretreated group. Pretreatment with 7-oxo prostacyclin had no effect on heart rate and coronary flow throughout the whole course of perfusion. Neither was the occluded zone size affected. Ultrastructure of ischaemic and reperfused myocardium was better preserved in the pretreated group. CONCLUSIONS: Antiarrhythmic action of 7-oxo prostacyclin was unrelated to changes in haemodynamics, thus suggesting the direct influence of the myocardium. The possible mechanism of action may involve maintenance of intracellular cation homeostasis (particularly of Na+ and Ca2+) due to a stimulation of sarcolemmal Na+ pump activity. PMID- 8221764 TI - Free radical generation coupled with arachidonate lipoxygenase reaction relates to reoxygenation induced myocardial cell injury. AB - OBJECTIVE: The role of arachidonate lipoxygenase activity in reoxygenation induced cell injury in adult canine cardiac myocytes was investigated. METHODS: The production of hydroxyeicosatetraenoic acids (HETEs), which are lipoxygenase metabolites, was measured with high pressure liquid chromatography in canine cardiac myocytes cultured under hypoxic conditions and then reoxygenated. Free radical generation was evaluated by electron paramagnetic resonance spectroscopy with a spin trapper, 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and luminol enhanced chemiluminescence emission. Cell injury was estimated in terms of morphological changes and release of intracellular enzymes. Morphological damage to myocytes was quantified in terms of the percentage of hypercontracted "round" cells. The effects of nordihydroguaiaretic acid, AA-861, mepacrine, indomethacin, aspirin, alpha tocopherol, and 2-0-octadecylascorbic acid (CV-3611) on lipoxygenase metabolism, free radical generation and cell injury were also assessed. RESULTS: Cardiac myocytes produced 5-HETE and 12-HETE at less than 0.1 ng.mg-1 protein under normoxic conditions. Production of HETE was greatly increased at five hours of reoxygenation after 45 minutes of hypoxia [5-HETE = 12.0(SEM 0.5), 12-HETE = 23.6(1.1) ng.mg-1 protein]. Both DMPO-OH adduct generation and chemiluminescence emission were considerably increased after one to three hours of reoxygenation, although they increased only slightly after 45 minutes of hypoxia. After five hours of reoxygenation, long rod cells gradually became deformed; 92.0% of the cells were converted to hypercontracted "round" cells. Cell injury and HETE production were significantly suppressed by nordihydroguaiaretic acid (10 microM), AA-861 (2 microM), and mepacrine (10 microM). Indomethacin (10 microM) and aspirin (50 microM) enhanced cell injury and HETE production. alpha Tocopherol and CV-3611 greatly suppressed cell injury and free radical generation, but not HETE production. CONCLUSION: The arachidonate lipoxygenase metabolic pathway may have an important role in reoxygenation induced myocardial cell injury in adult cardiac myocytes, possibly because of the generation of free radicals. PMID- 8221765 TI - Increased collagen synthesis and increased content of type VI collagen in myocardium of tight skin mice. AB - OBJECTIVE: The tight skin mouse (TSK) is a mutant strain characterised by excessive collagen accumulation in skin and some internal organs such as the heart. The aims of this study were (1) to examine in vitro collagen synthesis in TSK myocardium; (2) to determine myocardial content of type VI collagen in the same model; and (3) to examine the expression of one of the genes for type VI collagen in cultured TSK myocardial fibroblasts. METHODS: Individual hearts from 8 month old heterozygous male TSK and normal sex and age matched mice were incubated with 14C proline. Total 14C protein and 14C collagen synthesis by myocardial tissues were determined with collagenase digestion and SDS gel electrophoresis. For determination of type VI collagen, hearts from 10 month old male TSK and normal mice were subjected to guanidine extraction followed by pepsin digestion, salt fractionation, and western blotting. Expression of the alpha 2(VI) collagen gene was determined in myocardial fibroblasts cultured from TSK and normal mice, employing northern and dot blot hybridizations with a murine specific cDNA. RESULTS: TSK hearts had up to twofold greater protein and collagen biosynthesis and 2.5-fold greater type VI collagen content (400 micrograms v 156 micrograms). Fibroblasts cultured from TSK mice hearts displayed up to threefold higher steady state concentrations of alpha 2(VI) collagen mRNA than normal myocardial fibroblasts. CONCLUSIONS: Hearts from TSK mice showed increased protein and collagen biosynthesis and increased myocardial content of type VI collagen compared with hearts from age matched normal mice. Also, fibroblast cultures from TSK mice myocardium showed increased expression of the alpha 2(VI) collagen gene, indicating that increased transcription of type VI collagen genes may be responsible for the accumulation of this collagen in myocardium from TSK mice. PMID- 8221766 TI - Myocyte loss and left ventricular failure characterise the long term effects of coronary artery narrowing or renal hypertension in rats. AB - OBJECTIVE: The aim was to determine the effects of chronic coronary artery narrowing and two kidney, one clip renal hypertension alone and in combination on ventricular function and myocardial morphology. METHODS: Left coronary stenosis and renal artery clipping were surgically induced in rats and pump dynamics, systolic and diastolic wall stress, cardiac anatomy, and the changes in number and size of left ventricular myocytes were examined 11-13 weeks later. RESULTS: Ventricular failure evolved with each intervention: left ventricular end diastolic pressure was raised, whereas +dP/dt, -dP/dt, stroke volume, cardiac output, and cardiac index were reduced. Calculated ventricular systolic wall stress increased nearly 70% in the three experimental conditions. By contrast, diastolic wall stress was augmented 6.1-fold with coronary stenosis, 4.0-fold with hypertension, and 4.4-fold with combined treatment. These differences were due to variable preservations of wall thickness between the groups. Left ventricular weight expanded 26%, 35%, and 32% with stenosis, hypertension, and a combination of the two, whereas diastolic cavitary volume increased 57%, 35%, and 49%. Corresponding increases in systolic chamber volumes were 156%, 122%, and 154%. Finally, myocyte loss in the ventricle was 25%, 25%, and 37% in coronary narrowing, renal hypertension and a combination of the two with concomitant enlargements of the unaffected myocytes of 47%, 63%, and 65%. CONCLUSIONS: Decompensated eccentric ventricular hypertrophy developed as a result of coronary artery narrowing, renal hypertension, or the two in combination. Coronary artery narrowing, however, may have a greater maladaptive effect on ventricular function than systemic hypertension, and coronary stenosis and hypertension combined because of the more extensive chamber and wall remodelling which sustained greater increases in diastolic wall stress. PMID- 8221767 TI - Bronchial arteries in experimental pulmonary infarction: angiographic and morphometric study. AB - OBJECTIVES: The aim was to investigate (1) whether collateral bronchopulmonary circulation developing due to chronic pulmonary embolism could prevent the evolution of pulmonary infarction after induction of pulmonary venous outflow impairment; and (2) how collateral bronchopulmonary circulation developed after acute embolisation of the lung with impaired pulmonary venous outflow. METHODS: Fifty two mongrel dogs were studied. Thirty six dogs were experimental animals and 16 were in a control group. Unilateral impairment of pulmonary venous outflow was induced by constriction of the left pulmonary veins in two groups of experimental dogs: (1) three months after and (2) one hour before bilateral embolisation of the pulmonary artery. All animals were killed 12 days after constriction. The size of the bronchial arteries was evaluated from angiograms. The diameter and the wall thickness of the arteries were measured during histology. RESULTS: In all experimental dogs, haemorrhagic infarctions developed distally to emboli in the left lung regardless of whether the bronchial arteries were dilated before induction of pulmonary venous constriction or whether collateral circulation started to develop after pulmonary venous constriction. Constriction of the pulmonary veins was an essential factor for pulmonary infarction to develop as no infarction developed in the embolised regions of the right lungs with intact pulmonary venous outflow. Pulmonary venous constriction alone did not cause dilatation or hypertrophy of the bronchial arteries. After pulmonary artery embolisation, the same enlargement and hypertrophy of the bronchial arteries occurred both in the left lung with previously impaired venous outflow and in the right lung with intact pulmonary veins. CONCLUSIONS: Expanded bronchopulmonary circulation did not prevent the development of infarction in the embolised region of the lung with impaired pulmonary venous outflow. Development of collateral bronchopulmonary circulation was not influenced by previously impaired pulmonary venous outflow. PMID- 8221768 TI - Mechanical correlates of contraction-excitation feedback during acute ventricular dilatation. AB - OBJECTIVE: The purpose was to examine the mechanical correlates of the electrophysiological changes that occur during acute left ventricular dilatation. METHODS: Ten isolated, retrogradely perfused, ejecting rabbit hearts were studied. Left ventricular volume was adjusted by varying left atrial perfusion pressure. Left ventricular pressure was measured directly. Changes in left ventricular chamber dimensions at the level of an epicardial electrode were evaluated with two dimensional echocardiography and wall stress was calculated from these measures. Regional left ventricular electrophysiological properties were measured at two left atrial perfusing pressures. RESULTS: Increases in left atrial perfusion pressure resulted in significant increases in left ventricular end diastolic and end systolic pressures, epicardial and endocardial circumference, and wall stress. Only changes in diastolic wall stress correlated with changes in ventricular refractoriness (r = 0.69, p = 0.027). CONCLUSIONS: Left ventricular dilatation results in shortening of ventricular refractoriness in the isolated, ejecting rabbit heart. Regional changes in refractoriness are best correlated with changes in wall stress. PMID- 8221769 TI - The effects of exogenous lactate and pyruvate on the recovery of coronary flow in the rat heart after ischaemia. AB - OBJECTIVE: The effect of exogenous lactate and pyruvate on the recovery of coronary flow (total, regional) after ischaemia as a function of the duration of ischaemia was evaluated. METHODS: Isolated, ejecting rat hearts were subjected to ischaemia for 15, 30, or 45 minutes. Glucose (11 mM) was present as the basal substrate in the perfusion medium and lactate (5 mM) or pyruvate (5 mM) was added as the cosubstrate. Flow variables were measured by the timed collection of coronary effluents and by the radioactive microsphere technique. RESULTS: In the lactate perfused hearts, reactive hyperaemia was present after 15 minutes but absent after 30 and 45 minutes of ischaemia. Total coronary flow was significantly reduced after 45 minutes of ischaemia. Transmural flow was impaired after 15, 30, and 45 minutes of ischaemia in the lactate perfused hearts, -that is, flow in the inner layers of the left ventricle was transiently reduced after 15 minutes and remained continuously depressed after 30 and 45 minutes of ischaemia. The pyruvate perfused hearts showed reactive hyperaemia after 15 and 30 minutes of ischaemia. After 45 minutes total coronary flow was reduced below the value before ischaemia, and in particular, the inner layers of the left ventricle were severely deprived of flow as in lactate perfused hearts. When neither lactate nor pyruvate was added to the perfusion medium (containing glucose), impairment of coronary flow in the inner layers of the left ventricle was only transiently obvious after 30 and 45 minutes of ischaemia. Impaired perfusion of the inner layers during reperfusion resulted in delayed washout of lactate dehydrogenase. CONCLUSIONS: Exogenous substrates modify the recovery of flow after ischaemia. In the presence of exogenous lactate, severe disturbances of flow are already obvious after 30 minutes of ischaemia in the inner layers of the left ventricle. Exogenous pyruvate delays impairment of flow in the inner layers compared with exogenous lactate. PMID- 8221770 TI - Discrepancy between plasma and aortic wall cyclic guanosine monophosphate in an experimental model of congestive heart failure. AB - OBJECTIVES: The state of the vasodilator systems in congestive heart failure is poorly defined. Plasma atrial natriuretic peptide is increased, whereas endothelium derived relaxing factor activity can be decreased. Atrial natriuretic peptide and endothelium derived relaxing factor both cause vascular relaxation by generating cyclic guanosine monophosphate (cGMP), by activating the particulate and the soluble guanylate cyclase, respectively. This study examines the biological effects of atrial natriuretic peptide and endothelium derived relaxing factor in experimental heart failure by assessing the plasma, urinary, and tissue concentrations of their common second messenger cGMP. METHODS: Myocardial infarctions (n = 31) were induced and sham operations (n = 25) were performed on Wistar rats, and the rats were monitored for three months. Aortic and pulmonary cGMP contents were measured, as the aorta is mainly matrix and smooth muscle cells, and the lung is particularly rich in capillaries, hence in endothelial cells. The concentrations of the other second messenger cyclic adenosine monophosphate (cAMP) was also determined, as were those of cGMP dependent protein kinase in the arteries. RESULTS: 17 of the 31 rats with myocardial infarction had oedema. The total heart weight to body weight ratio and the ratio of the myocardium haemodynamically upstream from the infarcted left ventricle to body weight were increased in proportion to the infarct size. Plasma atrial natriuretic peptide and plasma and urinary cGMP concentrations were increased in proportion to the degree of heart failure (p < 0.0001). The pulmonary cGMP concentration was significantly higher in the rats with myocardial infarction than in the control group (p < 0.0001). Pulmonary cGMP concentrations were correlated with the plasma concentrations of atrial natriuretic peptide and cGMP (r2 = 0.59 and 0.66 respectively, p < 0.0001). The cGMP, cAMP, and cGMP, and cGMP dependent kinase concentrations in the aortic wall of rats with myocardial infarctions were the same as in control rats. CONCLUSIONS: The increase in plasma, urinary, and pulmonary cGMP in rats with myocardial infarctions were highly correlated with the increase in circulating atrial natriuretic peptide. By contrast, the aortic cGMP concentration was unchanged in these rats, despite high plasma atrial natriuretic peptide. In congestive heart failure, a discrepancy seems to exist between pulmonary (mainly endothelium) and aortic wall (mainly smooth muscle cells) cGMP. PMID- 8221771 TI - Cocultures of adult ventricular myocytes with stellate ganglia or intrinsic cardiac neurones from guinea pigs: spontaneous activity and pharmacological properties. AB - OBJECTIVE: The aim was to develop long term primary cocultures of adult ventricular myocytes with autonomic neurones dissociated from stellate or intrinsic cardiac ganglia. This was to provide an experimental model for the investigation of the regulatory role of autonomic neurones with respect to cardiac myocyte function. METHODS: Ventricular myocytes and stellate and intrinsic cardiac neurones were enzymatically dissociated from adult male guinea pigs and plated together on 13 mm cover slips; the cultures were maintained in an incubator for two to 10 weeks. The electrical properties of cultured myocytes and neurones were investigated by means of a conventional microelectrode technique and the spontaneous contractile activity of the myocytes was recorded by a video system. The electrical, contractile, and pharmacological properties of myocyte neuronal networks were investigated by superfusing the cultures with various neuromodulators and blockers. RESULTS: The electrical properties of the cultured myocytes and neurones were similar to those reported in other in vitro studies. Innervated and non-innervated cardiomyocyte cultures responded differently, however, to various pharmacological interventions. Spontaneous contractions were attenuated by tetrodotoxin (4 x 10(-7) M), beta adrenergic blockade, and nicotinic blockade more in cocultures than in cardiac myocyte cultures alone. On the other hand, the beta agonist isoproterenol increased the spontaneous beating in both types of cocultures more than in myocytes alone. The effects of the muscarinic agonist bethanechol and the muscarinic blocker atropine were not significantly different in innervated and non-innervated cultures. Nicotine induced either an increase or decrease in contractile rates of both cocultures and did not affect non-innervated myocyte cultures. CONCLUSIONS: Cocultures of myocytes and autonomic neurones alter the responses of myocytes relative to cultures without neurones. The results suggest that functional contacts can be established between adult myocytes and dissociated neurones in primary cultures. Contractile rates of such myocyte cultures are influenced by the presence of neurones. Neurones innervating these cultures were modified by beta adrenergic, muscarinic, and nicotinic agents. PMID- 8221772 TI - Skeletal muscle blood flow in heart failure measured by ultrafast computed tomography: validation by comparison with plethysmography. AB - OBJECTIVES: Abnormalities of skeletal muscle perfusion and metabolism may be important in the symptomatic limitation of patients with chronic heart failure. A method for assessing both skeletal muscle blood flow and mass would be useful in clinical practice and research. Ultrafast computed tomography has the potential to make these measurements. The aim was to determine the accuracy with which skeletal muscle blood flow could be measured by ultrafast computed tomography in patients with chronic heart failure. METHODS: Leg blood flow measured by venous occlusion plethysmography was compared with skeletal muscle blood flow by ultrafast computed tomography. Fourteen patients with chronic heart failure (aged 51 to 76 years) were investigated. Plethysmography and ultrafast computed tomography measurements were performed at rest and during hyperaemic flow induced by symptom limited bicycle exercise followed by five minutes of leg ischaemia. The ultrafast computed tomography measurements were made by analysing the opacification of the blood pool and of the muscle after an intravenous bolus of non-ionic radio-opaque contrast. RESULTS: Flows assessed by plethysmography ranged from 1.5 to 38.1 ml x 100 ml-1 x min-1. The slope of the line relating the two methods was 1.1 (95% confidence interval 0.91 to 1.31), and the mean (95% limits of agreement) of the differences between the two methods was 2.5(10.6) ml x 100 ml-1 x min-1. CONCLUSIONS: Ultrafast computed tomography is a useful tool in the measurement of both skeletal muscle mass and perfusion in humans. PMID- 8221773 TI - Mechanism of mitochondrial enzyme leakage during reoxygenation of the rat heart. AB - OBJECTIVE: The aim was to clarify the factors that induce enzyme release from mitochondria during anoxia and reoxygenation. METHODS: Isolated perfused hearts or isolated mitochondria were prepared from hearts excised from rats. The amounts of lactate dehydrogenase, cytoplasmic aspartate aminotransferase, and mitochondrial aspartate aminotransferase released into the coronary effluent from perfused heart preparations were measured. To distinguish the effect of mechanical stress from that of reoxygenation, a latex balloon was placed in the left ventricular cavity to impose mechanical stress and the heartbeat was controlled with a high K+ medium. A digitonin infusion technique was used to obtain only the cytosolic compartment of the cells for analysis of the amounts of mitochondrial enzymes released into the cytosol. The effect of anoxia followed by reoxygenation on enzyme release from isolated mitochondria was studied. RESULTS: On reoxygenation, mitochondrial aspartate aminotransferase was released as well as cytoplasmic enzymes, but, unlike cytoplasmic enzymes, the release was not influenced by mechanical stress. Mitochondrial injury by reoxygenation depended on the duration of the preceding anoxia. Reoxygenation of isolated mitochondria also induced enzyme release and the presence of ATP in the extramitochondrial space reduced the release of this enzyme. CONCLUSIONS: Enzyme leakage from mitochondria of myocardial cells occurs during reoxygenation, irrespective of mechanical stress, and this vulnerability to oxidative stress depends on the duration of the preceding anoxic period or the concentration of cytosolic ATP. PMID- 8221774 TI - Recovery of hypoxic neonatal hearts after cardioplegic arrest. AB - OBJECTIVES: Surgery for repair of congenital heart defects in the infant may be affected by hypoxia associated with the defect. The effects of chronic hypoxia on systemic ventricular function are not well characterised and few studies have considered myocardial preservation in the hypoxic neonatal heart. The aim was to determine how chronic hypoxia would affect left ventricular function in neonatal rabbit hearts subjected to global ischaemia. METHODS: Hearts from rabbits one, four, and six weeks of age and raised at 9% O2 were compared with hearts from rabbits raised in ambient air. Haemodynamic variables were measured with an isolated heart preparation before and after cardioplegic arrest. Creatine kinase was measured during reperfusion and myocardial oxygen consumption (MVO2) during ischaemia. RESULTS: At all ages, hypoxic hearts had significantly lower peak dP/dt and contractility index (dP/dt/left ventricular pressure (LVP)) than normoxic controls. After ischaemia and reperfusion, one week hypoxic hearts did not differ significantly in recovery from controls. Four week hypoxic hearts had significantly higher stroke volume and aortic flow, and six week hearts had significantly higher coronary flow than age matched controls. Contractility index did not show significant differences between hypoxic and control animals at any age. Hypoxic hearts released less creatine kinase in the coronary effluent during reperfusion than did control hearts of similar age. Six week hypoxic hearts had significantly higher MVO2 measured during the second administration of cardioplegia compared with six week control hearts but MVO2 did not differ significantly at one and four weeks of age. CONCLUSIONS: Despite reduced baseline function, chronically hypoxic immature rabbit hearts can recover from an ischaemic insult as well as age matched controls, with less evidence of myocardial necrosis. This parallels clinical findings in cyanotic infants. PMID- 8221775 TI - Continuous measurement of canine coronary blood volume change with alterations of heart rate. AB - OBJECTIVE: The aim was to examine the effects of heart rate on total coronary blood volume with pressure-type plethysmography in isolated and vasodilated canine hearts. METHODS: Nine hearts were excised from anaesthetised mongrel dogs (13.1-15.2 kg) and perfused with arterial blood of other dogs (17.0-29.0 kg). The venous blood returning to the right atrium and both ventricles was drained under constant negative pressure (-10 mm Hg). A thin latex balloon filled with water was inserted into the left ventricle to keep the intraventricular volume constant. The pressure difference between the cylinder into which the heart was placed and a compensation chamber was measured as a change in coronary blood volume while heart rate was altered from 120 beats.min-1 (control heart rate) to a target level (60, 90, 150, or 180 beats.min-1). RESULTS: The mean coronary blood volume change compared with that at control heart rate was 1.65(SEM 0.32) ml x 100 g-1 at 60 beats.min-1 (p < 0.005) and -0.74(0.20) ml x 100 g-1 at 180 beats.min-1 (p < 0.005) under the perfusion pressure of 70 mm Hg. The mean volume decreased with the increase in heart rate. Diastolic-systolic variations in coronary blood volume also decreased with an increase in heart rate, from 0.61(0.06) ml x 100 g-1 (60 beats.min-1) (p < 0.005) to 0.12 ml.100 g-1 (180 beats.min-1). Both mean change and variation were almost linear functions of R-R interval (r = 0.88 and r = 0.83). Lowering the perfusion pressure from 70 to 40 mm Hg diminished the changes in both mean and variation of the coronary blood volume. CONCLUSIONS: Tachycardia reduces the mean coronary blood volume and the diastolic-systolic variations in isolated vasodilated canine hearts. PMID- 8221776 TI - Recovery of hypoxic neonatal hearts after cardioplegic arrest. PMID- 8221777 TI - Taurine and the heart. PMID- 8221778 TI - Cardiovascular effects of neuropeptide Y: receptor interactions and cellular mechanisms. PMID- 8221779 TI - Role of Na+/H+ exchange in cardiac physiology and pathophysiology: mediation of myocardial reperfusion injury by the pH paradox. AB - Na+/H+ exchange, an electroneutral cotransport system, is activated by reperfusion of the ischaemic heart. While activation can restore intracellular pH following an acid load, the concomitant increase in intracellular Na+ can also aggravate existing derangements of ionic homeostasis, particularly with respect to calcium overload, and result in exacerbation and acceleration of tissue injury, a phenomenon which has been termed the pH paradox. In addition, Na+/H+ exchange has been shown to participate in the activation of both platelets and neutrophils, factors widely acknowledged to participate in ischaemic and reperfusion injury. All studies thus far reported (summarised in the table) have shown desirable and beneficial effects of Na+/H+ exchange inhibitors on various cellular processes which contribute to myocardial reperfusion injury. These multiple effects of Na+/H+ exchange inhibitors are unique and unmatched by any other group of pharmacological agents. They offer the hope of superior tissue protection and salvage, with limited potential for toxicity, following reperfusion protocols. We propose, therefore, that activation of the Na+/H+ exchanger mediates reperfusion injury and that suppression of the exchanger will be of superior benefit in reduction of such injury during restoration of flow. The rapid development of new and highly specific Na+/H+ exchange inhibitors offers substantial promise for the use of these agents as adjunct therapy in numerous reperfusion protocols. PMID- 8221780 TI - Oxygen radical scavenging agents as adjuvant therapy with tissue plasminogen activator in a canine model of coronary thrombolysis. AB - OBJECTIVES: Early thrombolysis can reduce infarct size and enhance the long term recovery of contractile function after acute myocardial infarction. These benefits of early reperfusion may be confounded, however, by platelet mediated reocclusion after initial lysis, and "reperfusion injury" mediated by oxygen derived free radicals. Superoxide dismutase (SOD)--as well as its action as a potent free radical scavenging agent--inhibits platelet aggregation in vitro. Thus our primary objectives were to determine whether SOD+catalase, given as adjuvant therapy with recombinant human tissue plasminogen activator, could inhibit platelet aggregation and thereby reduce the time to lysis and maintain arterial patency. Whether SOD+catalase enhanced myocardial salvage or improved acute recovery of contractile function in the setting of thrombosis/thrombolysis was also assessed. METHODS: Two anaesthetised open chest canine models were used: a model of thrombosis/thrombolysis and an in vivo model of platelet aggregation. In protocol I coronary thrombosis was induced by endothelial injury and injection of blood+thrombin+CaCl2. At three hours after occlusion, control animals received saline and tissue plasminogen activator and treated dogs received SOD+catalase and tissue plasminogen activator. Variables measured included coronary blood flow (for assessment of the time to lysis and incidence and duration of spontaneous reocclusion); regional myocardial blood flow; segment shortening; and infarct size. In protocol II dogs underwent endothelial injury and coronary stenosis, resulting in cyclic variations in coronary blood flow caused by formation and dislodgement of platelet thrombi. At 30 minutes after placement of the stenosis, dogs received either saline and tissue plasminogen activator or SOD+catalase and tissue plasminogen activator. Variables measured included the frequency of cyclic flow variations and the duration of occlusion per 30 minute time interval. RESULTS: SOD+catalase slightly reduced the time required to achieve reflow: time to lysis was 35(SEM 5) v 22(4) minutes for control v treated groups (p < 0.05). Protocol II showed that this accelerated time to lysis was due to acute inhibition of platelet aggregation by the scavenging agents. Despite continuous infusion however, SOD+catalase failed to maintain vessel patency in either limb of the study. In protocol I, reocclusion occurred in 90% of control dogs (with 49(8)% of the time after initial lysis spent reoccluded), v 83% in the treated group (36(12)% of the time reoccluded; NS v controls). Also, both groups remained dyskinetic after reflow: at one hour after initial lysis, segment shortening was 6(9)% v -6(20)% of baseline preocclusion values in the subset of control v treated dogs in which the left anterior descending coronary artery was patent at the time of measurement (NS). Finally, infarct size was similar in both groups, averaging 32(9)% v 38(9)% of the myocardium at risk in control v treated animals (NS). CONCLUSIONS: Although SOD+catalase acutely attenuated platelet aggregation and slightly accelerated lysis, these agents failed to limit platelet mediated rethrombosis. Furthermore, SOD+catalase did not enhance myocardial salvage and did not improve acute recovery of contractile function after thrombosis/thrombolysis. Thus SOD+catalase given as adjuvant therapy with tissue plasminogen activator did not have a substantial beneficial effect on either the efficacy of thrombolysis or on "reperfusion injury" in this canine model. PMID- 8221781 TI - Effects of heart rate on left ventricular filling dynamics: assessment from simultaneous recordings of pulsed Doppler transmitral flow velocity pattern and haemodynamic variables. AB - OBJECTIVE: The effects of heart rate on transmitral flow velocity pattern are still controversial. The aim of this study was to examine the relations between the changes in Doppler echocardiographic variables and those in haemodynamic variables in association with altering heart rate. METHODS: Pulsed Doppler transmitral flow velocity pattern and left atrial and left ventricular pressures were simultaneously recorded at various heart rates (from 80 to 120 beats.min-1) in 14 dogs. RESULTS: As heart rate increased, peak late diastolic filling velocity increased, the time-velocity integral of the early diastolic filling wave decreased, and the deceleration time of early diastolic flow shortened without significant changes in mean values of peak early diastolic filling velocity and acceleration time of early diastolic flow. Peak early diastolic filling velocity increased with heart rate, however, in five dogs and decreased in the other nine dogs. The shortening of the time constant of left ventricular isovolumic relaxation was greater in the five dogs in which peak early diastolic filling velocity increased with heart rate than in the other nine dogs. Results of multiple stepwise linear regression analysis showed that the changes in peak early diastolic filling velocity correlated with the changes in the crossover pressure and peak+dP/dt in addition to the time constant, and that diastolic filling period was not selected as an affecting factor. Changes in the other Doppler echocardiographic variables did not correlate solely with changes in heart rate, or diastolic filling period, but correlated in a complex fashion with the combined changes in haemodynamic variables associated with altering heart rate. CONCLUSIONS: Although transmitral flow velocity pattern changes with altering heart rate, this change is not considered to be a direct effect of alteration in heart rate, but an indirect one through the associated haemodynamic changes including left atrial pressure and left ventricular relaxation and contractility. PMID- 8221782 TI - Diminished arterial elasticity in diabetes: association with fluorescent advanced glycosylation end products in collagen. AB - OBJECTIVE: Non-enzymatic glycosylation of proteins occurs in diabetes and advanced glycosylated end products can accumulate in long lived proteins such as vascular collagen and reduce the elasticity of vessel walls. To evaluate the potential association of advanced glycosylated end products in collagen with diminished arterial elasticity in diabetes, 14 diabetic and 14 age and sex matched non-diabetic patients with coronary artery disease were studied. METHODS: Arterial elasticity was assessed in terms of carotid to femoral pulse wave velocity and by measuring the change in ascending aortic diameter induced by pulse pressure. Collagen linked fluorescence, a measure of advanced glycosylated end products, was determined from tissue specimens of the skin, ascending aorta, and right atrial appendage taken during coronary bypass surgery. RESULTS: As a sign of diminished arterial elasticity, carotid to femoral pulse wave velocity was raised (p < 0.01) and change in ascending aortic diameter tended to be diminished (p = 0.09) in the diabetic patients. Collagen linked fluorescence was increased (p < 0.05) in the myocardium of the diabetic group, but the difference in skin and aorta was not significant. Collagen linked fluorescence between the aorta, skin, and myocardium correlated with each other (r = 0.64-0.77). Collagen linked fluorescence in the aorta and myocardium correlated with carotid to femoral pulse wave velocity (r = 0.63 and r = 0.67, respectively) in the diabetic group but not in the control group. CONCLUSIONS: These data suggest that non enzymatic glycosylation of matrix proteins, and specifically collagen, may modify arterial elasticity in diabetic patients with coronary artery disease. PMID- 8221783 TI - Effects of temperature on cycle length dependent changes and restitution of action potential duration in guinea pig ventricular muscle. AB - OBJECTIVE: The aim was to investigate the effects of temperature on cycle length dependent changes of action potential duration and on restitution of action potential duration. METHODS: Guinea pig papillary muscle action potentials were recorded using conventional microelectrode techniques. Action potential duration was measured at cycle lengths ranging from 500 to 2000 ms at both 27 degrees C and 37 degrees C. Restitution of action potential duration was determined by introducing an extra stimulus at progressively longer diastolic intervals from 40 to 9000 ms at pacing cycle lengths of 500, 1000, and 2000 ms. RESULTS: At 37 degrees C, action potential duration measured at 90% of repolarisation (APD90) during continuous pacing and the maximum value of APD90 achieved during restitution (APD90pl) decreased by 18(SEM 6) ms (n = 7) and 24(7) ms (n = 6), respectively, when pacing cycle length was reduced from 2000 to 500 ms. At 27 degrees C, the magnitude of the shortening of APD90 and APD90pl observed when pacing cycle length was similarly reduced was greater than at 37 degrees C, ie, 143(21) ms (n = 6) and 115(11) ms (n = 6), respectively. Thus the relation for restitution of action potential duration shifted downwards with reduction in pacing cycle length, and the magnitude of this shift was greater at 27 degrees C than at 37 degrees C. The difference between APD90 at the shortest diastolic interval (40 ms) and at diastolic interval of 100 ms (range of premature action potential durations) was much greater at 27 degrees C than at 37 degrees C at all three pacing cycle lengths. CONCLUSIONS: Reduction in temperature magnifies the cycle length dependent changes in action potential duration both during abrupt changes in cycle length, as with an extra stimulus, and during changes of steady state cycle length. This may indicate a greater dispersion of premature action potential durations during hypothermia, and hence predispose to hypothermia induced arrhythmias. PMID- 8221784 TI - Medial necrosis and acute alterations in aortic distensibility following removal of the vasa vasorum of canine ascending aorta. AB - OBJECTIVE: It is known that the outer layers of the thoracic aorta receive substantial blood flow through vasa vasorum. This study was undertaken to test the hypothesis that removal of vasa vasorum flow will alter the elastic properties of the ascending aorta. METHODS: Distensibility of the ascending aorta was determined before and 30 min after careful removal of the periaortic fat network which contains the vasa vasorum in 10 acutely instrumented dogs (experimental group) and the results were compared with those obtained from six weight matched sham operated control dogs. Aortic distensibility was measured using the formula: distensibility = 2 x pulsatile changes in aortic diameter divided by (diastolic aortic diameter x pulse pressure). Aortic pressures were measured directly from the ascending aorta by a pressure gauge. Aortic diameters were simultaneously determined by an elastic air filled ring connected to a transducer. The efficacy of the technique for the interruption of vasa vasorum blood supply to the aortic wall was proved in six additional animals by histology of transverse blocks of aortic wall from the area of interest. Histology was performed before vasa vasorum removal in two animals, 30 min after vasa vasorum removal in another two, and 15 d after vasa vasorum removal in the remainder. RESULTS: At baseline, there was no difference in the measured variables between the two groups. Aortic distensibility decreased significantly in the experimental group after vasa vasorum removal by 0.90(SEM 0.17) 10(-6).cm2.dyn-1 (p < 0.001), while it remained unchanged in the control group during the experiment. Complete removal of vasa vasorum of the ascending aorta was found in experimental group animals which were killed 30 min after operation, while ischaemic medial necrosis was observed in those killed 15 d after operation. CONCLUSIONS: Vasa vasorum removal led to an acute decrease in the distensibility of the ascending aorta. Lack of blood supply to the outer part of the aortic wall is most likely to have accounted for these findings. PMID- 8221785 TI - Role of oxygen derived free radicals in the pathogenesis of coxsackievirus B3 myocarditis in mice. AB - OBJECTIVE: The aim was to test the role of oxygen derived free radicals in the development of myocarditis. This involved investigating the effects of polyethylene glycol conjugated superoxide dismutase (PEG-SOD, an enzyme catalysing the conversion of O2.- to H2O2) and polyethylene glycol conjugated catalase (PEG-catalase, accelerating the reaction of H2O2 to H2O and O2) upon coxsackievirus B3 (CB3) myocarditis. METHODS: Two week old male C3H/He mice were inoculated intraperitoneally with 10(3) plaque forming units of CB3. PEG-SOD, 1 x 10(3) U.kg-1 x d-1, and PEG-SOD, 1 x 10(3) U.kg-1 x d-1, plus PEG-catalase, 1 x 10(3) U.kg-1 x d-1, were injected subcutaneously daily on days 0 to 14. Treated groups were compared to the infected control. RESULTS: On day 7, there were no significant differences in pathological scores among the three groups. On day 14, the cellular infiltration, myocardial necrosis, and calcification scores were significantly lower in the PEG-SOD group and the PEG-SOD plus PEG-catalase group than in the control. There were no significant differences in pathological scores between the PEG-SOD group and the PEG-SOD plus PEG-catalase group. There were no differences in the myocardial virus titres on day 7 among the three groups. On day 14, virus was not detected from the myocardium in any of the three groups. CONCLUSIONS: The results suggest that superoxide anion is mostly responsible for myocyte injury in CB3 myocarditis in mice, and that hydrogen peroxide formed as a result of dismutation of superoxide anion may not play a significant role in the development of myocarditis. Superoxide anion is one of the most important factors in free radical mediated injury in CB3 myocarditis in mice and the administration of PEG-SOD alone has therapeutic potential in clinical CB3 myocarditis. PMID- 8221786 TI - Heat stress limits infarct size in the isolated perfused rabbit heart. AB - OBJECTIVE: Heat stress, with the expression of heat stress proteins, has been shown to protect the rabbit heart in vitro against global ischaemia/reperfusion injury, though no benefit is apparent in an in vivo rabbit model of infarct size. The aim of this study was therefore to investigate this discrepancy and to discover whether heating itself has any effect which could negate the protection derived from myocardial stress protein synthesis. METHODS: (1) To ascertain whether heat stress could limit infarct size in the absence of blood, isolated buffer perfused hearts, with or without prior heat stress, were subjected to 45 min of regional ischaemia and 120 min reperfusion, and the resulting infarct size was expressed as a percentage of the risk area (I/R%). (2) The observations were repeated in an isolated blood perfused heart model in which a support rabbit (heat stressed or control) was used to perfuse the isolated heart. RESULTS: In the buffer perfused heart, prior heat stress reduced I/R from 70.8(SEM 4.4)%, n = 10, in controls to 51.5(5.7)%, n = 12 (p < 0.05). In hearts perfused by support rabbits, prior heat stress reduced I/R [from 34.7(3.7)%, n = 16, to 23.5(3.3)%, n = 15 (p < 0.05)] only when the perfusing rabbit was a control (not heat stressed). If the perfusing rabbit had been heated, I/R was greater in both heat stressed and control hearts [51.9(7.0)% and 44.9(3.3)%, p < 0.05 v control support rabbit]. CONCLUSIONS: Heat stress limits infarct size in this rabbit model. However it appears to have additional adverse effects, probably on the blood, which may override any benefit associated with myocardial stress protein synthesis. PMID- 8221787 TI - Age related changes in contractility and alpha 1 adrenergic responsiveness of myocardium from normal hamsters and hamsters with hereditary cardiomyopathy. AB - OBJECTIVE: The aim was to evaluate age related changes in myocardial contractility and alpha 1 adrenergic responsiveness in myocardium from normal and cardiomyopathic hamsters. METHODS: 50 male cardiomyopathic hamsters (CHF 147) and 45 golden Syrian hamsters were killed at 30, 50, 100, 200, or 300 days of age. Left ventricular papillary muscles were isolated and mounted in an isolated bath and isometric, isotonic, and unloaded contractions were recorded during a phenylephrine dose-response curve. The effects of increasing stimulation rate from 6 to 45 x min-1 and of increasing temperature from 29 degrees C to 35 degrees C before and after the addition of phenylephrine were verified in 100 day old hamsters. RESULTS: At 30 days of age, tension generation and shortening indices were similar in both strains. Thereafter, contractility increased in normal hamsters, to reach a peak at 50 d which was maintained until 300 d, when a slight decrease occurred. In cardiomyopathic hamsters, contractility increased more slowly, such that by 50 d tension was lower than in normals, at 15.8(SEM 1.9) v 26.5(2.2) mN.mm-2 (p < 0.01). This decrease persisted until 200 d, when it normalised and then decreased again by 300 d. The response of normal and cardiomyopathic muscles to phenylephrine was similar at 30 days of age. Thereafter, the response of cardiomyopathic muscles decreased more than in normals. This decrease was accompanied by a rightward shift in the dose-response curve. Increasing stimulation rate had no effect under basal conditions; however, once phenylephrine was added it caused tension to increase in cardiomyopathic muscles instead of decreasing, thereby rendering the relative effects of phenylephrine at 45 stimuli.min-1 similar in both groups. CONCLUSIONS: Force generating and shortening indices in normal and cardiomyopathic hamster myocardium are similarly immature at 30 days of age. Normal myocardium reaches its full potential by 50 d, but cardiomyopathic myocardium does not reach it until 200 d, and it decreases thereafter. A decrease in alpha 1 adrenergic responsiveness in cardiomyopathic myocardium at lower stimulation rates can be reversed by increasing the stimulation rate. PMID- 8221788 TI - A stable model of left ventricular dysfunction in an intact animal assessed with high fidelity pressure and cinemagnetic resonance imaging. AB - OBJECTIVE: Numerous models of acute and chronic left ventricular dysfunction have been used over the years. However, few can produce a rapid onset of global systolic and diastolic dysfunction that is stable and potentially reversible. The aim of this study was to develop such a model. METHODS: A model of left ventricular dysfunction was produced in six intact dogs using 1% halothane anaesthesia and pharmacological autonomic blockade with atropine (0.1 mg.kg-1) and propranolol (2 mg.kg-1). Left ventricular function was assessed by combined high fidelity pressure and cinemagnetic resonance imaging (cine-MR) during increases in afterload using infusions of angiotensin. RESULTS: Left ventricular systolic dysfunction was characterised by a diminished resting ejection fraction of 45(SD 4)% and a depressed +dP/dtmax of 1537(100) mm Hg.s-1. Diastolic dysfunction was manifested by an increased left ventricular end diastolic pressure of 16(2) mm Hg, a decreased -dP/dtmax of -1705(369) mm Hg.s-1, and a prolonged time constant of left ventricular relaxation of 42(9) ms. As left ventricular systolic pressure steadily rose with angiotensin infusion from 87(7) to 124(13) to 152(10) mm Hg (p < 0.001), left ventricular ejection fraction decreased markedly from 45(4) to 35(4) to 27(4)% (p < 0.001). Left ventricular +dP/dtmax did not change [1537(100) to 1500(110) to 1498(84) mm Hg.s-1] in spite of a significant increase in left ventricular end diastolic pressure from 16(2) to 21(5) to 29(7) mm Hg (p < 0.001) and left ventricular end diastolic volume from 59(12) to 71(14) to 78(17) ml (p < 0.001). Individual slopes of the end systolic pressure-volume relationship were also low, ranging between 2.1 and 4.4 mm Hg.s-1 (r = 0.99 to 1.00), typical of impaired contractility. CONCLUSIONS: Halothane anaesthesia in dogs pretreated with large amounts of propranolol and appropriate muscarinic cholinergic blockade produces a moderate decrease in baseline systolic and diastolic function in our intact dog model. However, left ventricular systolic function showed limited contractile reserve when challenged by physiological increases in systemic arterial pressure. Impaired systolic and diastolic function may, at least in part, be related to diminished activator calcium produced by halothane in addition to the well known negative inotropic action of beta adrenergic blockade. PMID- 8221789 TI - Muscarinic receptor stimulation by carbachol improves functional recovery in isolated, blood perfused rabbit heart. AB - OBJECTIVE: The aim was to test whether, besides adenosine A1 receptors, other receptors coupled to the "inhibitory" GTP binding protein Gi confer protection against ischaemic and reperfusion injury. METHODS: Isolated hearts were Langendorff perfused at 37 degrees C with blood and were divided into five groups, all subjected to 45 min zero flow normothermic ischaemia, followed by 1 h reperfusion. Group 1 consisted of control untreated hearts (n = 5). Group 2 consisted of hearts treated with the muscarinic agonist carbachol (CCh, 10 microM; n = 5). In group 3 (CCh+atropine, n = 5) 5 microM atropine was given as well as CCh. Groups 1, 2, and 3 were paced at 3 Hz except during ischaemia. Groups 4 (n = 5) and 5 (n = 6) were similar to groups 1 and 2, except that pacing was continued throughout ischaemia. RESULTS: In group 1, left ventricular end diastolic pressure increased during ischaemia, from 7.2(SEM 1.2) mm Hg to 57(5.6) mm Hg, and remained increased during reperfusion, whereas in group 2, it increased moderately during ischaemia, from 9.2(0.9) mm Hg to 21.6(4.9) mm Hg and returned to preischaemic levels during reperfusion. In hearts paced during ischaemia, contracture developed faster than in unpaced hearts, but CCh still delayed contracture. Postischaemic recovery of isovolumetric left ventricular pressure or its first derivative (dP/dt) and reactive hyperaemia were significantly greater in CCh treated hearts [left ventricular pressure = 69.8(4.9)% of preischaemic value after 60 min reperfusion] than in control hearts [29.3(6.8)%]. Recovery of left ventricular pressure and dP/dt was worsened by pacing, but CCh still improved this recovery [left ventricular pressure = 13.1(8.4)% in control, 46.7(5.3)% in CCh treated hearts]. The CCh effect could be prevented by atropine. ATP breakdown and lactate accumulation during the first minute of ischaemia were lower in non-paced CCh treated hearts than in controls. ATP recovered better at the end of reperfusion in both non-paced [7.1(0.72) mumol.g-1] and paced [4.4(0.77) mumol.g-1] CCh treated hearts, as compared with non-paced [1.8(0.58) mumol.g-1] and paced [1.2(0.39) mumol.g-1] control hearts. In paced hearts, less creatine kinase and lactate dehydrogenase were released in the CCh treated group throughout the reperfusion phase. CONCLUSIONS: Carbachol improves functional recovery and confers cellular protection, and this protection depends mainly but not entirely on its bradycardic effect. PMID- 8221790 TI - Effects of nisoldipine upon endothelial dysfunction following ischaemic and peroxidative injury in the perfused rat heart. AB - OBJECTIVE: Previous studies showed that treatment of hearts with nisoldipine improves recovery of cardiac function following ischaemia, improves reperfusion, and reduces the constrictor sensitivity to endothelin. The aim of the present study was to assess the reduction in vasodilator responses that occurs following ischaemia and reperfusion or after oxidative stress, and to determine whether nisoldipine treatment improves these dilator responses. METHODS: Isolated perfused rat hearts were studied. Coronary vessels were constricted by the addition of U46619 and dilator responses were determined with the addition of acetylcholine (endothelium dependent) or glyceryl trinitrate (endothelium independent). Responses were compared before and after low flow ischaemia (20%, 30 min) and reperfusion (30 min), or treatment with tert-butyl hydroperoxide (tBHP) (0.9 mM, 12 min, 30 min wash). RESULTS: The addition of U46619 caused a prolonged increase in perfusion pressure of 50 to 70 mm Hg which was not significantly different before and after treatment. Dilatation responses to acetylcholine were significantly reduced following ischaemia and reperfusion (34% of preischaemic values) or tBHP (47.6% of pre-tBHP values), while responses to glyceryl trinitrate were not significantly changed. In contrast, when hearts were perfused with nisoldipine, the responses to acetylcholine were significantly improved (88% of preischaemic values with 5 nM nisoldipine, and 68% to 78% of pre tBHP values with 0.5 nM to 5 nM nisoldipine). Responses to acetylcholine following tBHP were not significantly improved when hearts were perfused with verapamil (5 nM 43.5%, 5 microM 32%, of pre-tBHP values), or diltiazem (5 nM 37%, 5 microM 31%, of pre-tBHP values). CONCLUSIONS: Ischaemia and reperfusion or oxidative stress reduced endothelium dependent responses, but not endothelium independent responses. Nisoldipine reduced the injury to endothelial cell function associated with ischaemia and reperfusion or oxidative stress. PMID- 8221791 TI - Analysis of cardiac assistance by latissimus dorsi cardiomyoplasty with a time varying elastance model. AB - OBJECTIVE: The clinical use of skeletal muscle cardiomyoplasty is limited because of its inadequate haemodynamic benefits. To facilitate experimental and clinical efforts to improve the efficacy of this technique, a mathematical model was proposed and its validity was tested in acute experiments. METHODS: The model was based on the assumption that the skeletal muscle wrapped around the heart behaves as a time varying elastance that is connected in series with another time varying elastance representing the native heart. From this model two predictions were made: (1) Skeletal muscle augments the contractility of the heart by increasing the slope (Ees) of the end systolic pressure-volume relation; (2) time varying elastance of the skeletal muscle chamber (Es(t)) can be estimated from that of the assisted heart. These predictions were examined in experiments. In nine anaesthetised, open chest dogs, preconditioned latissimus dorsi muscle was transposed to wrap the heart. Left ventricular pressure (catheter tipped micromanometer), and volume (conductance catheter) were measured while reducing the preload by vena caval occlusion to evaluate Ees with 1:2 (stimulation:heart beat ratio) stimulation of the skeletal muscle. RESULTS: With the stimulation of latissimus muscle, the end systolic pressure-volume relation was linear and Ees increased from 8.6(SEM 2.4) to 11.9(SEM 3.4) mm Hg.ml-1. Estimated Es(t) reflected the stimulation pattern and could account for the mechanism of the cardiac assistance. CONCLUSIONS: Skeletal muscle cardiomyoplasty improved the haemodynamic variable (Ees) as predicted by a mathematical model. PMID- 8221792 TI - Ischaemia and the myocyte cytoskeleton: review and speculation. AB - The cardiomyocyte cytoskeleton is composed of a highly organised complex array of specific proteins, arranged to transmit mechanical forces within the cell, to adjacent cells and the extracellular matrix, as well as to maintain internal organisation of cellular organelles. Although most of the published reports on cytoskeletal proteins refer to non-myocyte and smooth muscle cells, there seem significant homologies with cardiac structures. The specific interactions of some proteins in certain cytoskeletal structures are established and may be analogous to interactions in cardiac myocytes, but the roles of many proteins are uncertain, and the list of proteins that compose the cytoskeleton is likely to be incomplete. Some proteins may serve a dual role, contributing to signal transduction as well as to organisation and mechanical stability of the cell. Phosphorylation of cytoskeletal proteins, and elaborate cellular systems to control protein phosphorylation levels, suggest phosphorylation as a potential mechanism of controlling cytoskeletal assembly and remodelling. Disturbances of the cytoskeleton during ischaemia may produce alterations in cell structural integrity that could account for cell injury and death. Although mechanisms both of cytoskeletal assembly in normal cells and of cytoskeletal injury in ischaemic cells are currently poorly understood, research into the interactions of cytoskeletal proteins during ischaemia includes new approaches that may increase our understanding of the pathophysiology of the cardiac myocyte. PMID- 8221793 TI - Skeletal muscle assistance in heart failure. PMID- 8221794 TI - Pulsus alternans. PMID- 8221795 TI - Role of platelet activating factor in ischaemia-reperfusion injury of isolated rabbit hearts: protective effect of a specific platelet activating factor antagonist, TCV-309. AB - OBJECTIVES: The aims were to confirm that platelet activating factor is released during reperfusion after global ischaemia in isolated blood perfused rabbit hearts, and to examine the protective action of TCV-309, a platelet activating factor antagonist, against reperfusion injury of cardiac muscle. METHODS: The hearts were mounted on a Langendorff apparatus and perfused with diluted blood perfusate. After cardiac arrest with St Thomas's cardioplegic solution, the hearts were subjected to global ischaemia for 120 minutes at 25 degrees C, and then reperfused for 60 minutes at 37 degrees C. Release of platelet activating factor into the coronary effluent was quantified by radioimmunoassay. The effect of TCV-309 on left ventricular function and release of creatine kinase was measured. RESULTS: A pronounced release of platelet activating factor occurred after the commencement of reperfusion, although it was not detectable before induction of ischaemia. Release of platelet activating factor occurred over 60 minutes of the reperfusion period. In the control, left ventricular developed pressure after 60 minutes of reperfusion recovered to 54.3(SEM 1.7)% (n = 5) of the preischaemic value. In the hearts treated with TCV-309 at concentrations above 0.3 microM, recovery of left ventricular developed pressure was significantly improved (77.6(2.0)% at 1 microM, p < 0.01 v control). Leakage of creatine kinase during the initial five minutes of reperfusion was significantly less in the hearts treated with 1 microM TCV-309 than in the controls (5.2(0.4) v 12.2(1.4) IU.g-1 wet weight, p < 0.01). CONCLUSIONS: Release of platelet activating factor occurred during the reperfusion period in Langendorff perfused hearts. Treatment with the platelet activating factor antagonist TCV-309 significantly improved postischaemic left ventricular function and decreased creatine kinase release. These results suggest that platelet activating factor is involved in myocardial injury during ischaemia-reperfusion. PMID- 8221796 TI - Effects of inotropic stimulation on phosphate compounds in ischaemic canine hearts. AB - OBJECTIVE: The aim was to evaluate regional coronary blood flow, contractile function, and concentration of phosphate compounds with inotropic stimulation in moderately ischaemic canine hearts. METHODS: Dogs were prepared with instrumentation for the determination of regional coronary blood flow (non radioactive microsphere method), contractile function (sonomicrometry), and haemodynamics. Myocardial phosphate compounds were measured simultaneously by the phosphorus-31 magnetic resonance spectroscopic technique. RESULTS: In the non ischaemic condition, dobutamine increased regional coronary blood flow and enhanced contractile function with no significant changes in myocardial phosphate compounds. After constricting the left anterior descending coronary artery, dogs were divided into two groups according to their heart beat response to dobutamine, classified as no pronounced tachycardia with dobutamine (group NT), and pronounced tachycardia (group T). In group NT, dobutamine increased regional coronary blood flow and improved regional contractile function with no significant effect on myocardial phosphate compounds. In group T, dobutamine failed to increase regional coronary blood flow or to improve contractile function, but there was a significant increase in the inorganic phosphate to creatinine phosphate ratio. CONCLUSIONS: Dobutamine increased coronary blood flow and augmented contractile function without significant changes in phosphate compounds in the moderately ischaemic heart, when pronounced tachycardia was not induced. The augmentation of contractile function occurred without prominent improvement in energy metabolism. PMID- 8221797 TI - Differential sensitivity of proximal and distal coronary arteries to a nitric oxide donor following reperfusion injury or inhibition of nitric oxide synthesis. AB - OBJECTIVE: The effect of the nitric oxide donor, SIN-1, in proximal and distal coronary arteries with normal endothelium was characterised before and after inhibition of NO synthesis with L-nitroarginine methyl ester (L-NAME). The effect of reperfusion injury in vivo in similar vessels on the response to SIN-1 was also assessed. METHODS: In vitro reactivity of preconstricted coronary arterial rings was studied in control dogs (group 1), and dogs in which the left circumflex coronary artery was subjected in vivo to four acute occlusions of 5 min duration, with three intervening reperfusion periods of 5 min and a final reperfusion period of 60 min (group 2). The effects of acetylcholine and SIN-1 on the tone of left circumflex and left anterior descending coronary vascular rings were examined before and after treatment with L-NAME. RESULTS: Proximal [1851 (SEM 82) microns] and distal [477(19) microns] vessels were studied. In control dogs (group 1) acetylcholine caused relaxation in proximal and distal coronary arteries (p > 0.05). No difference in responsiveness of left circumflex or left anterior descending coronary arteries was observed in the control group. In group 2 the response to acetylcholine was significantly (p < 0.05) attenuated in left circumflex coronary arteries exposed to ischaemia and reperfusion compared with left anterior descending control rings from the same heart. Proximal vessels in group 1 and group 2 showed greater sensitivity to the vasodilator effects of SIN 1 than distal vessels. Proximal left circumflex vessels exposed to ischaemia and reperfusion showed enhanced sensitivity to the relaxant effects of SIN-1 compared to control proximal vessels obtained from the same hearts. Reperfusion was not associated with any alteration in sensitivity of distal vessels to SIN-1. Similarly, inhibition of the synthesis of endothelium derived relaxing factor (EDRF) by L-NAME resulted in an enhanced response to SIN-1 in proximal vessels only. CONCLUSIONS: Endothelium dependent vasodilatation is attenuated by ischaemia and reperfusion in both proximal and distal coronary arteries of the size studied. The response to direct nitric oxide donation (bypassing vascular endothelial synthesis of EDRF) is inhibited by a basal endothelial process present in proximal coronary arteries only. This inhibition is abolished following reperfusion injury or inhibition of NO synthesis. PMID- 8221798 TI - The contractile mechanism as an approach to building left ventricular pump models. AB - OBJECTIVE: The aim was to construct a model linking a simplified interpretation of the contractile process at the myofilament level to the mechanical behaviour of the left ventricle to improve the ability of elastic-resistive models to represent the pumping response of the left ventricle. The mechanical model, consisting of an elastic component connected in series with a contractile component and an elastic component parallel to both the series elastic and contractile components, is able to develop pressure by the binding of a structural substance T to an excitatory substance C, the behaviour of which is a simplification of miofibrillar Ca2+ kinetics. METHODS: Theoretically, the model was validated for its ability to reproduce by computer simulation, experiments that described the pumping properties of the left ventricle--namely, elasticity, resistivity, deactivating and positive effect of ejection, and the behaviour of intracellular Ca2+. Experimentally, the model was tested to fit intraventricular pressure (P(t)) and volume (V(t)) of single ejective beats in nine open chest dogs fitted with a pressure microtransducer to measure intraventricular P(t) and an aortic flowprobe to measure ventricular outflow and calculate V(t). Parameters were estimated up to maximum negative dP/dt adjusting P(t) or V(t) data of the ejective beats, and the goodness of the fit was evaluated through the root mean square error normalised with respect to the corresponding mean P(t) or V(t) in the fitting interval (NE). RESULTS: Descriptive validation of the model showed that the mean NE for the ejective P(t) fit was 0.03(SD 0.005) and for the V(t) fit 0.014(0.003). Predictive validation of P(t) and V(t) data of beats with partial occlusion of the aorta was performed up to end ejection, with parameters estimated from the P(t) or V(t) fit of the preceding ejective beat. Results gave a mean NE equal to 0.05(0.02) for predicted P(t) and 0.02(0.007) for predicted V(t), from either source of estimated parameters. Explanative validation showed that all the estimated parameters were in the same range used in simulation and that derived indexes [isovolumic maximum pressure (Pmax) = 166(13) mm Hg, time to maximum pressure (TPmax) = 0.186(0.012) s and the slope of the end systolic pressure volume relation (Emax) = 5.45(1.5) mm Hg.ml-1] were within reported experimental values. Finally, the model responded to increased inotropic state [dobutamine (5-35 micrograms.kg-1.min-1)] causing the estimated Pmax and Emax to increase by 33% and 25%, respectively, and TPmax to decrease by 10%. CONCLUSION: This model represented an improvement over previous pump models because (1) the model was able to represent behaviours other than purely elastic-resistive ones, such as the deactivation and positive effect of ejection; (2) left ventricular properties were the response of model behaviour and not constitutive elements of its structure; and (3) it adequately fulfilled model validation procedures. PMID- 8221799 TI - Ventricular coupling via the pericardium: normal versus tamponade. AB - OBJECTIVE: The aim was to examine how regional variations in pericardial pressure affect the mechanical coupling between the ventricles. METHODS: Canine hearts from 14 dogs (14.5-18 kg) were removed and placed in cold cardioplegia solution. Balloons were inserted into the left and right ventricles and the atria. Pericardial pressure over the left ventricle (Pclv) and the right ventricle (Pcrv) was measured with thin balloon catheters. Ventricular and pericardial pressures were measured, and ventricular and pericardial coupling was calculated, under control conditions and with increases in pericardial tension and fluid. RESULTS: At baseline, regional differences in pericardial pressure occurred [Pclv > Pcrv, 4.0(SD 0.9) v 2.9(0.6) mm Hg, p < 0.05]. Ventricular coupling via the pericardium was defined as delta Pclv/delta Pcrv for right ventricular volume increases and delta Pcrv/delta Pclv for left ventricular volume increases. This ratio increased more after increasing right ventricular volume than after increasing left ventricular volume [delta Pclv/delta Pcrv > delta Pcrv/delta Pclv, 1.14(0.33) v 0.51(0.15), p < 0.05]. Increasing the pericardial tension by clamping the pericardium increased pericardial pressures, yet did not alter the regional variations in pressure [Pclv > Pcrv, 8.4(2.2) v 6.4(2.5) mm Hg, p < 0.05] or pericardial coupling [delta Pclv/delta Pcrv > delta Pclv/delta Pcrv, 1.18(0.46) v 0.54(0.16), p < 0.05]. In contrast, creating a mild tamponade increased pericardial pressures, eliminated regional differences in pressure, and altered the coupling between ventricles [delta Pclv/delta Pcrv approximately delta Pclv/delta Pcrv, 0.95(0.11) v 1.05(0.08), p = NS]. These regional differences in pericardial pressure might have a geometrical basis. In four in vivo canine experiments using cine magnetic resonance, the short axis radius of curvature for the right ventricle was greater than for the left ventricle [38.3(4.4) mm v 29.2(3.8) mm, p < 0.05]. CONCLUSIONS: The pericardium partially protects right ventricular filling: regional differences in pericardial pressure normally occurred with lower pericardial pressure over the right ventricle, and left to right ventricular coupling was less. This protection of right ventricular filling was lost with even a small pericardial effusion. PMID- 8221800 TI - Effect of intravenous calcitonin gene related peptide on ischaemia threshold and coronary stenosis severity in humans. AB - OBJECTIVE: Calcitonin gene related peptide (CGRP) is a potent vasodilator found in sensory nerve endings in coronary arteries. The effect of intravenous CGRP on myocardial ischaemia, cardiovascular haemodynamics, and epicardial coronary artery stenoses was studied in patients with angina to investigate the possible role of CGRP in the control of vasodilation in the coronary circulation. METHODS: 12 patients (mean age 60 years) performed exercise tests on consecutive days during intravenous infusions of placebo or CGRP (at 50 pmol.min-1). At cardiac catheterisation (6 patients), consecutive 20 min intravenous infusions of placebo and CGRP were given at a fixed heart rate, with measurement of haemodynamic variables and quantitative arteriography. RESULTS: The time to 0.1 mV ST depression increased from 475(SD 245) to 546(193) s (p < 0.05), and time to 0.2 mV ST segment depression, from 634(266) to 694(202) s (p < 0.05), on placebo and CGRP respectively. Rate-pressure product at 0.1 mV ST segment depression was 18,410(4590) and 20,750(7270) mm Hg.min-1 (p < 0.05), and at 0.2 mV ST depression it was 21,510(6470) and 24,080(7380) mm Hg.min-1 (p < 0.01), with placebo and CGRP, respectively. At catheterisation, diastolic blood pressure fell from 93(11) to 83(8) mm Hg (p < 0.05), systemic vascular resistance fell from 26.2(9.4) to 20.7(9.0) units (p < 0.05), and cardiac output increased from 5.94(1.10) to 6.73(1.04) litre.min-1 (p < 0.05), on placebo and CGRP respectively. Minimum luminal diameter increased from 0.66(0.23) to 1.11(0.15) mm (p < 0.01) with CGRP. CONCLUSIONS: Intravenous CGRP is a systemic arterial vasodilator which dilates coronary arteries at the site of atheromatous stenoses and delays the onset of myocardial ischaemia during treadmill exercise testing in patients with chronic stable angina. PMID- 8221801 TI - Release of endothelin from the porcine heart after short term coronary artery occlusion. AB - OBJECTIVE: Endothelin is increased in plasma following myocardial infarction. Whether brief periods of myocardial ischaemia not leading to myocardial infarction increase plasma endothelin is not known. Thus, the present study was designed to examine cardiac endothelin balance in association with a 10 min coronary artery occlusion followed by reperfusion. METHODS: Venous blood was selectively sampled from the transiently ischaemic myocardium using a shunt between the anterior interventricular vein and the right atrium in eight pentobarbitone anaesthetised pigs. Flow in the shunt was measured with a Doppler flow probe. Arterial blood was drawn from the aortic arch. Plasma endothelin was measured using an Endothelin 1-21 specific [125I] assay system. This assay system has no cross reactivity with big endothlin. RESULTS: A net cardiac endothelin uptake of 0.7(0.3-1.4) fmol.min-1 x g-1 (median, 95% confidence interval) in the control period shifted to a net release during the first 10 min of reperfusion. The release reached a maximum of 2.8(0.4-6.0) fmol.min-1 x g-1 after 1.5 min of reperfusion. Cardiac venous endothelin concentration increased from 3.4(2.5-4.8) to 4.4(3.6-6.9) and 4.4(3.6-6.6) fmol.ml-1 at 1.5 and 5 min of reperfusion, respectively (p < 0.001 for both). Arterial endothelin concentration decreased from 4.8(3.9-6.1) to 2.7(2.4-4.3) fmol.ml-1 at 10 min of reperfusion (p < 0.001). CONCLUSION: Endothelin is released from the heart for several minutes during reperfusion following a brief coronary artery occlusion. PMID- 8221802 TI - Interleukin-1 beta modulates myocardial contraction via dexamethasone sensitive production of nitric oxide. AB - OBJECTIVE: Nitric oxide released by a calcium dependent constitutive NO synthase in endocardial endothelial cells exerts characteristic effects on myocardial contraction. Interleukin 1 beta (IL-1) induces the expression of a different calcium independent NO synthase in several tissues. Activity of the latter enzyme has recently been identified in cardiac myocytes but its functional effects are unknown. The aim of this study was to investigate the effects of IL-1 on contraction of isolated ferret papillary muscle preparations. METHODS: Electrically stimulated preparations were studied in the presence of acebutolol (1 microM), indomethacin (10 microM), and polymyxin (10 micrograms.ml-1). After a 3 h equilibration period, IL-1 (10 ng.ml-1) was added and contractile behaviour monitored over the next 3 h. The following groups were studied: (1) no IL-1; (2) IL-1 alone; (3) IL-1 in the presence of dexamethasone (3 microM); (4) IL-1 in the presence of NG-monomethyl-L-arginine (L-NMMA, 50 microM); (5) IL-1 in the presence of L-NMMA (50 microM) and L-arginine (500 microM). Cyclic GMP content was measured by radioimmunoassay in all preparations. RESULTS: No significant contractile changes were noted in group 1. In group 2, IL-1 raised myocardial cyclic GMP and induced a progressive abbreviation of isometric twitch, starting 30 min after addition, with a small decrease in peak isometric tension but no change in rate of tension development. These effects were similar to those previously documented for endocardial endothelial NO but were not inhibited by endocardial endothelial denudation. IL-1 effects were inhibited in groups 3 and 4, and partially restored in group 5. CONCLUSIONS: IL-1 activates a dexamethasone sensitive myocardial L-arginine-NO pathway which raises myocardial cyclic GMP and induces marked twitch abbreviation. These effects could potentially lead to cardiac depression by limiting systolic ejection, and may be relevant in conditions where IL-1 levels are increased. PMID- 8221803 TI - R56865, an antiarrhythmic drug with class III effects that terminates ouabain induced ventricular tachycardia in an inverse rate dependent manner. AB - OBJECTIVE: R56865 has been described as a substance that protects cells from intracellular Na+ and Ca2+ overload. The aim of this study was to investigate its mechanism of action, which is at present unknown. METHODS: The haemodynamic and (rate dependent) electrophysiological effects of R56865 (0.48 mg.kg-1) were examined and compared with its antiarrhythmic effect on ouabain-induced ventricular tachycardia (n = 10), and ventricular tachycardia occurring within 24 h of occlusion of the left anterior descending artery (n = 8). The experiments were all performed in dogs. RESULTS: In anaesthetised dogs R56865 increased (p < 0.05) the cycle length of the sinus rhythm, the corrected QT duration (+8%) and the effective refractory period (+16%) of the right ventricle. No rate dependency was found. R56865 had no effect on blood pressure, conduction, or refractoriness of the AV node, nor on conduction in the ventricle. In conscious dogs, R56865 did not change the cycle length of the sinus rhythm, but it did increase the QT duration (+5%, p < 0.05). The cycle length of the slower ouabain induced ventricular tachycardias which were terminated by R56865 increased to a greater extent (+55%) than that of the non-suppressible, faster ventricular tachycardias (+16%): 335(SD 30) ms, n = 5 v 285(10) ms, n = 5. The effect of R56865 on ventricular tachycardias 24 h after infarction was considered to be of minor antiarrhythmic importance. CONCLUSIONS: R56865 has (1) class III effects, (2) a partial effect in terminating ouabain induced ventricular tachycardias which is inverse rate dependent, and (3) a weak effect on ventricular tachycardias 24 h after infarction. PMID- 8221804 TI - Interstitial purine metabolites and lactate during regional myocardial hypoxia. AB - OBJECTIVE: Adenosine is a well known vasodilator believed to contribute to metabolic adjustments of the coronary circulation. The purpose of this study was to assess changes in interstitial fluid adenosine, adenosine metabolites, and lactate during prolonged regional, non-ischaemic myocardial hypoxia. METHODS: To induce regional hypoxia, the left anterior descending coronary artery of anaesthetised dogs (n = 9) was perfused at constant pressure (100 mm Hg) with deoxygenated blood (PO2 approximately 2.6 kPa) for 60 min via an extracorporeal shunt. Cardiac interstitial fluid was sampled by cardiac microdialysis, using dialysate metabolite levels as indices of interstitial fluid concentrations. RESULTS: During hypoxia, coronary blood flow increased 3.9-fold, while myocardial oxygen consumption was maintained relatively constant. There were no changes in global cardiac function, systemic arterial pressure, or heart rate during regional hypoxia, indicating that the hypoxic stimulus did not augment sympathetic nervous system activity. Dialysate adenosine was not increased at any point of the hypoxic period, but was decreased by 25 min hypoxia. Dialysate levels of inosine, hypoxanthine, and xanthine were increased transiently during the first 10 min of hypoxia while there was a sustained increase in dialysate lactate. In the presence of erythro-9-(2-hydroxy-3-nonyl) adenine, an adenosine deaminase inhibitor, adenosine was the predominant purine metabolite and increased transiently during hypoxia. CONCLUSIONS: Flux through the adenosine production and degradation pathways is transiently increased during hypoxia. However, the lack of an increase in interstitial fluid adenosine does not support a role for adenosine in the sustained hyperaemic response to regional myocardial hypoxia. PMID- 8221805 TI - Effect of reperfusion on 111In-antimyosin monoclonal antibody uptake by salvaged and necrotic myocardium in the dog. AB - OBJECTIVE: The aim was to investigate the ability of 111In-antimyosin monoclonal antibody (111In-AMA) to differentiate between salvaged and necrotic myocardium following reperfusion. METHODS: Dogs submitted to a 24 h left anterior descending coronary artery occlusion (group 1, n = 10) or to a 90 min occlusion followed by a 22.5 h reperfusion (group 2, n = 11, group 3, n = 5) were given radiolabelled microspheres and 111In-AMA after 75 min of ischaemia (groups 1 and 2), or after 19.5 h of reperfusion (group 3). After delimiting the area at risk and the infarct by dye perfusion and triphenyltetrazolium chloride, the heart slices were imaged by scintigraphy and dissected into necrotic, viable ischaemic, and normal myocardium. Myocardial blood flow was estimated by microspheres and 111In-AMA uptake was expressed as the ratio of the corresponding non-ischaemic tissue samples taken from opposite ventricular wall. RESULTS: 111In-AMA ratios in necrotic and salvaged myocardium were respectively 5.4(SEM 1.9) and 3.2(0.5) times the normal value, giving a 1.7 to 1 factor between the two areas in dogs with permanent occlusion (group 1). Similar results were obtained in group 3 with ratios of 6.1(1.1) and 3.0(0.3) times normal values. In contrast, ratios of 43.6(5.6) and 5.6(0.9) (p < 0.05) in necrotic and salvaged myocardium, respectively, were found in reperfused group 2, giving a 7.8 to 1 factor between the two tissue areas of the risk territory. Clear delineation between salvaged and necrotic tissue territories could be made on scintigrams only in group 2, which otherwise presented smaller infarcts: 35.1(7.9)% of the risk area v 58.0(8.7)% in non-reperfused animals (p < 0.05). 111In-AMA uptake by necrotic myocardium did not correlate with collateral (group 1) or reperfusion blood flows (group 3), indicating that the greater uptake in reperfused myocardium is flow independent. CONCLUSIONS: 111In-AMA does not clearly identify necrotic from viable ischaemic myocardium within 24 h of injection in a coronary artery occlusion model. Thus it may not be a sensitive enough method to evaluate infarct size progression. However, reperfusion greatly increased 111In-AMA uptake by the infarct in a flow independent manner, this may prove to be useful for clinical assessment of infarct size and reperfusion injury. PMID- 8221806 TI - A model of the time course of myocardial dynamics: use in characterisation of relaxation and evaluation of its indices. AB - OBJECTIVE: Study of myocardial relaxation (lusitropy) requires a lusitropic index that should reflect changes in the time course of relaxation, and is therefore ideally based on a model that encompasses the entire time course of relaxation. Since there is no such model, investigators could not evaluate and validate current measures of relaxation. A new model is therefore proposed and analysed here: X(t) = C(t/A)B(e)1-(t/A)B. X(t) is the time course of isometric force or isotonic shortening, t is time, and A, B and C are parameters. METHODS: The model was analysed and evaluated in nine in situ canine papillary muscles studied with a servo system which produced isometric and isotonic twitches. A, B and C were determined by Marquardt's algorithm. RESULTS: The new model is analytically sufficiently tractable to ascertain, for the first time, whether previous measures of relaxation reflect the time course of relaxation. It nonetheless fits observed data closely, the coefficients of determination being 0.995(SD 0.005) and 0.990(0.004) in isometric and isotonic switches. A and B showed little or no change, with alteration of preload and afterload states averaging 0.200(0.014) s and 0.21(0.022) s for A, and 1.87(0.099) and 2.15(0.158) for B in isometric and isotonic twitches respectively. Dobutamine decreased A but the change was significant only in isotonic twitches; it increased B significantly only in isometric twitches. Thus the lusitropic and chronotropic effects of dobutamine may depend on loading conditions. CONCLUSIONS: This model is uniquely useful because it permits the evaluation and reconciliation of previously proposed measures of myocardial relaxation. It also provides a convenient model of the time course of the mechanical activity of the myocardium under isometric and isotonic conditions and is successful in assigning physiological relevance to its parameters. Moreover it permits expeditious derivation of relations for its parameters, thus providing initial parameter estimates systematically. PMID- 8221807 TI - Attenuation of postischaemic dysfunction by ischaemic preconditioning is not mediated by adenosine in the isolated rat heart. AB - OBJECTIVE: The aim was to test the hypothesis that adenosine mediates the cardioprotective effects of ischaemic preconditioning in the isolated rat heart. METHODS: Transient exposure of the hearts to adenosine and the A1 selective agonist, PIA, were tested for the ability to mimic the cardioprotective effects of ischaemic preconditioning in hearts that underwent 40 min normothermic ischaemic followed by 30 min reperfusion. Treated hearts were perfused with 10 or 50 microM adenosine or 10(-7) M R-phenylisopropyladenosine (PIA) for 5 min followed by a 5 min washout period. Preconditioned hearts underwent 5 min of ischaemia and 5 min of reflow prior to the 40 min ischaemic period. The ability of the adenosine receptor antagonist, BW A1433U, to inhibit the cardioprotective effects of ischaemic preconditioning was also tested. The effects of these treatments on metabolite levels and postischaemic haemodynamic function were assessed. RESULTS: Adenosine (50 microM), but not PIA, resulted in enhanced accumulation of lactate after 40 min ischaemia: 122(SEM 8) v 96(5) nmol.mg-1 protein in control hearts (p < 0.002). Adenosine and PIA treatments did not significantly affect myocardial acidosis during ischaemia. Postischaemic contractile function (as assessed by percent recovery of the heart rate x developed pressure) was lower in 50 microM, but not 10 microM, adenosine treated hearts [8.8(2.2)] and PIA treated hearts [11.9(2.5)] than in control hearts [20.4(3.6)] (p < 0.01). Ischaemic preconditioning (1) lowered glycogen levels prior to the 40 min ischaemic period [57(6) v 110(18) nmol glucosyl units.mg-1 protein; p < 0.01]; (2) lowered lactate levels at the end of the 40 min ischaemic period [61(4) v 104(5) nmol.mg-1 protein]; (3) preserved myocardial pH during ischaemia [6.69(0.07) v 6.40(0.07); p < 0.01]; and (4) enhanced recovery of postischaemic contractile function [42.3(4.4)% v 19.7(6.0)%; p < 0.02]. BW A1433U did not prevent these effects of ischaemic preconditioning. CONCLUSIONS: The cardioprotective effects of ischaemic preconditioning are not mediated by adenosine released during the preconditioning period in the isolated rat heart. Also, transient treatment of the heart with A1 adenosine receptor agonists can exacerbate postischaemic contractile dysfunction. PMID- 8221808 TI - Effect of acute systemic hypoxia on vascular permeability and leucocyte adherence in the anaesthetised rat. AB - OBJECTIVE: The aim was to investigate the effect of acute systemic hypoxia on vascular permeability to macromolecules and on leucocyte adherence to vascular endothelium in vivo. METHODS: Experiments were performed on anaesthetised rats with either the intestinal mesentery or the spinotrapezius muscle prepared for in vivo microscopy. To quantify changes in vascular permeability, fluorescein isothiocyanate conjugated with serum albumin (FITC-albumin) was given intravenously and the microcirculation was viewed using a mercury source for 30 s periods during air breathing; or before, during, and after breathing 6% O2 for 3 or 20 min. On each occasion the number of FITC leakage sites was counted. In separate experiments acridine orange was given to stain leucocytes and the microcirculation was viewed using a mercury source during air breathing and during a 3 min period of systemic hypoxia. The number of leucocytes that adhered to venular walls for > 30 s was counted. Using mesentery, the effects were tested of BW755C, a lipoxygenase inhibitor, and of SM9064, a LTB4 receptor antagonist, upon the increase in leucocyte adherence observed during hypoxia. RESULTS: In rats that breathed air throughout, the number of leakage sites for FITC-albumin in both the spinotrapezius and mesentery remained constant. Moreover, in rats that breathed 6% O2 for 3 or 20 min, the number of leakage sites was not changed in either mesentery or spinotrapezius by hypoxia, but was substantially increased in both preparations by topical application of histamine. However, the number of leucocytes that adhered to the inside of venular walls was significantly increased in both mesentery and spinotrapezius by a 3 min inhalation 6% O2 from 2.83(SEM 0.56) to 4.66(1.77) per 100 microns length of venule and from 2.44(0.33) to 3.35(0.49) respectively during the first period of hypoxia. Between periods of hypoxia the number of adherent leucocytes returned to control in both preparations. Leucocyte adherence was not affected by BW755C (50 or 500 micrograms.ml-1 applied topically or 10 mg.kg-1 intravenously) or by SM9064 (3 mg.kg-1 intravenously). CONCLUSIONS: Acute systemic hypoxia does not affect the vascular permeability to albumin. However, 3 min periods of systemic hypoxia induce significant, but reversible, increases in leucocyte adherence in both muscle and mesenteric venules which in mesentery, at least, is not mediated by LTB4 or other products of the lipoxygenase pathway. PMID- 8221809 TI - Studies of cardiopulmonary bypass in children: implications for the regulation of brain natriuretic peptide. AB - OBJECTIVE: The aim was to examine the influence of cardiopulmonary bypass on brain natriuretic peptide (BNP) and on hormones of importance in the control of sodium and water balance and blood volume. METHODS: Nine patients (mean age 4 years, range 2-9) undergoing cardiac surgery were studied. Blood samples were taken before, during, and up to 24 h after bypass. Plasma levels of BNP, atrial natriuretic peptide (ANP), arginine vasopressin (AVP), plasma renin activity, aldosterone, and catecholamines were measured. RESULTS: Preoperative concentrations of plasma BNP [573(SEM 68) pg.ml-1] and ANP [332(74) pg.ml-1] were greatly increased (p < 0.05) before bypass in all patients when compared to normal levels in children [BNP = 31(4) pg.ml-1; ANP = 27(3) pg.ml-1, n = 28]. With general anaesthetic and sternotomy, there were large reductions (p < 0.05) in both plasma BNP [180(62) pg.ml-1] and plasma ANP [163(59) pg.ml-1]. During bypass, there were no further significant decreases in plasma ANP or BNP concentrations compared with preoperative levels. Postoperatively, plasma BNP gradually increased for 12 h, to 170(28) pg.ml-1, whereas plasma ANP showed a further small decrease, to 107(20) pg.ml-1. However, postoperative plasma levels of both ANP and BNP remained well below preoperative values (p < 0.01). Plasma AVP increased rapidly within 15 min of the onset of bypass, reaching a peak value of 153(5) pg.ml-1 after 45 min. Off bypass, plasma AVP decreased slowly and was still almost 10-fold above preoperative levels 12 h after end of bypass [137(11) pg.ml-1]. Mean central venous pressure decreased during the onset of bypass, from 4.3(1.9) to 0.4(1.1) mm Hg (p < 0.05), and increased again at the end of bypass, to 9.0(3.3) mm Hg (p < 0.05); there was little further change during the postoperative period. CONCLUSIONS: The major source of plasma BNP in patients with congenital heart disease is the cardiac ventricle. The lower plasma ANP and BNP levels and the narrow band of change in central venous pressure following surgical repair of cardiac abnormalities may be a response to improved cardiac function. PMID- 8221810 TI - Downregulation of oxygen demand in isoprenaline stimulated canine myocardium. AB - OBJECTIVE: The aim was to evaluate the hypothesis that the oxygen demand of stimulated myocardium is downregulated and ischaemia avoided when oxygen supply is limited. METHODS: Isoprenaline stimulation during reduced coronary perfusion pressure or moderate coronary hypoxaemia was studied in relation to myocardial oxygen demand, power, and oxygen utilisation efficiency (O2Effic) in 19 anaesthetised open chest dogs. The left anterior descending coronary artery was perfused with normoxic blood at 100 or 60 mm Hg or with moderately hypoxic blood [CaO2 = 9.8(SEM 0.2) ml.dl-1] at 100 mm Hg. Myocardial oxygen demand was estimated from oxygen consumption (MVO2), and power index was computed from heart rate, left ventricular pressure, and segment shortening. O2Effic was calculated from power index divided by MVO2. Lactate extraction and contractile function were used as indices of myocardial ischaemia. RESULTS: Isoprenaline under control conditions increased coronary blood flow, MVO2, and power index, but did not alter lactate extraction. Reducing coronary perfusion pressure to 60 mm Hg decreased oxygen delivery by 31.6(5.7)% (p < 0.05). MVO2 fell by 26.6(8.6)% (p < 0.05), but power index and lactate extraction were not reduced. At reduced coronary perfusion pressure, isoprenaline-induced elevation of MVO2 decreased from 26.0(1.5) to 15.7(2.6) ml.min-1 x 100 g-1 (p < 0.05), but the isoprenaline induced rise in power index was not significantly lessened, nor was lactate extraction altered. O2Effic was increased by reduced coronary perfusion pressure and by isoprenaline during reduced coronary perfusion pressure. Hypoxaemia increased coronary blood flow but did not alter oxygen delivery, MVO2 or power index. Isoprenaline-induced elevation of MVO2 decreased from 25.2(1.7) to 19.1(0.5) ml.min-1 x 100 g-1 (p < 0.05), and power index decreased by 33(9.1)% (p < 0.05). O2Effic was not altered by hypoxaemia or by isoprenaline during hypoxaemia. CONCLUSIONS: Oxygen demand of isoprenaline stimulated myocardium is downregulated and ischaemia is avoided when oxygen is limited. Downregulation of oxygen demand is achieved by increasing oxygen utilisation efficiency in the presence of reduced coronary perfusion pressure and by decreasing power in the presence of hypoxaemia. PMID- 8221812 TI - A numerical method for the calculation of the time constant of exponential functions. PMID- 8221811 TI - The effect of cytokine(l-n) on vascular/myocardial function. PMID- 8221813 TI - Treating hypertension in the diabetic patient: therapeutic goals and the role of calcium channel blockers. AB - Although many drugs effectively control hypertension in patients with diabetes mellitus, the risk of aggravating the diabetic process and inducing new complications can negate the benefits of reduced blood pressure. This article discusses the current status of antihypertensive therapy in diabetic patients and focuses on the use of calcium channel blockers that safely reduce blood pressure while having a neutral or beneficial effect on the diabetic state and related medical complications. While both calcium channel blockers and angiotensin converting enzyme (ACE) inhibitors are suitable agents for the primary treatment of diabetic hypertension, the beneficial effects of calcium channel blockers on glucose homeostasis, lipid metabolism, and renal function support their routine use as alternatives to ACE inhibitors. Among the three classes of calcium channel blockers, second-generation dihydropyridines offer advantages over older calcium channel blockers because the second-generation drugs provide greater vascular selectivity and less direct myocardial depression. PMID- 8221814 TI - Ambulatory blood pressure measurement in assessing the antihypertensive effect of benazepril plus hydrochlorothiazide in a fixed combination. AB - Ambulatory 24-hour blood pressure monitoring was used to assess the antihypertensive efficacy of the angiotensin-converting enzyme (ACE) inhibitor benazepril in combination with the diuretic hydrochlorothiazide (HCTZ) in 17 mildly to moderately hypertensive patients. Compared with placebo, benazepril 10 mg + HCTZ 12.5 mg induced a statistically and clinically significant reduction in the mean, systolic, and diastolic blood pressures. Cumulative percentages of diastolic blood pressure readings < 90 mmHg were also greater with the combination treatment than with placebo. No patient discontinued the treatment because of adverse effects. PMID- 8221815 TI - Six-month maintenance treatment of duodenal ulcer with sucralfate: influence on antral gastritis and Helicobacter pylori antral colonization--a prospective uncontrolled study. AB - Fifty-four patients (41 men, 13 women), aged 17 to 78 years (mean +/- SD, 48.13 +/- 13.5 years), with endoscopically confirmed healing of their duodenal ulcer after treatment with sucralfate (2 gm BID for 4 to 8 weeks) were recruited for this study. They were started on a 6-month maintenance treatment with sucralfate 1 gm BID. Endoscopy was done at the end of the 6-month period or whenever there was any evidence of ulcer relapse. Helicobacter pylori antral colonization (CLO test) and antral gastritis were estimated from biopsy samples taken before, and at the end of, the healing treatment, as well as at the end of the maintenance treatment. Cumulative relapse rate after 6 months was 15% (8 of 54). No patient discontinued treatment because of side effects. No influence of sucralfate on H pylori antral colonization or antral gastritis was observed after the healing or maintenance treatment. It is concluded that sucralfate 1 gm BID for 6 months is an effective maintenance treatment for duodenal ulcer, but has no beneficial effect on either H pylori antral colonization or antral gastritis. PMID- 8221816 TI - Chronic or recurrent vaginal candidosis: short-term treatment and prophylaxis with itraconazole. AB - Twenty-one women (mean age, 35.3 years) with 2 to 4 episodes of vaginal candidosis in the last 6 months were included in the study to evaluate the efficacy and safety of itraconazole as short-term, as well as prophylactic, treatment of chronic candidosis. After clinical evaluation and laboratory confirmation of candidosis, 200 mg of itraconazole were given orally for 3 days. Twelve of the 21 patients were cured. The remaining nine repeated treatment, after which all were cured. All patients were entered in the maintenance phase, and received 200 mg of itraconazole the first day of the menstrual cycle for 6 months. One patient relapsed in the second month, but after taking 200 mg of itraconazole BID for 1 day she remained cured for the rest of the study period. All other patients remained cured for the 6 months of the maintenance period. Three months after the end of prophylactic therapy, 17 of 20 patients (85%) were clinically and mycologically cured. No adverse experiences were reported. It is concluded that itraconazole is an efficient and safe short-term treatment for chronic or recurrent vaginal candidosis. Moreover, the dose of 200 mg once monthly for 6 months proved to be a successful suppressive treatment. PMID- 8221817 TI - Hepatic arterial infusions of interleukin-2-based immunochemotherapy in the treatment of unresectable liver metastases from colorectal cancer. AB - In preclinical studies, hepatic arterial infusion of interleukin-2 (IL-2) in dogs significantly induced lymphocyte proliferation and augmented antitumor killing activity in the liver. Based on these findings, a pilot study of hepatic arterial infusions of IL-2-based immunochemotherapy was conducted in 21 patients (15 men, 6 women) with unresectable liver metastases from colorectal cancer, to determine whether the addition of IL-2 improved the therapeutic efficacy of chemotherapy alone. Interleukin-2 was given to all patients as 7 to 8 x 10(5) Japanese reference units (JRU) in addition to 5-fluorouracil (5-FU) 250 mg daily and mitomycin C (MMC) 4 mg once weekly, through a subcutaneous port for 3 weeks. After completion of the initial course, patients were discharged from the hospital and continued on a modified regimen for outpatient therapy: IL-2, 2.0 to 2.1 x 10(6) JRU and 5-FU 250 mg twice weekly; MMC 4 mg once weekly. Patient response rate was 76%, and the median survival from initiation of treatment was 24 months. Toxicity of the combined regimen was minimal. Peripheral lymphocyte phenotype study showed notable decreases in CD8+, CD16+, and CD57+ cells and an increase in CD4+ cells (ie, elevation of 4:8 ratio) during therapy. Electron microscopic analysis of the resected liver of a patient receiving the IL-2 mitomycin-C/5-fluorouracil (IL-2.MF) infusion showed a pronounced accumulation of lymphocytes, penetrating from the space of Disse, around the cancer cells. The present study explores hepatic arterial infusion of IL-2-based immunochemotherapy as a new strategy, based on the activation of liver-associated immune response; this technique may provide improved response and survival for unresectable liver metastases. PMID- 8221818 TI - Pilot study of a new angiotensin II receptor antagonist, TCV-116: effects of a single oral dose on blood pressure in patients with essential hypertension. AB - After a 1-week placebo control period, six hypertensive patients (mean age, 67 years) each received single doses of 1, 2.5, and 5 mg of TCV-116 at 2- to 3- day intervals. Systolic and diastolic blood pressures were significantly lower after the final dose (5 mg) of TCV-116 than on the last day of the placebo period. Blood pressures were decreased after each dose of TCV-116 in a dose-dependent fashion from 2 hours after administration and reached a nadir at 4 to 6 hours. After 2.5 and 5 mg of TCV-116, the hypotensive effect was sustained for 24 hours. Pulse rate did not change significantly. Plasma renin activity and angiotensin I levels increased in a dose-dependent fashion after TCV-116, but the changes were not significant. No changes were noted in plasma aldosterone or angiotensin II levels. One patient reported mild light-headedness after 5 mg of TCV-116. No other side effects or abnormal laboratory tests results were noted. It appears that TCV-116 is a safe and effective antihypertensive agent. PMID- 8221819 TI - Effects of enalapril maleate on heart rate variability: a pilot study. AB - The effects of enalapril maleate on heart rate variability, an indicator of cardiac autonomic function, were evaluated by nonspectral analysis in 12 patients with standard deviation in the R-R interval for each 5-minute period for 24 hours (SDANN) of 80 msec or less. A 24-hour electrocardiogram was recorded before treatment and after 2 weeks of treatment with enalapril. No significant difference was observed in the mean R-R interval on 24-hour electrocardiography between the pretreatment and the posttreatment periods (824 +/- 84 msec vs 834 +/ 83 msec). The SDANN increased significantly (P < 0.01) between the pretreatment period (62.8 +/- 13.7 msec) and the posttreatment period (87.1 +/- 31.2 msec), as did the standard deviation of the R-R interval during the two monitoring periods (P < 0.001). However, no significant difference was observed in the mean value of the standard deviations of the R-R interval for 5 minutes; the proportion of adjacent R-Rs with a difference of more than 50 msec; or the root-mean square of the difference in successive R-Rs in the pretreatment period as compared with the posttreatment period. Causal blood pressure at rest decreased significantly (P < 0.05) after treatment with enalapril (124 +/- 17/66 +/- 7 mmHg) as compared with the pretreatment period (140 +/- 24/75 +/- 13 mmHg). Enalapril may be useful for maintaining normal autonomic nerve activity, preventing sudden cardiac death, and enhancing quality of life in our stressful society. PMID- 8221820 TI - Amino acids for dialysis patients. AB - Calorie/protein malnutrition is a common problem among dialysis patients. A study was designed to determine the value of adding a mixture of amino acids to the diet of 36 patients (mean age, 42 years) who had been receiving triweekly hemodialysis for periods ranging from 1 to 13 years. For 1 month the patients received a mixture containing 8 essential and 9 nonessential amino acids. Before treatment, the mean levels of most amino acids were lower in the patients than in a control group of 18 healthy subjects (mean age, 43.5 years). Significant increases in most amino acids were noted in the patients after treatment, including marked increases in tryptophan, valine, leucine, isoleucine, lysine, threonine, taurine, aspartic acid, serine, histidine, tyrosine, and alanine. The essential:non-essential and tyrosine:phenylalanine ratios increased significantly during treatment. Meanwhile, serum hematocrit levels increased and phosphorus and parathormone levels decreased significantly; no changes were noted in urea, creatinine, or total protein levels. The patients reported an improvement in health and in tone and mood after treatment. The results confirm the value of dietary supplementation for dialysis patients. PMID- 8221821 TI - Effects of sustained-release isradipine on blood pressure and peripheral hemodynamics in hypertensive patients. AB - In a double-blind crossover study, 16 hypertensive patients (mean age, 41 years) were randomly assigned to receive placebo or 5 mg of an extended-release formulation of isradipine for 30 days. Blood pressure and heart rate were recorded by an automatic device and hemodynamics measured by a duplex scanner and plethysmography. After the first dose and after 30 days' treatment with isradipine, blood pressure was significantly reduced (mean arterial pressure 4 hours after the first dose, 106 +/- 3 vs 120 +/- 4 mmHg, P < 0.01; 22 hours after the last dose, 108 +/- 3 mmHg, P < 0.01) with no significant changes in heart rate. The compliance of the brachial artery was significantly increased (2.823 +/ 0.358 vs 1.204 +/- 0.156 dyn-1.cm4.10(-7), P < 0.002) and the characteristic impedence decreased (49 +/- 6 vs 91 +/- 12 dyn.s.cm-5.10(2), P < 0.05) as well as local resistances (71 +/- 5.6 vs 198 +/- 18 mmHg.ml-1.s, P < 0.001). After 30 days of isradipine treatment, 22 hours after the last dose, compliance was still increased (2.575 +/- 0.453 dyn-1.cm4.10(-7), P < 0.01) whereas impedance and forearm vascular resistances were reduced (59 +/- 8 dyn.s.cm-5.10(2), P < 0.05, and 97 +/- 14 mmHg.ml-1.s, P < 0.001, respectively). The results indicate that sustained-release isradipine ensures good blood pressure control up to the time of the following dose and restores the large artery dumping function against cyclic variations in intraluminal pressure. PMID- 8221822 TI - Effect of pharmaceutical formulation for antihypertensive therapy on health service utilization. AB - A significant factor in the management of hypertension is the extent to which patients comply with the treatment regimen. A retrospective analysis was undertaken to determine the relationship between antihypertensive formulation, regimen compliance, and the utilization of health care services. Data for this analysis were derived from the state of South Carolina's Medicaid computer archive. The study population consisted of 1000 randomly selected patients initially prescribed one of the following antihypertensive regimens as monotherapy: atenolol once daily, captopril BID, oral clonidine BID, transdermal clonidine once weekly, diltiazem BID, enalapril BID, metoprolol BID, prazosin BID, terazosin once daily, and sustained-release verapamil once daily. Multivariate regression analysis was used to determine the incremental influence of selected demographic characteristics, use of medical services before diagnosis of hypertension, initial antihypertensive medication, medication possession ratio for antihypertensive therapy, and number of maintenance medications for diseases other than hypertension on post-period health care expenditures. The results indicated that patients initially prescribed antihypertensive medication requiring once-daily or once-weekly administration experienced an increased utilization of antihypertensive medication, needed fewer changes in their therapeutic regimen, and far less need for concomitant therapy for blood pressure control compared with those prescribed a BID regimen. Patients in the once-daily or once-weekly groups also used significantly fewer physician, hospital, and laboratory services (P < or = 0.05). PMID- 8221823 TI - Points to consider about prescription drug prices: an overview of federal policy and pricing studies. AB - Pharmaceutical prices in the United States are under evaluation as policy makers decide how to reduce health care expenditures and public subsidy of the pharmaceutical industry. Furthermore, evidence of higher drug prices in the United States, compared with those in other countries, fuels the prescription drug price debate. These issues are not new to the public forum and much can be learned from prior debates and policies. This paper begins by reviewing the pricing debate with the Kefauver hearings on monopolies held during the late 1950s and early 1960s and continues with the current price debate. Government reports and academic studies are discussed, addressing the methodological differences and their implications to policy makers. Finally, the literature review includes foreign government prescription drug programs with their respective prescription drug expenditures. Evidence provided by academics about the pricing practices of the drug manufacturers indicates product quality and price information would force firms to compete on the price level, thus reducing pharmaceutical product pricing to the "true" market price. PMID- 8221824 TI - The political economy of pharmaceutical marketing channels: a conceptual framework. AB - The political economy framework is presented as a guide to investigate pharmaceutical marketing channels. Based on the framework, pharmaceutical marketing channels are conceptualized as four interacting areas: the environment, the economy, the policy, and the performance of the channel. Examples of this approach are illustrated. The usefulness of the framework for future research and management of pharmaceutical marketing channels is discussed, and strengths and weaknesses are identified. PMID- 8221825 TI - The nature and consequences of policies intended to contain costs in outpatient drug insurance programs. AB - Outpatient drug insurance programs play an important role in making drugs affordable. Public and private program policy-makers often respond to fiscal pressures by reducing benefit coverage or pharmacy reimbursement levels. These reductions defeat both coverage and cost-containment goals. A more prudent approach is to adopt a comprehensive policy for outpatient drug coverage that protects patients and ensures fair and prompt reimbursements to pharmacy providers. These issues are well illustrated by the economic situation in Illinois, resultant state budget problems, and the consequences for the Illinois Medicaid outpatient drug program and the Illinois state-only pharmaceutical assistance program for low-income elderly patients. The questions examined in this paper have important implications for patients, pharmacists, physicians, the pharmaceutical industry, public and private policymakers, and society as a whole. PMID- 8221826 TI - [Health policy in the Czech Republic]. AB - Based on an analysis of the development of the health status of the population of different European countries during the last 30 years of the authors demonstrate the dominant determining role of the character of the social system for this development. In conjunction with actual situation and the transformation of the entire society the authors outline basic socioeconomic prerequisites for the creation of a positive health policy of the Czech Republic. Objectives and the main strategic steps for their implementation are outlined. PMID- 8221827 TI - [The direction and mission of social medicine]. AB - The author characterizes briefly the contemporary baseline situation of social medicine. He explains its three aspects--the methodical and cognitive approach, the orientation on values and attitudes and the formative and action component. The author emphasizes the integral concept and the necessity of a purposeful development of the mentioned discipline. The purpose of the article is to provide basic orientation in social medicine for discussion and creative work. PMID- 8221828 TI - [A demographic model of hospitalization rates in the population of the Czech Republic as a tool for evaluation of inter-district differences]. AB - The authors elaborated a computer model of the hospitalization rate of the population, using the method of indirect standardization which can apply the available specific hospitalization rate of a major territorial unit by age and sex to defined regional sub-populations. The model was used to the calculation of the theoretical standardized hospitalization rate in hospitals and sanatoria for chronic patients in Prague and different district of the Czech Republic, using the specific hospitalization rate of the whole Czech Republic as a standard for 1986 to be compared with the actual number of patients hospitalized in that year. The ratio of the actual number of hospitalized patients and the calculated standard for 1986 varied between 73.5% in the district Plzen-north and 125.9% in the Bruntal district. In the Bruntal district the age standardized hospitalization rate was thus by more than 70% higher than in the Plzen-north. In district with the highest ratio of the actual and standardized number of hospitalized patients there was in all instances a relatively great excess of beds which was obviously the main subjective reason for the high hospitalization rate of the population. In the district of Cheb, Svitavy, Most, Chomutov and Bruntal there were, as compared with the Czech average of 400 almost some 600 beds in hospitals and sanatoria for chronic patients in excess. The hospitalization rate of the population in the Czech Republic in 1986 is compared with that in Holland in 1989-1991 and it was found that the comparable Czech data were by 1.8 times higher. PMID- 8221829 TI - [Life style of medical students and risk factors for ischemic heart disease]. AB - For three years some characteristics of life-style of medical students in the fifth year of The 1st Medical Faculty, Charles University, Prague, were followed with a view to risk factors of ischemic heart disease (ICHS). At the same time opinion of these students on the validity of these factors were established. Altogether it deals with 623 students of which they are 269 men and 354 women of average age 23.1 years in whom 12 risk factors were followed. The results show that medical students do not lead an exemplary life-style. For example, during the following three years there was actually a significant increase in number of smokers (from 7% to 22%) and corresponding to this the significance assigned to this risk factor has decreased. PMID- 8221830 TI - [Ethical decision-making on limitations and needs in terminal care]. AB - Many physicians are refusing euthanasia as a possible solution of a patient's excessive distress, being aware of their responsibility for decision-making in the terminal care. A sharp dividing line has to be drawn between accepting inevitable death and unjustifiable manipulations with human life based on arbitrary qualitative criteria. Ethical decision-making should proceed from the particular clinical situation, which influences the definition of ordinary and extraordinary therapeutic means. Contemporary medicine has progressed towards authentic terminal care, respecting the fundamental needs of the terminally ill patient, in all dimensions of human person. PMID- 8221831 TI - [Medicine and music]. PMID- 8221832 TI - [An analysis of the risk factors of child bronchial asthma]. AB - We have analyzed the risk factors of child bronchial asthma by a 1:1 matched case control study. The results showed that the family hereditary history (OR = 44.44); the history of ingesting sensitizing foods (OR = 22.52) and of allergy (OR = 7.11) in maternal pregnancy; the history of allergy in childhood (OR = 9.27) and of respiratory diseases in infancy (OR = 7.71); the personality of child (OR = 8.01), the residential environment (OR = 9.08), and so on were the risk factors of child bronchial asthma. In addition, we have found that the age of the 1st onset of child bronchial asthma was associated with the following risk factors: the history of allergy in childhood and in maternal pregnancy, the family hereditary history and the residential environment, etc. It is confirmed that the child bronchial asthma is a multifactor disease. PMID- 8221833 TI - [A study on the carrier state of Haemophilus influenzae in pharynx in healthy children in Beijing Area]. AB - One hundred and fifteen healthy children in 2 kindergartens of Beijing City were enrolled in a study for carrier state of Haemophilus influenzae (HI) in pharynx. Nontypable HI flora was found in 32.2% (37/115) in this group. There were no significant differences of carrier state in sex, age, and season in these children. Antibiotic sensitivity test showed that some HI strains isolated were resistant to ampicillin and tetracycline (5/34 vs 4/34), most strains resistant to SMZ Co (21/34). PMID- 8221834 TI - [Establishment and application of quality assessment program for HIV testing laboratories]. AB - For helping HIV testing laboratories to improve their testing quality, a national external quality assessment (QA) Program was established and 2 QA survey were carried out in 1991 and 1992. It suggested that the established QA program, especially the scoring system, was suitable for use in our country and could be used to evaluate the technical level of testing laboratories. The results of QA showed that the techniques of the regional HIV testing laboratories had reached a rather high level and could perform screening and confirmatory tests correctly. PMID- 8221835 TI - [An investigation on traffic accidents in Shanghai from 1986 to 1990]. AB - During 1986 to 1990, the rate of traffic accidents was 66.69, that of mortality 5.48, and that of injury 44.62 per 100,000 person-year in Shanghai, but the rates decreased since 1987. The accident and mortality rates in 12 urban districts were significantly lower than in 9 rural counties (P < 0.01). Of 1967 accidental deaths, the main responsibility in 39.20% belonged to drivers, and that in 36.81% belonged to the persons riding bicycles. And 32.27% accidental deaths were caused by the drivers from other provinces. PMID- 8221836 TI - [A seroepidemiological investigation on HCV infection among population in Tianjin area]. AB - Sera from 88 hepatitis B (HB) patients and 375 cases with gynecological conditions were detected for anti-HCV antibody with ELISA. The positive rate of anti-HCV antibody in HB cases was 12.50% (11/88); in gynecological cases was 0.80% (13/375). It was confirmed that HCV infection not only existed in this area but was also associated with the type of HB. The results suggested that HCV infection might be an important factor for HB to become chronic and difficult to treat. The positive rate of anti-HCV antibody in gynecological cases was quite high, suggesting that nosocomial infection should be paid attention to. In addition to the strict disinfection of medical equipment and blood products, the screening of blood donors, defining the indications for transfusion of blood and blood products and immunizing the population at high risk with immunoglobulin were recommended. PMID- 8221837 TI - [Thermostability and immunogenicity of trivalent oral poliovirus vaccine produced by different countries]. AB - Hep-2 cell-based microtiter assay system was used to test the titer of TOPV(Sabin strain). Before immunization, the overall potency results of TOPV produced by China, Yugoslavia, and Belgium were 6.38, 6.25, and 5.75 log TCID 50/0.1 ml, respectively. After storage at 37 degrees C for 2 days or after 5 cycles of freezing and thawing, the overall titer of each TOPV reduced less than 0.5 log TCID 50/0.1 ml. A randomized study was conducted in Anhui Province to test the serological difference among the TOPV which were produced by China, Yugoslavia and Belgium. The formulations of the TOPV produced by China and Yugoslavia were 6.0, 5.0, and 5.5 log TCID 50/0.1 ml of Sabin types 1, 2, and 3, respectively. All of the children in this study were divided into three groups, and children in group A, B, and C received TOPV produced by China, Yugoslavia, and Belgium, respectively. Three doses of TOPV were given to the children in the group at 2, 3, and 4 months of age. The seroconversion rates and geometric mean titers had no significant differences among these groups before immunization. At the end of one month following the 2nd dose, the rates of seroconversion to type 1-3 poliovirus in group A, B, and C were 88%, 98%, 98%; 92%, 98%, 92%; 100%, 100% and 96%, respectively. The rate of seroconversion to type 1 poliovirus in group C was higher than those in group A and B.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8221838 TI - [An observation on immunization reaction and serological effect of adsorbed purified pertussis vaccine]. AB - An observation on immunization reaction and serological effect of adsorbed purified pertussis vaccine for the first time in China had been carried out in Zhengzhou. Phase I field trial had been conducted in 8 adults and 20 children in whom there was no local reaction and temperature reaction in adults and only mild temperature reaction (37.6 degrees C-38 degrees C) in 4 children. Phase II field trial showed that the adsorbed purified pertussis vaccine manifested trivial reaction, none developed severe temperature reaction (39.1 degrees C), and 8 person-times of moderate temperature reaction (38.1 degrees C-39.1 degrees C) occurred in 100 children after 3 doses. Whereas 2 person-times of severe reaction and 39 person-times of mild reaction occurred in 101 children after receiving the whole cell pertussis vaccine. None showed local severe reaction (5.1 cm) and 1 person-time of mild local reaction appeared at 48 hours after each of 3 doses of adsorbed purified pertussis vaccine (102, 102, 98 children, respectively); Whereas, 3 person-times of severe local reaction and 17 person-times of mild local reaction occurred after each of 3 doses of whole cell pertussis vaccine (97, 100, 97 children, respectively). Satisfactory immunogenicity of the adsorbed purified pertussis vaccine was confirmed. The high serum hemagglutinin titer was achieved in 90 children, the GMT reached 817 (9.16 for preimmunization), and titers reaching 1:320 and over (a level protecting human from being infected) accounted for 84.4 percent. The average levels of anti-LPF and anti-FHA antibodies in 60 and 53 persons attained to 24.17 +/- 3.9421 EU/ml and 38.85 +/- 7.2466 EU/ml, respectively. PMID- 8221839 TI - [A study on risk factors and control of neonatal tetanus during 1975-1990 in Yichang City]. AB - Retrospective survey on the mortality of 21 cases of neonatal tetanus during 1975 1990 and their case-control study on death risk factors were reported in Yichang City. Surveillance and control of neonatal tetanus were emphasized on suburbs. Serological survey of tetanus antibody in women aged 15 to 40 and mothers and their newborn babies were carried out. The protective rate was 3.75% (GMT = 0.0014 IU/ml). The result showed that tetanus antibody in the women of childbearing age was at the lowest level. There was a significant positive correlation through vertical transmission between mothers and their newborn babies (r = 0.7096, P < 0.0005). And the successful immunization in 65 pairs of children was 100%. It was suggested that we should consolidate EPI and promote maternal and child health plan and a primary-health-care program in Yichang City. PMID- 8221840 TI - [Survey and analysis on correlative factors with CHD in 12,468 pupils in Yimeng mountain area]. AB - The risk factors correlated with congenital heart diseases (CHD) in 12,468 pupils in Yimeng mountain area were surveyed and analysed. The morbidity rate of CHD was 3.05%, which was higher than those of other heart diseases; ventricular septal defect (VSD) was more frequent than other types of CHD. The frequency in pupils in city is higher than in rural areas. In 38 cases with CHD, through analysis (1:1) on the risk of single factor and the progressive regression analysis on compound factors, the results showed that the age of pregnant women (> or = 35 years), the job of mother (chemical worker), the history of coryza in pregnancy (within 3 months), the season of birth (August, September), and the family history of malformation are notably correlated with CHD. PMID- 8221842 TI - [Clinical epidemiology]. PMID- 8221841 TI - [A survey on enteroviruses-carrying status among healthy population in Changle and Wucheng counties of Shandong Province. Shandong Collaborative Group of Poliomyelitis]. AB - Four-hundred and fourty-six faecal specimens sampled from healthy carriers between 0 and more than 20 year-old age groups in Changle and Wucheng counties of Shandong Province were collected from June to Sept. of 1991. From them, 142 (31.84%) isolates of enteroviruses were obtained. There was no significant difference in the enteroviruses isolation rate between Changle and Wucheng. No poliovirus was isolated. Among other enterovirus isolates, 85 were echoviruses, 6 coxsackieviruses, 9 enterovirus type 70 and 2 enterovirus type 71. Other 40 isolates were positive for cytopathogenic effect (CPE) in cell culture, but they could not be identified by antisera to poliovirus and enteroviruses. The results showed that the infection by non-polio enteroviruses which could cause polio-like paralysis should be paid attention in the poliomyelitis eradication program. PMID- 8221843 TI - Late outcome and quality of life following percutaneous transluminal coronary angioplasty in octogenarians. AB - Limited data are available concerning the long-term survival and quality of life of octogenarians treated with percutaneous transluminal coronary angioplasty (PTCA). We retrospectively compared the results of PTCA in 118 octogenarians to that of 500 younger subjects. Among the octogenarians, long term follow-up was obtained including survival, relief of angina, assessment of quality of life, and capacity for independent living. These results were compared between men and women. The clinical success rate was 93% for the octogenarians and 88% for the younger patients (p = NS). The lesion success rates were 89% and 88% for these groups, respectively. Results of multilesion PTCA and dilatation of coronary occlusions were similar in both age groups. Major complication rates were 5.9% for the elderly and 3.8% for the younger patients (p < .008). Hospital mortality was higher among the octogenarians (4.6% vs 0.2%, p < .05). Among long-term octogenarian survivors, > 90% indicated a high level of satisfaction with their quality of life and health status. If needed, 88% would undergo the procedure again. Independent living was possible for 66% of these patients and automobile driving for 55%. Octogenarian men and women had similar baseline clinical characteristics, procedural success, complications, relief of angina, and survival. Women indicated a slightly lower level of satisfaction and capacity for independent living. PTCA can achieve a success rate in octogenarians comparable to that of younger patients but with an increased risk of serious complications. Patient satisfaction remains high and a majority of octogenarians remain physically active and capable of independent living. PMID- 8221844 TI - Cardiac catheterization and coronary angiography using 5 French preformed (Judkins) catheters from the percutaneous right brachial approach: a comparative analysis with the femoral approach. AB - This study describes a method for the performance of cardiac catheterization using 5 French preformed Judkins catheters from a percutaneous right brachial approach, and compares that technique to the more traditional percutaneous right femoral approach with 6 French catheters. One hundred consecutive patients requiring diagnostic left heart catheterization and selective coronary angiography were randomized according to femoral versus brachial arterial technique. Procedural efficiency, radiation exposure, and diagnostic film quality favored the femoral approach, while patient comfort, hemostasis time, time to ambulation, and decreased need for post-procedure nursing care favored the brachial approach. No differences were identified in complications. Cardiac catheterization from a right brachial artery percutaneous approach with 5 French preformed catheters has both advantages and disadvantages when compared with a more traditional femoral approach with 6 French catheters. Multiple factors should be considered before selecting an approach to diagnostic cardiac catheterization and each patient should be individually evaluated for determination of the optimal technique. PMID- 8221845 TI - Transcatheter occlusion of the patent ductus arteriosus in the presence of mild isthmal hypoplasia. AB - Between January 1990 and February 1992, 5 asymptomatic patients were identified with hypoplasia of the aortic isthmus at the time of cardiac catheterization for closure of a small patent ductus arteriosus. Isthmal diameters measured from lateral aortograms were 62 +/- 11% of age-matched controls. All patients had a small communication occluded with a 12 mm device and remained asymptomatic at 6 30 months follow up. Residual shunting was demonstrated by color Doppler in 2 patients. No child has developed an arm-leg systolic blood pressure gradient or systemic hypertension. Following device placement, Doppler interrogation across the isthmus in 2 patients demonstrated a velocity shift of 2.1 m/s. Transcatheter occlusion of the patient ductus arteriosus associated with mild isthmal hypoplasia of the aortic arch appears safe and effective and does not result in a clinically significant impairment to isthmal flow in short-term follow-up. PMID- 8221846 TI - Anomalous origin of the left circumflex coronary artery from the right aortic sinus: a familial clustering. AB - Origin of the left circumflex coronary artery from the right sinus of Valsalva or the right coronary artery is a well-described anomaly. We report 3 cases which suggest a familial association of this anomaly. The familial clustering that we report has not been previously demonstrated. PMID- 8221847 TI - Use of angioplasty in the management of complicated perioperative infarction following bypass surgery. AB - Ischemic complications in the perioperative period following bypass surgery adversely affect both short- and long-term prognosis. Coronary angioplasty was successfully performed in 2 patients sustaining complicated postoperative myocardial infarctions with resolution of angina and restoration of hemodynamic stability. These cases illustrate that angioplasty can be done safely in such patients and may favorably alter short-term outcome. PMID- 8221848 TI - Nineteen years survival after occlusion of the left main coronary artery by virtue of an anomalous septal branch. AB - This case presents a patient who survived 19 years with an occluded left main coronary artery as a result of collateral flow to the anterior wall from an anomalous septal branch arising from the right coronary artery. The case illustrates that minor variations in coronary anatomy may have important effects on the prognosis of a patient. PMID- 8221849 TI - Conservative management of guidewire coronary artery perforation with pericardial effusion during angioplasty for acute inferior myocardial infarction. AB - A wire perforation of a totally occluded right coronary artery was treated successfully with prolonged balloon inflation. This non-surgical management is preferred if physiologic and anatomic conditions are suitable. PMID- 8221850 TI - Hemi-balloon dislodgement during a percutaneous balloon pericardial window procedure: removal using a second pericardial catheter. AB - We describe retrieval of a balloon fragment from the pericardial space during a percutaneous balloon pericardial window procedure. A second pericardial catheter was placed, and the transfer wire was snared. The second catheter was then used to push the balloon fragment back through the pericardium and out to the skin. PMID- 8221851 TI - Balloon rupture during peripheral stent implantation: a new technique for balloon retrieval. AB - Balloon rupture is a known technical problem with implantation of vascular stents. In most cases, the ruptured balloon can be retrieved with simple maneuvers. In this case report, a peripheral balloon became trapped within an undeployed peripheral vascular stent and could not be removed by application of standard maneuvers. A novel approach to balloon retrieval was devised and undertaken with success. PMID- 8221852 TI - Pseudodissection of the coronary artery: a variation caused by interventional tools. AB - During PTCA immediate decisions often must be made on the basis of a less than optimum data set. We present a combination of factors which produce an incorrect perception of a coronary artery dissection. This potential must be understood by the interventionalist to avoid misdiagnosis and inappropriate therapeutic maneuvers. PMID- 8221853 TI - Successful prolonged local infusion of urokinase for the hypothenar hammer syndrome. AB - Ulnar artery thrombosis secondary to trauma, also known as the hypothenar hammer syndrome, has been most commonly been treated by surgical techniques. Applying concepts of local lysis of thrombotic occlusion used in cardiac and peripheral vascular intervention, we describe a method of correction of the hypothenar hammer syndrome by prolonged urokinase infusion. PMID- 8221854 TI - Sinus of Valsalva aneurysms involving both coronary ostia. AB - Sinus of Valsalva aneurysms are a rare cause of angina or myocardial infarction. We describe a patient with unstable angina and massive unruptured aneurysms of both coronary sinuses causing severe distortion of both coronary ostia, to our knowledge not previously reported. This unusual patient underwent aortic valve replacement, ascending aortic repair, and coronary artery bypass. At 11-year follow-up, she had developed severe mitral stenosis and prosthetic aortic valve stenosis. Review of the literature reveals 30 reported cases of Sinus of Valsalva aneurysm complicated by coronary insufficiency or infarction, involving either the left (20 cases) or the right coronary sinus (10 cases). We emphasize the poor outcome of these patients, especially if treated conservatively. PMID- 8221855 TI - Pulmonary artery hemodynamic response to proximal balloon dilatation of a large dominant right coronary artery. AB - The human right ventricle has a relatively low tolerance to even short periods of ischemia. During angioplasty of the right coronary artery, pulmonary artery pressure typically rises due to an increase in right ventricular afterload caused by left ventricular dysfunction during ischemia. We have presented two cases of angioplasty in the early proximal portion of a large, highly dominant right coronary artery. Pulmonary artery and systemic pressure fell during balloon inflation probably secondary to acute severe right ventricular failure, though an interaction with left ventricular dysfunction cannot be excluded. These observations do emphasize the hemodynamic and combined ventricular consequences of proximal angioplasty in a large, dominant right coronary artery, particularly in the setting of preexisting left ventricular dysfunction. PMID- 8221856 TI - Sources of error in quantitative coronary angiography. AB - Many studies have reported the accuracy of quantitative coronary angiography (QCA) based on experiments using moderated-size phantoms imaged under unrealistic radiographic conditions. However, these observations may not be generalizable to the setting of clinical angiography. To determine QCA accuracy in a realistic radiographic setting and evaluate the impact of the x-ray system line spread function, plexiglass phantoms were imaged inside and out of a human thorax. A realistic radiographic background was associated with a 38% increase in variability of results (p < 0.05). Low concentrations of contrast and large image intensifier input screens were associated with significantly larger errors and variability in results (p < 0.05). There was a systematic overestimation of diameter in the smallest phantom. A mathematical model of the x-ray line spread function was developed that explains the observed overestimation of the smallest phantom and provide a rational approach for correction of the line spread function for QCA. Many factors encountered in clinical coronary angiography such as nonuniform radiographic background, low concentrations of contrast, and small vessel diameters have a significant adverse impact on the accuracy and/or variability of gradient-based edge detection QCA systems. PMID- 8221857 TI - Catheter lavage and drainage of pneumococcal pericarditis. AB - An 88 year old woman with streptococcal pneumonia developed purulent pericarditis and cardiac tamponade despite treatment with antibiotics. Percutaneous pericardial drainage was effected with a 6 French pigtail catheter inserted via the subxyphoid approach. Catheter drainage was continued for 7 days in conjunction with systemic antibiotics. Catheter patency was maintained with antibiotic lavage. Immediate hemodynamic improvement followed the initial pericardial drainage. Fever, leukocytosis, and sepsis resolved during the course of therapy. The patient recovered fully from the closed space bacterial infection without additional surgical drainage. There has been no recurrence of streptococcal infection and no echocardiographic evidence of recurrent pericardial effusion after 3 months of follow-up. Indwelling catheter drainage combined with antibiotics may be an effective substitute for surgical drainage in the treatment of streptococcal pericarditis. PMID- 8221858 TI - Internal balloon tamponade: a non-surgical method for removal of accidentally placed sheaths from the subclavian artery. AB - One of the possible complications of subclavian vein puncture is accidental puncture of the subclavian artery. If this is not noted immediately after the puncture, insertion of a large bore sheath in the subclavian artery is likely to follow. We describe our experience with a new method that enables successful and safe removal of such sheaths without notable blood loss, in three patients. PMID- 8221859 TI - Quantitative demonstration of dipyridamole-induced coronary steal and alteration by angioplasty in man: analysis by simultaneous, continuous dual Doppler spectral flow velocity. AB - In the course of studying the effects of coronary angioplasty on branch vessel flow using two Doppler flow velocity guidewires, we quantitated simultaneous blood flow responses proximal and distal to a stenosis. The alterations of flow documented a horizontal epicardial steal induced during dipyridamole hyperemia, hyperemic flow reversal by intravenous aminophylline, and subsequent normalization of distal hyperemia after endoluminal enlargement by successful angioplasty. The quantitative physiology of the patient described here confirms one postulated mechanism of abnormal myocardial perfusion stress scintigraphy. Continuous dual flowire spectral coronary flow determinations appear to be a valuable method in verifying postulated mechanisms of various pharmacologic and mechanical stimuli influencing coronary blood flow in patients with atherosclerotic coronary artery disease. PMID- 8221860 TI - Correct positioning for cardiac angiography: insights from MRI. AB - The recommended angiographic projections for optimally demonstrating various cardiac structures vary, and do not always achieve their aim. To determine the most appropriate radiographic projections to optimally demonstrate cardiac structures, we reviewed the orientations of major cardiac structures and the associated great vessels, as demonstrated by axial MRI. Measurements were made from 187 MRI examinations of the heart. These measurements confirmed that the optimum angiographic projections for various cardiac structures are at variance with a number of the recommended views. Recommendations for the angiographic projections most likely to provide optimum display of various cardiac structures and associated great vessels in different patient populations are presented. PMID- 8221861 TI - Experimental validation of geometric and densitometric coronary measurements on the new generation Cardiovascular Angiography Analysis System (CAAS II). AB - Computer-assisted contour detection and videodensitometric cross sectional area assessment of coronary artery obstructions on the CAAS II system were validated in vitro and in vivo by angiographic cinefilm recording and automated measurement of stenosis phantoms (luminal diameter 0.5, 0.7, 1.0, 1.4, 1.9 mm) which were first inserted in a plexiglass model and then serially implanted in swine coronary arteries. "Obstruction diameter" (OD) and "obstruction area" (OA) values obtained from 10 in vitro and 19 in vivo images at the site of the artificial stenoses were compared with the true phantom dimensions. The in vitro assessment of OD yielded an accuracy of 0.00 +/- 0.11 mm (correlation coefficient: r = 0.98, y = 0.18 + 0.82x, standard error of estimate: SEE = 0.08), whereas the in vivo measurement of OD gave an accuracy of -0.01 +/- 0.18 mm (r = 0.94, y = 0.22 + 0.82x, SEE = 0.15). The assessment of OA gave an accuracy of -0.08 +/- 0.21 mm2 in vitro (r = 0.97, y = 0.08 + 0.99x, SEE = 0.22) and -0.22 +/- 0.32 mm2 in vivo (r = 0.95, y = 0.21 + 1.01x, SEE = 0.33). The mean reproducibility was +/- 0.09 mm for geometric measurements and +/- 0.21 mm2 for videodensitometric assessments, respectively. Thus, due to inherent limitations of the imaging chain, the reliability of geometric coronary measurements is still far superior to videodensitometric assessments of vessel cross sectional areas. PMID- 8221862 TI - High-speed rotational ablation (Rotablator) for unfavorable lesions in peripheral arteries. AB - A strategy of using a single, conservatively sized percutaneous transluminal rotational ablation device with or without adjunctive balloon angioplasty was employed in 18 vessels in 17 patients selected because of unfavorable lesion morphology for balloon angioplasty alone. Fifteen patients had lower extremity occlusions and/or heavily calcified lesions and two patients had ostial renal artery stenoses. We were able to achieve a 94% (17/18 lesions) technical success rate, and a 94% (16/17 patients) clinical success rate. In the patients with lower extremity lesions, the baseline ankle-brachial blood pressure index increased from 0.55 +/- 0.15 to 0.90 +/- 0.19 (p < .001) 1 day after the procedure. Follow-up at 6.8 +/- 2.8 months revealed clinical evidence of restenosis in only one patient. We conclude that a cost-effective strategy of treating unfavorable lesions with a single Rotablator burr and adjunctive balloon angioplasty is safe and effective. PMID- 8221863 TI - Percutaneous rotational thrombectomy reduces acute reocclusion in an animal model of acute coronary thrombosis. AB - This study was designed to compare acute reocclusion rates after treatment of acute coronary thrombosis with a percutaneous thrombectomy device or standard balloon angioplasty. Our group has previously reported on the rationale and development of a mechanical device for the treatment of intra-arterial thrombosis. This device removes fibrin from thrombus, allowing for dissolution of the cellular elements of the thrombus. Theoretically, thrombus removal (as opposed to displacement) might result in a lower rate of acute rethrombosis. The present study utilizes the device percutaneously in the coronary arteries of closed chest swine and compares recanalization and reocclusion rates with standard balloon angioplasty. Twenty-six animals with total thrombotic coronary occlusions were treated; 13 with each device. Reocclusion rates with the thrombectomy device were significantly reduced at 60 min and 120 min after recanalization (p < 0.02), and the mean time to reocclusion was prolonged by 45 min (p = 0.07). Technical problems included poor handling characteristics in early prototypes and stress fractures secondary to improper use. Changes in catheter design and operator protocols have largely eliminated these problems. We conclude that this study demonstrates the feasibility of percutaneous mechanical thrombectomy in the coronary arteries and that reocclusion rates after recanalization of thrombotic occlusions compare favorably to standard angioplasty. PMID- 8221864 TI - Transluminal extraction atherectomy and adjunctive balloon angioplasty for restenosis after Palmaz-Schatz coronary stent implantation. AB - Two patients who developed restenosis after implantation of Palmaz-Schatz coronary stents were successfully treated by transluminal extraction atherectomy and there has been no recurrence on follow-up angiograms. The optimum strategy for managing restenosis after coronary stenting remains unclear, but transluminal extraction atherectomy appears to be a safe and effective option. PMID- 8221865 TI - Treatment of simple and complex coronary stenosis using rotational ablation followed by low pressure balloon angioplasty. AB - High speed coronary rotational ablation followed by adjunctive low pressure coronary balloon angioplasty was used in 166 patients. Complex lesions were present in 63% of patients. Clinical success was achieved in 157 patients (95%). Angiographic success rate was 10% in A lesions, 95% in B1 lesions, 98% in B2 lesions, and 93% in C lesions. Mean vessel lumen diameter has increased from 0.4 mm to 1.4 mm after rotational ablation and to 2.3 mm following adjunctive balloon angioplasty. Left ventricular dysfunction, the presence of total occlusion, and the severity of stenosis before the procedure were predictive of clinical failure. There were three abrupt vessel closures (1.8%) with one Q wave myocardial infarction. Non-Q wave myocardial infarction occurred in 14 patients (8.4%) and three patients died during hospitalisation (1.8%). We conclude that rotational atherectomy followed by adjunctive low pressure balloon angioplasty gives comparable high success rates in different coronary lesions. PMID- 8221866 TI - Percutaneous balloon valvuloplasty in mirror-image dextrocardia and rheumatic mitral stenosis. AB - A 28-year-old woman with mirror-image dextrocardia and severe rheumatic mitral stenosis underwent successful percutaneous balloon mitral valvuloplasty (PBMV). Standard technique was altered to include transseptal catheterization via the left femoral vein and inter-atrial septal puncture with the transseptal needle rotated to a 7 o'clock position. The predilatation transmitral gradient (mean) of 16 mmHg decreased to 5 mmHg following PBMV. The valve area improved from 1.0 cm2 to 2.6 cm2. There were no complications. This case illustrates that transseptal catheterization can be accomplished safely in patients with unusual cardiac anatomy and mitral valvuloplasty can be performed in patients with mirror-image dextrocardia and rheumatic mitral stenosis. PMID- 8221867 TI - Prolonged intracoronary urokinase infusion for acute stent thrombosis. PMID- 8221868 TI - Ventricular free wall and septal rupture (double rupture): a "pseudocomplication" during post-infarction laser angioplasty. AB - We report the rare occurrence of double rupture of the myocardium occurring immediately following successful laser recanalization of an occluded right coronary artery in a 72-year-old woman 5 days following infero-posterior myocardial infarction. PMID- 8221869 TI - Prolonged heparin therapy for occlusive intracoronary thrombus. AB - The presence of intracoronary thrombus is associated with increased complications during coronary angioplasty. Such thrombus may also mimic the appearance of a critical stenosis. We report a case of nearly occlusive intracoronary thrombus which resolved after prolonged heparin therapy, revealing only a minimal underlying stenosis. The recognition and treatment of this entity is discussed. PMID- 8221870 TI - Giant left main pseudoaneurysm obstructing the left anterior descending coronary artery demonstrated by transesophageal echocardiography and coronary angiography. AB - Left main pseudoaneurysm is a rare angiographic finding in young adults. We describe a 23-year-old male who presented with an acute anterior wall myocardial infarction. Coronary angiography and transesophageal echocardiography reveal a huge aneurysm of the distal left main obstructing the left anterior descending artery (LAD), which was defined by history as a pseudoaneurysm. The pseudoaneurysm was successfully resected and mammary artery was used to repair the distal left main and to bypass the LAD. PMID- 8221871 TI - Closure of a coronary cameral fistula following endomyocardial biopsies in a cardiac transplant patient with a detachable balloon. AB - Endomyocardial biopsy is the reference standard for the diagnosis of cardiac allograft rejection and is performed frequently in cardiac transplant patients. Biopsies are taken percutaneously via the right internal jugular or femoral vein. Fistulas from coronary arteries into the right ventricle following endomyocardial biopsies are a relatively frequent finding. Most of these fistulas are small angiographically, haemodynamically insignificant, and tend to disappear. We report a case of a haemodynamic significant coronary fistula, closed by a percutaneously introduced detachable balloon. PMID- 8221872 TI - Transcoronary cardiac pacing during myocardial ischemia. AB - Severe bradyarrhythmias are a rare but potentially life threatening complication of percutaneous transluminal coronary angioplasty (PTCA). Previous work has outlined a technique for coronary pacing using the angioplasty guidewire. To examine the effectiveness of this technique during severe ischemia, seven swine underwent placement of an unmodified 0.014 inch angioplasty guidewire and 3.0 mm balloon catheter in the left anterior descending (LAD) artery. Baseline pacing thresholds were obtained. Pacing was begun at twice diastolic threshold and ischemia was produced by balloon inflation. Repeat capture thresholds were obtained after 1 and 8 minutes of ischemia. Transcoronary pacing was successfully performed in all seven animals and was continued for a mean of 13.8 +/- 1.5 minutes. The baseline capture threshold was 4.0 +/- 0.5 mA. The mean capture threshold was 3.3 +/- 0.3 mA and 4.5 +/- 0.9 mA at 1 and 8 minutes of ischemia, respectively. We conclude that transcoronary pacing using the angioplasty guidewire can be successfully performed during myocardial ischemia and may serve as a reliable backup system during interventional procedures complicated by bradyarrhythmias. PMID- 8221873 TI - Retrieval of a migrated coronary stent by means of an alligator forceps catheter. AB - Stent migration and embolization are well-known complications of intracoronary stenting with balloon-mounted stents. During an elective stenting procedure of a proximal right coronary artery stenosis, a 3.5 mm Wiktor stent (Medtronic Inc., Minneapolis) was displaced from its delivery balloon. The guiding catheter and the delivery system were withdrawn, leaving the stent around a 3 m 0.014 inch High Torque floppy guidewire (ACS, Santa Clara, CA) in the abdominal aorta. An 40 cm 5F Alligator Forceps catheter (Cook OB/Gyn., Spencer, IN), introduced through a cut-off 8F coronary guiding catheter, allowed improved torque control of the retrieval catheter and a safe and successful withdrawal of the stent through the arterial introducer sheet. PMID- 8221874 TI - Use of an intracoronary stent to control intrapericardial bleeding during coronary artery rupture complicating coronary angioplasty. AB - Coronary artery rupture is a rare complication of percutaneous transvenous coronary angioplasty (PTCA) usually requiring urgent cardiac surgery and often resulting in a poor outcome including death. We report, for the first time, the use of an intracoronary stent to control intrapericardial bleeding following the development of a coronary artery rupture during PTCA. Cardiac surgery was avoided and the patient was discharged well 1 week after the procedure. PMID- 8221875 TI - Percutaneous transradial artery approach for coronary stent implantation. AB - A new approach for implantation of Palmaz Schatz coronary stents is reported. We describe the technique and rationale of coronary stenting with miniaturized angioplasty equipment via the radial artery. This technique is illustrated in three patients. One patient underwent Palmaz Schatz stent implantation for a saphenous vene coronary bypass graft stenosis, the second patient for a restenosis in the anterior descending coronary artery after atherectomy, and the third patient for a second restenosis after balloon angioplasty in the circumflex coronary artery. PMID- 8221876 TI - What ultrafast CT (UFCT) can and cannot do in coronary atherosclerosis. PMID- 8221877 TI - Percutaneous balloon mitral valvotomy in patients with left atrial thrombus. PMID- 8221878 TI - TEE-guided percutaneous balloon mitral valvuloplasty. PMID- 8221879 TI - Use of the nitinol .014 coronary guidewire during directional coronary atherectomy. PMID- 8221880 TI - Placement of "biliary" stents in saphenous vein coronary bypass grafts. AB - Angioplasty of coronary saphenous vein grafts has been associated with less favorable results than in native coronary arteries owing to a higher acute complication rate and an increased incidence of restenosis. We placed 16 nonarticulated balloon expandable stainless steel "biliary" stents at the sites of 13 stenotic or occluded aortocoronary saphenous vein graft lesions in 11 patients. All of the lesions were stented successfully. There were no instances of stent thrombosis or stent embolism. The percent diameter stenosis was reduced from 85.5 +/- 14.1% to 3.5 +/- 4.4% (p < .001), and the minimal lumen diameter of the lesion increased from 0.7 +/- 0.7 mm at baseline to 3.7 +/- 0.2 mm (p < .001) after stent placement. We conclude that this noncoronary stent appears to be safe and effective for treating saphenous vein coronary bypass grafts. The high procedural success rate and excellent angiographic results are very encouraging, while the restenosis rates remain to be determined. PMID- 8221881 TI - Clinically guided closure of femoral arterial pseudoaneurysms complicating cardiac catheterization and coronary angioplasty. AB - Femoral artery pseudoaneurysm formation is a significant problem in patients undergoing cardiac catheterization and interventional cardiac procedures. It is especially more common with the use of anticoagulant and antiplatelet therapy and the use of intracoronary stents. We describe our initial experience with clinically guided bedside compression of femoral pseudoaneurysms in such patients. Eleven patients, 10 undergoing coronary angioplasty (including 3 with intracoronary stents) and 1 undergoing diagnostic cardiac catheterization, developed a femoral pseudoaneurysm. All patients had a femoral bruit and 9 had an expansile groin hematoma. The diagnosis was confirmed in each case by Doppler ultrasound. Seven patients were receiving heparin while 4 were on oral anticoagulants at the time of detection of the pseudoaneurysm. These patients underwent clinically guided graded external compression to close the pseudoaneurysm neck while maintaining femoral arterial flow. External compression for 104.1 +/- 63 min resulted in successful clinical resolution of pseudoaneurysm in all patients without complications. The results were confirmed by Doppler ultrasound at least 12 hr later. Bedside compression of femoral pseudoaneurysms guided by clinical clues is simple and appears to be an effective and safe technique to manage this iatrogenic problem. PMID- 8221882 TI - Novel intron-encoded small nucleolar RNAs. PMID- 8221883 TI - scat+ is a selfish gene analogous to Medea of Tribolium castaneum. PMID- 8221884 TI - A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion. AB - The SNARE hypothesis holds that a transport vesicle chooses its target for fusion when a soluble NSF attachment protein (SNAP) receptor on the vesicle (v-SNARE) pairs with its cognate t-SNARE at the target membrane. Three synaptosomal membrane proteins have previously been identified: syntaxin, SNAP-25 (t-SNAREs), and vesicle-associated membrane protein (VAMP) (v-SNARE); all assemble with SNAPs and NSF into 20S fusion particles. We now report that in the absence of SNAP and NSF, these three SNAREs form a stable complex that can also bind synaptotagmin. Synaptotagmin is displaced by alpha-SNAP, suggesting that these two proteins share binding sites on the SNARE complex and implying that synaptotagmin operates as a "clamp" to prevent fusion from proceeding in the absence of a signal. The alpha-SNAP-SNARE complex can bind NSF, and NSF-dependent hydrolysis of ATP dissociates the complex, separating syntaxin, SNAP-25, and VAMP. ATP hydrolysis by NSF may provide motion to initiate bilayer fusion. PMID- 8221885 TI - Identification of katanin, an ATPase that severs and disassembles stable microtubules. AB - Eukaryotic cells rapidly reorganize their microtubule cytoskeleton during the cell cycle, differentiation, and cell migration. In this study, we have purified a heterodimeric protein, katanin, that severs and disassembles microtubules to tubulin dimers. The disassembled tubulin can repolymerize, indicating that it is not irreversibly modified or denatured in the reaction. Katanin is a microtubule stimulated ATPase and requires ATP hydrolysis to sever microtubules. Katanin represents a novel type of enzyme that utilizes energy from nucleotide hydrolysis to break tubulin-tubulin bonds within a microtubule polymer, a process that may aid in disassembling complex microtubule arrays within cells. PMID- 8221886 TI - Mammalian achaete-scute homolog 1 is required for the early development of olfactory and autonomic neurons. AB - The mouse Mash-1 gene, like its Drosophila homologs of the achaete-scute complex (AS-C), encodes a transcription factor expressed in neural precursors. We created a null allele of this gene by homologous recombination in embryonic stem cells. Mice homozygous for the mutation die at birth with apparent breathing and feeding defects. The brain and spinal cord of the mutants appear normal, but their olfactory epithelium and sympathetic, parasympathetic, and enteric ganglia are severely affected. In the olfactory epithelium, neuronal progenitors die at an early stage, whereas the nonneuronal supporting cells are present. In sympathetic ganglia, the mutation arrests the development of neuronal precursors, preventing the generation of sympathetic neurons, but does not affect glial precursor cells. These observations suggest that Mash-1, like its Drosophila homologs of the AS-C, controls a basic operation in development of neuronal progenitors in distinct neural lineages. PMID- 8221887 TI - Stepwise dismantling of adenovirus 2 during entry into cells. AB - Adenoviruses enter their host cells by receptor-mediated endocytosis and acid activated penetration from endosomes into the cytosol and deliver their DNA genome into the nucleus. Our results show that incoming adenovirus type 2 particles undergo a stepwise disassembly program necessary to allow progress of the virus in the entry pathway and release of the genome into the nucleus. The fibers are released, the penton base structures dissociated, the proteins connecting the DNA to the inside surface of the capsid degraded or shed, and the capsid-stabilizing minor proteins eliminated. The uncoating process starts immediately upon endocytic uptake with the loss of fibers and ends with the uptake of dissociated hexon proteins and DNA into the nucleus. PMID- 8221888 TI - MKP-1 (3CH134), an immediate early gene product, is a dual specificity phosphatase that dephosphorylates MAP kinase in vivo. AB - Mitogenic stimulation of cells induces rapid and transient activation of MAP kinases. Here we report that a growth factor-inducible gene, 3CH134, encodes a dual specificity phosphatase that dephosphorylates and inactivates p42MAPK both in vitro and in vivo. In vitro, 3CH134 protein dephosphorylates both T183 and Y185 in p42MAPK. In serum-stimulated normal fibroblasts, the kinetics of inactivation of p42MAPK coincides with the appearance of newly synthesized 3CH134 protein, and the protein synthesis inhibitor cycloheximide leads to persistent activation of MAP kinase. Expression of 3CH134 in COS cells leads to selective dephosphorylation of p42MAPK from the spectrum of phosphotyrosyl proteins. 3CH134 blocks phosphorylation and activation of p42MAPK mediated by serum, oncogenic Ras, or activated Raf, whereas the catalytically inactive mutant of the phosphatase, Cys-258-->Ser, augments MAP kinase phosphorylation under similar conditions. The mutant 3CH134 protein also forms a physical complex with the phosphorylated form of p42MAPK. These findings suggest that 3CH134 is a physiological MAP kinase phosphatase; we propose the name MKP-1 for this phosphatase. PMID- 8221889 TI - The HPV-16 E6 and E6-AP complex functions as a ubiquitin-protein ligase in the ubiquitination of p53. AB - The ubiquitin-dependent proteolytic pathway plays a major role in selective protein degradation. Ubiquitination of proteins requires the sequential action of the ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzymes (E2), and in some cases ubiquitin-protein ligases (E3s). The oncogenic human papillomavirus (HPV) types 16 and 18 utilize this cellular proteolytic system to target the tumor suppressor protein p53. The HPV E6 oncoprotein binds to a cellular protein of 100 kd, termed E6-associated protein (E6-AP). The E6-E6-AP complex specifically interacts with p53, resulting in the rapid ubiquitin-dependent degradation of p53. Here we report the purification and identification of the factors necessary for the E6-E6-AP-mediated ubiquitination of p53. The ubiquitination of p53 requires the E1 enzyme and a novel E2 in mammalian cells, while E3 activity is conferred by the E6-E6-AP complex. Furthermore, E6-AP appears to have ubiquitin-protein ligase activity in the absence of E6. PMID- 8221890 TI - High level expression of the Activator transposase gene inhibits the excision of Dissociation in tobacco cotyledons. AB - A fusion of the strong cauliflower mosaic virus 35S promoter to the Activator (Ac) transposase (TPASE) gene does not trigger excision of Dissociation (Ds) continuously during tobacco cotyledon development, although once activated, the 35S promoter remains active throughout embryogeny. Epistasis studies where 35S:TPASE is in trans with later-acting fusions indicate that transient effectiveness for excision results from this fusion inhibiting its own action and that of other TPASE sources. Inhibition depends on the strength of TPASE expression, since fusions of the 35S promoter to a TPASE cDNA accumulate 30-fold lower amounts of TPASE mRNA than the 35S:TPASE gene fusion and do not inhibit excision. We discuss the role of TPASE levels in the curious relationship between Ac activity and Ac dosage in maize. PMID- 8221891 TI - Drosophila TAFII40 interacts with both a VP16 activation domain and the basal transcription factor TFIIB. AB - Enhancement of RNA polymerase II transcription by the viral transactivator VP16 requires the TFIID complex, which consists of the TATA-binding protein (TBP) and TBP-associated factors (TAFs). Here we report the molecular cloning, expression, and biochemical characterization of Drosophila TAFII40 (dTAFII40), a subunit of TFIID. In vitro protein-protein interaction assays revealed direct binding between dTAFII40 and a 39 amino acid VP16 activation domain. In addition, affinity chromatography indicated a direct binding of the basal factor TFIIB to immobilized dTAFII40. Since VP16 also binds TFIIB, our results suggest a ternary interaction among an activator, a coactivator, and a basal transcription factor. Antibodies directed against dTAFII40 inhibited activation by GAL4-VP16 without affecting basal transcription. These results, taken together with previous studies of Sp1 and dTAFII110, establish that different activators interact with distinct TAFs in the TFIID complex and that TAFs can contact both activators and basal factors. PMID- 8221892 TI - Targeting of SIR1 protein establishes transcriptional silencing at HM loci and telomeres in yeast. AB - Previous studies suggest that the yeast SIR1 protein is involved in the establishment of transcriptional silencing at the HM mating-type loci. Here we show that a GAL4 DNA-binding domain-SIR1 hybrid protein (GBD-SIR1), when targeted to an HMR locus containing GAL4-binding sites (UASG), can establish silencing and bypass the requirement for the silencer element HMR-E. Silencing mediated by GBD SIR1 requires the trans-acting factors that normally participate in repression, namely, SIR2, SIR3, SIR4, and histone H4. However, GBD hybrids with SIR2, SIR3, or SIR4 cannot establish silencing. Telomeric silencing, which does not require SIR1 and is normally unstable, is greatly improved by tethering GBD-SIR1 to the telomere. These experiments support a model in which native SIR1 protein is brought to the HM loci by proteins bound to the silencers. Telomeres appear to lack the ability to recruit SIR1, and that is why telomeric silencing is unstable. PMID- 8221893 TI - SIR3 and SIR4 proteins are required for the positioning and integrity of yeast telomeres. AB - Heritable inactivation of genes occurs in specific chromosomal domains located at the silent mating type loci and at telomeres of S. cerevisiae. The SIR genes (for silent information regulators) are trans-acting factors required for this repression mechanism. We show here that the SIR3 and SIR4 gene products have a sub-nuclear localization similar to the telomere-associated RAP1 protein, which is found primarily in foci at the nuclear periphery of fixed yeast spheroplasts. In strains deficient for either SIR3 or SIR4, telomeres lose their perinuclear localization, as monitored by RAP1 immunofluorescence. The length of the telomeric repeat shortens in sir3 and sir4 mutant strains, and the mitotic stability of chromosome V is reduced. These data suggest that SIR3 and SIR4 are required for both the integrity and subnuclear localization of yeast telomeres, the loss of which correlates with loss of telomere-associated gene repression. PMID- 8221894 TI - Protein-protein interactions in gene regulation: the cAMP-CRP complex sets the specificity of a second DNA-binding protein, the CytR repressor. AB - Maximal repression by the CytR protein depends on the formation of nucleoprotein complexes in which CytR interacts with DNA and with cAMP-cAMP receptor protein (CRP). Here we demonstrate that CytR regulates transcription from deoP2 promoters in which the entire CytR recognition sequence has been eliminated. Furthermore, CytR proteins deleted for the DNA-binding domain repress deoP2 in vivo and interact with deoP2 in vitro in a strictly cAMP-CRP-dependent fashion. These experiments show that the site of action of CytR can be specified by protein protein interactions to cAMP-CRP, whereas CytR-DNA interactions may primarily serve to stabilize the nucleo-protein complex. This type of specificity mechanism may represent a general concept in the recruitment of DNA-binding proteins in combinatorial regulatory systems. PMID- 8221895 TI - The crystal structure of the estrogen receptor DNA-binding domain bound to DNA: how receptors discriminate between their response elements. AB - The nuclear hormone receptors are a superfamily of ligand-activated DNA-binding transcription factors. We have determined the crystal structure (at 2.4 A) of the fully specific complex between the DNA-binding domain from the estrogen receptor and DNA. The protein binds as a symmetrical dimer to its palindromic binding site consisting of two 6 bp consensus half sites with three intervening base pairs. This structure reveals how the protein recognizes its own half site sequence rather than that of the related glucocorticoid receptor, which differs by only two base pairs. Since all nuclear hormone receptors recognize one or the other of these two consensus half site sequences, this recognition mechanism applies generally to the whole receptor family. PMID- 8221896 TI - FGF-4 replaces the apical ectodermal ridge and directs outgrowth and patterning of the limb. AB - The apical ectodermal ridge plays a key role in limb development. We show that recombinant FGF-4 can substitute for the ridge to provide all the signals necessary for virtually complete outgrowth and patterning of the chick limb. FGF 4 stimulates proliferation of cells in the distal mesenchyme and maintains a signal from the posterior to the distal mesenchyme that appears to be required for elaboration of skeletal elements in the normal proximodistal sequence. Moreover, retinoic acid, which is capable of providing polarizing activity, can supply this signal. This suggests that polarizing activity plays a role in patterning along the proximodistal axis, in addition to its well-established role in anteroposterior patterning. Taken together, the data suggest a simple mechanism whereby FGF-4 links growth and pattern formation during limb development. PMID- 8221897 TI - Use of microbial spores as a biocatalyst. AB - Endospores of a bacterium Bacillus subtilis and ascospores of a yeast Saccharomyces cerevisiae contained almost all the activities for the same enzymes as vegetative cells. The biotechnological potential of spores was studied by selecting adenosine 5'-triphosphatase and alkaline phosphatase in bacterial and yeast spores, respectively, as model enzymes. The activity of both enzymes was efficiently expressed when the spores were treated by physical (sonication or electric field pulse) and chemical (organic solvents or detergents) methods. The yeast spores were immobilized in polyacrylamide gel without any appreciable loss of activity. The immobilized spores were packed in a column and used successfully for the continuous reactions of alkaline phosphatase and glyoxalase I. The microbial spores were confirmed to be promising as a biocatalyst for the production of useful chemicals in bioreactor systems. PMID- 8221898 TI - Problems in scale-up of biotechnology production processes. AB - The unique nature of biotechnology processes adds to the complexity and difficulty of scale-up. Successful scale-up means a shortened cycle to full-scale production, competitive advantage, and cost savings. The many pitfalls as well as actual and potential scale-up problems are reviewed. Emphasis is placed on covering all areas of concern in planning, executing, and documenting key studies. Needs in technology transfer are discussed and regulatory requirements are incorporated into scale-up needs. A review of the recent literature is coupled with actual case studies; problem avoidance is stressed. Problems in asepsis, in construction, and in validation are discussed and potential solutions given. Organizational problems are noted. Finally, checklists are given for project planning, for a safety audit, and for timely attainment of successful scale-up. Eighty-two references are included. PMID- 8221899 TI - Ontogenesis of F-actin in hair cells. AB - This report describes the ontogenesis of cochlear stereocilia using scanning electron microscopy for analysis of cilia appearance, and fluorescence microscopy of phalloidin, a label for F-actin, to determine the maturation of the cilia framework. Surface and frozen-sectioned preparations of the otic capsule were obtained from several stages of rat pup development beginning at the 16th gestational day and at various stages until adulthood. In the earliest stage investigated, strong fluorescence labeling was visible on the apical part of Kolliker's organ, revealing a reticular outline of cell junctions. Hair cells started to differentiate at the 18th day of gestation from cells within the primordial receptor area. Phalloidin labeling revealed a sequential appearance of F-actin as the hair cells differentiated from the cells with the Kolliker's organ. The differentiation of receptor cells occurred first with the appearance of a junctional complex between the hair cell and the surrounding cells. Then a cuticular plate appeared followed by the progressive emergence of stereocilia. The F-actin labeling also revealed a progressive differentiation of receptor cells from the cochlear base to its apex. There was also an inner to outer hair cell developmental gradient of label. Inner hair cells developed stereocilia before outer hair cells. The third row of outer hair cells was the last to acquire stereocilia. The adult pattern of stereocilia was reached around the 6th postnatal day. We conclude that the appearance of actin filaments in developing receptor cells and the emergence of stereocilia can be regraded as markers for correlating function and other structural differentiation. PMID- 8221900 TI - Fimbrin localized to an insoluble cytoskeletal fraction is constitutively phosphorylated on its headpiece domain in adherent macrophages. AB - The actin-bundling protein fimbrin is homologous to 1-plastin, a 65kD phosphoprotein expressed in leukocytes and transformed cells [de Arruda et al., J. Cell Biol. 111, 1069-1080]. Because fimbrin is present in cell adhesion sites, we studied the phosphorylation state of fimbrin and its distribution in macrophages sequentially extracted with Triton-X-100 (soluble fraction), Tween 40 deoxy-cholate (cytoskeletal fraction), and SDS (insoluble cytoskeletal fraction). The approximate distribution of fimbrin and actin among these fractions was found to be: 65% fimbrin/55% actin in the soluble fraction, 30% fimbrin/20% actin in the cytoskeletal fraction, and 5% fimbrin/25% actin in the insoluble cytoskeletal fraction. PMA did not alter this distribution. Fluorescence microscopy of acetone extracted macrophages showed that actin is concentrated in podosomes at the substratum interface and is diffusely distributed throughout the remainder of the cell. Fimbrin colocalizes with actin in podosomes and also exhibits a punctate distribution in the cytoplasm that overlaps with actin. In Tween 40/DOC-extracted cells, podosomes remain, and fimbrin also exhibits a punctate distribution along actin filaments. Metabolic 32PO4 labeling revealed that fimbrin is constitutively phosphorylated and that phosphorylated fimbrin is concentrated in the insoluble cytoskeletal fraction. PMA increased the relative levels of fimbrin phosphorylation twofold but did not alter the pattern of fimbrin fluorescence or the distribution of phosphorylated fimbrin. Limited trypsin digestion and phosphoamino acid analysis demonstrated that phosphorylation occurs specifically on serine residues within the 10kD headpiece domain of fimbrin. Phosphorylation of the headpiece domain could regulate the actin binding and bundling properties of fimbrin, or it could regulate the interaction of fimbrin with other proteins. PMID- 8221901 TI - Sulphonate buffers affect the recovery of microtubule-associated proteins MAP1 and MAP2: evidence that MAP1A promotes microtubule assembly. AB - The influence of two commonly used sulphonate buffers, PIPES and MES, on the in vitro assembly of bovine brain microtubule protein was examined. Microtubule assembly was monitored by turbimetry and, after centrifugation, the polymerised protein was analysed by SDS-PAGE and western blotting. Assembly in MES when compared with PIPES resulted in a higher recovery of microtubule proteins at both pH 6.4 and pH 6.9 and in an altered protein composition. The buffer pH affected the total amount of protein polymerised but did not significantly affect the protein composition. At both pH conditions the recovery of HMW-MAPs was markedly increased in MES buffer and this increase was mostly due to an increase in the amount of MAP1. PMID- 8221902 TI - Perspectives on tubulin isotype function and evolution based on the observation that Tetrahymena thermophila microtubules contain a single alpha- and beta tubulin. AB - We have cloned and sequenced the two beta-tubulin genes of the ciliated protozoan Tetrahymena thermophila. The two genes encode identical 443 amino acid peptides which are 99.7% identical to the beta-tubulin proteins of T. pyriformis and 95% identical to human beta 1 tubulin. T. thermophila contains only one alpha-tubulin gene (Callahan et al., 1984: Cell 36:441-445). Thus, all of the extremely diverse microtubule structures in this unicellular organism can be formed from a single alpha- and a single beta-tubulin peptide. We have also carried out a phylogenetic analysis of 84 complete beta-tubulin peptide sequences. This analysis supports two hypotheses regarding beta-tubulin evolution and function: 1) Multifunctional beta-tubulins are under greater evolutionary constraint than beta-tubulins present in specialized cells or in cells with very few microtubule related functions, which can evolve rapidly; and 2) Cells which form axonemes maintain a homogeneous population of tubulins. PMID- 8221903 TI - Motility of intracellular particles in rat fibroblasts is greatly enhanced by phorbol ester and by over-expression of normal p21N-ras. AB - Particle motility in cultured rat fibroblasts was studied using video-enhanced differential interference contrast microscopy. The average velocity of large bright particles (apparent diameter about 0.5-0.7 micron) was measured in control cells and in cells treated with agents which affected targets related to signal transduction pathways. A Rat-2-derived fibroblast line transfected with a construct containing multiple copies of the N-ras proto-oncogene under the control of dexamethasone-sensitive promoter was used as a main experimental model. Dexamethasone treatment was shown to induce high levels of N-ras expression in these cells. This treatment greatly increased the average particle velocity. At the same time dexamethasone did not influence the particle motility in the non-transfected parent cells and in the cells transfected with a construct which did not contain N-ras. Phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C (PKC), also induced an approximate eightfold increase in the particle rate after several hours of incubation, while sphingosine, an inhibitor of PKC, prevented this activation. Sphingosine alone reduced the particle motility after a 20 min incubation. The particle movements were inhibited also by colcemid. These data show that the activation of N-ras and PKC produced dramatic activation of microtubule-dependent particle motility. A possible role of this activation in signal-induced alterations of cell morphology is discussed. PMID- 8221904 TI - Multiple sites for subtilisin cleavage of tubulin: effects of divalent cations. AB - Limited digestion of pig brain GDP-tubulin by subtilisin was carried out in the presence of Mg2+, Mn2+, Ca2+, Zn2+, or Be2+. Isoelectric focusing, followed by SDS-PAGE, revealed characteristic divalent cation-dependent changes in the alpha- and beta-tubulin cleavage patterns. Previous studies revealed that the beta cleavage pattern is different for heterodimers and microtubules [Lobert and Correia, 1992: Arch. Biochem. Biophys. 296: 152-160]. Divalent cation effects on subtilisin digestion of tubulin indicate different classes of divalent cation binding sites. Western blot analysis locates the proteolytic zone at residue 430 or higher in both subunits for all conditions. Turbidity and electron microscopy reveal that GDP-tubulin cleaved by subtilisin in the presence of Mg2+, Ca2+, or Mn2+ forms sheets of rings. Mn2+ induces ring formation in uncleaved GDP-tubulin. Isotype-depleted tubulin was generated by the removal of class III beta-tubulin using immunoaffinity chromatography. Subtilisin digestion of the depleted fraction and the purified class III beta-tubulin demonstrates that cleavage occurs at three to four distinct sites. Thus, subtilisin-digested tubulin is more heterogeneous than was previously reported and the cleavage sites depend on solution conditions, divalent cations, and the state of assembly. This has important implications for experiments that utilize subtilisin-digested tubulin for studying microtubule-associated protein binding. PMID- 8221905 TI - Site-directed antibodies to tubulin. PMID- 8221906 TI - Near-UV radiation disrupts filamentous actin in lens epithelial cells. AB - Ultraviolet radiation in the near range (UVA) causes lens opacification and disrupts the actin cytoskeleton in rabbit and gray squirrel lenses. Changes were noted using transmission electron microscopy of tangential sections and rhodaminephalloidin fluorescence microscopy of epithelial whole mounts of irradiated and unirradiated lenses, and corresponded with gross cataract formation. Irradiated lenses lacked microfilament polygonal arrays at the inner surface of the apical plasma membrane (i.e., in the cell pole next to the lens fibers) in lens epithelia of both species; a condensed actin bundle was present instead. This bundle, and scattered small actin clumps in the cytoplasm, were identified by immunogold TEM, using a specific antibody and a secondary antibody conjugated with colloidal gold. Similar techniques showed breakdown of tubulin and vimentin, but after longer intervals than for the breakdown of actin. Generalized cytologic damage was also present in epithelial cells, but not in the underlying cortical lens fibers. Damage began to occur after 4 hr of irradiation and became more severe with increased exposure. Shielded controls remained clear, had normal cytology and polygonal arrays, and no clumping of actin filaments. PMID- 8221907 TI - PKC mediates 12(S)-HETE-induced cytoskeletal rearrangement in B16a melanoma cells. AB - The fatty acid 12(S)-HETE may be a new second messenger capable of activating PKC. In tumor cells 12(S)-HETE stimulates cytoskeleton-dependent cellular responses such as adhesion and spreading. Analysis of 12(S)-HETE effects on B16a melanoma cell cytoskeleton revealed reversible rearrangement of microtubules, microfilaments, the actin-binding proteins, vinculin, myosin heavy (MHC) and light chains (MLC), as well as bundling of vimentin intermediate filaments. The alterations in microfilaments and intermediate filaments occurred very rapidly, i.e., 5 min after exposure of tumor cells to 12(S)-HETE. The 12(S)-HETE-induced cytoskeletal alterations were accompanied by centrifugal organelle-translocation. Interestingly, MLC exhibited clear association with the cytoplasmic organelles. Biochemical analysis of the 12(S)-HETE effect indicated a PKC-mediated reversible hyperphosphorylation of MLC, vimentin, and a 130 kD cytoskeletal-associated protein. Optimal effects were obtained after 5 min treatment with 12(S)-HETE at 0.1 microM concentration. 12(S)-HETE pretreatment induced tumor cell spreading on a fibronectin matrix which required the intactness of all three major cytoskeletal components. The spreading process was dependent upon the activity of PKC. Our data suggest that 12(S)-HETE is a physiological stimulant of PKC. Further, it induces rearrangement of the cytoskeleton of tumor cells in interphase resulting in the stimulation of cytoskeleton-dependent cell activity such as spreading. PMID- 8221908 TI - Inhibition of microtubule sliding by Ni2+ and Cd2+: evidence for a differential response of certain microtubule pairs within the bovine sperm axoneme. AB - Bovine sperm, extracted with 0.1% Triton X-100, frozen at -20 degrees C for 48 120 hours, and thawed, disintegrated by microtubule sliding when 1 mM MgATP was added. Microtubules and outer dense fibers (ODFs) were usually extruded in groups or "bundles". A total of 44.5% of the cells extruded two distinct bundles, one from each side of the connecting piece, exhibiting opposite curvatures. Only one bundle was observed in 46.2% of the cells, and 9.2% showed no signs of sliding. Transmission electron microscopy (T.E.M.) showed one group consisting of the 4,5 6,7 elements, with the 9,1,2 elements on the other side of the axoneme making up the other bundle. T.E.M. revealed that when only one side of the axoneme had extruded elements, they were always from the 4,5-6,7 group. The remainder of the axoneme (8,9,1,2,3 and the central pair) was left relatively intact, suggesting a difference in the sliding response of the nine pairs of axonemal microtubules. These results indicate a predisposition for sliding between elements 7 and 8 over that between doublets 2 and 3, perhaps due to a disparity in activation thresholds. Also, both Ni2+ and Cd2+ appear to selectively block activation of 2 3 interdoublet sliding. Incubation with 0.25 mM Ni2+ prior to adding MgATP modified the percentages of sliding patterns: 8.6% demonstrated two-sided extrusion, 58.2% showed one-sided, and 33.2% had no extruded bundles. Again, when half the axoneme was missing, it was always the 4,5-6,7 group. Ten micromolar Cd2+ altered the sliding pattern similarly to Ni2+, with 28% two-sided extrusion, 55.9% one-sided extrusion and 16.1% with no extruded bundles. Either pretreatment regimen impeded extrusion of the 9,1,2 group in a high percentage of cells, compared to untreated cells. This specific inhibition of the 9,1,2 side by Ni2+ or Cd2+ is especially significant since Ni2+ also inhibits spontaneous wave initiation in bull sperm (Lindemann et al.: Journal of Cell Biology 87:420-426, 1980), and both Ni2+ and Cd2+ reportedly block the flagellar Ca(2+)-response in rat sperm (Lindemann and Goltz: Cell Motility and the Cytoskeleton 10:420-431, 1988; Lindemann et al.: Cell Motility and the Cytoskeleton 20:316-324, 1991). PMID- 8221909 TI - Distribution of F-actin elongation sites in lysed polymorphonuclear leukocytes parallels the distribution of endogenous F-actin. AB - We compared, on lysed polymorphonuclear leukocytes (PMNs), the spatial distributions of sites that nucleate actin polymerization with the spatial distribution of endogenous F-actin. Sites nucleating polymerization of exogenous actin were detected by incubating lysed cells with rhodamine-labeled G-actin under polymerizing conditions. Endogenous F-actin was stabilized and stained by lysis of cells into fluorescein-labeled (FITC) phalloidin. We found the distributions of rhodamine and fluorescein intensities in a given cell, resting or stimulated with chemoattractant, to be similar. Thus, after lysis the number of sites able to nucleate actin polymerization is proportional to the local F actin concentration. Quantitative fluorescence microscopic analysis also demonstrated that (1) if cells were stimulated with chemoattractant shortly before lysis, the total fluorescence per cell of both fluorophores went up; (2) if peptide was diluted shortly before lysis, the endogenous F-actin in the lamellae was dramatically reduced, but nucleation sites persisted, giving a high rhodamine to fluorescein ratio; and (3) there was a small increase in the ratio of rhodamine (exogenously grown actin) to fluorescein (endogenous F-actin) in a region near the lamellar/endoplasm border, centripetal to regions of the highest concentration of endogenous F-actin. The rhodamine signal appeared to be due to in situ actin polymerization probably nucleated by existing free barbed ends, since (1) the rhodamine signal increased linearly with time with no detectable lag if the actin concentration was above that of the critical concentration of the barbed end; (2) the rhodamine signal was dramatically reduced if lysates were incubated with gelsolin-actin complex (which stably caps barbed ends), then washed before the rhodamine G-actin was added; and (3) the number of nucleation sites at the time of lysis is similar to the number of the barbed ends of actin filaments determined by the kinetics of depolymerization [Cano et al., 1991]. The fact that the distribution of exogenous actin polymerization paralleled the endogenous F-actin suggests that the number of free barbed ends per F-actin is roughly constant. If all filament ends were free, or if a constant fraction of the filaments ends were free, these data would suggest that the mean filament length is roughly constant throughout the cell. PMID- 8221910 TI - Peptidases in the CNS: formation of biologically active, receptor-specific peptide fragments. AB - Peptides function as chemical signals between cells of multicellular organisms, or different organisms, via specific receptors on target cells. Many hormones, neuromodulators, and growth factors are peptides. Because there is no known reuptake system for peptides at the nerve terminal, the biological activity of peptides in the extracellular space is regulated by enzymatic degradation and extracellular metabolism. For example, angiotensin I is processed extracellularly in the lung by angiotensin-converting enzyme (ACE; E.C. 3.4.15.1), a peptidyl dipeptidase, to form the potent vasoconstrictor hormone angiotensin II. When neuropeptides are released from neurons into the extracellular space, specific peptidases also can modulate the peptidergic signal by generating smaller, biologically active fragments via products with similar or dissimilar characteristics of the parent peptide. Therefore, receptor-binding selectivity of a released peptide hormone can be regulated by peptidases. Because peptidases may play a key role in the extracellular regulation of peptidergic signaling, alterations in peptidase activities by drugs or disease states may lead to disruptions in biological homeostasis. The subject of this article is the role of peptidases in the central nervous system in the formation of biologically active, receptor-specific peptides from peptide E, beta-endorphin, neurotensin, and cholecystokinin. PMID- 8221911 TI - Experimental neurobiology of central nervous system trauma. AB - Traumatic injuries to the brain or spinal cord cause irreversible tissue damage by at least three mechanisms: through consequences of mechanical disruption of neurons or their projections; through biochemical or metabolic changes that are initiated by the trauma; and through reactive inflammatory or gliotic changes. During the past decade, considerable data have been accumulated regarding cellular and biochemical events associated with posttraumatic tissue damage. This has led to the application of pharmacological strategies to limit secondary injury and subsequent neurological deficits. Such research has resulted in the first effective treatment of human spinal cord injury, with other promising treatments in late preclinical or early clinical development. PMID- 8221912 TI - Metabolic mapping with cellular resolution: c-fos vs. 2-deoxyglucose. AB - 2-Deoxyglucose (2DG) studies have been most useful in mapping activated regions of the nervous system. Cellular localization studies using 2DG have been less rewarding, but results are consistent with current views that increases of 2DG accumulation produced by synaptic activation represent increases in glycolytic glucose metabolism occurring mainly in presynaptic neuronal and possibly glial elements. Immediate early genes (IEGs), including the fos, jun, and NGFI-A families, are induced by a wide variety of intracellular signaling pathways. The nuclear localization of the protein products of these genes and their induction by a variety of stimuli make them useful in metabolic activation studies carried out at the cellular level. IEGs have been induced in neurons by osmotic, bacterial endotoxin, steroids, stress, and other hormonal stimuli; by light, auditory, painful, and other sensory stimuli; during stimulation of motor cortex and other motor behaviors; and by various drugs and toxins that act on a variety of neurotransmitter systems, including dopamine and glutamate. In addition, the localization of c-fos gene expression identifies cells that respond to growth factors in vivo. Retinal Muller cells, the major glial cell type of the retina, demonstrate nuclear Fos immunostaining after the intravitreal injection of epidermal growth factor (EGF) or transforming growth factor-alpha (TGF-alpha). This observation demonstrates that adult glia can respond to these growth factors in vivo. The investigation of early response gene expression may be particularly useful for elucidating the role of trophic factors in the cellular response to central nervous system injury. PMID- 8221913 TI - The neurobiology of late-life psychosis. AB - Developments in understanding the neurobiology of late-life psychosis have centered primarily on structural, functional, and neurochemical factors. Structural factors have included the study of white matter hyperintensity signals on magnetic resonance imaging of the brain, and associated vascular diseases as possible etiologies of psychosis in the elderly. Functional studies have continued to note frontal and medial temporal lobe dysfunction. Important neurochemical factors in psychosis include dopamine systems, serine metabolism, and the role of neurotensin. The neurochemical basis of psychosis has been proposed as a dysfunction of the corticostriatal pathways with negative feedback disinhibition of the thalamus with resultant sensory flooding, which leads to the perception of psychosis in susceptible patients. PMID- 8221914 TI - [Chorioamnionitis and perinatal infection in neonates]. AB - The authors investigated the relationship between histologically confirmed chorioamnionitis and the development of adnatal infection in the neonate. From a total of 4,144 deliveries during the investigation period-January 1, 1988 to December 31, 1989-252 placentas of neonates after pathological deliveries were examined. Chorioamnionitis was recorded in 28.6%, i.e. in 72 placentas inflammatory changes were found. Adnatal infections of neonates with chorioamnionitis were detected in 20.8%, i.e. 15 neonates. On the other hand, in the group of neonates where no inflammatory changes of the placenta were present adnatal infections were recorded in 8 infants, i.e. 4.4% (p = 0.00004). The mortality rate from adnatal infections in the group of neonates with chorioamnionitis was 4.16%. In the group of neonates without chorioamnionitis no death due to adnatal infection was recorded (p = 0.00000). Chorioamnionitis is thus associated with a significantly higher incidence of clinical adnatal infection in neonates and mortality due to this infection. PMID- 8221915 TI - [Early-stage lyme borreliosis during pregnancy: treatment in 15 women with erythema migrans]. AB - The authors treated and followed-up since July 1987 9 women with non-complicated erythema migrans and 4 women with erythema migrans associated with general signs and symptoms during pregnancy; two patients were treated only after delivery. For treatment antibiotics of the penicillin series were used. The course of gestation in 14 patients was without complications, one patient was followed-up on account of oedema of the lower extremities. The infants of 14 women were born in term, one infant was born during the 37th week of gestation. All infants were eutrophic. Umbilical blood for serological examination was collected from 8 infants, elevated IgG antibody levels against Borrelia burgdorferi were detected in one. The post-partum development of all infants was normal. During the subsequent follow-up for 6-26 months elevated antibodies against Borrelia burgdorferi were not found in any of the 4 infants subjected to serological examination. One child was operated at the age of one year on account of a patent ductus arteriosus, another child was treated at the age of two years on account of cryptorchism, two children were treated on account of hypoplastic enamel, in one infant at the age of 10 months the psychomotor development was retarded by two months. Transplacental transmission of Borrelia burgdorferi was not proved. With regard to the variety of the observed deviations and the small number of data the problem of a possible teratogenic action of this spirochete remains open. PMID- 8221917 TI - [Therapeutic strategy in cervical carcinoma in young women]. PMID- 8221916 TI - [Sexual development in adolescent girls]. AB - The authors investigated by interviews in Frantiskovy Lazne the sexual development and sex life in 614 Czech girls aged 16 to 18 years. It was revealed that the sexual development in 311 girls, where in the course of the year before convalescent treatment in Frantiskovy Lazne appendectomy was performed and in 89 probands who were operated twice or more frequently, was highly significantly retarded as compared with 163 probands who had no surgery so far. Comparison of the findings in 426 girls from complete and emotionally balanced families and 140 probands of divorced parents did not reveal a significant difference in their sexual development. PMID- 8221918 TI - [Fermalac Vaginal (Rougier Inc.) in the prevention of colpitis in pregnancy]. PMID- 8221919 TI - [Donor insemination in a non-government health care setting]. PMID- 8221920 TI - [Benign cystic mesothelioma of the peritoneum]. PMID- 8221921 TI - [Endometrial protein PP14, its structure, function and clinical use]. PMID- 8221922 TI - [Epilepsy in pregnancy]. PMID- 8221923 TI - [New indications for mifepristone (RU 486), an antiprogesterone]. PMID- 8221924 TI - [Errors and omissions in cytodiagnosis]. PMID- 8221925 TI - MCB elements and the regulation of DNA replication genes in yeast. AB - In eukaryotic organisms, genes involved in DNA replication are often subject to some form of cell cycle control. In the yeast Saccharomyces cerevisiae, most of the DNA replication genes that have been characterized to date are regulated at the transcriptional level during G1 to S phase transition. A cis-acting element termed the MluI cell cycle box (or MCB) conveys this pattern of regulation and is common among more than 20 genes involved in DNA synthesis and repair. Recent findings indicate that the MCB element is well conserved among fungi and may play a role in controlling entry into the cell division cycle. It is evident from studies in higher systems, however, that transcriptional regulation is not the only form of control that governs the cell-cycle-dependent expression of DNA replication genes. Moreover, it is unclear why this general pattern of regulation exists for so many of these genes in various eukaryotic systems. This review summarizes recent studies of the MCB element in yeast and briefly discusses the purpose of regulating DNA replication genes in the eukaryotic cell cycle. PMID- 8221926 TI - Isolation of the TYE2 gene reveals its identity to SWI3 encoding a general transcription factor in Saccharomyces cerevisiae. AB - The TYE2 gene was identified by recessive mutations which result in a significant reduction of Ty-mediated ADH2 expression. We cloned the TYE2 gene and analyzed its sequence. A large open reading frame of 825 codons was found encoding a rather hydrophilic, 93-kilodalton protein which contains a highly acidic region at its N-terminus. By sequence comparison we found that TYE2 is identical to gene SWI3 which has recently been shown to encode a nuclear protein which may function as a global transcription activation factor. The TYE2/SWI3 protein is necessary for the initiation of Ty1 transcription at its major initiation site in the delta element. Furthermore TYE2 function seems to be important for the expression of a variety of Ty-unrelated functions such as ADH1 expression, sporulation, growth on maltose, galactose, raffinose, and on non-fermentable carbon sources. PMID- 8221927 TI - DNA polymerase III is required for DNA repair in Saccharomyces cerevisiae. AB - We have studied the role of DNA polymerase III, encoded in S. cerevisiae by the CDC2 gene, in the repair of yeast nuclear DNA. It was found that the repair of MMS-induced single-strand breaks is defective in the DNA polymerase III temperature-sensitive mutant cdc2-1 at the restrictive temperature (37 degrees C), but is not affected at the permissive temperature (23 degrees C). Under conditions where only a small number of lesions was introduced into DNA (80% survival), the repair of MMS-induced damage could also be observed in the mutant at the restrictive temperature, although with low efficiency. When the quantity of lesions increased (50% survival or less), the repair of single-strand breaks was blocked. At the same time we observed a high rate of reversion in the meth, his and trp loci of the cdc2-1 mutant under restrictive conditions. The results presented suggest that DNA polymerase III is involved in the repair of MMS induced lesions in yeast DNA and that the cdc2-1 mutation affects the proofreading activity of this polymerase. PMID- 8221928 TI - Cloning and sequence analysis of the glucoamylase gene of Neurospora crassa. AB - A 1.0-kb DNA fragment, corresponding to an internal region of the Neurospora crassa glucoamylase gene, gla-1, was generated from genomic DNA by the polymerase chain reaction, using oligonucleotide primers which had been deduced from the known N-terminal amino-acid sequence or from consensus regions within the aligned amino-acid sequences of other fungal glucoamylases. The fragment was used to screen an N. crassa genomic DNA library. One clone contained the gene together with flanking regions and its sequence was determined. The gene was found to code for a preproprotein of 626 amino acids, 35 of which constitute a signal and propeptide region. The protein and the gene are compared with corresponding sequences in other fungi. PMID- 8221929 TI - Amino-acid substitutions in the zinc finger of NIT2, the nitrogen regulatory protein of Neurospora crassa, alter promoter element recognition. AB - NIT2, the major nitrogen regulatory protein of Neurospora crassa mediates nitrogen catabolite derepression of the structural genes which specify enzymes of nitrogen catabolism. The promoter of the structural gene for L-amino acid oxidase, a nitrogen-regulated enzyme, was found to contain two NIT2 binding sites, each with two copies of a GATA core consensus sequence. Site-directed mutagenesis was employed to create amino-acid substitutions within the single zinc-finger region of NIT2, which serves as the DNA-binding domain. The affect of those mutations upon NIT2 function in vivo in the activation of three separate structural genes was examined by transformation assays and relevant enzyme activities, and DNA-binding activity in vitro was determined by gel band mobility shift assays. It was shown that specific amino-acid residues within the zinc finger loop region of NIT2 are important for DNA-binding activity, whereas other residues influence the specificity of DNA binding. Mutant NIT2 proteins were obtained which retain DNA-binding activity and alter the specificity of DNA recognition, thus allowing a distinction between related DNA elements. PMID- 8221930 TI - Assignment of linkage groups to the electrophoretically-separated chromosomes of the fungus Podospora anserina. AB - An electrophoretic karyotype of the filamentous fungus Podospora anserina has been obtained using contour-clamped homogeneous electric field gel electrophoresis. Six chromosomal bands were separated with one migrating as a doublet. The size of the chromosomes was estimated to be between 3.8 and 6.0 megabase pairs (mb) using the chromosomes of Schizosaccharomyces pombe as size standards, giving a total genome size of about 34 mb for the P. anserina genome. Homologous probes were used to assign five of the seven linkage groups (LGs) to chromosomal bands on the gel. Analysis of reciprocal translocation strains allowed us to complete the karyotype. In decreasing size order, the P. anserina chromosomes are LG I (6.0 mb); LG II (5.5 mb); LG V (5.1 mb); LG III (4.9 mb); LGs VI and VII (4.3 mb) and LG IV (3.8 mb). PMID- 8221931 TI - Transformation of Trichoderma reesei with the Hormoconis resinae glucoamylase P (gamP) gene: production of a heterologous glucoamylase by Trichoderma reesei. AB - A cDNA encoding for the glucoamylase P enzyme (GAMP) of the fungus Hormoconis resinae was introduced into the cellulolytic filamentous fungus Trichoderma reesei under the control of the promoter of the major cellulase gene (cbh1) of Trichoderma. The transforming vector plasmid used was found to be integrated into the genome of T. reesei at various locations and in multiple copies. The size of the GAMP secreted by Trichoderma varied because of different glycosylation patterns. The best transformant strains secreted about 700 mg/l of active GAMP, which is 20-fold more than obtained with H. resinae. PMID- 8221932 TI - Structure, expression, and phylogenetic relationships of a family of ypt genes encoding small G-proteins in the green alga Volvox carteri. AB - In addition to the previously described gene yptV1 encoding a small G-protein we have now identified and sequenced four more ras-related ypt genes (yptV2-yptV5) from the green alga Volvox carteri. The four new genes encode polypeptides consisting of 203 to 217 amino-acid residues that contain the typical sequence elements (GTP-binding domains, effector domain) of the ypt/rab subgroup of the Ras superfamily. Comparison of the derived amino-acid sequences from the V. carteri ypt gene products and their Ypt homologs from other species revealed similarity values ranging from 60% to 85%, whereas intraspecies similarities were found to approach only 55%. The coding sequences are interrupted by 5-7 introns of variable size (70-1000 nucleotides) occupying different positions in the genes. Reverse-transcribed samples of stage-specific RNAs were PCR-amplified with primers specific to yptV1, yptV3, yptV4, and yptV5 to determine if yptV transcription might be restricted to either cell type or to a specific stage of the life cycle. These experiments demonstrated that each of these genes is expressed throughout the entire Volvox life cycle and in both the somatic and the reproductive cells of the alga. The transcription start sites of yptV1 and yptV5 were mapped by primer extension. Expression of recombinant yptV cDNA in E. coli yielded recombinant proteins that bound GTP specifically, demonstrating a property which is typical for small G-proteins. The derived YptV polypeptide sequences were used to group them into four distinct classes of Ras-like proteins. These are the first proteins of the Ras superfamily to be identified in a green alga. We discuss the possible role of the YptV-proteins in the intracellular vesicle transport of Volvox. PMID- 8221933 TI - Mitochondrial DNA of Chlamydomonas reinhardtii: the structure of the ends of the linear 15.8-kb genome suggests mechanisms for DNA replication. AB - The mitochondrial genome of Chlamydomonas reinhardtii is a linear double-stranded DNA of 15.8 kb. With the exception of the termini its DNA sequence has been published. Here we describe the unique structure of the two termini determined from cloned fragments or, for the very terminal sequences, by the Maxam and Gilbert method after 5' labeling of uncloned terminal fragments. The 15.8-kb DNA is characterized by terminal inverted repeats of 531 or 532 bp in length including long 3' extensions. The 3' single-stranded extensions of the left and right ends are non-complementary, identical in sequence, and comprise 39 to 41 nucleotides. Remarkably, the linear genome possesses in addition an internal 86 bp repeat of the two outermost sequences. The unusual structure of the 15.8-kb DNA termini is compared with those of other linear mitochondrial DNAs. Possible mechanisms of 15.8-kb DNA replication are discussed. PMID- 8221934 TI - A gene involved in the biogenesis of c-type cytochromes is co-transcribed with a ribosomal protein gene in wheat mitochondria [corrected]. AB - Sequence analysis of a transcribed region of the wheat mitochondrial (mt) genome revealed two open reading frames (orfs) coding for proteins of 589 and 174 amino acids. Both genes are co-transcribed in a 2.6-kb RNA. The largest orf codes for a hydrophobic protein which bears similarity to a bacterial protein involved in the biogenesis of c-type cytochromes. Its corresponding RNA sequence is fully edited at 34 positions. The second orf encodes a protein homologous to the amino terminal third of E. coli ribosomal protein S1, corresponding to the ribosome binding domain of this protein. Its RNA sequence is edited at four positions, one of the edits creating a stop codon. The presence of both proteins in wheat mitochondria was demonstrated using specific antibodies raised against fusion proteins obtained in E. coli from the corresponding cDNAs. PMID- 8221935 TI - The presequence of cytochrome c1 from potato mitochondria is encoded on four exons. AB - The structural organization of a nuclear gene encoding cytochrome c1 from potato was determined. The gene spans 5.1 kb and contains eight introns. All intron/exon junctions follow the GT/AG rule. Functional domains of the mature cytochrome c1 protein are located on separate exons. The presequence, which targets the cytochrome c1 precursor to the mitochondrion and to the correct intra mitochondrial location, is encoded on the first four exons. The largest intron (2.8 kb) separates the information for mitochondrial targeting from the "intra mitochondrial sorting domain" of the cytochrome c1 protein. In contrast to other organellar precursor proteins, there is no intron between the DNA sequence encoding the presequence and the mature protein. This may indicate that during evolution the genetic information for the prokaryotic cytochrome c1 was transferred to the nucleus together with the bacterial secretion signal which is structurally and functionally related to "intramitochondrial sorting domains". PMID- 8221936 TI - The chloroplast and mitochondrial DNA type are correlated with the nuclear composition of somatic hybrid calli of Solanum tuberosum and Nicotiana plumbaginifolia. AB - This paper describes the analysis of chloroplast (cp) DNA and mitochondrial (mt) DNA in 21 somatic hybrid calli of Solanum tuberosum and Nicotiana plumbaginifolia by means of Southern-blot hybridization. Each of these calli contained only one type of cpDNA; 14 had the N. plumbaginifolia (Np) type and seven the S. tuberosum (St) type. N. plumbaginifolia cpDNA was present in hybrids previously shown to contain predominantly N. plumbaginifolia chromosomes whereas hybrids in which S. tuberosum chromosomes predominated possessed cpDNA from potato. We have analyzed the mtDNA of these 21 somatic hybrid calli using four restriction enzyme/probe combinations. Most fusion products had only, or mostly, mtDNA fragments from the parent that predominated in the nucleus. The hybrids containing mtDNA fragments from only one parent (and new fragments) also possessed chloroplasts from the same species. The results suggest the existence of a strong nucleo-cytoplasmic incongruity which affects the genome composition of somatic hybrids between distantly related species. PMID- 8221937 TI - Multicopy SUP35 gene induces de-novo appearance of psi-like factors in the yeast Saccharomyces cerevisiae. AB - Previously, we have shown that plasmid-mediated multiplication of Saccharomyces cerevisiae wild-type SUP35 gene leads to omnipotent suppression and is incompatible with psi-factor, which is an endogenous extrachromosomal suppressor. Here, we describe a frequent de-novo appearance of psi-like factors in mitotic progeny of yeast transformants containing multicopy SUP35 gene. PMID- 8221938 TI - Effective long range mapping in Schizosaccharomyces pombe with the help of swi5. AB - The switching gene swi5 has a function in mating-type switching. In addition, the swi5 mutation causes an increased radiation sensitivity and reduces meiotic recombination about ten-fold. Based on the latter property, an experimental protocol was developed for using swi5 in long-range mapping in S. pombe. It is suitable for a speedy mapping of any new gene which has not yet been cloned. The procedure was used to clarify the map positions of some genes. PMID- 8221939 TI - The isolation and nucleotide sequence of the pyruvate decarboxylase gene from Kluyveromyces marxianus. AB - We have determined the nucleotide sequence of a 2360-basepair (bp) region of the Kluyveromyces marxianus genome containing the structural gene for the enzyme pyruvate decarboxylase (PDC). Comparison of the deduced amino-acid sequence of this gene to that of the Saccharomyces cerevisiae PDC genes reveals extensive homology including a motif common to thiamin diphosphate-dependent enzymes. PMID- 8221940 TI - [Eye manifestations of hypovitaminosis A]. AB - Two cases of a heavy hypovitaminosis A (tumor of the pancreas and an insufficient resorption accompanying the syndrome of a short intestine) are presented. The eye symptomatology consisted of lowered dark-adaption curve, absent electrical reactivity and lowered contrast sensitivity. After restoring the normal vitamin A level in the blood the eye symptoms vanished. PMID- 8221941 TI - [Visual evoked potentials in reversal and motion-onset stimulation of visual fields in patients with retrobulbar neuritis]. AB - In 21 patients with unilateral retrobulbar neuritis the pattern reversal evoked potentials and motion-onset visual potentials were investigated with the aim to find out whether there is some difference in affection of pattern detecting (parvocellular) and motion detecting (magnocellular) visual pathways in this disease. The pattern reversal potentials were found to be abnormal in all 21 patients: they showed either significantly prolonged latencies and reduced amplitudes of the main positive P100 peak or no clear response (in one patient) upon the stimulation of the eye with neuritis. In contrast, the main negative peak of the motion-onset potentials was significantly delayed only in 15 patients and its amplitude was distinctly reduced only in 3 patients. The results indicate that in retrobulbar neuritis the pattern detecting nerve fibres from the central part of the retina (tested by reversal potentials) are affected earlier and to a greater extent than the more peripherally distributed motion detecting fibres (tested by motion-onset potentials). PMID- 8221942 TI - [18-year survival in a case of repeated excision of malignant conjunctival melanoma]. AB - The case of 18 years survival with repeatedly excised conjunctival malignant melanoma is described. The survival period is extraordinary long in that case. However, the case is illustrative for some general characteristics of conjunctival melanoma, particularly in a comparison with cutaneous melanomas. The importance of regular following of patients with primary acquired melanosis is stressed in conclusion. PMID- 8221943 TI - [Tumors of the inner angle of the eye]. AB - The authors present an account of their longstanding experience with treatment of tumours affections of the inner angle of the eye. Careful preoperative examination incl. assessment of possible obstruction of the lacrimal pathways and CT examination of the affected area, which was the only method which did not produce falsely negative results, is essential. In this area they recommend radical surgery of tumours infiltrating into deep layers, i.e. extensive excision, incl. some sound tissue, and covering of the defect by a frontal flap. With regard to the danger of inadequate freezing they do not recommend in this area cryodestruction in deep tumours. The authors draw attention to the necessity of close collaboration of the ophthalmologist and otorhinolaryngologist which makes it possible to supplement the operation, when the paranasal sinuses are affected, which was essential in the great majority of the authors' patients. PMID- 8221944 TI - [Ostensible ultrasonic changes after internal tamponade of the retina with silicone oil]. AB - Echographic control after pars plana vitrectomy (PPV) and internal tamponade of the retina with silikon oil (SO) is necessary in case of opaque optical media (bulous keratopathy, bleeding and exsudation within anterior chamber, complicated cataract). In interpretation of echographic image some difficulties or errors may be caused by physical properties of SO, as for example reduced speed of propagation of ultrasound in this media, ultrasound absorption coefficient, acoustic impedance and reflection coefficient. PMID- 8221945 TI - [A method for opening the anterior chamber in capsulorhexis in cataract surgery via the scleral tunnel]. PMID- 8221946 TI - [Isovolemic hemodilution--one of the possibilities in therapy of retinal vein occlusion]. AB - The author submits a review of findings on the basis of which isovolaemic haemodilution is recommended as a therapeutic procedure in occlusion of retinal veins. By modification of the viscosity of blood rheological conditions change and thus the conditions for repair of the affected retina improve. The method of the procedure is described. The author mentions also indications and contraindications of this procedure and specifies the group of patients indicated for this treatment. The author considers the presented review an adequate basis for using haemodilution in other departments, similarly as in the author's, where it has been used now for more than three years. PMID- 8221947 TI - [Quadrant cyclocryocoagulation. Initial experience]. AB - The authors describe a modification of cycloccryocoagulation /CCC/ they used in the treatment of different types of glaucoma in a group of 320 eyes. Cryodestruction of the ciliary body was implemented with a probe at a temperature of -60 degrees C for a period of 2 x 20sec. in two points of opposite qaudrants, using a transconjunctival approach. The probe was applied at a distance of 3 mm from the limbus. Following a period of 12 months to 3 years after operation they investigated the intraocular pressure and visual acuity of the treated eyes. They recorded a hypotonic effect comparable to cryocoagulation performed by the classical procedure; the incidence of complications was lower. PMID- 8221948 TI - [Diode lasers in ophthalmology]. AB - The authors present an account of diode lasers based on publications in the literature and their initial own experience. A brief physical characteristic of diode lasers is given. PMID- 8221949 TI - [Secondary uveitis in intraocular lenses]. PMID- 8221950 TI - [The effect of antimetabolites on healing in anti-glaucoma surgery]. PMID- 8221951 TI - [Bronchogenic carcinoma accompanied with gynecomastia. A case report]. AB - A case of bronchogenic carcinoma accompanied with gynecomastia was reported. The enlargement of bilateral breasts was discovered for 3 months before admission with continuous low grade fever, dyspnea and weight loss. The patient was a heavy smoker and had chronic bronchitis. Physical examination revealed bilateral enlargement of breasts, the diameter of each breast was 12 cm and the height was 6 cm. Serum human chorionic gonadotropin (HCG) was > 2000 miu/ml, prolactin (PRL) 41.5 ng/ml, luteinizing hormone (LH) > 200 miu/ml. The chest X-ray film showed scattered nodular shadows in both lung fields. Tumor cells were found in the lung tissue obtained by biopsy after death suggesting of adenocarcinoma. The pathogenesis of gynecomastia was discussed. PMID- 8221952 TI - [The current status of tuberculosis and the advances in its treatment]. PMID- 8221953 TI - [A study on the relapse and its potential causes in pulmonary tuberculosis]. AB - A retrospective cohort study was conducted to acquire the general picture of relapse in pulmonary tuberculosis in China and to clarify the relationship between potential causes and relapse rate. All data were collected from 4,797 initial sputum-positive cases. Results showed: (1) Bacteriological relapse rate within two years after chemotherapy was 4.6%. (2) Cumulative relapse rates within half a year and a year only accounted for respectively 28% and 43% of that within two years after chemotherapy. (3) Patients over 60 years old, with drug-resistant bacilli, with long excretion of bacilli, or with residual cavity(ies) or bacterial-negative conversion of less than 3 months at the end of treatment, were the high risk population of relapse. PMID- 8221954 TI - [The diagnosis of tuberculous meningitis using polymerase chain reaction]. AB - The PCR system can detect 10 pg of DNA and 10-50 cfu of M. tuberculosis. The sensitivity and specificity rate of our PCR system about 80 cases of tuberculosis meningitis and 63 cases of nontuberculous meningitis is 85% and 98.4%, respectively. Comparing with the routine mycobacterial examination in 61 cases of tuberculous meningitis, the positive rate of PCR is 86.9%, while the positive rate of routine mycobacterial examination (smear and/or culture) is only 13.1%. In the 53 negative cases of routine mycobacterial examination, the positive rate of PCR can reach 86.8%. The whole detection can be finished within 2 days. PMID- 8221955 TI - [Discussion on the relationship between trace elements in cerebrospinal fluid in tuberculous meningitis]. AB - The concentration of Mg, Zn, Cu, Fe, Mn and V in the CSF of 38 cases of tuberculous (tbc) meningitis and those in 40 cases of normal control were measured. Results showed that concentration of Mg, Zn or Fe in tested group was higher and that of Cu was lower than that in control group, but there was no significant difference between Mn and V's concentration in 2 groups. The authors suggested that those findings may be helpful in improving the treatment of tbc meningitis. PMID- 8221956 TI - [Clinical significance of soluble interleukin 2 receptor alpha in the pleural fluid]. AB - The levels of soluble interleukin 2 receptor alpha (sIL-2R alpha) were measured in pleural fluid from 38 patients including 16 cases with tuberculous pleurisy, 14 with malignant pleural effusions and 8 with transudative effusions by using ELISA technique. The level of sIL-2R alpha in tuberculous pleurisy is markedly higher than in malignant pleural effusions (P < 0.01). The levels of sIL-2R alpha in tuberculous pleurisy and in malignant pleural effusions are significantly higher than in transudative effusions (P < 0.01). The clinical significance is discussed. PMID- 8221957 TI - [Symptomatic survey on pulmonary tuberculosis in Shanghai rural areas]. AB - A study on the discovery of pulmonary tuberculous patients in 72 villages 312,092 population in 9 counties of Shanghai rural areas by symptomatic survey was reported. The survey showed that only 8.21% of the population (25631 cases) had suspected TB symptoms, the discovery rate of > 45 age-group was still around 10%, 233 cases of active PUL TB were discovered after all the suspected subjects were examined, and 71 cases of sputum (+) were identified. For the > 60 age-group the discovery rate of active PUL TB was 64.8% (151/233) and the sputum (+) cases was 66.2% (47/71). PMID- 8221958 TI - [Assessment on the epidemiological efficiency and treatment of smear positive pulmonary tuberculosis]. AB - An epidemiological project for discovery and treatment of sputum (+) pulmonary tuberculous patients had been carried out during 1988-1990 in patients collected, this number was 3.1 times as the number obtained in the five year pre-application time. The annual mean value of proportion of smear (+) cases of newly registered patients was 63.53%, the average year rate of treatment coverage was 96.2%, the year complication rate of treatment was 89.2%, the sputum negative conversion rate was 87.0%, the cure rate in cohort analysis of management on chemotherapy was 85.8% in the period of 1988-1990. The annual reduction rate of prevalence rate of sputum (+) cases increased to 8.9% annually, but before this program (1979-1985), the percentage increased 1% per year. Thus, the epidemiological efficacy is significant. PMID- 8221959 TI - [Study on quality of diagnosis in patients with sputum negative pulmonary tuberculosis in rural area]. AB - 1194 cases (3.9%) were randomly selected from all newly registered patients with sputum negative pulmonary tuberculosis (PTB) of Jiangsu Province in 1991. After reevaluation by specialists, final diagnosis was made. 94 cases (7.9%) were excluded because of insufficient information, the diagnosis of PTB for 107 patients (9.7%) of the remained 1100 cases were not correct. Results indicate that one of the causes for the low sputum positive rate of newly registered patients with PTB was probably due to the comparatively large proportion of misdiagnosed sputum negative patients. PMID- 8221960 TI - [Changes in interleukin-1 released by pulmonary alveolar macrophage in patients with interstitial lung disease]. AB - To evaluate the activity of PAM, levels of IL-1 released by PAM in patients with ILD (non smokers) were measured using lipopolysaccharide (LPS) stimulation and thymocyte proliferation method, healthy non-smokers as control group. The results showed that IL-1 released by PAM in patient groups both with and without LPS stimulation were significantly higher than that in control group, and also IL-1 released by PAM in healthy smokers was significantly higher than that in health non-smokers. It was indicated that IL-1 might play an important role in the process of ILD and chronic pulmonary inflammation. PMID- 8221961 TI - [Effects of tetrandrine on pulmonary hypertension in patients with chronic obstructive pulmonary disease]. AB - Tetrandrine were given to nine patients with pulmonary hypertension secondary to chronic obstructive pulmonary disease and the effects of central hemodynamics and blood gases were studied. 60 minutes after the administration of the medicine, total pulmonary vascular resistance (TPVR) and pulmonary artery mean pressure (mPAP) were reduced by 23% (from 59.2 KPa.s/L to 45.6k Pa.s/L (1 kPa = 7.5 mmHg, P < 0.05) and 15% (from 4.0 kPa to 3.4 kPa, P < 0.05) respectively. Systemic artery mean pressure (mSAP) only decreased by 4.9%. There were no significant differences between the values of cardiac output (co), PaO2, SaO2 before and after the administration of tetrandrine. PMID- 8221962 TI - [The study on prethrombotic state in patients with chronic obstructive pulmonary disease and cor pulmonale]. AB - We measured PT, TT, KPTT, Fg, vWF, AT-IIIAg, AT-IIIA, alpha 2M, TXB2, 6-keto-PGF1 alpha, PLg, tPAAg, tPAA and PAI of patients with acute onset of chronic bronchitis, pulmonary emphysema, and cor pulmonale. The results were that many above parameter had a worsening tendency along with deterioration of COPD, and Fg, vWF, TXB2, 6-keto-PGF1 alpha, tPAA and PAI were more sensitive than others. The analysis of multiple liner regression of 22 blood items in patients with cor pulmonale showed that pH, PaO2, PaCO2 had correlation with many items of prethrombotic state. We consider that in the progress of COPD, prethrombotic state gradually appeared and aggravated. The causes may be related with repeatedly infections, low O2, high CO2 and imbalance of acid-base equilibrium. PMID- 8221963 TI - [Application of bedside monitoring with arterized capillary blood, transcutaneous and end tidal carbon dioxide during mechanical ventilation]. AB - Arterial blood, arterialized capillary blood, transcutaneous, end tidal carbon dioxide pressure (PaCO2, PcCO2, PtcCO2, PetCO2) and physiological respiratory deadspace to tidal volume ratio (VD/VT) were observed in 12 patients with chronic obstructive pulmonary disease (group A), 17 postoperative patients of cardiac surgery (group B), and 16 healthy subjects (group C) under three mechanical ventilatory patterns. PcCO2, PtcCO2 and PetCO2 were found to correlate well with PaCO2 (r = 0.99, 0.97 and 0.88 respectively); P(a-et) CO2 correlated with age (year) in group B and C (r = 0.77 and 0.76), and all P(a-et) CO2 measurements of each of three groups highly correlated with VD/VT (r = 0.96 0.91 and 0.87 for group A,B,C respectively). The results suggested that the trend of PaCO2 variation can be deduced from PcCO2, PtcCO2 and PetCO2 with fair accuracy under various ventilatory patterns in subjects with stable circulatory function, and P(a-et) CO2 is a simple quantitative index for evaluating physiological deadspace. PMID- 8221964 TI - Dihydrodiol dehydrogenase and its role in polycyclic aromatic hydrocarbon metabolism. AB - Dihydrodiol dehydrogenase(s) (DD) have been implicated in the detoxication of proximate (trans-dihydrodiol) and ultimate carcinogenic (anti-diol-epoxide) metabolites of polycyclic aromatic hydrocarbons (PAHs). These activities are catalyzed by soluble hydroxysteroid dehydrogenases and/or by aldehyde reductases. Molecular cloning indicates tha these enzymes have a high degree of sequence identity with members of the aldo-keto reductase super family. Substrate specificity studies indicate that non-K-region trans-dihydrodiols are the preferred substrates and that anti-dio-epoxides are not oxidized by the enzyme. The products of the DD reaction are transient catechols which auto-oxidize to PAH o-quinones. As a consequence of this auto-oxidation superoxide anion, hydrogen peroxide and semiquinone radicals are generated. Studies on the biotransformation of (+/-)-trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene indicate that in subcellular fractions from uninduced rat liver, DD plays a significant role in the metabolism of this proximate carcinogen. Thus, the formation of benzo[a]pyrene-7,8-dione is only superseded by the formation of tetraols which are derived from the anti-diol epoxide of benzo[a]pyrene [anti-BPDE;(+/-)-anti-7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene]. PAH-o-quinones produced by DD can inactivate the enzyme. These PAH-o-quinones also vary in their reactivity towards cellular nucleophiles, their cytotoxicity and their genotoxicity. Non-bay region and methylated bay-region PAH-o-quinones generated by DD are the most reactive Michael acceptors, and are also the most cytotoxic in hepatoma cells. Cytotoxicity results from the 1e- redox-cycling of the PAH-o-quinone, concomittant production of superoxide anion and a subsequent alteration in redoxstate. PAH-o-quinones are also genotoxic thus [3H]-benzo[a]pyrene-7,8-dione readily forms deoxyguanosine-adducts with native calf-thymus DNA, i.e., to the same extent as anti-BPDE. The cytotoxic and genotoxic properties of PAH-o quinones suggest that DD may initiate a hitherto unrecognized pathway of PAH activation. PMID- 8221965 TI - The effects of various inhibitors on the regulation of orotic acid excretion in sparse-fur mutant mice (spf/Y) deficient in ornithine transcarbamylase. AB - Experiments were conducted to determine whether the excessive orotic aciduria, induced in sparse-fur male mice (spf/Y) deficient in ornithine transcarbamylase (OTC), may be regulated by some inhibitors, such as acivicin (0.014 mmol/100 g body weight, i.p.), N-(phosphonoacetyl)-L-aspartate (PALA, 2.5 mg/100 g body weight, i.p.), adenine (3 g/kg diet) and cycloheximide (0.35 mmol/kg body weight, i.p.). We also administered ornithine (1 mmol/100 g body weight, i.p.), a substrate of the urea cycle, to alleviate the metabolic deficiency of arginine in spf/Y mice which may also be responsible for excessive orotic aciduria. The orotic aciduria remained insensitive to acivicin, indicating mitochondria as the source of carbamyl phosphate. However, orotate excretion was significantly decreased by PALA (P < 0.01), due to its effect on the aspartate transcarbamylase activity. The ingestion of adenine resulted in an increase (P < 0.05) of urinary orotate, suggesting the blockage of the utilization of orotate for nucleotide biosynthesis. Ornithine administration led to a reduction (P < 0.01) of the excretion of orotate induced by the OTC deficiency in these mice, indicating that one of the regulatory steps in its synthesis may be the availability of ornithine. There were no changes in urinary orotate excretion in spf/Y mice when treated with cycloheximide. On the other hand, pretreatment with cycloheximide in an artificial model of OTC deficiency (Swiss-ICR normal mice on an arginine deficient diet treated thereafter with norvaline, an inhibitor of OTC), caused a significant decrease in urinary orotate. These results suggest that spf/Y mice are unique in that the increased synthesis of orotate is not sensitive to cycloheximide. Perhaps this may reflect an adaptive phenomenon developed by the mutant mice to handle excess mitochondrial carbamyl phosphate and orotic acid. PMID- 8221966 TI - Alkylation of DNA by melphalan with special reference to adenine derivatives and adenine-guanine cross-linking. AB - Alkylation of DNA by melphalan gave four principal products, derived by mono alkylation of adenine at N-3 and guanine at N-7, and by cross-linking of adenine N-3 to guanine N-7, or of guanine N-7 to guanine N-7. Adenine-guanine cross linking was unexpected because the two principal nucleophilic centres, N-7 of guanine and N-3 of adenine, are situated in the 'wide' and 'narrow' grooves of the DNA double helix, respectively. These products could be isolated by their hydrolysis from DNA at neutral pH, followed by chromatography of the hydrolysate in an ion-pair ODS system using a solvent containing tetrabutylammonium hydroxide as ion-pairing reagent; this gave better separation than the previously described method using SP-Sephadex. PMID- 8221967 TI - Binding of Pt-pentamidine to nucleosomal DNA. Studies of the antiproliferative activity of the drug against human cancer cells. AB - In the present paper we present data showing that the effect of the binding of the antitumour drug Pt-pentamidine to nucleosomal DNA is the opposite to that of the cis-DDP compound since it causes strong stabilization of the double helix to heat denaturation and because in nucleosome:Pt-pentamidine complexes the nucleosomal denatured DNA is able to reassociate at 71 degrees C. Upon binding, the pentamidine ligand, by itself, also produces stabilization of the nucleosomal DNA but the effect is lower than that induced by Pt-pentamidine. It seems that in Pt-pentamidine:nucleosome complexes about 50% of the adducts are formed during the first hour of incubation of the nucleosomes with the drug since the increase in Tm of the DNA of these complexes is 53% of the total increase in Tm of the DNA of the Pt-pentamidine:nucleosome complexes formed in 48 h. The 'in vitro' screening of the antiproliferative activity of Pt-pentamidine against 60 tumour cell lines indicated that this antitumour compound shows higher antiproliferative activity against small cell lung, non-small cell lung and melanoma cancer lines than against the rest of the cell lines. PMID- 8221968 TI - Diminished drug metabolism due to hormonal effects of cobalt mesoporphyrin on flavoproteins. AB - Substituted metalloporphyrins, in addition to their use as pharmacological agents, are used to investigate metabolic pathways by inhibiting cytochrome P 450. We have examined the specificity of this approach with cobalt mesoporphyrin (CoMP). In vivo, CoMP (50 mumol/kg, s.c.) decreased rat hepatic microsomal cytochrome P-450, NADPH cytochrome P-450 reductase, benzphetamine N-demethylase (BZPH) activity, and thyroid hormones by > 50%, all of which returned to control levels after 45 days; testosterone levels were also reduced at this dose. The half-life of CoMP was 18 days, which is consistent with this sustained effect. At 10 mumol/kg of CoMP, the reductase activity was decreased, but cytochrome P-450 was unchanged. An effect of residual CoMP on the reductase was ruled out as the CoMP content of tissue fractions was not high enough to inhibit directly the reductase activity (even after 50 mumol CoMP/kg). However, immunoblots indicated a lower level of immunoreactive reductase protein following treatment. After 8 weekly doses of 1 mumol CoMP/kg, BZPH activity was 39% less than control but neither P-450 content nor reductase activity was significantly changed. The P-450 content and reductase activity in rabbits were much less affected by CoMP, perhaps due to differences in the disposition of CoMP. Thyroidectomy decreased reductase activity in rats to an extent that was seen with CoMP at 50 mumol/kg; CoMP treatment of thyroidectomised rats did not further decrease reductase activity. Supplementation with thyroid hormone blocked the CoMP-related decrease. The flavin-containing monooxygenase was decreased by CoMP and by castration, and the decrease was not blocked by the thyroid hormone supplement. Thus in the rat, the CoMP-related decreases in thyroid hormone and testosterone decrease flavoproteins that support or mediate monooxygenase activities. This is contrary to the reported specificity of this class of compound as inhibitors of cytochrome P-450. PMID- 8221969 TI - Studies on the constituents of Aster scaber Thunb. V. Structures of six new echinocystic acid glycosides isolated from the herb. AB - Six new echinocystic acid 3,28-O-bisdesmosides, scaberosides Hc2, Hd, Hf, Hg, Hh and Hi, were isolated as their methyl esters from the ground part of Aster scaber THUNB. (Compositae), and their structures were determined based on spectral and chemical evidence as follows. Scaberoside Hc2 is 3-O-[O-beta-D-xylopyranosyl-(1- >3)-beta-D-glucopyranosiduronic acid] echinocystic acid 28-(O-beta-D xylopyranosyl-(1-->3)-O-beta-D- xylopyranosyl-(1-->4)-O-alpha-L-rhamnopyranosyl (1-->2)-[O-alpha-L- rhamnopyransoyl-(1-->3)]-beta-D-xylopyranosyl)ester, scaberoside Hd, 3-O-[O-beta-D-xylopyranosyl-(1-->3)-beta-D-glucopyranosiduronic acid] echinocystic acid 28-(O-beta-D-xylopyranosyl-(1-->3)-O-beta-D- xylopyranosyl-(1-->3)-[O-beta-D-xylopyranosyl-(1-->4)]-O-alpha-L- rhamnopyranosyl (1-->2)-[O-alpha-L-rhamnopyranosyl-(1-->3)]-beta-D- xylopyranosyl) ester, scaberoside Hf, 3-O-[O-beta-D-galactopyranosyl-(1-->2)-beta-D glucopyranosiduronic acid] echinocystic acid 28-(O-beta-D-xylopyranosyl-(1-->4)-O alpha-L- rhamnopyranosyl-(1-->2)-[O-alpha-L-rhamnopyranosyl-(1-->3)]-beta-D- xylopyranosyl) ester, scaberoside Hg, 3-O-[O-beta-D-galactopyranosyl-(1-->2)-beta D-glucopyranosiduronic acid] echinocystic acid 28-(O-beta-D-xylopyranosyl-(1-->3) O-beta-D- xylopyranosyl-(1-->4)-O-alpha-L-rhamnopyranosyl-(1-->2)-[O-alpha-L- rhamnopyranosyl-(1-->3)]-beta-D-xylopyranosyl) ester, scaberoside Hh, 3-O-(O-beta D-galactopyranosyl-(1-->2)-[O-beta-D-xylopyranosyl-(1-->3)]- beta-D glucopyranosiduronic acid) echinocystic acid 28-(O-beta-D-xylopyranosyl-(1-->3)-O beta-D- xylopyranosyl-(1-->4)-O-alpha-L-rhamnopyranosyl-(1-->2)-[O-alpha-L- rhamnopyranosyl-(1-->3)]-beta-D-xylopyranosyl) ester and scaberoside Hi, 3-O-[O beta-D-galactopyranosyl-(1-->2)-beta-D-glucopyranosiduronic acid] echinocystic acid 28-(O-beta-D-xylopyranosyl-(1-->3)-O-beta-D-xylopyranosyl-(1-->3)-[O-bet a-D xylopyranosyl-(1-->4)]-O-alpha-L-rhamnopyranosyl-(1-->2)-[O-alpha-L- rhamnopyranosyl-(1-->3)]-beta-D-xylopyranosyl) ester. PMID- 8221970 TI - Saponin and sapogenol. L. On the constituents of the roots of Glycyrrhiza uralensis Fischer from Xinjiang, China. Chemical structures of licorice-saponin L3 and isoliquiritin apioside. AB - From the air-dried roots of Glycyrrhiza uralensis Fischer collected in Xinjiang province, China ("Shinkyo-Kanzo" in Japanese), a new oleanene-type triterpene oligoglycoside named licorice-saponin L3 and a new chalcone oligoglycoside named isoliquiritin apioside were isolated together with glycyrrhizin, 18 alpha glycyrrhizin, apioglycyrrhizin, araboglycyrrhizin, licorice-saponins A3, E2, G2, and H2, and six known flavonoid glycosides. On the basis of chemical and physicochemical evidence, the structures of licorice-saponin L3 and isoliquiritin apioside were elucidated as 3 beta-[alpha-L-rhamnopyranosyl(1-->2)-alpha-L arabinopyranosyl(1--> 2)-beta-D-glucuronopyranosyloxy]-22 beta-acetoxy-24 hydroxyolean-12-en-30-oic acid (1) and 4-O-[beta-D-apiofuranosyl(1-->2)-beta-D- glucopyranosyl]isoliquiritigenin (6), respectively. PMID- 8221971 TI - Synthesis and antiarrhythmic activity of disubstituted phenylpyridine derivative. AB - A series of disubstituted phenylpyridine derivatives was synthesized and their antiarrhythmic effects against chloroform-induced ventricular arrhythmias in mice were examined. Among them, 2- and 3-[2-(3-aminobutyramido)-4-(2,2,2 trifluroethoxy)phenyl]pyri dines (23h, 24h) and 3-[2-(3-aminobutyramido)-4 ethoxyphenyl]pyridine (24i) showed potent antiarrhythmic activity. They had approximately twice the potency of mexiletine (III). Compound 24i was selected from this series as a candidate for further development; it was found to have a class I B electrophysiological character and to show a slow kinetic rate dependent block (RDB) of the sodium channel in cardiac muscle. PMID- 8221972 TI - Synthesis of prolyl endopeptidase inhibitors and evaluation of their structure activity relationships: in vitro inhibition of prolyl endopeptidase from canine brain. AB - By chemical modification of a known prolyl endopeptidase (PEP) inhibitor (N-[N-(4 phenylbutanoyl)-L-prolyl]pyrrolidine; SUAM-1221), several arylalkanoyl derivatives (V-1--27) were synthesized and tested for in vitro inhibitory activity towards PEP from canine brain. Among them, 4-(2-thienyl)butanoyl derivatives (V-24--27) showed more potent PEP-inhibitory activity than SUAM-1221. The structure-activity relationships of these compounds are discussed. PMID- 8221973 TI - Synthesis and pharmacological properties of N-[4-[4-(1H-indol-3-yl) piperidinoalkyl]-2-thiazolyl]alkanesulfonamides as novel antiallergic agents. AB - A number of N-[4-[4-(1H-indol-3-yl)piperidinoalkyl]-2- thiazolyl]alkanesulfonamides (8--21) were synthesized and evaluated for their preventive effects on systemic anaphylaxis in guinea pigs. Structure-activity analysis revealed that methane- and ethanesulfonamide derivatives having a one to three methylene tether between the piperidine and thiazole rings exhibited potent activity but the introduction of a substituent on the indole part reduced the activity. Administration (100 mg/kg p.o.) of the four compounds 8, 9, 12, 13, together with ketotifen, oxatomide, terfenadine and azelastine as reference compounds, to mice revealed that only compound 8 caused no significant increase of the sleeping time induced by hexobarbital. In addition, compound 8 (10 mg/kg i.v.) did not change the electroencephalogram in conscious rabbits. These results led to the selection of N-[4-[4-(1H-indol-3-yl)piperidinomethyl]-2 thiazolyl]methanesulfon amide (8, FK613) for further development as a novel antiallergic agent. Clinical evaluation of FK613 is now in progress. PMID- 8221974 TI - Syntheses and biological activities of selenium analogs of alpha-rat atrial natriuretic peptide. AB - alpha-Rat atrial natriuretic peptide (7--28) (rANP (7--28)) and a series of its analogs in which half cystine residue(s) were substituted with half selenocystine residue(s) were synthesized by using the Fmoc-based solid-phase method followed by cyclization by means of dimethylsulfoxide (DMSO)-trifluoroacetic acid (TFA) oxidation. These analogs possess comparable activities in both receptor binding and cGMP accumulation in rat vascular smooth muscle cells to those of rAMP (7- 28). PMID- 8221975 TI - Platelet aggregation inhibitors in a Bhutanese medicinal plant, shug chher. AB - The 90% methanol-soluble fraction of a Bhutanese medicinal plant, Shug Chher, exhibited inhibition of platelet aggregation induced by platelet activating factor. Bioassay-directed fractionation led to the isolation of four new labdane diterpenoids, 3 alpha, 15-dihydroxy-labda-8(17), 13E-diene (5), 3 alpha-hydroxy labda-8(17), 13E-dien-15-oic acid (6), 3 alpha-hydroxy-labda-8(17), 12E, 14-trien 19-oic acid (7), and 3 alpha-acetoxyisocupressic acid (8) and four known diterpenoids, manool (1), 3 alpha-hydroxymanool (2), 3 alpha-hydroxy-12, 13E biformene (3), and isocupressic acid (4). The structures of the new compounds were determined spectroscopically. Compounds 2, 3, and 5 inhibited platelet aggregation. PMID- 8221976 TI - Constituents of rosaceous plants. I. Structure of new triterpenoids from Cowania mexicana. AB - In our search for possible anti-tumor-promoters, we carried out an investigation of the leaves and branches of Cowania mexicana D. DON (Rosaceae). Two new cucurbitane type triterpenes, 15-oxo-cucurbitacin F (3) and 15-oxo-23,24 dihydrocucurbitacin F (4), were isolated together with cucurbitacin F (1) and 23,24-dihydrocucurbitacin F (2). These triterpenes were inhibitorss of Epstein Barr virus early antigen activation induced by 12-O-tetradecanoylphorbol-13 acetate, a well-known tumor-promoter. The structures of 3 and 4 were determined from 2D-NMR spectral data and difference NOE experiments. PMID- 8221977 TI - A new drug delivery system (DDS) development using plasma-irradiated pharmaceutical aids. IV. Controlled release of theophylline from plasma irradiated double-compressed tablet composed of polycarbonate as a single wall material. AB - A controlled-release tablet can be obtained by oxygen plasma irradiation on the outer layer double-compressed tablets prepared from theophylline as a core material and polycarbonate derived from bisphenol A (PC) as a single wall material, making this possible that PC is of intramolecular bifunctionality, crosslinkable phenyl group and degradable carbonate group as an effect of plasma irradiation. It was shown that the dissolution profiles can be varied so as to cause release of theophylline at different rates, depending on the set of conditions chosen for tablet manufacturing as well as plasma operational conditions. PMID- 8221978 TI - Nematocidal activity of turmeric: synergistic action of curcuminoids. AB - A new curcuminoid, cyclocurcumin (IV), was isolated from the nematocidally active fraction of turmeric, the rhizome of Curcuma longa, together with three known curcuminoids, curcumin (I), demethoxycurcumin (II) and bisdemethoxycurcumin (III). The structure of IV was elucidated on the basis of spectral data and confirmed by the partial synthesis from curcumin (I). Although the above curcuminoids were ineffective when they were applied independently, the nematocidal activity increased remarkably when they were mixed, suggesting a synergistic action between them. PMID- 8221979 TI - Synthetic studies of vitamin D analogues. XVII. Synthesis and differentiation inducing activity of 1 alpha,24-dihydroxy-22-oxavitamin D3 analogues and their 20(R)-epimers. AB - Four vitamin D3 analogues, 1 alpha,24(S)- and 1 alpha,24(R)-dihydroxy-22 oxavitamin D3 (5 and 6) and their 20(R)-epimers (7 and 8) were synthesized from the 20(S)-alcohol (10). In tests of activity to induce differentiation of human myeloid leukemia cells (HL-60) to macrophages, 5 showed comparable activity to 1 alpha,25-dihydroxy-22-oxavitamin D3 (OCT) (2), and the other three analogues (6, 7 and 8) were less active than OCT (2). The binding properties of these analogues to the chick embryonic intestinal 1 alpha,25-dihydroxyvitamin D3 (1) receptor were evaluated. Furthermore, 20(R)-OCT (9) was synthesized and its biological properties were compared with those of OCT(2) and the 20(R)-epimers (7 and 8). PMID- 8221980 TI - Isolation and identification of anti-tumor-promoting principles from the fresh water cyanobacterium Phormidium tenue. AB - Bioassay-directed fractionation of the extract of the cyanobacterium P. tenue led to the isolation of the three classes of glycolipids, viz., monogalactosyl diacylglycerol (MGDG), digalactosyl diacylglycerol (DGDG), and sulfoquinovosyl diacylglycerol (SQDG) as anti-tumor-promoters. In comparing the anti-tumor promoting effect of the three classes of glycolipids with various acyl residues, MGDG and DGDG inhibited tumor promotion more intensely than SQDG. With respect to MGDG and DGDG, the inhibitory activity of each glycolipid possessing a single pair of acyl residues was also investigated after HPLC separation. Among the 17 tested MGDGs (1a--h) and DGDGs (2a--i), three of the DGDGs (2a, b, e) showed more potent inhibitory activity toward tumor promotion than the others. PMID- 8221981 TI - Convulsants from star anise (Illicium verum Hook.F.). PMID- 8221982 TI - [ studies of mechanism of the lesions developed in endemic fluorosis]. PMID- 8221983 TI - [Effects of low calcium on myocardial necrosis of Keshan disease by food preference]. AB - It was noticed that monotonous diet which imitated the composition of diet for the susceptible population of Keshan disease with a habit of food preference was prominently low in calcium and selenium. The plasma calcium ion content of rats kept on a monotonous diet was significantly lowered to merely half the content of the stock diet group with significant lowering of GSH-px activity. After peritoneal injection of furosemide, the plasma calcium ion contents of the monotonous diet group were further lowered significantly and signs of calcium deficiency, such as muscle tremor, spasm and convulsion might occur. If anoxic factor NaNO2 acted simultaneously, acute and severe myocardial necrosis developed. Morphologically, the myocardial necrosis was similar to that in Keshan disease. When monotonous diet was supplemented with only calcium, the growth state of the rats was significantly improved, and the degree of myocardial necrosis was significantly decreased. Supplement of calcium, selenium as well as vitamin E, seemed more effective. It is suggested that low selenium is the basic factor of endemic pathogenesis of Keshan disease, and low calcium intake by food preference among the susceptible population of Keshan disease plays an important role in the pathogenesis of myocardial necrosis in Keshan disease. PMID- 8221984 TI - [Immunohistochemical studies of islet lesions in Keshan disease patients and rats fed with grains from endemic area of the disease]. AB - Pancreas tissue collected from 77 patients died of Keshan disease as well as pancreas specimens of experimental rats having been fed with grains collected from Keshan disease endemic area were studied with immunohistochemical stainings. There were lesions found morphologically in the pancreatic islets of the patients, including atrophy, reduction of number, abnormal arrangement and degeneration of the islet cells. The serum insulin and connective peptide levels as well as the connective peptide secretory reserve in the islets of rats fed with endemic area grains were all markedly decreased. These findings suggest that the islet lesions in Keshan disease may be an important factor in aggravating the development of necrosis in the myocardium. PMID- 8221985 TI - [A morphologic study on hippocampus in endemic cretinism]. AB - A pathological examination on the hippocampus of brain was carried out on 5 cases of endemic cretinism. Computer image analysis was focused on the pyramidal and granular cells. There was a wide distribution of hypoplasia, and changes of the parameters involved in the pyramidal and granular cells were remarkable, which indicated that a development disorder existed in the hippocampus. Besides, a correlation was noticed between the weights of the thyroid gland and the brain as well as the parameters of the hippocampus. It is also noticed that changes of the thyroid gland correlated with the change in the CNS. Since lesions in the hippocampus were known to be closely related to dementia, it is considered that lesions found in the hippocampus constitute one of the morphologic bases of dementia in endemic cretinism. PMID- 8221986 TI - [Stereological studies on the brain development of fetuses in cretinism endemic region after supplement of iodized salt for 10 years]. AB - Epidemiologic reports indicated that in the cretinism endemic regions the incidence of cretinism decreased obviously after supplement of iodized salt but there still existed many cretinoid cases accompanied with mild impairment of intelligence and retardation of somatic development. This suggests that some factors other than iodine deficiency might also play an important role in the pathogenesis of cretinism. 21 brains of therapeutically aborted fetuses from cretinism endemic region after a supplement of iodized salt for ten years were studied under both light and electron microscopes. Serum T3, T4 and TSH of the fetuses and their mothers were coincidentally assayed. The brain development of fetuses from the endemic region was noticed to be still retarded as compared with those from the non-endemic region, despite that the serum hormones of the fetuses and their mothers in the endemic region showed no more significant difference from those in the non-endemic region. The retardation of the brain development could be evidenced by the increase of cellular density, decrease of the average volume of neurocytes, increase of the volume ratio between nucleus and cytoplasma per neuron, as well as decrease in number and average surface area of the mitochondria, and all of these were obtained in the cerebral cortex, hippocampus and the layer of Purkinje's cells of the cerebellum. PMID- 8221987 TI - [Morphometric analysis on the in vitro invasiveness of mouse lung adenocarcinoma variants with different metastatic potential]. AB - The in vitro invasiveness of LA1, LAD, LA5 variants of mouse lung adenocarcinoma (LA795) with different metastatic potentials were examined. Precultured heart fragment (PHF) was used as the target organ, and spheroid aggregates of LA1, LAD, LA5 cells were prepared by cultivation of cell suspension on a gyratory shaker. The invasion-complex was formed by confronting of the PHF and cell aggregate. The invasion-complex was then examined at different time intervals histologically and morphometrically. The micro-computer-aided image analysis system was employed to measure the total area (TA) of the invasion complex and the tumor tissue area (subarea, SA). The ratio of SA/TA was used in estimating the invasion ability. It was shown that the invasion of LA1, LAD and LA5 was progressive in nature and the invasive capability of these three variants were correlative with their metastatic potentials. PMID- 8221988 TI - [Preparation, identification and preliminary application of anti-desmoplakin I monoclonal antibody]. AB - Desmosomes were isolated first from fresh cow nose epidermis. Desmoplakin I (DP I) was purified from desmosome preparations by SDS-polyacrylamide gel electrophoresis and anti-DP I McAb was obtained via hybridoma technique. The tissue specificity showed that all the normal epithelia, different kinds of epithelial tumors and meningioma were positive, whereas non-epithelial tissues and their corresponding tumors gave a negative result. Positive staining showed a series of punctate reaction locating along the cell-cell borders, each spot representing a desmosome or a cluster of closely spaced desmosomes. It is suggested that McAb of DP I is a reliable immunohistochemical probe for epithelial tumor and meningioma diagnosis. PMID- 8221989 TI - [The ultrastructural pathology of chronic hepatitis C]. AB - Electron microscopy of 36 liver biopsies from patients of post-transfusion chronic hepatitis C was carried out. All of the cases were confirmed beforehand through histological examination and serological tests including identification with anti-HCV antibody. No ultrastructural changes identical with those described in experimentally induced hepatitis C were detected, though some ultrastructural alterations, not pathognomonic, did occur in the hepatocellular cytoplasm. Prominent proliferation of smooth endoplasmic reticulum was frequently found, notably in cases of chronic persistent hepatitis, and rough endoplasmic reticulum associating with distended cisternae filled with light fluffy material, occasionally forming curved membraned structure in the cisternae were also noticed. Intercellular and perisinusoidal fibrosis, as well as invasion of lymphocytes into Disse and intercellular spaces, nevertheless, without concomitant liver cell injury in the immediate vicinity were noted in the study. PMID- 8221990 TI - [Condyloma acuminatum: histopathology and detection of human papilloma virus]. AB - Comparing with the results of HPV DNA detection by PCR, 44 cases of condyloma acuminatum and condyloma-like lesions in female genital tract were studied histopathologically. Among 40 cases formerly diagnosed as condyloma acuminatum or suspicious condyloma acuminatum, all were HPV DNA detectable. HPV6 and HPV11 were found in 4 cases without koilocytes. HPV6 was found in one out of 4 cases formerly diagnosed as pseudocondyloma. The result showed that koilocytes are characteristic and diagnostic, anyhow, undiscovery of these cells still cannot rule out the possibility of condyloma acuminatum. PMID- 8221991 TI - [Smooth muscle cell migration induced by macrophage-derived chemotactic factor]. AB - The chemotactic activity of rabbit peritoneal macrophage condition-medium (M phi CM) was examined through micropore filter assay. The effects of M phi-CM on smooth muscle cell (SMCS) migration were considered to be chemotactic rather than chemokinetic in nature. This chemotactic activity was heat stable at 80 degrees C for 30 min, but was abolished at 100 degrees C for 15 min. The chemotactic activity was still maintained after dialysis, but was abolished after incubation with trypsin. It is suggested that the chemotactic factor in M phi-CM may be a protein. PMID- 8221992 TI - [The effect of hypoxic endothelial cells conditioned medium on the growth of pulmonary artery smooth muscle cells]. AB - The effect of hypoxic endothelial cells conditioned medium (HECCM) on porcine pulmonary artery smooth muscle cells (PASMC) were studied with phase contrast microscopy, 3H-thymidine labelling technique and flow cytometric analysis. The results showed that HECCM could stimulate PASMC proliferation, by promoting the cells from G0/G1 phase into S phase, and increased 3H-thymidine incorporation. Direct hypoxic condition could inhibit cultured PASMC proliferation, and arrested them at the G0/G1 phase, accompanied with decrease of 3H-thymidine incorporation. Statistical significance was obtained between data of these two groups respectively (P < 0.01). It is believed that PAEC may be the target cell to hypoxic injury, and hypoxia may lead PAEC to secrete some growth factors in auto or paracrine manner which is effective in promoting PASMC proliferation. PMID- 8221993 TI - [Ultrastructural changes in endothelial cells in rabbits with multiple organ failure (MOF)]. AB - The morphological lesions of endothelial cells in rabbits with MOF were studied by scanning (SEM) and transmission electron microscopy (TME). The edges of the cells were blurred in the MOF groups. TEM showed that the nuclei were irregular and the mitochondria were swollen. The results indicated that the vascular endothelial cells were injured in rabbits with MOF in various extent. Administration of vitamin E in the early stage seemed beneficial in protecting the endothelial cell in this event. PMID- 8221994 TI - [The morphologic changes of rat astrocytes treated with ammonium chloride in vitro]. AB - Primarily cultured astrocytes were treated with ammonium chloride in various concentrations, namely 5mM, 10mM and 15mM. The changes were does-and time dependent. After 1 to 4 days of treatment, there were mild vacuolization, eosinophilic granulation, hypertrophy and hyperplasia with increasing mitotic figures and enhanced GFAP reactivity. The number of cells and mitoses were four and five times more than those of the normal controls respectively (P < 0.01). The cells displayed an increase in number and width of the processes. The mean area, perimeter as well as the form factors were significantly increased as compared with those of the control (P < 0.01). While in the later stage (7-10 days after NH4Cl treatment), lamellation and fragmentation of the cellular processes, remarkable vacuolization and granulation as well as appearance of eosinophilic bodies in the cytoplasm were obtained accompanied with vacuolated or pyknotic nuclei. The GFAP reactivity became weak even negative. The results indicated that to the administration of ammonium chloride, the astrocytes had a dynamic plasticity. PMID- 8221995 TI - [Malignant melanoma of the sole]. AB - Ten cases of malignant melanoma on the sole are reported. Of these, 4 were acral lentiginous melanoma (ALM); 3 nodular melanoma; 2 were unclassifiable and 1 was ALM in situ. The nodular melanoma on the sole possessed a common feature with that occurring at other area of the body. ALM was considered consisting of a lump and its surrounding pigmented macular areas, while ALM in situ only appeared as a pigmented macula. Usually, the pigmented macular lesions were noticed after middle-age, and coincidentally with its gradual enlargement, its color changed from pale brown to black, while the border of the lesion was either sharp or vague with angular notches. Recognition of these features may facilitate early diagnosis and excision. Additionally, it was found that the degree of proliferation, distribution patterns and extent of atypical melanocytes in the epidermis related correspondingly to the shade variation of the pigmented maculas. PMID- 8221996 TI - [Evaluation of immunochemistry staining in melanocarcinoma diagnosis]. AB - Eleven cases of small cell tumor suspected of melanocarcinoma and another 10 cases of confirmed melanocarcinoma were studied with S-100, HMB-45, NSE and c-k antiserum staining. The main puzzling cells influencing to make a final diagnosis of melanocarcinoma by HE staining were those small cells similar to nevus cells and those round cells with abundant eosinophilic cytoplasm. Nevertheless, in both of them, there were no pigment obtained. In MF, S-100, HMB-45 and NSE staining, all of the 10 cases diagnosed as typical melanocarcinoma were positive except that one case was negative to S-100 and NSE staining. Among 11 suspicious cases, 8 were negative to MF (3 positive) and 9 cases were positive to S-100 and HMB-45 stains. The 2 negative cases left, anyhow, were positive to C-k staining which was finally confirmed as carcinoma rather than melanoma. PMID- 8221997 TI - [A high affinity CD3 monoclonal antibody HIT3a. I. Production and identification]. AB - A monoclonal antibody (McAb, named HIT3a) against CD3 antigen of human T lymphocytes was produced by the mouse-mouse hybridization technique and identified by immunofluorescent test, immunohistochemistry assay, comparison with standardized McAbs, and some studies of biological properties. HIT3a McAb was found to have high affinity, good stability, complement fixation activity, specific cytotoxicity against T cells, and effective mitogenic activity on normal PBMCs. So it might be a good reagent for clinical and laboratory research. PMID- 8221998 TI - [Effects of antihypertensive factor on cardiac function in spontaneously hypertensive rats]. AB - In this study, we observed the effects of erythrocytic AHF from normotensive human subjects on heart function of stroke prone SHR (SHRsp) in order to investigate the antihypertensive mechanism of AHF. The results showed that AHF significantly decreased the heart rate (HR), left ventricular end systolic pressure (LVESP), and magnitude of +/- LVdP/dtmax, but had no effect on Wistar Kyoto (WKY) rats, indicating that AHF has negative chronotropic and inotropic action on SHRsp hearts. AHF also lowered the contraction amplitude and resting tension in isolated SHRsp hearts, but had no effect on HR and blood flow of the coronary artery, indicating that AHF exerts negative inotropic action on isolated SHRsp hearts. The results suggest that the antihypertensive mechanism of AHF may be related to its ability to decrease cardiac function in hypertensive rats. PMID- 8221999 TI - [The influence of DPDPE on intracellular free calcium concentration induced by Bt2 cAMP and phorbol ester]. AB - The effect of DPDPE on intracellular free calcium concentration in neuroblastoma glioma hybrid cells (NG108-15) was studied with fura-2/AM fluorescence. The results were as follows: 1) The membrane permeable cAMP analogue Bt2cAMP, the adenylate cyclase activator forskolin and the protein kinase C activator TPA all induced an increase in intracellular free calcium concentration. 2) Verapamil suppressed the increase of [Ca2+]i induced by Bt2cAMP, forskolin and TPA. 3) DPDPE blocked the increase of [Ca2+]i induced by Bt2cAMP and forskolin, but not that induced by TPA. These results imply that DPDPE suppresses cyclic AMP dependent protein kinase-induced but not protein kinase C-induced intracellular free calcium. PMID- 8222000 TI - [Investigation of a Candida albicans-specific DNA fragment used for nucleic acid hybridization]. AB - A 2.5 kb fragment from Candida albicans Cli EcoRI-digested was labeled with 32P and used to probe blots of DNA from various organisms. DNA from all strains of C. albicans gave evidence of strong sequence homology with the 2.5 kb fragment. With the exception of weak hybridization signals from strains of C. parapsilosis and a strain of C. tropicalis, DNA from other Candida species, Cryptococcus as well as bacteria did not hybridize with the probe. The results indicate that the 2.5 kb fragment might contain sequence unique to C. albicans. After this is confirmed by cloning, a specific probe for the identification of C. albicans will be made. PMID- 8222001 TI - [Alterations of protein kinase C activity in rat myocardium and aorta smooth muscle during endotoxemia]. AB - A decline of cytosol protein kinase C activity was observed in rat myocardial cells at 4 and 8 h after endotoxin administration, and membrane-associated protein kinase C activity rose at the same time. The activity of membrane protein kinase C in aortic smooth muscle cells at 0.5 and 4 h after endotoxin injection was higher than that in control, while cytosol protein kinase C activity was lower. The results indicate that protein kinase C was activated in myocardial cells and aortic smooth muscle cells during various phases of endotoxemia. PMID- 8222002 TI - [Ultrastructural study on the epithelium of ligated oviducts in reproductive women]. AB - The epithelium of normal and ligated oviducts in nine reproductive women was studied by scanning electron microscopy. The results showed. The cilia of the epithelium were twisted and adhered on to the mucosa 0.5 cm from both the distal and proximal ends of the ligation scar. In the localized mucosa, the epithelial cells were irregular, and numerous microvilli and cilia were lost. However the microvilli and cilia of the epithelium were normal on mucosa 1.0 cm from the blocked oviducts. In the present experiment, oviductal stomata were reported on the epithelium of the hydrosalpinx for the first time. These oviductal stomata, located among some secretory cells, might be connected with the lymphatic capillaries of the oviducts. On the mucosa of the hydrosalpinx, numerous cilia were lost, and microvilli were abundant on the surface of the secretory cells. It might be possible that some secretory cells on the mucosa of the hydrosalpinx are evolved from ciliated cells. In addition, the pathogeny of the hydrosalpinx is discussed. This study shows: A) The transportation of the ovum is compromised due to damage to the cilia and microvilli of the oviductal epithelium, and this might be an important cause of continued infertility following reversal of sterilization; B) It is necessary to remove mucosa at least 0.5 cm from both the distal and proximal tubal segments of the ligation scar during the operation of end-to-end anastomosis; C) The oviductal stomata may be involved in draining hydrops from the hydrosalpinx. PMID- 8222003 TI - [Effect of hyperglycemia on insulin release from isolated rat pancreatic islets]. AB - We investigated the long-term (24h) influence of high concentration glucose (26.4mmol/L) on insulin release by culturing collagenase isolated rat pancreatic islets in vitro. The cultured islets were morphologically normal by histological examination. Compared with freshly isolated islets, islets cultured in 26.4mmol/L glucose showed higher basal insulin release (P < 0.05). The effect of high glucose was time dependence. When islets preexposed to high glucose medium for 24h were perfused, the basal insulin release was significantly higher than that of freshly isolated islets, the first-phase glucose stimulated insulin release disappeared, and the second-phase was diminished. Islets cultured in high glucose medium showed increased sensitivity to L-arginine. Long-term high concentration glucose could potentiate the arginine-induced release. When added to the culture medium with 26.4mmol/L glucose, Ginseng saponins markedly increased the responsiveness of islets to glucose stimulation, and augmented the sensitivity of incubated islets to additional Ginseng saponins. PMID- 8222004 TI - [The suppressive effect of Tripterygium wilfordii hook F on the IL-2 autocrine loop of human T cells]. AB - We studied the effect of Tripterygium wilfordii Hook F on the IL-2 autocrine loop of human T cells in vitro. The results revealed that GTW significantly inhibited the activation of T cells production of IL-2, expression of IL-2R and proliferation response of activated T cells exogenous IL-2. PMID- 8222005 TI - [The turnover of three amino acids in the plasma of chronic renal failure patients]. AB - In this study, three amino acids (Ala, Leu and Phe) labelled with stable isotopes were administered to a group of CRF patients and of normal controls, respectively. Based on the measurement of gas chromatography and mass spectrometry, a series of kinetic parameters of amino acid metabolism were calculated and the differences between the two groups were compared. PMID- 8222006 TI - [The effect of IH764-3 on proliferation and function of fibroblasts]. AB - IH764-3, a potent component isolated from Salviae miltiorrhizae (a component of TML) was used to study the effect on proliferation and functions of cultured fibroblasts. The fibroblast growth curve demonstrated a dose-dependent relationship between growth and IH764-3 concentration. The incorporation of 3H TdR and 3H-proline into fibroblasts was significantly inhibited by IH764-3. Calmodulin level, fibronectin and thrombospondin contents in the test group were obviously lower than those in the control group. Flow-cytometry showed that in the IH764-3 treated group, the percentage of cells in G0 + G1 phase was higher than that in the control. Electron microscopic observation (TEM and SEM) showed that in the treated group, the secretory function of collagen had decreased. All the results indicated that IH764-3 exerts a direct inhibitory effect on fibroblast proliferation and affects their ability to synthesize and secrete collagenous substances. PMID- 8222007 TI - [Measurement of human erythrocyte membrane potential with microelectrode]. AB - The measurement of erythrocyte membrane potential (Ery-MP) in 189 healthy subjects using a microelectrode showed that the Ery-MP in 96 healthy subjects was -5.80 +/- 0.31 mV, and that in 93 healthy female subjects was -6.30 +/- 0.34 mV. Plasma lipid peroxide (P-LPO), erythrocyte lipid peroxide (Ery-LPO), plasma superoxide dismutase activity (P-SODA) and erythrocyte superoxide dismutase activity (Ery-SODA) were measured at the same time. After linear regression and correlation analysis, it was found that the Ery-MP value of healthy adults decreased as the P-LPO and Ery-LPO values increased, displaying a linear negative correlation, Ery-MP and Ery-SODA values formed a linear positive correlation. PMID- 8222008 TI - [Cytotoxicity to hepatoma BEL-7402 cells of an antitumor monoclonal antibody streptonigrin conjugate]. AB - Streptonigrin, a highly active antitumor antibiotic, was covalently conjugated to anti-human hepatoma monoclonal antibody 3A5 via the active ester method. The conjugate showed biological activities and UV spectra characteristics of streptonigrin and McAb 3A5. The molar ratio of streptonigrin to 3A5 was 2-6:1, with protein recovery of 76%. The conjugate retained 12.5% of drug activity and nearly full antibody activity, though as the number of streptonigrin molecules in the conjugate increased, the antibody activity of the conjugate decreased. As determined by clonogenic assay with human hepatoma BEL-7402 cells in vitro, the inhibitory potency of the conjugate was 63-fold stronger than that of free streptonigrin. For KB cells (which react weakly with 3A5) the cytotoxicity was 11 fold weaker. The results indicate that the McAb 3A5-streptonigrin conjugate is selective for target cells. PMID- 8222009 TI - [High-resolution cytogenetic study of a patient with Prader-Willi syndrome]. AB - We investigated a patient with Prader-Willi syndrome by using the high-resolution banding technique. His clinical findings were also examined in some detail. The patient had interstitial deletion of 15q, with breakpoints at 15q11.2 and 15q13. However, he had a normal karyotype at about 400 bands per haploid-set. During the course of this study, it was realized that a small deletion in the proximal 15q could be easily overlooked when a mitotic spread of about 400-bands or less per haploid-set was used. It is suggested that the high-resolution banding technique is of great importance in identifying microdeletions and other microaberrations of chromosomes. PMID- 8222010 TI - [Bronchial disruption due to blunt chest trauma--a report of 6 cases]. AB - Six cases with bronchial disruption due to blunt chest trauma are reported. Clinical symptoms were dyspnea in 5 patients, hemopneumothorax in 3 cases, and hemoptysis and subcutaneous emphysema in 4 cases each. Pneumothorax was not found on immediate radiological examination in 3 patients. Bronchial disruption was defined by exploratory thoracotomy in one case three days after trauma. In the remaining 5 cases, the diagnosis was demonstrated by lipiodol bronchography and fibrobronchoscopy 70 days to 6 years later. End-to-end anastomosis of the bronchus was performed in 5 patients, whereas right middle and lower lobectomy was done in one case. Five patients with bronchial repair suffered transient atelectasis of the lung, which reexpanded in 4 to 10 days. PMID- 8222011 TI - [The structure of bovine choriocapillaris]. AB - The ultrastructure of bovine choriocapillaris was investigated in bovine choroid and isolated choriocapillary samples. The choriocapillaris lies in the layer just outside the Bruch's membrane. The choriocapillary endothelial cells on the side facing Bruch's membrane were found tapering their cytoplasm, with a significant number of fenestrae. The fenestrae were predominantly located in the choriocapillaris at the posterior pole, rather than at the periphery, of the eyeball. The endothelial cells on the side facing the sclera had few fenestrae, and the nuclei of endothelial cells and pericytes were located on this side. It was found that on the peripheral part of the choroid, the basement membranes of either choriocapillary endothelial cells or those of pericytes take part in composing the outer part of the Bruch's membrane. The isolated choriocapillaris revealed a typical lobular pattern of capillaries arranged in a radiating fashion. Intercellular junctions and/or cell contacts among endothelial cells and pericytes were also investigated. PMID- 8222012 TI - [The veterinarian yesterday and tomorrow]. PMID- 8222013 TI - [Fetal and adrenal transplantation in Parkinson disease: theoretical bases and clinical results]. PMID- 8222014 TI - [Heat shock proteins]. AB - All cells, procaryotic and eucaryotic, respond to a rise in environmental temperature by a rapid synthesis of a set of specific proteins: the heat shock proteins (HSPs). These HSPs appear to be among the most conserved proteins in nature and certain members of the HSP family are present in non-stressed cells. These observations are reflecting the fact that HSPs serve vital physiological functions as well in normal cells as in stressed cells. In the past few years, various fields of medicine have shown growing interest in the role that the HSPs may play in cellular mechanisms. In this paper we are briefly summarizing the actual knowledge of HSP involvement in carcinogenic and immunological processes. PMID- 8222015 TI - [The first 7 months of activities in a psychogeriatric unit in Luxembourg]. AB - Since September 1991 a new unity has been opened in the Neuro-Psychiatric Hospital: a ward of 26 beds for psychogeriatric patients. 50 persons (half women and half men) have been admitted during the first seven months. By the end of this study (June 1992) 76% are no more staying in the hospital. Besides the five deaths, most of the people have returned to their previous home. With a mean stay of two to three months we believe that our psychogeriatric ward will be able to admit about 70 or 80 new patients by one year. PMID- 8222016 TI - [Chemoprevention: a new strategy in the fight against breast cancer?]. PMID- 8222017 TI - [Theophylline and seizures]. PMID- 8222018 TI - Doctors' attitudes towards patients who smoke and opinions on the role of the medical profession in policies to control tobacco use. PMID- 8222019 TI - An unusual presentation of latex allergy. PMID- 8222020 TI - Unilateral pneumothorax--an unexpected complication of laparoscopic cholecystectomy. PMID- 8222021 TI - Cervical oro-pharyngeal oedema and severe hypoacusia: complication of antecubital vein catheterization. PMID- 8222022 TI - "Pre-emptive analgesia" may follow epidural anaesthesia. PMID- 8222023 TI - Anaesthesia for awake craniotomy. PMID- 8222024 TI - Endotracheal tube fires. PMID- 8222025 TI - Combined loss of resistance-free flow hanging drop technique for spinal anaesthesia. PMID- 8222026 TI - Vaporizer overfilling. PMID- 8222027 TI - Anaesthesia oral examination. PMID- 8222028 TI - Adverse events in anaesthesia: the role of equipment. PMID- 8222029 TI - Psychoactive substance use among American anesthesiologists: a 30-year retrospective study. AB - The purpose of this study was to assess the cumulative incidence of substance use among anesthesiologists during training and practice, the effect of stress on drug use, and deterrent efficacy of institutional prevention programmes. The 260 anesthesiologists who had trained at the Medical College of Wisconsin between 1958-1988 were surveyed by mail regarding psychoactive substance use. Analysis of 183 responses focused on demographic and psychosocial factors. Substances used most frequently included: alcohol (91.6%), marijuana (30.8%) and cocaine (9.4%). Twenty-nine (15.8%) anesthesiologists were identified as being substance dependent: 19 were alcohol-impaired; six were drug-impaired, and four were dependent on both alcohol and drugs. Impairment was more prevalent in anesthesiologists who had completed their training after 1975. Fifty-eight (32%) anesthesiologists had used illicit drugs to "get high"; 11 acknowledged daily use for two weeks or more, with eight admitting dependency. Substance abuse was more common in parents of impaired anesthesiologists (35.7%) than in unimpaired colleagues (8.1%; P < 0.001). The divorce rate for impaired anesthesiologists (24.1%) was greater than for unimpaired anesthesiologists (5.2%; P < 0.001). Increased stress during training was not reflected by increased substance use. Few recalled any drug counseling whatsoever. Seventy percent assessed hospital drug control policies as fair or poor. Younger respondents (born after 1951) were more critical of drug control programmes than their older cohort. Incidents of substance abuse were reported for both residents and faculty. Psychoactive substance abuse remains a serious problem among anesthesiologists. PMID- 8222030 TI - Recall of intraoperative events after general anaesthesia and cardiopulmonary bypass. AB - We wished to identify patients able to recall intraoperative events after general anaesthesia involving cardiopulmonary bypass (CPB). A balanced anaesthetic technique consisting of benzodiazepines, low dose fentanyl (15.9 +/- 8.5 micrograms.kg-1) and a volatile agent was employed. Perioperative recall was sought utilizing a structured interview on the fourth or fifth postoperative day. During 20 mo 837 patients underwent CPB. Seven hundred patients (84%) were able to respond to a structured postoperative interview. A detailed chart review was performed in patients with recall and in 60 randomly selected patients without recall. Eight patients (1.14%) reported recall of intraoperative events. We were unable to identify any differences between the two groups with respect to narcotic, benzodiazepine dosage or usage of inhalational agents. The incidence of recall in patients undergoing cardiac surgery was less in our group than previously reported. It is, however, higher than the 0.2% incidence recently reported in patients undergoing non-cardiac surgery. This is probably due to patient characteristics and intraoperative factors which make it difficult to avoid periods of relatively light anaesthesia during cardiac surgery. PMID- 8222031 TI - Propofol anaesthesia reduces early postoperative emesis after paediatric strabismus surgery. AB - Propofol anaesthesia may reduce postoperative emesis. The purpose of this study was to compare the incidence of emesis after propofol anaesthesia with and without nitrous oxide, compared with thiopentone and halothane anaesthesia, in hospital and up to 24 hr postoperatively, in outpatient paediatric patients after strabismus surgery. Seventy-five ASA class I or II, unpremedicated patients, aged 2-12 yr were randomly assigned to one of three groups: Thiopentone, 6.0 mg.kg-1 i.v. induction followed by halothane and N2O/O2 for maintenance (T/H); propofol for induction, followed by propofol and oxygen for maintenance (P/O2); and propofol for i.v. induction, followed by propofol infusion and N2O/O2 for maintenance (P/N2O). All received vecuronium, controlled ventilation, and acetaminophen pr. Morphine was given as needed for postoperative analgesia. There were no differences in age, weight, number of eye muscles operated upon, duration of anaesthesia or surgery. The P/N2O group (255 +/- 80 micrograms.kg-1 x min-1) received less propofol than the P/O2 group (344 +/- 60 micrograms.kg-1 x min-1) (P < or = 0.0001) and had shorter extubation (P < 0.001) and recovery (P < 0.01) times. Emesis in the hospital, in both the P/N2O (4.0%) and P/O2 group (4.0%) was less than in the T/H group (32%) (P < 0.01). Antiemetics were required in four patients in the T/H group (16.0%). Overall emesis after surgery was not different among the groups: T/H (48%), P/O2 (28%) and P/N2O (42%). The use of propofol anaesthesia with and without N2O decreased only early emesis. This supports the concept of a short-acting, specific antiemetic effect of propofol. PMID- 8222032 TI - Oral midazolam premedication for children with congenital cyanotic heart disease undergoing cardiac surgery: a comparative study. AB - To determine whether oral midazolam is a safe and effective alternative to our current standard premedication for children with cyanotic congenital heart disease (CCHD), 30 children aged 1-6 yr, scheduled for elective cardiac surgery, were studied. The children were randomly assigned to one of two groups: Group I received oral midazolam 0.75 mg.kg-1 30 min before separation from their parents in the surgical waiting area, and Group II received oral or rectal pentobarbitone 2 mg.kg-1 at 90 min, and morphine 0.2 mg.kg-1 and atropine 0.02 mg.kg-1 im at 60 min before separation. Heart rate, haemoglobin oxygen saturation (SpO2) and anxiolysis and sedation scores were recorded at four times during the study: at baseline (immediately before premedication), immediately after administration of the premedication, at separation of children from parents in the waiting area and at the time of application of the face mask in the operating room. We found that in Group I, anxiolysis improved at separation from parents compared with baseline (P < 0.05) and sedation increased both at separation and on mask application (P < 0.05), whereas in Group II anxiolysis did not change at any time and sedation increased only at separation (P < 0.05). Intramuscular injection of morphine produced a transient decrease in mean SpO2 (from 84% to 76%) (P < 0.05) that did not occur after ingestion of oral midazolam. The results of this study indicate that oral midazolam is a safe and effective replacement for the standard premedication for children with CCHD undergoing cardiac surgery and avoids the decrease in SpO2 associated with im injections. PMID- 8222033 TI - Prolonged duration of succinylcholine in patients receiving anticonvulsants: evidence for mild up-regulation of acetylcholine receptors? AB - Succinylcholine (SCh) normally causes a small increase in serum potassium concentration, but certain conditions may predispose to severe hyperkalaemia. This is due to "up-regulation" of skeletal muscle acetylcholine receptors (AChR), which also results in resistance to non-depolarizing muscle relaxants (NDMR). Anticonvulsant therapy causes NDMR resistance because of sub-clinical blockade, and diminished release, of acetylcholine. We studied nine patients chronically receiving anticonvulsants (phenytoin and/or carbamazepine) and nine control patients. Anaesthesia was induced typically with thiopentone or propofol; isoflurane and N2O were used for maintenance. The ulnar nerve was supramaximally stimulated and mechanical twitch height was measured with a force transducer at the adductor pollicis, before and after SCh 1 mg.kg-1, until return to baseline height. Plasma potassium concentration was measured before and at three, five, and ten minutes following SCh. Mean maximum potassium rise was 0.2 mEq.L-1 in each group. The time for return to baseline twitch height was 14.3 +/- 2.3 min (mean +/- SD) in the anticonvulsant group and 10.0 +/- 1.6 min in the control group, P = 0.001. The recovery index (time for 25% to 75% recovery) was 2.6 +/- 0.9 min in the anticonvulsant group and 1.4 +/- 0.3 min in the control group, P < 0.01. The normal potassium response coupled with prolonged duration suggests a hypersensitivity to SCh that is consistent with an anticonvulsant-induced mild up regulation of AChR. PMID- 8222034 TI - Posture and the spread of hyperbaric bupivacaine in parturients using the combined spinal epidural technique. AB - This study was undertaken to analyze the effect of posture on the spread of hyperbaric bupivacaine in pregnant women using a combined spinal extradural technique, and to assess the quality of analgesia provided by 10 mg bupivacaine when using this technique. Fifty parturients undergoing elective Caesarean section under regional anaesthesia were randomly allocated to receive 2.0 ml hyperbaric bupivacaine 0.5% in either the sitting or left lateral position. Spinal injection was performed with a 27 gauge, 120 mm long spinal needle using a single space combined spinal extradural technique. The onset time to analgesia at T4, and grade 3 motor block was on average 7.7 min and 6.9 min respectively in the lateral group, compared with 10.8 min (P < 0.05) and 9.4 min (P < 0.05) in the sitting group. Nine women in the sitting group and one woman (P < 0.05) in the lateral group required epidural supplementation. Hypotension occurred in 48% of the parturients in the lateral group and in 13% (P < 0.05) of the parturients in the sitting group. Nausea was noted in 61% of the parturients in the lateral group and in 22% (P < 0.05) of the parturients in the sitting group. There was no difference between the two groups in neonatal outcome. Overall, the position of the patient during induction of spinal anaesthesia does influence the rate of onset of analgesia and motor blockade. Injection of 10 mg hyperbaric bupivacaine in the sitting position would not provide adequate analgesia for Caesarean section when using a single space combined spinal extradural technique. PMID- 8222035 TI - The oral examination in anaesthetic resident evaluation. AB - Oral examinations have a traditional place in training and evaluation of professionals. Despite a lack of evidence identifying their value in assessment of candidates seeking anaesthetic specialty certification, oral examinations continue to be widely used. Although there is a considerable body of literature concerning oral examinations, there is no description of how this technique is employed in anaesthesia in Canada. The objective of this review is to provide faculty and residents with information concerning the strengths and weaknesses of this format, and the structure of the oral examination as practised in anaesthesia. Reliability of oral examinations can be affected by a number of factors dependent on the examiner, candidate, and the format. Properly constructed and prepared questions have well-defined characteristics. Components tested during the oral examination include: evaluation of a clinical situation, choice of therapy, medical knowledge, ability to deal with emergency situations, decision-making ability, and communication skills. When appropriately planned, the oral examination can be a useful component of the certification process. PMID- 8222036 TI - Coeliac plexus block for chronic pain syndromes. AB - The purpose of this article is to familiarize the anaesthetist with the basic anatomy of the coeliac plexus; the techniques used to perform the procedure, its indications, complications and results in the management of chronic abdominal pain syndromes. Radiological, surgical and anaesthetic literature from the beginning of the century were reviewed. The main indication for neurolytic coeliac plexus block is intractable pain secondary to carcinoma of the pancreas or stomach. There appear to be theoretical advantages to techniques that result in spread of solution anterior to the aorta, such as the trans-aortic approach. These have not yet been demonstrated in any studies with large numbers of patients. PMID- 8222037 TI - Severe pulmonary oedema after venous air embolism. AB - We present a 59-yr-old Chinese male patient who developed acute pulmonary oedema and cardiovascular collapse following multiple episodes of venous air emboli while in the sitting position for removal of a cervical meningioma. The severity of the pulmonary oedema and cardiovascular disturbance were surprising. Postoperative ventilation and inotropic support were required and five litres of plasma were needed to replace the fluid lost as pulmonary oedema. We discuss the differential diagnosis of the pulmonary changes and review current ideas on the pathogenesis for pulmonary oedema following venous air embolism. PMID- 8222038 TI - Severe respiratory depression after epidural morphine in a patient with myotonic dystrophy. AB - We describe a patient with myotonic dystrophy who underwent cholecystectomy, and developed severe respiratory depression following epidural administration of morphine to provide postoperative analgesia. At preoperative assessment, he demonstrated near normal vital capacity and maximal voluntary ventilation, but the presence of chronic ventilatory failure with a resting value of PaCO2 51 mmHg. Anaesthesia was produced by a combination of epidural and light general anaesthesia without intravenous anaesthetics, narcotics or neuromuscular relaxants. Five hours after epidural administration of 2 mg morphine, the patient developed severe respiratory depression with a PaCO2 of 93 mmHg. Intravenous naloxone resulted in transient improvement in minute volume, suggesting that epidural morphine was responsible for the depression. Epidural morphine can cause unexpected respiratory depression, even at a small dose, because of the sensitivity of the respiratory centre to morphine in patients with myotonic dystrophy. PMID- 8222039 TI - Amniotic fluid embolus. AB - A case report of a 27-yr-old healthy patient for Caesarean section under epidural anaesthesia is presented. The patient suffered an acute cardiorespiratory collapse when the infant's head was being delivered through the anterior abdominal wall. The patient remained cyanosed after proper tracheal intubation and pulmonary ventilation with 100% O2. Hypotension was difficult to treat but returned to normal 25 min after the event. A pulmonary artery catheter inserted three hours after the event showed normal pressures and a high cardiac output. The patient suffered permanent neurological damage. The differential diagnosis is discussed and current concepts of the aetiology and management of amniotic fluid embolism reviewed. PMID- 8222040 TI - Anaesthesia for non-cardiac surgery in heart-transplanted patients. AB - This review documents the anaesthetic management, haemodynamic function and outcome in 18 of 86 heart-transplanted recipients, who returned for 32 non cardiac surgical procedures at the Toronto Hospital from 1985 to 1990. General anaesthesia was administered in eight of the 27 elective operations and four of the five emergency operations. Induction medications included thiopentone (2-4 mg.kg-1), fentanyl (1-7 micrograms.kg-1) and succinylcholine (1-1.5 mg.kg-1). Anaesthesia was maintained with a combination of oxygen/nitrous oxide and isoflurane or enflurane. Muscle relaxation was maintained with vecuronium or pancuronium. No delayed awakening or unplanned postoperative ventilation was observed. Neurolept-anaesthesia was administered to 63.0% and 20.0% of the elective and emergency operations, respectively. The anaesthetics included fentanyl (25-100 micrograms) and midazolam (0.5-1.5 mg) or diazemuls (2.5-5.0 mg). Spinal anaesthesia (75 mg lidocaine) was administered to only two of the 27 elective operations. No important haemodynamic changes were observed in any anaesthetic group, but lower systolic BP was found after induction and during maintenance periods in the patients who received general anaesthesia than in those who received neurolept-anaesthesia. However, no anaesthesia-related morbidity or mortality was noted. This suggests that general, neurolept- and spinal anaesthesia do not affect haemodynamic function or postoperative outcome in heart-transplanted recipients undergoing subsequent non-cardiac surgery. PMID- 8222041 TI - Anaesthesia equipment safety in Canada: the role of government regulation. AB - Medical device problems as detected by the Health Protection Branch (HPB) of Health and Welfare Canada were studied in order to examine the frequency and safety priority of problems in the subgroup of anaesthesia devices. Data pertaining to the time period from April 1, 1987 to November 30, 1992 were gathered from the Medical Devices Notification Database and the DAFFY Reporting System. The Medical Devices Notification Database contains all notifications for newly marketed medical devices in Canada and the DAFFY Reporting System keeps record of all submitted problem reports and manufacturer recalls along with their designated safety priority status. When an important safety hazard is associated with a medical device as determined from information submitted to HPB, an Alert Medical Devices may be issued to inform hospitals and health care professionals of the problem. During the studied time period, the percentages of problem reports and recalls as well as Alerts were compared with the percentage of newly marketed anaesthesia devices. It was found that although only 2.3% of the newly marketed medical devices were classified as anaesthesia devices, 8.6% of all problem reports/recalls and 37.5% of all Alerts originated from the field of anaesthesia devices. Comparison of the percentages of problem reports/recalls (8.6%) and Alerts (37.5%) with the percentage of marketed devices for anaesthesia (2.3%) showed differences (P < 0.05). As well, the percentage of class I priority (the highest safety priority) problem reports/recalls for anaesthesia devices was 10.2% compared with 4.9% for non-anaesthesia devices (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222042 TI - Anaesthesia services and the education of anaesthetists in Nepal: a model for sustainable development? AB - In 1985 the University of Calgary in Canada and Tribhuvan University in Kathmandu, Nepal Jointly established the Diploma in Anaesthesiology (DA) programme in Nepal. To evaluate the impact of the DA Programme and provide a data base for long-term planning we conducted a national survey in 1992. We sought to describe anaesthesia manpower and workloads, and to make an inventory of facilities, equipment, and supplies in different sized hospitals. Twenty-seven hospitals providing surgical services were included, nine inside and 18 outside the Kathmandu valley. Seventeen of the 21 respondent hospitals had at least one specialist anaesthetist. The results identify both strengths and weaknesses in Nepal's anaesthesia services and provide important guidelines for planning. When the DA course was launched there were only seven specialist anaesthetists in Nepal. The shortage of anaesthetists was an important factor limiting surgical services, and after DA graduates were posted to zonal (50 bed) and regional (150 200 bed) hospitals the surgical case loads doubled. There are now about 40 specialist anaesthetists in the country, of which half are DA graduates, but many hospitals have only one anaesthetist. That isolation, plus lack of continuing education (CME), are important factors threatening quality of care. Recognizing the singular role of the DA programme in alleviating Nepal's shortage of anaesthetists, we conclude that it should be renewed and strengthened to meet the needs of the next decade. Techniques commonly used at the zonal level: regional, draw-over, and total IV anaesthesia, should be stressed. At the same time fresh initiatives are required in CME and higher education for the renewal of teaching staff.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222043 TI - Tissue and cell specific methylation, repair and synthesis of DNA in the upper gastrointestinal tract of Wistar rats treated with single doses of N-methyl-N' nitro-N-nitrosoguanidine. AB - Several potential cancer risk factors have been monitored concurrently in the upper gastrointestinal tract of young adult male Wistar rats given single (i.g.) doses of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) which readily induces forestomach tumours under these conditions. Radioimmunoassay was used to determine the formation of O6-methyl-2'-deoxyguanosine (O6-MedG) in DNA after MNNG doses of 1, 5, 25 or 50 mg/kg and was found to be highest in the pylorus, with progressively lower levels in the corpus, forestomach, duodenum, oesophagus and jejunum. Immunohistochemical procedures showed that cells with nuclei containing O6-MedG were heterogeneously distributed in these tissues. O6 Alkylguanine-DNA alkyltransferase activity in untreated animals was highest in the mucosae of the corpus, lower and relatively similar in that of the pylorus, duodenum and jejunum and lowest in the tissues of oesophagus and forestomach. Estimates of DNA synthesis and cell proliferation indicated a 5-fold increase in the DNA labelling index in the forestomach whereas perturbations of DNA synthetic activity in the other tissues of the upper gastrointestinal tract were much less marked. As a result of these changes, cells with nuclei that contained O6-MedG and were also undergoing DNA synthesis (determined by sequential immunohistochemical analysis and autoradiography) were found most commonly in the forestomach and to a lesser extent in the pylorus. This distribution of replicating damaged cells corresponds with the relative tumour yields in these upper gastrointestinal tract tissues and such cells are the probable targets in this single dose carcinogenesis regime. Thus, whilst the highest concentration of O6-MedG did not correlate tumour incidence, the overall risk for tumour induction did correlate with a significant level of DNA damage, a lower capacity for DNA repair and a marked increase in DNA synthesis over the constitutive level in the target cells. Carcinogenic risk in this system is therefore more readily determined by studying several risk factors simultaneously. PMID- 8222044 TI - Tissue and cell specific methylation, repair and synthesis of DNA in the upper gastrointestinal tract of Wistar rats treated with N-methyl-N'-nitro-N nitrosoguanidine via the drinking water. AB - Several potential cancer risk factors have been monitored concurrently in the upper gastrointestinal tract of young male Wistar rats given N-methyl-N'-nitro-N nitrosoguanidine (MNNG) via the drinking water, a regimen that induces a high yield of tumours in the pylorus and to a lesser extent in the duodenum. Radioimmunoassay was used to determine the amounts of O6-methyl-2'-deoxyguanosine (O6-MedG) formed in the tissue DNA of rats given MNNG at doses of 40 or 80 micrograms/ml for periods of 3, 6 and 12 weeks. The highest adduct concentration was found in the pylorus with progressively lower concentrations in the corpus and duodenum, jejunum, forestomach and oesophagus. Between 3 and 12 weeks these adduct levels decreased in all tissues and there was no evidence of a dose dependent accumulation of O6-MedG. When analysed by immunohistochemistry the distribution of cells with nuclei containing O6-MedG was seen to be heterogeneous in the various tissues. O6-Alkylguanine-DNA alkyltransferase activity increased during the 12 weeks of MNNG treatment in oesophagus and forestomach, but decreased to approximately 50% of the initial value in the corpus, pylorus, duodenum and jejunum. The major changes in DNA synthesis and cell proliferation were the marked upward expansion (i.e. towards the lumen) of the zone of replicating cells in the glands of the pylorus and the greatly increased numbers of replicating damaged cells (i.e. cells that contained O6-MedG whilst undergoing DNA synthesis) as determined by sequential immunohistochemical analysis and autoradiography. Such cells are the probable target cells in this chronic dose carcinogenesis regime. Although similar changes also occurred in the glands of the corpus these were of lesser extent and the changes of labelling index in the oesophagus and forestomach were relatively minor. In the duodenum, MNNG treatment led to erosion of the upper part of the glands so that the zone of cells containing O6-MedG overlapped with the zone of proliferating cells resulting in the formation of many replicating damaged cells. Thus, as in the single dose study (see preceding paper) the distribution of replicating damaged cells coincides with the tumour yield in the tissues of the upper gastrointestinal tract. As in the case of single doses of MNNG the risk factors for carcinogenesis are, a significant level of DNA damage, a lower capacity for DNA repair and an increased DNA synthetic activity, again suggesting that carcinogenic risk cannot readily be determined by studying risk factors individually. PMID- 8222045 TI - CYP1A1 mRNA levels as a human exposure biomarker: use of quantitative polymerase chain reaction to measure CYP1A1 expression in human peripheral blood lymphocytes. AB - Accurate human risk assessment requires sensitive methods to evaluate dose response relationships, especially following low level exposures. We have developed a reverse transcriptase polymerase chain reaction (RT-PCR) method to quantitative cytochrome P450-1A1 (CYP1A1) mRNA levels in human blood lymphocytes. Many polycyclic aromatic hydrocarbons (PAH) such as benzo[a]pyrene, and chlorinated PAH such as polychlorinated dibenzodioxins, dibenzofurans and biphenyls induce CYP1A1 expression through activation of an endogenous protein, the Ah receptor. Using a quantitative competitive RT-PCR method that included a synthetic internal standard we determined copy numbers of CYP1A1 mRNA in resting as well as mitogen-stimulated human blood lymphocytes. In mitogen-stimulated human blood lymphocytes assay variation was approximately 10% for measurement of this low expression gene and mRNA levels correlated well with ethoxyresorufin-O deethylase (EROD) activity. The expression of mRNA was induced 20-fold upon culturing human lymphocytes with 10 nM TCDD. In nonstimulated, uninduced lymphocytes CYP1A1 levels are extremely low (1000 copies mRNA/10(4) cells) and cannot be measured by EROD activity. Studies of CYP1A1 mRNA expression in chemically-exposed populations are in progress. PMID- 8222046 TI - DNA adducts, mutations and cancer. AB - Anyone having lectured on DNA adducts is likely to be familiar with the first question after the presentation: 'What is the relationship of DNA adducts and cancer?'. Although there are a number of reviews on the topic (1-5), my personal response, instead of going into lengthy circumstantial argumentation and hand waving, has been a quotation from Mortimer Mendelson: 'I wouldn't like to have my DNA messed up'. In this commentary I will indulge in this still circumstantial argumentation. The reason being that never before has it appeared to be so easy, or less difficult, to entertain the question. Hopefully this effort will spare colleagues confronting the same question some time. At the same time I will take the opportunity to describe developments in the identification of DNA adducts. A vivid history of the development of ideas on chemical causes of cancer has been published by Lawley. PMID- 8222047 TI - Mouse endometrium stromal cells express a polycyclic aromatic hydrocarbon inducible cytochrome P450 that closely resembles the novel P450 in mouse embryo fibroblasts (P450EF). AB - Stromal cells from mouse endometrium, E041 cells, at passages 21, 23 and 26 were metabolically active towards 7,12-dimethylbenz[a]anthracene (DMBA). The total DMBA-metabolizing activity of E041 cells was preferentially increased by benz[a]anthracene (BA) relative to 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD) treatment (7-fold by BA and 4-fold by TCDD), but the relative proportions of DMBA metabolites formed remained unchanged. Profiles of DMBA metabolites generated from E041 cell microsomes were very different from that of mouse P4501A1 but exhibited similarities to that of P450EF, the polycyclic aromatic hydrocarbon (PAH)-inducible cytochrome P450 in the mouse embryo fibroblast cell line C3H10T1/2 (10T1/2). Notably, both induced and uninduced E041 cell microsomes were very effective in the formation of the proximate carcinogen DMBA-3,4-dihydrodiol (15-20% of total) and DMBA-10,11-dihydrodiol (13-18% of total) but ineffective in forming the 7-hydroxymethyl derivative of DMBA (< 1% of total). Antibodies to P450EF completely inhibited the DMBA-metabolizing activities of both induced and uninduced E041 cell microsomes, with an effectiveness similar to that in microsomes prepared from identically treated 10T1/2 cells. Anti-P4501A1 had no inhibitory effect on DMBA metabolism by either induced or uninduced E041 cell microsomes. Total DMBA-metabolizing activities in BA- and TCDD-induced E041 cells were consistently 2-fold lower compared to those in similarly treated 10T1/2 cells. In addition, both induced and uninduced E041 cell microsomes formed a lower proportion of DMBA dihydrodiols relative to phenols in comparison to identically treated 10T1/2 cell microsomes (0.5 versus 1). Addition of exogenous epoxide hydrolase to E041 cell microsomes resulted in a product distribution indistinguishable from that in 10T1/2 cells. Immunoblots demonstrated 5-fold lower levels of epoxide hydrolase in E041 cell microsomes compared to 10T1/2 cell microsomes. Anti-P450EF immunoblotted a 55 kd protein in E041 cell microsomes that was induced 14-fold by BA and 6-fold by TCDD, thus paralleling the increases in the respective DMBA metabolism. It is therefore concluded that following PAH exposure endometrial stromal cells express the novel PAH-inducible mouse embryo fibroblast P450 and fail to express P4501A1. PMID- 8222048 TI - Kinetics of mammary epithelial cell proliferation in pituitary isografted BALB/c mice. AB - Recently, we have published that treatment of pituitary isografted BALB/c mice with a single injection of N-methyl-N-nitrosourea (MNU) leads to the rapid development of mammary tumors in over 90% of the animals (Guzman et al., Cancer Res., 52, 5732-5737). In the present study, we characterized the changes in proliferative activity and lobulo-alveolar differentiation of MECs at different time intervals after isografting animals with pituitary glands. Virgin BALB/c mice 1, 3, 5 or 8 weeks after pituitary isografting were either pulse-labeled for 2 h or continuously infused with bromodeoxyuridine (BrdU) and the percentage of BrdU-labeled MECs was assessed. The S-phase duration (TS) of MECs was evaluated by double labeling with [3H]thymidine and BrdU. The population potential doubling time (TP) was calculated from the values of BrdU-LI and TS. Three stages of proliferation and differentiation of MECs in pituitary isografted virgin BALB/c mice were observed: (i) A sharp increase in the percentage of proliferating MECs of the terminal ducts and ductal branchings in the first 1-2 weeks, (ii) Development of lobulo-alveolar structures from the terminal ductal and alveolar buds, between weeks 3 and 5 with the highest BrdU-LI in week 3 and (iii) Multiplication of the alveolar structures and decrease in the BrdU-LI between weeks 5 and 8. The BrdU-LIs of alveolar cells 5 weeks after isografting the animals were significantly higher than those of the ductal cells. The continuous administration of BrdU for 3, 5 or 7 days by using osmotic pumps revealed zones in the ducts where almost all MECs were labeled as well as zones lacking proliferate activity. When the BrdU administration was extended for 10-14 days, almost all (> 95%) ductal and lobular epithelial cells were labeled. A small percentage (< 5%), of ductal and lobulo-alveolar MECs cells, remained unlabeled even after 14 days infusion of BrdU. The TS and TP values were shorter in pituitary isografted animals than in controls, but no significant difference was found for either values between the ductal and alveolar cells in either isografted or control mice. Changes in proliferation kinetics of mouse MECs in pituitary isografted animals correlated with the circulating concentrations of prolactin, progesterone and 17 beta-estradiol, but not with corticosterone, growth hormone or thyroxin.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8222049 TI - Incorporation of carbon atoms from glucose into the food mutagens MeIQx and 4,8 DiMeIQx using 14C-labelled glucose in a model system. AB - Mixtures of creatinine, glucose and threonine with the addition of a small amount, 250 microCi, of [U-14C]glucose, [1-14C]glucose or [6-14C]glucose were heated at 180 degrees C for 30 min in an aqueous model system. The mixtures were purified and analysed using HPLC, scintillation and Ames tests. 2-Amino-3,8 dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-3,4,8-trimethylimidazo[4,5 f]quinoxaline (4,8-DiMeIQx) were detected as the main radioactive mutagens. The amount of MeIQx and 4,8-DiMeIQx produced from threonine was estimated at 18 and 60 nmol/mmol glucose respectively. Radioactive carbon atoms originating from glucose were also shown to be incorporated into 2-amino-3-methylimidazo[4,5 f]quinoxaline (IQx). The specific activity was calculated to be 0.6, 0.3 and 0.1 0.3 mCi/mmol for MeIQx, 4,8-DiMeIQx and IQx respectively for all three labelled forms of glucose. By the incorporation of carbon atoms originating from glucose into the imidazoquinoxaline mutagens it was clearly demonstrated that glucose is a precursor in the formation of these food mutagens. PMID- 8222050 TI - Tumor-initiating activity on mouse skin of bay region diol-epoxides of 5,6 dimethylchrysene and benzo[c]phenanthrene. AB - Previous metabolism and DNA-binding studies indicated that 5,6-dimethylchrysene (5,6-diMeC) is metabolically activated in mouse skin through formation of its 1,2 dihydrodiol (5,6-diMeC-1,2-diol) and bay region diol-epoxide (anti-5,6-diMeC-1,2 diol-3,4-epoxide). These metabolites were tested as tumor initiators on mouse skin. Included for comparison were syn-5,6-diMeC-1,2-diol-3,4-epoxide and anti 4,3-dihydroxy-2,1-epoxy- 4,3,2,1-tetrahydrobenzo[c]phenanthrene (anti-B[c]Ph-4,3 diol-2,1-epoxide). At an initiating dose of 100 nmol/mouse, 5,6-diMeC-1,2-diol and anti-5,6-diMeC-1,2-diol-3,4-epoxide were significantly more tumorigenic than 5,6-diMeC, inducing 7.1 and 3.9 skin tumors per mouse respectively compared to 1.1 induced by 5,6-diMeC. Similar results were obtained at an initiating dose of 33 nmol/mouse. This is the first example of a methylated polynuclear aromatic hydrocarbon bay region diol-epoxide which is more tumorigenic than its parent hydrocarbon on mouse skin. syn-5,6-diMeC-1,2-diol-3,4-epoxide was only weakly tumorigenic. Comparisons of anti-5,6-diMeC-1,2-diol-3,4-epoxide and anti-B[c]Ph 4,3-diol-2,1-epoxide demonstrated that the latter was a stronger tumor initiator. The results of this study confirm the bay region diol-epoxide metabolic activation pathway of 5,6-diMeC but do not provide an explanation for the relatively weak tumorigenicity of this hydrocarbon on mouse skin. PMID- 8222051 TI - DNA adduct formation in Salmonella typhimurium, cultured liver cells and in Fischer 344 rats treated with o-tolyl phosphates and their metabolites. AB - 2-Phenoxy-4H-1,3,2-benzodioxaphosphorin 2-oxide is an electrophilic and a neurotoxic metabolite of o-tolyl phosphates. In a previous paper we reported that 2-phenoxy-4H-1,3,2-benzodioxaphosphorin 2-oxide is mutagenic in Salmonella typhimurium TA100 and forms DNA adducts in incubations with nucleotides, nucleosides and isolated DNA. In the present study we compare DNA adduct formation using 32P-post-labelling assays in 2-phenoxy-4H-1,3,2 benzodioxaphosphorin 2-oxide-treated bacteria (S.typhimurium TA100) and hepatoma cells with DNA adducts formed in liver, kidney, lung and heart of tri-o-tolyl phosphate-exposed Fischer 344 male rats. In both bacteria and hepatoma cells two DNA adducts could be detected after treatment with 2-phenoxy-4H-1,3,2 benzodioxaphosphorin 2-oxide. The minor adduct co-chromatographed with synthetic N3-(o-hydroxy-benzyl)deoxyuridine 3' monophosphate after postlabelling. The major DNA adduct was a cytidine adduct, most likely N3-(o-hydroxybenzyl)deoxycytidine 3' monophosphate. Male Fischer 344 rats were treated orally for 10 days with tri o-tolyl phosphate (50 mg/kg/day) and DNA was isolated from liver, kidney, lung, heart, brain and testes 1, 4, 7 and 28 days after giving the last dose. Analysis by 32P-postlabelling revealed that two adducts were present in the DNA isolated from liver, kidney, lung and heart on the first day after giving the last dose; DNA adducts were not detected in the brain and testes. The adduct pattern after in vivo treatment with tri-o-tolyl phosphate was identical with that found in bacteria and hepatoma cells treated with 2-phenoxy-4H-1,3,2-benzo-dioxaphosphorin 2-oxide, the major adduct being N3-(o-hydroxybenzyl)deoxycytidine 3' monophosphate and the minor N3-(o-hydroxybenzyl)deoxyuridine 3' monophosphate. Both DNA adducts persisted in the lungs for the entire observation period, whereas in the kidney only the cytidine adduct could be detected 28 days after the last dose of tri-o-tolyl phosphate. In liver and heart the adducts were detectable only on the first day after completion of the treatment. The results indicate that in addition to the well established neurotoxicity, some o-tolyl phosphates may have a carcinogenic potential. PMID- 8222052 TI - Sex differences at the initiation stage of rat liver carcinogenesis--influence of growth hormone. AB - Male and female rats were initiated with a single dose of 3'-methyl-4 dimethylaminoazobenzene (3'MeDAB), N-hydroxy-2-acetylaminofluorene (N-OH-AAF) or aflatoxin B1 (AFB1) and promoted with 2-acetylaminofluorene plus partial hepatectomy (2-AAF/PH). One group of male rats in each experiment received continuous infusion of pituitary growth hormone (GH) for one week prior to initiation to feminize the secretory pattern of GH and hepatic functions. A significantly larger number of foci in male than in female rats was observed after N-OH-AAF and AFB1 initiation. Continuous GH treatment of male rats decreased the number of N-OH-AAF-initiated foci to the level in females, while no effect of GH was observed on AFB1 initiation. Initiation with 3'MeDAB showed no sex differences in number of foci. In conclusion, the present study demonstrates that sex differences occur at the initiation stage and that the secretory pattern of GH is responsible for the dimorphism in initiation with N-OH-AAF, but not with AFB1. PMID- 8222053 TI - Enzymatic reduction of chromium(VI) by human hepatic microsomes. AB - The reduction of chromium(VI) by human hepatic microsomes was investigated. The reduction rates were proportional to the amount of microsomes added and reduction was mediated by an NADPH-dependent enzymatic system which exhibited a Km for chromate of 1.04 +/- 0.18 microM and a Vmax of 5.03 +/- 0.49 nmol/min/mg protein. Relative to incubation under 0% O2, 21% O2 inhibited microsomal Cr(VI) reduction in three individuals by 53, 36 and 37%. Cr(VI) reduction was not inhibited by metyrapone, carbon monoxide, aminopyrine, piperonyl butoxide or chloroform, suggesting that cytochrome P450s did not play a major role. Thallium trichloride (0.13 and 0.26 mM), a known flavoprotein inhibitor, caused a complete inhibition of both Cr(VI) reduction and NADPH:cytochrome P450 (c) reductase activity. A partial inhibition of Cr(VI) reduction was seen in the presence of n-octylamine, which may suggest a possible role for flavin-containing monooxygenase (FMO). Overall, human microsomal Cr(VI) reduction is very different from the P450 mediated microsomal reduction observed in rodents. Specifically, the human system is much less oxygen-sensitive, has a much greater affinity for chromate and is apparently mediated by flavoproteins. PMID- 8222054 TI - 32P-postlabeling of DNA adducts of styrene-exposed lamination workers. AB - Lamination workers are exposed to high concentrations of styrene. A postlabeling method was developed for the detection of styrene-specific DNA adducts of the workers. To synthesize a standard, styrene oxide was reacted with 2' deoxyguanosine 3'-monophosphate (dGMP) and the O6-dGMP-adduct was isolated and characterized. The human samples were assayed by the nuclease P1 version of the 32P-postlabeling technique, using magnet transfer of the adducts in chromatography. The human samples were spiked with the standards to ensure identification and quantitation. In lamination workers the O6-adducts, adjusted for adduct recovery, were detected at a level of 5 adducts/10(8) nucleotides, over five times the level in the controls. PMID- 8222055 TI - Affinities for the aryl hydrocarbon receptor, potencies as aryl hydrocarbon hydroxylase inducers and relative toxicities of polychlorinated biphenyls. A congener specific approach. AB - Polychlorinated biphenyls (PCBs) are nonplanar aromatic xenobiotics that are not structurally related to polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs), yet, some PCBs are potent ligands for the aryl hydrocarbon receptor (AhR), active inducers of aryl hydrocarbon hydroxylase (AHH) and 7 ethoxyresorufin O-deethylase (EROD), and elicit toxicological responses in animals similar to PCDDs and PCDFs. We report new methodologies for quantifying the affinities of PCBs for AhR and corresponding potencies as AHH and EROD inducers. The models show that lipophilicities, electron affinities, entropies and electronic energy gaps of PCBs are key physicochemical properties controlling their AhR, AHH and EROD activities. Using 3,3',4,4'-tetrachlorobiphenyl (TCB) as the reference compound, it is shown that PCBs having higher electron affinities, lower lipophilicities and entropies than TCB are potent ligands for rat hepatic AhR. In addition, the congeners having higher binding affinities to AhR and smaller energy gaps than TCB are potent AHH and EROD inducers in rat hepatoma cells in culture. The reported models qualitatively explain and quantify AhR, AHH and EROD activities of all 209-PCBs and related xenobiotics, e.g. PCDDs and PCDFs. Furthermore, we demonstrated that AhR and AHH activities of PCBs relative to 2,3,7,8-tetrachlorodibenzo-p-dioxin correlate with corresponding in vivo relative toxicities in animals as well as assigned toxic equivalency factors. The reported methodologies are likely to be useful for identifying potentially toxic aromatic xenobiotics in mammals, and minimizing the need for animal testing. PMID- 8222056 TI - 2,3-epoxy-4-hydroxynonanal as a potential tumor-initiating agent of lipid peroxidation. AB - Trans-4-hydroxy-2-nonenal (HNE) is a product of lipid peroxidation. In the presence of t-butyl hydroperoxide the racemic HNE readily converts to its epoxide, 2,3-epoxy-4-hydroxynonanal (EH), as a pair of diastereomers. In this study, the potential roles of HNE and EH as tumor initiating agents were assessed. The mutagenicities of HNE and EH isomers in Salmonella strains TA100 and 104 were examined. In addition, the tumor initiating activities of HNE and EH were evaluated in bioassays involving either topical application in CD-1 mice or i.p. administration in newborn CD-1 mice. In the mutagenicity assays, EH isomers induced similar levels of revertants in both tester strains, although EG isomers were previously shown to react with bases in DNA with different specificity (Sodum, R.S. and Chung, F.-L., Cancer Res., 51, 137-143, 1991). The major isomer induced approximately 20,000 revertants/mumol in TA100 and 15,000 revertants/mumol in TA104, whereas, the minor isomer induced approximately 40,000 revertants/mumol in TA100 and 20,000 revertants/mumol in TA104. HNE was, however, not mutagenic under the assay conditions. In the tumor bioassays, EH was a weak tumorigen in CD-1 mice upon topical application followed by TPA promotion, yielding 0.55 tumors/mouse and 40% tumor incidence at a total dose of 128 mumol/mouse versus 0.02 tumors/mouse and 5% tumor incidence in the control group. Both HNE and EH induced liver tumors in male mice, but not in female mice. However, the incidences were not statistically significant. EH administered i.p. at a total dose of 200 nmol/mouse exacerbated the chronic spontaneous nephropathy in newborn CD-1 mice. Although the incidence of mild nephropathy was comparable in both EH-treated and control groups, the incidence of more severe lesions in mice treated with 200 nmol/mouse was 21%; while it was 0% in the control group. Furthermore, two mice at each dose level of EH showed a tubule profile with complex hyperplastic lining, suggestive of atypical hyperplasia. Again, HNE was not as active as EH in these bioassays. These results suggest a possible role of EH in tumorigenesis associated with lipid peroxidation. PMID- 8222057 TI - Allyl isothiocyanate is selectively toxic to transformed cells of the human colorectal tumour line HT29. AB - Allyl isothiocyanate, a constituent of mustard and certain vegetables found in the human diet, was tested for cytotoxic and cytostatic effects in HT29 human colon carcinoma cells in vitro. For an exposure time of 24 h, allyl isothiocyanate exhibited a Dq of 0.32 microgram/ml and a D0 of 0.74 micrograms/ml. Following detransformation of the cells by treatment with sodium butyrate or dimethylformamide the cells became more resistant to the cytotoxic effects of allyl isothiocyanate, the Dq increasing to 0.74 microgram/ml and the D0 to 0.96 microgram/ml (with butyrate) or 0.84 microgram/ml (with dimethylformamide). At the Dq value for detransformed cells the survival of the control cells was reduced to 56%. Allyl isothiocyanate was also found to be less cytostatic to the mass growth of detransformed populations in that daily doses of 1.6 micrograms/ml over a week reduced the final number of detransformed cells relative to untreated cultures by < 25% whilst growth of the transformed cultures was reduced by > 60%. Given this increased sensitivity of the cells to allyl isothiocyanate when in the transformed state, it is hypothesized that, when consumed in the human diet, this compound may protect against the development of colorectal cancer by selectively inhibiting the growth of transformed cell clones within the gastrointestinal mucosa. PMID- 8222058 TI - Heterologous gap junctional intercellular communication in normal and morphologically transformed colonies of Syrian hamster embryo cells. AB - A study was made of whether normal and morphologically transformed colonies in the Syrian hamster embryo (SHE) cell transformation assay performed heterologous gap junctional intercellular communication (GJIC). Two compounds, 12-O tetradecanoylphorbol-13-acetate (TPA) and Na-orthovanadate (vanadate), which induce high frequencies of morphological transformation in SHE cells, have been employed. Three approaches were used to study the possibility of heterologous GJIC. (i) Morphologically transformed colonies (induced by TPA) partially overlapping with normal colonies were selected. Cells in the border area were micro-injected with the fluorescent dye Lucifer yellow to determine whether the dye spread to cells belonging to the other colony. This approach proved to be unsuccessful due to an inability to pinpoint which cells belonged to which colony. (ii) X-irradiated, non-dividing feeder cells are easily recognized by their large size. Feeder cells in contact with normal or TPA-transformed colonies were injected with Lucifer yellow. The dye was found to spread to most of the contacting cells, irrespective of whether they belonged to a normal or morphologically transformed colony. (iii) TPA- and vanadate-exposed colonies were labelled by endocytosis of Lucifer yellow overnight. This resulted in a punctate fluorescent pattern. Unlabelled, previously unexposed cells were seeded onto the dishes and incubated for 3.5-7 h. The ability to perform heterologous GJIC between the newly seeded cells and labelled colony cells was investigated. Both normal and transformed colonies were found to be able to communicate with the newly seeded cells. Thus, the present results indicate that selective communication is not a general property of morphologically transformed SHE cell colonies. PMID- 8222059 TI - Enzymatic phase II activation of the N-hydroxylamines of IQ, MeIQx and PhIP by various organs of monkeys and rats. AB - 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) are mutagenic and carcinogenic heterocyclic amines produced during the ordinary cooking of meat. These compounds undergo metabolic activation via both cytochrome P450-mediated N-oxidation and phase II esterification in order to exert their genotoxicity. In the current study, we examined the in vitro phase II activation of N-hydroxy-IQ, N-hydroxy-PhIP and N-hydroxy-MeIQx by cytosolic acetyltransferase, sulfotransferase, aminoacyl-tRNA synthetase and phosphatase from a number of tissues including liver, kidney, colon and heart. These tissues were chosen for study because each is either a target organ for carcinogenicity or has displayed high levels of DNA adducts in in vivo studies with the heterocyclic amines. Cytosol from various tissues of both monkeys and rats was incubated with and without the respective cofactors, and carcinogen binding to calf thymus DNA was measured by 32P-postlabeling analysis. Our results show that all four phase II enzymes may participate in the activation of the N hydroxylamines. However, the degree of activation depends on the substrate, tissue and animal species. For example, in both monkeys and rats, the highest acetyl CoA-enhanced binding was observed with N-hydroxy-IQ and the lowest acetyl CoA-enhanced binding was observed with N-hydroxy-MeIQx. In contrast, no significant adenosine 3'-phosphate 5'-phosphosulfate-dependent activation of N hydroxy-IQ was observed with monkey cytosol from liver, kidney, heart or colon but the sulfotransferase-mediated activation of N-hydroxy-PhIP was at least 10 times higher in all four tissues of monkeys than in rats. Prolylation appears important in the activation of all three N-hydroxylamines by rat liver and heart cytosol, whereas in monkeys, prolylation appears important in kidney cytosol. The differences observed in the phase II activation of heterocyclic amines may have implications for DNA adduct formation, toxicity and carcinogenicity. PMID- 8222060 TI - Genetic consequences of tolerance to methylation DNA damage in mammalian cells. AB - We previously characterized a clone of CHO cells, clone B, that displayed tolerance to the cytotoxic effects of N-methylnitrosourea (MNU) and 6-thioguanine (6-TG). To determine whether this phenotype affected the mutagenic response of the cells, MNU-induced mutation to 8-azaadenine resistance (8-AAr) was measured in the parental and clone B cells. Comparable mutation frequencies were found in the two cell lines up to 0.5 mM MNU, while at higher MNU concentrations mutations could be reproducibly measured only in clone B cells. Similar amounts of DNA methylated bases were found in the two cell lines after a 30 min treatment with different concentrations of [3H]MNU and the same linear relationship was observed when mutation induction by MNU was plotted as a function of the amount of O6 methylguanine (O6-MeGua) in DNA, indicating that mutation induction in both cell lines was related to the presence of this methylated base. Fifteen MNU-induced 8 AAr mutants were isolated from each cell line and the sequences of the adenine phosphoribosyltransferase (aprt) mutations determined. The type (in 90% of the cases, GC to AT transitions), the sequence context and the strand localization of the mutations indicated that all mutations were targeted at O6-MeGua in DNA and no difference was found between the two lines. These results are consistent with a mechanism of tolerance of O6-MeGua that does not alter the processing of this methylated base into a mutation. Growth in 6-TG induced point mutations in clone B but not in the parental cells. A model is proposed in which the alkylation tolerant variant is altered in a mismatch correction pathway responsible for the cytotoxicity of the methylated base. PMID- 8222061 TI - Methyl methanesulfonate adduct formation and repair in the DHFR gene and in mitochondrial DNA in hamster cells. AB - Chinese hamster ovary CHO-B11 cells were exposed to methyl methanesulfonate (MMS) and the formation and repair of N-methylpurines were measured in the endogenous dihydrofolate reductase (DHFR) gene domain and in mitochondrial DNA by alkaline hydrolysis which generates strand breaks at apurinic (AP) sites formed after neutral depurination. The initial levels of damage in the transcriptionally active DHFR gene, 3'-flanking non-transcribed region and in mitochondrial DNA were slightly different; the highest level of damage was in mitochondrial DNA, the lowest in the DHFR gene. The rate of adduct removal was similar in all three DNA regions examined. We conclude that there is no preferential repair of MMS induced N-methylpurines under these conditions, and that these lesions are efficiently removed from mitochondrial DNA. PMID- 8222062 TI - Trans-1,2-dihydro-1,2-dihydroxy-6-aminochrysene is metabolized to form a major adduct with deoxyguanosine and produces mutations in the hprt gene of Chinese hamster ovary cells at G:C basepairs. AB - 6-Nitrochrysene can be activated to genotoxic derivatives by two major metabolic pathways: nitroreduction to N-hydroxy-6-aminochrysene, and a combination of ring oxidation and nitroreduction that involves the intermediate formation of trans 1,2-dihydro-1,2-dihydroxy-6-aminochrysene (6-AC-1,2-dihydrodiol). The DNA adduct formed from this latter pathway was evaluated by reacting individual deoxynucleoside 5'-monophosphates with 6-AC-1,2-dihydrodiol in the presence of liver microsomal enzymes from 3-methylcholanthrene-pretreated rats. Binding was greatest to deoxyguanosine monophosphate and the major deoxyguanosine (dG) adduct co-chromatographed with the single major adduct formed from the microsome catalyzed reaction of 6-AC-1,2-dihydrodiol with DNA. In order to characterize the mutational changes associated with the 6-AC-1,2-dihydrodiol pathway, we analyzed the mutational spectrum produced by 6-AC-1,2-dihydrodiol in the hypoxanthine guanine phosphoribosyltransferase (hprt) gene of CHO-K1 cells. cDNA was synthesized from the RNA of 28 6-thioguanine-resistant mutants, the hprt coding region amplified by the polymerase chain reaction, and the DNA products directly sequenced. Twenty independent primary mutations were found: 12 G:C-->T:A transversions, three G:C-->C:G transversions, one G:C-->A:T transition, one A:T- >T:A transversion, two -1 frameshift mutations in sequences containing consecutive guanines, and one 11 bp deletion. All G:C basepair substitutions had the mutated dG on the non-transcribed strand and 86% of the G:C basepair substitutions had one purine 3' to the mutated dG. The pattern of 6-AC-1,2 dihydrodiol-induced basepair substitutions was distinct from the pattern observed in solvent control mutants. These results are consistent with the formation of a promutagenic dG adduct from a metabolite of 6-AC-1,2-dihydrodiol. PMID- 8222063 TI - Effects of benzo[a]pyrene and (+-)-trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene on mitosis in Chinese hamster V79 cells with stable expression of rat cytochrome P4501A1 or 1A2. AB - The effect of bioactivation of benzo[a]pyrene (B[a]P) and (+-)-trans-7,8 dihydroxy-7,8-dihydrobenzo[a]pyrene (B[a]P-7,8-diol) on spindle disturbances and toxicity has been investigated in V79 Chinese hamster cells genetically engineered to express cytochrome P4501A1 (CYP1A1) and cytochrome P4501A2 (CYP1A2). B[a]P induces spindle disturbances in native V79 Chinese hamster cells. This effect was enhanced by the expression of CYP1A1 but not CYP1A2. The increased effect seen in the CYP1A1-expressing cell line could be brought back to the level seen in the native cell line by alpha-naphthoflavone in a dose dependent manner. This strongly suggests that a CYP1A1-dependent metabolite, conceivably (+-)-trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene (BPDE) accounts for the increased spindle disturbing effect. B[a]P-7,8-diol induced spindle disturbances at remarkably low concentrations, 10(-8) M, irrespective of expression of the two CYPs. Our data suggest that B[a]P-7,8-diol is the most potent spindle-disturbing metabolite, whereas BPDE is the most important metabolite concerning mutagenesis. The concentrations inducing spindle disturbances correspond to those that are positive in mutation assays. We hypothesize that B[a]P is a complete carcinogen because of its ability to induce both aneuploidy and mutations after metabolic conversion of low non-cytotoxic concentrations. PMID- 8222064 TI - p53 mutations in phenacetin-associated human urothelial carcinomas. AB - Chronic abuse of the analgesic drug phenacetin is associated with an increased risk of development of transitional cell carcinomas of the urinary tract. It is unclear whether phenacetin acts through chronic tissue damage (phenacetin nephropathy) or via a genotoxic metabolite causing promutagenic DNA lesions. In the present study, we investigated 15 urothelial carcinomas from 13 patients with evidence of phenacetin abuse. Tumors were screened for p53 mutations in exons 5-8 by single-strand conformation polymorphism (SSCP) analysis, followed by direct sequencing of PCR-amplified DNA. p53 Mutations were detected in 8/14 primary tumors (57%). All except one were missense mutations located in exon 5 (three mutations), exon 6 (one), exon 7 (two) and exon 8 (one). The type of mutation varied, with a preference for CpG sites. A frameshift mutation resulting from the insertion of a single cytosine at codons 151/152 was detected in a bladder tumor and its lung metastasis. Urothelial carcinomas located in the renal pelvis and in the ureter of the same patient exhibited two different mutations, strongly suggesting that they developed independently. Another patient had tumors in the renal pelvis and bladder, both of which contained the same p53 mutation, indicating intracavitary metastatic spread. This demonstrates that screening of p53 mutations allows the clonal origin of tumors in patients with multiple primary and metastatic lesions to be determined. None of the tumors investigated contained mutations in codons 12, 13 or 61 of H-ras or K-ras protooncogenes. PMID- 8222065 TI - The heterocyclic amines IQ and MeIQx show no promotive effect in a short-term in vivo liver carcinogenesis assay. AB - The food-derived heterocyclic amines 2-amino-3-methyl-imidazo[4,5-f]quinoline (IQ) and 2-amino-3,8-dimethyl-imidazo[4,5-f]quinoxaline (MeIQx) are carcinogenic in rodents as well as in non-human primates (IQ). Using a short-term liver carcinogenesis model the compounds were found to be only weak initiators. It is also possible that these amines are involved in other phases of the carcinogenesis process. In an attempt to study the promotive effects of IQ and MeIQx, these heterocyclic amines were repeatedly given by gavage, over a period of 10 days, to Wistar rats previously initiated with diethylnitrosamine (DEN) and subjected to a partial hepatectomy. The rats were killed 1 month after DEN administration and their livers examined for glutathione-S-transferase positive (GST-P) foci. The positive control, 2-acetylaminofluorene, significantly increased the development of GST-P foci in the DEN-treated rats compared to the negative control, saline. However, neither the IQ- nor the MeIQx-treated groups differed from the controls and thus these amines do not promote the growth of DEN initiated GST-P liver foci in this model. PMID- 8222066 TI - The effect of L-carnitine and acetyl-L-carnitine on the disappearance of DNA single-strand breaks in human peripheral blood lymphocytes. AB - DNA single-strand breaks (SSBs) and their disappearance during repair incubation were determined by alkaline filter elution in freshly isolated human peripheral blood lymphocytes (PBLs) after in vitro treatment with either the oxygen radical generating system of xanthine oxidase (XOD) plus hypoxanthine (HYP) or the alkylating agent N-ethyl-N'-nitrosourea (ENU). The elution curves obtained with DNA from PBLs treated with XOD/HYP were markedly nonlinear, possibly as a result of a nonrandom induction of SSBs along the DNA strands. The disappearance of XOD/HYP-induced SSBs during the initial repair period was quite slow; only 20 +/- 7% (n = 6) of the induced SSBs had disappeared after a 2 1/2 h repair incubation. However, by 24 h the elution curves obtained with DNA from treated PBLs were indistinguishable from those obtained with DNA from nontreated control cells, indicating complete repair. Treatment of PBLs with ENU resulted in linear elution curves. Approximately 50% of the total amount of ENU-induced SSBs had disappeared within 1 h in PBLs from most donors; the additional SSBs were found to be persistent (Beorrigter, M.E.T.I., Mullaart, E., Berends, F., and Vijg, J. (1991) Induction and disappearance of DNA strand breaks and/or alkali-labile sites in human lymphocytes exposed to N-ethyl-N'-nitrosoureas. Carcinogenesis, 12, 77-82). Preincubation of PBLs with 5 mM L-carnitine, a trans-mitochondrial carrier of acetyl and long-chain acyl groups, or 5 mM acetyl-L-Carnitine, resulted in a more rapid disappearance of XOD/HYP-induced SSBs (48 +/- 23% and 48 +/- 30% respectively). Preincubation of PBLs with different doses of L-carnitine, before exposure to 0.5 mM ENU, increased SSB disappearance dependent on the dose and donor PBLs. In conclusion, these studies suggest that treatment with L-carnitine accelerates the disappearance of SSBs induced by oxygen radicals and alkylating agents. PMID- 8222067 TI - Promotion of preneoplastic lesions and induction of CYP2B by unleaded gasoline vapor in female B6C3F1 mouse liver. AB - An initiation-promotion protocol was used to test the hypothesis that unleaded gasoline (UG) vapor acts as a liver tumor promoter in female mice under exposure conditions in which UG was hepatocarcinogenic in a cancer bioassay. Twelve day old female B6C3F1 mice were injected with N-nitrosodiethylamine (DEN, 5 mg/kg, i.p.) or vehicle. Starting at 5-7 weeks of age, mice were exposed by inhalation 6 h/day, 5 days/week for 13 weeks to 0 or 2039 p.p.m. of PS-6 blend UG, the same gasoline blend used in the cancer bioassay. Putative preneoplastic lesions in liver, characterized mainly as basophilic foci in H&E-stained liver sections, were found exclusively in mice treated with DEN. While similar numbers of altered hepatic foci were found in DEN-initiated mice treated with 0 or 2039 p.p.m. UG, UG treatment significantly increased both the mean volume (3.2-fold) and the volume fraction (3.6-fold) of the foci. To determine if UG induced CYP2B, a subfamily of cytochrome P450 commonly induced by liver tumor promoters in rodents, pentoxyresorufin-O-dealkylase (PROD) activity was assayed in hepatic microsomes derived from the above livers. UG vapor increased hepatic PROD activity approximately 8-fold, while increasing cytochrome P450 content only approximately 30%. To ascertain if a more recent blend of UG, API 91-1, would have similar biological effects as PS-6, female B6C3F1 mice were gavaged for 3 days with corn oil or 1800 mg/kg/day PS-6 or API 91-1 blend UG. PS-6 and API 91-1 blend UG induced similar increases in relative liver weight (approximately 25%), PROD activity (approximately 9-fold) and hepatocyte labeling index (approximately 8-fold) relative to controls. These data demonstrate that PS-6 blend UG vapor promotes preneoplastic lesions and induces CYP2B in female mouse liver under exposure conditions in which it causes liver tumors, and suggest that a more recent blend of UG may have similar effects. PMID- 8222068 TI - 32P-postlabelling analysis of DNA adducts of 4,4'-methylenebis(2-chloroaniline) in target and nontarget tissues in the dog and their implications for human risk assessment. AB - 4,4'-Methylenebis(2-chloroaniline) (MOCA) has considerable human occupational exposure and it induces urinary bladder tumors in the dog, a species that has been often used as a model for aromatic amine-induced urinary bladder carcinogenesis in humans. Metabolic activation and formation of DNA adducts are considered to be critical steps in this process; and two major C8-adenine adducts have been shown to be formed in vitro by reaction with the proximate carcinogenic metabolite N-hydroxy-MOCA. MOCA-DNA adducts have also been detected in vivo in treated rats and in exfoliated urothelium of a worker accidentally exposed to MOCA. Thus, the aim of this study was to detect and quantify DNA adducts in the urinary bladder of dogs exposed to MOCA and thereby provide data that could be useful for risk assessment after human exposure to MOCA. Beagle dogs were treated with single and multiple doses of MOCA and DNA adduct levels were determined in liver and bladder epithelium. After a single dose, adduct levels in the liver were 1.5-fold higher than that in the bladder epithelium. Adduct levels in these two organs increased 3- to 5-fold after 10 doses and adducts in the liver were then 2.8-fold higher than that in the bladder epithelium. The levels found in these two organs after single exposures were compared, per unit exposure dose, with that reported for other carcinogenic aromatic amines. The comparison showed that MOCA was as effective in DNA adduct formation as most other potent urinary bladder carcinogens. These results suggest that MOCA may have high carcinogenic potential in humans and are consistent with the recent classification of MOCA as a probable human carcinogen. PMID- 8222069 TI - Promotive effects of diethylstilbestrol, its metabolite (Z,Z-dienestrol) and a stereoisomer of the metabolite (E,E-dienestrol) in tumorigenesis of rat mammary glands pregnancy-dependently initiated with radiation. AB - Wistar-MS rats received whole body irradiation with 260 cGy gamma-rays at day 20 of pregnancy and then were treated with diethylstilbestrol (DES), E,E-dienestrol (E,E-DIES) or Z,Z-dienestrol (Z,Z-DIES) for 1 year. DES administration caused the highest incidence of mammary tumors with a concomitant reduction of gain in body weight. When E,E-DIES or Z,Z-DIES in pellet form was implanted, the incidence of tumors was significantly lower than that observed in rats treated with DES. To clarify the increased susceptibility to mammary tumorigenesis after DES administration we measured hormone levels in the serum of rats implanted with pellets containing derivatives of the synthetic estrogens. The serum prolactin concentration was significantly increased by DES administration. When E,E-DIES or Z,Z-DIES pellets were implanted the prolactin level was markedly reduced to 4.5% and 0.7% of that observed in DES-treated rats, respectively. In addition, the serum concentrations of estradiol-17 beta and progesterone in rats with Z,Z-DIES pellets were higher than those of rats with DES or E,E-DIES pellets. A large number of DES-induced mammary tumors were positive for both estrogen and progesterone receptors, but no tumors negative for both receptors were obtained. The findings suggest that DES acts directly on radiation-initiated mammary cells via binding with estrogen receptors and/or stimulates the secretion of prolactin from the pituitary glands. PMID- 8222070 TI - Identification of N2-(deoxyguanosin-8-yl)-2-amino-3,8-dimethyl-imidazo[4,5- f]quinoxaline 3',5'-diphosphate, a major DNA adduct, detected by nuclease P1 modification of the 32P-postlabeling method, in the liver of rats fed MeIQx. AB - The carcinogenic heterocyclic amine 2-amino-3,8-dimethyl-imidazo[4,5 f]quinoxaline (MeIQx) is widely distributed in cooked foods. The nuclease P1 method increased the sensitivity of the standard 32P-postlabeling analysis about 1000-fold for detection of MeIQx-DNA adducts. The recovery of MeIQx-DNA adducts by the nuclease P1 method was determined to be about 50% using liver DNA of a rat treated with [14C]MeIQx intragastrically. By the nuclease P1 method five adducts were detected in the liver DNA of rats fed MeIQx and two of them, including the most abundant one, were identified as MeIQx-deoxyguanosine adducts by comparison with the adducts formed in in vitro reactions of N-acetoxy-2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline with the four 2'-deoxyribonucleotides. The most abundant adduct in vivo was identified as N2-(deoxyguanosin-8-yl)-MeIQx 3',5' diphosphate (3',5'-pdGp-C8-MeIQx). MeIQx-DNA adduct levels in human tissues could be determined by the nuclease P1 modification of the 32P-postlabeling method in combination with HPLC, and thus provide information on the roles of MeIQx in human carcinogenesis. PMID- 8222071 TI - Cisplatin sensitivity/resistance in UV repair-deficient Chinese hamster ovary cells of complementation groups 1 and 3. AB - We assessed the possible role of the human repair genes, ERCC1 and ERCC3, in resistance to cisplatin-induced cytotoxicity. The UV repair-deficient Chinese hamster ovary (CHO) 43:3B [designated ERCC1(-)] cell line and its paired subline 83-J5, which is stably transfected with the human DNA excision repair gene ERCC1 [designated ERCC1(+)], were used in this study. UV repair-deficient CHO 27-1 cells [designated ERCC3(-)] and its paired subline designated 'ERCC3(+)', which is stably transfected with the human DNA excision repair gene ERCC3, were also used. In each pair of cell lines, we assessed cisplatin cytotoxicity, cellular drug accumulation and platinum-DNA adduct repair after 1 h drug exposures. Drug accumulation and DNA repair were assessed by atomic absorption spectrometry with Zeeman background correction. ERCC1(+) cells (IC50 = 4.0 microM) were 5-fold more resistant to cisplatin than ERCC1(-) cells (IC50 = 0.75 microM). ERCC1(+) cells repaired 25% of DNA lesions in cellular DNA within a 6 h time period following an IC50 drug exposure and repaired 48% over 24 h. No DNA repair was observed in ERCC1(-) cells during the same time periods. Both cell lines showed similar patterns of drug accumulation. For ERCC3(-) cells (IC50 = 54 microM) and ERCC3(+) cells (IC50 = 49 microM), the profiles of cisplatin sensitivity and cellular drug accumulation were similar. When treated with 50 microM cisplatin, these cells showed similar patterns of drug accumulation, and were equally efficient at forming and repairing lesions in cellular DNA. These data show that in UV repair deficient CHO cells, ERCC1 confers resistance to cisplatin and confers the ability to remove platinum from cellular DNA. In contrast, ERCC3 does not influence cisplatin drug sensitivity or adduct repair capability. This suggests that ERCC1 may be a determinant of cisplatin resistance, whereas ERCC3 is probably not. PMID- 8222072 TI - A mutational hot spot induced by N-hydroxy-aminofluorene in dihydrofolate reductase mutants of Chinese hamster ovary cells. AB - Eighteen mutants deficient in dihydrofolate reductase (DHFR) activity were induced with 0.5 microM N-hydroxy-aminofluorene in four separate experiments. This carcinogen dose killed approximately 80% of the treated cells and resulted in a mutational frequency approximately 3 x 10(-6). The nature of the induced changes in each of the mutants was determined by direct sequencing following polymerase chain reaction amplification, or in one instance, by Southern blot analysis. Nearly all (15/17) of the mutations were single base changes. Consistent with the binding specificity of this chemical, all mutations were targeted to guanine bases. The predominant change was G:C-->T:A transversion which was evident in 11/15 mutants. A single dG-AF mutational hotspot was noted at a site in the DHFR coding sequence of exon 4; one-third of the induced point mutations arose at this position. These results are compared with our previous analyses of mutants induced with the related aromatic amine, N-2-acetoxy-2-acetyl aminofluorene. PMID- 8222073 TI - Post-mortem stability of benzo[a]pyrene diolepoxide--DNA adducts in rat organs. AB - In order to evaluate the stability of benzo[a]pyrene diolepoxide--DNA adducts two separate studies were carried out in rats, either treated i.p. with benzo[a]pyrene (100 mg/kg body wt) or sham-exposed. The measurement of DNA adducts in 155 samples of liver, lung or heart, each of them tested in duplicate, was performed by synchronous fluorescence spectrophotometry. In the first study fragments of rat liver or lung were stored for varying times at varying temperatures. No decrease of adduct levels occurred at 4 degrees C for at least 72 h, whereas a significant decrease was recorded in both liver and lung after 48 h at 20 degrees C or 24 h at 37 degrees C. In the second study liver, lungs and heart were collected from rats either immediately after killing or after storage of cadavers for 16 h at 20 degrees C or 16 h at 20 degrees C plus 24 h at 4 degrees C, thereby mimicking typical storage conditions of human cadavers before autopsy. Under these conditions no significant variation of fluorescent adducts was observed in any organ. In conclusion, at least for this kind of adduct, the use of autopsy samples following proper storage of cadavers seems to be acceptable. PMID- 8222074 TI - Overexpression of messenger RNA for cholecystokinin-A receptor and novel expression of messenger RNA for gastrin (cholecystokinin-B) receptor in azaserine induced rat pancreatic carcinoma. AB - Using receptor binding assays, we have previously demonstrated the overexpression of the high-affinity cholecystokinin (CCK) receptor and the novel expression of the gastrin (CCK-B) receptor in the azaserine-induced rat pancreatic carcinoma DSL-6. Since cDNA of both the CCK-A receptor (classical pancreatic CCK receptor) coding region and the CCK-B receptor coding region have recently been cloned and sequenced, we investigated the expression of messenger RNA of these receptors in DSL-6 pancreatic carcinoma. Our results showed that the 32P-labelled cDNA probe of the CCK-A receptor coding region hybridized with an approximately 2.7 kb mRNA from both DSL-6 pancreatic carcinoma and normal rat pancreas. However, the relative expression of the CCK-A receptor mRNA in DSL-6 pancreatic carcinoma was approximately 8-fold of that in normal rat pancreas. The 32P-labelled cDNA probe of the CCK-B receptor coding region hybridized with an approximately 2.7 kb mRNA from DSL-6 pancreatic carcinoma; no hybridizing mRNA could be identified from normal rat pancreas. In summary, the CCK-A receptor mRNA is overexpressed approximately 8-fold and the gastrin (CCK-B) receptor mRNA is novelly expressed in DSL-6 pancreatic carcinoma as compared to normal rat pancreas. These results further confirm our previous findings based on receptor binding assays. The gene overexpression of the CCK-A receptor and the novel gene expression of the gastrin (CCK-B) receptor may be generated by alterations in gene regulation during carcinogenesis, and may play an important role in promoting tumor growth. PMID- 8222075 TI - Platelets from spontaneously hypertensive rats exhibit decreased expression of inhibitory guanine nucleotide regulatory protein. Relation with adenylyl cyclase activity. AB - We have recently demonstrated an enhanced expression of inhibitory guanine nucleotide regulatory protein (Gi) in the heart and aorta from spontaneously hypertensive rats (SHR) as compared with control Wistar-Kyoto (WKY) rats; this enhanced Gi expression was associated with an increased inhibition of adenylyl cyclase by inhibitory hormones and decreased stimulation of adenylyl cyclase by stimulatory hormones. In the present studies, we have determined the levels of stimulatory and inhibitory guanine nucleotide regulatory proteins (Gs and Gi, respectively) in platelets from SHR by cholera toxin- and pertussis toxin catalyzed ADP-ribosylations, respectively, as well as by immunoblotting techniques using specific antibodies for Gs and Gi. Cholera toxin catalyzed the ADP-ribosylation of a single protein of M(r) 45,000 in rat platelets from SHR and WKY rats, and the labeling of this band was not altered in SHR as compared with WKY rats. Pertussis toxin, on the other hand, catalyzed the ADP-ribosylation of a single protein band of M(r) 41,000 in platelets from SHR and WKY rats, and unlike the response in heart and aorta, the labeling of this band was significantly decreased in SHR as compared with WKY rats. Furthermore, immunoblotting experiments using AS/7 antibody, which is specific for Gi alpha-1 and Gi alpha-2, showed a decrease in Gi alpha-2 in platelets from SHR as compared with WKY rats. In addition, the inhibitory effects of angiotensin II and atrial natriuretic factor on adenylyl cyclase and cAMP levels were completely abolished in SHR platelets, whereas the stimulatory effects of GTP, N-ethylcarboxamide adenosine, prostaglandin E1, and forskolin on adenylyl cyclase and cAMP levels were enhanced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222076 TI - Assessment of smooth muscle contribution to descending thoracic aortic elastic mechanics in conscious dogs. AB - Early investigators found contradictory evidence that vascular smooth muscle activation reduces the elastic modulus of the arterial wall under isotonic conditions but increases it under isometric conditions, concomitant with increased pulse-wave velocity. We examined the individual contributions of aortic constituents to the elastic modulus of the aortic wall to determine if isobaric analysis produces an accurate assessment of vascular smooth muscle activation. We used a modified Maxwell model assuming an incremental elastic modulus (Einc) composed of the elastic modulus of elastin fibers (EE), the elastic modulus of collagen fibers (EC) affected by the fraction of collagen fibers (fC) recruited to support wall stress, and the elastic modulus of the vascular smooth muscle (ESM) according to the following formula: Einc = EE+EC x fC+ESM.Einc was assessed in eight conscious dogs using descending thoracic aortic pressure (microtransducer) and diameter (sonomicrometry) measurements. Stress-strain relations in the control state and during activation of smooth muscle by continuous administration of phenylephrine (5 micrograms.kg-1 x min-1) were obtained by transient occlusions of the descending aorta and inferior vena cava. Results were as follows: EE was 4.99 +/- 1.58 x 10(6) dynes/cm2 (mean +/- SD), and EC was 965.8 +/- 399.8 x 10(6) dynes/cm2, assessed during the control state. Phenylephrine administration increased the theoretical pulse-wave velocity (Moens Korteweg equation) from 5.25 +/- 1.03 m/s during the control state to 7.57 +/- 2.53 m/s (P < .005). Active muscle exhibited a unimodal stress-strain curve with a maximum stress of 0.949 +/- 0.57 x 10(6) dynes/cm2 at a corresponding strain value of 1.299 +/- 0.083. The maximum value observed corresponded, on the pressure-diameter curve of the active artery, to a pressure of 234.28 +/- 46.6 mm Hg and a diameter of 17.94 +/- 1.6 mm. The maximum ESM derived from the stress strain relation of the active muscle was 8.345 +/- 7.56 x 10(6) dynes/cm2 at a strain value of 1.283 +/- 0.079. This point was located at 208.01 +/- 40.8 mm Hg and 17.73 +/- 1.41 mm on the active pressure-diameter curve. During activation of vascular smooth muscle, Einc decreased (P < .05) when plotted against internal pressure but increased (P < .05) when plotted against strain, over the operative range.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8222077 TI - Vascular cell-derived heparan sulfate shows coupled inhibition of basic fibroblast growth factor binding and mitogenesis in vascular smooth muscle cells. AB - Basic fibroblast growth factor (bFGF) has been previously shown to be mitogenic for vascular smooth muscle cells (VSMCs) in vivo, but only after vascular injury. We show in the present study that the regulation of bFGF-stimulated VSMC proliferation, by vascular cell-secreted heparin-like compounds, correlates with inhibition of bFGF binding to cell-associated heparin sulfate proteoglycans. The stimulation of cultured VSMC proliferation by bFGF was markedly reduced when these cells were cocultured with confluent endothelial cells or confluent VSMCs (100.8 +/- 8.4% and 55.6 +/- 2.3% inhibition, respectively) or with conditioned media from these two cell types. Balb/c3T3 fibroblasts had no statistically significant effect on bFGF-stimulated VSMC proliferation. Vascular cell conditioned media also inhibited bFGF binding to heparan sulfate proteoglycans on VSMCs, and the inhibition of binding correlated linearly with the inhibition of proliferation after a critical amount of binding was inhibited (44%) (r = .952, P < .0001). Heparinase or heparitinase treatment of conditioned media removed the bFGF-inhibitory effects, presumably by degrading heparin-like compounds. Indeed, heparin itself mimicked the inhibitory effects of conditioned media on bFGF mediated proliferation and binding to heparin sulfate proteoglycans. These results suggest a bFGF regulatory role for vascular cell-produced heparin-like compounds, linking the mitogenic effects with binding to heparan sulfate proteoglycans for this heparin-binding growth factor. PMID- 8222078 TI - Sustained depolarization-induced outward current in human atrial myocytes. Evidence for a novel delayed rectifier K+ current similar to Kv1.5 cloned channel currents. AB - Depolarization of human atrial myocytes activates a transient outward current that rapidly inactivates, leaving a sustained outward current after continued depolarization. To evaluate the ionic mechanism underlying this sustained current (Isus), we applied whole-cell voltage-clamp techniques to single myocytes isolated from right atrial specimens obtained from patients undergoing coronary bypass surgery. The magnitude of Isus was constant for up to 10 seconds at +30 mV and was unaffected by 40 mmol/L tetraethylammonium, 100 nmol/L dendrotoxin, 1 mmol/L Ba2+, 0.1 mumol/L atropine, or removal of Cl- in the superfusate. Isus could be distinguished from the 4-aminopyridine (4AP)-sensitive transient outward current (Ito1) by differences in voltage-dependent inactivation (1000-millisecond prepulse to -20 mV reduced Ito1 by 91.7 +/- 0.1% [mean +/- SEM], P < .001, versus 9.4 +/- 0.4% reduction of Isus) and 4AP sensitivity (IC50 for block of Ito1, 1.96 mmol/L; for Isus, 49 mumol/L). Isus activation had a voltage threshold near -30 mV, a half-activation voltage of -4.3 mV, and a slope factor of 8.0 mV. Isus was not inactivated by 1000-millisecond prepulses but was reduced by 16 +/- 8% (P < .05) at a holding potential of -20 mV relative to values at a holding potential of -80 mV. Isus activated very rapidly, with time constants (tau) at 25 degrees C ranging from 18.2 +/- 1.8 to 2.1 +/- 0.2 milliseconds at -10 to +50 mV, two orders of magnitude faster than previously described kinetics of the rapid component of the delayed rectifier K+ current. At 16 degrees C, Isus activation was greatly slowed (tau at +10 mV, 46.7 +/- 4.1 milliseconds; tau at 25 degrees C, 7.1 +/- 0.8 milliseconds; P < .01), and the envelope of tails test was satisfied. The reversal potential of Isus tail currents changed linearly with log [K+]o (slope, 55.3 +/- 2.9 mV per decade), and the fully activated current voltage relation showed substantial outward rectification. Selective inhibition of Isus with 50 mumol/L 4AP increased human atrial action potential duration by 66 +/- 11% (P < .01). In conclusion, Isus in human atrial myocytes is due to a very rapidly activating delayed rectifier K+ current, which shows limited slow inactivation, is insensitive to tetraethylammonium, Ba2+, and dendrotoxin, and is highly sensitive to 4AP. These properties resemble the characteristics of channels encoded by the Kv1.5 group of cardiac cDNAs and may represent a physiologically significant manifestation of such channels in human atrium. PMID- 8222079 TI - Differences in cardioprotective efficacy of adrenergic receptor antagonists and Ca2+ channel antagonists in an animal model of dilated cardiomyopathy. Effects on gross morphology, global cardiac function, and twitch force. AB - Turkey poults fed furazolidone (Fz) in high concentrations (700 ppm) develop dilated cardiomyopathy (Fz-DCM). We tested whether five cardioactive agents were cardioprotective in this model of heart failure, ie, whether they prevented dilatation and wall thinning and improved contractile performance. We compared the effects of chronic administration of a beta 1-selective and a nonselective beta-receptor antagonist, an alpha-receptor antagonist, and two Ca2+ channel antagonists in the presence of Fz administration. The greatest cardioprotection was found with treatment with either propranolol or nifedipine. At the gross morphological level, the effect of propranolol (a nonselective beta-adrenergic antagonist) was greater than the effect of atenolol (a selective beta 1 adrenergic antagonist), and the effect of nifedipine was greater than that of verapamil (Ca2+ channel antagonists), with all agents more cardioprotective than phenoxybenzamine (an alpha 1-adrenergic > alpha 2-adrenergic antagonist). Differences in cardioprotective efficacy of each agent increased with increased concentration. These data indicate that the dose and choice of a specific type of Ca2+ channel antagonist or beta-receptor antagonist might be important in the treatment of dilated cardiomyopathy. All agents that were cardioprotective caused similar functional improvements at both the whole heart and isolated muscle levels. Compared with control animals, Fz-DCM animals showed a significant reduction in peak left ventricular (LV) developed pressure (92 +/- 17 versus 143 +/- 24 mm Hg, P < .05), +dP/dt (1151 +/- 219 versus 2454 +/- 549 mm Hg/s), and dP/dt (1128 +/- 291 versus 1875 +/- 396 mm Hg/s), with a significant increase in LV end-diastolic volumes (2.8 +/- 0.7 versus 0.16 +/- 0.1 mL for control animals, P < .05). In contradistinction, LV + dP/dt and -dP/dt values for animals receiving Fz plus a cardioactive agent that demonstrated cardioprotection were not significantly different from control values. Peak LV developed pressures were also similar for Fz animals receiving an agent that demonstrated cardioprotection and control animals not receiving any pharmacologic agent. Isolated muscles from Fz-DCM animals as well as animals receiving Fz plus cardioprotective pharmacologic agents responded normally with regard to increasing extracellular Ca2+ concentrations. Peak twitch forces were greater for animals receiving cardioprotective agents plus Fz than control animals not receiving any pharmacologic agents or Fz alone. At higher stimulation rates, Fz-DCM muscles demonstrated a significantly reduced peak twitch force (4 +/- 0.5 versus 1.5 +/- 0.4 g/mm2 for control muscles versus Fz-DCM muscles, respectively).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8222080 TI - Voltage-dependent Ca2+ channels in resistance arteries from spontaneously hypertensive rats. AB - Alterations in voltage-dependent Ca2+ channels in the arterial smooth muscle cells of spontaneously hypertensive rats (SHR) were investigated using the whole cell voltage clamp and compared with Wistar-Kyoto (WKY) rats. Single cells were freshly isolated from resistance mesenteric arteries from 4- to 5-week-old (young) and 16- to 18-week-old (adult) SHR. Elevated blood pressure was only evident in adult SHR, not in young SHR. In young rats, the Ca2+ channel current density (current amplitude normalized by cell capacitance) was significantly higher (P < .01) in SHR than in WKY rats at the command potential of -10 mV or higher (with 50 mmol/L Ba2+): The current density at 20 mV was -16.8 +/- 1.1 pA/pF in SHR (n = 38 cells) and -11.0 +/- 0.8 pA/pF in WKY rats (n = 30 cells). In adult rats, the difference in current densities disappeared: -15.9 +/- 1.3 pA/pF in SHR (n = 25 cells) and -15.6 +/- 1.5 pA/pF in WKY rats (n = 29 cells). The ratio of maximal amplitude of T-type current to that of L-type current was low in young SHR (0.10 +/- 0.01) compared with the other three groups (0.16 to 0.20). Neither the activation curve nor the steady-state inactivation curve of SHR was different from that of age-matched WKY rats. However, the activation curves in adult rats were shifted to a hyperpolarized direction compared with those of young rats in both strains. These results suggest that the increased activity of voltage-dependent L-type Ca2+ channels of resistance arteries in young SHR may be related to the development of hypertension. The changes observed in adult rats may be due to a secondary modification of the channel during maturation and the presence of hypertension. PMID- 8222081 TI - A redox-based O2 sensor in rat pulmonary vasculature. AB - The effector mechanism of hypoxic pulmonary vasoconstriction (HPV) involves K+ channel inhibition with subsequent membrane depolarization. It remains uncertain how hypoxia modulates K+ channel activity. The similar effects of hypoxia and mitochondrial electron transport chain (ETC) inhibitors on metabolism and vascular tone suggest a common mechanism of action. ETC inhibitors and hypoxia may alter cell redox status by causing an accumulation of electron donors from the Krebs cycle and by decreasing the production of activated O2 species (AOS) by the ETC. We hypothesized that this shift toward a more reduced redox state elicits vasoconstriction by inhibition of K+ channels. Pulmonary artery pressure and AOS, measured simultaneously using enhanced chemiluminescence, were studied in isolated perfused rat lungs during exposure to hypoxia, proximal ETC inhibitors (rotenone and antimycin A), and a distal ETC inhibitor (cyanide). Patch-clamp measurements of whole-cell K+ currents were made on freshly isolated rat pulmonary vascular smooth muscle cells during exposure to hypoxia and ETC inhibitors. Hypoxia, rotenone, and antimycin A decreased lung chemiluminescence ( 62 +/- 12, -46 +/- 7, and -148 +/- 36 counts/0.1 s, respectively) and subsequently increased pulmonary artery pressure (+14 +/- 2, +13 +/- 3, and +21 +/- 3 mm Hg, respectively). These agents reversibly inhibited an outward, ATP independent, K+ current in pulmonary vascular smooth muscle cells. Antimycin A and rotenone abolished subsequent HPV. In contrast, cyanide increased AOS and did not alter K+ currents or inhibit HPV. The initial effect of rotenone, antimycin A, and hypoxia was a change in redox status (evident as a decrease in production of AOS). This was associated with the reversible inhibition of an ATP-independent K+ channel and vasoconstriction. These findings are consistent with the existence of a redox-based O2 sensor in the pulmonary vasculature. PMID- 8222082 TI - Vascular smooth muscle response to ouabain. Relation of tissue Na+ to the contractile response. AB - Smooth muscle responses to Na+ pump inhibition are thought to reflect two elements: a neurogenic contribution, involving catecholamine release from nerve terminals, and a myogenic response, attributed to relations between pump activity, [Na+]i, and [Ca2+]i. In the present study, we describe the time course and magnitude of cell Na+ changes, assessed by two methods, atomic absorption and nuclear magnetic resonance spectroscopy during the myogenic contractile response of rabbit aorta strips to ouabain. A threshold concentration of 3 x 10(-7) mol/L induced a gradual rise in [Na+]i. Both methods showed an essentially identical monotonic rise over 4 to 8 hours from a baseline level of 8 to 10 mmol/L water to a peak, which was approximately fivefold higher. The neurogenic (rapid) and myogenic (delayed and gradual) contractile responses were temporally distinct. Ouabain at 10(-7) mol/L, a concentration 10- to 100-fold lower than the threshold for catecholamine-dependent rapid-onset responses, induced only a delayed and gradual contractile response, which reached a maximum at 6 to 8 hours. With 10( 6) mol/L ouabain, the delayed response of 1.6 +/- 0.2 g peaked at 7.3 +/- 1.1 hours and was sustained for 16 hours. The time course was similar to that for change in [Na+] but somewhat later. Ouabain at 10(-5) and 10(-4) mol/L induced a delayed response that was identical in magnitude but also induced an early rapid contractile response, which was prevented by reserpine or phentolamine pretreatment. These agents did not influence the delayed response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222083 TI - Metabolism and excretion of nitric oxide in humans. An experimental and clinical study. AB - Despite the increasing insight in the clinical importance of nitric oxide (NO), formerly known as endothelium-derived relaxing factor (EDRF), there is limited information about the metabolism and elimination of this mediator in humans. We studied the degradation of NO in healthy subjects inhaling 25 ppm for 60 minutes and in patients with severe heart failure inhaling 20, 40, and 80 ppm in consecutive 10-minute periods. In other healthy subjects, the renal clearance of NO metabolite was measured. The metabolism ex vivo was evaluated by direct incubation of nitrite, the NO oxidation product, in blood from healthy humans. During inhalation of NO, the plasma levels of nitrate increased progressively, both in the healthy subjects (from 26 to 38 mumol/L, P < .001) and in the patients (from 72 to 90 mumol/L, P < .001). Methemoglobin (MetHb) also increased in the healthy subjects (from 7 to 13 mumol/L, P < .001) as well as in the patients (from 19 to 42 mumol/L, P < .01). No change in nitrosohemoglobin (HbNO) was detected, either in the healthy subjects or in the patients. In arterialized blood (O2 saturation, 94% to 99%), incubated nitrite was semiquantitatively converted to nitrate and MetHb. In venous blood (O2 saturation, 36% to 85%) moderate amounts of HbNO were also formed. Plasma and urinary clearance of nitrate in healthy subjects averaged 20 mL/min. We conclude that uptake into the red blood cells with subsequent conversion to nitrate and MetHb is a major metabolic pathway for endogenously formed NO. Nitrate may then enter the plasma to be eliminated via the kidneys.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222084 TI - Regulation of Ca2+ channels by cAMP and cGMP in vascular smooth muscle cells. AB - Whole-cell Ca2+ channel currents in rabbit portal vein cells were recorded using the amphotericin B-perforated patch-clamp technique at 35 degrees C. This technique allowed recording of stable inward currents in the absence of run-down for more than 30 minutes. Depolarizing voltage steps from a holding potential of 70 mV elicited voltage-dependent inward currents. The voltage dependence of inward currents measured in either 2.5 mmol/L Ba(2+)- or 2.5 mmol/L Ca(2+) containing solution were very similar. However, maximum Ba2+ current (obtained at around +10 mV) was approximately 1.5-fold larger than maximum Ca2+ current. Changing the holding potential from -70 to -40 mV decreased inward currents but did not shift the voltage dependence significantly. Inward currents were also completely blocked by the dihydropyridine Ca2+ channel blocker, nicardipine (10 mumol/L), suggesting the presence of predominantly L-type Ca2+ channels in rabbit portal vein cells. Isoproterenol caused small increases in the amplitude of Ba2+ currents in a concentration-dependent manner (10 nmol/L to 1 mumol/L), which were reversed with propranolol. Forskolin (1 mumol/L) or 8-bromo-cAMP (0.1 mmol/L) also caused small increases in the amplitude of Ba2+ currents, suggesting that the stimulatory actions of isoproterenol are importantly linked to the production of cAMP. Higher concentrations of of isoproterenol (10 mumol/L) or forskolin (10 mumol/L) caused a transient increase in Ba2+ currents followed by f decrease in current amplitude. Higher doses of 8-bromo-cAMP (1 mmol/L) and low doses of 8 bromo-cGMP (0.1 mmol/L) inhibited Ba2+ currents, increased the rate of current inactivation, and produced a negative voltage shift in steady-state availability. These results indicate that low concentrations of intracellular cAMP produce modest increases in Ca2+ channel activity, whereas cGMP and higher concentrations of cAMP result in inhibition of Ca2+ channel activity in vascular smooth muscle cells. The observed similarities of cGMP and high concentrations of cAMP on Ba2+ current amplitude, kinetics, and steady-state inactivation suggest mediation by a common mechanism, possibly involving activation of cGMP-dependent protein kinase. PMID- 8222085 TI - Gap junction protein connexin40 is preferentially expressed in vascular endothelium and conductive bundles of rat myocardium and is increased under hypertensive conditions. AB - Gap junction channels consisting of connexin protein mediate electrical coupling between cardiac cells. Expression of two connexins, connexin40 (Cx40) and connexin43 (Cx43), has been studied in ventricular myocytes from normal and hypertensive rats. Polyclonal affinity-purified rabbit antibodies to Cx43 and Cx40 have been used for immunohistochemical analysis on frozen sections from rat heart. These studies revealed coexpression of Cx43 and Cx40 in ventricular myocytes. In addition, Cx40 is preferentially expressed in three distinct regions: first, in the endothelial layer of the heart blood vessels but not in the smooth muscle layer of the arteries; second, in the ventricular conductive myocardium, particularly in the atrioventricular bundle and bundle branches, where Cx43 is not observed; and third, in the myocyte layers close to the ventricular cavities. These results suggest that Cx40 is preferentially expressed in the fast conducting areas of myocardial tissue. Expression of both Cx40 and Cx43 was also found in immunoblots from normal and hypertensive rat myocardiocytes. Under hypertensive conditions (ie, in spontaneous hypertensive rats and in transgenic rats that exhibit hypertension due to expression of an exogenous renin gene), we found a 3.1-fold increase in Cx40 expression, compared with normal myocardium. Furthermore, we detected a 3.3-fold decrease in Cx43 protein level in transgenic hypertensive rats. The coexpression of Cx40 and Cx43 proteins in rat myocytes, their spatial distribution, and the increased amount of Cx40 protein during cardiac hypertrophy suggest that Cx40 may be involved in mediating fast conduction under normal and pathological conditions. The increased expression of Cx40 in hypertrophic heart may be a compensatory mechanism to increase conduction velocity. PMID- 8222086 TI - Determinants of loaded shortening velocity in single cardiac myocytes permeabilized with alpha-hemolysin. AB - Force-velocity relations were obtained from single cardiac myocytes isolated by enzymatic digestion of rat myocardium and permeabilized with the pore-forming staphylococcal toxin alpha-hemolysin. Single cardiac myocytes were attached to a force transducer and piezoelectric translator and viewed with an inverted microscope to allow periodic monitoring of sarcomere length during experiments. Permeabilized cells were activated by immersion in a bath of known [Ca2+]. We report that the Ca2+ sensitivity of cells obtained by enzymatic digestion and permeabilized using alpha-hemolysin is similar to that reported previously for mechanically disrupted ventricular myocardium; however, the tension-pCa relation is less steep in the new preparation. During isotonic measurements, force was clamped to various loads using a rapid-response servo system. All recordings of shortening under load were distinctly curvilinear, and analysis of data involved fitting each shortening recording with a single exponential curve and calculating the value of the slope at the initial time of the load clamp. In addition, the presence of significant resting force at initial sarcomere lengths in these cells required that the possibility of alteration of velocity due to the presence of resting force be addressed. The maximum shortening velocity in fully Ca(2+) activated single ventricular myocytes studied by this method was 2.83 muscle lengths per second on average. The basis for curvilinear shortening is postulated to be multifactorial in cardiac muscle, involving a combination of shortening inactivation and one or more passive elasticities that resist stretch or compression depending on sarcomere length. Shortening velocity shows a dependence on myosin isoform content when cells from a single heart are compared; however, this relation does not hold when cells from different hearts are compared. The behavior of single alpha-hemolysin-permeabilized myocyte shortening under loaded conditions at lower levels of Ca2+ is also described. During submaximal Ca2+ activation, initial shortening velocities are faster than those observed in maximally activated cells. This may be due to contributions of high passive force to increase shortening velocity under conditions of low active force generation, when passive force in the cell is a greater proportion of the total force and there are fewer bound crossbridges. PMID- 8222087 TI - Regulation of hypertrophy and atrophy in cultured adult heart cells. AB - Mechanical loading and alpha-adrenergic receptor stimulation have both been shown to induce hypertrophy in isolated neonatal heart cells. The present study examined the effects of adrenergic hormones and contractile activity on the hypertrophic response in isolated adult feline cardiomyocytes maintained for more than 14 days in insulin- and serum-supplemented medium. Measurements of the hypertrophic response included cell size, total protein content, myosin heavy chain content, and the time course of activation of increased protein synthesis. Reactivation of the "fetal" gene program was evaluated by secretion of atrial natriuretic factor (ANF) into the medium. Significant myocyte hypertrophy was induced in both quiescent myocytes treated with alpha 1-adrenergic agonists and in beating myocytes treated with beta-adrenergic agonists. However, there were both quantitative and qualitative differences in the response to each type of stimulation. alpha-Adrenergic agonists promoted an increase in cell size, protein content, and ANF secretion but not myofibrillar reorganization, which was observed only in beating myocytes. In contrast to results reported for neonatal heart cells, determinants of hypertrophy in beating myocytes exceeded those in nonbeating alpha 1-adrenergic agonist-treated heart cells in every parameter examined. In addition, in the case of both beating and alpha-adrenergic stimulation, there were marked time-dependent variations in rates of protein synthesis over the interval of 4 hours to 7 days of treatment with each type of stimulus. Differences were also encountered in correlations between rates of protein synthesis and protein accumulation over this interval. The effect of beating was particularly important both to the reorganization of myofibrillar structure and the metabolism of myosin heavy chain. In cultures in which beating was inhibited with the calcium channel antagonist nifedipine, the loss of myosin heavy chain was significantly greater than that of total protein. PMID- 8222088 TI - Heat produced by rabbit papillary muscle during anoxia and reoxygenation. AB - Resting heat rate was measured in superfused rabbit papillary muscles at 20 degrees C during 40 minutes of anoxia and subsequent reoxygenation. To reveal the nature of the reactions underlying energy output under such conditions, the data obtained were compared with values predicted from data on chemical change. Before and after the anoxic period, muscles were stimulated at 0.2 Hz, during which time the contraction-related heat rate was measured. During anoxia, muscles were kept at rest or stimulated at 1 Hz. Stimulation was switched off intermittently to determine resting heat rate. Before anoxia, resting heat rate was 8.7 +/- 1.1 (mean +/- SEM) mW.g dry wt-1. During anoxia, it decreased to 38% and 50% of the preanoxic level in resting and stimulated muscles, respectively (P < .05). In resting muscles, heat rate increased with reoxygenation in approximately 10 to 15 minutes to 1.3 times the preanoxic level, whereas this was 3.7 times in stimulated muscles. Resting heat rate returned within 65 (resting muscles) or 150 (stimulated muscles) minutes to the baseline. The ratio of force- and contraction related heat rate, ie, the economy of contraction, was not different before and after anoxia. We estimated that the heat produced by muscles during anoxia was not different from the heat to be expected from the hydrolysis of creatine phosphate, the breakdown of nucleotides, and the formation of lactate. The overshoot in resting heat during reoxygenation of resting muscles could be accounted for by the resynthesis of the energy store. The much larger overshoot in resting heat of stimulated muscles was due to the contracture. The finding that the economy of contraction was not altered by anoxia and reoxygenation suggests that both sarcoplasmic reticulum Ca(2+)-ATPase and myofibrillar ATPase are depressed by anoxia and that the enhancement of cytosolic calcium transients with reoxygenation, reported in other studies on papillary muscle, results from reduced binding of calcium rather than from enhanced release. PMID- 8222089 TI - Intraluminal flow-initiated hyperpolarization and depolarization shift the membrane potential of arterial smooth muscle toward an intermediate level. AB - We examined the effect of intraluminal flow of physiological saline on the membrane potential of vascular smooth muscle cells in isolated rabbit cerebral arteries. Intraluminal flow (20 microL/min) caused a depolarization of 4.8 +/- 0.7 mV in muscle cells with a resting membrane potential of -62.5 +/- 1.2 mV (n = 19). However, when cells were depolarized to -48.7 +/- 1.8 mV using histamine and serotonin, the response to intraluminal flow was the opposite, a hyperpolarization of 5.6 +/- 1.0 mV (n = 9). These opposing effects of flow on membrane potential appear to balance at -57.8 +/- 1.1 mV (n = 31). Our results suggest that intraluminal flow may affect the level of basal tone present in arteries in vivo through modulating the membrane potential of vascular smooth muscle cells by concurrently activated depolarization and hyperpolarization. PMID- 8222090 TI - Role of nitric oxide, muscarinic receptors, and the ATP-sensitive K+ channel in mediating the effects of acetylcholine to mimic preconditioning in dogs. AB - The aims of the current study were to examine the efficacy of acetylcholine (ACh) to mimic ischemic preconditioning in dogs and to determine the role of nitric oxide (NO), muscarinic receptors, and ATP-sensitive K+ (KATP) channels in mediating its effects. Barbital-anesthetized open-chest dogs were subjected to 60 minutes of left anterior descending coronary artery (LAD) occlusion followed by 4 hours of reperfusion. Preconditioning was elicited by 10 minutes of LAD occlusion followed by 10 minutes of reperfusion before the 60-minute occlusion period. ACh (3 or 10 micrograms/min) or an equivalent volume of saline was infused into the LAD for 10 minutes, followed by a 10-minute drug-free period before the 60-minute ischemic insult. In other groups, the specific NO synthesis inhibitor NG monomethyl-L-arginine (L-NMMA, 4 mg/min), the muscarinic receptor antagonist-NO synthesis inhibitor nitro-L-arginine methyl ester (L-NAME, 3 mg/min), or the specific KATP channel blocker 5-hydroxydecanoate (5-HD, 3 mg/min) was infused with ACh into the LAD for 10 minutes. The infusion of L-NAME, L-NMMA, or 5-HD was started 2 minutes before ACh infusion. Transmural myocardial blood flow was measured at 5 minutes of occlusion, and infarct size was determined by triphenyltetrazolium staining and expressed as a percentage of the area at risk. There were no significant differences in collateral blood flow or the area at risk between groups. Preconditioning produced a marked reduction (P < .05) in infarct size (6.2 +/- 3.0% versus 26.1 +/- 5.7% in the control group).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222091 TI - Efficient gene transfer into myocardium by direct injection of adenovirus vectors. AB - Previous studies have established that gene transfer into myocardial cells in vivo is detectable after direct injection of plasmid DNA. Recently, adenovirus vectors have been shown to provide an efficient method for gene transfer into a wide range of tissues. Therefore, this study sought to assess the efficiency and stability of adenovirus-mediated gene transfer into myocardium and to compare this method with that using plasmid-based gene transfer techniques. Adult rats underwent myocardial injection via a subdiaphragmatic approach. Gene transfer efficiency was compared using direct injection of an adenovirus vector encoding for the marker gene beta-galactosidase (beta-gal), a control adenovirus vector encoding for the cystic fibrosis transmembrane conductance regulator gene, a plasmid encoding for beta-gal, or a control plasmid. Hearts infected with an adenovirus vector containing the beta-gal gene showed significantly increased beta-gal enzymatic activity compared with hearts injected with beta-gal plasmid. Histological examination revealed that cardiac myocytes were the target of adenovirus-mediated gene transfer. A time course of gene expression showed that beta-gal enzymatic activity peaked during the first week following injection. Adenovirus vectors provide an efficient but transient method for in vivo gene expression in myocardium. PMID- 8222092 TI - Stereoselective actions of thiadiazinones on canine cardiac myocytes and myofilaments. AB - Thiadiazinones are cardiotonic agents that have potent, direct, and stereoselective actions on the myofilament response to Ca2+ in intact myocardium. Their mechanism of action is unknown. We studied the effects of racemic thiadiazinone, EMD 53998 (5-[1-(3,4-dimethoxybenzoyl)-1,2,3,4-tetrahydro-6 quinolyl]-6-meth yl-3,6- dihydro-2H-1,3,4-thiadiazin-2-one), and its enantiomers on Ca2+ signaling in myocytes, myofilaments, and myofilament proteins. Intact canine ventricular myocytes responded to the positive enantiomer, EMD 57033, with an increase in the extent of shortening during twitch contractions without increasing the peak amplitude of the Ca2+ transient. The negative enantiomer, EMD 57439, also increased the extent of shortening, but in this case there was a concentration-dependent increase in the peak amplitude of the Ca2+ transient. This is predicted from in vitro data showing that this enantiomer is a relatively potent inhibitor of phosphodiesterase activity. There was no effect of EMD 57439 on the relation between pCa and actomyosin Mg-ATPase activity of canine heart myofibrils. In contrast, EMD 57033 shifted the pCa-Mg-ATPase activity relation to the left. There was no effect of either enantiomer on Ca2+ binding to myofilament troponin C. Moreover EMD 57033, but not EMD 57439, stimulated actomyosin ATPase activity of myofilament preparations in which either troponin or troponin tropomyosin had been extracted. EMD 57033 had no effect on Mg-ATPase activity of pure ventricular myosin. EMD 57033 also stimulated the velocity of actin filament sliding on myosin heads adhered to nitrocellulose-coated glass coverslips. We propose that the action of EMD 57033 is at the actin-myosin interface on a "receptor" that may be on actin or the crossbridge. Drug binding to this domain appears to reverse the inhibition of actin-myosin interactions by troponin tropomyosin and also to promote transition of crossbridges from weak to strong force-generating states. PMID- 8222093 TI - Recombinant hirudin in patients with chronic, stable coronary artery disease. Safety, half-life, and effect on coagulation parameters. AB - BACKGROUND: Because the specific antithrombin hirudin prevents platelet-rich arterial thrombus and accelerates thrombolysis in a variety of animal models, it has promise as antithrombotic therapy. We therefore studied the half-life, effect on anticoagulant parameters, and safety of hirudin in patients with coronary artery disease. METHODS AND RESULTS: Thirty-eight men and 1 woman (age [mean +/- SD], 60.4 +/- 6.9 years) with angiographic coronary disease were allocated in a single-blind ascending dosage study to a 6-hour i.v. infusion of recombinant hirudin (CGP 39,393) or matching placebo. The median terminal half-life for hirudin, measured by ELISA, was 2.7, 2.3, 2.9, 3.1, and 2.0 hours for the 0.02, 0.05, 0.1, 0.2, and 0.3 mg.kg-1 x h-1 groups, respectively. Activated partial thromboplastin times (aPTT) at 3, 4, and 6 hours were averaged into a plateau value. The aPTT plateau-to-baseline ratios were 1.5 +/- 0.1, 2.0 +/- 0.1, 2.3 +/- 0.1, 2.7 +/- 0.1, and 2.9 +/- 0.1, respectively, with hirudin infused at 0.02, 0.05, 0.1, 0.2, and 0.3 mg.kg-1 x h-1. From 62% to 77% of the aPTT plateau value was seen within 30 minutes of starting the infusions and was directly related to dose. The aPTT-to-baseline ratios correlated well with plasma hirudin levels (r = .88), whereas poor correlation and sensitivity were observed between plasma hirudin levels and activated coagulation time (ACT)-to-baseline ratios (r = .44). Plasma levels of hirudin and ACT in seconds correlated overall well (r = .80), but considerable overlap occurred between baseline ACT and ACT at plasma hirudin concentrations < 1000 ng/mL. Prothrombin times were significantly prolonged only at a dosage of > or = 0.05 mg.kg-1 x h-1 and were 11.8 +/- 0.5 (INR = 1.0), 12.3 +/- 0.7 (INR = 1.1), 13.3 +/- 1.2 (INR = 1.4), 14.2 +/- 0.4 (INR = 1.7), and 15.8 +/- 0.9 (INR = 2.3) seconds for each respective hirudin dosage. Thrombin times were beyond range (> 600 seconds) at 6 hours in all except 2 patients who received the lowest dosage. All parameters returned to baseline between 8 and 18 hours after the infusion. Bleeding times were not significantly prolonged. No side effects occurred. No antibodies to hirudin were detected 2 weeks after the infusion. CONCLUSIONS: Recombinant hirudin has a terminal half-life of 2 to 3 hours. The aPTT correlates well with plasma levels of hirudin and allows close titration over a wide range of anticoagulation, while ACT and prothrombin time are relatively insensitive for monitoring hirudin administration. At anticoagulant levels effective in experimental thrombosis, a 6-hour infusion of hirudin is well tolerated and safe in a predominantly male group of patients with stable coronary atherosclerosis. PMID- 8222094 TI - Evaluation of bioprosthetic valve-associated thrombus in ventricular assist device patients. AB - BACKGROUND: Thromboembolic events may be related to thrombotic deposition on prosthetic valves. In a left ventricular assist device (LVAD) that contains two porcine pericardial bioprosthetic valves in addition to significant associated biomaterial placement, this may be particularly true. Thrombotic deposits on valves removed from LVADs at autopsy or heart transplantation were scored to determine (1) the nature and location of valvular deposition, (2) whether deposition was related to thromboembolic events, (3) correlations between deposition and patient hemodynamic and coagulation parameters, and (4) implant time dependency. METHODS AND RESULTS: Novacor LVADs were implanted in 23 patients as a bridge to transplantation for 1 to 303 days. Photographs of the concave (downstream) and convex (upstream) side of the inflow and outflow valve were made at explant and later scored for (1) total thrombus area (10 = equivalent of cusp area), (2) percent of cusp area occupied by solid thrombus, (3) thrombus color (10 = dark red, 0 = white), and (4) average percent of valve strut height involved with thrombus (from a side view). The inflow valve was shown to have heavier and redder deposition than the outflow valve. This was also true for the concave versus the convex side. Heaviest deposition was seen on the inflow valve concave side, which rests within the LVAD pumping sac and may be subject to poor convection. Patients with neurological thromboembolic events (8/23) during implantation had heavier deposition on the inflow valve concave side (5.7 +/- 2.7 versus 4.6 +/- 2.2, P < .05). Pump volumetric output was also found to negatively correlate with thrombus area on this valve and side (r = -.61, P = .002). Platelet release (platelet factor 4) was correlated with thrombus involvement on the upstream (convex) side of the inflow valve (r = .82, P = .002). No significant dependence of deposition on the implant time was found. CONCLUSIONS: Valve thrombus deposition was related to thromboembolic events. Pump volumetric output and platelet release were found to be related to deposition. These results may have implications for the role of hemodynamics and platelet activation in thromboembolism associated with prosthetic valve placement in general. PMID- 8222095 TI - Predictive value of tissue plasminogen activator mass concentration on long-term mortality in patients with coronary artery disease. A 7-year follow-up. AB - BACKGROUND: The fibrinolytic system is part of the defense against thrombotic and cardiovascular events, but so far no study has shown that clinical measurements of fibrinolytic key components such as tissue plasminogen activator (t-PA) or plasminogen activator inhibitor type 1 (PAI-1) have any predictive value beyond 3 years. METHODS AND RESULTS: In 1983 through 1985, 213 consecutive patients with angina pectoris and angiographically verified coronary artery disease were sampled, and the mass concentration of t-PA and the activity of PAI-1 were measured in citrated plasma samples. At a mean follow-up time of 7 years, the all cause mortality was checked. No patient was lost to follow-up. The data were analyzed by Cox regression, and t-PA mass concentration was found to be the only laboratory risk factor significantly related to mortality in all patients (P < .022) and also in the major subgroup (78% of all patients) subjected to coronary bypass surgery (P < .027). In the latter subgroup, body mass index was also related to mortality. CONCLUSIONS: An increased mass concentration of t-PA is a new risk factor of long-term mortality in patients with angina pectoris and coronary artery stenosis. This paradoxical effect probably reflects increased t PA levels attributable to enzyme inhibitor complex formation in subjects with increased plasma levels of t-PA inhibitors. PMID- 8222096 TI - Profound platelet degranulation is an important side effect of some types of contrast media used in interventional cardiology. AB - BACKGROUND: Thrombotic complications occurring during coronary angiography and percutaneous transluminal coronary angioplasty (PTCA) are relatively frequent and can be influenced by the type of radiographic contrast media used. Low osmolar contrast media (LOCM), both ionic and nonionic, have been considered to be safer than the older high osmolar contrast media (HOCM), causing less haemodynamic and symptomatic side effects. Recently, however, nonionic LOCM have been associated with an increased incidence of thrombotic events, including coronary occlusion and stroke. METHODS AND RESULTS: The effects of commonly used contrast media on platelets in native blood were investigated using immunolabeling and flow cytometry to detect platelet activation in vitro. A nonionic LOCM (Omnipaque) caused profound platelet degranulation in nearly 80% of platelets compared with 2 to 3% of platelets in the control. Conversely, an ionic HOCM (Urografin) caused only 25% degranulation, whereas an ionic LOCM (Hexabrix) caused no platelet activation and, furthermore, it inhibited the effects of thrombin on platelets. Platelet degranulation, quantified by immunolabeling, was paralleled by release of beta-thromboglobulin and platelet factor 4 from platelet alpha-granules. Blood from patients anticoagulated with heparin and pretreated with standard-dose aspirin in preparation for PTCA showed the same pattern of contrast media-induced platelet activation as normal subjects. CONCLUSIONS: These results suggest that the type of contrast media used during invasive imaging of the vasculature could have a significant effect on platelets. Platelet degranulation within a PTCA damaged vessel would be increased by a nonionic contrast medium, releasing procoagulant molecules and platelet-derived growth factors into the damaged vessel lumen, which might contribute to acute thrombosis and the initiation of the restenosis process. PMID- 8222097 TI - Aspirin versus heparin to prevent myocardial infarction during the acute phase of unstable angina. AB - BACKGROUND: Antiplatelet therapy with aspirin and antithrombotic therapy with heparin both prevent the complications of unstable angina; however, no definitive data exist on the relative clinical efficacy of the two drugs. METHODS AND RESULTS: Aspirin (325 mg bid) or heparin (5000-U intravenous bolus followed by a perfusion titrated to the APTT) were compared in a double-blind randomized trial of 484 patients in two cohorts enrolled sequentially. The study was initiated at admission to hospital at a mean of 8.3 +/- 7.8 hours after the last episode of pain. End points were assessed 5.7 +/- 3.3 days later, when the decision for long term management was made. Myocardial infarction occurred in 2 (0.8%) of the 240 patients randomized to heparin and in 9 (3.7%) of the 244 randomized to aspirin (P = .035), an odds ratio of 0.22 and a risk difference of 2.9% (95% confidence limits, 0.3% to 5.6%) with heparin. The only death resulted from a myocardial infarction in an aspirin patient. Survival curves with Cox logistic regression analysis showed that the improvement in survival without myocardial infarction with heparin (P = .035) was independent of other baseline characteristics. CONCLUSIONS: This study documents that heparin prevents myocardial infarction better than aspirin during the acute phase of unstable angina. PMID- 8222098 TI - Excimer laser coronary angioplasty of aorto-ostial stenoses. Results of the excimer laser coronary angioplasty (ELCA) registry in the first 200 patients. AB - BACKGROUND: Percutaneous transluminal coronary angioplasty (PTCA) of aorto-ostial stenosis has been associated with a lower rate of acute success, a high risk of vessel closure, and late restenosis. The purpose of this report is to document a prospective multicenter trial of excimer laser coronary angioplasty (ELCA) of aorto-ostial stenosis involving the coronary arteries and saphenous vein grafts. METHODS AND RESULTS: Between December 1989 and May 1992, 206 aorto-ostial ELCA procedures were performed on 209 stenoses in 200 patients. Canadian Cardiovascular Society class III or IV angina was present in 76%. The distribution of stenosis locations was left main coronary (LM) in 26 (12%), right coronary (RCA) in 124 (59%), and vein grafts (VG) in 59 (28%). Adjunctive PTCA was performed in 72%. Procedure success defined as < or = 50% diameter stenosis without major complications was achieved in 90% (LM, 92%; RCA, 89%, VG, 90%). Quantitative angiographic analysis documented an improvement in stenosis diameter from 0.8 +/- 0.5 mm or 76 +/- 14% at baseline to 2.1 +/- 0.6 mm or 36 +/- 15% at completion (P < .01). The majority of the acute gain in diameter (1.0 +/- 0.6 mm) resulted from ELCA. A major complication during hospitalization occurred in 3.9% (death, 0%; Q-wave myocardial infarction, 0.5%; bypass surgery, 3.4%). The only logistic regression univariate and multivariate predictor of procedure failure was female gender. Six-month angiographic follow-up, available in 51% of eligible patients, documented an average lumen diameter of 1.7 +/- 1.0 mm and mean diameter stenosis of 46 +/- 26%. Restenosis (> 50% diameter stenosis) occurred in 39% (LM, 64%; RCA, 35%; VG, 35%). Restenosis was less likely when residual stenosis was < or = 35% (28% versus 53%, P < .05). Clinical events at follow-up were death, 2.7%; bypass surgery, 6.5%; myocardial infarction, 2.2%; and repeat angioplasty, 16.2%. Of the remainder, 78% were asymptomatic, class I or II for anginal symptoms. An adverse event during follow-up was more than twice as likely in the group with LM (50.0% versus 21.1%, P < .02). CONCLUSIONS: ELCA is acutely effective and safe therapy in patients with aorto-ostial stenosis. Six-month restenosis, adverse-event rates were higher and functional status was poorer in the group with LM stenosis. ELCA may be considered as an alternative to bypass surgery in carefully selected patients with isolated aorto-ostial stenosis of the RCA and saphenous vein grafts. PMID- 8222099 TI - Safety and efficacy of recombinant hirudin (CGP 39 393) versus heparin in patients with stable angina undergoing coronary angioplasty. AB - BACKGROUND: Enhanced thrombin activity has been associated with acute and long term complications following balloon angioplasty (percutaneous transluminal coronary angioplasty (PTCA). We evaluated, in a 2-to-1 randomized, double-blind trial, the effects of recombinant hirudin, CGP 39 393, relative to unfractionated sodium heparin on periprocedural events, bleeding, early angiographic outcome, and coagulation in 113 patients with stable angina undergoing PTCA. METHODS AND RESULTS: Prior to PTCA, 20 mg CGP 39 393 was administered as a bolus, followed by continuous infusion at a rate of 0.16 mg.kg-1 x h-1, or 10,000 IU sodium heparin was administered as a bolus and continued at a rate of 12 IU.kg-1 x h-1 for 24 hours. Infusion was adjusted to activated partial thromboplastin time (APTT) levels. ST segment was monitored for 24 hours, and angiograms were analyzed with quantitative technique (QCA). In 74 CGP 39 393- and 39 heparin-treated patients, 132 lesions were dilated. Myocardial infarction and/or emergency coronary bypass surgery occurred in 1 (1.4%) CGP 39 393 patient compared with 4 (10.3%) heparin patients (relative risk, 7.6; 95% confidence interval, 0.9, 65.6). At 24 hours, complete perfusion was present in 91% heparin and 100% CGP 39 393 patients. Significant ST segment displacement was found in 11% of heparin versus 4% of CGP 39 393 subjects. Bleeding occurred only at the puncture site in 4 CGP 39 393 treated patients. QCA did not reveal significant differences between the groups. APTT values were more often in the target range and more stable in CGP 39 393 patients. Levels of thrombin-antithrombin III complexes, prothrombin fragment F1+2, and fibrinopeptide A indicated that CGP 39 393 was an effective inhibitor of thrombin activity. CONCLUSIONS: CGP 39 393 can safely be administered to patients undergoing elective PTCA for stable anginal symptoms and may have a more favorable anticoagulant profile than heparin. PMID- 8222100 TI - Use of direct angioplasty for treatment of patients with acute myocardial infarction in hospitals with and without on-site cardiac surgery. The Myocardial Infarction, Triage, and Intervention Project Investigators. AB - BACKGROUND: In the Myocardial Infarction, Triage, and Intervention (MITI) registry of acute myocardial infarction, 441 (12%) of 3750 patients had direct angioplasty as initial treatment. Approximately half (233) were performed in hospitals with no on-site surgery. METHODS AND RESULTS: Procedure success rates, use of emergent surgery, and factors influencing outcome were compared in both angioplasty groups as well as with 653 patients treated with thrombolytic therapy in the same hospitals. There was no difference in baseline characteristics between patient groups treated by angioplasty in the two types of hospitals. Patency was established in 88% of patients. Only 1.4% underwent emergent surgery. Overall, survival was 93% but was significantly worse after a failed procedure in all ECG and hemodynamic subsets as well as in those with prior bypass surgery. In a multivariate analysis, age, initial heart rate, blood pressure, and prior bypass surgery but not type of hospital were predictive of survival. Survival rates were similar, but there tended to be fewer strokes (0.6% versus 2.1%, P = .12), shorter hospital stays (7.0 versus 8.1 days, P < .001), and less recurrent ischemia (20% versus 30%, P = .009) in patients treated by angioplasty compared with thrombolysis. Readmission and reinfarction rates were similar for both treatments. CONCLUSIONS: Observations from this community registry suggest that mortality after direct angioplasty is low and the use of emergent surgery is infrequent. Outcome in this registry study was dependent on initial hemodynamic findings and infarct location but not on the presence of on-site surgery. Compared with thrombolytic therapy, the incidence of complications was the same or lower, but this needs confirmation in randomized trials. PMID- 8222101 TI - Response of human coronary arteries to serotonin after injury by coronary angioplasty. AB - BACKGROUND: Atherosclerotic stenoses that have exaggerated vasomotor responses are especially prone to restenosis after coronary angioplasty. Experimental studies show that vasomotor responses in normal vessels are altered by acute mechanical injury, an alteration that in part reflects changes in the functional characteristics of endothelium that has regenerated after injury. METHODS AND RESULTS: We examined, by quantitative coronary arteriography, the response of dilated and control coronary segments to intracoronary infusions of graded doses of serotonin, an endothelium-dependent vasoactive agent, and to intracoronary injection of isosorbide dinitrate, an endothelium-independent smooth muscle dilator in 15 patients who had undergone a single percutaneous transluminal coronary angioplasty procedure and who had no clinical features of variant angina. Dose-dependent constriction to serotonin occurred at all measured sites. The mean +/- SEM diameter reductions, expressed as percent reduction in baseline diameter that was observed at proximal (18.1 +/- 2.9, 18.8 +/- 2.2) and distal (30.9 +/- 4.4, 35.4 +/- 5.3) control sites in the dilated and nondilated vessels, respectively, at the highest dose, were similar. The degree of constriction in distal segments was significantly (P < .01) greater than that in proximal segments. Total or subtotal occlusion occurred at the angioplasty site in 4 patients at the highest infused dose (10(-4) mol/L). The mean percent reduction in baseline diameter at previously dilated sites (53.8 +/- 5.9) at this dose was significantly (P < .05) greater than that observed at the adjacent proximal control sites and similar to that observed at distal control sites. All segments dilated significantly after intracoronary injection of isosorbide dinitrate. CONCLUSIONS: In dilated and nondilated vessels, serotonin caused significantly more marked constriction in distal than in proximal vessel segments. In dilated vessels, the vessel segments that had been subjected to angioplasty had a constrictor response to serotonin that was more marked than at adjacent proximal control sites and equivalent to that in the distal vessel segments. This enhanced constrictor response could be related to changes in endothelial cell function after regeneration or to hyperreactivity of smooth muscle cells at the angioplasty site. PMID- 8222102 TI - Clinical and angiographic outcomes after coronary artery stenting for acute or threatened closure after percutaneous transluminal coronary angioplasty. Initial results with a balloon-expandable, stainless steel design. AB - BACKGROUND: Acute occlusion after balloon coronary angioplasty is associated with an increased risk of angina, emergency coronary artery bypass grafting (CABG), myocardial infarction (MI), and death. Stents offer a way of restoring patency and avoiding these complications. METHODS AND RESULTS: One hundred sixteen patients underwent attempted stent placement for imminent or total acute closure after PTCA. In 103 patients (110 stents, 105 procedures) the stent was successfully deployed (89%). Angiographic success (final diameter stenosis of < 50%) was achieved in 94 placements (85%). Seventy-one phase 2 procedures (CABG optional, n = 96; phase 1, CABG required, n = 9) were angiographically successful without complications of death, Q-wave myocardial infarction, or CABG (clinical success 74%). Stent placement was associated with resolution of ST-segment deviation and angina in 84% of patients. Five deaths and 5 Q-wave MIs occurred during hospitalization. Two deaths were related to pulmonary insufficiency from chronic lung disease and one patient died after rescue stent placement for left main coronary artery occlusion during routine angiography. Another patient died after CABG was followed by right ventricular MI. The last death occurred in an elderly patient who suffered a stroke while on intravenous heparin. During hospitalization nine patients developed reocclusion after stent placement (8.6% of procedures) and six had repeat PTCA. CABG was performed after 29 stent procedures (28%). The first nine patients underwent CABG as a mandate of the phase 1 protocol. In addition, nine patients had CABG after stenting with a good angiographic result but with a large amount of myocardium at risk. Clinical follow-up was obtained in all patients at a median of 14 months (range, 2 to 43). There were three late deaths (3%), two Q-wave myocardial infarctions (2%), 16 repeat PTCAs (16%), and 15 CABG procedures (15%). Angiographic restenosis (percent diameter > or = 50%) using caliper measurements was found in 30 of 57 patients (53%) at a median of 4 months (93% of patients eligible). A total of 41 procedures were successful and unaccompanied by death, emergency or elective coronary artery bypass grafting, or angiographic restenosis in follow-up. Restenosis and/or clinical events (death, MI, CABG, repeat PTCA) were associated with non-Q MI, hypertension, diabetes, left circumflex coronary artery stenting, saphenous vein graft stenting, smaller caliber artery stenting, higher balloon to artery ratios, and shorter inflation times. CONCLUSIONS: Coronary artery stenting for acute closure after PTCA relieves myocardial ischemia and provides an alternate means of treatment. This series includes early learning curve experience; 70% (67 of 96) of patients were spared emergency coronary artery bypass graft surgery when this adverse outcome occurred. Certain clinical and angiographic subsets are at increased risk for restenosis and future cardiac events. PMID- 8222103 TI - After correcting for worse baseline characteristics, women treated with thrombolytic therapy for acute myocardial infarction have the same mortality and morbidity as men except for a higher incidence of hemorrhagic stroke. The Investigators of the International Tissue Plasminogen Activator/Streptokinase Mortality Study. AB - BACKGROUND: In the prethrombolytic era, women with myocardial infarction were reported to have a worse outcome than men. This analysis evaluates the association of sex with morbidity and mortality after thrombolytic therapy. METHODS AND RESULTS: Data were analyzed from 8261 of the 8387 randomized patients with acute myocardial infarction who received thrombolytic therapy in the International Tissue Plasminogen Activator/Streptokinase Mortality Study (baseline data were missing for 126 patients) and were followed for 6 months. Women made up 23% (n = 1944) of the study population. Baseline characteristics were worse in women: they were 6 years older, were more likely to have a history of previous infarction (P < .01), antecedent angina (P < .01), hypertension (P < .0001), or diabetes (P < .0001); were in a higher Killip class on admission (P < .0002); and received thrombolytic therapy 18 minutes later than men (P < .0001). Fewer women were smokers (P < .0001). Women had a higher hospital (12.1% versus 7.2%, P < .0001) and 6-month mortality (16.6% versus 10.4%, P < .0001) and were more likely to develop cardiogenic shock (9.1% versus 6.3%, P < .0001), bleeding (7.2% versus 5.3%, P < .01), and hemorrhagic (1% versus 0.3%, P < .001) or total stroke (2.2% versus 1.1%, P < .0001) during hospitalization. Reinfarction rates and requirement for angioplasty or surgery did not differ. After correction for worse baseline characteristics, women had similar morbidity and mortality apart from a significantly higher incidence of hemorrhagic stroke, which remained significant even after accounting for weight and treatment allocation (odds ratio, 2.90; P < .01). CONCLUSIONS: After thrombolytic therapy for acute myocardial infarction, women have similar morbidity and mortality to men but suffer from a higher incidence of hemorrhagic stroke. PMID- 8222104 TI - Differences between men and women in hospital mortality associated with coronary artery bypass graft surgery. The Northern New England Cardiovascular Disease Study Group. AB - BACKGROUND: A prospective study of patients undergoing coronary artery bypass graft surgery (CABG) was conducted to examine differences in hospital mortality by sex. Outcome data on 3055 CABG patients undergoing operation between 1987 and 1989 were examined for differences in patient, disease, and treatment factors. METHODS AND RESULTS: Odds ratios (OR), risk differences, and 95% confidence intervals (CI95%) were calculated. Mortality rates for women (7.1%) and men (3.3%) differed, the OR (women versus men) being 2.23 (CI95%, 1.58 to 3.15). Women were older, more often diabetic, and had more urgent or emergent surgery; adjustment yielded an OR (women versus men) of 1.75 (CI95%, 1.17 to 2.63). Body surface area (BSA) was associated with risk of death in both sexes (P = .007) and positively associated with coronary artery luminal diameters. After adjustment for BSA, sex was no longer significantly associated with mortality (OR [women versus men] of 1.18; CI95%, 0.72 to 1.95). Internal mammary artery (IMA) grafting was performed less frequently among women than men (64.8% versus 78.4%, P < .001). Smaller BSA and absence of IMA grafting were each associated with increased risk of death (RD) from heart failure. Risk of death from heart failure (RD [women minus men] = 2.05; CI95%, 0.89 to 3.22) and hemorrhage (RD [women minus men] = 0.63; CI95%, 0.13 to 1.13) was greater among women; these accounted for 71.1% of the sex-specific difference in mortality rates. CONCLUSIONS: Excess risk of hospital mortality among women having CABG was largely the consequence of death from heart failure and, to a lesser extent, from hemorrhage. Smaller BSA (probably because of its association with coronary artery luminal diameter) and the absence of IMA grafting were each associated with increased risk of death from heart failure. PMID- 8222105 TI - Thromboxane A2 and prostacyclin biosynthesis in children and adolescents with pulmonary vascular disease. AB - BACKGROUND: The pathogenesis of pulmonary vascular disease in children with congenital heart disease is incompletely understood. Thromboxane (TX) A2 and prostacyclin (PGI2) have opposing effects on platelet aggregation and pulmonary vascular smooth muscle. An imbalance in their biosynthesis could contribute to the progressive increase in pulmonary vascular resistance seen in older untreated patients with pulmonary hypertensive congenital heart disease and the thrombotic complications they may develop. METHODS AND RESULTS: We investigated TXA2 and PGI2 biosynthesis in 15 young children (0.2 to 2.25 years old) with congenital heart disease with increased pulmonary blood flow and potentially reversible pulmonary vascular disease by measuring urinary excretion of 2,3-dinor-TXB2 and 2,3-dinor-6-oxoprostaglandin (PG) F1 alpha and compared the findings with those in 16 healthy children (0.5 to 2.8 years old). 2,3-Dinor-TXB2 excretion was greater in the patients than in control subjects (1253 +/- 161 versus 592 +/- 122 ng/g creatinine; P < .001). Excretion of 2,3-dinor-6-oxo-PGF1 alpha was 452 +/- 54 compared with 589 +/- 95 ng/g creatinine in control subjects. In 5 patients who underwent successful cardiac surgery > or = 1 year later excretion of 2,3 dinor-TXB2 decreased from 1100 +/- 298 to 609 +/- 131 ng/g creatinine (P < .05), a value comparable to those in 5 healthy children of similar age (749 +/- 226 ng/g creatinine). We also compared 15 patients (11 to 23 years old) with advanced irreversible pulmonary vascular disease with 19 healthy control subjects (10 to 23 years old). The ratio of TX to PGI2 metabolite excretion was greater in the patients than in control subjects (3.5 +/- 0.6 versus 2.0 +/- 0.3; P < .05). CONCLUSIONS: There is increased 2,3-dinor-TXB2 excretion in children with congenital heart disease and a high pulmonary blood flow that may reflect an imbalance in biosynthesis of TXA2 and PGI2 in the pulmonary vascular bed. The imbalance may contribute to the progressive development of increased pulmonary vascular resistance and persists in older patients whose heart defects are uncorrected. PMID- 8222106 TI - Intracoronary albunex. Its effects on left ventricular hemodynamics, function, and coronary sinus flow in humans. AB - BACKGROUND: Albunex is a recently developed ultrasonic contrast agent made from sonicated human serum albumin. The effects on left ventricular hemodynamics, function, and coronary sinus flow of intracoronary Albunex in humans have not been reported. METHODS AND RESULTS: Eighteen patients with known or suspected coronary artery disease were examined at the time of coronary arteriography with simultaneous two-dimensional echocardiography and left ventricular catheter-tip manometry. Intracoronary injections of Albunex into the left main coronary artery were performed, as were injections of 5% human serum albumin and iohexol, a widely used angiographic contrast agent. Mean coronary sinus flow was determined before and after injections of iohexol and 2 mL of Albunex. Injection of 1 mL of Albunex induced no changes in any of the measured hemodynamic parameters (heart rate, peak left ventricular [LV] systolic pressure, LV end-diastolic pressure, positive or negative LV dP/dt, or time constant of relaxation) or echocardiographic determinants of LV function (regional wall motion and global ejection fraction). Injection of 2 mL or more of Albunex caused small, transient (less than 30 seconds) changes in measures of isovolumic relaxation (negative LV dP/dt; 95% confidence interval: mean, -2.41 [-4.3, -0.52] and tau 1; confidence interval mean, 3.52; [1.48, 5.58]) but not in functional measures. Intracoronary injection of 5% human serum albumin had no effect. Iohexol induced small but significant changes in both systolic and diastolic parameters, which lasted beyond 30 seconds after injection. Mean coronary sinus blood flow increased. CONCLUSIONS: The effects of Albunex on hemodynamics, left ventricular function, and coronary sinus blood flow compare favorably with iohexol. Albunex can be considered to be an essentially inert contrast agent if used in patients with stable coronary artery disease. PMID- 8222107 TI - Use of inhaled nitric oxide and acetylcholine in the evaluation of pulmonary hypertension and endothelial function after cardiopulmonary bypass. AB - BACKGROUND: Increased pulmonary vascular resistance is common in congenital heart disease and is exacerbated by cardiopulmonary bypass (CPB). We investigated whether CPB is responsible for pulmonary endothelial dysfunction and contributes to postoperative pulmonary hypertension. METHODS AND RESULTS: We infused the endothelium-dependent vasodilator acetylcholine (ACH) into the pulmonary circulation of pulmonary hypertensive children with congenital heart disease either before (n = 12) or after (n = 22) surgical repair on CPB. The dose response to ACH (10(-9) to 10(-6) M) was recorded for all hemodynamic variables. Nine additional postoperative patients were studied with ACH followed by inhalation of 80 ppm nitric oxide, an endothelium-independent smooth muscle relaxant. Plasma levels of cyclic GMP (cGMP) were measured before and after ACH and nitric oxide administration. Pulmonary vasodilation with 10(-6) M ACH was seen in all preoperative patients but was markedly attenuated in postoperative patients. Baseline pulmonary vascular resistance (5.6 +/- 1.0 U x m2) fell 46 +/- 5% in preoperative patients but declined only 11 +/- 4% from baseline (5.8 +/- 0.9 U x m2) in postoperative patients (P < .002). However, inhalation of 80 ppm nitric oxide after ACH infusion in postoperative patients lowered pulmonary vascular resistance by 33 +/- 4% (P < .0002 compared with postoperative ACH response) with minimal effects on the systemic circulation. This finding suggests that the capacity for smooth muscle relaxation and pulmonary vasodilation was present in postoperative patients but could not be induced by ACH. Plasma levels of cGMP in postoperative patients were unchanged after acetylcholine infusion but rose more than threefold during pulmonary vasodilation with nitric oxide (P < .0001). This finding is consistent with the purported role of cGMP as the second messenger effecting smooth muscle relaxation in this process. CONCLUSIONS: CPB may be responsible for postoperative dysfunction of the pulmonary endothelial cell and may contribute to postoperative pulmonary hypertension in children. Inhaled nitric oxide is a potent pulmonary vasodilator after CPB with minimal systemic circulatory effects. It may have important diagnostic and therapeutic applications in patients with congenital heart disease. PMID- 8222108 TI - Influence of serum cholesterol and other coronary risk factors on vasomotion of angiographically normal coronary arteries. AB - BACKGROUND: It has been shown that there is impairment of the vasodilatory response to acetylcholine in patients with hypercholesterolemia and angiographically normal coronary arteries. Moreover, in patients with angiographically smooth coronary arteries, the number of coronary risk factors is associated with a loss of endothelium-dependent vasodilation. The purpose of the present analysis was to evaluate in patients with and without coronary artery disease coronary vasomotor response to dynamic exercise in angiographically normal and stenosed coronary arteries and to relate the response to serum cholesterol levels as well as to other coronary risk factors. METHODS AND RESULTS: Luminal area change during exercise (delta-ex, percent change compared with rest = 100%) was determined by biplane quantitative coronary arteriography in three groups: Group 1 consisted of 14 patients with normal total serum cholesterol of < 200 mg/100 mL; mean, 173 mg/100 mL (mean age, 51 years). Group 2 comprised 23 patients with a slightly elevated cholesterol of 200 to 250 mg/100 mL; mean, 223 mg/100 mL (mean age, 53 years). Group 3 had 24 patients with markedly elevated cholesterol of > 250 mg/100 mL; mean, 288 mg/100 mL (mean age, 54 years). Serum cholesterol levels and categorical risk factors such as positive family history, history of hypertension, smoking, obesity, and diabetes were related to exercise-induced vasomotor response. The three groups did not differ with regard to clinical characteristics, exercise work load, and hemodynamic data measured during exercise. However, delta-ex in normal vessels was significantly different between all three groups (ANOVA, P < .01): +31% (group 1), +18% (group 2), and +4% (group 3). Delta-ex in stenotic vessels did not differ between the groups: -5% (group 1), -13% (group 2), and -12% (group 3). Delta-ex of the nonstenosed vessel correlated significantly and inversely with total cholesterol, with low-density lipoprotein cholesterol, with the ratio of total to high-density lipoprotein cholesterol, and with the number of coronary risk factors present in a patient. High total cholesterol and a history of hypertension were independent risk factors for impaired coronary vasomotion. CONCLUSIONS: In patients with and without coronary artery disease, hypercholesterolemia and a history of hypertension independently impair exercise-induced coronary vasodilation in angiographically normal coronary arteries. In the stenotic vessel, vasomotion during exercise does not appear to be influenced by the actual serum cholesterol. The precise mechanism by which the impaired vasomotion of the angiographically normal coronary arteries is mediated is unknown, but a direct negative effect of hypercholesterolemia on endothelial function or early undetected atherosclerosis appears to be the most likely explanation. PMID- 8222109 TI - Cigarette smoking is associated with dose-related and potentially reversible impairment of endothelium-dependent dilation in healthy young adults. AB - BACKGROUND: Cigarette smoking is the most important modifiable risk factor for atherosclerosis. Endothelial dysfunction is an early event in atherogenesis, and we hypothesized that smoking might be associated with endothelial damage in the systemic arteries of otherwise healthy young adults. METHODS AND RESULTS: We studied noninvasively the brachial arteries of 200 subjects aged 15 to 57 years, all normotensive, nondiabetic with cholesterol level < or = 240 mg/dL and no family history of premature vascular disease: 80 control subjects aged 16 to 56 years (mean, 35), 80 current smokers aged 15 to 55 years (mean, 33), and 40 former smokers aged 25 to 57 years (mean, 38). Total lifetime amount smoked varied from 1 to 75 pack years in the smokers. Using high-resolution ultrasound, vessel diameter was measured at rest, during reactive hyperemia (with flow increase causing endothelium-dependent dilation), and after sublingual glyceryl trinitrate (GTN, an endothelium-independent vasodilator). Flow-mediated dilation (FMD) was observed in all the control subjects (10 +/- 3.3%; range, 4% to 22%) but was impaired or absent in the smokers (4 +/- 3.9%; range, 0% to 17%; P < .0001). FMD in the smokers was inversely related to lifetime dose smoked (6.6 +/- 4.0% in very light smokers, 4.0 +/- 3.1% in light smokers, 3.2 +/- 3.2% in moderate smokers, and 2.6 +/- 1.2% in heavy smokers; P < .01). FMD for the former smokers was 5.1 +/- 4.1% (range, 0% to 15%). In a multivariate model adjusting for age, sex, cholesterol, smoking history, and vessel size, former smoking was associated with a higher FMD than current smoking (P = .07); when only male former and current smokers were considered, the higher FMD was significant (P = .0001) but not for female smokers (P = .24). GTN caused dilation in all subjects (control subjects, 20 +/- 5.2%; smokers, 17 +/- 5.8%; former smokers, 17.4 +/- 5.4%). Vessel diameter, baseline flow, and degree of reactive hyperemia (Doppler estimated) were similar in all groups. CONCLUSIONS: Cigarette smoking is associated with dose-related and potentially reversible impairment of endothelium dependent arterial dilation in asymptomatic young adults, consistent with endothelial dysfunction. PMID- 8222110 TI - Cigarette smoking and the development and progression of aortic atherosclerosis. A 9-year population-based follow-up study in women. AB - BACKGROUND: Cigarette smoking has been recognized as an important risk factor for cardiovascular disease in men and women. Whether the increased risk results from an atherogenic effect of smoking is still debated. We examined the relation between cigarette smoking and atherosclerotic changes in the abdominal aorta. METHODS AND RESULTS: The association between cigarette smoking and atherosclerotic changes in the abdominal aorta was examined in a population-based cohort of 758 women, initially aged 45 to 64 years. All women were examined radiographically for calcified deposits in the abdominal aorta, which have been shown to represent intimal atherosclerosis. After 9 years of follow-up, atherosclerotic changes, indicating development or progression of plaques, could be demonstrated in 37% of women. A direct association was found between atherosclerotic change and number of cigarettes smoked per day. Compared with women who had never smoked, the relative risks of those who smoked 1 to 9, 10 to 19, and 20 or more cigarettes per day were 1.4 (95% confidence interval, 1.0 to 2.0), 2.0 (1.6 to 2.5), and 2.3 (1.8 to 3.0), respectively, after adjustment for age and other cardiovascular risk factors. Associations of atherosclerotic change with inhaling habit and duration of smoking were borderline significant after number of cigarettes smoked per day was taken into account. Among former smokers, the risk decreased with increasing duration of stopping but a significant excess risk was still observed after 5 to 10 years since quitting (relative risk, 1.6; 95% confidence interval, 1.1 to 2.2). CONCLUSIONS: These follow-up data support the evidence for an effect of cigarette smoking on atherosclerosis. The findings suggest that the rate of atherosclerotic change may be reduced by cessation of smoking, but a residual effect appears to be present for at least 10 years. PMID- 8222111 TI - Associations of postmenopausal estrogen use with cardiovascular disease and its risk factors in older women. The CHS Collaborative Research Group. AB - BACKGROUND: Postmenopausal estrogen replacement therapy has been associated with favorable levels of cardiovascular disease risk factors, but these associations and the relations between estrogen use and subclinical disease have not been examined in large samples of older women. METHODS AND RESULTS: Present and past estrogen use was ascertained in 2955 women > or = 65 years old in the Cardiovascular Health Study, a study of risk factors for coronary heart disease and stroke in the elderly. Present estrogen use was reported by 12% of these women and past use by an additional 26.5%. Estrogen use (past or present) was strongly associated with lower low-density lipoprotein cholesterol, fibrinogen, glucose, insulin, obesity, and age and higher high-density lipoprotein cholesterol and socioeconomic status (all P < .0001). Estrogen users also had lower levels of subclinical disease as measured by carotid intimal-medial thickness, carotid stenosis grade, ECG left ventricular mass, and Doppler mitral peak flow velocities (each P < .02). Relations were similar in younger and older women (65 to 74 versus > or = 75 years) and smokers and nonsmokers and were unchanged after women with poor medication compliance were excluded. After adjustment for other factors, estrogen use was associated with decreased carotid wall thickness, although this association was of borderline significance after further adjustment for lipids. CONCLUSIONS: Postmenopausal estrogen use in this sample of older women was associated with favorable cardiovascular disease risk factor profiles and with lower measures of subclinical disease. These findings suggest that postmenopausal estrogen use may be associated with lower risk of cardiovascular disease in women well into the eighth decade of life. PMID- 8222112 TI - Socioeconomic status and coronary heart disease risk factor trends. The Minnesota Heart Survey. AB - BACKGROUND: Socioeconomic status (SES) indicators including education, income, and occupation are associated with coronary heart disease (CHD) risk factors, morbidity, and mortality. In most industrialized nations, individuals with less education, lower income, and blue collar occupations have the highest CHD rates. It is suggested by some that these differences by SES are increasing even as age adjusted CHD mortality declines. METHODS AND RESULTS: The Minnesota Heart Survey includes measurement of CHD risk factors and behaviors in population-based samples of Minneapolis-St. Paul adults aged 25 to 74 years in 1980 to 1982 (N = 3243) and 1985 to 1987 (N = 4538). Education was significantly and inversely related to blood pressure, cigarette smoking, body mass index, and a summary risk score for both men and women. Serum cholesterol was inversely related to education in women but not in men. Education was positively associated with leisure physical activity and health knowledge. Associations with household income were less consistent in magnitude and direction. Risk characteristics improved significantly between the 1980 to 1982 and 1985 to 1987 surveys. These changes were similar across education and household income levels. CONCLUSIONS: Improvement in CHD risk factors over time unrelated to education or income suggests that population-wide factors such as improved health knowledge, availability of healthy food items, hypertension treatment, and restrictions on cigarette smoking are operating beneficially in all SES groups. Although the SES gradient in risk factors is not increasing, it remains substantial and indicates directions for future prevention efforts. PMID- 8222113 TI - Relation of plasma levels and composition of apolipoprotein B-containing lipoproteins to angiographically defined coronary artery disease in young patients with myocardial infarction. AB - BACKGROUND: Hypertriglyceridemia is a common metabolic disturbance in men < 45 years old with myocardial infarction. To further investigate the relation between triglyceride-rich lipoproteins and severity of coronary atherosclerosis in this subset of postinfarction patients, apolipoprotein B-containing lipoproteins of 64 consecutive patients were subfractionated in connection with coronary angiography. METHODS AND RESULTS: Density-gradient ultracentrifugation of plasma and coronary angiography were performed 4 to 6 months after the myocardial infarction. Global coronary atherosclerosis and the number and severity of distinct stenoses were evaluated by semiquantitative analysis of 15 proximal coronary segments. The majority of the patients (60%) were hypertriglyceridemic and had higher coronary scores than normotriglyceridemic patients. Of the major plasma lipoproteins, triglycerides and cholesterol in the low-density lipoprotein (LDL) fraction were associated with global coronary atherosclerosis, whereas LDL triglycerides and high-density lipoprotein (HDL) cholesterol correlated directly and inversely, respectively, with the coronary stenosis score. Plasma apolipoprotein B correlated with both coronary scores. The plasma concentrations of lipid and protein in the very-low-density lipoprotein (VLDL) subfractions (VLDL1 through VLDL3) and intermediate-density lipoprotein (IDL) did not correlate with either of the coronary scores, whereas the concentration of triglycerides in dense LDL (density > 1.040 kg/L) was strongly associated with both coronary scores. Compositional analysis of the smallest VLDL particles (VLDL3) and IDL revealed a correlation between the number of cholesteryl ester molecules in small VLDL and global coronary atherosclerosis in hypertriglyceridemic patients. CONCLUSIONS: Global coronary atherosclerosis and distinct stenoses in young postinfarction patients are associated with the number of apolipoprotein B-containing particles in plasma and the concentration of LDL triglyceride. Specifically, dense triglyceride-rich LDL particles and, in hypertriglyceridemic patients, small cholesteryl ester-rich VLDL particles relate to coronary artery disease severity. PMID- 8222114 TI - Alcohol consumption and insulin concentrations. Role of insulin in associations of alcohol intake with high-density lipoprotein cholesterol and triglycerides. AB - BACKGROUND: The relation between alcohol intake and insulin levels may explain, in part, the reported associations of alcohol with cardiovascular disease risk factors, including high-density lipoprotein (HDL) cholesterol, triglycerides, blood pressure, and glucose levels, each of which has been recognized as a component of the insulin resistance syndrome. METHODS AND RESULTS: Subjects included nondiabetic participants of the Kaiser Permanente Women Twins Study (1989 through 1990). Usual alcohol intake was assessed as part of a food frequency questionnaire. For women from twin pairs in which both twins drank (n = 338), an increment of 12 g of alcohol per day (about one drink) was associated with an 8% lower 2-hour post-glucose-load insulin (P < .01) in a multiple regression analysis for twin data, adjusted for age, body mass index, waist-to hip ratio, total caloric intake, and family history of diabetes. With genetic influences removed by matched analysis of the subset of 98 monozygotic twin pairs, an intrapair difference of 12 g of alcohol per day was associated with a 12.4% intrapair decrement in postload insulin (P < .01). Inverse associations were also seen for fasting insulin. Alcohol consumption was inversely associated with postload glucose but not with fasting glucose in unmatched (P = .05) and matched (P = .005) analyses. A significant positive association of alcohol intake with high-density lipoprotein cholesterol and an inverse relation of alcohol intake with triglycerides were each independent of insulin levels (P < or = .02 in the matched models). Neither systolic nor diastolic blood pressures were related to alcohol consumption in this sample, perhaps because of the rather low level of alcohol intake in the study population (median, 4 g/d). CONCLUSIONS: Within the range of light to moderate drinking habits, alcohol consumption was inversely related to fasting and postload insulin levels. This relation did not explain associations of alcohol intake with lipid levels and may instead reflect an additional mechanism by which moderate alcohol consumption impacts cardiovascular disease risk. PMID- 8222115 TI - Does endomyocardial biopsy aid in the diagnosis of active rheumatic carditis? AB - BACKGROUND: Carditis is the only component of rheumatic fever that leads to permanent disability. The diagnosis of carditis is presently made by using composite clinical criteria based on the revised Jones' criteria. Since myocardial involvement is an important component of rheumatic carditis, right ventricular endomyocardial biopsies were performed in 54 patients with clinical acute rheumatic fever and quiescent rheumatic heart disease to evaluate the role of biopsy for the diagnosis of rheumatic carditis. METHODS AND RESULTS: In 11 of the 54 patients, clinical consensus was certain about rheumatic fever and carditis based on the revised Jones' criteria (group 1). Histomorphological abnormalities in these patients were scarce. The diagnostic features of rheumatic myocarditis including Aschoff nodules or histiocytic aggregates were encountered in 3 patients (27%). Lymphocytic infiltration was sparse. A majority of patients demonstrated myocyte degeneration, interstitial degeneration, or occasional interstitial mononuclear cell infiltration, but since these histopathological lesions may occur in other conditions also, they were considered nondiagnostic. In 33 of the 54 patients with preexisting rheumatic heart disease, the diagnosis of carditis was suspected based on varied clinical presentations. Since previous cardiac findings were not available in these patients, the clinical diagnosis of carditis could not be made without equivocation (group 2). Twenty-three patients presented with unexplained acute onset of congestive heart failure and evidence of recent streptococcal infection (group 2A). While 13 of them had one or more other major manifestations, 10 patients had only minor manifestations. Mimetic carditis was suspected in the remaining 10 of 33 patients based on carditis having occurred in previous episodes of rheumatic fever (group 2B). The endomyocardial biopsy provided confirmatory evidence of rheumatic myocarditis in 9 patients of group 2A but in none of the 10 patients with suspected mimetic carditis. Nondiagnostic myocyte or interstitial alterations were frequently observed in group 2. Ten of the 54 patients had no clinical evidence of active carditis (group 3). No histological alterations diagnostic of rheumatic carditis were noted in these patients. Twenty-two follow-up biopsies were performed in the first 10 consecutive patients. Diagnostic histiocytic aggregates or Aschoff nodules were observed in initial biopsies in 4 of 10 patients, and nonspecific myocyte or interstitial alterations were observed in 9. All patients with diagnostic changes in initial biopsy demonstrated fibrohistiocytic nodules in 6- or 12-week biopsy samples. Nondiagnostic alterations, similar to those seen in acute cases, were present in 5 of 8 patients at 6 weeks, 5 of 8 patients at 12 weeks, and 3 of the 6 patients at 24 weeks despite the presumed adequate immunosuppressive therapy. No complications related to biopsy were encountered. CONCLUSIONS: The present study highlights the low frequency of diagnostic features in the biopsy specimens of patients with definite clinical rheumatic carditis. Although such alterations are not observed in patients with chronic rheumatic heart disease, endomyocardial biopsy does not appear to provide additional diagnostic information where clinical consensus is certain about diagnosis of rheumatic carditis. Our study, however, substantiates the concept of carditis underlying unexplained congestive heart failure of acute onset in patients with preexisting rheumatic heart disease and elevated antistreptolysin-O titers. PMID- 8222116 TI - Regional left ventricular mechanics in hypertrophic cardiomyopathy. AB - BACKGROUND: Nonuniformity is a determinant of diastolic function. In patients with hypertrophic cardiomyopathy, hypertrophy, abnormal calcium handling, and regional ischemia can also play a role. This study was designed to assess regional mechanics, asynchrony, and asynergy in patients with hypertrophic cardiomyopathy. METHODS AND RESULTS: Nine control subjects and 22 patients with hypertrophic cardiomyopathy were studied by biplane left ventriculography and high-fidelity pressure tracings for the assessment of diastolic function by computing the time constant of isovolumic relaxation, peak filling rate, and the constant of passive chamber stiffness. Regional mechanics were evaluated by dividing the left ventricle into six sectors in the right and left anterior oblique projections. Systolic and diastolic asynchrony were assessed from the coefficient of variation of the regional time intervals from end diastole to end systole and to peak filling rate, respectively. Asynergy was evaluated from the coefficient of variation of the regional area reduction. Regional passive elastic properties were estimated by computing the regional constant of chamber stiffness. In patients with hypertrophic cardiomyopathy, isovolumic relaxation was prolonged (time constant of isovolumic relaxation 101 +/- 41 versus 51 +/- 16 milliseconds in control subjects; P < .001) and the constant of chamber stiffness was increased (0.056 +/- 0.038 versus 0.025 +/- 0.010 mL-1; P < .001). Both systolic and diastolic asynchrony as well as asynergy were found. Regional mechanics showed hyperkinesia in the free wall, whereas the septum exhibited normal wall motion and increased constant of chamber stiffness. CONCLUSIONS: Diastolic function is impaired in hypertrophic cardiomyopathy, and such an impairment is the consequence of nonuniformity and hypertrophy. The regions where the myopathic process is more pronounced show normal wall motion but increased stiffness. The inhomogeneity of regional wall motion with regional hyperkinesia and normokinesia of neighboring regions results in left ventricular asynergy. PMID- 8222117 TI - Impact of regional ventricular function, geometry, and dobutamine stress on quantitative 99mTc-sestamibi defect size. AB - BACKGROUND: Serial myocardial perfusion imaging with 99mTc-sestamibi (MIBI) was used to evaluate infarct risk area and salvage after thrombolysis. The purpose of this investigation was to determine whether changes in MIBI defect size observed after reperfusion may result in part from distortion of regional and global left ventricular geometry. METHODS AND RESULTS: Twenty-five open-chest dogs were subjected to either 15 minutes (groups 1A and 1B) or 3 hours (group 2) of left anterior descending coronary artery occlusion followed by 3 hours of reperfusion. MIBI was injected before occlusion (group 1A) or during occlusion (groups 1B and 2), and serial ECG-gated planar imaging was performed. Dobutamine was infused after 3 hours of reperfusion (groups 1B and 2) to transiently alter left ventricular size and function. Perfusion defect magnitude (DM) and extent (DE) were serially quantified with circumferential profile analysis of end-systolic (ES), end-diastolic (ED), and summed images. Flow was assessed with radiolabeled microspheres and correlated with myocardial MIBI activity. Myocardial thickening was assessed in the risk area with sonomicrometers. In group 1A dogs, ischemic dyskinesis produced large artifactual quantitative MIBI defects on ES images (DM, 9.3 +/- 1.3; DE, 27.8 +/- 6.0) that were significantly smaller on ED images (DM, 4.5 +/- 0.9, P < .05; DE, 4.4 +/- 2.3, P < .05). In addition, DM and DE correlated inversely with myocardial thickening on ES images (DM, r = -.84; DE, r = -.78) and summed images (DM, r = -.72; DE, r = -.61) but not ED images (DM, r = -.12; DE, r = -.15). An index of defect reduction derived from summed images correlated well with thickening fraction in stunned dogs (group 1B, r = .89) but poorly in infarcted dogs (group 2, r = .41) subjected to dobutamine stress. CONCLUSIONS: 99mTc-MIBI defect size may be affected by alteration of left ventricular geometry. Changes in regional function may confound analysis of risk area and myocardial salvage with serial 99mTc-MIBI imaging and may also affect defect size during pharmacological stress with dobutamine. Dobutamine 99mTc-MIBI imaging may be useful for distinguishing viable and nonviable myocardium. PMID- 8222118 TI - Helical and retrograde secondary flow patterns in the aortic arch studied by three-directional magnetic resonance velocity mapping. AB - BACKGROUND: Helical and retrograde secondary flows have been recorded in the aorta, but their origins and movements in relation to the arch have not been clarified. We set out to do this using magnetic resonance velocity mapping. METHODS AND RESULTS: Three-directional phase contrast cine magnetic resonance velocity mapping was used to map multidirectional flow velocities in the aortas of 10 healthy volunteers. Computer processing was used to visualize flow vector patterns in selected planes. Right-handed helical flows predominated in the upper aortic arch in late systole, being clearly recognizable in 9 of the 10 subjects. Nonaxial components of velocity in this region reached 0.29 m/s (+/- 0.05 m/s) as axial velocities declined from a peak of 1.0 m/s (+/- 0.1 m/s). Helical flow patterns in the upper descending aorta varied between subjects, apparently depending on arch curvature. End-systolic retrograde flow originated from regions of blood with low momentum, usually along inner wall curvatures. Flow studies in a curved tubular phantom showed right-handed helical flow in the upper "arch" when the inflow section was positioned to simulate ascending aortic curvature, and retrograde flow occurred along the inner wall at end systole during pulsatile flow. CONCLUSIONS: Helical and retrograde streams are consistent features of intra-aortic flow in healthy subjects that result, at least in part, from the curvature of the arch and the pulsatility of flow in it. They may have significance in relation to circulatory dynamics and the pathogenesis of atheroma in the arch. PMID- 8222119 TI - Influence of right heart size on outcome in pulmonary atresia with intact ventricular septum. AB - BACKGROUND: Neonates with pulmonary atresia and intact ventricular septum (PA IVS) are frequently born with hypoplastic right heart structures that must grow after right ventricular decompression (RVD) procedures for a complete two ventricle physiology to be achieved. Previous authors have asserted that neonatal right heart size or morphology will predict right heart growth potential. Since 1983, our bias has favored early RVD regardless of initial right heart size. In 1986, we recognized a subset of patients with coronary artery abnormalities associated with poor outcome after RVD and have defined these patients as having a right ventricular-dependent coronary circulation (RVDCC). METHODS AND RESULTS: To assess the influence of right heart size on outcome independent of the presence of RVDCC, we measured echocardiographic right ventricular (RV) dimensions in 37 neonates with adequate studies presenting between 1983 and 1990. Coronary artery anatomy was adequately assessed by angiography in 36. RV volume and tricuspid valve (TV) diameter were significantly smaller in patients with RVDCC than in those without. However, there was no statistically significant association between RV volume or TV diameter and survival among patients with or without RVDCC: Among 29 patients without RVDCC, 23 of 24 (95.8%) who achieved RVD are alive compared with 1 of 5 (20%) who did not achieve RVD (P = .001). Twenty one of the 23 survivors have a complete two-ventricle physiology with low right atrial pressure. Among 7 patients with RVDCC, 2 patients who underwent RVD died early of left ventricular failure, whereas 4 of 5 who did not undergo RVD have survived single ventricular palliation. CONCLUSIONS: Small right heart size is associated with RVDCC but is not associated with survival in PA-IVS. Patients without RVDCC have improved survival after RVD regardless of neonatal right heart size. PMID- 8222120 TI - Pulmonary regurgitation in the late postoperative follow-up of tetralogy of Fallot. Volumetric quantitation by nuclear magnetic resonance velocity mapping. AB - BACKGROUND: Pulmonary regurgitation frequently occurs after surgical correction of tetralogy of Fallot. To date, reliable quantitation of pulmonary regurgitation has not been possible, and therefore the clinical significance of pulmonary regurgitation is controversial. Nuclear magnetic resonance (NMR) velocity mapping allows accurate measurement of volumetric flow. The feasibility and accuracy of NMR velocity mapping to quantify pulmonary regurgitation volumes are studied in patients after Fallot repair. METHODS AND RESULTS: In 18 patients (mean age, 16.5 +/- 6.5 years), late (12.6 +/- 5.2 years) after Fallot surgery, forward and regurgitant volume flow was measured in the main pulmonary artery with NMR velocity mapping. To validate the measurements of pulmonary forward flow, right ventricular stroke volume was used as an internal reference standard. Pulmonary regurgitation volumes were compared with the differences between the corresponding right and left ventricular stroke volumes. Ventricular volumes were measured with a multisection gradient echo NMR method. In addition, the relation between pulmonary regurgitation and right ventricular volumes was studied. Measurements of pulmonary regurgitation volume with NMR velocity mapping closely corresponded with the tomographically determined volumes (r = .93). Forward pulmonary volume flow was nearly identical to right ventricular stroke volume (r = .98). Pulmonary regurgitation volume was significantly correlated with end diastolic volume (r = .82, P < .0005), end-systolic volume (r = .63, P < .01), and stroke volume (r = .89, P < .0005) of the right ventricular but not with right ventricular ejection fraction (r = .41, P = NS). CONCLUSIONS: NMR velocity mapping is an accurate method for the noninvasive, volumetric quantification of pulmonary regurgitation after surgical correction of tetralogy of Fallot. PMID- 8222121 TI - Effect of inotropic stimulation on the negative force-frequency relationship in the failing human heart. AB - BACKGROUND: In severe human heart failure, an increase in frequency of stimulation is accompanied by a reduced force of contraction in vivo and in vitro. The present study was aimed to investigate whether inotropic stimulation influences the inverse force-frequency relationship in failing human myocardium. METHODS AND RESULTS: The effects of the cAMP-independent positive inotropic agents ouabain (0.01 mumol/L) and BDF 9148 (0.1 mumol/L) as well as the beta adrenoceptor agonist isoprenaline (0.01 mumol/L and 0.1 mumol/L) on the force frequency relationship in electrically driven papillary muscle strips from nonfailing (brain death, n = 5) and terminally failing (NYHA class IV, heart transplants, dilated cardiomyopathy, n = 22) human myocardium were studied. For comparison, we examined the effect of elevation of the extracellular Ca2+ concentration (3.2 mmol/L and 6.2 mmol/L). In nonfailing myocardium, force of contraction, peak rate of tension rise, and peak rate of tension decay increased, whereas time to peak tension and time to half relaxation decreased following an increase of stimulation frequency. In NYHA class IV, force of contraction gradually declined followed by changes of other parameters of isometric contraction. Moderate stimulation of contractility by isoprenaline (0.01 mumol/L) partly reversed the negative force-frequency relationship in NYHA class IV and preserved the positive force-frequency relationship in nonfailing myocardium. The addition of ouabain and BDF 9148 together restored completely the force-frequency relationship in NYHA class IV. In contrast, high concentrations of isoprenaline (0.1 mumol/L) and an elevation of the extracellular Ca2+ concentration enhanced the decline in force of contraction in the presence of higher stimulation frequencies. CONCLUSIONS: It is concluded that functionally important changes occur in the intracellular Ca2+ handling, leading to the negative force-frequency relationship in terminally failing human myocardium. Interestingly, the negative force-frequency relationship can be restored by agents producing positive inotropic effects by elevation of the intracellular Na+ concentration. These findings suggest that hitherto unknown changes in the intracellular ionic homeostasis occur in the failing human heart. Even though increasing [Ca2+]i in failing heart cells may be detrimental, increasing [Na+bdi may be beneficial through a mechanism independent of an increase in [Ca2+]i. PMID- 8222122 TI - Effects of the angiotensin converting enzyme inhibitor enalapril on the long-term progression of left ventricular dilatation in patients with asymptomatic systolic dysfunction. SOLVD (Studies of Left Ventricular Dysfunction) Investigators. AB - BACKGROUND: Patients with heart failure and reduced left ventricular (LV) ejection fraction (EF) manifest progressive LV dilatation, which is prevented by angiotensin converting enzyme (ACE) inhibitors. In patients with asymptomatic LV systolic dysfunction, in whom there is less activation of the renin-angiotensin system, ventricular remodeling might be less rapid and the benefit of ACE inhibitors less discernible. METHODS AND RESULTS: One hundred eight patients enrolled in the Studies of Left Ventricular Dysfunction (SOLVD) Prevention Trial, with left ventricular ejection fraction < or = 0.35 but without clinical heart failure, underwent radionuclide ventriculograms, and 49 underwent left heart catheterizations. Measurements were made before and after double-blinded randomization to enalapril (2.5 to 20 mg/d) or placebo. Repeated-measures analysis of all time points showed significant differences for change in end diastolic volume (EDV) between enalapril and placebo groups. Significant difference between the enalapril and placebo groups (P < .05) was present for change in EDV at 1 year within the catheterization study and at a mean of 25 months within the radionuclide study. Radionuclide EDV increased in placebo patients (119 +/- 28 to 124 +/- 33 mL/m2, mean +/- SD) and decreased in enalapril patients (120 +/- 25 to 113 +/- 25 mL/m2). Differences between the two groups were significantly less than previously described in patients with symptomatic heart failure (P < .02), with less increase in LV volumes in the placebo group and less decrease in volumes in the enalapril group. CONCLUSIONS: Chronic ACE inhibitor treatment slows or reverses LV dilatation in patients with asymptomatic LV systolic dysfunction. Compared with symptomatic patients, asymptomatic patients manifest a slower rate of spontaneous LV dilatation and less reduction in LV volumes by enalapril. PMID- 8222123 TI - Circadian variation of sudden cardiac death reflects age-related variability in ventricular fibrillation. AB - BACKGROUND: Previous studies report a morning peak in the occurrence of out-of hospital sudden cardiac death but lack detailed information on underlying arrhythmias. We used the documentation system of the semiautomated defibrillators used by emergency medical technicians to investigate the circadian pattern of defined arrhythmias and the influence of demographic patient characteristics on this pattern. METHODS AND RESULTS: From December 1988 to December 1990, 703 consecutive patients (63% men; age, 67 +/- 17 years) with sudden cardiac death were registered in the Klinikum Steglitz area of the Berlin emergency care system. Determination of time of day of the event was based on the arrival time of the rescue squad. A marked circadian variation (P < .0001) in the occurrence of sudden cardiac death was observed with a primary morning peak (6 AM to noon) and a secondary afternoon peak (3 to 7 PM). The subgroup of 294 patients with ventricular fibrillation as initially documented arrhythmia showed a similar circadian variation (P < .0001). In significant contrast (P < .01), patients with asystole (n = 260) or pulseless bradyarrhythmias (n = 149) were more evenly distributed during the daytime with a primary night trough. Multivariate logistic regression analysis revealed that the circadian pattern of ventricular fibrillation was similar in both gender groups but tended to differ with regard to age: patients older than 65 years demonstrated a monophasic distribution, whereas patients aged 65 years or less had a biphasic distribution with peaks in the morning and in the afternoon. CONCLUSIONS: The circadian pattern of sudden cardiac death reflects primarily a circadian variation in onset of ventricular fibrillation. The different circadian patterns of ventricular fibrillation, pulseless bradyarrhythmias, and asystole suggest different pathophysiological mechanisms of causation of death. The age dependence of the pattern of ventricular fibrillation may indicate different underlying external or endogenous triggers. PMID- 8222124 TI - Localization of the site of origin of postinfarction ventricular tachycardia by endocardial pace mapping. Body surface mapping compared with the 12-lead electrocardiogram. AB - BACKGROUND: The purpose of this study was to assess the value of body surface mapping and the standard 12-lead ECG in localizing the site of origin of postinfarction ventricular tachycardia (VT) during endocardial pace mapping of the left ventricle. METHODS AND RESULTS: Simultaneous recordings of 62-lead body surface QRS integral maps and scalar 12-lead ECG tracings were obtained in 16 patients with prior myocardial infarction during a total of 26 distinct VT configurations and during subsequent left ventricular catheter pace mapping at 9 to 24 different endocardial sites. Anatomic pacing site locations were computed by means of a biplane cineradiographic method and plotted on a polar projection of the left ventricle. The QRS integral map and the QRS complexes of the 12 standard leads of each VT morphology obtained in a particular patient were compared independently with the different paced QRS integral maps and paced QRS complexes of the 12-lead ECG generated in that same patient. The stimulus site locations of the best matching paced QRS integral map and paced QRS complexes of the 12-lead ECG were indicated on the polar projection and subsequently compared with the endocardial location of the corresponding site of VT origin identified during intraoperative (surgical ablation) or catheter activation sequence mapping (catheter ablation). The localization resolution of pace mapping was established separately for each electrocardiographic technique by computing the size of endocardial areas with similar morphological features of the QRS complex. Pace mapping advocated with body surface mapping or the 12-lead ECG enabled adequate reproduction of the VT QRS morphology in 24 of 26 VTs (92%) and 25 of 26 VTs (96%), respectively. Activation sequence mapping identified the site of origin in 12 of 26 previously observed VT configurations (46%). Ten and 11 VTs were localized by activation sequence mapping and pace mapping combined with body surface mapping or the 12-lead ECG, respectively. Pace mapping applied with body surface mapping identified the site of origin correctly (distance < or = 2 cm) in 8 of 10 compared VTs (80%); an adjacent site (distance between 2 and 4 cm) or a disparate site (distance > or = 4 cm) was identified in the remaining 2 of 10 VTs (20%). Pace mapping used with the 12-lead ECG localized the site of origin correctly in 2 of 11 VTs (18%); the site of origin was identified correctly next to an additional adjacent site in 5 of 11 VTs (55%); and an adjacent site or a disparate site was found in 1 of 11 VTs (9%) and 2 of 11 VTs (18%), respectively. The difference in localization accuracy of both electrocardiographic techniques was statistically significant (P = .02). The mean size of endocardial areas where a comparable QRS morphology was obtained during pace mapping was 6.0 +/- 4.5 cm2 with the application of body surface mapping and 15.1 +/- 12.0 cm2 with the use of the 12-lead ECG. CONCLUSIONS: These results demonstrate that application of the 62-lead instead of the 12-lead ECG during endocardial pace mapping enhances the localization resolution of this mapping technique and enables more precise identification of the site of arrhythmogenesis in the majority of compared postinfarction VT episodes. PMID- 8222125 TI - High resolution mapping of Koch's triangle using sixty electrodes in humans with atrioventricular junctional (AV nodal) reentrant tachycardia. AB - BACKGROUND: Recent evidence suggests that atrioventricular junctional reentrant tachycardia (AVJRT) uses a reentrant circuit that involves the atrioventricular (AV) node, the atrionodal connections, and perinodal atrial tissue. Electrogram morphology has been used to target the delivery of radiofrequency energy to the site of the "slow pathway," a component of this reentrant circuit. The aim of this study was to localize precisely the sites of atrionodal connections involved in AVJRT and to examine atrial electrogram morphologies and their spatial distribution over Koch's triangle. METHODS AND RESULTS: Electrical activation of Koch's triangle and the proximal coronary sinus was examined in 13 patients using a 60-point plaque electrode and computerized mapping system. Recordings were made during sinus rhythm (n = 12), left atrial pacing (n = 8), ventricular pacing (n = 12), and AVJRT (n = 12). During sinus rhythm electrical activation approached Koch's triangle and the AV node from the direction of the anterior limbus, activating the anterior part of the triangle before the posterior part. A zone of slow conduction during sinus rhythm was found within Koch's triangle in 64% of patients. The pattern of atrial activation in Koch's triangle during anterograde fast pathway conduction was similar to that seen during anterograde slow pathway conduction. Retrograde fast pathway conduction during ventricular pacing and during anterior (typical) AVJRT caused earliest atrial activation at the apex of Koch's triangle near the AV node-His bundle junction. In individual patients the site of earliest atrial activation was similar for both anterior AVJRT and retrograde fast pathway conduction during ventricular pacing. Retrograde slow pathway conduction during ventricular pacing and during posterior (uncommon or atypical) AVJRT caused earliest atrial activation posterior to the AV node near the orifice of the coronary sinus. This posterior or "slow pathway" exit site was 15 +/- 4 mm from the His bundle. In individual patients the site of earliest atrial activation was similar for both posterior AVJRT and retrograde slow pathway conduction during ventricular pacing. In one patient anterograde and retrograde conduction occurred via separate slow pathways during AVJRT: Complex atrial electrograms with two or more components were observed near the coronary sinus orifice and in the posterior part of Koch's triangle in all cases. These were categorized as either low or high frequency potentials according to the rapidity of the second component of the electrogram. Low frequency potentials were present at the site of earliest atrial excitation during retrograde slow pathway conduction in 5 of 5 cases (100%) and high frequency potentials in 4 of 5 cases (80%). However, both slow and high frequency potentials could be found at sites up to 16 mm from the site of earliest atrial excitation. CONCLUSIONS: At least two distinct groups of atrionodal connections exist. The site of earliest atrial activation during anterior AVJRT is similar to that of fast pathway conduction during ventricular pacing. This site is close to the His bundle-AV node junction. The site of earliest atrial activation during posterior AVJRT is similar to that of slow pathway conduction during ventricular pacing. This site is near the coronary sinus orifice, approximately 15 mm from the His bundle. The anterograde slow pathway appears to be different from the retrograde slow pathway in some patients. Double atrial electrograms are an imprecise guide to the site of earliest atrial excitation during retrograde slow pathway conduction. PMID- 8222126 TI - Orthodromic capture of the atrial electrogram during transient entrainment of atrioventricular nodal reentrant tachycardia. AB - BACKGROUND: The reentry circuit of atrioventricular nodal reentrant tachycardia (AVNRT) has not been fully demonstrated. We hypothesized that if an upper common pathway was present, the atrial electrogram could not be captured orthodromically during transient entrainment of AVNRT by rapid atrial pacing. Based on this hypothesis, the presence of an upper common pathway was investigated. METHODS AND RESULTS: The atrial electrogram at the recording site of the His bundle potential was identified during induced AVNRT in 9 patients. To entrain AVNRT transiently, rapid pacing from the high right atrium and coronary sinus was applied at a cycle length 10 milliseconds shorter than that of AVNRT and repeated after a decrement of the paced cycle length in steps of 5 milliseconds until AVNRT was interrupted. In 5 of 7 patients, orthodromic capture of the atrial electrogram at the recording site of the His bundle potential was observed during transient entrainment of AVNRT by coronary sinus pacing, ie, the first postpacing interval of the atrial electrogram at the recording site of the His bundle potential was the same as the paced cycle length. In these 5 patients, the mean minimum paced cycle length capable of orthodromic atrial capture was 349 milliseconds, and the mean difference from the cycle length of AVNRT was only 16 milliseconds. During transient entrainment of AVNRT by high right atrial pacing, the atrial electrogram could not be captured orthodromically. CONCLUSIONS: Observation of orthodromic capture of the atrial electrogram at the recording site of the His bundle potential by coronary sinus pacing ruled out the presence of an upper common pathway in AVNRT, and the concept that perinodal atrial tissue is involved in the reentry circuit of AVNRT was supported. PMID- 8222127 TI - Antineutrophil and myocardial protecting actions of a novel nitric oxide donor after acute myocardial ischemia and reperfusion of dogs. AB - BACKGROUND: It has recently been demonstrated that myocardial ischemia and reperfusion results in a marked decrease in the release of nitric oxide (NO) by the coronary endothelium. NO may possess cardioprotective properties, possibly related to inhibition of neutrophil-related activities. We tested the hypothesis that a cysteine-containing nitric oxide donor compound, SPM-5185, would reduce infarct size and inhibit neutrophil-related activities (adherence to coronary vascular endothelium, accumulation). METHODS AND RESULTS: The effects of intracoronary infusion of SPM-5185 were investigated in a 5.5-hour model of myocardial ischemia (1 hour) and reperfusion (4.5 hours) (MI-R) in anesthetized, open-chest dogs. SPM-5185 (500 nmol/L) or saline vehicle was infused for 4.5 hours into the left anterior descending coronary artery (LAD) at the time of reperfusion after 1 hour of LAD occlusion. MI-R in dogs receiving saline vehicle resulted in severe myocardial injury characterized by dyskinesis, a profound elevation of plasma creatine kinase, marked myocardial necrosis, and high cardiac myeloperoxidase (MPO) activity in the ischemic and necrotic zones. In contrast, treatment with SPM-5185 resulted in a modest restoration of regional function, a reduction of myocardial necrosis expressed as a percentage of the area at risk (12.5 +/- 3.2% versus 41.7 +/- 5.4%, P < .001), and significant reductions of MPO activity in the ischemic zone (0.8 +/- 0.1 versus 2.5 +/- 0.7 U/100 mg tissue, P < .05) and the necrotic zone (1.6 +/- 0.2 versus 3.3 +/- 0.6 U/100 mg tissue, P < .05). In additional studies, SPM-5185 (500 nmol/L) significantly (P < .001) attenuated the adherence of LTB4-stimulated canine neutrophils to autologous segments of coronary artery and attenuated the neutrophil-induced contraction of isolated coronary arterial rings. CONCLUSIONS: SPM-5185 reduces myocardial necrosis and neutrophil accumulation in an acute model of canine myocardial ischemia and reperfusion. This reduction in myocardial cell injury may be partially related to the inhibitory actions of this novel NO donor on neutrophil adherence to the coronary endothelium. PMID- 8222128 TI - Ischemic preconditioning fails to limit infarct size in reserpinized rabbit myocardium. Implication of norepinephrine release in the preconditioning effect. AB - BACKGROUND: Infarct size reduction by ischemic preconditioning is believed to be mediated by adenosine; however, whether adenosine is the factor responsible for the initiation of this protection remains unknown. It is possible that during preconditioning, adenosine stimulates receptors on presynaptic nerve terminals and retards the release of norepinephrine (NE) during the prolonged ischemia or that NE release during preconditioning augments adenosine production. METHODS AND RESULTS: To test whether the release of NE is involved in the preconditioning phenomenon, rabbits were pretreated with reserpine (5 mg/kg sc, 24 hours before) to deplete presynaptic nerve terminals of NE stores. On the day of the experiment, the rabbits were anesthetized with ketamine-xylazine and instrumented for coronary occlusion. Nonreserpinized animals were used as controls. The control group (n = 7) was subjected to 30 minutes of coronary occlusion and 120 minutes of reperfusion (ischemia-reperfusion) only. The preconditioned group (n = 10) received 5 minutes of preconditioning ischemia and 10 minutes of reperfusion before the prolonged ischemia-reperfusion. Of the reserpinized animals, half (n = 7) received preconditioning before ischemia-reperfusion and the remaining animals (n = 7) did not. At termination of the experiment, an intravenous tyramine challenge (1 mg/kg) was used to confirm NE depletion in reserpinized rabbits. The resulting infarcts were measured with tetrazolium and planimetry. With comparable hemodynamics and areas at risk, infarct size in control animals was 39.8 +/- 2.1% of the risk region. Preconditioned animals showed an expected reduction of infarct size to 14.8 +/- 2.2% of risk region (P < .05 vs control). Of the reserpinized animals, those that received reserpine alone had infarcts that were 38.5 +/- 4.5% of risk region, and those that were preconditioned had infarcts that were 41.4 +/- 3.6% of risk region, which was not significantly different than the control group. CONCLUSIONS: We conclude that preconditioning fails to protect ischemic-reperfused myocardium in reserpinized rabbit myocardium, indicating that the release of NE during either preconditioning or prolonged ischemia is critical to preconditioning mediated protection. PMID- 8222129 TI - Augmentation of endogenous adenosine attenuates myocardial 'stunning' independently of coronary flow or hemodynamic effects. AB - BACKGROUND: Mounting evidence suggests a protective effect of exogenous adenosine in myocardial ischemia and reperfusion. We tested the hypothesis that augmentation of endogenous adenosine levels, achieved by inhibiting adenosine catabolism and washout, is beneficial in postischemic myocardial dysfunction ("stunning"). METHODS AND RESULTS: In phase I of the study, open-chest dogs undergoing a 15-minute coronary artery occlusion and 4 hours of reperfusion received an intracoronary infusion of either saline (controls, n = 23) or 6-(4 nitrobenzyl)-mercapto: purine ribonucleoside (NBMPR, a selective nucleoside transport inhibitor) combined with erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA, a potent adenosine deaminase inhibitor) (EHNA + NBMPR, n = 15) starting 15 minutes before coronary occlusion and ending 15 minutes after the initiation of reflow. Regional myocardial function (assessed as systolic wall thickening) was similar in control and treated groups at baseline and during ischemia. After reperfusion, however, the dogs treated with EHNA + NBMPR exhibited a significant improvement in the recovery of function, which was evident as early as 30 minutes after restoration of flow and was sustained throughout the rest of the reperfusion phase. The enhanced recovery effected by EHNA + NBMPR could not be attributed to nonspecific factors such as differences in collateral flow during occlusion, coronary flow after reperfusion, arterial pressure, heart rate, or other hemodynamic variables. In phase II of the study, the myocardial content of adenine nucleotides and nucleosides was measured by high performance liquid chromatography in myocardial biopsies obtained serially from open-chest dogs undergoing the same protocol used in phase I. There were no significant differences between control (n = 8) and treated (n = 9) dogs with respect to myocardial levels of adenosine triphosphate (ATP) at 30 and 60 minutes after reperfusion, indicating that the beneficial effects of EHNA + NBMPR cannot be ascribed to repletion of ATP stores. Compared with controls, dogs treated with EHNA + NBMPR exhibited a much larger increase in myocardial adenosine (6.07 +/- 1.47 vs 1.03 +/- 0.16 nmol/mg protein, P < .05) and a much smaller increase in inosine (0.52 +/- 0.27 vs 3.04 +/- 0.54 nmol/mg protein, P < .05) at the end of ischemia, such that the inosine-to-adenosine ratio noted in controls was completely reversed (approximately 6:1 vs approximately 1:6, respectively). In the treated group, adenosine levels remained markedly increased compared with controls up to 1 hour after reperfusion. CONCLUSIONS: This study demonstrates that (1) administration of an adenosine deaminase inhibitor plus a nucleoside transport blocker is remarkably effective in augmenting myocardial adenosine levels during regional ischemia and subsequent reperfusion in vivo, (2) this augmentation of adenosine results in a significant and sustained attenuation of myocardial stunning, and (3) the attenuation of stunning is not due to ATP repletion or to nonspecific actions on hemodynamic variables or coronary flow. These findings suggest that endogenous adenosine production during ischemia serves as an important pathophysiological mechanism that protects against myocardial stunning. The results also suggest that augmentation of endogenous adenosine (without exogenous adenosine administration) represents an effective therapeutic approach to the alleviation of reversible postischemic dysfunction. PMID- 8222130 TI - Short-term synchronized retroperfusion before reperfusion reduces infarct size after prolonged ischemia in dogs. AB - BACKGROUND: Previous studies have demonstrated that synchronized coronary venous retroperfusion (SRP) can restore blood flow to the ischemic myocardium, resulting in infarct size reduction and improvement of the left ventricular function. Despite the nutritive blood flow achieved by SRP being relatively limited, SRP has been shown to improve washout of by-products from the ischemic myocardium. The aim of this study was to investigate whether short-term SRP immediately prior to reperfusion would attenuate the deteriorative phenomena following reperfusion. METHODS AND RESULTS: Closed-chest anesthetized dogs underwent 3 hours of left anterior descending coronary artery (LAD) occlusion. The dogs were then randomized into two groups: (1) control group (n = 9), in which the occlusion was immediately followed by 3-hour reperfusion; or (2) SRP group (n = 9), in which SRP was started 3 hours after occlusion and maintained for 30 minutes with sustained occlusion followed by 2.5-hour reperfusion with simultaneous discontinuation of SRP. There were no statistical differences between the groups in global hemodynamics and degree of ischemia measured by radiolabeled microspheres. Myocardial infarct size (triphenyltetrazolium method) expressed as percentage of risk area was significantly smaller in the SRP group (24 +/- 7%, mean +/- SEM) than in the control group (54 +/- 9%). The extent of myocardial hemorrhage expressed as percentage of infarct size was also significantly reduced in the SRP group (3 +/- 2%) compared with the control group (24 +/- 6%). The increase in end-diastolic wall thickness in the ischemic area after reperfusion assessed by two-dimensional echocardiography was significantly less in the SRP group. Blood flow measurements after reperfusion demonstrated the occurrence of no-reflow phenomenon only in the control group. Histological examination revealed extensive myocardial hemorrhages only in the control group, which extended into the nonnecrotic myocardium in four of nine hearts and extensive contraction band necrosis compared with the SRP group. CONCLUSIONS: Short-term SRP prior to reperfusion can reduce infarct size, myocardial hemorrhage, wall swelling, and no reflow phenomenon. The mechanism of this beneficial effect is not clear but might be due to gradual reperfusion and washout of by-products from the ischemic myocardium before fully oxygenated arterial blood reperfusion. PMID- 8222131 TI - Mechanism of selective epicardial activation delay during acute myocardial ischemia in dogs. AB - BACKGROUND: Previous studies have shown that acute myocardial ischemia in the dog results in much greater activation delays in epicardial than endocardial tissue. These results have been interpreted to indicate enhanced sensitivity of epicardial conduction properties to acute ischemia. This study was designed to test the hypothesis that the ischemic epicardial activation delay during supraventricular rhythms is due to slow conduction across the ischemic myocardial wall prior to epicardial activation and not to enhanced epicardial conduction slowing per se. METHODS AND RESULTS: Changes in epicardial and endocardial activation were measured with transmural decapolar needle electrodes during successive 5-minute left anterior descending coronary artery (LAD) occlusions separated by 30-minute reperfusion periods. Occlusions were performed during left atrial pacing, right ventricular pacing with a stimulating electrode located along the longitudinal axis of epicardial fiber orientation in the ischemic zone, or left ventricular pacing in nonischemic tissue located on a line transverse to fiber orientation in the ischemic zone. During both atrial and left ventricular pacing, activation in the ischemic zone began in the endocardium. Epicardial activation resulted from transmural conduction and was markedly delayed compared with endocardial activation during acute myocardial ischemia. During right ventricular stimulation, the ischemic zone epicardium was activated via longitudinal epicardial conduction, and its activation was only slightly delayed by acute ischemia. Epicardial activation mapping was used to assess ischemia induced changes in longitudinal epicardial conduction velocity and to compare them with changes in transmural velocity during atrial or left ventricular pacing. Longitudinal conduction in the ischemic epicardium was slowed by 13 +/- 4% (mean +/- SE) relative to preischemic control values in contrast to transmural conduction, which was slowed 50 +/- 4% by LAD occlusion during atrial pacing and 49 +/- 5% during left ventricular pacing (both P < .001 versus longitudinal epicardial conduction). Transmural activation studies showed that the midmyocardium is the site of most of the ischemic activation delay during transmural propagation. CONCLUSIONS: Epicardial activation is more delayed than endocardial by acute ischemia during supraventricular rhythms in dogs because of slowed conduction across the myocardial wall, not because of enhanced sensitivity of epicardial conduction to depression by acute ischemia. PMID- 8222132 TI - Use-dependent block of the pacemaker current I(f) in rabbit sinoatrial node cells by zatebradine (UL-FS 49). On the mode of action of sinus node inhibitors. AB - BACKGROUND: Zatebradine (UL-FS 49) is a drug with a specific bradycardiac electrophysiological profile. It reduces heart rate by lengthening the duration of diastolic depolarization in the sinoatrial (SA) node. The ionic basis of this action, however, is not clarified. METHODS AND RESULTS: We used the whole-cell patch-clamp technique to study the effects of zatebradine on ionic currents underlying diastolic depolarization of isolated rabbit SA node cells. Low concentrations of zatebradine simultaneously reduced diastolic depolarization rate and the pacemaker current I(f). The drug blocked the pacemaker current, I(f), in a use-dependent manner without causing a shift of its activation curve. At hyperpolarized potentials, unblock of I(f) occurred. Clinically relevant concentrations of the drug have little effect on the L-type calcium current or delayed rectifier potassium current. CONCLUSIONS: This use-dependent block of the If channel can account for most of the pharmacological characteristics of zatebradine and is probably the mechanism of heart rate reduction caused by this agent. Thus, the sinus node inhibitor zatebradine belongs to a new class of "I(f) blockers" with possible advantages over currently available drugs for the treatment of ischemic heart disease. PMID- 8222133 TI - Optical recordings of the effect of electrical stimulation on action potential repolarization and the induction of reentry in two-dimensional perfused rabbit epicardium. AB - BACKGROUND: Prolonged membrane depolarization induced by an electric shock in the heart may produce propagation block leading to repetitive beats. We studied prolonged depolarization and its role in repetitive beats in a thin epicardial layer of endocardially prefrozen arterially perfused rabbit heart. METHODS AND RESULTS: A laser scanner recorded optical action potentials at 63 sites within a 1-cm2 area on the left ventricle of hearts stained with potentiometric fluorescent dye. Pacing (S1) produced propagation across the myofibers; then, a 3 millisecond shock (S2) given in the S1 refractory period produced an electric field that decreased in strength with distance along the fibers. The S2 strengths at the center of the scanned region (C) were 2.1 +/- 0.2 or 5.6 +/- 0.3 V/cm (mean +/- SD, n = 4). Repetitive beats occurred in 50% of hearts when C was 2.1 V/cm and in 100% of hearts when C was 5.6 V/cm. With each occurrence of repetitive beats, prolonged depolarization of the shocked action potential occurred within 1 mm of the S2 electrode when C was 2.1 V/cm and within 3 mm when C was 5.6 V/cm. Transient block immediately after S2 occurred between tissue with prolonged depolarization (S2 strength, 6 to 9 V/cm) and tissue without prolonged depolarization (S2 strength, 1 to 3 V/cm). Propagation in the scanned region after S2 occurred first on the side of the block distal to the S2 electrode, propagated from the most recovered to the least recovered tissue, and then turned toward the S2 electrode. When C was 5.6 V/cm, reentry by retrograde propagation near the S2 electrode produced repetitive beats. The center of the reentrant circuit exhibited further transient block and small depolarizations associated with the circulating activation. CONCLUSIONS: Prolonged depolarization occurs where the S2 strength is more than 6 V/cm, block occurs between regions of prolonged depolarization and no prolonged depolarization, and reentry occurs around the block. Shock-induced prolonged depolarization can be proarrhythmic and may account for electrically induced arrhythmias. PMID- 8222134 TI - Transition from compensated hypertrophy to intrinsic myocardial dysfunction during development of left ventricular pressure-overload hypertrophy in conscious sheep. Systolic dysfunction precedes diastolic dysfunction. AB - BACKGROUND: Patients with aortic stenosis have a period of compensated left ventricular hypertrophy but may eventually develop congestive heart failure. Previous experimental studies showed either normal myocardial contractility in mild short-term pressure overload or myocardial dysfunction with severe pressure overload. Transition from compensated left ventricular hypertrophy to myocardial dysfunction has not been experimentally demonstrated in an adult large animal. Controversial issues in pressure-overload hypertrophy include whether the left ventricular dysfunction is due to insufficient hypertrophy (afterload mismatch) or to intrinsic myocardial dysfunction and whether diastolic dysfunction precedes systolic dysfunction. METHODS AND RESULTS: We induced left ventricular hypertrophy (41% increase in left ventricular to body weight ratio) by gradually tightening a hydraulic constrictor around the ascending aorta in 9 chronically instrumented conscious sheep. Afterload (end-systolic stress) elevation remained constant (approximately 33% greater than baseline) by adjustment of the aortic constrictor over 6 weeks, gradually increasing left ventricular pressure (from 117 +/- 6 to 163 +/- 5 mm Hg) as hypertrophy developed. Four sets (baseline, 2 weeks, 4 weeks, and 6 weeks) of serial hemodynamic studies were performed in each animal with beta-blockade, first with and then without aortic constriction to mechanically match loading conditions. Stepwise methoxamine infusion was performed to obtain load-independent assessment of myocardial contractility. Midwall shortening (P < .05) and shortening rate (P < .05) at mechanically matched loading conditions showed that myocardial dysfunction developed between the fourth and the sixth week. Shortening-preload-afterload (P < .05) and shortening rate-preload-afterload (P < .05) relations, load-independent contractility indices based on the systolic myocardial stiffness concept, also revealed depressed myocardial contractility at the sixth week. Time constant of left ventricular isovolumic relaxation and diastolic myocardial stiffness constant did not change over the 6 weeks. CONCLUSIONS: Transition from normal myocardial contractility to myocardial dysfunction was demonstrated. This transition occurred even when the elevation of afterload remained constant as hypertrophy incompletely adapted to increasing left ventricular pressure. Systolic dysfunction preceded diastolic dysfunction in this model. PMID- 8222135 TI - Potential benefits of late reperfusion of infarcted myocardium. The open artery hypothesis. PMID- 8222136 TI - Images in cardiovascular medicine. Systolic obliteration of the left anterior descending coronary artery. PMID- 8222137 TI - A 49-year-old woman with hypertension who deteriorates after acute myocardial infarction. PMID- 8222138 TI - Cytoprotective effects of nitric oxide. PMID- 8222139 TI - The bailout stent. Is a friend in need always a friend indeed? PMID- 8222140 TI - How to estimate restenosis rate. PMID- 8222141 TI - Human blood pressure determination by sphygmomanometry. PMID- 8222142 TI - Lewis A. Conner Lecture. Contributions of epidemiology to the prevention of stroke. PMID- 8222143 TI - Report of the task force on children and youth. American Heart Association. PMID- 8222144 TI - Management of patients with intramural hematoma of the thoracic aorta. AB - BACKGROUND: Intramural hematoma of the thoracic aorta (IMH) is a diagnosis of exclusion and represents spontaneous, localized hemorrhage into the wall of the thoracic aorta in the absence of bona fide aortic dissection, intimal tear, or penetrating atherosclerotic ulcer. This process may arise from primary vasa vasorum hemorrhage within the aortic media or rupture of an atherosclerotic plaque. The clinical presentation of patients with IMH mimics that of acute aortic dissection; moreover, considerable diagnostic confusion exists despite the use of many different imaging modalities. The optimal mode of management of patients with IMH (medical versus medical plus surgical) remains problematic because of the paucity of information available. METHODS AND RESULTS: Thirteen patients with IMH were managed at two medical centers between 1983 and 1992. Patients with IMH caused by giant penetrating atherosclerotic ulcers were specifically excluded. There were 8 women and 5 men (mean age, 70 years [range, 54 to 82 years]). The admitting clinical diagnosis was acute aortic dissection, and all patients had a history of hypertension. There was no evidence of aortic dissection or intimal disruption as assessed by computed tomographic (CT) scan (n = 11), aortography (n = 10), magnetic resonance imaging (MRI) scan (n = 9), transesophageal echocardiography (TEE) (n = 6), or intravascular ultrasound (n = 1). The diagnosis of IMH was established by exclusion. The descending thoracic aorta was involved in 10 cases and the ascending/arch in 3. Conservative medical management was attempted initially. All 3 patients with IMH involving the ascending aorta ultimately required operative intervention, and 2 individuals died; 2 of 10 patients with descending aortic involvement eventually underwent surgery. Average hospital stay was 11 days; the mean follow-up interval for discharged patients was 29 months. CONCLUSIONS: IMH is a distinct pathological entity, should not be confused with aortic dissection, and probably will be identified more frequently in the future. All patients with IMH should be monitored carefully and treated with aggressive antihypertensive therapy. Frequent serial assessment is necessary using TEE or MRI/CT scans. Based on this small experience, patients with ascending/arch IMH, ongoing pain, or IMH expansion should probably undergo early graft replacement. Patients with IMH involving the descending thoracic aorta who have no evidence of progression and become pain free can probably be treated conservatively but require antihypertensive therapy and serial aortic imaging surveillance indefinitely. PMID- 8222145 TI - Aortic valve replacement in patients 80 years and older. Operative risks and long term results. AB - BACKGROUND: Forty percent of 7.4 million Americans aged 80 years and older have symptomatic heart disease. Controversy exists as to whether the health care resources allotted to this patient subset represent a cost-effective approach to attaining a meaningful quality of life. Although aortic valve surgery carries greater risks in older than in younger patients, published studies reveal that the elderly should not be denied this procedure. METHODS AND RESULTS: To determine the results of aortic valve replacement (AVR) in an elderly population, we retrospectively analyzed 171 consecutive patients aged 80 to 91 years (mean, 82.6 years; 86 men and 85 women) who underwent AVR at the Texas Heart Institute between 1975 and 1991. Seventy-seven patients had AVR only, and 94 patients had concomitant surgical procedures (coronary artery bypass graft surgery, 75 patients; mitral valve replacement, mitral valve repair, aneurysm repair, 19 patients). The overall 30-day early mortality was 17.5%. The early mortality was 5.2% for patients with AVR only and 27.7% for those with concomitant surgical procedures. Statistical analysis of 17 perioperative variables revealed that left ventricular ejection fraction of less than 45%, hypertension, congestive heart failure, angina, and concomitant surgical procedures were significant univariate predictors of early mortality. Multivariate analysis revealed that left ventricular ejection fraction of less than 45%, hypertension, and concomitant surgical procedures were independent predictors of operative mortality. Mean follow-up of survivors was 39 months. The overall actuarial survival at 1, 3, and 5 years was 90.8%, 84.2%, and 76.0%, respectively. CONCLUSIONS: These results show that AVR can be performed with acceptable operative risks in the elderly. This study further shows that isolated AVR can be done with low operative mortality and that the performance of concomitant surgical procedures exposes elderly patients to higher operative risks. PMID- 8222146 TI - Vasoreactivity of the radial artery. Comparison with the internal mammary and gastroepiploic arteries with implications for coronary artery surgery. AB - BACKGROUND: Recently, satisfactory results were obtained in a series of patients in whom the radial artery was used as a conduit for coronary artery bypass. However, spasm of this conduit was observed in 4% of patients. The aim of this study was to analyze the vasoreactive properties of the radial artery and to compare them with those of the internal mammary and the gastroepiploic arteries. METHODS AND RESULTS: Human radial (56 from 15 patients), internal mammary (77 from 20 patients), and gastroepiploic (41 from 12 patients) artery ring segments were mounted on a strain gauge in oxygenated, normothermic Krebs' solution at optimal resting tension. With KCl (100 mM) serving as the control, the dose response curves to norepinephrine, serotonin, and thromboxane A2 mimetic were obtained, permitting assessment of force of contraction and sensitivity. Functional endothelium was assessed with acetylcholine. Smooth muscle-dependent relaxation was assessed with sodium nitroprusside. The radial artery had stronger contractions to KCl than the other vessels. The radial and the gastroepiploic arteries with endothelium presented a higher contraction force than the internal mammary artery in response to norepinephrine and serotonin. The three vessels had equal sensitivities to norepinephrine and serotonin. The gastroepiploic artery had a lower sensitivity to thromboxane A2 mimetic than the two other vessels. CONCLUSIONS: This increased reactivity of the radial artery explains its propensity to spasm and emphasizes the need for antispastic drugs and platelet inhibitors when the radial artery is used for coronary artery bypass. PMID- 8222147 TI - Detection of intraluminal release of endothelium-derived relaxing factor from human saphenous veins. AB - BACKGROUND: Most prior physiological studies of the endothelium of intact saphenous veins have used organ chambers to assay abluminal release of vasoactive factors that affect smooth muscle contraction and relaxation. However, intraluminal release of vasoactive factors may have important implications in saphenous vein bypass graft patency. METHODS AND RESULTS: To study the basal and stimulated intraluminal release of endothelium-dependent relaxing factor (EDRF), 5-cm segments of human saphenous veins were cannulated and perfused in vitro with physiological salt solution. Vasoactive properties of the effluent were bioassayed on canine coronary artery smooth muscle. Effluent from saphenous veins produced vasodilation of 4.0 +/- 1.8% (+/- SEM) (n = 26) under basal conditions and 36.1 +/- 6.7% when stimulated with calcium ionophore, A23187 (n = 19, P < .05). Fourteen of the 26 vein segments showed no basal release of EDRF. Vasodilation produced by the effluent could be eliminated by mechanically removing the endothelium or by treatment with NG-monomethyl-L-arginine or NG nitro-L-arginine, two competitive inhibitors of nitric oxide synthesis from L arginine. Vasodilation was not influenced by indomethacin. CONCLUSIONS: Previous studies of EDRF from human saphenous veins were performed in organ chambers, which could detect only extraluminal EDRF release. This is the first demonstration of intraluminal release of EDRF from human saphenous veins. The variable and relatively poor basal release of EDRF into the lumen may account for early thrombogenicity of vein grafts and may contribute to late atherosclerosis. PMID- 8222148 TI - Comparison of phasic blood flow velocity characteristics of arterial and venous coronary artery bypass conduits. AB - BACKGROUND: Coronary artery bypass conduits derived from internal mammary arteries show relative resistance to atherosclerosis and significantly improved long-term patency compared with saphenous vein grafts. Atherothrombotic occlusion of venous conduits has previously been correlated with lower flow rates measured intraoperatively. To quantitate coronary bypass conduit flow velocity, we examined the phasic blood flow velocity patterns by intravascular Doppler spectral analysis in patients during cardiac catheterization to test the hypothesis that resting systolic and diastolic phasic blood flow velocity patterns differ significantly between arterial and venous bypass conduits. METHODS AND RESULTS: Spectral phasic blood flow velocity was measured using an intravascular Doppler-tipped angioplasty guidewire in the proximal, mid, and distal segments of 18 internal mammary artery conduits and 11 saphenous vein grafts in 27 patients at a mean of 4 years (range, 1 to 11) postoperatively. In situ internal mammary artery conduits demonstrated a gradual longitudinal transition in the phasic flow pattern from predominantly systolic velocity proximally (diastolic/systolic peak velocity ratio, 0.6 +/- 0.2) to predominantly diastolic velocity distally (diastolic/systolic peak velocity ratio, 1.4 +/- 0.3; P < .001). Saphenous vein graft flow velocity pattern, however, showed a consistently diastolic predominance, both proximally and distally (diastolic/systolic peak ratios, 1.4 +/- 0.6 and 1.5 +/- 0.7, respectively; P = NS). Mean flow velocities, total velocity integral, and calculated maximal shear rates were significantly higher in all segments of internal mammary arteries compared with values in saphenous vein grafts. CONCLUSIONS: Patterns of resting phasic blood flow, as well as mean velocity and total velocity integral, differ significantly between internal mammary artery and saphenous vein bypass conduits. These differences may have implications regarding blood-vessel wall interactions, the development of degenerative graft disease, and long-term conduit patency. PMID- 8222149 TI - Surgical treatment of discrete and tunnel subaortic stenosis. Late survival and risk of reoperation. AB - BACKGROUND: Although membranectomy, with or without septal myotomy or myectomy, has been the accepted method for treatment of fixed subaortic stenosis, controversies remain regarding operative methods and uncertainties regarding recurrence of subaortic obstruction and development of aortic insufficiency after repair. METHODS AND RESULTS: To determine late survival and risk of reoperation, we reviewed 169 patients who, between 1957 and 1992, had repair of discrete (n = 108) and tunnel (n = 61) subaortic stenosis. One hundred fifty-four patients had their primary operation for subaortic stenosis at the Mayo Clinic, and 15 patients had their primary operation elsewhere. Among patients having initial repair at our institution, membranectomy was performed in 52 patients (33.8%), membranectomy plus myotomy in 24 patients (15.6%), membranectomy plus myectomy in 71 patients (46.1%), myectomy plus a Konno-Rastan procedure in 5 patients (3.2%), and myectomy plus a modified Konno-Rastan procedure in 2 patients (1.3%). For all patients, early mortality was 4.7% (2.7% since 1961), and during follow-up extending to 29 years, there have been 16 late deaths. Twenty-six patients underwent a second or third operation for recurrent left ventricular outflow obstruction, including 11 of the 154 who had their primary operation at the Mayo Clinic (7.1%). Among the 21 patients who had a discrete lesion at initial repair and required reoperation, 19 (92%) were noted to have tunnel obstruction at reoperation. At late follow-up, the left ventricular outflow tract gradient was higher in patients with tunnel versus discrete obstruction (33 +/- 5 versus 24 +/ 17 mm Hg, P < .04), and 10-year survival was poorer (79% versus 91%, P < .02). Ten-year survival was worse in patients with tunnel lesions and associated cardiac anomalies versus those with isolated tunnel subaortic stenosis (64% versus 92%, P < .005). Some degree of aortic valve insufficiency was seen at late follow-up in 26% of patients, but in most cases this was mild. For patients with discrete subaortic stenosis, risk of late aortic insufficiency was 38.6% after isolated membranectomy, 27.8% after membranectomy and myotomy, and only 7.3% after membranectomy and myectomy (P < .004). Progression of aortic insufficiency requiring aortic valve replacement occurred in only 6 patients. CONCLUSIONS: Our results support the use of myectomy in conjunction with membranectomy for discrete subaortic stenosis. For restenosis and tunnel obstruction, more complete relief of subaortic stenosis by extended resection or a modified or classical Konno-Rastan procedure may improve late survival and reduce the incidence of recurrent subaortic stenosis and late aortic valve insufficiency. PMID- 8222150 TI - Aortic valve replacement in the elderly. Effect of gender and coronary artery disease on operative mortality. AB - BACKGROUND: Aortic valve replacement remains the treatment of choice for aortic valve disease, even in the extreme elderly who may present with advanced symptoms. Defining risk factors for short-term survival was the object of this study. METHODS AND RESULTS: This was a retrospective analysis of 717 patients at least 70 years of age who underwent aortic valve replacement alone or with coronary artery bypass graft between 1980 and 1992. Age range was 70 to 95 years, and mean age was 77 years; there were 529 septuagenarians (74%); 188 were octogenarians (26%); 326 were women (45%); and 386 patients (54%) had aortic valve replacement and coronary artery bypass graft. Atrial fibrillation/flutter or heart block was present in 16%, and 34% of patients were in New York Heart Association (NYHA) functional class IV. Aortic stenosis was present in 88%, and mechanical prostheses were used in 22% of patients. There were 47 deaths, giving an overall operative mortality of 6.6%, with 4.2% for aortic valve replacement and 8.8% for aortic valve replacement and coronary artery bypass graft (P = .01). The operative mortality for aortic valve replacement was 2.9% versus 10.3% for aortic valve replacement and coronary artery bypass graft in women (P = .006). The corresponding values for men were 5.6% and 7.4% (P = .31). Multivariate logistic regression showed coronary artery bypass graft and NYHA class IV to be significant predictors of operative mortality in women. The significant predictors in men were NYHA class IV, atrial fibrillation/flutter or heart block rhythm, and the use of mechanical prosthesis. Age was not a predictor of operative mortality in either sex. CONCLUSIONS: Aortic valve replacement carries an acceptable mortality rate in elderly patients. Female gender was a significant predictor of operative mortality in the concomitant coronary artery bypass graft group; however, gender was not a predictor of operative mortality in the isolated aortic valve replacement group. Advance stage of the disease process represented by NYHA class IV was a significant predictor of mortality for the whole group, stressing the need for earlier referral for surgery. PMID- 8222151 TI - Subaortic obstruction in double outlet right ventricles. Surgical considerations for anatomic repair. AB - BACKGROUND: Subaortic obstruction is one of the risk factors for anatomic repair of double outlet right ventricles (DORV). A comprehensive approach to such lesions has been developed in our institution since 1981. This retrospective work analyzes the results of this approach. METHODS AND RESULTS: Between January 1981 and September 1992, 30 patients aged 15 days to 15 years (mean, 44.8 months) underwent repair of a DORV associated with subaortic obstruction. Eighteen patients had a palliative procedure before complete repair. The ventricular septal defect (VSD) was subaortic in 15 patients, doubly committed in 1, noncommitted in 9, and subpulmonary in 5. The subaortic obstruction was a result of restrictive VSD in 29 patients and of double straddling of mitral and tricuspid valves once. The preoperative peak systolic pressure gradient between the left ventricle and the aorta (LV-Ao) was 68.7 +/- 23 mm Hg. Reconstruction of the left ventricular outflow tract comprised a ventral enlargement of the VSD in subaortic, doubly committed, and those subpulmonary VSDs scheduled for an arterial switch operation or a conal resection in noncommitted and other subpulmonary forms. Reconstruction of the right ventricular outflow tract included primary closure of the right ventricle in 12 patients, an infundibular patch in 9, a transannular patch in 4, and insertion of a right ventricular pulmonary valved conduit in 5. There were two early (6.6%) and two late (7.1%) deaths. Three patients required reoperation. A mean follow-up of 60.5 +/- 46.8 months was achieved in all the survivors. They were all in New York Heart Association class I or II, in sinus rhythm. At last follow-up, the mean LV-Ao gradient was 7.5 +/- 6.2 mm Hg, and LV function indices were within normal ranges. Actuarial survival and freedom from reoperation rates at 8 years were 86.6% and 87.0%, respectively. CONCLUSIONS: Surgical relief of subaortic obstruction in DORV has to be adapted to VSD location and spatial arrangement of atrioventricular valves and great vessels. PMID- 8222152 TI - Right ventricular decompression and left ventricular function in pulmonary atresia with intact ventricular septum. The influence of less extensive coronary anomalies. AB - BACKGROUND: Right ventricular decompression (RVD) may cause myocardial ischemia in patients with pulmonary atresia with intact ventricular septum and associated coronary artery abnormalities. Although we have previously shown that mortality is very high when two or more coronary arteries are obstructed, the effects of lesser degrees of coronary abnormalities are unknown. We therefore evaluated the effect of RVD on left ventricular (LV) function in those with less extensive coronary artery abnormalities. METHODS AND RESULTS: Preoperative cineangiograms demonstrated fistulas with or without one coronary artery stenosis in 12 of 24 patients aged 2 days to 33 months at the time of RVD. Preoperative and postoperative two-dimensional echocardiograms were analyzed for global and regional LV function. One infant with fistulas involving two coronary arteries and stenosis of the right coronary artery died from severe global LV dysfunction after RVD. Despite this, mean LV end-diastolic volume (66 +/- 17 mL/m2) and mean LV ejection fraction (60 +/- 9%) were similar in patients with and without coronary artery abnormalities before and after RVD. Before RVD, regional LV dysfunction was seen in 8 of 132 (6%) regions in those with coronary artery abnormalities and in 3 of 132 (2%) in those without coronary artery abnormalities. After RVD, there were 16 of 132 (12%) abnormal regions in those with coronary artery abnormalities and 1 of 132 (< 1%) in those without coronary artery abnormalities. In regions with normal wall motion before RVD, the presence of coronary artery abnormalities was related to regional LV dysfunction after RVD (P < .001). CONCLUSIONS: Regional LV dysfunction was rare in patients without coronary artery abnormalities. In those with less extensive coronary artery abnormalities not involving obstruction to multiple coronary arteries, regional LV dysfunction was common before and increased after RVD, but severe global LV dysfunction was unusual. PMID- 8222153 TI - Restoration of the pulmonary valve reduces right ventricular volume overload after previous repair of tetralogy of Fallot. AB - BACKGROUND: Severe pulmonary regurgitation (PR) and associated right ventricular (RV) dilatation are late complications of surgical repair of tetralogy of Fallot (TOF). For the past several years, we have restored pulmonary valve competence with the exclusive use of cryopreserved allografts. METHODS AND RESULTS: Sixteen patients with symptoms of diminished exercise tolerance and echocardiographic evidence of progressive PR with severe RV dilatation underwent placement of allografts in the RV outflow tract at a median age of 12 years (10 years after TOF repair). Abnormal exercise tolerance tests were documented in 10 patients. Additional surgical procedures included pulmonary artery augmentation (n = 6), closure of residual left to right shunts (n = 3), and subendocardial resection for monomorphic ventricular tachycardia (n = 1). Six patients had either preoperative or postoperative balloon dilations of pulmonary artery stenoses. All patients had symptomatic improvement after allograft insertion. At a mean follow up of 26.4 +/- 3.4 months, the severity of PR improved in all but one patient. In 12 patients (group 1), conduit regurgitation was either trace (n = 11) or mild (n = 1). Four patients (group 2) had moderate conduit regurgitation. In a retrospective analysis, pulmonary artery diameters and cross-sectional areas were significantly smaller in the group 2 patients compared with the group 1 patients. With the exception of one patient, RV end-diastolic diameter (RVEDD/BSA) fell after allograft insertion in each patient (P < .01). The reduction in RVEDD/BSA was significantly greater in group 1 than in group 2 (31.8 +/- 3.4% versus 21.4 +/- 11.0%, P < .05). CONCLUSIONS: Thus, restoration of the pulmonary valve with cryopreserved allografts improved exercise tolerance and diminished RV volume overload in patients with severe PR after previous repair of TOF. Optimal results were achieved in patients who did not have significant residual pulmonary artery distortion. PMID- 8222154 TI - Neonatal coarctation repair. Influence of technique on late results. AB - BACKGROUND: Coarctation repair in the neonate (< or = 28 days) is associated with higher mortality and increased incidence of restenosis compared with older infants. It has been suggested that resection of pericoarctation ductal tissue may reduce this risk of restenosis. METHODS AND RESULTS: To further clarify these issues, we reviewed our experience with 111 consecutive neonates undergoing primary repair between 1973 and 1991. Hospital mortality was 14.4% (16 of 111) and was not significantly different for the type of repair:resection and end-to end anastomosis (RETE) 10.7% (6 of 56), subclavian flap angioplasty (SFA) 16.7% (6 of 36), and patch angioplasty (PA) 16.7% (3 of 18). Associated complex cardiac pathology was associated with higher operative risk: 25% (10 of 40) versus 8.4% (6 of 71) (P = .02). Median follow-up of 4.2 years (range, 0.1 to 18.5 years) was 99% complete. Late mortality was 13.6% (13 of 95), of which 92% occurred within 1 year of repair. Twenty percent (19 of 95) needed reintervention for restenosis, RETE 16% (8 of 50), SFA 13% (4 of 30), and PA 47% (7 of 15) (P = .02). Of these, 84.2% (16 of 19) required reintervention within 1 year of repair. Freedom from reintervention 1 and 8 years after operation was 80 +/- 4% and 77 +/- 5%, respectively. Actuarial survival 8 years after operation was 73 +/- 4%; for simple coarctation, this was 90 +/- 4%. By multivariate analysis, survival was negatively influenced only by presence of associated cardiac pathology (P = .002) and reintervention only by patch angioplasty technique of repair (P = .007). CONCLUSIONS: In the neonate, resection of coarctation (RETE) does not diminish the risk for reintervention compared with SFA. The risk for both late death and recurrent coarctation are highest within the first year after repair, and follow up should be particularly vigilant during this period. PMID- 8222155 TI - Intraoperative device closure of ventricular septal defects. AB - BACKGROUND: Surgical repair of muscular ventricular septal defects (VSDs) has been associated with significant morbidity and mortality when the defects are multiple, associated with complex cardiac lesions, or requiring left ventriculotomy. Transcatheter VSD closure may be difficult in patients weighing less than 7 kg or those with limited vascular access. We attempted intraoperative device closure of muscular VSDs during surgical repair of associated lesions. METHODS AND RESULTS: We considered intraoperative umbrella closure in 10 patients, 8 with additional complex heart lesions. "Swiss cheese" defects were present in 4. Median patient age was 4.7 months. Device closure was not attempted in 1 patient due to an inadequate septal rim. Nine umbrellas, 12 to 40 mm in diameter, were positioned to straddle the septum in all 9 patients. There were 3 early deaths, 2 in patients who were moribund preoperatively. The third was due to severe ventricular dysfunction. Among the 6 survivors, 3 had a Qp:Qs < or = 2 and right ventricular or pulmonary artery pressure less than half systemic. These patients had no further intervention. The other 3 had a Qp:Qs > 2. One died unexpectedly 5 weeks after cardiac surgery. The other 2 had subsequent closure of residual VSDs. All 5 survivors are well at follow-up of 8 to 25 months. CONCLUSIONS: Intraoperative umbrella VSD closure appears less successful than VSD closure in the catheterization laboratory. In selected patients, particularly very small infants, intraoperative device placement may be advantageous. Completeness of closure is difficult to assess intraoperatively. PMID- 8222156 TI - Intraoperative and percutaneous stenting of congenital pulmonary artery and vein stenosis. AB - BACKGROUND: Conventional surgical or balloon dilation therapy for pulmonary artery or vein stenosis has been unsatisfactory in many patients. Balloon expandable stents offer a new form of treatment for these vascular stenoses and can be implanted percutaneously or intraoperatively. METHODS AND RESULTS: Between July 1991 and October 1992, 20 balloon-expandable Palmaz stents (Johnson & Johnson) were implanted in 16 children at median age and weight of 3.0 years and 12.8 kg, respectively. Stent implantation was performed intraoperatively (n = 15) if the patient was less than 1 year of age or less than 10 kg in weight, in cases where limited vascular access precluded percutaneous implantation, or as an adjunct to other intracardiac surgery. Otherwise, percutaneous stenting was performed (n = 5). Vessels were tested for distensibility by dilation with balloon catheters or vascular sounds. Stents were implanted using angioplasty catheter balloons chosen to achieve desired vessel diameter and inflated to 4 to 17 atm. Acute hemodynamic and cineangiographic studies were performed in all patients immediately after the procedure to 2 months after stenting. After pulmonary artery stent implantation, mean pulmonary artery diameter increased from 5.6 to 11.5 mm (P = .001), with a decrease in mean systolic pressure gradients from 43 to 8.0 mm Hg (P = .005). Follow-up cardiac catheterization (mean, 8.7 months) in 3 patients revealed no restenosis, thrombosis, or aneurysm formation. In patients in whom pulmonary vein stents were implanted, mean pressure gradients fell from 11 to 0.3 mm Hg (P = .03), and mean pulmonary capillary wedge pressure fell from 17 to 6.3 mm Hg (P = .03) immediately after stenting. At 2- to 6-month follow-up, cardiac catheterization documented restenosis within the stent in 2 of 3 patients. The third patient died 2 months after stenting from presumed vein reocclusion. CONCLUSIONS: When implanted intraoperatively or percutaneously, balloon-expandable endovascular stents have been efficacious in the treatment of pulmonary artery stenosis. Longer follow-up will be necessary to document the long-term effectiveness of pulmonary artery stenting. Preliminary data suggest that early restenosis is common after pulmonary vein stenting. The intraoperative approach extends stenting therapy to smaller children and to patients who have limited percutaneous access. PMID- 8222157 TI - Improved survival following pediatric cardiac transplantation in high-risk patients. AB - BACKGROUND: Preoperative hemodynamic support, complex congenital heart disease, and elevated pulmonary vascular resistance present particular challenges for pediatric heart transplantation. This study was performed to identify preoperative factors that influence survival after pediatric heart transplantation over two eras of pediatric heart transplant experience. METHODS AND RESULTS: We retrospectively analyzed demographic, clinical, and hemodynamic data from 67 pediatric patients who underwent heart transplantation between February 1982 and June 1992 and compared survival between two eras (early experience versus late experience). During the early experience (group 1: February 1982 to August 1989), univariate analysis identified congenital heart disease, pretransplant extracorporeal membrane oxygenator (ECMO) support, inotropic and/or ventilatory support (UNOS status I), elevated transpulmonary gradient (at least 15 mm Hg), and elevated pulmonary vascular resistance index (at least 4 Wood units.m2) as preoperative risk factors for early death after pediatric heart transplantation. However, in the late experience (group 2: September 1989 to June 1992), the only risk factor for premature death by univariate analysis was elevated transpulmonary gradient. By multivariate analysis, elevated transpulmonary gradient was the only risk factor for our early, late, and entire experiences. One-year survival after transplantation for congenital heart disease was improved from 46% in group 1 to 73% in group 2 (P < .05). In group 1, only one patient (25%) with pretransplant ECMO support survived 1 year, whereas 66% (four of six) survived more than 1 year in group 2. CONCLUSIONS: Although elevated transpulmonary gradient continues to be a significant risk factor for pediatric heart transplantation, candidates with congenital heart disease, UNOS status I, and pretransplant ECMO support now can be successfully transplanted with reasonable hope for extended survival. PMID- 8222158 TI - Decreasing incidence of coronary disease in pediatric cardiac transplant recipients using increased immunosuppression. AB - BACKGROUND: Coronary artery disease (CAD) is a limiting factor to long-term survival in cardiac transplant recipients, affecting from 30% to 50% of patients by 4 years after surgery. Can the incidence of CAD be lowered with augmentation of immunosuppression? METHODS AND RESULTS: We compared the incidence of CAD in our pediatric transplant population with nine potential risk factors, including immunosuppressive regimen. The study group consisted of 55 patients who survived more than 1 year (or to first angiogram) or had autopsies. Coronary angiograms were performed yearly and compared sequentially. The mean follow-up of 55 patients was 36 months. Mean age was 10.3 +/- 6 years (range, 4 months to 18 years). Thirteen patients received double immunosuppression with cyclosporine and prednisone, and 42 received triple therapy with cyclosporine, prednisone, and azathioprine. Significant CAD occurred in 10 grafts (6 deaths and 3 retransplants). Cause for graft loss in 6 patients with CAD was acute rejection. CAD was detected by angiogram in only 2 patients. Nine of 10 patients received double therapy (P < .001). There was no difference in mean follow-up between immunosuppression groups. There was a higher rejection frequency for double therapy (0.19 +/- 0.16 rejections per patient month) compared with triple therapy (0.07 +/- 0.11). Ten patients were rejection free in the triple therapy group. CONCLUSIONS: We experienced a significant decrease in the incidence of CAD in our pediatric cardiac transplant recipients using increased immunosuppressive therapy. Type of immunosuppressive regimen (double) and rejection frequency were independent predictors for CAD by multivariate analysis. PMID- 8222159 TI - Neoplastic disorders after pediatric heart transplantation. AB - BACKGROUND: Because of their life-long requirement for immunosuppressive therapy, neoplastic disorders could represent a significant threat to long-term survival in infants and children after heart transplantation. This study determined the incidence and clinical spectrum of neoplastic disorders in 80 pediatric patients who underwent heart transplantation between 1974 and 1992. METHODS AND RESULTS: Follow-up ranged from 6 to 189 months (mean, 50.0 months). Tumors occurred in 10 patients (12.5%). Time to detection ranged from 3.3 to 139.2 months (mean, 52.7 months). Tumor incidence was greatest in 9 patients transplanted before the cyclosporine era (44%) compared with the subsequent 71 patients (8.5%, P < .05). There was no increase in risk related to sex, age, underlying disease, or blood type; however, patients with tumors received higher initial doses of cyclosporine and prednisone and had more rejection episodes in the first 3 months (P < .05). There was an increased risk associated with anti-thymocyte globulin (33%, P < .05) but not with OKT3 (6%, P = NS). There were eight lymphoproliferative disorders (four B-cell, one T-cell, three not determined) and one hepatocellular and one squamous cell carcinomas. Six cases of lymphoproliferative disorder had in situ evidence of Epstein-Barr virus. Patients were treated by reducing immunosuppression (7), radiotherapy (2), and chemotherapy (1). There were five deaths: two tumor related and the others due to rejection, renal failure, and infection. Of 5 survivors, 1 had tumor recurrence 4 years after diagnosis, and 4 are disease free. CONCLUSIONS: Tumors represent a small but serious long-term risk to pediatric heart transplant recipients. The incidence in children transplanted in the cyclosporine era is similar to that in adults, and the majority of tumors are lymphoproliferative disorders that often regress by reducing immunosuppression. PMID- 8222160 TI - Exercise performance after pediatric heart transplantation. AB - BACKGROUND: Children awaiting heart transplantation have severe limitations in their ability to exercise. The purpose of this study was to assess exercise capacity after pediatric heart transplantation and to identify factors influencing exercise performance. METHODS AND RESULTS: Progressive cycle ergometry testing was performed in 31 patients at 1.3 +/- 0.8 years after transplantation, and in 16 patients, follow-up studies were performed at 3.3 +/- 1.3 years after transplantation. Maximum work load (Wmax), peak oxygen consumption (VO2), and maximum heart rate (HRmax) were measured. Exercise capacity was defined as normal if Wmax was > or = 75% of predicted values and decreased if Wmax was < 75% of predicted values. Differences in age at transplantation, sex, diagnosis, duration of heart failure, New York Heart Association class before transplantation, resting cardiac index, body mass index, and rejections per patient month were compared between patients with normal and decreased exercise capacity. At initial study, Wmax was 62 +/- 38 W or 61% of that predicted, peak VO2 was 20 +/- 6 mL.kg-1 x min-1 (63% of that predicted), and HRmax was 136 +/- 22 beats per minute (66% of that predicted) for all 31 patients. Six patients had normal exercise capacity, and 25 patients had decreased exercise capacity. Peak VO2 was significantly higher in the normal versus the decreased exercise capacity patients (26 +/- 5 vs 19 +/- 5 mg.kg-1 x min-1). The mean age at transplantation was significantly less in patients with normal exercise capacity: 8.2 +/- 4.6 versus 12.5 +/- 3.6 years for patients with decreased capacity. On follow-up study, no significant differences in Wmax, peak VO2, or HRmax were found from the initial test. CONCLUSIONS: Similar to results obtained in adult patients, exercise capacity was decreased but stable in pediatric patients after heart transplantation. PMID- 8222161 TI - Mechanism of outflow tract obstruction causing failed mitral valve repair. Anterior displacement of leaflet coaptation. AB - BACKGROUND: Systolic anterior motion of the mitral valve causing left ventricular outflow tract obstruction occurs in 1% to 2% of patients having mitral valve repair, in some cases requiring further surgery to relieve the obstruction, but the mechanism and the geometry involved are not certain. METHODS AND RESULTS: We studied 14 patients who developed systolic anterior motion and left ventricular outflow tract obstruction, all after posterior leaflet resection and annuloplasty, in whom a second repair eliminated systolic anterior motion by complete (n = 6) or partial (n = 8) ring removal. Intraoperative transesophageal echocardiography was recorded before pump, after failed repair during left ventricular outflow tract obstruction, and after a corrective second pump run to relieve the systolic anterior motion. Systolic anterior motion occurred when the mitral valve coaptation to septum distance was reduced (before, 26.5 +/- 4.3; during systolic anterior motion, 17.4 +/- 4.4 versus after second pump, 23.4 +/- 6.9 mm) and the mitral valve coaptation to posterior mitral annulus distance was greater (before, 18.9 +/- 3.4; during systolic anterior motion, 22.2 +/- 4.6 versus after second pump, 17.4 +/- 3.6 mm), both P < .01. Comparing dimensions before pump, during systolic anterior motion, and after the second pump, there were no differences in left ventricular cavity diameter in systole or diastole, the septum to posterior annulus distance, or the angle between the aortic and mitral annular planes. CONCLUSIONS: After mitral repair, left ventricular outflow tract obstruction occurs when the mitral coaptation line is displaced anteriorly. When systolic anterior motion occurs, reduction of the amount of annuloplasty or use of the posterior leaflet sliding procedure may eliminate this problem. Understanding the geometry of this phenomenon may facilitate preoperative echo selection of high-risk patients (those with large redundant posterior leaflets and relatively normal ventricular size) and modification of surgical technique to avoid the problem of outflow tract obstruction after mitral valve repair. PMID- 8222162 TI - Coronary artery endothelial interleukin-1 beta mediates enhanced fibronectin production related to post-cardiac transplant arteriopathy in piglets. AB - BACKGROUND: Graft coronary arteriopathy has become the major complication observed in the late follow-up of cardiac transplant patients. We investigated this process experimentally in piglets after a heterotopic cardiac transplant and observed early changes in donor coronary arteries compatible with an immune inflammatory process, ie, there is increased expression of interleukin-1 beta (IL 1 beta), fibronectin, and activated lymphocytes associated with intimal thickening. METHODS AND RESULTS: In this study, we cultured porcine coronary artery endothelial cells from host and donor hearts and found similarities in morphology and uptake of acetylated low-density lipoprotein (LDL). As well, host and donor cells showed similar patterns of growth, protein, and glycosaminoglycan synthesis. Endothelial cell fibronectin synthesis was determined after radiolabeling with [35S]-methionine in serum-free medium, gelatin-sepharose extraction of the culture medium and resolution on 5% SDS-PAGE. Donor coronary artery endothelial cell fibronectin synthesis was up to fivefold higher than that of host but was not associated with comparable increased levels of fibronectin mRNA. IL-1 beta appeared to mediate this enhanced fibronectin production, since the IL-1 receptor antagonist caused a 50% decrease in this feature, a change not observed in host cells. Furthermore, donor endothelial cells produced twice the amount of IL-1 beta compared with host cells as judged by immunoprecipitation. CONCLUSIONS: Increased donor coronary artery endothelial cell fibronectin appears to be regulated at least partly by an autocrine mechanism involving IL-1 beta, and fibronectin may mediate lymphocyte trafficking and smooth muscle cell migration related to graft arteriopathy. PMID- 8222163 TI - Flow-mediated vasodilator response to tachycardia of epicardial coronary arteries is preserved in heart transplant recipients. AB - BACKGROUND: Increasing blood flow through conduit arteries induces vasodilation through endothelium-dependent mechanisms. In humans, flow-mediated dilation of angiographically normal epicardial coronary arteries has been observed during tachycardia, this response being impaired in the presence of atherosclerosis. METHODS AND RESULTS: To evaluate whether the endothelium-dependent physiological vasodilatory response of epicardial coronary arteries to tachycardia is preserved in heart transplant recipients, 22 patients with angiographically smooth coronary arteries were studied with quantitative angiography. A total of 14 patients had undergone cardiac transplantation more than 1 year (mean, 28 +/- 18 months) before the study, and 8 patients were nontransplant patients with atypical chest pain and normal exercise tests (control group). Angiograms of the left coronary artery were obtained on 35-mm cinefilms at 3-minute intervals in basal conditions, during pacing-induced tachycardia (150 beats per minute), and after intracoronary injection of 1.5 mg of isosorbide dinitrate. During tachycardia, the mean luminal diameter of the midsegment of the left anterior descending coronary artery increased by 8.9 +/- 6.1% from 2.64 +/- 0.56 to 2.88 +/- 0.62 mm (P < .001) in transplant recipients and by 7.5 +/- 5.0% from 2.37 +/- 0.54 to 2.53 +/- 0.50 mm (P < .025) in the control group (transplant vs control patients, NS). A further coronary dilation was observed in all patients after isosorbide dinitrate, up to 124.8 +/- 8.1% of basal lumen diameter in transplant recipients and up to 129.1 +/- 16.1% of basal diameter in the control group. CONCLUSIONS: The vasodilator response of epicardial coronary arteries to tachycardia is preserved in heart transplant recipients. This suggests that the functional response of the endothelium to an increase in coronary blood flow remains normal in these patients. PMID- 8222164 TI - Coronary collateral circulation in the transplanted heart. AB - BACKGROUND: The concept that coronary collateral (CC) circulation can develop in cardiac transplant recipients (CTR) is controversial. Indeed, a decreased occurrence of CC in CTR has been previously reported. METHODS AND RESULTS: We reviewed 102 coronary angiograms in 73 CTR to evaluate the presence and significance of CC in denervated human hearts. These angiograms were compared with angiograms of 70 nontransplanted patients. Twenty-six CTR who had undergone at least two coronary angiograms, thus allowing comparative evaluations, form the basis for this study. Angiograms were analyzed using a modification of CC classification (Rentrop et al) from grade 0 for complete absence to grade 4 representing mature collateral with clear filling of epicardial vessels. Coronary artery disease was classified according to the scheme reported by Gao et al. For the purpose of this study, all type A lesions were grouped as large vessel disease (LVD), and types B1, B2, and C were collectively grouped as small vessel (epicardial) disease (SVD). The presence of CC circulation in all 73 CTR was grade 0, 7 (10%); grade 1, 41 (56%); grade 2, 33 (45%); grade 3, 30 (41%); and grade 4, 5 (7%). However, in control nontransplanted subjects grades 0, 1, 2, 3, and 4 were found in 47 (67%), 22 (31%), 4 (6%), 11 (15%), and 16 (20%) patients, respectively. The presence of mature collaterals (grade 4) in both groups were associated with type A lesions and was frequent in nontransplanted hearts. In contrast, grade 2 and grade 3 vascular channels probably representing CC with "myocardial blush" was more frequent in CTR and was mostly associated with small vessel coronary arteriopathy. CONCLUSIONS: These results suggest the presence of atypical CC in patients with cardiac allograft arteriopathy. It is speculated that this atypical form of CC with "blush pattern" may represent an angiogenic response to microvascular ischemia due to allograft coronary arteriopathy. PMID- 8222165 TI - Brain death-induced cardiac contractile dysfunction and long-term cardiac preservation. Rat heart studies of the effects of hypophysectomy. AB - BACKGROUND: Ischemic brain death induces cardiac contractile dysfunction, which may exclude the heart as a donor organ for transplantation; the mechanism is unknown. Since cerebral ischemia might alter pituitary function, we investigated the influence of hypophysectomy on basal contractile function, brain death induced contractile malfunction, and the tolerance of the heart to hypothermic ischemic storage. METHODS AND RESULTS: Rats were hypophysectomized and maintained for 5 days; during this time, left ventricular developed pressure (LVDP) fell to 70% of its control value (92 +/- 8 versus 132 +/- 6 mm Hg, P < .05). Diastolic function of isolated blood-perfused hearts 5 days after hypophysectomy was severely impaired (left ventricular volume at 12 mm Hg of end-diastolic pressure was 141 +/- 20 versus 250 +/- 30 microL in sham-operated control rats; P < .05). Brain death in nonhypophysectomized rats resulted in a transient increase in mean arterial pressure (from 112 +/- 4 to 180 +/- 7 mm Hg within the first 30 seconds) followed by a rapid decline to less than 50% of the control value (54 +/- 3 mm Hg after 5 minutes, P < .05). Changes in cardiac function were comparable (cardiac index fell from 34 +/- 2 to 17 +/- 1 mL/min per 100 g body weight, and stroke volume index fell from 82 +/- 5 to 41 +/- 4 microL per beat per 100 g body weight within 10 minutes; P < .05). Brain death in hypophysectomized rats resulted in a similar (but attenuated) biphasic response with arterial pressure (already reduced to 82 +/- 9 mm Hg as a consequence of hypophysectomy) transiently increasing to 144 +/- 9 mm Hg after 30 seconds and then falling to 52 +/- 5 mm Hg by 5 minutes. Finally, we assessed the effects of coincident hypophysectomy and brain death on the ability of the heart to recover from 6 hours of hypothermic (4 degrees C) ischemic storage. Brain death alone had no effect on the postischemic recovery of LVDP (133 +/- 14 versus 129 +/- 12 mm Hg at 8 mm Hg of end-diastolic pressure in the non-brain death group). LVDP in hearts from hypophysectomized rats with brain death recovered to only 84 +/- 13 mm Hg; however, this was virtually identical to the LVDP in hearts from hypophysectomized rats that had not been subjected to brain death (83 +/- 5 mm Hg) or had not even been exposed to hypothermic ischemia (83 +/- 10 mm Hg). CONCLUSIONS: Hypophysectomy induces a deterioration of cardiac function that becomes apparent after 2 days. However, it does not exacerbate the cardiac dysfunction induced by brain death. Hearts from hypophysectomized animals, with or without brain death, recovered less well after prolonged hypothermic storage; nevertheless, the hearts recovered to preischemic levels, indicating that, although hypophysectomy impairs cardiac contractile function, it does not adversely influence the tolerance of the heart to hypothermic ischemia. Pituitary function may be an important factor in determining cardiac function without influencing resistance to ischemia or responses to brain injury. PMID- 8222166 TI - Cyclosporine-induced hypertension. Efficacy of omega-3 fatty acids in patients after cardiac transplantation. AB - BACKGROUND: Cyclosporine-induced hypertension may be related to vasoconstriction of the afferent arterioles in the glomeruli caused by changes in the prostaglandin profile. omega-3 Fatty acids have demonstrated vasodilatory properties related to a favorable effect in the prostaglandin profile. The purpose of this study was to evaluate the antihypertensive effects of oral supplementation with omega-3 fatty acids in cyclosporine-treated cardiac transplant recipients. METHODS AND RESULTS: The study consisted of 20 orthotopic cardiac transplant recipients with hypertension who were prospectively randomized in a double-blind fashion to receive either omega-6 fatty acids (placebo group, n = 10) or omega-3 fatty acids (treatment group, n = 10). Blood pressure, systemic hemodynamics, two-dimensional guided M-mode and Doppler echocardiography, and laboratory values (serum creatinine, lipid profile) were recorded at baseline and at 12 weeks. The treatment group demonstrated a significant reduction in mean arterial pressure (120 +/- 7 versus 102 +/- 7 mm Hg; P = .0001) associated with a decrease in systemic vascular resistance (2107 +/- 45 versus 1426 +/- 60 dynes.sec.cm-5; P = .0001). No changes in indexes of left ventricular structure and function occurred, except for a modest decrease in deceleration time (211 +/- 10 versus 182 +/- 12 milliseconds; P = .05), an index of left ventricular diastolic function. CONCLUSIONS: omega-3 Fatty acids (3 g/d) reduce blood pressure by decreasing systemic vascular resistance and, therefore, can be used as an adjuvant for the treatment of hypertension in cyclosporine-treated cardiac transplant recipients. Their vasodilatory effect may be related to a beneficial change in the prostaglandin profile. PMID- 8222167 TI - Captopril reduces graft coronary artery disease in a rat heterotopic transplant model. AB - BACKGROUND: Transplant coronary artery disease (CAD) is characterized by severe myointimal proliferation causing vascular stenosis. Nontransplant vascular injury models have shown that angiotensin converting enzyme (ACE) inhibitors reduce myointimal proliferation and preserve lumen integrity. We examined the effect of the ACE inhibitor captopril on graft CAD in a Lewis to F344 rat heterotopic cardiac transplantation model. METHODS AND RESULTS: Twenty-five control rats (group 1) were observed without captopril administration after heterotopic cardiac transplantation, and the other 19 rats (group 2) were administered captopril (50 mg/kg per day in drinking water) after heart transplantation. Graft survival 3 months after transplantation was significantly (P < .02) higher in group 2 (18 of 19, 95%) than that in group 1 (16 of 25, 64%). Cellular rejection grades of the heart allografts were significantly higher in group 1 than those in group 2 both at 3 months (grades, 2.5 +/- 0.4 vs 1.3 +/- 0.7; P < .01) and 6 months (grades, 2.4 +/- 0.9 vs 0.8 +/- 0.5; P < .05) after transplantation. The grades of graft CAD (vascular rejection) were also significantly higher in group 1 than those in group 2 both at 3 months (grades, 2.3 +/- 0.7 vs 0.9 +/- 0.9; P < .05) and 6 months (grades, 3.0 +/- 0.9 vs 0.9 +/- 0.3; P < .01) after transplantation. The cardiac allographs in group 2 showed minimal intimal proliferation, intact elastic laminae, and reduced smooth muscle cell proliferation. CONCLUSIONS: These results suggest that the ACE inhibitor captopril may be effective in prevention of accelerated graft CAD. PMID- 8222168 TI - Novel preservation solution permits 24-hour preservation in rat and baboon cardiac transplant models. AB - BACKGROUND: Cardiac preservation for transplantation disrupts normal vascular homeostatic mechanisms. Hypoxia and reoxygenation increase endothelial cell permeability, induce procoagulant activity, and alter endothelial cell/leukocyte interactions, with a parallel reduction in endothelial cAMP and nitric oxide levels. Because hypoxia/reoxygenation simulates a significant component of the global ischemia/reperfusion of cardiac transplantation, we hypothesized that preservation failure may be related to these perturbations. This work focuses on repleting the intercellular/intracellular second messengers nitric oxide/cGMP and cAMP in the donor heart to enhance cardiac preservation for transplantation. METHODS AND RESULTS: A heterotopic rat heart transplant model was used to compare lactated Ringer's (LR) and University of Wisconsin (UW) solutions to a novel storage solution (Columbia University solution, CU), which contains a cAMP analogue (dibutyryl cAMP) and nitroglycerin (to enhance nitric oxide-related mechanisms). By 24 to 28 hours of preservation, no LR hearts survived (n = 9), 35% of UW hearts survived (n = 20), and 88% of CU-preserved hearts survived (n = 8; P < .05) versus LR or UW). The viability of preserved hearts was explored by determining whether CU preservation enabled myocytes to maintain resting membrane potentials and preserve their ability to generate an action potential in response to a field stimulus. Of 24 sites explored with a microelectrode in UW-preserved hearts, only 4% were able to generate an action potential, compared with 75% of 36 sites in CU-preserved hearts (P < .001), with corresponding preservation of resting membrane potential in the CU-preserved hearts (-13 mV for UW versus -54 mV for CU, P < .001). Orthotopic baboon cardiac transplantation performed after 24-hour simple hypothermic preservation demonstrated that no UW-preserved heart (n = 4) survived the perioperative period; in contrast, four of five hearts stored for 24 hours in CU solution sustained the recipient with minimal inotropic support, and two animals survived long-term. CONCLUSIONS: Sustaining higher levels of intercellular/intracellular second messengers cAMP and nitric oxide/cGMP provides a new approach to enhancing cardiac preservation. PMID- 8222169 TI - Early effects of right latissimus dorsi cardiomyoplasty on left ventricular function. AB - BACKGROUND: We hypothesized that left ventricular function could be improved with cardiomyoplasty using the right latissimus dorsi. METHODS AND RESULTS: Five dogs underwent cardiomyoplasty using the right latissimus dorsi. Left ventricular volume and pressure were measured using sonomicrometry and a micromanometer catheter, respectively. Pressure volume loops were recorded with the muscle stimulated at 1:2 and with transient caval occlusion. During stimulated beats, there were significant increases in stroke work (13.90 +/- 4.49 vs 9.78 +/- 3.81 g/m, P < .01), preload recruitable stroke work (0.766 +/- 0.110 vs 0.594 +/- 0.207 g.m-1 x m-3, P < .05), and stroke volume (15 +/- 4 vs 10 +/- 3 mL, P < .05) when compared with unstimulated beats. There were no changes in diastolic filling. This operation was done in 11 patients, with no operative deaths. Six weeks after surgery, resting left ventricular ejection fraction (LVEF) increased from 25 +/- 1.6% to 35 +/- 3% (P < .05), and left ventricular end-diastolic volume (LVEDV) decreased from 365 +/- 18 to 307 +/- 24 mL, (P < .05). Nine patients were alive at 6 months. Preoperative and 6-month LVEF and LVEDV for those 9 patients were 26 +/- 2% and 29 +/- 2% (P = NS) and 316 +/- 23 and 261 +/- 22 mL (P < .05), respectively. CONCLUSIONS: Long-term studies are needed to determine if these changes will improve patient survival. PMID- 8222170 TI - Left ventricular outflow tract obstruction after mitral valve repair. Results of the sliding leaflet technique. AB - BACKGROUND: Left ventricular outflow tract obstruction (LVOTO) occurs in 4% to 5% of patients after prosthetic ring mitral valve repair. Major anatomic factors incriminated in the genesis of LVOTO include degenerative mitral valve insufficiency with excess leaflet tissue, nondilated left ventricular cavity, and narrow mitro-aortic angle. We have previously reported a 14% incidence of LVOTO after prosthetic ring mitral valve repair in this high-risk group of patients. Serial echo Doppler studies demonstrated an overlapping and/or inversion of the left ventricular functional compartments generating systolic anterior motion of the posterior leaflet and paradoxical opening (eversion) of the anterior leaflet. In an attempt to eliminate LVOTO after mitral valve repair, a new surgical procedure was developed in 1988 by Carpentier: the sliding leaflet technique, which reduces the height of the posterior leaflet. The purpose of this study was to analyze the results of the new technique in terms of the occurrence of LVOTO: METHODS AND RESULTS: Eighty-two patients undergoing prosthetic ring mitral valve repair between 1988 and 1991 and identified as high risk for LVOTO were operated on using the sliding leaflet technique. There were 52 men and 30 women. Ages ranged from 28 to 75 years. The surgical techniques used included prosthetic ring annuloplasty (n = 82), leaflet resection (n = 82), chordal shortening or transposition (n = 36), and other (n = 19). Intraoperative and/or immediate postoperative echo Doppler studies were obtained in all cases. Two patients (2.4%) died, and 2 (2.4%) required reoperation. Nonsignificant LVOTO was identified in 2 cases (2.4%), in whom instantaneous maximal subaortic gradients were 20 and 18 mm Hg, respectively. CONCLUSIONS: This study was not done on a concomitant series of patients but on patients with the same type of pathology. It demonstrates that (1) the sliding leaflet technique eliminates significant LVOTO in the high-risk patients; (2) the sliding leaflet technique is associated with a low mortality; and (3) no reoperations for mitral insufficiency were required in this series. PMID- 8222171 TI - Late effects of cardiomyoplasty on left ventricular mechanics and diastolic filling. AB - BACKGROUND: Dynamic cardiomyoplasty is a relatively new surgical procedure proposed for treatment of severe myocardial failure. Limited clinical experience suggests that this procedure reverses congestive heart failure, improving the long-term survival. METHODS AND RESULTS: In this investigation, the late effects (16.5 +/- 4.8 months after surgery) of cardiomyoplasty on left ventricular mechanics and diastolic filling were studied in eight male patients (mean age, 45 +/- 7 years). The investigation was performed with the myostimulator on and off (24 +/- 1 hours off) using a microtip catheter to obtain the left ventricular pressures simultaneously with Doppler (inflow velocities) and M-mode and two dimensional echocardiographic recordings. Statistical comparisons were made with Student's t test for paired data. Dynamic cardiomyoplasty increased the maximal elastance (17 +/- 1 versus 21 +/- 1 mm Hg/cm, P = .006) and decreased the systolic wall stress (253 +/- 17 versus 190 +/- 12 g/cm2, P = .029). Left ventricular end-diastolic pressure decreased (27.1 +/- 2.8 versus 17.6 +/- 1.7 mm Hg, P = .019), as did end-diastolic circumferential wall stress (69 +/- 8 versus 37 +/- 5 g/cm2, P = .002). The chamber and muscle stiffnesses decreased (120 +/- 31 versus 52 +/- 11 mm Hg/cm, P = .017; 994 +/- 185 versus 426 +/- 76 g/cm2, P = .002, respectively). The pattern of left ventricular diastolic filling changed, with a decrease of early peak flow (231 +/- 20 versus 217 +/- 21 mL/s, P = .022), of deceleration time (163 +/- 28 versus 116 +/- 26 seconds, P = .001), and of flow area during rapid filling (105 +/- 15 versus 75 +/- 12 mL, P = .004) and an increase of flow area during atrial contraction (39 +/- 4 versus 88 +/- 9 mL, P = .001). CONCLUSIONS: Our detailed evaluation of left ventricular mechanics demonstrates that cardiomyoplasty has significant multiple beneficial effects on dilated myopathic heart. PMID- 8222172 TI - Influence of left ventricular assist on valvular regurgitation. AB - BACKGROUND: The effects of mechanical left ventricular assist on the nonassisted right ventricle have not been fully elucidated. Current information indicates that the right ventricle benefits from a lower left atrial pressure; however, ventricular septal shifting and increased venous return caused by left ventricular assist impair right ventricular function. Acute intraoperative alterations in mitral and tricuspid valve regurgitation (MR and TR, respectively) may occur as a result of mechanical left ventricular assist but have not yet been documented. METHODS AND RESULTS: Eight patients undergoing implantation of a left ventricular assist device (LVAD) as a bridge to transplantation were studied during surgery by transesophageal echocardiography. MR was present in seven of eight patients, and TR was present in eight of eight patients before LVAD implant (mean MR jet area, 10.6 +/- 2.4 cm2, mean TR jet area, 4.8 +/- 1.0 cm2). Immediately after LVAD placement, MR was still present in seven of eight patients, and TR was present in eight of eight patients (mean MR jet area, 4.2 +/ 0.9 cm2; mean TR jet area, 8.4 +/- 1.9 cm2) (P < .05 preimplant versus postimplant jet area). These changes in MR and TR were associated with a decrease in left ventricular end-systolic dimension (62 +/- 4 versus 48 +/- 3 mm) and an increase in right ventricular end-systolic dimension (31 +/- 4 versus 40 +/- 5 mm) (P < .05 preimplant versus postimplant end-systolic dimension). No patients developed progressive right ventricular failure during 70 to 279 days of LVAD support. CONCLUSIONS: Mechanical left ventricular assist causes an acute decrease in preexisting MR. However, left ventricular assist may acutely worsen TR, presumably by shifting the ventricular septum leftward and increasing venous return to the right ventricle. PMID- 8222173 TI - Significance of gaseous microemboli in the cerebral circulation during cardiopulmonary bypass in dogs. AB - BACKGROUND: Gaseous microemboli during cardiac surgery may damage the brain by reducing cerebral blood flow (CBF). We examined whether the incidence of gaseous microemboli during 150-minute hypothermic (28 degrees C) cardiopulmonary bypass (CPB) adversely affects CBF (radioactive microspheres). METHODS AND RESULTS: Thirty anesthetized dogs were placed on CPB using bubble oxygenators with 50% O2 (n = 10) or 100% O2 (n = 10) to produce a wide range in the number of gaseous microemboli or membrane oxygenators with 50% O2 (n = 10) to avoid microemboli. The number of carotid artery microemboli occurring in a 1-minute interval was counted using a 5-MHz Doppler probe every 15 minutes for the duration of CPB, which lasted 258 +/- 5 minutes. With bubbled 100% O2, the number of microemboli averaged 4.1 +/- 1.7 emboli per minute on normothermic bypass and increased with cooling to 18.3 +/- 4.9 emboli per minute (P < .001). With bubbled 50% O2, the microemboli number was greater on normothermic bypass (19.8 +/- 9.8 emboli per minute, P = .0653 compared with bubbled 100% O2) and increased with cooling (100.3 +/- 18.7 emboli per minute, P < .001) to a greater extent than with bubbled 100% O2 (P < .001). In contrast, with membrane 50% O2, the emboli number was small (0.6 +/- 0.1 emboli per minute) and did not change with CPB temperature. CBF values were not reduced after termination of CPB, even when compared with prebypass values, being 48.3 +/- 7.5 mL/min per 100 g (bubbled 50% O2), 49.6 +/- 4.1 mL/min per 100 g (bubble 100% O2), and 44.5 +/- 2.8 mL/min per 100 g (membrane 50% O2, P = .7581). Similarly, regional perfusion to the cerebellum, hippocampus, and caudal brainstem was not adversely affected by microemboli. After CPB, cortical biopsies demonstrated no difference among groups with respect to lactate (P = .1753), energy charge (P = .5179), and brain water content (P = .939). Retinal histopathology indicated no differences among groups. CONCLUSIONS: These results indicate that: (1) the incidence of gaseous microemboli during hypothermia increases when a bubble oxygenator is used, and (2) global CBF and regional brain perfusion are not adversely affected by numerous gaseous microemboli. PMID- 8222174 TI - Reoperative coronary artery bypass surgery. Improved preservation of myocardial function with retrograde cardioplegia. AB - BACKGROUND: Cardiac-related mortality remains high for reoperative coronary artery bypass graft surgery (rCABG) compared with first-time surgery (fCABG). Retrograde cardioplegia (RC) has been suggested but not proven to improve the results for rCABG. METHODS AND RESULTS: We therefore reviewed the records of 240 consecutive patients who had undergone rCABG at our institution since 1988. The interval to reoperation was 9.1 +/- 4.2 years (mean +/- SD), with a range from 0.2 to 18 years. Only 46% of grafts were patent at the time of rCABG. The incision to cardiopulmonary bypass (CPB), incision to cross-clamp (XCL), and XCL per graft time intervals were significantly prolonged compared with 100 consecutive fCABG patients operated on during the same time period. Blood utilization was also significantly increased in rCABG compared with fCABG patients. Overall operative mortality was 5.8% and 0% for rCABG and fCABG patients, respectively (P < .05). High-risk criteria (emergency surgery, angina at rest requiring intravenous nitroglycerin or intra-aortic balloon pump [IABP] support [urgent surgery], recent [<21 days] myocardial infarction, or ejection fraction < 30%) were noted in 136 rCABG patients (57%) and 28 fCABG patients (28%) (P < .001). Profound postoperative myocardial dysfunction (postoperative IABP dependence) occurred in only one of 104 low-risk patients (1%), compared with 14 of 136 high-risk patients (10%) (P < .005). Operative mortality was noted in 13 high-risk patients (9.5%) compared with one low-risk patient (1%) (P < .005). RC was used in 80 patients without complication. Postoperative IABP dependence developed in only 2 of 53 high-risk/RC patients (3.8%) compared with 12 of 83 high-risk/non-RC patients (14.5%) (P < .05). At follow-up, rCABG and fCABG patients enjoyed similar symptomatic improvement. CONCLUSIONS: We conclude that retrograde cardioplegia, possibly by minimizing the increased ischemia associated with rCABG, improves the results of rCABG, specifically in regard to preventing profound myocardial dysfunction in high-risk patients. PMID- 8222175 TI - Protection of the stunned myocardium. Selective nucleoside transport blocker administered after 20 minutes of ischemia augments recovery of ventricular function. AB - BACKGROUND: Metabolic interventions capable of preventing ventricular dysfunction "stunning" or accelerating its functional recovery have potential clinical importance. Myocardial protection of the stunned myocardium has not been documented when drugs were administered only during postischemic reperfusion. The role of ATP depletion and release of purines in myocardial injury was assessed using the selective nucleoside transport blocker p-nitrobenzylthioinosine (NBMPR) in a combination with specific adenosine deaminase inhibitor erythro-9-[hydroxy-3 nonyl]adenine (EHNA) administered during reperfusion after reversible ischemic injury. METHODS AND RESULTS: Sixteen anesthetized dogs were instrumented with minor axis sonocrystals and intraventricular Millar. Ventricular performance was determined, off bypass, from the slope of the relationship between stroke-work and end-diastolic length as a sensitive and load-independent index of contractility within physiological range. Hearts were subjected to 20 minutes' warm global ischemia and reperfused with warm blood treated with either saline (control group, n = 8) or saline containing 100 mumol/L EHNA and 25 mumol/L NBMPR (EHNA/NBMPR-treated group, n = 8). Myocardial biopsies were collected and analyzed for ATP and metabolites using high-performance liquid chromatography. Warm ischemia induced significant depletion of ATP (P < .05 versus preischemia) and accumulation of inosine at the end of ischemia (> 90% of total nucleosides) in both groups. Complete functional recovery was observed in the EHNA/NBMPR treated group (P < .05 versus control group). CONCLUSIONS: Selective entrapment of adenine nucleosides during postischemic reperfusion attenuated ventricular dysfunction (stunning) after brief global ischemia. It is concluded that nucleoside transport plays an important role in myocardial stunning, and its blockade augmented myocardial protection against reperfusion injury. Selective entrapment of endogenous inosine, generated during ischemia, represents an attractive therapeutic approach to the alleviation of postischemic dysfunction mediated by reperfusion in a wide spectrum of ischemic syndromes, including percutaneous transluminal coronary angioplasty and coronary artery bypass graft surgery. PMID- 8222176 TI - Comparison of cold versus warm cardioplegia. Crystalloid antegrade or retrograde blood? AB - BACKGROUND: To evaluate the efficacy of warm versus cold and antegrade versus retrograde cardioplegia, 163 patients were randomized in sequence in three groups and underwent isolated coronary artery bypasses (mean, 4 grafts/patients) alternating in sequence. METHODS AND RESULTS: The demographic profiles were identical. Cold crystalloid cardioplegia (group 1) was delivered through the aortic root intermittently. The warm retrograde (group 2) was started antegrade with high potassium solution until the heart stopped. It was continued with retrograde coronary sinus perfusion, 100 mL/min of warm blood. Cold retrograde blood cardioplegia (4:1) (group 3) was started antegrade and continued retrograde through the coronary sinus. The heart temperature was kept at 10 degrees to 15 degrees C. Patients were evaluated intraoperatively and postoperatively for (1) supraventricular tachycardia, (2) ECG changes, (3) lactate dehydrogenase and total CPK and isoenzyme (MB) studies, and (4) hemodynamic studies in the intensive care unit. Warm retrograde and cold retrograde patients had sampling of the ascending aorta (antegrade) and the coronary sinus (retrograde) measuring pH, A-VO2 differences, and CK enzyme leak. RESULTS: The incidence of supraventricular tachycardia was 29% in group 2, 22% in group 1, 18% in group 3; not significant (NS). CPK isoenzyme MB fraction showed identical levels (NS). The warm heart consumed 1.3 to 1.6 mL O2/100 mL flow, while the cold group 3 showed 0.5 to 0.6 mL O2/100 mL flow (P < .001). Cold crystalloid cardioplegia (group 1) was similar to group 3 (0.3 mL O2/100 mL). All three groups were similar hemodynamically (cardiac output, cardiac index, left ventricular stroke work index). Two of 163 patients died in group 2. Four sustained stroke, three in group 2, one with a fatal outcome. CONCLUSIONS: Continuous warm cardioplegia (group 2) did not provide better myocardial protection despite that no CK-MB isoenzyme leak was demonstrated intraoperatively. Intermittent cold crystalloid cardioplegia and cold retrograde provided a clearer operative field and motionless heart. As long as O2 was adequately supplied, under 90 minutes' cross-clamp time, cold crystalloid cardioplegia and cold retrograde blood cardioplegia is safe under hypothermic conditions, whereas warm cardioplegia requires continuous uninterrupted technique with oxygen delivery. PMID- 8222177 TI - Transposition of chordae in mitral valve repair. Mid-term results. AB - BACKGROUND: Prolapse of the anterior mitral leaflet, unlike a posterior prolapse, is a difficult lesion to repair. Leaflet plication and triangular resection are satisfactory techniques only in case of a limited prolapse. Chordal replacement has also been proposed but uses foreign material. The purpose of this report is to assess the results of transposition of chordae for the correction of mitral regurgitation (MR) caused by anterior leaflet prolapse. METHODS AND RESULTS: Between January 1986 and December 1990, 44 adult patients with MR caused by anterior leaflet prolapse underwent repair with transposition of chordae as one of the techniques. This population was retrospectively studied to assess the early and late results of this procedure. Chordae were transferred from the posterior to the anterior leaflet (n = 25) or from an intermediary to a free edge position on the anterior leaflet (n = 21) (two patients underwent both procedures). Two patients died (4.5%). None required early reoperation. Follow-up was complete and ranged from 18 to 82 months (mean, 40.2 +/- 19 months). No patient died during follow-up. Two patients were reoperated on 6 and 8 months after surgery for recurrent MR unrelated to chordal transfer disruption. Doppler echocardiographic studies were available in 95% of the cases at latest follow-up and showed no or minimal MR (0 to 1/4) in 87.5% of the patients and mild MR (2/4) in 12.5%. CONCLUSIONS: Transposition of chordae appeared to be a simple and safe procedure for correction of anterior mitral prolapse. Transposition of chordae allowed extension of the indications of valve repair. A longer follow-up will be necessary to draw firm conclusions, but mid-term results are encouraging. PMID- 8222178 TI - Acadesine improves surgical myocardial protection with blood cardioplegia in ischemically injured canine hearts. AB - BACKGROUND: Adenosine is a cardioprotective autacoid that exerts receptor mediated protection from ischemia/reperfusion injury. In ischemically injured hearts, avoidance of ischemia/reperfusion injury with hypothermic chemical cardioplegia may be incomplete, and consequently, postischemic left ventricular (LV) function may be severely depressed and chamber stiffness increased. This study tested the hypothesis that the adenosine-regulating agent acadesine improves myocardial protection with hypothermic blood cardioplegia (BCP), resulting in better postischemic LV function and diastolic characteristics in hearts injured by 45 minutes of normothermic global ischemia. METHODS AND RESULTS: Eighteen anesthetized (350 micrograms fentanyl citrate, 5 mg diazepam) dogs on total vented bypass were randomized to receive vehicle (n = 5), low-dose acadesine (LDA, 0.125 mg.kg-1.min-1, n = 6) or high-dose acadesine (HDA, 0.5 mg.kg-1.min-1, n = 7) continuously infused 30 minutes before global ischemia and discontinued 10 minutes after aortic cross-clamp removal. Hearts were protected with cold (4 degrees C) multidose (every 20 minutes) potassium BCP, which contained saline vehicle, 1 mg/L acadesine (LDA), or 4 mg/L acadesine (HDA) for a total of 1 hour of cardioplegic arrest. Postischemic LV function, assessed by the slope of the end-systolic pressure-volume (impedance catheter) relation, was depressed by 34 +/- 7% of baseline (5.6 +/- 1.0 versus 2.7 +/- 0.7 mm Hg/mL, P < .05) in vehicle. With LDA, there was variable improvement in postischemic function (5.1 +/- 1.3 versus 3.6 +/- 0.6 mm Hg/mL, P = .26 versus baseline). In contrast, there was complete postischemic functional recovery with HDA (5.9 +/- 0.6 versus 5.2 +/- 0.8 mm Hg/mL, P = .54). Postischemic chamber stiffness was preserved in both LDA and HDA. CONCLUSIONS: We conclude that the higher dose of acadesine improves myocardial protection when used as a pretreatment and BCP adjuvant, resulting in better postischemic LV systolic function and diastolic characteristics. PMID- 8222179 TI - The effects of warm versus cold blood cardioplegia on endothelial function, myocardial function, and energetics. AB - BACKGROUND: Recent interest in the use of normothermic blood cardioplegia is based on theoretical advantages over the traditional method of hypothermic myocardial protection. This study was designed to compare the effects of warm and cold blood cardioplegia on left ventricular functions and energetics and coronary responsiveness. MATERIALS AND METHODS: Two groups of mongrel dogs (n = 7 each) underwent either normothermic cardiopulmonary bypass (CPB) with continuous warm (37 degrees C) blood cardioplegia or hypothermic (26 degrees C) CPB with a single dose of cold (4 degrees C) blood cardioplegia supplemented with topical cooling during 30 minutes of aortic clamping. There was no deterioration in the endothelium-dependent and -independent coronary relaxation as tested by the infusion of acetylcholine and nitroglycerin after cardioplegic arrest for either group. At 60 minutes of reperfusion, both groups had complete recovery of left ventricular contractility as measured by the preload recruitable stroke work area derived from the measurement of the ventricular pressure (micromanometer catheter) and volume (conductance catheter) relation. The analysis of myocardial energetics in terms of the myocardial oxygen consumption-pressure volume area relation did not reveal any significant changes between the y-intercepts and the slopes of the two groups. CONCLUSIONS: For 30 minutes of aortic cross-clamp time, continuous warm cardioplegia did not provide any benefit over a single injection of cold cardioplegia in coronary endothelial and smooth muscle function, myocardial function, and energetics. PMID- 8222180 TI - Nitroglycerin improves functional recovery of neonatal lamb hearts after 2 hours of cold ischemia. AB - BACKGROUND: Vascular events during initial reperfusion after a period of hypothermic cardioplegic ischemia may play an important role in the recovery of mechanical function. Prior studies have shown (1) that dysfunction of the postischemic myocardium after normothermic ischemia can be ameliorated by vasodilation and (2) that endothelial dysfunction in the coronary bed and mechanical dysfunction were correlated after hypothermic ischemia. The present study was undertaken to further explore the relation between vascular events during reperfusion and the recovery of mechanical function after hypothermic ischemia by the infusion of the endothelium-independent vasodilator nitroglycerin during reperfusion. METHODS AND RESULTS: We assessed the effect of an infusion of nitroglycerin on the recovery of isolated blood-perfused neonatal lamb hearts arrested for 2 hours with 15 degrees C K+ cardioplegic solution, comparing 8 hearts receiving a nitroglycerin infusion with 9 control hearts. After 30 minutes of reperfusion, nitroglycerin-infused hearts compared with control hearts had better percent recovery of maximum left ventricular developed pressure (83.5 +/- 8.1% vs 73.6 +/- 8.9%, mean +/- SD), left ventricular dP/dt (77.2 +/- 7.7% vs 67.4 +/- 10.3%), coronary blood flow (141.2 +/- 30.6% vs 84.4 +/- 10.4%), and myocardial oxygen consumption (100.7 +/- 51.3% vs 71.2 +/- 18.6%), (all except myocardial oxygen consumption with P < .05). In each heart, we also tested the coronary vascular resistance response to 10(-6) mol/L acetylcholine infusion to assess endothelial function. The recovery of the coronary vascular resistance response to acetylcholine was reduced after reperfusion in both groups compared with the preischemic response to acetylcholine (nitroglycerin, 14.9 +/- 53.8%; control, 13.4 +/- 38.3%). CONCLUSIONS: These results suggest that the effects of hypothermia and reperfusion on recovery of mechanical function after hypothermic ischemia can be offset by an infusion of nitroglycerin. Vasospasm secondary to endothelial dysfunction may play an important role in cardiac recovery after hypothermic ischemia. PMID- 8222181 TI - Inhibition of neutrophil adherence improves postischemic ventricular performance of the neonatal heart. AB - BACKGROUND: Reduction of the leukocyte population during postischemic coronary reperfusion results in decreased neutrophil-mediated tissue injury. However, the importance of leukocyte adhesion to coronary endothelium for postischemic ventricular dysfunction after global hypothermic myocardial ischemia is unknown. Neutrophil integrins (CD11b/CD18) upregulate in response to cardiopulmonary bypass and ischemic stress, and their role in generating postoperative ventricular dysfunction was examined in this study. METHODS AND RESULTS: An in vivo, in situ model of neonatal cardiac surgery was established in which neutrophil adherence was manipulated by administering NPC 15669 (an inhibitor of neutrophil CD11b/CD18 surface receptor upregulation). Seventeen 3- to 5-day-old piglets (8 controls and 9 NPC 15669-treated animals) were instrumented with a coronary sinus catheter, sonomicrometry crystals across the short axis of the left ventricle (LV), and a micromanometer positioned in the LV. Hearts were subjected to 90 minutes of hypothermic ischemia after a single dose of cold crystalloid cardioplegia. Myocardial granulocyte accumulation during ischemia and reperfusion was reduced in NPC animals compared with controls (myeloperoxidase activity, 43.4 +/- 2.6 and 75.8 +/- 6.3 mumol/10 mg tissue, respectively; P < or = .005). This was associated with a reduction in coronary vascular resistance in NPC animals compared with controls (P < or = .02) and decreased release of myocardial creatine phosphokinase throughout reperfusion (P < or = .05). NPC animals demonstrated an improved preservation of the end-systolic pressure-volume relation after discontinuation of cardiopulmonary bypass (71 +/- 6% and 96 +/- 6% at 60 minutes, respectively; P < or = .05). There was no difference in ventricular compliance between groups. CONCLUSIONS: These data demonstrate that inhibition of neutrophil CD11b/CD18 surface adherence receptor upregulation reduces granulocyte accumulation in myocardium after hypothermic global ischemia, reduces myocyte damage, and improves ventricular systolic function. PMID- 8222182 TI - Effects of adenosine infusion during reperfusion after cold cardioplegic ischemia in neonatal lambs. AB - BACKGROUND: Postischemic infusion of adenosine (Ado) has been postulated to reduce reperfusion injury after normothermic ischemia, but the effect of Ado after hypothermic ischemia is unknown. METHODS AND RESULTS: We infused Ado (350 mumol/L), Ado + Ado antagonist theophylline (Ado + T, 7 mmol/L), A1 agonist cyclohexyl-Ado (0.25 mumol/L), A2 agonist cyclopropyl carboxamide Ado (0.25 mumol/L), or blood alone (group C) during the first 20 minutes of reperfusion after 2 hours of cold cardioplegic ischemia using an isolated, blood-perfused neonatal lamb heart model (n = 8 in each group). At 30 minutes of reperfusion, left ventricular (LV) maximum developed pressure (DP), dP/dt, -dP/dt, maximum DP at V10 (volume that gives LVEDP of 10 mm Hg at baseline measurement), dP/dt at V10, and LV stiffness constant (KA) at V10 were measured. Coronary blood flow and oxygen consumption (MVO2) were also measured to evaluate the metabolic recovery. Groups Ado, A1, and A2 showed better functional recovery than group C (DP for group C, 74.6 +/- 5.60%; group Ado, 94.2 +/- 10.7%; group A1, 89.3 +/- 4.40%; group A2, 86.7 +/- 9.00%; P < .01), but theophylline offset the Ado effect (Ado + T, 74.6 +/- 6.4%). Coronary blood flow was higher in groups Ado + T and A2 than group C (group C, 156.7 +/- 45.7%; group Ado + T, 238.0 +/- 41.0%; group A2, 258.6 +/- 70.3%; P < .05). MVO2 was higher in groups Ado, Ado + T, A1, and A2 than group C (group C, 57.7 +/- 11.0%; group Ado, 91.5 +/- 20.1%; group Ado + T, 210.7 +/- 60.0%; group A1, 82.8 +/- 31.9%; group A2, 83.5 +/- 12.8%; P < .05). CONCLUSIONS: Ado infusion improved recovery of mechanical function after hypothermic ischemia, and its beneficial effect appears to involve both A1 and A2 receptors. Since group Ado + T had high coronary blood flow but not improved systolic function and group A1 had low coronary blood flow and improved systolic function, coronary vasodilation alone is not responsible for this beneficial effect. Ado may be a useful agent after hypothermic ischemia in the immature heart, although the precise mechanisms remain unclear. PMID- 8222183 TI - Interleukin-1 alpha preconditioning reduces myocardial ischemia reperfusion injury. AB - BACKGROUND: Interleukin-1 (IL-1) has been shown to induce superoxide dismutase (SOD) activity and to express heat shock protein (HSP). Since the reperfusion of ischemic heart is associated with the reduction of antioxidative enzymes including SOD and expression of HSP, it was hypothesized that IL-1 could be beneficial against ischemic reperfusion injury. METHODS AND RESULTS: Rats were injected with recombinant IL-1 alpha (30 micrograms/kg IP); after 48 hours, they were anesthetized and hearts were removed, isolated, and perfused by the Langendorff technique. Myocardial functions were studied by measuring left ventricular developed pressure (LVDP) and its maximum first derivative (LV dP/dt), and cellular injury was studied by estimating creatine kinase (CK) release. Induction of the expression of HSP27 mRNA and HSP27 protein was examined by Western blot analysis and Northern blot analysis, respectively. Antioxidant enzymes were assayed by enzymatic analysis. Our results indicated reduction of ischemic reperfusion injury by IL-1 alpha, as evidenced by better recovery in postischemic ventricular functions (LVDP [mm Hg]: control, 63 +/- 14; IL-1, 102 +/- 11; P < .05), increased coronary flow (mL/min) (control, 2.93 +/- 0.58; IL-1, 5.17 +/- 0.43; P < .03), and reduced creatine kinase release (IU/L) (control, 110 +/- 5.78; IL-1, 81.76 +/- 7.71; P < .01). IL-1 alpha induced the expression of HSP27 mRNA within 2 hours as examined by Northern blot analysis and the expression of HSP27 after 48 hours. In addition, hearts pretreated with IL-1 alpha for 48 hours and then subjected to 30-minute ischemia and 60-minute reperfusion enhanced the activities (nmol/min/mg protein) of Cu/Zn SOD (control, 1.55 +/- 0.22; IL-1 alpha, 2.92 +/- 0.04; P < .004), Mn-SOD (control, 4.54 +/- 0.19; IL-1 alpha, 6.33 +/- 0.09, P < .001), catalase (control, 15.53 +/- 0.37; IL 1 alpha, 21.67 +/- 0.72; P < .002), glutathione peroxidase (control, 17.49 +/- 0.35; IL-1 alpha, 25.87 +/- 0.58; P < .001), and glucose-6-phosphate dehydrogenase (control, 22.71 +/- 0.44; IL-1 alpha, 29.53 +/- 0.48; P < .001). CONCLUSIONS: The results of this study indicate that low doses of IL-1 alpha can be used as a therapeutic agent to precondition a heart from ischemia reperfusion injury. PMID- 8222184 TI - Do patients with less than 'echo-perfect' results from mitral valve repair by intraoperative echocardiography have a different outcome? AB - BACKGROUND: Not all valve repairs for mitral regurgitation (MR) have a perfect result, with no MR on postpump intraoperative echocardiography. Although more than 2+ MR by postpump echocardiography has led to second pump runs for further surgery in 6% to 8% of our patients, those left with 1+ or 2+ MR, traditionally an acceptable result, have not previously been evaluated for clinical outcome. METHODS AND RESULTS: Among 530 patients undergoing mitral repair during 1987 to 1989, 76 patients with 1+ or 2+ MR by postpump intraoperative echocardiography were compared with 76 patients who had equivalent age, sex, left ventricular function, and concomitant surgery but who had no MR by postpump echo. In-hospital morbidity measured by the frequency of respiratory complications, strokes, time in intensive care unit, and duration of hospital stay was actually higher in the patients with no MR after repair. Hospital mortality was not significantly different. There were no significant differences in posthospital mortality (4 year survival estimate of 86% in both groups), thromboembolic events, hospitalizations for heart failure, or functional class, although more patients in the group with no MR after repair were using diuretics. There was a trend toward more reoperations in patients with 1+ or 2+ MR compared with those with no MR by intraoperative echocardiography after repair (4-year estimates of freedom from reoperation, 83% versus 94%; adjusted risk ratio, 3.30). CONCLUSIONS: The presence of 1+ or 2+ MR by postpump intraoperative echocardiography does not confer increased morbidity or mortality. However, the trend toward more reoperations suggests the need for close follow-up for possible recurrence of MR. These data support our low threshold for performing further surgery during second pump runs. PMID- 8222185 TI - Coronary endothelial injury after cardiopulmonary bypass and ischemic cardioplegia is mediated by oxygen-derived free radicals. AB - BACKGROUND: Cardiopulmonary bypass and crystalloid cardioplegia may lead to endothelial dysfunction in the coronary microcirculation. The aim of the present study was to examine whether the alteration of endothelium-dependent microvascular responses may be related to the generation of oxygen-derived free radicals. METHODS AND RESULTS: Pigs (30 kg) were heparinized and placed on cardiopulmonary bypass. The hearts were arrested for 1 hour with either plain hypothermic, hyperkalemic (25 mEq/L) crystalloid cardioplegic solution (n = 10) or crystalloid cardioplegic solution containing either deferoxamine (n = 8) or manganese superoxide dismutase (n = 6). Hearts were then reperfused for 1 hour while the pigs were separated from cardiopulmonary bypass. Noninstrumented pigs were used as controls (n = 8). Coronary microarteries (120 to 190 microns in diameter) were studied in vitro in a pressurized (40 mm Hg), no-flow state with videomicroscopy and electronic dimension analysis. After precontraction of microvessels, the endothelium-dependent and -independent agents were applied extraluminally. Serotonin caused a slight dilation of control vessels (percent dilation of acetylcholine-induced preconstriction at 10 mumol/L drug concentration, 5 +/- 8%; P < .05 versus crystalloid cardioplegia group) and a significant contractile response after crystalloid cardioplegia (-28 +/- 10%). Bradykinin elicited near complete relaxation of control vessels (96 +/- 3%, P < .05), whereas it caused considerably less relaxation after cardioplegia (33 +/- 9%). The addition of either deferoxamine or superoxide dismutase to the cardioplegic solution significantly (but not completely) preserved vasomotor responses of coronary microvessels to serotonin (9 +/- 6% and 11 +/- 4%, respectively; P < .05) or bradykinin (72 +/- 4% and 87 +/- 3%, respectively; P < .05). Endothelium-independent relaxations of vessels in response to sodium nitroprusside were similar in all groups. CONCLUSIONS: Either the hydroxyl radical synthesis inhibitor deferoxamine or manganese superoxide dismutase preserves endothelium-dependent relaxation during crystalloid cardioplegia reperfusion. Therefore, ischemic cardioplegia-reperfusion-induced endothelial dysfunction is at least partially mediated by the generation of oxygen-derived free radicals. PMID- 8222186 TI - Induction of interleukin-8 expression during cardiopulmonary bypass. AB - BACKGROUND: Patients undergoing cardiopulmonary bypass (CPB) are known to suffer from a postsurgical systemic inflammatory response, the nature of which remains to be fully elucidated. Interleukin-8 (IL-8) is a newly described, powerful leukocyte chemotactic factor known to be generated after stimulation of interleukin-1 (IL-1). As we have previously documented the generation of IL-1 beta after CPB, it followed that IL-8 generation should be measured in a comparable group of patients. METHODS AND RESULTS: Twenty-two adult patients aged 41 to 81 years undergoing coronary revascularization were studied for measurements of C3a, C5a, IL-1, IL-8, and OH(.). Blood was collected before surgery, after CPB, and at 24, 48, and 72 hours. A significant increase in IL-1 beta and IL-8 was detected in circulating leukocytes with peak levels at 24 hours after bypass. No IL-1 beta or IL-8 antigen was detected at any time in patient plasma. Comparable to interleukin generation, human complement-derived C5a also peaked after 24 hours, whereas C3a was increased dramatically immediately after CPB, followed by a decline at 24 hours and a progressive increase over the next 48 hours. CONCLUSIONS: The results demonstrated for the first time the presence of cell-associated IL-8 in CPB patients. This suggests that this powerful polymorphonuclear and T-lymphocyte chemotactic factor may be an important element in leukocyte activation and recruitment after CPB. PMID- 8222187 TI - Heparin treatment enhances the recovery of neoendothelial acetylcholine-induced vascular relaxation after balloon catheter injury in the rabbit aorta. AB - BACKGROUND: After catheter injury, the neoendothelium that grows is abnormal in morphology and in acetylcholine-induced generation of endothelium-derived relaxing factor (EDRF). Heparin has been shown to have stimulatory effects on vascular endothelial growth in vitro. Its effect in vivo on neoendothelial cell morphology and metabolism after injury has not been described. We investigated the effect of heparin treatment on the neoendothelium formed after injury. METHODS AND RESULTS: Four groups of New Zealand White rabbits were studied. Group 1 rabbits underwent catheter denudation and were killed 4 weeks after injury without receiving treatment (NO Tx, n = 8). Groups 2 and 3 underwent similar aortic injury, received 2 weeks of treatment with either heparin (n = 7) or low molecular weight heparin (LMWH, n = 5), and were killed at 4 weeks. Group 4 underwent sham operation (SHAM, n = 8). EDRF generation was determined by the relaxation of precontracted aortic rings in an organ bath in response to acetylcholine. The heparin-treated group exhibited a significant improvement in acetylcholine-induced relaxation (27%) versus both LMWH-treated (14%, P = .035) and untreated groups (11%, P = .004), although relaxation was only 50% of that observed in the uninjured control vessels (52%, P = .001). The neoendothelium formed in the heparin-treated group exhibited a more normal histological appearance and was aligned with the direction of blood flow as compared with that observed in the untreated or LMWH-treated groups. CONCLUSIONS: These results demonstrate that in vivo heparin administration enhanced the recovery of EDRF generation and augmented normalization of the morphologic appearance of the neoendothelium. PMID- 8222188 TI - Heparin binding proteins. Contribution to heparin rebound after cardiopulmonary bypass. AB - BACKGROUND: Heparin rebound, the reappearance of anticoagulant activity after adequate neutralization with protamine, can lead to excessive postoperative bleeding after cardiac surgery. We investigated the mechanism of heparin rebound by using chemically modified heparin that lacks anticoagulant activity (low affinity heparin) but that is able to displace protein-bound anticoagulantly active heparin. METHODS AND RESULTS: Sixteen patients undergoing elective cardiac surgery were given heparin (400 U/kg) to achieve an activated clotting time (ACT) > 400 seconds. After cardiopulmonary bypass, protamine sulfate was given (by heparin-ACT dose-response curve) to return the ACT to prebypass times (preoperative, 160 +/- 9 seconds; postoperative, 156 +/- 17 seconds). Blood samples were obtained serially for 24 hours and assayed for thrombin clotting time (TCT) and heparin activity using an anti-factor Xa assay. The TCT and anti factor Xa activity were consistently and abnormally elevated for the first 6 hours after surgery. The anti-factor Xa activity increased fourfold after the addition of low-affinity heparin (essentially free of anti-factor Xa activity), indicating that anticoagulantly active heparin persisted in the circulation after protamine neutralization bound nonspecifically to plasma proteins. Blood loss correlated with postoperative TCTs. CONCLUSIONS: Our findings demonstrate that heparin anticoagulant activity persists for up to 6 hours after surgery despite apparent protamine neutralization. The observation of the marked increase in plasma anti-factor Xa activity after the addition of low-affinity heparin suggests that after its administration, a large proportion of the heparin binds to plasma proteins and is incompletely removed by protamine. After protamine is cleared, the protein-bound heparin dissociates slowly and binds to anti-thrombin III to produce an anticoagulant effect. PMID- 8222189 TI - A comparative study of heparin responses in arterial and venous thromboembolism using molecular markers for thrombosis. AB - BACKGROUND: Compared with the therapy of venous thromboembolism, there is little consensus regarding guidelines for heparin anticoagulation of patients with arterial thrombosis. This study aimed to identify the quantitative differences in the activation of the coagulation cascade and platelets in these two syndromes and to characterize their specific biological responses to heparin therapy. METHODS AND RESULTS: Eighteen patients receiving intravenous heparin to treat venous (n = 9) or arterial (n = 9) thromboembolism were prospectively studied for an average of 4 days each. Clinical responses to treatment, activated partial thromboplastin time (aPTT), and molecular markers for thrombosis were measured regularly. Although both groups received equivalent doses of heparin (approximately 1100 units/h), the resulting aPTTs and plasma heparin activity were significantly lower in the arterial patients (P < .05 and P < .01, respectively). The plasma levels of beta-thromboglobulin (a marker for platelet activation and granule release) were significantly higher in the arterial patients (109 +/- 9.5 versus 79 +/- 7.1 ng/mL, mean +/- SEM, P < .05). In vivo fibrin formation, as evidenced by plasma levels of fibrinopeptide A, was less effectively suppressed in the patients with arterial versus venous thrombosis (18.5 +/- 3.2 versus 10.4 +/- 2 ng/mL, P < .05). Prothrombin fragments 1 + 2, a marker for prothrombinase complex activity, was nearly normal in both heparinized groups. CONCLUSIONS: The anticoagulant response to heparin is blunted in patients with arterial thrombosis, at least in part by the antagonistic actions of increased platelet activation. Comparing arterial with venous thrombosis, higher doses of heparin on the average may be required to achieve comparable aPTTs, plasma heparin activity, and comparable suppression of fibrin formation. PMID- 8222190 TI - Nafamostat mesilate reduces blood loss during open heart surgery. AB - BACKGROUND: Nafamostat mesilate (FUT-175) is a protease inhibitor inactivating coagulation, fibrinolysis, and platelet aggregation. A prospective, randomized trial was performed to assess the efficacy of FUT-175 in the reduction of postoperative bleeding tendency. METHODS AND RESULTS: FUT-175 was infused at a rate of 40 mg/h during cardiopulmonary bypass (CPB) along with 300 IU/kg of heparin in 20 patients undergoing aortocoronary bypass surgery (FUT group). This group was compared with another group of 20 patients undergoing aortocoronary bypass surgery, who were given only heparin (control group). Blood concentrations of FUT-175 and activated clotting time increased after cooling, peaking at 2050 +/- 1190 ng/mL and 2136 +/- 983 seconds at the lowest temperature and recovered after rewarming to values of 166 +/- 118 ng/mL and 510 +/- 148 seconds, respectively, at the end of CPB. In the FUT group, platelet counts were significantly higher at the end of CPB than those in the control group (168 +/- 10 versus 136 +/- 9 x 10(3)/mm3, P < .05). In the FUT group, serum concentrations of beta-thromboglobulin (307 +/- 102 versus 537 +/- 391 ng/mL, P < .05), PIC (3.6 +/- 1.7 versus 5.8 +/- 3.8 micrograms/mL, P < .05) and FDP (20.2 +/- 7.0 versus 41.4 +/- 11.9 ng/mL, P < .01) were significantly lower than those in the control group at the end of CPB. However, serum concentrations of TAT, FPA, and FPB beta 15-42 showed no significant differences between groups. FUT-175 significantly reduced heparin dosage and 24-hour postoperative blood loss (19,200 +/- 3200 versus 29,100 +/- 7700 IU, P < .01, and 382 +/- 129 versus 576 +/- 375 mL, P < .05). CONCLUSIONS: FUT-175 inhibits fibrinolysis and preserves platelet counts and function during CPB and reduces blood loss during open heart surgery. PMID- 8222191 TI - Cost-effectiveness of radiofrequency ablation compared with other strategies in Wolff-Parkinson-White syndrome. AB - BACKGROUND: Patients with Wolff-Parkinson-White syndrome fall into four risk groups: those with (1) prior cardiac arrest, (2) paroxysmal supraventricular tachycardia or atrial fibrillation (PSVT/AF) with hemodynamic compromise, (3) PSVT/AF without hemodynamic compromise, and (4) no symptoms. METHODS AND RESULTS: For each group, we developed a cost-effectiveness analysis examining five clinical management strategies: (1) observation, (2) observation until a cardiac arrest dictates the need for therapy, (3) initial drug therapy guided by noninvasive monitoring, (4) initial radiofrequency ablation (RFA), and (5) initial surgical ablation. We used a Markov simulation model to estimate life expectancy and costs for patients whose ages are between 20 and 60 years. The model includes the risks of cardiac arrest, PSVT/AF, drug side effects, procedure related complications and mortality, the efficacy of drugs and RFA, and costs. Based on literature and expert opinion, we assumed that the annual risks of cardiac arrest are 0.01%, 0.05%, and 0.5%, respectively, in patients who are asymptomatic, who had PSVT/AF without hemodynamic compromise, or who had PSVT/AF with hemodynamic compromise. We also assumed that RFA has an overall efficacy of 92% in preventing cardiac arrest and arrhythmias. Our model predicts that RFA should yield a life expectancy greater than or equal to other strategies. In cardiac arrest survivors and patients who have had PSVT/AF with hemodynamic compromise, our model suggests that RFA should both prolong survival and save resources. For patients with PSVT/AF without hemodynamic compromise, the marginal cost-effectiveness of attempted RFA (followed by conservative treatment if the RFA fails) ranges from $6600 per quality-adjusted life year (QALY) gained for 20 year-old patients to $19,000 per QALY gained for 60-year-old patients. For asymptomatic patients, RFA costs from $174,000 per QALY gained for 20-year-old patients to $540,000 per QALY gained for 60-year-old patients. CONCLUSIONS: Our analysis supports the emerging practice of performing RFA in patients with Wolff Parkinson-White syndrome who survive cardiac arrest or who experience PSVT/AF but also supports the current practice of observing asymptomatic patients. PMID- 8222192 TI - Thoracoscopic placement of implantable cardioverter-defibrillator patch leads in sheep. AB - BACKGROUND: A thoracoscopic technique was developed for the placement of commercially available implantable cardioverter-defibrillator (ICD) patch leads in sheep. METHODS AND RESULTS: Small ICD patch leads (13.5 cm2, A-67) were placed thoracoscopically in sheep (n = 5) that had survived coronary artery ligation from a previous experiment. The technique used three small incisions in the left chest. After lysis of adhesions, the ICD patch lead was introduced through a mediastinoscope. The ICD patch lead was secured in the extrapericardial position with surgical clips placed in the four corners of the ICD patch lead. After 2 weeks, a median sternotomy was performed, and ICD patch leads were reexamined for positioning. Extensive fibrosis was noted to adhere the ICD patch lead to the pericardium. The surgical clips were found intact in all animals without noticeable migration of patch lead position. There was no mortality related to ICD patch lead placement, and estimated blood loss was less than 30 mL without use of cautery. CONCLUSIONS: Commercially available ICD patch leads may be reliably and safely placed with minimal patch migration in sheep using thoracoscopic techniques. PMID- 8222193 TI - Isolated lung transplantation for patients with Eisenmenger's syndrome. AB - BACKGROUND: Eisenmenger's syndrome remains one of the greatest challenges in lung transplantation. METHODS AND RESULTS: Since October 1990, seven such patients with Eisenmenger's syndrome received isolated pulmonary grafts (six double lungs and one single lung). Mean patient age was 32 +/- 6 years (two men and five women). The preoperative mean pulmonary arterial pressure was 90.7 +/- 31.2 mm Hg, and the ventriculoscintigram showed markedly enlarged right ventricle and normal left ventricular function with ejection fraction of 0.660 +/- 0.115. Three atrial septal defects and four patent ducti arteriosus were repaired concomitantly. Excised lung histology showed plexogenic pulmonary arteriopathy with Heath-Edwards' grade 4 through 6. One double lung patient who had preexisting systemic vascular collapse died intraoperatively. The other six patients tolerated transplantation, and on the first operative day, mean pulmonary artery pressure decreased to 22.4 +/- 7.3 mm Hg (P < .002) and gas exchange was acceptable with an arterial/alveolar oxygen tension ratio of 0.47 +/ 0.15. Two patients died of mediastinal and pulmonary infection. The follow-up for the four survivors ranged from 13 to 25 months after transplantation. CONCLUSIONS: Our preliminary experience shows that concomitant isolated lung transplantation with cardiac repair could be a viable therapeutic option for patients with Eisenmenger's syndrome and normal left ventricular function. Dynamic right ventricular outflow obstruction is a potential hemodynamic problem in these pulmonary recipients. PMID- 8222194 TI - Restrictive atrial septal communication as a determinant of outcome of cardiac transplantation for hypoplastic left heart syndrome. AB - BACKGROUND: Infant cardiac transplantation for hypoplastic left heart syndrome (HLHS) can be performed with a high success rate, but it remains unclear whether variations in anatomy or degree of illness before transplantation can affect ultimate survival. METHODS AND RESULTS: We analyzed the initial echocardiograms and pretransplant courses of 21 infants whose families chose transplantation as a primary therapy option for factors influencing survival. Sixteen of 21 were transplanted (mean wait, 18 days; range, 3 to 97 days) with 15 current (6- to 36 month) survivors, leading to a survival rate from presentation of 71% (70% confidence limit [CL], 61% to 81%) and an operative survival of 94% (70% CL, 81% to 99%). Seven of 20 listed for transplant (35%) had a severely restrictive interatrial communication, defined as a color flow jet width of 3 mm or less across the interatrial septum. Three of these 7 infants died at 4 (2) and 7 (1) weeks from congestive heart failure and progressive hypoxemia before transplantation. One died 2 months after transplant with an autopsy demonstrating pulmonary venous hypertensive disease, leading to a significantly higher relative risk for death (relative risk = 7.4, P = .01) for those infants with a severely restrictive interatrial communication versus those infants without severe restriction. The size and function of the right ventricle, left atrium, aorta, and tricuspid valve, as well as the magnitude of support on presentation, were not related to ultimate survival. CONCLUSIONS: A severely restrictive interatrial communication is a frequent component of HLHS. When primary cardiac transplantation is the chosen therapy for HLHS, it is a significant negative risk factor for death primarily before transplantation. PMID- 8222195 TI - Characterization and surgical ablation of acute atrial flutter following the Mustard procedure. A canine model. AB - BACKGROUND: A high incidence of refractory atrial flutter has been reported as a late postoperative complication of the Mustard procedure. The objective of this study was to reproduce experimentally the long-cycle-length atrial flutter that occurs in these patients and attempt to ablate the arrhythmia surgically. METHODS AND RESULTS: Nine dogs underwent a simulated Mustard procedure, which included a longitudinal right atriotomy, a septectomy, and a continuous suture line placed in the usual baffle location. All animals were subsequently inducible into sustained atrial flutter after surgery. Activation-time electrophysiological maps of the various pathways of atrial reentry were determined using atrial endocardial molds containing 252 bipolar electrodes. Four distinct reentrant circuits were observed in the following locations: (1) around the tricuspid value annulus (n = 5; cycle length, 175 +/- 5 milliseconds), (2) around the atriotomy incision (n = 2; cycle length, 208 +/- 3 milliseconds), (3) in a pathway involving both atria (n = 1; cycle length, 180 milliseconds), and (4) around the mitral valve (n = 1; cycle length, 135 milliseconds). A common pathway for eight of the nine reentrant circuits included the free-wall segment of the right atrium (excluding the mitral valve circuit). In the first three animals, surgical ablation of the atrial flutter was not attempted, but an incision made perpendicular to the atriotomy down to the tricuspid valve annulus resulted in termination of the atrial flutter in the other six animals. One of these animals was subsequently inducible into an atrial flutter of different morphology (cycle length, 125 milliseconds) that involved an isolated left atrial reentrant circuit. CONCLUSIONS: The Mustard procedure creates several anatomic substrates that increase the probability of the heart to develop atrial flutter, and these may be amenable to subsequent surgical correction. PMID- 8222196 TI - Excellent long-term results of cardiac valve replacement with the St Jude Medical valve prosthesis. AB - BACKGROUND: A retrospective review of the performance of the St Jude Medical valve prosthesis over a 12-year period was undertaken. METHODS AND RESULTS: Between 1978 and 1990, 696 St Jude Medical valve prostheses (351 isolated aortic, 191 isolated mitral, 64 double aortic/mitral) were implanted into 616 patients (mean age, 55 years). Concomitant coronary artery graft surgery was performed in 18%. During mid-1991, follow-up was conducted, yielding a 97% completion (16 lost), for a total of 3075 cumulative patient-years (mean, 5.0 years). Early (30 day) mortality rates were 3.1%, 5.2%, and 6.4% after aortic, mitral, and double valve replacements, respectively. The 5-year actuarial survival rates were 94.1 +/- 1.3%, 85.8 +/- 2.7%, and 86.3 +/- 4.6% and those for 10 years were 89.6 +/- 1.9%, 72.9 +/- 6.1%, and 83.0 +/- 5.4%, respectively. There were no structural valve failures. Anticoagulant-related hemorrhage was the most common valve related complication (28 events, 0.9% per patient-year), with three being fatal. Thromboembolism (13 events) occurred at a rate of 0.5% per patient-year (0.6% aortic, 0.3% mitral, 0.3% double). All of the nonstructural deteriorations (10 events, 0.3% per patient-year) were paravalvular leaks, including 3 aortic valve replacements in patients who developed hemolytic anemia. There was 1 valve thrombosis (0.03% per patient-year). Patients undergoing coronary artery graft surgery had lower survival and higher complication rates than patients without coronary artery graft surgery. There was a significant improvement (P < .001) in New York Heart Association functional class for the entire patient population. CONCLUSIONS: The St Jude Medical valve prosthesis has performed well in all positions over a 12-year period, with an acceptably low incidence of valve related complications. PMID- 8222197 TI - Randomized, prospective assessment of bioprosthetic valve durability. Hancock versus Carpentier-Edwards valves. AB - BACKGROUND: Although the major limitation of porcine valves is their finite durability, no controlled clinical data exist regarding the relative durability of the two porcine bioprostheses implanted most commonly today, the Carpentier Edwards (C-E) and Medtronic Hancock I (H) valves. METHODS AND RESULTS: To assess this question, 174 patients undergoing aortic (AVR) or mitral (MVR) valve replacement with a bioprosthesis between March 1980 and March 1982 were randomized to receive either a C-E or a H valve. There were 102 AVRs (54 C-E and 48 H) and 74 MVRs (39 C-E and 35 H). For both the AVR and MVR cohorts, the average patient age was 58 +/- 14 years (+/- SD). The male/female ratio was 2.2:1 for AVR and 0.57:1 for MVR. Clinical follow-up was undertaken periodically; the most recent follow-up closing interval was July through October 1992, and current follow-up was 96% complete. Cumulative follow-up totaled 1369 patient-years (mean, 7.7 +/- 3.6 years; median, 9.1 years; maximum, 12.0 years). The main focus of this analysis was bioprosthetic durability, using the AATS/STS guidelines defining "Structural Valve Deterioration" (SVD). Multivariate analysis revealed that (younger) age was the only significant (P = .024) independent predictor of SVD. Valve manufacturer (C-E versus H) and valve site (aortic versus mitral) did not emerge as significant independent risk factors for SVD. Actuarial rates (Cutler-Ederer) expressed as percent free of SVD (+/- SEM) at 10 years (n = number of patients remaining at risk) were 71 +/- 7% and 59 +/- 9% for the C-E (n = 26) and H (n = 17) groups, respectively, for the AVR cohort; for the MVR cohort, these estimates were 60 +/- 10% (n = 12) and 72 +/- 10% (n = 11), respectively, but these differences were not statistically significant (P = NS, Lee-Desu). CONCLUSIONS: After 10 years, there was no statistically significant difference in durability or other valve-related complications between the H and C E aortic or mitral valves. Based on current information, the choice of a porcine bioprosthesis should be based on factors other than durability, including ease of implantation, hemodynamic performance, and cost. PMID- 8222198 TI - Prognosis and management of anterolateral myocardial infarction in patients with severe left main disease and cardiogenic shock. The left main shock syndrome. AB - BACKGROUND: To identify the determinants of survival in patients with severe (> 75%) stenosis of the left main coronary artery (LM) and an acute (48 hours) anterolateral myocardial infarction (AAMI), we retrospectively analyzed the course of 34 such patients who presented to our institution over the last decade. METHODS AND RESULTS: LM disease was diagnosed arteriographically at presentation, and AAMI was determined by ECG, enzymatic, and kinetic criteria. Of the nine patients (26%) managed medically, seven patients (78%) were in cardiogenic shock (cardiac index < 2.0, left ventricular end-diastolic pressure > 25, and pulmonary edema), and all seven died in hospital. Twenty-five (74%) of the 34 patients were managed surgically or with angioplasty. Nine of these patients, of whom eight were in cardiogenic shock, also died in hospital. Regardless of the method of treatment, the presence of cardiogenic shock in this population was reproducibly a grave prognostic indicator. That is, 15 (94%) of the 16 patients in cardiogenic shock at presentation died in hospital, and only 1 (5%) of the 18 patients without cardiogenic shock died (P < .001). CONCLUSIONS: Thus, we propose that, because patients presenting with AAMI, severe LM stenosis, and cardiogenic shock (left main shock syndrome) have such a grave prognosis regardless of management, conservative measures may be indicated. PMID- 8222199 TI - Survival at 15 to 18 years after coronary bypass surgery for angina in women. AB - BACKGROUND: Coronary bypass surgery in women is associated with lower survival than in men. We need to know whether this is because of patient-related factors and whether the lower survival is present in all subgroups of patients and for all time periods during which the surgery was performed. METHODS AND RESULTS: Using actuarial techniques, we determined the outcome of coronary bypass surgery performed for chronic stable and unstable angina in 1979 women and 6927 men. The operative mortality was 2.7% for women and 1.9% for men (P = .02). The higher operative mortality in women was seen in those with three-vessel disease or greater and abnormal left ventricular function (5.4% versus 2.8%, P = .009) and those with stable angina (2.6% versus 1.5%, P = .006). The 5-, 10-, 15-, and 18 year survival for women was 86 +/- 0.9%, 70 +/- 1.5%, 50 +/- 2.5%, and 37 +/- 6.4%, respectively, and for men, 88 +/- 0.4%, 73 +/- 0.7%, 54 +/- 1.2%, and 42 +/ 1.9%, respectively (P = .03). The lower survival in women compared with men was seen in those with three-vessel disease or greater and abnormal left ventricular function (at 10 years, 69 +/- 1.8% versus 73 +/- 0.8%, P = .005). At 15 years, the incidence of reoperation was 26 +/- 2.4% versus 28 +/- 1.2% and of myocardial infarction, 30 +/- 2.8% versus 32 +/- 1.3%, P = NS for either. The incidence of no angina or mild angina was 70% in women and 78% in men, P < .0001. The operative mortality and late survival of those operated on in different time periods for either women or men was not significantly different. Women were older (64 +/- 9.4 versus 61 +/- 9.9 years, P < .0001) and smaller (body surface area, 2.0 +/- 0.2 versus 1.7 +/- 0.2 m2, P < .0001), had a higher incidence of diabetes, systemic hypertension, and unstable angina, and had a smaller lumen of the left anterior descending coronary artery (1.7 +/- 0.4 versus 1.9 +/- 0.4 mm, P < .0001), right coronary artery, and diagonal arteries. More men were smokers, and men had a higher incidence of prior myocardial infarction, previous coronary bypass surgery, and extent of coronary disease and of abnormal left ventricular function. The Cox regression model of survival showed that independent risk factors for lower survival were older age, previous coronary bypass surgery, previous myocardial infarction, and diabetes. Sex was not an independent risk factor for poorer survival. CONCLUSIONS: Women have a higher operative mortality and lower long-term survival than men after coronary bypass surgery for angina. However, the differences are small, even if statistically significant. Importantly, patient-related factors and not sex are independent predictors of poorer survival. Therefore, coronary bypass surgery should not be delayed or denied to women who have the usual indications for surgery. PMID- 8222200 TI - Changing clinical characteristics of coronary surgery patients. Differences between men and women. AB - BACKGROUND: There is uncertainty as to the relative efficacy of coronary artery bypass graft surgery (CABG) in men and women. Differences between sexes may be of greater importance than heretofore because of increased frequency of risk factors for adverse outcome. It is the purpose of this study to examine the changing clinical characteristics of patients undergoing CABG and to determine the influence of these changes on the results of CABG in men and in women. METHODS AND RESULTS: The source of data was the clinical database at Emory University Hospitals. The population comprised 13,368 patients (2648 women, 10,720 men) who had cardiac catheterization followed by CABG between 1974 and 1991. The patients were divided into the time periods 1974 through 1979 (532 women, 2792 men), 1980 through 1983 (727 women, 3151 men), 1984 through 1987 (695 women, 2619 men), and 1988 through 1991 (694 women, 2158 men). Data were collected prospectively and entered into a computerized database. The women were older (63 +/- 10 versus 58 +/- 10 years, P < .0001) and more frequently had diabetes (26.0% versus 15.4%, P < .0001). The in-hospital mortality was 3.8% in women and 1.6% in men. The multivariate correlates of death were older age, emergent surgery, reduced ejection fraction, female sex, diabetes, and more severe distribution of coronary arterial narrowings. Each of these risk factors for death became more prevalent, such that the mortality in men increased from 1.0% in 1974 through 1979 to 2.7% in 1988 through 1991, whereas the mortality in women increased from 1.3% in 1974 through 1979 to 5.4% in 1988 through 1991. CONCLUSIONS: The population undergoing CABG has aged, with correlates of in-hospital death becoming more common. The higher mortality in women could only partially be explained by other clinical variables. In addition, the risk of death after CABG has increased in women more than in men because women have consistently been older, have more emergent surgery and diabetes, and in recent years more women undergoing CABG have three vessel or left main disease. PMID- 8222201 TI - Venous coronary artery bypass surgery. A 15-year follow-up study. AB - BACKGROUND: Although the long-term results of isolated venous coronary artery bypass surgery are well known, there are few multivariate statistical data on such patient groups. METHODS AND RESULTS: We report on 428 consecutive patients, 383 men and 45 women with a mean age of 52.6 years, who underwent isolated venous aortocoronary bypass graft surgery with or without left ventricular aneurysm surgery between April 1, 1976, and April 1, 1977, and whom we followed prospectively. A multivariate analysis using the Cox regression model was performed to establish the determinants of long-term outcome. The hospital mortality and myocardial infarction rates were 3% and 6.3%, respectively. Complete revascularization was obtained in 77.6%. Follow-up was 99.8% complete and averaged 13.4 years (range, 1.5 months to 16.6 years). Actuarial survival after 5, 10, and 15 years is 94.2%, 82.4%, and 63%, respectively. The cumulative probability of event-free survival for cardiac death, acute myocardial infarction, reintervention, and angina pectoris at 5, 10, and 15 years, respectively, are 97.8%, 90.1%, 74.4%; 98.5%, 89.0%, 77.4%; 97.0%, 83.0%, 62.1%; and 77.8%, 52.1%, 26.8%. Left ventricular function and the number of vessels diseased are the independent preprocedural predictors of cardiac survival. Obesity and hypertriglyceridemia are preprocedural predictors of late myocardial infarction. Preoperative validity (Canadian Cardiovascular Society) and the number of diseased vessels are the predictors of recurrent angina. CONCLUSIONS: We conclude that the long-term results of isolated venous bypass graft surgery are dependent not only on well-known preprocedural factors such as number of vessels diseased, left ventricular function, and age but also on obesity and hypertriglyceridemia. PMID- 8222202 TI - Factors influencing 1-year patency of coronary artery saphenous vein grafts. Studio Indobufene nel Bypass Aortocoronarico (SINBA). AB - BACKGROUND: To evaluate the possible influence of a series of clinical angiographic and surgical variables on the 1-year patency of saphenous vein (SV) coronary graft, data collected prospectively in a multicenter randomized clinical trial were analyzed. METHODS AND RESULTS: The study group included 349 patients- 847 SV distal anastomoses--who underwent angiography at a median time of 374 days after surgery. By logistic binomial regression analysis, age, sex, smoking habits, hypertension, high cholesterol, previous myocardial infarction, and angina were not found to be significant factors leading to graft occlusion. Among the angiographic and surgical variables, the following were retained as predictive of higher occlusion risk: (1) vessel diameter (< or = 1.5 mm versus > 1.5 mm, odds ratio (OR) = 2.46); (2) the location of the grafted vessel, namely, the right coronary artery versus the left anterior descending (OR = 2.15); and (3) the wall motion of the vessel-dependent myocardial region (altered versus normal: OR = 2.12). The presence of two or three risk factors multiplied the occlusion risk up to 11-fold. CONCLUSIONS: The study suggests that vessel diameter, wall motion of the vessel-dependent myocardial region, and location of the grafted vessel are the main determinants of SV coronary graft patency during the first postoperative year. Knowledge of these artery-specific factors may provide a basis for estimating the risk of graft occlusion, thereby modifying surgical strategy and postoperative surveillance. PMID- 8222203 TI - Prognostic value of left ventricular ejection fraction response to exercise during long-term follow-up after coronary artery bypass graft surgery. AB - BACKGROUND: Recent data, albeit based on operations often performed more than 15 years ago, suggest that survival benefits of coronary artery bypass graft surgery (CABG) generally are lost within 10 years after operation. A reliable noninvasive method for periodic assessment of ischemia severity after operation might permit optimal timing of additional therapy to minimize loss of benefits. METHODS AND RESULTS: To determine the impact of left ventricular ejection fraction (LVEF) during exercise on prognosis in patients who have undergone CABG, results of rest and exercise radionuclide cineangiography were correlated with mortality, major nonsurgical cardiac events, and cardiac event-free or surgery-free survival in 192 patients who underwent index radionuclide study > or = 1 month (average, 26 months) after CABG. Average follow-up after study was 8.7 years among event-free patients. Initial events during follow-up included 31 deaths, 19 nonfatal myocardial infarctions, and 33 late repeat CABG or angioplasties (PTCA). Stepwise Cox regression analysis identified change (delta) in LVEF with exercise as the strongest independent predictor of cardiac death, major nonsurgical cardiac events, and cardiac event-free or surgery-free survival (P < .0001, all outcomes). Change in heart rate with exercise, completeness of revascularization, and New York Heart Association functional class for angina provided additional independent information. With each 10% decrement in delta LVEF, the hazard of cardiac death increased more than twofold, and the hazard of major nonsurgical cardiac events considered alone or in combination with repeat CABG or PTCA increased > 1.5 times. CONCLUSIONS: Thus, assessment of delta LVEF is prognostically useful after CABG. Assessment of this variable may help determine the need for repeat CABG. The utility of this approach now should be confirmed by longitudinal prognostic study. PMID- 8222204 TI - The ontogeny of estrogen receptors in heterochronic hippocampal and neocortical transplants demonstrates an intrinsic developmental program. AB - We investigated the intrinsic vs. environmental regulation of estrogen receptor (ER) ontogeny in the neocortex, hippocampus and hypothalamus by employing a heterochronic transplantation paradigm. These studies were based on previous reports demonstrating that neural ER develop asynchronously with quantitatively distinct ontogenetic profiles in various brain regions. Fetal (E14-15) hippocampal, frontal cortical or hypothalamic preoptic area (HPOA) primordial tissue was grafted into frontal cortical lesion cavities made in newborn (PND-0) rats. Thus, the grafted tissue was 1 week younger than the host. Two and 4 weeks following transplantation surgery, which corresponds to a theoretical donor age of PND-7 and PND-21, the grafts, a region of the host neocortex surrounding the transplant, and the host hippocampus, frontal cortex or HPOA (depending on graft type) were assayed for ER content using in vitro binding assays. ER concentration in hippocampal grafts at theoretical age PND-7 were significantly higher than those found in the host (PND-14) hippocampus and in the host neocortex adjacent to the transplant. By theoretical graft age PND-21, ER concentration in hippocampal transplants had decreased to levels comparable to those found in the host. This developmental pattern is analogous to that previously reported for the in situ hippocampus. A similar profile of ER concentration corresponding to the donor age developmental timetable was observed in neocortical grafts. ER levels in HPOA grafts did not change from theoretical donor age PND-7 to PND-21, which also corresponds to the normal ontogenetic profile. These data suggest that region-specific developmental patterns of ER expression in the rat brain are specified by embryonic day 14. PMID- 8222205 TI - Autoradiographic localisation of NMDA binding sites in brainstem cardiorespiratory areas of adult and newborn cats. AB - Quantitative autoradiography was used to determine the distribution of [3H]1-[1 (2-thienyl)cyclohexyl] piperidine ([3H]TCP) binding sites in the brainstem in order to localize the possible targets activated by excitatory amino acids in adult and newborn cats during autonomic rhythmic functions. Medium to high densities of binding sites were found in the nucleus tractus solitarius complex and the laterodorsal part of the pontine tegmentum. In the kitten, there was an apparent higher density in cranial motor nuclei proximal to these structures. It is concluded that there is an overlapping between the high density of NMDA receptors and the localization of cardiorespiratory neurons in cat as well as in kitten. PMID- 8222206 TI - Isoforms of agrin are widely expressed in the developing rat and may function as protease inhibitors. AB - The agrin family of extracellular matrix proteins may be important in the formation of the neuromuscular junction. Using in situ hybridization with a probe recognizing all agrin isoforms, we demonstrate that it is widely expressed during mammalian embryogenesis. In the developing rat, particularly high levels of expression are found in the dorsal root and cranial ganglia, gut, whisker rudiments, penis, snout, teeth, retina, hippocampus, cerebral cortex and the lining of brain ventricles. Functional analysis of the recombinant rat protein shows that it is a potent inhibitor of the proteases trypsin, chymotrypsin and plasmin but not thrombin or the plasminogen activators. We conclude that agrin and its isoforms may play multiple roles in mammalian development including the regulation of proteolysis in the extracellular matrix. PMID- 8222207 TI - Potent cocaine analogs inhibit [3H]dopamine uptake in rat mesencephalic cells in primary cultures: pharmacological selectivity of embryonic cocaine sites. AB - The cellular localization of the cocaine binding sites in primary cultures of embryonic rat mesencephalic cells was previously reported to differ from that observed in adult rat brain. In order to know whether this different localization was associated with a different pharmacological selectivity, we tested the effect of new cocaine analogs on tritiated dopamine ([3H]DA) uptake in primary cultures of rat embryonic mesencephalic cells. In these cultures, [3H]DA was taken up by a nomifensine-sensitive, but desipramine and fluoxetine-insensitive process, reflecting selective uptake by the dopaminergic transporter. 3 beta-(4 Chlorophenyl)tropan-2 beta-carboxylic acid methyl ester (RTI-COC-31) was by far the most potent inhibitor of the [3H]DA uptake, presenting an IC50 of 3.8 nM, while the corresponding analog with an unsubstituted phenyl ring (WIN 35,065-2) was 38 times less potent. The enantiomer of WIN 35,065-2, namely WIN 35,065-3, was 30 times less potent than the former. A similar pattern was found for the relative ability of these compounds to inhibit binding of the radiolabeled cocaine derivative [125I]RTI-55 to membranes prepared from mesencephalic cultures. The order of potencies found for the three cocaine analogs on mesencephalic cultures was similar to that previously obtained in [3H] WIN 35,428 binding experiments and [3H]DA uptake inhibition in adult rat striatum, suggesting that the pharmacological selectivity of cocaine sites functionally related to the DA transporter in cultured embryonic neurons does not differ from that obtained in adult rat brain. PMID- 8222208 TI - Reduction in the percentage of serotoninergic axons making synapses during the development of the superficial layers of the hamster's superior colliculus. AB - Immunocytochemistry with an antibody directed against a serotonin (5-HT)-bovine serum albumin conjugate was combined with electron microscopy to assess the synaptic organization of the serotoninergic projection to the stratum griseum superficiale (SGS) of the superior colliculus of hamsters killed on postnatal days (P) 0, 3, 7, 12, 15, 20 and > 60. At least 200 5-HT-immunoreactive profiles were examined at each of these ages. In the newborn (P0) animals, 36.6% of the 5 HT-positive profiles made conventional synapses. This percentage remained fairly constant until P15 when only 17.3% of the profiles made conventional synaptic contacts. On P20, this value decreased to 10.5% and in the adult animals, it fell to 4.4%. These results thus indicate a dramatic age-related change in the synaptic organization of the 5-HT input to the hamster's superior colliculus. PMID- 8222209 TI - Transient outgrowth from retinae implanted in the neonatal rat cerebral cortex. AB - Retinae placed in the cortex of neonatal rodents show substantial outgrowth, both along the subpial margin and within the cortical white matter, which is not sustained beyond 6 days post-transplantation. These results, in combination with previous studies, suggest that optic axon outgrowth is sustained for a finite period, after which it requires target-derived influences for its long-term maintenance. PMID- 8222210 TI - Acetylcholinesterase as an early marker of the differentiation of dorsal thalamus in embryonic rats. AB - The enzyme acetylcholinesterase (AChE) is transiently expressed in rats by neurons of the principal sensory thalamic nuclei, although these neurons do not use acetylcholine as a neurotransmitter. Reports that AChE expression begins at late embryonic stages led to the proposal that AChE may function in the establishment of connections, but not in earlier events. However, we find AChE reactivity in rat dorsal thalamus 5 days earlier than previously described. Cells that form the ventrobasal complex (VB), the dorsal lateral geniculate nucleus (dLG) and the medial geniculate nucleus, express AChE as they migrate and aggregate into definitive nuclei. AChE-positive cells are occasionally observed in the dorsal thalamic neuroepithelium, but are more common in others regions of the diencephalic neuroepithelium. AChE reactivity delineates VB and dLG earlier than Nissl-stained cytoarchitecture. These findings indicate that AChE is an early marker of neuronal differentiation. Certain properties of AChE, together with its early detection, are consistent with a proposed role in the migration of principal sensory neurons and their organization into discrete nuclei. PMID- 8222211 TI - Ontogeny of prodynorphin gene expression in the rat hypothalamus. AB - Opioid peptides, deriving from prodynorphin, proenkephalin and proopiomelanocortin genes, have been shown to modulate brain development. Prodynorphin gene expression was studied here by in situ hybridization in the developing rat hypothalamus using oligodeoxynucleotide probes. Prodynorphin mRNA synthetizing cells were observed in the ventromedial hypothalamic nucleus, the supraoptic and the paraventricular nuclei from embryonic days 16, 18 and 21, respectively. We detected no transient expression of prodynorphin gene in the rat hypothalamus. Prodynorphin mRNA-containing cells were also observed prenatally in the striatum, the cortex, the hippocampus and the amygdala. When compared with data from the literature, our results suggest that translation may immediately follow transcription of prodynorphin gene in the supraoptic nucleus. The presence of prodynorphin mRNA in the developing rat hypothalamus also raises the possibility of an involvement of prodynorphin-derived peptides in developmental processes and/or in the maturation of adult neural regulations. PMID- 8222212 TI - Early embryonic quail dorsal root ganglia exhibit high affinity nerve growth factor binding and NGF responsiveness--absence of NGF receptors on migrating neural crest cells. AB - Dorsal root ganglia (DRG) midway through development require nerve growth factor (NGF) for survival and differentiation. These studies investigated when avian neural crest cells or DRG first exhibit high affinity NGF receptors in situ, and whether early embryonic cells expressing high affinity NGF receptors are responsive to NGF. Unfixed cryostat sections of quail embryos were exposed to varying concentrations of [125I]NGF to distinguish between high and low affinity binding. Radioautography revealed an absence of [125I]NGF binding on migrating neural crest cells in situ. Both high and low affinity NGF receptors were first detected in differentiating DRG at E3.5 (stage 23). The presence of high affinity receptors was additionally confirmed by identification of a high molecular weight complex on radioautographs of gels following cross-linking of [125I]NGF to dissociated DRG. The presence of high affinity NGF receptors in E3.5 DRG was unexpected since DRG have been reported to be unresponsive to NGF prior to the midpoint of development. Exposure of E3.5 DRG neuron-enriched cultures to exogenous NGF resulted in approximately 30% more neurons after 24 h in vitro. The effect of NGF was blocked by anti-NGF and was shown to be dose dependent. It remains to be determined whether the increase in cell number is due to a survival or mitogenic effect. PMID- 8222213 TI - Developmental expression of the alpha 6 GABAA receptor subunit mRNA occurs only after cerebellar granule cell migration. AB - Using a competitive polymerase chain reaction (PCR) and appropriate internal standards, we have analyzed absolute amounts of the alpha 6 GABAA receptor subunit mRNA in the postnatally developing cerebellum and neocortex. The PCR data have shown that absolute amounts of the alpha 6 receptor subunit mRNA in the cerebellum increase dramatically (nearly 100-fold) during the second postnatal week, reaching maximal levels by postnatal day 21 (1 fmol/microgram total RNA). The absolute amount of the alpha 6 GABAA receptor subunit mRNA in the cortex at postnatal day 1 was 2 amol/microgram total RNA and increased to 7 amol/micrograms total RNA by postnatal day 14. No further increase in alpha 6 mRNA expression in the adult cortex was observed. Microscopic analysis of emulsion coated and counterstained sections indicated that alpha 6 GABAA receptor subunit mRNA labeling was only detected in the internal granule cell layer and not in either the external granule cell layer or in migrating granule cells. The alpha 1 GABAA receptor subunit mRNA increased in the cerebellar cortex with a similar temporal profile, although its distribution extended to additional cell types (Purkinje cells, stellate/basket cells and possibly cerebellar astrocytes). The temporal expression of these two GABAA receptor subunit mRNAs is coincident with the formation of synaptic contacts in the granule cell dendrites suggesting that afferent pathways innervating these neurons following cell migration may play a critical role in increasing the expression of mRNAs encoding the alpha 1 and alpha 6 GABAA receptor subunits. PMID- 8222214 TI - Simultaneous determination of vitamins A and E and carotenoids in plasma by reversed-phase HPLC in elderly and younger subjects. AB - A reversed-phase high-performance liquid-chromatographic method for the simultaneous determination of retinol, alpha-tocopherol, alpha-carotene, beta carotene, cryptoxanthin, lutein/zeaxanthin, and lycopene is described. This method was applied to plasma measurements in healthy young and elderly subjects. The plasma, deproteinized with ethanol, is extracted twice with n-hexane. After evaporation, the residue is dissolved in 50 microL of tetrahydrofuran and made up to 200 microL with ethanol. Samples (50 microL) are injected onto a 250 x 4.6 mm column of 5-microns-particle Spherisorb ODS1 (Phase Separations) that had been equilibrated with solvent mixture A:B (90:10 by vol) [A = 100 mmol/L ammonium acetate in methanol: acetonitrile (80:20 by vol) and B = 100 mmol/L ammonium acetate in water] at 2 mL/min. The analytes are eluted by running a 12-min linear gradient to 100% A; solvent A is then maintained for 10 min. Intrabatch CVs were 2.3%, 3.3%, 2.8%, 3.6%, 3.6%, and 3.0% for retinol, alpha-tocopherol, lutein/zeaxanthin, cryptoxanthin, lycopene, and beta-carotene, respectively. The corresponding interbatch CVs were 4.9%, 5.8%, 12.3%, 6.5%, 8.0%, and 3.4%. PMID- 8222215 TI - Evaluation of EMIT Cyclosporine Assay for use with whole blood. AB - We evaluated the EMIT Cyclosporine Assay, designed for specific quantification of cyclosporine (CsA) in whole blood. The assay was run on the Roche Cobas Mira or Cobas Mira S analyzer. Analytical recovery from both normal donor whole blood and transplant patients' samples was within 17% of the standards. Measurement of diluted out-of-range samples also yielded excellent recovery (101-105%). Within run, between-run, and total CVs were < or = 8.1%, 10.9%, and 12.1%, respectively. The detection limit was < 32 micrograms/L. The assay was linear from 0 to 500 micrograms/L. No significant cross-reactivity was observed for CsA metabolites AM1, AM19, and AM4N. Slight cross-reactivity occurred with metabolite AM9; 670 micrograms/L AM9 increased the measured CsA concentration by 49 micrograms/L. High concentrations of bilirubin, uric acid, triglycerides, and cholesterol, as well as 54 commonly coadministered drugs did not interfere with CsA quantification. Similarly, neither extreme values of hematocrit nor choice of anticoagulant affected CsA recovery. Sample extract stability was > 4 h, and assay calibration was stable for at least 2 weeks. Patients' samples analyzed by the EMIT assay showed strong correlation with both HPLC and 125I-RIA (r > 0.97). We conclude that the EMIT Cyclosporine Assay provides a convenient, accurate, and precise method for specific quantification of CsA in whole blood. PMID- 8222216 TI - HPLC assay of melatonin in plasma with fluorescence detection. AB - We report an HPLC assay for melatonin that incorporates automated injection, methanol/water mobile phase, and fluorescence detection. Plasma samples were extracted by solid and liquid phases. Recovery was > 70% for 1-10 mL of plasma extracted, approximately 40 pg-250 ng of melatonin. Samples were dried and reconstituted in 100 mL/L methanol. Injections were 25 microL or 150 microL, depending on sample concentration, and the melatonin peak was eluted in 380 mL/L methanol. The detection limit of the assay was 6 pg on the column, allowing a practical sensitivity in plasma of 11 pmol/L for 8-mL samples and 34 pmol/L for 2 mL samples. More than 100 plasma samples from volunteers and patients were assayed and the results compared with an established RIA. The mean daytime concentration of melatonin was 20.7 pmol/L (SEM = 1.2) and 18.5 pmol/L (SEM = 1.6) for HPLC and RIA, respectively, and the mean nighttime concentration was 82.4 pmol/L (SEM = 6.5) and 82.2 (SEM = 7.3), respectively. PMID- 8222217 TI - Artificial neural networks in diagnosis of thyroid function from in vitro laboratory tests. AB - We studied the potential benefit of using artificial neural networks (ANNs) for the diagnosis of thyroid function. We examined two types of ANN architecture and assessed their robustness in the face of diagnostic noise. The thyroid function data we used had previously been studied by multivariate statistical methods and a variety of pattern-recognition techniques. The total data set comprised 392 cases that had been classified according to both thyroid function and 19 clinical categories. All cases had a complete set of results of six laboratory tests (total thyroxine, free thyroxine, triiodothyronine, triiodothyronine uptake test, thyrotropin, and thyroxine-binding globulin). This data set was divided into subsets used for training the networks and for testing their performance; the test subsets contained various proportions of cases with diagnostic noise to mimic real-life diagnostic situations. The networks studied were a multilayer perceptron trained by back-propagation, and a learning vector quantization network. The training data subsets were selected according to two strategies: either training data based on cases with extreme values for the laboratory tests with randomly selected nonextreme cases added, or training cases from very pure functional groups. Both network architectures were efficient irrespective of the type of training data. The correct allocation of cases in test data subsets was 96.4-99.7% when extreme values were used for training and 92.7-98.8% when only pure cases were used. PMID- 8222218 TI - Synthetic peptide-based immunoassay for amino-terminal propeptide of type I procollagen: application for evaluation of bone formation. AB - Serum biochemical markers are powerful tools for the evaluation of bone turnover. In this study, we developed a radioimmunoassay, using a synthetic peptide for the N-terminal fragment of human type I [alpha 1(I)] procollagen (N-PCP). A 14-amino acid peptide was synthesized from the amino terminus and used to generate antibodies in rabbits. The synthetic peptide was used as standard and tracer in the assay. Both native type I amino procollagen (PINP), which was purified from skin fibroblasts, and human serum displaced tracer binding in parallel with the synthetic peptide. The range for measurement of N-PCP in serum was 0.7 to 30 micrograms/L (0.21-9.18 nmol/L). In a sample of 17 normal adults and 13 children (ages 9-16 years) there was a strong correlation between serum N-PCP determined by this assay and both skeletal alkaline phosphatase isoenzyme and osteocalcin, markers of bone formation. Serum concentrations of N-PCP in a group of normal children were eightfold higher than concentrations in normal adults, with no overlap between the two groups. N-PCP also correlated with C-terminal type I procollagen determined with a commercially available kit (r = 0.92). PMID- 8222219 TI - Iron-deficiency anemia and thalassemia trait differentiated by simple hematological tests and serum iron concentrations. AB - The diagnostic value of common hematology tests in differentiating between thalassemia trait and iron-deficiency anemia (IDA) was assessed. Serum iron concentration was included in a second stage of the diagnostic model and its predictive contribution was determined. After applying discriminant analysis to the data, minimization of Wilks's lambda (lambda) criterion was used to select the best predictive variables. A training sample of 754 subjects previously classified as either IDA (428) or thalassemia trait (326) was used to determine the classification rule. When serum iron concentrations were included, the model showed a higher predictive capacity than that constructed from hematological variables only (D = 2.342 RBC - 0.079 Hb + 3.627 log[Fe] - 6.459, where D = discriminant score). The sensitivity, specificity, and diagnostic efficiency associated with this model, as assessed on a control sample of 256 patients (137 thalassemia and 119 IDA), were 94.2%, 91.6%, and 93.0%, respectively. This model, when compared with those of other authors, has the highest diagnostic efficiency. PMID- 8222220 TI - Fast lipoprotein chromatography: new method of analysis for plasma lipoproteins. AB - Fast lipoprotein chromatography (FLPC) is a novel method for quantifying lipoproteins. Plasma proteins are separated by fast-flow gel filtration. Lipoproteins are detected by post-column derivatization with an enzymatic cholesterol reagent. FLPC resolves very-low-, low-, and high-density lipoproteins (VLDL, LDL, and HDL, respectively) and completely separates apolipoprotein Al- and apolipoprotein B-containing lipoproteins. CVs for VLDL-cholesterol, LDL cholesterol, and HDL-cholesterol are 5.8%, 2.0%, and 1.9%, respectively. We compared FLPC with a combined ultracentrifugation and precipitation method and obtained correlation of r = 0.979, 0.978, and 0.933 for VLDL-cholesterol, LDL cholesterol, and HDL-cholesterol, respectively. Triglyceride concentrations up to 9.00 g/L did not interfere with the quantification of lipoproteins by FLPC. We conclude that FLPC is a precise and reliable method for the analysis of plasma lipoproteins that complements conventional techniques. PMID- 8222221 TI - Colorimetric solid-phase minisequencing assay illustrated by detection of alpha 1 antitrypsin Z mutation. AB - In solid-phase minisequencing, a defined point mutation is detected in microtiter plate-immobilized DNA by a single nucleotide primer extension reaction. We have here developed the method into a colorimetric assay and applied it to the detection of the Z mutation of the alpha 1-antitrypsin gene. We used novel nucleoside triphosphates modified with dinitrophenyl (DNP) hapten, permitting detection by anti-DNP-alkaline phosphatase conjugate, with p-nitrophenyl phosphate as substrate. The Z mutation is detected in two reactions: DNP-labeled dCTP is incorporated when the template is normal, DNP-dUTP when the Z mutation is present. Both modified nucleotides were incorporated with high specificity and with an efficiency similar to that of unmodified nucleotides. The test results are measured by spectrophotometry, yielding quantitative absorbance values. Calculation of the ratio of C to U signal permitted unambiguous distinction of normal homozygous, ZZ homozygous, and ZM heterozygous genotypes. The colorimetric minisequencing assay is rapid, standardized, and automatable, and thus provides an accurate and simple alternative for the analysis of known point mutations. PMID- 8222222 TI - Relation between serum and whole-blood ethanol concentrations. AB - An algorithm is suggested for interpretation of serum ethanol concentrations under legal statutes that specify whole-blood alcohol concentrations. The algorithm uses the distribution of individual serum:whole-blood alcohol concentration ratios to allow calculations at various levels of confidence that can be related to legal standards of evidence. Serum:whole-blood alcohol concentration ratios were determined for 211 patients. The ratios ranged from 0.88 to 1.59 (median 1.15). The distribution of ratios was positively skewed, but the logarithms of the ratios were normally distributed. This allowed the parametric calculation of a range of ratios of 0.90-1.49 for the central 99% of the population and a range of 0.95-1.40 for the central 95%. The serum:whole blood alcohol concentration ratio was independent of both the serum alcohol concentration (R2 = 0.005) and the hematocrit (R2 = 0.03). PMID- 8222223 TI - Kinetic parameters for the cleaved substrate, and enzyme and substrate stability, vary with the phosphoacceptor in alkaline phosphatase catalysis. AB - Nine different isoenzymes and (or) isoforms of alkaline phosphatase (ALP; EC 3.1.3.1) from human tissue were studied with respect to Km and Vmax values for p nitrophenyl phosphate (p-NPP) in seven different potential phosphoacceptors/buffers. Generally, the phosphoacceptors/buffers with the lowest affinity for p-NPP (highest Km values) gave the highest Vmax values; for the nine enzyme forms in this study, the mean Km and Vmax values were greatest in 2 (ethylamino) (EAE). The two amino-propanol buffers gave the lowest Km and Vmax values. The phosphoacceptors/buffers N-methyl-D-glucamine (MEG), diethanolamine, and Tris had intermediate Km and Vmax values. Hydrophilic liver ALP retained > 90% of its activity after 24 h at 30 degrees C in both 1.0 and 0.3 mol/L Tris and 2-amino-2-methyl-1,3-propanediol and in 0.3 mol/L MEG. This isoenzyme showed greatest inactivation upon prolonged exposure to 1.0 and 0.3 mol/L EAE, the activity at 24 h being approximately 50-66% of that at zero time. p-NPP underwent the greatest spontaneous degradation, approximately 2.5 times that of baseline levels, in 1 mol/L MEG. There was little degradation in all of the buffers tested at 0.3 mol/L or in Tris, EAE, and 2-amino-2-methyl-1-propanol at 1.0 mol/L. PMID- 8222224 TI - Method for determining reference changes from patients' serial data: example of cardiac enzymes. AB - Changes between serial laboratory test results can be significant, even if none of the individual results exceeds the reference interval. We developed a statistical method for the calculation of reference change limits from routine patients' data for situations in which the majority of the patients can be considered suitable reference subjects. The method was applied to cardiac enzyme data [creatine kinase (CK; EC 2.7.3.2), creatine kinase isoenzyme MB (CK-2), lactate dehydrogenase (LD; EC 1.1.1.28), and lactate dehydrogenase isoenzyme 1 (LD-1)] from 2029 consecutive patients. We used hospital discharge diagnoses to exclude patients with the diagnosis of myocardial infarction or myocarditis, but we also studied the characteristics of the method on unselected patients' data. The reference change limits derived from the diagnosis-selected patient group were as follows (U/L, activity measurements in serum at 37 degrees C according to Scandinavian recommendations): CK from -39 to 27, CK-2 from -8 to 7, LD from -86 to 85, and LD-1 from -19 to 15. Similar limits were obtained by conventional statistical methods from a group of 29 hospitalized patients with no myocardial symptoms. Our results suggest that it is possible to produce clinically applicable reference change limits from routine data. PMID- 8222225 TI - Intraindividual variation of glycohemoglobin: implications for interpretation and analytical goals. AB - Intraindividual variation (CVi) for glycohemoglobin (GHb) was estimated from serial measurements in patients with diabetes in either stable or variable clinical control. GHb determinations were performed by an affinity column procedure with an analytical imprecision of 4.9% (weighted average; GHb 8.2 14.7%). Within the groups of patients, both a short- (28-32 days) and long-term (approximately 85 days) sampling protocol was used. The derived CVi for each category was 4.2% (n = 16, stable, short-term), 7.1% (n = 23, stable, long-term), 5.1% (n = 13, variable, short-term), and 9.8% (n = 21, variable, long-term). The mean GHb within each category was similar (approximately 11%), and there was no statistically significant difference in GHb values between categories. The results establish that the CVi for GHb is affected by both clinical control and the sampling time interval. These findings have important implications for the estimation of significant differences between serial GHb measurements and the setting of appropriate analytical precision goals. PMID- 8222226 TI - Electrochemical assay system with single-use electrode strip for measuring lactate in whole blood. AB - We have developed an assay system for measuring lactate in whole blood, consisting of a single-use strip of an enzyme-coated electrode and a small meter. The electrode strip is made of three plastic films: a cover sheet, a spacer, and an insulation layer printed with electrodes that are coated with lactate oxidase (EC 1.1.3.x) and ferricyanide as an electron mediator. The meter measures the magnitude of the anodic current of the reduced mediator by the enzymatic reaction and displays the lactate concentration 60 s after a blood sample (5 microL) is applied. The calibration curve was linear up to 20 mmol/L, and the between-run CVs at three concentrations were 1.7-8.4%. Lactate concentrations determined by this method (y) in blood samples from healthy individuals before and after exercise agreed with the results obtained by the conventional enzymatic method (x): y = 0.97x - 0.3, Sy/x = 0.7. This assay provides a rapid and convenient test for measuring blood lactate concentrations. PMID- 8222227 TI - Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of urine: concentration of urinary proteins by precipitation with coomassie blue. PMID- 8222228 TI - Solid-phase extraction protocol for isolating retinol-d4 and retinol from plasma for parallel processing for epidemiological studies. AB - Solid-phase extraction permits the parallel processing of samples in large numbers. We have applied this technique to the isolation of retinol isotopomers from plasma of humans participating in a study of vitamin A stable isotope dilution. The isotopomers were analyzed by gas chromatography/mass spectrometry. The extraction involves the separation of retinol from its aqueous matrix with a C18 silica-based sorbent followed by removal of lipid contaminants with an aminopropyl silica-based sorbent. Overall recovery of retinol from plasma was 47.2% +/- 1.8%. Purity of the retinol isolated from plasma is comparable with that obtained with a single HPLC method. This method permits the preparation of 32 samples per day by one analyst. Elimination of the need for HPLC permits sample preparation in the field with a minimum of equipment and technical skill. PMID- 8222229 TI - Competitive solid-phase enzyme immunoassay for melatonin in human and rat serum and rat pineal gland. AB - We describe a solid-phase competitive enzyme immunoassay for determination of melatonin in serum. The detection limit of the assay is 1.0 fmol/well. Low cross reactivity of the antiserum with other indoles, parallel serum extract dilution and melatonin standard curves, good analytical recovery, and within- and between assay CVs of 6.4-14.4% provide validation of the assay. Linear regression analysis of melatonin concentrations measured with this assay (y) and with a commercial 3H RIA (x) in 88 sera yielded the relation y = 0.62 x - 0.76, Sy/x = 0.03. Values for melatonin in serum samples from healthy subjects are lower during the day than during the night. Melatonin response in rat serum and pineal gland to isoproterenol injection is similar to published RIA data. The analytical procedure is also simple. Thus, the assay should have practical applications in investigation of pineal function in both clinical and basic studies. PMID- 8222230 TI - HemoCue beta-glucose photometer evaluated for use in a neonatal intensive care unit. AB - We evaluated the HemoCue beta-Glucose Photometer system for use in our neonatal intensive care unit by assaying 178 heparinized whole-blood samples obtained by heel stick. The required sample size is 5 microL. Plasma glucose was analysed by a glucose oxidase/oxygen electrode methodology. Across the glucose range of 1.28 21.87 mmol/L, the regression slope was 0.976 (r = 0.976, Sy/x = 0.475). For samples with hematocrit < or = 0.30, the regression slope was 0.981 (r = 0.950, Sy/x = 0.415); for hematocrit of 0.31-0.49, the regression slope was 0.984 (r = 0.972, Sy/x = 0.508); and for hematocrit > or = 0.50, the regression slope was 0.959 (r = 0.988, Sy/x = 0.394). Human whole blood, bovine whole blood, and bovine serum-based quality-control materials were studied. Except for assays of the low-concentration human control material, the total CV was < 3.5%. The accuracy and precision of the HemoCue system were comparable with those of conventional laboratory instrumentation. PMID- 8222231 TI - Automated enzymatic analysis of inulin. AB - We have developed an automated enzymatic assay for quantitation of inulin in plasma and urine that can be performed on the Cobas FARA II. In the assay, inulinase hydrolyzes inulin to fructose, and sorbitol dehydrogenase converts fructose to sorbitol with consumption of NADH, which is detected by spectrophotometry. The method incorporates a sample blank (inactivated inulinase) for each specimen to subtract contributions of endogenous fructose. Recovery of fructose or inulin was near 100%, with linearity to 300 mg/L. The enzymatic assay (y) agreed well with an anthrone comparison method (x) for analysis of inulin in both urine specimens (y = 1.00x - 138; Sy/x = 714) and plasma specimens (y = 1.00x - 3.5; Sy/x = 5.5). Glucose at 300 mg/L yielded an apparent inulin value of 1.3 mg/L in the enzymatic assay, but reacted at nearly 10% equivalency in the anthrone assay. Interferences from sorbitol, mannitol, and xylitol were negligible. CVs for day-to-day precision studies were 1-4%. The automated enzymatic assay of inulin is faster and avoids the use of caustic reagents required by the classical anthrone method. PMID- 8222232 TI - Platelet serotonin content and free and total plasma tryptophan in healthy volunteers during 24 hours. AB - The determination of platelet serotonin (5-HT) and plasma tryptophan concentrations is useful in the diagnosis, investigation of etiologies, and treatment of psychiatric disorders. To determine the usual circadian variations in platelet 5-HT and free and total tryptophan concentrations, we measured these variables during 24 h at 1-h intervals and every 30 min from 2000 to 0800 in seven clinically healthy young men with an HPLC method. No common circadian rhythm for platelet serotonin concentrations was observed in our subjects; however, there was a distinct rhythm for both free and total plasma tryptophan: Concentrations were maximal in the afternoon and minimal during the night. PMID- 8222233 TI - Inaccurate measurement of 17 beta-estradiol in serum of female volunteers after oral administration of milligram amounts of micronized 17 beta-estradiol. PMID- 8222234 TI - Quality blood specimens (for potassium measurement) with a standardized finger incision device. PMID- 8222235 TI - A scorecard doesn't help when the players keep changing shirts. PMID- 8222236 TI - Conjugated, but not unconjugated, bilirubin negatively interferes in Hitachi 747 assay of inorganic phosphorus. PMID- 8222237 TI - Phospholipids in EDTA-treated plasma and serum. PMID- 8222238 TI - Urinary screen for acetaminophen (paracetamol) in the presence of N acetylcysteine. PMID- 8222239 TI - Microwave-induced rapid hydrolysis of acetaminophen and its conjugates in urine for emergency toxicological screen. PMID- 8222240 TI - Above-normal urinary excretion of albumin and retinol-binding protein in patients with acute myocardial infarction. PMID- 8222241 TI - Enzyme confusion. PMID- 8222242 TI - Lipemia interference in Beckman Diatrac hemoglobin A1c procedure removed. PMID- 8222243 TI - Immunochemical assay of transferrin and iron saturation in serum. PMID- 8222244 TI - Quantification of medrogestone in serum by isotope dilution mass spectrometry. PMID- 8222245 TI - Phenylketonuria mutant alleles in different populations: missense mutation in exon 7 of phenylalanine hydroxylase gene. PMID- 8222246 TI - International standardization of glycohemoglobin measurements: practical application. PMID- 8222247 TI - Novel mutation in exon 7 of phenylalanine hydroxylase gene in a Chinese patient with phenylketonuria. PMID- 8222248 TI - Models of neoplasia and their diagnostic implications: a historical perspective. AB - In comparison with normal cells, cancer cells have an enhanced ability to trap both nitrogen and energy; an enhanced operation of the glycolytic and direct oxidative pathways, leading to accumulation of lactate and increased production of NADPH; and a greater content of lysosomal hydrolases. These changes represent a reprogramming of gene expression, which, at its most specific, is accompanied by the reappearance in the cell and ultimately in the body fluids of oncodevelopmental proteins not normally found in mature adult tissues. The most florid stage of this reprogramming leads to the metastatic phenotype, which confers upon the cancer cell the ability to stimulate angiogenesis, invade the bloodstream and lymphatic vessel, and arrest and proliferate in distant tissues. The diagnostic implications of these phenotypic changes are illustrated for cancer of the cervix uteri and cancer of the colon. We also review the classical theories of neoplasia, including the cellular anoxia concept of Warburg, the deletion hypothesis of Potter, and various other mechanisms emphasizing genomic derepression and impaired immunity. The critical steps in chemical carcinogenesis are described, and the Vogelstein-Lane model is presented, emphasizing the stepwise and cumulative genomic changes affecting chromosomes 5q, 17p, 18q, and gene amplification of chromosome 12 as well as genomic instability resulting from reduced DNA methylation. The main consequences of these genomic alterations include overexpression or activation of oncogenes such as c-myc and k-ras, together with mutation or functional inactivation of suppressor genes such as p53. Finally, the implications of these findings for diagnosis and management are illustrated by reference to recent investigations in cancers of the breast, colon, and bladder, in which these genomic alterations can be detected by examination of appropriate cellular material and by detection in serum of antibodies to the p53 gene product. PMID- 8222249 TI - Genomic components of carcinogenesis. AB - Many of the genes encoding growth factors, growth factor receptors, enzymes, and other effector molecules that regulate normal cell growth are designated protooncogenes. Oncogenes, those genes associated with cellular transformation, differ from their protooncogenic progenitors by being mutated, overexpressed, or expressed at inappropriate times or locations in the cell. One of the activities of growth factors is to prime cells to undergo programmed cell death, which is characterized by a series of morphologic changes called apoptosis. In normal cells, specific mediators must be activated or suppressed to bypass programmed cell death. In tumor cells, either the pathways leading to apoptosis are not functional or the mediators that normally "rescue" cells from this fate are overexpressed or constitutively activated. In addition to the biochemical pathways that drive cell division, there are others that limit cell proliferation; these, designated tumor suppressors, anti-oncogenes, or recessive oncogenes, must be inactivated in normal cells to allow passage through the cell cycle and cell proliferation. In contrast to oncogenes, which are overexpressed or activated in tumors, tumor-suppressor genes are frequently inactivated in tumor cells, either by mutation or deletion. Thus, in normal cells a series of checks and balances must be overcome to allow initiation and continuation of cell division. In tumors, these processes are aberrant, resulting in increased rates of cell division, increases in the proportion of cells in the cell cycle, or increased survival of activated cells. Therefore, tumor cells frequently accumulate genomic alterations, which may result in the activation of a particular array of oncogenes, the inactivation of specific tumor-suppressor genes, and the bypassing of programmed cell death. Trials of antitumor agents that act by exploiting the overexpression of oncogenes in tumors and of the biochemical pathways by which they mediate cell proliferation are currently underway. PMID- 8222250 TI - Clinical utility and validation of emerging biochemical markers for mammary adenocarcinoma. AB - The clinical utilities of the emerging biochemical markers for mammary adenocarcinoma CA 15-3, CA 549, CA M26, CA M29, and MCA (mucin-like carcinoma associated antigen) were assessed by a formal rating according to six desirable marker characteristics. All five indicators similarly have good specificities but limited sensitivities. As a consequence, these markers mainly meet just two desirable criteria: their frequency and degree of expression reflect tumor burden and prognosis, and they may correlate with therapeutic results. The validation of these assay properties by clinical trials was evaluated by another rating system, designed to assess proband sample selection, restrictions on allowable observations, and choice of statistical descriptors. By these benchmarks, the estimates of the prior probabilities of test outcome (sensitivity and specificity) are reasonably definitive, but conclusive judgments about the posterior probabilities of test outcome ("predictive values") and about values and costs associated with testing are not possible. Three approaches to enhance the limited clinical utility of biochemical breast cancer markers are considered: shifts of the diagnostic decision threshold, marker panels, and sequential testing. However, none of these strategies improves the described performance characteristics. PMID- 8222251 TI - Clinical applications of serum and tissue markers in malignant disease: breast cancer as the paradigm. AB - Within the past few years, the measurement of serum and tissue markers, especially the latter, has assumed a more significant role influencing clinical decisions about treatment and follow-up of patients with malignant disease. Breast cancer is a useful paradigm to illustrate the types and importance of these various markers. Tissue markers, including nuclear grade, steroid hormone receptors, DNA index, ploidy, expression of oncogenes or tumor-suppressor genes, epidermal growth factors, cathepsin D, proliferating cell nuclear antigen (PCNA), Ki-67, p32, and others, may influence choices of initial treatment as well as adjuvant chemotherapy and (or) hormone administration. The serial measurement of serum markers, those currently available and those on the horizon, for example, may offer a way to monitor patients at risk for recurrent cancer. Although the current role of these markers may be controversial, as information about them is collected and refined, in the future perhaps a panel of such studies could be incorporated into forthcoming clinical staging systems for carcinoma of the breast and other malignancies to define both treatment and outcome. PMID- 8222252 TI - Circadian pharmacodynamics of anticancer therapies. AB - Circadian variation in drug metabolism and tissue sensitivity to drugs affects their activity and toxicity. A growing body of data suggests that therapy may be improved and toxicity reduced by administering antineoplastic agents at carefully selected times of the day. Here I briefly review molecular, cellular, and organismic time-keeping mechanisms as well as cytokinetic, pharmacokinetic, and pharmacodynamic data, which support the predictable and exploitable nonlinear dynamic relation between dose and effect that occurs each day. PMID- 8222253 TI - Therapeutic drug monitoring in cancer management. AB - Several anticancer drugs display characteristics that make them suitable candidates for therapeutic drug monitoring (TDM), including substantial pharmacokinetic variability and a narrow therapeutic index. However, concentration-effect relationships (pharmacodynamics) of most antineoplastic agents have not been well defined, thus limiting the widespread clinical application of TDM for cancer chemotherapy. Strategic incorporation of pharmacokinetic studies during phase I-III clinical trials should facilitate the identification of concentration-effect relationships and the definition of clinically useful levels of treatment intensity. We review representative clinical studies that have defined pharmacodynamic relationships for methotrexate, teniposide, etoposide, carboplatin, and mercaptopurine. Given that TDM has impacted positively on the clinical use of many drugs belonging to other therapeutic classes, and that pharmacodynamic correlations have been identified in several recent studies of anticancer drugs, we consider implementation of TDM a rational strategy for optimizing the use of selected antineoplastics. PMID- 8222254 TI - Serum tumor markers for patient monitoring: a case-oriented approach illustrated with carcinoembryonic antigen. AB - The effectiveness of patient monitoring with serum tumor markers is improved when the patient's pretreatment baseline is established, regular serial testing is performed, and a clinically significant change of the marker is determined on the basis of objective criteria. Attention must be directed to (a) ensuring long-term assay precision, (b) timing specimen collection to avoid misinterpretation of paradoxical increases (induced by chemotherapy, surgery, or radiation), (c) noting changes in the production or clearance mechanisms of the marker, and (d) recognizing the variability of site-response in patients who have multiple locations of metastatic disease. Clinical decisions to alter therapy should not be based only on tumor marker measurements. In the absence of other clinical data, a tumor marker abnormality should be confirmed with serial samples collected at a time interval dictated by the half-life of the marker. Serial tumor marker testing can improve cancer patient care, especially when effective salvage therapies are available. PMID- 8222255 TI - Reimbursement of tumor marker tests. AB - The diffusion of new technology is significantly affected by coverage and reimbursement decisions. A variety of agencies (public and private) make decisions as to which technologies or treatments will be covered and what the levels of reimbursement will be. Recently, these agencies' coverage decisions have tended to be strongly affected by whether a technology is cost raising or cost reducing. Historically, a common effect of medical innovation has been improved quality of health care but with a corresponding increase in cost of delivery. The Medicare program has significant influence on the coverage policies of public and private third-party payers. This influence is especially visible in coverage and reimbursement decisions for cancer-related diagnostic procedures and systems. Coverage for these procedures and systems is not widespread because payers have not been convinced of the clinical usefulness of the assays. The industry must take the responsibility of working through the issues surrounding coverage and reimbursement of cancer-related diagnostic procedures with the payers. The ability to successfully negotiate payment for assays requires knowledge of Medicare coverage policies and a grasp of the reimbursement system. PMID- 8222256 TI - The FDA's perspective on the evaluation of tumor marker tests. AB - Cancer marker tests are often proposed for three intended uses: screening, diagnosis, and monitoring. For each intended use, performance characteristics need to be well defined. The utility of a marker in a given setting depends heavily on two predominant performance characteristics--sensitivity and specificity. These parameters must be established with respect to the intended clinical use of the marker. The value of the marker in a particular situation also depends on the effectiveness of therapy for the malignancy. In reviewing a cancer marker test, the US Food and Drug Administration focuses on both the proposed intended use statement and the clinical utility of the marker. The sponsor is expected to provide specific claims data in support of the safety and effectiveness of the device through well-designed and -executed clinical studies. Several cancer markers are already available. In the future, new markers are anticipated that may greatly expand the range of usefulness in cancer diagnosis screening and monitoring. PMID- 8222257 TI - Perspectives on the future of cancer markers. AB - More fundamental understanding of cell growth regulation will provide novel approaches for detecting, preventing, and treating different cancers. Activation of protooncogenes or loss of tumor-suppressor genes can have both prognostic and therapeutic importance. In epithelial ovarian cancer, poor prognosis is associated with continued expression or overexpression of tyrosine kinase growth factor receptors p170EGFR, p165fms, and p185erbB-2. Over-expression of erbB-2 (HER-2/neu) by breast and ovarian cancers already permits effective targeting of antibodies and immunotoxins. Ultimately, molecular analysis of individual cancers will guide the application of specific therapies to inhibit activated oncogenes or restore the function of tumor-suppressor genes. Circulating growth factors, oncogene products, and tumor-associated antigens may provide markers for earlier detection of some cancers. In epithelial ovarian cancer, concentrations of CA 125 can be increased 1-5 years before clinical diagnosis, and approximately 50% of patients with stage I disease have had an abnormal CA 125 concentration. Combining CA 125 with two novel markers--OVX1 and M-CSF--has retrospectively detected > 98% of stage I ovarian cancers. Although the specificity of the three markers is insufficient for cost-effective screening, serum tests for them could prompt the performance of transvaginal sonography, substantially decreasing the number of sonograms required. Genetic markers in the germ line of patients at increased risk for certain cancers will almost certainly influence strategies for prevention or detection. In familial breast, and ovarian cancer, a locus on chromosome 17q tracks risk of cancer in a fraction of kindreds. How often germline abnormalities will be detected in patients with apparently sporadic cancer remains to be determined. PMID- 8222258 TI - The application of molecular biology techniques to the diagnosis of hyperlipidaemia and other risk factors for cardiovascular disease. PMID- 8222259 TI - Deregulated production of interleukin-4 (IL4) in autoimmune thyroid disease assayed with a new radioimmunoassay. AB - A sensitive, reproducible and specific radioimmunoassay for human interleukin-4 (IL4) has been developed. Using 125I-labeled IL4 and polyclonal rabbit antisera raised against recombinant human IL4, a competitive inhibition assay was developed which could detect 5 pg/ml of human IL4. Other interleukins, growth factors, hormones, peptides and lectins did not affect the assay. IL4 was measured in supernatants of culture media of stimulated human peripheral blood mononuclear cells (PBMC). Kinetics of IL4 production in PHA-stimulated PBMC from seven normal subjects revealed that the peak levels of IL4 were seen at 24 h and then declined. Peak IL4 levels in PHA stimulation of PBMC from untreated patients with autoimmune thyroid diseases (Graves' disease and chronic thyroiditis) were significantly higher than normal controls. However, after treatment, IL4 production decreased to normal. The present study demonstrates the usefulness of quantitating human IL4 produced by PBMC and that there exists a deregulated production of IL4 in autoimmune thyroid diseases. PMID- 8222260 TI - Determination of proteins by a reverse biuret method combined with the copper bathocuproine chelate reaction. AB - A method of protein determination has been developed which combines the biuret reaction and the copper(I)-bathocuproine chelate reaction. Protein in the specimen forms a Cu(2+)-protein chelate complex (biuret reaction) during the first step. Excess Cu2+ is reduced to Cu+ by ascrobic acid, allowing the Cu+ to form a Cu(+)-bathocuproine chelate complex during the second step. The amount of Cu(+)-bathocuproine chelate complex formed is inversely proportional to the protein concentration. The sensitivity (epsilon = 1.4 x 10(6) 1.mol-1.cm-1 against human albumin) of this method was higher than that of the original Lowry (9.8 x 10(5)), pyrogallol red (1.0 x 10(6)) and commercially available Coomassie Brilliant Blue G.250 methods (6.7 x 10(5)). The color intensities of human gamma globulin, human globulin (fractions IV-1 and IV-4), bovine albumin, egg albumin and horse gamma-globulin against human albumin (100%) ranged from 92 to 101%. The results obtained with the present method (y) correlated well with those determined by the biuret method (r = 0.998, y = 0.98 chi - 0.002, x = 1.31, y = 1.29 g/l) in 30 diluted sera. These results confirm that this assay is similar in sensitivity to the original Lowry method, is rapid and has similar reactivity to each of the various proteins in biological fluids. PMID- 8222261 TI - Variable expression of leukocyte cytosolic broad-specificity beta-glucosidase activity. AB - The cytosolic beta-glucosidase activity that is found in a variety of mammalian tissues has no clearly defined function. In vitro assay conditions under which the broad-specificity beta-glucosidase hydrolyzes glucocerebroside at a significant rate have not been described. Nonetheless, it has been suggested that this enzyme might play an accessory role with lysosomal glucocerebrosidase in catalyzing the hydrolysis of glucosylceramide. However, this hypothesis would require that activity of both enzymes be low in severe cases of Gaucher disease in which there are pathological accumulations of glucosylceramide in one or more of the affected organs, i.e. spleen, liver and bone marrow. Information is lacking regarding the normal range of cytosolic beta-glucosidase activity in humans. p-Nitrophenyl-beta-D-mannoside was found to be a potent inhibitor (Ki = 0.068 mM) of cytosolic beta-glucosidase. In parallel studies, the activity of glucocerebrosidase was found to be minimally affected by p-nitrophenyl-beta-D mannoside at concentrations as high as 2.5 mM. This information was used to design an assay system that would allow us to estimate glucocerebrosidase and cytosolic beta-glucosidase activities in extracts of human leukocytes. Average cytosolic beta-glucosidase activity with 4-heptyl-umbelliferyl-beta-D-glucoside as a substrate was 8.8 nmol/h/mg protein in leukocytes from 356 subjects (range, 0.2-28). Average leukocyte glucocerebrosidase specific activity was 16 nmol/h/mg protein (range 5.3-45.7). No correlation was observed between cytosolic beta glucosidase and glucocerebrosidase activity for control and Gaucher heterozygote populations (r = 0.19 and 0.25, respectively). The wide range of leukocyte cytosolic beta-glucosidase activity in individuals tested in this study indicates that a substantial proportion of the population may lack sufficient cytosolic beta-glucosidase activity to assist a defective lysosomal glucocerebrosidase in hydrolyzing glucosylceramide. PMID- 8222262 TI - Lipoprotein lipase mass and activity in severe hypertriglyceridemia. AB - To clarify the role of defective lipoprotein lipase (LPL) in hypertriglyceridemia, the LPL masses and LPL activities in post-heparin plasma (PHP) were studied in severe hypertriglyceridemias. The developed sandwich enzyme immunoassay for the LPL was sensitive from 0.5 to 20 ng/ml of LPL in human PHP. The plasma LPL mass increased by heparin injection (30 USP units/kg) and was found to positively correlate with LPL activity. The mean LPL activity from PHP of normal controls was 2,960 +/- 1,057 nmol/ml/h. The mean LPL masses from human pre- and 15-min post-heparin plasma from normal subjects were 25 +/- 5 ng/ml and 224 +/- 60 ng/ml, respectively. Thus the specific activity of LPL from PHP of normal controls was calculated to be 13.3 mumol FFA released/h/microgram LPL. Among hypertriglyceridemic patients with over 1,000 mg/dl of serum triglyceride, the incidence of patients with LPL masses less than -2 standard deviations (S.D.) of those of average normal control subjects was found to be 27%. Seventy percent of patients showed specific activities within + 2 S.D. of those of average control LPL, and 30% showed significantly low specific activities less than -2 S.D. despite the fact that LPL masses were not less than -2 S.D. of the average normal controls. These results suggest that the evaluation of LPL masses in PHP would be useful for finding functionally defective LPL in patients with hypertriglyceridemia, and that up to 30% severe hypertriglyceridemias may have functionally defective LPL. PMID- 8222263 TI - Decreased renal excretion of beta-hexosaminidase in adults with insulin-dependent diabetes mellitus and normal renal function. AB - The activities of three lysosomal hydrolases and creatinine levels were measured in the plasma and urine of 11 adults (mean age, 28.1 years) with insulin dependent diabetes mellitus and 14 non-diabetic controls (mean age, 27.9 years). All of the patients were free of diabetic complications and non exhibited microalbuminuria. Fractional enzyme excretion (FEE) values between the two groups of subjects were calculated and compared for the following enzymes: beta hexosaminidase (N-acetyl-glucosaminidase), beta-glucuronidase and alpha galactosidase. The FEE value was calculated as the ratio of enzyme clearance to creatinine clearance. Relative to the non-diabetic control group, the FEE value for beta-hexosaminidase was approximately 2-fold lower (P = 0.02) in the diabetic subjects (means, 0.424 vs. 0.242, respectively). The FEE values for beta glucuronidase and alpha-galactosidase were not significantly different (P > 0.4) between the diabetic and control groups. These easily measured biochemical parameters in blood and urine and the resultant FEE value for beta-hexosaminidase may provide a means of assessing subtle deteriorative changes in renal function which occur in the early stage of diabetes before the onset of clinically evident complications. PMID- 8222264 TI - Enzyme reactor for urinary acylcarnitines assay by reversed-phase high performance liquid chromatography. AB - An immobilized enzyme reactor, made up acylcarnitine hydrolase, carnitine dehydrogenase and diaphorase in sequence, was developed for the sensitive and selective determination of urinary free and individual acylcarnitines by a reversed-phase high-performance liquid chromatography. A 100-microliter urine sample was directly injected onto the TSKgel ODS 80Ts column and eluted by a step gradient procedure. The eluent was mixed with the substrate solution of beta-NAD+ (1.0 mmol/l), resazurin (25 mumol/l) and Tris acetate (0.2 mol/l, pH 9.0). The mixture was passed through the immobilized enzyme reactor at 40 degrees C. Acylcarnitines were hydrolyzed and the converted to rezorufin which was measured by monitoring the fluorescence intensity at lambda EX = 560 nm and lambda EM = 580 nm. Free, acetyl-, glutaryl-, propionyl-, butyryl-, isobutyryl-, valeryl- and isovalerylcarnitine were determined within 55 min with detection limits (< 1 mumol/l) and within-run and day-to-day imprecision (C.V. < 6%). Free, acetyl- and isobutyrylcarnitine were found in normal urine. On the other hand, propionylcarnitine was detected in the urine of children with propionic aciduria and methylmalonic aciduria and multiple acylcarnitines were found in the urine of children with glutaric aciduria (type II). PMID- 8222265 TI - Erythrocyte galactokinase assay with high performance liquid chromatography. AB - An assay for erythtocyte galactokinase based on high performance liquid chromatographic determination of galactose 1-phosphate (Gal-1-P) is described. The determination of Gal-1-P was applied to a post-column fluorometric detection of reducing sugars using arginine. This method is as sensitive and accurate as conventional radioisotopic methods, but needs no radioisotopic facilities. It requires only a small blood sample and is suitable as a follow-up test in neonatal screening. PMID- 8222266 TI - Cerebrospinal fluid ferritin in brain necrosis: evidence for local synthesis. AB - Ferritin was measured in cerebrospinal fluid (CSF) and serum of an unselected neurological population. An increase in CSF ferritin was found to be associated with pathological processes in which there was either necrosis or haemorrhage involving the brain. There was no correlation between the CSF and serum concentrations of ferritin in the reference population. Neither was there any correlation between CSF ferritin and CSF albumin in the reference population. After subarachnoid haemorrhage, intrathecal production of ferritin was found to occur since in some patients the concentration of ferritin in CSF was higher than that of homologous serum. Even in the reference population the concentration of ferritin found in the CSF was much higher than could be explained by passive transfer across the blood-CSF barrier. Therefore local synthesis of ferritin by brain cells occurs even under normal circumstances. PMID- 8222267 TI - Mono-, di- and triacylglycerol concentrations in human plasma: effects of heparin injection and of a high-fat meal. AB - A sensitive method has been developed to measure specific mono-, di- and triacylglycerol concentrations in human plasma, using thin-layer chromatography and enzymatic assay. The levels of partial acylglycerols in human plasma from fasting subjects were lower than previous reports had suggested and amounted to less than 3% of the total acylglycerols. After heparin injection the plasma monoacylglycerol concentration increased markedly (P < 0.01) while the triacylglycerol concentration decreased significantly (P < 0.001). The plasma diacylglycerol concentration did not change significantly although it increased as a percentage of the total (P < 0.05); after heparin partial acylglycerols accounted for more than 10% of the total. After a high-fat meal the plasma concentrations of di- and triacylglycerol increased approximately two-fold (P < 0.005) but no significant change was observed for mono-acylglycerol. The percentage contribution of partial acylglycerols was unchanged (2.6% fasting, 2.4% postprandially). PMID- 8222268 TI - Characteristics of membrane-bound 5'-nucleotidase on human prostasomes. AB - Examination of prostasomes, isolated from human seminal plasma, showed that there was very little remaining paranitrophenylphosphatase activity when assayed in the presence of 10 mmol/l of tartrate and 2 mmol/l of levamisole. Under these conditions it was possible to study the prostasome membrane-bound 5'-nucleotidase activity, which was unaffected by these two inhibitors. The activity was considered to be located at the external surface of the prostasome membrane and a 50-60% increase in activity was obtained by the addition of 0.05% Triton X-100. The prostasome membrane-linked 5'-nucleotidase readily hydrolysed 5'-AMP. Two other 5'-nucleoside monophosphates, 5'-IMP and 5'-GMP, were also hydrolysed, but more slowly; 2'- or 3'-AMP were practically not attacked. The prostasome membrane linked 5'-nucleotidase obeyed Michaelis-Menten kinetics. Apparent Km for 5'-AMP was 11.2 +/- 2.1 mumol/l and Vmax 64.7 +/- 11.4 nmol/mg protein/min. These figures were somewhat changed in presence of 0.05% Triton X-100, the Km value being reduced by 30% and the Vmax value increased by 60%. Adenosine 5' (alpha, beta methylene) diphosphate (100 mumol/l), Ni2+ (10 mmol/l) and concanavalin A (20 micrograms/ml) were all potent inhibitors of the prostasome membrane-linked 5'-nucleotidase. PMID- 8222269 TI - Hormone-sensitive lipase: quantitation of enzyme activity and mRNA level in small biopsies of human adipose tissue. PMID- 8222270 TI - Apolipoprotein E phenotypes and genotypes as determined by polymerase chain reaction using allele-specific oligonucleotide probes and the amplification refractory mutation system in children with insulin-dependent diabetes mellitus. AB - The frequency of apolipoprotein E (apo E) phenotypes and genotypes due to allelic variation at amino acids 112 and 158 was analysed in 50 children with type I diabetes. Phenotypes were determined by isoelectric focusing and genotypes by the technique of polymerase chain reaction using allele-specific oligonucleotide probes (PCR/ASO) and the amplification refractory mutation system (ARMS). Discrepancies between phenotypes and genotypes as assigned by PCR/ASO were observed in 12 (24%) cases and by ARMS in eight (16%) cases. Results revealed the apo E3/3 genotype, as assigned by ARMS, to be the most frequent one (70%), followed by apo E3/4 in 16%, apo E2/2 in 2%, apo E2/3 in 8%, apo E2/4 in 2% and apo E4/4 in 2% of the cases. Apo E3/4 genotype and phenotype were more frequently present in the children with type I diabetes as compared with the diabetic adults previously reported on. PMID- 8222271 TI - A novel lectin-based enzyme-linked immunosorbent assay for the measurement of IgA1 in serum and secretory IgA1 in secretions. AB - Jackfruit lectin (Jacalin) specifically binds IgA1 and secretory IgA1 (sIgA1). Jacalin was successfully purified, and used as the immobilized phase in novel noncompetitive enzyme-linked immunosorbent assays (ELISAs) for the measurement of IgA1 in serum and sIgA1 in saliva, using jacalin affinity purified material as standards. A non-competitive ELISA for the measurement of total sIgA was also developed using commercially available reagents. PMID- 8222272 TI - Intrathecal synthesis of IgM and IgA in neurological diseases: comparison of two formulae with isoelectric focusing. AB - The intrathecal IgM and IgA immune response was studied by detecting oligoclonal IgM and IgA bands in CSF and by determining both the IgM/IgA indices as well as the intrathecal production of IgM and IgA by the Reiber formula. A good correlation was found between the demonstration of oligoclonal IgM (r = 0.890) and IgA bands (r = 0.927) and the Reiber formula. Compared with the finding of oligoclonal IgM/IgA bands calculation of an intrathecal IgM/IgA synthesis by the Reiber formula was less sensitive (86%). In two out of 22 controls and in 12 of 19 patients with polyradiculitis IgM and IgA indices were elevated while the other evaluation methods were negative. Even though oligoclonal IgM and IgA bands are considered to be a useful diagnostic tool in detecting intrathecal synthesis of IgM and IgA, i.e. in meningoencephalitis or neuroborreliosis for clinical practice, the Reiber formula provides reliable results. Detection sensitivity of assays for CSF-IgM and -IgA should be at least 1.0 mg/l. PMID- 8222273 TI - Determination of acylcarnitines in urine of patients with inborn errors of metabolism using high-performance liquid chromatography after derivatization with 4'-bromophenacylbromide. AB - A high-performance liquid chromatographic method is presented for the determination of urinary acylcarnitines. After solid phase extraction on silica columns the acylcarnitines are converted to 4'-bromophenacyl esters with 4' bromophenacylbromide in the presence of N,N-diisopropylethylamine. Complete derivatization was achieved at 37 degrees C within 30 min. The 4'-bromophenacyl esters were separated by high-performance liquid chromatography on a Hypersil BDS C8 reversed-phase column with a binary gradient containing varying proportions of acetonitrile, water and 0.1 M triethylamine phosphate buffer. Essentially baseline separation was obtained with a standard mixture containing 4' bromophenacyl esters of carnitine and synthetic acylcarnitines of increasing chain length ranging from acetyl- to palmitoylcarnitine. The method was used to obtain urinary acylcarnitine profiles from patients with propionic, methylmalonic and isovaleric acidemia and with medium-chain and multiple acyl-CoA dehydrogenase deficiency. Quantification of the acylcarnitines was achieved using undecanoylcarnitine as internal standard. PMID- 8222275 TI - Release of beta-hexosaminidase isoenzymes in cultured human fibroblasts. AB - We have used enzyme immunoassay methods to study the intra- and extracellular isoenzyme pattern of beta-hexosaminidase (Hex) in human fibroblast cultures. The released activity into the medium during 24 h was about 10% of the intracellular activity. Intra- and extracellular ratio of Hex B to total Hex (Hex A plus Hex B) was about one-third. Estimation of the molecular weight of the released activity showed that it corresponded to 150 kDa, which is equal to the high molecular mass precursor forms of the enzyme. NH4Cl is known to disturb the intracellular transport of lysosomal enzymes and increase the secretion of newly synthesized precursor forms. Addition of NH4Cl even at 3 mM resulted in an increased release of total Hex, which was already noted within 24 h. We speculate that the increased concentration of plasma NH4+ in patients with liver disease interferes with the intracellular distribution pathway of the lysosomal enzymes and this could contribute to the increased content of lysosomal enzymes present in plasma from these patients. Tunicamycin (0.3-3.0 micrograms/l) increased the percentage of Hex B in the medium, whereas an increased release of Hex was noted only after 48 h. Tunicamycin is known to enhance the secretion of N-linked oligosaccharide free forms of lysosomal enzymes. Thus the oligosaccharide chains on alpha- and beta-subunits seem to be important for the normal formation process of the Hex A isoenzyme. PMID- 8222274 TI - Hyperphenylalaninemia and pterin metabolism in serum and erythrocytes. AB - The relationship between blood phenylalanine concentrations and serum and erythrocyte biopterin and neopterin concentrations was investigated in 20 phenylketonuric patients with different dietary compliance. At serum phenylalanine concentrations ranging from 43 to 1004 mumol/l, a good correlation was found with serum biopterin (r = 0.76, P < 0.001) and with red blood cell biopterin (r = 0.62, P < 0.001). A similar correlation was found between serum neopterin and phenylalanine (r = 0.60, P < 0.001). The correlation between red blood cell neopterin and serum phenylalanine was less evident, however (r = 0.47, P < 0.005). After oral loading with phenylalanine (100 mg/kg body weight), serum and red blood cell biopterin concentrations increased in patients with classical phenylketonuria as well as in one patient with dihydropteridine reductase deficiency in response to the induced acute hyperphenylalaninemia. One patient suffering from 6-pyruvoyl tetrahydropterin synthase deficiency was loaded orally with tetrahydrobiopterin (20 mg/kg body weight). The kinetics of administered cofactor confirmed its rapid absorption, with early increase of serum concentrations followed by its transport into the red blood cells. The half-life of biopterin was approximately 7 h in serum and 15 h in red blood cells. Because both values are less than the half-life of phenylalanine (20-30 h) in serum, biopterin measurement offers no advantage in monitoring dietary control in hyperphenylalaninemic patients. PMID- 8222276 TI - A 125I-radioimmunoassay for diethylstilbestrol in serum of patients with prostatic cancer treated with stilphostrol. AB - A new radioimmunoassay for determining diethylstilbestrol in serum using N-(4'-OH [3'-125I]iodophenethyl)-6-(4-O-diethylstilbestryl)-hex anamide as a radiotracer and a double antibody as a separation reagent is described. The radiotracer is prepared by synthesizing 6-(4-O-diethylstilbestryl)-hexanoic acid and coupling its succinimidyl ester with mono-[125I]tyramine in tetrahydrofuran (16 h, 20-22 degrees C). The standard curve is linear (semi-log transformation) and the assay is sensitive (< 0.022 pmol/tube), reproducible (intra- and interassay coefficient of variation values, 5.3 and 8.1%, respectively), and accurate (recovery values, 95-101%), with a non-specific binding less than 3.2%. Diethylstilbestrol concentrations measured in sera of nine patients with prostatic cancer by the proposed assay ranged from 0.170 to 2.517 mumol/l, which corresponded to an only three-fold dosage variation. In all cases tested, dosing was adequate to retain markers of prostatic cancer in serum within accepted limits; nevertheless, individualization of dosing may be necessary to minimize toxicity. PMID- 8222277 TI - Fetal intestinal and renal origins of trehalase activity in human amniotic fluid. AB - Intestinal and renal trehalase isozymes have been distinguished in normal human amniotic fluid on the basis of their membrane-bound character and isoelectric point (pI). The intestinal trehalase was mostly membrane bound in amniotic fluid and had a pI around 4.60. In contrast, the renal form of trehalase was soluble and had a pI around 4.37. These pI values were consistent with those found in extracts of fetal intestinal (pI 4.60) and renal (pI 4.24) tissues. The determination of trehalase isozyme composition of amniotic fluid from pathological pregnancies with anal imperforation and polycystic kidney disease confirmed our findings on the origin of amniotic fluid trehalase. In the sample from a fetus with anal imperforation, low or absent intestinal trehalase isozyme was observed whereas a higher than normal level of renal trehalase activity was found in a fetus with polycystic kidney disease. PMID- 8222279 TI - Cystic fibrosis: the impact of analytical technology for genotype-phenotype studies. AB - The generalized exocrinopathy cystic fibrosis (CF) is the most common severe genetic disease in Caucasian populations. A panel of more than 700 chromosomes from German and Turkish CF patients was screened for disease-causing mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene by chemical cleavage of mismatch, single strand conformation polymorphism, restriction analysis and direct sequencing of genomic DNA amplified by polymerase chain reaction. Besides the major 3-bp deletion, delta F508 that was found on 73% of German CF chromosomes, more than 50 other missense, nonsense, frame-shift, and splice-site mutations have already been identified. In general, a CFTR mutation is linked with a single 10-marker haplotype which indicates that in most cases a particular mutation spread from a common ancestor. The comparison of mutation genotypes with the disease phenotype emphasized the causative role of the type and localization of the CFTR mutation for clinical course and prognosis. Pancreatic status and the risk of colonization of airways with opportunistic pathogens are genetically determined. Most patients who are harbouring mutations in the nucleotide binding folds were suffering from severe CF disease. Mild or even aberrant forms of CF were observed for many missense mutations located in the putative transmembrane domains or for mutations that are expected to result in a truncated protein of half of wild-type CFTR. PMID- 8222278 TI - Genotype-phenotype correlations in phenylketonuria. AB - Genotyping of the phenylalanine hydroxylating system offers a new way of characterizing patients with phenylalanine hydroxylase (PAH) deficiency. This paper investigates the power of genotyping as a parameter for differential diagnosis and as a measure of the risk factor of brain damage in well-treated patients with phenylketonuria (PKU). Thirty-three PKU patients were followed up over 9 years and the quality of dietary treatment, plasma phenylalanine (phe) in the newborn period before treatment and intellectual outcome at the age of 9 years were measured and correlated with the predicted residual activity (PRA) of the phe hydroxylase system as estimated from mutation analysis of the PAH gene. Patients were grouped in group Ia (PRA = 0%), group Ib (PRA = 5-15%) and group II (PRA > or = 25% of the normal activity). Mean plasma phe levels in the newborn in group Ia were 37.9 +/- 6.5 (2296 +/- 394), in group Ib 40.8 +/- 15.9 (2472 +/- 963) and in group II 16.2 +/- 4.2 (981 +/- 254) mg/dl (mumol/l). Difference in mean plasma values of groups Ia and Ib on the one hand and group II on the other were highly significant (P < 0.0001). No difference could be seen between groups Ia and Ib. There was a higher mean IQ at the age of 9 years in group II (97.4 +/- 5.4) in comparison with groups Ia (92.7 +/- 12.8) and Ib (85.0 +/- 14.4). The difference between group Ib and group II was significant (P < 0.040).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222280 TI - Variable phenotypic expression of genotypic abnormalities in the porphyrias. AB - The inherited porphyrias are the consequence of inherited deficiencies of enzymes in the heme synthesis pathway; they exhibit classical Mendelian inheritance patterns. The acute porphyrias (acute intermittent, porphyria variegata, hereditary coproporphyria) result from 50% (approx.) deficiencies of specific enzymes, which demonstrate autosomal dominant inheritance. However, only approx. 10% of subjects who inherit a porphyrin enzyme deficiency develop the corresponding acute porphyria and in most instances there is no obvious reason why one patient with an enzyme deficiency is symptomatic whereas another is not. Control of heme synthesis is achieved by the repressor effect of heme on the enzyme ALA synthase. Acute attacks of porphyria can be precipitated in susceptible persons by drugs, ethanol, starvation, hormones, stress and infection. The mechanism is usually by induction of ALA synthase activity. The molecular biology of porphyria variegata and hereditary coproporphyria is large unexplored. Acute intermittent porphyria is due to a partial deficiency of the enzyme porphobilinogen deaminase in the liver. The location of the gene for this enzyme has been identified on the long arm of chromosome 11. Acute intermittent porphyria is a genetically heterogenous disease with the abnormality frequently being a point mutation affecting synthesis of the enzyme. PMID- 8222281 TI - The diagnosis of inborn errors of metabolism by examination of the genotype. PMID- 8222282 TI - Comparative clinical evaluation of biochemical and genomic tumor markers. AB - The clinical utilities of established biochemical tumor markers and of emerging genomic markers are compared by six formal criteria: [1] tests negative in health or benign disease, [2] produced exclusively by specific tumor cells, [3] present frequently in the targeted malignancy, [4] detectable in occult disease, [5] degree of expression reflects tumor burden and prognosis, and [6] degree of expression correlates with therapeutic result. Evaluation of eight widely accepted marker systems combining a biochemical indicator with a specific cancer, on the one hand, and five representative genomic marker-target combinations involving chromosomal translocation, gene amplification and mutation, on the other hand, produces three main conclusions: First, specified applications are sufficiently well documented for the best biochemical markers to now tailor analytical performance goals to these uses. Second, further clinical trials of genomic markers are needed to document the useful linkage of specific indicators with specific clinical problems. Third, the different profiles of marker characteristics defining the two classes of indicators suggest some mutually complementary uses. PMID- 8222283 TI - Myc amplification: regulation of myc function. AB - The myc oncogenes have been implicated in the control of cell proliferation in both normal and neoplastic cells. There is increasing evidence that Myc proteins function as transcriptional regulators of other genes apparently involved in the control of cell proliferation. The effects of Myc on both gene expression and cell growth are differentially regulated by the recently described Max and delta Max proteins that can either cooperate or compete with Myc for sequence-specific DNA binding. PMID- 8222284 TI - Genetic and molecular heterogeneity of breast cancer cells. AB - We have undertaken a systematic study of primary human breast tumor DNAs to identify and characterize frequently occurring somatic mutations. Loss of heterozygosity (LOH) was found on chromosomes 1p (37%), 1q (20%), 3p (30%), 7 (41%), 13q (30%), 17p (49%), 17q (29%) and 18q (34%) in our tumor DNA panel. Specific subsets of tumors could be defined based on the particular collection of mutations they contained. One goal of these studies has been to determine whether there is a significant association between specific mutations and clinical parameters of the disease. We have found that LOH on chromosome 17p in tumor DNAs is associated with breast tumors having a high proliferative index and that LOH on chromosome 7 is associated with patients having a poor prognosis. Our analysis of chromosome 17 suggests that there may be as many as four tumor suppressor genes affected in primary human breast tumors. PMID- 8222285 TI - Detection of minimal residual leukemia by the polymerase chain reaction: potential implications for therapy. PMID- 8222286 TI - Molecular diagnostics: issues of utilization, regulation and organization. PMID- 8222287 TI - Endothelins as regulators of growth and function in endocrine tissues. PMID- 8222288 TI - Thyrotrophin receptor expression: does it help in assessing the prognosis of thyroid cancer? PMID- 8222289 TI - Expression of thyrotrophin receptor gene in thyroid carcinoma is associated with a good prognosis. AB - OBJECTIVE: The clinical course of thyroid carcinoma is very variable. It is well known that thyroid carcinomas of similar histology can behave differently in terms of local invasion and distant metastases: there is no reliable way to predict the disease course with confidence. In the present study we compared the TSH receptor and c-myc mRNA levels in different stages of thyroid carcinomas to identify whether they are useful markers for thyroid tumour biological behaviour and prognosis. DESIGN: Thyroid tumour specimens were used as the source of RNA. The TSH receptor and c-myc mRNA levels were detected by Northern blot analysis and quantitated by laser densitometry. PATIENTS: Thyroid tissues were obtained from five patients with multinodular goitres, 22 with differentiated and three with anaplastic carcinomas. MEASUREMENTS: Total cellular RNA was extracted from thyroid tissue specimens and blotted onto nylon membranes. Northern blot analysis was used to detect TSH receptor and c-myc mRNA. The mRNA levels were then quantitated by laser densitometry and compared with each disease stage. RESULTS: TSH receptor mRNA levels were significantly lower in carcinomas as compared to benign tumours. With advancing disease stage, the neoplastic tissues generally showed a progressive decline in TSH receptor mRNA levels. Interestingly, in two specimens from patients with distant metastases, TSH receptor mRNA levels were not significantly reduced and were comparable to those in benign tumours. Both patients are still alive, one of them 18 years after operation, indicating that tumour histology is dissociated from its biological behaviour. c-myc mRNA levels were increased but not significantly so in stage 1-3 carcinomas. However, in stage 4 carcinomas c-myc expression was significantly increased. Thus c-myc overexpression is associated with poor prognosis. Although there is a negative correlation between TSH receptor and c-myc mRNA levels, the correlation was not significant. CONCLUSIONS: These results indicate that decreased TSH receptor and increased c-myc gene expression levels are associated with thyroid cell de differentiation. They are useful markers for thyroid tumour de-differentiation and disease prognosis. PMID- 8222290 TI - The importance of thyroid microsomal antibodies in the development of elevated serum TSH in middle-aged women: associations with serum lipids. AB - OBJECTIVE: We assessed the relative risk of both serum TSH and antithyroid antibody concentrations with regard to progression of thyroid failure and studied the lipid profiles of individuals with elevated TSH levels. DESIGN, PATIENTS AND MEASUREMENTS: In a randomly selected group of 427 women aged 40-60 (mean 55) years volunteering in an epidemiological survey in Zoetermeer, TSH and thyroid microsomal antibodies (TMA) were determined. Ten years after the initial survey only TSH was measured and the lipid profiles of the individuals with elevated serum TSH levels were studied and compared with a reference group. RESULTS: During follow-up, four of 427 women were treated with thyroxine. Seventeen of 423 women initially had elevated serum concentrations of TSH (> 4.2 mU/l), 11 of whom were TMA positive. In the group of 406 women with initially a normal TSH, nine out of 37 (24%) TMA-positive women developed elevated serum levels of TSH over 10 years. In contrast only 10 of 369 (3%) TMA-negative women had elevated serum TSH levels 10 years after the initial survey (P < 0.001). Altogether, at the end of the observation period, 40% of TMA-positive subjects had elevated serum TSH concentrations, compared to 3% in the TMA-negative group (P < 0.01). TSH levels in the upper part of the normal range also appeared to have a predictive value: if those both with TSH levels between 2.0 and 4.2 and with a positive TMA status were contrasted with those without antibodies and low TSH, the crude relative risk was 71.5 (31.0-164.3), whereas the crude relative risk of presence versus absence of TMA was only 36.3 (18.8-70.3). Women with elevated TSH levels did not show changes in serum concentrations of total cholesterol (7.4 +/- 1.1 mmol/l), apo-A, (4.7 +/- 1.0 mmol/l) and apo-B (3.1 +/- 0.7 mmol/l) lipoproteins, compared with control individuals (7.2 +/- 1.3, 4.7 +/- 0.8 and 3.0 +/- 0.7 mmol/l, respectively). CONCLUSIONS: The determination of serum TMA in middle-aged women can identify an important group of women at risk of developing an elevated serum TSH. TMA measurement is of potential use in the prevention of cardiovascular disease. An elevated serum TSH, however, in our study-group does not seem to be accompanied by an abnormal lipid profile, as reported by others. PMID- 8222291 TI - Prospective study of the hypothalamic-pituitary axis in thalassaemic patients who developed secondary amenorrhoea. AB - OBJECTIVE: We aimed to prospectively evaluate during 10 years the GnRH gonadotrophin secretory dynamics in a cohort of 15 menstruating girls with beta thalassaemia major to determine whether they sustained progressive damage to this axis. DESIGN: Patients were characterized by 12-hour gonadotrophin profiles (by sampling blood at 15-minute intervals) and assessment of gonadotrophin responses to 100 micrograms GnRH bolus (by sampling blood at 20-minute intervals for 1 hour and at 2 hours) sequentially during the follicular and luteal phases of their menstrual cycles, 12-14 months and 5-6 years after the onset of secondary amenorrhoea. SUBJECTS: We studied 15 post-menarcheal thalassaemic girls and five age-matched control subjects who were the healthy siblings of the patients. MEASUREMENTS: FSH and LH assays were determined using commercial RIA systems and double antibody techniques. Pulse detection used the Pulsar technique of Merriam and Wachter. RESULTS: We demonstrated that during their amenorrhoeic period, all thalassaemic patients had gonadotrophin pulse abnormalities and low-normal GnRH stimulated gonadotrophin levels indicating that they had GnRH-gonadotrophin secretory insufficiency. During the subsequent 10 years there was progressive deterioration of hypothalamic-pituitary function in all patients; 66% became apulsatile and all had marked reduction in their GnRH-stimulated gonadotrophin levels. CONCLUSIONS: Our investigation suggests that thalassaemic patients with secondary amenorrhoea had severe and progressive damage to their hypothalamic pituitary axes despite intensive chelation therapy. PMID- 8222292 TI - Adrenocorticotrophin and cortisol secretion in children after low dose cranial irradiation. AB - OBJECTIVE: We investigated the effect of low dose cranial irradiation (18-24 Gy) on spontaneous ACTH and cortisol secretion in children. DESIGN: We analysed 24 hour plasma ACTH and cortisol profiles sampled at 20-minute intervals. PATIENTS: Twenty long-term survivors of acute lymphoblastic leukaemia were studied and results compared with those in 14 normal children. MEASUREMENTS: ACTH and cortisol profiles were analysed by Fourier transformation and spectral analysis of stationarized data, autocorrelation and coherency analysis. RESULTS: The normal circadian rhythms of ACTH and cortisol were preserved in the children after cranial irradiation. The median 0900 h and midnight values were 1.50(0.8 6.4)pmol/l and 1.0(0.6-3.7)pmol/l respectively for ACTH and 282(48-1913)nmol/l and 57.5(44-637)nmol/l respectively for cortisol, and were not significantly different from those in the normal group. Fourier transformation revealed dominant periodicities for ACTH at 0.7-1.1 h, equivalent to 22-34 ACTH secretory bursts per 24 hours, and for cortisol at 0.7-1.1 h and 2-4.8 h. Similar results were found in the normal group. Coherency analysis indicated a significant shared periodicity of 0.7-1.2 h in nine children, corresponding to 20-34 related secretory bursts in 24 hours for ACTH and cortisol. After pooling the coherency spectra in the cranially irradiated group, comparison with the pooled data from the normal group revealed no significant difference between the two groups in the relationship between the two hormones. CONCLUSION: No significant disruption of spontaneous ACTH or cortisol secretion, either in the amount or pattern of hormones secreted, was found in children after low dose cranial irradiation (18 24 Gy). PMID- 8222293 TI - Comparison of computerized tomography and magnetic resonance imaging for the examination of the pituitary gland in patients with Cushing's disease. AB - OBJECTIVE: The vast majority of patients with Cushing's disease have a corticotroph adenoma, the selective removal of which, through the transsphenoidal route, has the potential to offer a definitive and complete cure. This study was designed to compare the diagnostic accuracy of computerized tomography (CT) and magnetic resonance imaging (MRI) to identify the presence, evaluate the size, and assess the topographic characteristics of pituitary corticotroph adenomas. METHODS: Forty-two patients with Cushing's disease were included in this prospective study, of whom 16 were subsequently explored transsphenoidally. Computerized tomography used a CE 12,000 CGR apparatus with 1.5-mm coronal slices and a 2-mm interslice gap. Magnetic resonance imaging used an MR Max 0.5T GE apparatus; adjacent 3-mm slices were obtained with a T1-weighted gradient echo before and after gadolinium injection. RESULTS: Lesions compatible with an adenoma were identified in 29 patients by MRI and in 21 patients by CT (69 vs 50%, P < 0.02). Seven macroadenomas were identified as well by the two methods. Eight of the 22 microadenomas detected by MRI were not identified by CT. Evidence for intracavernous tumour extension was found in nine patients: it was more frequently detected by MRI (8 patients) than by CT (4 patients). Fourteen patients with positive MRI had a pituitary examination: in one case the adenoma could not be reached because of purely suprasellar location; of the other 13 all were found by the surgeon and the surgical outcome was successful in 12 cases. CONCLUSION: MRI is superior to CT for the examination of the pituitary gland in patients with Cushing's disease. PMID- 8222294 TI - Bilateral adrenalectomy: low mortality and morbidity in Cushing's disease. AB - OBJECTIVE: We assessed the current role of bilateral adrenalectomy in the overall management strategy of hypercortisolism. DESIGN: Retrospective review of case notes. PATIENTS: Twenty-six patients (20F/6M); mean age 46 years (range 15-70 years), median duration of follow-up 5.25 years (0.6-19.1 years) who had undergone bilateral adrenalectomy at the Royal Victoria Hospital since 1972. Eighteen had had prior transsphenoidal surgery which did not control the hypercortisolism. MEASUREMENTS: Morbidity, mortality, incidence of Nelson's syndrome. RESULTS: Surgery was performed through bilateral postero-lateral incisions (20 patients) or a long epigastric incision (six patients). The mean combined weight of the adrenals at surgery was 11.2 g. Twenty patients received subcutaneous heparin and 18 antibiotic prophylaxis peri and post-operatively. There was no operative mortality. Minor complications included one post-operative wound infection and a small pneumothorax requiring drainage. Major complications occurred in two other patients, both with pre-existing invasive pituitary tumours and considered at high risk because of age and general debility. One patient had a massive pulmonary embolus and the other a subphrenic abscess post-operatively. This latter patient, the only mortality, died from an unrelated cause three years post-operatively. Six patients have subsequently undergone pituitary surgery and three have received external pituitary irradiation therapy for expanding tumours. CONCLUSIONS: Bilateral adrenalectomy, in experienced hands, is a relatively safe and useful management option in patients with hypercortisolism. Growth of a pituitary adenoma post-operatively is now the most worrying complication. PMID- 8222295 TI - The short and long-term effects of octreotide on calcium homeostasis in patients with acromegaly. AB - OBJECTIVE: The somatostatin analogue octreotide (Sandostatin, Sandoz) is effective in reducing growth hormone levels in patients with acromegaly. Early and transient gastrointestinal side-effects are frequent. The aim was to evaluate whether gastrointestinal side-effects during the initial phase of octreotide treatment affect calcium homeostasis, and whether effects on calcium homeostasis are seen during long-term treatment with octreotide in patients with optimal effect on GH and subjectively normal gastrointestinal function. DESIGN: We first studied short-term treatment with octreotide during 14 days. From day 15 of the study medication was withdrawn, and on day 20 follow-up measurements were made. We then observed the effects of long-term treatment with octreotide. Mean duration of treatment until the day of blood sampling was 32 months (range 9-48 months). PATIENTS: Sixteen patients with acromegaly were studied, ten in the short-term study and ten in the long-term study; four were included in both. MEASUREMENTS: Serum levels of calcium, phosphate, PTH, alkaline phosphatase, 1,25(OH)2 Vit D, vitamin D-binding protein and sex hormone-binding globulin (SHBG) were measured before treatment (day 0) and on days 4, 6, 8, 14 and 20 during the short-term study and, except SHBG, before treatment and during therapy in the long-term study. RESULTS: During the short-term treatment mean (+/- SEM) serum calcium decreased significantly (on days 6 and 8, 2.21 +/- 0.08 and 2.15 +/ 0.09 mmol/l, respectively vs basal level, 2.38 +/- 0.08 mmol/l), whereas significant increments were seen in mean serum PTH (on day 14, 36 +/- 4 vs basal, 24 +/- 3 ng/l; ng/l divided by 9.425 = pmol/l), mean serum 1,25(OH)2 Vit D (on day 8, 112 +/- 7 vs basal 96 +/- 8 pmol/l), and 'free 1,25(OH)2 Vit D-index', i.e. molar ratio of 1,25(OH)2 Vit D and vitamin D-binding protein (on days 8 and 14, 1.72 +/- 0.09 x 10(-5) and 1.66 +/- 0.11 x 10(-5), respectively vs basal, 1.33 +/- 0.09 x 10(-5)). The changes were within the normal range. No changes were seen in serum phosphate, alkaline phosphatase, or vitamin D-binding protein. During the long-term study mean serum calcium and phosphate decreased significantly, 2.32 +/- 0.04 vs basal 2.42 +/- 0.04 mmol/l and 1.24 +/- 0.07 vs basal 1.40 +/- 0.09 mmol/l, respectively, whereas mean serum PTH increased significantly, 40 +/- 8 vs basal 21 +/- 5 ng/l. The changes were within the normal range. No changes were seen in serum alkaline phosphatase, 1,25(OH)2 Vit D, free 1,25(OH)2 Vit D-index, or vitamin D-binding protein. CONCLUSION: Altered calcium homeostasis during octreotide treatment in acromegaly is not only initial and temporary, but can also be seen after several years of treatment. The clinical relevance of these long-standing effects needs to be further investigated. PMID- 8222296 TI - Metabolic clearance rate of biosynthetic growth hormone after endogenous growth hormone suppression with a somatostatin analogue in chronic renal failure patients and control subjects. AB - OBJECTIVE: Several disturbances in the regulation of growth hormone secretion have been reported in chronic renal failure. The general assumption is that an altered hormonal clearance is at the basis of such GH alterations. Nevertheless, details of GH elimination kinetics in uraemia are not available. To clarify the role played by the kidney in its catabolism, GH elimination kinetics were studied in uraemic and control subjects after suppression of endogenous secretion of GH. DESIGN: In all subjects an analogue of somatostatin (octreotide 100 micrograms i.v.) was administered as a bolus before GH (-60 minutes). Sixty minutes later (0 min) biosynthetic GH (0.5 IU = 200 micrograms) was administered intravenously as a bolus. PATIENTS: Six chronic renal failure patients before dialysis and six matched normal volunteers. MEASUREMENTS: Plasma GH levels were measured by an immunoradiometric assay. RESULTS: In both groups, the GH elimination curve fitted a bi-exponential model. The calculated plasma volume and GH concentration at 0 minutes were similar in both groups, while uraemic patients presented a reduced distribution volume. In all parameters measuring GH elimination, chronic renal failure patients showed an impaired clearance. In fact, the area under the curve (mU/l/150 min) was 912.8 +/- 170.6 for controls and 3524.8 +/- 642.8 for chronic renal failure patients (P < 0.005). The GH half-life was 13.8 +/- 1.6 and 26.4 +/ 2.9 minutes for control and uraemic subjects respectively (P < 0.05), and the metabolic clearance rate MCR (ml/min/m2) was 265.3 +/- 50.6 for controls and 79.9 +/- 16.4 for uraemic patients (P < 0.05). The GH mean residence time (minutes) (MRT) calculated was 12.0 +/- 0.5 for controls and 31.8 +/- 4.6 for chronic renal failure patients (P < 0.05). CONCLUSIONS: Contrary to previous estimates, GH elimination kinetics follows a bi-exponential model and in normal subjects the GH half-life of the second phase is 13.8 +/- 1.6 minutes. Uraemic patients have impaired clearance of GH, suggesting that the kidney plays a role in GH disposal. However, the degree of impairment does not fully explain the alterations in GH secretion previously described in chronic renal failure. PMID- 8222297 TI - Hyperactivity of the hypothalamo-pituitary-adrenal axis in obesity: a study of ACTH, AVP, beta-lipotrophin and cortisol responses to insulin-induced hypoglycaemia. AB - OBJECTIVE: The purpose of this study was to determine whether alterations in the hypothalamo-pituitary-adrenal axis and arginine vasopressin secretion, which have been associated with animal obesity, also occur in man. DESIGN: Cross-sectional analysis of extremely obese women and normal weight controls. PATIENTS: Thirty three obese premenopausal, non-diabetic women (mean age 31 years, mean body mass index (BMI) 41), and 15 normal weight controls (mean age 24 years, mean BMI 22). MEASUREMENTS AND RESULTS: Arginine vasopressin (AVP), ACTH, beta-lipotrophin and cortisol responses to insulin-induced hypoglycaemia (0.2 units Actrapid/kg body weight for obese; 0.15 unit/kg for controls) were measured. The obese women were further characterized by anthropometric measurements (weight, body mass index, fat distribution) and indices of insulin secretion/resistance: fasting insulin, insulin secretion during 75-g oral glucose tolerance test area under curve, insulin-stimulated glucose disposal and an index of insulin resistance. No significant differences were found in the basal levels of ACTH, AVP, beta lipotrophin or cortisol. An augmented peak beta-LPH (n = 16, P < 0.02, the difference of the mean 3.65, 95% confidence interval 1.33-10) and ACTH (n = 16, P = 0.05, the difference of the mean 2.12, 95% CI 1.0-4.5) response were found in obese as compared with normal weight controls. Both ACTH and AVP areas under the curve were similar in both groups studied. There was additionally a direct positive association between the integrated ACTH response (area under the curve) and the weight of the obese subjects (P < 0.05, r2 = 0.265). The cortisol response was negatively correlated with insulin-stimulated glucose disposal (P < 0.01, r2 = 0.23), but not with other indices of insulin secretion/resistance (fasting insulin, oral glucose tolerance test area under the curve, index of insulin resistance) or fat distribution. Comparable responses to hypoglycaemia were seen for AVP and cortisol. There was no correlation between the ACTH, AVP or cortisol responses. CONCLUSION: Obesity is associated with increased activity of the hypothalamo-pituitary-adrenal axis as supported by augmented ACTH and beta lipotrophin secretion in response to insulin-induced hypoglycaemia and the positive association between the ACTH response and the body weight of obese women studied. PMID- 8222298 TI - The relationship of insulin insensitivity to menstrual pattern in women with hyperandrogenism and polycystic ovaries. AB - OBJECTIVE: Insulin insensitivity is a recognized feature of polycystic ovary syndrome (PCOS) but previous studies have suggested that circulating insulin concentrations are normal in hyperandrogenaemic women with regular cycles. The aim of this study was to examine the relationship between insulin sensitivity and menstrual pattern in women with PCO. DESIGN: A cross-sectional study of insulin sensitivity in a cohort of PCO subjects with oligomenorrhoea compared to women with PCO and regular menstrual cycles and a group of normal control subjects. SUBJECTS: Seventy-two women with polycystic ovaries on ultrasonography were studied. PCO subjects had clinical and/or biochemical evidence of hyperandrogenism; 53 had oligo/amenorrhoea (olig) and 19 had regular menses (reg). Results were compared with 31 control subjects. The groups were matched for age, weight and ethnic origin. METHODS: Glucose and insulin responses to 75 g oral glucose were measured. Insulin sensitivity was assessed by the decline in plasma glucose following intravenous insulin (0.05 U/kg). RESULTS: Glucose area (mean +/- SEM) after oral glucose was increased slightly in both PCO groups compared with controls (olig 37.6 +/- 1.4, reg 36.0 +/- 1.8, control 33.7 +/- 0.9 mmol/l h, both P < 0.01). Insulin area median (interquartile range) in response to glucose was significantly greater in the oligomenorrhoeic group (346 (239-734) mU/l h), compared with both PCO with regular cycles (246 (148-355), P < 0.01) and controls (221 (147-277), P < 0.01). Insulin sensitivity was reduced (P < 0.01) in the oligomenorrhoeic group (147 +/- 9.2 mumol/l min) compared to controls (185 +/ 7.4) but was normal in PCO with regular cycles (182 +/- 12.5). Insulin sensitivity did not correlate significantly with plasma testosterone or with SHBG levels, but plasma insulin concentrations correlated negatively with SHBG levels (fasting insulin vs SHBG, r = -0.47, P < 0.01; insulin area vs SHBG, r = -0.41, P < 0.01). CONCLUSIONS: Insulin insensitivity in polycystic ovary syndrome occurs when there is oligo/amenorrhoea but not when the menstrual cycle is regular. This is consistent with PCO and insulin insensitivity being separate abnormalities which when combined are associated with anovulation. PMID- 8222299 TI - Pharmacokinetic properties of the tocolytic agent [Mpa1, D-Tyr(Et)2, Thr4, Orn8] oxytocin (antocin) in healthy volunteers. AB - OBJECTIVE: The aim of this study was to study the pharmacokinetics of antocin, the tocolytic oxytocin antagonist [Mpa1, D-Tyr2(Et), Thr4, Orn8]-oxytocin. DESIGN: Antocin was injected intravenously as a bolus dose (5 mumol). Blood samples were taken at intervals for 240 minutes. In addition, the binding of 125I Tyr10-antocin to blood constituents was determined and compared with 125I-AVP and 125I-[Mpa1, D-Arg8]-vasopressin (desmopressin). SUBJECTS: Eight healthy, non smoking adults, three male and five female. MEASUREMENTS: Antocin was measured using a specific radioimmunoassay after prior extraction of the plasma. Plasma binding was estimated using polyethyleneglycol precipitation. RESULTS: The rate of plasma disappearance of antocin was best fitted by a biexponential curve. The clearance of antocin was 23.5 +/- 7.6 l/h, the volume of distribution was 13.1 +/ 3.8 l and the biological half-life was 39.0 +/- 4.1 minutes. A greater proportion of 125I-Tyr10-antocin bound to plasma proteins (33.5%) and red blood cells (13%) than did 125I-AVP, 125I-desmopressin and unlabelled desmopressin. CONCLUSIONS: The half-life was longer and the clearance of antocin was less than that found in a previous study when a non-specific antiserum was used. This is most likely because of the extended blood sampling time period which revealed the biphasic decay pattern. The higher plasma clearance of antocin compared to oxytocin and desmopressin may be explained by its increased binding to blood constituents rather than by differences in enzymatic degradation of the molecules. PMID- 8222300 TI - ACTH-independent Cushing's syndrome in pregnancy with spontaneous resolution after delivery: control of the hypercortisolism with metyrapone. AB - A 25-year-old primigravid woman presented with Cushing's syndrome at 23 weeks gestation; serum cortisol was 1090 nmol/l at 0900 h, 1230 nmol/l at 2200 h; basal urinary free cortisol excretion was 3680 nmol/24 h, and 8830 nmol/24 h after dexamethasone 8 mg daily for 48 hours; plasma ACTH was < 1.1 pmol/l. CT scan of the adrenal glands showed bilateral adrenal hyperplasia. The hypercortisolism was controlled with metyrapone until elective delivery of the fetus by Caesarean section at 34 weeks gestation because of a decline in growth. No adverse fetal effects of metyrapone treatment were apparent, maternal outcome was uncomplicated and wound healing was unimpaired. Maternal adrenocortical function had returned to normal within 4 weeks of the cessation of pregnancy and biochemical remission has been maintained up to 9 months post-partum. Metyrapone therapy is effective in controlling the hypercortisolism in certain cases of Cushing's syndrome complicating pregnancy. PMID- 8222301 TI - Isolated congenital ACTH deficiency: a cleavage enzyme defect? AB - The pro-opiomelanocortin (POMC) gene encodes adrenocorticotrophin (ACTH) which is derived from precursors by proteolytic cleavage. Congenital, isolated ACTH deficiency is rare but may be familial and fatal. The aetiology is unknown though defects at both hypothalamus and adenohypophysis have been postulated. We have studied a female presenting with hypoglycaemia in the neonatal period. When studied at 6 weeks of age, ACTH was unmeasurable even after injection of corticotrophin releasing hormone (CRH1-41). ACTH precursors, quantitated by two site immunoradiometric assay, were clearly measurable prior to treatment and were stimulated by CRH1-41 and suppressed by glucocorticoid administration. Concentrations of POMC, N-terminal pro-opiocortin (N-POC) and beta-endorphin (beta-EP) were within the normal adult range during glucocorticoid replacement therapy; ACTH and beta-lipotrophin remained undetectable. The secretion of glucagon, measured by radioimmunoassay, in response to hypoglycaemia was normal. By sequencing polymerase chain reaction products from the patient's genomic DNA, the entire coding region of the POMC gene was established to be normal. The results are compatible with a cleavage enzyme defect. PMID- 8222302 TI - Isolated ACTH deficiency: enzyme defect or chimaeric enzyme? PMID- 8222303 TI - The surgical treatment of megaesophagus and megacolon. AB - The effect of Chagas' disease on the digestive tract is evaluated based on the experience acquired in the surgical management of 929 cases of megaesophagus: 807 in the nonadvanced state of the disease and 122 with dolichomegaesophagus. The 807 subjects with nonadvanced megaesophagus were submitted to wide esophagocardiomyectomy on the anterior esophagogastric junction combined with an antireflux valvuloplasty procedure. There was no mortality. On the other hand, dolichomegaesophagus was always resected, with an esophagogastroplasty through the esophageal bed. The index of mortality of the series was 4.1% (5/122) and the main postoperative complications were pleural effusion (22.1%) and fistula of the esophagogastric anastomosis (8.2%). Over the long term the patients adapted well to the operation and gained weight. For the treatment of the megacolon, an easier technique has been employed: resection of the dilated sigmoid colon and colorectal anastomosis just above the anorectal ring. In 30 patients, no leakages, infection or other major complications were seen. The patients, after 1 year of follow-up, had a normal bowel transit. PMID- 8222304 TI - Endoscopic hemostasis of bleeding peptic ulcers. AB - Developments in therapeutic endoscopy over the last decade have made it possible to perform endoscopic hemostasis for bleeding peptic ulcers. This review traces the developments in this field in the Department of Surgery of the National University of Singapore. A recently conducted prospective randomized controlled trial using intralesional adrenaline and heater probe demonstrated that initial hemostasis could be achieved in 100% of patients with actively bleeding ulcers or stigmata of recent hemorrhage. The rebleed rate was 6.6% compared with 20.3% in a well-matched control group. The recent advent of laparoscopic vagotomy and laparoscopic Billroth II gastrectomy offers a low-morbidity surgical option to long-term medical treatment for the follow-up management of patients with bleeding ulcers. PMID- 8222305 TI - Anal cancer--current perspectives. AB - Anal cancer is uncommon accounting for only 2% of anorectal cancers. The recognition of many similarities between cervical and anal cancer has stimulated research into the identification of a common aetiological agent. DNA from human papillomaviruses has consistently been found in both of these cancers and is thought to be an important factor in the development of both of these tumours. Simultaneously, epidemiological data from the west coast of America have indicated that the demography of anal cancer may be changing. Further studies in the USA and the UK have identified certain groups at high risk of developing anal cancer. These high-risk groups include 'never married' men and immunosuppressed patients both from iatrogenic immunosuppression in transplant patients and those infected with HIV. The potential increase in anal cancer cases, due to the ever increasing numbers of patients who have received transplants and the spiralling number of the population infected with HIV make it timely to review what is known of the aetiology, presentation and management of this cancer. PMID- 8222306 TI - Anorectal manometry: the state of the art. AB - This review deals first with the methodology of anorectal manometry. The methods of recording anorectal pressures with perfused catheters, sleeve catheters, water or air-filled balloon catheters and microtransducers are presented and discussed as well as the various polygraphs used in this investigation. In the second part, the routine anorectal manometry and the parameters to be studied in this investigation are described. Then, the additional manoeuvres which have been described in anorectal manometry are discussed. Finally, the major results obtained in recto-anal dysfunctions are described, that is manometric findings in constipation in adults and children, in incontinence and in the descending perineum syndrome. The usefulness of anorectal manometry in surgical and various conditions is also discussed. PMID- 8222307 TI - Acetaldehyde/protein interactions: are they involved in the pathogenesis of alcoholic liver disease? AB - Alcohol abuse is a major cause of liver disease. While ethanol itself has been shown to be hepatotoxic, its primary metabolite acetaldehyde has also been implicated in the pathogenesis of alcoholic liver disease. The majority of ethanol metabolism occurs in the liver and high concentrations of acetaldehyde accumulate during chronic ethanol abuse. Acetaldehyde has been shown to react with many proteins in vitro, forming stable covalent adducts. These modifications can act as neoantigens and may also alter biological function. Acetaldehyde modified proteins have been detected in the livers of ethanol-fed rats and human alcoholics. Circulating antibodies reactive with modified proteins have also been detected. A direct linkage between acetaldehyde-modified proteins, antibodies and liver damage has yet to be established, but current research should clarify the picture in the next few years. PMID- 8222308 TI - Biliary surgery without the surgeon! AB - Since the introduction of endoscopic retrograde cholangiopancreatography in the 1970s and of endoscopic sphincterotomy (EST) in 1974, endoscopic techniques for the diagnosis and therapy of biliary and pancreatic disorders have proliferated. Although some procedures have become part of routine practice, for example EST for postcholecystectomy bile duct stones and stent insertion for inoperable biliary strictures, the place of others is unclear at present and should be evaluated by prospective randomized clinical trials. The choice of either an endoscopic or a transhepatic approach for biliary disease is usually dictated by local expertise, but these should be regarded as complementary rather than competing techniques, and complex biliary problems should be managed jointly by the endoscopist, interventional radiologist and hepatobiliary surgeon. PMID- 8222309 TI - Diarrhoea-causing Escherichia coli. AB - Current knowledge of Escherichia coli strains responsible for bowel infections, including the more recently defined entero- hemorrhagic and entero-adherent types, is reviewed. PMID- 8222310 TI - The intestinal immune system and its relation to disease. AB - The essential protective structure against the heavy enteric antigenic burden, the gut mucosa, prevents penetration of noxious agents, but allows a minimal exchange of large molecules and particles between the gut lumen and the 'milieu interieur' of the body. M cells in the follicle-associated epithelium of the gut, ideal gateways for the presentation of enteric antigens to the cells of the gut associated lymphoid tissue (GALT), are also weak links in the mucosal barrier, and may provide access for various microorganisms. The afferent limb of the GALT consists of distinct aggregates of lymphoid cells located in Peyer's patches, the vermiform appendix and the solitary lymphatic follicles, and of the mesenteric lymph nodes. The efferent limb subsumes the diffusely scattered mucosal leukocytes, mainly lymphocytes and plasma cells. Intraepithelial and mucosal T lymphocytes are instrumental in launching local immune responses, producing lymphokines, and in the specific lysis of virally infected cells. Antigenic stimulation of the GALT results in local secretion of antibodies, or in suppression of systemic immunologic responses to ingested antigens ('oral tolerance'). Poorly controlled mucosal immune responses result in organ-specific diseases. Extranodal lymphomas that mimic structures of the GALT may arise on a background of inflammatory or immunologic (autoimmune) disorders. PMID- 8222311 TI - Gastro-intestinal crises during pregnancy. AB - In general pregnancy is not associated with an increase in the incidence of gastro-intestinal (GI) conditions, but it is associated with an increase in the severity of these conditions. This is largely due to a delay in making the diagnosis. Delay is caused by a combination of factors. These include mimicry of symptoms and signs with pregnancy-related conditions, a change in the usual clinical presentation, reluctance to use radiological diagnostic aids and a higher threshold for performing invasive investigative procedures. However before resorting to invasive investigations, it is imperative that appropriate non invasive investigations are performed and evaluated first. This is especially relevant to liver conditions in pregnancy which can present with abdominal pain. Optimal management of GI conditions in pregnancy requires both surgeons and obstetricians to be aware of the problems of diagnosis, communicate promptly and investigate without delay in order to make a diagnosis and avert a GI crisis. This article discusses the clinical presentation, diagnosis and management of the common GI conditions occurring in pregnancy. PMID- 8222312 TI - Rheumatoid factor production by mononuclear cells derived from different sites of patients with rheumatoid arthritis. AB - To investigate the origin of circulating rheumatoid factor (RF) and the relation between RF production at different sites in patients with rheumatoid arthritis (RA), mononuclear cells derived from bone marrow, synovium and peripheral blood of patients with RA were examined for the presence of plasma cells and for their capacity to produce RF and other immunoglobulins in vitro. Analysis of culture supernatants for the presence of immunoglobulins demonstrated that cells derived from bone marrow, synovium and peripheral blood were all found to be capable of producing every immunoglobulin and RF isotype investigated. No significant correlations were found between concentrations of immunoglobulin isotypes produced by cells derived from different sites of one individual. Significant correlations were found, however, between concentrations of RF isotypes produced by cells derived from the three sites. These results indicate that the production of RF in the different compartments is not an autonomously regulated process. Mononuclear cells derived from bone marrow were found to be able to produce RF in similar quantities to cells dissociated from synovial tissue. In combination with the fact that circulating immunoglobulins are produced mainly in the bone marrow, this observation suggests that bone marrow is also a major source of circulating RF. PMID- 8222313 TI - The suppressive effect of gelatin-conjugated superoxide dismutase on disease development and severity of collagen-induced arthritis in mice. AB - We studied the effect of superoxide dismutase (SOD) on murine collagen-induced arthritis (CIA), an animal model of human rheumatoid arthritis (RA). Among SOD derivatives studied, only gelatin-SOD conjugate which has prolonged half life in vivo was effective to suppress the development of CIA, while native SOD or gelatin carrier alone was ineffective. Interestingly, pyran polymer-conjugated SOD which also has a long half life showed no suppressive effect on the disease. No significant effect on immune response against type II collagen (CII) was found in any of the experimental groups. In addition, induction of suppressor cells was not detected in spleen or lymph node cells of the gelatin-SOD-treated group. Therefore, these results suggest that oxygen radicals may have an important role in the effector phase of the immune response to manifest this chronic autoimmune polyarthritis. Thus, the use of appropriate antioxidants for the treatment of human RA may be rationalized. PMID- 8222314 TI - Comparison between the protective effects of mycobacterial 65-kD heat shock protein and ovomucoid in pristane-induced arthritis: relationship with agalactosyl IgG. AB - The IgG of patients with rheumatoid arthritis and mice with pristane induced arthritis (PIA) tends to lack the terminal galactose normally on the conserved N acetylglucosamine linked beta 1-2 to mannose in IgG. The terminal N acetylglucosamine (GlcNAc) residues of oligosaccharides on agalactosyl IgG may be an important component of the action of these glycoforms. Here, administration of ovomucoid, a glycoprotein rich in terminal GlcNAc, before pristane injection was found to reduce the incidence of PIA. This observation is the second report of an intraperitoneally administered antigen that reduces the incidence of PIA, mycobacterial 65-kD heat shock protein (hsp65) being the first. The suppressive effect of ovomucoid was not transferred from protected to naive recipients by spleen cells at the dose tested. By contrast, transfer of spleen cells from hsp65 protected mice to naive recipients conferred protection and this protection may be antibody-mediated. It is considered that ovomucoid and hsp65 protect against the development of PIA by different mechanisms. PMID- 8222315 TI - Effect of pregnancy on proteoglycan-induced progressive polyarthritis in BALB/c mice: remission of disease activity. AB - Proteoglycan-induced arthritis is a murine autoimmune model displaying many similarities to human rheumatoid arthritis and ankylosing spondylitis, as has been documented by clinical, immunological and histopathological studies. Since the onset of arthritis correlates with the serum antibody level to mouse cartilage proteoglycan (PG), it is believed that these autoreactive antibodies may play crucial roles in the pathological mechanisms of PG-induced arthritis. We have found that fertility in these PG-induced arthritic mice had been reduced but, unlike collagen-induced arthritis, had not been completely lost. Moreover, pregnancy had a beneficial effect upon the clinical symptoms with very little or no influence on serum antibody levels. Although fertility was retained and arthritic mothers delivered healthy offspring, the birth frequency was significantly less than in non-arthritic age-matched controls. Furthermore, the presence of anti-PG autoantibodies (predominantly IgG1 subclass) transmitted from arthritic mothers to infants transplacentally and by milk during the lactation period did not render these offspring either resistant or more sensitive to subsequent induction of arthritis. Subsequent immunization of infants with 'arthritogenic' PG revealed an unaltered susceptibility to arthritis induction. PMID- 8222316 TI - Critical role of peripheral blood phagocytes and the involvement of complement in tumour necrosis factor enhancement of passive collagen-arthritis. AB - Studies have implicated tumour necrosis factor-alpha (TNF-alpha) in type-II collagen (CII)-induced arthritis (CIA), a well established animal model of human rheumatoid arthritis. Precisely how TNF is involved in CIA is not yet clear. In this study the effects of TNF on CIA were examined, independent of its potential effects on the immune response, by performing peri-articular injection of TNF in combination with passive immunization of rats. A sub-arthritic dose (5 mg) of affinity-purified anti-CII IgG, which alone was insufficient to induce spontaneous clinical arthritis, was used throughout the study. Obvious clinical arthritis that persisted for several days was rapidly induced by injections of 100 ng TNF into hindpaws of rats that were passively immunized shortly before the TNF injection. Injections of TNF in non-immunized control rats did not induce clinical arthritis, nor did buffer-only injections in passively immunized controls. The clinical arthritic response was a local phenomenon, limited only to the TNF-injected hindpaws. No swelling was observed in the opposite, buffer injected hindpaws, indicating the effects of TNF were not systemic. Depletion of peripheral blood phagocytes with anti-rat neutrophil antiserum before passive immunization completely abolished the ability of TNF to induce clinical arthritis, identifying phagocytic cells as the essential target cells in evoking this arthritic response. A role for complement activation was also demonstrated in this model through the use of a soluble recombinant version of CD35, the cell surface complement receptor type-1 (sCR1, BRL55730), which significantly reduced TNF-induced arthritis in phagocyte-replete rats. PMID- 8222317 TI - Synthesis of the third component of complement (C3) by human gastric cancer derived cell lines. AB - This is a study of complement components secreted by gastric cancer-derived cell lines (MKN28, MKN45, MKN74 and KATO-III), each of which has a different histological origin. Haemolytic activity of complement component was detected only in the culture supernatant of KATO-III (C2 activity) and in that of MKN45 (C5 activity). However, the third component of complement, C3, was detected by an ELISA assay in the supernatants of all cell lines. In our studies focusing on C3 production by these cell lines, we have found that: (i) tumour necrosis factor (TNF) induced an increase in the amount of secreted C3 in a dose- and time dependent fashion; (ii) TNF (10 U/ml) stimulated C3 secretion by these cell lines to levels of 25.4-62.9 ng C3/10(6) cells per 24 hours; (iii) C3 haemolytic activity was detected in supernatants of TNF-stimulated cell lines. The mean specific activities of C3 by TNF (10 U/ml)-stimulated cell lines were 1.2-5.6 x 10(5) effective molecules/ng (e.m./ng), when that of C3 in normal human serum (NHS) was 1.7 x 10(6) e.m./ng; (iv) de novo synthesis of C3 by these cell lines was demonstrated by the effect of cycloheximide and by the incorporation of 35S methionine into secreted C3; (v) immunoblot analysis of culture supernatants indicated that secreted C3 was mainly composed of C3 alpha and C3 beta chains, but pro-C3 was also present. These results, which show the de novo synthesis and secretion of C3 by all the tested gastric cancer-derived cell lines in response to TNF, suggest the possibility that C3 may be secreted in the gastric wall as part of its normal physiology, or as a result of tumour pathology, and thereby participate in local immune or inflammatory responses. PMID- 8222318 TI - Why do patients develop reactions to streptokinase? AB - Minor reactions to streptokinase are not uncommon, although the etiology is unknown. It is widely presumed, however, that these, like the more serious immune reactions, are antibody-mediated. We measured specific anti-streptokinase IgG, subclasses IgG1, IgG2, IgG3, IgG4 and IgE by ELISAs, haemagglutination, indirect Coombs' test and immunoblotting in six patients who developed reactions to streptokinase. Evidence of complement activation by streptokinase was sought by a haemolytic complement assay and by measurement of C3, C4 and C3d. The patients who reacted to streptokinase presented with low titres of anti-streptokinase IgG (median = 5; range 0-32) and IgG1 (median = 3; range 0-14). No evidence of any other IgG subclass was found, nor of specific anti-streptokinase IgE. Anti streptokinase IgG1 was found to fix complement; patients who reacted to streptokinase were found to have low levels of total complement 1 year post reaction. Probable aggregates and fragments of human albumin (added stabilizer) were found in the streptokinase preparation and proved to be antigenic in some patients, but were not found to be related to the development of reactions. The findings suggest that patients who develop reactions to streptokinase cannot be predicted on the basis of antibody titres at presentation. Minor reactions to streptokinase would not appear to be antibody-mediated, although complement activation may be involved. PMID- 8222319 TI - Evaluation of in vivo immune complex formation and complement activation in patients receiving intravenous streptokinase. AB - The usefulness of several different methods for detecting immune complex formation and complement activation in the circulation were applied to samples from patients receiving intravenous Streptokinase therapy for myocardial infarction. Streptokinase is a foreign antigen and can cause immune reactions. We collected samples from 13 patients, before Streptokinase administration (baseline), at the end of infusion (1 h), 12 h later and on day 7. We measured IgG containing immune complexes (IgG-IC), free C3d and antibodies to Streptokinase by ELISA, and CR1, C3d and C4d on erythrocytes by flow cytometric assay. Antibodies to Streptokinase are common, as all but two of the patients had measurable antibody levels. During Streptokinase treatment there was a drop in antibody levels, most prominent in those patients who had high baseline levels. At the same time increased levels of free C3d and erythrocyte-bound C3d were observed. After 12 h free C3d was usually back to baseline level, but C3d on erythrocytes was still raised. These data indicate the formation of Streptokinase immune complexes in patients with high Streptokinase antibody levels, and show that these complexes are cleared rapidly from the circulation, leaving more persistent signs of complement activation. We conclude that free C3d is a good indicator of ongoing complement activation, whereas C3d on erythrocytes indicates that complement activation has recently taken place. PMID- 8222320 TI - Activation of human complement by mouse and mouse/human chimeric monoclonal antibodies. AB - The complement (C)-activating capabilities in human serum of 32 mouse and 10 mouse/human chimeric MoAbs of different isotypes, and their fragments, were tested in vitro. Activation of C via the classical pathway (CP) was performed in 1% factor D-deficient serum in gelatin containing Veronal buffer in the presence of calcium and magnesium (GVB++), while activation of the alternative pathway of C (AP) was assessed in 10% C1q-depleted serum in the presence of 5 mM MgCl2 in GVB++. The C-activating ability of MoAbs was expressed relative to the degree of activation of complement by aggregated IgG for the CP and relative to mouse IgG1 for the AP. All of seven mouse IgG2a MoAbs were potent activators of the CP. The results of CP activation by IgG1, IgG2b and IgG3 isotypes were different for individual MoAbs. Only three (two IgG1 and one IgG3) of 32 mouse MoAbs were potent activators of the AP. IgG2a and IgG2b were relatively poor AP activators. There were a few MoAbs which activated both the AP and CP. Of 10 chimeric MoAbs, two IgG1, one IgG2 and one IgG4 were poor or non-activators of the CP. On the other hand, IgG2 and IgG4 were good AP activators. IgG3 was the most potent AP activator. Most of the F(ab')2 fragments were activators of the AP and displayed no activation of the CP. Fc fragments only activated the CP, whereas Fab' did not activate the CP or the AP. These studies suggest that the route of complement activation by class and subclass MoAbs can not always be predicted in advance and based only on their subclass identity. PMID- 8222321 TI - Factors influencing the occurrence of active cytomegalovirus (CMV) infections after organ transplantation. AB - In this study, several factors influencing the occurrence of active CMV infection after organ transplantation (Tx) are analysed. For this purpose, 105 heart, kidney and lung transplant recipients who were CMV-positive or had a CMV-positive donor, were closely monitored for active CMV infection by antigenaemia, cultures, CMV serology and lymphocyte proliferation (LP) to CMV. Univariate and multivariate regression analysis were performed. As pretransplant risk factors the HLA-type and numbers of HLA mismatches between recipients and their donors, and the CMV serology of the recipient and donor were analysed. A new finding was that recipients of donors positive for HLA-B7 were especially at risk for developing active CMV infection (P = 0.03) and CMV disease (P = 0.03). This was not due to increased rejection treatment in these patients. Post-transplant risk factors for development of active CMV infection were absence of detectable cellular immunity to CMV (lymphocyte proliferation) after Tx (P < 0.01) and rejection treatment with OKT3 or ATG (P = 0.05). High levels of IgG anti-CMV did not prevent occurrence of active CMV infection or CMV disease in the CMV+ recipients. PMID- 8222322 TI - Diagnosis of typhoid fever: detection of Salmonella typhi porins-specific antibodies by inhibition ELISA. AB - Porins are highly immunogenic outer membrane proteins of Salmonella. Sera from typhoid patients contained a high level of IgG antibodies directed to porins of Salm. typhi. Since porins are highly conserved proteins, anti-porins antibodies both from typhoid patients and healthy normals reacted with porins from several Gram-negative bacteria. Therefore, in order to improve the specificity of detecting Salm. typhi porins-specific antibodies, an inhibition ELISA was developed using enzyme-conjugated MoAbs (MP1 and MPN4) specific to Salm. typhi porins. Sera from typhoid patients with positive haemoculture (16 out of 17) inhibited the binding of MP1 to porins, thus showing a positive test for typhoid, whereas sera from patients with other Gram-negative bacterial infections (n = 7) and from healthy volunteers (66 out of 67) were found to be negative. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of this assay were 94.1, 98.7, 97.8, 94.1 and 98.7% respectively. The validity of our inhibition ELISA for typhoid was higher than that of the Widal test. The diagnosis of typhoid fever as early as 3 days after the onset of fever, using a single specimen is possible. PMID- 8222323 TI - Distinct H-2 complex control of mortality, and immune responses to tuberculosis infection in virgin and BCG-vaccinated mice. AB - We have studied the impact of distinct haplotypes and of different alleles at specific H-2 loci on: (i) the susceptibility to lethal form of experimental tuberculosis; (ii) the level of DTH to mycobacterial antigens; (iii) the efficacy of vaccination with bacille Calmette-Guerin (BCG); and (iv) the IgG production and T cell proliferative response to H37Rv antigens. On the basis of median survival time (MST) following primary inoculation with lethal dose of Mycobacterium tuberculosis, susceptibility to infection associated with I-Ab and Db alleles, host resistance associated with I-Ak and Dd alleles. Mice bearing a disease-resistant phenotype also developed a vigorous DTH response. Vaccination with BCG before H37Rv infection significantly prolonged the survival time of both resistant and susceptible animals, except in B10.M (H-2f) mice. The latter exhibited intermediate resistance to infection before but slight decrease in the MST following a high-dose BCG vaccination. Distinct H-2 regulation of susceptibility to lethal infection and of BCG vaccination efficacy was confirmed in another relatively resistant H-2f-bearing strain A.CA, in which mortality occurred more rapidly in vaccinated compared with primarily infected animals. The expression of the H-2f haplotype was associated with a low DTH response to tuberculin following vaccination and subsequent lethal infection. The lack of BCG protection against Myco, tuberculosis challenge in B10.M mice associated with the high titre of specific IgG. In addition, these mice exhibited a unique ability to respond to 65-kD antigen by both IgG synthesis and T cell proliferation. PMID- 8222324 TI - Neutrophil activation in ivermectin-treated onchocerciasis patients. AB - Ivermectin is a safe and effective drug for onchocerciasis treatment. In certain individuals, however, therapy is accompanied by adverse reactions. The mechanisms underlying these reactions are not yet known. The aim of the present study was to investigate whether neutrophils are involved in the development of these adverse reactions. Elastase and lactoferrin, two markers for the release of neutrophil azurophilic and specific granule contents respectively, were measured by radioimmunoassays in plasma of onchocerciasis patients with varying degrees of side effects, as well as in control subjects before and 1 and 2 days after ivermectin treatment. A considerable increase of elastase levels after treatment was observed, whereas lactoferrin levels did not change. The percentage of patients with elevated elastase levels was significantly correlated with the degree of side effects. These findings suggest that neutrophil activation may be involved in the development of adverse reactions in these patients. PMID- 8222325 TI - Systemic immunization against IgA in immunoglobulin deficiency. AB - The presence of serum IgM and IgG antibodies against IgA is common among individuals with IgA deficiency. The route of immunization is still unknown, but it is possible that immunization occurs through the gut. We analysed anti-IgA antibody production in gastrointestinal lavage, saliva and breast milk from patients with IgA deficiency. In no case was there any evidence of local production of anti-IgA antibodies. Immunization may thus be due to exposure to endogenous IgA and therefore represent a 'true' autoimmune phenomenon which may possibly be involved in the pathogenesis of the disease. PMID- 8222326 TI - CD21-CD23 ligand pair expression in children with allergic asthma. AB - The CD23 antigen, a low affinity receptor for IgE, was recently shown to interact with another ligand, CD21, and the pairing of these molecules is important in T cell-B cell interaction and control of IgE production. Here, we analysed the expression of CD21 and CD23 on CD4+ and CD20+ lymphocytes in 25 allergic children and 12 age-matched non-allergic controls. Both the percentage (P < 0.01) and the absolute number (P < 0.001) of CD23+ cells were increased in allergic children. There was no difference of CD21+ cells. Double positive CD4+ CD23+ cells (2.5%) were only detected in one patient, in others all CD23 being expressed on B cells. The CD21 antigen was expressed only on B cells. Furthermore, allergic children had an increased mean fluorescence intensity of both the CD21 (P < 0.001) and the CD23 (P < 0.001) receptor. To analyse the possible difference in B cell subsets expressing CD21 and CD23 antigens, three-colour fluorescence analysis was performed. In allergic children the subset of CD20+ CD21- cells expressed more CD23 than in controls (P < 0.001). These results may mean an impaired expression and possibly regulation of CD21-CD23 interaction in allergic conditions. PMID- 8222327 TI - Multifunctional effects of anticomplementary agent K-76 on carrageenan-induced colitis in the rabbit. AB - In this study the effect of K-76, a sesquiterpene compound with anticomplementary activity isolated from a fungus culture, on carrageenan-induced colitis was studied from biochemical, histological and immunohistopathological aspects. K-76 suppressed epithelial cell loss, crypt abscess formation, inflammatory cell infiltration, mucosal atrophy, and ulceration. Immunohistochemical examination of the colonic mucosa showed that the number of IgG- and IgM-positive plasma cells and the staining intensity for IgG and C3 were increased in carrageenan-induced colitis, but these changes were inhibited by K-76. Besides, serum mucoprotein concentrations and CH50 levels were lower in the animals treated with carrageenan alone. K-76 exerted multifunctional activity, although its mechanisms of action remain obscure. PMID- 8222328 TI - Localization of tumour necrosis factor-alpha (TNF-alpha) and its receptors in normal and psoriatic skin: epidermal cells express the 55-kD but not the 75-kD TNF receptor. AB - The distribution of TNF-alpha, p55 TNF receptor (TNF-R) and p75 TNF-R in normal skin and uninvolved and lesional skin from psoriasis patients has been investigated, using specific mono- and polyclonal antibodies. In normal skin, and uninvolved and lesional skin from psoriasis patients, p55 TNF-R is associated with epidermal keratinocytes and a network of upper dermal dendritic cells. This suggests that the actions of TNF-alpha on epidermal cells in vivo are mediated by binding to the p55 TNF-R. In lesional psoriasis skin, there was staining of the parakeratotic stratum corneum and increased expression of p55 TNF-R in association with upper dermal blood vessels. Staining for p75 TNF-R in normal skin was restricted to eccrine sweat ducts and dermal dendritic cells, and was absent from the epidermis. In lesional psoriasis skin, there was staining for p75 TNF-R in association with upper dermal blood vessels and perivascular infiltrating cells. TNF-alpha in normal skin was predominantly localized to the basal cell layers of the epidermis, and was seen in association with eccrine ducts and sebaceous glands. In lesional psoriasis skin, and to a lesser extent in uninvolved psoriasis skin, TNF-alpha was distributed throughout the epidermis, and was also specifically localized to upper dermal blood vessels. Up-regulation of TNF-alpha, p55 TNF-R and p75 TNF-R on dermal blood vessels in psoriasis may play an important role in the pathogenesis of this condition by promoting cutaneous recruitment of inflammatory cells. PMID- 8222329 TI - Anti-CD28 antibody- and IL-4-induced human T cell proliferation is sensitive to rapamycin. AB - Rapamycin (RAPA) is a potent immunosuppressant. In this study we investigated the effect of RAPA on T cell proliferation triggered by various stimuli in an in vitro human model. The proliferation of T cells stimulated via an alternative pathway using phorbol myristate acetate (PMA) and anti-CD28 antibody (alpha CD28) in the absence of antigen-presenting cells (APC) was strongly inhibited by RAPA. T cell proliferation provoked via a combination of CD3/TCR and CD28 pathways using anti-CD3 antibody (alpha CD3) plus alpha CD28 was also inhibited by RAPA in the presence of APC. The mitogen (phytohaemagglutinin (PHA) or alpha CD3)-induced up-regulation of expression of the IL-2 receptor alpha chain (IL-2R alpha) and the IL-4 receptor (IL-4R) was sensitive to RAPA. This suggests that RAPA's interference with the IL-2 and IL-4 autocrine loops during T cell activation might contribute to RAPA's overall immunosuppressive effect. We have further demonstrated in a two-stage culture system that RAPA strongly inhibited IL-4 stimulated proliferation of T cells, the latter being either pretreated with alpha CD3 in the presence of APC, or with PMA plus alpha CD28 in the absence of APC. The result suggests that the Ca++ influx during the pretreatment is not obligatory for T cells to achieve IL-4 responsiveness. The results also indicate that RAPA's antiproliferative effect on IL-4-stimulated T cells is not contingent on the various mechanisms of cell priming. Therefore, RAPA's major target is probably at the second stage after the priming. Our study has extended current knowledge about the effect of RAPA on human T cells. PMID- 8222330 TI - Antigen-presenting capacity of macrophages and dendritic cells in the peritoneal cavity of patients treated with peritoneal dialysis. AB - In this study the antigen-presenting capacity of human peritoneal cells and the influence of continuous ambulant peritoneal dialysis (CAPD) were studied. On average 6% of the peritoneal cells were dendritic cells (DC), with no difference between CAPD and control peritoneal cells. DC were enriched by selecting for non adherent, Fc receptor-negative, low density cells. A typical spot-like CD68 positivity was seen in DC, in contrast to the pancytoplasmic staining pattern in macrophages. Peritoneal DC morphologically and functionally showed features of cells belonging to the DC lineage. Peritoneal DC were superior antigen-presenting cells for both allo-antigen, and Candida albicans antigen or purified protein derivative. CAPD peritoneal macrophages were two- to three-fold better stimulator cells for allogeneic T cells compared with control macrophages. The level of integrins/adhesins or MHC class I or II, as measured semi-quantitatively on the FACS, could not account for this phenomenon. In addition, a double chamber system showed that dialysate-activated macrophages produced soluble factors that could enhance DC-induced allogeneic T cell proliferation. In conclusion, human peritoneal cells contain a relatively high percentage of classical DC. CAPD treatment does not impair the antigen-presenting capacity of peritoneal cells, but instead upregulates the allo-antigen-presenting capacity of peritoneal macrophages. PMID- 8222332 TI - Nomenclature for T cell receptor (TCR) gene segments of the immune system. WHO IUIS Nomenclature Sub-Committee on TCR Designation. PMID- 8222331 TI - Characterization of a human monoclonal autoantibody directed to cardiolipin/beta 2 glycoprotein I produced by chronic lymphocytic leukaemia B cells. AB - We determined the specificity and sequence of immunoglobulin molecules synthesized by monoclonal B cells from a patient with chronic lymphocytic leukaemia (CLL) who presented with a number of clinical and biological autoimmune symptoms. Heterohybrids obtained by fusion of CLL cells with the mouse X63-Ag 8.653 myeloma produced IgM lambda MoAbs directed to the cardiolipin/beta 2 glycoprotein I (beta 2GPI) complex and ssDNA. They were devoid of polyreactivity. Nucleotide sequence analysis of the variable domain of the mu chain indicated the utilization of the VH4 71.2 gene or one allotypic variant, DXP4 and JH3 segments. The lambda light chain used the single gene from the V lambda 8 subfamily, J lambda 3 and C lambda 3 genes. The VH gene displayed 11 nucleotide changes in comparison with its putative germline counterpart. However, these nucleotide changes correspond to variations observed in other published VH4 sequences, suggesting gene polymorphism rather than somatic mutation. DXP4 and JH3 were also in germline configuration. The VL gene exhibited a single replacement mutation in CDR1. These data suggest that the monoclonal CLL B cells in this patient retained VH and VL genes in germline configuration although they secreted a pathogenic anti-cardiolipin antibody associated with clinical symptoms, vasculitis and thrombosis, which may be provoked by antibodies to the phospholipid/beta 2GPI complex. PMID- 8222333 TI - The inhibitory effect of SA3443, a novel cyclic disulfide compound, on alpha naphthyl isothiocyanate-induced intrahepatic cholestasis in rats. AB - 1. We investigated the effect of SA3443, a novel cyclic disulfide compound, on alpha-naphthyl isothiocyanate (ANIT)-induced intrahepatic cholestasis in rats. 2. SA3443 given orally at doses of 100 or 300 mg/kg, three times at 4h intervals, dose-dependently suppressed the elevation of serum total bilirubin, alkaline phosphatase activity and transaminase activities induced by administration of ANIT (80 mg/kg) 3h before the second dosages of SA3443. 3. SA3443 also significantly inhibited the reduction of bile flow in the ANIT-treated rats. 4. These findings indicated that SA3443 had a markedly inhibitory effect on ANIT induced intrahepatic cholestasis in rats. PMID- 8222334 TI - Use of a computerized postural sway measurement system for assessing workers exposed to manganese. AB - 1. Computerized postural sway measurement systems have been used recently in human physiology and pharmacology for determining postural stability, but their applicability for the assessment of the effects of exposure to neurotoxic agents is lacking. 2. We have examined the feasibility of using a computerized postural sway measurement system to assess the postural sway parameters of manganese exposed workers compared with a control group. 3. Sway parameter data were collected using a KISLTER multicomponent measuring platform (Type 9281B) connected to VICON motion analysis system for 13 exposed and 16 control subjects. 4. Significant differences in several of the sway parameters (Px, Py--mean distance (mm), from the centre of the platform along the X-axis [anterior posterior movement] and Y-axis [lateral movement], respectively) between the exposed and control groups were observed even after adjustment for possible confounders. Computerized postural sway measurement system may be a useful method of assessing workers exposed to neurotoxic agents affecting posture. PMID- 8222335 TI - Sheep hypothalamus contains a non-angiotensin ligand for type 1 and type 2 angiotensin II receptors. AB - 1. The aim of this study was to determine whether the brain contains an alternative ligand for angiotensin II (AII) receptors. 2. A radioreceptor assay based upon bovine cerebellar membranes (Type 2 AII receptors) was used to monitor the partial purification of an AII-like material from sheep hypothalami. 3. This material displaces 125I-[Sar1, Ala8]-AII from both type 1 (rat adrenal capsular membranes) and Type 2 AII receptors in a manner parallel to that of AII. It has a size of approximately 30,000 Da, is strongly cationic, is stable to boiling but is destroyed by trypsin. It is not recognized by AII antisera. 4. These data provide direct evidence for a non-angiotensin endogenous ligand for brain AII receptors. This novel ligand may play a role in the regulation of blood pressure and other actions mediated by brain AII receptors. PMID- 8222336 TI - Demonstration of defective glucose uptake and storage in erythrocytes from non insulin dependent diabetic patients and effects of metformin. AB - 1. Red blood cells can store glucose and may thus participate in blood glucose homeostasis. We investigated if a defect in this process exists in non-insulin dependent diabetes (NIDD). 2. Blood was obtained in fasting conditions from 10 normal and 10 newly diagnosed NIDD patients (before and after 4 weeks Metformin therapy). Washed erythrocytes were resuspended in media containing various glucose concentrations (4.4, 6.6, 8.8 and 13.2 mmol/L). Total glucose uptake was calculated as the sum of the measurements of lactate as well as free glucose, the latter being determined before and after addition of amyloglucosidase to the pellet. 3. Cells from diabetics showed a pronounced reduction in glucose uptake, particularly in their capacity to store glucose as glycogen (reactive to amyloglucosidase). Metformin treatment almost normalized glycogen levels, whereas lactate declined concomitantly in the pellet. 4. Our data demonstrate that a defect in glucose uptake exists in erythrocytes from NIDD patients, affecting both free and stored glucose, and that this defect is reversed by Metformin treatment, indicating that this drug can increase glycogen levels even in insulin insensitive cells. 5. Thus, in view of their total mass, erythrocytes may be important in the impaired glucose homeostasis in NIDD, in particular in marked hyperglycaemia such as after a meal. PMID- 8222337 TI - Effect of arginine vasopressin and parathyroid hormone-related protein on renal function in the ovine foetus. AB - 1. The effects of intravenous infusions of arginine vasopressin (AVP), parathyroid hormone-related protein (PTHrP) and AVP+PTHrP on renal function in intact ovine foetuses at 100-125 days of gestation were examined. 2. A low dose of AVP (5.5 +/- 0.9 pmol/h) increased plasma AVP concentrations from 0.6 pmol/L to 2.1 +/- 0.4 pmol/L (mean +/- s.e.m; n = 8). This dose caused a significant reduction in free water clearance (CH2O; P < 0.001), without any significant change in fetal arterial blood pressure, glomerular filtration rate (GFR), or the urinary excretion rates of sodium, calcium or 3',5'-cyclic adenosine monophosphate (cAMP). 3. Infusions of PTHrP (1 nmol/h), with or without 1 nmol bolus dose, significantly increased (P < 0.05) urine osmolality (UOSM), but did not synergize with AVP in reducing CH2O. 4. It is concluded that AVP and PTHrP do not act synergistically on the kidney of the intact ovine foetus. PMID- 8222338 TI - Effect of xanthine derivatives on chemotactic polypeptide-induced superoxide and enzyme release from human polymorphonuclear leucocytes. AB - 1. We investigated the effects of new xanthine derivatives, 1-methyl-3-propyl xanthine (MPX) and 1,3-dipropyl xanthine (DPX), and several other xanthine derivatives on N-formyl-methionyl-leucyl-phenylalanine-induced superoxide and lysozyme release from human polymorphonuclear leucocytes (PMN). 2. MPX and DPX at low concentrations (10(-8) - 10(-9) mol/L) inhibited superoxide release from PMN by a maximum of 31.2 +/- 10.6% and 49.8 +/- 10.4% (mean +/- s.d.), respectively, and 10(-3) mol/L concentrations completely inhibited the release reactions (4.8 +/- 1.2 and 7.6 +/- 2.5% of control level). At 10(-5) mol/L, however, the inhibition did not occur (99.9 +/- 7.3 and 110.2 +/- 15.8% of control level). When PMN was pre-incubated with adenosine deaminase (ADA, 0.1 U/mL), superoxide release from PMN was inhibited in a dose-dependent manner by MPX and DPX and the interruption of the inhibition at 10(-5) mol/L was not observed. 3. Lysozyme release from PMN was inhibited by MPX at low concentrations (10(-7) - 10(-6) mol/L) and high concentrations (10(-3) mol/L). However 10(-4) mol/L of MPX facilitated the release (23.7 +/- 27.0%). When pretreated with ADA (0.1 U/mL), MPX suppressed lysozyme release in a dose-dependent manner and the facilitation of the release at 10(-4) mol/L was not observed. 4. When comparing effects of some other xanthine derivatives on superoxide release, the interruption of the inhibition of superoxide release at 10(-5) mol/L was commonly observed among xanthine derivatives with adenosine A2 antagonism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222339 TI - Intralymphocytic free calcium and magnesium in stroke-prone spontaneously hypertensive rats and effects of blood pressure and various antihypertensive agents. AB - 1. Free Ca2+ ([Ca2+]i) and Mg2+ ([Mg2+]i) were measured in peripheral lymphocytes from stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar Kyoto rats (WKY) at the age of 5, 7 and 17 weeks, from various antihypertensive agents-treated SHRSP, and from secondary hypertensive WKY. 2. At the age of 5 weeks, no difference was observed in systolic blood pressure (SBP), or lymphocyte [Ca2+]i and [Mg2+]i between SHRSP and WKY. At the age of 7 or 17 weeks, SBP and [Ca2+]i of SHRSP were significantly higher than in WKY, and at the age of 17 weeks, [Mg2+]i of SHRSP was significantly lower than in WKY. Further, [Ca2+]i or [Mg2+]i was positively or negatively correlated to SBP, and [Mg2+]i was negatively correlated to [Ca2+]i. 3. SBP of SHRSP fell significantly after antihypertensive treatment with calcium antagonist, angiotensin-converting enzyme (ACE) inhibitor or hydralazine for 40 days. [Ca2+]i was significantly lower in calcium antagonist and hydralazine groups, and tended to be low in ACE inhibitor group. These four groups showed no difference in [Mg2+]i. 4. After 40-day administration of NG-nitro-L-arginine (L-NNA), WKY developed severe hypertension, but there were no significant differences in lymphocyte [Ca2+]i and [Mg2+]i between the L-NNA treated and non-treated groups. 5. These results suggested that increased lymphocyte [Ca2+]i and decreased [Mg2+]i observed in SHRSP are not only secondary to hypertension but possibly related to a basic genetic abnormality of divalent cation handling. PMID- 8222340 TI - Effect of nicorandil on post-ischaemic contractile dysfunction in the heart: roles of its ATP-sensitive K+ channel opening property and nitrate property. AB - 1. This study aimed to characterize the effect of nicorandil (NC) on myocardial stunning and the role of ATP-sensitive K+ (KATP) channel opening property in its cardioprotective action. 2. In open-chest anaesthetized rabbits, myocardial stunning was induced by 10 min of coronary occlusion followed by 30 min of reperfusion. As an index of regional contractile function, systolic thickening fraction (TF) was measured by an epicardial Doppler sensor. The doses of NC (10 micrograms/kg per min) and nitroglycerin (TNG) (1 micrograms/kg per min) were selected not to lower the systemic blood pressure significantly. 3. In the untreated controls, TF at 30 min after reperfusion was 46.4 +/- 2.9% of the baseline value, indicating myocardial stunning. Both NC and TNG significantly improved post-ischaemic recovery of TF when administered during the pre-ischaemic and post-ischaemic periods (TF = 68.2 +/- 6.4%, 64.7 +/- 2.3%, respectively). However, when their infusion was restricted to a pre-ischaemic 10 min period, TF recovery was improved by NC, but not by TNG (63.4 +/- 7.9%, 40.9 +/- 6.2%, respectively). 4. Pretreatment with glibenclamide (GL; 0.3 mg/kg) did not influence the recovery of TF after the 10 min ischaemia (TF = 52.4 +/- 3.9% at 30 min after reperfusion). However, after the GL injection, a cardioprotective effect from nicorandil pretreatment was not detected (TF = 51.3 +/- 1.7%). 5. These results suggest that nicorandil protects the myocardium against stunning by opening the KATP channel when it is given before ischaemia, and that the nitrate property of nicorandil may also play a role during the reperfusion period in attenuation of post-ischaemic contractile dysfunction. PMID- 8222341 TI - Release of a substance from the human placenta having digoxin-like immunoreactivity. AB - 1. The human placental lobule, perfused with a constant flow (5 mL/min) of Krebs' solution after delivery at term, released into the fetal perfusate a digoxin-like substance, as measured by a fluorescence polarization immunoassay. 2. Initially the venous concentration was 360 +/- 66.7 pmol/L digoxin equivalents. This level did not change significantly during fetal vasoconstriction induced by prostaglandin F2 alpha infusion and persisted for the duration of the experiment (1.5-2 h). 3. Infusion into the fetal circulation of Fab fragments of sheep antibodies to digoxin caused vasodilatation, indicated by a fall in perfusion pressure. 4. Thus a digoxin-like immunoreactive substance, previously reported to be present in the placenta, is released into the fetal circulation and may play a role in placental control of fetal vascular tone. PMID- 8222342 TI - The use of antisense approaches to study development. PMID- 8222343 TI - Creating a conditional mutation of Wnt-1 by antisense transgenesis provides evidence that Wnt-1 is not essential for spermatogenesis. AB - We have used mice transgenic for an antisense construct for Wnt-1 to study the role of this gene in post-meiotic sperm development. The human PGK-2 promoter provided levels of Wnt-1 antisense mRNA in testes in 5 transgenic lines greatly in excess of Wnt-1 mRNA concentrations, and Wnt-1 mRNA levels were greatly decreased in the lines, by 98% in three of them. There was a general correlation between copy number of the insert, levels of antisense RNA, and decreases in mRNA. There was little effect of the antisense transgene on fertility or testicular histology suggesting that normal levels of Wnt-1 transcript are not essential for spermatogenesis. PMID- 8222344 TI - Use of a tomato mutant constructed with reverse genetics to study fruit ripening, a complex developmental process. AB - Fruit ripening is one of the most dramatic developmental transitions associated with extensive alteration in gene expression. The plant hormone ethylene is considered to be the causative ripening agent. Transgenic tomato plants were constructed expressing antisense or sense RNA to the key enzyme in the ethylene (C2H4) biosynthetic pathway, 1-aminocyclopropane-1-carboxylate (ACC) synthase using the constitutive CaMV 35S and fruit specific E8 promoters. Fruits expressing antisense LE-ACS2 RNA produce less ethylene and fail to ripen only when ethylene production is suppressed by more than 99% (> 0.1 nl/g fresh weight). Ethylene production is considerably inhibited (50%) in fruits expressing sense LE-ACS2 RNA. Antisense fruits accumulate normal levels of polygalacturonase (PG), ACC oxidase (pTOM13), E8, E17, J49, and phytoene desaturase (D2) mRNAs which were previously thought to be ethylene-inducible. E4 gene expression is inhibited in antisense fruits and its expression is not restored by treatment with exogenous propylene (C3H6). Antisense fruits accumulate PG mRNA, but it is not translated. Immunoblotting experiments indicate that the PG protein is not expressed in antisense fruits but its accumulation is restored by propylene (C3H6) treatment. The results suggest that at least two signal-transduction pathways are operating during tomato fruit ripening. The independent (developmental) pathway is responsible for the transcriptional activation of genes such as PG, ACC oxidase, E8, E17, D2, and J49. The ethylene-dependent pathway is responsible for the transcriptional and posttranscriptional regulation of genes involved in lycopene, aroma biosynthesis, and the translatability of developmentally regulated genes such as PG. PMID- 8222345 TI - Opposite functions of jun-B and c-jun in growth regulation and neuronal differentiation. AB - Induction of the jun-B and/or c-jun transcription factors is part of the immediate early response to diverse stimuli that induce alterations in cellular programs. While c-jun is a protooncogene whose expression is required for induction of cell proliferation, jun-B has recently been found to be induced by stimuli inducing differentiation in various cell lines. Furthermore, its expression is largely restricted to differentiating cells during embryogenesis. To determine the functional significance of these findings, we used antisense phosphorothioate oligodeoxynucleotides to inhibit expression of the two genes in proliferating and neuronally differentiating cells. While selective inhibition of c-jun expression reduced proliferation rates, inhibition of jun-B protein synthesis markedly increased proliferation in 3T3 fibroblasts, human mammary carcinoma cells and PC-12 pheochromocytoma cells, suggesting jun-B involvement in negative growth control. Neuronal differentiation of PC-12 cells induced by nerve growth factor (NGF) was prevented by inhibition of jun-B protein synthesis. PC-12 cells not only failed to grow neurites but also remained in the proliferative state. Furthermore, in cultured primary neurons from rat hippocampus, inhibition of jun-B expression, again, markedly reduced morphological differentiation. Conversely, inhibition of c-jun protein synthesis enhanced morphological differentiation of both primary neurons and PC-12 tumor cells. Thus, jun-B expression is required for neuronal differentiation and its balance with c-jun activity is involved in regulating key steps in proliferation and differentiation processes. PMID- 8222346 TI - Regulation of the Dictyostelium glycogen phosphorylase 2 gene by cyclic AMP. AB - A crucial developmental event in the cellular slime mold, Dictyostelium discoideum, is glycogen degradation. The enzyme that catalyzes this degradation, glycogen phosphorylase 2 (gp-2), is developmentally regulated and cAMP appears to be involved in this regulation. We have examined several aspects of the cAMP regulation of gp-2. We show that addition of exogenous cAMP to aggregation competent amoebae induced the appearance of gp-2 mRNA. The induction of gp-2 mRNA occurred within 1 and 1.5 h after the initial exposure to cAMP. Exposure to exogenous cAMP concentrations as low as 1.0 microM could induce gp-2 mRNA. We also examined the molecular mechanism through which cAMP induction of gp-2 occurs. Induction of gp-2 appears to result from a mechanism that does not require intracellular cAMP signaling, and may occur directly through a cAMP binding protein without the requirement of any intracellular signalling. We also examined the promoter region of the gp-2 gene for cis-acting elements that are involved in the cAMP regulation of gp-2. A series of deletions of the promoter were fused to a luciferase reporter gene and then analyzed for cAMP responsiveness. The results indicated that a region from -258 nucleotides to the transcriptional start site is sufficient for essentially full activity and appears to carry all necessary cis-acting sites for cAMP induction. Further deletion of 58 nucleotides from the 5' end, results in fivefold less activity in the presence of cAMP. Deletion of the next 104 nucleotides eliminates the cAMP response entirely. PMID- 8222347 TI - The wound healing process. AB - The field of wound repair continues to grow at an astounding pace. In this review we have discussed both the basic science and the clinical aspects of wound healing. The basic science section details the three phases of wound healing. The inflammatory phase is marked by platelet accumulation, coagulation, and leukocyte migration. The proliferative phase is characterized by re-epithelialization, angiogenesis, fibroplasia, and wound contraction. Finally, the remodeling phase takes place over a period of months, during which the dermis responds to injury with the production of collagen and matrix proteins and then returns to its preinjury phenotype. The second part of this review outlines selected practical clinical aspects of wound repair. That section is subdivided into acute and chronic wounds. We describe the difference between primary and secondary intention healing and discuss factors that affect healing in both. We also describe our systemic approach to dealing with patients with chronic wounds and present our problem-oriented treatment program based on the cause of the chronic wound. PMID- 8222348 TI - Re-epithelialization. Human keratinocyte locomotion. AB - Keratinocyte migration and proliferation both play a role in covering skin wounds by the process of re-epithelialization. Connective tissue components are a powerful influence on keratinocyte locomotion. The mechanisms of keratinocyte locomotion and cellular division are independent. Both connective tissue components and soluble factors may serve to enhance keratinocyte migration. In addition to the field of growth factors, we would like to suggest that there should be the recognition of an entire new class of agents called migration factors. PMID- 8222349 TI - Biology of dermal wound repair. AB - Dermal wound repair events can be temporally, albeit arbitrarily, grouped into inflammation, granulation tissue formation, and tissue remodeling. These phases are not mutually exclusive but rather overlap in time. This review follows the chronology of dermal wound repair, that is, inflammation, granulation tissue formation, and tissue remodeling. PMID- 8222350 TI - Growth factors and wound healing. AB - If one looks at the development of most drugs, it becomes evident that an extended period of time is necessary before the proper use and full range of activities of the drug in treating human disease become apparent. In many cases, initial treatment failures are due to faulty expectations as well as inadequate dosing and delivery systems. It is my opinion that growth factor therapy will follow a similar course but that it ultimately will have a role in the modulation of wound repair. PMID- 8222351 TI - Fetal tissue repair and wound healing. AB - The fibrosis and scar formation that characterize adult wound healing are also the cause of clinical problems; scar contracture, hypertrophic scar, and pulmonary and hepatic fibrosis are only a few examples. Studies of fetal wound healing can provide an insight into the initiation and regulation of a scarless repair process akin to regeneration. Studies of fetal repair have already suggested mechanisms that might favorably alter adult healing. Topical application of hyaluronic acid to wounds in adult diabetic rats leads to enhanced epithelial migration. It has been recognized that the addition of TGF-beta to fetal wounds causes an adultlike healing response with fibrosis and inflammation. A subsequent study using neutralizing antibody to TGF-beta in adult wounds showed enhanced healing with a more normal dermal architecture with fewer macrophages, fewer blood vessels, and less collagen. As our understanding of regenerative tissue repair increases, the opportunities to modulate adult fibrotic conditions should expand. PMID- 8222352 TI - Mast cell and myofibroblast in wound healing. AB - Growing evidence in the literature indicates that mast cells are integrally involved in the process of dermal wound repair. They are resident cells of the normal dermis and have several cytokines stored in their granules that are stimulatory to fibroblasts. They also contain serine proteases that may be involved in remodeling of the extracellular matrix during healing. Mast cells are found in increased numbers in acute wounds and in certain chronic fibrotic diseases. Their influence on fibroblast growth and collagen production may be an important element in fibrosis. The effects of mast cell mediators on dermal fibroblasts are currently being explored by our laboratory and others. Myofibroblasts are implicated in the phenomenon of wound contraction. These phenotypically altered fibroblasts express some features of smooth muscle cells, notably actin filaments, and are abundant in granulation tissue. It has been proposed that they are responsible for wound contraction and possibly certain types of contracture. However, this hypothesis has been challenged by studies demonstrating the presence of myofibroblasts in wounds that do not contract, or the process of contraction in vitro in the absence of myofibroblasts. At this time the issue remains open to debate and further research. PMID- 8222353 TI - Scars and keloids. AB - Despite the disfigurement and symptoms from these abnormal wound responses, the literature offers little consensus about appropriate therapy. Few prospective blinded studies have been performed to compare therapeutic options. Multiple therapies have been reported. Most physicians agree with Kelly's emphasis on the importance of prevention. All effort should be directed to avoid or to minimize any stimuli that could promote an abnormal wound response in predisposed individuals. Treatment must be carefully selected for appropriate lesions. Hypertrophic scars may be surgically corrected, but other measures such as intralesional corticosteroid injection, pressure therapy, cryotherapy, and other topical treatments may be useful. These scars have a different clinical course than do keloids. Published recurrence rates for keloid treatment are lowest for the combination of surgery and radiation. Although some keloids may respond to intralesional steroid injection, keloid tissue must be removed and the resulting wound must be treated to minimize the risk of recurrence. PMID- 8222354 TI - Management of burn wounds. AB - The pathophysiology of thermal injury is described and applied to the clinical diagnosis of the depth of the injury. The basis of the selection of the type of nonoperative therapy for moderate depth second-degree burns is outlined for optimizing healing time. The importance of early tangential (sequential) excision of deep second-degree burns and third-degree burns in reducing functional and cosmetic residual is emphasized. PMID- 8222355 TI - Unique wounds and wound emergencies. AB - Unique wounds and wound emergencies include a variety of environmental assaults including bites, stings, electrical and chemical injury, and severe reactions to medications. This article reviews the clinical manifestations and treatment options essential to the management of these environmental assaults. PMID- 8222356 TI - Drug and nutrient aspects of wound healing. AB - An understanding of the major factors that influence wound healing enables the clinician to better prepare patients for surgery and to identify and correct the effects of either drugs or nutritional imbalances in patients whose wounds fail to heal. Ideally, factors that enhance wound healing should be optimized, and detrimental influences minimized whenever possible. Among the major factors influencing wound healing are the state of general health, drugs, and nutritional status. Nutritional assessment prior to surgery, following injury, and during recovery is an important part of management of patients with extensive wounds. In dermatologic surgical practice, nutritional support is not indicated; however, subclinical nutritional deficiencies may not be uncommon and may become relevant if wound healing is significantly impaired. Dermatologic surgeons must be aware of the potential for many drugs to interfere with wound healing and interrupt therapy prior to surgery whenever possible. Some drugs cannot be stopped and surgery should be performed only when absolutely necessary when patients are receiving antineoplastic, anticoagulant, and high-dose steroid therapy. PMID- 8222357 TI - Wound healing microbiology. AB - The relationship between wound healing and cutaneous microbiology is a complex one mediated by the type of wound, its treatment, and the defense mounted by the injured individual. Acute wounds harbor microflora similar to that of noninjured skin. "Dirty" traumatic wounds or chronic wounds with devitalized tissue offer more opportunity for microflora colonization, with the potential for bacterial adherence to the host cells and ultimate invasion into viable tissue or infection. The alterations of these wound environments can be brought on by topical treatments. Occlusive dressings with various moisture vapor transmission rates can provide a wound environment suitable for microflora proliferation. In spite of this increase in numbers of organisms, most wounds do not become infected, as shown by a survey of published trials. Infection requires the proper pathogenic microorganism, its attachment, and subsequent multiplication. To cause an infection, an organism must invade viable tissue and evade the host defense response. The many occlusive dressing types available offer an opportunity to select a wound therapy tailored for the microenvironment most suitable for healing. Wounds can be protected from exogenous microfloral contamination with dressings. Some dressing types, such as hydrocolloid dressings with no moisture vapor transmission, should not be used on chronic wounds suspected of harboring anaerobic organisms. Other typical treatments, such as antiseptics, can injure tissue and make infection more probable, providing devitalized tissue for bacterial adhesion. Povidone-iodine solution has been shown not be efficacious, whereas povidone-iodine cream effectively limits bacteria and infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222358 TI - Wound healing and aging. AB - Age-related differences in wound healing have been clearly documented. Although the elderly can heal most wounds, they have a slower healing process, and all phases of wound healing are affected. The inflammatory response is decreased or delayed, as is the proliferative response. Remodeling occurs, but to a lesser degree, and the collagen formed is qualitatively different. Diseases that affect wound healing are more prevalent in the elderly and have a greater adverse effect on healing than in young adults. Thus, particularly in the elderly, concomitant medical problems should be treated vigorously to allow for maximum healing. Recent trials of novel therapies to enhance wound healing suggest, however, that much can be done to improve the prognosis of elderly patients with risk factors known to adversely affect wound healing. PMID- 8222359 TI - Management of acute wounds. AB - Acute wounds are defined as a disruption in the integrity of the skin, including the epidermis and dermis. Healing is the reparative process by the skin to protect itself and to generate new cutaneous structures. Medical and surgical intervention in wounds can greatly affect the resultant scar and the time of healing. PMID- 8222360 TI - Management of chronic and problem lower extremity wounds. AB - With simple, inexpensive therapy, virtually all venous ulcers heal without the need for hospitalization, skin grafting, growth factors, hyperbaric oxygen, or debilitating bed rest. Although the future promises to yield a greater understanding about the nature of these wounds and ways to treat them, one can benefit the vast majority of patients with available technology. Few therapeutic experiences are as gratifying as healing a patient's debilitating chronic wound. PMID- 8222361 TI - Electrical stimulation and ultrasound in wound healing. AB - The events that lead to tissue repair are very complex. Because our understanding of these processes is increasing in scope, the use of nontraditional treatment therapies should be considered. Evidence is reported in the literature that both electrical stimulation and ultrasound therapies may be beneficial in certain circumstances to heal various wound types. Owing to clinicians' unfamiliarity with the current research and general understanding of such therapies, many patients receive only traditional treatment and remain unexposed to the potential benefits of the nontraditional. With continued research to better define optimal treatment parameters, improved wound healing will result. PMID- 8222362 TI - Lasers and wound healing. AB - The advances in laser surgery over the past decade have been remarkable and have significantly altered the management of a host of dermatologic disorders. This article focuses on the CO2 laser as a cutting and vaporization tool and reviews the features that distinguish CO2 laser-induced wounds from those created by a scalpel. Tissue welding, wound management after skin treatment with the visible light lasers based on the principle of selective photothermolysis, as well as the controversial field of low-energy laser therapy (biostimulation) are also addressed. PMID- 8222363 TI - Wound healing in the developing world. AB - Most would recognize that nonhealing is due to malnutrition, infection, or failure of blood supply, but in practice in a world in which there is now much understanding of how to manage nonhealing, the real issue is how to direct this knowledge so that it becomes available to all. There is a failure of delivery of care. When considering quality of life issues, people's preferences, priorities, and motivation have to be taken into account, but this can be applied also to the deliveries of care. Caretakers must want to relieve odor and pain and help the mobility of their patients, encouraging them to be well groomed, allowing them to maintain adequate hygiene by bathing and, above all, making them welcome so that they can be embraced, can marry, and can be employed. To achieve these goals, complete healing is necessary with normal color, contour, and function. Management of wounds in the third world requires missionary zeal for some basic objectives. Nothing can be taken for granted. There must be food and drink for the patient, protection from flies, antisepsis, good surgical technique, a temperate environment, and a knowledge of the requirements for achieving one's potential in each and every community. PMID- 8222364 TI - Treatment of oral ulcers. AB - Oral ulcers may be the result of allergic, infectious, metabolic, nutritional, or malignant systemic disorders or a variety of other inflammatory diseases. These range from minor irritations to signs of life-threatening illnesses. Patients with oral ulcers should adhere to a variety of general measures that are recommended plus specific treatments for specific diagnoses. PMID- 8222365 TI - The requirement of low calcium dialysate in patients on continuous ambulatory peritoneal dialysis receiving calcium carbonate as a phosphate binder. AB - In the present study we investigated the requirement of low calcium dialysate in 35 patients on continuous ambulatory peritoneal dialysis (CAPD) receiving calcium carbonate as the sole phosphate binder over a 12-month period. Patients with corrected serum calcium > or = 2.85 mmol/L after switching to oral calcium carbonate were given 1 to 3 2-litre exchanges of 2.5 mEq/L calcium dialysate. Serum phosphate level dropped from the pretreatment value of 2.95 +/- 0.62 to a level of between 1.70 +/- 0.41 to 2.03 +/- 0.44 mmol/L 2 weeks after therapy. Corrected serum calcium level increased significantly from 2 weeks onwards. Serum alkaline phosphatase rose initially at 2 and 6 weeks and decreased from 3 months onwards. Serum parathyroid hormone level dropped significantly from a mean pretreatment level of 569 to 320 pg/ml after 12 months (p < 0.001). Serum aluminum decreased significantly from a mean of 1.04 to 0.65 umol/L (p < 0.01). Daily calcium carbonate requirement fluctuated but tended to increase till 8 months and plateaued and ranged from 2.61 +/- 0.57 to 3.98 +/- 2.11 gm. The daily requirement of low calcium dialysate followed a similar trend with approximately three-quarters of patients ultimately requiring at least 1 bag of low calcium dialysate. Eight patients did not require low calcium dialysate. Patients who required low calcium dialysate were significantly older, had a significantly lower pretreatment serum parathyroid hormone and higher serum aluminum levels than those who did not.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222366 TI - Effect of dialysis on the clearance of Staphylococcus epidermidis from the peritoneal cavity: an experimental evaluation. AB - Current information on the effect of peritoneal dialysis on host defenses in the peritoneal cavity is limited to in vitro studies. In the present study, clearance of Staphylococcus epidermidis from the abdominal cavity was assessed in an animal model undergoing peritoneal dialysis. In non-manipulated subjects, challenges of up to 10(8) viable microorganisms were cleared from the peritoneal cavity over a period of 24-48 hours. Cannula implantation, short- and long-term dialysis and the presence of excessive amounts of fluid in the peritoneal cavity did not affect the clearance rate. Presentation of the bacterial challenge, either as a bolus or via a slow delivery system, likewise had no effect on clearance rates. The results suggest that microbial trophism and the characteristics of the invading pathogen may be more critical determinants of the outcome of an infectious challenge to the peritoneal cavity than the status of the cellular and humoral defense systems. PMID- 8222367 TI - A drug-induced turbid peritoneal dialysate in five patients treated with continuous ambulatory peritoneal dialysis. AB - Turbid peritoneal dialysate is one of the heralding signs for infective peritonitis in patients treated with continuous ambulatory peritoneal dialysis (CAPD). Peritoneal dialysate drained from 5 out of 8 patients undergoing CAPD became turbid within 24 h after the administration of a new dihydropyridine type of calcium channel blocker, manidipine hydrochloride (10 or 20 mg/day). Although the dialysate was visually indistinguishable from that observed during infective peritonitis, no clinical manifestations being suggestive of infective peritonitis were observed. The turbid dialysate contained normal leukocyte counts (< 10 mm 3), and the bacterial (aerobic and anaerobic) and fungal cultures of the dialysate failed to produce organisms. Cytology of the dialysate showed no malignant cells. Biochemical analysis of the dialysate revealed that the fluid contained an elevated triglyceride concentration [range: 12 to 32 mg/100 ml (0.14 to 0.37 mmol/l], while the clear dialysate obtained from 9 uncomplicated CAPD patients contained less than the detection limit of the assay for triglyceride [< 5 mg/100 ml (0.06 mmol/l)]. All patients were found to consume a nutritionally balanced diet consisting of 1800 to 2400 Kcal/day during the manidipine therapy. No appreciable change was observed in the mean (+/- SD) fasting serum triglyceride concentrations determined before and 24 h after the withdrawal of the manidipine therapy (194 +/- 84 and 186 +/- 106 mg/100 ml, respectively) in the 5 CAPD-patients with turbid peritoneal dialysate. Within 24 h after the withdrawal of the manidipine therapy the peritoneal dialysate became clear spontaneously and the triglyceride concentration in the dialysate was normalized in all of them.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222368 TI - Serum creatinine concentration in chronic hemodialysis patients: higher predialytic levels in black males. PMID- 8222369 TI - Successful anticoagulation treatment in an anuric patient with thrombosis of the suprarenal segment of the inferior vena cava after right nephrectomy. PMID- 8222370 TI - Vasculitis involving lung, toes and kidneys in a patient with angioedema: a common pathogenesis? PMID- 8222371 TI - PCR and hemodialysis patients. PMID- 8222372 TI - Renal function and kidney morphology in biopsies from patients with IgA nephropathy. PMID- 8222373 TI - Determination of inulin clearance by using a single-sample method. PMID- 8222374 TI - Autoimmune renal disease and tumour necrosis factor beta gene polymorphism. AB - The genes encoding tumour necrosis factors (TNF) are located within the major histocompatibility complex. Since TNF may be involved in the pathogenesis of autoimmune disease the purpose of the present study was to investigate TNF beta gene polymorphism in two types of immune complex mediated glomerulonephritis, IgA nephropathy (IgAN) and idiopathic membranous glomerulonephritis (IMN) and to compare them with IDDM and healthy controls. DNA was studied by Southern-blot hybridisation methods using Nco I digestion and a TNF beta probe; two alleles were detected size 5.5 kb and 10.5 kb. In healthy controls (n = 107), 9% were 5.5 homozygotes, 47% heterozygotes and 44% 10.5 homozygotes. The corresponding figures in IMN (n = 51) were 21.5%, 61% and 17.5% (p = 0.002), in IDDM (n = 42) 24%, 50% and 26% (p = 0.027) and in IgAN (n = 77) 2.5%, 65% and 32.5% (p = 0.025). The increase in 5.5 homozygotes in both IMN and IDDM was found to be due to an increased frequency of the haplotype A1-B8-TNF beta 5.5-DR3 seen in both these diseases; whereas in IgAN the increased frequency of the 10.5 kb allele can be explained by an association of a Taq 1DQB1-T2 allele with the TNF beta 10.5 allele. These results demonstrate an association of TNF beta gene polymorphism with IMN and IgAN and confirm the associations found in IDDM. Although these disease associations can be explained by linkage disequilibrium with extended MHC haplotypes, a direct role of genetically determined TNF production in the etiology of these diseases remains to be excluded. PMID- 8222375 TI - Renal failure due to mesangiocapillary glomerulonephritis in pregnancy: use of plasma exchange therapy. AB - A 20-year-old female became pregnant 4 years after diagnosis of type I mesangiocapillary glomerulonephritis. Despite normal serum creatinine at conception renal function deteriorated during pregnancy. The use of plasmapheresis and albumin substitution as well as antihypertensive therapy enabled the continuation of the pregnancy from 27 weeks' gestation until a healthy infant could be delivered at 33 weeks. However, an abrupt decline in function at delivery did not reverse and the patient remains dialysis dependent. We conclude that plasma exchange therapy with albumin substitution may be of benefit in women with mesangiocapillary glomerulonephritis when renal function has deteriorated in pregnancy. Stabilization of renal function can allow continuation of the pregnancy until greater fetal maturity makes the delivery of a healthy infant more likely. Although plasma exchange is an experimental therapy, in our hands it appears safe for the fetus and maternal complications were limited to minor vascular access problems. The best prognostic marker in this case was the severity of the most recent renal biopsy rather than the level of renal function or hypertension at the start of pregnancy. This contrasts with most reported cases of pregnancy and primary glomerular disease where irreversible deterioration of renal function was uncommon when renal function at the start of pregnancy was only mildly impaired and hypertension well controlled. PMID- 8222376 TI - Hyporeninemic hypoaldosteronism associated with multiple myeloma: 11 years of follow-up. AB - Hyporeninemic hypoaldosteronism is an important underlying condition, causing hyperkalemia with hyperchloremic metabolic acidosis, disproportionate to the degree of renal insufficiency present. The principal defect in this syndrome is a reduced level of plasma renin activity, which results in secondary hypoaldosteronism. Diabetes mellitus is usually the primary underlying renal disease, though other causes of renal diseases associated with this syndrome have been described. This case report describes for the first time an elderly patient with multiple myeloma, in remission for more than 11 years, associated with the syndrome of hyporeninemic hypoaldosteronism at the time of diagnosis. The complete resolution of the syndrome after vigorous chemotherapy is an intriguing possibility. PMID- 8222377 TI - Differential regulation of brain and atrial natriuretic peptides in hemodialysis patients. AB - We investigated the effect of volume overload on the plasma concentrations of brain and atrial natriuretic peptides as well as cyclic GMP, using specific radioimmunoassays, in 49 patients with chronic renal failure on regular hemodialysis treatment. Markedly elevated levels of the brain (16.2 +/- 1.3 pmol/l) as well as atrial (39.0 +/- 2.8 pmol/l) natriuretic peptide in plasma were found before the dialysis session, as compared to healthy volunteers (range for brain natriuretic peptide, 0.7-7.3 pmol/l, mean level 2.55 +/- 0.32 (SEM) pmol/l). In contrast to the levels of the atrial natriuretic peptide, those of the brain natriuretic peptide were lowered less efficiently by the dialysis procedure: The mean pre-/postdialytic concentration differences were -1.5 pmol/l and -14.2 pmol/l for brain and atrial natriuretic peptide, respectively. The concentrations of the intracellular mediator of the natriuretic peptides, cyclic GMP, were found to be excessively elevated (34.8 +/- 2.8 nmol/l) and returned to near-normal values (12.4 +/- 1.6 nmol/l) at the end of the dialysis session. Concentrations of BNP in plasma of the patients were well correlated to those of ANP. Significant though less marked correlations were also observed between the plasma concentrations of cyclic GMP and BNP, or ANP, respectively. In contrast to those of ANP, pre-/postdialysis differences in plasma BNP concentrations were not correlated to the extent of volume reduction during dialysis. Our findings show that pathophysiologic states resulting in elevations of the plasma concentrations of the atrial natriuretic peptide can also lead to increased levels of the brain natriuretic peptide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222378 TI - Atrial natriuretic peptide release and volume regulation following kidney transplantation. AB - The major stimulus for atrial natriuretic peptide (ANP) release is atrial stretch and increased values are observed in volume overload states such as chronic renal failure. Since successful kidney transplantation restores volume homeostasis, we compared the effects of human cadaveric kidney transplantation on time course and changes of plasma ANP in the early postoperative period in 4 patients with successful and 4 patients with failed transplantation. ANP concentrations were elevated before transplantation in both groups (91 +/- 16 and 70 +/- 32 pmol/l) and decreased after successful (50 +/- 27 pmol/l, day 16) but increased after failed transplantation (146 +/- 45 pmol/l, day 16). Moreover, there was a close correlation between changes of body weight and ANP concentrations. Plasma renin activity decreased and plasma noradrenaline increased non-significantly in both groups, the latter more so after failed transplantation (116 +/- 42 to 194 +/- 156 vs 156 +/- 157 to 425 +/- 287 ng/l). No correlation was found between changes of renin activity or plasma catecholamines and ANP concentrations. The results indicate that the mechanisms governing release of atrial natriuretic peptide are operative in patients with chronic end-stage renal failure and after successful kidney transplantation with a return of atrial natriuretic peptide concentrations towards normal in the latter. PMID- 8222379 TI - Effect of withdrawal of folic acid supplementation in maintenance hemodialysis patients. AB - The requirement of patients on maintenance hemodialysis for dietary supplements of folic acid is controversial. High levels of folate carry the risk of toxicity as well as being unnecessary. We followed a group of 41 patients, not receiving erythropoietin, for 16 months after the cessation of folate supplementation (5 mg/day). Diet supplied 60-80 g protein and 120-260 micrograms folic acid/day. Red cell folate levels decreased linearly from a mean of 1931 +/- 888 micrograms/l (+/- SD) to 676 +/- 294 micrograms/l after 6 months before levelling off at 455 +/- 222 micrograms/l after 9 months. Mean values were unchanged 7 months later (491 +/- 319 micrograms/l). No patient developed folate deficiency. Hemoglobin values at 6, 9 and 16 months were slightly higher than the baseline value of 8.3 +/- 1.8 g/dl (p < 0.05). Mean corpuscular volumes were generally within normal limits, and vitamin B12 status was satisfactory. We conclude that folic acid supplements are unnecessary in adequately nourished hemodialysis patients who are not receiving erythropoietin. PMID- 8222380 TI - Estrogen therapy for the prevention of coronary heart disease: what are the facts? PMID- 8222381 TI - Recalibration of normal sinus heart rate: clinical thinking and epidemiologic support. PMID- 8222382 TI - Antiplatelet therapy in the treatment of cerebrovascular disease. AB - Aspirin and the new agent ticlopidine have been the most thoroughly evaluated of the platelet-antiaggregating drugs used for the prevention of stroke and other vascular events. Numerous trials have shown aspirin to be effective in reducing the risk of myocardial infarction (MI), recurrent transient ischemic attacks, stroke, and vascular death in men at high risk for these events. Primary prevention trials have shown that aspirin reduces the risk of MI in healthy men over 50 years of age but does not reduce the risk of stroke. Two large, multicenter trials have shown that ticlopidine is effective in reducing the risk of fatal and nonfatal stroke in both men and women. Ticlopidine may also be effective in reducing the risk of recurrent stroke in patients who have had a completed thromboembolic stroke. PMID- 8222383 TI - Coronary anomalies: incidence and importance. AB - Most patients with coronary anomalies are asymptomatic. The knowledge of those variations could be important in regard to invasive catheter treatment or bypass surgery. In a retrospective study, the angiographic findings based on 4,016 patients (1985-1989) were analyzed concerning coronary anomalies and malformations. Of the patients studied, 39 (0.97%) had coronary anomalies, and in 26 of these patients it was an anomalous circumflex branch. In 14 cases, the circumflex branch arose from a separate origin in the left aortic sinus. In 11 patients the origin was from the proximal segment of the right coronary artery. A singular coronary artery was found in five patients, originating from the right aortic sinus in two patients and from the left aortic sinus in three patients. An origin of the left coronary artery from the pulmonary artery, a coronary fistula, or an origin of the left anterior descending coronary artery from the RCA could be found in only one patient. Unexpected findings during invasive procedures would suggest a possibly existing coronary anomaly, especially when main branches cannot be opacified by selective contrast medium injection. PMID- 8222384 TI - SIN-1 has no direct myocardial anti-ischemic action. AB - Anti-ischemic drugs may develop their cardiac activity via peripheral (reduction in preload and/or afterload) or cardiac (coronary vasculature, myocardial cell metabolism) effects. The aim of the study was to investigate whether SIN-1, the active metabolite of molsidomine, develops a direct myocardial anti-ischemic property. Three groups of seven patients each were treated with 0.4 mg SIN-1 administered via either the intracoronary (IC) or intravenous (IV) route, or with placebo in a double-blind randomized investigation. SIN-1 had no influence on either the ischemic parameters in the surface electrocardiogram (ECG) or the intracoronary ECG. There was also no change in peripheral or central hemodynamics or in the severity of angina following this low IC or IV dosage. There is no evidence of a direct myocardial anti-ischemic response of SIN-1. The well known anti-ischemic activity of SIN-1 or molsidomine has to be attributed to the proven peripheral and cardiac vascular responses. PMID- 8222385 TI - Abnormal signal-averaged electrocardiogram in patients with Duchenne muscular dystrophy: comparison of time and frequency domain analyses from the signal averaged electrocardiogram. AB - The clinical significance of an abnormal signal-averaged electrocardiogram (SA ECG) in time- and frequency-domain analyses was assessed in patients with Duchenne muscular dystrophy (DMD). Twenty-four DMD patients and 20 age-matched healthy volunteers were studied. The SA-ECG was recorded by time-domain signal processing using the vector-magnitude method and frequency-domain signal processing using the fast-Fourier transform. Abnormal SA-ECGs were based on comparison with controls and eight abnormal SA-ECGs were detected among 24 DMD patients, seven by frequency-domain analysis and four by time-domain analysis. The end-diastolic left ventricular volume was larger in the patients with abnormal SA-ECG than in those with normal SA-ECG (136 +/- 32 ml vs. 77 +/- 43 ml, p < 0.01). Compared with patients with normal SA-ECGs, the DMD patients with abnormal SA-ECGs had a lower left ventricular (LV) ejection fraction (54 +/- 6 vs. 61 +/- 8%, p < 0.05), as an index of LV contractility, and less LV posterior wall velocity (46 +/- 9 mm/min vs. 62 +/- 16 mm/min, p < 0.01), as an index of LV relaxation. The sensitivity and specificity of frequency-domain analysis for predicting malignant ventricular arrhythmias was 60 and 85%, respectively, compared with 30 and 92%, respectively, for time-domain analysis. The combination of time- and frequency-domain analyses can facilitate identification of DMD patients with ventricular tachyarrhythmias associated with depression of cardiac performance. PMID- 8222386 TI - Echocardiographic study of left atrial thrombi in mitral stenosis. AB - Sixty-nine patients with predominant mitral stenosis were examined by echocardiographic means to detect the presence of left atrial thrombi. Forty-nine of these patients were in sinus rhythm and twenty in atrial fibrillation. Four percent of patients in the sinus rhythm group and 45% of those in the atrial fibrillation group had left atrial thrombi. The two risk factors identified for left atrial thrombi in mitral stenosis were atrial fibrillation and left atrial enlargement. PMID- 8222387 TI - A clinical trial evaluating the 24-hour effects of bisoprolol/hydrochlorothiazide 5 mg/6.25 mg combination in patients with mild to moderate hypertension. AB - This study used 24-h ambulatory blood pressure (BP) monitoring to investigate the effectiveness of a novel low-dose combination of bisoprolol/hydrochlorothiazide in adult patients with mild to moderate essential hypertension. Thirty-six patients with stable mild to moderate hypertension (sitting diastolic BP 95-114 mmHg) after a placebo run-in phase received oral bisoprolol/hydrochlorothiazide 5 mg/6.25 mg once daily for 4 weeks in a single-blind regimen. At office visits, BP and pulse were measured with statistically significant reductions (p < 0.01) recorded after 2 and 4 weeks of treatment. Twenty-four-h ambulatory BP monitoring at the completion of therapy revealed significant reductions (p < 0.01) in both systolic and diastolic 24-h, daytime, and nighttime BP, compared with the end of the placebo treatment phase. Systolic and diastolic load were also reduced (p < 0.01). The combination was well tolerated, and overall quality-of-life questionnaire scores indicated an improvement after bisoprolol/hydrochlorothiazide therapy (p = 0.02). No clinically significant changes from baseline in laboratory parameters were observed; in particular, serum potassium was unchanged. This is the first study to demonstrate the 24-h effectiveness of the bisoprolol/hydrochlorothiazide 5 mg/6.25 mg combination, using 24-h ambulatory BP monitoring. In addition, antihypertensive therapy with low doses of bisoprolol/hydrochlorothiazide in combination may improve tolerability. PMID- 8222388 TI - Use of a latex cover sheath for transesophageal echocardiography (TEE) instead of regular disinfection of the echoscope? AB - Better visualization of cardiac structures is an advantage of transesophageal echocardiography (TEE). Because of this, the transesophageal approach is an essential window in the assessment of various cardiovascular disorders. Quick consecutive access to this instrument should not be delayed by time-consuming cleansing procedures of the probe. In this study we used a sterile latex condom in routine TEE examinations as a barrier to contamination of the echoscope, observing practical use and the incidence of perforations. Defects were detected after the procedure by visual inspection and by an airtightness test. Maneuvers or situations associated with an increased likelihood of injury were checked for. In 180 studies a total of 168 patients were consecutively examined; insertion was feasible in all but one case (99.5%). We found 8 of 181 (4.4%) sheaths defective following the procedure. Visual inspection was less sensitive (only 3 of 8) for finding defects than an airtightness test (8 times). Teeth in situ and/or a bite guard are the only sharp edges during insertion and removing the probe; these seem to be the main risk factors in latex perforation. Application of a cover sheath for each examination saves approximately two-thirds of the time otherwise necessary for a cleansing bath, and easy handling of cover sheaths for TEE make them an alternative to regular disinfection in general. For reasons of safety we recommend checking each used cover sheath after examination for holes so as not to jeopardize the following patient by the possible spread of germs from contaminated probes. Not only visual inspection but also a second method, for example, an airtightness test, is mandatory. PMID- 8222389 TI - Traumatic rupture of an aortic ulcerative atherosclerotic plaque producing aortic dissection: a complication of interscapular back blows used to dislodge objects from the esophagus. AB - Penetrating atherosclerotic ulcer of the aorta is a rare entity which usually occurs in the descending thoracic aorta. Herein, we report an unusual case of penetrating aortic ulcer which ruptured into the mediastinum. Interscapular back blows were performed on our patient in an attempt to dislodge an aspirin which she thought was lodged in her esophagus. Unlike previously reported cases of this entity, the penetrating aortic ulcer in our patient was located in the distal thoracic ascending aorta. Diagnosis of penetrating aortic ulcer can be made by utilizing aortography, contiguous dynamic contrast-enhanced computed tomography or magnetic resonance imaging. Treatment consists of adjunctive medical therapy until surgery can be performed. PMID- 8222390 TI - An integrated software system for quality assurance-related kappa coefficient analysis of intraoperative transesophageal echocardiography interpretive skills. AB - This report describes the development of a quality assurance-oriented integrated software system designed for an anesthesiology-based intraoperative transesophageal echocardiography service. Entry data include patient and operation demographics, two-dimensional echocardiographic, saline-contrast, and color flow/pulsed Doppler assessments of the heart and great vessels, presented in a defined sequence. A statistical analysis component (kappa coefficient analysis) allows for comparison of intraoperative real-time interpretations with laboratory interpretations made by experienced full-time echocardiographers on a field-by-field basis. This provides a means of quantifying expertise in each individual aspect of the patient examination sequence. We believe that such self appraisal data are essential for delineating the status and tracking the progress of service being provided. PMID- 8222391 TI - Successful delivery following myocardial ischemia during the second trimester of pregnancy. AB - Acute myocardial infarction during pregnancy is considered to be associated with approximately 50% mortality of both mother and fetus. However, there are not enough data regarding the role of acute myocardial ischemia. We present a 36-year old, pregnant, white female who was admitted twice at 18 and 20 weeks of gestation with acute myocardial ischemia. Cardiac catheterization revealed 70-80% stenosis of the mid left anterior descending artery (LAD) with normal antegrade flow and very good retrograde filling of the LAD from distal collaterals of the right coronary artery. Therefore, due to angiographic suggestion of protected LAD territory, we recommended medical therapy and scheduled a vaginal delivery that was successfully completed without cardiovascular complications. A stress thallium test performed 6 months later was normal, supporting our clinical judgment. In conclusion, every case of a pregnant woman with coronary insufficiency should be treated according to individual coronary anatomy and blood supply to the territory of the diseased artery, and should not be based on the old data in the literature. The decision for revascularization prior to delivery versus medical therapy, or Caesarean section versus natural delivery, should be made by a team of a cardiologist and an obstetrician. PMID- 8222392 TI - Richard Lower. PMID- 8222393 TI - Control of metastatic properties of BL6 melanoma cells by H-2Kb gene: immunological and nonimmunological mechanisms. AB - The effect of class I H-2 antigen expression on the metastatic properties of BL6 melanoma cells was investigated. The BL6-8 clone isolated from the highly metastatic BL6 melanoma did not express H-2Kb gene. Following transfection with the H-2Kb gene, BL6-8 cells displayed a low metastatic potential in the immunocompetent as well as immunosuppressed (X-irradiated) or triple immunodeficient mice with impaired T, B and natural killer (NK) cells function. The expression of H-2Kb gene and the low metastatic ability of transfected BL6 melanoma cells were associated with appearance of cell membrane soybean agglutinin (SBA) and Griffonia simplicifolia 1B4 (GS1B4) lectin-binding carbohydrates. These alterations in cell surface carbohydrates were found to be a result of reduction in sialylation of SBA binding sites and upregulation of the alpha 1.3 galactosyltransferase (alpha 1.3GT) gene. To assess the importance of H 2Kb-induced alterations in cell surface carbohydrates for metastasis formation, BL6-8 melanoma cells were transfected with H-2Kb gene without neor gene cotransfection and selected for adherence to SBA-lectin-conjugated agarose beads. The transfected clones that expressed SBA and GS1B4 lectin-binding carbohydrates were low metastatic. Further analysis of these clones showed that presence of SBA and GS1B4 lectin-binding carbohydrates rather than expression of H-2Kb molecules per se might be responsible for low metastatic potentials of H-2Kb-transfected cells in the immunocompromised mice. Studies of the possible mechanisms responsible for low metastatic ability of H-2Kb-transfected melanoma cells revealed that these cells displayed a reduced ability to adhere to murine pulmonary endothelial cells as well as to laminin and collagen IV. We hypothesized that the observed nonimmunological effects of H-2Kb gene in BL6 melanoma cells is a result of an interaction between the H-2Kb gene and B16 melanoma-specific ecotropic retrovirus. It results in inhibition of this retrovirus production with consecutive alteration in the expression of cellular genes controlling cell surface glycosylation and adhesion properties essential for the metastatic phenotype of BL6 melanoma. PMID- 8222394 TI - Accumulation of heparan sulfate in the culture of human melanoma cells with different metastatic ability. AB - Glycosaminoglycans were metabolically labeled in subconfluent cultures of highly metastatic 7Gp122 and poorly metastatic IC8 variants and of the low metastatic parental M4Be human melanoma cell line. Proteoglycans were separated by DEAE Trisacryl chromatography from the culture medium, from the heparin extract of the cell layer and from the heparin-extracted cell residue lyzed with detergents. Glycosaminoglycans were released from the proteoglycans by reductive alkaline hydrolysis and heparan sulfate (HS) was detected by deaminative cleavage with nitrous acid. Expressed on cell protein basis, the labeled HS content in the medium and in the cell layer decreased with increasing metastatic ability. The extraction of HS with heparin from the 7Gp122 cells indicated that this variant was enriched in (polypeptide bound) HS non inserted into the plasma membrane, compared with the low metastatic IC8 and M4Be cells. The HS fraction in heparin extract and in the heparin-extracted cell residue exhibited molecular mass heterogeneity on gel permeation chromatography and it contained HS fragments. Scission with nitrous acid followed by molecular sieve chromatography of the degradation products indicated that the tetra- and disaccharide repeats separated by the N-sulfated glucosamine residue were present in about equal amounts and constituted 60% of the HS chains in the IC8 and M4Be cells. HS from 7Gp122, IC8 and M4Be cells did not bind antithrombin III with high affinity but it was capable of binding bFGF in in vitro assay. PMID- 8222395 TI - Selection for enhanced adhesion to microvessel endothelial cells or resistance to interferon-gamma modulates the metastatic potential of murine RAW117 large-cell lymphoma cells. AB - Poorly liver metastatic large-cell lymphoma RAW117-P cells were sequentially selected in vitro for increased adhesion to murine hepatic sinusoidal endothelial cells. After three or four sequential selections, the selected sublines showed increased rates of adhesion to target hepatic microvessel endothelial cells and increased formation of experimental metastases in the liver. However, the endothelial cell adhesion-selected RAW117 sublines were generally unstable and gradually lost their enhanced adhesive and metastatic properties during passage in culture. Highly metastatic, liver-selected RAW117-H10 large-cell lymphoma cells were more resistant to the cytostatic effects of interferon-gamma (IFN gamma) than poorly metastatic unselected parental RAW117-P cells. When tested for their sensitivity to IFN-gamma, the endothelial cell adhesion variants were significantly more resistant than the unselected RAW117-P cells, but after a 72-h treatment with IFN-gamma, the in vitro-selected cells lost their enhanced endothelial cell adhesion characteristics, their potential to colonize the liver, and their ability to grow when injected at subcutaneous or intramuscular sites. In contrast, the metastatic potential of similarly treated RAW117-P cells was unaffected by IFN-gamma during a 72-h treatment. Sequential selection of RAW117-P cells for increased resistance to IFN-gamma in vitro resulted in variant lines that were refractory to the growth-inhibiting effects of IFN-gamma, and these IFN gamma-selected variants were also less adhesive to liver microvessel endothelial cells. The IFN-gamma-selected variants also lost their experimental metastatic potentials completely and their tumorigenicities at sites of subcutaneous or intramuscular injection. Cytofluorographic analysis indicated reduced cell surface expression of H-2Kd antigen and fibronectin receptor on the selected variant cells but no change in cell surface mu heavy chain immunoglobulin. The unselected and selected RAW117 lines had similar sensitivities to natural killer (NK) cell-mediated cytolysis, indicating that the in vivo differences were probably not due to differences in NK cell-mediated cytolysis. The results suggest that selection for adhesion to organ microvessel endothelial cells or sequential exposure to certain cytokines can affect the adhesive, growth and metastatic properties of RAW117 cells without modifying their responses to NK cells. PMID- 8222397 TI - Canadian Society for Clinical Investigation and The Royal College of Physicians and Surgeons of Canada Annual Meeting. Vancouver, British Columbia, September 9 13, 1993. Abstracts. PMID- 8222396 TI - Inhibition of tumor metastasis by Arg-Gly-Asp-Ser (RGDS) peptide conjugated with sulfated chitin derivative, SCM-chitin-RGDS. AB - We have synthesized a new compound in which Arg-Gly-Asp-Ser (RGDS) was conjugated with 6-O-sulfated and 6-O-carboxymethyl-chitin (SCM-chitin), i.e. SCM-chitin RGDS, and tested the inhibitory effect on lung and liver metastases of three different types of tumors in mice. SCM-chitin-RGDS was more effective for the inhibition of liver metastasis of L5178Y-ML25 lymphoma and lung metastases of colon 26 M3.1 cells than SCM-chitin, RGDS or their mixture. GRGDS peptide, however, required a higher dose (3000 micrograms) to obtain a sufficiently antimetastatic effect. Intermittent i.v. administration of SCM-chitin-RGDS before or after the i.v. inoculation of L5178Y-ML25 cells caused significant inhibition of liver metastasis as compared with the multiple administration of RGDS, SCM chitin or untreated control. Co-injection of lymphoma cells with SCM-chitin-RGDS or multiple treatment of SCM-chitin-RGDS after tumor inoculation showed significantly enhanced survival rate. SCM-chitin-RGDS also showed the spontaneous lung metastasis produced by intrafootpad injection of B16-BL6 melanoma cells by the multiple i.v. administrations. These results demonstrate that the conjugation of RGDS peptide with SCM-chitin led to augmentation of therapeutic potential to cancer metastasis, thus implying an importance of the conjugation of cell adhesive RGDS peptide with structurally heparin-like SCM-chitin, which possess binding ability to the heparin-binding domain of fibronectin or laminin and extremely low anticoagulant properties. PMID- 8222398 TI - Radiation bone marrow chimeras as a tool to study microglia turnover in normal brain and inflammation. PMID- 8222399 TI - Flow cytometry: measurement of proteolytic and cytotoxic activity of microglia. PMID- 8222400 TI - Neurotoxicity of brain macrophages. PMID- 8222401 TI - Use of rat microglial cells for the study of a possible mechanism of brain damage in AIDS. PMID- 8222402 TI - Microglia: growth factor and mitogen receptors. PMID- 8222403 TI - Microglia, macrophages and the blood-brain barrier. PMID- 8222404 TI - Function-dependent labelling of microglial cells by means of carbocyanine dyes in vivo. PMID- 8222405 TI - Monoclonal antibodies against macrophages/microglia: immunocytochemical studies of early microglial activation in experimental neuropathology. PMID- 8222406 TI - Physiological properties of microglia. PMID- 8222407 TI - Microglial reactions following CNS-grafting. PMID- 8222408 TI - Lessons from microglia in special sites. PMID- 8222409 TI - Long-term results of the Steindler flexorplasty. AB - Seventy-one consecutive patients were treated with a modified Steindler flexorplasty from 1970 to 1987. The mechanisms of injury and etiologies included 45 motor vehicle accidents, 13 birth injuries, and 13 cases of poliomyelitis. Additional operative procedures included shoulder fusion (45 patients), tendon transfer (20 patients), and wrist tenodesis (3). Follow-up averaged 8.2 years. The outcome was excellent in 32%, good in 47%, fair in 13%, and poor in 8%. Postoperatively, the mean arc of active elbow flexion was 114 degrees; the average elbow extension loss, 28 degrees; the mean active pronation, 74 degrees; and supination, 30 degrees. Wire breakage was found in two cases. Additional tendon transfer of flexor carpi ulnaris to extensor carpi radialis brevis improved the outcomes in the patients without active supination. The modified Steindler flexorplasty provided predictable functional improvement in carefully selected patients with paralyzed upper extremities. PMID- 8222410 TI - Brachial artery laceration with closed posterior elbow dislocation in an eight year old. AB - Elbow dislocations are relatively uncommon in children. Rupture of the brachial artery associated with closed elbow dislocations in children is rare. This is a report of an 8-year-old boy, the youngest patient ever to be reported to have a closed posterior dislocation of the elbow associated with a brachial artery laceration. The boy incurred a closed elbow dislocation after a fall onto his outstretched arm. On physical examination, both radial and ulnar (ulnar) pulses were absent. Radiographs showed a posterolateral dislocation of the right elbow and distal fractures of the radius and ulna. Operative exploration of the antecubital fossa showed complete transection of the brachial artery. Repair of the vessel was performed using an interposition vein graft. The distal forearm fractures were managed by closed reduction. At the two-year postoperative follow up examination, the patient had a normal neurovascular examination with full range of motion of his elbow and wrist. Surgical treatment should include exploration of the antecubital fossa and reconstruction of the injured vessels. PMID- 8222411 TI - Allograft salvage of failed total elbow arthroplasty. A report of two cases. AB - Infection after total elbow arthroplasty (TEA) is a devastating complication. Current management options include (1) salvage of the implant with debridement and parenteral antibiotics, (2) resection arthroplasty, and (3) arthrodesis. Most infected TEA patients ultimately require resection arthroplasty. Inadequate bone may preclude both revision TEA and successful resection arthroplasty. It is in these cases that the patient may benefit from allograft reconstruction. Two patients with painful flail elbows secondary to previous resection arthroplasties were treated successfully with cadaver allograft augmented with in situ autograft. Use of the allograft resulted in improvement from failure to excellent in this patient with posttraumatic arthrosis at five years after operation and from failure to good in a rheumatoid patient at two years after operation. Autograft-augmented allografts in the resected failed-TEA patient are a valuable and bone-preserving option. This may be especially useful in patients with significant bone loss or young patients with posttraumatic arthrosis, for whom arthrodesis may be the only other surgical option. The method should not preclude further revision TEA. PMID- 8222412 TI - Complex palmar dislocation of the thumb metacarpophalangeal joint. A case report demonstrating paradoxical function of the extensor pollicis longus. AB - Thumb metacarpophalangeal (MCP) joint dislocations are infrequent injuries, and palmar dislocations rarely occur. The following is a case report of a complex palmar dislocation of the thumb MCP joint that demonstrates the unique physical finding of paradoxical flexion of the MCP joint (and interphalangeal joint extension) in conjunction with the patient's active attempt to extend the MCP joint. This finding proved to be indicative of extensor pollicis tendon displacement palmar to the MCP joint. This soft-tissue incarceration necessitated open reduction. A careful clinical examination is essential for an accurate evaluation of these injuries. Thus, a satisfactory clinical outcome can be attained with strict attention to the intraoperative pathology and technique of repair. PMID- 8222413 TI - Surface anatomy of the hand. The relationships between palmar skin creases and osseous anatomy. AB - Markers placed along palmar skin creases of 53 hands radiographically demonstrated creases superimposed on osseous anatomy. The distal and middle digital creases were found to be consistently proximal to associated interphalangeal joints by 7 to 8 mm and 2 to 3 mm, respectively. Proximal digital creases were consistently distal to associated metacarpophalangeal (MCP) joints by 14 to 20 mm. Distances between the distal transverse palmar crease and associated MCP joints were 6.8 to 10.3 mm. Distances between the proximal palmar crease and the associated MCP joints were 9.1 to 22.1 mm. Mean perpendicular distances between the distal wrist crease and center of selected carpal structures or joints were as follows: scaphoid, 0.7 mm; pisiform, 3.4 mm; lunate, 9.2 mm; hamate hook, 12.6 mm; ulnar styloid, 11.7 mm; trapeziometacarpal joint, 19.4 mm; radioscapholunate joint, 13.5 mm; and distal radioulnar joint, 21.1 mm. Quantification of these relationships should assist in hand examination and placement of surgical incisions and provide further insight into anatomic and functional relationships of the hand. PMID- 8222414 TI - Use of a Herbert screw for interphalangeal joint arthrodesis. AB - Fifty-one Herbert-screw fixation procedures were performed for interphalangeal fusion in the hand. The preoperative diagnoses included rheumatoid arthritis, degenerative arthritis, posttraumatic arthritis, and chronic mallet finger. All patients were observed until there was clinical and radiographic evidence of union. Solid osseous union occurred in all patients. Herbert screw fixation for interphalangeal arthrodesis was a simple, effective technique permitting early mobilization; complications were minimal. PMID- 8222415 TI - Classification of femoral abnormalities in total hip arthroplasty. AB - Bony abnormalities of the femur can significantly complicate total hip arthroplasty both for the primary and revision operations. No standard nomenclature exists for the description of these femoral abnormalities. A classification system is presented to standardize nomenclature, assist in preoperative planning, and to assist in the reporting of these defects. PMID- 8222416 TI - Surgical arthrodesis of the neuropathic foot. A salvage procedure. AB - Reconstructive foot and ankle surgery is a salvage procedure in the deformed neuropathic foot, despite condemnation by some authors. Clinical union and stability was achieved in 91% of the patients, and soft-tissue coverage without skin breakdown was achieved in 100% of the cases. Although one patient had moderate to severe pain in her ankle after operation and was able to do only bed to-wheelchair transfers, she had good clinical stability, no skin ulceration, and was satisfied overall with the procedure. In addition, a significant component of her pain was believed to be from diabetic neuropathy and not pain that was directly attributable to her reconstructive surgery. All other patients were able to ambulate to some degree. More than half had unlimited use of the affected lower extremity. More than half of the patients had mild or no pain, and all patients had a functional limb. Surgical arthrodesis of the deformed neuropathic foot as a salvage procedure can preserve the limb as a stable functional unit, and create an acceptable alignment of the ankle-foot complex that will promote viability of the overlying soft-tissue structures. PMID- 8222417 TI - A 12- to 18-year radiographic follow-up study of Charnley low-friction arthroplasty. The role of the center of rotation. AB - In a 12- to 18-year radiographic follow-up study of 95 Charnley low-friction total hip arthroplasties (THA), there was a statistically significant correlation between location of the center of rotation of the total hip prosthesis (notably the element horizontal distance cup to tear-drop) and long-term, unfavorable, radiographic signs, such as acetabular and stem demarcation, cup wear, cup migration, subsidence of the stem, and calcar resorption. Small differences, as little as 2 mm, in the placement of the cup in relation to anatomic landmarks were responsible for these findings. A logistical regression analysis showed that the horizontal distance cup to teardrop was the most significant parameter influencing long-term unfavorable radiographic signs of a THA in comparison with the parameters of age, gender, diagnosis, and body-weight. PMID- 8222418 TI - Total hip arthroplasty in cerebral palsy. Long-term follow-up results. AB - Patients with cerebral palsy frequently develop coxarthrosis after acquired hip dislocation or dysplasia. Nineteen total hip arthroplasties (THA) were performed in 18 patients with cerebral palsy and end-stage hip degeneration. The average age at arthroplasty was 30 years (range, 16-52 years). All components were cemented, and four hips required bone graft augmentation of the deficient acetabulum. Spica casts were used in 16 of 18 patients to minimize the incidence of dislocation and trochanteric nonunion. The average follow-up time was ten years (range, three to 17 years). Seventeen of 18 patients (94%) had pain relief and improved function after arthroplasty. One stem loosened at three years, and one acetabular component loosened at 15 years. One stem and one acetabular component were revised for malposition. Survivorship analysis was 95% at ten years for loosening and 86% with removal for any reason. Total hip arthroplasty can provide long-term relief and improved function in cerebral palsy patients with severe coxarthrosis. PMID- 8222419 TI - A comparison of the disassociation strength of modular acetabular components. AB - Five short-term in vivo disassembly of two-piece acetabular cup designs have been reported. This study evaluates the liner retention strengths of eight contemporary cup systems. Both push-out (663 +/- 65.5 pounds force to 29 +/- 1.4 pounds force) and lever-out (684 +/- 114 inch-pounds to 43 +/- 1.5 inch-pounds) test modes show a wide variation in retention strength. Repeat liner separation testing demonstrates a 26% and 32% respective decrease in locking mechanism integrity. These findings indicate that reseating modular liners at the time of surgery or reassembling a previously separated liner should be avoided. PMID- 8222420 TI - Evaluation of fractures of the femoral head using the CT-directed pelvic oblique radiograph. AB - Ten patients with femoral head fractures were evaluated with conventional hip radiographs, computed axial tomography (CT), and pelvic oblique radiographs. The CT scan was used to determine the plane of the fracture within the femoral head. The patient was then positioned with the fracture line parallel to the radiographic beam, resulting in a pelvic oblique radiograph. Intraoperative findings were correlated with the preoperative diagnostic studies. The CT directed pelvic oblique radiograph was found to be the most accurate determinant of the extent of fracture displacement and joint congruency, thereby facilitating the choice of appropriate fracture treatment. Most importantly, similar radiographs, reproducible from one examination to the next, could readily be obtained after operation, providing excellent information concerning the adequacy and maintenance of fracture reduction and the progression of fracture healing. PMID- 8222421 TI - Ipsilateral hip and femoral shaft fractures. AB - Forty-two cases of ipsilateral hip and femoral shaft fractures are reviewed. All patients were treated operatively for both fractures, with the exception of the shaft fracture in one child. Several different methods of fixation were used. The most common method was placement of multiple pins or screws around a previously placed intramedullary rod. Initial rodding of the shaft was not associated with avascular necrosis of the femoral head. Nonunion of the femoral neck occurred in three patients as a result of initial displacement and subsequent malreduction. Delays in operative fixation up to six weeks did not affect the ability to obtain union of the femoral neck. Complications involving the shaft fracture were at least as common as complications involving the femoral neck, suggesting that this component of the double injury should receive equally serious consideration. The hip fracture was initially overlooked in 13 cases (31%) without subsequent nonunion or avascular necrosis. Overall, the prognosis with regard to healing of the femoral neck and avascular necrosis of the femoral head in these combination injuries was found to be superior to that of isolated femoral neck fractures in young adults. PMID- 8222422 TI - Robotic assistance in orthopaedic surgery. A proof of principle using distal femoral arthroplasty. AB - The term "robot" refers to a precision mechanical device that is accurately controlled by a computer using intelligent software. The term "robotic assistance" refers to the use of such a device to aid a surgeon in the optimal conduct of a procedure, particularly one requiring specified geometrical relationships. The authors have been exploring the application of robotic assistance in situations in which accuracy and precision are required in orthopaedic surgery. The initial application concerned the planning, positioning, and orientation cuts and holes of the bone required for the femoral component of a total knee arthroplasty. A three-dimensional digitizing template allowed the surgeon to specify the desired position and orientation of the component's articular surfaces in relation to the distal femur. The robotic system used this spatial relationship, along with its knowledge of the geometry of the component selected by the surgeon, to plan the precise location of the required bone cuts and holes. Finally, the robotic assistant sequentially positioned saw and drill guides with respect to the distal femur so that the surgeon made these cuts and holes in the locations necessary for optimal component fit, position, and orientation. The robotic assistant functioned easily in the operating room environment; increased the accuracy; and decreased the time, equipment, and personnel required for the conduct of the geometrical part of this surgical procedure. PMID- 8222423 TI - Chronic ankle instability as a cause of peroneal tendon injury. AB - Chronic lateral ankle pain and instability can be associated with a split in the peroneus brevis tendon. This case reports a peroneus brevis split that was centered over the sharp posterior edge of the fibula and was associated with laxity of the superior peroneal retinaculum, and chronic ankle instability. The mechanism of the split was easily demonstrated during surgery by everting the foot and pulling longitudinally on the peroneus longus tendon. The interrelationship of lateral ankle instability with superior retinacular laxity and resultant peroneus brevis splits can account for posttraumatic lateral ankle pain. Surgical treatment must identify and correct the underlying pathology and should attempt to repair or debride the peroneus brevis tendon, reconstruct the superior peroneal retinaculum, flatten the posterior edge of the fibula by removing the sharp bony prominence, and address any associated lateral ankle instability with either a modified Chrisman-Snook, Anderson, or modified Brostrom Gould procedure. PMID- 8222424 TI - Comparison of tuberculous and pyogenic spondylitis. An analysis of 122 cases. AB - One hundred twenty-two patients with tuberculous or pyogenic spondylitis were investigated retrospectively. Patient histories, laboratory tests, and radiographic findings were compared statistically between the two groups. Significant differences were calculated for the interval between onset of symptoms and diagnosis, erythrocyte sedimentation rate, mean vertebral loss at discharge, and sclerosis of the vertebral bodies involved. Open or closed biopsy was performed in 91 patients. The result provided a clear distinction between tuberculous and pyogenic spondylitis in 62.2%, either by means of histology or by culture growth. In pyogenic spondylitis, staphylococci were the most predominant bacteria isolated. Neurologic deficits were demonstrated in 17.8% of patients with tuberculous spondylitis and 22.7% with pyogenic spondylitis. At follow-up examinations, only two patients still had a motor deficit. Additionally, pain, gibbus formation, and bony fusion were evaluated, but no significant differences were found. The combination of several unspecific findings such as patient history, erythrocyte sedimentation rate, and radiographic assessment can lead to the correct diagnosis. A definitive diagnosis is established by means of biopsy, histologic evidence, and bacterial culture. PMID- 8222425 TI - Diabetes mellitus. PMID- 8222426 TI - Intramedullary supracondylar nailing of femoral fractures. A preliminary report of the GSH supracondylar nail. AB - From 1987 through 1990, 34 acute supracondylar femoral fractures in 33 patients were treated with an interlocking closed-section intramedullary nail inserted retrograde into the femur through the intercondylar notch. Follow-up data of at least five months were available for 25 of these fractures. The patients suffered injuries more extensive than comparable patients reported in the literature. All fractures healed clinically and radiographically. The average arc motion at the most recent evaluation was 100 degrees. The average operative time was 156 minutes and the average estimated blood loss was 224 ml. Complications related to this treatment were successfully resolved. The genucephalic nail is a good treatment for supracondylar femoral fractures. PMID- 8222427 TI - Treatment of tibial defects and nonunions using ipsilateral vascularized fibular transposition. AB - Eight patients had nine ipsilateral vascularized fibular transpositions (IVFTs) in the period 1978-1987. The procedure, which involves no microvascular anastomosis, was done for difficult problems of the tibia including two long standing posttraumatic nonunions, three allograft nonunions after tumor surgery, and four segmental tibial defects also after tumor surgery. The average duration of problems before surgery was 3.5 years, and the patients averaged 3.1 procedures before IVFT. The patients were followed for an average of 52.4 months. The average time to union was 4.2 months, and in all patients the grafts healed within six months. Late fracture of the graft developed in two patients after fixation removal; one required an additional procedure, and both eventually healed. Ipsilateral vascularized fibular grafting is a useful alternative to conventional, nonvascularized grafts for difficult tibial nonunions and segmental defects. It offers the advantages of a vascularized graft (early healing and hypertrophy), yet avoids the time-consuming microvascular anastomosis and distant donor site morbidity of free fibular grafts. PMID- 8222428 TI - Exostectomy for symptomatic bony prominences in diabetic charcot feet. AB - Exostectomy for plantar ulceration is demonstrated to be a satisfactory procedure and one with significantly less morbidity and quicker healing than a more major reconstructive procedure such as arthrodesis. The predominance of Type I (midfoot breakdown) confirms reports on the natural history of the diabetic Charcot tarsus; the midfoot has a tendency to develop symptomatic bony prominences. PMID- 8222429 TI - Intramuscular metastasis of carcinoma. AB - Four cases of intramuscular metastasis of carcinoma are reported. The preoperative clinical diagnosis of all four cases was soft-tissue sarcoma. Definitive diagnoses, made by open biopsy, were uterine carcinoma, gastric carcinoma, lung carcinoma, and hypopharyngeal carcinoma. All patients died from lung, liver, or brain metastasis several months after open biopsy despite surgery, chemotherapy, or radiation therapy. Case reports of macroscopic metastases to muscle are rare, and the authors report four such cases experienced during a 15-year period. PMID- 8222430 TI - Osteoid osteoma under the age of five years. The difficulty of diagnosis. AB - Ninety-one cases of histologically confirmed osteoid osteoma were collected during an 11-year period; of them, seven (7.6%) had onset of symptoms while younger than five years of age. Special diagnostic difficulties were found in this specific age group: most cases were misdiagnosed or diagnosed incorrectly. The time between the onset of symptoms and the diagnosis varied from three months to five years. Although pain was present in six patients, in four cases, other concomitant signs and symptoms attracted more attention and led to a misdiagnosis and unnecessary invasive procedures. Gait disturbance (limp) was the second most frequent sign and was always present when the lower extremity was affected. In five cases, initial radiographs were not conclusive. Bone scans were very sensitive and conclusive in three cases where radiographs were atypical. Computerized tomograms always located the tumor. PMID- 8222431 TI - Osteosarcoma of the talus. A case report. AB - A 20-year-old man complained of increasing pain and swelling in the right ankle joint. Radiographs of the ankle demonstrated an ill-defined osteolytic lesion in the talus associated with a small, round bone formation just proximal to the neck of that bone. Histologic examination of the lesion showed osteoblastic osteosarcoma originating in the talus. Below-knee amputation was performed because of the peripheral location of the tumor and the close anatomic confines of the foot. A literature review demonstrated that the calcaneus and metatarsals are favorite sites of this tumor in the foot, with only one case in the talus. A second case of osteosarcoma in the talus seems to have not been previously reported. PMID- 8222432 TI - Characterization of a cell line derived from a human giant cell tumor that stimulates osteoclastic bone resorption. AB - Giant cell tumors of bone are common but unusual tumors that are comprised of multiple cell types. Most attention has been focused on the giant cells, which resemble osteoclasts morphologically and functionally. This study examines the properties of a cell line derived from mononuclear cells from one of these tumors, since it appears likely that these cells may be able to influence the activities of cells with the osteoclast phenotype. This cell line, C433, has the following characteristics: (1) it represents undifferentiated cells, not recognized by any known antigenic markers for leukocytes; (2) it contains tartrate-resistant acid phosphatase; (3) it responds to the osteotropic factors 1,25 dihydroxyvitamin D3, insulin-like growth factor I and II, but not to parathyroid hormone; (4) it forms sarcomas in nude mice; and (5) it produces an activity that stimulates isolated avian and rat osteoclasts to resorb bone. This cell line may be useful in examining interactions between osteoclasts and accessory cells involved in bone resorption. PMID- 8222433 TI - Antibiotic impregnated bone cement in total hip arthroplasty. An in vivo comparison of the elution properties of tobramycin and vancomycin. AB - A prospective in vivo quantification was performed to measure the elution of tobramycin and vancomycin antibiotics from two commonly used bone cements. Forty patients were divided into four groups: Group I, tobramycin-Simplex; Group II, tobramycin-Palacos-R; Group III, vancomycin-Simplex; and Group IV, vancomycin Palacos-R. Antibiotic levels were measured from hemovac wound drainage, urine, and serum and compared with control groups who received either intravenous tobramycin or vancomycin. There were no significant differences in daily mean tobramycin levels in hemovac samples between Groups I and II. Tobramycin hemovac levels from Groups I and II were significantly higher than the tobramycin control group. Similarly, no differences were seen in daily mean vancomycin levels of the hemovac samples between Group III and IV; however, the intravenous vancomycin control group had significantly higher levels in the hemovac fluid than Groups III or IV. Tobramycin in the hemovac fluid from Groups I and II was highly bioactive against the control organism. Vancomycin in the hemovac fluid from Groups III and IV had variable bioactivity against the control organism. In 30% of the cases, no vancomycin was detected in the hemovac fluid, and in these cases, the hemovac fluid had no effect on the control organism. Tobramycin elutes to give adequate local tissue levels and releases antibiotic effects when used in an antibiotic bone cement combination. Vancomycin has variable elution properties and is not a predictable additive for the bone cements tested. PMID- 8222434 TI - Use of parenteral prophylactic antibiotics in clean orthopaedic surgery. A review of the literature. AB - Parenteral prophylactic antibiotics are now a routine part of most clean orthopaedic procedures since so many animal and clinical studies have shown a reduced infection rate with their use. First-generation cephalosporins are theoretically the preferred agents, and the pharmacokinetics of Cefazolin make it the drug of choice. It is imperative that an antibiotic be given on the induction of anesthesia or at least ten minutes before inflation of a tourniquet. Based on evidence in the current literature, 24 hours of postoperative coverage appears to be adequate. PMID- 8222435 TI - The pathophysiologic role of fat in dysbaric osteonecrosis. AB - Dysbaric osteonecrosis (DON) can occur in humans and sheep after a single hyperbaric air exposure with inadequate decompression. The authors hypothesize that DON does not result from primary embolic or compressive effects of nitrogen bubbles on the osseous vasculature, but by secondary injury to the marrow adipose tissue by rapidly expanding nitrogen gas that triggers local, and possibly systemic, intravascular coagulation. A 28-year-old scallop diver remained at a depth of 92 feet in sea water for 4.5 hours on surface-supplied compressed air. Decompression sickness occurred after a no-stop ascent to the surface, and he died 70 minutes later. Autopsy showed multiple gas bubbles, not only within the great vessels, but in the fatty marrow of his femoral and humeral heads. Lipid and platelet aggregates were found on the surface of marrow bubbles. Fibrin platelet thrombi were detected within dilated venous sinusoids adjacent to bubbles, and in veins, capillaries, and arterioles. Since pulmonary, renal, and intraosseous (subchondral) fat embolism and fibrin thromboses were observed, it is suggested that injured marrow adipocytes can release liquid fat, thromboplastin, and other vasoactive substances, which conceivably can also play a systemic procoagulant role in triggering disseminated intravascular coagulation and additional DON. PMID- 8222436 TI - Supraventricular tachyarrhythmias during total joint arthroplasty. Incidence and risk. AB - Perioperative supraventricular tachyarrhythmias (AF/SVT) have been long recognized as a complication after major surgery, but little is known about the incidence after major nonthoracic surgery. One thousand two hundred ten consecutive patients undergoing total hip or knee arthroplasty were studied to determine the incidence of new onset AF/SVT. Information on preoperative medical history and laboratory tests were collected in a subset of 583 patients, and analyzed using logistic regression and linear analysis to determine risk. Ninety four-and-one-half percent of patients received an epidural anesthetic, 4.1% had general anesthesia and 1.3% had spinal anesthesia. New onset AF/SVT was found in 38 of 1210 patients, representing an incidence of 3.1%. In the subset of 583 patients, the incidence was 4.8%. The only variables found to be independently associated with the perioperative development of AF/SVT were a history of atrial fibrillation, increasing age, left anterior hemi-block, and atrial premature depolarizations on the preoperative electrocardiogram. In those patients 60 years of age or older with one or more positive risk factors (13% of the study population), the incidence was 18.2%. In those patients less than 60 years of age with none of the identified risks, the incidence was 1.9%. PMID- 8222437 TI - Treatment of the diabetic foot from a vascular surgeon's viewpoint. AB - Diabetic foot lesions are caused by neurologic, orthopaedic, immunologic, and vascular derangements. Whereas some lesions may be initially caused by trauma and others directly caused by vascular disease, improvement of arterial perfusion often plays an important role in the successful salvage of these limbs. Fortunately, in the last several years, there has been a major improvement in the identification and treatment of those patients in whom correction of arterial occlusive disease is necessary for healing. PMID- 8222438 TI - Gait analysis in patients after Van Nes rotationplasty. AB - Gait was analyzed through kinetic, kinematic, and temporal-distance parameters assessment to evaluate the functional performance of ten children treated by the classical Van Nes rotationplasty (VNR). Van Nes rotationplasty is a surgical procedure, which permits the conversion of the ankle joint into a "pseudo-knee" joint. The data were compared with measurements of a normal population of ten healthy subjects. Significant differences with respect to the normal were found in stride duration, stride length, cadence, velocity, and stance-swing ratio, and in ground-reaction forces parameters, which define the propulsive phase in the prosthetic side and the acceptance phase in the sound side. There were differences in some kinematic parameters as well. Van Nes rotationplasty is acceptable from a biomechanical-functional point of view, particularly with respect to the active control of the pseudo-knee, which allows a smooth and coordinate gait pattern. PMID- 8222439 TI - Effect of screw diameter, insertion technique, and bone cement augmentation of pedicular screw fixation strength. AB - This study investigated (1) the effect of screw diameter and insertion technique in lumbar vertebrae, and insertion site in the sacrum, on the axial pullout force and transverse bending stiffness of pedicle screws, and (2) the effect of bone cement augmentation using polymethylmethacrylate (PMMA) and the biodegradable composite, poly(propylene glycol-fumarate) on axial pullout force and transverse bending stiffness of pedicle screws inserted into lumbar vertebrae. The axial pullout force and transverse bending stiffness of a 6.25-mm Steffee screw and a 6 mm Kluger screw did not differ significantly in vertebral bodies of similar equivalent bone mineral density. The axial pullout force of Schanz screws was significantly increased with a 1-mm increase in screw diameter. However, there was no significant increase in transverse bending stiffness. In the sacrum, an approach through the S1 facet produced significantly higher axial pullout forces and transverse bending stiffness than the approach described by Harrington and Dickson. PMMA and a biodegradable composite bone cement poly(propylene glycol fumarate) both increased the axial pullout force. PMMA also increased the transverse bending stiffness. PMID- 8222440 TI - Effects of acute graded strain on efferent conduction properties in the rabbit tibial nerve. AB - This study examines effects of acute stretch on electrophysiologic properties of rabbit tibial nerve. Compound motor action potentials (CMAP) were measured in 30 rabbits, during and after two hours of acute, graded strain. The rabbits were assigned to one of three groups defined by the degree of stretch (0%, 8%, or 15%) beyond original resting length. These groups were subdivided according to ankle position during nerve strain (ankle dorsiflexion or ankle plantar flexion). Two hours of 8% strain did not significantly decrease CMAP amplitude. Fifteen percent strain with the ankle in dorsiflexion produced a significant and profound drop (99%) in CMAP amplitude. These findings are clinically relevant to nerve stretch injuries and to surgical procedures that induce acute nerve strain. PMID- 8222441 TI - Bone morphogenetic protein-induced muscle- and synovium-derived cartilage differentiation in vitro. AB - Muscle- and synovium-derived mesenchymal-type cells differentiated into cartilage in response to a coating of partially purified rabbit bone morphogenetic protein (BMP) on a substratum of plastic. Observed by means of phase contrast microscopy, light microscope histochemistry, and electron microscopy, the sequence of events on a BMP-coated plastic substratum is the same as that observed on a substratum of bone matrix. The plastic eliminates the possibility of any endogenous allogeneic matrix-derived BMP contaminating the system. The differentiation of cartilage from synoviocytes and subsynovial cells resembles the pathologic process occurring in human chondromatosis. PMID- 8222442 TI - Effects of constant mechanical tension on the healing of rabbit flexor tendons. AB - The biomechanical effects of constant mechanical load on tendon repair in vitro were determined for rabbit flexor tendons. Tendons were removed from Zone II, transected, reapproximated with four simple sutures, and cultured in standard medium. Tendons from the right forelimbs were loaded with 3.1-g weights; tendons from the contralateral forelimbs served as unloaded tendons. Tenorrhaphies were disrupted at zero, one, three, and six weeks postsuturing by fixed-speed tensiometry. True maximum stress (strength), normalized energy absorbed, and tangent modulus steadily increased over time, becoming significantly greater than unincubated controls in the loaded and unloaded groups at six weeks. True strain at maximum stress increased with duration for unloaded tendons; after six weeks it was significantly greater than unincubated control tendons. This study demonstrates a method for quantifying the biomechanics of tendon after intrinsic tendon segment healing and presents the first biomechanical evaluation of constant tension applied across the laceration site during an in vitro healing phase. PMID- 8222443 TI - A case of bilateral dysplasia epiphysealis hemimelica associated with polydactyly and syndactyly. AB - Dysplasia epiphysealis hemimelica (DEH) on bilateral medial malleoli occurred in a boy who had polydactylies and syndactylies of all four limbs. Cases with both bilateral and symmetrical DEH affection as in this case seem not to have been reported previously in the literature. Dysplasia epiphysealis hemimelica complicated by congenital anomalies is extremely rare. A one-month-old boy of normal delivery had polydactylies of thumbs, small fingers, and great toes, and had symmetrical syndactylies of fingers and toes. At age eight, the patient reported swelling and tenderness on bilateral medial malleoli. Radiographs showed small blotches of radiopacity. Two years later, the radiopacities had enlarged to become typical of DEH. PMID- 8222444 TI - Recent advances in diabetes care and management. AB - Four developments in diabetes care and management have evolved over the last decade and are now routinely incorporated into clinical practice: (1) the introduction of recombinant human insulin; (2) the use of second generation oral agents; (3) home glucose monitoring and glycohemoglobin monitoring; and (4) a better understanding of the relationship between diabetic control and complications. PMID- 8222445 TI - What to do about a dropped bone graft. AB - Autogenic bone graft operations are common. Although it is not often mentioned, most orthopedic surgeons are aware of instances in which the graft has been dropped on the operating room floor. The current forms of sterilization cause damage to the graft and loss of its advantages over allograft bone. To determine the amount of contamination incurred when the graft is dropped, surplus bone specimens from 50 orthopedic and neurosurgical procedures were dropped on the operating room floor and submitted for culture. Fifty controls were obtained from the same cases. No positive cultures were obtained from either study group. These findings offer objective evidence that extensive sterilization of a dropped graft is not essential. PMID- 8222446 TI - Healing of foot ulcers in immunosuppressed renal transplant patients. AB - Sixteen renal transplant patients were retrospectively studied to determine the time foot ulcerations healed. All patients were taking imuran and medrol. More recently, patients were also on cyclosporine. All were insulin-dependent diabetics. A control group was established from a pool of randomly selected, age matched, insulin-dependent diabetics who were also observed for foot ulcerations. All treatment was given by one physician. Pulse volume recordings were obtained in all but eight patients, who were clinically believed to have normal vascular supply to the affected extremity. No statistically significant difference was found between these groups in this regard. Cultures were obtained in all patients when indicated; appropriate antibiotics were started. No major differences were noted with respect to infecting organism. The immunosuppressed group had no excellent results, and three good, 14 fair, and 23 poor results. The control group had ten excellent, 14 good, 11 fair, and 17 poor results. A higher failure rate may occur with limb-salvage surgery in the immunosuppressed patient. This observation confirms a long-held clinical suspicion. PMID- 8222447 TI - On arthropathies of cerebral or spinal origin. PMID- 8222448 TI - A short history of neuropathic arthropathy. AB - Neuropathic joint disease or Charcot joints are a chronic form of a degenerative arthropathy that is associated with decreased sensory innervation of the involved joints. Neuropathic arthropathies are considered to be an accelerated osteoarthritis that is precipitated by trauma to a joint not protected by its proprioceptive or nociceptive reflexes. This process continues until destruction of the joint occurs. The etiology of Charcot joints has varied throughout history as different diseases have become less or more prominent. Some of the offenders include diabetic neuropathy, tabes dorsalis, syringomyelia, and congenital indifference to pain. The morphology and pathogenesis of this condition has intrigued the minds of many, including Musgrave, Charcot, Virchow, Eloesser, Soto Hall, and Key. PMID- 8222449 TI - Osteomyelitis in the foot and ankle associated with diabetes mellitus. AB - Osteomyelitis in a foot or ankle is a common complication in patients with diabetes mellitus. Although acute osteomyelitis can be a surgical emergency, this presentation is uncommon. Chronic bone infection in most patients is associated with either vascular impairment or neuropathic skin breakdown or both. Neuropathic joints and stress fractures have to be differentiated from radiographic abnormalities caused by infection. PMID- 8222450 TI - Management of neuropathic arthropathy with the Charcot Restraint Orthotic Walker. AB - A recently designed Charcot Restraint Orthotic Walker (CROW) was used in the treatment of 18 patients with diabetic neuroarthropathy involving the foot and ankle. Eight of these patients had no surgery before the use of the CROW. In ten patients, the device was used for prolonged immobilization after surgery for complications of neuropathic joint disease. The CROW is a rigid, custom, full foot enclosure ankle-foot orthosis. It was used after an initial period of cast immobilization. The CROW effectively controls limb edema, returns the patient to ambulatory activities, and prevents significant progression of deformity. All patients rated their satisfaction with the device as good or excellent. The CROW is an attractive alternative to currently used methods to provide the prolonged immobilization and protection necessary for healing in neuropathic arthropathy. PMID- 8222451 TI - Amputations in the diabetic foot and ankle. AB - Scientific structured foot salvage clinics will provide surgeons with a large population of peripheral vascular insufficiency patients who may someday become candidates for salvage amputation at the foot or ankle level. This article presents the technology of functional amputation levels. PMID- 8222452 TI - Amputation level selection in the diabetic foot. AB - Walking independence in the diabetic patient with gangrene or infection can be maintained with a conservative program of distal amputation. Patients are preevaluated for their rehabilitation potential. The biologic amputation level (the most distal functional amputation level with a reasonable potential to heal) is determined by a measure of vascular inflow, tissue nutrition, and immunocompetence. By combining wound-healing potential with rehabilitation potential, one can determine the amputation level that will most likely optimize function. PMID- 8222453 TI - Salvage of the diabetic foot with exposed os calcis. AB - Fifty consecutive heel ulcers were managed in three groups by debridement, split thickness skin graft (STSG), bypass procedures, and orthotics. Group I consisted of 24 ulcers in patients with diabetes (DM) and peripheral vascular disease (PVD), 14 patients in Group II with DM only, and 12 patients with PVD only (Group III). Healing occurred in 56.5%, 64.3%, and 83.3%, respectively. An average of 2.2 procedures were performed per patient. Follow-up periods were for a minimum of two years or until amputation. Time for complete healing and the number of amputations performed were similar in all groups. Of the diabetics (combined from Groups I and II), a subgroup of 27% required partial excision of the os calcis to facilitate closure. After saline dressing changes, STSG was accomplished over thin granulation tissue. Forty percent of this subgroup healed, 30% remained open, and 30% were amputated. Aggressive management, soft-tissue coverage, and orthotic use can lead to a functional weight-bearing extremity. PMID- 8222454 TI - Diabetic neuropathy. AB - The neuropathy associated with diabetes mellitus is arguably the most common peripheral neuropathy in the developed world. It can be classified into symmetrical and asymmetrical forms. The symmetrical form is predominantly sensory and autonomic, whereas the asymmetrical form can be sensory, or motor, or both, and can affect individual cranial or peripheral nerves. Pathologic and electrophysiologic studies indicate that the symmetrical polyneuropathy is characterized by a distally accentuated loss of myelinated and unmyelinated axons. The multifocal nature of the axonal degeneration and its association with vascular disease in the same pathologic specimens strongly suggest an ischemic cause involving the endoneurial microvascular circulation. The underlying biochemical abnormality appears to be the production of advanced glycosylated end products (AGEPs), in the presence of chronic hyperglycemia, which accumulate on endothelial proteins, causing basement membrane thickening and endothelial cell change. This produces multifocal vascular disease that, in turn, reduces nerve blood flow, causing endoneurial hypoxia and generating oxygen free radicals. Multifocal ischemic neuronal damage results. Currently, there is no accepted treatment, although long-term control of hyperglycemia is beneficial. Symptomatic relief of the burning pain is the most vexing therapeutic problem; tricyclic antidepressants and anticonvulsants have been used alone and together with varying success. PMID- 8222455 TI - Neuropathic calcaneal tuberosity avulsion fractures. AB - Avulsion of a calcaneal tuberosity can be the initial presentation of a neuropathic fracture. Neuropathic fracture should be investigated in patients who appear without a history of significant indirect or direct trauma. Neuropathic fractures should always be considered in patients with diabetes mellitus. The diagnosis of neuropathic calcaneal tuberosity avulsion fracture alters the prognosis and treatment of this injury. Two cases of bilateral fracture avulsions are reported to emphasize problems in diagnosis and treatment. PMID- 8222456 TI - Chopart amputation with percutaneous heel cord lengthening. AB - When amputation becomes necessary in a patient with peripheral vascular disease, it is important to preserve as much tissue as possible to preserve maximum function. This is especially important because the other extremity may have similar involvement in the future. With appropriate care, an amputation at the Chopart (calcaneocuboid-talonavicular) level can give a good functional result. PMID- 8222457 TI - The posterior deltoid-splitting approach to the shoulder. AB - Since 1980, the authors have used a posterior approach to the glenohumeral joint in which the posterior deltoid is split caudally in line with its fibers from the posterior acromion to the upper border of the teres minor. The extent of this deltoid splitting approach exceeds that of a similar anterior approach because of the distal emergence of the axillary nerve from the quadrilateral space. This technique gives complete access to the infraspinatus and teres minor muscles and tendons, posterior capsule, and posterior glenoid. Unlike traditional posterior approaches to the shoulder joint that detach a portion or all of the origin of the deltoid, this technique preserves the deltoid origin from the scapular spine and posterior acromion. Over the past 11 years, this posterior approach has been performed in 35 patients (42 shoulders): 31 for posterior instability, one for posterior glenohumeral fracture-dislocation, eight for infection, and two for removal of foreign bodies. The median age of the patients was 33.8 years (range, 13-65 years). The mean duration of follow-up contact was 20 months (range, one month to 11.4 years). Two patients died of unrelated causes and three were lost to follow-up examination. The posterior deltoid-splitting approach is advocated for any procedure requiring posterior access to the glenohumeral joint because it provides excellent exposure, has been associated with no complications, and preserves the strength and function of the posterior deltoid. PMID- 8222458 TI - The influence of scapular retraction and protraction on the width of the subacromial space. An MRI study. AB - The width and configuration of the subacromial space was studied in retraction and protraction of the shoulder girdle in four healthy subjects by magnetic resonance imaging. The anterior opening of the subacromial space narrowed as the shoulder moved from a retracted to a protracted position. PMID- 8222459 TI - Cefetamet pivoxil clinical pharmacokinetics. AB - Cefetamet pivoxil is an orally absorbed prodrug ester of the microbiologically active cephalosporin, cefetamet. The prodrug ester is completely hydrolysed to the active compound cefetamet on its first pass through the gut wall, the liver or both. Cefetamet is classified as a third generation cephalosporin with excellent activity against streptococci, Enterobacteriaceae, Neisseria and Haemophilus species. It has enhanced stability against beta-lactamases compared with penicillins and first and second generation cephalosporins. The antibacterial spectrum is comparable with that of cefotaxime except for its poor activity against staphylococci. Following a 20-minute zero-order intravenous infusion, cefetamet had a rapid distribution phase followed by a monoexponential decline. The average pharmacokinetic parameters from 152 healthy volunteers were: total body clearance 136 ml/min (8.16 L/h); renal clearance 119 ml/min (7.14 L/h); nonrenal clearance 17 ml/min (1.02 L/h); volume of distribution at steady state 0.29 L/kg; terminal elimination half-life 2.2 hours; 88% of the dose recovered in the urine. Cefetamet is not extensively bound to plasma proteins. Consequently, these data indicate that cefetamet is predominantly eliminated unchanged by the kidney via glomerular filtration with possibly a minor component of tubular secretion. Cefetamet has a relatively small apparent volume of distribution consistent with that of other beta-lactam antibiotics. Results following ascending intravenous doses of cefetamet in healthy young male volunteers demonstrated that the pharmacokinetics of intravenous cefetamet are independent of the dose. The absolute bioavailability of cefetamet tablets following oral cefetamet pivoxil administration is enhanced by the presence of food. Under fed conditions, 50 to 60% of the final oral dose is absorbed into the systemic circulation. This food effect is observed when cefetamet pivoxil is administered within 1 hour of a meal. Food also produces a slight delay in the time to reach peak plasma concentrations of this drug. Changes in fluid volume intake with cefetamet pivoxil administration have no effect on the bioavailability of this drug. Similar absorption characteristics have been observed for all of the tablet dosage formulations studied during clinical development. The absolute bioavailability of the final syrup dosage formulation was between 38 and 47%. Little improvement in the bioavailability of this preparation has been observed with food. The absorption and disposition of cefetamet in human subpopulations [i.e. children, elderly (< 75 years of age), renal impairment, liver disease and patients taking concomitant drugs] have been studied extensively. Only impaired renal function appears to significantly alter the elimination of this drug.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8222461 TI - Computers in pharmacokinetics. Choosing software for clinical decision making. AB - Over the past 20 years, pharmacokinetic programs have been developed for clinical decision making. These clinical pharmacokinetic software programs are designed to assist the clinician in the analysis, interpretation and reporting of serum drug concentration data for a variety of medications. The programs vary in the extent of features and range of medications supported and thus warrant careful review before selecting or purchasing such a program for routine use. A series of programs which are commercially available in the United States was reviewed for this article. The focus of the review is not to recommend a single program or to provide a ranked list of commercially available programs. Information is presented to clinicians to better their understanding of the features of these computer-based clinical resources. As an introduction to this topic, the information presented concentrates on the system and support features. Those programs that were reviewed demonstrate the ability to assist in the analysis of serum or plasma drug concentration data for most of the medications that warrant therapeutic drug monitoring. They provide both Bayesian and non-Bayesian methods for predicting serum drug concentrations. Standard personal computers were sufficient to run each of the programs reviewed. In addition, most programs offered technical and clinical support. However, the quality of the user manuals and training material varies among software programs. In-depth analytical comparisons are currently being conducted for future publication. PMID- 8222460 TI - Clarithromycin clinical pharmacokinetics. AB - Clarithromycin is a semisynthetic macrolide antibiotic, structurally related to erythromycin. It has a more favourable pharmacokinetic profile than erythromycin, thus allowing twice-daily administration and possibly increasing compliance among outpatients. Clarithromycin is well absorbed from the gastrointestinal tract and its systemic bioavailability (about 55%) is reduced because of first-pass metabolism. It undergoes rapid biodegradation to produce the microbiologically active 14-hydroxy-(R)-metabolite. The maximum serum concentrations of clarithromycin and its 14-hydroxy metabolite, following single oral doses, are dose proportional and appear within 3 hours. With multiple doses, steady-state concentrations are attained after 5 doses and the maximal serum concentrations of clarithromycin and of the 14-hydroxy derivative appear within 2 hours after the last dose. Clarithromycin is well distributed throughout the body and achieves higher concentrations in tissues than in the blood. Also, the 14-hydroxy metabolite exhibits high tissue concentrations, with values about one-third of the parent compound concentrations. The presence of food appears to have no clinically significant effect on clarithromycin pharmacokinetics. The main metabolic pathways are oxidative N-demethylation and hydroxylation, which are saturable and result in nonlinear pharmacokinetics. The primary metabolite (14 hydroxy derivative) is mainly excreted in the urine with the parent compound. A reduction in urinary clearance in the elderly and in patients with renal impairment is associated with an increase in area under the plasma concentration time curve, peak plasma concentrations and elimination half-life. Mild hepatic impairment does not significantly modify clarithromycin pharmacokinetics. In conclusion, clarithromycin, because of its antibacterial activity and pharmacokinetic properties, appears to be a useful alternative to other macrolides in the treatment of community acquired infections. PMID- 8222462 TI - Effects of time of administration and posture on the pharmacokinetics of cefprozil. AB - The effects of time of administration, sleep and posture on the pharmacokinetics of cefprozil were evaluated in a single-dose 3-way crossover study. After a 6 hour fast, 12 healthy male volunteers received oral cefprozil 250mg at 1200h (treatment A), 1200h (treatment B) and 2400h (treatment C) with a 7-day washout interval between each treatment. During the study period, volunteers receiving treatment A remained in a sitting/standing position or were ambulatory, those receiving treatment B were in the supine position, and those receiving treatment C were sleeping. Blood samples were taken over an 8-hour period and the plasma samples were analysed for the concentrations of cefprozil by a high performance liquid chromatography-ultraviolet method. Plasma concentration vs time data were analysed using noncompartmental analysis methods. Mean peak plasma concentrations (Cmax) were 4.51, 5.02 and 4.91 mg/L for treatments A, B and C, respectively. Corresponding mean values of the area under the plasma concentration-time curve (AUC(0-infinity)) were 12.6, 12.6 and 14.2 mg/L.h, respectively. The mean half life (t1/2) values were 1.30, 1.23 and 1.50 hours for treatments A, B and C, respectively. Mean AUC(0-infinity), Cmax and t1/2 values following treatment B were not significantly different from those of treatment A. However, the mean AUC(0-infinity) and t1/2 values of cefprozil following treatment C were significantly greater than those of either treatment A or B. The mean Cmax value following treatment C was not significantly different than that of either treatments A or B. From these results, it was concluded that posture has no effect on the pharmacokinetics of cefprozil.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222463 TI - Drug dosage in end-stage renal disease (ESRD) patients undergoing haemodialysis. A predictive study based on a microcomputer program. AB - Drug dosage in end-stage renal disease (ESRD) patients undergoing haemodialysis is a very complex problem because of numerous variables relating to the patient, the type of drug administered and the type of dialysis and dialyser. We carried out a multifactorial study of these parameters using a microcomputer program written in BASIC. Three sets of data were fed into the computer: those relating to the biophysical characteristics of the patient, the type of dialysis and dialyser to be used, and others relating to the chemical and pharmacokinetic characteristics of the drug. From these, a predictive intravenous dosage regimen (bolus and infusion) was compiled for each ESRD patient. To check the program we used 2 drugs (tobramycin and vancomycin) and several types of dialyser. The findings were that, with tobramycin, an ESRD patient should be given different postdialytic doses, depending on the type of dialyser used. The maintenance doses calculated by the program were similar to those usually administered to patients receiving clinical treatment with this drug. In the case of vancomycin, the program calculated the clearance value in vivo through the 'Hemoflow F60' dialyser with a polysulphone membrane. The computer program calculated the maintenance dosage of vancomycin that should be given after each dialysis cycle so that its concentration did not fall below its minimum therapeutic concentration. PMID- 8222464 TI - Pregnancy in women with diabetes mellitus. The beginning. AB - Before the discovery of insulin in 1921, pregnancies in women with diabetes mellitus were uncommon. Most patients succumbed to ketoacidosis within 1 to 2 years after diagnosis. Insulin therapy restored the fertility of these women, and maternal deaths were nearly eliminated; however, the perinatal mortality rate remained high. Elective preterm deliveries were planned to reduce the stillbirth rate, often resulting in neonatal deaths from respiratory distress syndrome. Furthermore, women with more severe disease were able to become pregnant with a risk of complications due to preeclampsia. PMID- 8222465 TI - Metabolic fuel mixtures and diabetic embryopathy. AB - The incidence of congenital anomalies remains the major cause of morbidity and mortality among the offspring of diabetic women. Animal and human studies indicate that these malformations occur early in pregnancy and are influenced by the abnormal maternal metabolic milieu. This article reviews the recent advances in understanding the pathogenesis of diabetic embryopathy. A possible pathway to explain the various diabetic-related fuel aberration is presented. PMID- 8222466 TI - Prevention and diagnosis of congenital anomalies in diabetic pregnancies. AB - The high rate of major congenital malformations among infants of diabetic women represents the largest and most refractory problem in the perinatal care of these women. The prenatal diagnosis of these malformations will always be easier than their prevention. They are clearly not, however, entirely "beyond our therapeutic control." PMID- 8222467 TI - Fetal surveillance in the pregnancy complicated by diabetes mellitus. AB - During the last 15 years, protocols for antepartum fetal assessment in pregnancies complicated by diabetes mellitus have shifted the focus of care during the third trimester to an outpatient setting, reducing health care costs and emotional stress on patients and their families. Maternal assessment of fetal activity is the primary screening test in most surveillance programs, which use the nonstress test, contraction stress test, or biophysical profile. The success of these protocols continues to be predicted on careful regulation of maternal glycemia through aggressive therapy with insulin and diet. PMID- 8222468 TI - Maternal and perinatal implications of diabetic nephropathy. AB - Nephropathy is a serious and challenging complication of diabetes with regard to maternal health, fetal well-being, and outcome of infants. This article describes the available information and the controversies regarding clinical course, pregnancy complications, and treatment. PMID- 8222469 TI - Diabetic retinopathy during pregnancy. AB - This article outlines the pathophysiology of diabetic retinopathy, its epidemiology, and its diagnosis. Recommendations for management of the pregnant diabetic patient through the team approach has resulted in a dramatic improvement in maternal and fetal complications during pregnancy. By including a retinal specialist in the team during each trimester, the effects of pregnancy on the progression of diabetic retinopathy should be nearly identical to that of the non pregnant patient. PMID- 8222470 TI - Metabolic changes in gestational diabetes. AB - Normal pregnancy produces profound metabolic alterations that occur, in large part, by the conclusion of the first trimester. Although necessary to support the metabolic needs of the conceptus, the metabolic and cardiovascular changes of pregnancy are well established weeks before the metabolic needs of the fetus and placenta are fully expressed. The endocrine and metabolic characteristics of gestational diabetes, therefore, can only be understood against the backdrop of the rapidly changing conditions of normal pregnancy. Nondiabetic pregnancy produces an "accelerated starvation" in the fasting state with an earlier and more profound hypoglycemia and an increased fasting insulin. In the fed state, normal pregnancy produces a higher postprandial glycemic response despite an amplified first phase insulin response and higher plasma insulin concentrations. Both the intravenous glucose tolerance test and the steady state euglycemic clamp demonstrate that, by the second trimester, pregnancy produces peripheral and, perhaps, hepatic insulin resistance. The decreased peripheral uptake of glucose and the increased basal production of glucose in the fasting state support an increased flux of glucose to the fetus. Weight-matched obese gestational diabetic gravidae demonstrate a greater fall in fasting glucose and greater increase in ketosis compared with normal control gravidae. Obese GDM patients appear to have increased fasting insulin concentration compared with lean GDM and nondiabetic controls, whereas nonobese GDM patients do not appear to have elevated fasting insulin concentrations. Women with gestational diabetes have a greater rise in SI postpartum than controls, though the SI is the same in normal and gestational diabetic pregnancy in the third trimester.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222471 TI - Methods of screening for and diagnosing of gestational diabetes. AB - Screening for gestational diabetes should be performed using a 50-g 1-hour glucose challenge. The threshold for further testing may be chosen based on the goal of the screening program, either to maximize sensitivity at the expense of more diagnostic testing by using a 130 mg/dL cutoff or to increase specificity at the sacrifice of some sensitivity by using a 140 mg/dL cutoff. The choice of universal screening or an age cutoff, such as 25 years, with testing reserved in younger women for those with risk factors, is dependent on the characteristics of a given patient population or practice. The diagnostic test for gestational diabetes is the 100-g 3-hour oral glucose tolerance test on venous plasma or serum specimens. Whole blood values may be used, but the use of capillary blood measurements by test strips and reflectance meters is not recommended. PMID- 8222472 TI - Management of gestational diabetes. AB - Researchers and care providers must recognize that different centers are not automatically comparable in results because the geographic location, the demographic characteristics of the patient population, and the program strategy will alter the rate of patients assigned to insulin therapy and adverse pregnancy outcome. For example, in San Antonio, Texas, with a chronic health problem of obesity and diabetes, the prevalence of GDM is approximately 11%, whereas the general reported rate is 2% to 3%. Despite the disparity in the prevalence of the disease in different demographic areas, pregnancy outcome should be comparable for all centers when a proper management approach has been used. Utilizing our management approach in a large-scale program resulted in an incidence of macrosomia comparable to that in the general population. As a general rule, because constant evaluation of glycemia in pregnancy is the best gauge of the efficacy of the treatment, the use of verified self-monitoring of blood glucose has become a principal component of management. Appropriate assignment of patients to treatment modality utilizing verified blood glucose determinations and targeting mean blood glucose levels throughout pregnancy to 95 mg/dL (5.3 mmol/L) that is similar to normal nondiabetic blood glucose levels will result in pregnancy outcome comparable to the general population. PMID- 8222473 TI - Alternative therapies for the management of gestational diabetes. AB - The above is a review of current concepts and theories in GDM. The initial approach to the gestational diabetic women is to discover the problem early in pregnancy, before any sequelae occur. Traditionally, insulin was instituted early in the course of the gestational diabetic pregnancy. With appropriate monitoring, exercise and diet also may provide the mainstay of therapy. PMID- 8222474 TI - Infant of the diabetic mother. AB - Despite marked declines in neonatal mortality, infants of diabetic mothers still experience significant morbidity. This article reviews current data regarding congenital malformations, disorders of fetal growth, pulmonary development, and diagnosis and treatment of hypoglycemia, hypocalcemia, and hypomagnesemia. It also addresses short-term and long-term complications for this unique group of high-risk infants. PMID- 8222475 TI - Contraception in the diabetic woman. AB - Contraceptive prescription in women with current diabetes, type I or II, and in women with previous gestational diabetes must be highly effective and take into account the metabolic effects and risks particular to diabetes. This article concentrates on reversible, nonbarrier methods including oral contraceptives, long-acting progestins, and intrauterine devices with the goal of enabling the practitioner to develop individually tailored contraceptive therapy for these women. PMID- 8222476 TI - Alterations in theophylline metabolism during the first year of life. AB - Maturational changes in theophylline disposition were evaluated in 52 infants (gestational age, 24 to 40 weeks; postnatal age, 2 to 69 weeks) receiving maintenance theophylline therapy. Theophylline and metabolites were measured in serum and urine at steady state, and the influence of clinical parameters on the maturational changes was analyzed by multiple stepwise linear regression. Theophylline clearance and urine metabolite pattern reached adult values at 55 weeks' postconceptional age. Serum caffeine concentrations greater than 1 microgram/ml occurred in infants up to 50 weeks' postconceptional age. Disappearance of serum caffeine concentrations and maturation of theophylline clearance were primarily related (p < 0.001) to development of the demethylation pathway to 3-methylxanthine. Postconceptional age was the major factor (p < 0.001) explaining the interpatient variability in theophylline clearance (r2 = 0.57), serum caffeine to theophylline ratio (r2 = 0.46), and urinary excretion of theophylline (r2 = 0.51), caffeine (r2 = 0.49), 1,3-methyluric acid (r2 = 0.32), 1-methyluric acid (r2 = 0.53), and 3-methylxanthine (r2 = 0.58). Our findings indicate that postconceptional age rather than postnatal age should be used as a maturational marker during theophylline therapy in infancy. PMID- 8222477 TI - Colchicine disposition in human leukocytes after single and multiple oral administration. AB - Inasmuch as leukocytes were reported to be an active pharmacologic compartment, colchicine disposition was determined in plasma, granulocytes, and mononuclear cells in healthy volunteers after 1 mg oral single and multiple doses. After the single dose, maximal colchicine concentration was observed at 1 hour in plasma and 47 hours later in leukocytes. This delay was confirmed by the slow accumulation of colchicine by lymphocytes in culture. In the multiple-dose study, mean granulocyte colchicine concentration (20 to 53 ng/10(9) cells) were twofold higher than in mononuclear cells (9 to 24 ng/10(9) cells). Mean predicted colchicine multiple-dose granulocyte and mononuclear cell concentrations were 2.5 fold and ninefold higher, respectively, than those measured. After the last dose, colchicine decreased, with half-life values between 41 and 46 hours for leukocytes and 49 hours for plasma. This study validates leukocytes as a microcompartment whose kinetics correlates with colchicine biologic effects. PMID- 8222478 TI - Intravenous ciprofloxacin disposition in obesity. AB - The pharmacokinetics of ciprofloxacin and its metabolite 1 (desethyleneciprofloxacin) were studied in 17 obese men (mean age, 29.2 +/- 7.5 years; mean weight, 110.7 +/- 20.2 kg; mean body mass index, 36.4 +/- 3.9 kg/m2) and 11 control subjects (men of normal weight; mean age, 25.0 +/- 5.1 years; mean weight, 71.8 +/- 9.9 kg; mean body mass index, 23.3 +/- 2.4 kg/m2). Each subject received a single 400 mg intravenous dose of ciprofloxacin infused over 1 hour. Ciprofloxacin total clearance was significantly increased in obese subjects compared with control subjects (897.44 +/- 159.57 versus 744.44 +/- 120.51 ml/min, respectively; p < 0.05). Ciprofloxacin renal clearance in obese subjects (637.58 +/- 128.89 ml/min) was 29% higher than in control subjects (495.47 +/- 137.85 ml/min; p < 0.05). The elimination half-life values of ciprofloxacin and desethyleneciprofloxacin were not statistically different between groups. Ciprofloxacin steady-state volume of distribution (Vss) was significantly larger in obese group (269.17 +/- 51.64 versus 219.03 +/- 35.80 L; p < 0.01) compared with the control group, and when it was normalized by total body weight, obese subjects exhibited lower Vss/kg than control subjects (2.46 +/- 0.42 versus 3.06 +/- 0.31 L/kg; p < 0.001). These findings indicate that ciprofloxacin is distributed less to adipose tissue than to other tissues, but partial distribution to adipose tissue does occur. To normalize the volume of distribution of obese subjects to that of normal weight subjects, 45% of excess weight (total body weight minus ideal body weight) must be added to the ideal body weights of obese subjects. PMID- 8222479 TI - Pharmacokinetics and acute renal effects of continuously infused carboplatin. AB - Carboplatin was given as a 24-hour infusion at high doses to pediatric patients with cancer (n = 11) and pharmacokinetic parameters and renal effects were determined. Median carboplatin clearance for course 1 (151 ml/min per 1.73 m2;) was not significantly different from clearance for course 2 (144 ml/min per 1.73 m2; p = 0.33). The median glomerular filtration rate measured before (159 ml/min per 1.73 m2) and after course 1 (161 ml/min per 1.73 m2) did not differ significantly (p = 0.4). Binding was time dependent but modest with a median free fraction at the end of infusion of 0.82. Pharmacokinetic parameters for continuous-infusion carboplatin are similar to those reported for short infusions, but the median dose of 817 mg/m2 required to achieve an acceptable systemic exposure in these patients was 45% greater than the previously suggested maximum tolerated dosage. Continuous infusion carboplatin did not alter carboplatin clearance or adversely effect glomerular filtration rate during a second course, showing the feasibility of this alternative dosage strategy to enhance therapeutic effects. PMID- 8222480 TI - Method for studying drug-warfarin interactions. AB - The potential effects of extended-release felodipine on the pharmacokinetics and pharmacodynamics of warfarin were studied in a double-blind crossover study in 12 healthy men. Warfarin dosage was adjusted to achieve stable subtherapeutic anticoagulation. Subjects were then randomized to receive 2 weeks of treatment with 10 mg extended-release felodipine or placebo once daily, and warfarin dosage was adjusted if necessary to maintain stable international normalized ratio. The pharmacokinetics of R- and S-warfarin and the warfarin dose requirement did not differ importantly between periods of treatment with felodipine and placebo. The design of this study is suitable for general use in the identification of possible effects of other drugs on the pharmacokinetics and pharmacodynamics of warfarin. A different approach is needed if there is any reason to expect that warfarin may alter the pharmacokinetics or pharmacodynamics of the test drug. PMID- 8222481 TI - Disposition of digoxin immune Fab in patients with kidney failure. AB - Digoxin and digoxin immune Fab, its antidote, are eliminated renally. However, the disposition of Fab in severe kidney disease is poorly described. Therefore, the disposition of Fab and its relationship to total and free digoxin were studied in five digoxin-toxic patients with end-stage renal disease (n = 4) or severe renal dysfunction (n = 1) with a mean (+/- SD) serum creatinine of 5.9 +/- 1.2 mg/dl (four patients were receiving long-term hemodialysis). Serum was drawn after a clinically neutralizing Fab dose (80 to 160 mg) every 12 to 24 hours for 204 to 327 hours. Fab concentrations were assessed by radioimmunoassay, whereas total digoxin concentrations were assessed with a modified radioimmunoassay or fluorescence polarization immunoassay. The concentration-time profile of Fab appeared to be similar to the concentration-time profile of total digoxin. The mean (+/- SD) half-lives of the alpha and beta disposition phases of Fab were 13 +/- 5 hours and 96 +/- 31 hours, respectively, which were similar to the alpha and beta parameter estimates of total digoxin (14 +/- 4 and 123 +/- 16 hours, respectively). Steady-state volume of distribution and systemic clearance of Fab were 0.29 +/- 0.11 L/kg and 0.057 +/- 0.022 ml/min/kg, respectively. Thus, in comparison to values reported in patients with normal renal function, the elimination of Fab and total digoxin are markedly delayed in patients with end stage renal disease, which may necessitate prolonged clinical monitoring. PMID- 8222482 TI - Single-dose pharmacokinetics of cefpirome in patients receiving hemodialysis and in patients treated by continuous ambulatory peritoneal dialysis. AB - Cefpirome is eliminated primarily by renal excretion, compelling dosage reduction in kidney failure. We studied the elimination kinetics after intravenous administration of 1 gm cefpirome in patients undergoing hemodialysis (n = 9) and after intravenous (n = 6) or intraperitoneal administration (n = 6) of 1 gm cefpirome in subjects treated by continuous ambulatory peritoneal dialysis (CAPD). Four hours of hemodialysis removed 48% +/- 9% of the drug present in the body at the start of hemodialysis. Consequently, a supplementary dose equal to 50% of the daily dose recommended in end stage renal disease (ESRD) should be administered after each hemodialysis treatment. In patients receiving CAPD, therapeutic levels in both serum and dialysate were reached after intravenous and intraperitoneal administration. The bioavailability after intraperitoneal administration was 84%. After systemic administration, the elimination of cefpirome in the dialysate was negligible. Consequently, systemic dosage of cefpirome in patients receiving CAPD and in patients with ESRD should be identical. PMID- 8222484 TI - Tinnitus suppression by intravenous lidocaine in relation to its plasma concentration. AB - In a double-blind controlled trial in nine patients with tinnitus we measured the lidocaine plasma concentrations during and after intravenous administration of lidocaine or placebo and scored the level of tinnitus on a visual analog scale. No patient showed any effect during the placebo infusion. Administration of lidocaine resulted in total suppression or suppression to a non-annoying level of tinnitus in five patients, slight suppression but still annoying tinnitus in two patients, and worsening tinnitus in one patient. No effect of lidocaine was observed in one patient. Most relief was obtained at plasma concentrations between 1.5 and 2.5 micrograms/ml. In this concentration range a significant (p < 0.05) effect of lidocaine on tinnitus was observed. However, notable side effects were observed at plasma concentrations greater than 2.0 micrograms/ml. The effect persisted until plasma levels of about 0.5 microgram/ml were reached. A large variability in the effects existed because of variations in lidocaine kinetics and because of the presumed psychologic components of tinnitus. PMID- 8222483 TI - Gender-based effects on methylprednisolone pharmacokinetics and pharmacodynamics. AB - The pharmacokinetics and selected pharmacodynamic responses to methylprednisolone were investigated in six men and six premenopausal women after a dose of 0.6 mg/kg ideal body weight. Women (luteal phase) exhibited a greater methylprednisolone clearance (0.45 versus 0.29 L/hr/kg) and shorter elimination half-life (1.7 versus 2.6 hours) than men. The volume of distribution of methylprednisolone was similar when normalized for ideal body weight. Pharmacodynamic models were used to examine the methylprednisolone suppressive effects on cortisol secretion and basophil and helper T lymphocyte trafficking. A significantly smaller 50% inhibitory concentration (IC50) value (0.1 versus 1.7 ng/ml) was seen in the women for suppression of cortisol secretion, indicating increased sensitivity. However, the area under the concentration-time curve of effect was similar for both groups. The IC50 values for effects of methylprednisolone on basophil trafficking related to estradiol concentrations in a log-linear fashion in women, with increased sensitivity found at higher estradiol concentrations. Men displayed a greater 24-hour net suppression in blood basophil numbers, but no difference was observed in net cortisol and helper T lymphocyte suppression between the sexes. These findings suggest that methylprednisolone dosages should be based on ideal body weight. Although women are more sensitive to methylprednisolone as measured by cortisol suppression, they eliminate the drug more quickly, generally producing a similar net response. PMID- 8222485 TI - Modulation of vinblastine resistance with cyclosporine: a phase I study. AB - OBJECTIVE: Tumor cell resistance is a major cause of failure to cure advanced malignancies. Multidrug resistance is thought to be an important mechanism of such resistance. Our aims were to identify doses of cyclosporine that would achieve blood levels effective in modulating multidrug resistance to vinblastine and to evaluate the toxicities and maximum tolerated dose of cyclosporine when administered in conjunction with vinblastine. METHODS: We conducted a phase I trial of vinblastine and escalating doses of cyclosporine. Cyclosporine was given by continuous intravenous infusion over 120 hours and vinblastine was administered by continuous infusion from hour 12 to hour 108. Sixty-two patients entered the trial, of whom 60 were evaluable. RESULTS: Cyclosporine was escalated from 1 to 15.6 mg/kg/day. Vinblastine doses were reduced to 1.6 and then 1.2 mg/m2/day because of increasing vinblastine toxicity at higher cyclosporine doses. The maximum tolerated dose of cyclosporine at 1.2 mg/m2/day vinblastine was 12.5 mg/kg/day; at this dose level, mean blood cyclosporine level was 1.25 +/ 0.41 mumol/L. Significant nephrotoxicity was observed at higher cyclosporine doses in two of four patients. Nephrotoxicity was not significant at doses at or lower than this maximum tolerated dose and was not cyclosporine dose dependent. Myelosuppression, neurotoxicity, and transient hyperbilirubinemia were observed and were cyclosporine dose dependent. CONCLUSIONS. Cyclosporine by continuous infusion may be safely given in high doses concurrently with continuous-infusion vinblastine. Plasma levels of cyclosporine > or = 1 mumol/L can be sustained during vinblastine administration. No sustained effect on T-cell subsets was observed. Vinblastine toxicity is enhanced by cyclosporine in a dose-dependent fashion and correlates with cyclosporine-induced hyperbilirubinemia. PMID- 8222486 TI - Inhibition of leukotriene D4-induced bronchoconstriction in subjects with asthma: a concentration-effect study of ICI 204,219. AB - The peptide-leukotriene antagonist, ICI 204,219 [4-(5-cyclopentyloxycarbonylamino lmethylindol-3-ylmethyl )-3-methoxy-n-o-tolylsulfonyl benzamide], was administered 12 hours before an inhaled leukotriene D4 (LTD4) challenge during a double-blind, placebo-controlled, randomized, two-period crossover trial. Subjects with mild asthma were randomized into five treatment groups (six subjects each) and received single oral doses of placebo and either 5, 10, 20, 40, or 100 mg of ICI 204,219 on day 1 of each treatment period. ICI 204,219 was tolerated well by all subjects. A progressive dose response was observed for doses of ICI 204,219 from 5 mg through 100 mg. Compared with placebo, ICI 204,219 increased the concentration (PC20FEV1) and dose of LTD4 needed to reduce forced expiratory volume in 1 second (FEV1) by 20%. Mean LTD4 PC20FEV1 for groups that received placebo and 10, 40, or 100 mg ICI 204,219 increased by tenfold or more (p < 0.05). An association was found between the plasma concentration and protective effect of ICI 204,219 (p < 0.01). ICI 204,219 is the first leukotriene receptor antagonist for which a relationship has been established between drug plasma levels and its protective effect in subjects with asthma. PMID- 8222487 TI - High-density lipoprotein cholesterol elevation with gemfibrozil: effects of baseline level and modifying factors. AB - The effects of baseline level and modifying factors on gemfibrozil-induced high density lipoprotein (HDL) cholesterol elevation were studied in 1028 participants with good compliance in the Helsinki Heart Study. The absolute (mmol/L) increment in HDL cholesterol was independent of baseline when the change in the placebo group (regression toward the mean) was considered. In contrast, absolute reductions in low-density lipoprotein (LDL) cholesterol and triglycerides correlated with their baselines, relative percentage changes being constant. Statistically, this could indicate differences in the mode of action of gemfibrozil: an independent and additive effect on HDL cholesterol and a multiplicative effect on LDL cholesterol and triglycerides. These differences may have a physiologic background because the main effect of gemfibrozil is in the stable HDL3 subfraction, rather than in the variable HDL2. Only 13% of the variation in gemfibrozil-induced HDL cholesterol changes were explained by modifying factors. The basic assumptions in the uses of absolute or relative changes as a measure of treatment effect are discussed. PMID- 8222488 TI - Prednicarbate activity and benefit/risk ratio in relation to other topical glucocorticoids. AB - BACKGROUND: Nonhalogenated double esters of prednisolone or hydrocortisone applied topically to the skin have a low atrophogenic potential. However, activity and benefit/risk ratio and therefore the superiority over conventional topical glucocorticoids are not well defined. METHODS: The activities of cream preparations with prednicarbate (0.025% to 0.25%), hydrocortisone aceponate, and hydrocortisone buteprate (0.1%) are compared to the effects of betamethasone 17 valerate (0.1%), hydrocortisone (1%), and two drug-free vehicles in 60 healthy volunteers. Test models are the skin blanching assay (occluded and nonoccluded mode), ultraviolet-induced erythema, and an irritant (sodium dodecyl sulfate) dermatitis. The benefit/risk ratio is derived from the activity in the former models and the reduction of skin thickness as determined previously. RESULTS: Prednicarbate activity increases in a dose-dependent manner. Prednicarbate, 0.25%, and the hydrocortisone double esters appear to be equipotent to betamethasone 17-valerate in the skin blanching test and the ultraviolet-erythema test, but superior to hydrocortisone and the vehicles. Prednicarbate and its vehicle, however, do not reverse irritant dermatitis. The benefit/risk ratios of prednicarbate and hydrocortisone aceponate exceed those with betamethasone 17 valerate. CONCLUSIONS: Prednicarbate and hydrocortisone aceponate are intermediate potent glucocorticoids that are superior to betamethasone 17 valerate because of the improved benefit/risk ratio. Patients with severe atopic dermatitis and those who relapse frequently should profit from the treatment with these newer glucocorticoids. PMID- 8222489 TI - Impairment of cefdinir absorption by iron ion. AB - The effect of iron ion on the absorption of cefdinir, a new oral cephalosporin derivative, was evaluated in healthy male volunteers in a randomized three-way crossover study. The subjects received 200 mg cefdinir alone, 200 mg cefdinir and two tablets of iron ion concomitantly, and two tablets of iron ion preparation 3 hours after 200 mg cefdinir administration. The area under the concentration curve [AUC(0-12)] of cefdinir with concurrent iron was significantly smaller than that with cefdinir alone (mean +/- SD, 0.78 +/- 0.38 versus 10.3 +/- 1.35 micrograms.hr/ml). While there were no differences in AUC(0-3) between drug alone and drug with iron 3 hours later, the AUC(3-12) with delayed iron was significantly smaller than that of cefdinir alone (4.60 +/- 1.54 versus 8.03 +/- 1.72 micrograms.hr/ml). These findings suggest that the mechanism of interaction between cefdinir and iron ion preparation is the formation of a chelation complex and that this complex probably restricts gastrointestinal absorption. PMID- 8222490 TI - Role of intrahepatic protein binding in the hepatobiliary extraction profile of cefixime in humans. AB - The hepatobiliary extraction profile of cefixime, a dianionic cephalosporin antibiotic, was studied in 10 patients, each of whom was provided with T-tube drainage of his or her common bile duct after cholecystectomy. After a single 200 mg oral dose, cefixime biliary clearance proved to be nonlinear, mostly in its initial phase, which is consistent with a concentrative uptake and intracellular protein binding for the drug. The latter process appears to be saturable and to operate at a rate that correlates with the total amount of cefixime recovered in the 24-hour bile drainage. Such findings seem to confirm the significant role played in vivo by hepatic ligandin in the hepatobiliary extraction of organic anions. The data also show that a single 200 mg oral dose of cefixime yields drug levels in bile substantially higher than the minimal inhibitory concentrations for the most frequent Enterobacteriaceae in biliary tract infections. Accordingly, this cephalosporin could be an interesting alternative in both prophylaxis and treatment of biliary tract infections. PMID- 8222491 TI - In vivo and in vitro correlation of microsomal epoxide hydrolase inhibition by progabide. AB - Progabide was investigated as a potential inhibitor of microsomal epoxide hydrolase as a result of reports of elevated levels of carbamazepine-10,11 epoxide after coadministration of progabide and carbamazepine to patients with epilepsy. The formation clearance of carbamazepine transdihydrodiol after administration of carbamazepine-10,11-epoxide to healthy volunteers was decreased 26% by progabide. Therapeutic concentrations of progabide inhibited S (+)-styrene oxide hydrolysis in human liver microsomes (inhibition constant [Ki] = 1.9 mumol/L) and purified human liver microsomal epoxide hydrolase (Ki = 4.4 mumol/L). A mixed competitive and noncompetitive mechanism of inhibition best described the effect of progabide on microsomal epoxide hydrolase; the most potent inhibition was competitive. A similar model described the inhibition by the acid metabolite of progabide, although inhibitory concentrations are higher than concentrations observed after progabide therapy. An excellent agreement between the in vivo and in vitro inhibitory potencies of progabide suggests that potential inhibitors of this important detoxification enzyme can be predicted in vitro. PMID- 8222492 TI - Human liver dehydroepiandrosterone sulfotransferase: nature and extent of individual variation. AB - Dehydroepiandrosterone sulfotransferase (DHEA ST) catalyzes the sulfation of steroid hormones such as DHEA, estrone, and estradiol. As a first step in pharmacogenetic studies of DHEA ST in humans, we measured individual variation in DHEA ST enzymatic activity and thermal stability in 94 samples of human hepatic tissue, 39 of which were from patients with normal liver function studies. Neither level of enzyme activity nor thermal stability were significantly correlated with either time of tissue storage at -80 degrees C or patient age. In addition, there were no gender-dependent differences in DHEA ST activity in these samples. DHEA ST enzymatic activity varied 4.6-fold, with a mean value of 317 +/- 100 units/gm tissue (mean +/- SD) in all samples and 318 +/- 104 units/gm in the subset of 39 samples from patients with normal hepatic function studies. Frequency distributions of DHEA ST activity for both the entire group of 94 samples and the subset of 39 were bimodal, with 25% and 21% included in a high activity subgroup, respectively. The presence of this high activity subgroup was confirmed when data for samples from male and female patients were evaluated separately and when only data for white patients were examined. The existence of a subgroup of subjects with a high level of DHEA ST enzymatic activity in liver and a 4.6-fold range in this activity have implications for individual differences in the sulfate conjugation of endogenous and exogenously administered steroid hormones and raise the possibility of pharmacogenetic regulation of this important enzyme in humans. PMID- 8222493 TI - Ethnic differences in response to morphine. AB - Only recently has attention been focused on the importance of interethnic differences as determinants of interindividual variability in drug response. We compared the pharmacokinetics and pharmacodynamics of morphine in eight Chinese and eight white healthy mean after 0.15 mg/kg of morphine intravenously. The clearance of morphine was significantly higher in the Chinese subjects than in the white subjects because of an increase in the partial metabolic clearance by glucuronidation. There was no interethnic difference in the metabolism to normorphine. Morphine depressed the respiratory response to rebreathing carbon dioxide more in white subjects than in Chinese subjects, resulting in a greater reduction in resting ventilation and resting end-tidal PCO2. The slope of the ventilation/PCO2 response curve, a measure of carbon dioxide sensitivity, was reduced more in white subjects than Chinese subjects. As a result, white subjects had a greater depression in ventilation at a PCO2 of 55 mm Hg. The morphine induced reduction in blood pressure was also greater in white subjects than in Chinese subjects. Thus this study has shown ethnicity to be an important determinant of the disposition and effects of morphine. PMID- 8222494 TI - The differential effects of circulating norepinephrine and neuronally released norepinephrine on sodium excretion in humans. AB - The renal effects of incremental infusions of norepinephrine (placebo, 0.025 mu/kg/min), 0.075 micrograms/kg/min, phenylephrine (placebo, 0.5 micrograms/kg/min, 2.5 micrograms/kg/min), and tyramine (placebo, 2 micrograms/kg/min, 15 micrograms/kg/min) were examined in three respective groups (n = 9, 8, and 8) of normotensive male subjects undergoing water diuresis. Tyramine is an indirect sympathetic agent that causes neuronal release of endogenous norepinephrine. Increases in mean arterial pressure during each high dose infusion were comparable in all three groups. Both norepinephrine and phenylephrine caused a decrease in urinary sodium excretion and effective renal plasma flow, with no changes in glomerular filtration rate. Proximal tubular sodium reabsorption, as assessed by both lithium clearance and solute-free water clearance methods, was increased by pressor doses of norepinephrine and phenylephrine. In contrast, a similar pressor dose of tyramine was associated with a pressure natriuresis, an increase in effective renal plasma flow, and a decrease in proximal tubular sodium reabsorption. Our data indicate that, in normotensive humans, circulating catecholamines (norepinephrine and phenylephrine) have opposite effects on renal sodium handling from neuronally released norepinephrine (tyramine). PMID- 8222495 TI - The hypotensive action of captopril and enalapril is not prostacyclin dependent. AB - Angiotensin converting enzyme inhibitors have been proposed to have a prostaglandin-dependent component to their hypotensive action. The aim of this study was to assess whether the structurally dissimilar angiotensin converting enzyme inhibitors captopril and enalapril stimulate the synthesis of prostacyclin, whether their hypotensive action is blunted by indomethacin, and whether these biochemical or physiologic parameters differ for the two drugs, in white subjects with essential hypertension. Twelve patients were enrolled and 11 finished the study. The study consisted of a double blind, randomized, double crossover design. All patients received either placebo or 50 mg indomethacin twice a day for 3 weeks; after 1 week of placebo or indomethacin either 50 mg captopril or 10 mg enalapril twice a day was added and continued for 2 weeks. Each patient received every possible combination. Neither captopril nor enalapril stimulated prostacyclin production as determined by measurement of the urinary excretion rate of its main enzymatic metabolite, 2,3-dinor-6-keto-prostaglandin F1 alpha. Although indomethacin reduced the urinary excretion of the enzymatic metabolite of prostacyclin by more than 50%, it did not influence the hypotensive effect of captopril or enalapril. We conclude that neither captopril nor enalapril have a significant prostacyclin-dependent component to their hypotensive action. PMID- 8222496 TI - The gastrin-receptor antagonist L-365,260 inhibits stimulated acid secretion in humans. AB - We investigated the effect of a novel gastrin-cholecystokinin-B receptor antagonist, L-365,260 [(3R)-3(N'-3-methylphenyl)ureido)-1,3-dihydro-5-phenyl- 2H 1,4-benzodiazepin-2-one], on gastric acid secretion in humans. In a double-blind, four-period crossover study, eight subjects received single oral doses of placebo or of 2.5, 10, or 50 mg L-365,260, followed by an intravenous infusion of pentagastrin at doses of 0.05, 0.4, and 2 micrograms/kg/hr for successive 30 minute periods. L-365,260 caused a dose-dependent inhibition of pentagastrin stimulated gastric acid secretion. A single oral dose of 50 mg L-365,260 produced 50% inhibition of the gastric acid output response to pentagastrin (0.4 micrograms/kg/hr) when the mean (+/- SD) plasma L-365,260 concentration was 502 +/- 108 ng/ml. Plasma L-365,260 concentrations (all doses combined) and the inhibition of gastric acid output were correlated with a correlation coefficient of r = 0.45 (p < 0.05). Single oral doses of L-365,260 up to 50 mg did not inhibit basal gastric acid output or alter plasma gastrin concentrations. L 365,260 was well tolerated at oral doses up to 50 mg. These findings show that L 365,260 is an orally active antagonist at gastrin-cholecystokinin-B receptors in humans. PMID- 8222497 TI - Comparative study on the efficacy, acceptability, and side effects of a contraceptive pill administered by the oral and the vaginal route: an international multicenter clinical trial. AB - The objective of this multicenter randomized clinical trial was to compare the efficacy, acceptability, and occurrence of side effects associated with the oral versus vaginal route of administration of contraceptive pills. Eight hundred nineteen healthy, parous women of reproductive age were recruited at family planning clinics and research centers, members of the South to South Cooperation in Reproductive Health, in seven countries of the developing world. These women were randomly assigned to use either oral or vaginal administration of the same contraceptive pill, which contained 250 micrograms levonorgestrel and 50 micrograms ethinyl estradiol. No statistically significant differences were found in discontinuation rates between the two groups after 1 year. Involuntary pregnancy rates after 1 year were not statistically significantly different between the two groups. The vaginal route of administration appears to be as acceptable and efficacious as the oral route. PMID- 8222498 TI - Ketoprofen, acetaminophen plus oxycodone, and acetaminophen in the relief of postoperative pain. AB - Ketoprofen (Orudis) is a nonsteroidal anti-inflammatory drug that is currently approved in the United States for the management of mild to moderate pain. The objective of this trial was to determine the effectiveness of orally administered ketoprofen in the management of severe postoperative pain. This randomized, double-blind parallel study compared the efficacy and safety of single doses of 100 mg or 50 mg ketoprofen, the combination of 650 mg acetaminophen plus 10 mg oxycodone hydrochloride, 650 mg acetaminophen, or placebo in 240 patients with severe postoperative pain after cesarean section. Analgesia for the first dose was assessed over an 8-hour period. Multiple doses of 100 mg or 50 mg ketoprofen and the combination at half the dose (325 mg acetaminophen plus 5 mg oxycodone) were also assessed for up to 7 days. The 100 and 50 mg doses of ketoprofen and the combination were statistically superior to acetaminophen and placebo for many analgesic measures. A dose response was observed between the two doses of ketoprofen, with the 100 mg dose providing significantly greater analgesia over the lower dose. Ketoprofen, 100 mg, was at least as effective as the combination and its effects lasted longer, with the exception of hour 1 when the combination was superior. Remedication time for the group receiving 100 mg ketoprofen was significantly longer than for the other treatment groups. Significantly more patients who took repeated doses of the combination (84%) than those who took either dose of ketoprofen (70%) had adverse effects. Ketoprofen at both dose levels was shown to be effective, long-lasting, and well tolerated, and it should be considered as a viable option for the management of moderate to severe postoperative pain. PMID- 8222500 TI - The placebo effect in healthy volunteers: influence of experimental conditions on the adverse events profile during phase I studies. AB - In contrast to the plethora of publications on placebo effects in patients, very little is known about placebo effects in healthy volunteers during clinical pharmacology studies. We therefore reviewed the adverse events spontaneously reported during placebo administration in 109 double-blind, placebo-controlled studies involving 1228 volunteers. The overall incidence of adverse events in the healthy volunteers during placebo administration was 19%. As expected, complaints were more frequent after repeated dosing (28%) and in elderly subjects (26%). Overall, the most frequent adverse events were headache (7%), drowsiness (5%), and asthenia (4%), with some variation depending on study design and population. In conclusion, these data shed new light on the impact of experimental conditions on the results of safety evaluations in healthy volunteers participating in clinical pharmacology studies. PMID- 8222499 TI - Efficacy and tolerability of the renin inhibitor Ro 42-5892 in patients with hypertension. AB - The efficacy of multiple oral administration of the renin inhibitor Ro 42-5892 [(S)-alpha-](t-butylsulfonyl)-methyl]hydrocinnamamido]-N-[1S , 2R,3S)-1 (cyclohexylmethyl)-3-cyclopropyl-2,3-dihydroxypropyl]-imi dazole-4- propionamide] was studied. Forty-nine patients with moderate essential hypertension were randomly assigned to three groups that entered an 8-day double-blind oral treatment period: daily administration of placebo (group A), 300 mg Ro 42-5892 (group B), or 600 mg Ro 42-5892 (group C). Four hours after the last oral drug intake, placebo was administered intravenously to subjects in group A and 100 mg Ro 42-5892 was administered intravenously to subjects in groups B and C. Sitting systolic and diastolic blood pressures were measured on days 1 and 8 with a blood pressure device. On day 1, systolic blood pressure maximally decreased by 13.3 +/ 9.3, 20.2 +/- 11.2, and 24.1 +/- 11.3 mm Hg in groups A, B, and C, respectively (mean +/- SD; p < 0.01 for group A versus group C). Diastolic blood pressure maximally decreased 9.4 +/- 5.7, 13.9 +/- 8.7, and 11.8 +/- 5.7 mm Hg (difference not significant). On day 8, systolic blood pressure maximally decreased 19.5 +/- 16.5, 26.5 +/- 17.4, and 30.5 +/- 18.4 mm Hg and diastolic blood pressure maximally decreased 14.8 +/- 5.0, 16.2 +/- 9.0, and 17.9 +/- 12.7 mm Hg (difference not significant) compared with pretreatment values. Intravenous drug administration did not further reduce blood pressure, suggesting that the mode of action and not the low bioavailability was the limiting factor for the low efficacy. PMID- 8222501 TI - Interpretation of oxygen and carbon dioxide exchange across tissue beds in vivo. PMID- 8222502 TI - A new calcitonin-receptor-like sequence in rat pulmonary blood vessels. AB - 1. Two rat clones have been isolated which are similar to known calcitonin receptor sequences. One of these does not have the distribution expected of a calcitonin receptor. It is widely distributed, with extremely high levels of expression in the lung, where it is associated with the blood vessels. 2. This rat sequence may represent the receptor for calcitonin-gene-related peptide or islet amyloid polypeptide. Both have binding activity in the lung and are potent vasodilators. The gene represented by this sequence may therefore play an important role in the maintenance of vascular tone. PMID- 8222503 TI - Effect of inhalation of 30% nitrous oxide on spectral components of heart rate variability in conscious man. AB - 1. Periodicities in cardiac interbeat interval may be resolved into discrete frequency components by applying Fourier analysis to heart rate time series. Low frequency components (< 0.15 Hz) are believed to be under parasympathetic and sympathetic control, whereas a higher frequency component in phase with respiration is believed to be entirely parasympathetic. The ratio of the power in the low-/high-frequency spectrum gives an estimate of sympathetic/parasympathetic balance. 2. This study examined, using heart rate variability spectral analysis, the cardiac autonomic effects of breathing 30% N2O in normal subjects. While supine, the inhalation of N2O caused a significant fall in high-frequency power and a rise in the low-/high-frequency spectrum. During air breathing, tilting caused a significant rise in the mean blood pressure, heart rate, low-frequency power and low-/high-frequency spectrum. During N2O breathing, tilting caused a rise in the heart rate and the mean blood pressure, but no significant alteration in the power of individual spectral components. During tilting, the heart rate, the low-frequency and low-/high-frequency spectrum were less when breathing N2O than when breathing air. 3. These observations are consistent with the effect of N2O being an enhanced sympathetic balance of sinoatrial control, with the primary effect being through reduced parasympathetic tone. Enhanced sympathetic dominance of heart rate variability was seen on standing while subjects breathed air, but this effect was blunted with N2O. PMID- 8222504 TI - Cardiovascular and sympatho-adrenal responses to mental stress in primary hypertension. AB - 1. Eleven untreated men with mild to moderate primary hypertension and 10 normotensive control subjects were studied at rest and during a mental stress test (Stroop colour word conflict test), which has previously been used in studies of hypertensive patients with regard to non-invasive cardiovascular variables and venous plasma catecholamine concentrations. 2. Heart rate, central cardiovascular pressures, cardiac output (thermodilution) and forearm blood flow (strain gauge plethysmography) were determined. Systemic and forearm vascular resistances were calculated. Arterial and venous plasma adrenaline and noradrenaline concentrations were measured by h.p.l.c., and arterial noradrenaline spillover and noradrenaline overflow from the forearm were assessed by isotope methodology ([3H]noradrenaline). Neuropeptide Y-like immunoreactivity was measured by radioimmunoassay. 3. In hypertensive patients heart rate, arterial blood pressure, cardiac output and forearm blood flow increased by 28%, 13%, 37% and 115%, respectively, and forearm and systemic vascular resistances decreased by 48% and 21%, respectively (P < 0.001 for all responses), during stress. These responses were not different from those of the control group. 4. Arterial noradrenaline spillover rose by 63% and noradrenaline overflow from the forearm rose by 150% in the hypertensive patients in response to mental stress (P < 0.001); no significant group differences could be demonstrated. However, the forearm noradrenaline overflow response to stress tended to be greater in the hypertensive group (P = 0.11). Arterial adrenaline concentrations doubled in both groups (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222505 TI - Plasma levels of brain natriuretic peptide in healthy subjects and patients with essential hypertension: response to posture. AB - 1. To examine whether posture-induced changes in central volume affect brain natriuretic peptide secretion, plasma levels of human brain natriuretic peptide 32-like immunoreactivity (hBNP-32-li) were measured by radioimmunoassay in 11 healthy subjects and 20 patients with essential hypertension after 15 min supine, 15 min sitting and 15 min with the legs raised at 60 degrees, together with plasma atrial natriuretic peptide concentration, plasma renin activity and plasma aldosterone concentration. 2. In the supine position, the plasma hBNP-32-li level was 1.57 +/- 0.10 fmol/ml in healthy subjects and significantly higher in hypertensive patients (2.39 +/- 0.13 fmol/ml, P < 0.001). In both groups, plasma hBNP-32-li level significantly (P < 0.001) decreased when sitting (normotensive, 1.22 +/- 0.08 fmol/ml; hypertensive, 1.85 +/- 0.15 fmol/ml, P < 0.001 versus normotensive) and increased again after leg raising (normotensive, 2.13 +/- 0.12 fmol/ml; P < 0.002 versus resting; hypertensive, 2.84 +/- 0.16 fmol/min, P < 0.001 versus resting, P < 0.025 versus normotensive). 3. The plasma atrial natriuretic peptide concentration showed similar behaviour to the plasma hBNP-32 li, whereas plasma renin activity and plasma aldosterone concentration increased during sitting and decreased during leg raising in both healthy subjects and hypertensive patients, who had significantly higher plasma aldosterone levels when supine and sitting.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222506 TI - Expression of the three endothelin genes and plasma levels of endothelin in pre eclamptic and normal gestations. AB - 1. Maternal vasoconstriction and fetal growth retardation are part of the syndrome of pre-eclampsia and are thought to be related to placental insufficiency. There is evidence to suggest that the powerfully vasoactive endothelin peptides might be involved in the pathogenesis. 2. The production of endothelins appears to be regulated mainly by modulation of mRNA levels, and they are thought to exert their effects locally, rather than systemically. Hence, the measurement of endothelin mRNA levels in tissues would be expected to reflect the activity of the endothelin systems. 3. RNAase protection assays, using specific antisense probes capable of distinguishing between the endothelin mRNA isoforms, have been used to examine the expression of the three endothelin genes in placental villous tissue, amniotic membrane and myometrium. Samples were taken from 15 pre-eclamptic women and from 14 women with normal gestations. Myometrium was taken at hysterectomy from five non-pregnant women as an additional control. Plasma concentrations of immunoreactive endothelin [endothelin-1, endothelin-2 and big endothelin (not distinguished)] were measured in samples taken concurrently from uterine vein and peripheral blood during surgery in ten patients. 4. The level of endothelin-1 mRNA in placental villous tissue was significantly higher in pre-eclamptic women (32 weeks gestation) than in control subjects (38 weeks gestation) (1.85 +/- 0.26 versus 0.52 +/- 0.09 arbitrary units, means +/- SEM, P < 0.001) and in gravid than in non-gravid myometrium (P < 0.001). Endothelin-2 was not expressed in villous tissue, but was expressed in amniotic membrane and myometrium, with relatively little difference between pre eclamptic and control groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222507 TI - Concentration-dependent effects of insulin on Ca2+ influx in vascular smooth muscle cells of normotensive and spontaneously hypertensive rats. AB - 1. The effect of insulin on cytosolic free Ca2+ concentration was measured using fura-2 in vascular smooth muscle cells of normotensive and spontaneously hypertensive rats. 2. In both strains, insulin increased cytosolic free Ca2+ concentration in a concentration range between 10(-6) and 10(-3) units/ml. The maximum increase in cytosolic free Ca2+ concentration was observed with 10(-5) units/ml insulin (107 +/- 25 and 82 +/- 27 nmol/l in spontaneously hypertensive rats and normotensive rats, respectively). 3. The effect of insulin was dependent on extracellular Ca2+ and was enhanced by stimulation of protein kinase C. 4. Thus insulin appears to induce a Ca2+ influx in vascular smooth muscle cells only over a certain range of concentrations. No significant difference in the response to insulin of cells from normotensive and hypertensive rats was observed. PMID- 8222508 TI - Effects of angiotensin II on insulin sensitivity: a placebo-controlled study. AB - 1. There is evidence that hyperinsulinaemia increases the aldosterone response to angiotensin II, and that angiotensin-converting enzyme inhibitor drugs enhance peripheral glucose utilization, but the direct effects of angiotensin II on insulin sensitivity have not been reported previously. 2. In a randomized, double blind, placebo-controlled, cross-over study, 12 healthy male subjects attended on 3 study days for the evaluation of the effects of a subpressor (1 ng min-1kg-1) and pressor (5 ng min-1kg-1) infusion of angiotensin II on whole-body insulin sensitivity using the euglycaemic hyperinsulinaemic clamp. Frequent measurements of blood pressure and heart rate were recorded and blood samples were collected for determination of serum insulin, C-peptide and K+ concentration, plasma renin activity and plasma angiotensin II concentration. 3. Plasma angiotensin II concentrations (means +/- SD) were 11 +/- 5 pg/ml after placebo, and 27 +/- 9 and 125 +/- 28 pg/ml after low and high dose angiotensin II, respectively. The higher dose of angiotensin II was associated with significant increases in blood pressure (e.g. 13 mmHg systolic blood pressure at 150 min) and serum aldosterone concentration. Whole-body insulin sensitivity was 10.5 +/- 2 mg of glucose min 1kg-1 after placebo, and 10.5 +/- 2.2 and 10.9 +/- 3.4 mg of glucose min-1kg-1 after low and high dose angiotensin II (not significant). 4. Angiotensin II had no effect on hyperinsulinaemia-induced reductions in serum potassium and triacylglycerol concentrations. 5. Thus, acute infusion of angiotensin II for 3 h, with or without an increase in blood pressure, has no effect on whole-body insulin sensitivity. PMID- 8222509 TI - Effects of decreased glutamine supply on gut and liver metabolism in vivo in rats. AB - 1. It has recently been suggested that glutamine may be a conditionally essential nutrient rather than a non-essential amino acid. Therefore, administration of methionine sulphoximine was used to create a model of decreased arterial glutamine concentrations for 4 days. Glutamine consumption in portal-drained viscera and liver was measured after an overnight fast by determining fluxes and intracellular concentrations in normal rats, methionine sulphoximine-treated rats and pair-fed controls. Moreover, fluxes and intracellular concentrations of several other amino acids and ammonia and production of urea by the liver were determined concomitantly. 2. Methionine sulphoximine treatment for 4 days resulted in a 50% decrease in arterial glutamine concentration. Although the glutamine consumption and the intracellular glutamine concentration of the intestine were reduced by 50% at day 4, no changes in intestinal amino acid and ammonia metabolism were observed. 3. In the liver, glutamine consumption was reduced and ammonia uptake was increased, but urea synthesis was not changed. The decreased intracellular glutamine, glutamate, aspartate and ammonia concentrations coincided with a substantial reduction in liver branched-chain amino acid production. 4. These results suggest that reduced intestinal glutamine uptake does not induce marked changes in intestinal amino acid metabolism. The decreased liver branched-chain amino acid production suggests a reduction in the net liver protein degradation rate during methionine sulphoximine treatment. PMID- 8222510 TI - Effects of acid loading on serum amino acid profiles and muscle composition in normal fed rats. AB - 1. Impaired body growth, loss of lean body mass and negative nitrogen balance are common findings in chronic renal failure. Enhanced endogenous protein catabolism in skeletal muscle induced by co-existent metabolic acidosis seems to be an important aetiological factor. 2. In uraemic patients abnormalities in blood amino acid profiles are present after a protein meal, suggesting impaired metabolism of ingested proteins. 3. This study demonstrates that acidosis induces changes in arterial amino acid profiles in fed non-uraemic rats. These included increased levels of threonine, histidine, proline, serine and glycine, and decreased levels of tryptophan. These changes are similar to those found in uraemic patients after a protein meal, suggesting a pathogenic role of acidosis in the impairment of exogenous protein metabolism. 4. Intracellular amino acid levels in the muscle tissue partly reflect the changes in the extracellular level. 5. Acidotic animals had a decreased body weight gain and a reduced alkali soluble protein/DNA ratio in muscle cells compared with controls. 6. In conclusion, the results show that acidosis per se modifies the circulating amino acid profile in fed rats, resulting in a pattern similar to the post-prandial amino acid changes described in uraemic patients. These abnormalities occur together with impaired growth of skeletal muscle cells. PMID- 8222511 TI - Determination of free polyamines in human bile by high-performance liquid chromatography. AB - 1. Polyamines are widely distributed in the body and may have cholesterol nucleating activity in model bile and human bile. There are only partial and scant data available on the type of polyamines in human bile. 2. In this study methods for extraction of free polyamines, benzoylation and an h.p.l.c.-based method for the quantitative determination of putrescine, cadaverine, spermidine and spermine in bile are described. The h.p.l.c. methodology was validated and compared with separation on t.l.c. after dansylation. 3. The polyamine content of 11 gallbladder bile samples and 11 T-tube bile samples was analysed, all from patients with gallstones. Polyamines were found in three out of 11 gallbladder bile samples and eight out of 11 T-tube bile samples. Putrescine levels were 0.5 287 mumol/l and cadaverine levels were 2.4-645.4 mumol/l; these were considerably higher than spermine and spermidine levels. 4. As many of these gallstones bile samples were devoid of polyamines, it is questionable whether polyamines play an important role in cholesterol nucleation in human bile. PMID- 8222512 TI - Acetyl-CoA:lyso-platelet-activating factor acetyltransferase activity in neutrophils from asthmatic patients and normal subjects. AB - 1. Platelet-activating factor is a putative mediator of inflammation in asthma and the enzyme acetyl-CoA:lyso-platelet-activating factor acetyltransferase appears to be important in regulating platelet-activating factor production by leucocytes. To determine whether there are differences in acetyltransferase activity between asthmatic patients and normal subjects, enzyme activity was assayed in neutrophil lysates from atopic asthmatic patients (n = 20), aspirin sensitive asthmatic patients (n = 12) and healthy, non-atopic, non-asthmatic control subjects (n = 20), both basally and after stimulation with the calcium ionophore A23187. 2. For a range of acetyl-CoA concentrations, acetyltransferase activity (nmol of [acetyl-3H]PAF min-1 mg-1 of protein) in unstimulated neutrophils from atopic asthmatic patients was significantly higher than that for normal subjects (P = 0.038) and the mean Vmax. for atopic asthmatic patients [18.4 (SD 6.9) nmol min-1 mg-1 of protein] was significantly greater than that for the control subjects [14.9 (SD 4.6) nmol min-1 mg-1 of protein P < 0.05]. The mean Vmax. for aspirin-sensitive asthmatic patients [15.9 (SD 6.9) nmol min-1 mg 1 of protein] was not significantly different from that for the normal subjects. 3. The mean ratio Vmax. stimulated/Vmax. unstimulated for acetyltransferase from atopic asthmatic patients (1.71, SD 0.45) was significantly less than that for the normal subjects (2.13, SD 0.63, P < 0.05), suggesting that acetyltransferase from atopic asthmatic patients was less sensitive to stimulation with A23187 in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222513 TI - Effect of chronic low-dose arginine vasopressin infusion on body fluid homoeostasis during adaptation from a high- to a low-sodium diet in normal man. AB - 1. A diuresis occurs within the first 36h of salt restriction. A decline in plasma arginine vasopressin concentration may contribute to both the diuresis and antinatriuresis. 2. We have studied six normal human subjects during 36h of dietary sodium restriction. In one study subjects received an intravenous infusion of D-glucose, and in the other an infusion of arginine vasopressin (6 fmol min-1 kg-1). 3. In the D-glucose phase plasma arginine vasopressin concentration fell (1.77 +/- 0.34 to 1.02 +/- 0.13 pg/ml), urine flow increased (67.9 +/- 11.5 to 89.8 +/- 17.1 ml/h), haemoconcentration occurred (packed cell volume 40.8 +/- 0.3 to 42.8 +/- 0.5%, protein concentration 71.6 +/- 0.5 to 74.5 +/- 0.6 g/l), plasma sodium concentration fell (140 +/- 0.2 to 138 +/- 0.2 mmol/l) and plasma renin activity increased (1600 +/- 153 to 3700 +/- 356 pg of angiotensin I h-1 ml-1). 4. In the arginine vasopressin phase plasma arginine vasopressin concentration remained constant (1.5 +/- 0.13 to 1.34 +/- 0.11 pg/ml), the diuresis was reversed (65.7 +/- 9.9 to 52.1 +/- 8.9 ml/h), plasma sodium concentration fell further (139.8 +/- 0.4 to 136.1 +/- 0.4 mmol/l), the rise in plasma renin activity was reduced (arginine vasopressin 2552 +/- 292; D glucose, 3700 +/- 356 pg of angiotensin I h-1 ml-1) and creatinine clearance was lower in the last 12h of salt restriction (arginine vasopressin, 96.1 +/- 6.9; D glucose 116.5 +/- 6.8 ml/min). Renal sodium excretion was unaffected by arginine vasopressin infusion. 5. We conclude that the fall in plasma arginine vasopressin concentration during dietary salt restriction, whilst not affecting renal sodium excretion, may be important in the regulation of plasma sodium concentration, plasma renin activity and glomerular filtration. PMID- 8222514 TI - Natriuretic response to head-out immersion in humans with recent kidney transplants. AB - 1. Recently implanted kidneys may have decreased flexibility to adjust sodium excretion to volume challenges, since modulation by renal sympathetic nerve activity is absent. To examine this hypothesis, we studied the natriuretic response to head-out water immersion in eight patients with well-functioning renal allografts of 37 days (range 24-56 days), at a time when renal re innervation has probably not occurred. 2. By the third hour of head-out water immersion, sodium excretion had increased equally in the patients (from 120 +/- 21 to 204 +/- 37 mumol/min) and in eight healthy control subjects (from 105 +/- 9 to 191 +/- 19 mumol/min). 3. Glomerular filtration rate was 60 +/- 6 ml/min in the patients and 113 +/- 6 ml/min in the control subjects, and did not change upon head-out water immersion. Estimated renal plasma flow increased upon head out water immersion in the control group but not in the patients. Blood pressure decreased similarly in both groups. The renal vascular resistance, calculated from these data, decreased in response to head-out water immersion in the control subjects but not in the renal transplant patients. 4. Head-out water immersion suppressed plasma renin activity only in the normal group, whereas the plasma aldosterone level was suppressed in both groups. The natriuretic response in patients was associated with about 3-fold elevated plasma levels of atrial natriuretic peptide. 5. Since renal re-innervation at 37 days after transplantation is very unlikely, these data suggest that inact renal innervation is not mandatory for a normal natriuretic response to head-out water immersion in humans.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222515 TI - Determinants of the renal clearance of exogenous lithium in a large sample of a white male working population. AB - 1. The associations between the renal clearance of ingested lithium (used as a marker of renal proximal tubular sodium handling) and a number of biological variables have been investigated in an unselected sample of 592 healthy and untreated white men (aged 21-68 years) under their usual living conditions and drawn from a population at work. 2. Renal excretion of lithium was expressed both as clearance of lithium and as fractional excretion of lithium, i.e. normalized for unit of glomerular filtrate. 3. Clearance of lithium was positively associated with a number of anthropometric variables such as weight (r = 0.215; P < 0.001), height (r = 0.212; P < 0.001), body mass index (r = 0.122; P < 0.001) and body surface area (r = 0.244; P < 0.001). However, when expressed as fractional excretion of lithium many of these associations were lost or tended to be negative (weight, r = -0.107, P < 0.01; body mass index, r = -0.119, P < 0.01), greatly depending on the strong relationship between body size and/or mass and glomerular filtration rate. The associations between clearance of lithium and other renal variables reflected, in part, a spurious association mediated by the common relationship with glomerular filtration rate. However, when expressed as fractional excretion of lithium, only some associations were apparent (fractional excretion of sodium, r = 0.256, P < 0.001; fractional excretion of uric acid, r = 0.336; P < 0.001), probably indicating some plausible biological relationships. These results were confirmed by analysis of co-variance by quintiles of clearance of lithium and fractional excretion of lithium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222516 TI - Effect of noradrenaline on renal sodium and water handling in euhydrated and overhydrated man. AB - 1. The renal effects of incremental doses of intravenously infused noradrenaline were evaluated in normal subjects during two different water loads, 5 ml/kg (n = 6) and 20 ml/kg (n = 9), producing conditions of euhydration and overhydration, respectively. 2. Noradrenaline infusion rates ranged from 0.015 to 0.075 microgram min-1 kg-1. In the euhydrated subjects, noradrenaline caused a dose dependent fall in urinary sodium excretion and an increase in urinary flow rate. During overhydration similar doses of noradrenaline caused a fall in urinary sodium excretion but a decrease in urinary flow rate. 3. Although there was no detectable change in glomerular filtration rate, a dose-dependent fall in effective renal plasma flow was observed in both hydration states during noradrenaline infusion. 4. Noradrenaline infusion was associated with a dose dependent increase in proximal tubular sodium reabsorption as assessed by the lithium clearance method. Fractional reabsorption of sodium by the distal nephron was, however, unchanged by noradrenaline in both hydration states. 5. Plasma vasopressin concentration was unchanged by noradrenaline in euhydrated subjects. The renin-angiotensin-aldosterone axis was stimulated by noradrenaline in both euhydrated and overhydrated subjects. 6. Thus we conclude that plasma circulating noradrenaline has a dose-dependent antinatriuretic effect in man. The antinatriuretic effect of noradrenaline is mediated mainly at the proximal tubule in man. We have also shown that during overhydration, noradrenaline decreased urinary flow rate. In contrast, in euhydrated subjects, noradrenaline increased urinary flow rate with no accompanying changes in plasma vasopressin concentration, which suggests a direct effect of noradrenaline on the renal tubular permeability to water. PMID- 8222517 TI - Can ultrafiltration occur with a hypo-osmolar solution in peritoneal dialysis?: The role for 'colloid' osmosis. AB - 1. In peritoneal dialysis the removal of excess body water (ultrafiltration) is traditionally achieved by means of dialysis solution made hypertonic to plasma by the addition of an osmotic agent. In vitro, the osmotic flow may be directed against the osmolality gradient by using a hypo-osmolar solution, but this phenomenon has not previously been applied to clinical peritoneal dialysis. 2. The ultrafiltration performances of hypo-osmolar dialysis solutions containing a high-molecular-weight glucose polymer (weight average molecular weight 22,000), isolated by fractionation of hydrolysed corn starch, were compared with those of hypertonic glucose solutions over a 12 h exchange in 11 patients well established on continuous ambulatory peritoneal dialysis. 3. Five per cent (272 +/- 1.1 mosmol/kg) and 7.5% (277 +/- 2.0 mosmol/kg) glucose polymer solutions produced net ultrafiltration of 243 +/- 53 and 526 +/- 59 ml that were significantly greater than the ultrafiltration of -48 +/- 96 and 223 +/- 84 ml associated with 1.36% (339 +/- 1.9 mosmol/kg) and 2.27% (393 +/- 3.2 mosmol/kg) glucose solutions, respectively. The net ultrafiltration with 10% glucose polymer (284 +/ 2.0 mosmol/kg) and 3.86% glucose (482 +/- 1.6 mosmol/kg) solutions were similar (699 +/- 48 versus 708 +/- 82 ml). 4. The transperitoneal absorption of glucose polymer was substantially lower than that of glucose solutions as was the potential calorie load per millilitre of ultrafiltrate. 5. The addition of 0.35% glucose (molecular weight 180) to 7.5% glucose polymer solution raised the dialysate osmolality to an iso-osmolar level (299 +/- 0.8 mosmol/kg) and produced ultrafiltration which was 29% greater than with 7.5% glucose polymer solution alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222518 TI - Oral mesalamine formulation to be marketed. PMID- 8222519 TI - Dimethyl sulfoxide for treating anthracycline extravasation. PMID- 8222520 TI - Atovaquone: a new antipneumocystis agent. AB - The mechanism of action, pharmacokinetics and pharmacodynamics, clinical efficacy, adverse effects, and dosage of atovaquone in the management of mild to moderate Pneumocystis carinii pneumonia (PCP) are reviewed. Atovaquone has a novel mechanism of action that has been hypothesized to result in microbicidal rather than microbistatic activity against Pneumocystis carinii. Absorption of the drug is significantly enhanced by the presence of food, particularly food with a high fat content. In comparative trials, atovaquone was slightly less effective than trimethoprim-sulfamethoxazole and as effective as pentamidine isethionate in treating mild to moderate PCP. Atovaquone is associated with a lower incidence of treatment-limiting adverse reactions than are trimethoprim sulfamethoxazole and pentamidine isethionate. The most commonly occurring adverse effect in patients receiving atovaquone is rash, and the drug does not appear to cause bone marrow suppression. The FDA-approved dosage regimen for atovaquone in treating mild to moderate PCP is 750 mg (three 250-mg tablets) administered orally three times daily with food for 21 days. Atovaquone may be considered a first-line treatment for patients with the acquired immunodeficiency syndrome who have mild to moderate PCP and have demonstrated an intolerance to trimethoprim sulfamethoxazole. PMID- 8222521 TI - Polymyalgia rheumatica. AB - The epidemiology, relationship to giant cell arteritis (GCA), pathogenesis, pathology, clinical and laboratory features, differential diagnosis, and treatment of polymyalgia rheumatica (PMR) are reviewed. Patients with PMR are usually over 50 years of age, white, and female. There is an association between GCA and PMR that has important implications because of the risk of blindness and other severe vascular complications in patients with GCA. The causes of PMR and GCA are unknown, although the immune system is implicated in the pathogenesis of these diseases. PMR is characterized by muscle pain and stiffness in the shoulders and hips. The principal laboratory finding is an elevated erythrocyte sedimentation rate. The differential diagnosis of PMR includes a number of diseases that cause symmetrical arthritis. It may be particularly difficult to distinguish between PMR and GCA because patients with GCA usually have symptoms associated with PMR. Nonsteroidal anti-inflammatory agents may be effective in mild cases of PMR. However, corticosteroids, usually prednisone or prednisolone, are the class of drugs most widely used to treat PMR. They are effective in relieving the pain and reversing the abnormal laboratory values in most patients; responses can be apparent in 24-48 hours. Steroid-sparing agents such as methotrexate, dapsone, and azathioprine have no established role at present. Patients taking corticosteroids for PMR should be monitored for the occurrence of GCA and development of adverse effects associated with drug therapy. Corticosteroids are effective in treating PMR. Although patients with PMR must be monitored for the development of GCA, the prognosis for these patients is excellent. PMID- 8222522 TI - Drug-induced photosensitivity. AB - Characteristics and diagnosis of photosensitivity are discussed, and drugs available in the United States that cause photosensitivity are identified. In phototoxic reactions, the drug absorbs energy from ultraviolet A (UVA) light and releases it into the skin, causing cellular damage. In photoallergic reactions, light may cause a structural change in a drug so that it acts as a hapten, possibly by binding to proteins in the skin. Once a hapten-protein complex is formed, Langerhans' cells residing in the epidermis can present the antigen to immunocompetent cells, causing hypersensitivity. Phototoxicity is much more common than photoallergy. Drugs that can cause phototoxic reactions include amiodarone, quinolones, and tetracyclines. Drugs that have been associated with photoallergic reactions include thiazides and benzocaine. Pharmacists should be aware of drugs that can cause photosensitivity and should counsel patients taking these drugs to avoid excessive exposure to sunlight. PMID- 8222523 TI - Magnesium in the treatment of acute myocardial infarction. AB - The mechanisms of magnesium action and the possible benefits of its use in treating acute myocardial infarction are reviewed. Magnesium is an essential cofactor in more than 300 enzymatic reactions, including those responsible for the production, storage, and use of energy. It influences impulse generation and action potential propagation of the cardiac and pacemaker cells and affects muscular contraction within the myocardium and arterial smooth muscle. Magnesium has been used successfully for the treatment of arrhythmias and has been shown to produce hemodynamic changes including suppression of vasospasm and reduction of vascular resistance. In clinical trials to assess the usefulness of intravenous magnesium in the treatment of acute myocardial infarction, several beneficial effects were found, including a smaller mean infarction size, a reduction in the occurrence of supraventricular tachycardia, fewer occurrences of serious ventricular arrhythmias, and a lower incidence of early mortality. A bolus dose of magnesium followed by a prolonged infusion to maintain elevated serum magnesium levels appears necessary to obtain the beneficial effects. Studies indicate that magnesium may reduce the incidence of early mortality after acute myocardial infarction. The mechanism of action is still unclear, but it may be a direct cardioprotective effect. PMID- 8222524 TI - Accuracy of a first-order model for estimating initial heparin dosage. AB - The accuracy of a first-order pharmacokinetic model for determining initial heparin infusion rates was studied, and factors that could affect the accuracy of the method were investigated. Patients who received an i.v. infusion of heparin for at least 24 hours for treatment of deep-vein thrombosis, pulmonary embolism (PE), or myocardial infarction were identified by retrospective chart review. A therapeutic dosage of heparin was defined by an activated partial thromboplastin time of 45-75 seconds. Heparin dosages were calculated by using estimated blood volume as the heparin volume of distribution, a desired steady-state heparin concentration of 0.30 units/mL, and an elimination rate constant of 0.832 hr-1. The difference between the calculated dosage and the actual therapeutic dosage was calculated. The differences for various patient subgroups were compared, and the estimated dosages were regressed against the actual dosages to determine their predictive value. Data for 49 patients were analyzed. The mean +/- S.E. difference between the actual and calculated dosages was 29.2 +/- 37.1 units/hr. No significant differences were evident according to sex or indication for therapy. Smokers and nonsmokers differed, as did obese and lean patients. The equation appeared to be more accurate in nonsmokers than smokers. The addition of 200 units/hr to the calculated dosage for patients with PE resulted in minor improvement in the predictive capacity of the equation. Moderate agreement was observed between the actual and calculated heparin dosages in non-smokers of various body weights. PMID- 8222525 TI - Effect of peritonitis on plasma and dialysate alpha 1-acid glycoprotein concentrations in peritoneal dialysis patients. AB - The effect of peritonitis on plasma and dialysate alpha 1-acid glycoprotein (AAG) concentrations was determined. Plasma and dialysate samples were obtained at the onset of infection and one month after treatment from 10 peritoneal dialysis patients with peritonitis. Plasma and dialysate samples were also obtained from 10 noninfected matched controls. Sampling was repeated after a minimum of one month. Samples were assayed for AAG by radial immunodiffusion. The mean +/- S.D. plasma AAG concentrations for patients with peritonitis and for control patients were 152.4 +/- 30.9 mg/dL and 146.6 +/- 45.0 mg/dL, respectively (p > 0.05). The dialysate AAG concentrations for nine control patients were below the limit of detection. The mean +/- S.D. dialysate concentration for the nine infected patients with detectable AAG concentrations was 15.4 +/- 9.5 mg/dL. After successful antimicrobial therapy, dialysate AAG concentrations declined. There was no obvious correlation between dialysate white blood cell count and dialysate AAG concentration during peritonitis. Peritonitis increased dialysate AAG concentrations but had no effect on plasma AAG concentrations. PMID- 8222526 TI - Clearance of N-acetyl-L-tyrosine in infants receiving a pediatric amino acid solution. PMID- 8222527 TI - Encephalopathy associated with muromonab-CD3. PMID- 8222528 TI - Seizure associated with ondansetron. PMID- 8222529 TI - Gentamicin pharmacokinetics in term newborn twins. PMID- 8222530 TI - Criteria for use of lithium in adult inpatients and outpatients. PMID- 8222531 TI - Simulation of ECG from two pairs of action potentials. AB - A simple model for simulation of ECG is presented with the purpose to mimic some common ECG configurations and to generate a hypothesis regarding their electrophysiological background. Action potentials (AP) were simulated on a personal computer from ion currents as described previously (Wohlfart & Arlock, 1993). The difference between two APs of a pair was used to create an electrogram (EG). The subendocardial AP of the pair was triggered by means of a simulated current injection and the subepicardial Ap was triggered from the first AP due to electric coupling within the pair. The subendocardial APs were of longer duration than the epicardial AP because of differences in background currents. A second pair of APs representing another ventricular site was simulated in an analogous way and this pair was activated somewhat later in time. ECG was calculated as the difference between the two EGs. Right-bundle branch block could be imitated by reducing the coupling between the APs representing the right ventricle. Left bundle branch block was generated in an analogous way. ECG in acute myocardial infarction was created after making one of the epicardial APs ischaemic in appearance (reduced amplitude, short duration). Status-post infarction ECG (Q wave and negative T) was produced by reducing the influence from the EG of the infarcted area. Negative T and increased R-wave as in hypertrophy could also be reproduced. Down sloping ST-segment and negative T as in subendocardial ischaemia was also possible to imitate. The simulations showed that biphasic T-waves or T and U-waves follows when the two EGs are separated properly in time. PMID- 8222532 TI - Fatigue in the shoulder muscles during static work at two different torque levels. AB - The present study aimed to investigate whether and when a shift in the mean power frequency (MPF) of the electromyogram (EMG) occurs at low torque levels during a maximum endurance test of three shoulder flexors. Twelve clinically healthy women performed two endurance tests of the shoulder flexors (at 50% MVC--the high torque level, and at approximately 18% MVC--the low torque level) until maximum exhaustion. Surface EMG were obtained and MPF and signal amplitude (RMS) were computed for the trapezius descendens, the anterior part of the deltoid and the infraspinatus. The subjects also rated the perception of fatigue in the shoulder muscles throughout the two tests using a 10-graded scale. A significantly higher degree of perceived fatigue was found at the low rather than at the high torque level. In contrast significantly lower MPF were found at the end of the endurance times in the three muscles at the high torque level when compared to the low torque level. At the low torque level MPF of the trapezius was constant throughout the test. In the deltoid the most prominent decrease occurred during the initial 30-40% of the endurance time at the low torque level. It is suggested that the MPF shift mainly reflects peripheral fatigue of the type-2 fibres. The results of the present study question the use of the MPF shift to monitor peripheral fatigue in the fibres active (mainly type-1) at low torque levels. PMID- 8222533 TI - Relationships between exercise ECG and angiocardiographic indices of coronary insufficiency and myocardial fibrosis in coronary heart disease. AB - Patients with coronary heart disease were examined with exercise ECG and angiocardiography. Maximum work capacity expressed as a percentage of the predicted normal exercise tolerance (Wmax%) was significantly associated with the angiocardiographic score of the myocardial mass subserved by obstructed coronary arteries (MCOS). Variables related to myocardial fibrosis (MF) such as post infarction ECG signs, the left ventricular wall motion score (LVMS) and the ejection fraction of the left ventricle (LVEF) correlated significantly as did variables related to reversible myocardial ischaemia or coronary insufficiency (CI), such as ST depression during exercise (STdepr), ST/W and ST/HR indices, effort angina (EA/W) index, the extent of collaterals (CollS), and 'MCOS-LVMS'. MF variables correlated weakly with CI variables. Wmax% covariated with the variables related to both CI and MF, and most closely with MCOS. Discrepancies between results of exercise ECG and angiocardiography have to some extent been overcome by comparing appropriate parameters. PMID- 8222534 TI - About mechanisms of prostaglandin E1 induced deterioration of pulmonary gas exchange in COPD patients. AB - Prostaglandin E1 (PGE1) has been reported to attenuate COPD-related pulmonary hypertension and to slightly lower the arterial oxygen tension (PaO2). In order to infer the involved mechanisms, the effects of intravenous infusion of PGE1 on pulmonary haemodynamics, diffusing lung capacity for CO (DLCO), membrane diffusing capacity (Dm), pulmonary capillary blood volume (Vc), physiological shunt (Qps/Qt), arterial blood gases and other lung functional indices were evaluated in 20 COPD patients with pulmonary hypertension and, excluding right catheterization, in 14 control subjects. The examines were studied at baseline and during infusion of 20-30 ng kg-1 min PGE1 or placebo. In control subjects PGE1 only caused systemic arterial pressure decrease (-17.8%). In COPD patients, as expected, PGE1 increased cardiac index (16.2%), but decreased systemic arterial pressure (-21.2%), pulmonary arterial pressure (-27.9%), pulmonary vascular resistance (-45.4%) and PaO2 (-10.4%), worsening their hypoxaemia. However, the effect of PGE1 on DLCO was an increase (11.9%), due to an increase in Vc (15.2%) and less markedly in Dm (4.9%). Physiological and anatomical shunts were increased with PGE1 (20.2% and 14.8%) and the overall ventilation/perfusion ratio decreased from 0.89 to 0.79. Decrements in PaO2 correlated with increments in Qps/Qt (r = 0.86). In conclusion, in COPD patients studied, PGE1 increased DLCO, which compensated for the deleterious effect of increased cardiac output on alveolar-capillary gas equilibration. Therefore, worsening of hypoxaemia during PGE1 infusion was related with increased right-to-left shunt and deterioration of ventilation-perfusion relationship. PMID- 8222535 TI - Age-related diminution of the cardiovascular autonomic responses: diagnostic problems in the elderly. AB - To study the usefulness of standard cardiovascular autonomic reflex tests in the elderly, 224 healthy controls and 49 aged diabetic patients were examined. Based on the data obtained from healthy controls, age-related reference values for several autonomic indices were calculated and their usefulness was tested with aged diabetic patients. It was found (1) that in elderly subjects (aged > or = 50 years) the indices based on heart rate differences are more suitable for the assessment the autonomic parasympathetic control than indices based on R/R interval ratios, (2) that the tests (and indices) of choice in the elderly subjects are the Valsalva manoeuvre (Valsalva difference and tachycardia difference) and the active orthostatic test (Max-Rest difference, immediate and later change in systolic blood pressure), (3) that the usefulness of the deep breathing test is limited in the elderly, (4) that such commonly used indices as the Valsalva ratio and the Max/Min ratio in orthostatic test are not useful in the elderly, and (5) that the isometric handgrip test is of little use in the assessment of the autonomic function in the elderly. In conclusion, standard cardiovascular autonomic reflex tests can be used in the assessment of autonomic function to some extent also in the elderly subjects. However, one must bear in mind the limitations in their applicability in that age group. PMID- 8222536 TI - Loss of the nocturnal dip and increased variability of blood pressure in normotensive patients with noninsulin-dependent diabetes mellitus. AB - Eleven normotensive diabetics with noninsulin-dependent diabetes mellitus (NIDDM) (mean age 52.5 +/- SD 8.2 years) and 11 controls (mean age 47.4 +/- SD 8.9 years) had their ambulatory blood pressure and heart rate recorded non-invasively by the Oxford Medilog System in standard hospital conditions. The results were averaged as hourly means of systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial blood pressure (MAP), and heart rate (HR) for the 24-h period and similarly for the 'awake' period (14-16 h) and the 'asleep' period (8-10 h). Hourly means for diabetics and controls showed no differences in blood pressure and heart rate over the 24 h. During sleep, control subjects showed a significant drop in SBP (P < 0.001), DBP (P < 0.001), MAP (P < 0.001) and HR (P < 0.001). However, this nocturnal dip in blood pressure could not be demonstrated in the diabetic group. Blood pressure variability was significantly increased in diabetics compared to controls during waking hours (P < 0.01). These results indicate that in noninsulin-dependent diabetics during sleep there is loss of the nocturnal dip of BP seen in normal subjects, and they have increased BP variability. These may be contributing factors to the development of hypertension and the accelerated target organ damage (TOD) seen in diabetes. PMID- 8222537 TI - Tachycardia and coronary blood flow: non-invasive estimation during Valsalva manoeuvre and exercise. AB - Simultaneous changes in cycle length and coronary blood flow were studied during Valsalva manoeuvre and supine cycloergometer exercise test in 10 male patients (mean age 48 +/- 12 years) who had successfully undergone myocardial revascularization by surgical anastomosis of the left internal mammary artery on the left anterior descending coronary artery. Blood velocity curves in the left internal mammary artery were obtained by a non-invasive continuous-wave Doppler probe at rest, in the last phase of the expiratory effort of the Valsalva manoeuvre and at the maximum load attained during the exercise test. Mean arterial pressure by sphygmomanometer, and cardiac cycle length on the basis of Doppler recording were measured. Mean blood velocity, the length of the blood column entering the coronary bed at each cycle (cardiac cycle times mean velocity), an index of blood cell acceleration (the ratio of mean velocity to cardiac cycle), and an index of coronary resistance (the ratio of mean pressure to mean velocity), were calculated. For approximately the same change in cycle length, coronary resistance decreased in exercise, with an increased mean velocity, but increased in Valsalva, with no changes in mean velocity. The length of the blood column entering the coronary bed at each cycle was unchanged in exercise, with a marked increase in the acceleration index, while it decreased in Valsalva. Therefore, we hypothesize that tachycardia has a limiting effect on sympathetic coronary constriction in Valsalva when cardiac external work is decreased, and an additional vasodilatory effect on coronary bed in exercise when external work is increased. PMID- 8222538 TI - Effect of temperature on finger artery pressure evaluated by volume clamp technique. AB - We examined the effect of temperature on digital arterial blood pressure obtained by continuous beat-to-beat non-invasive monitoring with a volume-clamp technique (Finapres). In 10 normal volunteers and 13 patients with symptoms of vasospasm, digital pressure and brachial artery pressure (cuff method) was simultaneously recorded in control conditions at room temperature, during body cooling, finger heating, and truncal heating. In the control condition digital systolic blood pressure was significantly higher (16.1 +/- 14.2 mmHg) than brachial systolic pressure. The augmentation of digital systolic pressure correlated inversely with finger tip temperature. Diastolic and mean arterial pressure did not differ significantly between the two methods. Body cooling augmented the systolic finger arm gradient while truncal heating and finger heating had the opposite effect. Finger heating reduced systolic augmentation without changing the mean and diastolic blood pressure. Similar changes were also observed in the patients with vasospasm except in one case with a pronounced Raynaud syndrome where digital blood pressure was lower than brachial artery pressure. We conclude that augmentation of finger systolic pressure seems to be dependent on local vasoconstriction of A/V shunts and that finger heating may be a useful procedure to improve the reliability of Finapres readings. PMID- 8222539 TI - Volume blood flow measurements with a transit time flowmeter: an in vivo and in vitro variability and validation study. AB - A transit time flowmeter, Transonic TC101 (Transonic Inc., USA) has been evaluated to study volume flow. Two flow probes, 4 and 6 mm, were used. The experiments were carried out in vivo on the carotid arteries (n = 16) in sheep and in vitro against a pre calibrated rotational pump. Exsanguination measurements, n = 32, were used for the calculation of the error of measurement in vivo. The error of measurement in vitro was calculated on 11 different flow rates from 0 to 150 ml min-1. The variability in vivo was evaluated using data from 10, 30 s measurement periods with a 30 s pause between measurements. The variability in vitro was evaluated at three flow rates (46, 78, and 113 ml min 1). Five 1-min measurements with a 1-min pause between measurements were made. The error of measurement in vivo was 12.05% and in vitro 5.50% (4 mm probe) and 14.90% (6 mm probe). In vivo the variability was between 1.1 and 4.4% and in vitro between 1.5 and 11.2%. The correlation coefficients between the flowmeter on the one hand and exsanguination and the rotational pump on the other were 0.99, 0.99 (4 mm probe) and 0.98 (6 mm probe). It is concluded that the flowmeter has a small variability and error of measurement both in vivo and in vitro. PMID- 8222540 TI - [Use and effect of simvastin in patients with type II diabetes]. AB - Ten non insulin-dependent diabetics in satisfactory metabolic control but with high cholesterol levels were treated with simvastatin. A significant reduction of total cholesterol and LDL-cholesterol was obtained together with an increase in HDL-cholesterol, thus documenting the efficacy of this agent. PMID- 8222541 TI - [Surveillance by skeletal radiography of patients under hemodialysis. Radiographic examination of the vertebral column]. AB - The authors describe damage to the spine observed by x-ray in patients on maintenance hemodialysis. They stress the importance of including x-ray examination of the spine among the routine checks performed in these patients. PMID- 8222542 TI - [Physiopathologic aspects of non-ulcerous dyspepsia. Considerations of a new diagnostic method]. AB - The above study is an analysis of different dyspeptic syndromes outlining their pathophysiology and clinical features. On this basis, the authors suggest a diagnostic strategy, stressing the importance of certain elements such as age at onset, duration and variability of symptoms. This allows to evaluate the indications for endoscopy or biopsy, or to decide that it is sufficient to continue observation while reassuring the patient that the dyspeptic condition is benign. PMID- 8222543 TI - [Alkalizing activity of a calcium-bicarbonate-containing water, evaluated for pH, in patients with gastroesophageal reflux]. AB - Bicarbonate-calcic water Ferrarelle has been administered both in the fasting state and during meals to patients suffering from gastro-esophageal reflux disease submitted to computerized pHmetry. Marked and lasting increase of esophageal and gastric pH was observed with significant differences from the effect of tap water. In addition, patients reported improvement of heart burn and acidity after the administration of the bicarbonate-calcic water. The alkalizing effect of the mineral water employed is therefore fully confirmed. PMID- 8222544 TI - [The epidemiologic problem of glaucoma]. AB - The problem of glaucoma is considered from an epidemiological point of view. The main causes of this pathology are discussed as well as present difficulties of effective prevention. PMID- 8222545 TI - [Diagnostic imaging in the evaluation of non-neoplastic space occupying pathology in the sellar and suprasellar region]. AB - Before the introduction of computed imaging techniques like CT scan and MR, the differential diagnosis of lesions affecting the sellar and suprasellar regions was very difficult, more often impossible. These techniques contributed to ameliorate the typing of pathological tissues. Non neoplastic pathological findings of this region are very different and they can variate between phlogistic processes, vascular lesions, hamartomatous formations or congenital malformations. In this work we describe the characteristic changes of morphology and radiologic aspect in the diagnostic study of this region, using CT scan and MR imaging. PMID- 8222546 TI - [The role of magnetic resonance in the evaluation of pathological processes involving the penis]. AB - The above paper was aimed at defining the usefulness of MRI in the study of pathologies involving the penis. Disorders considered include Peyronie's disease, fibrosis of the penis, priapism, trauma, congenital anomalies, and tumors. In Peyronie's disease and in fibrosis of the penis, MRI permits to demonstrate the abnormal thickening of tunica albuginea and erectile tissue; after administration of gadolinium, it becomes possible in some cases to demonstrate plaques of the albuginea that are not palpable. In priapism, MRI is useful both for identifying the cause and for assessing the time elapsed since blood extravasation. In traumatic and congenital pathologies, MRI is essential for the acquisition of all information on anatomy that may be useful for selecting the most suitable surgical therapy. In neoplastic pathology, MRI has proved useful for accurately assessing the extension of lesions involving the basis of the penis. PMID- 8222547 TI - [The "Castiglione delle Stiviere" project]. PMID- 8222548 TI - [Chemo-antibiotics: individual and social advantages and disadvantages]. PMID- 8222549 TI - [Note on glycosaminoglycans]. AB - Having briefly recalled the chemical structure of glycosamine glycans (GAG) and the therapeutic use of standard heparin at conventional doses (anticoagulant action) and at low dosage (antithrombotic action), the properties and therapeutic employment of low molecular weight heparins (LMWH) are illustrated, and their advantages and disadvantages compared to standard heparin are discussed. Subsequently, the characteristics of heparan sulfates and other GAGs of the vascular endothelium and their therapeutic properties are described. Finally, GAGs found in articular structures are also mentioned. PMID- 8222550 TI - [Nizatidine: a meta-analytical study]. AB - The medline database was applied to search all randomized clinical trials published from 1986 to 1992 concerning the use of Nizatidine, a powerful H2 antagonist, versus other treatments, including placebo, in the treatment of duodenal ulcer disease. Therapeutic efficacy of nizatidine is comparable to that of other H2-antagonists; with the use of meta-analysis, no significant differences were demonstrated, both in acute and long-term treatment of duodenal ulcer between nizatidine and ranitidine, but untoward side effects were less frequent with nizatidine than with ranitidine. PMID- 8222551 TI - [Clinical and pharmacokinetic observations on fluconazole in the management of cryptococcal meningitis]. AB - The above study was intended to evaluate certain pharmacokinetic properties as well as the pharmacological activity of fluconazole in patients with cryptococcal meningitis. The results obtained show satisfactory bioavailability of the drug in the cerebrospinal fluid and marked reduction of the number of fungi found in the liquor. PMID- 8222552 TI - [Effectiveness of ketanserin in the treatment of light-to-moderate arterial hypertension]. AB - The above study was undertaken in order to demonstrate the efficacy of ketanserin in mild to moderate essential arterial hypertension. Twenty-two patients, mean age 49.7 yrs., were studied during 36 months. After a 15-day washout period with clinical and instrumental assessment of basal conditions, treatment with 40 mg ketanserin every 12 hrs. was started. Pressure, blood chemistry and instrumental investigation were carried out systematically and showed the hypotensive action of ketanserin as well as a significant (about 12%) rise in HDL-cholesterol. PMID- 8222553 TI - [Prokinetic drugs in digestive disorders]. AB - In this paper the authors review the main prokinetic drugs analyzing their pharmacological properties, therapeutic indications and adverse effects. In addition, some more recently introduced substances are taken into consideration which, although they were not initially used for digestive tract dyskinesias, can nevertheless be used in certain conditions. From this point of view, calcium blocking agents employed in spastic conditions of the esophagus and the new antiserotoninergic drugs for treatment of anticancer chemotherapy-induced vomiting are of particular interest. PMID- 8222554 TI - [Etiopathogenic and clinical considerations on primary pulmonary hypertension]. AB - Primary pulmonary hypertension is a severe condition of unknown etiology first described by Romberg in 1891. The authors review the literature with particular emphasis on new hypotheses concerning etiopathogenesis, and recent suggestions for management. The former take into account the discovery of endothelium derived relaxing factors and the identification in pulmonary arteries of a polypeptide called endothelin. Further hypotheses contemplate 1) interaction between platelets and endothelium; 2) the role of heparin-inhibiting enzymes; 3) abnormal cellular adhesion. Heart catheterism is apt to exclude other causes of pulmonary hypertension and also permits to assess the severity of the condition and its prognosis, as well as acting as a guide to management. If venous O2 saturation is higher than 63%, calcium blocking agents and anticoagulants should be used, if it is lower than 63% long-term prostaglandin infusion should be given awaiting heart lung transplantation unless this can be done immediately. PMID- 8222555 TI - [Characterization of space occupying masses in the diencephalic-hypothalamic region. MRI vs. TC]. AB - All morphologic aspects of neoplastic and non neoplastic masses, arising from the diencephalic-hypothalamic region are discussed in this paper. We describe the densitometric characteristics on CT scan and the MRI appearance of masses affecting this region, both of benign lesions and malignant neoplastic masses. We describe separately masses arising from within the III ventricle and those of extraventricular origin and all semiotic findings of neuroradiological techniques of computed imaging. PMID- 8222556 TI - Effect of aging on respiratory skeletal muscles. AB - Although it has been recognized that limb skeletal muscles undergo changes during aging, it has been unclear to what extent age-related changes occur in respiratory muscles. This article reviews the structural organization of human skeletal muscles, including those of the respiratory system. Evidence that changes do occur in the structure and function of these muscles in elderly individuals is presented. The potential clinical significance of aging on respiratory muscle strength and endurance is discussed. PMID- 8222557 TI - Immunology of the aging lung. AB - The aging process is associated with multiple deficits in pulmonary immune function. Defense of the airway is impaired in the elderly by decreased mucociliary clearance, alteration in respiratory mechanics and, in some cases, concomitant illnesses that predispose to aspiration. Alveolar defenses can be divided into resident defense mechanisms, inflammatory responses, and specific immune responses. Resident defenses such as macrophage phagocytosis and chemotaxis, although largely intact, may have subtle defects under specific conditions. Inflammatory responses may also be diminished, as evidenced by decreased neutrophil-mediated killing and chemotaxis. Specific immune responses appear to be the most vulnerable to age-associated impairment. Although antigen presentation is well preserved during aging, accessory cell cytokine production may be decreased. T-lymphocyte proliferative responses are markedly decreased, as is the production of and response to intercellular mediators. Age-associated alterations in T cell subpopulations may result in imperfect T cell-B cell interactions leading to expression of abnormal immunoglobulins. These many interacting and compounding impairments may explain the increased susceptibility of the elderly to pulmonary infection and autoimmune diseases. PMID- 8222558 TI - Evaluation of dyspnea in the elderly patient. AB - The symptom of dyspnea in the elderly person should not be considered part of the "normal aging process." Instead, the history, examination, and testing should focus on cardiac disease, respiratory disease, and deconditioning as the most likely causes. Because respiratory sensation is diminished with aging, breathlessness may not develop until a more advanced stage of disease or dysfunction. Clinical measurement of dyspnea is important to assess its severity and to evaluate response to treatment. Specific treatment should be directed toward the pathophysiology of the underlying disease. General strategies for relieving dyspnea include breathing techniques, exercise training and reconditioning, oxygen therapy, improved nutrition, and, in selective cases, psychotropic medication. PMID- 8222559 TI - Sleep in the elderly patient. AB - Sleep problems in the elderly are extraordinarily common. The authors discuss normal sleep, changes in sleep with normal aging, and sleep disorders in the elderly, focusing on sleep-disordered breathing. In addition, nocturnal myoclonus and rapid-eye movement behavior disorder are reviewed. PMID- 8222560 TI - Smoking cessation in the elderly patient. AB - An older smoker tends to be asked less often to quit although he or she has much to gain from achieving a nonsmoking status. Multiple barriers may interfere with attempting to quit including nicotine withdrawal, doubted self-efficacy, other smokers in one's environment, or misperceptions of little gain from quitting. The physician is in a unique position to interact with the older smoker and through careful questioning, outline a quit smoking strategy that has the greatest chance for success. Every older smoker deserves a chance for increased quality as well as quantity in life. PMID- 8222561 TI - Pulmonary rehabilitation in the elderly patient. AB - Pulmonary rehabilitation in the elderly can be a rewarding experience for the patient and the health care provider. Rehabilitation programs are well established as a means of enhancing standard medical therapy, optimizing functional capacity, and restoring a patient to the highest possible level of independent function. The demonstrated benefits of pulmonary rehabilitation include an improved quality of life, increased exercise endurance, improved ability to perform daily tasks, reduced hospitalization, and improved psychosocial adjustment. Participation in a group program helps reduce social isolation, whereas postprogram patient-support groups help to keep patients motivated and positive in their outlook. Through pulmonary rehabilitation the elderly patient learns to accept and overcome the reality of his or her disease rather than dwell on its limitations. An individually tailored comprehensive rehabilitation program can make the difference between a fulfilling life and a life of pulmonary disability. PMID- 8222562 TI - Special considerations in the elderly patient. AB - This article has discussed drug therapy in the elderly and included a discussion of optimal inhaled bronchodilator therapy; issues relating to tracheostomy, a frequent requirement after ventilatory failure in the elderly; oxygen administration, emphasizing its effects on exercise in COPD, and forced oscillation, a new technique that may have utility in the measurement of lung function in the geriatric population. PMID- 8222563 TI - Lung cancer in the elderly patient. AB - One half of all cases of lung cancer occur in patients aged 65 years and older. Although adenocarcinoma is the predominant histologic subtype in the general population, squamous cell carcinoma has remained the most common type in the elderly. Although lung cancer tends to present at a less advanced stage with increasing age, the percentage of elderly patients undergoing treatment tends to decline despite the fact that they can tolerate the aggressive therapy that may enhance survival. PMID- 8222564 TI - Nosocomial pneumonia in the elderly patient. Chronic care facility and hospital considerations. AB - By nature, nosocomial infections in general, and pneumonia specifically, are opportunistic, preying on patients who are already seriously ill. These infections are a major source of morbidity and mortality for these individuals. Among the elderly, pneumonia is more common and more likely to lead to death. The author reviews the epidemiology and risk factors for nosocomial pneumonia among the elderly. PMID- 8222565 TI - Tuberculosis. An epidemic in older adults. AB - The incidence of tuberculosis is disproportionately high in our elderly population. This group appears to be at high risk because many were infected early in life. Additional factors contributing to increased risk in the elderly probably include low clinical suspicion for tuberculosis in this age group, unusual or nonspecific clinical and roentgenographic presentations, difficulties in interpreting the tuberculin skin test, and the presence of associated illnesses that may lead to delay in diagnosis. Failure to diagnose tuberculosis in this population has important public health implications, particularly in nursing homes and long-term care facilities. Tuberculosis is usually easily diagnosed when suspected in elderly patients, and effective therapy is available for prevention and treatment of this disease. The special needs of old patients must be considered when these regimens are applied. Increased awareness of tuberculosis in the elderly is important for the success of strategies that have been developed to eliminate tuberculosis from the United States. As our population continues to age, physicians who treat the elderly must be vigilant in suspecting tuberculosis as a cause of illness in these patients. PMID- 8222566 TI - Infiltrative and interstitial lung disease in the elderly patient. AB - The interstitial lung diseases encompass a wide array of inflammatory and fibrotic processes that produce a significant amount of morbidity in the elderly. Diagnosis usually requires a systematic approach and often requires lung biopsy. Treatment typically involves the use of immunosuppressive dosages of corticosteroids or cytotoxic agents. The use of these medications in the elderly demands meticulous, close follow-up because side-effects can be substantial. PMID- 8222567 TI - Thrombophlebitis and pulmonary embolism in the elderly patient. AB - The incidence of deep venous thrombosis and subsequent pulmonary embolism is increased in the elderly but, because the clinical manifestations may be misleading, the diagnosis is often missed. The clinician must maintain a high index of suspicion and order specific tests for venous thrombosis or pulmonary embolism whenever these diagnoses are considered. Even though the risk of bleeding while anticoagulated is increased in the elderly, treatment with heparin followed by long-term anticoagulation with warfarin is the preferred course of therapy for most patients. Awareness of their increased risk and emphasis on prevention of venous thrombosis may be the most effective means of dealing with this clinical problem in the elderly. PMID- 8222568 TI - Community-acquired pneumonia in the elderly patient. Clinical features, epidemiology, and treatment. AB - The relationship between age and the incidence of and mortality associated with pneumonia is complex. Age itself may not be an independent risk factor, because elderly patients also have a concomitant increase in the number of underlying risk factors for disease. The authors review the clinical features, epidemiology, and treatment of community-acquired pneumonia. PMID- 8222569 TI - Nonresolving and slowly resolving pneumonia. Diagnosis and management in the elderly patient. AB - Pneumonia is both more prevalent in the elderly and more severe. This article reviews the approach to nonresolving or slowly resolving pneumonia in the elderly. Specific attention will be given to the natural history of commonly occurring pneumonias in this age group, related host defense impairments, unusual infectious causes of slowly resolving pneumonia, and noninfectious mimics of pneumonia. PMID- 8222570 TI - Approach to adult respiratory distress syndrome and respiratory failure in elderly patients. AB - Because of physiologic changes associated with aging and the development of chronic illness, the elderly will continue to be at risk for catastrophic illnesses including adult respiratory distress syndrome (ARDS). Current management of elderly patients with ARDS is only supportive and focused on avoiding complications. Truly successful management of these patients requires therapies directed at pathophysiologic initiators and mediators of ARDS. PMID- 8222571 TI - Outcome assessment in elderly patients with critical illness and respiratory failure. AB - Although elderly patients tend to have diminished physiologic reserve, the independent impact of age on outcome from critical illness is controversial. Physiologic status is perhaps a more important measure of a patient's underlying health and anticipated response to critical illness. Age, therefore, should not be used as a sole determinant for intensive care department admission. PMID- 8222572 TI - Ethical considerations and CPR in the elderly patient. AB - Cardiopulmonary resuscitation has become an increasingly common event in the terminal illness of hospitalized seriously ill elderly patients. Elderly patients, however, tend to respond poorly to the resuscitation procedure. The authors analyze the ethical and legal considerations in offering or withholding cardiopulmonary resuscitation. The central position of the physician in decision making is emphasized. PMID- 8222573 TI - Molecular mechanisms of thyroid hormone action. AB - This review summarizes our current understanding of the mechanisms of thyroid hormone action. Topics included are the structures of thyroid hormone response elements (TREs), thyroid hormone receptor (TR)-TRE interactions, and TR association with other proteins to illustrate their importance in modulating both positive and negative transcriptional activation of specific genes. Certain studies focusing on other members of the steroid/thyroid hormone receptor superfamily are covered to provide a current view of how this class of receptors stabilizes transcription initiation complexes. PMID- 8222574 TI - Molecular mechanisms of thyroid hormone action. A physiologic perspective. AB - At present, it appears abundantly clear that thyroid hormone exerts its major action at the nuclear level by regulating the level of mRNAs of specific genes. There are at least three TR isoforms that mediate hormonal effects at the tissue level. Characterization of the functional domains of these receptor isoforms is as yet incomplete, and the possibility that these receptors could have ligand independent functions is a matter under current investigation. TRs are now recognized as members of a large superfamily of transactivating proteins involved in the regulation of gene expression. Recent studies have shown an unexpected degree of complexity in the nature of the association of the T3 receptors and the DNA of target genes. They have vividly pointed out the multiple interactions possible between the T3-receptor complex and other proteins participating in the process of gene regulation. These insights have provided a solid base for understanding differences in the gradation of thyroid hormone effect from one tissue to another. The microdissection of the molecular process that has occurred in the past 20 years has proceeded in part through the application of relatively artificial in vitro systems and assays. Whereas such approaches have undoubtedly reaped rich rewards in pointing out potential or possible mechanisms, they do not define the actual workings in the animal. Additional studies designed to examine at the molecular level the operation in vivo of physiologic networks influenced by thyroid hormones appear as an essential next step in understanding the biology of the hormone system. The application of transgenic models should materially assist such efforts. PMID- 8222575 TI - Resistance to thyroid hormone. AB - Resistance to thyroid hormone encompasses a clinically heterogeneous group of conditions characterized by reduced responses of target tissues to a supply of thyroid hormone that under normal circumstances would be excessive. The syndrome of resistance to thyroid hormone is first suspected when serum thyroid hormone levels are found to be elevated in the presence of nonsuppressed serum thyrotropin (TSH). Clinical confirmation of the diagnosis requires the demonstration of the failure of full replacement doses of thyroid hormone to produce the expected suppressive effect on the secretion of TSH and a failure to induce appropriate responses in peripheral tissues. PMID- 8222576 TI - Thyroid hormone transport proteins. AB - TBG, TTR, and albumin are quantitatively the most important thyroid hormone binding proteins in humans. Only a minute fraction of T3 and T4 circulates unbound, but it is this free hormone that is metabolically active at the tissue level, and, therefore, responsible for thyroid status. Inherited or acquired variations in the concentration or affinity of these proteins may produce substantial changes in serum total thyroid hormone levels but do not affect serum free thyroid hormone concentrations and, therefore, do not influence actual thyroid status. Thus, thyroid hormone-binding protein abnormalities must be suspected when abnormally elevated or diminished total thyroid hormone concentrations are encountered in clinically euthyroid subjects. This is crucial to avoid erroneous and potentially detrimental treatments for hyperthyroidism or hypothyroidism. PMID- 8222577 TI - Analytic measurements of free thyroxine. AB - This article has attempted to clarify two main areas of uncertainty relating to the clinical use of free TH determinations. The first centers on the validity of the free hormone hypothesis. This has been the subject of contentious debate for more than a decade, and final agreement among endocrinologists on this issue has yet to be reached. It is thus important that diagnosticians should not assume the validity of the hypothesis when assessing the analytic validity of free TH assay kits. In short, the reliability of the kits should be judged by analytic criteria (such as their theoretic basis and the results yielded on diluted samples) and not merely on the claim that they yield correct results in certain patient categories. Indeed, the finding that fT4 decreases in pregnancy (albeit to a limited extent) illustrates the danger of assuming the validity of the hypothesis when judging the reliability of fT4 assay kits. The second area of uncertainty relates to the true serum fT4 concentrations present in various clinical conditions, in consequence of doubts regarding the analytical validity of currently available methods. An attempt has been made in this presentation to clarify the physicochemical concepts underlying the more widely-used kits, and some of the reasons for their unreliability, although this also constitutes a contentious area. Errors generated in analogue-based free TH assay kits arise in part in consequence of the use of analogues that fail to conform to the physicochemical theory underlying "unbound analogue" assay, compounding errors resulting from the inclusion in assay reagents of various additives (e.g., albumin) that, inter alia, disturb the equilibrium between free and bound hormone moieties, and which also distort the effects of binding competitors. Regrettably different manufacturers have, in practice, relied on different analogues and/or reagent additives, buffers, blockers, and so forth, and different operating temperatures. Indeed different kits from the same manufacturer frequently differ in their formulation (see, for example, footnote on page 618), and yield widely differing results in certain sera. Also manufacturers may modify kit components from time to time without disclosure. Thus, though many analogue-based kits may arguably be diagnostically useful in well-defined clinical situations, total reliance cannot be placed in the results they yield in more complicated circumstances (such as those characterizing NTI), which may be diagnostically misleading.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8222578 TI - Automation of thyroid function testing. AB - This article discusses the automation of thyroid function testing. The needs for automation are included in the introduction. The issues concerning automation of immunoassay are discussed, including homogeneous or heterogeneous immunoassay, competitive or immunometric assay, reagent stability, data management, sample management, signal detection, and the disadvantages of automation. Individual immunoassay systems are summarized with a table outlining the key features. Both technical and clinical performance of automated thyroid function testing are described. The article ends with a discussion of the future trends of automated immunoassay testing. PMID- 8222579 TI - Neonatal screening for congenital hypothyroidism. PMID- 8222580 TI - Clinical application of free thyroxine determinations. AB - The clinical diagnosis and management of thyroid hormone excess and deficiency are dependent on accurate laboratory measurements and the interpretation of serum free T4 and TSH values. A variety of free T4 methods are available that perform well in otherwise healthy patients with hypothyroidism or hyperthyroidism and in euthyroid subjects with mild alterations of T4 binding to serum-carrier proteins. In contrast, only free T4 values by direct equilibrium dialysis, a method that is available in larger clinical laboratories, and ultrafiltration of undiluted sera, which is a research method, provide appropriate free T4 values in the majority of patients with significant alterations of serum T4 binding, including severe nonthyroidal illnesses. In patients with a normal pituitary-thyroid hormone axis, an inverse log10-linear relationship exists between serum TSH and free T4 levels; a decreased direct equilibrium dialysis free T4 value with an elevated TSH level confirms the diagnosis of primary hypothyroidism, and an increased free T4 value with a TSH level less than 0.01 mU/L is consistent with nonpituitary hyperthyroidism. When this relationship is altered, as in nonthyroidal illnesses, TSH secreting tumors and thyroid hormone resistant states, a direct equilibrium dialysis free T4 level plus a third-generation TSH assay is the most sensitive and specific approach to diagnose thyroid hormone excess or deficiency. Use of other free T4 methods in patients with significant alterations of serum T4 binding results in a variably increased frequency of false-positive values, which require additional testing to define the thyroid hormone status. PMID- 8222581 TI - Role of thyrotropin measurements in the diagnosis and management of thyroid disease. AB - This article reviews the role of thyrotropin assays in clinical medicine and illustrates the advantages and limitations of assays that can measure subnormal concentrations. Analytic considerations that can significantly alter the performance of these assays are outlined. The concept of assay generations is defined, and the potential roles of third and fourth generation assays are postulated. In addition, a diagnostic strategy is presented for using TSH and other procedures in the diagnosis and management of thyroid disease. PMID- 8222582 TI - Autoimmune thyroid disease. AB - A considerable proportion of the thyroid diseases is due to aberrant immune reactions toward thyroid antigens. Autoreactivity is considered to be a normal process controlled by several suppressor mechanisms. Malfunction of these suppressor mechanisms may result in autoimmune disease. Mechanisms of self tolerance, defects in autoimmune disease, thyroid autoantibodies, thyroid autoreactive T cells, several thyroid autoimmune diseases, and the role of iodine in thyroid autoimmune reactivity are addressed. PMID- 8222583 TI - Fine-needle aspiration cytology of the thyroid. A 12-year experience with 11,000 biopsies. AB - The primary purpose of FNA biopsy is to identify benign nodules and avoid unnecessary surgery. Numerous reports confirm that the introduction of FNA reduces thyroid operations by 25% and increases the yield of carcinoma from 14% to at least 30%. With an improvement in surgical selectivity, the yield of carcinoma has significantly increased, and the impact of FNA on thyroid practice has been substantial. The economic impact of FNA biopsy is also considerable and makes this test a useful and cost-effective procedure. Our approach to a patient with nodular thyroid disease is outlined in Figure 1. This scheme rests on thyroid cytology, and FNA biopsy is used as the first diagnostic test. When an experienced clinician performs the aspiration and an experienced cytopathologist reviews the slides, the accuracy of the technique is better than 90%. In agreement with recent FNA series, we believe that FNA biopsy is a safe and reliable procedure and that complications are extremely rare. On the basis of our experience, its continued use as the first step in the diagnostic management of thyroid nodules seems justified. PMID- 8222584 TI - Thyroid imaging techniques. AB - Nuclear medicine, ultrasound, CT, and MRI are imaging methods that can be used to evaluate the thyroid gland. All these techniques give structural information about the thyroid gland and show the location and size of thyroid nodules. Nuclear medicine scanning also adds functional information about nodules. In many practices, however, FNA has supplanted imaging methods as the primary method of thyroid nodule evaluation because it is safe, inexpensive, and results in a better selection of patients for operation. Imaging studies are very useful in the setting of recurrent thyroid cancer. Ultrasound is extremely sensitive in the detection of recurrent malignancy in regional cervical lymph nodes and as a guide in performing a biopsy of these nodes. CT is very useful in identifying distant metastases in the chest and abdomen. Nuclear medicine scanning can detect functioning distant metastases when the metastases are from differentiated thyroid cancers. MRI can be used to evaluate the possibility of recurrent thyroid cancer; however, because of its relatively high cost, it is used less frequently than other imaging methods. PMID- 8222585 TI - Thyroid cancer diagnosis and management. AB - This article provides an overview of the classification of thyroid carcinoma into well-differentiated forms (papillary and follicular) as well as the rare Hurthle cell, medullary, and anaplastic forms. The characteristics, relative frequency, and mortality rates of each form of thyroid cancer are summarized. The roles of history, physical examination, blood tests, such as thyroid function test, thyroglobulin (htg), calcitonin and CEA, radionuclide and ultrasound imaging, and fine-needle aspirates are outlined for the diagnosis and management of thyroid cancer. An algorithmic diagram of the monitoring procedures for long-term management of various forms of thyroid cancer is presented. PMID- 8222586 TI - Federal regulation of clinical laboratories and the Clinical Laboratory Improvement Amendments of 1988--Part I. AB - In this first part of a two-part article, we have summarized some of the pertinent features of CLIA 1988 with review of the complexity model, waived testing, personnel standards, proficiency testing and quality control requirements, and patient test management. In addition to a survey of other Federal regulations applicable to clinical laboratories, we have briefly reviewed events subsequent to the publication of the Final Rule in February of 1992, specifically the recommendations of CLIAC and the Technical Corrections published on January 19, 1993. The second part of this paper will discuss sanctions and enforcement; how laboratory inspections are likely to be performed, accreditation of private organizations, state exemptions, cytology and other specialty regulations, and the evolving interaction between CLIAC, FDA, CDC, DHHS, and the regulated laboratory community. PMID- 8222587 TI - Use of ultrasound in obstetrics. PMID- 8222588 TI - Supporting the family of the newborn with Down syndrome. PMID- 8222589 TI - Exercise-induced menstrual irregularities. AB - Exercise of sufficient rigor, particularly when coupled by weight loss and dietary restriction, is capable of producing reversible disturbances of reproductive function in many otherwise healthy women. However, it is clear that there are many health benefits for women who exercise regularly and in moderation. These include improved cardiovascular status, increased bone mineral content, improvement of dysmenorrhea and premenstrual syndrome symptoms, and general stress reduction. In addition, the inclusion of young women in high school and college athletics has provided many new opportunities. With these new opportunities, however, have come new challenges for physicians in assisting these women in maintaining optimal health. PMID- 8222590 TI - How old is the fetus? PMID- 8222591 TI - Calcium homeostasis in normotensive and hypertensive pregnancy. PMID- 8222592 TI - Women's health problems. PMID- 8222593 TI - Inflammatory bowel disease in the pregnant woman. AB - Although the woman with IBD possesses a greater potential for a complicated pregnancy, the majority of patients will experience an uneventful normal pregnancy. It is important to educate the young patient with IBD when she is planning a family. Conception at a time when IBD is quiescent offers the greatest likelihood of an uncomplicated pregnancy. Physicians must recognize and inform their patients that most medications that are necessary to suppress the disease should be continued throughout pregnancy. Although generalities can be made regarding the management of pregnant women with IBD, the individual patient may need specifically tailored therapy for her individual case. PMID- 8222594 TI - Differentiating between natural progesterone and synthetic progestogens: clinical implications for premenstrual syndrome management. PMID- 8222595 TI - The spectrum of osteitis pubis. PMID- 8222596 TI - Secular trends of caries prevalence in 6- and 12-year-old Dutch children. AB - Recently two papers have been published dealing with possible changes in the downward trend of caries experience among Dutch schoolchildren in the late 1980s. To answer the question if the decline of caries has come to an end, a meta analysis was carried out on epidemiological data collected in 5-6 and 11-12-yr old children between 1980 and 1990. From results of the present analysis it appears that after 1983 a halt in the decline of caries prevalence among 6-yr olds occurred, whereas among 12-yr-olds, the earlier decrease in mean DMFS scores continued in the period 1980-89. PMID- 8222597 TI - A retrospective analysis of the costs associated with the treatment of nursing caries in a remote Canadian aboriginal preschool population. AB - Nursing caries is a specific form of rampant dental caries affecting the majority of preschool aboriginal children who live in the Province of Manitoba, Canada. Since the majority of these individuals live in remote regions of the province access to dental treatment is difficult, resulting in long delays in the provision of treatment and, most likely, significant morbidity associated with dental pain and oral infection. Travel to distant centres for treatment under general anesthesia by pediatric dentists has become the usual method by which treatment is provided to the majority of affected children. We believed that this was an expensive method of providing these necessary services and our purpose was to document all costs associated with the treatment of nursing caries in this population. We analyzed the records of 884 children who were treated for nursing caries between 1980 and 1988 in Manitoba and collected data for costs in the following categories: travel, lodging, medical, dental, hospital and nursing. Our results show that the remote band groups had significantly higher costs (P < 0.001) than groups which were located closer to treatment centres. The costs which accounted primarily for this significant difference were travel and medical costs associated with hospitalization and the administration of general anesthesia. Our results support the need for the redeployment of resources on the basis of regional need and the development of community-based preventive programs and treatment programs which will significantly reduce the incidence of nursing caries in preschool Canadian aboriginal children. PMID- 8222598 TI - Factors influencing the effectiveness of sealants--a meta-analysis. AB - Meta-analysis of published studies was applied to analyze the effectiveness of fissure sealants in preventing dental caries. Studies were identified by a MEDLINE search, supplemented by a hand search of the references in the articles recovered. All articles were assessed on the basis of quality, and were combined with the Mantel-Haenszel method. The prevented fraction in the exposed population (PF) was estimated using pooled relative risk. Heterogeneity of the effect and publication bias were also analyzed. Twenty-four studies were included in the meta-analysis. The overall effectiveness of autopolymerized fissure sealants was 71.36% (95% confidence interval 69.69-72.94%). Effectiveness in preventing caries decreased with time, and increased when drinking water was fluoridated (82.69% vs 72.28%). Heterogeneity was significant in most cases, and was further studied with multiple linear regression analysis. It is concluded that autopolymerizing sealants should be used. More research is needed to compare the effectiveness of dental hygienists and dentists in applying sealants. PMID- 8222599 TI - Applicability of Receiver Operating Characteristic (ROC) analysis on discrete caries depth ratings. AB - Accuracy of diagnostic tests for caries detection is frequently evaluated by sensitivity and specificity values obtained from data acquired on an ordinal rating scale which classifies lesions according to the depth in enamel and dentin. The interpretation of these parameters may, however, be difficult. Receiver Operating Characteristic (ROC) analysis, on the other hand, yields an expression for overall diagnostic accuracy in a single parameter, facilitating a comparison of performance of several diagnostic tests. The basis for applying ROC analysis has been that data were collected on a confidence (likelihood) scale. The aim of this preliminary study was to investigate whether the application of ROC analysis would yield as valid parameters for diagnostic accuracy in assessment of approximal caries on a depth rating scale as on a likelihood rating scale. The results demonstrated that the accuracy of observer performance was not influenced by the type of rating scale employed, justifying the application of ROC analysis on discrete caries depth ratings. PMID- 8222600 TI - Assessment of reports by dentists of their restorative treatment thresholds. AB - Questionnaires have been used to determine the point when dentists think a filling ought to be placed (restorative treatment threshold). This study assesses the method. 211 general dental practitioners were interviewed and asked to identify the point at which they would instigate restorative treatment in a 12-yr old patient from a list of descriptions of lesions affecting buccal/lingual, occlusal and approximal surfaces. They were also asked to judge the likely depth and lateral spread of the lesion they chose and to identify a photograph resembling it (buccal/lingual and occlusal surfaces) or a line drawing and description of its expected surface appearance (approximal surfaces). The Pearson correlation coefficients between reported treatment threshold and the characteristics expected to be associated with them were poor. This suggests that there may be considerable variation in opinion between dentists about the underlying condition of carious lesions of similar surface appearance. There was no evidence that the treatment opinions of these dentists have changed markedly between 1987 and early 1991. PMID- 8222601 TI - Epidemiological aspects of lip cancer in Scotland. AB - The purpose of this paper is to present the descriptive epidemiology of lip cancer in Scotland. The incidence and mortality of lip cancer are found to be higher in men than in women and to have fallen remarkably among men. This decline has been the result of cohort effects indicating that the risk of lip cancer is decreasing among successively younger people. PMID- 8222602 TI - Configural frequency analysis for exploring bacterial sets in periodontal health and disease. AB - Periodontal research has been concerned with identifying bacterial sets in oral health and disease. Configural frequency analysis (CFA) is a taxonomical method used to test the statistical significance of patterns of elementary cell frequencies in cross-tabulated data, and may therefore be used to evaluate interactions among occurrences and non-occurrences of periodontal microorganisms. CFA patterns may exist as configural types or anti-types depending on whether observed cell frequencies are greater or less than expected frequencies, respectively, based on table marginals. CFA was applied to two published data sets. In the first, the occurrence of Bacteroides (Porphyromonas) gingivalis (Bg) and Treponema denticola (Td) was studied in military subjects. Analysis of individual cells of the resultant two-by-two contingency table revealed the existence of a single anti-type (Td in the absence of Bg) suggesting a sequential phenomenon. The second, more complex, data set was based on reported occurrence of Actinobacillus actinomycetemcomitans (Aa), Bg, and Bacteroides (Prevotella) intermedius (Bi) in sites with and without progressive periodontitis. CFA based on all data identified three statistically significant configural types and two anti-types. However, a somewhat different configurational picture emerged when disease progression status was considered a stratification variable for separate analysis. All these analyses, which were undertaken for expository purposes, assumed independence of observations. Methodological procedures are suggested for CFA when observations are not independent. PMID- 8222603 TI - Mutans streptococci and caries prevalence in preschool children. AB - Numerous studies have reported a correlation between mutants streptococci levels and dental caries. The aim of this study was to assess the relationship between salivary mutans streptococci levels and caries in preschool children of low socioeconomic status. A total of 462 Head Start children, mean age 3.8 yr (range 2.0-5.3 yr), were examined by the modified method of Radike. Saliva samples from 458 of these children were collected with tongue blades and impressed onto mutants streptococci selective agar. Children's mutants streptococci levels were categorized as low (0 CFU), moderate (1-50 CFU) or high (> 50 CFU), and the mean dmfs was 0.40, 1.92 and 4.88, respectively. All study groups (Black, Hispanic and White) had infection rates of approximately 83%; however, 39.1% of Black children had high mutans streptococci levels compared with 28.4% of White children. Pit/fissure caries was the most prevalent disease type in children with moderate or high mutants streptococci levels, although White children in the high group had significantly less of this pattern than Blacks and Hispanics. Sensitivity, specificity, and positive and negative predictive values for the high mutants streptococci group were 91.3%, 57.5%, 69.3% and 86.3%. Results from this study indicate that differences between Black, Hispanic and White preschool children may influence caries activity within populations that have similar mutants streptococci infection levels and socioeconomic backgrounds. PMID- 8222604 TI - Prevalence and characteristics of dental anxiety in Danish adults. AB - Prevalence, characteristics and consequences of dental anxiety in a randomly selected sample of 645 Danish adults were explored in telephone interviews. Participation rate was 88%. Demographics, fear of specific procedures, negative dentist contacts, general fear tendency, treatment utilization and perceived oral conditions were explored by level of dental anxiety using a modified Dental Anxiety Scale (DAS). A Seattle fear survey item and a summary item from the Dental Fear Survey (DFS) were also included for fear description comparisons. Correlation between these indices (DAS-DFS: rs = 0.72; DAS-Seattle item: rs = 0.68) aided semantic validation of DAS anxiety intensity levels. Extreme dental anxiety (DAS > or = 15) was found in 4.2% of the sample and 6% reported moderate anxiety (DAS scores 14-12). Bivariate (B) and logistic regression (L) odds ratios (OR) showed that high dental anxiety was associated with gender, education and income, but not with age. Extreme dental anxiety for dentate subjects was characterized by fear of drilling (ORL = 38.7), negative dentist contacts (ORL = 9.3), general fear tendency (ORL = 3.4), avoidance of treatment (ORL = 16.8) and increased oral symptoms (ORB = 4.4). Moderate dental anxiety was also related to drilling (ORL = 22.3), but with less avoidance due to anxiety (ORL = 6.8) compared with low fear subjects. PMID- 8222605 TI - How Swedish dental hygienists apply their training program in the field. AB - The purpose of this study was to evaluate how clinical practice by Swedish dental hygienists was related to type of dental delivery system, period of training, educational institution attended and patient category. Dental hygienists from 14 different dental hygiene schools were represented. Of these schools, 11 are still in operation. A specially designed questionnaire was posted to all dental hygienists in Sweden (n = 1857). A total of 1399 questionnaire (75.3%) were completed and returned, providing data on 15,546 dental appointments. 37.2% of the Swedish dental hygienists are presently working in private practices, 45.8% in the public dental health service and 6.2% in both. Of the patients treated by hygienists, 88.7% were adults: 99.5% in private practice and 78.4% in the public dental health service. 42.0% of all dental hygienists were trained in 1980-84. The mean treatment time per appointment in private practice was 49.7 min and 45 min in the public dental health service. Scaling, root-planning and removal of overhangs took 27 min per visit in private practice and 22 min in the public dental health service. However, there were no significant differences in methods in the two delivery systems with respect to examinations, self-care training, professional mechanical toothcleaning (PMTC), topical fluoride application, or salivary and oral microbiology tests. The adult patient categories were periodontal risk (45.1%), caries risk (9.1%) and hygiene (34.6%). In periodontal risk patients, scaling, root-planning and removal of overhangs took 28 min per appointment and 14 min in caries risk patients. PMID- 8222606 TI - Prevalence of malocclusion in high school students in Japan according to the Dental Aesthetic Index. AB - The Dental Aesthetic Index (DAI) was used to evaluate malocclusion in Japanese high school students (n = 409, ages 15-18 yr). The purpose of this study was to obtain basic data concerning malocclusion in Japanese high school students. This study used a sample of students from randomly selected urban and rural high schools in Japan's Kyushu district. The DAI scores for urban and rural high school students, as well as the total DAI scores from these two areas were significantly higher than that of American high school students. These results showed that the number of students with an acceptable dental appearance among Japanese high school students is significantly lower than among American high school students. This indicates a higher need for orthodontic treatment among Japanese high school students. PMID- 8222607 TI - Access to dental care--experiences of men with HIV infection in the United Kingdom. AB - A piloted self-complete questionnaire enquiring about dental experiences was administered to a consecutive sample of men with asymptomatic HIV infection attending a dedicated clinic in London. Forty-seven men completed questionnaires. Despite a high level of dental attendance before diagnosis of HIV infection, 60% of the men had not visited a general dental practitioner since diagnosis. Nineteen men had visited a dentist since diagnosis; 15 had been refused or deterred by members of the dental team and 5 had attended without disclosing their HIV status. These data indicate that despite professional guidance to dentists, the perceptions of dentists held by people with HIV and the behaviours of dentists acts as barriers to care. Dedicated dental clinics act as an important safety-net for people with HIV infection. PMID- 8222608 TI - Study of the dental health of children in remission from acute lymphoblastic leukaemia in Northern Ireland. AB - The dental health of 54 children in remission from acute lymphoblastic leukaemia was compared with a similar number of matched healthy control subjects. Frequency of dental attendance was similar in both groups but most children in remission from leukaemia attended a hospital dentist, whereas children in the control group visited a dentist in general practice or in the public health service. Children in remission from leukaemia used fluoride supplements significantly more frequently than their controls. There was no difference in plaque or gingivitis scores between groups. Children in remission from leukaemia had a significantly lower number of decayed primary teeth (d) than their matched controls but there were no significant differences between study and control groups of m, f or dmft values. There was a significantly greater number of extracted teeth (M) and a significantly greater overall decay experience (DMFT) in the permanent dentition of children in remission from leukaemia compared to their controls. There were no significant differences between study and control groups for D or F values. The mean number of sealed teeth per patient was significantly higher in the study than in the control group. PMID- 8222609 TI - Factors connected with alveolar bone resorption among institutionalized elderly people. AB - In this socio-dental investigation among 175 institutionalized somatically disabled edentulous elderly people correlations are described between the test subjects' degree of alveolar bone resorption, their gender and age, their duration of edentulousness, and the number of complete dentures worn. All test subjects were interviewed by dental students and clinically examined by dentists. The mandibular alveolar bone resorption was significantly stronger in women than in men and stronger in people who had been edentulous longer. The longer the test subjects had been edentulous and the greater their degree of mandibular alveolar bone resorption was, the more mandibular dentures they had worn. This study among institutionalized elderly people provides for the most part the same results as a previously presented study among much younger people. Obviously, in these respects there are no differences between these groups. PMID- 8222610 TI - Dental caries experience among 12-year-old children in the Gaza Strip. PMID- 8222611 TI - Prevalence of tongue lesions in 1013 Jordanian dental outpatients. AB - Over a period of 10 months, 1013 consecutive dental outpatients attending the Screening Clinic in Faculty of Dentistry, Jordan University of Science and Technology, in the city of Irbid were investigated for the presence of tongue lesions. All subjects were seeking treatment for caries and periodontal diseases and none was complaining of oral mucosal disorder. The results showed that 240 subjects (23.7%) had one or more tongue lesions. Geographic tongue and fissured tongue were detected in 6.8% and 11.4% of the examined population respectively, and an association of the two conditions was observed in 2.9% of the subjects. Hairy tongue was diagnosed in 3.4% of the subjects with significantly higher prevalence in males (P < 0.0005). A strong correlation was found between hairy tongue and smoking. PMID- 8222612 TI - Prevalence of malocclusion in Senegal. PMID- 8222613 TI - Pediatric fiberoptic video bronchoscopy: the use of computer interfacing. AB - Conventional video-recordings of pediatric bronchoscopic procedures are routinely performed in many centers. The limitations of conventional video-recordings include an inability to concurrently compare serially recorded images, lack of color fidelity of the displayed image, difficulty in image retrieval of archived video, and the inability to subject the image to mathematical analysis. We describe a computer interface which addresses each of these limitations. PMID- 8222614 TI - Using an integrated software package for clinical data analysis on a microcomputer. AB - An integrated software package was used effectively for entering, organizing and analyzing clinical research data on a microcomputer. Both the database and the spreadsheet components of the package were used in the process. The database component enabled a form to be created for entering the data. The spreadsheet component was used in the organization and analysis of data. Macros were written within the spreadsheet environment for the statistical analysis of data. The purpose of this paper is to illustrate how an integrated software like Symphony could offer features beyond simply the use of a spreadsheet for the analysis of research data. Also highlighted are the other useful features of the integrated software that are not directly related to data analysis. PMID- 8222615 TI - CORDAT II: a new program for data acquisition and on-line calculation of hemodynamic and regional myocardial dimension parameters. AB - The present paper describes a new computer program for data acquisition and on line evaluation of hemodynamic parameters. The hardware setup is based on a standard i486 personal computer equipped with an analog to digital converter and an additional display controller. During data acquisition the multitasking program automatically detects the beginning and the end of a cardiac cycle. Immediately after the recognition of a cardiac cycle the program calculates and displays user-defined hemodynamic parameters and mean values over a given time or a given number of beats. There is a high correlation between manually determined hemodynamic parameters and the parameters determined automatically by CORDAT II. PMID- 8222616 TI - Identification of peak V in brainstem auditory evoked potentials with neural networks. AB - A feature extractor for determining the latency of peak V in brainstem auditory evoked potentials (BAEPs) is presented in this paper. A feature extractor that combines artificial neural networks with an algorithmic approach is presented. It consists of a series of small neural networks that have to make simple decisions. Each neural network decides what part of the input pattern contains the peak, and the algorithm passes that part of the pattern to the next neural network; in this way the size of the input patterns decreases during the process, and the last neural network determines the exact location of the peak. An optimal configuration of neural networks could determine the latencies of peak V in all synthetic evoked potentials correctly. With real evoked potentials, the networks yield results that comply with the opinion of a human expert in 80 +/- 6% of the cases. PMID- 8222617 TI - Use of running fractal dimension for the analysis of changing patterns in electroencephalograms. AB - Running fractal dimensions were measured on four channels of an electroencephalogram (EEG) recorded from a normal volunteer. The changes in the background activity due to eye closure were clearly differentiated by the fractal method. The compressed spectral array (CSA) and the running fractal dimensions of the EEG showed corresponding changes with respect to change in the background activity. The fractal method was also successful in detecting low amplitude spikes and the changes in the patterns in the EEG. The effects of different window lengths and shifts on the running fractal dimension have also been studied. The utility of fractal method for EEG data compression is highlighted. PMID- 8222618 TI - Two computer programs using logit transformation for the analysis of S-shaped curves. AB - Two BASIC computer programs using logit transformation for the analysis of S shaped curves are presented. The first program estimates the median effective dose (ED50) of drugs. A maximum likelihood method similar to Finney's algorithm of probit analysis is applied. The second fits S-shaped curves to empirical data which cannot be transformed easily to proportions. Logit transformation proves to be a simple and accurate alternative to probit transformation when computing memory is limited or calculation speed is critical. PMID- 8222619 TI - Clustering on the basis of longitudinal data. AB - A menu-drive PC program, ZDIST, for computing the distances between the estimated polynomial growth curves of subjects who have been followed longitudinally is described, illustrated, and made available to interested readers. These distances can be computed on the basis of the individual growth curves themselves and/or from estimates of individuals' growth velocity and acceleration curves. The resulting distance matrices can be saved in ASCII format and subsequently imported into any clustering program which accepts this type of input, e.g. SYSTAT. PMID- 8222620 TI - A hardware-independent data acquisition package incorporating a simulation driver and simulation description language. AB - Device drivers for commercial data acquisition boards (DASH-16, DT2801) and a device driver for a virtual data acquisition board have been developed and incorporated in a single package for a multiple station anesthesia research laboratory. The package provides a compiler for a simulation description language which is used to program the virtual board. The package, written in C for the IBM PC, was designed to be easily extended with additional drivers. Applications using the package can control all supported devices without the programmer having to learn the details of the hardware. PMID- 8222621 TI - Efficacy of skin barrier creams (III). The repetitive irritation test (RIT) in humans. AB - 2 barrier creams (BC) were evaluated against the anionic detergent sodium lauryl sulphate (SLS) using a new human test model. In the repetitive irritation test (RIT) on human skin, the irritant SLS is applied to the ventral forearm of healthy volunteers daily for 2 weeks. 1%, 5%, and 10% SLS is exposed to the skin for 30 min, using a glass cup 2.5 cm in diameter. The BC is applied 30 min before the irritant. Cutaneous irritation is assessed on a score for erythema (0 to 5+), and quantified by various biophysical techniques: transepidermal water loss (TEWL) by evaporimetry, skin blood flow volume (BFV) by laser-Doppler velocimetry, and skin colour by colorimetry (La* value). 10 subjects were tested with SLS on one forearm without pretreatment (control) and with Taktosan Salbe as BC on the other forearm. A 2nd panel of 10 subjects was tested in the same way with SLS and Marly skin as BC. Taktosan Salbe was extremely effective in reducing the irritation by SLS: there were significant differences regarding all test parameters for 10% SLS in the 2nd week. The most differentiating parameter was TEWL, revealing statistical differences as early as the 1st week for 10% SLS and Taktosan Salbe, while the least differentiating sensitivity was found for La*. In contrast, there was no significant suppression of irritancy in any parameter with Marly skin, either in the 1st week or in the 2nd week with any concentration of SLS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222622 TI - Exposure, skin protection and occupational skin diseases in the glass-fibre reinforced plastics industry. AB - A total of 100 workers, 86 from the glass-fibre-reinforced plastics (GRP) industry, 11 from polystyrene production and 3 from polyester resin coating manufacture, were examined for occupational skin hazards and for evaluation of skin protection. The workers had been exposed to many chemicals. Those working in the GRP industry had also been exposed to glass fibre and to dust produced by finishing work. 94% used protective gloves. 22 workers, all employed in the GRP industry, had contracted occupational skin disorders. 6 had allergic and 12 irritant contact dermatitis. 4 workers had an accidental injury caused by a peroxide catalyst, fire, hot air and constant mechanical friction. Allergic dermatoses were due to natural rubber (latex) (4 cases) in protective gloves, phenol-formaldehyde resin (1 case) and cobalt naphthenate (1 case). Irritant hand dermatoses (5 cases) were caused by the combined hazardous effect of unsaturated polyester or vinyl ester resins, organic solvents, glass fibre and dust from finishing work on the skin. Other cases of irritant dermatoses (7 cases) were due to the dust, promoted by mechanical friction of clothes. Skin disorders in the GRP industry were common (26%) but the symptoms were mild and only 3 patients had been on sick leave because of occupational skin disease. PMID- 8222623 TI - Influence of irritants on lymph node cell proliferation and the detection of contact sensitivity to metal salts in the murine local lymph node assay. AB - Dimethyl sulfoxide (DMSO) and sodium lauryl sulfate (SLS) are known to cause irritation of the skin, and to enhance the penetration of chemicals into the epidermis. In the present study, the lymph node cell (LNC) proliferative response following exposure to irritants, such as SLS and DMSO, was examined in the murine local lymph node assay (LLNA). Exposure to DMSO or SLS aqueous solution induced a small increase in lymph node cell proliferation compared with aqueous solution alone. Exposure to SLS in DMSO caused a significant increase in LNC proliferation. Further, the effect of addition of the irritants in a vehicle on the detection of contact sensitivity to metal allergens was examined. Application of potassium dichromate and nickel sulfate in DMSO or SLS aqueous solution caused increases in LNC proliferation. Exposure to metal allergen with SLS in DMSO also induced a significant LNC proliferative response, but did not induce a significant increase in stimulation index (increase in 3H-thymidine incorporation relative to vehicle-treated control group). This was because of increased 3H thymidine incorporation following exposure to SLS-DMSO in the control group. These results suggest that irritants enhance the LNC proliferative responses to metal allergens. The use of SLS in aqueous solution is effective for the detection of sensitivity to water-soluble allergens, such as metal allergens, in the LLNA, as well as the use of DMSO as an application vehicle. PMID- 8222624 TI - Chromium content in human skin after in vitro application of ordinary cement and ferrous-sulphate-reduced cement. AB - The amount of chromium found in human skin after in vitro application of cement suspensions on full-thickness human skin in diffusion cells was investigated. Cement suspensions made from ordinary Portland cement or Portland cement with the chromate reduced with added ferrous sulphate were used. The cement suspensions were either applied on the skin surface under occlusion for 48 h or applied repeatedly every 24 h for 96 h. No statistically significant difference in chromium content of skin layers between skin exposed to ordinary Portland cement, skin exposed to cement with added ferrous sulphate and unexposed skin was observed, despite a more permeable skin barrier at the alkaline pH of the cement suspensions, i.e., pH 12.5. Increased chromium levels in epidermis and dermis were seen when ordinary Portland cement was applied as a suspension with added sodium sulphate (20%) on the skin surface for 96 h. The content of water-soluble chromium in ordinary Portland cement may vary due to the alkali sulphate content of the cement. PMID- 8222625 TI - Carvone: an overlooked contact allergen cross-reacting with sesquiterpene lactones? AB - The terpene l-carvone is one of the main constituents of spearmint oil. The sensitizing potential of l-carvone has been considered low, but it has occasionally caused contact allergy in users of spearmint toothpaste and chewing gum. l-Carvone is also an oxidation product of d-limonene that occurs in solvents used increasingly in industry. We included l-carvone 5% pet. in the standard patch test series. In the 1st year, 541 patients were tested and 15 (2.77%) had positive, and 12 doubtful positive (?+) reactions to l-carvone. The strongest reactions were observed in 9 patients with concomitant Compositae sensitivity. The key clinical features and other contact allergies of the patients are presented. When re-testing with l-carvone in the same or lower concentrations, only 2 out of 8 patients had positive reactions. Possible reasons for this discrepancy are discussed in terms of cross-reactions, concomitant sensitization, excited skin syndrome, irritancy and facilitated immunological response. PMID- 8222626 TI - Allergic contact dermatitis from para-tertiary-butylphenol-formaldehyde resin (PTBP-F-R) in Northern Ireland. AB - Over the past 5 years, 2270 patients with suspected allergic contact dermatitis were patch tested in our clinic to the European standard series of allergens. 7 patients (0.3%) showed a positive reaction (> or = +) to PTBP-F-R. In 3 (42.9%) of these patients, the allergen was traced to adhesives but no allergen source was determined in the remainder (57.1%). On the basis of these findings, it is questionable whether PTBP-F-R alone should routinely be included in the European standard series. PMID- 8222627 TI - Occupational allergic contact dermatitis caused by diethylenetriamine in carbonless copy paper. AB - Carbonless copy paper, or 'no carbon required' (NCR) paper, has often been implicated as the cause of skin, respiratory, or general symptoms, but allergy has been verified in only a few cases. A 43-year-old machinist whose work involved the manufacture of NCR paper developed occupational dermatitis on the hands. On patch testing, both the NCR paper and 1 of the chemicals used to produce the microcapsules of the NCR paper, namely diethylenetriamine (DETA), provoked an allergic reaction. Analysis of the paper showed that it contained enough DETA to induce allergic contact dermatitis. People who handle NCR paper and develop symptoms of contact dermatitis should be patch tested with DETA. PMID- 8222628 TI - Ethylmercuric chloride: the responsible agent in thimerosal hypersensitivity. AB - The causative agent of thimerosal allergy (sodium ethylmercury thiosalicylate) has not previously been thoroughly investigated. To evaluate whether the organic mercury component or the thiosalicylic acid molecule induces thimerosal sensitization, 23 patients positive to thimerosal were patch tested with ethylmercuric chloride, thiosalicylic acid and 8 different derivatives of mercury. To date, ethylmercuric chloride has not been tested in thimerosal allergy. 19/23 patients (82%) showed positive patch test reactions to ethylmercuric chloride. 4/23 patients negative to ethylmercuric chloride reacted positively to thimerosal 0.1% but not to thimerosal 0.05%. 8/23 patients (35%) also reacted to other mercurials. 20 controls negative to thimerosal showed negative patch test reactions to ethylmercuric chloride. Neither patients nor controls reacted to thiosalicylic acid. These results indicate that testing with thimerosal 0.1% leads to false-positive reactions and that the ethyl mercury component is the responsible agent in thimerosal allergy. PMID- 8222629 TI - Topical provocation of fixed drug eruption due to sulphamethoxazole. PMID- 8222630 TI - Contact allergy to penicillamine in eyedrops. PMID- 8222631 TI - Echographic evaluation of subclinical allergic patch test reactions. PMID- 8222632 TI - Hypertrophic amalgam dermatitis of the tongue simulating carcinoma. PMID- 8222633 TI - Contact urticaria from pentamidine isethionate. PMID- 8222634 TI - Dyshidrosiform pemphigoid induced by nickel in the diet. PMID- 8222635 TI - Subacute contact dermatitis from fepradinol. PMID- 8222636 TI - Contact stomatitis from tixocortol pivalate. PMID- 8222637 TI - Contact dermatitis from ketoconazole. PMID- 8222638 TI - Sensitivity to clioquinol and chlorquinaldol in the quinoline mix. PMID- 8222639 TI - Phototoxic contact dermatitis with toxic hepatitis due to the percutaneous absorption of paraquat. PMID- 8222640 TI - Occupational airborne contact dermatitis from Machaerium scleroxylon (Santos rosewood). PMID- 8222641 TI - Occupational airborne contact dermatitis due to benzoyl peroxide. PMID- 8222642 TI - Delayed hypersensitivity to stepronin: a case report. PMID- 8222643 TI - Contact sensitivity to piroxicam. PMID- 8222644 TI - Extracellular matrix proteins in bile and serum of patients with gallstone disease. AB - The relationship of basement membrane to interstitial collagen-related protein metabolism was investigated in a sample of 100 consecutive patients undergoing surgery for gallstone disease. The measurements were performed on both sera and bile specimens using specific radioimmunoassays for type IV collagen, laminin, endogenous intact human type III collagen aminopropeptide and its degradation product, Col. 1. While basement membrane related proteins, type IV collagen and laminin were dominant in the bile, type III collagen related proteins were lower than in the corresponding serum samples. Both the intact type III procollagen peptide and its Col. 1 fragment were, however, found in the bile. The highest bile laminin concentrations were observed in patients with gallbladder fibrosis, whereas type III aminopropeptides were elevated not only in fibrosis and cancer but, most markedly, in acute inflammation of the gallbladder. Bile type IV collagen concentration was also found to vary according to the cholesterol content of gallstones. The results point to differences in the metabolism of various extracellular matrix proteins during the development of gallstone disease. The association between such proteins, the histological alterations in the gallbladder wall and the cholesterol content of gallstones may have implications for the pathogenesis of gallstone disease. PMID- 8222645 TI - Inhibitory effects of hyaluronan on neutrophil-mediated cartilage degradation. AB - The effects of hyaluronan on neutrophil-mediated cartilage degradation were studied in an in vitro model. Rat peritoneal neutrophils were incubated for up to 18 h with the neutralized cartilage of bovine nasal septa using N-formyl-L methionyl-L-leucyl-L-phenylalanine combined with cytochalasin B, or opsonized zymosan, as a stimulation agent of neutrophils. Hyaluronan inhibited the neutrophil-mediated cartilage degradation by reducing the release of sulfated glycosaminoglycans from the cartilage. Inhibitory effects were dependent on concentration and molecular weight of hyaluronan. These results suggest that hyaluronan with a high molecular weight plays an important role in protecting the articular cartilage in inflamed joints from neutrophil injury. PMID- 8222646 TI - Collagen of accessory structures of organ of Corti. AB - It was previously demonstrated that about 40% of the protein of the tectorial membrane of the guinea pig consists of collagen type II, with lesser amounts of type IX and XI. In this paper we extend these studies on the tectorial membrane to the basilar membrane and to other accessory structures, the spiral ligament and spiral limbus. Earlier immunohistochemical data indicated that no collagen type II is present in the basilar membrane of the newborn guinea pig, but that it is present in the area of the basilar membrane in the embryo. However, by means of stringent extraction procedures we have determined biochemically that collagen type II and lesser amounts of type XI are present in the basilar membrane of the adult guinea pig, at similar levels (on the basis of total protein) to the tectorial membrane. Levels of collagen type II are much lower in the spiral ligament and spiral limbus. The presented studies demonstrate that classical techniques of collagen chemistry can be applied at the microscale on minute tissue elements. The significance of the presence of collagen in the tectorial membrane and basilar membrane is discussed in the light of known mechanical properties of these structures. PMID- 8222647 TI - Occurrence and localization of transforming growth factor-beta (TGF-beta 1, beta 2) during rabbit skin development. AB - The expression of (TGF beta-1) and (TGF beta-2) in adult, fetal, and neonatal rabbit skin has been analyzed by immunohistochemistry. Samples were collected from the backs of fetal rabbits (23 and 30 days of gestation), newborn rabbits (1 day post-partum) and adult pregnant and nonpregnant rabbits. Forty samples were studied. To identify TGF-beta two antibodies were used: (RAB4) and (CL-B1/29). TGF-beta 1 and TGF-beta 2 displayed distinct distribution and intensity of staining at different stages of development. TGF-beta 1 was present in fetal skin (23 days of pregnancy) expressing abundant staining in cells within epidermis, dermis and hypodermis. Fetal skin (30 days of pregnancy) exhibited weak staining for TGF-beta 1; only isolated dermal cells in close proximity to hair follicles and sweat glands expressed intracellular staining. Newborn rabbit skin showed low expression for TGF-beta 1, around hair follicles and sweat glands. In contrast adult pregnant and non-pregnant rabbit skin exhibited abundant staining for TGF beta 1 in epidermis, papillary dermis, around hair follicles, sweat glands, and blood vessels, no difference was observed among pregnant and non-pregnant rabbits. TGF-beta 2 was detected in all specimens and the distribution and intensity of staining were similar to those found for TGF-beta 1 in all groups. Our findings demonstrate that the expression of TGF-beta 1 and TGF-beta 2 is intense during early stages of rabbit skin development when morphogenesis is taking place and skin appendages are being formed; and less intense when these structures are formed and skin growth predominates. These results suggest an important role for TGF-beta 1 and TGF-beta 2 in rabbit skin development, probably regulating morphogenesis and growth. PMID- 8222648 TI - Relative efficiency of incorporation of newly synthesized elastin and collagen into aorta, pulmonary artery and pulmonary vein of growing pigs. AB - It is generally accepted that, once laid down in the extracellular matrix, elastin turns over only very slowly if at all. However, much of the evidence for minimal turnover of elastin comes from aortic tissue. In this study we have compared the relative synthesis rates of elastin and collagen with their relative accumulation rates in segments of aorta (AO), pulmonary artery (PA), and pulmonary vein (PV) of young, growing pigs. While rates of elastin synthesis are comparable in the AO and PA, the PA accumulates only 33% of the elastin deposited in the AO. Similarly, while the rate of elastin synthesis in the PV is 60% of that in the AO, accumulation of elastin is only 20% of that in the AO. Similar discrepancies between collagen synthesis and accumulation were seen among these three vessels. These data suggest that, at least in growing animals, the efficiency of permanent incorporation of elastin and collagen into the extracellular matrix may be an important factor in determining the final contents of these connective tissue proteins in vascular tissues. PMID- 8222649 TI - Direct extraction of gelatinases from rat bone. AB - Many studies have shown that gelatinases are secreted into the medium of cultures of various cell and tissue types, including bone cells. It is not clear, however, to what extent the culture process is responsible for inducing the expression of these proteases. In the present study, gelatinolytic enzymes were extracted directly from bone and other tissues and identified as bands of activity on SDS PAGE enzymograms using gelatin as the substrate. Two forms of gelatinase (72-kDa and 92-kDa) were present in extracts of normal young rat bone. Yields were markedly higher from compact bone than from other tissues (blood, marrow, tendon, cancellous bone, articular cartilage, and skin). More 92-kDa than 72-kDa gelatinase was extracted from bone. The proteolytic specificity of the 92-kDa gelatinase isolated from the bone extract was shown to be similar to that reported for the enzyme isolated from tissue culture media. Native type I collagen was not cleaved but heat denatured type I collagen (gelatin) and native type IV, type V, type IX and type XI collagens were degraded. The proteolytic activity was inhibited by EDTA. The results indicate that more gelatinases can be extracted from bone tissue than from other tissues using mild extraction conditions. The cellular origin and function of these enzymes in bone remain to be defined. PMID- 8222650 TI - Hemostatic and metabolic effects of lowering the ethinyl-estradiol dose from 30 mcg to 20 mcg in oral contraceptives containing desogestrel. AB - The metabolic and hemostatic effects of two oral contraceptives containing 150 mcg desogestrel and 20 mcg ethinyl-estradiol (EE) (MERCILON) or 30 mcg EE (MARVELON) were compared in order to examine the effect of reducing the EE dose in contraceptive pills. Forty-nine women participated in this randomized study during 6 cycles. In both groups, there was a significant increase in triglycerides, HDL-cholesterol and apoprotein A1; the same increase was observed for SBP and CBG. Slight and transient variations of fasting blood glucose levels were seen in the 30 mcg EE group and in the two groups for fasting insulin levels. The increase in renin substrate was significantly higher with the 30 mcg EE than with the 20 mcg EE pill. In both groups, plasminogen increased significantly, but antithrombin III, total and free protein S and fibrinogen decreased significantly only in women taking the 30 mcg EE pill, whereas there was no significant change in the 20 mcg EE group. Reducing the dose of EE in oral contraceptives from 30 mcg to 20 mcg minimizes their impact on renin substrate and hemostatic parameters. PMID- 8222651 TI - Long-interval treatment regimen with a desogestrel-containing oral contraceptive. AB - A randomized multicenter study was performed in order to investigate the acceptance of a low-dose OC (30 micrograms of ethinyloestradiol and 150 micrograms of desogestrel), using a 9 weeks on and 1 week off schedule (prolonged regimen, n = 198), compared to a traditional 3 weeks on, 1 week off schedule (standard regimen, n = 96). Haemoglobin and blood pressure remained the same in both groups during the study. No significant differences were found in body weight changes between the two groups. There was significantly more breakthrough bleeding and spotting in the group with prolonged regimen than in the group with standard regimen, but both breakthrough bleeding and spotting decreased during the trial. Irregular bleeding was significantly less in women who were already using OC, compared to "new starters." No serious side effects occurred. Significantly more women stopped the trial because of bleeding problems in the group with prolonged regimen, while there were significantly more women who stopped the trial because of headache in the group with standard regimen. After completing 12 months, or after premature withdrawal from the study, each women completed a questionnaire. Sixty-three per cent of the women preferred the studied alternative and twenty-six per cent preferred the traditional OC. PMID- 8222652 TI - Effects of a low-estrogen, desogestrel-containing oral contraceptive on lipid and carbohydrate metabolism. AB - Fasting serum lipids, lipoproteins and apolipoproteins, and fasting plasma glucose, insulin and C-peptide were measured in 107 non-users and 83 users of an oral contraceptive containing the progestin desogestrel, combined with 20 micrograms ethinyl estradiol. Plasma glucose, insulin and C-peptide concentrations during an oral glucose tolerance test (OGTT) were measured in a subgroup of 69 non-users and 39 users. Compared with non-users, users had higher concentrations of total, high density lipoprotein (HDL), HDL subfraction 3 and very low density lipoprotein (VLDL) cholesterol, total triglycerides, VLDL triglycerides, apolipoproteins AI and AII and fasting plasma insulin. There were no differences in HDL subfraction 2, low density lipoprotein cholesterol and apolipoprotein B. OGTT glucose was 60% higher in the users and OGTT insulin response 19% higher. The OGTT C-peptide response did not differ. The effects of 20 micrograms ethinyl estradiol combined with 150 micrograms desogestrel on lipid, lipoprotein, glucose and insulin metabolism are similar to those described previously with a 30 micrograms ethinyl estradiol combination containing the same dose of desogestrel. The relatively favourable metabolic profile associated with the higher estrogen dose desogestrel combination is maintained at the lower dose. PMID- 8222653 TI - One-year experience with subdermal contraceptive implants in the United States. AB - Women who received Norplant contraceptive implants from any of fifteen clinical settings in southeast Texas, U.S.A., were followed for one year to determine their reactions to the method. Of 1,385 who enrolled to receive Norplant implants, 1,253 had implants inserted. Side effects were reported by 78% of those receiving implants and 70% described changes in bleeding patterns. Spotting or irregular bleeding, weight gain and headaches were the conditions reported most frequently. Nine pregnancies were reported during the study period. Six of these, however, existed before the implants were inserted. At the one year anniversary, 143 of women receiving implants had had them removed. Those who discontinued method use were less satisfied, reported more side effects and were more likely to have planned to have another child, thus using the method for spacing, or to have had a change in their marital status while they were using the contraceptive. Providers should counsel patients to focus attention on plans for the future in selecting their contraceptive method. In addition, we recommend, as does the product's distributor, that providers confirm that patients are not pregnant prior to inserting implants. PMID- 8222654 TI - The presence of mast cells in the human endometrium pre- and post-insertion of intrauterine devices. AB - A study of the density of mast cells in the human endometrium was carried out on biopsy specimens prepared for light microscopy in 87 cases with three types of IUDs (stainless steel ring SS-type, copper Cu-T 220 and LNG-IUD levonorgestrel releasing device) pre- and post-insertion. The results revealed that, in general, there was an increase of mast cells after 3 to 24 months' use of IUD, independent of type (p < 0.01 when compared with the pre-insertion value). The increase in number of mast cells was most prominent in women having used the Cu-T220 IUD for 24 months (p < 0.05 when compared with the SS-IUD and p < 0.01 when compared with the LNG-IUD). The difference between the SS-IUD and the LNG-IUD as to the number of mast cells was not significant. No significant difference was found between the "bleeders" and "non-bleeders" in any of the three types of IUDs. It is noteworthy that patients using the Cu-T220-IUD had the highest percentage of patients with abnormal bleeding (> 50%) and that this group also had the highest density of mast cells per mm2 after 24 months' use. Although no significant difference with regard to the density of mast cells could be found between the "bleeders" and the "non-bleeders" in the present study, the number of "bleeders" of the three study groups is usually small and sometimes this number does not allow an adequate statistical analysis for comparison. Further investigations, involving larger number of IUD users, may yield a better understanding with regard to the potential relationship between the number of bleeding episodes and the density of mast cells in the human endometrium. PMID- 8222655 TI - Vasal sterilization in China. AB - Vasal sterilization has become the most popular method of contraception in many countries since the 1960's. In China, an estimated 30 million men have undergone voluntary sterilization. There have been two major developments in vasectomy technique, the no-scalpel method and the percutaneous chemical vas occlusion. These methods have significantly increased the acceptability of male sterilization in some cultures since no incisions in the skin are required. The effective rate is over 98% for both methods. The clients experience less pain, fewer complications and more rapid recovery. Epidemiological studies of large numbers of volunteers receiving no-scalpel or chemical vas occlusion procedures have resulted in a clearer picture of their safety, effectiveness, simplicity and economy. Meanwhile, some promising reversible vasal sterilization techniques have also been studied. PMID- 8222656 TI - Further acceptability evaluation of RU486 and ONO 802 as abortifacient agents in a Chinese population. AB - Of 144 consecutive women who requested early induced abortion, 99 (68.7%) and 45 (31.3%) women chose RU486 combined with ONO 802 (medical method) and suction evacuation (surgical method), respectively. Logistic regression analysis of covariates showed that age and marital status were significantly correlated with the acceptability and hence the choice of the medical method. There were also more working women in this medical group. Previous experience of induced abortion had no influence on the current choice of the abortion method. This group of women appeared to have a tendency of treating their disease with medication rather than with surgery if the condition would allow. They expressed fear about surgery. The long induction-abortion interval of three days will have to be tolerated, but the duration of bleeding should be minimised in order to improve the acceptability of the drug. RU486 is an alternative abortion method which should be made widely available. PMID- 8222657 TI - Fucoidin binding activity and its localization on human spermatozoa. AB - We previously reported that fucoidin (a polymer of predominantly sulfated L fucose) significantly inhibits: (1) tight binding of human sperm to human zona pellucida in vitro and (2) stimulation of the acrosome reaction by acid solubilized human zona pellucida. Here, we determined fucoidin binding activity on human spermatozoa and its localization on both live and permeabilized human sperm populations. A typical binding curve was demonstrated with biotinylated fucoidin. In competitive inhibition assays with unlabelled fucoidin or human sperm membrane extracts, IC50's were 4.0 micrograms/ml and 31.4 micrograms/ml, respectively. Fucoidin binding was localized over the acrosomal region of methanol-fixed human sperm and this pattern of binding significantly decreased from 92 +/- 3% to 74 +/- 6% with calcium ionophore A23187 treatment (p < 0.01). Binding of fucoidin-coated beads to live (non-permeabilized) human sperm was less than 1%. Addition of the detergent, Triton-X, to permeabilize sperm membranes resulted in a significant increase in binding (p = 0.001). These results provide evidence for the presence of a fucoidin binding compound in human spermatozoa that is localized to the membranes of the acrosomal region and can be extracted by a mild detergent extraction. Absence of binding by fucoidin to intact but not permeabilized spermatozoa suggests that the heteropolysaccharide binds to a receptor within the acrosomal matrix. However, further investigation is warranted to determine whether a fucoidin binding site is present both at the sperm's surface for the initial contact with the zona pellucida, and also for secondary binding after exposure of the acrosomal membranes. PMID- 8222658 TI - Comparison of two triphasic oral contraceptives containing either gestodene or norethindrone: a randomized, controlled trial. AB - Two triphasic oral contraceptives containing either gestodene or norethindrone as the progestogenic compound combined with ethinyl estradiol were compared in a randomized clinical trial to assess their contraceptive reliability, clinical tolerance and cycle control. Both preparations were effective in preventing pregnancy. The gestodene preparation proved significantly superior regarding cycle control and general tolerance. PMID- 8222659 TI - Pharmacokinetics of gestodene and ethinylestradiol in 14 women during three months of treatment with a new tri-step combination oral contraceptive: serum protein binding of gestodene and influence of treatment on free and total testosterone levels in the serum. AB - The pharmacokinetics of gestodene (GEST) and ethinylestradiol (EE2) were determined in 14 healthy women (age 18 to 32 years) during a treatment period of three months with a new tri-step combination oral contraceptive (Milvane). Prior to this treatment period, the same women received a single administration of a coated tablet containing 0.1 mg GEST together with 0.03 mg EE2. There was a wash out phase of one week between both treatments. Following single dose administration, a mean terminal half-life of 18 h was observed for GEST. The total clearance was 0.9 ml x min-1 x kg-1 and the volume of distribution was 84 l. During a treatment cycle, GEST levels in the serum accumulated by a factor of 8 as compared to single dose administration. Steady-state drug levels were reached during the second half of each cycle. As compared to single dose administration, the following changes were observed for GEST at the end of treatment cycles one and three: prolonged terminal half-life (20 to 22 h), reduced total (0.16 ml x min-1 x kg-1) and free clearance (ca. 27 ml x min-1 x kg 1), reduced volume of distribution (ca. 18 l). A concomitant EE2-induced increase in the SHBG concentrations by a factor of three as compared to pretreatment values was observed during a treatment cycle and appeared to be mainly responsible for the changes in the pharmacokinetics of GEST. Marked changes were also seen for the serum protein binding of GEST. After single dose administration, the free fraction of GEST was 1.3% and the fractions bound to SHBG and albumin were 69.4% and 29.3%, respectively. At the end of cycle one, the free fraction was only 0.6% and the fractions bound to SHBG and albumin were 81.4% and 18.0%, respectively. There was no difference in corresponding pharmacokinetic parameters and in the serum protein binding of GEST at the end of cycles one and three. On the last day of treatment cycles one and three, the AUC(0-4h) values of EE2 were 299.2 and 278.1 pg x ml-1 x h, respectively, which corresponds to an about 30% increase as compared to single dose administration, where an AUC(0-4h) value of 216.1 pg x ml-1 x h was found. Total and free testosterone concentrations decreased during treatment cycles one and three by about 36% and 60%, respectively, compared with the corresponding values measured prior to treatment. The fraction of unbound testosterone thus decreased from 0.5% to 0.3% during treatment. PMID- 8222660 TI - Safety and efficacy of a triphasic oral contraceptive containing desogestrel: results of three multicenter trials. AB - This report combines the data obtained in three multicenter efficacy studies with a new triphasic oral contraceptive combination containing desogestrel (DSG) and ethinyl estradiol (EE). The studies were conducted at 40 investigational sites in the United States (38 centers) and Canada (2 centers) in support of the FDA approval of the product. In total, 1,095 subjects were exposed to the study medication for 11,231 cycles, corresponding to approximately 864 woman-years of use. Of these subjects, 414 completed at least 13 cycles of treatment. Contraceptive efficacy was high; six pregnancies occurred during the in-treatment period (two due to method failure and four due to user failure), corresponding to Pearl Indices of 0.23 and 0.46, respectively. The incidence of irregular bleeding was low, and the acceptability was excellent as evidenced by the low incidence of drug-related drop-outs. No drug-related serious adverse experiences were reported, and the incidence of other drug-related adverse experiences was generally low and decreased with continued use. No effects were seen on blood pressure, body mass index, laboratory parameters, cervical cytology and breast nodularity. These studies demonstrate that triphasic DSG/EE is an effective and acceptable triphasic oral contraceptive preparation with excellent cycle control and no significant physiologic effects. PMID- 8222661 TI - Methotrexate and misoprostol for early abortion. AB - Methotrexate is cytotoxic to trophoblast and, in low doses, has minimal side effects. It is used to treat both gestational trophoblastic neoplasia and ectopic pregnancy. The cytotoxic effects of methotrexate on intrauterine trophoblast should be equivalent. To test this hypothesis, ten pregnant women, < 8 weeks' gestation were treated with methotrexate 50 mg/m2 intramuscularly followed 3 days later by misoprostol, a prostaglandin E1 analogue. The first 4 patients received misoprostol 600 micrograms orally; none aborted soon after the misoprostol. Two patients aborted 25 and 26 days after the methotrexate injection and two elected a suction abortion after 14 days (one by choice and one because the pregnancy was still viable). The last 6 patients received misoprostol 800 micrograms vaginally and aborted within 3-8 hours. One patient had an incomplete abortion requiring a suction curettage 34 days after the misoprostol. Vaginal bleeding for these 6 patients lasted an average of 29 +/- 11 days (range, 12-42 days). No methotrexate side effects were observed. Vaginal misoprostol (800 micrograms) was significantly more effective (p = 0.005) than oral misoprostol (600 micrograms) in effecting abortion after intramuscular methotrexate. PMID- 8222662 TI - Immediate postplacental insertion and fixation of the CuFix postpartum implant system. AB - Since promising results were obtained with the CuFix interval implant system in international clinical trials conducted worldwide, a modification of the device adapting it for immediate postplacental insertion and fixation (IPPIF) seemed a logical approach to solve the expulsion problem of IUDs inserted at that particular time. Seventy-three insertions were performed at three university centers, during 1990 and 1991, both by skilled and unskilled investigators. The results of this multicenter study show the excellent retainability of the modified IUD supporting the validity of the anchoring principle. The insertion procedure is easy, requiring minimal training, and the insertion technique appeared to be safe. PMID- 8222663 TI - A preliminary report of Norplant implant insertions in a large urban family planning program. AB - During the 21-month period between August 1, 1991, and April 30, 1993, 2,358 women received Norplant system insertions in either Parkland Memorial Hospital or the Dallas Maternal Health and Family Planning clinics. Forty-three percent of these women were teenagers with 14% 16 years of age or less. Overall, 431 patients received Norplant implants insertions postpartum prior to discharge from the hospital. To date, 138 Norplant systems have been removed, with the proportion of contraceptive implant removals among teenagers being essentially the same as that in more mature women. Of the reasons given by women discontinuing the Norplant system, an unanticipated high incidence of pain in the arm containing the implants, hair loss, and mood changes were noted. We have found the Norplant system to be a highly effective and highly acceptable contraceptive method for a large number of indigent women. PMID- 8222664 TI - Phase I clinical trial of an injectable contraceptive for the male. AB - Earlier studies on the rat and the monkey had demonstrated that an injection of styrene maleic anhydride (SMA) in a solvent vehicle of dimethyl sulphoxide (DMSO) into the lumen of the vas deferens is toxicologically safe and has contraceptive action. Phase I clinical trial was therefore undertaken on 38 male volunteers giving varying doses of SMA, ranging between 5 mg and 140 mg, into each vas deferens. A dose of 70 mg is the predicted therapeutic dose based on animal data. That the compound is within the vas deferens lumen during the period of the safety assessment is inferred from the effect on the spermatozoa count in ejaculates which reach azoospermic levels in the higher dose ranges. The treatment is well tolerated with only minimal side effects in a few cases and no long-term adverse effects. PMID- 8222665 TI - Absence of an effect of high vitamin C dosage on the systemic availability of ethinyl estradiol in women using a combination oral contraceptive. AB - Previous studies in small numbers of women have suggested that the administration of gram quantities of ascorbic acid interferes with the conversion of ethinyl estradiol (EE2) to its sulfates, leading to higher blood levels of EE2. The possibility of such potentiation has been investigated in 37 women using a combination monophasic oral contraceptive (30 micrograms EE2 and 150 micrograms levonorgestrel) for two consecutive cycles. Concomitant daily administration of 1 g ascorbic acid taken 1/2 hour before OC intake, was randomly assigned to the first or second cycle of OC use. On the first and 15th day of OC intake, blood samples were drawn 11 times over a 12-hour interval and Cmax and AUC(0-12 h) calculated. On pill days 10 and 21, only 6-hour post-intake samples were obtained. Samples were analyzed for levels of ascorbic acid, free and sulfated ethinyl estradiol (and a number of other parameters). Cmax and AUC values for EE2 and EE2-sulfate in cycles with and without ascorbic acid were evaluated statistically by the Grizzle model for days 1 and 15 and the ratios of day 15/day 1 for each of the substances. No effect of ascorbic acid was observed (alpha = 0.05, 1-beta = 0.9). Only on day 15 was there a significantly lower AUC for EE2 sulfate in the presence of ascorbic acid intake. Thus, the competition between ascorbic acid and EE2 for sulfation does not lead to an increased systemic availability of EE2 and is, therefore, unlikely to be of any clinical importance. Ascorbic acid can, therefore, be removed from the list of drugs interfering with the pharmacokinetics of ethinyl estradiol. PMID- 8222666 TI - Suspended Judgment. From "tame" to "forbidden" death. PMID- 8222667 TI - Statistical considerations in monitoring the Systolic Hypertension in the Elderly Program (SHEP). AB - The Systolic Hypertension in the Elderly Program (SHEP), a randomized, double masked, placebo-controlled trial of 4736 persons, was designed to assess the efficacy of antihypertensive drug treatment to reduce the risk of fatal and nonfatal strokes among people age 60 and over with isolated systolic hypertension. The statistical method used in interim monitoring of results was conditional power (or stochastic curtailment). The findings did not become conclusive until near the completion of the trial, and therefore SHEP was continued to its scheduled closing date. The trial demonstrated a 36% reduction in the incidence of stroke in the active treatment group (P = .0003). In addition to evaluating overall efficacy of treatment, the monitoring process considered such other issues as nonstroke outcomes, lag time between first report of stroke and final confirmation of stroke diagnosis, consistency of results across subgroups, and completeness of follow-up. The purpose of this article is to review these factors with primary emphasis on the statistical aspects. PMID- 8222668 TI - Design and methods of a clinical trial for a rare condition: the Collaborative Ocular Melanoma Study. COMS Report No. 3. AB - Investigators who conduct clinical trials of treatments for uncommon conditions face special challenges regarding trial design and execution in addition to the challenges faced by all clinical trial investigators. The Collaborative Ocular Melanoma Study (COMS) currently consists of two multicenter, randomized controlled clinical trials designed to investigate the efficacy of radiotherapy compared to surgery in prolonging the survival of patients with choroidal melanoma, a rare intraocular cancer. Patients with unilateral choroidal melanoma classified as "medium" in size are randomized with equal probability to either enucleation (removal of the eye) or radiation delivered to the tumor by means of a radioactive "plaque" attached by sutures to the scleral surface of the eye over the base of the tumor. Patients with large tumors are randomized with equal probability to either enucleation or a 5 day course of external beam radiation therapy followed by enucleation. Time to death is the primary outcome; patients will be followed for at least 10 years or until death. Quality assurance mechanisms for evaluation, treatment, and follow-up have been implemented. This paper describes the design and methods of the trials, highlights the challenges associated with implementing and conducting the study, and summarizes the current status of the study. PMID- 8222669 TI - Outcome of the elderly after out-of-hospital cardiac arrest. PMID- 8222670 TI - Too much pharmacokinetics? PMID- 8222671 TI - Delivery dependent oxygen consumption: asking the wrong questions and not getting any answers. PMID- 8222672 TI - Hemodynamic monitoring--a new day dawning. PMID- 8222673 TI - Multiple organ failure in burn patients. PMID- 8222674 TI - Selective decontamination of the digestive tract: theory or therapy? PMID- 8222675 TI - Will the real culprit please stand up! PMID- 8222676 TI - Designer lipids as a nutraceutical. PMID- 8222677 TI - A double-blind, prospective, randomized trial of ketoconazole, a thromboxane synthetase inhibitor, in the prophylaxis of the adult respiratory distress syndrome. AB - OBJECTIVE: To determine if ketoconazole, a thromboxane A2 synthetase inhibitor, given within the first 24 hrs after diagnosis and arrival in the intensive care unit (ICU) would decrease the frequency of adult respiratory distress syndrome in the septic patient population. DESIGN: Prospective, randomized, double-blind, placebo-controlled study. SETTING: Twelve-bed, surgical ICU in a university affiliated hospital. PATIENTS: Fifty-four consecutive patients admitted to the surgical ICU with the diagnosis of sepsis composed the study sample. Sepsis was defined as including two or more of the following signs in a patient with a systolic blood pressure of < 80 mm Hg or a systemic vascular resistance of < 800 dyne.sec/cm5: a) temperature > or = 39 degrees C or < or = 35 degrees C; b) white blood cell count of > 12,000 leukocytes, or < or = 4000 leukocytes/microL, or > or = 20% immature cells; c) positive blood culture; d) known or strongly suspected source of infection from which a known pathogen was cultured. INTERVENTIONS: Patients were randomized to receive either ketoconazole (400 mg) or placebo in a double-blind fashion as early as possible and in < 24 hrs after surgical ICU admission or after the diagnosis of sepsis was established. MEASUREMENTS AND MAIN RESULTS: Adult respiratory distress syndrome (ARDS) was diagnosed if the following criteria were met: a) intrapulmonary shunt of > 20%, or a PaO2/FIO2 ratio of < 150 requiring ventilatory support for > 48 hrs; b) pulmonary artery occlusion pressure of < 18 mm Hg and no clinical signs of heart failure; and c) diffuse infiltrates on chest radiograph. Treatment resulted in significant (p = .002) reduction in the frequency of ARDS compared with the placebo group, 64% vs. 15% in the ketoconazole treated group. The mortality rate was also reduced from 39% in the placebo group to 15% in the ketoconazole group (p = .05). A statistically significant reduction in ventilator and ICU days was not achieved. CONCLUSIONS: Ketoconazole (400 mg through the gastrointestinal tract) given early in the septic course may prevent ARDS and decrease the mortality rate in high-risk, septic patients. PMID- 8222678 TI - Ketoconazole: maybe it isn't the magic potion, but.... PMID- 8222679 TI - Survival in the elderly after out-of-hospital cardiac arrest. AB - OBJECTIVES: To evaluate the survival prognosis for the elderly (> or = 70 yrs of age) after out-of-hospital cardiac arrest in a large urban center, and to identify any specific differences in survival factors relative to those adults < 70 yrs of age. DESIGN: The study was a prospective, inception cohort study. SETTING: An urban population of approximately 2,000,000, served by one centralized municipal emergency medical services system. PATIENTS: All 986 adult victims (367 elderly and 619 younger patients) of primary cardiac arrest attended by the emergency medical services system over a 12-month period. INTERVENTIONS: Not applicable. MEASUREMENTS AND MAIN RESULTS: All victims of out-of-hospital cardiac arrest occurring within a single, large, urban municipality were studied over a 12-month period. Each event was analyzed for age, sex, witnesses, bystander cardiopulmonary resuscitation, presenting electrocardiographic rhythm, paramedic response time, scene time, return of spontaneous circulation (pulses), and electrocardiographic rhythm on hospital arrival. Outcomes evaluated included inhospital admission (resuscitation) and successful discharge from the hospital (survival). Patients were followed until death or discharge from the hospital. Of 367 elderly cardiac arrest victims, 81 (22%) patients were successfully resuscitated and 24 (7%) patients survived. However, of 119 (32% of all elderly patients) patients who presented with ventricular fibrillation/tachycardia, 48 (40%) patients were resuscitated and 17 (14%) patients survived. These 17 patients with ventricular fibrillation/tachycardia accounted for 71% of all elderly survivors. During the same study period, there were 619 adult primary cardiac arrest victims < 70 yrs of age, 160 (26%) of whom were resuscitated and 73 (12%) of whom survived. Among the younger patients, 296 (48%) patients presented with ventricular fibrillation/tachycardia, of whom 110 (37%) were resuscitated and of whom 60 patients (20%) survived. Within the context of this study, survival rates for younger and older ventricular fibrillation/tachycardia patients were not significantly different. Also, among survivors, there were no other major differences in terms of established survival determinants. CONCLUSIONS: Survival chances for the elderly after out-of-hospital cardiac arrest are not bleak, and are reasonable if ventricular fibrillation/tachycardia is the presenting rhythm. Survival determinants are similar for younger and older adults. PMID- 8222680 TI - Steady-state pharmacokinetics of dopamine in adult patients. AB - OBJECTIVE: To study the pharmacokinetics of dopamine in hemodynamically stable adult patients. DESIGN: Prospective clinical study. SETTING: University hospital intensive care unit. PATIENTS: Fourteen patients (aged 43 to 73 yrs) recovering from esophageal surgery. INTERVENTION: Dopamine was infused and blood samples were collected. MEASUREMENTS AND MAIN RESULTS: Plasma dopamine concentrations were measured at steady state and subsequently at the end of the dopamine infusion using high-performance liquid chromatography. Clearances, volume of distribution, mean residence times, half-lives, and elimination and distribution rate constants were derived. The clearances were independent of the infusion rate at 1, 3, and 6 micrograms/kg/min, and ranged between 0.050 and 0.056 L/min/kg. A two-compartment open model was fitted to the postinfusion plasma concentration data obtained at 3 and 6 micrograms/kg/min. On average, the steady-state volume of distribution and the apparent terminal elimination half-life increased with the dose: 0.78 to 1.58 L/kg, respectively, and 22.1 to 37.9 mins, respectively, for the rates of 3 and 6 micrograms/kg/min. The rate constant associated with the uptake of dopamine into the peripheral compartment (K12) was on average four to five times higher than the rate constant associated with the reverse process (K21). CONCLUSIONS: The redistribution of dopamine into the central compartment could be the main factor involved in the apparent terminal elimination of dopamine from plasma. Due to the relative rates of distribution and elimination, the attainment of a steady-state plasma concentration of dopamine should only depend on the terminal half-life. These results, which remain to be validated in a greater number of patients, indicate that the attainment of 90% of the plateau (i.e., in 3.3 half-lives) would require 70 to 125 mins, depending on the infusion rate. PMID- 8222681 TI - Relationship between oxygen uptake and oxygen delivery in septic patients: effects of prostacyclin versus dobutamine. AB - OBJECTIVE: To compare the effects of prostacyclin (PGI2) and dobutamine on the relationship between oxygen delivery (DO2) and oxygen uptake (VO2) in stable septic patients. DESIGN: Prospective study using a crossover design with alternate order of medications. PATIENTS: Seventeen patients with documented sepsis and a stable hemodynamic status with normal blood lactate concentrations. Eleven patients were mechanically ventilated. Eight patients eventually died. INTERVENTIONS: DO2 and VO2 were calculated before and after a 20-min infusion of 5 ng/kg/min of PGI2 followed or preceded by 5 micrograms/kg/min of dobutamine. MEASUREMENTS AND MAIN RESULTS: Both medications increased cardiac output significantly. At the dose used, PGI2 infusion reduced mean arterial pressure from 90.8 +/- 16.8 to 81.5 +/- 17.3 mm Hg (p < .01) and PaO2 from 97 +/- 25 torr to 82 +/- 22 torr (from 12.9 +/- 3.3 to 10.9 +/- 2.9 kPa) (p < .01) and increased venous admixture from 17.5 +/- 0.6% to 23.8 +/- 8.2% (p < .01). Dobutamine had no significant influence on these variables. PGI2 increased DO2 by 19% (from 470 +/- 105 to 557 +/- 117 mL/min/m2, p < .01) while dobutamine increased DO2 by 27% (from 463 +/- 103 to 589 +/- 156 mL/min/m2, p < .01). PGI2 increased VO2 by 5% (from 148 +/- 38 to 155 +/- 36 mL/min/m2) while dobutamine increased VO2 by 10% (from 146 +/- 36 to 161 +/- 41 mL/min/m2, p < .01). Accordingly, there was an identical decrease in oxygen extraction with PGI2 (from 32.4 +/- 8.2% to 28.6 +/- 7.1%, p < .01) and dobutamine (from 32.4 +/- 8.3% to 28.5 +/- 7.8%, p < .01). The responses to these medications were similar in survivors and nonsurvivors. CONCLUSIONS: PGI2 and dobutamine at the doses used have similar effects on oxygen extraction in critically ill, septic patients, but dobutamine increases DO2 more consistently and is better tolerated than prostacyclin. PMID- 8222682 TI - Accurate assessment of right ventricular function in acute respiratory failure. AB - OBJECTIVE: Since right ventricular ejection fraction is highly dependent on afterload, right ventricular ejection fraction may not reflect right ventricular contractile function in acute respiratory failure. Despite a severe reduction in right ventricular ejection fraction, the right ventricle may be able to generate pressure output that is sufficient enough to maintain an adequate distribution of pulmonary perfusion. We tested this hypothesis by assessing the correlation between the right ventricular ejection fraction and the right ventricular end systolic pressure-volume relationship, and by assessing the correlations between right ventricular ejection fraction and the physiologic deadspace/tidal volume ratio and between the physiologic deadspace/tidal volume ratio and the right ventricular end-systolic pressure-volume relationship. DESIGN: Prospective study. SETTING: University hospital intensive care unit (ICU). PATIENTS: Twenty-one patients with acute respiratory failure. MEASUREMENTS AND MAIN RESULTS: The physiologic deadspace/tidal volume ratio was used as an index of the distribution of pulmonary perfusion. Right ventricular ejection fraction was measured by the thermodilution method. Right ventricular end-diastolic volume index was obtained from the stroke volume index divided by the right ventricular ejection fraction. Right ventricular end-systolic volume index was calculated as the difference between the right ventricular end-diastolic volume index and the stroke volume index. Pulmonary arterial dicrotic notch pressure was used as an estimate of right ventricular end-systolic pressure. Data were collected at baseline and after one or two alterations in preload to define the right ventricular end systolic pressure-volume relationship. There was no correlation between the right ventricular ejection fraction and the slope of the right ventricular end-systolic pressure-volume relationship line. No correlation was found between the right ventricular ejection fraction and the physiologic deadspace/tidal volume ratio. There was a hyperbolic curvilinear relationship between the physiologic deadspace/tidal volume ratio and the slope of the right ventricular end-systolic pressure-volume relationship line (r2 = .82, p < .0001). When the patients were divided into two groups based on the slope of the right ventricular end-systolic pressure-volume relationship line, the physiologic deadspace/tidal volume ratio was lower in the group with a high slope of the right ventricular end-systolic pressure-volume relationship line (p < .0001). There was no difference in other hemodynamic data between the two groups. CONCLUSIONS: These data suggest that in acute respiratory failure, the right ventricular ejection fraction does not reflect right ventricular performance. PMID- 8222683 TI - Multiple organ failure in patients with thermal injury. AB - OBJECTIVE: To assess the frequency and significance of multiple organ failure in patients with burn injuries. DESIGN: Retrospective review and prospective assessment of patients with acute burns. SETTING: University hospital burn center. PATIENTS AND METHODS: We reviewed 529 patients admitted for acute burn treatment whose lengths of stay exceeded 72 hrs. A new scoring system, the Thermal Injury Organ Failure Score, was used to assign scores from 0 (normal) to 6 (severe dysfunction) to each of 6 organ systems, which were then totaled to compile the overall score. This system was also used for prospective assessment of 83 adult burn patients, and compared with the Acute Physiology and Chronic Health Evaluation (APACHE) II scoring system during the first week of treatment. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: For 496 survivors, mean organ failure score was 3.28, compared with 23.1 in 33 nonsurvivors (p < .0001). All nonsurvivors but one had scores of > or = 15, indicating dysfunction of at least three organs. Scores and mortality rate increased with age and burn size. Pulmonary dysfunction was the most frequent form of organ failure seen, but correlated less with outcome than did cardiovascular or neurologic scores. Sepsis was present in 22 of 33 patients who died. In the prospective study, organ failure scores correlated with outcome more closely than did APACHE II scores. Weekly evaluation of these patients demonstrated progressive divergence in scores between survivors and nonsurvivors. CONCLUSIONS: Multiple organ failure was almost invariably present in burn patients who died > 72 hrs after injury. Burn victims, who have been excluded from reviews of multiple organ failure, appear to manifest organ failure in a manner similar to that of other surgical populations. The scoring system reported here may prove useful in evaluating organ failure in thermally injured patients. PMID- 8222684 TI - Can selective digestive decontamination avoid the endotoxemia and cytokine activation promoted by cardiopulmonary bypass? AB - OBJECTIVE: To evaluate the effect of selective digestive decontamination on endotoxemia and cytokine activation during the ischemic phase of cardiopulmonary bypass surgery. DESIGN: Prospective, open, randomized, controlled trial. SETTING: Two multidisciplinary intensive care units in tertiary care hospitals. PATIENTS: Eighty consecutive patients randomly allocated to two groups: selective digestive decontamination (group 1, n = 40) and controls (group 2, n = 40). INTERVENTIONS: Preoperative administration of oral antibiotics (polymyxin E, tobramycin, and amphotericin B) vs. untreated controls. MEASUREMENTS AND MAIN RESULTS: Assessment of decontamination by bacteriologic evaluation of rectal swabs (colony-forming units [cfu]/mL) were made in each group, along with circulating endotoxin, tumor necrosis factor and interleukin-6 (IL-6) determinations before surgery, during ischemic and reperfusion phases of bypass, and at 4 hrs and at 24 hrs after surgery. Group 1 patients showed that rectal bacteria decreased ten-fold after treatment for 24 hrs, thousand-fold after 48 hrs, and registered 0 cfu/mL after digestive decontamination was administered for > 72 hrs. Endotoxin and IL-6 assays showed significantly lower values in this latter group vs. those circulating concentrations of control patients. On the other hand, both endotoxin and IL-6 concentrations correlated positively with the duration of surgical ischemia. CONCLUSIONS: Selective digestive decontamination reduces the gut content of enterobacteria, with complete elimination after 3 days of treatment. This fact could explain the lower endotoxin and cytokine concentrations found in the blood samples of patients who had been fully decontaminated. Duration of aortic cross-clamping is an important factor in generating endotoxemia and in the activation of cytokines. PMID- 8222685 TI - Decreased mortality rate and length of hospital stay in surgical intensive care unit patients with successful selective decontamination of the gut. AB - OBJECTIVE: Current studies concerning selective decontamination of the digestive tract have failed to demonstrate a decrease in the length of hospital stay and mortality rate, despite the finding of a significantly lower number of infections. To evaluate this issue in more detail, the relationship between the mortality rate and length of stay with respect to colonization and infections was studied within a group of patients receiving selective decontamination. Special attention was given to the efficacy of decontamination within each patient. The main question addressed was whether an effect on mortality rate was present, and if so, why this effect was not apparent until now. DESIGN: Prospective observational cohort study. SETTING: Surgical intensive care unit (ICU) in a university hospital. PATIENTS: Ninety-seven patients primarily admitted into the surgical ICU who received selective decontamination. Transferred patients were excluded. The majority of the surgeries were elective, and all patients completed the follow-up. INTERVENTIONS: All patients received polymyxin E, amphotericin B, and norfloxacin four times a day in a 2% solution of Orabase orally and enterally as suspensions of 200, 500, and 50 mg, respectively. Assessment of the efficacy of selective decontamination was done by identification of Gram-negative microorganisms in surveillance cultures from the oropharynx and rectum. Predicted mortality rates for each patient were calculated with a logistic regression formula. MEASUREMENTS AND MAIN RESULTS: A possible benefit of selective decontamination of the digestive tract would be expressed by lower actual mortality rates compared to predicted mortality rates. Since we expected the efficacy of decontamination to have an influence on infection and mortality rates, we evaluated these rates in terms of successful or unsuccessful decontamination. Most patients (n = 72) were successfully decontaminated. Actual death rates in these patients were significantly lower than the expected rates (as calculated by the Acute Physiology and Chronic Health Evaluation [APACHE] II scoring system) (18% vs. 40%, p = .006), whereas no difference was found in those patients with failed decontamination (n = 25, death rate 44%). The patients with unsuccessful selective decontamination had significantly longer hospital (52 vs. 34 days) and ICU lengths of stays (23 vs. 9 days; p = .002) and higher mortality rates (44% vs. 18%, p = .020) when compared with those patients who were successfully decontaminated. CONCLUSIONS: These results indicate that selective decontamination is beneficial in terms of mortality rate and length of stay in surgical patients only when successful decontamination has been achieved. The subgroup of patients for whom decontamination is not successful might be responsible for the obscurity in mortality effects of selective decontamination in studies until now. It is expected that identification and subsequent elimination of possible risk factors that cause a failure of selective decontamination can result in lower morbidity and mortality rates in critically ill, surgical patients admitted to the ICU. PMID- 8222686 TI - Fatal course in severe meningococcemia: clinical predictors and effect of transfusion therapy. AB - OBJECTIVE: To investigate whether the administration of fresh-frozen plasma to patients with systemic meningococcal disease is associated with an increased mortality rate compared with the administration of plasma substitutes. DESIGN: Seventeen-year case-control study. SETTING: Intensive care units and departments of internal medicine and pediatrics of one university hospital and one local hospital. PATIENTS: A total of 336 patients with culture-proven meningococcemia or symptoms characteristic of meningococcemia who were admitted to two hospitals in northern Norway between 1974 and 1991. MEASUREMENTS AND MAIN RESULTS: High risk patients were selected on the basis of two different scoring systems (Niklasson's score and clinical score) and classified according to the type of intravenous fluid regimen (fresh-frozen plasma, blood, or colloids). For comparison between groups, analysis of variance and chi-square tests were used. Assessments of adjusted effects on mortality rate were done by multiple logistic regression. Administration of blood or plasma was significantly associated with a fatal course, both in the total patient population (p < .01) and in the high-risk group (p = .02), while using colloids alone was negatively associated with death, although not reaching statistical significance. A significantly lower mortality rate was found in one of the hospitals where colloids were used instead of plasma or blood in the last part of the period studied (p < .05). CONCLUSION: The results support our hypothesis that the use of fresh-frozen plasma may negatively influence outcome in systemic meningococcal disease. PMID- 8222687 TI - Intermediate syndrome in organophosphorus poisoning: a prospective study. AB - OBJECTIVES: To estimate the frequency of the intermediate syndrome in organophosphorus-poisoned patients, and examine its relationship to cholinesterase inhibition and electromyographic findings. Muscle biopsies were available in some patients. DESIGN: A 3-yr prospective study. SETTING: University teaching hospital intensive care unit. PATIENTS: Consecutive patients with acute organophosphorus poisoning (n = 19). MEASUREMENTS AND MAIN RESULTS: We determined the frequency of the intermediate syndrome in poisonings with various organophosphates, duration of (acetyl) cholinesterase inhibition and metabolite excretion, evolution of alterations on repetitive nerve stimulation, type and frequency of muscle lesions. A total of eight of 19 patients developed an intermediate syndrome. In some patients, short relapses of muscarinic symptoms superimposed on the intermediate syndrome. Agents such as methylparathion, fenthion, and dimethoate carry a high risk, but we also noted a prolonged intermediate syndrome in an ethyl-parathion-poisoned patient. Prolonged and severe cholinesterase inhibition occurred during the intermediate syndrome in all patients, and metabolite excretion was prolonged. As the intermediate syndrome evolved, repetitive nerve stimulation initially demonstrated decrement, then increment, and finally, normal responses. Necrotic fibers were noted in muscle biopsies, but these fibers were too sparse to explain severe muscle weakness and were similar in patients with and without the intermediate syndrome. No patients developed delayed neuropathy. CONCLUSIONS: The intermediate syndrome is not rare. Although it is more likely to occur with some organophosphates, it is not confined to a few distinct compounds. This syndrome coincides with prolonged cholinesterase inhibition, and is not due to muscle fiber necrosis. When viewed together, the clinical and electromyographic features are best explained by combined pre- and postsynaptic dysfunction of neuromuscular transmission. The intermediate syndrome is not related to an incipient delayed neuropathy. PMID- 8222688 TI - Derangements in peripheral glucose and oxygen utilization induced by catabolic hormones. AB - OBJECTIVE: To assess if the derangements in peripheral glucose, lactate, and oxygen utilization that are observed in severely injured patients are due to the hormonal response to injury or are related to the extent of the wound. DESIGN: Comparison study. INTERVENTIONS: The catabolic hormones epinephrine, cortisol, and glucagon were infused simultaneously into the femoral artery of six healthy volunteers, thus simulating the hormonal milieu associated with severe trauma in an uninjured leg. SETTING: Clinical research center at a university-affiliated hospital. PATIENTS: Young, adult males deemed healthy by screening medical history, physical examination, and blood chemistries. MEASUREMENTS AND MAIN RESULTS: Substrate net balance and indirect calorimetry measurements were performed before and then at the completion of 2 hrs of catabolic hormone infusion. Catabolic hormones elicited significant increases in leg glucose uptake and oxidation, and an increased net efflux from the leg of lactate and alanine. While leg oxygen delivery also increased, catabolic hormones failed to alter peripheral oxygen consumption. CONCLUSIONS: Catabolic hormones can elicit a similar peripheral metabolic response in an uninjured leg as that reported previously by Wilmore et al. in severely burned extremities. This finding suggests that the hormonal milieu associated with severe injury is influential in regulating peripheral glucose and oxygen utilization and that wound inflammation is not an essential component of this response. PMID- 8222689 TI - Ventilatory drive and carbon dioxide response in ventilatory failure due to myasthenia gravis and Guillain-Barre syndrome. AB - OBJECTIVE: To test the hypothesis that either decreased ventilatory drive or decreased CO2 responsiveness accounts for the hypoventilation observed in patients during acute ventilatory failure from myasthenia gravis or Guillain Barre syndrome. DESIGN: Prospective, consecutive case series evaluating trials of ventilatory muscle performance, ventilatory drive, and CO2 response in patients during recovery from ventilatory failure until they were weaned from mechanical ventilation. SETTING: Neurosciences critical care unit in a university hospital. PATIENTS: Seven intubated, mechanically ventilated patients with myasthenia gravis or Guillain-Barre syndrome. INTERVENTIONS: Patients repeatedly performed mechanically unsupported, spontaneous breathing trials to the limits of endurance. After spontaneous breathing trials, patients underwent CO2 rebreathing studies. MEASUREMENTS AND MAIN RESULTS: Seventy-three breathing trials were performed in three patients with Guillain-Barre syndrome and four patients with myasthenia gravis. Patients were unable to sustain spontaneous ventilation in 55 trials averaging 27 +/- 5 mins. In these trials, significant increases occurred in mean end-tidal CO2 (41 +/- 1 to 44 +/- 1 torr [5.6 +/- 0.1 to 6.0 +/- 0.1 kPa]) and respiratory rate (31 +/- 1 to 35 +/- 1 breaths/min, p < .01). Ventilatory drive (as measured by airway occlusion pressure for 100 msecs) increased significantly p < .01 from 3.7 +/- 0.3 to 4.9 +/- 0.3 cm H2O. The response of airway occlusion pressure to CO2 rebreathing after these trials was 0.33 +/- 0.07 cm H2O/sec/mm Hg, while the minute ventilation response to CO2 rebreathing was only 0.30 +/- 0.06 L/min/mm Hg. CONCLUSIONS: These results suggest that ventilatory drive increases during acute hypoventilation, and the ventilatory drive response to CO2 remains intact, even when the minute ventilation response to CO2 is poor. Therefore, a decrease in ventilatory drive or CO2 response is unlikely to account for hypoventilation during ventilatory failure in patients with myasthenia gravis or Guillain-Barre syndrome. PMID- 8222690 TI - Endotracheal intubation and mechanical ventilation in severe asthma. AB - OBJECTIVE: To determine the occurrence rate of complications and mortality in patients with severe asthma requiring endotracheal intubation and mechanical ventilation. DESIGN: Retrospective review of medical records from September 1982 to July 1988. SETTING: Urban, teaching hospital serving primarily indigent patients. PATIENTS: Fifty-seven adult patients with asthma requiring tracheal intubation and mechanical ventilation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Fifty-seven patients requiring tracheal intubation and mechanical ventilation during 69 hospital admissions were identified. Medication noncompliance and upper respiratory tract infections were recorded as the most frequent precipitating events for exacerbation of asthma. Forty-nine intubations were initiated because of a clinical diagnosis of respiratory distress, but multiple indications were present in 42 admissions. One or more complications occurred in 31 episodes of endotracheal intubation and mechanical ventilation (45%). Death occurred in four (6%) of 69 admissions. Three of the four deaths occurred in patients who had a cardiorespiratory arrest before hospital admission. CONCLUSIONS: While complications occurred in 45% of patients with severe asthma requiring intubation and mechanical ventilation, the mortality rate was low. We conclude that intubation and mechanical ventilation in patients with life-threatening asthma are safe and beneficial interventions. PMID- 8222691 TI - Myocardial contractility in a canine model of the brain-dead organ donor. AB - OBJECTIVE: To test the hypothesis that the induction and maintenance of brain death leads to a decrease in myocardial contractility. DESIGN: Prospective, randomized, controlled trial. SETTING: Laboratory at a large, university affiliated medical center. SUBJECTS: Sixteen adult, male, mongrel dogs weighing 15 to 25 kg. INTERVENTIONS: Myocardial contractile performance was evaluated after the induction of either brain death (n = 8), sham brain death (n = 4), or an operative procedure serving as a time control (n = 4). Brain death was induced by increasing and maintaining intracranial pressure above arterial systolic pressure. Contractile performance was determined with sonomicrometers arrayed to measure wall thickness in the anterior and posterolateral left ventricle. Brief aortic constrictions enabled derivation of the end-systolic pressure-thickness relationship, a relatively load-insensitive index of contractility. MEASUREMENTS AND MAIN RESULTS: No statistically significant hemodynamic differences were detected between the sham and time control groups. The brain-dead group displayed marked hemodynamic deterioration; 4 hrs after brain death was induced, mean arterial blood pressure, cardiac index, and the peak positive first derivative of left ventricular pressure were 63%, 35%, and 53% lower, respectively, than those values in the two control groups. However, the slope of the end systolic pressure thickness relationship remained at 92 +/- 12% of baseline levels 4 hrs after brain death was induced and was not significantly different from the two control groups. CONCLUSIONS: The hemodynamic deterioration after the induction of brain death could not be attributed to a decrease of myocardial contractility as measured by the end systolic pressure-thickness relationship. PMID- 8222692 TI - Effect of intravenous lipid emulsions enriched with gamma-linolenic acid on plasma n-6 fatty acids and prostaglandin biosynthesis after burn and endotoxin injury in rats. AB - OBJECTIVE: To study the effect of intravenous lipid emulsions enriched with gamma linolenic acid on plasma fatty acids and series-2 prostaglandins to determine if the slow conversion of linoleic acid by delta-6-desaturase to gamma-linolenic acid could be bypassed to provide substrate for the formation of dihomo-gamma linolenic acid, the immediate precursor for series-1 prostaglandins, in control and injured rats. Dihomo-gamma-linolenic acid can also compete with arachidonic acid for oxidative metabolism by cyclooxygenase to modulate series-2 prostaglandin biosynthesis. DESIGN: Prospective, randomized, controlled, double blind study. SETTING: Research laboratory at a university medical center. SUBJECTS: Thirty-three control and thirty-one injured male Sprague-Dawley rats were randomized into one of four parenteral dietary treatment groups. INTERVENTIONS: Rats were injured by the combined actions of a 30% body surface area full-thickness skin burn and a nonlethal injection of endotoxin (1 mg/kg ip). The rats were parenterally fed 200 kcal/kg/day, 1.5 g nitrogen/kg/day, and 30% of nonprotein calories as lipid (20% soybean lipid emulsion enriched with 2.7%, 4.4%, or 6.1% gamma-linolenic acid derived from borage oil) for 3 days. Control rats were treated similarly but were not injured. A 20% soybean/safflower oil lipid emulsion was used as the control diet (0% gamma-linolenic acid). Plasma was analyzed on day 3 to determine the concentrations of total fatty acids, thromboxane B2, 6-keto-prostaglandin F1 alpha, and bicyclo-prostaglandin E. MEASUREMENTS AND MAIN RESULTS: Parenteral nutrition with 2.7%, 4.4%, and 6.1% gamma-linolenic acid increased the plasma percentages (mol%) of gamma-linolenic acid and dihomo-gamma-linolenic acid in a dose-dependent fashion in control and injured rats. Supplementation with gamma-linolenic acid did not increase the plasma percentage of arachidonic acid as compared with the 0% gamma-linolenic acid lipid emulsion in control and injured rats. The ratio of dihomo-gamma linolenic acid to arachidonic acid was significantly increased in response to 4.4% and 6.1% gamma-linolenic acid in both the control and injured groups. The plasma ratio of thromboxane B2 to 6-keto-prostaglandin F1 alpha was substantially reduced with gamma-linolenic acid compared with 0% gamma-linolenic acid in injured rats. Bicyclo-prostaglandin E concentration was significantly higher with 2.7% gamma-linolenic acid in injured rats. Injured rats were protein catabolic, as evidenced by a net negative nitrogen balance and loss of body mass compared with controls, but neither group showed overt signs of intolerance to the diets. CONCLUSIONS: Supplementation of parenteral nutrition with gamma-linolenic acid had the following effects: a) increased plasma gamma-linolenic acid, dihomo-gamma linolenic acid, and bicyclo-prostaglandin E; b) increased the plasma ratio of dihomo-gamma-linolenic acid to arachidonic acid; and c) favorably reduced the ratio of thromboxane B2 to 6-keto-prostaglandin F1 alpha in injured rats. These results reflect the potential capacity of gamma-linolenic acid-enriched lipid emulsions to have the following actions: a) to increase dihomo-gamma-linolenic acid, which is the fatty acid precursor of the antiaggregatory, anti-inflammatory eicosanoid, prostaglandin E1; and b) to modulate arachidonic acid-derived series 2 prostaglandins after injury. PMID- 8222693 TI - Endotoxin infusion primes elicited neutrophils and Kupffer cells for platelet activating factor-induced and tripeptide formyl-methionine-leucine-phenylalanine induced basal free intracellular calcium concentration responses. AB - OBJECTIVE: To determine whether in vivo infusions of bacterial endotoxin prime rat Kupffer cells and elicited neutrophils by altering the basal free intracellular calcium concentration or the response of free intracellular calcium to platelet-activating factor or the tripeptide formyl-methionine-leucine phenylalanine (f-met-leu-phe). DESIGN: Prospective, randomized, controlled study. SETTING: Laboratory of a university medical school. SUBJECTS: Adult male Sprague Dawley rats. INTERVENTIONS: Rats were infused with sterile saline or with Escherichia coli endotoxin for 3 or 30 hrs via a venous catheter or an osmotic minipump. Rats were anaesthetized, and the livers were perfused in situ with a collagenase-containing buffer. Kupffer cells and elicited neutrophils were isolated by centrifugal elutriation, and were partially purified over Ficoll gradients. MEASUREMENTS AND MAIN RESULTS: Basal free intracellular calcium concentrations were determined with the calcium indicator, fluo-3. Various concentrations of platelet-activating factor or f-met-leu-phe were then added, and any alterations in free intracellular calcium values were quantified. Neither acute (3-hr) nor chronic (30-hr) infusions of endotoxin altered basal free intracellular calcium values. F-met-leu-phe-induced increases in free intracellular calcium concentrations were positively correlated with the percentage of neutrophils in Kupffer cell fractions. Three-hour infusions of endotoxin, which caused a significant inclusion of neutrophils in the Kupffer cell fraction, resulted in an enhanced response to f-met-leu-phe. This response was considered to be primarily due to the presence of neutrophils, because preparations that were > 95% Kupffer cells did not respond to f-met-leu-phe. Platelet-activating factor-induced increases in free intracellular calcium concentrations were also positively correlated with the percentage of neutrophils. However, Kupffer cells isolated from rats infused for 30 hrs with endotoxin contained < 12% neutrophils, and exhibited a ten-fold greater response to platelet-activating factor than did Kupffer cells isolated from saline-infused rats. CONCLUSIONS: In vivo infusions of endotoxin result in an enhanced response to platelet-activating factor by rat Kupffer cells. F-met-leu-phe does not appear to induce an increase in free intracellular calcium values in rat Kupffer cells, but it appears to do so in endotoxin-elicited neutrophils. These observations indicate that changes in calcium homeostasis may be one mechanism by which endotoxin primes Kupffer cells and elicited neutrophils for enhanced production of superoxide anion and/or nitric oxide. PMID- 8222694 TI - Effects of hemorrhage, hypoxia, and intravascular coagulation on bacterial clearance and translocation. AB - OBJECTIVE: This study was undertaken to discover if impaired blood clearance functions and killing capacity of the reticuloendothelial system contribute to the high occurrence rate of septic complications after shock, trauma, and thermal injury. DESIGN: Prospective, randomized, controlled trial. SETTING: Experimental laboratory in a university teaching hospital. SUBJECTS: Thirty-three standard breed rabbits of either sex. INTERVENTIONS: Defined numbers of Escherichia coli (1.3 x 10(8)) colony-forming units were injected intravenously 60 min after induction of hypoxia, standardized by defined reduction of oxygen uptake (60% to 75% of baseline) induced by hypoventilation (n = 6) or hemorrhage (n = 6), after complete defibrination caused by the snake toxin, ancrod (n = 6), and after 60 mins without intervention (controls, n = 6). At 180 mins after bacterial injection, the animals were killed and tissue samples of liver, kidney, spleen, and lung were collected for microbiological examinations. MEASUREMENTS AND MAIN RESULTS: Bacterial elimination from the blood and distribution pattern of viable bacteria in liver, spleen, kidney, and lung were investigated in hemorrhagic, hypoxic, and defibrinated rabbits. Compared with controls, there was a distinct alteration of the elimination kinetics of bacteria from the circulating blood in the experimental groups. First, the initial counts of viable E. coli were up to 300% (p < .05) higher in the defibrination, hemorrhage, and hypoxia groups than in controls. Second, greater numbers of E. coli were found in the blood for a significantly (p < .001) longer period, coupled with up to four times higher counts in organ homogenates in the hemorrhagic and defibrinated groups (p < .01) and more than 100 times higher counts than control values in the hypoxic animals (p < .001). CONCLUSION: Hemorrhage, hypoxia, and intravascular coagulation induce impaired bacterial clearance from the blood that is associated with altered organ distribution patterns, thus reflecting dysfunction of the reticuloendothelial system. PMID- 8222695 TI - Comparison of the acid-base status of blood obtained from intraosseous and central venous sites during steady- and low-flow states. AB - OBJECTIVE: To compare the acid-base status of blood obtained from the tibial intraosseous site with that status obtained from a central venous site during stead- and low-flow states in a piglet model. DESIGN: A prospective, observational study. SETTING: Animal laboratory at a university medical center. SUBJECTS: Nine 2-day-old piglets. INTERVENTIONS: Animals were anesthetized, intubated, and mechanically ventilated. A thermodilution pulmonary artery catheter was inserted via the right internal jugular vein and directed into the pulmonary artery. An arterial catheter was inserted into the right carotid artery and an intraosseous needle was inserted into the proximal tibial marrow cavity. Cardiorespiratory arrest was induced by discontinuation of ventilation. The animals were subsequently resuscitated by precordial compressions and ventilation. Blood samples were obtained from central venous and intraosseous sites during steady state and during resuscitation (low-flow state). RESULTS: No significant differences (p < .05) were found for pH, PCO2, and bicarbonate concentration when values that were obtained from the central venous and intraosseous sites were compared during steady- and low-flow states. CONCLUSIONS: The acid-base status of intraosseous blood is similar to that status of central venous blood. Intraosseous blood gas values may be an acceptable alternative to central venous blood gas values in judging central acid-base status during cardiopulmonary resuscitation. PMID- 8222696 TI - Free radicals and antioxidants in sepsis. AB - OBJECTIVES: The clinical condition of sepsis is caused by the release of numerous mediators from many cells. The purpose of this review is to describe the results of studies in which the role of free radicals and/or the potential therapeutic value of antioxidants are assessed. DATA SOURCES: The studies described in this review come from a variety of sources, including Med-Line CD-ROM computerized database, Index Medicus, and references identified from the bibliographies of pertinent articles and books. Reports were confined to English language articles from 1966 to 1992. STUDY SELECTION: All retrieved references in which free radical activity was assessed or antioxidants were measured or administered in sepsis or endotoxemia were included. This selection process encompassed clinical, animal and in vitro cell culture work. DATA EXTRACTION: Cited literature was found in reputable peer-reviewed clinical or basic science journals. DATA SYNTHESIS: Any contradictions in the results of studies are discussed. CONCLUSIONS: There is evidence that free radicals play an important role in the pathogenesis of sepsis. Antioxidant therapy has the potential to protect against such injury. It is suggested that combination therapy, which augments the endogenous antioxidant defenses, is likely to be the best approach. PMID- 8222697 TI - New technologies, critical care, and economic realities. PMID- 8222698 TI - The anencephalic organ donor: a challenge to existing moral and statutory laws. AB - OBJECTIVES: To inform physicians of the critical shortage of neonatal heart donors and to provide arguments for and against selecting brain-absent anencephalics as heart donors for brain-normal infants who are dying of hypoplastic left heart syndrome. DATA SOURCES: International scientific journals of medicine, genetics, epidemiology, bioethics, and public health; adjudicated U.S. civil court cases; and state regulations and statutes. STUDY SELECTION: Worldwide experience with anencephalics as homologous organ donors. DATA EXTRACTION: Demographic and epidemiologic data on anencephalic births and natural histories; U.S. civil, district, and appellate court case decisions directing or prohibiting organ donation; state determination-of-death acts; state uniform anatomical-gift acts. DATA SYNTHESIS: Organization of all data into either moral challenges or legal challenges to anencephalic organ donation. CONCLUSIONS: Statutory laws pose a greater challenge to anencephalic organ donation than moral laws. Case law reviews eliminate substituted judgment rulings in directing anencephalic organ donation. A redefinition of brain death applying only to human beings born without a brain would make more donor hearts available to brain normal infants dying of hypoplastic left heart syndrome. PMID- 8222699 TI - Use of anencephalic infants as organ donors. PMID- 8222700 TI - Severe pulmonary hypertension associated with shock and death in infants infected with Bordetella pertussis. PMID- 8222701 TI - A preprimed circuit stored for 45 days and used for emergency extracorporeal lung and heart assist. PMID- 8222702 TI - Death in the pediatric intensive care unit. PMID- 8222703 TI - Tumor necrosis factor and continuous arteriovenous hemodialysis. PMID- 8222704 TI - Prediction of need for physician as part of pediatric transport team. PMID- 8222705 TI - Neuromuscular blocking agents. PMID- 8222706 TI - Working with Jewish ultra-orthodox patients: guidelines for a culturally sensitive therapy. AB - The epistemological gap between the medical reality of mental health practitioners and the sacred reality of their Jewish ultra-orthodox patients poses a major challenge for therapy. Based on our work with psychiatric patients from the ultra-orthodox community of northern Jerusalem, we propose a set of guidelines to cope with this challenge. Basically, we seek to incorporate religiously congruent elements, composed of metaphoric images, narratives and actions, into the wide range of our "secular" treatment modalities in order to respond to the patient's suffering, often expressed through distinctively religious idioms of distress. This endeavor calls for "a temporary suspension of disbelief" on both sides. The guidelines presented include three sets of factors which appear pertinent to working with ultra-orthodox patients. The first set is contextual in nature, dealing with the image of the clinic and its physical setting; the second discusses the necessary role requisites of the therapists; and the third one, accorded a central importance, deals on various levels with the therapeutic interventions administered in terms of form and content. Several case vignettes are presented to illustrate three classes of religiously informed interventions: healing rituals, dream interpretation, and the use of culturally congruent metaphors and stories. In the concluding part we discuss ethical and instrumental issues that the proposed therapeutic guidelines may raise. PMID- 8222707 TI - Bedside manners: paradoxes of physician behavior in grand rounds. AB - This paper explains some of the routine of grand rounds in a major Israeli teaching hospital. It focuses on paradoxical behavior of physicians and others who participate in grand rounds. Behaviors are explained as boundary marking mechanisms meant to redefine statuses in the hospital and defend the physician from threats to his status. These paradoxical behaviors also function to permit the physician to enter and depart from the more culturally marginal territories of human experience, those which people try to deny or mask in everyday life. In particular, paradoxical behavior arises when dealing with death, waste, and sex. Paradoxical behavior is seen as a means of making a space in which everyday reality is masked and turned into play, creating a boundary between the medical reality and the social world. Paradox and other aspects of the grand rounds serve to restratify and counteract structural breakdown created by the physician's entry into states of pollution. PMID- 8222708 TI - Psychiatric intake as a mystery story. AB - The psychiatric intake process is a form of negotiated interaction, where patient presentations are affected by referral source, the illness itself, and illness behavior. At the same time, staff understanding of patient presentations is affected by institutional structures of the clinic, desire to control the clinical interaction, and broad developments in the psychiatric fields (especially the dominance of biopsychiatry). Much of the interplay involves patients withholding or reinterpreting information, staff imputations about both the withheld and the proffered information, and the resultant bargaining over what is at stake. Patients may simply be telling a story, but therapists are listening to it as a mystery story. They thus look for cues as to how things are signified and intended, and clues as to how this material fits together as a mystery. This leads to excessive reliance on a question-and-answer mode of engaging, staff interpretation of patient behavior and motivation, and control by staff of information. For patients, too, the clinical intake is a mystery in terms of what is wrong with them, what will be done for them, and how to conduct themselves in the intake. PMID- 8222709 TI - The cutting edge in organ transplantation. PMID- 8222710 TI - The cutting edge in trauma. PMID- 8222711 TI - The cutting edge in cardiovascular medicine. PMID- 8222712 TI - The cutting edge in neonatal critical care. PMID- 8222713 TI - The cutting edge in pediatric critical care. PMID- 8222714 TI - The cutting edge in burn care. PMID- 8222715 TI - The cutting edge in immunology. PMID- 8222716 TI - The cutting edge in pulmonary critical care. PMID- 8222717 TI - The cutting edge in neuroscience. PMID- 8222718 TI - The cutting edge in arrhythmias. PMID- 8222719 TI - Ascorbic acid regeneration by bovine iris-ciliary body. AB - Ocular tissues appear to require high levels of ascorbic acid and the elucidation of the mechanisms by which those tissues maintain the vitamin in its reduced state remains an important objective. The regeneration of ascorbate from its oxidative by-product, dehydroascorbic acid (DHAA), was studied in bovine iris ciliary body. Iris-ciliary body was removed by scalpel, weighed, minced, and homogenized in 20 mM MOPS, 62 mM sucrose, and 0.1 mM EDTA at pH 7.0. The homogenate was centrifuged and precipitated with ammonium sulfate such that maximal DHAA reducing activity was enhanced in a 50-75% ammonium sulfate fraction. This fraction was employed for subsequent characterization of DHAA reduction by iris-ciliary body. Results indicate that the iris-ciliary body enzymatically reduces DHAA to ascorbate at a rate significantly greater than can be accounted for by a nonenzymatic glutathione-dependent mechanism. In addition, saturation kinetics are observed, and the enzymatic activity is dependent on protein concentration, DHAA concentration, and reduced glutathione (GSH) concentration. The activity is sensitive to pH, to high temperature, and to digestion by trypsin and is greatest in the presence of both GSH and NADPH. The reducing activity is therefore attributed to one or more proteins that are distinct from the known ascorbate regenerating enzyme, GSH-dependent DHAA reductase (EC 1.8.5.1). PMID- 8222720 TI - Antimicrobial activity and in vitro corneal epithelial toxicity of antimicrobial agents for gram-positive corneal pathogens. AB - We assessed in vitro the antimicrobial activity of four agents (vancomycin, teicoplanin, mupirocin, and imipenem) which are effective against Gram-positive cocci causally associated with bacterial keratitis, as well as their corneal epithelial cytotoxicity. Minimal inhibitory concentrations inhibiting 90% of strains (MIC90s) against 10 strains each of methicillin-susceptible and resistant Staphylococcus aureus and Staphylococcus epidermidis, penicillin susceptible and -resistant Streptococcus pneumoniae, and viridans group streptococci were as follows: vancomycin (MIC90s 0.25-2 micrograms/ml), teicoplanin (MIC90s 0.25-4 micrograms/ml, mupirocin (MIC90s 0.12-4 micrograms/ml), and imipenem (MIC90s 0.008-0.25 micrograms/ml, except for methicillin-resistant staphylococci with MIC90 of 16 micrograms/ml). Cytotoxicity was assayed by uptake of 3H-thymidine by rabbit corneal epithelial cell cultures at drug concentrations of 12.5-100 mg/ml for vancomycin and teicoplanin, 1-8 mg/ml for mupirocin and 0.125-8 mg/ml for imipenem, with exposure times of 5, 30 and 60 min. Cytotoxicity was as follows: imipenem < mupirocin < vancomycin < or = teicoplanin. Resistance to methicillin-resistant S. aureus and S. epidermidis for imipenem and resistance to S. epidermidis and cytotoxicity for teicoplanin limit their consideration for suspected Gram-positive keratitis. On the other hand, vancomycin and mupirocin, because of their excellent therapeutic indices, should be considered for this indication. PMID- 8222721 TI - A comparison of two methods of measuring aqueous flow in humans: fluorophotometry and flare measurement. AB - The technique of Anjou and Krakau for measuring flare and Krakau's method for estimating aqueous humor flow in humans was compared to the fluorophotometric procedure of Jones and Maurice. The precision of flare was measured in 20 normal human volunteers, and the precision of flow was measured with fluorophotometry in 24 volunteers. In addition, the circadian rhythms of both flow and flare were measured simultaneously in a separate group of 20 subjects. This simultaneous measurement allowed us to calculate the entry rate of scattering substances into the aqueous humor as a function of time of day. This entry rate is critical for measuring flow by the method of Krakau. The mean coefficient of variation (+/- SD) of repeated measurements of flare at the same time of day was 23.2% +/- 10.3%. The mean coefficient of variation of the measurement of aqueous flow by fluorophotometry was 15.9% +/- 8.2%, significantly better than the measurement of flare (p = 0.01). The daytime entry rate of scattering substances into the aqueous humor varied from 1.39 +/- 0.86 microgram/min (calculated as albumin) to 1.05 +/- 0.58 microgram/min. The rates in daytime hours were not significantly different from each other. At night during sleep, the entry rate was 0.67 +/- 0.48 microgram/min, significantly slower (p between 0.002 and < 0.001). These data indicate that the flare technique cannot be used alone to calculate the circadian rhythm of flow and that direct measurement of flow by fluorescein clearance is likely to be the most repeatable and accurate of the two. PMID- 8222722 TI - Allotransplantation of rabbit retinal pigment epithelial cells double-labelled with 5-bromodeoxyuridine (BrdU) and natural pigment. AB - To facilitate identification of transplanted retinal pigment epithelial (RPE) cells, we sought to double-label the cells with 5-bromodeoxyuridine (BrdU) and with natural pigment. The BrdU is not lost during cell division but does require immunohistochemical methods for visualization; the pigment, on the other hand, allows immediate, obvious identification, but is gradually lost with cell division. Together they provide a convenient, long-term double label. Herein we report the successful allotransplantation of double-labelled RPE cells onto Bruch's membrane of albino rabbits. The labelled cells were localized by anti BrdU antibody and the avidin biotin-alkaline phosphatase complex (ABC-AP) method, and by visible inclusions of pigment. Using this double-label method, allotransplanted RPE cells could be readily and reliably identified on the recipient Bruch's membrane eight months after transplantation. The cells had distinct basal and apical morphology, and were in close contact with the photoreceptor outer segments of the host. This successful allotransplantation raises the possibility that the subretinal space of the rabbit might enjoy some degree of immunologic privilege. PMID- 8222723 TI - Beta-cyclodextrins enhance bioavailability of pilocarpine. AB - Cyclodextrins have been used to improve drug solubility, stability and absorption for oral and parenteral administration. However, their potential for improving ocular drug delivery has received little attention. To evaluate the ability of hydroxypropyl-beta-cyclodextrins to improve ophthalmic drug bioavailability following topical administration, the miotic effect of topical solutions of pilocarpine was studied in New Zealand White rabbits. Pilocarpine varying in dose from 5 to 500 micrograms in the presence or absence of 5% cyclodextrin was administered (50 microliters) topically and the change in pupil diameter determined. These results demonstrated that pilocarpine alone or in the presence of cyclodextrin produces a dose-related reduction in pupil diameter. The addition of cyclodextrins produced a significant left-shift in the dose response curve, with an ED50 of 64 micrograms and 19 micrograms for pilocarpine and pilocarpine/5% cyclodextrin solutions, respectively. Studies in which the concentration of cyclodextrin was varied revealed that a one-to-one molar ratio of pilocarpine to cyclodextrin was sufficient to provide maximum increase in pilocarpine bioavailability. Electrophysiology and scanning electron microscopic studies demonstrated that cyclodextrin does not disrupt the normal ion transport currents, barrier properties or surface features of the corneal epithelium. Viscosity measurements indicated that difference in the viscosity of pilocarpine and pilocarpine/cyclodextrin solutions cannot account for increased bioavailability of pilocarpine. These data support the idea that the addition of cyclodextrin significantly improves the ocular bioavailability of pilocarpine. This enhanced bioavailability of pilocarpine does not appear to be due to a mechanism destructive to the epithelium or to an increase in vehicle viscosity. PMID- 8222724 TI - Ocular inflammatory effects of intravitreally injected interleukin-2. AB - The primary, known physiologic effect of interleukin-2 (IL-2) is to act as a T lymphocyte growth factor. We investigated the potential contribution of IL-2 to intraocular inflammation by studying the inflammation resulting from the intravitreal injection of recombinant, human IL-2 in New Zealand white rabbits. Serial slit lamp observations indicated that 40 microgram of intravitreally injected IL-2 induced an anterior uveitis which was maximal 5 days after the injection. Inflammation was less marked but still significant with amounts of IL 2 as low as 400 ng. Direct examination of aqueous humor confirmed elevations of protein, prostaglandin E2, and mononuclear cells which correlated with the clinical observations. The kinetics of the response to intravitreal IL-2 distinguished it from the responses to other intravitreally injected cytokines such as interleukins 1, 6, or 8 as well as tumor necrosis factor. Intramuscular injection of cyclosporine A significantly reduced the protein extravasation associated with IL-2 injection, but cyclosporine had no effect on inflammation secondary to an intravitreal injection of interleukin-1. These observations implicate IL-2 as a potential contributor to uveitis. In addition, the studies with cyclosporine indicate the heterogeneity of inflammation such that pharmacologic agents which affect one cause of uveitis are not necessarily efficacious in another model. PMID- 8222725 TI - The platelet-activating factor precursor of the injured cornea is selectively implicated in arachidonate and eicosanoid release. AB - The purpose of this study was to isolate the platelet-activating factor (PAF) precursor and other choline phosphoglycerides (GPC) i.e. the alkenylacyl and diacyl lipids from the rabbit cornea, to analyze their fatty acid content and to determine which pool was the most susceptible to arachidonate depletion when activated corneal tissue released arachidonic acid (AA) and metabolites. Rabbit iridal GPC was also analyzed for comparative purposes. The fatty acid methyl esters of the GPC components extracted from the rabbit cornea and iris-ciliary body, isolated by high performance liquid chromatography (HPLC), were determined by capillary gas liquid chromatography. Rabbit corneas were labelled in vivo by intracameral injection of 3H-AA (1 microCi, specific activity = 218 Ci/mmol) and cryogenically injured 18 h later. Corneas were incubated in vitro and the AA and eicosanoids released into the medium were extracted and separated by HPLC. The GPC was extracted from the tissues and the labeling of the three GPC constituents was quantified by liquid scintillation counting. The corneal and iridal PAF precursor represented 4.1 +/- 0.2% and 2.9 +/- 0.2% respectively of total GPC in those tissues. On a mole basis, the alkyl arachidonoyl species constituted 12.7 +/- 0.7% of the corneal and 38 +/- 0.6% of the iridal PAF precursors respectively. The release of AA and prostaglandins by the cornea was linear until 15 min; whereas 12-HETE levels continuously increased until 60 min. All GPC components lost label but 1-O-alkyl-2-arachidonoyl was the most affected, with its labeled content 50% less than the non-injured control.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222726 TI - The effect of serotonin on the rabbit isolated iris sphincter muscle. AB - The rabbit isolated iris sphincter muscle maintained in an isotonic state is unaffected by applied serotonin (5-hydroxytryptamine or 5-HT) whereas carbachol causes the muscle to contract. Serotonin does, however, produce a relaxation of the contracted muscle in a dose-dependent manner. This effect is also induced by the 5-HT receptor agonists 8-OH-DPAT (8-hydroxy-2-[di-n-propyl-amino] tetralin, RU 24969 (5-methoxy-3-[1,2,3,6, tetrahydro-4-pyridinyl]-1-indole) and ipsapirone, suggesting the involvement of 5-HT1A receptors. This view is supported by the finding that metergoline, methysergide and propranolol all counteracted the effect produced by serotonin. While 5-HT3 receptors are not involved in the described process, a minor involvement of 5-HT2 receptors cannot be excluded as methysergide partially counteracted the serotonin response. These data provide evidence that serotonin receptors, in particular the 5-HT1A subtype, may be associated with the iris sphincter muscle and suggest their involvement in the regulation of pupil size. PMID- 8222727 TI - Elemental studies in rat lens during galactose cataract reversal. AB - Alterations in elemental composition of the normal lens have been reported to accompany galactose cataract development in rats. In this report we present the changes in regional distribution of Na, K, Cl, P, S and Ca during the reversal of galactose-induced cataracts. Elemental X-ray maps of lenses from young female Sprague Dawley rats fed 50% galactose for 20 days were examined at 0, 20, 40 and 90 days following the transfer of galactose fed rats to Purina Rat Chow diet. Reinstatement of normal elemental distribution accompanied the progression of lens transparency. By 90 days on the rat chow diet, K had increased and Na, Cl and Ca had decreased so that a near normal lenticular distribution of these elements was established. The reinstatement of elemental distribution during cataract reversal followed a pattern similar to that observed for alterations during cataract development, initiating near the equatorial surface and expanding centrally. The correlation between the alterations in the distribution of the elements studied and our previously reported morphological investigation of lenses during galactose cataract reversal is discussed in this report. PMID- 8222728 TI - Age-related changes of sulfated proteoglycans in the human lamina cribrosa. AB - Sulfated proteoglycans in the lamina cribrosa of the optic nerve head from individuals aged 2 months, 18 months, and 23, 35, 44, 55, 67, 74, and 88 years were studied by electron microscopy after cuprolinic blue dye binding. Within the cores of the laminar plates, cuprolinic blue-positive chondroitin/dermatan sulfate proteoglycan filaments of different sizes were found associated with collagen fibers. In addition, small punctate and filamentous structures that represented heparan sulfate proteoglycan molecules were associated with the basal laminae of astrocytes and blood vessels. In the eyes of older individuals, the chondroitin/dermatan sulfate and heparan sulfate proteoglycan filaments were found to be shorter than those in younger persons. A mild decline with aging in the diameter of the filaments was also noted. Our findings illustrate the age related changes in the proteoglycans in the human lamina cribrosa, which may help explain why the optic nerve head is more susceptible to damage with aging. PMID- 8222729 TI - Cultured ocular cells and extracellular matrices: role of growth factors, retinoic acid and cell polarity. AB - Culture of various types of cells on gelled, reconstituted extracellular matrices results in decreased cellular proliferation. In the present study, we evaluated several possible mechanisms for this inhibition, as applied to cultured bovine retinal microvascular endothelial cells (EC) or to retinal pigment epithelial (RPE) cells: whether the inhibition might be related to (a) inactivation of fibroblast growth factor (FGF) by binding of the molecules present in the medium to a matrix component; (b) release of an inhibitor by the matrix in culture; or (c) inhibitory properties of the matrix macromolecules themselves. Our results suggest that mechanism (c) is most likely. The reasons are, first, that culture of EC or RPE cells on several different extracellular matrix substrates in the presence of logarithmically increasing concentrations of acidic or basic fibroblast growth factors (aFGF or bFGF) leads to a vertical shift of the plots of cell number after 4 days in culture vs. log growth factor concentration for both types of cells. The same result obtains when cells are cultured with logarithmically increasing concentrations of all-trans retinoic acid, which inhibits EC but not RPE cell proliferation in a dose-dependent fashion. This is consistent with mechanism (b) or (c), but not (a), for which one would expect a horizontal shift. Second, washing the matrices prior to the plating of cells with 1M NaCl, which elutes aFGF and partially elutes bFGF molecules from basement membranes, does not alter the growth of cells plated after the wash. This suggests also that growth factor binding to the matrix is not a likely mechanism for the observed inhibition. Incubation of matrices with culture medium prior to plating cells does not usually alter the ability of the medium thus "conditioned" to support cell growth, arguing against the possibility that the matrices release a soluble activator or inhibitor of such growth. However, in some experiments performed with lots of Matrigel (a commercially available basement membrane extract from a murine tumor) obtained prior to mid-1991, media "conditioned" by incubation with this matrix did show enhanced ability to facilitate EC and RPE cell proliferation. Finally, if RPE cells or EC are plated on various substrates, allowed to attach for 24 hr., and then the same or other substrates are poured over the cells, the effect on proliferation of the matrices plated on the apical surfaces of the cells is often less than that of matrices plated adjacent to their basal surfaces. Although in most cases these differences are not statistically significant, there is an apparent trend.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8222730 TI - Transforming growth factor-beta stimulates collagen and fibronectin synthesis by human corneal stromal fibroblasts in vitro. AB - The effects of transforming growth factor-beta (TGF beta) and epidermal growth factor (EGF) on the synthesis of collagen and fibronectin, and on the proliferation of human corneal stromal fibroblasts in vitro, were evaluated. Human corneal stromal fibroblasts in culture were incubated for 48 hours with TGF beta or EGF in the absence of serum. Collagen and fibronectin in the culture media were measured by a collagenase-digestion assay and a competitive ELISA, respectively. The effects of the growth factors on proliferation were assessed by 3H-thymidine incorporation. Collagen synthesis was dose-dependently stimulated by TGF beta; at a concentration of 1 ng/ml of TGF beta, a 120% increase in collagen synthesis was seen over that of controls (p < 0.01). EGF, at a concentration of 10 ng/ml, induced a 40% increase in collagen synthesis over that of controls (p < 0.01). The maximum stimulation by TGF beta was greater than that by EGF (p < 0.05). Fibronectin synthesis was stimulated by TGF beta and EGF in a dose dependent manner; 230% (p < 0.001) and 210% (p < 0.01) increases in fibronectin synthesis were caused by 10 ng/ml TGF beta and EGF, respectively. TGF beta and EGF dose-dependently stimulated 3H-thymidine incorporation. The maximum increases in 3H-thymidine incorporation reached 180% (p < 0.001) and 190% (p < 0.001) over that in controls, at 10 ng/ml concentrations of TGF beta and EGF, respectively. In conclusion, both TGF beta and EGF are potent stimulants of collagen and fibronectin synthesis and proliferation. Therefore, these two growth factors may be effective alternatives or additional choices for the treatment of corneal ulcer. PMID- 8222731 TI - Opsin synthesis in the C57BL/6-mivit/mivit mouse model of retinal degeneration. AB - The capacity of photoreceptor cells to synthesize opsin was evaluated in a newly described mouse model of retinal degeneration, the C57BL/6-mivit/mivit. The mivit/mivit mouse loses photoreceptor cells at a rate of about one row per month beginning at 8 weeks, ROS are severely disrupted at 4 months, RPE is unevenly pigmented. Retinas of affected and control mice ages 4, 6, 8, 12, 16, 20, 24, 28, 32 and 52 weeks were incubated for 2 hours in medium containing [3H] leucine. Homogenates of retina samples were subjected to SDS-PAGE using disc gels. The gels were sliced and counted by scintillation. The incorporation of [3H] leucine into opsin was compared with its incorporation into other retinal proteins. During the early time points studied, mivit/mivit retinas incorporated proportionately similar amounts of [3H] leucine into opsin versus other retinal proteins as did controls. At 12 weeks, the percentage was about 80% and it continued to decline over the succeeding weeks studied. By 1 year, the proportion of leucine incorporated into opsin versus other proteins was only about 23% the amount incorporated in controls. The results of the present study suggest that the mivit/mivit photoreceptor cells are able to synthesize opsin and the gradual decline in synthetic ability follows the gradual loss of cells and is not correlated with the disruption of ROS. PMID- 8222732 TI - Photoreceptor-specific protein expression of mouse retina in organ culture and retardation of rd degeneration in vitro by a combination of basic fibroblast and nerve growth factors. AB - Previously we have presented the morphological features of a neonatal mouse retinal explant kept in culture for 3 to 4 weeks. To further evaluate the organotypic parameters of the tissue we have examined the presence of opsin, S antigen, and interphotoreceptor retinoid-binding protein (IRBP) in the same experimental paradigm, using light microscopic immunocytochemistry. In vitro, opsin and S-antigen staining is found in photoreceptor somata from genetically normal explants and those derived from mice with the rd or the rds mutation. When present, inner and outer segments label more intensely. No IRBP staining has been found in cell bodies of any genotype. However, some labeling is found in the plexiform layers and in the inner segments. The results indicate that photoreceptor proteins are continuously produced in vitro. This further establishes the organotypic nature of the retinal explant in culture. The administration of growth factors to these explants has been investigated. Neither basic fibroblast growth factor nor nerve growth factor alone has affected the explants phenotype. However, the combination of these proteins has significantly retarded rd cell loss in vitro. PMID- 8222733 TI - Light evoked inositol trisphosphate release in the rat retina in vitro. AB - Light exposure not only elicits a photic response but may also alter the metabolism and functional properties of the retina. This may be evoked by the stimulation of phospholipid derived second messenger systems. In this study, we investigated the light-evoked release of inositol 1,4,5-triphosphate in the isolated rat retina in vitro by means of high performance liquid chromatography (HPLC) detection. After prelabelling of isolated retinae with tritiated myo inositol in darkness, they were exposed to no light or to white fluorescent light of 10,000 lux illuminance for 3,5 and 10 sec, respectively. We observed a 200% increase in the release of inositol 1,4,5-trisphosphate compared to basal values in darkness after 3 sec of light exposure with a decline after 5 sec and a return to basal values after 10 sec indicating a rapid breakdown of inositol 1,4,5 trisphosphate. Our data confirm previous studies in the amphibian retina and photoreceptors and demonstrate for the first time a light evoked inositol 1,4,5 trisphosphate release in the mammalian retina. PMID- 8222734 TI - Effects of antimicrobials on corneal epithelial migration. AB - The slowed healing rates observed by some investigators may be caused by vehicles or preservatives in the antimicrobials preparations tested. To determine whether antimicrobials directly inhibit corneal epithelial wound healing, we cultured blocks of the rabbit cornea in media containing various concentrations of antibiotics or antimicrobials (at 1, 10, or 100 micrograms/ml); after 24 hours, we measured the distance of epithelium that had migrated down the side of each block. The higher concentrations of fluoroquinolones (ofloxacin; 74 +/- 5.8% of control at 100 micrograms/ml, p < 0.05, ciprofloxacin; 4.4 +/- 1.5% of control at 100 micrograms/ml, p < 0.01, or norfloxacin; 71 +/- 7.0% at 10 mu g/ml, p < 0.01, and 1.5 +/- 0.4% of control at 100 mu g/ml, p < 0.01) and the highest concentrations of peptides (polymyxin B; 64 +/- 3.0% of control at 100 micrograms/ml, p < 0.01, or colistin; 67 +/- 5.7% of control at 100 micrograms/ml, p < 0.01) or fosfomycin (79 +/- 6.2% of control at 100 micrograms/ml, p < 0.05) had an inhibitory effect on corneal epithelial migration. Among aminoglycosides tested, sisomicin (85 +/- 10.0% of control, not significant), dibekacin (76 +/- 11.6% of control, p < 0.05) and streptomycin (77 +/- 9.4% of control, not significant) were inhibitory at 100 micrograms/ml, but tobramycin had no effect. Penicillins (aspoxicillin, sulbenicillin or ampicillin), cephalosporins (cefmenoxime or cefminox), oxytetracycline, erythromycin and chloramphenicol did not affect epithelial migration at all. These results demonstrate that some antimicrobials are inhibitory at high concentrations, but penicillins, cephalosporins, oxytetracycline, erythromycin or chloramphenicol has no inhibitory effect on corneal epithelial migration. PMID- 8222735 TI - Detection and partial characterization of ocular cicatricial pemphigoid antigens on COLO and SCaBER tumor cell lines. AB - Ocular cicatricial pemphigoid (OCP) is a chronic autoimmune inflammatory disease which affects the conjunctiva and other squamous epithelial mucous membranes resulting in a scarring process. It is characterized by the deposition of an anti basement membrane zone (BMZ) antibody in vivo. Sera from 11 patients with active OCP were studied. Using monkey esophagus and normal skin as substrate, weak staining of the BMZ was observed in conventional indirect immunofluorescence (IIF) assay. Using salt split human skin as substrate, the OCP sera demonstrated binding to the epidermal side of the split, in low titers with weak staining. Ten of the 11 sera were positive on an immunoblot assay using COLO and SCaBER tumor cell lysates demonstrating 230, 205, 160, and 85 kD proteins. Sera from six bullous pemphigoid (BP) patients, with only cutaneous involvement and high titer of anti-BMZ antibody, as detected by IIF, also bound to 230, 160, and 85 kD proteins on both lysates in comigration experiments. Serum from five normal individuals and two patients each with severe atopic conjunctival disease, erythema multiforme with chronic conjunctivitis and systemic lupus erythematosus (SLE), did not demonstrate those bands. When the two lysates were first absorbed with BP sera and then the same lysates were immunoblotted with OCP sera, in all ten OCP sera the 230, 160, and 85 kD bands were eliminated and only a single 205 kD band was uniformly present. These results indicate that OCP sera recognize peptide(s) present in 230, 205 and 160 kD proteins in lysates from COLO and SCaBER tumor cells. These proteins contain the immunodominant region of the BMZ molecule(s) in which the OCP antigen(s) reside. The OCP antigen(s) appears to be distinct from the BP antigen(s). PMID- 8222736 TI - Distribution of endothelin-like immunoreactivity in the human ciliary epithelium. AB - In primate eyes the injection of Endothelin (ET) 1 into the anterior chamber lowers the intraocular pressure by increasing outflow facility. A possible source for aqueous ET is the ciliary epithelium, since virus transformed human nonpigmented ciliary epithelial cells secrete ET into the medium and immunohistochemically stain with antibodies against ET 1. This study demonstrates that human ciliary epithelium exhibits positive endothelin-like immunoreactivity with antibodies against ET 1. There are, however, regional differences with regard to the staining intensity. Pronounced staining was observed in the nonpigmented and pigmented epithelium at the crests of the pars plicata. In the valleys of the pars plicata and in the pars plana no immunolabeling was found. PMID- 8222737 TI - Identification of receptor tyrosine kinases in the embryonic chicken lens. AB - Protein phosphorylation plays a critical role in the control of growth and regulation of many eukaryotic cells. Members of the protein tyrosine kinase (PTK) family of peptides function as growth factor receptors and oncoproteins. A common feature of members of the PTK family is a highly conserved intracellular catalytic domain. We analyzed the chicken lens epithelium, which responds to several known growth factors, for the presence of receptor PTK's. Using reverse transcription polymerase chain reaction (rtPCR) and degenerate primers made to conserved regions within kinase domains, we amplified RNA from embryonic day 6 (E6) lens epithelium and sequenced 135 cDNA clones. Sixteen distinct kinase sequences were obtained. Eight of these sequences represented kinase domains of known mammalian growth factor receptors, and six represented intercellular kinases. Two sequences appeared to code for new kinases. The amino acid identity of the chicken homologs ranged from 80-100% when compared to their mammalian counterparts. PMID- 8222738 TI - An improved ocular perfusion system. AB - We describe the design, construction and testing of an improved ocular perfusion system for outflow facility measurement. The essential feature of the system is the measurement of a differential pressure across a calibrated length of tubing, which provides a stable and accurate reading of facility and hence also of flowrate. Perfusion of calibrated lengths of tubing showed excellent accuracy, while perfusions of enucleated eyes showed very low noise levels. The system is relatively inexpensive, easy to construct and maintain, insensitive to vibrations and other noise, and accurate. It is therefore felt to be superior to conventional "hanging bucket" ocular perfusion systems. PMID- 8222739 TI - A double-blind comparison of oral ketoprofen 'controlled release' and indomethacin suppository in the treatment of rheumatoid arthritis with special regard to morning stiffness and pain on awakening. AB - A double-blind, double-dummy, crossover study was carried out in 8 centres to compare the efficacy and tolerability of 'controlled-release' ketoprofen tablets (200 mg) with that of indomethacin suppositories (100 mg) in out-patients with definite or classical rheumatoid arthritis. Patients were allocated at random to receive a daily bedtime dose of either 1 ketoprofen tablet or 1 indomethacin suppository plus the dummy of the other formulation for a period of 3 weeks. They were then crossed over to the alternative treatment for a further 3 weeks. Daily diary records were kept by patients of the number of night-time awakenings due to pain, pain severity at awakening in the morning and the duration of early morning stiffness. Treatment efficacy was also assessed at the end of each trial period by means of an articular index and by physician's and patient's overall evaluation of response. Adverse effects spontaneously mentioned by the patients or elicited by direct questioning using a symptom check-list were recorded. Statistical analysis of the results from 83 evaluable patients showed that the 'controlled-release' tablet formulation of 200 mg ketoprofen was equally as effective as the 100 mg indomethacin suppository in the treatment of rheumatoid arthritis, especially with regard to pain at awakening and morning stiffness. Side-effects in both groups were those commonly seen with non-steroidal anti inflammatory drugs and, as expected, gastro-intestinal and CNS disturbances predominated. Overall, side-effects were fewer with ketoprofen than with indomethacin. PMID- 8222740 TI - Effects on fibrinolytic activity of corn oil and a fish oil preparation enriched with omega-3-polyunsaturated fatty acids in a long-term study. AB - In addition to their beneficial effects in reducing platelet responsiveness, it has been a matter of controversy whether polyunsaturated fatty acids impair the fibrinolytic system or not. In a double-blind, parallel clinical trial, 40 subjects were randomized to treatment with 6 g/day of corn oil, or to 6 g/day of a fish oil preparation, enriched with omega-3-polyunsaturated fatty acids (2.0 g/day of omega-3-PUFA). Clinical and fibrinolytic variables were measured before and after 5 months of treatment. In the corn oil group, plasminogen activator inhibitor (PAI-1) decreased significantly but in the cod liver oil group, PAI-1 remained unchanged. Activities and mass concentrations of tissue plasminogen activator (t-PA) were unchanged in both groups. It is concluded that, in the doses given here, both these preparations have small or no effects on the fibrinolytic system. PMID- 8222741 TI - Halofantrine in the treatment of acute malaria: a multi-centre study in 268 patients. AB - The clinical efficacy and tolerability of halofantrine, a new antimalarial schizontocide, was studied in a multi-centre trial involving 268 patients ranging in age from 6 months to 58 years. The patients were suffering from acute uncomplicated malaria due to either P. vivax or P. falciparum. Patients were treated orally with 3 doses of halofantrine hydrochloride, 500 mg/6-hourly in adults or 8 mg/kg body weight 6-hourly in children. The overall cure rate was 96.9%. The mean fever clearance time for different species was as follows: P. vivax--39.1 hours, P. falciparum--43.2 hours, mixed infection--60.0 hours, and the mean parasitaemia clearance times were 47.7, 55.1 and 72.0 hours, respectively. Recrudescence was reported in 11 (4.1%) patients, although all of them were parasite-free on Day 7 post-treatment. No haematological or biochemical abnormalities were noted. The drug was very well tolerated and no significant side-effects were reported. Halofantrine was found to be highly effective in acute malaria and offers an important alternative to existing medications. PMID- 8222742 TI - Metoclopramide in the treatment of reflux oesophagitis: a comparison of normal and controlled-release formulations. AB - Twenty patients with reflux oesophagitis were recruited into a randomized, single (investigator)-blind, two-part crossover study of controlled-release metoclopramide ('Gastrobid Continus' tablets) 15 mg given twice daily, and normal release metoclopramide ('Maxolon'), 10 mg given 4-times daily. Both treatment regimens were efficacious in reducing the severity of heartburn, the incidence of waking at night due to pain from heartburn and the severity of gastro-intestinal symptoms. The incidence of volunteered and sought side-effects/symptoms was similar on both treatments. More patients preferred the controlled-release than the normal-release formulation. It is concluded that 15 mg controlled-release metoclopramide twice daily and 10 mg normal-release metoclopramide 4-times daily are equally effective and well-tolerated treatment regimens, but that the controlled-release formulation has the advantage of a smaller dose and twice daily administration. PMID- 8222743 TI - Controlled trial of a semi-rigid bandage ('Scotchrap') in patients with ankle ligament lesions. AB - A randomized, controlled, parallel-group study was performed to compare the clinical efficacy of a semi-rigid bandage ('Scotchrap') with the standard adhesive tape treatment for ankle ligament injuries. The semi-rigid bandage support wrap consists of a knitted fibreglass fabric impregnated with a polyurethane resin. The cotton stockinet and the synthetic cast padding are used for protection of pressure points and any bony prominences. Prior to application, the 'Scotchrap' must be soaked in water after which it is applied in a way similar to that used for a plaster of paris bandage. The finished bandage remains elastic, allowing normal dorsal and plantar flexion of the foot, but preventing inversion and eversion, and pronation and supination. At rest, the bandage always redresses the immobilized ankle into the neutral position. One hundred and thirty six patients were included in the study. The semi-rigid bandage group comprised 59 patients and the tape group had 57 patients. These two groups were statistically comparable. The intake questionnaire included a general and a specific anamnesis form as well as a physical examination form. Follow-up data were recorded every 2 weeks. After termination of the treatment, the patients evaluated treatment efficacy. Statistical analysis of 126 items per patient with chi 2-tests (p < or = 0.05) and Fisher's exact tests (p < or = 0.05) revealed no statistical significant differences in treatment result. The patient assessments of treatment efficacy of both types of bandage were also comparable. Both types of bandage gave sufficient stability (98%; p < 0.005) and were equally effective.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8222744 TI - Inhibitory effects of azelastine hydrochloride on Ca2+ influx, actin polymerization and release of eosinophil cationic protein of an eosinophilic leukaemia cell line EoL-1. AB - The inhibitory effects of azelastine hydrochloride on PAF-induced and fMLP induced Ca2+ influx, actin polymerization and calcium ionophore A23187-induced and aggregated IgG-induced release of eosinophil cationic protein (ECP) of an eosinophilic leukaemia cell line, EoL-1, were examined. EoL-1 cells cultured with 0.2 mM dibutyryladenosine-cyclic monophosphate for 48 hours showed an increase in intracellular free Ca2+ concentration ([Ca2+]i) and actin polymerization when stimulated by PAF and fMLP. Azelastine hydrochloride inhibited PAF-induced and fMLP-induced Ca2+ influx ([Ca2+]i) in a dose-dependent manner with an IC50 of 1 x 10(-8) M and 1 x 10(-7) M, respectively. It also inhibited PAF-induced and fMLP induced actin polymerization in a dose-dependent manner up to 40% and 30%, respectively. EoL-1 cells were differentiated to contain ECP in their eosinophilic granules when cultured for 9 days with supernatants of a human adult T cell leukaemia cell line, HIL-3 (HIL-3 sup). Calcium ionophore A23187 and aggregated IgG induced the secretion of ECP by EoL-1 cells. Azelastine hydrochloride inhibited the secretion of ECP in a dose-dependent manner. These inhibitory effects were seen even at therapeutic concentrations of 10(-8) M to 10(-9) M. These results indicate that the therapeutic effects of azelastine hydrochloride as an anti-allergic agent may include inhibition of the accumulation of eosinophils into the locus of allergic inflammation and of the release of cytotoxic granules from eosinophils. PMID- 8222745 TI - A post-marketing surveillance survey on the use of dipotassium clorazepate in clinical practice in the Philippines. AB - A post-marketing surveillance survey of dipotassium clorazepate usage was carried out in the Philippines between July 1990 and April 1991. Data were analyzed on 562 patients suffering from conditions in which anxiety was a major component. The results suggest that the medication was prescribed in accordance with the generally accepted best practice for benzodiazepines. Overall, 85% of patients were reported to have recovered and only 2% reported minor side-effects. Dipotassium clorazepate was thus confirmed to be a highly effective and relatively safe anxiolytic agent. PMID- 8222746 TI - Otitis externa: clinical comparison of local ciprofloxacin versus local oxytetracycline, polymyxin B, hydrocortisone combination treatment. AB - Thirty patients with otitis externa were included in a prospective, randomized, parallel group comparison study. Treatment regimens used were ciprofloxacin (2 mg/ml) as ear drops twice daily or drops containing oxytetracycline (5 mg/ml) polymyxin B (10,000 units/ml) and hydrocortisone (15 mg/ml) twice daily for 7 days. Evaluation was by clinical signs and symptoms and bacteriological findings from ear swabs prior to and 3 and 8 days after start of therapy, together with long-term follow-up if necessary. Sixteen patients were included in the ciprofloxacin and 14 in the combined antibiotic group. Six of 16 and 7 of 14 initial swabs, respectively, contained Pseudomonas aeruginosa. Mixed Gram negative and Gram-positive pathogens were found in 3 patients in the ciprofloxacin group. All Gram-negative isolates were sensitive to ciprofloxacin. Microbiological evaluation showed rapid eradication of all bacterial pathogens in the ciprofloxacin group with 1 persistence of Candida albicans but eradication of Staphylococcus aureus in the same ear with marked clinical improvement. There was 1 case of supercolonization in the ciprofloxacin group with Candida parapsilosis. In the combined antibiotic group, there were 7 persistencies of which 5 were P. aeruginosa. No adverse effects were noted in either group. Treatment was assessed as a complete success in 14/16 and 5/14 cases, partial success in 2/16 and 4/14 cases and unsuccessful or indeterminate in 0/16 and 5/14 cases, respectively. Ciprofloxacin thus appears to be a useful and effective agent in the local therapy of otitis externa. PMID- 8222747 TI - Cancer in the elderly. AB - This article reviews geriatric oncology and assesses options for treatment and care of the elderly patient with cancer. The size of the population over 65 years old is defined, with particular reference to the continuing growth of this subsection of the community. The high incidence of many cancers and their associated mortality rates in the elderly are identified and the epidemiology of such diseases in the geriatric population is addressed. Given the discrepancies in incidence and survival rates between patients younger and older than 65 years, the association between tumorigenesis and the aging process is explored. Specific aspects of tumor growth in the elderly are considered. General considerations of therapy for elderly patients with cancer are discussed, including the pharmacokinetics and pharmacodynamics of chemotherapy in those over 65 years old, surgical options, the use of radiotherapy, and overall patient assessment. Next, treatment options for individual cancer states are reviewed, with particular emphasis on newer treatment options designed specifically for the elderly. Sections on cancer screening and supportive care are also included, the latter dealing with aspects of symptom control, quality of life assessment, and the physical and psychologic rehabilitation of the elderly patient with cancer who is undergoing treatment. Conclusions are then drawn as to the extent of the oncological process in those over 65 years old, with particular emphasis on the underdiagnosis and undertreatment of many malignancies in the past. The challenge created by the growing elderly population is underscored and necessary plans of action for oncologists in the future are defined. Such proposals are necessary if inroads are to be made into the unacceptable morbidity and mortality rates borne by our elderly patients with cancer. PMID- 8222748 TI - Pharmacologic stress myocardial perfusion imaging. PMID- 8222749 TI - Management of duodenal injuries. PMID- 8222750 TI - Hand injuries. AB - A systematic approach to the injured hand has been presented that ensures no injuries are overlooked and provides the basis for a reconstructive approach. This scheme is tissue oriented, first evaluating injury to the vasculature, then to the skin, bone, joint, nerve, and tendon units. The most complex injuries can be thoroughly evaluated and properly treated with this method. PMID- 8222751 TI - Report and abstracts of the second international workshop on human chromosome 18 mapping. Doorwerth, the Netherlands, July 19-20, 1993. PMID- 8222752 TI - Regional localization of the bovine interleukin-2 (IL2) gene to chromosome 17q22- >q23 by in situ hybridization. AB - The interleukin-2 (IL2) gene was localized to the q22-->q23 bands of chromosome 17 in cattle using radioactive in situ hybridization. The localization confirms the recent provisional assignment of syntenic group U23 to this bovine chromosome. Increasing evidence for chromosomal banding and gene location homology within the Bovidae suggests that the IL2 gene most likely also maps to chromosome 17 in sheep, goats and buffaloes. PMID- 8222753 TI - Increased FISH efficiency using APC probes generated by direct incorporation of labeled nucleotides by PCR. AB - Probes of various sizes from the adenomatous polyposis coli gene (APC) were directly biotinylated by polymerase chain reaction (PCR) from genomic DNA. PCR labeling gave high efficiency in detection of fluorescence in situ hybridization (FISH) signals. Probes as small as 250 base pairs could be visualized through a fluorescence microscope without any image processing. PMID- 8222754 TI - Localization of the gene (RSN) coding for restin, a marker for Reed-Sternberg cells in Hodgkin's disease, to human chromosome band 12q24.3 and YAC cloning of the locus. AB - A novel 160-kDa intermediate filament associated protein, named restin (Reed Sternberg intermediate filament associated protein), is specifically expressed in the malignant cells of Hodgkin's disease and anaplastic large cell lymphoma (Ki-1 lymphoma). The combination of chromosomal R-banding and fluorescence in situ hybridization (FISH) with the use of two fluorescent dyes, fluorescein isothiocyanate and propidium iodide, allowed simultaneous detection of the hybridized DNA sequence and chromosomal R-banding. By this technique, the gene coding for restin (RSN) was assigned to chromosome region 12q24.31-->q24.33, while localization of the alpha-2-macroglobulin receptor (A2MR) was refined to 12q13.1-->q13.3. To further analyze the restin gene, a 500-kb YAC clone containing the gene was isolated and analyzed. A restriction map of this area is presented. PMID- 8222755 TI - Mapping of the gene coding for a paraneoplastic encephalomyelitis antigen (HuD) to human chromosome site 1p34. AB - A cDNA clone coding for a human brain protein (HuD), which is highly homologous to the Drosophila neuronal determination protein Elav and elicits antibody formation in a high proportion of patients with paraneoplastic encephalomyelitis, was used to isolate a lambda phage recombinant clone, including a large fragment of the relevant human genomic region. The fragment proved to be an efficient probe for the precise subregional mapping of the gene by molecular in situ hybridization onto extended human prometaphase chromosomes. Analysis of banded metaphases with clear hybridization signals pointed unequivocally to the localization of the HuD gene to human chromosome band 1p34. PMID- 8222756 TI - Detection of amplified sequences at 5q11-->q13 in a homogenously staining region found by fluorescent in situ hybridization in a case of B-cell non-Hodgkin's lymphoma. AB - Gene amplification is one of the molecular mechanisms possibly involved in the initiation of tumorigenesis. Homogenously staining regions and double minutes have been shown to contain amplified DNA sequences. We report here a case of B cell non-Hodgkin's lymphoma (NHL) carrying an abnormal chromosome 5 with an hsr like region at 5q11-->q13. Chromosome "painting" with a chromosome 5-specific library (pBS5) showed that the amplified material was indeed derived from chromosome 5. Further identification of the amplified DNA sequences was performed with a set of YAC probes localized to 5q12-->q14. The amplified region contained genetic loci defined by D5S6, D5S125, D5S112, and D5S39. A three-fold level of DNA amplification was visualized by Southern blotting using a DNA probe from the amplified region. The present results suggest that unknown gene(s) at 5q11-->q13 might be involved in the process of lymphomagenesis when amplification occurs. PMID- 8222757 TI - Localization of the human cholecystokinin-B/gastrin receptor gene (CCKBR) to chromosome 11p15.5-->p15.4 by fluorescence in situ hybridization. AB - A human cholecystokinin-B/gastrin receptor cDNA (CCKBR) has been recently cloned and its transcriptional product has been characterized. The cDNA probe was mapped by fluorescence in situ hybridization to chromosome 11 at bands p15.5-->p15.4. The localization of this receptor gene provides a useful marker for this region of chromosome 11 permitting the identification of diseases involving the gene and interactions with other genes. PMID- 8222758 TI - Simple sequence repeat length polymorphisms mapped to rat chromosome 11. AB - Two genes and two anonymous DNA loci were mapped to rat chromosome 11 using F2 intercross progeny of Fischer (F344/N) and Lewis (LEW/N) inbred rats. These four loci formed a single linkage group covering 21.5 cM with the following map order: somatostatin (SST)-D11N161-D11N18-cell surface protein (MOX2). These four loci were typed by PCR-based simple sequence repeat (SSR) length polymorphism detection. For each marker four to seven different alleles were detected using a panel of 13 inbred rat strains (F344/N, LEW/N, BN/SsN, BUF/N, LER/N, MR/N, MNR/N, LOU/MN, ACI/N, WBB1/N, WBB2/N, SHR/N, WKY/N). Comparative gene mapping analysis suggests syntenic conservation between rat chromosome 11 and mouse Chromosome 16. PMID- 8222759 TI - Localization of the telomeric (TTAGGG)n sequence in chicken (Gallus domesticus) chromosomes. AB - The distribution of the highly conserved eukaryotic telomeric (TTAGGG)n sequence was investigated in chicken metaphase chromosomes using the FISH technique. Besides the expected telomeric locations, interstitial as well as centromeric locations of the (TTAGGG)n repeat were observed on several macrochromosomes. The microchromosomes display three discrete patterns of labeling with this repeat. The significance of this extreme-different distribution of the telomeric related sequence in chicken chromosomes may lie in pointing to structural events that might have occurred during the process of karyotype evolution. PMID- 8222760 TI - First meiotic division abnormalities in human oocytes: mechanism of trisomy formation. AB - Trisomy is the single most frequent type of chromosome abnormality in humans and has considerable impact on many aspects of human pathology. It arises most commonly through "nondisjunction" at maternal meiosis I, but the underlying mechanism of formation remains obscure. Analysis of 100 haploid oocytes at second meiotic metaphase shows that the only type of chromosome abnormality compatible with trisomy formation after fertilisation is the presence of single chromatids in addition to, or replacing, whole chromosomes. The mechanism resulting in the presence of single chromatids is considered to be precocious division of univalents or dyads at first meiotic anaphase. PMID- 8222762 TI - Digitized and differentially shaded human chromosome ideograms for genomic applications. AB - Human chromosome ideograms that depict 850 bands, numbered in agreement with ISCN (1981) nomenclature but based on actual measurements of band sizes and differentiated by five shades of staining intensity, have been digitized using commercially available graphics software. They provide for more accurate estimates of distances within chromosome arms, DNA content of chromosome bands and correlation between cytogenetic, molecular and genetic linkage maps than the standard ISCN ideograms. PMID- 8222761 TI - Two simple procedures for releasing chromatin from routinely fixed cells for fluorescence in situ hybridization. AB - We describe two methods for releasing chromatin from routinely harvested and fixed cells. Using fluorescence in situ hybridization with combinations of probes from the HLA class II region, we show that good signals can be obtained on free chromatin fibers enabling determination of relationships between closely adjacent or overlapping probes. PMID- 8222763 TI - Fibrinolysis in patients with acute ischaemic heart disease. With particular reference to systemic effects of tissue-type plasminogen activator treatment on fibrinolysis, coagulation and complement pathways. AB - The plasminogen activator systems in the blood, the coagulation system, and the complement pathways are reviewed. The review describes the role of the vascular intima in activation of coagulation and fibrinolysis and the interrelations between the complement system and haemostatic mechanisms. Physiological activation of fibrinolysis may be triggered by and limited to fibrin because of a special affinity of plasminogen and plasminogen activators. The binding of plasminogen to fibrin is regulated by histidine-rich glycoprotein, and the primary physiological inhibitor of generated plasmin is alpha 2-antiplasmin and especially the plasminogen-binding form of this immediate plasmin inhibitor. Plasminogen activator inhibitors in the blood, that is, notably plasminogen activator inhibitor type 1 (PAI-1), bind circulating tissue-type plasminogen activator (t-PA). However, local fibrinolysis in vivo mediated by t-PA may be independent of complex formation between plasminogen activator inhibitors and t PA in the fluid phase. Circulating plasminogen activator inhibitors might regulate fibrinolysis by increasing the clearance of t-PA from the blood. The urokinase-type and factor XII-dependent fibrinolytic proactivator system can be activated following t-PA-mediated generation of plasmin, and could thus serve as an amplification system of t-PA-induced fibrinolysis. It is claimed that the as yet uncharacterized proactivator is essential for optimal generation of plasminogen activator activity by the factor XII-dependent fibrinolytic system. The normal antithrombotic condition of the vascular intima probably results from lack of tissue factor activity and the presence of significant antithrombotic components comprising, among others, antithrombin III and the protein C-protein S system. A number of pathophysiologic stimuli, notably mediators of the acute phase response such as the cytokines interleukin-1 and tumour necrosis factor alpha (cachectin), have the potential to induce the vascular endothelium to express procoagulant activity. Vascular endothelium promoting coagulant activity releases increased amounts of t-PA antigen and PAI-1 antigen into the circulation, and elevated levels in the blood of both may be regarded as a marker of a generalized procoagulant condition involving the vascular endothelium. In a prospective study in patients with unstable angina pectoris, patients in whom disease progresses and acute myocardial infarction develops, have increased amounts of t-PA antigen and PAI-1 antigen in the blood. This suggests that the procoagulant potential and atherosclerotic process of the vascular intima is more pronounced in the risk group.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8222764 TI - The influence of ethanol and liver disease on sex hormones and hepatic oestrogen receptors in women. AB - In contrast to the numerous studies of men, very few studies have been concerned with sex hormone disturbances in women with chronic alcoholic and non-alcoholic liver diseases. The aim of the study was, to evaluate the effect of ethanol and liver dysfunction on menstrual cycle, serum sex hormone concentrations and hepatic oestrogen receptors in women. In premenopausal female alcoholics ethanol consumption increase the frequency of menstrual disturbances, abortions, and miscarriages, while infertility is not frequent. Acute ethanol intoxication has only minor effects on pituitary-gonadal hormones in premenopausal women, while chronic ethanol abuse lead to reduced concentrations of sulphated steroids, and these changes may be seen before severe liver dysfunction has appeared. In women liver dysfunction lead to earlier occurrence of menopause in comparison with normal controls, while information is insufficient or lacking regarding the influence upon fertility, pregnancy outcome and sexual behavior in women. In postmenopausal women with alcoholic and non-alcoholic liver disease, the main disturbances of sex hormone metabolism consist of elevated oestrone and sex hormone binding globulin (SHBG) concentrations, while serum concentrations of steroid sulphates and 5 alpha-dihydrotestosterone (DHT) are reduced, and the degree of liver dysfunction is a major determinant for the observed disturbances. The presence of high affinity, low capacity, specific oestrogen receptors (ER) in the liver is confirmed using a ligand binding assay (DCC), specificity analyses, and sucrose gradient centrifugation. Furthermore, the sensitivity of an enzyme immunoassay has been improved enabling the quantitative measurement of hepatic ER in 102 small liver biopsies from patients with alcoholic and non-alcoholic liver diseases. The method is suitable for quantitative assessment and ER in small tissue samples, and can be applied to other tissues than the liver. Patients with chronic liver diseases have significantly lower hepatic ER concentrations, and this reduction is determined by the degree of liver dysfunction, while the degree of alcoholic hepatitis or active ethanol consumption are less important factors. The importance of this observation for hepatic physiology and pathophysiology remains to be determined. PMID- 8222765 TI - Assessment of the venous muscle pump function by ambulatory strain gauge plethysmography. Methodological and clinical aspects. PMID- 8222766 TI - Expression of NCAM and cadherins under normal and pathological conditions. PMID- 8222767 TI - Risk factors for hip fractures. A review. AB - Risk factors for hip fracture have been reviewed based on case-control and prospective follow-up studies. Falls with impact to the hip, the capacity of energy absorption in soft tissue, and bone strength are closely linked to the risk of hip fracture. Consistently documented risk factors are age, sex, race, low bone mass, low body weight, estrogen deficiency and earlier fracture. Other risk factors are tendency to fall, disability and immobilisation, low physical activity, use of psychotropic/anxiolytic/hypnotic drugs, use of corticosteroid, low calcium intake in the elderly, osteomalacia, thyreotoxicosis, cigarette smoking, chronic alcoholism, diabetes mellitus, insufficient sunlight exposure and a protective effect from thiazide diuretics. Evidence is lacking for risk factors such as heredity, nutrition and medical conditions. No information is available on the combined effects of the different risk factors. Risk estimation of hip fractures should focus on the risk of falls, the capacity of energy absorption and the bone strength. Recent studies indicate that the prevention of hip fractures is realistic, even in the elderly and definitely osteoporotic population when these fundamental risk factors are modified, which has been shown in controlled trials in nursing homes by vitamin D and calcium supplementation or by wearing external hip protectors. PMID- 8222768 TI - The effect of a community-based cardiovascular disease prevention project in a Danish municipality. AB - A community-based project for the prevention of cardiovascular diseases was undertaken in 1989 in a rural Danish municipality (Slangerup) with about 8000 inhabitants. Project goals were to draw attention to project activities and improve smoking, eating and exercise behaviours. The intervention was planned using the social learning theory, a communication-behaviour change model and community organisation principle. The strategy used for intervention involved both mass communication and active involvement of the local population in group activities. The objectives of the intervention were assessed by data obtained from representative cross-section surveys in intervention and a control area at baseline (1989) and one year later. More respondents in the intervention (82%) than control (67%) area were aware of local health projects. Ten % reported that they stopped smoking within the last year, 39% ate less fat, and 28% did more exercise, with no differences between intervention and control area. Several explanations are proposed for the limited effect of the project on behaviours. One possible explanation is that the project almost ended up being a pure mass media campaign which may increase awareness, but, as experience shows, may have limited influence on adoption of new behaviour. The Danish population around 1990 is very well informed and educated in this field due to earlier nationwide interventions. No further behavioural effects are obtainable with mass media campaigns. PMID- 8222769 TI - Cerebrospinal fluid ferritin in patients with meningitis and cerebral infarction or bleeding. AB - The diagnostic value of cerebrospinal fluid (CSF) ferritin was assessed in 30 patients with meningeal reaction (viral meningitis 10; bacterial meningitis 6; meningism 14) and in 37 patients with cerebrovascular disease (cerebral infarction 29; transient ischaemic attacks 4; cerebral haemorrhage 4). The control group comprised 13 subjects with minor neurological disorders. CSF ferritin levels were not significantly different in the various subgroups, most patients having values within the normal reference interval (2-7 micrograms/l). A clearly elevated CSF-ferritin was seen in one patient with bacterial meningitis and in one patient with intraventricular bleeding. In patients with meningeal reaction, CSF-ferritin was significantly correlated to CSF-protein. Patients with cerebrovascular disease displayed significant correlations between CSF-albumin and CSF-IgG and between CSF-ferritin, CSF-albumin and CSF-IgG. The present study indicates that measurement of CSF-ferritin is of no practical clinical relevance in the evaluation of patients with meningeal reaction and cerebrovascular disease. PMID- 8222770 TI - Cardiac performance and thyroid function. The correlation between systolic time intervals, heart rate and thyroid hormone levels. AB - In the search for a clinical correlate to thyroid function, correlations were sought between the plasma level of thyroid hormones and the cardiac function, assessed by heart rate and the systolic time intervals. The present study is the first to 1) use a curvilinear correlation and 2) do so in the entire hormone range. A third degree Chebychew's regression equation was used to enable curvilinear correlation. In a population of 10 hypothyroid, 14 euthyroid, and 25 hyperthyroid patients a statistically significant curvilinear correlation was found between the thyroid hormone levels and the systolic time intervals. The correlation coefficients showed the heart rate equally correlated to the thyroid hormone levels as the best correlated of the systolic time intervals. The correlation between the heart rate and T3 was almost linear, with a heart rate increase of 4 beats/minute with each increase in plasma T3 of 1 nmol/l. CONCLUSION: The thyroid function reflects in the cardiac function in a curvilinear regression between T3 and systolic time intervals and in an almost linear regression between T3 and heart rate. The variation, however, is too great and the slope too low, to suggest the use of the heart rate as a clinical correlate to thyroid function. PMID- 8222771 TI - The fibrinolysis and coagulation systems in ischaemic heart disease. Risk markers and their relation to metabolic dysfunction of the arterial intima. AB - This report reviews the major haemostatic deviations associated with the evolution of ischaemic heart disease (IHD) and also such deviations, which might be of importance for the evolution of the acute ischaemic heart syndrome. It is demonstrated that deviation in the t-PA/PAI-1 system indicate endothelial cell dysfunction in patients with IHD. Different factors which have the capability to induce such an endothelial cell dysfunction are proposed, i.e. hyperinsulinemia, mediators of chronic disease phase response, and thrombin. Finally, the intimate interrelation between coagulation and fibrinolysis in patients with IHD is discussed. The experimental and clinical studies reviewed provide good evidence that the endothelial cell response of t-PA and PAI-1 is coupled and that there is an intimate interrelation between generation of thrombin and production/release of t-PA and PAI-1 in IHD patients. PMID- 8222772 TI - Prevention of Pneumocystis carinii pneumonia relapse in AIDS patients. The efficacy and tolerability of low-dose sulfamethoxazole-trimethoprim. AB - The effectiveness and tolerability of Sulfamethoxazole with Trimethoprim (SMX TMP), a dose of 400mg/80mg given twice a day as secondary prophylaxis (SP) against Pneumocystis carinii pneumonia (PCP) was assessed retrospectively in 166 AIDS patients. The mean observation period was 9.7 months (range 1.0-1.4). Relapse of PCP occurred in eight patients; four episodes were histologically verified and four episodes were clinically assumed. The relapse rate after one year of prophylaxis was 5.1% (95% CI 0.0%-11.0%) using the log-rank test. Intolerance of secondary prophylaxis, defined as adverse effects necessitating cessation of SP with SMX-TMP, was reported in eight patients (5%) (95% CI 2.1% 9.3%). PMID- 8222773 TI - Topical tromantadine in the treatment of genital herpes. A double-blind placebo controlled study. AB - A randomised double-blind placebo controlled trial was performed to assess the efficacy and safety of tromantadine ointment 1% in male patients with genital herpes. 45 patients with penile genital herpes of less than three days duration were randomised to local treatment with tromantadine ointment 1% or placebo (ointment base alone) five times daily up to 12 days. No difference in time of healing could be detected between the two groups (p = 1.0). Both the objective signs (vesicles, pustules, ulceration, and glandular swelling) and the subjective symptoms (itching, burning, pain and dysuria) all diminished simultaneously in the two groups. PMID- 8222774 TI - Statistical pitfalls in the comparison between two commercial serum ferritin kits, Pharmacia and Amersham. AB - Two commercial ferritin kits, Phadebas Ferritin PRIST (kit B) and Ferritin RIA Amersham (kit A) were compared in order to "translate" ferritin values from one kit to the other. Ferritin levels in 222 sera were determined with both kits in the concentration range 5-838 micrograms/l. Regression analysis disclosed a parabolic regression between the logarithmically transformed results obtained with the two kits. Measured kit B values of 10, 12, 15, 30 and 300 micrograms/l corresponded to calculated kit A values of 15, 17, 20, 34 and 321 micrograms/l, respectively. In kit A, storage of sera for two years at -25 degrees C in combination with freeze-thawing three times produced a minor fall (p < 0.0001) in ferritin levels with a median percent decline of 16%. PMID- 8222775 TI - Pleurodesis, pH, and thoracoscopic talc poudrage. PMID- 8222776 TI - Minimizing the risk of bronchoscopy during mechanical ventilation. PMID- 8222777 TI - "You see but you do not observe". PMID- 8222778 TI - Aerodigestive manifestations of gastroesophageal reflux. What we don't yet know. PMID- 8222779 TI - Do-not-resuscitate orders in the ICU. Why are there so few? PMID- 8222780 TI - Bronchoprovocation challenge. A clinical test whose time has come. PMID- 8222781 TI - Assessment of the patient with acute asthma in the emergency department. A factor analytic study. AB - Acute asthma is a very common medical emergency. Thus, a variety of measures are currently used to assess severity; most are logical and seem reasonably effective but have never been scientifically tested in a comprehensive manner. Additionally, simple brief measures are needed for greater specificity in describing variables and in assessing different aspects of asthma. The purpose of this study was to examine the most common clinical and objective measures utilized in the assessment of acute asthma in the emergency department for determining whether these variables comprise separate or distinct dimensions. To carry out the above purpose, we used the multivariate statistical technique of factor analysis. Measures of age, duration of attack, heart rate, respiratory rate, peak expiratory flow (PEF), FVC, FEV1 percent of predicted, accessory muscle use, dyspnea, wheezing, steroid use, and early response to treatment in 194 patients with acute asthma (mean age = 32.8 +/- 11.9 years) were factor analyzed through the method of principal components. Factors extracted with this method were then rotated orthogonally through the Varimax procedure. The first factor contained the three measures of lung function (PEF, FVC, and FEV1 percent of predicted) and early response of treatment. The second factor extracted contained respiratory rate, accessory-muscle use, and dyspnea. Factor 3 contained heart rate and wheezing, and factor 4 contained age, duration of attack, and steroid use. Exploratory analysis of subgroups (by randomization, by severity of airflow obstruction, and by use of steroids) confirmed the results of the primary analysis. The coefficients of congruence obtained were above 0.90. Early response of treatment measured by the percent variation of FEV1 at 30 min over baseline value significantly correlated with a broad variety of asthma characteristics (PEF, FVC, FEV1 percent of predicted, accessory-muscle use, and duration of attack), resulting in a rather general measure, and suggesting that its measure can be a reliable assessment of acute adult asthma. Finally, early response to treatment was the most important predictor of the patient outcome (r = 0.48, p < 0.00). In summary, the results of this study support the hypothesis that most of the subjective and objective measures utilized in the assessment of patients with acute adult asthma represent separate and nonoverlapping dimensions and provided a useful summary of acute asthma. Early response to treatment in combination with measures of the four identified factors could be included in other acute adult asthma research protocols promoting greater comparable among studies. PMID- 8222782 TI - The application of an asthma severity index in patients with potentially fatal asthma. AB - The asthma severity index (ASI) is proposed as a tool to monitor the course of patients with increased risk of asthma-related morbidity and mortality. The ASI is a weighted summary of the diagnostic features of potentially fatal asthma (PFA). Fifteen patients with PFA were evaluated with ASIs calculated for the intervals 1 year prior to and 1 year following management by our allergy immunology service. In each patient, a reduction was noted in the ASI following management, which corresponded to a reduction in asthma events. The ASI will have utility in the evaluation of chronic asthma treatment and in patient education. PMID- 8222783 TI - A comparison of breath-actuated and conventional metered-dose inhaler inhalation techniques in elderly subjects. AB - BACKGROUND: Poor coordination of canister actuation and inspiration often prevents adequate metered-dose inhaler (MDI) usage by patients, perhaps especially so among the elderly. Breath-actuated inhalers (BAI) have been developed to prevent this problem. METHODS: We compared the adequacy of inhaler technique and patient preferences between MDI and BAI in a group of elderly subjects (mean age, 70.8 +/- 5.4 years). Half of the subjects were regular MDI users; half had never before used one. Two trained observers assessed the adequacy of MDI and BAI usage subjectively while performance was monitored objectively using a light source and infrared system to detect canister actuation and a spirometer to measure the inspiratory volume. If canister actuation was not followed by at least a 50 percent vital capacity, inhaler use was deemed unsuccessful. A brief teaching session preceded inhaler usage. RESULTS: By subjective assessment, BAI was used successfully more often than MDI (79 vs 60 percent, p < 0.05). By objective assessment, BAI was used successfully more often than MDI (64 vs 36 percent, p < 0.0005), although the percentage of inhalations scored adequate was lower than when assessment was subjective. Neither device was used correctly as often by those unfamiliar with MDIs as by those who were regular users. A significantly higher percentage of patients preferred BAI to MDI (71 vs 19 percent, p < 0.005), similar preferences being reported by MDI familiar and MDI unfamiliar groups. CONCLUSIONS: We conclude that (1) elderly subjects frequently handle inhalers poorly, (2) mishandling is better detected by objective than subjective monitoring, and (3) BAI is used correctly and preferred by patients more often than conventional MDIs. PMID- 8222784 TI - The lack of a role for saline solution inhalation in bronchoprovocation challenge. AB - The purpose of this study was to look prospectively at the practice of prefacing methacholine bronchoprovocation challenge (BPC) with diluent challenge using physiologic saline solution (NaCl) as the diluent. We wished to determine whether NaCl challenge added to the safety or diagnostic accuracy of BPC. We studied 108 consecutive patients undergoing methacholine BPC. We determined (1) the FEV1 response of all patients to the inhalation of NaCl (the difference between the FEV1 before NaCl and the FEV1 after NaCl), and (2) the correlation between the response to saline solution and bronchial hyperresponsiveness (BHR) measured using methacholine. Paired Student's t testing demonstrated a small but significant difference between the values for FEV1 before and after NaCl for the group as a whole (n = 108; mean change, -0.9 +/- 4 percent [+/- SD]; p = 0.023). When the mean changes in the FEV1 after NaCl for the group with increased BHR (BHR+) (n = 62; mean, -1.1 +/- 4.9 percent) and the group with no increase in BHR (BHR-) (n = 46; mean, -0.6 +/- 2.4 percent) were contrasted, there was no significant difference between the two groups (p = 0.46). Only 4 of 108 patients had a drop in FEV1 of 10 percent or more after NaCl, with the greatest drop being 16 percent. All four patients were BHR+, but none had marked BHR. For the BHR+ group, there was no correlation between response to saline solution and subsequent response to methacholine (r = 0.02). We conclude that saline solution challenge adds time and expense to BPC without increasing the safety or yield of BPC. We suggest that NaCl challenge can be omitted from the standard performance of BPC. PMID- 8222785 TI - Bronchodilator reversal of bronchospasm and symptoms incurred during methacholine bronchoprovocation challenge. Documentation of safety and time course. AB - We undertook a prospective study of bronchoprovocation challenge (BPC) to look at issues of safety and reversibility of bronchospasm and symptoms induced by BPC. Over a 14-month interval, we documented 62 consecutive cases of bronchial hyperresponsiveness. During BPC, there was a statistically significant but clinically modest increase in both cough and dyspnea. Both bronchospasm and symptoms were readily reversed with a simple protocol of inhaled albuterol using a metered-dose inhaler with a spacer. Routine protocol was effective in every case; there was never a need for individualized physician intervention. Our prospective data document the safety of BPC; we could find no reason why BPC would need to be confined to the hospital. We conclude that BPC is a valuable clinical test which merits wider dissemination and use. PMID- 8222786 TI - Bronchoconstrictive responses to inhaled ultrasonically nebulized distilled water and airway inflammation in asthma. AB - Twenty-two asthmatic patients with a range of airway hyperresponsiveness to methacholine underwent a bronchial challenge with ultrasonically nebulized distilled water (UNDW). The presence of positive responses to this stimulus was related to the extent of airway inflammation, as assessed by histochemical and immunohistochemical evaluation of bronchial biopsy specimens. Twelve patients had airflow obstruction during distilled water inhalation and they showed more severe disease than subjects with no response, as demonstrated by the higher degree of nonspecific bronchial hyperresponsiveness (p < 0.01), higher variability of peak expiratory flow rates (p < 0.01), symptom scores (p < 0.01), and daily use of bronchodilators (p < 0.01). Those patients also had increased numbers of mast cells and eosinophils (p < 0.01) and increased percentage of bronchial epithelial cells expressing endothelin 1 immunoreactivity (p < 0.01). Thus, positive responses to inhaled UNDW reflect the bronchial hyperresponsiveness consistent with moderate to severe asthma and may be due to the release of mediators with bronchoconstrictive properties from inflammatory cells or activated resident cells or both. PMID- 8222787 TI - A dose-ranging study of fluticasone propionate in adult patients with moderate asthma. International Study Group. AB - In this 4-week, multicenter, double-blind, randomized, parallel group study, the dose-effect relationship of four doses of inhaled fluticasone propionate (50, 100, 200, and 400 micrograms twice daily) was investigated and compared with beclomethasone dipropionate, 200 micrograms twice daily. A total of 672 patients with moderate asthma currently receiving 1,000 micrograms/d or less of an inhaled steroid were recruited. The study demonstrated a significant dose-related improvement in lung function with fluticasone propionate. Linear dose-related increases were observed in morning (increase per doubling dose was 4.3 L/min; 95 percent confidence interval [CI], 1.8, 6.8 L/min; p = 0.001) and evening peak expiratory flow rate (PEFR) (increase per doubling dose was 3.0 L/min; 95 percent CI, 0.5, 5.5 L/min; p = 0.017), clinic lung function (at 4 weeks, increase in percent predicted PEFR per doubling dose = 1.1 percent; 95 percent CI, 0.2, 2.1 percent; p = 0.022; increase in percent predicted FEV1 per doubling dose = 1.1 percent; 95 percent CI, 0.3, 1.9 percent; p = 0.10:increase in percent predicted FVC per doubling dose = 1.3 percent, 95 percent CI, 0.5, 2.1 percent; p = 0.001), and the percentage of symptom-free days over days 1 to 14 of treatment (increase per doubling dose = 1.9, 95 percent CI, 0.0, 3.9; p = 0.048). There was also a dose-related reduction in extra bronchodilator usage (days 1 to 14 p = 0.002; days 15 to 28 p = 0.01). In addition, there was a significant decrease in diurnal variation with increasing doses of fluticasone propionate (decrease per doubling dose = 2.0 L/min, 95 percent CI, 0.4; p = 0.024). The number of asthma exacerbations was also reduced as the dose of fluticasone propionate increased. Fluticasone propionate was well tolerated, adverse events were few, and there was a similar incidence in all groups. Furthermore, there was no evidence of any hypothalamic pituitary adrenal axis suppression. The data from the study were consistent with other clinical studies that have shown fluticasone propionate to be more potent than beclomethasone dipropionate in terms of improvement in lung function. In conclusion, this study provided evidence of a dose-related improvement in asthma control for fluticasone propionate in the dose range 100 to 800 micrograms daily, in patients with moderate asthma. PMID- 8222788 TI - Spontaneous improvement in bronchial responsiveness and its limit during preadolescence and early adolescence in children with controlled asthma. AB - To study whether improvement in bronchial responsiveness contributes to outgrowing of asthma, PC20-histamine was measured longitudinally in nine nonsteroid-dependent asthmatic children (six boys, three girls), aged 8.1 to 9.8 years (mean, 9.2 years) at the beginning of the study. The measurement was discontinued when final inhaled histamine concentration in at least two consecutive measurements did not exceed the previous highest one. Two or more doubling concentration differences in inhaled histamine is regarded as significant in our center. Mean (SD) study period was 3.6 (0.8) years, interval of the measurements was 10.6 (2.3) months. Geometric mean of PC20-histamine at the beginning was 0.39 mg/ml. PC20-histamine improved significantly in seven of nine patients during the study period, though best values were less than 8 mg/ml in all. Mean (SD) age reaching the highest final concentration was 11.0 (0.8) years. These data suggest that the improvement in bronchial responsiveness may be one of mechanisms in the outgrowing of asthma around adolescence. PMID- 8222789 TI - Occupational exposures and physician-diagnosed asthma. AB - Data from a community-based random sample of 3,606 adults 40 to 69 years of age residing in Beijing, China, were used to examine the relationship between occupational exposures to dusts and gases/fumes and physician-diagnosed asthma. The prevalence of asthma was 3.9 percent for men and 3.8 percent for women. After adjusting for sex, age, education, residential areas, indoor coal combustion, and smoking status, the attributable risks of asthma were 1.7 percent and 1.2 percent for dust and gas/fume exposure, respectively. The adjusted odds ratios of asthma for dust and gas/fume exposed groups were 1.6 (95 percent confidence interval [CI], 1.1 to 2.2) and 1.4 (95 percent CI, 0.9 to 2.1), which were independent of sex and smoking status. When subjects were classified into none, dust-only, gas/fume-only, and both-exposure groups, the estimated odds ratios of asthma were 1.3 (95 percent CI, 0.9 to 2.1) in dust-only group, 0.9 (95 percent CI, 0.5 to 1.9) in fume-only group, and 2.1 (95 percent CI, 1.2 to 3.6) in both-exposure group, suggesting a combining effect of the two agents. There was an exposure response relationship between dust and gas/fume exposures and asthma. In analysis of specific occupational agents, our findings are consistent with previously reported airway effects of organic dusts, but they also suggest that exposure to organic solvents may result in asthma, particularly when combined with dust. PMID- 8222790 TI - Work disability among adults with asthma. AB - OBJECTIVE: To estimate the incidence of work disability among adults with asthma and to evaluate a clinically based illness severity score as a predictor of such disability. DESIGN: Baseline and follow-up telephone interviews and medical record review. SETTING: University-based outpatient pulmonary specialty practice. PATIENTS: Fifty-six patients interviewed at baseline; 42 reinterviewed 2 years later. MEASUREMENTS: Work disability ascertained by interview report and defined as change in job duties, reduction in pay, or change in job or employment status attributed to asthma. Severity of asthma score derived from medical records and based on respiratory symptom frequency, asthma history, and prescribed medications. Pulmonary function by routine testing. Logistic regression analysis of the 5-year incidence of work disability on severity score and forced expiratory volume in 1 s (FEV1). RESULTS: The 5-year work disability cumulative incidence was 19 percent for change in duties, 17 percent for reduction in pay, 20 percent for change in job or work status, and 36 percent for any of these measures. The median asthma score was 10 (range, 2 to 26). The mean FEV1 as a percent predicted (FEV1 percent) was 88 +/- 25 percent. Score and FEV1 percent were statistically correlated (r = -0.6, p < 0.0001). Severity of asthma score statistically predicted each measure of work disability (p < 0.01). Addition of FEV1 percent added little additional explanatory power to the logistic regression model (maximum chi 2 = 1.3, p > 0.2). CONCLUSIONS: Work disability is common among adults with asthma. A severity of asthma score based on clinical variables is statistically correlated with lung function but appears to be a stronger predictor of disability than airflow measured at one point in time. PMID- 8222791 TI - Estimation of ventilatory reserve by stair climbing. A study in patients with chronic airflow obstruction. AB - Clinicians and surgeons have used the subjective response to the climb of "one or two flights of stairs" to assess the "reserve" of patients with chronic airflow obstruction (CAO). Very little objective data exist regarding the metabolic and ventilatory cost for any level of stair climbing in these patients. Therefore, this study was designed to evaluate the use of symptom-limited stair climbing as a simple method to estimate the peak oxygen uptake (VO2) and minute ventilation (VE) in patients with CAO. We studied 31 men with varying degrees of CAO, who climbed stairs until they stopped at their symptom-limited maximum. During this climb, timed expired gas was intermittently collected and analyzed, and oxygen saturation and heart and respiratory rates were recorded. The patients achieved 81 +/- 14 percent of their predicted maximal heart rate and 90 +/- 27 percent of their predicted maximal VE. The number of steps or flights climbed correlated linearly with peak VO2 (r = 0.72, p < 0.01) and with VE (r = 0.7, p < 0.01). Stair climbing peak VE, VO2, heart and respiratory rate correlated well with those achieved during standard leg cycle ergometry. The mean +/- SD number of flights climbed was 4.2 +/- 1.7 with most patients (87%) reaching at least 3 flights (54 steps). The group of eight patients with very severe CAO (FEV1 < 0.9 L) climbed 3.4 +/- 0.9 flights (61 +/- 16 steps). We conclude that a symptom limited maximal stair climb helps estimate peak VO2 and VE in patients with CAO. The frequently advocated test to climb one to two flights to evaluate cardiopulmonary reserve is not adequate for most patients with CAO. Symptom limited maximal stair climbing is a simple, inexpensive and readily available test that may be used to evaluate the cardiopulmonary reserve of stable patients with CAO. PMID- 8222792 TI - Bronchiectasis in patients with alpha 1-antitrypsin deficiency. A rare occurrence? AB - The chest radiographs and computed tomographic (CT) scans of seven patients with homozygous proteinase inhibitor phenotype Z (PiZZ) alpha 1-antitrypsin deficiency were reviewed. All patients except one showed severe emphysema with or without bullous change. Bronchiectasis was detected in three patients by CT but only in two patients by chest radiography. A young patient developed bronchiectasis before symptomatic emphysema. We stress that patients with PiZZ are susceptible to bronchiectasis, and the widespread use of CT should reveal its true incidence which might not be as low as generally believed. PMID- 8222793 TI - A comparison of cefpodoxime proxetil and cefaclor in the treatment of acute exacerbation of COPD in adults. AB - In this multicenter, observer-blinded study, 301 patients with signs and symptoms of acute bacterial exacerbation of COPD were randomized (2:1) to receive either cefpodoxime proxetil (200 mg, bid) or cefaclor (250 mg, tid) for 10 days. Clinical and microbiologic evaluations were performed before treatment, during therapy (study days 3 to 5), at the end of therapy (3 to 7 days posttreatment), and at long-term follow-up (4 weeks posttreatment). The most common pretreatment isolates were Haemophilus influenzae, Haemophilus parainfluenzae, and Streptococcus pneumoniae. Significantly (p < 0.001) more bacterial isolates were susceptible in vitro to cefpodoxime (233 of 256, 91 percent) than to cefaclor (215 of 255, 84 percent). There were no statistically significant differences between the two drug regimens in eradication of the initial pathogen (cefpodoxime, 116 of 128, 91 percent; cefaclor, 59 of 64, 92 percent) or end-of therapy clinical response (cure + proved; cefpodoxime, 99 of 100, 99 percent; cefaclor, 45 of 49, 92 percent) rates for evaluable patients. Both drug treatments were well-tolerated, with a similar incidence of drug-related adverse events (cefpodoxime 11 percent, cefaclor 12 percent). Cefpodoxime (bid) was as safe and effective as cefaclor (tid) in the treatment of acute exacerbation of COPD. The less frequent dosing regimen of cefpodoxime may improve patient compliance compared to those antibiotics that require three or four daily doses. PMID- 8222795 TI - Etiology of community-acquired pneumonia. Evaluation by transtracheal aspiration, blood culture, or serology. AB - In a 5-year period, 254 patients with community-acquired pneumonia were attended to. Transtracheal aspiration (TTA) could be performed on 119 patients, blood cultures were performed on 201 patients, and 74 patients underwent serologic examinations. By use of these procedures, an etiologic diagnosis was established in 93 cases. Streptococcus pneumoniae was the most common pathogen as it was found in 35 cases. Eleven of these 35 patients (31.4 percent) had pneumococcemia, and the mortality in this group was 27.3 percent. None of the patients with pneumococcal pneumonia and negative blood culture died. Haemophilus influenzae was the only isolated pathogen from transtracheal aspirated sputum in 16 cases and accounted for 17.5 percent of pneumonias in previous healthy individuals under 50 years of age. Mycoplasma pneumonia infections, Legionella pneumophila infections, and Chlamydia infections were found in ten, eight, and three cases, respectively. The overall agreement between microscopy and culture of respiratory secretions obtained by TTA was 58.8 percent, and microscopy can be a guide when choosing the initial antibiotic treatment. No statistically significant difference in the rate of isolating bacteria among patients treated with antibiotics prior to TTA and patients not previously treated with antibiotics was seen. When contraindications were respected, we found TTA to be a safe procedure. PMID- 8222794 TI - Efficacy and safety of clarithromycin compared to cefixime as outpatient treatment of lower respiratory tract infections. AB - BACKGROUND: Clarithromycin is a new acid-stable, 14-membered macrolide active against many of the organisms responsible for lower respiratory tract infections. It has been administered to over 5,000 patients worldwide and has been shown to be a safe and effective treatment for acute bacterial exacerbations of chronic bronchitis and bacterial pneumonia when given twice daily (250 to 500 mg). Cefixime is an amino-thiazolyl cephalosporin with an extended spectrum of antibacterial activity inhibiting beta-lactamase-producing respiratory pathogens. It has a long half-life, allowing once-daily administration. METHODS: This randomized, double-blind multicenter study compared clarithromycin and cefixime as treatment for patients with community-acquired lower respiratory tract infections (n = 213). Patients had bacterial pneumonia (clarithromycin, 19 percent; cefixime, 21 percent) or acute bacterial exacerbation of chronic bronchitis or asthmatic bronchitis (clarithromycin, 81 percent; cefixime, 79 percent). Patients received 500 mg of clarithromycin twice daily (n = 103) or 400 mg of cefixime once daily (n = 110) for 7 to 14 days. RESULTS: Clinical cure or improvement occurred in 86 percent of the clarithromycin-treated patients and 88 percent of the cefixime-treated patients. When only patients with identified infections with Haemophilus influenzae, Moraxella catarrhalis, or Streptococcus pneumoniae were considered, clinical success rates were 97 percent for clarithromycin and 96 percent for cefixime; the rate of bacteriologic eradication was 91 percent for clarithromycin and 90 percent for cefixime. Adverse events occurred in 29 percent of the clarithromycin-treated patients and 23 percent of the cefixime-treated patients. CONCLUSIONS: This study demonstrates that clarithromycin and cefixime are effective treatments for pneumonia and acute bacterial exacerbations of bronchitis of mild to moderate severity caused by the most common infecting organisms. PMID- 8222796 TI - A clinical comparison between Mycobacterium avium and Mycobacterium intracellulare infections. AB - Susceptibility, clinical features, and response to treatment were compared between 29 cases of Mycobacterium avium infection and 43 cases of Mycobacterium intracellulare infection detected in the Nagasaki (Japan) area and identified by a DNA probe method. In vitro susceptibility of two species to antituberculous agents was determined by a microdilution method, and M avium was more resistant to enviomycin at 25 mg/L than M intracellulare, while M intracellulare was more resistant to isoniazid at 5 mg/L and to cycloserine at 20 mg/L. No significant difference was found between infections caused by two species as to background factors, laboratory data, clinical symptoms, and chest radiographic findings at the onset of the disease. Approximately 70 percent of the patients in each group had underlying diseases; among them, pulmonary tuberculosis was the most common. Negative conversion of bacilli during the 6-month treatment was seen in 17 of 29 patients (59 percent) with M avium infection and in 21 of 43 patients (49 percent) with M intracellulare infection. Bacilli-negative conversion was slightly faster in the former than in the latter. However, these differences were statistically not significant. In conclusion, most M avium-intracellulare complex organisms are clearly identified as M avium or M intracellulare by the DNA probe method, and there was no significant difference in clinical features and response to treatment between infections caused by the two species. PMID- 8222797 TI - Early bacterial infections in lung transplant recipients. AB - Early bacterial pulmonary infections within 2 weeks after lung transplantation were studied in 29 patients undergoing surgery between December 1989 and May 1992. Suspected pulmonary infections occurred in 11 patients (38 percent). The most common bacterial organisms isolated were Klebsiella pneumoniae (45 percent; 5/11), Pseudomonas aeruginosa (36 percent; 4/11), Escherichia coli (27 percent; 3/11), Staphylococcus aureus (18 percent; 2/11), and Enterobacter cloacae (18 percent; 2/11). The mortality due to infection was 3 percent (1/29) in the early postoperative period. None of the following variables was found to be of prognostic significance: positive donor cultures, ischemic time of the graft, use of cardiopulmonary bypass, number of courses of methylprednisolone for acute rejection, duration of postoperative intubation, and type of surgical procedure. The presence of infection in the early postoperative period did not influence long-term survival. In the absence of prognostic parameters, prompt adjustment of antibiotic therapy to the results of antibiograms remains the most important therapeutic step in the management of infections in the early postoperative period after lung transplantation. PMID- 8222798 TI - Repetitively negative changing T waves at 24-h electrocardiographic monitors in patients with the chronic fatigue syndrome. Left ventricular dysfunction in a cohort. AB - This study surveys the occurrence of repetitively negative to flat T waves, alternating with normal upright T waves in 24-h electrocardiographic recordings from a subspecialty infectious diseases outpatient practice during the years 1982 to 1990. Patients with normal resting electrocardiogram in the assayed leads, but with repetitively inverted to isoelectric abnormal T waves at Holter monitors, were considered to have abnormal readings. A total of 300 patients had undergone a 24-h Holter monitor. This group included 24 individuals with chronic fatigue syndrome (CFS). This population was restricted to individuals 50 years old or younger, and the patients with CFS are compared with the patients without CFS. One of the more striking differences between the two groups was the difference in abnormal Holter readings. The patients with CFS all had abnormal Holter readings, while 22.4 percent patients without CFS had abnormal readings (p < 0.01). We further report the occurrence of mild left ventricular dysfunction in 8 of 60 patients in continuing studies of this population with CFS, younger than 50 years old, and with no risk factors for coronary artery disease. All 60 patients with CFS showed repetitively flat to inverted T waves alternating with normal T waves. Stress multiple gated acquisitions (MUGAs) (labeled erythrocytes with stannous pyrophosphate) were abnormal in eight patients with CFS. Although resting ejection fractions (EFs) were normal (mean, 60 percent), with increasing work loads (Kilopon meters [Kpms]), gross left ventricular dysfunction occurred. The fatigue of patients with CFS may be related to subtle cardiac dysfunction occurring at work loads common to ordinary living. PMID- 8222799 TI - Electrocardiographic changes in pericardial effusion. AB - OBJECTIVE: To investigate the association between ECG changes and the presence of pericardial effusion. BACKGROUND: The ECG changes associated with pericardial effusion described in textbooks are based only on small series of human cases and data from animals. These changes include low QRS voltage, electrical alternans, P wave changes, and T wave inversion. METHODS: All patients who had undergone 2 temporally separate echocardiographic and ECG examinations, with 1 echocardiographic examination indicating the presence and the other indicating the absence of pericardial effusion were identified (n = 46). These patients were age- and sex-matched to 46 patients without effusion (control subjects). Pericardial effusion was classified echocardiographically as small (n = 28), moderate (n = 13), and large (n = 5). The ECG variables were independently measured by two investigators blinded to effusion status. RESULTS: When 2 temporally separate ECGs for 46 patients were obtained in a repeated-measures fashion (1 obtained during the absence and the other during the presence of effusion; median time interval, 1.24 months), only the mean heart rate in patients with sinus rhythm (98 beats per minute increasing to 106 beats per minute) and the percentage of patients with QRS voltage of less than 0.5 mV (10 percent increasing to 22 percent) were associated with the development of effusion. A weak correlation (r = 0.296) was noted between QRS voltage and effusion size. Electrical alternans occurred only in one of the five patients with a large effusion but in no others. In addition, when the ECGs indicating effusion from the 46 patients were compared with the ECGs from their age- and sex matched control subjects, differences in heart rate (106 beats per minute vs 80 beats per minute, respectively) and small changes in QRS voltage were associated with effusion status. No ECG variable was sensitive for the detection of pericardial effusion. CONCLUSIONS: In both repeated-measures and case-control comparisons, ECG findings are two few, subtle, insensitive, and nonspecific to be useful as indicators of the presence of pericardial effusion. PMID- 8222800 TI - Antimyosin scintigraphy for detection of myocarditis. Scintigraphic follow-up. AB - To examine the value of antimyosin-indium 111 imaging in relation to endomyocardial biopsy in patients with suspected myocarditis, as well as the natural evolution of abnormal findings on the antimyosin scan, 12 patients with suspected myocarditis underwent endomyocardial biopsy and antimyosin scan. The heart-to-lung ratio (H/L) was used to quantify the antimyosin scan. All 12 patients had abnormal results on the scan (H/L, 1.7 to 2.9; mean, 2.1 +/- 0.3); 8 of the 12 patients also had a diagnostic endomyocardial biopsy. In four patients with abnormal findings on antimyosin scan and normal findings on biopsy, the H/L ratio did not differ from eight patients with abnormal findings on antimyosin scan and a diagnostic biopsy; also, the ejection fraction did not differ between the two groups. One patient died, and 8 patients had a repeat antimyosin scan within 2 months after the initial study. The H/L ratio returned to normal in two out of three patients with normal results on biopsy and in three out of five patients with a diagnostic biopsy; the ejection fraction improved by 8 percent or more in one out of three patients with a nondiagnostic biopsy and in two out of five patients with a biopsy diagnostic for myocarditis. We conclude that the antimyosin scan is more frequently diagnostic than biopsy in suspected myocarditis. Patients with abnormal results on antimyosin scan and a nondiagnostic biopsy, as well as those with abnormal results on antimyosin scan and a diagnostic biopsy, tend to return to normal results on scan within 2 months and improve their ejection fraction. PMID- 8222801 TI - Left ventricular rupture following mitral valve replacement. AB - During a 10-year period, we have encountered 6 patients (mean age, 61.2 years) with left ventricular rupture following mitral valve replacement, with an overall incidence of 1.8 percent. Four patients had early rupture, one had delayed rupture, and one had late rupture with a false aneurysm formation. Among four patients with early rupture, there were two patients with external repair by using a large ventricular patch and two patients with internal and the external repair by removing the prosthetic valve and patching both the inside and outside of the ventricle. In a patient with delayed rupture, bleeding from an epicardial hematoma was recognized along the atrioventricular groove in the intensive care unit. It was possible to control bleeding by packing the gauze, hemostatic cellulose [Surgical], and fibrin glue. Late rupture was recognized as a false aneurysm; however, there were no clinical symptoms. All patients survived the surgery, but two patients with early rupture subsequently died. One of these died of renal failure and the other died of multiple organ failure. The sites of rupture in all patients were in accordance with type 1 rupture (Treasure's classification); however, an autopsy review demonstrated the initial laceration in one case was recognized in the membranous septum 5 mm below the mitral ring and extended to the posterior atrioventricular groove. These findings suggest that the injury in the anterior mitral annulus could lead to type 1 rupture, although in the posterior mitral annulus more commonly. Since 1987, we have preserved the posterior leaflet with attached chordae when the mitral valve was fragile and myxomatous. As a result, no instances of left ventricular rupture were encountered. PMID- 8222802 TI - Complications from heparin-induced thrombocytopenia in patients undergoing cardiopulmonary bypass. AB - The purpose of this study was to evaluate retrospectively the incidence and severity of heparin-induced thrombocytopenia (HIT)-related complications in patients undergoing cardiopulmonary bypass. We reviewed the records of 1,500 consecutive patients who underwent cardiopulmonary bypass between August 1987 and December 1991 at Thomas Jefferson University Hospital. During this period of time, there were 1,155 coronary artery bypass graft operations (77 percent); 225 valve replacements and repairs, or both (15 percent); 60 combination coronary artery bypass graft or valve operations, or both (4 percent); and 60 miscellaneous procedures (4 percent). Although not all patients with postoperative complications were tested for the HIT antibody, 11 patients (0.75 percent) were diagnosed with HIT. There were 17 complications in these 5 men and 6 women including 6 cases of ischemic limbs which required amputation, 4 strokes, 2 instances of saphenous vein graft occlusion with resulting myocardial infarction, 2 cases of pulmonary emboli, 1 case of phlegmasia cerulea dolens, and 2 deaths. The complications occurred an average of 3.6 days postoperatively, with a range of occurrence of 1 to 11 days postoperatively. The mean nadir platelet count at the time of recognition was 123,000/mm3 (range 32,000 to 193,000/mm3) with 9 of 11 patients (81.8 percent) having counts greater than 100,000/mm3. There was, however, a mean percent decrease in the platelet count of 50 percent (range, 31 to 75 percent) from the time of first exposure to heparin to the time of recognition of HIT. In our patients, HIT was not related to the type, duration of treatment with or amount of heparin, or to pretreatment with aspirin. PMID- 8222803 TI - Resolution of vascular steal after pulmonary thromboendarterectomy. AB - Patients who undergo thromboendarterectomy for relief of chronic, major-vessel thromboembolic pulmonary hypertension (CT-E PH) offer a unique opportunity to evaluate potential resolution of hypertensive lesions in the small, nonelastic pulmonary arteries. Prior studies have demonstrated that, postoperatively, these patients commonly develop new perfusion scan defects. This "vascular steal" phenomenon occurs almost exclusively in lung segments which, preoperatively, were normally perfused by lung scan, were served by segmental arteries normal by pulmonary angiography, and, at surgery, were uninvolved with thrombi by direct inspection. In this study, we explored whether this intriguing "steal" phenomenon resolves over time. Twenty-nine patients who returned at 11 or more months following thromboendarterectomy were reevaluated by perfusion lung scan, repeated right heart catheterization (26 patients), and pulmonary angiography (25 patients). "Steal" of one or more lung segments occurred in 79 percent of patients in postoperative, predischarge perfusion scans. All demonstrated postoperative improvement in pulmonary hemodynamics, which persisted at follow up. Postoperative "steal" improved in 96 percent of patients and 86 percent of the "stolen" segments. The results suggest that, in CT-E PH, hypertensive lesions in the small, nonelastic pulmonary arteries are responsible for "steal," and that, with relief of pulmonary hypertension, these lesions can resolve. The study also indicates that postoperative "steal" does not connote either new thromboembolic events or a poor hemodynamic result. PMID- 8222804 TI - Postoperative complications after thoracic and major abdominal surgery in patients with and without obstructive lung disease. AB - STUDY OBJECTIVE: To determine the risk of thoracic and major abdominal surgery in patients with chronic obstructive pulmonary disease (COPD). DESIGN: Retrospective cohort study with controls. SETTING: A 692-bed teaching hospital. PATIENTS: A cohort of 26 patients with severe COPD (FEV1 < 50 percent predicted) undergoing thoracic and major abdominal surgery was matched by age and type of operation to 52 patients with mild-moderate COPD and 52 patients with no COPD. MEASUREMENTS AND RESULTS: The 26 patients with severe COPD had rates of cardiac, vascular, and minor pulmonary complications similar to patients with mild-moderate COPD and without COPD, but experienced higher rates of serious pulmonary complications (23 percent vs 10 percent vs 4 percent, p = 0.03) and death (19 percent vs 4 percent vs 2 percent, p = 0.02). All deaths and instances of ventilatory failure in the patients with severe COPD occurred in the subset undergoing coronary artery bypass surgery. Logistic regression revealed that increased age, higher American Society of Anesthesiologists class, an abnormal chest radiograph, and perioperative bronchodilator administration were associated with higher cardiac or serious pulmonary complication rates. Spirometry was not an independent predictor of postoperative complications. CONCLUSIONS: Clinical variables appear better than preoperative spirometry in predicting postoperative cardiopulmonary complications. The utility of preoperative spirometry as well as the benefits of perioperative bronchodilators in patients in stable condition remain to be determined. PMID- 8222805 TI - Video-assisted thoracic surgery. Manipulation without trocar in 112 consecutive procedures. AB - Since learning of the video-assisted thoracoscopic techniques in the treatment of thoracic diseases, we have encountered many problems and difficulties because of the limited trocar space and lack of suitable instrumentations. Since March 1992, we have found a simple and easier way to perform this procedure, that is, manipulating through the extended incisional wound without using a trocar. Endoscopic and conventional thoracic instruments are able to pass through the incisional channel freely and manipulate similarly to the technique used in open thoracotomy. All the patients had less postoperative pain, reduced operative time, and prompt return to employment. The technique is effective and is herein described. PMID- 8222806 TI - Abnormal multimeric and oligomeric composition is associated with enhanced endothelial expression of von Willebrand factor in pulmonary hypertension. AB - Abnormalities in endothelial von Willebrand factor (vWF) structure have been reported in pulmonary hypertension. These include loss of high molecular weight plasma multimers, resulting in decreased biologic activity. If endothelial processing of vWF is altered in this disorder, abnormalities in oligomeric composition may also be expected. We examined this possibility in ten adult patients with primary pulmonary hypertension. Enhanced endothelial vWF expression in these patients was indicated by increased plasma levels of vWF antigen (vWF:Ag) (214 +/- 91 vs 99 +/- 51 percent activity in controls, p < 0.001) and intense immunoperoxidase stain of pulmonary arterial endothelium for vWF (autopsy, 1 patient). Plasma from these patients also had a decreased capacity of inducing platelet aggregation in the presence of ristocetin, relative to vWF:Ag levels (57 +/- 20 percent activity). In addition to mild loss of the largest multimers, changes in oligomeric composition of plasma vWF were observed in most patients using both agarose and polyacrylamide gel electrophoresis. These included decreased concentration of dimeric (470 kDa) vWF in most patients, variable concentration of the 860-kDa fraction, and a relative decrease in subunit (223 kDa) density in subjects with elevated vWF:Ag. These findings provide additional information on the mechanisms responsible for endothelial production of dysfunctional vWF in patients with pulmonary hypertension. PMID- 8222807 TI - Stratification of patients according to prior cardiopulmonary disease and probability assessment based on the number of mismatched segmental equivalent perfusion defects. Approaches to strengthen the diagnostic value of ventilation/perfusion lung scans in acute pulmonary embolism. AB - A categorical diagnosis of "high probability" or "intermediate probability" encompasses a spectrum of diagnostic probabilities of pulmonary embolism (PE) that is not communicated to the referring physician. The diagnostic value of ventilation/perfusion lung scans, in the present investigation, was strengthened by use of a table to determine the likelihood of PE in individual patients on the basis of the observed number of mismatched segmental equivalent perfusion defects. In addition, we tested the hypothesis that stratification of patients according to the presence or absence of prior cardiopulmonary disease may enhance the ventilation/perfusion scan assessment of the probability of PE among both of these clinical categories of patients. Data were derived from the collaborative study of the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED). Ventilation/perfusion lung scans were evaluated in 378 patients with acute PE and 672 patients in whom suspected PE was excluded. Among patients with no prior cardiopulmonary disease, > or = 1.0 mismatched segmental equivalents was indicative of PE in 102 of 118 (86 percent) vs 113 of 155 (73 percent) among patients with prior cardiopulmonary disease (p < 0.02). Among patients with prior cardiopulmonary disease, > or = 2 mismatched segmental equivalents were required to indicate > or = 80 percent probability of PE. Stratification on the basis of the presence or absence of prior cardiopulmonary disease, therefore, enhanced the ability of ventilation/perfusion scan readers to assign an accurate positive predictive value and specificity to individual patients based on the observed number of mismatched segmental equivalent defects. Among patients with no prior cardiopulmonary disease, fewer mismatched segmental equivalent defects were required to indicate a high probability of PE than were required by PIOPED criteria. The findings from some of these patients, by PIOPED criteria, would have indicated intermediate probability. Some indeterminate probability readings, therefore, will be eliminated among patients stratified with no prior cardiopulmonary disease. PMID- 8222808 TI - Mismatched vascular defects. An easy alternative to mismatched segmental equivalent defects for the interpretation of ventilation/perfusion lung scans in pulmonary embolism. AB - The purpose of this investigation was to test the hypothesis that ventilation/perfusion (V/Q) lung scans in patients with suspected acute pulmonary embolism (PE) can be evaluated on the basis of the total number of mismatched vascular defects, irrespective of whether such defects are moderate or large size segmental defects. Lung scan data from the national collaborative study of the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) were assessed in 383 patients with acute PE and 681 patients in whom suspected PE was excluded. The predictive value of the cumulative number of mismatched moderate size segmental defects (irrespective of the number of mismatched large segmental defects) was nearly the same as that of mismatched large segmental defects (irrespective of the number of mismatched moderate size segmental defects). This suggests that the diagnostic value of mismatched moderate size segmental defects is the same as mismatched large segmental defects. Lung scans evaluated on the basis of the number of mismatched vascular defects (moderate and/or large segmental defects) were compared with V/Q scans evaluated on the basis of the number of mismatched segmental equivalents. The maximum likelihood estimates of the areas under the receiver operating characteristic (ROC) curves for the number of mismatched vascular defects and for mismatched segmental equivalents were similar (0.8512 vs 0.8530) (NS). Stratification according to the presence or absence of prior cardiopulmonary disease permitted a more accurate assessment of both clinical groups. Evaluation of V/Q scans by vascular defects and by segmental equivalents showed similar areas under the ROC curves. In conclusion, the number of mismatched vascular defects is as powerful for the assessment of V/Q scans as the number of mismatched segmental equivalents. The number of mismatched vascular defects, however, is easier to interpret, and permits a more objective evaluation. PMID- 8222809 TI - The addition of clinical assessment to stratification according to prior cardiopulmonary disease further optimizes the interpretation of ventilation/perfusion lung scans in pulmonary embolism. AB - The purpose of this investigation was to test the hypothesis that prior clinical assessment among patients stratified according to the presence or absence of prior cardiopulmonary disease enhances the accuracy of the predictive value of pulmonary embolism (PE) in the various categories. Diagnostic evaluation was made on the cumulative spectrum of mismatched defects, rather than a probability based on a preassigned number of mismatched segmental equivalent defects or mismatched vascular defects. Families of curves were derived that allowed an accurate assessment of the predictive value for each category of patients. The families of curves were comparable, irrespective of whether ventilation/perfusion scans were assessed on the basis of mismatched segmental equivalent defects or mismatched vascular defects, although the latter eliminated the necessity of estimating whether segmental defects were large or moderate in size. Clinical assessment was shown to prominently affect the predictive value of PE. Prior clinical assessment among patients stratified according to prior cardiopulmonary disease enhanced the accuracy of the predictive value of PE in the various groups of patients. PMID- 8222810 TI - Bronchogenic cancer in patients under 40 years old. The experience of a Latin American country. AB - Lung cancer in young patients is increasing in frequency, as documented by data from the United States, Canada, Japan, and European countries. However, to date and to our knowledge, there have not been any reports from Latin America on this topic. The published reports show that lung cancer in young patients is associated with smoking habit and family history of lung cancer. Its clinical course seems to be more aggressive than in older patients and the histologic type is less often squamous type. We describe 48 patients, aged 40 years or younger, who were diagnosed as having lung cancer in the Instituto Nacional de Cancerologia from 1980 to 1990. The patients were equally divided between men and women. Smoking was documented for only 46 percent of the cases. The histologic type most frequently diagnosed was adenocarcinoma (N = 26) followed by squamous cell carcinoma (N = 12). Almost all the cases (46 cases) were staged IV according to the TNM classification. A group of 33 patients older than 40 years (56 to 82 years) were used for comparison. The differences in sex ratio that were higher for men in the elder (m/f, 2.7:1) were family history for cancer in six patients elder; positive smoking habit in all the aged patients (100 percent) compared with only 43.7 percent in the younger group; histologic type (26 adenocarcinomas and 4 squamous in the younger compared with 14 and 12, respectively, in the elder). PMID- 8222811 TI - Survival and talc pleurodesis in metastatic pleural carcinoma, revisited. Report of 125 cases. AB - STUDY OBJECTIVE: To find out whether patients with different types of metastatic pleural carcinomas have significant differences in survival, as related to pleural fluid glucose and pH. A second objective was to evaluate the outcome of talc poudrage for pleurodesis. DESIGN: Cohort analytic prospective study; follow up of patients from thoracoscopic diagnosis and evaluation to death. SETTING: Pulmonary department at a referral medical center. PATIENTS: One hundred twenty five patients with metastatic pleural carcinoma diagnosed by thoracoscopy were evaluated prospectively. One patient was lost to follow-up. INTERVENTIONS: Thoracoscopy was performed in every patient, with talc pleurodesis performed at the end of each procedure. MEASUREMENTS AND RESULTS: On the same day as thoracoscopy or the previous day, glucose levels and pH of both the blood and pleural fluid were determined. The outcome of talc poudrage was that pleural effusions were controlled in 104 out of 119 patients (87 percent). Pleurodesis failed in 43 percent (6/14) of the patients with a pleural fluid pH lower than 7.20, as opposed to 9 percent (8/92) of failures in patients with a pH above this limit (p < 0.01). Average survival was as follows: nonsmall cell lung cancer (n = 40), 4.3 months (range, 1 to 15 months); small cell lung cancer (n = 8), 3.7 months (1 to 12 months); breast carcinoma (n = 30), 7.4 months (1 to 29 months); and ovarian carcinoma (n = 8), 9.4 months (1 to 29 months). There was a significant difference in survival between patients with breast carcinoma and patients with cancer not sensitive to chemotherapy (7.4 vs 4.7 months; p < 0.02), although the pleural tumor lesion rating was even greater in the first group. We found no significant differences between lung carcinomas and those from other origins. Patients with a low pleural fluid glucose level and low pH had significantly shorter survival than the group with high glucose and high pH levels (1.9 vs 5.7 months, respectively; p < 0.005). CONCLUSIONS: We confirmed our previous data demonstrating poor survival in patients with pleural effusions with low glucose and pH levels. The outcome of talc pleurodesis correlated to these same parameters. These results apply to all kinds of metastatic pleural carcinomas. PMID- 8222812 TI - The incidence of pleural effusion in a well-defined region. Epidemiologic study in central Bohemia. AB - Pleural effusion may complicate various diseases. To facilitate the differential diagnosis of pleural effusion, the authors conducted an epidemiologic study of incidence and etiology of different types of pleural effusions in a well-defined region in central Bohemia. During a 1-year period, the authors worked with the general practitioners and other physicians from the local hospital to identify patients with pleural effusion either while the patient was alive or at autopsy. During this period, 142 individuals (93 living and 49 at autopsy) with pleural effusion were identified among a population of 44,000 (incidence = 0.32 percent). The most common etiologies of the effusions were congestive heart failure (65, 46 percent), malignant effusions (31, 22 percent), parapneumonic effusions (24, 17 percent), and pulmonary emboli (8, 5.6 percent). Other etiologies for the effusions included pulmonary hemothorax (6), intra-abdominal processes (4), uremia (2), myxedema (1), and rheumatoid pleuritis (1). We conclude that pleural effusions are relatively common and if these figures are extrapolated to the United States and Czechoslovakia respectively, one could expect at least 800,000 cases in the United States and 48,000 in Czechoslovakia annually. Over 90 percent of the pleural effusions will be due to congestive heart failure, malignancy, pneumonia, or pulmonary emboli. PMID- 8222813 TI - Improved pulmonary function and exercise tolerance with inspiratory muscle conditioning in children with cystic fibrosis. AB - This study documented the effect of inspiratory muscle conditioning in children with cystic fibrosis. Subjects, ages 7 to 14 years, were divided into two groups. The experimental group (n = 10) trained at a high pressure load (> or = 29 cm H2O) and the control group (n = 10) trained at a minimal pressure load (< or = 15 cm H2O), using a threshold loading device. Subjects trained 30 min a day for 10 weeks. Pulmonary function, inspiratory muscle strength, and exercise tolerance were measured at the beginning and end of the training period. Pulmonary function was measured by body plethysmography. Inspiratory muscle strength was determined by standard measures of maximal inspiratory pressure against an occluded airway. Exercise tolerance was measured by the length of time subjects could walk on a treadmill. Findings indicated that the experimental group showed significant increases in inspiratory muscle strength, vital capacity, total lung capacity, and exercise tolerance in comparison to the control group. PMID- 8222814 TI - Elevated pyrogenic cytokines in heatstroke. AB - STUDY OBJECTIVES: Heatstroke, characterized by hyperthermia and neurologic abnormalities, can cause shock, adult respiratory distress syndrome, and multiorgan failure culminating in death. The mediation of metabolic changes and tissue damage is not fully understood. Recent evidence suggests the involvement of endotoxin, tumor necrosis factor alpha (TNF-alpha), and interleukin 1 alpha (IL-1 alpha) and we hypothesized that other pyrogenic cytokines may be implicated. DESIGN: Prospective analysis. SETTING: Heatstroke Center in Makkah (Mecca), Saudi Arabia. MEASUREMENTS AND RESULTS: We measured plasma IL-1 beta, IL 6, and interferon gamma (INF-gamma) concentrations by enzyme-linked immunosorbent assay in 28 heatstroke patients at the time of hospital admission (precooling) and after complete cooling (postcooling), and in 10 normal control subjects. We measured C-reactive protein (CRP) as a marker of acute phase response and calculated severity of illness using the simplified acute physiology score. Twenty-five male and 3 female subjects had mean (+/- SEM) rectal temperature of 41.2 +/- 0.2 degrees C. IL-6, IL-1 beta, and INF-gamma concentrations were elevated in 100 percent, 39 percent, and 50 percent of patients to (mean +/- SEM) 220 +/- 44 pg/ml, 42 +/- 14 pg/ml, and 1,180 +/- 879 pg/ml, respectively (normal control values: < 3.5 pg/ml, < 4.5 pg/ml, < 20 pg/ml). The CRP value was elevated in 72 percent of patients to 152 +/- 40 mg/L (control value: 0 to 17 mg/L). The IL-6 concentrations correlated with severity of illness (r = 0.516, p = 0.03); two patients with the highest concentrations died. There was no significant correlation between circulating levels of IL-6, IL-1 beta, INF-gamma, and temperature, or between IL-6, IL-1 beta, and CRP. Postcooling, IL-6, and IL-1 beta were still above normal control values; INF-gamma could be detected in one patient only. CONCLUSION: Our findings of elevated circulating IL-6, IL-1 beta, and INF-gamma in the presence of acute phase response, and correlation with severity of illness, suggest that these cytokines have a role in the pathogenesis of heatstroke, which could lead to new therapeutic strategies. PMID- 8222815 TI - Mechanism of sleep-induced periodic breathing in convalescing stroke patients and healthy elderly subjects. AB - Cerebral vascular ischemic strokes are known to precipitate Cheyne-Stokes periodic breathing. Interestingly, Cheyne-Stokes-like breathing during sleep may be associated with obstructive sleep apnea (OSA) in some individuals. Therefore, it was reasoned that stroke patients with periodic breathing in sleep would be susceptible to OSA. Because oscillations in upper airway resistance can occur as a component of sleep-induced periodic breathing, we hypothesized that stroke patients with sleep-induced periodic breathing would have oscillations in upper airway resistance. These oscillations in resistance would be expected to contribute to OSA. We studied stroke patients with sleep-induced periodic breathing and control subjects to evaluate the relationship between upper airway resistance and ventilation in periodic breathing in sleep. Ventilation and upper airway resistance were measured in presleep wakefulness and in stage 2 NREM sleep. Mean tidal volume, minute ventilation, respiratory cycle timing variables, and upper airway resistance were not different between stroke and control subjects, either awake or asleep. Upper airway resistance increased and ventilation volume decreased from wakefulness to sleep in both groups. In an equivalent number of subjects from each group, reciprocal patterned oscillations in tidal volume and upper airway resistance were present at a 5 to 12.5 breath frequency during sleep. As upper airway resistance increased, tidal volume decreased. Stroke patients had wider fluctuations in upper airway resistance than control subjects, likely contributing to the higher number of sleep-disordered breathing events observed in the stroke patients. PMID- 8222816 TI - Chronic cough due to gastroesophageal reflux. Clinical, diagnostic, and pathogenetic aspects. AB - BACKGROUND: Gastroesophageal reflux (GER) is a common cause of chronic cough. Moreover, chronic cough can be the sole presenting manifestation of GER disease (GERD). It has been suggested recently that GER most often causes chronic cough by stimulating the distal esophagus. To gain further diagnostic and pathophysiologic knowledge, we prospectively evaluated a group of patients with chronic cough likely to be due to GER with extensive gastrointestinal and respiratory studies and then observed their response to antireflux therapy. METHODS: We prospectively characterized 12 subjects whose chronic cough was likely to be due to GER by chest radiographs, barium esophagography, 24-h esophageal pH monitoring (EPM) with probes in the distal and proximal esophagus, esophagoscopy, and bronchoscopy. Then, prior to instituting antireflux therapy, we objectively counted coughs during the distal esophageal infusion of 0.1 N HCl or 0.9 percent saline solution administered in a randomized, double-blind, standardized fashion (ie, Bernstein acid-perfusion test). RESULTS: Gastroesophageal reflux was determined to cause cough in all subjects based on disappearance of cough with antireflux therapy. It was clinically "silent" in 75 percent. The EPM was the test most frequently abnormal (sensitivity, 92 percent). Distal esophageal data revealed that 10 of 12 subjects had GER-induced coughs (12 +/- 12) while only 7 of 12 had an abnormal esophageal pH conventional parameter (eg, percent time pH < 4). Compared with the distal esophagus, GER to the proximal esophagus occurred (p = 0.017) and induced cough (p = 0.004) less often. Compared with baseline (9.3 +/- 17.6), there were no differences in coughs induced by the infusion of saline solution (9.2 +/- 15.9) or acid (15.1 +/- 26.7); the number of coughs induced by acid was negatively correlated with distal esophageal acid-GER events during EPM (r = -0.64, p = 0.01). Neither bronchoscopy nor chest radiographs were consistent with aspiration. CONCLUSIONS: There is a clinical profile that prospectively predicts which patients have chronic cough due to GER. The cough was most likely due to stimulation of the distal esophagus, not aspiration. Intraesophageal acid is unlikely to be the sole mediator in gastric juice causing the cough. While EPM is the single most helpful diagnostic test, conventionally utilized diagnostic indices of GERD can be misleadingly normal; observing GER-induced coughs is more frequently helpful. PMID- 8222817 TI - Measurement of respiratory acoustical signals. Comparison of sensors. AB - We assessed the performance of three air-coupled and four contact sensors under standardized conditions of lung sound recording. Recordings were obtained from three of the investigators at the best site on the posterior lower chest as determined by auscultation. Lung sounds were band-pass filtered between 100 and 2,000 Hz and sampled simultaneously with calibrated airflow at a rate of 10 kHz. Fourier techniques were used for power spectral analysis. Average spectra for inspiratory sounds at flows of 2 +/- 0.5 L/s were referenced against background noise at zero flow. Air-coupled and contact sensors had comparable maximum signal to-noise ratios and gave similar values for most spectral parameters. Unexpectedly, less sensitivity (lower signal-to-noise ratio) at high frequencies was observed in the air-coupled devices. Sensor performance needs to be characterized in studies of lung sounds. We suggest that lung sound spectra should be averaged at known airflows over several breaths and that all measurements should be reported relative to sounds recorded at zero flow. PMID- 8222818 TI - Heterotopic ossification complicating critical illness. AB - Heterotopic ossification (HO), deposition of para-articular ectopic bone, is associated with musculoskeletal trauma and certain congenital and metabolic disorders. Additionally, HO may follow paralysis from diverse traumatic and nontraumatic neurologic insults. We describe three cases of HO associated with catastrophic nontraumatic respiratory illness requiring prolonged chemical paralysis and cardiorespiratory support. PMID- 8222819 TI - Temporal hemodynamic and oxygen transport patterns in medical patients. Septic shock. AB - STUDY DESIGN: Because of the gradual insidious transitions between localized infection, generalized infection, and septic shock, it is difficult to compare data of patients in various stages and to differentiate primary from secondary and tertiary events. The aim of the present study was to describe the sequential pattern of hemodynamic and oxygen transport patterns of survivors and nonsurvivors of septic shock in order to evaluate possible physiologic mechanisms and to provide a template to relate the sequence of physiologic events to biochemical mediators. PROCEDURE: A previously described defined protocol was used prospectively to study the sequence of physiologic events using specific criteria to define stages as: (a) early period, the first recorded change in cardiac output; (b) middle period, time of maximal metabolic activity defined as the highest recorded oxygen consumption (VO2); and (c) late period, the time of death or recovery. In addition, three time lines were defined as the first time mean arterial pressure fell below 70 mm Hg, the first time temperature rose above 38 degrees C, and the earliest fall in VO2. Physiologic data were aligned in actual time before or after the time these criteria were met. Invasive hemodynamic and oxygen transport variables were measured with systemic and pulmonary artery catheters; intravascular pressures, arterial and mixed venous gas levels, cardiac output, and derived calculations were made at frequent intervals and keyed to the time of the cardiac output; each set of measurements in turn was keyed to the aforementioned time periods to describe the early, middle, and late periods. RESULTS: Beginning with increased cardiac index and oxygen delivery (DO2) as the early physiologic changes, there were progressive increases in cardiac index, DO2, and VO2 throughout the early and middle periods. They were maintained above the normal range in the late stage of survivors, but fell in the last 16 h in nonsurvivors. These values were greater in survivors than in nonsurvivors throughout. There were early transient reductions in VO2 that preceded the rise in temperature and the fall in mean arterial pressure (MAP). Although 84 percent of the septic patients were hyperdynamic, there were transient hypodynamic episodes defined as cardiac index of less than 2.5 L/min.m2 in approximately 10 percent of the measurements. There were also transient preterminal hypermetabolic periods in about 8 percent of the nonsurvivors. CONCLUSION: We conclude that increased cardiac index and DO2 represent compensations for circulatory inadequacies that limit body metabolism as reflected by VO2. Cardiac index, DO2, and VO2 values of survivors were higher than those of nonsurvivors and normal values. Therapy directed toward increasing cardiac index to supranormal values empirically determined by survivors has been reported to improve outcome. Additional studies to describe temporal relationships of biochemical mediators of these physiologic patterns are needed. PMID- 8222820 TI - A comparative study of oral endotracheal tube securing methods. AB - A prospective study was done to compare four different methods of securing oral endotracheal tubes: adhesive tape (A), Twill tape (T), Twill tape with FlexBlue bite block (TFXB), and Velcro tie with FlexBlue (VFXB), used on sequential days. Thirty-six patients were enrolled for 136 patient-days and 18 had complete 4-day cycles. The methods were evaluated twice daily by nurses, respiratory therapist, and patient, on a five-point Likert scale with regard to oral hygiene, patient comfort, nurse satisfaction, and ease of use. Tube movement relative to the incisor teeth was measured at end of shift, the use of a bite block was noted, and near extubations were documented. Analysis of variance and Student's t test with Bonferroni correction were performed. Adhesive tape had 33 patient-days, 0.4 +/- 0.7 cm movement, 3.4 +/- 0.9 oral hygiene, 4.0 +/- 0.8 comfort, 4.2 +/- 0.6 nurse satisfaction, and 4.2 +/- 0.7 ease of use. Twill tape had 34 patient-days, 0.7 +/- 1.1 cm movement, 3.0 +/- 0.7 oral hygiene, 3.1 +/- 0.9 comfort, 2.8 +/- 1.0 nurse satisfaction, and 3.3 +/- 1.1 ease of use. Twill tape with FlexBlue bite block had 35 patient-days, 1.3 +/- 2.0 cm movement, 2.5 +/- 1.2 oral hygiene, 1.9 +/- 1.1 comfort, 1.9 +/- 1.0 nurse satisfaction, and 2.1 +/- 1.1 ease of use. Velcro tie with FlexBlue had 34 patient-days, 0.8 +/- 1.0 cm movement, 1.9 +/- 1.2 oral hygiene, 1.5 +/- 1.4 comfort, 1.6 +/- 1.0 nurse satisfaction, and 1.8 +/- 1.1 ease of use. Statistical analysis showed no difference between the groups for tube movement. Method A was statistically superior to VFXB and TFXB on all other parameters, and T on all except oral hygiene. Twill tape was superior to VFXB on all, and TFXB on patient comfort, nurse satisfaction, and ease of use. There was no significant difference between TFXB and VFXB on any measured parameter. Adhesive tape and T required an oral airway on only 14 days compared with 69 days of FlexBlue use. Extubation on 2 and near extubation occurred on 18 occasions with FlexBlue use and only once with T and accounted for most decisions to change securing method. We cannot recommend the use of the FlexBlue system for securing oral endotracheal tubes. PMID- 8222821 TI - Cardiopulmonary effects of bronchoalveolar lavage in critically ill patients. AB - Bronchoalveolar lavage (BAL) has been proposed as a useful procedure for bacteriologic diagnosis of lower respiratory tract infection in mechanically ventilated patients. To determine the cardiopulmonary effects of this procedure and to identify the patients at risk of poor tolerance, 30 critically ill ventilated patients suspected of having pneumonia were studied. Hemodynamic and gas exchange parameters were continuously recorded using an arterial catheter, a Swan-Ganz catheter with SvO2 display, and a pulse oximeter. In addition to the basal sedation required by these patients, midazolam, 0.1 mg/kg intravenously, was administered 5 min prior to bronchoscopy. A moderate increase (10 percent from basal values) in heart rate, mean arterial pressure, and cardiac index was recorded at each measurement during the procedure. A marked decrease in PaO2 was observed during bronchoscopy associated with an increase in oxygen consumption. Maximal changes in SaO2 and SvO2 were recorded at the end of BAL. Two hours after the end of BAL, PaO2 values were still 20 percent lower than pre-BAL values in 40 percent of the patients. We conclude that BAL can be performed safely in most critically ill ventilated patients who have stable hemodynamic and ventilatory parameters. However, none of the recorded parameters allows identification of the patients at risk of poor tolerance of the procedure. PMID- 8222823 TI - Mechanical insufflation-exsufflation. Comparison of peak expiratory flows with manually assisted and unassisted coughing techniques. AB - Pulmonary complications are major causes of morbidity and mortality for patients with severe expiratory muscle weakness. The purpose of this study was to compare peak cough expiratory flows (PCEFs) during unassisted and assisted coughing and review the long-term use of mechanical insufflation-exsufflation (MI-E) for 46 neuromuscular ventilator users. These individuals used noninvasive methods of ventilatory support for a mean of 21.1 h/d for 17.3 +/- 15.5 years. They relied on manually assisted coughing and/or MI-E during periods of productive airway secretion. They reported a mean of 0.7 +/- 1.2 cases of pneumonia and other serious pulmonary complications and 2.8 +/- 5.6 hospitalizations during the 16.4 year period and no complications of MI-E. A sample of 21 of these patients with a mean forced vital capacity of 490 +/- 370 ml had a mean maximum insufflation capacity (MIC) achieved by a combination of air stacking of ventilator insufflations and glossopharyngeal breathing of 1,670 +/- 540 ml. The PCEFs for this sample were: following an unassisted inspiration, 1.81 +/- 1.03 L/s; following a MIC maneuver, 3.37 +/- 1.07 L/s; with manual assistance by abdominal compression following a MIC maneuver, 4.27 +/- 1.29 L/s; and with MI-E, 7.47 +/- 1.02 L/s. Each PCEF was significantly greater than the preceding, respectively (p < 0.01). We conclude that manually assisted coughing and MI-E are effective and safe methods for facilitating airway secretion clearance for neuromuscular ventilator users who would otherwise be managed by endotracheal suctioning. Severely decreased MIC, but not necessarily vital capacity, is an indication for tracheostomy. PMID- 8222822 TI - Effects of consecutive protected specimen brushing and bronchoalveolar lavage on gas exchange and hemodynamics in ventilated patients. AB - OBJECTIVE: To assess cardiovascular effects and the oxygenation status of mechanically ventilated patients undergoing protected specimen brushing (PSB) and bronchoalveolar lavage (BAL) under fiberoptic bronchoscopy (FOB). DESIGN: A prospective study. SETTING: Polyvalent intensive care unit in a university hospital. PATIENTS: Twelve consecutive, critically ill, intubated, and mechanically ventilated patients with hemodynamic failure requiring invasive monitoring with an indwelling radial artery catheter and indwelling Swan-Ganz catheter were included in the study. INTERVENTIONS: Hemodynamic measurements, arterial and mixed-venous blood gas analyses, and arterial blood lactate analysis were performed before and at the end of a 10-min period of mechanical ventilation with a fractional concentration of oxygen in the inspired gas (FIO2) of 1.0. The same measurements and blood samplings were repeated at the end of the PSB procedure, at the end of the BAL procedure, and 1 h after the end of the BAL. During the study period the ECG, arterial oxygen saturation (SaO2), and mixed venous oxygen saturation (SvO2) were continuously monitored. MAIN RESULTS: A moderate increase in both mean arterial pressure and mean pulmonary arterial pressure was observed during the FOB procedure (p < 0.05). One hour after the end of BAL, the PaO2 decreased when compared with values recorded at the beginning of the procedure with the same FIO2 (p < 0.05). An increase in intrapulmonary shunt was observed at the end of BAL (p < 0.01). A moderate increase in PaCO2 was also observed after PSB (p < 0.05) and after BAL (p < 0.01). Monitoring of SaO2 permitted us to observe a significant and sustained decrease after the end of the FOB procedure from 10 to 60 min. The decrease in SvO2 was less pronounced but reached statistical significance. CONCLUSIONS: We conclude that PSB and BAL under FOB are well tolerated in critically ill, mechanically ventilated patients with hemodynamic disturbances requiring inotropic or vasopressor agents (or both); however, a modest impairment in arterial oxygenation was observed after the end of the FOB procedure. PMID- 8222824 TI - Spontaneous bilateral pneumothorax in a patient with mediastinal enlargement. PMID- 8222825 TI - What role for inhaled steroids in chronic asthma? PMID- 8222826 TI - Endotoxin in human disease. Part 1: Biochemistry, assay, and possible role in diverse disease states. PMID- 8222827 TI - Early inflammatory response of minocycline and tetracycline on the rabbit pleura. AB - The histopathologic findings were compared from 20 mg/kg intrapleural tetracycline hydrochloride (TCN) and three doses of intrapleural minocycline hydrochloride (5, 10, and 20 mg/kg) (MCN) in New Zealand white rabbits. Both TCN and MCN produced an early neutrophilic predominant pleural effusion that became mononuclear over 48 h. There was no difference in pleural fluid accumulation, number of adhesions, or histologically measured visceral and parietal pleural thickness between TCN and MCN (all p = ns). The TCN, 20 mg/kg, produced more visceral pleural plaque than MCN, 5 mg/kg (p < 0.05). Increasing MCN doses resulted in greater pleural fluid neutrophil accumulation. With higher dose MCN, greater mesothelial cell desquamation and fibroblast proliferation was evident compared to the 5 mg/kg dose. The MCN and TCN produce similar histopathologic condition in the rabbit pleura which suggests that MCN should cause a similar clinical response in humans. PMID- 8222828 TI - Smoking restrictions at Boston-area hospitals, 1990-1992. A serial survey. AB - Boston-area hospitals were initially surveyed in 1990 and again in 1992 regarding their policies on smoking. In 1990, all 40 facilities participating had policies restricting in-hospital smoking, 3 had banned smoking entirely, and only 1 hospital still permitted tobacco sales on its grounds. By December 1991, 18 of the hospitals had banned smoking, and no facility sold tobacco. As of February 1992, 29 had banned smoking, and 37 were offering ongoing smoking cessation programs to their employees. A new Joint Commission on Accreditation of Healthcare Organizations standard requiring hospitals to be smoke-free by the end of 1993 seemed to have a strong effect on the implementation of smoking bans. PMID- 8222829 TI - Use of do-not-resuscitate orders in an intensive care setting. AB - STUDY OBJECTIVES: Do-not-resuscitate (DNR) orders have been espoused for the enhancement of patient autonomy, avoidance of futile medical intervention, and cost containment. Outcomes of cardiopulmonary resuscitation (CPR) in the intensive care setting have been dismal, with few patients surviving to discharge. This study compares patients who died in medical and surgical ICUs in a DNR status with those who died after attempted CPR. DESIGN: Retrospective chart review of all patients who died in the medical and surgical ICU in a 2-year period. MEASUREMENTS AND RESULTS: A total of 195 cases were reviewed during the specified time period; 108 patients had undergone attempted resuscitation, and 87 patients died in a DNR status. There were no significant differences when preadmission disability, source of admission, location (medical ICU vs surgical ICU), chronic medical conditions, acute diagnosis, sex, and weight were considered. Patients who were designated "DNR" were significantly older than patients who underwent CPR (mean age, 65.7 years vs 58.9 years; p = 0.005). The DNR-designated patients were in general more severely ill as measured with the APACHE II system (mean score, 23.5 vs 20.7; p = 0.004), which was accounted for primarily by greater alterations in level of consciousness as measured with the Glascow Coma scale (mean score, 10.0 vs 12.1; p = 0.001). CONCLUSIONS: Among patients dying in the medical and surgical ICUs in the authors' institution, only age and level of consciousness discriminated patients who died in a DNR status from those who died after attempted CPR. PMID- 8222830 TI - Persistent bilateral effusions and fever in a 48-year-old man. PMID- 8222831 TI - Ultrasonography and color Doppler imaging of metastatic pulmonary choriocarcinoma. AB - Chest ultrasound and color Doppler flow imaging were used to assess the pulmonary nodule of a 24-year-old pregnant woman with dyspnea and hemoptysis. The ultrasonogram showed multiple hypoechoic subpleural nodules. Color Doppler imaging of the nodules revealed characteristics of high vascularity and high blood flow. A percutaneous needle aspiration under ultrasonic guidance confirmed the diagnosis of choriocarcinoma. PMID- 8222832 TI - Synchronous lung cancer presenting with small cell carcinoma and adenocarcinoma. AB - We describe a case of synchronous primary lung cancer presenting with small cell carcinoma and adenocarcinoma, in which expression of the cell surface antigens and also tumor markers were evaluated immunohistologically. A review of the literature concerning synchronous or metachronous primary lung cancers is also presented. PMID- 8222833 TI - Thrombus of the ascending aorta as a source of cerebral embolism. AB - A 74-year-old woman presented with a stroke. Transesophageal echocardiography showed evidence of a thrombus attached to the wall of the ascending aorta that was not detected by the transthoracic approach. The thrombus was no longer present after treatment with heparin with no recurrent embolic event. To our knowledge, this is the first report of a thrombus attached to the ascending aorta that was no longer present after treatment with heparin, was detected by transesophageal echocardiography, and was thought to be the source of cerebral embolic event. PMID- 8222834 TI - An atypical carcinoid tumor of the lung with mutations in the p53 gene and the retinoblastoma gene. AB - Molecular analysis of a metastatic lesion of an atypical carcinoid tumor of the lung obtained from a 77-year-old man at autopsy revealed a point mutation in the p53 gene and a deletion in the retinoblastoma (Rb) mRNA. This case suggests that both these antioncogenes may be involved in the progression of atypical carcinoid tumor. PMID- 8222835 TI - Fatal Haemophilus influenzae septicemia following bronchoscopy in a splenectomized patient. AB - We describe a 46-year-old splenectomized patient who died of Haemophilus influenzae septicemia 16 h following bronchoscopy. Although rare, postsplenectomy overwhelming sepsis is always a danger in splenectomized patients undergoing invasive procedures. Chemoprophylaxis should be considered in asplenic patients peribronchoscopy. PMID- 8222836 TI - Cardiogenic shock due to huge right atrial thrombus. AB - An unusual case is reported of right atrial thrombus associated with cardiogenic shock. The role of two-dimensional echocardiography in detection of blood clots is highlighted. PMID- 8222837 TI - Severe pneumococcal pneumonia complicated by massive pulmonary gangrene. AB - Massive pulmonary gangrene is a rare complication of pneumonia, particularly in the postantibiotic era. We report two cases of community-acquired Streptococcus pneumoniae pneumonia in young patients with a background of heavy alcohol abuse, but no other preexisting disease, which failed to respond to appropriate antibiotic therapy and intensive care. In both, there was extensive unilateral involvement, with initial dense consolidation followed by cavitation, but the previously reported classic later radiologic feature of coalescence into a large cavity with free-floating slough was not seen. Owing to ongoing sepsis with the development of multiple organ failure and the obvious failure of appropriate medical therapy, both patients underwent pneumonectomy with a successful outcome. These cases serve to emphasize the role of surgery in the management of massive pulmonary gangrene. PMID- 8222838 TI - Ischemically mediated sustained monomorphic ventricular tachycardia. Resolution with anti-ischemic therapy. AB - We describe a patient with ischemically mediated sustained monomorphic ventricular tachycardia occurring at rest and resistant to treatment with intravenously administered procainamide. Percutaneous transluminal coronary angioplasty was initially successful, but rest angina and ventricular tachycardia, resistant to procainamide therapy, recurred 2 weeks later and responded to aggressive anti-ischemic medical therapy. We suggest that anti ischemic medication may be of benefit in patients with malignant ventricular arrhythmias precipitated by spontaneous myocardial ischemia. PMID- 8222839 TI - Unmasking accessory pathway conduction with adenosine-induced atrioventricular nodal block after radiofrequency catheter ablation. AB - Radiofrequency catheter ablation is very effective in eliminating conduction over accessory pathways in patients with Wolff-Parkinson-White syndrome. However, accessory pathway conduction recurs in approximately 5 to 9 percent of patients in the weeks to months following ablation. We describe two cases in which intravenous adenosine revealed persistent accessory pathway conduction after apparently successful ablation, thus providing an indication for the delivery of further ablative therapy. Adenosine may improve the long-term efficacy of radiofrequency catheter ablation of accessory pathways by manifesting latent accessory pathway conduction. PMID- 8222840 TI - Double-orifice mitral valve with three papillary muscles. AB - A case of double-orifice mitral valve in a 16-year-old Japanese boy is presented. Two-dimensional echocardiography, including transesophageal approach, showed two approximately equal orifices with three papillary muscles. Color Doppler echocardiography showed a mild mitral regurgitation from both orifices. No other cardiac anomaly was associated. The patient's left ventricular function is presently normal. PMID- 8222841 TI - Amrinone therapy for severe pulmonary hypertension and biventricular failure after complicated valvular heart surgery. AB - We report two cases in which amrinone was used effectively, in addition to the conventional sympathomimetic drug, for the emergence from cardiopulmonary bypass following complicated valvular heart surgery in patients who had severe pulmonary hypertension and biventricular failure. Amrinone was used in combination with isoproterenol in one and dopamine in the other case. The clinical changes were brought about by a 21.5 percent and 53.5 percent decrease in pulmonary blood pressure and pulmonary vascular resistance, respectively. Concomitantly, the mean systemic blood pressure was increased by 50 percent, whereas heart rate decreased by 17.5 percent. This report demonstrates that amrinone can be life-saving in patients with biventricular failure and severe pulmonary hypertension not responding to conventional beta-adrenergic and vasodilator drug therapy. PMID- 8222842 TI - Pneumonitis complicating low-dose methotrexate therapy for rheumatoid arthritis. Discrepancies between lung biopsy and bronchoalveolar lavage findings. AB - Two very similar cases of drug-induced pneumonitis complicating treatment of rheumatoid arthritis with low-dose methotrexate are presented. Diagnosis was suggested by clinical history and findings, but the bronchoalveolar lavage showed a high percentage of neutrophils, an unusual feature in methotrexate-induced pneumonitis. Transbronchial lung biopsies (TBB) confirmed the diagnosis by showing interstitial lymphocytic infiltrate with microgranulomas. Although histologic findings are not strictly pathognomonic, when a differential diagnosis has to be made with infectious and rheumatoid lung disease, TBB appears to be of great promise. PMID- 8222843 TI - Extramedullary hematopoietic tumors of the posterior mediastinum related to asymptomatic refractory anemia. AB - Two asymptomatic paravertebral thoracic masses occurred in a 65-year-old patient with isolated macrocytosis. The largest one measured 8 cm and was surgically resected with a presumptive diagnosis of schwannoma. This thoracic mass was hemorrhagic, encapsulated, and composed of fat and hematopoietic tissue. While extramedullary hematopoietic tumors usually occur in patients with severe chronic hemolytic anemia, our report suggests that such lesions must be considered in the differential diagnosis of posterior mediastinal mass in patients without clinical evident anemia. PMID- 8222844 TI - Increased blood pressure during inverse ratio ventilation in two patients with adult respiratory distress syndrome. AB - Inverse ratio ventilation (IRV) is increasingly used in the supportive treatment of patients with hypoxemic respiratory failure. A recent study suggests that IRV reduces cardiac output with minimal effect on mean arterial pressure. We report two cases in which IRV led to reproducible increases in mean arterial pressure. Concomitant hemodynamic measurements suggest that these responses occurred as a result of increased vascular resistance. PMID- 8222845 TI - Cardiac operation without hypothermia for the patient with cold agglutinin. AB - Recently, a new technique for myocardial protection that does not rely on hypothermia has been reported. In this method, the heart is continuously perfused with normothermic hyperkalemic blood cardioplegia during the cross-clamp period. Cardiac arrest is achieved and maintained using high levels of potassium. Hypothermia is not part of this technique; thus, the danger of hypothermia can be avoided in the patient with cold agglutinin disease without compromising myocardial protection. This communication reports our experience using retrograde continuous normothermic blood cardioplegia in one patient with potent cold agglutinins and severe coronary artery occlusive disease. This patient experienced an uneventful operative and postoperative course and remains asymptomatic, now more than two years after operation. PMID- 8222846 TI - Dynamic left ventricular outflow tract obstruction. Diagnosis by transesophageal echocardiography in a critically ill patient. AB - Dynamic left ventricular outflow tract obstruction (DLVOTO) can be present in critically ill patients with congestive heart failure. Diagnosis by transthoracic two-dimensional echocardiography may be technically difficult in the critically ill patient or patients who are obese. This report describes the diagnosis of DLVOTO by transesophageal echocardiography and subsequent management. PMID- 8222847 TI - Usefulness of transesophageal echocardiography in patients with hemodynamic deterioration late after cardiac surgery. AB - Two patients with large pericardial thrombi following cardiac surgery presented as having right cardiac tamponade. Transesophageal echocardiography (TEE) identified a large pericardial hematoma compressing the right atrium and was well tolerated by these critically ill patients. These case reports demonstrate the diagnostic value of TEE in the identification of this severe complication in the late postoperative period. PMID- 8222848 TI - Massive macroglossia as a presenting feature of hypothyroid-associated pericardial effusion. AB - An 81-year-old woman presented with massive macroglossia and signs of both hypothyroidism and pericardial tamponade. Drainage of the pericardial effusion produced dramatic resolution of the macroglossia. Marked elevation of central venous pressure may result in macroglossia, possibly because of anomalous venous drainage of the tongue. PMID- 8222849 TI - A nearly fatal tracheal obstruction resulting from a transtracheal oxygen catheter. AB - In a patient with a transtracheal oxygen catheter (ITO2C), a nearly fatal complication occurred due to the formation of a mucous plug on the tip, which almost totally obstructed the tracheal lumen. To our knowledge, this complication has not been reported before with the use of this type of transtracheal oxygen catheter. PMID- 8222850 TI - Behcet's disease presenting with a pulmonary mass lesion. AB - We report a case of Behcet's disease with pulmonary and splenic involvement. The radiologic presentation was quite different from the modes defined in the literature. A large, anteriorly located, well circumscribed ovoid mass was the x ray film image of a large area of infarction with hemorrhage. The involved pulmonary artery showed signs of vasculitis with thrombus formation. To our knowledge, this mode of presentation is different from the cases reported in the literature. PMID- 8222851 TI - Who should perform thoracoscopy? The controversy continues. PMID- 8222852 TI - Who should perform thoracoscopy? The controversy continues. PMID- 8222853 TI - Systemic capillary leak syndrome. PMID- 8222854 TI - Nasal ventilation is not the final word. PMID- 8222855 TI - Neuromuscular blockade in the ICU. PMID- 8222856 TI - Never use neuromuscular blocking agents for sedation. PMID- 8222857 TI - Use of heparin in prevention of venous thromboembolism. PMID- 8222858 TI - Kerley B lines as a radiologic manifestation of Pneumocystis carinii pneumonia. PMID- 8222859 TI - Formal thoracotomy with pulmonary resection. PMID- 8222860 TI - Nebulizers should be labeled accurately. PMID- 8222861 TI - Group C streptococcal pneumonia and aneurysm infection. PMID- 8222862 TI - Pharmacokinetic evaluation of imipenem combined with cilastatin in cystic fibrosis. AB - The pharmacokinetics of imipenem (MK-787) and cilastatin (MK-791) were studied in 30 patients with cystic fibrosis (CF) receiving the drug for therapeutic purposes at doses of 11 mg/kg given as a 30-min infusion. The serum concentrations and urine elimination were studied after the first dose and during steady state. The concentrations were assayed by high-pressure liquid chromatography. The total areas under the imipenem serum concentration curves (AUCs) to infinity were 30.4 +/- 6.8 mg.h/l after the first dose compared with 29.1 +/- 7.1 mg.h/l during steady state (NS). The cilastatin AUCs on the 2 days were 40.3 +/- 9.3 and 38.3 +/- 0.4 mg.h/l (NS), respectively. The urinary recovery of imipenem was 47.8 +/- 17.8% after the first dose and 57.8 +/- 24.2% during steady state (NS). The amounts of cilastatin eliminated in the urine during 6 h were 6/7.3 +/- 22.9% after the first dose and 60.5 +/- 17.0% of the dose during steady state (NS). The mean half-life of imipenem in these CF patients was 1.2 +/- 0.4 h on the first day of the examination and within the same range during steady state. The distribution volume (Vd beta) was in the range of 28 liters, the total body clearance was 16.3 liters/h (285 ml/min) on the first day. The t1/2 of cilastatin was 0.59 +/- 0.14 h after the first dose and 0.61 +/- 0.14 h during steady state. Thus patients with CF eliminated cilastatin more quickly than imipenem. PMID- 8222863 TI - Effects of food, ceruletide and ranitidine on the pharmacokinetics of the oral cephalosporin S-1108 in humans. AB - The pharmacokinetics of S-1108, an ester-type oral cephalosporin antibiotic, were studied in healthy male volunteers in a crossover study. This study focused especially on the effects on S-1108 absorption of (1) a meal, (2) concomitant administration of ceruletide diethylamine (a drug which promotes the secretion of bile) in the fasting state, and (3) concomitant administration of ranitidine hydrochloride (a drug which adjusts the acidity of the gastric juice) in the fasting state. The results show that the absorption of S-1108 after oral administration may be affected by factors such as food and ranitidine hydrochloride. PMID- 8222864 TI - Anti-Brucella activity of Ro 23-9424, a dual-action antibacterial. AB - The in vitro activity of dual-acting antibacterial Ro 23-9424 was determined by the agar dilution method against 126 clinical isolates of Brucella melitensis. It was compared with fleroxacin, ciprofloxacin and five conventional drugs. MIC50 and MIC90 for Ro 23-9424 were 1.0 and 4.0 mg/l, respectively, as compared with 0.25 and 0.5 mg/l for fleroxacin and ciprofloxacin. One strain which was resistant to other fluoroquinolones, with an MIC of > 8.0 mg/l, did not show cross-resistance to Ro 23-9424 and had an MIC of 4.0 mg/l. All the strains were susceptible to conventional drugs like gentamicin, streptomycin rifampicin, tetracycline and trimethoprim-sulfamethoxazole, with MICs ranging between 0.12 and 2.0 mg/l. Ro 23-9424 did not exhibit in vitro synergy with any of the conventional drugs. PMID- 8222865 TI - Comparative stability of meropenem against beta-lactamases from Bacteroides spp. AB - The activity of meropenem was similar to that of imipenem against 88 clinical isolates of Bacteroides spp., including 28 non-fragilis strains. Four of the isolates were able to hydrolyse cephamycins or penems. Investigation of beta lactamases from these strains and 10 laboratory stock culture Bacteroides showed that meropenem and imipenem were degraded at a similar rate by cell-free preparations of these enzymes. Some of these strains were able to completely hydrolyse the carbapenems within 2 h. Despite this rapid hydrolysis, MICs of meropenem and imipenem for some of the strains remained within the susceptible range as judged by agar dilution titration. PMID- 8222866 TI - Killing kinetics of four quinolones against gram-positive cocci. AB - Killing kinetics of four fluoroquinolones against Staphylococcus species were determined during the first 5 h of incubation in the presence of drug concentrations ranging from the MIC to 1,024 micrograms/ml. Additionally, the relationship between killing rate and drug concentration was established. Sparfloxacin and temafloxacin were the most active quinolones assayed with greater lethal rates than ciprofloxacin and norfloxacin. The relation was biphasic with a decrease of at least 1 log10 c.f.u./ml at high drug concentrations with respect to that obtained at the optimal bactericidal concentration for all quinolone-microorganism combinations tested. PMID- 8222867 TI - Mupirocin resistance in methicillin-resistant Staphylococcus aureus from a veterans hospital. AB - 679 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) which occurred from 1986 and 1992 were retrospectively tested to determine the frequency of mupirocin resistance. With disk agar diffusion screening, 26 of 679 MRSA had zones of inhibition of < 18 mm using a 5 micrograms mupirocin disk. Minimal inhibitory concentrations (MICs) by agar dilution on the 26 suspect MRSA revealed that 9 were resistant. Of these 9, 1 had a MIC of 6.25, 4 of 12.5, 1 of 25, 1 of 500 and 2 of > 1,000 micrograms/ml. Although the overall incidence of mupirocin resistance was low in our hospital, 5 of the 9 resistant isolates occurred in 1992 and may signal a much more serious threat in the future. PMID- 8222868 TI - Silver sulphadiazine: a comprehensive in vitro reassessment. AB - The antimicrobial activity of silver sulphadiazine has been determined agonist 409 strains from 12 different genera. MICs were usually in the range 16-64 micrograms/ml. Multi-resistant species, such as methicillin-resistant Staphylococcus aureus and Acinetobacter spp., were uniformly sensitive. MICs always exceeded the reported aqueous solubility of silver sulphadiazine. No resistant strains were found, and there was no correlation between MIC of silver sulphadiazine and resistance to sulphonamides. Time-kill experiments showed that sulphonamide-resistant S. aureus (but not Staphylococcus epidermidis) were killed significantly more slowly than were sulphonamide-sensitive strains. PMID- 8222869 TI - Potentiation of doxorubicin cytotoxicity by the calcium channel blocker verapamil in Ehrlich ascites cells. AB - The calcium channel blocker verapamil increased the intracellular level of doxorubicin in Ehrlich ascites cells. The high cellular drug level was directly related to the enhancement of the cytotoxicity of the antitumor agent. Tumor bearing mice pretreated with verapamil showed a 2.3-fold increase in long-term survival effect of doxorubicin together with a pronounced inhibitory effect on tumor DNA, RNA and protein content. This study suggests the possible novel use of verapamil to enhance the antitumor activity of doxorubicin, allowing its dose, and consequently the serious side effects, to be reduced. PMID- 8222870 TI - A new model examining intracellular and extracellular activity of amoxicillin, azithromycin, and clarithromycin in infected cells. AB - An in vitro infection model was created using a suspension of macrophages, polymorphonuclear leukocytes, lymphocytes, fibroblasts, and human serum to which pathogen and antibiotic were added. Separate intracellular and extracellular antibiotic concentrations and activity against Staphylococcus aureus and Legionella pneumophila were assessed for three antimicrobial agents: amoxicillin, azithromycin and clarithromycin. Amoxicillin was found almost exclusively in extracellular fluid, where it was active; intracellularly, it was ineffective. Azithromycin, in contrast, was primarily concentrated and active intracellularly, with little activity in extracellular fluid. Clarithromycin was present in both compartments and possessed significant activity both intracellularly and extracellularly. PMID- 8222871 TI - In vitro investigation of the intraphagocytic bioactivities of ciprofloxacin and the new fluoroquinolone agents, clinafloxacin (CI-960) and PD 131628. AB - In this study the intraphagocytic bioactivities of the new fluoroquinolone antimicrobial agents clinafloxacin (CI-960) and PD 131628 (the active metabolite of CI-990) were investigated in vitro at final concentrations of 0.0005-0.5 microgram/ml using human neutrophils and the combination of a radiometric and a colony-counting method, which enabled us to distinguish between intracellular bacteriostatic and bactericidal mechanisms. Ciprofloxacin was included for comparison. Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) were used as the test intraphagocytic microbial pathogens. Clinafloxacin (> or = 0.05 microgram/ml) displayed potent intraphagocytic bactericidal activity against S. aureus, while PD 131628 was merely bacteriostatic. Ciprofloxacin displayed relatively unimpressive bacteriostatic rather than bactericidal activity for S. aureus. Against E. coli, the intraphagocytic activity of clinafloxacin (0.001 0.005 microgram/ml and above) was superior to that of PD 131628 or ciprofloxacin, which were approximately equipotent. Clinafloxacin is a potent intraphagocytic bactericidal agent for both gram-positive and gram-negative bacteria. PMID- 8222872 TI - Interference by subinhibitory concentrations of azithromycin with the mechanism of bacterial adhesion to human epithelial cells. AB - Azithromycin is the first member of a novel 15-membered-ring 'azalide' group of macrolides that has entered into clinical practice, and its activity is not restricted to gram-positive bacteria, but extends also to gram-negative bacteria. The aim of this study was to investigate the ability of subinhibitory concentrations (sub-MICs) of azithromycin to interfere with the mechanism of bacterial adhesion to human epithelial cells. Azithromycin induced a significant inhibition of adhesion from 1/2 to 1/32 MIC for Staphylococcus aureus and from 1/2 to 1/16 MIC for Escherichia coli. 1/32 of the MIC for S. aureus means 0.048 microgram/ml, while 1/16 of the MIC for E. coli means 0.25 microgram/ml. At these concentrations no morphological changes in E. coli shape were seen, while sometimes S. aureus cells larger than the normal size appeared. Tissue concentrations of azithromycin decline with an estimated half-life of 2.5-3 days. Since sub-MICs of 0.25 and 0.048 microgram/ml are still able to interfere with bacterial physiology, the effective activity of azithromycin, from a pharmacokinetic point of view, could be extended for 3 days beyond the expected period of antimicrobial activity. PMID- 8222873 TI - Fluconazole in children: first experience with prophylaxis in chemotherapy induced neutropenia in pediatric patients with cancer. AB - In an open multicenter, prospective study fluconazole (FLU) was administered to prevent fungal infection in 22 neutropenic, 2- to 14-year-old children. Of 4 failures, one was infected with an Aspergillus spp., one with Candida krusei and one with Candida (Torulopsis) glabrata. One failure was of bacterial origin. Those patients with mycoses who failed on fluconazole were successfully treated with other antifungals. In 4 patients transient elevation of liver enzymes was observed. This is the first reported trial using FLU in prophylaxis against fungal infection in neutropenic pediatric patients, where 18/24 children did not present any evidence of mycotic infection using FLU in daily doses of 1-4 mg/kg. PMID- 8222874 TI - Efficacy and tolerance of erythromycin acistrate in the treatment of acute exacerbations of chronic bronchitis in the elderly. AB - This randomized, investigator-blind, parallel-group trial compared erythromycin acistrate (EA) and erythromycin base (EB) in the treatment of elderly patients with acute exacerbations of chronic bronchitis. In total, 57 hospitalized patients were included, of whom 28 received EA 400 mg three times daily, and 29 EB 500 mg three times daily for 10-21 days. The mean age of the patients was 70 and 68 years in the EA and EB groups, respectively. The patients underwent medical examination before the onset of the study, at the 7th day during the treatment and 3-5 days after termination of the treatment. The efficacy assessment was based on clinical signs and symptoms of infection as well as on bacteriological culture from sputum samples. 55% of the patients in the EA group and 61% in the EB group were totally cured, 23 and 29%, respectively, had only mild symptoms at the end of the therapy and 14 versus 11% of the patients did not respond at all. The predominant pathogens isolated from sputum were Haemophilus influenzae, Branhamella catarrhalis and Streptococcus pneumoniae. 60% of the patients in the EA group and 46% in the EB group from whom a sample was taken had normal flora in the posttreatment culture. In the EA group, 8 patients and in the EB group 7 patients complained of gastrointestinal side effects and 4 and 1, respectively, discontinued the treatment prematurely. Slight, reversible elevations of one or more liver parenchymal enzyme activities during and after treatment were seen at the same frequency in both treatment groups. The results show that EA is as effective and well tolerated as EB in the treatment of exacerbations of chronic bronchitis in elderly patients. PMID- 8222875 TI - Pulmonary hemorrhage as a clinical manifestation of hemolytic-uremic syndrome associated with mitomycin C therapy. AB - Mitomycin C (MMC) is an alkylating agent that has been recently associated with the hemolytic-uremic syndrome (HUS). Pulmonary impairment in the HUS is rarely reported in the literature, and no reports of pulmonary hemorrhage, as a clinical feature of the HUS, have been documented. We describe two women who developed HUS after MMC therapy and presented massive pulmonary bleeding. Pulmonary hemorrhage is an uncommon feature in the HUS, and seems to appear especially in the HUS associated with MMC therapy. It is extremely resistant to standard treatment, and confers a poor prognosis for this disease. PMID- 8222876 TI - Biological and cultural diversity: the legacy of Darwin for development. AB - To explain becoming human, becoming a member of a culture and society, and becoming a unique human being calls upon diverse theoretical resources in the biological and social sciences. To integrate such diverse concepts requires the umbrella of evolutionary theory, which alone can encompass so many levels of analysis. I present an elaboration of the theory sketched in my presidential address to SRCD in the hope that a more thorough understanding of the theoretical framework and the implications of cultural and biological diversity will enrich and enliven developmental research. Debates with colleagues of different persuasions are always welcome and often fruitful. PMID- 8222877 TI - Genetic change and continuity from fourteen to twenty months: the MacArthur Longitudinal Twin Study. AB - Genetic change as well as continuity was investigated within the domains of temperament, emotion, and cognition/language for 200 pairs of twins assessed at 14 and 20 months of age in the laboratory and home. The second year of life is marked by change rather than continuity: correlations from 14 to 20 months averaged about .30 for observational measures of temperament and emotion, about .40 for language measures, and about .50 for mental development. 2 types of genetic change were examined: changes in the magnitude of genetic influence (heritability) and genetic contributions to change from 14 to 20 months. In general, heritability estimates were similar at 14 and 20 months. Evidence for genetic influence on change from 14 to 20 months emerged for several measures, implying that heritability cannot be equated with stability. Analyses of continuity indicated that genetic factors are largely responsible for continuity from 14 to 20 months. PMID- 8222878 TI - Multiple perspectives on family relationships: a latent variables model. AB - Many scholars are skeptical of family member reports on their interpersonal relationships. Familial reports are assumed to be biased by social desirability as well as other factors. In this study, a latent variables modeling approach was employed to evaluate rater reliability and bias in mother, father, and child ratings of parent-child negativity. Results based on 78 clinical families demonstrate that family member ratings contain a significant "true score" component that correlates with observer ratings of parental behavior. The presence of systematic rater effects is also demonstrated. The latent variables approach, which provides statistical control for rater effects, is recommended for the analysis of this type of data. PMID- 8222879 TI - The interparental relationship and family problem solving with preadolescent males. AB - This study examines associations between the quality of the interparental relationship and how well 68 family triads (mother, father, preadolescent son) solved salient problems which arose at home. Four aspects of the interparental relationship (marital satisfaction, parental agreement, conflict during family problem solving, and parental coalitions) were included in a regression analysis which controlled for family structure and child externalizing. A longitudinal design assessed families when mean child age was 9.7 years and 2 years later. Parental agreement consistently facilitated family problem solving. However, strong parental coalitions inhibited family problem solving, which may be attributed to frustrated autonomy needs of preadolescent males in response to the parental coalition. Stepfamilies had less effective problem solving at Time 1. The results confirm the benefits of parental agreement to child outcomes via enhanced family problem solving but show a reverse effect when agreement occurs in the context of coalitions against a preadolescent son. PMID- 8222880 TI - Maternal support networks, maternal cognitions, and young children's social and cognitive development. AB - The relations among maternal support networks, maternal perceptions of parenting, maternal attributions for parenting situations, and children's social development, as indexed by peer acceptance, and cognitive performance, as indexed by the PPVT-R and PSI, were examined in a sample of 69 mothers and their preschool-age children. Network characteristics directly predicted cognitive performance and indirectly predicted peer acceptance through effects on maternal perceptions and attributions. Parent cognitions as mediators of network effects on children's development are discussed, addressing variation due to network dimensions, types of parent cognitions, and domain of children's development. PMID- 8222881 TI - The relations of emotionality and regulation to preschoolers' social skills and sociometric status. AB - The purpose of this study was to examine the relations of emotionality (intensity and negative emotion) and regulation (coping and attentional regulation) to preschoolers' social skills (as rated by adults) and sociometric status. Teachers' ratings of children's constructive coping and attentional control were positively related to boys' social skills and peer status, whereas negative affect was negatively related. Acting out (vs. avoidant) coping and emotional intensity were negatively related to girls' and boys' social skills and boys' peer status. In addition, mothers' reports of boys' coping by seeking social support and low emotional intensity were associated with boys' positive social functioning, whereas avoidant coping was positively related to girls' rated social skills. The results are discussed in relation to research on emotion regulation and coping with emotion in interpersonal contexts. PMID- 8222882 TI - Biobehavioral organization in securely and insecurely attached infants. AB - Attachment research has shown the emergence of individual differences in the security of infant-mother attachment during the first year of life as well as their importance for later social-emotional development. A biobehavioral perspective may help settle disagreements about the validity and interpretation of 12-month-old infants' different behavioral patterns of attachment assessed by Ainsworth's Strange Situation. It was shown that, despite less overt distress in insecure-avoidant infants after short separations from the mother, overall cardiac measures indicate arousal patterns similar to the secure infants during separation. However, differences in cardiac response emerged with regard to object versus person orientation during reunion. Additionally, findings of increased cortisol in both insecure-avoidant and disorganized infants support the theoretical interpretation that these infants, in contrast to secure infants, lack an appropriate coping strategy. PMID- 8222883 TI - Linking employment to attachment: the mediating effects of maternal separation anxiety and interactive behavior. AB - This study examined the effects of maternal employment and separation anxiety on maternal interactive behavior and infant attachment. 73 mother-infant pairs participated in a laboratory free-play session when infants were 5 and 10 months of age and in the Strange Situation when the infants were 18 months of age. Maternal feelings about being separated from her infant were assessed by questionnaire at 5 months. Employed mothers returned to work before the infants' fifth month, and nonemployed mothers did not work outside the home through their infants' tenth month. Employed mothers who reported high levels of separation anxiety were more likely to exhibit intrusive behaviors at 10 months. While employment was not directly related to attachment, we found infants of high anxiety employed mothers to develop anxious-avoidant attachments. The results suggest that maternal separation anxiety and interactive style may be important mediators between employment and later infant outcome. PMID- 8222884 TI - Family factors related to children's intrinsic/extrinsic motivational orientation and academic performance. AB - This study examined 3 familial factors--parental surveillance of homework, parental reactions to grades, and general family style--in relation to children's motivational orientation and academic performance. Family, parent, and child measures were obtained in the home from 93 fifth graders and their parents. Teachers provided a measure of classroom motivational orientation, and grades and achievement scores were obtained from school records. Higher parental surveillance of homework, parental reactions to grades that included negative control, uninvolvement, or extrinsic reward, and over- and undercontrolling family styles were found to be related to an extrinsic motivational orientation and to lower academic performance. On the other hand, parental encouragement in response to grades children received was associated with an intrinsic motivational orientation, and autonomy-supporting family styles were associated with intrinsic motivation and higher academic performance. In addition, socioeconomic level was a significant predictor of motivational orientation and academic performance. PMID- 8222885 TI - Impression formation in children: influence of gender and expectancy. AB - The effect of expectancy-congruent and -incongruent information on subsequent impression formation and recall of a target peer was examined. 64 boys and 50 girls (ages 8 to 12) were given an expectancy that a stimulus child was either shy or outgoing. Subjects rated the target on several dimensions (e.g., friendly, shy) and then watched a videotape of a boy or girl confederate acting out a script containing both expectancy-congruent and -incongruent information. Subjects then rated the target child again and freely recalled as much of what the target said as they could remember. Results revealed that when children are presented with both expectancy-congruent and -incongruent information, impression formation appears largely attribute based, and the influence of the initial expectancy appears mitigated. However, ratings of liking appeared to be more influenced by the initial expectancy than by subsequent behavioral information. In addition, the results indicated that many of Maccoby's conclusions regarding the effects of gender on social interactions also apply to children's social information processing, indicating a strong bias toward same-sex peers. PMID- 8222886 TI - Young children's long-term retention of a pediatric examination. AB - Children at ages 3, 5 and 7 (N = 187) provided reports of their physical examinations immediately following the checkup and after a delay of either 1, 3, or 6 weeks, or only after 3 weeks. The protocol used for all interviews consisted of initial general prompts, followed by increasingly specific questions as needed. Recall of standard features of the examinations was scored, along with responses to questions concerning acts that did not occur. Retention of the event was extensive and accurate. Although clear age effects were observed in recall, the delay interval did not strongly affect performance over the 6-week period. The amount of forgetting was significant at ages 3 and 5, but not at age 7. With increasing age, a greater percentage of the features was spontaneously recalled. When presented with incongruous questions, all children evidenced rates of correct denials that exceeded chance and few errors of commission. The initial interview did not affect delayed recall. Implications for children's testimony are discussed. PMID- 8222887 TI - Young children's understanding of the mind-body distinction. AB - A experiments investigated children's understanding of the mind-body distinction. Children of ages 4 and 5 recognized not only the differential modifiability of changeable versus unchangeable human properties and bodily versus mental properties, but also the independence of activities of bodily organs from a person's intention (Experiment 1). When presented 3 types of causal explanations (i.e., intentional, vitalistic, mechanical), 6-year-olds chose most often as most plausible for bodily functions vitalistic explanations (i.e., those ascribing the phenomena to a relevant bodily organ's initiative and effortful engagement in activity); 8-year-olds chose the vitalistic explanations second most often, following mechanical ones (Experiment 2). However, 6-year-olds, as well as 8-year olds and adults, did not always choose vitalistic explanations over intentional explanations (Experiment 3); whereas they tended to prefer vitalistic explanations for biological phenomena, they predominantly accepted intentional ones for psychological phenomena (Experiment 3A). These results suggest that children as young as 6 years of age have acquired a form of biology as an autonomous domain which is separate from that of psychology. PMID- 8222888 TI - Clinical features and investigation of traditional Chinese medicine. PMID- 8222889 TI - Exercise therapy of non-insulin dependent diabetes mellitus. Effects of acute exercise loading. AB - The dynamic changes of 15 parameters (divided into 6 sections in this study) relating to metabolism, platelet function, blood coagulation and hemorrheologic situation under acute exercise loading with the intensity of VO2 max 60% were observed in noninsulin dependent diabetes mellitus (NIDDM) patients. The advantage, safety and feasibility of exercise therapy were discussed. PMID- 8222890 TI - Serum isoforms of creatine kinase MM isoenzyme in acute myocardial infarction. AB - Using the electrophoretic method, the changes in the catalytic activities of three CK-MM isoforms in multiple sequential serum samples of 12 patients with acute myocardial infarction (AMI) were monitored for 3 days after the onset of chest pain. In post-AMI period, MM3 reached a peak (518.6 U/L) first, on the average 14.0 hours after AMI, followed by MM2 (19.8 h, 640.5 U/L) and MM1 (28.7 h, 641.3 U/L). According to their faster decay from circulation, MM3 had higher fractional disappearance and appearance rates, followed by MM2 and MM1. The MM3/MM1 activity ratio rose beyond the upper limit, found in health subjects, about 3.5 hours after onset of symptom and peaked on an average 10.6 hours after AMI, even earlier than the peaks of all isoforms, CK and CK-MB. On the other hand, our findings indicated these changes in isoform composition can be of value in estimating the time elapsed since the onset of tissue damage; MM3 is the predominant isoform when the tissue necrosis is of relatively recent origin (5-15 h); MM2 is the dominant subband between 15 and 24 hours after AMI, whereas a predominant MM1 band would indicate a significantly longer time period since the injury occurred (24 hours or more). Thus, these characteristics make CK-MM isoforms an earlier and more sensitive indicator of acute release from necrotic myocardium and an effective mean of predicting the time of the onset of AMI. PMID- 8222891 TI - Therapeutic effects of an androgenic preparation on myocardial ischemia and cardiac function in 62 elderly male coronary heart disease patients. AB - The elevated estradiol/testosterone (E2/T) ratio had been proved to be a risk factor for coronary heart disease (CHD) in elderly men. We conducted a randomized placebo controlled crossover study on the effects of a new androgenic preparation "Andriol" in 62 elderly men with CHD over a period of 2.5 months. The results showed significant differences between Andriol- and placebo-treated groups at the end of this period: in the former, serum T level was elevated significantly (P < 0.001), E2 level was unchanged (P > 0.05), E2/T ratio was reduced (P < 0.05), angina pectoris (AP) was relieved (total effective rate, 77.4%), and myocardial ischemia in ECG and Holter recordings were improved (total effective rate, respectively 68.8% and 75%). Doppler echocardiography showed that 12 parameters of cardiac function were unchanged in both groups. No obvious side effect was found in those who took Andriol. PMID- 8222892 TI - Aberration antigen expression in adult acute myelocytic leukemias. AB - Marrows from 58 newly diagnosed acute myelocytic leukemia (FAB-AML) patients were immunophenotyped by flow cytometry with 13 kinds of monoclonal antibodies. Marrows of 53.4% of the patients showed pure myeloid antigen expression (Ly-AML) and 34.5% displayed both myeloid and lymphocyte associated antigen expression (Ly+AML). In general, fluorescence intensity of lymphocyte associated antigen in Ly+AML was weaker than that of myeloid antigen. Myeloid and lymphoid marker cells distributed randomly in DNA-aneuploidy. It took more days to reach the first complete remission in Ly+AML. The aberration antigen expression in AML allows a sensitive detection of minimal residual leukemic cells in complete remission bone marrow and treatment stratification. PMID- 8222893 TI - Molecular epidemiologic study of an outbreak of Salmonella typhimurium infection at a newborn nursery. AB - Plasmid analysis, restriction endonuclease analysis, antimicrobial susceptibility testing, biotyping, phage typing and outer membrane protein electrophoresis were used to study an outbreak of Salmonella typhimurium infection at a newborn nursery. Seven out of the 12 neonates had positive blood cultures for S. typhimurium, and 2 of them died of severe sepsis. Thirty epidemic strains of S. typhimurium belonging to phage type 12 had the same plasmid profiles (98.0, 6.7 and 3.8 Kb) and identical restriction digest patterns (23.0, 20.4, 15.0, 9.6, 8.2, 7.4, 5.8, 4.3, 3.8, 2.0 and 1.8 Kb) which were different from those of the 2 non-epidemic strains. Laboratory data suggested that the source of the infection was the index patient's mother who had a slight diarrhea; the mode of transmission was most likely due to the transfer of organisms from infant to infant by the contaminated hands of nurses during milk feeding. PMID- 8222894 TI - Epidemiological study on hypothermia in newborns. AB - A study on the incidence and high risk factors of hypothermia in the newborn was carried out in a period of four months (Nov. 1988-Feb. 1989) in 6 counties of 3 provinces. Totally 14,809 newborns were studied, in whom 100 cases were found to be ill with sclerema, having an incidence of 6.7%. Gestational age, body weight, low room temperature, improper methods of thermopreservation during delivery and asphyxia are determined as the most probable high risk factors, in which gestational age and body weight are negatively correlated with the contraction of the disease. It suggests that the prevention of prematurity and asphyxia, and promotion of thermopreservation during delivery should be helpful in preventing sclerema of newborns. PMID- 8222895 TI - Microsurgical recanalization of fallopian tubes after tubosterilization and its related factors. Report of 278 cases. AB - Microsurgical tubocanalization was done in 412 patients. 278 women had the operation before the end of December 1987 and were followed up. The overall intrauterine pregnancy rate was 81.3% (92.86% for isthmus-isthmus anastomosis and 87.5% for ampulla-ampulla anastomosis). Two patients had ectopic pregnancy which was followed by term pregnancy subsequently. The factors affecting operative results were analyzed. PMID- 8222896 TI - Serum SP1, HPL levels and urinary estrogen/creatinine ratio in the detection of low birth weight. AB - Serum pregnancy specific beta 1 glycoprotein (SP1), human placental lactogen (HPL) and urinary estrogen/creatinine (E/C) ratio levels were determined to study their clinical significance in predicting low birth weight (LBW) in women with normal pregnancy and patients with LBW. The results showed that the sensitivity of urinary E/C ratio was better than that of serum SP1 and HPL, and was increased gradually with the reduction of fetal weight. The specificity in predicting LBW decreased gradually with the increase of serum SP1 and HPL levels and urinary E/C ratio. When serum SP1 level was less than 50 mg/L, HPL less than 2.0 mg/L, and E/C ratio less than 10, their specificity could increase over 90%. It is suggested that multi-indexes measurement would improve the accuracy for predicting LBW. We conclude that SP1, HPL and E/C ratio measurement could be an useful index for predicting LBW. PMID- 8222897 TI - Percutaneous lumbar discectomy using a new nucleotome system. Report of 182 cases. AB - One hundred eight-two patients underwent percutaneous lumbar discectomy using a new nucleotome system. The curative and effective rates were 84.6% and 89.6% respectively, without serious complications. To overcome the difficulty of puncture and cutting the nucleus pulposus at L5-S1 interspace, an efficient approach was undertaken to drill through the iliac wing at L5-S1 using a self designed circular drill so that the nucleotome could safely reach L5-S1 disc, the successful rate was 100%. By varying the distance and angle of puncture, it is possible to treat central type of lumbar disc herniation, thereby expanding the indications for this procedure. PMID- 8222898 TI - Organ-association phenomena during sepsis. TNF and IL-6 in different macrophages. AB - Sepsis was induced in rats by cecal ligation and perforation (CLP). Five and 15 hours post CLP, alveolar macrophage (AM), Kupffer cell (KC), and peritoneal macrophage (PM) were isolated and cultured for 18 hours. Culture supernatant was examined for bioactivity of tumor necrosis factor (TNF) and interleukin 6 (IL-6) and response to lipoplysaccharide (LPS) stimulation in vitro. Results showed that AM produced more TNF during sepsis as compared with KC and PM. Stimulation with LPS in vitro was responded only by AM at 15 hr after CLP. Pattern of IL-6 production was different from TNF while KC produced the highest level of IL-6 after induction of sepsis. PMID- 8222899 TI - Streptozotocin induced cardiomyopathy in diabetic rats. AB - Microscopic quantitative cardiomyopathy was previously reported in spontaneous diabetic biobreeding (BB) rats, but the nature of the lesions remains obscure. To further study the specificity of cardiomyopathy in streptozotocin-diabetic rats and discuss the existence of diabetic heart disease, we compared 40 streptozotocin-diabetic rats (D group) with 40 normal control rats (C group) at various durations in plasma glucose, fructosamine, plasma lipid, left ventricular enzyme contents and alterations of myocardial cells and coronary artery tree under electron and light microscope respectively. In 4 weeks, D rats showed myocardial mitochondrial swelling and degeneration, whereas at 8 weeks, myocardial enzyme contents markedly decreased, and myocardial lesions were more conspicuous with disruption of myocardial cells, formation of myocardial contraction bands, dilatation of intercalated discs, deposition of glycogen granules, etc. At 11 weeks, microscopic changes were essentially similar, whereas the decrease of enzyme contents became more conspicuous. During the 11 weeks, persistent and marked hyperglycemia and elevation of fructosamine were observed, however, no abnormalities were found along the coronary artery tree in the D rats. The results indicated that cardiomyopathy is diabetes specific, and exists independently as one of the three important components of diabetic heart disease. PMID- 8222900 TI - Preliminary experience in HDR brachytherapy for 72 nasopharyngeal carcinoma patients. AB - From April 1989 to July 1991, 72 patients with residual or recurrent nasopharyngeal carcinoma (NPC) were treated by MSH Ir-192 brachytherapy or combined external radiation therapy. All were proven histologically. 39 patients with residual disease and 6 with recurrent lesion at the primary site were identified after one or more courses of radical external radiation. Eleven patients had received brachytherapy for boost therapy after one or more courses of external radiation. Sixteen were treated with a planned protocol of external irradiation combined with intracavitary brachytherapy. The overall local control rate was 93.3%; the local control rate for residual and recurrent disease was 100% and 50%, respectively. Soft palate perforation was observed in 3 patients. Brachytherapy is believed to be a useful treatment for residual or recurrent NPC following radical external radiation. PMID- 8222901 TI - Wild-type p53 gene expression in cultured smooth muscle cells from SHR and WKY rats. PMID- 8222902 TI - Partial azurophilic granule deficiency. Report of 2 cases with ultrastructural and light-and-electron-microscopic cytochemical observation. AB - The neutrophils in 2 patients with atopic disease were studied. Ultrastructurally, a number of abnormal azurophilic granules (AG) with low electron-density (Case 1) and secondary lysosomes with "myelinoid membranes" figures (Case 2) were found. Myeloperoxidase (MPO) and acid phosphatase activity were detected by light cytochemical techniques, and showed significantly low values. The deficiency of MPO and abnormal distribution of AG were also demonstrated by electron microscopic cytochemical technique. The neutrophils from the parents revealed changes similar to the patients. This study suggests that since genetic partial deficiency of neutrophil AG enzymes existed, the phagocytosed substances were only partially degraded, leading to accumulation of substances with antigenicity, and became trigger event of atopic disease. PMID- 8222903 TI - Epidemiological features and control strategies of schistosomiasis japonica in China. AB - Schistosomiasis japonica in China has a long history and wide distribution. In the last three decades, great achievements have been obtained in schistosomiasis control. Among 12 formerly endemic provinces, municipalities, and autonomous regions, transmission has been successfully controlled in 2 provinces, 1 municipality and 1 autonomous region. Among 380 formerly endemic counties, transmission has been interrupted in 158 and is under control in 101. Most remaining endemic areas are located in marshlands, lake and mountainous regions. By the ecological and epidemiological features, the endemic areas in lake and mountainous regions can be classified into different categories. Infection is related to exposure to infected lake and ditch water. The inhabitant activities and level of education have much to do with the frequency of water contact. The density of the infected snail is the main index of determining the high-risk place in the mountain and lake regions, and livestock such as buffalo and pigs, the principal reservoir in the endemic areas. Properly arranged annual chemotherapy with praziquantel can reduce the prevalence rate of inhabitants and livestock. However, the prevalence rate can climb again rapidly. Therefore, a maintenance phase is urgently needed. Modification of the snail-ridden environment should be linked with agricultural development and construction of water conservancy. Surveillance should be strengthened in the area where schistosomiasis has been controlled. PMID- 8222904 TI - A brief introduction to a nation-wide sampling survey on schistosomiasis. AB - A nation-wide sampling survey using a stratified group randomized sampling technique was conducted in 1989 in 8 provinces where schistosomiasis was still endemic. Among 343 sampling spots, 197 were in controlled areas and 146 in endemic areas. 16,953 (10.2%) of 165,834 persons were found infected according to stool examination in the endemic areas with a population of 1.03% of the total in China. The positive rates of stool examination in Hunan, Hubei, Jiangxi, Anhui, Jiangsu, Zhejiang, Yunnan and Sichuan provinces were 14.4%, 13.5%, 5.5%, 8.6%, 0%, 0%, 4.4%, and 16.8%, respectively. Thirty-two (0.1%) out of 34,556 children aged 10-14 years were found infected in the areas where schistosomiasis was under control. Also included in this survey were the relationship between schistosome infection and age, sex, and occupation of the population in the endemic areas, the distribution of snail habitats, the roles of domestic animals in the transmission and epidemiological characteristics of different endemic areas. PMID- 8222905 TI - Biology of Schistosoma japonicum. From cercaria penetrating into host skin to producing egg. AB - The biology of the Chinese mainland strain of Schistosoma japonicum in mammalian phase of the life cycle has been studied in the past three decades. The mode of cercarial penetration is found to involve enzyme secretion and movement. The entry of schistosomulum into the blood vessel in the hypodermal connective tissue is evidenced after skin penetration. The growth and development of this fluke in mice and rabbits are investigated by physiological, histochemical and ultrastructural methods. The development of the fluke may be divided into eight stages. In the definitive host-range, at least 45 species and subspecies of 34 genera in 18 families of 7 mammalian orders have been demonstrated, and the susceptibility to infection in domestic animals is compared. The process of egg formation and embryonic development of this fluke are studied histologically and histochemically. It is suggested that the chemical nature of the egg shell should be a quinontanned protein system. The characteristics and dynamic changes of cytochemical substances in embryonation of eggs are described. PMID- 8222906 TI - Diagnostic evaluation of dot-binding assays for circulating cathodic antigen (CCA) and anti-CCA determinations in schistosomiasis japonica using defined biotinylated conjugates. AB - Using an affinity purified circulating cathodic antigen (CCA) preparation and an anti-CCA monoclonal IgM antibody, we modified dot-binding assays for anti-CCA and CCA detections with biotinylated conjugates. A study on 3 groups of 138 schistosomiasis japonica patients and 105 healthy individuals demonstrated a high predictive rate of over 95% in both assays, highly comparable to those of circumoral precipitin test (COPT). A blind test in 342 samples of various groups revealed higher detection rates in Ab-binding assay with both acute and chronic case groups (over 90%), but comparatively lower rates in Ag-binding assay with chronic groups (50%-76%). Distinct reductions of either GMRT and dot-indexes were found in 48 praziquantel treated patients whose sera were collected 9 months after chemotherapy. The major target molecules detected by the two binding assays were proved to be the protein incorporated moieties readily precipitated by trichloroacetic acid (TCA), ammonium sulphate and higher concentration of polyethylene glycol (PEG) suggestive of pathological, specific immunoglobulins in the free or complex forms. Non-specific dot reactions were found in some acute and chronic patients with non-relevant bovine serum albumin (BSA) conjugated peroxidase and streptavidin-PO controls, and also in normal sera with the McAb peroxidase conjugate or when working concentration of the biotinylated McAb was not properly titrated. The reliability and proper management of the dot binding assay as an immunodiagnostic tool were discussed. PMID- 8222907 TI - Levo-praziquantel versus praziquantel in experimental and clinical treatment of schistosomiasis japonica. AB - Levo-praziquantel is the left isomer of racemic praziquantel. Animal experiments showed that it is an active component of schistosomicidal activity, while dextro praziquantel is almost ineffective. Clinical trials in three endemic areas of Schistosomiasis japonica indicated that the therapeutic efficacy of levo praziquantel is superior to racemic praziquantel. Pharmacokinetic and pharmacodynamic activities of the stereoselectivity of praziquantel isomers (enantiomers) are discussed. PMID- 8222908 TI - Increased lipoprotein (a) as an independent risk factor for cardiovascular and cerebrovascular diseases. AB - We quantified serum lipoprotein(a) [Lp(a)] in 1266 healthy persons and established the reference value of Lp(a) in groups of age and sex. The Lp(a) in myocardial infarction (MI) survivors, stroke and primary hypertension patients were determined in comparison with that of age-matched normal controls. Moreover, cholesterol, triglyceride, HDL-cholesterol, apo Al and apo B were measured. The Lp(a) concentrations were not correlated with other lipids in normal controls and patients. Our results suggest that increased Lp(a) is an independent risk factor for MI and stroke patients. The total detective rate of abnormal lipids in MI and stroke patients was as high as 50% and 53.8% (Lp(a) excluded) as well as 65.7% and 64.9% (Lp(a) included) respectively. PMID- 8222909 TI - The relationship between cytosolic free [Ca2+]i concentration and intracellular cAMP of platelet at various stages of experimental atherosclerosis in rabbits. AB - The relationship between cytosolic free [Ca2+]i and intracellular cAMP of platelet at various stages of experimental atherosclerosis in rabbits was studied. The results showed that the increase of [Ca2+]i was negatively correlated with the level of cAMP in cholesterol-fed rabbits (r = -0.667, P < 0.001) for 2, 4, 8 and 12 weeks. The viscosity of the whole blood at shear rates of 230.4/s and 5.76/s increased only in rabbits fed with cholesterol for 2 weeks, but the plasma viscosity increased in those fed for 2 and 4 weeks. PMID- 8222910 TI - Safety and immunogenicity of live attenuated hepatitis A virus vaccine (H2 strain) in humans. AB - A study was made in Neijiang City, Sichuan Province to observe the safety and immunogenicity of a live attenuated hepatitis A virus (HAV) vaccine (H2 strain). 3,031 children, aged 6 to 9 years, were injected subcutaneously with 10(6.0) TCID50, 10(5.5) TCID50, and 10(5.0) TCID50 of HAV vaccine; the seroconversion rate and antibody GMT were 97.52%-90.68%, GMT = 1:5.19 +/- 1.90-1:3.36 +/- 1.94; 84.75%-76.27%, GMT = 1:4.29 +/- 1.96-1:2.0 +/- 2.67; 53.23%-51.61%, GMT = 1:2.57 +/- 1.89-1:1.68 +/- 1.41 respectively after 4 weeks to 12 months of inoculation. Forty-eight children in the control group were anti-HAV negative at 4, 8, and 12 weeks; 21 children in the oral group were not antibody positive 4 and 8 weeks after oral administration of the vaccine. The stool specimens of 3 of the 14 vaccinated children were HAA positive. The results suggest that the live attenuated HAV vaccine (H2 strain) is of good immunogenicity and safety. PMID- 8222911 TI - DNA flow cytometry from paraffin-embedded tissue in diagnosis of cutaneous malignant lymphomas and pseudolymphomas. AB - DNA content and cell cycle distributions in paraffin-embedded blocks of 111 skin biopsy specimens of 70 patients with cutaneous malignant lymphomas (CML) and 41 patients with cutaneous pseudolymphomas (CPL) including chronic actinic dermatitis (CAD) were estimated by DNA flow cytometry. A statistical significant difference between DNA indices (DIs) or proliferative indices (PIs) for CML including mycosis fungoides (MF) II, MF III, Sezary's syndrome (SS), cutaneous peripheral T-cell lymphomas other than MF and SS, cutaneous germinal center cell derived lymphomas and cutaneous genuine histiocytic lymphoma from CPL. DIs were also helpful in differentiating MF I from CPL. There was a linear relationship between DIs and PIs, both of which had a parallel relationship to the degrees of malignancy and mortality of varieties of CML. The finding of aneuploidy is likely to be useful in differentiating CML from CPL. It is worth noting that DIs, PIs and proportions of aneuploidy in CAD were all higher than those of higher malignancy of CML. These data could not be considered as markers of malignancy. PMID- 8222912 TI - The effect of reconstruction after subtotal gastrectomy on release of vasoactive intestinal peptide. AB - To avoid dumping after Billroth gastrectomy, we designed pylorus and antroseromuscular-flap preserving subtotal gastrectomy (PAFPG). The mean maximal plasma level of VIP (vasoactive intestinal peptide) in PAFPG after oral hypertonic glucose was close to that in the control. Compared with PAFPG group, the plasma VIP concentrations after hypertonic glucose ingestion in BI and BII group were significantly higher, and the concentration of plasma VIP in dumpers was significantly higher than that in non-dumpers after BI or BII gastrectomy. The results suggest that the amount of VIP release is related to the type of reconstruction after gastrectomy. PMID- 8222913 TI - Immunodiagnosis of Schistosoma japonicum infection in China. AB - Diagnosis is an essential prerequisite for the control programs of schistosomiasis. Chinese scientists have been making efforts for decades towards the development of immunological approach in an attempt to offer alternatives of or supplements to parasitological diagnosis. The authors outline briefly the main advances in the research of immunodiagnosis for schistosomiasis japonica in China in two catalogues: antibody detection and antigen detection. A number of immunodiagnostic assays based on antibody detection, which have been used widely in the control programs in China, are appraised in terms of sensitivity and specificity, and the achievements in the development of immunodiagnostic assays based on antigen detection are introduced. Readers will find some information concerning the efforts so far made by Chinese scientists in the improvement of immunodiagnosis by use of molecular approach, including search for biochemically and/or immunochemically isolated and/or purified antigens, recombinant antigens and anti-idiotypic reagents of diagnostic interest to construct defined antigen antibody systems for diagnosis, and development of monoclonal antibody-based assays for antibody or antigen detections. PMID- 8222914 TI - [The existence of the private established surgeon is threatened. Catastrophic development of economic principles of ambulatory surgery--fees for ambulatory surgery]. PMID- 8222916 TI - [The legal pension status of surgeons of former East Germany--a written exchange with the federal employment administrator]. PMID- 8222915 TI - [Surgery and the surgeon in self reflection]. PMID- 8222917 TI - [Parks ambulatory hemorrhoidectomy]. PMID- 8222918 TI - [Ambulatory surgery]. PMID- 8222919 TI - [Ambulatory surgery in practice]. PMID- 8222920 TI - [Is laparoscopic cholecystectomy justifiable as an ambulatory intervention?]. PMID- 8222921 TI - [Comment on the article "Pilonidal sinus" by G. Mehlke]. PMID- 8222922 TI - [Services of surgeons to other clinics]. PMID- 8222923 TI - [Molecular principles of carcinogenesis. Significance for prevention and early detection of solid malignant tumors]. PMID- 8222924 TI - [Gene therapy of malignant tumors]. PMID- 8222925 TI - [Anastomosis techniques for the upper intestinal tract]. PMID- 8222926 TI - [Anastomosis techniques for the lower intestinal tract]. PMID- 8222927 TI - [Results of preoperative radio-chemotherapy in locally advanced squamous epithelial cancer of the esophagus]. AB - In a 4-year period (1988-1991) 122 patients with a squamous cell carcinoma of the esophagus were studied prospectively and analysed. 64 patients of them could be primary resected (primary resectability rate 52%), 36 patients were in general inoperable and 22 patients had an advanced stage of cancer with local inoperability. Due to a preoperative combined radiotherapy and chemotherapy 16 of the 22 patients with local inoperability had a clinical remission of the tumor (73%). 11 patients (50%) showed a histological verified down staging and 3 cases of them a complete remission (no primary tumor was found, no infiltration of the regional lymphnodes and no metastatic disease). Curative resection was possible in 14 of 16 patients with clinical remission (2 patients refused surgical treatment). So the resectability rate now increases from 52 to 63%. We conclude that there was no increased rate of postoperative complications or mortality in the combined radio-/chemotherapy group compared with the primary resected patients. PMID- 8222928 TI - [Long-term follow-up with various Dacron bifurcation prostheses. 5-years results of a prospective randomized study]. AB - In the frame of a prospective randomised study various Dacron bifurcation prostheses with or without velours or with or without gelatin impregnation were implanted in one hundred patients. Fifty-four patients could be reexamined clinically and sonographically after an average of 66 months. A total of 23 patients had died during the period of observation. A comparison of the Fontaine stages during reexamination indicated in 72.3% of cases an operation-related improvement as compared to the condition in the preoperative stage. Sonographically, an average dilatation of 17.3% could be proved. A significant difference due to material or impregnation was not observed. After 72 months the limb salvage amounted to 83.0%. A comparison of the various types of prostheses regarding their limb salvage could indicate no significant difference in the course of 72 months. Based on the only slightly differing results within the period of observation, only the more comfortable intraoperative handling leads to giving preference to the use of double velours prostheses in comparison to prostheses without velours. A positive influence of impregnation with gelatin could not be established. PMID- 8222929 TI - [Heparin-associated thrombocytopenia as a problem in heparin therapy]. AB - The clinical picture of HAT (heparin-associated thrombocytopenia) is being described in reference to a typical case report. If not recognized in time, HAT may lead to serious thrombo-embolic complications, which may eventually lead to death. Considering the wide applications of routine heparin in therapy and prophylaxis, HAT deservers more clinical attention. Routine monitoring of platelet counts during heparin treatment is crucial and essential. PMID- 8222930 TI - [Diagnosis of anorectal fistula using magnetic resonance tomography]. AB - In 17 patients with anorectal fistulae the pelvis was examined by magnetic resonance imaging (MRI). The extent and the course of the fistulae and abscesses could be visualized by proton-density- and T2-weighted images. In 6 of 17 patients fistulae were seen that could not be detected by clinical investigation. In 2 cases additional pelvirectal abscesses were found. Most of the fistulae (5 of 6) had a complicated high suprasphincteric or rectovaginal course. It is concluded that with MRI anorectal fistulae and abscesses can be visualized and that this method is a worthful additional diagnostic tool in patients with complicated fistula-in-ano. PMID- 8222931 TI - [Surgical technique in interventions on the thyroid gland]. AB - The thin fascial layer covering the vessels of the throat and the recurrent laryngeal nerve represents an essential structure in thyroid surgery. For protecting the parathyroid artery and the recurrent laryngeal nerve this fascia must be dissected away from the thyroid capsula propria. Incomplete dorsal dissection may leave dorsal nodes leading to recurrent goiter or recurrent autonomy, whereas penetrating the fascia may injure the recurrent laryngeal nerve and the parathyroid glands. This various handling of the covering fascia can be traced from Billroth and Kocher until today. It determines also whether the recurrent laryngeal nerve is dissected free early and completely during thyroidectomy or whether it is only located behind the fascia or through a small window. PMID- 8222932 TI - [Interdisciplinary management in extremity saving resections of lower extremity tumors]. AB - Eighteen patients with tumors of the leg underwent limb saving operations. The large tissue defects were covered by flaps. The use and advantages of these flaps are discussed. PMID- 8222933 TI - [Successful embolectomy of the main branches of the celiac trunk and the superior mesenteric artery]. AB - This is a case report of a female patient, 81 years old, with embolic occlusions of the celiac trunk and superior mesenteric artery. The surgical therapy was embolectomy of both vessels through a transversal arteriotomy. The peripheral mesenteric vascular bed was flushed with a special conservation solution in continuity. It is given a plentiful discussion about the details of this successful treated course of disease. PMID- 8222934 TI - [Esophageal duplication in a former bronchogenic cyst. Case report and review of the literature]. AB - We report a case of a cystic esophageal duplication with primary cardiac symptoms and bronchogenic cyst in an adult. Duplications of the esophagus are often operated on the diagnosis of mediastinal tumor. With new imaging techniques the diagnosis can be guessed before operation. Nuclear magnetic resonance and endoluminal ultrasound are two newer techniques which are valuable in assessing "mediastinal" tumors. PMID- 8222935 TI - [Para-esophageal omental incarceration--a rare complication of para-esophageal hernia]. AB - Herniation of large omentum is a rare complication of parahiatal hernias. This paper presents by means of a case report clinical symptoms, radiologic procedure and operative management of parahiatal greater omentum incarceration. PMID- 8222936 TI - [Ectopic thyroid gland tissue in the liver]. AB - Heterotopia of thyroid gland can be seen very seldom in humans. Most frequently ectopic thyroid tissue is found in the middle line of neck and chest. Ectopia of thyroid tissue caudal of the diaphragm is an exceptionally rare finding. An explanation results out of the embryology of the thyroid gland development. Reviewing the literature we could find only one case of accessory thyroid tissue in the region of the hepatic porta, it was found at autopsy. In the present case report we describe a 58-year-old woman who had an indistinct tumor in the hepatic porta which was successfully removed. Histologic findings was that of aberrant normal thyroid tissue. PMID- 8222937 TI - [Evaluation of the panoramic sialography in the diagnosis of salivary gland diseases]. AB - Pantomographic sialography (PS) and traditional sialography (TS) was performed and compared on 106 glands out of 94 patients. The main advantages of pantomographic sialography are: (1) sialectasic lesions of the salivary glands are shown well; (2) small lesions of the main salivary glands are shown well: (3) lesions of the main salivary ducts are shown well; (4) the imaging distortions are minor and the actual situation can be demonstrated well; (5) superimposition of surrounding tissues is avoided, the imagings are more clear; (6) more than one glands can be examined at the same time; and (7) patients are positioned easily and feel more comfortable. PMID- 8222938 TI - [Morphological changes in tongue cancer after cryosurgery]. AB - Tca 8113 (human tongue cancer cell line) cell transplanted tumors in nude mice were treated with cryosurgery for three freeze-thaw cycles. Tumor samples were obtained by biopsies pre- and post-cryosurgery for morphological study. The results showed intercellular adhesion damage, nuclear pyknosis, cell death, etc. One week after, the deep parts of the frozen samples were similar to that of the untreated ones. Our study indicates the change of biomembrance may be also important as of nuclei in cell death and may play an important role in the treatment of cancer by cryochemistry. PMID- 8222939 TI - [Application of porcelain veneers]. AB - This paper introduces the technique of porcelain veneer restoration. The retention, cosmetic effect, gingival response, type of tooth preparation and stain of porcelain veneers were discussed. 12-20 months following clinical observation of 92 cases has been undertaken. It is concluded that porcelain veneers offer the advantages of aesthetics, stain resistance, wear resistance, colour stability and soft-tissue compatibility. PMID- 8222940 TI - [Clinical evaluation of the ultrasonic root canal instrumentation]. AB - Five hundred and fifty-nine teeth from 432 patient' were divided randomly into the ultrasonic or the hand root canal instrumentation group. The results are shown as follows: Less instrumentation time was needed significantly in the ultrasonic group (anterior tooth: 2.37 +/- 1.37 minutes, posterior tooth: 5.33 +/ 2.83 minutes) than in the hand group (anterior tooth: 3.15 +/- 2.43 minutes, posterior tooth: 8.42 +/- 4.64 minutes); The rate of instrument fracture in the ultrasonic group (5.28%) was significantly lower than in the hand group (10.97%); No significant difference both in the rate of postoperative pain within 1 week after root canal instrumentation and the 1 year treatment effective rate were found between two groups. PMID- 8222941 TI - [The functional occlusal contact area and masticatory efficiency in complete denture]. AB - The authors developed a new method to measure the occlusal contact area in complete denture by using the IBAS 2000 (Interactive Image Analysis System). The masticatory efficiency and functional occlusal contact area were measured and analysed for 16 cases of complete dentures. The results showed that the relation between occlusal contact area and masticatory efficiency was positive significantly and different region of occlusal contact area had its corresponding masticatory efficiency. It is indicated that a given occlusal contact area is necessary to satisfy the masticatory efficiency required. If the masticatory load is great the width of supporting cusp should be reduced and a definite width of nonsupporting cusp should be preserved for holding the food being masticated. PMID- 8222942 TI - [Oral implantation with HA-Ti-Implant]. AB - The HA-Ti-Implant is a cylindrical device with titanium as kernel and a HA-coated surface. In the clinical use, a two-stage surgical procedure is recommended. In the past 1.5 years, a total of 98 implants inserted in 63 patients were performed, including 64 immediate and 34 delayed implantation. On 67 implants, dental crowns and bridges were connected. Only one implant failed in osteointegration during the healing phase. 34 implants have been followed-up for more than one year. PMID- 8222943 TI - [The maxillofacial injury associated with multiple trauma: a prospective study of diagnostic miss in fractures]. PMID- 8222944 TI - [Sialography with the time-pressure curve monitors]. AB - A new procedure was established for sialography that it is monitored by the time pressure curve during automatically continuous infusion of Urografin 76%. Shape of the curves in 10 normal parotid glands shows firstly the injection pressure rises to the highest peak rapidly, then falls slowly. Eventually the pressure remains constant despite continued infusion. At that time, mean pressure is 7.3kPa (range is 5.3-9.3 kPa). 3 radiographs are taken at 1 minute intervals from the time of the injection pressure beginning to keep constant. The result is that the second sialogram image is the most satisfied. PMID- 8222945 TI - [Clinical observation of treatment of tooth hypersensitivity with silver ammonia fluoride solution]. PMID- 8222946 TI - [Prevalence of dental fluorosis in Fangcun, Guangzhou and Hong Kong fluoridated to 0.7mg/L]. PMID- 8222947 TI - [Cell proliferation in oral candidal leukoplakia]. AB - Epithelial thickness, mitotic activity and DNA synthesis in oral candidal leukoplakia (chronic hyperplastic candidosis) were studied. The results showed that the epithelium was thicker, the number of mitosis and DNA synthesis cells were more in candidal leukoplakia than in non-candidal leukoplakia. These findings indicate an active cell proliferation exists in candidal leukoplakia which may be a precancerous lesion. PMID- 8222948 TI - [Multiple comprehensive analyses of immune regulate factors in patients with oral lichen planus]. PMID- 8222949 TI - [An observation of the effects of rat's condylar cartilage after application of retractive force on the mandible]. AB - The changes of histologic structure and matrix secreting of rat's condylar cartilage after application of upward retractive force to the mandible were investigated by means of histology and histochemistry. After the force application, the condylar cartilage became thinner, the amount of the cells in the germ zone decreased, and that in the transition and transform zone were extremely or even disappeared. Sometimes a calcification zone was discovered in the cartilage and some cells of the mature zone were embed in it. The amount of acid mucopolysaccharide in the matrix also decreased. The result indicated that the retractive force inhibited the proliferation and differentiation of condylar cartilage remarkedly. This was the mainly morphologic bases of the shorted mandible which as a result of the retractive force applied to the mandible. PMID- 8222950 TI - [A study of temporomandibular joint with retruded closed position of edentulous jaws by CT and MRI]. AB - Bilateral TMJ of 7 edentulous patients were studied by CT in two positions. One position was to make the complete dentures in intercuspal position (ICP) while the other one was to make the edentulous jaws in closed position. Two of them MRI examination was made. It is discovered that the condyles move backward and upward in the glenoid fossa. The displacement of the disc in edentulous closed position is less in comparison with ICP. It is proposed that loss of vertical dimension such as abrasion and loss of dentition may press the disc anteriorly as a result of condyle displacement which it is a causal factor of TMJDS. PMID- 8222951 TI - [The relation between the resistance distribution on crown surface and caries]. PMID- 8222952 TI - [An immunobiochemical study of saliva in children with dental caries]. PMID- 8222953 TI - [Stress analysis of dentes incisivi and membrane alveolodental with clinical significance]. PMID- 8222954 TI - [Radiological study on jaw lesions in patients with histiocytosis X]. AB - Thirty two cases of histiocytosis X were collected in this present study. The posterior region of the mandible was the predominant site in histiocytosis X with jaw involvement. Jaw lesions could be divided into two types: 1. Type I (Alveolar Type)--the bone was destroyed along the alveola. This type is seen more frequently in the disseminated cases than in the eosinophilic granuloma of bone (EGB), and the majority of the lesions in this type manifested as ulcerative lesions clinically; 2. Type II (Jaw Body Type)--the mandibular body was destroyed in two ways which manifested as bony radiolucence with or without partitions. Most of the lesions had local swelling or massive manifestations clinically, and almost all lesions occurred in mandible, particularly in the cases of EGB. PMID- 8222955 TI - [Expert system for the design of removable partial denture]. AB - After analyzing the knowledge of the design of R.P.D. and the prosthodontist's general thinking modules, we may divide the knowledge concerning partial denture design into four levels: Domain, Inference, Task, Tactics and use the most mature method of production rules to express various design theories, principles and experiences. By the method of inference, use relevant prosthodontic rules in accordance with the positions and number of lost tooth of the actual cases and getting the relevant result of the design, it is differ from the general date base system of R.P.D. This system is realized on PC-computer and take up about 35 KB, it might accommodate all possible type of deficiency combinations, that is 65,534 in the upper and lower partial edentulate arches and relevant schemes of R.P.D. PMID- 8222956 TI - [Diagnosis of the infratemporal fossa neoplasm by CT]. PMID- 8222957 TI - [Observation of ultrasonic effect on orthodontic tooth movement through transmission electron microscopy]. PMID- 8222958 TI - [Pigmented nevi of the oral mucosa]. PMID- 8222959 TI - [Detection of plasma thromboxane B2 and 6-keto-prostaglandin Fl alpha in patients with recurrent aphthous ulceration]. PMID- 8222960 TI - [Biocompatibility of Cu-Zn, Ni-Cr, Co-Cr alloy]. PMID- 8222961 TI - [The characteristics of temporomandibular joint syndrome in the elderly and a primary observation of the treatment with splint]. PMID- 8222962 TI - [Changes in hemodynamics after ligation and embolization of the external carotid artery]. AB - The authors were the first ones who obtained the established collateral vascular anatomy after external carotid artery embolization and ligation with angiography, vascular transparent specimen filled with ink, microvessel casting and scanning electronic microscope. The results indicated that the rich collateral circulation would be established after external carotid artery ligation. Ligation itself could not cut off blood supply to lesion area. Embolization, in turn, can effectively obstruct blood supply by reducing collateral branches. So we suggest that it is better to use external carotid embolization rather than external carotid ligation for those oral maxillofacial lesions with rich blood supply. PMID- 8222963 TI - [Role of cellular factors in degenerative osteoarthritis]. PMID- 8222964 TI - [Evaluation of the normal adults' condylar position in the fossa]. AB - 44 normal adults were strictly selected from 450 subjects. Fully corrected lateral tomograms and individual corrected transcranial radiograms were taken in the intercuspid position. The area and linear measurements of the joint space were performed with the aid of a computer. 70 TMJs of the fully corrected lateral tomograms were included in the present study. The measurements of joint space area showed that the normal condylar position is in the center of the fossa basically. However, obvious variations of the condylar position could be seen. Therefore, the concentric position was not the only normal physiological position of the condyle. PMID- 8222965 TI - [Effect of interleukin-1 beta on liver/bone/kidney alkaline phosphatase mRNA level of fibroblastic cells derived from human periodontal ligament]. AB - The present study showed the effect of interleukin-1 beta (IL-1 beta) on liver/bone/kidney (L/B/K) alkaline phosphatase (ALP) mRNA level of the fibroblastic cells derived from human periodontal ligament (HPLF). The 5th to 10th passage of cultured HPLF were used. It was found that IL-1 beta significantly inhibited the gene expression of L/B/K ALP mRNA in the cells, and that the effect was dose-and incubation time-dependent. These results suggest that IL-1 beta may play a negative role in reparation of periodontium and periapical diseases. PMID- 8222966 TI - [Evaluation of relation between signs of trauma from occlusion and severity of periodontitis]. AB - The aim of the present study was to evaluate the various occlusal interferences, signs of trauma from occlusion and their relations to severity of periodontitis. 32 cases of moderate to advanced adult periodontitis patients participated in the study with a mean age of 37.6 years. All subjects received a complete periodontal examination and occlusal analysis. The results indicated that teeth with various occlusal interferences did not exhibit any greater severity of periodontitis than those without interferences. However, widened periodontal ligament space (PDLS) was frequently seen in teeth with occlusal interference. Two combined criteria were initially suggested to identify the occlusal trauma in clinic. The first criterion included presence of functional tooth mobility plus radiographically widened PDLS. The second, presence of significant tooth wear plus radiographically thickened lamina dura. The results showed that teeth with the first criteria had deeper probing depth, more clinical attachment loss and less alveolar bone support than teeth without these findings. However, teeth with the second criteria had less attachment loss and more bone support than teeth from control group. It was suggested that both of the combined criteria provide better reliability in determination of established occlusal trauma as well as evaluation of adaptive capacity of periodontal structure to increased biting force. The regression analysis demonstrated that given equal clinical attachment levels and similar inflammation, teeth with evidences of functional mobility and widened PDLS had less osseous support than teeth without these findings (P < 0.01). The difference became greater in line with the more loss of clinical attachment. PMID- 8222967 TI - [Scanning electronic microscope (SCM) examination of the hydroxylapatite (HA) combined with bone morphogenetic protein (BMP) implanted on the bone surface]. AB - HA combined with BMP was implanted on the surface of rabbit's parietal bone and examined with SCM. The results indicate that there are a large amount of new bone formation within the combined material and the amount is increasing as the time lapsed. The new bone is directly connected with the HA particles, forming a bony interface between the host bone and the implants to integrate them. In contrast, there are a lot of fibrous tissues with a little new bone formed on the interface if HA is implanted alone. The result implies that the BMP plays a very important role in new bone formation and the combined material is ideal for alveolar ridge augmentation. PMID- 8222968 TI - [The relation between contour and stress of metal-resin interface]. AB - The interface residual stress is closely related to the interface contour of metal-resin, which affects the service life of metal-resin restoration. The authors researched the relationship between the interface residual stress of Ni Cr alloy-acrylic resin and the different interface contours such as the curvature, convexity, concavity, width and the angle of interface. The result showed that the stress analysis with the Laser-optical microscopic analyzer is an advanced method to study metal-resin interface stress. The interface contour of concave, shallow cavity without sharp margin is desirable for reducing the interface stress. PMID- 8222969 TI - [Electroencephalographic study of bruxers during sleep]. AB - A whole night recording of EEG, EMG and EOG of 7 bruxers during sleep with S-EEG equipment is made. Two of them have been evaluated after splint therapy. After analysing the sleep architecture, sleep progress and sleep index, it is found that: (1) S2 period is shorter, REM period is longer and the REM percentage is higher in comparison with the control group; (2) The total teeth grinding period is 213-691 secs and the frequency is 24-70 times per night, the occurrence of teeth grinding is scattered in different sleep stages; (3) The frequency and period of teeth grinding are decreased significantly after splint therapy; and (4) The occurrence of teeth grinding is a kind of functional disturbance of central nervous system rather than. The aroused response or transfer of sleep stages. PMID- 8222970 TI - [Sagittal fracture of mandibular condyle (SFMC): a clinical and pathological study on the operated patients]. AB - 8 patients with SFMC (9 TMJs) were treated surgically, of which findings on images, during operation and pathological examination demonstrated that SFMC of human being was able to reach bony healing, however, with a larger transverse diameter and a step-like articular surface or a bifid condyle; and that both the fractured and the remained condylar cartilage early show degeneration deteriorating progressively, as a result, all 3 six-month or more SFMC patients had perforation of disc and adhesion in TMJ and complained of TMJDS symptoms. These results support the views that the fate of SFMC is severe secondary osteoarthrosis and that SFMC patients should be treated with positive procedures as early as possible. PMID- 8222971 TI - [Quantitative analysis for mast cells in obstructive sialadenitis]. AB - Quantitative analysis for mast cells in 27 cases of obstructive sialadenitis, 12 cases of approximate normal salivary gland tissues and 5 cases of lymphoepithelial lesion of salivary glands shows that the number of mast cells is slightly increased with the increase of gravity-grade of obstructive sialadenitis and this is closely related to fibrosis of salivary glands and infiltration grade of inflammation cells (dominated by lymphocyte cells), whereas not closely relating to the age change of patients. For the cases of benign lymphoepithelial lesion of salivary glands with malignant changes despite of malignant lymphoma or squamous cell carcinoma the numbers of mast cells are obviously decreased. PMID- 8222972 TI - [Relationship of pathomorphological classification of muco-epidermoid tumor and DNA content]. PMID- 8222974 TI - [The nomenclature of Helicobacter pylori]. PMID- 8222973 TI - [Immunogenicity of deep-frozen allogenic bone transplantation]. PMID- 8222975 TI - [In vitro antibacterial activity and clinical significance of domestic fluoroquinolones combinations with other antimicrobial agents]. AB - The minimal inhibitory concentration of norfloxacin, pefloxacin, ciprofloxacin and other ten antimicrobial agents for 143 strains of Gram-positive cocci and 267 strains of Gram-negative bacilli of recent clinical isolates from June 1990 to March 1991 was analyzed. The results showed that ciprofloxacin and norrancomycin were more active than other antimicrobial agents against most of the Gram positive cocci, while ciprofloxacin and the third generation cephalosporins were more active than other antimicrobial agents against most of the Gram-negative bacilli. The reason for the resistance to antimicrobial agents may be associated with the fact that these bacteria were isolated from inpatients with chronic urinary tract infection or lower respiratory infection of preexisting chronic pulmonary diseases, who had been treated with many antimicrobial agents. Bacteria with resistance to pefloxacin and ciprofloxacin have been found, though these agents have never been used in Tianjin. Resistance of bacteria to ciprofloxacin and pefloxacin may be concerned with cross-resistance that resulted from prolonged and widespread use of norfloxacin in Tianjin. It was also found that norvancomycin resistance (MIC, > 16 mg/L) was encountered in 6 of 34 clinical isolates of oxacillin-resistant staphylococcus aureus. PMID- 8222976 TI - [Identification of latent HBV infection in primary hepatic carcinoma and liver cirrhosis with polymerase chain reaction]. AB - In order to explore latent. HBV infection in primary hepatic carcinoma (PHC) and liver cirrhosis (LC), serum HBV X and S gene were tested with polymerase chain reaction in 40 cases of PHC and LC with both negative HBsAg and HBeAg. The positive rates for X and S gene were much the same. The S gene-positive cases were, however, slightly more than the X gene-positive ones. In the cases of PHC and LC with all the five serum immunologic markers negative HBV DNA was found in 56.3% (9/16) and 45.5% (5/11) of the cases respectively. Of those patients with positive anti-HBs or anti-HBe, HBV DNA was found in 4/8 and 1/4 of the cases respectively. It was shown that in the cases of PHC and LC with all the five markers-negative, about half of them seems to be still related with HBV infection. Some of the PHC and LC cases with positive anti-HBs or anti-HBe may also have low level of viral replication. Detection of HBV DNA with multiple genes is of help to identify more PHC and LC patients with latent HBV infection. PMID- 8222977 TI - [The twenty-year trend in hospital mortality of acute myocardial infarction in Shanghai and the responsible factors. Shanghai Cooperative Group of Myocardial Infarction]. AB - The hospital mortality rates of patients with acute myocardial infarction admitted to 10 main hospitals in Shanghai during the period of 1970-1989 were compared with those of different time periods (year 1970-1974, 1975-1979, 1980 1984 and 1985-1989) and the results disclosed a decreasing trend in mortality in later years. The distribution of major prognostic factors were compared among patients admitted in different time periods and no trend to naturally reduced infarct severity on admission was found. On the contrary, the predicted hospital mortality derived from the prognostic function tends to increase in 1980s. It is, therefore, concluded that the recent decline in hospital mortality of acute myocardial infarction observed is the result of progress and improvement in medical care. The way through which mortality was decreased was discussed. PMID- 8222978 TI - [A clinical study of intravenous captopril in heart failure]. AB - A clinical study of intravenous captopril was performed in 10 patients with mild heart failure and 20 with severe heart failure. The results indicated that intravenous captopril may rapidly reduce cardiac preload and afterload, increase cardiac output, inhibit renin system activity and decrease plasma level of catecholamine. A short period of treatment with intravenous captopril may produce a rapid symptomatic improvement in patients with acute or severe heart failure. There was a good tolerance with the therapy. PMID- 8222979 TI - [A clinical study on determination of plasma vasoactive intestinal peptide and the relationship between plasma vasoactive intestinal peptide and bronchial responsiveness in asthmatics]. AB - Vasoactive intestinal peptide (VIP), which is localized in normal human lung, may play an important role in regulating bronchial tone, pulmonary blood flow and mucus secretion. The level of plasma VIP and bronchial responsiveness were studied in patients with asthma, chronic bronchitis and the healthy subjects. The results showed that the level of plasma VIP in asthmatic patients during acute attack and symptom-free period was significantly lower than that in the patients with bronchitis and the healthy subjects and it is negatively correlated with the bronchial hyperresponsiveness. It is suggested that both asthmatic attack and bronchial tone are related with the decrease of VIP. PMID- 8222981 TI - [Diagnosis of Helicobacter pylori infection by 13C-urea breath test]. AB - Depending on the character for the Helicobacter pylori (HP) to decompose the urea by its endogenous urease, we have investigated the possibility to diagnose HP infection by 13C-urea breath test in 55 cases. Also, we compared the diagnostic results from 13C-urea breath test with those from culture, histology, and rapid urease test. The satisfied accuracy for the detection of HP infection with 13C urea breath test was indicated. PMID- 8222980 TI - [Clinical evaluation of HBV, HCV and HDV serum markers in patients with liver cirrhosis (LC) and hepatocellular carcinoma (HCC)]. AB - 11 kinds of HBV, HCV and HDV serum markers were investigated in 46 patients with HCC, 48 patients with LC, and 52 controls without liver disease. The prevalence of one out of HBV or HCV or HDV markers (M) in HCC and LC was 91.3% and 95.8% respectively, significantly higher than that in controls (17.3%). Positivity of HBV-M in HCC and LC was remarkably higher than those of HCV-M and HDV-M (P < 0.05). Prevalence of HCV-M in HBV-M negative HCC and LC was 66.7% and 75.0% respectively, significantly higher than that in HBV-M positive cases (P < 0.05). Frequency of viral replication in HCC and LC was significantly higher than that in controls (P < 0.01). The co-occurrence of two or three kinds of viral markers in HCC was more prevalent than that in LC (38% vs 14%, P < 0.05). Patients with coinfection from both HBV and HDV had a significantly younger age than those infected by HBV alone or infected by HCV (more than 10 years earlier). Among HCC and LC, 36% of HBV seronegative cases had HBV DNA detectable in their serum. Our data suggest that HCC and LC have a close association with the infection of HBV HCV and HDV especially HBV. Active viral replication and coinfection of several kinds of virus play on important role in the determination of HCC or LC development, and HDV appears to provide an additional risk for HCC and LC. In HBV M negative cases, HCV infection may be more important for HCC and LC development than HBV. PMID- 8222982 TI - [Incidence of NIDDM in Daqing and forecasting of NIDDM in China in 21st century]. AB - Based on the survey of the prevalence of NIDDM in 110 660 people aged 25-74 in 1986, the incidence of NIDDM was investigated in 1990 in 36 471 people (M/F 18,801:17,670) who had normal glucose tolerance and/or plasma glucose concentration less than 6.7 mmol/L 2-hour after breakfast (carbohydrate 80 g) four years ago in Daqing, Heilongjiang province, northeast China. 103 male and 88 female diabetics were diagnosed among the 36 471 subjects from June 1986 to June 1990. This result showed that the annual incidence of NIDDM in this area is 131/100,000 (137 in males, 125 in females), and is 130/100,000 standardized to the 1982's chinese population (95% confidence interval, 94-168/100,000). We may estimate that there would be 754 thousand new diabetics per year in 25-74 years old Chinese if the total population were 1.3 billion in China in the 21st century. PMID- 8222983 TI - [Aluminum in renal osteodystrophy]. AB - To evaluate the relationship between aluminum and renal osteodystrophy, iliac bone biopsies and bone aluminum staining were studied in 51 patients with end stage renal failure. In 27 of 51 cases were measured PTH, 1.25 (OH)2D3 levels and bone aluminum contents. We found that the incidence of renal osteodystrophy was 100%. The positive rate of bone aluminum staining was 12%. The rate was highest in low-turnover osteomalacia. Serum aluminum and bone aluminum contents were significantly higher than controls in three types of renal osteodystrophy on hemodialysis. Bone aluminum correlated with the duration of hemodialysis (r = 0.704), also with the serum calcium (r = 0.341). The duration of hemodialysis and bone aluminum contents were markedly longer and higher in positive group of bone aluminum staining than that of negative group (P < 0.001, respectively). Our data suggested that: (1) aluminum accumulation in bone was related to duration of hemodialysis, (2) aluminum may play important roles in the intervention of bone mineralization. PMID- 8222984 TI - [The advances in the research on gene therapy with transduced hepatocytes]. PMID- 8222985 TI - [The advances in the research on hepatocyte growth factor]. PMID- 8222986 TI - Soil quality in relation to soil pollution. AB - In comparison with the environmental compartments air and water, soil is an extremely complicated system, because properties of the soil and chemical, physical and biological characteristics vary greatly between different soil systems. This makes the development of general rules for quantitative evaluation of soil quality impossible. The large variation between the buffering capacities of various soils for different compounds also complicates the question. In The Netherlands, a system of reference values for various compounds has been developed, these reference values reflect situations in which there is a low degree of contamination. For heavy metals, these reference values consist of sliding scales incorporating the influence of a soil's clay and organic matter content. Examples for derivation of effect-based soil quality standards are briefly described. PMID- 8222987 TI - Arthropods as disease vectors in a changing environment. AB - Arthropod vectors need to acquire energy, moisture, hosts and shelter from their environment. Changing human populations and industrialization affect almost every aspect of the environment. In particular, the prospects of climatic warming, urbanization and vegetation changes have the potential to materially affect global patterns of vector-borne diseases. Global warming will enable the expansion of the geographical distributions of vectors. The population dynamics of vectors will change in response to extended seasons suitable for development followed by less severe winters. The incidence of epidemics is likely to change in response to an expected disproportionate increase in the frequency of extreme climatic events. The impact of such changes on each of the major vector-borne diseases is reviewed and projections are made on the likely global areas at risk from spread of disease vectors. Research needs are identified and response strategies are suggested in the context of the ever-increasing impact of human populations and industrial activity on the environment. PMID- 8222988 TI - Human tropical diseases in a changing environment. AB - A taxonomy of the human environment and of its components is set out as a basis for understanding the complex determinants of human diseases in the tropics. The scale, nature and trends of tropical diseases are related to changes in environmental components, life systems and habitats, and these are analysed, with emphasis on vector-borne diseases, especially malaria. Although it is feasible to predict the consequences of changing single climatic variables for specific parts of parasite and vector life cycles, the probable effects of simultaneously changing several variables are far more difficult to predict because many qualitative and quantitative determinants of disease are likely to be affected and the effects of covariance are poorly understood. Local and specific changes are likely to overshadow more global climatic changes. The effects of global temperature rise are explored, with particular emphasis on the basic case reproduction rate for two species of human malaria parasites. Even a 2 degrees C temperature rise greatly increases this rate at the lower end of the temperature range compatible with transmission. The importance of secondary vectors may increase. Implications for transmission in Europe and for imported tropical diseases are discussed. PMID- 8222989 TI - Human respiratory disease: environmental carcinogens and lung cancer risk. AB - Inhalatory intake of environmental agents may have adverse effects on health, the lung being the first target. Therefore, an increased risk of lung cancer and respiratory disease is in general considered as an indication of environmental health problems related to exposure to industrial emissions, traffic exhaust and smog. Classical epidemiological studies of the association between exposure to ambient air pollutants and respiratory dysfunctions and studies with laboratory animals have failed to demonstrate the distinct proof of risk for the general population that would be needed to form a basis for high impact environmental policy measures. Here, as an example, we describe the uncertainty in assessing risks of lung cancer associated with environmental exposure to polycyclic aromatic hydrocarbons. The recently introduced methodology of molecular cancer epidemiology is considered to yield more information on the relationship between exposure to environmental carcinogens and tumour development. Recent advances in the study of carcinogen (polycyclic aromatic hydrocarbon) dosimetry at the DNA level in combination with proto-oncogenic activation in humans are described. PMID- 8222990 TI - Radiation-induced disease. AB - The term radiation covers a wide spectrum of forms of energy, most of which have at one stage or another been suspected of causing human ill health. In general, study of the effects of radiation on health involves a mix of scientific disciplines, from population epidemiology to physics, which are seldom if ever found in a single scientist. As a result, interdisciplinary communication is of the utmost importance, and is a potent source of misunderstanding and misinformation. The forms of radiation which have been most specifically associated with health effects include ionizing and ultraviolet radiation. Claimed effects of electromagnetic and microwave radiation (excluding thermal effects) are too indefinite for detailed consideration. Ionizing radiation is a well-documented mutagen, which clearly causes cancers in humans, and human exposure has been increased by atomic weapons testing and medical and industrial uses of radioactivity. There is also a growing awareness of the possible role of some types of natural radiation, such as radon, in causing disease. Ultraviolet radiation is also associated with cancers, and is suspected of involvement in the increasing incidence of skin cancers in European populations. Factors thought to underlie recent changes in exposure to these mutagens are discussed. PMID- 8222991 TI - Reproductive health, population growth, economic development and environmental change. AB - World population will increase by 1000 million, or by 20%, within 10 years. Ninety-five per cent of this increase will occur in the South, in areas that are already economically, environmentally and politically fragile. Morbidity and mortality associated with reproduction will be greater in the current decade than in any period in human history. Annually, 40-60 million pregnancies will be terminated and 5-10 million children will die within one year of birth. AIDS related infections, e.g. tuberculosis, will undermine health care in Africa (and elsewhere) and in places AIDS-related deaths will decimate the work-force. The growth in population and associated morbidity will inhibit global economic development and spawn new problems. The key issues are migration, the spread of disease, the supply of water and the degradation of land, and fiscal policies with respect to family planning, pharmaceuticals and Third-World debt. Full education, particularly of women, and more effective family planning in the South have the power to unlock the problem. Failure will see the developed countries, with their 800 million population, swamped by the health, economic and environmental problems of the South, with its projected population of 5400 million people for the year 2000. PMID- 8222992 TI - CO2 and the greenhouse effect: present assessment and perspectives. AB - Our present knowledge on the increasing greenhouse effect is based on the 1990 assessment of the Intergovernmental Panel on Climate Change (IPCC) and its 1992 supplement. Model predictions suggest that a doubling of atmospheric CO2 concentration would increase global temperature by 2-4 degrees C. Because time is needed for the upper layers of the oceans to warm up, there is a delay between the realized increase and the estimated equilibrium increase. The 0.5 degrees C increase in global temperature over the past 100 years is in accordance with model predictions but also within the range of natural variability. The IPCC's assessment suggests that if fossil fuel consumption continues according to a 'business as usual' scenario, global temperature will increase at 0.3 degrees C per decade; droughts, flooding and storms would become more frequent and more severe, and sea-level would continue to rise. Analysis of gas in ice cores from Antarctica and Greenland provides estimates of CO2 concentrations in pre industrial ages; accurate measurement of atmospheric CO2 began in 1958. Atmospheric CO2 concentration has increased from 280 p.p.m. in AD 1800 to 355 p.p.m. at present. Between 1945 and 1973 global emissions of CO2 increased at a rate of 4.4% per year; after 1973 the rate of increase decreased to 1.6% per year. This change permits a test of the CO2 model. Reconstructed CO2 emission agrees within less than 10% with estimated fossil fuel-generated CO2 emission; contributions to CO2 emission from non-fossil fuel sources must be smaller than assumed previously. There are strong indications that in the past changes in atmospheric greenhouse gas concentrations inherent to the glacial-interglacial cycles did play an important climatic role. For example, they were probably responsible for interhemispheric coupling during the major climatic changes. Measures which might stabilize the greenhouse effect include energy conservation and improved energy efficiency, a transition to hydrogen rather than carbon as a source of fuel, and reforestation. PMID- 8222993 TI - Global epidemiology. AB - Epidemiology is the study of the distribution and determinants of health and disease in human populations. Epidemiology on a global scale is severely constrained by the lack of data. In many countries, there are no comprehensive data on mortality or basic demographic data. Where data are available, findings on the relationship of environment to health across countries need to be interpreted with caution. For example, there is well-known variation in standards of medical practice and diagnosis, and in certification and coding, but there are also large differences in diet, the social environment and lifestyle--all of which strongly predict disease incidence. Inappropriate inference concerning aetiology made from such broad-scale studies may result in what has been termed the 'ecological fallacy'. A complementary approach is to collect and analyse data in standardized fashion as part of international collaborative studies. These can offer some important advantages over the more conventional single-centre design. Recent advances have meant that studies of environment and health can now--in some countries--be carried out using routine data at the small area level. Although problems of interpretation remain, they are generally less severe than in broad-scale studies. Examples of this approach are given. PMID- 8222994 TI - Chemical pollution of the environment: past, present and future. AB - In an era of 'global environmental change' people are concerned about emissions of CO2, CH4, N2O and chlorofluorocarbons (CFCs) to the atmosphere because of their direct impact on global warming and their stratospheric ozone-depleting effects. Unprecedented efforts have been made to reduce the global emissions of CFCs. Major industries, which are competing within the modern global economy, have recognized the importance of maintaining a 'green' perspective. Future operations will be designed to reduce the direct emissions of chemical by products to air, water and soil, and to recycle and to reuse critical solvents such as water. 'Star Wars' technologies for the rapid, economical and effective elimination of industrial and domestic wastes will be developed and employed on a large scale. Advanced technologies for the control and monitoring of chemical pollutants on regional and global scales will be developed and implemented. Satellite-based instruments will be able to detect, to quantify, and to monitor a wide range of chemical pollutants. Our understanding of the fate and consequences of chemicals in the environment will increase dramatically such that we shall be able to predict the environmental, ecological and biochemical consequences of novel synthetic molecules with much greater precision. PMID- 8222995 TI - Environmental and health problems of developing countries. AB - Environmental variables largely determine the disease pattern in developing countries. Infections and malnutrition predominate, due to the effects of both poverty (a summary of many aspects of material deprivation) and a high ambient temperature. Environmental changes may be intended to improve health--examples include improved domestic water supplies and sanitation--or they may be due to socio-economic developments, which often have favourable or unfavourable health consequences. These are explored for water resource developments, where the health effects are complex, and for deforestation and urbanization. Although environmental impact assessment has been of value in reducing the adverse health impacts of socio-economic development projects, the use of health opportunity assessment is proposed as a more positive approach to optimizing the consequences of development to human health. PMID- 8222996 TI - Evolution and environmental change. AB - The effects of agriculture have been to improve the stock by selective breeding at a necessary loss of some capacities to withstand epidemics, while movements of products and stock have increased the speed with which epidemics spread. Some implications are considered. In relation to human genetics there is also evidence of a lack of selection, rather than any directed selection, and this will necessarily have some long-term effects. However, they are not well understood and are certainly slight, but cumulative. All that can be done at present is to maintain the lowest practical exposure to mutagens: the best understood, radioactivity, does not appear to be a serious hazard in the absence of catastrophic events or inappropriate medical usage. PMID- 8222997 TI - River Rhine: from sewer to the spring of life. AB - Water is the key issue in a number of declarations made by several eminent international commissions in recent years. The availability of clean and unpolluted water is crucial to sustainable development. The River Rhine was turned by pollution into the sewer of Western Europe; environmental protection measures, changes in industrial production and consumers' behaviour and remedial measures have drastically improved the quality of Rhine water, which, besides being the main water-way in Europe, also serves as a source of drinking water for a large population and is used for recreational purposes. Small occasional accidents, major spills, very remote accidents, war activities, etc. threaten the full recovery of what is the socially and economically most important watercourse in Europe. The organizational and technological measures taken to protect the Rhine from pollution can serve as an example of how other major freshwater sources could be protected from contamination or how existing pollution could be remedied. PMID- 8222998 TI - Assessing the greenhouse effect in agriculture. AB - Evidence that concentrations of CO2 and trace gases in the atmosphere have increased is irrefutable. Whether or not these increased concentrations will lead to climate changes is still open to debate. Direct effects of increased CO2 concentrations on physiological processes and individual plants have been demonstrated and the consequences for crop growth and production under various circumstances are evaluated with simulation models. The consequences of CO2 enrichment are considerable under optimal growing conditions. However, the majority of crops are grown under sub-optimal conditions where the effects of changes in CO2 are often less. The same holds for the possible indirect effects of environmental changes such as temperature rise. Studies on individual plants under optimal conditions are therefore not sufficient for evaluating the effects at a farm, regional, national or supra-national level. Simulation studies help to bridge the gap between the various aggregation levels and provide a basis for various studies of policy options at various aggregation levels. PMID- 8222999 TI - [Pre-and postoperative indocyanine green retention rate as a marker of liver functional reserve in cirrhotic patients]. AB - Indocyanine green ICG retention rate was determined before and after in portohypertensive cirrhotic patients undergoing paracardial devascularization (PCDV) and side-to-side mesocaval shunt (MCs-ss). The relationship between ICG retention rate with serum albumin level and the incidence of ascites was investigated no significant change the rate was noted before and after. Operation in PCDV patients. Significant increase of the rate was observed after MCs-ss. There was a negative correlation between the rate and serum albumin level. Addition the incidence of ascites increased with the elevation of the rate, there was a significant difference in the rate in patients with or without ascites. PMID- 8223000 TI - [Arthroscopic microdiscectomy]. AB - Forty six (42 cases) intervertebral discs were resected by the authors by means of arthroscopic microdiscectomy. Postoperative follow-up period was from 3 months to 6 months. According to MacNabis classification the result of treatment was excellent in 30 cases, which overs 71.4% of all patients. Failure of treatment occurred in 12 cases, this is mainly because of improper selection of cases. The authors expressed that this method is especially indicated in patients with simple disc protrusion. PMID- 8223001 TI - [Laser cholelithotripsy through choledochofiberscope: report of 30 cases]. AB - Laser cholelithotripsy was performed in 30 patients. Stones crushed by the laser were completely removed in 29 patients (96.7%) without any complications. Animal experiments and clinical application showed that the procedure caused no damage to the bile duct. But when the duct was flushed by flow water at a speed of not less than 15 ml a minute, with the tip of quartz fiber of Nd-YAG laser attached directly to the stone, the beam was radiated with 50 to 60 watt power for 1 minute each time in an interval of 1-3 minutes. Cholelithotripsy is indicated for patients with large or impacted residual stones after cholelithotomy, which can not be removed by choledochofiberscope and retrieval forceps. It can also be used to incise cycloid bile duct stricture with 50 watt power. PMID- 8223002 TI - [Autologous pericardial patch angioplasty of Budd-Chiari syndrome]. AB - 21 patients with inferior vena cava (IVC) or hepatic vein (HV) stenosis were treated by autologous pericardial patch angioplasty. Postoperative recovery was uneventful and the patients were followed up for 4 to 51 months. Signs of ascites, edema or varices in the lower extremities, and hepatomegaly disappeared in 1 to 3 months after operation in all patients. Ascending varicosities of the truncal vein and esophageal varices disappeared in 5 and 16 of '8 patients in 1 to 3 months respectively. Hepatic dysfunction in 6 patients returned to normal after operation. B-mode ultrasonography showed good patency of the reconstructed IVC or HV in the 18 patients. PMID- 8223003 TI - [Clinical studies of intra-thoracic stomach function in patients after esophagectomy and reconstruction by whole stomach]. AB - We usually use the stomach to hung up into the chest and to the neck for esophago gastrostomy in the patients of esophageal cancer. We had studied the intrathoracic stomach function in patients after esophagectomy with isotope 99m Tc labelled 717-resin semisolid meal for scintigram. We measured the gastric emptying time (GET) and fund the GET1/2 was no difference between the preoperative group and contrast group (P > 0.05). The study indicated that GET1/2 was faster obviously in postoperative patients with pyloroplasty than without pyloroplasty (P < 0.01). It was proved that to perform pyloroplasty with esophagostomy should be used routinely for preventing the pylorospasm, dilatation of the intrathoracic stomach and gastroesophageal reflux. At the same time, we found fasting serum gastrin (FSG) was increased (P < 0.01) in patients after esophagectomy than before, but basal acid output (BAO) decreased. It indicated that vagotomy caused the BAO decreasing and PH increasing. There were some relations between high level of FSG and postoperative diarrhea. PMID- 8223004 TI - [Surgical treatment of trilogy of Fallot in 42 adults]. AB - From 1984 to 1989 corrective operation circulation was performed under extracorporeal in 42 adults with trilogy of Fallot (6.2% of total intracardial operations under direct vision in patients with congenital heart disease in our hospital). The obstruction was relieved by resection of the hypertrophial and sclerotic muscle trabeculae of the Rt. ventricular outflow tract and pulmonary valvotomy and patching with a piece of artificial blood vessel lined internally by auto-pericardium so that a bougie (more than 1.6cm) could be passed through. The Rt. ventricular pressure was reduced satisfactorily (PRV/PLV < 0.75). There was one operative death. PMID- 8223005 TI - [Diagnosis and treatment of extra-adrenal pheochromocytomas]. AB - Eleven cases of extraadrenal pheochromocytoma were diagnosed and treated. When using general image and endoscopic examination, most tumors of this kind were accurately located. "Stimulant symptoms of tumors" were also helpful in locating the tumors. "Symptoms of tumor's clear away" were helpful in determining whether the tumor has been radically resected or not. The difficulty in operation is related to the occurrence of the tumor. If the tumor occurs in the postperitoneal region, there is a risk for the operation. Adequate preparation before operation is the key to a successful operation. Enucleation is not advisable for this tumor. PMID- 8223006 TI - [Purulent ventriculitis treated by continuous perfusion with antibiotic fluid]. AB - 24 cases of purulent ventriculitis were treated by perfusion with antibiotic fluid were reported. Two tubes were mounted, one for continuous drip in and the other for drain out. The results showed that it is more effective than ordinary antibiotic injection and/or single tube drainage. The cure rate may be promoted if effective antibiotic were used early. PMID- 8223007 TI - [Posterior transsphincteric approach for the surgery of the rectum]. AB - The posterior transsphincteric approach was used in the surgery of the rectum for 18 patients. They included 7 patients with rectal cancer, 7 villous adenoma malignant change 4, 1 carcinoid, and 3 benign stenosis, parasitic granulation and rectocele. The method of the operation and its main poits were described in detail; the indications of the operation were discussed. The anastomosis was influenced by the length, blood supply and cleaning of the proximal colon. Anastomotic leakage and pelvic infection were the main surgical complications. PMID- 8223008 TI - [Intraspinal tumor in the higher cervical region]. AB - Twenty-four cases of higher cervical intraspinal tumor were treated from July 1966 to Dec. 1990. Of them 22 cases had various paralysis according to Frankel's grading: A, 5 cases; B, 1; C, 9; D, 7 and E, 2. Having neurological symptoms for average of 5.4 months, all cases were operated. Total tumor resection were performed in 16 cases, partial resection in 5, spinal posterior decompression in 2 and one case was only bilpsed owing to abruptly respiratory arrest. Two cases died after operation. The results of operation were obviously improved in 16 cases. The authors introduce the diagnosis and operative skill of this kind of tumor as well as the treatment of disorders of respiratory function in this paper. PMID- 8223009 TI - [Diagnosis and treatment of secondary bone cysts]. AB - The article reported 17 cases of confirmed or suspected secondary bone cyst originating from cystic degeneration of pre-existed tumor or tumor-like lesions. Twelve cases were diagnosed as solitary bone cysts pathologically after operation, but their clinical or radiological manifestations were not in keeping with the pathological findings. Among them aggressive or malignant tumors appeared in 6 cases during follow-up and no primary lesions could be found in other 6 cases. The macroscopic features of the lesion of other 5 cases were very similar to a solitary bone cyst, nevertheless, primary tumors were found histologically. The authors held that when dealing with a cystic lesion of bone, it was important, but sometimes difficult to differentiate a solitary bone cyst from a secondary one, and suggested that if the latter was suspected, injection therapy should not be used, but more radical procedures should be taken for treatment instead and the patient should be observed closely for a longer period. PMID- 8223010 TI - [Generation and characterization of monoclonal antibody GCF-5 to human giant cell tumor of bone]. AB - Hybrid cell line has been derived from fusion of mouse myeloma (Sp2/0-Ag14) cell and spleen cell from mouse which was immunized with cultured cells of giant cell tumor of bone (GCT). Hybrid cells produce monoclonal antibody against GCT associated antigen in the surface of the tumor cells. The monoclonal antibody was detected by immunoperoxidase staining and was designated by GCF-5. There were more than eighty per cent of tested GCT which showed this antigen. It had no cross reaction with many kinds of normal tissue, HLA antigen, alpha-Fetal Protein, Carcino-Embryonic Antigen and beta-Microglobulin. The subclass of the antibody is IgG1. The hybridoma has been cultured and stored under -196 degrees C for more than eight months and its secretion remains stable. PMID- 8223011 TI - [Diagnosis and treatment of primary malignant fibrous histiocytoma of bone]. AB - Eleven cases of primary malignant fibrous histiocytoma of bone treated surgically were reported. There were 7 males and 4 females, with age ranged from 29 to 50 years (mean age 35.3 Yrs). The sites of involvement were femur (3 cases), tibia (3), humerus (2), radius (1) temporal bone, (1) and maxilla (1). The prominent findings in these cases were pain and swelling associated with regional mass. Differential diagnosis should be made from giant cell tumor, fibrosarcoma, osteosarcoma reticulum cell sarcoma and osteomyelitis. We in order to improve survival rate, think the ablative surgery should be the therapy of choice with post operational adjuvant chemotherapy. The five-year survival was 44% in these cases. PMID- 8223012 TI - [The application of liquid nitrogen cryosurgery in adjuvant to surgical treatment of spinal tumor]. AB - Liquid nitrogen (LN) cryosurgery is more often used for the treatment of bone tumors of the extremities. Since 1985 after animal experimentation we began to use it for the surgical treatment of spinal tumors. The temperature of the tissues surrounding the spinal cord was controlled above -20 degrees C by means of thermocouples and somatosensory evoked potentials (SEP) was used to monitor the spinal cord function. Liquid nitrogen was used as an adjuvant treatment in the surgical treatment of spinal tumors, it could reduce operative hemorrhage and kill tumor cells. The function of spinal cord had not been damaged in all 11 patients after cryotherapy. There was infection in one case, it was cured by using antibiotics and by drainage. This study showed that LN freezing can reduce the amount of operative hemorrhage during resection of spinal tumors. Benign tumors can be thus removed easily and thoroughly. PMID- 8223013 TI - [Aggressive fibromatosis of bone]. AB - Twenty four cases of pathologically diagnosed aggressive fibromatosis of bone were reported. 17 patients were followed for at least 2 years. Among them 4 recurred and the histological findings of their lesional tissues obtained after recurrence remained benign; one case refused radical treatment and the second biopsy done at 3.5 years after the appearance of the tumor showed no malignant changes; but another case died of lung metastasis. The authors suggest that the criteria for final diagnosis of an aggressive fibromatosis of bone are as follows: Besides the pathological findings, the lesion does not recur or metastasize more than 2 years after operation or it does recur, but still shows benign features histologically. As for treatment, marginal or wide excision is the procedure of choice. PMID- 8223014 TI - [Testicular tissue homotransplantation of newborn rat: experimental study]. AB - 32 newborn Sprague-Dawley rats were used as donors and 32 castrated male out-bred mature rats as recipients. The pieces of testes tissue were implanted subcutaneously in the bilateral hind legs. Two month follow-up showed that the grafts survived and increased in size. Serum testosterone levels of the recipients were higher than those of the castrated. They peaked and were close to the levels of the mature rats even 45 days after the implantation. Serum FSH, and, LH levels in the recipients were lower than those of the castrated. The mechanism of the newborn testes as an immuno-privileged donor is discussed. PMID- 8223015 TI - [The effect of glutamine on cholestasis caused by total parenteral nutrition]. AB - Glutamine is a tissue specific nutrient but not contained in standard TPN solution. To evaluate the effect of L-glutamine on TPN-induced cholestasis, 40 female Wistar rats were divided randomly into control or experimental groups. The rats were further subgrouped according to the days of observation and all received isocaloric (1008kJ.kg-1 x d-1) and isonitrogenous (1.22g.kg-1 x d-1) TPN support. 1% L-glutamine was given to the experimental group. Serum total bilirubin, albumin, lipoprotein X, r-GT, AKP and TG were determined on the day before study and 6th or 10th day after TPN. Before sacrifice, liver biopsies were taken for both light and electronic microscopic examinations, and the specimen of small intestine token for histologic evaluation. The results showed that the addition of glutamine to TPN solution can remarkably decrease the incidence, or ameliorate TPN induced cholestasis, and prevent small intestinal mucosal atrophy. PMID- 8223016 TI - [Comparison of the efficacy between exposure and moist ointment method in major burns: a report of 55 cases]. AB - Exposure therapy was compared with moist ointment method in 55 patients suffering from middle and large sized burns. Invasive infection occurred in 5 cases among 29 patients who were managed with exposure method, a rate of 17.2%, and one of them died as a result of infection, constituting a mortality rate 3.4%. On the other hand, 11 out of 26 patients (42.3%) who were treated with moist ointment developed septicemia, and 3 patients died of the infection (11.5%). The difference of both rate of invasive infection and mortality was statistically significant between two groups. In order to control the invasive infection we terminated moist ointment therapy in 3 cases, and 2 of them survived. To our opinion, the moist ointment does not do well on third-degree burn, especially in children with large and middle sized burns because of the ensuing rampant infection. PMID- 8223017 TI - [Rapid ultracentrifugal isolation and imaging detection of cholesterol phospholipid vesicles]. AB - We have utilized ultracentrifugation of bile-metrizamide density gradients to isolate and to quantitate the cholesterol-phospholipid vesicles (CPV) and identified them by transmission electron microscopy and scanning electron microscopy metrizamide was dissolved in bile and continuous density gradients were performed. The distribution of biliary cholesterol, phospholipid and bile salt was studied. The distribution of biliary phospholipid is similar in different fraction of the gradients. Approximately 70% of total biliary cholesterol was concentrated in the lightest fraction of the gradients (density < 1.035g/ml) and 70% of bile salt was found in the heaviest fraction of the gradients (density > 1.035g/ml). CPV harvested in fractions with density < 1.035g/ml and their diameter was 70-90nm. When 15% Metrizamide was dissolved in bile and centrifuged (50,000rpm, 100min, 37 degrees C), two thirds of total biliary cholesterol was concentrated in the top 0.4ml of the 5ml centrifuge tube. A lot of CPV was found in this part of bile and their diameter was 85nm. The advantage of the procedure described in this study is simple, rapid and accurate for pathological and physiological studies of CPV. PMID- 8223018 TI - [Long-term results of surgical resection of early esophageal and cardiac carcinomas]. AB - 204 patients with early esophageal and cardiac carcinomas were treated surgically. The survival rate of 5, 10, 15, 20 and over 25 years was 92.6%, 71.6% 62.7%, 50.9% and 48.0% respectively. Follow-up for 10-26 years showed that 88 patients died (13 lost to follow-up). Recurrent esophageal carcinoma accounted for 29.5% (26 Patients) and carcinoma of the second organ 21.6% (19). In this group, 4 patients with early infiltration carcinoma died from extensive metastasis after operation and 88.4% early esophageal carcinoma showed multiple foci. We consider that the principles for radical resection should also be followed for early esophageal carcinoma. PMID- 8223019 TI - [Analysis of surgical therapeutic effects in 135 cases of early bronchogenic lung cancer]. AB - 135 cases of early bronchogenic carcinoma (T1N0M0) out of 864 operated and pathologically confirmed ones were reported, with an incidence of 15.6%. There were 24 females and 111 males. The ages ranged 23-83 with a mean value of 58.7. There was no operative death. Pathological classification showed squamous cell carcinoma 43.7%, adenocarcinoma 20.0%, bronchiolealveolar carcinoma 25.9%. The postoperative follow-up study covered a period of 1-21 years. The 5-year survival rate was 70.6%, 76.2% and 83.3% respectively. Among this series 42.2% were of central type and 57.8% peripheral type. Their 5-year survival rates were 64.7% and 83.0% respectively, with significant difference (P < 0.01). The clinical features, early diagnosis, and surgical therapeutic effects of early bronchogenic carcinoma and multiple primary lung cancer are discussed. PMID- 8223020 TI - [Sleeve (bronchoplastic) lobectomy for the treatment of carcinoma of the lung: report of 135 cases]. AB - 135 cases of lung cancer were treated by sleeve (bronchoplastic) lobectomy within 24 years. 90 (66.7%) of them were of squamous cell carcinoma. Postoperative TNM classification showed 81 cases (60.0%) of stage I disease, 33 (24.4%) of stage II, 20 (14.8%) of stage III and 1 (0.7%) of stage IV. Upper lobe sleeve resection was used in 77.7% of the cases (right upper 57, left upper 48). Left upper sleeve lobectomy in 5 cases was accompanied by sleeve resection and reanastomosis of the left main pulmonary artery. One patient (0.7%) died of pulmonary thrombosis. The morbidity rate was 6.6%. Late complications included scar stricture of the anastomosis in 16 cases and recurrence at the site of anastomosis in 5. The 5- and 10-year survival rate was 56.2% and 42.9% respectively. For squamous cell carcinoma in particular, the 5-year survival was 60%. The indications and techniques of this operation are discussed in detail. Sleeve lobectomy had the advantages of preserving as much normal lung tissue as possible while keeping the principle of radical resection uncompromised. The low operative mortality and good long-term survival warrant its popularization. PMID- 8223021 TI - [Evaluation of local resection for primary tracheal tumor: a clinical analysis of 23 cases]. AB - During a 17 year period, 23 patients with primary tracheal tumor underwent surgery in the PUMC Hospital. Of them, 11 cases were benign tumors and 12 had low malignancies. 11 patients with benign tumors received 14 times of apoxesis of the tumor. 12 cases with low malignancies underwent 15 times of operations including local resection of the tracheal wall and tumor in 4 cases, and tumor apoxesis as well as electrocautery on the basis in 10 cases. Eight of 9 patients with adenoid cystic carcinoma were treated postoperatively with irradiation. Eleven cases with benign tumors were followed up for an average 6.3 of years. Nine cases with adenoid cystic carcinoma were followed up for an average 7.6 of years and 5 year survival rate was 75% (6/8). Among them 3 cases survived longer than 10 years. The authors consider that the local resection and tumor apoxesis can yet be regarded as a simple and effective way of surgical treatment for primary tracheal tumor. PMID- 8223022 TI - [Clinical significance of serum SCC Ag in patients with squamous cell carcinoma of the lung]. AB - To evaluate its clinical significance as a tumor marker, the serum level of squamous cell carcinoma associated antigen (SCC Ag) was studied by immunoradiometric assay in 59 patients with histologically proved lung cancer (32 squamous cell carcinomas, 23 adenocarcinomas and 4 small cell carcinomas). Carcinoembryonic antigen (CEA) was measured by radioimmunoassay simultaneously. The serum levels of SCC Ag and CEA were found above the normal range in 71.9% and 28.1% of squamous cell carcinomas, 26.1% and 34.8% of adenocarcinomas and 0% and 25% of small cell carcinomas respectively. In squamous cell carcinomas, the positive rate of SCC Ag increased with progression of clinical stage. With regard to the preoperative serum level dropped dramatically after surgery (P < 0.05). The change would be more outstanding after radical surgery (P < 0.001). The serum level of SCC Ag can be used to effectively monitor the course of disease, and could be used as an important item during the postoperative follow-up. PMID- 8223023 TI - [Surgical management of huge mediastinal tumor: report of 8 cases]. AB - The experiences of surgical management of 8 cases of huge mediastinal tumors were reported. Five cases had anterior mediastinal tumors, 3 posterior mediastinal tumors (benign 5, malignant 3). Emergent operations were done in 3 cases of intratumor hemorrhage and rupture. The average weight of resectal specimens was 2,080g. The clinical specificity of huge mediastinal tumor and management of pulmonary edema after operation were discussed suggesting a diagnostic standard and three methods: rapid tumor resection in capsule; resection of the tumor in pieces by minimizing the pressure within the tumor; resection of the tumor after enlargement of operation field. PMID- 8223024 TI - [Effects of obstructive jaundice on immunocompetence in jaundiced patients]. AB - Using CD series monoclonal antibody indirect immune fluorescent assay and LDH enzyme-release assay, we investigated lymphocyte subsets and natural killer cells (NKC) activity in the peripheral blood of 30 patients with obstructive jaundice. The results showed that the percentage of CD3+, CD4+, B cells and that of CD8+ was high. The ratio of CD4/CD8 was low at the first 10 days of jaundice when the serum bilirubin level was less than 171 mumol/L. Our studies suggested that obstructive jaundice might cause depression and impairment of host cellular and humoral immunity. Further studies showed that impaired immune function of jaundiced patients returned to normal 30 days after the disappearance of the jaundice by external or internal biliary drainage. Immune function recovered 30 days after relief of biliary obstruction and external or internal biliary drainage. External or internal biliary drainage was useful in improving host immunity. PMID- 8223025 TI - [Microwave surgery in the treatment of hepatic cancer]. AB - Microwave surgery was used to treat 50 patients with primary liver cancer (PLC) (hepatectomy for 46 patients and in situ coagulation of tumor for 4 patients with unresectable PLC). For hepatectomy, the needle electrode was inserted into the liver parenchyma repeatedly to coagulate the liver tissue at 1 cm interval along the predetermined incision line. For unresectable PLC, the needle electrode was directly inserted into PLC to coagulate the cancer tissue in situ. The average amounts of blood loss and blood transfusion for 46 hepatectomies was 215 +/- 189ml and 274 +/- 261ml respectively. It was significantly different from that observed in 46 matched conventional hepatectomies (P < 0.01). There were no operative mortality and complications such as delayed bleeding, bile leakage, and intraabdominal infection. These results indicate that microwave surgery can be utilized safely and effectively in the field of liver surgery. PMID- 8223026 TI - [Surgical treatment of lymphangioma of the neck]. AB - We studied 54 surgically treated cases of lymphangioma of neck, including total resection in 41 cases and partial resection combined with cryosurgery in 13. No recurrence was found in all 52 followed up cases. Facial Paralysis after operation occurred in 10 cases, among them, 9 made a recovery during a period of 2 to 6 months. We suggest that partial resection is the procedure of choice in order not to damage important surrounding structure in those cases and that total resection is not feasible. This relapse-free long term survival. PMID- 8223027 TI - [Primary arterialization of the deep vein in the treatment of severely ischemic limbs]. AB - Revascularization of severely ischemic limbs was performed on 212 limbs of 156 patients by creating an arteriovenous fistula between the arterial trunk proximal to the obliteration and a deep venous trunk of the ischemic limb, constricting the venous trunk proximal to the anastomosis by two thirds of its original diameter, ligating rami communicans and small tributaries of the deep vein distal to the fistula. The results of experimental and clinical studies showed that the ischemic limb was quickly revascularized and the cardiac function was not damaged after the operation, and the result is more satisfactory than that of arteriovenous reversal by stages. PMID- 8223028 TI - [Anterior displacement of tibial tubercle in the treatment of patellofemoral arthritis]. AB - This paper reports 41 cases (44 knees) with Patellofemoral arthritis treated by anterior displacement of tibial tubercle (modification of the classic maquet procedure of anterior displacement of tibial tubercle). Their average age was 47.5 years. 31 cases (34 knees) were followed-up for an average of 5 years and 7 months. Excellent results amounted to 64% and good results to 26%. Among theme, 12 cases were malalignment, 1 with lateral displacement of patella, 1 with subdislocation of patella and 7 with varus deformity of knee. Anterior displacement of tibial tubercle plus simultaneous proximal realignment operation was performed in 3 cases, plus distal realignment operation in 9 cases and plus coventry operation in 7 cases. PMID- 8223029 TI - [Observation of pregnancy and labour after kidney transplantation]. AB - Three patients were pregnancy after kidney transplantation were observed. One patient had omen labour 34 weeks after pregnancy owing to change of kidney function and caesarean section was done. The transplanted kidney completely lost its function after operation and was removed. The second patient received caesarean section 35 weeks after pregnancy due to raising of blood pressure and proteinuria. The patient had rejection of transplanted kidney and general failure after operation, and died 7 months later. The third patient showed rejection of transplanted kidney 21 weeks after pregnancy and was subjected to hydrostatic dilatation. The transplanted kidney completely lost its function and was removed. PMID- 8223030 TI - [Detective efficacy of abnormality during mechanical ventilation]. AB - The sensitivity of expired oxygen concentration (FeO2), oxygen concentration difference between inspiration and expiration (FiO2-FeO2), end-tidal CO2 (ETCO2) and pulse oxymetry (SpO2) in monitoring of several abnormal states during mechanical ventilation were compared in 10 patients. FiO2-FeO2 and FeO2 were found as sensitive as ETCO2 in monitoring hypoventilation. Oxygen analyzer located at the expiratory limb provided more information than at the inspiratory one. SpO2 was a rather insensitive detector for hypoventilation or even interruption of ventilation if FiO2 values was higher than that in ambient air. ETCO2 was a valuable estimate of adequate ventilation and CO2 elimination. PMID- 8223031 TI - [Origin of bone hyperplasia in osteoarthritic femoral head: a scanning electron microscopic study]. AB - Total hip replacement was carried out in 7 Patients suffering from osteoarthritis of the hip joint. They were given tetracycline preoperatively to label the newly formed bone tissues. The excised femoral heads were processed into undecalcified specimens, which were subjected to fluorescence microscopic and scanning electron microscopic studies. Narrow ribbon-shaped golden fluorescence could be observed along the trabeculae, reflecting newly formed bone by the cambium layer of the periosteum, while in the intertrabecular space both reticular and particulate and ball-shaped fluorescence could be detected, implying newly deposited bone by the marrow stromal cells. Under scanning electron microscope, the reticular new bone produced the new secondary trabeculae. The particulate and ball-shaped new bone tissues were deposited on the surface of the trabecular periosteum and filled up the inter-trabecular space. All these new bone tissues accounted for the bone hyperplasia or the osteoarthritic femoral head. PMID- 8223032 TI - [Study of antibody against protein in human allografts of decalcified bones]. AB - The result of study on antibody against protein in human allograft of decalcified bones was reported in this paper. 50 cases with allografts of decalcified bones were detected for sero-antibodies by indirect method of ELISA. The antigens used were extracted protein from the human decalcified bones. Among them, 32 cases showed positive reaction including complete bone graft healing in 30 cases, incomplete healing in one and failed in one cases success rate 98% The other 18 cases showed negative reaction. Neither the Positivith nor the titer of sero antibody interfered with the healing of allograft. PMID- 8223033 TI - [Intertrochanteric osteotomy in the treatment of osteoarthritis of the hip]. PMID- 8223034 TI - [Humoral and cellular immune responses to retinal S-antigen in uveitis patients]. AB - Humoral and cellular immune responses to purified S-Ag were determined in 73 patients with anterior-, intermediate- and pan-uveitis and in 55 healthy subjects by ELISA and leucocyte migration inhibition test. The positive rates of humoral immune response to S-Ag in patients and controls were comparable; however, the positive rate of cellular immune response to S-Ag in the patients (54.6%) was significantly higher than that in the controls (3.3%), and that in pan-uveitis patients (69.4%) was significantly higher than in patients with anterior uveitis (15.4%), highly suggesting that the cellular immune response to S-Ag was involved in the onset of uveitis, especially when the choroid and retina were affected. The breakdown of anterior chamber associated immune deviation and abnormal expression of intraocular MHC-II antigens may contribute to the development of cellular immune response. More studies are needed. PMID- 8223035 TI - [Cellular immunoreaction to S-antigen in patients with uveitis and optic neuritis]. AB - In vitro lymphocyte stimulation reaction to retinal S-antigen was evaluated in 115 patients of anterior uveitis, posterior uveitis/pan-uveitis, and optic neuritis, the positive rate being 34% (18/53), 41.5% (17/41), and 57.1% (12/21) respectively. It is speculated that such immunoresponse may be secondary to the exposition or the release of retinal antigens by various causes, leading to activation or augmentation of meager or low-affinity S-antigen-specific lymphocytes in healthy individuals to start the pathogenic autoimmune process. However, the authors also found that the stimulation index to PHA differed insignificantly between the control and experimental groups, indicating that there was no cellular immuno-hypofunction in the experimental patients. PMID- 8223036 TI - [Peripheral lymphocyte subsets in patients with Vogt-Koyanagi-Harada syndrome (VKH)]. AB - The peripheral lymphocyte subsets were analyzed in 37 patients with VKH and 32 normal controls by indirect fluorescein assay, using monoclonal antibodies. The percentages of OKT3+, OKT4+, and OKT8+ T-cells in active patients were found to differ insignificantly from those in quiescent patients, as were those in patients of the VK type from those in patients of the Harada type (P > 0.05), whereas the percentage of OKT4+ was significantly higher, that of OKT8+ was lower, and the OKT4/OKT8 ratio was higher in the recurrent patients than in the controls (P < 0.01). There was statistical difference in the ratio of OKT4/OKT8 between the recurrent and the non-recurrent patients (P < 0.01), while the ratio differed insignificantly between the two types of recurrent patients. The results suggested that the recurrence of VKH could be related with imbalance between the T-cell subsets. PMID- 8223037 TI - [An immuno-pathological study of pterygium]. AB - Pterygium specimens from 68 patients and normal conjunctiva from 12 controls were examined for the contents of immunoglobulins and C3 by direct immunofluorescence assay. The levels of IgE, IgG and IgA in the epithelial layer, of IgE, IgG and C3 in the basement membrane, and of IgE, IgG in the stroma, were found significantly higher in progressive cases than in the controls. as was the IgG-bearing cell count in the stroma. Also IgG bound to the basement membrane was significantly manifest in the quiescent cases. Routine histologic staining showed an infiltration of lymphocytes and plasma cells in the pterygium. A small amount of immunoglobulins was found in the stroma from the controls, but no C3 was demonstrated. These results suggest that an immunologic mechanism, possibly type I and type III hypersensitivity, may contribute to the pathogenesis of pterygium. PMID- 8223038 TI - [Treatment of blepharospasm and hemifacial spasm with botulinum A toxin]. AB - 112 patients with blepharospasm or hemifacial spasm were satisfactorily treated by local injections of small doses of botulinum A toxin, resulting in rapid relief or alleviation of the symptoms. No systemic adverse reactions were noted, and local side effects were minor and transient. The study shows that the therapy is safe and effective as a simple and easy alternative of surgical intervention. PMID- 8223039 TI - [Efficacy of argon and Nd:YAG laser combined in the treatment of primary chronic closed-angle glaucoma]. AB - 22 cases (29 eyes) of primary chronic closed-angle glaucoma were treated with Argon laser gonioplasty (ALG) and Nd:YAG laser iridectomy. Among 8 eyes in which the closed-angle was not opened by ALG, Nd:YAG laser gonioplasty was added, and 6 eyes became successful. For 9 eyes in which the IOP failed to drop inspite of the opened anterior chamber angle, Argon laser trabeculoplasty was added to bring remarkable curative effects to 7 eyes. During a follow-up of 5 to 14 months, 25 eyes (86%) obtained satisfactory results. PMID- 8223040 TI - [Early diagnosis and treatment of contralateral primary glaucoma associated with neovascular glaucoma]. AB - The early diagnosis and treatment of contralateral primary glaucoma associated with neovascular glaucoma (NVG) are discussed. Treatment of the underlying disease of NVG and long term follow-up of both eyes are emphasized in order to prevent blindness in both eyes. PMID- 8223041 TI - [Clinical analysis of steroid glaucoma]. AB - During 1970-1990, the authors accumulated 15 cases (26 eyes) of steroid glaucoma due to topical administration of dexamethasone or prednisolone. The patients comprised 10 men and 5 women, averaging 28 years of age. The clinical course was like chronic open angle glaucoma in 25 eyes, and 1 eye simulated an acute glaucoma attack. On presentation, the IOP was over 6.7 kPa in 9 eyes despite of antiglaucoma medication. The C/D ratio was > 0.6 in 9 cases (16 eyes), and the values were inconsistent between both eyes in 55% of the patients. Treatment was immediate discontinuation of the steroids, with antiglaucoma medication if necessary. 3 eyes were done filtering operations and 2 eyes had the subconjunctival residual steroid injection removed. Normal IOP was restored in months up to a year. PMID- 8223042 TI - [Long-term curative effect of cryotherapy for malignant ocular tumours]. AB - 66 malignant cases (2.7%) among 2401 patients of ocular tumours were treated with cryotherapy, and the tumours disappeared with satisfactory functional and cosmetic results. The cases were followed up 3-10 years (70% over 5 years), averaging 6.5 years. 6 cases (9%) recurred, 5 cases in the first and 1 case in the 9th postoperative year. The author analysed the causes of recurrence and discussed means to prevent it. PMID- 8223043 TI - [Restoration of normal binocular vision after successful surgical correction of concomitant exotropia]. PMID- 8223044 TI - [Surgical treatment of A-pattern strabismus with over-acting superior oblique muscles]. AB - 13 cases of A-pattern strabismus with overacting superior oblique muscles were treated with Berke's procedure. Depending on the severity of superior oblique muscle overaction, simple tenotomy or partial tenectomy of the superior obliques was performed. For cases of A-pattern strabismus < 30 prism diopters, bilateral superior oblique muscle weakening might result in overcorrection, and for cases > 40 prism diopters, bilateral superior oblique weakening should be accompanied with vertical displacement of the horizontal muscles. For patients with stereoacuity of 40 arc seconds, surgery should be refrained from in order to avoid post-operative cyclodeviation. PMID- 8223045 TI - [Long-term follow up of dacryorhinostomy in 14 cases with fibroendoscopy]. AB - The orifice of anastomosis and the lower aperture of the nasolacrimal duct in 14 cases (16 eyes) after dacryorhinostomy were followed up with the fibroendoscope. It was found that the size of the office and positioning of the bony window were key factors for a successful operation. Postsurgical management was advised to continue for at least 3 months or preferably 6 months. Deviation of the nasal septum toward the affected side was not regarded as contraindication for the operation, and the authors recommended fibroendoscopy in the diagnosis and postsurgical examination of lacrimal passage diseases. PMID- 8223046 TI - [A study of spectral electroretinograms of color vision defects due to macular diseases]. AB - Studies of spectral electroretinograms in central serous chorioretinopathy (CSC) and hereditary macular degeneration (HMD) found that the b-wave amplitudes of CSC for most wavelengths were significantly decreased as compared with normal controls, and the b-wave amplitudes of HMD for most wavelengths were obviously lower than the normal values. Patients of HMD with type-A color vision defects (CVD) showed primarily a reduction of b-wave amplitudes for the full spectrum of 480-620 nm, and patients with type-I CVD a reduction of b-wave amplitudes for wavelengths of 480-560 nm. These differences might reflect differences in damages to the medium-and long-wave length cones. PMID- 8223047 TI - [The refractive state after posterior chamber IOL implantation]. AB - The spherical refraction in 100 eyes after extracapsular cataract extraction and IOL implantation was < +/- 3.00D in 99% of the cases and < +/- 2.00D in 90% of the cases. The corneal curvature of 42 eyes followed-up changed very significantly in 4 months postoperative. The spherical power in 35 cases followed up 3 months to over 1 year changed insignificantly, while astigmatism reduced significantly, and in 1/3 of the eyes the axis continued to change after 3 months. Therefore, the author was of the opinion that glasses could be fitted 3 months after the operation and refraction periodically rechecked for at least 1 year. PMID- 8223048 TI - [Observations and measurements of the valves of the orbital veins]. AB - The lumens of the superior ophthalmic vein, the intraorbital communicating branch of the angular vein and the supraorbital vein of 19 adult male cadavers (38 orbits) were studied to find lunate venous valves in 79% of the cadavers, or 68% of the orbits. Of the 71 valves found, 39% were in the communicating branch of the angular vein, 58% in the supraorbital vein, and 2.8% in the superior ophthalmic vein. The mean distances from the valves in the communicating branch of the angular vein and those in the supraorbital vein to their junction to the superior ophthalmic vein were respectively 2.12 +/- 1.46 mm and 3.05 +/- 1.84 mm. It was important to note that the valvular sinuses opened toward the cavernous sinus. PMID- 8223049 TI - [Effects of interleukin-2 on experimental uveitis and pinealitis]. AB - Interleukin-2 (IL-2) was injected intraperitoneally into Lewis rats immunized with interphotoreceptor retinoid-binding protein (IRBP) to study its effect on the development of experimental autoimmune uveoretinitis (EAU) and experimental autoimmune pinealitis (EAP) and on the immune response. It was found that the EAU and the EAP were significantly intensified by IL-2, and the peak of serum anti IRBP antibody appeared on the 7th day after immunization while that in the controls on the 26th day. These results, together with those previously documented that the development, severity and subsidence of EAU depended on the level of IL-2, tend to support the authors' hypothesis that excessively elevated IL-2 enhances the development and recurrence of human uveoretinitis. PMID- 8223050 TI - [Experimental study of corneal endothelial damage by Nd:YAG laser]. AB - Evaluation by scanning electron microscopy revealed that rabbit eyes that underwent experimental anterior capsulotomy by Q-switched Nd:YAG laser suffered no endothelial damage, while the endothelium was focally denuded in those that received laser shots focussed within 0.8 mm behind it, and the Descemet's membrane was disrupted by laser shots focussed at the endothelium. The implications of clinical laser application were discussed. PMID- 8223051 TI - Omission of temporary diversion in restorative proctocolectomy--is it safe? AB - PURPOSE: The aim of our study was to evaluate the safety and functional outcome of restorative proctocolectomy (RP) without diversion. METHODS: Fifty patients underwent RP without diversion for ulcerative colitis (82 percent), familial adenomatous polyposis (12 percent), and indeterminate colitis (6 percent). The perioperative course and functional outcome of these patients were compared with another group of 50 patients undergoing RP with diverting ileostomy during the same time period (1989-1991) and closely matched for age, gender, surgeon, diagnosis, extent and duration (median, 10 years) of colitis, prior colectomy (approximately 22 percent), steroid use (40 percent), type of pouch, distance of ileal pouch-anal anastomosis from the dentate line (median, 1.5 cm), and the duration of follow-up (median, 12 months). All patients had a stapled ileal pouch anal anastomosis without mucosectomy and a smooth conduct of the operation. RESULTS: There was no operative mortality. Anastomotic leaks and pelvic abscess were more common in patients without ileostomy (7/50 or 14 percent vs. 2/50 or 4 percent); 8 of these 9 patients were taking > or = 20 mg of prednisone/day. Septic complications requiring relaparotomy (6 percent vs. 0 percent), prolonged ileus, and fever of unknown origin (10 percent vs. 4 percent) were also more common in patients without ileostomy. Despite similar functional results at 6 weeks and at 12 months after initial pouch function, patients without ileostomy had a poorer quality of life index (5 vs. 8; 10 being best) in the early period (0-6 weeks) of pouch function. CONCLUSION: In equally favorable cases, RP without diversion is not as safe as RP with diversion, especially in patients taking > or = 20 mg of prednisone/day. PMID- 8223052 TI - Colpocystodefecography. AB - PURPOSE: To assess, in one single procedure, a complete study of the female pelvis, including the Douglas pouch. METHODS: Colpocystodefecography (CCD) combines vaginal opacification, voiding cystography, and defecography. Three hundred examinations are reviewed. RESULTS: Thanks to the simultaneous visualization of the pelvic structures, CCD proved to be more useful than clinical evaluation to diagnose prolapses and particularly Douglas pouch hernias (enteroceles), the clinical diagnosis of which was missed in 93 of 111 cases. Moreover, in addition to morphologic and functional information, CCD brings about a new insight in the study of pelvic organs reciprocal influences, should they be positive (supporting function) or negative (external compression). Finally, significative pelvic surgery and particularly hysterectomy enhances greatly the risk of enteroceles. CONCLUSION: CCD is helpful in the preoperative staging, especially in the selection of the surgical procedure that will least likely predispose to possible late postoperative complications such as vaginal prolapses or enteroceles. PMID- 8223053 TI - Attempts to aid the adaptation of pelvic pouch before temporary ileostomy closure. AB - Most patients experience a high stool frequency immediately following the closure of the temporary ileostomy after total colectomy and ileoanal pouch reconstruction. Adaptation occurs within the ensuing weeks to reach a plateau in about three months. Increasing volumes of liquid nutrients were injected, twice daily for two months, into the pelvic pouch through a mucous ileal fistula proximal to the pouch before closing the temporary ileostomy. With this method the number of evacuations per 24 hours was significantly reduced during the first few weeks following the reestablishment of intestinal continuity, compared with a control group (average, 8.5 vs. 18.2, respectively). Patients also had better continence and less urgency to defecate. We suggest this technique in patients undergoing pelvic ileal reconstruction with temporary ileostomy. PMID- 8223054 TI - Immunohistochemical detection of mutant P53 protein and human papillomavirus related E6 protein in anal cancers. AB - PURPOSE: The wild-type P53 protein, a product of the P53 gene, is a normal growth controlling protein. Mutation of the P53 gene generates a mutant P53 protein which promotes tumor formation through loss of growth suppression. Some of the agents responsible for P53 gene mutation are known, one of which may be tumorigenic human papillomavirus (HPV) infection. Anal cancers are demonstrating a changing trend in the affected population, from older females in the older reported series to younger males more recently. This may be a reflection of infection with tumorigenic HPV types 16 and 18. The E6 oncoprotein of these viruses inactivates the growth-controlling wild-type P53 protein. In this study, our purpose was to determine the incidence of mutant P53 and HPV-16 and 18 related E6 protein and their coexpression in anal cancers. METHODS: We examined 29 anal cancers immunohistochemically for mutant P53 protein, HPV 16 and 18 E6 protein, and coexpression of the two. RESULTS: Mutant P53 protein was present in 58.6 percent of anal cancers overall and in 85.7 percent of anal adenocarcinomas. E6 oncoprotein was present in five cases (17.2 percent), all of which were squamous-cell carcinomas. Coexpression of both mutant P53 and E6 proteins was seen in only three cases (10.3 percent). CONCLUSION: Although tumorigenic HPV may be an important cause for P53 gene mutation in anal cancers, perhaps other mutagenic factors play a predominant role. PMID- 8223055 TI - Dyschezia and rectocele--a marriage of convenience? Physiologic evaluation of the rectocele in a group of 52 women complaining of difficulty in evacuation. AB - Herniation of the anterior rectal wall into the lumen of the vagina (so called rectocele) may be encountered in patients who complain of constipation and emptying difficulties but it is difficult to ascertain whether this anatomic abnormality is an etiologic factor or a consequence of the dyschezia. PURPOSE: The aim of our study was to assess symptomatic, anatomic, and physiologic features encountered in women with a clearly defined rectocele in order to determine the predisposing factors, symptoms, functional associations, and effects on quantified rectal emptying. METHODS: Clinical, physiologic (manometry), and anatomic (evacuation proctography) assessments were carried out in 26 consecutive women (mean age, 47.6 +/- 12 years) with dyschezia and a large rectocele as evidenced by radiography and compared with a group of 26 consecutive women complaining of dyschezia without a significant rectocele (mean age, 42.6 +/ 14 years). Both groups were similar with respect to mean age, parity, laxative abuse, manual anal evacuation, fecal incontinence, urgency, and weekly stool frequency. RESULTS: Patients having a rectocele differed significantly from those without a rectocele in having frequent endovaginal digitation during defecation (7 vs. 1, P < 0.05), more frequent symptoms of urinary incontinence (14 vs. 3, P < 0.001), and a surgical history of hysterectomy (9 vs. 2, P < 0.05). The rectocele group differed in having a delayed rectal emptying (55.5 +/- 38 vs. 30.3 +/- 23 seconds, P < 0.005), a more frequent incomplete rectal emptying (23 vs. 11, P < 0.0005), and was more often associated with a manometric anismus (16 vs. 6, P < 0.01). During the straining effort, there was a correlation between the depth of the rectocele and the duration of rectal emptying (rs = 0.3, P < 0.05). In the group without manometric anismus, women with a rectocele (n = 10) had a more incomplete rectal emptying than those without rectocele (8/10 vs. 8/19, P = 0.05). CONCLUSION: Some of our results indicate that the rectocele itself could be a contributory factor in difficult evacuation. These results also exhibit the importance of other disorders, such as anismus, in the occurrence of dyschezia. Physiologic examination therefore should be made before considering surgical repair in any patient with rectocele and dyschezia. PMID- 8223056 TI - Sonographic, manometric, and myographic evaluation of the anal sphincters morphology and function. AB - PURPOSE: A direct comparison of anal sphincter physiology (muscle performance and anatomy-muscle thickness) has not yet been undertaken but may be of importance in patients with defecation disorders. METHODS: We evaluated 15 healthy volunteers by means of anorectal manometry to determine pressure functions of the internal and external and sphincter. Transcutaneous electromyography was recorded to assess the electrical activity of the external anal sphincter. Thickness of the anal sphincter muscles was measured sonographically from within the anal canal with dorsal projection using a 7.5-MHz 360 degrees rectal panorama scanner. RESULTS: It was shown that neither is the muscle thickness of the external anal sphincter during rest (6.26 +/- 1.02 mm) or during squeezing (7.40 +/- 1.39 mm) correlated to its squeeze pressure (138.8 +/- 15.2 mmHg), nor is the diameter of the internal anal sphincter (2.09 +/- 0.58 mm) correlated to its resting pressure (61.1 +/- 15.2 mmHg). Finally, the thicker the internal and sphincter muscle, the lower the electrical activity of the external anal sphincter during squeezing (r = 0.74, P < 0.001). CONCLUSION: It is concluded that both manometry to assess sphincter function and sonography to determine sphincter morphology are important measures in the evaluation of continence function. PMID- 8223057 TI - Treatment of advanced hemorrhoidal disease: a prospective, randomized comparison of cold scalpel vs. contact Nd:YAG laser. AB - PURPOSE: Recently, laser technology has been advocated for the treatment of hemorrhoids. However, there has been little scientific evaluation of the use of the Nd:YAG laser for excisional treatment of hemorrhoidal disease. The purpose of this study was to perform a prospective randomized study of the Nd:YAG laser vs. scalpel excision, when performing a standard Ferguson-closed hemorrhoidectomy. METHODS: Patients presenting for internal-external hemorrhoidectomy were eligible for study. Hemorrhoidectomies were performed under epidural or caudal blocks. The standard Ferguson closed hemorrhoidectomy technique was used. Data evaluated included: age, sex, estimated blood loss, operative time, postoperative pain scores, postoperative analgesic use, wound healing, and time for return to work. Eighty-six patients were eligible for study (laser, N = 51; scalpel, N = 35). RESULTS: There were no significant differences between the groups, except for a greater degree of wound inflammation and dehiscence at the 10 day postoperative visit for the laser group (laser, 1.7 +/- .2; scalpel, 0.8 +/- .2; P < 0.05, t test). The use of the Nd:YAG laser added $480 per case; as a result, the treatment cost for the laser group was $15,360 higher than that of the conventional group. CONCLUSION: The results indicate that there are no patient care advantages associated with the use of the Nd:YAG laser for excisional hemorrhoidectomy compared with scalpel excision. As new technology becomes available, surgeons must rigorously assess therapeutic efficacy and cost-benefit ratio before deciding to employ this technology for patient care. PMID- 8223058 TI - Is preoperative computerized tomography useful in assessing patients with colorectal carcinoma? AB - PURPOSE: Controversy exists over the appropriate preoperative evaluation of colorectal cancer patients. Most surgeons agree that basic laboratory studies are indicated. Computerized tomography of the abdomen and pelvis has been used in our practice to augment the preoperative evaluation of these patients. METHODS: One hundred fifty-eight consecutive patients with primary colorectal carcinoma underwent computerized tomography (CT) of the abdomen as part of their preoperative evaluation. Their medical records were retrospectively reviewed. RESULTS: In 88 patients, 120 findings present on CT were otherwise unknown. Of these, 35 percent were clinically significant in that they allowed the surgeon to alter the proposed operative procedure or added additional technical information for consideration preoperatively. Findings include liver metastasis (26), atrophic kidney (3), and abdominal wall or contiguous organ invasion (11). In addition, two other solid organ carcinomas were detected. In the remaining 70 patients, CT contributed no additional pertinent information about the patient prior to this initial operative procedure. CONCLUSIONS: CT aids in the preoperative evaluation of individuals with colorectal carcinoma. It provides important clinical information that is useful to the surgeon planning the procedure. Additionally, CT permits the patient and his family to be aware of their overall status and to subsequent treatment options. Computerized tomography eliminates the need for preoperative intravenous pyelogram, improves the preoperative staging for metastatic disease, and provides a baseline for comparison during the postoperative follow-up period should recurrence be suspected or adjuvant therapy be planned. PMID- 8223059 TI - Reticuloendothelial stimulation: levamisole compared. AB - PURPOSE: Levamisole in combination with 5-fluorouracil is an effective adjuvant for the treatment of resected Dukes stage C colon cancer. Since the mechanism of action of levamisole is not known, we have investigated its effects on hepatic and splenic reticuloendothelial system (RES) activity in the rat and compared the effect of levamisole with other known RES stimulators. METHODS: The hepatic and splenic uptake of an intravenous dose of technetium-99m-sulfur colloid has been used to measure RES activity in rats treated with levamisole, glucan, zymosan, chlormethiazole, octreotide, and saline. RESULTS: Levamisole significantly increased the hepatic uptake of technetium-99m-sulfur colloid and is comparable in its effect to the other RES stimulators. In contrast, levamisole has no effect on splenic RES activity. CONCLUSION: RES function is considered to be a potentially important factor in the development of liver metastases, and the stimulatory effect of levamisole on the hepatic RES may partly explain its efficacy as an adjuvant treatment in colon cancer. PMID- 8223060 TI - Life expectancy after colectomy and ileorectal anastomosis for familial adenomatous polyposis. AB - PURPOSE: Despite the introduction of screening, surveillance, and prophylactic colectomy surgery, patients with familial adenomatous polyposis (FAP) are at risk of dying from other malignancies. METHODS: In order to quantify this risk and identify the causes of mortality, a retrospective life table analysis was performed on 222 patients with familial adenomatous polyposis who had undergone a total colectomy and ileorectal anastomosis between 1948 and 1990. These FAP patients were compared with an age- and sex-matched group of the general population and a relative risk of dying was calculated. RESULTS: Of 222 patients, 53 have died. In a matched group of the general population the expected number of deaths would be 15.8. The relative risk of dying is therefore 3.35. There has been no significant improvement with time and the relative risk is greatest for female patients. CONCLUSION: The three main causes of mortality are upper gastrointestinal malignancy, desmoid disease, and perioperative complications. Further research should therefore be aimed at prevention and improved treatment of these in order to improve survival. PMID- 8223061 TI - Usefulness of carcinoembryonic antigen monitoring despite normal preoperative values in node-positive colon cancer patients. AB - PURPOSE: The aim of our study was to determine to what extent serial carcinoembryonic antigen (CEA) monitoring is helpful in detecting colorectal cancer recurrence in patients if their preoperative serum CEA is normal. Additional major objectives of this study were to correlate CEA immunohistochemical features of the primary tumor with serum CEA levels at the time of tumor recurrence in node-positive colorectal cancer patients with low preoperative CEA values. METHODS: One hundred fourteen node-positive colorectal cancer patients with preoperative serum CEA levels of < 5.0 ng/ml undergoing clinically curative operations were studied. Primary tumors were evaluated for tissue CEA using the same monoclonal antibody as used for serum CEA determinations utilizing the avidin-biotin-peroxidase immunohistochemical technique. RESULTS: The exact preoperative serum CEA value did not correlate with tumor grade, immunohistochemical CEA intensity or pattern. In the 32 patients who developed recurrent cancer, the serum CEA at recurrence was greater than 5 ng/ml in 44 percent. All such patients had CEA present in their primary tumor. There was no correlation with the exact preoperative serum CEA, the intensity of the primary tissue CEA, or the localization of such CEA and subsequent serum elevation at recurrence. CONCLUSION: Serum CEA is a useful marker in the detection of recurrent colorectal cancer despite normal preoperative values. PMID- 8223062 TI - Hinged anoscope. AB - A hinged anoscope for insertion of rectal catheters is presented. Catheters attached to or integrally part of an external monitoring device or collecting system can be inserted through this endoscope. The endoscope can be removed without disconnecting the catheter from the external device. PMID- 8223063 TI - Epidemiology of cancer of the colon and rectum. 1971. PMID- 8223064 TI - Prevention of hemorrhoids by controlled defecation. PMID- 8223065 TI - Practice parameters for the treatment of rectal carcinoma. The American Society of Colon and Rectal Surgeons. PMID- 8223066 TI - Biofeedback training in disordered defecation. A critical review. AB - For 20 years now, biofeedback applications in the treatment of fecal incontinence and, more recently, chronic constipation in adults have proven that improvement of anorectal function can be achieved in a majority of patients. Despite this evidence, the definitive mode of action of biofeedback training remains obscure but may include improvement of both motor and sensory functions of the anorectum. In addition, behavioral modification of individual defecatory behavior in these patients may be effective as well. The long-term results of such treatment need to be tested in future research. PMID- 8223067 TI - Abnormalities of innervation of internal anal sphincter in fecal incontinence. AB - Physiological and histological studies have shown that the internal and sphincter is abnormal in idiopathic fecal incontinence. We have recently demonstrated that the in vitro contractile response of the internal anal sphincter to the sympathetic neurotransmitter noradrenaline is decreased in incontinence. In this study we have further defined this reduced sensitivity and provided more information about the intrinsic innervation in both the normal and the incontinent sphincter muscle. Muscle strips from 12 incontinent patients undergoing post and repair and from 11 controls undergoing rectal excision for low rectal carcinoma were studied. Responses to noradrenaline were recorded initially alone and then in the presence of phentolamine, an alpha-adrenoceptor blocker. In the presence of phentolamine, noradrenaline caused relaxation: there was no significant difference in the relaxation-response curves and the EC50 was the same in the two groups. These results demonstrate that the previously documented reduced sensitivity to noradrenaline is due to an altered sensitivity of the alpha-adrenoceptors. Electrical field stimulation produced relaxations in all muscle strips, but only in the controls was the magnitude of the relaxation significantly increased in the presence of phentolamine. This indicates that there is an alpha-adrenergic excitatory component of the response to electrical field stimulation of the intramural nerves, which was present in tissues from control patients but which was absent in tissues from patients with idiopathic fecal incontinence. PMID- 8223068 TI - Anorectal stenosis in patients with prolonged use of suppositories containing paracetamol and acetylsalicylic acid. AB - We report five cases in which anorectal stenosis was associated with chronic administration of suppositories containing paracetamol and acetylsalicylic acid. All patients were females taking suppositories for chronic migraine. Patients experienced anal pain, tenesmus, fecal incontinence, and, in two cases, intestinal obstruction. Lesions were characterized by a severe circular narrowing of the distal rectum with superficial ulcerations. Radiographs demonstrated the presence of sinus-tract-like formations in the diseased segment and thickening of the rectal wall. Histologic examination showed signs of chronic inflammation with deep ulcerations associated with obliteration of the lamina propria by a fibromuscular proliferation, as was described in the solitary ulcer syndrome of the rectum. Rectal administration of suppositories was discontinued in all the patients. Two patients were improved by anorectal dilatation, but the remaining three required a left proctocolectomy with subsequent coloanal anastomosis. The use of such suppositories must be restricted. PMID- 8223069 TI - Colonic mucosal proliferation after pancreaticobiliary diversion in the hamster. AB - The effect of pancreaticobiliary diversion (PBD) on the colonic mucosa was studied in hamsters over 5, 10, and 24 days. Sham-operated animals served as controls. At all three time intervals, experimental animals had increased plasma cholecystokinin concentrations and decreased gastrin concentrations. Five days after PBD, there was an increase in scintigraphically measured [3H]thymidine incorporation into colonic tissue. Correspondingly, there was an increase in the [3H]thymidine DNA labeling index of goblet cells in the colonic mucosa. The total number of cells in the colonic crypt columns were significantly increased on days 5, 10 and 24. Whether this proliferative response in the colon is due to increased release of cholecystokinin, enteroglucagon, other aberrations of hormones or growth factors, or simply an increased bile load on the colonic mucosa remains to be clarified. Such further studies may reveal an alternative animal model for studies on risk factors in colonic carcinogenesis. PMID- 8223070 TI - Extraperitoneal manifestation of perforated diverticulitis. AB - We observed five cases of unusual extraperitoneal presentation of diverticulitis during an 11 year period. During that time, a total of 263 patients were operated for diverticulitis. Patients 1-4 presented with an inflammatory spread of diverticulitis through the abdominal wall; in patient 5 multiple abscesses were present in the left thigh. Inadequate resection and previous radiotherapy in patient 4 might have contributed to the complicated course. In patient 5, poor condition initially allowed only local drainage of the thigh abscesses. Despite laparotomy and sigmoid resection a few days after drainage, the fatal outcome could not be prevented. A more extensive resection and a protecting colostomy might have prevented the fistulous complication in patient 4. A primary resection and abscess drainage might have averted the fatal outcome in patient 5. PMID- 8223071 TI - Very low intraluminal colonic pH in patients with active ulcerative colitis. AB - Intraluminal gastrointestinal pH was measured in seven patients with active ulcerative colitis (four male, three female). A radiotelemetry capsule was used, and its location was determined by fluoroscopy. Satisfactory measurements were obtained from six, in all of whom pH levels were normal in the stomach and small intestine. Three patients also had normal pH values in the colon. However, in the remaining three patients very low pH levels (lowest values 2.3, 2.9, and 3.4) were found in the proximal parts of the colon. Five of the seven patients, including the three with low pH in the colon, underwent colectomy. The mechanism behind the low intraluminal pH in some patients with ulcerative colitis is speculative. Increased fecal concentrations of lactate occur in active disease, but some of the pH values measured in our study were below the pKa value of lactate. The study demonstrates that very low intraluminal pH levels in the colon occur in some patients with active ulcerative colitis. This might be an indicator of severe activity of the disease. PMID- 8223072 TI - Stress proteins in colorectal mucosa. Enhanced expression in ulcerative colitis. AB - Stress (heat shock) proteins are ubiquitous intracellular proteins that can be induced in vitro by physiological stress events that occur during inflammation. We have used an indirect immunoperoxidase method to locate 60-kDa stress proteins in biopsies taken from normal and inflamed colorectal mucosa. An anti-60-kDa monoclonal antibody (ML30) produced specific staining of surface epithelial cells localized to the site of the Golgi apparatus. In ulcerative colitis, there was an increased concentration of this stress protein compared with controls (P < or = 0.002) and also with a small group of active Crohn's colitis (P < or = 0.01), but no relationship between its concentration and disease activity. All biopsies also showed staining of goblet cells by ML30, suggesting a possible cross-reaction with mucin; electroblotting of crude but not purified mucin showed a faint 60-kDa band with ML30. We conclude that the 60-kDa stress protein is present in normal colorectal epithelial cells and is markedly induced in vivo in ulcerative colitis. Further, we suggest that since the 60-kDa protein functions as a molecular chaperone, it may associate with colonic mucin aiding in its synthesis and/or secretion. PMID- 8223073 TI - Reduced metabolic efficiency in patients with Crohn's disease. AB - Malnutrition is frequently seen in patients with inflammatory bowel disease, and parenteral or enteral nutrition is considered essential in this patient group. However, many patients with Crohn's disease have difficulties in gaining weight in response to overfeeding, suggesting reduced energy retention. Substrate utilization and nutrient balances as well as changes in body composition were followed in 10 patients with Crohn's disease immediately in the course of remission on low-dose steroid treatment, during an eight-day period of continuous enteral nutrition at constant (protocol 1:1.5-fold basal energy expenditure) and increasing (protocol 2:0.5- to 2.0-fold basal energy expenditure) nutrient supply. Energy, substrate, and nitrogen balances all became positive in response to overfeeding. However, fat was predominantly oxidized at an infusion rate of 1.2 g/kg body wt/day, whereas carbohydrates and proteins were effectively stored. A positive energy balance was reached at an energy infusion rate exceeding 31 kcal/kg body wt/day and corresponding substrate supplies of 1.6, 1.7, and 1.1 g/kg body wt/day for carbohydrates, fat, and protein, respectively. Nitrogen balance normalized at a supply of 0.14 g/kg body wt/day, which also reduced myofibrillar protein breakdown. Considering the relative contributions made by these nutrients in the diets, an accumulation of carbohydrates and protein but a depletion in fat became evident from nutrient balances. In fact, body weight increased by 0.12 kg/day, which was explained by an increased extracellular (+0.18 kg/day) and body cell mass (+0.04 kg/day) at reduced fat mass (-0.10 kg/day). Concomitantly, plasma T3 and insulin secretion both increased, whereas sympathetic nervous system activity decreased with overfeeding. This is contrary to data observed in healthy subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223074 TI - Prevalence and consistency of low breath H2 excretion following lactulose ingestion. Possible implications for the clinical use of the H2 breath test. AB - The clinical use of the H2 breath test is limited by the finding that a variable fraction of the population fails to excrete appreciable H2 during colonic carbohydrate fermentation. Therefore, we assessed the ability to increase breath H2 excretion in 371 patients (224 female, 147 male) by administering the nonabsorbable sugar lactulose. Following 12g of lactulose, 27% of 94 patients did not increase their breath H2 concentration over 20 ppm and were considered low H2 excretors. Ingestion of 20 g of lactulose in 277 patients yielded a frequency of low H2 excretors of 14%. Six of 10 patients that were low H2 excretors after 12g of lactulose increased their breath H2 levels over 20 ppm when tested with 20g. In 35 patients tested with the same amount of lactulose on two separate occasions, the subject frequently altered his or her H2 producing status over a period of a few weeks. Low H2 excretors had a significantly higher breath CH4 concentration, both fasting (22 +/- 34 ppm) and after lactulose (51 +/- 58 ppm) compared to the remaining patients (5 +/- 13 ppm and 16 +/- 40 ppm, respectively). While the mean age of low excretors (54 +/- 17 years) was significantly higher than the others (44 +/- 17 years), no difference was found for sex prevalence and stool pH. This study demonstrates that respiratory H2 excretion following lactulose ingestion is not consistent and suggests that the application of too restrictive criteria could lead to improper interpretation of the H2 breath test. PMID- 8223075 TI - Effect of long-term colchicine therapy on jejunal mucosa. AB - Colchicine is recommended as daily prophylactic therapy in patients with familial Mediterranean fever (FMF) to prevent febrile paroxysms. The drug is known to be a potent inhibitor of mitotic activity and might therefore be expected to have a significant adverse effect on tissues that undergo rapid turnover. We studied small bowel biopsies from nine patients with FMF who were receiving daily low dose oral colchicine therapy. In each patient the lengths of 20 crypts and villi were measured and the number of mitotic figures in 20 crypts were counted. The data were compared with similar measurements from histologically normal-appearing biopsies obtained from 14 patients with a variety of mild gastrointestinal complaints. The mean crypt length was found to be significantly greater (0.197 mm vs 0.186 mm, P < 0.0001) and the mean villous length significantly smaller (0.369 mm vs. 0.442 mm, P < 0.0001) in the FMF patients than in the control population. In addition, the mean number of mitotic figures per crypt was significantly higher in the FMF patients (2.58 vs 1.00, P < 0.001). The data reveal a pattern of mucosal injury in the colchicine-treated FMF patients characterized by a hyperplastic crypt-villous atrophy pattern with increased mitotic rate, which is indicative of an increase in cell turnover and opposite to what we anticipated based on colchicine's known effect on mitotic activity. PMID- 8223076 TI - Comparative effects of exogenous lactase (beta-galactosidase) preparations on in vivo lactose digestion. AB - Microbial-derived beta-galactosidase (beta-gal) enzyme preparations improve in vivo lactose digestion and tolerance through enhanced gastrointestinal digestion of lactose. Three different beta-gal preparations, Lactogest (soft gel capsule), Lactaid (caplet), and DairyEase (chewable tablet) and placebo were fed to lactose maldigesters with either 20 g or 50 g of lactose to compare the efficacy of these products and to further establish a dose-response relationship for use. All enzyme preparations dramatically reduced both the peak and total breath hydrogen production when fed with milk containing 20 g of lactose. Four capsules of Lactogest, two caplets of Lactaid, or two tablets of DairyEase (each treatment containing approx 6000 IU) reduced total hydrogen production significantly (P < 0.05) below that observed with two capsules of Lactogest (containing approx 3000 IU) in a stoichiometric manner. Symptoms were significantly (P < 0.05) less severe with all the beta-gal products. In contrast, with 50 g of lactose in water, peak and total hydrogen production was modestly, but not significantly reduced by the enzyme treatment. Furthermore, symptom scores for bloating, cramping, nausea, pain, diarrhea, and flatus were not different between treatments and the control. The 50-g lactose dose appeared to overwhelm the ability of either 3000 or 6000 IU of beta-gal to assist significantly with lactose digestion. Results from these studies demonstrate the relative equivalency of chewable, caplet, and soft-gel beta-gal products, based on IUs of enzyme fed. PMID- 8223077 TI - Value of endoscopic markers in celiac disease. AB - Duodenoscopy in celiac disease has identified several markers of the disease. Our aim was to evaluate, in a prospective study, the usefulness of the different endoscopic features in 100 consecutive cases referred to endoscopy for intestinal biopsy. Histological examination of duodenal samples showed severe villous atrophy (grade III/IV) in 36 patients. Of these patients, 34 had endoscopic markers suggestive of celiac disease. These were reduction in number or loss of Kerkring's folds (in 27), mosaic pattern (14), scalloped folds (12), and visibility of the underlying blood vessels (5). Endoscopic visualization of these markers had a sensitivity of 94%, a specificity of 92%, and a positive predictive value of 84%. Reduction in number, or loss of, Kerkring's folds was the most sensitive (76%) and specific (98%) single endoscopic change indicating celiac disease. Duodenoscopy permitted diagnosis in three of four asymptomatic patients in a group of 24 first-degree relatives of celiac disease patients. We conclude that endoscopy of distal duodenum is a sensitive and specific indicator of celiac disease. PMID- 8223078 TI - Serum IgA anti-gliadin antibodies in an adult population sample. High prevalence without celiac disease. AB - IgA-class anti-gliadin antibodies (AGA) and IgA-, IgG-, IgM-class anti-reticulin antibodies (ARA) were determined in 1461 persons, representing 84% of a population from the village of Karksi-Nuia. AGA were detected by enzyme-linked immunosorbent assay (ELISA) and ARA by indirect immunofluorescence. Fifty-two (3.5%) persons had IgA-class AGA, of whom 48 and an additional three of four persons with diarrhea were biopsied. All biopsies showed normal small intestinal mucosal architecture. All 1461 persons were negative for ARA. Our results demonstrate that AGA are frequently detected in an adult Estonian population and positivity increases with age in persons with normal small intestinal mucosa. Positivity for AGA does not predict silent undetected celiac disease but rather represents a normal response to dietary antigens in the elderly. Inability to detect ARA suggests that celiac disease does not exist in this population. As none of the AGA-positive but ARA-negative biopsied persons had celiac disease, ARA might be a more specific serologic marker for celiac disease than AGA. PMID- 8223079 TI - Gastric mucosal damage induced by nonsalicylate nonsteroidal antiinflammatory drugs in rats is mediated systemically. AB - The gastric toxicities of an enteric-coated formulation and conventional indomethacin were compared in rats. Both formulations were equally damaging to the mucosa, suggesting that topical damage was not the major route of injury. The importance of systemically mediated damage was further determined by gastrotoxicity dose-response curves and pyloric ligation experiments in which indomethacin was administered either orally or parenterally, or into stomach or duodenum with the pylorus occluded. Gastric damage was significantly higher in those groups that had received the drug parenterally or intraduodenally. The extent of deeper mucosal damage, assessed histologically, was greater in parenterally dosed rats. In further experiments, oral and parenteral routes of administration of two other nonsalicylate NSAIDs, naproxen and sodium diclofenac, were found to be equally damaging to the mucosa. Our results show that indomethacin-induced gastric damage, unlike aspirin injury, is mediated mainly systemically. Enteric-coating may not be a useful strategy in reducing gastric injury by nonsalicylate, nonsteroidal antiinflammatory drugs. PMID- 8223080 TI - Prostaglandins and sulfhydryls may mediate gastric protection induced by verapamil in rats. AB - Verapamil, a type-1 calcium-channel blocker, given intraperitoneally, macroscopically protected the gastric mucosa of rats from 96% ethanol-induced lesions in a dose-dependent fashion. This effect was significant when verapamil at 10 or 20 mg/kg was given 1 hr before ethanol. Histopathologically, verapamil prevented the development of deep necrotic lesions, but did not preserve the surface epithelium. Gastric acid secretion in both pylorus-ligated rats and gastric-diversion rats was inhibited by 20 mg/kg of verapamil. Gastric motility measured by a balloon method was dose-dependently inhibited by verapamil. Verapamil protection was significantly diminished by pretreatment with subcutaneous indomethacin (30 mg/kg) and iodoacetamide (100 mg/kg). The gastric motility inhibited by verapamil was not reversed by indomethacin and iodoacetamide. These results indicate the participation of both endogenous prostaglandins and sulfhydryls of the gastric mucosa in verapamil protection against ethanol damage, but do not relate to a suppression of gastric motility. PMID- 8223081 TI - Nonsteroidal antiinflammatory drugs and upper gastrointestinal hemorrhage in an urban hospital. AB - Aspirin and nonsteroidal antiinflammatory drugs have been implicated in the pathogenesis of gastrointestinal hemorrhage. To evaluate their impact on inpatients, charts from Temple University Hospital with a discharge ICD-9 code which included upper gastrointestinal hemorrhage during a one-year period were reviewed. Aspirin and/or nonaspirin nonsteroid antiinflammatory drug (NSAID) use was identified in 34 patients (19 daily users and 15 intermittent users). Sixty seven patients who bled, but did not use these agents, served as controls. Daily NSAID users were older than intermittent users and controls (P < 0.05). A higher frequency of bleeding ulcers was associated with NSAID use. Patients using NSAIDs spent more time in intensive care than controls (median 1 day vs 0 days). Daily users had a higher transfusion requirement (4 units) than non-users (0 units; P < 0.05). This study suggests that NSAID use has a substantial impact on health care resource utilization in patients admitted to an urban hospital for upper gastrointestinal hemorrhage. PMID- 8223082 TI - Prediction of therapeutic failure in patients with bleeding peptic ulcer treated with endoscopic injection. AB - Endoscopic injection therapy was performed in a consecutive series of 233 patients admitted for a bleeding peptic ulcer with active arterial hemorrhage or a nonbleeding visible vessel disclosed at emergency endoscopy. Further bleeding occurred in 57 cases (24.5%). The present study was conducted to evaluate whether any clinical or endoscopic features could identify the patients at high risk of therapeutic failure. Multiple logistic regression analysis showed that failure was significantly related to: (1) the ulcer location on the posterior wall (P = 0.004) or superior wall (P = 0.003) of the duodenal bulb, (2) the ulcer size (P = 0.011), and (3) the existence of associated diseases (P = 0.012). The validity of the prediction rule based on these factors was evaluated by receiver-operating characteristic curves and was confirmed and prospectively validated in an independent sample of 81 patients with a bleeding peptic ulcer treated by endoscopic injection. We conclude that once the initial control of bleeding has been achieved by injection therapy, the present prediction rule can be used to identify candidates for alternative treatment. PMID- 8223083 TI - Phospholipase activity of Helicobacter pylori and its inhibition by bismuth salts. Biochemical and biophysical studies. AB - In this study we measured phospholipase A (PLA) and C (PLC) activity of media filtrates and French Press lysates of the gastritis-inducing bacteria Helicobacter pylori. We report here that both H. pylori lysates and filtrates contain PLA1, PLA2, and C enzymes, which readily hydrolyze a radiolabeled dipalmitoylphosphatidylcholine (DPPC) and phosphorylcholine substrates, respectively. The specific activity of both PLA and C enzymes were greatest in the 6.5-7.0 and 8.4-8.8 pH ranges, respectively. Colloidal bismuth subcitrate (CBS) induced a dose-dependent inhibition of PLA2 and C activity of both H. pylori lysates and filtrates. This inhibitory effect of CBS on PLA2 was antagonized in a dose-dependent fashion by the addition of CaCl2 to the incubation mixture, suggesting that calcium and bismuth may be competing for the same site on the enzyme. In contrast, the ability of bismuth salts to inhibit PLC activity of H. pylori lysates was not antagonized by CaCl2. Employing a biophysical assay system for surface wettability, it was determined that H. pylori lysates had the capacity to remove a synthetic phospholipid monolayer off a glass in a dose-dependent fashion. This ability of the bacterial lysates to catalyze the transformation of a hydrophobic surface to a wettable state was significantly attenuated in the presence of bismuth salts. Our experimental results are, therefore, consistent with the possibility that H. pylori colonization compromises the stomach's barrier to acid by eroding a phospholipid lining, possibly a monolayer, on the surface of the gastric mucus gel and that this process is blocked in response to bismuth therapy. PMID- 8223084 TI - Serum pepsinogens as markers of response to therapy for Helicobacter pylori gastritis. AB - We have investigated the effect of therapy for Helicobacter pylori gastritis on serum concentrations of pepsinogen I and II in 43 patients. In the 22 patients in whom therapy resulted in dramatic decrease in gastritis scores and in clearance of the bacteria, there was a highly significant (P = 0.0001) fall in mean serum pepsinogen II from 13.3 +/- 0.8 to 7.9 +/- 0.7 micrograms/liter, and a less pronounced fall in pepsinogen I from 89.0 +/- 5.9 to 78.5 +/- 0.4 micrograms/liter (P = 0.01). These changes resulted in a significant (P = 0.01) increase in the pepsinogen I/II ratio. In contrast, nonsignificant declines of 3.5% and 11.6% were observed in mean pepsinogen I and II levels in the 21 patients whose gastritis failed to resolve histologically and whose infection did not clear. These findings suggest that serum pepsinogen levels, especially pepsinogen II, are a new tool that may be found to be clinically useful in evaluation of treatment outcome in patients with H. pylori-associated gastritis. PMID- 8223085 TI - Gastrospirillum hominis in asymptomatic, healthy individuals. AB - Gastrospirillum hominis is a spiral-shaped bacterium found in the stomach. It has been implicated as a possible cause of chronic gastritis. We report two cases of G. hominis colonization observed in a series of 175 healthy, asymptomatic volunteers investigated for Helicobacter pylori. None of the volunteers had symptoms or a history of gastrointestinal disease. Both carriers of G. hominis had histological signs of chronic, active antral gastritis. Multiple tests for H. pylori were negative. The prevalence of this spiral bacterium in healthy, asymptomatic individuals may be as low as in symptomatic persons. PMID- 8223086 TI - Influence of pancreatic ducts on saturation of juice with calcium carbonate in dogs. AB - In several species, bicarbonate and calcium concentrations of pancreatic juice are known to vary during the different phases of pancreatic secretion. The effects of these variations on the saturation of juice with calcium carbonate, a critical factor for the formation of pancreatic stones, are not known. In this work, we studied the saturation degree of pancreatic juice with calcium carbonate in six unanesthetized beagle dogs equipped with Thomas cannulae during basal secretion and after bolus injections of cerulein (30 ng/kg) or secretin (0.25 units/kg). In the different samples of pure pancreatic juice, pH, PCO2, bicarbonate, and proteins were measured by standard methods. Total calcium (CaT) and ionized calcium (Ca2+) were determined using calcium-specific electrodes. Saturation with calcium carbonate was calculated by reference to the solubility product of calcite at 37 degrees C. Almost all the samples were found to be supersaturated with calcium carbonate but large variations of the saturation index were observed. In basal samples, obtained during periods of low secretion rate, the mean saturation index (3.35 +/- 3.01) was significantly lower than under secretion (12.10 +/- 5.14) or cerulein (18.01 +/- 8.42). This low basal saturation index, in spite of a high Ca2+ content, was explained by a low bicarbonate concentration (37.6 +/- 18.9 mmol/liter) and a high PCO2 (13.4 +/- 7.5 kPa). In contrast, in juice obtained after hormonal stimulation, PCO2 (4.8 +/ 1.6 kPa) was similar to plasma PCO2 (5.5 +/- 1.2 kPa).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223087 TI - Antibacterial activity of bile salts against common biliary pathogens. Effects of hydrophobicity of the molecule and in the presence of phospholipids. AB - In vitro studies have demonstrated that bile salts have cytotoxic and bacteriostatic properties. The cytotoxic effect of bile salts is reduced when lecithin is added. The effect of lecithin on the bacteriostatic property of bile salts is not known. In this report, we test the hypotheses that (1) the bacteriostatic activity of bile salts is a function of the hydrophobicity of the molecules, and (2) lecithin, by engaging the hydrophobic component of bile salts, attenuates the bacteriostatic property of these molecules. Two common biliary pathogens, Escherichia coli and Enterococcus fecalis, were tested in this experiment. The results demonstrate that hydrophobic bile salts (sodium taurodeoxycholate, sodium deoxycholate) have more significant inhibition on the growth of bacteria when compared with the hydrophilic bile salts (sodium taurocholate, sodium chenodeoxycholate, and sodium tauroursodeoxycholate). When lecithin is added, creating a mixed micellar solution and mimicking the in vivo conditions, the antibacterial activities of even the more potent bacteriostatic bile salts are significantly reduced. The finding that lecithin significantly attenuates the bacteriostatic property of even the hydrophobic bile salts raises questions about the clinical significance of such bacteriostatic effect in vivo; as bile salts in the bile exist in mixed micellar solution. PMID- 8223088 TI - Biliary lithotripsy with a new electromagnetic shock wave source. A 2-year clinical experience. AB - During a two-year study period 170 consecutive patients with gallbladder stones, suitable for lithotripsy, were treated with a new electromagnetic lithotriptor (Modulith) and oral bile acids; 142 patients were treated as outpatients. Sufficient fragmentation were obtained in 94% when 2112 +/- 137 shocks in 211 sessions with an energy setting of 17.8 +/- 0.8 kV were administered. Only 4/170 patients needed transient analgesia. Overall, side effects were transient and mild, but three patients developed biliary pancreatitis, which was treated by endoscopic sphincterotomy in two of them. A total of 67/100 patients were free of stones after one year. Subgroup analysis showed that 80% of the patients (stone diameter 5-20 mm), 64% (20-30 mm) and 65% (multiple stones), respectively, can expected to be free of stones after 12 months. In addition, 25 patients with large, endoscopically not extractable common bile duct stones were treated by lithotripsy with the Modulith. After endoscopic placement of a nasobiliary tube, stone targeting was possible by ultrasonography in 14 patients and by fluoroscopy in another 11 cases. In 23 of the 25 patients (92%) stone clearance by endoscopy was achieved after application of 2516 +/- 565 shocks with an energy preset of 18 kV. One patient refused further endoscopic procedures after successful fragmentation and another required local stone dissolution therapy. Side effects occurred more frequently (P < 0.05) after lithotripsy of bile duct stones than of gallbladder stones, but they were without major clinical relevance. The new lithotriptor Modulith thus enables safe and highly effective lithotripsy of gallbladder calculi on an outpatient basis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223089 TI - Transhepatic topical dissolution of gallbladder stones with MTBE and EDTA. Results, side effects, and correlation with CT imaging. AB - Forty-two patients with symptomatic gallstones (28 women, 14 men, mean age 49.8 +/- 13.2 years) were recruited for contact dissolution therapy. Pretreatment CT scans of the gallbladder were obtained in every patient under standard conditions. For contact dissolution treatment of heterogeneous gallstones or gallstones with attenuation values of more than 50 Hounsfield units, methyl tert butyl ether and bile acid ethylene diaminetetraacetic acid were used in alternating administration at time intervals and durations adapted to the individual tolerance of the patients. In the case of gallstones with mean attenuation values under 50 Hounsfield units, the dissolution therapy was performed with methyl tert-butyl ether alone. In 12 (28.6%) patients a complete dissolution of gallbladder stones could be achieved; 11 patients (26.2%) revealed gallbladder sludge but no radiologically or sonographically visualized residual stone debris. The remaining 19 (45.2%) patients had residual gallstone debris. Shell fragments in three of five rimmed gallstones, seven of eight laminated gallstones, and all densely calcified stones were refractory to contact dissolution therapy. Dissolution rates correlated well with mean attenuation values, whereas no significant correlation was found between stone number and dissolution rates or between stone diameter and dissolution rates respectively. The mean instillation time required for stones with a mean density of more than 50 HU was 17.7 +/- 11.5 hr of bile acid ethylene diaminetetraacetic acid and 5.8 +/- 3.2 hr of methyl tert-butyl ether. In the case of isodense stones, the average instillation time of methyl tert-butyl ether was 12.3 +/- 4.7 hr.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223091 TI - Chronic lymphocytic leukemia with esophageal varices. PMID- 8223090 TI - Alterations of bile acid composition in gallstones, bile, and liver of patients with hepatolithiasis, and their etiological significance. AB - A detailed comparison was made of the bile acid composition in gallstones (brown pigment stones) and paired bile and liver from both affected and unaffected lobes by gallstones, which were taken at operation from 16 patients with hepatolithiasis, with the aim of elucidating whether stone formation is derived from possible local disturbances limited to intrahepatic bile ducts. Brown pigment stones in the intrahepatic bile ducts, most of which were accompanied by bile with high cholesterol saturation, had significantly more cholesterol, and less calcium bilirubinate and bile acid than those found in the extrahepatic bile ducts. Intrahepatic gallstones had significantly lower amounts of secondary and unconjugated bile acids, the bile acids modified by bacterial intervention, than extrahepatic stones. Bile specimens from both affected and unaffected lobes showed significantly increased molar percentages of cholesterol and decreased percentages of bile acids than bile from controls. In contrast, liver specimens from both lobes showed significantly higher concentrations of total bile acids. Secondary bile acids were present in a much lower proportion in bile and liver from both lobes than in bile and liver from controls. On the other hand, unconjugated bile acids were present in a much higher proportion in bile and liver from patients and only in negligible amounts in bile from controls. Furthermore, the plasma levels of mevalonate and those of 7 alpha-hydroxy-4 cholestene-3-one were found to be significantly higher and lower in patients than in controls, respectively, indicating that in hepatolithiasis cholesterol synthesis might increase and bile acid synthesis might decrease in the liver. These findings suggested that alterations of bile acid composition in gallstones, bile, and liver of patients with hepatolithiasis may be attributed to not only secondary changes resulting from local disturbances limited to intrahepatic bile ducts but also possible primary alterations of hepatocyte metabolism, such as bile acid conjugation and primary defects in cholesterol and bile acid synthesis. PMID- 8223092 TI - Hepatotoxicity. AB - Chemical hepatic injury may result from accidental or suicidal exposure to toxic agents in the home or at work or from adverse reactions to medicinal agents. Chemical hepatic injury can lead to acute or chronic syndromes. Acute injury may be cytotoxic, cholestatic, or mixed. Cytotoxic injury is characterized by necrosis, steatosis, or both. Cholestatic injury is characterized by arrested bile flow and may be associated with portal inflammation or may occur in a setting in which there is no evidence of inflammation. Chronic hepatic injury includes chronic active hepatitis, steatosis, phospholipidosis, veno-occlusive disease, several forms of cirrhosis, peliosis hepatis, and hepatic neoplasms. The mechanism for injury may be intrinsic toxicity of the agent, reaction of an unusually susceptible host, or a combination of the two factors. Unusual susceptibility may be the result of immunologic idiosyncrasy (hypersensitivity reaction) or injury from a toxic metabolite (metabolic idiosyncrasy) of the drug. PMID- 8223093 TI - Osteoporosis: pathophysiology, prevention, diagnosis, and treatment. AB - Bone is a living tissue; throughout life, new bone formation coexists with bone resorption. Although a large number of hormones and cytokines modulate osteoblast and osteoclast function, osteoporosis results from any disorder in which bone formation becomes uncoupled from bone resorption. Many disorders are associated with the uncoupling of bone formation and resorption. The most common is loss of gonadal steroid action on bone, as occurs in menopause or in male and female hypogonadism not associated with menopause. Other relatively common causes include primary hyperparathyroidism and endogenous or exogenous hypercortisolism and thyrotoxicosis. A large number of other, less frequent disorders also cause osteoporosis. Treatment of osteoporosis consists first of removing the cause if possible, for example, abolishing hypercortisolism, thyrotoxicosis, or hyperparathyroidism. In menopausal women or hypogonadal men or women, replacement of estrogens or androgens represents effective therapy. Estrogens and androgens given to hypogonadal subjects strikingly reduce bone resorption. For patients with established osteoporosis who either cannot take gonadal steroids or who are not hypogonadal, calcitonin decreases bone resorption and may stabilize bone mass. Estrogen replacement and calcitonin are approved by the Food and Drug Administration for treatment of osteoporosis. Experimental therapies presently include 1,25-dihydroxyvitamin D (calcitriol), bisphosphonates in intermittent treatment regimes, and fluoride in lower dosages than were used in previous studies. The use of fluoride is controversial, and to some extent it has fallen into disrepute. Effective use of any treatment is predicated on understanding the pathophysiology in any particular disease setting. PMID- 8223094 TI - Adult T-cell leukemia with leukemic cell infiltration in the conjunctiva. A case report. AB - A 77-year-old Japanese woman who had suffered from skin eruptions since 1986 was admitted in January, 1990. A diagnosis of adult T-cell leukemia/lymphoma (ATL) was made on the basis of clinical and laboratory data. On admission, erythematous lesions were present in both eyelids. Yellowish-white elevated lesions were found along the limbal conjunctiva, and extended segmentally into the cornea of both eyes. Microscopically, leukemic infiltration into the subepithelial layer of the conjunctiva was observed. Ophthalmic manifestations in ATL have not been well described, because of a little attention paid to them. PMID- 8223095 TI - Negative implant. A retrospective study. AB - Implantation of a negative power intraocular lens is one of the options for surgical correction of high myopia. We studied 36 eyes with a Fechner Worst Claw Lens, implanted in Groningen between March 1987 and November 1991. The preoperative myopia ranged from -7.00 to -30.00 diopters. Twenty one eyes had a follow up period of more than 12 months. The IOL power was calculated with the van der Heyde method. Correction within 1 diopters of emmetropia has been achieved in 55.5% of the cases. Deviation of more than 2 diopters occurred in 25%. In 24 eyes the best corrected postoperative visual acuity improved. Due to the IOL the retinal image increases in size as compared to spectacle correction. In none of the cases complications were encountered during surgery. In the postoperative period one IOL had to be replaced because of an error made in the calculation of the lens power. In one case endothelial decompensation occurred, probably due to a combination of compulsive eye rubbing and a preexisting cornea guttata. The rim of the IOL optic is the part which is the closet to the endothelium. Our recommendation is that one should avoid indentation of the cornea. No other serious complications occurred in this study. PMID- 8223096 TI - The optic nerve in children with craniosynostosis. A pre and post surgical evaluation. AB - Optic nerve (ON) damage especially from papilloedema can be prevented with timely detection. The detailed ocular and radiographic status of 33 unoperated craniosynostosis (CS) was documented prospectively; the ON was also assessed by flash VER and A-scan echography for ON thickness. Fluorescein angiography (FA) was done for all suspected and early papilloedemas. ON involvement diagnosed in 10/33 showed highly significant correlations with multiple suture synostoses (p = 0.004) and exorbitism (p < 0.001). VER latency was increased in all 5 ON atrophies; and also in 1/5 papilloedemas without atrophic changes, urgent surgery in which reversed VER changes, and vision later improved from 6/60 to 6/9. FA was valuable for confirming the clinical diagnosis of the earliest papilloedemas; the FA also returned to normal with immediate therapy along with reversal of the clinical picture. ON echography especially helped in monitoring regression of disc oedema. Ours seems the first report especially in CS where a critical follow up of all above parameters in 21 operated/33 could objectively establish the role of each investigation in conjunction with careful clinical monitoring especially of the ON head. Such comprehensive evaluation should help reduce the blindness often seen in such children. PMID- 8223097 TI - Corneal endothelial damage after Nd:YAG laser anterior capsulotomy. An experimental study on rabbits. AB - The Nd:YAG laser is used primarily in the management of posterior capsular opacification in patients who have had extracapsular cataract extraction. Despite wide clinical use little is known about its damaging effects on the ocular tissues particularly on the corneal endothelium. To evaluate the pathologic changes caused by high powered Nd:YAG laser pulses on the anterior segment of the eye we focused the beam of this laser on the anterior lens capsule in rabbit eyes. Six eyes underwent anterior capsulotomy and three power settings were used (3.4 mJ, 5.8 mJ, 8.6 mJ). The fellow eyes served as controls. We studied acute corneal endothelial changes 6 hours after the capsulotomy using transmission electron microscopy. Results indicate that especially in the high setting the laser energy can cause injury and destruction of endothelial cells and alterations in the Descement membrane. These effects may be crucial of patients with corneas in critical cell number after a cataract surgery. PMID- 8223098 TI - Dynamic register in ocular ultrasonography: combining A- and B-scan techniques. AB - We present the results obtained after studying two groups of patients with dynamic register techniques, using A-scan, standardised A-scan following Ossoinig's procedure and B-scan. One group comprised 50 patients with retinal detachment and proliferative vitreoretinopathy while the other consisted of 15 patients with choroidal melanoma. In the group of patients with retinal detachment and proliferative vitreoretinopathy 90% of cases demonstrated waves with 100% amplitude on A-scan when the ultrasound beam was directed perpendicularly over the detached retina. Likewise in these same 90% of cases we observed a small split in the wave peak during the after movements. Conversely on B-scan 80% of these patients with retinal detachment and proliferative retinopathy showed retinal mobility. In the group of patients with choroidal melanoma rapid spontaneous movements were detected in the internal echoes on A scan and standardised A-scan. Moreover in 3 of the 15 cases (those of greatest size) movement could also be detected on B-scan. PMID- 8223099 TI - Preferential looking: parameters and preliminary data of a new computerized version. AB - A computerized version of preferential looking (CPL) was developed by the authors. Formal parameters (spatial frequency, luminance, contrast of the stimuli; randomized procedure; computerized statistical control) and preliminary binocular acuity results in 69 healthy children (6-36 months) are compared to those of OPL, FPL and ACP version. Low cost, standardized procedure, statistical control of visual acuity estimates and the need of one operator only are among CPL advantages. PMID- 8223100 TI - Preferential looking techniques yield important information in strabismic amblyopia follow-up. AB - Two groups of monocular strabismic patients were followed-up by means of both PL and orthoptic examinations. The first group underwent a full-day occlusion except 1 to 4 hours; the second was occluded less than 4 hours per day. The first treatment regimen proved to be faster in getting pseudo-isoacuity and alternating fixation, but was followed by relative occlusion amblyopia and/or shifting of the dominant eye in three patients. PL techniques proved to be particularly useful in strictly monitoring these patients. PMID- 8223101 TI - Congenital lens coloboma and associated pathologies. AB - Coloboma of the lens is characterized by notching of the equator of the lens. The cause of the condition is faulty development of the zonule. The lens deprived of its normal pull in the defective region is thicker and more spherical as if there were no loss of its substance. We report five cases of lens coloboma and localized zonular defect without any associated systemic anomalies. One case included retinal detachment; another included an iris coloboma; and a third included iris, choroid and optic disk colobomas and retinal detachment with lens coloboma. The remaining two cases were without associated ocular anomalies. The pathogenic mechanisms and relationship of this defect with the differentiation of the tertiary vitreous and development of the zonule is discussed. The defect most likely occurs in the third to fourth months of gestation during differentiation of the tertiary vitreous in response to toxic, inflammatory or genetic factors. PMID- 8223102 TI - Progression of amiodarone induced cataracts. AB - Amiodarone hydrochloride is a potent antiarrhythmic agent recently approved for use by the Food and Drug Administration. Anterior subcapsular lens opacities were observed in seven of fourteen patients treated with moderate to high doses of amiodarone at the Veterans Administration Medical Center in San Francisco in 1982. The present report summarizes the present status of these same fourteen patients ten years later. Anterior subcapsular lens opacities developed or progressed in all patients continuing treatment with this antiarrhythmic agent during the following ten year interval. Although Snellen visual acuities are not decreased, subtle visual impairment is present as measured by contrast sensitivity measurements with and without glare. This decrease in visual acuity is not a contraindication for therapy with this potentially life saving drug. PMID- 8223103 TI - Effects of hemicholinium-3, a photoreceptor and pigment epithelial toxin, on retinal adhesiveness and subretinal fluid absorption. AB - Retinal adhesiveness and subretinal fluid absorption was studied in Dutch rabbit eyes given intravitreal injections of hemicholinium-3 (HC-3) which causes loss of photoreceptor outer segments and retinal pigment epithelial (RPE) damage. After HC-3 administration, some areas of the fundus showed pigmentary changes and others appeared normal. Small, non-rhegmatogenous retinal detachments were made in both areas. Within 2-5 days after HC-3 injection, only in the areas of visible damage, subretinal fluid spread laterally to make very flat retinal detachments, and the fluid absorbed very quickly. At later intervals, absorption was slower than normal, presumably because of scarring and RPE metabolic damage. HC-3 provides an experimental technique for transiently weakening retinal adhesiveness in vivo but its use as a model must account for the effects of both outer segment and RPE damage. PMID- 8223104 TI - Morphological and/or functional imagiology diagnosis of epiphora. AB - Several tests and examinations are necessary for an accurate diagnosis of disorders of the lacrimal drainage system. The AA studied 100 patients with epiphora, divided in two groups. Forty patients, first group (I), aged between 4 and 78 years old (mean age 47.8 +/- 9.6 years), were examined using the classical tests for the study of epiphora, sequential dacryoscintigraphy (SD) and conventional subtraction macrodacryocystrography (CSMD). In the second group (II), 60 patients (aged 45.5 +/- 8.7 years) were studied using the classical tests and by digital subtraction macrodacryocystography (DSMD). The results showed that when functional epiphora is present, there is no difference between a simple clinical approach and sequential dacryoscintigraphy. However, when information is needed regarding choice of surgical approach, DSMD appears to give the best results. PMID- 8223105 TI - Antioxidant activity of plasma from subjects with and without senile cataract. AB - The relationship between overall plasma antioxidant activity and senile cataract was examined in 148 subjects aged 37-90 years. Antioxidant activity of plasma from 52 control and 96 patients with senile cataract was determined by a method which measures the capacity of plasma to inhibit auto-oxidation of bovine brain homogenate. In contrast to previous studies which demonstrated a relationship between decreased levels of antioxidant components in blood and the occurrence of lens opacities, the present study does not show any significant difference between subjects with and without senile cataract. PMID- 8223106 TI - Colored focal visual evoked potentials by cathode ray tube versus scanning laser ophthalmoscope. AB - We compared the focal visual evoked potentials obtained in 52 young subjects with normal vision, evoked by means of three alternating black/color checkerboards generated by a trichromic cathode ray tube (dominant wavelength, 514 nm; colorimetric purity, 0.45) and by means of a scanning laser ophthalmoscope (argon laser beam, 514 nm; colorimetric purity, approximately 1). These three checkerboards, with an area of 3.5 degrees x 3.5 degrees (stimulating the fovea), then with an area of 3.5 degrees x 3.5 degrees with a central exclusion of 1.5 degrees x 1.5 degrees (stimulating the perifoveola) and finally with an area of 1.5 degrees x 1.5 degrees (stimulating the foveola) were presented within a field (8 degrees x 8 degrees) of homogeneous luminance of 170 cd/m2 and 1500 cd/m2, respectively. Their check sizes were 30', with a reversal temporal frequency of 0.75 Hz. The transient focal visual evoked potentials recorded with these three stimuli generated by the two types of stimulators were clearly detected for at least 85% of subjects. Their characteristics (waveform, amplitude and culmination times of the different waves) were comparable, regardless of the stimulator used (cathode ray tube or scanning laser ophthalmoscope). These results suggest that, under these various conditions of luminance and colorimetric purity, the neurophysiologic circuits tested function in identical ways. The focal visual evoked potential signs, now clearly defined by means of stimuli generated by cathode ray tubes, therefore apparently can be applied to the focal visual evoked potential evoked by stimuli generated by the scanning laser ophthalmoscope. PMID- 8223107 TI - Comparison of three methods of estimating the parameters of the Naka-Rushton equation. AB - The Naka-Rushton equation empirically describes the amplitude R of the dark adapted electroretinogram b-wave, as a function of stimulus luminance L, as R/Rmax = Ln/(Ln + Kn). Estimating the three parameters Rmax, n, and K of this function from electroretinogram data is of both experimental and clinical interest. Several different approaches have been developed to accomplish this analysis, but these approaches may derive different estimates of the three parameters. To examine this possibility, we compared the results of three methods of fitting the Naka-Rushton equation to data sets obtained from 30 normal subjects. Two methods were nonlinear curve-fitting programs; the third method involved fitting a regression line to transformed data. The results indicate that solutions provided by these methods have consistent differences, which may be an important consideration when comparing results reported in studies that used different curve-fitting methods. PMID- 8223108 TI - The dark trough in clinical electro-oculography. Influence of preadaptation on amplitudes and latencies. AB - The influence of four different intensities of the preadapting light on amplitude and implicit time of the dark-induced oscillation (dark trough) of the electro oculogram was tested in 10 healthy individuals. The amplitude of the dark trough (normalized to preadapting baseline) was found to be dependent on the intensity of the preadapting light (analysis of variance: p < 0.0001). No correlation could be proven between preadapting light intensity and the implicit time of the dark trough (p = 0.299). The confidence intervals of the implicit time of the dark trough exceeded 14 minutes in all four intensity settings. There was no significant dependence of the absolute baseline values on preadapting intensities (p = 0.236). PMID- 8223109 TI - The second harmonic of the electroretinogram to sinusoidal flicker: spatiotemporal properties and clinical application. AB - The photopic, focal electroretinogram elicited by sinusoidal modulation (8 Hz) of the luminance of a uniform field (flicker electroretinogram) presented in the macular region contains two main components: the first harmonic at the stimulus frequency (1F) and the second harmonic at twice the stimulus frequency (2F). Physiologic and clinical studies have demonstrated that the 1F originates in the outer retina, whereas the 2F has multiple postreceptoral sources, with a contribution of the innermost retina. The 2F, unlike 1F, is selectively altered in amplitude and phase in the early stages of optic neuritis and glaucoma. Both 1F and 2F are altered in maculopathies. An additional property of the focal electroretinogram is that the 2F, unlike 1F, has a very limited summation area (i.e., the function relating 2F amplitude with stimulus area is saturated for central stimulus sizes of the order of the macular region). This overrepresentation of the macular activity suggests that the 2F component of the flicker electroretinogram may provide a specific macular test even for full-field stimuli presented through opacities of the optical media. PMID- 8223110 TI - Visual function in hypermetropia. An electroretinographic and psychophysical study. AB - Dark-adapted retinal function was tested electroretinographically and psychophsysically in patients with severe hypermetropia. These patients were first tested in 1982 and were classified into three electroretinographic categories subnormal, normal and supernormal, according to the amplitudes and the b-wave to a-wave relationships of their dark-adapted electroretinographic responses. These patients were invited for a follow-up examination to examine whether the subnormal electroretinogram represented a stationary or a progressive syndrome, to correlate functional vision to the electroretinographic findings and to determine the changes in refraction and electroretinographic responses that might have occurred during an 8-year period. No significant changes were seen in the amplitudes and b-wave to a-wave relationships of the electroretinographic responses. These observations supported the initial electroretinographic classification of the hypermetropic patients and indicated that the patients belonging to the subnormal group were probably characterized by a stationary defect. The psychophysically determined thresholds at different retinal loci (from 30 degrees nasal to 40 degrees temporal) were within the normal range for all the patients regardless of their electroretinographic characteristics. Thus, the abnormal electroretinographic responses of hypermetropic patients probably did not reflect abnormal retinal function but may be accounted for by changes in the electrical resistances of extraretinal tissues relative to that of the retina itself. PMID- 8223111 TI - Fundus pigmentation and the electroretinographic luminance-response function. AB - Dark-adapted and light-adapted electroretinographic luminance-response functions were recorded from subjects with light or dark fundus pigmentation based on digitized fundus photographs. For dark- and light-adapted electroretinograms, subjects with dark fundi had smaller b-wave amplitudes at all luminance levels. There was no significant difference in b-wave implicit time for the dark-adapted electroretinogram, but there was a significant difference for the light-adapted ERG between the two groups. The results suggest that fundus pigmentation should be considered in the interpretation of electroretinogram results. A possible mechanism for the influence of fundus pigmentation on b-wave amplitude is based on increased resistance associated with melanin. PMID- 8223112 TI - Cone dysfunction and supernormal scotopic electroretinogram with a high-intensity stimulus. A report of three cases. AB - An unusual form of scotopic electroretinogram with a bright white stimulus, which consisted of a rectangular a-wave of normal amplitude and a b-wave of supernormal amplitude, was recorded in three patients with cone dysfunction. In addition to poor visual acuity, abnormal color vision and reduced amplitude of the photopic electroretinogram, these patients showed a 2-log unit elevation of the dark adaptation threshold. Funduscopic examination and fluorescein angiography revealed fine granular pigment disturbances at the macula. The relationship between the response of the dark-adapted electroretinogram versus stimulus intensity was unique to these patients. The b-wave thresholds were elevated by 1 log unit. The b-waves were reduced in amplitude and markedly delayed in implicit time to dim stimuli, but supernormal in amplitude and normal in implicit time to bright stimuli. PMID- 8223113 TI - Intraoperative diagnosis of retinal function. AB - During three-port vitrectomy, we stimulated local areas close to the macula with an intraocular glass fiberoptic. We were able to record a focal electroretinogram in 10 patients with severe diabetic retinopathy, which allowed a more precise evaluation of the retinal function correlated to the pathologic changes caused by the diabetic retinopathy. PMID- 8223114 TI - Visual evoked potentials and electroretinography in brain-dead patients. AB - Visual evoked potentials and electroretinograms were elicited by light-emitting diode stimulation and recorded simultaneously, with cephalic and noncephalic references, in 30 normal subjects and in 30 brain-dead patients. A characteristic pattern was found in the group of patients: when a cephalic reference was used for both visual evoked potentials and the electroretinogram, the a- and b-waves of the electroretinogram were recognized in all patients, and visual evoked responses consisted of waves with inverse polarity, similar morphologic characteristics, the same latency, and less amplitude than those of the electroretinogram. When a noncephalic derivation was chosen for the electroretinogram and visual evoked potentials, electroretinogram did not change in either morphologic features or latency, whereas the visual evoked potential channel showed no response. Only in two cases was it possible to record waves in the visual evoked potential lead with a noncephalic reference, showing a spread of the electroretinogram to the occipital area, with a considerably reduced amplitude. These results suggest that, although contamination of visual evoked potential records by the spread of the electroretinogram to the occipital area could occur, it is easy to confirm the absence of a true cortical visual response in brain-dead patients by means of a noncephalic reference. This pattern clearly confirms that in the visual pathways of brain-dead patients, electrical activity is confined to the retina. PMID- 8223115 TI - Effects of carteolol on the electroretinogram in the perfused cat eye. AB - Carteolol, a nonselective beta-antagonist, was administered intra-arterially in perfused cat eyes. Carteolol increased both photopic and scotopic electroretinogram b-wave amplitude dose-dependently and reversibly, but carteolol failed to induce significant changes in the flow rate of perfusate. This study suggests that carteolol may increase selectively the retinal perfusion flow rate, though it did not reflect the total perfusion flow, or carteolol may have an interaction with retinal beta-adrenergic receptors related to the origin of the b wave. These ideas are supported by carteolol's intrinsic sympathomimetic activity and effects on endothelium of vessels. PMID- 8223116 TI - [Influence of selenium-enriched malt on the elevated cGMP concentrations induced by carcinogen]. AB - Rats were fed with selenium-enriched malt containing 1 ppm selenium. As a parameter, cyclic guanosine-3'5'-monophosphate (cGMP) was measured by RIA in the rats following treatment with AFB1. The results demonstrated that the diet containing high selenium inhibited the elevation of cGMP levels in the liver induced by multiple exposures to AFB1 (250 micrograms/kg, 10 times/2 wks), (P < 0.05). Administration of a large single dose of AFB1 (1 mg/kg i.p.) reduced the liver cGMP contents which might be due to injury to the liver. Selenium-enriched diet could prevent this from happening. In high cancer risk areas: the cGMP level in urine of population with higher selenium concentration were lower than that of population with low selenium concentration. A supplementation of 300 micrograms selenium in the form of selenium-enriched malt cakes daily to the subjects with low selenium concentration in blood for one year, the urinary cGMP excretion was significantly decreased from 0.62 mumol/g to 0.49 mumol/g creatinine (P < 0.05). These findings suggest that diet containing high selenium has a protective effect and may play a potential role in cancer chemoprevention. PMID- 8223117 TI - [Ultra-weak luminescence of serum and urine of bone tumor patients]. AB - The ultra-weak luminescence intensity of serum and urine of 47 cases of bone tumor patients and 17 normal persons are measured by a BPCL-type single-photon ultra-weak luminescence counter. The results showed that the preoperative serum luminescence intensity of the patients was 617.1 +/- 565.5 photon/200 secs and that of urine was 661.8 +/- 283.4, while the serum luminescence intensity of normal group was 236.7 +/- 66 and that of urine was 479.5 +/- 262.8 (P < 0.05). The post-operative serum luminescence intensity of patients was 415.8 +/- 186.6 and that of urine was 311.5 +/- 163.5. The urine luminescence intensity significantly decreased after operation (P < 0.05). The luminescence intensities of serum and urine of bone tumor patients were higher than those of the normal subjects (P < 0.05). The spectrum showed a blueshift with two peaks at 575nm and 665 nm. PMID- 8223118 TI - [Dynamic studies of NK activity, IL-2 production and sIL-2R expression in primary hepatic carcinoma patients before and after operation]. AB - The results of a dynamic study of NK activity, IL-2 production and sIL-2R expression before and after operation in 30 primary hepatic carcinoma patients are reported. The NK activity and IL-2 production of patients were significantly lower, sIL-2R expression was remarkably enhanced than those of controls. There was a period of immune suppression after operation. NK activity and IL-2 production of patients with recurrence were significantly lower and sIL-2R expression was significantly enhanced than than those of patients without recurrence. The period of immune suppression in patients with recurrence was longer than that of patients without recurrence. The results suggest that the changes of NK activity, IL-2 production and sIL-2R expression are closely related to the prognosis of patients with primary hepatic carcinoma. PMID- 8223119 TI - [Long-term results of 60Co HDR after-loading in the treatment of cervical cancer]. AB - From Jan. 1981 to Dec. 1986, 1053 patients with cervical cancer were treated with Cobalt-60 high dose-rate afterloading therapy (Ralstron-20B). All cases have been followed up over 5 years. This paper analysed the 5-year survival rates and complications of cervical carcinoma treated with high dose-rate afterloading therapy. According to the point A dosage, the whole series was divided into four groups: group A 141 patients (65Gy/6 wks), group B 130 patients (60Gy/6 wks), group C 243 patients (55Gy/6 wks) and group D 539 patients (50Gy/5 wks). The dosage of external irradiation in four groups was the same. The 5-year survival rates of the four groups were little better than that of the conventional radium treated group. The incidence of late rectal and bladder reactions in group D was lower than that in group A B C or radium-treated group. These results indicate that the point A dosage of 50Gy/5 wks (dose rate about 100cGy/min) is suitable in HDR intracavitary afterloading therapy of cervical cancer. PMID- 8223120 TI - [Combined use of ondansetron and other anti-emetics to control cisplatin-induced nausea and vomiting]. AB - While ondansetron is effective in the control of nausea and vomiting induced by high dose cisplatin, it has to be given in multiple doses and is very expensive. We designed a regimen of combined use of a single dose of ondansetron and dexamethasone to control acute emesis and combined use of metoclopramide, diphenhydramine and valium to control delayed emesis. The results of this regimen was compared with that of the routine treatment regimen using metoclopramide plus diphenhydramine plus valium. A total of 43 patients were rolled in the cross-over study. Effective control of acute emesis was achieved in 93.6% of the patients with an average of 0.7 emetic episodes. In contrast, effective control of acute emesis was observed in only 18.2% of the patients with an average of 8.1 emetic episodes in those treated with the routine regimen. The regimen increased the emesis control rate by 7.6% and 42.7% on day 1 and 2, respectively as compared to that of repeated administrations of ondansetron alone. The regimen, though not as good as expected, was still better than the routine one for the control of delayed emesis. It deserves recommendation for its better anti-emetic efficacy and lower medical expenses. PMID- 8223121 TI - [Erythropoietin treatment of anemia associated with multiple myeloma (MM) and myelodysplastic syndrome (MDS)]. AB - Anemia is a common complication of patients with multiple myeloma (MM) and myelodysplastic syndrome (MDS). Most of these patients often require blood transfusion. 12 patients, including 7 cases of MM and 5 cases of MDS, were treated with rhEPO 10,000 micrograms three times a week for 15 weeks. The hemoglobin in 6 of 7 cases of MM steadily increased and eventually reached normal level without blood transfusion. The number of erythroid precursors in bone marrow was increased significantly and serum ferritin concentration was decreased gradually during EPO administration. However 5 patients with MDS did not show any response to EPO. The adverse side effects were hardly observed in any patients received EPO treatment. It is suggested that rhEPO is a promising preparation for treating MM-associated anemia rather than MDS-associated anemia. PMID- 8223122 TI - [Treatment of acute promyelocytic leukemia (APL) with all-trans retinoic acid (ATRA): a report of five-year experience]. AB - From January 1986 to April 1991, 100 consecutive patients with APL received oral ATRA at a dose of 60-100 mg/d alone or in combination with chemotherapy. In 84 cases treated with ATRA, 74 (88.1%) achieved CR; in the 16 cases treated with combined therapy, the CR Rate was 75%. Among the 50 patients followed up for a median of 36 months, 10 used ATRA (Group B) as continuation therapy, 10 chemotherapy (Group C), and 30 cases ATRA and chemotherapy alternatively (Group A). The mean survival was 8, 9, 21 months, respectively. For the 29 cases who died, the overall 3-year survival rate was higher in the group A (46.7%) than in the group B and C. ATRA did not provoke or aggravate DIC, nor did it cause bone marrow hypoplasia. The main side effects were dryness of the lip or skin, headache, nausea or vomiting and liver dysfunction. Severe scrotum exfoliative dermatitis with ulceration was seen in one case. In vitro induction of differentiation, GM-CFU, L-CFU assay and cytogenetic studies were performed. The results were discussed together with clinical observation regarding the mechanism of action of ATRA on APL. ATRA used as an inducer of differentiation is an alternative effective drug in the induction of remission in de novo APL as well as in cases in relapse. PMID- 8223123 TI - [Magnetic resonance imaging study of liver metastases]. AB - Eighty-three hepatic metastases from a variety of primary neoplasms were studied with magnetic resonance (MR) imaging. T1 and T2-weighted pulse sequences were employed at a 0.6 T field strength magnet. The results revealed that from multi echo T2 weighted images it seems to be possible to distinguish hypovascular and hypervascular metastatic nodules. Ninety-one percent of target, bull eye and ring signs were present in hypovascular metastases, such as gastrointestinal adenocarcinoma and breast cancer. Eighty eight percent of light bulb sign were present in hypervascular metastases which include endocrine tumors, sarcomas, some lung cancer and renal cell carcinoma. In view of the different vascularity of metastatic nodules, there is less problem to differentiate hypovascular nodules from hemangioma. However it is rather difficult to differentiate the hypervascular metastatic nodules from hemangioma. PMID- 8223124 TI - [Treatment of hepatic neoplasms through extrahepatic collaterals after hepatic artery embolization]. AB - Twenty patients with liver malignant tumors who were treated by hepatic artery occlusion previously, were treated with additional hepatic infusion or embolization though extrahepatic collaterals. Twenty eight courses of hepatic infusion were performed in 14 patients through the right phrenic artery, collaterals of the proper hepatic artery, collaterals of the superior mesenteric and the pancreaticoduodenal arcades, collaterals of the celiac artery, gastroduodenal artery and left gastric artery. Nine hepatic embolization procedures were performed in 5 patients through the right phrenic artery, collaterals of the proper hepatic artery, gastroduodenal artery and collaterals of right renal artery. No complication related to the treatment procedures occurred in this group. The 1-year and 2-year survival rates were 60% and 10%, respectively. PMID- 8223125 TI - [Motor function of the esophagus in patients with esophageal or gastric cancer]. AB - Esophageal carcinoma is one of the most common malignant diseases in China. In order to clarify the pathophysiology of the esophageal motor dysfunction in the disease, a comparative study was done with esophageal manometry and 24 hour pH monitoring in 90 patients with esophageal carcinoma (EC) including 17 cardiac carcinoma (CC) and 56 healthy adult volunteers. The results showed that the resting pressure of the esophagus in patients was lower, particularly during swallowing and Valsalva test, than that of the normal subjects. It indicated that the patients with EC and CC had a hypodynamic esophagus so that a series of abnormal esophageal peristalsis and contractive waves were found. Besides the effect of mechanical obstruction of the tumor itself, it was believed to be a causative factor, at least in part, for the production of the clinical symptoms such as dysphagia and spastic odynophagia occurred during swallowing. Being the lower LESP, the patients with EC and CC presented pathological gastroesophageal reflux which was proved by 24 hour pH monitoring findings. PMID- 8223126 TI - [Eccentric esophagogastrostomy for cancers of gastric cardia and fundus--a report of 122 cases]. AB - Eccentric esophagogastrostomy was performed in 122 patients with cancers of gastric cardia and fundus from 1989 to 1991. Three patients (2.4%) had anastomotic stricture. There were no postoperative anastomotic leakage, gastroesophageal reflux and postoperative death. Since the procedure is performed on an opened stomach under direct vision, the operator can adjust the length of anastomosis. Postoperative X-ray imaging showed that the anastomotic form resembles the anatomical features of gastric cardia and fundus with a larger diameter than that made by routine operation. Therefore, the eccentric esophagogastrostomy may help to reduce the possibility of postoperative anastomotic stricture, prevent gastroesophageal reflux and anastomotic leakage. PMID- 8223127 TI - [Analysis of 15 cases with necrotic perforation and large area necrotic leakage of the gastric wall after operation of esophageal and cardial cancer]. AB - From September 1964 to December 1971 and from January 1986 to December 1991, 15 cases were diagnosed as necrotic perforation and large area necrotic leakage of the gastric wall by iodipin roentgenography, barium meal examination and second operation. It is thought that the pathologic type of anastomotic leakage, the leakage of the anastomotic stoma region, and esophageal and gastric perforation caused by suture cuttings are virtually different from that of necrotic perforation and large area necrotic leakage of the gastric wall, and should be treated differently. The pathogenesis and clinical manifestation of the later two entities are discussed. In our opinion, the maintenance of good circulation of the right gastric and gastroepiploic arteries and veins, careful protection of the stomach wall, and avoiding great tension when the stomach is lifted are of vital importance to prevent the necrosis of gastric wall. In the case of necrotic perforation or large area necrotic leakage of the gastric wall, the early diagnosis is important and early operation is suggested. PMID- 8223128 TI - [Primary hemangiopericytoma of the lung]. AB - Primary pulmonary hemangiopericytoma is a rare form of soft tissue sarcomas. Only six cases were reported in the Chinese literature. Three additional cases treated surgically are now presented, bringing the total to nine and their pathologic and clinical aspects are briefly reviewed. The tumor derives from pericytes and should not be confused with hemangioendothelioma. Most of the patients were asymptomatic and the lesion was detected on routine chest roentgenography. The nine patients had chest pain, fever, cough, dyspnea, hemoptysis or other symptoms, but the symptoms do not differ from those of other lung tumors. Some radiologic features can arouse suspicion of hemangiopericytoma. MRI has a distinct advantage over CT in the tumor imaging. The microscopic morphology is characterized by multiple proliferating capillaries with normal endothelial cells surrounded by proliferating pericytes. For primary pulmonary hemangiopericytoma, the only effective treatment is adequate resection varying from a wedge resection to a pneumonectomy depending on the location and extent of the tumor. PMID- 8223129 TI - [Results of hepatectomy for huge primary liver carcinoma]. AB - From 1964 to 1991, 166 cases of huge primary liver carcinoma (larger than 10 cm in diameter) had been resected in our department. One hundred and forty-four cases were male and 22 cases were female. The tumor was located in the left lobe of the liver in 75 cases, in the right lobe in 75 cases and in both lobes in 16 cases. The tumors were 10 to 14.9 cm in diameter in 109 cases, 15 to 20 cm in 45 cases, larger than 20 cm 12 cases. The largest tumor was 28 x 26 cm in size. In this series, most of the tumors were hepatocellular carcinoma (93.4%). The livers in 125 cases (75.3%) were associated with cirrhosis. Ten cases of them underwent regular hepatectomy. 156 cases underwent irregular hepatectomy. During the operation interrupted matress sutures, hepatic clamp, occlusion of the circulation of the hepatoduodenal ligament and microwave coagulation were employed to reduce blood loss. In this series, 95 cases underwent radical resection and 75 cases palliative resection. The post-operative complications were jaundice and ascites in 18 cases, wounds infection in 7 cases and right pleural effusion in 7 cases. Three cases died within 1 month after hepatectomy with an operative mortality of 1.8%. In our study, 64 cases had been followed up over 5 years. The 5-year survival rate of the radical resection group were 17.5%. All of the 67 patients who underwent palliative resection died 2 to 36 months after hepatectomy with a median survival time of 7 months. The biological characteristics of huge primary liver carcinoma and the approach to improve the prognosis were discussed. PMID- 8223130 TI - [Clinical research on estrogen receptors of breast cancer and endocrine therapy]. PMID- 8223131 TI - [A histopathologic and immunopathologic study of 209 malignant lymphoma cases from four regions of China: Suzhou, Nanjing, Qingdao and Shenyang]. AB - A study was conducted in four regions of China (Suzhou, Nanjing, Qingdao and Shenyang) during the period of 18 months from Dec. 1984 to June 1986, trying to find out the characteristic features of histo- and immuno-pathology of malignant lymphomas in China and relationship with HTLV-1. Fresh specimens were collected from 209 lymphoma patients who were all newly diagnosed. There were 25 cases of Hodgkin's disease (12%), and the other 184 cases of non-Hodgkin's lymphoma (88%). These 184 cases were classified according to the Working Formulation proposed by the National Cancer Institute of USA. It was found that low grade group was 5.4% and intermediate and high grade were 48.9% and 38.0%, respectively. Follicular lymphoma was 4.3% and lymphoma of small lymphocytic type was 2.7%. Fourteen cases (7.6%) were pleomorphic type mimicking adult T-cell lymphoma reported by Japanese. Immunotyping was carried out in 162 cases of non-Hodgkin's lymphoma. The tumors with T-cell markers were 33.9%, B-cell markers 62.4%, and non-T, non-B markers 3.7%. The frequency of T-cell lymphoma cases varied from area to area. Suzhou was 45.8%, Nanjing 57.9%, Qingdao 44.4% and shenyang 11.3%. In the group studied there were only 3 cases showed HTLV-1 positive in serum with pleomorphic morphology and T-cell markers. Our findings indicate that composition of histopathologic types of malignant lymphoma in China is different from that in western countries, and the incidence of T-cell lymphoma is high in certain areas of the lower reaches of the Yangzi River.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223132 TI - [Amplification of int-2 gene in primary esophageal carcinoma and fetal esophageal carcinoma induced by N-methyl-N-benzylnitrosamine (NMBzA)]. AB - Amplification of int-2 gene in primary esophageal carcinomas and adjacent tissues, fetal esophageal carcinoma induced by NMBzA and in fetal esophageal epithelium treated with NMBzA was detected. Amplification of int-2 gene was observed in 23 of 52 esophageal carcinoma (44.2%), 7 of 18 adjacent tissues (38.9%). We also found that the amplification of int-2 in fetal esophageal carcinoma induced by NMBzA and in fetal esophageal epithelium treated with NMBzA for 1 month. These results demonstrate that NMBzA activates possibly the int-2 in human esophageal epithelium to induced the esophageal carcinoma, indicating that the NMBzA may play an important role in the development of esophageal cancer. PMID- 8223133 TI - [The expression of C-myc, N-ras mRNA and its relations to the differentiation of preneoplastic altered hepatocytes]. AB - The expression of C-myc and N-ras mRNA was studied in preneoplastic hepatic lesions induced by multiple small doses of diethylnitrosamine (DEN). Dot blot analysis showed that the expression of C-myc and N-ras mRNA was in elevated levels during the early stage of hepatocarcinogenesis. In situ hybridization in tissue sections demonstrated that the over-expression of C-myc and N-ras mRNA was clearly seen in some of the preneoplastic altered liver cell lesions. In comparison of these over-expressed oncogene lesions with the expression of carbamyl phosphate synthetase I (CPS I), known as a differentiation marker of hepatocytes, in serial sections revealed that almost all of the foci with over expressed C-myc and/or N-ras mRNA also showed low expression of CPS I mRNA, which means a low differentiation nature of the liver cells. These findings suggested that the overexpression of C-myc and N-ras are closely correlated to the differentiation state of altered liver cells. PMID- 8223134 TI - Induction of morphological and enzymic differentiation in rat pheochromocytoma PC12h cells by stable erbstatin analogues. AB - Erbstatin, a tyrosine kinase inhibitor, is known to induce transient differentiation in rat pheochromocytoma PC12h cells. The authors tested seven newly synthesized stable erbstatin analogues for their ability to induce neurite formation and acetylcholine esterase in PC12h cells. Among the inhibitors tested hexyl 2,3-dihydroxycinnamate most efficiently induced neurite formation in PC12h cells. Although this analogue is much more stable than erbstatin, the induction by it was again transient. In addition, the analogue weakly induced acetylcholine esterase activity in PC12h cells. Thus hexyl 2,3-dihydroxycinnamate is a potent inducer of morphological differentiation in PC12h cells. PMID- 8223135 TI - The role of serotonin brain receptors in the analgesic effect of phenazone. AB - The effects of treatment with para-chloro-phenyl-alanine (PCPA) (100 mg/kg i.p. for 4 days) were studied on the hot-plate test and on brain 5-HT binding in phenazone treated rats. Phenazone per se induces analgesia in the hot-plate test and decreases the number of cortical and pontine 5-HT binding sites A pre treatment with PCPA prevents both the analgesic effect and the reduction of 5-HT binding sites caused by phenazone. These data suggest that the brain serotonin system may play a role in phenazone-induced antinociception. PMID- 8223136 TI - Effect of oral activated charcoal on vancomycin clearance in rabbits with acute renal failure. AB - The effect of oral administration of activated charcoal on total body clearance of vancomycin administered intravenously (7.5 mg/kg) has been studied in normal rabbits and rabbits with induced renal failure. Gastric intubation of a single dose (10 g) of activated charcoal to normal rabbits did not produce a statistically significant effect on any pharmacokinetic parameter for vancomycin. The mean vancomycin clearances were (mean +/- s.d.) 80.82 +/- 6.8 and 75.24 +/- 9.61 ml/h/kg with and without activated charcoal administration, respectively. To examine whether renal failure would influence the effect of activated charcoal and enhance the systemic clearance of vancomycin, uranyl nitrate was used (0.75 mg/kg, i.v.) to induce acute renal failure in rabbits. The derived pharmacokinetic parameters of vancomycin were consistent with renal failure. No significant differences were observed in any of the calculated pharmacokinetic parameters between the control and charcoal treated rabbits. The lack of effect may be attributed to the large molecular weight of vancomycin. PMID- 8223137 TI - Pharmacokinetics of lamotrigine in patients with renal impairment: influence of haemodialysis. AB - The pharmacokinetics of a single 100 mg oral dose of lamotrigine, a new anticonvulsant drug, were studied in six healthy volunteers and in twenty patients with various degrees of renal impairment. Six of these patients were regularly haemodialysed. Lamotrigine concentrations in plasma and urine were measured by the HPLC method. The mean peak plasma concentrations of lamotrigine, tmax, volume of distribution and total clearance were not significantly modified by the degree of renal impairment. The elimination half-life of lamotrigine was approximately 25 h in subjects with normal renal function and 50 h in uraemic patients. These are very large variations in uraemic patients and the results were not statistically significant. Renal clearance of lamotrigine is significantly reduced. Urinary elimination of unchanged and conjugated lamotrigine was reduced in uraemic patients. Thus it seems necessary to treat carefully patients with a very severe renal insufficiency since very large variations in pharmacokinetics were found. A 100 mg oral dose every two days is recommended if creatinine clearance is below 10 ml/min. Haemodialysis shortened the elimination half-life from 59.6 +/- 28.1 h during the interdialysis period to 12.2 +/- 6.4 h during the dialysis period; 17% of the dose was extracted by haemodialysis. PMID- 8223138 TI - Enantioselective effects of levodropropizine and dropropizine on psychomotor functions in normal volunteers: a placebo-controlled, double-blind comparative study. AB - Levodropropizine is the l-isomer of dropropizine, a racemic drug widely used as a cough suppressant. Compared with the racemate, levodropropizine retains equal antitussive activity but exhibits considerably lower central nervous system (CNS) depressant effects in animal models. In order to assess whether the same differential pharmacodynamic profile also applies to man, a double-blind placebo controlled study was carried out to investigate the effects of single oral doses (60 and 120 mg) of levodropropizine and dropropizine on subjective alertness (scored on visual analogue scales), general tolerability and psychomotor function tests (cancellation, tapping, choice reaction times and critical flicker fusion frequency) in ten normal volunteers. Treatments were administered in random sequence at intervals of at least one week, evaluation procedures being carried out at times 0, 1, 2, 3, 4, 6 and 8 h after dosing. Following intake of a 60 mg levodropizine dose, subjective effects and objective estimates of psychomotor function were superimposable to those recorded after placebo. There was a trend for 60 mg dropropizine and 120 mg levodropropizine to produce detrimental effects at occasional evaluations, although the changes associated with these treatments could not be differentiated from placebo on the basis of most subjective scores and psychomotor function tests. Conversely, administration of 120 mg dropropizine was consistently associated with subjective CNS impairment and with reduced performance (compared to baseline) in recognition time, critical flicker fusion thresholds and possibly tapping rate, for up to three hours after dosing. These data are consistent with evidence that racemic dropropizine adversely affects central nervous system function to a greater extent compared with the levo isomer. PMID- 8223139 TI - Treatment of gonococcal infections with a single 250 mg intramuscular injection of trospectomycin sulphate vs ceftriaxone sodium. AB - Trospectomycin sulphate is a new, more potent analog of spectinomycin, which is active in vitro against penicillin-sensitive and penicillin-resistant strains of Neisseria gonorrhoeae. This study was designed to determine the bacteriologic and clinical efficacy as well as safety of a single intramuscularly administered 250 mg dose of trospectomycin sulphate in the treatment of uncomplicated gonorrhoea (cervical, urethral, pharyngeal and anal). Ceftriaxone sodium was used as a comparator antibiotic in a single 250 mg intramuscular dose. Seventy-four patients (36 women and 38 men) were evaluable in the trospectomycin treated group and 40 patients (22 women and 18 men) in the ceftriaxone treated group. The overall bacteriologic cure rate was 98.6% (73/74) for trospectomycin and 95% (38/40) for ceftriaxone. Bacteriologic failures were observed among women 1/36 (2.8%) treated with trospectomycin and 2/22 (9.1%) treated with ceftriaxone. The overall clinical success rate (clinically cured plus clinically improved) was 90.5% for trospectomycin and 100% for ceftriaxone. Adverse events were reported rarely in both groups. Less than 10% of patients complained of pain and/or tenderness at the injection site for both drugs; one patient developed a generalized, pruritic rash which occurred three days after administration of trospectomycin and resolved within six days. In conclusion, a single dose of 250 mg i.m. trospectomycin appears to be at least as effective and safe as a single dose of ceftriaxone in the treatment of uncomplicated gonorrhoea. PMID- 8223140 TI - Cefixime in the treatment of enteric fever in children. AB - Cefixime in a dose 20 mg/kg/day, orally, divided into two doses 12 h apart for a minimum of 12 days, was administered to 50 children with proven S. typhi septicaemia. Forty four of the patients were infected with strains of S. typhi resistant to multiple antibiotics including chloramphenicol, ampicillin and trimethoprim-sulfamethoxazole. All patients responded rapidly to treatment and were cured clinically and bacteriologically. Fever subsided within a mean of 5.3 days (range 3-8 days). Only two of the 50 patients treated relapsed during the 8 week follow-up period. No serious adverse reactions attributable to the drug were observed. Cefixime proved to be an effective oral drug in this open treatment trial and was associated with minimal side effects. It may provide a therapeutic alternative to the treatment of Salmonella infection with organisms multi resistant to the standard drug regimens. Its oral formulation may provide an efficient alternative to parenteral therapy in less severely ill patients who can tolerate oral feeding. PMID- 8223141 TI - Protective role of propionyl carnitine in vascular disorders experimentally induced by endothelin (ET-1) serotonin and K-carrageenin. AB - Propionyl carnitine, a derivative of carnitine, has metabolic and cardiovascular effects similar to carnitine but with more pronounced peripheral haemodynamic activity. In these experiments we propose to prove that the administration of propionyl carnitine could prevent the experimental tail thrombosis in the rat induced by endothelin (ET-1), serotonin and K-carrageenin. In this new test of experimental thrombosis, propionyl carnitine was able to reduce the extent of tail thrombosis in a more significant manner than that of carnitine. A possible explanation of this antithrombotic effect of propionyl carnitine is its capacity to counteract the vasoconstrictor activity of endothelin modulating the release of prostanoids induced by endothelin itself. PMID- 8223142 TI - Cefdinir: in vitro activity study and effect of human serum. AB - The in-vitro activity of cefdinir (Cl-983, FK482), an orally absorbed aminothiazole cephalosporin, was compared with that of penicillin, ampicillin, amoxycillin, amoxycillin/clavulanic acid (2/1), cefaclor, cefuroxime, cefixime, cefotaxime, vancomycin and erythromycin against 370 clinical isolates of Gram negative and Gram-positive bacteria. Cefdinir was highly active against Staphylococcus aureus and S. epidermidis, inhibiting 90% of the strains at doses of 0.25 and 0.5 mg/l respectively. However, cefdinir was not active against methicillin-resistant S. aureus (range 16- > 128 mg/l). The respiratory pathogens Moraxella catarrhalis, Streptococcus pneumoniae, and S. pyogenes were also susceptible (MIC90 < or = 0.5 mg/l), but against Enterococcus spp. cefdinir displayed no useful activity. The common members of the family Enterobacteriaceae were susceptible (MIC90 < or = 1 mg/l), but those possessing chromosomal beta lactamases were more resistant (MIC90 2-8 mg/l). The presence of human serum had little effect on MICs of cefdinir. These results indicate that cefdinir exhibited a wide spectrum for an oral cephalosporin and support its possible clinical use against susceptible pathogens in infections of the skin, soft tissue, respiratory and urinary tracts. PMID- 8223143 TI - Microbicidal effects of ozone solution on methicillin-resistant Staphylococcus aureus. AB - Methicillin-resistant Staphylococcus aureus has become an important problem in chemotherapy in the past decade because of its resistance to commonly used disinfectants. The usefulness of ozone solutions to disinfect against strains of this bacterium has been evaluated. Ozone strongly oxidizes the cell walls and cytoplasmic membranes of bacteria. The microbicidal effect of ozone solutions took place within the first five seconds of treatment. An ozone concentration 1.5 times that needed to kill the one methicillin-sensitive strain tested was sufficient to kill the resistant clinical isolates. Such an ozone solution could be used to disinfect medical instruments and similar equipment. Ozone solutions should be useful in reducing the number of bacterial infections caused by inadequate disinfection against new resistant strains. PMID- 8223144 TI - Protective action of L-carnitine and coenzyme Q10 against hepatic triglyceride infiltration induced by hyperbaric oxygen and ethanol. AB - This investigation was conducted to ascertain whether administration of L carnitine and coenzyme Q10 could protect from the experimentally-induced hepatic lipid infiltration and glutathione content decrease in rats exposed to hyperbaric oxygen and prolonged alcohol administration. The results indicate that administration of L-carnitine and coenzyme Q10 in association reduces damage induced by chronic alcohol poisoning and hyperbaric oxygen. This protective action is more marked when L-carnitine and coenzyme Q10 are administered together. The combined complementary biochemical activity of these two compounds is discussed. PMID- 8223145 TI - Protective effect of Swertia chirata against indomethacin and other ulcerogenic agent-induced gastric ulcers. AB - The effect of Swertia chirata has been studied on experimentally induced gastric ulcers in rats. The ethanolic extract of chirata significantly reduced the intensity of gastric mucosal damage induced by indomethacin and necrotizing agents. It produced a significant decrease in gastric secretion in pylorus ligated rats. The extract inhibited acetylcholine-induced contraction of guinea pig ileum, suggesting its anti-cholinergic activity. Pretreatment of rats with the extract significantly prevented ethanol-induced gastric wall mucus depletion and restored the non-protein sulfhydryl (NP-SH) content in the glandular stomachs. These findings support the use of chirata for the treatment of gastric ulcers in traditional medicine. PMID- 8223146 TI - Thrombosis induced by endothelin (ET-1) and carrageenin in rats treated with indomethacin and propionyl carnitine. AB - Propionyl carnitine can prevent endothelin and carrageenin-induced thrombosis in the rat's tail. This protective action of propionyl carnitine is probably related to its capacity to increase the synthesis of PGI2 prostaglandins and, subsequently, the amount of prostaglandins released by endothelin. Indomethacin, a well known prostanoid synthesis blocker, is unable to inhibit the protective activity of propionyl carnitine on endothelin-induced thrombosis after prolonged propionyl carnitine administration, but only a part of this protective activity after propionyl carnitine acute administration. Subsequently, other factors in addition to prostanoids seem to be involved in protective mechanisms of propionyl carnitine on endothelin-induced thrombosis. PMID- 8223147 TI - [Ectopic expression and the significance of HLA-class II antigens in the myocardium in patients with dilated cardiomyopathy]. AB - Expression of HLA-class II antigens (DQ, DP) in the myocardium of patients with different heart diseases and normal controls was studied with indirect immunofluorescence (IIF). The positive rates in different groups were as follows: dilated cardiomyopathy (DCM, 12/13,+- ), rheumatic heart disease (2/4, +-+ + +), congenital heart diseases (1/14,+), left atrial myxoma (0/1) and normal controls (1/8,+). The immunofluorescent area was in the endocardium and endothelial cells of the blood vessels (VIII factor related antigen positive areas in dual staining of the myocardium). There were significant differences between the positive rates of DCM and those of other heart diseases including rheumatic heart disease, congenital heart diseases and left atrial myxoma (chi 2 cor = 15.3, P < 0.001), as well as between DCM and normal controls (chi 2 cor = 9.9, P < 0.01). The expression of HLA-class II antigens is highly selective. Ectopic expression has been postulated to be responsible for autoimmune diseases. So it can be used as a specific marker for autoimmunity in DCM. PMID- 8223148 TI - [A new method for the localization of atrial septal puncture by using atrial septal electrocardiogram]. AB - A new method for the localization of atrial septal puncture was reported, which is the key technique for percutaneous balloon mitral valvuloplasty. When the exploratory electrode was laid against the fossa ovalis floor (FOF), the atrial septal electrocardiogram registered only a slight or no injury curve; when the exploratory electrode pressed on any other area of the atrial septum or atrial walls, a bizarre monophasic injury curve was elicited in the atrial septal electrocardiogram. Based on different patterns of the atrial septal electrocardiogram between the muscular atrial septum and FOF, the sites of atrial septal puncture were successfully localized by using a Brockenbrough needle as an exploratory electrode in all of the 10 cases without any complication. By using this technique, no cardioangiography is needed to show the outline of the left atrium and also the bilateral projection catheterization bed. So this method is convenient, practical and precise. PMID- 8223149 TI - [Effect of intravenous atropine on the induction of paroxysmal supraventricular tachycardia]. AB - Sixty nine patients with the history of paroxysmal supraventricular tachycardia (PSVT) or possible history of PSVT failed to have PSVT provoked by transesophageal atrial pacing (TEAP). After intravenous administration of 0.02 mg/kg atropine, 42 patients had PSVT provoked during the second TEAP. The electrophysiologic parameters showed that atropine could increase the inducibility of PSVT. The main effect was to decrease the anterograde AV node effective refractory period (ERP) or fast pathway ERP and provide an available condition for the reentry of impulses. The conclusion is that atropine could definitely increase the diagnostic value of TEAP. PMID- 8223150 TI - [Clinical study on the relationship between erythrocyte ATPase activity and lipid peroxidation in essential hypertension]. AB - The changes of erythrocyte ATPase and superoxide dismutase (SOD) activity, intraerythrocytic ionic concentration and plasma lipid peroxide (LPO) were observed in 59 patients with essential hypertension (EH). The results showed that the Na(+)-K(+)-ATPase, Ca(2+)-Mg(2+)-ATPase activity and SOD were significantly lower, Ca2+ and Na+ concentration and plasma LPO were significantly higher in EH than in normals. A significant linear relationship was found between the decline of ATPase activity and changes of LPO and SOD. Both LPO and Ca(2+)-concentration were positively correlated with mean arterial blood pressure. These findings suggest that lipid peroxidation, through inhibiting ATPase activity and increasing intracellular Ca2+ concentration, plays an important role in the pathogenesis of EH. PMID- 8223151 TI - [Platelet calcium influx and changes of phosphatidyl inositol]. PMID- 8223152 TI - [Incision of the accessory pathway of left heart free wall under direct vision in patients with Wolff-Parkinson-White syndrome. Report of 4 cases]. PMID- 8223153 TI - [Diagnosis of II degree atrioventricular block by ECG during atrial fibrillation]. PMID- 8223154 TI - [Clinical analysis of 76 cases of persistent ventricular tachycardia]. PMID- 8223155 TI - [An analysis of dynamic electrocardiogram in patients with sinus bradycardia and sick sinus syndrome]. PMID- 8223156 TI - [Effects of captopril on myocardial ultrastructure in diabetic rats]. AB - Diabetic rat models were established with intraperitoneal injection of streptozotocin. The animals were randomly divided into four groups: Group 1, normal control (n = 5); Group 2, diabetic control (n = 4); Group 3, diabetics treated with PZI (n = 5); Group 4, diabetics treated with PZI and captopril (n = 5). Specimens taken from free wall of the left ventricle were observed under electron microscope after 10-week treatment, and the volume density (Vv), numerical density on area (Na), numerical density (Nv) and mean volume (V) of mitochondria were measured with stereological quantitation. The results showed that the myocardial changes in Group 4 were much less severe than those in Group 3 and similar to those in Group 1. There was significant difference in Nv (P < 0.05) but not in Vv, Na and V between Group 4 and Group 1. While the changes of Group 3 were relatively obvious. As comparing Group 3 with Group 1, there were significant differences in all of the four parameters (P < 0.05-0.01). It is implied that the renin-angiotensin system may be involved in the pathogenesis of diabetic cardiomyopathy, since captopril can improve and reverse the cardiomyopathy of diabetic rats. PMID- 8223157 TI - [Effects of calcitonin-gene related peptide on local rheology of normal and ischemic myocardium in dogs]. PMID- 8223158 TI - [A study on the dynamic effect of nifedipine on red blood cell cytoplasmic free calcium in hypertensive patients]. PMID- 8223159 TI - [The effect of indapamide on phosphate metabolism in hypertensive patients]. AB - Twenty-four patients were studied to assess the efficacy and principle of indapamide in hypertensive patients previously treated and unsatisfactorily controlled with either angiotensin-converting enzyme (ACE) inhibitor or atenolol therapy. The result showed a satisfactory anti-hypertensive effect of indapamide. Plasma phosphate concentrations were markedly reduced and no significant variance in plasma K+, Na+ and Ca+ concentration after two weeks of indapamide therapy (2.5 mg, one day). Thus indapamideinduced reduction of blood pressure could be mediated by an effect of this drug on phosphate balance. PMID- 8223160 TI - [Hypotensive effect of neuropeptide Y in substantia nigra]. AB - The central hypotensive effect of neuropeptide Y (NPY) has been investigated by microinjection of the peptide into substantia nigra (SN) of anesthetized normotensive rats. NPY microinjection into SN produced dose-dependent falls in arterial blood pressure but no change in heart rate statistically. Microinjection of cholecystokinin octapeptide or 0.9% saline into SN did not elicit prominent change in arterial blood pressure and heart rate. The results suggest that NPY in SN exerts a distinctive hypotensive effect in certain degree. PMID- 8223161 TI - [An immunohistochemistry study on the histologic origin of Aschoff body in 27 cases of rheumatism]. PMID- 8223162 TI - [The advances in the techniques of interventional therapy of occlusive arterial diseases]. PMID- 8223163 TI - [Using myocardial perfusion tomography to estimate infarct size and to determine prognostic value in patients with acute myocardial infarction]. AB - Single-photon emission computerized tomography with Tc-99m-MIBI was used to measure the infarct size in 41 patients with acute myocardial infarction (AMI), among them 11 patients (accounting for 27%) were dead or recurred with perfusion defect > 31% in 6 cases including 3 deaths and 2 recurrences. The perfusion defect was between 11%-30% in 12 patients, including one death and two recurrences. The perfusion defect was 10% in 23 patients, including 3 with recurrence. During the period from 6 months to 2 years following-up, the total death and patients with recurrence were 11/41 (27%) cases. The results showed that the prognosis of patients with AMI was closely related to the size of myocardial perfusion defect, and those with perfusion defect > 31% had a poorer prognosis. PMID- 8223164 TI - [Application of central-line-chord method to study left ventricular wall motion in left ventriculogram]. AB - The left ventricular wall motion in left ventriculogram of 71 patients with coronary artery disease was analysed in our laboratory with central-line-chord method. 55 of 71 patients had ventricular wall motion abnormalities (77.46%). The regional wall motion abnormality (RWMA) often occurred in the areas of ventricles with poor blood supply from the stenosed coronary artery (CCA). When both LCA and RCA had stenosis, abnormal area would show in chords 10 to 28 or chords 40 to 80, when LAD stenosed, chords 10 to 30 or chords 52 to 720, and when CFX stenosed, then chords 64 to 80 were involved. The results also suggest that RWMA would appear when EF was still within normal limits. Thus, RWMA provides earlier prediction of abnormal left ventricular function than LVEF. PMID- 8223165 TI - [Treatment of acute myocardial infarction with oral captopril. A randomized, double blind and placebo controlled pilot study]. AB - A randomized, double blind and placebo-controlled pilot study was done on the use of converting enzyme inhibitor captopril in 98 patients in the early stage of acute myocardial infarction (AMI) with the purpose of testing the feasibility and justifiability of a large clinical trial on such therapy. The results of the study were as follows. (1) The randomization in this study was quite satisfactory. The interruption of the treatment protocol for more than 24 hours and the patients drop out rate were only 5.1% each, and no side effects of captopril were observed. These indicated that such study could be carried out on large scale. (2) Captopril slightly lowered blood pressure (average 10.2/4.5mm Hg after 2 hours of oral 6.25mg) with minimal change of heart rate, but it did not increase the incidence of sustained hypotension. (3) Captopril decreased the angiotensin II level significantly after 2 hours of ingesting 12.5mg. (4) Early captopril treatment tended to reduce the incidence of cardiac free wall rupture, the severity of congestive heart failure and cardiogenic shock, therefore tended to reduce the early mortality of AMI (3/43 captopril group VS 8/55 placebo group). PMID- 8223166 TI - [A multicenter cooperative study on prevention and treatment of unstable angina associated with acute myocardial infarction and cardiac death]. PMID- 8223167 TI - [Calcitonin gene-related peptide and thrombolysis]. AB - After thrombolytic therapy with urokinase (UK) plasmaimmunoreactive calcitonin gene-related peptide (CGRP-ir) in patients with acute myocardial infarction (AMI) was increased rapidly, and 2 hours after treatment with UK the plasma CGRP-ir reached peak value which was 5 times higher than that before thrombolytic therapy. Fourteen hours after therapy with UK the plasma CGRP-ir gradually decreased to the initial level before treatment. In rat model with abdominal aorta thrombosis induced by FeCl3-injured vascular endothelium, it was found that change in plasma CGRP-ir of the rats after treatment with UK was similar to that of AMI patients, and that administration of CGRP alone had no recanalization effect on the thrombotic blood vessel. However, treatment with CGRP synergically potentiated the UK-induced vascular recanalization in a dose-dependent manner. And antagonist of CGRP (CGRP8-37) significantly attenuated the recanalization action of UK. The results suggest that release of CGRP by tissues could play an endogenous protective role in thrombolytic therapy with UK, and exogenous administration of CGRP might be useful clinically as an assistant of vascular recanalization. PMID- 8223168 TI - [A study on the hemodynamic effect of cedilanid in the treatment of acute myocardial infarction]. AB - For long time, there has been controversy on the use of digitalis in acute myocardial infarction (AMI). Hemodynamic studies before and after the use of cedilanid, simple QRS scoring, changes on the score of AMI and cardiac arrhythmia were analysed on 24 cases of AMI. Results of the pre- and post-medication values were improved as following: (1) Cardiac index (CI) were 2.34 +/- 0.2 and 3.21 +/- 0.26L/min/m2 two hours after medication (P < 0.05). (2) Left ventricular stroke work index (LVSWI) were 31.2 +/- 3.4 and 40.1 +/- 2.2 g-m/beat/m2 two hours after medication (P < 0.01). (3) Pulmonary capillary wedge pressure (PCWP) were 19.5 +/ 2.4 and 13.2 +/- 2.1 mmHg two hours after medication (P < 0.01). (4) Myocardial oxygen consumption triple index (TI) were 182000 +/- 14000 and 142000 +/- 12000 two hours after medication (P < 0.01). (5) There was no change or with only trivial aberration on systemic vascular resistance (SVR) and perfusion pressure of the coronary arteries (CCP). (6) Simple QRS scoring on electrocardiogram resulted that the size of infarcted areas were 4.02 +/- 0.3 points before and 3.01 +/- 0.23 points after the medication (P < 0.01), there was neither ventricular premature beats (VPB) nor increase of any other serious cardiac arrhythmias in the electrocardiogram care and record in 72 hours. From the hemodynamic studies, the beneficial effect of cedilanid is greater than its adverse effect. It is concluded that digitalis can be safely and effectively used in the treatment of AMI. PMID- 8223169 TI - [Mechanism of the alternation cycle length phenomenon during intravenous verapamil for termination of atrioventricular reciprocating tachycardia]. AB - The electrophysiologic-pharmacologic test of verapamil on 33 patients with the Wolff-Parkinson-White syndrome with atrioventricular reciprocating tachycardia (AVRT) was studied. During intravenous verapamil for termination of AVRT, the alternating cycle length phenomenon was observed in 13 cases (39%). On the basis of the observations we attributed the phenomenon to alternating conduction between early and late phase of the AV nodal relative refractory period due to the AV nodal refractory period lengthened by verapamil except a few patients in whom verapamil unmasked the presence of concealed accessory pathways or dual AV nodal pathways as hypothesized by some scholars. PMID- 8223170 TI - [Spectrotemporal mapping of signal-averaged electrocardiogram for identification of patients with sustained ventricular tachycardia]. AB - Frequency analysis of multiple segments (spectrotemporal mapping, STM) of signal averaged ECG (SAECG) considerably enhanced the detection of late potentials in patients with VTs/VF, but has not yet been verified in large samples. In this study, the modified protocol of STM, using the time when vector summed QRS forces last decreased to 40 microV at 25-250Hz filtering (40 microV/25-250Hz) as the onsets of the window, was performed in 119 normal subjects (group I), 26 patients with VTs/VF (group II), and 202 patients with organic heart diseases of different causes not accompanied by VTs/VF. The minimal normal factors (NF) of individual X, Y, Z leads were compared, significant differences were found between each 2 groups. 95% confidential values of minimal NF from group I were calculated, and NF < 33% on any lead was recommended as abnormal, ie, LP present. With this standard, 81% of the patients with VTs/VF, 94% of the normal subjects and 80% of the patients without VTs/VF were correctly discriminated despite the presence of intraventricular block (IVB, including bundle branch block and nonspecific intraventricular block). In conclusion, the method of STM of SAECG using terminal 40 microV/25-250Hz as window onsets offered promise for identification of patients with VTs/VF, and had better clinical applicability than time domain analysis for patients with IVB may not be excluded during STM. PMID- 8223171 TI - [Idiopathic restrictive cardiomyopathy--clinical and histopathologic manifestations in two cases]. PMID- 8223172 TI - [The natural history of sick sinus syndrome]. AB - Ninety one cases of sick sinus syndrome (SSS) have been traced clinically for an average term of 5 years and 4 months. The result shows that the natural history of SSS is relatively long, no one of these cases died of arrhythmia during the investigation, and pacemaker is the first choice for patients with apparent severe symptoms. PMID- 8223173 TI - [Application of transesophageal color Doppler flow imaging in decision making during mitral valve operation]. PMID- 8223174 TI - [Spontaneous echo contrast in atrium with rheumatic heart disease by transesophageal echocardiography and its potential relation to thrombus]. AB - The prevalence of atrial spontaneous echo contrast (SEC) and mural thrombus were evaluated in 154 consecutive patients undergoing transesophageal echocardiography (TEE) and transthoracic echocardiography (TTE). By TEE, SEC was observed in 106 patients (68.8%). Of them, 103 were seen in left atrium and only 3 were seen in right atrium. No SEC and mural thrombus were seen by TTE. Patients with mitral stenosis were in 82 of 154 patients, SEC was seen in 74 of those 82 patients (90%). Mechanical stenosis in mitral orifice is a predominant factor. Our analysis showed significant relation between the presence of SEC and atrial fibrillation (P < 0.005); left atrial enlargement (P < 0.001) and cardiac days function (P < 0.005). Of the 106 patients with SEC, 41 (38.7%) were in sinus rhythm and only 8 with the left atrial diameter > 60 mm. Left atrial mural thrombus was noted in 52/154 (33.6%), 42 of those 52 were patients with SEC (80%), which also suggest a significant relation between the presence of SEC and mural thrombus (P < 0.025). Thus SEC is a common phenomenon in patients with rheumatic heart disease. Its relation to thrombus formation and prognostic significance requires further evaluation. PMID- 8223175 TI - Psychological treatment for anxiety--an alternative to drugs? PMID- 8223177 TI - Patient choice in managing cancer. PMID- 8223176 TI - Acitretin, a new retinoid. PMID- 8223178 TI - Vitamin K supplements in the newborn. PMID- 8223179 TI - [Nursing models for the future]. PMID- 8223180 TI - [Dealing with nursing models in some European countries]. PMID- 8223181 TI - [Who is succeeding whom?--or: In-hospital nursing succeeds outpatient nursing. A current commentary on the theme: Nursing Services Insurance]. PMID- 8223182 TI - [Is further education as community nurse necessary?]. PMID- 8223183 TI - [Execution and introduction of a patient discharge record]. PMID- 8223184 TI - [Home care of the aged and family medicine]. PMID- 8223185 TI - [Observations and experiences of a clerical commissioner for work with the aged]. PMID- 8223186 TI - [A day care center for mentally retarded seniors]. PMID- 8223187 TI - [Realization of the care legislation in a social work department]. PMID- 8223188 TI - [He who will not help himself can not be helped at all. The importance of self help groups for the individual and for society]. PMID- 8223190 TI - [Parent self-help group cooperating with a pediatric department]. PMID- 8223189 TI - [Work place: self-help group]. PMID- 8223191 TI - [Questions of liability in outpatient nursing]. PMID- 8223192 TI - [What is a nursing problem?]. PMID- 8223193 TI - [Education towards nursing predicaments? A plea for supervision during education]. PMID- 8223194 TI - [Basics in nursing research]. PMID- 8223195 TI - [The indications for transesophageal echocardiography after cerebral ischemia]. AB - Possible intracardiac sources of emboli were looked for by transoesophageal echocardiography (TOE) in 271 patients (149 men, 122 women; mean age 60 +/- 11 years) a mean of 12 +/- 8 days after suffering an episode of cerebral ischaemia. Left atrial thrombi were revealed in 9 patients: they were more frequent in those with atrial fibrillation (odds ratio [OD] 25.2; P < 0.0001) or known cardiac disease (OD 3.5; P = 0.06). Using the two factors together, the 9 left atrial thrombi could be predicted in 96 patients, while in the remaining 175 patients without cardiac abnormalities no left atrial thrombi were found. Overall, TOE is not an essential investigation additional to transthoracic echocardiography to exclude left atrial thrombi in patients without heart disease who are in sinus rhythm. But in those with atrial fibrillation and/or organic heart disease, TOE can in many instances facilitate the indication for anticoagulation. PMID- 8223196 TI - [Necrotizing hepatitis with a fatal outcome after carbimazole therapy]. AB - A 74-year-old woman developed weakness, lack of appetite and abdominal swelling 7 months after starting treatment with carbimazole (10 mg/d for 10 weeks) and, subsequently, radioiodine for hyperthyroidism. Physical examination revealed generalized oedema and ascites. Computed tomography showed a liver of normal size but infiltrated by nodules up to 4 cm in diameter. Erythrocyte sedimentation rate was raised and there were abnormal concentrations of haemoglobin, total proteins, liver enzymes and creatinine, as well as decreased platelet and white cell counts. Thyroid function was normal. Viral and autoimmune diseases were largely excluded. Liver biopsy showed a severe, highly active hepatitis with parenchymal necroses, large-drop fatty infiltration and intralobular granuloma-like inflammatory reactions, as well as lympho-histiocytic inflammation of the portal areas. The most likely cause was the carbimazole treatment. In addition to symptomatic treatment she received prednisone (1 mg/kg), because an allergic diathesis could not be excluded. But she died of hepatic failure 6 weeks after admission. PMID- 8223197 TI - [Is chronic dermato- or polymyositis misdiagnosed as "weakness of old age"?]. AB - Two women, aged 72 (case 1) and 77 (case 2) were referred for neurological diagnosis because of progressive muscular weakness, for 4 and 18 months, respectively, which had finally led to dysphagia and required mechanical ventilation. The cause of the disease in case 1 was classical dermatomyositis. Creatinine kinase concentration, never previously measured, was 950 U/l. In case 2 there was polymyositis, previously not considered as she had a diabetic polyneuropathy and muscle enzyme concentration was normal. In both cases the correct diagnosis was speedily confirmed by electromyography and muscle biopsy. Immunosuppressive treatment was largely successful in reversing the symptoms (case 1: initially 500 mg methylprednisolone daily, reduced to 50 mg daily within 6 weeks; case 2: 500 mg methylprednisolone daily for 1 week, then 100 mg daily plus 150 mg azathioprine with maintenance dosage of 20 mg glucocorticoid daily).- These two cases demonstrate that, particularly in the elderly, dermatomyositis and polymyositis should be considered in the differential diagnosis of progressive general weakness. PMID- 8223198 TI - [Magnetic resonance spectroscopy of the heart. Its clinical importance]. PMID- 8223199 TI - [Nitrates]. PMID- 8223200 TI - [Airway resistance measurement]. PMID- 8223201 TI - [Pericardial effusion and puncture]. PMID- 8223202 TI - [Lysis therapy in phlebothrombosis]. PMID- 8223203 TI - [Manometric studies of the esophagus in healthy subjects of different age groups]. AB - Oesophageal manometry was performed under standardized conditions in 100 healthy persons of different ages (54 males, 46 females; mean age 43.1 [18-84] years). They were grouped by age in decades (< or = 29 years: n = 23; 30-39 years: n = 18; 40-49 years: n = 19; 50-59 years; n = 20; and > or = 60 years: n = 20). The mean resting pressures at the lower oesophageal sphincter ranged among the different age groups from 18-22 mm Hg (total group 19 +/- 7 mm Hg), the mean contraction force at the distal tubular oesophagus ranged from 77-100 mm Hg (84 +/- 35 mm Hg), in the proximal tubular oesophagus from 53-58 mm Hg (56 +/- 22 mm Hg), and the resting pressures at the upper oesophageal sphincter from 34-45 mm Hg (37 +/- 17 mm Hg). There was no statistically significant difference between the various age groups for any of the motility parameters. It is concluded that age does not influence the findings in comparative studies of healthy persons and patients of differing ages. PMID- 8223204 TI - [Histoplasmosis of the skin as an initial opportunistic infection in AIDS]. AB - A 55-year-old homosexual Indonesian (last stay in Indonesia 2 years previously), known to be HIV positive since 1986, developed desquamating, in part ulcerating, skin eruption over the face and shoulder region. On admission his temperature was 38.2 degrees C, erythrocyte sedimentation rate 72/95 mm, white cell count 3.100/microliters, and the CD4 cell count 30/microliters. Examination of lung, oesophagus, stomach, duodenum and colon for possible opportunistic infections was negative. Fundoscopy revealed an infiltrate in the right eye with destruction of the vitreous. Skin biopsy suggested histoplasmosis, confirmed by culturing H. capsulatum varietas capsulatum. It is likely that this was the reactivation of a latent, previously symptom-free infection, in this case the first opportunistic infection in the presence of AIDS. For 30 days he received infusions of amphotericin B (initially 0.1 mg/kg daily, after the 5th day 0.5 mg/kg), resulting in rapid healing of the skin lesions. Subsequently he has received (for 6 months so far) itraconazole, 400 mg daily, without further complications. PMID- 8223205 TI - [Recurrent venous thromboses in hypoplasia of the vena cava inferior and factor XII deficiency]. AB - The cause of recurrent pelvic and leg venous thromboses in a 24-year-old man was found to be a combination of two rare anomalies, hypoplasia of the hepatic, prerenal segment of the inferior vena cava and factor XII deficiency (factor XII activity 38%, its antigen 39% of normal), the latter considered a risk factor for thromboembolism. Subsequent fibrinolysis was not successful. No thromboembolic phenomena occurred during the following 16 months of oral anticoagulation with phenprocoumon. When this treatment was discontinued at the patient's behest, there was a recurrence on the contralateral side. Anticoagulation was resumed and has continued now for 1 1/2 years without recurrence. The patient has been largely free of symptoms. Permanent anticoagulation thus seems unavoidable in this case. PMID- 8223206 TI - [The primary immune response]. PMID- 8223207 TI - [The clinical picture and genetics of tuberous sclerosis]. PMID- 8223208 TI - [The coincidence of latex and food allergies]. PMID- 8223210 TI - [High blood pressure in young adults]. PMID- 8223209 TI - [The HLA-DRB1 gene and the severity of rheumatoid arthritis]. PMID- 8223211 TI - [Long-term electrocardiography in the idiopathic QT syndrome]. AB - The value of long-term electrocardiographic (ECG) monitoring was assessed in 14 patients (8 males, 6 females; mean age 21 [17-30] years) with the idiopathic long QT syndrome (LQTS), 14 healthy subjects of the same age serving as controls. Twelve patients had the typical history of syncopes or sudden cardiac death among family members; seven patients had a history of syncope, while four patients had been successfully resuscitated. None had associated cardiac disease. Among the group with LQTS the rate-corrected QT interval at rest was 498 + 56 ms, in the control group 412 +/- 30 ms (P < 0.005). Resting and maximal heart rates on exercise were similar in the two groups. The rate-corrected QT interval on exercise was significantly longer in the LQTS patients (P < 0.001). In the control group the maximal heart rate in the long-term ECG was significantly higher (144 +/- 28/min) than in the LQTS patients (128 +/- 17/min; P < 0.01). The long-term ECG recorded abnormal findings in five patients: torsade-de-pointes tachycardia in two, T-wave alternans in two, and bradycardia resulting from intermittent sinoatrial block in one. No abnormal findings were recorded in the control group (P < 0.03). PMID- 8223212 TI - [The temperature dependence of fibrinolysis with streptokinase]. AB - In an in-vitro study standardized test thrombi were incubated at different temperatures between 25 degrees and 43 degrees C in a constant test mixture in which all reagents necessary for fibrinolysis including streptokinase were contained in excess. At each temperature step the dilution of the standard streptokinase solution (1:1000 IU/ml) was measured at which the test thrombus dissolved in exactly 15 min (clot lysis test). The highest fibrinolysis activity (dilution of 1:1140 of the standard) was obtained at 40 degrees C, lower activities having been recorded at both higher and lower temperatures (43 degrees C:1:1000; 37 degrees C:1:900; 34 degrees C:1:550; 30 degrees C:1:255; 25 degrees C:1:90). Related to the optimal temperature of 40 degrees C (100%), the fibrinolytic activity at 25 degrees C was only 7.9%. In view of the axial temperature gradients existing in the limbs, the internal temperature in their peripheral parts lies well below the optimal temperature for fibrinolysis. This situation should be taken into account during fibrinolysis procedures for thrombotic occlusions in the limbs. PMID- 8223213 TI - [Acute cytomegalovirus gastritis after kidney transplantation. Its diagnosis by the polymerase chain reaction, antigenemia assay and immunohistochemistry]. AB - To diagnose possible cytomegalovirus (CMV) infection in a 64-year-old man after renal transplantation, polymerase chain reaction (PCR), pp65 antigenaemia assay (pAA) and virus isolation in cell culture were routinely performed on a weekly basis. The PCR obtained virus DNA in peripheral blood lymphocytes for the first time in the fifth week. Two weeks later the patient complained of feeling unwell with abdominal pain and vomiting on eating. Two days later he developed a fever up to 38 degrees C and nocturnal sweats. Gastroscopy revealed marked antral gastritis which histologically showed typical cytomegalic "owl-eye" cells. The pAA was clearly positive and the cell culture started in the fifth week now showed a cytopathogenic effect. CMV gastritic having been diagnosed treatment consisted of 175 mg ganciclovir intravenously twice daily for 10 days. He became symptom-free after two days. The only side effects were thrombocytopenia down to 67,000/microliters and a rise in transaminase activities, changes which regressed later. To ensure early diagnosis and treatment of any CMV infection, specific virus diagnostic tests should be routinely undertaken after transplantation of organs from CMV-positive persons into CMV-negative patients. PMID- 8223214 TI - [The clinical picture and diagnosis of bronchiectasis]. PMID- 8223215 TI - [The therapy of bronchiectasis]. PMID- 8223216 TI - [Coronary angioplasty in unstable angina pectoris: immediately or after an interval?]. PMID- 8223217 TI - [Tetanus diseases, inoculations and adverse inoculation effects in the Federal Republic of Germany 1971 to 1990]. PMID- 8223218 TI - [Medical society dues for the academic medical profession. The judgement of the Federal Administrative Court of 26 January 19913]. PMID- 8223219 TI - [Lyme borreliosis]. PMID- 8223220 TI - [The therapy of primary gastric lymphomas]. PMID- 8223221 TI - [Dietary calcium and kidney stone formation]. PMID- 8223222 TI - [Color-coded duplex sonography in the diagnosis of renal venous thromboses in the early phase following kidney transplantation]. AB - A total of 91 colour-coded duplex Doppler ultrasound scans were performed in the early phase following renal transplantation in 53 out of 109 patients (48.6%) between 1. 1. 1992 and 15. 3. 1993. The indications for the investigation were primary failure of functional improvement in the transplanted kidney or secondary worsening of function. In five of the 53 patients (ten scans) constant retrograde flow in the segmentary arteries provided specific evidence of renal vein thrombosis, this diagnosis being confirmed histologically following nephrectomy. The Pourcelot index was raised (RI > 0.72) in all five cases. The RI was also raised in 35 of 53 patients; however, continuous retrograde diastolic flow was absent. The results of renal biopsy and the clinical course both excluded renal vein thrombosis in these cases. The colour Doppler ultrasound scans were unremarkable in 13 patients. The comparison group consisted of 151 patients in the later posttransplantation phase (> 2 months), in whom 171 colour Doppler ultrasound scans showed no signals associated with renal vein thrombosis. In agreement with the literature, these results show that colour-coded duplex Doppler ultrasound should be regarded as the definitive diagnostic procedure for renal vein thrombosis after transplantation, and if positive findings are obtained further invasive diagnostic procedures can be avoided. PMID- 8223223 TI - [Pneumonia due to a rare atypical Mycobacterium in AIDS]. AB - A 38-year-old man, HIV-positive for 6 years, developed fever and cough with deterioration in his general state. Chest radiography demonstrated an infiltration in the left upper lobe and computed tomography showed a septated cavity. Three bronchioalveolar lavages over 4 weeks recovered Klebsiella, Candida, Pseudomonas and Staphylococcus in the lavage fluid. Acid-fast rods were not found in any of the microscopic preparations. His clinical condition and the radiological findings deteriorated despite appropriate antibiotic administration. A further cavity occurred in the right upper lobe and the inflammatory infiltrations extended further. Although no acid-fast organism had been demonstrated, tuberculostatic treatment was begun (daily 300 mg isoniazid, 600 mg rifampicin, 900 mg streptomycin, 2 g pyrazinamide). His general condition and the radiological findings rapidly improved. Four weeks after culturing the lavage fluid atypical Mycobacterium xenopi was isolated. This case illustrates the difficulty of diagnosing an atypical mycobacterial infection. It takes time and effort, but it is of great importance because up to 50% of patients with AIDS contract such infection. Early and appropriate treatment will significantly improve quality of life and life expectancy. PMID- 8223224 TI - [The clinical course and therapy of massive theophylline poisoning]. AB - With suicidal intent a 72-year-old man swallowed 5.8 g aminophylline in a non retard solution. The theophylline plasma level on admission was 120 mg/l. He had to be intubated when respiratory arrest occurred. Within the first hour he developed cerebral seizures, polymorphous ventricular premature systoles, atrial fibrillation with an irregular ventricular rate and, finally, recurrent episodes of ventricular fibrillation with prolonged circulatory shock (heart rate 120 140/min with a systolic blood pressure of 60 mm Hg for 3 hours) and severe metabolic acidosis (potassium 2.28 mmol/l, phosphate 0.21 mmol/l, pH 7.03, base excess -20.8 mmol/l). He was treated with massive fluid replacement (6.2 l in the first 12 hours), electrolyte substitution to counteract the marked hypokalaemia and hypophosphataemia, repeated defibrillation and antiarrhythmic drugs (lidocaine 240 mg/h and metoprolol twice 5 mg), as well as anticonvulsive treatment (diazepam, 10 mg twice, followed by midazolam 5 mg/h). Detoxication measures consisted initially of gastric lavage followed by high-dosage enteric administration of charcoal (210 g over 36 h), as well as haemoperfusion for 4 h. Full recovery was achieved and the patient was discharged in good health after 3 weeks. PMID- 8223225 TI - [The indications for laparoscopy in internal medicine]. PMID- 8223226 TI - [Aspects of pharmacotherapy in old age]. PMID- 8223227 TI - [Stress studies in pneumology]. PMID- 8223228 TI - [Influenza infection and antibiotic therapy]. PMID- 8223229 TI - [Computed tomographic diagnosis in stroke]. PMID- 8223230 TI - [Heparin-induced extracorporeal LDL precipitation and ACE inhibitors]. PMID- 8223231 TI - [A comparison of the gastroduodenal tolerance of ticlopidine and acetylsalicylic acid]. PMID- 8223232 TI - [Calcium therapy in women following menopause]. PMID- 8223233 TI - [Reflections on 100 years of animal breeding as mirrored in the Deutsche Tierarztliche Wochenschrift]. AB - In 1893 a major component of veterinary practice derived from 4 million horses and from farm animals, whereas thousands of dogs were even slaughtered every year, especially in Saxony and Silesia. Nowadays companion animals figure as an element of ever growing importance for the practitioner. Teaching and research adapted themselves to this altered situation, research funding and professional politics not yet so completely. In former days veterinarians even presided the licensing committees for breeder animals--today the animal breeding act merely mentions them in connection with institutions of AI and ET. Misdevelopments during the Nazi period are discussed as possible causes. In analogy to refugee problems after World War II, in this decade crowding effects again challenge the profession, resulting from increasing numbers of students and European migrations. PMID- 8223234 TI - [Literature review of the use of albumin solutions in human medicine for the purpose of investigating the possibility of an indication for the therapy of dehydration conditions in veterinary practice]. AB - The use of Albumin solutions as a homologous agent is sufficient in diseases corresponding to hypovolemia because of its long half-time, water binding capacity and rare side-effects. Young animals have a high ratio of extracellular/intracellular space and are severely affected by the loss of fluid. They have to be treated with sufficient amounts of plasma expanders over a long period to fill up the vascular system. This treatment maintains the microcirculation and stimulates the kidney activity thereby preventing shock. The application of electrolyte solutions as an only substitution of existing deficits ist needed after the general health situation of the animal has improved and the animal is drinking water. Albumin has several advantages compared with crystalloid and other colloid solutions and can therefore be used successfully in veterinary practice. However, it is necessary to find better ways to produce high quality Albumin solution for a reasonable price. PMID- 8223235 TI - [The content of iron, copper and zinc in 12 tissues of female swine in 11 age groups from birth to a body weight of 200 to 265 kg]. AB - In female pigs in the age of one day (body mass: 1.89 +/- 0.20 kg) to that of on the average 1123 days (b. m: 200 to 265 kg) in 11 age-groups the weight of 12 different tissues as well as their content in Fe, Cu and Zn were analysed. The age dependent changes of the content are described and discussed. PMID- 8223236 TI - [The effect of a Bordetella live vaccine on the occurrence and manifestation of atrophic rhinitis suum and the aerogenous infection burden in field strains of the agent]. AB - A field study was carried out in a large scale unit for swine breeding and fattening with the object to influence the high morbidity rate of Atrophic Rhinitis and pneumonia with the help of a Bordetella live vaccine. The results show that it is possible to decrease the infectious pressure by B. bronchiseptica and to reduce the pathomorphological signs of Atrophic Rhinitis in consequence of the application of the live vaccine. The pathologic-anatomical investigations of nasal turbinates in immunized slaughtered fattening pigs show a significant lower morbidity concerning Atrophic Rhinitis and a higher percentage of pigs without changes at conchae nasales and septum nasi. We find also a lower contamination of the air with B. bronchiseptica field strains during vaccine application. The results also explain that a high infectious pressure by B. bronchiseptica and the possibility of communication between unvaccinated and vaccinated groups of pigs counteract a better efficiency of the vaccine. The decrease of the morbidity rate of Atrophic Rhinitis appears so much more important because toxicogenic Pasteurella multocida strains were isolated from nasal swabs of vaccinated pigs during the investigations. But these strains influenced the Atrophic Rhinitis frequency only accidentally. All results as a whole point out that in pig houses with a high animal density one has to pay more attention to virulent B. bronchiseptica strains than it was been done till now. PMID- 8223237 TI - [Divided animal protection: study of a natural science and human science evaluation of the federal German animal welfare law]. AB - The meaning of the word "animal" in section 1 of the animal protection law of the Federal Republic of Germany encompasses every animal of the zoological classification (from protozoa to primate). This is stressed by the word "fellow creature" for an animal, because vertebrates and non-vertebrates all are part of creation. Further in section 1 the life as well as the well-being of the individual animal are governed. The ethical and philosophical ways of thinking which led to the protection of the individual life of every animal of the zoological classification are not compatible with the generally accepted use of the word "animal" (protozoa to primate), because individual life protection cannot be practiced for the majority of animal species--the non-vertebrates. However, to concentrate on the protection of the life of the individual animal of only a few species in our daily life is emotionally acceptable but arbitrary. Animal protection has to be focused on the protection of well-being of animals, which are able to suffer, predominantly vertebrates. PMID- 8223238 TI - Detection by cultural isolation of bovine virus diarrhoea (BVD) virus following field infections in cattle and their fetuses in Turkey. AB - Cultural isolation, identification and biotype characterization of BVD virus is based on monolayer tissue culture techniques using mainly organs of, e.g., bovine fetuses. These might have experienced a transplacental transmission and are therefore of no use for tissue cultures produced, e.g., from kidneys of such fetuses. In order to rule out BVD virus infections, fetuses must be tested for BVD virus, viral antigen or specific nucleotide sequences. BVD virus and other ruminant pestiviruses have been detected in Turkey by cultural isolation. In bovine fetuses BVD virus has not been demonstrated so far. Cultural isolation was attempted in the present investigation in 50 randomly selected pregnant cows which were brought to an abattoir. Of these cows serum samples were assayed for neutralizing antibodies to BVD virus and 40 (80%) found positive with titres > or = 1:40. In contrast fetuses 120 days of ontogenic age and therefore supposedly immunocompetent were derived from seropositive dams (n = 16) but did not show any neutralizing antibodies to BVD virus. Leucocyte fractions of peripheral blood samples of the 50 cows as well as 353 fetal blood and organ samples were inoculated and submitted to three passages in fetal calf kidney cell cultures before testing for BVD viral antigen by immunofluorescence technique. BVD virus was thus detected in the fetus of one seropositive cow and also in the leucocyte fraction of one seronegative dam including organs of her fetus. The number of fetuses found positive for BVD virus amongst fetuses collected in abattoirs corresponds well to figures gained by other investigators outside Turkey.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223239 TI - [Enrofloxacin (Baytril)--an alternative for psittacosis prevention and therapy in imported psittacines]. AB - It was the aim of the presented work to test Enrofloxacin under field conditions of compulsory prophylaxis and treatment of psittacosis, as JUNG (1993) had already demonstrated the effectiveness of this antibiotic against Chlamydia psittaci. During official quarantine 22 groups comprising 2536 birds were treated with Baytril. With suitable feed like steamed maize and a dose of 500 mg/kg Enrofloxacin the mean blood values ranged from 0.9 to 4.1 micrograms/ml within the different species. Application via drinking water can only be advised as an alternative in sick birds with reduced food intake. Duration of excretion (limit was the minimal inhibitory dose of Enrofloxacin with Chlamydia psittaci, 0.125 micrograms/ml was depending on the uptake of Enrofloxacin and correlated with serum levels. With mean serum levels of 1.0 micrograms/ml the minimal inhibitory dose for Chlamydia psittaci was attained in all birds after 14 days. The tissue concentrations measured in Patagonian conures surpassed serum levels several fold, with the exception of brain tissue. Out of 22 quarantine groups only two were naturally infected with Chlamydia. One group of 196 Senegal parrots could only be cured from chlamydial infection after substituting their normal mixed feed with medicated maize containing 1000 mg/kg Enrofloxacin. In 50 per cent of the Salmonella infected groups Salmonella could again be grown from faecal samples after the end of treatment. For the treatment of psittacosis mean blood levels of 0.5 micrograms/ml are imperative and duration of treatment should not be below 14 days. Satisfactory hygienic conditions provided Baytril is a valuable therapeutic drug against psittacosis. PMID- 8223240 TI - [Mesothelioma in a cow]. PMID- 8223241 TI - [Toxoplasmosis epizootic in a fattening swine herd]. AB - In a fattening pig herd comprising 80 animals 13 were affected with clinical manifest toxoplasmosis presumably due to cat feces in the food. Seven pigs died, one was killed in extremis. The clinical signs were anorexia, apathy, fever, cyanosis, dyspnoea and partly hind limb weakness. One pig that died 3 days after onset of clinical signs had widespread necroses in liver and lymphatic organs. Single tachyzoites were detected by immunoperoxidase technique. One pig dying on day 11 and another euthanatized the same day showed severe desquamative and interstitial pneumonia and marked non suppurative meningoencephalomyelitis as well as necrotic foci mostly infiltrated with mononuclear cells in liver, adrenals and lymphatic organs. In all affected organs toxoplasmas were demonstrated histologically and immunohistologically. Every 2 surviving pigs were killed 40 days and 20 weeks, respectively, after recovery. These animals had high antibody titers against toxoplasma gondii and showed severe non suppurative meningoencephalitis and toxoplasma cysts in the brain without preceding clinical signs. In the lymphatic organs a marked hyperplasia was observed. PMID- 8223242 TI - [Effects of dehydrocholic acid (Biliton) on metabolism and liver function in cows]. AB - Dehydrocholic acid (Biliton) was given to 9 cows with a predisposition for the fat mobilization syndrome in daily doses of 5.5 g each. This was done two weeks after parturition and the results were compared with those from 9 untreated cows. Five other cows suffering from ketosis or indigestion symptoms were treated too. Decreased concentrations of liver lipids, free fatty acids (FFA), bilirubin, beta OH-butyrate and urea as well as increased glucose in blood plasma indicated a favourable action of Dehydrocholic acid on metabolism and liver function. We did not observe a significant influence on milk and reproduction parameters. The use of Dehydrocholic acid is recommended for use in liver disturbances. PMID- 8223243 TI - Mice homozygous for a targeted disruption of the proto-oncogene int-2 have developmental defects in the tail and inner ear. AB - We derived mice that carry a targeted insertion of a neor gene in the int-2 (Fgf 3) proto-oncogene coding sequences. The mutation was found to be recessive and mice that were homozygous for the insertion did not often survive to adulthood. The mutant mice had defects in the development of the tail and inner ear that could be correlated with disruption of int-2 expression in the posterior primitive streak and hindbrain or otic vesicle. While the tail phenotype was 100% penetrant, we found that the inner ear phenotype had reduced penetrance and variable expressivity. The variable expressivity could not be attributed to variability in the genetic background of the mutant allele or to leaky expression from the mutant allele. Thus, we conclude that even in a uniform genetic background, stochastic variation in the expression of a developmental circuit can result in dramatic differences in phenotypic consequences. PMID- 8223244 TI - Homeostatic balance between dorsal and cactus proteins in the Drosophila embryo. AB - The maternal-effect gene dorsal encodes the ventral morphogen that is essential for elaboration of ventral and ventrolateral fates in the Drosophila embryo. Dorsal belongs to the rel family of transcription factors and controls asymmetric expression of zygotic genes along the dorsoventral axis. The dorsal protein is cytoplasmic in early embryos, possibly because of a direct interaction with cactus. In response to a ventral signal, dorsal protein becomes partitioned into nuclei of cleavage-stage syncytial blastoderms such that the ventral nuclei have the maximum amount of dorsal protein, and the lateral and dorsal nuclei have progressively less protein. Here we show that transgenic flies containing the dorsal cDNA, which is driven by the constitutively active hsp83 promoter, exhibits rescue of the dorsal- phenotype. Transformed lines were used to increase the level of dorsal protein. Females with dorsal levels roughly twice that of wild-type produced normal embryos, while a higher level of dorsal protein resulted in phenotypes similar to those observed for loss-of-function cactus mutations. By manipulating the cactus gene dose, we found that in contrast to a dorsal/cactus ratio of 2.5 which resulted in fully penetrant weak ventralization, a cactus/dorsal ratio of 3.0 was acceptable by the system. By manipulating dorsal levels in different cactus and dorsal group mutant backgrounds, we found that the relative amounts of ventral signal to that of the dorsal-cactus complex is important for the elaboration of the normal dorsoventral pattern. We propose that in a wild-type embryo, the activities of dorsal and cactus are not independently regulated; excess cactus activity is deployed only if a higher level of dorsal protein is available. Based on these results we discuss how the ventral signal interacts with the dorsal-cactus complex, thus forming a gradient of nuclear dorsal protein. PMID- 8223245 TI - The Drosophila gene pointed encodes two ETS-like proteins which are involved in the development of the midline glial cells. AB - Glial-neuronal cell interactions at the ventral midline are necessary for the proper elaboration of commissures in the embryonic CNS of Drosophila. In particular, migrating midline glial cells are required for the separation of segmental commissures. During this process the glial cells recognize specific neuronal cells at the midline, they migrate posteriorly along their cell processes and thereby separate the segmental commissures. The gene pointed (pnt) is required for this glial-neuronal cell interaction, as loss of function mutations lead to a change in the migration behavior of the midline glial cells. As a consequence, anterior and posterior commissures do not become separated and appear fused. Molecular analysis of pointed has revealed two differently spliced types of transcripts, which are encoded in a region extending over 55 kb of genomic sequence. In the CNS both transcript classes are expressed in cells of the midline, including the midline glial cells. Sequence analysis of cDNA clones corresponding to both transcript types reveals two different pointed proteins which share an ETS domain common to a number of transcription factors related to the vertebrate ets oncogene. Furthermore, one pointed protein form contains an additional domain of homology of approx. 80 amino acids in length, which is shared by only a subset of the ETS protein family. PMID- 8223246 TI - Developmental origin of segmental identity in the leech mesoderm. AB - Segmentation in the leech embryo is established by a stereotyped cell lineage. Each of the 32 segments arises from homologous, bilaterally symmetrical complements of mesodermal and ectodermal blast cell clones. Although segments are homologous, they are regionally differentiated along the longitudinal body axis. Various segments display idiosyncratic ensembles of features, which constitute discrete segmental identities. The differentiation of segment-specific features, such as the mesoderm-derived nephridia, genital primordia and identified Small Cardioactive Peptide immunoreactive neurons, reflects a diversification of the developmental fates of homologous blast cell clones. We have investigated whether segment-specific differentiation of homologous mesodermal blast cell clones depends on cell-intrinsic mechanisms (based on the cells' lineage history) or on cell-extrinsic mechanisms (based on the cells' interactions with their environment) in embryos of Theromyzon rude. For this purpose, we first mapped the segment-specific fates of individual mesodermal blast cell clones, and then induced mesodermal clones to take part in the formation of segments for which they are not normally destined. Two types of ectopic segmental position were produced: one in which a mesodermal blast cell clone was out of register with all other consegmental cells and one in which a mesodermal blast cell clone was out of register with its overlying ectoderm, but was in normal register with the mesoderm and ectoderm on the other side of the embryo. Mesodermal blast cell clones that developed in either type of ectopic segmental position gave rise to segment-specific features characteristic of their original segmental fates rather than their ectopic positions. Thus, the development of segmental identity in the leech mesoderm is attributable to a cell-intrinsic mechanism and, either before or soon after their birth, mesodermal blast cells are autonomously committed to segment-specific fates. PMID- 8223247 TI - Induction of floor plate differentiation by contact-dependent, homeogenetic signals. AB - The floor plate is located at the ventral midline of the neural tube and has been implicated in neural cell patterning and axon guidance. To address the cellular mechanisms involved in floor plate differentiation, we have used an assay that monitors the expression of floor-plate-specific antigens in neural plate explants cultured in the presence of inducing tissues. Contact-mediated signals from both the notochord and the floor plate act directly on neural plate cells to induce floor plate differentiation. Floor plate induction is initiated medially by a signal from the notochord, but appears to be propagated to more lateral cells by homeogenetic signals that derive from medial floor plate cells. The response of neural plate cells to inductive signals declines with embryonic age, suggesting that the mediolateral extent of the floor plate is limited by a loss of competence of neural cells. The rostral boundary of the floor plate at the midbrain-forebrain junction appears to result from the lack of inducing activity in prechordal mesoderm and the inability of rostral neural plate cells to respond to inductive signals. PMID- 8223248 TI - Genetic analysis of the Drosophila cdc2 homolog. AB - We have identified mutations in the Drosophila cdc2 gene. The recessive lethality of these mutant alleles was rescued after P-element-mediated transformation with a genomic cdc2 fragment. Sequence analysis of amorphic alleles revealed non conservative exchanges in evolutionary conserved positions. These alleles caused lethality at the larval-pupal interphase due to the absence of imaginal tissues. Embryonic lethality resulted when the maternal Dm cdc2 contribution was reduced through the use of a temperature-sensitive allele. Dm cdc2 function, therefore, is essential for cell proliferation throughout development. Dm cdc2 function is clearly required for mitosis, but no evidence for a requirement in S-phase was obtained. The reversible block of the mitotic proliferation which was observed in the PNS of mutant embryos occurred exclusively in the G2-phase. Moreover, while the mitotic proliferation of imaginal cells was blocked in the amorphic mutant larvae, non-imaginal larval cells continued to grow and endoreplicate their DNA. The Dm cdc2 mutant phenotype could neither be rescued with Dm cdc2c (encoding a cdc2-like kinase) nor enhanced by a reduction of the Dm cdc2c gene dose. These results indicate that the Dm cdc2- and Dm cdc2c-kinases control different processes. PMID- 8223249 TI - Type VI collagen expression is upregulated in the early events of chondrocyte differentiation. AB - Dedifferentiated chondrocytes cultured adherent to the substratum proliferate and synthesize large amounts of type I collagen but when transferred to suspension culture they decrease proliferation, resume the chondrogenic phenotype and the synthesis of type II collagen, and continue their maturation to hypertrophic chondrocyte (Castagnola et al., 1986, J. Cell Biol. 102, 2310-2317). In this report, we describe the developmentally regulated expression of type VI collagen in vitro in differentiating avian chondrocytes. Type VI collagen mRNA is barely detectable in dedifferentiated chondrocytes as long as the attachment to the substratum is maintained, but increases very rapidly upon passage of the cells into suspension culture reaching a peak after 48 hours and declining after 5-6 days of suspension culture. The first evidence of a rise in the mRNA steady-state levels is obtained already at 6 hours for the alpha 3(VI) chain. Immunoprecipitation of metabolically labeled cells with type VI collagen antibodies reveals that the early mRNA rise is paralleled by an increased secretion of type VI collagen in cell media. Induction of type VI collagen is not the consequence of trypsin treatment of dedifferentiated cells since exposure to the actin-disrupting drug cytochalasin or detachment of the cells by mechanical procedures has similar effects. In 13-day-old chicken embryo tibiae, where the full spectrum of the chondrogenic differentiation process is represented, expression of type VI collagen is restricted to the articular cartilage where chondrocytes developmental stage is comparable to stage I (high levels of type II collagen expression).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223250 TI - Capacity to form choroid plexus-like cells in vitro is restricted to specific regions of the mouse neural ectoderm. AB - Neural ectoderm was dissected from 9.5-day and 8.5-day gestation mouse embryos and divided into forebrain, midbrain, hindbrain and spinal cord regions. Forebrain and hindbrain material from 9.5-day neural ectoderm was further divided into presumptive choroid plexus regions and regions that would normally form nervous tissue in vivo. All tissues were plated onto a basement membrane substratum for culture in vitro. It was found that explants of neural ectoderm that would normally form choroid plexus in vivo, readily differentiated to form choroid plexus-like cells in culture. Cells from hindbrain segments and forebrain regions, which would normally form nervous tissue, also had the potential to differentiate into cells resembling the choroid plexus epithelium in culture, provided that the normal cell-cell interactions were disrupted. Cells from the midbrain neuromeres of 9.5-day embryos, which do not form a choroid plexus in vivo, did not form this lineage in vitro. However, cells cultured from the earlier head-fold stage midbrain neural ectoderm could develop into choroid plexus epithelium. There was no evidence that neural ectoderm from the spinal cord had the developmental potential to form choroid plexus epithelial cells at either of these two developmental stages. These studies show that the restrictions in the potential of neural ectoderm stem cells to form different lineages proceeds according to morphological divisions that appear along the anterior-posterior axis during the early stages of brain development. These results suggest that the division of neural ectoderm into segments which contain discrete stem cell populations may be a general feature of the early phase of development of the central nervous system. PMID- 8223251 TI - Mesonephric contribution to testis differentiation in the fetal mouse. AB - Testes from 11.5-day-old mouse embryos, with and without attached mesonephroi, were cultured for 7 days. Isolated testes failed to develop well-differentiated testis cords: however, when cultured attached to a mesonephros from either a male or a female donor embryo, testes developed cords that were normal in appearance. Testes cultured next to a mesonephric region but separated from it by a permeable filter, did not develop normal cords, nor did testes grafted to fragments of embryonic limb or heart. When testes were grafted to mesonephric regions from mice carrying a transgenic marker, the marker was found in some of the peritubular myoid cells and other interstitial cells of the testis, but not in the Sertoli cells or the germ cells. We conclude that after 11.5 days post coitum, cells can migrate from the mesonephric region into the differentiating testis and can contribute to the interstitial cell population, and that this contribution is necessary for the establishment of normal cord structure. The germ cells in all cultured testes, whether or not differentiated cords were present, were T1 prospermatogonia: no meiotic germ cells were seen. PMID- 8223252 TI - Regulation of wingless transcription in the Drosophila embryo. AB - The segment polarity gene wingless (wg) is expressed in a complex pattern during embryogenesis suggesting that it plays multiple roles in the development of the embryo. The best characterized of these is its role in cell pattening in each parasegment, a process that requires the activity of other segment polarity genes including patched (ptc) and hedgehog (hh). Here we present further evidence that ptc and hh encode components of a signal transduction pathway that regulate the expression of wg transcription following its activation by pair-rule genes. We also show that most other aspects of wg expression are independent of this regulatory network. PMID- 8223253 TI - Programmed cell death during Drosophila embryogenesis. AB - The deliberate and orderly removal of cells by programmed cell death is a common phenomenon during the development of metazoan animals. We have examined the distribution and ultrastructural appearance of cell deaths that occur during embryogenesis in Drosophila melanogaster. A large number of cells die during embryonic development in Drosophila. These cells display ultrastructural features that resemble apoptosis observed in vertebrate systems, including nuclear condensation, fragmentation and engulfment by macrophages. Programmed cell deaths can be rapidly and reliably visualized in living wild-type and mutant Drosophila embryos using the vital dyes acridine orange or nile blue. Acridine orange appears to selectively stain apoptotic forms of death in these preparations, since cells undergoing necrotic deaths were not significantly labelled. Likewise, toluidine blue staining of fixed tissues resulted in highly specific labelling of apoptotic cells, indicating that apoptosis leads to specific biochemical changes responsible for the selective affinity to these dyes. Cell death begins at stage 11 (approximately 7 hours) of embryogenesis and thereafter becomes widespread, affecting many different tissues and regions of the embryo. Although the distribution of dying cells changes drastically over time, the overall pattern of cell death is highly reproducible for any given developmental stage. Detailed analysis of cell death in the central nervous system of stage 16 embryos (13-16 hours) revealed asymmetries in the exact number and position of dying cells on either side of the midline, suggesting that the decision to die may not be strictly predetermined at this stage. This work provides the basis for further molecular genetic studies on the control and execution of programmed cell death in Drosophila. PMID- 8223254 TI - A different type of amphibian mesoderm morphogenesis in Ceratophrys ornata. AB - Ceratophrys ornata, the Argentinean horned frog, has a significantly different pattern of early morphogenesis than does the most studied amphibian, Xenopus laevis. Time-lapse videomicroscopy, scanning electron microscopy, histological sections and lineage tracers have shown that, in C. ornata, some prospective notochord, somite and tailbud mesoderm cells leave the surface epithelium of the archenteron by ingression. After gastrulation, SEM reveals cells with constricted apices and a bottle shape in three zones on the archenteron roof and in a fourth zone around the blastopore. Prospective somitic tissue ingresses first from two lateral zones, followed by ingression of prospective notochord from the medial zone and tailbud mesoderm from the circumblastoporal zone. This is unlike X. laevis, in which no cells with constricted apices are present on the dorsal surface of the archenteron, nor do any cells ingress into the deep mesodermal layers from the surface layer. PMID- 8223255 TI - Ultrastructural analysis of Drosophila ovarian follicles differing in yolk polypeptide (yps) composition. AB - Drosophila ovarian follicles were examined ultrastructurally to study the vesicular traffic in the cortical ooplasm. The endocytic pathway leading to the production of yolk spheres was visualized following in vivo or in vitro exposure to peroxidase. The Golgi apparatus and the yolk spheres of wild-type ovarian follicles were preferentially labelled by fixation with osmium zinc iodide (OZI). Labelling of wild-type ovarian follicles was compared to that of several mutant follicles--L186/Basc, fs(2)A17 and ap4--which are defective in vitellogenesis. In these mutants, the Golgi apparatus and the vesicles nearby were either scantly labelled or not labelled at all. In oocytes from flies homozygous for the gene fs(1)1163, the Golgi apparatus was labelled as in the controls, but no yolk spheres appeared to be labelled with OZI at any of the developmental stages. In several Drosophila strains, the pattern of OZI label in the cortical ooplasm was seen to vary in relation to the number of yp structural genes. In starved Drosophila females, OZI labelling of the cortical ooplasm appeared restricted to the Golgi apparatus and to an extended tubular network. A similar labelling pattern was also detected in in vitro cultured vitellogenic follicles. Refeeding, topical application of juvenile hormone analogue to starved females or hormone addition to the culture medium, all caused the yolk spheres to become labelled with OZI and to incorporate peroxidase. These observations prove that impairing endocytic uptake by either mutation or lack of juvenile hormone prevents fusion of coated vesicles and tubules with the yolk spheres and leads them instead to form an intermediate cell compartment with Golgi-derived vesicles. PMID- 8223256 TI - Cellular patterning of fast and slow fibres in the intermandibularis muscle of chick embryos. AB - The way in which the pattern of cell types arises during development of individual muscles was explored. The pattern of cellular differentiation resulting from the synthesis of particular fast and slow myosin heavy chains (MyHC) was investigated in the intermandibularis muscle in the lower jaw of chick embryos. The intermandibularis muscle has a proximodistal pattern of fibre type distribution. The distal region of the muscle contains a ratio of 1.5:1 fast to slow muscle fibres, which increases to > 2.5:1 in the proximal region. The intermandibularis muscle is assembled in a proximodistal sequence, with both fast and slow muscle cells differentiating within the earliest muscle and then establishing the specific pattern of cell types. This pattern is not dependent on a specific innervation source, as normal lower jaw muscles develop and the intermandibularis has the same graded cellular pattern when the mandibular primordium is grafted to the limb bud stump. Micromass cultures were used to explore the pool of potentially myogenic cells that are available to construct the muscles. Even before the muscle differentiates in vivo, both fast and slow cells are present in the primordia. These potentially myogenic cells are already distributed within the primordium in a proximodistal fashion that mimics the cellular pattern found in the muscle that develops. PMID- 8223257 TI - A Y-chromosomal effect on blastocyst cell number in mice. AB - Karyotopic and cell number analysis of 3.5 day post coitum preimplantation mouse embryos was used to determine whether XY embryos had more cells than XX embryos at the late morula/early blastocyst stage. This proved to be the case for the CD1 strain (for which it had previously been shown that XY embryos form a blastocoel earlier than XX embryos) and for the MF1 strain. However, this increased cell number was not seen in MF1 embryos carrying an RIII strain Y in place of the MF1 Y. Furthermore, interstrain crosses between CD1 and the MF1,YRIII strain showed that the cell number increase segregated with the CD1 Y but not with the RIII Y. It is concluded that the CD1 and MF1 Y chromosomes carry a factor that accelerates the rate of preimplantation development. PMID- 8223258 TI - Mutations of the fizzy locus cause metaphase arrest in Drosophila melanogaster embryos. AB - We describe the effects of mutations in the fizzy gene of Drosophila melanogaster and show that fizzy mutations cause cells in mitosis to arrest at metaphase. We show that maternally supplied fizzy activity is required for normal nuclear division in the preblastoderm embryo and, during later embryogenesis, that zygotic fizzy activity is required for the development of the ventrally derived epidermis and the central and peripheral nervous systems. In fizzy embryos, dividing cells in these tissues arrest at metaphase, fail to differentiate and ultimately die. In the ventral epidermis, if cells are prevented from entering mitosis by using a string mutation, cell death is prevented and the ability to differentiate ventral epidermis is restored in fizzy; string double mutant embryos. These results demonstrate that fizzy is a cell cycle mutation and that the normal function of the fizzy gene is required for dividing cells to exit metaphase and complete mitosis. PMID- 8223259 TI - A mRNA localized to the vegetal cortex of Xenopus oocytes encodes a protein with a nanos-like zinc finger domain. AB - mRNAs concentrated in specific regions of the oocyte have been found to encode determinants that specify cell fate. We show that an intermediate filament fraction isolated from Xenopus stage VI oocytes specifically contains, in addition to Vg1 RNA, a new localized mRNA, Xcat-2. Like Vg1, Xcat-2 is found in the vegetal cortical region, is inherited by the vegetal blasomeres during development, and is degraded very early in development. Sequence analysis suggests that Xcat-2 encodes a protein that belongs to the CCHC RNA-binding family of zinc finger proteins. Interestingly, the closest known relative to Xcat 2 in this family is nanos, an RNA localized to the posterior pole of the Drosophila oocyte whose protein product suppresses the translation of the transcription factor hunchback. The localized and maternally restricted expression of Xcat-2 RNA suggests a role for its protein in setting up regional differences in gene expression that occur early in development. PMID- 8223260 TI - Perpendicular contact guidance of CNS neuroblasts on artificial microstructures. AB - Rodent CNS neuroblasts show parallel and perpendicular contact guidance behaviors on aligned neurite bundles in microexplant cultures (Nakatsuji, N. and Nagata, I. (1989) Development, 106, 441-447; N. I. and N. N. (1991) ibid., 112, 581-590). To test the hypothesis that the physical surface structure of the neurite bundle causes the perpendicular contact guidance, we cultured dissociated neuroblasts on quartz plates on which grating-like microstructures were fabricated by lithographic techniques. Various types of CNS neuroblasts, but not PNS neurons, oriented their processes and migrated both perpendicular and parallel to the axis of the microstructure. Perpendicular orientation was frequently observed when the microstructured grooves had depths between 0.3 micron and 0.8 micron and a width of 1 micron, which roughly mimics a tightly aligned neurite bundle. Thus, CNS neuroblasts have the ability to extend their processes and migrate perpendicular to aligned surface microstructures. PMID- 8223261 TI - Repression of Drosophila pair-rule segmentation genes by ectopic expression of tramtrack. AB - The tramtrack (ttk) protein has been proposed as a maternally provided repressor of the fushi tarazu (ftz) gene in Drosophila embryos at the preblastoderm stage. Consistent with this hypothesis, we have detected by immunohistochemistry the presence of ttk protein in preblastoderm embryos. This is followed by a complete decay upon formation of the cellular blastoderm when ftz striped expression is at its peak. In addition, the highly complex pattern of zygotic ttk expression suggests specific functions for ttk late in development that are separate from the regulation of ftz. We have produced ttk protein ectopically in blastoderm stage embryos transformed with a heat shock-ttk construct. Ectopic ttk caused complete or near-complete repression of the endogenous ftz gene, as well as significant repression of the pair-rule genes even skipped, odd skipped, hairy and runt. These findings suggest that specific repression by ttk (or by undiscovered repressors) may be more than an isolated phenomenon during the rapid cleavage divisions, a period when the need for genetic repression has not been generally anticipated. PMID- 8223262 TI - Functional and molecular characterization of the transcriptional regulatory region of Tcp-10bt, a testes-expressed gene from the t complex responder locus. AB - Mouse t haplotypes contain several mutant alleles that disrupt spermatogenesis. Their phenotypes include sterility, reduced fertility and transmission ratio distortion (TRD). The substantial genetic analyses of these mutant alleles, coupled with intensive physical characterization of the t complex, provides a fertile ground for identifying and understanding genes essential to male gametogenesis. The t complex responder (Tcr) locus plays a central role in this process, interacting with other t haplotype-encoded genes to mediate TRD. A candidate responder gene, Tcp-10bt, has been cloned and subjected to molecular characterization. Here, we define the transcriptional regulatory regions of this gene in transgenic mice. A 1.6 kb (but not 0.6 kb) DNA fragment upstream of the transcription start site contains all the regulatory signals for appropriate temporal and germ cell-specific expression of this gene. Two smaller fragments within this region bound specifically to nuclear factor(s) from germ cell protein extracts in gel shift assays. This work is a step towards understanding the mechanism of Tcp-10bt regulated expression and may ultimately help reveal a common regulatory pathway shared by other similarly expressed spermatogenic genes. PMID- 8223263 TI - Changing patterns of gene expression define four stages of cerebellar granule neuron differentiation. AB - Among CNS neuronal populations, the cerebellar granule cell provides a simple model for analysing the molecular regulation of CNS neurogenesis. In this study, polyclonal antisera raised against immature granule cell precursors, purified from early postnatal mouse cerebellum, were used to isolate 39 unique cDNA clones from a lambda gt11 cDNA expression library made from the same cell population. Northern blot analysis revealed developmental stage and tissue-specific expression of 28 of the clones. In situ localization of mRNAs encoded by these novel cDNAs, as well as those encoding the axonal glycoprotein TAG-1 and the alpha 6 subunit of the GABAA receptor, reveal four distinct stages in cerebellar granule cell differentiation. The developmentally transient and spatially restricted expression of clones GC9 and GC44 identify a previously unrecognized step in cerebellar histogenesis. PMID- 8223264 TI - Multiple cell interactions are required for fate specification during male spicule development in Caenorhabditis elegans. AB - The B blast cell divides postembryonically in C. elegans males to produce 47 progeny that include all of the cells of the copulatory spicules. During the early development of the B lineage, the anterior daughter of B, B.a, generates eight cells. These cells migrate to form four pairs of cells that flank the developing cloaca (ventral, dorsal, and two identical lateral pairs). For each pair, the more anterior cell produces a distinct lineage ('anterior fate') from the posterior cell ('posterior fate'). For the ventral and dorsal pairs, either cell can migrate to the anterior position and produce the anterior lineage, and the other cell migrates posterior and produces the posterior lineage (Sulston and Horvitz, 1977, Dev. Biol. 56, 110-156). The migration is variable, although the resultant fate pattern is invariant. In the two lateral pairs, both the migration and fate pattern are invariant. Using a laser microbeam to selectively ablate neighboring cells we have found that the cells of the lateral pair also respond to positional cues. For all four pairs other male-specific blast cells provide extracellular cues. In general, F and U promote anterior fates, Y promotes some posterior fates, and the B.a progeny promote posterior fates. Several of these cues are redundant. By ablating combinations of cells we have deduced how these signals may act in concert to specify the fates of the B.a progeny. We propose that fate specification in these pairs depends on three general classes of extracellular cues: positional cues, modulators of positional cues, and lateral signals. The B lineage thus provides an opportunity to study with single cell resolution the integration of multiple intercellular signals. PMID- 8223265 TI - Genetic analysis of laminin A reveals diverse functions during morphogenesis in Drosophila. AB - In order to dissect the functions of laminin A in vivo, we have undertaken a molecular and genetic characterization of the laminin A subunit (lamA) gene in Drosophila. Sequence analysis predicts a multidomain structure similar to mammalian homologs. We generated a series of complete and partial loss-of function mutant alleles of the lamA gene; complete loss-of-function mutations lead to late embryonic lethality. Certain combinations of partial loss-of function lamA alleles give rise to escaper adults, which have rough eyes associated with changes in cell fate and pattern, misshapen legs and defects in wing structure. These phenotypes suggest that laminin A has diverse functions during morphogenesis in Drosophila. PMID- 8223266 TI - Two distinct populations of prestalk cells within the tip of the migratory Dictyostelium slug with differing fates at culmination. AB - The ecmA gene of Dictyostelium encodes an extracellular matrix protein and is selectively expressed in prestalk cells. We show that its promoter contains discrete elements that direct expression in different subpopulations of prestalk cells. Prestalk(pst)A cells occupy the front half of the prestalk region. Expression in pstA cells requires DNA sequences close to the cap site of the gene and a separate, upstream region that acts in combination with the gene proximal sequences. PstO cells are situated in the rear half of the prestalk region and at least two separate and redundant promoter regions direct expression within them. All constructs that are expressed in pstO cells are also expressed in anterior like cells (ALCs); cells that resemble prestalk cells but which are scattered throughout the prespore region. This observation suggests that pstO cells and ALCs may be very similar in their properties. If development occurs under conditions in which a migratory slug is not formed, there is an ordered movement of cells into the stalk tube. PstA cells enter the stalk tube first, followed by a proportion of the pstO cells. The remainder of the pstO cells contribute to the upper cup, an ALC-derived subpopulation of prestalk cells which is located at the apex of the spore head. After prolonged slug migration, a discrete pstO zone appears not to be maintained and, at culmination, pstO cells are found scattered throughout the stalk. PMID- 8223267 TI - Function and spatial distribution in developing chick retina of the laminin receptor alpha 6 beta 1 and its isoforms. AB - We have recently shown that the laminin-binding integrin receptor, alpha 6 beta 1, is prominently expressed in the developing chick retina, and its expression and activity are regulated during development on both retinal ganglion cells and other neural retinal cells. In the present study, we show that antibodies specific for the extracellular portion of the chick alpha 6 subunit dramatically inhibit interactions in vitro between embryonic day 6 neural retinal cells and laminin, showing that alpha 6 beta 1 functions as an important laminin receptor on developing retinal neurons. In previous work, we showed that alpha 6 mRNA levels on retinal ganglion cells decrease dramatically after E6 during the period that RGC axons innervate the optic tectum. In the present study, we show decreases in alpha 6 mRNA are not prevented by ablation of the optic tectum, indicating that tectal contact is not the major cause of this decrease. Within the embryonic retina, the alpha 6 subunit is codistributed, in part, with laminin, suggesting that it functions as a laminin receptor during retina development in vivo. Furthermore, two isoforms of the alpha 6 protein with distinct cytoplasmic domains generated by differential splicing have quite different distribution patterns in the retina, suggesting that these two isoforms may have different functions during retinal development. PMID- 8223268 TI - Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. AB - We have designed a system for targeted gene expression that allows the selective activation of any cloned gene in a wide variety of tissue- and cell-specific patterns. The gene encoding the yeast transcriptional activator GAL4 is inserted randomly into the Drosophila genome to drive GAL4 expression from one of a diverse array of genomic enhancers. It is then possible to introduce a gene containing GAL4 binding sites within its promoter, to activate it in those cells where GAL4 is expressed, and to observe the effect of this directed misexpression on development. We have used GAL4-directed transcription to expand the domain of embryonic expression of the homeobox protein even-skipped. We show that even skipped represses wingless and transforms cells that would normally secrete naked cuticle into denticle secreting cells. The GAL4 system can thus be used to study regulatory interactions during embryonic development. In adults, targeted expression can be used to generate dominant phenotypes for use in genetic screens. We have directed expression of an activated form of the Dras2 protein, resulting in dominant eye and wing defects that can be used in screens to identify other members of the Dras2 signal transduction pathway. PMID- 8223269 TI - Zebrafish primary neurons initiate expression of the LIM homeodomain protein Isl 1 at the end of gastrulation. AB - Isl-1 has previously been established as the earliest marker of developing chicken spinal motor neurons where it is regulated by inductive signals from the floorplate and notochord. We now report that, in zebrafish, the expression of Isl 1 is initiated in Rohon-Beard cells, primary motor neurons, interneurons and cranial ganglia, hours before the neural tube itself is formed. The expression is initiated simultaneously in the Rohon-Beard cells and the primary motor neurons, at the axial level of the presumptive first somite. The Isl-1-expressing motor neurons appear on either side of the ventral midline whereas the interneurons and Rohon-Beard cells initiate expression while located at the edge of the germinal shield. Isl-1 expression is initiated in these cells before the formation of a differentiated notochord. Isl-1 is expressed in the various functional classes of primary neurons at 24 hours postfertilization. This selective expression of a homeodomain protein in the primary neurons implies that these neurons share a common program of early development and that they have evolved and been selected for as a coordinated system. One of the functions of the primary neurons is to send long axons which pioneer the major axon tracts in the zebrafish embryo. An evolutionary conserved functional role for Isl-1 in the expression of the pioneering phenotype of the primary neurons is suggested. PMID- 8223270 TI - wingless expression mediates determination of peripheral nervous system elements in late stages of Drosophila wing disc development. AB - We have used conditional wingless genotypes to dissect the role of this gene in late stages of wing disc development. One of these genotypes (wgIL/wg-lacZ) is simultaneously a reporter of wingless transcription and temperature-sensitive for wingless function, and has allowed us to define its pattern of transcription in the absence of wingless activity. The primordia of a subset of the bristles of the notum, which develop in or immediately adjacent to wingless-expressing cells, depend upon wingless activity. The time-course of this contribution and the effect on proneural gene expression together suggest that wingless may regulate the activity of products of the achaete-scute complex in proneural clusters. wingless activity is also required at the presumptive wing margin and is a necessary precondition for the change in proliferation pattern in this region. The involvement of wingless in transducing or mediating positional signals for spatial patterning in imaginal disc development is discussed. PMID- 8223271 TI - Differential expression of the HMG box factors TCF-1 and LEF-1 during murine embryogenesis. AB - The recent identification of a number of T lymphocyte-specific enhancers has allowed the cloning of several novel transcription factors. Two of these, TCF-1 and LEF-1, contain a virtually identical DNA-binding domain of the High Mobility Group (HMG-1) box type. TCF-1 and LEF-1 originate from a recent gene duplication event as evidenced by comparison with the chicken homologue, chTCF. We have now analyzed the differential expression of these two transcription factors. In a panel of lymphoid cell lines, TCF-1 was exclusively expressed in the T cell lineage. In contrast, LEF-1 mRNA was detected at equivalent levels in pro- and pre-B cells and in all T lineage cells. In situ hybridization on murine embryos revealed that TCF-1 and LEF-1 were widely expressed at day 7.5 of gestation. At later stages, the expression patterns were complex and only partially overlapping. The expression of TCF-1 and LEF-1 coincided until day 10.5, when mRNAs were detected in limb buds, neural crest, pharyngeal arches and nasal process. At later time points (day 13.5 to 14.5), sites of overlapping expression included lung, the urogenital system, tooth buds, thymus and choroid plexus. Unique expression sites for TCF-1 included Reichert's membrane and trophectoderm derived cells, the ribs and thoracic prevertebrae, craniofacial structures, the adrenal gland and meninges. Unique LEF-1 expression was observed in the tail prevertebrae, brain and inner ear. Postnatally, expression of both genes could only be detected in lymphoid tissues. These observations suggest that TCF-1 and LEF-1 exert differential functions during murine embryogenesis. PMID- 8223272 TI - Evolutionary change in neural development within the arthropods: axonogenesis in the embryos of two crustaceans. AB - It has been previously suggested that there is a conservative program for neural development amongst the arthropods, on the basis that a stereotyped set of cells involved in establishing the axon tracts in the CNS of insect embryos is also present in crayfish embryos. We have examined the spatiotemporal pattern of axon growth from a set of early differentiating central neurons in the embryo of two crustaceans, the woodlouse Porcellio scaber and the freshwater crayfish Cherax destructor, and drawn comparisons with insect neurons whose somata lie in corresponding positions within the CNS. While many of the woodlouse and crayfish neurons show a similar pattern of axon growth to their insect counterparts, the axon trajectories taken by others differ from those seen in insects. We conclude that this aspect of early neural development has not been rigidly conserved during the evolution of the crustaceans and insects. However, the extent of similarity between the insects and the crustaceans is consistent with the idea that these groups of arthropods share a common evolutionary 'Bauplan' for the construction of their nervous systems. While the pattern of early axon growth in the woodlouse and crayfish embryos is sufficiently similar that many neurons could be confidently recognised as homologues, several differences were noted in both the relative order of axon outgrowth and axon morphologies of individual neurons. PMID- 8223273 TI - trkC, a receptor for neurotrophin-3, is widely expressed in the developing nervous system and in non-neuronal tissues. AB - The Trk family of tyrosine kinases encodes receptors for nerve growth factor related neurotrophins. Here we present a developmental expression study of trkC, which encodes a receptor for neurotrophin-3 (NT-3). Like the related genes, trk and trkB, trkC is expressed primarily in neural lineages although the pattern is complex and includes non-neuronal cells. Direct comparison with trk and trkB developmental expression patterns permits the following observations. (1) trkC is expressed in novel neural tissues where other Trk genes are silent. (2) Some tissues appear to coexpress trkB and trkC receptors in the embryo and in the adult. (3) trkC expression can be detected in the gastrulating embryo. These data provide insights into the role of Trk-family receptors and nerve growth factor related neurotrophins during development and suggest that, in addition to regulating neuronal survival and differentiation, the neurotrophin/Trk receptor system may have broader physiological effects. Finally, interspecific mouse backcrosses have been used to map the location of each of the Trk genes on mouse chromosomes. Alignment with available chromosomal maps identify possible linkage between the Trk genes and known neurological mutations. PMID- 8223274 TI - FGF signalling in the early specification of mesoderm in Xenopus. AB - We have examined the role of FGF signalling in the development of muscle and notochord and in the expression of early mesodermal markers in Xenopus embryos. Disruption of the FGF signalling pathway by expression of a dominant negative construct of the FGF receptor (XFD) generally results in gastrulation defects that are later evident in the formation of the trunk and tail, though head structures are formed nearly normally. These defects are reflected in the loss of notochord and muscle. Even in embryos that show mild defects and gastrulate properly, muscle formation is impaired, suggesting that morphogenesis and tissue differentiation each depend on FGF. The XFD protein inhibits the expression of the immediate early gene brachyury throughout the marginal zone, including the dorsal side; it does not, however, inhibit the dorsal lip marker goosecoid, which is expressed in the first involuting mesoderm at the dorsal side that will underlie the head. The XFD protein also inhibits Xpo expression, an immediate early marker of ventral and lateral mesoderm. These results suggest that FGF is involved in the earliest events of most mesoderm induction that occur before gastrulation and that the early dorsal mesoderm is already composed of two cell populations that differ in their requirements for FGF. PMID- 8223275 TI - flk-1, an flt-related receptor tyrosine kinase is an early marker for endothelial cell precursors. AB - We have used RT-PCR to screen pluripotent murine embryonic stem cells to identify receptor tyrosine kinases (RTKs) potentially involved in the determination or differentiation of cell lineages during early mouse development. Fourteen different tyrosine kinase sequences were identified. The expression patterns of four RTKs have been examined and all are expressed in the mouse embryo during, or shortly after, gastrulation. We report here the detailed expression pattern of one such RTK, the flt-related gene flk-1. In situ hybridization analysis of the late primitive streak stage embryo revealed that flk-1 was expressed in the proximal-lateral embryonic mesoderm; tissue fated to become heart. By headfold stages, staining was confined to the endocardial cells of the heart primordia as well as to the blood islands of the visceral yolk sac and the developing allantois. Patchy, speckled staining was detected in the endothelium of all the major embryonic and extraembryonic blood vessels as they formed. During early organogenesis, expression was detected in the blood vessels of highly vascularized tissues such as the brain, liver, lungs and placenta. Since flk-1 was expressed in early mesodermal cells prior to any morphological evidence for endothelial cell differentiation (vasculogenesis), as well as in cells that form blood vessels from preexisting ones (angiogenesis), it appears to be a very early marker of endothelial cell precursors. We have previously reported that another novel RTK, designated tek, was expressed in differentiating endothelial cells. We show here that flk-1 transcripts are expressed one full embryonic day earlier than the first tek transcripts. The expression of these two RTKs appear to correlate with the specification and early differentiation of the endothelial cell lineage respectively, and therefore may play important roles in the establishment of this lineage. PMID- 8223276 TI - Lachesin: an immunoglobulin superfamily protein whose expression correlates with neurogenesis in grasshopper embryos. AB - We describe the developmental expression in grasshopper (Schistocerca americana) and molecular characterization in grasshopper and fruit fly (Drosophila melanogaster) of Lachesin, a novel immunoglobulin superfamily protein. Lachesin is expressed on the surfaces of differentiating neuronal cells from the onset of neurogenesis in both the central and peripheral nervous systems. Lachesin expression begins in some cells of the neurogenic ectoderm immediately after engrailed expression begins in the posterior cells of each future segment. All neurogenic cells express Lachesin early, but only those cells that become neuroblasts continue to express Lachesin. Ectodermal cells in the neurogenic region that adopt non-neuronal fates lose Lachesin at the time that they diverge from a potentially neurogenic pathway. Neuroblasts, ganglion mother cells and neurons all express Lachesin early in their lives, but expression becomes restricted to a subset of neurons as development progresses. Sensory neurons express Lachesin as they delaminate from the body wall ectoderm. Lachesin is also present on growing axons of the CNS and PNS and becomes restricted to a subset of axons later in development. This expression is unique among known insect neurogenic genes and suggests a role for Lachesin in early neuronal differentiation and axon outgrowth. Grasshopper Lachesin is a 38 x 10(3) M(r) protein linked to cell membranes through a glycosyl phosphatidylinositol anchor. We have cloned the Lachesin gene from both grasshopper and fly. The proteins are highly conserved (70% identical) between the two species. Lachesin is similar to Drosophila amalgam, bovine OBCAM and the human poliovirus receptor, putting it into a subgroup of the immunoglobulin superfamily containing one V- and two C2 type immunoglobulin domains. Lachesin is also similar to several other vertebrate immunoglobulin superfamily proteins (TAG-1, F11, L1 and NgCAM) known to function in neurite outgrowth and other cell surface recognition events. PMID- 8223277 TI - Selective dispersal of avian rhombomere cells in orthotopic and heterotopic grafts. AB - During hindbrain development, cells become segregated into segmental groups, rhombomeres, by mechanisms that are presently unknown. One contributory factor early in development may be an alternating periodicity in cell surface properties down the neuraxis. This possibility was previously suggested by experiments in which tissue from different segmental levels was apposed in the absence of a boundary. New boundaries were regenerated only when rhombomeres from adjacent positions or positions three rhombomeres distant from one another were apposed. Combinations of two odd-numbered or two even-numbered rhombomeres usually failed to generate a boundary. In order to pursue this phenomenon to the cellular level, we have used two approaches, both involving donor-to-host transplantation. First, quail rhombomeres were grafted at various hindbrain levels of a chick host. Apposition of rhombomere 4 (r4) with r3 was concomitant with negligible cell mixing across the interface. By contrast, combinations of r3 with r5 or with r3 tissue led to cell mixing that was more extensive in combinations of identical rhombomeres (r3 with r3) than between two alternate ones (r3 with r5). Secondly, we grafted small pieces of fluorescently prelabelled chick rhombomere tissue at various hindbrain levels of chick hosts. In most cases, cells dispersed widely when transplanted orthopically or two segments distant from that of their origin. Cells transplanted into an adjacent segment, however, showed a tendency to remain undispersed. Among the different graft combinations, furthermore, there was a variation in the extent of dispersal that showed an additional level of complexity not revealed in boundary regeneration experiments. The possibility is raised that the early partitioning of rhombomeres involves a hierarchy in the adhesive preferences of cell-cell interactions along the neuraxis. PMID- 8223278 TI - PDGF and its receptors in the developing rodent retina and optic nerve. AB - We have used in situ hybridization to visualize cells in the developing rat retina and optic nerve that express mRNAs encoding the A and B chains of platelet derived growth factor (PDGF-A and PDGF-B), and the alpha and beta subunits of the PDGF receptor (PDGF-alpha R and PDGF-beta R). We have also visualized PDGF-A protein in these tissues by immunohistochemistry. In the retina, PDGF-A mRNA is present in pigment epithelial cells, ganglion neurons and a subset of amacrine neurons. PDGF-A transcripts accumulate in ganglion neurons during target innervation and in amacrine neurons around the time of eye opening, suggesting that PDGF-A expression in these cells may be regulated by target-derived signals or by electrical activity. In the mouse retina, PDGF-A immunoreactivity is present in the cell bodies, dendrites and proximal axons of ganglion neurons, and throughout the inner nuclear layer. PDGF-alpha R mRNA is expressed in the retina by astrocytes in the optic fibre layer and by a subset of cells in the inner nuclear layer that might be Muller glia or bipolar neurons. Taken together, our data suggest short-range paracrine interactions between PDGF-A and PDGF-alpha R, the ligand and its receptor being expressed in neighbouring layers of cells in the retina. In the optic nerve, PDGF-A immunoreactivity is present in astrocytes but apparently not in the retinal ganglion cell axons. PDGF-alpha R+ cells in the optic nerve first appear near the optic chiasm and subsequently spread to the retinal end of the nerve; these PDGF-alpha R+ cells are probably oligodendrocyte precursors (Pringle et al., 1992). RNA transcripts encoding PDGF-B and PDGF-beta R are expressed by cells of the hyaloid and mature vascular systems in the eye and optic nerve. PMID- 8223279 TI - Oligodendrocyte precursors originate at the ventral ventricular zone dorsal to the ventral midline region in the embryonic rat spinal cord. AB - The precursors for oligodendrocytes, the myelinating cells of the vertebrate CNS, appear to be initially restricted to ventral regions of the embryonic rat spinal cord. These cells subsequently populate dorsal spinal cord regions where they acquire the mature characteristics of oligodendrocytes. To determine the location and timing of proliferation of oligodendrocyte precursors in the ventral spinal cord, and to map their pathways of migration in vivo, an assay that identifies mitotic cells was used in conjunction with antibodies that distinguish astrocytes, oligodendrocytes and their precursors. Between E16.5 and E18.5, two hours after a maternal injection of BrdU, the majority of proliferating cells were located in a discrete cluster at the ventral ventricular zone dorsal to the ventral midline region of the developing spinal cord. By contrast, 12-24 hours following a BrdU injection at E16.5, increasing numbers of labeled cells were seen in the dorsal and more lateral locations of the spinal cord. These observations suggest that BrdU-labeled ventral ventricular cells, or their progeny migrate dorsally and laterally during subsequent spinal cord development. To determine the nature of these proliferating cells, cultures of dorsal and ventral spinal cord from BrdU-labeled animals were double-labeled with antibodies that identify oligodendrocytes or astrocytes and anti-BrdU. In dorsal spinal cord cultures derived from animals that had received a single injection of BrdU at E16.5, the majority of proliferating cells differentiated into astrocytes while, in ventrally derived cultures from the same animals, the majority of proliferating cells differentiated into oligodendrocytes. In dorsal cultures prepared from animals that received multiple injections of BrdU between E16.5 and E18.5, many more cells were labeled with BrdU and approximately half of these differentiated into oligodendrocytes. These observations suggest that during embryonic development proliferating oligodendrocyte precursors are initially located at the ventral ventricular zone dorsal to the ventral midline region of the spinal cord and during subsequent maturation these cells or their progeny migrated dorsally in the ventricular region of the spinal cord, and laterally to reside in the developing white matter. PMID- 8223280 TI - Expression of the Fgf6 gene is restricted to developing skeletal muscle in the mouse embryo. AB - Fgf6, a member of the Fibroblast Growth Factor (FGF) family, is developmentally regulated and its expression is highly restricted in the adult. To gain further insight into the role of Fgf6, we studied its expression during embryogenesis using RNA in situ hybridization. Fgf6 expression is restricted to developing skeletal muscle. Fgf6 transcripts are first detected in the somites at 9.5 days post-conceptus, and expression continues in developing skeletal muscles up to at least 16.5 days post-conceptus. Fgfr4 is a putative receptor for FGF6. Its pattern of expression during myogenesis overlaps that of Fgf6, but both genes are not expressed in exactly the same population of cells. In addition, recombinant FGF6 protein is able to repress the terminal differentiation of myoblasts in culture, providing additional support to the concept that FGF6 plays an important role in myogenesis. PMID- 8223281 TI - Puffs and PCR: the in vivo dynamics of early gene expression during ecdysone responses in Drosophila. AB - The steroid hormone ecdysone orchestrates insect development by regulating gene networks. In Drosophila the most detailed description of ecdysone action is the sequential activation of early and late puffs in the polytene chromosomes of the late larval salivary gland. A number of these early puffs (2B5, 74EF and 75B) contain complex transcription units (Broad-Complex, E74 and E75 respectively) encoding families of regulatory proteins which are expressed in most if not all tissues. In vitro, transcripts of the different isoforms of these early genes as well as the ecdysone receptor (EcR) present varying dose response characteristics (Karim and Thummel, 1992, EMBO J. 11, 4083-4093). We have developed an in vivo approach using a reverse transcription-polymerase chain reaction assay (RT-PCR) so as to visualise these transcripts in the RNA extracted from a single salivary gland. Using one salivary gland lobe for developmental puff staging and the sister lobe for RT-PCR, we have obtained precise developmental profiles for these transcripts and have extended our study to other tissues and stages where puffing studies were not possible. In the salivary gland we have characterised three distinct ecdysone responses. For the mid and late third larval instar responses our results confirm and extend the conclusions of the in vitro studies concerning the temporal expression of the early gene isoforms. The relatively brief prepupal response contains elements in common with each of the larval responses and all three can be explained by the profiles of the respective ecdysone peaks. Interestingly EcR transcripts respond differently during each response. The analysis of different tissues of the same animal reveals subtle differences in the timing of the ecdysone response and isoform expression and suggests that this may reflect tissue differences in the ecdysone profiles. As these molecules have homologues in vertebrates, our analysis may have general implications for the organisation of hormonal responses in vivo. PMID- 8223282 TI - Catenins in Xenopus embryogenesis and their relation to the cadherin-mediated cell-cell adhesion system. AB - In the course of an analysis of cell-cell adhesion in the Xenopus embryo, antibodies directed against alpha- and beta-catenin were applied to investigate their relation to the cadherins occurring early in this system. The results demonstrate that alpha- and beta-catenin are provided maternally and increase in amount throughout embryogenesis. Immunoprecipitations indicate that both of the catenins are complexed to U-cadherin in the early phase of embryogenesis and to E cadherin, when it appears during gastrulation. An excess of alpha-catenin occurs in free form in the early embryo, whereas all of the beta-catenin seems to be complexed to cadherin. Synthesis of the two components throughout early embryogenesis and their binding to newly synthesized cadherins were demonstrated by metabolic labelling. The spatial distribution of alpha-catenin was analysed by immunohistology. During cleavage alpha-catenin is deposited evenly along the plasma membranes within the embryo, while the cell peripheries at the surface of the embryo remain devoid of alpha-catenin. At later stages, the pattern of alpha catenin distribution becomes more complex. Quantitative differences in the intensity of staining along the plasma membranes in the different regions of the embryo can be distinguished. Particularly the appearance of E-cadherin in the gastrula ectoderm is accompanied by conspicuous depositions of alpha-catenin along the respective plasma membranes in this layer. All cells in the later embryo, apart from the neural crest cells, carry alpha-catenin on their plasma membranes indicating the universal character of cadherin-mediated cell-cell adhesion in the Xenopus embryo. PMID- 8223283 TI - Inactivation of an X-linked transgene in murine extraembryonic and adult tissues. AB - Transgenes located on the X chromosome have been used to study the mechanisms involved in X-chromosome inactivation. Analysis of the transgenic mouse strain M TKneo1 carrying a neomycin resistance gene inserted in the X chromosome showed that, in adult somatic tissues, this transgene is subject to X-inactivation and to de novo methylation as other endogenous X-linked genes. During mouse embryogenesis, X-linked genes show a preferential paternal inactivation in extraembryonic tissues, whereas these genes are subject to random inactivation in embryonic tissues. It has been suggested that, in the mouse, the extraembryonic tissues carry a parental imprint at the time of inactivation. The study of the neo transgene expression in extraembryonic endoderm has shown not only that neo is inactivated but also that, at the RNA level, paternal inactivation of the transgene seems essentially complete. The differences between our results and previously obtained results with a mouse alpha-fetoprotein transgene, which was only inactivated in neonatal tissues but not in extraembryonic tissues, are discussed. PMID- 8223284 TI - Expression of epithelial alkaline phosphatase in segmentally iterated bands during grasshopper limb morphogenesis. AB - Although the study of rostral-caudal segmentation of the insect body has been a rich source of information about embryonic pattern formation, relatively little is known of the process of proximal-distal segmentation of insect appendages. Here we demonstrate that during the period of limb segmentation, five segmentally iterated, sharply demarcated bands of cell surface alkaline phosphatase activity are expressed in embryonic grasshopper limbs. These bands span each intersegmental boundary in the limb as well as one boundary within the tarsus. Within appendages, expression is restricted to epithelial cells, where activity is present on both apical and basolateral surfaces. This epithelial alkaline phosphatase remains active at neutral pH, is insensitive to levamisole inhibition, and is strongly inhibited by nucleoside monophosphates. Treatment of embryos with phosphatidylinositol-specific phospholipase C releases almost all visible chromogenic activity, indicating that the epithelial alkaline phosphatase is anchored to the plasma membrane by glycosyl-phosphatidylinositol. When material released by phosphatidylinositol-specific phospholipase C is separated on native polyacrylamide gels, a single broad band of enzymatic activity is detected following incubation with substrate. A polyclonal antiserum raised against a 55 x 10(3) M(r) alkaline phosphatase from shrimp recognizes a single band of 56 x 10(3) M(r) on immunoblots of grasshopper membrane proteins. The spatially restricted expression of epithelial alkaline phosphatase suggests that it may be involved in epithelial cell rearrangements or shape changes associated with limb segmentation and morphogenesis. It also may contribute to definition of axon routes in the limb, since pioneer afferent growth cones turn at, and migrate along, the edge of one alkaline phosphatase-expressing epithelial domain. PMID- 8223285 TI - The atelectatic ear. PMID- 8223286 TI - Toxoplasmosis appearing as an anterior neck mass. PMID- 8223287 TI - Giant vocal polyp masquerading as asthma. PMID- 8223288 TI - Undifferentiated carcinoma of the nose and paranasal sinuses. An immunohistochemical and clinical study. AB - Among twenty-six "undifferentiated" tumors of the nasal cavity and paranasal sinuses treated from 1970 to 1990 at the Institute of Otolaryngology of Florence University, 13 were ultimately diagnosed as true undifferentiated sinusonasal carcinoma (SNUC) by conventional light microscopy and use of monoclonal antibodies to epithelial membrane antigen and cytokeratins. SNUC patients, who ranged in age from 20 to 82 years, often had multiple sinonasal symptoms due to very large tumors (nine of 13 tumors were staged as T3-T4) with short average delay of 4 months between onset of symptoms and diagnosis. Both data suggest the high growth capacity and aggressiveness of such a tumor. In our series, follow-up evidenced an overall crude 5-year survival rate of 15.5%. Worse prognostic factors are neck metastases and orbital invasion, according to the behavior of more common carcinomas of the nose and paranasal sinuses. We also found a better prognosis for SNUC primarily arisen in the nasal cavity than in paranasal sinuses (crude 5-year survival rate of 66% vs. 10%, respectively). The histopathological and clinical analysis of our series shows that SNUC is a highly aggressive, uncommon tumor of the nose paranasal sinuses, which should be recognized in advance for a more aggressive treatment by combined multiple therapy. PMID- 8223289 TI - Endoscopic stenting of nasofrontal communication in frontal sinus disease. AB - The volume of literature on frontal sinus surgery suggests that the success of surgery to cure frontal sinus pathology is inversely proportional to the total number of techniques described. In an attempt to develop an effective means of treating acute and chronic frontal sinusitis, including iatrogenic naso frontal communication obstruction, an endoscopic stenting procedure has evolved. This method might be contemplated, in selected patients, as the preferred surgery or a useful low morbidity alternative, to more invasive traditional frontal sinus surgery. A high degree of technical difficulty is associated with endoscopic frontal sinus procedures, and they are definitely not recommended for the novice or occasional sinus surgeon. Several years of follow up are required to evaluate the validity of any frontal sinus procedure. More observation of these subjects' clinical progress is necessary before endoscopic stenting of the nasofrontal communication passes the test of time. PMID- 8223290 TI - Endoscopic view of eustachian tube orifice. PMID- 8223291 TI - Radical removal of a large glossopharyngeal neurinoma with preservation of cranial nerve functions. AB - A 51-year-old man is reported who was suffering from an extensive right-sided glossopharyngeal neurinoma (4.6 x 3.4 cm). The patient was admitted with a history of headache for six to seven years and vomiting for two years accompanied by a progressive hearing loss, tinnitus and dizziness during the last year. Audiometry indicated a perceptive deafness in the whole frequency range up to 70 dB HL, while electronystagmography (ENG) showed a loss of vestibular function of the right side, but there were no signs of a jugular foramen syndrome. Magnetic resonance imaging (MRI) revealed a large tumor portion in the right cerebello pontine angle with only a small part in the jugular foramen. The neurinoma was completely removed by microsurgery through a suboccipital approach with preservation of nerves VII-XII. The postoperative course was uneventful and normal function of facial and caudal cranial nerves (Nn IX-XII) were proven by electromyography and magnetic stimulation, with exception of a transitory hypesthesia in the palatine region which completely normalized within a few months. The right-sided hearing loss was unchanged, but vertigo improved. In comparison with literature review the lack of temporary or permanent postoperative dysfunctions of caudal cranial nerves as well as of the facial nerve was extraordinary in the reported case. PMID- 8223292 TI - Nasal signs and symptoms of the allergic response--seasonal rhinitis. PMID- 8223293 TI - Pediatric actinomycosis of the head and neck. AB - Actinomycosis is a rare soft tissue infection caused by a gram-positive, anaerobic bacteria. It is seen only approximately once a year in major medical centers, and is rarer still in the pediatric population. About 50% of all cases of actinomycosis involve the head and neck. The organism is very difficult to isolate in culture and the differential diagnosis is extensive, prompting one reviewer to name actinomycosis the "masquerader" of the head and neck. We review three cases of actinomycosis over the past ten years (1981-1990) at Texas Children's Hospital, Houston, Texas. PMID- 8223294 TI - Fatal mediastinal abscess from upper respiratory infection. AB - Suppurative mediastinitis following neck infection is an uncommon entity that carries a 40% mortality. It is most commonly associated with esophageal perforation or odontogenic abscess. The rapidity of spread has been attributed to dependent drainage from the neck into the mediastinum, negative intrathoracic pressure, and synergistic necrotizing bacterial growth. Cultures obtained usually grow streptococci and/or Bacteroides. We report the case of a 34-year-old woman with an upper respiratory infection who subsequently was found to have a para /retropharyngeal and mediastinal abscess. Septic shock, respiratory failure, and death ensued, despite aggressive treatment with broad-spectrum antibiotics (for both aerobes and anaerobes), surgical drainage of the neck and mediastinum, and cardiorespiratory support. The danger of a rapid downhill clinical course with mortality due to suppurative mediastinitis makes early diagnosis of critical importance. All clinicians evaluating a "sore throat" should consider neck edema or gas on neck radiograms as evidence of cervical abscess, and subsequent widening of the mediastinum on chest x-ray as an ominous sign suggesting mediastinal abscess. PMID- 8223295 TI - Antrochoanal polyposis: a review of 33 cases. PMID- 8223296 TI - Symptom relief after endoscopic sinus surgery: an outcomes-based study. PMID- 8223297 TI - Symptom relief after endoscopic sinus surgery: an outcomes-based study. PMID- 8223298 TI - How the FDA has contributed to mistreatment of pediatric rhinosinusitis. PMID- 8223299 TI - Laboratory services in primary health care. PMID- 8223300 TI - Laboratory quality control, quality assurance and laboratory management. PMID- 8223301 TI - External quality control performance in clinical chemistry: experience in Kenya. AB - Analysis of eleven biochemical laboratory tests was done during an International External Quality Assessment Scheme (IEQAS) in which the clinical chemistry laboratory at Kenyatta National Hospital participated. Technicon SMA II continuous flow system was used in the biochemical analyses apart from glucose which was assayed manually by the glucose oxidase method. Using the standard deviation index (SDI), twenty six percent of the results were found to be outside the two standard deviation (2SD) limit. However, when variance index score (VIS) was used, 42% of the results were found to be outliers. Overall, our laboratory performed poorly compared to other laboratories in both the IEQAS and the United Kingdom External Quality Assurance Scheme (UKEQAS). This poor performance is attributed to the use of improper equipment which is not regularly maintained, lack of diagnostic reagents, lack of quality control (QC) materials and inadequate staff training in the field of quality control. PMID- 8223302 TI - Quality control in haematology and blood transfusion in sub-Saharan region of Africa. PMID- 8223303 TI - Autopsies: their need including their role in medical education and contribution to society. PMID- 8223304 TI - Forensic autopsies, their need including their social value. PMID- 8223305 TI - Clinical and laboratory approach for the diagnosis of galactosaemia in Africa. AB - A new method free from haemoglobin interference is described to measure erythrocyte galactose-l-phosphate uridyltransferase(GALT) activity levels. Since haemoglobin is removed by acid denaturation and precipitation with chloroform, only a simple photometer is required to assay GALT activity, implying that this method can be performed even when only basic laboratory facilities are available. Using this method, four infants have been found to suffer from galactosaemia due to GALT deficiency. Three children had very low, but still measurable GALT activities (< 1.0 mumol glucose-l-phosphate formed/gHb/hr) while one child had no detectable erythrocyte GALT activity. Compared to the other three children, this latter infant was seriously ill and required intensive care treatment before the diagnosis of galactosaemia was made. These results confirm the existence of different GALT variants in galactosaemia, in which the variant with zero activity has the most serious clinical sequelae if not appropriately treated. PMID- 8223306 TI - Pathology in sub-Saharan Africa--past, present and future. PMID- 8223307 TI - Role of fine-needle aspiration biopsy in the diagnosis of breast lumps at the Kenyatta National Hospital. AB - During 1990-1991, all patients seen at the surgical out-patient clinic at the Kenyatta National Hospital (KNH) with breast lumps had a fine-needle aspiration biopsy for diagnosis prior to open biopsy. Out of 105 that had corresponding surgical biopsies, 82 aspirates were considered satisfactory. Of these, the correct diagnosis was made in 59 (71%), another 12 (21%) were called benign but the correct histologic diagnosis was not made, 7 (8.5%) were correctly called negatives. The diagnostic accuracy of this method in differentiating benign from malignant was 95% in this study. PMID- 8223308 TI - Hepatitis B virus, hepatocellular carcinoma and liver cirrhosis in Kenya. AB - Hepatocellular carcinoma is the third most common malignancy in Kenyan males occurring with a peak incidence at 40 years of age. A worldwide correlation has been noted between the incidence of hepatocellular carcinoma and prevalence of hepatitis B virus. Liver biopsies with histological diagnosis of hepatocellular carcinoma (HCC), cirrhosis and the normals were reviewed by the authors. They were then stained for hepatitis B surface antigen (HBsAg) and hepatitis e core antigen (HBcAg). Only 2.5% of normal livers were positive for HBsAg compared with 33% of HCC and 25% of cirrhosis respectively. Hepatitis core antigen was not demonstrated in normal liver biopsies but it was present in 11.5% of HCC and 14% of cirrhosis. Background cirrhosis was noted in 52% of biopsies showing HCC. It is clear that a causal association exists between hepatitis B virus (HBV) and both liver cirrhosis and hepatocellular carcinoma. Higher antigen markers, up to 80% have been reported in South East Asia and India. This difference may be due to the type of biopsy examined (needle biopsy vs open biopsy) but the possibility that other factors such as aflatoxin and non A/non B hepatitis viruses play a more significant role in the causation of liver disease in Kenya than has previously been assumed should be explored. PMID- 8223309 TI - Degenerative leiomyopathy in African children: a review of current perspectives. AB - The syndrome of degenerative leiomyopathy causing intestinal obstruction clinically manifests in young indigenous African children as massive megacolon without aganglionosis. Eighteen children have been seen over a 16 year period from a localized geographic area. There were 10 males and 8 females having a mean age of 9.5 years at presentation and a mean duration of symptoms of 4.3 years. The youngest was 6 months old. All had progressive abdominal distension and infrequent stooling but 11 had intermittent diarrhoea and 9 had colicky abdominal pain. Gross gaseous distension of the large bowel with extension into small intestine occurred in 9 and this extended into stomach and oesophagus in 4. Biopsy of the dilated, thin walled bowel showed smooth muscle degeneration and necrosis with replacement by fibrous tissue. Neuronal cells of Auerbach's plexus tend to be displaced into the circular layer of smooth muscle with mild inflammatory changes. Some small arteries show medial fibrosis with subintimal fibroblastic proliferation. Acetylcholinesterase and immunohistochemical staining with neural and muscle markers are within normal limits. One child died while 14 have been maintained on prokinetic agents, low residue diets, laxatives and enemas. Nine children have required surgical intervention of whom 4 had volvulus and 3 adhesive bowel obstruction. Degenerative leiomyopathy is a distinctive entity with classical clinical and histological features. The aetiology is still obscure. PMID- 8223310 TI - Glucose 6-phosphate dehydrogenase deficiency in Africa--review. PMID- 8223311 TI - Pathology services in east, central and southern Africa. PMID- 8223312 TI - Persistent pulmonary hypertension in the newborn. PMID- 8223313 TI - The skinfold thickness in preterm infants. AB - The development of the subscapular, triceps and abdominal (suprailiac) skinfolds was studied in 120 healthy preterm infants with a birth weight below 2500 g at the age of 2 weeks, 1 month and 2 months. The skinfolds developed with the age of the infants, no gestational age related differences being noted. The triceps skinfold was thicker than the subscapular and abdominal skinfolds. The increase of skinfold thickness in identically-fed healthy appropriate-for-gestational-age preterm infants correlated with post-conceptional age. Small-for-gestational-age babies had thinner skinfold thicknesses up to a post-conceptional age of 44 weeks. PMID- 8223314 TI - Behavioural states and Doppler velocimetry of the renal artery in the near term human fetus. AB - Transition of fetal behavioural state 1F to 2F coincides with cardiovascular changes measured by Doppler velocimetry e.g. a decrease in pulsatility index (PI) in the internal carotid arteries and in the descending aorta, indicating redistribution of blood flow. Recently, we reported a considerable reduction in fetal urine production rate of 47% during fetal behavioural state 2F as compared to 1F. It was suggested that this reduction is caused by an increase in renal vascular resistance during 2F. Using Doppler ultrasound, flow velocity waveforms (FVW) of fetal renal arteries were recorded during behavioural states 1F and 2F. Fetal behavioural states 1F and 2F were assessed by recording fetal heart rate pattern, eye movements and body movements. The PI of the renal artery FVWs were calculated as an index of renal vascular resistance to blood flow. Fifteen healthy pregnant women between 36 and 40 weeks gestational age were studied and the relationship between fetal renal vascular resistance to blood flow and behavioural states was determined. We found that PI values in the renal arteries did not change relative to behavioural state 1F and 2F. These findings suggest that renal vascular resistance to blood flow is not appreciably different in 1F and 2F. This is in contrast with urine production rate which is almost reduced by half during 2F in the near term fetus. PMID- 8223315 TI - Longitudinal study of plasma ACTH and cortisol in very low birth weight infants in the first 8 weeks of life. AB - There are few published data on plasma ACTH and cortisol in very low birth weight (VLBW) infants beyond the first week of life. We therefore measured plasma ACTH and cortisol longitudinally in 25 infants (mean birth weight 1025 g, mean gestational age 28 weeks) at 1, 2, 4 and 8 postnatal weeks to document normative values for infants not receiving dexamethasone. We also examined the influence of clinical state and dexamethasone treatment on plasma ACTH and cortisol levels. Median plasma ACTH increased significantly with advancing postnatal age from 1 week to 8 weeks (21.0 vs. 40.0 ng/l; P = 0.01) but did not correlate with postconceptional age. Median plasma cortisol decreased significantly with advancing postnatal age from 1 week to 8 weeks (216 vs. 50 nmol/l; P = 0.001) and correlated inversely with postconceptional age (P = 0.004). At 8 weeks infants who were clinically well (n = 6) had lower plasma ACTH values compared with sick (n = 6) infants (median: 37.0 vs. 63.5 ng/l; P = 0.033). Plasma ACTH did not correlate with clinical state at 1, 2 and 4 weeks. At none of the postnatal ages studied was plasma cortisol influenced by the degree of sickness. Five infants received dexamethasone to assist weaning from mechanical ventilation. Their median plasma ACTH level, at 8 weeks, was significantly lower than that of the 12 infants who did not receive dexamethasone (11.0 vs. 40.0 ng/l; P = 0.0006). Plasma cortisol was not significantly influenced by dexamethasone treatment (P = 0.27). These data provide further information on the evolution of adrenocortical function in VLBW infants in the first months of life. PMID- 8223316 TI - Response of the premature fetus to stimulation by speech sounds. AB - The response of the premature fetus to speech stimuli was studied in 41 healthy pregnant patients at 26-34 weeks gestation. Speech stimuli consisted of repeated syllables ('ee' and 'ah') presented externally over the maternal abdomen at either 100, 105, or 110 decibels (dB). Sound stimuli were delivered during periods of both high and low fetal heart rate variability. During periods of low FHR variability, a decrease in fetal heart rate and an increase in the standard deviation of heart rate were found. During periods of high FHR variability, no significant change in either of these measures was observed. This is the first clear demonstration of heart rate responses to speech stimuli in the premature fetus. As is the case in the term fetus, this response is dependent on baseline heart rate variability which is the primary determinant of fetal state. The clinical usefulness of this finding may be limited by the magnitude of the response. PMID- 8223317 TI - Assessment of non-invasive techniques for measuring blood pressure in preterm infants of birthweight less than or equal to 750 grams. AB - Non-invasive techniques for measuring systolic blood pressure (BP) were evaluated in 10 very preterm infants, median gestational age 24 weeks, who were all of birthweight < or = 5750 g. Systolic BP measurements were attempted in all infants using a Doppler technique (Ultrasonic Doppler Flow Detector) and two oscillometers (Sentry and EME). Using each technique five separate measurements were attempted over a 10-min period. The systolic BP measurements for each device were then compared to results obtained from intra-arterial catheters which had an undamped waveform. BP measurements were possible on all 10 infants using the Doppler technique, but only in seven using the EME oscillometer and four using the Sentry oscillometer. The mean difference in measurements from those obtained using the arterial catheter were 0.27 mmHg using the Doppler technique and, in those infants in whom measurements were possible, 1.34 mmHg using the Sentry oscillometer and 1.34 mmHg using the Sentry oscillometer. We conclude that the Doppler technique provides the most useful and reliable non-invasive method of assessment of systolic BP in immature infants of birthweight < or = 750 g. PMID- 8223318 TI - [Trends and potential for human health resources research in Brazil]. PMID- 8223319 TI - Health advocacy post graduate education in Brazil: a response to new constitutional rights. PMID- 8223320 TI - [The administration of knowledge as a strategic orientation of PAHO/WHO]. PMID- 8223321 TI - [Human resources in health: current status]. PMID- 8223322 TI - [Implementation of quality assurance programs in the hospitals of Latin America and the Caribbean]. PMID- 8223323 TI - [Evaluation in municipal medical residencies: clinical pediatrics at the Hospital Pedro de Elizalde]. PMID- 8223324 TI - [Development of a scale of beliefs about social medicine: Dr. Pablo Acosta Ortiz Medical School]. PMID- 8223325 TI - [The PAHO Human Resources Development series]. PMID- 8223326 TI - [Effectiveness of a community program of teaching and care in oral health]. PMID- 8223327 TI - [Evaluation of basic knowledge for the management of infectious diseases: results of a survey]. PMID- 8223328 TI - [Preference for general medicine in a medical program: relation to change in the clinical environment]. PMID- 8223329 TI - [Application of a model of primary oral health care with a teaching/care-giving focus]. PMID- 8223330 TI - Pyridostigmine toxicity. Electrophysiological study. AB - A 54-year-old neurotic patient developed pyridostigmine-induced proximal weakness with daily 3600 mg of pyridostigmine. Detailed analysis of the RNS test showed three different types of responses which were reproduced in the in vitro nerve muscle preparation, confirming that three different responses represent the different degree of pyridostigmine toxicity. One of responses was a pattern, thought to be typical of anticholinesterase toxicity: repetitive discharge and the maximal decrement in the second response followed by an increment (dip phenomenon). With reduction of pyridostigmine, clinical and electrophysiological improvement followed. PMID- 8223331 TI - Alterations of motor evoked potentials by thalamotomy. AB - To evaluate the effect of stereotactic thalamotomy on the function of the corticospinal tract, we studied motor evoked potentials (MEPs) recorded by surface electromyography (EMG) in the left extensor carpi radialis (ECR) and flexor carpi radialis (FCR) with magnetic stimulation of the contralateral motor cortex in a 43-year-old patient with a severe postural and resting tremor of the left hand. The patient was diagnosed eight years previously with left hemiparkinsonism. The tremor was unresponsive to various medications. After thalamotomy the tremor had disappeared, confirmed by EMG studies. MEP latencies at rest were normal and did not change after thalamotomy. Volitional contraction of either ECR or FCR shortened the latency of the corresponding MEP before and after thalamotomy. However, before thalamotomy responses at rest were less well synchronized and followed by EMG silence with subsequent long duration tonic after discharges. Furthermore, during voluntary contraction the responses only slightly enhanced. After surgery MEPs at rest in both muscles were more synchronized and after-discharges had disappeared. Moreover, with volitional contraction of either ECR of FCR, the MEPs enhanced more dramatically. The silent periods (SPs) following the MEP during sustained voluntary contraction were longer after thalamotomy. The consistent MEP latencies suggest that the conduction of the pyramidal tract is unaffected by thalamotomy. The better synchronized responses, the alleviation of after-discharges and the longer SPs in this patient with hemiparkinsonism following thalamotomy suggest an improved sensorimotor integration, which may be the result of a reduced thalamic input onto suprasegmental levels. PMID- 8223332 TI - Combined ultrasonographic and neurographic examination: a new technique to evaluate phrenic nerve function. AB - In this report a new technique for simultaneous assessment of the electrical and mechanical properties of the diaphragm is presented. The phrenic nerve of 9 healthy persons was stimulated at the neck using magnetic stimulation. Electrical activity was recorded using chest surface electrodes. The mechanical function of the diaphragm was evaluated by performing combined B- and M-mode ultrasonographic examination. The data show that this method helps to distinguish the diaphragm muscle compound action potential from electrical activity derived from neighboring muscles activated by the brachial plexus. Furthermore, it provides a non-invasive tool which can easily be used in clinical practice to study the mechanical properties of the diaphragm. PMID- 8223333 TI - A modern representation of neuromuscular excitability in the form of intensity duration curve or line. AB - In 1939 Katz had mathematically defined the intensity-duration curve, and it has been used in this form to this day. In 1990, Ljubin and Knesaurek proposed the new mathematical and graphic form of the intensity-duration curve as a line. The new form, both graphic and mathematical, is considered to be more accurate and it is a new contribution to research into neuromuscular excitability. Besides, this method also makes possible a modern, numerical presentation of the clinical results and gives a new insight into observation of neuromuscular excitability, both in physiological conditions and pathology. Stephen's graphs, as well as of other authors, showing the intensity duration curves can also be transformed into lines, by translating them into the coordinate system with both logarithmic axes. PMID- 8223334 TI - Quantitation of tendon reflexes in normal volunteers. AB - Ankle (ATR), patellar (PTR) and triceps (TTR) T-reflex responses were recorded in 48 limbs of 24 normal volunteers by delivering tendon taps with a hand operated electronic hammer. Latency to the onset of the triggered response, peak to peak amplitude and duration of the wave were measured after several repetitions. In all individuals, ATR and PTR were observed, while in 27% of limbs, TTR was absent. Shortest latency and duration of the elicited response were chosen to calculate normal means. Side differences were also evaluated. As amplitude showed a marked interindividual and side to side variation, lowest recorded value was selected to represent the lower limit of normal. Age, height, and leg length showed a good correlation with minimum ATR latency (p < 0.001). PTR latency correlated significantly with age, height, (p < 0.001) and thigh length (p < 0.001). No significant correlation existed between age, height, arm length and TTR latency (p > 0.01). In conclusion, ATR and PTR are acceptable tests in clinical practice, whereas TTR is not reliable, as it is technically difficult to elicit in some normal individuals. PMID- 8223335 TI - Electrophysiological study on diabetic neuropathy treated with Cronassial. PMID- 8223336 TI - A prospective one-year follow-up study with somatosensory potentials evoked by stimulation of the median nerve in patients with cerebral infarct. AB - Somatosensory potentials evoked by stimulation of the median nerve (median nerve SEPs) were studied in a prospective and sequential series of 40 patients with first supratentorial and nonhaemorrhagic cerebral infarct. In 35 patients the SEPs were recorded three times during the first year after the stroke. The location of the infarcted zone was reflected in the number of detected abnormalities: most patients with infarct changes extending to the gray matter of the Rolandic cortex showed abnormalities in the median nerve SEPs, and all patients with involvement of both precentral and postcentral cortical gray matter had abnormal median nerve SEPs. In the entire patient group when both latency and amplitude abnormalities were included about half (48%) of the patients had abnormal median nerve SEPs a week after the stroke, 39% 2-3 months after the stroke and 29% about one year after the stroke. These changes were not significant. When separately surveying the changes in the numbers of latency and amplitude abnormalities the difference between the first and the third examinations was nearly significant only in the number of latency abnormalities. Furthermore, in the absolute latency and amplitude values, no significant changes could be seen within the first two-three months after the infarct; within the whole one-year follow-up period a nearly significant change was noted between the second and the third examination in only one parameter (P22 peak latency). Thus, the abnormalities in the median nerve SEPs, especially the amplitude abnormalities, were relatively permanent during the one-year period after cerebral infarct. PMID- 8223337 TI - The value of electrophysiological examination of the flexor carpi ulnaris muscle in the diagnosis of cubital tunnel syndrome. AB - In the electrodiagnosis of cubital tunnel syndrome, the abductor digiti minimi (ADM) muscle is frequently used to evoke compound muscle action potentials (CMAP) in order to measure the motor nerve conduction velocity (MNCV) in the ulnar nerve. Slowing of the MNCV across the elbow is a major criteria in the electrodiagnosis of cubital tunnel syndrome. However, in advanced cases, slowing of the MNCV is also seen in the forearm segment, thus, localization of the nerve palsy is sometimes difficult. We evoked CMAP from the flexor carpi ulnaris (FCU) muscle to assess the value of electrophysiological examination of this muscle in the diagnosis of cubital tunnel syndrome. CMAP from the FCU muscle could be evoked in many cases when CMAP from the ADM muscle could not. Also, the motor terminal latency (MTL) of the FCU was prolonged in proportion to the slowing of the MNCV across the elbow. Therefore, electrophysiological examination of the FCU muscle is useful in the diagnosis of cubital tunnel syndrome, especially when intrinsic muscle atrophy is severe and the CMAP from these muscles cannot be evoked. PMID- 8223338 TI - Features of motor control in patients with proximal childhood spinal muscle atrophy (pilot study). AB - The differences in the motor performance during different tasks between 19 subjects suffering from SMA and 10 healthy controls were observed. The simultaneous EMG activity of twelve lower limbs and lower trunk muscles was recorded with surface electrodes. EMG data were automatically reduced and compared with data evaluated from performed by physiotherapist manual testing of muscle strength. Results showed characteristic differences between healthy and spinal muscular atrophy (SMA) subjects: 1. SMA patients display generally more activity occurring in numerous muscle groups and more spinal levels are activated. 2. SMA patients reveal a disturbed functional relation between the posterior and anterior compartments of muscles. 3. EMG activity in SMA patients is spreading out also to the contralateral muscle groups even during slight, unilateral singlejoint movements. Oligosegmental, plurisegmental and brain sources are probably responsible for mentioned phenomena. The reciprocal influences between reduced number of motoneurons (in SMA) and function of central movement generators results in different mode of movement execution in SMA patients. PMID- 8223339 TI - The use and misuse of thyroid hormone. PMID- 8223340 TI - Molecular mechanisms mediating proliferation/differentiation interrelationships during progressive development of the osteoblast phenotype. PMID- 8223341 TI - Steroid hormone receptors: interaction with deoxyribonucleic acid and transcription factors. AB - Gene regulation by steroid hormones is mediated by binding of the hormone ligand to the corresponding receptor that triggers a complex set of interactions of the hormone receptors with each other, with DNA in chromatin, and with a variety of other proteins. In this review we attempt to summarize what is known about these interactions using as the main example the regulation of mouse mammary tumor virus transcription by glucocorticoids and progestins. We describe in some detail the interaction of monomers and homodimers of the steroid receptors with their recognition sequences, and the molecular mechanism used to discriminate between the responsive elements for glucocorticoids/progestins and estrogens. We then review the interactions between homologous and heterologous hormone receptors on complex hormone regulatory regions, before devoting some attention to the synergistic and inhibitory interactions of hormone receptors with other transcription factors. Finally we briefly summarize some of the possible mechanisms that modulate the molecular interactions of hormone receptors. In addition to ligand binding, these include receptor phosphorylation, changes in DNA topology, and the organization of DNA in nucleosomes. From this overview we draw the tentative conclusion that the specificity of the hormonal response in different cells results from a combination of developmental restrictions both in the accessibility of genomic sequence and in the repertoire of regulatory proteins present in each particular cell. In addition, the array of regulatory sequences in DNA and chromatin determines the precise nature of macromolecular interactions of the receptors that are modulated by their degree of phosphorylation. PMID- 8223342 TI - Serotonin, a neurotransmitter involved in the regulation of luteinizing hormone release. AB - The involvement of serotonin (5-HT) in the regulation of LH secretion is discussed on the basis of experimental and physiological models. The role of 5-HT on low amplitude pulsatile LH release in male rats is not yet clear, in spite of the fact that recent results suggest a weak permissive role. In ovariectomized rats, 5-HT expresses a negative influence on the increased rate of LH release, which is converted into a stimulatory effect by pretreatment of the animals with E2. Moreover, in castrated female rats there is a morning/afternoon oscillatory pattern in 5-HT metabolism of brain areas associated with the control of LH secretion including the hypothalamus. In this area, the fluctuation is modulated by E2 in such a way that the peak of 5-HT neural activity occurs simultaneously with the induced afternoon discharge of LH. On the other hand, P enhances the amplitude of the oscillation of hypothalamic 5-HT metabolism concomitantly with a potentiation of the induced LH surge. This facilitatory role of 5-HT upon phasic LH discharge is also evident in intact female rats. The preovulatory surge of LH is accompanied by an increased hypothalamic and, more precisely, ME 5-HT turnover. Furthermore, 5-HT stimulates in vitro LHRH release from the ME. Serotonergic nuclei located in the brain stem seem to mediate this effect. PMID- 8223343 TI - Citation for the Fred Conrad Koch Award of the Endocrine Society to Jean D. Wilson. PMID- 8223344 TI - Citation for the Edwin B. Astwood Lecture Award of the Endocrine Society to Ronald M. Evans. PMID- 8223345 TI - Citation for the Rhone-Poulenc Rorer Clinical Investigator Lecture Award of the Endocrine Society to William L. McGuire. PMID- 8223346 TI - Citation for the Sidney H. Ingbar Distinguished Service Award of the Endocrine Society to Gabriel Bialy. PMID- 8223347 TI - Citation for the Gerald D. Aurbach Award of the Endocrine Society to R. Daniel Camerini-Otero. PMID- 8223348 TI - Citation for the Robert H. Williams Distinguished Leadership Award of the Endocrine Society to Willard P. VanderLaan. PMID- 8223349 TI - Citation for the Ernst Oppenheimer Award of the Endocrine Society to Margaret A. Shupnik. PMID- 8223350 TI - Citation for the Richard E. Weitzman Memorial Award to Daniel J. Drucker. PMID- 8223351 TI - Haemodynamic effects and comparison of enoximone, dobutamine and dopamine following mitral valve surgery. AB - Mitral valve surgery may be complicated by a post-operative low output state requiring inotropic support, and a wide variety of factors may influence the choice of agents used to treat this condition. The authors have examined and compared the haemodynamic effects of the highly specific phosphodiesterase inhibitor enoximone, and the adrenergic agents dobutamine and dopamine in patients undergoing mitral valve surgery. Enoximone, 0.5 mg kg-1 bolus, followed by a continuous infusion of 5 micrograms kg-1 min-1, was compared against dobutamine, 7 micrograms kg-1 min-1, and dopamine, 5 micrograms kg-1 min-1, with the protocol allowing for an increase in the infusion rate by a factor of two if clinical and haemodynamic measurements indicated. All 25 patients receiving enoximone were successfully weaned from cardiopulmonary bypass at the first attempt, with significant increases in cardiac index and stroke index, combined with little or no change in heart rate or pulmonary artery pressures and a highly significant reduction in systemic vascular resistance, and a reduction in mean arterial pressure. Three of the 25 patients receiving dobutamine were withdrawn from the study because of inadequate haemodynamic response, while the remaining 22 patients demonstrated significant increases in heart rate, cardiac index and stroke index, with a reduction in systemic vascular resistance. Nine of the 25 patients receiving dopamine failed to respond adequately, while the remaining 16 demonstrated an increase in heart rate and cardiac index but with little change in stroke index and a modest reduction in systemic vascular resistance. Enoximone has been shown to be a highly effective first-line inotrope in patients following mitral valve surgery with significant advantages over dobutamine and dopamine. PMID- 8223352 TI - Experience with phosphodiesterase inhibitors in paediatric cardiac surgery. AB - In a controlled study of newborns with refractory low-output states (n = 16), a bolus dose of enoximone, 1 mg kg-1min-1, produced a dramatic haemodynamic improvement in 12 of 16 patients ('responders'). Of these 12, 9 survived. All 'non-responders' died subsequently. Enoximone resulted in reduced filling pressures (P < 0.005) and improved cardiac index (before enoximone, 0.96 litres min-1 m-2; after enoximone, 3.05 litres min-1 m-2; P < 0.001). To assess the routine use of enoximone in paediatric surgical patients, enoximone was administered post-operatively in 84 children in addition to standard therapy during 1991-92. Clinical improvement was noted in 58 of 84 patients (69%) and no significant side-effects were observed. It is concluded that enoximone provides a safe and effective pharmacological approach for the management of cardiac low output states in children. PMID- 8223353 TI - Improving morbidity and mortality after cardiac surgery. AB - Over the past three decades there has been an enormous increase in the number of cardiac operations, particularly on patients with coronary artery disease. Technical advances in cardiac surgery, anaesthesia, monitoring and intensive care would have been expected to reduce morbidity and mortality, but the incidence of complications has, in fact, increased. The increasing use of coronary angiography and the development of interventional cardiology have caused a change in the type of patients scheduled for surgery--they have become progressively older and sicker than in the 1970s. Consequently, the management of patients has to be revised regularly. Careful selection of patients, and the defining of specific strategies for anaesthetic and surgical management are required to improve the outlook for post-surgical complications. PMID- 8223354 TI - What is the place of the phosphodiesterase inhibitors? AB - Pharmacological support is often required during and after weaning from cardiopulmonary bypass. Most standard therapy acts on alpha-, beta- or dopaminergic-receptors, but down-regulation of beta-receptors has led to the development of agents which act independently from the beta-receptor. The phosphodiesterase (PDE) III inhibitor enoximone, was evaluated in cardiac surgery. Application of enoximone prior to weaning from CPB improved overall cardiac function, reduced the need for catecholaminergic inotropic support, and provided increased organ perfusion even 2 h after the operation. 'Pre-treatment' with enoximone may be beneficial, due to its ability to combine inotropic support with systemic and pulmonary vasodilatation, and may be considered as a first-line approach instead of catecholamines. PMID- 8223355 TI - Cost-effectiveness in patients awaiting transplantation receiving intravenous inotropic support. AB - Cost-effectiveness, assessed in terms of cost/survivor and cost/additional day of survival, has been evaluated in two groups of patients awaiting cardiac transplantation: in six treated by mechanical support (mechanical bridge) and in 31 treated successfully by the addition of intravenous enoximone to previous existing inotropic support (pharmacological bridge). The mean cost per patient was $45,843, ranging from $38,326 in patients receiving pharmacological support to $84,683 in patients receiving mechanical support. The cost per patient transplanted after pharmacological support was $50,745. The mean cost per survivor at 1 year was $210,000 for all patients, ranging from $192,455 with pharmacological support to $254,000 with mechanical support. At 1 month, the cost per additional day of survival was higher (by 228%) in the mechanical support group than in the pharmacological support group. PMID- 8223356 TI - Whether obesity should be treated. AB - Antidieting sentiment is strengthened in part by the assumption that all diets fail, which in turn is based on data from university-based treatment programs. A small percentage of overweight people attempt to lose weight in these programs, and they differ in important ways from overweight people in general. They are heavier, more likely to be binge eaters, and have higher levels of psychopathology--all negative prognostic factors for treatment. Generalizing from the treatment of this group to all programs is highly questionable. Hence, little is known about the effectiveness of approaches used by most people. There is a pressing need for research on this topic, and a need for treatments that address the specific needs of those who seek clinical treatments. PMID- 8223357 TI - Obese people who seek treatment have different characteristics than those who do not seek treatment. AB - A group of obese people who had not sought treatment, an obese group who had sought treatment in a professional, hospital-based program, and normal-weight controls (N = 547) were compared in regard to level of psychopathology, binge eating, and negative emotional eating. Because the groups differed significantly on several demographic variables, 3 demographically matched groups were created and compared (n = 177, 59 per group). In the matched subgroups, obese people who had sought treatment reported greater psychopathology and more binge eating than did those who had not sought treatment or did normal-weight controls. Both obese groups (including those who had not sought treatment) endorsed more symptoms of distress, negative emotional eating, overeating, difficulty resisting temptation, and less exercise than did normal-weight controls. PMID- 8223358 TI - Condom use: a self-efficacy model. AB - A survey of heterosexually active college students gathered information about condom use, self-efficacy (SE), outcome expectancies, sexual attitudes, peer group influences, acquired immunodeficiency syndrome (AIDS) knowledge, and perceived vulnerability to AIDS. On the basis of A. Bandura's (1986) social cognitive theory, a structural model with SE as the central mediator was formulated and evaluated with LISREL. This model explained 46% of the variance in condom use from judgments of SE and effects attributable to peers and 53% of the variance in SE from outcome expectancies and peer group influences. Sexual attitudes, AIDS knowledge, and perceived vulnerability did not predict condom use. Most students were well-informed about human immunodeficiency virus (HIV) transmission but reported not feeling at risk, even though many engaged in risky sexual behavior. PMID- 8223359 TI - Race and gender comparisons: I. Hemodynamic responses to a series of stressors. AB - A sample of 155 adults, age 18-49 years, including nearly equal subgroups of Black and White men and women, underwent evaluation of cardiovascular reactivity during 5 behavioral stressors. Among the men, overall blood pressure increases to tasks did not differ, but Blacks showed generally higher total peripheral resistance, whereas Whites showed greater heart rate and cardiac output increases. Among the women, the same racial-group differences were evident during certain tasks, but not during others. Men showed greater overall systolic blood pressure increases than did women, and they also showed less recovery toward baseline levels in systolic and diastolic pressure and stroke volume 5 min after the stressors. Other gender differences were task specific. The possible contributions of structural changes in the myocardium and vasculature, of altered sympathetic receptor distribution, and of task-specific behavioral factors influencing task involvement are discussed. PMID- 8223360 TI - Race and gender comparisons: II. Predictions of work blood pressure from laboratory baseline and cardiovascular reactivity measures. AB - In 148 Black and White men and women, laboratory measures of blood pressure (BP), heart rate, stroke volume (SV), cardiac output (CO) and total peripheral resistance (TPR) during baseline and 5 stressors were examined in relationship to ambulatory systolic (SBP) and diastolic (DBP) blood pressures at work. Baseline BP strongly predicted mean work levels in all groups. For White men and Black women, higher SV and CO responses to the active speech and averaged across all tasks predicted higher work SBP individually and also when added to a model based on baseline SBP, age, and diary information. For White women, higher SBP increases to the passive speech similarly predicted mean work SBP. For Black men, higher TPR response to the cold pressor test correlated with higher work SBP but did not improve a predictor model involving baseline SBP and age. Reactivity measures did not consistently contribute to prediction of work DBP. PMID- 8223361 TI - Sex differences in social support, self-deception, hostility, and ambulatory cardiovascular activity. AB - The quality of the social network, ambulatory cardiovascular indices, and other health risks were assessed in 129 students. Ss carried an ambulatory monitor for a working day and completed a social support scale, an index of daily stress, and measures of anger expression, hostility, depression, and self-deception. Alcohol intake and skinfold thickness were assessed. Results indicated that quality of social support was related in an inverse manner to ambulatory systolic pressure in women only. Stepwise multiple regression confirmed that social support was an independent predictor of daily systolic pressure for women and was not confounded with subjective stress. For men, tendencies to self-deception and high hostility were independently related to elevated blood pressure and heart rate. PMID- 8223362 TI - Emotional and sensory focus as mediators of dental pain among patients differing in desired and felt dental control. AB - Giving patients instructions to focus on sensory (vs. emotional) stimuli during a root canal procedure significantly reduced self-reported pain, but only among patients who were classified as having strong desire for control and low felt control in dental situations. Among patients with low felt control and low desire for control, sensory-focus instructions produced greater pain reports than did emotion-focus instructions. Finally, high desire-low felt patients reported higher levels of expected pain before treatment than did other patient subgroups. These data suggested limiting conditions for H. Leventhal's (1982) theory of emotion and supported the idea that desire for control might moderate the effects of perceived control. PMID- 8223363 TI - Correlates of physical activity at home in Mexican-American and Anglo-American preschool children. AB - Twenty-two potential correlates of children's physical activity were examined. Two hundred and one Mexican-American and 146 Anglo-American families with 4-year old children were studied. Children's physical activity was directly observed in the evening at home on 4 visits for 1 hr each time. Anglo-American children and male children were found to be more active. Demographic variables explained 11% of the variance in children's physical activity. After adjusting for demographics, 3 children's variables and 6 social-family variables did not account for significantly more variance. Five environmental variables accounted for 11% additional variance. Variables observed concurrently with physical activity, such as time spent outdoors and prompts to be active, were highly associated with children's physical activity. PMID- 8223364 TI - Standardized, individualized, interactive, and personalized self-help programs for smoking cessation. AB - Smokers (N = 756) were randomly assigned by stage of change to (a) standardized self-help manuals (ALA+ condition), (b) individualized manuals matched to stage (TTT condition), (c) interactive expert-system computer reports plus individualized manuals (ITT condition), or (d) a personalized condition with 4 counselor calls, stage manuals, and computer reports (PITT condition). Over 18 months, the ITT group's results more than doubled those of the ALA+ group on abstinence measures. The ALA+ and TTT conditions were equivalent over 12 months, but at 18 months the TTT condition was more effective. The ITT condition was the best or comparable with the best treatment at all follow-ups for smokers at all stages of change. Results suggest that an effective expert system has been developed, and discussion focuses on delivering this system to entire populations of smokers. PMID- 8223365 TI - Construct validation of optimism and pessimism in older men: findings from the normative aging study. AB - Validation of Scheier and Carver's (1985) Life Orientation Test (LOT) has identified associations between bipolar optimism and several external constructs. However, optimism and pessimism may be not bipolar, but rather separate constructs. Furthermore, these constructs may be indistinguishable from personality traits, such as neuroticism and extraversion. This study examined the associations of separate optimism and pessimism measures with self-reports of hassles, psychological symptoms, and illness severity, controlling for personality. Ss were 1,192 men from the Normative Aging Study. Findings suggest that optimism and pessimism are separate and that their relations to external criteria remain, although attenuated, when neuroticism and extraversion are controlled. PMID- 8223366 TI - Diet, alcohol, and physical activity as a function of smoking status in middle aged women. AB - Diet, alcohol intake, and leisure-time physical activity were compared cross sectionally in middle-aged female smokers, ex-smokers, and never smokers and were reassessed approximately 3 years later. At initial contact, there were no group differences in total caloric intake and very few differences in nutrient intake, but alcohol intake of current smokers and ex-smokers was at least 50% greater than that of never smokers, and current smokers reported less physical activity than did ex-smokers and never smokers. Prospective findings were similar: Compared with continuing smokers, ex-smokers did not change their dietary and alcohol intake but significantly increased physical activity. Thus, some adverse behaviors associated with smoking (e.g., alcohol intake) may be due to self selection to history of smoking, whereas other behaviors (e.g., reduced physical activity) may be more directly related to smoking itself. PMID- 8223367 TI - Behavioral and cardiovascular responses to interpersonal challenges among male offspring of essential hypertensives. AB - Behavioral, cardiovascular, and self-report of cognitive and affective responses to 2 interpersonal challenges were examined among 20 men with a positive (FH+) and 20 with a negative (FH-) family history of hypertension. Heart rate (HR) and blood pressure (BP) were measured throughout the laboratory session; Ss were requested to self-report positive and negative cognitions, state anger, and state anxiety that occurred during interactions with a male and female confederate. Behavioral responses to interpersonal tasks were videotaped, coded, and categorized into 4 major groupings (positive verbal, positive nonverbal, negative verbal, and negative nonverbal). FH+ individuals exhibited significantly higher resting HR and systolic BP (SBP) reactivity to both interactions than FH- counterparts. Analyses of behavioral responses for both interactions revealed significantly more negative verbal and nonverbal behavior and less positive nonverbal behavior among FH+ as compared with FH-Ss. PMID- 8223368 TI - Sex differences in sensitivity of the hypothalamic-pituitary-adrenal axis. AB - Two studies examined sex differences in responsiveness of the hypothalamic pituitary-adrenal cortical axis, a major component of the stress response. The first measured pituitary-adrenal responses to ovine corticotropin-releasing hormone (oCRH) in 24 health men and 19 healthy women. Plasma adrenocorticotropin hormone (ACTH) response to oCRH were significantly greater among women than among men. In contrast, cortisol concentrations were similar in both groups, though elevations were more prolonged in women. Differences in corticotropin-releasing activity between men and women may help account for these findings; such differences in central components of the stress response might play a role in the known epidemiological differences in diseases of stress system dysregulation between men and women. PMID- 8223369 TI - The significance of cardiac auscultatory findings in horses: insight into the age old dilemma. PMID- 8223370 TI - Viruses, tumours and the MHC. PMID- 8223371 TI - Report of the first international workshop on equine sarcoid. PMID- 8223372 TI - A survey of cardiac auscultatory findings in horses. AB - Auscultation was used to examine 545 horses for cardiac murmurs and some arrhythmias. The most significant finding was the high prevalence of right-sided holosystolic or pansystolic plateau-type murmurs compatible with a diagnosis of tricuspid regurgitation in National Hunt racing Thoroughbreds of all ages (16.4%). This condition was found in 4.7% of flat-racing Thoroughbreds, 3.7% of non-racing Thoroughbreds and Thoroughbred crosses, and in none of the ponies examined. High prevalences of left-sided early systolic murmurs (53%), left-sided early diastolic murmurs (23%), right-sided early diastolic murmurs (22%) and second degree atrioventricular block (23%) were found in racehorses of both types. PMID- 8223373 TI - Measurement of 24-h gastric pH using an indwelling pH electrode in horses unfed, fed and treated with ranitidine. AB - A glass combined pH-reference electrode was placed in the stomachs of 5 adult horses and pH was recorded every 6 mins for 24 h while (1) feed and all bedding materials were withheld for 24 h (unfed), (2) horses had free access to Timothy grass hay for 24 h (fed), and (3) horses had free access to Timothy grass hay and were treated with ranitidine, 6.6 mg/kg body weight, orally, every 8 h for 48 h (fed + ranitidine). There was a significant (P = 0.007) difference in median 24-h gastric pH amongst the 3 protocols, the value being 3.1 in fed horses and 1.55 in unfed horses (P = 0.05) and 4.6 in fed + ranitidine horses (P = 0.05 compared with fed horses). The percentage of pH readings < 2.0 was significantly greater (P = 0.05) in unfed (76%) than in fed (30%) horses, and in fed horses than in fed + ranitidine (9%) horses. The percentage of readings > 2.0, 3.0, 4.0, 5.0 and 6.0 were each significantly (P = 0.05) greater in fed + ranitidine horses than in fed horses, and in fed than in unfed horses. There was no difference (P = 0.13) between median gastric pH values from different times of day in unfed horses (00:00 to 06:00 h, 1.5; 06:00 to 12:00 h, 1.6; 12:00 to 18:00 h, 1.7; and 18:00 to 24:00 h, 1.5). We conclude that gastric acidity is greatest when horses do not have access to feed material and that ranitidine effectively suppresses gastric acidity in horses under conditions of free access to hay. PMID- 8223374 TI - Forelimb lameness associated with radiographic abnormalities of the cervical vertebrae. AB - Eight horses had forelimb lameness which was considered unrelated to primary forelimb pain, but was associated with radiographic abnormalities of one or more cervical vertebrae. There was no evidence of ataxia or weakness. The degree and character of the forelimb lameness varied between horses. In 4 horses, selective local analgesic techniques were used to rule out lameness associated with pain in the forelimb. In the other 4 horses, radiographic examination of the cervical region was performed on the basis of forelimb lameness seen in conjunction with neck stiffness and/or neck pain. Three horses had marked modelling of the synovial articular facet joints in the caudal cervical region; a 4th horse had modelling and a fracture involving the synovial articulation between the 4th and 5th cervical vertebrae. One horse had abnormalities of the intercentral articulation between the 7th cervical and 1st thoracic vertebrae and a discrete mineralised opacity dorsal to it. Two horses had large lucent zones in a vertebral body. One horse had a fracture of the body of the 7th cervical vertebra. Five horses were humanely destroyed, 2 returned to their previous level of activity after a prolonged period of rest and 1 was still being rested at the time of writing. PMID- 8223375 TI - Retrospective analysis of the results of 151 exploratory laparotomies in horses with gastrointestinal disease. AB - Of 149 horses that underwent 151 exploratory laparotomies for gastrointestinal disorders from September 1987 to May 1991, 107 (72%) were discharged from the hospital: 100 (66%) survived for > 7 months, 94 of which returned to their intended use. Survival rate (64/80) for horses with caecum/large colon obstruction was significantly (P = 0.003) higher than for horses with small intestinal obstruction (33/64). Prolonged surgery was associated with significantly (P < 0.001) lower survival rates than short surgical time. In the large intestine, survival rate (15/29) for strangulated obstructions was significantly (P < 0.001) lower than for simple obstructions (52/58). Generalised septic peritonitis (9 horses) and bowel obstruction associated with adhesions (8 horses) were the most frequent fatal post-operative complications. The rate (6/44) of post-operative adhesions after small intestinal obstruction was significantly (P = 0.006) higher than that (2/68) following large intestinal obstruction. The rate (8/55) of post-operative adhesion formation in horses that required enterotomy/enterectomy was significantly (P = 0.003) higher than that (0/57) in horses that did not require gut wall incisions. Incisional suppuration developed in 42 horses and occurred with a significantly (P = 0.028) higher rate (32/72) after caecum/large colon lesions than after obstruction at other sites, (10/42) but was not associated with known contamination at the time of surgery (P = 0.806). PMID- 8223376 TI - Diagnosis and prediction of cervical vertebral malformation in thoroughbred foals based on semi-quantitative radiographic indicators. AB - Over a period of 5 years on a central Kentucky Thoroughbred stud farm 132 foals were evaluated for the presence of cervical vertebral malformation (CVM). Lateral cervical vertebral radiographs were obtained on 70 standing foals. These radiographs were evaluated semi-quantitatively and scored numerically for the presence of stenosis of the vertebral canal, enlarged physeal growth plates, caudal extension of the dorsal border of the orifice of the vertebral canal, angular fixation, delayed ossification of bone and degenerative joint disease. From these scores a total CVM score was given for each set of radiographs. All foals were repeatedly examined for clinical signs of neurological disease. Foals with no clinical signs of CVM had maximal total CVM scores (mean +/- sd) of 5.7 +/- 2.8 and foals affected with CVM had maximal total CVM scores of 17.0 +/- 2.8. Determination of the total CVM score was non-invasive, inexpensive and a very accurate procedure for predicting and diagnosing the presence of CVM in Thoroughbred foals up to 1 year of age. PMID- 8223377 TI - Correlation of racing performance with blood lactate and heart rate after exercise in thoroughbred horses. AB - At 2 and 5 mins after an 800-m gallop, venous blood was collected from 26 Thoroughbred racehorses for measurement of blood lactate concentration, packed cell volume (PCV) and haemoglobin concentration. In addition, 14 racehorses were given a strenuous submaximal treadmill exercise test. Heart rates during and after exercise at 10 m/sec on a treadmill inclined at 5 degrees were recorded. Blood samples at 2 and 5 mins after exercise were used to measure PCV, blood and plasma lactate and ammonia concentrations. Results of each exercise test were compared with the retrospective performance of horses in races, using Timeform ratings. The results of the field tests were also compared with the performance of each horse in a race 2 days later. There were no significant correlations between any of the measurements taken after the field test and either subsequent race performance or Timeform rating. Heart rate 4 mins after treadmill exercise was significantly correlated with Timeform rating (r = -0.565, P < 0.05). Blood and plasma lactate concentrations 2 and 5 mins after treadmill exercise were all significantly correlated with Timeform. The highest correlations were with blood lactate concentrations 2 and 5 mins after exercise (r = -0.68, P < 0.01). There were no significant correlations between Timeform and heart rate during exercise at 10 m/sec, heart rates at 1, 3 and 5 mins after exercise, PCV and plasma ammonia at 2 and 5 mins, or the differences between lactate concentration in plasma or blood at 2 and 5 mins after exercise.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223378 TI - Immunocytochemical and lectin histochemical study of neuronal lesions in autonomic ganglia of horses with grass sickness. AB - Equine grass sickness (EGS) is a primary dysautonomia characterised pathologically by lesions in autonomic ganglia, enteric plexi and specific nuclei in the CNS. Immunocytochemistry and lectin histochemistry of the autonomic ganglia were used to determine whether abnormalities can be detected in specific proteins or cellular organelles. EGS ganglia contained a mixture of morphologically normal and abnormal neurons, the former appearing identical to cells from control animals. Affected cells showed marked disturbances in neurofilament (NF) proteins and beta-tubulin, major components of the cytoskeleton; in most neurons immunoreactivity was reduced or absent while the distribution was altered in the remainder. Staining for neuron-specific enolase, a pan-neuronal marker, was severely reduced or absent, as was reactivity for the catecholaminergic enzyme tyrosine hydroxylase. However, affected neurons showed a marked increase in dopamine-beta-hydroxylase (D beta H), another enzyme associated with noradrenaline synthesis. Wheat germ agglutinin and Griffonia simplicifolia B4 lectin histochemistry was used to label membranes of the Golgi apparatus, which stained as discrete curvilinear perinuclear profiles. All affected neurons showed abnormalities with either complete loss of reaction or amorphous centrally located lectin staining. The results indicate perturbation in a wide variety of cytoplasmic and cytoskeletal proteins. In the majority of instances there is a decrease in stainable protein; the increase in D beta H may indicate a failure to be transported down the axon with resultant accumulation in the perikaryon. Loss of a recognisable Golgi structure appears to be an early event in the neuropathology of EGS. PMID- 8223379 TI - Radiographic evaluation of ossification of the collateral cartilages of the third phalanx in Finnhorses. AB - Of 462 Finnhorses (age 1-20 years) evaluated and graded (scale 0-5) radiographically for ossification of the collateral cartilages of the 3rd phalanx of the front feet, 22.9% were totally free from sidebones (Grade 0), and 49.1% had minimal or mild ossification (Grade 1 or 2) at the base of 1 or more of the cartilages. Moderate ossification (Grade 3) was present in 10.2% of the horses, whereas ossification was advanced (Grade 4) or extensive (Grade 5) in 17.8%. In most horses > 1 cartilage was affected and 7.6% of the horses had separate centres of ossification in 1 or more of the cartilages. Large sidebones and separate centres of ossification were more common in females than in males (P < 0.001). The incidence of large sidebones was lower in young females (1-3 years) than in mares 4-6 years of age (P < 0.05), but did not increase significantly with age when young horses, 4-6 years of age, were compared with older ones. Grade 4 and 5 sidebones as well as separate centres of ossification were more common in the lateral than in the medial cartilage (P < 0.001), but no difference was observed in this respect between the front feet. PMID- 8223380 TI - Hypochoeris-associated stringhalt in North America. PMID- 8223381 TI - Failure of intra-articular anaesthesia of the antebrachiocarpal joint to abolish lameness associated with chip fracture of the distal radius. PMID- 8223382 TI - Immune-mediated haemolytic disease after penicillin therapy in a horse. PMID- 8223383 TI - Aortic origin of the right pulmonary artery and patent ductus arteriosus in a pony foal with pulmonary hypertension and right-sided heart failure. PMID- 8223384 TI - Malignant histiocytosis in an Arabian filly. PMID- 8223385 TI - A prospective study of septicaemia in colostrum-deprived foals. Julie A. Robinson, G. K. Allen, Eleanor M. Green, W. H. Fales, W. E. Loch and Christina G. Wilkerson. Equine Vet. J. (1993) 25 (3), 214-219. PMID- 8223386 TI - [Legionella infection]. PMID- 8223387 TI - [Wiedemann-Beckwith syndrome]. PMID- 8223388 TI - The combination of the analytical capacity of electrophoresis with the sensitivity and specificity of immunoassays makes the hybrid technology of protein blotting one of the most valuable. PMID- 8223389 TI - Protein blotting: the 2nd decade. Paper symposium. PMID- 8223390 TI - Structural characterization of blotting membranes and the influence of membrane parameters for electroblotting and subsequent amino acid sequence analysis of proteins. AB - Various blotting membranes were evaluated and correlated with the efficiency of electroblotting and the performance in the sequencing process. Structural parameters including specific surface area, pore size distribution, pore volumes, and permeabilities of different solvents lead to discrimination of the membranes relative to their accessible surfaces and membrane densities. Protein binding capacities as well as protein recoveries in electroblotting correlate with the specific surface areas. Almost quantitative retention of proteins during electroblotting from gels was obtained for membranes with a high specific surface area and narrow pores (Trans-Blot, Immobilon PSQ, Fluorotrans), whereas membranes with a relatively low specific surface area (Immobilon P, Glassybond) showed reduced recoveries of between 10-20% for the tested proteins. Initial yields and repetitive yields were compared for radioiodinated standard proteins that have been either electroblotted or loaded by direct adsorption. The results showed that the different permeabilities for solutions of the Edman chemistry have a major influence on initial yields. The glass fiber-based membranes with an extremely low flow restriction produce consistently high initial yields independent of the application mode of the protein (spotted or electroblotted) or the application of the membranes into the cartridge (discs or small pieces). In contrast, the polymeric membranes showed decreasing initial yields with increasing membrane density for spotted and electroblotted proteins. Yields varied considerably when the membranes were applied as discs into the cartridge. This effect could be minimized by cutting the membranes into pieces as small as possible, as demonstrated for electroblotted proteins. PMID- 8223391 TI - Microsequence analysis of the N-terminally blocked proteins immobilized on polyvinylidene difluoride membrane by western blotting. AB - A technique has been developed for efficient deblocking and subsequent microsequencing of N-terminally blocked proteins immobilized on a polyvinylidene difluoride (PVDF) membrane at the picomole levels. In this technique, proteins were first separated by polyacrylamide gel electrophoresis and then transferred onto a PVDF membrane by Western blotting. The electroblotted proteins with N terminal acetylserine or acetylthreonine could be deblocked on-membrane by treatment with trifluoroacetic acid vapor and sequenced by a gas-phase protein sequencer. Similarly, N-formylated proteins could be deblocked on-membrane in HC1 solution and then directly sequenced from the N-terminal amino acid. Proteins with N-terminal pyroglutamic acid were enzymatically deblocked by in situ pyroglutamyl peptidase digestion, and N-acetylated proteins were also enzymatically deblocked with acylamino acid-releasing enzyme (AARE) after on membrane digestion with trypsin to generate the N-terminal peptide fragment. This tryptic digestion was required since AARE can remove the acetylamino acid only from a short peptide. Based on these four deblocking methods, we present a strategy for sequential deblocking and subsequent N-terminal sequence analysis of N-blocked protein immobilized on PVDF membrane. PMID- 8223392 TI - Electroblotting proteolytic products from native gel for direct N-terminal sequence analysis: an approach for studying protein-protein interaction. AB - Proteins which are electroblotted from native gels onto polyvinylidene difluoride (PVDF) membranes are suitable for detailed structural analysis. This method, in conjunction with limited proteolysis and N-terminal sequencing, has been used to study the molecular interactions between native protein molecules. The interaction between recombinant interleukin-2 (rIL-2) and its receptor (rIL-2R alpha) was examined as a model system. The working strategy consists of (i) proteolysis of rIL-2R alpha and rIL-2R alpha/rIL-2 complex, (ii) separation of the major proteolytic products by native polyacrylamide gel electrophoresis followed by electroblotting onto PVDF membrane, and (iii) sequence analysis of the blotted protein bands for the identification of peptide regions sensitive to proteolysis. Results have indicated that the exon 3 encoded region in rIL-2R alpha is sensitive to proteolysis regardless whether it is complexed with rIL-2 or not. This suggests that no major conformational changes occur in rIL-2R alpha during interaction with rIL-2. This electroblotting approach is, therefore, useful for studying protein-protein interaction in solution. PMID- 8223393 TI - Peptide dot immunoassay and immunoblotting: electroblotting from aluminum thin layer chromatography plates and isoelectric focusing gels to activated nitrocellulose. AB - Nitrocellulose membrane was preactivated with divinyl sulfone, and a spacer of 1,6-diaminohexane was coupled to the membrane which was functionalized by glutaraldehyde, leaving a reactive carbonyl group. The peptides were coupled to the carbonyl by the side chain and terminal amino groups. The octapeptide angiotensin II (sequence: DRVYIHPF) and peptide analogs containing 6-10 amino acid residues were dotted directly onto the matrix at 45 degrees C for 15 min and detected by specific antisera, which were raised in rabbits against angiotensin I and II, respectively. They were visualized by peroxidase-coupled anti-rabbit IgG antibodies. The detection limit for synthetic angiotensin II was 500 fg per cm2 (= 500 amol per cm2) and for the decapeptide angiotensin I (sequence: DRVYIHPFHL) it was 500 pg per cm2 (= 400 fmol per cm2). Separation of synthetic angiotensin analogs by high performance thin-layer chromatography on silica coated aluminum plates was followed by electroblotting onto activated nitrocellulose and detection with specific antibodies, showing a sensitivity of 100 fg and 1 pg for angiotensin II and angiotensin I, respectively. Isoelectric focusing in agarose using Ampholine carrier ampholytes and immunoblotting with specific antisera displayed a lower sensitivity for angiotensin II and angiotensin I of 2 ng and 20 ng, respectively. The isoelectric focusing and immunoblotting technique was applied for separation of angiotensin I and II and related peptides in serum, where synthetic angiotensin I was degraded in the presence of 1 mM phenylmethylsulfonyl fluoride and 10 mM ethylenediaminetetraacetic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223394 TI - Chemical activation of nitrocellulose membranes for peptide antigen-antibody binding studies: direct substitution of the nitrate group with diaminoalkane. AB - A method to covalently link peptide and proteins, through a diaminoalkane spacer to nitrocellulose membrane was developed for immunochemical applications. Initially the nitrocellulose membrane was modified by covalent incorporation of diaminoalkane spacers without any prior activation. The incorporation was shown primarily to involve alpha-elimination of the nitrate groups, and an imine was formed between the carbonyl group on the membrane and the diaminoalkane. The rate of incorporation increased exponentially with the length of the diaminoalkane as determined by a ninhydrin colorimetric reaction, which was developed for the study. More than 200 nmole diamine per mg nitrocellulose could be incorporated, but less than 11 nmol/mg (63 nmol/cm2) was chosen in order to retain the strength of the membrane. The primary amino groups of the modified membrane was glutaraldehyde activated and the octapeptide, angiotensin II, was covalently bound. A dot immunoassay was performed where specific anti-angiotensin II antibodies reacted with the peptide and was visualized by peroxidase coupled secondary antibodies. The results were quantified by video densitometry above 0.005 microgram AII per cm2. The immunoassay showed improved detection of the peptide on the activated as compared to unactivated membrane as well as increased retention of radiolabeled [125I]angiotensin II. PMID- 8223395 TI - Time-resolved fluorometry in nucleic acid hybridization and western blotting techniques. AB - Time-resolved fluorometry is now used extensively for immunological assays and to a lesser extent in other research areas. In this review I describe applications of time-resolved fluorometry in nucleic acid hybridization and in blotting techniques, including Southern and Western blotting. Clearly, time-resolved fluorometry has potential for playing a major role in techniques other than immunological assays. PMID- 8223396 TI - Western blotting of basic proteins after nondenaturing electrophoresis in acid conditions using the PhastSystem. AB - Electroblotting of basic proteins was performed from minigels after electrophoresis, under nondenaturing acidic conditions, by using the automated PhastSystem. Depending on the molecular masses of the proteins to be studied, various precast gel media were chosen. The transfer membranes with various types (nitrocellulose and polyvinylidene difluoride) and pore sizes (0.45 and 0.2 micron) were chosen accordingly. For the semidry electric transfer, a simple, discontinuous two-buffer system was used. The anode solution contained 0.3 M Tris, pH 10.4, and the cathode solution, 40 mM 6-amino-n-hexanoic acid, pH 7.6, with 20% v/v methanol each. The addition of 0.1% sodium dodecyl sulfate (SDS) in the cathode solution facilitated the elution of proteins from the gels and directed the migration of the negative SDS-protein complexes towards the anode membranes. The transfer conditions following native polyacrylamide gel electrophoresis allowed the visualization of basic proteins, with molecular weights ranging from 29,000 to 5,000, for which isoforms could be resolved and which retained their biological properties. PMID- 8223397 TI - Protein blotting analysis of large sample numbers. AB - A method is described that allows probing of hundreds of different antibodies on protein blots per day, making it possible to use protein blots for primary screening of hybridoma supernatants. It involves a miniaturization of the protein blots and their attachment to a solid support. This allows a great reduction of the required number of gels and electroblots and brings a drastic simplification of the incubation and washing procedures. Apart from a reduced resolution of closely adjacent bands the resulting mini-blots are comparable to conventional protein blots in sensitivity and specificity. The procedure has been successfully applied for primary screening of hybridoma supernatants. It can be useful in cases where a monoclonal antibody to an antigen with known molecular weight is required, or in situations where many different antibodies have to be tested in a short time period. PMID- 8223398 TI - Antibodies bound to nitrocellulose in acidic buffers retain biological activity. AB - This report compares the binding of proteins to nitrocellulose membranes in acidic buffers (pH 2 and 3) with binding in neutral buffer (pH 7). Initially, similar amounts of antibodies and other proteins bound to the nitrocellulose membrane in both acidic and neutral buffers. However, the susceptibility of individual proteins to displacement (stripping) from the membrane by the milk blocking agent depended on the pH of the buffer used to bind the proteins to the membrane. Most proteins that bound to nitrocellulose in acidic buffers were relatively resistant to milk-stripping compared to proteins bound in pH 7 buffer. Acid-binding of proteins to nitrocellulose also decreased the amount of protein that was stripped from the nitrocellulose membrane when Tween 20 was included in the washing buffer. After correcting for the amount of antibody remaining on the membrane after the milk block, it was found that acid-bound antibodies were unchanged in biological activity when compared with the same antibodies bound at neutral pH. These results suggest that acid-binding of proteins could increase the sensitivity of nitrocellulose membrane assays that use milk and/or Tween 20. PMID- 8223399 TI - Rapid electroelution of two-dimensionally separated protein mixtures: its use in in vitro assays of T cell activities. AB - A recently developed electroelution method for separated mixtures of proteins and its application in vaccine research were investigated. The method combines the high resolution power of two-dimensional gel electrophoresis with the advantage of direct probing of separated proteins with viable cells. An electroelution time of only 30 min was sufficient for complete protein transfer, as shown by Coomassie Brilliant Blue and silver staining. Inclusion of sodium dodecyl sulfate (SDS) into the electrophoresis buffer for the second dimension considerably improved the separation capacity. Furthermore, because of the low concentration of SDS (0.03%) no deleterious effects on the cells were seen. It was shown that T lymphocytes from cattle vaccinated with dead M. bovis BCG responded to numerous mycobacterial protein antigens, whereas unvaccinated control animals showed no, or very weak, responses. A comparison of T cell proliferation profiles obtained with different protein separations demonstrated the reproducibility of the method. PMID- 8223400 TI - Protein blot assays specific for the discrimination of the centromere autoantigen, CENP-A, from human cells. AB - The Western or protein blot has proven to be a valuable resource in detecting discrete, immunoreactive antigen targets associated with a variety of autoimmune diseases. As the roster of autoantigens has expanded, it has become increasingly common to tailor specific gel or blot conditions to a particular polypeptide antigen. Two such assays are reported here as applied to the fractionation and visualization of human centromere protein (CENP-A), a centromere autoantigen associated with the rheumatic disease, systemic sclerosis. The centromere antigens are effectively solubilized in the presence of 1 M MgCl2 to allow for further purification. CENP-A copurifies with the histone proteins, primarily H3 and H4. The two CENP-A-specific protein blot assays separate CENP-A from the histone proteins and enhance CENP-A immunoreactivity. The first assay is based on the use of acid-urea gels with a Triton X-100 concentration chosen to maximize separation of CENP-A from all the histones. The second assay is based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis to differentiate two very basic proteins of similar molecular weight, namely CENP-A and histone H3. For each gel system, a selective choice of associated immunoblot parameters allows for the reproducible discrimination of the CENP-A antigen. PMID- 8223401 TI - Immunoaffinity analysis of cross-reacting allergens by protein blotting. AB - IgE antibodies from sera having reactivity against ryegrass pollen protein allergens, wheat endosperm protein allergens and also several other cereal protein allergens were adsorbed with either ryegrass pollen or the wheat/globulin fraction immobilised on solid phases and subsequently eluted with low pH buffer. The eluted antibodies were reacted with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) blots of the different allergens. Antibodies adsorbed and subsequently eluted from the two allergen sources recognised different spectra of proteins in the ryegrass pollen and cereal allergen sources and indicated the degree of immunological cross-reactivity. Intra-species cross reactivity of IgE antibodies was demonstrated employing similar methods to those used for the pollen and cereal allergens by using a recombinant allergen from the venom of the ant Myrmecia pilosula as the immunoadsorbent protein on the solid phase. PMID- 8223402 TI - Intra-species cross-reactivity of house dust mite allergens separated by protein blotting and detected by selective elution of mite components and IgE antibodies. AB - A complex and varied antibody response to allergens, in particular Dermatophagoides pteronyssinus, is observed in the serum of allergic patients. To examine the role of cross-reacting antigenic determinants in allergy, a strategy has been devised for isolating antibody populations that bind to individual polypeptides of an allergen source separated by sodium dodecyl sulfate polyacrylamide electrophoresis and electroblotted to nitrocellulose. Polypeptides are identified on the blots, desorbed and coupled to CNBr-activated paper discs. The coupled discs are then used as immunoadsorbent surfaces for the binding of antibodies from allergic sera. Antibodies eluted from the polypeptide immunoadsorbents are then used to reprobe protein blots of the allergen. Results are presented which demonstrate strong cross-reaction between two major mite allergens, Der p I and Der p III, and some weaker intra-species cross reactivities. PMID- 8223403 TI - A study of the advantages and limitations of immunoblotting procedures for the detection of antibodies against influenza virus. AB - An immunoblotting procedure was used to determine the specificity and examine some of the properties of antibodies produced following infection of mice with influenza virus or inoculation with noninfectious material with Alhydrogel or complete Freund's adjuvant. The noninfectious material used was beta propiolactone-inactivated influenza virus and a preparation (HANA) enriched for the surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). When influenza viral proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions, each of the anti viral antisera tested exhibited strong binding. Under reducing conditions, however, much weaker binding was observed especially towards the HA1 subunit of HA. This was particularly apparent with antisera raised to virus or HANA in the absence of adjuvant. A panel of monoclonal antibodies directed to HA also bound well to viral HA separated by SDS-PAGE under nonreducing conditions but failed to recognize epitopes on HA1 separated under reducing conditions. These results suggest that when HA is reduced and immobilized on a solid support, it does not display the conformational features essential for the integrity of all epitopes. The immunoblotting procedure was also used to determine the isotype of anti-viral antibody directed against individual viral proteins and to detect matrix protein 2 (M2) in purified influenza virions and influenza-infected cells using antisera raised to a synthetic peptide representing a sequence within the M2 protein. PMID- 8223404 TI - Western blot as a tool in the diagnosis of Lyme borreliosis. AB - Borrelia burgdorferi is the causative agent of Lyme borreliosis, a multisystem disorder, which can mimic numerous immune disorders and inflammatory diseases. Laboratory diagnosis of Borrelia infection relies on immunodiagnostic assays, which, however, are hampered by unsatisfactory specificity. The Western blot technique has been employed to analyze the humoral immune response in Lyme borreliosis and is used as a serodiagnostic confirmation test. The most important immunodominant proteins of Borrelia burgdorferi are the 94 kDa, 60 kDa, 41 kDa (flagellin), 34 kDa (Osp B), 31 kDa (Osp A), 30 kDa, 21 kDa (Osp C), and 17/18 kDa proteins. Whereas the 60 kDa, 41 kDa, and 34 kDa constituents reveal a marked cross-antigenicity with other spirochetes and even more distantly related bacteria, antibodies against the 94 kDa, 31 kDa and 21 kDa proteins are largely species-specific. The early immune response in Lyme borreliosis is triggered mainly by the flagellin. In the later stage a wide range of immunogenic proteins is involved, with the 94 kDa antigen being the best marker for late immune response. If the Western blot is used for diagnostic purposes the differences between early and late-stage immunogenicity of Borrelia proteins must be taken into account. Interpretation criteria for blot positivity in early-stage borreliosis are primarily based on the presence of the 21 kDa band and the semiquantitatively recorded intensity of the 41 kDa band. In the diagnosis of late-stage infection, blot positivity relies on the presence of the 94 kDa, 39 kDa, 31 kDa, 30 kDa and 21 kDa bands. PMID- 8223405 TI - Humoral immune response against Helicobacter pylori as determined by immunoblot. AB - An immunoblot method has been evaluated to diagnose Helicobacter pylori infection serologically by comparing 69 serum specimens from patients with a positive Gram stain and/or culture result and a positive urease test on biopsy material, as well as 51 serum specimens from patients with at least 4 negative urease tests, and negative microscopy and culture results. Sensitivity and specificity was found to be 100%. Recognition of the cross-reacting flagellin (66 kDa), flagellar sheath protein (51 kDa), and a 14 kDa protein are not a criterion for a current H. pylori infection. On the other hand, any combination of at least two of the 180, 120, 90, 75, 67, 29.5 and 19 kDa bands were diagnostic of infection. Three H. pylori strains, which were compared with both gel electrophoretic analyses and immunoblot reactivity, exhibited in part strong qualitative and quantitative differences that particularly affect the 120 kDa pathogenic factor, the large urease subunit and other proteins especially in the molecular mass range from 50 to 67 kDa. IgG immunoblot patterns showed that the choice of H. pylori strain, as well as a reproducible and standardizable antigen preparation, is of great importance for the reliability of serodiagnostic tests. The immunoblot method was found to be a valuable tool for the semi-quantitative confirmation of results achieved with other serological methods as well as optimization and quality control of the antigens used for serodiagnostic purposes. PMID- 8223406 TI - Immunoblotting of Yersinia plasmid-encoded released proteins: a tool for serodiagnosis. AB - The plasmid-encoded, released proteins (RPs) of Yersinia enterocolitica serotypes 09 and 03 were separated by sodium dodecyl sulfate (SDS)-pore gradient gel electrophoresis. The RP-patterns of both serotypes proved to be identical. Five major proteins of M(r) 27,000, 34,700, 35,600, 45,800, and 46,800 were detected. Spontaneously plasmid-cured derivatives of the two serotypes lost the feature of protein release. By immunoblotting of RP with sera from patients suffering from acute Yersinia infections, specific and reproducible band patterns were obtained. Laser scan densitometry was applied to record the immunoreactions quantitatively. Predominant bands were detected at an M(r) of 34,700 and 35,600. IgA and IgM antibodies appeared as acute-phase markers rapidly decreasing in the reconvalescent phase. In contrast, immunoblots of patients with supposed chronic yersiniosis were characterized by a persisting IgA and elevated IgG reactivity. The application of RP as diagnostic antigens proved to be advantageous because they are naturally separated from cross-reacting proteins, common to pathogenic and nonpathogenic strains of Yersinia enterocolitica and Y. pseudotuberculosis. PMID- 8223407 TI - Trunk muscle activation and cocontraction while resisting applied moments in a twisted posture. AB - Previous studies of twisting have revealed substantial cocontraction of agonist and antagonist muscles within the torso when torsional moments are generated. The objective of the current study was to quantify the activations and cocontraction of eight trunk muscles as subjects maintained an axially rotated trunk posture and resisted external applied bending moments. Ten subjects twisted their torsos 25 degrees to the right (clockwise) and resisted 20 and 40 Nm bending moments from 12 directions. The moment directions were in a transverse plane and labelled clockwise as viewed from above, ranging from 0 degrees (mid-saggital, anterior) to 330 degrees, in 30 degrees increments. RMS EMG amplitude data were collected using surface electrodes and normalized to maximal voluntary contractions. Significant changes were observed in the muscle responses due to the interaction of the moment direction and moment magnitude for six of the eight muscles tested. Comparison of the present data with that collected previously in neutral postures indicated: (1) a large increase in the activation levels of the right erector spinae and the left external oblique muscles; and (2) a counter-clockwise shift in the moment direction at which the peak activation of these same muscles occurs. Analysis of the relative activation levels (RALs), constructed from the NEMG data to quantify the cocontraction, indicated that the changes in cocontraction were more robust in response to changes in the bending moment's direction as opposed to changes in bending moment's magnitude. PMID- 8223408 TI - Audibility and identification of auditory alarms in the operating room and intensive care unit. AB - The audibility and the identification of 23 auditory alarms in the intensive care unit (ICU) and 26 auditory alarms in the operating rooms (ORs) of a 214-bed Canadian teaching hospital were investigated. Digital tape recordings of the alarms were made and analysed using masked-threshold software developed at the Universite de Montreal. The digital recordings were also presented to the hospital personnel responsible for monitoring these alarms on an individual basis in order to determine how many of the alarms they would be able to identify when they heard them. Several of the alarms in both areas of the hospital could mask other alarms in the same area, and many of the alarms in the operating rooms could be masked by the sound of a surgical saw or a surgical drill. The staff in the OR (anaesthetists, anaesthesia residents, and OR technologists) were able to identify a mean of between 10 and 15 of the 26 alarms found in their operating theatres. The ICU nurses were able to identify a mean of between 9 and 14 of the 23 alarms found in their ICU. Alarm importance was positively correlated with the frequency of alarm identification in the case of the OR, rho = 0.411, but was not significantly correlated in the case of the ICU, rho = 0.155. This study demonstrates the poor design of auditory warning signals in hospitals and the need for standardization of alarms on medical equipment. PMID- 8223409 TI - Performance and alertness on 8 h and 12 h rotating shifts at a natural gas utility. AB - An 8 h/5-7 day shift schedule was compared with a newly instituted 12 h/2-4 day schedule in this, our second worksite study of extended workshifts. Workers completed a performance/alertness test battery, and a questionnaire on sleep patterns and other personal habits, 2-4 times a week on all shifts. After 10 months adaptation to the 12 h shift schedule, there were decrements in performance/alertness attributable to the extra 4 h on the extended shift. There were also reductions in sleep across the workweek which were most apparent on 12 h night shifts. The results are consistent with our first worksite study of 12 h shifts and indicate extra caution should be exercised when scheduling critical activities for extended workshifts, especially extended night shifts. PMID- 8223410 TI - Effects of graded dosages of alcohol on nine computerized repeated-measures tests. AB - We report a controlled laboratory validation experiment to provide an empirical sequel to the general background of the development of the Automated Performance Test System (APTS) that was presented in Turnage et al. (1992). The purpose of this study was to index performance deficit against various Blood Alcohol Concentrations (BACs) of small (0.05% BAC), medium (0.10% BAC), and large (0.15% BAC) dosages as well as a placebo (0.00 BAC) condition. Blood alcohol concentrations, measured in four different ways, were highly reliable and the method with the highest intercorrelation was whole blood. When taken singly, eight of the nine tests produced significant (p < 0.001) relationships with the disparate blood alcohol levels, which were essentially monotonic. A multiple regression analysis suggested that most of the tests were behaving similarly and that two or three tests accounted for 52 to 54% of the variance attributable to alcohol that was explained by the longer battery. While all tests appear valid, some of them appeared more sensitive than others (viz., code substitution, manikin, and choice reaction time). From the standpoint of these tests, greater changes occurred in cognitive function between the placebo and 0.05 level than between the 0.05 and 0.10 levels. However, the greatest reduction in performance occurred between 0.10 and 0.15, and the relatively abrupt nature of this change implies that sharp decrements in cognitive performance occur at that point. There were individual differences in resistance to alcohol, and there is strong inference that these differences would be reliable if they were tested again. We believe that further development and study of such techniques is warranted for use in fitness-for-duty testing and development of a dose equivalency index. PMID- 8223411 TI - Vapour transfer in two-layer clothing due to diffusion and ventilation. AB - An experiment was carried out to measure the vapour resistance of two-layer clothing ensembles as a function of air permeability of the outer layer, open or closed apertures, wind, and walking, both for the total ensemble and for the outer garment alone. Six subjects walked on a treadmill (0.0, 2.5, and 5 km.h-1) which was placed in a wind tunnel (0.2, 0.7, and 3.0 m.s-1). They wore long underwear and an outer garment made of impermeable (imp), microporous (mpo), low air permeable (loa), or high air permeable (hia) fabric. Vapour resistances were determined by a trace gas method, calibrated against water vapour resistance. The vapour resistances of the underclothing and the outer garment were calculated from the measured data, as was the ventilation through the apertures. The vapour resistance of the underclothing was almost constant at 5 mm air equivalent. The ventilation was strongly dependent on wind and motion but still so low (54 l.min 1) that only the impermeable garment could benefit from it noticeably. The vapour resistance of the garments also varied strongly (imp 55-200 mm, mpo 12-20 mm, loa and hia 1-14 mm air equivalent). For the imp garment, this is due to leakage of air, whereas the vapour permeable garments were dominated by the diffusion and air penetration through the fabric. It is concluded that ventilation with vents cannot match the effect of vapour permeability, and that real low vapour resistances are only possible with air permeable fabrics. PMID- 8223412 TI - Physiological strains in hot-humid conditions while wearing disposable protective clothing commonly used by the asbestos removal industry. AB - The purpose of this study is to investigate workers' responses to work in hot humid conditions while wearing protective clothing commonly used by the asbestos removal industry, and to evaluate the effects of resting between work bouts in a cool environment on the physiological strain. Seven male students wearing impermeable protective clothing and air masks were exposed to the following conditions for 100 min on separate days: (1) hot conditions (35 degrees C/85%RH), (2) cool conditions (20 degrees C/85%RH), and (3) hot/cool conditions (working in hot conditions and resting in cool conditions). After 12 min rest, the subjects worked on an ergometer (70 Watts) for 18 min. This experimental schedule was repeated three times under each environmental condition. Rectal temperature (Tre), heart rate (HR), sweat rate (SR) and discomfort sensation were recorded. Two of the subjects were not able to complete the experiment in hot conditions. The increases in Tre and HR with time were not found in cool conditions. Although Tre increased in hot/cool conditions, it was almost half of that in hot conditions. Since HR did not return to the pre-work level during recovery in hot conditions, HR during work was accompanied by increases in HR at pre-work. HR during work in hot/cool conditions was higher than that in cool conditions, HR at pre-work, however, was almost the same as that in cool conditions because of rapid recovery. The means of SR in hot and hot/cool conditions were five and four times greater than that in cool conditions, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223413 TI - The brainstem cardioarrhythmogenic triggers and their possible role in sudden epileptic death. AB - The cardiovascular effects of simultaneous activation of hypothalamic and mesencephalic cardioarrhythmogenic triggers were studied in hemispherectomized rats. Paroxysmal activity of hypothalamic neurons (HEF), elicited by topical application of penicillin G on the thalamus, triggered short-lasting bradyarrhythmic episodes, up to a maximum of 6 s, and alterations in repolarization. In the hypothalamic neurons, an additional penicillin G epileptic focus at mesencephalic level (MEF) induced the enhancement of paroxysmal activity by a recruitment of new units and potentiation of their background activity. HEF+MEF triggered second-degree 2:1-8:1 atrioventricular (A-V) blocks, impairment of the A-V conduction, alterations in the recovery phase and bundle branch blocks. After HEF, the arterial blood pressure decreased by 4-6%. HEF+MEF induced a further reduction of 17% in systolic pressure only. It is possible that the enhancement of the HEF following MEF could depend on MEF spreading upward. The HEF, in turn, by spreading downward could influence the MEF and so activate, between HEF and MEF, a circuitry with reciprocal co-excitation that could explain the more serious cardiovascular alterations observed during HEF+MEF compared with those observed during HEF only or during MEF only. However, this cardiovascular impairment, which must be neurogenic in origin as it was observed in animals with normal acid-base and blood parameter values, did not induce heart death. Thus, additional concomitances must be considered, such as metabolic derangement which can occur during seizures, to explain sudden death in epileptic patients. Some aspects of metabolic complications in cardiac activity during epilepsy are also discussed. PMID- 8223414 TI - The influence of thalamic GABA transmission on the susceptibility of adult rats to flurothyl induced seizures. AB - There is considerable evidence that thalamic nuclei are involved in the propagation and regulation of seizures. In the present study, we investigated the possible role in seizure mechanisms of GABAergic transmission in two thalamic nuclei, the posterior nucleus (PO) and the ventromedial nucleus (VM). Several GABAergic drugs were bilaterally microinfused into PO or VM of adult rats via chronically implanted cannulae, before testing the rats' susceptibility to seizures induced by flurothyl. In PO, infusions of the GABA elevating agent gamma vinyl-GABA (20 micrograms) or of the GABAA receptor agonist muscimol (100 ng) suppressed both clonic and tonic seizures. Infusions into PO of the GABAA receptor antagonist bicuculline (100 ng) facilitated both these seizure types. Administration of the GABAB receptor agonist baclofen (200 ng) also suppressed clonic seizure susceptibility. Drug infusions into VM, however, did not significantly modify the susceptibility to seizures. These findings lead us to conclude that GABAergic transmission in the vicinity of the PO, but perhaps not in VM, affects flurothyl seizure susceptibility. We hypothesize that GABA synapses in PO may be part of a seizure propagation or control circuit including striatum, substantia nigra, and superior colliculus. PMID- 8223415 TI - Acute and chronic anticonvulsant effects of D(-)CPPene in genetically epilepsy prone rats. AB - Acute administration (0.4-12 mumol/kg, i.p.) of the NMDA antagonist D(-)CPPene (3 (2-carboxypiperazin-4-yl)propenyl-1-phosphonic acid; molecular weight = 250.2) dose-dependently reduced the incidence of sound-induced clonic seizures in genetically epilepsy-prone (GEP) rats. The ED50 value against clonic seizure at +1 h was 2.6 (2.2-3.2) and at +4 h was 1.7 (1.3-2.3) mumol/kg (i.p.). Twelve hours after chronic administration of an anticonvulsant dose of D(-)CPPene (4 mumol/kg, i.p., twice daily at 09.00 and 21.00 h, for 14 days), repeat administration of D(-)CPPene (1.2-12 mumol/kg, i.p.) gave ED50 values against clonic seizure at +1 h of 3.4 (2.5-4.8) mumol/kg and at +4 h of 2.8 (2.0-3.9) mumol/kg (i.p.). There was no significant difference observed between ED50 values for these acute and chronic groups. The duration of the anticonvulsant effect observed between +1 h and +8 h after D(-)CPPene (0.4-12 mumol/kg, i.p.) was similar in the acute and chronic groups. The ED50 values for D(-)CPPene-induced impairment of locomotor performance (using a rotarod) at +3 h were 6.8 (4.4-10.4) and 6.4 (4.1-10.1) mumol/kg (i.p.) for the acute and chronic groups respectively. At +7 h after D(-)CPPene administration the ED50 value for locomotor impairment was 19.5 (10.6-36.0) mumol/kg (i.p.) in the acute group. In this study there is no evidence of tolerance to the anticonvulsant effects and little evidence of tolerance to the adverse effects (ataxia) of D(-)CPPene in GEPRs. PMID- 8223416 TI - Mechanisms of kindling: an evaluation of single trial seizure induction procedures for use as controls. AB - Investigations into the neurochemical or molecular biological mechanisms underlying the kindled state require a seizure induction procedure for eliciting generalized tonic-clonic seizures in naive animals. Such seizure controls are necessary for dissociating the influence of ictal motor events on measures of interest from the influence of the kindling process on these same measures. In this study three procedures for inducing seizures in naive animals were evaluated against a set of criteria considered ideal. PMID- 8223417 TI - Pharmacological characterization of phenytoin-resistant amygdala-kindled rats, a new model of drug-resistant partial epilepsy. AB - By repeated treatment with phenytoin, subgroups of animals with different response to phenytoin were selected from a large group of amygdala-kindled rats. In one subgroup ('phenytoin responders') phenytoin induced reproducible increases in focal seizure threshold, while in another subgroup ('phenytoin nonresponders') animals were resistant to phenytoin. These phenytoin nonresponders, which comprised about 12% of the kindled animals tested, did not differ from phenytoin responders in drug levels, drug adverse effects or location of the stimulation electrode in the amygdala. Treatment of phenytoin responders and nonresponders with other primary antiepileptic drugs showed that valproate and phenobarbital induced much smaller increases in focal seizure threshold in phenytoin nonresponders than in responders, whereas carbamazepine induced about the same threshold increase in both groups. The novel antiepileptic drug vigabatrin, which acts by increasing GABA levels, exerted anticonvulsant effects in phenytoin responders but was inactive in nonresponders. Determination of plasma amino acids before and after vigabatrin treatment demonstrated marked differences in biochemical responses to vigabatrin although drug levels were about the same in both groups. The data demonstrate that amygdala-kindled rats with phenytoin resistance offer a basic approach to the investigation of mechanisms of drug resistance in epilepsy. Furthermore, these animals may be used in the evaluation of new anticonvulsant drugs for treatment of partial seizures which do not respond to the currently available therapies. PMID- 8223418 TI - Effects of single and repeated administration of vigabatrin on the performance of non-epileptic rats in a delayed non-matching to position task. AB - The present experiments were performed to investigate the effects of single and repeated administration of vigabatrin (gamma-vinyl-GABA), a novel antiepileptic drug, on a working memory task (delayed non-matching to position task) in non epileptic rats. At doses of 100, 300 and 500 mg/kg single administrations of vigabatrin and 50, 100 and 200 mg/kg repeated administrations, vigabatrin did not affect the choice accuracy in the delayed non-matching to position task employing delays of 0, 1, 2, 4, 8 and 16 s, whereas repeated administration with 300 mg/kg and a single dose of 1000 mg/kg decreased the behavioral activity as compared to saline treatment. Previous studies have shown that at doses of 50-200 mg/kg (daily administration) and 200-1000 mg/kg (single administration) vigabatrin has anticonvulsant activity. The present results suggest that vigabatrin does not markedly impair working memory in the low range of antiepileptic/anticonvulsive doses. PMID- 8223419 TI - GABAA receptor impairment in the genetic absence epilepsy rats from Strasbourg (GAERS): an immunocytochemical and receptor binding autoradiographic study. AB - Some aspects of the GABA and cholinergic systems have been investigated in the cortex and thalamus of GAERS Wistar rats, a model of petit-mal epilepsy, and in a non-epileptic control strain. GABA and its synthetic enzyme, glutamic acid decarboxylase (GAD), were located by immunocytochemistry; the GABAA receptors were evaluated by autoradiography of GABA-enhanced 3H-flunitrazepam binding and by immunocytochemistry using specific antibodies against the beta 2-beta 3 subunits of GABAA receptor protein. GABA and GAD immunocytochemistry did not show up any difference in density or distribution of immunoreactive elements (fibers, terminals and neurons) between epileptic and control animals, but autoradiographic and immunocytochemical studies showed a decreased enhancement of 3H-flunitrazepam binding and of beta 2-beta 3 subunits of GABAA receptor in the sensorimotor cortex and anterior thalamic areas of the epileptic strain. No differences were found in benzodiazepine receptors in the two strains. GABAB receptors were measured as 3H-baclofen binding in a crude synaptic membrane preparation and there was no difference between epileptic and control animals. Choline acetyltransferase, the synthetic enzyme for acetylcholine, and muscarinic receptor subtypes (M1 and M2), visualized respectively by an immunocytochemical procedure and binding autoradiography, did not differ in epileptic and normal rats. The data suggest an impairment of the 'GABAA system' in restricted brain regions of epileptic rats, due to a reduction of receptor beta 2-beta 3 subunits and coupling to benzodiazepine receptors despite the normal synthesis and location of the neurotransmitter. PMID- 8223420 TI - Intracarotid amobarbital testing for language and memory dominance in children. AB - The intracarotid amobarbital procedure (IAP) was attempted in 22 pediatric epilepsy surgery candidates, ages 5-12 years old. With extra pre-test teaching and emotional preparation, adjusted amobarbital dosage for younger patients, and simplified test items tailored to the child's abilities, language and memory testing were accomplished after at least one injection for 19 (86%) of patients. Language dominance was clarified in 11 children overall (50% of patients), in all of the children who had bilateral testing and at least borderline intelligence (IQ > 70), and in 57% of the children with mild or moderate mental retardation. Memory assessment was accomplished in 18 children after amobarbital injection of the hemisphere ipsilateral to the predominant epileptogenic zone. IAP retention scores tended to be lower in children than adults, especially in the setting of mental retardation. Retention scores after ipsilateral injection were > 60% in all 10 of the children with at least borderline intelligence, but < 60% (25-50%) in five of eight children with mental retardation. Retention scores after contralateral injection were > 60% in only four of 12 children. Our results suggested that a modified pediatric IAP protocol can clarify the hemisphere of language dominance in most verbal preadolescent children with at least borderline intelligence, and in many children with mental retardation. However, IAP memory retention scores tended to be lower in children than adults and should be interpreted with caution. PMID- 8223421 TI - Quantitative MRI hippocampal volumes: association with onset and duration of epilepsy, and febrile convulsions in temporal lobectomy patients. AB - The relationships between preoperatively acquired MRI-based hippocampal volumes (HV), seizure disorder onset and duration, and early childhood febrile convulsions were investigated retrospectively with data from 72 left and 56 right temporal lobectomy patients. Patients with lesional pathology and heterotopic abnormalities were excluded. Age at development of spontaneous seizures unprovoked by an acute illness defined age of seizure disorder onset. Age of onset was subtracted from age at neurosurgery to determine duration. MRI variables included in this study were the right and left HV divided by total intracranial volume (RAHV, LAHV), and the right-left hippocampal difference (DHF). Partial correlations were used to better isolate relationships with onset of recurrent seizures corrected for age at surgery, and age at neurosurgery corrected for age of recurrent seizure onset. Partial correlations between age at neurosurgery and volume were not significant in either group. LAHV (r = 0.42, P < 0.0003) and DHF (r = -0.49, P < 0.0001) were correlated with age of onset in the left lobectomy group. Correlations in the right lobectomy group were not significant. The presence of a febrile convulsion was associated with smaller LAHV (F(1,70) = 10.54, P < 0.002) and larger DHF (F(1,70) = 11.36, P < 0.002) in left temporal lobectomy patients. The presence of a febrile convulsion in the right temporal group was associated with a slightly smaller DHF (F(1,56) = 5.90, P < 0.02), and slightly smaller RAHV (F(1,56) = 4.49, P < 0.04). These data suggest that hippocampal atrophy remains stable over the duration of temporal lobe onset seizure disorders, and is associated with early onset of recurrent seizures in left temporal patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223422 TI - Evaluation of the thiopental test in epilepsy surgery patients. AB - One hundred and three patients underwent a thiopental test during a presurgical evaluation for epilepsy surgery. Depth electrodes were implanted bilaterally into the mesial temporal and mesial frontal lobes. Thiopental was infused at 25 mg/30 s for adults and 0.3 mg/kg/20 s in children until loss of corneal reflexes or a total of 1 g. The absence of beta activity on the EEG, and activation of interictal spiking were two parameters monitored at all recording sites. Positive results were correlated with the anatomy of the ictal epileptic foci. Half of the patients produced increased interictal spiking and three-quarters of them showed absence of beta production in at least one lobe. Despite a reasonably high concordance between ictal foci and the two thiopental parameters, both measures yielded a low sensitivity and specificity. This work, as well as that reported in the literature, was unable to confirm the hypothesis that local damage within the temporal lobe was responsible for positive thiopental test results. An alternative hypothesis is proposed that the thalamus may be a distant source of these findings. Irrespective of the mechanism, the thiopental test must be used cautiously in the evaluation for epilepsy surgery. PMID- 8223423 TI - Molecular genetic analysis of a locus required for resistance to antimicrobial peptides in Salmonella typhimurium. AB - The innate immunity of vertebrates and invertebrates to microbial infection is mediated in part by small cationic peptides with antimicrobial activity. Successful pathogens have evolved mechanisms to withstand the antibiotic activity of these molecules. We have isolated a set of genes from Salmonella typhimurium which are required for virulence and resistance to the antimicrobial peptides melittin and protamine. Sequence analysis of a 5.7 kb segment from the wild-type plasmid conferring resistance to protamine contained five open reading frames: sapA, sapB, sapC, sapD and sapF, organized in an operon structure and transcribed as a 5.3 kb mRNA. SapD and SapF exhibited similarity with the 'ATP binding cassette' family of transporters including the bacterial Opp and SpoOK, involved in the uptake of oligopeptides; the yeast STE6, necessary for the export of a peptide pheromone; and the mammalian mdr, which mediates resistance to chemotherapeutic agents in cancer cells. SapA showed identity with other periplasmic solute binding proteins involved in peptide transport. The SapABCDF system constitutes a novel transporter for enteric bacteria and the first one harboring a periplasmic component with a role in virulence. PMID- 8223424 TI - The Sec13p complex and reconstitution of vesicle budding from the ER with purified cytosolic proteins. AB - SEC13 encodes a 33 kDa protein that participates in vesicle budding from the endoplasmic reticulum (ER). In order to purify a functional form of Sec13p, a SEC13-dihydrofolate reductase (mouse) fusion gene (SEC13:DHFR) was constructed that complements both sec13 temperature sensitive and null mutations. Methotrexate-agarose affinity chromatography facilitated the purification of two forms of the Sec13-dhfrp fusion protein: a monomeric form and a high molecular weight complex. The complex form consists of two subunits: Sec13-dhfrp and a 150 kDa protein (p150). Native immunoprecipitation experiments confirm that Sec13p exists in a complex with p150 in wild type cells. Functional analysis supports a role for both subunits in protein transport. Vesicle budding from the ER in a cell-free reaction is inhibited by Fab antibody fragments directed against either Sec13p or p150. The purified Sec13-dhfrp/p150 complex, but not the Sec13-dhfrp monomer, in combination with two other pure protein fractions (Sar1p and a Sec23/Sec24 protein complex) satisfies the requirement for cytosol in a cell-free vesicle budding reaction. The vesicles formed with the purified protein fractions are competent to fuse with the Golgi and are biochemically distinct from the ER membrane fraction from which they derive. PMID- 8223425 TI - The yeast SSS1 gene is essential for secretory protein translocation and encodes a conserved protein of the endoplasmic reticulum. AB - The SEC61, SEC62 and SEC63 yeast gene products are membrane components of the apparatus that catalyses protein translocation into the endoplasmic reticulum (ER). In the hope of uncovering additional components of the translocation apparatus, we sought yeast genes whose overexpression would restore partial thermoresistance in a sec61 translocation-deficient mutant. The first extragenic Sec sixty-one suppressor, SSS1, is an essential single copy gene whose overexpression restores translocation in the sec61 mutant. Another extragenic suppressor was identified as TDH3, which encodes the major isozyme of the most abundant yeast protein, glyceraldehyde-3-phosphate dehydrogenase. TDH3 overexpression could exert an indirect effect by competitively inhibiting protein synthesis, thereby allowing the impaired translocation apparatus to cope with a reduced flow of newly synthesized secretory proteins. Depletion of the Sss1 protein rapidly results in accumulation of multiple secretory or membrane proteins devoid of post-translational modifications; the normally secreted alpha factor accumulates on the cytosolic side of ER membranes. Thus, the SSS1 gene is required for continued translocation of secretory preproteins beyond their early association to ER membranes. Consistent with its essential role in protein translocation, the Sss1 protein localizes to the ER and homologues were detected in higher eukaryotes. PMID- 8223426 TI - Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport. AB - The yeast SEC1 gene encodes a hydrophilic protein that functions at the terminal stage in secretion. We have cloned two yeast genes, SSO1 and SSO2, which in high copy number can suppress sec1 mutations and also mutations in several other late acting SEC genes, such as SEC3, SEC5, SEC9 and SEC15. SSO1 and SSO2 encode small proteins with N-terminal hydrophilic domains and C-terminal hydrophobic tails. The two proteins are 72% identical in sequence and together perform an essential function late in secretion. Sso1p and Sso2p show significant sequence similarity to six other proteins. Two of these, Sed5p and Pep12p, are yeast proteins that function in transport from ER to Golgi and from Golgi to the vacuole, respectively. Also related to Sso1p and Sso2p are three mammalian proteins: epimorphin, syntaxin A/HPC-1 and syntaxin B. A nematode cDNA product also belongs to the new protein family. The new protein family is thus present in a wide variety of eukaryotic cells, where its members function at different stages in vesicular transport. PMID- 8223427 TI - Multiple pathways for protein transport into or across the thylakoid membrane. AB - Many thylakoid proteins are cytosolically synthesized and have to cross the two chloroplast envelope membranes as well as the thylakoid membrane en route to their functional locations. In order to investigate the localization pathways of these proteins, we over-expressed precursor proteins in Escherichia coli and used them in competition studies. Competition was conducted for import into the chloroplast and for transport into or across isolated thylakoids. We also developed a novel in organello method whereby competition for thylakoid transport occurred within intact chloroplasts. Import of all precursors into chloroplasts was similarly inhibited by saturating concentrations of the precursor to the OE23 protein. In contrast, competition for thylakoid transport revealed three distinct precursor specificity groups. Lumen-resident proteins OE23 and OE17 constitute one group, lumenal proteins plastocyanin and OE33 a second, and the membrane protein LHCP a third. The specificity determined by competition correlates with previously determined protein-specific energy requirements for thylakoid transport. Taken together, these results suggest that thylakoid precursor proteins are imported into chloroplasts on a common import apparatus, whereupon they enter one of several precursor-specific thylakoid transport pathways. PMID- 8223428 TI - Functional cooperation of mitochondrial protein import receptors in yeast. AB - We have identified a 20 kDa yeast mitochondrial outer membrane protein (termed MAS20) which appears to function as a protein import receptor. We cloned, sequenced and physically mapped the MAS20 gene and found that the protein is homologous to the MOM19 import receptor from Neurospora crassa. MAS20 and MOM19 contain the sequence motif F-X-K-A-L-X-V/L, which is repeated several times with minor variations in the MAS70/MOM72 receptors. To determine how MAS20 functions together with the previously identified yeast receptor MAS70, we constructed yeast mutants lacking either one or both of the receptors. Deletion of either receptor alone had little or no effect on fermentative growth and only partially inhibited mitochondrial protein import in vivo. Deletion of both receptors was lethal. Deleting only MAS70 did not affect respiration; deleting only MAS20 caused loss of respiration, but respiration could be restored by overexpressing MAS70. Import of the F1-ATPase beta-subunit into isolated mitochondria was only partly inhibited by IgGs against either MAS20 or MAS70, but both IgGs inhibited import completely. We conclude that the two receptors have overlapping specificities for mitochondrial precursor proteins and that neither receptor is by itself essential. PMID- 8223429 TI - Roles of plant homologs of Rab1p and Rab7p in the biogenesis of the peribacteroid membrane, a subcellular compartment formed de novo during root nodule symbiosis. AB - The peribacteroid membrane (PBM) in legume root nodules is derived from plasma membrane following endocytosis of Rhizobium by fusion of newly synthesized vesicles. We studied the roles of plant Rab1p and Rab7p homologs, the small GTP binding proteins involved in vesicular transport, in the biogenesis of the PBM. Three cDNAs encoding legume homologs of mammalian Rab1p and Rab7p were isolated from soybean (sRab1p, sRab7p) and Vigna aconitifolia (vRab7p). sRab1p was confirmed to be a functional counterpart of yeast Ypt1p (Rab1p) by complementation of a yeast ypt1-1 mutant. Both srab1 and vrab7 genes are induced during nodulation with the level of vrab7 mRNA being 12 times higher than that in root meristem and leaves. This induction directly correlates with membrane proliferation in nodules. Antisense constructs of srab1 and vrab7, under a nodule specific promoter (leghemoglobin, Lbc3), were made in a binary vector and transgenic nodules were developed on soybean hairy roots obtained through Agrobacterium rhizogenes-mediated transformation. Both antisense srab1 and vrab7 nodules were smaller in size and showed lower nitrogenase activity than controls. The antisense srab1 nodules showed lack of expansion of infected cells, fewer bacteroids per cell and their frequent release into vacuoles. In contrast, antisense vrab7 expressing nodules showed accumulation of late endosomal structure and multivesicular bodies in the perinuclear region. These data suggest that both Rab1p and Rab7p are essential for the development of the PBM compartment in effective symbiosis. PMID- 8223430 TI - First structure of a snake venom metalloproteinase: a prototype for matrix metalloproteinases/collagenases. AB - Adamalysin II, a 24 kDa zinc endopeptidase from the snake venom of Crotalus adamanteus, is a member of a large family of metalloproteinases isolated as small proteinases or proteolytic domains of mosaic haemorrhagic proteins from various snake venoms. Homologous domains have recently been detected in multimodular mammalian reproductive tract proteins. The 2.0 A crystal structure of adamalysin II reveals an ellipsoidal molecule with a shallow active-site cleft separating a relatively irregularly folded subdomain from the calcium-binding main molecular body composed of a five-stranded beta-sheet and four alpha-helices. The folding of the peptide fragment containing the zinc-binding motif HExxHxxGxxH bears only a distant resemblance to thermolysin, but is identical to that found in astacin, with the three histidines and a water molecule (linked to the glutamic acid) likewise constituting the zinc ligand; adamalysin II lacks a fifth (tyrosine) zinc ligand, however, leaving its zinc ion tetrahedrally co-ordinated. Furthermore, adamalysin II and astacin share an identical active-site basement formed by a common Metturn. Due to their virtually identical active-site environment and similar folding topology, the snake venom metalloproteinases (hitherto called adamalysins) and the astacins (and presumably also the matrix metalloproteinases/mammalian collagenases and the Serratia proteinase-like large bacterial proteinases) might be grouped into a common superfamily with distinct differences from the thermolysin family. PMID- 8223431 TI - FGF3 from Xenopus laevis. AB - Fibroblast growth factor 3 (FGF3) was first identified as the product of a cellular oncogene activated by mouse mammary tumour virus but its normal role appears to be in the developing embryo. To gain further insights into its function, we have isolated sequences encoding the FGF3 homologue in Xenopus laevis, XFGF3. COS-1 cells transfected with XFGF3 cDNA express a 31 kDa product, p31, generated by signal peptide cleavage and Asn-linked glycosylation at the single consensus site. This product is secreted and becomes associated with the cell surface and extracellular matrix. Proteolytic cleavage of p31 in the extracellular compartment results in an amino-terminally truncated product, p27, that is also glycosylated. Both p31 and p27 bind quantitatively to heparin Sepharose and can be displaced from the cell surface and extracellular matrix by soluble heparin. Conditioned medium containing these two proteins is capable of inducing transient morphological transformation of NIH3T3 cells and of stimulating DNA synthesis in quiescent C57MG and BALB/MK cells which express different isoforms of FGF receptors 1 and 2. Since XFGF3 behaves very differently from its mouse counterpart, we constructed chimeras in which amino-terminal sequences from XFGF3 were fused with carboxy-terminal sequences from mouse FGF3. Increasing the contribution from mouse FGF3 led to a more restricted host range for the chimeric ligand. PMID- 8223432 TI - Definition of a novel ligand binding domain of a nuclear bHLH receptor: co localization of ligand and hsp90 binding activities within the regulable inactivation domain of the dioxin receptor. AB - The dioxin receptor mediates signal transduction by dioxin (2,3,7,8 tetrachlorodibenzo-p-dioxin) and binds to DNA target sequences as a heterodimer of the approximately 100 kDa ligand binding receptor and the approximately 85 kDa auxiliary factor, Arnt. Both of these factors encompass an N-terminal basic helix loop-helix (bHLH) motif required for DNA binding and dimerization. In this study we describe the construction of glucocorticoid/dioxin receptor fusion proteins which allow the regulation of glucocorticoid receptor activity by dioxin in transient transfections of CHO and hepatoma cells. Thus, in the absence of dioxin, chimeric receptor constructs which contain large 500-720 amino acid C terminal dioxin receptor fragments, but lack the N-terminal bHLH motif, confer repression upon the transcriptional activity of a glucocorticoid receptor derivative, tau DBD, containing its N-terminal strong transactivating signal (tau) and its DNA binding domain (DBD). In the presence of dioxin, this repression is reversed. Importantly, these chimeric receptors did not require the bHLH Arnt co-factor for function. A considerably smaller region of the dioxin receptor, located between amino acids 230 and 421, showed specific dioxin binding activity in vitro. Moreover, dioxin binding in vitro correlated with the ability of receptor fragments to form stable complexes in vitro with the molecular chaperone hsp90. These findings support the notion that hsp90 may be important for folding of a dioxin binding configuration of the receptor. Finally, tau DBD activity was constitutively repressed in a dioxin non-responsive manner by dioxin receptor fragments which failed to bind ligand but also failed to bind hsp90 in vitro, indicating that alternative mechanisms in addition to hsp90 binding may contribute to the inactivation function. In summary, the dioxin receptor system provides a novel and complex model of regulation of bHLH factors that may also give important insights into the mechanism of action of ligand-activated nuclear receptors. PMID- 8223433 TI - Signal transduction by the high-affinity GM-CSF receptor: two distinct cytoplasmic regions of the common beta subunit responsible for different signaling. AB - The high-affinity receptors for granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin 3 (IL-3) and IL-5 consist of two subunits, alpha and beta. The alpha subunits are specific to each cytokine and the same beta subunit (beta c) is shared by these three receptors. Although none of these receptor subunits has intrinsic kinase activity, these cytokines induce protein tyrosine phosphorylation, activation of Ras, Raf-1 and MAP kinase, and transcriptional activation of nuclear proto-oncogenes such as c-myc, c-fos and c-jun. In this paper, we describe a detailed analysis of the signaling potential of the beta c subunit by using a series of cytoplasmic deletion mutants. The human beta c consists of 881 amino acid residues. A C-terminal deletion mutant of beta c at amino acid 763 (beta 763) induced phosphorylation of Shc and activation of Ras, Raf-1, MAP kinase and p70 S6 kinase, whereas a deletion at amino acid 626 (beta 626) induced none of these effects. The beta 763 mutant, as well as the full length beta c, induced transcription of c-myc, c-fos and c-jun. Deletions at amino acid 517 (beta 517) and 626 (beta 626) induced c-myc and pim-1, but no induction of c-fos and c-jun was observed. GM-CSF increased phosphatidylinositol 3 kinase (PI3-K) activity in anti-phosphotyrosine immunoprecipitates from cells expressing beta 763 as well as beta c, whereas it was only marginally increased from cells expressing beta 517 or beta 626. Thus, there are at least two distinct regions within the cytoplasmic domain of beta c that are responsible for different signals, i.e. a membrane proximal region of approximately 60 amino acid residues upstream of Glu517 is essential for induction of c-myc and pim-1, and a distal region of approximately 140 amino acid residues (between Leu626 and Ser763) is required for activation of Ras, Raf-1, MAP kinase and p70 S6 kinase, as well as induction of c-fos and c-jun. PMID- 8223434 TI - Tryptophan W207 in transducin T alpha is the fluorescence sensor of the G protein activation switch and is involved in the effector binding. AB - We have produced a recombinant transducin alpha subunit (rT alpha) in sf9 cells, using a baculovirus system. Deletion of the myristoylation site near the N terminal increased the solubility and allowed the purification of rT alpha. When reconstituted with excess T beta gamma on retinal membrane, rT alpha displayed functional characteristics of wild-type T alpha vis a vis its coupled receptor, rhodopsin and its effector, cGMP phosphodiesterase (PDE). We further mutated a tryptophan, W207, which is conserved in all G proteins and is suspected to elicit the fluorescence change correlated to their activation upon GDP/GTP exchange or aluminofluoride (AlFx) binding. [W207F]T alpha mutant displayed high affinity receptor binding and underwent a conformational switch upon receptor-catalysed GTP gamma S binding or upon AlFx binding, but this did not elicit any fluorescence change. Thus W207 is the only fluorescence sensor of the switch. Upon the switch the mutant remained unable to activate the PDE. To characterize better its effector-activating interaction we measured the affinity of [W207F]T alpha GDP-AlFx for PDE gamma, the effector subunit that binds most tightly to T alpha. [W207F]T alpha still bound in an activation-dependent way to PDE gamma, but with a 100-fold lower affinity than rT alpha. This suggests that W207 contributes to the G protein effector binding. PMID- 8223435 TI - cAMP antagonizes p21ras-directed activation of extracellular signal-regulated kinase 2 and phosphorylation of mSos nucleotide exchange factor. AB - In fibroblasts, stimulation of receptor tyrosine kinases results in the activation of the extracellular signal-regulated kinase 2 (ERK2). The major signalling pathway employed by these receptors involves the activation of p21ras and raf-1 kinase. Here we show that in NIH3T3 and rat-1 fibroblasts, elevation of the intracellular cAMP level results in the inhibition of ERK2 activation induced by PDGF, EGF and insulin treatment. Analysis of various signalling intermediates shows that cAMP interferes at a site downstream of p21ras, but upstream of raf-1 kinase. Inhibition by cAMP depends on both the cAMP concentration and the absolute amount of p21ras molecules bound to GTP, suggesting a mechanism of competitive inhibition. Also TPA-induced, p21ras-independent, activation of raf-1 kinase and ERK2 is inhibited by cAMP. We have used the inhibitory effect of cAMP to investigate whether phosphorylation of mSos, a p21ras nucleotide exchange factor, is dependent on the activity of the raf-1 kinase/ERK2 pathway. We found that phosphorylation of mSos, as monitored by a mobility shift, is delayed with respect to p21ras and ERK2 activation and is inhibited by cAMP in a similar cell type- and concentration-dependent manner as the inactivation of ERK2. These results provide evidence for a model of p21ras-directed signalling towards ERK2 that feeds back on mSos by regulating its phosphorylation status and that can be negatively modulated by protein kinase A and positively modulated by protein kinase C action. PMID- 8223436 TI - NF-kappa B activates the HIV promoter in neurons. AB - Human immunodeficiency virus (HIV) infection of the brain leads to massive neuronal damage, resulting in the AIDS (acquired immunodeficiency syndrome) dementia complex (ADC). A recent study using transgenic mice indicates that neurons possess transcription factors capable of activating the HIV promoter. To identify these, we transfected two types of primary cultures of rat neurons with HIV promoter-reporter gene constructs. The two kappa B regulatory sites in the HIV long terminal repeat (LTR) are shown to be essential for strong promoter activity. Two proteins present in neurons, BETA and an NF-kappa B-like protein, can bind the kappa B sites. These proteins are shown to belong to distinct families of transcription factors. Mutation analysis and transfection of a dominant negative NF-kappa B mutant, indicate that the neuronal NF-kappa B-like activity mediates HIV promoter activation. cDNA cloning, biochemical and immunological analyses indicate that neuronal NF-kappa B is similar to NF-kappa B of other tissues. Transfections of primary neuron cultures with an HIV promoter beta-galactosidase construct show that within these cultures, neurons are indeed the cells that highly activate the HIV promoter. Thus, analogous to the situation in T-lymphocytes and macrophages, NF-kappa B is an activator of HIV transcription in neurons. PMID- 8223437 TI - Functional and biochemical interaction of the HTLV-I Tax1 transactivator with TBP. AB - The human T-cell leukemia virus type I (HTLV-I) codes for the potent transcriptional activator, Tax1, which induces the enhancer activity of various enhancer elements. In the case of the 21 bp enhancer of the HTLV-I provirus, this induction is correlated with the association of Tax1 with this DNA element via a specific cellular factor. That the indirect association of Tax1 with DNA can lead to transcriptional activation has also been supported by the study of chimeric GAL4-Tax1 proteins. The GAL4-Tax1 stimulatory effect exhibits a strong self squelching. In order to determine whether Tax1 interacts directly with the general transcription factors or via intermediary molecules, we have analyzed how overexpression of the TATA binding protein (TBP) and TFIIB protein affects the squelching curve of GAL4-Tax1. The data presented here show that overexpression of TBP strongly increases the stimulatory effect of GAL4-Tax1, causes a displacement of the maximum of the squelching curve and partially alleviates the squelching. Under similar conditions TFIIB exhibited little effect. From these results we conclude that Tax1 can increase the recruitment of TBP by directly interacting with this protein. Biochemical experiments with purified proteins produced in bacteria confirmed that Tax1 can interact with TBP but not with TFIIB. Tax1 interacts with the conserved C-terminal part of TBP. Analysis of the ability of different mutants of Tax1 fused to the GAL4 DNA binding domain to activate transcription and to associate with TBP, showed that these activities are correlated. However, since one transcriptionally inactive mutant was able to interact efficiently with TBP in vitro, it would appear that an event other than the Tax1-TBP contact also intervenes in the activation of transcription by Tax1. PMID- 8223438 TI - A human homologue of Saccharomyces cerevisiae SNF2/SWI2 and Drosophila brm genes potentiates transcriptional activation by the glucocorticoid receptor. AB - Several of the SNF and SWI genes of Saccharomyces cerevisiae code for proteins believed to assist transcriptional activators by relieving nucleosome repression. One of these proteins, SNF2/SWI2, has a homologue in Drosophila, a regulator of homeotic genes known as brahma or brm. In this report, we show that a counterpart of SNF2/SWI2 also exists in mice and humans. The human protein, designated hbrm, is a 180 kDa nuclear factor that can function as a transcriptional activator when fused to a heterologous DNA binding domain. The mouse homologue of hbrm is expressed in all mouse organs tested while hbrm was detected in some but not all investigated human cell lines. In cells failing to express the endogenous gene, transfected hbrm cooperates with the glucocorticoid receptor (GR) in transcriptional activation. However, hbrm had no effect on the activity of several other transcription factors, including the homeoprotein HNF-1. The co operation between hbrm and GR required the DNA binding domain of GR and two separated regions of the hbrm protein, including a domain with homology to known helicases. PMID- 8223439 TI - Editing of the chloroplast rpoB transcript is independent of chloroplast translation and shows different patterns in barley and maize. AB - Sequence analysis of amplified cDNAs derived from the maize chloroplast rpoB transcript which encodes the beta subunit of a chloroplast specific, DNA dependent RNA polymerase reveals four C-to-U editing sites clustered within 150 nucleotides of the 5' terminal region of the rpoB message. These newly identified editing sites confirm the bias of chloroplast editing for certain codon transitions and for second codon positions which both appear suggestive for an involvement of the translational apparatus in the editing process. This supposition prompted us to investigate editing of the rpoB transcript from ribosome deficient, and hence protein synthesis deficient, plastids of the barley mutant albostrians. In this mutant editing is, however, not impaired at any of the editing sites functional in the barley wild type rpoB transcript. This demonstrates that chloroplast editing is neither linked to nor dependent on the chloroplast translational apparatus. As a further consequence any peptide components required for chloroplast editing must be encoded in the nuclear genome. In spite of strong sequence conservation only three of the four editing sites identified in the maize rpoB transcript are functional in barley. This indicates that sequences surrounding an editing site alone are not sufficient as determinants for the editing process in chloroplasts, but that trans-acting templates carrying the editing information for each individual site may also be required. PMID- 8223440 TI - Depletion of functional ribosomal RNA operons in Escherichia coli causes increased expression of the remaining intact copies. AB - The synthesis of ribosomal RNA is a complex and highly regulated process. To study this process, we have used deletion-insertions to disrupt sequentially from one to four of the seven rRNA (rrn) operons on the Escherichia coli genome. Inactivation of four rrn operons caused a 2.3-fold increase in the expression of a chloramphenicol acetyl transferase reporter gene fused to the tandem promoters of rrnA and a similar increase in the expression of the trp tRNA gene at the end of rrnC. This reflected enhanced expression of the remaining operons to compensate for having only three intact copies. The elevated expression was caused by an increase in both transcription initiation and RNA polymerase elongation rates specifically on rrn operons and occurred in the absence of changes in the intracellular concentration of ppGpp, suggesting that ppGpp is not involved in the regulation of this phenomenon. We discuss these results in relation to the ribosome feedback inhibition model described by Nomura and coworkers. PMID- 8223441 TI - Functional synergy between DP-1 and E2F-1 in the cell cycle-regulating transcription factor DRTF1/E2F. AB - It is widely believed that the cellular transcription factor DRTF1/E2F integrates cell cycle events with the transcription apparatus because during cell cycle progression in mammalian cells it interacts with molecules that are important regulators of cellular proliferation, such as the retinoblastoma tumour suppressor gene product (pRb), p107, cyclins and cyclin-dependent kinases. Thus, pRb, which negatively regulates early cell cycle progression and is frequently mutated in tumour cells, and the Rb-related protein p107, bind to and repress the transcriptional activity of DRTF1/E2F. Viral oncoproteins, such as adenovirus E1a and SV40 large T antigen, overcome such repression by sequestering pRb and p107 and in so doing are likely to activate genes regulated by DRTF1/E2F, such as cdc2, c-myc and DHFR. Two sequence-specific DNA binding proteins, E2F-1 and DP-1, which bind to the E2F site, contain a small region of similarity. The functional relationship between them has, however, been unclear. We report here that DP-1 and E2F-1 exist in a DNA binding complex in vivo and that they bind efficiently and preferentially as a heterodimer to the E2F site. Moreover, studies in yeast and Drosophila cells indicate that DP-1 and E2F-1 interact synergistically in E2F site-dependent transcriptional activation. PMID- 8223442 TI - The activity of S.pombe DSC-1-like factor is cell cycle regulated and dependent on the activity of p34cdc2. AB - In the eukaryotic cell cycle, there are major control points in late G2 to determine the timing of the initiation of mitosis, and in late G1, regulating entry into S phase. In yeasts, this latter control is called start. Traverse of the start control and progression to S phase is accompanied by an increase in the expression of some of the genes whose products are required for DNA synthesis. In Saccharomyces cerevisiae, the coordinate expression of these genes in late G1 is dependent on a cis-acting sequence element called the MluI cell cycle box (MCB). A transcription factor called DSC-1 binds these elements and mediates cell cycle regulated transcription, though it is unclear whether this is by cell cycle dependent changes in its activity. A DSC-1-like factor has also been identified in the fission yeast S.pombe. This is composed of at least the products of the cdc10 and sct1/res1 genes, and binds to the promoters of genes whose expression increases prior to S phase. We demonstrate that p85cdc10 is a nuclear protein and that the activity of the S.pombe DSC-1 factor varies through the cell cycle; it is high in cells that have passed start, decreases at the time of anaphase, remains low during the pre-start phase of G1 and increases at the time of the next S phase. We also show that the reactivation in late G1 is dependent on the G1 form of p34cdc2. PMID- 8223443 TI - TCR beta and TCR alpha gene enhancers confer tissue- and stage-specificity on V(D)J recombination events. AB - We describe transgenic mice carrying germline variable gene segments associated with either the T cell receptor (TCR) beta or alpha gene enhancers (E beta or E alpha). Transgenic constructs underwent high rates of site-specific rearrangements predominantly in T cells from independent mice. Rearrangements of the E beta-containing transgenes began at different stages of T cell differentiation in embryonic and adult thymus than did the E alpha-containing ones, with a pattern superimposable upon the patterns of TCR beta or TCR alpha gene expression, respectively. We demonstrate that sequences within the TCR beta and TCR alpha gene enhancers confer tissue- and stage-specificity upon the V(D)J recombination events affecting adjacent gene segments. The patterns of transgene expression also gave information on developmental events and lineage relationships (gamma delta versus alpha beta) during T cell development. PMID- 8223444 TI - T cell development in mice lacking the CD3-zeta/eta gene. AB - The CD3-zeta and CD3-eta polypeptides are two of the components of the T cell antigen receptor (TCR) which contribute to its efficient cell surface expression and account for part of its transducing capability. CD3-zeta and CD3-eta result from the alternative splicing of a single gene designated CD3-zeta/eta. To evaluate the role of these subunits during T cell development, we have produced mice with a disrupted CD3-zeta/eta gene. The analysis of thymocyte populations from the CD3-zeta/eta-/- homozygous mutant mice revealed that they have a profound reduction in the surface levels of TCR complexes and that the products of the CD3-zeta/eta gene appear to be needed for the efficient generation and/or survival of CD4+CD8+ thymocytes. Despite the almost total absence of mature single positive thymocytes, the lymph nodes from zeta/eta-/- mice were found to contain unusual CD4+CD8- and CD4-CD8+ single positive cells which were CD3-. In contrast to the situation observed in the thymus, the thymus-independent gut intraepithelial lymphocytes present in zeta/eta-/- mice do express TCR complexes on their surface and these are associated with Fc epsilon RI gamma homodimers. These results establish an essential role for the CD3-zeta/eta gene products during intrathymic T cell differentiation and further emphasize the difference between conventional T cells and thymus-independent gut intraepithelial lymphocytes. PMID- 8223445 TI - Developmental and functional impairment of T cells in mice lacking CD3 zeta chains. AB - CD3 zeta is a component of the T cell antigen receptor (TCR) complex and is important for signal transduction. We have established mice selectively lacking CD3 zeta but able to express CD3 eta, a polypeptide produced from the same locus through alternative splicing, using the method of gene targeting in embryonic stem cells. In homozygous mutant mice, the numbers of thymocytes and peripheral T cells were greatly reduced and the expression levels of TCR on these cells were 5 fold lower than those on wild-type cells. By contrast, TCR gamma delta+ intestinal intraepithelial lymphocytes were not obviously affected by the mutation. T cells from homozygous mutants exhibited an impaired proliferative response. These results imply that CD3 zeta has a critical role in the development and signal transduction of T cells in vivo. PMID- 8223446 TI - Raf-1 is required for T cell IL2 production. AB - Engagement of the T cell receptor/CD3 complex activates the serine/threonine kinase, Raf-1, but the physiologic consequences of its activation have not been determined. The effects of Raf-1 on interleukin 2 (IL2) production in T cells were examined using activated and inhibitory forms of Raf-1. A truncated active form of Raf-1 was expressed constitutively from the metallothionein promoter in a malignant T cell line, Jurkat. Treatment of the cells with zinc and cadmium greatly increased active Raf-1 expression. This increase in Raf-1 expression allowed antibodies to CD3 and to CD28 to stimulate IL2 production in the absence of phorbol myristate acetate (PMA) and enhanced IL2 production stimulated by these antibodies in the presence of PMA. The action of active Raf-1 was to increase IL2 gene transcription as it enhanced transcription of a reporter gene linked to IL2 promoter. Finally, the dominant negative form of Raf-1 inhibited transcription directed by the IL2 promoter that was induced by the mitogen phytohemagglutinin (PHA) and PMA. We conclude that Raf-1 activity is necessary for IL2 gene transcription and secretion. These data indicate a role for Raf-1 in the immune response. PMID- 8223447 TI - Expression of conformationally constrained adhesion peptide in an antibody CDR loop and inhibition of natural killer cell cytotoxic activity by an antibody antigenized with the RGD motif. AB - We report that an antibody engineered to express three Arg-Gly-Asp (RGD) repeats in the third complementarity-determining region of the heavy chain (antigenized antibody) efficiently inhibits the lysis of human erythroleukemia K-562 cells by natural killer (NK) cells. Synthetic peptides containing RGD did not inhibit. Inhibition was specific for the (RGD)3-containing loop and required simultaneous occupancy of the Fc receptor (CD16) on effector cells. The antigenized antibody inhibited other forms of cytotoxicity mediated by NK cells but not cytotoxicity mediated by major histocompatibility complex-restricted cytotoxic T lymphocytes (CTL). A three-dimensional model of the engineered antibody loop shows the structure and physicochemical characteristics probably required for the ligand activity. The results indicate that an RGD motif is involved in the productive interaction between NK and target cells. Moreover, they show that peptide expression in the hypervariable loops of an antibody molecule is an efficient procedure for stabilizing oligopeptides within a limited spectrum of tertiary structures. This is a new approach towards imparting ligand properties to antibody molecules and can be used to study the biological function and specificity of short peptide motifs, including those involved in cell adhesion. PMID- 8223448 TI - A novel type of class I gene organization in vertebrates: a large family of non MHC-linked class I genes is expressed at the RNA level in the amphibian Xenopus. AB - A Xenopus class I cDNA clone, isolated from a cDNA expression library using antisera, is a member of a large family of non-classical class I genes (class Ib) composed of at least nine subfamilies, all of which are expressed at the RNA level. The subfamilies are well conserved in their immunoglobulin-like alpha 3 domains, but their peptide-binding regions (PBRs) and cytoplasmic domains are very divergent. In contrast to the great allelic diversity found in the PBR of classical class I genes, the alleles of one of the Xenopus non-classical subfamilies are extremely well conserved in all regions. Several of the invariant amino acids essential for the anchoring of peptides in the classical class I groove are not conserved in some subfamilies, but the class Ib genes are nevertheless more closely related in the PBR to classical and non-classical genes linked to the MHC in mammals and birds than to any other described class I genes like CD1 and the neonatal rat intestinal Fc receptor. Comparison with the Xenopus MHC-linked class Ia protein indicate that amino acids presumed to interact with beta 2-microglobulin are identical or conservatively changed in the two major class I families. Genomic analyses of Xenopus species suggest that the classical and non-classical families diverged from a common ancestor before the emergence of the genus Xenopus over 100 million years ago; all of the non-classical genes appear to be linked on a chromosome distinct from the one harboring the MHC. We hypothesize that this class Ib gene family is under very different selection pressures from the classical MHC genes, and that each subfamily may have evolved for a particular function. PMID- 8223449 TI - Cell fusion-induced quick change in replication time of the inactive mouse X chromosome: an implication for the maintenance mechanism of late replication. AB - It is unknown how and why the genetically inactivated mammalian X chromosome replicates late in S phase. There are also occasional inactive X chromosomes characterized by an opposite behavior replicating early in S phase. Two clonal cell lines, MTLB3 and MTLH8, isolated from a cultured murine T-cell lymphoma have an allocyclic X chromosome of the latter type. This precociously replicating X chromosome was judged to be genetically inactive as the late replicating one. Immediately after fusion with another cell line, the precociously replicating X chromosome from these cells starts to replicate late in S phase. This finding seems to suggest that late replication characterizing the inactive X chromosome is actively maintained by a trans-acting factor in female somatic cells, and that its lack entails a switch from late replication to precocious replication. It remains unknown whether this presumptive factor also modifies the autosomal replication pattern. PMID- 8223450 TI - DNA curvature influences the internal motions of supercoiled DNA. AB - We present evidence that short curved DNA segments can act as mediators for the ordering of large domains in superhelical DNA. Using a non-invasive solution method (dynamic light scattering), we investigated the effect of permanently curved inserts on the solution structure and on the internal motions of superhelical plasmid DNA. We find that the dynamics of superhelical DNA are strongly influenced by sequence- or protein-induced bending: in superhelical plasmids containing curved inserts the amplitude of the internal motion is lower than that of non-curved controls. Furthermore, the relative arrangement of curved sequences in the plasmids can influence the overall shape of the superhelical DNA. On linearized forms of the plasmids, these effects are not observed. PMID- 8223451 TI - The alpha-helical rod domain of human lamins A and C contains a chromatin binding site. AB - We examined regions of human lamins A and C involved in binding to surfaces of mitotic chromosomes. An Escherichia coli expression system was used to produce full-length lamin A and lamin C, and truncated lamins retaining the central alpha helical rod domain (residues 34-388) but lacking various amounts of the amino terminal 'head' and carboxy-terminal 'tail' domains. We found that lamin A, lamin C and lamin fragments lacking the head domain and tail sequences distal to residue 431 efficiently assembled into paracrystals and strongly associated with mitotic chromosomes. Furthermore, the lamin rod domain also associated with chromosomes, although efficient chromosome coating required the pH 5-6 conditions needed to assemble the rod into higher order structures. Biochemical assays showed that chromosomes substantially reduced the critical concentration for assembly of lamin polypeptides into pelletable structures. Association of the lamin rod with chromosomes was abolished by pretrypsinization of chromosomes, and was not seen for vimentin (which possesses a similar rod domain). These data demonstrate that the alpha-helical rod of lamins A and C contains a specific chromosome binding site. Hence, the central rod domain of intermediate filament proteins can be involved in interactions with other cellular structures as well as in filament assembly. PMID- 8223452 TI - dUTP pyrophosphatase is an essential enzyme in Saccharomyces cerevisiae. AB - dUTP pyrophosphatase (dUTPase; EC 3.6.1.23) catalyses the hydrolysis of dUTP to dUMP and PPi and thereby prevents the incorporation of uracil into DNA during replication. Although it is widely believed that dUTPase is essential for cell viability because of this role, direct evidence supporting this assumption has not been presented for any eukaryotic system. We have analysed the role of dUTPase (DUT1) in the life cycle of yeast. Using gene disruption and tetrad analysis, we find that DUT1 is necessary for the viability of S. cerevisiae; however, under certain conditions dut1 null mutants survive if supplied with exogenous thymidylate (dTMP). Analyses with isogenic uracil-DNA-glycosylase (UNG1) deficient or proficient strains indicate that in the absence of dUTPase, cell death results from the incorporation of uracil into DNA and the attempted repair of this damage by UNG1-mediated excision repair. However, in dut1 ung1 double mutants, starvation for dTMP causes dividing cells to arrest and die in all phases of the cell cycle. This latter effect suggests that the extensive stable substitution of uracil for thymine in DNA leads to a general failure in macromolecular synthesis. These results are in general agreement with previous models in thymine-less death that implicate dUTP metabolism. They also suggest an alternative approach for chemotherapeutic drug design. PMID- 8223453 TI - A defective human foamy provirus generated by pregenome splicing. AB - Foamy viruses are a group of retroviruses of complex structure which were thought to be non-pathogenic. The recent demonstration of neurological diseases in mice transgenic for human foamy virus (HFV) and the high prevalence of HFV sequences in Graves' disease question this idea. By PCR, we have detected HFV sequences with a non-random deletion in the bel1 transactivator gene in other autoimmune conditions. Sequence analysis revealed that this deleted area corresponds to the excision of a known intron in bet, one of HFV's regulatory genes. The same phenomenon was observed in both acute and chronic infections, in vitro or in vivo, although the deleted forms were distinctly more abundant in chronic states. The viral DNA containing the bel1 deletion is apparently part of an otherwise complete genome, strongly suggesting that this provirus derives from the reverse transcription of a spliced pregenomic RNA. Bel1-spliced provirus was shown to be defective when transfected into permissive cells. However, co-expression with the Bel1 transactivator led to functional trans-complementation and formation of viral particles. Splicing of the genome may be an important factor in HFV biology: genomes with the deletion may either interfere with wild-type virus expression or alter host cell functions through background expression of viral regulatory proteins. PMID- 8223454 TI - Identification of the initiation sequence for viral-strand DNA synthesis of wheat dwarf virus. AB - The intergenic region of the circular single-stranded DNA genome of geminiviruses contains a sequence potentially able to fold into a stem-loop structure. This sequence has been reported to be involved in viral replication by serving as the origin for rolling-circle replication. However, in wheat dwarf virus (WDV) a deletion of 128 bp, removing this sequence, surprisingly does not prevent de novo viral DNA synthesis, but instead abrogates the processing of replicative intermediates into monomeric genomes. This deletion mutant permitted us to study the initiation of viral-strand DNA synthesis independently from its termination and also to identify the sequence within which rolling-circle DNA replication of WDV begins. We have mapped the initiation site of replication to a pentanucleotide, TACCC, a sequence that occurs twice in the large intergenic region of WDV: it is found in the right half of the stem-loop sequence and again 170 bases upstream where it is part of a 15 nucleotide sequence highly homologous to the right half of the stem-loop sequence. Here we show that viral-strand DNA synthesis efficiently initiates at both sequences. PMID- 8223455 TI - RNA-mediated specificity of DNA packaging into hybrid lambda/phi 29 proheads. AB - A small RNA (pRNA, 174 nt) is known to be essential for DNA packaging in bacteriophage phi 29. However, in an in vitro DNA packaging system based on hybrid lambda/phi 29 proheads (made up of head proteins from phage lambda and connectors from phage phi 29), the specificity of DNA packaging is lost, and different RNA molecules fulfil the requirements for DNA packaging, albeit with less efficiency than phi 29 pRNA. Competition assays with RNAs from different sources have shown that phi 29 connectors bind preferentially pRNA. An increase in the efficiency of phi 29 DNA packaging into hybrid proheads induced by phi 29 pRNA is observed because, when phi 29 pRNA is incubated with hybrid proheads, phi 29 DNA is packaged more efficiently than other DNAs of similar length. Furthermore, when hybrid proheads carrying phi 29 pRNA are incubated with a mixture of DNAs from different sources, phi 29 DNA is selectively packaged, thus indicating that phi 29 pRNA determines the specificity of DNA packaging. PMID- 8223456 TI - Mesoderm-inducing factors in early vertebrate development. PMID- 8223457 TI - Secretion and mesoderm-inducing activity of the TGF-beta-related domain of Xenopus Vg1. AB - Vg1 is a maternal mRNA localized to the vegetal hemisphere of Xenopus embryos during blastula stages, a region responsible for the induction of mesoderm in the adjacent marginal zone. Its homology to the transforming growth factor-beta family, which includes several proteins with mesoderm-inducing activity, suggests a role for Vg1 as an endogenous mesoderm-inducing factor. However, expression of Vg1 protein in the animal hemisphere, following injection of synthetic mRNA, has no effect on development, and isolated animal caps are not mesodermalized. It is shown that Vg1 protein fails to form dimers and is not processed to release the putative bioactive domain. Furthermore it is shown that the N-terminal signal peptide of Vg1 is not cleaved following translocation into the ER, which may explain the failure of this protein to dimerize. To explore the role of Vg1 in amphibian development, a fusion protein has been made of the preproregion of Xenopus bone morphogenetic protein-4 and the putative bioactive C-terminal domain of Vg1. This fusion protein forms dimers and the C-terminal domain of Vg1 is secreted. Injection of this construct into Xenopus embryos induces the formation of a second dorsal axis and isolated animal caps are mesodermalized. The results are consistent with a role for Vg1 in mesoderm induction during Xenopus development. PMID- 8223458 TI - Combinatorial generation of variable fusion proteins in the Ewing family of tumours. AB - Balanced translocations involving band q12 of human chromosome 22 are the most frequent recurrent translocations observed in human solid tumours. It has been shown recently that this region encodes EWS, a protein with an RNA binding homologous domain. In Ewing's sarcoma and malignant melanoma of soft parts, translocations of band 22q12 to chromosome 11 and 12 result in the fusion of EWS with the transcription factors FLI-1 and ATF1, respectively. The present analysis of 89 Ewing's sarcomas and related tumours show that in addition to the expected EWS-FLI-1 fusion, the EWS gene can be fused to ERG, a transcription factor closely related to FLI-1 but located on chromosome 21. The position of the chromosome translocation breakpoints are shown to be restricted to introns 7-10 of the EWS gene and widely dispersed within introns 3-9 of the Ets-related genes. This heterogeneity generates a variety of chimeric proteins that can be detected by immuno-precipitation. On rare occasions, they may be associated with a truncated EWS protein arising from alternate splicing. All 13 different fusion proteins that were evidenced contained the N-terminal domain of EWS and the Ets domain of FLI-1 or ERG suggesting that oncogenic conversion is achieved by the linking of the two domains with no marked constraint on the connecting peptide. PMID- 8223459 TI - Hybrid cell extinction and re-expression of Oct-3 function correlates with differentiation potential. AB - The Oct-3 gene is expressed in highly undifferentiated cells and is implicated in mammalian early embryogenesis. We have generated a series of hybrid cells between pluripotent embryonal carcinoma cells (Oct-3+) and fibroblasts (Oct-3-), and have studied the regulation and function of Oct-3. Upon fusion, the hybrid cells differentiated to nestin+/Brn-2+ cells resembling neuroepithelial stem cells. Expression of Oct-3 was extinguished at the transcriptional level in all the hybrid cells examined. The Oct-3 modulating activity required for the Oct-3 mediated enhancer activation was also extinguished. When the Oct-3 transactivating function was introduced into the hybrid cells, they transformed into morphologically distinct nestin-/Brn-2- cells ('revertants'). When the 'revertant' cells subsequently lost Oct-3 expression, they differentiated back to nestin+/Brn-2+ cells. The close correlation between the phenotypic changes and the gain/loss of Oct-3 function indicates that Oct-3 can induce dedifferentiation of the neural cells. PMID- 8223460 TI - Role of the gooseberry gene in Drosophila embryos: maintenance of wingless expression by a wingless--gooseberry autoregulatory loop. AB - During Drosophila embryogenesis, segment polarity genes, such as engrailed (en), wingless (wg) and gooseberry (gsb) show complex interactions that provide positional information along the antero-posterior axis within each segment. Little is known about the specific role of each of these genes in this pattern determining process. Here we demonstrate that the main function of gsb, which encodes a transcription factor containing a paired-domain and a prd-type homeodomain, is the maintenance of wg expression by a wg-gsb autoregulatory loop after 6 h of development. The function of wg, the homologue of the murine Wnt-1 gene, is to specify the denticle pattern by repressing a default state of ubiquitous denticle formation in the ventral epidermis. This repression of denticles by the wg signal is different from the wingless signalling pathways that activate gsb or en. Mutual activations involving gsb, wg and en show temporal asymmetries that lead to their different mutant phenotypes. A general model is proposed for the generation of morphogenetic fields by self-propagating autoregulatory loops. PMID- 8223461 TI - Chromosomal replication initiates and terminates at random sequences but at regular intervals in the ribosomal DNA of Xenopus early embryos. AB - We have analysed the replication of the chromosomal ribosomal DNA (rDNA) cluster in Xenopus embryos before the midblastula transition. Two-dimensional gel analysis showed that replication forks are associated with the nuclear matrix, as in differentiated cells, and gave no evidence for single-stranded replication intermediates (RIs). Bubbles, simple forks and double Ys were found in each restriction fragment analysed, showing that replication initiates and terminates without detectable sequence specificity. Quantification of the results and mathematical analysis showed that the average rDNA replicon replicates in 7.5 min and is 9-12 kbp in length. This time is close to the total S phase duration, and this replicon size is close to the maximum length of DNA which can be replicated from a single origin within this short S phase. We therefore infer that (i) most rDNA origins must be synchronously activated soon in S phase and (ii) origins must be evenly spaced, in order that no stretch of chromosomal DNA is left unreplicated at the end of S phase. Since origins are not specific sequences, it is suggested that this spatially and temporally concerted pattern of initiation matches some periodic chromatin folding, which itself need not rely on DNA sequence. PMID- 8223462 TI - A DNA unwinding element and an ARS consensus comprise a replication origin within a yeast chromosome. AB - We have defined a replication origin, ORI305, within chromosome III of Saccharomyces cerevisiae by means of mutational analysis. cis-acting elements required for origin activity in the chromosome, as assayed by two-dimensional gel electrophoresis of replication intermediates, are the same as those required for the function of an autonomously replicating sequence, ARS305, in a plasmid. Essential elements include (i) an 11 bp sequence that is a near match to the ARS consensus and (ii) a broad sequence directly 3' to the consensus near match. Origin function is inactivated by point mutations in the essential near match sequence, suggesting that the sequence contributes to specifying the origin in the chromosome. Other consensus near matches with different sequences are present but are not required. The essential 3'-flanking sequence exhibits DNA helical instability and is sensitive to deletion mutations that stabilize the DNA helix. The wild-type 3'-flanking sequence can be functionally substituted by dissimilar sequences that also exhibit helical instability. The requirement for DNA helical instability indicates that the essential 3'-flanking sequence serves as a DNA unwinding element in the chromosome. PMID- 8223463 TI - Disruption of the nucleosomes at the replication fork. AB - The fate of parental nucleosomes during chromatin replication was studied in vitro using in vitro assembled chromatin containing the whole SV40 genome as well as salt-treated and native SV40 minichromosomes. In vitro assembled minichromosomes were able to replicate efficiently in vitro, when the DNA was preincubated with T-antigen, a cytosolic S100 extract and three deoxynucleoside triphosphates prior to chromatin assembly, indicating that the origin has to be free of nucleosomes for replication initiation. The chromatin structure of the newly synthesized daughter strands in replicating molecules was analysed by psoralen cross-linking of the DNA and by micrococcal nuclease digestion. A 5- and 10-fold excess of protein-free competitor DNA present during minichromosome replication traps the segregating histones. In opposition to published data this suggests that the parental histones remain only loosely or not attached to the DNA in the region of the replication fork. Replication in the putative absence of free histones shows that a subnucleosomal particle is randomly assembled on the daughter strands. The data are compatible with the formation of a H3/H4 tetramer complex under these conditions, supporting the notion that under physiological conditions nucleosome core assembly on the newly synthesized daughter strands occurs by the binding of H2A/H2B dimers to a H3/H4 tetramer complex. PMID- 8223464 TI - Open-complex formation by the host initiator, DnaA, at the origin of P1 plasmid replication. AB - Replication of P1 plasmid requires both the plasmid-specific initiator, RepA, and the host initiator, DnaA. Here we show that DnaA can make the P1 origin reactive to the single-strand specific reagents KMnO4 and mung bean nuclease. Addition of RepA further increased the KMnO4 reactivity of the origin, although RepA alone did not influence the reaction. The increased reactivity implies that the two initiators interact in some way to alter the origin conformation. The KMnO4 reactivity was restricted to one strand of the origin. We suggest that the roles of DnaA in P1 plasmid and bacterial replication are similar: origin opening and loading of the DnaB helicase. The strand-bias in chemical reactivity at the P1 origin most likely indicates that only one of the strands is used for the loading of DnaB, a scenario consistent with the unidirectional replication of the plasmid. PMID- 8223465 TI - The role of the 70 kDa subunit of human DNA polymerase alpha in DNA replication. AB - DNA polymerase alpha is the only enzyme in eukaryotic cells capable of starting DNA chains de novo and is required for the initiation of SV40 DNA replication in vitro. We have cloned the 70 kDa subunit of human DNA polymerase alpha (hereafter referred to as the B subunit) and expressed it as a fusion protein in bacteria. The purified fusion protein forms a stable complex with SV40 T antigen, both in solution and when T antigen is bound to the SV40 origin of DNA replication. Analysis of mutant forms of the B subunit indicates that the N-terminal 240 amino acids are sufficient to mediate complex formation. The B subunit fusion protein promotes formation of a complex containing T antigen and the catalytic subunit (subunit A) of DNA polymerase alpha, suggesting that it serves to tether the two proteins. These physical interactions are functionally significant, since the ability of T antigen to stimulate the activity of the catalytic subunit of DNA polymerase alpha is highly dependent upon the B subunit. We suggest that the interactions mediated by the B subunit play an important role in SV40 DNA replication by promoting DNA chain initiation at the origin and/or facilitating the subsequent priming and synthesis of DNA chains on the lagging strand template. The protein may play similar roles in cellular DNA replication. PMID- 8223466 TI - Synaptic intermediates in bacteriophage lambda site-specific recombination: integrase can align pairs of attachment sites. AB - Bacteriophage lambda uses site-specific recombination to move its DNA into and out of the Escherichia coli genome. The recombination event is mediated by the recombinase integrase (Int) together with several accessory proteins through short specific DNA sequences known as attachment sites. A gel mobility shift assay has been used to show that, in the absence of accessory proteins, Int can align and hold together two DNA molecules, each with an attachment site, to form stable non-covalent 'bimolecular complexes'. Each attachment site must have both core and arm binding sites for Int to participate in a bimolecular complex. These stable structures can be formed between pairs of attL and attP attachment sites, but cannot include attB or attR sites; they are inhibited by integration host factor (IHF) protein. The bimolecular complexes are shown to represent a synaptic intermediate in the reaction in which Int protein promotes the IHF-independent recombination of two attL sites. These complexes should enable a detailed analysis of synapsis for this pathway. PMID- 8223467 TI - Complementation of bacteriophage lambda integrase mutants: evidence for an intersubunit active site. AB - Site-specific recombination of bacteriophage lambda starts with the formation of higher-order protein--DNA complexes, called 'intasomes', and is followed by a series of steps, including the initial DNA cleavage, top-strand exchange, branch migration and bottom-strand exchange, to produce recombinant products. One of the intasomes formed during excisive recombination (the attL complex) is composed of the phage-encoded integrase (Int), integration host factor (IHF) and one of the recombination substrates, attL DNA. Int is the catalytic recombinase and has two different DNA binding domains. When IHF is present, Int binds to two types of sites in attL DNA, the three arm-type sites (P'123) and the core-type sites (B and C') where the reciprocal strand exchange takes place. The Tyr342 residue of Int serves as a nucleophile during strand cleavage and covalently attaches to the DNA through a phosphotyrosyl bond. In vitro complementation assays have been performed for strand cleavage using attL suicide substrates and mutant proteins containing amino acid substitutions at residues conserved in the integrase family of recombinases. We demonstrate that at least two Int monomers are required to form the catalytically-competent species that performs cleavage at the B site. It is likely that the active site is formed by two Int monomers. PMID- 8223468 TI - Macroevolution by transposition: drastic modification of DNA recognition by a type I restriction enzyme following Tn5 transposition. AB - We have characterized a novel mutant of EcoDXXI, a type IC DNA restriction and modification (R-M) system, in which the specificity has been altered due to a Tn5 insertion into the middle of hsdS, the gene which encodes the polypeptide that confers DNA sequence specificity to both the restriction and the modification reactions. Like other type I enzymes, the wild type EcoDXXI recognizes a sequence composed of two asymmetrical half sites separated by a spacer region: TCA(N7)RTTC. Purification of the EcoDXXI mutant methylase and subsequent in vitro DNA methylation assays identified the mutant recognition sequence as an interrupted palindrome, TCA(N8)TGA, in which the 5' half site of the wild type site is repeated in inverse orientation. The additional base pair in the non specific spacer of the mutant recognition sequence maintains the proper spacing between the two methylatable adenine groups. Sequencing of both the wild type and mutant EcoDXXI hsdS genes showed that the Tn5 insertion occurred at nucleotide 673 of the 1221 bp gene. This effectively deletes the entire carboxyl-terminal DNA binding domain which recognizes the 3' half of the EcoDXXI binding site. The truncated hsdS gene still encodes both the amino-terminal DNA binding domain and the conserved repeated sequence that defines the length of the recognition site spacer region. We propose that the EcoDXXI mutant methylase utilizes two truncated hsdS subunits to recognize its binding site. The implications of this finding in terms of subunit interactions and the malleability of the type I R-M systems will be discussed. PMID- 8223469 TI - Developmental precise excision of Oxytricha trifallax telomere-bearing elements and formation of circles closed by a copy of the flanking target duplication. AB - The 4.1 kbp TBE1 elements of Oxytricha fallax and Oxytricha trifallax are deduced to transpose into a centrisymmetric target, CAnTG, and to duplicate the central AnT. Despite conserved C(A4C4)2 telomeric repeats at their tips, free TBE1s found during macronuclear development are not linear but 4.1 kbp circles closed on one copy of the AnT target duplication. The macronucleus-destined flanks are rejoined to regenerate the target, effecting efficient and precise somatic reversion of the germline transpositional mutation. A model is presented in which transposase catalyzes concerted precise rejoining of the flanks and cyclization of the excised element. PMID- 8223470 TI - Chromatin transitions during activation and repression of galactose-regulated genes in yeast. AB - To study the fate of nucleosomes during transcription, a yeast gene 'GAL-URARIB' was constructed which is tightly regulated by the GAL1 promoter and shows in its inactive state a series of positioned nucleosomes that are sensitive for monitoring structural changes by micrococcal nuclease. Upon transcriptional activation, nucleosome positions were lost, but a residual nucleosomal repeat with an altered repeat length and no changes in psoralen accessibility measured by a band shift assay indicated that nucleosomes were present but rearranged on the transcribed gene. When chromatin was prepared 10 or 50 min after glucose repression, nucleosomes were repositioned in a large fraction of the population by a rapid process which most likely did not depend on histone synthesis or DNA replication. However, complete regeneration of the inactive structure and repeat length was observed after one cell generation (2.5 h) suggesting that in this step some missing histones were replaced. The results are consistent with a local dissociation of nucleosomes at the site of the polymerase followed by a rapid reassembly into nucleosomes behind it. The data are further supported by analysis of the chromosomal GAL1, GAL7 and GAL10 genes. PMID- 8223471 TI - Promoter, enhancer and silencer elements regulate rearrangement of an immunoglobulin transgene. AB - The chicken Ig lambda light chain locus is composed of a single V gene closely linked (1.8 kb) to a single J-C unit in its natural configuration. In mice transgenic for this locus, the transgene becomes rearranged in B cells and to a much lesser extent in T cells. Modifications were introduced in the transgene in order to characterize elements which target the recombinase to the Ig loci. In the absence of either the promoter or the enhancer located 3' of C lambda, rearrangement of the transgene is reduced 20- to 100-fold. Moreover, rearrangement is increased 5-fold when the DNA segment between V lambda and J lambda ('Uo segment'), which is deleted during the joining process, is replaced by a neutral DNA segment of equal length. The Uo segment behaved as a strong transcriptional silencer when tested in a CAT assay in vitro. Control transgenic mice harbouring only the two 3 bp mutations that introduced restriction sites at both ends of the Uo segment to allow for its replacement were also analysed. Rearrangement was reduced 10- to 100-fold in B cells from such transgenic lines. A model is proposed whereby the sites of these two mutations would function by counteracting transiently the repressing effect of the silencer, thus giving access of the chicken light chain locus to the recombinase. PMID- 8223472 TI - DNA and redox state induced conformational changes in the DNA-binding domain of the Myb oncoprotein. AB - The DNA-binding domain of the oncoprotein Myb comprises three imperfect repeats, R1, R2 and R3. Only R2 and R3 are required for sequence-specific DNA-binding. Both are assumed to contain helix-turn-helix (HTH)-related motifs, but multidimensional heteronuclear NMR spectroscopy revealed a disordered structure in R2 where the second HTH helix was predicted [Jamin et al. (1993) Eur. J. Biochem., 216, 147-154]. We propose that the disordered region folds into a 'recognition' helix and generates a full HTH-related motif upon binding to DNA. This would move Cys43 into the hydrophobic core of R2. We observed that Cys43 was accessible to N-ethylmaleimide alkylation in the free protein, but inaccessible in the DNA complex. Mutant proteins with charged (C43D) or polar (C43S) side chains in position 43 bound DNA with reduced affinity, while hydrophobic replacements (C43A, C43V and C43I) gave unaltered or improved DNA-binding. Specific DNA-binding enhanced protease resistance dramatically. Fluorescence emission spectra and quenching experiments supported a DNA-induced conformational change. Moreover, reversible oxidation of Cys43 had an effect similar to the inactivating C43D mutation. The highly oxidizable Cys43 could function as a molecular sensor for a redox regulatory mechanism turning specific DNA-binding on or off by controlling the DNA-induced conformational change in R2. PMID- 8223473 TI - Mutations of the intronic IgH enhancer and its flanking sequences differentially affect accessibility of the JH locus. AB - To investigate the role of intronic immunoglobulin heavy chain (IgH) enhancer (E mu) in generating accessibility of the JH locus for VDJ recombination, we generated ES cells in which E mu or its flanking sequences were mutated by replacement with or insertion of an expressed neor gene. Heterozygous mutant ES cells were used to generate chimeric mice from which pre-B cell lines were derived by transformation of bone marrow cells with Abelson murine leukemia virus (A-MuLV). Comparison of the rearrangement status of the normal and mutated alleles in individual pre-B cell lines allowed us to assay for cis-acting effects of the mutations. Replacement of a 700 bp region immediately downstream from the core E mu [which includes part of the 3' matrix associated region (MAR) and the I mu exon] had no obvious effect on rearrangement of the targeted allele, indicating that insertion of a transcribed neor gene into the JH-C mu intron does not affect JH accessibility. In contrast, replacement of an overlapping 1 kb DNA fragment that contains the E mu resulted in a dramatic cis-acting inhibition of rearrangement, demethylation and germline transcription of the associated JH locus. Surprisingly, insertion of the neor gene into the 5' MAR sequence approximately 100 bp upstream of the core E mu also dramatically decreased recombination of the linked JH locus; but, in many lines, did not prevent demethylation of this locus. We conclude that integrity of the E mu and upstream flanking sequences is required for efficient rearrangement of the JH locus and that demethylation of this locus, per se, does not necessarily make it a good substrate for VDJ recombination. PMID- 8223474 TI - Mutations in yeast HAP2/HAP3 define a hybrid CCAAT box binding domain. AB - We describe a detailed genetic analysis of the DNA-binding regions in the HAP2/HAP3 CCAAT-binding heteromeric complex. The DNA-binding domain of HAP2 is shown to be a 21 residue region containing three critical histidines and three critical arginines. Mutation of an arginine at position 199 to leucine alters the DNA-binding specificity of the complex to favor CCAAC over CCAAT. Residues in HAP3 that are critical for DNA-binding comprise a short, seven amino acid region. Three different mutations in the HAP2 DNA-binding domain are suppressed by a mutation in the HAP3 DNA-binding domain. This HAP3 mutation also suppresses mutations in a different region of HAP2 which promotes subunit assembly of the complex. These findings suggest that short regions of HAP2 and HAP3 comprise a hybrid DNA-binding domain and that this domain can help hold the two subunits together in the CCAAT-binding complex. PMID- 8223475 TI - Transcription enhancer factor-1 (TEF-1) DNA binding sites can specifically enhance gene expression at the beginning of mouse development. AB - In an effort to identify transcriptional elements that are recognized at different stages of early mouse development, polyomavirus (PyV) enhancer mutations were selected for their ability to support PyV transcription and replication in various mouse undifferentiated embryonal carcinoma (EC) and embryonic stem (ES) cell lines. Several of these enhancer mutations were then isolated, sequenced and tested for their ability to stimulate the PyV early gene promoter in plasmid DNA that was either transfected into EC, ES and fibroblast cell lines, or injected into the nuclei of mouse 1-cell and 2-cell embryos. EC, ES and fibroblast cell lines showed clear preferences for different enhancer configurations, and cleavage-stage embryos (2- to 8-cell stage) strongly preferred the same enhancer configuration favored by ES cells. This 'embryo responsive' (ER) enhancer configuration was characterized by a tandem duplication of the region containing a single point mutation that created a DNA binding site for Transcription Enhancer Factor-1 (TEF-1). ER enhancers stimulated the PyV promoter up to 350-fold in embryos, and were up to 74-fold more active than the wild-type PyV enhancer. Most of the activity from PyER enhancers could be duplicated in 2-cell embryos by synthesizing only the tandemly repeated sequence. Comparison of these synthetic enhancers with ER enhancers confirmed that TEF-1 DNA binding sites were highly preferred in ES cells and cleavage-stage embryos, and suggested that ER enhancer activity resulted primarily from cooperative interaction between either two closely spaced TEF-1 DNA binding sites or two TEF 1 DNA binding sites separated by a third, as yet unidentified, transcription factor binding site. These results provide a prototype of a mammalian embryo responsive enhancer, and suggest that TEF-1 plays an important role in activation of gene expression at the beginning of mammalian development. PMID- 8223476 TI - LEF-1 contains an activation domain that stimulates transcription only in a specific context of factor-binding sites. AB - Lymphoid enhancer factor 1 (LEF-1) is a member of the high mobility group (HMG) family of proteins and participates in the regulation of the T cell receptor (TCR) alpha enhancer. We have previously shown that DNA binding by the HMG domain of LEF-1 induces a sharp bend in the DNA helix. Together with the dependence of LEF-1 on other factor-binding sites to regulate gene expression, DNA bending induced by the HMG domain suggested an 'architectural' role for LEF-1. In this study, we performed experiments to distinguish between a model in which the HMG domain is the only functional determinant of LEF-1 and a model in which additional domains of LEF-1 are involved in the regulation of gene expression. First, we show that the HMG domain alone is not sufficient to stimulate TCR alpha enhancer function. Second, we replaced the HMG domain of LEF-1 with the DNA binding domain of the bacterial repressor LexA, which binds a specific nucleotide sequence without inducing a sharp bend in the DNA helix. The chimeric LEF-LexA protein increased the activity of a TCR alpha enhancer in which the LEF-1-binding site had been replaced with a LexA recognition sequence. Transcriptional stimulation by LEF-LexA, however, was less efficient than that observed with endogenous LEF-1. The LEF-LexA-mediated activation of gene expression was dependent upon an amino-terminal region of LEF-1 and a specific context of factor binding sites in the TCR alpha enhancer. Neither multimerized LexA-binding sites, nor TCR alpha enhancers with altered spatial arrangements of factor-binding sites, were functional for regulation by LEF-LexA. Together, these data suggest that an aminoterminal region in LEF-1 contributes to the context-dependent regulation of the TCR alpha enhancer by LEF-1, presumably by interacting with other enhancer-bound proteins. PMID- 8223477 TI - Synthesis of reinitiated transcripts by mammalian RNA polymerase II is controlled by elongation factor SII. AB - Previous studies have revealed that the in vitro synthesis of reinitiated transcripts by RNA polymerase II requires an additional activity, designated reinitiation transcription factor (RTF), which is distinct from all of the general class II initiation factors. While further characterizing this activity, it was found that RTF displays properties indistinguishable from those of the RNA polymerase II elongation factor SII. In addition, Western blot analysis using SII specific antibodies revealed that human SII is a major component in purified RTF preparations. The functional equivalence of the two proteins was established using recombinant SII, which proved fully capable of substituting for RTF in the reinitiation assay. In these reconstituted reactions, transcription complexes resulting from reinitiation events required SII to proceed through a 400 bp G free cassette, while complexes resulting from the first round of initiations were SII-independent. Reinitiations can take place in the absence of SII; however, addition of the elongation factor is essential for full extension of the reinitiated transcripts. These results suggest that events taking place at the promoter (e.g. first-round initiations versus reinitiations) can create marked differences in the properties of RNA polymerase II elongation complexes. PMID- 8223479 TI - In vitro genetic analysis of the structural features of the pre-tRNA required for determination of the 3' splice site in the intron excision reaction. AB - During processing of intron-containing pre-tRNAs, the Xenopus laevis splicing endonuclease binds the precursor and cleaves it at both the 5' and 3' splice sites. In vitro selection was used to determine structural features characteristic of precursor tRNA molecules that are active in this reaction. We performed two types of selection, one for molecules that are not cut, the other for molecules that are cut at only one site. The results shed light on various aspects of the intron excision reaction, including the importance of the three dimensional structure of the mature domain for recognition and binding of the enzyme, the active role played by the single-stranded region of the intron, and the importance of the cardinal positions which, although not necessarily occupied by the same base in all precursors, nevertheless play a fundamental role in the splicing reaction. A precursor can be cut at the 3' site if a base in the single stranded loop of the intron is allowed to pair (A-I pair) with the base of the 5' exon situated at the position immediately following the anticodon stem [first cardinal position (CP1)]. The nature of the bases involved in the A-I pair is important, as is the position of the base in the single-stranded loop of the intron. We discuss the role of the cardinal positions in the reaction. PMID- 8223478 TI - In vivo control of NF-kappa B activation by I kappa B alpha. AB - The transcription factor NF-kappa B is stored in the cytoplasm in complexes with the inhibitor protein I kappa B alpha. It has been shown in vitro that dissociation of I kappa B alpha from these complexes results in active NF-kappa B. In this report we show that lipopolysaccharide (LPS)-induced activation of B or pre-B cells results in loss of I kappa B alpha from NF-kappa B complexes in vivo. Many liberated NF-kappa B dimers reached the nucleus, where increased c rel, p65 and p50 were detected by immunoblotting and by DNA binding assays. Some liberated dimers were retained in the cytoplasm, however, through binding to newly synthesized I kappa B alpha, a finding which strongly suggests (i) that the LPS-induced signal causes dissociation of complexes rather than preventing their association and (ii) that dissociation results from modification of I kappa B alpha and not of c-rel or p65. No effect of LPS treatment was detected on p105 or p100, which also retain rel family members in the cytoplasm. Quite unexpectedly, we also found that in unstimulated cells there is a constant ongoing process of degradation and replacement of complexed I kappa B alpha. We propose that this turnover results in the low level of active NF-kappa B presumably necessary even in the unstimulated cell, and that the high rate of synthesis of I kappa B alpha provides the ability to turn off NF-kappa B activity rapidly as soon as the activating signal ceases. PMID- 8223480 TI - Functional analysis of pre-mRNA splicing factor SF2/ASF structural domains. AB - Human pre-mRNA splicing factor SF2/ASF has an activity required for general splicing in vitro and promotes utilization of proximal alternative 5' splice sites in a concentration-dependent manner by opposing hnRNP A1. We introduced selected mutations in the N-terminal RNA recognition motif (RRM) and the C terminal Arg/Ser (RS) domain of SF2/ASF, and assayed the resulting recombinant proteins for constitutive and alternative splicing in vitro and for binding to pre-mRNA and mRNA. Mutants inactive in constitutive splicing can affect alternative splice site selection, demonstrating that these activities involve distinct molecular interactions. Specific protein-RNA contact mediated by Phe56 and Phe58 in the RNP-1 submotif of the SF2/ASF RRM are essential for constitutive splicing, although they are not required for RRM-mediated binding to pre-mRNA. The RS domain is also required for constitutive splicing activity and both Arg and Ser residues are important. Analysis of domain deletion mutants demonstrated strong synergy between the RRM and a central degenerate RRM repeat in binding to RNA. These two domains are sufficient for alternative splicing activity in the absence of an RS domain. PMID- 8223481 TI - Functional domains of the human splicing factor ASF/SF2. AB - The human splicing factor ASF/SF2 displays two predominant activities in in vitro splicing assays: (i) it is an essential factor apparently required for all splices and (ii) it is able to switch utilization of alternative 5' splice sites in a concentration-dependent manner. ASF/SF2 is the prototype of a family of proteins typified by the presence of one or two RNP-type RNA binding domains (RBDs) and a region highly enriched in repeating arginine-serine dipeptides (RS regions). Here we describe a functional analysis of ASF/SF2, which defines several regions essential for one, or both, of its two principal activities, and provides insights into how this type of protein functions in splicing. Two isoforms of the protein, which arise from alternative splicing, are by themselves inactive, but each can block the activity of ASF/SF2, thereby functioning as splicing repressors. Some, but not all, mutations in the RS region prevent ASF/SF2 from functioning as an essential splicing factor. However, the entire RS region can be deleted without reducing splice site switching activity, indicating that it is not absolutely required for interaction with other splicing factors. Experiments with deletion and substitution mutants reveal that the protein contains two related, but highly diverged, RBDs, and that both are essential for activity. Each RBD by itself retains the ability to bind RNA, although optimal binding requires both domains. PMID- 8223482 TI - Independent expression of the transforming amino-terminal domain of SV40 large I antigen from an alternatively spliced third SV40 early mRNA. AB - We found that simian virus 40 (SV40), in addition to the SV40 early proteins large T antigen (large T) and small antigen (small t), codes for a third early protein with a molecular weight of 17 kDa. This protein (17kT) is expressed from an alternatively spliced third SV40 early mRNA, using a splice donor site at position 4425 and a splice acceptor site at position 3679 of the SV40 genome. The 17kT protein consists of 135 amino acids. Of these, 131 correspond to the amino terminus of large T, while the four carboxy-terminal amino acids are unique and encoded by a different reading frame. 17kT mRNA, and the corresponding protein, were found in all SV40 transformed cells analyzed, as well as in SV40 infected cells. Transfection of a cDNA expression vector encoding the 17kT protein into rat F111 fibroblasts induced phenotypic transformation of these cells. The expression of the transforming amino-terminal domain of large T as an independent 17kT protein might provide a means for individually regulating the various functions associated with this domain. PMID- 8223483 TI - Methylation interference experiments identify bases that are essential for distinct catalytic functions of a group I ribozyme. AB - Methylation interference experiments reveal bases involved in three different catalytic functions of the T4-phage derived sunY self-splicing intron. RNA molecules methylated at the N-7 position of the guanine at the cofactor binding site are inactive in cofactor-dependent splicing and 3' splice-site hydrolysis. In contrast, 5' splice-site hydrolysis occurs despite methylation at this position. Specific adenines that have been implicated in docking of the P1 stem to the catalytic core are shown to be important for cofactor-dependent splicing and essential for 5' splice-site hydrolysis. Similarly, methylation of bases in the P9.0 stem, as well as C56 in J5/4, interferes with 3' splice-site hydrolysis and with the splicing reaction. All of the bases identified as important for the overall splicing reaction are also identified as essential for either the 5' or 3' splice-site hydrolysis reactions, and vice versa. It is inferred that the bases implicated in 5' and 3' splice-site hydrolysis are involved in specific interactions of the 5' and 3' splice site, respectively, with the catalytic core. PMID- 8223484 TI - Quality control of ER synthesized proteins: an exposed thiol group as a three-way switch mediating assembly, retention and degradation. AB - Plasma cells secrete IgM only in the polymeric form: the C-terminal cysteine of the mu heavy chain (Cys575) is responsible for both intracellular retention and assembly of IgM subunits. Polymerization is not quantitative, and part of IgM is degraded intracellularly. Neither chloroquine nor brefeldin A (BFA) inhibits degradation, suggesting that this process occurs in a pre-Golgi compartment. Degradation of IgM assembly intermediates requires Cys575: the monomeric IgMala575 mutant is stable also when endoplasmic reticulum (ER) to Golgi transport is blocked by BFA. Addition of the 20 C-terminal residues of mu to the lysosomal protease cathepsin D is sufficient to induce pre-Golgi retention and degradation of the chimeric protein: the small amounts of molecules which exit from the ER are mostly covalent dimers. By contrast, when retained by the KDEL sequence, cathepsin D is stable in the ER, indicating that retention is not sufficient to cause degradation. Replacing the C-terminal cysteine with serine restores transport through the Golgi. As all chimeric cathepsin D constructs display comparable protease activity in vitro, their different fates are not determined by gross alterations in folding. Thus, also out of its normal context, the mu chain Cys575 plays a crucial role in quality control, mediating assembly, retention and degradation. PMID- 8223485 TI - Identification and localization of ERD2 in the malaria parasite Plasmodium falciparum: separation from sites of sphingomyelin synthesis and implications for organization of the Golgi. AB - The ERD2 gene product in mammalian cells and yeast is a receptor required for protein retention in the endoplasmic reticulum (ER); immunolocalization studies indicate that the protein is concentrated in the cis Golgi. We have identified a homologue of ERD2 in the malaria parasite, Plasmodium falciparum (PfERD2). The deduced protein sequence is 42% identical to mammalian and yeast homologues and bears striking homology in its proposed tertiary structure. PfERD2 is tightly confined to a single focus of staining in the perinuclear region as seen by indirect immunofluorescence. This is redistributed by brefeldin A (BFA) to a diffuse pattern similar to that of parasite BiP, a marker for the ER; removal of the drug results in recovery of the single focus, consistent with the localization of PfERD2 to the parasite Golgi and its participation in a retrograde transport pathway to the ER. Sphingomyelin synthesis is a second resident activity of the cis Golgi whose organization is sensitive to BFA in mammalian cells. Within the parasite it again localizes to a perinuclear region but does not reorganize upon BFA treatment. The results strongly suggest that these two activities are in distinct compartments of the Golgi in the malaria parasite. PMID- 8223486 TI - Phospholipid transfer activity is relevant to but not sufficient for the essential function of the yeast SEC14 gene product. AB - To investigate several key aspects of phosphatidylinositol transfer protein (PI TP) function in eukaryotic cells, rat PI-TP was expressed in yeast strains carrying lesions in SEC14, the structural gene for yeast PI-TP (SEC14p), whose activity is essential for Golgi secretory function in vivo. Rat PI-TP expression effected a specific complementation of sec14ts growth and secretory defects. Complementation of sec14 mutations was not absolute as rat PI-TP expression failed to rescue sec14 null mutations. This partial complementation of sec14 lesions by rat PI-TP correlated with inability of the mammalian protein to stably associate with yeast Golgi membranes and was not a result of rat PI-TP stabilizing the endogenous sec14ts gene product. These collective data demonstrate that while the in vitro PI-TP activity of SEC14p clearly reflects some functional in vivo property of SEC14p, the PI-TP activity is not the sole essential activity of SEC14p. Those data further identify an efficient Golgi targeting capability as a likely essential feature of SEC14p function in vivo. Finally, the data suggest that stable association of SEC14p with yeast Golgi membranes is not a simple function of its lipid-binding properties, indicate that the amino-terminal 129 SEC14p residues are sufficient to direct a catalytically inactive form of rat PI-TP to the Golgi and provide the first evidence to indicate that a mammalian PI-TP can stimulate Golgi secretory function in vivo. PMID- 8223487 TI - Peroxisomes in the methylotrophic yeast Hansenula polymorpha do not necessarily derive from pre-existing organelles. AB - We have identified two temperature-sensitive peroxisome-deficient mutants of Hansenula polymorpha (ts6 and ts44) within a collection of ts mutants which are impaired for growth on methanol at 43 degrees C but grow well at 35 degrees C. In both strains peroxisomes were completely absent in cells grown at 43 degrees C; the major peroxisomal matrix enzymes alcohol oxidase, dihydroxyacetone synthase and catalase were synthesized normally but assembled into the active enzyme protein in the cytosol. As in wild-type cells, these enzymes were present in peroxisomes under permissive growth conditions (< or = 37 degrees C). However, at intermediate temperatures (38-42 degrees C) they were partly peroxisome-bound and partly resided in the cytosol. Genetic analysis revealed that both mutant phenotypes were due to monogenic recessive mutations mapped in the same gene, designated PER13. After a shift of per13-6ts cells from restrictive to permissive temperature, new peroxisomes were formed within 1 h. Initially one--or infrequently a few--small organelles developed which subsequently increased in size and multiplied by fission during prolonged permissive growth. Neither mature peroxisomal matrix nor membrane proteins, which were present in the cytosol prior to the temperature shift, were incorporated into the newly formed organelles. Instead, these proteins remained unaffected (and active) in the cytosol concomitant with further peroxisome development. Thus in H.polymorpha alternative mechanisms of peroxisome biogenesis may be possible in addition to multiplication by fission upon induction of the organelles by certain growth substrates. PMID- 8223488 TI - The alpha 1 beta 1 integrin recognition site of the basement membrane collagen molecule [alpha 1(IV)]2 alpha 2(IV). AB - Cells interact with type IV collagen mainly via the integrins alpha 1 beta 1 and alpha 2 beta 1. A triple helical CNBr derived fragment CB3[IV], which contains the recognition sites for both integrins, was isolated from type IV collagen. Trypsin treatment of CB3[IV] gave rise to four smaller fragments, F1-F4, of which the smallest one, F4, contained the recognition site for alpha 1 beta 1. Further fragmentation of F4 by thermolysin treatment at 50 degrees C led to fragment TL1, which represents the C-terminal half of F4, and which was no longer able to interact with alpha 1 beta 1. Therefore the recognition site of alpha 1 beta 1 had to be located within the N-terminal half of F4, a position which was verified by electron micrographs of a crosslinked F2-alpha 1 beta 1 complex. Modification of the Arg and Asp residues, which abolished the binding activity of F4, led to the identification of Arg (461) within the alpha 2(IV) and Asp (461) within the alpha 1 (IV) chain as essential residues for the alpha 1 beta 1. The array of these two residues on the surface of the triple helix is discussed. PMID- 8223489 TI - Cell wall sorting signals in surface proteins of gram-positive bacteria. AB - Staphylococcal protein A is anchored to the cell wall, a unique cellular compartment of Gram-positive bacteria. The sorting signal sufficient for cell wall anchoring consists of an LPXTG motif, a C-terminal hydrophobic domain and a charged tail. Homologous sequences are found in many surface proteins of Gram positive bacteria and we explored the universality of these sequences to serve as cell wall sorting signals. We show that several signals are able to anchor fusion proteins to the staphylococcal cell wall. Some signals do not sort effectively, but acquire sorting activity once the spacing between the LPXTG motif and the charged tail has been increased to span the same length as in protein A. Thus, signals for cell wall anchoring in Gram-positive bacteria are as universal as signal (leader) sequences. PMID- 8223490 TI - Nuclear Ca2+ concentration measured with specifically targeted recombinant aequorin. AB - Activation of nuclear transcription factors, breakdown of nuclear envelope and apoptosis represent a group of nuclear events thought to be modulated by changes in nucleoplasmic Ca2+ concentration, [Ca2+]n. Direct evidence for, or against, this possibility has been, however, difficult to obtain because measurements of [Ca2+]n are hampered by major technical problems. Here we describe a new approach for selectively monitoring Ca2+ concentrations inside the nucleus of living cells, which is based on the construction of a chimeric cDNA encoding a fusion protein composed of the photoprotein aequorin and a nuclear translocation signal derived from the rat glucocorticoid receptor. This modified aequorin (nuAEQ), stably expressed in HeLa cells, was largely confined to the nucleoplasm and thus utilized for monitoring [Ca2+]n in intact cells. No significant differences were observed between [Ca2+]n and cytosolic Ca2+ concentration ([Ca2+]i) under resting conditions. Upon stimulation of surface receptors linked to inositol-1,4,5 trisphosphate (InsP3) generation, and thus to intracellular Ca2+ signalling, the kinetics of [Ca2+]i and [Ca2+]n increases were indistinguishable. However, for the same rise in [Ca2+]i, the amplitude of [Ca2+]n increase was larger when evoked by Ca2+ mobilization from internal stores than when induced by Ca2+ influx across the plasma membrane. The functional significance of these transient nucleus-cytosol Ca2+ gradients is discussed. PMID- 8223491 TI - Antibacterial and haemolytic peptides containing D-alloisoleucine from the skin of Bombina variegata. AB - A family of bombinin-related peptides is present in the skin of Bombina variegata. These peptides contain 27 residues with Gly as N-terminus and display antimicrobial activity. From sequence analysis of the cDNAs encoding for the corresponding peptide precursors, the presence of a novel 20-residue peptide with Ile as N-terminus was predicted. We have now purified a family of hydrophobic peptides named H1-H5, whose sequences correspond to the predicted peptide with some variability in positions 1, 2 and 8. In particular, H3-H5 contain a D alloisoleucine residue in the second position. All these peptides display antibacterial and haemolytic activity. PMID- 8223492 TI - Drosophila protein phosphatase V functionally complements a SIT4 mutant in Saccharomyces cerevisiae and its amino-terminal region can confer this complementation to a heterologous phosphatase catalytic domain. AB - The sequence of a Drosophila melanogaster cDNA encoding a novel 35 kDa protein serine/threonine phosphatase, termed PPV, is presented. PPV is 40-41% identical to Drosophila PP1, 53% identical to Drosophila PP2A and 63% identical to Saccharomyces cerevisiae SIT4. Complementation studies demonstrated that PPV can functionally rescue a temperature sensitive mutant of SIT4, a protein phosphatase required for the G1 to S transition of the cell cycle. When placed under the SIT4 promoter, PPV cDNA is able to replace the SIT4 gene in S. cerevisiae. The amino terminal domain of PPV fused to another phosphatase catalytic region (PP1) also rescues the temperature sensitive SIT4 mutant and the SIT4 deletion mutant, implicating this region in binding to regulatory subunits and/or altering specificity. In Drosophila, a substantial transient increase in both PPV mRNA and protein occurs in late syncytial and early cellular blastoderm embryos. At the latter stage PPV is localized to the cytoplasm of cells at the cortex. This increase in PPV correlates with introduction of the G2 phase of the cell cycle, elevated zygotic transcription and cellularization, indicating that PPV may play a role in one or more of these processes. PMID- 8223493 TI - Calpain-catalyzed cleavage and subcellular relocation of protein phosphotyrosine phosphatase 1B (PTP-1B) in human platelets. AB - The non-transmembrane phosphotyrosine phosphatase 1B (PTP-1B) is an abundant enzyme, normally localized to the cytosolic face of the endoplasmic reticulum via a C-terminal targeting sequence. We have found that agonist-induced platelet activation results in proteolytic cleavage of PTP-1B at a site upstream from this targeting sequence, causing subcellular relocation of its catalytic domain from membranes to the cytosol. PTP-1B cleavage is catalyzed by the calcium-dependent neutral protease calpain and is a general feature of platelet agonist-induced aggregation. Moreover, PTP-1B cleavage correlates with the transition from reversible to irreversible platelet aggregation in platelet-rich plasma. Engagement of gpIIb-IIIa is necessary for inducing PTP-1B cleavage, suggesting that integrins regulate tyrosine phosphatases as well as tyrosine kinases. PTP-1B cleavage is accompanied by a 2-fold stimulation of its enzymatic activity, as measured by immune complex phosphatase assay, and correlates with discrete changes in the pattern of tyrosyl phosphorylation. Cleavage and subcellular relocation of PTP-1B represents a novel mechanism for altering tyrosyl phosphorylation that may have important physiological implications in cell types other than platelets. PMID- 8223494 TI - Dephosphorylation of photosystem II core proteins is light-regulated in vivo. AB - A number of photosystem II (PSII)-associated proteins, including D1, D2, CP43 and LHCII, are phosphorylated post-translationally by a membrane-bound, redox regulated kinase activity. In vitro studies have demonstrated that these proteins can be dephosphorylated by membrane-bound phosphatase activity, reportedly insensitive to light or redox control. We demonstrate here that the PSII core proteins, D1, D2 and CP43, undergo light-stimulated, linear electron-transport independent dephosphorylation in vivo. The in vivo dephosphorylation of D1 was characterized further and shown to depend upon light intensity, and to occur throughout the visible light spectrum with characteristics most consistent with light absorption by chlorophyll. PSII core protein dephosphorylation in vivo was stimulated by photosystem I (PSI)-specific far-red light, and inhibited by 2,5 dibromo-3-methyl-6-isopropyl-p-benzoquinone, an inhibitor of plastoquinol oxidation by the cytochrome b6f complex. Based on these findings, we propose that PSI excitation is involved in regulating dephosphorylation of PSII core proteins in vivo. PMID- 8223495 TI - Abnormal T cell development in CD3-zeta-/- mutant mice and identification of a novel T cell population in the intestine. AB - The T cell antigen receptor (TCR)-associated invariable membrane proteins (CD3 gamma, -delta, -epsilon and -zeta) are critical to the assembly and cell surface expression of the TCR/CD3 complex and to signal transduction upon engagement of TCR with antigen. Disruption of the CD3-zeta gene by homologous recombination resulted in a structurally abnormal thymus which primarily contained CD4- CD8- and TCR/CD3very lowCD4+CD8+ cells. Spleen and lymph nodes of CD3-zeta-/- mutant mice contained a normal number and ratio of CD4+ and CD8+ single positive cells that were TCR/CD3very low. These splenocytes did not respond to antibody cross linking or mitogenic triggering. The V beta genes of CD4-CD8- and CD4+CD8+ thymocytes and splenic T cells were productively rearranged. These data demonstrated that (i) in the absence of the CD3-zeta chain, the CD4- CD8- thymocytes could differentiate to CD4+CD8+ TCR/CD3very low thymocytes, (ii) that thymic selection might have occurred, (iii) but that the transition to CD4+CD8- and CD4-CD8+ cells took place at a very low rate. Most strikingly, intraepithelial lymphocytes (IELs) isolated from the small intestine or the colon expressed normal levels of TCR/CD3 complexes on their surface which contained Fc epsilon RI gamma homodimers. In contrast to CD3-zeta containing IELs, these cells failed to proliferate after triggering with antibody cross-linking or mitogen. In comparison to thymus-derived peripheral T cells in the spleen and lymph nodes, the preferential expression of normal levels of TCR/CD3 in intestinal IELs suggested they mature via an independent extrathymic pathway. PMID- 8223496 TI - The C-terminal helix in subdomain 4 of the regulatory light chain is essential for myosin regulation. AB - In vertebrate smooth/non-muscle myosins, phosphorylation of the regulatory light chains by a specific calmodulin-activated kinase controls both myosin head interaction with actin and assembly of the myosin into filaments. Previous studies have shown that the C-terminal domain of the regulatory light chain is crucial for the regulation of these myosin functions. To further dissect the role of this region of the light chain in myosin regulation, a series of chicken smooth muscle myosin regulatory light chain mutants has been constructed with successive C-terminal deletions. These mutants were synthesized in Escherichia coli and analysed by their ability to restore Ca2+ regulation to scallop myosin that had been stripped of its native regulatory light chains ('desensitized'). The results show that regulatory light chain mutants with deletions in the C terminal helix in subdomain 4 were able to reform the regulatory Ca2+ binding site on the scallop myosin head, but had lost the ability to suppress scallop myosin filament assembly and interaction with actin in the absence of Ca2+. Further deletions in the C-terminal domain led to a gradual loss of ability to restore the regulatory Ca2+ binding site. Thus, the regions in the C-terminal half of the regulatory light chain responsible for myosin regulation can be identified. PMID- 8223498 TI - The Brachyury gene encodes a novel DNA binding protein. PMID- 8223497 TI - Novel gene expression mechanism in a fission yeast retroelement: Tf1 proteins are derived from a single primary translation product. AB - In sharp contrast to the single ORF of the Schizosaccharomyces pombe retrotransposon Tf1, retroviruses and most retrotransposons employ two different ORFs to separately encode the Gag and Pol proteins. The different ORFs are thought to allow for overexpression of the Gag protein relative to Pol protein presumed necessary for the assembly of functional retrovirus particles and virus like particles (VLPs). The results of in vivo experiments designed to detect the transposition of Tf1 show that Tf1 is indeed active and can insert itself into the host genome via a true retrotransposition process. Thus, a paradox emerged between the lack of any obvious means of overexpressing Tf1 Gag protein and the demonstrated functionality of the element. Epitope tagging experiments described here confirm that the Tf1 large ORF is intact and that there is no translational or transcriptional mechanism used to overexpress the Tf1 Gag protein. In addition, we used sucrose gradients and antisera specific for Tf1 capsid (CA) and integrase (IN) to show that the Tf1 proteins do assemble into uniform populations of macromolecular particles that also cosediment with Tf1 reverse transcription products. This evidence suggests that Tf1 proteins form VLPs without using the previously described mechanisms that retroviruses and retrotransposons require to overexpress Gag proteins. PMID- 8223499 TI - Update on target theory as applied to chromosomal aberrations. AB - The early radiobiologists, who developed target theory to explain their results, considered the chromosome "target" as a visible thread that could be physically broken by ionizing radiation. Most of the broken ends restituted, but those that did not were free to wander about and, within limits, could rejoin with any other broken end they happened to contact to form structural aberrations. Failing this, they could remain to be seen as "open" breaks at the subsequent metaphase. These ideas, and their inevitable consequences, still form the basis for much modern thinking, even though we now known that the structure of the chromosome, and of the interphase nucleus, are very much more complicated than the originators of the theory envisaged. Current understanding of chromosomes at the molecular level and the varied responses a cell can mobilize when damage is introduced, raise again the question, Can we still think in terms of simple targets? Some of the experimental observations and suggestions made since those early days are reviewed, and the application of target theory to the three theories of aberration origins (Classic, Exchange, Recombination) is briefly discussed. PMID- 8223500 TI - Time sequence of events leading to chromosomal aberration formation. AB - Investigations have been carried out which have measured the influence of the repair polymerases on the yield of different types of chromosomal aberrations. The studies were mainly concerned with the effect of inhibiting the polymerases on the yield of aberrations. The polymerases fill in single strand regions, and the fact that their inhibition affects the yield of aberrations suggests that single strand lesions are influential in aberration formation. The results indicate that-- 1. There are two actions of polymerases in clastogenesis. One is in their involvement in a G2 repair system, in which the pair of chromatids is concerned, and which does not yield aberrations unless the inhibition is still operating when the cells enter mitosis. The second also operates in G1 and S, and is such that when repair is inhibited, further damage accrues. 2. The second action is affected by inhibiting polymerase but operates even when the repair enzymes are active. 3. The production of chromosomal exchanges involves a series of reactions, some of which are reversible. 4. The time span over which the reactions occur is much longer than has been envisaged previously (e.g., most of a cell cycle). PMID- 8223501 TI - DNA double-strand breaks in mutagenesis. PMID- 8223502 TI - Mechanism of chromosome exchange formation in human fibroblasts: insights from "chromosome painting". AB - We have used the techniques of premature chromosome condensation (PCC) and fluorescence in situ hybridization (FISH) with a library for human chromosome 4 to analyze the rate of rejoining of chromosome breaks and development of exchange aberrations in AG1522 human fibroblasts. AG1522 cells were irradiated in plateau phase with 10 Gy and fused with mitotic HeLa cells either immediately after irradiation or at intervals up to eight days later. The slides were then hybridized with the chromosome 4 library and unrejoined breaks and exchange events (visualized as bicolor chromosomes) scored in these cells. At the earliest time point after irradiation, the number of exchange events in the irradiated cells was low, but increased with kinetics similar to that of the joining of the breaks. Furthermore, when we analyzed those cells which had exchange events for their distribution, almost all of the cells initially contained one exchange event (1 bicolor chromosome). As time progressed, the number of cells containing exchanges with two exchange events per cell increased as the number with one exchange event per cell decreased. Extrapolation of the number of exchange events to zero time (with an estimate of 20 min for the fusion and condensation times) gave a value consistent with zero exchanges at zero time after irradiation. In a separate experiment, we also scored AG 1522 cells at the first metaphase after a dose of 6 Gy and were able to show that as many as 50% of the complete exchanges were non-reciprocal in nature, that is, the two broken ends of a single break in chromosome 4 joined to two different chromosomes. These data support the classical breakage-and-reunion model rather than the Revell Exchange Theory of exchange formation. PMID- 8223503 TI - DNA-nuclear matrix interactions and ionizing radiation sensitivity. PMID- 8223504 TI - Interchange and intra-nuclear architecture. AB - For chromatin lesions to interact to form exchanges of any sort, it is obvious that contact between them must be made. However, the probability of such interaction is conditioned by other factors like time, initial separation, metabolic activity, and, in the case of chemically induced lesions, scheduled DNA synthesis. The irradiated nucleus was, for a long time, regarded as a "bag of broken chromosomes" with the severed ends free to move around and find partners with which to form illegitimate reunions. Many of these would be seen at following metaphase as intra- and interchanges. Evidence is rapidly accumulating which indicates that this picture of the nucleus is false. We know now that chromosomes occupy highly localised domains with limited movement, and that there is no massive intermingling; that much of the chromatin is compacted and splinted with proteins and so precluded from exchange-type contact; that most of the chromatin is looped and "fixed" into an intra-nuclear protein scaffold or skeleton; that some chromatin is spun-out and associated with the nuclear envelope in the vicinity of the pore-complexes. Thus it would appear that movement, in the sense envisaged by early workers, is curtailed, and that only a proportion (probably a small proportion) of the chromatin is actually "at-risk" with respect to interchange formation. Where then does interchange take place? Are the "sites" pre-existent, or can proximity requirements be realised after radiation exposure? In what ways will the intra-nuclear architecture influence exchange? These are some of the questions which are considered in this paper. PMID- 8223505 TI - Telomeres and their possible role in chromosome stabilization. AB - The evidence to date generally supports the hypothesis that telomere capping makes chromosome fragments refractory to subsequent rejoining events, but this control may be somewhat relaxed after chromosome breakage. Cell survival requires that the fragments rejoin before metaphase. Unprotected ends such as those produced by DNA damage are subject to degradation, presumably by endogenous cellular exo- and endonucleases. Telomere repeat sequences may be added to broken chromosome ends to protect the ends from further degradation. That telomeric DNA does not always prevent rejoining raises interesting questions as to what constitutes capping, and how rapidly it occurs after DNA damage in relation to chromosome break rejoining. The prevention of degradation and control of rejoining may be mediated by telomere-specific binding proteins, especially the telomere terminal binding protein [Gualberto et al., 1992; Longtine et al., 1989; Price, 1990; Price and Cech, 1989]. Some of these proteins may be involved in scavenging telomeric DNA when the cell senses that chromosomal breaks have occurred. This mechanism is consistent with the observations of Murnane and Yu [1993], who found that a plasmid with telomere sequences was stably integrated in vivo into a chromosome terminal breakpoint lacking telomere repeats. It is also consistent with the high frequency of interstitial telomere sequences observed in normal cells; a history of DNA damage and repair may be recorded by these sequences (Ijdo et al., 1991]. Although chromosome break rejoining is an efficient process in eukaryotic cells, some breaks are never rejoined and can result in terminal deletions and chromatid and isochromatid deletions at metaphase. It is unclear why these breaks are not rejoined, but it may be due to one or more of the following: 1) chance: broken chromosomes are separated, do not approach sufficiently close to one another, and are consequently physically unable to rejoin; 2) a large number of added telomere repeat sequences indicating to the cell that the chromosome has an authentic telomere; 3) some other DNA modification event that protects DNA ends from degradation, e.g., folding back of DNA ends to form a hairpin, as has been implicated in VDJ recombination [Lieber, 1993]. PMID- 8223506 TI - G2 chromatid aberrations: kinetics and possible mechanisms. AB - Chromatid breaks and exchanges are induced by radiation in G2 mammalian cells. Breaks are at a maximum number at about 30 min after irradiation and decrease apparently exponentially with time between irradiation and sampling. Few breaks are observed immediately following exposure, probably as a result of selection of mitotic cells where chromosomes are condensed and there is consequently a lack of time for expression of damage. The change in frequency of breaks with time, from 30 min after radiation exposure and onwards, can be interpreted in two possible ways: either in terms of a repair process or in terms of a change in radiosensitivity through G2. However, our results with an inhibitor of repair of DNA double-strand breaks (ara A) and with "transient hypothermia" which extends the G2 phase, argue for an interpretation based on rejoining of chromatid breaks, possibly reflecting the repair of a subclass of dsb. Data from experiments with irradiated and restriction endonuclease treated radiosensitive mutant rodent lines indicate that enhanced levels of conversion of dsb into chromosomal aberrations may be largely independent of repair rates of bulk dsb. In CHO cells and in human lymphocytes exchanges initially increase rapidly with time and then remain at a constant frequency, supporting the notion of a uniform chromosomal radiosensitivity throughout most of G2 and providing further evidence that the mechanism for mis-joining broken chromatids (leading to exchanges) is different from that for rejoining of chromatoid breaks. Ratios of breaks to exchanges were found to vary in different cell lines and at different times during treatment with inhibitors or at altered temperatures, possibly (in different cell lines) indicating different levels of enzymes involved in misjoining, but suggesting that the mechanisms of chromosomal rejoining and misjoining are independent, at least to some degree. PMID- 8223507 TI - Molecular mechanisms for the induction of chromosomal aberrations in CHO cells heated in S phase. AB - Chromosomal aberrations are induced by heat only when Chinese hamster ovary cells are heated in S phase of the cell cycle. Studies on the hyperthermic inhibition of cellular DNA replication have indicated that four molecular aspects of DNA replication are affected after heating. New replicon initiation and DNA chain elongation are inhibited; the fork displacement rate is very sensitive to heat inactivation; and finally, there is almost a two-fold increase in single-stranded regions in the replicating DNA after heating. From a comparison of these altered processes between S phase cells and heated G1 cells, which do not die from chromosomal aberrations, our current hypothesis involves 3 steps for the chromosomal aberration induction process. The first critical step is the persistent increase of single-stranded regions in the replicating DNA. Then, we hypothesize that the second step is the creation of transient double strand breaks (DSBs) induced at sites opposite these regions by endogenous endonucleases. Finally, the third step requires that improper repair of these DSBs occurs from either nonrepair or misrepair which then leads to the final chromosomal aberrations seen in the first mitosis after treatment. We believe that this 3 step induction process is common for any cytotoxic agent that induces chromosomal aberrations after DNA replication has been inhibited. PMID- 8223508 TI - Mechanism of hyperoxia-induced chromosomal breakage in Chinese hamster cells. AB - Exposure of cell cultures to hyperoxia, i.e., an atmosphere containing more than 20% O2, results in various genotoxic effects. The most prominent effect of hyperoxia is its clastogenicity. In this paper, earlier published data, obtained from research devoted to the mechanism of hyperoxia-induced clastogenesis, are reviewed. In addition, new data are presented concerning the hyperoxia sensitivity of the DNA-repair deficient Chinese hamster cell lines xrs1, irs1, and EM9. None of these ionizing radiation-sensitive mutants showed hypersensitivity to hyperoxia, as measured by chromosomal aberration induction and loss of clonogenic cell survival. From the normal hyperoxia-sensitivity of xrs1, it may be concluded that DNA double strand breaks, of the type that are induced by ionizing radiation, do not play a role in chromosomal aberration formation by hyperoxia. In addition, since xrs1 is hypersensitive to drugs that inhibit topoisomerase II, it seems rather unlikely that exposure to hyperoxia affects topoisomerase II activity. Based on circumstantial evidence we hypothesize that perturbation of poly(ADP-ribose) metabolism may play a critical role in the mechanism of hyperoxia-induced clastogenesis. PMID- 8223509 TI - Evidence for excision repair-dependent and -independent processes in ara C induced chromosome rearrangements in G1 human lymphocytes. AB - 1-beta-D-arabinofuranosylcytosine (ara C) enhances the formation of chromosome rearrangements such as translocations or dicentric chromosomes in G1 cells containing DNA lesions. The formation of rearrangements is hypothesized to be the result of inhibition of excision repair. Ara C has also been known to lead to the formation of chromosome rearrangements in G1 cells in the absence of induced DNA lesions. It is not known whether a common mechanism is involved in these two processes. In the present study, we used excision repair-deficient XP cells to investigate whether excision repair is involved in the formation of chromosome rearrangements in G1 cells which do not contain induced DNA lesions. G0 Lymphocytes from an XP patient were either treated with 4-nitroquinoline-1-oxide (4NQO) or left untreated. Cells were then cultured in the presence of ara C for about 18 h. Aphidicolin (APC), which induces chromosome rearrangements in cells containing 4NQO-induced DNA lesions, was used for comparison. The resulting frequency of dicentrics and rings (dic & ring) was determined at the first mitoses after culture initiation. In 4NQO-pretreated XP cells, the frequency of dic & ring was not increased by post-treatment with ara C or with APC. This result is thought to reflect the absence of excision repair in XP cells. However, normal induction of dic & ring was observed in XP cells not pretreated with 4NQO but treated with ara C. Thus, there seems to be two different processes involved in the induction of G1 rearrangements: excision repair-dependent and excision repair-independent. UV-endonuclease is not involved in excision repair independent rearrangements. PMID- 8223510 TI - Mechanism of induction of chromosomal aberrations by inhibitors of DNA topoisomerases. AB - Evidence is provided that antitopoisomerase drugs induce chromosomal aberrations via the formation of DNA double strand breaks. Camptothecin, an inhibitor of DNA topoisomerase I, produced only chromatid-type aberrations, and in order to be effective it has to be present during the DNA replication. The fact that it induces chromosomal aberrations also in the G2 phase of the cell cycle suggests the presence of a residual DNA synthesis in this phase. A pretreatment with aphidicolin, an inhibitor of DNA polymerases, reduces the camptothecin induced aberrations in the G2 phase, confirming the need of DNA synthesis for the induction of aberrations by the inhibitor of DNA topoisomerase I. PMID- 8223511 TI - Histone shuttle driven by the automodification cycle of poly(ADP ribose)polymerase. AB - In mammalian cells, the incision step of DNA excision repair triggers a dramatic metabolic response in chromatin. The reaction starts with the binding of a zinc finger protein, i.e. poly(ADP-ribose)polymerase to DNA nicks, activation of four resident catalytic activities leading to poly(ADP-ribose) synthesis, conversion of the polymerase into a protein modified with up to 28 variably sized ADP-ribose polymers, and rapid degradation of polymerase-bound polymers by poly(ADP ribose)glycohydrolase. This automodification cycle catalyzes a transient and reversible dissociation of histones from DNA. Shuttling of histones on the DNA allows selected other proteins, such as DNA helicase A and topoisomerase I, to gain access to DNA. Histone shuttling in vitro mimics nucleosomal unfolding/refolding in vivo that accompanies the postincisional steps of DNA excision repair. Suppression of the automodification cycle in mammalian cells prevents nucleosomal unfolding and nucleotide excision repair. PMID- 8223512 TI - Contribution of O6-alkylguanine and N-alkylpurines to the formation of sister chromatid exchanges, chromosomal aberrations, and gene mutations: new insights gained from studies of genetically engineered mammalian cell lines. AB - O6-methyl- and O6-ethylguanine are the major premutagenic and precarcinogenic lesions induced in DNA by monofunctional alkylating agents, albeit formed in minor amounts. The involvement of these lesions in SCE and aberration formation is less clear. We have analyzed the contribution of O6-alkylguanine to SCE and aberration formation, as well as its toxic and point mutation inducing effect in transgenic Chinese hamster ovary (CHO) cell lines that express variable amounts of human O6-methylguanine-DNA methyltransferase (MGMT). Cells that overexpress MGMT (or the bacterial Ada protein) gained resistance to the formation of alkylation-induced SCEs and aberrations, as compared to MGMT deficient cells. A correlation was apparent between the level of protection for SCEs and cell killing, indicating that both phenomena are interrelated. The protective effects were dependent on the level of MGMT expression, the agent used for alkylation, and cell cycle progression. Our data suggest that at least 2 kinds of lesions are responsible for SCE and aberration formation, namely, O6-alkylguanine and one or various N-alkylation products. The probability that O6-methylguanine is converted into cytogenetic effects has been estimated to be about 1:30 for SCEs, and 1:147,000 and 1:22,000 for chromosomal aberrations in the first and second post treatment mitosis, respectively. The induction of SCEs and likely also of aberrations by O6-methylguanine requires two replication cycles and is supposed to involve the formation of secondary DNA lesions. Increased repair of 3 methyladenine and 7-methylguanine in CHO cells that overexpress the N methylpurine-DNA glycosylase (MPG) after transfection with the human MPG-cDNA did not give rise to protection against methylation-induced SCEs and aberrations, probably because of incomplete excision repair. MPG overexpressing cells reacted even more sensitively to methylating agents, suggesting apurinic sites formed as a result of MPG action to be SCE and aberration-inducing lesions. PMID- 8223513 TI - Mutagenicity of methyl methanesulfonate (MMS) in vivo at the Dlb-1 native locus and a lacI transgene. AB - Methyl methanesulfonate (MMS) is an extraordinarily poor mutagen compared to ethylnitrosourea (ENU) or even X-rays. In lung fibroblasts in vivo, MMS has been shown to induce many micronuclei but few, if any, mutations at the hpt locus. We wondered if the lack of mutations might be due to the lack of division and DNA synthesis in fibroblasts in vivo, which would permit substantial time for differential repair of DNA lesions. This idea was tested in the small intestine, a tissue in which the cells are actively dividing. Two loci were examined: a native locus (Dlb-1) which determines the presence or absence of a lectin binding site on the surface of the epithelial cells, and a lacl transgene which controls beta-galactosidase synthesis. Locl mutations were detected after in vitro packaging of DNA isolated from the intestinal epithelium into lambda phage and expression in suitable bacteria. Although the epithelial cells are proliferating, acute treatments produced no significant increase in mutations at either locus. Subacute treatments produced low but significant increases in mutation frequency at both loci. The results confirm that MMS is a far more potent clastogen than it is a mutagen and should be regarded as a super-clastogen in the same manner as ENU is a super-mutagen. The carcinogenicity of MMS is probably the result of its potent clastogenicity rather than its weak activity as a point mutagen. PMID- 8223514 TI - Muscle coordination and choice-reaction time tests as indicators of occupational muscle load and shoulder-neck complaints. AB - The use was explored of psychomotor tests as indicators of the risk of shoulder neck disorders in workers with low-level static loads on the shoulder muscles. Two groups of workers performing office work and light production work were studied. A muscle coordination test with continuous movement of the arm and hand between three target areas and a psychogenic tension test, posing mental demands and with minimal requirement for body movements, aimed to quantify muscle activity in excess of that needed for biomechanical purposes. The electromyogram (EMG) recording of the active trapezius muscle in the muscle coordination test correlated with the median and static EMG values of the vocational (i.e. during the normal work task) trapezius recording both for the office and production workers, but showed no correlation with shoulder-neck complaints. The EMG responses in the psychogenic tension test and of the passive (contralateral) trapezius in the muscle coordination test correlated best with the parameters showing short, spontaneous pauses in the EMG recording of occupational load. For the office workers, but not for the production workers these parameters also correlated with shoulder-neck complaints and the presence of psychosocial problems. Psychomotor tests may thus be useful as indicators of the risk of shoulder-neck complaints in certain occupations, but further experimentation is needed to validate this conclusion. PMID- 8223515 TI - Comparison of blood pressure response to isokinetic and weight-lifting exercise. AB - Brachial arterial pressure, oesophageal pressure, and knee joint angle were monitored in eight untrained young men as they performed bilateral leg-press actions (simultaneous hip and knee extension and ankle plantarflexion) against resistance. Single maximal leg-press actions on an isokinetic device evoked mean peak arterial (systolic/diastolic) pressures of 35.4/26.2 and 34.0/23.4 kPa at lever arm velocities of 0.262 and 1.31 rad.s-1, respectively. The corresponding oesophageal pressures were 13.2 and 10.4 kPa, respectively. Although the peak force was 30% greater, and duration of the action 3-4 times longer at 0.262 than 1.31 rad.s-1, the arterial and oesophageal pressure responses did not differ. On a weight-lifting machine, a set of repetitions [mean (SEM): 11 (3)] to failure at 80-90% one repetition maximum evoked peak arterial pressures of 45.5/32.8 kPa; the corresponding oesophageal pressure was 15.7 kPa. The peak systolic and diastolic pressures observed during weight-lifting were significantly (P < 0.05) higher than during isokinetic actions at both velocities, whereas oesophageal pressure was more elevated only in relation to isokinetic actions at the higher velocity. These data indicate that resisted leg-press actions cause extreme elevations in arterial blood pressure. The degree of voluntary effort is the major determinant of the blood pressure response, rather than the resistance mode or the type (concentric, eccentric, isometric) of muscle action. Repetitive resistance exercise (e.g. a set of repetitions to failure in weight-lifting) tends to produce greater pressure elevations than isolated, single maximal effort actions. PMID- 8223516 TI - Concentric and eccentric muscle strength before, during and after fatigue in 13 year-old boys. AB - The purpose of this study was to investigate the force-producing characteristics of boys aged 13 years in relation to fatigue of elbow flexor muscles. Maximal voluntary force in elbow flexion was measured before and after a muscle endurance test (MET) by using an isokinetic dynamometer isometrically, concentrically and eccentrically at three velocities, i.e. 0.21, 0.52, and 1.05 rad.s-1. The MET consisted of maximal concentric and eccentric muscle actions performed alternately at 0.52 rad.s-1 for 50 consecutive trials. Muscle cross-sectional area (CSA) of elbow flexor muscles (biceps brachii and brachialis) was measured by a B-mode ultrasound apparatus. Although eccentric force showed significantly higher values than concentric force during MET, there was no significant difference in the rate of decline in force between the two actions. There was no significant difference in the rate of decline in force after MET for each velocity and muscle action. Isometric, concentric and eccentric force before MET was significantly related to muscle CSA whereas, after MET, concentric force significantly correlated with muscle CSA but there was no significant correlation between muscle CSA and isometric or eccentric force. From our study, it is therefore suggested that in development to maturity, isometric, concentric and eccentric force decrease at the same rate with advancing muscle fatigue; however, there might be differences among muscle actions in factors affecting force development. PMID- 8223517 TI - Reliability, reproducibility and validity of the individual anaerobic threshold. AB - The individual anaerobic threshold (IAT) has been defined as the highest metabolic rate at which blood lactate (La) concentrations are maintained at a steady state during prolonged exercise. The validity of this definition, however, has not been substantiated. Eleven men [maximum oxygen uptake (VO2max), mean (SD), 57.8 (6.9) ml.kg-1 x min-1) did two maximal incremental cycle exercise tests (30 W and 4 min per step). Blood was sampled repeatedly during exercise and for 9 min during the subsequent recovery period with light activity. The subjects then exercised at the power output equivalent of IAT for 45 min, until they could no longer continue or until rectal temperature reached 39 degrees C. Subjects performed two additional exercise tests. The intensity of these tests depended upon the LA and acid-base responses during the last 15 min of at least 30 min of exercise at IAT. If a steady state was achieved (La, pH and PCO2 changed by less than 0.5 mmol.l-1, 0.005 pH units and 0.3 kPa, respectively) or decreasing La and increasing pH values were observed, then the second test was performed at IAT +5% VO2max and the third session at either IAT +2.5% or +7.5% VO2max. Conversely if a steady state was not achieved during exercise at the calculated IAT, the intensity of the second test was set at IAT -5% VO2max. Depending on the La and acid-base responses during this test, the final session was performed at either IAT -2.5% or -7.5% VO2max.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223518 TI - Changes in erythrocyte membrane phospholipid composition induced by physical training and physical exercise. AB - The effects were investigated of physical training and exercise on lipids of the erythrocyte membrane of healthy students. Membrane cholesterol and phospholipids were analysed simultaneously by thin-layer chromatography with a flame ionization detector and the fatty acid composition was determined by gas chromatography. Physically trained students had similar physical characteristics to control students but a significantly higher aerobic capacity, estimated as the maximal oxygen uptake and anaerobic threshold. Of the phospholipids examined, only the content of membrane phosphatidylserine was significantly lower in the trained group. Fatty acid analysis showed that the amount of docosahexaenoic acid in membrane phosphatidylserine was lower in the trained group. There was no significant difference between the fatty acid compositions of membrane phosphatidylcholine in the two groups. Maximal exercise decreased membrane phosphatidylserine in the control group but not in the trained group. It also significantly decreased the relative amounts of unsaturated fatty acids in both phosphatidylcholine and phosphatidylserine in the untrained group. Maximal oxygen uptake was negatively correlated with the amount of erythrocyte membrane phosphatidylserine. These results would indicate that both physical training and acute exercise decrease phosphatidylserine and polyunsaturated fatty acids in erythrocyte membranes, possibly due to lipid peroxidation, suggesting limited enhancement of erythrocyte defense mechanisms in adaptation to chronic oxidative stress. PMID- 8223520 TI - Energy cost and efficiency of riding aerodynamic bicycles. AB - Traction resistance (Rt) was determined by towing two cyclists in fully dropped posture on bicycles with an aerodynamic frame with lenticular wheels (AL), an aerodynamic frame with traditional wheels (AT), or a traditional frame with lenticular wheels (TL) in calm air on a flat wooden track at constant speed (8.6 14.6 m.s-1). Under all experimental conditions, Rt increased linearly with the square of air velocity (v2a); r2 equal to greater than 0.89. The constant k = delta Rt/delta v2a was about 15% lower for AL and AT (0.157 and 0.155 N.s2 x m-2) than for TL bicycles (0.184 N.s2 x m-2). These data show firstly, that in terms of mechanical energy savings, the role of lenticular wheels is negligible and, secondly, that for TL bicycles, the value of k was essentially equal to that found by others for bicycles with a traditional frame and traditional wheels (TT). The energy cost of cycling per unit distance (Cc, J.m-1) was also measured for AT and TT bicycles from the ratio of the O2 consumption above resting to speed, in the speed range from 4.7 to 11.1 m.s-1. The Cc also increased linearly with v2a, as described by: Cc = 30.8 + 0.558 v2a and Cc = 29.6 + 0.606 v2a for AT and TT bicycles. Thus from our study it would seem that AT bicycles are only about 5% more economical than TT at 12.5 m.s-1 the economy tending to increase slightly with the speed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223519 TI - Effects of aerobic exercise conditioning at intensities corresponding to lactate threshold in the elderly. AB - In this study we attempted to determine the effects of exercise training at the intensity corresponding to lactate threshold (Thla-) on various health-related variables in sedentary but apparently healthy elderly subjects. Six men and five women volunteers [mean age 68.9 (SD 3.4) years] performed supervised endurance type training on stationary cycle ergometers for 30 min and recreational activities for 30 min, 3 days a week for 12 weeks. Four men and four women served as the control group [68.8 (SD 4.4) years]. As a result of the training programme, statistically significant increases in maximal oxygen consumption (10%), oxygen consumption at Thla- (18%), distance covered in 12-min walk, side step, and leg extensor power were found in the training group, while no changes occurred in the control group. The changes in serum cholesterol and triglyceride concentrations from the pre- to post-training period were statistically significant. High-density lipoprotein cholesterol remained unchanged, and low density lipoprotein cholesterol tended to decrease following the training programme. These data would indicate that exercise training at the intensity corresponding to Thla- may have favourable effects on overall physical fitness and some serum lipid variables in older individuals. PMID- 8223521 TI - Aerobic and anaerobic indices contributing to track endurance cycling performance. AB - A group of 18 male high performance track endurance and sprint cyclists were assessed to provide a descriptive training season specific physiological profile, to examine the relationship between selected physiological and anthropometric variables and cycling performance in a 4000-m individual pursuit (IP4000) and to propose a functional model for predicting success in the IP4000. Anthropometric characteristics, absolute and relative measurements of maximal oxygen uptake (VO2max), blood lactate transition thresholds (Thla- and Th(an),i), VO2 kinetics, cycling economy and maximal accumulated oxygen deficit (MAOD) were assessed, with cyclists also performing a IP4000 under competition conditions. Peak post competition blood lactate concentrations and acid-base values were measured. Although all corresponding indices of Thla- and Th(an),i occurred at significantly different intensities there were high intercorrelations between them (0.51-0.85). There was no significant difference in MAOD when assessed using a 2 or 5 min protocol (61.4 vs 60.2 ml.kg-1, respectively). The highest significant correlations were found among IP4000 and the following: VO2max (ml.kg 2/3.min-1; r = -0.79), power output at lactate threshold (Wthla) (W; r = -0.86), half time of VO2 response whilst cycling at 115% VO2max (s; r = 0.48) and MAOD when assessed using the 5 min protocol (ml.kg-1; r = -0.50). A stepwise multiple regression yielded the following equation, which had an r of 0.86 and a standard error of estimate of 5.7 s: IP4000 (s) = 462.9 - 0.366 x (Wthla) - 0.306 x (MAOD) - 0.438 x (VO2max) where Wthla is in W, MAOD is in ml.kg-1 and VO2max is in ml.kg 1 x min-1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223522 TI - Time of day effects on sympathoadrenal and pressor reactivity to exercise in healthy men. AB - To investigate the influence of time of day on sympathoadrenal and pressor reactivity during exercise, eight trained men [age, mean (SD), 24 (0.5) years; maximal oxygen uptake (VO2max), 4.7 l.min-1] performed bouts of static (ST) and dynamic (DYN) exercise at 0600-0800 hours (AM) and at 1600-1800 hours (PM). The ST protocol utilized a two-leg isometric contraction at 30% maximum voluntary contraction until failure, and was monitored by a strain gauge interfaced from a leg extension apparatus to a computer. Heart rate (fc) and blood pressure (Pa) responses were recorded at rest, after 1 and 2 min of exercise, and at failure. Epinephrine (EPI) and norepinephrine (NE) levels were recorded before exercise, and after 2 min of exercise. The DYN exercise protocol involved stationary cycling for consecutive 6-min periods at 60% and 80% VO2max. fc, Pa, EPI, and NE were recorded before exercise and at each workload. No differences were observed in preexercise or exercise fc under any condition. Preexercise Pa did not differ under any condition. The Pa response to DYN was significantly higher at 80% VO2max during PM only. Pa was significantly higher in ST-PM at 1 min, 2 min, and failure. Elevations in both systolic and diastolic Pa contributed to this difference. Preexercise EPI-ST-AM was significantly elevated vs PM, but no other preexercise data were significantly different. Absolute exercise levels were significantly higher for EPI-ST-PM vs AM only, but the percentage change from baseline was significantly (P < 0.01) higher in ST-PM for EPI (+231% PM vs +32% AM) and NE (+352% PM vs +216% AM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223523 TI - Supine and standing sympathovagal balance in athletes and controls. AB - Differences in autonomic nerve activity between athletes and controls during supine rest and standing were investigated by recording the cardiac rhythm in 18 professional cyclists and 11 controls. We computed four indexes of autonomic control: the standard deviation (SD) of the interbeat intervals, the coefficient of variance (CV) of the interbeat intervals, the percentage of successive intervals differing by more than 50 ms (pNN50), and the fraction low-frequency (0.07-0.14 Hz) spectral power (LF), and we also measured the mean interbeat interval (MI). Significant differences (Student's t-test, P < 0.005) between the athletes and the controls in the supine position were found for pNN50 [mean 52.6 (SEM 2.5) vs 37.1 (SEM 3.4)%], LF [mean 32.2 (SEM 1.6) vs 40.7 (SEM 2.1) normalized units], and MI [mean 1241 (SEM 20) vs 1021 (SEM 25) ms]. A significant difference between the athletes and the controls in the standing position was found for MI [mean 888 (SEM 13) vs 801 (SEM 23) ms]. These results would suggest that there is a parasympathetic predominance in athletes in the supine, but not in the standing position. The finding that pNN50 and LF, but not SD and CV, differed between the athletes and the controls, would seem to demonstrate that the differences in autonomic control between the athletes and the controls are reflected in the quality (balance between slow and fast heart rate fluctuations) rather than in the quantity of heart rate variability. PMID- 8223524 TI - Determination of capillary leakage due to recombinant interleukin-2 by means of noninvasive conductivity measurements. AB - One of the most common side effects of treatment with recombinant interleukin-2 (IL-2) is capillary leakage. Its genesis is not completely understood. The aim of the study was to determine whether capillary leakage can be monitored by means of a noninvasive conductivity technique and to study its starting point. Eight patients with advanced renal cell cancer were studied in a medium care section of the Department of Medical Oncology, University Hospital over 4 days during treatment sessions of continuous, intravenously administered IL-2 (mean dose of 15.6 x 10(6) IU.m-2.day-1). The fluid shift from the intravascular to the extra- and intracellular compartments was monitored by means of noninvasive conductivity measurements. Changes in blood volume were calculated from serial erythrocyte counts. The clinical parameters of capillary leakage (oliguria, positive fluid balance, and gain in mass) were recorded. The mean gain in mass was 9% after 4 days of IL-2 treatment. The extracellular fluid volume increased significantly [46 (SD 23.2)%; P < 0.01], whereas the intracellular fluid volume did not change. The increase in blood volume (BV) amounted to 7% (P < 0.05). The decline in albumin concentration was significantly more than the increase in BV [38 (SD 4.3)%; P < 0.01], indicating capillary albumin leakage. The main changes were observed after the 2nd day of treatment. From this study, it is suggested that conductivity measurements are a suitable method to monitor capillary leakage induced by IL-2, and could be used to detect the exact onset and severity of this leakage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223525 TI - Ratings of perceived exertion and affect in hot and cool environments. AB - The effects of hot and cool environments on perceptual and physiological responses during steady-state exercise were examined in men (n = 14) performing 30 min of constant exercise (cycle ergometry) at a perceived exertion of "somewhat hard". Subjects exercised at the same absolute exercise intensity in hot (40 degrees C), neutral (24 degrees C), and cool (8 degrees C) conditions. Data were collected for differential ratings of perceived exertion (RPE), affect, thermal sensation, mean skin (Tsk) and rectal temperatures (Tre), and cardiac frequency (fc). The subjects completed the hot exposure with an average Tsk of 37.5 degrees C (SEM 0.11), while the neutral and cool conditions produced values of 33.8 (SEM 0.09) and 28.2 degrees C (SEM 0.30), respectively. The Tsk was significantly higher in the hot than the neutral and cool conditions throughout exercise (P < 0.05). The fc was significantly lower in the cool than in the other conditions (P < 0.05), and the subjects completed the hot exposure with a mean fc more than 20 beats.min-1 greater than observed in the other conditions. The subjects felt worse (lower affect) in the heat throughout exercise (P < 0.05). Overall RPE was significantly lower in the cool than in the heat, while chest RPE scores for the cool and hot conditions were displaced vertically by approximately two points. Subjects perceived work to be harder, felt worse, and experienced greater thermal sensation in the hot condition, compared with the neutral and cool conditions. Changes in cutaneous vasomotor tone and heat-induced influences on the chest may have accounted for the RPE changes observed in the heat. PMID- 8223526 TI - Decreased salivary immunoglobulin A secretion rate after intense interval exercise in elite kayakers. AB - Endurance athletes have been shown to suffer a high incidence of upper respiratory tract infection (URTI; e.g. colds, sore throat) during intense training and after competition. Previous studies have shown that concentrations of secretory immunoglobulin A (IgA), the major effector of host defense against micro-organisms causing URTI, decrease after intense endurance exercise. Many athletes perform intense interval exercise as part of their normal training. The purpose of this study was to determine whether salivary IgA concentrations also decrease after intense interval exercise during the normal training regime in elite athletes. Timed saliva samples were obtained from eight elite male kayakers immediately before and after three on-water training sessions during a 3-week period. The concentrations of IgA, IgG and IgM were determined separately by enzyme-linked immunosorbent assay, and secretion rates calculated for each Ig. The IgA secretion rate (micrograms.min-1) decreased 27%-38% after all three training sessions (P = 0.007); the largest decrease (38%) was noted after the most intense session at the end of an especially intense week of training. The IgA concentration relative to total protein (micrograms.mg protein-1) was significantly lower (P < 0.05) on this training day compared with the other 2 days. Concentrations and secretion rates of IgG and IgM did not change after exercise, indicating a specific effect on IgA. These data would suggest that, in elite athletes, IgA concentration and secretion rate are reduced by intense interval exercise, and that exercise-induced changes in IgA output may be one mechanism contributing to URTI in elite athletes. PMID- 8223527 TI - Oxygen deficits incurred during 45, 60, 75 and 90-s maximal cycling on an air braked ergometer. AB - The aims of this study were to determine the most appropriate duration for the measurement of the maximal accumulated O2 deficit (MAOD), which is analogous to the anaerobic capacity, to ascertain the effects of mass, fat free mass (FFM), leg volume (Vleg) and lower body volume (V1b) on anaerobic test performance, to examine the reproducibility for peak power output (Wpeak) or maximal anaerobic power using an air-braked cycle ergometer and to produce approximations for the percentages of aerobic and anaerobic metabolism during exercise of short duration but high intensity. A group of 12 endurance trained cyclists [mean age 25.1 (SD 4.6) years; mean body mass 73.43 (SD 7.12) kg; mean maximal oxygen consumption 5.12 (SD 0.35) l.min-1; mean body fat 12.5 (SD 4.1) %] accordingly performed four counterbalanced treatments of 45, 60, 75 and 90 s of maximal cycling on an air braked ergometer. The mean O2 deficit of 3.52 l for the 45-s treatment was significantly less (P < 0.01) than those for the 60 (3.75 l), 75 (3.80 l) and 90 s (3.75 l) treatments. These data therefore indicate that in predominantly aerobically trained subjects the O2 deficit attains a plateau after 60 s of maximal cycling on an air-braked ergometer. Statistically significant interclass correlation coefficients (P < 0.05) between the anthropometric variables (mass, FFM, Vleg and Vlb) and Wpeak or maximal anaerobic power (0.624-0.748) and MAOD (ml) or anaerobic capacity (0.666-0.772) furthermore would suggest the relevance of taking into account muscle mass during anaerobic tests.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223528 TI - Circadian variation in peripheral blood flow in relation to core temperature at rest. AB - The aim of the present study was to examine the circadian variation in skin blood flow and its relationship to the rhythm of rectal temperature. Eight young men were confined to continuous bed rest for 24 h in a thermoneutral environment. Rectal temperature, oxygen consumption, and peripheral blood flow (laser-Doppler flowmetry and plethysmography) were measured at intervals of 2 h, except at 2400 hours and 0600 hours. According to the cosinor-method, the mean rectal temperature displayed a statistically significant circadian variation with a minimum at 0400 hours. A significant circadian rhythm was also observed in total finger blood flow, finger laser-Doppler flow, and forearm laser-Doppler flow, but not in total forearm blood flow and forehead laser-Doppler flow. The rhythms of rectal temperature and extremity skin blood flow were phased differently, i.e. the decrease in rectal temperature was preceded by an increase in extremity skin blood flow with an average of 4-h time difference. The results would support the idea that circadian rhythm of core temperature is, at least in part, regulated by variation in extremity skin blood flow. PMID- 8223529 TI - Myo-electric signals from two extrinsic hand muscles and force tremor during isometric handgrip. AB - The objective of the present study was to investigate the myo-electric signs of muscle fatigue and the isometric force tremor of two extrinsic hand muscles, extensor digitorum communis and flexor digitorum superficialis, during isometric power grip. In addition, the synergy between flexor and extensor muscles and hand differences in a right-handed population have been studied. During isometric hand dynamometry the myo-electric signal was recorded using surface electrodes and isometric force tremor was recorded using a special load cell. Eight subjects participated in this study and contractions were performed at 20%, 40%, 60% and 80% of maximal voluntary contraction (MVC) with left and right hands. The decrease of mean power frequency (MPF) with duration of contraction was greater in the left extensor as compared to the ipsilateral flexor muscle. No differences in the decrease in MPF with the duration of the contraction were found between the right extensor and flexor muscles. Isometric force tremor root mean square did not change during contractions at a given contraction level. Isometric tremor amplitude increased from 20% to 60% MVC and decreased at higher contraction levels. Tremor amplitude was higher in the left hand at all contraction levels but 60% MVC. These data would suggest differences in fatiguability and muscle fibre composition between the dominant and nondominant hand, which may be due to preferred use. The significance of force tremor for the evaluation of recruitment order and muscle fatigue is discussed. PMID- 8223530 TI - Normalization of electromyogram in the neck-shoulder region. AB - Linear and curvilinear electromyogram (EMG) normalization methods were compared among ten healthy men during a simulated work cycle demanding attention and static holding of the arm ('Solitaire test'). Maximal voluntary contractions (MVC) and gradually increasing contractions up to 70% of MVC were used for normalization in different arm postures. The test contractions studied included inward and outward rotations, abduction, shoulder elevation, and flexion in different arm positions. The shoulder load moment was calculated for the flexion tests using a simple two-dimensional model. The effect of arm posture on the EMG versus shoulder load moment relationship was studied on the following muscles: supraspinatus, infraspinatus, trapezius (three parts), deltoid (two parts) and pectoralis major. All muscles participated in the MVC tests performed, and its was not possible to suggest a single recommended test for each muscle. Differences in normalized EMG median values ranging up to 30% of MVC were found between linear and curvilinear normalization methods. Short-term repeatability of normalization based on a contraction with gradually increasing force was good. Arm posture affected the relationships between shoulder load moment and EMG activity of all muscles studied. Arm posture did not, however, have a significant effect on the estimated amplitude probability distribution functions during the simulated work task. Therefore, at least for the tasks studied, the principle of normalizing in the middle position of the range of movement was deemed acceptable. PMID- 8223531 TI - Changes in the heart rate and electromyogram beyond the limit time of an isotonic isometric contraction. AB - Nine men [24.6 (SEM 1.1) years] carried out isometric contractions (IC) of the right elbow flexors at 50% and 100% of the maximal voluntary contraction (MVC). At 50% MVC they had to maintain IC until the limit time (isotonic IC: IIC50) and beyond for as long as possible (anisotonic IC: AIC50). At 100% MVC, IC was anisotonic since the decrease in force was immediate (AIC100). Measurements of the force, the integrated electromyogram (iEMG) and the heart rate (fc) were made during the entire period of contraction. There was a linear relationship between the iEMG increase and the fc increase for IIC50 and AIC100. This relationship was not found for AIC50. The role played by the peripheral information would seem to have become more important in fc regulation when the isotonic IC preceding the anisotonic IC was sufficiently long (submaximal IIC). It would seem that the idea of muscle exhaustion at the limit time was only relative, and depended greatly on the subject's motivation and his capacity to endure a certain degree of pain. PMID- 8223532 TI - The effect of short and long duration exercise on serum erythropoietin concentrations. AB - The effects of short and long duration exercise on serum erythropoietin concentrations [EPO]s were studied in seven male cross-country skiers of national team standard and eight male marathon runners, respectively. The short duration exercise was performed as 60 min of cycling at an intensity of 80%-95% of maximal heart rate. Arterial blood oxygen saturations monitored by pulse-oximetry remained unchanged throughout exercise. The partial pressure of O2 at which haemoglobin was half-saturated with O2 calculated from forearm venous blood gas tension and blood O2 saturation, and the erythrocyte 2,3-diphosphoglycerate did not change significantly during the exercise. Blood lactate concentrations were increased at the end of exercise [from 1.3 (SEM 0.1) to 3.6 (SEM 0.3) mmol.l-1]. The [EPO]s determined (by enzyme-linked immunosorbent assay) pre-exercise, 5 min, 6 h, 19 h, and 30 h after the exercise were unchanged [from 16.1 (SEM 2.6) to 19.1 (SEM 3.2), 17.9 (SEM 3.0), 17.0 (SEM 2.5), and 18.6 (SEM 2.9) U.l-1, respectively]. The [EPO]s were not correlated to the earlier parameters. The long duration exercise consisted of habitual training, a 3 week break from training followed by 2 and 4 weeks of re-training. The [EPO]s, body fat (BF), and serum free-testosterone concentrations determined at the end of each period remained unchanged. The maximal oxygen uptakes were decreased after the break from training and increased during retraining (P = 0.04). Body mass (mb) increased after the break in training (P = 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223533 TI - Muscle activation and the isokinetic torque-velocity relationship of the human triceps surae. AB - The influence of muscle activation and the time allowed for torque generation on the angle-specific torque-velocity relationship of the triceps surae was studied during plantar flexion using supramaximal electrical stimulation and a release technique on six male subjects [mean (SD) age 25 (4) years]. Torque-velocity data were obtained under different levels of constant muscle activation by varying the stimulus frequency and the time allowed for isometric torque generation prior to release and isokinetic shortening. To eliminate the effects of the frequency response on absolute torque the isokinetic data were normalized to the maximum isometric torque values at 0.44 rad. There were no significant differences in the normalized torques generated at any angular velocity using stimulus frequencies of 20, 50 or 80 Hz. When the muscle was stimulated at 50 Hz the torques obtained after a 400 ms and 1 s pre-release isometric contraction did not differ significantly. However, with no pre-release contraction significantly less torque was generated at all angular velocities beyond 1.05 rad.s-1 when compared with either the 200, 400 ms or 1 s condition. With a 200 ms pre-release contraction significantly less torque was generated at angular velocities beyond 1.05 rad.s-1 when compared with the 400 ms or 1 s conditions. It would seem that the major factor governing the shape of the torque-velocity curve at a constant level of muscle activation is the time allowed for torque generation. PMID- 8223534 TI - Response of left ventricular diastolic filling to graded exercise relative to the lactate threshold. AB - During incremental exercise, the left ventricular ejection fraction increases up to the intensity of the anaerobic threshold and tends to level off at higher exercise intensities. Since there is a correlation between the response of peak filling rate and ejection fraction to exercise, this study was conducted to determine whether the response of left ventricular diastolic function is similar to the response of systolic function relative to lactate threshold. Twelve healthy men performed two exercise tests on a cycle ergometer. In the first test, lactate threshold and maximal power output were determined. In the second exercise test, gated radionuclide ventriculography was performed at rest, at the lactate threshold intensity, and at peak exercise to measure ejection fraction and peak filling rate. Ejection fraction increased significantly from rest [mean (SD): 62 (5)%] to lactate threshold [76 (7)%] and did not change significantly from lactate threshold to peak exercise [77 (7)%]. Likewise, peak filling rate (normalized for stroke counts) increased from resting [6.1 (0.9) VS.s-1] to lactate threshold [9.4 (1.8) VS.s-1] and did not change significantly from lactate threshold to peak exercise [9.6 (2.9) VS.s-1]. There was no correlation between the change in peak filling rate and the change in ejection fraction from rest to lactate threshold. Thus, during incremental exercise, left ventricular diastolic function responds qualitatively similar to systolic function. PMID- 8223535 TI - Effects of exercise cessation on lipids and lipoproteins in distance runners and power athletes. AB - The purpose of this study was to determine the effects of short-term exercise cessation on lipid and lipoprotein profile and insulin sensitivity in highly trained runners (n = 12; mean age 19.9 years) and power athletes (n = 12; mean age 24.4 years). Following 14 days of exercise cessation, running time to exhaustion and maximal oxygen uptake decreased by 9.2% and 4.8% (P < 0.05) in the runners, while in the power athletes one repetition maximum squat and bench press did not change (P > 0.05). No changes occurred in body composition. Data from a 2 h oral glucose tolerance test revealed an impairment of the glycemic state in all athletes (P < 0.05). In contrast, exercise cessation did not significantly (P > 0.05) alter plasma levels of cholesterol, triglycerides, and low density (LDL) and high density lipoprotein (HDL). No changes were observed in HDL2, HDL2b, and HDL3 subfractions, LDL diameter, and qualitative LDL pattern (P > 0.05). These data thus suggest that despite a decrease in insulin sensitivity, short-term exercise cessation, independent of exercise mode, was insufficient to alter plasma lipid and lipoprotein profiles in well-trained athletes. PMID- 8223536 TI - The influence of tonic neck reflexes on voluntary fatiguing elbow movements in humans. AB - Tonic neck reflexes (TNR) are often assumed to be included in the building of voluntary motor programmes. The present study was designed to test the importance of TNR in the performance by healthy human adults of high level dynamic exercise. The subjects were placed in an experimental situation similar to the original one used by previous authors, but in a fixed restrained posture. They repeatedly lifted weights by extending the elbow, with the head rotated either towards the side of the active upper limb or towards the opposite side. The rate of movement was 0.5 Hz, and the initial amplitude 60 degrees. Successive series of 15 movements separated by an equivalent period of rest were made by each subject until exhaustion. The mechanical work performed during each series of movements was calculated. Surface electromyograms (EMG) from triceps brachii, anconeus, and biceps brachii muscles were recorded simultaneously with the elbow rotation displacement and then integrated. The EMG from the sterno-cleido-mastoidei muscles were also recorded. With successive series of lifting, movement amplitude decreased progressively as a result of fatigue. Thus there was a decrease in the work performed. In agreement with the experimental data of the previous authors as well as with the expected effect of TNR, the decrease in work was less marked when the subjects kept their heads turned towards their moving limb. However, in contrast with the results from other authors, the facilitation was very low. From these results, we concluded that TNR can slightly influence the amount of work that a subject can produce.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223537 TI - Is energy substrate mobilization a limiting factor for cold thermogenesis? AB - Energy substrate mobilization has been suggested as being a limiting factor for the rate of cold-induced thermogenesis (M), and consequently in delaying hypothermia. The evidence supporting this hypothesis in humans, however, is not convincing and the hypothesis has yet to be tested in a rigorous manner using a full heat balance analysis (partitional calorimetry). The goal of this study was therefore to re-investigate whether enhancing energy substrate mobilization by feeding cold-exposed subjects would improve M and affect heat debt (S; the minute by-minute balance of M and heat losses) as well as rectal (Tre) and mean skin temperatures (Tsk). Nine healthy semi-nude fasted subjects were exposed to 5 degrees C (3 h at rest, 1 m.s-1 wind) on three occasions following the ingestion at min 0 and 90 of either: (1) a placebo, (2) 710 kJ of pure carbohydrates (100% CHO), or (3) 710 kJ of a high-carbohydrate bar (High-CHO). As expected in the cold, Tre and Tsk decreased whereas M, S and heat losses increased (P < 0.01). However, there were no differences between treatments, including the final Tre [mean (SEM); 36.4 (0.2); 36.5 (0.3) and 36.5 (0.2) degrees C for the placebo, 100%-CHO and High-CHO tests, respectively]. During the 100%-CHO treatment, rates of carbohydrate oxidation were the highest and fat oxidation the lowest (P < 0.05), whereas the High-CHO treatment caused smaller changes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223538 TI - Effect of graded changes in extracellular muscle volume on cardiovascular drives during static exercise. AB - The effects of graded changes in peripheral extracellular volume on heart rate and blood pressure during isometric exercise were studied in 12 healthy male subjects. Each subject performed four calf ergometer tests with each calf. In all tests, static plantar flexion of one foot was performed in a supine body position with the knee joint flexed to 90 degrees. After a pre-exercise period of 18 min, during which the calf volume was manipulated, the subjects had to counteract a spring force of 120 N for 8 min. In the pre-exercise period the peripheral extracellular volume of the calf muscle to be tested was manipulated in four ways. Test 1:15 min of rest in the exercise position. During the last 3.5 min, the calf volume was increased by venous congestion [80 mmHg (10.67 kPa) applied to the distal part of the thigh by pneumatic cuff]. Test 2: the same protocol as in test 1 but with 7.5-min venous congestion. Test 3:15 min of venous congestion. Test 4: the calf volume was decreased by a negative hydrostatic pressure for 15 min (calf raised about 40 cm above heart level). To clamp the changed calf volume, the thigh cuff was rapidly inflated to 300 mmHg (40.0 kPa) at the end of the volume manipulation and the subjects remained resting for a further 3 min. In test 4, the leg of the subject was passively brought into the exercise position. The occlusion was maintained until 2 min after exercise. The calf volume manipulation led to changes ranging from +105 ml (test 3) to -134 ml (test 4) as measured by water displacement plethysmography. The blood pressure response to exercise was inversely related to the calf volume changes while the heart rate response during exercise showed no clearcut relationship to the pretreatments. PMID- 8223539 TI - A model of sporting performance constructed from autonomic nervous system responses. AB - Four autonomic nervous system (ANS) variables: skin potential and resistance, skin blood flow and temperature - plus instantaneous heart rate and respiratory frequency - were simultaneously recorded during sporting competitions. The performances of 15 marksmen and 7 archers were analysed by comparing the variations in the six parameters during a concentration phase prior to shooting and during the shooting (or active) phase proper. The results of the performances distinguished two groups of sportsmen: firstly, the best shots and secondly, the worst shots. Comparison of the autonomic responses between the groups showed statistically significant correlations among the ANS variables. This made possible a relationship between performance characteristics and a percepto effector factor to be defined. More precisely, this factor was defined as the number of correlations of the measured variables in the two phases, concentration and shooting. A positive relationship was demonstrated--the higher the factor, the better the performance. In addition, the stability of the factor was greater within the best marksmen and archers group. A model of the temporal change of the percepto-effector factor in both groups was given in three dimensions. This modelling of performance characteristics gave a visual indication of the differences between best/worst performances. PMID- 8223540 TI - The influence of air temperature on the EMG/force relationship of the quadriceps. AB - Surface electromyography (EMG) in the past has been used to estimate the intensity of muscle contraction. These estimates were derived from the EMG/force relationship measured at room temperature. How the surface EMG signal is influenced by varying air temperature as it relates to the EMG/force relationship has yet to be investigated. Thus, this study evaluated the influence of different air temperatures (10, 23 and 40 degrees C) on surface EMG during contractions of the quadriceps muscle. Ten subjects [mean (SD) age 29 (7) years and weight 78.3 (7.8) kg] performed a criterion task of five contractions ranging from 10 to 100% of a maximal voluntary contraction, five times over a 1.5-h period in all conditions. The EMG signals generated from the rectus femoris, and the forces associated with the contractions, were captured on FM tape and subsequently digitized at a sampling rate of 2000 Hz. The relationship between EMG and force was different under the different conditions; EMG was reduced at a given force as temperature increased, and the EMG increased in the 10 degrees C environment over time. The differences that occurred at the various temperatures were believed to be related to fluid distribution in the muscle, muscle conduction velocity and sweating. The data imply that the EMG/force relationship measured using surface electrodes is influenced by ambient temperature. PMID- 8223541 TI - Metabolic consequences of repeated exercise in long distance runners. AB - To assess the rates of change in muscle metabolites such as phosphocreatine (PCr) and inorganic phosphate (Pi) during repeated exercise sessions with rest periods, 31-phosphorus nuclear magnetic resonance spectroscopy was used for continuous and noninvasive measurements. Five long-distance runners and six healthy male subjects as controls performed a 2-min femoral flexion exercise at 20 kg.m.min-1 in a 2.1 T superconducting magnet with a 67-cm bore; they repeated this exercise four times with a 2-min rest period. At the beginning of exercise, PCr decreased exponentially; at the end, it increased. During exercise and in the early phase of the recovery in every exercise session, the PCr values were significantly higher in the long-distance runners than in the control subjects (P < 0.05). The Pi increases and decreases involved with exercise also revealed exponential changes. The Pi values did not significantly differ during exercise; however, Pi recovery was faster in the long-distance runners than in the control subjects (P < 0.05). The Pi:PCr ratio during exercise increased linearly with exercise; and Pi:PCr during recovery was smaller in the long-distance runners than in the control subjects (P < 0.05). In conclusion, the long-distance runners revealed faster PCr and Pi kinetics after exercise and a smaller Pi:PCr during exercise than did the control subjects. It is suggested that these results were attributable to a greater oxidative capacity of muscles in the long-distance runners. PMID- 8223542 TI - The influence of electrode position on bipolar surface electromyogram recordings of the upper trapezius muscle. AB - The effect of electrode position on the upper trapezius muscle on the myo electric signal amplitude was investigated with special reference to arm position and estimate of force output. Previously, a depression of the electromyogram (EMG) signal has been reported midway between the seventh cervical vertebrae (C7) and acromion (Veiersted 1991, Eur J Appl Physiol 62:91-98) although this electrode position has been recommended (Zipp 1982, Eur J Appl Physiol 50: 41 54). Ten healthy subjects performed maximal shoulder elevations with the arm in vertical, abducted and flexed positions and they performed a dynamic movement test. The myo-electric signal was recorded along the length of the right upper trapezius muscle by a 16-channel bipolar array electrode and was integrated with a 0.2-s time resolution. A region just lateral to the midpoint between C7 and the lateral edge of acromion was found with high and stable amplitudes (% coefficient of variation equalled 5.6). At the midpoint a dip in the amplitude profile appeared which was slightly displaced by arm abduction or flexion probably due to sliding of the skin relative to the muscle. A linear EMG-force relationship was found in the region with high signal amplitudes, whereas the more lateral and the dip region showed highly variable EMG-force relationships. Thus, it was found that when using bipolar surface electrodes with an interelectrode distance of 2 cm a centre position 2 cm lateral to the midpoint between C7 and acromion provided good repeatability and high signal yield. PMID- 8223543 TI - Changes in intracellular pH during repeated exercise. AB - The rates of change in intracellular pH during repeated exercise sessions with rest periods was determined by 31 phosphorus-nuclear magnetic resonance spectroscopy (31P-MRS). Five long-distance runners and six healthy male subjects as controls performed a 2-min femoral flexion at 20 kg.m.min-1 in a 2.1 T superconducting magnet with a 67-cm bore and repeated this exercise four times with 2-min rest periods intervening. In all cases during exercise the inorganic phosphate (Pi) peak split into two, the earlier increased rapidly (high-pH Pi) and the later (low-pH Pi) increased more slowly. The Pi peaks were separated by a fitting procedure using the least square mean method. The high-pH Pi area during exercise decreased as the number of repeated exercise periods increased, while the low-pH Pi area gradually increased. Although the total Pi area decreased exponentially during the recovery period, the high-pH Pi area decreased first and then the low-pH Pi area reduced gradually. The pH values were estimated from the chemical shift between the phosphocreatine peak and each split peak in the Pi. The high-pH in pooled data ranged from 6.6 to 7.0 during exercise and recovery, while the low pH decreased to 6.2 during exercise. As the number of exercise periods increased, each pH value gradually became less acidic, although there was a tendency to more acidity in the control subjects than in the long-distance runners.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223544 TI - Influence of the menstrual cycle and oral contraceptives on thermoregulatory responses to exercise in young women. AB - Thermoregulatory responses to exercise in relation to the phase of the menstrual cycle were studied in ten women taking oral contraceptives (P) and in ten women not taking oral contraceptives (NP). Each subject was tested for maximal aerobic capacity (VO2max) and for 50% VO2max exercise in the follicular (F) and luteal (L) phases of the menstrual cycle. Since the oral contraceptives would have prevented ovulation a quasi-follicular phase (q-F) and a quasi-luteal phase (q-L) of the menstrual cycle were assumed for P subjects. Exercise was performed on a cycle ergometer at an ambient temperature of 24 degrees C and relative air humidity of 50%. Rectal (Tre), mean skin (Tsk), mean body (Tb) temperatures and heart rate (fc) were measured. Sweat rate was estimated by the continuous measurement of relative humidity of air in a ventilated capsule placed on the chest, converted to absolute pressure (PH2Ochest). Gain for sweating was calculated as a ratio of increase in PH2Ochest to the appropriate increase in Tre for the whole period of sweating (G) and for unsteady-state (Gu) separately. The VO2max did not differ either between the groups of subjects or between the phases of the menstrual cycle. In P, rectal temperature threshold for sweating (Tre,td) was 37.85 degrees C in q-L and 37.60 degrees C in q-F (P < 0.01) and corresponded to a significant difference from Tre at rest. The Tre, Tsk, Tb and fc increased similarly during exercise in q-F and q-L. No menstrual phase-related differences were observed either in the dynamics of sweating or in G.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223545 TI - Plasma atrial natriuretic factor during cold-induced diuresis. AB - The purpose of this study was to evaluate the possible contribution of atrial natriuretic factor (ANF) to cold-induced diuresis. Seven healthy men, dressed in shorts, were exposed to a cold environment (+12 degrees C) for 90 min, and also to a thermoneutral environment. Exposure to cold increased urine output and sodium excretion significantly but plasma ANF concentration remained unchanged. The increase in urinary potassium excretion during cold exposure was not significant (P = 0.0636) and plasma renin activity did not change either. Exposure to cold increased mean arterial pressure significantly but it did not affect heart rate. We concluded that acute exposure to the cold environment induced a diuretic response, which was a solute diuresis in its nature. Our results did not give support to the hypothesis that ANF might be involved in the renal response to cold exposure. PMID- 8223546 TI - Structural study of a cell-wall mannan of Saccharomyces kluyveri IFO 1685 strain. Presence of a branched side chain and beta-1,2-linkage. AB - Acetolysis of the cell-wall mannan of Saccharomyces kluyveri under mild conditions, gave fragments with 1-6 mannose residues. The structures of mannopentaose and mannohexaose were determined to be [Formula; see text] respectively, by two-dimensional homonuclear Hartmann-Hahn spectroscopy and a sequential NMR assignment method that combines 1H-13C correlated spectroscopy, relayed coherence transfer spectroscopy, 1H-detected heteronuclear multiple-bond connectivity and methylation analysis. The H1 proton chemical shift of a neighboring alpha-1,2-linked mannose unit of the 3-O-substituted structure was shifted upfield by the addition of a mannose unit to the adjacent 3-O-substituted unit by an alpha-1,6 linkage. The characteristic H1--H2-correlated cross-peak of the alpha-1,3-linked mannose unit substituted by a beta-1,2 linkage, beta 1- >2Man alpha 1-->3, in the mannan of S. kluyveri, as also found by two-dimensional homonuclear Hartmann-Hahn spectroscopy in the mannan of Candida guilliermondii, a pathogenic yeast in man. PMID- 8223547 TI - Cloning, DNA sequence analysis and partial characterization of pepN, a lysyl aminopeptidase from Lactobacillus delbruckii ssp. lactis DSM7290. AB - In cell extracts of Lactobacillus delbruckii ssp. lactis DSM7290 a peptidase with the ability to hydrolyse Phe-beta-naphthylamide (Phe-beta-NA) and His-beta-NA could be detected. Escherichia coli lacking the enzyme activity in an enzymic plate assay was used to screen high-copy-number and low-copy-number plasmid libraries of size-fractionated Lactobacillus DNA. Clones with the desired phenotype were detected, and the gene, designated pepN, was further subcloned and sequenced. A large open reading frame of 2529 nucleotides is predicted to encode a protein of 843 amino acids (95358 Da). Comparison of the pepN gene from Lb. delbruckii ssp. lactis DSM7290 indicates that it is homologous to genes of the family of Zn(2+)-metallohydrolases and PepN shows identity with the active centre Zn(2+)-binding motif of these enzymes. The substrate Lys-beta-NA is more effectively cleaved than Phe-beta-NA or His-beta-NA which were used for screening in E. coli. The cloned pepN gene was efficiently overexpressed in E. coli and subcloning of the gene in Lactobacillus casei resulted in a moderate overexpression of approximately 20-fold. The pepN gene product was purified from the pepN-deficient E. coli strain CM89, using the substrate Lys-p-nitroanilide (Lys-NH-Ph) in the assay procedure. In a four-step procedure including streptomycin sulfate precipitation, anion-exchange chromatography and gel filtration the peptidase was purified to electrophoretic homogeneity. PMID- 8223548 TI - Cloning, sequencing and immunological characterization of the corrinoid containing subunit of the N5-methyltetrahydromethanopterin: coenzyme-M methyltransferase from Methanobacterium thermoautotrophicum. AB - A 3.5-kb EcoRI fragment of the Methanobacterium thermoautotrophicum chromosome contains five open reading frames, mtrA to mtrE. The deduced N-terminal amino acid sequence of mtrA is identical with 26 N-terminal amino acids of a corrinoid containing membrane protein from Methanobacterium. Computer-aided analyses of mtrA predicts 237 amino acids with a molecular mass of 25,603 Da for its gene product. A hydropathy plot of this amino acid sequence indicates one hydrophobic helical conformation near the N-terminus of the peptide which represents a tentative membrane-spanning region. The main part of the protein, however, shows hydrophilic domains, suggesting a location outside the cytoplasmic membrane. These domains are probably accessible by monospecific polyclonal antibodies raised previously against the corrinoid-containing membrane protein. The immunogold-labeling technique revealed that the corrinoid-dependent membrane protein was detectable at the cytoplasmic face of the membranes and of vesicle preparations. No significant identity of the deduced amino acid sequence was found with sequences of several corrinoid-containing enzymes. In contrast to the hydrophilic gene product of mtrA, four other gene products from the gene cluster encode extremely hydrophobic proteins. The N-terminal sequences of mtrC and mtrD are identical with two peptides of the N5-methyltetrahydromethanopterin:coenzyme M methyltransferase complex from Methanobacterium, indicating that the mtr genes encode this membrane protein. PMID- 8223549 TI - The mannose-specific lectins from ramsons (Allium ursinum L.) are encoded by three sets of genes. AB - Lectin cDNA clones encoding the two mannose-binding lectins from ramsons (allium ursinum L.) bulbs, AUAI and AUAII (AUA, Allium ursinum agglutinin), were isolated and characterized. Sequence comparison of the different cDNA clones isolated revealed three types of lectin clones called LECAUAG0, LECAUAG1 and LECAUAG2, which besides the obvious differences in their sequences also differ from each other in the number of potential glycosylation sites within the C-terminal peptide of the lectin precursor. In vivo biosynthesis studies of the ramson lectins have shown that glycosylated lectin precursors occur in the organelle fraction of radioactively labeled ramson bulbs. Despite the similarities between the A. ursinum and the A. sativum (garlic) lectins at the protein level, molecular cloning of the two ramson lectins has shown that the lectin genes in A. ursinum are organized differently. Whereas in A. sativum the lectin polypeptides of the heterodimeric ASAI are encoded by one large precursor, those of the heterodimeric AUAI lectin are derived from two different precursors. These results are confirmed by Northern blot hybridization of A. ursinum RNA which, after hybridization with a labeled lectin cDNA, reveals only one band of 800 nucleotides in contrast to A. sativum RNA which yields two bands of 1400 and 800 nucleotides. Furthermore it is shown that the two mannose-binding lectins are differentially expressed. PMID- 8223550 TI - Nuclear magnetic resonance study of the codon-anticodon interaction in Bombyx mori tRNA(GCCGly). AB - NMR spectra of Bombyx mori tRNA(GCCGly) were recorded in the GCC absence and presence of the oligonucleotide, GGCUp, which contains the codon sequence, GGC. The difference between the spectra with and without the codon oligonucleotide indicates the appearance of five new imino proton peaks. For the assignment of these peaks, G*GCUp, in which the 5'-terminal G was enriched with 95% 15N, was prepared (G*, 15N-labeled guanosine). In the imino proton spectrum of B. mori tRNA(GCCGly) on the addition of G*GCUp, the peak at 12 ppm became a doublet due to coupling with 15N nuclei. In the two-dimensional 1H-15N heteronuclear multiple quantum correlation (HMQC) spectrum, only the peak at 12 ppm was observed, and thus it was assigned to the imino proton of the 5'-terminal G of GGCUp interacting with tRNA(GCCGly). Judging from the temperature effect and chemical shifts, the five new imino proton peaks are presumed to be due to three G.C base pairs, induced by the codon-anticodon interaction, and one U.U base pair, induced by an interaction between the 3' terminal U of GGCUp and U33 neighboring the anticodon. The binding of three trinucleotides (GGCp, GGUp and GCUp) to B. mori tRNA(GCCGly) was also investigated. Ultracentrifugation analysis showed that tRNA(GCCGly) underwent dimerization through the anticodon-anticodon interaction, but the dimerization was broken on addition of GGCUp. On 1H-NMR and ultracentrifugation analysis, it was found that GCUp not complementary to the anticodon also binds to the anticodon loop. PMID- 8223551 TI - Activation of porcine pepsinogen A. The stability of two non-covalent activation intermediates at pH 8.5. AB - Reaction products formed during activation of porcine pepsinogen A at pH 2 were characterized by native agar-gel electrophoresis and by denaturing SDS/PAGE. The results revealed the presence of non-covalent intermediates between prosegment peptides and pepsin. The complexes Leu1p-Leu44p/pepsin and Leu1p-Leu16p/pepsin were isolated (the prosegment residues are characterized by the suffix p; numbering of residues starts again from the N-terminus of pepsin). Relative to mature pepsin, the inherent milk-clotting activities of the intermediates were 3% and 18%, respectively. The intermediates were incubated at pH 8.5 for 20 min at 28 degrees C and the residual proteolytic activities were tested at pH 2. The stabilities at pH 8.5 were between those of pepsinogen and pepsin, Leu1p Leu44p/pepsin being most stable. The implications of these findings for the conformational changes that occur during the activation of pepsinogen are discussed. PMID- 8223552 TI - Further characterization of structural and electric properties of non-spherical alpha-crystallin. AB - Recently we have shown that the population of native alpha-crystallin, isolated using size-exclusion chromatography from eye lenses of calves, is multimodal. Most of the protein probably possesses an almost spherical appearance, but at least one of the other modes represents more extended, ellipsoidally and/or cylindrically shaped molecules [Van Haeringen, B., Eden, D., Van den Bogaerde, M.R., Van Grondelle, R. & Bloemendal, M. (1992) Eur. J. Biochem. 210, 211-216]. In the present study, we characterize various subpools of a single alpha crystallin size-exclusion chromatography elution peak by means of transient electric-birefringence measurements, ultraviolet linear-dichroism spectroscopy and analytical fast protein liquid chromatography. It is concluded that the fractions have a well-defined stable mass and are not in reversible equilibrium with each other. All pools appear to be composed of at least two types of differently shaped molecules. The hydrodynamic dimensions and electric properties of the different alpha-crystallin species are characterized. The non-spherical alpha-crystallin is found to be optically and electrically more anisotropic, and to contain a larger permanent electric dipole moment than the spherical form. A model for the composition of the alpha-crystallin pool is presented. PMID- 8223553 TI - Amino acid sequence of the small cadmium-binding protein (MP II) from Nereis diversicolor (annelida, polychaeta). Evidence for a myohemerythrin structure. AB - The primary sequence of the low-molecular-mass cadmium-binding protein metalloprotein II of Nereis diversicolor (Hediste diversicolor, recent denomination) has been determined. This protein is composed of 119 amino acids and has 80.8% identity with the N. diversicolor myohemerythrin [Takagi, T. & Cox, J. A. (1991) FEBS Lett. 285, 25-27]. The fact that iron, which normally binds to myohemerythrin, is not found to be associated with the cadmium-binding protein metalloprotein II in cadmium-exposed animals could be the result of the complete abolition of the iron-binding capacity of the protein due to the binding of cadmium. PMID- 8223554 TI - Cloning, site-specific mutagenesis, expression and characterization of full length chloroplast NADP-malate dehydrogenase from Pisum sativum. AB - Chloroplast NADP-dependent malate dehydrogenase is regulated by a dithiol redox reaction. The assignment of the groups involved, requires the primary structure of the enzyme to be known. Using the polymerase chain reaction and the cDNA library of Pisum sativum, the sequence of the enzyme and its targeting signal was determined. The gene was cloned in Escherichia coli JM83 and expressed in E. coli JM83 and E. coli B at high yield. The determination of the physical properties of the gene product proves the recombinant protein to be indistinguishable from the enzyme purified from the plant. This holds true, in spite of the fact that the plant enzyme lacks 11 N-terminal residues. The lengths of the complete polypeptide chain of the recombinant enzyme and its transit peptide are 388 and 53 residues, respectively. The comparison of the sequences of the mature enzyme with those of known chloroplast NADP-MDH shows 83-95% identity, but with mitochondrial or bacterial MDH only approximately 20%. Reduction of the (inactive) oxidized enzyme with dithiothreitol allows mimicking of the in vivo activation. The reaction follows a consecutive second-order-kinetics mechanism. Guanidinium chloride (GdmCl) at concentrations below 0.4 M leads to a significant activation of the oxidized form of the enzyme. At [GdmCl] = 0.4-0.46 M, both oxidized and reduced NADP-MDH show highly cooperative changes in the hydrodynamic and spectral properties, indicating the synchronous breakdown of the quaternary, tertiary and secondary structures. Site-directed mutations C23A and C28A do not quench the regulatory properties of the enzyme; additional substitution of alanine for Cys206 and Cys376 renders the enzyme equally active in both the reduced and the oxidized state. Therefore, one can consider these residues, either alone or in combination with Cys23 and Cys28, as responsible for enzyme activation. PMID- 8223555 TI - Heterogeneity of glycosylphosphatidylinositol-anchored alkaline phosphatase of calf intestine. AB - A method is described for large-scale purification of glycosylphosphatidylinositol-anchored alkaline phosphatase from intestinal mucosa and chyme to homogeneity. Both enzyme preparations contain approximately 2 mol fatty acid/mol subunit and exhibit a very similar fatty acid composition with octadecanoate and hexadecanoate as prevalent components. No significant differences between native glycosylPtdIns-anchored and hydrophilic alkaline phosphatases from both sources were found regarding Km, Vmax, the type of inhibition and inhibition constants of the amino acids L-leucine, L phenylalanine, and L-tryptophan. The purified enzymes of both sources yield diacylglycerol and phosphatidic acid, after treatment with phosphatidylinositol specific phospholipase C (PtdIns-PLC) and glycosylphosphatidylinositol phospholipase D (PLD), respectively. Enzyme preparations of both sources appear as heterogeneous mixtures of five fractions separable by octyl-Sepharose chromatography. Fraction I corresponds to the anchorless enzyme, fractions II-V differ in their susceptibility to phospholipases. Fractions II and IV are completely split by PtdIns-PLC or PLD action, almost 50% of fraction III is split by PtdIns-PLC, while fraction V is resistant. The susceptibility of these two fractions toward the action of PLD is considerably higher. Fatty acid analysis yields molar ratios of fatty acids/alkaline phosphatase subunit of 1.78, 2.58, 2.24, and 3.37 for fractions II, III, IV, and V, respectively. Aggregates of glycosylPtdIns-anchored alkaline phosphatase of all fractions are seen in native PAGE in the presence of Triton X-100. By gel chromatography in the presence of Brij 35, fractions II-V form stable multiple aggregates of dimers and may bind different amounts of the detergent. These data, together with fatty acid analysis, can be interpreted by the following model. Fractions II and IV are tetramers and octamers with two molecules fatty acid/subunit. Fraction III is a tetramer, bearing one additional fatty acid molecule, localized on the dimer. Fraction V is an octamer, containing glycosylPtdIns-anchor molecules with three molecules fatty acids/anchor molecule. The additional fatty acid residue is possibly located on inositol and responsible for the reduced susceptibility to PtdIns-PLC. The similarity of all measured parameters of both enzymes suggests that the glycosylPtdIns-anchored alkaline phosphatase of the mucosa is released into the chyme without changing the anchor molecule constituents. PMID- 8223556 TI - Side-chain specificities of human and bovine cytochromes P-450scc. AB - Cytochrome P-450scc catalyses the conversion of cholesterol to pregnenolone by the sequential hydroxylation of the side chain of cholesterol. This occurs at a single active site and produces 22R-hydroxycholesterol and 22R-20 alpha dihydroxycholesterol as intermediates. To further define the active site of human and bovine cytochromes P-450scc, we have examined the kinetics of the conversion of structural analogues of cholesterol with modified side chains, to pregnenolone. Analysis of the side-chain cleavage of analogues of cholesterol modified at C22 confirmed the high degree of structural specificity for the 22R position by cytochrome P-450scc, the major effect being on the turnover number (kcat) rather than on binding. The analogues of cholesterol that had a polar group at C24, C25 or C26 had much lower Km values and generally lower kcat values than the non-polar analogues which were tested. Km values of the polar analogues were 3-25-times lower than the Km for cholesterol and kcat values were also much lower than the kcat values for cholesterol, particularly for the human enzyme. The data suggest that the tight binding of the analogues with a hydroxyl or ketone group at C24, C25 or C26 places C20 and C22 in a poor orientation relative to the heme group for hydroxylation to occur. Many of the polar analogues which were tested are postulated regulators of cellular cholesterol metabolism. Several of these analogues are good substrates for bovine and human cytochromes P-450scc at low substrate concentration, as determined from their kcat/Km values. This study also indicates that the active site of cytochrome P-450scc is well conserved between bovine and human cytochromes. However, small species differences are evident since lower kcat values relative to the kcat of cholesterol are observed for some polar side-chain analogues of cholesterol with the human enzyme. PMID- 8223557 TI - Aberrant proteolysis of the beta-amyloid precursor protein in familial Alzheimer's disease lymphoblastoid cells. AB - Lymphoblastoid cells from patients with early-onset and late-onset familial Alzheimer's disease showed increased expressions of beta-amyloid precursor mRNA and protein as well as interleukin-1 and alpha 1-antichymotrypsin protein. Early onset and late-onset familial Alzheimer's disease cells had greater production of 16-kDa beta-amyloid C-terminal preamyloid peptides than did normal cells. A pulse chase experiment indicated that aberrant processing of this peptide resulted in its abnormal accumulation. Furthermore, the peptide prepared from early-onset familial Alzheimer's disease cells using formic acid could be separated into four discrete fragments. The N-terminal amino acid sequencing of each fragment indicated that the 16-kDa peptide was generated by cleavage, principally at the 30 amino acids N-terminal to beta-amyloid. Both the enhanced synthesis and aberrant processing of the beta-amyloid precursor protein, therefore, are basic processes associated with familial Alzheimer's disease lymphoblastoid cells. PMID- 8223558 TI - Abduction of iron(III) from the soluble methane monooxygenase hydroxylase and reconstitution of the binuclear site with iron and manganese. AB - The apo-form of the soluble methane monooxygenase hydroxylase from Methylococcus capsulatus (Bath) was prepared via chelation of iron(III) with 3,4 dihydroxybenzaldehyde. The apohydroxylase was reconstituted by the anaerobic addition of Fe(II) followed by air oxidation. The enzyme thus prepared regained 85-90% of its original catalytic activity. The incorporation of two manganese(II) ions/mol of apohydroxylase was monitored by EPR spectroscopy. The Mn(II) ions occupy the native diiron active site and remain in the +2 oxidation state. The EPR data suggest strong coupling between the two Mn(II) ions and retention of the mu-hydroxo (alkoxo) bridge. The results of this study indicate that the M. capsulatus (Bath) hydroxylase contains a single diiron site. PMID- 8223559 TI - Localization of a putative magnesium-binding site within the cytoplasmic domain of the sarcoplasmic reticulum Ca(2+)-ATPase. AB - The amino acid sequences of several P-type ATPases share regions of high similarity. The functions of some of these regions, although several proposals have been made, have not yet been absolutely identified. In particular, one of these domains, located within the cytoplasmic loop in the area known as the 'hinge' domain, exhibits the highest degree of conservation. In the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA-1), this region is located at residues 700-712. Comparison of the sequence in this domain with calcium-binding proteins reveals similarities with the center of the helix-loop-helix EF-hand structure that is known to form divalent-cation-binding sites. A 38-residue polypeptide, corresponding to the domain 682-719 of the Ca(2+)-ATPase was synthesized and tested for its ability to bind divalent cations. Circular-dichroism, intrinsic fluorescence and fluorescence-energy-transfer studies performed on this polypeptide in solution support the hypothesis that this domain has, in the protein, the ability to bind a divalent cation, presumably Mg2+, with an affinity of 10-15 mM. This property is observed for the isolated polypeptide in aqueous solvent and in the presence of low concentrations of the alpha-helix promoter 2,2,2-trifluoroethanol. Substitution of either one or two critical amino acids in the sequence induces a significant reduction of the binding properties. It is proposed that this sequence is involved in the co-ordination of a Mg2+ in the nucleotide-binding site and/or in the phosphorylation site of P-type ATPases. PMID- 8223560 TI - Hydrogen formation from glycolate driven by reversed electron transport in membrane vesicles of a syntrophic glycolate-oxidizing bacterium. AB - Oxidation of glycolate to 2 CO2 and 3 H2 (delta G degrees' = +36 kJ/mol glycolate) by the proton-reducing, glycolate-fermenting partner bacterium of a syntrophic coculture (strain FlGlyM) depends on a low hydrogen partial pressure (pH2). The first reaction, glycolate oxidation to glyoxylate (E zero' = -92 mV) with protons as electron acceptors (E zero' = -414 mV), is in equilibrium only at a pH2 of 1 microPa which cannot be maintained by the syntrophic partner bacterium Methanospirillum hungatei; energy therefore needs to be spent to drive this reaction. Glycolate dehydrogenase activity (0.3-0.96 U.mg protein-1) was detected which reduced various artificial electron acceptors such as benzyl viologen, methylene blue, dichloroindophenol, K3[Fe(CN)6], and water-soluble quinones. Fractionation of crude cell extract of the glycolate-fermenting bacterium revealed that glycolate dehydrogenase, hydrogenase, and proton-translocating ATPase were membrane-bound. Menaquinones were found as potential electron carriers. Everted membrane vesicles of the glycolate-fermenting bacterium catalyzed ATP-dependent H2 formation from glycolate (30-307 nmol H2.min-1 x mg protein-1). Protonophores, inhibitors of proton-translocating ATPase, and the quinone analog antimycin A inhibited H2 formation from glycolate, indicating the involvement of proton-motive force to drive the endergonic oxidation of glycolate to glyoxylate with concomitant H2 release. This is the first demonstration of a reversed electron transport in syntrophic interspecies hydrogen transfer. PMID- 8223561 TI - Isolation and characterization of three cDNAs coding for Rab proteins from the albumen gland of the mollusc Lymnaea stagnalis. AB - Three cDNA clones encoding small GTP-binding proteins, LS-Rab1, LS-Rab2 and LS Rab18a were isolated from a cDNA library from the albumen gland of the pulmonate snail Lymnaea stagnalis. Comparison of the deduced amino acid sequences with available sequences from the EMBL/Data Bank revealed that LS-Rab1 and LS-Rab2 show a sequence identity of 89-90% to the mammalian Rab1 and Rab2 proteins, and can therefore be regarded as the L. stagnalis homologs. LS-Rab18a may be considered a new member of the Rab subfamily, closely related to mouse Rab18 (74% amino acid identity). Interestingly, LS-Rab1 and LS-Rab2 share a very high sequence conservation with their mammalian homologs (95-97%) over the first 178 191 N-terminal amino acids, whereas the C-terminal part is almost completely divergent, except for their extreme ends (2-4 amino acids). The implications of these observations for the understanding of Rab-targeting signals are discussed. The LS-rab cDNAs were expressed in COS-7M6 cells. The resulting 22-kDa products were shown to bind GTP. In the albumen gland mRNA, levels of LS-rab1 appeared to be much higher than those of LS-rab2 and LS-rab18a, suggesting an important role for the LS-Rab1 protein in the albumen gland. PMID- 8223562 TI - Intracellular and extracellular processing of chromogranin A. Determination of cleavage sites. AB - Chromogranins are a family of acidic soluble proteins which exhibit widespread distribution in endocrine cells and neurons. Chromogranin A (CGA), the major soluble component of the secretory granules in chromaffin cells of the adrenal medulla, is a single polypeptide chain of 431 residues with an apparent molecular mass of 70-75 kDa and a pI of 4.5-5. In mature bovine chromaffin granules about 50% of the CGA has been processed. In the present paper, the structural features of the proteolytic degradation mechanism have been characterized with regard to the possible function of CGA as a prohormone, as suggested by recent studies. CGA derived components present in chromaffin granules were subjected to either two dimensional gel electrophoresis or HPLC and the N-terminal of each fragment was sequenced. Immunoblotting with antisera to specific sequences within the CGA molecule were used to characterize these fragments further at their C-terminal. In addition, a similar approach was performed to characterize CGA-derived fragments released into the extracellular space from directly depolarized bovine cultured chromaffin cells. Our results identified several proteolytic cleavage sites involved in CGA degradation. Intragranular processing occurs at 12 cleavage sites along the peptide chain located in both N- and C-terminal moieties of the protein; a preferential proteolytic attack in the C-terminal part was noted. We found that CGA processing also occurs in the extracellular space after release, generating new shorter fragments. The proteolytic cleavage sites identified in this study were compared with the cleavage points which are thought to be involved in generating CGA fragments with specific biological activity: pancreastatin, chromostatin and N-terminal vasostatin fragments. In addition, a new 12-amino-acid CGA-derived peptide corresponding to the sequence 65-76 was identified in the soluble core of purified chromaffin granules. This short peptide was released, together with catecholamines, after stimulation of cultured chromaffin cells suggesting its presence within the storage complex of chromaffin granules. The specific biological activity of this CGA-derived fragment remains to be determined. PMID- 8223563 TI - Regulation of neurosecretory habituation by cAMP. Role of adaptation of cAMP signals. AB - The role of adaptation in receptors was studied with regard to the processing of neural information by cAMP in PC12 cells. Both secretin and adenosine A2 receptors elevated cAMP levels. During prolonged stimulation, secretin receptors demonstrated adaptation of cAMP levels, while adenosine A2 receptors did not. Incubation of cells with secretin increased the norepinephrine secretion resulting from stimulation by acetylcholine, but not from high potassium depolarization. The difference in the adaptation of cAMP by secretin and adenosine A2 receptors was responsible for the changes observed in norepinephrine secretion. This difference in secretory responses was caused by protein-kinase-C activation, which occurred during acetylcholine stimulation but not during depolarization by high potassium. Secretin receptors also demonstrated a reduced ability to elevate cAMP levels during repetitive stimulation, even for stimulation times where no adaptation of cAMP signals was observed, suggesting that secretin receptors possess the ability to habituate in addition to undergoing adaptation. These results are discussed with regard to the modulatory roles cAMP may play in neurosecretory habituation. PMID- 8223564 TI - Combined plasma-desorption mass spectrometry and Edman degradation applied to simultaneous sequence determination of isoforms of structural proteins from the cuticle of Locusta migratoria. AB - The primary structures of two basic low-molecular-mass proteins, Lm-67 and Lm-70 from the pharate cuticle of the migratory locust, Locusta migratoria, were determined. The sequencing strategy was based on combined use of plasma- desorption mass spectrometry (PDMS) and automatic Edman degradation of the proteins and their enzymically derived peptides. The mass-spectral data showed the presence of two proteins in each preparation. For protein preparation Lm-67, this was indicated by the mass spectrum of the intact protein. For protein preparation Lm-70, the presence of two variants only became evident by mass spectrometric analysis of the enzymically derived peptides. Both proteins show strong similarity to other exocuticular proteins from L. migratoria. PMID- 8223565 TI - Probing the role of metal ions in the mechanism of inositol monophosphatase by site-directed mutagenesis. AB - Since inhibition of myo-inositol monophosphatase (EC 3.1.3.25) by lithium ions and the resulting attenuation of phosphatidylinositol cycle activity may be the mechanism by which lithium exerts its therapeutic effect in the treatment of manic depression, it is of great interest to understand the mechanism of the enzyme and how lithium and other metals interact with it. Divalent magnesium is essential for enzyme activity, whereas Li+ and high concentrations of Mg2+ act as uncompetitive inhibitors with respect to substrate. From the recently solved crystal structure of the human enzyme, several amino acid residues in the active site were targeted for mutagenesis studies. Nine single-residue substituted mutants were characterized with regard to catalytic parameters, Mg2+ dependence, and Li+ inhibition. In addition, a terbium fluorescence assay was developed to determine the metal binding properties of the wild-type and mutant enzymes. Although none of these mutations affected Km for substrate substantially, the mutations Glu70-->Gln, Glu70-->Asp, Asp90-->Asn and Thr95-->Ala, in which residues within coordinating distance of the active site metal were modified, all resulted in large reductions in catalytic activity. The position of Glu70 in the crystal structure further suggests that this residue may be involved in activating water for nucleophilic attack on the substrate. The mutations Lys36- >Ile, Asp90-->Asn, Thr95-->Ala, Thr95-->Ser, His217-->Gln, and Cys218-->Ala all resulted in parallel reductions in both lithium and magnesium affinity, suggesting that Li+ and Mg2+ share a common binding site. PMID- 8223566 TI - Complement fragment C3a stimulates Ca2+ influx in neutrophils via a pertussis toxin-sensitive G protein. AB - The signal pathways of neutrophils following stimulation with the complement fragment C3a (C3a) were studied in neutrophils and compared to the pathways activated by complement fragment C5a (C5a). Analysis of polyphosphoinositol lipid turnover showed that C5a, but not C3a, activated phosphatidylinositol bisphosphate-3-kinase (PtdInsP2 3-kinase) indicating that different signal pathways are activated by the two anaphylatoxins. To examine whether C3a stimulated Ca2+ transients, cytosolic free Ca2+ levels were analyzed in Fluo-3 labelled neutrophils by flow cytometry. C3a stimulated a fast and concentration dependent increase of cytosolic free Ca2+. Comparison of the C3a response with that of C5a revealed a more pronounced C5a-triggered Ca2+ rise. Addition of EGTA to the extracellular buffer prior to stimulation did not significantly alter the initial Ca2+ rise at low C5a concentrations, but reduced the time course of the Ca2+ transients at high concentrations. In marked contrast, EGTA completely blocked the Ca2+ response stimulated by C3a in neutrophils labeled with either Indo-1/AM or Fluo-3. Preincubation of neutrophils with pertussis toxin inhibited both C3a- and C5a-stimulated Ca2+ transients, indicating the involvement of guanine-nucleotide-binding proteins (G proteins) in these processes. In order to examine whether the C3a receptor is coupled to G proteins, binding of guanosine 5'-O-(3-[35S]thiotriphosphate) ([35S]GTP[S]) to purified neutrophil plasma membranes was studied. Both C3a and C5a stimulated high-affinity binding of [35S]GTP[S] up to 1.5-fold and 3-fold, respectively. These data suggest that the two anaphylatoxins activate pertussis-toxin-sensitive G proteins, which then trigger different signal transduction pathways. C3a specifically stimulated Ca2+ influx from the extracellular medium, whereas C5a additionally activated the PtdInsP2 3-kinase and stimulated Ca2+ mobilization from intracellular stores. PMID- 8223567 TI - Identification of important bases in a single-stranded region (SSrC) of the hepatitis delta (delta) virus ribozyme. AB - Models for the secondary structure of genomic and antigenomic self-cleaving RNAs of human hepatitis delta (delta) virus (HDV) have been proposed by several groups. Our recent results support a pseudoknot structure and have allowed us to identify functionally important nucleotides in single-stranded regions [nucleotides 726-731 (SSrA) and nucleotides 762-766 (SSrB)]. For the identification of the important residues in the remaining single-stranded region, nucleotides 708-715 (SSrC), of the genomic HDV ribozyme, we made derivatives with a single-base substitution in the SSrC region. To screen inactive mutants rapidly, we use a simplified in-vitro selection method. Among the various base substitutions in mutants in the SSrC, U708A, C709(A/G/U) and G713C variants had less than 10% of the cleavage activity of the wild-type SSrC (HDV86). By analyzing the self-cleavage activities of various mutants, we determined the base requirements for SSrC as 5'-(U/C/G)-C-N-N-(C/A/G)-(G/A/U)-N-N-3'. PMID- 8223568 TI - Inhibition by the protein kinase C activator 4 beta-phorbol 12-myristate 13 acetate of the prostaglandin F2 alpha-mediated and noradrenaline-mediated but not glucagon-mediated activation of glycogenolysis in rat liver. AB - In perfused rat livers, infusion of prostaglandin F2 alpha (PGF2 alpha) or noradrenaline increased glucose and lactate output and reduced flow. Glucagon increased glucose output and decreased lactate output without influence on flow. Infusion of phorbol 13-myristate 14-acetate (PMA) for 20 min prior to these stimuli strongly inhibited the metabolic and hemodynamic effects of noradrenaline, reduced the metabolic actions of PGF2 alpha but did not alter the effects of glucagon. In isolated rat hepatocytes PGF2 alpha, noradrenaline and glucagon activated glycogen phosphorylase but only PGF2 alpha and noradrenaline increased intracellular inositol 1,4,5-trisphosphate (InsP3). The noradrenaline- or PGF2 alpha-elicited activation of glycogen phosphorylase and increase in InsP3 were largely reduced after preincubation of the cells for 10 min with PMA, whereas the glucagon-mediated enzyme activation was not affected. In contrast to PMA, the phorbol ester 4 alpha-phorbol 13,14-didecanoate, which does not activate protein kinase C, did not attenuate the PGF2 alpha- and noradrenaline-elicited stimulation of glucose output, glycogen phosphorylase and InsP3 formation. Stimulation of InsP3 formation by AlF4-, which activates phospholipase C independently of the receptor, was not attenuated by prior incubation with PMA. Plasma membranes purified from isolated hepatocytes had both a high-capacity, low affinity and a low-capacity, high-affinity binding site for PGF2 alpha. The Kd of the high-capacity, low-affinity binding site was close to the concentration of PGF2 alpha that increased glycogen phosphorylase activity half-maximally. Binding to the high-capacity, low-affinity binding site was enhanced by guanosine 5'-O-(3 thio)triphosphate (GTP[S]). This high-capacity, low-affinity site might thus represent the receptor. The Bmax and Kd of the high-capacity site, as well as the enhancement by GTP[S] of PGF2 alpha binding to this site, remained unaffected by PMA treatment. It is concluded that, in hepatocytes, activation of protein kinase C by PMA interrupted the InsP3-mediated signal pathway from PGF2 alpha via a PGF2 alpha receptor and phospholipase C to glycogen phosphorylase at a point distal of the receptor prior to phospholipase C. PMID- 8223569 TI - Third isoform of the prostaglandin-E-receptor EP3 subtype with different C terminal tail coupling to both stimulation and inhibition of adenylate cyclase. AB - A functional cDNA clone for a third isoform of the mouse prostaglandin-E-receptor EP3 subtype, derived by alternative RNA splicing, named the EP3 gamma receptor, was obtained in addition to those for the two other isoforms, EP3 alpha and EP3 beta. The three isoforms are only different in the amino acid sequence of the putative cytoplasmic carboxy-terminal tail. When expressed, EP3 gamma shows identical ligand-binding properties to these of the other isoforms. The EP3 selective agonist, M&B 28767, increased the basal cAMP level and inhibited the forskolin-induced increase in the cAMP level in EP3 gamma, while it decreased both the basal and forskolin-elevated cAMP levels in EP3 alpha and EP3 beta. The M&B 28767-stimulated GTPase activity consisted of pertussis-toxin-sensitive and cholera-toxin-sensitive portions in the EP3 gamma-expressing cell membrane, suggested that EP3 gamma is coupled to both guanine nucleotide-binding inhibitory and stimulatory proteins. These results indicate that EP3 gamma is coupled to both stimulation and inhibition of adenylate cyclase, but that EP3 alpha and EP3 beta are exclusively coupled to inhibition of adenylate cyclase. Thus, alternative splicing produces a third isoform with a different carboxy-terminal tail, which differs from the other two isoforms in the specificity of coupling to a signal-transduction pathway. PMID- 8223570 TI - Determination of relative binding affinity of influenza virus N9 sialidases with the Fab fragment of monoclonal antibody NC41 using biosensor technology. AB - The relative binding affinities of influenza virus N9 sialidase from term and whale with the Fab fragment of monoclonal antibody NC41 were determined using biosensor technology (Pharmacia BIAcoreTM). The apparent association and dissociation rate constants were measured in real time for the interaction of the Fab with both sialidases, the Fab being immobilised on the sensor surface. Although three-dimensional structural studies have shown that there are no apparent structural differences between the term and whale N9 sialidase epitopes to which the NC41 Fab binds, the apparent binding constant for the interaction with tern N9 sialidase was approximately 2.4-fold higher than that with whale N9 sialidase. The kinetic analysis showed that the association rate constant for the binding of whale N9 sialidase was higher than that for tern N9 sialidase (12.0 x 10(4) M-1 s-1 compared to 4.3 x 10(4) M-1 s-1) and the dissociation rate constants for the whale N9-sialidase-Fab complex were approximately 6-fold higher than for the tern N9-sialidase-Fab complex. Furthermore, kinetic analysis of the dissociation reaction showed that it was composed of two stages, an initial, faster rate followed by a late, slower rate. The values of the relative affinity constants calculated using the initial dissociation rate constant were similar to the values measured at equilibrium in the BIAcore and those determined in true solution equilibrium studies using sedimentation equilibrium. The late, slower, dissociation rate constant yielded affinity constants significantly higher than those obtained by true solution methods. PMID- 8223571 TI - Effect of hydrostatic pressure and cholesterol depletion on the expression of a tumor-associated antigen. AB - The molecular events related to the expression of three tumor-associated epitopes, Ca-MOv17, Ca-MOv18 and Ca-MOv19 have been addressed. The epitopes are carried by a 38-kDa glycoprotein (gp38), recently cloned and identified as a human folate-binding protein. They were found to be coexpressed on the surface of the ovarian carcinoma cell line OVCA432, while they are not coordinately expressed on other adenocarcinoma cell lines (IGROV1, HT-29). This lack of coexpression was investigated from a molecular point of view. We studied three carcinoma cell lines, characterized by a different reactivity with the three relevant monoclonal antibodies MOv17, MOv18 and MOv19. The epitope expression was examined after modifying the membrane properties by using hydrostatic pressure and/or the variation of cholesterol content. Measurement of the expression after cell labelling by mAbs was performed by indirect immunofluorescence, using both fluorescence microscopy and flow cytometry. At variance with HT-29 cells, treatment of ovarian carcinoma IGROV1 cells with hydrostatic pressure failed to exert any effect. On IGROV1, instead, cholesterol depletion affected the expression Ca-MOv17, increasing, in the indirect immunofluorescence tests, the proportion of positive cells from 0 to 66 +/- 9%. Moreover, restoring the cholesterol content of the plasma membrane did not reverse the induced epitope expression. In parallel, immunoprecipitation experiments confirmed that, on IGROV1 surface, gp38 was recognized by all three mAbs. The data presented suggest that in IGROV1 cells the selective lacking of the epitope expression is related to the physical state of the plasma membrane. An explanation is provided by the model of membrane microdomains in which epitope expression may be influenced by the cholesterol level of different plasma membrane regions. PMID- 8223572 TI - Post-transcriptional and post-translational regulatory steps are crucial in controlling the appearance and stability of thylakoid polypeptides during the transition of etiolated tobacco seedlings to white light. AB - We have investigated the expression of nuclear-encoded chloroplast proteins that are not associated with chlorophyll (the lumenal 33-kDa and 23-kDa polypeptides of the oxygen-evolving system of photosystem II, plastocyanin and the Rieske Fe/S protein) by comparing mRNA-accumulation rates with those of the corresponding proteins during illumination of etiolated tobacco seedlings. Using subcellular fractionation, pulse/chase, Northern and Western techniques, we found that the biogenesis and stability of these proteins are regulated both translationally, as well as post-translationally, including the efficiency of mRNA uptake into polysomes, processes that operate between translation and assembly or monitor the status (soluble and membrane-attached) of a terminally processed polypeptide. Polypeptide synthesis is generally not limited by mRNA amounts. For instance, steady-state transcript levels may increase 10-fold during illumination, while those associated with polysomes increase only 2-3-fold without measurable influence on the rate of protein synthesis. The 23-kDa and Rieske polypeptides are predominantly membrane associated, but plastocyanin and the 33-kDa polypeptide are distributed among both soluble and membrane-associated protein fractions. Plastocyanin appears to be comparably stable in both forms. However, for the 33-kDa polypeptide, only the membrane-attached form is stable (> 8 h) and only this pool increases upon illumination. Its soluble form is rapidly degraded with a half-life of approximately 1 h under the chosen conditions. Our findings probably reflect part of a more general regulatory principle operating in the differentiation and maintenance of subcellular structure. PMID- 8223573 TI - Differential regulation of the human H1 zero-histone-gene transcription in human tumor-cell lines. AB - Cloning and sequence analysis of about 2 kb of the 5' flanking region of the human H1 zero histone gene reveals several potential regulatory elements upstream of the transcribed portion of this gene. Transfection studies using the chloramphenicol acetyl transferase (CAT) gene as a reporter gene with a series of promoter deletions revealed that the expression of the H1 zero gene may depend on a complex interplay of several transcription factors, including members of the retinoic acid and/or thyroid-hormone-receptor superfamily, at the 5' flanking region of the H1 zero gene. CAT assays demonstrate varied patterns of expression and regulation in different human tumor-cell lines. The leukemia cell line HL60 does not express H1 zero mRNA and shows no CAT activity. HeLa cells strongly express the CAT gene under the control of the H1 zero promoter. Under the same conditions, HepG2 cells also transcribe the CAT gene, although at a lower rate than HeLa cells. Using different promoter-deletion clones, the CAT activity differs in HepG2 and HeLa cells in the very distal promoter region. In both cell lines, the CAT activity decreases several fold when the region between nucleotides -450 and -600 upstream of the mRNA start site is deleted. It also decreases when just the proximal portion but not the distal promoter region is deleted. In summary, the regulatory patterns of these three cell lines differ, indicating a cell-type-specific regulation of the human H1 zero-histone-gene expression. PMID- 8223574 TI - Immunoelectron microscopic localization of ribosomal proteins BS8, BS9, BS20, BL3 and BL21 on the surface of 30S and 50S subunits from Bacillus stearothermophilus. AB - The locations of ribosomal proteins BS8, BS9 and BS20 on the 30S subunit of Bacillus stearothermophilus ribosomes, and of BL3 and BL21 on the 50S subunit, were determined by immunoelectron microscopy. BL3 was found to lie half-way down the body of the 50S subunit on the interface side, below the L7/L12 stalk, in agreement with the placement of the corresponding protein in Escherichia coli by neutron-scattering; BL21 was located at a similar position on the solvent side of the subunit, as predicted by cross-linking experiments with E. coli ribosomes. Similarly, BS8 was found in the upper region of the body of the 30S subunit on the solvent side, and BS9 on the top of the head of the subunit, also on the solvent side, both positions being in good agreement with neutron-scattering data and other immunoelectron microscopy results. In contrast, BS20 was found to lie at the extreme base of the body of the 30S subunit; this placement is not compatible with the location of E. coli S20 by neutron-scattering but fits very plausibly with other biochemical data, such as sites of RNA-protein footprinting on 16S RNA, relating to the location of S20 in E. coli. PMID- 8223575 TI - Interactions of chloroaluminium-tetramethyl-tetrapyridino-porphyrazine++ + with DNA. AB - The interaction of chloroaluminium-tetramethyl-tetrapyridinoporphyrazine (ClAlTMPyPa) with DNA was investigated by absorption/emission and circular dichroic spectroscopy. Formation of a DNA-porphyrazine complex was evidenced by quenching of the ClAlTMPyPa fluorescence, a hypochromic red shift of the absorption band in the visible part of the spectrum and induction of a characteristic CD band with high positive ellipticity. Scatchard analysis of the spectral changes in low ionic strength buffer suggested an association constant of 9.7 +/- 0.6 x 10(7) M-1. Quenching of the fluorescence of a DNA-ethidium complex by added ClAlTMPyPa was shown to result from direct DNA-dye interaction rather than the displacement of ethidium from DNA. Our data suggest that porphyrazine binds to the outside of the DNA helix, involving interactions which are not limited to electrostatic forces only. The computer assisted modelling of ClAlTMPyPa-DNA interactions showed that the energy-minimized intermolecular orientation involves face-on binding of the ligand, preferably on the minor DNA groove. PMID- 8223576 TI - Glycine decarboxylase complex from higher plants. Molecular cloning, tissue distribution and mass spectrometry analyses of the T protein. AB - cDNA clones encoding the precursor of the T protein of the glycine decarboxylase complex have been isolated from a pea leaf cDNA library in lambda gt11. The longest cDNA insert of 1430 bp encodes a polypeptide of 408 amino acid residues of which 30 residues constitute an N-terminal cleavable presequence and 378 residues make up the mature protein. Several results confirmed the identity of the cDNA and the exactness of the predicted primary structure. Firstly, we purified the T protein to homogeneity and its mass was measured by mass spectrometry. The mass obtained (40966 +/- 5 Da) was the value predicted from the cDNA (40961 Da). Secondly, the purified T protein was chemically cleaved with cyanogen bromide and the peptide fragments were analysed by high-performance liquid chromatography/electrospray ionization mass spectrometry and/or fast-atom bombardment mass spectrometry. The mass values of all the peptides generated by chemical cleavage and measured by these techniques were very close to the values calculated from the predicted primary structure. Thirdly, microsequencing of some of these peptides, which represent 35% of the total protein, fits perfectly with the primary structure deduced from the cDNA. In the present HPLC/electrospray ionization MS studies we never detected the presence of covalently bound tetrahydropteroylpolyglutamate (H4PteGlun), either in the native T protein or in the different peptide fragments generated by the chemical cleavage. The absence of H4PteGlun bound to the T protein in our experimental conditions demonstrates that H4PteGlun is not covalently linked to the T protein. Northern blot analysis showed that the steady-state level of the mRNA corresponding to the T protein was high in green leaves compared to the level in etiolated leaves (approximately 8 10-fold higher). Surprisingly, a non-negligible amount of mRNA corresponding to the T protein was present in roots whereas the mRNA encoding the H protein was not detectable. Western blot analysis showed that the P, L and T proteins of the glycine decarboxylase complex were present in roots whereas the H protein was not detectable. Southern hybridization to pea genomic DNA indicated the presence of a single gene encoding the T protein of the glycine decarboxylase complex in the haploid genome. PMID- 8223577 TI - A mutant T7 phage promoter is specifically transcribed by T7-RNA polymerase in mammalian cells. AB - The phage T7 promoter/polymerase system is highly specific in bacteria in contrast to that observed in mammalian cells. A number of cell lines exhibit a considerable level of expression from the T7 promoter, even in the absence of T7 RNA polymerase. Here, we demonstrate that nuclear-factor-including components of the TFIID fraction, bind to the T7 promoter and inhibit transcription by T7-RNA polymerase. In order to increase the specificity of the promoter for T7-RNA polymerase and to abolish binding of nuclear factors, a novel strategy for the selection of randomly mutated promoters was established. The strategy involves adsorption of mutant promoters to HeLa extracts and binding of the free oligonucleotides to T7-RNA polymerase, cloning, and functional testing of the recombinants. After selection, the resulting mutant promoters showed an increase in specificity for transcription by T7-RNA polymerase. PMID- 8223578 TI - Roulette mutagenesis of the FMN-binding site of Klebsiella pneumoniae flavodoxin. AB - A method of randomising specific regions of coding sequences has been devised which utilises the Lac phenotype to identify mutants. Intact genes can be mutagenised, making it unnecessary to reclone the mutations before examining mutant phenotypes. The method has been applied to three residues around the N terminus of the first alpha helix of the Klebsiella pneumoniae nitrogenase flavodoxin, which are predicted to form part of the phosphate-binding subsite. Surprisingly, most substitutions at Gly12, a highly conserved residue in the chain reversal preceding the alpha helix, appeared to be fairly stable in vivo and were found to retain some function. Substitutions at Lys13, a surface residue which contributes to a patch of positive charge characteristic of the nitrogenase flavodoxins, had no major effect on stability or function. However, most substitutions at Thr14, which is predicted to hydrogen bond to the phosphate of the prosthetic group FMN, were much more destabilising and grossly reduced function. The exceptions were Ala, Cys, Ser and Val, which suggests that the bulk of the residue at this position is critical. PMID- 8223579 TI - The amino acid sequence of human ribonuclease 4, a highly conserved ribonuclease that cleaves specifically on the 3' side of uridine. AB - A ribonuclease (RNase) that cleaves specifically on the 3' side of uridine [Shapiro, R., Fett, J. W., Strydom, D. J. & Vallee, B. L. (1986a) Biochemistry 25, 7255-7264] was purified from human plasma and its amino acid sequence was determined. This protein is a 119-residue single-chain polypeptide cross-linked by four disulfide bonds and has an amino-terminal pyroglutaminyl residue. No post translational modifications were observed during extensive sequence studies on peptide fragments, except for the amino-terminal pyroglutamic acid and a possible deamidation of Asn66. The protein is homologous to the pancreatic ribonucleases and angiogenin, but differs substantially from both of these proteins; the protein sequence has 43% identity with human pancreatic ribonuclease and 39% identity with human angiogenin, as compared to 35% identity between human angiogenin and pancreatic ribonuclease. It is referred to as RNase 4, based on the nomenclature currently used for the genes of pancreatic RNase (RNase 1) and the eosinophil-derived RNases (RNase 2 and RNase 3). Virtually all of the RNase active-site components, including the catalytic residues His12, His119 and Lys41, are preserved. However, some invariant residues of RNase 1 are replaced, e.g. Lys7 by arginine, Asp14 by histidine, and Pro42 by arginine. RNase 4 contains a unique two-residue deletion at the position corresponding to amino acids 77 and 78 of pancreatic RNase, and its carboxyterminal sequence is truncated at position 122. The deletion in angiogenin at position 21 is also found in RNase 4. RNase 4 is very similar to two RNases isolated from bovine and porcine liver, and together they form a new family in the RNase superfamily. The degree of inter species similarity (90%) is much greater than within the pancreatic RNase and angiogenin families, which suggests that this ribonuclease could possess a physiologically important function other than general RNA catabolism. PMID- 8223580 TI - Catalytic and binding properties of restriction endonuclease Cfr9I. AB - The Cfr9I restriction endonuclease recognizes and cleaves duplex DNA sequence C decreases CCGGG. The binding of restriction endonuclease Cfr9I to DNA was examined in the absence of Mg2+ using gel-mobility-shift and nitrocellulose filter-binding assays. It was shown that restriction endonuclease Cfr9I bound DNA fragments either containing or lacking the canonical recognition sequence with equal affinity. These results suggest that the specificity of restriction endonuclease Cfr9I is expressed during the catalytic step. The cleavage of supercoiled pUC18 DNA by restriction endonuclease Cfr9I showed that at low concentrations of MgCl2, only with open-circular DNA, nicks appeared in one strand at the recognition sequence, while the cleavage of the second strand was very slow. At higher concentrations of MgCl2 the enzyme cleaves either one or both strands of the DNA. Under these conditions the supercoiled DNA was converted to open-circular and linear forms simultaneously rather than consecutively. It was shown that open-circular DNA was a poor substrate for restriction endonuclease Cfr9I. These results suggested that both Mg2+ and intact recognition sequence are required to drive the enzyme into correct conformation to ensure DNA cleavage. PMID- 8223581 TI - Identification and characterization of a soluble form of the plasma cell membrane glycoprotein PC-1 (5'-nucleotide phosphodiesterase). AB - PC-1 is a membrane glycoprotein, found on the surface of plasma cells and a few types of nonlymphoid cells, which has recently been found to have 5'-nucleotide phosphodiesterase activity. In this paper, we demonstrate the existence of enzymically active water-soluble forms of PC-1 in ascites from plasmacytoma bearing mice, normal mouse serum, and in supernatants of cultured mouse plasmacytoma cells and mouse L cells transfected with a cDNA encoding the membrane form of PC-1. The water-soluble enzyme activity can be specifically immunoprecipitated by a monoclonal antibody to an allotypic determinant on the membrane form of PC-1, and resides on a slightly smaller polypeptide than membrane PC-1. Biosynthetic studies revealed a single, monomeric, endoglycosidase H-sensitive membrane PC-1 precursor, which was gradually converted to a disulphide-bonded, endoglycosidase-H-resistant form over a period of about 2 h. Soluble PC-1 was first detectable in the supernatant after about 2 h. A distinct intracellular form of soluble PC-1 was not seen. The soluble form of PC-1 does not appear to arise by proteolytic cleavage from the cell surface, although cleavage inside the cell remains a possibility. When taken together with the structure of the relevant portions of PC-1 gene exons, the data suggest that the most likely site of cleavage is between Pro152 and Ala153. PMID- 8223582 TI - Stimulatory effect of NH4+ on the transport of leucine and glucose in an anaerobic alkaliphile. AB - An anaerobic alkaliphile, EP01, specifically requires NH4+ for the acceleration of amino acid and glucose transport [Koyama, N. (1988) FEBS Lett. 253, 187-189]. In this paper, we attempted to clarify how NH4+ is involved in the transport system. The bacterium acidifies the cytoplasm, which was suggested to result in NH4+ accumulation when NH4Cl was added to the medium. Increase of the NH4Cl concentration administered to the medium caused the acceleration of leucine and glucose transport, which was accompanied by an increase in the internal pH and the absolute internal concentration of NH4+, whereas a decrease in the concentration ratio of internal NH4+/external NH4+ was observed. The addition of 3 mM NH4Cl, which resulted in significant stimulation of leucine and glucose transport, raised the internal NH4+ concentration by 42 mM, but the internal pH only by 0.1 units. It seems more likely that leucine and glucose transport are accelerated depending on the increase in the internal NH4+ concentration rather than the increase in the internal pH. By the imposition of an inwardly directed Na+ gradient, the K(+)-loaded membrane vesicles accumulated leucine and glucose, indicating that a sodium chemical potential is available for active transport. The membrane of the bacterium exhibited a Na(+)-stimulated ATPase activity which was remarkably enhanced by the addition of NH4Cl, depending on its concentration, and was inhibited by vanadate. Leucine and glucose transport were inhibited by vanadate. Based on these results, we propose a mechanism in which NH4+ contributes internally to leucine and glucose transport, depending on its concentration, by the activation of a Na(+)-translocating ATPase responsible for the generation of a sodium chemical potential. PMID- 8223583 TI - Role of the rac1 p21-GDP-dissociation inhibitor for rho heterodimer in the activation of the superoxide-forming NADPH oxidase of macrophages. AB - Activation of the superoxide (O2-)-generating NADPH oxidase of phagocytes requires the interaction of membrane-associated cytochrome b559 with three cytosolic components; p47-phox, p67-phox and sigma 1. We proposed that sigma 1 was a heterodimer composed of proteins of 22 kDa and 24 kDa that were tentatively identified as the small GTP-binding protein (G protein) rac1 p21 and GDP dissociation inhibitor for rho (rho GDI). We now describe a modified procedure for the rapid purification of sigma 1 and demonstrate that the NADPH-oxidase activating capacity is associated, throughout the purification sequence, with a protein binding 35S-labelled guanosine 5'-[3-O-thio]triphosphate. SDS/PAGE analysis confirmed the absolute association of sigma 1 activity with the presence of both the 22 kDa and 24 kDa proteins. Immunoblotting with a battery of antibodies against the small G proteins demonstrated that the 22-kDa protein was only recognized by antibodies reacting with rac1 p21; no reaction was found with anti-(rac2 p21), anti-[v-ras(H) p21] and anti anti-(rap1 p21). Free rac1 p21 (not in complex with rho GDI) was not detected at any stage of cytosol fractionation. The proteins comprising the sigma 1 heterodimer could be separated by reverse phase chromatography and amino acid sequencing was performed on peptides derived by trypsin digestion of each of the isolated proteins. This demonstrated the identity of the 22-kDa protein with rac1 p21 and that of the 24-kDa protein with rho GDI. Purified heterodimeric sigma 1 did not require exogenous GTP for activity under conditions that assured the absence of free nucleotides. Treatment of the sigma 1 heterodimer with 1% sodium cholate, followed by gel filtration or anion-exchange chromatography in the presence of 1% sodium cholate, effectively separated rac1 p21 from rho GDI. Monomeric rac1 p21, obtained by these procedures, was able to stimulate cell-free O2- generation. Artificial heterodimeric sigma 1, capable of NADPH oxidase activation, could be reconstituted in vitro by recombining purified monomeric rac1 p21 and rho GDI and removing the sodium cholate used to dissociate the native sigma 1 dimer. Monomeric rac1 p21 exhibited an almost absolute dependence on exogenous GTP following removal of the endogenous nucleotide in low Mg2+ solution. Under similar conditions, heterodimeric sigma 1 was resistant to nucleotide exchange.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8223584 TI - Gene sequence, cDNA construction, expression in Escherichia coli and genetically approached purification of porcine interleukin-1 beta. AB - A genomic clone (PIL3) covering the 8.8-kb prointerleukin-1 beta ('catabolin') gene of the domesticated swine (Sus scrofa domestica) was isolated from a genomic library and characterized by nucleotide sequencing. Typical features of the gene include a seven-exon structure, with the highest degree of nucleotide and amino acid conservation among human and porcine genes being found in the receptor binding portion encoded by exons six and seven. Three 250-bp repetitive elements with a > 75% similarity to the pig repetitive element-1 family sequence are located in untranslated gene segments. Southern-hybridization experiments disclosed extensive genomic heterogeneity of the porcine interleukin-1 beta (IL-1 beta) gene region, suggesting a duplication of at least the 3' half of the gene in the porcine genome. Since similar hybridization patterns were observed for wild boar (Sus scrofa) genomic DNA, it was concluded that this gene rearrangement had preceded domestication of the wild swine. In addition, the cDNA for processed porcine IL-1 beta was constructed through polymerase-chain-reaction-mediated exon fusion by overlap extension starting from the genomic template. Recombinant IL-1 beta was expressed in Escherichia coli as a fusion protein containing an N terminal hexahistidine tag followed by a factor-Xa-cleavage site. The protein was efficiently purified through adoption of a scheme that consisted of four alternating cycles of immobilized metal-ion-affinity chromatography and size exclusion chromatography. 13.8 mg highly purified recombinant porcine IL-1 beta was obtained starting from a 900-ml thermo-induced E. coli culture (final endotoxin concentration < 0.22 ng/ml). The protein behaved homogeneously as a monomeric species, which was reactive in Western-blot experiments with an anti (human-IL-1 beta) serum and which appeared to induce gelatinase B in MDBK cells in a dose-dependent fashion. PMID- 8223585 TI - Biotin biosynthesis in higher plant cells. Identification of intermediates. AB - Biotin biosynthesis was investigated in lavender cell cultures (Lavandula vera L.). Two different biological assays and two different HPLC procedures were used to identify all the intermediates involved in biotin biosynthesis. The pathway for biotin biosynthesis could be analyzed starting with [3H]pimelic acid as precursor, leading to labelled biotin and even to labelled biotinylated enzymes. Intermediates known from the bacterial pathway (7-oxo-8-amino-pelargonic acid, 7,8-diamino-pelargonic acid, dethiobiotin) were present in detectable amounts. Pimelic acid activation to pimeloyl-CoA could be observed. In contrast to bacterial cells, an unknown stable labelled intermediate, named compound A, accumulated. This compound coeluted with an authentic sample of 9 mercaptodethiobiotin from HPLC with an anion-exchange column and was as effective as biotin in supporting the growth of the strain bioB105 of Escherichia coli. When 3H-labelled compound A was added to the growth medium of the lavender cells it was incorporated in an acidomycin-sensitive manner into biotin. [3H]Dethiobiotin was incorporated into both compound A and biotin. These results strongly suggest that, in higher plant cells, the reaction catalysed by biotin synthase may proceed in two distinct steps involving mercaptodethiobiotin (9 mercaptodethiobiotin?) as an intermediate. PMID- 8223586 TI - Specific covalent modification of the tryptophan residues in murine interleukin 6. Effect on biological activity and conformational stability. AB - Modification of recombinant murine interleukin-6 (mIL-6) with the tryptophan specific reagent 2-nitrophenylsulfenyl chloride under mild acidic conditions, 0.1 M sodium acetate, pH 3.5, yielded a derivative containing 2.02 mol 2 nitrophenylsulfenyl tryptophan/mol protein. The sites of modification were identified as Trp36 and Trp160. No detectable side reactions occurred on other amino acids in the molecule, as indicated by the combination of endoproteinase Asp-N peptide mapping, Edman degradation and electrospray mass spectrometry. Sulfenylation of the two tryptophan residues in mIL-6 caused a 50% reduction in both the biological activity in the murine-hybridoma-growth-factor assay using 7TD1 cells and receptor-binding affinity to mIL-6 receptors. Sulfenylation of mIL 6 did not significantly affect the overall conformation of the protein as measured by farultraviolet circular dichroism and binding to the neutralizing anti-mIL-6 mAb 6B4. The sulfenylated protein was, however, significantly less stable [delta delta G(H2O) = 3.98 kJ/mol] than unmodified mIL-6 as measured by urea-gradient gel electrophoresis. PMID- 8223587 TI - Interactions between active-site serine beta-lactamases and so-called beta lactamase-stable antibiotics. Kinetic and molecular modelling studies. AB - The interactions between imipenem and four monobactams and three class A beta lactamases have been studied in detail. Despite their reputation as being beta lactamase-stable, some of these compounds were significantly hydrolysed by the enzymes. The results obtained with the Streptomyces albus G beta-lactamase have been analysed in the light of molecular modelling studies. The discussion is extended to include other so-called beta-lactamase-stable antibiotics to demonstrate that this appellation can often be misleading. PMID- 8223588 TI - A role for protein kinase C-alpha in zymosan-stimulated eicosanoid synthesis in mouse peritoneal macrophages. AB - A possible regulatory function of protein kinase C (PKC) isoenzymes in zymosan stimulated eicosanoid synthesis was studied in mouse peritoneal macrophages in culture. The addition of zymosan to intact cells labelled with [3H]arachidonic acid stimulated a time-dependent and concentration-dependent release of the fatty acid. There was a simultaneous marked increase in the synthesis of prostaglandin E2 and leukotriene C4. The protein-kinase inhibitor K-252a and the selective PKC inhibitor CGP41251 completely blocked zymosan-triggered arachidonic acid release as well as prostaglandin E2 and leukotriene C4 synthesis. In contrast, an inactive staurosporine derivative, CGP42700, failed to inhibit any of the zymosan induced responses. The down-regulation of PKC by long-term treatment with phorbol 12-myristate 13-acetate eliminated zymosan-stimulated arachidonic acid release and eicosanoid synthesis (after 4-6 h treatment). By using specific antibodies it was observed that mouse macrophages express five PKC isoenzymes, PKC-alpha, beta, -delta, -epsilon and -zeta. No PKC-gamma isoenzyme was detected. After exposure to phorbol 12-myristate 13-acetate, a complete depletion of PKC-beta was observed within 1 h and the complete depletion of PKC-alpha and PKC-delta isotypes was observed within 4 h. In contrast, PKC-epsilon was only partially down-regulated after a 24-h treatment with phorbol 12-myristate 13-acetate and PKC-zeta was not affected at all. These data indicate that PKC-alpha and PKC delta isoenzymes are candidates for regulating prostaglandin and leukotriene production. From the potent inhibitory activities of K-252a and CGP41251, two compounds that reportedly display a higher selectivity for PKC-alpha compared to PKC-delta, it is suggested that PKC-alpha triggers arachidonic acid mobilization and eicosanoid synthesis in peritoneal macrophages. PMID- 8223589 TI - Gly85 to Val substitution in pro alpha 1(I) chain causes mild osteogenesis imperfecta and introduces a susceptibility to protease digestion. AB - In this paper we describe a mild moderate form of osteogenesis imperfecta caused by a point mutation in COL1A1 which converted glycine 85 to valine. The valine substitution introduced into the triple-helical domain of type-I collagen a conformational perturbation causing susceptibility to digestive proteases. In fact, SDS/PAGE of pepsin-treated collagen showed the presence of a faint band, migrating between alpha 1(I) and alpha 2(I), both in the medium and in the cell layer. On trypsin digestion the band, a shortened form of alpha 1(I), had a melting temperature of 39.5 degrees C. If the triple-helical collagen was obtained after trypsin or chymotrypsin digestion of procollagen, two shortened bands were identified; the enzymes cleaved about 40% of the trimers. The mutant procollagen was normally secreted and processed in the extracellular matrix at a normal rate. When native type-I collagen was formed after dextran-sulfate incubation, only chains of normal length were found, suggesting that the fibroblast proteases did not recognize the alteration introduced by the mutation. The effects of glycine 85 to valine substitution are compared with those produced by a previously described arginine substitution of the same residue (Deak et al., 1991). PMID- 8223590 TI - Sorbitol dehydrogenase. Full-length cDNA sequencing reveals a mRNA coding for a protein containing an additional 42 amino acids at the N-terminal end. AB - A cDNA clone encoding rat sorbitol dehydrogenase (SDH) was isolated from a rat testis lambda ZAP II cDNA library. The full-length cDNA insert contained 2277 base pairs (bp), starting 182 bp upstream from an ATG codon where translation to the active enzyme SDH is presumed to be initiated. A second ATG codon, however, was found 126 bp upstream, aligned in the same reading frame as that of the active enzyme. Therefore, the coding sequence for SDH can be translated into an additional 42-amino-acid polypeptide linked to the N-terminal amino acid of the enzyme, generating a pre-sorbitol dehydrogenase. The sequence data indicate that the nucleotide environment around this ATG codon is more favorable towards it being the actual open reading frame (ORF) for a pre-SDH than the ATG codon preceding the nucleotide sequence for SDH. Since no known SDH starts with the additional 42 amino acids, it may be that post-translational removal of this polypeptide accompanies the release of the active enzyme. Next, the 3' untranslated region of the cDNA contained a non-coding 1021 bp downstream from the TAA stop codon. The latter sequence included three putative poly(A) signals: one at nucleotides 1362-1367, the second at nucleotides 1465-1470, and the third at nucleotides 2212-2217 [17 bp away from the poly(A) tail]. In addition to the above findings we also report a variance in one of the amino acids in the SDH cDNA sequence. This variance occurs at position 957-960, where threonine is coded for instead of aspartic acid; in the rat testis SDH cDNA, we find the sequence is ACG instead of GAC, as was reported for the rat liver SDH cDNA. Northern-blot hybridization analysis showed that SDH mRNA is a doublet, one band of 4 kb and the other of 2.3-2.4 kb, in both the rat liver and the rat lens, further confirming that the isolated SDH cDNA constituted a full-length cDNA. PMID- 8223591 TI - Correlation between the inhibition of cell growth by accumulated polyamines and the decrease of magnesium and ATP. AB - The mechanism of the antiproliferation effect of spermidine and spermine was studied using a cell culture system of mouse FM3A cells. The addition of either 10 mM spermidine or 2 mM spermine to the growth medium containing 0.9 mM Mg2+ greatly inhibited cell growth (more than 90%). A decrease in the Mg2+ concentration to 50 microM in the growth medium, but without the polyamine addition, did not influence cell growth. However, the concentrations of spermidine and spermine necessary for the inhibition of cell growth when cells were cultured in the presence of 50 microM Mg2+ were much smaller (2 mM spermidine and 0.15 mM spermine). Nevertheless, the amount of polyamines accumulating in cells which could cause the inhibition of cell growth was almost the same, regardless of the large difference in the added polyamine concentrations. At the early stage of polyamine accumulation, the inhibition of cell growth correlated with the decrease of Mg2+ content, but not with a decrease of the ATP content. The decrease in Mg2+ content correlated well with the inhibition of macromolecular synthesis, especially protein synthesis. Thus, the inhibition of cell growth at the early stage of polyamine accumulation was thought to be due to the inactivation of ribosomes through the replacement of Mg2+ on magnesium-binding sites by polyamines. The decrease in Mg2+ content was mainly caused by the inhibition of Mg2+ transport by polyamines. At the later stage of polyamine accumulation, a decrease in ATP content was also observed. This was followed by swelling of the mitochondria, which may be a symptom of the subsequent cell death. PMID- 8223592 TI - In vitro binding of nuclear proteins to the barley plastocyanin gene promoter region. AB - Plastocyanin is a nuclear-encoded chloroplast protein participating in electron transport during photosynthesis. The plastocyanin gene is expressed in photosynthetic tissue in a developmentally regulated manner and the expression is stimulated by light. A genomic clone encoding the plastocyanin precursor was isolated from a barley (Hordeum vulgare) lambda library using a barley cDNA clone as a probe and the sequence of a 1.9-kb DNA fragment containing the plastocyanin gene was determined. TATA and CCAAT boxes are located 34-bp and 68-bp, respectively, upstream of the transcription start site, the 5'-untranslated leader is 78 nucleotides long, and the intronless gene has at least two different polyadenylation sites. DNA sites in the plastocyanin gene that mediate binding of barley nuclear proteins were mapped by mobility-shift assays with fragments of the promoter/upstream region. Two of the three specific binding sites characterised in more detail were found to form complexes with the same factor in cross-competition experiments. One of these sites, narrowed down to a 17-bp sequence at position -512, contains the consensus binding site for Myb-like transcription factors. The third specific binding site, located at position -622, contains the sequence CACGTG which is a high-affinity-binding site for transcription factors of the basic-region leucine-zipper family. PMID- 8223593 TI - Proof-reading 3'-->5' exonucleases isolated from rat liver nuclei. AB - Mammalian nuclear DNA polymerases alpha and beta are known to be devoid of the editing 3'-->5' exonucleolytic activity. Presumably this activity could be effected by the exonucleases non-associated covalently with DNA polymerases. Two 3'-->5' exonucleases of 40 kDa and 50 kDa (exo-40 and exo-5) have been isolated from rat liver nuclei and purified to near homogeneity. They are shown to excise mismatched nucleotides from poly[d(A-T)] template, respectively, 10-fold and 2 fold faster than the matched ones. Upon addition of either of these exonucleases to the DNA polymerase alpha from rat liver or calf thymus, the fidelity of in vitro reproduction of the primed DNA from bacteriophage phi X174 amber 3 is increased 5-10-fold, levels of exonuclease and DNA-polymerase activities being similar. Extrapolation of in-vitro DNA-replication fidelity to the cellular levels of activities of the exonucleases and the alpha-polymerase suggests that exonucleolytic proof-reading augments the accuracy of DNA synthesis by 2-3 orders of magnitude. PMID- 8223594 TI - Identification of a 100-kDa protein associated with nuclear ribonuclease P activity in Schizosaccharomyces pombe. AB - Ribonuclease P from the fission yeast Schizosaccharomyces pombe has been purified to apparent homogeneity. A purification of 23,000-fold was achieved by four fractionation steps with DEAE-cellulose chromatography, phosphocellulose chromatography, glycerol-gradient fractionation and finally tRNA-affinity chromatography. A 100-kDa protein was present in the most pure preparations in amounts approximately stoichiometric with the previously identified RNA components of the enzyme, K1-RNA and K2-RNA [Krupp, G., Cherayil, B., Frendeway, D., Nishikawa, S. & Soll, D. (1986) EMBO J. 5, 1697-1703]. A cross-linking experiment utilizing a 4-thiouridine-substituted precursor tRNA demonstrated that the 100-kDa protein interacts with the ribonuclease P substrate in a specific fashion. We therefore conclude that the protein component of S. pombe ribonuclease P is a 100-kDa protein. PMID- 8223595 TI - Specific molecular interaction sites on factor VII involved in factor X activation. AB - Factor VII, a serine-protease zymogen, and tissue factor, the cellular receptor/coenzyme, are the protein components of the macromolecular complex which initiates the extrinsic pathway of the coagulation cascade. Previous studies were directed to the identification of functional sites on factor VII which mediate factor X activation, employing a series of potentially inhibitory synthetic peptides representing the primary structure of factor VII and antibodies to selected peptides. The involvement of at least four high-affinity interactive regions [factor VII (44-50), (196-229), (285-305) and (376-396) peptides] on the surface of factor VII was clearly demonstrated. The minimal sequences for the expression of inhibitory activity of these four molecular recognition domains on factor VII were identified using short and overlapping peptides. The short factor VII-(206-218)-peptide (most inhibitory peptide in the sequence 196-229 on factor VII) inhibited the binding of factor VII to the tissue-factor-expressing J82 cell line. Furthermore, radiolabeled [Tyr201] factor VII-(199-221)-peptide, with a tyrosine substituted for the normal tryptophan residue, was specifically bound to J82 cells, and also the binding of the radiolabeled peptide to this cell line was specifically inhibited by a monoclonal antibody to tissue factor, confirming that the interaction site for tissue factor on factor VII is present within the peptide sequence 196-229. Kinetic analyses suggested that the regions represented by factor VII-(285-305)- and factor VII-(376-396)-peptides are involved in factor X recognition and the chemical cross-linking of the radiolabeled peptides resulted in specific binding to factor X, confirming that these two regions on factor VII represent the substrate-recognition site. Furthermore, these radiolabeled peptides specifically interact with the heavy chain of factor X, suggesting that the complementary binding region for the substrate-recognition site on factor VII are present on the heavy chain of factor X. PMID- 8223596 TI - Limited proteolysis of yeast phosphofructokinase. Sequence locations of cleavage sites created by the actions of different proteinases. AB - Purified phosphofructokinase 1 from baker's yeast (Saccharomyces cerevisiae) was subjected to proteolysis by thermolysin, endoproteinase lys-C, trypsin and chymotrypsin under defined solvent conditions. In the absence of substrates and allosteric effectors, the catalytic activity of phosphofructokinase rapidly disappeared in the presence of each proteolytic enzyme. The presence of a saturating concentration of ATP protected phosphofructokinase activity from proteolytic inactivation while the collective presence of fructose 6-phosphate, AMP and fructose 2,6-bisphosphate provided transient activation during proteolysis. Changes in the quaternary structure of phosphofructokinase resulting from proteolysis were estimated by high performance size exclusion chromatography while changes in the primary sequence of the individual alpha and beta polypeptide chains were estimated by polyacrylamide-gel electrophoresis in sodium dodecylsulfate. The site(s) of proteolytic cleavage were identified by N-terminal sequence analysis of resolved electrophoretic components. The presence of ATP protects phosphofructokinase from thermolysin proteolysis, while the collective presence of fructose 6-phosphate, AMP and fructose 2,6-bisphosphate restricts proteolysis to one site in each polypeptide chain involving the peptide bonds preceding Leu199 in the alpha chain and Leu192 in the beta chain. The truncated phosphofructokinase retains its octameric structure. The presence of ATP largely restricts endoproteinase lys-C proteolysis to a single site in the alpha chain involving the peptide bond preceding Val914. This cleavage results in the dissociation of the octameric form of phosphofructokinase into two tetramers. The presence of ATP restricts both trypsin and chymotrypsin proteolysis to the N terminal and C-terminal regions described above, resulting in the preferential stabilization of the tetrameric form of phosphofructokinase. It would appear that the first 200 and last 80 residues which are unique to the sequence of the yeast phosphofructokinase are not directly involved in catalysis or its allosteric regulation. However, the last 80 residues of the alpha polypeptide chain do appear to stabilize an octameric structure which is unique to yeast phosphofructokinase. PMID- 8223597 TI - Molecular cloning and primary structure of Man9-mannosidase from human kidney. AB - Man9-mannosidase, a processing enzyme found in the endoplasmic reticulum (ER), catalyses the removal of three distinct mannose residues from peptide-bound Man9 GlcNAc2 oligosaccharides producing a single Man6 isomer [Bause, E., Breuer, W., Schweden, J., Roesser, R. & Geyer, R. (1992) Eur. J. Biochem. 208, 451-457]. We have isolated four Man9-mannosidase-specific clones from a human kidney cDNA library and used these to construct a full-length cDNA of 3250 base pairs. A single open reading frame of 1875 nucleotides encodes a protein of approximately 71 kDa, consistent with data from immunological studies. Analysis of the coding sequence predicts that Man9-mannosidase is a type II transmembrane protein consisting of a short cytoplasmic polypeptide tail, a single transmembrane domain acting as a non-cleavable signal sequence and a large luminal catalytic domain. This domain architecture closely resembles that of other ER and Golgi-located processing enzymes, pointing to common structural motifs involved in membrane insertion and topology. The protein sequence of the Man9-mannosidase contains three potential N-glycosylation sites of which only one site is used. The amino acid sequence of several peptide regions, including a calcium-binding consensus sequence, bears striking similarities to an ER alpha-1,2-mannosidase from yeast, whereas, by contrast, no sequence similarity was detectable with rat liver ER alpha-mannosidase and Golgi alpha-mannosidase II. This finding may indicate that the mammalian alpha-mannosidases, which differ significantly in their substrate specificity, are coded for by evolutionarily unrelated genes, providing an attractive means of regulation and fine-tuning oligosaccharide processing, not only at the enzymic but also at the transcriptional level. PMID- 8223598 TI - Tyrosinase isoenzymes in mammalian melanocytes. 1. Biochemical characterization of two melanosomal tyrosinases from B16 mouse melanoma. AB - B-16 mouse melanoma melanosomes contain two forms of tyrosinase that can be resolved by SDS/PAGE. These forms interact to different extents with the ion exchanger DEAE-Sephadex and with hydroxyapatite, and have different affinity for the melanosomal membrane and/or the intraorganular matrix. After partial purification and complete separation of the two tyrosinases, several kinetic parameters were analyzed. The form of lower electrophoretic mobility displayed a higher Km for 3,4-dihydroxy-L-phenylalanine (L-dopa) and L-tyrosine, an absolute requirement for the cofactor L-dopa in its tyrosine hydroxylase activity, and a lower ratio of tyrosine hydroxylation to Dopa oxidation. The form of higher electrophoretic mobility displayed lower values of Km for both substrates and was able to exhibit tyrosine hydroxylase activity after a lag period even in the absence of L-dopa. Both forms were stereospecific for the L isomers and sensitive to the specific tyrosinase inhibitor 2-phenylthiourea. These forms do not appear to result from different degrees of glycosylation, nor from limited proteolysis and are also present in the microsomal fraction of B16 mouse melanoma. They might correspond to different gene products, most likely derived from the b and c loci. PMID- 8223599 TI - Purification and characterization of endoglucanase C from Clostridium cellulolyticum. Catalytic comparison with endoglucanase A. AB - An Escherichia coli clone was constructed to overproduce endoglucanase C (CelCCC) from Clostridium cellulolyticum. This construction made it easier to isolate the enzyme but, as observed in the case of endoglucanase A (CelCCA) from the same organism, the purification led to the isolation of two forms of the cellulase differing in their molecular masses, 48 kDa and 41 kDa. N-terminal sequence analysis of both purified enzymes showed that the shorter form was probably the result of partial proteolysis near the COOH-extremity. The difference in mass indicated that the shorter protein lacks the C-terminal reiterated domains (20-24 amino-acid twice-repeated sequences). These particular domains are characteristic of clostridial cellulases acting on cellulose by the mean of cellulosomal particles. Biochemical and enzymic studies were performed on each form of CelCCC, and revealed that their temperature and pH optima were identical, but their catalytic parameters were quite different. Furthermore, the differences of enzymic behavior observed between the two forms of CelCCC are almost identical to those already noted in the case of the two forms of CelCCA. The stereoselectivity of the reaction catalysed by CelCCC and CelCCA was determined using proton NMR spectroscopy; CelCCC acts by configuration inversion, whereas CelCCA acts by configuration retention. The degradation patterns on cellodextrins (ranging from cellotriose to cellohexaose) and chromophoric cellodextrins (from p-nitrophenyl cellobiose to p-nitrophenyl-cellopentaose) were also investigated in both forms of CelCCC and CelCCA. It emerged that the natural cellodextrins degradation patterns of CelCCC and CelCCA were very similar but the utilization of p nitrophenyl-cellodextrins showed the existence of considerable differences between these two endoglucanases in terms of cleavage-site position and catalytic parameters. CelCCC and CelCCA were found not to act synergistically on the tested substrates. PMID- 8223600 TI - Purification, nucleotide sequence and some properties of a bifunctional isomerase/decarboxylase from the homoprotocatechuate degradative pathway of Escherichia coli C. AB - A 1.8-kbp region of DNA that appeared from deletion subcloning to code for 2 hydroxyhepta-2,4-diene-1,7-dioate isomerase and 5-oxopent-3-ene-1,2,5 tricarboxylate decarboxylase was investigated further. By nucleotide sequencing, a single open reading frame was found encoding a polypeptide of M(r)44514. One of the deletion subclones expressed the decarboxylase and isomerase activities at elevated levels and was used to facilitate purification of the enzyme(s). Both activities copurified, indicating that they were distinct activities of the same protein. Some kinetic properties of the purified isomerase/decarboxylase protein were investigated and it was shown that there is a 49,000-fold preference for 2 hydroxyhepta-2,4-diene-1,7-dioate over the structurally related compound 5 carboxymethyl-2-hydroxymuconate, the substrate of a second isomerase in the same catabolic pathway. Comparison of the amino acid sequences of the two isomerases showed only a low level of similarity, suggesting that these two enzymes are not evolutionarily related. However, comparison of the N-terminal half of the isomerase/decarboxylase sequence (residues 1-202) with the second half (residues 203-406) showed significant similarity, suggesting that a duplication may have occurred to produce the bifunctional gene. PMID- 8223601 TI - Structural flexibility of the calmodulin-binding locus in Bordetella pertussis adenylate cyclase. Reconstitution of catalytically active species from fragments or inactive forms of the enzyme. AB - The catalytic domain of Bordetella pertussis adenylate cyclase, a calmodulin activated enzyme with toxic properties, is a modular construct cleaved by trypsin into two subdomains of 224 (T25) and 175 (T18) amino acids. The calmodulin binding locus of the bacterial enzyme consists of approximately 70 amino acids and overlaps the C-terminus of T25 and the N-terminus of T18. This region, exposed to the solvent or proteases, also exhibits an unusual high flexibility and allows, as demonstrated in this study, reconstitution in the presence of calmodulin of active species of adenylate cyclase from overlapping inactive fragments of the enzyme. Moreover, several combinations of inactive variants of the bacterial enzyme obtained by site-directed mutagenesis can yield active species. Heterodimers, resulting from a few selected combinations of inactive species of adenylate cyclase, exhibit specific activity similar to that of the native enzyme. Productive complementation from inactive fragments is a unique phenomenon among calmodulin-activated enzymes and represents a new and helpful tool in the understanding of the molecular mechanism of activation of B. pertussis adenylate cyclase upon binding of calmodulin. PMID- 8223602 TI - Properties of the two isoenzymes of methyl-coenzyme M reductase in Methanobacterium thermoautotrophicum. AB - Methyl-coenzyme M reductase (MCR) catalyses the methane-forming step in the energy metabolism of methanogenic Archaea. It brings about the reduction of methyl-coenzyme M (CH3-S-CoM) by 7-mercaptoheptanoylthreonine phosphate (H-S HTP). Methanobacterium thermoautotrophicum contains two isoenzymes of MCR, designated MCR I and MCR II, which are expressed differentially under different conditions of growth. These two isoenzymes have been separated, purified and their catalytic and spectroscopic properties determined. Initial-velocity measurements of the two-substrate reaction showed that the kinetic mechanism for both isoenzymes involved ternary-complex formation. Double reciprocal plots of initial rates versus the concentration of either one of the two substrates at different constant concentrations of the other substrate were linear and intersected on the abcissa to the left of the 1/v axis. The two purified isoenzymes differed in their Km values for H-S-HTP and for CH3-S-CoM and in Vmax. MCR I displayed a Km for H-S-HTP of 0.1-0.3 mM, a Km for CH3-S-CoM of 0.6-0.8 mM and a Vmax of about 6 mumol.min-1 x mg-1 (most active preparation). MCR II showed a Km for H-S-HTP of 0.4-0.6 mM, a Km for CH3-S-CoM of 1.3-1.5 mM and a Vmax of about 21 mumol.min-1 x mg-1 (most active preparation). The pH optimum of MCR I was 7.0-7.5 and that of MCR II 7.5-8.0. Both isoenzymes exhibited very similar temperature activity optima and EPR properties. The location of MCR I and of MCR II within the cell, determined via immunogold labeling, was found to be essentially identical. The possible basis for the existence of MCR isoenzymes in M. thermoautotrophicum is discussed. PMID- 8223603 TI - The glycosylphosphatidylinositol anchor of the trypomastigote-specific Tc-85 glycoprotein from Trypanosoma cruzi. Metabolic-labeling and structural studies. AB - The Tc-85 glycoprotein, specific for the infective stage of Trypanosoma cruzi, is anchored via glycosylphosphatidylinositol. The protein was purified from parasites, labeled metabolically with palmitic acid, by immunoprecipitation with the H1A10 monoclonal antibody or by affinity column chromatography on wheat germ agglutinin. Antisera to the soluble form of the variant surface glycoprotein of Trypanosoma brucei brucei cross-reacted with Tc-85 when the immunoprecipitate was analysed by Western blotting. The reaction was intensified upon previous incubation of the glycoprotein with phosphatidylinositol-specific phospholipase C. Such recognition was abolished when the cyclic phosphate was opened by mild acid treatment. The lipid cleaved by phospholipase C digestion, was identified as 1-O-hexadecylglycerol by reverse-phase thin-layer chromatography. The glycan core was deaminated and chemically labeled by reduction with NaB3H4. The labeled glycoprotein was exhaustively treated with pronase and dephosphorylated with 50% HF. Although microheterogeneity of the oligosaccharide moiety was apparent, by thin layer chromatography, a main spot coincident with Man(alpha 1-2) Man(alpha 1 6) Man(alpha 1-4) anhydromannitol was shown, consistent with the conserved core structure of all glycosylphosphatidylinositol anchors analysed to date. PMID- 8223604 TI - Different organization of the tRNA-gene-associated repetitive element, DRE, in NC4-derived strains and in other wild-type Dictyostelium discoideum strains. AB - The retrotransposon DRE (Dictyostelium repetitive element) was discovered in the course of an extensive study concerning the genomic organization of tRNA genes in the NC4-derived strains AX2 and AX3 of the cellular slime mold Dictyostelium discoideum. As a striking feature, DRE was found exclusively in a constant orientation and at a constant distance upstream from different tRNA genes. About 150-200 DRE with intact 5'-terminal-repeat structures are present in NC4-derived strains. These strains were termed high-copy DRE strains (HCD strains) as opposed to low-copy DRE strains (LCD strains) such as the wild-type D. discoideum isolates DD61, WS380B, OHIO and V12. LCD strains contain only 3-15 DRE with intact 5'-terminal-repeat-structures. However, in addition to these few intact elements, many 5'-truncated DRE elements are present in LCD strains. In HCD strains, most DRE show typical structural characteristics of retrotransposons containing terminal repeats at both ends, which seems to be one prerequisite for active transposition. In LCD strains, however, most DRE elements are 5' truncated, which is a common feature of eukaryotic LINE elements. Despite their truncated 5'-ends, DRE in LCD strains retain unique integration specificities, i.e. they are always found position-specifically and orientation-specifically integrated in front of tRNA genes, flanked by a 12-16-bp target-site duplication. PMID- 8223605 TI - Transcriptional control of the heme oxygenase gene during mouse spermatogenesis. AB - The activation of the heme oxygenase (HO) gene during development of mouse male germ cells was examined by nuclear run-off assay. Results showed that the HO gene was activated in male germ cells, especially in spermatogonia and that the mRNA was stored throughout meiosis. In contrast, Western blotting analysis revealed that HO was expressed during spermatogenesis to spermatocytes. As the mouse HO gene is known to have a 12-o-tetradecanoyl-phorbol 13-acetate(TPA)-responsive element (TRE) in its upstream region and is activated by TPA in mouse M1 cells, we also examined the activation of the nuclear proto-oncogenes fos and jun, which are activators of TRE. Both genes were found to be activated with expression of their protein products only in spermatogonia. Moreover activation of fos, jun and HO genes in spermatogonia was strongly inhibited by a c-kinase inhibitor. These results suggest that the HO gene is activated at the spermatogonia stage by a fos/jun heterodimer complex (AP1) through c-kinase activation. PMID- 8223606 TI - Solution conformation of an antibacterial peptide, sarcotoxin IA, as determined by 1H-NMR. AB - The solution conformation of sarcotoxin IA, which is an antibacterial peptide isolated from Sarcophaga peregrina with a molecular mass of 4 kDa, was determined by NMR spectroscopy and hybrid distance geometry/dynamical simulated annealing calculations. On the basis of 227 experimental constraints, including 185 distance constraints obtained from NOE and 42 constraints associated with 21 hydrogen bonds, a total of 18 converged structures of sarcotoxin IA were obtained. The final 18 converged structures exhibit backbone-atomic root-mean square differences about the averaged coordinate positions of 0.070 +/- 0.027 nm for residues 3-23 and 0.040 +/- 0.017 nm for residues 28-38. It has been indicated that sarcotoxin IA consists of two amphiphilic alpha-helical regions, i.e. helix I (Leu3-Gln23) and helix II (Ala28-Ala38), with a hinge region (Gly24 Ile27), which connects helix I and helix II. We conclude that these two amphiphilic helical segments of sarcotoxin IA are of importance for the expression of the antibacterial activity. PMID- 8223607 TI - Interactions of calcium and magnesium with the mitochondrial inorganic pyrophosphatase from Saccharomyces cerevisiae. AB - The activity of the mitochondrial inorganic pyrophosphatase from Saccharomyces cerevisiae was measured in the presence of increasing concentrations of magnesium and calcium. Calcium pyrophosphate (dissociation constant Kd = 1.9 microM) inhibited pyrophosphatase by competition with magnesium pyrophosphate (Kd = 50 microM). The small movements of calcium detected in mitochondria from yeast may be physiologically significant for the control of inorganic pyrophosphatase activity and the concentration of pyrophosphate in the matrix of yeast mitochondria. PMID- 8223608 TI - Aggregation of pyrene-labeled microsomal glutathione S-transferase. Effect of concentration. AB - Microsomal glutathione S-transferase was labeled by the fluorescence probe N-(1 pyrenyl)maleimide which modified 1 mol thiol residue/mol protein. The enzyme activity increased about tenfold after the binding. The pyrene-labeled microsomal glutathione S-transferase exhibited two fluorescence bands which are typical of pyrene; one at 393 nm attributable to unassociated pyrenes, the other at 480 nm attributable to pyrene excimers (excited dimers). The excimeric fluorescence increased at high protein concentrations indicating a shift of the equilibrium of labeled polypeptide chains from trimeric complexes, the functional unit of microsomal glutathione S-transferase, to larger aggregates. At 25 degrees C and at a 1% Triton X-100 concentration, the calculated equilibrium constant of this process is 65 microM. Along with the formation of large aggregates, a progressive increase of the enzymic activity was observed. Thus, N-(1-pyrenyl)maleimide appears to be a very useful probe to study the supramolecular structure of this enzyme. PMID- 8223609 TI - Accessibility to modification of histidine residues of RecA protein upon DNA and cofactor binding. AB - The potential role of histidine residues of RecA protein in binding DNA has been investigated by monitoring their accessibility to diethylpyrocarbonate. In the absence of both DNA and cofactor, only one of two histidine residues is modified by the reagent, indicating that the other residue is buried. However, both histidine residues become accessible after addition of cofactor analog adenosine 5'-O-(3-thiotriphosphate) (ATP[S]) indicating a change in the organization of the RecA filament and/or a change in the conformation of protein. The diethylpyrocarbonate-modified RecA is found to be able to polymerize just as the unmodified protein. The binding of double-stranded DNA, in the presence of ATP[S], reduces the reactivity of both histidine residues to diethylpyrocarbonate. The binding of single-stranded DNA (with ATP[S]) has a similar, though smaller, protective effect. However, no significant dissociation of either of the complexes as a result of the modification was observed and a RecA molecule which had been modified in the absence of DNA could still bind DNA. A protection of the histidine residues is also effected by high salt concentration which promotes, just as DNA binding, ATPase and coprotease activity in RecA. The protection of histidine residues to diethylpyrocarbonate upon DNA binding probably relates to a conformational change of RecA and may not be any direct effect of shielding by the DNA. Nonetheless, the domains including the histidine residues could be centers of allosteric effects and are concluded to be close to the DNA binding site. PMID- 8223610 TI - Increased levels of alpha-class and pi-class glutathione S-transferases in cell lines resistant to 1-chloro-2,4-dinitrobenzene. AB - Glutathione S-transferase (GST) enzymes are often over-expressed in tumor cells made resistant to cytotoxic drugs but it is unclear whether GST over-expression is directly linked to the resistance mechanism. We have made a human lung tumor cell line resistant to 1-chloro-2,4-dinitrobenzene (CDNB) in order to establish whether selection for resistance with a model GST substrate results in selection of a cell line with higher GST levels. The resistant line (CDNBr), although only twofold more resistant to this compound, exhibited a marked (15-fold) increase in GST activity compared to the wild-type cell line (28 +/- 10 versus 357 +/- 30 nmol CDNB conjugated.min-1 x mg-1 protein, respectively). Resistance to CDNB was associated with a marked increase in the level of both alpha-class and pi-class GST. Resolution of the GST by reverse-phase HPLC demonstrated that the increase in the expression of the alpha-class enzymes was due to elevated levels of both the B1 and B2 subunits. The increased levels of alpha-class and pi-class GST in the CDNBr cells was not due to either gene amplification or increased mRNA levels and appears to involve either altered mRNA utilization or protein stabilization. In addition to being resistant to CDNB, the CDNBr cell line also showed a 2.5 fold resistance to cumene hydroperoxide but was not cross-resistant to the anticancer drug chlorambucil. To demonstrate that the increased GST level was part of the resistance mechanism the alpha-class GST B1 cDNA under control of the beta-actin promoter was stably expressed in the breast tumor cell line MCF-7. The cell lines generated were twofold more resistant to CDNB relative to the parental line. PMID- 8223611 TI - The alpha-mannosyl-binding lectin from leaves of the orchid twayblade (Listera ovata). Application to separation of alpha-D-mannans from alpha-D-glucans. AB - The carbohydrate-binding specificity of an alpha-D-mannose-specific lectin isolated from leaves of the orchid twayblade (Listera ovata) was elucidated by quantitative precipitation of mannose-containing polysaccharides and glycoproteins, hapten inhibition, and affinity chromatography on the immobilized lectin. L. ovata agglutinin (LOA) interacted with various alpha-mannans and galactomannans of yeasts, fungi and bacteria, but not with alpha-glucans, e.g., dextran and glycogen, as do mannose/glucose-binding lectins. This lectin, LOA, appears to be highly specific for alpha 1-3 mannosidic linkages. It reacted with a linear alpha 1-3-mannan (D. P. 15) and, surprisingly, even with a linear alpha 1-3-mannoheptasaccharide. The LOA/C. tropicalis mannan precipitation reaction was inhibited by alpha-linked mannooligosaccharides, in the order, alpha 1-3 > alpha 1-6 > alpha 1-2 linkages; alpha 1-3 [Man]4 and [Man]5 were the best inhibitors among various mannooligosaccharides tested, having 7-times greater potency than alpha 1-3 [Man]2, and 18-times that of methyl, alpha-mannoside. LOA/mannan interaction was also inhibited by periodate-oxidized and reduced alpha 1-3 [Man]5 which had an inhibitory potency similar to that of alpha 1-3 [Man]3, confirming that LOA also recognizes the internal alpha 1-3-mannosidic linkages of carbohydrate chains. Complete resolution of mannan and glycogen from yeast cells, by affinity chromatography on an immobilized LOA column, and retention of several high-mannose-glycoproteins suggest this lectin to be a useful tool for purification and structural investigation of alpha-mannosyl-containing polysaccharides and glycoconjugates. PMID- 8223612 TI - Repression of histone gene transcription in quiescent 3T6 fibroblasts. AB - Maintaining murine 3T6 fibroblasts in serum-depleted medium for a period of three days results in a resting cell population that does not synthesize DNA. Histone mRNA levels, closely tied to the cell-proliferation rate, are low due to a reduced rate of synthesis. A comparison of histone gene transcription in vitro by nuclear extracts of quiescent or proliferative 3T6 cells showed that a 200-bp segment of the promoter was responsible for repressing gene activity when cells were in a G0 state. In the absence of the distal promoter region (-200 to -400), gene transcription remained high in quiescent cells, indicating the proximal promoter region (+1 to -200) was responsible for basal gene activity. Alterations in protein binding to the distal promoter region correlated with histone H4 gene activity, suggesting that repression of histone gene transcription is linked to the attachment of a specific nuclear protein. During G1, the histone H4 gene was efficiently transcribed in vitro, but an inability to process the histone pre mRNA limited the cellular content of mature histone mRNA. This distinction between transcriptional (in G0) and post-transcriptional (in G1) mechanisms for modulating histone mRNA levels suggests that gene-regulatory factors are specifically activated in quiescent cells to reduce expression of non-essential genes. PMID- 8223613 TI - Disruption of the Kluyveromyces lactis GGS1 gene causes inability to grow on glucose and fructose and is suppressed by mutations that reduce sugar uptake. AB - In the yeast Saccharomyces cerevisiae the GGS1 gene is essential for growth on glucose or other readily fermentable sugars. GGS1 is the same gene as TPS1 which was identified as encoding a subunit of the trehalose-6-phosphate synthase/phosphatase complex and it is allelic to the fdp1, byp1, glc6 and cif1 mutations. Its precise function in the regulation of sugar catabolism is unknown. We have cloned the GGS1 homologue from the distantly related yeast Kluyveromyces lactis. The KlGGS1 gene is 74% and 79% identical at the nucleotide and amino acid sequence level, respectively, to the S. cerevisiae counterpart. We also compared the sequence with the partly homologous products of the S. cerevisiae genes TPS2 and TSL1 which code for the larger subunits of the trehalose synthase complex and with a TSL1 homologue, TPS3, of unknown function. Multiple alignment of these sequences revealed several particularly well conserved elements. Disruption of GGS1 in K. lactis caused the same pleiotropic phenotype as in S. cerevisiae, i.e. inability to grow on glucose or fructose and strongly reduced trehalose content. We have also studied short-term glucose-induced regulatory effects related to cAMP and cAMP-dependent protein kinase, i.e. the cAMP signal, trehalase activation, trehalose mobilization and inactivation of fructose-1,6 bisphosphatase. These effects occur very rapidly in S. cerevisiae and are absent in the Scggs1 mutant. In K. lactis all these effects were much slower and largely unaffected by the Klggs1 mutation. On the other hand, glucose strongly induced pyruvate decarboxylase and activated the potassium transport system in K. lactis and both effects were absent in the Klggs1 mutant. Addition of glucose to galactose-grown cells of the Klggs1 mutant caused, as in S. cerevisiae, intracellular accumulation of free glucose and of sugar phosphates and a rapid drop of the ATP and inorganic phosphate levels. Glucose transport kinetics were the same for the wild type and the Klggs1 mutant in both derepressed cells and in cells incubated with glucose. We have isolated phenotypic revertants of the Klggs1 mutant for growth on fructose. The suppressors that we characterized had, to different extents, diminished glucose uptake in derepressed cells but cells incubated in glucose showed very different characteristics. The suppressor mutations prevented deregulation of glycolysis in the Klggs1 mutant but not the accumulation of free glucose. The mutants with higher residual uptake activity showed partially restored induction of pyruvate decarboxylase and activation of potassium transport.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8223614 TI - The phospholipase-A2 reaction leads to increased monocyte adhesion of endothelial cells via the expression of adhesion molecules. AB - Mononuclear cell invasion into the vascular-vessel wall is a very important initial step in the development of atherosclerotic lesions. Hypercholesterolemia leads to a marked adhesion of circulating blood monocytes to arterial endothelial cells in vivo, and minimally oxidized low-density lipoprotein enhances monocyte adhesion to endothelial cells in vitro. The activation of phospholipase A2 (PLA2) is also important in the oxidation of low-density lipoprotein by endothelial cells. In this study, we investigated the role of PLA2 activation in the adhesion of a leukemic monocyte cell line (THP-1 cells) to endothelial cells in vitro using an adhesion assay and a cell-ELISA technique. The treatment of human umbilical-cord-vein endothelial cells with PLA2 stimulators such as interleukin-1 beta, tumor necrosis factor and lipopolysaccharide all increased the adhesion of THP-1 cells to endothelial cells. Exogenous PLA2 also increased the adhesion of these cell types. The increased adhesion induced by these PLA2 stimulators, as well as PLA2 itself, was reversed by various inhibitors of the PLA2 reaction. A product of the PLA2 reaction, lysophosphatidylcholine, also increased cell adhesion. A cell-ELISA technique showed the enhanced expression of vascular-cell adhesion-molecule 1 and intercellular-adhesion-molecule 1 to endothelial cells after treatment with PLA2 stimulators, PLA2 or lysophosphatidylcholine. These results suggest that the PLA2 reaction enhances monocyte adhesion to endothelial cells through the expression of cellular adhesion molecules. PMID- 8223615 TI - Putative dehydrogenase tms1 suppresses growth arrest induced by a p53 tumour mutant in fission yeast. AB - Expression of a human tumour-derived p53 His 273 cDNA induced growth arrest in fission yeast Schizosaccharomyces pombe. Based on the p53-induced growth arrest, we cloned an extragenic suppressor, termed tms1, by complementation. The open reading frame of the tms1 gene corresponded to a protein of 347 amino acids with a calculated mass of 37380 Da. The transcriptional start site of the tms1 gene was mapped and, in addition, the corresponding cDNA was isolated and expressed in Escherichia coli. Recombinant tms1 protein served as an antigen to produce specific polyclonal antibodies to aid identification of the tms1-gene-product in total yeast lysates. Comparison of the deduced amino acid sequence of tms1 with available databases revealed significant similarity to dehydrogenases, suggesting that the tms1 protein itself might possess dehydrogenase activity. PMID- 8223616 TI - Intramolecular distances within the Ca(2+)-ATPase from sarcoplasmic reticulum as estimated through fluorescence energy transfer between probes. AB - Fluorescence energy transfer measurements have been carried out to estimate intramolecular distances between probes bound to Ca(2+)-transporting ATPase (Ca(2+)-ATPase) as well as distances between these probes and the phospholipid headgroup. The nucleotide binding site was monitored by using 1,N6 ethenoadenosine 5'-triphosphate, a fluorescent analogue of ATP, and also by labelling Lys515 with fluorescein 5'-isothiocyanate. Three different cysteine residues were individually labelled using the following probes: 5-[(2 iodoacetyl)aminoethyl]amino-naphthalene-1-sulfonic acid (I-AEDANS), 7-chloro-4 nitro-2,1,3-benzoxadiazole (NBD-Cl) and fluorescent maleimides. The surface of the membrane was labelled by reconstitution with fluorescent phospholipids (fluorescein and rhodamine derivatives). We found a distance of 4.1 nm from the nucleotide binding site to NBD (at Cys344), and the same distance to fluorescent maleimides (at Cys364). The AEDANS label (at Cys670,672) was found separated 3.5 nm from NBD, 4.4 nm from fluorescent maleimides, and 3.9 nm from the lipid matrix. The NBD label was 3.2 nm apart from fluorescent maleimides and 2.2 nm from the lipid matrix. Finally, fluorescent maleimides were found to be located 4.2 nm above the membrane surface. All these distances agree with a molecular model in which NBD is located in the stalk portion of the Ca(2+)-ATPase, near the surface of the membrane, and the rest of the probes are above it, in the globular domain of the protein. PMID- 8223617 TI - Molecular characterization of flavanone 3 beta-hydroxylases. Consensus sequence, comparison with related enzymes and the role of conserved histidine residues. AB - A heterologous cDNA probe from Petunia hybrida was used to isolate flavanone-3 beta-hydroxylase-encoding cDNA clones from carnation (Dianthus caryophyllus), china aster (Callistephus chinensis) and stock (Matthiola incana). The deduced protein sequences together with the known sequences of the enzyme from P. hybrida, barley (Hordeum vulgare) and snapdragon (Antirrhinum majus) enabled the determination of a consensus sequence which revealed an overall 84% similarity (53% identity) of flavanone 3 beta-hydroxylases from the different sources. Alignment with the sequences of other known enzymes of the same class and to related non-heme iron-(II) enzymes demonstrated the strict genetic conservation of 14 amino acids, in particular, of three histidines and an aspartic acid. The conservation of the histidine motifs provides strong support for the possible conservation of structurally similar iron-binding sites in these enzymes. The putative role of histidines as chelators of ferrous ions in the active site of flavanone 3 beta-hydroxylases was corroborated by diethyl-pyrocarbonate modification of the partially purified recombinant Petunia enzyme. PMID- 8223618 TI - Steady-state kinetics of substrate hydrolysis by vacuolar H(+)-pyrophosphatase. A simple three-state model. AB - The results of analyses of the steady-state kinetics of the vacuolar H(+) translocating pyrophosphatase (V-PPase) of native tonoplast vesicles isolated from etiolated hypocotyls of Vigna radiata (mung bean) and purified enzyme from the same source under a wide range of Mg2+, pyrophosphate (PPi) and K+ concentrations are consistent with a minimal reaction scheme in which dimagnesium pyrophosphate is the active substrate species and catalysis is mediated by preformed enzyme-Mg2+ complex. When account is taken of the sensitivity of the V PPase to ionic strength, additional kinetic interactions are not required to describe the behavior of the enzyme. N-Ethylmaleimide-protection assays show that the dissociation constant for Mg2+ binding in the absence of PPi is an order of magnitude smaller than that estimated from the steady-state kinetics of PPi hydrolysis. Two distinct Mg(2+)-binding sites are therefore invoked. Since the protective action of Mg2+ is independent of the nature of the monovalent cations and Mg2+ and K+ do not compete during substrate hydrolysis, divalent and monovalent cations are concluded to bind at separate sites. The pH dependencies of the kinetic parameters are consistent with the participation of groups of pKa 5.7 and 8.6 in substrate binding and groups of pKa 6.1 and 9.0 in the substrate conversion step, indicating that at least four ionizable groups are essential for catalysis. These findings are discussed with respect to the reaction mechanism of the V-PPase and the potential regulatory significance of cytosolic free Mg2+ and K+ in vivo. PMID- 8223619 TI - The ratio of protons translocated/hydride ion equivalent transferred by nicotinamide nucleotide transhydrogenase in chromatophores from Rhodospirillum rubrum. AB - The reduction of acetylpyridine adenine dinucleotide (AcPdAD+, an NAD+ analogue) by NADPH, in chromatophores treated with valinomycin, was accompanied by alkalinisation of the external medium, as measured by the absorbance change of added cresol red, a simple, non-binding pH indicator. Experiments with a stopped flow spectrophotometer showed that initial (linear) rates of alkalinisation persisted for 1-2s. From the results of experiments in which H+ uptake was driven by a series of short flashes of light, the dependence of the outward proton leak on the extent of H+ uptake was established. Thus, the proton leak was subtracted from the initial rate of alkalinisation during transhydrogenation to give the true proton-uptake rate. The correction factor was usually about 10%. The ratio of protons translocated/H- transferred from NADPH to AcPdAD+ (the H+/H- ratio) was 0.60 +/- 0.06. The transhydrogenation reaction between NAD+ and NADPH was measured in the presence of a regeneration system for NAD+ (pyruvate and lactate dehydrogenase). In addition to the accompanying proton-translocation reaction, scalar H+ consumption linked to the regeneration system was observed and permitted internal checks on the calibration of the cresol red absorbance changes. After correction for the proton leak and scalar proton uptake, an H+/H- ratio of 0.60 +/- 0.30 was calculated from the initial rates. The water-soluble polypeptide of transhydrogenase (Ths) was washed from a sample of chromatophores to inhibit transhydrogenation activity and the accompanying H+ uptake. Re addition of purified Ths to depleted chromatophores led to recovery of transhydrogenation activity and of H+ uptake. In this reconstituted system the H+/H- was similar to that in the native membranes. These results make it unlikely that the H+/H- ratio is artefactually low because chromatophores have a population of transhydrogenase which is not coupled to proton translocation. Further evidence that the mechanistic H+/H- ratio of chromatophore transhydrogenase is less than 1 was provided by an analysis of the kinetics of alkalinisation of the medium during reduction of AcPdAD+ by NADPH. It was shown that the progress of the transhydrogenation-induced alkalinisation was fitted by the sum of H+ uptake (the rate of transhydrogenation multiplied by the H+/H- ratio) plus the H+ leak, when the ratio was 0.6 but not when it was 1.0. The results are discussed in terms of the possible mechanism of energy coupling by transhydrogenase.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8223620 TI - Comparison of X-ray powder-diffraction data of various bacterial lipopolysaccharide structures with theoretical model conformations. AB - X-ray powder-diffraction experiments have been performed on dry samples of lipid A and various rough-mutant lipopolysaccharides (LPS) of Salmonella minnesota, Salmonella typhimurium and Escherichia coli. The diffraction patterns obtained indicated exclusively lamellar, bilayered arrangements in all samples. The periodicities were found to be in the range 4.5 nm for lipid A to 8.8 nm for Ra LPS. Upon treatment with water-saturated air, swelling of the lamellar structures was achieved, as indicated by shifts of reflections. The increase in bilayer dimensions normally was about 0.3 nm. X-ray intensities were used for the determination of the inner bilayer structure, i.e. for calculation of the one dimensional electron-density distribution across the bilayer. For lipid A and several Re-LPS, Rd2-LPS, Rd1-LPS and Rc-LPS samples, a striking coincidence of the electron-density distributions in the lipid-A domain was found, suggesting that in all these structures the lipid-A portion is similarly arranged. For Rb1 and Ra-LPS the lipid-A domain could not be resolved due to the limited number of observed reflections. For other Re-mutant lipopolysaccharide samples, quite different X-ray patterns were obtained. Some samples yielded diffraction patterns indicating a very high state of order in the lipid-A domain, whereas, in others, a significantly reduced order in the lipid-A domain was inferred. Comparison of the X-ray data with features of a calculated three-dimensional molecular model of lipopolysaccharide revealed reasonable agreement in molecular dimensions and bilayer structure. PMID- 8223621 TI - Modulation of tensin and vimentin expression in chick embryo developing cartilage and cultured differentiating chondrocytes. AB - It has been proposed that tensin, in association with several other proteins, mediates the micro-filament-integrin link. Here we describe the isolation of clones spanning about 5 kb from the 3' end of tensin mRNA from cultured chick embryo chondrocyte and embryonic heart cDNA libraries. Tensin expression was investigated in cultured chick embryo cells. It was observed that tensin expression is dependent upon substrate adhesion and it is turned off after 7 days of suspension culture. This process is reversible. Tensin expression is also regulated during cartilage cell differentiation in vivo; at Hamburger and Hamilton stage 39-40, non-hypertrophic tibial chondrocytes express both RNA and protein while hypertrophic chondrocytes do not. In the culture system the expression of vimentin, a major component of intermediate filaments, showed an opposite behaviour since the suspension culture enhances the accumulation of both vimentin and its mRNAs. Therefore in chick embryo cultured chondrocytes and in vivo, during cartilage development, cell shape changes and/or integrin extracellular matrix protein interactions may be involved in the regulation of these two genes coding for cytoskeletal proteins. PMID- 8223622 TI - Components of glycine reductase from Eubacterium acidaminophilum. Cloning, sequencing and identification of the genes for thioredoxin reductase, thioredoxin and selenoprotein PA. AB - The genes encoding thioredoxin reductase (trxB), thioredoxin (trxA), protein PA of glycine reductase (grdA) and the first 23 amino acids of the large subunit of protein PC of glycine reductase (grdC) belonging to the reductive deamination systems present in Eubacterium acidaminophilum were cloned and sequenced. The proteins were products of closely linked genes with 314 codons (thioredoxin reductase), 110 codons (thioredoxin), and 158 codons (protein PA). The protein previously called 'atypically small lipoamide dehydrogenase' or 'electron transferring flavoprotein' could now conclusively be identified as a thioredoxin reductase (subunit mass of 34781 Da) by the alignment with the enzyme of Escherichia coli showing the same typical order of the corresponding domains. The thioredoxin (molecular mass of 11742 Da) deviated considerably from the known consensus sequence, even in the most strongly conserved redox-active segment WCGPC that was now GCVPC. The selenocysteine of protein PA (molecular mass of 16609 Da) was encoded by TGA. The protein was highly similar to those of Clostridium purinolyticum and Clostridium sticklandii involved in glycine reductase. Thioredoxin reductase and thioredoxin of E. acidaminophilum could be successfully expressed in E. coli. PMID- 8223623 TI - Fusion of artificial membranes with mammalian spermatozoa. Specific involvement of the equatorial segment after acrosome reaction. AB - The fusogenic properties of bovine and human spermatozoa membranes were investigated, using phospholipid bilayers (liposomes) as target membranes. Fusion was monitored by following lipid mixing, as revealed by an assay based on resonance-energy transfer. In addition, fusion was visualized by fluorescence microscopy, using fluorescent lipid vesicles. Cryopreserved bovine sperm fused with liposomes before induction of the acrosome reaction, fluorescence being located in essentially all spermatozoa membrane domains. Fresh bovine and human spermatozoa fused with liposomes only after the induction of the acrosome reaction, as triggered by calcium ionophore A23187 or zonae pellucidae (proteins), while the fluorescence distribution was mainly restricted to the equatorial segment (ES). However, with spermatozoa that had undergone a freeze/thawing cycle, domains other than ES also became labeled. Hence, the redistribution of the lipid probes over the entire membrane occurring during lipid mixing with cryopreserved bovine sperm is probably related to membrane perturbations caused by long-term cryopreservation. Fusion with liposomes was governed by spermatozoa factors and required the presence of acidic phospholipids like cardiolipin and phosphatidylserine in the liposomal bilayer. Incorporation of the zwitterionic lipid phosphatidylcholine in the vesicles inhibited the fusion reaction. Fusion was pH dependent. The results indicate that the ES is the primary domain of spermatozoa membranes that harbours the fusogenic capacity of sperm. Liposomes appear a valuable tool in further characterizing the properties of this domain, which has been claimed [Yanagimachi, R. (1988) in The physiology of reproduction (Knobil, E. & Neill, J., eds) pp. 135-185, Raven Press, New York] to represent the putative, initial fusion site for the oocyte. PMID- 8223624 TI - Thermodynamics of the maleate and citraconate hydration reactions catalysed by malease from Pseudomonas pseudoalcaligenes. AB - Malease from Pseudomonas pseudoalcaligenes catalyses the hydration of both maleate and citraconate to D-malate and D-citramalate, respectively. The Kapp for these hydration reactions were 2050 and 104, respectively, under standard biochemical conditions (25 degrees C, pH 7.0, I = 0.1). The influence of the pH (6.0-8.5) on Kapp was determined. The Gibbs-free-energy changes under standard biochemical conditions for the hydration of the dianionic acids were calculated to be -19.28 kJ.mol-1 and -11.65 kJ.mol-1, respectively. From the obtained data together with data from the literature, the Gibbs free energy of formation of maleate2- and citraconate2- were calculated to be -588.91 kJ.mol-1 and -600.56 kJ.mol-1, respectively. The influence of the temperature (10-40 degrees C) on Kapp was determined for both hydration reactions. The enthalpy change (delta H degrees') and entropy change (delta S degrees') under standard biochemical conditions for the maleate2- (delta H degrees' = 18.07 kJ.mol-1, delta S degrees' = 2.94 J.mol-1 x K-1) and citraconate2- (delta H degrees' = -22.55 kJ.mol-1, delta S degrees' = -35.92 kJ.mol-1 x K-1) hydration reactions were calculated. The reaction rate of malease from Ps. pseudoalcaligenes was studied for both hydration reactions as a function of temperature. From these studies, the Gibbs free energies of activation for the maleate and citraconate hydration reactions catalysed by malease from Ps. pseudoalcaligenes were calculated to be 62.21 kJ.mol-1 and 63.43 kJ.mol-1, respectively. PMID- 8223625 TI - Characterization of the KdpD protein, the sensor kinase of the K(+)-translocating Kdp system of Escherichia coli. AB - KdpD and KdpE, proteins that control expression of the kdpFABC operon, are members of the class of sensor kinase/response regulator proteins. Using polyclonal antibodies raised against the KdpD protein, we have been able to identify and to localize the chromosome-encoded KdpD protein in the cytoplasmic membrane of Escherichia coli. Furthermore, it has been possible to detect differences in the expression of the KdpD protein according to the K+ concentration in the growth medium. The phosphorylation capacity of the plasmid encoded KdpD protein and the phospho-transfer to KdpE was investigated. We found that both reactions were strictly dependent on the ionic conditions of the assay medium. Based on optimized conditions, we were able to detect phosphorylation of the chromosome-encoded KdpD protein. Furthermore, replacement of the conserved histidine (His673), the predicted phosphorylation site in KdpD, by glutamine revealed that phosphorylation of KdpD was no longer possible. PMID- 8223626 TI - Comparison of the biochemical properties of unprocessed and processed forms of the small GTP-binding protein, rab6p. AB - The rab6 protein (rab6p) belongs to a large family of ras-like low-molecular-mass GTP-binding proteins thought to be involved in the regulation of intracellular transport in mammalian cells. When expressed in the baculovirus/insect cell system, two major forms of rab6p are obtained; a 24-kDa cytosolic unprocessed form and a 23-kDa membrane-bound form which represents the processed lipid modified protein. Here, we have purified both forms to homogeneity and we have studied and compared their biochemical properties. Unprocessed and processed rab6p display similar binding-rate constants (kon) for GDP and GTP (1-1.9 microM 1 min-1). However, significant differences exist in the dissociation constants of bound guanine nucleotides. Processed rab6p in low and high magnesium solutions displays similar koff values for GTP and GDP. However, unprocessed rab6p has a koff value higher for GDP than for GTP in both low and high magnesium solutions. Their intrinsic GTPase activities also differ; unprocessed rab6p has an almost undetectable GTPase activity, whereas that of processed rab6p is in the same range as that reported for other ras and ras-like GTP-binding proteins (0.012 +/- 0.002 min-1). These results suggest that post-translational modifications of rab6p might induce subtle changes in the three-dimensional structure of the protein which affect the guanine-nucleotide-binding/hydrolysis activity. PMID- 8223627 TI - Partial purification and properties of enzymes involved in the processing of a chloroplast import protein from Chlamydomonas reinhardii. AB - Two stromal peptidases (SPP-1 and SPP-2) were partially purified from chloroplasts of Chlamydomonas reinhardii. They specifically processed in vitro the precursor of the small subunit of ribulose-1,5-bisphosphate carboxylase (pSS), which had been synthesized by using the cloned rbcS-2 gene of Chlamydomonas. SPP-1 shortened pSS to an intermediate-sized form (iSS), while SPP 2 cut pSS and iSS to the mature small subunit SS. N-terminal amino acid sequencing demonstrated that the reaction product obtained with SPP-2 had an N terminus identical to natural SS, and that iSS derived from pSS by hydrolysis at the amino side of the methionine located within the transit sequence. By gel filtration, apparent molecular masses of 340 kDa and 90 kDa were determined for SPP-1 and SPP-2, respectively. The comparison of these molecular masses with the protein patterns obtained by SDS/PAGE of the partially purified enzymes suggested that at least SPP-1 was a multimeric protein. The enzymes differed also in their pH optima of about 8 (SPP-1) and 9 (SPP-2) and in their sensitivity to different inhibitors. However, both enzymes seem to be serine proteases as they were completely blocked by N-alpha-tosyl-L-lysinechloromethane or tosylphenylalaninechloromethane, respectively. Competition experiments, using either mature SS or a synthetic hexadecapeptide with 15 amino acids similar to the C-terminal end of the transit sequence of pSS, indicated that SPP-2 had some affinities not only to the transit sequence of pSS, but especially to sequences in the mature protein part. We conclude that SPP-2 in Chlamydomonas is the enzyme involved in import of pSS into chloroplasts and responsible for its processing by a one-step mechanism. PMID- 8223628 TI - The putative zinc-finger protein WZF1 interacts with a cis-acting element of wheat histone genes. AB - A nonamer motif (CATCCAACG) that is one of the cis-acting elements identified in the proximal promoter region of some wheat histone genes is included in the region that interacts with the wheat DNA-binding protein, HBP (histone gene binding protein)-2. To obtain structural and functional information about this DNA-binding protein, we attempted to isolate a cDNA clone encoding HBP-2 on the basis of its ability to bind to a nonamer-containing 38-bp DNA fragment. Southwestern screening of a wheat cDNA library with concatenated 38-residue oligonucleotides as the probe produced one candidate clone. Nucleotide sequence analyses of this cDNA clone and the corresponding genomic clone showed that the protein deduced from the nucleotide sequence consisted of 261 amino acids and contained a set of zinc-finger motifs similar to those found in many eukaryotic transcription factors. The protein, named WZF1 (wheat zinc-finger protein 1), which was expressed from the cDNA in Escherichia coli cells, bound specifically and metal-ion-dependently to the nonamer-containing oligonucleotide. The WZF1 mRNA was highly expressed in the root apexes of wheat seedlings, but less so in the proximal portion of young leaves; whereas, histone H3 mRNA was highly expressed in both tissues. The expression patterns of the WZF1 and histone H3 genes in the early stages of germination differed, expression of the WZF1 gene being almost constant but not that of the H3 gene. The relationship of WZF1 and HBP-2 and the possible role of WZF1 in the histone gene expression were discussed. PMID- 8223629 TI - A new brain-specific 14-kDa protein is a phosphoprotein. Its complete amino acid sequence and evidence for phosphorylation. AB - We previously reported a new brain-specific protein with a molecular mass of 14 kDa, specifically present in synapses around neurons but not in glial cells [Nakajo, S., Omata, K., Aiuchi, T., Shibayama, T., Okahashi, I., Ochiai, H., Nakai, Y., Nakaya, K. & Nakamura, Y. (1990) J. Neurochem. 55, 2031-2038]. In the present study, we determined the primary structure of this protein, found that it is phosphorylated in vitro and in vivo, and designated it phosphoneuroprotein 14 (PNP 14). The protein is a single polypeptide with 134 amino acid residues (molecular mass = 14122 Da), and it contains a hydrophobic region at the center of the molecule. The carboxy-terminal region has all seven proline residues, and is rich in glutamic acid, which contribute to the acidic property of the protein. The amino-terminal region possesses four unique repetitive motifs, Glu(Ser)-Lys Thr-Lys-Glu(Gln)-Gly(Gln)-Val(Ala). When a cytosolic fraction prepared from rat cerebral cortex was incubated with [gamma-32P]ATP, 32P was incorporated into PNP 14. Phosphorylated PNP 14 was immunoprecipitated from rat brain synaptosomes labeled metabolically with [32P]orthophosphate. Injection of [32P]orthophosphate into the third ventricle of rat brain resulted in incorporation of radioactive phosphate into PNP 14. We have also found that Ca2+, calmodulin-dependent protein kinase II phosphorylates serine residue(s) of PNP 14 in vitro. The results suggest that PNP 14 may be important to neuronal cells, but not to glial cells, and that its physiological functions may be controlled by the phosphorylation reaction. PMID- 8223630 TI - Structural and functional properties of a multi-enzyme complex from spinach chloroplasts. 1. Stoichiometry of the polypeptide chains. AB - Antibodies have been raised specifically against chloroplast phosphoribulokinase, glyceraldehyde-3-phosphate dehydrogenase and ribulose 1,5-bisphosphate carboxylase-oxygenase. Each of these antibodies recognizes the same macromolecular entity isolated and purified from chloroplasts. This entity is a multi-enzyme complex, previously isolated and made up of ribose-phosphate isomerase, phosphoribulokinase, ribulose 1,5-bisphosphate carboxylase-oxygenase, phosphoglycerate kinase and glyceraldehyde-3-phosphate dehydrogenase. Under denaturing conditions the multi-enzyme complex contains two polypeptides of 54 kDa and 15 kDa corresponding to the large and the small subunits of ribulose 1,5 bisphosphate carboxylase-oxygenase, the two polypeptides of the glyceraldehyde-3 phosphate dehydrogenase of 39 kDa and 37 kDa, one polypeptide of 40 kDa pertaining to phosphoribulokinase and one polypeptide of 30 kDa very likely pertaining to ribose-phosphate isomerase. The combined use of immunochemical and densitometric techniques allows one to determine the number and the stoichiometry of the various types of polypeptide chains and to compare them with the quaternary structure of the corresponding isolated enzymes. Ribulose 1,5 bisphosphate carboxylase-oxygenase of higher plants consists of eight large and eight small subunits. Glyceraldehyde-3-phosphate dehydrogenase is made up of two types of polypeptide chains called A and B and its simplest quaternary structure is A2B2. Finally, phosphoribulokinase is a dimer made up of two identical subunits. Therefore, for the three isolated enzymes, the stoichiometry of the polypeptide chains is always 1:1. Within this multi-enzyme complex, there are two subunits of phosphoribulokinase, two A and B subunits of glyceraldehyde-3 phosphate dehydrogenase and two large and four small subunits of ribulose 1,5 bisphosphate carboxylase-oxygenase. Therefore the number and the stoichiometry of the polypeptide chains of phosphoribulokinase and glyceraldehyde-3-phosphate dehydrogenase are the same in the multi-enzyme complex and in the free enzymes, but those of ribulose 1,5-bisphosphate carboxylase-oxygenase are completely different. This conclusion that the multi-enzyme complex contains two active sites for ribulose 1,5-bisphosphate may be confirmed independently by kinetic inhibition studies using 6-phosphogluconate. PMID- 8223631 TI - Structural and functional properties of a multi-enzyme complex from spinach chloroplasts. 2. Modulation of the kinetic properties of enzymes in the aggregated state. AB - The carboxylase activity of free ribulose 1,5-bisphosphate carboxylase-oxygenase has been compared to that of the five-enzyme complex present in chloroplasts. Kinetic results have shown that the V/active site is lower for the free enzyme than for the complex. Conversely the Km is smaller for the complex than for the free enzyme. This implies that the catalytic activity of the enzyme is enhanced when it is embedded in the complex. Under reducing conditions and in the presence of reduced thioredoxin, inactive oxidized phosphoribulokinase, free in solution or inserted in the multi-enzyme complex, becomes active. The kinetics of this activation process has been studied and shown to be exponential. The time constant of this exponential decreases, for the free enzyme, as thioredoxin concentration is increased. Alternatively, for the enzyme embedded in the complex, this time constant increases with thioredoxin concentration almost in a linear fashion. This implies that the complex is much more rapidly activated by reduced thioredoxin than is the free phosphoribulokinase. The variation of the amplitude of this activation process as a function of thioredoxin concentration is a hyperbola. The concentration of thioredoxin which results in half the asymptotic value of this hyperbola is smaller for the complex than for the free enzyme. A kinetic model has been proposed and the dynamic equations resulting from this model have been derived. They fit the experimental results exactly. From the variation of the amplitude of the activation process one may derive the binding constants of thioredoxin on either the oxidized enzyme or on a partly dithiothreitol-reduced enzyme (both of them free or inserted in the complex). In either case, the affinity of reduced thioredoxin is larger for the complex than for the free enzyme. The individual values of some of the rate constants have also been estimated from the variation of the time constants as a function of thioredoxin concentration. Taken together, these results show that at least two enzymes, ribulose 1,5-bisphosphate carboxylase-oxygenase and phosphoribulokinase, have quite different kinetic properties depending on whether they are in free solution or embedded in the multi-enzyme complex. PMID- 8223632 TI - Isolation and characterization of cDNAs from Atlantic cod encoding two different forms of trypsinogen. AB - The cDNAs encoding two different anionic forms of Atlantic cod trypsinogen have been isolated and sequenced. The nucleotide sequences include the 5'-noncoding and 3'-noncoding regions in addition to preproenzymes of 241 amino acids. These consist of hydrophobic signal peptides, activation hexapeptides and trypsins of 222 amino acid residues. The cod trypsins contain all the major structural features common to trypsins such as the catalytic triad His57, Asp102 and Ser195. Furthermore, the obligatory Asp189 and the six disulphide bonds are conserved. Eight amino acid residues are different between the isozymes, leading to a difference of four charges. Both cod trypsins are one-amino-acid-residue shorter than most mammalian trypsins as a result of deletion of proline at position 152, and have a high methionine content. In addition, the cod preproenzyme signal and activation peptides differ markedly from their mammalian analogues. The amino acid identity between the cod and bovine trypsins is approximately 60%. PMID- 8223633 TI - Expression of human cytochrome c oxidase subunits during fetal development. AB - Expression of human cytochrome c oxidase (COX) subunits was examined at fetal (20 28 weeks) and adult state by Northern blot hybridization with mRNA from liver, heart, skeletal muscle, and intestine. The data were related to COX and citrate synthase activities and to immunodetected COX subunits (II/III, IV, VIIaH). In liver little changes of COX transcripts are observed from fetal to adult state. In contrast, in heart and skeletal muscle all transcripts of COX subunits increase between 2-20-fold, when related to the amount of 28S rRNA. In fetal heart and skeletal muscle the relative amounts of the liver-type transcripts of subunit VIa were 30% and 25% from total VIa transcripts (VIaL+VIaH), respectively, but decrease to only 2-5% at adult state. The liver-type transcripts of subunit VIIa occur to 50% in fetal heart and skeletal muscle, which remained unchanged in adult heart and decrease to 5-8% in adult skeletal muscle. The results clearly indicate a switch of gene expression in heart and skeletal muscle during development, from the liver type to the heart/muscle type of subunit VIa (and partly VIIa). PMID- 8223634 TI - Mutational analysis of phospholipase C-beta 2. Identification of regions required for membrane association and stimulation by guanine-nucleotide-binding protein beta gamma subunits. AB - Members of the beta isozyme subfamily of the phosphoinositide-specific phospholipases C (PLC beta) have recently been shown to be stimulated by both guanine-nucleotide-binding protein alpha and beta gamma subunits. The alpha subunits of the Gq class activate PLC beta isozymes in the order of PLC beta 1 > or = PLC beta 3 >> PLC beta 2, which is different from the order of PLC beta 3 > PLC beta 2 > PLC beta 1 for beta gamma subunit stimulation. The C-terminal region of PLC beta 1, in particular the sequence between Thr903 and Leu1142, has been shown to be involved in interacting with activated alpha q subunits and to contain a region required for efficient membrane association of PLC beta 1 [Park, D., Jhon, D.-Y., Lee, C.-W., Ryu, S. H. & Rhee, S. G. (1993) J. Biol. Chem. 268, 3710-3714, and Wu, D., Jiang, H., Katz, A. & Simon, M. I. (1993) J. Biol. Chem. 268, 3704-3709]. To examine the structure-function relationships of a PLC beta isozyme highly sensitive to beta gamma subunit stimulation, we have altered the cDNA of PLC beta 2 by site-directed mutagenesis and have examined the effects of these structural alterations on the functional properties of the mutant polypeptides. The results show that the C-terminal region of PLC beta 2 downstream of Phe818, which corresponds to Tyr816 of PLC beta 1, contains a region essential for membrane association, but is required neither for the interaction of PLC beta 2 with Ca2+ and the phospholipid substrate, nor for beta gamma subunit stimulation of PLC beta 2. These data suggest that PLC beta isozymes are activated by alpha q and beta gamma subunits via distinct domains. PMID- 8223635 TI - Biosynthesis of vitamin B12 in anaerobic bacteria. Transformation of 5 hydroxybenzimidazole and 5-hydroxy-6-methylbenzimidazole into 5,6 dimethylbenzimidazole in Eubacterium limosum. AB - Eubacterium limosum transformed [2-13C]5-hydroxybenzimidazole not only into [2 13C]5-hydroxybenzimidazolylcobamide, but also into [2-13C]5-methoxy-6 methylbenzimidazolylcobamide and into [2-13C]5,6-dimethylbenzimidazolylcobamide (vitamin B12). [2-13C]5-Hydroxy-6-methyl-benzimidazole was used by this bacterium to form [2-13C]5-hydroxy-6-methylbenzimidazolyl-cobamide, [2-13C]5-methoxy-6 methylbenzimidazolylcobamide and [2-13C]5,6-dimethylbenzimidazolylcobamide. The 1H-NMR spectrum of the 5,6-dimethylbenzimidazole isolated from the 13C-labeled vitamin B12 preparations revealed that the externally added bases had been transformed into the vitamin B12 base almost without dilution of the label. This suggests that 5-hydroxybenzimidazole and 5-hydroxy-6-methylbenzimidazole are precursors of 5,6-dimethylbenzimidazole. On the basis of these results, a hypothetical scheme for the biosynthesis of 5,6-dimethylbenzimidazole via 5 hydroxybenzimidazole and 5-hydroxy-6-methylbenzimidazole is discussed. This scheme can also explain the formation of the other benzimidazole bases found in natural vitamin B12 analogs. PMID- 8223636 TI - Broad binding-site specificity and affinity properties of octamer 1 and brain octamer-binding proteins. AB - The ubiquitous Pit-1-Oct-1-Unc-1 (POU)-domain protein octamer 1 (Oct-1) has been observed to bind specifically to a number of degenerate and dissimilar sequences. We have used antibodies directed against a C-terminal Oct-1 peptide to immunoselect binding sequences for HeLa cell Oct-1 from random-sequence oligonucleotides and we describe the isolation of binding sequences of considerable heterogeneity. Although our consensus alignment indicated a 9-bp TATGCAAAT motif with AT-rich flanking sequences, this binding motif is not immediately obvious in the population of sequences and no clone actually contained this sequence. Screening these Oct-1-binding sequences with a mouse whole-brain extract demonstrated that the neuronal octamer-binding proteins exhibit similar but distinct DNA sequence specificities. Unlike the reported selection of binding sequences for other transcription factors, the dependence of Oct-1-binding affinity upon sequence did not correspond tightly to the degree of conservation at particular positions of the consensus sequence. Our results suggest that either base-specific hydrogen bonding is not the only major determinant of binding affinity and specificity, or that Oct-1 binding to some sequences is mechanistically different from its binding to an octamer. These results exemplify the potential to overlook binding sites for some factors by searching gene sequences with a consensus nucleotide sequence. PMID- 8223637 TI - The promoter region of the human type-I-DNA-topoisomerase gene. Protein-binding sites and sequences involved in transcriptional regulation. AB - We examined the promoter of the human type-I-DNA topoisomerase gene (hTOP1) for regions protected against DNase I digestion by nuclear proteins from HeLa or from adenovirus-transformed 293 cells. We identified ten protected DNA sequences within 580 bp of DNA upstream of the transcriptional-start sites and one additional site, which is located between the two clusters of transcriptional start sites. Several of these protein-binding sites have significant similarities to recognition sequences of known transcription factors including factors Sp1, octamer transcription factor, cAMP-responsive-element-binding protein (CREB/ATF), NF-kappa B and members of the Myc-related family of basic/helix-loop helix/leucine-zipper proteins. Other protein-binding sites show less or no similarities to known consensus sequences. We investigated the physiological significance of these protein-binding sites using a set of deletion and nucleotide-exchange mutants. We conclude that the expression of the hTOP1 gene is regulated by a complex network of negatively and positively acting transcription factors. PMID- 8223638 TI - Characterization and purification of the solubilized pituitary adenylate-cyclase activating polypeptide-1 receptor from porcine brain using a biotinylated ligand. AB - A specific receptor for the brain-gut neuropeptide pituitary adenylate-cyclase activating polypeptide (PACAP-1 receptor) was solubilized with Chapso from porcine brain plasma membranes and purified. Binding of 125I-PACAP(1-27) to the solubilized material was reversed equipotently by unlabeled PACAP(1-27) and PACAP(1-38). Soluble receptors retained the binding affinities and specificities of the plasma membrane fraction. Scatchard analysis of equilibrium-binding data indicated the existence of a single high-affinity binding site (Kd = 0.23 nM, Bmax = 1.2 pmol/mg protein). Binding of 125I-PACAP(1-27) to solubilized receptors was not affected by guanosine nucleotides, suggesting that solubilization dissociates the PACAP-1 receptor/guanosine-nucleotide-binding protein complex. Affinity cross-linking of 125I-PACAP(1-27) to soluble PACAP-1 receptors identified a specifically labeled 60-kDa protein. Enzymic deglycosylation of soluble affinity-labeled receptors reduced the apparent molecular mass by 10 kDa. The solubilized receptor glycoprotein was purified 4-5-fold by lectin-adsorption chromatography on wheatgerm agglutinin immobilized on agarose. S-Biotinyl[Ala28 34, Cys35]PACAP(1-35) was synthesized, immobilized on streptavidin-coated magnetic Sepharose beads and used to further affinity-purify wheatgerm-agglutinin eluted receptor material. This more than 6000-fold enriched PACAP-1-receptor preparation retained single-class high-affinity binding and consisted of an almost homogenous 55-60-kDa protein identified by silver staining. In conclusion, we established a rapid method for purification of PACAP-1 receptors, allowing further studies to be performed by protein chemistry. PMID- 8223639 TI - Higher plant mitochondria encode an homologue of the nuclear-encoded 30-kDa subunit of bovine mitochondrial complex I. AB - We describe the structure and expression of a wheat mitochondrial gene, which codes for a subunit of mitochondrial NADH dehydrogenase. The deduced protein sequence has 70% similarity to the 30-kDa subunit of bovine mitochondrial complex I and 65% similarity to the 31-kDa subunit of Neurospora crassa complex I, components of the iron-sulfur-protein fraction, both nuclear-encoded proteins. We named this wheat mitochondrial gene as nad9. The wheat nad9 gene is transcribed in a single mRNA of 0.9 kb that is edited (C-to-U conversions) in 14 positions. Transcript mapping revealed that the first ATG codon is just 20 nucleotides downstream of the mRNA 5' end and that the 3' end is just 23 nucleotides downstream of the nad9 stop codon. The expression of the nad9 gene in plant mitochondria was studied. Polyclonal antibodies prepared against a wheat NAD9 fusion protein specifically recognise the 30-kDa subunit of bovine mitochondrial complex I and a 27.5-kDa protein in the membrane fractions of wheat, maize and common bean mitochondria, whereas the same serum recognizes a 30-kDa protein in the mitochondria of pea, chickpea and lentil. PMID- 8223640 TI - Reconstruction of translation. Evidence for the involvement of the rescue protein in the association/dissociation of ribosomal subunits. AB - The in vitro reversal of conditionally lethal mutations has greatly aided the study of translation. N4316 is a mutant of Escherichia coli that has a temperature-sensitive defect in a protein called the rescue protein. Without the rescue protein, translation in vivo and in vitro is drastically reduced and frameshift errors, as well as increased read-through of nonsense codons, occurs. Using reversal of temperature-sensitivity as an assay, the rescue protein was purified from a ribosomal eluate of the parental (D10) strain. Composite polyacrylamide/agarose gel electrophoresis and sedimentation on sucrose density gradients were employed to examine the distribution of 70S ribosomes and ribosomal subunits in the mutant (N4316) and the parental (D10) extracts at restrictive (43 degrees C) and non-restrictive (35 degrees C) temperatures. Fewer polysomes and a larger proportion of 70S ribosomes relative to subunits were observed at 43 degrees C with N4316, but not with D10 extracts. Addition of the rescue protein had no effect at 35 degrees C with either strain, but restored the polysome pattern of N4316 at 43 degrees C. The purified rescue protein labelled by methylation retained activity and bound preferentially to 30S subunits. Rescue bound to 30S particles prevented the action of IF-3 fostering formation of 70S ribosomes. Thus the rescue protein enables formation of 70S ribosomes from 30S and 50S subunits. 70S ribosomes which contain the rescue protein are active in translation and resist dissociation induced by high centrifugal fields. We propose that the rescue protein alters the conformation of 70S ribosomes resulting in a tighter association of subunits which, in turn, fosters both higher rates and increased accuracy of translation. PMID- 8223641 TI - Conformations and conformational changes of four Phe-->Trp variants of the DNA binding histone-like protein, HBsu, from Bacillus subtilis studied by circular dichroism and fluorescence spectroscopy. AB - Circular dichroic spectra in the region 180-260 nm of the DNA-binding histone like protein, HBsu, from Bacillus subtilis and of four mutants with a Phe residue replaced by Trp, i.e. [F29W]HBsu, [F47W]HBsu, [F50W]HBsu and [F79W]HBsu, show minor differences only and demonstrate the general similarity of the conformations of these proteins. Fluorescence maxima at 315-320 nm and 330-335 nm indicate a more hydrophobic environment or a more effective stacking of Trp residues in mutants [F29W]HBsu and [F50W]HBsu in comparison to [F47W]HBsu and [F79W]HBsu, respectively. Unfolding of the mutants in high-ionic-strength buffers by increasing concentrations of urea results in a red shift of the fluorescence emission maxima to about 350 nm; the fluorescence intensities decrease strongly for [F29W]HBsu and [F50W]HBsu but show a small increase for [F47W]HBsu and [F79W]HBsu. The data suggest complex unfolding patterns with subtle differences between the single mutants. The circular dichroic spectra in the region 250-320 nm are dominated by the effects of the Trp residues and signal position-dependent differences in the environment of the Trp residues. The conformations of the mutant proteins depend on the ionic strength of the buffer and become more stable against unfolding by denaturants or increasing temperatures at higher ionic strength. At low ionic strength a pronounced protein-concentration dependence of the conformation of the mutants is seen. PMID- 8223642 TI - Molecular cloning, expression and functional characterization of rabbit anticoagulant vitamin-K-dependent protein S. AB - Vitamin-K-dependent protein S is an anticoagulant plasma protein which functions as cofactor to activated protein C (APC) in the degradation of coagulation factors Va and VIIIa. In addition, it interacts with C4b-binding protein (C4BP), a regulator of the complement system. Using a human protein S cDNA clone as probe, cDNA clones for rabbit protein S were isolated from a rabbit liver cDNA library. The cDNA sequence encoded the mature protein S and 12 residues of the leader sequence. The amino acid sequence of the single-chain 634-amino-acid residue-long rabbit protein S molecule was 82% and 81% identical to those of human and bovine protein S, respectively. Northern blotting demonstrated protein S mRNA not only in liver but also in reproductive organs (testis, ovary and uterus), in lung and brain. Recombinant rabbit protein S was expressed in eucaryotic cells and found to be post-translationally modified, i.e. it had the correct amino terminus, contained N-linked carbohydrate side chains, gamma carboxyglutamic acid residues and beta-hydroxylated aspartic acid and asparagine residues. Recombinant rabbit protein S bound calcium like its human counterpart, as judged by its migration in the presence of calcium on agarose-gel electrophoresis. Rabbit protein S has been reported to be species specific with respect to its interaction with APC and not to function with bovine APC. However, we found it to act as cofactor to both human and bovine APC, albeit it was somewhat more efficient with human than with bovine APC. Rabbit protein S, like its human and bovine counterparts, bound human C4BP in a reaction which was associated with the loss of its APC-cofactor activity. However, unlike human plasma, rabbit plasma appeared to contain only the free form of protein S as a radiolabeled rabbit protein S tracer added to rabbit plasma migrated as free protein S on agarose-gel electrophoresis. Addition of human C4BP to rabbit plasma resulted in the formation of a C4BP-protein-S complex, suggesting an explanation for the absence of complexed protein S in rabbit plasma to be sought for in the structure of rabbit C4BP. PMID- 8223643 TI - Sites of nidogen cleavage by proteases involved in tissue homeostasis and remodelling. AB - The cleavage of recombinant mouse nidogen in its native form was examined with granule-stored proteases (leucocyte elastase, mast-cell chymase), blood proteases (thrombin, plasmin, kallikrein), matrix metalloproteinases (stromelysin, matrilysin, collagenases) and, for comparison, with trypsin and the endoproteinase Glu-C. More than 50 major cleavage sites were identified by Edman degradation of several large fragments and smaller peptides. The data show an almost exclusive localization of protease-sensitive sites to the flexible segment, connecting the N-terminal globular domains G1 and G2, and within the C terminal, laminin-binding domain G3. Domains G1, G2 and the rod-like segment were much more stable against proteolysis. Kinetic analysis indicated a fast cleavage of several different sites in the link region followed by destruction of G3 but this was to some extent variable depending on the particular protease. Leucocyte elastase was identified as the most active protease in the cleavage of nidogen whilst stromelysin, matrilysin, plasmin and kallikrein were of distinctly lower activity. No cleavage could be detected with interstitial collagenase and gelatinase A. The peptide analyses also allowed the location of two disulfide bridges within the G3 domain. Complex formation between nidogen and laminin fragments caused some protection against cleavage by thrombin, leucocyte elastase and stromelysin particularly in domain G3. The data indicate a relatively uniform cleavage pattern of nidogen which may be relevant in the context of protein/ligand-binding activities associated with domains G2 and G3. The proteolytic processes involved in remodelling and the cellular penetration of basement membranes could therefore be essential for the modulation of the mediator function of nidogen. PMID- 8223644 TI - Four 9-kDa proteins excreted by somatic embryos of grapevine are isoforms of lipid-transfer proteins. AB - Four 9-kDa small extracellular proteins produced by embryogenic cultures in the absence of auxin have been purified from the extracellular medium of grapevine somatic embryo cultures through cation-exchange chromatography and hydrophobic interaction chromatography. The partial amino-acid sequences reflect high similarities between the four proteins as well as with the sequences established for carrot, spinach, millet and maize nonspecific lipid-transfer proteins. All these sequences show conservation of three cysteines at positions 4, 14 and 30 32, as well as glycine, valine, tyrosine and lysine residues at positions 5, 7, 17 and 37, respectively. In-vitro lipid-transfer assays reveal that the four proteins catalyze the transfer of phosphatidylcholine from liposomes towards mitochondria with an efficiency similar or higher than that of a purified maize lipid-transfer protein. PMID- 8223645 TI - Disulfide interchange reactions in 11S globulin subunits of Cruciferae seeds. Relationships to gene families. AB - Cruciferin, the main storage protein in rapeseed (Brassica napus L.), is a legumin-like 11S globulin. Using SDS/PAGE cruciferin was shown to be composed of different subunits consisting of alpha S and beta S polypeptides, which were disulfide linked, and also closely related free alpha f and beta f polypeptides which were not covalently linked. The origin of these latter free chains was found to be the result of disulfide interchange reactions occurring during SDS induced dissociation of the hexameric 11S complex. These reactions involved the rupture of the alpha S-beta S interchain disulfide link and, as shown by specific thiol fluorescent labeling, the subsequent formation of an intra-beta-chain disulfide bond. N-terminal amino acid sequence determinations revealed that these reactions were related to particular cruciferin subunit beta chains having an additional cysteine residue (position 11) near the residue (position 7) implicated in the inter-chain disulfide bridges. Such a sequence could explain the origin of the free and linked polypeptides as also shown in SDS/PAGE analysis of radish (Raphanus sativum L.) and Arabidopsis thaliana H. cruciferins. These two polypeptide classes may be considered to represent phenotypic biochemical markers of two different Cruciferae globulin gene families. PMID- 8223646 TI - Isolation, structural characterization and biological function of a lysine conopressin in the central nervous system of the pharyngobdellid leech Erpobdella octoculata. AB - Several neuropeptides are suspected to act on the control of hydric balance in leeches. One of these peptides, a peptide immunoreactive to an antibody against oxytocin, was previously characterized from the central nervous system of the leech Erpobdella octoculata [Salzet, M., Wattez, C., Verger-Bocquet, M., Beauvillain, J.-C. & Malecha, J. (1993) Brain Res. 601, 173-184]. This paper reports the isolation from the central nervous system of E. octoculata of another peptide of the oxytocin/vasopressin family; a lysine-vasopressin-like molecule. Its purification was performed by reverse-phase high-performance liquid chromatography combined with both dot immunobinding assay and enzyme-linked immunosorbent assay for lysine-vasopressin. The amino acid sequence was established by Edman degradation and confirmed by electrospray-mass-spectrometry measurement. The nonapeptide obtained corresponded to the lysine-conopressin previously isolated from the venom of the mollusc Conus geographus [Cruz, L. L., de Santos, V., Zafaralla, G. C., Ramilo, C. A., Zeikus, R., Gray, W. R. & Olivera, B. M. (1987) J. Biol. Chem. 262, 15821-15824]. In leeches, synthetic lysine-conopressin exerts a diuretic effect which can be compared to that of the arginine-vasopressin-like peptide isolated in the Insect Locusta migratoria [Proux, J., Miller, C. A., Li, J. P., Carney, R. L., Girardie, A., Delaage, M. & Schooley, D. A. (1987) Biochem. Biophys. Res. Commun. 149, 180-186]. PMID- 8223647 TI - Structural requirements for processing of pro-adipokinetic hormone I. AB - We found that a seven-residue sequence in pro-adipokinetic hormone I (proAKH I) which precedes the endopeptidase cleavage site is predicted to form an omega loop. Molecular modelling experiments indicated that a stable omega loop may form at this site, and suggested that loop stability may depend on the C-terminal loop residue, Lys12. The importance of this residue in proAKH I processing was confirmed by the observation that replacement of Lys12 by thialysine, a Lys analog with an altered side chain, prevented processing in vivo. In addition we showed by molecular modelling that this side-chain alteration may prevent formation of an omega loop. Together, these approaches lead us to propose that an omega loop may serve as a recognition motif in proAKH I processing. PMID- 8223648 TI - Expression of human interferon omega 1 in Sf9 cells. No evidence for complex-type N-linked glycosylation or sialylation. AB - Human interferon omega 1 (IFN-omega 1) was expressed in Spodoptera frugiperda Sf9 insect cells using the baculovirus expression system. Half of the protein purified by immunoaffinity chromatography was shown to be N-glycosylated at the same site as the natural IFN-omega 1. The degree of glycosylation was independent of the expression rate. While natural IFN-omega 1 was shown to carry complex-type oligosaccharides [Adolf, G. R., Maurer-Fogy, I., Kalsner, I. & Cantell, K. (1990) J. Biol. Chem. 265, 9290-9295], the insect cell produced protein which was demonstrated by lectin blot, mass spectroscopy and HPLC analysis to contain only the core oligosaccharide. Two different structures, (Man)2(GlcNAc)2[Fuc] and (Man)3(GlcNAc)2[Fuc] were identified. The fucosylation was identified to be (alpha 1-6)-linked to the core saccharide. Sialic acid residues were clearly absent. IFN-omega 1 expressed in S. frugiperda cells was shown to be partially truncated at the C-terminus by nine residues; its antiviral activity when glycosylated was significantly lower than the activity of IFN-omega 1 produced by Sendai-virus-stimulated leukocytes. Circular dichroism and fluorescence spectroscopy did not reveal any structural differences between glycosylated and nonglycosylated IFN-omega 1. This implies the importance of a complex-type glycosylation for the maximal biological activity of human IFN-omega 1. PMID- 8223649 TI - Expression of human interferon-alpha 2 in Sf9 cells. Characterization of O-linked glycosylation and protein heterogeneities. AB - Human interferon alpha 2 (IFN-alpha 2) was expressed in Spodoptera frugiperda Sf9 insect cells using the baculovirus expression system. The protein purified by immunoaffinity chromatography exhibited biological activity identical to that of leukocyte-derived 'natural' IFN-alpha 2. However, the protein was found to be heterogeneously glycosylated, partially truncated by proteolysis and partially lacking a disulfide bridge. The major product was shown to be O-glycosylated at the same position as natural human IFN-alpha 2. Enzymatic cleavage, reverse-phase HPLC peptide mapping and plasma-desorption mass spectroscopy analysis revealed the presence of two types of O-linked carbohydrates. The major O-linked carbohydrate was found to be the disaccharide galactosyl(beta 1-3)-N acetylgalactosamine, the minor component the monosaccharide N acetylgalactosamine. No evidence for sialylation was found. The non-glycosylated species representing about 40% of the total material were shown to partially lack the C-terminal three amino acids. In addition an unglycosylated, reduction sensitive dimer was observed. This was formed due to the lack of the N-terminal cysteine normally forming an intramolecular disulfide bridge. Furthermore, a minor species was identified which contains Cys1 and Cys98 in a modified form, thereby hindering the formation of a disulfide bridge between these two residues. PMID- 8223650 TI - Purification of the large mobilization protein of plasmid RSF1010 and characterization of its site-specific DNA-cleaving/DNA-joining activity. AB - A site-specific and strand-specific nick, introduced into the RSF1010 plasmid origin of transfer (oriT), initiates unidirectional DNA transfer during bacterial conjugation. We have previously reproduced this nicking at the duplex oriT in vitro using purified preparations of the three known RSF1010-mobilization proteins: MobA (78-kDa form of RSF1010 primase), MobB and MobC [Scherzinger, E., Lurz, R., Otto, S. & Dobrinski, B. (1992) Nucleic Acids Res. 20, 41-48]. In this study we report the purification of MobA to apparent homogeneity and demonstrate that this 78-kDa protein by itself is capable of creating the oriT-specific nick if the DNA is present in the single-stranded form. By studying the cleavage of sets of oligodeoxyribonucleotides varying successively by single nucleotides at the 5' or 3' end, the minimal substrate for cleavage has been defined. The results identify the MobA recognition sequence within the 11-residue oligonucleotide AAGTGCGC-CCT which is cleaved at the 3' side of the G at position 7. During the cleavage reaction, MobA becomes covalently linked to the 5' phosphate end of each broken DNA molecule and retains its activity for the rejoining reaction. It can transfer the attached DNA to an incoming acceptor strand provided that the DNA molecule contains at its 3' end at least the seven nucleotides upstream of the nick site. The covalent MobA-DNA linkage has been determined by two-dimensional thin-layer electrophoresis to be a tyrosyl phosphate. Extensive digestion of the 32P-labeled MobA-oligonucleotide complex with lysine carboxypeptidase yielded a single DNA-bound peptide which was purified and sequenced. The resulting peptide sequence consists of amino acid residues at positions 22-30 in the MobA sequence and identifies Tyr24 as the residue linked to DNA in the covalent complex. PMID- 8223651 TI - Site-directed mutagenesis of beta-lactamase TEM-1. Investigating the potential role of specific residues on the activity of Pseudomonas-specific enzymes. AB - From sequence alignments, two groups can be defined for the carbenicillin hydrolysing beta-lactamases (CARB enzymes). One group includes the Pseudomonas specific enzymes PSE-1, PSE-4, CARB-3, CARB-4 and also the Proteus mirabilis GN79, for which the well-conserved residue Lys 234 in all class-A beta-lactamases is changed to an arginine residue. The second group includes the enzymes PSE-3 and AER-1 which have an arginine or a lysine residue at position 165. All these enzymes also have leucine at position 68, threonine at position 104 and glycine at position 240. We engineered these mutations into the TEM-1 beta-lactamase to study their potential role in defining the substrate profile of the CARB enzymes. The mutations K234R and E240G in TEM-1 noticeably increased the hydrolysis of carboxypenicillins relative to other penicillins by approximately sixfold and twofold, respectively. The variant E240G also demonstrated an improved rate of second-generation cephalosporin and cefotaxime hydrolysis. In contrast, the substitution of Trp165 by arginine does not extend the substrate profile to alpha carboxypenicillins nor does it noticeably modify the kinetic behavior of the enzyme. The mutations M68L and E104T do not have a large effect on the hydrolysis rate but the mutation E104T enhances the affinity of the enzyme for third generation cephalosporins. As the mutation K234R resulted in a severe decrease in the affinity for carboxypenicillins, the double mutant E240G/K234R was constructed in an attempt to enhance the CARB character of the enzyme. Contrary to what could be expected, the additional mutation E240G for the TEM-1 K234R enzyme increases neither the catalytic constant for the carboxypenicillins nor the affinity towards these substrates. Consequently, this study strongly suggests that the three-dimensional structures of the active site of the TEM-1 enzyme and PSE-3, PSE-4 or other related enzymes are significantly different. This probably explains the discrepancy of the substrate profile between the CARB enzymes and the TEM-1 protein variants. PMID- 8223652 TI - Stereochemistry, specificity and kinetics of the hydrolysis of reduced cellodextrins by nine cellulases. AB - The catalytic activity of nine enzymes (endoglucanases I-III, V, VI and cellobiohydrolases I and II from Humicola insolens; endoglucanases A and C from Bacillus lautus), representative of cellulase families A-C, H, J and K, has been investigated using a series of reduced cellooligosaccharides (cellotriitol to cellohexaitol) as substrates. For each enzyme, the specificity of cleavage was determined by analytical HPLC while the kinetic constants were obtained from a kinetic assay involving a cellobiose dehydrogenase purified from H. insolens as a coupled enzyme using 2,6-dichloroindophenol as the electron acceptor. These data were used to estimate the number of subsites in the enzymes. The stereochemical course of hydrolysis by seven enzymes, representing the six different families, was assessed using 1H-NMR. The enzymes belonging to families which had already been investigated (A-C), showed results in agreement with previous studies. The three other families (H, J and K), for which no mechanistic data was previously available, gave results which indicated that enzymes in group H had retaining type activity and enzymes in groups J and K had inverting-type activity. The retaining endoglucanases I and III displayed a high glycosyl-transferase activity under the conditions used during the NMR experiments resulting in precipitates of higher oligomers. PMID- 8223653 TI - The second nucleophile molecule binds to the acyl-enzyme-nucleophile complex in alpha-chymotrypsin catalysis. Kinetic evidence for the interaction. AB - alpha-Chymotrypsin-catalyzed acyl transfer was studied using three acyl-group donors (Mal-L-Ala-L-Ala-L-PheOMe, Bz-L-TyrOEt and Ac-L-TrpOEt; Mal, maleyl; Bz, benzoyl; OMe, methyl ester; OEt, ethyl ester) and a series of amino-acid amides. Most of the reactions studied can be described by the simplest kinetic model without the nucleophile binding to the acyl-enzyme. The alpha-chymotrypsin catalyzed transfer of the Mal-L-Ala-L-Ala-L-Phe group to the amides of L-Phe and L-Tyr showed a linear dependence of the partition constant, p, on the nucleophile concentration which can be interpreted by the hydrolysis of the acyl-enzyme nucleophile complex. The alpha-chymotrypsin-catalyzed transfer of the Bz-L-Tyr and Ac-L-Trp groups to several amino-acid amides showed unusual behavior which can be interpreted by the kinetic model involving formation of a complex of the acyl-enzyme with two nucleophile molecules. These observations can explain the conflicting conclusions concerning the kinetics of alpha-chymotrypsin-catalyzed acyl transfer evident in previous studies. PMID- 8223654 TI - Proteolytic cleavage of synthetic fragments of vesicle-associated membrane protein, isoform-2 by botulinum type B neurotoxin. AB - Recent data suggest that botulinum type-B neurotoxin is a protease which acts on vesicle-associated membrane protein, isoform 2 (VAMP-2). In this report, botulinum type-B neurotoxin is shown to cleave a synthetic fragment (HV62) of VAMP-2, corresponding to the bulk of the hydrophilic domain (amino acids 33-94). The neurotoxin acts at a single site between Gln76 and Phe77. Little or no proteolytic activity by botulinum type-B neurotoxin was observed with peptides containing 7, 10 or 20 amino acids spanning the site of cleavage. The proteolytic action of neurotoxin was strongly inhibited by EDTA and o-phenanthroline whereas captopril and phosphoramidon were ineffective. A series of model peptide substrates were synthesised in order to define the smallest VAMP-2 fragment to be cleaved by botulinum type-B neurotoxin. Data obtained from these substrates suggest that the neurotoxin belongs to a novel class of zinc-endoprotease; more than 12 amino acid residues are required on both the NH2- and COOH-terminal side of the cleavage site for optimal proteolytic activity. The results demonstrate that no other components of cellular vesicles are required for the specific action of the neurotoxin on VAMP-2. The data further show that the highly specific action of the neurotoxin is not dictated solely by the properties of the amino acid residues at the cleavage site but is also dependent on amino acid sequences distal to its site of action. PMID- 8223655 TI - Dihydrorhodamine 123: a fluorescent probe for superoxide generation? AB - Imaging techniques, such as confocal microscopy and fluorescent activated cells scan are facilitating the study of responses at the single-cell level. Superoxide is reported to oxidise the non-fluorescent dihydrorhodamine 123 (DHR) to rhodamine 123. The generation of rhodamine 123 by human neutrophils, stimulated by the phorbol ester phorbol 12-myristate 13-acetate was inhibited slowly by diphenylene iodonium and rapidly by azide, but not by superoxide dismutase. In the absence of enzymes H2O2 (but not O2-.) oxidised DHR slowly but the rate was greatly enhanced by peroxidases. The rhodamine 123 generated by phorbol-ester stimulated neutrophils was observed to be located within the cell despite the fact that neutrophils failed to accumulate external rhodamine 123. This stimulated rise in cellular fluorescence was eliminated by excess extracellular catalase. It appears that H2O2, released on the outside, crosses the plasma membrane where oxidation of DHR is catalysed by cellular peroxidases. Since in a mixed population DHR failed to distinguish between O2-.-producing and non producing HL60 cells it is not a suitable probe for single-cell observations. We conclude that DHR oxidation reports only the presence of H2O2 and intracellular peroxidases, and not the generation of O2-. by any one cell. Only peroxidase containing cells fluoresce. PMID- 8223656 TI - Voltammetric studies of the catalytic electron-transfer process between the Desulfovibrio gigas hydrogenase and small proteins isolated from the same genus. AB - The kinetics of electron transfer between the Desulfovibrio gigas hydrogenase and several electron-transfer proteins from Desulfovibrio species were investigated by cyclic voltammetry, square-wave voltammetry and chronoamperometry. The cytochrome c3 from Desulfovibrio vulgaris (Hildenborough), Desulfovibrio desulfuricans (Norway 4), Desulfovibrio desulfuricans (American Type Culture Collection 27774) and D. gigas (NCIB 9332) were used as redox carriers. They differ in their redox potentials and isoelectric point. Depending on the pH, all the reduced forms of these cytochromes were effective in electron exchange with hydrogenase. Other small electron-transfer proteins such as ferredoxin I, ferredoxin II and rubredoxin from D. gigas were tentatively used as redox carriers. Only ferredoxin II was effective in mediating electron exchange between hydrogenase and the working electrode. The second-order rate constants k for the reaction between reduced proteins and hydrogenase were calculated based on the theory of the simplest electrocatalytic mechanism [Moreno, C., Costa, C., Moura, I., Le Gall, J., Liu, M. Y., Payne, W. J., van Dijk, C. & Moura, J. J. G. (1993) Eur. J. Biochem. 212, 79-86] and the results obtained by cyclic voltammetry were compared with those obtained by chronoamperometry. Values for k of 10(5)-10(6) M 1 s-1 (cytochrome c3 as electron carrier) and 10(4) M-1 s-1 (ferredoxin II as the electron carrier) were determined. The rate-constant values are discussed in terms of the existence of an electrostatic interaction between the electrode surface and the redox carrier and between the redox carrier and a positively charged part of the enzyme. PMID- 8223657 TI - Catalytic mechanism of 3-deoxy-D-manno-2-octulosonate-8-phosphate synthase. The use of synthetic analogues to probe the structure of the putative reaction intermediate. AB - The proposed mechanistic pathway for the reaction catalyzed by 3-deoxy-D-manno-2 octulosonate-8-phosphate (Kdo8P) synthase was examined in terms of the structure of the putative bisphosphate intermediate. Two 2-deoxy analogues of the product Kdo8P, having been structurally prohibited from undergoing the ring-opening and possessing the stereochemistry of either the alpha-pyranase (compound 1) or the beta-pyranose form (compound 2) of the product, were synthesized and probed as inhibitors for the synthase. It was found that both analogues bind to the enzyme and are competitive inhibitors with respect to phosphoenolpyruvate binding, having Ki values of 470 microM and 303 microM, respectively. Comparison of this data to the Ki value of the tautomeric mixture of the product Kdo8P (Ki = 590 microM) suggests that both the alpha- and the beta-pyranose anomers (65.8% and 3.1%, respectively at neutral pH) bind to the enzyme with a slight (1.13 kJ/mol) preference for the beta-anomer, and that the C2 hydroxyl does not contribute to the binding. This uncertain stereochemical preference exhibited by the enzyme for the stereoisomers at the anomeric carbon suggests that the carboxylate binding site of the product is indistinct, while the hydroxyl and carboxylate binding sites may be interchangeable. More importantly, however, the isosteric phosphonate analogue 2,6-anhydro-3-deoxy-2 beta-phosphonylmethyl-8-phosphate-D glycero-D-talo-octonate (3), which mimics the topological and electrostatic properties of the proposed cyclic intermediate, was found to be the most potent inhibitor of the enzyme with a Ki value of 5 microM. Two hitherto unrecognized aspects of the mechanism of the synthase were identified. First, the results showing that the cyclic analogues 1, 2 and 3 are inhibitors of the enzyme whereas the previously reported acyclic analogue, which contains no carbonyl group at C2 and may thus resemble the open-chain form of Kdo8P, is not an inhibitor, suggest that the pyranose form and not the open-chain acyclic form of the putative bisphosphate intermediate is handled by the enzyme. Second, since the overall stereochemical course of the transformation mediated by the synthase has been shown to involve si face addition of phosphoenolpyruvate to the re face of the carbonyl of arabinose 5-phosphate, the present observation involving analogue 3 suggest that the bisphosphate intermediate formed during the initial steps of synthesis may have the pyranose structure with the anomeric phosphate located in the beta-configuration. PMID- 8223658 TI - Western blot analysis of antibody responses of young infants to pertussis infection. AB - Western blot and agglutination techniques were used to analyze the antibody responses to Bordetella pertussis in 27 infants less than six month of age with presumed pertussis infection. The antibody response to the Bordetella pertussis adhesions filamentous hemagglutinin, pertactin and agglutinogens, and to the Bordetella pertussis toxins pertussis toxin and adenylate cyclase-hemolysin were compared. Infection induced intense antibody responses to filamentous hemagglutinin, pertussis toxin and adenylate cyclase-hemolysin. Antibodies to agglutinogens were never detected, and antibodies to pertactin were rarely detected in infected infants' sera. Therefore, determination of anti agglutinogens levels only is not suitable for the serological diagnosis of pertussis in young infants. Use of purified filamentous hemagglutinin, pertussis toxin and adenylate cyclase-hemolysin in Western blot analysis may improve the serodiagnosis of Bordetella infections. However, care must be exercised in distinguishing between the antibody response in young infants and maternally derived antibodies. PMID- 8223659 TI - Imipenem resistance in Enterobacter. AB - Blood cultures obtained on two separate occasions from a 37-year-old male who received multiple antibiotics (including imipenem) for treatment of repeated episodes of intraabdominal abscesses and bacteremia yielded two isolates of Enterobacter with reduced susceptibility to imipenem, extended-spectrum cephalosporins, penicillins and aztreonam. Both isolates were unstable, giving rise to different colony types, each of which produced a single, non-inducible Bush group 1 beta-lactamase (pI = 9.6) that hydrolyzed imipenem. Outer membrane proteins were analyzed but no differences were detected between strains with different levels of imipenem resistance. Three-dimensional tests performed in conjunction with disk diffusion susceptibility tests provided a rapid and convenient means of detecting the production of imipenem-hydrolyzing enzymes by the Enterobacter strains. These isolates provided additional evidence that overproduction of the group 1 cephalosporinase of Enterobacter can contribute to resistance to imipenem. PMID- 8223660 TI - Oral itraconazole plus nasal amphotericin B for prophylaxis of invasive aspergillosis in patients with hematological malignancies. AB - The use of oral itraconazole (200 mg daily) plus nasal amphotericin B (10 mg daily) for prophylaxis of invasive aspergillosis was evaluated in 164 patients with hematological malignancies at risk due to presence of neutropenia and/or steroid therapy. This prophylactic regimen was evaluated for a period of two years. Two hundred and ninety patients with similar characteristics who were observed over the three-year period prior to the introduction of prophylaxis served as historical control group. Environmental surveillance during the study period showed constant contamination of the air with Aspergillus. Prophylaxis significantly reduced the incidence of proven invasive aspergillosis from 12/290 to 0/164 (p = 0.004), and reduced the mortality rate from 8/290 to 0/164. The incidence of proven plus probable aspergillosis amounted to 34/290 in the control group and 8/164 in the study group (p = 0.01); the mortality rates were 11/290 (3.7%) and 2/164 (1.2%) respectively. All nasal cultures in the study group were negative for Aspergillus. The prophylactic regimen was well tolerated. Larger studies assessing each agent alone and in combination are necessary to confirm these observations. PMID- 8223661 TI - Computerised calculation of the true costs of antibiotic therapy. AB - A system for computerised calculation of the drug costs and hidden costs of antibiotic therapy is described. A previously developed method for costing antibiotic therapy was used to quantify hidden costs arising from intravenous administration, labour, serum antibiotic assays, monitoring of haematological and biochemical indices, and disposal of sharp instruments. These calculations were installed into a software system using spreadsheets (Microsoft Excel/Windows). Cost calculations were simplified as a result of automated calculation with centralised cost updating, a facility for new antibiotics, a facility for atypical regimens, availability to a number of users, password protection and a facility for multiple courses of intravenous antibiotic therapy. PMID- 8223663 TI - Use of nonradioactive DNA probes to identify a Campylobacter jejuni strain causing abortion. AB - A case of human abortion due to a Campylobacter infection is reported. Cultures revealed two morphologically different isolates with large and small colonies respectively. Using conventional methods of identification, the large colonies were identified as Campylobacter jejuni and the small colonies as Campylobacter coli. Dot blot hybridization and determination of rDNA restriction fragment patterns revealed that both colony types were the same strain of Campylobacter jejuni. This observation illustrates the need to use methods other than phenotypic methods when identifying strains of Campylobacter. PMID- 8223662 TI - Endocarditis caused by Neisseria elongata subspecies nitroreducens: case report and literature review. AB - Neisseria elongata subsp. nitroreducens, an usual commensal of the nasopharynx, can cause serious infections, usually endocarditis. A case of Neisseria elongata endocarditis complicating mitral valve prolapse in a patient allergic to penicillin is reported. Cure was obtained with a combination of ceftriaxone and gentamicin, there being no requirement for surgery during the acute phase of the disease. PMID- 8223664 TI - Evaluation of a colour test for rapid detection of Salmonella. AB - The performance of a colour test (Wellcolex) for rapid detection of the most frequently isolated O serogroups of salmonella (A, B, C, D, E/G) and the Vi antigen was evaluated using colonies grown on enteric differential agar media and in selenite broth. The test had excellent sensitivity (100%) and specificity (100%) with colonies taken directly from primary culture plates. Initial tests in selenite broth showed limited sensitivity (62.1%) but by using high quality media and modifying the inoculation procedure, sensitivity was greatly improved (99%). PMID- 8223665 TI - Evaluation of a commercial latex agglutination test for rapid detection of Salmonella in fecal samples. AB - A latex agglutination test for the detection of salmonella in feces was evaluated in comparison to direct culture and enriched culture using both artificially inoculated samples and clinical samples. In the samples inoculated artificially with different concentrations of salmonella (10(1) to 10(5) per gram) the enriched culture performed better only at the 10(2) level in 0.4 g samples, whereas the latex test performed as well as the enriched culture at all levels in 4 g samples. In the tests using clinical samples, there was no significant difference between results of the latex test performed in 2283 samples and the enriched culture performed in 2072 samples. The sensitivity, specificity and negative and positive predictive values of the latex test were 88.2%, 98%, 97.5% and 63% respectively. The test provided results rapidly but yielded a number of false positive results. PMID- 8223666 TI - Comparison of a commercial enzyme immunoassay and an immunoblot technique for detection of immunoglobulin A antibodies to Toxoplasma gondii. AB - A commercial enzyme immunoassay (Platelia-Toxo IgA) and an immunoblot technique were compared with regard to their ability to detect IgA antibodies to the major surface protein P30(SAG1) of Toxoplasma gondii in 105 serum samples from patients with suspected or proven acquired toxoplasmosis. Comparison of the IgA-EIA with the immunoblot technique showed a concordance of 81.0%, with a sensitivity of 92.6% and a specificity of 78.4%. Due to its high sensitivity the IgA-EIA might detect IgA antibodies against Toxoplasma gondii at an early stage of infection, although excessive sensitivity could lead to detection of IgA antibodies for an extended period of time following the onset of infection. PMID- 8223667 TI - In vitro activity of the new quinolone BAY y 3118 against anaerobic bacteria. AB - The in vitro activity of BAY y 3118 against anaerobic cocci, Propionibacterium acnes, Clostridium perfringens, Clostridium difficile, Bacteroides fragilis, other Bacteroides spp. and fusobacteria was determined by an agar dilution method. This activity was compared with that of ciprofloxacin, ofloxacin, piperacillin, cefoxitin, imipenem, clindamycin and metronidazole. BAY y 3118, imipenem, clindamycin and metronidazole were the most active agents tested. The in vitro activity of BAY y 3118 against anaerobic bacteria was superior to that of ciprofloxacin and ofloxacin. PMID- 8223668 TI - Vertebral osteomyelitis caused by Haemophilus aphrophilus. PMID- 8223669 TI - Meningitis due to Gemella haemolysans. PMID- 8223670 TI - Increased resistance of enteropathogens to fluoroquinolones in Barcelona, Spain. PMID- 8223671 TI - Synergistic activity of tobramycin-netilmicin against Enterococcus faecalis producing 6'-aminoglycoside-acetyltransferase, 2''-aminoglycoside phosphotransferase enzyme. PMID- 8223672 TI - Isolation of vancomycin-resistant lactobacilli from three neutropenic patients with pneumonia. PMID- 8223673 TI - Serological markers for diagnosis of acute delta hepatitis infection. PMID- 8223674 TI - Evaluation of urogenital Chlamydia trachomatic infections by cell culture and the polymerase chain reaction using a closed system. PMID- 8223675 TI - Pneumatic phlebography: a possible new technique for the assessment of recurrent varicose veins. AB - Initial experience with a new technique, using an inflatable compression device during ascending phlebography, is described. This method is compared with standard varicography in a small series of patients with recurrent varicose veins. In this pilot study, 13 patients had 17 limbs examined by the new method and 14 limbs by varicography. Below knee perforator incompetence was demonstrated directly in all limbs by 'pneumatic phlebography'. Short-saphenous vein and mid thigh perforator incompetence were also more frequently demonstrated. The two tests are comparable at showing recurrent or persistent connection at the groin (13 limbs, overall). With modifications of the pressure device, pneumatic phlebography may prove to be an important method of assessing recurrent varicose veins. PMID- 8223676 TI - 99mTc-DTPA activity in the azygos vein before and after sclerotherapy in patients with cirrhosis. AB - We determined the radioactivity of 99mTc-diethylene-triamine pentaacetic acid (99mTc-DTPA) from the azygos vein in 20 patients with cirrhosis to evaluate the hemodynamic relationship between esophageal varices and azygos venous blood flow before and after sclerotherapy. 99mTc-DTPA was injected into the splenic artery before and after sclerotherapy through a catheter in the right femoral artery. The patients were classified into three types. Type I consisted of nine patients who showed maximum levels of 99mTc-DTPA which were significantly lower (P < 0.01) than those determined before sclerotherapy. The Type II was composed of six patients who had far lower radioactivity levels than those of Type I, but also showed significantly lower maximum activity after sclerotherapy than before (P < 0.01). The five patients in Type III had significantly higher radioactivity after sclerotherapy than before (P < 0.05). Rupture of the esophageal varices occurred in one of the Type I (11.1%), none of the Type II, and four of Type III (80%) cases, among all of the patients followed for 40 months after sclerotherapy. The radioactivity in the azygos vein appeared later in Type I (P < 0.05) and Type II after sclerotherapy than before sclerotherapy, but in Type III there was no difference between pre- and post-sclerotherapy values. Type III patients in whom portography was performed showed short gastric and paraesophageal veins, whereas Type I and Type II patients did not. These data suggest that radioactivity in the azygos vein before and after sclerotherapy reflect the grade of shunting and estimate the recurrence of bleeding in patients with cirrhosis associated with esophageal varices. PMID- 8223677 TI - Technical difficulty of femoral artery puncture in late pregnancy. PMID- 8223678 TI - Primary retroperitoneal serous cystadenocarcinoma of 'ovarian-type': US and CT findings. PMID- 8223679 TI - Fat suppression techniques in MR imaging of mature ovarian teratomas: comparison with CT. AB - OBJECTIVE: The aim of this study is to analyze the ability of MR fat suppression techniques to characterize fat components of ovarian mature cystic teratomas (MCT) shown by CT. SUBJECTS AND METHODS: MR images of eight MCTs of the ovary were obtained using standard sequences followed by a SPIR (spectral presaturation with inversion recovery) sequence in six cases and by Dixon sequences in two cases. In all cases correlation with CT and pathologic findings was achieved. RESULTS: MR fat suppression sequences showed to be as accurate as CT in detecting fat inside the cystic part of the teratomas (8/8). CONCLUSION: MR fat suppression sequences should be performed when presence of fat is suspected on images of ovarian tumors produced by standard MR sequences. PMID- 8223680 TI - A prospective study of the value of mammographic patterns as indicators of breast cancer risk in a screening experience. AB - The association between mammographic parenchymal patterns and the risk of subsequent breast cancer was studied in a cohort of 17,911 women aged 40-70 years, enrolled in a mammographic screening program. The cohort was prospectively followed-up for 5 years after blind attribution of mammographic pattern. Incident cancers, either screen- or interval-detected, were monitored through the screening program and by the local cancer registry. Multivariate analysis adjusting for the confounding effect of age, demonstrated an independent and significant association between mammographic patterns and breast cancer risk. The relative risk of subsequent breast cancer, assumed to be 1 for N1 pattern, was 2.39 (1.0-5.7) for P1, 4.02 (1.7-9.3) for P2, and 5.01 (1.9-13.3) for DY category, respectively. The relative risk of P2-DY with respect to N1-P1 categories was 2.09 (1.4-3.1). A diagnostic delay, possibly due to the 'masking effect' of a radiologically dense breast, was evident for P2-DY with respect to N1-P1 cancers (proportion of Stage I cancers = 42% vs. 69%). These results suggest the adoption of a shorter rescreening interval in P2-DY subjects. PMID- 8223681 TI - Duplex-Doppler ultrasonography in the assessment of portal hypertension. Utility of the measurement of maximum portal flow velocity. AB - To assess the utility of Duplex-Doppler ultrasonography (DDUS) in the evaluation of portal haemodynamics we studied 52 patients with compensated liver cirrhosis (mean age 53.9 +/- 9.2 years, males 32, females 20) diagnosed by laparoscopy and biopsy. All patients underwent laparoscopy and oesophageal-gastro-duodenoscopy (OGDS): we postulated that haemodynamically significant portal hypertension (PH) was present if varices were detected at OGDS and/or if collateral veins were revealed at laparoscopy. DDUS was performed with a strictly standardized method and maximum portal flow velocity (PFV) was measured in all patients. Max-PFV ranged between 3.5 and 33.4 cm/s. Overall, 36 patients (69%) had a max-PFV lower than 20.3 cm/s (normal max-PFV range in our laboratory is 20.3-33.3 cm/s), while 16 patients (31%) had normal max-PFV values. Five patients (9.6%) had no signs of PH at laparoscopy and/or OGDS and all five had normal max-PFV values. The other 47 patients (90.4%) had collateral circuli at laparoscopy and 29/47 (61.7%) exhibited also varices at OGDS: max-PFV was lower than 20.3 cm/s in 36/47 patients (76.6%). The measurement of max-PFV demonstrated a 76.6% sensitivity and a 100% specificity in detecting PH, with 100% positive predictive value and 31% negative predictive value. Three patients with PH and apparently normal max-PFV values exhibited a recanalization of the umbilical vein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223682 TI - Polyarthropathy--a delayed reaction to low osmolality angiographic contrast medium in patients with end stage renal disease. AB - Since the introduction of low osmolality non-ionic media, acute reactions to radiographic contrast are uncommon and delayed reactions are especially rare, consisting mostly of mild flu-like symptoms. We report two patients suffering from end-stage renal failure and treated by continuous ambulatory peritoneal dialysis (CAPD) who developed a severe constitutional illness including acute polyarthropathy 6 and 16 h after injection of the low osmolality non-ionic contrast medium, iopamidol. Although the clinical presentation of the reactions was similar to a systemic lupus syndrome there was no immunological evidence to support this as an aetiological mechanism. Since CAPD is a relatively inefficient method of clearing contrast media prolonged high circulating levels of iopamidol may have been a contributory factor to these unusually severe delayed contrast reactions. PMID- 8223683 TI - Ultrasound and hip joint effusion. PMID- 8223684 TI - Ultrasound and hip joint effusion. PMID- 8223685 TI - Pediatric bronchography performed through the flexible bronchoscope. PMID- 8223686 TI - The American Thoracic Society lymph node map: a CT demonstration. AB - It is now a decade since the American Thoracic Society (ATS) lymph node map was first described. Recently it has been upgraded to include supraclavicular and superior diaphragmatic lymph nodes. By standardising terminology the map aids in the provision of a more valid data base to evaluate diagnostic techniques and therapeutic protocols. Despite these potential benefits the map has failed to gain widespread use amongst European radiologists. This essay illustrates the relationship of the nodal stations to normal anatomy utilizing calcified nodes on CT scans. The ease with which the map can be applied to cross-sectional anatomy is emphasized. It is hoped that the images will act as a teaching aid to promote greater acceptance and use of the ATS map. The relationship of the nodal stations to the new American Joint Classification of disease extent is also outlined. PMID- 8223687 TI - Cystic lesions of the brain. A classification based on pathogenesis, with consideration of histological and radiological features. AB - A classification of the existing multitude of cystic lesions of the brain is proposed, which allows an understanding of their genesis and consequent therapeutic implications, as well as their diagnostic characteristics. Essentially, cerebral cystic lesions may be classified into the following categories: Cysts containing CSF-like fluid, which include ex vacuo type cysts, such as leptomeningeal cysts, and cysts following surgical resection; cysts with fluid secreting walls and CSF-like content, such as arachnoid cysts; cysts associated with dysgenesis, for example Dandy-Walker cysts. The ex vacuo cysts increase craniospinal compliance, whereas the other cysts with CSF-like content do not; they are not per se expansive, however, although their occasional location along CSF pathways may cause obstruction and hydrocephalus. Another category includes cysts with a lining of non-neural epithelium like colloid cysts, epidermoid cysts, or craniopharyngiomas. They may increase in size and cause symptoms by compression, although not at the rate of tumour-associated cysts. The cysts associated with gliomas and other tumours have a pathogenesis bearing upon blood-brain barrier impairment and formation of vasogenic oedema. Finally, one may distinguish a category of cysts with infectious origin, such as brain abscesses and hydatid cysts. The cysts with CSF-like contents may be recognised by their magnetic resonance characteristics resembling those of CSF, whereas cysts containing proteinaceous fluid are associated with blood-brain barrier impairment and consequent contrast enhancement. The cysts with a lining of non-neural epithelium exhibit diverse properties of attenuation on computed tomography (CT) and magnetic resonance imaging (MRI), depending on the nature of their cyst contents. PMID- 8223688 TI - Orbitocranial penetration of a pencil: extraction under CT control. PMID- 8223689 TI - Findings of focal hepatic lymphoma on Gd-DOTA enhanced dynamic FLASH MR imaging. PMID- 8223690 TI - MR imaging of vertebral metastases: evaluation of fat saturation imaging. AB - OBJECTIVE: To compare conventional T1-weighted imaging (SE-T1) and chemical shift fat-saturation T1-weighted imaging (FS-T1) in the diagnosis of bone metastases. SUBJECTS: Twenty-two patients (101 vertebrae) with non-neoplastic lesions (Group 1) and 21 patients (72 vertebrae) with spinal metastases (Group 2) were evaluated with both images. RESULTS: The signal intensity (SI) distribution of Groups 1 and 2 on SE-T1 showed various patterns, and SI measurement was not significantly different between the two groups. On FS-T1, all non-neoplastic lesions had a low intensity homogeneous appearance; however, the metastases had mixed to high SI. The SI measurement data of Group 2 was significantly (P < 0.0001) higher than that of Group 1. CONCLUSION: FS-T1 was useful in evaluating vertebral metastases. PMID- 8223691 TI - Display of MR angiograms: maximum intensity projection versus three-dimensional rendering. AB - OBJECTIVE: The purpose of this study is to clarify the most useful technique of MR imaging in demonstrating the collateral venous pathways due to portal hypertension. SUBJECTS: Twenty-eight cirrhotic patients with collateral venous pathways due to portal hypertension and three patients without them were studied. We evaluated the detectability of collateral vessels of flow-enhanced transverse MR images by time of flight effect, projection MR angiography using the maximum intensity projection technique and three-dimensional (3D) computer graphic images, by using the arterial portography as the standard. RESULTS: Flow-enhanced transverse MR images showed 98% of the collateral vessels detected by angiography, however, the connection of each vessel was obscure. Fully 81% of the collateral vessels were seen by projection MR angiography, and 98% of them by the 3D computer graphic images. CONCLUSION: The detectability of collateral vessels of 3D computer graphic imaging was better than that of the projection MR angiography and was as accurate as that of transverse MR imaging. 3D Computer graphic images provided complex anatomical information in a more easily understood fashion than either transverse MR images or projection MR angiography. PMID- 8223693 TI - Isolation and characterization of the Golgi apparatus of a flagellate scaly green alga. AB - Highly purified Golgi membranes were isolated from the scaly green flagellate Scherffelia dubia using osmotic shock for controlled cell rupture, differential centrifugations and a discontinuous sucrose density gradient centrifugation. Three Golgi membrane fractions (based on the distribution of IDPase activity in the gradient) at densities 1.14 g/ml, 1.17 g/ml and 1.20 g/ml were obtained. The specific IDPase activity in these fractions was enriched about 78-fold compared to the crude cell homogenate. The Golgi membrane fractions were further characterized by electron microscopy, SDS-PAGE and lectin blotting. The low density fraction (1.14 g/ml) contained two distinct vesicle populations and scale precursors associated with the outer surface of the larger-size vesicles. The medium density fraction (1.17 g/ml) contained in addition to the larger vesicles, multilamellate vesicles and semicircular cisternae. Finally, in the high density fraction (1.20 g/ml) in addition to small and large vesicles, a tubular membrane reticulum was observed. The three Golgi membrane fractions revealed the same complex overall polypeptide composition when analyzed by SDS-PAGE, but gradual quantitative differences in the polypeptide profile between fractions were observed. The lectins GNA, DSA, and AAA bound to several glycoproteins in all Golgi membrane fractions. Deglycosylation with N-glycosidase F showed that all carbohydrate structures recognized by GNA and DSA, and one recognized by AAA were of the N-glycosidic type indicating the presence of both "high mannose" and "processed" N-glycans in the Golgi apparatus of S. dubia. PMID- 8223692 TI - ERGIC-53, a membrane protein of the ER-Golgi intermediate compartment, carries an ER retention motif. AB - Overlapping cDNAs encoding the entire human ERGIC-53, a 53 kDa membrane protein of the ER-Golgi intermediate compartment, have been isolated and their nucleotide sequence determined. The isolated cDNA is about 2.7 kb in length. The deduced polypeptide chain for ERGIC-53 consists of 510 amino acids (M(r) 54217) including an N-terminal signal sequence of 30 amino acids, a single putative transmembrane segment of 18 amino acids, and a short cytoplasmic domain of 12 amino acids. Surprisingly, the cytoplasmic segment contains two lysines positioned three and four residues from the C-terminus. Such a double lysine motif is known to function as a retention signal for a group of membrane proteins associated with the ER. Expression of a full-length cDNA of ERGIC-53 in Vero cells revealed intracellular localization similar but not always identical to the endogenously expressed ERGIC-53. The presence of an ER retention motif in a protein of the ER Golgi intermediate compartment has important implications for the retention mechanism mediated by this signal. PMID- 8223694 TI - Age-related changes in expression of neural cell adhesion molecule (NCAM) in heart: a comparative study of newborn, adult and aged rats. AB - The neural cell adhesion molecule (NCAM) has been implicated in cellular interactions involved in cardiac morphogenesis and innervation. In this study, expression of NCAM mRNA and protein was characterized in rat heart during postnatal development and aging (postnatal days 1, 10, 40, 270, and 730). Alternative splicing of NCAM mRNA was analyzed by Northern blotting using DNA oligonucleotide probes designed for demonstration of certain exons or exon combinations. Total NCAM mRNA was downregulated during postnatal development followed by upregulation in the aging heart. Three major NCAM mRNA classes of 6.7, 5.2 and 2.9 kb were expressed in newborn heart in approximately equal proportions. At all other ages, the mRNAs of 5.2 and 2.9 kb were more predominant than the 6.7 kb mRNA. During postnatal development and aging, expression of exon VASE was selectively downregulated in the 6.7 kb NCAM mRNA class, whereas it was clearly detectable in NCAM mRNA classes of 5.2 and 2.9 kb. Insertion of exons a and AAG between exons 12 and 13 was more pronounced in the 5.2 and 2.9 kb NCAM mRNAs than in the 6.7 kb mRNA at all ages. Insertions at the 12/13 junctions decreased in the 6.7 kb mRNA as compared to the 5.2 and 2.9 kb mRNAs during postnatal development. Quantification of NCAM protein by enzyme-linked immunosorbent assay showed that NCAM protein amount decreased from a level of 0.93 microgram NCAM/mg total protein at birth to postnatal day 40 where a level of 0.21 microgram NCAM/mg total protein was found. During aging, NCAM increased again to 0.34 micrograms NCAM/mg total protein at postnatal day 730. NCAM isoforms were studied by immunoblotting analysis. At all ages, the major NCAM isoforms of 190, 145 and 120 kDa were observed in heart, the 145 kDa isoform being predominant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223695 TI - Substrate-bound GP130/F11 will promote neurite outgrowth: evidence for a cell surface receptor. AB - GP130/F11 is a neuronal cell adhesion molecule belonging to the Ig superfamily. Preliminary evidence suggests that it may be important for the fasciculation of axons and cell-cell interactions during synaptogenesis. Previous work has shown that when F3 (the mouse homologue) is transfected into Chinese hamster ovary (CHO) cells it will act as a substrate for neuronal growth and adhesion. In this study we have shown that isolated GP130/F11 will also act as a substrate for the growth of chick dorsal root ganglia cell neurons. The concentration of GP130/F11 on the surface of sympathetic neurons, grown for one day in culture was approximately 40 ng cm-2 which was comparable to the levels required to stimulate neurite outgrowth. Deglycosylation of GP130/F11 or partial denaturation by boiling did not abolish the activity. However, after treatment with dithiothreitol, GP130/F11 was no longer capable of stimulating neurite outgrowth. GP130/F11 will also stimulate neurite outgrowth from forebrain neurons from 7 to 9-day-old embryos. These do not express GP130/F11 until around day 10, providing further evidence that GP130/F11 interacts with an as yet unknown cell surface receptor. PMID- 8223696 TI - Normal rat uterine stromal cells in continuous culture: characterization and progestin regulation of growth. AB - Stromal cells were isolated from rat uterus by sequential enzymatic digestion and density fractionation on Percoll gradient and subcultured by trypsinization. Two stable subcultures, named UII and UIII, were obtained. UII cells exhibited a spindle-shaped, elongated, fibroblast-like morphology, while UIII cells were rounded and polygonal. Both cell types expressed the intermediate filament vimentin but not cytokeratin, nor desmin, suggesting that both were of stromal origin. In UIII cells, the presence of progesterone and prolactin (PRL) receptors was demonstrated by immunocytochemical and binding studies. Cross-linking and Western blotting showed that PRL receptor in UIII cells corresponded to 3 molecular forms of 54, 42 and 32 kDa. The growth properties of these cells were studied under different conditions of culture. In fetal calf serum (FCS) supplemented medium, proliferation of UIII cells was dependent on serum concentration and was not affected by estradiol and progesterone. In 10% FCS supplemented medium, the doubling time was 41.5 +/- 0.8 h. When cultured in 10% dextran-charcoal-treated FCS, cells were maintained in a viable but quiescent state. Under these conditions, progesterone was able to induce growth of these cells in a dose-dependent manner. A 3-fold increase in DNA content was measurable in 10(-7) M progesterone-treated versus control cultures after 5 days. Reduction of serum concentration from 10% to 2% abolished the effect of progesterone suggesting that this effect requires the presence of serum factor(s). In conclusion, this study showed that uterine stromal cells, in continuous culture, retained progesterone and prolactin receptors and progesterone regulation of growth. PMID- 8223697 TI - Thrombin stimulates fibroblast chemotaxis and replication. AB - The serine protease alpha-thrombin, a product of the circulating zymogen prothrombin, plays multiple roles in homeostasis and coagulation. During blood clotting, it is present within fibrin matrices and is likely to be presented to local cell populations. It is known to be a fibroblast mitogen, but its effects on fibroblast recruitment have not been assessed. Here we compared the effect of human alpha-thrombin on chemotaxis and proliferation of human and rat skin fibroblasts and assessed the mechanism of these actions. Fibroblast chemotaxis was assayed using a 48-well Boyden chamber and replication assessed by a spectrophotometric method, based upon the uptake and subsequent elution of methylene blue by fibroblasts. Two fibroblast cell lines were used; fetal rat skin (FR) and newborn human foreskin (HS68). Human alpha-thrombin stimulated FR fibroblast chemotaxis over a wide range of doses (10(-12) M to 10(-7) M). Maximal migration was seen at 10(-10) M; 39 +/- 2.5 cells/high power field (h.p.f.) compared with 19 +/- 3 cells/h.p.f. for media control. In the same assay platelet derived growth factor, a well characterized fibroblast chemoattractant, caused a maximal stimulation of 44 +/- 5 cells/h.p.f. at a concentration of 3 x 10(-9) M. A similar stimulation was observed with HS68 fibroblasts, although for this cell line maximal chemotaxis (190 +/- 12.5% of control) was seen at 10(-8) M thrombin. Fibroblast replication was optimal at 1.25 x 10(-9) M thrombin (134 +/- 4 and 127 +/- 5% of control for FR and HS68, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223698 TI - Autocrine, endocrine and paracrine regulation of growth abnormalities in autosomal dominant polycystic kidney disease. AB - Genetic polycystic kidney disease (ADPKD) is an autosomal dominant trait in man, the phenotypic expression of which is characterized by enormous cystic enlargement of renal tubules. Since this is, in part, a result of aberrant epithelial cell proliferation, the nature of this defect has been characterized by comparison of growth factor action on defined epithelial primary monolayer cultures derived from individually microdissected normal human renal proximal and distal tubules and ADPKD cyst-lining epithelia. Mitogenic assays showed an increased sensitivity of ADPKD epithelia to stimulation by the combination of the endocrine factors hydrocortisone (dexamethasone) and insulin, and Northern analysis suggested increased levels of insulin receptor steady state mRNA. The most potent, single mitogen was epidermal growth factor (EGF), and hypersensitivity to EGF in ADPKD epithelia was correlated with increased mRNA levels for EGF receptor in ADPKD kidneys. The localization in vivo of EGF immunoreactivity in ADPKD cyst-lining epithelia and in (apical) cyst fluids and the demonstration of EGF-receptor immunostaining and specific [125I]EGF binding to apical cell surfaces suggested an autocrine mechanism of growth stimulation by EGF in ADPKD epithelia. Transforming growth factor beta was an inhibitor of normal renal tubule proliferation but was unable to completely inhibit EGF stimulation in ADPKD cultures. Platelet-derived growth factor (PDGF) immunoreactivity which was also seen in ADPKD cyst epithelia, was not mitogenic to ADPKD epithelia but did stimulate division in ADPKD fibroblasts in vitro. This suggested a paracrine regulation of the observed fibrosis in ADPKD. PMID- 8223699 TI - Location of gp330/alpha 2-m receptor-associated protein (alpha 2-MRAP) and its binding sites in kidney: distribution of endogenous alpha 2-MRAP is modified by tissue processing. AB - The alpha 2-macroglobulin receptor-associated protein (alpha 2-MRAP) is a 39 to 44 kDa protein that copurifies with the alpha 2-macroglobulin receptor (alpha 2 MR/LRP) and also with gp330, a highly glycosylated protein located within kidney proximal tubules and glomerular podocytes. Both gp330 and the alpha 2 macroglobulin receptor are members of the low density lipoprotein receptor family but the physiological ligands for gp330 are unknown. In order to understand potential functions of the alpha 2-MRAP, specific anti-alpha 2-MRAP antibodies were used for immunocytochemical studies on paraformaldehyde lysine periodate (PLP)-fixed rat kidneys and on snap-frozen/acetone-fixed tissue. Conflicting results were obtained. After PLP fixation, alpha 2-MRAP was detected almost exclusively in rough endoplasmic reticulum (RER) cisternae; cell surface staining was virtually absent. In snap-frozen tissue, intense staining of the proximal tubule brush border was found, with little or no cytoplasmic staining. A series of experiments showed that during incubation of snap-frozen tissues, endogenous alpha 2-MRAP is released in soluble form from its intracellular location (i.e., the RER) and binds to gp330 on the brush border of proximal tubules. The location of binding sites for alpha 2-MRAP in rat kidney was also examined, using an alpha 2-MRAP-IgG fusion protein. In both snap-frozen and PLP-fixed tissues, this probe bound exclusively to brush borders, and not to intracellular sites. Our results demonstrate: a) that in renal proximal tubule cells, alpha 2-MRAP is located predominantly in the RER, b) that alpha 2-MRAP-binding sites are present on gp330, which is on the proximal tubule brush border, and c) that the apparent brush border localization of alpha 2-MRAP detected in snap-frozen sections is due to an artifactual redistribution of endogenous alpha 2-MRAP that occurs during tissue processing. PMID- 8223700 TI - GEP31, a new gastric epithelial protein, early expressed during ontogenesis. AB - A set of monoclonal antibodies directed against various gastric markers was produced in order to study the developmental expression of the gastric mucosa. A previously described monoclonal antibody, mab 146.14, labeled the proton pump (H+, K+) ATPase specifically located in gastric parietal cells. Mabs 15.1 and 121.17 revealed the mucus of mucous neck cells and mucous surface cells, respectively. By addition, mab 81.1 was directed against a cell surface membrane protein antigen composed of a major 31 kDa component (called GEP31). Our study mainly focused on the characterization of GEP31. This protein was typically located in the gastrointestinal epithelial tract (stomach, small intestine, colon). Moreover, interesting features were observed during the study of the early ontogenesis of the gastric mucosa. The 31 kDa protein was detected at the onset of gland formation (day 18), and a gradual increase in expression of the protein could be observed between day 18 and day 19. Furthermore, a comparative study of the expression of different terminal differentiation markers of gastric epithelial cells ((H+, K+) ATPase, mucins) during the early period of ontogenesis revealed that GEP31 could be detected well before the appearance of these markers. To our knowledge, GEP31 thus appears as the earliest expressed specific cell surface epithelial gastric antigen described to date. Furthermore, the partial N-terminal amino acid sequence of GEP31 was determined and revealed that it is not a known protein. PMID- 8223701 TI - The role of the microtubular cytoskeleton in determining nuclear chromatin structure and passage of maize root cells through the cell cycle. AB - Depolymerization of microtubules in metabolically inactive quiescent center (QC) cells of maize root apices by means of three different antimicrotubular treatments (colchicine, oryzalin and low temperature) elicited very similar responses in their nuclei. Conspicuous nuclear enlargement was closely associated with chromatin decondensation and accelerated traverse of their cell cycle. This latter finding was inferred not only from cytophotometry which showed an increased proportion of S and G2 nuclei in this group of cells, but also from autoradiography which confirmed the greater proportion of nuclei engaged in the S phase of the cell cycle. Activation of the QC cells with various antimicrotubular agents may be a reflection of a dependency of nuclear cell cycle events on the turnover of cytoplasmic microtubules during interphase. The nuclear size, nuclear chromatin structure, as well as cell cycle progression, seem to be regulated by the dynamic nature of the microtubular cytoskeleton. PMID- 8223702 TI - Identification of proteins immunologically related to vertebrate lamins in the nuclear matrix of the myxomycete Physarum polycephalum. AB - Lamin-like proteins have been identified as components of the nuclear matrix of the myxomycete Physarum polycephalum. A 67 kDa homologue was detected by immunoblotting of nuclear matrix proteins with affinity-purified anti-lamin antibodies of human autoimmune sera. A 65 kDa lamin B homologue was identified with polyclonal antibodies against turkey erythrocyte lamin B in nuclei and nuclear matrix of Physarum amoebae and plasmodia. Indirect immunofluorescence microscopy revealed that the 65 and 67 kDa proteins are localized in the nuclear lamina of plasmodial nuclei. PMID- 8223703 TI - A coat protein required for transcytotic traffic exists as a multimeric complex. PMID- 8223704 TI - The DNA topoisomerase I inhibitor camptothecin blocks postmitotic reformation of nucleoli in mammalian cells. PMID- 8223705 TI - Glycosylation in Golgi apparatus of early spermatids of rat. A high resolution lectin cytochemical study. AB - In the present study, lectin cytochemistry in combination with enzyme and chemical treatments and ultrastructural immunocytochemistry were applied to investigate the formation of acrosomal glycoproteins in endoplasmic reticulum (ER) and Golgi apparatus (GA) of early rat spermatids. In addition, the vesicles involved in glycoprotein traffic were investigated using a monoclonal antibody against clathrin. The results obtained suggest the occurrence of high mannose and complex type N-linked oligosaccharides and mucin type O-linked oligosaccharides. In N-linked glycoproteins, Man residues are incorporated into the nascent oligosaccharide in the ER, Fuc residues of the inner core of the oligosaccharide in the cis region of GA, GlcNAc in medial cisternae of GA and Gal residues in the transmost cisternae of GA. In O-linked glycoproteins, the addition of GalNAc occurs in cis and trans cisternae of GA. Gal beta 1,3GalNAc sequence was detected in medial and trans cisternae of GA. Sialic acid was detected in both N- and O linked oligosaccharides in medial and trans cisternae of GA but not in acrosomes. Immunoreactivity to clathrin was observed in the intermediate zone between ER and GA and in vesicles of the trans side of GA. PMID- 8223706 TI - Properties of acidified compartments in hippocampal neurons. AB - We have studied cultured rat hippocampal neurons using electron microscopic procedures based on the accumulation of DAMP (dinitroanilino-aminomethyl dipropylamine) to identify acidified locales, fluorescence procedures to provide information about the pHs within certain endocytic compartments, and the uptake of horseradish peroxidase to evaluate effects of altering pH on membrane cycling in the axonal varicosities. We find that the endocytic compartments related to the lysosome-endosome system in the cell bodies and dendrites of these neurons maintain pHs in the range of about 5 to 6.5. This is the range that would be expected for structures participating in lysosomal digestion and for such functions as the endosomal dissociation of ligands from receptors. We also find that, as judged by uptake of horseradish peroxidase, exposure of the preparations to weak bases that neutralize intracellular compartments does not abolish the endocytic labeling of synaptic vesicles in the axonal varicosities. This suggests that passage through a markedly acidified compartment or stage is not obligatory for the endocytic phase of the cycling of synaptic vesicles. PMID- 8223707 TI - Circular ruffle formation and closure lead to macropinocytosis in hepatocyte growth factor/scatter factor-treated cells. AB - Treatment with hepatocyte growth factor/scatter factor (HGF/SF) rapidly induced the formation of conspicuous circular ruffles on the apical surfaces of two kidney cell lines, MDCK and PtK2. The ruffles were found to contain significant amounts of F-actin and myosin as judged by immunofluorescence microscopy. Time lapse photomicroscopy demonstrated that the ruffles constrict, closing over, and were followed by the formation of phase bright structures. That these structures were macropinocytotic vesicles was confirmed using fluorescein isothiocyanate (FITC)-dextran as a marker for fluid uptake. It is hypothesized that the constriction of the ruffles followed by membrane fusion causes the vesicles to form. Treatment with suramin blocked both circular ruffle formation and scattering, suggesting that ligand binding was the causal agent for ruffle formation. The drugs amiloride and SITS (4-acetamido-4'-isothiocyanatostilbene 2,2'-disulfonic acid) also completely inhibited ruffle formation, suggesting that ion transport was an early consequence of HGF/SF binding and that these transport effects had a major role in the cytoskeletal changes leading to circular ruffle formation. The appearance of macropinocytotic vesicles was also blocked by amiloride treatment. Surprisingly though, subsequent scattering was not blocked by amiloride treatment, although suramin and SITS both entirely inhibited scattering. PMID- 8223708 TI - Autophagic vacuoles fuse with the prelysosomal compartment in cultured rat fibroblasts. AB - We previously demonstrated both mannose 6-phosphate receptor (MPR) and cathepsin L in early autophagic vacuoles of cultured rat fibroblasts. This suggested that the enzyme may originate either from the receptor-enriched prelysosomal compartment (PLC) or from the trans-Golgi network (TGN). In the present ultrastructural study, we elucidated the roles of the PLC and TGN in lysosomal enzyme delivery to autophagic vacuoles. Firstly, we studied whether endocytic markers, cationized ferritin (CF), bovine serum albumin-gold or horseradish peroxidase (HRP), can be detected in autophagic vacuoles. Autophagy was induced by serum removal from the medium with or without leupeptin, an inhibitor of cysteine proteinases. Endocytic markers were not detected in autophagic vacuoles after short uptake which filled the early endosome, but only after longer labeling which filled the PLC. The markers were usually found in advanced autophagic vacuoles containing partially degraded cytoplasm and complex internal membranes which are the characteristic of the PLC. HRP-positive vesicles were also observed in continuity with early autophagic vacuoles containing intact cytoplasm. After uptake and transport of CF and HRP to the PLC, these markers were delivered to autophagic vacuoles even if microtubules were disrupted in vinblastine before the induction of autophagy. Secondly, we studied whether MPRs transport cathepsin L to autophagic vacuoles directly from the TGN. Two inhibitors of MPR-mediated enzyme transport, tunicamycin and chloroquine, were used. Quantitative immunocytochemistry showed that neither of these drugs prevented cathepsin L delivery to autophagic vacuoles. The results suggest that a large proportion of lysosomal enzymes is delivered to autophagic vacuoles from the PLC by a microtubule-independent manner. The first enzymes may be transported in small PLC-derived vesicles or tubules which are reached by HRP but not by CF and gold. Later, the autophaged cytoplasm is delivered to larger vacuolar parts of the PLC. Mannose 6-phosphate receptors transport no or only trace amounts of lysosomal enzymes to autophagic vacuoles directly from the TGN. PMID- 8223709 TI - Formation of autophagosomes during degradation of excess peroxisomes induced by administration of dioctyl phthalate. AB - The formation of autophagosomes in rat hepatocytes was investigated during degradation of excess peroxisomes. Rat liver peroxisomes were markedly proliferated by administration of dioctyl phthalate (DEHP) for 2 weeks. When the animals were fed on normal diet for a week further, the number and size of the peroxisomes recovered to normal. The recovery process was confirmed by the assay and immunoblot analysis of acyl-CoA oxidase and catalase. During the recovery process, only a few autophagosomes were noted. However, when leupeptin (2 mg/100 g body weight) was injected into these animals, there was a marked accumulation of autophagosomes in the hepatocytes. Using this as an experimental model, the early stage of the autophagosome formation was analyzed by electron microscopy. Twenty minutes after the injection, isolation membranes surrounding the target organelles appeared. They were characterized by double layers with a narrow cisternal space and were sometimes continuous with the rough endoplasmic reticulum. Between the inner membrane of the isolation membranes and the enclosed organelles, electron-dense bridges were noted. Forty minutes after leupeptin injection, the lumen of the isolation membranes were enlarged and the inner membrane attached to the entrapped material. Enzyme cytochemical staining showed that the isolation membranes were negative for acid phosphatase and thiamine pyrophosphatase, but were strongly positive for glucose-6-phosphatase (G6Pase). The enlarged cisternae of the isolation membranes of the early autophagic vacuoles were in part positive for this enzyme, but gradually became negative with time. Similarly, the G6Pase activity was lost when the inner membrane was degraded. The results suggest 1) that the process of degradation of excess peroxisomes is rapid and carried out by the autophagic system in hepatocytes and 2) that the isolation membranes enclosing the target organelles are derived from the endoplasmic reticulum. PMID- 8223710 TI - Evidence for the presence of proteolytic activity in peroxisomes. AB - The presence of endo- and exoproteolytic activity in peroxisomes was detected in cell organelles purified from pea leaves. By PAGE using different exopeptidase substrates (L-aa-beta NA), one leucine aminopeptidase (AP) was found in peroxisomes. The peroxisomal AP was characterized as a serine protease and had a maximal activity at pH 7.5, a molecular mass of 56.8 kDa and a pI of 5.3. This enzyme was mainly present in the soluble fraction of peroxisomes. The occurrence of proteases in peroxisomes suggests that they might be involved in the protein turnover and processing of imported precursor polypeptides in peroxisomes. PMID- 8223711 TI - The hydrogenosomal enzyme hydrogenase from the anaerobic fungus Neocallimastix sp. L2 is recognized by antibodies, directed against the C-terminal microbody protein targeting signal SKL. AB - The question was addressed whether antibodies directed against the general microbody C-terminal protein targeting signal SKL recognized hydrogenosomal proteins from Neocallimastix sp. L2. Immunofluorescence, immunocytochemistry and Western blotting experiments using these antibodies indicated the presence of hydrogenosomal proteins containing SKL-COOH. One of these proteins, the hydrogenase, was purified to homogeneity. It has a native molecular mass of 87 kDa and consists of two subunits of approximately 30 and 60 kDa, both cross reacting with anti-SKL antibodies. Its activity could be inhibited by CO, NO2-, and acetylene, suggesting a (Ni-Fe-Se) hydrogenase. Immunocytochemistry using polyclonal antibodies raised against the hydrogenase revealed the location of this protein in the hydrogenosomal matrix. The results described in this paper suggest that hydrogenosomes from Neocallimastix sp. L2 are related to microbodies from aerobic eukaryotes and support the idea of a common evolutionary origin for these organelles. PMID- 8223712 TI - Multivesicular bodies in HEp-2 cells are maturing endosomes. AB - Conventional fluorescence microscopy of fixed HEp-2 cells as well as video microscopy of living cells incubated with transferrin-Texas Red (Tf-TxR) for < 60 min revealed distinct punctuate endosomal structures. Quantitative ultrastructural analysis using horseradish peroxidase (HRP) and cationized gold as tracers showed that spherical multivesicular bodies (MVBs) were the predominant endocytic compartments in HEp-2 cells and that MVBs within 60 to 90 min matured into lysosomes still containing internal vesicles. The number of labeled MVBs increased continuously from 2.5 min to 30 min of tracer incubation. However, when the cells were pulsed for 5 min followed by 10 or 25 min chases, the number of labeled MVBs corresponded to that obtained after 5 min of continuous incubation. The diameter of labeled MVBs was largely constant with time, but the number of internal MVB vesicles increased. Thus, early or newly formed MVBs contained few internal vesicles, whereas late MVBs, that is to say, MVBs that have existed for some period of time, contained numerous internal vesicles, and finally a mixture of membranous material or myelin figures and vesicles. It is thus in principle possible to distinguish between early and late MVBs in HEp-2 cells on the basis of morphology. However, the difference in number of internal vesicles applies only to the entire MVB population; after only 2.5 to 5 min of incubation, MVBs with numerous internal vesicles could also be reached by internalized tracer. Concomitant with the gradual changes in morphology, the MVBs also showed a characteristic change in content of marker proteins as detected by immunogold labeling on ultracryosections. Hence, early MVBs with relatively few internal vesicles and typically reached by internalized tracers within 5 min contained transferrin receptors (TfRs). By contrast, MVBs with many internal vesicles and labeled after 60 min of incubation contained mannose phosphate receptors (MPRs), and the MVBs with distinct membranous material or myelin figures in addition to the internal vesicles were enriched in the lysosome membrane protein lamp-1. Thus, there seems to be a gradual maturation of MVBs in HEp-2 cells. PMID- 8223713 TI - Reversible pH-induced homophilic binding of GP2, a glycosyl-phosphatidylinositol anchored protein in pancreatic zymogen granule membranes. AB - GP2, the major zymogen granule membrane (ZGM) protein in the pancreas, is linked to the lumenal leaflet of the lipid bilayer via a glycosyl-phosphatidylinositol (GPI) moiety. We demonstrate that the peptide domain of GP2 (pGP2, approximately 75 kDa), purified from pancreatic ZGMs after phospholipase C cleavage, shows pH- and calcium-dependent self-association into sedimenting complexes. This homophilic binding process is progressive as pH is reduced from 7.0 to 5.5 and calcium is increased from 0 to 10-20 mM. This self-association reaction is temperature-dependent, optimal between 20 and 37 degrees C, progressively reduced below 20 degrees C, and eliminated at 10 degrees C. The reaction is reversible as a function of pH and abolished in the presence of nonionic detergents. Specificity in the homophilic reaction is demonstrated by the exclusion of heterologous proteins (globin, serum albumin, and IgG) from sedimenting complexes. At pH 5.5 in the presence of 20 mM calcium, oligomeric structures (approximately 300 kDa) consistent with tetrameric complexes were observed by gel filtration chromatography and elliptical structures (14-18 nm), frequently arranged in variegated clusters, were observed in the electron microscope by negative staining techniques. The pH- and calcium-dependent self-association observed for GP2 may represent an important mechanism by which GPI-anchored membrane proteins engage in homotypic binding reactions to establish highly functional membrane (micro)- domains targeted to regulated secretory compartments in polarized epithelial cells. PMID- 8223714 TI - Defective secretion of albumin encoded by exon-skipped mRNA found in aged analbuminemic rat liver. AB - Remarkable increases in the amount of exon H-skipped (delta H), exons G and H skipped (delta GH), and exons H and I-skipped (delta HI) mRNAs of rat albumin have been found in the livers of aged or mutagen-treated analbuminemic rats (Kaneko, T., H. Shima, H. Esumi, M. Ochiai, S. Nagase, T. Sugimura, M. Nagao: Proc. Natl. Acad. Sci. USA 98, 2707-2711 (1991)), in association with increases in numbers of immunohistochemically albumin-positive hepatocytes and the accumulation of albumin(s) in the cells. To determine which type of mRNA is responsible for the accumulation of albumin(s), expression vector systems for intact mRNA and three kinds of exon-skipped albumin mRNAs were constructed and transfected into COS-1 and HepG2 cells. Transient expression and secretion of albumins were examined in these cells. delta HI albumin and intact albumin were efficiently expressed by both types of transfected cells. The delta HI albumin expressed had a similar molecular weight to the major albumin isolated from the microsome fraction of the liver of aged analbuminemic rats, as judged by SDS PAGE. The rate of secretion of delta HI albumin was very low in both types of transfected cells, whereas those of delta H and delta GH albumins were normal. Immunocytochemical analyses of the transfected COS-1 cells with antiserum against rat albumin showed that the distribution of intact albumin had a compact Golgi like pattern, whereas that of delta HI albumin had a diffuse endoplasmic reticulum-like pattern. These distribution patterns were similar to those in aged analbuminemic rat hepatocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223715 TI - Brefeldin A effects on tobacco pollen tubes. AB - Pollen germination and pollen tube growth were clearly affected by the drug brefeldin A: both processes could be stopped completely. Notable thickening of the cell wall was not observed in non-growing pollen tubes, and plasma streaming remained vigorous in the presence of brefeldin A. Over time, the tip region became filled with a dense cytoplasm that lacked large organelles. Ultrastructural observation showed the rapid dissociation and disappearance of Golgi bodies. The concomitant appearance of vesicle-like structures attached to the endoplasmic reticulum (ER) suggests a fusion of the Golgi with the ER similar to that which happens in animal cells. Secretory vesicles disappeared from drug treated tubes, and the tip region became filled with tubular ER. As the actions of the drug seemed to be reversible, brefeldin A may contribute substantially to research on Golgi dynamics and tip growth in pollen tubes. PMID- 8223716 TI - Colocalization of low- and high-affinity NGF receptors on PC12 cells, C6 glioma cells and dorsal root ganglion neurons. AB - The biological responsiveness of neural cells to nerve growth factor (NGF) appears to require expression and ligand binding to both the low-affinity NGF receptor (LNGFR) and the proto-oncogene product trk, the latter being a receptor tyrosine kinase. Immunolocalization of the LNGFR and the high-affinity component of the NGF receptor, trk (HNGFR) was studied by electron microscopic morphometric analysis on cultured PC12 pheochromocytoma cells, C6 glioma cells and neonatal rat dorsal root ganglia neurons using a double immunogold labeling technique. Two receptor-specific antibodies, anti-LNGFR monoclonal antibody 192-IgG and a polyclonal antibody against the 14 carboxy-terminal amino acids of the Trk protein, were utilized in conjunction with immunoglobulin conjugated to colloidal gold particles of different sizes. All cells treated with NGF (50 ng/ml) displayed significant colocalization of LNGFR/HNGFR-like immunoreactivity. Gold particles associated with LNGFR (LNGFR-like immunoreactivity) were frequently seen near 2 or 3 (or more) particles delineating the HNGFR on all cell surfaces. Positive Trk-like immunoreactivity (HNGFR) thus seems to localize in close proximity to LNGFRs in at least these cell types. PMID- 8223717 TI - Localization of the CHIP28 water channel in reabsorptive segments of the rat male reproductive tract. AB - The water channel protein CHIP28 is responsible for the high constitutive plasma membrane permeability to water of erythrocytes, renal proximal tubule, and thin descending limb of Henle. The male reproductive tract is embryologically related to kidney and some segments, particularly the efferent ducts, exhibit a high rate of solute flux-dependent reabsorption of luminal fluid. To determine whether this could occur through water channels in the plasma membrane of reproductive tract epithelial cells, we used anti-CHIP28 antibodies to localize this protein by Western blotting and immunocytochemistry. Western blotting of proteins from efferent duct homogenate indicated the presence of CHIP28 in the efferent duct cells. By indirect immunofluorescence and protein A-gold immunolabeling, CHIP28 was localized to the brush-border and basolateral membranes of nonciliated cells. Ciliated cells in the same epithelium showed no plasma membrane staining for CHIP28. In accord with immunocytochemical findings, freeze-fracture of nonciliated efferent duct cells revealed a plasma membrane organization resembling that of renal proximal tubule cells that are rich in CHIP28. The anti CHIP28 antibodies also stained plasma membranes of epithelial cells in the ampulla of the vas deferens, seminal vesicles, and prostate, but not the cells in seminiferous tubules, epididymis, and proximal parts of the vas deferens. Therefore, CHIP28 may be a principal mediator of the transmembrane water transport in absorptive epithelial cells of efferent ducts, as well as in epithelial of several other segments of the male reproductive tract that show both secretory and reabsorptive functions. PMID- 8223718 TI - Complexus adhaerentes, a new group of desmoplakin-containing junctions in endothelial cells: the syndesmos connecting retothelial cells of lymph nodes. AB - We describe, in bovine and human tissues, a new kind of adhering junctions ("complexus adhaerens") which occur in certain vascular endothelial and, by morphological and compositional criteria, are neither desmosomes nor adhering junctions of the zonula (fascia) adhaerens category. A particularly abundant and complex subtype among the "complexus adhaerens" junctions is characteristic of the endothelium-related, stellate cells that form the three-dimensional filter meshwork of lymph node sinus. These "retothelial" cells are multiply interconnected by junctions which are highly polymorphic in shape and size, often show interruptions and branches and can be traced for several micrometers. Their dense cytoplasmic plaque contains plakoglobin and desmoplakin but is practically negative for vinculin and alpha-actinin. In contrast to true desmosomes, including those of the dendritic reticulum cells of lymph node follicles, the retothelial junctions are negative for any of the known desmosomal cadherins (desmogleins and desmocollins) and also for E- and M-cadherin. These cells, however, are rich in cadherin-5 which colocalizes with desmoplakin and plakoglobin in numerous, though not all sites, indicating that here cadherin-5 may contribute to the formation of a desmoplakin plaque. The complex arrangement of the retothelial form of complexus adhaerens, for which the term "syndesmos" is proposed, has been reconstructed from electron micrographs by a computer-aided design. We also report that desmoplakin-positive complexus adhaerens junctions constitutively occur in various lymphatic endothelia of many other tissues, most prominently in submucosal lymph vessels, and discuss the possible function of this kind of junctions as a means to fortify cell connections between the tenuous endothelial cells and, in the lymph node sinus, to support their filtering functions. PMID- 8223719 TI - Thrombomodulin and thrombin localization on the vascular endothelium; their internalization and transcytosis by plasmalemmal vesicles. AB - We have immunolocalized thrombomodulin (TM), (an endothelial cofactor protein for thrombin-mediated protein C activation), and its ligand thrombin (TH) by light- and electron microscopy on endothelial cells in culture and in situ. Our results show that: i) TM is expressed in small clusters on the basal and the apical surface of human umbilical vein endothelial cells (HUVEC), is absent from coated pits and coated vesicles, is internalized from the cell surface via plasmalemmal vesicles, and is associated with endosomes, including a tubulovesicular network within the cytoplasm; ii) exogenous TH binds at low surface density on HUVEC and colocalizes with TM in small TM clusters; iii) on the microvascular endothelium of heart, lung, and kidney, TM is expressed at high density on the luminal plasmalemma, is associated with a fraction of the plasmalemmal vesicle population; iv) and is found internalized in multivesicular bodies in TH perfused heart samples; v) in the same samples, perfused TH is detected at low surface density: on the luminal aspect of the endothelium within small TM clusters, in the introits and within plasmalemmal vesicles, in endosomal structures, on the abluminal plasmalemma, and within the pericapillary spaces. Our findings suggest that the high level of endothelial TM expression in situ provides a safety margin for random TH generation; they also demonstrate that bound TH is transcytosed across the endothelium by plasmalemmal vesicles which might represent an additional mechanism to remove TH from the circulation. PMID- 8223720 TI - Thymosin beta 4 (T beta 4) in activated platelets. AB - When resting human blood platelets are stimulated with thrombin, 50 to 60% of the G-actin polymerizes to F-actin within 60 seconds. The increase in F-actin is correlated with a corresponding decrease in the complex of G-actin with T beta 4. Within 5 seconds after stimulation, nucleation sites for pyrene actin polymerization increase 1.5 times in Triton-lysed supernatants. Cytochalasin D, known to inhibit the increase in F-actin after thrombin, also inhibits the fall in T beta 4-actin complex and the increase in nucleation sites. Phosphorylation of T beta 4 could not be detected in either control or activated cells. Increased T beta 4 corresponding to that lost from the T beta 4-actin complex is present in lysates from activated platelets and retains the ability to complex with actin. The data, taken together with previous estimates for the dissociation constant of the T beta 4-actin complex, show that actin polymerization following platelet activation could be controlled primarily by the increased availability of free barbed ends of actin filaments which have a higher affinity for G-actin than does T beta 4 and suggest that the increased free T beta 4 may serve to limit the degree of polymerization. PMID- 8223721 TI - Galactose-specific receptor modulation related to the onset of apoptosis in rat liver. AB - The expression of the asialoglycoprotein receptor of hepatocytes and the galactose-specific receptors of non-parenchymal liver cells during the onset of apoptosis in liver of rats treated with lead nitrate was studied. During the involution of lead nitrate-induced hyperplasia in rat liver (occurring at 5 days after the injection) a significant increase of asialoglycoprotein receptor (ASGP R) expression on hepatocytes coincided with the massive death by apoptosis of the same cells. The increase in the receptor expression was sustained by a large increase in the level of its specific mRNA. As a consequence of lead nitrate injection, we also detected a drastic change of the galactose-specific receptor expression and distribution on the surface of rat liver sinusoidal cells. However, the modulation of the receptor expression on the Kupffer cells did not parallel that observed for the ASGP-R: the peak of surface expression measured on hepatocytes always followed the one observed on Kupffer cells. Our data show a first evidence of a receptor modulation during the process of apoptosis. In fact, the entire carbohydrate recognition system of the liver is modulated during the onset of apoptosis induced by lead nitrate injection, but the pattern of modulation depends on the cellular types. We suggest that a physiological role for the hepatic carbohydrate recognition systems is related to the apoptosis of liver. PMID- 8223722 TI - Three-dimensional ultrastructure of generative cell mitosis in the pollen tube of Nicotiana tabacum. AB - Generative cell mitosis was examined in stylar-grown pollen tubes of Nicotiana tabacum using serial sectioning, transmission electron microscopy and computer assisted reconstruction. Before mitosis, the generative cell has a cage-like organization of cytoplasmic microtubules. The mitotic spindle forms when the cytoplasmic microtubules reduce in frequency and kinetochore microtubules form in an area delimited by sheets of endoplasmic reticulum; no preprophase band of microtubules is observed. At metaphase, 21 pairs of kinetochores are distributed unevenly along the length and depth of the cell without the formation of a strictly planar metaphase plate. The metaphase spindle is highly oblique, with diffuse subpoles distributed along the sides of the cell, colocalized with endoplasmic reticulum lamellae. From these dispersed subpoles the kinetochore bundles emanate, closely associated with tubular endoplasmic reticulum. Anaphase consists of three principal processes: convergence of diffused mitotic poles, shortening of the kinetochore bundles, and the elongation of the spindle by an average of nearly 50%. At mid-anaphase, a phragmoplast begins to form, mainly by the assembly of new microtubules at the equatorial area, which form as a cluster of numerous short microtubules. Cytokinesis is essentially conventional, with centrifugal cell plate formation. Cytoplasmic microtubules are restored in the newly formed "brother" sperm cells in a distribution similar to that in the generative cell but fewer in number. PMID- 8223723 TI - Characterization of the interchromatin region as the nuclear domain containing snRNPs in plant cells. A cytochemical and immunoelectron microscopy study. AB - The combination of electron microscopy (EM) cytochemical with immunocytochemical methods is used to characterize the interchromatin region (IR) of the plant cell nucleus. Cryoprocessing of the sample provides a better ultrastructural preservation and allows the observation of some differences in the fine structure of the IR which shows a denser aspect resulting from the lower extraction of components with low-temperature methods. A complex network of fibrillar structures and isolated or clustered 30 to 50-nm granules are observed in the IR. Anti-DNA antibodies combined with the NAMA-Ur method for DNA or the EDTA staining, preferential for RNPs, allow the detection of chromatin fibers in the IR. Bismuth staining reveals the presence of highly phosphorylated proteins in some interchromatin structures. The spliceosomal snRNPs are immunolocalized on cryosections and Lowicryl sections of plant cells using monoclonal and polyclonal antibodies. They provide a homogeneous immunofluorescence pattern with no speckles. This is in correlation with the labeling at EM, immunogold particles decorate the EDTA-positive fibrillar structures of the IR but no labeling is found over the 30 to 50-nm granules. The presence of the spliceosomal snRNPs, DNA and phosphorylated proteins in the IR indicate that this nuclear domain plays a major role in pre-messenger RNA splicing and, possibly in transcription, in the plant cell nucleus. PMID- 8223724 TI - Requirements for nuclear translocation and nucleolar accumulation of nucleolin of Xenopus laevis. AB - The intracellular localization of germinal vesicle proteins is regulated during early Xenopus development. Here we have analyzed the determinants that control the localization of nucleolin, a nucleolar protein of vertebrates. Our immunological analyses and isolation of cDNAs have revealed the presence of a second nucleolin gene in addition to that previously identified. The two nucleolin polypeptides of 95 kDa and 90 kDa molecular mass are both expressed in oocytes and are found predominantly in the nucleoli. During oocyte maturation, both polypeptides are hyperphosphorylated and found distributed throughout the cytoplasm. Hyperphosphorylated forms and cytoplasmic localization of nucleolin both prevail in early embryos up to the midblastula transition. Subsequently, the proteins are found in the nuclei. Accumulation in nucleoli is only detected at later stages, beginning with gastrulation. Thus nucleolar localization is temporally uncoupled from nuclear translocation. Consistent with these observations, our molecular analyses have revealed that the nuclear location signal which is present in nucleolin and which is sufficient for nuclear location, is not sufficient for nucleolar accumulation. Nucleolar accumulation requires, in addition to the nuclear location signal, the presence of the RNA binding domains and of an RG-rich domain, which is also thought to interact with RNA. PMID- 8223725 TI - Evolution of a cortical collecting tubule primary culture. AB - The evolution of a primary culture of rabbit kidney cortical collecting duct (CCT) was followed with the electron microscope using two monoclonal antibodies directed against the principal (Mab 703) and intercalated (Mab 503) cells, respectively. As a result of the loss of the basement membrane surrounding the seeded tubule, the intercalated cells showed a tendency to be eliminated while the basal cytoplasm of the remaining cells consisting mainly of principal cells, quickly spread out at the surface of the filter. Between the first and the seventh hour, cells underwent rapid processes of both dedifferentiation and redifferentiation. At 48 h and later on, they started to proliferate with the production of many multinucleated cells. Normal mitotic divisions, in contrast, were rarely encountered. Whereas the number of intercalated cells as recognized by Mab 503 increased from the fourth day up to the tenth day corresponding to a fully mature culture, culture cells at all time intervals rather resembled principal cells found in the internal part of the cortex or in the outer stripe of the external medulla. It is suggested that in our experimental conditions, dedifferentiated principal cells give rise to both principal and intercalated cells as recognized by immunocytochemistry in the fully developed cell culture. PMID- 8223726 TI - Mitochondria in cultured human muscle cells depleted of mitochondrial DNA. AB - Cultured human muscle cells were depleted of mitochondrial DNA (mtDNA) by prolonged treatment with ethidium bromide (EB). In these respiration-deficient muscle cells neither cytochrome c oxidase activity nor mtDNA were detectable. However, mitochondrial matrix enzymes remained present and were localized in mitochondria-like organelles, as shown by subcellular fractionation. Metabolic labeling showed synthesis of cytochrome c oxidase subunits coded by nuclear DNA (nDNA). These results indicate that depletion of mtDNA in cultured human myoblasts does not inhibit expression of nDNA-coded mitochondrial proteins. The characteristic thread-like pattern of mitochondria was lost in mtDNA-depleted myoblasts, as shown by immunofluorescence with antibodies against cytochrome c oxidase and the F1 part of the mitochondrial ATP synthase (F1-ATPase) and by fluorescence of the carbocyanine dye, 3,3'-dipentyloxacarbocyanine iodide (DiOC5(3)). The organelles visualized by these methods were round and swollen and had a localization different from lysosomes as shown by double-labeling with mitochondrial and lysosomal antibodies. These results indicate that not only synthesis, but also import of mitochondrial proteins into mitochondria-like organelles remains possible in respiration-deficient cells. PMID- 8223727 TI - Long-term prediction of coronary heart disease mortality in two rural Greek populations. AB - In 1960-61 two pooled Greek rural populations totalling 1215 men aged 40-59 years were followed-up for 25 years. A Cox model analysis of fatal coronary events over 15 years showed that serum cholesterol in men aged 40-59 years, cholesterol in men aged 45-64 years, and systolic blood pressure in men aged 50-69 played a predictive role. The coefficient of age became more significant with advancing age and that of cigarette smoking only at 25 years follow-up. The coefficient of cholesterol decreased stepwise and became negative for men aged 50-69; body mass index was without effect in any follow-up of these cohorts. Systolic blood pressure and serum cholesterol increased in these populations by 5.4 mmHg and 23.5 mg.dl-1 (0.61 mmol.l-1), respectively between the years 0 and 10, whereas cigarette consumption decreased minimally. These changes were used to test the predictability of coronary events occurring between years 10 and 25 of follow-up when added to the model containing the factors at entry. Of these changes only systolic blood pressure significantly increased the predictability of coronary deaths. It is concluded that even minor alterations in systolic blood pressure above or below the entry levels can be associated with marked modifications in coronary mortality above or below those occurring naturally in the 15 years after the changes occurred. PMID- 8223728 TI - Trends in hospitalization for heart failure in Scotland 1980-1990. AB - Heart failure is a growing public health problem in industrialized countries with ageing populations. Scotland has a relatively stable population of approximately 5 million and a well described system for recording details of hospitalizations (Scottish Hospital In-Patients Statistics-SHIPS). We have examined SHIPS data for hospitalizations for heart failure in Scotland 1980-1990. Discharges for heart failure as the primary diagnosis increased by almost 60%, from 1.30 to 2.12/1000 population in this period (as either primary or secondary diagnosis the rate increased from 2.51 to 4.24/1000). Seventy-eight percent of discharges were in persons aged > or = 65 years and 48% of discharges were male. Heart failure (primary diagnosis) accounted for almost 4% of all general (internal) medicine discharges. In-patient case fatality was 18% in 1990. Mean duration of in-patient stay on Internal Medicine wards was approximately 11 days. The number of hospitalizations for heart failure is now almost identical to those for myocardial infarction. These trends mirror those recently reported from the United States. Heart failure is an increasingly common and costly cause of hospitalization in Scotland. Approaches which can reduce this burden on the hospital service require urgent attention. PMID- 8223729 TI - Radionuclide assessment of regional differences in left ventricular wall motion and myocardial perfusion in idiopathic dilated cardiomyopathy. AB - Regional variations in left ventricular contractility and myocardial perfusion are frequent in idiopathic dilated cardiomyopathy and might result from an increase in left ventricular wall stress responsible for regional wall motion abnormalities. The aim of the study was to perform radionuclide studies in patients with idiopathic dilated cardiomyopathy to assess regional left ventricular wall motion and myocardial perfusion abnormalities in this myocardial disease. We studied 29 men referred with idiopathic dilated cardiomyopathy and normal coronary angiograms. Rest radionuclide left ventriculography and exercise thallium-201 tomography were performed in all patients. The thallium-201 tomograms were divided into 20 segments for each patient. Mean left ventricular ejection fraction was 27 +/- 11%; 17 patients had diffuse hypokinesia (mean left ventricular ejection fraction: 24 +/- 9%) and 12 patients had predominant regional hypokinesia (mean left ventricular ejection fraction: 32 +/- 12%). Of all 580 tomographic segments, 186 had a reduction of thallium-201 uptake at exercise. Among them, reversibility was found in 53%. On the whole, 68% (158/232) of anterior, inferior and apical segments had a perfusion abnormality, compared with 8% (28/348) of septal and lateral segments (P < 0.0001). Left ventricular wall motion and myocardial perfusion abnormalities are heterogeneous and not evenly distributed in dilated cardiomyopathy. The alterations are predominant on the myocardial regions delineating the antero-posterior axis of the left ventricle. These findings suggest the possible role of increased left ventricular wall stress on this axis. PMID- 8223730 TI - Ischaemic threshold and haemodynamic changes during angina at rest in patients with unstable angina. AB - To examine whether increases in heart rate might be a common trigger of angina at rest, changes in heart rate, blood pressure and rate-pressure product during pain were compared with the ischaemic threshold (heart rate with ST segment shift > = 1 mm), determined by atrial pacing, in 272 patients with unstable angina. During an average of 5.9 +/- 5.2 episodes of angina, heart rate was comparable to control values (77.0 +/- 14.5 vs 75.2 +/- 11.5, beats.min-1, ns) and significantly lower than the ischaemic threshold (147.9 +/- 22.9, P < 0.00001). The rate-pressure product was also lower (955 +/- 183 vs 2033 +/- 369, x 10, P < 0.00001). Heart rate during rest angina was lower than the ischaemic threshold even when we considered only patients with ST depression during pain (n: 71, 81.4 +/- 16.0 vs 132.8 +/- 21.4, P < 0.00001), those with three-vessel disease (n: 43, 79.9 +/- 15.9 vs 136.9 +/- 22.0, P < 0.00001), or those with a low ischaemic threshold (= < 130 beats.min, n: 78, 77.0 +/- 14.9 vs 118.3 +/- 10.7, P < 0.00001). In 154 patients in whom a second pacing test was performed the response was reproducible in 137 cases (89%). Thus, heart rate barely changes during angina at rest in patients with unstable angina and is consistently much lower than the ischaemic threshold. These findings support the concept that increases in heart rate are an unlikely trigger of ischaemia at rest, even in patients with markedly reduced coronary reserve. PMID- 8223731 TI - Velocity distributions in the left ventricular outflow tract and the aortic anulus measured with Doppler colour flow mapping in normal subjects. AB - This study was designed to investigate the velocity distributions in the left ventricular outflow tract and aortic anulus. In 18 out of 22 healthy male individuals, instantaneous cross-sectional flow velocity profiles were constructed at different levels of the left ventricular outflow tract and aortic anulus by time interpolation of digital velocity data from sequentially delayed Doppler colour flow maps. The results showed that: (1) the velocity distributions in the left ventricular outflow tract and the aortic anulus were skewed with the highest velocities along the anterior and septal parts of the flow channel; (2) based on the time-velocity integral profiles in the aortic anulus, which were also skewed with the highest integrals along the anterior and septal parts, the maximal time-velocity integrals were higher than the mean cross-sectional time velocity integrals by approximately 30% in the four chamber view and 40% in the long axis view. However, the time-velocity integrals at the middle point of the diameter correlated significantly with the mean cross-sectional time-velocity integrals in the four chamber view (10.3 +/- 0.8 vs 9.9 +/- 0.9 cm; r = 0.95) and in the long axis view (12.5 +/- 0.9 vs 11.8 +/- 0.8 cm; r = 0.95). Therefore, it can be concluded that: (1) the velocity distributions in the left ventricular outflow tract and the aortic anulus are skewed; (2) if the aortic anulus is used for cardiac output measurement by pulsed Doppler echocardiography in normal subjects, the middle point of its diameter is the best sampling site. PMID- 8223732 TI - Autonomic imbalance in the recovery period after myocardial infarction. AB - The aim of the study was to evaluate the responses to autonomic function tests during the healing period of myocardial infarction (AMI). In 24 patients, at 2 and 6 weeks after the acute event, Valsalva manoeuvre, deep breathing at 6 breaths per min, isometric handgrip and cold pressor tests were performed. Responses of arterial blood pressure (BP) and heart rate (HR) were measured. At 2 weeks post AMI a significant decrease in parasympathetic tone was noted: HR reduction during deep breathing: 18.5 +/- 5.7 beats.min-1 for controls vs 9.7 +/- 2.6 beats.min-1 for AMI, P < 0.001. Systolic BP response to handgrip and cold pressor test was only slightly (non significantly) increased: 30.6 +/- 12.9 mmHg (controls) vs 40.0 +/- 20.5 mmHg (AMI) for the handgrip test and 13.8 +/- 8.1 mmHg vs 18.0 +/- 10.1 mmHg respectively for the cold pressor test. At 6 weeks post AMI, the response to the deep breathing test (15.9 +/- 5.6 beats.min-1) no longer significantly differed from that in controls. In contrast, the sympathetic stressor tests showed a significant increase in systolic BP response: 63.8 +/- 21.9 mmHg, P < 0.001 and 26.1 +/- 14.9 mmHg, P < 0.05, respectively for the handgrip and cold pressor tests. It appeared that infarct localization had no effect on autonomic function tests. There is evidence of autonomic imbalance both at 2 and 6 weeks after AMI, with a rapid but transient decrease in vagal activity and enhancement of orthosympathetic nervous tone; this was markedly more pronounced at 6 weeks post AMI despite treatment with beta-blocking agents in all patients. PMID- 8223733 TI - Prognostic value of thallium-201 myocardial scintigraphy after atrial transoesophageal pacing in patients with suspected coronary artery disease. AB - Fifty-five patients with suspected coronary artery disease underwent planar thallium-201 myocardial scintigraphy after atrial transoesophageal pacing. Coronary angiography was carried out in all patients. Eighteen patients had no myocardial infarction, but a greater than 50% narrowing of at least one main vessel: initial hypoperfusion with redistribution at 4 h occurred in 16 patients (sensitivity 89%). Twenty-one patients had had a previous myocardial infarction: a reversible thallium defect was observed in 12 patients and an irreversible defect in the nine remaining patients. Sixteen patients had normal coronary arteries: a reversible thallium defect was observed in three patients (specificity 81%). After a mean follow-up of 22 +/- 13 months (range 6 to 40), 23 cardiac events occurred: cardiac death in one patient, unstable angina in three, and revascularization procedures for recurrent angina despite medical therapy in 19 (coronary artery bypass surgery in 7 and coronary angioplasty in 12). By univariate analysis, the predictors of future cardiac events were a history of previous myocardial infarction (odds ratio 5.5, P < 0.02), multivessel coronary artery disease (odds ratio 9.6, P < 0.0002), angina during atrial pacing (odds ratio 5.1, P < 0.05), abnormal scintigraphy (odds ratio 17.1, P < 0.001) and reversible perfusion defect after pacing (odds ratio 7.9, P < 0.002). By multivariate analysis, multivessel disease (P < 0.004) and reversible perfusion defect after pacing (P < 0.02) were the only independent predictors of future cardiac events.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223734 TI - Digital radiofrequency echocardiography in the detection of myocardial contrast following intravenous administration of Albunex. AB - Conventional contrast echo techniques have proved inadequate for the detection of myocardial perfusion using intravenously injected echo contrast agents because of the limitations and relative insensitivity of standard echocardiographic equipment. In order to avoid these problems, we have obtained pure digital radiofrequency ultrasound data from the left ventricle and myocardium during i.v. contrast myocardial perfusion echo studies. In 30 patients, following coronary arteriography, i.v. injections of the echo contrast agent Albunex (sonicated human serum albumin) in two doses of 0.08 and 0.22 ml.kg-1 were administered during digital radiofrequency echocardiography sampling of data from the myocardium and left ventricular cavity. Analysis of mean integrated backscatter (MIB, a measure of the total ultrasound energy) was performed before, during and after Albunex injection. The data were also analysed for a shift in frequency spectrum which could be caused by resonance of the Albunex contrast microspheres in the heart, a phenomenon which has been previously demonstrated in vitro. Digital radiofrequency ultrasound data were successfully obtained and analysed in 23 patients. In 31 segments where reasonable resting perfusion was expected, there was a significant increase in MIB from 0.644 to 1.245, P < 0.001 and time intensity curves could be constructed showing wash-in and wash-out of contrast from the myocardium. In 10 segments supplied by significantly diseased vessels, MIB intensity increased from 1.044 to 1.874, P = 0.054. In myocardial segments supplied by non-diseased vessels, microsphere resonance also caused a drop in mean frequency of 140 KHz, P < 0.001, and permitted similar temporal analysis of myocardial perfusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223735 TI - Determination of agreement between cardiopulmonary and standard ECG stress testing in coronary artery disease. AB - The agreement between the results of standard ECG (CX) and cardiopulmonary (CPX) exercise stress tests performed in randomized sequence was evaluated in 40 patients with known coronary artery disease but who were not taking cardioactive therapy. Systolic blood pressure and heart rate were significantly higher during CPX only at low workload (less than 100 W). Exercise time and rate-pressure product at both peak exercise and ischaemic threshold were not significantly different between the two tests, even though their variability exceeded the value of 20%, which is generally accepted as the cut-off point for defining CX parameters as reproducible. However, the metabolic response to exercise, assessed by means of blood lactate kinetics analysis, was highly reproducible between the two tests. We conclude that the provocative role of exercise testing is not altered by the gas exchange analysis technique used in CAD patients. However, the common indexes of myocardial as well as of global physical performance may be influenced, thus requiring caution in comparing data with those derived from CX or from reference values. PMID- 8223736 TI - Enhanced sensitivity for detection of coronary artery disease by addition of atropine to dipyridamole echocardiography. AB - Dipyridamole echocardiography test (DET) has gained acceptance due to its safety, feasibility, diagnostic accuracy and prognostic power. The main limitation of the test is a less than ideal sensitivity in some patient subsets, such as those with limited coronary artery disease. Atropine with dipyridamole might theoretically combine to become a synergistic ischaemic stress test, by increasing myocardial oxygen demand through chronotropic stress and by reducing flow supply through a shortening of the diastolic interval under maximal coronary vasodilation. The aim of this study was to assess the effects of the addition of atropine to DET. Three hundred and twenty-one patients (age = 58 +/- 9 years), referred for testing in the echo lab, were initially studied by DET. Of these, 151 were stopped during or within the 2 min following dipyridamole infusion because of achievement of a predetermined end-point: obvious echocardiographic positivity (n = 137), severe chest pain (n = 3), diagnostic ST segment changes (n = 7) or limited side effects (n = 4). In another three cases, atropine was not given due to a history of glaucoma or severe prostatic hypertrophy. In the remaining 167 patients with a negative DET test, atropine (0.25 mg intravenously, repeated every min up to a maximum of 1 mg, if necessary) was added, starting 3 min after the end of the dipyridamole infusion. The dipyridamole-atropine echo test (DETA) was positive in 32 and negative in 135 patients, and no major side effects occurred in any patient.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223737 TI - Diagnosis of heart tumours by transoesophageal echocardiography: a multicentre study in 154 patients. European Cooperative Study Group. AB - In a retrospective multicentre study, the diagnostic potential of transoesophageal 2D-echocardiography (TEE) as compared to precordial 2D echocardiography (TTE) was determined in 154 patients with primary or secondary tumours of the heart. Additionally, the value of standard diagnostic parameters, such as symptoms, X-ray of the chest and electrocardiogram were evaluated. In 84 patients (24 male, 60 female; age 20-85, mean 56.6 years) intracardial tumours were present, and 70 patients (37 male, 33 female; age 18-79, mean 44.3 years) presented with peri- or paracardial tumours. The main symptoms of patients with intracardial tumours were dyspnoea (60.7%), vena cava syndrome (22.2%) and chest pain (20.2%). Embolization was found in 11.9%. Left or right atrial enlargement was observed on chest X-ray in 23 patients, and echocardiographic abnormalities in 17 cases. The patients with peri- or paracardial tumours presented with dyspnoea in 51.4% of cases, loss in body weight in 20.0% and with vena cava syndrome and chest pain in 17.1%. The chest X-ray was abnormal in 56 patients. Unspecific ST segment changes in the electrocardiogram were observed in five, and arrhythmias in seven cases. Diagnosis of atrial myxomas was achieved by TTE in 95.2%, by TEE in 100%, by angiography in 78.4%, by computed tomography (CT) or magnetic resonance tomography (NMR) in 70%. Identification of the attachment point was made by angiography in 8.1%, by TTE in 64.5% and by TEE in 95.2%. In 22 patients with intracardial tumours (myxomas excepted) diagnosis was achieved by TTE in 90.9%, by TEE in 100%, by CT or NMR in 88.9% and by angiography in 50%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223738 TI - Postoperative and long-term prognosis of myotomy-myomectomy for obstructive hypertrophic cardiomyopathy: influence of associated mitral valve replacement. AB - Several surgical techniques have been proposed for obstructive hypertrophic cardiomyopathy (OHCM): myotomy, mitral valve replacement (MVR), or myotomy myomectomy (MM). We reviewed our series of 47 patients who had undergone surgery in order to determine their prognosis and to know whether MVR+MM was better than MM only. Left intraventricular gradient decreased from 86 +/- 34 mmHg to 15 +/- 20 mmHg (P < 0.0001). Postoperatively, three patients died from low cardiac output (6.4%); five died later. Annual mortality (including postoperative deaths) was 3.0%. Follow-up was 5.7 +/- 0.7 years. Survival was 87 +/- 11% at 12 years. After operation, 91% had NYHA class I or II dyspnoea (before surgery this had been 28%); chest pain was CCS class I in 88% (vs 47%); 12% had had syncope since operation (vs 53%). The gradient decrease was larger in the MM+MVR group (P < 0.05). Survival and functional improvement were similar in the two groups. Mitral regurgitation decreased from 2.7 to 0 in the MM+MVR group (P < 0.0001), whereas it decreased from 1.5 to 1.2 in the MM group (ns). MM appears to be the procedure of choice. When mitral regurgitation is important or when an intrinsic disease of the mitral valve exists, the addition of MVR should be considered. PMID- 8223739 TI - Assessment of the influence of spinal cord stimulation on left ventricular function in patients with severe angina pectoris: an echocardiographic study. AB - Patients with severe, widespread coronary atherosclerosis and patients who have undergone several coronary artery bypass operations are often poor candidates for coronary bypass surgery (CABG). Spinal cord stimulation (SCS) has been shown to have an anti-anginal effect that is probably associated with an anti-ischaemic effect. In the present investigation, 15 patients with severe angina (mean age 64 years, range 49-71) were studied. All patients had a history of intractable angina pectoris despite optimal medical treatment and previous coronary bypass operation. The patients had multi-vessel disease and graft occlusion or graft stenosis on postoperative coronary angiograms. Left ventricular function was assessed echocardiographically at rest and during provocation with adenosine infusion in a control session without treatment and during treatment with SCS. The recovery time was at least 3 h. The decrease in the ejection fraction during adenosine infusion was more pronounced in the control situation (44 to 37%; P < 0.05) than during SCS (48 to 44%; ns), and the time to echocardiographic signs of dysfunction and to anginal pain during adenosine infusion was significantly prolonged during SCS (P < 0.001). In addition, the recovery time for these parameters was shorter during SCS (P < 0.001). It is concluded that the deterioration in left ventricular function during adenosine provocation was less pronounced with SCS than without. This possible anti-ischaemic effect is in agreement with results from earlier studies. PMID- 8223740 TI - Renal effects of low dose prazosin in patients with congestive heart failure. AB - Activation of the sympathetic nervous system may contribute to the renal vasoconstriction and sodium retention seen in congestive heart failure. Previous studies in congestive heart failure patients employing large doses of prazosin that lowered systemic blood pressure have been generally disappointing. The renal haemodynamic and segmental tubular effects of low non-depressor doses of prazosin (0.25 mg and 0.50 mg) were examined in eight female patients with mild to moderate congestive heart failure. Segmental tubular function was assessed by the lithium clearance method. Compared to placebo, prazosin caused a significant increase in urinary sodium excretion (from 56 +/- 7 to 92 +/- 7 mumol.min-1, P < 0.01), paralleled by significant increases in fractional excretion of sodium and lithium. Glomerular filtration rate and effective renal plasma flow were not altered by prazosin. Prazosin pre-treatment did not alter any of the renal responses to frusemide treatment (mean dose 85 +/- 14 mg). This study demonstrates that low non-depressor doses of prazosin have a clear natriuretic effect in congestive heart failure patients, which is predominantly established by interference with tubular reabsorption. PMID- 8223741 TI - Cardioprotection by nisoldipine: role of timing of administration. AB - Nisoldipine was administered at 10(-9) M, a dose lacking negative inotropism, to isolated and perfused rabbit hearts submitted to 60 min ischaemia (1 ml.min-1) followed by 30 min reperfusion. The drug was delivered either 30 min before ischaemia, at the onset and after 30 min of ischaemia and during reperfusion only. Cardiac protection was evaluated in terms of recovery of left ventricular pressure during reperfusion, release of creatine phosphokinase (CPK), mitochondrial function, tissue content of adenosine triphosphate (ATP) and creatine phosphate (CP), calcium homeostasis and the occurrence of oxidative stress, established measuring content and release of reduced and oxidized glutathione. The cytoprotective action of nisoldipine occurs in the absence of negative inotropism and is closely related to the time of administration. Optimal myocardial preservation is achieved when nisoldipine is given before or at the onset of ischaemia. Prophylactic administration of nisoldipine improved the recovery of the developed pressure from 15.9 +/- 1.0 (SE) mmHg to 47.8 +/- 1.9 mmHg, P < 0.01 and reduced the release of CPK from 830 +/- 29 to 229 +/- 27 mU.min-1 g-1 wet wt, P < 0.01. The accumulation of tissue and mitochondrial calcium was reduced from 58 +/- 11 and 49 +/- 9 to 14 +/- 6 and 10 +/- 4 mmol.kg 1 dry wt respectively, P < 0.01. This resulted in a significant (P < 0.01) preservation of all indices of mitochondrial function, allowing a higher recovery of ATP and CP after reperfusion (from 4.1 +/- 0.7 and 10.0 +/- 0.6 to 16.1 +/- 1.0 and 29.9 +/- 0.2 mumol.g-1 dry wt respectively, P < 0.001). Reperfusion induced myocardial accumulation and release of oxidized glutathione were reduced from 0.493 +/- 0.07 nmol.mg-1 protein and 0.768 +/- 0.063 nmol.min-1 g-1 wet wt to 0.225 +/- 0.07 and 0.157 +/- 0.038 respectively, P < 0.01. Similar data were obtained when nisoldipine was given at the time of ischaemia, while administration 30 min after the onset of ischaemia showed only a trend towards protection. Nisoldipine lost its protective effect when given on reperfusion. A multifactorial analysis of the data suggest that the cardioprotective effect of nisoldipine is related to the maintenance of membrane integrity, possibly since nisoldipine is highly lipophilic. PMID- 8223742 TI - Electrophysiology of the ageing rabbit and cat sinoatrial node--a comparative study. AB - Intrinsic properties of the human sinoatrial (SA) node have been shown to decline with age. In the present study we aimed at investigating the underlying mechanisms of age-dependent changes in intrinsic cycle length and sinoatrial conduction time. To this end, the cycle length and transmembrane potentials of the SA nodes of rabbits (2 days-5.6 years) and cats (6 weeks-18 years) were recorded and nodal conduction was reconstructed. The size of the SA nodes was measured in Sirius Red stained sections. Cycle length increases with age in both the rabbit and cat SA node, and in both species cycle length is dependent on diastolic depolarization rate and action potential duration. Nodal action potential duration increases with age in both rabbit and cat, whereas diastolic depolarization rate decreases in the cat only. The location of the primary pacemaker is not related to age. With age, sinoatrial conduction time increases in both species as a result of an enlargement of the area with low phase 0 upstroke velocities. The size of the SA node of adult animals does not increase with age. PMID- 8223743 TI - Ondine's curse and neurally mediated syncope--a new and important association. AB - A 6-year-old boy with congenital central hypoventilation syndrome (Ondine's curse) presented with presyncope. Investigation revealed a diagnosis of neurally mediated syncope. This previously unreported association provides strong evidence that autonomic dysfunction is implicated in both these conditions. PMID- 8223744 TI - A case of infective endocarditis in a farmer caused by Streptococcus equinus. AB - The following is a case report of a farmer with documented aortic valve disease who developed bacterial endocarditis secondary to Streptococcus equinus. This is a rare pathogen in man and its acquisition in this case may be related to the subject's occupation. PMID- 8223745 TI - Congenital aneurysm of the muscular interventricular septum. AB - Cardiac aneurysms are rare in the paediatric age group, and are classified as congenital when the aetiology cannot be demonstrated. We report a 1-year-old patient with a congenital aneurysm of the muscular interventricular septum. Ventricular septal aneurysmectomy was performed successfully. PMID- 8223746 TI - Pharmacological therapy of arrhythmias complicating dilated cardiomyopathy- implications of the arrhythmogenic substrate. AB - Dilated cardiomyopathy is a global myocardial abnormality in which it is likely that there are relatively homogeneous electrical conditions. The pattern of arrhythmic complications, especially atrial ectopic beats, ventricular ectopic beats and the brief salvoes of polymorphic ventricular tachycardia which are characteristic of the condition, suggest that triggered automaticity is a more likely arrhythmia mechanism than reentry. Although treatment with 'conventional' antiarrhythmic agents has an important place, drugs which alter myocardial loading conditions (and thus, thereby, a possible mechanism of triggered automaticity) may be effective. Therapeutic strategies directed against the basic disease processes are in an early stage of clinical development but they hold great promise for the future. PMID- 8223747 TI - Pharmacological therapy in coronary heart disease: prevention of life-threatening ventricular tachyarrhythmias and sudden cardiac death. AB - Life-threatening ventricular tachyarrhythmias are the main reason for sudden cardiac death in coronary heart disease. In the majority of survivors of cardiac arrest, malignant tachyarrhythmias generate from a structurally fixed arrhythmogenic substrate following myocardial infarction without evidence of acute ischaemia. Thrombolysis in acute myocardial infarction improves the electrical stability as elucidated by electrophysiological studies and the signal averaged surface ECG. In post-infarction patients, beta-blockers provide significantly beneficial effects on arrhythmic outcome, particularly in the presence of impaired left ventricular function, whereas calcium antagonists and vasodilators are of no affect or may worsen the prognosis. In survivors of myocardial infarction, the prophylactic use of class I antiarrhythmic agents, which are able to suppress frequent single or complex premature ventricular contractions, cause worsening of the prognosis due to their proarrhythmic properties. However, arrhythmia suppression by antiarrhythmic agents selects patients who are at very low risk for arrhythmic death. Pilot trials using class III antiarrhythmic agents suggest beneficial effect on the reduction of sudden death mortality. As regards secondary prevention of malignant tachyarrhythmias in survivors of ventricular tachycardia or fibrillation, there is controversy about the importance of Holter monitoring or invasive electrophysiological testing in the evaluation of drug efficacy. In patients with severely impaired left ventricular function, pharmacological treatment is of limited efficacy. Even in cases of significant suppression of spontaneous or inducible tachyarrhythmias documented by Holter recording or programmed ventricular stimulation, the arrhythmic outcome is considerably poorer, but it could be influenced by implantable defibrillators. Amiodarone, as a potential alternative to class I antiarrhythmic agents, particularly in patients refractory to conventional antiarrhythmic drugs, shows only limited effects on long-term outcome, which is in part caused by the toxicity of this agent. There is substantial need for new drugs without proarrhythmic properties and particularly for those that correct abnormalities of the automatic nervous system. PMID- 8223748 TI - Role of interventional therapy in patients with coronary heart disease and life threatening ventricular tachyarrhythmias. AB - This study includes 357 patients with coronary heart disease and life-threatening ventricular tachyarrhythmias. Thirty-three patients (10%) underwent direct current (DC) catheter ablation (group 1), 121 (34%) mapping-guided surgery (group 2) and 203 (56%) received an implantable cardioverter defibrillator (ICD) (group 3). The mean follow-up was 35 +/- 25 months in group 1, 41 +/- 37 months in group 2 and 22 +/- 22 months in group 3. During follow-up, 6 patients (18%) died in group 1, 45 (37%) in group 2 and 46 (23%) in group 3. Sudden death occurred in two patients (6%) in group 1, 10 patients (8%) in group 2 and eight patients (4%) in group 3. Non-fatal recurrences group were observed in 19 patients (57%) in group 1 and in 16 patients (14%) in group 2. ICD discharges occurred in 175/203 patients (86%) in group 3 with a mean number of 18 +/- 28 ICD shocks per patient. Our data show that catheter ablation has a low incidence of sudden death but a high incidence of non-fatal recurrences. Mapping-guided surgery yields low rates of sudden death or recurrence. Patients with an ICD have a low sudden death rate. A treatment algorithm is necessary to evaluate patients at high risk for sudden death and to choose the right patient for the right treatment. PMID- 8223749 TI - Patient selection for ICD treatment and peri-operative mortality. AB - In view of the reported high risks resulting from the implantation of a cardioverter (ICD) and our own experience, we retrospectively evaluated the patients who received an ICD at our institution, so as to predict factors that could lead to a high peri-operative risk. The patients were divided into three groups according to their diagnoses and the surgical procedure. Group I: 10 patients with non-ischaemic cardiomyopathy, age 53.9 +/- 14.0 years, ejection fraction (EF) 33.1 +/- 11.8; group II:13 patients with coronary artery disease (CAD), diffuse arteriosclerosis, age 60.0 +/- 6.8 years, EF 31.4 +/- 9.1%; group III:12 patients, age 61.9 +/- 9.4 years, EF 41.2 +/- 8.6, CAD, concomitant bypass grafting. In cases of cardiomyopathy, cardiac complications (30%) were reversible by adequate drug therapy. However, five patients in group II (38.5%) died from cardiac complications, mainly frequent arrhythmias and subsequent pump failure. In group III no peri-operative complications occurred. Therefore the high mortality (overall 13.2%) in our series was exclusively due to patients suffering from severe CAD and unsuitable for revascularization. Considering the high risk after ICD placement, primary heart transplantation might be the better procedure in younger patients with end-stage CAD. PMID- 8223750 TI - Pathophysiology of supraventricular tachycardia. AB - Supraventricular tachycardia is due to altered automaticity, triggered activity or most commonly, reentry. Atrial flutter is most frequently due to reentry with counterclockwise activation in the right atrium with the left atrium acting as a bystander. The small wavefronts demonstrated during atrial fibrillation are less random than was previously thought. In the study of atrioventricular (AV) nodal reentrant tachycardia, recent evidence suggests that multiple pathways involving intranodal and perinodal tissue are probably involved. In the Wolff-Parkinson White (WPW) syndrome, accessory pathways near the AV node have been demonstrated with decremental properties, suggesting conduction through accessory AV nodal tissue. The mechanism for atrial fibrillation in the WPW syndrome is poorly understood, but there is evidence that accessory pathways are branched and may themselves support micro-reentry. In Mahaim nodoventricular pathways, the atrium is not essential for reentry, unlike atriofascicular reentrant circuits. Atriofascicular pathways consist of AV node-like tissue and are usually located in the right atrial free wall. PMID- 8223751 TI - Cardiac receptor activation and arrhythmogenesis. AB - New evidence has been accumulated allowing a better understanding of the physiology and pathophysiology of receptors in the heart. Three major receptor systems influence electrophysiological characteristics of myocardial cells and are critical in the development and prevention of cardiac arrhythmias: the adrenergic, the muscarinic and the adenosine systems. Although it has long been recognized that beta adrenergic stimulation is arrhythmogenic, only recently have the mechanisms of this arrhythmogenic effect been clarified. In addition, the contribution to arrhythmogenesis of alpha receptor stimulation, which has been overlooked for many years, has been recognized as an important accompaniment to the beta component, especially during hypoxia-ischaemia. On the other hand, it has been demonstrated that although direct electrophysiological effects of acetylcholine on the ventricle remain controversial, the antagonism of sympathetic activation by cholinergic stimulation may be important in preventing arrhythmias induced by a high sympathetic tone in the presence of myocardial ischaemia. More recently, the importance of the adenosine system has been better appreciated. Experimental studies have shown that adenosine receptor activation inhibits the adenylyl cyclase system by activating the Gi regulatory proteins. Activation of this pathway inhibits the development of adrenergic-dependent triggered activity in isolated cells and these have also been confirmed in man. It is likely that this effect is specific against triggered rhythms induced by adrenergic activation. Even though further research is needed to clarify fully the interaction of these three systems at the subcellular level, the pharmacological modulation of these cardiac receptors appears as a rational approach for refining the treatment of arrhythmias. PMID- 8223752 TI - The signal-averaged ECG: time-domain analysis. AB - During the past decade, the high-resolution electrocardiogram as a non-invasive technique for the detection of ventricular late potentials has developed from an experimental method into a routinely applied non-invasive method for risk stratification of patients after myocardial infarction. Meanwhile, several approaches have been developed for the detection of ventricular late potentials including time-domain analysis, frequency-domain analysis and spectrotemporal mapping. Clinical applications are no longer limited to patients after myocardial infarction, but cover a wider spectrum of different cardiac diseases. This review focuses on some methodological aspects as well as on the results and current clinical applications of the analysis of the signal-averaged ECG in the time domain. PMID- 8223753 TI - Beat-to-beat variability of ventricular late potentials in the unaveraged high resolution electrocardiogram--effects of antiarrhythmic drugs. AB - The aim of this study was to assess variability of ventricular late potentials (VLP) in patients with and without inducible ventricular tachycardia (VT), and the effects of antiarrhythmic drugs on VLP variability in the high-resolution electrocardiogram (HRECG). In 27 patients 90 s of unaveraged HRECGs were analysed before and 2 h after oral administration of 200 mg disopyramide, 400 mg mexiletine, 300 mg propafenone and 160 mg DL-sotalol. The duration of the QRS (QRSD) and the duration of the terminal low amplitude signal (LASD) was measured from each beat. Beat-to-beat variability was defined as standard deviation of the differences between consecutive beats. Patients with inducible sustained VT (n = 9) showed higher LASD variability than patients without inducible VT (12.3 vs 9.3 ms.beat-1, P < 0.01). Patients with VLP (n = 17), as defined by the signal averaged ECG, also had higher QRSD and LASD variability (11.8 vs 9.5 ms.beat-1, P < 0.05; 11.5 vs 8.2 ms.beat-1, P < 0.01, respectively) compared to those without VLP. All class I drugs lengthened QRSD and LASD in terms of the absolute values, but only propafenone increased QRSD and LASD variability (9.7 to 12.0 ms.beat-1, P < 0.01; 8.9 to 11.9 ms.beat-1, P < 0.01, respectively). In patients with inducible VT, sotalol decrease LASD variability from 14.3 to 9.3 ms.beat-1 (P < 0.05). We conclude that beat-to-beat VLP variability is increased in patients at a high risk of malignant arrhythmias.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223754 TI - Value of different non-invasive methods for the recognition of arrhythmogenic complications in high-risk patients with sustained ventricular tachycardia during programmed ventricular stimulation. AB - It is well known that patients with coronary heart disease and ventricular tachycardia show a high incidence of arrhythmogenic complications and sudden cardiac death. The best predictor of spontaneous ventricular tachycardia and sudden death in these patients is programmed ventricular stimulation, but this invasive method is limited to specialized institutions. The purpose of our study was to assess the predictive value of Holter monitoring, late potentials and heart rate variability as markers for these high-risk patients. We investigated 20 patients (18 m, 2 f, age range 31-79 years) with coronary artery disease documented angiographically (6 patients with single vessel disease (vd), nine patients with 2 vd, five patients with 3 vd) and previous myocardial infarction. Each patient underwent 24-h ambulatory monitoring with analysis of rhythm of heart rate variability (24-h spectral and non-spectral analysis) and a signal averaged ECG with late potential measurement. In all patients, sustained ventricular tachycardia was inducible during programmed ventricular stimulation. Late potentials were recorded in 12 out of the 20 patients (60%). Ventricular arrhythmias of Lown classes IVa, IVb or V were recorded in 12 patients. Analysis of heart rate variability compared to 20 age- and sex-matched healthy controls revealed a loss of parasympathic activity and increased sympathic activity in 16 of the 20 patients (80%). CONCLUSION: in this study, heart rate variability was the most sensitive method with which to recognize patients at a high risk of arrhythmogenic complications. Evaluation of the above parameters in large controlled clinical trials may help predict arrhythmogenic complications and sudden cardiac death. PMID- 8223755 TI - Magnetocardiography: supraventricular arrhythmias and preexcitation syndromes. AB - Magnetocardiography is an elegant non-invasive method with which to study the electrical activity of the heart. The localization of cardiac electric sources, such as arrhythmia foci, would, in particular, be an interesting clinical application. In addition the detection of different abnormalities that could lead to cardiac depolarization and repolarization gives insight into how cardiac arrhythmias and arrhythmia mechanisms are generated. We have studied patients with supraventricular arrhythmias, especially those suffering from the Wolff Parkinson-White (WPW) syndrome. Magnetocardiographic localization of the preexcitation site was performed in 26 WPW patients, with an average accuracy of 2 +/- 1 cm in comparison to the results obtained by invasive catheter mapping. By inspection of spatial isofield maps during atrial depolarization, the risk of atrial fibrillation was classified correctly in 11/20 (55%) patients with documented atrial fibrillation, and in 4/6 (67%) patients without atrial fibrillation. In a case of focal atrial tachycardia, magnetocardiographic localization of the origin of the arrhythmia was performed during tachycardia. In addition, the level of conduction block was successfully determined in six patients with third-degree congenital atrioventricular block. PMID- 8223756 TI - Magnetocardiography: ventricular arrhythmias. AB - Since 1985, magnetocardiographic (MCG) mapping has been carried out in patients with severe ventricular arrhythmias, related to primary cardiomyopathy and ischaemic heart disease, for the purpose of non-invasive localization of the arrhythmogenic substrate or to screen patients at risk of sudden death. Data reported by several authors have demonstrated that magnetocardiography is useful to investigate non-invasively some of the possible electrogenetic abnormalities underlying clinical arrhythmias and to identify patients at risk of sudden death through the detection of late magnetic fields and quantitative estimation of repolarization/depolarization abnormalities. In this paper all the above mentioned applications have been briefly reviewed. Experience based on 35 patients, with Lown class 3 or higher ventricular arrhythmias, who have been studied magnetically, to localize the site of origin of their ventricular arrhythmia and to calculate their Relative Smoothness Index is described. Reproducible MCG three-dimensional localization of both ventricular extrasystoles and sustained tachycardia proved possible, using even single channel instrumentation, with an accuracy in the order of 10 mm as demonstrated by inverse localization of a biomagnetic catheter. The RS index was significantly lower in patients with cardiomyopathy or sudden death, but this parameter needs further investigation, as methodological drawbacks could impair its clinical reliability. In conclusion, MCG localization of the arrhythmogenic substrate can be useful to guide aimed invasive electrophysiology, biopsy and ablation close to the arrhythmogenic substrate. PMID- 8223757 TI - Magnetocardiography: three-dimensional localization of the origin of ventricular late fields in the signal averaged magnetocardiogram in patients with ventricular late potentials. AB - The purpose of this study was to detect ventricular late fields recorded by a biomagnetic multichannel system in patients with ventricular late potential and to determine the site of these ventricular late fields non-invasively in three dimensions. Biomagnetic signals of sinus beats during a 5-min acquisition period simultaneously recorded by a 37-channel system Krenikon were averaged in all channels. Ventricular late fields were determined in each channel according to the algorithm of Simson for ECG data. For the localization process, baseline correction from the averaged non-filtered signals was performed at the end of the QRS complex under visual control. The single current dipole model within the homogeneous half-space was applied. Eight patients post myocardial infarction with ventricular late potentials (four with recurrent sustained ventricular tachycardia) and four healthy individuals were examined. In the normal subjects, no ventricular late fields were detected. However, ventricular late fields were found in all patients, and were localized in six patients within the border zone of myocardial infarction. In the four patients with ventricular tachycardia, a spatial coincidence of the site of origin of ventricular late fields and the site of origin of ventricular tachycardia determined by catheter mapping was found in two. It is concluded that magnetocardiography is able to detect ventricular late fields and can be used to determine their site of origin. PMID- 8223758 TI - Abnormal ventricular contraction patterns in patients with arrhythmogenic substrates using three-dimensional phase analysis. AB - Arrhythmogenic substrate diagnosis has been achieved by electrophysiological studies and best localized by successful radiofrequency ablation. Pre-invasive localization procedures have been based on surface ECGs and more recently on biomagnetism, but in addition to these electric and magnetic signals a mechanical signal may be utilized: the initial site of contraction may be detected by phase analysis during radionuclide ventriculography. Generation of three-dimensional data set of phases is achieved by incorporating the new emission tomography technique. The performance of this modified phase analysis has been investigated for the detection of the normal contraction pattern during sinus rhythm, the arrhythmogenic substrate of the WPW syndrome and ventricular tachycardia, and further, to define the limitations of the method in experimental studies on pigs. In 30 out of 44 patients with normal sinus rhythm and no ventricular lesion, a characteristic phase pattern was found. Physiologically, the initial site of contraction appeared to be paraseptal and in the anterior wall of the right ventricle close to the apex. In 13 patients with WPW syndrome and in seven with ventricular tachycardia, the phase data were compared to the electrophysiological study. In 14 of 20 there was a complete match, in 3 of 20 a mechanical focus was found in the area adjacent to the electric focus. From experimental pig studies with simulated stimulation, a spatial precision of at least 20 mm was found at a pre-excitation of 20 ms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223759 TI - The arrhythmogenic substrate of the long QT syndrome: genetic basis, pathology, and pathophysiologic mechanisms. AB - The Long QT syndrome (LQTS) is a relatively rare disorder. It has a major clinical impact as affected individuals are prone to syncope and sudden arrhythmogenic cardiac death. The LQTS comprises three groups of patients. The Jervell-Lange-Nielsen syndrome is characterized by an autosomal recessive pattern of inheritance and congenital neural deafness. The Romano-Ward syndrome shows an autosomal dominant pattern of inheritance and normal hearing. Patients with the sporadic form of LQTS have no evidence of familial transmission and have normal hearing. Imbalance of sympathetic cardiac innervation with predominance of the left stellate ganglion and an intrinsic myocardial defect leading to early afterdepolarization are the two pathogenetic mechanisms of LQTS discussed today. More recently a genetic basis for the Romano-Ward LQTS has been reported. The genetic linkage to the Harvey ras-1 gene provides the basis for a new hypothesis that an impairment of guanine nucleotide binding proteins is responsible for symptoms observed in LQTS. This paper discusses the genetic basis, pathology and pathophysiology of LQTS and tries to unify the different theories. PMID- 8223760 TI - Late potentials in arrhythmogenic right ventricular dysplasia. Prevalence, diagnostic and prognostic values. AB - A time-domain signal-averaged ECG was performed in 52 patients with arrhythmogenic right ventricular dysplasia (ARVD) proven by angiography, in 45 control subjects with normal hearts, and in 132 patients with various types of ventricular arrhythmias and no evidence of cardiac disease during routine cardiac examination. Based on the results of control subjects, patients with at least 2/3 abnormal parameters (QRS > or = 113 ms, LAS > or = 38 ms, RMS 40 < or = 16 microV) were considered as having late potentials (LP). Prevalence of LP was 75% (39/52) in patients with ARVD, 19% (25/132) in patients with apparently idiopathic ventricular arrhythmias, and 4% (4/45) in controls (P < 0.01). In ARVD, no relationship was found between LP and age, type of ventricular arrhythmia (sustained or not), or extent of the disease on angiography. A significant correlation was found between the values of two parameters (QRS and LAS length) and the delay between the first ventricular arrhythmia and the examination (r = 0.39 and 0.42 respectively, P < 0.01). Further examinations (echo and/or angiography) revealed underlying heart disease in 26/132 patients with apparently normal hearts, including 13 with ARVD. LP were present in 21/26 patients (81%) with diseased heart vs 4/106 (4%) in those with normal hearts after complete examination, giving a sensitivity of 86%, and a specificity of 96% for detecting underlying heart disease. During a follow-up of 3.1 +/- 1.2 years after signal-averaged ECG, only one patient died from heart failure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223761 TI - The arrhythmogenic substrate--diagnostic and therapeutic implications: hypertrophic cardiomyopathy. AB - Hypertropic cardiomyopathy provides an important model for the study of mechanisms of sudden death. Initiating factors for sudden cardiac death may include sinus tachycardia and paroxysmal supraventricular arrhythmias. The haemodynamic effects of such triggers may be modulated depending on the patients exercise haemodynamics, peripheral vasomotor control and autonomic function. Myocardial disarray provides the underlying histological substrate for the lethal ventricular arrhythmias that represent the final common pathway. Attempts at individual risk stratification for sudden cardiac death using conventional risk factors and non-invasive techniques such as Holter monitoring identify a large group of patients, many of whom will not experience sudden cardiac death. Conventional electrophysiological methods such as programmed ventricular stimulation do not add to risk stratification inducing a variety of non-sustained or non-specific ventricular arrhythmias. A novel electrophysiological technique described in this article offers more promise in this regard. PMID- 8223762 TI - Cellular electrophysiological properties in myocardial infarction. AB - Important ventricular arrhythmias consequent upon myocardial infarction have several pathophysiological features. Because the arrhythmogenic substrate following myocardial infarction gradually changes different electrophysiological and biochemical determinants can be related to ventricular arrhythmias at distinct periods. In acute ischaemia and infarction, multiple ionic and metabolic changes result in marked electrophysiological inhomogeneity at the cellular level, in which post-repolarization refractoriness and cellular uncoupling are involved in conduction disturbances. Twenty-four to 72 h after coronary occlusion (subacute phase), action potential abnormalities, in addition to abnormal impulse generation in surviving cell layers within and in the border zone of the infarcted area, contribute mainly to arrhythmias. In healed infarcted myocardium, changes in intercellular impulse propagation as well as non-uniform anisotropic cardiac tissue play a major role in the maintenance of arrhythmias, whereas the initiating mechanisms are less well defined. PMID- 8223763 TI - Supraventricular tachycardia and pre-excitation syndromes: pharmacological therapy. AB - Tachyarrhythmias which originate above the bifurcation of the bundle of His or incorporate tissue proximal to it are classified as supraventricular tachyarrhythmias (SVT). Primary treatment of SVT attempts to influence the underlying disease. Therapy is subdivided into drug therapy, electrotherapeutic tools (e.g. antitachycardia pacemakers, catheter ablation) and antiarrhythmic surgery. Antiarrhythmic agents which slow conduction and suppress premature beats are efficient for emergency and long-term treatment of supraventricular tachycardias. We evaluated some of the most relevant antiarrhythmic drugs for SVT including propafenone, diprafenone, cibenzoline, lorcainide and sotalol; in addition, usage and efficacy of quinidine/verapamil, disopyramide, amiodarone, ajmaline, adenosine and flecainide are summarized. The principles for acute management of tachycardia episodes with narrow and broad complexes are outlined. The reason for the selection as well as the efficacy in the termination of the tachycardias is described for different antiarrhythmic agents including verapamil, adenosine, ajmaline, propafenone and flecainide. PMID- 8223764 TI - Surgical treatment of the Wolff-Parkinson-White syndrome--experiences in 120 patients. AB - Since January 1984, 120 patients (78 male, 42 female; aged 3 to 66 years) with the Wolff-Parkinson-White (WPW) syndrome have been operated upon. The indication for surgery was documented recurrent, paroxysmal tachycardia refractory to medical treatment in 118 cases. Twenty-two patients (18%) had additional heart disease. One hundred and twenty patients had a total of 140 accessory pathways (AP). AP were localized at the left free wall in 63% (87 AP), at the right free wall in 24% (35 AP), and septally in 13% (18 AP). Sixteen patients (13%) had multiple AP (12 patients had two and four patients had three AP), 124 AP were known pre-operatively (88%), 133 were localized intra-operatively (94%) and seven were diagnosed during re-operation (6%). Surgery in 28 patients with left lateral AP was via the epicardial approach and the endocardial approach in 59. Of 15 patients with right lateral AP, the surgical approach was epicardial in two and transmural in 13. A cryosurgery was also used in 117 patients. Fifteen patients suffered recurrences, in 12 of whom repeat surgery was required. One hundred and thirty-six AP (97%) were dissected successfully, of which 14 (10%) were ablated during re-operation. All patients survived the initial operation, but two patients died after re-operation. One patient is pacer-dependent due to persisting postoperative atrioventricular (AV) block. We conclude that surgical dissection of accessory pathways can be offered as an alternative to non-surgical treatment modes, at a low risk and with a high success rate. PMID- 8223765 TI - Recent advances in bone densitometry. PMID- 8223766 TI - First experience in healthy volunteers with technetium-99m L,L ethylenedicysteine, a new renal imaging agent. AB - Animal studies have indicated that technetium-99m L,L-ethylenedicysteine (99mTc L,L-EC) may be a promising tracer agent for renal function studies. We have performed a paired study with 99mTc-mercaptoacetyltriglycine (99mTc-MAG3) and 99mTc-L,L-EC in six male volunteers. In both cases, iodine-131-labelled o iodohippurate was co-injected as an internal biological standard. The analog images between 0 and 30 min p.i. were of identical diagnostic value for both tracer agents. The two renograms were similar in all volunteers. The mean 1-h plasma clearance for 99mTc-MAG3 and 99mTc-L,L-EC was significantly different, respectively 382.9 +/- 17.1 ml/min per 1.73 m2 versus 460.2 +/- 47.7 ml/min per 1.73 m2 (P < 0.003). The urinary excretion after 30 min p.i. was 69.4% +/- 5.6% of the injected dose for 99mTc-MAG3 versus 66.5% +/- 2.5% for 99mTc-L,L-EC (P > 0.05) and after 60 min p.i. respectively 83.1% +/- 3.9% versus 79.8% +/- 4.3% (P > 0.05). 99mTc-L,L-EC has a very low plasma protein binding (31% +/- 6.8%) as compared to 99mTc-MAG3 (88% +/- 5.2%) and a larger volume of distribution. Although the exact mechanism responsible for the high plasma clearance of 99mTc L,L-EC is not yet fully known, we conclude that this new agent merits further clinical evaluation in patients to establish its value as a renal radiopharmaceutical. PMID- 8223767 TI - Dosimetry of iodine-123 iodobenzamide in healthy volunteers. AB - The distribution of the dopamine D2-receptor specific ligand iodine-123 (S)-(-)-2 hydroxy-3-iodo-6-methoxy-N[(1-ethyl-2- pyrrolidinyl)methyl]-benzamide (123I-IBZM) was investigated in human adults from whole-body scans, blood samples and urine collected up to 48 h after injection. Results from the present study performed in six healthy volunteers were combined with those of five volunteers from a previous study. Using the brain, liver, lungs and spleen as source organs, the MIRD method was applied to calculate the absorbed radiation dose of the radioligand in various organs. The thyroid (despite blockage), gallbladder wall, large intestinal walls and spleen received the highest absorbed doses. The average effective dose equivalent of 123I-IBZM for adults was estimated to be 0.034 mSv/MBq. The absorbed dose to the thyroid may be a limiting factor for 123I IBZM studies in children. PMID- 8223768 TI - The influence of image resolution on the positron emission tomographic measurement of caudate glucose consumption. AB - The aim of this study was to investigate the influence of image resolution on (a) relative and absolute values of caudate glucose consumption (rCMRGlc) determined by positron emission tomography (PET), and (b) the detection of significant differences in these metabolic values between groups of subjects. For this purpose, raw data of cerebral accumulation of fluorine-18 fluorodeoxyglucose (FDG) obtained in 11 normal subjects and in nine patients with unilateral thalamic infarction were reconstructed using filtered backprojection with four different cut-off frequencies (CFs), yielding images with a transaxial resolution of 5.7, 7.1, 8.9 and 11 mm (full-width at half-maximum; FWHM). Absolute values of caudate rCMRGlc decreased significantly by more than 30% over the range of image resolutions studied. Bilateral ratios of caudate rCMRGlc were insensitive to variations in image resolution. Levels of significance assessing the differences in mean metabolic values between patients and controls were all below 0.01. They were, however, slightly better at image resolutions of 7.1 and 8.9 mm than at a resolution of 5.7 mm. These data indicate (a) that relative values of rCMRGlc are better suited to compare quantitative results from different PET cameras than are absolute values, and (b) that the CF used for the filtered back-projection exerts a small but not negligible influence on levels of significance assessing differences in metabolic values between groups of subjects. PMID- 8223769 TI - Repeated fluorodeoxyglucose positron emission tomography of the brain in infants with suspected hypoxic-ischaemic brain injury. AB - Positron emission tomography (PET) permits the study of cerebral metabolism in vivo. We performed repeated PET studies with fluorine-18 fluorodeoxyglucose (FDG) as a tracer to measure cerebral glucose metabolism for estimation of neurological prognosis in infants with suspected hypoxic-ischaemic brain injury. Fourteen infants (gestational age 35.3 +/- 4.67 weeks) were examined during the neonatal period (at age 38.4 +/- 2.7 weeks) and again at the age of 3.5 +/- 0.7 months; one further infant was studied only once at the age of 2.5 months. All children also underwent ultrasound examinations. Electroencephalography and computed tomography or magnetic resonance imaging were performed according to their clinical condition and their neurological development has been followed. FDG accumulated most actively in the subcortical areas (thalami, brainstem and cerebellum) and the sensorimotor cortex during the neonatal period. The repeated PET study showed that the uptake of FDG was markedly high and increased in all brain sections of infants with normal development (n = 11), whereas those with delayed development (n = 4) had significantly lower values (P < or = 0.005). PMID- 8223770 TI - Technetium-99m hexamethylpropylene amine oxime labelled leucocyte scintigraphy in ulcerative colitis and Crohn's disease. AB - Technetium-99m hexamethylpropylene amine oxime labelled leucocyte scintigraphy (LS) was performed on 45 occasions in 30 patients with ulcerative colitis and on 53 occasions in 34 patients with Crohn's disease. Serial images were taken following re-injection of the labelled leucocytes. The segmental extent of the inflammation and the grade of the leucocyte uptake were calculated, and compared with the laboratory results and colonoscopy findings. The sensitivity and specificity of LS proved higher in ulcerative colitis (87% and 93%) than in Crohn's disease (53% and 89% in cases with large intestine involvement, and 82% and 100% in cases with small intestine involvement). The activity of the process determined by LS correlates with the alpha 2-globulin level (r = 0.47), fibrinogen level (r = 0.50), fS iron level (r = -0.57), sedimentation (r = 0.44), leucocyte count (r = 0.38), platelet count (r = 0.34) and Best index (r = 0.31) in ulcerative colitis, but not in Crohn's disease. PMID- 8223771 TI - Cholescintigraphy in the diagnosis and assessment of benign papillary stenosis. AB - Benign papillary stenosis (BPS) is an uncommon condition, the diagnosis of which is often difficult. In this report cholescintigraphy with technetium-99m diethyl acetanilido-iminodiacetic acid has been evaluated for the diagnosis of BPS, as well as for assessing the effect of treatment by endoscopic sphincterotomy (ES). In 12 patients with BPS, cholescintigraphy was performed before and after ES and the findings compared with those in ten controls. Time-activity curves from regions over the liver and bowel were generated. A significant difference was found in respect of the time for maximum activity (Tmax) over the liver before and after ES. The time for occurrence of bowel activity (Texcr) also showed significant differences before and after ES. Considering both Tmax and Texcr mean values plus two standard deviations in the control group, all BPS patients would have been detected and there would have been no false-positives among the controls. It is concluded that cholescintigraphy should be used in the diagnosis of BPS. The method has high sensitivity and should be applied as an early diagnostic procedure to exclude BPS in patients with clinical suspicion of the disorder. The method can also be used for assessment of endoscopic treatment (ES). PMID- 8223772 TI - Noradrenaline depletion in patients with coronary artery disease before and after percutaneous transluminal coronary angioplasty with iodine-123 metaiodobenzylguanidine and single-photon emission tomography. AB - Iodine-123 metaiodobenzylguanidine (MIBG) is a noradrenaline analogue which can be used as a tracer to investigate the cardiac sympathetic nervous system. Regional ischaemia leads to noradrenaline depletion with functional denervation which can be demonstrated by reduced MIBG uptake. In order to evaluate the reversibility of ischaemia-associated damage to the sympathetic nervous system, neuronal scintigraphy with 123I-MIBG and myocardial rest and stress perfusion scintigraphy with technetium-99m sestamibi was performed in 16 patients with coronary artery disease before and 3-4 months after percutaneous transluminal coronary angioplasty (PTCA). Partial re-innervation occurred in five patients, the degree of stenosis of remaining lesions being estimated by repeat angiography to be below 40%. Unchanged MIBG defects could be confirmed in four patients with residual lesions of between 40% and 50%. Increased MIBG defects were shown in three patients with significant restenoses of more than 70%. In all patients the neuronal defects exceeded the ischaemia-induced or scar-associated perfusion defects. Three patients dropped out of this study: one for technical reasons, one due to emergency aortocoronary bypass surgery and one due to diabetic polyneuropathy. This investigation shows that the sympathetic nervous system is highly sensitive to ischaemia. Further studies need to be done to assess the conditions allowing re-innervation after PTCA. PMID- 8223773 TI - Initial experience with single-photon emission tomography using iodine-123 labelled 2 beta-carbomethoxy-3 beta-(4-iodophenyl) tropane in human brain. AB - The iodinated cocaine analogue 2 beta-carbomethoxy-3 beta-(4-iodophenyl)tropane ([123I]beta-CIT), a new dopamine transporter, was preliminary tested in human brain. Two normal volunteers and two patients with Parkinson's disease were imaged with a high-resolution single-photon emission tomography scanner. The specific binding of [123I]beta-CIT in the basal ganglia and thalamus was high in normal volunteers. In addition, there was relatively intense uptake in the medial prefrontal area. Patients with Parkinson's disease who were older than controls showed significantly lower specific binding in the basal ganglia and thalamus and no uptake in the medial prefrontal cortex. This decrease in the dopamine transporter may be age related. PMID- 8223774 TI - Where do we stand with the International Commission on Radiological Protection Report 60? AB - We have examined the implications of ICRP60 for nuclear medicine. Radiation doses to staff are currently low and reduction of the dose limits will have little impact. However, the proposed figures for the fetus may have implications for pregnant women where the workloads are high. With nursing staff on the ward, laboratory staff or indeed departmental porters there seems little problem. Radiation dose to the fingers is, however, a key factor, and ensuring that no further restrictions are placed on the proposed dose limits is important. The concept of constraints is becoming clearer but will need further thought, particularly with regard to exposure of the general public. The previous basis for declaring controlled and supervised areas has been abandoned, and the situation is now less clear. A most important area of debate will be the patient who is a controlled area, and great care will have to be taken to ensure that the number of patients in this category is not increased so that many more patients need to be kept in hospital. Exposure of the general public is also an issue and expanding the concept of medical exposure to include carers is an important development. This should prevent a number of unnecessary restrictions and adds weight to the point that two waiting rooms are not justified in nuclear medicine departments. ICRP recommendations will soon be incorporated into the legislation of member states and it is import for everyone to ensure that drafts are read carefully and that national proposals are not too restrictive. PMID- 8223776 TI - Avid thallium-201 uptake in hepatic carcinoid metastases. AB - While radiolabelled somatostatin analogues sensitively detect extrahepatic carcinoid tumour, intrahepatic metastases are frequently not visualised due to somatostatin accumulation in normal hepatic tissues. We report a case of avid thallium-201 uptake in multiple hepatic carcinoid metastases using single-photon emission tomography (SPET) and compare this to a SPET technetium-99m sulphur colloid scan in the same patient. The 99mTc sulphur colloid images demonstrate multiple focal defects at the site of known metastases in the hepatic right lobe (confirmed on both CT and surgery). However, there is avid uptake of thallium in the metastases on comparative SPET slices. 201Tl may be a useful agent for the detection and localisation of carcinoid tumour and in particular of intrahepatic carcinoid metastases. PMID- 8223775 TI - Myocardial viability: what do we need? AB - Coronary revascularization in patients with chronic coronary heart disease (CHD) or acute myocardial infarction (AMI) is mainly based on factors such as coronary anatomy, ventricular function, accompanying diseases and the patient's biological age. Rest- or exercise-induced ischaemia should be proven before a bypass operation or percutaneous transluminal coronary angioplasty. Although a significant amount of ischaemic but still viable myocardium is a necessary condition for successful revascularization, the detection of viable myocardium is of major importance in a rather small subset of patients. These are patients with hibernating (or a combination of hibernating and stunned) myocardium in whom the aforementioned parameters do not yield an unequivocal result. Thallium-201 myocardial scintigraphy with re-injection or rest-redistribution is an established, proven and cost-effective way of detecting viable myocardium. Other methods such as positron emission tomography with different tracers or technetium 99m sestamibi SPET are discussed and compared to thallium-201 SPET. In conclusion, the detection of ischaemic but still viable myocardium is of importance in only a rather small subset of patients with CHD. In this context thallium-201 myocardial SPET is and still remains the method of choice. PMID- 8223777 TI - The ontogeny of the gut mucosal immune system and the susceptibility to infections in infants of developing countries. AB - In this review we summarize data on the human gut mucosa associated lymphatic tissues as part of the common mucosal immune system. Its embryonal-fetal and post natal ontogeny becomes severely distorted and compromised by mal-/undernutrition which is so prevalent in developing countries. Pathogenetic interdependencies exist between maternal-fetal undernutrition, the ontogeny of the immune system, constant antigenic stimulation of the mucosal immune system post-natally, and the 14 million deaths annually from infections in children below the age of 5 years in developing countries. A detailed knowledge of these interdependencies is required for effective prevention and treatment in an attempt to reduce the high morbidity and mortality rates of children in developing countries. PMID- 8223778 TI - Nitric oxide and prostacyclin lower suprasystemic pulmonary hypertension after cardiopulmonary bypass. AB - In a 3-week-old male newborn persistent suprasystemic pulmonary hypertension developed after surgical valvulotomy for a critical aortic valve stenosis. Because of a residual transvalvular pressure gradient of 35 mmHg and postoperative left as well as right ventricular dysfunction, treatment with inhaled nitric oxide (NO) and intravenously infused prostacyclin (PGI2) was attempted. Low-dose inhaled NO and low dose PGI2 corrected severe pulmonary hypertension and led to an increase in cardiac output. Treatment with NO but not PGI2 was accompanied by a rise in PaO2 and systemic blood pressure. Interruption of NO administration led to a rapid increase in pulmonary arterial pressure to suprasystemic levels. With continued i.v. PGI2 and decreasing concentrations of NO, severe pulmonary hypertension resolved after a few days suggesting that a transient endothelial dysfunction was partially responsible for pulmonary vasoconstriction. NO inhalation appears to be an effective new tool in the treatment of severe pulmonary hypertension following cardiac surgery. PMID- 8223779 TI - Decreased beta 2-adrenoceptor density and decreased isoproterenol induced c-AMP increase in juvenile type I diabetes mellitus: an additional cause of severe hypoglycaemia in childhood diabetes? AB - Little is known about the receptor and post receptor mechanisms of sympathoadrenal signal transmission in type I diabetes mellitus. Therefore, we examined the maximum binding of granulocyte beta 2-adrenoceptors and the in vitro c-AMP accumulation in lymphocytes of 24 children and adolescents with diabetes mellitus and 14 similarly aged healthy subjects. The number of high affinity beta 2-adrenoceptors on granulocytes correlated significantly with unstimulated (r = 0.6, P < 0.004) and with isoproterenol stimulated c-AMP values in lymphocytes (r = 0.68, P < 0.0007) showing the proportional changes of beta 2-adrenoceptors and c-AMP in two different cells. The number of beta 2-adrenoceptors on granulocytes was significantly reduced in diabetic as compared to healthy children (median 1397, range 599-3405 vs. 2205, 825-3200 beta 2-adrenoceptors per granulocyte, P = 0.014). Moreover, the percentage in vitro stimulation of c-AMP by isoproterenol in lymphocytes was significantly reduced in diabetic children as compared to healthy individuals (120%, 39%-278% vs. 225%, 66%-500%, P = 0.012). These results indicate a decreased sympathoadrenergic signal transmission in peripheral blood cells as a model for the liver probably contributing to severe hypoglycaemia in diabetic children. PMID- 8223780 TI - Malignant thymoma in a patient with growth hormone deficiency during growth hormone therapy. AB - Malignant thymoma was found in an 8-year-old Japanese boy with growth hormone (GH) deficiency who had received GH therapy for 3 years and 5 months. There may be a possible relationship between the occurrence of malignant thymoma and GH therapy. PMID- 8223781 TI - Childhood tuberculosis at a Swiss university hospital: a 2-year study. AB - The incidence of tuberculosis (TB) presenting to the University Hospital, Geneva, Switzerland, was studied over 2 years. Clinical data on all children receiving antituberculous drug therapy was collected. Of the 43 identified cases, 42% had active disease (i.e. clinical evidence of organ involvement), the remaining 58% being asymptomatic. All 43 children originated from countries other than Switzerland. Symptomatic, drug induced hepatitis necessitated temporary interruption of therapy in 2 children (5%). Contact screening yielded three new cases of pulmonary TB, three previously diagnosed cases and seven instances of a positive medical history in relatives living in other countries. Of the 34 families in this study, contact screening could not be completed in 8. The results of this study indicate that children with TB are primarily associated with families originating from countries other than Switzerland, particularly those in which TB remains endemic. Tuberculin skin testing should therefore be targeted at this group. Contact tracing has also been shown to be beneficial. PMID- 8223782 TI - Demonstration of Borrelia burgdorferi infection in a child with Guillain-Barre syndrome. AB - A 4.5-year-old child suffered from rapidly evolving motor weakness with paresthesia and radicular pain in both legs, accompanied by an isolated elevation of protein levels in CSF. Isoelectric focusing revealed oligoclonal IgG produced in the cerebrospinal compartment only. The diagnosis of Guillain-Barre syndrome was confirmed by electrophysiological studies. High antibody titres against Borrelia burgdorferi in the serum and specific antibodies in the CSF were demonstrable by ELISA and immunoblotting. B. burgdorferi specific DNA fragments were demonstrated in urine and CSF by means of a nested polymerase chain reaction (PCR), thus providing the borrelial aetiology and indicating B. burgdorferi infection. Complete recovery was observed after treatment with high dose immunoglobulin and ceftriaxone and control urine specimens were PCR negative. PMID- 8223783 TI - Body proportions in individuals with Turner syndrome. The Dutch Growth Hormone Working Group. AB - The body proportions in 191 individuals with Turner syndrome (TS) were investigated. At 3 years of age the mean sitting height in TS was normal, thereafter trunk growth was impaired, resulting in a standard deviation score (SDS) of -2.4 in the adult. From 3 to 12 years of age the mean SDS of leg length increased from -2.7 to -3.6; and then fell to -2.5. At 3 years of age the ratio of sitting height to leg length was 3.2 standard deviations (SD) above the normal mean. Thereafter the ratio slowly approached the normal percentiles. It was +0.6 SD in 15- to 18-year-old women. Thereafter it increased to 1.7 for adults with TS. Knemometric measurements in 32 individuals with TS and 32 controls revealed that in TS the upper legs were relatively shorter than the lower legs. We conclude that children with TS, and to a lesser extent adults, have a disproportionately short stature with relatively short legs whereas body proportions are almost normal in adolescents. PMID- 8223784 TI - Failure of calcitriol treatment in a patient with malignant osteopetrosis. AB - In an attempt to stimulate bone resorption, a 10-week-old infant with malignant infantile osteopetrosis was treated with high doses of calcitriol, a potent bone resorption stimulatory agent, combined with a low calcium diet to prevent hypercalcaemia. Although calcitriol administration was initiated at this very young age, our patient did not show any clinical, radiological, or histological improvement. Despite reports of positive results of this treatment in the literature, our patient did not reveal any signs of bone resorption. She eventually died from the complications of osteopetrosis at the age of 6 months after 88 days of therapy. PMID- 8223786 TI - The apparent role of blood transfusions in the development of retinopathy of prematurity. AB - During a 30-month-period, 184 very low birth weight infants from two Liverpool neonatal intensive care units were screened for evidence of retinopathy of prematurity (ROP). Seventeen clinical variables previously considered relevant to the development of ROP, blood gas and blood pressure data over the first 7 days, and the maximum stage of ROP reached in either eye were recorded, together with the need for cryotherapy and current visual status. Ninety-two infants developed any stage of ROP and 15 required cryotherapy or became blind. Logistic regression showed that only gestational age and frequency of blood transfusion were independently associated both with the risk of occurrence of ROP and its severity. PMID- 8223785 TI - Clinical and biochemical consequences of copper-histidine therapy in Menkes disease. AB - Menkes disease (MD) is an X-linked recessively inherited neurodegenerative disorder of copper (Cu) metabolism leading to death in early childhood. Symptoms are attributed to deficient activity of Cu-dependent enzymes. Limited experience has been reported concerning clinical and biochemical consequences of parenteral treatment with copper-(histidine)2-complex (Cu-His) in MD. Cu-His was administered in a 13-week-old boy with MD by daily intramuscular injections. After 6 weeks of therapy, Cu and caeruloplasmin in serum and Cu in CSF were normalized. The excessive dopamine level in CSF was corrected after 3 months of treatment. After 6 weeks of Cu supplementation, complete reduction of epileptic discharges, improved muscular tone and increased motor activities were observed. Developmental regression stopped and was replaced by a slight progression. Death at the age of 19 months was caused by septicaemia due to a fulminant urinary tract infection; there was no evidence of chronic Cu toxicity. These findings suggest that Cu-His supplementation may be a promising palliative treatment in MD. PMID- 8223787 TI - Renal clearance of bilirubin conjugates in newborns of different gestational age. AB - The renal excretion of bilirubin conjugates was analysed in 22 newborns. Bilirubin monoconjugate was the only metabolite detectable in urine samples and its renal excretion correlated with the creatinine excretion rate (r = 0.91). The renal clearance of bilirubin monoconjugates in newborns ranged between 380 and 2160 ml/1.73 m2 per 24 h (median: 790). According to the present findings the renal function should be monitored in newborns and infants with conjugated hyperbilirubinaemia. PMID- 8223788 TI - Comparison of umbilical arterial versus umbilical venous blood pH correlated with arterio-venous glucose difference and cardiotocographic score. AB - The value of clinical parameters and umbilical arterial blood pH as indicator of prenatal hypoxia is disputed. In a prospective study of 86 vaginally delivered full-term infants, cardiotocographic (CTG) findings obtained 0-30 min and 30-60 min before birth were compared to pH values, O2 and CO2 partial pressures and glucose difference in umbilical arterial and venous blood. CTG findings were expressed as a score, higher values indicating fetal hypoxia. The venous but not the arterial blood pH was significantly related to the later (0-30 min) CTG score. The arterio-venous glucose difference was significantly related to both CTG scores. There was a significant statistical relationship between glucose difference and venous but not arterial blood pH. The later CTG score (0-30 min) also correlated significantly with O2 and CO2 partial pressures and base excess in the umbilical vein of all vaginally born infants. If CTG is accepted as an objective indicator of fetal hypoxia before birth, the arterio-venous glucose difference, and in the investigated range of pH-values, umbilical venous blood pH are more suitable than the arterial blood pH to ascertain the peripartal situation of the newborn. PMID- 8223789 TI - The relation between neurological trauma parameters and long-term outcome in children with closed head injury. AB - In a study of 54 children with closed head injury, the relation between neurological severity indicators and the occurrence of long-term sequelae was investigated. Patient data were recorded during the clinical phase and during a 2 year follow up period. A correlation analysis identified early predictors for the occurrence of long-term sequelae. In particular, the duration of coma and of post traumatic amnesia correlated strongly with the occurrence of neurological, behavioural and intellectual residual sequelae but not with the more subjective complaints. PMID- 8223790 TI - Dystrophinopathy in two young boys with exercise-induced cramps and myoglobinuria. AB - Two young boys were referred for evaluation of metabolic myopathy because of elevated serum levels of creatine kinase, cramps and pigmenturia. Immunohistochemical studies of dystrophin in muscle biopsies showed reduced intensity of the stain with a patchy and discontinuous pattern in most fibers. In both patients dystrophin was undetectable by immunoblotting. DNA analysis of the dystrophin gene was not informative in one patient; in the other it revealed an in-frame deletion comprising exons 3-6. These observations suggest that the two patients are affected with an unusual phenotype of Becker muscular dystrophy. Dystrophin analysis should be included in the evaluation of patients with childhood-onset of recurrent myoglobinuria. PMID- 8223791 TI - Vitamin K1 and K2 status and faecal flora in breast fed and formula fed 1-month old infants. AB - Faecal vitamin K1 (VK1, phylloquinone) and vitamin K2 (menaquinone, MK) concentrations were measured in 12 breast fed and 9 formula fed 1-month-old infants. Faecal concentrations of VK1 and MK-5 to -9 were significantly higher in the formula fed than in the breast fed infants. There was also a tendency for higher total faecal MK (4-10) concentrations in the formula fed [geometric mean (95% confidence intervals); 8995.0 (3872.6, 20,893.0) pmol/g of dry faeces] than in the breast fed infants [2937.7 (1285.3, 6714.3), P = 0.051]. The numbers of streptococci and Escherichia coli were 100 and 10 times higher, respectively, in the faeces of the formula fed than in those of the breast fed infants. Faecal concentrations of MK-6, -7 and -8, and MK-8 were correlated with the numbers of streptococci and E. coli in the faeces, respectively. Serum VK1 and MK concentrations were measured in 9 out of 12 breast fed and eight out of nine formula fed infants. The serum VK1 concentration was much higher in the formula fed infants [average (95% CI); 2.20 (1.48, 2.92) pmol/ml] than in the breast fed ones [0.30 (-0.10, 0.70), P = 0.000], but MKs were not detected in the sera of most of the formula fed infants. PMID- 8223792 TI - A placebo controlled trial of fluticasone propionate in asthmatic children. AB - Fluticasone propionate is a synthetic steroid for use by the inhaled route. It's high topical potency and low systemic bioavailability make it suitable for use in asthmatic children. A total of 258 children were randomised in a double-blind study to receive fluticasone propionate (50 micrograms bd) as the dry powder formulation inhaled via a Diskhaler inhaler, or matched placebo (with current therapy) for 4 weeks throughout which time diary cards were completed. During clinic visits lung function and adrenal function were measured. Fluticasone propionate produced a significantly greater increase in morning peak expiratory flow rate (PEFR) (adjusted mean difference over days 1-28, 17 l/min (95% CI; 10, 24); P < 0.001) and evening PEFR (adjusted mean difference over days 1-28, 16 l/min (95% CI; 9, 23); P < 0.001). In addition, diary card symptom scores, beta 2 agonist rescue and clinic lung function improved significantly on fluticasone propionate. There were few adverse events and basal plasma cortisol remained within the normal range. In conclusion fluticasone propionate at 50 micrograms bd is superior to placebo (current therapy) in the treatment of childhood asthma with no evidence of adverse effects. PMID- 8223793 TI - Helicobacter pylori positivity. PMID- 8223794 TI - Temporomandibular joint involvement in familial Mediterranean fever. PMID- 8223795 TI - The Australian NHMRC Twin Registry: a resource for paediatric research. PMID- 8223796 TI - A proposition for the diagnosis and treatment of gastro-oesophageal reflux disease in children: a report from a working group on gastro-oesophageal reflux disease. Working Group of the European Society of Paediatric Gastro-enterology and Nutrition (ESPGAN). AB - In this paper, a Working Group on Gastro-Oesophageal Reflux discusses recommendations for the first line diagnostic and therapeutic approach of gastro oesophageal reflux disease in infants and children. All members of the Working Group agreed that infants with uncomplicated gastro-oesophageal reflux can be safely treated before performing (expensive and often unnecessary) complementary investigations. However, the latter are mandatory if symptoms persist despite appropriate treatment. Oesophageal pH monitoring of long duration (18-24 h) is recommended as the investigation technique of choice in infants and children with atypical presentations of gastro-oesophageal reflux. Upper gastro-intestinal endoscopy in a specialised centre is the technique of choice in infants and children presenting with symptoms suggestive of peptic oesophagitis. Prokinetics, still a relatively new drug family, have already obtained a definitive place in the treatment of gastro-oesophageal reflux disease in infants and children, especially if "non-drug" treatment (positional therapy, dietary recommendations, etc.) was unsuccessful. It was the aim of the Working Group to help the paediatrician with this consensus statement and guide-lines to establish a standardised management of gastro-oesophageal reflux disease in infants and children. PMID- 8223797 TI - Intravenous nicardipine in hypertensive children. AB - Fourteen hypertensive patients hospitalized in a paediatric intensive care unit were studied to evaluate safety and hypotensive efficacy of intravenous nicardipine. Systolic and diastolic blood pressure significantly decreased 1 h after the beginning of the treatment (1 microgram/kg per minute). Mean decrease in systolic blood pressure during the first 24 h was between 9.9% and 13.4% of the initial value. Mean lowering of diastolic blood pressure was between 16.7% and 25.6%. Nicardipine did not significantly affect heart rate with dose of 1 microgram/kg per minute. No clinical side-effects were observed. Nicardipine could be a first line drug for the treatment of hypertension in paediatric intensive care units. PMID- 8223798 TI - Hypomelanosis of Ito and precocious puberty. AB - We describe two girls with hypomelanosis of Ito, one of whom was demonstrated to have diploid-triploid mixoploidy in skin fibroblasts. Both had precocious puberty which was probably gonadotrophin-independent. The association of precocious puberty and hypomelanosis of Ito has not been previously reported. We have speculated on the possible mechanism of gonadotrophin-independent precocious puberty. PMID- 8223799 TI - Reduction of bone density: an effect of gonadotropin releasing hormone analogue treatment in central precocious puberty. AB - Gonadal steroids drive the significant bone mineral increase that occurs at puberty. Oestrogen deprivation in women results in bone loss. We investigated bone mineralization by single photon absorptiometry in girls with central precocious puberty (n = 13, age 3.8-8.5 years) before and during 1 year of treatment with gonadotropin releasing hormone analogue (GnRH-a = long-acting D Trp6-GnRH, 60 micrograms i.m. every 28 days). Before GnRH-a therapy, bone mineral density (BMD) was significantly higher in patients than in ten control girls matched for chronological age (patients 0.575 +/- 0.097 g/cm2, controls 0.433 +/- 0.049 g/cm2, P < 0.001). Patient BMD was not significantly different from that of ten control girls matched according to patient bone, age (0.550 +/- 0.046 g/cm2, P = NS). During GnRH-a treatment, pituitary-gonadal axis was suppressed and patient BMD significantly decreased (6 months: -6.0%, P < 0.002 vs baseline; 12 months: -8.0%, P < 0.001 vs baseline). We conclude that in girls with precocious puberty the activation of gonadal steroid secretion induces an increase in bone mineralization and that oestrogen deprivation by GnRH-a treatment caused a significant decrease in BMD. PMID- 8223800 TI - Evaluation of growth hormone in thalassaemic boys with failed puberty: spontaneous versus provocative test. AB - Growth hormone (GH) secretion was determined by evaluating ultradian GH profiles for 12 h and GH responses to insulin stimulated hypoglycaemia (ITT) in 28 stunted boys with beta-thalassaemia major aged 15.2-17.4 years, who presented with pubertal failure (FP). Healthy non thalassaemia prepubertal boys (n = 10) aged 7.5-8.8 years, were studied as controls. All patients had normal responses to ITT with peak GH levels > or = 15 mU/l. Basal GH concentrations (mean +/- sem) (1.65 +/- 0.03 mU/l vs 2.58 +/- 0.27 mU/l; P < 0.05) and the stimulated GH responses (peak GH = 15.4 +/- 0.20 mU/l vs 21.08 +/- 0.78 mU/l; P < 0.001) were significantly lower in the patients with failed puberty than in the controls, indicating that the FP patients had diminished GH reserve and secretory capacity. Moreover, all the GH peak parameters including the maximum spontaneous concentrations (MX-GH) and the area under the GH curve (AUC) were significantly lower in the thalassaemic patients than in the controls (MX-GH = 5.2 +/- 0.21 mU/l vs 20.42 +/- 0.14 mU/l; P < 0.001; AUCb = 421.22 +/- 4.31 mU/l vs 712.20 +/- 3.42 mU/l; P < 0.001). These observations suggest that the thalassaemic patients had endogenous neurosecretory GH deficiency (GHND). Priming with sex steroid did not cause any improvement in the spontaneous or stimulated GH secretory patterns in thalassaemic patients. It was noteworthy that in neither the patients nor the control subjects, was there a significant correlation between the maximum stimulated and the MX-GH concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223801 TI - Surgical treatment of Crohn disease in children and adolescents; how conservative can the paediatrician be? AB - Thirty-eight children (21 male, 17 female, age 3-18 years), treated for Crohn disease in two Dutch university centres, were retrospectively studied in order to evaluate the results of conservative treatment and to find out in what way surgical treatment in this age group may have differed from treating adults with this disease. Both groups had an equal distribution of age and sex. Diarrhoea with discharge of blood and mucus, abdominal pain, nausea/vomiting, weight loss, fever and general discomfort were the most frequent presenting symptoms. Twenty three children (60%) showed signs of malabsorption; 4 children (10%) had growth retardation. In 27 children (70%), 63 surgical procedures were performed (2.4 operations per child). There was no surgical mortality. Most operations were performed for ileocolitis and colon-only localizations needed most re-operations. Of the surgical procedures performed, 55% were excisional procedures. Already 3 years after the onset of symptoms, 50% of all children had had their first resection, whereas in adults, 50% of the patients undergo surgery 8 years after disease onset. Eight children were treated with split ileostomy. In only one of these children, operated for non-toxic colitis and severe steroid-dependent growth retardation, could the colon eventually be saved. The time between the onset of symptoms and the first operation seems to be shorter in children compared to adults. Severe malabsorption and growth retardation are additional specific indications for surgery for Crohn disease in childhood. The latter combined with non-toxic colitis, may perhaps be the only indication left for performing split ileostomy in Crohn disease. PMID- 8223802 TI - Growth and growth hormone in children during and after therapy for acute lymphoblastic leukaemia. AB - Growth impairment and growth hormone (GH) deficiency have been reported in children treated for acute lymphoblastic leukaemia (ALL). We have studied growth and GH secretion in a group of 50 patients, affected by ALL, during a 2- to 5 year period after diagnosis, and in 12 "long-term-survivors". We observed a significant decrease in growth velocity during the 1st year (in particular during the first 6 months) of therapy and a catch-up growth after the end of therapy. "Long-term survivors" did not exhibit a significant reduction of height standard deviation score (SDS), as compared to height SDS at diagnosis. None of the patients showed GH deficiency. Our data indicate that chemotherapy significantly affects growth of patients treated for ALL, whereas radiotherapy-at the doses used in this study-does not induce GH deficiency, at least not within 9 years after diagnosis. PMID- 8223803 TI - Prepubertal melanoma in a medium-sized congenital naevus. AB - A case of malignant melanoma in a medium-sized congenital naevus in a prepubertal girl is presented. Risk factors for developing melanoma during childhood include giant congenital naevi, dysplastic naevus syndrome and xeroderma pigmentosum. The lifetime risk of melanoma associated with giant congenital naevi has been estimated to be 4%-20%; the risk associated with small and medium-sized congenital naevi however remains controversial. In the latter lesions, malignant transformation is considered an almost exclusively postpubertal phenomenon, in contrast to giant congenital naevi where it often occurs prior to puberty. In our patient, malignant transformation in a medium-sized congenital naevus occurred before puberty. We suggest that the true incidence of malignant transformation within these lesions and the time at which it occurs, should be documented by prospective studies and that not only the giant congenital naevi but also the smaller congenital naevi should be considered for prophylactic excision in early childhood. PMID- 8223804 TI - Relapse of herpes simplex encephalitis after acyclovir therapy. AB - A case of herpes simplex encephalitis in a previously healthy 17-month-old girl relapsing 1 week after completion of a 10-day acyclovir therapy is described. The child responded favourably to a second 10-day course with acyclovir. The time of relapse and the satisfactory response to a second course of acyclovir treatment indicate that the first course of therapy was inadequate for eradication of herpes simplex virus infection. PMID- 8223805 TI - Thrombocytopenia in a case of neonatal mumps infection: evidence for further clinical presentations. AB - We report a rare case of congenital mumps infection in a newborn girl. Her mother developed bilateral parotitis beginning the day of the delivery. The child was subsequently severely ill and suffered from fever, splenomegaly and thrombocytopenia, however, without parotitis nor pancreatic involvement. Both mother and child recovered well with symptomatic treatment. A review of the literature shows that clinical mumps is rare and usually benign in neonates. However, severe respiratory distress may occur. The recent appearance of mumps outbreaks in adolescents and young adults calls for a reinforcement of mumps vaccination and should prompt an immunological assessment of pregnant women after exposure. PMID- 8223806 TI - The effect of variations in positive end expiratory pressure on gas exchange in ventilated children with liver disease. AB - The effect of varying the positive and expiratory pressure (PEEP) level during mechanical ventilation has been assessed in ten children with liver disease, mean age 3.8 years. PEEP was increased 3 cmH2O above the child's original (baseline) PEEP level and then decreased either by 3 cmH2O below the baseline or to 0 cmH2O. In all ten children increasing the PEEP above the baseline improved oxygenation; in the group overall the median PaO2 increased from 90 mmHg to 97 mmHg (P < 0.01). In eight of ten children decreasing the PEEP level below the baseline resulted in a deterioration in oxygenation; in the group overall the median PaO2 decreased from 91 mmHg to 82 mmHg (P < 0.05). Changes in PEEP levels, however, did not result in clinically significant alterations in PaCO2, heart rate or blood pressure. We conclude that modest increases in PEEP are well tolerated in children with liver disease and result in an improvement in oxygenation. PMID- 8223807 TI - Brain abscesses in neonates--report of three cases. AB - We report three newborns with brain abscesses. Two infants suffered from Serratia marcescens meningitis and one infant had enterococcal sepsis and meningitis. Brain abscesses were detected by cerebral sonography. Outcome in one infant with S. marcescens infection was poor. This patient developed multicystic encephalo malacia and severe developmental retardation. In the other patient with S. marcescens infection surgical drainage of the abscess was performed. The outcome was good both in this infant and in the patient with enterococcal brain abscess. PMID- 8223808 TI - Antipyretic effectiveness of acetaminophen in febrile seizures: ongoing prophylaxis versus sporadic usage. AB - A controlled clinical study compared the antipyretic effectiveness of acetaminophen administered at regular 4h intervals (group 1, n = 53) versus sporadic usage contingent upon a body temperature above 37.9 degrees C (group 2, n = 51) in 104 children presenting with simple febrile convulsions. The incidence of febrile episodes or temperature values were similar in spite of significantly larger amounts of acetaminophen administered to patients in group 1. Four and 4 children in groups 1 and 2, respectively, had a second episode of febrile seizures, in all of them within the first 24h of admission. We conclude that the prophylactic administration of acetaminophen in children with febrile seizures is not effective in the prevention of fever, the reduction of its degree, or in preventing the early recurrence of febrile seizures. PMID- 8223809 TI - Magnetic resonance imaging in juvenile Canavan disease. AB - We present a 2-year-old boy and a 6-year-old girl with mild Canavan disease (CD). Aspartoacylase activity in skin fibroblasts was deficient. Magnetic resonance imaging (MRI) of the brain did not show the prominent leucodystrophy previously reported in CD, but there was a hyperintense signal from the lentiform nuclei and the heads of the caudate nuclei on the T2-weighted MR images. This suggests a specific vulnerability of the corpus striatum in these patients. In the older patient, the white matter became affected at the age of 6 years. Proton magnetic resonance spectroscopy (1H-MRS) of white matter revealed a normal concentration of N-acetyl-L-aspartate (NAA) and a markedly decreased concentration of choline containing compounds (Cho) in the boy but a normal ratio of NAA to Cho in the girl. We conclude that deficient NAA catabolism affects myelin metabolism. This may present as changes in the striatum and/or as a low concentration of Cho before leucodystrophy appears on MRI. PMID- 8223810 TI - Light therapy as treatment of dyschronosis in brain impaired children. AB - Fourteen children aged 9 months-4 years with moderate to severe mental retardation and varying neurologic lesions were referred for severe and continuous nocturnal sleep disturbances and very abnormal day/night schedules. All children had previously been given hypnotic medications and behavioral treatments which had little or no effect on nocturnal sleep. The severity of the sleep disturbances significantly affected family life and was a major handicap to the children. All children were treated with light therapy (minimum 4000 lux). Five children responded to treatment and had normal sleep-wake cycles at the most recent post-treatment evaluation (2-5 years after the first treatment). Two of the patients' families were unable to follow the prescribed regimen. Treatment failed in 7 children. One of these seven children spontaneously improved 3 years later. In three of the failure children the neurologic problem progressively worsened, leading to death in one of them. Phototherapy is a treatment worth pursuing in children with very significant sleep/wake disruption which is unresponsive to behavioral or other treatments. It has few side-effects and may lead to normalization of the sleep-wake cycle. Recent improvement in the technology used to monitor the 24-h temperature rhythm over several days and the present commercial availability of "light boxes" should render these therapeutic trials easier than at the time of these initial investigations. PMID- 8223811 TI - Allergenicity of milk protein hydrolysate formulae in children with cow's milk allergy. AB - Cow's milk protein hydrolysate formulae have been developed to lower or eliminate the allergenicity of cow's milk proteins, and to reduce the antigenic load and the risk of sensitization. Cross-reactivity between different hydrolysate formulae and cow's milk proteins has been demonstrated. We have studied 20 children (median age 31 months, range 15-76 months) with a history of IgE mediated cow's milk allergy. All the children had immediate allergic respiratory and/or cutaneous and/or gastro-intestinal reactions to cow's milk ingestion. In addition, the children had positive prick skin tests and positive RAST to cow's milk. Prick skin test, RAST, and double-blind placebo controlled food challenges were performed with three different hydrolysate formulae: a casein hydrolysate formula and two whey formulae, one partially and one extensively hydrolyzed. All 20 children had immediate allergic reactions after the challenge test with cow's milk. Only 2/20 children had a positive challenge test with a casein hydrolysate formula (Alimentum): one developed asthma and one urticaria. Two of the 15 children challenged with an extensively hydrolysed whey formula (Profylac) developed perioral erythema. Nine out of 20 children had a positive challenge test with a partially hydrolysed whey formula (Nidina H.A.): four developed asthma, three urticaria and two lip oedema. All children had positive prick skin tests to cow's milk proteins (casein and/or lactalbumin); 9 to Nidina H.A.; 3 to Profylac, and 3 to Alimentum. Specific IgE antibodies to cow's milk were present in all children; in 13 to Nidina H.A., in 4 to Profylac, and in 3 to Alimentum. PMID- 8223812 TI - Congenital diaphragmatic hernia presenting after the newborn period. AB - Late-presenting congenital diaphragmatic hernia (CDH) is often difficult to diagnose and delay in treatment is common. Seven patients were operated beyond the newborn period for left-sided Bochdaleck hernia. Their age ranged from 1 month to 9 years. Six of them became symptomatic within the 1st year of life (1 week to 9 months of age). Either feeding difficulties or recurrent respiratory infections were initially present. In all of them chest X-rays were performed but delay in diagnosis ranged from 1 week to 5 years. All diaphragmatic defects could be closed by an abdominal approach without postoperative complications. Clinical symptoms disappeared postoperatively. In children with respiratory complaints or feeding difficulties one should be aware of late presenting CDH. A careful analysis of chest films and searching for "connecting" bowel segments passing through the diaphragmatic defect may help to avoid incorrect diagnosis and undesirable delay in treatment. Confusion with pneumonia or pneumothorax can be diminished by placing a feeding tube and instillation of contrast material. Ultrasound should be used supportively in all suspected diseases of the diaphragm. PMID- 8223813 TI - Carbamazepine-induced thrombocytopenia and leucopenia complicated by Henoch Schonlein purpura symptoms. AB - A rare case of carbamazepine-induced leucopenia and thrombocytopenia complicated by Henoch-Schonlein purpura (HSP) symptoms is presented. Laboratory findings suggested that leucopenia and thrombocytopenia could be due to bone marrow suppression and HSP symptoms to an allergic reaction to carbamazepine. To the best of our knowledge this is the first report that carbamazepine may cause haematological disorders associated with symptoms of HSP by different mechanisms at the same time in the same patient. PMID- 8223814 TI - Risk factors for childhood asthma and recurrent wheezy bronchitis. AB - Using cross-sectional data of an epidemiological study, risk factors for asthma and recurrent wheezy bronchitis were investigated in 1812 primary school children. Children with asthma (n = 63) had a similar pattern but a higher frequency of chronic respiratory symptoms than those with recurrent wheezy bronchitis (n = 136). Logistic regression analyses showed similar risk factors for both disorders, however, more pronounced for asthma. Prematurity was a significant risk factor for asthma and for recurrent wheezy bronchitis. Children with asthma more often had a family history of paternal or maternal asthma and their mothers tended to be younger. Effects of paternal asthma and prematurity were also found when the atopic status of the child (defined as skin test positivity to any of seven aero allergens) was taken into account. Next to genetic effects, adverse circumstances in early life seem to be important for the development of asthma. In school children recurrent wheezy bronchitis and asthma seem to be similar disorders which differ in quantitative but not qualitative aspects. PMID- 8223815 TI - Squamous-cell carcinoma arising in a non-irradiated child with recurrent respiratory papillomatosis. AB - We describe a patient with recurrent respiratory papillomatosis (RRP) associated with human papilloma virus (HPV), who developed a fatal squamous cell carcinoma of the lung. At the age of 1 year he presented with hoarseness, dyspnoea and inspiratory stridor but the diagnosis of RRP was made only 1 year later. At the age of 4 years he was tracheostomized because of upper airway obstruction. In spite of multiple surgical excisions and topic treatment with 5-fluorouracil the papillomata extended to the lung parenchyma. At the age of 16 years he developed a squamous-cell carcinoma of the lung and died 4 months later. Transformation to pulmonary carcinoma is a rare complication in non-irradiated patients with lung papillomatosis. We found only 11 similar cases in the literature. PMID- 8223816 TI - Medical errors during cardiopulmonary resuscitation of neonates. PMID- 8223817 TI - Early intussusception in premature infants. PMID- 8223818 TI - Anaphylactoid purpura and renal disease: the case against routine steroids. PMID- 8223819 TI - Early prednisone treatment and nephropathy in anaphylactoid purpura. PMID- 8223820 TI - Eosinophilic meningoencephalitis due to Toxocara canis. PMID- 8223821 TI - Elevation of transaminases as an early sign of late-onset glycogenosis type II. PMID- 8223822 TI - How physicians appraise everyday memory failures of patients across the adult life span. AB - A person-perception paradigm was used to investigate how primary care physicians appraise young, middle-aged, young-old, or old-old hypothetical patients (targets) who experience incidences of everyday memory failure. The physicians distinguished among targets who experienced short-, long-, and very long-term types of failure. However, their ratings of the point at which they would recommend intervention and of the referrals they would make in response to the three types of failure were not affected by the age of the target. The physicians' responses are compared with prior research results obtained for lay subjects' appraisals of young versus older targets' memory failures. The implications for primary care physicians' views of the significance of everyday memory failures in older adults are discussed. PMID- 8223823 TI - Stroop interference: aging effects assessed with the Stroop Color-Word Test. AB - A large, cross-sectional aging investigation of performance on the Stroop Color Word Test (SCWT) was carried out. Subjects were 247 volunteers, ages 20-80 in seven age levels. Although all subjects thought themselves to be normal and healthy, a post hoc division could be made on the basis of biological life events (BLE). BLE are mild biological or environmental factors, such as repeated experiences of general anesthesia, that can hamper optimal brain functioning. Apart from the anticipated age effects, performance was poorer in subjects who had experienced one or more BLE: The slowing due to BLE was comparable to the effect of age, especially on the task involving language interference in color naming. Education had a significant effect on performance: More highly educated subjects performed better than less educated subjects. No sex differences were observed. These findings replicate observations made with other tests in parallel studies. They are also in line with several other studies reporting interactions between the effects of aging and physical fitness. This study questions some of the validity of cognitive aging research, as our data suggest that screening for BLE as age-extrinsic factors in nondiseased subjects can reduce many of the performance deficits usually ascribed to aging per se. PMID- 8223824 TI - Leuko-araiosis and cerebral perfusion in normal aging. AB - To clarify the incidence, age relationships and pathogenesis of white matter lesions of unknown origin (leuko-araiosis) detected by neuroimaging among normal elderly volunteers, we measured the severity of leuko-araiosis using computerized tomographic (CT) densitometry among 42 healthy self-supporting men and women of different ages, all with normal neurological and cognitive test performance. Results were correlated with local cerebral perfusion using xenon-contrasted CT. The 42 volunteers, who are followed in this laboratory for studies of normal aging, were divided into two groups in order to determine aging effects by an extremes design. One group consisted of 19 adults below age 60 (M = 53.3, SD = 6.0). The index group comprised 23 individuals all over the age of 60 (M = 71.6, SD = 8.7). Leuko-araiosis around the anterior horns of the lateral ventricles (frontal leuko-araiosis) was more severe (p < .01) among the older group, however, occipital leuko-araiosis did not significantly differ between older and younger groups. Cerebral perfusion in frontal, temporal, and parietal cortex was decreased among older compared with younger volunteers (ps < .001, .01, and .05, respectively). Multiple regression analyses disclosed significant and direct relationships between severity of frontal leuko-araiosis and (a) frontal cortical atrophy and (b) reductions of cerebral perfusion within frontal white matter and caudate nucleus. We conclude that cortical atrophy with hypoperfusion and ischemia of frontal white matter play a part in the pathogenesis of frontal leuko araiosis associated with normal aging and this may be a predictor for later cognitive declines. PMID- 8223825 TI - Age and gender differences in the content scales of the Minnesota Multiphasic Personality Inventory. AB - We examined time of measurement, gender, and age differences on the nine content scales of the Minnesota Multiphasic Personality Inventory using data collected by three separate studies during the 1950s, 1960s and 1980s. No evidence was found for differences in the content scales due to time of measurement that also could not have been explained by demographic differences. Differences due to gender were found on only one of the nine scales, Masculinity-Femininity, and age differences were found on the Neuroticism, Extraversion, and Agreeableness scales. Younger men and women had significantly higher scores on the Neuroticism and Extraversion scales, and these differences were consistent in both magnitude and direction across sample and gender. Our results suggest that it is likely that openness reaches its lifetime stable level by the time typical adolescents enter college, because we found no significant age differences in intellectual interests. Neuroticism, extraversion, and agreeableness on the other hand, are likely to show instability throughout, and probably after, adolescence and early adulthood, because we found significant age differences in the content dimensions associated with these factors in separate analyses of three samples. PMID- 8223826 TI - Impact of age and situation-specific experience on maze learning and learning sensitive open-field parameters in mice. AB - The effects of age and situation-specific experience on maze learning and learning-sensitive open-field parameters (OFP) in Swiss albino male mice were studied. We tested the mice longitudinally by means of repeated measures and in cross-section by taking independent samples between the ages of 1-16 months. Mice with situation-specific experience improved their performance up to the age of 11 months. For mice lacking prior experience there was a trend toward impaired performance with regard to error scores and running times with increasing age. The ages of 1, 8, and 10 months were most favorable for the occurrence of quantitative maze learning-sensitive OFP changes, and the ages of 1, 3, 4, 8, 9, 10, 12, and 13 months were most favorable for the appearance of qualitative changes. The effect of situation-specific experience was interpreted as indicative of decreased emotionality but increased reactivity to novel stimuli or enhanced territorial behavior at the age of 16 months. PMID- 8223827 TI - Memory training in normal and demented elderly populations: the E-I-E-I-O model. AB - An heuristic classification scheme for memory interventions is described as a 2 x 2 matrix. One dimension of the matrix addresses the use of internal versus external mnemonics, and the other dimension addresses the use of explicit (effortful/conscious) versus implicit (automatic/unconscious) learning. Most attempts at memory interventions in normal older populations, such as the use of mental imagery, fall within the explicit-internal cell of the matrix and require expenditure of large amounts of cognitive effort. As a result, memory interventions in normal older populations, even when "successful," usually produce results that are limited in their generalization and are usually not maintained over time. Such interventions are rarely attempted in demented populations. Interventions are described that require less effort and are representative of other cells in the classification matrix. These may be more effective for demented and normal elders. Information acquired through these alternative means may eventually be stored internally and become consciously available, often producing an "O!" or "aha" experience when the individual realizes a shift to internal, consciously available storage has taken place. PMID- 8223828 TI - Beta-blockade and lipolysis during endurance exercise. AB - Inhibition of adipose tissue lipolysis may be involved in the impairment of endurance capacity after administration of a beta-adrenoceptor blocker. During endurance exercise, no significant decrease in plasma glycerol and free fatty acid (NEFA) concentrations after beta-adrenoceptor blockade is found. However, the levels during recovery from exhaustion are lower after beta-adrenoceptor blockade. This study was designed to investigate whether the lower levels after exercise are due to beta-adrenoceptor blockade or to the shorter time to exhaustion after administration of a beta-adrenoceptor blocker. In a single-blind study, 11 well-trained male subjects (age 23 (0.9) y) performed a cycle ergometer test at 70% Wmax until exhaustion 2 h after intake of 80 mg propranolol. One week later, the test was repeated after intake of placebo and was stopped at the time of exhaustion in the previous test. Average exercise time was 24 min. During exercise plasma glucose was lower, whereas plasma lactate and the respiratory exchange ratio were significantly higher when the subjects were on propranolol. Glycerol and NEFA concentrations during exercise were not significantly different between the two conditions. Despite an identical exercise time, glycerol and NEFA concentrations during recovery were significantly lower after propranolol treatment. In conclusion, lipolysis is inhibited during exercise after propranolol, probably causing a shift from fat to carbohydrate combustion. PMID- 8223829 TI - Amlodipine in patients with angina uncontrolled by atenolol. A double blind placebo controlled cross over trial. AB - Dihydropyridine calcium antagonists are established second line treatment for angina uncontrolled by beta-adrenergic blockers. Amlodipine is a recently introduced, dihydropyridine with a long half life. In a double blind, placebo controlled, cross over trial we assessed the efficacy and safety of amlodipine in 20 patients with persistent angina despite treatment with atenolol. 17 male patients (mean age 58 y) completed the study. Two patients were withdrawn during placebo because of worsening angina and one withdrew whilst on amlodipine because of palpitations. Compared with baseline, amlodipine prolonged exercise time to S T segment depression by a median of 12.5%; significantly more than was found with placebo (median 0%). The improvement in exercise time and time to angina also tended to be greater for amlodipine than placebo. GTN consumption, at a median of 1.3/week, was significantly less with amlodipine than placebo (2.8). Attacks of angina were also reduced. Standing systolic and diastolic blood pressures and sitting systolic blood pressure were lower with amlodipine than placebo. Heart rate did not change. There was no change in cardiac output (measured by doppler aortovelography) when amlodipine was added to atenolol. Holter monitor measurements of 24 h maximum and minimum heart rate, heart rate variation and extrasystole counts were the same for amlodipine and placebo. In conclusion, amlodipine is effective in patients with angina inadequately controlled by atenolol alone, and does not interfere with cardiac rhythm or function. PMID- 8223830 TI - Effects of a new oral hypoglycaemic agent, repaglinide, on metabolic control in sulphonylurea-treated patients with NIDDM. AB - We have evaluated the effects of repaglinide, a new non-sulphonylurea oral hypoglycaemic agent that has a stimulatory effect on insulin secretion. Forty four patients with NIDDM, already treated with a sulphonylurea, took part in an open, randomised, group comparison study of 12 weeks duration, during which they received either repaglinide or glibenclamide twice daily. While glibenclamide had a greater effect on fasting blood glucose (10.4 to 8.6 mmol.l-1), repaglinide significantly lowered postprandial blood glucose (13.8 to 12.2 mmol.l-1). Glycosylated haemoglobin remained unchanged in both groups, and serum fructosamine showed a tendency to fall. With both treatments total cholesterol was significantly decreased after 12 weeks, while HDL-cholesterol and triglycerides did not change. Fasting plasma insulin in the repaglinide group decreased from 80 (median value) to 67 pmol.l-1; it did not change in the glibenclamide group. Two patients in the repaglinide group did not complete the study, one for personal reasons, and one because of a rise in blood glucose. No abnormal findings attributable to repaglinide were observed in clinical and laboratory examinations, and no hypoglycaemic symptoms caused by it were observed. PMID- 8223831 TI - Intraduodenal infusion of a water-based levodopa dispersion for optimisation of the therapeutic effect in severe Parkinson's disease. AB - Motor performance of five patients with advanced Parkinson's disease was investigated during their optimum oral therapy (conventional tablets and/or depot capsules) and during a continuous duodenal infusion of levodopa. Due to the low water solubility of the drug, conventional tablets of levodopa+carbidopa (Sinemet) were milled and dispersed in a 1.8% aqueous methylcellulose solution. The dispersion was delivered nasoduodenally by a portable pump. The effect of levodopa in the two dosing regimens was estimated optico-electronically every 15 min and was also evaluated from videorecordings every 30 min and plasma levels of levodopa was regularly measured. Each dosage regimen the was studied twice, at a 2-4 day interval. Duodenal infusion improved motor function in all five patients and the fluctuations were reduced when compared to the oral therapy. Variation in plasma levodopa concentrations was 3-10 fold during oral therapy, while during the infusion a stable concentration was obtained. The therapeutic concentration varied from 0.3-3 ml-1 between patients. The relative bioavailability of levodopa in the solid preparation compared to the dispersion was in all patients 100%. Our results encourage further development of a duodenal infusion system with a levodopa dispersion for clinical use in parkinsonian patients who show severe fluctuation. PMID- 8223832 TI - Predictive performance of population pharmacokinetic parameters of tianeptine as applied to plasma concentrations from a post-marketing study. AB - The predictive ability of population pharmacokinetic parameters of tianeptine, obtained from a mixed effect analysis of pre-marketing pharmacokinetic studies, was evaluated using tianeptine plasma concentrations obtained during a large multi-center post-marketing surveillance study. The mean prediction error was 7.8 ng.ml-1 and the root mean square prediction error was 52.1 ng/ml when initial estimates of population pharmacokinetic parameters were used to predict drug concentrations in one half of the post-marketing data. When the population parameters were revised to reflect the data collected in the first half of the post-marketing study, the mean prediction error was reduced to -3.2 ng.ml-1 and the root mean square prediction error was reduced to 29.5 ng.ml-1. These results suggest that population pharmacokinetic parameters obtained from pre-marketing data may not accurately predict drug concentrations in patients receiving the drug in the post-marketing setting. Once the population parameters are updated to reflect data from the post-marketing period, the predictive ability of the data base increases, but substantial variability in the prediction error remains. PMID- 8223833 TI - Pharmacodynamics and pharmacokinetics of oral nitrendipine solution in hypertensive patients with advanced renal failure. AB - Nitrendipine solution 5 mg.ml-1 in the dose of 5 mg was given orally to 20 patients with chronic renal failure and elevated diastolic blood pressure (> or = 110 mmHg), of whom 10 were on maintenance haemodialysis (endogenous creatinine clearance < 5 ml.min-1) and 10 were at the predialysis stage (endogenous creatinine clearance 5-20 ml.min-1). The aim of the study was to investigate the influence of kidney function and/or dialysis treatment on the pharmacokinetic and pharmacodynamic profile of a solution of nitrendipine and to assess its antihypertensive efficacy. After 10 min there was a significant reduction in blood pressure from 188/113 to 173/100 (patients not dependent on dialysis) and from 197/112 to 161/94 mmHg (patients dependent on dialysis). The maximum fall in blood pressure (approximately 30%) was attained after 90 min in the dialysis patients and after 120 min in the non-dialysis group. Blood pressure increased again about 3 h after the administration of nitrendipine but it was still below baseline after 12 h. The terminal elimination half-life (4.1 h in the dialysis patients and 3.6 h in non-dialysis patients) was similar to that observed in patients with normal renal function. The pharmacokinetics of nitrendipine did not differ between the dialysis and non-dialysis groups. There was a correlation between plasma concentration and the blood pressure reduction. The maximum plasma concentration of nitrendipine was reached after 0.5 h (median) and did not differ between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223834 TI - Changes in renal clearance of furosemide due to changes in renal blood flow and plasma albumin concentration. AB - We have shown that, within therapeutic plasma concentrations, the unbound fraction of furosemide changes in direct proportion to the reciprocal of the plasma albumin concentration (correlation coefficient 0.99). Changes in the albumin concentration were produced by ultrafiltration of human plasma using a haemofiltration filter. Thus, we propose that, when studying changes in the pharmacokinetics of a highly protein bound drug, calculated changes in the unbound fraction offer an alternative to actual measurement of the unbound concentration, which is often difficult. Nine healthy volunteers receiving a continuous furosemide infusion were studied in normovolaemia and after dehydration (-1.4 kg), with and without pretreatment with an angiotensin converting enzyme inhibitor (captopril) or an a1-adrenoceptor blocking agent (prazosin). Significantly larger changes in the renal clearance of furosemide were found that could be explained by changes in the unbound fraction. Following dehydration, the unbound fraction of furosemide was decreased by about 5%, while its renal clearance fell by 27%, 33% and 13% after pretreatment with placebo, captopril and prazosin, respectively. The secretory clearance of the unbound furosemide changed substantially and in parallel with changes in the renal blood flow. It is suggested that changes in the renal clearance and excretion of furosemide and its t1/2 are much more dependent on changes in renal blood flow than on changes in its unbound fraction. PMID- 8223835 TI - The disposition of paracetamol and its conjugates during multiple dosing in patients with end-stage renal failure maintained on haemodialysis. AB - The disposition of oral paracetamol (1.0 g 3 times daily for 10 days) was studied in 6 patients with end-stage renal failure (creatinine clearance < 5 ml x min-1) maintained on haemodialysis 2 or 3 times per week. Blood was sampled daily for 10 days. The time of sampling depended on whether the patients were dialysed in the morning or afternoon but was always within 5 h of the last dose of paracetamol. On dialysis days samples were taken at the start of the session. The mean plasma concentration of paracetamol was 6.8 mg x l-1 after the first 24 h and subsequently varied little throughout the 10 days. Apparent steady-state plasma concentrations of 60.0 mg x l-1 and 54.5 mg x l-1 were reached for the glucuronide and sulphate conjugate of paracetamol respectively by the 2nd day of treatment with little variation throughout the remainder of the study. These steady-state concentrations of paracetamol glucuronide and sulphate were much lower than predicted. The steady-state plasma concentrations of the retained cysteine and mercapturate conjugates of paracetamol were low (5.7 and 3.7 mg x l 1, respectively) and there was no evidence of accumulation of these potentially toxic metabolites. It is not clear why regular dosing with paracetamol in haemodialysis patients did not cause the accumulation of paracetamol glucuronide or sulphate as predicted. There may be enterohepatic elimination of retained paracetamol conjugates or depletion of substrates such as inorganic sulphate during chronic dosing. PMID- 8223836 TI - Bambuterol and terbutaline in human cerebrospinal fluid and plasma. AB - Concentrations in cerebrospinal fluid (CSF) and plasma of bambuterol and its active metabolite, the beta 2-adrenoceptor agonist terbutaline, were measured in man after four once-daily doses of 30 mg bambuterol hydrochloride (Bambec). Nine patients scheduled for orthopaedic surgery under spinal anaesthesia completed the study. The concentrations of both substances were much lower in CSF than in plasma, the ratio CSF/plasma being 0.09 for bambuterol and 0.19 for terbutaline, at apparent steady state. While the rank order of the ratios was expected from the fractions of unbound bambuterol and terbutaline in plasma, their absolute values were only about 1/6 (bambuterol) and 1/4 (terbutaline) of those predicted from diffusion equilibria between plasma and CSF. Thus, the rates of transport of bambuterol and terbutaline from plasma into the central nervous system appear to be slow relative to transport out of the system, e.g. by outflow of CSF. The findings are in agreement with animal experiments and suggest that bambuterol and terbutaline are less likely than lipophilic beta 2-adrenoceptor agonists to interact with central receptors. PMID- 8223837 TI - Changes in the effects of nizatidine and famotidine on cardiac performance after pretreatment with ranitidine. AB - This was an open, randomized study of the cardiovascular effects of the histamine H2 receptor antagonists ranitidine, famotidine, and nizatidine after single oral doses alone or in combination in healthy volunteers. When compared with placebo ranitidine (450 mg) did not have any haemodynamic effects. Nizatidine (300 mg) caused significant falls in heart rate and cardiac output. Famotidine (40 mg) caused significant falls in stroke volume and cardiac output and an increase in pre-ejection period. Pretreatment with ranitidine abolished the haemodynamic effects of nizatidine and caused a time-shift of 2 h in the onset of the cardiovascular effects of famotidine. The difference in the results for nizatidine and famotidine can be explained by the longer half-life of famotidine. Vascular effects are assumed to be responsible for impairment of cardiac performance by famotidine. PMID- 8223838 TI - Dose-effect relationship of rilmenidine after chronic administration. AB - The antihypertensive efficacy and acceptability of 3 doses of rilmenidine (0.5, 1 and 2 mg, once daily) and a placebo over a 4 week period have been compared in a randomised, double-blind, parallel-group trial in 60 mild to moderate hypertensive patients. Six patients dropped out: 4 in the 2 mg-group and one in the 1 mg-group because of adverse events, and one in the placebo group for personal reason. The blood pressure was significantly decreased after the 1 and 2 mg doses with the maximum antihypertensive effect already being obtained after 1 mg. A significant dose-effect relationship was shown for supine systolic blood pressure (P = 0.05) but not for the supine diastolic blood pressure. The most beneficial efficacy/acceptability ratio was achieved at the dose of 1 mg once daily, which demonstrated the maximum antihypertensive effect associated with a low incidence of adverse events. PMID- 8223839 TI - The pharmacokinetics and pharmacodynamics of metoprolol after conventional and controlled-release administration in combination with hydrochlorothiazide in healthy volunteers. AB - We have studied a controlled-release formulation containing metoprolol 100 mg and hydrochlorothiazide 12.5 mg. We compared the pharmacokinetics of both substances and the pharmacodynamics of metoprolol with those of a conventional combination tablet. The controlled-release formulation gave less variable plasma metoprolol concentrations, Cmax 138 nmol.l-1 and Cmin 74 nmol.l-1, whereas for the conventional formulation the mean Cmax of metoprolol was 629 nmol.l-1 and the Cmin 20 nmol.l-1. Despite lower relative systemic availability (68%) for metoprolol from the controlled-release formulation and a smaller AUC, metoprolol from the controlled-release formulation produced a greater total effect, calculated as the area under the curve of the effect on exercise heart rate vs. time (303 vs. 259%.h; P < 0.05). Hydrochlorothiazide was rapidly absorbed from both formulations and the plasma concentration profiles were almost superimposable. Controlled-release metoprolol with hydrochlorothiazide combines effective beta 1-adrenoceptor blockade for 24 h without affecting the pharmacokinetics of hydrochlorothiazide. PMID- 8223840 TI - The pharmacokinetics of clonidine in high dosage. AB - We have evaluated the pharmacokinetics of high doses of clonidine, as used in the prophylactic treatment of alcohol withdrawal syndrome, in 11 alcohol-dependent patients undergoing surgery for oesophagogastrectomy. Clonidine was given in a bolus of 150 micrograms followed by a continuous infusion. After a mean period of treatment of 9.2 (range 3 to 26) days and a mean dose of 0.72 (range 0.29 to 2.37) mg per day of clonidine the mean terminal half-life was 15.8 (range 9.9 to 23) h (n = 7). In order to compare initial and terminal half-lives of clonidine intraindividually, four patients were given a bolus of 150 micrograms followed 24 h later by a continuous infusion. The pharmacokinetics of clonidine were described by two exponentials, with a distribution half-life of 1.2 h and a terminal half-life of 14.6 h. After a mean period of 8.3 (range 2 to 15) days and a mean dose of 0.62 (range 0.15 to 1.82) mg per day the terminal half-life in these four patients was 15.6 (range 14.0 to 17.9) h. The relation between dosage and plasma concentration was linear. PMID- 8223841 TI - Development of a population pharmacokinetic database for tianeptine. AB - Two thousand three hundred and thirty five plasma concentrations of tianeptine from 112 patients enrolled in nine studies of tianeptine pharmacokinetics performed prior to the marketing of the drug were pooled for analysis using mixed effect modeling. Studies represented a combination of single dose and multiple dosing at steady-state. Tianeptine plasma concentration time data were fit to a two compartment model with first order absorption using the NONMEM computer program. The results of this analysis suggested that alcoholism is associated with significant increase in clearance (124% increase) and volume of the central compartment (161% increase). The volume of the peripheral compartment is significantly lower in women (31% decrease) and in depressed patients (59% decrease). The population mean (interindividual variability) clearance was equal to 0.17 l.h-1 x kg-1 (28.6%), the volume of central compartment was 0.13 l.kg-1 (60.4%), intercompartmental clearance was 0.07 l.h-1 x kg-1 (30.1%), volume of the tissue compartment was 1.17 l.kg-1 (28.3%), and the absorption rate constant was 0.63 h-1 (21.8%). The residual variability was approximately 30% at concentrations expected during clinical use of the drug. Because of the increased clearance, alcoholic patients would be expected to have significantly reduced concentrations during steady-state dosing. These population parameters provide a basis for developing initial dosing recommendations and for performing bayesian evaluations of drug concentrations obtained in post-marketing studies. PMID- 8223842 TI - Decreased chloramphenicol clearance in malnourished Ethiopian children. AB - The disposition of chloramphenicol and chloramphenicol monosuccinate has been studied in thirty-four Ethiopian children of varying nutritional status. After a single intravenous dose corresponding to chloramphenicol 25 mg per kg bodyweight, the plasma clearance of chloramphenicol monosuccinate was decreased only in severely malnourished children with kwashiorkor. Seventeen % of the dose (range 0 51%) was recovered in urine as intact prodrug, indicating incomplete and variable bioavailability of chloramphenicol. Compared to underweight children, on average marasmic and kwashiorkor subjects exhibited a 2- and 3-fold increase, respectively, in the AUC of chloramphenicol. Elevated AUCs could be traced to reduced hepatic clearance of the drug. The unbound fraction both of chloramphenicol and its prodrug were slightly elevated in serum from kwashiorkor subjects. The possibility of using a single point measurement of plasma chloramphenicol as a guide to individualized dosage are discussed. PMID- 8223843 TI - Acute haemodynamic effects of isradipine in hypertensive heart transplant recipients. AB - The acute haemodynamic effects of the new calcium antagonist isradipine have been studied in ten hypertensive cardiac transplant recipients. Isradipine 0.01 mg.kg 1 was given IV during right heart catheterization and noninvasively automatic blood pressure measurements. From a baseline blood pressure of 164/114 mmHg the maximum blood pressure reduction was 35 mmHg systolic, 30 mmHg diastolic, and mean 34 mmHg, due to a reduction in systemic vascular resistance, maximum 43%. Cardiac index increased 32%, mainly due to an increase in the stroke volume, as the heart rate rose only by 10%. Left ventricular filling pressure was slightly reduced. We conclude that the main effect of isradipine was a major reduction in left ventricular afterload, comparable to that previously found in other patient groups. PMID- 8223844 TI - Itraconazole prevents terfenadine metabolism and increases risk of torsades de pointes ventricular tachycardia. AB - Terfenadine, a nonsedating H1-selective antihistamine, is widely used in many countries. We report pharmacokinetic results in a patient who developed a prolonged QT-interval in ECG and symptomatic torsades de pointes ventricular tachycardia as a consequence of the interaction of itraconazole and terfenadine. Both drugs were taken in the recommended doses: terfenadine 60 mg b.d. and itraconazole 100 mg b.d. Terfenadine metabolism was delayed by itraconazole, leading to an increased level of unmetabolised terfenadine. Seven weeks after the cessation of itraconazole treatment, terfenadine was rapidly metabolized to its active metabolite and did not prolong the QT-interval when given as a single provocation dose (120 mg). The findings suggest that intraconazole in therapeutic doses inhibits terfenadine metabolism. It is also possible that unmetabolised terfenadine alone, without an increased level of its active metabolite, may cause torsades de pointes. The concomitant use of terfenadine and itraconazole (and ketoconazole) should be avoided. PMID- 8223845 TI - The acute effects of amiloride and potassium canrenoate on digoxin-induced positive inotropism in healthy volunteers. PMID- 8223846 TI - New strategies in drug development and clinical evaluation: the population approach. Commentary on an action for co-operative research. PMID- 8223847 TI - A double-blind comparison of the effects of carvedilol and captopril on serum lipid concentrations in patients with mild to moderate essential hypertension and dyslipidaemia. AB - We have studied 250 patients with mild to moderate essential hypertension (diastolic blood pressure 95-114 mmHg) and dyslipidaemia (high-density lipoprotein cholesterol (HDL-C) below 1.03 mmol.l-1, total cholesterol 5.17-9.05 mmol.l-1, and triglycerides 2.26-5.64 mmol.l-1) in a controlled double-blind, multicentre, parallel group trial. The patients took a fat-modified diet. After a 4-week placebo period, patients who continued to fulfil the selection criteria were randomly allocated to treatment with either carvedilol (a vasodilating beta blocker) 25-50 mg o.d. (n = 116) or captopril (an ACE inhibitor) 25-50 mg o.d. (n = 117) for 6 months. In both groups there were favourable effects on the serum lipids. The relative changes (medians) in the carvedilol and captopril group were respectively: increase in HDL-C by 11% and 8%, decrease in total cholesterol by 11% and 10%, in low-density lipoprotein cholesterol by 16% and 12%, and in triglycerides by 13% and 14%. Equivalence of the two treatments was confirmed for the target variable change in HDL-C at a significance level of 5%. Reductions in supine systolic/diastolic blood pressures were comparable in the two groups (carvedilol: 23/19 mmHg, captopril: 20/18 mmHg). The improvement in lipid metabolism in patients treated with carvedilol is probably due to its alpha 1 blocking properties. PMID- 8223848 TI - Frequent occurrence of in vivo clonal expansion of CD4- CD8- T cells bearing T cell receptor alpha beta chains in adult humans. AB - We have previously reported 2 cases of healthy men showing in vivo monoclonal expansion of mature CD4- CD8- alpha beta T cells. In the present study, an additional 3 adults were found to exhibit such an expansion, among a total 464 adult donors studied. These 5 individuals were otherwise physiologically normal, with no history of severe illness and autoimmune disease at the time of examination. To investigate the mechanisms of the clonal expansion, further characterization of the clonal cells was attempted. No apparent preference for usage of the T cell receptor beta chain variable region was observed in the clonal T cells. These clonal T cells showed lectin-dependent or redirected antibody-dependent cell-mediated cytotoxicities, whereas they could not lyse autologous lymphoblastoid cell lines. Failure of Fas antigen expression was not observed for any of these clones. These results suggest that clonal expansion of CD4- CD8- alpha beta T cells frequently occurs in the periphery without any T cell abnormalities. PMID- 8223849 TI - Number of interleukin-4- and interferon-gamma-secreting human T cells reactive with tetanus toxoid and the mycobacterial antigen PPD or phytohemagglutinin: distinct response profiles depending on the type of antigen used for activation. AB - The enzyme-linked immunospot (ELISPOT) assay has been proven to be an efficient and sensitive method for the enumeration of single cells secreting antibodies or cytokines. Here we have used this method to determine the number of interleukin-4 (IL-4)- and interferon-gamma (IFN-gamma)-producing cells in vitro secondary responses to tetanus toxoid (TT) and the mycobacterial antigen (purified protein derivative; PPD) or the mitogen phytohemagglutinin (PHA). PHA-induced IL-4 and IFN-gamma secretion was well correlated suggesting polyclonal activation of cells. This was not the case with the specific antigens, where PPD preferentially induced IFN-gamma- and very few IL-4-producing cells, while TT-induced both IL-4 and IFN-gamma. These differences are probably a reflection of the types of immunity the two antigens induce, mycobacteria preferentially inducing a cell mediated T helper type 1 (Th 1) type of immunity, while immunity to tetanus is an antibody-dependent, Th 2 type of response. In individuals recently boosted with TT, a significant increase in both IL-4- and IFN-gamma-producing cells in response to TT was seen at day 7 after boost, followed by decline. This was in contrast to what was seen in response to PPD where an increase of IFN-gamma producing cells after the TT boost at day 7 persisted for at least 14 days. These results suggest that after an in vivo boost both antigen-specific and nonspecific T cells are activated and that antigen-specific cells home to other organs and therefore may be difficult to demonstrate in the circulation. Our data show that the ELISPOT assay is a powerful tool for determining the frequency of cells secreting cytokines. The assay has several advantages over other assays since it is sensitive, measures the number of actually secreting cells, and avoids the problems of binding of cytokines to their cell-bound or soluble receptors. PMID- 8223850 TI - Contrasting efficacy of presentation by major histocompatibility complex class I and class II products when peptides are administered within a common protein carrier, self immunoglobulin. AB - Major histocompatibility complex (MHC) class I and II products are specialized to present antigens via different intracellular processing routes. Peptides originating from proteins in the cytoplasm can gain access to class I peptide binding grooves, most likely in the rough endoplasmic reticulum. Peptides from proteins in acidic endocytic vacuoles gain access to class II. It has been proposed that MHC class I products also can capture peptides from "exogenous" or noninfectious sources, and this assumption underlies the use of intact proteins as vaccines for CD8+ cytotoxic T lymphocytes. Here we describe quantitative information comparing the efficacy of peptide presentation from exogenous proteins by administering a class I- and II-restricted peptide within the same context, the CDR3 loop of the VH domain of a self immunoglobulin. Antigen presenting cells (APC), including primary dendritic cells, efficiently present an influenza hemagglutinin peptide from the immunoglobulin (Ig) carrier (50% maximal response at 10 nM Ig-HA) to an MHC class II-restricted T cell. In contrast, these same APC are unable to present an influenza nucleoprotein (NP) peptide from the same context (1 microM Ig-NP) to an MHC class I-restricted T cell. Ig-NP DNA transfectants do present the nucleoprotein viral peptide on class I. Thus, peptides within the complementarity-determining region loops of Ig carriers can be presented on class I or II MHC products, but the endocytic compartment, when offered MHC class I- and II-restricted peptides within the same carrier protein context, favors presentation by class II by at least 1000-fold. PMID- 8223851 TI - Identification of an endogenous mammary tumor virus involved in the clonal deletion of V beta 2 T cells. AB - Expression of V beta (beta-chain variable region) gene segments was investigated in the Mus m. domesticus DDO strain, which possesses a large genomic deletion encompassing 20 of the 29 V beta gene segments known in BALB/c. Stainings using V beta-specific monoclonal antibodies revealed that up to 60% of the peripheral T cells use 3 V beta gene segments. Variable frequencies of V beta 2 T cells were observed among DDO individuals. Segregation analyses of F2 crosses between V beta 2-deletor mice and mammary tumor virus (Mtv)-free mice led to the identification of a new endogenous Mtv, named Mtv-DDO, mediating V beta 2 T cell clonal deletion. Mtv-DDO structure is conserved with the exception of the carboxy terminal region as compared to other Mtv. Comparison between Mtv sharing the same V beta specificity and isolated from laboratory or wild mice confirms that a stretch of 11 amino acids, defined as the V beta-specific region, is required for the V beta-specific interaction. Limited substitutions in this region account for the shift of the Mtv specificity towards different V beta. PMID- 8223852 TI - A new infectious mammary tumor virus in the milk of mice implanted with C4 hyperplastic alveolar nodules. AB - We have previously characterized an infectious mouse mammary tumor virus [(MMTV(SW)] which induces a strong superantigen response in vivo. Here we describe the isolation and characterization of MMTV(C4) which was derived from milk of mice implanted with hyperplastic alveolar nodules. MMTV(C4) stimulates V beta 2 expressing T cells after local injection in vivo. Comparison with known open reading frame (orf) sequences revealed high homology to Mtv-6, an endogenous virus interacting with V beta 3-expressing T cells. The carboxyl-terminal amino acids were, however, altered. High homology including the carboxyl-terminal orf amino acids were found with MMTV(C3H-K). We show here that MMTV(C3H-K) has lost its superantigen function. Sequence comparisons permitted the characterization of few key amino acids which could be important for T cell receptor interaction and superantigen processing. PMID- 8223853 TI - Tumors with reduced expression of a cytotoxic T lymphocyte recognized antigen lack immunogenicity but retain sensitivity to lysis by cytotoxic T lymphocytes. AB - A murine solid tumor was transfected to express various levels of an allogeneic major histocompatibility complex class I gene (K216), in order to test the effect of the level of antigen expression on immunogenicity and sensitivity to lysis by cytotoxic T lymphocytes (CTL). The growth rates of clones of tumor cells expressing different levels of the transfected gene were similar in vitro and in nude mice. Although all tumor cells, including cells freshly isolated from growing tumors, were equally sensitive to lysis by specific CTL, only tumor cells expressing the highest level of the K216 antigen stimulated CTL and were rejected by normal mice. In contrast, tumor cells expressing lower levels of antigen failed to immunize for CTL and grew progressively in normal mice, despite retaining expression of the transfected gene and remaining fully sensitive to CTL mediated lysis; thus, the threshold of antigen needed to stimulate CTL responses was considerably higher than that needed to lyse tumor cells. Reduction of K216 antigen expression from 100-fold to 40-fold above background, impaired significantly the ability of the tumor cells to induce a K216-specific immune response, while tumor cells expressing K216 at levels 2-fold above background were as susceptible to CTL-mediated lysis as tumor cells expressing 50-fold more antigen. The important implication of these findings is that some tumors occurring in nature may not be immunogenic but nevertheless express antigens which are potential targets for immune therapy. PMID- 8223854 TI - Activation of human monocytes and granulocytes by monoclonal antibodies to glycosylphosphatidylinositol-anchored antigens. AB - The present study investigated possible receptor-like characteristics of glycosylphosphatidylinositol (GPI)-linked antigens on human monocytes and granulocytes by measuring cytoplasmic calcium fluxes and the oxidative burst in cells following cross-linking of GPI-linked antigens. Cross-linking of cell-bound anti-CD14, -CDw52 and -CD55 induced cytoplasmic calcium fluxes and oxidative bursts in unprimed human monocytes similar to those observed following Fc gamma R cross-linking. In granulocytes primed with 200 mM N-formyl-Met-Leu-Phe (FMLP), cross-linking of cell-bound anti-CD16, -CD24, -CD59 and -CD67 led to calcium fluxes and activation of the oxidative burst. The oxidative bursts mediated by GPI-linked antigens were stronger than those induced by 200 nM FMLP, even though FMLP induced a larger increase in cytoplasmic calcium concentration. The responses were likely to be independent of Fc gamma R interactions as F(ab')2 fragments of IgG or IgM antibodies were used in the experiments. Activating effects of monoclonal antibody to GPI-linked antigens were not observed in cells from patients with paroxysmal nocturnal hemoglobinuria, which are deficient in GPI-linked antigens. In addition, treatment with GPI-specific phospholipase C led to inhibition of cell activation through GPI-linked antigens but not through transmembrane receptors. Cross-linking of a number of non-GPI-linked antigens (CD11a, CD18, CD31, CD35, CD43, and CD45) neither induced calcium fluxes, nor activated the oxidative burst. The results indicate that most, if not all, GPI linked surface glycoproteins on myeloid cells are capable of mediating cell activation and suggest that the GPI anchor is a structure facilitating signal transduction. PMID- 8223855 TI - ICAM-3, the third LFA-1 counterreceptor, is a co-stimulatory molecule for both resting and activated T lymphocytes. AB - Optimal activation of human T cells mediated by ligation of CD3/T cell receptor (TcR) complex requires co-stimulatory signals. These can be provided by the adhesive interaction between receptor molecules on T cells and their counter receptors on antigen-presenting cells. Soluble ICAM-3, anti-ICAM-3 and anti-CD3 mAb were utilized to address the role of the ICAM-3/LFA-1 pathway in TcR/CD3 dependent or -independent T cell activation. Immunoaffinity-purified ICAM-3 co immobilized with suboptimal concentrations of anti-CD3 monoclonal antibody (mAb) stimulated T lymphocytes as monitored by the expression of the lymphocyte activation antigens CD25 and CD69. The mechanism underlaying this activation appear to involve the interaction of ICAM-3 with a beta 2 integrin, likely to be LFA-1, since mAb to the CD18 chain completely inhibited T cell activation. Similar experiments demonstrated that anti-ICAM-3 mAb were able to co-stimulate both resting (cord blood) and activated (T cell clones) T lymphocytes. On the contrary, anti-ICAM-1 mAb were only co-stimulatory for CD25 expression on activated but not on resting T cells. In addition, we have found that some gamma delta T cell clones bearing the V delta 1 segment were activated by direct mAb engagement of ICAM-3 in the absence of TcR/CD3 occupancy. Furthermore, immobilized anti-ICAM-3 mAb also induced development of dendritic processes. In conclusion, our data suggest that ICAM-3 on the surface of both T cells and antigen-presenting cells plays an essential role in the initiation of the immune response. PMID- 8223856 TI - Schistosoma mansoni circulating anodic antigen but not circulating cathodic antigen interacts with complement component C1q. AB - Adult schistosome parasites, living in the blood vessels of their mammalian hosts, protect themselves against immune damage in a variety of ways. In addition to the tegument, the intestinal epithelium of the blood-feeding worms is permanently exposed to both the innate and the acquired immune system. In this study, we investigated whether the Schistosoma gut-associated antigens CAA and CCA (circulating anodic antigen and circulating cathodic antigen, respectively), which are excreted in relatively large quantities into the host's circulation, might play a role in evading complement attack. Of several complement components tested, only purified C1q showed significant binding to CAA, a negatively charged highly glycosylated glycoprotein. CCA, also highly glycosylated, but neutral or slightly positively charged, did not bind to C1q. CAA bound only to the collagen like stalks of C1q and not to the globular heads. No detectable interaction of CAA with precursor human C1 was found and CAA did not induce activation of C1 in whole human serum as assessed by consumption of hemolytic C4 activity. Also CAA could not induce activation of precursor C1 in vitro. These results suggest that CAA behaves like a receptor for C1q, and might be involved in protecting the vulnerable schistosome gut against complement-mediated attack. PMID- 8223857 TI - Somatic diversification and affinity maturation of IgM and IgG anti-DNA antibodies in murine lupus. AB - Molecular events occurring during the process of generation of pathogenic immunoglobulin (Ig)G anti-DNA antibodies in systemic lupus erythematosus (SLE) were studied using a newly established method. We analyzed the Ig variable (V) region gene sequence and DNA-binding activity of IgM and IgG anti-DNA monoclonal antibodies (mAb) from individual SLE-prone (NZB x NZW) F1 mice. The first event appeared to be clonal selection and expansion of IgM anti-DNA clones, in which several clones had intraclonal V gene mutations. Although the number of mutations was small, the mutated IgM clones were associated with an increase in DNA-binding activity. The somatic mutations located in complementarity-determining regions (CDR) and in framework regions (FR) of V genes were apparently related to changes in DNA-binding activity. IgG anti-DNA clones that progressively increased in number with aging had numerous somatic mutations in the V region genes and there was a pair of clones which showed an intraclonal accumulation of mutations, in association with increase in the DNA-binding activity. All these findings show that somatic mutations associated with affinity maturation of the V region begin immediately before isotype-switching from IgM to IgG of the clones that have been selected and expanded, in an antigen-driven manner and/or by other forces. We propose that further accumulations of intraclonal somatic hypermutation, in association with selection and expansion of high affinity IgG clones, may lead to formation of highly pathogenic anti-DNA antibodies. PMID- 8223858 TI - Signaling of programmed cell death induction in WEHI-231 B lymphoma cells. AB - The murine WEHI-231 B lymphoma is highly sensitive to membrane immunoglobulin ligation which leads to programmed cell death (PCD) in this cell line. To study the molecular pathways involved in PCD induction in these cells, we derived two variants of WEHI-231 resistant to anti-Ig treatment. The level of bcl-2 mRNA was identical in the wild type and the variants, either untreated or anti-Ig treated, suggesting that PCD is not under the control of bcl-2 in WEHI-231 cells. In contrast, c-myc gene expression was markedly different in the wild type and the variants, both in the unstimulated and anti-Ig-stimulated state. Our findings are interpreted in the context of the dual capacity of c-myc to promote cell growth or cell death, in conjunction with other growth regulatory signals. PMID- 8223859 TI - Stimulation of the intracellular killing of Staphylococcus aureus by human monocytes mediated by Fc gamma receptors I and II. AB - Previous studies have shown that intracellular killing of bacteria by monocytes is stimulated by interaction between IgG and Fc gamma receptors (Fc gamma R) in the membrane of these cells. In the present study anti-Fc gamma R monoclonal antibodies (mAb) were used to investigate the relative contributions of the various classes of Fc gamma R to the intracellular killing of Staphylococcus aureus by human monocytes and the biochemical pathways involved. Anti-Fc gamma RI or anti-Fc gamma RII mAb, but not anti-Fc gamma RIII mAb, efficiently stimulated the intracellular killing of bacteria by monocytes. Cross-linking Fc gamma RI or Fc gamma RII, but not Fc gamma RIII, on monocytes with mouse anti-Fc gamma R mAb followed by bridging with F(ab')2 fragments of goat anti-mouse IgG enhanced this process. Since the NADPH oxidase inhibitor diphenyleneiodonium blocked the Fc gamma R-mediated intracellular killing of S. aureus, oxygen-dependent bactericidal mechanisms are most probably involved. Cross-linking Fc gamma RI or Fc gamma RII but not binding of the mAb to the Fc gamma R on monocytes activated phospholipase C, as demonstrated by the increase in the intracellular concentration of inositol-(1,4,5)-triphosphate. The enhanced intracellular killing stimulated by cross-linking Fc gamma R on monocytes was completely blocked by U-73122, an inhibitor of phospholipase C-dependent processes. Protein kinase C activity, but not the rise in the cytosolic free Ca++ concentration or pertussis toxin-sensitive G proteins, is essential for the Fc gamma R-mediated intracellular killing of bacteria by monocytes. Together, these results demonstrate that cross-linking Fc gamma RI or Fc gamma RII is equally effective in stimulating the intracellular killing of bacteria by monocytes and that this stimulation is a phospholipase C-dependent process. PMID- 8223860 TI - The lack of CD8 alpha cytoplasmic domain resulted in a dramatic decrease in efficiency in thymic maturation but only a moderate reduction in cytotoxic function of CD8+ T lymphocytes. AB - The glycoprotein CD8 is believed to play an important role in the maturation and function of MHC class I-restricted T lymphocytes. CD8 has been proposed to function as a co-receptor of the TcR to participate in signal transduction, possibly through its cytoplasmic domain that binds to protein tyrosine kinase p56lck. A T cell-specific transgene encoding CD8 alpha truncated at the cytoplasmic domain ("tailless CD8 alpha"), was introduced into CD8 alpha deficient mice. This animal model was used to study the role of the CD8 cytoplasmic domain in T cell ontogeny and function. "Tailless CD8 alpha" was expressed on the cell surface of thymocytes and peripheral T cells. A small population of peripheral CD4- T cells (6% of T lymphocytes) was found to have cell surface expression of "tailless CD8 alpha" and endogenous CD8 beta, indicating that these cells may belong to the CD8+ T cell lineage. A consistent result was obtained from CD8 alpha-deficient mice bearing the "tailless CD8 alpha" and the MHC class I-restricted 2C TcR transgenes. A small population of CD4- T cells expressing CD8 beta, the "tailless CD8 alpha" and the 2C TcR transgenes was present in the periphery of these mice in a selecting background, but was absent in a deleting background. When "tailless CD8 alpha" mice were infected with lymphocytic choriomeningitis virus (LCMV), the peripheral CD8+ CD4- T cell subset expanded dramatically and a significant LCMV-specific cytolytic activity was detected. The results suggest that the cytoplasmic portion of CD8 alpha is not absolutely required but dramatically enhances the efficiency of thymic maturation of CD8+ T cells. The lack of CD8 alpha cytoplasmic domain in peripheral CD8+ T cells does not abolish the generation of cytotoxicity in response to an in vivo LCMV infection, although the cytolytic activity is slightly reduced compared to that in control mice. PMID- 8223861 TI - Global analysis of antibody repertoires. II. Evidence for specificity, self selection and the immunological "homunculus" of antibodies in normal serum. AB - The serum IgM repertoires of C57BL/6, DBA/2 and BALB/c mouse strains were analyzed using a recently developed global and quantitative assay that measures antibody reactivities to a very large number of antigens. A characteristic repertoire could be assigned to each strain. The different repertoires could be successfully classified with multivariate statistics. Many common reactivities were also observed among the different strains, which allows the definition of a mouse-specific repertoire. Analysis of human sera support this notion. To investigate the impact of minor genetic differences on the serum IgM repertoire, the congenic strains B10.D2/oSn and B10.D2/nSn, which differ in the expression of the C5 component of complement, were analyzed. The two strains could be separated based on the reactivity profiles obtained. The analysis of the results reveals that many antigenic proteins are not recognized at all by natural antibodies, while others are disproportionately reactive, the resulting patterns giving rise to what could be the definition of an "immunological homunculus". The relevance of this type of analysis for clinical applications is discussed. PMID- 8223862 TI - The human immunoglobulin kappa locus. Characterization of the partially duplicated L regions. AB - The L regions are parts of the C kappa proximal (p) and distal (d) copies of the human immunoglobulin kappa locus and are therefore called the Lp and Ld regions. The two regions with their 25 V kappa genes and pseudogenes have now been cloned, thus completing the cloning of the kappa locus. Lp has been linked to the neighboring Ap and B regions, while Ld was linked to Ad. There is good evidence that at the other side of Ld, i.e. towards the centromere, the end of the locus has been reached. Most of the cloning and linking was achieved by chromosomal walking, employing cosmid and phage lambda clones. No such clones could be found for three small gaps. Two of them were closed by a polymerase chain reaction strategy; the third one was characterized by genomic blot hybridization experiments and eventually bridged by a yeast artificial chromosome clone. Early in evolution, a stretch of about 25 kb which comprised three V kappa genes near the 5' end of the L region precursor must have been duplicated, such that the later duplication of large parts of the kappa locus resulted in the appearance of two very similar three-gene regions in each, Lp and Ld. Two deletions in the central parts of the L regions, on the other hand, must have occurred after the duplication of the locus, since they are found in Lp and Ld in different positions. PMID- 8223863 TI - The V kappa genes of the L regions and the repertoire of V kappa gene sequences in the human germ line. AB - Only 14 of the 25 V kappa genes and pseudogenes had been found before as parts of the L regions. The cloning and linking described in the accompanying report allowed us now to assign to Lp or Ld some V kappa genes which had been found before on scattered clones. In addition the sequences of several still unknown genes are reported here, thus completing the publication of the V kappa genes of the kappa locus as far as they are potentially functional or have only one or two 1-bp defects. Of the V kappa genes of the kappa locus, 32 are potentially functional, 16 have minor defects, 3 have both potentially functional and slightly defective alleles and 25 are pseudogenes which amounts to a repertoire of 76 V kappa-related gene sequences. The V kappa genes of the L regions are, within the subgroups, particularly similar to each other, which is in part due to common evolutionary origins and in part caused by gene conversion-like events. One donor-acceptor pair could be clearly identified, since converted and not converted alleles of the acceptor gene were found. In other cases the duplicates of the converted genes served as non-converted controls. PMID- 8223864 TI - Patterns of major histocompatibility complex class II expression on T cell subsets in different immunological compartments. 1. Expression on resting T cells. AB - In this study we have investigated the expression of major histocompatibility complex (MHC) class II molecules on T cells from various lymphoid compartments in the sheep. Monoclonal antibodies which react specifically with sheep MHC class II molecules homologous to the human DQ and DR molecules have been characterized. These antibodies have been used, together with the monoclonal antibodies specific for sheep CD4-, CD8- and T19-positive T cells, to quantitate DQ and DR expression on T cell subsets in adult and fetal peripheral blood, afferent lymph, lymph node and efferent lymph. The results show that expression of class II by T cells depends on the age of the animal and the physiological location of the T cell. In fetal blood there is no expression of class II on CD8+ or T19+ cells and very low expression on CD4+ T cells. In adult peripheral blood and efferent lymph a significant proportion of cells express DR but not DQ. A very different situation is found in afferent lymph and the peripheral lymph node: in afferent lymph the majority of T cells in all three subsets express both DQ and DR molecules; in the lymph node over 50% of T cells express DR and 30% are DQ+. These results suggest that within all T cell subsets there is a progression from DQ-DR- to DQ-DR+ and DQ+DR+ which correlates with physiological stages of T cell differentiation in vivo. PMID- 8223865 TI - Patterns of major histocompatibility complex class II expression by T cell subsets in different immunological compartments. 2. Altered expression and cell function following activation in vivo. AB - This study characterizes antigen-induced phenotypic and functional aspects of major histocompatibility complex (MHC) class II expression on recirculating T cells in efferent lymph. In vivo secondary, but not primary challenge is associated with both kinetic and phenotypic alterations in class II expression by T cells. All three major T cell subsets, CD4+, CD8+ and T19+ (gamma delta T cell receptor), show an approximate four fold increase in the level of MHC class II expression during secondary responses. No changes in B cell expression of class II were seen. Resting efferent lymph T cells are predominantly either class II- or DR+DQ- but this changes to DR+DQ+ after antigenic challenge. The antigen presenting function of these class II+ T cells was investigated at daily intervals after in vivo antigenic challenge. T cells from non-activated lymph nodes could not induce proliferation of antigen-specific T cells with soluble antigen but were weakly stimulatory in allo-mixed lymphocyte reaction (MLR) at high (> 2:1) stimulator cell ratios. Activated T cells isolated during secondary in vivo responses, and expressing increased quantities of MHC class II, were positive stimulator cells in the MLR. In contrast these cells could not present soluble antigen or trypsin-digested antigen to the T cell lines. In the MLR assays, the relative stimulation by class II+ T cells correlates with the levels of class II expression. We conclude from these experiments that both quantitative and qualitative changes in MHC class II, induced on T cells under physiological conditions, play a role in the regulation of the immune response in vivo but that that role is not simply one of presentation of soluble antigen. PMID- 8223866 TI - Major histocompatibility complex regulation of interleukin-5 production in the mouse. AB - Lymph node cells of CBA (H-2k), but not of BALB/c (H-2d) mice immunized epicutaneously with picryl chloride secrete interleukin (IL)-5 when stimulated with the specific antigen in vitro. The low IL-5 production in BALB/c mice persists when either picryl chloride or the unrelated antigen oxazolone are used, when the amount of antigen in vitro is varied and when a secondary response is studied. The difference in IL-5 production maps to the major histocompatibility complex (MHC) in the congenic BALB/b, BALB/c and BALB/k mice. Furthermore, lymph node cells from (k x d) F1 mice produce IL-5 when stimulated by antigen presented on H-2k but not on H-2d antigen-presenting cells. Finally, the low IL-5 production in vitro in BALB/c mice is correlated with low picryl-specific IgA levels in vivo, which otherwise are ten times greater in CBA and BALB/k mice. The influence of MHC on IL-5 production and IgA secretion in the mouse might be a possible basis for the association of MHC with IgA deficiency in humans. PMID- 8223867 TI - Antisense oligonucleotides specific for regulatory factor RFX-1 inhibit inducible but not constitutive expression of all major histocompatibility complex class II genes. AB - The regulation of major histocompatibility complex (MHC) class II genes expression, which can be constitutive, inducible or both, is a crucial aspect of the control of an immune response. It involves binding of various regulatory factors to cis-acting sequences of MHC class II promoters. Antisense oligonucleotides specific for RFX-1, a regulatory factor binding to the functionally essential X box motive of MHC class II promoters, were designed to study the role of RFX-1 in the various modes of MHC class II regulation and explore the possibility of experimentally modulating the level of expression of MHC class II genes by transcriptional intervention. RFX-1 antisense oligonucleotides were first tested in cell-free translation, selected for an inhibitory effect on RFX-1 in vitro translation and then assayed in cell cultures for an effect on human histocompatibility leukocyte antigen (HLA) class II expression. We show that an RFX-1 specific antisense oligonucleotide drastically inhibits induction of HLA-DR,-DQ, and -DP molecules by interferon gamma in monocytic cells. Unexpectedly, the same agent has no effect on the constitutive expression of the same genes either in these cells or in B lymphocytes, indicating an uncoupling of the constitutive and inducible modes of class II regulation. This transient and reversible experimental modulation of MHC class II expression in live cells by transcriptional intervention provides a new tool to study the function of class II molecules in various biological models. PMID- 8223868 TI - T cell-mediated cognate signaling of nitric oxide production by macrophages. Requirements for macrophage activation by plasma membranes isolated from T cells. AB - Macrophage generation of reactive nitrogen intermediates (RNI) represents a major effector mechanism in anti-microbial immunity and non-septic inflammatory reactions. The induction of macrophage RNI production has been demonstrated to require at least two signals which in microbial infections can be provided by interferon (IFN)-gamma and lipopolysaccharide (LPS). The current study demonstrates that, in the absence of LPS, T lymphocytes can provide cognate signal(s) which synergize with IFN-gamma in stimulating macrophage RNI production, as evidenced by the ability of plasma membranes from T cell clones to activate IFN-gamma-primed macrophages. Although viable resting T cells can activate IFN-gamma-primed macrophages by an interaction that is antigen specific, plasma membranes from resting T cells do not active macrophages. Plasma membranes from T cells activated by immobilized anti-CD3 were able to effectively induce RNI production in IFN-gamma-primed macrophages. However, in contrast to the antigen-specific interaction of macrophages with viable resting T cells, the activation of IFN-gamma-primed macrophages by membranes from activated T cells does not display antigen specificity. Plasma membranes from activated T helper TH2 and from activated TH1 cells were equally effective in activating IFN-gamma primed macrophages, suggesting that the dominance of TH1 over TH2 cells in cell mediated responses involving macrophage effectors is not a reflection of differences in their ability to interact with macrophages but rather is a reflection of their different pattern of cytokine production. These results suggest that the T cell-macrophage interaction involves reciprocal activation of both cells--an antigen-specific activation of the T cells which results in the acquisition of T cell membrane components involved in antigen-nonspecific stimulation of the macrophages. PMID- 8223869 TI - L1 adhesion molecule on mouse leukocytes: regulation and involvement in endothelial cell binding. AB - L1 is a cell surface glycoprotein of the immunoglobulin superfamily which was initially shown to mediate adhesion between neural cells. Recently we have reported that L1 is expressed by bone marrow cells and the majority of mature lymphocytes (Kowitz et al., Eur. J. Immunol. 1992. 22: 1199-1205). To analyze the function of L1 on leukocytes we studied its regulation following cell activation. In vitro activation of B lymphocytes with lipopolysaccharide or T lymphocytes with phorbol 12-myristate 13-acetate/Ca2+ ionophore, concanavalin A or anti-CD3 monoclonal antibody as well as in vivo activation of V beta 8+ T cells with staphylococcal enterotoxin B (SEB) revealed a down-regulation of L1 within 48 h. A rapid loss of L1 expression was seen when mouse neutrophils were activated with PMA alone. This rapid loss paralleled the shedding of L-selectin. We also studied a possible role of L1 in the binding of leukocytes to endothelial cells. ESb-MP lymphoma cells with a high expression of L1 (L1hi) could bind to bend3 endothelioma cells without prior activation with inflammatory cytokines. The interaction was inhibited by anti-L1 antibodies. In contrast, ESb-MP cells with low L1 expression (L1lo) were only marginally bound. Latex beads coated with affinity-isolated L1 antigen were also able to bind to the endothelioma cells in a specific fashion. The binding of ESb-MP lymphoma cells required Ca2+ and Mg2+ ions and was sensitive to cold temperature. Since the endothelioma cells did not express L1 the binding mechanism studied here is distinct from the established L1 L1 homotypic interaction. It is possible that the novel L1-mediated adhesion pathway involves an unidentified ligand and could play a role in leukocyte migration. PMID- 8223870 TI - Effects of murine leukemia viruses on the function of dendritic cells. AB - In asymptomatic human immunodeficiency virus-1 infection T cells respond normally to allogeneic dendritic cells (DC), but DC show reduced stimulatory capacity. By contrast in HTLV-1 infection no significant changes in allogeneic stimulation were seen but DC-stimulated activity of autologous T cells. In seeking animal models relevant to these diseases the effects of two murine leukemia retroviruses, Rauscher leukemia virus (RLV) and Moloney leukemia virus (MLV) on the function of dendritic cells and T cells in a primary mixed leucocyte reaction have been tested. Treatment by RLV in vitro suppressed the ability of DC to stimulate allogeneic T cells from healthy animals. MLV at the same concentration did not significantly affect the ability of DC to stimulate allogeneic T cells, but provoked considerable enhancement of the low level stimulation by DC in the syngeneic system. Similar results were obtained following in vivo exposure to viruses. Two pieces of evidence suggested that these effects were due to impairment of DC function and were not operating through infection of T cells. Firstly, exposure of T cells directly to virus in vitro and in vivo before stimulation with untreated allogeneic DC caused no significant alteration in T cell activity. Secondly, the impact of murine leukemia virus on DC function was not abrogated when infected DC were added to normal T cells and cultured in the presence of zidovudine. Treatment of DC by RLV caused a decrease of cluster formation with allogeneic T cells. No statistically significant influence of MLV was observed on cluster formation after 3-h of incubation in the allogeneic system. However, after 18-h incubation MLV-treated DC formed fewer clusters with T cells than untreated DC. At the same time a stimulatory effect of MLV on DC cluster formation with syngeneic T cells was found. Considerable decrease was found in major histocompatibility complex class II antigen and LFA-1 receptor expression on the DC surface in mice infected by RLV. MLV induced no significant changes. These mouse retroviruses can therefore cause changes in DC function similar to those already reported using human retroviruses and may provide models for studying their effects. PMID- 8223871 TI - Cleavage and inactivation of human C1 inhibitor by the human leukocyte proteinase, proteinase 3. AB - Incubation of highly purified human C1 inhibitor with equally pure human leukocyte proteinase 3, resulted in a dose- and time-dependent inactivation of C1 inhibitor hemolytic activity. Furthermore, this inactivation was accompanied by proteinase 3-dependent cleavage of the C1 inhibitor into an 83,000 molecular weight fragment. The formation of the 83,000 molecular weight fragment followed a time course which was similar to that observed for the inactivation of hemolytic activity. Within 120 minutes more than 90% of the hemolytic activity was lost. This inactivation of C1 inhibitor appeared to be selective as purified human C1q was not degraded in a similar time period. Moreover, when 100 micrograms IgG, isolated from each of 21 Wegener's granulomatosis patients with cytoplasmic anti nuclear antibodies immunofluorescent titers to proteinase 3 greater then 1:64, was incubated with 3 milliunits of proteinase 3, little to no cleavage of C1 inhibitor was observed. In contrast, 100 micrograms of IgG isolated from 14 normal donors was ineffective in affording protection to C1 inhibitor upon incubation with proteinase 3. Our results suggest that neutrophil infiltration and activation could lead to local complement consumption at the tissue sites. PMID- 8223872 TI - Selection of the expressed B cell repertoire by infusion of normal immunoglobulin G in a patient with autoimmune thyroiditis. AB - In the present study we have analyzed the changes in the expressed antibody repertoire and in temporal fluctuations of antibody levels in serum that followed infusion of normal IgG (IVIg) in a patient with autoimmune thyroiditis. Administration of IVIg resulted in the stimulation of IgM production, in alterations of expressed antibody activity in serum that could not merely be accounted for by the passive transfer of antibody specificities contained in IVIg, in transient down-regulation of B cells clones expressing a specific disease-related idiotype and in the increase in serum in recipient's autoantibodies specifically reactive with F(ab')2 fragments of IVIg. In addition, infusion of IVIg shifted the pattern of spontaneous fluctuations of autoantibody activities in the patient's serum from a pattern indicative of disconnected events in the immune network to a pattern similar to that which is consistently observed in healthy controls. These results suggest that normal IgG may modulate autoreactivity by selecting expressed antibody repertoire through V region dependent interactions with antibodies. PMID- 8223873 TI - Interleukin-1 dysregulation is an intrinsic defect in macrophages from MRL autoimmune-prone mice. AB - Macrophages (M phi) from pre-diseased autoimmune-prone MRL mice (both MRL/+ and MRL/lpr) dramatically underproduce the cytokine interleukin-1 (IL-1) in comparison to M phi from a number of normal strains. In this study we show that IL-1 dysregulation by MRL M phi is fully expressed at birth, and that this defect does not change with time or the development of disease. We also constructed adult irradiation chimeras (consisting of A/J-->MRL and MRL-->A/J mice), and show that M phi isolated from these chimeras display a pattern of IL-1 production indistinguishable from that of the donor strain controls. Moreover, when we constructed a mixed chimera (A/J + MRL-->A/J, the A/J and MRL M phi coexisting within the same animal retained their individual patterns of IL-1 production when isolated by negative selection. Taken together, these results provide the first substantive evidence for an intrinsic defect (IL-1 dysregulation) in M phi from MRL autoimmune-prone mice. PMID- 8223874 TI - Differences in the requirement of antigen-presenting cells for long-term growth among cytomegalovirus-specific Th clonotypes. AB - Functional and molecular studies of in vivo-activated T lymphocytes involved in normal and abnormal immune responses, i.e., cells infiltrating tissues affected by autoimmune processes, require their previous in vitro expansion. Problems such as unavailability of specific antigen(s) (Ag) and/or the requirement of large amounts of autologous peripheral blood mononuclear cells (PBMC) as feeders have prompted the development of alternative expansion methods that circumvent the use of antigen-presenting cells (APC) and/or Ag. We have previously shown that cytomegalovirus (CMV)-specific T cell lines and clones can be efficiently propagated in long-term cultures by stimulation with agonistic monoclonal antibodies (mAb) coated onto polystyrene particles in the absence of APC. However, this and other stimulation protocols may skew the repertoire of clonotypes that proliferate in response to nominal Ag and APC. Here we show that polyclonal populations of CMV-primed and interleukin-2 (IL-2)-stimulated PBMC undergo the same clonotypic selection when induced to grow both by continuous stimulation with CMV and an anti-CD3 mAb presented by APC. This selection, reproduced in three independent expansion experiments, involved the dominant growth of two CMV-specific, IL-2-secreting CD4+ clonotypes sharing J alpha, J beta, V alpha and V beta genes and highly homologous T cell receptor (TcR) junctional sequences. The dominant growth of these 2 clonotypes required a direct T cell/APC interaction since when coated onto polystyrene particles the same mAb induced the selective expansion of another IL-2-secreting CD4+ CMV-specific clonotype expressing a different, yet homologous, TcR heterodimer (used same V alpha and V beta genes), which was underrepresented before expansion (5 vs. 58%). T cell clones belonging to the subdominant clonotype proliferated significantly faster in response to stimulation with anti-CD3 mAb coated onto beads than in response to stimulation with CMV or anti-CD3 mAb presented by APC, possibly due to long-term expansion without APC or antigen. In contrast, neither the dominant clonotypes nor unprimed T cells were able to undergo CD3-mediated expansion in the absence of APC. We conclude that quantitative differences in growth competency among normal Ag-activated T-helper (Th) clonotypes in response to antigenic stimulation can be reproduced by stimulation through the TcR in the absence of TcR occupancy but only in the presence of APC and that certain clonotypes do not require APC for long-term growth in vitro. These data have practical implications for the isolation and repertoire characterization of in vivo-activated T cells from tissues affected by inflammatory, i.e. autoimmune, phenomena. PMID- 8223875 TI - Selective coupling of the T cell antigen receptor to phosphoinositide-derived diacylglycerol production in HPB-ALL T cells correlates with CD45-regulated p59fyn activity. AB - In HPB-ALL T-cells the p59fyn tyrosine kinase is regulated by the CD45 phosphotyrosine phosphatase and plays a critical role in coupling the T cell receptor (TCR) to the generation of intracellular signals which include diacylglycerol (DAG) production and protein kinase C activation. The aim of this study was to determine the phospholipid pools from which the DAG is generated and to identify which phospholipase activities are regulated by the TCR. When CD45+ cells were pre-labeled with [3H]arachidonic acid, CD3-antigen cross-linking stimulated negligible increases in both [3H]DAG and [3H]phosphatidic acid (PA). However, CD3 monoclonal antibody (mAb) induced an increase of 300% in [3H]PA when the cells were permeabilized with streptolysin-O, and this correlated with increased levels of protein tyrosine phosphorylation. Stimulation of [3H]PA production upon CD3 cross-linking was 77% lower in permeabilized CD45- cells than in CD45+ cells, consistent with the reduced activity of p59fyn in CD45- cells. The stimulated production of PA was not mediated by activation of phospholipase D (PLD), although the presence of a G-protein-regulated PLD activity was established. The CD3-induced increase in total inositol phosphates (InsP) in permeabilized cells was similar to the stimulated production of [3H]PA production in both CD45+ and CD45- cells. Dose-response curves for InsP and PA production triggered by CD3 mAb were super-imposable and the production of InsP and PA over a range of Ca2+ concentrations was comparable. Differential labeling of phospholipids with 3H-labeled fatty acids revealed that CD3-induced PA production reflected incorporation of label into the phosphatidylinositol pool. Our data suggest that in HPB-ALL cells the production of DAG following CD3-antigen cross linking can be fully accounted for by the selective coupling of the TCR to breakdown of phosphatidylinositol-(4,5)-bisphosphate as the result of phospholipase C gamma 1 activation. This event correlates with the activity of the CD45-regulated TCR-associated tyrosine kinase, p59fyn. PMID- 8223876 TI - Transcriptional regulation of interleukin-2 gene expression by CD69-generated signals. AB - The 5' flanking region of the human interleukin (IL)-2 gene was investigated for enhancer activity in response to CD69-generated signals, using a chloramphenicol acetyltransferase (CAT)-driven transient expression system in Jurkat cells. The region extending from -317 to +47 relative to the initiation site of IL-2 gene transcription was shown to contain sequences able to respond to CD69 cross linking, by enhancing by about 100% a phorbol 12-myristate 13-acetate (PMA)-plus ionomycin stimulation of CAT activity. A similar increase in CAT activity produced by PMA-plus-anti-CD3 mAb was induced by CD69 cross-linking, while a 200% increase over that obtained by PMA-plus-anti-CD28 mAb stimulation was seen. Analysis of enhancer deletion mutants revealed that proximal AP-1, OCT-1/octamer associated protein and nuclear factor of activated T cells (NFAT) binding regions were all necessary to allow CD69-mediated enhancement of CAT activity. By gel mobility shift analysis, cyclosporin A-sensitive NFAT-binding induction and enhancement of AP-1 binding activity could be detected in nuclear extracts of both Jurkat and peripheral blood T cells after simultaneous CD69 and protein kinase C stimulation. Finally, CD69-mediated signals could increase NFAT and AP-1 binding activity following PMA and ionomycin stimulation in peripheral blood T cells. Collectively, these data suggest that CD69-generated signals participate in the control of the IL-2 gene expression at the transcriptional level, likely acting through NFAT and AP-1 transcription factor complexes. PMID- 8223877 TI - Induction of T cell responses by chimeric bacterial proteins expressing several copies of a viral T cell epitope. AB - A viral T cell epitope was genetically inserted within the periplasmic MalE protein of Escherichia coli in two different permissive insertion sites and resulting hybrid proteins were used to study the in vitro and in vivo immunogenicity of the foreign T cell epitope. Purified hybrid MalE proteins containing the T cell epitope 120-132 (PreS:T) from PreS2 region of hepatitis B virus HBsAg inserted alone or with its adjacent B cell epitope (132-145) were able to induce strong peptide-specific T cell responses in mice. In vitro stimulation of primed lymph node cells or specific T cell hybridomas by the hybrid proteins required processing of the inserted T cell epitope and was inhibited by antigen-presenting cells fixation. The inserted T cell epitope was presented in vitro, in association with appropriate major histocompatibility complex molecules, as efficiently as free synthetic peptide. The in vitro immunogenicity of MalE hybrid proteins was increased by inserting four tandemly repeated copies of PreS:T, either at site 133 or 303. These results were confirmed in vivo by comparing the proliferative responses of lymph node cells from DBA/1 mice primed with MalE hybrid proteins containing one or four copies of PreS:T. Thus, the use of MalE hybrid proteins expressing multiple copies of a given foreign T cell epitope allows the induction of peptide-specific T cell response with a lower dose of priming antigen. PMID- 8223878 TI - Autoimmunity, hyporeactivity to T cell mitogens and lymphoproliferative disorders following neonatal induction of transplantation tolerance in mice. AB - We have reported that, in A/J (A) (H-2a) mice, a partial tolerance to C57BL/10ScSn (B10) (H-2b) skin allografts and a high incidence of lethal lymphoproliferative disorders (LPD) can be induced by the neonatal i.v. injection of 2 x 10(7) semiallogeneic (B10 x A)F1 spleen cells (SC) (Vegh, P., Baranyi, L. and Janossy, T., Cell. Immunol. 1990, 129: 56). In this study, we show that the incidence and mortality of LPD were continuously growing from 1 month of age until the end of the experiment at 1 year (64% and 36%, respectively). Based on histology, 27% of the diseased mice suffered from lymphoid malignancies. In the remaining cases (73%), reactive histopathological changes were seen in the spleen, lymph nodes (LN), liver and kidneys. The proportion of CD4+ T cells in the spleen and LN as well as that of splenic B cells decreased, while the percentages of mature and immature myeloid cells doubled. The total cell number of each (sub)population, however, was elevated in both lymphoid organs. The cells taking part in the lymphoproliferation were of host (A) and not of donor (F1) origin. Preceding the development of apparent LPD, the SC, LN cell and thymus cell suspensions of 1-month-old tolerized mice showed reduced in vitro proliferative responses to T cell and T cell-dependent B cell mitogens (Con A or PWM), while their reactivity to a T cell-independent B cell mitogen (lipopolysaccharide) was essentially unimpaired. This hyporeactivity seems to be functional, because neither histology nor immunophenotyping by flow cytometry revealed significant alterations in the spleen and thymus of such animals, apart from a slight reduction in the ratio of CD4+/CD8+ T cell subpopulations in the spleen. The in vivo T cell-mediated immune response of the tolerized mice was practically normal to third party CBA/Ca (H-2k) allografts. Antithymocyte autoantibodies (ATA) were detected in the sera of 76% of the tolerized mice at 1 month of age (i.e., even before the mass appearance of LPD). ATA as well as antinuclear Ab were present in 65% of the adult tolerized mice, independently of the presence of LPD. Taken together, in A mice neonatally injected with (B10 x A)F1 SC, a partial, specific allograft tolerance and a chronic host-vs.-graft disease-like syndrome developed. The latter is manifested in hyporeactivity to T cell mitogens, development of autoantibodies and, subsequently, in progressive LPD and lymphoid malignancies. PMID- 8223879 TI - Antigens pre-bound to the primate erythrocyte complement receptor via cross linked bispecific monoclonal antibody heteropolymers are rapidly cleared from the circulation. AB - We have used cross-linked, bispecific monoclonal antibody heteropolymers, specific for primate erythrocyte (E) complement receptor (CR1) and target antigen (Ag), to attach 125I-labeled Ag to 51Cr-labeled monkey erythrocytes (E) in vitro. Injection of these sensitized E into monkeys leads to rapid clearance from the circulation of several different 125I-labeled Ag with little, if any sequestration, lysis, or clearance of the E. It should now be possible, in principle, to use the heteropolymer-E system to facilitate the clearance of blood borne pathogens from the circulation. PMID- 8223880 TI - Peptide variants reveal how antibodies recognize major histocompatibility complex class I. AB - The T cell receptor (TcR) on CD8+ T lymphocytes recognizes a complex which consists of a major histocompatibility complex (MHC) heavy chain, beta 2 microglobulin (beta 2M), and peptide on the surface of antigen-presenting cells. Mutational analyses have suggested that the TcR recognizes both the alpha 1 and alpha 2 domains of the heavy chain as well as the peptide. In light of this, it is of interest to know to what extent the heavy chain domains take on distinct conformations when bound to individual peptides. It has recently been shown that antibodies which recognize the Kb MHC complex are sensitive to which peptides are bound in the groove. We have extended this analysis to include eight Kb-specific antibodies, seven of which are peptide sensitive. These antibodies, all of which are allo-antibodies, recognize Kb-bearing cells which, it is now appreciated, have a highly heterogeneous mix of self peptides presented in their grooves. We show that these self peptides also can affect antibody binding. It has been suggested that peptides alter the conformation of the alpha 1 and alpha 2 domains of the heavy chain and that this in turn affects the recognition of Kb by antibody. An alternative hypothesis is that solvent-exposed peptide side chains may prevent the antibody from binding the complex. Using a panel of 128 single amino acid variants of a Kb-binding antigenic peptide from ovalbumin we show that for most Kb-specific antibodies, the second idea is more likely. Those variants which prevent antibody binding are at solvent exposed positions, and in general, the bulkier the side chain, the greater the inhibition of antibody binding. However, in the case of two antibodies, 100.30 and 34.4.20, the peptide residues which affect antibody recognition are buried, suggesting that these antibodies see an alternate conformation of the peptide/MHC complex. PMID- 8223881 TI - Interleukin-4 and interleukin-10 synergize to inhibit cell-mediated immunity in vivo. AB - The lack of cell-mediated (Th1-like) immunity that is often associated with strong humoral immune responses is thought to be due in part to the inhibition of Th1 effector function by the Th2-derived cytokine interleukin-10 (IL-10). This hypothesis, however, is based entirely on results from in vitro studies, wherein IL-10 has been shown to inhibit Th1 cytokine synthesis. In this study we have compared the regulatory effects of both IL-4 and IL-10 on the development of a more complex Th1 effector function in vivo, the development of delayed-type hypersensitivity (DTH) to Leishmania major in mice immune to Leishmania. The results revealed two findings unexpected from in vitro studies with Th1 clones. First, optimal inhibition of the DTH response (up to 70%), assessed by footpad swelling and leukocytic infiltration, required the combination of IL-4 and IL-10, indicating that these two activities synergized to inhibit DTH reactivity. Second, IL-4 inhibited interferon-gamma (IFN-gamma) production by lymph node cells draining the site of antigen challenge as well as did IL-10. The combination of both cytokines was no more effective than either alone. The mechanism by which IL-4 and IL-10 acted to inhibit DTH responses did not appear to be through inhibition of IFN-gamma or tumor necrosis factor production as treatment with antibodies which neutralized these activities failed to inhibit DTH responses. Inhibition of the DTH with IL-4 and IL-10 is the most effective specific regulator of DTH responses reported and the only one capable of modulating tuberculin DTH. These data establish IL-4 and IL-10 as potent inhibitors of Th1 effector function in vivo and suggest their utility in controlling deleterious Th1-mediated inflammatory responses such as occur in some infectious and autoimmune diseases. PMID- 8223882 TI - No independent association between a tumor necrosis factor-alpha promotor region polymorphism and insulin-dependent diabetes mellitus. AB - Several studies have implicated tumor necrosis factor (TNF)-alpha in the pathogenesis of insulin-dependent diabetes mellitus (IDDM). In the present study we analyzed the first reported TNF-alpha gene polymorphism in relation to IDDM. We have made frequence analysis and tested in vitro lipopolysaccharide (LPS) induced TNF-alpha secretion. A significant difference in allele frequency was observed between patients and controls (p = 0.03). However, a very strong association of the uncommon TNF2 allele was observed with the HLA-B8, -DR3 alleles. The relative risk (RR) of TNF2 was 2.2 compared to a RR of 3.1 for DR3. One reason for this difference was the identification of the TNF1 allele on the otherwise strongly IDDM-associated HLA-DR3 haplotype: DQB1*0201, DQA1*0501, DRB1*0301, TNFc2, TNFB*2, TNFa1, TNFb5, B18. Thus, the IDDM-associated TNF2 allele had no DR3-independent value as a disease marker. The LPS-induced TNF alpha production by human monocytes in relation to genotypes demonstrated that TNF1/2 heterozygous individuals had higher, though not statistically significantly (p = 0.08) levels than TNF1-homozygous subjects. However, this difference was rather small, unlikely to be of biological significance and based on the present material we cannot establish the functional importance of this polymorphism. PMID- 8223883 TI - Notice of retraction. PMID- 8223884 TI - Vasodilator effects of quercetin in isolated rat vascular smooth muscle. AB - The effects of quercetin were studied on contractile responses induced by noradrenaline, high KCl, Ca2+ and phorbol 12-myristate,13-acetate in rat aortic strips and on spontaneous mechanical activity in rat portal vein segments. Quercetin, 10(-6)-10(-4) M, inhibited in a concentration-dependent manner the contractions induced by noradrenaline, high KCl and Ca2+, this effect being observed when the drug was added before or after the induced contractions. The spontaneous myogenic portal activity was also inhibited. Mechanical removal of endothelium did not affect the relaxant effects of quercetin on noradrenaline induced contractions. In addition, at the same range of concentrations, quercetin also relaxed the contractions induced by phorbol 12-myristate,13-acetate. Quercetin1 10(-5) and 5 x 10(-5) M, increased the aortic cyclic AMP content. However, pretreatment with 10(-7) M isoprenaline did not modify the relaxant effects of quercetin on noradrenaline-induced contractions and quercetin did not modify the relaxant effects of forskolin, which suggested that the vasodilator effects of quercetin were not mediated by inhibition of cyclic AMP phosphodiesterases. In conclusion, in isolated rat aorta quercetin produced a vasodilator effect that seems to be mainly related to the inhibition of protein kinase C. However, and since this drug exerts multiple biochemical effects, inhibition of other transduction pathways may be involved in this effect. PMID- 8223885 TI - Tachykinins mediate contraction of the human lower esophageal sphincter in vitro via activation of NK2 receptors. AB - The contractile response to natural tachykinins and selective peptide agonists for tachykinin receptors was studied in strips of circular smooth muscle of human lower esophageal sphincter in vitro. The effects of phosphoramidon, which inhibits neutral endopeptidase (EC.3.4.24.11) and of the non-peptide compounds, SR 48968 and CP-96,345, which selectively block NK1 and NK2 receptors, respectively, were also investigated. Substance P, neurokinin A and neurokinin B produced a concentration-dependent contractile response. The rank order of potency was neurokinin A > neurokinin B > substance P. Phosphoramidon (1 microM) potentiated the response to substance P without changing the order of potency of natural tachykinins. The NK2-selective agonist, ([ beta Ala8]neurokinin A-(4 10)), produced a concentration-dependent contraction. The NK1 ([Sar9,Met(O2)11]substance P, 1 microM) and NK3 ([MePhe7]neurokinin B, 1 microM) selective agonists, however, did not exert any contractile effect. The selective NK2 antagonist, SR 48968, potently inhibited in a concentration-dependent (10 nM 1 microM) manner the response to neurokinin A, without affecting the response to carbachol. The selective NK1 antagonist, CP-96,345 (1 microM), did not affect the response to neurokinin A. These results indicate that tachykinins contract the circular muscle of human lower esophageal sphincter, and that this effect is mediated by NK2 receptor stimulation. Moreover, a phosphoramidon-sensitive mechanism plays a role in the regulation of the response to substance P. PMID- 8223886 TI - N(1)-substituted ergolines and tryptamines show species differences for the agonist-labeled 5-HT2 receptor. AB - Previous studies indicated that selected ergolines and tryptamines showed species differences for affinity to the antagonist-labeled 5-HT2 receptor. The present study examined these same compounds for affinity at the agonist-labeled 5-HT2 receptor in rat and squirrel monkey cortical homogenates using [125I]DOI ([125I]1 (2,5-dimethoxy-4-iodophenyl)-2-aminopropane). As seen with the antagonist-labeled 5-HT2 receptor, N(1) alkyl substitution of either the ergolines or tryptamines resulted in a slight increase or no effect on their affinity for the agonist labeled rat 5-HT2 receptor. In contrast, these same N(1) substitutions resulted in significant decreases in affinity for the agonist-labeled monkey 5-HT2 receptor. It was also noted that N(1)-unsubstituted ergolines and tryptamines (such as ergonovine, LY86057, LY193525 and 5-methoxytryptamine) tended to have higher affinity for the monkey versus the rat agonist-labeled receptor. However, the N(1) alkyl-substituted ergolines and tryptamines (such as mesulergine, LY53857, amesergide, N(1)-isopropyltryptamine and N(1)-isopropyl-5 methoxytryptamine) showed significantly lower affinity for the monkey versus the rat 5-HT2 receptor. These data suggest that, at least in relation to the N(1) position, ergolines and tryptamines bind in a similar orientation. These results are also discussed in terms of what amino acid differences between species may account for this structure-activity relationship. PMID- 8223887 TI - Nitric oxide modulates responses to sensory nerve activation of the perfused rat mesentery. AB - The modulatory actions of nitric oxide on sensory nerves were investigated on dilator responses of the perfused rat mesentery to transmural nerve stimulation. N omega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthesis, caused a significant augmentation of vasodilator responses to transmural nerve stimulation, an effect which was abolished by L-arginine. L-NAME had no effect on vasodilator responses to exogenous calcitonin gene-related peptide. In preparations without endothelium L-NAME still caused potentiation of vasodilator responses to transmural nerve stimulation. Methylene blue, an inhibitor of guanylate cyclase, also significantly enhanced vasodilator responses to transmural nerve stimulation. After pretreatment with diethyldithiocarbamate to inhibit superoxide dismutase, vasodilator responses to transmural nerve stimulation were also potentiated. This response was abolished by exogenous superoxide dismutase. These findings suggest that endogenous nitric oxide modulates, in an inhibitory fashion, the actions of sensory nerves in the rat mesentery. The results also suggest that endogenous superoxide dismutase may participate in the regulation of the actions of sensory nerves via control of cellular superoxide anion level. PMID- 8223888 TI - Effects of nor-binaltorphimine on butorphanol dependence. AB - Butorphanol has been shown to act on mu-, delta-, and kappa-opioid receptors. However, the relative involvement of different opioid receptor subtypes in butorphanol dependence is not known. In the present study, nor-binaltorphimine, a long-acting non-peptide kappa-opioid receptor antagonist, was employed to mask central kappa-opioid receptors before and during the induction of butorphanol dependence in rats, so that the involvement of kappa-opioid receptors could be elucidated. The results revealed that treatment with nor-binaltorphimine markedly blocked naloxone-precipitated withdrawal signs of escape behavior, teeth chattering, wet shakes, ptosis, body weight loss, and hypothermia at all doses tested, and attenuated the withdrawal symptoms of forepaw tremors (24 nmol: P < 0.001) and diarrhea (12 nmol: P < 0.05; 24 nmol: P < 0.01). In contrast, nor binaltorphimine had no effect on yawning, ejaculation, nor urination in butorphanol-infused rats undergoing withdrawal. Three days of butorphanol infusion significantly increased KD values (in the cortex and striatum), decreased Bmax (in the cortex only) of [3H]U-69,593 binding, and shifted Ki of nor-binaltorphimine against [3H]U-69,593 (4.5 nM) binding in the cortex by more than 10-fold. Treatment with nor-binaltorphimine blocked the effects of butorphanol on kappa-opioid receptors. It is therefore concluded that kappa opioid receptors are involved in mediating escape behavior, teeth-chattering, wet shakes, forepaw tremors, ptosis, diarrhea, weight loss, and hypothermia in butorphanol-dependent rats undergoing withdrawal. Furthermore, kappa-opioid receptors become desensitized to agonists (in the cortex and striatum), down regulated (in the cortex), and supersensitive to antagonists in butorphanol dependent rats. PMID- 8223889 TI - Effects of three K+ channel openers on airways and pulmonary circulation in the isolated guinea-pig lung. AB - The antispasmodic and spasmolytic effects of levcromakalim (BRL 38227), aprikalim (RP 52891) and pinacidil were investigated in airways and pulmonary vessels of the isolated guinea-pig perfused lung. In airways, the three drugs exhibited modest antispasmodic properties, and pinacidil was more potent than levcromakalim and aprikalim against the contractions induced by carbachol (0.001-10 microM) or K+ (5-50 mM). Whereas levcromakalim and aprikalim acted only at low concentrations of K+, the rightward shift of the K+ concentration-effect curve produced by pinacidil was observed at all K+ concentrations (5-50 mM), suggesting that a mechanism of action other than K+ channel opening is involved in the effects of pinacidil. Pinacidil (0.3-100 microM) had the greatest spasmolytic effect in airways precontracted by carbachol (0.3 microM). The three K+ channel openers were equipotent against the sustained contractions of airways induced by 25 or 30 mM K+ and their spasmolytic activity was more marked against contractions induced by low rather low than high K+ concentrations. Levcromakalim and aprikalim were more effective as relaxant than as antispasmodic drugs. In K(+)-precontracted pulmonary vessels, the relaxant activity of pinacidil and levcromakalim was more pronounced than that observed in airways, suggesting a vascular selectivity of these drugs. PMID- 8223890 TI - Effects of CS-518, a thromboxane synthase inhibitor, on eosinophil function. AB - The effects of CS-518 (sodium 2-(1-imidazolylmethyl)-4,5-dihydrobenzo[b]thiophene 6-carboxylate) , a thromboxane A2 synthase inhibitor, on eosinophil accumulation and activation were investigated in an experimental asthmatic guinea pig model and several cellular models. In the in vivo studies, CS-518 inhibited the biphasic eosinophil accumulation in the bronchoalveolar lavage fluid, potently in the early phase, but less potently in the delayed phase. On the other hand, even at the lower dose, CS-518 completely inhibited the hypodensity of eosinophils in the delayed phase. In the in vitro studies, CS-518 suppressed thromboxane A2 production and potentiated prostaglandin I2 production from guinea pig eosinophils. Moreover, CS-518 and prostaglandin I2 suppressed chemotaxis, peroxidase release and superoxide generation in guinea pig eosinophils. In addition, the present studies provide further support for the possibility that thromboxane A2 and prostaglandin I2, which are produced in bronchoalveolar tissue and within eosinophils, are involved in modulation of eosinophil function and suggest that CS-518 is a potent inhibitor of eosinophil activation. PMID- 8223891 TI - Effect of a Ca2+ entry blocker, nilvadipine, on hearing disturbances and equilibrium dysfunction caused by microcirculatory disorders of the rat inner ear. AB - We evaluated the effects of Ca2+ entry blockers, nilvadipine and flunarizine, on microcirculatory disorders of the inner ear and on blood flow in the inner ear of rats. Under sodium pentobarbital anesthesia, the middle ear was opened by a ventrolateral approach. A green light (wave length 540 nm) was applied to the cochlea or the vestibule to induce a hearing disturbance or equilibrium dysfunction as a result of inner ear microcirculatory disorders, while rose bengal solution was infused intravenously. In a hearing disturbance model, a compound cochlear nerve action potential was recorded by electrocochleography every minute after the beginning of illumination. The sound stimulus was an 8 kHz sine wave 100 dB normal hearing level. The action potential was calculated 128 times. The action potential disappeared about 12 min after the beginning of illumination. In another model of equilibrium dysfunction, the photoillumination was applied for 40 min under the infusion of rose bengal. The behavior of rats was observed in the swimming test and nystagmus was recorded 24 h after the completion of photoillumination. In a separate experiment, blood flow in the inner ear was measured with a laser Doppler flowmeter under sodium pentobarbital anesthesia. In this study, both nilvadipine and flunarizine prolonged the time required for complete suppression of the action potential, prevented equilibrium dysfunction in the swimming test and reduced the occurrence of nystagmus. Flunarizine significantly increased inner ear blood flow and nilvadipine failed to decrease blood flow in the inner ear, despite a reduced systemic blood pressure. In conclusion, Ca2+ entry blockers may prevent microcirculatory disorders of the inner ear in rats. PMID- 8223892 TI - Restraint stress potentiates analgesia induced by 5'-N-ethylcarboxamidoadenosine: comparison with morphine. AB - mu-Opioid receptor agonists, e.g. morphine, produce analgesia that can be potentiated by restraint stress. Adenosine receptor agonists, e.g. 5'-N ethylcarboxamidoadenosine (NECA), also produce analgesia. To determine if adenosine-induced analgesia is also potentiated by stress, dose- and time-effect curves for NECA (0.01-0.1 mg/kg s.c.) were generated in adult male Sprague-Dawley rats either unrestrained or restrained in Plexiglas cylinders, using the tail flick assay. Morphine (1.0-5.6 mg/kg s.c.) was also tested for comparison. Both compounds produced dose-dependent increases in tail-flick latencies. This effect of both drugs was potentiated in restrained rats. The opioid receptor antagonist, naltrexone (1.0 mg/kg s.c.) blocked completely the effect of morphine in both groups and attenuated the stress-induced potentiation of NECA-induced analgesia. The adenosine receptor antagonist, caffeine (10.0 mg/kg s.c.), blocked the analgesic effect of NECA but not that of morphine. These results indicate that adenosine-mediated analgesia is potentiated by restraint stress and suggest a role for endogenous opioids in the mediation of stress-induced potentiation of analgesia. PMID- 8223893 TI - The thyroadrenal axis, food deprivation and retention of a behavioural response. AB - The effects of adrenalectomy on retention of the immobile response in the Porsolt swimming rat test are equally reversed by dexamethasone, ketocyclazocine and thyroxine; the effects of hypothyroidism are similarly equally well reversed by all three agents, or by adrenalectomy. Animals food deprived for 24 h lose the response, but retain it after 48 h food deprivation; the latter effect is blocked by adrenalectomy. These counterintuitive findings suggest hitherto unrecognized commonalities and/or interactions in the metabolic effects of the thyroid and adrenal glands. PMID- 8223894 TI - Neurochemical profile of the selective and silent 5-HT1A receptor antagonist WAY100135: an in vivo microdialysis study. AB - The neurochemical profile of the selective 5-HT1A receptor antagonist WAY100135 [N-tert-butyl 3-4-(2-methoxyphenyl) piperazin-1-yl-2-phenylpropanamide dihydrochloride] and its enantiomers at the somatodendritic 5-HT1A receptor was determined by studying the effects of these compounds on 5-HT (5 hydroxytryptamine, serotonin) release in the rat hippocampus using in vivo microdialysis. (+/-)-WAY100135, (+)-WAY100135 and (-)-WAY100135 (all at 10 mg/kg s.c.) had no significant effect on extracellular levels of 5-HT in the hippocampus demonstrating that these compounds are devoid of 5-HT1A receptor agonist properties. In contrast, the 5-HT1A receptor agonist 8-hydroxy-2-(di-n propylamino) tetralin (8-OH-DPAT) (0.1 mg/kg s.c.) and the partial agonists BMY 7378 (1.0 mg/kg s.c.) and buspirone (5 mg/kg s.c.) significantly decreased hippocampal 5-HT. Pretreatment with (+/-)-WAY100135 (at 10 mg/kg s.c.) and (+) WAY100135 (at 1.0-10 mg/kg s.c.) completely blocked the 8-OH-DPAT-induced decrease in 5-HT release demonstrating that these compounds are antagonists at the somatodendritic 5-HT1A autoreceptor. (-)-WAY100135 at a dose of 10 mg/kg s.c. had no significant effect on the 8-OH-DPAT-induced inhibition of 5-HT release. (+/-)-WAY100135 had no significant effect on extracellular levels of dopamine in the rat hippocampus but significantly increased extracellular levels of noradrenaline. The mechanism underlying the increase in noradrenaline is unknown at present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223895 TI - The effect of 5-HT receptor ligands on the uptake of [3H]5-hydroxytryptamine into rat cortical synaptosomes. AB - The effect of 5-HT receptor agonists and antagonists to inhibit [3H]5-HT uptake was investigated in rat cortical synaptosomes. The 5-HT (5-hydroxytryptamine) uptake inhibitors paroxetine and fluoxetine yielded pKi values of 8.41 +/- 0.12 and 7.43 +/- 0.06 respectively. The 5-HT3/5-HT4 receptor antagonist tropisetron and the 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) had similar inhibitory potencies to cocaine (pKi values of 6.58 +/- 0.04, 6.47 +/- 0.14 and 6.45 +/- 0.12 respectively). The dopamine and noradrenaline uptake inhibitors GBR12909 and desipramine had comparable values of 6.5 +/- 0.05 and 6.13 +/- 0.07. Other 5-HT receptor ligands had pKi values less than 6.0 (R(+) zacopride, MDL72222, R(+)/S(-)-zacopride) or 5.0 (5-methoxytryptamine, m chlorophenylbiguanide, S(-)-zacopride, SDZ205-557, ondansetron and renzapride). It is concluded, with the possible exception of tropisetron and 8-OH-DPAT, that it is unlikely that the effects of the 5-HT receptor ligands to inhibit 5-HT uptake contribute to their effects in vivo. PMID- 8223896 TI - Different mode of action of endothelin-1 in parietal and visceral veins. AB - The mode of contractile effect of endothelin-1 in venous smooth muscles was evaluated using three isolated veins, the jugular vein, the portal vein and the inferior caval vein of dogs. The contractile responses to endothelin-1 were attenuated by atropine and augmented by neostigmine in the inferior caval and portal veins, but were not affected by either agent in the jugular vein. We conclude that the contractile responses of visceral veins to endothelin-1 are partly mediated via endogenously released acetylcholine. PMID- 8223897 TI - Arenobufagin, a compound in toad venom, blocks Na(+)-K+ pump current in cardiac myocytes. AB - The effects of arenobufagin, a bufadienolide isolated from toad venom, on the Na+/K+ pump were investigated by recording the whole-cell current of dissociated cardiac myocytes. Arenobufagin blocked the Na+/K+ pump current in a dose dependent manner with a half-maximal concentration of 0.29 microM and a Hill coefficient of 1.1. Arenobufagin is 3.5-fold more potent than ouabain. PMID- 8223898 TI - Comparative action of new highly potent bradykinin receptor antagonists in the guinea-pig trachea. AB - We investigated the effect of two new bradykinin receptor antagonists, D Arg0[Hyp3,D-HypE(trans-propyl)7,Oic8]bradykinin (NPC 17731) and D-Arg0[Hyp3,D HypE(trans-thiophenyl)7,Oic8]bradykinin (NPC 17761) on [3H]bradykinin binding and on bradykinin-induced contraction of the guinea-pig trachea. Both of these compounds inhibited [3H]bradykinin binding with an affinity similar to that of unlabelled bradykinin. NPC 17731 inhibited bradykinin-induced contraction in a non-competitive manner, whereas NPC 17761 showed competitive antagonism. Therefore, NPC 17761 could contribute to the investigation of the bradykinin receptors in guinea-pig airways. PMID- 8223899 TI - Nitric oxide, but not interleukin-1, mediates the local blood flow response to lipopolysaccharide in rabbit skin. AB - Lipopolysaccharide can mimic many aspects of the inflammatory response to gram negative bacteria. In rabbit and rat skin, lipopolysaccharide induces neutrophil accumulation and an increase in blood flow. Interleukin-1, tumor necrosis factor alpha and the vasodilator, nitric oxide (NO) have been implicated in the biological responses to lipopolysaccharide and gram-negative bacteria. In the present study, we characterized the local vascular response to E. coli lipopolysaccharide in rabbit skin and investigated its dependence on de novo protein synthesis, NO, interleukin-1 and neutrophils. The local vascular response to the intradermal injection of lipopolysaccharide was determined by measuring changes in local blood flow with a laser-Doppler flowmeter. Injections of lipopolysaccharide (3, 10 and 30 micrograms/site) induced a dose-related increase in blood flow, with a maximum at 2 h. The response to 10 micrograms/site lipopolysaccharide was abolished by co-injection of actinomycin D (4 x 10(-9) mol/site), reduced by 60% by NG-nitro-L-arginine methyl ester (10(-7) mol/site) and suppressed by 82% in rabbits pretreated with dexamethasone (2 mg/kg, i.v.). However, the lipopolysaccharide-induced increase in blood flow was not affected by co-injection of recombinant human interleukin-1 receptor antagonist (7.0 x 10( 11) mol/site) or by systemic neutrophil depletion. These results demonstrate that lipopolysaccharide causes delayed vasodilatation of the rabbit cutaneous microvasculature which depends on protein synthesis and involves in part the release of NO. However, this response does not depend on interleukin-1 or neutrophils. The vasodilator effect of lipopolysaccharide may involve the expression of the inducible form of NO synthase in the vessel wall, through a mechanism independent of interleukin-1. PMID- 8223900 TI - Increased plasma levels of endothelin during anaphylactic shock in the guinea pig. AB - Intravenous injection of ovalbumin into actively and passively sensitized guinea pig resulted in acute circulatory collapse. The plasma level of immunoreactive endothelin rose from 22 +/- 2 to 40 +/- 7 fmol/ml (n = 12, P < 0.01) and 29 +/- 5 fmol/ml (n = 12, P < 0.01) in actively and passively sensitized animals, respectively, within 5 min of antigen challenge, and it remained significantly higher in actively sensitized animals that survived for 15 min. The plasma immunoreactive endothelin level was inversely correlated with arterial blood pO2, but not with pH or pCO2, both in actively (rs = -0.585, n = 20, P < 0.05) and passively sensitized animals (rs = -0.558, n = 20, P < 0.05). When non-sensitized animals were bled (5 and 20 ml/kg body weight), the plasma immunoreactive endothelin level remained unchanged. These results suggest that the elevated plasma level of immunoreactive endothelin during anaphylactic shock is independent of hypotension, hypovolemia and respiratory insufficiency. PMID- 8223901 TI - Conventionalization of germ-free rats reverses the disability of rhein anthrone to induce laxation. AB - This study shows that rhein anthrone has no laxative potency in germ-free rats because after intracaecal administration of a dose of 50 mg/kg the large intestine transit exceeded 240 min. The time course of the laxative potency of rhein anthrone injected intracaecally was evaluated after peroral inoculation of germ-free rats with the caecal contents of conventional rats. Large intestine transit was measured at consecutive periods, on days 0, 1, 2, 3 and 5 after peroral inoculation. It appeared that 1 day after peroral inoculation the laxative potency of rhein anthrone was already established (large intestinal transit < 10 min) and laxation remained on the following days (days 2, 3 and 5). We concluded that rhein anthrone is inactive in germ-free rats and acquires laxative potency after peroral inoculation of germ-free rats with caecal contents of conventional rats. PMID- 8223902 TI - The glycine/NMDA receptor ligand (+)-HA-966 but not D-cycloserine has potent antidystonic efficacy in a genetic animal model of dystonia. AB - The effects of R-(+)-HA-966 ((+)-3-amino-1-hydroxypyrrolid-2-one), a low-efficacy partial agonist of the glycine modulatory site of the NMDA receptor complex, were studied in an inbred line of Syrian golden hamsters with generalized dystonia, a frequent movement disorder in humans. The effects of R-(+)-HA-966 were compared with those of D-cycloserine, a glycine/NMDA receptor ligand with higher intrinsic activity. R-(+)-HA-966, 30-60 mg/kg i.p., potently reduced the severity of dystonic attacks in the mutant hamster model of dystonia without inducing any behavioural adverse effects. D-Cycloserine did not exert antidystonic activity at i.p. doses of 10-40 mg/kg, which might be due to its much higher intrinsic activity at the glycine site. The data indicate that the antidystonic effect of (+)-HA-966 is related to antagonism of NMDA receptor-mediated excitatory neurotransmission. PMID- 8223903 TI - Antidepressant-like effect of LY228729 as measured in the rodent forced swim paradigm. AB - The novel 5-HT1A receptor agonist, LY228729, was tested to see if it would produce antidepressant-like behavioural effects in the rodent forced swim model. The tricyclic antidepressant, imipramine, as well as the 5-HT1A receptor agonists, 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and gepirone, produced dose-related decreases in immobility in the forced swim test following subchronic treatment in rats. LY228729, when given over a three injection course, as with the other compounds, significantly reduced immobility at doses of 1 and 3 mg/kg. Subsequent analysis of locomotor activity revealed no increases in behavior, so that general changes in activity could not account for the reduction in immobility time in the forced swim. These results suggest that LY228729 may have clinical antidepressant efficacy. PMID- 8223904 TI - Gliquidone, an ATP-dependent K+ channel antagonist, antagonizes morphine-induced hypermotility. AB - The effect of gliquidone, an ATP-dependent K+ (KATP) channel blocker, on morphine induced hypermotility in mice was studied. Morphine (5-40 mg/kg s.c.) dose dependently increased ambulatory activity. Gliquidone (10 micrograms/mouse i.c.v.) induced a parallel displacement to the right of the morphine dose response curve. Moreover, gliquidone (10 and 40 micrograms/mouse i.c.v.) produced a dose-dependent antagonism of morphine (20 mg/kg s.c.)-induced hypermotility. These results suggest that KATP channels are involved in morphine-induced hypermotility. The present data, together with those of previous studies showing antagonism by KATP channel blockers of morphine-induced antinociception and hyperthermia, further indicate that the opening of KATP channels plays an important role in the mechanism of action of morphine. PMID- 8223905 TI - Inhibition of neurogenic plasma exudation and bronchoconstriction by a K+ channel activator, BRL 38227, in guinea pig airways in vivo. AB - Intravenous administration of a K+ channel activator, BRL 38227, inhibited cigarette smoke-induced plasma exudation in guinea pig airways in vivo in a dose dependent manner with an approximate ED50 of 6 microgram/kg. BRL 38227 also inhibited vagally induced plasma exudation and bronchoconstriction but did not inhibit substance P-induced plasma exudation or neurokinin A-induced bronchoconstriction. K+ channels modulate neurotransmission in the airways and K+ channel activators may have therapeutic potential in bronchial diseases including asthma and chronic bronchitis. PMID- 8223906 TI - Modulation of acetylcholine release from the myenteric plexus of guinea-pig ileum by 5-lipoxygenase metabolites of arachidonic acid. AB - Eicosatetraenoic acid, an inhibitor of 5- and 12-lipoxygenase, and AA861, a selective inhibitor of 5-lipoxygenase, dose dependently inhibited acetylcholine release from the myenteric plexus of guinea-pig ileum induced by electrical field stimulation. Metabolites of arachidonic acid produced by the 5-lipoxygenase pathway, such as 5-hydroxyeicosatetraenoic acid (5-HETE), leukotriene C4, D4 and E4, reversed the inhibitory effect of AA861. Among them, leukotriene D4 was the most potent, having an EC50 value of about 3 nM. The present study shows for the first time that 5-lipoxygenase metabolites may have a modulatory effect on acetylcholine release in the myenteric plexus of guinea-pig ileum. PMID- 8223907 TI - Metabotropic glutamate receptors contribute to the induction of long-term depression in the CA1 region of the hippocampus. AB - Long-term depression of synaptic transmission was induced following the prior induction of long-term potentiation in the CA1 region of rat hippocampal slices. We show that the induction of this form of synaptic depression can be prevented by (+)-alpha-methyl-4-carboxyphenylglycine, a selective antagonist of metabotropic glutamate receptors. PMID- 8223908 TI - Heterogeneity of endothelin/sarafotoxin receptors mediating contraction of the human isolated saphenous vein. AB - We investigated the effect of the ETA receptor antagonist, BQ-123 (0.1 and 1 microM), on contraction of the human isolated saphenous vein induced by endothelin-1 or sarafotoxin S6b. Contraction in response to endothelin-1 was not affected by BQ-123. In contrast, BQ-123 biphasically attenuated the contractions due to sarafotoxin S6b. These data indicate that (i) endothelin-1 induces contractions of the human saphenous vein via a non-ETA receptor and (ii) contractions in response to sarafotoxin S6b are mediated in part via a receptor different from the receptor mediating contraction due to endothelin-1. PMID- 8223909 TI - Isomeric selectivity at dopamine D3 receptors. AB - Racemic 7-hydroxy-N,N-dipropylaminotetralin (7-OH-DPAT) shows greater affinity for limbic-selective dopamine D3 receptors than for more ubiquitous dopamine D2 receptors. R(+)-7-OH-DPAT was prepared and evaluated in radioreceptor assays using membranes of fibroblasts expressing the human dopamine D3 receptor as well as rat striatal membranes containing dopamine D2 receptors. This enantiomer had 2 fold greater D3 affinity than the racemate and similarly greater D3 vs. D2 selectivity (64-fold). The results may facilitate development of D3 selective agents and evaluation of functions of these receptors. PMID- 8223910 TI - Hexahydrobenzo[a]phenanthridines: novel dopamine D3 receptor ligands. AB - We report that certain substituted hexahydrobenzo[a]phenanthridines are novel high affinity ligands selective for the dopamine D3 receptor. These data demonstrate that substitutions on the heterocyclic nitrogen and the pendant phenyl ring of this nucleus cause a marked increase in both affinity and selectivity for dopamine D3 vs. D2 receptors. Thus, these compounds represent important new tools to study the pharmacology of dopamine D3 receptors, and may also provide an opportunity for the synthesis of new radioligands for dopamine D3 receptors. PMID- 8223911 TI - Serotonin activates Cl- channels in the apical membrane of rat choroid plexus epithelial cells. AB - The effects of serotonin on ion channel activity in epithelial cells from rat choroid plexus were examined. Serotonin (50 nM, 500 nM and 1 microM) stimulated channel activity in cell-attached patches. The current-voltage (I-V) relationship for the serotonin-activated channel gave a conductance of 26.6 +/- 1.5 pS and the current reversed at an applied electrode potential (-Vp) = 15.3 +/- 3.3 mV with a KCl-rich electrode solution (n = 8). Similar I-V relationships were obtained using electrode solutions in which K+ was replaced by other cations (Na+ and N methyl-D-glucamine), suggesting that the serotonin-activated channels are selective to Cl-. The effect of 1 microM serotonin on channel activity was inhibited by ritanserin (30 and 100 nM) which has a high affinity for serotonin 5 HT1C receptors and 5-HT2 receptors. Spiperone (30 nM), which binds weakly to 5 HT1C receptors but has a high affinity for 5-HT2 receptors, did not inhibit the actions of serotonin. These data suggest that serotonin increases Cl- channel activity by acting on the 5-HT1C receptors on the epithelium. PMID- 8223912 TI - Modulation of rat brain endogenous dopamine metabolism by new inhibitors of catechol O-methyltransferase. AB - The extraneuronal and intraneuronal metabolism of rat brain endogenous dopamine was stimulated by amphetamine (5 mg/kg) and pimozide (2 mg/kg), respectively. Additional metabolic effects of two inhibitors of catechol O-methyltransferase (entacapone and tolcapone (both 30 mg/kg)) and a putative central uptake2 inhibitor (CGP 28014 (30 mg/kg)) were assessed. Amphetamine increased striatal dopamine and 3-methoxytyramine and decreased 3,4-dihydroxyphenylacetic acid (DOPAC) levels. The latter two effects were reversed by tolcapone and CGP 28014, but not by entacapone. Tolcapone, CGP 28014 and even entacapone decreased striatal homovanillic acid (HVA) levels. Pimozide-induced striatal DOPAC levels were further increased by tolcapone and CGP 28014. Both substances also decreased striatal HVA levels. Striatal 3-methoxytyramine levels were significantly lowered only by tolcapone. Our results show that enhanced central dopamine metabolism is modified by the inhibition of catechol O-methyltransferase even in the absence of L-3,4-dihydroxyphenylalanine (L-DOPA). The results also suggest that the mechanism of action of CGP 28014 may be other than true inhibition of catechol O methyltransferase. PMID- 8223913 TI - Further evidence for a functional dorsal-ventral division of the rat striatum: GABAergic involvement in oral movements. AB - Interactions between the gamma-aminobutyric acid (GABA)ergic system and the dopaminergic and cholinergic systems in the control of jaw movements, measured with a phototransducer system, were investigated in both dorsal and ventral regions of the rat striatum. Muscimol (25 and 50 ng/0.2 microliters) injected into the dorsal striatum did not affect jaw movements induced by apomorphine (0.2 mg/kg i.v.) or pilocarpine (4 mg/kg i.v.), but when injected into the ventral striatum it inhibited these jaw movements and those induced by carbachol (1 micrograms/0.2 microliters) injected into the ventral striatum. Picrotoxin (250 and 500 ng/0.2 microliters) injected into the dorsal striatum enhanced the effects of apomorphine (0.2 mg/kg) but not those of pilocarpine. When injected into the ventral striatum, picrotoxin did not affect apomorphine-induced oral movements but enhanced the effects of pilocarpine and carbachol. The benzodiazepine, flunitrazepam (100 ng/0.2 microliters), had no effect when injected in the dorsal striatum, and showed some inhibitory effects on dopaminergic and cholinergic oral movements when injected in the ventral striatum. The results suggest that the striatal GABAergic inhibitory effect on dopaminergic and cholinergic function is regionally specific, supporting a dorsal/ventral functional division of the rat striatum. The results also suggest that oral movements induced by dopaminergic and cholinergic drugs are distinct forms of repetitive oral behaviour. PMID- 8223914 TI - Effect of gangliosides on diacylglycerol content and molecular species in nerve from diabetic rats. AB - The effects of ganglioside treatment on 1,2-diacylglycerol content and on molecular species in 1,2-diacylglycerol, phosphatidic acid and total diacylglycerolipids, as well as Na+,K(+)-ATPase activity, were examined in sciatic nerves from streptozotocin-induced diabetic rats. Beginning 2 weeks after induction of diabetes, animals were administered mixed bovine brain gangliosides, AGF1, an inner ester derivative of this mixture, or saline for 5 weeks. The levels of 1,2-diacylglycerol and arachidonoyl-containing molecular species in age matched non-diabetic animals were not affected by ganglioside treatment. In nerves from saline-treated diabetic animals, 1,2-diacylglycerol levels were not reduced, but both Na+,K(+)-ATPase activity and all arachidonyl-containing species except for 18:0/20:4 1,2-diacylglycerol were significantly decreased. The content of 1,2-diacylglycerol was lowered by 23 and 16% in bovine brain ganglioside and AGF1-treated diabetic animals, respectively, and the quantity of 18:0/20:4 1,2 diacylglycerol was also selectively reduced. Ganglioside administration did not affect the diminished levels of arachidonoyl-containing molecular species in 1,2 diacylglycerol, phosphatidic acid or diacylglycerolipids in nerve from diabetic rats. In the same nerves, bovine brain gangliosides partially and AGF1 completely restored Na+,K(+)-ATPase activity. The results suggest that gangliosides depress the content of total 1,2-diacylglycerol and the quantity of 18:0/20:4 1,2 diacylglycerol, specifically, in diabetic nerve. The possible relationship between the corrective action of gangliosides on Na+,K(+)-ATPase activity and the effect of these substances on 1,2-diacylglycerol molecular species composition and metabolism is discussed. PMID- 8223915 TI - Induction of bradykinin B1 receptor-mediated relaxation in the isolated rabbit carotid artery. AB - Responses to bradykinin and to the bradykinin B1 receptor agonist des-Arg9 bradykinin were studied in freshly isolated rabbit carotid artery rings and in rings after a 5-h period of incubation. In freshly isolated rings precontracted with noradrenaline, neither bradykinin nor des-Arg9-bradykinin changed the tension whereas acetylcholine relaxed the vessel in a concentration-dependent manner. After incubation, des-Arg9-bradykinin, and to a lesser extent bradykinin, produced an endothelium-dependent relaxation. This response was abolished by endothelium removal, N omega-nitro-L-arginine or cycloheximide but was unaffected by indomethacin. In contrast, the response to acetylcholine was unaffected by cycloheximide and was partially inhibited by N omega-nitro-L-arginine. In addition, the relaxation curve for des-Arg9-bradykinin was markedly shifted (44 fold) by the selective bradykinin B1 receptor antagonist, des-Arg9 [Leu8]bradykinin (3 microM) and was unaffected by the bradykinin B2 receptor antagonist, Hoe 140 (1 microM). We conclude that in vitro incubation of the rabbit carotid artery induced endothelial bradykinin B1 receptors coupled to the release of endothelium-derived nitric oxide. PMID- 8223916 TI - Effects of dotarizine on 45Ca2+ movements and contractile responses in vascular smooth muscle. AB - The inhibitory effects of dotarizine on 45Ca2+ movements and contractile responses were studied and compared, using the same parameters measured in rabbit aorta and basilar smooth muscle. Dotarizine 10(-9)-10(-5) M inhibited the contractile responses induced by high K+ (80 mM), noradrenaline (10(-6) M) or 5 hydroxytryptamine (5-HT, 10(-5) M). These effects were observed when dotarizine was added before or after the induced contractions and were more potent in basilar arteries than in aorta. Moreover, dotarizine at concentrations less than 10(-6) M did not modify the contractile response obtained in aorta rings. Contractile responses induced by the addition of Ca2+ to Ca(2+)-free high-K+ solution were also concentration dependently inhibited by dotarizine 10(-7)-10( 6) M in aorta and basilar arteries. Dotarizine also inhibited the contractile response induced by caffeine (20 mM) in aortic rings incubated in normal or in Ca(2+)-free medium. Dotarizine reduced the 45Ca(2+) uptake stimulated by high K+, noradrenaline or 5-HT even in the aorta or basilar artery, but the inhibition was greater in basilar arteries than in aorta. These results suggest that, in rabbit, dotarizine inhibits Ca2+ entry through Ca2+ channels, being more selective for the basilar artery, probably by acting on multiple sites to decrease the availability of intracellular free Ca2+ required for activation. PMID- 8223917 TI - Common and uncommon behavioural effects of antagonists for different modulatory sites in the NMDA receptor/channel complex. AB - The behavioural significance of the N-methyl-D-aspartate (NMDA) receptor/channel complex was investigated in rats, using different types of antagonists including D-2-amino-phosphonovaleric acid, (+/-)-3-(2-carboxypiperazin-4-yl)-propyl-1 phosphonic acid, 2-amino-4,5-(1,2-cyclohexyl)-7-phosphonoheptanoic acid, 7-Cl kynurenate, ifenprodil, phencyclidine (PCP), MK-801, ketamine, (+/-)-N allylnormetazocine, dextromethorphan, ZnCl2, and MgCl2. All antagonists produced an impairment of learning and muscle relaxation when administered i.c.v. Frequent circling was characteristically produced by ligands for PCP binding sites and was unaffected by concomitant administration of a competitive NMDA receptor antagonist. These results suggest that the NMDA receptor/channel plays an important role in regulating learning/memory processes and muscle tone, whereas PCP binding sites unassociated with the NMDA channel are involved in the production of characteristic circling behaviour. PMID- 8223918 TI - Lack of cross-tolerance between U69,593 and bremazocine suggests kappa-opioid receptor multiplicity in mice. AB - The development of tolerance, and the possibility of cross-tolerance, to the kappa-opioid receptor-mediated antinociceptive effects of U69,593 and bremazocine was studied in mice. U69,593 and bremazocine elicited dose-related and kappa receptor-mediated antinociception following i.c.v. administration to mice. After a 3 day treatment regimen (twice daily injections) with an approximate antinociceptive A90 dose (90 nmol, i.c.v.) of U69,593, tolerance developed as demonstrated by a 5.6-fold rightward shift of the U69,593 dose-response line. The i.c.v. dose-response line for bremazocine was unaltered in U69,593-tolerant mice. Pretreatment with an approximate antinociceptive A90 dose of bremazocine (30 nmol, i.c.v.) for 3 days also produced tolerance as shown by a greater than 15 fold rightward shift in the bremazocine antinociceptive dose-response line. The i.c.v. dose-response line for U69,593 was unaltered in bremazocine-tolerant mice. These data demonstrate that while tolerance develops to the antinociceptive effects of both U69,593 and bremazocine, a two-way lack of cross-tolerance can be demonstrated between these kappa-agonists in this endpoint. These data suggest mechanistic differences in the antinociceptive effects of these kappa-agonists. Such suggestions are consistent with antinociceptive action of these agonists at subtypes of kappa-receptors. PMID- 8223919 TI - Analysis of the 5-HT1C receptor and the serotonin uptake site in fawn-hooded rat brain. AB - Both the 5-HT1C receptor and the 5-HT uptake binding sites were measured in Fawn Hooded, Sprague-Dawley and Wistar rats. Five brain regions were examined: frontal cortex, hippocampus, striatum, hypothalamus, and brainstem. We found significant differences in the Bmax and Kd values in various brain regions comparing Fawn Hooded rats, with Sprague-Dawley and Wistar animals. The regional differences in receptor number and affinity in both the 5-HT1C receptor and the 5-HT uptake site in the Fawn-Hooded strain, relative to Wistar and Sprague-Dawley animals, provide support for the use of the Fawn-Hooded rat in serotonin dysfunction studies. PMID- 8223920 TI - Pharmacological characteristics of tachykinin receptors mediating acetylcholine release from neonatal rat spinal cord. AB - The pharmacological profiles of tachykinin receptors mediating the release of acetylcholine were examined in the isolated spinal cord of the neonatal rat. The acetylcholine release evoked by neurokinin A or acetyl-[Arg6,Pro9]substance P-(6 11) was depressed by the tachykinin antagonists, spantide and GR71251 at 10 microM, whereas the release evoked by substance P (0.3 microM) or neurokinin B (0.3 microM) was not affected by these tachykinin antagonists. Polymerase chain reaction amplification of rat spinal cord cDNA and sequence analysis revealed the presence of a substantial amount of fragments having sequences identical to that of the NK1 or NK3 receptor, but only a few having a sequence identical to that of the NK2 receptor. These results suggest that in the neonatal rat spinal cord a novel subtype of tachykinin receptor similar but not identical to the classical NK1 receptor is involved in tachykinin-evoked acetylcholine release. PMID- 8223921 TI - Cardiovascular effects produced by systemic injections of nitro blue tetrazolium in the rat. AB - Nitro blue tetrazolium is a powerful electron acceptor which is widely used to localize NADPH-dependent flavin-containing enzymes known as NADPH diaphorases. By accepting electrons, nitro blue tetrazolium is known to inhibit the activity of these enzymes. The present study examined the effects of intravenous nitro blue tetrazolium on arterial blood pressure and regional blood flows in urethane anesthetized rats. Nitro blue tetrazolium (0.1-5 mol/kg) produced a dose dependent hypotension and differential effects on regional hemodynamics including decreases in hindquarter and mesenteric vascular resistances and a marked increase in renal resistance. These results demonstrate that systemic administration of nitro blue tetrazolium produces profound hemodynamic effects, the mechanisms of which remain to be elucidated. PMID- 8223922 TI - The regulatory effect of bradykinin on the actions of sensory nerves in the perfused rat mesentery is mediated by nitric oxide. AB - In the perfused rat mesentery, when adrenergic nerves were blocked with guanethidine and vascular smooth muscle tone was increased with methoxamine, transmural field stimulation caused a dilator response. Bradykinin significantly suppressed vasodilator responses to a transmural field stimulation in a concentration-dependent manner. After pretreatment with saponin to damage endothelial cells, bradykinin still suppressed vasodilator responses to transmural field stimulation. The effect of bradykinin was unaltered by indomethacin. N omega-Nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthesis, abolished the inhibition of vasodilator responses to transmural field stimulation elicited by bradykinin. However, in the presence of L-arginine and L-NAME the inhibitory effect of bradykinin reappeared. Furthermore, methylene blue itself caused potentiation of vasodilator responses to transmural field stimulation and reversed the effect of bradykinin. These findings suggest that bradykinin can produce an inhibitory modulation of the actions of sensory nerves in the perfused rat mesentery and that the effect of bradykinin may be mediated by nitric oxide released from non-adrenergic, non cholinergic nerves. PMID- 8223923 TI - Effects of cyclopiazonic acid on Ca(2+)-activated tension production in skinned skeletal muscle fibres of the ferret. AB - The effect of cyclopiazonic acid, an inhibitor of sarcoplasmic reticulum Ca(2+) ATPase, was investigated on force generation by the contractile apparatus in Triton-skinned fibres from extensor digitorum longus and soleus. Concentrations of cyclopiazonic acid lower than 10 microM were without effect on Ca(2+) activated tension in both types of muscles. In contrast, in soleus, cyclopiazonic acid (20, 50, 100 microM) was found to shift reversibly the relation-tension pCa (-log[Ca2+]) towards lower free Ca2+ and to decrease maximal Ca(2+)-activated tension, in a dose-dependent manner. In extensor digitorum longus, the Ca2+ sensitivity was significantly increased only at a high cyclopiazonic acid concentration (100 microM) and for all the concentrations tested between 5 to 100 microM maximal Ca(2+)-activated tension was unchanged. These results suggest that cyclopiazonic acid has a direct effect on contractile proteins, in a dose dependent manner. Ca2+ sensitivity and Ca(2+)-activated maximal tension of the contractile apparatus were differentially affected in fast- (extensor digitorum longus) and slow-twitch (soleus) fibres from ferret skeletal muscle. PMID- 8223924 TI - The effects of different antiemetic agents on morphine-induced emesis in ferrets. AB - There is interest in the development of antiemetics other than dopamine receptor antagonists for the treatment of postoperative nausea and vomiting. A ferret model of morphine-induced emesis may have wider application in evaluating newer agents than the apomorphine dog model. This study describes the conditions for morphine-induced emesis in ferrets and evaluates five antiemetics that are prototypical of three different mechanisms. The average numbers of vomiting and retching episodes induced by morphine (0.1-2.5 mg/kg s.c.) were distributed as a bell-shaped curve. Maximum number of vomits occurred at 0.3 mg/kg (11.8 +/- 2.1 vomits; 45 +/- 12.5 retches). Antiemetics or vehicle were given i.v. 5 min prior to morphine while each ferret was maintained under isoflurane-O2 anesthesia. Ondansetron, a 5-HT3 receptor antagonist, reduced vomiting episodes by 47% and 70% (3 and 10 mg/kg). Granisetron, a 5-HT3 receptor antagonist was inactive at doses of 0.1, 1.0, 3.0 and 10 mg/kg. Metoclopramide reduced vomiting episodes by 48% and 82% (3 and 10 mg/kg). Droperidol reduced vomiting episodes by 84% at 3 mg/kg. Naloxone reduced vomiting episodes by 91% and 43% at doses of 0.1 and 1.0 mg/kg. In most cases, prolonged latency times to the first episodes accompanied the reduction in total numbers of episodes. The significant reduction of morphine induced emesis in the ferret by ondansetron, metoclopramide and droperidol is consistent with the reduction of postoperative emesis in man by these compounds when morphine was a component of the anesthetic regimen. These results suggested that the morphine ferret model may be useful for evaluating compounds having the potential for preventing and treating postoperative vomiting. PMID- 8223925 TI - Evidence that ATP, ADP and AMP are not ligands of the striatal adenosine A2A receptors. AB - It has been claimed recently that, in several cell types, ATP can induce a stimulation of cAMP production which is sensitive to methylxanthine inhibition and is not mediated by the ATP degradation product, adenosine. One explanation for these results would be direct activation of adenosine A2 receptors by ATP itself. We have therefore investigated whether adenine nucleotides are ligands of adenosine A2A receptors from bovine striatum. We show here that ATP, ADP, AMP and their phosphorothioates analogues (ATP gamma S, ADP beta S and AMP alpha S), at a 100 microM concentration, produced a 83-91% inhibition of the binding of [3H]CGS21680, an adenosine A2A receptor agonist, to striatum membranes. However, this action was inhibited by adenosine deaminase or by adenosine 5'-O-(alpha, beta-methylene)diphosphate (APCP), an inhibitor of 5'-nucleotidase-mediated AMP degradation. The effects of adenosine deaminase and APCP were dependent on their concentration. These results indicate that ATP, ADP and even AMP can exert an effect on the adenosine A2A receptors only through their breakdown into adenosine by ectonucleotidases. PMID- 8223926 TI - Partial antinociceptive cross-tolerance to intracerebroventricular beta-endorphin in mice tolerant to systemic morphine. AB - The effects of subcutaneous morphine pellet-implantation on antinociception induced by intracerebroventricular (i.c.v.) administration of beta-endorphin or morphine and intrathecal (i.t.) administration of morphine, [D-Pen2,D Pen5]enkephalin (DPDPE), [D-Ala2,NMePhe4,Gly5-ol]enkephalin (DAMGO), serotonin or norepinephrine were studied in male ICR mice. The tail-flick and hot-plate responses were used for antinociceptive tests. The ED50 values for i.c.v. administered morphine for antinociception in morphine pellet-implanted mice were increased from 3.3- and 2.2-fold at 0 h to 14.2- and 19.0-fold at 4 h and declined to 4.8- and 3.0-fold at 8 h after pellet removal in the tail-flick and hot-plate tests, respectively. On the other hand, the ED50 values for i.c.v. administered beta-endorphin for antinociception were only slightly increased (1.7 to 5.1-fold increases) throughout the same time course. The inhibition of the tail-flick response induced by i.t. injection of morphine, DPDPE and serotonin, but not norepinephrine or DAMGO, was attenuated in morphine pellet-implanted mice. These findings are consistent with previous studies indicating that different neuronal mechanisms are involved in morphine- and beta-endorphin induced antinociception. PMID- 8223927 TI - Delayed regional metabolic actions of phencyclidine. AB - Phencyclidine (PCP), a psychotomimetic drug of abuse, produces mental changes and manifestations in humans which are reminiscent of schizophrenia, though the mechanism of these actions remains unknown. We report here a biphasic time course of PCP action on regional cerebral glucose metabolism extending over 48 h. A single dose of PCP (8.6 mg/kg) produces an initial increase in glucose metabolism (at 3 h) and a later decrease in glucose metabolism (at 24 h) without a return to baseline until 48 h. A single lower dose of PCP (0.86 mg/kg), a dose which is considered selective for action at the NMDA-PCP receptor, produces no early metabolic change (at 3 h), but replicates the regional hypometabolism albeit less intense at 24 h. The delayed cerebral hypometabolism does not appear to be related to PCP-induced intracellular vacuolization, seen in the retrosplenial cortex. These metabolic changes may be associated with the psychotomimetic effects of PCP and thus may be relevant to psychosis in humans. PMID- 8223928 TI - Protective effect of SV-IV on platelet-activating factor-induced hypotension, bronchoconstriction and gastric mucosal injury. AB - The inhibitory effect of one of the major proteins secreted by the rat seminal vesicles (SV-IV) on platelet-activating factor (PAF)-induced biological activities was investigated in vivo. SV-IV was found to prevent dose dependently both hypotension and acute bronchospasm caused by PAF administration in guinea pigs. In addition, SV-IV inhibited both PAF- and ethanol-induced gastric mucosal injury in a dose-dependent manner. PMID- 8223929 TI - Endothelium-independent pressor effect of big endothelin-1 and its inhibition by phosphoramidon in rat mesenteric artery. AB - We asked whether or not the endothelium plays a functional role in the conversion of big endothelin-1 to endothelin-1 in the perfused rat mesenteric artery. In endothelium-denuded preparations, big endothelin-1 produced a much more potent pressor effect than in intact preparations. Phosphoramidon suppressed the big endothelin-1-induced pressor action without affecting the action of endothelin-1, irrespective of the presence or absence of the endothelium. The amounts of immunoreactive-endothelin in the perfusate during perfusion of endothelium denuded preparations with big endothelin-1 were extremely low compared with those observed in intact preparations and were not significantly suppressed by the metalloproteinase inhibitor, phosphoramidon, in contrast to the case with intact preparations. When synthetic endothelin-1 was perfused in the endothelium-denuded mesentery, the peptide disappeared from the perfusate more rapidly than with intact preparations, suggesting that endothelin-1 generated from big endothelin-1 is effectively trapped by vascular smooth muscle cells in the endothelium-denuded preparation. Our results suggest that the endothelium is not essential for the conversion of big endothelin-1 to endothelin-1, in rat mesenteric artery. PMID- 8223930 TI - Indication of methamphetamine-induced reactive synaptogenesis in the prefrontal cortex of gerbils (Meriones unguiculatus). AB - A single dose of methamphetamine (25 mg/kg i.p.) was administered to young adult gerbils (Meriones unguiculatus) aged 90 days and the number of spices was determined along 40-microns segments of basal, lateral and apical dendrites of pyramidal cells in layers III and V of the prefrontal cortex, after 1.5, 7, 20 and 30 days. The density of spines rapidly increased by more than 80% within 7 days after drug challenge, and subsequently returned to the original normal values within about 2 weeks. Thirty days after drug administration the density of dendritic spines was slightly, but significantly, less than control values (about 5%). The density of spines was likewise affected in layer III and V neurones, irrespective of the spatial domain of their dendritic ramifications. Since several lines of investigation indicate that methamphetamine can cause the destruction of dopaminergic nerve terminals in the mammalian forebrain, the present results are discussed against the background of current concepts about reactive synaptic reorganization and adaptive remodelling of neural circuits in the central nervous system. PMID- 8223931 TI - Antagonism by citalopram and tianeptine of presynaptic 5-HT1B heteroreceptors inhibiting acetylcholine release. AB - The interactions of citalopram and tianeptine, two antidepressants having opposite effects on serotonin (5-HT) uptake, with 5-HT1B presynaptic heteroreceptors located on cholinergic terminals were investigated. In rat hippocampal synaptosomes, citalopram (0.01 or 0.1 microM) or tianeptine (0.01-10 microM) did not modify the basal or the K(+)-evoked release of [3H]acetylcholine. Only at the concentration of 100 microM did tianeptine significantly decrease ( 18%) the K(+)-evoked release of [3H]acetylcholine without affecting the spontaneous outflow of radioactivity. The inhibitory effect of 7-trifluoromethyl 4-(4-1-piperazinyl)-pyrrolo[1,2-a]quinoxaline (CGS 12066B), a 5-HT1B receptor agonist, on the stimulation-induced release of [3H]acetylcholine was reduced in a concentration-dependent manner by citalopram and tianeptine. Both drugs completely reversed the inhibitory effects of CGS 12066B at concentrations that did not modify by themselves the release of [3H]acetylcholine. In contrast, tianeptine, up to a concentration of 1 microM, failed to antagonise the inhibitory effect of the muscarinic receptor agonist carbachol on K(+)-evoked [3H]acetylcholine release. Finally, the administration of tianeptine ex vivo (10 or 20 mg/kg) modified neither the depolarisation-induced release of [3H]acetylcholine nor the inhibitory effect of CGS 12066B on this presynaptic process. These findings further confirm that antidepressants interact in vitro with presynaptic 5-HT1B heteroreceptors. PMID- 8223932 TI - In vivo reactivity of resistance arterioles to activation of ATP-sensitive K+ channels. AB - Our goal was to examine in vivo responses of resistance arterioles contained within the hamster cheek pouch microcirculation to activation of ATP-sensitive potassium (K+) channels. We measured changes in diameter of cheek pouch arterioles in response to activation of ATP-sensitive K+ channels using RP 52891 (Aprikalim) and BRL 38227 (active enantiomer of cromakalim). RP 52891 and BRL 38227 produced dose-related dilatation of arterioles, which was inhibited by glibenclamide (1.0 microM), but not altered by NG-monomethyl-L-arginine (L-NMMA; 1.0 microM). Glibenclamide did not alter baseline diameter of cheek pouch arterioles and did not alter dilatation of cheek pouch arterioles in response to nitroglycerin (1.0 and 10 microM). L-NMMA did not alter dilatation of arterioles to nitroglycerin, but inhibited dilatation of arterioles to acetylcholine (0.1, 1.0 and 10 microM). Thus, dilatation of arterioles in response to activation of ATP-sensitive K+ channels appears to be specific and does not involve the release of nitric oxide or a nitric oxide containing compound. The findings of the present study suggest that ATP-sensitive K+ channels are functional in resistance arterioles in vivo, but do not appear to affect resting tone of cheek pouch arterioles. PMID- 8223933 TI - Simultaneous measurement of vasodilation and changes in cyclic nucleotides in the perfused mesenteric arterial bed of the rat. AB - We examined the relationship between relaxation responses of the mesenteric arterial bed and the levels of cAMP and cGMP released from the rat mesenteric arterial bed. Perfusions of mesentery preparations with 1 microM acetylcholine, 0.1 microM calcium ionophore A23187, and 1 microM sodium nitroprusside all produced complete and long-lasting relaxation and increased the levels of cAMP as well as cGMP in the effluent. In endothelium-denuded preparations, acetylcholine did not elicit either vasorelaxation or an increase in cAMP and cGMP levels. Perfusion of the endothelium-denuded preparation with 1 microM sodium nitroprusside evoked complete relaxation and a marked increase in cGMP levels but not cAMP levels. Isoproterenol (1 microM) produced complete relaxation and an increase in cAMP levels, but did not affect cGMP levels either in the preparation with or in that without endothelium. Acetylcholine (0.001-1 microM) relaxed the preparation and increased cAMP and cGMP levels in the effluent in a dose dependent manner. The acetylcholine-induced relaxation was reversed by 45% following perfusion with 10 microM methylene blue, and both the cAMP and cGMP levels were decreased. L-NG-Monomethyl arginine (L-NMMA) (100 microM), a nitric oxide synthase inhibitor, completely reversed the relaxation induced by 0.1 microM acetylcholine and reduced the elevated cGMP levels. Indomethacin (1 microM) reduced the acetylcholine-induced cAMP release, but did not alter the vasorelaxation in response to acetylcholine. We propose a novel method for the simultaneous measurement of vasodilation and changes in cAMP and cGMP levels released from the rat mesenteric arterial bed. We conclude that this method may provide information about the function of the endothelium of resistance vessels. PMID- 8223934 TI - Characterization of BIBN 99: a lipophilic and selective muscarinic M2 receptor antagonist. AB - The present study was designed to characterize the receptor selectivity profile of the novel muscarinic M2 receptor antagonist BIBN 99 (5,11-dihydro-8-chloro-11 [[4-[3-[(2,2-dimethyl-1- oxopentyl)ethylamino]propyl]-1-piperidinyl]acetyl]-6H- pyrido[2,3-b][1,4]benzodiazepin-6-one). In radioligand binding studies BIBN 99 showed high affinity for m2/M2 sites (pKi = 7.52/7.57), intermediate affinity for m4 sites (pKi = 6.76) and low affinity for m1/M1 (pKi = 5.97/6.17), m3/M3 (pKi = 6.11/6.04) and m5 sites (pKi = 5.84). Functional studies in vitro showed BIBN 99 to be a competitive antagonist and to have an 11- to 25-fold higher affinity for M2 receptors than for putative M1 receptors in the rabbit vas deferens or M3 receptors in guinea-pig trachea. In vivo studies revealed that BIBN 99 is able to cross the blood-brain barrier, and although showing an approximately 3-fold higher affinity for M2 binding sites BIBN 99 appeared to be 7- to 18-fold less potent than AF-DX 116 in inhibiting muscarinic agonist or vagally induced bradycardia in rats and guinea-pigs. The results show that BIBN 99 is the first lipophilic muscarinic M2 receptor antagonist to have remarkable M2 versus M1 selectivity (30-fold). In addition, BIBN 99 possesses central nervous system activity and only minor peripheral cardiac effects. PMID- 8223935 TI - Role of the nucleus tractus solitarii and the rostral depressive area in the sympatholytic effect of 8-hydroxy-2-(di-n-propylamino)tetralin in the cat. AB - Bilateral microinjections of kainic acid (500 ng/site) into the nucleus tractus solitarii produced hypertension, tachycardia and sympatho-excitation in anesthetized cats. The cardiac-related component of renal sympathetic nerve activity was abolished as well as the sympatho-inhibitory effects that accompany the phenylephrine (5-10 micrograms/kg i.v.)-induced hypertension. About 60 min after kainic acid microinjections into the nucleus tractus solitarii, 8-hydroxy-2 (di-n-propylamino)tetralin (8-OH-DPAT) administered in cumulative doses (1-100 micrograms/kg i.v.) failed to alter mean blood pressure, heart rate or renal sympathetic nerve activity. In addition, bilateral microinjections of 8-OH-DPAT (2 nmol in 40 nl) into the nucleus tractus solitarii did not change mean blood pressure, heart rate or renal sympathetic nerve activity. Microinjections of kainic acid into the rostral vasodepressive area produced hypotension, bradycardia and renal sympatho-inhibition followed by persistent increases in blood pressure, heart rate and renal sympathetic nerve activity. These effects were also associated with an inhibition of the baroreceptor reflex elicited by phenylephrine and by the disappearance of the synchronism between the renal sympathetic bursts and cardiac rhythm. Subsequent i.v. 8-OH-DPAT (1-100 micrograms/kg) elicited decreases in mean blood pressure, heart rate and in renal sympathetic nerve activity. Central baroreceptor denervation by kainic acid lesions of the lateral tegmental field largely attenuated the hypotensive, bradycardiac and sympatho-inhibitory effects elicited by 8-OH-DPAT applied to the ventral surface of the rostral ventrolateral medulla.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223936 TI - Antinociceptive activity of intrathecal ketorolac is blocked by the kappa-opioid receptor antagonist, nor-binaltorphimine. AB - Systemic and intrathecally administered ketorolac produced antinociception in the p-phenylquinone test, but not in the tail-flick or hot-plate tests. Antagonists of the subtypes of opioid receptors were used to evaluate the interaction of ketorolac with these receptors. Intrathecally administered kappa-opioid receptor antagonist nor-binaltorphimine dihydrochloride blocked the antinociceptive effects of systemic ketorolac and intrathecally administered ketorolac. Naloxone and ICI 174,864 failed to block the effects of ketorolac. Activation of nor binaltorphimine-sensitive receptors appears to be an integral element in the mechanism of antinociception of ketorolac at the spinal level. Ketorolac did not precipitate withdrawal jumping in morphine-tolerant mice demonstrating that ketorolac does not act as a mixed agonist-antagonist at the opioid receptor. We suggest that neuraxial placement of ketorolac may prove useful in the clinical setting for the management of acute pain in humans. PMID- 8223937 TI - Tolbutamide, but not glyburide, affects the excitability and contractility of unfatigued frog sartorius muscle. AB - The goal of this study was to characterize the effects of tolbutamide and glyburide, two known KATP channel blockers, on intact, unfatigued sartorius muscle fibres of the frog, Rana pipiens. Tetanic contractions were elicited by field stimulation with 200 ms long train of pulses (0.5 ms, 6 V, 140 Hz). Resting and action potentials were measured using conventional microelectrodes. At pHo 7.2 (extracellular pH), the tetanic force was unaffected by 0.5 mM and 1.0 mM tolbutamide, but at 2.0 mM it decreased by 15.5 +/- 1.0%. The effect of tolbutamide on the tetanic force was significantly greater at pHo 6.4: all three tolbutamide concentrations caused a significant decrease in tetanic force, being 62.3 +/- 9.4% at 2 mM. In the presence of tolbutamide a large number of fibres became unexcitable at pHo 6.4, but not at pHo 7.2. Glyburide at 10 microM, on the other hand, caused a 5-7% decrease in tetanic force at both pHo 6.4 and 7.2, but no further decreases in tetanic force were observed when the glyburide concentration was increased up to 100 microM. Unlike tolbutamide, glyburide did not affect the excitability of muscle fibres, but significantly prolonged the repolarization phase of action potentials, especially at pHo 6.4. We suggest that several of the tolbutamide effects reported in this study cannot be accounted for by a direct effect on KATP channels, and that the large decrease in membrane excitability and muscle contractility in the presence of tolbutamide must seriously be taken into consideration when this channel blocker is used to study the physiological role of KATP channels in intact muscle fibres. PMID- 8223938 TI - Effect of morphine on urine output: possible role of atrial natriuretic factor. AB - We examined the possible role of atrial natriuretic factor (ANF) in morphine induced diuresis. Morphine (20 micrograms/20 microliters) administered intracerebroventricularly (i.c.v.) in conscious, normally hydrated Sprague-Dawley rats significantly increased urine output from 0.22 +/- 0.16 ml/h/rat to 2.07 +/- 0.46 ml/h/rat, 3 h after injection. Morphine injection also significantly increased the plasma ANF level to a maximum of 364 +/- 46 pg/ml compared to control animals (33 +/- 7 pg/ml) in which 20 microliters of saline was injected i.c.v. (P < 0.001). This peak increase occurred 1 h after morphine administration, however, it continued to remain significantly elevated (171 +/- 36 pg/ml vs. 39 +/- 8 pg/ml in control animals; P < 0.001) 2 h later. The maximal diuresis observed 3 h after administration of morphine i.c.v. was abolished by pretreatment of the animals with 0.8 mg of naloxone given intravenously (i.v.) and also by anti-rat ANF serum (0.4 ml i.v.). A short, transient increase in blood pressure, occurred 2-5 min after morphine administration, but it is unlikely that this increase accounted for the increase in plasma ANF. We conclude that the diuresis induced by i.c.v. administration of morphine is due to enhanced release of ANF. PMID- 8223939 TI - 5-HT1 receptors mediating contraction in bovine cerebral arteries: a model for human cerebrovascular '5-HT1D beta' receptors. AB - We report on the pharmacological profile of the 5-HT receptor which induces contraction of the bovine isolated cerebral arteries. Several 5-HT receptor agonists were tested for their ability to induce vasoconstriction in bovine pial arteries and their potencies were compared to that of 5-HT. The rank order of agonist potency can be summarized as 5-carboxamidotryptamine (5-CT) = RU 24969 > or = 5-HT > sumatriptan > alpha-methyl-5-HT > methysergide > 2-methyl-5-HT > ((+ )-2-dipropylamino-8-hydroxy-1,2,3,4-tetrahydronaphthalene (8-OH-DPAT). Only methysergide induced a contraction which was smaller than that elicited by 5-HT. Antagonists with selective affinity at 5-HT1A/1B (propranolol), 5-HT1C (mesulergine), 5-HT2 (ketanserin, mianserin) and 5-HT3 (MDL 72222) sites were inactive to block the 5-HT-induced contraction. In contrast, the 5-HT1/5-HT2 receptor antagonists methiothepin and metergoline inhibited the 5-HT-induced response with relatively high affinity (pA2 = 8.16 +/- 0.26 and 6.73 +/- 0.05, respectively). Overall, this pharmacological profile indicated clearly that a 5 HT1 receptor, most closely related to the 5-HT1D subtype, is responsible for the 5-HT-induced contraction of bovine cerebral arteries. Correlation analysis of the potencies of a series of 5-HT receptor agonists and antagonists in bovine and human cerebrovascular preparations showed a highly significant positive correlation (r = 0.94, P = 0.0051).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223940 TI - d-Fenfluramine- and d-norfenfluramine-induced hypophagia: differential mechanisms and involvement of postsynaptic 5-HT receptors. AB - Severe depletion of 5-hydroxytryptamine (5-HT) by para-chlorophenylalanine (pCPA, 150 mg/kg per day x3) did not alter the hypophagic effect of d-fenfluramine (1-3 mg/kg i.p.) 1 h after food presentation in 24-h food-deprived rats, and moderately and comparably increased the hypophagic effects of its metabolite, d norfenfluramine (0.35-1.0 mg/kg i.p.), and of the 5-HT1C receptor agonist, 1-(3 chlorophenyl)piperazine (mCPP; 1.5, 2.0 mg/kg i.p.). Chronic treatment with mCPP (2.5 mg/kg i.p. x 14) attenuated the hypophagia induced by d-norfenfluramine (1, 1.5 mg/kg) but not d-fenfluramine (1, 3 mg/kg). 1-(1-Naphthyl)piperazine (3, 8 mumol/kg s.c.), which has greater affinity for 5-HT1C than for 5-HT2 receptors, had no effect on the hypophagia induced by d-fenfluramine (1.25, 2.0 mg/kg), but 1.3 and 3 mumol/kg 1-(1-naphthyl)piperazine largely and comparably attenuated the substantial hypophagic effect of d-norfenfluramine (0.75 mg/kg). The essentially complete hypophagic action of d-norfenfluramine (1.25 mg/kg) was inhibited by 1 (1-naphthyl)piperazine with ID50 = 2.13 mumol/kg. Ketanserin, which binds more weakly than 1-(1-naphthyl)piperazine to 5-HT1C receptors and more strongly to 5 HT2 receptors, attenuated weaker but not stronger hypophagic effects of d fenfluramine (1.25, 2.0 mg/kg) when given at high dosage (8, 16 mumol/kg s.c.). Ketanserin (16 mumol/kg) also weakly attenuated the hypophagia due to d norfenfluramine (0.75 mg/kg), but not the essentially complete hypophagia due to d-norfenfluramine (1.25 mg/kg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223941 TI - Renal effects of neutral endopeptidase inhibition in euvolemic and hypervolemic rats. AB - Candoxatrilat is a potent and selective inhibitor of neutral endopeptidase (EC 3.4.24.11), the enzyme responsible for the degradation of atrial natriuretic factor (ANF). In these studies, the renal effects of candoxatrilat were investigated in euvolemic and hypervolemic anaesthetised rats. In euvolemic rats, candoxatrilat (675 micrograms/kg per h) had no effect on urine output, sodium and potassium excretion or urinary cyclic GMP excretion. However, in hypervolemic rats, the natriuretic and diuretic responses to volume expansion were markedly potentiated by the candoxatrilat infusion, with a concomitant increase in urinary cyclic GMP. Acute volume expansion was characterised by natriuresis, diuresis and increased levels of plasma ANF and cyclic GMP (1.5-fold and 2-fold increases respectively, when compared to euvolemic rats). The results presented suggest that plasma ANF levels and volume status modulate responses to neutral endopeptidase inhibition. The development of the neutral endopeptidase inhibitor, candoxatrilat, provides the opportunity to exploit endogenous ANF effectively in disease states with elevated ANF. PMID- 8223942 TI - Effects of the novel 5-HT1A receptor antagonist, (+)-WAY 100135, on stereotyped behaviour induced by the NMDA receptor antagonist dizocilpine in rats. AB - We have recently shown that the stereotyped behaviour induced by the uncompetitive NMDA receptor antagonist dizocilpine (MK-801) can be attenuated or blocked by partial agonists at 5-HT receptors of the 5HT1A subtype, indicating that 5-HT (5-hydroxytryptamine, serotonin) is involved in the stereotyped behaviour produced by dizocilpine. In the present experiment, a selective, silent 5-HT1A receptor antagonist, (+)-WAY 100135 (N-tert-butyl 3-4(2-methoxyphenyl) piperazin 1-yl-2-phenylpropanamide dihydrochloride), was used to further study the role of 5-HT activation in dizocilpine-induced behaviours. At a dose of 10 mg/kg, (+)-WAY 100135 significantly reduced the intensity of head weaving induced by dizocilpine, but this effect was lost by increasing the dose to 20 mg/kg. At this higher dose, (+)-WAY 100135 induced marked but short-lasting increases in skeletal muscle tone and hindlimb abduction, resembling components of the '5-HT behavioural syndrome', which would explain its biphasic effects on dizocilpine induced behaviours. The data substantiate that, in addition to the well known activation of dopaminergic transmission, activation of the 5-HT system might be involved in the behavioural effects of NMDA receptor antagonists such as dizocilpine. PMID- 8223943 TI - In vitro characterization of a novel Ca2+ entry blocker: SR 33805. AB - In this study, SR 33805 was shown to inhibit competitively [3H]fantofarone binding to cardiac sarcolemmal membranes. In contrast, SR 33805 was shown to inhibit allosterically [3H](+)-PN200-110, [3H](-)-D888 and cis-(+)-[3H]diltiazem binding. In isolated rabbit atrial preparations, SR 33805 was shown to be the least potent of fantofarone, nifedipine, verapamil and diltiazem in terms of both negative chronotropic and inotropic responses (IC50's 6 and 12 microM, respectively). In superfused rat aortic strips, SR 33805 like other Ca2+ channel antagonists, caused a significant inhibition of both K(+)-induced 45Ca2+ influx and contractile responses. In addition this agent was shown to antagonize Ca(2+) induced contractions in K(+)-depolarized aorta with a pA2, value of 8.39 +/- 0.02. In femoral, renal and basilar arteries, SR 33805 was equiactive to the other Ca2+ channel antagonists studied in antagonizing K(+)-induced contractions (IC50 approximately 40 nM), but unlike the reference Ca2+ channel antagonists, was equiactive in antagonizing serotonin-induced contractions (IC50 approximately 250 nM). This suggests that the effects of SR 33805 depend mainly on membrane potential. In conclusion, SR 33805 is a potent Ca2+ channel antagonist which, unlike fantofarone, verapamil and diltiazem, is highly selective for vascular smooth muscle and devoid of any potent negative inotropic actions. PMID- 8223944 TI - A novel non-NMDA receptor antagonist shows selective displacement of low-affinity [3H]kainate binding. AB - 5-Nitro-6,7,8,9-tetrahydrobenzo[G]indole-2,3-dione-3-oxime (NS-102), a new competitive glutamate receptor antagonist displaced binding to non-N-methyl-D aspartate (non-NMDA) binding sites with no activity at the NMDA and strychnine insensitive glycine binding sites. Under experimental conditions in which both high- and low-affinity sites were labelled, NS-102 only partially inhibited the binding of [3H]kainate. Studies of NS-102 displacement of high-affinity versus low-affinity [3H]kainate binding showed a high selectivity of NS-102 for the low affinity [3H]kainate binding site (Ki = 0.6 microM) compared to the high-affinity [3H]kainate binding site (Ki > 10 microM). NS-102 was a relatively weak inhibitor of 2-amino-3-(3-hydroxy-5-methylisoxazol-4-yl)propionic acid (AMPA) binding (IC50 = 7.2 microM). NS-102 and related compounds with similar pharmacological profiles may become valuable tools in the characterization of the functional importance of the low-affinity [3H]kainate binding site. PMID- 8223945 TI - Two different mRNAs from rat liver code for the transport of bumetanide and taurocholate in Xenopus laevis oocytes. AB - The aim of this study was to elucidate whether bumetanide, which is a competitive inhibitor of carrier mediated bile acid uptake in liver cells, is transported by bile acid carriers. The expression of hepatocellular transport proteins for bile acid uptake and the uptake of the loop diuretic bumetanide was therefore studied in Xenopus laevis oocytes by injection of rat liver poly(A)(+)-RNA. Three hours after injection, a 70% increase in [3H]taurocholate uptake versus noninjected oocytes was accompanied by an increase in only 24% in the uptake of [3H]bumetanide. Size fractionation of the poly(A)(+)-RNA yielded 33 mRNA fractions of which fraction 21 accounted for an 800% increase of taurocholate transport with only a slight increase in bumetanide uptake. Bumetanide transport was coded by mRNA-fraction 18, which stimulated uptake by 160-200% with a concomitant small increase in taurocholate uptake. Uptake of cholate was induced by both mRNA fractions with almost 2.5 fold greater expression by the bumetanide fraction. Oocyte transport of taurocholate (expressed by fraction 21) and bumetanide transport (expressed by fraction 18) were characterized in terms of Na+ dependency, inhibition by 4,4'-diisothiocyano-1,2-diphenylethane-2,2' disulfonic acid (DIDS) and mutual competition. The results indicate that the bumetanide transporter mRNA is clearly different from the mRNA for the taurocholate transport protein. The mRNA fraction 18 was used for the construction of a cDNA library. PMID- 8223946 TI - Chronic opioid treatment may uncouple opioid receptors and G-proteins: evidence from radiation inactivation analysis. AB - Radiation inactivation (target size analysis) was used in this study to determine whether uncoupling of opioid receptor and G-protein is a contributing mechanism to opioid tolerance. Male Sprague-Dawley rats (160-260 g) were rendered tolerant to morphine or [D-Ala2,D-Leu5]enkephalin (DADLE) by multiple i.p. or i.c.v. injections twice a day for 6 or 5 days. Control rats were injected with saline instead of opioids. The animals were killed, the midbrains excised and pooled together for each group. The washed P2 membranes were suspended in buffer and irradiated with 1-10 Mrad doses of 60Co irradiation, following which mu- or delta opioid receptor binding activity of each sample was assayed. The molecular weight of the receptor was calculated from a standard irradiation curve constructed using several enzyme markers of known molecular weight. We found that the functional molecular size of mu-opioid receptor significantly decreased from 349 kDa to 228 kDa after 6 days of chronic morphine treatment, while, the molecular size of delta-opioid receptor decreased from 303 kDa to 223 kDa after 5 days of chronic DADLE treatment. These results are consistent with the uncoupling of opioid receptor from G-protein during chronic opioid treatment. PMID- 8223947 TI - Dual activation of GABAA and glycine receptors by beta-alanine: inverse modulation by progesterone and 5 alpha-pregnan-3 alpha-ol-20-one. AB - The differential sensitivity of the glycine and GABAA receptors to modulation by progesterone and 5 alpha-pregnan-3 alpha-ol-20-one (5 alpha 3 alpha) was used to determine whether beta-alanine acts through its own receptor, or through the glycine and/or GABAA receptor(s). The response to beta-alanine resembles the glycine response as it is inhibited by strychnine (a competitive glycine antagonist) or progesterone (a negative modulator of the glycine response). Significantly, the response to beta-alanine also resembles the GABA response in that it is inhibited by 2-(carboxy-3'-propyl)-3-amino-6-paramethoxy phenylpyridazinium+ ++ bromide (SR-95531; a competitive GABA antagonist) and potentiated by 5 alpha 3 alpha (a positive modulator of the GABA response). The efficacy of beta-alanine at the GABAA receptor is comparable to that of GABA. Similarly, the efficacy of beta-alanine at the glycine receptor is comparable to that of glycine. The greater potency of beta-alanine at the glycine receptor indicates that, if beta-alanine is a neurotransmitter, its effects are more likely to be mediated by glycine receptors than by GABAA receptors. However, activation of the GABAA receptor by beta-alanine may become important in the presence of steroid modulators such as progesterone or 5 alpha 3 alpha. PMID- 8223948 TI - Partial and full agonists/inverse agonists affect [35S]TBPS binding at different occupancies of central benzodiazepine receptors. AB - Concentration-dependent effects of benzodiazepine receptor ligands were examined on nonequilibrium binding of t-butylbicyclophosphoro[35S]thionate (TBPS, 20 min of incubation at 25 degrees C) to synaptosomal membranes of rat cerebral cortex. Benzodiazepine receptor occupancies were calculated from the displacing potencies of the ligands determined for [3H]flumazenil binding under identical conditions. Greater maximal enhancing (i.e. accelerating) effects of the full agonists diazepam and flunitrazepam on [35S]TBPS binding were reached at lower occupancies of benzodiazepine receptors than the smaller enhancing effects of the partial agonists bretazenil and the beta-carboline ZK 91296. Similarly, the maximal decreasing effect of the full inverse agonist methyl 6,7-dimethoxy-4-ethyl-beta carboline-3-carboxylate (DMCM) on TBPS binding was reached at lower occupancy than that of the partial inverse agonist FG 7142. Half-maximal effects on TBPS binding corresponded to about 20-30% occupancies for the full agonists and DMCM, while for partial agonists and FG 7142 they exceeded 60-80% occupancies. Different (concave versus convex) shapes of the occupancy-effect curves can also differentiate partial from full agonists and inverse agonists. The results suggest that different pharmacological efficacies of benzodiazepine receptor ligands are associated with differences in coupling between benzodiazepine and convulsant binding sites to modulate the chloride ionophores. PMID- 8223949 TI - Effects of HOE 694--a novel inhibitor of Na+/H+ exchange--on NIH 3T3 fibroblasts expressing the RAS oncogene. AB - Among the sequelae of ras oncogene expression are intracellular alkalinization and increase of cell volume, both phenomena attributed at least in part to activation of the Na+/H+ exchanger. The present study was performed to elucidate the effects of HOE 694--a novel inhibitor of the Na+/H+ exchanger--on intracellular pH, cell volume, cytoarchitecture and cell proliferation of ras oncogene expressing NIH 3T3 fibroblasts. Following transient exposure of the cells to 20 mmol/l NH3/NH4+, intracellular pH decreases sharply. The following slow realkalinization is completely blocked by 10 mumol/1 HOE 694. Half-maximal inhibition is achieved by 100 nmol/l HOE 694. Cell proliferation is inhibited by HOE 694 with similar potency, whereas the increase in cell volume and cytoskeletal transformation are not prevented, even by 10 mumol/l HOE 694. PMID- 8223950 TI - Rat vena cava alpha 1B-adrenoceptors: characterization by [3H]prazosin binding and contraction experiments. AB - We examined which subtypes of alpha 1-adrenoceptors are expressed in rat vena cava by using both functional and [3H]prazosin binding experiments. Pretreatment with chloroethylclonidine inactivated about 80% of the specific [3H]prazosin binding sites and reduced the maximal noradrenaline-induced contraction to the same extent. Competition with subtype-selective agonists and antagonists showed primarily the alpha 1B-adrenoceptor subtype in vena cava. The number of alpha 1 adrenoceptors estimated with [3H]prazosin binding and the maximal noradrenaline induced contraction were dose-dependently inhibited by phenoxybenzamine, indicating the absence of receptor reserve for noradrenaline in vena cava. As the noradrenaline-induced contraction was largely inhibited in Ca(2+)-free solution, these results suggest that alpha 1B-adrenoceptors can be mainly linked to Ca2+ influx in rat vena cava. PMID- 8223951 TI - Expression and properties of recombinant alpha 1 beta 2 gamma 2 and alpha 5 beta 2 gamma 2 forms of the rat GABAA receptor. AB - The interaction of omega (benzodiazepine) modulatory drugs with transiently expressed alpha 1 beta 2 gamma 2 and alpha 5 beta 2 gamma 2 forms of the rat GABAA receptor was investigated using [3H]flumazenil as a probe in in vitro radioligand binding assays. The imidazopyridines alpidem and zolpidem exhibited pronounced selectivity for the alpha 1- compared to the alpha 5-containing construct, whereas omega (benzodiazepine) site modulatory compounds from other chemical series including diazepam, tetrazepam, zopiclone, triazolam, bretazenil and midazolam behaved as relatively non-selective drugs. In the presence of 10 microM gamma-aminobutyric acid (GABA) the potencies of diazepam, flunitrazepam and midazolam to inhibit [3H]flumazenil binding to the alpha 1-construct were increased 3 to 5 fold, whereas with 6,7-dimethoxy-4-ethyl-beta-carboline-3 carboxylate methyl ester a 2.5-fold reduction in potency was observed. Similar modulatory effects of GABA were obtained with these drugs, using the alpha 5 construct. We suggest that these GABA shift determinations of [3H]flumazenil binding can be used as a rapid test to evaluate the intrinsic activities of omega modulatory compounds. PMID- 8223952 TI - Binding of BW 373U86, a non-peptidic delta opioid receptor agonist, is not regulated by guanine nucleotides and sodium. AB - BW 373U86 is a novel, non-peptidic delta-opioid receptor ligand with agonist properties in mouse brain and in the mouse isolated vas deferens. The sensitivity of BW 373U86, and of the peptide delta-opioid agonists [D-Pen2,D-Pen5]enkephalin (DPDPE) and [D-Ala2,Glu4]deltorphin, to regulation by guanine nucleotides and sodium was evaluated in competition studies against the 5 selective radioligand [3H]naltrindole. The IC50 values for DPDPE and [D-Ala2,Glu4]deltorphin were significantly increased in brain and mouse vas deferens in the presence of Gpp(NH)p and NaCl. In contrast, the IC50 values for BW 373U86 were not altered in the presence of Gpp(NH)p and NaCl in either tissue. The data indicate that the agonist properties of BW 373U86 may not be affected by the supposed uncoupling of the alpha-subunit of the G-protein from a receptor thought to be G-protein linked. PMID- 8223953 TI - Gelsolin-actin complex is target for ADP-ribosylation by Clostridium botulinum C2 toxin in intact human neutrophils. AB - The gelsolin-actin complex was ADP-ribosylated by Clostridium botulinum C2 toxin and Clostridium perfringens iota toxin in lysates of human platelets and human neutrophils. When [32P]orthophosphate-labelled human neutrophils were treated with C. botulinum C2 toxin, [32P]ADP-ribosylated gelsolin-actin was precipitated with anti-gelsolin antibody. Stimulation of neutrophils by formyl-methionine leucine-phenylalanine decreased the interaction of gelsolin with ADP-ribosylated actin in toxin-treated cells. The data indicate that the gelsolin-actin complex is a pathophysiological substrate for actin-ADP-ribosylating toxins. PMID- 8223954 TI - Cocaine induces rapid loss of intracellular free Mg2+ in cerebral vascular smooth muscle cells. AB - Acute exposure of cultured canine cerebral vascular smooth muscle cells to low concentrations of cocaine HCl (10(-9) to (10(-7) M) resulted in significant, rapid (1 min) loss of intracellular free Mg ions ([Mg2+]i); these reductions (12 25%) in [Mg2+]i were reversible upon exposure to normal, Mg(2+)-containing physiological salt solution. These findings help to provide a rational basis for why cocaine can result in cerebrovasospasm and stroke. PMID- 8223955 TI - Effects of ethanol on contractile response of gall bladder isolated from guinea pig. AB - The effects of ethanol treatment in vitro and in vivo on gall bladder contraction were investigated using gall bladder strips isolated from guinea pigs. In vitro pretreatment of the strips with ethanol at a concentration of over 50 mM significantly attenuated the reactivity and sensitivity of contractile responses to KCl, acetylcholine and histamine in a concentration-dependent manner. Indomethacin treatment or removal of extracellular calcium remarkably reduced gall bladder contractile response to acetylcholine. The depressive effect of ethanol in vitro on gall bladder contraction was also noted in the presence of indomethacin or absence of calcium in the medium. The concentration-response curve of calcium-induced contraction in 40 mM KCl-depolarized gall bladder strip shifted to the right on pretreatment with ethanol. In the case of strips following the chronic administration of 3% ethanol solution ad libitum for 4 weeks, contractile responses to KCl, acetylcholine and histamine did not differ, compared to those in the pair-fed group. This chronic ethanol administration induced tolerance to the acute inhibitory effect of ethanol on gall bladder contractile responses to the agonists. Ethanol is thus shown to exert direct inhibitory action on gall bladder contraction by lowering the calcium sensitivity of the contractile apparatus of smooth muscle; it is unlikely that ethanol consumption would affect gall bladder motility in vivo, owing to the tolerance produced toward the acute inhibitory action of ethanol. PMID- 8223956 TI - The effects of colchicine or vinblastine on the blood calcium level in rats. AB - In order to clarify how microtubule inhibitors induce hypocalcemia, rats were injected with intravenous colchicine (1 mg/kg) or vinblastine (2 mg/kg). The blood calcium levels decreased rapidly, and the minimum values, reached 4 h after the injection, were 7.55 +/- 0.70 mg/100 ml (mean +/- S.D., P < 0.001) for colchicine and 7.61 +/- 0.17 mg/100 ml (P < 0.001) for vinblastine. At 24 h, these values returned to the normal range (10.13 +/- 0.42 mg/100 ml). The blood calcium values in rats fed a low calcium diet and in thyroparathyroidectomized rats were also reduced by colchicine. The incorporation of blood 45Ca into bone was reduced by the injection of colchicine. Histologically, the bone cells of rats injected with either drug were severely damaged 8 h after the injection. These results indicate that hypocalcemia may be mediated by interference with the regulatory mechanisms of bone cell calcium homeostasis, and that the destruction of microtubules may be closely related to the development of the hypocalcemia. PMID- 8223957 TI - Toxic injury to rat gut musculature following intraperitoneal administration of 2 t-butyl-4-methoxyphenol. AB - The 100-fold increase in toxicity of intraperitoneal (i.p.) rather than orally administered 2-t-butyl-4-methoxyphenol (BHA) is adduced to the depressive effect which this compound exerts on the contractility of the gut musculature. A structure/activity relation study shows the t-butyl group on the benzene ring as being the major determinant of i.p. BHA toxicity. Contractile activity, elicited by field electrical stimulation, acetylcholine or Ba2+, of the ileum longitudinal muscle preparation from BHA-treated rats was greatly reduced 30 min after i.p. injection, and almost absent during the subsequent 48 h. Electron-microscope examination of ileum longitudinal muscle also showed partial destruction of cell membranes 4 h after BHA administration with subsequent mitochondrial swelling and destruction of cristae, myofibrillar fragmentation and cell necrosis. Comparable suppression of contractile activity and morphological damage were observed in BHA or t-butylbenzene incubated ileum segments where longitudinal smooth muscle contractility was irreversibly depressed in a time- and dose-dependent manner. These convergent findings point to the toxic effect of i.p. BHA on gut musculature with consequent impairment of intestinal transit. PMID- 8223958 TI - Involvement of free radicals in cisplatin-induced emesis in Suncus murinus. AB - The participation of free radicals in cisplatin-induced emesis was investigated in the house musk shrew, Suncus murinus. Thiobarbituric acid (TBA) values, which indicate the degree of lipid peroxidation, in brain, liver and small intestine were increased significantly 60 min after the treatment with cisplatin (20 mg/kg, i.p.). Moreover, cisplatin (20 mg/kg, i.p.)-induced emesis was prevented by intraperitoneal injection of N-(2-mercaptopropionyl)glycine (MPG), a radical scavenging agent, with ID50 value of 130 mg/kg. However, MPG did not block the emesis induced by copper sulfate (40 mg/kg, p.o.), veratrine (0.5 mg/kg, s.c.) or serotonin (10 mg/kg, i.p.). We also investigated the effects of superoxide dismutase conjugated to polyethylene glycol and catalase, but the number of vomiting episodes and latency did not change significantly when these agents were intraperitoneally injected 30 min prior to or 20 min after the administration of cisplatin. MPG did not affect the antitumor effect of cisplatin tested in vitro. These results suggest that free radicals mediate emesis caused by cisplatin and that radical scavengers may become a new class of prophylactic drug against cancer-chemotherapeutic drug-induced emesis. PMID- 8223959 TI - Action of robenidine on the intestinal transport and digestion of nutrients in rabbit. AB - Robenidine is an anticoccidial guanidine used as an additive in rabbit fodder. Because its action is restricted to the small intestine, the present work addresses the question whether robenidine affects the growth of the animals, sugar and amino acid intestinal transport and membrane-bound intestinal digestion. For this purpose we have determined the intestinal transport of the substrates, and the enzymatic activity of neutral aminopeptidase and sucrase. We have found that robenidine diminishes the tissue accumulation of L-leucine and D galactose at long incubation times, and increases the transepithelial mucosal to serosal flux of both substrates. These results suggest that robenidine may stimulate the enterocyte basolateral membrane flux of sugars and neutral amino acids. These results have been corroborated by means of isolated brush border and basolateral membrane vesicles. Apart from these effects, robenidine has also been shown to increase the enzymatic activity of neutral aminopeptidase and sucrase and thus resulting in a better digestion of nutrients. PMID- 8223960 TI - Nitrate tolerance and aging in isolated rat aorta. AB - The purpose of this study was to determine whether there are age-related changes in the extent of in vitro-induced nitrate tolerance. Nitroglycerin pre-exposure (10 microM for 30 min) provoked a significant shift to the right of the dose response curve to nitroglycerin in aortae isolated from rats of 8 weeks, 12 and 18 months. However, this shift to the right was significantly larger at 18 months, both when KCl or phenylephrine were used as contractile agents. On the contrary, nitroglycerin pre-exposure did not significantly alter the dose dependent relaxation to Sin-1 (3-morpholinosydnonimine, the active metabolite of molsidomine) at 8 weeks, 12 and 18 months. These data indicate that the extent of the in vitro-induced nitrate tolerance is larger when aortae are isolated from senescent rats. This increase in tolerance does not appear to involve desensitization of guanylate cyclase. PMID- 8223961 TI - Inhibition by human leukocyte elastase of neutrophil-mediated platelet activation. AB - When human polymorphonuclear neutrophils and platelets were incubated with human leukocyte elastase before N-formyl-Met-Leu-Phe (FMLP) challenge, a time- and concentration-dependent inhibition of the resulting platelet activation was observed. Thus, when the mixed cell suspension was preincubated for 6 min with 1 microM elastase before stimulation of neutrophils with 0.5 microM FMLP, resulting aggregations and serotonin releases were respectively only 4.4 +/- 4.1% (n = 4) and 1.6 +/- 2.4% (n = 4) as compared to 41.6 +/- 5.2% (n = 9) and 71.3 +/- 16.0 (n = 9) for controls. A direct inhibitory action of elastase on neutrophil activation was ruled out, as well as a breakdown of cathepsin G, a mediator involved in neutrophil-mediated platelet activation. In fact, we demonstrated that the target for the inhibitory effect of elastase in such a cell-to-cell cooperation system was the platelet. This phenomenon is likely to play a role under in vivo conditions in pathologies in which a significant granulocytic proteolytic activity has been detected in the plasma. PMID- 8223962 TI - Inhibition of 5-hydroxytryptamine- and enterotoxin-induced fluid secretion by 5 HT receptor antagonists in the rat jejunum. AB - The effects of cholera toxin and heat stable Escherichia coli (E. coli) enterotoxin on intestinal fluid secretion are commonly considered to be mediated by cyclic nucleotides. It was demonstrated recently, by using the 5 hydroxytryptamine (5-HT)2 receptor antagonist ketanserin and the 5-HT3 receptor antagonist tropisetron, that 5-HT acts as an important mediator in cholera toxin- and heat stable E. coli enterotoxin-induced fluid secretion. In the present investigation ketanserin and tropisetron were compared with the newer 5-HT3 receptor antagonists ondansetron and granisetron versus 5-HT-, cholera toxin- and heat stable E. coli enterotoxin-induced fluid secretion in the rat jejunum in vivo. Both ondansetron and granisetron dose-dependently inhibited 5-HT- and enterotoxin-induced fluid secretion. Ketanserin blocked 5-HT-induced fluid secretion, but only diminished enterotoxin-induced effects even at higher doses. Tropisetron inhibited 5-HT- and cholera toxin-induced effects at high dose but only diminished heat stable E. coli enterotoxin-induced effects. We conclude that 5-HT3 receptors, located on enterochromaffin cells and nervous structures, are more important in mediating fluid secretion than 5-HT2 receptors, located on the epithelial cells. PMID- 8223963 TI - Antiplatelet effects of a new de-N-acetyl-lyso-glycosphingolipid. AB - Gal beta 1-->3GalN beta 1-->4Gal(3<--2 alpha Neu)beta 1-->4Glc beta-->1Sph (WILD20), a new glycosphingolipid, a breakdown product of the monosialoganglioside GM1 obtained through alkaline hydrolysis, shows dose dependent platelet anti-aggregating properties in vitro and in vivo. This effect is agonist- and species-independent. The family of lysosphingolipids, to which the compound belongs, is present in platelets particularly after thrombin treatment. WILD20 antiplatelet effect is due to the interference with ADP or thrombin-induced aggregation, probably via phospholipase A2 (PLA2) blockade; the substance is also effective when arachidonic acid is used as an agonist. Serotonin blood levels are also reduced. The substance, orally active at dosages of 0.1-0.01 mg/kg as antiplatelets agent, prolonged bleeding time without interfering with the coagulative or fibrinolytic processes. PMID- 8223964 TI - Improvement by diazepam of neuromuscular transmission blocked by anticholinesterase agents in mouse diaphragms. AB - Effects of diazepam on the neuromuscular transmission blocked by neostigmine were studied in isolated mouse phrenic nerve-diaphragm preparations. Diazepam, in the absence of neostigmine, had no significant effect on the neuromuscular transmission at concentrations lower than 100 microM, except to enhance muscle contractility. Single and train pulses-evoked endplate potentials (e.p.p.s) and miniature e.p.p.s (m.e.p.p.s) were also unaffected. At concentrations of 175 microM or higher, diazepam caused an axonal conduction block. However, neostigmine-induced twitch potentiation, spontaneous fasciculation and tetanic fade were antagonized by diazepam at 3.5-35 microM. Diazepam did not decrease the amplitude of neostigmine-augmented e.p.p.s and m.e.p.p.s but slightly reduced their decay time. The incidence of regenerative depolarization of endplates induced by repetitive stimulation in the presence of neostigmine, was decreased from 92% to 35% junctions and the duration shortened from 650 ms to 230 ms. The amplitude of train e.p.p.s was increased. It is suggested that diazepam reverses the muscle paralysis induced by anticholinesterase agents by inhibiting the regenerative release of acetylcholine. PMID- 8223965 TI - Effect of pentobarbital on postischemic SCH 23390 and rolipram binding in gerbil brain. AB - We investigated the postischemic alterations in dopamine D1 receptor and Ca2+/calmodulin independent cyclic adenosine monophosphate (cyclic AMP) selective phosphodiesterase in gerbils and examined the effect of pentobarbital on these alterations. [3H]SCH 23390 and [3H]rolipram, respectively, were used to label dopamine D1 receptor and Ca2+/calmodulin independent cyclic-AMP selective phosphodiesterase. Transient cerebral ischemia was induced for 10 min, and pentobarbital (40 mg/kg) was administered intraperitoneally 30 min prior to ischemia. 5 h after ischemia, [3H]rolipram binding decreased significantly in the striatum and hippocampus, whereas no significant change was found in [3H]SCH 23390 binding. 7 days after ischemia, however, there was a marked reduction in both [3H]SCH 23390 and [3H]rolipram binding in the striatum and hippocampus, where histological neuronal damage was found. Pentobarbital significantly ameliorated postischemic decreases in [3H]rolipram binding both 5 h and 7 days after recirculation in most areas studied. Furthermore, this drug significantly prevented postischemic reduction in [3H]SCH 23390 binding (only) 7 days after ischemia. These results suggest that alteration of cyclic AMP selective phosphodiesterase is more sensitive at an earlier stage after ischemic insult than that of dopamine D1 receptors. Our results also demonstrate that pentobarbital reduces the alteration in [3H]SCH 23390 and [3H]rolipram binding after cerebral ischemia. PMID- 8223966 TI - Effects of different factors in lead- and cadmium-induced hypothermia in mice. AB - The aim of this study was to investigate the relationship between the hypothermic effect induced by lead or cadmium chloride treatments and the cerebral metal levels attained. Mice were injected intraperitoneally with different doses of lead acetate or cadmium chloride at ambient temperatures of 22 degrees C and 35 degrees C, and rectal temperatures and brain metal levels were determined. At 22 degrees C, doses of lead acetate exceeding 25 mg/kg caused significant hypothermia and a rise of lead levels in the brain; this hypothermic effect was significantly inhibited at 35 degrees C. In mice repeatedly treated with 2 or 5 mg/kg lead acetate and exposed to 22 degrees C or 35 degrees C, no significant changes were observed in body temperature after the repeated exposure, although brain lead levels increased significantly. Cadmium chloride at 22 degrees C produced a significant fall in body temperature, and a rise in brain cadmium levels at doses of 2-4 mg/kg; the decrease in body temperature was partially inhibited at 35 degrees C. The results of the present study suggest that the hypothermic effect elicited by these heavy metals is related to both the level of metal absorbed by the brain and its rate of uptake. PMID- 8223967 TI - Endothelin-1 exacerbates focal cerebral ischemia without exerting neurotoxic action in vitro. AB - In a model of focal cerebral ischemia in mice, intracisternal injection of 5 and 10 pmol/mouse endothelin-1 significantly increased the infarcted surface area by 15.5% and by 23.5%, respectively. Endothelin-1 (0.01, 1, and 100 nmol/l) added to the primary neuronal cultures of chick embryo cerebral hemispheres for 1 h and 24 h did not influence the viability of the neurons or the protein content of the cultures. When applied simultaneously with 1 mmol/l sodium cyanide for 30 min, endothelin-1 (0.01, 1, and 100 nM) did not modify the hypoxia-induced changes. The results show that exogenously applied endothelin-1 could exacerbate cerebral ischemia, probably due to its vasoconstrictive properties and not to a direct neurotoxic effect. PMID- 8223968 TI - The effect of a range of alcohols on the contraction of guinea-pig ventricular myocytes. AB - Previous results have shown that ethanol and some anaesthetics have a negative inotropic effect on the heart. This study was undertaken to investigate the influence of a range on n-alcohols (with chain lengths from 2 to 8) on contractility in guinea-pig ventricular myocytes. The results demonstrate that the negative inotropic action of alcohols increases dramatically as the chain length increases. The concentration required to reduce the magnitude of contraction to 50% of control (IC50) was 274 mM, 26 mM, 1.4 mM and 235 microM for ethanol, butanol, hexanol and octanol, respectively. The relationship between the logarithm of IC50 and chain length was linear for all the alcohols tested (up to a chain length of 8). PMID- 8223969 TI - Retrovirus-mediated stable expression of human CYP2A6 in mammalian cells. AB - To study the pharmacological and toxicological significance of the human cytochrome P450 isoform CYP2A6, we expressed it in mammalian cells by retrovirus mediated gene transfer. The LXSN vector and PA317 packaging cells were used to create amphotropic recombinant retroviruses containing CYP2A6 cDNA. NIH 3T3 and HeLa cells were infected with these retroviruses and cell clones expressing CYP2A6 were selected. The integration of the CYP2A6 construct was verified by PCR analysis and northern blot analysis showed that a 5 kb mRNA containing the CYP2A6 was present in the cells. The integrated cDNA directed the expression of catalytically active CYP2A6 enzyme which has remained stable over numerous cell passages. No oxidation of several other P450 substrates was detected. The promutagen aflatoxin B1 was metabolized to intermediates binding to the host cell genomic DNA by the 3T3 2A6 cells. These cell lines are thus well suited for the study of the catalytic profile and the biological consequences of promutagen activation by the human CYP2A6 isoform. PMID- 8223970 TI - The aetiological factors in Class III malocclusion. AB - This retrospective, cephalometric study was undertaken in an attempt to identify the aetiological features which may be associated with a Class III malocclusion in the growing child. Four-hundred-and-ninety-five lateral cephalometric radiographs (210 control and 285 Class III films) formed the basis of the study. All subjects were Caucasian: the material was divided into four age groups, and males and females were examined separately. Comparisons were made between control and Class III children in each of these eight subgroups. Class III children showed differences in facial morphology in all facial areas examined, when compared with their control peers. The cranial base angle was more acute, the maxilla shorter and more retrusive, whilst the mandible was longer and more prominent. This was due in part to a more ventral position of the mandibular articulation. The proclined upper incisors were as far forward within the face in the Class III group as in the controls, but the retroclined lower teeth were even more labially placed. The range of individual variation was wide and few Class III children exhibited all characteristic features. Both control and Class III male faces showed their greatest increments of growth between 14 and 17 years of age: in control females, facial growth had ceased by this time, but in the Class III group, development remained active. The Class III female face seemed to have a tendency towards horizontal development, whereas the males exhibited a more vertical growth pattern. PMID- 8223971 TI - Early treatment of Class III incisor relationship using the chincap appliance. AB - Twenty-three patients (10 male, 13 female), with Class III relationships of the incisor teeth at the mixed dentition stage, were treated with the chincap appliance associated, where appropriate, with an upper removable appliance to procline upper incisors and to free the occlusion. Cephalometric analyses at the beginning and at the end of active treatment were compared with those in a carefully matched untreated control group selected from the Belfast Growth Study. At the beginning of treatment patients differed significantly from the controls in the SNB was larger, ANB was negative, Wits' analysis was negative, and the intermaxillary angle significantly reduced. The overjet was reversed, the lower incisors were ahead of A-Po, and the upper lip was far behind the E line. Significant changes brought about by the treatment were improvement in the Wits' relationship (but there was no significant change in the ANB angle), increases in the intermaxillary angle and lower facial height, improvement in the overjet, proclination of upper incisors and retroclination of lower incisors, backward movement of the lower incisors in relation to A-Po, and backward movement of the lower lip. The efficacy of the chincap appliance can be attributed to retroclination of lower incisors and downward movement of the mandible which may improve the jaw relationship without affecting the ANB angle. There were marked changes in the lip posture. PMID- 8223972 TI - A comparison of the forces required to produce tooth movement in vitro using two self-ligating brackets and a pre-adjusted bracket employing two types of ligation. AB - Friction in fixed appliance systems has received considerable attention in the recent literature, although that attributable to the type of ligation used has not been fully investigated. This in vitro study of 0.022 x 0.028 inch slot Minitwin, Activa ('A' Company, San Diego, California, USA), and SPEED brackets (Strite Industries Ltd., Cambridge, Ontario, Canada), investigates friction in two forms of self-ligating brackets and in two methods of ligating Minitwin straight wire brackets with polyurethane elastomeric ligatures. The resistance to sliding of rectangular archwires through the ligated brackets was measured on a vertically mounted Instron testing machine. The stainless steel archwires used were straight lengths of 0.016" x 0.022", 0.017" x 0.025", 0.018" x 0.025", and 0.019" x 0.025" Nubryte Gold (GAC International Inc., Central Islip, New York, USA). The results showed a significant reduction (P < 0.01) in frictional resistance in the Activa brackets compared with SPEED brackets by a factor of approximately 15. When the SPEED brackets were compared to Minitwin brackets, the reduction in friction was by 50-70 per cent (P < 0.01). The placing of 'figure-of eight' elastomeric ties increased friction by a factor of 70-220 per cent compared to conventional elastomeric ties (P < 0.01) except for 0.016" x 0.022" archwires. The results indicate that self-ligating brackets require less force to produce tooth movement because they apply less frictional contact to the archwire than conventionally tied siamese brackets. PMID- 8223973 TI - Transcranial oblique lateral radiography to verify the position of the mandibular condyles with the use of functional appliances. AB - By means of standardized transcranial oblique lateral (TOL) radiographs, the position of the mandibular condyles in the glenoid fossae of 14 patients with Class II division 1 malocclusion was studied at the beginning of treatment with or without functional appliances placed in position, and also at the end of treatment. A significant increase in the distance between the upper point of the condyle and the glenoid fossa was found when the appliance was worn for the first time. This displacement was found to disappear at the end of the treatment even when the appliance was placed in position. PMID- 8223974 TI - Orthodontics in Europe 1992. AB - The removal of economic barriers in Europe in 1992, began a new era in history and will have profound effects on orthodontics throughout Europe. In order to get an estimate of the orthodontic scene in each European country a questionnaire was sent to a well known orthodontist who was asked to fill in the form. The questionnaire consisted of enquiries into four areas of orthodontics. The first dealt with orthodontic specialization in the country and inquired into the numbers of orthodontists, where they practised, how they trained, and whether there was a specialist register. The second part dealt with the orthodontic societies, how many were there, how many members, and the frequency of the meetings. The third area asked about orthodontic practice, dealing with case load, types of appliances used, and the cost of treatment. The last section dealt with the future of orthodontics in their particular country. This related to the demand for orthodontics, the need for orthodontists and the changing patterns of orthodontic practice over the next decade. Twenty-three of the 26 countries in Europe when the questionnaire was sent out responded although some were unable to answer all the questions because orthodontics was not recognized in their country. PMID- 8223975 TI - Specialist postgraduate orthodontic training: a comparison of two European universities. AB - The background to current specialist training programmes in orthodontics in the United Kingdom and France is discussed, and the examination structure in the two countries compared with reference to those pertaining in Glasgow and Strasbourg. A sample of 40 of the most recently treated cases submitted by students in each of these two Universities for their final examinations is compared revealing marked differences in case type and treatment philosophy. The differences are explained, to some extent, by the circumstances pertaining to the delivery of orthodontic care in the two countries. PMID- 8223976 TI - Force delivery from a transpalatal arch for the correction of unilateral first molar cross-bite. AB - The forces and moments delivered by prefabricated transpalatal arches of the makes Unitek, GAC, and Ormco were studied in laboratory experiments. The Unitek and GAC arches were made of steel, and the Ormco arch of beta-titanium alloy (TMA). Three types of activation were investigated: for bilateral expansion in a statically indeterminate system and for unilateral expansion in a statically determinate system with and without torque activation. In addition to the arch design, composition, and mode of activation, the influence of arch size and degree of activation were studied. It was found that activation for unilateral expansion with the inclusion of torque in the statically determinate system produced forces and moments suitable for the correction of a unilateral cross bite. PMID- 8223977 TI - The use of magnets in orthodontic therapy: panel discussion. 68th Congress of the European Orthodontic Society. PMID- 8223978 TI - Ectopic eruption of maxillary canines. PMID- 8223979 TI - Feeding status-related effects of 17 beta-estradiol on liver 3-hydroxy-3 methylglutaryl CoA reductase. AB - Total and expressed 3-hydroxy-3-methylglutaryl CoA reductase was determined in hepatic microsomes prepared from fasted and fed control rats and 17 beta estradiol-treated rats. Plasma, liver and microsome cholesterol levels were also measured. Fasting resulted in significantly lower total and expressed specific activity of HMG-CoA reductase such that the expressed/total activity ratio doubled regardless of the endocrine status of the animals. Rats treated for either 3 or 21 days with 50 micrograms 17 beta-estradiol and starved for 24 hours exhibited greater reductase activity than untreated animals, the percentage of enzyme in effective form being maintained. When rats were fed on a commercial diet or a 4% cholestyramine-diet for five days, thus presenting higher basal HMG CoA reductase levels, the stimulatory effect of 17 beta-estradiol was not statistically significant. In contrast, when total reductase was reduced by dietary cholesterol, the estrogen effect was accentuated. Long-term estradiol treatment was associated with increases in liver weight, decreases in free and total plasma cholesterol, and microsome cholesteryl ester accumulation; while short-term estradiol treatment increased plasma esterified and total cholesterol as well as the microsomal content of cholesteryl esters. These findings suggest (i) that the two 17 beta-estradiol treatments employed in this study are associated with higher HMG-CoA reductase levels and this is particularly apparent when cholesterol synthesis occurs at a low rate and (ii) that plasma cholesterol levels are not strictly correlated with hepatic HMG-CoA reductase activity. PMID- 8223980 TI - Incidentally discovered adrenal masses: a functional and morphological study. AB - In the last two years we have examined 17 consecutive patients (11 females and 6 males, 20-66 years old) in whom an unsuspected adrenal mass was discovered by ultrasonography or computed tomography performed for unrelated reasons. Pathological diagnosis was available in 11 cases based on surgical excision in 9 (2 pheochromocytomas of 5 and 12 cm in diameter; 2 ganglioneuromas of 5 and 6 cm; and 5 benign cortical adenomas between 3 and 5 cm), autopsy in 1 (a disseminated malignant pheochromocytoma of 16 cm) and fine-needle biopsy in 1 (a pseudo adrenal mass of 6 cm, that was a regenerative hepatic nodule). The remaining 6 non histologically diagnosed masses were less than 3 cm in diameter. Endocrine studies showed elevated urinary excretion of catecholamines, vanillylmandelic acid and metanephrines in the pheochromocytomas and borderline high values in ganglioneuromas. A low plasma renin activity was encountered in 2 operated cortical adenomas and 3 non operated incidentalomas. In 2 of the latters aldosterone serum levels were elevated and the final diagnoses respectively were Conn's adenoma and dexamethasone-suppressible hyperaldosteronism with bilateral nodular hyperplasia. An inappropriate cortisol secretion was documented in a cortical adenoma removed. Radio-cholesterol scintiscan showed unilateral or increased uptake on the side of adrenal mass (concordant uptake) in the 5 benign cortical adenomas removed and in 4 non operated incidentalomas. A decreased uptake on the side of the adrenal mass (discordant uptake) was found in the 2 ganglioneuromas while an indeterminate bilateral uptake was found in the 2 remaining non operated incidentalomas and in the pseudo-adrenal mass.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223981 TI - Plasma aldosterone response to metoclopramide in patients with Cushing's disease. AB - The acute effect of metoclopramide on aldosterone and prolactin levels were studied in 8 control subjects and 11 patients with Cushing's disease. Metoclopramide (10 mg i.v.) induced a similar plasma prolactin response in control subjects and patients. No difference was found in basal aldosterone levels between the two groups. Apart from a more delayed aldosterone response to metoclopramide in the Cushing's disease group, no significant difference in the peak aldosterone values was found between the patients and controls. The short term treatment with DA2-receptor agonist bromocriptine was without effect on aldosterone response to metoclopramide in the Cushing's disease group. These results argue against an altered dopaminergic control of aldosterone secretion in patients with Cushing's disease. The lack of a bromocriptine effect on aldosterone response to metoclopramide is in accordance with the concept that the dopamine receptors involved in aldosterone regulation differ from classical DA2 receptors. PMID- 8223982 TI - Measurement of insulin-like growth factor I (IGF-I) in normal adults, patients with liver cirrhosis and acromegaly: experience with a new competitive enzyme immunoassay. AB - A competitive enzyme immunoassay for the determination of human insulin-like growth factor I in microtiter plates was established. Using a polyclonal antiserum raised in rabbits against hIGF-I ovalbumin conjugate the assay system was able to detect IGF-I at a range of 12-800 pg/well with a sensitivity of 10 pg/well. It showed a low (< 0.5%) cross reactivity with hIGF-II. The serum concentrations of IGF-I found by EIA agreed well with those found in a conventional RIA (r = 0.965, p < 0.001). Effects of age and sex on IGF-I levels were studied in 260 normal adults. There was no evidence for sex differences but a steep decline of values from the third to the fourth and from the eight to the ninth decade, respectively. To asses the diagnostic capability of the IGF-I determination in liver cirrhosis, 71 sera of patients classified according to Child classes (A-C) were measured. Although significantly diminished concentrations were found in class B vs A and in class C vs B, the diagnostic sensitivity in cross-sectional examinations proved to be low (class A: 0.33, class B: 0.67). Only in the case of extensively destroyed liver parenchyma (Child C: 0.94) IGF-I was a good indicator of impaired hepatocellular capacity. In 29 patients with acromegaly serum IGF-I levels were investigated. All patients with active acromegaly showed increased IGF-I levels. In contrast, in inactive or weakly active acromegaly values were considerably lower.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8223983 TI - Cyclic AMP formation in rat bone and kidney cells is stimulated equally by parathyroid hormone-related protein (PTHrP) 1-34 and PTH 1-34. AB - Parathyroid hormone-related protein (PTHrP) plays a major role in the pathogenesis of humoral hypercalcemia of malignancy. It interacts with the PTH receptor and has therefore a nearly identical effect on bone cells as PTH. However, PTHrP is thought to be less potent than PTH in stimulating adenylate cyclase in canine renal membranes, leading to the hypothesis of a differential efficiency in signal transduction by PTHrP with respect to bone vs kidney. In a homologous model with intact osteoblast-like cells (UMR 106) and primary kidney cells, both from the rat, we have tested N-terminal peptide fragments, based on the rat amino acid sequence 1-34, of PTH and PTHrP. Compared with PTHrP(1-34), rat PTH(1-34) had a similar relative potency in bone cells (85%) and in kidney cells (140%) in its ability to stimulate adenylate cyclase. Human PTH(1-34) was 5.6- to 6.5-fold less potent than rat PTH(1-34) in both cell types. In human osteoblast-like cells (SaOS-2), rat and human PTH were essentially equally potent compared to PTHrP(1-34) (identical sequence in rat and human) in stimulating cAMP accumulation. In conclusion, our study revealed the equipotency of rat PTH(1-34) and PTHrP(1-34) in stimulating intracellular cAMP formation in a homologous system of rat bone and kidney cells. There seemed to be no unique signal transduction mechanism of PTHrP to the adenylate cyclase in rat kidney cells compared with bone cells. PMID- 8223984 TI - Aspartate-aminopeptidase activity during the estrous cycle and the pregnancy in rat brain and pituitary gland. AB - The present report describes the activity of aspartate aminopeptidase, in 9 brain areas and in the pituitary gland, during the estrous cycle and several stages of pregnancy of the rat. The quantitation of the enzyme activity was performed by measuring the rate of hydrolysis of the chromogenic substrate Asp-beta naphthylamide. Aspartate aminopeptidase activity, in the pituitary gland and in several limbic areas, was found to be greater in cyclic than in pregnant rats. However the enzyme activity did not change significantly during either the estrous cycle and pregnancy. PMID- 8223985 TI - Neurosteroids do not regulate proopiomelanocortin-gene expression in pituitary cells. AB - Brain-derived steroids (neurosteroids) constitute a new group of steroids synthesized in brain cells independently from peripheral sources. While the membraneous actions of these steroids have been characterized in some detail, their genomic effects are less well studied up to now. In the present study, the action of the neurosteroids dehydroepiandrosterone, pregnenolone and pregnanolone on proopiomelanocortin (POMC)-gene expression was investigated in pituitary AtT 20 cells, which were cultured under serum-free conditions and at strictly controlled steroid levels. In these cells, POMC-gene expression is known to be negatively regulated by adrenocortical glucocorticoids. Quantitative Northern blot analysis showed that none of these neurosteroids or their conjugates affected POMC-mRNA levels even at the highest concentrations employed. The 21-OH steroid corticosterone was used as a control and markedly suppressed the synthesis of POMC-mRNA. These results suggest that neurosteroids do not control POMC-gene expression at the level of the pituitary corticotroph and exclude significant effects of these compounds at the glucocorticoid receptor. PMID- 8223986 TI - Experimental "chronic" African trypanosomiasis: endocrine dysfunctions generated by parasitic components released during the tryptanolytic phase in rats. AB - The disorders of the gonadotropic axis have been studied during the course of a "chronic" african trypanosomiasis induced experimentally in rats inoculated by the variant Trypanosoma brucei brucei AnTat 1.1.E. The levels of serum and pituitary LH as well as serum testosterone and corticosterone have been determined, during the infestation, at a particular period of the circadian cycle, in regard to the parasitemia variations. In addition, the inoculation of trypanosomal component fractions [obtained by concanavalin-A sepharose chromatography (conA-components)], has been performed in an attempt to define more exactly the nature of factor(s) producing the hypotestosteronemia in rats. This work evidenced that the hormonal parameter levels were predominantly decreased at the trypanolytic phase during the evolution of the disease. The action towards the hypothalamo-pituitary gonadal axis was attributed not only to peculiar trypanosomal enzyme(s) [a serine, thiol-dependent, cation sensitive endoprotease with a post-proline cleaving activity (purified from unretained conA fraction)], but also to protein and/or glycoprotein factor(s) released by the trypanosomes (components with affinity to the lectin). PMID- 8223987 TI - Myocardial atrial natriuretic factor gene expression after coronary ligation in rats. AB - The atrial natriuretic factor (ANF) messenger RNA (mRNA) content was determined in a rat model of acute myocardial infarction. Non-infarcted tissue of left and right ventricles were examined soon after ligation of the left descending anterior coronary artery. In the left ventricles, ANF mRNA/alpha-actin mRNA ratio increased to reach a maximum on day 7 and was sustained while in the right ventricle, the maximum was reached on day 1. The plasma ANF was significantly (p < 0.01) elevated from days 1 to 28 after coronary ligation. The primary compensatory response of ANF within the first 7 days may be attributed not only its synthesis in the left ventricle but also to its generation in the right ventricle in acute heart failure. PMID- 8223988 TI - Relationship between the monoamine and gonadotropin content in follicular fluid of preovulatory graafian follicles after superovulation treatment. AB - Noradrenaline (NA), serotonin (5HT), dopamine (DA), FSH, LH and prolactin (PRL) content was determined in 104 preovulatory follicular fluids obtained from 44 patients undergoing in vitro fertilization. The patients were given human menopausal gonadotropin (HMG) for ovarium stimulation, ovulation was induced with 10000 IU human chorionic gonadotropin (HCG) 34-36 hours prior to the follicular aspiration by vaginal ultrasound. Classification of the oocytes was performed by direct microscopic evaluation differentiating three groups of oocytes: Group I.: prophase I; Group II.: metaphase I; Group III.: metaphase II. There was no significant difference in monoamine and FSH content of follicular fluid at different stage of the oocyte maturation. LH and PRL significantly increased parallel with oocyte maturation (38.9; 48.8; 56.7 IU/l and 1324; 2382; 3134 IU/l). significant negative correlation was observed in Group I. between 5HT-LH (r = -0.64); in Group II. between NA-LH (r = -0.62) and NA-PRL (r = -0.51). Significant positive correlation were found in Group I. between FSH-LH (r = 0.63), in Group II. between LH-PRL (r = 0.56), in Group III. between NA-5HT (r = 0.66), NA-DA (r = 0.80) and 5HT-DA (r = 0.66). These observations suggest that action of LH and PRL may be negatively modulated by 5HT and NA in the final stage of oocyte maturation. PMID- 8223989 TI - Charcoal stripping augments type II oestradiol binding in cytoplasmic fraction of rat hypothalami. AB - Type II [3H]oestradiol binding was assessed by charcoal method in the low speed cytoplasmic fraction of intact female rat hypothalami. Augmented binding was observed after charcoal pretreatment applied prior to binding assay. On the other hand, loss of low affinity oestradiol binding was demonstrated after addition of protein-free ultrafiltrates of the cytosol from hypothalami or uteri. The presence of a cytosolic inhibitor activity specific for type II oestradiol binding in hypothalami is considered. PMID- 8223990 TI - BGP serum levels in patients with thyroid carcinoma before and 4-14 weeks after radioiodine therapy. AB - In the present short paper we compared the BGP and AP levels of 26 patients with thyroid carcinoma during radio-iodine therapy. BGP values reflect the bone turnover before and after radio-iodine therapy. There were no significant AP changes during this period. These results suggest that thyroid hormone has a direct effect on osteoblast function and that this effect can be best monitored by determination of BGP levels. PMID- 8223991 TI - Neonatal stimulation of beta-cells reduces the incidence and delays the onset of diabetes in a barrier-protected breeding colony of BB rats. AB - The off-spring in a barrier maintained colony of spontaneously type 1 diabetic BB/Wor/Mol-BB rats was treated with a daily injection of either saline, forskolin, arginine, glucose or both glucose and arginine for the first six days after birth. The incidence was reduced from 88% to 72% by the neonatal stimulation with arginine and glucose in combination, which also delayed the onset time from 76.0 +/- 2.2 days to 88.1 +/- 2.3 days. No such effect was observed after stimulation with either one of the compounds. Neonatal stimulation with forskolin also delayed the onset to 87.9 +/- 3.8 days, however without reducing the incidence. A higher incidence of diabetes was observed in the barrier-protected rats taking part in this study than in an earlier study using BB rats of the same origin, but kept under conventional conditions. PMID- 8223992 TI - A case of lipoatrophy with human insulin-therapy. AB - A case of progressive insulin-induced lipoatrophy is reported in an insulin dependent diabetic patient having been treated exclusively with human insulin. The lipoatrophy was stopped after insulin therapy was changed from subcutaneous injections to continuous subcutaneous infusion with human insulin. PMID- 8223993 TI - Type II iodothyronine 5'-deiodinase in mouse brown adipose tissue is stimulated by a single injection of isoproterenol in an oil/water emulsion. AB - A single subcutaneous morning injection of the beta-adrenergic agonist isoproterenol (ISO) (10 mg/kg body weight) in an oil/water emulsion (70/30; v/v) caused a marked increase in the activity of the enzyme type II iodothyronine 5' deiodinase (5'-D II) in the interscapular brown adipose tissue of BDF-1 mice. After a delay of 4 hours, the 5'-D II activity began to rise in an almost linear fashion and was increased 3-fold after 8 hours, when compared to the control values. The results indicate that this method of ISO administration may be a valid tool for artificial stimulation of brown adipose tissue in animals by beta adrenergic agonists. PMID- 8223994 TI - Hormonal control of glycogenolysis in isolated chick embryo hepatocytes. AB - Hepatocytes were isolated from 15-, 16-, 17-, and 18-day-old chick embryos. Glucagon, adrenaline, and dibutyryl cAMP (Bt2cAMP), individually or in combination, activated glycogenolysis in the isolated hepatocytes. The alpha adrenergic agonist phenylephrine did not increase glycogen breakdown. The action of adrenaline was abolished upon treatment of hepatocytes with a combination of the hormone and propranolol, a beta-adrenergic blocker. The effects of glucagon, adrenaline, and dibutyryl cAMP on glycogenolysis were not additive. Either hormone induced an increase in the concentration of cAMP. The activities of dephosphophosphorylase kinase and phosphorylase a were stimulated by each hormone or Bt2cAMP. It appears likely that glucagon and adrenaline serve as physiological regulators of hepatic glycogen breakdown during embryogenesis in chickens. PMID- 8223995 TI - Neural crest cell interaction with type VI collagen is mediated by multiple cooperative binding sites within triple-helix and globular domains. AB - Collagen type VI (Col VI) is a primary constituent of the extracellular matrix encountered by migrating avian neural crest cells in situ and is effective in promoting attachment and motility of these cells in vitro. In this study, we have explored the molecular mechanisms of neural crest-Col VI interaction by using quantitative assays for cell attachment and migration in vitro, proteolytic fragments of the collagen, and a panel of domain-specific monoclonal antibodies. Removal of the predominant portion of the amino-terminal globular domains of Col VI tetramers by pepsin digestion (P6 fragment) resulted in a > fivefold decrease in their cell adhesion and motility-promoting activity. Further digestion of P6 with bacterial collagenase, which causes a complete loss of the amino-terminal domains plus an adjacent triple-helical segment, did not affect adhesion but reduced migration down to 40% of that seen on undigested P6. Untreated and pepsin digested Col VI monomers were significantly less effective than their tetrameric counterparts and a M(r) 200,000 fragment, generated from pepsin-digested monomers by a second pepsin treatment, only retained 40% of the motility-promoting activity while preserving the adhesive capacity. A mixture of amino- and carboxyl terminal globular domains supported both cell attachment and migration. While neural crest cells adhered equally well to the individual intact alpha 1 (VI)/alpha 2(VI) and alpha 3(VI) chains, they migrated most extensively on the alpha 3(VI) chain. Conversely, pepsin-digested individual alpha chains were significantly less effective in promoting cell adhesion and locomotion. Selective preincubation of Col VI microfilaments and isolated tetramers with a panel of monoclonal antibodies against triple helix, carboxyl-terminal, and amino-terminal epitopes of the different constituent chains differentially perturbed neural crest cell attachment and migration. Sites differentially involved in neural crest cell attachment and migration seemed to be present at the carboxyl termini of the alpha 1(VI) and alpha 2(VI) chains and at the amino-terminus of the alpha 3(VI) chain. The results suggest that neural crest cells interact with Col VI through multiple and cooperative binding sites present within its triple-helical and globular domains. The differential involvement and efficiency of these sites in stimulating neural crest cell adhesion and migration is strongly determined by the supramolecular organization of the collagen and requires inter- and intramolecular structural integrity. Since neural crest cell attachment and migration on Col VI was completely inhibited by anti-beta 1 integrin antibodies, there is evidence that this class of integrins is essential for the neural crest cell--Col VI interaction. PMID- 8223996 TI - Effect of donor age on epidermal growth factor processing in man. AB - Epidermal growth factor (EGF) is a well-characterized mitogen whose effectiveness decreases with age both in vivo and in vitro. Previous studies utilizing the Hayflick aging model of early versus late-passage fibroblasts failed to demonstrate changes that might account for the unresponsiveness. In contrast, we now report striking differences in EGF receptor (EGFR) number, affinity, and rate of EGF/EGFR internalization in early-passage dermal fibroblasts derived from newborn versus young adult versus old adult donors. These data demonstrate critical differences between the two models of cellular aging, provide an explanation for the age-associated loss of EGF responsiveness, and may explain in part the tendency toward impaired wound healing in the elderly. PMID- 8223997 TI - Lysosomal enzyme transfer from different types of lymphoid cell. AB - The direct transfer of certain lysosomal enzymes during cell-to-cell contact between normal lymphocytes and enzyme-deficient recipient cells has previously been reported in vitro and may play an important role in the correction of lysosomal storage diseases by bone marrow transplantation in vivo. In the present study we have used a number of different T, B, and plasma cell lines to examine the expression and immunological specificity of the transfer of the lysosomal enzyme, beta-glucuronidase (Gus). Each of these groups of cell had differing intracellular and secreted levels of Gus activity, which were nevertheless similar within each group. Dermal fibroblasts deficient in the Gus enzyme acquired substantial amounts of additional activity when they were cultured together with the T cells, the B cells, or the plasma cells. This occurred by the direct transfer of Gus from all three types of cell. In addition, with plasma cells, which had very high intracellular enzyme activity and also secreted high levels of Gus into their culture medium, the secreted enzyme was readily internalized by the fibroblasts via the mannose 6-phosphate receptor (MPR). It was notable that the purified endogenous enzymes from plasma cells as well as from B cells, but not from T cells, were also endocytosed by the fibroblasts utilizing this receptor-mediated process. Although the Gus activity from all the cell lines examined had the same molecular size, polyacrylamide electrophoresis and isoelectric focusing patterns showed that the immunologically distinct types of lymphoid cell have characteristic, unique pathways of post-translational lysosomal enzyme processing. These results show that the transfer of lysosomal enzymes from lymphoid cells can occur by two distinct mechanisms, both likely to have important roles in enzyme replacement therapy. PMID- 8223998 TI - The endoplasmic reticulum-sarcoplasmic reticulum connection. II. Postnatal differentiation of the sarcoplasmic reticulum in skeletal muscle fibers. AB - The postnatal differentiation of sarcoplasmic reticulum (SR) of rabbit skeletal muscles (the slow-twitch soleus and the fast-twitch adductor muscles) was monitored between Days 1 and 12 by following on Western blots the expression and accumulation of molecular markers specific not only for the muscle endomembrane system, i.e., calsequestrin (CS) and the ryanodine-sensitive Ca2+ release channel, but also for the endoplasmic reticulum (ER) at large, i.e., BiP, calnexin (CN) and calreticulin. Our results demonstrate that SR development, documented by the increase of the SR fractional volume, terminal cisternae proliferation, and reorientation of triads, is accompanied by the accumulation of the SR-specific proteins and also of CN, with no change of the other ER general markers. Moreover, the distribution of two of the markers, BiP and CS, was investigated by immunocytochemistry at both the light and the electron microscope level. At Day 1 CS was found to be concentrated both within the few recognizable triad terminal cisternae and within the lumen of numerous, apparently discrete cisternae and tubules, widely scattered throughout both the contractile and the subplasmalemmal areas of the cytoplasm. These structures remain evident until Day 12, when most triad junctions have acquired proper configuration, composition and orientation. BiP, on the other hand, appears widely distributed within the ER/SR of the fibers. From the early stages of postnatal development it does colocalize with the Ca2+ binding protein in the lumen of the CS-rich structures and appears also within the longitudinal SR and the conventional ER cisternae. PMID- 8223999 TI - Chromatin bridges between sister chromatids induced in late G2 mitosis in CHO cells by trimethylpsoralen + UVA. AB - A hypothesis is proposed that sister chromatid pairing is due, at least in part, to the pairing between DNA strands belonging to each of the two sister chromatids (conservative pairing). To test this hypothesis interstrand DNA cross-links were induced in late G2-mitosis in CHO cells in order to bind covalently paired DNA strands eventually coming from both sister chromatids and detect the consequent chromatin bridges between sister chromatids (SCCBs). Therefore cells were treated with trimethylpsoralen (TMP) + UVA (365 or 405 nm). Chromatin bridges in ana telophase were induced by an UVA irradiation at 365 nm, which gives rise to both monoadducts and cross-links, but not by a 405-nm irradiation, which gives rise only to monoadducts. An analysis of colchicine-induced c-anaphases demonstrated that such chromatin bridges were really SCCBs and that terminal regions of chromosomes were particularly involved. The evolution of SCCBs was studied to rule out that they were masked isochromatid exchanges. So TMP + UVA-treated cells were induced to polyploidize with colchicine and labeled with 5 bromodeoxyuridine. Cells treated with TMP + UVA in G2-mitosis appeared as M1 tetraploid c-metaphases; in such cell populations there was not an increase in isodicentric chromosomes, which are derived from isochromatid exchanges. The present data, as a whole, support the hypothesis that a "conservative pairing" between DNA strands of sister chromatids can be present in mitosis. PMID- 8224000 TI - The comparison of DNA synthesis inducibility by high potassium treatment in various nonproliferating cells. AB - Previous studies have demonstrated that treatment of fibroblasts from confluent, density-inhibited cultures with 50 mM KCl solution led to the onset of DNA replication. In the present study we show that such treatment can induce aphidicolin-sensitive DNA replication in differentiated nondividing cells: dorsal root ganglia neurons and in vitro differentiated myotubes formed by L6 myoblasts. Murine peritoneal macrophages and macrophages/granulocytes derived from human promyelocytic leukemia cells HL60 are refractory to high potassium treatment. These results confirm the nonuniformity of the nonproliferative state of various differentiated cells. PMID- 8224001 TI - Plasma membrane cholesterol is utilized as steroidogenic substrate in Y-1 mouse adrenal tumor cells and normal sheep adrenal cells. AB - Previous studies from this laboratory indicate that plasma membrane cholesterol acts as an important source of steroidogenic substrate for MA-10 Leydig tumor cells. The present studies were designed to generalize these findings to other steroidogenic cells and to another species. Studies were performed using the Y-1 murine adrenal tumor cell line and primary cultures of sheep adrenocortical cells. Treating Y-1 cells with the acyl coenzyme A:cholesterol acyltransferase inhibitor, 58-035, caused cellular cholesteryl ester depletion and rendered more apparent the effect of dibutyryl-cAMP to cause cellular free cholesterol depletion. Radioactive 20 alpha-dihydroprogesterone was synthesized by Y-1 cells that had been plasma membrane-labeled with [3H]-cholesterol. Primary sheep adrenal cultures that had been cholesteryl ester-depleted also demonstrated cellular free cholesterol depletion after stimulation with dibutyryl cAMP. Plasma membrane label was converted to steroid hormones in these cells as well. Taken together, these data indicate that the use of plasma cholesterol is not restricted to the MA-10 cells. The present data indicate that both neoplastic mouse adrenal tumor cells and normal sheep adrenal cells utilize plasma membrane cholesterol. PMID- 8224002 TI - Significance of the extracellular domain and the carbohydrates of the human neutrophil N-formyl peptide chemotactic receptor for the signal transduction by the receptor. AB - The N-formyl peptide chemotactic receptor of human neutrophils possesses a 2-kDa papain-removable extracellular domain that contains two N-linked oligosaccharide side chains and is not required for the high-affinity ligand binding. In the present study, the significance of the extracellular domain and the carbohydrates for signal transduction was elucidated by measuring the N-formyl hexapeptide induced intracellular free calcium ([Ca2+]i) and the change in myeloperoxidase secretion in the control and papain-treated human neutrophils. [Ca2+]i was monitored both in cell suspension and individual cells with intracellularly trapped Fura 2 acetoxymethyl ester, using spectrofluorometric analysis and fluorescence ratio image analysis, respectively. The exposure of the cells in suspension to N-formyl hexapeptide resulted in an immediate, dose-dependent burst of elevated [Ca2+]i, which was virtually identical in both control and papain treated cells with respect to the extent and kinetics. The maximum burst was 1.6 fold and was obtained at 10(-6) M hexapeptide. The individual control and papain treated cells responded to 10(-6) M hexapeptide in a similar manner with several successive transients of [Ca2+]i, the maximum level being 3.0-3.5 microM. In both groups the [Ca2+]i transient began initially in the cell periphery, expanding rapidly throughout the cells. Concomitantly, the cells became polarized, and their chemokinesis increased. The secretion of myeloperoxidase was monitored as a physiological end response to N-formyl chemotactic peptides. The exposure of the control and papain-treated cells in suspension to hexapeptide resulted in a dose dependent secretion of myeloperoxidase. The maximum secretion after exposure to 10(-8)-10(-6) M hexapeptide was equal in control and papain-treated neutrophils. These results indicate that the functional properties of the membrane-inserted N formyl peptide chemotactic receptor are inherent to the receptor's transmembrane and intracellular domains, as far as binding of the ligand and subsequent receptor activation are concerned. PMID- 8224003 TI - Regulation of vitamin A transport into cultured stellate cells of rat liver: studies by anchored cell analysis and sorting system. AB - Stellate cells (SC) in the liver store the most retinoid in the body, but the mechanisms of specific retinoid transport into SC remain to be elucidated. In this study, to analyze the retinoid content of cultured SC, we employed an anchored cell analysis and sorting system (ACAS), which provides fluorescence analysis of single cultured cells under the phase-contrast microscope by utilizing a laser. First, we examined the effect of retinol binding protein (RBP) on retinol transport into cultured SC of rat liver. Rat holo-RBP added to the medium inhibited retinol uptake into SC. We also prepared RBP-free human serum by affinity chromatography using conjugated anti-human RBP IgG and compared retinoid fluorescence of SC cultured in human serum with or without RBP. No significant difference in retinoid fluorescence intensity was observed between SC cultured with and without holo-RBP. Second, the removal of cellular retinol by esterification may be important for the continued uptake of retinol. Retinyl esters are stored in lipid droplets of SC. Therefore we examined the relationship between the lipid droplet number and the retinoid fluorescence intensity in SC which were cultured in medium containing retinol for 1-3 days. The increases in lipid droplet number and in retinoid fluorescence in SC were almost parallel. Progesterone, previously shown to increase the esterification of retinol by lecithin:retinol acyltransferase (LRAT) in vitro, was added to the SC medium; progesterone facilitated retinol uptake in cultured SC. In conclusion, RBP did not facilitate specific retinol transport into SC. However, the specific transport of retinol is likely to be dependent on the intracellular esterification of retinol by LRAT in SC. PMID- 8224004 TI - Formation of ductular structures in vitro by rat pancreatic epithelial oval cells. AB - In the rat pancreas oval cells were shown to contain albumin transcripts and are considered to be precursors of pancreatic hepatocytes. Pancreatic oval cells have been maintained in our laboratory as a cell line. In the present study we have examined the differentiation potential of rat pancreatic epithelial oval cells under different culture conditions using light and electron microscopy and in situ hybridization techniques. Oval cells cultured on collagen gels formed a monolayer. However, when oval cells were embedded in collagen gels and cultured with or without fibroblasts, they readily organized into duct-like structures within 1 week. Over the next 3 weeks there was a gradual increase in the number and size of ductules. Morphological examination of duct-like structures showed true lumen formation. Ultrastructurally, cells showed features of ductal cells characterized by well-formed cell junctions, interdigitation of plasma membranes, well-developed cytoplasmic organelles, and apical microvilli. Light microscopic autoradiography showed proliferation of cells lining the ductules throughout the 4-week culture period. Albumin mRNA expression was demonstrable by in situ hybridization with albumin probe in oval cells cultured on collagen gels and embedded in collagen gels. Results of these studies indicate that under these in vitro conditions, oval cells appear to express albumin transcripts but exhibit duct-like morphogenetic features. PMID- 8224005 TI - The frequency of immortalization of human fibroblasts and mammary epithelial cells transfected with SV40 large T-antigen. AB - SV40 T-antigen-expressing human cells generally have an extension of lifespan until a period called "crisis" begins. On rare occasions a clone of cells emerges from the population in crisis and gives rise to an immortalized cell line. The present study compares the frequency of immortalization of cells from two different human lineages, lung fibroblasts and mammary epithelial cells. Most of the T-antigen-transfected clones from both cell types failed to immortalize, however, within those clones which were immortalization-competent the frequency of escape from crisis was found to be much higher (10(-5)) in mammary epithelial cells than in human fibroblasts (3 x 10(-7)). The frequency of escape from crisis in fibroblasts could be increased by chemical mutagenesis or by infection with a defective retrovirus. T-antigen-transfected fibroblasts were uniformly highly aneuploid both before and after crisis. In contrast, many SV40 T-antigen- and human papilloma virus 16 E6- or E6/E7-transfected mammary epithelial clones maintained a subpopulation of pseudodiploid cells prior to crisis, and the immortal cells that emerged following crisis were generally pseudodiploid. The higher frequency of escape from crisis in mammary epithelial cells is best explained by postulating a mutational inactivation of one allele of a critical gene followed by the elimination of the remaining wild-type alleles, with a much higher frequency of this second event in mammary epithelial cells due to their reduced ploidy compared to that in T-antigen-transfected fibroblasts. The results are discussed in terms of the regulation of telomerase activity and the M1/M2 model of cellular senescence. PMID- 8224006 TI - Mitogen-independent DNA synthesis by fetal rat hepatocytes in primary culture. AB - We have identified conditions under which late gestation fetal rat hepatocytes in primary culture proliferate in the absence of serum, polypeptide growth factors, or insulin. Fetal hepatocytes, cultured in defined minimal essential medium (MEM) with hydrocortisone, synthesized DNA within the first 6 h after plating and for up to 72 h. Rates of thymidine incorporation into DNA by fetal hepatocytes exceeded peak rates seen with adult rat hepatocytes. The latter were quiescent following isolation, with DNA synthesis only occurring after 48 h exposure to insulin plus epidermal growth factor. Although they exhibited a high rate of DNA synthesis, the fetal hepatocytes retained sensitivity to added mitogens; DNA synthesis was stimulated three- to fourfold by subnanomolar concentrations of TGF alpha. Fetal hepatocytes also were sensitive to the growth inhibitory effects of TGF-beta at concentrations below 10 pM. Finally, ontogenic changes in serum- and mitogen-independent fetal hepatocyte growth were observed, with declining rates of DNA synthesis as term approached. We speculate that the ability of fetal rat hepatocytes to synthesize DNA independent of added serum or mitogens may coincide with a proliferative in vivo phenotype. PMID- 8224007 TI - Insulin gene can be expressed in the absence of Isl-1. AB - The possible role of Isl-1 in insulin-gene transcription was investigated using Northern blot analysis to determine whether insulin and Isl-1 gene expression are correlated in various somatic cell hybrids. Among several hybrid cell lines obtained by fusing insulin-producing rat insulinoma (RIN) cells and mouse spleen cells, three (RR2, RR5, and RR11) had amounts of insulin transcripts similar to those of the parental RIN cells, although two of them, RR5 and RR11, lacked Isl-1 protein. In contrast, two RIN x mouse L cell hybrids where insulin expression was extinct still expressed Isl-1. RT-PCR analysis showed that Isl-1 transcripts are widely distributed in various mouse tissues, including pancreas, brain, lung, thymus, and ovary. These results indicate that Isl-1 is not essential for insulin gene expression. PMID- 8224008 TI - Primary culture of flow cytometry-sorted rat mammary epithelial cell (RMEC) subpopulations in a reconstituted basement membrane, Matrigel. AB - We studied the growth of rat mammary epithelial cell (RMEC) subpopulations cultured in Matrigel. Mammary glands were removed, minced, and enzymatically dispersed. Mammary organoids (ductal and endbud fragments) were collected, enzymatically monodispersed, filtered, and labeled with fluorescein isothiocyanate-peanut agglutinin (PNA) and phycoerythrin-anti-Thy-1.1 monoclonal antibody. By flow cytometry, we separated four RMEC subpopulations: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). The ultrastructural characteristics of sorted cells were examined by transmission electron microscopy. Thy-1.1+ cells appeared to be myoepithelial cells with actin filaments. PNA+, B-, and B+ cells were morphologically similar. Sorted RMEC subpopulations, unsorted mixed cells, and undispersed organoids were seeded in Matrigel and cultured in either a complete hormone medium (CHM) with 10% fetal bovine serum, progesterone, estradiol, hydrocortisone, insulin, and prolactin or a serum-free medium containing EGF, insulin, hydrocortisone, and human transferrin. Several types of colonies were observed: stellate, ductal, webbed, squamous, and lobuloductal colonies. At the immunocytochemical and electron micrograph levels, casein proteins were predominantly localized near the apical surfaces of the cells or in the lumina of ductal or lobuloductal colonies. Laminin and collagen IV were detected only in the basal lamina of ductal structures cultured in CHM. RMEC and subpopulations from immature virgin rats developed the capacity to synthesize and secrete lipid and casein when grown in Matrigel in serum medium or under defined serum-free conditions. PMID- 8224009 TI - The effect of three dimensional collagen type I preparation on the structural organization of guinea pig enteric ganglia in culture. AB - In vivo, cellular relationships in the myenteric plexus are characterized by unusual compactness and by the arrangement of neurons and glia into ganglia and interconnecting strands. These features are lost when the myenteric plexus is placed in culture. In the present paper we test whether collagen type I, a major component of the matrix that surrounds the plexus in vivo, might have a role in maintaining normal neuron-glia relationships in this system. We report that a three-dimensional gel of rat tail collagen prevented the disaggregation of the guinea pig myenteric plexus in culture and induced the formation of a compact plexus-like cellular network when applied to disaggregated plexus cultures. These effects were not observed with soluble collagen. Immunohistochemical evidence was also obtained for synthesis of type I collagen by enteric glia. These observations indicate that type I collagen in a three-dimensional organization is capable of inducing and maintaining both the unusual compact organization of neurons and glial cells within myenteric ganglia and also the characteristic organization of these cells into an orderly network of ganglia and interconnecting strands. PMID- 8224010 TI - Secreted collagen induced by ascorbic acid in L5 cloned muscle cultures does not affect acetylcholine receptor expression. AB - Previous studies have shown that ascorbic acid increases both the total surface acetylcholine receptor (AChR) expression and the mRNA for the alpha-subunit of this receptor in myotubes of cloned L5 muscle cultures. Since ascorbic acid increases collagen synthesis in fibroblasts, we studied the effect of ascorbic acid on collagen secretion in L5 muscle cells and investigated the possibility that the effects of ascorbic acid on collagen and AChR are related. We report that L5 muscle cells secrete collagen types I, III, and V, with collagen type I being the most abundant species, and that accumulation of secreted collagens increased in the medium approximately two- to ninefold within 3 h of ascorbic acid treatment. The increase in surface AChRs, on the other hand, developed more slowly, and was detected only about 20-24 h after ascorbic acid treatment. A short (5 h) treatment with ascorbic acid is, however, sufficient to trigger an increase in AChRs 24 h later. Since ascorbic acid caused a rapid increase in collagen secretion, whereas the effect on total surface AChRs occurs more slowly, we tested the possibility that an increase in secreted collagen might be necessary for the increase in AChRs. However, when the L5 cultures were treated with bacterial collagenase, the ascorbic acid-induced increase in secreted collagen was abolished but its inductive effect on AChRs was unchanged. The increase in secreted collagen is therefore not necessary for the increase in AChRs to occur. PMID- 8224011 TI - Changes in mitochondrial rhodamine-123-fluorescence populations of rat hepatocytes in primary culture. AB - Flow cytometry analysis of mitochondria isolated from rat hepatocytes at different stages of development revealed two different rhodamine-123-stained fluorescence populations distinguishable by their main fluorescence channel. The high-fluorescence population (HFP) was minor, accounting for about 32, 33, and 23% of the total mitochondrial fraction in hepatocytes from preterm, term, and early (1 h) newborn rats, respectively. The percentage of HFP decreased during the first 20 h of hepatocytes in culture from preterm and term fetuses but not those from early newborns, in which the decrease occurred during the second day of culture. However, after 20 h in culture more than 80% of hepatocytes reached the G0/G1 phase, whatever the stage of development in which they were sown. This suggests that the observed changes in the fluorescence populations are not associated with the cell cycle; instead these changes mimic those observed during postnatal development and are therefore presumably due to mitochondrial differentiation. PMID- 8224012 TI - Cytotoxic activity of rev protein of human immunodeficiency virus type 1 by nucleolar dysfunction. AB - The rev protein (Rev) of the human immunodeficiency virus type 1 (HIV-1) is known as a post-transcriptional regulator of viral gene expression. It is located in the cell nucleolus. Transiently expressed Rev caused nucleolar ballooning and deformity with aberrant accumulation of rRNAs, and de novo synthesis of rRNAs decreased dramatically in these cells. However, similarly expressed rex protein (Rex) of the human T-cell leukemia virus type I, which is a functional homologue to Rev, did not affect nucleolar structure and function. Rev expression resulted in cell death with nucleolar destruction in an inducible cell line. Analysis of Rev mutants revealed that both the nucleolar targeting signal of Rev and the multimerization domain are prerequisites to the nucleolar disintegration by Rev. Human T-cells acutely infected with HIV-1 contained nucleoli which were deformed and filled with Rev, but chronically infected cells had intact nucleoli. Involvement of Rev in cytopathic effects in HIV-1 infection is discussed. PMID- 8224013 TI - Suppression of phase separation in bovine gamma IV crystallin solutions: effect of modification by charged versus uncharged polar groups. AB - gamma IVa Crystallin, the ocular lens protein with the highest critical temperature (Tc) for phase separation, has been chemically modified with N ethylmaleimide (NEM) at neutral pH. NEM, a polar but uncharged modifier reacts with the cysteine residues of the protein. A maximum of 80-85% of the thiol groups are modified and the phase separation temperature is lowered by about 10 degrees C at a protein concentration of 30 mg ml-1. The coexistence curve of the NEM-modified protein with nearly four out of the six cysteines modified was determined and compared with that obtained by modifying the protein with a charged reagent, N-bromoacetylethanolamine phosphate (NBAEP). NBAEP modifies both cysteine and methionine residues of gamma IVa crystallin. The results of these two modification studies indicate that for the same total degree of modification of the sulfur-containing residues, the suppression in Tc due to the charged NBAEP is at least twice as large as that due to the polar but uncharged NEM. In order to obtain some measure of the relative hydrophilicities of the two modifiers, we have estimated the free energies of hydration of the nonionic segments of NEM and NBAEP, using structural additivity schemes. We find that the intrinsic hydrophilicities of the nonionic segments of these two modifiers are nearly equal. Hence the much larger suppression of Tc resulting from NBAEP modification is most probably due to the strongly polar, doubly charged phosphate group. PMID- 8224014 TI - Stage-specific binding of peanut agglutinin to aggregates of degenerating photoreceptor cells in the rd mouse retina. AB - Peanut agglutinin, a lectin with high binding affinity for galactose galactosamine disaccharide, was used to monitor changes in the photoreceptor cell layer of mice with inherited retinal degeneration. Mice homozygous for the retinal degeneration (rd) gene exhibit a rapid loss of rod photoreceptor cells in the first postnatal month. Previous studies have shown that aggregates of peanut agglutinin-binding cells are observed in the outer nuclear layer in the retinal degenerative mouse at between postnatal days 10 and 18, a period during which massive photoreceptor degeneration occurs in this mutant. This study was performed to determine whether these peanut agglutinin-positive cell clusters represent degenerating photoreceptor cells or, alternatively, macrophages that have migrated into the photoreceptor cell layer. Electron microscopic cytochemistry, using horseradish-peroxidase-conjugated peanut agglutinin, was used to trace cellular processes of peanut-agglutinin-stained cell clusters. Additionally, macrophage-specific antibodies were employed to determine whether macrophages were present in the clusters. The cell clusters did not react with macrophage-specific antibodies. However, processes of cells in peanut-agglutinin bound clusters could be traced by electron microscopic serial sections to both the outer limiting membrane and the outer synaptic layer. These results provide strong evidence that peanut-agglutinin-bound cells seen during this stage of degeneration in the rd mouse are degenerating photoreceptor cells. Since peanut agglutinin has been shown to bind preferentially to cone, but not to rod, photoreceptor cells, the results also suggest that the clusters may be aggregates of degenerating cones. PMID- 8224015 TI - Mechanisms of acetylcholine-induced relaxation in dog external and internal ophthalmic arteries. AB - Mechanisms underlying the relaxant response to acetylcholine (ACh) were examined in dog external (EOA) and internal ophthalmic arteries (IOA). Acetylcholine produced relaxation in EOA and IOA, partially contracted with prostaglandin (PG) F2 alpha. The relaxations induced by ACh in these arteries were not inhibited by endothelium denudation. The ACh (10(-8)-10(-5) M)-induced relaxations in EOA and IOA were abolished by treatment with atropine or indomethacin and markedly suppressed by tranylcypromine, a PGI2 synthesis inhibitor, but not influenced by hexamethonium. On the other hand, the relaxant response to the highest concentration used (10(-4) M) was partially attenuated but not abolished by treatment with atropine, indomethacin or hexamethonium. This relaxation under treatment with atropine was abolished by NG-nitro-L-arginine (L-NA), a nitric oxide synthase inhibitor, but not by indomethacin, whereas the response under treatment with hexamethonium was abolished by indomethacin, but not by L-NA. Combined treatment with indomethacin and L-NA or oxyhaemoglobin abolished the ACh (10(-4) M)-induced relaxation, or reversed it to a contraction. It may be concluded that relaxations induced by low concentrations of ACh in dog EOA and IOA are associated possibly with the release of PGI2 through activation of muscarinic receptors located in subendothelial tissues, including smooth muscle, and the relaxant response to the high concentration (10(-4) M) of ACh is mediated mainly by the release of nitric oxide through activation of nicotinic receptors in nitroxidergic nerves. PMID- 8224016 TI - ZO-1 in corneal epithelium; stratal distribution and synthesis induction by outer cell removal. AB - The tight junctions (TJs) present exclusively in between the superficial cells (SC) are an important component for the barrier and ion secretory functions of the corneal epithelium. Electrophysiological studies have demonstrated that digitonin-induced devitalization of overlying cells induces the de novo generation of a paracellular barrier between the intrastratal cells becoming exposed to the outer surface. It was also shown that the closer the exposed cells to the SC position the higher their competency for this activity. We have now examined the spatial and quantitative distribution of ZO-1, a protein closely associated with the cytosolic face of TJs in the rabbit. Immunohistology showed: (a) no detectable ZO-1 in basal cells; (b) incipient punctuate accumulations in the wing cells; (c) numerous foci and a diffuse cytosolic staining in the squamous (SQ) cells; and (d) strong staining at the apical TJ locations in the superficial SQ layer. Immunoblot analysis of separated layers showed a SQ:basal cell ZO-1 concentration ratio in excess of 100. Devitalization of the SQ layers induced strong ZO-1 synthesis in the newly exposed wing cells where electron microscopy showed time-dependent development of TJs. A similar ZO-1 increase was observed in the basal cells after removal of all suprabasal cells by a low [Ca2+] incubation method. These results provide a biochemical correlate to the previous electrophysiological findings and show that expression of ZO-1 in the corneal epithelium is intimately related to the development of the superficial cell phenotype. PMID- 8224017 TI - Effects of EGF, IL-1 and their combination on in vitro corneal epithelial wound closure and cell chemotaxis. AB - We investigated the effects of EGF, IL-1 and their combination on closure of wounds inflicted on rabbit corneal epithelial cell cultures and on migration of these cells in microchemotaxis chambers. In vitro corneal epithelial wound closure depended on the applied concentrations of EGF or IL-1. Twenty-four hours after wounding, the smallest wounds were obtained with 50 ng ml-1 of EGF and 1 ng ml-1 of IL-1, respectively. The effect on wound closure of combinations of EGF and IL-1 was additive even at concentrations that were optimal for each growth factor when applied alone. We found that EGF increases the chemotactic migration of rabbit corneal epithelial cells. Cell chemotaxis depended both on the concentration of EGF and on the number of cells applied in the assay. This response to EGF was seen at concentrations that were effective in the wound closure assay. The magnitude of the chemotactic migration response was much smaller with IL-1 than with EGF. Similarly to the observations on wound closure, the effect on cell chemotaxis of combinations of EGF and IL-1 was additive. The ability of EGF, and EGF/IL-1 combinations to modulate corneal epithelial cell chemotactic migration supports migration as a possible biological mechanism of the acceleration of corneal epithelium wound closure by these drugs. PMID- 8224018 TI - Ultrastructural analysis of arrestin distribution in mouse photoreceptors during dark/light cycle. AB - Arrestin localization was studied in BALB/c mice retinas during a 12-hr dark/light diurnal cycle and under various light/dark interruptions. Intracellular distribution of arrestin in photoreceptor cells was determined by immunocytochemistry and electron microscopy. During the light phase of the diurnal cycle, arrestin was localized mostly in the rod outer segments. During the dark phase of the cycle, arrestin was localized mostly in the inner segment, nuclei and synaptic terminals. The disc domains of the rod outer segments were labeled at a low density, but conspicuous cytoplasmic regions in the outer segment were labeled at a very high density. These cytoplasmic regions were not labeled in our illuminated retinas. Hence, intrasegmental segregation within the outer segment may be influenced by environmental lighting. During dark adaptation, increase in inner segment labeling density was observed. In previous studies, decrease in outer segment and increase in inner segment labeling density in the dark, as determined by light microscopy, was interpreted as movement of arrestin from the outer to inner segments. Our present ultrastructural analysis of arrestin distribution indicates that yet undetermined amounts of arrestin accumulate in localized regions of the outer segment in the dark. The extent of movement of arrestin to the inner segment, if it occurs, remains to be established. Localization of arrestin in phagosomes indicates that at least part of the arrestin is being degraded in the pigment epithelium. PMID- 8224019 TI - A fluorescent carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in human lens. AB - A fluorescent carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) has been identified in both human normal lenses and cataractous lenses. The total amounts of PhIP in normal lenses were 50.7 +/- 36.0 pg (n = 7) while those of PhIP in senile and diabetic cataractous lenses were 143 +/- 29.2 pg (n = 10) and 148 +/- 55.0 pg (n = 10), respectively. Although it was not possible to strictly compare the contents of PhIP among normal, senile and diabetic lenses, due to the limited numbers of normal lenses available, the mean total amounts of PhIP in normal lenses were the lowest among normal, senile and cataractous lenses when allowing for differences in age. Furthermore, there was a significant correlation between total amounts of PhIP in human lenses, and age of the lens, suggesting that the content of PhIP in human lens is increased with age. In all types of lenses, the concentrations of PhIP in the insoluble-protein fraction (pg mg-1 protein) were significantly higher than those in the water-soluble protein fraction. In addition, PhIP was formed when bovine lens was incubated at 37 degrees C under the presence of creatinine and glucose for various time periods, and the yield of PhIP during incubation was found to be time-dependent. The results from our study suggest that PhIP may be one of the age-related fluorogens in the human lens. PMID- 8224020 TI - Beta-adrenergic mechanisms affect cell division and differentiation in cultured chick lens epithelial cells. AB - Primary cultures of chick lens annular pad (CLAP) cells were exposed to beta adrenergic drugs and lipid soluble cAMP analogues. In dose-dependent manners, these agents inhibited re-entry into the cell cycle when these normally post mitotic cells were cultured in the presence of serum. The inhibition of cell division was evidenced by a lack of DNA accumulation and greatly reduced tritiated thymidine incorporation into DNA. Treated cells were also largely inhibited from spreading onto the substrate. Instead, the epithelial monolayers originally placed into culture formed nearly spherical aggregates. Treated cultures continued to synthesize crystallins and exhibited increased levels of a 49-kDa cytoskeletal protein which accumulates in fiber cells. These results indicate that beta-adrenergic mechanisms may be involved in regulating cell division and terminal differentiation of lens fiber cells. PMID- 8224021 TI - Photoreceptor degeneration in vitamin A deprivation and retinitis pigmentosa: the equivalent light hypothesis. AB - Long-term exposure of the retina to constant illumination is known to produce irreversible degeneration of photoreceptors. We propose that similar mechanisms may be involved in photoreceptor degeneration produced by vitamin A deprivation and some forms of retinitis pigmentosa (RP). Evidence is reviewed suggesting that the free opsin present during vitamin A deprivation or the mutated opsin present in some forms of RP excite the visual transduction cascade. This would produce a constant 'equivalent light' that triggers photoreceptor degeneration. Continuous real or equivalent light may produce outer segment degeneration by interfering with circadian processes, such as protein synthesis and disc shedding and lead to the loss of photoreceptors including those not expressing the mutant gene. PMID- 8224022 TI - Hydroxyl radical generation in the cat retina during reperfusion following ischemia. AB - There is increasing evidence that oxygen-derived free radicals are generated during the early phase of reperfusion, and account for part of the damage caused by transient ischemia in various tissues. To study this in the retina, cats were injected intravenously with sodium salicylate (100 mg kg-1), which reacts as a hydroxyl radical trap to form 2,3- and 2,5-dihydroxybenzoic acids (DHBA). Thirty minutes following injection, the retina of one eye of each animal was subjected to ischemia by intraocular pressure elevation via cannulation of the anterior chamber, while the fellow eye served as a sham-operated control. Ischemia was induced for 60 min (six eyes) and 90 min (eight eyes) followed by 5 min of reperfusion. In six other eyes, ischemia was induced for 90 min without reperfusion. After enucleation, the retinas were immediately removed, placed in ice-cold buffer and the retinal levels of 2,3- and 2,5-DHBA were quantitated by high pressure liquid chromatography, coupled with electrochemical detection. Results were normalized and expressed as ng DHBA microgram-1 salicylate mg-1 retinal protein. After 60 min of ischemia followed by reperfusion the normalized levels of 2,3- and 2-5-DHBA were no different in the experimental and control retinas. However, the levels of both 2,3- and 2,5-DHBA were significantly higher in the retinas subjected to 90 min ischemia followed by reperfusion than in the control tissues (P = 0.012 and P = 0.036, n = 8 respectively). Following 90 min ischemia without reperfusion, the normalized dihydroxybenzoate levels in the retinas were no higher than in their controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224023 TI - Red blood cell fatty acid levels in patients with autosomal dominant retinitis pigmentosa. AB - Fatty acid profiles were determined in red blood cell (RBC), plasma lipids and lipid fractions from 50 patients with autosomal dominant retinitis pigmentosa (ADRP) and 20 normal-sighted controls. In plasma of affected patients, significant increases in omega 6 long-chain derivatives compared to controls resulted in elevated fatty acid unsaturation whereas, in RBCs, significant decreases in the levels of omega 6 and omega 3 long-chain polyunsaturates and a reduced unsaturation index were demonstrated (P < 0.01). The disproportionate levels of long-chain fatty acids in plasma vs. RBCs were consistent with abnormal function of acyl group transfer from plasma lipids to target tissues such as RBCs or, possibly, retina. Metabolic lipid anomalies were evaluated in two sub-groups of patients with ADRP; ten with the highest and ten with the lowest RBC lipid levels of the omega 3 fatty acid, docosahexaenoic acid (DHA). Fatty acid product/precursor relationships in the sub-groups revealed reduced chain elongation and omega 3 fatty acid desaturation associated with reduced ratios of rod-to-cone electroretinographic amplitudes. Based on these results, we suggest that the heterogenic ADRP population includes a sub-group of patients characterized by abnormal fatty acid metabolism that may effect membrane structure and visual function. PMID- 8224024 TI - Metabolism in frog retinal pigment epithelium of docosahexaenoic and arachidonic acids derived from rod outer segment membranes. AB - Frog photoreceptor cells shed about 10% of their rod outer segments (ROS) every 4th day. Packets of these membranes are phagocytized and digested by retinal pigment epithelial (RPE) cells. Large amounts of lipids must be processed daily, especially docosahexaenoic acid (22:6n-3, DHA), the major fatty acid of these membranes. To study the metabolism of ROS lipids in RPE cells, RPE-eyecups were incubated with [3H]DHA-, [3H]arachidonic acid (AA)-, or [2-3H]glycerol-labeled ROS membranes for 2 hr, followed by a chase for up to 8 hr. Lipid extracts of RPE cells and incubation media were resolved into classes and quantitated for radioactivity. In RPE cells, the relative proportion of DHA and AA label in triglycerides (TG) increased dramatically with incubation time, although the substrate ROS membranes did not contain labeled TG. Other RPE lipids showed prominent reductions or relative little change. The percentage of radioactivity in free fatty acids (FFA) was low (< 3%) in RPE cells. In the chase media, the majority (60-80%) of DHA and AA label was found in FFA, with little radioactivity in TG or phospholipids. When RPE cells were incubated with [3H]glycerol-labeled ROS membranes which contained 22% of the label in TG, the most rapid reduction in relative radioactivity appeared in TG. We conclude that DHA and AA are released from phagocytized ROS membranes and are rapidly incorporated into RPE cellular lipids, primarily TG. This lipid class is very active metabolically, since TG derived from ROS are rapidly hydrolysed. Free DHA and AA of ROS origin are released from RPE cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224025 TI - Localization of hydroxyindole-O-methyltransferase in the retina: a re-evaluation. PMID- 8224026 TI - Acetylcholine receptors are coupled to mobilization of intracellular calcium cultured human lens cells. PMID- 8224027 TI - Festschrift in honor of Reubin Andres. PMID- 8224028 TI - Memories and thoughts of Reubin Andres. PMID- 8224029 TI - Origins of Reubin Andres as a clinical investigator. PMID- 8224030 TI - Age-related effects in T cell activation and proliferation. AB - Age-associated thymic involution manifests its effects in a variety of ways that are related to a loss of T cell function. These include the appearance of a non functional subset of T cells that increase in representation with age. Moreover there is a loss of T cell proliferative ability, a decline in the synthesis and release of interleukin-2 (IL-2), a decline in the ability of the T cell to express the IL-2 receptor, and a loss of control activity. This loss of control is demonstrated by the age-related appearance of autoantibodies and an increase in the elaboration of inflammatory cytokines such as TNF, IFN, IL-6, and TGF. A major part of the basis for the loss of T cell function is an inability of the T cell to respond to activation signals that are transmitted through the membrane binding of specific stimulatory signals. Transduction events, differentiation signals, and a loss of control mechanisms are all parts of a complicated picture of age-related immune deficiencies. PMID- 8224031 TI - The insulin receptor on activated immunocompetent cells. AB - This article reviews the corpus of work on the activated T lymphocyte insulin receptor begun in the laboratory of Dr. Reubin Andres. It reviews the pharmacokinetic properties of the lymphocyte insulin receptor and the mechanisms by which the receptor is regulated on the cell. An immunobiologic role for insulin and its receptors is described. Lastly, a novel monokine responsible for fine regulation of the T lymphocyte insulin receptor is described which might be an important immunosuppressant molecule for future studies. PMID- 8224032 TI - Beta adrenergic regulation of rat liver glycogenolysis during aging. AB - Studies from a number of laboratories demonstrate a biphasic change in beta adrenergic regulation of hepatic glycogenolysis over the life span of the male rat. The beta adrenergic response is prominent in immature animals, declines rapidly during subsequent development to a minimum by the time of young adulthood, and then reemerges during postmaturational development. Age changes in beta adrenergic-responsive adenylate cyclase activity follow a "U"-shaped curve similar to that described by changes in liver glycogenolytic responsiveness during aging. Developmental and postmaturational changes in beta adrenergic sensitive adenylate cyclase activation are related to parallel alterations in the density of beta adrenergic receptors and also to functional changes in nonreceptor components of the enzyme. The prevailing view that catecholamines stimulate hepatic glycogenolysis by an alpha adrenergic receptor-mediated, cyclic AMP-independent mechanism is based almost entirely on evidence from young adult male rats. We propose that current concepts of alpha adrenergic-responsive liver glycogenolysis underestimate a physiological role for beta adrenergic responsiveness over the majority of the life span. PMID- 8224033 TI - Adrenergic receptors, G proteins, and cell regulation: implications for aging research. AB - Aging is associated with changes, typically blunting, in response to catecholamines and activation of the sympathetic nervous system. These changes are observed in the presence of a substantial increase in circulating catecholamines. Adrenergic receptors with their linkage to guanine nucleotide binding (G) proteins and to effector molecules provide the critical components between catecholamines and tissue response. In this article, I review recent discoveries related to the expansion of size of adrenergic receptor family and G protein superfamily and the possible implication of these discoveries for research in aging. Nine subtypes of adrenergic receptors have been identified (alpha 1A, alpha 1B, alpha 1C, alpha 2A, alpha 2B, alpha 2C, beta 1, beta 2, beta 3), and other subtypes may yet be identified by molecular cloning techniques. The functional role of all of these receptors remains ill-defined. For the heterotrimeric G proteins, at least 16 alpha, 4 beta, and 4 gamma subunits have been identified. The G alpha subunits are unique among G proteins and have been divided into four principal families, termed Gs, G(i), Gq, and G12. G proteins are multifunctional and can link to multiple effectors. Although it is probable that beta-adrenergic receptors preferentially link to Gs, alpha 1-adrenergic receptors to members of the Gq family, and alpha 2-adrenergic receptors to members of the G1 family, many unanswered questions remain as to the determinants of these linkages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224034 TI - Similar effects of saponin treatment and aging on coupling of alpha 1-adrenergic receptor-G-protein. AB - The effects of the nonionic detergent saponin on alpha 1-adrenergic signal transduction were investigated using rat parotid cells and membrane preparations. Fifty microM epinephrine-stimulated 45Ca2+ efflux and inositol 1,4,5-triphosphate (Ins[1,4,5]P3) production in adult parotid cells were significantly decreased after saponin treatment. Saponin did not alter the concentration of alpha 1 adrenergic receptors labeled by [3H]prazosin, but significantly reduced the guanosine imido diphosphate (GppNHp)-induced shift from high to low affinity sites. Fifty microM epinephrine-stimulated high affinity GTPase activity was also reduced by saponin treatment. These data suggest that reduced alpha 1-adrenergic receptor-stimulated functional responsiveness following saponin treatment may be due to impaired uncoupling of receptor-G-protein complexes. PMID- 8224035 TI - Evaluation of hypoglycemic counterregulation using a modification of the Andres glucose clamp. AB - The glucose clamp, developed by Andres for the quantification of insulin action and secretion, was modified to study counterregulatory mechanisms against hypoglycemia, thereby overcoming the technical difficulties in producing a standardized, reproducible hypoglycemic stimulus. Normal subjects are exquisitely sensitive to small decrements in glucose; levels within the normal range cause suppression of endogenous insulin (approximately 4.0 mM) and activation of glucagon and epinephrine (approximately 3.5 mM) secretion. The glucose threshold for hormone release is modified by multiple factors, including age, gender, and the level of insulin per se. If glucose continues to fall toward 3.0 mM, the hormonal response is intensified and symptoms appear. The window between the appearance of symptoms triggering carbohydrate ingestion and the earliest signs of neuroglycopenia are surprisingly narrow. Subtle neuroelectrophysiological changes in cortical and brain stem function are evident at 2.9 mM. Insulin dependent diabetes mellitus (IDDM) patients must rely heavily on their ability to secrete epinephrine to overcome a defective glucagon response. Unfortunately, this defense mechanism may diminish as disease duration increases, and may become further impaired by iatrogenic hypoglycemia accompanying insulin treatment. Commonly, the glucose level triggering adrenergic responses is shifted downward during intensive insulin therapy of IDDM. This may help explain why such patients release epinephrine and experience symptoms of hypoglycemia at a lower glucose level. PMID- 8224036 TI - The effects of single and sequential insulin infusions on glucose disposal in older men. AB - The hyperinsulinemic euglycemic glucose clamp is widely used to quantitate in vivo insulin action. Modification of this technique by sequentially infusing multiple doses of insulin allows determination of insulin sensitivity and maximal responsiveness; however, the validity of this approach has not been determined in older individuals. In this study, glucose disposal rates during a sequential three-dose clamp at insulin infusion rates of 20, 100, and 500 mU/m2.min were compared to those obtained during a single-dose 100 mU/m2.min clamp in eight healthy older men. There were no differences in plasma insulin levels (256 +/- 46 vs. 261 +/- 32 microU/ml) or glucose disposal rates (11.0 +/- 3.6 vs. 10.8 +/- 3.0 mg/kgfat-free mass.min) during the 100 mU/m2.min infusion of the three-dose clamp and the one-dose clamp. In four subjects with impaired glucose tolerance, the EC50 (insulin concentration producing a half-maximal response) was higher and Mmax (maximal glucose disposal) lower than in subjects with normal glucose tolerance, suggesting impairments in both insulin sensitivity and responsiveness in older subjects with impaired glucose tolerance. Future studies using this modification of the hyperinsulinemic euglycemic glucose clamp to determine insulin sensitivity and responsiveness may lead to an improved understanding of the insulin resistance of aging and the pathogenesis of diseases such as diabetes and hypertension. PMID- 8224037 TI - The effect of age on insulin response and glucose utilization during four hyperglycemic plateaus. AB - In order to evaluate the potential role of insulin insensitivity as a cause of the glucose (G) intolerance of aging, we performed 230 hyperglycemic clamps, 85 on young (Y, 24 to 39 years), 47 on middle age (M, 40 to 59 years), and 98 on old (O, 60 to 90 years) carefully screened subjects of the Baltimore Longitudinal Study of Aging. The 2-h plasma G levels on an oral glucose tolerance test (OGTT) were < 7.8 mmol/l in Y and M and < 10 mmol/l in old; the latter group was further dichotomized at 7.8 mmol/l into a "normal" group, ON, and an impaired group, OI. Four hyperglycemic plateaus were created: 3.0, 5.4, 7.9, and 12.8 mmol/l above basal. Three measures of glucose tolerance--1) G at 2 h after glucose ingestion, 2) glucose utilization, M, at each hyperglycemic plateau, and 3) glucose decay constant, K, obtained at the conclusion of each clamp--showed the best performance in the young group (Y > M = ON > OI). Despite these differences in glucose tolerance, plasma insulin responses (I) during the clamp were not significantly different except that ON < Y at the basal + 12.8 plateau (300 +/- 42 vs. 456 +/- 48 pmol/l, p < 0.01). Insulin-dependent glucose uptake, a measure of tissue sensitivity to insulin, was decreased in the old-impaired group at every plateau except the highest. We conclude that healthy, active older subjects showed moderate intolerance to oral and IV glucose and that the mechanism of this physiological aging process is most likely decreased insulin sensitivity. PMID- 8224038 TI - Adverse effects of abdominal obesity on lipoprotein lipids in healthy older men. AB - Individuals with abdominal obesity are at increased risk for the development of coronary artery disease (CAD). This study examines whether an abdominal fat distribution, indexed by the waist to hip ratio (WHR), is associated with an atherogenic lipoprotein lipid profile independent of the degree of obesity (percent body fat) and maximal aerobic capacity (VO2max) in 127 older (60 +/- 8 years, mean +/- SD) normotensive, nonsmoking, nondiabetic men. Compared to men with WHR below the population mean (< 0.96, low WHR), men with high WHR (> 0.96) had higher triglycerides (TG) (124 +/- 47 vs. 93 +/- 48 mg/dl, p < 0.001) and lower high density lipoprotein cholesterol (HDL-C) (34 +/- 7 vs 41 +/- 9 mg/dl, p < 0.001) levels with no difference in low density cholesterol (LDL-C) levels. Plasma TG levels were positively associated with both percent body fat (r = 0.50, p < 0.0001) and WHR (r = 0.49, p < 0.0001), and negatively with VO2max (r = 0.36, p < 0.0001), whereas plasma HDL-C levels were negatively associated with percent body fat (r = -0.50, p < 0.0001) and WHR (r = -0.54, p < 0.0001), and positively with VO2max (r = 0.45, p < 0.0001). Independent relationships between WHR and both plasma TG (r = 0.30, p < 0.001) and HDL-C (r = -0.34, p < 0.0001) remained after correction for both percent body fat and VO2max. Using analysis of covariance, the differences in TG and HDL-C between groups remained after adjustment for percent body fat and VO2max. These results suggest that in older men, an abdominal distribution of body fat, independent of both percent body fat and VO2max, is associated with elevated TG and low HDL levels, thus increasing the risk for CAD. PMID- 8224039 TI - Aging and the response to inhibition and induction of theophylline metabolism. AB - The twofold to threefold higher incidence of adverse drug reactions in elderly as opposed to younger patients is due mainly to more severe disease and the requirement for more complex drug treatment regimens. The incidence of adverse drug reactions increases with the number of prescribed drugs. Because of multiple drug use by the elderly, the potential for drug interactions is greater in this patient group. Surprisingly, the effect of age on the clinical pharmacology of drug interactions has not been thoroughly investigated. Our studies have shown that cimetidine inhibits and phenytoin induces the metabolism of theophylline to a similar extent in healthy male nonsmokers and smokers. Preliminary analysis of the results of a study to investigate the inhibition of theophylline metabolism by cimetidine and ciprofloxacin administered in combination to healthy male and female nonsmokers also does not show an age difference in response. Additional careful studies are needed to evaluate further the pharmacology and clinical importance of pharmacokinetic and pharmacodynamic drug interactions in the elderly. PMID- 8224040 TI - Androgens and aging in men. AB - Androgen levels decrease with aging in men. Androgen deficiency in elderly men may lead to asthenia, decrease in muscle mass, osteoporosis, decrease in sexual activity, and, in some cases, changes in mood and cognitive function. Combination of these factors may result in impaired quality of life in the elderly male. Androgen replacement therapy may increase bone and muscle mass, enhance muscle and cardiovascular function, and improve sexual function and general well-being. These potential benefits of androgens have to be weighed against the possible adverse effects on prostate and cardiovascular diseases. Careful long-term studies will be required to assess the risk-to-reward ratios of androgen or other hormone replacement therapy before the development of treatment strategies similar to estrogen and progestagen substitution therapy for the postmenopausal female. PMID- 8224041 TI - Comprehensive geriatric assessment in the outpatient setting: population characteristics and factors influencing outcome. AB - The chronic diseases of old age reduce the "active life expectancy" of older persons and result in significant health care expenditures. Yet, among disabled persons, the degree of disability does not explain who actually accesses long term care (LTC) services. Comprehensive geriatric assessment (CGA) has been proposed as the technology of geriatric medicine which will reduce both excess disability and health care costs. Outpatient CGA programs may be the ideal setting in which to study older disabled persons who are at risk for increasing their use of LTC services. A prospective study of our CGA population suggests that understanding the factors which influence outcome will be critical to designing interventions and improving outcome for this population. CGA patients suffer from a limited number of functionally disabling problems, each of which has a broad array of etiologies. Use of LTC services seems most closely tied to social factors, including the feelings of burden experienced by caregivers. Caregiver burden is a better predictor of service use than any measure of the older person's mental or physical health or functional abilities. Reducing health care costs including nursing facility placement will likely require interventions which reduce caregiver burden. Further definition of the problems seen in the CGA population and delineation of the factors influencing outcome will identify interventions to achieve the desired health care outcomes. PMID- 8224042 TI - Surgery in the elderly: observations on the pathophysiology and treatment of cholelithiasis. AB - Biliary tract disease is particularly prevalent in older persons, and is the leading indication for acute abdominal surgery in the elderly. The complications of biliary stone disease are more insidious in the elderly, and the recognition of "symptomatic" disease is often delayed. The unreliable manifestations of biliary tract pathology result in a higher morbidity and mortality of the disease, and a large number of emergency operations. Recent studies suggest that the pathophysiology of cholelithiasis is specifically altered in the elderly. Pigmented gallstones, which are more common in the elderly, are thought to result from the action of bacterial enzymes that deconjugate bilirubin and form insoluble bile salts; bacterial contamination of the extrahepatic biliary system is more frequent in elderly patients. Gallbladder stasis may be due to age related changes in the response of gallbladder musculature to the kinetic action of cholecystokin (CCK), possibly through a decrease in CCK receptor affinity or availability. Other hormonal factors which inhibit choleresis, such as pancreatic polypeptide, are increased with aging as well. Current efforts are being directed toward closer surveillance and discovery of cholelithiasis, and improved outcomes of therapy have been aided by the development of "minimally invasive" approaches to the treatment of cholelithiasis in the elderly. PMID- 8224043 TI - Does diet provide adequate amounts of calcium, iron, magnesium, and zinc in a well-educated adult population? AB - Standard advice from dietitians, nutritionists, and physicians is that if one eats a well-balanced diet containing a variety of foods, supplements are not necessary. Little information is available, especially in those over 75, to determine whether actual diets do provide adequate amounts of these minerals. The participants of the Baltimore Longitudinal Study of Aging provide seven-day records which include vitamin and mineral supplement intakes. Median daily dietary intakes from diet in all 564 subjects and from diet plus supplements in those who use them were analyzed by age group and gender. More women than men took supplements. Median intakes of calcium from diet were below the recommended dietary allowance (RDA) for unsupplemented women and for supplemented women over 60. Approximately 25% of women under 50 and 10% of women over 50 consumed less than two thirds of the RDA for iron from diet. For both men and women, all groups had median diet intakes below the RDA for magnesium. Forty percent of men and about half of women consumed less than two thirds of the RDA. These results indicate that many people in this well-educated, presumably well-nourished population did not consume adequate amounts of calcium, iron, magnesium, and zinc from diet. More women than men are at risk. Even those taking supplements did not consume adequate levels of some minerals. PMID- 8224044 TI - The influence of aging on spontaneous shifts of attention from external stimuli to the contents of consciousness. AB - In a series of studies using laboratory procedures and retrospective reports it has been established that with increasing age adults less frequently have unbidden task-unrelated image and thought intrusions (TUITs). TUITs--also referred to as daydreams--have been linked to the "current concerns" and "unfinished business" of the individual, and old adults have been shown to express fewer current concerns than young adults. It has also been hypothesized that selective loss of neurons in old age might interfere with thought production, resulting in fewer unbidden thoughts and spontaneous shifts of attention to them. In this article we examine the extent to which intraindividual change in the frequency of TUITs over 6 to 8 years is consistent with the decrease expected from the prior cross-sectional studies. In particular, we examine the frequency of daydreams based upon retrospective self-reports using the Daydreaming Frequency scale of the Imaginal Processes Inventory. The longitudinal sample consisted of 93 women and 169 men. Significant and equivalent decreases in Daydreaming Frequency scale values occurred at all ages. Longitudinal decreases were consistent with cross-sectional age differences. Thus, spontaneous shifts of attention to the contents of consciousness were seen to decrease over a 6 to 8 year interval within individuals--a result consistent with a within-individual change in conditions leading to spontaneous shifts. PMID- 8224045 TI - Is there any relationship between aluminum and Alzheimer's disease? AB - The controversial role of aluminum in Alzheimer's disease (AD) is reviewed. While current data would suggest the lack of a causative role, alterations in the brain and other organ systems caused by AD might increase the penetration of aluminum as well as other metals into the brain and lead to their contribution to such pathological features as neurofibrillar tangles (NFTs). PMID- 8224046 TI - Autonomic behavior. AB - This article summarizes a body of work which collectively shows that autonomic responses meet the criteria for behavior. They can be modified reliably through the systematic use of antecedent (cues) and consequent (contingencies) stimuli. This means that autonomic responses, which are usually characterized as elicited reflexes, can be learned responses (viz., behaviors). This review cites a number of experimental and clinical studies in which autonomic learning has been shown to occur and to have clinical importance. Of special interest to gerontologists are the clinical studies which show that incontinent and hypertensive elderly patients can be trained to normalize their pathognomic responses. PMID- 8224047 TI - The effects of sound overexposure on the spectral response patterns of nucleus magnocellularis in the neonatal chick. AB - Spectral response plots from single cells in the chick nucleus magnocellularis were obtained following a 48 h exposure to a 0.9 kHz pure tone at 120 dB sound pressure level and after a recovery period of 12 days. Immediately after removal from the exposure, a variety of changes in the spectral response patterns of nucleus magnocellularis cells were noted. In particular, at recovery day 0, there was a significant decrease in spontaneous rate; a substantial loss of threshold sensitivity and frequency selectivity; and alterations in both the slope and dynamic range of the rate-intensity function. Interestingly, the maximum discharge rate appeared unaffected by the intense sound. Twelve days after removal from overstimulation, auditory function appeared to be operating at normal levels of efficacy as spontaneous rate, threshold sensitivity, frequency selectivity, and the dynamic range of the rate-intensity function were statistically identical to similar measures in nonexposed control birds. The loss and restoration of auditory function is correlated to structural damage and repair of the avian basilar papilla. PMID- 8224048 TI - Principal cells of the rat medial nucleus of the trapezoid body: an intracellular in vivo study of their physiology and morphology. AB - The medial nucleus of the trapezoid body (MNTB) is one of several principal nuclei in the superior olivary complex (SOC) of mammals. It is classically thought to function as a relay station between the contralateral ventral cochlear nucleus and the lateral superior olive (LSO), playing a role among those brainstem nuclei that are involved in binaural hearing. In order to characterise the physiology and morphology at the cellular level of the major neuronal component of the MNTB, the principal cells, we have analysed these neurons in rats in vivo using intracellular recordings and horseradish peroxidase-labelling. Our data demonstrate that MNTB principal cells, when being stimulated acoustically via the contralateral ear, show a phasic-tonic response with an onset latency of 3.5 ms and a suppression of their spontaneous activity following stimulus offset. These neurons have an axonal morphology whose complexity has not yet been described. All cells (n = 10) projected exclusively ipsilaterally and had terminal axonal arbors in a variety of auditory brainstem nuclei. At least two and maximally seven auditory targets were innervated by an individual cell. Each cell projected into the LSO and the superior paraolivary nucleus (SPN). Additional projections that were intrinsic to the SOC were often observed in the lateral nucleus of the trapezoid body and in periolivary regions, with only one cell projecting into the medial superior olive. Most, if not all, MNTB principal cells also had projections that were extrinsic to the SOC, as their axons ascended into the lateral lemniscus. In two neurons the ascending axon formed terminal arbors in the ventral nucleus of the lateral lemniscus, and the dorsal nucleus of the lateral lemniscus could be identified as a target of one neuron. The location of the cell bodies of the MNTB principal cells correlated with the neurons' best frequencies, thereby demonstrating a tonotopic organisation of the MNTB, with high frequencies being represented medially and low frequencies laterally. The axonal projections into the LSO and the SPN were also tonotopically organised and the alignment of the tonotopically organised and the alignment of the tonotopic axes was similar to that in the MNTB. Our results confirm previous data from other species and suggest that MNTB principal cells have a great amount of physiological and morphological similarities across mammalian species. Furthermore, the complexity of the axonal projections indicates that these neurons play a role in auditory information processing which goes far beyond their previously described classical role. PMID- 8224049 TI - Evidence for corrective effects of afferent signals from the extraocular muscles on single units in the pigeon vestibulo-oculomotor system. AB - The role of extraocular muscle (EOM) afferent feedback signals in the control of eye movement is still controversial. We recorded from 106 single units in the vestibular nuclei, oculomotor nuclei and reticular formation of 80 decerebrate, paralysed pigeons. EOM afferents were stimulated by passive eye movement (PEM) during vestibular stimulation by sinusoidal oscillation in the horizontal plane. We found that EOM afferent signals profoundly modified the vestibular responses of 91 (86%) of the single units recorded. As well as using PEM to simulate eye movements similar to saccades, we moved the eye in a manner which mimicked the slow phase of the vestibulo-ocular reflex (artificial VOR, AVOR). We have found evidence that, as well as providing signals closely related to the parameters of eye movement, PEM alters the vestibular responses of cells during AVOR in a manner which suggests that EOM afferent signals may play a corrective role in the moment-to-moment control of eye movement in the vestibulo-ocular reflex. PMID- 8224050 TI - Electromyographic evidence of two functional subdivisions in the rhesus monkey's flexor digitorum profundus. AB - The main belly of the macaque's flexor digitorum profundus (FDP) is divided by a dissectible plane into radial and ulnar regions. The present report describes three findings which suggest that the radial and ulnar regions represent separate functional subdivisions of the FDP. First, electromyographic (EMG) recordings during individuated finger movements performed by rhesus monkeys demonstrated different patterns of activation in the radial versus the ulnar region of the FDP. Second, studies of single motor units discriminated from the parent EMG activity also suggested at least two differentially activated motoneuronal pools in the radial versus ulnar region. Third, the finger movements evoked by intramuscular stimulation, delivered through the recording electrodes, indicated that contraction of the radial versus ulnar region produces different patterns of tension on the finger tendons. Together these findings suggest that the radial and ulnar regions of the FDP provide differential tension on the finger tendons to individuate finger movements. PMID- 8224051 TI - Reticulospinal actions on primary afferent depolarization of cutaneous and muscle afferents in the isolated frog neuraxis. AB - The effects of the brainstem reticular formation on the intraspinal excitability of low threshold cutaneous and muscle afferents were studied in the frog neuraxis isolated together with the right hindlimb nerves. Stimulation of low threshold fibers (less than two times threshold) in cutaneous nerves produced short latency, negative field potentials in the ipsilateral dorsal neuropil (200-400 microns depth) that reversed to positivity at deeper regions (500-700 microns). Stimulation of low threshold fibers (less than two times threshold) in muscle nerves produced, instead, negative response that acquired their maximum amplitude in the ventral neuropil (700-900 microns depth). These electrophysiological findings suggest, in agreement with observations in the cat, that low threshold cutaneous and muscle afferents end at different sites in the spinal cord. Intraspinal microstimulation applied within the dorsal neuropil produced antidromic responses in low threshold cutaneous afferents that were increased in size following stimulation of the dorsal or ventral roots, as well as of the brainstem reticular formation. This increase in excitability is interpreted as being due to primary afferent depolarization (PAD) of the intraspinal terminals of cutaneous fibers. Antidromic responses recorded in muscle nerves following intraspinal stimulation within the ventral neuropil were also increased following conditioning stimulation of adjacent dorsal or ventral roots. However, stimulation of the bulbar reticular formation produced practically no changes in the antidromic responses, but was able to inhibit the PAD of low threshold muscle afferents elicited by stimulation of the dorsal or ventral roots. It is suggested that the PAD of low threshold cutaneous and muscle afferents is mediated by independent sets of interneurons. Reticulospinal fibers would have excitatory connections with the interneurons mediating the PAD of cutaneous fibers and inhibitory connections with the interneurons mediating the PAD of muscle afferents. Although our results provide no direct information on whether the reticulospinal depression of the PAD elicited in low threshold muscle afferents is due to inhibition along the pathways producing PAD of muscle spindle or of tendon organ afferents, it seems likely-by analogy with what has been seen in the cat spinal cord-that these inhibitory actions are mostly restricted to the pathways producing PAD in the terminal arborizations of muscle spindle afferents. These results emphasize the specificity of the descending control of the synaptic efficacy of low threshold cutaneous and muscle afferents which could be of importance for motor performance. PMID- 8224052 TI - Component-specific effects of physostigmine on the cat visual evoked potential. AB - Pattern visual evoked potentials (VEPs) were recorded from the pial surface of the cat primary visual cortex prior to and following the intravenous administration of physostigmine, an agent which blocks the enzyme responsible for the breakdown of synaptically released acetylcholine. The control VEP was composed of a small initial positive deflection (P1), a subsequent large negative wave (N1) and a second large positive wave (P2). Following physostigmine, the amplitude of P1-N1 was diminished whereas that of N1-P2 increased. These effects were long lasting and were blocked by prior treatment with scopolamine, a result consistent with mediation by a muscarinic cholinergic pathway. Waveform subtraction revealed that the physostigmine-sensitive component had a slow, negative polarity waveform while the physostigmine-insensitive component was also slow, but positive in polarity. The fundamental nature of these components remains to be assessed. Nevertheless, the results indicate that waveforms of different polarity combine algebraically to yield the conventional VEP. PMID- 8224053 TI - Correlation analysis of respiratory neuron activity in ventrolateral medulla of brainstem-spinal cord preparation isolated from newborn rat. AB - Cross-correlation analysis was used to study functional connections between one inspiratory (I) neuron and another, and between one pre-inspiratory (Pre-I) neuron and another, in 54 brainstem-spinal cord preparations isolated from newborn rats. Pre-I neurons usually fired in the pre- and post-inspiratory phases. Neurons were recorded extracellularly with pairs of microelectrodes placed on the same or opposite sides of the brainstem. Fourteen pairs of Pre-I neurons recorded bilaterally in the rostral ventrolateral medulla (RVL), 14 pairs of ipsilateral Pre-I neurons in the RVL, 14 pairs of bilateral I neurons in the RVL and 12 pairs of ipsilateral I neurons in the ventrolateral medulla were studied. Cross-correlation histograms (CCHs) were computed. Significantly high peak bin counts were detected in 24 of 54 pairs. Peaks on one side of the origin of the CCHs were observed for one pair of ipsilateral Pre-I neurons, four pairs of bilateral I neurons and five pairs of ipsilateral I neurons. These findings suggest mono- or oligo-synaptic excitatory connections between paired neurons or shared inputs. Only one trough suggesting an oligo-synaptic inhibitory connection was evident in a CCH obtained from the pair of bilateral I neurons. This CCH revealed the peak and the trough on opposite sides of the origin, which was consistent with reciprocal excitatory and inhibitory connections between recorded neurons. Peaks on both sides of the origin were observed for three pairs of bilateral I neurons. From auto-correlation analysis and the latencies of these peaks, two of the three CCHs were consistent with reciprocal excitatory connections between recorded neurons, whereas the other CCH suggests shared inputs. Peaks at the origin were observed for two pairs of ipsilateral Pre-I neurons, four pairs of bilateral I neurons and five pairs of ipsilateral I neurons. These results suggest shared inputs. For Pre-I neurons recorded in opposite sides, no significant bin counts were detected. Peaks on one side were detected for three pairs. Present results suggest short-term synchronisation of I neurons, and of Pre-I neurons via excitatory coupling, and the likelihood of comparatively strong interaction between I neurons, which may be important in maintaining the I burst. PMID- 8224054 TI - Candidate premotor neurones of skin reflex pathways to T1 forelimb motoneurones of the cat. AB - This study explored the locations and input-output properties of a large population of putative premotor neurones of skin reflex pathways in the cat. These neurones, interneurones excited by forelimb skin afferents and antidromically from the T1 motor nucleus (MN) and/or the lateral funiculus (LF, C8/T1 border), termed antidromic cells, were extracellularly recorded at C6-8. Selection of this site was based on data showing that cells retrogradely HRP labelled from the T1 MN were most numerous in C6-8 and the observation that transection of LF at the C8/T1 border abolished most skin-evoked postsynaptic potentials of T1 motoneurones. Antidromic cells were located in laminae IV-V, VI and VII. The latencies of antidromic excitation ranged from 0.4 to 1.8 ms, with a tendency for laminae IV-V cells to show longer latencies than laminae VI and VII cells. Latency of skin-evoked excitation ranged from 0.6 ms (IV-V cells), 0.8 ms (VI) and 1.4 ms (VII) to greater than 5 ms. The sum of the ortho- and antidromic latencies (estimated central latency) of individual cells explained the central latencies of skin-evoked postsynaptic potentials in T1 motoneurones. Skin-evoked firing responses (average of eight to ten cells) were earliest and largest in laminae IV-V antidromic cells, and latest and smallest in lamina VII cells. The antidromic cells who received inputs from muscle afferents and descending tracts. The following three results support the suggestion that the sampled antidromic cells are mostly premotor neurones. (1) Projection to the T1 MN via LF was verified in six laminae IV-VII antidromic cells, as tested with threshold mapping for antidromic excitation. (2) Three skin-excited axons of the middle LF projected to T1 MN, as revealed by intra-axonal staining (HRP). (3) PHA-L injection in laminae I-V of C8 anterogradely labelled terminals in lamina IX and LF axons at T1. It is suggested that last-order neurones of skin reflex pathways to T1 motoneurones are widely distributed in laminae IV-VII of C6-8 and consist of a variety of neurones with different locations and input patterns. PMID- 8224055 TI - Open-loop and closed-loop control of posture: a random-walk analysis of center-of pressure trajectories. AB - A new conceptual and theoretical framework for studying the human postural control system is introduced. Mathematical techniques from statistical mechanics are developed and applied to the analysis and interpretation of stabilograms. This work was based on the assumption that the act of maintaining an erect posture could be viewed, in part, as a stochastic process. Twenty-five healthy young subjects were studied under quite-standing conditions. Center-of-pressure (COP) trajectories were analyzed as one-dimensional and two-dimensional random walks. This novel approach led to the extraction of repeatable, physiologically meaningful parameters from stabilograms. It is shown that although individual stabilograms for a single subject were highly variable and random in appearance, a consistent, subject-specific pattern emerged with the generation of averaged stabilogram-diffusion plots (mean square COP displacement vs time interval). In addition, significant inter-subject differences were found in the calculated results. This suggests that the steady-state behavior of the control mechanisms involved in maintaining erect posture can be quite variable even amongst a population of age-matched, anthropometrically similar, healthy individuals. These posturographic analyses also demonstrated that COP trajectories could be modelled as fractional Brownian motion and that at least two control systems-a short-term mechanism and a long-term mechanism-were operating during quit standing. More specifically, the present results suggest that over short-term intervals open loop control schemes are utilized by the postural control system, whereas over long-term intervals closed-loop control mechanisms are called into play. This work strongly supports the position that much can be learned about the functional organization of the postural control system by studying the steady-state behavior of the human body during periods of undisturbed stance. PMID- 8224056 TI - Non-uniform temporal scaling of hand and finger kinematics during typing. AB - We examined the manner in which the keystroke kinematics of the hand and the fingers varied with the mean rate of typing by trained typists. We used words and phrases in which only one letter was typed with the right hand and all of the remaining letters were typed using the left hand. We varied the typing rate over a threefold range (intervals between keypresses ranging from 150 ms to 500 ms) with the aid of a metronome. The results from four subjects, and three letters (n, u, and o) were analyzed. We did not find a simple scaling that could account for variations in the velocity profiles with typing rate. For some subjects and some letters, the velocities were independent of typing rate. In other instances, the kinematics did depend on typing rate, but to a much greater extent prior to the time of keypress than afterward. Sometimes the velocity profiles of all of the fingers and of the hand changed in a similar manner as the interval between keypresses was varied. In other instances only the focal movement of the hand and the finger used to press the key depended on the interval, whereas the motions of the other fingers did not. We suggest that the consistencies in the velocity profiles which we observed may simplify the problem of arranging a temporally ordered sequence of goal-directed movements. PMID- 8224057 TI - Facilitation of somatosensory evoked potentials by exploratory finger movements. AB - Modification of somatosensory processing depending on the behavioral setting was studied. Active alternating movements of the fingers, passive tactile stimuli to the hand, and active exploration of objects were performed during recording of somatosensory evoked potentials (SEPs). SEPs were elicited by compound electrical median nerve stimulation and electrical stimulation at detection threshold of cutaneous median nerve fascicles identified by microneurography. Electrical stimulation was not time-locked to the studied condition. In comparison with SEPs at rest there was attenuation of early cortical potentials up to 25 ms post trigger in all nonresting conditions. In stimulation of the compound median nerve as well as of isolated cutaneous fascicles of a hand actively exploring an object there was an additional increased negativity, peaking at 28 ms. This facilitory effect was independent of attentional focusing and was absent during exploration using the ipsilateral, non-electrically stimulated hand. In patients with parietal lesions the facilitatory effect was diminished on the affected side. Spline interpolated brain maps at this latency based on 32-channel recordings in healthy volunteers showed a shift of local contralateral positive maximum from frontal to parietal during exploration, indicating enhancement of a tangential dipole. It is suggested that in conditions involving close sensorimotor interaction such as exploratory hand movements there is preactivation of a cortical area which is located in the central sulcus and receives cutaneous somatosensory inputs. PMID- 8224058 TI - Egocentric references and human spatial orientation in microgravity. I. Perception of complex tactile stimuli. AB - This paper is devoted to the results of the "tactile matrix" experiment performed during the second French-Soviet spaceflight (project Aragatz). The perception of the orientation of complex tactile stimuli (letters and digits) applied to different skin areas under varied conditions was studied. The task of interpretation of complex tactile stimuli was not affected by the absence of the gravitational vertical, although this task is closely associated with mechanisms for the perception of body configuration, as well as the spatial orientation of different body parts. The number of errors made under conditions of weightlessness was often even less than on Earth. The results confirm the high stability of the egocentric reference system and provide evidence that this system is based on a body scheme which cannot be easily modified by changing external conditions. PMID- 8224059 TI - Egocentric references and human spatial orientation in microgravity. II. Body centred coordinates in the task of drawing ellipses with prescribed orientation. AB - This article describes the results of the "ellipses" experiment conducted during the second French-Soviet spaceflight (project Aragatz). The realization of oriented motor tasks, on the basis of internal body representation and without visual feedback, was chosen as a paradigm for studying the determinants of spatial orientation under weightlessness. The process of drawing ellipses in the air, using arm movements with axes parallel or perpendicular to the longitudinal body axis, was studied under normal gravity and in weightlessness, and recorded using a video computer motion-analyzing system (Kinesigraph). On Earth, the experiments were performed in standing and lying positions, and in flight, in the erect position with the feet fixed to the floor. In general, performance of the task in microgravity was not disturbed. Under conditions of spaceflight, the longitudinal ellipse was inclined forward in accordance with the inclination of the whole body relative to the fixed feet. On Earth, the angle between the long axes of longitudinal and transverse ellipses deviated from 90 degrees by 20-30 degrees. The same deviation persisted under microgravity conditions. The distinctive features of ellipses traced by individual subjects were also preserved. It is concluded that an egocentric reference system ensures normal performance of sensorimotor tasks in the absence of a gravitational reference. PMID- 8224060 TI - Proprioceptive guidance and motor planning of reaching movements to unseen targets. AB - The ability to make accurate reaching movements toward proprioceptively defined target locations was studied in seven normal subjects who were trained to reach to five different targets in a horizontal plane, with no vision of hand or target. The task consisted of moving a handle from a fixed origin to each target location, fast and accurately. Target locations were learned in training sessions that utilized acoustic cuing. Most movements were rapid, with a bell-shaped velocity profile. The error in target reproduction, which constituted the difference between the position consciously identified as the correct target location and the real target location, was calculated in each trial. This was compared with the error in preprogrammed reaching, which constituted the difference between the point in space where the initial fast movement toward the target ended and the target location. The absence of significant differences between these two error types indicated that the transformation from an internal representation of target location into a motor program for reaching to it did not introduce an additional reaching error. Learning of target locations was done only with the right hand, yet, reaching of both hands was tested. This allowed a comparison between the subjects' ability to utilize a transformed spatial code (reaching with the untrained hand) and their ability to use a direct sensory motor code (reaching with the trained hand). While transformation of the spatial code was found to reduce it's accuracy, utilization of this code in motor programming again did not appear to introduce an additional error. PMID- 8224061 TI - The suppression of cervico-ocular response by the haptokinetic information about the contact with a rigid, immobile object. AB - Horizontal eye movements were recorded in eight healthy subjects during super slow trunk rotation with respect to the space-stationary head. In some trials, subjects simultaneously indicated their perception of self-motion by means of a joystick. Over the frequency range employed (0.007-0.05 cycles per second, +/- 20 degrees), all subjects perceived the relative motion of head and trunk as a head rotation with respect to the stationary trunk. Eye movements were observed which were in phase with imaginary head rotation; their amplitude exceeded the amplitude of actual body rotation. The grasping of a rigid ground-based handle (1) produced a sensation of trunk rotation in space, (2) suppressed the sensation of imaginary head rotation in space and (3) gave rise to a significant decrease in amplitude of eye movements. The grasping of a stiff rod with non-zero compliance did not produce these effects. It is concluded that eye movements in response to body rotation with respect to the fixed head are not purely reflex reactions, but are influenced by the internal representation of body motion. PMID- 8224062 TI - The relationship between estimates of Ia-EPSP amplitude and conduction velocity in human soleus motoneurons. AB - There are several parameters associated with motoneuron size, among which are the conduction velocity of the axon as well as the size of the excitatory postsynaptic potential (EPSP) induced by stimulation of Ia afferents in the corresponding muscle nerve. In particular, it has been established in animal experiments that small motoneurons with a low conduction velocity exhibit large Ia EPSPs, whereas large motoneurons with a high conduction velocity show small Ia EPSPs. Thus small motoneurons are recruited earlier than large ones. In this study, we investigated whether such a relationship between motoaxon conduction velocity and size of the Ia EPSPs could also be found for human soleus motoneurons. In total, 36 motor units from six healthy volunteers were activated by a slight voluntary contraction and exposed to 200 stimuli of the tibial nerve in the popliteal fossa. Stimuli were delivered using a special stimulus protocol ensuring a constant pre-stimulus spike density along with a constant rate of discharge of the investigated unit. From the stimulus-correlated spike train data a measure of Ia-EPSP amplitude was obtained, along with the single-unit H-reflex latency. Additionally, for each unit, the so-called surface macro EMG was recorded, which measures the complete electrical activity attributable to the unit investigated. From the macro EMB, the intramuscular delay from arrival of each action potential at the soleus muscle and the detection of the muscle-fiber action potential picked up by the recording needle electrode were measured. All single-unit H-reflex latencies were corrected for the corresponding intramuscular delays. From the corrected latencies, single-unit conduction velocities were obtained.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224063 TI - Intracellular pH regulation in cultured rat astrocytes in CO2/HCO3(-)-containing media. AB - We studied the regulation of intracellular pH (pHi) and the mechanisms of pHi regulation in cultured rat astrocytes using microspectrofluorometry and the pH sensitive fluorophore 2',7'-bis(carboxyethyl-)-5,6-carboxyfluorescein. Control pHi was 7.00 +/- 0.02 in HCO3(-)-containing solutions at an extracellular pH of 7.35. Addition of 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) or amiloride decreased pHi, as did removal of extracellular Na+, while removal of extracellular Cl- was followed by an increase in pHi. Following exposure to an acid transient induced by increasing the CO2 content from 5 to 15%, pHi rapidly returned to base line, with an average initial rate of recovery of 0.10 pH units min-1 (corresponding to a mean acid extrusion rate of 6.3 +/- 0.36 mmolo l-1 min 1). Regulation of pHi was impaired when either amiloride or DIDS was added or Cl- was removed. This inhibition was enhanced when both DIDS and amiloride were present, and pHi regulation was completely blocked in the absence of extracellular Na+. The rapid regulation of pHi normally seen following a transient alkalinisation was not inhibited by amiloride or removal of Na+, but was partially inhibited by DIDS and by the absence of extracellular Cl-. The results are compatible with the presence of at least three different pHi regulating mechanisms: a Na+/H+ antiporter, a Na(+)-dependent HCO3-/Cl- exchanger (both regulating pHi during a transient acidification), and a passive Cl-/HCO3- exchanger (regulating pHi during transient alkalinisation). The results fail to provide firm evidence of the presence of an electrogenic Na+/HCO3- symporter. PMID- 8224064 TI - Pharmacologic characterization of opioid peptide release from chromaffin cell transplants using a brain slice superfusion method. AB - A simple chamber and an inexpensive superfusion system for studying mammalian brain slices containing neural transplants is described. With this method, rat brain slices containing bovine chromaffin cell transplants can be maintained for several hours, allowing for the determination of neurochemical characteristics and pharmacologic responsiveness of the grafted cells. Using this technique, basal and nicotine-stimulated release of metenkephalin from rat periaqueductal gray slices containing bovine chromaffin cell transplants were measured. Results showed that met-enkephalin release can be increased by nicotinic stimulation in slices containing chromaffin cell, but not control implants, for at least 8 weeks postimplantation. Furthermore, this response was dose-related. These results are in good agreement with previous behavioral studies and provide corroborative evidence for the mechanism of pain reduction by the release of opioid peptides from chromaffin cell transplants in the periaqueductal gray. This study demonstrates that neurochemical and pharmacologic analyses of neural transplants using a superfused brain slice method can be a complementary approach in determining the underlying mechanisms of neural transplants in the central nervous system. PMID- 8224065 TI - Metabolic and neuroanatomical correlates of barrel-rolling and oculoclonic convulsions induced by intraventricular endothelin-1: a novel peptidergic signaling mechanism in visuovestibular and oculomotor regulation? AB - The neuroactive peptide endothelin-1 has receptors distributed abundantly among subdivisions and nuclei of the visuovestibular and oculomotor systems. In previous work, we and others described the convulsive manifestations resulting from central injection of this neuropeptide, including nystagmus, oculoclonus, exophthalmos, tonic hindlimb extension, and a generalized repetitive motor disturbance called barrel-rolling. We applied the quantitative, autoradiographic [14C]deoxyglucose method to examine the hypothesis that visuovestibular and oculomotor structures would become metabolically stimulated when endothelin was introduced into the brain via the ventricular system in conscious rats. Since previous work had demonstrated that hypermetabolic responses to endothelin in other neural systems were inhibited by an antagonist of neuronal calcium L-type channels, nimodipine, we further tested whether the increased function of vestibulooculomotor nuclei whose metabolic activity was sensitive to endothelin could be altered following nimodipine pretreatment via the ventricle. A single unilateral injection of endothelin (9 pmol in 3 microliters saline) into a lateral ventricle provoked significantly increased rates of glucose metabolism in 22 of 39 individual anatomical structures of the visuovestibular and oculomotor systems. Among those affected were the superficial stratum of the caudal superior colliculus (+25%), the optic tract bilaterally (+35 to 43%), the oculomotor cranial nerve nuclei (III, IV, VI; range of +21 to 47%), and the medial terminal nucleus of the accessory optic tract which harbors dense fields of endothelin binding sites (bilateral increase of +70 to 96%). Several other nuclei involved in the proprioceptive and visuovestibular disturbance caused by endothelin displayed increased metabolic activity, including the cuneate, gracile, sensory trigeminal, and prepositus hypoglossal nuclei, the vestibular subnuclear system, and the cerebellar flocculus. Identification of hypermetabolic responsivity to endothelin in these structures provides further information on the anatomical substrates mediating the behavioral phenomenology of endothelin-induced motor convulsions which involve the paroxysmal participation of the extraocular muscles and motor control systems producing barrel-rolling convulsions. Nimodipine pretreatment inhibited both the convulsive activity and the cerebral hypermetabolic responses to intraventricular endothelin. The results indicate that the neural systems sensitive to intraventricular endothelin become functionally active via a calcium-mediated process that may involve the neuropeptide as an intrinsic signaling molecule. PMID- 8224066 TI - Modulatory effects of acetylcholine, serotonin and noradrenaline on the activity of cat perigeniculate neurons. AB - We studied the modulatory actions of microiontophoretically applied acetylcholine (ACH), serotonin (5-HT, 5-hydroxytryptamine) and noradrenaline (NA), and those of the adrenoceptor agonists phenylephrine (PHE, alpha 1), clonidine (CLO, alpha 2) and isoprenaline (ISO, beta) on spontaneous and visually induced activities in cat perigeniculate (PGN) and thalamic reticular (NRT) neurons (only spontaneous) during extracellular recordings performed in vivo. ACH and 5-HT were found to affect the ongoing (spontaneous) and visually evoked activity of PGN cells and also the spontaneous activity of NRT cells in an opposite fashion. ACH inhibited tonic firing and often induced burst activity. By contrast, 5-HT exerted an excitatory influence, which caused a long-lasting, very regular, high-frequency activity between about 35 and 120 Hz. Spontaneous as well as 5-HT-induced firing was found to prefer three distinct frequency ranges: 35-42 Hz, 60-67 Hz and 80 120 Hz. Opposite actions of ACH and 5-HT were also evident when applied simultaneously. ACH dampened the high-frequency activity elicited with 5-HT, and 5-HT could replace the burst activity induced with ACH application by a regular tonic activity. The absolute strength of visual responses (in spikes per second) was only slightly enhanced or reduced by ACH and 5-HT, respectively, but due to the strong effects on background activity, ACH clearly elevated the signal-to noise ratio and 5-HT reduced it. Despite its excitatory action, 5-HT did not facilitate visual responses. Spontaneous changes in ongoing activity were found to affect the visual response amplitude in the same way. Noradrenaline, the alpha 1-agonist PHE and the beta-agonist ISO exerted a weak depressant action on high frequency maintained activity, but during low-frequency single spike activity and/or burst activity a facilitatory effect was evident, which prevented the generation of burst discharges and slightly increased single spike firing. Visually evoked activity was little affected, but signal-to-noise ratio changed with changes in ongoing activity. The alpha 2-agonist CLO clearly attenuated both spontaneous activity and visual responses. We suggest that, in addition to direct effects of ACH and 5-HT on geniculate relay cells, the balance between the opposite actions of ACH and 5-HT on PGN cells determines the mode of operation in the recurrent inhibitory circuit: either a global, tonic inhibition of relay cells during a dominating 5-HT influence or a less tonic but phasic inhibition during increased activity in the cholinergic system. PMID- 8224067 TI - Visual input affects the response to roll in reticulospinal neurons of the lamprey. AB - A body orientation with the dorsal side up is usually maintained by lampreys during locomotion. Of crucial importance for this is the vestibular-driven control system. A visual input can affect the body orientation: illumination of one eye during swimming evokes roll tilt towards the source of light. The aim of the present study was to investigate the interaction of visual and vestibular inputs in reticulospinal (RS) neurons of the brainstem. The RS system is the main descending system transmitting information from the brainstem to the spinal cord. The response of neurons in the middle rhombencephalic reticular nucleus to a unilateral non-patterned optic input was investigated, as well as the influence of this input on the response of RS neurons to vestibular stimulation (roll tilt). Experiments were carried out on a brainstem preparation with intact labyrinths and, in some cases, intact eyes. Illumination of one eye or electrical stimulation of the optic nerve (10 Hz) resulted in an activation of RS neurons preferentially on the ipsilateral side of the brainstem. The same result was obtained after ablation of the optic tectum, demonstrating that there are asymmetrical visual projections to the lower brainstem which do not involve the tectum. Stimulation of the optic nerve strongly affected the vestibular response in RS neurons. As a rule RS neurons are silent at the normal (dorsal-side-up) orientation of the brainstem and become active with contralateral roll tilt. During continuous optic nerve stimulation, however, the RS neurons on the side of stimulation fire during normal orientation of the brainstem, and the response to contralateral roll tilt increases considerably in many neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224068 TI - Representation of averaging saccades in the superior colliculus of the monkey. AB - We tested the hypothesis that averaging saccades occur when two different saccades are prepared and executed simultaneously. The activity of saccade related burst neurons (SRBNs) in the primate superior colliculus was recorded while monkeys made both non-averaging saccades to single targets and averaging saccades which directed the gaze between two simultaneously presented visual targets. For movements of comparable direction and amplitude, the activity measured during averaging and non-averaging saccades was statistically indistinguishable. These results are not consistent with the hypothesis that averaging saccades result from the simultaneous execution of two different saccades at the level of the collicular SRBNs. Instead, these findings indicate that averaging saccades are represented as single intermediate movements within the topographically organized map of these collicular cells. PMID- 8224069 TI - Reduced light sensitivity of the circadian clock in a hypopigmented mouse mutant. AB - Pink-eyed dilution (p/p) is a recessive mutation in mice which results in reduced pigmentation of the retinal pigment epithelium, as well as alterations in visual pathways and function. We investigated whether this mutation also affects light information reaching the circadian clock. Entrainment to a 12 h light 12 h dark cycle and the free-running period in constant darkness were not affected by this mutation. Phase shifts in response to 1 h light pulses consisting of bright white light at either circadian time 16 or 24 also did not differ between mutant and wild-type C57BL/6J mice. However, when 5 min, 502 nm light pulses of 1.2 x 10(-1) microW/cm2 or 4 x 10(-2) microW/cm2 were given at circadian time 16, the mutant mice responded with significantly smaller phase shifts than the wild-type mice. When animals were transferred to constant light, the free-running period of wild type mice was longer than that of mutant mice, a finding which is consistent with a sensitivity difference between mutant and wild-type mice. Horseradish peroxidase tracing of retinal innervation of the hypothalamic suprachiasmatic nuclei (SCN)--the location of a circadian pacemaker--revealed a reduced innervation of the SCN in mutant mice compared with wild-type mice. The total volume of the SCN, as determined by neutral red stain, was also reduced in mutant mice, although not to as great an extent as the retinal innervation. Taken together, these results indicate that while basic characteristics of circadian clock function are not altered by the pink-eyed dilution mutation, the sensitivity of the clock to light is reduced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224070 TI - The effect of acetylcholine on the visual response of lagged cells in the cat dorsal lateral geniculate nucleus. AB - We examined the influence of acetylcholine (ACh) on the visual response properties of lagged cells in the dorsal lateral geniculate nucleus of anaesthetised cats. By means of electrophysiological techniques, the response of single cells was recorded before, during and after ionophoretic application of ACh. ACh evoked a clear enhancement of the visual response. The initial suppression that a visual stimulus evokes in lagged cells was resistant to the effects of ACh. The characteristic anomalous response component of lagged cells was also present during application of ACh. The difference in latency to half rise and to half-fall of the visual response that is found between lagged and non lagged cells was maintained during application of ACh. Taken together, the results support previous evidence from experiments with brain stem stimulation that the fundamental visual response characteristics of lagged cells are state independent. PMID- 8224071 TI - Alterations in striatal glial fibrillary acidic protein expression in response to 6-hydroxydopamine-induced denervation. AB - Following injection of 6-hydroxydopamine (6-OHDA) into one side of the substantia nigra, immunohistochemical studies showed that the number of glial fibrillary acidic protein-positive [GFAP(+)] astrocytes in the striatum was significantly increased 1 day later and reached a maximum value, with intense immunoreactivity, 4 days after 6-OHDA injection. The number of GFAP(+) cells then gradually declined but was still 1.7 times the control value by 28 days postlesion. GFAP content, determined by immunoblot, and GFAP messenger RNA (mRNA) both reached maximal increases in the striatum 7 days after lesion: the mRNA returned to control values by 28 days, whereas GFAP content remained significantly elevated. Although the increases were all larger on the lesioned side, there were also significant changes on the contralateral side, as well as following saline injection. These results support the hypothesis that products released from damaged neurons are responsible for the induction of reactive gliosis, but cannot distinguish between effects mediated directly on the astrocytes or indirectly via other cells such as the microglia. PMID- 8224072 TI - Effects of caudate nucleus stimulation on substantia nigra cell activity in monkey. AB - The present study provides evidence that the saccadic signals in the caudate nucleus (caudate) are transmitted to the substantia nigra pars reticulata (SNr). We inserted two microelectrodes into the caudate and SNr of monkeys trained to perform saccade tasks. After identifying the functional characteristics of a SNr neuron recorded, we stimulated the caudate (single pulse, < 100 microA) to see whether its discharge rate changed. Among 138 SNr cells tested, 60 showed responses to stimulation of the caudate: inhibition only (n = 21), inhibition excitation (n = 17), excitation only (n = 9), and excitation-inhibition (n = 13). The latencies were 9.0-32.5 ms (mean 16.7 ms) for the initial inhibitory responses and 6.5-35.0 ms (mean 16.7 ms) for the initial excitatory responses. Pars compacta cells (n = 10) were unresponsive. The effect of caudate stimulation was selective in terms of (1) functional type of SNr cells, (2) location of SNr cells, and (3) stimulation site within the caudate. Functional type of SNr cells: saccadic, visual, expectation-related cells were more responsive than auditory, mouth/hand/arm movement-related, and reward-related cells. Many of the cells whose functional characteristics were unidentified responded to the caudate stimulation. The preferential effects were seen among the functional subtypes: cells related to memory-guided saccades, not visually guided saccades; cells with conditioned visual responses, not simple visual responses. Location of SNr cells: the stimulus effects were seen preferentially in cells in the central part of the SNr, not in the dorsal part. Stimulus site: stronger effects, whether inhibition or excitation, were obtained when the stimulation was applied to the head-body transitional zone where visuooculomotor cells were clustered. Behaviorally contingent correlation of spike activity was found between the caudate-nigral pair of cells. For example when a SNr cell with memory-contingent saccadic activity was inhibited by the caudate stimulation, a caudate cell at or close to the stimulation site may show memory-contingent saccadic activity with a similar movement field. PMID- 8224073 TI - Paradoxical sleep deprivatory effect of a single low dose of MPTP which did not produce dopaminergic cell loss. AB - In a previous study, we reported on a selective and long-lasting paradoxical sleep (PS) deprivation in cats following repeated administration of the Parkinson syndrome-inducing neurotoxin N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). While the characteristic motor deficits occurred only from the 2nd to 3rd day of a 5-day long administration of 5 mg/kg per day MPTP i.p., the PS deprivation started immediately after the first injection and lasted altogether for 11-13 days. The motor deficiencies induced by repeated administration of MPTP are mainly due to the selective depletion of dopaminergic neurons in the nigrostriatal system as the histological and biochemical data show. The immediate onset of PS deprivation and the PS recovery, despite the definite cell loss, suggests a mechanism independent of cell destruction. In our present study we investigated the occasional histological and the PS-deprivatory effect of a single low dose of MPTP in cats. A single injection of 2 mg/kg MPTP i.p. resulted in PS deprivation lasting for 2.5-3.5 h. The duration of other sleep stages showed no significant change and PS recovery was without rebound phenomenon, as in the case of repeated administration. Even a higher single dose of MPTP (5 mg/kg) resulted in no visibly detectable nigrostriatal cell loss. We suggest that the changes in monoamine release and/or turnover are involved in the PS deprivatory effect of MPTP. PMID- 8224074 TI - Upper cervical inspiratory neurons in the rat: an electrophysiological and morphological study. AB - Upper cervical inspiratory neurons from a distinct neuronal column located near the lateral edge of the intermediate grey matter in the rostral spinal segments. Previous studies conducted in cats have demonstrated synaptic inputs to these neurons from several respiratory-related regions of the medulla, and long descending axonal projections mainly towards the motoneurons supplying the intercostal muscles. The aim of this study was to examine the electrophysiological and morphological properties of this propriospinal system in the rat. Extracellular recordings were made from 127 cervical inspiratory units, mainly in the C1 and C2 segments. Eighty-two percent could be antidromically activated from the C7/C8 border. No evidence of monosynaptic connection was obtained by cross-correlating the activity of some of these units with the discharge of the phrenic nerve. Intracellular recordings were made from seven neurons, three of which were labelled with biotinamide (neurobiotin). Long "survival times" after intracellular injections (up to 23 h) resulted in staining of axons for long distances, at least to the C5 segment. Each of the three labelled axons issued only one short collateral which arborized in the region of the phrenic nucleus. These results demonstrate that upper cervical inspiratory neurons in the rat have features similar to those previously described in the cat, including only a limited projection to the phrenic nucleus. In addition, this study provides the first morphological identification of these neurons. PMID- 8224075 TI - Torques generated at the human elbow joint in response to constant position errors imposed during voluntary movements. AB - The stiffness of the human elbow joint was investigated during targeted, 1.0-rad voluntary flexion movements at speeds ranging from slow (1.5 rad/s) to very fast (6.0 rad/s). A torque motor produced controlled step position errors in the execution of the movements. The steps began at the onset of movement, rose to an amplitude of 0.15 rad in 100 ms, and had a duration equal to movement duration. The net joint torque (muscle torque) resisting the step perturbation was computed from the applied torque, the joint acceleration, and the limb inertia. Subjects resisted the imposed step changes with approximately step changes in the net muscle torque. The mean resistance torque divided by the step amplitude was computed and is referred to as the stiffness. The stiffness increased with the voluntary movement speed, over the range of speeds (1.5-6 rad/s). The stiffness increased linearly with the magnitude of the net muscle torque on the unperturbed trials (referred to as "background torque"). The stiffness changed by only 20% when the step amplitude ranged from 0.05 to 0.15 rad. The mechanical resonant frequency (fr), estimated from the average stiffness estimates, ranged from 0.8 to 3.0 Hz. The resonant frequency approximately equaled the principal frequency component of the movement fm. On average: fr = 0.96 fm +0.46. During the fixed, 100-ms rise time of the step, the resistance was not linearly related to the background torque. At slower speeds the resistance was relatively greater during this rise time. However, when the imposed step perturbation was modified so that its rise time occurred in a time proportional to the movement duration (rather than in the fixed, 100-ms, time), the muscle torque resisting the motor during this rise time was proportional to the background torque. When these modified step responses were plotted on a time scale normalized to the movement duration, they all had approximately the same shape. Apparently the muscle viscosity scaled with the stiffness so as to maintain the constant response shape (constant damping ratio). The observed "tuning" of the mechanical properties to the movement speed is suggested to be important in the robust generation of smooth stereotyped voluntary movements. PMID- 8224076 TI - Electromyographic responses to constant position errors imposed during voluntary elbow joint movement in human. AB - The role of reflexes in the control of stiffness during human elbow joint movement was investigated for a wide range of movement speeds (1.5-6 rad/s). The electromyographic (EMG) responses of the elbow joint muscles to step position errors (step amplitude 0.15 rad; rise time 100 ms) imposed at the onset of targeted flexion movements (1.0 rad amplitude) were recorded. For all speeds of movement, the step position disturbance produced large modulations of the usual triphasic EMG activity, both excitatory and inhibitory, with an onset latency of 25 ms. In the muscles stretched by the perturbation, the early EMG response (25 60 ms latency) magnitude was greater than 50% of the activity during the unperturbed movements (background activity). In all muscles the EMG responses integrated over the entire movement were greater than 25% of the background activity. The responses were relatively greater for slower movements. Perturbations assisting the movement caused a short-latency (25-60 ms) reflex response (in the antagonist muscle) that increased with movement speed and was constant as a percentage of the background EMG activity. In contrast, perturbations resisting the movement caused a reflex response (in the agonist muscle) that was of the same absolute magnitude at all movement speeds, and thus decreased with movement speed as a percentage of the background EMG activity. There was a directional asymmetry in the reflex response, which produced an asymmetry in the mechanical response during slow movements. When the step perturbation occurred in a direction assisting the flexion movement, the antagonist muscle activity increased, but the main component of this response was delayed until the normal time of onset of the antagonist burst. When the step perturbation resisted the movement the agonist muscles responded briskly at short latency (25 ms). A reflex reversal occurred in two of six subjects. A fixed reflex response occurred in the antagonist muscle, regardless of the perturbation direction. For the extension direction perturbations (resisting movement), this response represented a reflex reversal (50 ms onset latency) and it caused the torque resisting the imposed step (stiffness) to drop markedly (below zero for one subject). Reflex responses were larger when the subject was prevented from reaching the target. That is, when the perturbation remained on until after the normal time of reaching the target, the EMG activity increased, with a parallel increase in stiffness. Similarly, when the perturbations prevented the subject from reaching the target during a 1-rad voluntary cyclic movement, the EMG and stiffness increased markedly.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8224077 TI - The effect of aging on visual-vestibuloocular responses. AB - To determine age-related changes in oculomotor function we measured vestibuloocular (VOR), optokinetic (OKN), and visual-vestibular responses in a large group of normal subjects over the age of 75 years and compared the results with those from a group of young adults. Compared with the young subjects, older subjects had: (1) an amplitude-dependent decrease in VOR gain, (2) a shorter dominant VOR time constant, (3) a lower OKN slow-phase velocity saturation, and (4) a decreased ability to enhance and suppress the VOR with vision. These functional changes in older subjects are presumably secondary to well-documented, age-related changes in the sensory and neural elements of the visual vestibulooculomotor pathways. They may be a contributory factor to the common complaints of dizziness and dysequilibrium in people over the age of 75 years. PMID- 8224078 TI - Comparison of fluctuations of motor unit recruitment and de-recruitment thresholds in man. AB - Recruitment and de-recruitment thresholds of motor units in the wrist extensor muscles can undergo important random fluctuations, even when they are measured during stereotyped contractions and relaxations. These fluctuations were statistically quantified and compared. The statistical analysis indicated that recruitment and de-recruitment thresholds display the same kind of fluctuations, and that the successive measurements are randomly distributed following a quasi normal law. We suggest that the notion of force threshold for motor unit recruitment and de-recruitment might be oversimplified and that a motor unit seems to have a range of force in which it can be recruited or de-recruited. Comparison of the mean values of recruitment and de-recruitment thresholds of the motor units in the extensor carpi radialis muscles showed that de-recruitment thresholds were significantly lower than recruitment thresholds. This difference in the thresholds, together with the difference in the motor unit discharge frequency during a contraction and a relaxation, suggests a differential control of the motoneurone activity during contractions and relaxations. PMID- 8224079 TI - Friction at the digit-object interface scales the sensorimotor transformation for grip responses to pulling loads. AB - When restraining a mechanically "active" object (one that exerts unpredictable changes in loading forces) with a precision grip of the digits, we maintain a stable grasp by modulating our grip force using somatosensory information related to the loading forces. The response to ramp load increases consists of an initial fast rise in grip force ("catch-up") followed by a secondary response that steadily increases the grip force in parallel with the load force ("tracking"). The sizes of these response components scale in proportion to the loading rate. However, maintaining a stable grasp without employing an exceedingly large grip force may require further scaling of this load-to-grip sensorimotor transformation based on two additional factors: (1) the friction at the digit object interface and (2) the grip force present at the start of the load increase. The present experiments sought to determine whether such scaling occurs and to characterize its control. Subjects restrained a manipulandum held between the tips of the thumb and index finger. At unpredictable times a pulling force appeared, directed away from the subject's hand. Each pull had a trapezoidal load profile beginning and ending at 0 N with 4-N/s ramps; each ramp was 1 s in duration. The texture of the gripped surfaces varied among sandpaper, suede, and rayon, which represented increasingly slippery surfaces. The grip force at the start of the load ramp (intertrial grip force), and the amplitudes of the catch up and secondary grip responses scaled in proportion to the inverse friction. We interpret these results to indicate a uniform scaling of the transformations controlling the intertrial grip force, the catch-up response, and the secondary response. Initial-state information from tactile cues available upon object contact appeared to update the frictional scaling value. This conclusion is based on observations of immediate changes in the intertrial grip force upon contact with a new surface, and because differences in force-rate profiles appeared virtually by the onset of the catch-up response. Similarly, the intertrial grip force also constituted initial-state information. The size of the catch-up and secondary grip force responses varied inversely with the size of the intertrial grip force. These scalings of the load-to-grip-force sensorimotor transformation for friction and intertrial grip force level appear to be functionally adaptive, because they contribute to a stable grasp (prevent object slips) while avoiding exceedingly large safety margins. PMID- 8224080 TI - Perception of horizontal head and trunk rotation: modification of neck input following loss of vestibular function. AB - Chronic loss of vestibular function modifies the role of neck afferents in human perception of self-motion. We characterized this change by comparing the self motion perception of patients with chronic vestibular loss (Ps) to that of normal subjects (Ns). Stimuli consisted of sinusoidal horizontal rotations (0.025-0.4 Hz) of the trunk relative to the head (neck stimulation) and/or of the head in space (vestibular stimulation). Perception of head rotation relative to the trunk, of trunk rotation in space, or of head rotation in space was assessed in terms of gain and phase (veridical perception, G = 1 and phi = 0 degree) as well as detection threshold using a pointing procedure. (1) Perception of head rotation relative to the trunk (neck proprioception). Ps' detection threshold of head-to-trunk rotation was normal (i.e. similar to that of Ns) across all frequencies tested. Also, with peak angular velocities above 5 degrees/s, the gain of their perception was approximately normal. When peak velocity was decreased below this value, however, either by lowering stimulus frequency with peak displacement kept constant (+/- 8 degrees) or by decreasing peak displacement at constant frequency (0.05 Hz), the gain increased above unity, unlike in Ns. In contrast, the phase remained normal (approximately 0 degree). (2) Perception of trunk rotation in space. Ps perceived their trunks as stationary during neck stimulation and all vestibular-neck combinations at medium to low frequencies. At 0.4 Hz, however, Ps consistently perceived the trunk rotation, conceivably due to somatosensory self-motion cues arising from high body acceleration. In contrast, Ns perceive a trunk-in-space rotation with the neck stimulation and most of the stimulus combinations across the whole frequency range tested. Ns perceived their trunks as stationary only during head rotation on the stationary trunk (presumed to reflect a mutual cancellation of neck and vestibular signals). (3) Perception of head rotation in space. In Ps, unlike Ns, this perception always resembled that of head rotation relative to the trunk. (4) When Ps were presented with a visual or somatosensory space reference (not motion cues), their perception of trunk and head rotation in space became approximately normal. (5) We suggest that there are basically two changes in the neck-induced self-motion perception associated with chronic vestibular loss. First, neck proprioception shows a non-linear gain that overemphasizes low stimulus velocities, for unknown reasons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8224081 TI - Neural mechanisms that contribute to cyclical modulation of the soleus H-reflex in walking in humans. AB - The amplitude of the Hoffmann reflex (H-reflex) of the human soleus muscle is modulated in a cyclical way during walking. This paper addresses two questions associated with the neural mechanisms that might generate this modulation: (1) Does the amplitude of the H-reflex simply rise and fall as a function of the background excitability of the soleus motoneuron pool? (2) Is the modulation of the H-reflex dependent on events associated with activation of the antagonist muscle? The amplitude of the soleus H-reflex was compared under three conditions: natural walking, walking without activating the tibialis anterior muscle, and walking with activation of the soleus muscle in the swing phase. Human subjects were able to perform these three tasks with minimal training. The results indicated that the soleus H-reflex remained very depressed in the swing phase of walking, even when a voluntary contraction of the soleus muscle was superimposed during this time. Moreover, the presence of tibialis anterior activity had a very minor effect on the amplitude of the soleus H-reflex during walking. It is concluded that modulation of the soleus H-reflex is not simply a reflection of the background excitability of the motoneuron pool, and the modulation is not dependent on activation of the antagonist muscle. Other more powerful mechanisms are acting to modulate the reflex, most likely presynaptic inhibition of the primary afferents. PMID- 8224082 TI - Hemoglobin denaturation and iron release in acidified red blood cell lysate--a possible source of iron for intraerythrocytic malaria parasites. AB - Intraerythrocytic malaria parasites ingest the cytosol of their host cell and digest it inside their acid food vacuoles. Acidified (pH 4-5.5, 37 degrees C) human red blood cell lysates were used to simulate this process, measuring the denaturation of hemoglobin (Hb) and the release of iron, in the absence or presence of exogenous protease. Spontaneous Hb denaturation and appearance of non heme iron were observed upon lysate acidification, its rate decreasing with increasing pH, and increasing in presence of protease. Although the pH- and proteolysis-dependent release of iron paralleled Hb denaturation, released iron accounted for only a few percent of degraded Hb. Superoxide dismutase, catalase, and various scavengers of oxidative radicals had no effect on either process, consistent with the involvement of Fe(IV) intermediates in iron release from heme. Histidine and imidazole inhibited iron release, probably by binding directly to heme. Ascorbate enhanced iron release considerably but marginally enhanced the denaturation of Hb, suggesting that redox cycling of lysate free iron accelerated further release from heme. These processes could account for the endogenous supply of iron to the malarial parasite. PMID- 8224083 TI - Trypanosoma cruzi: recognition of a 43-kDa muscle glycoprotein by autoantibodies present during murine infection. AB - The production of autoimmune antibodies during infection with the hemoflagellate parasite Trypanosoma cruzi has been previously reported by this and other laboratories. The present investigation indicates a potential target in normal heart tissue that may be the binding site for the autoimmune antibodies. Using Western blot analysis, sera obtained from chronically infected C57 B1/6 mice was examined for its ability to recognize normal tissue antigens from uninfected syngeneic animals. Antibodies from chronic sera recognized an antigen in normal mouse heart and skeletal muscle tissue at a molecular weight between 43 and 45 kilodaltons (kDa). This proposed target antigen was found in cytoplasmic extracts from normal heart antigen preparations. The autoantibody recognition of the normal tissue in Western blots was inhibited by periodate oxidation of the antigen sample. This suggests that the binding of the autoantibodies may be dependent upon a glycosylation present on the antigen targets. Isotype analysis indicated that the autoreactive antibodies are of the IgM and IgG2b isotypes. These results suggest that a target for autoantibodies in Chagas' disease may be a 43- to 45-kDa glycoprotein found in normal heart and skeletal muscle tissue. Exposure of the immune system to this antigen may follow the damaging effects of the parasite itself, or of the immune response toward the parasite. In either scenario the identification of a target in the ongoing autoimmunity may help to define the autoimmune process in American Trypanosomiasis. PMID- 8224084 TI - Schistosoma mansoni: immunoreactivity of human sera with the surface antigen Sm23. AB - Sm23, a surface antigen of Schistosoma mansoni, elicits an immune response in patients infected with the parasite, although to a lesser degree than Sj23, the homologue of Sm23 in Schistosoma japonicum. To characterize the host immune response to Sm23, we have expressed full-length Sm23 using the baculovirus expression system and have also expressed the N-terminal 133 amino acids and the 85 C-terminal amino acids by means of the prokaryotic vector pGEX. A total of 70 sera from patients from Sudan and Egypt were examined for alpha-Sm23 antibodies in an ELISA and in Western blot analysis using both the C-terminal polypeptide and the full-length protein. A subset of these sera was also tested for reactivity with the N-terminal polypeptide. The alpha-Sm23 antibody titers in infected patients varied widely and were not correlated with egg counts or age of the individuals. Most of the seroreactivity was directed against the C-terminal polypeptide. PMID- 8224085 TI - Giardia lamblia: post-translational processing and status of exposed cysteine residues in TSA 417, a variable surface antigen. AB - Giardia lamblia develops and thrives within the harsh and variable environment of the human small intestine. To survive this environment, Giardia has evolved a unique family of antigenically variable, extremely cysteine-rich surface proteins. We have characterized the expression of one of these antigens, TSA 417, at the gene and protein levels. TSA 417 mRNA and protein were expressed at constant levels during both logarithmic growth and encystation. On the trophozoite surface, TSA 417 exists in two forms, an 85-kDa protein and a derived 66-kDa form. Both species have identical amino-terminal sequences that match the translated sequence of TSA 417 after removal of a predicted 17 amino acid signal peptide. Since TSA 417 is highly cysteine rich (ca. 12 mole%), we asked whether it was complexed with itself or with other molecules. Although we found no interchain disulfide bonds, there was substantial intrachain bonding that helped retain the gross structure of isolated TSA 417 after partial trypsin digestion. Because the hydropathy profile of TSA 417 suggested that most of the cysteine and all 29 of its conserved CXXC motifs are on the cell surface, we investigated whether TSA 417 might contain some of the free thiols previously shown to be on the trophozoite surface (Gillin et al., Molecular and Biochemical Parasitology, 13, 1-12, 1984). However, no free thiols were detected, either exposed on native TSA 417 or in the unfolded protein. The absence of free thiols and resistance to proteolytic digestion suggest that most of the cysteine residues are in intrachain bridges, probably either disulfide bonds or cysteine-metal complexes. This internal crosslinking may help explain the resistance of Giardia to proteases and other degradative enzymes in the intestinal fluid. PMID- 8224086 TI - Trypanosoma brucei rhodesiense: the inhibition of HL-60 cell growth by the African trypanosomes in vitro. AB - Trypanosomiasis is of major public health importance in Africa where the disease affects man and livestock. In order to explore the underlying mechanisms of pathogenesis in African trypanosomiasis, we studied the inhibition of host cell (human promyelocytic HL-60 cells) growth by Trypanosoma brucei rhodesiense using an in vitro system. This inhibition was not due to changes in pH or nutritional depletion of the culture medium by the trypanosomes as inhibitory activity was still observed in cultures that had been supplemented with glucose or fresh culture medium. Our study suggests that the African trypanosomes produce a soluble factor which inhibits the growth of HL-60 cells. This growth inhibitor does not appear to kill the HL-60 cells as determined by the trypan blue dye exclusion test. The production of this factor does not require host cell contact nor does it require a host cell cofactor. The trypanosome growth inhibitor is strictly a trypanosome product. Estimation of the molecular weight of the trypanosome growth inhibitor with Amicon filters revealed that the factor is greater than 30,000 Da in size. Protease and heat treatment of the factor resulted in the depletion of inhibitory activity. These results indicate that the African trypanosomes produce a large-molecular-weight protein growth inhibitory factor which could play a role in the pathogenesis of the disease. PMID- 8224087 TI - Babesia bigemina: isolation and characterization of merozoite rhoptries. AB - Babesia bigemina apical membrane polypeptide p58, encoded by a multigene family with homologues in other Babesia spp. and sequence similarity to rhoptry proteins in other apicomplexan parasites, was identified within merozoite rhoptries using immunoelectron microscopy. To identify additional B. bigemina rhoptry proteins, rhoptries were isolated from French pressure cell-disrupted merozoites fractionated by differential centrifugation and isopycnic sucrose density gradient centrifugation. A fraction with a density of 1.16 g/ml contained club shaped, electron-dense, membrane-bound organelles. Organelles were identified as rhoptries based on the following criteria: (1) density and morphology similar to those of rhoptries isolated from other apicomplexan parasites, (ii) dimensions similar to those of B. bigemina rhoptries in intact merozoites, and (iii) reactivity with an anti-p58 monoclonal antibody, but not with a monoclonal antibody against the merozoite outer membrane. Immunization of mice with isolated rhoptries induced antibodies that reacted with B. bigemina merozoites in an apical immunofluorescence pattern and bound the rhoptries in immunoelectron microscopy. Immunoprecipitation of [35S]methionine-labeled B. bigemina merozoites with the anti-rhoptry mouse serum identified at least seven putative rhoptry polypeptides including p58 (apparent molecular masses of > 225, 190, 76, 69, 58, 54, 36 kDa). The anti-B. bigemina rhoptry serum did not immunoprecipitate any proteins from [35S]methionine-labeled Babesia bovis merozoites consistent with previous observations that members of the Babesia rhoptry gene families do not encode B-cell epitopes conserved between species. PMID- 8224088 TI - Leishmania donovani: in vitro evidence of hepatocyte damage by Kupffer cells and immigrant macrophages in a murine model. AB - Infection with Leishmania donovani leads to activation of liver macrophages. The role of different macrophage populations of liver in this infection is not clearly defined. Thus, the mechanism involved in hepatocyte damage was studied by coculturing hepatocytes with two populations of liver macrophages, the kupffer cells and immigrant macrophages. The results indicated maximum tissue damage at peak infection in both the macrophage populations cocultured with hepatocytes (P < 0.001). Kupffer cell-hepatocyte coculture treated with scavengers of reactive oxygen intermediates failed to inhibit the hepatocyte damage (P > 0.05). But with heparin and phenylmethylsulfonyl fluoride, a sharp decrease in the damage was noticed (P < 0.001). In contrast, immigrant macrophage-hepatocyte coculture showed a significant reduction in tissue damage when treated with both the scavengers of reactive oxygen intermediates and enzyme inhibitors (P < 0.001). Therefore the murine infection with L. donovani is speculated to involve two distinct subpopulations of liver macrophages with marked differences in morphology and functional capabilities. PMID- 8224089 TI - Trypanosoma cruzi: correlation between karyotype variability and isoenzyme classification. AB - Forty-three Trypanosoma cruzi isolates from Chile and Colombia and three cloned stocks from Bolivia and Brazil were studied at the karyotype level by hybridization with four different parasite gene probes to chromosomes separated by pulsed-field gel electrophoresis. The results showed that classification of parasite isolates based on isoenzyme analysis at 12 or more genetic loci correlated with the classification obtained by molecular karyotype analysis. However, less correlation was found between molecular karyotypes and the zymodemes Z1, Z2Bra, Z2Bol, and Z3 based on analysis at only two genetic loci. All the four probes used in this study allowed differentiation between different T. cruzi stocks but the SAPA and the antigen 13 probes were most informative. Isolates which were unclassified at the isoenzyme level were also studied and in most cases similar hybridization patterns were observed as obtained with one or more isoenzyme-classified isolates. The results demonstrate the potential of using molecular karyotyping as a tool for classification. A few pulsed-field gel electrophoresis hybridization experiments provide the same information as obtained by isoenzyme analysis using a dozen or more enzymes. The clonal theory of T. cruzi propagation is supported by our results since the strong correlation between isoenzyme classification and molecular karyotype is difficult to explain with a sexual mode of replication. No minichromosomes were detected in any of the T. cruzi samples studied. Neither was any strong correlation found between the clinical manifestations of Chagas' disease and the molecular karyotypes of the T. cruzi isolates. PMID- 8224090 TI - Schistosoma mansoni: analysis of an unusual infection phenotype in the intermediate host snail Biomphalaria glabrata. AB - Snails from a B. glabrata stock, selected for nonsusceptibility to the NMRI strain of S. mansoni, exhibited unusual schistosome infections, with multiple sporocysts prominent in the head-foot (foot-sporocysts). When F1 progeny from numerous crosses between well-defined susceptible (S) and resistant (R) parental snails were exposed, 60% of the infected snails also exhibited this unusual phenotype. F1 progeny from S x S snail crosses developed normal infections. Foot sporocysts usually developed much later than secondary sporocysts of normal infections and before 12 weeks postexposure the tissue reaction surrounding foot sporocysts was less intense than reactions to sporocysts in other tissues of the body. Cercariae were seen emerging directly from foot-sporocysts. Infection of S x R hybrid progeny by three other strains of S. mansoni also resulted in the production of foot-sporocysts. Development of the foot-sporocyst infection phenotype in progeny from many snail crosses suggests that this phenotype is more common than previously recognized. Over 50% of the F1 progeny from S x R parental crosses developed patent infections, a fact relevant to the possible biological control of schistosomiasis by the introduction of refractory snails into endemic areas. PMID- 8224091 TI - Haemonchus contortus: identification of proteases with diverse characteristics in adult worm excretory-secretory products. AB - Host tissue penetration and feeding by helminth parasites may be mediated by both mechanical processes and histolytic products released by the parasites. To investigate potential histolytic products released by adult Haemonchus contortus worms, proteases in excretory-secretory (ES) products were analyzed. The optimum activity of ES proteases was at pH 5.0, although activity was observed over a wide range of pH tested (pH 3.0-9.0). Four protease bands were observed on gelatin-containing polyacrylamide gels, with estimated molecular weights (M(r)) of 32, 35, 38, and 40 kDa. Proteases of 32 and 35 kDa were active at pH 5.0-8.0, while activity of the 38- and 40-kDa proteases was inhibited at pH 8.0. Based on inhibitor studies, the four proteases identified on gelatin-containing polyacrylamide gels were classified as cysteine proteases. Evidence was also obtained that indicated the presence of aspartic and metalloprotease activities in ES products, but these activities were not detected in gels. The diversity of adult H. contortus ES proteases may indicate variable functional requirements for the proteases. Further characterization of ES proteases will facilitate evaluation of their potential application in immunotherapeuti control of disease. PMID- 8224092 TI - Recent molecular approaches to the study of schistosome genetics. PMID- 8224093 TI - Forms and functions of nematode surfaces. PMID- 8224094 TI - Molecular characterization of cDNA sequences encoding glutathione S-transferases of Fasciola hepatica. PMID- 8224095 TI - Transcription factor IIIA (TFIIIA): an update. AB - Xenopus transcription factor, termed TFIIIA, is the first eukaryotic transcription factor purified to homogeneity and one of the most extensively characterized polymerase III gene factors at the levels both of the protein and its gene. It is an abundant protein in oocytes and is specifically required for the 5S RNA gene transcription. It promotes the formation of a stable transcription complex by first binding to the internal control region of the 5S RNA gene through its zinc finger motifs. It contains two structural domains and associates with 5S RNA to form 7S ribonucleoprotein particles in oocytes. Its expression is developmentally controlled at the level of transcription and translation. It participates in the assembly of active chromatin templates and, at least in part, is responsible for the differential expression of two kinds of 5S RNA genes in Xenopus. PMID- 8224096 TI - Ultrastructural changes in rat mammotropes following incubation with dopamine. AB - Cultured mammotropes incubated with dopamine for one hour exhibited changes in ultrastructure indicative of actively depressed biosynthetic and secretory activity. Peripheral relocation of rough endoplasmic reticulum appeared to create a barrier to secretory granule release by exocytosis. A decrease in the numbers of secretory granules indicated a decrease in prolactin production and enhanced lysosomal activity. PMID- 8224097 TI - Perception of olfactory and intranasal trigeminal stimuli following cutaneous electrical stimulation. AB - Based on previous research it may be hypothesized that the perception of odorants is modified by an axon reflex emanating from trigeminal afferents activated via the skin and/or the intranasal respiratory epithelium. The present experiment investigated the effects of trigeminal cutaneous stimulation on intensity estimates of intranasal chemical stimuli. While the left nostril was stimulated chemically with olfactory and trigeminal stimulants, four regions of the face were stimulated electrically. Intensity estimates of the chemical stimuli tended to increase after cutaneous electrical stimulation which may be interpreted in terms of response priming. The effect of electrical stimulation did not differ at the 4 stimulation sites. The results argue against the hypothesis that the processing of intranasal chemical stimuli is modified peripherally by cutaneous trigeminal excitation. PMID- 8224098 TI - Decreased rates of Ca(2+)-dependent heat production in slow- and fast-twitch muscles from the dystrophic (mdx) mouse. AB - Using a newly developed microcalorimetric approach to assess the rate of energy expenditure for intracellular [Ca2+] homeostasis in isolated muscles at rest, we found this was lower in mdx than in control mouse muscles, by 62% and 29% in soleus and extensor digitorum longus, respectively. Differences in total and Ca(2+)-dependent rates of specific heat production between mdx and control were enhanced during sustained, KCl-induced stimulation of energy dissipation. These results suggest that the low sarcoplasmic energy status of dystrophic muscles is not due to any excessive energy expenditure for intracellular [Ca2+] homeostasis. PMID- 8224099 TI - Impaired insulin release in aging rats: metabolic and ionic events. AB - We investigated the effect of aging on glucose uptake, glucose-induced O2 consumption, glucose-induced 45Ca movements, and calmodulin content to elucidate age-related impairment of glucose-induced insulin release in pancreatic islets of Wistar rats. Intact pancreatic islets from old (24-month-old) rats showed impaired glucose-induced insulin release; glucose uptake and O2 consumption were lower in old than in young (2-month-old) or adult (12-month-old) rats. Moreover, 45Ca uptake and calmodulin content were decreased in pancreatic islets from older rats, which explained the impairment in glucose-induced insulin release in aging. No major differences between the 3 age groups in glucose-induced 45Ca efflux in pancreatic islets were observed. PMID- 8224100 TI - Ca-efflux, from direct membrane injury by CCl4, is elicited by amphiphilic vehicles in vitro. AB - Direct membrane injury by CCl4, in situations excluding metabolic activation, was evaluated in saponin-permeabilized hepatocytes and in microsomes by measuring immediate Ca2+ efflux. A good correlation appears between the Ca2+ efflux and the level of CCl4 in the membrane and also the variations in fluidity. Mixtures of CCl4 with water-soluble vehicles were used to improve the dispersion of CCl4 in the medium. The mixtures varied in their ability to elicit the membrane effects of CCl4. The performance of ethanol and, to a lesser degree, other alcohols, suggests the existence of a water stable structural organization between CCl4 and these amphiphilic vehicles, facilitating the transfer of CCl4 to the membrane. PMID- 8224101 TI - The involvement of noradrenaline, 5-hydroxytryptamine and acetylcholine in imipramine-induced seizures in mice. AB - The influence of some noradrenergic, 5-hydroxytryptaminergic and cholinergic agents on imipramine-induced seizures were investigated in mice. DL-threo-3,4 dihydroxyphenylserine (DOPS) and pargyline significantly potentiated imipramine induced seizures. Phentolamine and prazosin significantly attenuated seizures elicited by imipramine and significantly attenuated the seizure-enhancing effect of DOPs. alpha-Methyl-p-tyrosine and reserpine significantly attenuated seizures induced by imipramine. Disulfiram significantly protected mice against imipramine induced seizures. However, DOPS significantly potentiated seizures induced by imipramine in disulfiram-pretreated animals. Clonidine effectively protected mice against imipramine-induced seizures. Idazoxan, on the other hand, significantly potentiated seizures induced by imipramine and significantly antagonised the protective effect of clonidine against the seizures. 5-HTP, PCPA, cyproheptadine, mianserin, ketanserin and trazodone did not affect imipramine-induced seizures to any significant extent. Physostigmine antagonised seizures induced by imipramine while atropine significantly potentiated the seizures, and significantly attenuated the protective effect of physostigmine against the seizures. These data suggest that enhancement and attenuation of central noradrenergic and cholinergic neurotransmissions respectively, and not 5-HT mechanisms, may underlie imipramine-induced seizures in mice. PMID- 8224102 TI - Blockade by zinc ions of K(+)-induced contraction and calcium in guinea pig Taenia coli. AB - Preincubation with 0.3 mM Zn2+ markedly inhibited both the tonic response and Ca2+ binding at low affinity sites induced by K+ (60 mM), with smaller effects on the phasic response and the high affinity Ca2+ sites, in Taenia coli. However, when the muscle was kept in Zn(2+)-containing medium following the first stimulation with the K+, the phasic response and the high affinity Ca2+ sites were more severely inhibited during the second stimulation with K+. This probably indicates that Zn2+ reduced the tonic tension response to K+ mainly by inhibiting Ca2+ influx at the cell membranes of Taenia coli. However, when Zn2+ is continuously present, Ca2+ is not supplied at the storage sites and is not available for the phasic response to a second stimulation with K+. PMID- 8224103 TI - Lipid peroxidation, low-level chemiluminescence and regulation of secretion in the mammary gland. AB - In mammary explants of lactating mice, changes in the intensity of chemiluminescence (CL) were observed after the addition to the incubation medium of hormones and mediators that are involved in the regulation of secretion: oxytocin, acetylcholine, epinephrine and norepinephrine. A 15-min period of treatment with oxytocin, epinephrine or norepinephrine changed the level of thiobarbituric acid-reactive substances (TBARS). Two mammary explants, one of which was treated with oxytocin, acetylcholine, epinephrine or norepinephrine, were found to interact even when separated by a quartz glass wall. Analysis of the level of TBARS formation in these two explants showed that the observed interactions might be connected with light emission resulting from lipid peroxidation (LP) processes. The possible role of LP and low-level CL in the regulation of mammary gland secretion is discussed. PMID- 8224104 TI - K252a: a new blocker of the cell-cycle at G1 phase in a human hepatoma cell line. AB - The administration of 200 nM K252a to HuH7 suppressed the proliferation of the cells almost completely. The uptake of [3H]thymidine was inhibited, and flow cytometry revealed only one peak at 2C on day 3 after treatment with 100 nM K252a. The expression of proto-oncogene c-myc was not reduced. Despite the blockage at G1, both the size of the cells and the amount of cell protein had increased by 4 times by day 3 after treatment with K252a, while the cells secreted albumin and alpha-fetoprotein into the medium as usual. These results show that K252a can increase the cell size of HuH7 without losing its function by blocking the cell cycle at G1 phase. PMID- 8224105 TI - Effects of receptor agonists on polyamine concentrations and spermidine/spermine N1-acetyltransferase activity in rat parotid gland. AB - The polyamine putrescine might be formed via a degradation (catalyzed by spermidine/spermine N1-acetyltransferase, SSAT) of the higher polyamines spermidine and spermine to putrescine. The involvement of different intracellular signal pathways in the regulation of putrescine formation was studied in explants and in cultured cells of rat parotid glands by using receptor agonists that activate separate second messenger systems, and measuring their effects on the concentrations of putrescine, spermidine and spermine and on the SSAT activity. The beta-adrenoceptor agonist isoprenaline, which is an activator of cAMP formation, increased the putrescine concentration and stimulated the SSAT activity. Pilocarpine, a drug that activates the muscarinic receptors and thereby enhances the phosphoinositide turnover, had no effect on either the polyamine concentrations or on the SSAT activity. Epidermal growth factor (EGF), which induces activation of a protein tyrosine kinase, had no effect on the polyamine concentrations or on the SSAT activity. The adenylate cyclase activator forskolin increased the glandular levels of putrescine. Taken together, these findings suggest that increases in putrescine concentration in cultured rat parotid gland cells are accompanied by accumulation of cAMP. PMID- 8224106 TI - New amino sugar analogues are incorporated at different rates into glycoproteins of mouse organs. AB - Different radiolabelled N-acyl-derivatives of D-glucosamine were synthesized using D-glucosamine and the respective carbonic acid anhydride. Metabolism of these sugar analogues could be shown in vitro as well as in vivo. After the intraperitoneal administration of these radiolabelled N-acyl-D-glucosamines to mice, their rate of incorporation into glycoproteins of different organs was found to increase markedly with the length of the N-acyl side chain. Highest incorporation was measured in the whole intestine using N-pentanoyl-D-glucosamine as label. PMID- 8224107 TI - Kinetic analysis of lactate dehydrogenase using integrated rate equations. AB - The reaction catalyzed by lactate dehydrogenase was analyzed under fully second order conditions using integrated rate equations. A two-step regression analysis was utilized to fit twenty-one progress curves repeated in sextuplicate to the general mechanism second-order integrated rate equation with additional terms of substrate inhibition. The fitting error was less than one percent. The resulting kinetic constants support a ternary complex mechanism; in no case were constants supporting another mechanism predicted. The inhibition constant for oxamate was also determined. PMID- 8224108 TI - Synthetic octapeptide pyroGLU-ASP-ASP-SER-ASP-GLU-GLU-ASN controls DNA transcription in vitro by RNA polymerase II. AB - The effect of the synthetic octapeptide pyroGLU-ASP-ASP-SER-ASP-GLU-GLU-ASN (phosphorylated by casein kinase II, CKII) on DNA transcription by RNA polymerase II has been studied. The peptide contains the acidic carboxy-terminus heptapeptide of the largest subunit of RNA polymerase II, which has been demonstrated to be a phosphorylation site for CKII. The aim of this work is to obtain some insights about the possible role of this domain in RNA polymerase II activity and DNA binding. Results demonstrated that the phosphorylated octapeptide causes strong inhibition of transcription of calf thymus DNA or pSVL SV40 plasmid DNA by RNA polymerase II, when used at concentrations between 0.4-4 micrograms/ml. PMID- 8224109 TI - PCR jumping in clones of 30-million-year-old DNA fragments from amber preserved termites (Mastotermes electrodominicus). AB - DNA from 30-million-year-old amber preserved termites (Mastotermes electrodominicus) was PCR amplified with nuclear ribosomal RNA small subunit primers and cloned into the TA vector (INVITROGEN). We obtained several classes of recombinant clones as a result. Authentic Mastotermes electrodominicus clones were identified. The source of other classes of clones was identified as contaminants of the ancient DNA template. Several of the clones appeared to be chimeric in structure with half of the clone identical to the termite sequence and the other half identical to contaminant sequences. The phenomenon of PCR jumping was identified as a possible source for the chimeric clones. PMID- 8224110 TI - Implication of C-5, C-6 unsaturation as a key structural factor in steroidal alkaloid-induced mammalian teratogenesis. PMID- 8224111 TI - Pigeon milk: a new source of growth factor. AB - Crude, partially purified and purified fractions of pigeon milk injected subcutaneously in newborn mice brought about precocious opening of eyelids by 2-3 days and eruption of incisors by 3-4 days. The biological activity of pigeon milk derived growth factor (PMGF) compared well with that of mouse epidermal growth factor (mEGF). PMID- 8224112 TI - A primate model of monotypism in atherosclerotic lesions. AB - We have characterized the expression of allelic variants of X-linked glucose 6 phosphate dehydrogenase (G6PD) in aorta from homozygous, hemizygous, and heterozygous baboons (Papio hamadryas). Fibrous plaques from heterozygous baboons fed a high cholesterol, saturated fat diet contained distributions of G6PD allelic variants that differed from those of normal arterial wall and fatty streaks. The skewed allelic expression patterns in fibrous plaques of heterozygotes reflect decreased cellular heterogeneity in advanced vascular lesions. The tendency toward cellular monotypism in fibrous plaques is similar to that present in advanced human atherosclerotic lesions. Our results suggest that G6PD heterozygous baboons are a unique primate model for investigating the cellular origin of proliferating smooth muscle cells in atherosclerotic plaques. PMID- 8224113 TI - Dietary fish oil reduces microthrombi over atherosclerotic lesions in hyperlipidemic swine even in the absence of plasma cholesterol reduction. AB - We have investigated in swine the effect of fish oil additives to a butter cholesterol hyperlipidemic diet (BT) on atherogenesis and thrombogenesis when average plasma cholesterol levels were kept similar in fish oil-treated and untreated BT groups. The studies included evaluation of lesion sizes and cell numbers, counts of adherent monocytes over lesions, and counts of platelet clumps (microthrombi) over lesions either attached directly to endothelium or to adherent monocytes. Anatomic sites studied for lesion development were the left anterior descending coronary artery (LAD), the distal 1/5 of the abdominal aorta, and a proximal portion of the thoracic aorta. Counts of attached monocytes and platelet clumps were made by scanning electron microscopy only for the LAD and expressed per mm2 of surface. The most striking new result was in regard to the platelet clumps. These were reduced by the fish oil from 996 +/- 295/mm2 in the untreated BT group to 313 +/- 59 and 364 +/- 105 in BT+cod liver oil and BT+menhaden oil groups, respectively. Most of the platelet clumps were adherent to attached monocytes in all groups and the number of attached monocytes were greatly reduced by the fish oil additive. Thus there were close relationships among platelet clumps, monocytes, and lesion endothelium. Numbers of attachments over nonlesion endothelium were much less than those over lesions in all dietary groups. The most surprising result was the lack of retardation of lesion growth by the fish oil additives in spite of the reduction in attached monocytes and platelet clumps. In previous studies where the high plasma cholesterol levels in the BT swine had been modestly reduced (about 25%) there had been a marked retardation of lesion growth. The current result suggests that plasma cholesterol is the major factor controlling lesion growth in this model through under milder conditions and longer observation periods other factors might become apparent. PMID- 8224114 TI - Histologic and ultrastructural features of experimental duodenal ulcers in Sprague-Dawley rats. AB - The histologic and ultrastructural features of a time sequence study of the development, evolution, and healing of acetic acid-induced experimental duodenal ulcer are presented. Duodenal ulcers produced by serosal application of acetic acid featured microvascular injury with progressive disintegration of the tips of the mucosal villi and subtotal necrosis of the duodenal wall. At 3 days ulcers transformed into a chronic state with regenerating epithelium originating from the crypts of the intact bordering mucosa extending toward the center of the ulcers. By 21 days healed ulcers were covered by distorted duodenal surface mucosa. We conclude that this reproducible and standardized model of duodenal ulcer features vascular injury as the earliest microscopic event, that ischemic necrosis leads to ulceration, and that the chronic phase bears morphologic resemblance to human duodenal ulcer. PMID- 8224115 TI - Cocaine-induced myocardial ultrastructural alterations and cardiac output responses in rats. AB - Cocaine use has been associated with profound functional and pathological myocardial responses in otherwise asymptomatic humans, yet a number of individuals appear to tolerate large doses of the drug. This study was designed to determine whether there is a relationship between the differential effects of cocaine administration on cardiovascular responses and on the development of cardiomyopathies in rats. After instrumentation for determination of cardiac output, conscious, freely moving rats were treated with cocaine (5 mg/kg) or saline intravenously twice daily for 14 days before removing the myocardium for analysis. Although most cardiovascular responses were similar, cocaine administration elicited consistent decreases in cardiac output in some rats, whereas others showed little change or an increase. While little change was evident at low magnification, electron microscopy revealed diffusely distributed myocardial lesions including focally dilated sarcoplasmic reticulum and myofibrillar derangement, early signs of mitochondrial alterations, and foci of myocardial fibrosis. The incidence of these alterations was greater in rats with a decrease in cardiac output. We also observed these lesions in a subset of rats treated with cocaine without cardiac output instrumentation. These data represent the first evidence that there is a relationship between cocaine-induced functional and pathological alterations and that rats, like humans, may be differentially sensitive to these effects. PMID- 8224116 TI - Effect of injury on mast cells of rat gastrocnemius muscle with respect to gravitational exposure. AB - The proteolytic enzyme, chymase, was used to identify mast cells in rat gastrocnemius muscles which were crush-injured or incised in order to determine if mast cells exhibited proliferation and degranulation. Some of the crush injured rats were subjected to 0 g for 14 days after injury on the Cosmos 2044 satellite to study the effects of weightlessness on the mast cell response. A variety of ground-based injured models were used, including a group of hindlimb unloaded animals acting as controls and testing the suitability of the hindlimb unloaded animal as a model for muscle healing during weightlessness. In most cases, the numbers of mast cells and their apparent size increased after injury. When mast cell degranulation was evident, the granules containing chymase often were free in the loose connective tissue and along the edge of myofibers. The mast cell response was most exaggerated in animals subjected to 0 g and least visible in the hindlimb-unloaded ones. Thus, gravitational stress may influence mast cell physiology and the hindlimb-unloaded animal may not be a good model for investigating muscle healing. PMID- 8224117 TI - Glycoprotein IIb/IIIa cross-reacting antigen in monocyte-derived macrophages from the pigeon. AB - The binding of fibrinogen to its receptor on mammalian platelets and avian thrombocytes has been extensively studied; and the receptors, composed of glycoproteins IIb and IIIa, have been characterized in both systems. Recently, monocytes have been implicated in the thrombotic complications of atherosclerosis, and in both the avian and human systems this appears to be through a procoagulant activity which leads to fibrinogen polymerization. Although fibrin polymerization by avian monocytes has been reported, the receptor for fibrinogen on these cells has not been reported previously. The present study describes the presence of glycoprotein IIb- and IIIa-like proteins in avian macrophages and correlates the localization of these glycoproteins with regions to which fibrinogen binds. Through the use of immunofluorescence light microscopy and immunogold electron microscopy in conjunction with monospecific, polyclonal antibodies, GPIIb and GPIIIa cross-reacting antigens were identified on membranes of monocyte/macrophages cultured from White Carneau pigeons. A specific concentration of the antigens was found on membrane ruffles and microvilli, sites to which FITC-labeled fibrinogen also bound. Interaction of the antibodies with pigeon macrophages was confirmed by enzyme-linked immunosorbent assays with cultured cells. Immunoblotting of membranes isolated from pigeon monocyte/macrophages identified a protein of 132,000 M(r) that was recognized by anti-GPIIb and a protein of 114,000 M(r) that was recognized by anti-GPIIIa. These pigeon monocyte glycoproteins comigrated with glycoproteins IIb and IIIa isolated from human platelets. PMID- 8224118 TI - Pulmonary immune cells: villains and confederates. PMID- 8224119 TI - Can measurements of airway responsiveness be standardized in children? PMID- 8224120 TI - How to assess long-term effects of occupational exposure. PMID- 8224121 TI - Drugs and the control of exercise-induced asthma. PMID- 8224122 TI - Lung retransplantation. PMID- 8224123 TI - Decline in FEV1 and airflow limitation related to occupational exposures in men of an urban community. AB - The purpose of this survey was to evaluate the effects of smoking and occupational exposures on the decline in forced expiratory volume in one second (FEV1), and the presence of airflow limitation (FEV1 x100/forced vital capacity (FVC) being < 65) at follow-up. A random sample of 1,933 men aged 22-54 years in Bergen, Norway, were invited into the survey. Smoking habits and measurements of FEV1 were recorded at the initial survey in 1965-1970 (n = 1,591) and at follow up in 1988-1990 (n = 951). Past or present self-reported occupational exposures to eleven airborne agents (dusts, gases, vapours and fumes) and measurements of FVC were recorded at follow-up only. The decline in FEV1 was associated (p < 0.001) with age, body height and smoking. Smoking cessation reduced the decline to the level of lifetime nonsmokers. Accelerated decline in FEV1 was observed in subjects exposed to sulphur dioxide gas and to metal fumes. The adjusted decline in FEV1 increased progressively in subjects exposed to increasing numbers of occupational agents (test for trend: p < 0.01). Airflow limitation was observed in 9.5% at follow-up, and increased with age and cigarette consumption. In this community follow-up survey in men, smoking and occupational exposures to sulphur dioxide gas, metal fumes and the numbers of specific agents were found to be important predictors for accelerated decline in FEV1. PMID- 8224124 TI - Questionnaire assessments of recent exposure to environmental tobacco smoke in relation to salivary cotinine. AB - The increasing evidence of the ill-health effects of environmental tobacco smoke (ETS) has prompted the search for accurate measures of exposure to ETS. The present study examined whether it was possible to enhance the ability of questionnaire-derived assessments of ETS exposure, to predict salivary cotinine. Salivary samples were obtained from 258 nonsmoking bank employees, who simultaneously answered questions detailing their exposure to second-hand smoke within the last three days. Exposure models were created, to take into account the number of smokers nearby, length of time in their presence, half-life of cotinine in bodily fluids, level of aversion to cigarette smoke and time of year. All models, including the consideration of intensity and duration of exposure combined, explained an equal amount of variance of log cotinine levels (approximately 16%). The weak relationship between questionnaire estimates of ETS exposure and cotinine, found in the present study, suggests that further investigation is needed to improve the assessment of recent ETS exposure. PMID- 8224125 TI - Air pollution levels, meteorological conditions and asthma symptoms. AB - We wanted to assess relations between the daily occurrence of asthma symptoms and fluctuations of air pollution concentrations and meteorological conditions. In a panel of 31 asthmatic patients residing in the town of Pitea in northern Sweden, severe symptoms of shortness of breath, wheeze, cough and phlegm were recorded in an asthma diary together with suspected causes. Sulphur dioxide, nitrogen dioxide, black smoke, relative humidity and temperature were used to evaluate the relationship to the environment. By using multivariate analyses, we found that daily variations in the particulate pollution levels, indicated by black smoke levels below the criteria limits, had significant effects on the risk of developing severe symptoms of shortness of breath. This association was stronger among 10 subjects, who had at least five incident days with severe shortness of breath. Meteorological conditions were not significant in the multivariate models. Cough and phlegm did not show significant relationships to any environmental condition that was evaluated. Only one-third of the subjects reported, at least once during the study, symptoms believed to be related to air pollutants, although we found significant correlations between the pollution levels and the frequency of pollution-related symptoms. We conclude that an association has been established for black smoke as pollutant and shortness of breath as respiratory symptom, and that in certain asthmatics, effects were occurring at lower particulate levels than suggested previously. PMID- 8224126 TI - Assessment of airway inflammation in asthmatic patients by visual endoscopic scoring systems. AB - Inflammation is a hallmark of bronchial asthma. Inflammatory cells both in bronchoalveolar lavage (BAL) and bronchial biopsies of asthmatic patients have been studied and correlated with functional or clinical parameters. We have recently attempted to assess airway inflammation by a visual endoscopic scoring system. The purpose of this study was to compare our own endoscopic scoring system with the bronchitis index previously described by Thompson and coworkers, and to determine whether these scores were correlated with clinical or functional parameters. Sixty asthmatics of variable severity (forced expiratory volume in one second (FEV1) 33-117% of predicted values) and 30 healthy volunteers were studied. The clinical severity of asthma was assessed by the clinical score as described by Aas in all of the patients, and in the last 15 patients by a daily symptom score. Beta 2-agonist consumption was recorded daily during a period of 7 days before the endoscopic procedure. During this period, circadian variation and day-to-day variation of peak expiratory flow rate (PEFR) were determined. During bronchoscopy, airway inflammation was assessed by two independent observers, prior to BAL, by visual inspection of each lobe and the lingula, and the results were quantitated using the bronchitis index and our endoscopic scoring system. Both endoscopic scores were significantly higher in asthmatics than in controls. A significant correlation was observed between the two endoscopic scores both in asthmatics and in controls. In asthmatics, a weak but significant correlation was found between both endoscopic scores and the daily symptom score, as well as the beta 2-agonist consumption.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224127 TI - Extracellular and intracellular magnesium concentrations in asthmatic patients. AB - Magnesium deficiency is associated with increased contractility of smooth muscle cells. Since contractility of bronchial smooth muscle is important in patients with asthma, magnesium deficiency could negatively influence the clinical condition. We wanted to assess whether magnesium deficiency exists in patients with asthma. Extracellular (plasma) and intracellular (erythrocytes and mononuclear leucocytes) concentrations of magnesium were determined in 20 mildly symptomatic patients with asthma and compared to 20 healthy controls. In asthmatic patients, the mean +/- SD magnesium level in plasma was 0.81 +/- 0.05 mmol.l-1, in erythrocytes 0.20 +/- 0.02 fmol.cell-1, and in mononuclear leucocytes 5.10 +/- 2.55 fmol.cell-1; these values did not differ significantly from those of the healthy controls: 0.79 +/- 0.06 mmol.l-1, 0.19 +/- 0.02 fmol.cell-1, and 4.61 +/- 1.75 fmol.cell-1, respectively. No evidence for the existence of a magnesium deficit needing chronic magnesium supplementation was, thus, found in these patients. PMID- 8224128 TI - Ventilatory pattern during bronchial histamine challenge in asthmatics. AB - We wanted to investigate whether asthmatic subjects change their ventilatory pattern consistently when forced expiratory volume in one second (FEV1) has declined by at least 20% during bronchial histamine challenge, in order to assess whether respiratory pattern analysis can be used to monitor bronchial obstruction continuously. Histamine challenge was performed twice within a four week period, in eight asthmatic teenagers. Respiratory inductive plethysmography (RIP) was used for respiratory pattern evaluation, whilst the patients breathed on a mouthpiece attached to a pneumo-tachometer (PTM) whilst wearing a noseclip (first histamine challenge), and during natural breathing (second HiCh). End-tidal carbon dioxide tension (PETCO2) was measured on both occasions. During the second histamine challenge, four of the eight patients responded with a 72% (mean) increase in minute ventilation (VE), an 80% increase in mean inspiratory flow (VI), and a 20% decrease in PETCO2. VE and VI were unchanged, or tended to decrease, among the other four patients (ventilatory nonresponders). Neither provocative dose producing a 20% fall in FEV1 (PD20) to histamine nor the magnitude of the fall in FEV1 differed between ventilatory responders and nonresponders. The ventilatory response to inhaled histamine was abolished when breathing through a PTM. Histamine induced bronchospasm is not uniformly reflected in the breathing pattern. Hyperventilation during histamine challenge might be the consequence of vagal airway receptor activation. Respiratory pattern analysis is not a feasible way to monitor bronchial obstruction during histamine challenge. PMID- 8224129 TI - Effect of furosemide on the response of laryngeal receptors to low-chloride solutions. AB - Laryngeal irritant receptors are stimulated by water and solutions lacking chloride ions, such as isotonic dextrose. It has been reported that furosemide (frusemide) reduces cough evoked by inhalation of low-chloride solutions. We studied the effect of furosemide on the response of laryngeal receptors to isotonic dextrose. Experiments were performed on nine dogs anaesthetized, spontaneously breathing through a tracheostomy, and with the upper airway functionally isolated. We recorded the activity of 13 laryngeal irritant receptors. Isotonic dextrose (4 ml) was instilled into the laryngeal lumen, before and after administration of a furosemide solution (3.75 mg.ml-1) into the upper airway. Before furosemide, dextrose increased the activity of the 13 receptors from 1.0 +/- 0.5 to 25.0 +/- 3.5 impulses (imp).s-1 (average discharge in the first 10 s of activation) and, 1-2 min after furosemide, from 0.3 +/- 0.2 to 13.4 +/- 3.2 imp.s-1; the difference between the stimulation by dextrose before and after furosemide was statistically significant. In contrast, the response to distilled water of four respiratory-modulated mechano-receptors (known to be activated by low-osmolality solutions) was not modified by furosemide. These results suggest that the furosemide-mediated inhibition of cough induced by inhalation of low-chloride solutions is, at least in part, due to the inhibitory effect of this substance on irritant receptor stimulation. PMID- 8224130 TI - Distearoyl phosphatidylglycerol liposomes improve surface and transport properties of CF mucus. AB - We have previously shown that a decreased level of phosphatidylglycerol in cystic fibrosis (CF) respiratory mucus is partly responsible for its marked adhesiveness and stickiness, which impair mucus transport, and that distearoyl phosphatidylglycerol (DSPG) was the most efficient form of phosphatidylglycerol in the enhancement of respiratory mucus clearance. The aim of our study was to analyse the effect of distearoyl phosphatidylglycerol liposomes on the transport by cough and cilia of cystic fibrosis respiratory mucus. The surface and transport properties of mucus were measured: 1) on native cystic fibrosis mucus; 2) on cystic fibrosis mucus complemented with DSPG liposomes at a non-cytotoxic concentration; and 3) on cystic fibrosis mucus complemented with water. The work of adhesion of cystic fibrosis mucus was significantly decreased by DSPG liposomes, but not by water. For mucociliary transport, the cystic fibrosis mucus was transported at a higher rate with DSPG liposomes and water compared to native cystic fibrosis mucus. The cough clearance of cystic fibrosis respiratory mucus was significantly improved in the presence of DSPG and water, but the effect was more pronounced with DSPG liposomes than with water. We conclude that the use of DSPG liposomes as a lubricating agent proves to be an interesting therapeutic approach for improving the cough and mucociliary transport in cystic fibrosis patients. PMID- 8224131 TI - Platelet-derived growth factor AA and AB dimers are present in normal human epithelial lining fluid. AB - Normal lung architecture is related to the presence of mesenchymal cells (fibroblasts and smooth muscle cells), and to the production by these cells of extracellular matrix. The turnover of mesenchymal cells is under a fine regulation due, at least in part, to the local presence of different mediators acting on their cell cycle. Since normal human alveolar macrophages obtained by bronchoalveolar lavage (BAL) spontaneously release platelet-derived growth factor (PDGF), a cytokine with chemotactic and growth activity on mesenchymal cells, we evaluated normal human epithelial lining fluid (ELF) for the presence of PDGF. Active only as a dimer, PDGF is a glycoprotein composed of two chains (A and B) and, thus, can be present in three forms: AA, AB, and BB dimers. Interestingly, normal ELF contains PDGF AA dimers, and to a lesser extent AB dimers, while no significant level of BB dimers is detected. Furthermore, ELF PDGF is biologically active and responsible for a significant part of the chemotactic activity and the "competence" growth activity for mesenchymal cells present in normal ELF. These findings suggest that ELF PDGF has a role in normal lung structure maintenance and tissue repair. PMID- 8224132 TI - Short-term in vivo exposure to cigarette-smoke increases the fluorescence in rat alveolar macrophages. AB - Alveolar macrophages (AMs) from human smokers exhibit a strong intracellular fluorescence. In order to study the mechanisms behind the fluorescence, we wanted to determine whether it could be induced by short term exposure to smoke. We therefore exposed rats for either one or five days to cigarette smoke. AMs were recovered by bronchoalveolar lavage (BAL), and their fluorescence was quantified by flow cytofluorometry. AMs from rats exposed to two cigarettes, on a single occasion, showed an increased fluorescence compared to controls. The fluorescence was further enhanced by exposing rats to two cigarettes, for five consecutive days. Larger and more granular/complex AMs were more fluorescent than smaller and less granular/complex cells. Smoke-exposed rats (five consecutive days) lavaged immediately after the exposure, had less cells in their BAL fluid than control animals. However, when rats were lavaged three smoke-free days after the exposure, the cell recovery increased, probably due to less airway obstruction. In conclusion, we found that a short-term in vivo exposure to cigarette smoke results in an increased AM fluorescence. The findings can be one explanation for the strong fluorescence seen in AMs from human smokers, possibly due to endocytosis of fluorescent material. PMID- 8224133 TI - Tobacco smoke exposure suppresses radiation-induced inflammation in the lung: a study of bronchoalveolar lavage and ultrastructural morphology in the rat. AB - Previous studies on patients with breast cancer, who received postsurgical irradiation, displayed a markedly suppressed inflammatory response in the lung of smoking patients compared to nonsmokers. The aim of the present study was to investigate further the effect of exposure to tobacco smoke on the development of irradiation-induced pneumonitis in the rat. Four groups of animals were used: controls (C); those exposed to tobacco smoke (S); those irradiated but not exposed to smoke (RNS); and those irradiated and exposed to tobacco smoke (RS). The rats were exposed to a diluted main stream of cigarette smoke, at a concentration of about 0.4 mg.l-1, in a nose-only exposure system for 1 h.day-1, 5 days.week-1 for 10 weeks. Exposure to tobacco smoke started 3 weeks before irradiation. The basal one third of both lungs was exposed to a single radiation dose of 28 Gy (6 MeV photons). All animals were killed 7 weeks after irradiation. We compared findings in bronchoalveolar lavage (BAL) and tissue morphology. The alveolar tissue showed less inflammation in the RS-group than in the RNS-group. Most strikingly, mast cells were increased one hundredfold in the lung interstitium and thirty fold in the peribronchial area in the RNS-group, whereas no increase was found in the RS-group or in the controls. The alveolar septa of the RNS-group were thickened, with occurrence of inflammatory cells and mast cells, whereas the RS-group displayed no difference as compared to the non irradiated, nonsmoking group (C).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224134 TI - Total respiratory impedance and early emphysema. AB - It can be postulated that patients in early stages of pulmonary emphysema have normal values of total respiratory resistance and reactance. The purpose of this study was to investigate whether pulmonary emphysema, detected functionally by a decrease of the single breath diffusing capacity (DLCO) by at least 25% of predicted, and an increase of the static lung compliance (CLst) by at least 50% of predicted, can be accompanied by normal values of respiratory resistance (Rrs) and reactance (Xrs), measured between 2 and 24 Hz by the forced oscillation technique. In a prospective study, we determined CLst in 26 patients, who had been selected on the basis of normal values of Rrs and Xrs, and a DLCO of less than 75% of predicted. In 17 of these patients, CLst was more than 150% of predicted. Since there were only minor abnormalities on routine lung function tests and chest X-ray, it is likely that these patients presented early emphysema. In the nine other patients, CLst was within normal limits: four suffered from interstitial lung disease; the remaining five were probably in a preliminary stage of early emphysema. In conclusion, early emphysema should systematically be considered as the first diagnosis in patients with normal values of Rrs and Xrs, and a decrease of DLCO. Onset of interstitial lung disease is a possible alternative. PMID- 8224135 TI - Inspiratory muscle performance relative to the anaerobic threshold in patients with COPD. AB - Rehabilitation programmes in chronic obstructive pulmonary disease (COPD) require exercise training above the anaerobic threshold. However, not all COPD patients develop metabolic acidosis during exercise. The hypothesis of this study was that non-exercise variables, characterizing the mechanical load on the inspiratory muscles during breathing at rest, can be used to reliably predict which patients with COPD are not able to develop metabolic acidosis during exercise. Thirty participants with COPD performed a symptom-limited cycle ergometer test. The oesophageal pressure/time index (PTIoes: the product of pressure magnitude and duration), the mean rate of pressure development during inspiration (Poes/TI), and the mean airway resistance (Raw)/maximal oesophageal pressure (Poesmax) ratio served as indices for the mechanical load on the inspiratory muscles. The oxygen uptake (VO2) at which plasma standard bicarbonate was seen to decrease from its baseline value was taken as the anaerboic threshold (AT). Mean Raw was significantly higher in those patients in whom the AT could not be detected. No other lung function parameters measured at rest allowed the accurate selection of those patients who did or did not develop exercise metabolic acidosis. On the other hand, Raw/Poesmax, PTIoes and Poes/TI were significantly different in the two patient groups. Additionally, whereas in the patient group with identifiable AT exercise hyperpnoea produced a non-linear increase of Poes/TI with respect to PTIoes above the AT, in the patient group without identifiable AT there was a linear relationship between Poes/TI and PTIoes throughout exercise. We conclude that the determination of inspiratory muscle load indices at rest may be useful in pulmonary rehabilitation programmes, for identifying those patients with COPD who do not develop exercise induced metabolic acidosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224136 TI - Respiratory dysfunction in congestive heart failure: correction after heart transplantation. AB - Severe chronic congestive heart failure (CCHF) is known to induce a restrictive ventilatory defect, with a small decrease in lung transfer factor for carbon monoxide (TLCO). The aim of the present work was to assess the reversibility of this dysfunction. We studied a group of 47 patients with CCHF, before and one year after heart transplantation. The measurements included static and dynamic lung volumes, TLCO and cardiac function. On initial evaluation, vital capacity (VC), total lung capacity (TLC) and TLCO were reduced to 76, 79 and 64% of the predicted value (% pred), respectively. Forced expiratory volume in one second (FEV1) was decreased to 69% pred, with a FEV1/VC ratio below 0.70 in 13 out of 47 patients. One year after transplantation, cardiac function had markedly improved, as shown by a normalized left ventricular ejection fraction (from 18% preoperatively to 59% postoperatively), and mean pulmonary wedge pressure (from 26 to 12 mmHg). At this time, VC (94% pred) and TLC (98% pred) were within the normal range, whereas TLCO remained low (67% pred). The FEV1/VC ratio did not change, even in the subgroup with an initial low value. Smoking habits did contribute to the low TLCO and FEV1/VC ratio. In conclusion, respiratory dysfunction induced by CCHF is reversible, with the exception of the reduction in TLCO, which probably reflects permanent changes in the lung vasculature. CCHF does not induce an obstructive ventilatory pattern. PMID- 8224137 TI - Single-lung retransplantation for late graft failure. AB - In lung or heart-lung recipients, an irreversible graft-failure may develop in connection with chronic rejection, infection or bronchial complications. A limited number of transplant-recipients have undergone a retransplantation procedure in several centres. First results are discouraging, especially in the case of early retransplantation. We decided, 3 yrs ago, to evaluate the feasibility and benefits of single-lung retransplantation in lung-transplant recipients with late graft-failure. Eight consecutive single-lung retransplantations were performed in patients with previous single-lung (n = 7), or double-lung (n = 1) transplant. Primary graft and native lung were removed in 5 and 3 patients, respectively. The delay between the two surgical procedures was 16 +/- 10 months (range 6-37 months). Three patients died within 3 months. Long term survivors experienced stable and satisfactory functional results (forced expiratory volume in one second (FEV1 63 +/- 21% predicted; range 40-103% predicted), with survival values ranging 8-20 months. One patient died of septic shock 16.5 months after retransplantation. The remaining four patients are alive. These data suggest that the retransplantation option could be considered in selected patients with late graft-failure. The final decision for retransplantation, however, is largely influenced by the current shortage of donor lungs. PMID- 8224138 TI - Pulmonary immune cells in health and disease: dendritic cells and Langerhans' cells. AB - The activation of T-lymphocytes recognizing specific antigens is a crucial and early event in the development of an immune response, but T-lymphocytes cannot respond to antigens without help of a second cell type called accessory cells or antigen-presenting cells. Studies from several groups have indicated that pulmonary dendritic cells and Langerhans' cells, like their counterparts in other tissues, are potent accessory cells, and suggest that these cells may play an extremely important role in initiating lung immune responses. The purpose of this review is to summarize current information concerning pulmonary dendritic cells and Langerhans' cells, including their origin, distribution in the lung and functional capabilities. The possible role of these cells in certain lung diseases of immune origin will also be discussed. PMID- 8224139 TI - Obliterative bronchiolitis with atypical features: CT scan and necropsy findings. AB - We describe the natural history of cryptogenic bronchiolitis obliterans in a patient followed for 24 yrs with serial pulmonary function tests and radiology. Severe, progressive airway obstruction developed, with overinflation but preservation of KCO. There was progressive hypoxaemia, which worsened on exertion; hypercapnoea was modest until late in the illness. Neither bronchodilators nor steroids were effective. The chest radiograph remained normal; CT showed irregular areas of low attenuation peripherally throughout the lungs, with Hounsfield numbers typical of emphysema, but no bullae. Postmortem studies included histology and quantitative studies of a corrosion cast of one lung. They showed marked airway narrowing at all levels, with pruning of peripheral branches, mucus plugging, and some emphysema. The case illustrates that cryptogenic bronchiolitis obliterans may be chronic and difficult to distinguish clinically or by investigation from other forms of chronic obstructive airways disease, particularly emphysema. PMID- 8224140 TI - Sole pulmonary involvement by Langerhans' cell histiocytosis in a child. AB - The case of a boy aged 4 yrs and 7 months, with isolated pulmonary involvement by Langerhans' cell histiocytosis is reported. The presentation of the disease was a sudden pneumothorax, with no previous signs of respiratory disease. The case was confirmed by S-100 and MT1 antibody staining, and was treated with pulse steroids and several pleural drainages, until the boy died after a large bilateral pneumothorax. PMID- 8224141 TI - Non-Hodgkin's lymphoma presenting as multiple cavitating pulmonary nodules. AB - Intrathoracic involvement is common in non-Hodgkin's lymphoma (NHL). We present the case of a young woman with a secondary pulmonary lymphoma (SPL) of the centroblastic subtype, with B-cell characteristics. The chest radiograph at presentation revealed an extremely rare pattern of multiple cavitating pulmonary nodules. The radiographic patterns of intrathoracic NHL are discussed. PMID- 8224142 TI - Targeting of recombinant Na+/glucose cotransporter (SGLT1) to the apical membrane. AB - A full-length Na+/glucose cotransporter cDNA (SGLT1) from rabbit intestine was subcloned into the pMAMneo mammalian expression vector and transfected by Ca2+ precipitation into Madin-Darby canine kidney (MDCK) cells. Stable MDCK transfectants isolated after clonal isolation and selection in G418 exhibited dexamethasone-inducible Na+/glucose cotransport activity under regulation of the MMTV promoter of the vector. Transfectants expressed the recombinant 75 kDa Na+/glucose cotransporter subunit as shown by Western blot, and SGLT1 mRNA as shown by Northern blot, but these were undetectable in untransfected MDCK cells. Over 93% of total recombinant transport activity was targeted to the apical membrane. This indicates that the primary amino acid sequence of SGLT1 contains the information necessary to target this transporter to the apical membrane. PMID- 8224143 TI - Changed lateral migration of phospho-LHCII in the thylakoid membrane upon acclimation of spinach to low temperatures. AB - Movement of proteins along the plant thylakoid membrane is of importance for several physiological events, such as state transitions and turnover and repair of the photosystem II complex. Such lateral migrations are impaired at low temperatures, which could contribute to the increased sensitivity of plants to photoinhibitory damages at low temperatures. The migration behaviour of phospho LHCII in thylakoid membranes isolated from cold-acclimated spinach was studied and compared to that in control membranes. The rate of migration of phospho-LHCII at low temperatures is increased 2- to 3-fold and the apparent activation energy of the migration is decreased after the cold acclimation. PMID- 8224144 TI - Negative correlation with liver cell division of a 38 kilodalton protein whose phosphorylation is enhanced by ras and G-proteins. AB - We showed earlier that the phosphorylation of a 38 kDa protein (p38) from rat liver plasma membrane is stimulated by ras or endogenous G-proteins. We have now estimated the level of expression of p38 in liver tissues from embryos at different stages of development, regenerating liver and also in tumor cell lines of hepatic origin. Our results indicate that the expression of p38 is negatively correlated with cell division. It is suggested that the phosphorylation of p38, an event which is regulated by ras proteins and G-proteins, could be involved in signal transduction processes associated with the inhibitory regulation of cell division. PMID- 8224145 TI - Effect of different endothelin receptor antagonists and of the novel non-peptide antagonist Ro 46-2005 on endothelin levels in rat plasma. AB - The goal of our study was to evaluate and compare the effects of receptor blockade with different endothelin (ET) receptor antagonists on plasma concentrations of ET-1, big ET-1 and ET-3 in conscious rats. Ro 46-2005 10 mg/kg, i.v.), a novel non-peptide antagonist of both ETA and ETB receptors, increased the concentrations of ET-1 in plasma to 200 +/- 13% of basal levels (P < 0.001). This effect was dose- and time-dependent and reached a maximum at 15 min. Ro 46 2005 had no effect on plasma concentrations of big ET-1 and only a minor effect on those of ET-3. In contrast to Ro 46-2005, the selective peptide ETA antagonists BQ-123 and FR-139317 had no effect on plasma ET-1 concentrations. The increase in plasma ET-1 concentrations by Ro 46-2005 was most likely not due to de novo synthesis, since big ET-1 levels were not increased and peak levels were reached early after compound injection, but perhaps to displacement of ET-1 from the ETB receptors. PMID- 8224146 TI - Trimeric mutant bacteriorhodopsin, D85N, shows a monophasic CD spectrum. AB - The structure of mutant bacteriorhodopsin (bR), D85N, was examined by CD and X ray diffraction at pH 7. The absorption maximum of D85N at pH 7 is located at 605 nm, which is similar to the acid-blue form of wild-type bR. D85N shows a monophasic CD band, the maximum of which is at 575 nm, although the crystalline arrangement and the trimeric structure is maintained. The acid-blue form of wild type bR shows a biphasic CD despite the similarity in absorption spectra. PMID- 8224147 TI - Expression of glutathione S-transferase P-form in primary cultured rat liver parenchymal cells by coplanar polychlorinated biphenyl congeners is suppressed by protein kinase inhibitors and dexamethasone. AB - Glutathione S-transferase P-form (GST-P, EC 2.5.1.18) mRNA was expressed by epidermal growth factor as well as by 3,4,5,3',4'-penta-chlorinated biphenyl (PenCB) in primary cultured rat liver parenchymal cells. The expression of GST-P was suppressed by inhibitors of protein kinase C and dexamethasone, an antagonist of AP-1 transcription factor activity, whereas expression of cytochrome P450IA2 by PenCB was not affected by these reagents. The AP-1 related transcription factor may be essential for the expression of GST-P by PenCB as also may be a protein kinase C type enzyme. PMID- 8224148 TI - Molecular cloning and gene organization of the mouse mitochondrial 3,2-trans enoyl-CoA isomerase. AB - 3,2-trans-enoyl-CoA isomerase (mECI, E.C. 5.3.3.8) is the key enzyme of mitochondrial beta-oxidation of unsaturated fatty acids. A mouse cDNA clone spanning the entire coding region of mECI was isolated and sequenced. Subsequently, two overlapping genomic clones containing the complete mECI gene were isolated and characterized. The mouse mECI cDNA comprises an open reading frame of 867 bp, encoding a protein of 32 kDa. The mECI gene, spanning about 15 kb, consists of seven exons. Multiple transcription starts were determined by primer extension experiments. Knowledge of the gene organization and availability of genomic clones for mouse mECI will facilitate the study of unsaturated fatty acid metabolism in normal and pathological states. PMID- 8224149 TI - Mutagenesis of human profilin locates its poly(L-proline)-binding site to a hydrophobic patch of aromatic amino acids. AB - The actin-binding protein, profilin, contains a src-homology (SH) 3-like fold (Schutt, C.E. et al., submitted), and its tight interaction with poly(L-proline) is reminiscent of the binding activity exhibited by SH3-domains. Here we demonstrate that replacements of aromatic amino acids in a hydrophobic patch on the surface of the profilin molecule abolish its poly(L-proline)-binding capacity. However, the location of this hydrophobic patch is found in another region of the molecule than that displaying structural similarities with SH3 domains. PMID- 8224150 TI - The pro sequence of lactase-phlorizin hydrolase is required for the enzyme to reach the plasma membrane. An intramolecular chaperone? AB - Various cDNAs coding for part or all of human pre-pro lactase-phlorizin hydrolase (pre-proLPH) were transfected into COS cells and the subcellular location of the lactase-related proteins assessed. Only the complete proLPH reached the plasma membrane. LPH without the pro sequence, and a construct containing the pro sequence and the lactase domain of mature LPH, accumulated intracellularly; the pro sequence with no mature domain was secreted. We conclude that the pro sequence is important for LPH to be transported to the cell surface. PMID- 8224151 TI - Functional expression of the parathyroid cell calcium receptor in Xenopus oocytes. AB - Various studies suggest the existence of a plasma membrane receptor on parathyroid cells that senses changes in the concentration of extracellular Ca2+. To test this hypothesis, Xenopus laevis oocytes were injected with poly(A)(+) enriched mRNA from bovine parathyroid cells and examined for their ability to respond to increases in the concentration of extracellular Ca2+ or other polycations. Cytosolic Ca2+ concentrations were measured indirectly by recording Cl- currents through the endogenous, cytosolic Ca(2+)-activated Cl- channel. Increasing the concentration of extracellular Ca2+ (from 0.7 to 5 mM) or Mg2+ (from 0.8 to 10 mM) elicited oscillatory increases in the Cl- current. Responses to either divalent cation were not observed in oocytes injected with water or with mRNA prepared from HL-60 cells or rat liver. Responses elicited by extracellular Mg2+ persisted when extracellular Ca2+ was reduced to low micromolar levels. La3+, Gd3+, or neomycin B also evoked oscillatory increases in the Cl- current in oocytes under conditions of low extracellular Ca2+ levels. These extracellular polycations all cause the mobilization of intracellular Ca2+ in oocytes injected with parathyroid cell mRNA like they do in intact parathyroid cells. The injection of parathyroid cell mRNA thus confers on oocytes the ability to detect and respond to changes in the concentration of extracellular polycations. The data provide compelling evidence for the existence of a cell surface Ca2+ receptor protein(s) on parathyroid cells that regulates cellular function. PMID- 8224152 TI - Stoichiometry of a Cl(-)-translocating ATPase. AB - It has been determined that Mg2+ stimulates phosphorylation while Cl- stimulates dephosphorylation of the chloride pump during its reaction sequence. The stoichiometry of ATP hydrolyzed to Cl- transported during a single cycle of the reaction sequence was ascertained. Intracellular concentrations of ATP, ADP and inorganic phosphate were determined and, coupled with an estimate of the standard free energy of hydrolysis for ATP, the operant free energy for ATP hydrolysis was calculated. Because the operating free energy of the pump is approximately one half the energy obtained from the total free energy of ATP hydrolysis, the only possible integral stoichiometries are one or, at the most, two net charges transported per cycle per ATP hydrolyzed. PMID- 8224153 TI - Petunia p34cdc2 protein kinase activity in G2/M cells obtained with a reversible cell cycle inhibitor, mimosine. AB - Protoplasts isolated from petunia leaf mesophyll are non-cycling cells mostly with 2C content. Cells regenerating from protoplast culture enter mitosis after 48 h. This experimental model is used to relate p34cdc2 kinase activity to cell cycle phase. Our results show that the histone H1 phosphorylation, and hence p34cdc2 kinase activity, peaks with G2+early M cell cycle phase. However, a trace kinase activity was already present when most cells were entering S phase. To obtain a maximum of cells in G1+S phases, the protoplast culture was treated with the rare amino acid, mimosine. Mimosine blocked plant cells derived from protoplast culture both at G1 and in early and mid S phase. Despite the increased G1+S level, p34cdc2 kinase activity did not increase. This suggests that the trace activity appearing when the majority of cells are entering S does not correspond to any putative p34cdc2 activation at G1/S transition but to the activation of the minor 4C population initially present in the leaf: the hypothesis remains that p34cdc2 kinase activity is solely related to G2+M phase in petunia. PMID- 8224154 TI - Direct gene transfer into mouse diaphragm. AB - Direct gene transfer into skeletal muscle is a potential therapeutic strategy for inherited primary myopathies such as Duchenne muscular dystrophy (DMD). In order to affect the life-expectancy of these patients, it will be necessary to carry out gene therapy on the diaphragm. To this end, we report efficient introduction of pure recombinant plasmid DNA into the mouse diaphragm, without causing significant damage. Application of this approach to the diaphragm of the mdx mouse will provide information on the potential usefulness of gene therapy for the treatment of DMD patients. PMID- 8224155 TI - The primary structure of inhibitor of cysteine proteinases from potato. AB - The complete amino acid sequence of the cysteine proteinase inhibitor from potato tubers was determined. The inhibitor is a single-chain protein having 180 amino acid residues. Its primary structure was elucidated by automatic degradation of the intact protein and sequence analysis of peptides generated by CNBr, trypsin and glycyl endopeptidase. A search through the protein sequence database showed homology to other plant proteinase inhibitors of different specificities and non inhibitory proteins of M(r) around 20,000. On the basis of sequence homology, prediction of secondary structure and fold compatibility, based on a 3D-1D score to the three-dimensional profile of Erythrina caffra trypsin inhibitor, we suggest that the potato cysteine proteinase inhibitor belongs to the superfamily of proteins that have the same pattern of three-dimensional structure as soybean trypsin inhibitor. This superfamily would therefore include proteins that inhibit three different classes of proteinases-serine, cysteine and aspartic proteinases. PMID- 8224156 TI - Free hydroxyl radicals are formed on reaction between the neutrophil-derived species superoxide anion and hypochlorous acid. AB - Superoxide anion reacts with hypochlorous acid to yield free hydroxyl radicals, as shown by the hydroxylation of benzoate. This reaction is analogous to the Haber-Weiss reaction but in the absence of metal ions is at least six orders of magnitude faster. PMID- 8224157 TI - Intraorganellar localization of CoASH-independent phytanic acid oxidation in human liver peroxisomes. AB - In human tissues phytanic acid is alpha-oxidized to pristanic acid in peroxisomes. Studies of the intraorganellar site of alpha-oxidation of [1 14C]phytanic acid to pristanic acid in peroxisomes isolated from human liver demonstrate that phytanoyl-CoA ligase is present in the peroxisomal membrane and that the enzyme system for alpha-oxidation of phytanic acid to pristanic acid is in the peroxisomal matrix. In contrast to the beta-oxidation system for fatty acids, the substrate for alpha-oxidation is free phytanic acid. The studies described in this manuscript report a novel fatty acid oxidation system where the substrate for the enzyme system is free fatty acid; however, phytanoyl-CoA ligase regulates the alpha-oxidation of phytanic acid at the organellar (peroxisomal) level. PMID- 8224158 TI - Sucrose-phosphate synthase phosphatase, a type 2A protein phosphatase, changes its sensitivity towards inhibition by inorganic phosphate in spinach leaves. AB - The activity of a type 2A protein phosphatase from spinach leaves was monitored using phosphorylated sucrose-phosphate synthase (SPS) as a substrate. After partial purification the overall activities of sucrose-phosphate synthase phosphatase (SPS-P) recovered from leaves harvested in the dark and in the light did not vary. However, SPS-P preparations from darkened leaves were more strongly inhibited by inorganic phosphate and certain phosphorylated compounds than preparations from illuminated or mannose fed leaves. We conclude, that activation of SPS involves an interconversion of multiple forms of SPS-P activity. PMID- 8224159 TI - Catabolite inactivation of the yeast maltose transporter is due to proteolysis. AB - The maltose transport capacity of fermenting Saccharomyces cerevisiae rapidly decreases when protein synthesis is impaired. Using polyclonal antibodies against a recombinant maltose transporter-protein we measured the cellular content of the transporter along this inactivation process. Loss of transport capacity was paralleled by a decrease of cross-reacting material which suggests degradation of the transporter. We also show that in ammonium-starved cells the half-life of the maltose transporter is 1.3 h during catabolism of glucose and > 15 h during catabolism of ethanol. PMID- 8224160 TI - Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae. AB - Autophagy in the yeast is similar to that in mammalian cells. A mutant designated as apg1 (autophagy) defective in accumulation of autophagic bodies in the vacuoles was isolated by selection using a light microscope from a mutagenized proteinase-deficient strain. In the apg1 strain, which has normal vacuolar proteinases, nitrogen starvation did not induce protein degradation. The apg1 mutant lost its viability faster than wild-type cells during nitrogen starvation. By using the loss of viability as a first screening test, 75 other apg mutants were selected. These apg mutants including apg1 fell into 15 complementation groups. Genetic analyses of representative apg mutants revealed that they all had single recessive chromosomal mutations. Strains with each apg mutation were defective in protein degradation in the vacuoles induced by nitrogen starvation and homozygous diploids for each apg mutation did not sporulate. These results on the apg mutants suggest that autophagy via autophagic bodies is indispensable for protein degradation in the vacuoles under starvation conditions, and that at least 15 APG genes are involved in autophagy in yeast. PMID- 8224161 TI - The protein kinase mos activates MAP kinase kinase in vitro and stimulates the MAP kinase pathway in mammalian somatic cells in vivo. AB - The mos protooncogene encodes a serine/threonine protein kinase that is only expressed at significant levels in germ cells. Recombinant malE-mos protein (Xenopus mos protooncogene fused in frame to the maltose binding protein of E. coli) activates MAP kinase in cell-free extracts prepared from Xenopus oocytes and eggs. Here we show that malE-mos immunoprecipitates from Xenopus extracts phosphorylate and activate MAP kinase kinase in vitro, indicating that mos can function as a MAP kinase kinase kinase. Moreover, ectopic expression of mos in mammalian somatic cells, that lack any endogenous mos protein, triggers the activation of MAP kinase in vivo. These results identify the mos protooncogene as a direct activator of the MAP kinase pathway, with the potential to activate this kinase cascade even in cells where normally there is no expression of mos. PMID- 8224162 TI - Differential inhibition by cyclosporins of primary-active ATP-dependent transporters in the hepatocyte canalicular membrane. AB - The distinct ATP-dependent transporters for taurocholate, leukotriene C4, and daunorubicin, studied in rat liver canalicular membrane vesicles, are sensitive to inhibition by cyclosporin A and its non-immunosuppressive analog PSC 833. Ki values for cyclosporin A were 0.2, 3.4 and 1.5 microM for the transport of taurocholate, leukotriene C4, and daunorubicin, respectively. The corresponding Ki values for PSC 833 were 0.6, 29, and 0.3 microM. Both inhibitors were competitive with respect to the three substrates. The cyclosporins serve as new and potent tools to interfere with different potency with the distinct ATP dependent export carriers in the hepatocyte canalicular membrane. PMID- 8224163 TI - Kinetic and structural characterization of an intermediate in the biomineralization of bacterioferritin. AB - The mechanism by which iron-storage proteins take up and oxidise iron(II) is not understood. We show by rapid-kinetic and EPR measurements that iron uptake, in vitro, by a bacterial iron-storage protein, bacterioferritin, involves at least three kinetically distinguishable phases: phase 1, the binding of Fe(II) ions, probably at a dimeric iron ferroxidase centre; phase 2, oxidation of the Fe(II) dimer and production of mononuclear Fe(III); and phase 3, iron core formation. PMID- 8224164 TI - The gag precursor contains a specific HIV-1 protease cleavage site between the NC (P7) and P1 proteins. AB - The predicted protease cleavage site (p7/p1; [J. Virol. 66 (1992) 1856-1865]) within the nucleocapsid precursor protein (p15) of human immunodeficiency virus, type 1, was confirmed using an in vitro assay employing recombinant HIV-1 protease and a chemically synthesized 72 amino acid polypeptide containing the p7 and p1 protein domains of the native gag polyprotein. The cleavage occurred between amino acid 55 (N) and amino acid 56 (F) of the polypeptide, as determined by N-terminal sequencing. The hydrolysis was optimal at pH 6.0 and at high salt concentration. The kinetic parameters Km, kcat and kcat/Km were 99 microM (+/- 8), 0.152 s-1 (+/- 0.002) and 1.56 mM-1.s-1 (+/- 0.11), respectively. Reconstituted as well as denatured polypeptides were cleaved at approximately the same rate, demonstrating that the conformation of the p7 protein, as a result of the Zn(2+)-binding, had no significant effect on the rate of hydrolysis of the p7/p1 cleavage. PMID- 8224165 TI - Cloning and characterization of the rat 5-HT5B receptor. Evidence that the 5-HT5B receptor couples to a G protein in mammalian cell membranes. AB - A gene encoding a novel G protein-coupled 5-hydroxytryptamine (5-HT) receptor, termed 5-HT5B, was cloned. The ligand binding profile of this receptor is distinct from that of other cloned 5-HT receptors. The 5-HT5B receptor couples to a G protein in COS1 cell membranes; however, activation of the 5-HT5B receptor does not appear to alter either cAMP accumulation or phosphoinositide turnover in a variety of fibroblast cell lines. In the rat brain, 5-HT5B gene expression occurs predominantly in the medial habenulae and hippocampal CA1 cells of the adult. Little expression is seen during embryonic development. PMID- 8224166 TI - Potassium regulates plasma testosterone and renal ornithine decarboxylase in mice. AB - Potassium deficiency produced different effects in the kidney of male or female mice. While in female, potassium deficiency caused a marked renal hypertrophy with no significant changes in testosterone-regulated enzymes, such as ornithine decarboxylase and beta-glucuronidase, in the male the same treatment provoked a marked fall of these enzymes owing to a dramatic decrease in plasma testosterone. Potassium replenishment restored plasma testosterone and renal enzymatic activities. These results show for the first time, that potassium modulates circulating testosterone and suggest that this cation could exert an important regulatory role in controlling androgen actions. PMID- 8224167 TI - Association constants of anti-hapten monoclonal IgG1 with mouse Fc gamma RII in the presence and absence of hapten. AB - The binding of five different mouse monoclonal anti-dinitrophenyl (DNP) IgGl monomers, in the presence and absence of DNP-glycine, to Fc gamma RII on the mouse macrophage-like cell line J774 has been investigated. Membrane association constants were estimated using a competitive radioimmunoassay in which the ability of the IgG1 monomers to block the binding of 125I-labelled Fabs of the anti-Fc gamma RII monoclonal antibody 2.4G2 was monitored. The IgG1-Fc gamma RII association constants ranged from (8 +/- 1) x 10(4) M-1 to (6 +/- 2) x 10(5) M-1. No large differences in the IgG1-Fc gamma RII association constants were measured for different IgG1 antibodies or for any single IgG1 antibody in the presence and absence of DNP-glycine. PMID- 8224168 TI - The primary structure of carboxypeptidase S1 from Penicillium janthinellum. AB - The complete amino acid sequence of carboxypeptidase S1 from Penicillium janthinellium has been determined by N-terminal sequencing of the reduced and vinylpyridinated protein and of peptides obtained by cleaved with cyanogen bromide, iodosobenzoic acid, hydroxylamine, endoproteinase LysC, endoproteinase AspN and Glu-specific proteinase from B. licheniformis. The enzyme consists of a single peptide chain of 433 amino acid residues and contains 9 half-cystine residues and one glycosylated asparagine residue. A comparison to other carboxypeptidases shows that the enzyme is homologous to carboxypeptidase-Y and carboxypeptidase-MIII from malt. Specificity and binding of substrates are discussed from a three-dimensional model based on the known structure of carboxypeptidase-Y from Saccharomyces cereviciae and carboxypeptidase II from wheat. PMID- 8224169 TI - Two proteins involved in kinetoplast compaction. AB - The kinetoplast is the genome of the single mitochondrion of trypanosomatid Protozoa, and contains up to 30% of total cellular DNA in a network of catenated AT-rich rings. EM studies show that the kinetoplast is organized into a compact, disc-shaped structure in vivo, but little is known about proteins involved in its architecture. Defining such proteins would be useful to understand the molecular biology of this unusual organelle and to design compounds to contain parasite growth. We show here that two proteins, p1 and p2 of M(r) approximately 22 and approximately 21 kDa, respectively, from the trypanosomatid Crithidia fasciculata can compact kDNA networks efficiently in vitro, the first such demonstration with purified trypanosome proteins. We show that these proteins are localized exclusively in the parasite's kinetoplast. Our data thus define two proteins potentially involved in kinetoplast organization in vivo. PMID- 8224170 TI - Cloning of CRP2, a novel member of the cysteine-rich protein family with two repeats of an unusual LIM/double zinc-finger motif. AB - The cDNA coding for a novel member of the cysteine-rich protein family was isolated from a rat brain cDNA library. It encodes a protein, denoted cysteine rich protein 2 (CRP2), of 208 amino acid residues containing two tandem repeats of an unusual LIM/double zinc-finger-like motif. The ubiquitous tissue distribution and high level of expression of CRP2 mRNA suggest an important role for CRP2 in cell functions. PMID- 8224171 TI - rab3-peptide stimulates exocytosis from mast cells via a pertussis toxin sensitive mechanism. AB - Synthetic peptides of the putative effector domain of members of the rab3 gene family of small GTP-binding proteins have been shown to have potent actions on vesicular transport and exocytosis [1,2]. Here, we use similar rab3-effector domain peptides to study their role in intracellular signalling in mast cells. We find that rab3-like peptides stimulate exocytosis and decrease cyclic 3',5'-AMP levels in these cells when applied extracellularly. Cells pretreated with pertussis toxin (PtX) to selectively uncouple alpha i/alpha o type G proteins from their biological activators, however, did not respond to rab3 peptides. rab3 like peptides also induce a Ca2+ transient in mast cells. These observations provide evidence for functional coupling between an effector domain peptide sequence of rab3 protein and a PtX-sensitive G protein substrate. PMID- 8224172 TI - Hepatocyte growth factor-like protein is identical to macrophage stimulating protein. AB - Although the hepatocyte growth factor-like protein (HLP) shares a 50% homology with the hepatocyte growth factor, the biological function of HLP has remained unknown. Addition of conditioned medium of COS-7 cells transfected with the expression plasmid for HLP cDNA to cultures of resident peritoneal macrophages induced specific activation of macrophages, and the factor which stimulates macrophages was purified from the conditioned medium. The purified protein showed M(r) of 85 kDa on SDS-PAGE, and this M(r) is in agreement with that of macrophage stimulating protein (MSP) previously purified from human serum, as well as with the predicted M(r) of HLP. Amino acid composition of the purified protein coincided with the compositions of human HLP and MSP. Together with the finding that the partial amino acid sequences of MSP are highly homologous to that of HLP, we conclude that the biological function of HLP is to activate macrophages and that HLP and MSP are identical molecules. PMID- 8224173 TI - Characterisation of protein kinase C isoforms and enzymic activity from the alpha T3-1 gonadotroph-derived cell line. AB - Western blots of alpha T3-1 cell extracts were immunostained with antibodies specific for various protein kinase C (PKC) isoforms. These revealed the presence of PKC types alpha, epsilon and zeta, but beta, gamma, delta and eta were not detected. The potency with which partially-purified cytosolic PKC from alpha T3-1 cells was activated by phorbol 12,13-dibutyrate (PDBu), mezerein and 1,2 dioctanoyl-sn-glycerol was assessed in the presence and absence of Ca2+. The inhibitors staurosporine, K252a, H7, GF109203X and Ro 31-8220 were tested on basal activity, PDBu-induced activity and Ca(2+) + PDBu-induced kinase activity. Each inhibitor showed distinct differences in their IC50 values under the three conditions, suggesting that these inhibitors may exhibit different potencies on the PKC isoforms present in alpha T3-1 cells. Although histone IIIs was used as the phosphate acceptor for most of these experiments, the efficiency of alpha, epsilon and zeta peptide and GS peptide substrates were also determined, with epsilon peptide giving the greatest activity in the presence of PDBu or Ca2+. Each substrate displayed a different pattern of activation under the conditions tested. Overall, the findings suggest that 3 or more PKC isoforms with varying specificities are present in gonadotroph-derived alpha T3-1 cells and that the contribution of each isoform should be considered when these cells are used in models of anterior pituitary cell function where PKC is involved. PMID- 8224174 TI - Effects of dexamethasone on L-type calcium currents in the A7r5 smooth muscle derived cell line. AB - Patch clamp experiments were used to characterize the effect of dexamethasone on calcium currents in A7r5 cells. Pretreatment for 48 h with 200 nM dexamethasone did not affect the single channel conductance, the voltage dependence of channel opening, or the voltage-dependent inactivation of L-type channels. However, dexamethasone caused an approximately 2-fold increase in the amplitude of L-type calcium currents in 5 out of 9 experiments, suggesting an increase in the number of active channels. The effect of dexamethasone appeared to be greatest on batches of cells with low control current density. The amplitude of T-type calcium current was not affected by dexamethasone. PMID- 8224175 TI - H(+)-coupled (Na(+)-independent) proline transport in human intestinal (Caco-2) epithelial cell monolayers. AB - Previously, absorption of L-proline across the apical membrane of the intestinal enterocyte has been attributed to transport via the Na(+)-dependent Imino system. However, net (absorptive) transport of proline across intact Caco-2 cell monolayers was enhanced by acidification of the apical environment, under both Na(+)-containing and Na(+)-free conditions. This Na(+)-independent pH-dependent proline flux was associated with H+ flow across the apical membrane as determined by continuous measurement of intracellular pH. H+/proline symport was associated with an inward Isc in voltage-clamped Caco-2 epithelial layers demonstrating the electrogenic nature of this transport process. In conclusion Caco-2 cells possess an apically-localised, Na(+)-independent, electrogenic H+/imino acid transporter which may play an important role in intestinal proline absorption. PMID- 8224176 TI - Uterine cells other than stromal decidual cells are required for 1,25 dihydroxyvitamin D3 production during early human pregnancy. AB - Human decidual cells are known to produce 1,25-(OH)2D3 at the end of pregnancy, the present study evaluates this capacity, and the part played by stromal decidual cells, in early pregnancy. Cells were obtained from nine human decidua by aspiration or curettage during early pregnancy (7-10 weeks), separated on Ficoll-Paque and plastic adherence, and incubated for 1 h with 25-(OH)D3. Incubation medium and cells were extracted and chromatographed on two successive HPLC systems. The cells examined were of both physiological and pathological (ectopic pregnancy) origin. Endometrial cells obtained in four non-pregnant situations (myomas) were also studied to determine whether the 1,25-(OH)2D3 synthesis by the uterus is associated with the appearance of decidual cells. Results show that human decidual cells from early pregnancy convert 25(OH)D3 (2.5 nM or 2.5 microM) into a metabolite with the physicochemical characteristics of synthetic 1,25-(OH)2D3. This ability is shared by cells isolated during early pregnancy, whether physiological or ectopic (tubal pregnancy). Non-adherent cells, which include mainly stromal decidual cells, are less able to produce 1,25 (OH)2D3 than are the adherent cells, suggesting that macrophages, granulocytes or as yet unidentified cell types are required for the 1,25-(OH)2D3 production by decidual tissue during early human pregnancy. In addition, one out of four experiments with non-pregnant endometrial cells could produce 1,25-(OH)2D3 suggesting that, although not the rule in the non-pregnant state, in vitro production of 1,25-(OH)2D3 by uterine cells can be found in the absence of decidual cells. PMID- 8224177 TI - Effects of a single intrastrand d(GpG) platinum adduct on the strand separating activity of the Escherichia coli proteins RecB and RecA. AB - RecB and RecA proteins play key roles in the process of DNA recombination in Escherichia coli and both possess DNA unwinding activities which can displace short regions of duplex DNA in an ATP-dependent manner in vitro. We have examined the effect of the most abundant DNA adduct caused by the chemotherapeutic agent cis-diamminedichloroplatinum(II) on those activities. For this purpose, we have constructed a partially duplex synthetic oligonucleotide containing the intrastrand d(GpG) crosslink positioned at a specific site. We report here that both the DNA strand separating and DNA-dependent ATPase activities of the RecB protein are inhibited by the d(GpG) cis-DDP adduct. In contrast, neither the unwinding nor the ATPase activities of RecA protein appear to be perturbed by this lesion. PMID- 8224178 TI - A novel peptide designed for sensitization of terbium (III) luminescence. AB - Several synthetic peptides, modelled from a Ca(2+)-binding loop of the EF-hand family of proteins, were prepared containing cysteine residues. The peptide, GDKNADGFICFEEL, was labelled covalently at the cysteine residue (loop position 9) with iodoacetamidosalicylic acid. This novel conjugate is a metal-binding loop containing a salicylic acid side chain that could not only chelate Tb3+ in conjunction with the other chelating groups in the sequence, but could also sensitize Tb3+ luminescence. The loop had a high Tb3+ affinity, with stoichiometric binding observed under experimental conditions. The luminescence from the Tb(3+)-peptide complex was more than 10-fold greater than the luminescence reported from a related peptide which contained Trp as the Tb3+ donor at loop position 7. This peptide has significant potential for use in lanthanide-based time-resolved luminescence immunoassays. PMID- 8224179 TI - Structural relationship of streptavidin to the calycin protein superfamily. AB - Streptavidin is a binding protein, from the bacteria Streptomyces avidinii, with remarkable affinity for the vitamin biotin. The lipocalins and the fatty acid binding proteins (FABPs), are two other protein families which also act by binding small hydrophobic molecules. Within a similar overall folding pattern (a beta-barrel with a repeated +1 topology), large parts of the lipocalin, FABP, and streptavidin molecules can be structurally equivalenced. The first structurally conserved region within the three-dimensional alignment, or common core, characteristic of the three groups corresponds to an unusual structural feature (a short 3(10) helix leading into a beta-strand, the first of the barrel), conserved in both its conformation and its location within their folds, which also displays characteristic sequence conservation. These similarities of structure and sequence suggest that all three families form part of a larger group: the calycin structural superfamily. PMID- 8224180 TI - Does the HIV Nef protein mimic the MHC? AB - The sequence of the HIV Nef protein has no significant homology to other proteins in the SwissProt database, and experimental data concerning its function are sparse and contradictory. Using a novel protein sequence comparison method, we find similarities between different Nef sequences and the alpha chain of human MHC class I proteins. The possible biological implications of this finding are discussed. PMID- 8224181 TI - Cooperative face-to-face and edge-to-face aromatic interactions of tryptophan indole ring with N7-quarternized guanine and neutral cytosine bases. AB - In order to investigate the effect of cytosine base upon the stacking interaction of N7-quarternized guanine base with tryptophan indole ring, the X-ray crystal structure of a 1:1 complex of model compounds 1 and 2 was carried out. Contrary to the expectation of the interaction of both molecules in aqueous solution, the crystal structure showed the first example of the simultaneous recognition of the Trp indole ring by guanine and cytosine bases by the coupling of the face-to-face and edge-to-face aromatic interactions, respectively. PMID- 8224182 TI - Verification of the interaction between peptide T and CD4 using surface plasmon resonance. AB - Peptide T is currently in phase II clinical trials for the treatment of AIDS associated dementia. Its putative mode of action is inhibition of binding of the HIV envelope protein (gp120) to its cellular receptor (CD4), thus preventing viral infectivity and gp120-induced neuronal toxicity. However, a number of reports have appeared in the literature which have failed to observe any inhibitory activity of Peptide T on CD4-gp120 binding, thus casting doubt on this hypothesis. This study uses a novel biosensor technique to demonstrate that Peptide T does bind to CD4 and that this binding can be specifically inhibited by an anti-CD4 monoclonal antibody. A detailed analysis of the kinetics of the interaction is presented. PMID- 8224183 TI - Plasminogen-binding protein associated with the plasma membrane of cultured embryonic rat neocortical neurons. AB - To investigate the receptor-like molecule(s) for plasminogen (PGn) on the neuronal surface, the properties of binding of PGn to the plasma membrane of cultured embryonic rat neocortical neurons were investigated. [125I]PGn was found to specifically bind to the plasma membrane depending on the incubation temperature and time. The binding was also affected strongly by ionic strength and slightly by Ca2+. Furthermore, ligand blotting analysis revealed that [125I]PGn binds to a major protein with an apparent molecular weight of 45 kDa among plasma membrane proteins. These results suggest that the 45-kDa protein is a PGn receptor-like molecule on the neuronal surface. PMID- 8224184 TI - Overexpression of protein kinase C-epsilon enhances the stimulatory effect of ethanol on phospholipase C-mediated hydrolysis of phosphatidylethanolamine in NIH 3T3 fibroblasts. AB - Previously, ethanol and the protein kinase C (PKC) activators phorbol 12 myristate 13-acetate (PMA) and bombesin were shown to synergistically stimulate phospholipase C (PLC)-mediated hydrolysis of phosphatidylethanolamine (PtdEtn) in NIH 3T3 fibroblasts. Here we used fibroblasts overexpressing PKC-epsilon 15-fold to examine the possible role of this enzyme in the regulation of PtdEtn hydrolysis by ethanol. Overexpressed PKC-epsilon (i) greatly enhanced the stimulatory effects of ethanol (37.5-150 mM) on PLC-mediated PtdEtn hydrolysis, and (ii) eliminated the need for the co-presence of a PKC activator for maximal (3.3-fold) stimulation of PLC by 150 mM ethanol. Results suggest that PKC-epsilon is a potential positive regulator of the PtdEtn-hydrolyzing PLC activity, and that the functional interaction between PKC-epsilon and PLC is facilitated by ethanol. PMID- 8224185 TI - Transcriptional regulation of the isocitrate lyase encoding gene in Saccharomyces cerevisiae. AB - In this work, we studied the transcriptional regulation of isocitrate lyase synthesis. In Northern blot analyses we first showed that the steady-state ICL1 mRNA levels depend on the carbon source used for growth. In addition, we determined the kinetics of transcriptional repression upon a shift of ethanol grown cells to glucose and of the induction when cells were transferred from glucose to ethanol. By deletion analyses as well as by studying the influence on expression of different fragments cloned into the heterologous CYC1 promoter lacking its own UAS sequences, we defined UAS and URS elements in the ICL1 promoter. A region mediating the control by CAT3, a gene also involved in the control of expression of other genes subject to carbon catabolite repression, was found to overlap with one of these UAS elements. PMID- 8224186 TI - Conventional protein kinase C isoforms are not essential for cellular proliferation of a T cell lymphoma line. AB - The role of conventional protein kinase C (PKC) isoforms in the proliferation of a T cell lymphoma line was investigated, PKC isozyme-deficient cell lines were generated from HuT 78 which expresses PKC isoforms alpha, beta, epsilon and zeta. The first of these, K-4, expresses PKC alpha, epsilon and zeta but not beta. L-2, a mutant of K-4, expresses PKC alpha and zeta but expresses neither PKC alpha, beta, gamma, epsilon nor zeta when cultured continuously in PMA (L-2/PMA). All four cell lines continued to proliferate although at reduced rates in the presence of the PKC inhibitors staurosporine and H-7. K-4 and HuT 78 undergo growth arrest when activated with PMA (rate of proliferation decreases by 76 and 95%, respectively). While PMA-induced growth arrest occurs in L-2 cells which possess PKC alpha and zeta, PMA-induced growth arrest does not occur in L-2/PMA which is deficient in these isoforms. As these cell lines continue to proliferate in the presence of PKC inhibitors and since proliferation occurs in the absence of PKC isoforms (L-2/PMA), the results suggest that neither conventional PKC nor the epsilon and zeta isoforms are essential for proliferation of these cells. These data further suggest that PKC alpha and/or zeta may be involved in PMA induced growth arrest. PMID- 8224187 TI - Triplet repeat sequences in human DNA can be detected by hybridization to a synthetic (5'-CGG-3')17 oligodeoxyribonucleotide. AB - The seemingly autonomous amplification of naturally occurring triplet repeat sequences in the human genome has been implicated in the causation of human genetic disease, such as the fragile X (Martin-Bell) syndrome, myotonic dystrophy (Curshmann-Steinert), spinal and bulbar muscular atrophy (Kennedy's disease) and Huntington's disease. The molecular mechanisms underlying these triplet amplifications are still unknown. We demonstrate here that a synthetic (CGG)17 oligodeoxyribonucleotide can be utilized as hybridization probe to visualize some of the triplet repeats in the human genome. This technique may help in studies aimed at the elucidation of the amplification mechanism. PMID- 8224188 TI - Extensive 1H NMR resonance assignment of proteins using natural abundance gradient-enhanced 13C-1H correlation spectroscopy. AB - The reliability and completeness of 1H NMR resonance assignment can be improved by the use of 13C-1H HSQC correlation spectra on unlabelled protein samples using pulsed field gradients. This technique is illustrated on a 5.2 mM sample of the 79 residue Desulfovibrio vulgaris ferrocytochrome c553. Protons attached to the same carbon can be unambiguously paired in a HSQC spectrum. Contrary to 1H, most amino acids exhibit characteristic 13C chemical shift ranges, which can be used for 13C assignment. This technique is especially useful for long side chain residues, such as Gln, Glu, Lys, Arg. PMID- 8224189 TI - The three-dimensional structure of the colicin E3 immunity protein by distance geometry calculation. AB - The three-dimensional solution structure of the colicin E3 immunity protein (84 residues) was determined by distance geometry calculations. The hydrophilic side of a four-stranded antiparallel beta-sheet constitutes a part of the surface of the protein, and two loops lie on the hydrophobic side of the sheet. All the three specificity-determining residues, which are included in the center of the beta-sheet, display their side groups on the protein surface. PMID- 8224190 TI - Escherichia coli single-stranded DNA-binding protein alters the structure of intramolecular triplexes in plasmids. AB - The ability of the Escherichia coli single-stranded DNA-binding protein (SSB) to recognize structural features associated with intramolecular triplex formation in oligopurine.oligopyrimidine (pur.pyr) inserts in recombinant plasmids was evaluated. The SSB protein binds to supercoiled plasmids and causes a site preferential increase in OsO4 reactivity of the pyrimidine strand involved in the formation of the Hy-3 isomer of the triplex structure. The E. coli RecA protein showed no reaction with triplexes in similar studies. This behavior is consistent with SSB-mediated unpairing of the H-DNA-forming region. PMID- 8224191 TI - Heterocomplexes of tick-borne encephalitis structural and non-structural proteins. AB - The existence of the complex of structural and non-structural proteins in the tick-borne encephalitis (TBE) virus is shown. The complex was isolated from virus containing cultural medium by immunoaffinity chromatography on monoclonal antibodies (MAbs). By enzyme immunoassay and immunoblotting with the use of appropriate MAbs it was demonstrated that this complex consists of structural (protein E), and non-structural (NS1) glycoproteins. Also, the trimer E-NS1-NS3 can be isolated. It is proposed that this trimer is the viral replicative complex. PMID- 8224192 TI - Acidic pH increases the oxidation of LDL by macrophages. AB - We have investigated the effect of pH on LDL oxidation by macrophages (in the presence of iron ions), using a modification of Hanks' balanced salt solution. Increasing the acidity of the medium greatly increased the oxidation of the LDL by the macrophages as measured by thiobarbituric acid-reactive substances or increased uptake and degradation by a second set of macrophages. The rate of oxidation of LDL by iron ions alone, measured in terms of conjugated dienes, was also increased greatly even at mildly acidic pH. It is quite possible that atherosclerotic lesions have an acidic extracellular pH, particularly in the vicinity of macrophages, and the observation that LDL oxidation by macrophages is increased at acidic pH may therefore help to explain why atherosclerotic lesions are apparently one of the very few sites in the body where LDL oxidation occurs. PMID- 8224193 TI - Expression of Saccharomyces cerevisiae inorganic pyrophosphatase in Escherichia coli. AB - A segment of DNA encoding Saccharomyces cerevisiae inorganic pyrophosphatase (ppa gene) was amplified by the polymerase chain reaction. The pSCH1 and pSCB6 plasmids containing the ppa gene were obtained. Transformation of the E. coli BL21 strain with the resulting recombinant plasmids and selection of clones having extremely high expression of inorganic pyrophosphatase (PPase) were carried out. Superproduction of recombinant S. cerevisiae PPase up to 50% of the total bacterial protein was achieved. The enzyme was readily obtained and purified to homogeneity with the use of a simple purification technique. This work is the first description of S. cerevisiae PPase superproducer creation. PMID- 8224194 TI - Rapid pressure driven exocytosis-endocytosis cycle in a single plant cell. Capacitance measurements in aleurone protoplasts. AB - The whole-cell patch-clamp technique has been used to increase intracellular pressure via the pipette while monitoring changes in membrane capacitance (related to membrane area) in single aleurone protoplasts. Increased pressure increased membrane capacitance and diameter, upon the release of pressure both parameters returned to resting levels. Pressure also reversibly increased membrane conductance. Comparison between diameter and capacitance shows that the capacitance changes are due to changes in cell surface area. These results show that pressure can rapidly alter the balance between exo- and endocytosis. Pressure-induced changes in conductance are suggested to be due to insertion/withdrawal of channels with the exo-/endocytosed membrane. PMID- 8224195 TI - Insulin activates myelin basic protein (p42 MAP) kinase by a protein kinase C independent pathway in rat adipocytes. Dissociation from glucose transport. AB - Myelin basic protein kinase (MBPK) activity of rat adipocytes was measured directly or in gels after purification of p42 microtubule-associated protein kinase (MAPK). Insulin and phorbol esters provoked 2- to 3-fold increases in MBPK/MAPK activity within 5-10 min. Whereas phorbol ester effects were blocked by protein kinase C (PKC) depletion or inhibition, insulin effects were fully intact, indicating that insulin activates MBPK/MAPK independently of PKC. In contrast, PKC depletion or inhibition markedly inhibited insulin effects on [3H]2 deoxyglucose uptake, suggesting that this effect requires PKC, rather than a factor within the ras/MAPK cascade. PMID- 8224196 TI - Endogenous nitric oxide induced by interleukin-1 beta in rat islets of Langerhans and HIT-T15 cells causes significant DNA damage as measured by the 'comet' assay. AB - We have used the comet assay (single cell gel electrophoresis) to measure nitric oxide-induced DNA damage in rat islets of Langerhans and insulin-containing HIT T15 cells. Damage was induced following treatment with the nitric oxide donor SIN 1, which also releases superoxide, but was not reduced by exogenous superoxide dismutase, suggesting that nitric oxide itself, rather than superoxide or peroxynitrite may be the active species. The DNA damaging effect of nitric oxide was easily detectable at the earliest time point tested (15 min). Damage also resulted following induction of nitric oxide synthase by the cytokine interleukin 1 beta in both islets and HIT-T15 cells and was prevented by replacing the substrate, arginine, with nitromonomethyl arginine. Thus intracellular levels of nitric oxide generated by interleukin-1 beta-induced nitric oxide synthase were sufficient to cause DNA damage in islet cells and HIT-T15 cells. PMID- 8224197 TI - Physical and chemical characterization of the oligomerization state of the Aeromonas hydrophila lipase/acyltransferase. AB - Aeromonas glycerophospholipid:cholesterol acyl transferase undergoes a conformational transition upon activation by treatment with trypsin. Chemical cross-linking and sedimentation velocity analysis showed that the lipase dimerizes due to removal of a region near its C-terminus. The lipase monomer has a sedimentation coefficient s20.w = 2.83 S, whereas the dimer has s20.w = 3.65 +/ 0.22 S. Hydrodynamic analysis using these sedimentation values and the masses determined by mass spectrometry indicated that the monomers are aligned side-by side in the dimer. An important change occurs in the apparent partial specific volume of the molecule upon activation. PMID- 8224198 TI - Cloning, expression and sequence analysis of cDNA for the Ca(2+)-binding photoprotein, mitrocomin. AB - The primary structure of mitrocomin consists of 190 amino acid residues, with three Ca(2+)-binding sites and a tyrosine residue at the C-terminus. Mitrocomin shows an amino acid sequence homology of 67.9% and 60.7% when compared with aequorin and clytin, respectively. The amino acid residues Cys152, His58, His169, Trp12, Trp86, Trp108, Trp129 and Trp173 are conserved in all three photoproteins, suggesting that they play a role in light emission. PMID- 8224199 TI - Receptor-like kinase activity in membranes of Arabidopsis thaliana. AB - A class of protein kinases in the plant Arabidopsis thaliana has been identified which has biochemical characteristics similar to those of the animal receptor protein kinases. These plant protein kinases are membrane-associated, glycosylated, exhibit a preference for Mn2+ relative to Mg2+, and range from 115 to 135 kDa when identified by renaturation after protein blotting. Evidence is presented that the TMK1 protein, a receptor-like protein kinase identified in Arabidopsis from its cloned gene, is also glycosylated in its native form, and shows a preference for Mn2+ over Mg2+ when assayed for kinase activity. All of the plant receptor-like protein kinases identified phosphorylate only serine and threonine residues. PMID- 8224200 TI - Delta opioid receptor mediates phospholipase C activation via Gi in Xenopus oocytes. AB - Cloned mouse delta-subtype opioid receptor (DOR1) was expressed in Xenopus oocytes to study the signal transduction. Opioid delta-agonists evoked a calcium dependent chloride current in oocytes injected with mRNA derived from DOR1, together with that from the alpha subunit of Gi1. The delta-agonist-induced current was blocked by naltrindol, a delta-specific antagonist. The delta-agonist evoked no or very weak currents in oocytes with the alpha subunit of Gq or G(o). These findings indicate the functional coupling between the opioid delta-receptor and phospholipase C through an activation of Gi. PMID- 8224201 TI - New isoforms of multifunctional calcium/calmodulin-dependent protein kinase II. AB - Calcium/calmodulin dependent protein kinase II (CaM kinase II) seems to act as an important regulator of intracellular signal transmission. Four subtypes, termed alpha to delta, have been cloned; some of them can exist as different splicing variants. All these isoforms share a great overall homology, and they contain 3 areas of low homology. We have identified 5 new variants of subtype delta so that the total number of different isoforms now adds up to 12. These variants are probably a result of different splicing and show several deletions in regard to subtype delta. The deletion sites do exactly match regions of low homology between the subtypes. This suggests a functional division of the CaM kinase II molecule into homologous and variable domains. The homologous domains are highly conserved. Therefore, it might be the case that the constitution of the variable domains is more significant for a certain isoform than its belonging to one of the 4 subtypes alpha to delta. PMID- 8224202 TI - Calcium-mobilizing effectors inhibit P-enolpyruvate carboxykinase gene expression in cultured rat hepatocytes. AB - Incubation of primary cultures of hepatocytes from fed and fasted rats with calcium ionophore strongly decreased glucose production from pyruvate. Like insulin, calcium ionophore A23187, phenylephrine, vasopressin, and prostaglandins E2 and F2 alpha caused a significant reduction (50-60%) in basal concentrations of mRNA for P-enolpyruvate carboxykinase (PEPCK), the main regulatory enzyme of gluconeogenesis. Phenylephrine, prostaglandin E2 and calcium ionophore A23187 were also able to counteract the induction of PEPCK gene expression by Bt2cAMP. These effects were similar to those exerted by both vanadate and phorbol ester TPA. The decrease in extracellular calcium by the addition of the calcium chelating agent EGTA to the incubation medium caused an increase in PEPCK mRNA levels. This effect was additive to that of Bt2cAMP and was counteracted by vanadate. PMID- 8224203 TI - Lysophosphatidylcholine mediates the mode of insertion of the NH2-terminal SIV fusion peptide into the lipid bilayer. AB - We report here on the interaction of a synthetic 12 residue peptide corresponding to the N-terminal sequence of gp32 from SIV with phospholipid bilayers. This peptide has been shown to induce lipid mixing of PC/PE/SM/Chol LUV (large unilamellar vesicles) at pH 7.4 and 37 degrees C [(1992) in: Advances in Membrane Fluidity, vol. 6, pp. 365-376, Wiley-Liss]. In the present study, this fusion process was inhibited by the addition of lysophosphatidylcholine (lysoPC) to the lipid bilayer of PC/PE/SM/Chol LUV. Fourier transform infrared spectroscopy (FTIR) reveals that the orientation of the SIV fusion peptide with respect to the lipid acyl chains depends on the presence of lysoPC in the lipid bilayer but that the peptide secondary structure and the amount of lipid-associated peptides do not depend on the lipid composition. The peptide is obliquely inserted into the lipid bilayer of vesicles without lysoPC, whereas it is oriented parallel to the lipid-water interface in the vesicles containing lysoPC. The data provide evidence that the orientation of the SIV fusion peptide depends on the lipid composition, and that this mediates its fusogenic activity. PMID- 8224204 TI - Formation of disulphide bonds in the reaction of SH group-containing amino acids with trimethylamine N-oxide. A regulatory mechanism in proteins. AB - Two amino acids containing SH group (cysteine and homocysteine)+trimethylamine N oxide systems were studied by FTIR and 1H NMR spectroscopy. This study demonstrates that cysteine and homocysteine ethylesters react with trimethylamine N-oxide. Immediately after mixing, SH...ON<==>S-...H+ ON hydrogen bonds with large proton polarizability are are formed. Then a reaction proceeds resulting in the formation of corresponding disulphides. Trimethylamine N-oxide is present in biological systems. Thus, our results suggest that trimethylamine N-oxide may play a regulatory role in S-S bond formation in enzymes and other proteins. PMID- 8224205 TI - Cyclosporine A protects mitochondria in an in vitro model of hypoxia/reperfusion injury. AB - Hypoxia/reperfusion injury is a major clinical problem. One of its hallmarks is an increased cytosolic Ca2+ content and an increased generation of reactive oxygen species in the cytosol and in mitochondria. In the present study of an in vitro model of hypoxia/reperfusion injury, mitochondria are exposed to Ca2+ in combination with extra- and intramitochondrially acting prooxidants. In this model mitochondria are damaged in a Ca(2+)-dependent manner. The extent and the site(s) of damage depend on both the kind of respiratory substrate and prooxidant used. The major damage occurs specifically at site I of the respiratory chain, and is due to hydrolysis of oxidized pyridine nucleotides and Ca2+ release followed by Ca2+ re-uptake (Ca2+ 'cycling'). Cyclosporine A completely protects against this damage. The protection is due to inhibition of pyridine nucleotide hydrolysis, an obligatory step in the sequence of events that links prooxidants to Ca2+ release from intact mitochondria. PMID- 8224206 TI - Interleukin-1 beta-induced cytosolic phospholipase A2 activity and protein synthesis is blocked by dexamethasone in rat mesangial cells. AB - Interleukin-1 beta induces gene expression and secretion of the secretory phospholipase A2 (sPLA2) and prostaglandin E2 (PGE2) release from rat mesangial cells. We have previously shown that prolonged treatment of rat mesangial cells with interleukin-1 beta (IL-1 beta) also enhances the cytosolic phospholipase A2 (cPLA2) activity. This effect of IL-1 beta on the cPLA2 activity is inhibited by actinomycin D and cycloheximide, indicating that both transcription and translation are involved. Here, we describe that IL-1 beta increases mRNA levels and protein synthesis of cPLA2 itself. In parallel with the effect of dexamethasone on the sPLA2, this glucocorticoid inhibits the IL-1 beta-enhanced cPLA2 activity as a result of the suppression of IL-1 beta-induced cPLA2 gene expression. This report suggests that the pro-inflammatory action of interleukin 1 beta may, in part, be mediated by its effects on cPLA2 activity. PMID- 8224207 TI - A comment on the absence of calcium regulation of human thioredoxin reductase. AB - It has been previously suggested that human thioredoxin reductase activity is regulated by calcium. However, the activity of a purified form of human placental thioredoxin reductase was found to not be affected by mM concentrations of calcium, well above intra- and extracellular physiological levels. Furthermore, the suggestion that an E-F hand is present in Escherichia coli thioredoxin reductase is strongly contested. These current results suggest that human thioredoxin reductase is not regulated by calcium. PMID- 8224208 TI - Chloroplast thylakoid protein phosphatase reactions are redox-independent and kinetically heterogeneous. AB - At least eleven thylakoid proteins become phosphorylated under reducing conditions, and redox titration has identified a common midpoint potential of Em = +38 +/- 4 mV, n = 0.95 +/- 0.06. In the presence of the phosphatase inhibitor NaF (10 mM), the redox dependency of phosphorylation is found to be essentially unchanged: Em = +50 +/- 3 mV, n = 1.02 +/- 0.04. Thylakoid membranes were phosphorylated in the light and then incubated at various redox potentials for 15 min in the dark; no redox dependency was observed in the dephosphorylation of any of the 17 bands then distinguishable by autoradiography and phosphorimaging. The phosphoprotein phosphatase reactions can be divided arbitrarily into four kinetic classes: the fastest, class I, includes LHC II; the moderate class II includes D1 and D2; the slow class III includes CP43 and the 9 kDa phosphoprotein; finally, a 19.5 kDa protein exhibited no loss of 32P at all. In separate experiments we measured thylakoid protein dephosphorylation initiated by changing the redox potential from -140 to +200 mV, in the presence or absence of fluoride. In this case the results are consistent with at least two kinetically distinguishable classes of phosphoprotein phosphatase reactions. We conclude that thylakoid protein phosphatase reactions are kinetically heterogeneous and redox independent. It follows that the redox dependency of thylakoid protein phosphorylation is a property of thylakoid protein kinase reactions. Our observed Em and n values are consistent with a primary site of kinase redox control at the level of PQ/PQ(.)- of the Qi (Qn) site of the cytochrome b6/f complex. PMID- 8224209 TI - No enzymatic activities are necessary for the stabilization of ascorbic acid by K 562 cells. AB - We disprove that living cells stabilize ascorbate by the activity of a trans plasma membrane semidehydroascorbate reductase. The two processes show different specificities for both substrate and inhibitor. Not only cells but also cell conditioned buffers stabilize ascorbate as long as compounds with molecular weights above 10 kDa are not removed. The effect is most probably due to chelation of traces of transition metals. PMID- 8224210 TI - K(+)-driven sinusoidal efflux of glutathione disulfide under oxidative stress in the perfused rat liver. AB - Tert-butyl hydroperoxide (BHP), hydrogen peroxide and diamide caused a rapid and simultaneous release of glutathione disulfide (GSSG) and K+ in the isolated perfused rat liver. Both BHP-induced effluxes were suppressed by prior depletion of hepatic glutathione, but not by co-infusion of desferrioxamine which prevented lipid peroxidation and cell death. High K+ media decreased the GSSG efflux even though hepatic GSSG levels remained high. The GSSG and K+ effluxes were repeatable if cellular K+ recovered after a short BHP exposure. Ouabain inhibited the K+ re-uptake and decreased the response to repeated BHP challenge. Thus, sinusoidal efflux of GSSG under oxidative stress may be driven by a K+ gradient. PMID- 8224211 TI - Interleukin-10 inhibits endotoxin-induced tissue factor mRNA production by human monocytes. AB - In Gram-negative septic shock, human monocytes synthesize and express on their cytoplasmic membrane tissue factor (TF), a potent activator of the coagulation cascades. The role of TF in triggering disseminated intravascular coagulation (DIC) in these patients appears to be clear. We report the suppressive effect of interleukin-10 (IL-10) on endotoxin-induced TF activity and antigen levels, and on the expression of TF mRNA levels in human monocytes. These results emphasize the potential therapeutic value of this cytokine in septic shock, a condition still associated with a high mortality rate. PMID- 8224212 TI - Methyl alpha-glycoside of N-thioacetyl-D-neuraminic acid: a potential inhibitor of influenza A virus. A 1H NMR study. AB - The binding of influenza A virus hemagglutinin to its cell surface receptor, alpha-linked 5-N-acetylneuraminic acid (sialic acid), was studied in solution. The effect of structural modifications introduced into the N-acetyl group of the sialic acid on the binding was monitored by determining the dissociation constants by proton nuclear magnetic resonance (1H NMR) spectroscopy. Methyl alpha-glycoside of N-thioacetylneuraminic acid showed high, whereas the corresponding N-methylcarbamoylneuraminic acid exhibited relatively low binding affinity towards the hemagglutinin. PMID- 8224213 TI - Effect of hyperthermia on the extracellular matrix. I. Heat enhances hyaluronan and inhibits sulphated glycosaminoglycan synthesis. AB - The synthesis of sulphated glycosaminoglycans (GAG) and hyaluronan (HA) was studied in 3 cell strains incubated at 37 degrees C or 42 degrees C. Cells were labelled with [3H]glucosamine and [35S]sulphate. No incorporation of [35S]sulphate was observed at 42 degrees C. Cellulose acetate electrophoresis of GAGs synthesized at 42 degrees C showed only one [3H]glucosamine-labelled band completely digested with Streptomyces hyaluronidase. The quantification of [3H]glucosamine-labelled GAGs before and after hyaluronidase treatment indicated an increased rate of HA synthesis at 42 degrees C as compared to the rate of synthesis at 37 degrees C. These results suggest that hyperthermia stimulates HA synthesis and inhibits sulphated GAG synthesis in several cell strains which in turn may contribute to the modification of extracellular environment in inflammation. PMID- 8224214 TI - Anion-mediated lysine-arginine interaction. Evidence in Chaetopterus variopedatus sperm protamine. AB - Chaetopterus ariopedatus sperm protamine is a stable oligomer. Specific amino acid side chain modifications show that the oligomeric structure depends on anion mediated lysine-arginine interactions. The occurrence of this type of interaction is confirmed by the finding that poly-L-arginine readily forms aggregates with poly-L-lysine or with the native but not with the protamine with carbamylated epsilon-amino groups. PMID- 8224215 TI - Watermelon glyoxysomal malate dehydrogenase is sorted to peroxisomes of the methylotrophic yeast, Hansenula polymorpha. AB - We have studied the fate of the watermelon (Citrullus vulgaris Schrad.) glyoxysomal enzyme, malate dehydrogenase (gMDH), after synthesis in the methylotrophic yeast, Hansenula polymorpha. The gene encoding the precursor form of gMDH (pre-gMDH) was cloned in an H. polymorpha expression vector downstream of the inducible H. polymorpha alcohol oxidase promoter. During methylotrophic growth, pre-gMDH was synthesized and imported into peroxisomes, where it was enzymatically active. The apparent molecular mass of the protein located in H. polymorpha peroxisomes was equal to that of pre-gMDH (41 kDa), indicating that N terminal processing of the transit peptide had not occurred in the yeast. PMID- 8224216 TI - Identification and characterisation of a type-1 protein phosphatase from the okadaic acid-producing marine dinoflagellate Prorocentrum lima. AB - The unicellular marine dinoflagellate, Prorocentrum lima, an established producer of okadaic acid (OA), was shown to contain a type-1 protein phosphatase (PP-1) the biochemical profile of which on Mono-Q and Superdex-75 fast protein liquid chromatography was identical to the catalytic subunit of PP-1 from rabbit skeletal muscle. Purified P. lima PP-1 (apparent molecular mass 37.5 kDa) was highly sensitive to inhibition by mammalian protein phosphatase inhibitor-1 and inhibitor-2, and to OA itself. A 6-7-fold increase in OA production by P. lima, when grown under controlled conditions, correlated with an up to 300-fold increase in P. lima PP-1 activity. Furthermore, P. lima did not contain any detectable type-2A protein phosphatase activity. This study represents the first identification of a serine/threonine protein phosphatase in a dinoflagellate. PMID- 8224217 TI - Molecular cloning of crustacean putative molt-inhibiting hormone (MIH) precursor. AB - A cDNA encoding the complete precursor of the putative molt-inhibiting hormone (MIH) of the shore crab, Carcinus maenas, was isolated and sequenced. The precursor consists of a putative 35 amino acid signal peptide and the 78 amino acid mature MIH. The deduced MIH amino acid sequence is in complete agreement with the sequence previously determined by Edman degradation. In situ hybridization revealed MIH-expression in a subpopulation of large neurosecretory perikarya of the medulla terminalis X-organ in the eyestalk. PMID- 8224218 TI - GTP-binding protein-activator sequences in the insulin receptor. AB - Some functions of the insulin receptor (insR) are assumed to be mediated by pertussis toxin-sensitive Gi/G(o) proteins. Here we have located G-protein activator domains in the cytoplasmic region of the human insR. We searched the sequence of insR and found three candidate regions at residues 1039-1061, 1147 1168 and 1325-1345, referred to as ISRP1, ISRP2 and ISRP3, respectively. Among them, the Gi/G(o)-activating function was observed only in peptide ISRP3. ISRP1 specifically activated Gs, whereas ISRP2 had no effect on G proteins. ISRP2 and ISRP3 contained five of six autophosphorylated tyrosine residues in insR. After tyrosine phosphorylation, ISRP2 showed specific Gi-activating function, and ISRP3 potentiated its ability and became capable of activating G proteins generally. This is the first study that specifies G-protein-activator domains in insR and describes their modification by autophosphorylation. PMID- 8224219 TI - Identification of casein kinase II as a major endogeneous caldesmon kinase in sheep aorta smooth muscle. AB - A caldesmon kinase activity was detected in an ATP extract of the myofibril-like pellet from sheep aorta. The enzyme was purified 745-fold and was identified as casein kinase II on the basis of molecular size, substrate specificity, and high sensitivity to heparin inhibition. Casein kinase II phosphorylated isolated caldesmon and caldesmon incorporated into native thin filaments, and transferred about 1 mol of phosphate per mol of caldesmon-h. Ser-73 was the main site phosphorylated by casein kinase II in chicken gizzard caldesmon. Phosphorylation of caldesmon reduced its affinity for smooth muscle myosin but had no effect upon the ability of caldesmon to inhibit the ATPase activity of actomyosin. PMID- 8224221 TI - The VIP2 receptor: molecular characterisation of a cDNA encoding a novel receptor for vasoactive intestinal peptide. AB - We have cloned and sequenced a cDNA (RPR4) encoding a new member of the secretin/calcitonin/parathyroid hormone (PTH) receptor family. RPR4 was identified by PCR of rat pituitary cDNA, and a full-length clone was isolated from a rat olfactory bulb cDNA library. When RPR4 was functionally expressed in COS 7 cells, cyclic adenosine monophosphate (cAMP) production was stimulated by vasoactive intestinal peptide (VIP), pituitary adenylate cyclase activating peptides (PACAP-38 and PACAP-27) and helodermin, with equal potency. Peptide histidine isoleucine (PHI) and rat growth hormone releasing hormone (rGHRH) also stimulated cAMP production at lower potency. This suggests that RPR4 encodes a novel VIP receptor which we have designated the VIP2 receptor. In situ hybridisation showed that mRNA for this receptor was present mainly in the thalamus, hippocampus and in the suprachiasmatic nucleus. PMID- 8224220 TI - Molecular characterization of a blue visual pigment gene in the fish Astyanax fasciatus. AB - We report here the isolation and sequence determination of a gene closely linked to the Astyanax red visual pigment gene. Reverse transcription polymerase chain reaction assays show that this new gene (B23 Af) and the previously characterized red and green visual pigment genes of Astyanax are all expressed in the eye. Phylogenetic analysis shows that B23Af belongs to the group consisting of short wavelength-sensitive pigment genes from different species and is most closely related to the goldfish blue visual pigment gene. PMID- 8224222 TI - ADP-ribosylation of Rho proteins inhibits sperm motility. AB - The highly homologous Rho proteins RhoA, RhoB and RhoC are low-molecular-mass GTP binding proteins. They are selectively ADP-ribosylated by Clostridium botulinum ADP-ribosyltransferase C3 (C3 exoenzyme). The biological function of the Rho proteins is still unclear; there is evidence that they are involved in the regulation of the filamental network of cells. Here we report that C3 exoenzyme like toxins ADP-ribosylate small GTP-binding proteins in bovine spermatozoa and inhibit sperm motility. These findings indicate that Rho proteins which reportedly regulate the microfilament system are basically involved in sperm motility. PMID- 8224223 TI - Characterization of AWN-1 glycosylated isoforms helps define the zona pellucida and serine proteinase inhibitor-binding region on boar spermadhesins. AB - Spermadhesin AWN-1 (14 kDa) belongs to a recently described family of boar sperm surface-associated proteins. AWN-1 is a multifunctional protein which possesses heparin-, serine proteinase inhibitor-, and zona pellucida glycoprotein-binding capability. Therefore it has been implicated in sperm capacitation and sperm oocyte attachment. Here, we report the characterization of 22-25 kDa isoforms of AWN-1 isolated by heparin-affinity chromatography, which fail to bind to zona pellucida glycoproteins or serine proteinase inhibitors. Our results show that the structure of the high and low molecular mass AWN-1 forms differ in that the former is N-glycosylated at Asp50 and truncated at the C-terminus. The inability of the glycosylated AWN-1 molecules to bind ligands is due solely to the presence of the oligosaccharide moieties, however. This indicates that glycosylation of AWN-1 may modulate its ligand-binding capabilities. On the other hand, the effect of glycosylation on ligand-binding suggests that both the zona pellucida- and the serine proteinase inhibitor binding domain(s) may be located around the glycosylation point. PMID- 8224224 TI - Attraction of human monocytes by the neuropeptide secretoneurin. AB - Secretoneurin is a newly discovered 33-amino-acid peptide derived from secretogranin II (chromogranin C) that is found in sensory afferent C-fibers. We show here that secretoneurin triggers the selective migration of human monocytes in vitro and in vivo. Combinations of secretoneurin with the sensory neuropeptides, substance P or somatostatin, synergistically stimulate such migration. The attraction of monocytes represents the first established function of secretoneurin as a sensory neuropeptide. PMID- 8224225 TI - The sarco(endo)plasmic reticulum Ca(2+)-ATPase mRNA isoform, SERCA 3, is expressed in endothelial and epithelial cells in various organs. AB - The sarco(endo)plasmic reticulum Ca(2+)-ATPase mRNA isoform, SERCA 3, was previously shown to be expressed in a great variety of muscle and non-muscle tissues [(1989) J. Biol. Chem. 264, 18568] but its cellular localization within these organs was unknown. We have used in situ hybridization and RNase protection techniques to demonstrate that SERCA 3 mRNA is expressed in specific cell types, namely the endothelial and epithelial cells. PMID- 8224226 TI - Studies on the ability of minor groove binders to induce supercoiling in DNA. AB - The effect of various non-intercalating minor groove binders on closed circular DNA in the presence of topoisomerase I has been studied by means of agarose gel electrophoresis. Analogues of the netropsin series (lexitropsins) and SN-6999 effectively produce positive supercoils, as indicated by analysis of the topoisomers in the presence of chloroquine and the evaluated linking number changes. Analogues of the distamycin series are less effective, and bisquaternary heterocycles, as well as DAPI and pentamidine, were found to be ineffective ligands. The large differences observed in the ability of minor groove binders to induce positive supercoils are discussed. PMID- 8224227 TI - Expression of fully active ammodytoxin A, a potent presynaptically neurotoxic phospholipase A2, in Escherichia coli. AB - A cDNA encoding the most presynaptically neurotoxic phospholipase A2, ammodytoxin A, from the venom of the long-nosed viper (Vipera ammodytes ammodytes) has been expressed in Escherichia coli. Ammodytoxin A was produced as a fusion protein with the 81 N-terminal residues of adenylate kinase followed by the tetrapeptide recognition site for factor Xa (IEGR) just preceding the first amino acid residue of the toxin. The fusion protein was expressed under the control of tac promoter without IPTG induction in the form of insoluble inclusion bodies. It was dissolved in guanidine hydrochloride, S-sulfonated and refolded in a reoxidation mixture including a reduced/oxidized glutathione redox couple. Ammodytoxin A was fully activated by limited hydrolysis with trypsin that preferentially cleaves the fusion protein at the factor Xa recognition site and purified by cation exchange chromatography. The correct N-terminus was confirmed by protein sequencing. Recombinant ammodytoxin A has been proved to be indistinguishable from the native toxin in its enzymatic activity and toxicity. PMID- 8224228 TI - Purification and characterization of p27, a protein from hepatocyte chromatin. Evidence suggesting that it binds selectively to guanine-rich single-stranded DNA. AB - A protein designated p27 that binds preferentially to single-stranded DNA rich in guanine tracts was purified to near homogeneity from rabbit hepatocyte non histone protein extract. Purified p27 migrated as a 27 kDa polypeptide on denaturing SDS-PAGE and displayed a native molecular mass of approximately 155 kDa on Sephadex G-150 or Sepharose 6B-Cl gel filtration columns. Gel shift analysis indicated that maximum binding of p27 to single-stranded DNA required the presence of tracts of four or more contiguous guanine residues. The lowest found dissociation constant, 1.4 x 10(-8) M/l, was for single-stranded DNA that contained a (dG)17 run. PMID- 8224229 TI - Increased expression of type VI collagen genes in drug-induced gingival enlargement. AB - Fibrotic gingival enlargements induced by phenytoin or nifedipine were examined with special reference to type VI collagen expression. Immunolocalization studies showed abnormal accumulation of type VI collagen around the collagen fiber bundles in the fibrotic gingival enlargements. Examination of total RNA extracted from fibroblasts and tissues of enlarged gingivae demonstrated increased type VI collagen steady-state mRNA levels. These results suggest that excessive deposition of type VI collagen in drug-induced gingival enlargement is attributed to increased expression of the collagen genes. PMID- 8224230 TI - Expression of non-hepatic-type S-adenosylmethionine synthetase isozyme in rat hepatomas induced by 3'-methyl-4-dimethylaminoazobenzene. AB - It is known that a high incidence of hepatocellular carcinoma in rat liver can be induced by such azo dye carcinogens as 3'-methyl-4-dimethylaminoazobenzene (3' MeDAB). Mammalian S-adenosylmethionine (AdoMet) synthetase exists as two isozymes, non-hepatic-type and liver-type enzymes, which are the products of two different genes. We have examined the expression of two AdoMet synthetase isozyme proteins and mRNAs in rat hepatomas induced by 3'-Me-DAB. The levels of non hepatic-type enzyme protein and mRNA are clearly induced by 3'-Me-DAB feeding. On the other hand, the levels of liver-type enzyme protein and mRNA are nearly the same or slightly decreased during hepatocarcinogenesis. These results indicate that the expression of the non-hepatic-type isozyme gene is obviously influenced with the progression of carcinogenesis and that the non-hepatic-type isozyme is useful as a oncodevelopmental marker in the liver. PMID- 8224231 TI - Enzymatic oxidation of the bifunctional wheat inhibitor of subtilisin and endogenous alpha-amylase. AB - Oxidation of the bifunctional wheat inhibitor of subtilisin and endogenous alpha amylase catalyzed by horseradish peroxidase results in the loss of the inhibitory activity against both enzymes. The enzymatic oxidation is accompanied by modification of one methionine and two tryptophan residues in the protein. The results obtained, together with data on chemical modification and limited proteolysis, allow us to conclude that Met34-Ala35 is the reactive site of the inhibitor responsible for the interaction with subtilisin. It is supposed that the reactive site of the inhibitor responsible for the interaction with alpha amylase contains one or two tryptophan residues. PMID- 8224232 TI - Purification of a vasoactive peptide related to lysyl-bradykinin from trout plasma. AB - Incubation of plasma from the steelhead trout, Oncorhynchus mykiss with porcine pancreatic glandular kallikrein generated bradykinin-like immunoreactivity. The primary structure of the immunoreactive peptide was established as: Lys-Arg-Pro Pro-Gly-Trp-Ser-Pro-Leu-Arg. This sequence shows two amino acid substitutions (Phe6-->Trp and Phe9-->Leu) compared with mammalian lysyl-bradykinin (kallidin). Bolus intraarterial injection of the purified peptide produced a strong and sustained vasopressor response in the unanaesthetized trout. The data demonstrate that the kallikrein-kinin system predates the appearance of tetrapods and suggest a role for this system in cardiovascular regulation in fish. PMID- 8224233 TI - Trimeric forms of the photosystem I reaction center complex pre-exist in the membranes of the cyanobacterium Spirulina platensis. AB - Oligomeric and monomeric forms of chlorophyll-protein complexes of photosystem I (PSI) have been isolated from the mesophilic cyanobacterium Spirulina [(1992) FEBS Lett. 309, 340-342]. Electron microscopic analysis of the complexes showed that the oligomeric form is a trimer of the shape and dimensions similar to those of the trimer from thermophilic cyanobacteria. The chlorophyl ratio in the isolated trimer and monomer was found to be 7:3. The trimeric form of PSI complex in contrast to the monomeric one contains the chlorophyll emitting at 760 nm (77K), which is also found in Spirulina membranes and therefore could be used as an intrinsic probe for the trimeric complex. The 77K circular dichroism spectrum of the trimeric form is much more similar to that of Spirulina membranes than the spectrum of the monomer. Thus, the trimeric PSI complexes exist and dominate in the Spirulina membranes. PMID- 8224234 TI - Effects of repetitive stimulation, veratridine and ouabain on cytoplasmic pH in frog nerve fibres: role of internal Na+. AB - Changes of cytoplasmic pH (pHi) in frog nerve fibres during repetitive stimulation have been measured using the fluorescent pH indicator dye fluorescein diacetate (FDA). Under control conditions repetitive (10-50 Hz) stimulation caused only a very small decrease in pHi (by 0.015-0.06 pH units). Modification of Na+ channels by veratridine (VER, 10 microM) greatly increased this stimulus evoked (SE) internal acidification. Blockade of the Na(+)-K+ pump by ouabain (0.5 mM) enhanced the effects VER and prevented pHi recovery after the termination of repetitive stimulation. A similar inhibition of post-stimulatory recovery of pHi was observed after replacement of external Na+ with Li+, which is not accepted by the Na(+)-K+ pump instead of Na+. These data suggest that SE intracellular acidification in nerves results from or is closely associated with an increase in [Na+]i. Treatments that promote Na+ influx and accumulation of Na+ inside the fibre enhance reduction of pHi. Li+ can be substituted for Na+ in this process. PMID- 8224235 TI - Site-directed mutagenesis of AMP-binding residues in adenylate kinase. Alteration of substrate specificity. AB - Adenylate kinase is highly specific for AMP as phosphoryl acceptor. We have found that the replacement of Thr39 by Ala in the chicken muscle enzyme, alone or together with the replacement of Leu66 by Ile, caused remarkable increases in CMP and UMP activities with a concomitant decrease in AMP activity; therefore, the resulting mutant enzymes show CMP and UMP activities/AMP activity ratios much higher than the wild-type enzyme. The mutant enzyme in which Ala is substituted for Thr39 has a Vmax value for CMP comparable to that of CMP-UMP kinase. PMID- 8224236 TI - Combined effects of diffusional hindrances, electrostatic repulsion and product inhibition on the kinetic properties of a bound acid phosphatase. AB - The kinetic properties of a soybean cell wall phosphatase were studied and compared under different environmental conditions. The native enzyme isolated from the wall exhibits classical Michaelis-Menten kinetics. When buried into the cell wall, its natural environment, the enzyme mimics an apparent negative cooperativity. This deviation from hyperbolic dependence of the activity vs. the substrate concentration may result from an heterogeneous distribution of the enzyme molecules between the surface and the inside of the wall, where an electrostatic partition effect takes place. Cell wall fragments compacted on a Millipore-type membrane allow the study of the dual effects of diffusional hindrances and electrostatic interactions on the global kinetic behavior. By increasing the ionic strength, partition effects can be suppressed and therefore diffusional effects alone can be taken into account. On the one hand, diffusion and partition act individually and synergetically to decrease the apparent global affinity of phosphatase with respect to glucose 6-phosphate. On the other hand, product inhibition by inorganic phosphate is subjected to dual effects from diffusion through accumulation and partition through repulsion. A simple diffusion-partition reaction model accounts qualitatively and quantitatively for the experimental observations. PMID- 8224237 TI - Regulation of adenylyl cyclase from Blastocladiella emersonii by guanine nucleotides. AB - GTP gamma S stimulates adenylyl cyclase in particulate fractions of Blastocladiella emersonii zoospores. Cholera toxin catalyses the ADP-ribosylation of a membrane protein of a molecular weight (46,000) similar to that of the alpha subunit of Gs found in vertebrate cells. A membrane protein of 46 kDa can also be recognized in Western blots by an antipeptide antiserum (RM/1) raised against the C-terminus of G alpha 2-subunits. These results suggest that a G-protein mediates the regulation of Blastocladiella adenylyl cyclase by guanine nucleotides. PMID- 8224238 TI - Mosses do express conventional, distantly B-type-related phytochromes. Phytochrome of Physcomitrella patens (Hedw.). AB - We have screened a cDNA library of the moss Physcomitrella patens (Hedw.) for phytochrome sequences. The isolated sequences turned out to encode a phytochrome dissimilar to the phytochrome type postulated for the moss Ceratodon [(1992) Plant Mol. Biol. 20, 1003-1017] Physcomitrella phytochrome was completely alignable to fern phytochrome (Selaginella) and phytochromes of higher plants. The frequency of clones encoding this phytochrome indicated that a Ceratodon-like type should only be expressed, if at all, with lower frequencies than the sequenced phytochrome cDNA. Sequence differences between lower plant phytochromes are small as compared to phytochrome types of higher plants. PMID- 8224239 TI - Molecular cloning of cDNA for the 29 kDa proteinase participating in decomposition of the larval fat body during metamorphosis of Sarcophaga peregrina (flesh fly). AB - A cDNA clone for the 29 kDa proteinase participating in tissue disintegration during metamorphosis of Sarcophaga was isolated. This proteinase, named Sarcophaga cathepsin B, consisted of 256 amino acid residues, and contained three putative N-glycosylation sites. By comparison with other cathepsins B, its unique substrate specificity was partly explained by Ala at position 248. PMID- 8224240 TI - Na+,K(+)-ATPase of gastric cells. A target of Helicobacter pylori cytotoxic activity. AB - The present study shows a direct impairing action of a cytotoxin-producing Helicobacter pylori strain on the Na+,K(+)-ATPase (evaluated as K(+)-dependent phosphatase activity) of human gastric epithelial cells in culture. The toxin itself is likely involved in this action which may also account for the cell edema found in vivo in Helicobacter pylori-colonized stomach. PMID- 8224241 TI - Inhibition of the epidermal growth factor receptor tyrosine kinase activity by leflunomide. AB - The active metabolite of leflunomide, A77 1726 inhibits the proliferation of a variety of mammalian cell lines in culture. Epidermal growth factor (EGF) dependent proliferation is inhibited by A77 1726 at an effective dose of 30-40 microM. A77 1726 appears to directly inhibit the EGF receptor tyrosine-specific kinase activity both in intact cells and purified EGF receptors at the same effective dose. These data suggest that leflunomide inhibits cellular proliferation by the inhibition of tyrosine-specific kinase activities. PMID- 8224242 TI - The antirepressor of phage P1. Isolation and interaction with the C1 repressor of P1 and P7. AB - Two antirepressor proteins, Ant1 and Ant2, of molecular weight 42 and 32 kDa, respectively, are encoded by P1 as a single open reading frame, with the smaller protein initiating at an in-frame start codon. Another open reading frame, icd, 5' upstream of and overlapping ant1 is required for ant1 expression. Using appropriate ant gene-carrying plasmids we have overproduced and purified Ant1/2 in the form of a protein complex and Ant2 as a single protein. Sequence analysis confirmed the N-terminal amino acids predicted from the DNA sequence of ant1/ant2, except that the N-terminal methionine is missing in the Ant2 protein. Under appropriate conditions the C1 repressors of phages P1 and P7 specifically co-precipitate with the Ant1/2 complex but not with Ant2 protein alone. The results suggest that the antirepressor may exert its C1-inactivating function by a direct protein-protein interaction. PMID- 8224243 TI - Inhibition of low-density lipoprotein oxidation by nitric oxide. Potential role in atherogenesis. AB - The effects of nitric oxide (.NO) and nitrovasodilators on the oxidation of low density lipoprotein (LDL) have been studied. S-Nitroso-N-acetylpenicillamine (SNAP) and sodium nitroprusside (SNP) inhibited Cu(2+)- and 2,2'-azobis-2 amidinopropane hydrochloride-dependent oxidation of LDL as monitored by oxygen consumption and the formation of thiobarbituric acid-reactive substances, conjugated dienes, and lipid hydroperoxides. In the case of SNP, inhibition of LDL oxidation occurred only when the incubation mixture was irradiated with visible light. SNAP, however, exerted a dose-dependent inhibition of Cu(2+) catalyzed oxidation of LDL even in the dark. Addition of .NO dissolved in deoxygenated buffer also inhibited the progression of LDL oxidation. Mouse peritoneal macrophages were less able to degrade LDL that had been oxidized in the presence of SNAP. Using an .NO electrode, it was estimated that a continuous production of .NO (< or = 760 nM/min) could retard the progression of LDL oxidation. We propose that .NO can inhibit LDL oxidation by acting as a chain breaking antioxidant that is capable of scavenging carbon-centered and peroxyl radicals. Biological implications of this novel .NO antioxidant property are discussed in relation to atherogenesis and contrasted to the prooxidant property of .NO when generated in the presence of superoxide. PMID- 8224244 TI - Rab11, a small GTPase associated with both constitutive and regulated secretory pathways in PC12 cells. AB - A specific polyclonal antibody was used to investigate the subcellular distribution of the small GTPase, rab11p, in the neuroendocrine cell line, PC12. We took advantage of a previously described pulse-chase protocol based on sulfation to examine the distribution of rab11 along the secretory pathway. Using the rab11 antiserum, but not serum depleted of rab11 antibodies, we were able to specifically immunoisolate markers of the constitutive and the regulated secretory pathway in the trans-Golgi network (TGN) as well as after their exit from this compartment (constitutive secretory vesicles, immature, and mature secretory granules). We therefore conclude that rab11p is associated with the TGN and with TGN-derived vesicles of both the constitutive and the regulated secretory pathway in PC12 cells. PMID- 8224245 TI - Membrane phospholipid asymmetry in DMPC-induced human red cell vesicles. AB - Vesicles that do not contain spectrin were released from human erythrocytes by incubation with dimyristoylphosphatidylcholine. The transbilayer orientation of membrane phospholipids was subsequently determined by two independent methods. Incubation with phospholipase A2 revealed that the phospholipid asymmetry observed in red blood cells was essentially preserved in vesicles. By use of the prothrombinase assay a still highly asymmetric distribution of phosphatidylserine could be demonstrated in spite of its slightly increased exposure on the vesicle surface. These results show that membrane phospholipid asymmetry can be maintained in a system that does not contain an intact membrane skeleton or spectrin. PMID- 8224246 TI - Differentiation of HL60 promyelocytic cells is promoted by a 'differentiation enhancing factor' produced by erythroleukemia cells. AB - A differentiation enhancing factor isolated from murine erythroleukemia cells is also a potent enhancer of the differentiation of HL60 human promyelocytic leukemia cells, induced by retinoic acid and by phorbol ester. This stimulating effect is the result of a large increase in the sensitivity of HL60 cells for retinoic acid and for phorbol 12-myristate 13-acetate (20-fold and 40-fold, respectively). Accelerated differentiation induced by the protein factor, and monitored by the appearance of marker enzymes, is accompanied by a large increase in the fluctuation of the levels of protein kinase C (PKC) isozymes in HL60 cells. These results provide further support for the role of this new protein factor in cell differentiation and indicate that other cell types are susceptible to its biological effect. PMID- 8224247 TI - X-arrestin: a new retinal arrestin mapping to the X chromosome. AB - We have been using a differential cDNA cloning approach to isolate human retina specific and retina-enriched genes [1]. A 1,314 bp cDNA was isolated by this approach, representing a highly retina-specific message encoding a 388 amino acid protein showing 58%, 50%, and 49% homology to bovine beta-arrestin, and bovine and human retinal arrestin (S-antigen), respectively. Chromosomal mapping localized this new arrestin gene to the proximal long arm of the X chromosome, hence it was named X-arrestin. In situ hybridization demonstrated its expression in the inner and outer segments and the inner plexiform regions of the retina. PMID- 8224248 TI - In vitro characterization of Ro 46-2005, a novel synthetic non-peptide endothelin antagonist of ETA and ETB receptors. AB - Ro 46-2005 is a new synthetic non-peptide endothelin (ET) receptor antagonist. In binding experiments, Ro 46-2005 proved to be equipotent (IC50 200-500 nM) for inhibition of [125]ET-1 binding on the two known ET receptor subtypes (ETA and ETB). Scatchard analysis was consistent with a competitive binding mode. Ro 46 2005 also inhibited the functional consequences of ET-1 stimulation: the ET-1 induced release of arachidonic acid from rat mesangial cells was inhibited with an IC50 of 1.8 microM. PMID- 8224249 TI - Engineering resistance to 'aging' of phosphylated human acetylcholinesterase. Role of hydrogen bond network in the active center. AB - Recombinant human acetylcholinesterase (HuAChE) and selected mutants (E202Q, Y337A, E450A) were studied with respect to catalytic activity towards charged and noncharged substrates, phosphylation with organophosphorus (OP) inhibitors and subsequent aging of the OP-conjugates. Amino acid E450, unlike residues E202 and Y337, is not within interaction distance from the active center. Yet, the bimolecular rates of catalysis and phosphylation are 30-100 fold lower for both E450A and E202Q compared to Y337A or the wild type and in both mutants the resulting OP-conjugates show striking resistance to aging. It is proposed that a hydrogen bond network, that maintains the functional architecture of the active center, involving water molecules and residues E202 and E450, is responsible for the observed behaviour. PMID- 8224250 TI - Long-term effects of thrombin require sustained activation of the functional thrombin receptor. AB - Thrombin is a potent activator of human glomerular epithelial cells (HGEC). Here we compare short-term and long-term effects of thrombin and thrombin receptor agonist peptide (TRAP) which selectively activates the functional thrombin receptor. TRAP, as thrombin, increases intracellular free Ca2+ concentration and acts synergistically with growth factors possessing tyrosine kinase receptors on DNA synthesis. Thrombin induces synthesis of proteins of the fibrinolytic system and cell proliferation if it is present for at least 8 h. TRAP alone does not stimulate protein synthesis and is not mitogenic. However, in the presence of the aminopeptidase inhibitor amastatin all long-term effects of thrombin can be fully mimicked by TRAP. In conclusion, different effects of thrombin and TRAP may be related to the degradation of TRAP by cellular ectoenzymes. The recently cloned thrombin receptor accounts for early intracellular signals and long-term cellular effects that require sustained activation of this receptor. PMID- 8224251 TI - Expression of interleukin-1 alpha and interleukin-1 receptor type I genes in murine brown adipose tissue. AB - At thermoneutral conditions, high steady-state levels of transcripts for both IL 1 alpha and its receptor IL-1RtI were found in specialized thermogenic organ, brown adipose tissue (BAT) of adult mice, as compared with the levels in lymph nodes, brain and spleen. A pronounced decrease of IL-1 alpha mRNA level in BAT was observed after lipopolysaccharide (LPS) administration and after exposure to cold. Likewise, LPS decreased the IL-1RtI mRNA level and depressed also the expression of cold-inducible genes for the BAT-specific heat-producing uncoupling protein and for lipoprotein lipase. It is concluded that, besides the centrally mediated effects, there exists a direct peripheral interaction of IL-1 cytokines with BAT cells. PMID- 8224252 TI - Cloning and expression in yeast of a higher plant chorismate mutase. Molecular cloning, sequencing of the cDNA and characterization of the Arabidopsis thaliana enzyme expressed in yeast. AB - Chorismate mutase (EC 5.4.99.5) catalyzes the first step in the branch of the shikimate pathway which leads to the aromatic amino acids, phenylalanine and tyrosine. We have isolated a cDNA for this enzyme from the higher plant, Arabidopsis thaliana, by complementing a yeast strain (aro7) with a cDNA library from A. thaliana. This is the first chorismate mutase cDNA isolated from a plant. It encodes a protein of 334 amino acids. The identity of the deduced amino acid sequence is 41% to the chorismate mutase sequence from Saccharomyces cerevisiae. The N-terminal portion of the deduced amino acid sequence has no homology to the S. cerevisiae sequence but resembles known plastid-specific transit peptides. The A. thaliana chorismate mutase expressed in yeast revealed allosteric control by the three aromatic amino acids, as previously described for plastidic chorismate mutase isozymes. PMID- 8224253 TI - The secondary structure of the ferredoxin transit sequence is modulated by its interaction with negatively charged lipids. AB - Import of proteins into chloroplasts depends on an N-terminal transit sequence. Transit sequences contain little primary sequence similarity and therefore recognition of these sequences is thought to involve specific folding. To assess the conformational flexibility of the transit sequence, we studied the transit peptide of preferredoxin (trfd) by circular dichroism. In buffer, trfd is in a random coil conformation. A large increase in alpha-helix was induced in the presence of micelles or vesicles formed by anionic lipids. Less pronounced changes in secondary structure were induced by zwitterionic detergents but no changes were observed in the presence of neutral detergents or vesicles composed of phosphatidylcholine. PMID- 8224254 TI - Primary structure and catalytic properties of extracellular ribonuclease of Bacillus circulans. AB - A complete amino acid sequence of extracellular Bacillus circulans RNase was established and compared with a structure of B. amyloliquefaciens RNase. Gln15, Gly65 and Gln104 in B. amyloliquefaciens RNase were found to be replaced by Leu, Ala and Lys, respectively, in B. circulans RNase. Catalytic properties of B. circulans RNase were studied. PMID- 8224255 TI - Creative dialogue. PMID- 8224256 TI - On reporting pregnancies by assisted reproductive technology. PMID- 8224257 TI - Collection and report of assisted reproductive technology outcome data: problems and solutions. PMID- 8224258 TI - Adhesion formation after laparoscopic ovarian cautery for polycystic ovarian syndrome: lack of correlation with pregnancy rate. AB - OBJECTIVE: To assess adhesion formation after laparoscopic ovarian cautery in women with polycystic ovarian syndrome (PCOS) and the efficacy of Interceed Adhesion Barrier (Ethicon, Summerville, NJ) in their prevention. DESIGN: Prospective, randomized, blinded, clinical study of laparoscopic ovarian cautery with application of Interceed to one ovary, followed by short interval second look laparoscopy, scoring of adhesions, and clinical follow-up. SETTING: Tertiary care clinic at a University teaching hospital. PATIENTS: Eight infertile women with PCOS who failed to conceive with previous clomiphene citrate (CC) therapy. RESULTS: Periovarian adhesions of varying severity developed in all women after laparoscopic ovarian cautery. Interceed showed no protective effect. Despite this finding, all women initiated regular menses after laparoscopic ovarian cautery and seven of eight women spontaneously conceived eight singleton pregnancies without any further therapy. CONCLUSION: Laparoscopic ovarian cautery should be considered in infertile women with PCOS who fail to respond to CC therapy. These women must be counseled with respect to the possible complication of postoperative adhesion formation. PMID- 8224259 TI - Initial experience of a new linear everting falloposcopy system in comparison with hysterosalpingography. AB - OBJECTIVE: To assess the endolumenal portion of the fallopian tube from ostium to fimbria using a specially designed linear everting catheter and microendoscope. The study compared falloposcopy results with those obtained with hysterosalpingography (HSG) in the same patients and explored the system's ability to classify internal tubal conditions. DESIGN: A prospective study of a new diagnostic technique, falloposcopy, versus conventional means of evaluating tubal status. SETTING: A tertiary university infertility center. PATIENTS: Eighteen infertile patients with a previous history of infertility of at least 2 years duration. INTERVENTION: Diagnostic falloposcopy was performed on a total of 31 tubes. RESULTS: Although the results of HSG and falloposcopy were in agreement in 19 cases, falloposcopy findings were at significant variance with HSG in 12 cases (40%). Seventeen of the tubes visualized by falloposcopy were considered to be normal, 10 showed mild disease, and 4 were severely damaged. CONCLUSION: Falloposcopy using the linear everting catheter is rapid and atraumatic. It provides more complete information concerning tubal status than HSG and as such constitutes an important advance in diagnosis and therapy planning. PMID- 8224260 TI - The efficiency of transvaginal ultrasonography in the diagnosis of endometrioma. AB - OBJECTIVE: To assess the efficiency of transvaginal ultrasonography in the screening of endometriomas and in differentiating endometriomas from other ovarian masses. DESIGN: Prospective study with surgical and pathological confirmation of the sonographic diagnosis. SETTING: Department of Obstetrics and Gynecology at the University of Cagliari, Cagliari, Italy. PATIENTS: All premenopausal nonpregnant women (n = 236) submitted to laparotomy or laparoscopy from May 1991 to December 1992. INTERVENTIONS: Transvaginal ultrasonographic examinations were done within 1 week before surgery. Ultrasonographic diagnosis was compared with surgical and histopathological findings. MAIN OUTCOME MEASURE: The visualization of round-shaped homogeneous hypoechoic "tissue" of low-level echoes within the ovary was chosen as characteristic ultrasonographic finding of endometrioma. Sensitivity, specificity, and predicted values were calculated for each visualized ovary (group A) to assess the efficiency in the screening, and for each visualized cyst (group B) to assess the efficiency in differentiating endometriomas from other ovarian masses. RESULTS: Four hundred twenty-three of 466 ovaries were visualized. Of the 82 visualized cysts, 27 were suspected to be endometriomas, and the diagnosis was confirmed by pathology in 21. The sensitivity and the specificity were 75% and 99%, respectively, in group A, and 84% and 90%, respectively, in group B. CONCLUSION: The study demonstrates that transvaginal ultrasonography has an efficiency of 88% in differentiating endometriomas from other ovarian masses with a specificity of 90%. PMID- 8224261 TI - The inability of preovulatory ovarian scan to predict multifetal pregnancy occurrence in a follow-up of induction of ovulation with menotropins. AB - OBJECTIVE: To establish the predictive role of preovulatory ovarian ultrasonography in the occurrence of multiple pregnancy after hMG and hCG treatment for anovulatory infertility. DESIGN: Prospective. SETTING: Outpatient Infertility Clinic. PATIENTS: Ninety-five anovulatory women who conceived after gonadotropin therapy. INTERVENTION: Induction of ovulation by hMG and hCG monitored by plasma E2 measurements and ovarian ultrasonography. MAIN OUTCOME MEASURES: All follicles visualized on the day of hCG administration were recorded and divided into the following four groups: group I, 10 to 12 mm; group II, 13 to 15 mm; group III, 16 to 18 mm; and group IV, 19 mm and larger. The sonographic findings were statistically evaluated to 80 singletons and 45 multiple pregnancies. RESULTS: No statistical correlation was found to exist between the number of follicles from the different groups and the number of fetuses. CONCLUSIONS: The number and sizes of follicles visualized on the day of hCG administration have no predictive value regarding the occurrence of a multiple pregnancy. PMID- 8224262 TI - The value of luteal support with progesterone in gonadotropin-induced cycles. AB - OBJECTIVE: To evaluate the effect of luteal support with vaginal P suppositories in hMG- and hCG-induced cycles. DESIGN: Between March 1988 and February 1989 patients did not receive luteal support, while between March 1989 and January 1990 P was given routinely in the luteal phase. Induction protocol and patient selection remained otherwise unchanged. SETTING: Infertility clinic of a tertiary care hospital. PATIENTS: Twenty-seven patients with hypogonadotropic amenorrhea (World Health Organization [WHO] group I) (11 women with luteal support, 16 women without) and 102 patients with euprolactinemic clomiphene citrate (CC)-resistant anovulation (WHO group II) (52 women with luteal support, 50 women without). INTERVENTION: Vaginal P suppositories 200 mg/d in the luteal support group. MAIN OUTCOME MEASURES: Pregnancy rate (PR), pregnancy outcome. RESULTS: The overall PR in 118 cycles with luteal support was 26.3% whereas 10.4% pregnancies were achieved in 115 cycles of the control group. The influence of luteal support was more pronounced in patients with CC-resistant anovulation (25.2% versus 6.9%) than in patients with hypogonadotropic amenorrhea (33.3% versus 21.4%, not significant). The abortion rate was not significantly changed. CONCLUSION: Luteal support with P increases the PR after hMG and hCG induction. The need for supplementary P seems to be related to the underlying cause of ovarian disturbance. PMID- 8224263 TI - Comparison of growth hormone responses to growth hormone-releasing factor and clonidine in women with normal or poor ovarian response to gonadotropin stimulation. AB - OBJECTIVE: To assess the reliability of growth hormone (GH) secretion tests using provocative agents in women of different ovarian status. DESIGN: Comparison of GH secretion in response to clonidine (Catapressan; Boehringer Ingelheim, Reims, France) and growth hormone-releasing factor (GH-RF). SETTING: University Hospital, Hopital Tenon, Paris, France. PATIENTS: Women categorized as "normal" (n = 6) or "poor" responder (n = 7) to ovarian stimulation with gonadotropins, depending on the follicular development attained at previous IVF attempts. INTERVENTIONS: Clonidine (0.300 mg administered orally) once and GH-RF (1 micrograms/kg IV) repeated twice. The tests were performed in random order in each individual on following cycles. MAIN OUTCOME MEASURES: Basal and peak GH values, area under the curve (AUC). RESULTS: Poor responder patients show significantly higher basal levels of FSH, GH, and insulin-like growth factor 1; FSH and basal GH levels are positively correlated. Peak GH levels and AUC are not significantly different in both categories of patients, whether GH-releasing factor or clonidine are used as provocative agents. True positive rate is 56.4% at the cutoff value of 7 micrograms/L, with no significant difference between the patients of the normally or poorly responding groups. At the 10-micrograms/L cutoff level, the true positive rate is almost half in the poor responder group (19.0% versus 38.9%), but the difference is not significant. CONCLUSIONS: These results raise concern about using the GH secretory response to a single clonidine administration as a predictive test of the therapeutic benefit that could be obtained by co-stimulating the somatotropic axis during a treatment with gonadotropins in poor responder patients, especially when their FSH basal levels are elevated. PMID- 8224264 TI - Altered luteinizing hormone pulse frequency in early follicular phase of the menstrual cycle with luteal phase defect patients in women. AB - OBJECTIVE: To delineate the relationship between the pulsatile gonadotropin inputs in early follicular phase of the menstrual cycle and the P secretions by the corpus luteum in women. DESIGN: For measuring pulsatile release of gonadotropin, blood samples were drawn every 15 minutes for 24 hours in the early follicular phase. Daily blood samples were drawn for LH, FSH, E2, and P. SETTING: The reproductive endocrine unit of a university hospital. PATIENTS: Fourteen patients with luteal phase defect (LPD) and 12 normally cycling women. RESULTS: The length of follicular phase in LPD was significantly shorter than that of women with normal cycles. There were significant differences in LH pulsatile secretions and amplitudes in LPD patients when compared with those of women with normal cycles. Basal E2, PRL, and preovulatory E2 concentrations were not different between the two groups whereas the peak of P secretions in luteal phase was significantly decreased in LPD. CONCLUSIONS: These data suggest that LPD may result from the altered LH pulse frequency in early follicular phase of the menstrual cycle. Whether this increased LH pulse frequency results from an intrinsic disease of the pulse oscillator or to some event in the preceding cycle remains unknown. It is tempting to speculate that an increased LH pulsatile secretion in the early follicular phase of menstrual cycles in patients with LPD may down-regulate the LH secretion at midcycle, thereby lowering the LH surge, which in turn reduces the P secretion in luteal phase. PMID- 8224265 TI - Clinical and endocrine effects of flutamide in hyperandrogenic women. AB - OBJECTIVE: To investigate the clinical and endocrine effects of the antiandrogen flutamide in hirsute women. DESIGN: Hirsutism was assessed before and after 3 months of treatment with flutamide 500 mg/d. Endocrine evaluations were performed before and during the 2nd month of treatment with flutamide 500 mg or 750 mg/d. SETTING: Department of Obstetrics and Gynecology, Pisa, Italy. PARTICIPANTS: Eighteen hirsute women were studied: nine women were hyperandrogenic, and the other 9 had an idiopathic hirsutism. INTERVENTIONS: Women were randomly treated with flutamide 500 mg/d (9 patients) or 750 mg/d (9 patients) for 3 and 2 months, respectively. Six received placebo 1 month before flutamide treatment. MAIN OUTCOME MEASURES: Hirsutism was assessed by measuring hair diameter. Follicle stimulating hormone and LH responses to GnRH were evaluated. Basal plasma levels of T, androstenedione (A), 17-hydroxyprogesterone (17-OHP), DHEAS, cortisol (F), and sex hormone-binding globulin (SHBG) were evaluated. The same hormones were determined after a single dose of flutamide (250 or 500 mg) or placebo throughout a 12-hour period and in samples collected 60 and 120 minutes after ACTH intravenous injection. RESULTS: Hair diameter was reduced by 30%. Both dosages of flutamide did not change basal and stimulated gonadotropin, T, A, 17-OHP, F, and SHBG levels. Both dosages reduced stimulated DHEAS levels. CONCLUSIONS: Flutamide may have a beneficial effect on hirsutism. This effectiveness is mainly due to its peripheral antiandrogen action. However, an effect on the adrenal gland seems to be present. PMID- 8224266 TI - A new mutation within the deoxyribonucleic acid-binding domain of the androgen receptor gene in a family with complete androgen insensitivity syndrome. AB - OBJECTIVE: To study the androgen receptor gene in a large kindred with complete androgen insensitivity syndrome and positive receptor-binding activity. DESIGN: Enzymatic amplification coupled with single strand conformation polymorphism and DNA sequencing were used. RESULTS: In all the affected members, single strand conformation polymorphism showed a mobility shift in exon 2, suggesting a sequence alteration. Sequencing identified a valine into phenylalanine substitution at position 581 in the first zinc finger of the DNA-binding domain. CONCLUSIONS: This valine 581 is conserved in all nuclear receptors and has an important role for the recognition of the androgen response elements by the activated androgen receptor. This amino acid substitution has not been previously described in the androgen receptor in patients with androgen insensitivity syndrome. Moreover, single strand conformation polymorphism allowed determination of the generation in which the mutation appeared and performance of carrier diagnosis in this family. PMID- 8224267 TI - Hyperhomocysteinemia: a risk factor in women with unexplained recurrent early pregnancy loss. AB - OBJECTIVE: To establish the prevalence of hyperhomocysteinemia in women with unexplained recurrent early pregnancy loss. DESIGN: In a patient-control study, the methionine-homocysteine metabolism was investigated by a standardized oral methionine-loading test. SETTING: Gynecologic outpatient department of university hospital. PATIENTS: One-hundred and two women who had been referred to the hospital because they suffered from at least two consecutive unexplained spontaneous abortions (study group) as well as 41 controls who were recruited by public advertisement were selected. INTERVENTIONS: Blood samples were collected just before and 6 hours after oral methionine administration to determine plasma total homocysteine concentrations. MAIN OUTCOME MEASURE: Plasma total homocysteine concentrations 6 hours after methionine loading. Hyperhomocysteinemia was defined as total homocysteine concentration at 6 hours exceeding the 97.5 percentile level of the controls. RESULTS: Hyperhomocysteinemia was diagnosed in 21 women of the study group (21%). In the parous women of the study group, the prevalence of hyperhomocysteinemia was more than two times greater compared with the nulliparous subjects (33% and 14%, respectively). CONCLUSION: Hyperhomocysteinemia is a risk factor in women with unexplained recurrent early pregnancy loss. PMID- 8224268 TI - Elevated levels of interleukin-6 in ascites and serum from women with ovarian hyperstimulation syndrome. AB - OBJECTIVE: To examine the role of pro-inflammatory cytokines in ovarian hyperstimulation syndrome (OHSS). DESIGN: In vitro laboratory study of serum, peritoneal cells isolated and fluid obtained from ascites removed in the therapeutic management of four patients with OHSS. SETTING: Tertiary care referral teaching hospital. PATIENTS: Four patients with OHSS comprised the study population. Five healthy women at the time of elective laparoscopic tubal ligation served as controls. Control serum was also obtained from healthy adult volunteers, and control peritoneal fluid (PF) was obtained from patients on peritoneal dialysis. INTERVENTIONS: Therapeutic paracentesis was performed on four patients with OHSS. RESULTS: Peritoneal cells were isolated and fluid obtained from ascites removed in the therapeutic management of the women with OHSS. Peritoneal cells were obtained by intraoperative lavage from the control women. The cells were incubated with various concentrations of Escherichia coli lipopolysaccharide (LPS) for 20 and 48 hours. Interleukin-1 beta (IL-1 beta), tumor necrosis factor (TNF), interleukin-6 (IL-6), and prostaglandin E2 (PGE2) were assayed in the incubation supernatants. The release of the three cytokines and PGE2 in response to LPS by peritoneal cells from women with OHSS was not different from the controls. However, both serum and ascitic fluid from women with OHSS contained significantly greater levels of IL-6 than control serum and PF. No significant differences in TNF levels in serum, ascitic fluid, or PF could be found by ELISA or bioassay. CONCLUSIONS: Increased production of IL-6, released into the peritoneal cavity and the circulation, may mediate OHSS. PMID- 8224269 TI - Expression of major histocompatibility complex class II antigen in endometriotic tissue in patients with endometriosis and adenomyosis. AB - OBJECTIVE: To evaluate the expression of major histocompatibility complex class II antigen (human leukocyte antigen, HLA-DR) in glandular cells in eutopic and ectopic endometria in patients with endometriosis and adenomyosis. PATIENTS: Subjects consisted of 50 patients with adenomyosis and 18 patients with endometriosis. Controls consisted of 17 patients with adenomyoma and 42 infertile women without habitual abortion and autoimmune diseases. DESIGN: Glandular cells in eutopic and ectopic endometria were analyzed immunohistochemically to localize the expression of HLA-DR antigen. The sections were scored from 0 to 5 according to the evaluation nomogram. RESULTS: Immunohistochemical analysis revealed significantly increased expression of HLA-DR antigen in the glandular cells in eutopic and ectopic endometria, compared with materials from patients with adenomyoma and infertile patients. CONCLUSIONS: It is likely that aberrant expression of HLA-DR antigen in glandular cells of eutopic and ectopic endometria in endometriosis and adenomyosis is deeply involved in various immunological abnormalities. PMID- 8224270 TI - Follicle-stimulating hormone beta gene structure in premature ovarian failure. AB - OBJECTIVE: To determine whether mutations in the gene for FSH beta are present, and possibly etiologic, in some patients with 46,XX premature ovarian failure (POF). DESIGN: DNA samples obtained from 18 study patients with POF and two menopausal fertile controls were studied by Southern blot analysis. DNA sequencing was performed in one patient. SETTING: Patients were seen in a reproductive endocrinology clinic and studied in a medical school laboratory setting at the Medical College of Georgia and Tufts University. MAIN OUTCOME MEASURES: Restriction fragment sizes on autoradiographs were compared between the study group and controls. DNA sequencing radiographs were compared between one study patient and five controls. RESULTS: Fragment sizes obtained with the restriction enzymes EcoRI, DraI, HincII, PstI, KpnI, BglI, BamHI, and BglII were similar size in both study subjects and controls using the probes pFSH beta-1.4 and pFSH beta-1.2. A previously described HindIII polymorphism was present using pFSH beta-1.2, but HindIII fragment sizes were identical in patients with ovarian failure and controls using pFSH beta-1.4. DNA sequencing of the FSH beta gene in one patient was normal. CONCLUSIONS: No mutations in the gene for FSH beta were identified in women with POF. DNA sequencing of the exons and promoter region of the FSH beta gene in one woman with POF was normal. This does not entirely exclude the possibility that smaller deletions, insertions, or point mutations of the FSH beta could be etiologic in some women with POF. The HindIII polymorphism does not appear to segregate with 46,XX POF. PMID- 8224271 TI - In vitro fertilization and embryo transfer for the treatment of infertility associated with polycystic ovary syndrome. AB - OBJECTIVE: To examine the outcome of treatment with IVF-ET of women with polycystic ovarian syndrome (PCOS) who failed to conceive on conventional treatment. DESIGN: Retrospective analysis with an age-matched control group. SETTING: University hospital infertility clinic and IVF unit. PATIENTS. INTERVENTIONS: Sixty-eight women with PCOS who had failed to conceive on treatment with clomiphene citrate and during six ovulatory cycles on gonadotropins underwent 208 cycles of IVF-ET. An age-matched group of 68 women with a tubal mechanical factor who received 143 treatment cycles during the same period served as controls. MAIN OUTCOME MEASURES: Cumulative conception rates, the cumulative livebirth rates, and IVF-ET data were compared between the two groups. Results of treatment with and without GnRH agonist (GnRH-a) within the groups were also compared. RESULTS: A comparison of PCOS and mechanical factor (control) groups showed almost identical results at 6 months for cumulative conception rate (82% versus 85%) and cumulative livebirth rate (69% versus 65%). Significantly more oocytes were retrieved but a smaller percentage fertilized in PCOS, and the pregnancy rate per ET did not differ between the two groups (23% versus 26%). Treatment with GnRH-a and gonadotropins as opposed to gonadotropins alone improved the cumulative conception rate, miscarriage rate, and cumulative livebirth rate in the PCOS but not in the control group and improved fertilization rates in both groups. CONCLUSIONS: For patients with PCOS who fail to conceive with gonadotropin treatment, IVF-ET is a successful treatment alternative, producing results equal to those for women with a mechanical tubal factor. Better results were achieved with GnRH-a in women with PCOS but made no difference to those with a mechanical tubal factor compared with treatment with gonadotropins alone. PMID- 8224272 TI - A novel ovarian stimulation protocol for use with the assisted reproductive technologies. AB - OBJECTIVE: To determine whether a new ovarian stimulation protocol termed "minimal stimulation" provides pregnancy rates (PRs) comparable with those in a conventional full stimulation protocol for patients undergoing assisted reproductive technologies (ART). DESIGN: Prospective, nonrandomized study of patients in minimal stimulation or full stimulation, followed by standard IVF and zygote intrafallopian transfer or uterine-ET. SETTING: The ART program of the Mayo Clinic, Rochester, Minnesota. PATIENTS: Women (n = 120) 42 years of age or younger with serum day 3 FSH level < or = 15.0 mIU/mL (conversion factor to SI unit, 1.0), normal thyroid-stimulating hormone and PRL levels, normal endometrial cavity as observed on hysterosalpingogram, and partners with a normal semen analysis. INTERVENTIONS: Oral clomiphene citrate, hMG, oocyte retrieval, IVF-ET. MAIN OUTCOME MEASURES: Cancellations, implantation, pregnancy. RESULTS: The cancellation rate was not significantly higher in minimal stimulation (25.8%) than in full stimulation (14.1%). Minimal stimulation cycles yielded fewer oocytes per aspiration (3.4 +/- 1.6) than full stimulation (10.1 +/- 5.4). There was no difference in the implantation rates per ET (minimal stimulation, 16.4%; full stimulation, 13.3%) or overall clinical PRs per retrieval (minimal stimulation, 31%; full stimulation, 42%). CONCLUSIONS: Minimal stimulation for IVF is less expensive than full stimulation and minimizes monitoring and patient discomfort. In addition, it produces acceptable PRs and represents an attractive alternative to select patients undergoing ART. PMID- 8224273 TI - Assisted hatching facilitates earlier implantation. AB - OBJECTIVE: To examine whether opening of the zona pellucida (i.e., assisted hatching) accelerates implantation. DESIGN: In a controlled, randomized trial, patients were assigned to control and assisted hatching groups. SETTING: All patients studied were of the Center for Reproductive Medicine at Cornell University Medical College. INTERVENTION: All patients underwent stimulation with gonadotropins after luteal phase GnRH down regulation. Assisted hatching with zona drilling using acidic Tyrode's solution was performed on the assigned embryos. MAIN OUTCOME MEASURES: Luteal E2, P, and hCG on days +5, +6, +7, +8, +9, +11, +13, and +15 were measured. The implantation time, peak midluteal E2 and intervals between these two values were studied. RESULTS: Implantation occurred significantly earlier in the assisted hatching group. The interval between implantation and peak midluteal E2 was also significantly shorter in the assisted hatching group than in the controls. However, there was no significant difference in the day of the peak midluteal E2 between the assisted hatching and control groups. CONCLUSION: Assisted hatching may enhance embryo implantation not only by mechanically facilitating the hatching process but also by allowing earlier embryo-endometrium contact. Such early contact may enhance embryonic development potential and may optimize synchronization between embryo and endometrium, resulting in improved implantation efficiency. PMID- 8224274 TI - In vitro deleterious effect of hypoxanthine in Ham's Nutrient Mixture F-10 culture medium on human oocyte fertilization and early embryonic development. AB - OBJECTIVE: To determine whether hypoxanthine in Ham's Nutrient Mixture F-10 (GIBCO, Grand Island, NY) culture medium affects murine embryo development or the outcome of patients undergoing IVF-ET. DESIGN: For the two-cell embryo bioassay, embryos from each female were equally distributed and incubated in Ham's F-10 with or without hypoxanthine supplementation for up to 72 hours. To assess the effect of hypoxanthine in human IVF-ET, oocytes, sperm, and embryos were cultured in Ham's F-10 medium with or without hypoxanthine. Fertilization and embryo cleavage were assessed at 18 and 40 hours, respectively, after insemination. SETTING: University medical research laboratory. PATIENTS, PARTICIPANTS: Nine couples undergoing IVF-ET. RESULTS: Two-cell mouse embryos incubated for 24 hours without hypoxanthine developed 40% to morula, compared with 6.5% for the hypoxanthine group. At 72 hours, 99.5% of the embryos without hypoxanthine reached the expanded blastocyst stage with 65% of them exhibiting hatching, compared with 72% and 19.5%, respectively, for the group with hypoxanthine. Human oocytes cultured in Ham's F-10 without hypoxanthine showed higher fertilization rates than the group incubated in the presence of hypoxanthine (69% versus 53%). The proportion of cleaved embryos was not different between the two culture media; however, the rate of embryos cleaving without cytoplasmic fragments was significantly higher in the group without hypoxanthine (75% versus 35%). CONCLUSION: Ham's F-10 medium containing hypoxanthine significantly reduced embryo development in the two-cell mouse embryo bioassay. Hypoxanthine in culture medium for IVF-ET may have a deleterious effect on human gametes, leading to decreased fertilization and increased incidence of cytoplasmic fragments in the conceptuses. PMID- 8224275 TI - Is spermatozoan acrosin a predictor of fertilization and embryo quality in the human? AB - OBJECTIVE: To investigate whether acrosin is a more reliable criterion than conventional parameters for assessing semen samples. DESIGN: Total acrosin was estimated biochemically in semen samples obtained for routine screening for infertility and for IVF-ET procedures. SETTING: Assisted Conception Unit, St. James's University Hospital, Leeds, United Kingdom. PATIENTS: Four hundred sixty three couples being investigated for causes of infertility, and 132 couples undergoing IVF-ET for any indication except antisperm antibodies between 1990 and 1991 were included in the study. INTERVENTIONS: Semen samples were collected as part of routine investigation. Samples from men with consistently high viscosity were collected in alpha-chymotrypsin. MAIN OUTCOME MEASURES: Total spermatozoan acrosin in motile spermatozoa and motile spermatozoan density in couples being assessed for IVF-ET and fertilization and embryo quality in those receiving treatment are considered. RESULTS: Total acrosin levels were less variable (within subject) than either total or motile spermatozoan concentration at ejaculation. Although severely oligozoospermic ejaculates had the lowest levels of total acrosin, overall, there was no significant correlation of spermatozoan concentration between total acrosin levels and percentage fertilization. CONCLUSIONS: Total spermatozoan acrosin activity correlates positively with fertilization rates but not with spermatozoan count. Motile spermatozoan density for insemination may be adjusted to achieve > 7.5 microIU acrosin per oocyte, without compromising fertilization or further embryo development to blastocysts in vitro. PMID- 8224277 TI - Effects of peritoneal fluids from patients with endometriosis on capacitated spermatozoa. AB - OBJECTIVE: To determine the effects of peritoneal fluid (PF) from patients with endometriosis on capacitated sperm. DESIGN: Capacitated donor sperm were allowed to migrate into straws filled with pooled PF from patients with unexplained infertility without endometriosis (n = 4) and those with mild (n = 4) and moderate-severe (n = 4) endometriosis, respectively, for 3 hours. MAIN OUTCOME MEASURES: Sperm motility, sperm membrane integrity, and acrosomal loss were determined. RESULTS: Statistically, no significant differences were found among the various sperm parameters determined. CONCLUSION: Peritoneal fluid from patients with endometriosis does not appear to adversely affect capacitated spermatozoa. PMID- 8224276 TI - Influence of sperm parameters on embryo quality. AB - OBJECTIVE: To evaluate the influence of sperm defects on embryo quality. DESIGN: Retrospective study. SETTING: In vitro fertilization center. PATIENTS: Embryo transfers (710) from IVF attempts for tubal disease (626) or male infertility (84). MAIN OUTCOME MEASURES: Embryo morphology as a function of causes of infertility, semen, and follicular growth parameters. Embryos were classified into three groups according to their morphology. RESULTS: Transfers of embryos with good morphology were associated to a higher pregnancy rate (34%) than those with intermediate (24%) and poor (10%) morphology. Transfers of embryos with a poor morphology were more frequent (26 of 84 versus 114 of 626) and those with a fair aspect were less frequent (24 of 84 versus 229 of 626) in male infertility than in tubal disease. Embryos with a poor morphology were associated with lower percentage of morphologically normal sperms (62% +/- 19% versus 67% +/- 18%; means +/- SD) and a higher percentage of abnormalities of the postacrosomial region (29% +/- 15% versus 18% +/- 7%). Moreover, sperms with counts < 10 x 10(6)/mL were associated with a lower percentage of embryos with good morphology (18% versus 37%) than sperms with counts > or = 10 x 10(6)/mL. CONCLUSION: Embryo quality is influenced by the semen quality and especially by sperm head abnormalities, suggesting an important role of the male gamete on the early stages of embryogenesis. PMID- 8224278 TI - Effect of epidermal growth factor on human sperm capacitation. AB - OBJECTIVE: To examine the effect of epidermal growth factor (EGF) on human sperm capacitation and the involvement of protein phosphorylation in the regulation of the EGF action. DESIGN: Human sperm were capacitated in modified Krebs-Ringer's bicarbonate medium of Biggers, Whitten, and Whittingham in the presence of EGF and various agents known to phosphorylate the EGF receptor. The chlortetracycline fluorescence assay was used to monitor capacitated sperm. Capacitation was confirmed by the ability of sperm to undergo the acrosome reaction in response to solubilized mouse zonae pellucidae (ZP). RESULTS: In 15 minutes, the appearance of the clear perimeter pattern increased significantly in the sperm treated with 100 ng/mL EGF compared with the controls. In EGF-treated sperm, the percent clear perimeter pattern remained stable for 3 hours without affecting the acrosome reaction pattern and the motility. Epidermal growth factor stimulated the appearance of the clear perimeter pattern at concentrations > 100 pg/mL. The stimulation by EGF was attenuated by the treatment with genistein, 12-O tetradecanoylphorbol-13-acetate, or thapsigargin. In sperm that were incubated in the presence of 100 ng/mL EGF for 30 minutes and further induced the acrosome reaction by mouse ZP, the percent acrosome reaction pattern increased significantly compared with the controls. CONCLUSION: Epidermal growth factor stimulates human sperm capacitation by activating the tyrosine kinase of the EGF receptor which is regulated by multisite phosphorylation. PMID- 8224279 TI - Cryopreservation of human spermatozoa. IV. The effects of cooling rate and warming rate on the maintenance of motility, plasma membrane integrity, and mitochondrial function. AB - OBJECTIVE: To test the hypotheses that there is a two-factor aspect of cellular damage during cryopreservation that occurs in human sperm (osmotic effects versus intracellular ice formation) and that there is a cooling rate by warming rate interaction related to this damage. DESIGN: Ejaculates from healthy men were cooled at 0.1, 1.0, 10, 175, or 800 degrees C/min to -80 degrees C in a solution of 0.85 M glycerol and plunged into liquid nitrogen. Samples were warmed at 400 degrees C/min (experiment 1) or either 1 degrees C or 400 degrees C/min (experiment 2). After warming, sperm were assessed for survival using motility as the endpoint in experiment 1 and motility, plasma membrane integrity, and mitochondrial function in experiment 2. RESULTS: In experiment 1, over the various cooling rates with a standard 400 degrees C/min warming rate, a plot of motility versus cooling rate produced a classical inverted U-shaped curve (n = 6) with maximum motility at the 10 degrees C/min cooling rate. In experiment 2, over the various cooling rates, both 1 and 400 degrees C/min warming rates produced similar but shifted plots of motility, plasma membrane integrity, and mitochondrial function versus cooling rate, which also produced inverted U-shaped patterns (n = 11). Maximal survival for each of the three endpoints occurred at 10 degrees C/min cooling rate for the rapidly warmed sperm and at 1 degree C/min for the slowly warmed sperm. CONCLUSIONS: These data support the hypotheses that a two-factor hypothesis of cryodamage applies to human spermatozoa and that an interaction exists between cooling rate and warming rate. These data also suggest that motility, plasma membrane integrity, and mitochondrial function are not differently affected by cooling and warming during cryopreservation. PMID- 8224280 TI - Embryo transfer technique as a cause of ectopic pregnancy in in vitro fertilization. AB - A randomized prospective study was performed to compare the effects of a midfundal versus a deep fundal transfer technique on subsequent intrauterine and ectopic PRs after IVF. The clinical intra-uterine PR after the deep fundal transfer was 12.4% per cycle with a 1.5% ectopic PR (which represented 12.2% of the pregnancies) versus 14.2% IUPs per cycle with a 0.4% ectopic rate (representing 3% of pregnancies) after midfundal transfers. The midfundal technique appears superior to deep fundal procedures because of a lower percentage of EPs without any sacrifice of the intrauterine PR after midfundal transfers. PMID- 8224281 TI - Tympanic thermometry for recording basal body temperatures. AB - Evaluation of 12 menstrual cycles using oral, rectal, and TM temperature measuring devices (over 2,000 individual readings) confirmed the ovulatory thermal shift was equally detected with TM thermometry compared with the traditional methods. Although a single TM reading was satisfactory, an average of three successive readings provided a smoother graph (decreased variance). The device appears acceptable, and even preferred, for recording BBT charts, primarily because of its nearly instantaneous readings. PMID- 8224282 TI - The "darkness hormone". PMID- 8224283 TI - "Killer cells" and endometriosis. PMID- 8224284 TI - Risks of intratubal methotrexate. PMID- 8224285 TI - Risks of intratubal methotrexate. PMID- 8224286 TI - Edridge-Green Lecture. Competition and cooperation in visual development. AB - Studies of the effect of visual deprivation on cells in the lateral geniculate nucleus (LGN) show that there are two distinct sensitive periods in the monkey during which different reactions between the visual pathways related to the two eyes predominate and requirements for recovery from deprivation differ. The first extends from birth to about 8 weeks of age. The main interaction between the pathways from the two eyes is competitive, segregation of cortical ocular dominance columns occurs during this early period and monocular deprivation results initially in hypertrophy of undeprived LGN cells, with later parallel shrinkage of both deprived and undeprived parvocellular cells. Simply reopening the closed eye produces no recovery but reverse suture is effective in reversing some of the changes. The second sensitive period starts from about 8 weeks of age, although the peak of the later sensitivity appears to be at 7-9 months of age and some effect is still present at 12-18 months. During this later phase a cooperative interaction between the pathways related to the two eyes is necessary for normal development and in the absence of this selective shrinkage of both deprived and undeprived parvocellular LGN cells occurs. Simply reopening an eye during this late sensitive period allows recovery of these cells to normal size. PMID- 8224287 TI - Strabismus: challenges and trends. PMID- 8224288 TI - The outcome of strabismus surgery in childhood esotropia. AB - The results of squint surgery in 118 children with non-paralytic childhood esotropia are analysed. A 'favourable outcome', defined as a final alignment within +/- 10 dioptres of straight, or within +/- 20 dioptres of straight if there was evidence of binocular single vision, was achieved in 86 (72.9%) children. The factors affecting the final outcome are discussed, including age of onset, age at the time of surgery, pre-operative and post-operative amblyopia, refractive error, anisometropia, the surgical procedures used, and post-operative ocular alignment. PMID- 8224289 TI - Subconjunctival bupivacaine in strabismus surgery. AB - A prospective study was performed to assess the effect of subconjunctival bupivacaine on pain relief following strabismus surgery. A total of 36 children were randomly allocated to receive either subconjunctival infiltration of bupivacaine or normal saline at the conclusion of surgery. Post-operative pain was evaluated using a 4-point assessment score. Subconjunctival bupivacaine provided significantly better post-operative pain relief (P < 0.001). We suggest that it could be used routinely in strabismus surgery, as it improves post operative comfort thereby facilitating day case surgery. PMID- 8224290 TI - Ophthalmic herpes zoster. AB - A current review of ophthalmic zoster is presented including its virology, immunology, epidemiology and pathogenesis. We give our findings in 1356 patients referred to the Zoster Clinic at Moorfields Eye Hospital, London. The treatment of the disease and its ocular complications is discussed. PMID- 8224291 TI - Chronic progressive conjunctival cicatrisation. AB - The aim of this review is to demonstrate the spectrum of conditions encompassed by the term 'chronic progressive cicatrising conjunctivitis', to discuss mechanisms of conjunctival scar tissue formation and to describe the sequelae and therapeutic options in this potentially blinding condition. Chronic progressive cicatrising conjunctivitis is found in association with some mucocutaneous disorders (cicatricial pemphigoid, linear IgA disease), as part of paraneoplastic syndromes and after long-term treatment with certain systemic and topical medications (pseudo-pemphigoid). Recent studies on the conjunctiva of pemphigoid patients indicate that macrophages may play a pivotal role in chronic progressive conjunctival cicatrisation. They mediate the transition from inflammation to scar tissue by secretion of fibrogenic cytokines. There is evidence that similar mechanisms are involved in the other fibrosing conjunctival disorders. Sequelae of chronic conjunctival cicatrisation include the obstruction of lacrimal and meibomian glands, tear film alterations, trichiasis, keratopathy and blindness. Present possibilities and future options for the treatment of this condition are discussed. PMID- 8224292 TI - Ultrastructural study of a corneal keloid. AB - A 69-year-old man developed keloid overgrowth on his left cornea in response to an injury from a fingernail. The lesion was removed by superficial lamellar keratectomy and was studied by electron microscopy, and light microscopy after immunoperoxidase staining for actin. The surgical specimen revealed disorganised, anteriorly atrophied and posteriorly vascularised connective tissue stroma. The epithelium was oedematous, thin, non-keratinised, and contained cells with features of myoblastic differentiation. Stromal fibroblasts were found in several distinct ultrastructural forms including young active fibroblasts, myofibroblasts, inactive fibroblasts (fibrocytes) and fibroblasts with prominently fibrillar cytoplasm. Fibroblasts with glycogen storage and/or pseudonuclear inclusions were also seen. Macrophages and lymphocytes were scattered in the stroma, and intact nerves were also present. An irregular 2-65 microns band of 10 nm filament meshwork existed at the posterior border of the keloid stroma, and deep localised patches of climatic degeneration were detected. PMID- 8224293 TI - An approach to the management of patients with uveal melanoma. PMID- 8224294 TI - Peripapillary subretinal neovascular membranes: the natural history. AB - We studied the natural course of peripapillary subretinal neovascular membranes in 20 eyes. The membranes were either idiopathic in nature or associated with underlying disease. We found that the visual prognosis was related to the behaviour of the membrane and to the patient's age. Patients under the age of 40 years had unilateral disease and a more favourable visual prognosis. Older patients had a uniformly poor visual prognosis and a high incidence of bilateral disease. Fifty-four per cent of patients over 45 years and 62% over 70 years of age had bilateral membranes. In those over the age of 70 years, 75% of untreated eyes lost vision to a level of 3/60 or less in both eyes. The probability of the second eye becoming involved in the older group of patients was 62%. The period of time elapsing between involvement of the first and second eye varied between simultaneous occurrence and a delay of 7 years. PMID- 8224295 TI - Doppler ultrasonography of the central retinal artery in normals treated with topical timolol. AB - Using high-resolution Duplex scanning it is possible to evaluate the blood flow velocity in the central retinal artery of eyes of normal individuals. The flow velocity can be divided into its systolic and diastolic phases. Topical timolol maleate 0.5%, a non-selective beta-adrenergic receptor blocking agent, was then administered to one eye in each of the normals and the flow velocity was again recorded. The flow velocity improvement was significant, with a 43.58% increase in the systolic phase and a 61.53% increase in the diastolic phase. The diastolic component increased from 49.29% to 55.56%. The increased flow velocity may be due to a vasodilatory effect of timolol. The technique is briefly described and the significance of the results discussed. PMID- 8224296 TI - Macular ischaemia in posterior uveitis. AB - The commonest cause of visual morbidity in patients with posterior uveitis is cystoid macular oedema, which usually responds to immunosuppressive treatment. However, a small group of patients do not have a satisfactory visual outcome despite apparently adequate therapy. In a retrospective study of 345 angiograms of 135 patients with active non-occlusive retinal vasculitis 12 patients were identified by independent masked review as showing macular ischaemia on their fluorescein angiograms. Four patients had Behcet's disease, 4 sarcoidosis, and 4 idiopathic retinal vasculitis. Follow-up of these patients for an average of 36 months (range 6-120 months) showed that visual acuity failed to improve in 4 patients and dropped by an average of three lines Snellen in the other 8. We suggest that a poor visual outcome in some patients with posterior uveitis may be predicted by the presence of macular ischaemia on fluorescein angiography and that immunosuppressive therapy should be prescribed with caution in these patients. PMID- 8224297 TI - The spectrum of ocular toxocariasis. AB - Ocular toxocariasis is rare and therefore the spectrum of clinical disease is difficult to establish. We present a review of the clinical features and laboratory findings in a group of patients with positive Toxocara serology and ocular toxocariasis. The clinical spectrum was diverse and milder disease was commoner than might be supposed from reviews of the literature. Eosinophilia was unusual, but featured in two cases of unilateral pars planitis. PMID- 8224298 TI - Ocular siderosis. AB - The authors report their experience in the management of 8 patients with ocular siderosis due to a retained intraocular foreign body (IOFB). All patients were male, aged between 19 and 39 years. Seven had a definite history of trauma; 3 had presented at the time of injury to a casualty department, and the diagnosis had been missed. The interval between injury and diagnosis ranged from 2 to 24 months. IOFB removal was performed in 7 patients: through a sclerotomy and magnet or foreign body forceps in 4 eyes and via a pars plana vitrectomy and intraocular foreign body forceps in 3 eyes. Cataract extraction was performed in 4 patients. Histological examination of specimens removed at the time of surgery showed iron deposition in the conjunctiva, anterior lens capsule and pars plana. Transmission electron microscope X-ray microanalysis showed that iron was contained in siderosomes, intracytoplasmic membrane-bound dense bodies. Final visual acuity was 6/12 or better in 6 patients and reduced to light perception in the remaining 2 due to proliferative vitreoretinopathy. PMID- 8224299 TI - Fuchs' heterochromic cyclitis and retinal vascular abnormalities in progressive hemifacial atrophy. AB - We report a case of progressive hemifacial atrophy with a combination of ipsilateral Fuchs' heterochromic cyclitis and retinal vascular abnormalities. Recently, evidence was found for a neurovascular defect in hemifacial atrophy. Our case not only supports the (clinical) association between Fuchs' heterochromic cyclitis and hemifacial atrophy, but the retinal vascular abnormalities found in this patient add further support to the existence of a neurovascular defect. These findings and our short review of the literature point to the hypothesis of a common sympathetic defect, implicated in the aetiology of both Fuchs' heterochromic cyclitis and progressive hemifacial atrophy. It has to be borne in mind, however, that Fuchs' heterochromic cyclitis has been reported in association with other diseases. It seems likely that although Fuchs' heterochromic cyclitis is a single clinical entity, it may have more than one cause. PMID- 8224300 TI - Penetrating eye injuries in children of the West Bank and Gaza strip. AB - A retrospective study was carried out on 118 children with penetrating eye injuries. Fifty-three per cent of those without an intraocular foreign body (50/94) achieved a final visual acuity of 6/12 or better, whilst 13% (14/108) required an enucleation. Mixed corneoscleral wounds, vitreous haemorrhage and retinal detachment were bad prognostic features. Important principles of management are discussed. PMID- 8224301 TI - Patient satisfaction following vitreoretinal surgery. AB - A postal questionnaire was used to study patient satisfaction in 149 consecutive patients undergoing vitreoretinal surgery in Southampton. Of the 123 patients who responded, 70 (60%) felt that their vision improved following surgery. Fifty eight patients (47%) stated that their vision was not as good as they had hoped it would be, and multiple logistic regression analysis showed that this response reflected a poorer visual outcome in this group. Despite this, 116 (94%) of those replying stated that the operation had been worth while. All 5 patients who felt that their operations had not been worth while had undergone retinal detachment surgery. This high level of satisfaction may be attributable to realistic expectations, as 116 patients (94%) replied that the procedures had been adequately explained. PMID- 8224302 TI - Ab interno pulsed dye laser sclerostomy for the treatment of glaucoma: preliminary results of a new technique. AB - Ab interno gonioscopic laser sclerostomy in conjunction with iontophoretic staining of the sclera was performed on 12 patients with end-stage glaucoma in order to ascertain the feasibility of the technique. In 9 of 12 patients (75%) a visible sclerostomy was formed and mean intraocular pressure (IOP) dropped from 36.3 mmHg prior to treatment to 25.3 mmHg at 1 week, a fall of 30.3%. The drop in IOP was maintained throughout a 3-month follow-up period, with 6 of 11 patients having IOP controlled below 21 mmHG. The procedure is described together with our preliminary results, and possible advantages and areas of improvement for the technique are discussed. PMID- 8224303 TI - Trabeculectomy: effect of varying surgical site. AB - Trabeculectomy reduces intraocular pressure by fistula formation into a subconjunctival bleb. The operation site traditionally has been at the superior corneo-scleral junction with a failure rate of 10-30%. The causes of trabeculectomy failure can be broadly classified into intraocular, scleral and extraocular. Extraocular factors account for the majority of failures, the main cause being increased subconjunctival fibrosis. Hitherto the effect of varying the site of filtration surgery on intraocular pressure control has not been studied. We therefore carried out a prospective study to evaluate the optimum site for trabeculectomy. Sixty patients were randomised to undergo a superior, nasal or temporal trabeculectomy. Patients who underwent a nasal trabeculectomy had significantly lower intraocular pressures 18 months after surgery (p < 0.05), with 30% having an intraocular pressure of < or = 10 mmHg. PMID- 8224304 TI - The long-term follow-up of patients treated with YAG laser to re-open closed or closing fistulae following glaucoma surgery. AB - The Q-switched neodymium:YAG laser was used to reopen closed or closing fistulae following surgery for glaucoma. Before laser treatment the cause of the obstruction of the fistula was located using a gonioscope. Following laser treatment a significant lowering of the intraocular pressure was seen with minimal complications, which has been maintained over a period of 5 years. PMID- 8224305 TI - Refractive results after corneal triple procedures (PK+ECCE+IOL) AB - We have analysed the refractive results of corneal triple procedures (penetrating keratoplasty with extracapsular cataract extraction and posterior chamber intraocular lens implantation) in 52 eyes of 47 patients with a mean follow-up of 39 months. The patients were predominantly old and female and most received unilateral surgery. The contralateral acuity was 6/24 or worse in more than 50% of cases at the time of surgery. Many of these patients were significantly ametropic pre-operatively. Biometry does not seem to have improved the refractive results in those patients in whom it was attempted. The majority of patients were hypermetropic when their refraction stabilised, with resulting poor uncorrected visual performance. Possible improvements are discussed. PMID- 8224306 TI - Inter-observer variation in clinical optic disc biometry. AB - This study investigated the inter-observer reproducibility of measurements of the optic nerve head as carried out with a clinical optic disc biometer. This instrument employs a modification of indirect ophthalmoscopy to enable measurement of fundus structures. Measurements were made independently by two observers on 84 eyes of 47 patients. The median inter-observer differences for each measurement were as follows: maximum disc diameter, 0.085 mm; minimum disc diameter, 0.080 mm; maximum cup diameter, 0.098 mm; minimum cup diameter, 0.078 mm; disc area, 0.225 mm2; neuroretinal rim area, 0.215 mm2. Utilising the above measurements, the instrument automatically calculates a 'rim index' to take account of variations in disc size. It then interprets the disc as normal, suspicious or glaucomatous. The optic disc biometer produced seriously conflicting interpretations in 8 discs, 7 of which had been judged by both observers to have indistinct boundaries of the disc or cup. PMID- 8224307 TI - Conjunctival changes after subconjunctival lignocaine. AB - Subconjunctival injection of 2% lignocaine at the 12 o'clock position was used as the local anaesthetic in 19 eyes undergoing a primary trabeculectomy for open angle glaucoma. The appearance of the bleb and mean post-operative intraocular pressure (IOP) were compared with those in 29 eyes with the same diagnosis undergoing trabeculectomy under general anaesthesia by the same surgeons over the same time period. The two groups of eyes were similar with regard to treatment with beta blockers (p > 0.1), miotics (p > 0.25), sympathomimetics (p > 0.25), carbonic anhydrase inhibitors (p > 0.5), or no treatment (p > 0.25). Seventy seven per cent of the local anaesthetic group and 25% of the general anaesthetic group developed avascular, thin-walled drainage blebs (p < 0.001). The mean post operative IOP was significantly lower in the group receiving local anaesthetic (p < 0.001). The reasons for and significance of these observations are discussed, and the merits and disadvantages of thin-walled blebs are evaluated. PMID- 8224308 TI - Antigen sensitivity evaluated by tear-specific and serum-specific IgE, skin tests, and conjunctival and nasal provocation tests in patients with ocular allergic disease. AB - The potential for ocular allergic patients to have a site-specific antigen sensitisation was investigated using various diagnostic tests of allergen sensitivity in subjects with allergic conjunctivitis (AC: n = 135), vernal keratoconjunctivitis (VK: n = 20), rhinoconjunctivitis (n = 20) or rhinitis (N = 10). In the AC and VK patients, skin tests and conjunctival provocation tests (CPT) were performed, and the levels of specific IgE in serum and in tears were identified. A subgroup of 36 patients was also challenged with a nasal-specific provocation test (NPT). Results showed a poor correlation between skin test results and tear-specific IgE, and also between serum-specific IgE and tear specific IgE in both AC and VK patients (K < 0.3). CPT and tear IgE were significantly correlated (K = 0.5) in the ocular allergic population. In patients with rhinoconjunctivitis or rhinitis, and in 10 normal subjects, results of CPT and NPT were in 100% agreement. Conversely, in patients with only conjunctivitis, little correlation was found between the results of CPT and NPT (K = 0.3). Tear specific IgE was the only positive diagnostic sign of antigen sensitivity in 35% of VK patients and 30% of AC patients. These results suggest that the conjunctiva can be a uniquely sensitised target organ in allergic patients. PMID- 8224309 TI - Iris prolapse; who? When? Why? AB - A 12-month period was reviewed to identify the incidence of iris prolapse following cataract surgery, and any predisposing factors. Of 1408 routine manual extracapsular cataract extractions, 29 eyes (2.06%) sustained an iris prolapse. Iris prolapse was commoner in Asian patients and when less experienced surgeons were operating. Twenty-five per cent of cases occurred in patients with obstructive airways disease or post-operative cough. Thirty-four per cent of iris prolapses were identified on the first post-operative day and 86% within 2 weeks. Prolapse probably occurs because of raised intraocular pressure in association with poor wound construction or closure. More emphasis should be given to wound construction and closure during surgical training, with extra diligence when operating on patients likely to cough postoperatively. Clinicians must ensure non English speaking patients understand about ocular aftercare following cataract surgery. PMID- 8224310 TI - No eye pad for corneal abrasion. AB - We have carried out a randomised clinical trial to assess the healing rate and level of discomfort experienced in two groups of patients with simple traumatic corneal abrasions. Patients treated with antibiotic ointment and mydriatic alone have a significantly improved healing rate compared with those treated with antibiotic ointment, mydriatic and a double eye pad with bandage (0.05 > p > 0.02). There was no significant difference in the level of discomfort experienced by the two groups. PMID- 8224311 TI - Comparison of tear film thickness in three species determined by the glass fibre method and confocal microscopy. AB - Using optical methods to measure tear film thickness we previously found that values were substantially larger than earlier estimates by mechanical and chemical techniques. The predominant component was found to be mucus, not aqueous fluid. This article describes the results of using confocal microscopy to re examine an earlier method of measuring thickness, the glass fibre technique. Confocal microscopy enabled visual identification of tear and epithelial surfaces and accurate measurement of the separation between them, giving the full film thickness. Consecutive measurements were made by each method in eyes of 22 animals of three species. Measurements using fibres in rabbits gave results of 7 microns, replicating earlier findings. However, the fibre technique gave the same result in all species. Values were not correlated with measurements by microscopy, which ranged from 10 to 14 microns. We propose that fibres were too flexible to be pressed through all mucus layers and contact epithelium, and as a result this method underestimated film thickness. PMID- 8224312 TI - Infective panophthalmitis simulating malignant melanoma of the choroid in a patient with myelodysplasia. PMID- 8224313 TI - Discrete metastasis to an extraocular muscle. PMID- 8224314 TI - Acute haemorrhagic conjunctivitis epidemic in the Dominican Republic. PMID- 8224315 TI - Dangers of direct injection of antimetabolites into filtration blebs. PMID- 8224316 TI - Visual fields and diabetic retinopathy. PMID- 8224317 TI - Guidelines for the safety assessment of microbial enzymes used in food. AB - The regulatory approach in the UK to the safety assessment of microbial enzymes has been reviewed. The Committee on the Toxicity of Chemicals in Food, Consumer Products and the Environment (COT) devised, in December 1992, a strategy based on toxicological testing of a representative batch of the final purified fermentation product and achieving a high standard of quality control through the development and maintenance of quality assurance specifications. PMID- 8224318 TI - Comparison of two clean-up methodologies for the gas chromatographic/mass spectrometric determination of low nanogram/gram levels of polynuclear aromatic hydrocarbons in seafood. AB - The March 1989 oil spill in Alaska prompted the Food and Drug Administration (FDA) to conduct a thorough investigation of clean-up methodologies aimed at determining low ng/g (ppb) levels of polynuclear aromatic hydrocarbons (PAHs) in seafood. The clean-ups from a modified FDA method and a National Marine Fisheries Service (NMFS) method were evaluated on the basis of the determination of 18 PAHs at levels ranging from 1 to 5 ppb by gas chromatography/mass spectrometry. In the modified FDA method, seafood extracts were purified by a liquid-liquid partition followed by a three-step elution through silica, alumina, and C18 solid-phase extraction cartridges. In the NMFS method, seafood extracts were purified by column chromatography through a deactivated silica gel/alumina column and a gel permeation high performance liquid chromatography column. Both methods quantitated 18 PAHs at levels ranging from 1 to 5 ppb. With the exception of naphthalene, average recoveries based on internal deuterated standards ranged from 73 to 144% for the modified FDA method and 63 to 106% for the NMFS method. PMID- 8224319 TI - Polycyclic aromatic hydrocarbons in smoked food products and commercial liquid smoke flavourings. AB - Smoked foods including turkey, pork, chicken, beef and fish products were screened for the presence of carcinogenic and non-carcinogenic polycyclic aromatic hydrocarbons (PAHs). Eighteen commercial liquid smoke flavourings and seasonings were also analysed. Total PAH concentrations in smoked meat products ranged from 2.6 micrograms/kg in a cooked ham sample to 29.8 micrograms/kg in grilled pork chops, while those in fish products ranged from 9.3 micrograms/kg in smoked shrimp to 86.6 micrograms/kg in smoked salmon. Total concentrations of the carcinogenic PAHs (benzo[a]anthracene, benzo[b]fluoranthene, benzo[a]pyrene, dibenzo[a,h]anthracene, and indeno[1,2,3-c,d]pyrene) ranged from non-detectable in several meat products to 7.4 micrograms/kg in grilled pork chops, and from 0.2 micrograms/kg in trout to 16.0 micrograms/kg in salmon. In liquid smoke flavourings and seasonings, total PAH concentrations ranged from 6.3 to 43.7 micrograms/kg, with the carcinogenic PAHs ranging from 0.3 to 10.2 micrograms/kg. PMID- 8224320 TI - Migration from plasticized films into foods. 6. Hydrolysis of polymeric plasticizers under simulated gastric and intestinal conditions. AB - The polymeric plasticizers poly(butylene adipate) and poly(propylene adipate), and a low molecular weight oligomer fraction from each, were exposed in vitro to simulated gastric and intestinal hydrolysis in order to study their likely fate after ingestion of foods contaminated by migration from plastics packaging. Analysis for loss of the parent compounds indicated that partial hydrolysis occurred under simulated gastric conditions, with the low molecular weight oligomers being the most susceptible. Analysis for one of the starting materials, adipic acid, revealed however that the hydrolysis did not result in a significant conversion to free monomer under either simulated gastric or intestinal conditions. This is explained on the basis of partial hydrolysis of the plasticizers to smaller oligomer units but with the more resistant ester linkages resisting complete breakdown to monomer. PMID- 8224321 TI - Speciation of organotins in poly(vinyl chloride) products. AB - Extraction studies on butyl- and octyltins were conducted with three poly(vinyl chloride) (PVC) product types: clear food container, rigid pipe and flexible membrane. Three solvents, tetrahydrofuran, xylene and methylene chloride were evaluated for extraction efficiency. Methylene chloride extracted more than 97% of the total extractable organotin in two extractions and resulted in the highest recoveries of analytes from all three PVC products. Method detection limits ranged from 0.3 to 0.9 microgram alkyltin/g PVC. The mean precision of the method is 6.8% relative standard deviation. Food industry PVC product samples contained between < 0.8 and 8751 micrograms/g octyltins and < 0.3 and 4.7 micrograms/g butyltins. Butyltins (< 0.9-5985 micrograms/g) were detected in potable water pipe samples. Pipe used in industrial applications contained both butyl- (13-1501 micrograms/g) and octyltins (701-3033 micrograms/g). PMID- 8224322 TI - Analysis of migrants from nylon 6 packaging films into boiling water. AB - Ultra-violet spectrophotometry (UV), high performance liquid chromatography (HPLC) and liquid chromatography coupled to mass spectroscopy (LC-MS) were used to identify and quantify oligomers extracted with boiling water from two different nylon 6 films used in boil-in-bag food packaging. The results indicated the loss of up to 1.5% of the original nylon film weight, into the boiling water, as caprolactam and cyclic oligomers up to the nonamer. Extraction time, thickness and type of film used, were found to be parameters which affected the levels of these migrants. These results will be relevant to situations in which food is cooked in the water used to heat the pouch contents. PMID- 8224323 TI - Overall migration and specific migration of bisphenol A diglycidyl ether monomer and m-xylylenediamine hardener from an optimized epoxy-amine formulation into water-based food simulants. AB - The overall and specific migrations of BADGE n = 0 monomer and m-XDA hardener from a BEPOX LAB 889 (Gairesa internal code), epoxy system cured at room temperature, into three water-based food simulants are studied. Hydrolysis of BADGE n = 0 was observed in all of these simulants, giving more polar products. We thus propose changing the EEC Directives, which at present only legislate for levels of BADGE n = 0 monomer in the simulants, to include the hydrolysis products of BADGE monomers. Another alternative would be to express all the migration levels due to BADGE and its derived products in terms of BADGE itself. PMID- 8224324 TI - Migration of styrene monomer from thermoset polyester cookware into foods during high temperature applications. AB - Migration of styrene from thermoset polyester cookware into foods has been studied during normal cooking applications and for the fatty food simulant olive oil under high temperature test conditions. The analysis of foods was by automated headspace gas chromatography/mass spectrometry (GC/MS) in the selected ion mode. Cyclohexane was added to the slurried foods to improve headspace partition and deuterated-styrene was the internal standard. To achieve adequate sensitivity with olive oil, styrene was extracted and concentrated by a steam distillation method and then determined by GC/MS. Styrene in foods cooked in thermoset polyester articles was in the range < 5 to 5 micrograms/kg and 5 to 30 micrograms/kg where the polyester contained 9 and 380 mg/kg residual monomer respectively. Testing for 2 h at 175 degrees C into olive oil resulted in significantly higher migration of styrene than seen for other foods, although there was a marked decrease in migration on repeat-use. Testing of the complete articles as sealed systems filled with olive oil led to higher migration than testing by total immersion of plaques. Of the test methods used with olive oil, testing as filled articles with no lidding gave migration results closest to (but generally higher than) those seen for real foods. PMID- 8224325 TI - Survey of imported green coffee beans for pesticide residues. AB - The US Food and Drug Administration carries out incidence/level monitoring in order to acquire data on the presence and amounts of pesticide residues in particular commodity/chemical combinations. In the survey reported here, imported green coffee beans were analysed for a variety of pesticide chemicals. A total of 60 green coffee samples were collected from 21 countries that are major exporters of coffee to the United States. The samples were analysed for organochlorine/organophosphorus, N-methyl carbamate, benomyl group and EBDC residues. Four samples had detectable residues: chlorpyrifos, 0.01, 0.02 and 0.04 ppm and pirimiphos-methyl, 0.01 ppm. The majority (93%) of the green coffee samples analysed in this survey had no detectable pesticide residues. PMID- 8224326 TI - Determination of ivermectin residues in the tissues of Atlantic salmon (Salmo salar) using HPLC with fluorescence detection. AB - A sensitive, accurate and reproducible assay for the determination of residual concentrations of ivermectin in the tissues of farmed Atlantic salmon is described. The assay employs the clean-up of an acetonitrile extract of salmon tissue on a Bond-Elut C8 column, followed by derivatization with trifluoroacetic acid anhydride. Following separation using HPLC, 22,23-dihydroavermectin B1a is determined using fluorescence detection. The limit of detection of the assay is 1 ng per g tissue. Liver, taken from fish treated experimentally with ivermectin, was found to contain higher concentrations of 22,23-dihydroavermectin B1a than any other tissue examined. The half-life of 22,23-dihydroavermectin B1a was similar in all tissues studied, ranging from 89 to 98 degree days. PMID- 8224327 TI - A survey of ethyl carbamate in beverages, bread and acidified milks sold in Denmark. AB - A sensitive gas-chromatographic method of analysis using mass specific detection has been applied to a study of ethyl carbamate in the Danish diet. Beverages and foods which were assumed to give a major contribution to the exposure of the Danish population to this potentially carcinogenic compound were investigated. Average values, ranges obtained for concentrations of ethyl carbamate and the number of analyses were 534 micrograms/l (< 5-5000 micrograms/l, n = 22) in spirits, 30 micrograms/l (7-61 micrograms/l, n = 14) in fortified wines, 7 micrograms/l (< 3-29 micrograms/l, n = 57) in wine, 3 micrograms/l (0.2-6.6 micrograms/l, n = 50) in beer, 3.5 micrograms/kg (0.8-12 micrograms/kg, n = 33) in bread and 0.2 microgram/kg (< 0.1-0.3 microgram/kg, n = 19) in yogurts and other acidified milk products. It is estimated that the average daily intake of ethyl carbamate is approximately 2 micrograms per person. The 5% of Danish males who have the highest alcohol intake probably consume more than 7 micrograms of ethyl carbamate per day. PMID- 8224328 TI - Determination of histamine in fish using an enzymic method. AB - The present paper describes a quick and simple enzymic method for evaluating histamine content in fish. After preparing a perchloric acid extract, it was neutralized to pH 6.8 with KOH. The action of a diamine oxidase on the histamine caused the formation of hydrogen peroxide. The addition of a second enzyme, a peroxidase, in the presence of hydrogen peroxide and a chromogen (leuco crystal violet) in reduced form (colourless) facilitated its oxidation into crystal violet (coloured). Following a two-hour incubation period, absorbance was measured at 596 nm. The correlation between histamine level and absorbance was acceptable in the concentration range from 3 to 30 mg/kg of histamine (r = 0.9946; p < 0.001). The accuracy of the method, expressed by CV, varied between 2.6% and 4.9%, and the recovery between 95.7% and 100.1%, depending on the concentration of analyte in the sample. The diamine oxidase used was very selective in relation to the histamine. Only the presence of tyramine, when histamine concentration was low (< 10 mg/kg), tended to interfere to a slight degree in the technique. Using a regression analysis, no statistically significant differences were found between the results obtained from the analysis of 18 tuna fish samples by HPLC and the enzymic method (r = 0.9867; p < 0.001). PMID- 8224329 TI - The use of Daphnia magna for detection of okadaic acid in mussel extracts. AB - Okadaic acid (OA), the main toxin responsible for diarrheic shellfish poisoning (DSP) has toxicity for Daphnia magna (EC50 = 15 +/- 1.8 micrograms/l). A Daphnia bioassay was developed and used to analyse okadaic acid in mussel extracts. A linear correlation was found between OA concentration determined by the Daphnia bioassay and by HPLC assay (r = 0.85; p < 0.001). The Daphnia bioassay can measure OA levels 10 times below the threshold of the mouse bioassay method. It is an inexpensive sensitive tool which can be used in replacement of mouse bioassay for the screening of OA and some co-extracting toxins in mussel extracts. PMID- 8224330 TI - Imaging using solid-state detectors. AB - Whether they know it or not, many dentists already have adapted solid-state imaging devices into their offices in the form of an intraoral or video-camera system for esthetic dentistry and patient education. These dentists own charge coupled devices. The technology, however, had been accepted well before instruction in the imaging principles was available. With the addition of digital radiographic systems, the dentist who is "buying into" these contemporary imaging devices must become familiar with the imaging principles behind them, and with the applications of these systems to dental problems or tasks. This article attempts to bring the dentist "up to speed" with this technology and discusses its present and future applications. In this way, the clinician will move past the simple "show-and-tell" stage or patient education and begin to see the true potential of imaging for more accurate detection of disease, for quantitating changes in that disease through time, and for making better clinical decisions about patient treatment. PMID- 8224331 TI - Digital panoramic and extraoral imaging. AB - Intraoral, panoramic, and extraoral radiographs are the primary means of diagnosing hard-tissue disorders of the dentomaxillofacial region. These imaging methods require the use of x-ray film and subsequent chemical processing to produce diagnostic images. A goal of recent research has been the replacement of this film-based technology with computer-based devices that use electronic or storage phosphor receptors to record the x-ray image in a digital format. This article discusses some of these emerging technologies and their potential effect on the future of panoramic and extraoral radiology. PMID- 8224332 TI - Digital subtraction radiography. AB - Subtraction radiography offers greater visualization of radiographic changes between a pair of radiographs by subtracting out the unchanging background distractions. Originally, subtraction was achieved by using positive and negative prints in angiography. The advent of digital imaging devices has led to validation of quantitative measurements from digital subtraction images. As well, computer software manipulations have allowed for more flexibility in the clinical application of subtraction imaging. The clinical application of digital subtraction radiography is discussed along with clinical cases. PMID- 8224333 TI - Tomography. AB - Conventional tomography is used in dentistry to demonstrate objects lying in a plane of interest distinct from their surrounding anatomic structures. Through technique parameters, which generate the controlled blurring of structures above and below the plane of interest, anatomic structures are visualized without the superimposition noted on other types of plain radiographs. In this article, the basic principles of tomography have been reviewed and the clinical applications for conventional tomography in dentistry have been described. PMID- 8224334 TI - Computed tomography. AB - Although CT scanning has been available for less than 20 years, it has made a major impact on the practice of dentistry, particularly in oral and maxillofacial surgery, in the diagnosis and management of a wide variety of oral lesions. The use of computers to reconstruct x-ray attenuation data into an image allows the clinician to view pathology from many different perspectives, unobstructed by other anatomic structures, and to adjust contrast to concentrate on the soft tissue and hard-tissue pathology in turn. Advances in computer software already allow three-dimensional visualization of pathology and anatomy, but additional refinements in this technology still are expected. PMID- 8224335 TI - Magnetic resonance imaging of the head and neck. AB - MR imaging of the head and neck region has become the modality of choice for evaluating both malignant and benign tumors, functional derangements such as in temporomandibular joint dysfunction, and many congenital lesions. With improved scanner design and with newer pulse sequences becoming available, one can expect increased use of MR imaging in the future. PMID- 8224336 TI - Two-dimensional-three-dimensional reconstruction computed tomography techniques. AB - A brief but comprehensive review of the technologic basis of computed tomography leading to the generation of more specialized and sophisticated reformatted and reconstructed two- and three-dimensional images is presented. Elements that influence the quality of such images are emphasized. Examples of clinical applications in maxillofacial and dental practice are illustrated. PMID- 8224337 TI - Temporomandibular joint imaging. AB - New advances in imaging have added a significant amount of information, which has resulted in more accurate and detailed diagnosis of temporomandibular joint disorders. In the future, treatment methods may be devised that can use effectively this diagnostic information. PMID- 8224338 TI - Implant radiology. AB - Implant procedures are becoming commonplace in the dental office. Root form implants are placed by the general dentist, and no longer remain in the realm of dental specialists. Pre- and postsurgical radiographic evaluation of endosseous or root form implants is central to the success of the procedure. This article reviews the applications and limitations of available imaging modalities--from intraoral radiographs to computed tomography imaging--to help the clinician decide which imaging technique is best suited to the task of radiographic evaluation of implants and implant sites. PMID- 8224339 TI - Teleradiology for dentistry. AB - As the world enters the "telecosm," the importance of rapid information transfer will grow. It is not unusual for the average American to have a beeper on his or her belt, a cellular telephone in the car, a telephone with remote access answering machine, voice mail, call forwarding, a home fax, and a personal computer networked with a commercial purchasing and information retrieval service such as CompuServe or Prodigy. Conventional cellular telephones, introduced in 1983, numbered approximately 6.4 million in the United States by January, 1992. This is a measure of the importance placed on rapid interpersonal communication. Undoubtedly, patients will begin to expect at least the same degree of efficiency in communications on the part of health care providers, including dentists. There are great potentials for expediting prior approval and reducing costs for dental care as dentists obtain new, lower-priced, direct digital imaging systems, especially as ISDN and electronic mail systems become more available and less costly. It should be possible to reduce insurance costs by minimizing the numbers of clerical staff needed to open, check, and return submitted radiographs. Centralized diagnostic centers for second or expert opinions both for the dental profession and for the insurance carriers are additional pluses. There are potential savings in mailing costs and in reducing travel required of the patient living in a remote region to obtain a second opinion. Transmission using commercially available access to Internet/NSFNET is already possible for a small monthly fee. Nothing, other than a basic personal computer with graphics capabilities, is needed to transmit and receive files once the image is in a digital form. Electronic imaging and image transmission are here. PMID- 8224340 TI - Computer-assisted interpretation in radiographic diagnosis. AB - The application of computer-aided procedures to analyze digital radiographs will help to increase the quality of radiographic diagnosis. Image processing is applied to improve image quality and to extract features from digital radiographs. Computer programs can be used to integrate image data and patient data originating from different sources. The aim of these procedures is to support the clinician in the decision-making process. PMID- 8224341 TI - The oral surgery/orthodontic interface: 2. Local causes of malocclusion. AB - This is the second of two articles discussing the minor oral surgery/orthodontic interface. The vital role played by the general dental practitioner in monitoring the normal development of the occlusion and the early diagnosis of the local causes of malocclusion was emphasized in the first article. Part 2 discusses the problems associated with the prominent labial frenum, supernumerary teeth and submerged deciduous molars. PMID- 8224342 TI - Hydrochloric acid/pumice microabrasion technique for the removal of enamel pigmentation. AB - Products for lightening teeth are appearing on the market with ever-increasing frequency. Heavy advertising, coupled with heightened public awareness and expectations of an aesthetically pleasing smile, have resulted in increased patient demand for treatment to improve all types of tooth discoloration. Although it is the role of the dental profession to provide the services demanded by the consumer, it is also our duty to be discerning and to re-assess continually the techniques we use, for both efficacy and safety. To do this, long term follow up of clinical techniques is essential. This paper contains the clinical results of teeth treated over 4.5 years using the hydrochloric acid/pumice microabrasion technique to remove enamel opacities and pigmentation. The subject under discussion is that of enamel discoloration: intrinsic staining with its origins in dentine, such as that caused by the ingestion of tetracycline antibiotics during odontogenesis, is not amenable to this form of surface treatment. PMID- 8224343 TI - Resin luting materials: the current status. AB - Compared with composite resin restorative materials, which have evolved considerably since their introduction over two decades ago, the development of resin-based luting materials has been slow. Only with the introduction of restorative techniques such as resin-bonded bridgework, porcelain veneers, and composite resin or ceramic inlays, have resin-based luting materials gained importance. PMID- 8224344 TI - Orthodontic treatment and the wind instrument. AB - Many children are learning to play musical instruments at the same time as they are undergoing orthodontic treatment. If a patient or parent enquires about how orthodontic treatment might affect their child's playing, what advice could you give them? PMID- 8224345 TI - Antiseptic mouthwashes in modern dental practice. AB - This article is based on the winning entry of the Oraldene Student Essay Competition which was announced in Dental Update in the November 1992 issue. On the opposite page Miss Ahmed can be seen accepting her prize of 500 pounds from Professor C.E. Renson and Professor F.C. Smales. PMID- 8224346 TI - Management of a tooth affected by idiopathic coronal resorption. AB - This article reports on the restoration of a permanent mandibular canine affected by significant idiopathic coronal resorption. A particular restorative approach and the alternative treatment options are discussed. The need for careful examination of diagnostic radiographs is emphasized. PMID- 8224347 TI - Oral cancer: new insights into pathogenesis. AB - Understanding the pathogenesis of cancer may well lead to improved diagnostic tests, preventive treatment and management. This paper reviews the gene changes, the evidence implicating viruses in the aetiology of oral cancer, and the mechanisms whereby viruses may affect gene function and thereby contribute to carcinogenesis. PMID- 8224348 TI - Festschrift in honour of Claude Rimington"s 90th birthday. PMID- 8224349 TI - Was Hippocrates the first to describe a case of acute porphyria? PMID- 8224350 TI - Biochemistry of porphyria. AB - 1. The porphyrias are a group of metabolic disorders arising from defects in the haem biosynthetic pathway. Most forms are inherited as Mendelian autosomal dominants, but some types are recessive and others acquired through exposure to porphyrinogenic drugs and chemicals. There is a linked group of diseases, which are not porphyrias, but have in common alterations of haem biosynthesis. 2. The processes of haem biosynthesis are now well understood and the molecular biology of the functions and dysfunctions in the porphyrias are currently an area of intensive investigation. 3. The acute porphyrias, Acute Intermittent Porphyria, Variegate Porphyria and Hereditary Coproporphyria are of most importance since attacks of these may be life-threatening. 4. These diseases that usually present with a neurovisceral attack are characterized by excess production of the porphyrin precursors, 5-aminolaevulinate and porphobilinogen because of lowered activity of Porphobilinogen deaminase. 5. A variety of factors may precipitate these attacks including various drugs, alcohol, smoking, dieting or fasting and variations in steroid hormone levels. 6. The non-acute porphyrias are largely dermatological conditions, which present clinically as cutaneous photosensitivity. The dermatological changes are caused by the photosensitizing properties of circulating porphyrins and are accompanied by systemic effects of these porphyrins. PMID- 8224351 TI - The role of lipoproteins in the delivery of tumour-targeting photosensitizers. AB - 1. Serum lipoproteins play an important role in the in vivo transport of several porphyrinoid derivatives having a moderate or high degree of hydrophobicity. 2. There appears to exist a correlation between the extent of photosensitizer association with low-density lipoproteins (LDL) and the efficiency of tumour targeting by some classes of photosensitizers, such as differently sulphonated porphyrins and phthalocyanines, haematoporphyrin dialkylethers and unsubstituted phthalocyanines and naphthalocyanines. 3. In all cases, LDL-carried photosensitizers are preferentially released to malignant cells; hence, direct cell damage appears to be the major determinant of tumour damage consequent to photodynamic therapy. 4. Present evidence suggests that the LDL-associated photosensitizer is accumulated by tumour cells largely via a receptor-mediated endocytotic process. 5. Thus, the use of delivery systems for orientating a systemically injected photosensitizer towards lipoproteins has been explored; promising results have been obtained by incorporation of the dye into liposomal vesicles, oil emulsions or inclusion complexes, as well as by precomplexation of the dye with LDL. 6. Moreover, a suitable choice of the chemical constituents of the delivery system and the experimental conditions allows one to modulate the photosensitizer distribution among the different lipoproteins. 7. The occurrence of tumour-targeting strategies other than the LDL pathway is briefly discussed. PMID- 8224352 TI - Biodistribution of photosensitizing agents. AB - 1. The features of neoplasia which predict for drug responsiveness are rapid growth and/or inefficient repair of damage, especially to DNA. 2. PDT has the advantage of yielding responses regardless of the growth fraction of a tumor, and repair appears to play only a minor role. 3. While an entirely different spectrum of tumors can be targeted with PDT, the perhaps unavoidable accompaniment is that a new set of rules for efficacy will need to be established. 4. The selectivity of PDT is based on the need for irradiation which can be directed, along with the short tissue half-life of the cytotoxic product, singlet oxygen. Sensitizers which target specific cellular organelles could promote PDT efficacy, if in vitro data (Woodburn et al., 1992b Photochem. Photobiol. 55, 697-704) can be translated into clinical practice. 5. It remains to be established whether total drug distribution to neoplastic tissues or concentration in specific sub-cellular sites is the more important factor. 6. Questions relating to the role of biodistribution as a factor in efficacy of PDT sensitizers of photosensitizers remain to be explored. Just as the political cartographers are grappling with changes in territorial boundaries of known lands, we continue to clarify the rules relating to PDT boundaries. In this regard, it is clearly important for determinants of pharmacokinetics and biodistribution to be evaluated and understood. 7. Once clinical reports on the "second generation" agents are published, we may get a better picture, although it is not unusual for clinical reports to raise more questions than they answer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224353 TI - The biochemical approach to cancer therapy: a short essay. PMID- 8224354 TI - Photodynamic action of endogenously synthesized porphyrins from aminolevulinic acid, using a new model for assaying the effectiveness of tumoral cell killing. AB - 1. The effectiveness of the photodynamic action of porphyrins, was studied by means of the tissue explant culture technique. A murine tumor tissue explant was incubated in a medium containing 0.6 mM of ALA for periods of 1 and 2 hr; total porphyrins synthesized under these conditions were of the same level as those found in our previous in vivo experiments. The explants were then irradiated for 30 min with He-Ne laser of 3.5 mW output power placed at a distance of 10 cm. Controls of non-irradiated tumor tissue slices incubated with and without ALA were performed. Immediately after irradiation, inocula of exactly 1 mm3 of the irradiated and non-irradiated tissue were subcutaneously injected under the right and left flanks of the same animal, respectively. The growth of the tumor was measured 15, 20 and 25 days after implantation. 2. Results obtained showed that the explants that were incubated for 1 hr with ALA and irradiated, reaching a concentration of 2.8 micrograms porphyrins/g tissue, produced a reduction of 50 70% of tumor size as compared with the non-irradiated controls incubated with ALA. Explants incubated for 2 hr, reaching a concentration of 4.6 micrograms porphyrins/g tissue, produced from 60% to complete lack of tumor growth. The effectiveness index (EI) of photoirradiation was calculated on the basis of the tumor growth in irradiated and non-irradiated tumors. EI was nearly 100% showing almost complete tumor cell destruction for tumor irradiated for 2 hr with 0.6 mM ALA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224355 TI - Collapse of K+ and ionic balance during photodynamic inactivation of leukemic cells, erythrocytes and Staphylococcus aureus. AB - 1. The immediate and fast ionic fluxes in Friend erythroleukemic cells (FELC), erythrocytes and Staphylococcus aureus during short intervals of porphyrin mediated photosensitization were determined uniquely by X-ray microanalysis (XRMA) combined with electron microscopy. 2. Photodynamic inactivation of FELC was mediated by either endogenous protoporphyrin induced by 5-amino levulinic acid (5-ALA), or Photofrin-II. We describe the predominant phenomena of > 85% K loss within 2-10 min of photoactivation. However the accompanied Na inflow and the collapse of the cellular balance of elemental-composition were inconsistent and acted as a function of cell damage. 3. Erythrocytes treated with hematoporphyrin (HP) lost most of their intracellular K yet instantly gained Na. Nevertheless the K/Na molar ratio of the control erythrocytes was nearly 12/1 while after photosensitization and K loss it changed to 1/1. 4. The S. aureus bacteria photosensitized with HP showed entire K-loss as well as marked Na efflux which increased with irradiation time; this was accompanied by the decline of other cell elements. 5. The prevailing K loss in FELC, erythrocytes and bacteria during the first minutes of photosensitization is deduced to be an immediate primary consequence of the photodynamic effect, while other ionic changes are joined in order with the development of cellular damage. PMID- 8224356 TI - Kinetics of porphyrin accumulation in cultured epithelial cells exposed to ALA. AB - 1. The kinetics of porphyrin accumulation in cultured mammalian epithelial cells (CNCM-I-221) during exposure to ALA was investigated. 2. The total porphyrin synthesized is a function of ALA concentration and the incubation time. The cellular porphyrin content exhibited a saturation pattern, reaching a plateau at about 0.04 fmol porphyrins/cell. A biphasic time-dependent increase in the total porphyrin synthesized was observed. 3. After 3 hr of exposure to ALA the rate of synthesis increased to almost twice the initial rate, reaching between 0.02 and 0.05 fmol porphyrins/cell/hr depending on serum concentration in the medium. 4. Two effects of FBS on ALA-stimulated porphyrin accumulation were observed. Greater total porphyrin synthesis was found when incubations were made in 10% FBS compared to those in 1% FBS. 5. The higher serum concentration also caused a greater release into the medium of the porphyrins generated in the cells with a calculated half-life of 24 min in 10% serum-supplemented medium compared with 62 min in 1% serum. 6. The results obtained from cell synchronization experiments suggest that there is little obvious cell cycle-dependent variation in the synthesis of porphyrins from ALA. 7. The small differences in the intracellular porphyrin content that were observed may be attributed to a slight reduction in the rate of loss of porphyrins in G2/M cells. PMID- 8224357 TI - A compendium of reviews in biochemistry and molecular biology published in the first half of 1993. AB - 1. A compendium of reviews and mini-reviews in Biochemistry and Molecular Biology published in the first half of 1993 is presented. In all 554 titles are listed from 103 different publications. 2. This compendium presents the references by Journal Name. Keywords have been included with each reference to increase the value of the collection. Keyword and author cross-reference indexes are not included but are available in the electronic database from which this version was constructed. Should anyone wish to have this information in electronic form it can be distributed on MS-DOS formatted floppy disks in either Reference Manager or Medline format. The author should be contacted for details of the number of preformatted floppy disks required. PMID- 8224358 TI - The flow theory of enzyme kinetics: role of solid geometry in the control of reaction velocity in live animals. AB - 1. When blood flows, membranes are bombarded with ions etc., whose entry creates an ATP demand proportional to flow rate. Also proportional to flow rate is ATP production from oxidation of substrates [S] from the same blood volume. 2. O2 is limiting and reaction velocity at rest (metabolic rate) is determined by flow rate, F, but not by [S]. 3. Since resting blood O2 A-V difference is about 5 vol%, 11 circulated produces about 0.25 kcal in mammals, birds or warm reptiles. 4. Where O2 is not limiting, as in most amino acid deaminations, V = K F[S] with K a constant unrelated to Km. 5. At equal blood vol/kg, solid geometry dictates that the average cross-sectional area of major vessels/kg will be an inverse function of body mass. The smaller the animal, the shorter the vessels, the "thicker" the vessels/kg body wt, and at any one blood pressure, the higher the flow/kg/hr. If a man's major vessels were equal in cross-section/kg to those of a shrew, it would take 224 l of blood to fill them. 6. Growth decreases flow/kg (and therefore metabolic rate), by decreasing vessel cross-section/kg without changing blood pressure or linear velocity of flow. 7. Surface area/g, body wt to some power, average vessel length/kg, circulation time and average major vessel cross-sectional area are all related mathematically. PMID- 8224359 TI - DNA breakage and inactivation resulting from hydroxylamine and/or cis diamminedichloroplatinum(II) interactions with plasmid DNA. AB - 1. Occurrence of lesions induced in plasmid DNA by cis-DDP and by HA was quantified both as a transforming activity and as conformation integrity of supercoiled pBR322 DNA. Fifty per cent decrease of the biological activity of plasmid DNA, not accompanied by measurable change of DNA conformation, was observed after a single exposure of DNA to cis-DDP (1 hr/37 degrees C). 2. HA induced conversion of supercoiled DNA to other topological forms in a dose dependent manner. 3. One- and two-strand DNA breaks were determined electrophoretically with high sensitivity. Cis-DDP exposed DNA relaxed at 30 times lower HA concentration compared to intact DNA. 4. This effect may be connected with a local distortion of DNA structure at the cis-DDP--DNA bond, which makes possible high effectivity of HA-DNA interaction. 5. On other hand, biological activity stayed at the 50% level despite breaks induced in DNA. 6. This finding supports the idea that DNA breaks occur at the locations which were modified during the exposure of DNA to cis-DDP. 7. The importance of the DNA structure during interaction with HA may be seen during HA-DNA interaction at heat-denaturation of supercoiled DNA. At this condition, the DNA breaks were induced at 100 times lower concentration of HA. 8. We conclude, on the basis of these results published earlier, that local distortion of supercoiled DNA structure, which is caused by the cis-DDP bond, and the local DNA uncoiling caused by heat-denaturation are related to high HA-DNA reactivity. PMID- 8224360 TI - Purification and characterization of lipoprotein lipase from the white adipose, skeletal muscle, cardiac muscle, mammary gland and lung tissues of the rat. AB - 1. Lipoprotein lipase (LPL) was isolated from five rat tissues: white adipose, skeletal muscle, cardiac muscle, mammary gland and lung. 2. Specific activity of the preparations varied from 75 U/mg for skeletal muscle and 720 U/mg for adipose. 3. The preparations were further analysed using SDS-PAGE and a single component identified. The mol. wt of 61,000 Da of this component was consistent for all five of the tissue sources. 4. Significant differences in the values of the isoelectric points of the enzyme species were revealed. The values varied from 7.23 (SEM 0.022) for cardiac and lung to 7.51 (SEM 0.037) for mammary. 5. Two-dimensional electrophoresis, using isoelectric focusing in the first dimension and SDS-PAGE in the second revealed differences in the patterns of stained material derived from the five tissue sources. PMID- 8224361 TI - Regulation of spermidine transport in L1210 cells. AB - 1. 1 mM 2-amino isobutyric acid (AIB), glutamine or asparagine when preincubated for 3 hr with L1210 cells promoted a marked increase in the rate of spermidine uptake. 2. Cycloheximide also increased the transport rate and completely prevented the increase due to AIB. 3. Trifluoperazine and iso-H7 inhibited the uptake of spermidine, much less the uptake of AIB. 4. Adenosine promoted an increase in the uptake of AIB, a decrease in that of spermidine. 5. Hypotonic stress also increased the rate of spermidine transport. This modification was only partially prevented by cycloheximide. 6. Okadaic acid had no effect on this increase, whereas it prevented the increase of ODC activity. PMID- 8224362 TI - Biochemical investigation of leukocyte functions during lithium therapy. AB - 1. The effect of lithium on phagocytic activity of polymorphonuclear leucocytes (PMNL) has been investigated by measurements of glucose-6-phosphate dehydrogenase (G6PD), NADPH oxidase and myeloperoxidase (MPO) both in lithium treated rats and lithium treated infected rats. 2. The results have been compared with two control groups, one of which was without lithium treatment and the other was only infected. 3. In the first experimental group increased activities of these enzymes have been observed, while in lithium-treated infected rats there was a decrease in the activities of the same three enzymes. 4. It is proposed that defense mechanisms against infection fail during the lithium treatment. PMID- 8224363 TI - Cloning and sequence analysis of A cDNA encoding bovine cytosolic aspartate aminotransferase. AB - 1. Complementary DNA encoding cytosolic aspartate aminotransferase was isolated from an adult bovine heart library. 2. The amino acid sequence deduced for the protein (412 amino acids) is extremely similar (> 94% identity) to that of porcine cytosolic aspartate aminotransferase but interesting differences were noticed comparing the position of cysteine residues. PMID- 8224364 TI - Differences in inducibility of cytochrome P-4501A1, monooxygenases and glutathione S-transferase in cutaneous and extracutaneous tissues after topical and parenteral administration of beta-naphthoflavone to rats. AB - 1. The effect of topical and parenteral administration of beta-NF to rats on cytochrome P-4501A1, monooxygenases and glutathione S-transferase enzyme activities in various tissues were studied. 2. Topical and I.P. administration of beta-NF to rats resulted in variable induction responses (2-28-fold) in various tissues. Benzo(a)pyrene hydroxylase, ethoxyresorufin-O-deethylase, ethoxycoumarin O-deethylase, and GST activities in liver, kidney and skin were induced to higher extent than in other tissues following both topical and i.p. administration. 3. Western blot analysis of microsomal preparation from beta-NF-treated rats using monoclonal antibody to P-4501A1 showed a marked induction of P-4501A1 enzyme protein in skin and liver. 4. These results suggest that both topically as well as parenterally administered xenobiotics may influence drug metabolizing capacity of skin and other tissues. PMID- 8224365 TI - Structure of heparan sulfate from the fresh water mollusc Anomantidae sp: sequencing of its disaccharide units. AB - 1. The disaccharide sequences of a heparan sulfate isolated from Anomantidae sp. was determined with the aid of heparitinase I, heparitinase II from Flavobacterium heparinum, mollusc beta-glucuronidase and alpha-N acetylglucosaminidase besides nitrous acid degradation and chemical analyses. 2. Like the mammalian heparan sulfates the mollusc heparan sulfate is composed of different oligosaccharide blocks of N-acetylated disaccharides, N-sulfated disaccharides and N,6-sulfated disaccharides and has in its nonreducing end the monosaccharide glucosamine 2,6-disulfate. 3. The oligosaccharides produced by heparitinase I degradation contain at their reducing ends a N-acetylated, 6 sulfated disaccharide. 4. These and other results lead to the conclusion that the general structure of the heparan sulfate is maintained through evolution. PMID- 8224366 TI - Stimulation of plasmin activity in cultured human fibroblast cells by Porphyromonas endodontalis. AB - 1. Plasmin activity in the conditioned medium of Gin-1 cells, a human gingival fibroblast cell line, was stimulated by Porphyromonas endodontalis, a putative pathogen of oral submucous abscesses, in a time- and dose-dependent manner. 2. P. endodontalis stimulated the activity of plasminogen activator in both the conditioned medium and the cell lysate. The plasminogen activator in Gin-1 cells was approx. 50 kDa by zymography. 3. The conditioned medium of Gin-1 cells exposed to P. endodontalis stimulated the conversion of human serum prekallikrein to kallikrein. 4. These results suggested that P. endodontalis stimulates the plasminogen activator-plasmin system in Gin-1 cells, and that activated plasmin plays a role in the progress of periodontal tissue inflammation. PMID- 8224367 TI - Purification and characterization of carbonyl reductases from bovine liver cytosol and microsome. The cytosolic enzyme has a novel 3 alpha/17 beta hydroxysteroid dehydrogenase activity. AB - 1. Carbonyl reductase, which is distributed in both cytosolic and microsomal fractions in bovine liver, were purified to homogeneity on 12.5% sodium dodecylsulfate-polyacrylamide gel electrophoresis and shown to have molecular weights of 32 kDa and 68 kDa, respectively. 2. Both carbonyl reductases can catalyze the reduction of many carbonyl compounds including ketone, quinones and aldehyde with relatively low Km values. 3. From the absorption spectrum result, microsomal carbonyl reductase closely resembles cytochrome P-450 reductase. 4. Cytosolic carbonyl reductase is a novel enzyme which can act on both testosterone and androsterone at low concentration. PMID- 8224368 TI - In vivo effect of berenil on rat liver polyamine metabolism. AB - 1. Berenil, administered to rats in vivo, promoted a decrease in liver SAMDC activity, but an increase in ODC and SAT activity. 2. Its effect on ODC was completely prevented by cycloheximide, that on SAT only partially. 3. Berenil had no effect on ODC activity in adrenalectomized rats. Adrenergic antagonists counteracted the effect of Berenil on ODC activity. 4. Polyamine content was increased. The maximum modification was observed for putrescine and N1 acetylspermidine. PMID- 8224369 TI - Human urinary excretion of L-histidine-related compounds after ingestion of several meats and fish muscle. AB - 1. After oral administration of the muscle of skipjack tuna, about 90% of ingested anserine (Ans) was excreted quickly into urine as Ans (8%) and pi methylHis (82%), indicating the fast decomposition of Ans into pi-methylHis. This was also the case for chicken muscle ingestion. 2. After eel muscle ingestion, carnosine (Car) excretion was only 1% of the ingested whereas almost no increase was found in His, a constituent of Car, indicating the re-utilization of this essential amino acid. Similar results were also obtained from beef and pork ingestion. 3. In all cases, the urinary excretion of these compounds reached a maximum within 7 hr after ingestion and returned to the level of meat-free diet within 40 hr. PMID- 8224370 TI - Isolation and N-terminal sequence of two low molecular weight calcium-binding proteins from pig granulocytes. AB - 1. Two small, abundant calcium-binding proteins were isolated from pig granulocytes. They were named p7A and p7B. Relative molecular masses were approx. 32,000 for p7A and 13,000 for p7B, when obtained by Sephadex G-75 gel filtration, while it was 7000 for both proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). 2. N-terminal sequence analysis suggests that p7A is homologous to human and mouse MRP-8 and that p7B may be related to human and mouse MRP-14, though some properties of the latter--such as mobility on SDS-PAGE- were found to be different. In addition, p7A and p7B could be resolved under native conditions, contrasting with the fact that human and mouse MRP-8/MRP-14 form noncovalent complexes. PMID- 8224371 TI - Increased muscle glucose uptake in response to chronic glyburide treatment is not related to changes in glucose transporter (GLUT4) protein. AB - 1. The mechanism of action of glyburide (a sulfonylurea) on muscle has been investigated by measuring glucose uptake and glucose transporter (GLUT4) protein levels after chronic glyburide treatment. 2. A dietary induced insulin resistant rat model (4 wk of high-fat, high-sucrose feeding) was given glyburide (2 mg/kg/day) for 10 days and glucose uptake was measured in a perfused hindquarter preparation. 3. Protein levels of the GLUT4 glucose transporter were determined by Western analysis. 4. After 7 days of treatment, rats fed glyburide had lower blood glucose concentrations 2 hr (72 +/- 5 vs 103 +/- 12 mg/dl) and 24 hr (97 +/ 7 vs 123 +/- 7 mg/dl) after glyburide administration with no difference in serum insulin levels compared to vehicle treated animals. 5. Glucose uptake was approx doubled in basal state (0 insulin) in response to glyburide (2.8 +/- 0.4 vs 1.7 +/- 0.2 mumol/g per hr). 6. Maximal insulin (100 nM) stimulated glucose uptake tended to be higher in the glyburide treated group, but did not reach statistical significance (8.0 +/- 0.7 vs 7.0 +/- 0.6 mumol/g per hr). 7. Western analysis revealed no significant effect of glyburide on the GLUT4 protein level in skeletal muscle. 8. These results suggest that glyburide alters glucose uptake through some mechanism other than alterations in the level of the GLUT4 glucose transporter protein. PMID- 8224372 TI - Comparative studies of the Spi1 proteins of three equine alpha-1-proteinase inhibitor haplotypes following isolation by affinity chromatography. AB - 1. Antiproteinase deficiency can result in excessive proteinase-induced tissue damage. The major anti-elastase (Spi1) protein of equine alpha 1-proteinase inhibitor (alpha 1-PI) was isolated from the plasma/serum of three common haplotypes (I, L and U). 2. The N-terminal amino acid sequences of the three inhibitors were identical, but were only approx 65-77% homologous with two other published equine Spi1 sequences. 3. All three inhibitors complexed quickly and irreversibly with equine leucocyte proteinase 2A (kass = 2 x 10(7) M-1 sec-1). They were also efficient inhibitors of chymase (rat mast cell proteinase-II; kass = 2 x 10(5) M-1 sec-1; Ki = 2 x 10(-10) M). There was therefore no evidence of deficient inhibition in the Spi1 variants of the I,L and U haplotypes. PMID- 8224373 TI - Interactions and effects of 2-hydroxy-5-nitrobenzyl bromide on the bovine heart mitochondrial F1-ATPase. AB - 1. The F1-ATPase from bovine heart mitochondria was shown to chemically react and to absorb 2-hydroxy-5-nitrobenzyl bromide (HNB) with changes in catalytic properties. 2. The treatment of the enzyme with HNB at concentrations below 0.5 mM resulted in an increase of Vm and in an unchanged Km. Above 0.5 mM HNB elicited a concentration-dependent inhibition of F1. 3. HNB was found tightly bound to the enzyme epsilon-subunit whose tryptophan residue resulted modified. 4. The F1 activation appears the consequence of the covalent binding of the reagent to the enzyme, whilst inhibition results from non-covalent, reversible binding. 5. The possibility that the epsilon-subunit of mitochondrial F1-ATPase may influence the functional or regulating domain of the enzyme is discussed. PMID- 8224374 TI - The effect of tryptophan on dopa-oxidation by melanosomal tyrosinase. AB - 1. Tryptophan has been shown to inhibit dopa-oxidation by melanosomal tyrosinase. 2. The inhibition is of mixed-type with Ki = 1.6 x 10(-3) M. 3. Tryptophan does not interact with the oxidation product of the dopa-oxidase reaction. 4. Neither oxygen nor hydroxyl radicals are involved in the inhibition found in presence of tryptophan. 5. Tryptophan, like dopa, also inhibits tyrosine hydroxylase and dopa oxidase activity of melanosomal tyrosinase and its inhibitory mechanism differs from inhibition due to non-substrate type compounds like cysteine, ascorbic acid. 6. These experiments together with previous findings suggest that the status of tryptophan may be similar to that of dopa in relation to regulation of melanogenesis. PMID- 8224375 TI - Regulation of buccal mucosal calcium channel activity by salivary mucins. AB - 1. The effect salivary mucins on the activity of calcium channel isolated from buccal mucosal cell membranes was investigated. The uptake of 45Ca2+ while only moderately (15%) affected by the intact low and high molecular weight mucin forms, was significantly inhibited, by the acidic low and high molecular weight salivary mucins which evoked 64 and 60% inhibition, respectively. 2. The inhibitory effect of salivary mucins was associated with the sialic acid and sulfate ester groups of the carbohydrate chains, as the removal of either group caused partial loss in the glycoproteins inhibition, and the complete loss in the inhibitory effect occurred following desialylation and desulfation. 3. The channel in the presence of epidermal growth factor (EGF) and ATP responded by an increase in tyrosine phosphorylation of 55 and 170 kDa proteins, and the phosphorylated channels showed a 46% increase in 45Ca2+ uptake. The phosphorylation and the calcium uptake were susceptible to inhibition by a specific tyrosine kinase inhibitor, genistein. 4. The binding of EGF to calcium channel receptor protein was inhibited by the low and high molecular weight acidic mucins, causing 41.2 and 36.1% reduction, respectively. This reduction in binding was dependent upon the presence of sulfate ester and sialic acid groups, as evidenced by the loss of the glycoproteins' inhibitory capacity following removal of these groups. 5. The results for the first time demonstrate that salivary mucins actively participate in the modulation of the EGF-controlled buccal mucosal calcium channel activity expression, a process of importance to the preservation of oral tissue integrity. PMID- 8224376 TI - Properties of enzymes hydrating epoxides in human epidermis and liver. AB - 1. Cytosolic and microsomal epoxide hydrolyzing enzymes of human skin and liver were compared and found to be different. 2. Epidermal and hepatic cytosolic epoxide hydrolases were different in terms of substrate selectivity, pI, inhibitor sensitivity and affinity chromatographic properties. 3. Microsomal epoxide hydrolases had the same pIs but different substrate selectivities. 4. Cytosolic epoxide hydrolase from adults had higher specific activity than that from neonates or cultured epidermis, but lower activity than adult hepatic enzymes. 5. The sizes of cytosolic epoxide hydrolase from epidermis and liver were similar and lower than that from cultured fibroblasts. 6. Cytosolic epoxide hydrolase from all sources shared similar antigenic determinants. PMID- 8224377 TI - Enzyme-to-enzyme tunnelling between phosphoglucoisomerase and phosphofructokinase. AB - 1. Cross-linked and permeabilized rat erythrocytes were incubated for 2-5 min at 37 degrees C in the presence of ATP and either D-[U-14C]glucose 6-phosphate (3 mM) mixed with unlabelled D-fructose 6-phosphate (1 mM) or D-[U-14C]fructose 6 phosphate (1 mM) mixed with unlabelled D-glucose 6-phosphate (3 mM). 2. The contribution of molecules derived from the radioactive ketohexose ester relative to the total amount of newly formed D-fructose 1,6-bisphosphate was lower than the time-related average value for such a relative contribution in the pool of D fructose 6-phosphate. 3. From such a difference, it was calculated that, under the present experimental conditions, 13.1 +/- 2.0% of the molecules of D-fructose 1,6-bisphosphate formed during incubation are directly derived from D-glucose 6 phosphate by a process of enzyme-to-enzyme channelling between phosphoglucoisomerase and phosphofructokinase, rather than originating from the free pool of D-fructose 6-phosphate. 4. A comparable value of 13.2 +/- 3.2% was reached when the process of enzyme-to-enzyme tunnelling was judged from the 3H/14C ratio in D-fructose 1,6-bisphosphate formed by permeabilized erythrocytes exposed for 5-15 min to D-glucose 6-phosphate (3 or 5 mM) mixed with tracer amounts of both D-[1-14C]glucose 6-phosphate and D-[2-3H]glucose 6-phosphate. PMID- 8224378 TI - Effect of membrane phospholipid composition and fluidity on rat liver plasma membrane tyrosine kinase activity. AB - 1. The effect of membrane phospholipid composition and fluidity on tyrosine kinase activity was investigated in rat liver plasma membranes. 2. The phospholipid composition has been modified by in vitro enrichment of plasma membranes with different phospholipids in the presence of lipid transfer proteins and by partial delipidation with exogenous phospholipases A2, C and D and subsequent enrichment with phosphatidylglycerol. 3. Phosphatidylglycerol and dioleoylglycerophosphocholine caused dramatic elevation of this activity, while phosphatidylserine and phosphatidylethanolamine were less effective. Enrichment with dipalmitoylglycerophosphocholine and sphingomyeline reduced tyrosine kinase activity. PMID- 8224379 TI - Enzyme activities and level of SH groups in breast carcinomas. AB - 1. The carcinoma showed higher enzyme activities than the normal mammary tissue. 2. The ratios of glutamate dehydrogenase, glutathione reductase and catalase to lactate dehydrogenase were lower in carcinomas than in normal tissues. Similarly, the ratios of glutamate dehydrogenase, glutathione reductase and catalase to glucose-6-phosphate dehydrogenase were also significantly lower in carcinomas. 3. There were no significant differences in enzyme activities between stages I and II of disease, however in the metastatic tissues, there were significant differences between stages I and II. 4. SH groups were higher in the tissues of cancer patients than in normal tissues. The levels of thiols groups were higher in carcinomas at stage III of disease. PMID- 8224381 TI - Fe parameters and erythrocytic parameters in experimental hyperthyroidism. PMID- 8224380 TI - Species-specific differences in covalently crosslinked complexes of yeast cytochrome c peroxidase with horse and yeast iso-1 ferricytochromes c. AB - 1. The results of chemically crosslinking yeast cytochrome c peroxidase with both horse and yeast iso-1 ferricytochromes c have been studied by a combination of gel electrophoresis and proton NMR spectroscopy. 2. The complexes were formed at a variety of potassium phosphate concentrations ranging from 10 to 300 mM using the water soluble crosslinking agent, EDC (1-ethyl-3-[3-(dimethylamino)propyl] carbodiimide). 3. The primary crosslinking product in both cases is the 1:1 covalent complex, but, for each pair of partner proteins the yield of the 1:1 crosslinked complex varies with the salt concentration. 4. Furthermore, at low salt concentrations the yield of the 1:1 covalent complex involving horse cytochrome c is much larger than the yield of the 1:1 covalent complex formed with yeast iso-1 cytochrome c, whereas at high salt concentrations the situation is reversed. 5. Proton NMR spectroscopy, in combination with gel electrophoresis, provides evidence for the formation of different types of 1:1 complexes for the peroxidase/yeast cytochrome c pair and has been used to study the effect of changes in the solution ionic strength upon both the peroxidases/horse cytochrome c and the peroxidase/yeast cytochrome c complexes. 6. This work indicates that electrostatic interactions between proteins play a dominant role in formation of complexes between cytochrome c peroxidase and horse ferricytochrome c, whereas the hydrophobic effect plays a comparatively larger role in stabilizing complexes between cytochrome c peroxidase and yeast iso-1 ferricytochrome c. PMID- 8224382 TI - Fe parameters and erythrocytic parameters in experimental hypothyroidism. PMID- 8224383 TI - Identification of a membrane interactive protein anchor. PMID- 8224384 TI - Specific immunodetection and purification of G proteins from Sorghum bicolor. PMID- 8224385 TI - Identification of a membrane receptor for the auxin binding protein. PMID- 8224386 TI - Stimulation of guanosine 5'-O-(3-thio)triphosphate binding to higher plant plasma membranes by the mastoparan-analogue, Mas 7. PMID- 8224387 TI - An investigation of tomato extensin. PMID- 8224388 TI - Very low density lipoprotein subfraction apolipoprotein composition. PMID- 8224389 TI - Expression of hormone-sensitive lipase in macrophage foam cells. PMID- 8224390 TI - Domain structure of hormone-sensitive lipase. PMID- 8224392 TI - Mono- and di-acylglycerol concentrations in human plasma in relation to lipoprotein lipase activity. PMID- 8224391 TI - Hydroxypyridinones and desferrioxamine inhibit macrophage-mediated LDL oxidation by iron but not by copper. PMID- 8224393 TI - Purification and partial characterisation of aminopeptidase c from ovine cerebra. PMID- 8224394 TI - Relationship between beta-amyloid precursor protein, pyramidal neurones and astrocytes in human neocortex. PMID- 8224395 TI - Factors affecting the beta-amyloid precursor in PC12 cells. PMID- 8224396 TI - A cortical pyramidal neurone neurotransmitter receptor that may affect beta amyloid precursor protein. PMID- 8224397 TI - Protease "Clipsin" activity and beta-amyloid precursor protein degradation by Alzheimer brain. PMID- 8224399 TI - Metabolism of insect hypertrehalosemic hormone in Blaberus discoidalis cockroaches. PMID- 8224398 TI - Characterisation of putative Drosophila angiotensin converting enzyme cDNA clones. PMID- 8224400 TI - Identification of an ACE-like peptidyl dipeptidase activity in the housefly, Musca domestica. PMID- 8224401 TI - Hydrolysis of cholecystokinin by the human astrocytoma clone D384 and the neuroblastoma line SH-SY5Y. PMID- 8224402 TI - Protein kinase C subtypes associated with the regulation of noradrenaline release in human SH-SY5Y neuroblastoma cells. PMID- 8224403 TI - Inhibitor profile of porcine aminopeptidase W. PMID- 8224404 TI - Characterization of the soluble and membrane-bound forms of porcine angiotensin converting enzyme. PMID- 8224405 TI - Interaction of vacuolar-type H(+)-ATPases with fluorescent organotin-flavone complexes. PMID- 8224406 TI - Role of free radical enzymes in resistance to the dicarboximide fungicides. PMID- 8224407 TI - The reaction of hydrogen peroxide with cytochrome bo from E. coli. PMID- 8224408 TI - Simulation of dioxygen free radical reactions. PMID- 8224409 TI - The analysis of flux in substrate cycles. PMID- 8224410 TI - Cyanide binding to single-electron-reduced fast cytochrome-c oxidase. PMID- 8224411 TI - Flash photolysis of the carbon monoxide compounds of mutant and wild-type cytochrome bo from E. coli. PMID- 8224412 TI - Possible functions of the NADP-linked isocitrate dehydrogenase and H(+) transhydrogenase in heart mitochondria. PMID- 8224413 TI - Conformational studies on heptapeptide analogues of the invertebrate neuropeptide FMRFamide. PMID- 8224414 TI - Synthesis of deltorphin I analogues and preliminary characterisation of mu receptor binding. PMID- 8224415 TI - Molecular interaction of LHC II kinase with petD peptides. PMID- 8224416 TI - Pentapeptide sequences bind to an 80kD Kupffer cell surface protein. PMID- 8224418 TI - Substrate specificity of the insulin receptor tyrosine kinase domain. PMID- 8224417 TI - Interaction of the cytoplasmic domain of phospholamban with calmodulin. PMID- 8224419 TI - Peptide mimetics of nucleotide phosphate binding sites. PMID- 8224420 TI - Conformationally constrained dipeptides as inhibitors of chymotrypsin. PMID- 8224421 TI - Peptides from the N-terminal of link protein are processed differently by chondrocytes and monocytic cells. PMID- 8224422 TI - Chemical modification of N-methyl-D-aspartate receptors from pig brain. PMID- 8224423 TI - Ca2+ binding sites of the ryanodine receptor/Ca2+ release channel of sarcoplasmic reticulum; characterization by fluorescence measurements. PMID- 8224424 TI - Quantitation of type I and III collagen of liver in alloxan-induced diabetic rabbits. PMID- 8224425 TI - Interstitial collagens isolated from lung of rabbits with alloxan-induced diabetes mellitus. PMID- 8224426 TI - Alterations of collagen content in kidney of diabetic rabbits. PMID- 8224427 TI - Characterisation of an endothelin converting enzyme from ECV304 cells, a human endothelial cell line. PMID- 8224428 TI - A differential extraction procedure for studying changes in skeletal muscle proteins. PMID- 8224429 TI - Associations between intermediate filament protein and intermediate filament associated protein, and between intermediate filament associated protein and membrane protein, in wool. PMID- 8224430 TI - Equation of an alpha-helix in a coiled-coil. Distortion of the bond lengths and angles. PMID- 8224431 TI - The intracellular relocation of annexin V in response to the physiological agonist thrombin. PMID- 8224432 TI - Human gene mapping and the polymerase chain reaction. PMID- 8224433 TI - The effect of (+)-catechin, a naturally occurring flavonoid, on benzopyrene oxidation by cells in culture. PMID- 8224434 TI - The regulation of bile acid synthesis in cultured hamster hepatocytes. PMID- 8224435 TI - Erythrocyte sorbitol concentration in relation to blood gas tensions and acid base balance parameters. PMID- 8224436 TI - Sorbitol content and ultrastructure of the aortic tissues of streptozotocin diabetic rats: relation to plasma glucose, glycated hemoglobin and blood sorbitol. PMID- 8224437 TI - Intraepithelial tissues as specific sites for the gamma delta T cells. PMID- 8224438 TI - Does neutral endopeptidase have a role in craniofacial development? PMID- 8224439 TI - Stimulation of phospholipase-D in Epstein Barr virus-transformed human B-cells. PMID- 8224440 TI - Localization of annexins V and VI in the porcine nervous system. PMID- 8224441 TI - Differential responses to histamine and cyclo-pentyladenosine in subclones of the DDT1MF-2 cell line. PMID- 8224442 TI - Effect of unilateral changes in milking frequency on mammary mRNA concentrations in the lactating goat. PMID- 8224443 TI - Metabolism of 5-HT in the nematode Caenorhabditis elegans: identification of a 5 HT N-acetyltransferase that is developmentally regulated. PMID- 8224444 TI - Intracellular cross-talk between receptors coupled to phospholipase C via pertussis toxin sensitive and insensitive G-proteins. PMID- 8224445 TI - The transfer of docosahexaenoic acid from the yolk to the tissues of the chick embryo. PMID- 8224446 TI - The esterification of cholesterol in the yolk sac membrane of the alligator embryo. PMID- 8224447 TI - Neurotoxicity of nicotinamide derivatives: their role in the aetiology of Parkinson's disease. PMID- 8224448 TI - Re-oxygenation induced cardiac damage is prevented by supplementation with vitamin E in vitro but not by modification of dietary vitamin E content. PMID- 8224449 TI - Role of regenerating muscle fibres in aberrant prostaglandin metabolism by dystrophic muscle tissue. PMID- 8224450 TI - Analysis of collagen composition in acne keloids. PMID- 8224451 TI - Elastase levels in small and non-small cell lung carcinoma. PMID- 8224452 TI - Evaluation of "neuron-specific enolase" as a tumor marker for small cell carcinoma of the lung. PMID- 8224453 TI - Tissue and plasma fibronectin level in patients with head and neck carcinoma. PMID- 8224454 TI - Permeability of the respiratory membrane in healthy, non-smoking controls and patients with sarcoidosis and chronic obstructive lung disease. PMID- 8224455 TI - Effects of diet and hormonal status on brown fat lipoprotein lipase activity. PMID- 8224456 TI - Translocation of prepromelittin across the membrane of the endoplasmic reticulum. PMID- 8224457 TI - Cardiac carbohydrate and lipid utilization during late pregnancy. PMID- 8224458 TI - Lactate utilization by the heart in vivo. PMID- 8224459 TI - Antioxidant therapy in children affected by irradiation from the Chernobyl nuclear accident. PMID- 8224460 TI - Expression of annexins in the human neuroblastoma SH-SY5Y. PMID- 8224461 TI - Purification schemes for the constituent domains of cytochrome P450 BM3 in E. coli. PMID- 8224462 TI - Cloning and sequence analysis of a pea seedling amine oxidase gene. PMID- 8224463 TI - Nuclear localisation of corticotrophin releasing hormone (CRH) in transfected CHO K1 cells. PMID- 8224464 TI - Preliminary studies of two active site mutants of galactose oxidase. PMID- 8224465 TI - Membrane trafficking in pituitary cells: studies using a fluorescent dye. PMID- 8224466 TI - Aluminium and Alzheimer's disease: sites of aluminium-binding in human neuroblastoma cells. PMID- 8224467 TI - Aluminium and Alzheimer's disease: electrophoresis of proteins from aluminium treated human neuroblastoma cells. PMID- 8224468 TI - Lysosome preparation from retinal pigment epithelial cells. PMID- 8224469 TI - Differences in the potency of ACAT inhibitors in two assay systems. PMID- 8224470 TI - Raised adenine nucleotide concentrations in erythrocytes of patients with cystic fibrosis. PMID- 8224471 TI - Dissection of the protein phosphorylation cascades involved in insulin and growth factor action. PMID- 8224472 TI - Life style, genetic factors and the risk of heart attack: the apolipoprotein B gene as an example. PMID- 8224473 TI - Structure and function of sphingoglycolipids in transmembrane signalling and cell cell interactions. PMID- 8224474 TI - Molecular recognition in protein families: a database of aligned three dimensional structures of related proteins. PMID- 8224475 TI - Model of the interaction between the UI A small nuclear ribonucleoprotein and UI small nuclear RNA stem/loop II. PMID- 8224476 TI - Molecular recognition in biological systems: from activation to inhibition. PMID- 8224477 TI - Lipocalin structure and function. PMID- 8224478 TI - Molecular mechanism of RNA-phage morphogenesis. PMID- 8224479 TI - Drug therapy for atherosclerosis. PMID- 8224480 TI - The atherosclerotic arterial wall. PMID- 8224481 TI - How to assess regression of the atherosclerotic plaque. PMID- 8224482 TI - Triglyceride-rich particles, high-density lipoprotein and their pharmacological modulation. PMID- 8224483 TI - Anti-oxidants for the treatment and the prevention of atherosclerosis. PMID- 8224484 TI - Oxidized lipoproteins influence gene expression by causing oxidative stress and activating the transcription factor NF-kappa B. PMID- 8224485 TI - Emerging strategies for the treatment of atherosclerosis as seen from the patent literature. PMID- 8224486 TI - Endopeptidase-24.11: putative substrates and possible roles. PMID- 8224487 TI - Targeting of neutral endopeptidase 24.11 in polarized cells. PMID- 8224488 TI - Clinical potential of endopeptidase-24.11 inhibitors in cardiovascular disease. PMID- 8224489 TI - Zinc metallopeptidases: active site structure and design of selective and mixed inhibitors: new approaches in the search for analgesics and anti-hypertensives. PMID- 8224490 TI - The family of pro-hormone and pro-protein convertases. PMID- 8224491 TI - Recent advances on endopeptidase-3.4.24.16. PMID- 8224492 TI - Endothelin-converting enzymes and other families of metallo-endopeptidases. PMID- 8224493 TI - Endocytic regulation of the T lymphocyte co-receptor proteins CD4 and CD8. PMID- 8224494 TI - Endocytosis and intracellular transport of protein toxins. PMID- 8224495 TI - Arguments in favour of endosome maturation. PMID- 8224496 TI - Endosome maturation: insights from somatic cell genetics and cell-free analysis. PMID- 8224497 TI - Cell-free interactions between rat-liver endosomes and lysosomes. PMID- 8224498 TI - Changes in the endocytic compartment in regenerating liver. PMID- 8224499 TI - Ribonucleotide reductases: radical enzymes with suicidal tendencies. AB - Ribonucleotide reductases isolated from E. coli and from L. leichmannii differ considerably in their primary and quaternary structures, as well as in their cofactor requirements. Despite these differences, studies with the wt enzymes and the normal substrate, and with the wt enzymes and a variety of mechanism-based inhibitors, demonstrate amazing mechanistic similarities between the two reductases. Recent studies with five cysteine mutants of both reductases reveal strikingly similar phenotypes, indicating that, despite the differences in the primary structures, the groups involved in catalysis in both enzymes appear to be similar. PMID- 8224500 TI - Crystallographic investigations of ribonucleotide reductase. PMID- 8224501 TI - A model for the catalytic mechanism of prostaglandin endoperoxide synthase. PMID- 8224502 TI - Structure of soybean lipoxygenase-I. PMID- 8224503 TI - Mechanistic pathways in soluble methane mono-oxygenase. PMID- 8224504 TI - Galactose oxidase: molecular analysis and mutagenesis studies. PMID- 8224505 TI - Control of oxidative phosphorylation in liver mitochondria and hepatocytes. PMID- 8224506 TI - Control of oxidative phosphorylation in muscle. PMID- 8224507 TI - Regulation of oxidative phosphorylation in plant mitochondria. PMID- 8224508 TI - Kinetic constraints and oxidative phosphorylation yield in yeast mitochondria. PMID- 8224509 TI - Regulation of the mitochondrial ATP synthase in rat heart. PMID- 8224510 TI - Regulation of electron flux through cytochrome c oxidase: pH, delta pH and fatty acids. PMID- 8224511 TI - Effects of thyroid hormones on oxidative phosphorylation. PMID- 8224512 TI - The role of intramitochondrial Ca2+ in the regulation of oxidative phosphorylation in mammalian tissues. PMID- 8224513 TI - Long-term and short-term changes in mitochondrial parameters by thyroid hormones. AB - In the hyperthyroid state, delta psi m, delta pHm and therefore delta p are increased in rat liver. An enhanced delta p accords with a higher energy output. The subcellular distribution of adenine nucleotides in different thyroid states does not reflect the driving force for mitochondrial adenine-nucleotide translocase (that is delta psi m). Therefore, a change in delta psi m cannot be solely responsible for the postulated stimulation of adenine-nucleotide transport by THs. This is also the case for the changes in delta pHm, and in the subcellular distribution of malate, 2-oxoglutarate and glutamate, that are observed under the influence of THs. T3 induces calcium influx into the liver cell within minutes. It increases respiration and gluconeogenesis with the same kinetics. Therefore, it is suggested that, as with glucagon and vasopressin, calcium is the mediator of these changes. The delta p is increased with T3 and glucagon treatment but not with vasopressin. The changes in delta psi m and delta pHm appear to be the result of the individual actions of these hormones on ATP consuming and ATP-producing reactions. The delta psi p is only increased with T3 treatment. This is related to the different mechanisms of enhancing intracellular calcium that are used by vasopressin, glucagon and T3. PMID- 8224514 TI - Defects of oxidative phosphorylation in man. PMID- 8224515 TI - Differentiation and uteroglobin gene expression by novel rabbit endometrial cell lines. AB - A clear understanding of the mechanism of differentiation of mammalian endometrial epithelia would significantly improve our knowledge of embryo implantation and placentation. An ideal model for such studies would be immortal endometrial epithelial cell lines which express a well characterized, steroid responsive, differentiation-specific gene in vitro. We characterized two cell lines which are temperature-sensitive for differentiation. At the non-permissive temperature (39.5 degrees C), in presence of ovarian steroids, these cells express the gene coding for uteroglobin, a steroid-dependent, immunomodulatory/antiinflammatory protein in the rabbit. In addition, when cultured on artificial basement membrane (Matrigel), in presence of ovarian steroids both cell lines developed organized, tubular structures with lumens, reminiscent of an intact endometrium and secreted 33-fold more uteroglobin than the untreated controls. Thus, these immortal cell lines provide a unique model to study endometrial epithelial cell differentiation and steroid hormone action in vitro. PMID- 8224516 TI - Cloning and sequencing of the sheep pituitary gonadotropin-releasing hormone receptor and changes in expression of its mRNA during the estrous cycle. AB - We have isolated a full length cDNA clone coding for the sheep GnRH receptor (GnRH-R). The amino acid sequence shows greater homology to the human GnRH-R sequence than the two rodent receptors published so far. We have also carried out physiological studies investigating the pattern of expression of the GnRH-R mRNA throughout the estrous cycle. GnRH receptor mRNA and GnRH binding levels were both significantly (P < 0.05) increased over luteal levels up until the time of the preovulatory LH surge, whilst post-surge, a significant (P < 0.05) decline was seen. These changes were related to increased follicle estradiol production in the follicular phase. In contrast, no changes in the abundance of LH beta mRNA were seen throughout the estrous cycle, but the pituitary content of LH significantly (P < 0.05) decreased after the preovulatory LH surge. These results suggest that there is a close relationship between the abundance of GnRH-R mRNA and translation of the GnRH-R in sheep. PMID- 8224517 TI - Angiotensin increases aldosterone synthase mRNA levels in human NCI-H295 cells. AB - Understanding the regulation of aldosterone secretion has been hampered by the lack of a cell culture system that remains chronically responsive to angiotensin stimulation. NCI-H295 cells, cultured from a human adrenocortical tumor, express the three major pathways of adrenal steroidogenesis and produce small amounts of aldosterone during basal culture. We have determined changes in aldosterone production and in aldosterone synthase (AS, P45011B2) mRNA levels in these cells in response to angiotensin II (AII) and forskolin. Culture of NCI-H295 cells with 10(-7) M AII or with 10(-5) M forskolin stimulated aldosterone production and increased AS mRNA levels, though the effect of AII was greater. When cells were cultured with increasing concentrations of AII from 10(-11) through 10(-8) M, a dose-dependent increase in AS mRNA levels paralleled increases in aldosterone production. In view of these findings, these human adrenocortical cells should be useful for exploring mechanisms regulating aldosterone production. PMID- 8224518 TI - The turnover of growth hormone (GH)-binding protein and GH receptor in rabbit and rat. AB - The present study was undertaken to further explore the comparative dynamics of growth hormone-binding protein (GH-BP) in relation to the turnover of the GH receptor (GH-R) in vivo in rabbits and rats. The strategy used was to examine the time course of hepatic GH-R turnover over a 3 h period after cycloheximide treatment, with simultaneous measurements of serum GH-BP level. In the rabbit we sampled multiple liver biopsies and serum samples consecutively from each animal. In the rat, experiments on individual animals were conducted for each time point. In the rat, both liver GH-R and serum GH-BP declined after cycloheximide injection following first-order kinetics. The t 1/2 values for GH-R and GH-BP were 29.7-44.5 and 82.7-119.5 min (95% confidence limits), respectively. A significant positive correlation was found between rat liver GH-R and serum GH-BP (r = 0.85; p < 0.001). In contrast, the decline in rabbit liver GH-R, following cycloheximide treatment was accompanied by simultaneous time-dependent accumulation of serum GH-BP. The t 1/2 for rabbit serum GH-BP accumulation was 30.4-67.6 min. Scatchard analysis of [125I]hGH binding to rabbit GH-BP indicated that the binding capacity increased from 2818 +/- 538 fmol/ml, at time zero, to 5236 +/- 419 fmol/ml following 60 min cycloheximide treatment (p < 0.05). No significant changes in affinity were observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224519 TI - Transcriptional regulation of dog prostate arginine esterase gene by androgens. AB - These studies were designed to define the molecular events involved in the modulation of dog prostate arginine esterase gene expression following short castration intervals and androgen treatment. Arginine esterase enzymatic activity and protein levels decreased about 50% 24 h after castration. Thereafter, a more progressive decrease was observed, resulting in 2-4-fold lower levels in 12-day castrates than in the intact controls. Total prostatic arginine esterase mRNA levels slowly decreased during the first five days after castration but more abruptly thereafter and were about 150-fold lower in 12-day castrated animals. By contrast, in isolated prostatic nuclei, levels of arginine esterase RNA precursors and mature transcripts rapidly fell following orchiectomy, with a 50 70% decrease 24 h after castration. Nuclear run-on experiments confirmed that the latter effects were the result of decreased arginine esterase gene transcription. All these changes could be at least partially reversed by administration of testosterone cypionate. Furthermore, no striking modifications in the proportion of epithelial/stromal cells in the prostatic tissue were observed following orchiectomy. These results show that castration and androgens exert very rapid effects on the gene expression of arginine esterase, and that the regulation occurs at the transcriptional level. PMID- 8224520 TI - Proteins interacting with an androgen-responsive unit in the C3(1) gene intron. AB - The expression of the three genes encoding the components C1, C2 and C3 of prostatic binding protein (PBP) is under androgen control and restricted to the rat ventral prostate. The SstI-PvuII fragment of the first intron of the C3(1) gene displays two binding sites for ubiquitous transcription factors and one for a tissue-specific factor in a 80-bp region upstream of its androgen response element (ARE). The octamer transcription factor 1 (OTF-1) binds to the most distal element (site 1) while a member of the nuclear factor I (NF-I) family recognizes site 2. A third unidentified prostate-specific factor, which also occurs in castrated rats, interacts with the proximal element (site 3). In T-47D cells, both the OTF-1 and the NF-I-like factor can modulate the androgen response of the promoter in a reporter gene construct containing the C3(1) intronic fragment. PMID- 8224521 TI - Inhibition and down-regulation of protein kinase C in cultured atrial myocytes: effects on distribution of specific granules and secretion of atrial natriuretic peptide. AB - Primary cultures of neonatal rat atrial myocytes were subjected to down regulation of protein kinase C (PKC) and to inhibition of PKC activity. The effects on secretion of atrial natriuretic peptide (ANP), and the intracellular distribution of ANP-containing specific granules and their association to microtubules, were investigated. Treating the cultures with inhibitors of PKC, staurosporine or H7, a translocation of ANP-containing specific granules from the perinuclear sarcoplasm to the periphery of the myocytes was observed, and furthermore, secretion of ANP was significantly decreased. The microtubule network were not structurally affected by the PKC inhibitors. Down-regulation of PKC by 12-O-tetradecanoylphorbol 13-acetate (TPA) for 12 h was not followed by any alteration of localization of specific granules, and the amount of secreted ANP was still considerable. Treating the down-regulated cultures with staurosporine, secretion of ANP was still significantly reduced. The present results suggest that the decreased ANP secretion and the translocation of ANP containing specific granules in the atrial myocytes following treatment with staurosporine or H7, is mediated through mechanisms not, or only partly, requiring PKC. PMID- 8224522 TI - Subcellular compartmentalization of MCF-7 estrogen receptor synthesis and degradation. AB - Turnover of the estrogen receptor protein was studied by using enucleation of human breast cancer-derived MCF-7 cells, to examine receptor synthesis and receptor degradation in the separated cytoplasmic compartment (cytoplasts) and nuclear compartment (nucleoplasts). Cytoplasts synthesized estrogen receptors as measured by both hormone-binding and immunoassay, while estrogen receptors (but not progesterone or glucocorticoid receptors) were rapidly degraded in nucleoplasts with a half-life of 3-4 h. Little or no degradation of estrogen receptors in cytoplasts was observed under several conditions. Interestingly, MCF 7 cytoplasts contained approximately 15% of the cell's estrogen receptors, which were not 'translocated' by treatment with 17 beta-estradiol before enucleation. We conclude that the estrogen receptor can be synthesized at least to a hormone binding form in the cytoplasm alone without requiring processing in the nucleus, while the nucleus (or perinuclear cytoplasm) is the primary site of degradation of the estrogen receptor protein. In addition, the presence of a population of estrogen receptors that is cytoplasmic but nontranslocatable may need to be considered in the subcellular localization and actions of steroid receptors. PMID- 8224523 TI - Regulation of interleukin-8 gene expression in human endometrial cells in culture. AB - In this study, we investigated the regulation of interleukin-8 (IL-8) gene expression in separated endometrial stromal and epithelial cells of human endometrium. This research was conducted as part of an analysis of the role of these cells in regulating the recruitment of leukocytes to the endometrium. Well characterized model systems were used to study the regulation of endometrial IL-8 gene expression, namely, stromal cells in monolayer culture after first passage and glandular epithelium in primary culture. The levels of IL-8 mRNA and the accumulation of immunoreactive IL-8 in the medium of endometrial stromal cells is culture increased in a time- and concentration-dependent manner upon treatment with IL-1 alpha, tumor necrosis factor-alpha, or serum. The effects of IL-1 alpha plus serum on IL-8 mRNA levels were at least additive. Serum treatment caused a modest stimulation of IL-8 gene transcription (evaluated after 6 h of treatment) in endometrial stromal cells, but serum also acted in these stromal cells to prolong the half-life of IL-8 mRNA by more than 2.5-fold. The regulation of the levels of IL-8 mRNA in endometrial epithelial cells is distinctly different from that in stroma. First, the levels of IL-8 mRNA in non-treated epithelial cells in serum-free medium were much greater than those in stromal cells under similar conditions. Second, whereas the levels of IL-8 mRNA in endometrial epithelial cells also increased in response to serum and to IL-1 in the absence of serum, in the presence of serum, IL-1 treatment caused no appreciable change in the levels of IL-8 mRNA as was the case in endometrial stromal cells. PMID- 8224524 TI - Targeting of pro-opiomelanocortin to the regulated secretory pathway may involve cooperation between different protein domains. AB - The structure of pro-opiomelanocortin (POMC) can be divided into three main domains: an NH2-terminal domain formed by the NH2-terminal glycopeptide and the joining peptide, a central domain corresponding to the adrenocorticotropin sequences and a COOH-terminal domain containing the beta-lipotropin sequences. Expression of POMC in neuroendocrine cell lines such as the mouse neuroblastoma Neuro2A cells results in its targeting to the regulated secretory pathway of these cells. Intracellular targeting of proteins along non default pathways are widely believed to involve the recognition of specific structural features by a sorting machinery. To understand the nature of the signal involved in targeting prohormone to the regulated secretory pathway, we have constructed mutants of POMC in which sequences from the NH2-terminal, the central and the COOH-terminal domains were deleted and examined the sorting of these mutant POMC molecules in Neuro2A cells by immunofluorescence and immunoelectron microscopy. Our results indicate that POMC NH2-terminal glycopeptide or beta-LPH domain do not contain sufficient information for targeting to the regulated pathway since these peptides are not sorted to secretory vesicles when expressed in Neuro2A cells: Similarly, the ACTH domain does not contain essential targeting information since POMC mutants lacking these sequences were sorted to secretory vesicles. Mutant POMCs containing the sequences of more than one of the main protein domains were, however, correctly targeted to the regulated secretory pathway. Our results indicate that POMC is not targeted to the regulated secretory pathway through recognition of a unique continuous 'molecular address'.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224525 TI - The differential effects of protein kinase C activators and inhibitors on rat anterior pituitary hormone release. AB - We investigated the possibility that various protein kinase C (PKC) activators and inhibitors may differentially affect luteinizing hormone (LH) and growth hormone (GH) release from rat anterior pituitary tissue, incubated in vitro. Activators of PKC induced LH release with the following order of potency: mezerein > phorbol 12,13-dibutyrate (PDBu). Mezerein and PDBu were equipotent on GH release. A range of PKC inhibitors (including compounds highly selective for PKC) potently and completely inhibited PKC activator-induced LH and GH release. Chelerythrine and 1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H7) were less potent inhibitors of PDBu-induced GH release than of LH release. A component of PDBu- and mezerein-induced LH release was inhibited by H7 with high potency, but a second H7-insensitive component was detected. Mezerein- and PDBu induced GH release consisted of an H7-resistant component only. When the regulatory domain of PKCs from different sources was investigated by displacement of [3H]PDBu binding, the affinity for mezerein was 3-5-fold greater than that for PDBu at PKCs from cerebral cortex, lung and alpha and beta isoforms extensively purified from brain. Anterior pituitary PKCs were unusual in showing closely matched affinity for mezerein and PDBu, reminiscent of their equivalent potency on GH release. In order to investigate the potency of the catalytic domain inhibitor H7 on PKCs from different sources, enzyme activity assays were carried out on partially purified cytosolic PKCs from midbrain and anterior pituitary and on extensively purified PKC alpha and PKC beta. The Ca(2+)-independent component of PDBu-induced (phosphatidylserine-dependent) activity from anterior pituitary alone showed unusually low potency of inhibition by H7 but was potently inhibited by staurosporine and Ro 31-8220. In contrast, the Ca(2+)-dependent PKC activity in anterior pituitary was inhibited by H7, staurosporine and Ro-31-8220 with high potency as in all other preparations. These results are consistent with the presence and active role in secretion of pharmacologically distinct forms of PKC (or PKC-like kinases) in rat anterior pituitary cells. PMID- 8224526 TI - Regulation of CYP11A gene expression in bovine ovarian granulosa cells in primary culture by cAMP and phorbol esters is conferred by a common cis-acting element. AB - Production and secretion of steroid hormones throughout the ovarian cycle occurs in a highly episodic and coordinated fashion that requires precise and finely tuned regulatory mechanisms. The regulation of ovarian steroidogenesis by the gonadotropin follicle stimulating hormone (FSH) and luteinizing hormone (LH) as well as by other factors occurs, at least in part, at the level of expression of the genes encoding steroidogenic enzymes. The present study is aimed at the elucidation of regulatory mechanisms by which cyclic adenosine monophosphate (cAMP) and protein kinase C regulate cytochrome P450scc (CYP11A) gene expression in bovine granulosa cells in primary culture. As a first step we characterized the bovine granulosa cell cultures with regard to regulation of P450scc activity and mRNA levels upon treatment with forskolin and/or the phorbol ester TPA. Forskolin, a potent stimulator of cAMP generation, increased both progesterone secretion and P450scc mRNA levels. In contrast, treatment with TPA alone decreased both basal progesterone production and P450scc mRNA accumulation. Co treatment with forskolin and TPA decreased progesterone and P450scc mRNA levels as compared to forskolin treatment alone. The possibility that TPA interfered with the forskolin-stimulated cAMP production could be excluded because simultaneous treatment of granulosa cells with TPA and forskolin potentiated the formation of cAMP. In order to identify regulatory sequences within the 5' flanking region of the bovine CYP11A gene, chimeric DNA constructs comprizing regions of the CYP11A gene fused to a beta-globin-derived reporter gene were transfected into granulosa cells in primary culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224527 TI - Dexamethasone stimulates rRNA gene transcription in rat myoblasts. AB - The glucocorticoid analogue, dexamethasone, stimulated RNA synthesis more than two-fold in rat L6 myoblasts, without affecting the rate of cell proliferation. Treatment of myoblasts for 24 h with 10(-7) M dexamethasone resulted in a 30% increase in the cellular RNA level. More than a two-fold stimulation of pre-rRNA gene transcription by dexamethasone, as measured in isolated nuclei and by cell free transcription, was accompanied by a corresponding increase in pre-rRNA levels. Co-incubation of myoblasts with cycloheximide and dexamethasone did not affect the enhanced pre-rRNA gene transcription demonstrating that de novo protein synthesis was unnecessary to manifest the dexamethasone effect on rDNA transcription. Support for this conclusion is provided by the finding that the levels of UBF1 and UBF2, rDNA upstream binding transcription factors, remain unchanged. The glucocorticoid antagonist RU38486 [11 beta-(4 dimethylaminophenyl)17 beta-hydroxy-17 alpha-(prop-1-ynyl)estra- 4,9-dien-3-one] inhibited the dexamethasone-stimulated rRNA gene transcription suggesting that the glucocorticoid receptor is involved in the response mechanism. PMID- 8224528 TI - Connectional distinction between callosal and subcortically projecting cortical neurons is determined prior to axon extension. AB - In adult rats, layer 5 cortical neurons send axons through the corpus callosum to contralateral cortex or through the internal capsule to subcortical targets, but individual neurons reportedly do not have both connections. Here we confirm this adult separation and address whether it develops by extension of axon collaterals to both sets of targets with later elimination of one or the other (a phenomenon common in developing cortex) or by initially selective axon outgrowth. Retrograde tracers Fast Blue and Diamidino Yellow were injected in the subcortical path at the pyramidal decussation and in the contralateral cortex, respectively, of adult and newborn rats. In 16 adults, no cortical neurons were double-labeled, indicating that none project to both sites. In 17 neonates, hundreds of thousands of layer 5 neurons were single-labeled in each brain, but only one was double labeled. In cases in which the injections to one of the two targets was delayed, again, no double-labeled cells were found. These results indicate that the connectional distinction found in adults is not achieved by the elimination of long transient callosal or subcortical collaterals. To determine if shorter transient collaterals are extended by callosal neurons into the internal capsule, i.e., the subcortical pathway, we injected DiI into one cortical hemisphere of aldehyde-fixed Embryonic Day (E)19 and E21 brains. Two types of axons are seen in the white matter of the cortex opposite the injection: those which turn and extend through the callosum and those which branch, sending collaterals to both ipsilateral and contralateral cortex. Rarely were collaterals seen to extend into or toward the internal capsule. [3H]Thymidine birthdating studies show that layer 5 callosal and subcortically projecting neurons are generated at the same stage of corticogenesis. Thus, from early stages of axon extension, callosal and subcortically projecting cells are distinct classes of neurons and, responding differentially to cues present in cortex, initiate growth toward class-specific and nonoverlapping sets of targets. We conclude that the distinction between the two projection classes in inherent to them and is likely to be determined at an early stage of cortical development, prior to neuronal migration from the neuroepithelium. PMID- 8224529 TI - Fibroblasts promote Schwann cell basal lamina deposition and elongation in the absence of neurons in culture. AB - We and other investigators have repeatedly observed that, although purified populations of Schwann cells cultured in the absence of neurons immunostain for some basal lamina constituents, assembled basal lamina is not seen ultrastructurally. In this study we show that purified rat Schwann cells cultured with either fibroblasts or diffusible substances from fibroblasts acquire basal lamina; this basal lamina is most robust and continuous when Schwann cells and fibroblasts are cultured together. The diffusible fibroblast factor(s) is ascorbate-dependent and heat-sensitive and exhibits a molecular weight greater than 30-50,000. The addition of either a combination of basal lamina components or purified laminin also results in the deposition of lengths of basal lamina. Extracellular matrix elaboration promoted by fibroblast factors is linked to dramatic changes in the morphology of Schwann cells; the cells become elongated and aligned with respect to each other. The addition of purified laminin partly mimics this effect. These changes may represent an early response of the Schwann cell to the acquisition of basal lamina preceding further differentiation in vivo. Our observations resemble those reported for other epithelial cell fibroblast interactions and thus reaffirm the identity of the Schwann cell as an epithelial cell sensitive to the influence of mesenchymally derived factors. We propose that fibroblast stimulation of Schwann cell extracellular matrix deposition probably occurs in normal development with axons and in abnormal situations without axons, such as peripheral nerve injury and tumor formation. PMID- 8224530 TI - Development of the excitation-contraction coupling apparatus in skeletal muscle: association of sarcoplasmic reticulum and transverse tubules with myofibrils. AB - The formation and maintenance of the highly regular organization of membrane systems and proteins in striated muscle require specific membrane-membrane and membrane-cytoskeleton interactions. The development of T-tubules and sarcoplasmic reticulum (SR) was followed in gastrocnemius muscle fibers from chicken embryos between 12 days (E12) and 21 days (E21) of incubation, with particular attention to their relationship with one another and with the myofibrils. The fluorescent lipid analog DiIC16[3] was used to label either the external membranes (plasmalemma and transverse (T)-tubules) or the internal SR in living and fixed muscle. Short membrane invaginations can first be seen in fibers at E14, and at E15 longitudinal T-tubules appear in the periphery of the fibers. A complex network of T-tubules filling the whole fiber diameter develops suddenly at E16. In contrast, SR is abundant at the earliest observed stage (E12) and forms regularly spaced cross striations located at the I-Z-I bands. These correspond to a specific accumulation of smooth membranes around the Z-discs seen in electron micrographs. While SR is specifically associated with the newly formed myofibrils in the periphery of the fibers, the disposition of early T-tubules shows little specific relationship to either SR or the myofibrils. However, electron microscopy shows that junctions between T-tubules and SR are formed during this period (Takekura and Franzini-Armstrong, submitted for publication). Junctions do not acquire a specific relation to the myofibrils until around hatching when triads begin to reorganize into their mature location, the A-I junction. These findings indicate three key events in the organization of T-tubules and SR in the sarcomeres: (1) early SR/Z-line interactions independent of T-tubules; (2) SR/T tubule interactions to form the triad junctions, independent from the myofibrils; and (3) the late association of the junctional complexes with the myofibrils at the A-I border. PMID- 8224531 TI - Deep cytoplasmic rearrangements in ventralized Xenopus embryos. AB - Following fertilization in Xenopus, dramatic rearrangements of the egg cytoplasm relocalize maternally synthesized egg components. During the first cell cycle the vegetal yolk mass rotates relative to the egg surface, toward the sperm entry point (SEP) (J. P. Vincent, G. F. Oster, and J. C. Gerhart, 1986, Dev. Biol. 113, 484-500), while concomitant deep cytoplasmic rearrangements occur in the animal hemisphere (M. V. Danilchik and J. M. Denegre, 1991, Development 111, 845-856). In this paper we examine the role of vegetal yolk mass rotation in producing the animal cytoplasmic rearrangements. We inhibited rotation by uv-irradiating embryos during the first cell cycle, a treatment that yields an extremely ventralized phenotype. Both uv-irradiated embryos and unirradiated control embryos show cytoplasmic rearrangements in the animal hemisphere during the first cell cycle. Cytoplasmic rearrangements on the SEP side of the embryo associated with the path of the sperm pronucleus, plus a swirl on the anti-SEP (dorsal) side, are seen, whether or not yolk mass rotation has occurred. This result suggests a role for the expanding sperm aster in directing animal hemisphere cytoplasmic movements. In unirradiated control embryos the anti-SEP (dorsal) swirl is larger than that in uv-irradiated embryos and often extends into the vegetal hemisphere, consistent with the animal cytoplasm having been pulled dorsally and vegetally by the sliding vegetal yolk mass. Thus the yolk mass rotation may normally enhance the dorsalward cytoplasmic movement, begun by the sperm aster, enough to induce normal axis formation. We extended our observations of unirradiated control and uv-irradiated embryos through early cleavages. The vegetal extent of the anti-SEP (dorsal) swirl pattern seen in control embryos persists through the early cleavage period, such that labeled animal cytoplasm extends deep into dorsal third-tier blastomeres at the 32-cell stage. Significantly, in uv-irradiated embryos, which have not undergone vegetal rotation, most of this labeled material remains more equatorial. PMID- 8224532 TI - The egg nucleus regulates the behavior of sperm nuclei as well as cycling of MPF in physiologically polyspermic newt eggs. AB - The possible role of the egg nucleus in regulating the behavior of sperm nuclei and the cycling of maturation-promoting factor (MPF) was investigated in the physiologically polyspermic eggs of the newt Cynops pyrrhogaster. Many sperm entered all areas of the egg, but only one sperm pronucleus, the principal sperm pronucleus, moved to the center of the animal hemisphere to form a zygote nucleus with the egg pronucleus. All sperm and egg pronuclei synthesized DNA, but the zygote nucleus completed the synthesis of DNA 0.5-1 hr earlier than the accessory sperm nuclei. Entrance into M phase by the accessory sperm nuclei was delayed to a greater and greater extent with increasing distance of these nuclei from the zygote nucleus. When DNA in the egg nucleus was damaged by uv irradiation, not only were both prophase and M phase in the zygote nucleus prolonged, but also the MPF cycle was delayed. Some accessory sperm nuclei in the animal hemisphere escaped degeneration to form additional bipolar spindles, so that delayed multipolar cleavage occurred. The MPF activity in the vegetal hemisphere was less than 25% of that found in the animal hemisphere at M phase in normally fertilized eggs. The levels of immunologically detectable proteins that contained the sequence PSTAIR in vegetal hemispheres were less than 25% of those in animal hemispheres. These results indicate that the egg nucleus of the Cynops egg is involved in the control of the activation of MPF and that the accessory sperm nuclei in the vegetal hemisphere degenerate as a result of the lack of components that are indispensable for entry into M phase. PMID- 8224533 TI - Deep cytoplasmic rearrangements in axis-respecified Xenopus embryos. AB - In fertilized eggs of the frog Xenopus, the vegetal yolk mass rotates away from the future dorsal side (J. P. Vincent and J. Gerhart, 1987, Dev. Biol. 123, 526 539), and a major rearrangement of the deep animal hemisphere cytoplasm produces a characteristic swirl in the prospective dorsal side (M. V. Danilchik and J. M. Denegre, 1991, Development 111, 845-856). The relationship between this swirl and determination of the dorsal-ventral axis was further investigated by attempting to experimentally separate the positions of the swirl and the dorsal-ventral axis. Eggs were obliquely oriented in the gravity field to respecify the direction of yolk mass rotation and the position of the dorsal-ventral axis. When yolk mass rotation occurred in the absence of a sperm, as in activated eggs, a swirl pattern formed on the side away from which the yolk mass had rotated. In fertilized eggs tipped with the sperm entry point (SEP) down or to the side, swirl patterns were always found to form on the side away from which the yolk mass was displaced. However, in eggs tipped SEP up, in which the yolk mass was forced to rotate away from the SEP, more complicated rearrangements were observed in addition to the rotation-oriented swirl. Because the direction of yolk mass rotation was found to be influenced by both gravity and the actual position of the SEP in obliquely oriented eggs (SEP to the side), such complicated rearrangement patterns may result from opposing forces generated by both yolk mass rotation and the expanding sperm aster. Thus, except in cases in which the influences of SEP position and unit gravity opposed each other, it was not possible to experimentally separate the position of the deep cytoplasmic swirl from the direction of yolk mass rotation, and therefore the position of the prospective dorsal side. PMID- 8224534 TI - In the egg of the ascidian Phallusia mammillata, removal of external Ca2+ modifies the fertilization potential, induces polyspermy, and blocks the resumption of meiosis. AB - The fertilization potential of the Phallusia mammillata egg has been found to consist, in natural or standard artificial sea water (NSW and ASW), of an initial sperm-triggered rapid depolarization comprising two distinct successive components, followed by a phase of membrane depolarization comprising a plateau and two series of membrane potential oscillations. These oscillations occur during meiotic divisions and they end just before formation of the second polar body. In the present work, the effects of removing external Ca2+ on the fertilization potential time course on the rate of polyspermy and on the resumption of meiosis of the P. mammillata egg were explored. Eggs inseminated in Ca(2+)-free ASW usually responded by several successive sperm-induced electrical responses strikingly different from that obtained in standard ASW: removing external Ca2+ during insemination and the initial depolarizing shift (free (Ca2+), 5-11 microM in Ca(2+)-free ASW) and during the subsequent period of the electrical responses (free (Ca2+), 0.2 microM in Ca(2+)-free ASW), completely suppressed both the second component of the initial depolarizing shift, the plateau, and membrane oscillations of the depolarization phase, did not impair sperm entry, but always blocked the resumption of meiosis. Under these conditions eggs were polyspermic (91%, n = 22, c.l. 69-99%). Lastly, only a short period of contact (approximately 20 sec at minimum) with a physiological concentration of external Ca2+ (12 mM) at the onset of the fertilization potential was necessary for the egg to resume meiosis and then to undergo complete embryonic development. These results indicate that in the P. mammillata egg, the second component of the initial depolarizing shift involves voltage-gated Ca2+ channels. The possibility is discussed that external Ca2+ ions might contribute to the activation of the P. mammillata egg by providing a source for an increase of (Ca2+)i and/or for the reloading of the Ca2+ internal stores. PMID- 8224535 TI - Analysis of cell cycle progression during the development of Dictyostelium and its relationship to differentiation. AB - Labeling with the thymidine analog 5'-bromodeoxyuridine has been used to follow cell cycling during development of the cellular slime mould Dictyostelium discoideum. The results show that cycling continues during development and that 75% of all cells from axenically grown cultures go through S-phase. Although cell cycle progression is temporarily blocked in all cells during early aggregation, it is specifically reinitiated in prespore cells later in development. Prestalk cells do not reenter the cycle. Pulse chase experiments indicate a close correlation between cell cycle phase at the onset of development and differentiation fate. Cells in S-phase and early G2 differentiate preferentially into prestalk cells; cells in the later part of G2 differentiate into prespore cells. PMID- 8224536 TI - Neuronal expression of regulatory helix-loop-helix factor Id2 gene in mouse. AB - Id-like helix-loop-helix (HLH) proteins, which lack a basic DNA binding domain, have been suggested to serve as general inhibitors of differentiation. We present data that Id2 is expressed in specific cell types during neurogenesis and in the adult. At early stages of neurogenesis, Id2 is expressed in the ventricular zone of neuroepithelium. After the first neuronal populations are born, the expression of Id2 is down regulated in neuroepithelial cells and continues to be high in Purkinje cells of the cerebellum, in mitral cells of the olfactory bulb, and in layers 2, 3, and 5 of the cerebral cortex. In neuronally differentiating cell lines, the Id2 expression is up regulated (PCC7), down regulated (NG108), or unchanged (N18) during differentiation. Analyses of promoter sequences of the Id2 gene identify the region which is responsible for the down regulation of transcription during neuronal differentiation. Our data indicate that Id2 has different functions in different cell types during neurogenesis. PMID- 8224537 TI - Regional specification during embryogenesis in the articulate brachiopod Terebratalia. AB - The timing of regional specification during embryogenesis in Terebratalia transversa has been examined by isolating animal, vegetal, and lateral halves at developmental stages ranging from prior to fertilization through late gastrula. Animal halves isolated from prior to fertilization through late blastula stages did not gastrulate and did not form endoderm. When these animal halves were isolated from prior to fertilization through early blastula, they formed only ciliated vesicles; however, similar halves from late blastulae frequently formed the ectodermal components of the three larval lobes. Vegetal halves isolated prior to fertilization through late blastula gastrulated, formed endoderm, and differentiated most larval structures; however, comparable isolates from late gastrulae showed a marked decline in the ability to form anterior larval structures. Animal halves isolated at this stage showed a corresponding decline in the ability to form posterior larval structures. These results indicate that during oogenesis, regional differences arise so that the animal half of the oocyte has a much greater ability to form anterior ectodermal structures than do other regions of the oocyte, while only the vegetal region has the capacity to gastrulate. Inductive signals produced by the vegetal region of the embryo act on the animal half to give it the capacity to form ectodermal components of the three larval lobes. This vegetal-inductive influence has played a major role in regional specification by the late blastula stage of development. Pairs of lateral halves isolated from the same oocyte or embryo from prior to fertilization through late blastula frequently gastrulate, form endoderm, and proceed to form all or most of the larval lobes. The late blastula is the first developmental stage where there is a marked decline in the ability of pairs of lateral halves to differentiate larval structures and is the first stage where pairs of halves resembling dorsal and ventral halves appear. This suggests that the axis of bilateral symmetry is also specified by the late blastula stage of development. PMID- 8224538 TI - Chromatin transitions during early Xenopus embryogenesis: changes in histone H4 acetylation and in linker histone type. AB - We describe major transitions in the type and modification of chromatin associated proteins during the early development of Xenopus laevis. Histone H4 is stored in the diacetylated form in the egg and is progressively deacetylated during normal development. If histone deacetylases are inhibited with sodium butyrate, hyperacetylated histone H4 only accumulates after the mid-blastula transition. The type of linker histone in chromatin also changes during embryogenesis, from predominantly the B4 protein at the mid-blastula transition to predominantly histone H1 at the end of gastrulation. These transitions in chromatin composition correlate with major changes in the replicative and transcriptional activity of embryonic nuclei. PMID- 8224539 TI - Functional heterologous gap junctions in Fundulus ovarian follicles maintain meiotic arrest and permit hydration during oocyte maturation. AB - The physiological significance of heterologous gap junctions between granulosa cells and the oocyte was investigated in late vitellogenic ovarian follicles of the teleost Fundulus heteroclitus. Lucifer Yellow injected into the oocyte readily passed to the overlying granulosa cells, demonstrating effective dye coupling. Passage of the fluorescent dye, and hence intercellular communication, was inhibited both by the tumor-promoting phorbol ester phorbol 12-myristate 13 acetate (PMA) and by 1-octanol, known uncouplers of gap junctions in a variety of invertebrate and vertebrate cell types. Octanol alone also initiated resumption of meiosis in follicle-enclosed oocytes, indicating that granulosa cells normally maintain meiotic arrest, as apparently occurs in mammalian and amphibian follicles. Both PMA and octanol also consistently inhibited the hydration process that normally accompanies meiotic maturation. These results support a previously suggested hypothesis that K+, which is the primary osmotic effector for oocyte hydration, is translocated via gap junction from granulosa cells to the maturing oocyte. PMID- 8224540 TI - Cell-specific expression in the silkmoth follicle: developmental characterization of a major chorion protein, its mRNA and gene. AB - Choriogenesis (eggshell formation) within the silkmoth Antheraea polyphemus proceeds in parallel for the two major subpopulations of follicle cells, diverging only during the very late period when aeropyle crown surface structures form in one region but not in the other. Correlated with their appearance is the synthesis of a set of region-specific proteins. In this report, aeropyle crowns are physically isolated and their protein composition is shown to consist of those same region-specific proteins. A cDNA clone, called pcvl 16, has been selected and shown to encode a lamellar-forming, aeropyle crown-specific protein, probably of the previously described C3,4 group. These conclusions are based on hybrid-selected translation, Northern analysis, and sequence analysis. pcvl 16 was used to isolate two distinct cloned copies of the 16 gene. Both 16 genes are closely paired with another region-specific gene but the proximity of the two gene pairs to each other is uncertain. Non-region-specific chorion genes expressed at earlier times in choriogenesis surround the 16 gene pairs, suggesting that cis sequences necessary for regionalized expression may be closely linked to coding sequences. To test this hypothesis, 5'-flanking sequences from eight region-specific genes are compared and shown to share two oligonucleotide sequences. One is a known regulatory element found in virtually all moth and fly chorion genes examined. The other, located just upstream from the TATA box, is not found in non-regionally expressed chorion genes and, thus, is a candidate for specifying regional expression. PMID- 8224541 TI - MRC-5, human embryonic lung fibroblasts, induce the duplication of the developing chick limb bud. AB - The anteroposterior (A-P) axis pattern of the chick limb is likely to be controlled by a small region of mesenchyme cells at the posterior margin of the limb bud (ZPA, zone of polarizing activity). In this study, we found that MRC-5 fibroblast cells had the capacity for duplicated-pattern formation of the chick limb along the A-P axis when grafted to the anterior region of the limb bud. MRC 5 cells were effective only during pre-limb bud stages, and the leg bud was more responsive than the wing bud. Grafted cells remained at the base of the limb bud when limb development proceeded. These results suggest that the products of MRC-5 cells are involved in three possible processes of duplicated-pattern formation: induction of the polarizing activity; maintenance of this activity, which is present weakly at pre-limb bud stages; and determination of A-P axis as the ZPA factor(s). By allowing the use of a small number of cells of embryonic tissues with polarizing activity, the analysis of duplicate formation with the MRC-5 cell line provides a powerful tool for elucidating the molecular nature of the ZPA. PMID- 8224542 TI - Structural characterization and expression analysis of the Neurospora conidiation gene con-6. AB - The gene con-6 of Neurospora crassa is expressed during the formation of asexual spores (conidia), but it is not expressed in mycelium. con-6 mRNA appears upon induction of conidiation and reaches high levels at the late stages of conidiation, and in mature conidia. The CON6 polypeptide and a CON6-beta-Gal fusion protein were present at high levels only in free conidia. Shortly after spore germination con-6 mRNA disappears and the CON6 polypeptide is degraded. CON6 is a small, hydrophilic polypeptide containing a repeat sequence; it not homologous to any known protein but has features resembling the late embryogenesis abundant proteins of maize. Inactivation of con-6 by the repeat induced point mutation process had no demonstrable effect on formation or germination of conidia. Upstream sequence comparisons for con-6 and other con genes identified a common potential regulatory sequence, designated CRS-B. DNA mobility shift analyses with cell extracts identified a factor that bound to synthetic DNA fragments containing this sequence. This binding factor was present in mycelium but not in conidiating cultures. Experiments with independent integrated con-6'-'lacZ translational fusions revealed substantial variability of expression among transformants carrying identical fusion constructs: This variability may be due to the differential methylation of transformant DNA noted by others. PMID- 8224543 TI - Competence prepattern in the animal hemisphere of the 8-cell-stage Xenopus embryo. AB - Activins are known to be potentially important regulators in Xenopus developmental processes. It has been shown that activins exist maternally in the egg and can induce mesodermal tissues in blastula animal cap explants. However, the blastula ectoderm is known to possess a predisposed local response pattern to activin, and the process of the prepatterning is not understood. We isolated animal hemispheres from late 8-cell-stage embryos and treated them briefly with activin A. Expression of the muscle-specific actin gene was induced after a 30 min activin treatment, even when it was followed by treatment with follistatin, an activin-specific binding protein. This suggests that the animal-half blastomeres become competent to activin A before the 16-cell stage. In the normal embryo, the 8-cell-stage animal dorsal blastomeres populate neural ectoderm and most of the dorsal lip of the gastrula blastopore, the region of Spemann's organizer, and are the major progenitor for dorsal mesodermal tissues. When the dorsal and ventral animal-half blastomeres of the 8-cell stage were isolated and treated with activin independently, significant differences in tissue differentiation were observed. Dorsal blastomeres gave rise to trunk and tail structures containing dorsal mesoderm, whereas the ventral blastomere explants formed spheres containing solely ventral mesoderm. Further, both muscle actin transcription and goosecoid transcription were induced primarily in dorsal blastomeres. Our results suggest that a competence prepattern of response to activin exists as early as the 8-cell stage. PMID- 8224544 TI - Choriogenesis in the Lepidoptera: morphogenesis, protein synthesis, specific mRNA accumulation, and primary structure of a chorion cDNA from the gypsy moth. AB - Choriogenesis in silkmoths has been used as a model for relating developmental processes and evolutionary change. We now describe parallel studies in a nonsilkmoth lepidopteran species, the gypsy moth. Choriogenesis is described at the levels of ultrastructure, protein composition and synthesis, and specific mRNA accumulation. One complete chorion cDNA sequence is presented and features of its primary structure are discussed. This sequence is shown to be homologous to those of silkmoths. PMID- 8224545 TI - A clonal analysis of secondary mesenchyme cell fates in the sea urchin embryo. AB - The secondary mesenchyme cells (SMCs) give rise to most of the mesoderm of the sea urchin embryo. Although the early embryonic lineage of these cells has been described, the mechanisms that cause SMCs to become restricted to a particular mesodermal cell fate are unknown. To begin to address this question, we performed a clonal analysis of the fates of SMC precursors in the vegetal plate by labeling single cells with the fluorescent dye DiI (C18). Our data show that some presumptive SMCs remain pluripotent at the late blastula stage, since some cells labeled at this stage gave rise to more than one mesodermal cell type. Surprisingly, however, most labeled cells gave rise to homogeneous clones composed of a single cell type. This observation indicates that either many SMC precursors are restricted in their fate before the start of gastrulation or that all the progeny of a single vegetal plate cell are influenced by the same instructional signals during gastrulation, despite the cell divisions and extensive cell movements that occur during this time. The percentage of clones composed of a single cell type increased during the blastula stage, supporting the view that the process of SMC fate specification begins before the onset of gastrulation. PMID- 8224546 TI - A decreased number of primordial germ cells and the small numbers and reduced sizes of germinal granules in the periodic albino mutant of Xenopus laevis. AB - Light and electron microscopy were used to examine the cause of the small number of primordial germ cells (PGCs) that were unexpectedly found in periodic albino (a(p)/a(p)) tadpoles of Xenopus laevis. The observations revealed that the volumes of germ plasm were not significantly different between a(p)/a(p) and wild type eggs and that the germ plasm of ap/ap eggs contained a smaller number of germinal granules than are present in wild-type eggs and the granules were also smaller than in the wild-type eggs. It was concluded that the number of PGCs might be primarily determined by the number and size of germinal granules. PMID- 8224547 TI - Survival of extraocular muscle in long-term organotypic culture: differential influence of appropriate and inappropriate motoneurons. AB - The myotrophic effects of a precise matching of motoneuron to target muscle may be particularly apparent during the development of some unique skeletal muscle types, such as the extraocular muscles. To understand how motoneuron-specific factors modulate muscle ontogeny, neonatal extraocular muscle explants were cocultured with either the appropriate midbrain motoneurons or the inappropriate spinal cord motoneurons. Thigh muscle cocultured with spinal motoneurons, which differentiates and survives for several months, served as a control. In short term cultures (during the first 3 weeks in vitro), neonatal extraocular muscle explants developed myotubes that were immunoreactive for myosin, became innervated, and matured in parallel with hindlimb muscle explants. The origin of the motoneurons (midbrain or spinal cord) did not affect extraocular muscle development during the first 3 weeks in vitro. However, in long-term cultures (after the third week in vitro), extraocular muscle that was innervated by the inappropriate spinal motoneurons failed to survive and degenerated. The failure of extraocular muscle to survive when cocultured with the inappropriate motoneurons did not result from an artifact of in vitro conditions. When extraocular muscle primordia were innervated by midbrain explants, many of which contain oculomotor motoneurons, the cultures thrived for > 60 days. We conclude that the trophic requirements of extraocular muscle in organotypic culture are different from those of skeletal muscle isolated from the hindlimb. The novel oculomotor motoneuron-specific interactions that are essential for the survival of extraocular muscle primordia in vitro may prove to be important for the determination of the unusual properties of extraocular muscle in vivo. PMID- 8224548 TI - Star is required for neuronal differentiation in the Drosophila retina and displays dosage-sensitive interactions with Ras1. AB - The photoreceptor cells R8, R2, and R5 are the first cells to initiate neuronal differentiation in the Drosophila eye imaginal disc. We have shown previously that these three cells require Star gene function for proper ommatidial assembly. Due to the embryonic lethality associated with mutations in Star, we have analyzed the consequence of loss of Star function in mosaic eye imaginal discs. Presumptive R8, R2, and R5 cells that lack Star function fail to differentiate neuronally and die a few hours later. Enhancer trap insertions reveal that Star expression in the eye disc is restricted to the developing R8, R2, and R5 cells. Taken together, these data suggest that Star is required for the reception of a signal and/or the execution of a developmental program that leads to the neuronal differentiation of R8, R2, and R5. Star is also required for the formation of wing veins and is expressed in developing veins, suggesting that at least partially overlapping pathways may operate during photoreceptor cell differentiation and wing vein formation. The role of Star in cell-cell signaling is supported by the observation of genetic interactions between Star and mutations that reduce signaling through both sevenless and the Drosophila EGF receptor homologue, including Ras1 and Son of sevenless. PMID- 8224549 TI - Genetic analysis of developmental mechanisms in hydra. XXI. Enhancement of regeneration in a regeneration-deficient mutant strain by the elimination of the interstitial cell lineage. AB - The interstitial cell lineage, including interstitial stem cells, nerve cells, and nematocytes, was eliminated from a regeneration-deficient mutant strain (reg 16) of Hydra magnipapillata. The resultant interstitial cell lineage-free (or "epithelial") reg-16 animals showed a marked enhancement in the ability to regenerate head structures. The epithelial reg-16 polyps regenerated nearly the same number of tentacles as was originally present within 8 days after head removal, while interstitial cell lineage-containing (or "complete") reg-16 polyps restored less than one-third of their original tentacle number under the same conditions. Lateral tissue transplantation was used to examine the head activation and inhibition potentials. The gradients of the two potentials along the body axis of intact epithelial 105 (a wild-type strain) and intact epithelial reg-16 polyps were nearly identical to the gradients in their complete counterparts. The changes of the two potentials occurring after head removal in the epithelial 105 animals were also similar to those in the complete 105 animals. However, the postdecapitation changes in the epithelial reg-16 polyps were different from those in complete reg-16 polyps. The changes in the epithelial reg-16 animals were similar to those observed in wild-type hydra while those of complete reg-16 polyps were highly abnormal. These observations suggest that the phenotypic expression of the genetic defect present in the reg-16 mutant strain is attenuated when the interstitial cell lineage is eliminated from its tissue. The role of the interstitial cell lineage in head regeneration and the nature of the defect present in the reg-16 strain are discussed based on the observations made in this and previous related studies. PMID- 8224550 TI - Developmental progression of DNA puffs in Sciara coprophila: amplification and transcription. AB - DNA amplification for two major DNA puffs (II/2B and II/9A) of the fungus fly Sciara coprophila increases steeply from 17 to 19 days after hatching (18 degrees C), resulting in almost 20-fold more DNA at these loci in mature larval salivary glands than in adult tissues. At 19 days after hatching when gene amplification reaches a plateau, there is a burst in the amount of mRNA encoded by these two DNA puffs. Expansion of the two puffs coincides with the increase in transcription rather than reinitiation of DNA replication. In contrast, an RNA puff (III/9B) undergoes no DNA amplification, and the burst in RNA produced by this locus occurs slightly later, at 20 days after hatching. The progression of cytological puffing and associated molecular events of DNA amplification and transcription correlate with features of the external phenotype of the larvae, especially the number of pigment granules in the eyespots that are the anlage to the adult eyes. The sequential and coordinated regulation of DNA puff replication and transcription is discussed. PMID- 8224551 TI - Cell type regulation in response to expression of ricin A in Dictyostelium. AB - Expression of ricin A in either prespore or prestalk cells of Dictyostelium discoideum results in cell-autonomous lethality. Strains expressing the toxic gene under the control of a prestalk-specific regulatory region fail to culminate or form stalks, but form spores normally. Strains expressing ricin A under the control of a prespore-specific regulatory region form neither spores nor stalks. Regulation of the cell types results in conversion of prestalk cells to prespore cells when the prespore cells are poisoned. The newly converted cells then express ricin A and die. In contrast, we could not detect any significant conversion of prespore cells to prestalk cells when the prestalk cells are poisoned under our experimental conditions. This regulation of cell types suggests that the tendency of prestalk cells to regulate and become prespore cells is inhibited by the already established prespore cells. It appears that prespore cells control prestalk cell regulation by producing an inhibitor of prespore differentiation to which they themselves are insensitive. PMID- 8224552 TI - Behavior of the components of maturation-promoting factor, cdc2 kinase and cyclin B, during oocyte maturation of goldfish. AB - We examined the changes that occurred in the two components of maturation promoting factor (MPF), cdc2 kinase and cyclin B, during oocyte maturation in goldfish, using monoclonal antibodies against the C-terminal sequence of goldfish cdc2 kinase and Escherichia coli-produced full-length goldfish cyclin B. Immature oocytes contained a 35-kDa inactive cdc2 kinase. In addition to the 35-kDa form, a 34-kDa active cdc2 kinase was detected in oocytes undergoing germinal vesicle breakdown (GVBD). Cyclin B was absent in immature oocytes and appeared just before GVBD, coinciding exactly with the appearance of the 34-kDa active cdc2 kinase. Precipitation with p13suc1 beads and anticyclin B antibody revealed that cyclin B formed a complex with cdc2 kinase as soon as it appeared. MPF activation was induced by 1 ng cyclin B after introduction into immature oocytes or oocyte extracts. This corresponds to the amount of cyclin B found in mature oocytes (the concentration in the oocyte is 2 micrograms/ml). These results suggest that MPF activation in fish oocytes is induced by complex formation with preexisting cdc2 kinase and newly synthesized cyclin B during oocyte maturation, a situation differing from that in Xenopus and starfish, in which the cdc2 kinase-cyclin B complex is already present in immature oocytes. Unlike that in Xenopus, an inhibition of protein synthesis in unfertilized mature goldfish oocytes caused a decrease in the cdc2 kinase activity/cyclin B protein level and led to a progression from meiotic metaphase to meiotic anaphase. This result indicates that the mechanisms of maintaining MPF activity in mature goldfish oocytes differ from those in Xenopus. PMID- 8224553 TI - Neuropsychological and neuroradiological study of a case of early-onset Huntington's chorea. AB - The authors report a case of Huntington's disease in an 11-year-old boy with onset at six years of age. The neurological signs and symptoms were midway between the hyperkinetic and rigid forms of chorea. Intellectual development was characterized by a medium-grade deficit. MRI revealed marked atrophy of the head of the caudate nucleus, with diffuse hyperintensity of the putamen. The most characteristic neuropsychological feature was ideomotor apraxia. Neuropsychological and neuroradiological data are discussed in relation to the role of the basal nuclei and frontal cortex in the organization of movement. PMID- 8224554 TI - Delayed parkinsonism associated with hypotension in a child undergoing open-heart surgery. AB - An eight-year-old boy developed acute parkinsonism four days after open-heart surgery for repair of a ventriculo-septal defect. During the procedure he experienced a hypotensive episode which required administration of positive inotropic agents. Complementing the clinical signs of parkinsonism, CT scan showed symmetrical hypodensities in the basal ganglia, and decreased regional cerebral blood flow was demonstrated using 99mTc HMPAO SPECT. These findings were suggestive of a hypoxic-ischaemic insult to the basal ganglia. The child was treated with levodopa/carbidopa and subsequently completely recovered within a follow-up period of eight months. CT scan appearances and cerebral blood flow findings returned to normal. Parkinsonism secondary to a hypoxic-ischaemic insult to basal ganglia in children is a rare but reversible disorder, in contrast to its progressive course which results in severe disability in adults. PMID- 8224555 TI - Startle disease or hyperekplexia. PMID- 8224556 TI - The social needs of adolescents with cerebral palsy. PMID- 8224557 TI - Birth asphyxia: a statement. World Federation of Neurology Group. PMID- 8224558 TI - Alexander's disease. PMID- 8224559 TI - 'New' 3C syndrome. PMID- 8224560 TI - Hip dysplasia in cerebral palsy. PMID- 8224561 TI - In praise of difference. PMID- 8224562 TI - Patterns of neuropsychological function in a low-birthweight population. AB - An investigative analysis was carried out of the neuromotor and cognitive findings in a population-based study of low-birthweight infants (< 1750g) at 4 1/2 years of age to try to improve the understanding of the neuropathological basis of their deficits. Cluster analysis identified groups of children whose performance in the cognitive subscales of the British Ability Scales followed similar patterns, and also differentiated between children in neuromotor competence. Cluster membership correlated highly with language attainment, ability to copy shapes, behaviour as reported by parents and parents' and examiners' perception of attention span. Children in poorer-performing clusters were more likely to be born to mothers who had had a previous perinatal death, and were also more likely to have experienced septicaemia during the neonatal period. These findings are discussed in the light of evidence from the neuropathological and physiological literature. PMID- 8224563 TI - Perinatal hypoxic-ischaemic brain injury: prediction of outcome. AB - Twenty-six term babies with hypoxic-ischaemic brain injury were studied during the neonatal period to evaluate the prediction of outcome to at least one year of age by means of ultrasonography, CT scanning and Doppler ultrasound assessment of cerebral palsy blood flow velocity (CBFV). Adverse outcome was defined as the occurrence of cerebral palsy, developmental delay or death. At follow-up, 17 infants had an adverse outcome (seven died, 10 had disability); the remainder had no detectable impairment. Abnormalities on cranial ultrasound were not, but generalised decreased tissue density on CT scan was, associated with adverse outcome. Abnormal mean CBFV in the middle cerebral artery had no association with outcome, but abnormal mean CBFV in the anterior cerebral artery and a low resistance index in both arteries were significantly associated with adverse outcome. Such information may be used for appropriate counselling of parents of asphyxiated infants. PMID- 8224564 TI - Women with cerebral palsy: obstetric experience and neonatal outcome. AB - The aim of the study was to provide a preliminary basis for advising women with cerebral palsy (CP) who choose to initiate pregnancy regarding the course of parturition and the outcomes for their newborn infants. The authors studied 22 women with CP who had 38 pregnancies at a mean age of 26 years. Eight pregnancies were electively terminated and two resulted in miscarriage. Of the 28 viable pregnancies, one resulted in a preterm stillborn infant and two in preterm liveborn infants. Delivery was vaginal in 18 cases and by cesarean section in nine (one pregnancy was lost to follow-up). Pregnancy outcomes were reassuringly normal in this small, select sample of women with relatively mild CP. Possible effects of pregnancy and childbirth on general adaptive skills or specific child care skills of women with CP could not be assessed in this retrospective study. PMID- 8224565 TI - Parent compliance with therapeutic interventions for children with cerebral palsy. AB - This paper reports the correspondence between different measures of parental compliance and the relationship of these compliance measures to change in hand function after therapy intervention. 72 children with cerebral palsy, aged 18 months to eight years, participated in a clinical trial of neurodevelopmental therapy (NDT) and upper-limb casting; home programmes were part of each intervention. Measures of parental compliance comprised: self-rating of understanding and skill using a questionnaire, therapist's rating of parental compliance, attendances, mean time of cast wear and number of days completed home programme log-book. Parent ratings were the only significant predictor of change in hand function. Parent self-rating was the most useful measure, since it correlated moderately with two other compliance measures and was predictive of change in hand function. PMID- 8224566 TI - Teaching self-feeding skills to patients with Rett syndrome. AB - Few studies have examined the extent to which functional hand-use may be taught to girls with Rett syndrome after those skills have been lost. In this investigation, five females with classic Rett syndrome were taught to feed themselves using a method of prompting and reinforcement. All patients demonstrated improvements in their self-feeding skills. The results are discussed in terms of the factors that may influence the acquisition of skills in girls with Rett syndrome, and directions for future research and proposed. PMID- 8224567 TI - Nutritional status and growth of children with diplegic or hemiplegic cerebral palsy. AB - The growth failure and nutritional status were assessed of 154 children with diplegic or hemiplegic cerebral palsy, aged between two and 17.4 years. Linear growth was significantly reduced compared with norms for healthy children. About 30 per cent of the sample were undernourished, indicated by bodyweight or depleted subcutaneous fat stores at the triceps skinfold site. 8 per cent were overly fat by triceps skinfold and 14 per cent overweight by bodyweight. 23 per cent of the children had stunted growth. Children in the youngest age-group were most at risk for poor nutritional status and delayed growth. These findings suggest that children with diplegic or hemiplegic cerebral palsy are at risk for a variety of abnormalities of growth and nutritional status, and that a growth and nutritional assessment should be conducted periodically so that under- or overnutrition can be corrected when identified. PMID- 8224568 TI - Effect of exogenous pancreatic enzymes on gastrointestinal and pancreatic hormone release and gastrointestinal motility. AB - In clinical practice, exogenous pancreatic enzymes are administered for the treatment of pancreatogenic steatorrhea or with the intention to relieve pain due to chronic pancreatitis. Moreover, a large number of patients take pancreatin (i.e., exogenous pancreatic enzymes) for functional dyspepsia. The effect of exogenous pancreatic enzymes on the enteropancreatic axis is a complex issue. Intraduodenal but not intrajejunal protease activity appears to exert a dose dependent negative feedback on exocrine pancreatic secretion. Only enzymes with a proteolytic activity but not amylase and lipase exert a control on pancreatic secretion. The mechanism responsible for this feedback regulation is debated, but the cholinergic system seems to play a major role. Intraduodenal pancreatic enzymes (pancreatin) lead to an increased release of pancreatic polypeptide but do not affect the release of insulin and glucagon. In addition, pancreatic enzymes have an influence on the release of some gastrointestinal hormones (i.e., cholecystokinin, motilin, gastric inhibitory polypeptide). Neither exogenous nor endogenous pancreatic enzymes seem to play a major role in the regulation of interdigestive gastrointestinal motility. However, an adequate rate of postprandial pancreatic output is required to control gastric emptying. Current knowledge on the effect of exogenous pancreatic enzymes on the enteropancreatic axis, gut peptide release and gastrointestinal motility are updated in the present article. PMID- 8224569 TI - Enzyme treatment of exocrine pancreatic insufficiency in chronic pancreatitis. AB - Exocrine pancreatic insufficiency combined with pancreatic pain and endocrine pancreatic insufficiency are the leading symptoms of chronic pancreatitis. Due to the large functional reserve capacity of the gland, decompensation, i.e. steatorrhea, does not occur before lipase excretion is reduced to < or = 10% of normal. Pancreatic enzyme substitution is indicated when fecal fat excretion exceeds a critical value (normally > 15 g/day) and/or when weight loss is present. A number of studies have dealt with the problems of gastric acid inactivation of pancreatic enzyme preparations as well as their gastric emptying nonsimultaneously with the food. For the present, it is recommended that pancreatic enzyme substitution in patients with proven exocrine pancreatic insufficiency and normal gastric acid secretion be given in multiunit, acid protected dosages. In patients with gastric hyposecretion and in those who underwent partial or total gastrectomy, enzyme substitution should be administered as granules to enable mixing and simultaneous transport of enzymes with the chyme. The ultimate aim of further scientific and clinical research remains the total abolishment of pancreatic steatorrhea. PMID- 8224570 TI - Acute pancreatitis: when is enzyme treatment indicated? AB - Discussion over the role of enzyme therapy in patients with acute pancreatitis is based upon theoretical considerations rather than hard data. In this respect, it differs little from most other treatments used in the management of acute pancreatitis, although, in the case of enzyme treatment, the lack of data results largely from a dearth of experimentation to prove its efficacy rather than from equivocal or negative results. PMID- 8224571 TI - Indication for pancreatic enzyme treatment in non-pancreatic digestive diseases. AB - Several studies have shown that exocrine pancreatic function may be impaired in various non-pancreatic digestive diseases. The impairment, however, is usually mild to moderate and generally does not cause malabsorption. In some patients, however, pancreatic insufficiency is severe enough to cause malabsorption, which can be effectively corrected by oral pancreatic extracts. PMID- 8224572 TI - Enzyme treatment after gastrointestinal surgery. AB - After gastrointestinal surgery, patients often suffer from maldigestion. The extent of this maldigestion syndrome depends on the type of surgical procedure performed. After total pancreatectomy, subtotal left resection, resection for chronic pancreatitis. Whipple operation with ductal occlusion, and total gastrectomy, patients need obligatory enzyme treatment. After partial pancreatectomy without duct occlusion or partial gastrectomy, enzyme treatment should be initiated when exocrine pancreatic insufficiency occurs. PMID- 8224573 TI - Otoacoustic emissions recorded at rapid stimulus rates. AB - Otoacoustic emissions were evoked by clicks presented according to maximum length sequences. The analysis disentangled the overlapping responses that occur when stimuli are presented at intervals shorter than the duration of the response. Two types of sequence were used: a "monopolar" sequence in which clicks were present or absent, and a "bipolar" sequence in which both condensation and rarefaction clicks occurred. Emissions can be recorded at stimulus rates of up to 1000/sec although the amplitude of the response decreases at these rapid rates. This decrease in amplitude requires a compensatory increase in the number of stimuli to maintain the same signal-to-noise ratio. Optimal efficiency can be attained at rates between 200 and 500/sec. At these rates the emissions can be recorded in about one half the time required for conventional averaging at regular rates of 50/sec. PMID- 8224574 TI - Confidence intervals for hearing loss allocation estimates. AB - A method for estimating confidence intervals for hearing loss allocation estimates is developed and demonstrated. Based on the ISO-1999 model for the combination of noise-induced permanent threshold shift (NIPTS) and age-related permanent threshold shift (ARPTS), the probabilities of different pairs of NIPTS and ARPTS values can be represented by a three-dimensional joint probability density function (JPDF). When the JPDF is intersected by equal-probability planes, the resulting ellipses define the confidence intervals. This method makes conservative assumptions that will usually overestimate the width of the confidence interval. PMID- 8224575 TI - Factors affecting laterality of standard threshold shift in occupational hearing conservation programs. AB - Although noise-induced hearing loss is generally characterized as bilaterally symmetrical, 80% of audiometric shifts meeting Occupational Safety and Health Administration (OSHA) criteria were unilateral in a large industrial sample. The primary factor related to unilateral OSHA shifts was asymmetric baseline hearing level. Better ears at baseline audiogram were more likely to demonstrate OSHA shifts than poorer ears. When hearing levels were symmetric at baseline, left ears were more likely to demonstrate OSHA shifts. Although OSHA shifts were primarily unilateral, cumulative effects of noise exposure were judged to be bilaterally symmetrical due to larger hearing shifts occurring in better ears of subjects with preexisting hearing asymmetries. Recovery from temporary threshold shift in second ears tested was judged not to be a significant determinant for high prevalence of unilateral shifts. Many factors likely interact to confound interpretive value of OSHA shift laterality in occupationally noise-exposed populations. This holds important implications for hearing conservation program professionals considering hearing shift laterality as a factor in shift etiology. PMID- 8224576 TI - Acoustical properties of speech produced in noise presented through supra-aural earphones. AB - This study determined the acoustical properties of speech known as Lombard Speech produced in background noise. Tape recordings were made for ten normally hearing adults (5 women, 5 men) reading connected speech (131 word passage "My Grandfather") at their most comfortable level in quiet and in wideband, traffic, and multitalker noise delivered through earphones at 70 and 90 dB SPL. Spectral analysis of the recordings revealed that, compared with speech in quiet, Lombard speech was characterized by: (1) an increase in overall SPL; (2) smaller vocal pitch shifts for female than male subjects; (3) shifts in spectral distributions of speech energy; and (4) the same spectral slope above 630 Hz regardless of subject gender, noise level, or noise type. Overall, the results of this study do not provide support for the theory that acoustical properties of Lombard speech are identical with loud speech produced in quiet. PMID- 8224577 TI - The effects of auditory feedback from the nucleus cochlear implant on the vowel formant frequencies produced by children and adults. AB - Cochlear implants provide an auditory signal with which profoundly deaf users may monitor their own speech production. The vowel production of two adults and three children who used the Nucleus multiple-electrode cochlear implant was examined to assess the effect of altered auditory feedback. Productions of words were recorded under conditions where the talkers received auditory feedback (speech processor turned on) and where no auditory feedback was provided (speech processor turned off). Data were collected over 3 days at weekly intervals. First and second formant frequencies were measured and the data were analysed to assess significant differences between auditory feedback conditions, vowel context, and data collection points. Overall, the results varied across talkers, across the data collection days, and depended on the consonant environment of the vowel. However, two effects of auditory feedback were noted. First, there was a generalized shift in first formant frequencies between the processor on and processor off conditions across three of the five subjects, but the shift differed in direction for each subject. Second, for three of the five talkers, the two front vowels /epsilon/ and /i/ were more neutralised in the absence of auditory feedback. However, this effect was less pronounced than that noted by previous studies. PMID- 8224578 TI - Perceptual performance of subjects with cochlear implants using the Spectral Maxima Sound Processor (SMSP) and the Mini Speech Processor (MSP). AB - A portable sound processor has been developed at the University of Melbourne for use with the 22-electrode cochlear implant manufactured by Cochlear Pty Limited. In this report, the performance of the Spectral Maxima Sound Processor (SMSP) is compared with the Mini Speech Processor (MSP) presently used clinically with this implant. Results of tests with 5 subjects demonstrate that the performance of the SMSP significantly exceeds that of the MSP for recognizing vowels and consonants in closed-set confusion studies and open-set monosyllabic word tests, although recognition of speaker identity and intonation patterns is similar for both processors. Information transmission analysis of phoneme features confirms that most features, such as vowel format frequencies and consonant manner and place of articulation, are better conveyed with the SMSP. Possible reasons for its improved performance are discussed. PMID- 8224579 TI - Measuring the effects of head and neck support systems on signals transmitted to the ear. AB - Some individuals with motor control problems require head and/or neck support systems to achieve proper positioning in their wheelchairs. Signal transmission to a KEMAR positioned in a wheelchair with three commonly used support systems was evaluated with a probe microphone system. The three systems created different patterns of shadow and baffle effects. Audiologists are advised to evaluate the acoustic impact of the specific support systems used by their patients. PMID- 8224580 TI - [Immunophysiologic mechanisms of hypoxia]. PMID- 8224581 TI - [Age- and sex-related characteristics of the pituitary-adrenal and insular systems in healthy people]. PMID- 8224582 TI - [Taste perception and glycemic reaction in disorders of glucose tolerance]. PMID- 8224583 TI - [Regulation of carbohydrate metabolism in people living in the North]. PMID- 8224584 TI - [Levels of Ca, Na, K, Mg in plasma and blood serum ultrafiltrate in children with rickets]. PMID- 8224585 TI - [Migrating rhythmic phase in motor activity of the gastrointestinal tract in the process of digestion in healthy men]. PMID- 8224586 TI - [Features of mental working capacity of sluggish and active children in grades I IV, living in extreme climatic conditions]. PMID- 8224587 TI - [Electromyographic criteria for the human response to small and average doses of radiation]. PMID- 8224588 TI - [Basic structural and functional diagram of a clone reflex network]. PMID- 8224589 TI - [Development of walking]. PMID- 8224590 TI - [A method of predicting human thermal adaptation]. PMID- 8224591 TI - [Simple motor reaction time and peripheral neurosensory hypoacusis]. PMID- 8224592 TI - [Sex differences in EEG spectral parameters in 6-7 year old children]. PMID- 8224593 TI - [Supraspinal mechanisms of regulation of muscle tonus and their correction in patients with Parkinsonism using recorded omega-potentials]. PMID- 8224594 TI - [Neurophysiologic correlates of rehabilitation of central motor disorders using spinal fluid transfusion]. PMID- 8224595 TI - [Trace hyperpolarization of motor neurons and h-reflex rehabilitation curve of slow and fast muscles]. PMID- 8224596 TI - [Psychophysiologic mechanisms of attention deficit in children of various ages with learning difficulties]. PMID- 8224597 TI - [Dynamic properties of body motion and effectiveness of motor control]. PMID- 8224598 TI - [Rate of pulse wave propagation with changing limb length and regional arterial pressure in subjects of varying age]. PMID- 8224599 TI - [Functional correlation between psychophysiologic parameters of autonomic regulation in athletes of various sports]. PMID- 8224600 TI - [Psychophysiologic criteria in predicting the working efficiency of power plant operators at their admission to the training center]. PMID- 8224601 TI - [Biological age and physical working capacity based on longitudinal observations in factory and office workers in the ship-building industry]. PMID- 8224602 TI - [Analysis of individual differences of psychological reactions to combined hypoxic exposure]. PMID- 8224603 TI - Scientists must educate the public that NIH is the principal federal agency that supports biomedical research. PMID- 8224604 TI - A statistical model validating triage for the peer review process: keeping the competitive applications in the review pipeline. AB - Triage of grant application at the National Institutes of Health (NIH) is a process whereby an initial screening of applications by a scientific peer review group eliminates applications that are not competitive for awards. The process of application triage has been limited to those applications submitted to the NIH in response to an RFA (Request for Applications). A hypergeometric model was developed to determine the extent to which five, six, seven, or eight member triage teams or subsets of 12-to-20 member full committees could provide a statistically defensible triage decision. Although the intent of triage is to remove from review those applications that are noncompetitive, the model was weighted in favor of the applicant to minimize the likelihood that highly competitive applications would be eliminated. Within the assumptions and rules developed, it was determined that there was little likelihood that the latter would occur. For example, in the worst case scenario, the greatest probability that a highly competitive application would be knocked out of competition is P < or = 0.014 in the case of a five-member triage subset of a 20-member committee. Using the latter case, the model was tested on a set of 73 applications that were submitted to the National Cancer Institute for action at the February 1993 National Cancer Advisory Board. The model selected for triage required that each application be assigned to five reviewers, that each reviewer be blinded to the review assignments of the other reviewers, and that four noncompetitive votes be registered to triage out an application. Each of 19 applications received four to five noncompetitive votes, and were triaged out of the review process. The remaining 54 applications were then reviewed according to the usual NIH review process. Four of the applications received three noncompetitive triage votes each and were either rated as not recommended for further consideration (NRF, n = 2)) or received priority scores > or = 250 (n = 2) (The smaller the priority score the better the technical merit). Thirteen of the 53 applications received two noncompetitive votes. Of the latter, two were not recommended for further consideration and the remaining 11 received priority scores between in excess of 200. The distribution of competitive applications was such that funding was limited to those applications with priority scores of less than 190. Thus, the data suggest that the conservative model is valid such that the likelihood of eliminating a highly competitive application from consideration for funding is remotely small.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8224605 TI - Extracellular matrix 5: adhesive interactions in early mammalian embryogenesis, implantation, and placentation. AB - Normal morphogenesis and differentiation depend heavily on the coordination of cell-cell and cell-extracellular matrix interactions. During early mammalian development, the first cell lineages to be established are extraembryonic (trophoblast and extraembryonic endoderm), which are essential for satisfying the nutritional requirements of the developing embryo. This review emphasizes the importance of the cadherin family of cell-cell adhesion molecules and the integrin family of extracellular matrix receptors in mediating interactions between cells and their environment during early development. The review first discusses the critical role of cell-cell interactions in fertilization and early lineage decisions that occur during pre- and peri-implantation development in the mouse, using the calcium-dependent cell adhesion molecule E-cadherin as the primary example. The remainder of the review discusses the importance of cell-ECM interactions in the further morphogenesis and differentiation of the newly segregated lineages. The critical roles of integrins in differentiation, migration, and invasion of trophoblast in both mouse and human are emphasized. PMID- 8224606 TI - Concepts and principles of glycobiology. AB - In biological systems oligosaccharides are normally conjugated to proteins or lipids. The heterogeneity and branching of oligosaccharides allow glycoconjugates to display a further level of structural and functional diversity compared with linear proteins and nucleic acids or with lipids. This review summarizes some general principles that are emerging from the new field of glycobiology which, by addressing the molecular interactions of glycoconjugates in biological systems, spans the classical physicochemical, biological, and biochemical sciences. We discuss the genesis of glycoforms, the functional roles for glycosylation, and some general aspects of structure/function relationships with reference to N glycosylated animal glycoproteins including the enzymes ribonuclease and tissue plasminogen activator, IgG, the family of C-type lectins, and receptor ligands. PMID- 8224607 TI - Bacterial ice nucleation: significance and molecular basis. AB - Several bacterial species are able to catalyze ice formation at temperatures as warm as -2 degrees C. These microorganisms efficiently catalyze ice formation at temperatures much higher than most organic or inorganic substances. Because of their ubiquity on the surfaces of frost-sensitive plants, they are responsible for initiating ice formation, which results in frost injury. The high temperature of ice catalysis conferred by bacterial ice nuclei makes them useful in ice nucleation-limited processes such as artificial snow production, the freezing of some food products, and possibly in future whether modification schemes. The rarity of other ice nuclei active at high subfreezing temperature, and the ease and sensitivity with which ice nuclei can be quantified, have made the use of a promoterless bacterial ice nucleation gene valuable as a reporter of transcription. Target genes to which this promoter is fused can be used in cells in natural habitats. Warm-temperature ice nucleation sites have also been extensively studied at a molecular level. Nucleation sites active at high temperatures (above -5 degrees C) are probably composed of bacterial ice nucleation protein molecules that form functionally aligned aggregates. Models of ice nucleation proteins predict that they form a planar array of hydrogen binding groups that closely complement that of an ice crystal face. Moreover, interdigitation of these molecules may produce a large contiguous template for ice formation. PMID- 8224608 TI - Noncholinergic functions of cholinesterases. AB - Cholinesterases (acetylcholinesterase and butyrylcholinesterase) exhibit additional catalytic activities apart from their well-known action in hydrolyzing choline esters. An amine-sensitive aryl acylamidase activity is exhibited by both acetyl- and butyrylcholinesterases. A metallocarboxypeptidase-like activity is found associated with both acetyl- and butyrylcholinesterases. The peptidase activity exhibited by butyrylcholinesterase was located in a 50-kDa COOH-terminal fragment. Acetylcholinesterase is implicated in noncholinergic functions in the substantia nigra. A relationship between tumorigenesis, cell differentiation, and cholinesterases has been speculated. The sequence similarities between different esterases, lipases, thyroglobulin, cell adhesion proteins, and cholinesterases would make it appear that cholinesterases are capable of exhibiting more than one biological activity and their functions are wider than what is hitherto known. PMID- 8224609 TI - Proatherosclerotic events: pathobiochemical changes occurring in the arterial wall before monocyte migration. AB - The inception of experimentally induced atherogenesis is marked by subtle biochemical and ultrastructural changes of the arterial intima, modifications that precede the monocytes migration within the vessel wall. Long before any recognizable endothelial denudation, hypercholesterolemia induces a sequence of prelesional events that start with the enhanced transport of excess plasma lipoproteins especially by transcytosis, followed by intima accumulation of chemically (oxidatively) modified and reassembled lipoproteins (MRLp). Exposure of endothelial cells concomitantly to hypercholesterolemia on the luminal side, and to the MRLp cytotoxic effects on the abluminal side, generates endothelial dysfunctions with altered biosynthetic activities. The latter are manifested initially by the hyperplasia of basal lamina and its disjunction from endothelium, and by the proliferation and reorganization of a modified extracellular matrix that traps MRLp. The cytotoxic effects of MRLp contribute to the endothelial production of chemoattractants and adhesion molecules that are instrumental in monocyte recruitment and migration in the subendothelium, where activated and differentiated as macrophages, they avidly ingest MRLp and their complexes to form foam cells. The latter are the hallmark of fatty streaks that marks the transition from the prelesional to the lesional stage of atherogenesis. Understanding the earliest biochemical alterations of the artery wall is a prerequisite for designing more successful means of prevention and treatment of atherosclerosis. PMID- 8224610 TI - Lysophosphatidic acid mimics serum-induced sensitization of cyclic AMP accumulation. AB - Pretreatment of 1321N1 human astrocytoma cells with serum induces a pronounced increase in subsequent stimulation by forskolin and other agents of intracellular cyclic AMP accumulation, a phenomenon referred to as sensitization (Mol. Pharmacol. 39, 399-406, 1991). Pretreatment of these cells with lysophosphatidic acid induced sensitization to a similar extent as that with serum (approximately fivefold for forskolin stimulation and twofold for isoproterenol and prostaglandin E1 stimulation), with half-maximal effects at approximately 30 nM lysophosphatidic acid. Phosphatidic acid was effective but less potent whereas other lipids were ineffective. Sensitization by serum and by lysophosphatidic acid were almost completely inhibited by pertussis toxin pretreatment and partially inhibited by prolonged phorbol ester exposure to induce protein kinase C down-regulation. Among nine cell lines tested, those that exhibited sensitization with serum showed comparable sensitization with lysophosphatidic acid. The effects of both lysophosphatidic acid and serum were markedly inhibited by treatment with phospholipase B but only minimally altered with phospholipases A2, D, and C. Exposure of cells to phospholipase C alone induced approximately threefold sensitization, but both serum and lysophosphatidic acid were able to induce further three- to fourfold sensitization above that induced by phospholipase C alone. In contrast, the effects of serum and lysophosphatidic acid were not additive with each other. Together these results suggest that lysophosphatidic acid or a closely related compound present in serum is the factor responsible for sensitization of the cyclic AMP pathway. PMID- 8224611 TI - Glycogen synthesis in the astrocyte: from glycogenin to proglycogen to glycogen. AB - The astrocyte of the newborn rat brain has proven to be a versatile system in which to study glycogen biogenesis. We have taken advantage of the rapid stimulation of glycogen synthesis that occurs when glucose is fed to astrocytes, and the marked limitation on this synthesis that occurs in astrocytes previously exposed to ammonium ions. These observations have been related to our earlier reports of the initiation of glycogen synthesis on a protein primer, glycogenin, and the discovery of a low-molecular-weight form of glycogen, proglycogen. The following conclusions have been drawn: 1) In the ammonia-treated astrocytes starved of glucose, free glycogenin is present. 2) When these astrocytes are fed with glucose, proglycogen is synthesized from the glycogenin primer by a glycogen synthase-like UDPglucose transglucosylase activity (proglycogen synthase) distinct from the well-recognized glycogen synthase, and synthesis stops at this point. 3) Proglycogen is the precursor of macromolecular glycogen, which is synthesized from proglycogen by glycogen synthase when glucose is fed to untreated astrocytes, accounting for the much greater accumulation of total glycogen. 4) The stimulus to proglycogen and macroglycogen synthesis that occurs on feeding glucose to untreated or ammonia-treated astrocytes is the result of the activation of proglycogen synthase, not of glycogen synthase. 5) Therefore, in the synthesis of macromolecular glycogen from glycogenin via proglycogen, the step between glycogenin and proglycogen is rate-limiting. 6) The discovery of additional potential control points in glycogen synthesis, now emerging, may assist the identification of so-far-unexplained aberrations of glycogen metabolism. PMID- 8224612 TI - A potential pathophysiologic role for alpha 2 beta 1 integrin in human eye diseases involving vitreoretinal traction. AB - Cell-mediated contraction of tissues containing fibrillar collagens can lead to organ compromise and loss of function. The same process that is biologically advantageous during the contraction phase of wound healing can be subverted in diseases such as hepatic cirrhosis, pulmonary fibrosis, and scleroderma, although the cellular and molecular mechanism of matrix tissue contraction is difficult to study in such chronic diseases. However, certain human eye diseases that result in tractional detachment of the retina and loss of vision are characterized by acute cell-mediated contraction of collagenous tissue in the vitreous cavity. In this study, we demonstrate that human cells can contract vitreous, a complex biological gel containing type II collagen, in vitro. This cell-mediated contraction can be blocked by antibodies and peptides that antagonize the function of alpha 2 beta 1 integrin, and the potential for contraction can be conferred upon noncontracting cells by stable transfection of cells with alpha 2 cDNA. We also show that this contractile process, if focally resisted, can result in remodeling vitreous from a gel to a structure that resembles a planar membrane, and that substantial isometric forces can be measured across this tissue. We propose that in diseases such as proliferative diabetic retinopathy and proliferative vitreoretinopathy, alpha 2 beta 1 integrin-mediated contraction of the vitreous and tension at the site of vitreoretinal attachments contribute to the terminal event of tractional retinal detachment. By extension, we propose that alpha 2 beta 1 integrin is a centrally important molecule in human diseases characterized by remodeling and contraction of collagenous tissue (i.e., fibrocontractive diseases). PMID- 8224613 TI - Retinoblastoma and p53 tumor suppressor genes in human hepatoma cell lines. AB - We analyzed the status of retinoblastoma and p53 genes in 10 human hepatoma cell lines. Polyclonal anti-peptide antibodies generated against peptides homologous to COOH-terminal and leucine-zipper domains of the retinoblastoma protein allowed us to identify two cell lines (Hep 3B and FOCUS) with abnormal expression. The same cell lines have both lacked p53 expression. In contrast to the retinoblastoma gene, the expression of the p53 gene was abnormal in six additional cell lines. Indeed, only the Hep G2 hepatoblastoma cell line (and its derivative Hep G2/2215) appeared to have normal p53 and retinoblastoma gene expression. Our studies indicate that p53 abnormalities are common but retinoblastoma gene aberrations are rare in human hepatoma cell lines. PMID- 8224614 TI - Rauhsche Nase--then and now. PMID- 8224615 TI - Comparison of portal pressure with intravascular esophageal variceal pressure (IEVP) directly measured with a flexible indwelling needle. AB - Intravascular esophageal variceal pressure (IEVP) was measured using a flexible indwelling needle and compared with wedged hepatic venous pressure (WHVP) in 38 patients with liver cirrhosis. There was a high correlation between IEVP and WHVP; the former was lower than the latter, indicating the presence of a pressure gradient. Bleeders with a history of esophageal variceal rupture showed higher IEVP value than non-bleeders without a history of rupture, even though no difference in WHVP was observed. IEVP in the post-endoscopic injection sclerotherapy (EIS) group, even in those with endoscopic findings of large varices, was lower than that in the untreated groups. These results suggest that IEVP is a factor in the hemodynamics of patients with portal hypertension and that its measurement may be helpful in elucidating the pathophysiology of esophageal varices. PMID- 8224616 TI - The effect of acid secretagogues on mucin synthesis using primary monolayer culture of the guinea pig gastric mucous cells. AB - Mucin plays a principal role in protecting the gastric mucosa against injury. We investigated the effect of acid secretagogues on mucin synthesis using a primary gastric mucous cell monolayer culture system of guinea pig. Significant increases in mucin synthesis were observed in response to the secretagogues pentagastrin (10(-8)M, 10(-7)M) and carbachol (10(-4)M, 10(-3)M), but not to histamine. After pretreatment with indomethacin (10(-5)M), 10(-8)M pentagastrin significantly increased mucin synthesis to 125.6 +/- 3.9%, but carbachol did not. Prostaglandin E2 release into the culture medium was significantly increased by 10(-4)M carbachol to 118.0 +/- 5.9%, but there was no change after application of pentagastrin. These findings suggest that pentagastrin and carbachol may act directly on mucous cells, and that part of the mucin synthesis-promoting action of carbachol is mediated by prostaglandins. There were no changes in intracellular cAMP concentration after the addition of these acid secretagogues. However, calcium ionophore (A23187) produced an increase in mucin synthesis, suggesting a Ca++ involvement in mucin synthesis. No differences were found in the sugar side chain structures of newly synthesized mucin glycoprotein as a result of exposure to acid secretagogues by histochemical or biochemical methods using lectins. PMID- 8224617 TI - Plasma endothelin-1 concentrations are elevated in acute hepatitis and liver cirrhosis but not in chronic hepatitis. AB - We measured plasma endothelin-1 (ET-1) concentrations in 20 healthy controls and 63 patients with liver diseases including 9 cases of acute hepatitis (AH), 14 cases of chronic hepatitis (CH), 24 cases of liver cirrhosis (LC), 11 cases of hepatocellular carcinoma with LC (HCC), 3 of primary biliary cirrhosis and 2 of idiopathic portal hypertension. ET-1 levels in AH (5.07 +/- 2.54 pg/ml, mean +/- SD), LC (3.71 +/- 1.17) and HCC (3.08 +/- 0.93) were significantly higher than those in healthy controls (2.18 +/- 0.37). ET-1 levels in AH, LC and HCC were also significantly higher than those in CH (2.05 +/- 0.61). ET-1 levels showed negative correlations with serum albumin levels and Ch-Ease activities, and positive correlations with serum bilirubin levels, AST and ALT activities. However, there was no correlation between plasma ET-1 concentrations and concentrations of serum thrombomodulin which is known to be a marker of injured vascular endothelial cells. In cirrhotic patients, ET-1 levels were significantly influenced by the presence of ascites. The results of the present study suggest that plasma ET-1 concentrations may be a useful clinical indicator for use in the follow-up of patients with chronic liver diseases, e.g., progression from CH to LC, and change in grade of portal hypertension and decompensation in LC. PMID- 8224618 TI - Alpha-interferon enhances gamma-interferon production of peripheral blood mononuclear cells pre-activated with phytohemagglutinin. AB - The question of whether the state of T cell activation is responsible for gamma interferon production in response to alpha-interferon was determined. When peripheral blood mononuclear cells were pre-incubated with phytohemagglutinin for 3 days, their gamma-interferon production was found to be augmented by alpha interferon, accompanied by an increased proportion of DR-positive T cells. This did not occur with fresh cells. The effect was dose-dependent and inhibited by anti-alpha-interferon antibody. Therefore, alpha-interferon was considered to enhance the production of gamma-interferon of pre-activated peripheral blood mononuclear cells but not of resting ones. In conclusion, alpha-interferon augmented the production of gamma-interferon of peripheral blood mononuclear cells only when they had already been activated, suggesting a role of alpha interferon as a modulator of the cellular immune response. PMID- 8224619 TI - Serum pancreatic phospholipase A2 and prophospholipase A2 in acute pancreatitis and after endoscopic retrograde pancreatography. AB - Serum immunoreactive pancreatic phospholipase A2 (IP-PLA2) levels and the proportion of active pancreatic PLA2 in the serum IP-PLA2 were determined by radioimmunoassay with an antibody directed against active human PLA2. The subjects of this study included eight patients who underwent endoscopic retrograde pancreatography (ERP), nine patients with acute pancreatitis, and six healthy controls. The serum IP-PLA2 levels were elevated after ERP and during acute pancreatitis. The amount of active pancreatic PLA2 in the serum was determined after its separation from pancreatic prophospholipase A2 using reverse phase high-performance liquid chromatography (HPLC). The serum IP-PLA2 was separated into two peaks on reverse-phase HPLC. The one which eluted faster contained the PLA2 activity; the other peak did not. The latter IP-PLA2 peak consisted of pancreatic prophospholipase A2 as judged by HPLC analysis and PLA2 activity determination of the products after treatment with trypsin. The proportion of active pancreatic PLA2 in the serum IP-PLA2 of patients after ERP (13.9 +/- 0.5%) increased slightly compared with that in fasting, healthy controls (8.0 +/- 1.1%). For those with acute pancreatitis, the proportion of active pancreatic PLA2 within 48 hours of hospital admission increased more markedly (44.0 +/- 5.7%) than that after ERP. These findings demonstrate that the proportion of active pancreatic PLA2 in the serum IP-PLA2 markedly increases during the early stage of acute pancreatitis, and that an ERP-induced rise in the intraductal pressure leads to the leakage of pancreatic PLA2 into the circulation, but not to a marked activation of the leaked enzyme. PMID- 8224620 TI - Protease-antiprotease levels and whole-blood chemiluminescence in acute peritonitis. AB - Whole-blood chemiluminescence and levels of leukocyte proteases and plasma protease inhibitors were studied in 43 patients with acute, generalized peritonitis. An almost three-fold increase in whole-blood chemiluminescence was found in acute peritonitis, which may indicate activation or "priming" of the leukocytes by blood-borne factors. High levels of leukocyte elastase and neutrophil proteinase 4(3) were found in plasma and peritoneal exudate. Patients with sepsis had higher plasma levels of both proteases than other patients. Large variations in the plasma levels among patients decreased their sensitivity as markers of infectious complications during the postoperative period. The plasma levels of the protease inhibitors followed three different patterns of reaction. The acute phase proteins alpha 1-proteinase inhibitor and C1-inactivator, increased during the first week of disease, to normalise later in its course. alpha 2-macroglobulin, antithrombin III and alpha 2-antiplasmin were all decreased from onset and normalised later in the course, while secretory leukocyte protease inhibitor showed a slow decrease throughout the course of disease. In peritonitis exudate, the levels of the main protease inhibitors, alpha 1-Proteinase Inhibitor and alpha 2-Macroglobulin, were decreased, probably due to complexation and subsequent elimination, as a part of the defense against liberated leukocyte proteases. The immunoreactive and especially functional levels of the protease inhibitors alpha 2-Antiplasmin, Antithrombin III and C1 Inactivator were also decreased in the exudate, indicating an increased turn-over of these proteins through activation of the cascade systems and/or break-down by leukocyte proteases. In contrast to the other inhibitors, secretory leukocyte protease inhibitor showed higher levels in exudate than in plasma, and unexpectedly high exudate/plasma-quotients were seen in cases with colonic perforations. Degradation of complement factor 3 (C3) and decreased "opsonic capacity" were found in exudate. The "opsonic capacity" could be correlated to the levels of leukocyte proteases in the exudate, which indicates that degradation of complement factor 3 may have been at least partly due to the action of leukocyte proteases. Further depletion of complement factors in exudates of long-standing peritonitis or abscesses may create a vicious circle of deficient opsonisation and clearance of bacteria, as earlier reported for chronic pleural exudates. PMID- 8224621 TI - Retroperitoneal fibrosis associated with scirrhous gastric cancer. AB - A case of retroperitoneal fibrosis associated with scirrhous gastric cancer is reported. A sixty two-year-old Japanese female was admitted because of acute renal failure. The patient's serum creatinine level showed 3.2 mg/dl while the blood urea nitrogen level was 23 mg/dl. An ultrasound study of the upper abdomen revealed bilateral hydronephrosis. Drip infusion pyelography revealed a dilated right renal pelvis without ureteral obstruction. The left kidney was not opacified, suggesting a functional disorder. Gastrography and gastrofiberscopy revealed scirrhous gastric cancer. Signet ring cell carcinoma was later demonstrated histologically by biopsy specimens. CT demonstrated a prominent thickening of the gastric wall and hydronephrosis, although no prevertebral soft tissue masses were observed. A total gastrectomy was performed with failure to surgically decompress the ureters because fibrous plaque had firmly enveloped the retroperitoneal structures. Biopsy specimens of the retroperitoneum revealed an invasion of the tumor cells and prominent fibrosis. As an etiology of renal failure, ureteral stenosis resulting from secondary retroperitoneal fibrosis was also considered. PMID- 8224622 TI - Ileal obstruction due to metastatic liposarcoma: a case report. AB - Liposarcoma in the mesentery or small intestine is rare. Intestinal obstruction due to liposarcoma is particularly rare. We report here the case of a 33-year-old man with intestinal obstruction due to metastasis of a liposarcoma to the ileum. Although he died due to multiple metastases, it should be emphasized that he recovered from intestinal obstruction by resecting the tumor in the ileum. The pertinent literature is also briefly reviewed. PMID- 8224623 TI - A case of polypoid cavernous hemangioma of the sigmoid colon excised by colonoscopic polypectomy. AB - A 28-year-old Japanese man was admitted to our institute because of sharp anal pain and a mass that prolapsed from the anus after defecation. After spinal anesthesia for emergency operation, the mass spontaneously withdrew into the anus and the pain disappeared. Surgery was postponed. Barium enema and colonoscopy revealed a pedunculated polyp in the sigmoid colon, which was removed by snare polypectomy with electrocautery. The resected polyp was granular and reddish, and measured 33 x 22 x 14 mm. Histological examination of the polyp revealed a cavernous hemangioma. Hemangioma of the colon is rare; only 52 cases have been reported in Japan. Of these patients, eight underwent endoscopic polypectomy. The present lesion is the largest thus treated that has been documented. Endoscopic polypectomy seems to be a safe and preferable procedure for the diagnosis and treatment of small, solitary polypoid hemangiomas of the colon. PMID- 8224625 TI - A successful case of intraarterial prednisolone injection therapy in severe ulcerative colitis unresponsive to an intensive intravenous regimen. AB - A successful case of intraarterial prednisolone injection therapy in severe ulcerative colitis unresponsive to conventional therapy, including an intensive intravenous regimen, is presented. After the direct administration of prednisolone into the mesenteric arteries, the patient's conditions markedly improved, and he succeeded in overcoming the acute stage and entered remission. Accordingly, he safely underwent elective surgery which was finally required because of the inability to withdraw prednisolone therapy. It is well worthwhile to consider this therapy in the last resort of medical therapy for ulcerative colitis. PMID- 8224624 TI - A case of perianal Paget's disease associated with a sigmoid colon carcinoma. AB - A case of perianal invasive Paget's disease associated with a sigmoid colon carcinoma is presented. The chief complaint was perianal irritation for a year. Histologic examination yielded a correct diagnosis and abdominosacral resection with wide excision of the cutaneous component was performed. Histology of the resected specimen revealed the Paget's disease to be invasive of the dermis, the sigmoid colon carcinoma to be in Dukes' A stage and the two lesions to be different. The patient has been disease-free for more than 5 years after the operation. PMID- 8224626 TI - Liver sarcoidosis showing low-density intrahepatic septa on postcontrast computed tomography. AB - The authors present a patient with massive liver sarcoidosis, showing only slight hepatomegaly on precontrast computed tomogram and multiple low-density intrahepatic septa on postcontrast computed tomogram. Sarcoidosis frequently involves the liver, but rarely shows abnormal findings other than hepatomegaly on computed tomograms. Only a few cases have demonstrated a low density intrahepatic area on computed tomograms. In this report, we present a patient with massive liver and pulmonary sarcoidosis, showing low density intrahepatic septa and pulmonary fibrosis on computed tomograms. PMID- 8224627 TI - A case of chronic pancreatitis complicated by massive pericardial and right pleural effusion. AB - A 42-year-old man was admitted complaining of dyspnea. Chest X-ray showed an increase in cardiac size, and echocardiography revealed a large volume of pericardial effusion. Pancreatic enzyme levels were elevated in both serum and pericardial effusion. Computed tomography and endoscopic retrograde pancreatography demonstrated a fistula connecting a pancreatic pseudocyst with the pericardium and the right pleural cavity. Massive pericardial and right pleural effusion is an extremely rare complication of chronic pancreatitis. In this case, computed tomography and endoscopic retrograde pancreatography were useful for diagnosing the fistula. PMID- 8224628 TI - Expression of transforming growth factor-beta receptor type II gene in gastric cancer cell lines. PMID- 8224629 TI - Serum concentration of macrophage colony stimulating factor (M-CSF) in patients with inflammatory bowel disease. PMID- 8224630 TI - Cisplatin enhances the metastatic potential of human colon carcinoma. PMID- 8224631 TI - Immunology of the intestinal tract. AB - The last half century has witnessed the birth, infancy, and early adolescence of a new field, intestinal immunology, with an explosion of knowledge about the role of the intestinal immune system in normal intestinal physiology and disease. This field is still in its rapid growth phase. A major spin-off of such growth will be the development of new approaches for the diagnosis and treatment of disease. For example, studies of oral tolerance have led to new insights into possible mechanisms of autoimmunity and ultimately could lead to new treatments for autoimmune disease. The discovery that Peyer's patches are a major inductive site for initiation of the mucosal immune response, and IgA responses in particular, and that antigen enters these sites via the M cell, forms one current basis for the development of new mucosal vaccines and vaccine delivery systems. Studies of immunogenetics are leading to new insights into the molecular basis of diseases such as celiac disease, IBD, and hepatic disorders. Studies of cytokines and the role they play in acute and chronic inflammation are leading to new approaches for the treatment of intestinal inflammatory diseases. The field of intestinal immunology is now well on its way through the turbulent "teenage" years. It is true maturation over the next several decades will test and witness the evolution and validity of concepts formed over the past 50 years. Certainly, tremendous growth and change will come with the increasing application of molecular tools, and the enthusiasm and the fresh insights brought by the next generation of young investigators, as they begin to further unravel the intricacies of the intestinal immune system and its role in health and disease. PMID- 8224632 TI - Pyxigraphic sampling to enumerate methanogens and anaerobes in the right colon of healthy humans. AB - BACKGROUND: A major limitation in studying the proximal human colonic flora is the lack of suitable sampling methods. The aim of this study was (1) to describe a prototype technique, pyxigraphy, which uses swallowed capsules containing a mechanism allowing the remote control of sampling the gastrointestinal tract contents, and (2) to use this sampling method to examine the distribution of methanogens in the colon of methane (CH4) excretors and non-CH4 excretors. METHODS: In six CH4 excretors and four non-CH4 excretors, samples of the right colonic contents were obtained by means of the pyxigraphic sampling method. Methanogens and total anaerobes were enumerated in both the right colonic and fecal contents. RESULTS: In CH4 excretors, the concentration of methanogens was higher in the feces than in the right colonic contents, representing 12% and 0.003%, respectively, of the total anacrobes (P < 0.02). In non-CH4 excretors, no difference was observed, methanogens representing < 0.003% of the total anaerobes in both the right colonic and fecal contents. CONCLUSIONS: Pyxigraphy is a noninvasive, simple, and safe sampling method that allows to study the microbial populations of the proximal colon. The results obtained showed that methanogens preferentially colonize the distal part of the colon in CH4 excretors. PMID- 8224633 TI - Uptake of riboflavin across the brush border membrane of rat intestine: regulation by dietary vitamin levels. AB - BACKGROUND: Possible regulation of riboflavin (RF) intestinal uptake process by dietary substrate level is not known and was examined in this study. METHODS: RF uptake was examined using intestinal brush border membrane vesicles (BBMV). RESULTS: Oversupplementation with RF caused significant decrease (P < 0.05; down regulation) in the uptake of 3 mumol/L RF compared with control. This effect was not due to differences in the relative purity of the intestinal BBMV preparations and appeared to be specific for RF. This down-regulation was mediated through a significant (P < 0.01) decrease in the Vmax of the RF uptake process with no significant change in the apparent Michaelis constant (Km). In contrast, RF deficiency caused a significant (P < 0.01) enhancement (up-regulation) in the uptake of 3 mumol/L RF compared with pair-fed control. Again the enhancement was not due to differences in the relative purity of the BBMV preparations and appeared to be specific for RF. This up-regulation was mediated via a significant (P < 0.01) increase in the Vmax of the RF uptake process with no significant change in the apparent Km. CONCLUSIONS: These findings show that the RF intestinal uptake process is regulated by the level of the vitamin in the diet (and/or body stores) and that the regulation is mediated via changes in the number (and/or activity) of the RF uptake carriers with no change in their affinity. PMID- 8224634 TI - Exclusion of APC and MCC as the gene defect in one family with familial juvenile polyposis. AB - BACKGROUND: In familial juvenile polyposis, multiple juvenile polyps occur throughout the colon. The genetic defect has not been characterized. The risk of colon cancer is increased, although the magnitude of the increased risk is controversial. The hypothesis of this study was that the genetic defect is within a tumor suppressor gene, possibly one already known to be inactivated in colorectal neoplasia. METHODS: Linkage analysis using the short tandem repeat polymorphism D5S346 was performed to determine if juvenile polyposis was linked to either APC (adenomatous polyposis coli) or MCC (mutated in colorectal carcinoma) genes within a single large family. RESULTS: A family in which eight subjects have been affected by juvenile polyposis over three generations is described. Six affected subjects had colectomies in childhood, but the two who have so far survived beyond 35 years of age have developed adenocarcinoma of the jejunum. Within this family, linkage analysis excluded linkage of the juvenile polyposis trait to either APC or MCC. CONCLUSIONS: In a family with juvenile polyposis with a clear predisposition to malignancy, including carcinoma of the jejunum, APC and MCC were not the defective genes causing the condition. PMID- 8224635 TI - Effects of fish oil on rectal cell proliferation, mucosal fatty acids, and prostaglandin E2 release in healthy subjects. AB - BACKGROUND: Experimental studies have indicated dietary fish oil as a protective agent in colon carcinogenesis. Prostaglandins have been suggested to be involved in this process. In the present study, the effects of fish oil on rectal cell proliferation (i.e., intermediate biomarker of cancer risk), mucosal membrane fatty acids, and prostaglandin E2 (PGE2) release were investigated in 12 healthy volunteers. METHODS: In addition to a controlled basal diet, the test subjects received either fish oil (4.4 g omega-3 fatty acids/day) or corn oil supplements for two 4-week periods in a double-blind, crossover trial. Rectal cell proliferation was determined by bromodeoxyuridine immunohistochemistry and ornithine decarboxylase activity. After 2-hour incubation with bromodeoxyuridine, PGE2 concentration in the incubation medium was measured by radioimmunoassay. Mucosal membrane fatty acids were analyzed by gas chromatography. RESULTS: Bromodeoxyuridine labeling index (9.2% vs. 10.9%; P < 0.05), ornithine decarboxylase activity (19.7 vs. 36.4 pmol.mg protein-1.h-1; P < 0.005), and PGE2 release from rectal biopsy specimens (435.5 vs. 671.5 pg/mg wet tissue; P < 0.05) were significantly lower during the fish oil than the corn oil period, whereas membrane fatty acids were not statistically different. CONCLUSIONS: The results support the hypothesis that dietary fish oil may protect against colon cancer. PMID- 8224636 TI - Cytokine modulation of intestinal epithelial cell restitution: central role of transforming growth factor beta. AB - BACKGROUND: After various forms of superficial injury, mucosal integrity is re established by rapid migration of epithelial cells across the wound margins in a process termed restitution. The aim of the present study was to assess the role of several regulatory peptides produced within the intestinal mucosa in epithelial restitution. METHODS: The effects of various cytokines and peptide growth factors were studied in an in vitro model of intestinal epithelial restitution. Standard "wounds" were established in confluent monolayers of the intestinal cell line IEC-6, and migration was quantitated in the presence or absence of the physiologically relevant cytokines transforming growth factor (TGF)-alpha, epidermal growth factor (EGF), interleukin (IL)-1 beta, IL-6, tumor necrosis factor (TNF)-alpha, interferon gamma (IFN-gamma), and platelet-derived growth factor (PDGF). RESULTS: Four factors (TGF-alpha, EGF, IL-1 beta, and IFN gamma) enhanced epithelial cell restitution by 2.3-fold to 5.5-fold. In contrast, IL-6, TNF-alpha, PDGF, and an endotoxin lipopolysaccharide had no effect on cell migration. Enhancement of restitution was independent of proliferation. The restitution-promoting cytokines TGF-alpha, EGF, IL-1 beta, and IFN-gamma increase the production of bioactive TGF-beta 1 peptide in wounded IEC-6 cell monolayer. The promotion of IEC-6 restitution by various cytokines could be completely blocked by addition of immunoneutralizing anti-TGF-beta 1. CONCLUSIONS: These findings suggest that various cytokines that are expressed in intestinal mucosa promote epithelial restitution after mucosal injury through increased production of bioactive TGF-beta 1 in epithelial cells. PMID- 8224637 TI - Squamous dysplasia and early esophageal cancer in the Linxian region of China: distinctive endoscopic lesions. AB - BACKGROUND: Linxian, China, has one of the highest rates of esophageal cancer in the world. To design a logical biopsy strategy for large-scale endoscopic surveys in Linxian, the aim of this study was to determine whether squamous dysplasia and early squamous cancer are associated with visible lesions that can be targeted for biopsy. METHODS: Sixty-three Linxian patients with balloon cytological evidence of squamous dysplasia or early cancer of the esophagus had biopsy specimens taken every 4 cm and additional specimens taken from all visually abnormal areas. The appearance of the 398 biopsy sites was described, and abnormal-appearing areas were photographed. The endoscopic descriptions were then compared with the biopsy diagnoses. RESULTS: Twenty-five of 31 (81%) moderately dysplastic or worse specimens (including all nine specimens of invasive cancer) came from visually abnormal sites classified as friability, focal red area, erosion, plaque, or nodule. Fifteen of 16 (94%) patients with moderate dysplasia or worse biopsy diagnoses would have been identified if only these visible target lesions had been sampled. CONCLUSIONS: For surveillance in this high-risk population, random biopsy specimens may be unnecessary; sampling the target lesions described appears sufficient to detect nearly all invasive cancer and most dysplasia. Awareness of these lesion appearances may also aid in earlier detection of squamous cancers of the esophagus in lower-risk populations such as those in Europe and North America. PMID- 8224638 TI - Characterization of muscarinic receptors on guinea pig gallbladder smooth muscle. AB - BACKGROUND: Cholinergic agonists are of major importance for the regulation of gallbladder motility. However, the gallbladder muscarinic receptors have not been localized or characterized directly using radioligands, and it has not been clearly established which subtype of muscarinic receptor mediates contraction. The aim of the present study was to characterize the gallbladder muscarinic receptors. METHODS: Binding studies to guinea pig gallbladder sections were performed using 1-[N-methyl-3H] scopolamine methyl chloride. Carbachol-induced contraction was measured using muscle strips. RESULTS: Binding of 1-[N-methyl-3H] scopolamine methyl chloride was reversible, dependent on time, temperature, and pH. Autoradiography showed binding only over the smooth muscle. Binding and carbachol-induced contractions were inhibited by muscarinic receptor antagonists with the following potencies: atropine > N-methyl-scopolamine > silahexocyclium methylsulfate > AF-DX 384 [(+-)-5,11-dihydro-11-([(2-(2-[(dipropylamino)-methyl] 1- piperidinyl)ethyl)amino]carbonyl)-6H-pyrido (2,3b) (1,4)-benzodiazepine-6-one] > hexahydro-siladifenidol hydrochloride > AF-DX 116 [(+-)-11-([2 [(diethylamino)methyl]-1- piperidinyl]-acetyl)-5,11-dihydro-6H-pyrido (2,3b)(1,4)benzodiazepine-6-one] > pirenzepine. Carbachol inhibited binding to gallbladder sections over the same range of concentrations that caused contractions. The concentration-contraction curves for carbachol were not altered by tetrodotoxin. CONCLUSIONS: Gallbladder smooth muscle cells possess muscarinic receptors of the M3 type. These receptors mediate carbachol-induced contraction. PMID- 8224639 TI - Differential binding of lectins to M cells and enterocytes in the rabbit cecum. AB - BACKGROUND: The afferent limb of the intestinal immune system is represented by the gut-associated lymphoid tissue, in which antigenic material, including complete bacteria, is taken up from the lumen by specialized epithelial cells (M cells). Because the adherence of micro-organisms to epithelial can be mediated by lectin-sugar bindings, the glycoconjugates of the surfaces of M cells and enterocytes were compared. METHODS: A set of 28 lectins and corresponding sugars was used for light and electron microscopy of fixed and unfixed sections. M cells were identified by anti-vimentin antibodies. RESULTS: M cells of the cecal lymphoid patches selectively bound lectins specific for fucose or N acetylgalactosamine. The labeled glycoconjugates were located in the apical membrane and in the membrane of vesicles in the apical cytoplasm. Enterocytes were selectively labeled by galactose-specific lectins. In contrast, the lectin reactivity of M cells and enterocytes did not differ in the jejunal Peyer's patches. CONCLUSIONS: The results suggest that there may be selectivity mediated by glycoconjugates in the uptake of antigenic material by cecal M cells but not by jejunal M cells. PMID- 8224640 TI - Sphincterlike thoracoabdominal high pressure zone after esophagogastrectomy. AB - BACKGROUND: The contribution of the crural diaphragm to the gastroesophageal high pressure zone (HPZ) may be important in prevention of gastroesophageal reflux. The purpose of this study was to investigate the manometric characteristics of the thoracoabdominal junction in patients after surgical removal of the lower esophageal sphincter. METHODS: Ten patients with prior esophagogastrectomy were studied manometrically. RESULTS: Esophageal manometry showed a HPZ and pressure inversion point distal to the anastomosis in 9 of 10 patients. Midrespiratory and end expiratory pressures were 14 +/- 7 and 6 +/- 4 mm Hg above intra-abdominal pressure, respectively. Breath holding caused inhibition of the phasic pressure component. This HPZ relaxed partially in response to deglutition (60% +/- 22%) and contracted in response to increased intra-abdominal pressure induced by either leg lifts or abdominal compression (delta HPZ/delta intra-abdominal pressure = 1.87 +/- 0.64 and 1.96 +/- 0.40, respectively). CONCLUSIONS: This study shows an HPZ at the thoracoabdominal junction after surgical removal of the lower esophageal sphincter. We suggest that this sphincterlike HPZ is due to the crural diaphragm. PMID- 8224641 TI - Distribution and characterization of vasoactive intestinal polypeptide binding in canine lower esophageal sphincter. AB - BACKGROUND: Vasoactive intestinal polypeptide (VIP) may be a nonadrenergic, noncholinergic inhibitory transmitter in the lower esophageal sphincter (LES). There is no biochemical evidence of VIP receptors in the LES. METHODS: Using membranes from canine LES, VIP receptor distribution and characterization were analyzed by radioligand binding and cross-linking experiments. RESULTS: High densities of saturable VIP receptors were found (maximum bound [Bmax], 539.2 fmol/mg in the synaptosome-enriched fraction [P2] and 732.7 fmol/mg in the smooth muscle, plasma membrane-enriched fraction [Mic II]), with high affinity for 125I VIP (dissociation constant [Kd], 1.38 nmol/L in P2 and 1.40 nmol/L in Mic II). Competition binding studies suggested the presence of two binding sites, a high affinity (inhibitor constant [Ki1], 0.064 nmol/L) and a low-affinity (Ki2, 2.68 nmol/L) binding site in P2 membranes, but only one binding site (Ki, 1.18 nmol/L) in Mic II membranes. Guanosine triphosphate-gamma-s pretreatment eliminated high affinity binding in P2 membranes by conversion to binding sites of lower affinity (Ki, 2.82 nmol/L). Studies with a cross-linking agent identified VIP receptors in synaptosomal and smooth muscle plasma membrane fractions; a single polypeptide of approximately 60 kilodaltons was found in each membrane. CONCLUSIONS: Specific VIP receptors exist in both synaptosomal and smooth muscle plasma membrane of canine LES. PMID- 8224642 TI - Lack of discriminant value of dyspepsia subgroups in patients referred for upper endoscopy. AB - BACKGROUND: The subdivision of undiagnosed patients with dyspepsia into symptomatic subgroups (ulcerlike, dysmotilitylike, refluxlike, and nonspecific) may give a clue to the underlying cause; however, the value of this approach in practice is unclear. This study aimed to determine the discriminant value of dyspeptic symptoms and combinations of these symptoms. METHODS: A consecutive sample of 820 outpatients (median age, 62 years; 47% male) completed, before endoscopy, a validated questionnaire that measured 46 gastrointestinal symptoms. RESULTS: Of patients with functional dyspepsia (n = 162), 17% had ulcerlike, 9% dysmotilitylike, and 16% refluxlike dyspepsia alone; 31% fell into two or more symptom subgroups; and 27% had nonspecific symptoms. A similar distribution of the dyspepsia subgroups was observed in patients without functional dyspepsia. Younger age, female gender, frequent upper abdominal pain, no (or sometimes) pain relief with antacid use, and infrequent vomiting were predictive of functional dyspepsia vs. all other diagnoses (at a specificity of 80%, the sensitivity was 60%). The dyspepsia subgroups were poor discriminators in a separate model for functional dyspepsia vs. all other diagnoses (at a specificity of 80%, the sensitivity was only 43%). CONCLUSIONS: The dyspepsia subgroups, as currently defined, appear to have little clinical utility and may be an inappropriate way of classifying dyspepsia. PMID- 8224643 TI - Shape, volume, and content of the deglutitive pharyngeal chamber imaged by ultrafast computerized tomography. AB - BACKGROUND: Conventional radiographic techniques image only the silhouettes of the deglutitive pharyngeal chamber. This study aimed to accurately image the horizontal plane shape and content of the pharynx during swallowing. METHODS: Dynamic computerized tomography images of the pharynx were obtained at the rate of 17 per second during swallowing. Multiple adjacent levels were imaged in eight subjects and a single level was scanned in four subjects during swallows of varied volume. Images were analyzed for area, volume, and the bolus fraction of the deglutitive pharyngeal chamber. RESULTS: The deglutitive chamber enlarged to approximately 24 mL (during tongue loading) compared with a preswallow pharyngeal volume averaging 15 mL. Throughout the 10 mL swallows, the bolus occupied less than 30% of the lumen regardless of axial level. The bolus fraction of the deglutitive chamber increased with swallow volume, as did the dimensions of the upper esophageal sphincter and the bolus velocity through the upper esophageal sphincter. CONCLUSIONS: The deglutitive pharyngeal chamber was typically approximately 15 mL > the bolus volume, implying that an obligatory 15 mL of air was swallowed under these test conditions. Most swallowed air originated as air trapped within the pharynx and larynx as the oropharynx was sealed from above and below. PMID- 8224644 TI - Detection of malabsorption of low doses of carbohydrate: accuracy of various breath H2 criteria. AB - BACKGROUND: Although the accuracy of breath H2 testing to detect malabsorption of small (< 50 g) doses of carbohydrate has never been evaluated, studies suggest that its accuracy is limited by a high prevalence of "H2 nonproducers." The aim of this study was to determine the accuracy of H2 breath testing in the detection of malabsorption of 10 g of carbohydrate. METHODS: In 55 healthy subjects, we determined the ability of breath H2 measurements to distinguish between the ingestion of 10 g of a nonabsorbable carbohydrate (lactulose) and two control meals, a nonabsorbable electrolyte solution or glucose (10 g). RESULTS: The conventional criterion of a 20 ppm increase in breath H2 had 100% specificity but failed to identify lactulose malabsorption in 47% and 24% of subjects at 4 and 8 hours of testing. In contrast, a breath H2 of > 6 ppm at 5 or 6 hours had near perfect specificity and sensitivity. A sum of breath H2 at 5, 6, and 7 hours of > 15 ppm yielded perfect separation between lactulose and control solutions. CONCLUSIONS: True "H2 nonproducers" are extremely rare. Using appropriate criteria, breath H2 testing provides a very accurate means of identifying malabsorption of low doses of carbohydrate. PMID- 8224645 TI - T-cell activation in human intestinal mucosa: the role of superantigens. AB - BACKGROUND: Superantigens are a class of potent T-cell mitogens that activate T cells using specific antigen receptor V regions. Superantigens have been implicated in the pathogenesis of several autoimmune diseases, including inflammatory bowel disease. The present study examines the role of superantigens in the human gastrointestinal immune response. METHODS: Human intestinal epithelial cells and T lymphocytes were obtained from surgical specimens and cultured in the presence or absence of exogenous superantigens. Activation of T cells and V region usage were measured by thymidine incorporation and by cell staining using a panel of monoclonal antibodies. RESULTS: Neither epithelial cells from normal nor diseased intestinal mucosa expressed virally encoded, endogenous superantigens. However, 50% of epithelial cell preparations could present exogenous bacterial superantigens to T cells. In the other 50%, a defect in this function was observed, which did not represent production of suppressive factors or absence of accessory cytokines. Mucosal T lymphocytes proliferated in response to superantigens in vitro, expressing increased transferrin receptor, interleukin-2 receptor, and HLA-DR. CONCLUSIONS: A superantigen-driven mucosal immune response may occur in health and in chronic inflammatory states. The intestinal epithelial cell may mediate this response, through presentation of superantigens to mucosal T lymphocytes. PMID- 8224646 TI - Peptide YY is a physiological regulator of water and electrolyte absorption in the canine small bowel in vivo. AB - BACKGROUND: Peptide YY (PYY), a hormone released following a meal, is one potential mediator of intestinal absorption. Although PYY inhibits 5'-cyclic adenosine monophosphate (cAMP)-stimulated small intestinal secretion in vitro, its effects on fluid and electrolyte transport in vivo are unknown. METHODS: This study examines the effects of physiological doses of PYY in dogs (n = 6) with jejunal and ileal exteriorized, neurovascularly intact intestinal loops (Thiry Vella fistulas). RESULTS: Plasma PYY levels increased after a meal from 155 +/- 15 to 324 +/- 26 pmol/L at 30 minutes and remained elevated for 2 hours. PYY infused intravenously in unfed animals at 25, 50, 100, and 200 pmol.kg-1.h-1, produced a dose-dependent increase in plasma PYY levels. At 100 pmol.kg-1.h-1, PYY plasma concentrations were similar to those of fed animals (317 +/- 39 pmol/L). PYY infusion resulted in a dose-dependent increase in water and electrolyte absorption at all doses in both the jejunum and ileum. Although the relative increase in absorption was similar, the magnitude was greater in the ileum. CONCLUSIONS: Physiological concentrations of PYY produced an increase in small bowel absorption of water and electrolytes in vivo. The postprandial release of PYY may mediate the increase in absorption following a meal. Such a proabsorptive agent may have considerable potential for clinical use in malabsorptive states. PMID- 8224647 TI - Lack of intratumoral heterogeneity in DNA ploidy pattern of hepatocellular carcinoma. AB - BACKGROUND: From biological and clinical perspectives, it is important to clarify tumor heterogeneity. This study was aimed to investigate whether or not intratumoral heterogeneity of DNA ploidy pattern exists in hepatocellular carcinoma (HCC). METHODS: Using fresh materials resected from 31 untreated patients, DNA ploidy was analyzed at different sites of the same HCC by means of flow cytometry. The tumor size ranged from 1.7 to 25.0 cm. RESULTS: There was no case in which euploid and aneuploid HCCs coexisted in the same tumor. The DNA ploidy pattern was euploid in 15 and aneuploid in 16 instances. Of 15 euploid tumors, the areas analyzed were all diploid in 12 and tetraploid in 2 but diploid/tetraploid in 1. Among 16 aneuploid tumors, the DNA indices (DI) at different sites were similar in 9, but apparently, different aneuploid subclones coexisted in 7 cases. The incidence of DI heterogeneity in aneuploid HCCs was similar between small (< 5 cm) and large (> or = 5 cm) tumors; 3 of 7 (42.9%) versus 4 of 9 (44.4%). CONCLUSIONS: It is assumed that euploid and aneuploid HCCs develop in their own ploidy pattern and that the evolution of aneuploid subpopulations from euploid HCC is rare, but new aneuploid subclones can evolve from aneuploid HCC due to increased instability of its karyotype. PMID- 8224648 TI - Evaluation of University of Wisconsin cold-storage solution in warm hypoxic perfusion of rat liver: the addition of fructose reduces injury. AB - BACKGROUND: University of Wisconsin cold-storage solution (UW solution) has markedly improved organ preservation for liver transplantation. However, the efficacy of this solution in preserving hepatocyte viability during warm ischemia is undefined; hence, the effects of UW solution on warm hypoxic injury in the isolated perfused rat liver were examined. METHODS: Livers were perfused using a modified protocol that included a period of hypoxic perfusion with isosmotic Krebs' solution at the end of each experiment. Hepatic injury was evaluated by aspartate aminotransferase (AST) release into the perfusate and the trypan blue perfusion technique. RESULTS: Although UW solution appeared to decrease hepatic injury during hypoxic perfusion, as reflected by low AST release, perfusion with UW solution led to hepatocyte shrinkage and cessation of bile flow even under oxygenated conditions. UW solution did not protect against warm hypoxic injury, as assessed by AST release into the perfusate (182 +/- 15 U/mL, mean +/- SD) or trypan blue staining of the dead hepatocyte nuclei (56% +/- 5%). However, the addition of fructose to UW solution resulted in a significant decrease in AST release (66 +/- 15 U/mL) and parenchymal cell death (39% +/- 7%). CONCLUSIONS: These data suggest that the addition of fructose or other gluconeogenic substrates may complement the overall hepatoprotective effects of UW solution, particularly during periods of warm hypoxia. PMID- 8224649 TI - Administration of N omega-nitro-L-arginine ameliorates portal-systemic shunting in portal-hypertensive rats. AB - BACKGROUND: Nitric oxide, a vasodilator synthesized from L-arginine by vascular endothelial cells, may play a role in the development of portal-systemic collaterals. This study investigated the effect of long-term inhibition of NO secretion on portal systemic shunting. METHODS: Systemic and splanchnic hemodynamics and the degree of portal-systemic shunting were evaluated in partial portal vein-ligated rats after administration of placebo (0.9% saline) or N omega nitro-L-arginine (NNA) (approximately 2 micrograms.kg-1 x min-1) intravenously for 6 days. RESULTS: NNA treatment induced increases in splanchnic arterial resistance (P < 0.001) and portal-collateral resistance (P < 0.05) and a decrease in portal venous inflow (P < 0.05). Portal pressure was not changed (NS). The splenic-systemic shunting was significantly decreased from 81% +/- 5% in the placebo-treated group to 69% +/- 4% in the NNA-treated group (P < 0.05), paralleled by an insignificant reduction in the mesenteric-systemic shunting (64% +/- 7% vs. 50% +/- 6%, NS). The attenuation of portal-systemic shunting by NNA was further shown by an increase in the vascular resistance of portal-systemic collateral venous bed using an in situ portal-systemic collateral perfusion model (1.27 +/- 0.05 vs. 1.07 +/- 0.03 cm H2O.mL-1 x min-1; P < 0.001). CONCLUSIONS: The results show that in portal hypertensive rats, NNA reduces portal-systemic shunting without reducing portal pressure, suggesting that NO plays a role in the collateralization of the portal system. In addition, high flow through the portal collateral bed is probably an important driving force that is independent of portal hypertension for the development of portal-systemic shunting in portal hypertensive rats. PMID- 8224650 TI - Multiple hepatocellular carcinomas at the early stage have different clonality. AB - BACKGROUND: The clonality of hepatocellular carcinoma is still controversial. To evaluate the clonality of this cancer, mostly small multiple hepatocellular carcinomas of 18 patients were analyzed by DNA fingerprinting. METHODS: DNA from the liver tissues was digested with HinfI. After gel electrophoresis, in gel hybridization with a synthetic oligonucleotide probe GTGGTGGTGGTGGTG was performed. The band patterns were compared among the tumors and the nontumor part. In patients positive for hepatitis B surface antigen (HBsAg), the integration pattern of hepatitis B viral DNA in liver tissues was also analyzed. RESULTS: Seven of the 9 HBsAg-positive patients had tumors with different clonalities by both methods. In the remaining 9 HBsAg-negative patients, 4 had different band patterns in their tumors by DNA fingerprinting. CONCLUSIONS: Multiple hepatocellular carcinomas frequently have different clonalities; this finding reiterates the importance of eliminating the underlying cause and the contributing factors of hepatocarcinogenesis. PMID- 8224651 TI - Ultrasonographic diagnosis of acute alcoholic hepatitis 'pseudoparallel channel sign' of intrahepatic artery dilatation. AB - BACKGROUND: In an ultrasound pilot study of acute alcoholic hepatitis (AAH), parallel tubular structures within the liver subsegments were observed. Pulse Doppler flowmetry revealed that these structures were formed by a dilated hepatic arterial branch and an adjacent portal venous branch. This finding was termed the "pseudoparallel channel sign" (PPCS). The aims of this study were to assess the significance of this sign and show the characteristic ultrasound findings of AAH. METHODS: PPCS was specifically searched for on ultrasonography by two physician operators in consecutive patients (77 AAH, 119 other alcoholic liver disease, 49 nonalcoholic liver disease, and 15 healthy patients). RESULTS: PPCS was observed in 90% of patients with AAH and in 23% of patients with other alcoholic liver disease. This sign was not detected in nonalcoholic liver disease or healthy patients. Biopsy specimens were available in 100 patients, 51 of whom were patients with alcoholism. In those 51 patients, PPCS gave a sensitivity of 82%, a specificity of 87%, and an accuracy of 84% in diagnosing AAH. Patients with criteria of AAH had more segments involved with PPCS than patients without. CONCLUSIONS: PPCS may be an important diagnostic finding in AAH. PMID- 8224652 TI - Hepatic and gastric cytoprotective effects of long-term prostaglandin E1 administration in cirrhotic rats. AB - BACKGROUND: Acute administration of prostaglandin E (PGE) may be cytoprotective for hepatocytes in acute hepatitis and for gastric mucosa in cirrhotic rats. We examined the effects of long-term PGE treatment on liver and stomach in cirrhotic rats. METHODS: Cirrhosis was induced by bile duct ligation. Controls had a sham operation. Half the rats received a PGE1 analogue, misoprostol (PGE1) (10 micrograms orally, daily) on days 1-29 postsurgery, and the others received vehicle only. On day 31, all rats underwent ex vivo gastric chamber procedures. Liver chemistry, portal pressures, and hepatic and gastric tissue levels of prostaglandin E2, leukotriene B4, myeloperoxidase, and collagen were determined. RESULTS: PGE1-treated cirrhotic rats had less hepatosplenomegaly, lower serum alanine aminotransferase levels, and portal pressures and higher arterial pressure than vehicle-treated cirrhotic rats. Hepatic and gastric leukotriene B4, myeloperoxidase and collagen levels were significantly lower in the PGE1-treated compared with vehicle-treated cirrhotic rats. Vehicle-treated cirrhotic rats had greater spontaneous and ethanol-induced gastric damage and failed to show a gastric hyperemic response to ethanol, whereas PGE1-pretreated rats did. PGE1 did not significantly affect sham-operated rats. CONCLUSIONS: Long-term PGE1 administration was cytoprotective for both the liver and gastric mucosa in cirrhotic rats. Clinical trials of PGE in human cirrhosis or portal hypertensive gastropathy may be warranted. PMID- 8224653 TI - Ethanol-specific impairment of T-lymphocyte activation is caused by a transitory block in signal-transduction pathways. AB - BACKGROUND: Immune system derangement is characteristic of alcoholic liver cirrhosis. However, in vitro studies have never clarified the alcohol-induced T lymphocyte dysfunction. The aim of this study was to examine any discrete phenotypical and functional abnormalities and possible impairment in transmembrane signal-transduction pathways that, if present on lymphocytes of patients with alcoholic cirrhosis, would also be reproducible after in vitro ethanol exposure of normal T cells. METHODS: Lymphocytes from 25 patients were analyzed for their in vitro proliferative functions, intracellular Ca2+ fluxes, and inositol 1,4,5-triphosphate (IP3) generation. The same procedures were applied to normal T cells exposed in vitro to ethanol. RESULTS: Lymphocytes failed to respond to anti-CD3 and anti-CD2 after in vitro stimulation, with decreased intracellular Ca2+ mobilization and IP3 generation but showed normal proliferative response to phytohemagglutinin. In vitro ethanol incubation of normal T lymphocytes resulted in rearrangement of the membrane CD45 antigen, favoring the expression of high-molecular-weight isoforms, and showed a poor blastogenic response to anti-CD3 and anti-CD2 with a decrease in intracellular Ca2+ mobilization and IP3 production. After a 6-month period of ethanol withdrawal, some patients had normalization of phenotypic and functional alterations. CONCLUSIONS: The T-lymphocyte response to specific polyclonal activators may be severely impaired in alcohol abusers. However, it seems reversible after a period of controlled ethanol withdrawal. PMID- 8224654 TI - Significance of HLA DR4 in type 1 autoimmune hepatitis. AB - BACKGROUND: HLA DR3 and DR4 have been recognized as independent risk factors for autoimmune hepatitis. We compared the clinical features and prognosis of patients with HLA DR4 to those with HLA DR3 and other phenotypes to determine if subclassification by HLA is a valid consideration. METHODS: Forty-four patients with HLA DR4; 41 patients with HLA DR3; and 16 patients with neither allele were studied. Ninety patients were treated with corticosteroids. RESULTS: Patients with HLA DR4 were older (51 +/- 2 years vs. 38 +/- 3 years, P = 0.0001) and more commonly women (89% vs. 68%, P = 0.04) than counterparts with HLA DR3. Additionally, these patients had higher serum immunoglobulin G levels (3300 +/- 216 mg/dL vs. 2732 +/- 192 mg/dL, P = 0.05) and a greater frequency of concurrent immunologic diseases (59% vs. 27%, P = 0.005). Similar differences in clinical presentation distinguished the patients with HLA DR4 from those with other phenotypes. Remission during corticosteroid therapy (85% vs. 63%, P = 0.05) occurred more commonly in the patients with HLA DR4 than in those with HLA DR3 and treatment failure (10% vs. 32%, P = 0.03) occurred less frequently. CONCLUSIONS: Patients with HLA DR4 have a different clinical profile than counterparts with other phenotypes, and they have a better response to corticosteroid therapy than patients with HLA DR3. Subclassification of patients by HLA DR phenotype may have clinical and prognostic value. PMID- 8224655 TI - Propranolol versus sclerotherapy in preventing variceal rebleeding: a randomized controlled trial. AB - BACKGROUND: Sclerotherapy has been widely recommended as initial treatment for prevention of variceal rebleeding. The present study was aimed at comparing the efficacy of endoscopic sclerotherapy and long-term administration of propranolol in the prevention of rebleeding and long-term survival in patients who had bled from varices. METHODS: One hundred sixteen consecutive cirrhotic patients admitted because of variceal bleeding were randomly allocated to either continuous administration of propranolol to reduce the resting heart rate by 25% (58 patients) or weekly intravariceal sclerotherapy sessions using 5% ethanolamide oleate until varices disappeared (58 patients). Results were analyzed on an intention-to-treat basis. RESULTS: Rebleeding occurred in 37 patients of the propranolol group and in 26 patients of the sclerotherapy group (RR = 1.45; 95% CI, 1.03-2.03). The actuarial probability of rebleeding was lower in the sclerotherapy group (P = 0.02). No differences were found in rebleeding index, hospitalization requirements, survival, and causes of death. Complications were significantly more frequent and severe in the sclerotherapy group. CONCLUSIONS: Despite the higher efficacy of sclerotherapy decreasing the probability of rebleeding when compared with propranolol, no beneficial effects were observed on other parameters also reflecting the efficacy of therapy. Moreover, complications of sclerotherapy were more frequent and severe than those of propranolol, which probably shall restrict the use of long-term elective sclerotherapy. PMID- 8224656 TI - Energy metabolism in acute hepatic failure. AB - BACKGROUND: Conflicting data are available concerning energy metabolism in liver disease. Changes should be most pronounced in acute hepatic failure in which loss of 85% of liver cell mass is reported. Metabolic rate could be decreased due to impairment in liver mass but may also be increased as a result of systemic mediator actions. To clarify this issue we studied energy metabolism in acute hepatic failure. METHODS: Energy metabolism was evaluated by indirect calorimetry in 12 patients with acute liver failure and 22 sex-, age-, and body size-matched healthy individuals. In controls and 5 patients, studies were performed in the postabsorptive state; the remaining 7 patients received glucose at a rate of 8 mumol/kg body weight.min to prevent hypoglycemia. RESULTS: Resting energy expenditure was increased in acute liver failure compared with healthy controls (5.1 +/- 0.14 kJ.min-1 x 1.73 m-2 vs. 3.97 +/- 0.08 kJ.min-1 x 1.73 m-2; mean +/- SEM; P < 0.001). Respiratory quotient and oxidation rates for major fuels were not different between the total patient-group and controls. In patients without glucose supply, energy derived from fat was higher and from carbohydrate lower than in healthy controls and patients with glucose supply. CONCLUSIONS: Energy expenditure is increased in acute liver failure. Altered substrate oxidation can be normalized by glucose supply. PMID- 8224657 TI - Antibodies to soluble liver antigen, P450IID6, and mitochondrial complexes in chronic hepatitis. AB - BACKGROUND: Antibodies to soluble liver antigen, P450IID6, and the E2 subunits of mitochondrial dehydrogenase complexes occur in autoimmune liver diseases, but their specificities and implications are uncertain. The aims of the present study were to assess the importance of these autoantibodies in different types of chronic hepatitis. METHODS: Sera from 62 patients with autoimmune hepatitis, 37 patients with cryptogenic hepatitis, and 19 patients with chronic hepatitis C were assessed under code by enzyme immunoassay. RESULTS: Antibodies to soluble liver antigen were found in 7 patients with autoimmune hepatitis (11%) and 5 patients with cryptogenic disease (14%). Patients with antibodies to soluble liver antigen were indistinguishable from seronegative counterparts with autoimmune hepatitis. Seropositive patients with cryptogenic hepatitis had autoimmune features, and they responded to corticosteroid therapy. Five patients (8%) with autoimmune hepatitis were seropositive for antibodies to mitochondrial complexes. Three lacked antimitochondrial antibodies. None of the patients had antibodies to P450IID6, and patients with chronic hepatitis C were seronegative for all markers. CONCLUSIONS: Antibodies to soluble liver antigen do not define a distinct subgroup of patients with autoimmune hepatitis. They may be found in some patients with corticosteroid-responsive cryptogenic hepatitis. Antibodies to E2 subunits and P450IID6 are uncommon in adults with chronic hepatitis. PMID- 8224658 TI - Clinical and virological profiles in patients with multiple hepatitis virus infections. AB - BACKGROUND: The "in vivo" interplay between hepatitis B virus (HBV) and hepatitis C virus (HCV) both in terms of replication activity and cytopathic effect on liver cells is poorly understood. The aim of the study was to investigate their reciprocal influence in patients with HBV and HCV coinfection. METHODS: HBV and HCV genomic sequences in the serum and liver of 55 patients with chronic liver disease who were positive for anti-HCV and for markers of HBV were studied. RESULTS: Twenty-five hepatitis B surface antigen-positive patients, without markers of hepatitis D virus (HDV) infection, showed an inverse relation between seropositivity for HCV RNA and for HBV DNA (P < 0.001). HCV genomic sequences were detected in the liver of all patients positive for HBV DNA but negative for HCV RNA in serum. The biochemical activity and the histological severity of liver disease were lower in HCV RNA-positive/HBV DNA-negative patients, compared with HCV RNA (serum)-negative/HBV DNA-positive cases (P < 0.005). Nine of 10 patients with concurrent HDV infection were negative for serum and liver HCV RNA. None of 20 hepatitis B surface antigen-negative/HCV RNA-positive patients with antibodies to HBV had HBV DNA detectable in serum or liver. CONCLUSIONS: Our findings indicate a reciprocal inverse relation between HBV and HCV replication. Patients positive for antibody to HCV with antibodies to HBV usually have no evidence of HBV DNA persistence in the liver. PMID- 8224659 TI - Intrinsic photoaffinity labeling of native and recombinant rat pancreatic secretin receptors. AB - BACKGROUND: Structural characterization of pancreatic secretin receptors has been limited by difficulties in generating suitable radioligands and obtaining sufficient substrate. The aims of this study were to design, synthesize, and characterize high affinity radiolabeled analogues of secretin suitable for "intrinsic" photoaffinity labeling and to clone, express, and characterize the recombinant rat pancreatic secretin receptor. METHODS: The ability of synthetic analogues to stimulate amylase secretion by pancreatic acini was studied. Receptor complementary DNA (cDNA) was cloned by screening a rat pancreatic library with a probe based on the sequence of a neural cell secretin-binding protein. Competition binding and affinity labeling were performed with membranes prepared from rat pancreas and transfected cells. RESULTS: Two probes were fully efficacious secretagogues, which bound in a specific, high-affinity, rapid, and temperature-dependent manner. Only ([125I]Tyr10, pNO2-Phe22) rat secretin 27 covalently labeled a 50,000-62,000-molecular weight pancreatic membrane protein, with labeling inhibited in a concentration-dependent manner by secretin but not vasoactive intestinal polypeptide. Hybridization screening yielded a full-length cDNA identical to the neural clone. Photoaffinity labeling of this recombinant receptor identified a 57,000-62,000-molecular weight protein with specificity similar to that of native pancreas. Both native and recombinant receptors migrated at a molecular weight of 42,000 after endoglycosidase F deglycosylation. CONCLUSIONS: This study provides evidence for the molecular identity of the pancreatic secretin receptor and presents a novel probe important in structural characterization of its agonist-binding domain. PMID- 8224660 TI - Angiostrongyliasis in a European patient: a rare cause of gangrenous ischemic enterocolitis. AB - Angiostrongylus costaricensis causes a clinicopathologic disease first observed in Costa Rica, mainly in children characterized by highly symptomatic eosinophilic gastroenteritis involving the terminal ileum, cecum, appendix, and ascending colon. A case of angiostrongyliasis in an adult Spaniard infected during a brief stay in Nicaragua is reported. We examined the macro-microscopic features of a right ileocolic resection of a 52-year-old patient. In the surgical specimen, multiple confluent ulcers, some perforated, were observed particularly in the terminal ileum. The intestinal wall was thickened, and the arterial branches presented necrotizing arteritis with thrombosis and a heavy infiltrate of eosinophils around the vessels. In addition, some granulomas were observed. Adult male and female forms of A. costaricensis were found in the lumen of many arterial branches and in the tissue of the omentum, surrounded by dense eosinophilic infiltrate. A. costaricensis causes an intense eosinophilic, necrotizing arteritis associated with thrombosis leading to severe ischemic lesions. This is the first case to be diagnosed in Europe. Incubation time was able to be established within a period of at least 14 days. PMID- 8224661 TI - Nasopharyngeal angiofibroma in patients with familial adenomatous polyposis. AB - Four patients with nasopharyngeal angiofibroma and familial adenomatous polyposis are reported here. Nasopharyngeal angiofibroma was 25 times more frequent in our patient population with familial adenomatous polyposis than in an age-matched hospital population. The association of these two rare conditions suggests that nasopharyngeal angiofibroma is an extracolonic manifestation of adenomatous polyposis. In addition, somatic mutation of the adenomatous polyposis coli gene, which causes adenomatous polyposis when mutated in the germline, could play a role in the pathogenesis of sporadic nasopharyngeal angiofibroma. PMID- 8224662 TI - Arteriovenous malformation at pancreatobiliary region causing hemobilia after cholecystectomy. AB - Pancreatic arteriovenous malformation is a rare condition that may cause gastrointestinal bleeding. A 66-year-old man with large arteriovenous malformation at the pancreatobiliary region is described. The patient had recurrent episodes of hemobilia after cholecystectomy performed for the treatment of cholelithiasis. Enlargement of the arteriovenous malformation was documented by angiography performed before and after the cholecystectomy. Bleeding from the biliary tract was successfully controlled by transarterial embolization. Cholecystectomy may have caused a hemodynamic change at the pancreatobiliary region, leading to the enlargement of the lesion and hemobilia. PMID- 8224663 TI - Chronic constipation in children. AB - The evaluation of chronic constipation with or without encopresis must begin with a careful history. The intervals between bowel movements and the size and consistency of stools deposited into the toilet should be noted. Encopresis may be manifested as dirtying the underwear. The physical examination should include a rectal and neurological examination. No specific organic cause can be found in the majority of children. One or several anorectal physiological abnormalities have been found by us and others in 95% of children with idiopathic constipation. These abnormalities include impaired rectal and sigmoid sensation and decreased rectal contractility during rectal distention. The external anal sphincter and pelvic floor muscles may be abnormally contracted during straining for defecation, and the child may be unable to defecate a rectal balloon. Most patients will benefit from a program designed to clear stools, to prevent further impaction, and promote regular bowel habits. Fifty percent of patients will be cured after 1 year and 65%-70% after 2 years. PMID- 8224664 TI - The crural diaphragm, an external lower esophageal sphincter: a definitive study. PMID- 8224665 TI - The third dimension. PMID- 8224666 TI - Potential role of superantigens in gastrointestinal disease. PMID- 8224667 TI - How many types of autoimmune hepatitis are there? PMID- 8224668 TI - Vegetable protein diet and hepatic encephalopathy. PMID- 8224669 TI - Colorectal carcinogenesis: searching for the lost genes. PMID- 8224670 TI - Ultrasonic sizing of antrum as an indicator of gastric emptying. PMID- 8224671 TI - Gastric ulcers--when is enough enough? PMID- 8224672 TI - Erythromycin and gallbladder motor function. PMID- 8224673 TI - Fundic gland polyps: a rare, innocuous, and reversible disturbance. PMID- 8224674 TI - Differentiation of endothelin-1 and endothelin-3 concentrations. PMID- 8224675 TI - Medically induced achlorhydria and gastric cancer risk. PMID- 8224676 TI - Effects of intraduodenally applied bile salts on pancreatic secretion. PMID- 8224677 TI - Nodular transformation of the liver: a reply. PMID- 8224678 TI - Microscopic colitis cases revisited. PMID- 8224679 TI - Adding a sclerosant to endoscopic epinephrine injection in actively bleeding ulcers: a randomized trial. AB - We compared the efficacy of epinephrine injection and epinephrine injection followed by sodium tetradecyl sulfate in controlling active ulcer bleeding. Out of 2814 patients who underwent endoscopy for gastrointestinal bleeding, 200 patients with actively bleeding ulcers seen at the time of endoscopy were randomized to receive epinephrine injection alone (99 patients) or epinephrine injection followed by 3% sodium tetradecyl sulfate (101 patients). After the procedure the patients were transferred to the surgical gastroenterology ward and were treated by surgeons who were unaware of the mode of treatment. The patients underwent routine endoscopy 24 hours later, and epinephrine injection was repeated if active bleeding was seen again. Emergency surgery was performed for the following: (1) arterial spurting not controlled endoscopically, (2) failure of the blood pressure or pulse to stabilize after 4 units of blood, (3) total transfusion of more than 8 units of blood, or (4) rebleeding as defined by hematemesis with pulse greater than 100 beats/min or blood pressure less than 100 mm Hg after stabilization. The two groups were comparable in age, sex, site of ulcer, and severity of bleeding. Initial hemostasis was obtained at the time of endoscopy in 94% of the epinephrine group and 97% of the epinephrine plus sodium tetradecyl sulfate group. No difference in outcome was seen in the two groups as measured by emergency surgery requirement, blood transfusion, hospital stay, and hospital mortality. Endoscopic epinephrine injection is effective in controlling active ulcer bleeding. The additional injection of sodium tetradecyl sulfate confers no additional advantage. PMID- 8224680 TI - Cutting the defiant lower esophageal ring. AB - A symptomatic lower esophageal ring generally responds to standard bougienage therapy. However, a subset of patients with a lower esophageal ring defy dilation therapy. We have evaluated the efficacy of electrocautery incision of "defiant" lower esophageal rings, the data of which forms the basis of this report. Seven of 75 patients with a lower esophageal ring failed to respond to conventional esophageal dilation. All seven patients underwent electrocautery incision of a "defiant" lower esophageal ring with alleviation of dysphagia. One patient had recurrence of dysphagia that occurred 6 months after initial electrocautery incision. No major complication occurred, although one patient had transient chest pain. Subsequently, all patients have remained without symptoms at a mean follow-up of 36 months. Electrocautery incision of "defiant" lower esophageal ring is efficacious, safe, and results in long-term relief of dysphagia. PMID- 8224681 TI - Evaluation of complications during and after conscious sedation for endoscopy using pulse oximetry. AB - All events prolonging an endoscopic procedure or recovery, or requiring a medication or an intervention, were analyzed from a consecutive sample of 508 patients receiving conscious sedation. Although 102 events were identified (20%), 33 of these (7%) were major. These included four episodes of apnea and four patients with a prompt and sustained fall in oxygenation during the procedure; 19 additional patients had a decrease to less than 89% in oxygen saturation in the 30 minutes after the completion of the procedure. The patients with observed events had significantly more major illnesses, a higher fraction older than 70 years, and a higher fraction of endoscopic retrograde cholangiopancreatography than those without events (p < 0.05) but had a similar dose of sedative medications, mean age, and fraction of colonoscopies. PMID- 8224682 TI - Techniques and complications of esophageal foreign body extraction in children and adults. AB - To assess the success rates and complications of esophageal foreign body extraction in adult and pediatric patients, the charts of 76 adults and 116 children with endoscopically or radiographically documented esophageal foreign body impaction were retrospectively reviewed. Success rates for rigid esophagoscopy (100%) and flexible endoscopy (96.2%) were not significantly different (p > 0.05). Overall, rigid esophagoscopy had a higher complication rate than flexible endoscopy (10% versus 5.1%, p > 0.05), but this trend did not reach statistical significance. The Foley catheter technique was used predominantly in children with proximally located blunt objects. Our study shows that flexible and rigid esophagoscopy are both safe and effective methods of removing esophageal foreign bodies, but rigid esophagoscopy carries a higher complication rate and therefore should be reserved for patients in whom flexible endoscopy is unsuccessful. The Foley catheter technique is suited only for proximally located blunt objects, and its routine use is not recommended. PMID- 8224683 TI - Prospective, randomized trial comparing sodium phosphate solution with polyethylene glycol-electrolyte lavage for colonoscopy preparation. AB - We performed a prospective randomized trial in a predominantly outpatient colonoscopy population to see how preparation with oral sodium phosphate solution compares with polyethylene glycol-electrolyte lavage in terms of the quality of colon cleansing, ease of preparation, and gastrointestinal intolerance. Before colonoscopy, a nurse administered a questionnaire to the patient to assess how well the preparation was tolerated (scale from 1 to 5:1 = easy, to 5 = unable to finish) and about the presence of four symptoms: abdominal pain, nausea, vomiting, and dizziness. The quality of colon cleansing was graded by the attending gastroenterologist, who was unaware of how the patient was prepared or tolerated the preparation (1 = excellent, 2 = good, 3 = fair, 4 = poor). The overall quality of bowel preparation with polyethylene glycol lavage was slightly better than with sodium phosphate (mean score, 1.93 vs 2.07); however, the difference was not statistically different. No statistical difference was seen in the frequency of patients with poor preparations (14.2% for sodium phosphate, 9.6% for polyethylene glycol lavage). Patients found preparation with sodium phosphate to be somewhat easier than polyethylene glycol lavage (mean score, 2.07 vs 2.41; p = 0.05). No difference was seen in the incidence of abdominal pain, nausea, or vomiting. Dizziness was more common with sodium phosphate but was mild and not believed to be clinically important. We conclude that the quality of colon cleansing is similar with polyethylene glycol lavage and oral sodium phosphate solution, with satisfactory preparation seen in 85% to 90% of patients. Patients found preparation with sodium phosphate to be slightly easier to tolerate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224684 TI - Endoscopic ultrasonographic evaluation of the rectum in cirrhotic portal hypertension. AB - Rectal endoscopic ultrasonography was performed using an ultrasound fiberscope in 20 patients with cirrhotic portal hypertension (6 alcoholic patients, 4 patients with hepatitis B surface antigen positive, and 10 cryptogenic patients) and in 10 patients with irritable bowel syndrome as controls. Rectal varices were diagnosed endoscopically when either tortuous or saccular distended veins were seen beneath the mucosa. At rectal endoscopic ultrasonography rectal varices were seen as rounded or oval echo-free structures in the submucosa. Rectal endoscopic ultrasonography also showed perirectal veins outside the rectal wall. Rectal varices were detected by endoscopy in 9 patients and by rectal endoscopic ultrasonography in 17 patients. Rectal endoscopic ultrasonography also detected submucosal veins in 3 of 10 controls. The number and size of submucosal veins seen on rectal endoscopic ultrasonography in patients with portal hypertension were greater than in controls (p < 0.01 for both number and size). The size of perirectal veins was greater in patients than in controls (p < 0.05), although their number was no different (p = NS). A perforating vein communicating between a submucosal and perirectal vein was seen in only one patient. Rectal wall thickness was not different in patients and controls (p = NS). Rectal endoscopic ultrasonography was superior to endoscopy in detecting the presence (85% versus 45%, p < 0.01), and number (p < 0.01) of rectal varices. Our study suggests that rectal endoscopic ultrasonography is useful in detecting changes in rectal and perirectal vasculature in patients with cirrhotic portal hypertension. PMID- 8224685 TI - Argon laser therapy for hemorrhagic radiation proctitis: long-term results. AB - In chronic radiation proctitis bleeding occurs from mucosal friability and neovascular telangiectasias. Fourteen patients with bleeding from chronic radiation proctitis underwent endoscopic argon laser therapy at 4 to 8 W. The goal of treatment was obliteration of all telangiectasias. The average follow-up was 35 months. Of the 51 procedures, 48 (94%) were performed on outpatients with enema preparation and little or no sedation. A median of three procedures was performed per patient, with two sessions required for initial control of bleeding. Ten patients (71%) required maintenance therapy for recurrent bleeding from telangiectasias that developed after initial therapy. The mean interval between maintenance sessions was 7 months. No immediate or late complications occurred. PMID- 8224686 TI - Drainage of the gallbladder in patients with acute acalculous cholecystitis by transpapillary endoscopic cholecystotomy. AB - The mortality associated with acute acalculous cholecystitis approaches 50%. Removal or decompression of the gallbladder in these patients may prevent gallbladder rupture and may be lifesaving. This is usually accomplished by cholecystectomy, cholecystotomy, or percutaneous gallbladder drainage. We describe a novel transpapillary endoscopic approach to gallbladder drainage in patients at high surgical risk. A total of seven high surgical risk patients were treated with transpapillary endoscopic cholecystotomy. Cannulation of the cystic duct was accomplished by using standard hourglass-tipped catheters in two patients. A new "selector" catheter was developed for selective cannulation of the cystic duct and used in the other five patients. Five of the seven patients showed evidence of clinical, radiographic, and laboratory improvement after treatment. We conclude that transpapillary endoscopic cholecystotomy may be an effective treatment alternative for high surgical risk patients with acalculous cholecystitis. PMID- 8224687 TI - Effect of prophylactic main pancreatic duct stenting on the incidence of biliary endoscopic sphincterotomy-induced pancreatitis in high-risk patients. AB - Pancreatitis is a common complication of endoscopic sphincterotomy. Cautery induced papillary edema has been implicated as a possible cause. The objective of this study was to determine whether prophylactic stenting of the main pancreatic duct after endoscopic sphincterotomy in high-risk patients would reduce the incidence of pancreatitis. High-risk patients were defined as those with sphincter of Oddi dysfunction, small common bile duct diameter (< 10 mm), or those requiring pre-cut sphincterotomy. Patients were studied in a prospective fashion from October 1990 to April 1992 and were randomized to receive either a main pancreatic duct stent or no stent after biliary sphincterotomy. The stents were generally removed 10 to 14 days after placement. Fifty patients were randomized to the no-stent group and 48 patients were randomized to the stent group, but in five patients stent placement was unsuccessful. Pancreatitis occurred in 18% of patients in the no-stent group compared with 14% of patients in the stent group. Most cases of pancreatitis were mild, occurring in 10% of patients in the no-stent group and 12% of patients in the stent group. Moderate to severe pancreatitis occurred with an increased frequency in patients in the no stent group (8%) compared with that in patients in the stent group (2%). Mean number of hospital days required to treat pancreatitis was 9.5 days in the no stent group compared with 2.8 days in the stent group; however, none of these differences reached statistical significance. Small common bile duct diameter (< 6 mm) was found to be an independent risk factor for pancreatitis after endoscopic sphincterotomy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224688 TI - Polyethylene pancreatic duct stent-induced changes in the normal dog pancreas. AB - This study was undertaken to evaluate the frequency, severity, and reversibility of stent-induced pancreatic ductal and parenchymal changes in the normal dog pancreas. Six adult mongrel dogs underwent duodenotomy and placement of 5F polyethylene pancreatic duct stents into the main pancreatic duct. After 8 weeks of stenting, the animals were randomly assigned to one of three groups: group I (n = 2), sacrifice; group II (n = 2), stent removal followed by sacrifice after an 8-week recovery period; group III (n = 2), the stent was exchanged and kept in place for an additional 8 weeks. In group III at 16 weeks the stent was removed, and the animals were allowed to recover for 8 weeks before sacrifice. Pancreatograms were obtained at each operation and were normal before stent placement. Gross and histologic evaluation was performed at the time of sacrifice. All stented animals developed radiographic, gross, and histologic abnormalities. Pancreatograms showed duct dilation in the stented region associated with a short stenosis at the mid to upstream segment of the stent. Group III animals had more advanced radiographic changes than group I and II animals. The radiographic findings were associated with gross evidence of fibrosis, which increased proportionately with the length of the stenting period. Group III animals developed moderate to severe pancreas atrophy. Histologic changes of obstructive pancreatitis were present in most experimental dogs. The results of this study suggest that pancreatic stenting may cause permanent damage in the normal dog pancreas. The cause of the damage appears to be related to stent occlusion, perhaps exacerbated by local stent-induced trauma. PMID- 8224689 TI - Endoscopic therapy of pancreatitis in the pediatric population. AB - Six children with chronic relapsing pancreatitis or acute post-traumatic pancreatic pseudocyst underwent therapeutic pancreatography at the time of ERCP. Procedures included pancreaticobiliary sphincterotomy, pancreatic duct stone removal, and endoprosthesis placement. Four of five patients with chronic pancreatitis had significant symptomatic improvement, whereas the patient with pseudocyst had rapid resolution of symptoms and cyst. Complications were limited to mild pancreatitis in a subset of patients. With the capability of obviating or delaying the need for palliative pancreatic surgery, therapeutic pancreatography deserves additional evaluation in the pediatric patient. PMID- 8224690 TI - Bacterial adherence to hydrophilic polymer-coated polyurethane stents. AB - Biliary stent blockage by adherence of bacteria and formation of biofilm is a major problem in endoscopic stenting procedures. We have investigated bacterial adherence to hydrophilic polymer (Hydromer)-coated polyurethane stents in stationary and perfusion experiments. Adherence of gram negative and gram positive organisms under stationary conditions was similar between Hydromer coated and non-coated control polyurethane stents; however, a marked 1 to 3 log reduction in adherence to Hydromer-coated stents occurred when perfusion experiments in phosphate buffer solution or bile were performed. The results suggest that Hydromer-coated polyurethane stents could be useful in preventing complications caused by biliary stent blockage. PMID- 8224691 TI - Serial cholangiographic appearances in recurrent pyogenic cholangitis. AB - From December 1982 to December 1991, cholangiograms were obtained in 227 patients with recurrent pyogenic cholangitis. Cholangiographic abnormalities included biliary dilation, calculi, sludge, excessive branching, and arrowhead formation of intrahepatic ducts and biliary strictures. In 21 patients, previous evidence of biliary ascariasis was seen. Repeat cholangiograms were performed in 55 patients in a follow-up period of 18.0 +/- 1 months. Of these patients, 12 treated conservatively continued to get recurrent cholangitis and revealed worsening abnormalities on repeat cholangiograms. Another 25 patients had successful endoscopic sphincterotomy and extraction of biliary calculi. These patients remained free of symptoms on follow-up, with significant resolution of abnormalities on repeat cholangiograms. The remaining 18 patients with failed surgical or endoscopic interventions continued to get recurrent episodes of cholangitis and worsening of abnormalities on repeat cholangiograms. This retrospective study indicates that the natural course of recurrent pyogenic cholangitis is a progressive, destructive cholangiopathy. Ascaris lumbricoides invasion of the biliary tree is an initiating event in a sub-group of patients. PMID- 8224692 TI - Worm extraction and biliary drainage in hepatobiliary and pancreatic ascariasis. AB - From December 1989 to March 1992, 50 (32%) of 156 patients with hepatobiliary and pancreatic ascariasis underwent various endoscopic interventional procedures. Endoscopic intervention was performed when patients did not respond to energetic symptomatic treatment within the first few days of hospitalization (n = 45) or when worms had not moved out of the ducts into the duodenum at 3 weeks (n = 5). Worm extraction was successful in all 18 patients from the ampullary orifice and in 34 (89.5%) of 38 patients from the bile or pancreatic duct. In five patients with pyogenic cholangitis, endoscopic nasobiliary drainage was performed to decompress the bile ducts. After worm extraction/nasobiliary drainage, 41 (91%) of the 45 patients with biliary disease (n = 42) or acute pancreatitis (n = 3) had rapid relief of symptoms. Three patients developed complications related to endoscopy, including cholangitis (n = 2) and hypotension (n = 1). PMID- 8224693 TI - A transparent endoscopic elastic band ligating device. PMID- 8224694 TI - Improved biliary stenting using a balloon catheter and the combined technique for difficult stenoses. PMID- 8224695 TI - Endoscopic needle localization of pancreatic pseudocysts before transmural drainage. PMID- 8224696 TI - A method for the endoscopic retrieval of trichobezoars. PMID- 8224697 TI - Mirror-image artifact with endoscopic ultrasonography and reappraisal of the fluid-air interface. PMID- 8224698 TI - Probable ischemic proctosigmoiditis presenting 8 weeks post-partum. PMID- 8224699 TI - Endoscopic fragmentation of gastric phytobezoar by electrohydraulic lithotripsy. PMID- 8224700 TI - Pancreatic stones as a cause of bile duct and ampullary obstruction: endoscopic treatment approaches. PMID- 8224701 TI - Endoscopic management of common bile duct stones resulting from metallic surgical clips (cat's eye calculi). PMID- 8224702 TI - Primary linitis plastica of the colon. PMID- 8224703 TI - Inflammatory polyps as a manifestation of intestinal graft versus host disease. PMID- 8224704 TI - Why should gastroenterologists know about outcomes research? PMID- 8224705 TI - Endoscopy and the evolution of the GI specialties: ASGE Presidential Address- 1993. PMID- 8224706 TI - Physicians, patients, and health care policy. PMID- 8224707 TI - The ASGE guidelines and health care resources. PMID- 8224708 TI - Enhanced detection of early gastric cancer. PMID- 8224709 TI - Is early endoscopy after percutaneous endoscopic gastrostomy placement dangerous? PMID- 8224710 TI - Colonoscopic diagnosis of mucocele of the appendix. PMID- 8224711 TI - Gastric carcinoma containing a surgical clip: endosonographic appearance. PMID- 8224712 TI - Laser endosurgery on biliary wire mesh stents. PMID- 8224713 TI - Use of a polypectomy snare to salvage a faulted stent placement. PMID- 8224714 TI - Esophageal casts: unusual finding on removal of a nasogastric tube. PMID- 8224715 TI - Colonoscopy and endoscopic therapy of hemorrhage from viral colitis. PMID- 8224716 TI - Helicobacter pylori-associated gastric ulcer mimicking a gastric malignancy. PMID- 8224718 TI - Improved photographic documentation. PMID- 8224717 TI - Pneumopericardium without subcutaneous emphysema, pneumomediastinum, or pneumothorax after laparoscopy. PMID- 8224719 TI - [External fetal version from breech to vertex presentation. Evaluation of 1,000 cases]. AB - Results of 1000 external versions of foetuses from breech to vertex presentation under tocolysis were evaluated. The rate of success of the versions was 52%. The success rate improved with gestational age, with increasing parity, with increasing age of the mother and her increasing body weight. The location of the placenta did not play a decisive role. The success rate of the version was remarkably good, even, in cases with small-for-gestational age foetuses and large for-gestational age foetuses as well as in cases with a previous Caesarean section. The frequency of typical complications and the resulting Caesarean rate of 2.3% are within the acceptable limit. Up to now, from a total of more than 1,500 versions performed at the Department of Obstetrics at the Women's Hospital, Berlin-Neukolln, no death of either mother or child has occurred, which could be related to this procedure. Of course, the prerequisite for a safe version is strict adherence to the essential guidelines. The fact, that versions are not practised in all obstetrical departments, is partly due to the embarrassing lack of expert knowledge on the part of some practitioners and clinicians. In numerous cases, a moderately difficult laparotomy--which Caesarean section is in principle -with all the inevitable risks and dangers, could have been prevented, if a version had been performed. PMID- 8224720 TI - [Doppler ultrasound diagnosis in post-term pregnancy]. AB - The capability of Doppler flow velocimetry to predict intrauterine growth retardation is well known. The increased morbidity and mortality rate of postterm newborns is also well known. The aim of our study was to examine if Doppler flow velocimetry is able to indicate foetal jeopardy in the postterm period. Flow velocimetry of the foetal descending aorta, the umbilical artery, the uterine arteries and in 59 cases also the foetal middle cerebral artery was obtained from 167 pregnancies after 40 completed weeks of gestation. We found significant changes of normal values in prolonged pregnancy compared to third trimester normal values, examining the mean velocity of the foetal descending aorta and the S/D-ratio of the umbilical artery. No clinically significant changes were found examining the S/D-ratio of the uterine arteries and the pulsatility index of the foetal middle cerebral artery. Daily examinations of the foetal descending aorta were carried out in 23 and of the umbilical artery in 19 cases during the last four days before delivery, and in 11 cases of the foetal middle cerebral artery during the last three days before delivery. We did not find significant changes in the medians of the mean velocity of the foetal aorta, of the S/D-ratio of the umbilical artery and of the pulsatility index of the foetal middle cerebral artery. Measurement of sensitivity and positive predictive value of the four arteries examined showed, that Doppler ultrasound could not predict small for date infants or Caesarean section because of foetal distress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224721 TI - [Cesarean section of the second twin. Is this unusual mode of delivery justifiable?]. AB - Within a ten-year period 1983-1992 eleven Caesarean sections were performed on the second twin after the first twin had been born vaginally. The reasons leading to this rather unusual mode of delivery are described here. Predominantly, multiple risk factors were present. One first twin and one second twin died in the early post natal period. In a comparative study of the literature, it is shown, that this, disputed mode of delivery has increased during the last twenty years and that, under special circumstances, it is a justifiable method of management of gemini deliveries. PMID- 8224722 TI - [Effect of pregnancy on the incidence and course of malignant diseases]. AB - Fortunately, coincidence of pregnancy associated with malignant neoplasm is rare. As reported in the literature, incidence is 1.5-10: 10,000 pregnancies. The study presented reports data on the incidence and outcome of 16-35 years old female patients suffering from malignant neoplasms. Patients, who were treated in Freiburg 1980-1989, were considered for evaluation. We analysed the impact of pregnancy on the outcome of these patients by stratifying patients for the time of diagnosis (before, during, or after a pregnancy). 247 patients were included. 118 patients developed a neoplasia after a successful pregnancy. In 24 patients, neoplasia was diagnosed during pregnancy, and 28 patients became pregnant after diagnosis and therapy for a malignancy. Further 77 patients without pregnancy, but in whom neoplasia diagnosed at the age of 16-35 years were included. Cancer of the cervix uteri, breast cancer, ovarian cancer, and malignant lymphomas were the most frequent neoplasias diagnosed in young women. In an analysis stratified for stage of disease, we found no significant difference between 3- and 5-years survival of patients with pregnancies before, during, or after diagnosis and treatment of neoplasia. Due to the inhomogeneity of the subgroups analysed, the question, whether pregnancy has any impact on the outcome of neoplasm could not be conclusively answered. The necessity for the establishment of national and international registries collecting sufficient data about incidence and outcome of patients with pregnancies associated with malignant neoplasms is emphasised. PMID- 8224723 TI - [Marshall-Marchetti-Krantz operation or fasciaplasty in therapy of recurrent urinary incontinence in women]. AB - In this study, a follow-up examination was carried out, comprising 59 female patients who were operated between the years 1985 and 1991 because of recurrent urinary incontinence. We distinguished two different operating methods: 34 women were treated following the Marshall-Marchetti-Krantz operation technique, whereas a fasciaplasty-suspension was employed om 25 patients. At the urodynamic follow up examination, the patients had experienced an average history of 2 years' recurrent urinary incontinence. The patients' subjective observation of anamnestic urinary incontinence was objectified by a clinical cough test, cystotonometry and sonography of the small pelvis. Making extensive use of the urodynamic examination possibilities showed a 79.4% improvement following the Marshall-Marchetti-Krantz operation in comparison to a 52% improvement after the fasciaplasty operation. The operations success is assessed by evaluation of the depression quotient. Here the Marshall-Marchetti-Krantz operation resulted in a significant improvement. After the fasciaplasty operation, however, this was not found to be the case. PMID- 8224724 TI - [Homologous intrauterine insemination as a therapeutic possibility in andrologically-induced sterility]. AB - In a two-years period, we treated 58 infertile couples for a total of 213 cycles with artificial homologous insemination (AIH). We considered a indication for AIH low sperm quality or a pathological postcoital test (PCT). Couples with idiopathic infertility did not receive AIH. If possible, AIH was performed by intrauterine insemination (IUI), following concentration of the motile sperms by "swim-up" method. Out of 213 cycles "swim-up", preparation could not be done in 46 cases (21.6%) because the initial sperm quality was below technical limits (sperm count below 1 Mio/ml). Out of 58 treated couples 4 became pregnant during AIH therapy (7%) among, them 2 couples conceiving twice after spontaneous early abortion. The total pregnancy rate per cycle was 2.8%. In the cycles after "swim up", the monthly pregnancy rate through IUI was 3.6%. In the presence of andrological factors was the pregnancy rate only 0.9% compared to 6.8% in cases of pathological PCT with normal sperm quality. Thus, it can be concluded, that IUI is not successful on the basis of low sperm quality. Only in cases of unfavourable cervical factor expressed by a pathological PCT accompanied with a normal spermiogramm, moderate success can be expected. PMID- 8224725 TI - [Prolactinoma after craniocerebral trauma]. AB - A female patient, now twenty-four years old, suffered from a severe cerebral contusion in 1983 as a result of a car accident. At the time of the accident, she had a regular menstrual cycle. Since 1985, she has contracted oligomenorrhoea and since 1988 secondary amenorrhoea. In 1988, we diagnosed by CT-scan and NMR imaging an excessive hyperprolactinaemia caused by a pituitary adenoma. Neither surgical intervention, nor conservative medication could reduce the prolactin level to normal. We suspect, that there is an association between the cranial injury and the appearance of the pituitary tumour. PMID- 8224726 TI - [Transposition of the external genitalia]. AB - The syndrome of complete peniscrotal transposition is rare. Combination with malformations of other organs is frequent. Sometimes, life is not possible with these malformations. The demonstrated case gives an overview of: Definition, Epidemiology, Etiology and the Diagnostic and therapeutic procedure. PMID- 8224727 TI - [Toxoplasmosis in pregnancy. Comments on the contributions by T. Beringer and T. Krause, W. Straube, S. Wiersbitzky, V. Hitz, A. Kewitsch]. PMID- 8224728 TI - The effect of caffeine on the contractile response of the rabbit urinary bladder to field stimulation. AB - 1. Smooth muscle contraction is mediated by an increase in the concentration of cytoplasmic free calcium. 2. Low concentrations of caffeine can induce transient contraction of smooth muscle through the release of intracellular calcium from intracellular storage sites. 3. The current study determined the effect of caffeine (0.4-100 microM) on the response of the in vitro whole bladder preparation to field stimulation (FS) at 0.6, 1.8, and 5.4 mM extracellular calcium. 4. The following parameters were determined: basal pressure (BP), peak pressure response (PP), maximal rate of pressure generation (PR), maximal rate of bladder emptying (RE), and percent of volume emptied (VE). 5. The results demonstrated that in the absence of caffeine, BP, PP, and PR were increased progressively as a function of both frequency of stimulation and the concentration of calcium. 6. In addition, caffeine significantly increased the rate of pressure generation at 0.6 and 1.8 mm calcium. PMID- 8224729 TI - Antagonism of antidiuretic hormone in domestic pigs (Sus scrofa). AB - 1. Morphological and physiological aspects of renal function are shared by humans and swine. SK&F 101926 is a potent antagonist of vasopressin binding to V2 receptors and vasopressin stimulation of adenylate cyclase activity in renomedullary membranes from both species. 2. Unexpectedly, SK&F 101926 proved to be an antidiuretic agonist in humans. Hence, we evaluated SK&F 101926 for antidiuretic agonist and antagonist activities in conscious domestic pigs. 3. During water diuresis (Uosm < 230 mOsm/kg H2O), administration of SK&F 101926 (100 micrograms/kg, i.v.) produced a maximal Uosm of 192 +/- 18 mOsm/kg H2O, a concentration not significantly different from that in vehicle-treated pigs. 4. In hydropenia, SK&F 101926 produced a modest decrease in Uosm, from 945 to 629 mOsm/kg H2O (P < 0.05). 5. In in vitro studies subsequently performed using renomedullary tissue from the same pigs, SK&F 101926 displayed high affinity for V2 receptors (Kbind = 11.8 nM) and high potency to inhibit vasopressin stimulation of adenylate cyclase (Ki = 3.9 nM). 6. No activity of SK&F 101926 to stimulate adenylate cyclase activity was detected. 7. We conclude that, in spite of its activity in in vitro assays, SK&F 101926 is a weak antidiuretic antagonist in domestic pigs. 8. These results underscore the limited utility of assessments of vasopressin receptor binding and vasopressin-stimulated adenylate cyclase activities in vitro to predict functional antidiuretic activities in vivo. PMID- 8224730 TI - Interaction between inhalational anaesthetics and enoximone on isolated heart muscle. AB - 1. The present study describes the effects of halothane or isoflurane on enoximone activity in the isolated left atria of the rat. 2. Concentration response curves were obtained for the positive inotropic effects of enoximone on electrically stimulated left atria. 3. Enoximone significantly (P < 0.01) increased the contractile force (56% maximum) with all the concentrations tested (10(-9) -10(-3) M). 4. When halothane (1.5% v/v) was present in the organ bath, the maximum effect obtained with enoximone (9%) was significantly lower than that obtained with enoximone alone. 5. Similar results were obtained with enoximone in the presence of halothane plus diltiazem. Isoflurane (1.5% v/v) did not significantly modify the maximum effect obtained with enoximone alone. 6. The administration of diltiazem antagonized the positive inotropic effects of enoximone in the presence of isoflurane or halothane. 7. These results shows that halothane, but not isoflurane, decreased the potency of enoximone on the isolated left atria and suggests that this effect may be mediated by the blocking of the influx of extracellular calcium through voltage-dependent calcium channels inhibited by diltiazem. PMID- 8224731 TI - "Selectivity" and "specificity": what do they mean? PMID- 8224732 TI - Multiple neuropeptide Y receptors are involved in cardiovascular regulation. Peripheral and central mechanisms. AB - 1. Neuropeptide Y (NPY) occurs in both the central and peripheral nervous system. In the periphery, NPY coexists with noradrenaline (NA) in perivascular sympathetic fibers. 2. NPY has a vasopressor effect, reflecting direct vasoconstriction of blood vessels and potentiation of the NA-evoked response. NPY also suppresses the release of NA from sympathetic fibers. 3. The post- and pre junctional NPY receptors are referred to as Y1 and Y2, respectively. They recognize not only NPY but also the homologous gut hormone peptide YY (PYY). 4. The Y1 and Y2 receptors have been characterized in numerous test systems using analogs of NPY/PYY. Already the deletion of the first N-terminal amino acid (NPY 2-36) results in a marked loss of potency at the Y1 receptor. The Y2 receptor is much less dependent upon an intact N-terminus, and a wide range of C-terminal NPY fragments retain quite high potency. 5. Recently, yet another NPY receptor, Y3, that is distinct from Y1 and Y2 in that it recognizes PYY poorly, has been demonstrated in the brainstem and in the periphery. 6. Further attempts to characterize the various receptor types have relied on truncated and substituted analogs of NPY/PYY. Although such studies suggest the existence of at least three types of NPY receptors, the lack of antagonists has represented a problem. 7. Since NPY may regulate cardiovascular functions via peripheral and central receptors its physiological and possibly pathophysiological significance has attracted much attention. 8. The responsiveness to NPY seems to be altered in animal models of hypertension and elevated plasma levels of NPY have been found in patients under various conditions of stress and in primary hypertension. A number of studies have suggested that NPY may be a pathogenetic factor behind primary hypertension. 9. Antagonists for the various NPY receptors would be useful for an analysis of which effects of these peptides are physiologically relevant. It is tempting to predict that both agonists and antagonists of the NPY receptors could be useful as drugs, for instance, in the treatment of primary hypertension. PMID- 8224733 TI - Physiological mechanisms and pharmacological regulation of acute collateral perfusion. AB - 1. Pre-existing collateral vessels provide an alternative route for blood flow following acute arterial occlusion. 2. In a novel model of acute arterial occlusion in the isolated rabbit ear vascular bed the development and maintenance of collateral perfusion has an obligatory dependence on endothelium-derived relaxing factor (EDRF) activity. 3. This endothelium-dependent mechanism is severely impaired in dietary-induced hypercholesterolaemia, leading to substantially reduced collateral perfusion. 4. Collateral perfusion may also be influenced by vasodilators, specifically in the rabbit ear model the potassium channel opener BRL 38227 greatly enhances perfusion, sodium nitroprusside (an exogenous analogue of EDRF) has early but limited beneficial effects while verapamil is without effect. PMID- 8224734 TI - Hepoxilins. AB - 1. This article reviews the formation, metabolism and pharmacological actions of the hepoxilins. These are biologically active hydroxy epoxide derivatives of arachidonic acid formed through the 12-lipoxygenase pathway. 2. This review summarizes literature data available at the time of writing of this article. PMID- 8224735 TI - The evidence for the existence of cannabinoid receptors. PMID- 8224736 TI - Subtypes of NMDA receptors. AB - 1. Beginning with electrophysiological evidence for two populations of receptors for N-methyl-D-aspartate (NMDA) which did or did not respond to the agonist quinolinic acid, evidence has grown for such subdivision. 2. Data from binding studies is consistent with differences between three NMDA receptors in the striatum, thalamus and cerebellum with respect to their preferences for agonist or antagonist binding and the modulation of binding by dizocilpine, cations and polyamines. 3. The recent isolation and sequencing of several different molecular species of NMDA receptor supports the view that at least two pharmacologically distinct sites exist, with the cerebellar receptor being unique in the brain. PMID- 8224737 TI - The L-arginine/nitric oxide pathway and non-adrenergic, non-cholinergic relaxation of the lower urinary tract. PMID- 8224738 TI - Treatment of muscle damage, induced by high intracellular Ca2+, with calmodulin antagonists. AB - 1. Incubation of rat diaphragm muscles in the presence of Ca(2+)-ionophore A23187, which causes accumulation of free intracellular Ca2+, induced severe myofibrils damage. Electron microscopic studies have revealed that calmodulin (CaM) antagonists, trifluoperazine, thioridazine, pimozide and CGS 9343B, were most effective in preserving muscle structure. 2. The CaM antagonists raised the decreased glucose-1,6-bisphosphate levels, induced by high Ca2+, with a concomitant activation of the reduced cytosolic phosphofructokinase (the rate limiting enzyme of glycolysis) and thereby cytosolic glycolysis. 3. All four CaM inhibitors also prevented solubilization of cytoskeleton-bound glycolytic enzymes by high Ca2+. 4. The protective effect of these compounds on cytosolic and cytoskeletal glycolysis, was also expressed by their action in preserving muscle ATP levels. 5. The present experiments suggest that CaM antagonists may be effective drugs in treatment of muscle damage and various muscle diseases, which are characterized by a high pathological increase in intracellular Ca2+. PMID- 8224739 TI - Vasodilatory effects of flavonoids in rat aortic smooth muscle. Structure activity relationships. AB - 1. Flavonoids relaxed the contractions induced by noradrenaline, KCl or phorbol 12-myristate, 13-acetate in rat aortic strips, the order of potency being: flavonols (quercetin, kaempferol, pentamethylquercetin) > flavones(luteolin, apigenin) > flavanols((+)-catechin, (-)-epicatechin) which correlates with the reported order of potency to inhibit protein kinase C. 2. The relaxant effects of kaempferol and luteolin were slightly potentiated by isoprenaline and those of pentamethylquercetin, kaempferol and apigenin by sodium nitroprusside. 3. It is concluded that the main vasodilatory mechanism of flavonoids seems to be the inhibition of protein kinase C. Inhibition of cyclic nucleotide phosphodiesterases or decreased Ca2+ uptake may also contribute to their vasodilatory effects. PMID- 8224740 TI - Chronic effects of SQ29,852, a new angiotensin converting enzyme (ACE) inhibitor with a phosphonic acid group, in experimental hypertensive rats. AB - 1. Chronic effects on blood pressure of SQ29,852 administered daily for 21 consecutive days were compared to those of captopril and enalapril in spontaneously hypertensive (SHR) and two-kidney-one-clip renal hypertensive (CLIP) rats. 2. SQ29,852 showed significant chronic anti-hypertensive effects at 30 mg/kg in SHR and at 10 mg/kg in CLIP rats. Chronic anti-hypertensive effects of SQ29,852 were in-between those of enalapril and captopril. 3. Significant anti hypertensive effects lasted in enalapril-treated SHR for 7 days after discontinuation of treatment, while only a tendency toward persistence of anti hypertensive effects was noted in all animals treated with SQ29,852 and captopril. PMID- 8224741 TI - Pentachlorophenol (PCP) inhibits ion transport in the isolated toad cornea. AB - 1. Active chloride transport from the stroma to the epithelial surface (tear side) accounts for 80% of the amphibian cornea short-circuit current (SCC). 2. The effect of pentachlorophenol (PCP, a wood preservative) on the bioelectric parameters of the toad Caudiverbera caudiverbera isolated cornea was studied. 3. PCP applied to the epithelial surface in the concentration range 0.3-4.3 microM caused a dose-dependent inhibition of the PD and of the SCC in 7 corneas. This inhibition was irreversible at all concentrations after several washouts. The agent had no effect when applied to the endothelial surface. 4. In 4 experiments the inhibitory effect was partly reversed by the addition of 1 microM calcium ionophore A-23187 to the epithelial surface. 5. It is concluded that PCP is an inhibitor of corneal active chloride transport and that this structure shows greater sensitivity to this agent than other tissues. PMID- 8224742 TI - Involvement of catecholamines in the effect of U-50,488H on isolated left atria of the rat. AB - 1. The present study examined the effect of U-50,488H on auricular contraction of isolated left atria of the rat. 2. The negative inotropic action induced by the kappa-agonist was antagonized in the presence of propranolol (10(-8) or 5 x 10( 8) M), yohimbine (5 x 10(-7) or 10(-6) M) or in reserpinized rats (5 mg/kg i.p. 24 hr before the experiments). 3. These results suggest that catecholaminergic mechanisms are involved in the cardiac depressant effect induced by U-50,488H. PMID- 8224743 TI - Molecular requirement for anticonvulsant activity in a series of thiazolo-1,4 benzodiazepine derivatives and comparison with classical benzodiazepines. AB - 1. The behavioural and anticonvulsant effects of several thiazolo[3,2 d][1,4]benzodiazepines (TBZ) were studied after intraperitoneal administration in DBA/2 mice, a strain genetically susceptible to sound-induced seizures. 2. Anticonvulsant effects on seizures evoked by means of auditory stimulation (109 dB, 12-16 kHz) were evaluated in DBA/2 mice placed singly under a perspex dome. 3. Hypothermic activity was observed after the highest doses of the benzodiazepines studied. 4. In addition, some TBZ were examined for anticonvulsant properties with respect to clonus induced by pentylenetetrazol. 5. Our study demonstrated that some thiazolobenzodiazepine derivatives were more potent than clobazam, desmethylclobazam and chlordiazepoxide, and less potent than diazepam, desmethyldiazepam and alprazolam. 6. In the series of tricyclic benzodiazepines, thiazole nucleus fusion to the "d" edge of the 7-membered ring results in an effective increase of the energy barrier for the heptatomic system reversal, and is probably responsible for, jointly with the lack of C=N double bonds, lower activity with respect to the 1,4-benzodiazepine precursors. 7. The potency of various thiazolobenzodiazepine derivatives as inhibitors of specific [3H]flunitrazepam binding to membranes from cerebellum or hippocampus was evaluated. 8. All tested compounds produced concentration-dependent inhibition of [3H]flunitrazepam binding. 9. The pharmacological activity of TBZ2, the most active compound of this series, was significantly reduced by treatment with flumazenil (2.5 mg/kg i.p.), suggesting clear involvement of a benzodiazepine mechanism in the anticonvulsant activity of these compounds. PMID- 8224744 TI - Hypotensive drugs, sodium nitroprusside and prazosin inhibit the metabolic rate in rabbits. AB - 1. Thermoregulatory responses (metabolic rate, rectal and ear skin temperatures, respiratory rate) were measured after treatment with two antihypertensive drugs, i.e. when sodium nitroprusside (6 mg/kg/2 hr) or prazosin (0.75 mg/kg/3 hr) were applied to normothermic or febrile rabbits under thermoneutral conditions (19 +/- 1 degree C). 2. Both drugs significantly decreased blood pressure and metabolic rate. 3. These changes were accompanied by antipyresis. 4. In the case of normothermic rabbits only sodium nitroprusside produced prominent falls in body temperature. 5. The relationship between changes in blood pressure and the intensity of metabolic rate is discussed. PMID- 8224745 TI - Adenosine modulates the antinociceptive action of benzodiazepines. AB - 1. In the present study, adenosine modulation on the antinociceptive action of benzodiazepines (BZD) using the writhing test in mice was evaluated. 2. The i.c.v. BZDs tested (diazepam, midazolam and lorazepam) induced a dose-dependent antinociceptive action, that was not antagonized either by naloxone or by aminophylline. 3. The i.p. administration of adenosine-related compounds also produced a dose-dependent reduction in the number of writhings in mice. These effects were not antagonized by i.p. injection of naloxone but were antagonized by i.p. aminophylline. 4. The antinociceptive effect of BZDs was significantly enhanced by the administration of adenosine compounds, but this increased response was not modified by naloxone or by aminophylline. 5. The present findings could be explained by the fact that BZDs and adenosine-related compounds may interact in an additive manner, since the effects of these drugs may be due to a common mechanism of action or a common pathway. PMID- 8224746 TI - Ampicillin concentrations in radicular cysts following a single oral administration of bacampicillin. AB - 1. Ampicillin concentrations in cyst wall (wall) and cyst fluid (fluid) of radicular cyst and serum following a single oral administration of bacampicillin (equivalent to 500 mg of ampicillin) were measured by a paper disk method. 2. The mean peak concentrations of ampicillin in wall, fluid, and serum occurred at identical times, 1.5 hr, and were 2.39 micrograms/g, 0.77, and 10.24 micrograms/ml, respectively. 3. Mean ampicillin concentration ratios of wall/serum, fluid/serum, and fluid/wall at the peak time were 0.23, 0.07, and 0.40, respectively. 4. Mean ampicillin concentrations in wall and fluid at the peak time exceeded MIC (minimum inhibitory concentration) for 90% (0.5 microgram/ml) for clinically isolated strains of alpha-hemolytic Streptococci. PMID- 8224747 TI - Isolation and characterization of the substance isolated from Streptosporangium species which inhibits lactic acid production by oral bacteria. AB - 1. The tentatively named MLI, which inhibits lactic acid production in mutants streptococci, was isolated from Streptosporangium species and purified by solvent extraction followed by chromatography, and then powdered. 2. MLI greatly inhibited pH-decline and lactic acid production in glucose-containing reaction mixture by mutans streptococci or Lactobacillus species. 3. These inhibitory mechanisms were not caused by an anti-bacterial effect, since the addition of a sufficient concentration of MLI to inhibit lactic acid production to the culture, recovery of cells in culture was not affected, excluding L. acidophilus. 4. MLI directly affected lactate dehydrogenase (LDH) activity. 5. These results indicate that MLI may represent a new anti-cariogenic substance. PMID- 8224748 TI - Effects of adenosine agonists and antagonists on pulmonary ventilation in conscious rats. AB - 1. The effects of intravenous injections of adenosine agonists and antagonists on pulmonary ventilation were investigated in conscious male Sprague-Dawley rats. 2. Adenosine agonists reduced pulmonary ventilation, whereas an adenosine antagonist increased pulmonary ventilation. 3. The adenosine-induced decrease in ventilation could be partially blocked by pretreatment with either an adenosine or opioid antagonist and completely blocked by a combined pretreatment with adenosine and opioid antagonists. 4. Thus, endogenous adenosine appears to have an inhibitory role in the control of pulmonary ventilation in conscious rats, and the mechanism of action appears to involve both adenosine and opioid receptors. PMID- 8224749 TI - Biphasic GABA-A receptor-mediated effect on the spontaneous activity of the circular layer in cat terminal ileum. AB - 1. The GABA and GABA-A receptor agonist muscimol changed the spontaneous mechanical activity of a circular layer isolated from cat terminal ileum, while the selective GABA-B receptor agonist (+/-)baclofen had no effect. 2. GABA at doses ranging from 1 microM to 2 mM elicited concentration-dependent biphasic responses which consisted of a relaxation followed by contraction, with a tonic and a phasic component. The EC50 values, calculated at 95% confidence limits (CL), were 94.9 microM (83.5-109.8 microM) and 66.0 microM (51.2-75.5 microM) for the relaxation and contractile phases, respectively. 3. The GABA-induced biphasic responses were sensitive to bicuculline and picrotoxinin and were entirely mimicked by muscimol. Bicuculline competitively antagonized the effects of GABA and gave closely similar pA2 values for both phases of these responses- inhibitory and stimulatory. Cross-desensitization occurred only between GABA and muscimol and not between (+/-)baclofen and GABA, or (+/-)baclofen and muscimol. 4. Both bicuculline-sensitive phases evoked by GABA and muscimol were abolished by tetrodotoxin or atropine, but were unaffected by guanethidine or naloxone. 5. The present results suggested that the biphasic GABA effect on the mechanical activity of the circular layer in cat terminal ileum was mediated by prejunctional GABA-A receptors, most probably through an action on the cholinergic pathway. PMID- 8224750 TI - Calcium channel blockers apparently decrease noradrenaline release from nerve skin terminals in Caudiverbera caudiverbera. AB - 1. The effects of three calcium channel blockers, verapamil, diltiazem and nifedipine were examined on the response of the skin neuroepithelial synapse of the toad Caudiverbera caudiverbera to electrical stimulation. 2. The stimulus induced a significant rise in potential difference (PD) and short-circuit current (SCC). The three calcium antagonists reduced the responses in a dose-dependent manner. The greatest reduction was induced by verapamil followed by diltiazem and nifedipine. 3. Amiloride treatment did not affect the responses to electrical stimulation, indicating that the response to nerve stimulation is not due to current flowing through sodium channels. 4. When the preparation was blocked by either of the three antagonists, the skin response to noradrenaline was not affected. 5. It may be concluded that verapamil, diltiazem or nifedipine reduce the release of noradrenaline at the neuroepithelial synapse of C. caudiverbera. PMID- 8224751 TI - Endothelium-dependent vasodilator effects of the extract from Salviae Miltiorrhizae radix. A study on the identification of lithospermic acid B in the extracts. AB - 1. The aqueous extract of Salviae Miltiorrhizae radix (Chinese crude drug named "dan-shen") relaxed the noradrenaline-precontracted aorta with endothelium. 2. Vasodilation by the extract disappeared in aorta without endothelium, and was inhibited by pretreatment with 10(-4) M NG-monomethyl-L-arginine (L-NMMA) or 10( 5) M methylene blue. 3. The inhibition of the extract-induced vasodilation by L NMMA was reversed by L-arginine (3 x 10(-4) M). 4. The component of the extract was analyzed by chromatography, fast atom bombardment mass spectroscopy (FAB-MS) and 1H-NMR. 5. An active component of the extract, which showed endothelium dependent vasodilation, was found to be identical with lithospermic acid B. PMID- 8224752 TI - Effects of non-opioid antitussives on hypoxia-induced electrical changes in rat hippocampal slices: a comparative study with anticonvulsant drugs. AB - 1. The effects of the non-opioid antitussives caramiphen and carbetapentane and of the anticonvulsants 5,5-diphenylhydantoin and MK 801 were tested towards hypoxia-induced electrical changes in rat hippocampal slices. 2. The incidence of appearance of hypoxia-induced epileptiform bursting was significantly decreased (P < 0.05) by carbetapentane (50-100 microM), caramiphen (50-100 microM), 5,5 diphenylhydantoin (25-50 microM), and the glutamate antagonist dizocilpine (MK 801, 25-50 microM). 3. The incidence of reappearance of the CA1 population spike after hypoxia was significantly increased (P < 0.05) by carbentapentane (50-100 microM), caramiphen (50-100 microM), 5,5-diphenylhydantoin (25-50 microM), and MK 801 (25-50 microM). 4. The results suggest a useful role for non-opioid antitussives and some anticonvulsants in the treatment of hypoxia-induced functional disturbances. PMID- 8224753 TI - Studies on the hypocalcemic actions of salmon calcitonin and ultimobranchial gland extracts in the freshwater teleost Cyprinus carpio. AB - A single injection of synthetic salmon calcitonin (sCT) or homologous ultimobranchial gland (UBG) extract caused a significant reduction in total and ultrafiltrable plasma Ca2+ in a freshwater teleost, Cyprinus carpio. A time-bound analysis on the effect of sCT or UBG extract caused a short-duration hypocalcemia and the response was more pronounced in fish kept in low-Ca2+ water. Sexually immature fish showed a greater response than did mature fish. In all the studies the protein-bound Ca2+ fraction remained unaffected. The observation thus indicate a potent hypocalcemic role of calcitonin or UBG in freshwater common carp under laboratory conditions. PMID- 8224754 TI - Subcellular action of estradiol-17 beta in a freshwater prawn Macrobrachium rosenbergii. AB - Injections with different doses (0.1, 0.25, 0.5, 1.0, 2.0, and 4.0 micrograms/g) of estradiol-17 beta (E2), administered three days consecutively, showed a statistically significant increase in mitochondrial Na(+)-K(+)-ATPase, cytosolic malate dehydrogenase, and cytosolic glucose-6-phosphate dehydrogenase activities in a dose-dependent manner in the hepatopancreas of the freshwater prawn (Macrobrachium rosenbergii) on the 4th day of treatment compared to the control values. A lower dose of 0.05 microgram/g was without any effect on these enzyme activities. A uniform increase in the Mg(2+)-ATPase activity was observed after injections with 0.5, 1.0, and 2.0 micrograms/g of E2. Ergosterol, a nonsex steroid did not show any change in the malate dehydrogenase and glucose-6 phosphate dehydrogenase activities on which this compound was tested at a 2.0 micrograms/g dose, compared to the control values. Simultaneous injection of tamoxifen (0.5 microgram/g), an antiestrogenic compound, with E2 (2 micrograms/g) caused inhibition of the E2-induced rise in mitochondrial Na(+)-K(+)-ATPase and cytosolic NADP-linked malate dehydrogenase activities. Conversely, tamoxifen (0.5 and 1.0 microgram/g) behaved as an estrogen agonist to the response (increase) of Mg(2+)-ATPase and cytosolic glucose-6-phosphate dehydrogenase activities. Potentiation of the estrogen effect with tamoxifen (1.0 microgram/g) was observed in these enzyme activities when used simultaneously with E2 (2 micrograms/g). Use of cycloheximide (0.5 mg/liter), a protein synthesis blocker, inhibited the inhibited the E2 (2 micrograms/g)-induced increase in all the enzyme activities studied. The data show specific and prominent subcellular action of estrogen with an indication of its role in energy-dependent ion transport and metabolic activation in hepatopancreas of the freshwater prawn. PMID- 8224755 TI - Simple N-linked sugar chains are bound to the lutropin of the bullfrog rana catesbeiana. AB - The structures of Asn-linked sugar chains of the bullfrog (Rana catesbeiana) lutropin (LH) with different isoelectric points (pI 9.3 and pI 8.8) have been analyzed. The LH of pI 9.3, which is three times more active in Xenopus ovulation assays and radioreceptor assays with Xenopus testis than LH of pI 8.8, has Man GlcNAc-GlcNAc and Man-Man-GlcNAc-GlcNAc as the major sugar chain components both at Asn-57 and 83 of the alpha-subunit. On the other hand, the alpha-subunit of the LH of pI 8.8 contained more mannose and phosphate bound at the nonreducing end of the sugar chains. This is the first study to identify the presence of a phosphate group bound to the mannose in LH. The structure of the major sugar chain bound to the beta-subunit was Man-Man-GlcNAc-(Fuc)GlcNAc at Asn-8 of the LH from both pIs. These sugar chains bound to the bullfrog LH are more simple than those of mammalian LH, which have N-acetyl-galactosaminylated biantennary oligosaccharides and more complicated sugar chains. The microheterogeneous pI of bullfrog LH is thus due to the different phosphate contents in the sugar chains bound to the alpha-subunit instead of sulfate groups which are found in mammalian lutropins. PMID- 8224756 TI - Changes in morphogenetic hormone titers in isolated workers of the termite Reticulitermes flavipes (Kollar). AB - The daily titers of juvenile hormone and ecdysteroids were determined for workers of the Eastern subterranean termite Reticulitermes flavipes (Kollar) (Isoptera, Rhinotermitidae) following isolation of the workers from soldiers and reproductives. Experiments have demonstrated that isolation leads to biochemical and physiological changes that result in a presolider molt. Juvenile hormone (JH) and molting hormone (MH) titers were determined in hemolymph samples using radioimmunoassay. The identity of the ecdysteroids as predominantly 20 hydroxyecdysone (90%) and ecdysone (10%) was determined by HPLC using pooled extracts of hemolymph. PMID- 8224757 TI - Excessive abundance and dynamics of unusual ecdysteroids in Pycnogonum litorale strom (Arthropoda, Pantopoda). AB - In embryos, larvae, and juveniles of Pycnogonum litorale, unusual ecdysteroids have been found at excessively high levels. Concentrations and dynamics of the following eight ecdysteroids were determined by HPLC: 20-hydroxyecdysone, 20 hydroxyecdysone 22-glycolate, (25R) and (25S) isomers of 20,26-dihydroxyecdysone 22-acetate, 22-deoxy-20,26-dihydroxyecdysone, 20-hydroxyecdysone 22-acetate, 22 deoxy-20-hydroxyecdysone, and ecdysone 22-glycolate. The excessively high content of ecdysteroids is found in all developmental stages. Juvenile pycnogonids contain the highest total ecdysteroid amounts ever found in arthropods: 0.1% of their body dry weight. 20-Hydroxyecdysone 22-acetate is the prominent ecdysteroid and represents at all stages of both sexes, 66.2-85.8% of the total ecdysteroids. This compound, as well as all other ecdysteroids without a free 22-OH group, are presumed to be inactive as molting hormones. In contrast to these compounds, 20 hydroxyecdysone is present only in amounts similar to those in other arthropods. Furthermore, it is the only ecdysteroid with its peak at apolysis. It is regarded as the molt-promoting hormone. The origin and function of the other ecdysteroids in different developmental stages are discussed. PMID- 8224758 TI - Effect of estrogen in relation to dietary vitamin D3 and calcium on activity of intestinal alkaline phosphatase and Ca-ATPase in immature chicks. AB - The interaction between 17 beta-estradiol (E2), vitamin D3 (D3), and dietary Ca on the activities of Ca-ATPase and alkaline phosphatase (AP) was determined in the intestine of young female chicks. Chicks (n = 36) were assigned to two groups, one of which was transferred to a low Ca (0.2%) diet and the other maintained on a regular diet. One week later, each group was further divided into three subgroups and given daily injections of 0(oil), 0.25, or 0.5 mg E2/kg body wt for 14 days. E2 treatment as well as low dietary Ca significantly increased AP activity (P < 0.05), whereas the highest E2 dose decreased jejunal Ca-ATPase (P < 0.05). In a separate study, day-old chicks (n = 40) fed a purified diet supplemented with or without D3 for 24 days were divided into two subgroups and administered daily injections of either 0 or 0.25 mg estrogen 3-benzoate/kg body wt for 5 days. E2 alone or in combination with D3 failed to change Ca-ATPase activity in either the duodenum or the jejunum. However, E2 enhanced the D3 stimulated AP activity measured in the supernatant of duodenum (D3, P < 0.001; E2, P > 0.05; E2 x D3, P < 0.05) and jejunum (D3, P < 0.001; E2, P > 0.05; E2 x D3, P = 0.06). Daily injections of 0.5 mg E2/kg body wt for 6 days to 6-week-old D3-adequate chicks (n = 16) significantly increased AP activity in jejunum but not in liver and kidney (P < 0.05). In conclusion, E2 treatment enhanced the activity of intestinal AP but not Ca-ATPase. This enhancement was independent of dietary Ca, but was D3-dependent and tissue specific. The results suggest that the pubertal increase in plasma E2 can affect Ca absorption from the intestine by increasing the activity of AP. PMID- 8224759 TI - In vivo GnRH responsiveness of LH secretion in the female turtle, Trachemys scripta, in relation to the reproductive stage. AB - The effects of several forms of gonadotropin-releasing hormone (GnRH) on the in vivo secretion of luteinizing hormone (LH) were investigated in the female turtle Trachemys scripta. Serial injections of large vitellogenic turtles with 1-2 micrograms (ca. 0.15 micrograms/100 g turtle) of chicken GnRH-I (cGnRH-I) and multiple blood sampling revealed that mature females were unresponsive to GnRH. In contrast, both forms of avian GnRH, cGnRH-I and cGnRH-II, and an avian GnRH analogue (GnRHa) significantly increased circulating levels of LH in previtellogenic turtles. In these turtles, plasma LH increased within 10 min after the initial injection of 0.5 microgram GnRHa or cGnRH-I, but this increase in LH was generally transient and could not be sustained for more than 2 hr even with repeated GnRH injections. Turtles treated with GnRHa on Day 1 showed attenuated pituitary responsiveness to subsequent GnRH challenges on Day 2, whereas turtles treated with both native forms of chicken GnRH remained equally responsive on Day 2. In vitro, GnRHa was more resistant to degradation in plasma than both native forms of GnRH, and the clearance rate of GnRHa in vivo was also slower than that of cGnRH-I. The results show, for the first time, that in vivo GnRH administration stimulates and desensitizes LH secretion in the turtles, but the GnRH responsiveness is related to reproductive status. Furthermore, the results suggest that the desensitizing effects of various forms of GnRH in vivo may be related to their clearance rate. PMID- 8224760 TI - Vitamin D receptors in a naturally vitamin D-deficient subterranean mammal, the naked mole rat (Heterocephalus glaber): biochemical characterization. AB - 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), the hormonally active metabolite of vitamin D3 interacts with its nuclear/cytosolic receptor to induce biological responses in target tissues. Naked mole rats appear to be naturally deficient in vitamin D. The questions arise whether these animals possess the 1,25(OH)2D3 receptor (VDR) and whether they are capable of responding to 1,25(OH)2D3 via receptor-mediated pathways. Various tissues (intestine, kidney, Harderian glands, and skin) were examined for the presence and biochemical characterization (as indicated by saturation, sucrose density gradient, DNA binding, and ligand competitive analysis) of VDRs. In addition homologous upregulation of VDRs in these tissues and induction of 25-hydroxyvitamin D3-24-hydroxylase (24-OHase) in the kidney was studied as indicators of the VDR-mediated biological responses. Naked mole rats have VDRs in the intestine, kidney, and Harderian gland but not in skin. Biochemical characterization of VDRs and VDR-mediated biological responses in the intestine and kidney correspond to those found in similar target tissues of other mammals. Harderian gland VDR is at a lower concentration yet shows a markedly higher affinity and selectivity to 1,25(OH)2D3 than that of the intestine and kidney. Vitamin D3 supplementation resulted in VDR upregulation in the intestine and kidney and induced renal 24-OHase but had no effects on VDRs in Harderian glands. These data demonstrate that naked mole rats possess VDRs in intestine, kidney, and Harderian glands; these VDRs differ in their biochemical characteristics. PMID- 8224761 TI - Isolation of prolactin and growth hormone from the pituitary of the holostean fish Amia calva. AB - Pituitaries from adult male and female Amia calva (Order Holostei) were acid extracted and fractionated by gel filtration column chromatography and reversed phase high performance liquid chromatography. This two-step isolation procedure yielded homogeneous pools of Amia prolaction (PRL) and growth hormone (GH). The amino acid composition of both purified polypeptides was determined. Primary sequence analysis of the first 22 positions at the N-terminal of Amia PRL revealed that this region has 63% sequence identity with eel PRL-1. The N terminal region of Amia PRL lacks the disulfide bridge which is characteristic of tetrapod PRLs. Primary sequence analysis of the first 24 positions at the N terminal of Amia GH revealed that this region has 62% sequence identity with eel GH and 54% sequence identity with both blue shark GH and sea turtle GH. Based on N-terminal analysis, it appears that Amia PRL and GH are more closely related to teleost PRLs and GHs than they are to tetrapod PRLs and GHs. PMID- 8224762 TI - Low temperature and short days together induce thyroid activation and suppression of LH release in Japanese quail. AB - We have found in Japanese quail that low ambient temperature is required to terminate breeding activity in the presence of short days (Wada et al., 1990, Gen. Comp. Endocrinol. 80, 465-472; Tsuyoshi and Wada, 1992, Gen. Comp. Endocrinol. 85, 424-429). To elucidate the mechanism for photoperiodic and temperature regulation of the release of luteinizing hormone (LH), several serum variables were measured in three groups of mature male birds: (a) birds kept on long days of 16L:8D, 24 hr at 19 degrees as an initial control group (Group IC), (b) birds transferred from long days to short days of 8L:16D, 24 hr at 19 degrees (Group S) for 14 days, and (c) birds transferred to short days of 8L:16D and low temperature cycles of 12 hr, 19 degrees: 12 hr, 9 degrees (Group SL) for 14 days. Testicular mass and plasma concentrations of LH significantly decreased to nonbreeding levels in Group SL, but not in Group S, confirming our previous results. Hematocrit, serum osmolarity, and concentrations of Na+, K+, Ca2+, Mg2+, and Cl- ions were not different among the three groups. Serum concentrations of free fatty acids were increased in Group SL, but the increase was not statistically significant. On the other hand, plasma concentrations of thyroid hormones changed significantly; thyroxine (T4), but not triiodothyronine (T3), increased in Group S, and T3, but not T4, increased in Group SL. To follow the changes in plasma levels of LH, T4, and T3 during the treatments, blood samples were collected every other day for 2 weeks from birds in the three groups of mature male birds described above.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224763 TI - Isolation, identification, and synthesis of AKH-I4-10 from Locusta migratoria. AB - A heptapeptide was isolated from brain-corpora cardiaca-corpora allata suboesophageal ganglion extracts of the locust, Locusta migratoria. Biological activity was monitored during HPLC purification by observing the myotropic effect of column fractions on the isolated hindgut of Leucophaea maderae. The primary structure of this myotropic peptide was established as: Phe-Thr-Pro-Asn-Trp-Gly Thr-NH2. The chromatographic and biological properties of the synthetic peptide were the same as those of the native peptide, thus confirming structural analysis. This heptapeptide is identical to the carboxyterminal heptamer of AKH-I and therefore designated as AKH-I4-10. AKH-I4-10 has no adipokinetic activity. AKH-I4-10 is most likely a breakdown product of Lom-AKH-I, suggesting that an endopeptidase which cleaves between Asn and Phe is present in the brain complex of L. migratoria. Such an endopeptidase has recently been characterized in in synaptic membranes of the nervous system of Schistocerca gregaria. PMID- 8224764 TI - Annual variations in hypothalamic serotonin and monoamine oxidase in the catfish Heteropneustes fossilis with a note on brain regional differences of day-night variations in gonadal preparatory phase. AB - In Heteropneustes fossilis, significant annual variations in serotonin (5-HT) and monoamine oxidase (MAO) were found in the hypothalamus both at 12 and 24 hr with high values of content and turnover index (TI) during gonadal recrudescence, and low values during gonadal quiescence. The 5-HT content reached high levels during April-July (late preparatory, prespawning, and spawning phases). The TI of 5-HT showed two peaks in its midday value; a major peak occurred in March (midpreparatory phase) and a minor one in June (prespawning phase). High MAO activity was seen in June (prespawning phase) and low values during February, March, and April (preparatory phase). Significant day-night variations in 5-HT content and TI, and MAO activity were observed in the hypothalamus and telencephalon in February and March (early and midpreparatory phase), but not at other times of the year. In the whole brain and separate regions such as the thalamus, midbrain, and hindbrain no day-night patterns were seen. A comparison of the data shows that the increase in 5-HT content and TI from February to March was significantly higher in the hypothalamus than in telencephalon and the opposite is true for MAO. The results suggest that the high hypothalamic activities of 5-HT and MAO during recrudescence is related to breeding activity; the day-night variations during the early and midpreparatory phase in these variables may be related to initiation of breeding in this species. PMID- 8224765 TI - Epidermal growth factor stimulates protein synthesis in primary cultures of salmon hepatocytes. PMID- 8224766 TI - Receptors for atrial natriuretic factor (ANF) in kidney and adrenal tissue of urodeles--lack of angiotensin II (A II) receptors in these tissues. AB - Binding sites for atrial natriuretic factor (ANF) and angiotensin II (A II) were localized and quantified in renal and adrenal tissue of the Urodele Ambystoma mexicanum by quantitative in vitro autoradiography using 125I-ratANF(99-126) and 125I-[Val5]-A II or 125I-human A II as labeled ligands. Specific 125I-ratANF(99 126) binding was present in glomeruli, renal tubules, and adrenal tissue. ANF had heterogeneous binding sites exhibiting positive cooperativity with a half maximal binding concentration (EC50) of 124 +/- 12 pM in glomeruli and 118 +/- 18 pM in adrenal tissue (n = 8). The corresponding maximal binding capacities (Bmax) were 0.84 +/- 0.10 and 0.72 +/- 0.18 femtomol/mm2. 125I-ratANF(99-126) binding was competitively displaced by unlabeled ANF analogues with an intact disulfide bridge showing a lower affinity than the iodinated ligand. In in vitro perifusion experiments ANF inhibited basal as well as ACTH-stimulated steroid secretion whereas catecholamine release was not affected indicating that ANF binding in adrenal tissue is restricted to steroidogenic interrenal cells. In vivo experiments demonstrated that during acclimation to 1.5% salt water made using seawater salt the number of ANF binding sites in kidney glomeruli decreased after 12 hr and 7 days while no significant changes were observed in adrenal tissue. No A II binding sites were detected either in renal or adrenal tissue indicating a lack of A II receptors in these tissues, which was confirmed by in vitro perifusion experiments which showed that A II did not affect adrenal steroid and catecholamine secretion. PMID- 8224768 TI - Intestinal 5'deiodinase activity of developing and adult chickens selected for high and low body weight. AB - Intestinal 5'deiodinase activity (5'D) in adults from lines of chickens selected for high (HW) and low (LW) juvenile body weight was studied in vitro using reverse T3 substrate. This avian intestinal 5'D activity is similar in its biochemical characteristics to the hepatic 5'D activity of mammals and birds. Adult 5'D activity was higher in line HW than LW; a pattern opposite to that prior to hatching. Previous studies have shown that thyroid hormones are important in triggering differentiation of the intestine during the latter part of embryonic life. Our studies indicate that the intestinal tissue is capable of producing T3 that may be important in stimulating differentiation of the intestine itself, in preparation for posthatching food assimilation. The intestines of LW embryos, which hatch earlier than HW embryos, show both more rapid maturation, as indicated by intestinal alkaline phosphatase activity (a marker of differentiation), and higher 5'D activity. Increases in intestinal 5'D in the HW line at hatch may be important in the differential intestinal growth that leads to higher relative intestinal weights in HW than LW chicks after hatch. The magnitude of 5'D activity and the relative size of the intestine suggest that this organ could be an important contributor to plasma T3. PMID- 8224767 TI - Metabolism of exogenous steroids by anuran larvae. AB - Uptake and metabolism of exogenous steroids [testosterone (T), estradiol (E2), and corticosterone (B)] were examined in anuran larvae. Radioimmunoassay (RIA) of steroids dissolved in water revealed that as much as 0.03 mg of T or E2 were cleared from water by a single Xenopus laevis tadpole in 24 hr. Studies using RIA in combination with counting of total [3H]steroids suggested that clearance was mainly due to metabolism and not to uptake and storage by the tadpole. Thin-layer chromatography and HPLC analysis confirmed metabolism and revealed that T was metabolized to at least three ether-extractable compounds as well as to water soluble compounds (nonextractable in ether) by nine anuran species, representing eight families. Most species metabolized E2 to nonextractable compounds, but some produced organic metabolites. In contrast, B was not taken up, cleared, or metabolized by any of the nine species. Studies in X. laevis using [3H]T and radioinert T in the same container confirmed that the decrease in radioimmunoassayable T was due to metabolism as determined by TLC. In a separate study, [3H]T was metabolized more extensively when injected directly into the tadpole than when dissolved in water. Additional studies with X. laevis revealed that steroids were also metabolized in tadpole-conditioned water (water exposed to tadpoles), but the metabolites produced represented < 10% of those produced when tadpoles were present. This metabolism was confined to particulate matter from the gut of tadpoles. Metabolites extracted from tadpoles were similar to metabolites found in the water. PMID- 8224769 TI - Pharmacokinetics of atrial natriuretic peptide in Pekin ducks. AB - To quantify the pharmacokinetics of atrial natriuretic peptide (ANP) in the blood of Pekin ducks, synthetic avian ANP was intravenously infused at various rates and the associated plasma concentrations of immunoreactive ANP (irANP) were measured by radioimmunoassay. At rates of 10, 25, 50, 100, and 200 ng/kg/min, steady-state arterial plasma levels, achieved within 10 min, showed a linear dose plasma level-response relationship which indicated a mean plasma clearance rate of 174.4 +/- 5.4 ml/kg/min. After stopping the 200 ng/kg/min infusion of ANP, the decay curve of plasma irANP was consistent with first-order kinetics, with the peptide having an elimination half-life time of 1.2 min. It is concluded that ANP is rapidly removed from the circulation of Pekin ducks. PMID- 8224770 TI - Solubilization and purification of the gonadotropin (GTH II) receptor from rainbow trout (Oncorhynchus mykiss) ovaries. AB - The rainbow trout (Oncorhynchus mykiss) ovarian gonadotropin (GTH II) receptor was solubilized by extraction with the nonionic detergent 1% Triton X-100 in the presence of 20% glycerol. The hormone-binding characteristics of the soluble receptors were similar to those of membrane-bound receptors: the Scatchard plot of the equilibrium binding data produced a straight line, suggesting that the solubilized GTH II receptors, like membrane-bound receptors, contained a single class of high affinity 125I-sGTH II binding sites with an association constant of 2-5 x 10(10) M-1 (Ka = 1.4-2 x 10(10) M-1 for membrane-bound receptor). The maximal binding capacity was very low and varied from 7 to 17 fmol/mg proteins (about 5 fmol/mg ovarian membrane protein). The soluble receptor was purified by a simple and rapid immunoaffinity chromatography. The sGTH II-solubilized receptor complex was adsorbed to anti-sGTH II beta-subunit gammaglobulins covalently linked to Sepharose 4B and then eluted with an acidic buffer. About 50% of the binding activity present in the Triton X-100 extract was recovered in the pH 4 eluate. The other binding sites were eluted as a hormone-receptor complex and/or a damaged form. The free purified receptor presented a Ka of 1.3 x 10(10) M-1 in agreement with those found in membrane preparation and solubilized extract. PMID- 8224771 TI - A second glucagon in the pancreatic islets of the daddy sculpin Cottus scorpius. AB - The peptide hormone glucagon has been isolated from the islet tissue (Brockmann bodies) of the teleost Cottus scorpius (daddy sculpin) and sequenced. The sequence is HSEGTSNDYSKYLEDRKAQDFVQWLMNN differing at four positions from the glucagon found earlier in the same species by Conlon and coworkers (1987b, Eur. J. Biochem, 164, 117-122). Thus sculpin, in common with anglerfish, possesses two distinct glucagons. Comparative sequence data are presented as a phylogenetic tree. PMID- 8224772 TI - Sexual activity and plasma levels of sex steroids in the aspic viper Vipera aspis L. (Reptilia, Viperidae). AB - Reproductive behavior and associated sexual activity was studied in individual male and female Vipera aspis over a 3-year period in western France in an attempt to correlate mating behavior with blood levels of gonadal sex steroids. Males had higher average levels of both testosterone (T) and 5 alpha-dihydrotestosterone (DHT) than females. Levels of progesterone (P) did not differ significantly between the two sexes but estradiol-17 beta (E2) concentrations were significantly higher in females during the season of mating. Spring mating behavior and copulation in males was associated with significantly increased levels of T and DHT, compared with postmating males; and a similar, but not significant trend, was evident with autumnal mating. The only statistically significant hormonal difference detected in males showing no sexual activity in autumn, was an elevated level of E2 at 0.52 +/- 0.20 ng/ml compared with 0.09 +/- 0.03 ng/ml in spring-breeding males (P = 0.05). Estrus in females is associated with increased levels of all four steroids but significant only for E2 and DHT. Levels of P were significantly reduced in females displaying seasonal anestrous in the spring immediately following reproduction. Females not displaying estrus in either spring or autumn had significantly lower plasma DHT and E2. Although mating behavior in males is associated statistically with elevated levels of T and DHT, a tight correlation is not obvious at the individual level, suggesting that increased concentrations of androgens are a necessary, but not sufficient, condition for mating. The data from V. aspis suggest that, as in a number of other reptilian species, high circulating levels of androgens function to "condition" or "organize" sexual behavior in males which may be displayed at some later time, well after actual levels have fallen, thus engendering the impression that reproductive behavior may be temporally dissociated from essential hormonal stimuli. PMID- 8224773 TI - Activation of somatolactin cells in the pituitary of the rainbow trout Oncorhynchus mykiss by low environmental calcium. AB - The effects of changes in environmental calcium on the activity of hypophysial somatolactin (SL) cells were examined in the rainbow trout. In fish 10 and 21 days after transfer from fresh water (FW) to FW containing 10 mM calcium (Ca-rich FW) or to 80% seawater (80% SW) containing 8 mM calcium, sectional nuclear areas of SL cells were reduced. The levels of SL-mRNA, determined by in situ hybridization, were lower 10 days after transfer to Ca-rich FW. When the fish were transferred from Ca-rich FW to FW, nuclear areas of SL cells increased along the significant increases in SL-mRNA and plasma SL levels. The increased activity of SL cells in the fish in low calcium environment suggests a hypercalcemic action as one of the functions of SL. On the other hand, a significant reduction in prolactin cell activity occurred after transfer to 80% SW, but not after transfer to Ca-rich FW. Growth hormone cells were activated 21 days after acclimation to 80% SW, in accord with its described SW-adapting action in salmonids. PMID- 8224774 TI - Isolation and characterization of two forms of Xenopus prolactin. AB - Two forms of highly purified prolactin (PRL) were obtained from pituitary glands of Xenopus laevis by extraction of acetone-dried powder with acid acetone and high-performance liquid chromatography on anion exchange, gel filtration, and reverse-phase columns. Purification was monitored by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis employing antiserum against bullfrog PRL. The Xenopus prolactins (xPRL-I and xPRL-II) thus obtained were shown to have similar molecular weights of 23,000 as determined by SDS-PAGE. The isoelectric points of xPRL-I and xPRL-II determined by isoelectric focusing were 5.6 and 5.3, respectively. Both hormones blocked T4-induced shrinkage of Xenopus tadpole tail fin in vitro. The amino acid compositions of the xPRLs resembled that of bullfrog PRL. The partial amino acid sequences of xPRL-I and of xPRL-II showed 78 and 68% homology with the comparable portion of the sequence of bullfrog PRL, respectively. Homology between xPRL-I and xPRL-II was 90%. PMID- 8224775 TI - Synthesis of vitellogenin by eel (Anguilla anguilla L.) hepatocytes in primary culture: requirement of 17 beta-estradiol-priming. AB - Control and 17 beta-estradiol-primed eels were used to investigate the hormonal requirement of vitellogenesis in an immature fish, the eel. A primary culture of isolated liver cells from female silver eels was developed. The hepatocytes were maintained as monolayers on poly-L-lysine coated dishes for up to 12 days in a defined medium alone or supplemented with 17 beta-estradiol (E2, from 10(-8) to 10(-5) M). The amounts of vitellogenin (Vg) in the cells and secreted into the medium were measured at 2-day intervals using a homologous vitellogenin ELISA. Different E2-priming conditions were determined before hepatocyte isolation (one injection of 250 micrograms of E2 21 days, 17 days, or 24 hr). The vitellogenic response of hepatocytes to E2 stimulation was studied in relation to the duration of the E2-priming. After 8 days of culture, when hepatocytes from control eels were used, Vg was undetectable both in cells and in culture media, even if the culture was performed in the presence of E2 10(-5) M. However, Vg was detectable both in cells and in culture media of hepatocytes from E2-primed eels. If the priming was performed 24 hr before the culture, the Vg synthesis significantly increased (P < 0.001) in the presence of E2 10(-5) M after 10 days of culture but remained low. When the culture was performed 17 or 21 days after the priming, the level of the vitellogenic response was higher than after a short priming. In particular, with hepatocytes from 21-day E2-primed eel, the concentration of secreted Vg was 1.5 times higher than in control dishes (P < 0.01), in the presence of E2 10(-8) M after 12 days of culture. Higher doses of E2 (10(-5) M) increased Vg 2.7-fold over control values (P < 0.01) after 4 days of culture. In control dishes, cultured without steroid, the amounts of secreted and intracellular Vg remained unchanged over 12 days of culture (respectively, 72.8 +/- 2.7 ng/10(6) cells/48 hr and 28.7 +/- 2.7 ng/10(6) cells). These results show that cultured hepatocytes retain their functional capacity by synthesizing a specific protein, Vg, in the presence of E2 and there are dose- and time-related effects of E2 on in vitro Vg synthesis. The induction of hepatic vitellogenesis in vitro requires a preliminary in vivo E2-priming. PMID- 8224776 TI - Lungfish neurohypophysial hormones: chemical identification of mesotocin in the neurointermediate pituitary of the Australian lungfish Neoceratodus forsteri. AB - The neurohypophysial hormones contained in the neurointermediate lobe of the pituitary of the Australian lungfish Neoceratodus forsteri have been investigated. Mesotocin was identified by coelution with the synthetic peptide in high-pressure liquid chromatography, by Edman amino acid sequencing, by mass spectrometry, by C-terminal sequencing through carboxypeptidase Y, and cleavage with prolyl endopeptidase. Vasotocin, if present, would be in very small amounts and hydrins were not detected. Oxytocin appears absent. Although Neoceratodus and Protopterus have different habitats, the former being permanently aquatic, the latter burrowing during estivation, the proportions of neurohypophysial hormones stored in neurohypophysis were roughly similar in the two species and not apparently affected by environmental conditions. PMID- 8224777 TI - A C-terminal fragment of the hormone stanniocalcin is bioactive in eels. AB - An in vivo eel bioassay based on 45Ca uptake from the ambient medium was used to compare the activity of native trout and American eel stanniocalcin (STC) with that of a synthetic C-terminal fragment (AA 202-231; peptide W) of Australian eel STC. Qualitatively, the action of peptide W resembled that of purified native rainbow trout (Oncorhynchus mykiss) STC as well as fresh extracts of American eel Stannius corpuscles in that they all inhibited whole animal Ca2+ influx. This inhibition was observed in both intact and stanniectomized eels regardless of whether the eels were exhibiting high or low initial calcium uptake rates. These observations suggest that peptide W activates eel gill STC receptors. Further, as peptide W was previously reported to stimulate Ca2+ influx in intact rainbow trout fry, the present data provides evidence for a difference in the way this fragment functions in various species. PMID- 8224778 TI - 1,25(OH)2 vitamin D3 increases ionized plasma calcium concentrations in the immature Atlantic cod Gadus morhua. AB - Intraperitoneal injections of 1,25(OH)2 vitamin D3 (1,25(OH)2D3) produced hypercalcemia in the marine teleost the Atlantic cod (Gadus morhua) with increased plasma concentrations of ionized calcium (CaI) while the total plasma calcium concentrations (CaT) were unaffected. A single injection of 10 micrograms kg bw-1 of 1,25(OH)2D3 increased CaI concentrations from 1.74 +/- 0.03 mM to 1.8 +/- 0.01 mM after 24 hr in Experiment 1 and from 1.67 +/- 0.03 mM to 1.82 +/- 0.06 mM after 72 hr in Experiment 2. This hypercalcemic effect was sustained by daily injections for 5 but not 7 days. Daily injections of a lower dose of 1,25(OH)2D3, 1 microgram kg bw-1, caused hypercalcemia after 5 days (CaI increased from 1.68 +/- 0.01 mM to 1.76 +/- 0.02 mM). Plasma calcium concentrations were not affected by any of the other seco-steroids (vitamin D3, (25(OH)), vitamin D3 (25(OH)D3), or 24,25(OH)2 vitamin D3 (24,25(OH)2D3), except for a decrease in CaT concentrations after 7 daily in injections of 25(OH)D3 (2 micrograms kg bw-1). Plasma phosphate concentrations were not changed by any of the seco-steroids. It is concluded that 1,25(OH)2D3 is hypercalcemic in the marine Atlantic cod. PMID- 8224779 TI - Study of the complex formation between amine local anesthetics and uncouplers of oxidative phosphorylation carbonyl cyanide phenylhydrazones. AB - Spectroscopic evidence is presented which indicates that the anionic uncoupler carbonyl cyanide-4-nitro-2-chloro-phenylhydrazone and the amine local anesthetics form a complex in aqueous solution. The complex formation studies were carried out for several pharmacologically important tertiary amines and some primary amines. Their relative potencies to form a complex with uncoupler have followed the order: procaine < trimecaine < tetracaine < dibucaine < dodecylamine < dicyclohexylamine < hexadecylamine. As to the more lipophilic nature of the complex the emphasized penetration into octanol and reinforced retention into mitochondria was observed. The higher ability of the complex to colapse the mitochondrial membrane potential confirms this fact. The effective concentration of amine local anesthetics to form a complex was correlated with their physicochemical properties namely lipophilicity and acidobasicity. The highest effectivities for complex formation is shown by the most lipophilic and the most ionized molecules of amines. Present results point to the importance of considering the role of amine anesthetic-uncoupler complex in interpreting physiological or ion transport data in which these substances have been used together. PMID- 8224780 TI - Voltage dependent ionic currents in frog cultured skeletal myocytes. AB - The voltage dependent ionic currents in cultured embryonic skeletal myocytes at stages of development ranging from 1 to 6 day were studied using the whole-cell patch clamp technique. Sodium (INa) and calcium (ICa) inward and potassium (IK) outward currents were observed at all stages. INa did not differ from that described in adult frog striated muscle fibres. Slow ICa was mediated by current through dihydropyridine sensitive Ca channels and it did not differ in its kinetics from corresponding slow ICa in frog adult twitch muscle fibres. In about 10% of cells examined for ICa, this current was significantly slower and similar to ICa described in frog tonic muscle fibres. In some cases two slow calcium currents with distinguishable kinetics were recorded in the same myocytes. Fast dihydropyridine-insensitive noninactivating ICa could also be observed. At least 6 types of IK were registered, with approximate time-to-peak (at test pulse of 10 mV) 5, 12, 20, 30, 50 ms (fast IK) and more than 7 s (slow IK). Three of them (5, 20 and 30 ms) predominated in 3-day cultures and disappeared in 6-day-old cultures. IK in myocytes did not correspond fully in the kinetics to IK reported in adult frog skeletal muscles. Channels associated with transient fast and noninactivating slow IK were shown to be highly sensitive to low temperature (+5 degrees C). PMID- 8224781 TI - The interaction of amphotericin B with cell membrane of rat thymocytes. AB - Amphotericin B (AB) at micromolar concentrations increases cell membrane permeability and induced swelling of rat thymus lymphocytes. Potassium efflux is a precondition for AB to induce swelling of the cells. The rate constants for potassium loss and volume changes were proportional to the 1.24th and the 2nd power of the antibiotic concentration respectively. The reflection coefficients for nonelectrolytes with different hydrodynamic radii were determined, and the equivalent radius of the amphotericin pore in the thymocyte cell membrane was estimated to be 4.1 +/- 0.3 A at polyene concentrations varying between 2.5 mumol/l and 80 mumol/l. It is suggested that channel formation by AB in cell membranes is actually able to modulate immune responses. PMID- 8224782 TI - Effect of cholesterol, diacylglycerol and phosphatidylethanolamine on PEG 6000 induced lipid mixing and surface dielectric constant of phosphatidylcholine vesicle. AB - Using the NBD/Rh fluorescence assay the PEG 6000 induced phospholipid mixing of phosphatidylcholine liposomes containing cholesterol, diacylglycerol or phosphatidylethanolamine was measured. All 3 components shifted the PEG 6000 concentration necessary to induce 50% of maximal phospholipid mixing to lower concentrations. After the addition of PEG cholesterol containing PC liposomes exhibit different values of the surface dielectric constant as measured by the stokes shift of the fluorophore dansyl-PE compared to pure PC, whereas in DAG- and PE-containing PC liposomes no differences were observed. It is concluded that the incorporation of cholesterol leads to a different surface dielectric constant after PEG addition. The changed surface dielectric properties are a prerequest for the onset of fusion, as shown by Ohki and Arnold (1990). The incorporation of DAG and PE into PC membranes leads to structural instabilities as proposed by Siegel et al. (1989). This additional structurally unstable region created by molecules like PE or DAG may shift the onset of fusion to lower PEG concentration. PMID- 8224783 TI - Viscoelasticity of BLM from choline plasmalogen, alkylacyl- and diacyl glycerophosphocholines. AB - Mechanical characteristics of bilayer lipid membranes (BLM) composed of alkenylacyl-, alkylacyl-, and diacyl-glycerophosphocholines were studied by measuring modulus of elasticity in the direction normal to BLM plane, E perpendicular, and coefficient of dynamic viscosity eta. Alkenylacyl glycerophosphocholine (choline plasmalogen) BLM typically show larger values of E perpendicular and eta as compared to their analogs, suggesting a tighter packing of their hydrophobic regions. Increasing cholesterol concentrations are associated with monotonically increasing values of parameters E perpendicular and eta of plasmalogen BLM, whereas a nonmonotonic dependence of these parameters with a maximum around c = 66 mol% cholesterol is typical for the diacyl analog POPC. This may be due to the formation of cholesterol clusters at c > 66 mol%. PMID- 8224784 TI - Clustering of cholesterol in DMPC bilayers as indicated by membrane mechanical properties. AB - Mechanical characteristics of bilayer lipid membranes (BLM) composed of dimyristoylphosphatidylcholine (DMPC) and cholesterol in the gel and liquid crystalline state were studied by measuring the modulus of elasticity in direction perpendicular to the BLM plane, E perpendicular. The value of E perpendicular varied nonmonotonically with the cholesterol concentration, with a maximum around c = 50 mol% cholesterol. E perpendicular of BLM in gel state was about 2 times higher than that measured for the liquid crystalline state but the shape of E perpendicular (c) curves was similar for both states of the membrane. This may be due to the formation of cholesterol clusters at c > 50 mol% in both phase states of BLM. PMID- 8224785 TI - Blocking of potassium channels in Ranvier nodes by 4,5,6,7-substituted benzofurans and its significance on demyelinating diseases. PMID- 8224786 TI - Thenoyltrifluoracetone induces lipid peroxidation in rat liver mitochondria. AB - Addition of thenoyltrifluoracetone to mitochondria oxidizing NAD-dependent substrates induced lipid peroxidation and uncoupling of oxidative phosphorylation. Butylated hydroxytoluene, an antioxidant, prevented accumulation of lipid peroxidation products but failed to prevent deenergization of the mitochondria. PMID- 8224787 TI - Population genetics of a transformable bacterium: the influence of horizontal genetic exchange on the biology of Neisseria meningitidis. AB - Information of the biochemistry and genetics of bacterial species, usually obtained by the study of single isolates, is enhanced by studies of populations of bacteria. Recent advances in molecular technology, particularly polymerase chain reaction-based nucleotide sequence analysis, provide powerful tools for the study of population genetics. Data obtained by such techniques indicate that, while some bacterial species have a clonal population structure, others are non clonal or panmictic. Clonal populations are a consequence of asexual reproduction by binary fission; panmictic population structures result from 'horizontal' exchange of genetic material between clones. A consequence of horizontal genetic exchange is mosaic gene structures, recognisable by comparisons of nucleotide sequences. In transformable bacteria, for example the human pathogen Neisseria meningitidis, several different genes, including the gene encoding the class 1 outer membrane protein, a major surface antigen, are mosaics. This genetic process has implications both for vaccine design and in the interpretation of epidemiological data. PMID- 8224788 TI - Correlation between umuC induction and Salmonella mutagenicity assay for quinolone antimicrobial agents. AB - Quinolone antimicrobial agents induce the SOS response in bacteria, including the umuDC genes necessary for error-prone repair. Consequently these drugs may be mutagenic in bacteria with a functional SOS response. Differential killing tests with Escherichia coli WP2 (trp) and its repair-deficient derivative CM871 (trp lexA recA uvrA) indicated that a functional DNA repair system was protective against the action of quinolones, implying that quinolones are causing some form of DNA damage (not necessarily directly) and are therefore genotoxic. Dose dependent reversion from His- to His+ with quinolones was observed in the Ames test with Salmonella typhimurium TA102 (uvr+) but in no other Salmonella tester strains (all uvr-), suggesting that a functional excision repair system is essential for quinolone-induced bacterial mutagenesis. A significant correlation between SOS inducing potential (SOSIP) and mutagenic potential in the Ames test (r = 0.89) indicated that quinolone-induced mutagenic effects in bacteria are almost entirely due to SOS-processed DNA damage. PMID- 8224789 TI - Mapping of ben genes of Pseudomonas aeruginosa. AB - Four ben genes responsible for the conversion of benzoate to catechol in Pseudomonas aeruginosa PAO have been mapped to a 4.6 kb KpnI fragment, ben-1 and ben-4 were known to be separate genes but now ben-1508 has been found to be different from ben-2. The two genes were distinguished by Tn5 mutagenesis of a cosmid clone and deletion mapping. It is likely that the four genes mapped (ben 4, ben-2, ben-1508 and ben-1) correspond to the previously characterized benR (regulatory gene) and benABC (benzoate dioxygenase) respectively. PMID- 8224790 TI - Esterase from the oil-degrading Acinetobacter lwoffii RAG-1: sequence analysis and over-expression in Escherichia coli. AB - The est gene encoding an esterase from Acinetobacter lwoffii RAG-1 was cloned into E. coli under the control of the PL promoter of the phage lambda. The N terminal sequence of the first 20 amino acids of the heterologous expressed esterase corresponded to that obtained from the nucleotide sequence. Antibodies prepared against the over-expressed recombinant esterase in E. coli were used to locate the enzyme primarily in the membrane fractions of A. lwoffii RAG-1. Comparison with homologous proteins from both eukaryotic and prokaryotic organisms suggest that the RAG-1 esterase exhibits sequence motifs characteristic of both serine proteases and of lipases. PMID- 8224791 TI - Secretion of the Hormoconis resinae glucoamylase P enzyme from Trichoderma reesei directed by the natural and the cbh1 gene secretion signal. AB - Secretion of the Hormoconis resinae glucoamylase P (GAMP) enzyme from Trichoderma reesei using either the natural N-terminal extension of the premature glucoamylase P or the cellobiohydrolase I (CBHI) signal peptide was examined. The expression conditions for the heterologous glucoamylase P (gamP) gene in T. reesei were standardized by targeting one copy of a plasmid fragment, containing the gamP gene, to the cbh1 locus of the host. The results showed that the transient N-terminal extension of the premature GAMP acts as an efficient secretion signal in T. reesei and leads to a higher yield of extracellular glucoamylase activity than does the signal peptide of CBHI. PMID- 8224792 TI - Instability of waves formed by motile bacteria. AB - Many motile chemotactic bacteria (for instance, Escherichia coli) inoculated at some point in a semisolid nutrient medium can form circular expanding population waves. The formation of these motile structures is due to chemotaxis. The circular waves originate from an expanding bacterial lawn (a parent population). The regular shape of these waves results from the isotropic distribution of freely diffusible nutrient molecules which are also attractants. In this paper we show that the regular shape of the bacterial population waves can be spontaneously disturbed. As this takes place arc-shaped population waves ('bursts') are formed. It was found that initially the mean length of the cells forming the bursts was greater than that of the parent cell population. But then it decreased resulting in a value characteristic of the parent population. PMID- 8224793 TI - Pathological activities of Yersinia ruckeri, the enteric redmouth (ERM) bacterium. AB - The adherence and invasive capacities as well as the pathobiological activities exhibited by Yersinia ruckeri were examined. Although adhesive ability was dependent on the cell-line employed, all the strains showed moderate adhesion and invasiveness in the salmon cell-line CHSE-214. With regard to the extracellular products (ECP) all of them were strongly toxic for fish with LD50 ranging from 2 to 9.12 micrograms protein per g fish. In addition, all the ECP samples showed caseinase, gelatinase, amylase, lipase and phospholipase activities, hydrolysed esculin and displayed hemolytic activities for trout, salmon, sheep and human erythrocytes. Heat treatment (100 degrees C for 10 min) caused the loss of all these biological activities except the hydrolysis of gelatin. On the other hand, SDS-PAGE analysis of the LPS and protein components of the ECP revealed variations among strains depending on the serotype. The lack of lethal effects of the LPS present in the ECP was also demonstrated. PMID- 8224794 TI - Infectivity of a glucan synthesis-defective mutant of Streptococcus gordonii (Challis) in a rat endocarditis model. AB - Streptococcus gordonii, a member of the human indigenous oral microflora, colonizes smooth tooth surfaces and contributes to dental plaque formation. Although it is not recognized as being a cariogenic pathogen, it may cause endocarditis following invasion of the bloodstream. Using allelic exchange mutagenesis, we have constructed a mutant of S. gordonii (Challis) which is defective in its single functional glucosyltransferase gene and, hence, is unable to synthesize glucan exopolymers from sucrose. When examined in a rat endocarditis model, the sucrose-grown mutant did not differ significantly from S. gordonii wild-type, suggesting that glucan polymers did not contribute to infectivity. This result was in striking contrast to that previously observed with a polymer-defective S. mutans mutant. PMID- 8224795 TI - Raffinose-induced mutanase production from Trichoderma harzianum. AB - The enzyme alpha(1-->3),3-glucanohydrolase (referred to as mutanase) from the filamentous fungus Trichoderma harzianum OMZ 779 is capable of degrading the water-insoluble glucan in dental plaque. Previously, it was necessary to produce the glucan (referred to as mutan) in vitro for use as the sole carbon source and inducer of mutanase synthesis in fungal cultures. We report here that raffinose also induces the production of mutanase. The metabolism of raffinose differed from that of other sugars in metabolic end products and secreted protein profile. In addition to mutanase, we observed an approximately 15,000 M(r) protein that was also regulated by carbon source and by illumination conditions. PMID- 8224796 TI - Conjugal transfer of the determinants for bacteriocin (lacticin 481) production and immunity in Lactococcus lactis subsp. lactis CNRZ 481. AB - The lacticin 481-producer (Lct+), L. lactis subsp. lactis (L. lactis) CNRZ 481 harbours 5 plasmids of 6.5, 7.5, 20, 37 and 69 kb. Novobiocin treatment of L. lactis 481 led to the appearance of lacticin 481 deficient variants which had all lost the 69 kb plasmid. Conjugal transfer of the lacticin 481 structural gene (lct) into the plasmid free strain L. lactis IL1441 yielded Lct+ transconjugants at a 10(-4) frequency, which carried a plasmid with an apparent size of 120-130 kb. Southern hybridization analyses showed that the lct gene was located on the 69 kb plasmid in L. lactis 481 and on the 120-130 kb plasmid in the transconjugants. The lct gene was in higher copy number in transconjugants than in the parental strain resulting in two-fold higher lacticin 481 production in the former strain. PMID- 8224797 TI - Detection of molecular variation in the insect pathogenic fungus Metarhizium using RAPD-PCR. AB - DNA polymorphism among isolates of the insect pathogenic fungus Metarhizium anisopliae and M. flavoviride was investigated by RAPD-PCR. DNA fragments of between 0.3 and 2.7 kb were obtained using eight 10-mer PCR primers of arbitrary nucleotide sequence, and each isolate differed in the size and number of RAPD products, indicating considerable polymorphism. Isolate-specific RAPD fingerprints were used to calculate relative genetic similarity; this differentiated isolates into two major groups, separating nine of the ten isolates of M. anisopliae from the two of M. flavoviride. However, an Australian M. anisopliae isolated from an Orthopteran host exhibited a higher degree of genetic similarity to the M. flavoviride group. M. anisopliae isolates were further segregated into three subgroups which were loosely related to their geographical origins, although considerable polymorphism was observed within these groups. There was no apparent association between genotype and original insect host. PMID- 8224798 TI - The deficiency of sterol biosynthesis in Saccharomyces cerevisiae affects the synthesis of glycosyl derivatives of dolichyl phosphates. AB - Mutants deficient in sterol (thermosensitive ergosterol auxotrophs) erg 8, 9, 12 and heme synthesis hem 1, 12 were screened for the level of free dolichol and dolichyl phosphate synthesized in the mevalonate pathway as well as for the activity of dolichyl phosphate-dependent glycosyl transferases. The amount of DolP synthesized via CTP-dependent phosphorylation was the same in mutants and parental strains. However, mannosylation and glucosylation of endogenous dolichyl phosphates in ergosterol mutants was about four times lower compared to parental strains, while the same reactions carried out with exogenous Dol24P reached 80% of the level observed in parental strains indicating that activities of DolPMan and DolPGlc synthases are not the rate-limiting factors. It is postulated that the de novo synthesis of DolP is impaired in the ergosterol mutants. Moreover, a block in the ergosterol branch of the metabolic pathway (erg 9) causes an increase in the de novo synthesis of dolichyl phosphate. PMID- 8224799 TI - Anti-Candida activity of four antifungal benzothiazoles. AB - Anti-Candida activity of 6-amino-2-n-pentylthiobenzothiazole (I), benzylester of (6-amino-2-benzothiazolylthio)acetic acid (II) and of 3-butylthio-(1,2,4 triazolo)-2,3-benzothiazole (III) was followed and compared to that of 2 mercaptobenzothiazole (IV). I and II exhibited good activity against the C. albicans yeast form, similar to IV. They were inhibitorily active against other Candida strains, IC50 values being of the order of 10(-5) M, which means better activity than IV. Compound I also exhibited inhibitory activity on germ-tube formation and mycelial growth in the C. albicans strains, while II, III and IV were not active in these tests. III was the least active form of the compounds tested, IC50 values being of the order of 10(-4) M. All the compounds tested were highly active on a nystatin-resistant C. albicans mutant, with IC50s of the order of 10(-6) M-10(-5) M. PMID- 8224800 TI - Cloning, nucleotide sequence and expression of a beta-lactamase gene from Streptomyces lavendulae. AB - A hybridized DNA fragment was cloned as a 7.6-kb fragment from Streptomyces lavendulae KCCS0263 using a 1.9-kb SacI-XbaI DNA fragment from S. cellulosae as a probe. The latter fragment encoded a beta-lactamase which can bind blue dextran. The hybridized region was reduced to a 2.8-kb KpnI-BclI fragment and the nucleotide sequence was determined. The nucleotide sequence indicated one open reading frame whose amino acid sequence is very similar to that of the beta lactamase from S. cellulosae. The gene produced beta-lactamase enzyme at a low but significant amount. PMID- 8224801 TI - Detection of Naegleria fowleri cysts in environmental samples by using a DNA probe. AB - In this study we tried to detect DNA Naegleria fowleri in artificially contaminated environmental samples, with or without sediments, containing 10(4) cysts of this pathogenic amoeba. We used two assays to extract DNA from samples: first, direct DNA extraction, which gave positive results only for water samples without sediment; second, DNA extraction after sample incubation on agar plates, which allowed us to remove amoeba growing out of the sediments, and which gave positive results for all samples, even those initially with sediments (5, 500 or 500 mg). Thus, this molecular identification appears as a powerful tool to investigate N. fowleri growth in environmental samples. PMID- 8224802 TI - Chromosome-based method for rapid computer simulation in human genetic linkage analysis. AB - Computer-based simulation has been an important method in human linkage analysis for a long time. Typically, such analyses have been performed by simulating a set of linked markers according to the intermarker recombination fractions, under the assumption of no genetic interference. A novel approach is proposed in which such simulations can be performed using chromosome-based methods, rather than traditional recombination fraction-based methods. We propose simulating pedigree data using a crossover formation (CF) process to generate the number of crossovers and their locations in Morgans along the entire length of a chromosome. By this method, one can generate simulated multilocus data for any number of loci on a chromosome much more efficiently than with the currently available methods like those used in the SLINK or SIMLINK programs. Further, interference can be incorporated directly in this method, which is not possible with existing packages. PMID- 8224803 TI - Improved sib-pair linkage test for disease susceptibility loci. AB - An improved sib-pair test for linkage is introduced which is superior to the previously proposed tests. The test is derived from the standard chi-squared goodness of fit statistic by restricting the alternative hypothesis to the genetically possible. Critical values are given and exact power comparisons are made with the previously proposed tests. The new test is shown to be more powerful for finite samples as well as being asymptotically uniformly most powerful. PMID- 8224804 TI - Familial clustering of colon, breast, uterine, and ovarian cancers as assessed by family history. AB - The aggregation of colon, endometrial, ovarian, and possibly breast cancers in families has been described as a "cancer family syndrome" (now called Lynch syndrome II). To determine if the familial clustering of these malignancies was more common in women with cancer than without, we analyzed data from the Iowa Women's Health Study (IWHS), a population-based sample of 41,837 women aged 55-69 years. Self-reported information was collected on history of colon, uterine, ovarian, and breast cancers in female first-degree relatives. A family history of cancer of the breast (odds ratio [OR] = 1.4), colon (OR = 1.3), and uterus (OR = 1.3), but not ovary (OR = 1.2), was significantly more common among women with a personal history of any of these four cancers (all P < 0.05); the pattern of the ORs suggested strongly that the clustering tended to be site-specific. Age adjusted relative risks (RR) of incident colon cancer over 5 years of follow-up (N = 237) were calculated with regard to family history. Colon cancer incidence was increased among women with a family history of breast (RR = 1.3), uterine (RR = 1.4), colon (RR = 1.5), and ovarian (RR = 1.3) cancers, although none of the risk estimates achieved statistical significance. RR was, however, significantly related to the number of different cancer sites reported among family members (Ptrend = 0.008). These data on a representative sample of postmenopausal women suggest that family histories of colon, breast, uterine, and ovarian cancers are associated with an increased risk of cancer at the same site, but provide little support for the hypothesis that Lynch syndrome II is a non-random occurrence. PMID- 8224805 TI - Reporting bilaterality status in first-degree relatives with breast cancer: a validity study. AB - The objective of this study was to validate reports on the bilaterality status of breast cancer in first-degree relatives of women with a strong family history of the disease; i.e., women with 1) two first-degree relatives who have, or have had, breast cancer; 2) one first- and one second-degree affected relative; or 3) one first-degree relative with diagnosis of breast cancer before the age of 50 years and/or bilateral breast cancer. We were able to obtain hospital records for 94 affected relatives of 83 patients who agreed to participate in the study. The accounts of these women were compared to the bilaterality status indicated in the hospital records of the affected relatives. Inconsistencies that might have been attributed to incomplete medical records were resolved through personal interviews with the participants, and when indicated, with other family members or the physician of the affected relative. Overall, 89.4% (84/94) of the reports validated in this manner were correct. Participants who reported unilateral breast cancer in a first-degree relative were correct 94.4% (68/72) of the time. Similarly, 94.0% (47/50) of the accounts concerning affected living relatives were accurate, regardless of whether the participant had indicated unilateral or bilateral disease. However, participants who reported bilateral breast cancer in a deceased relative were accurate only 61.5% (8/13) of the time. Incorrect reports were associated with misunderstanding of medical terminology, especially if the participant was young at the time of the diagnosis of her relative. PMID- 8224806 TI - Genotype at a major locus with large effects on apolipoprotein B levels predicts familial combined hyperlipidemia. AB - A sample enriched for familial combined hyperlipidemia (FCHL) was examined for evidence of an association between genotype at an apolipoprotein B (apoB) elevating locus defined by complex segregation analysis and FCHL. Complex segregation analysis detected a locus with a large effect on plasma apoB levels and was used to compute the most probable genotype of family members. None of the 35 normolipidemic adults carried a copy of the allele associated with elevated apoB levels, yet 58% of the 109 adults with FCHL carried 1 (29%) or 2 (28%) copies. Two of 28 (7%) normal children had 1 copy of this allele and none had 2 copies, while 88 of 182 (48%) children with FCHL had 1 (26%) or 2 (22%) copies. Further, 41 of 48 (85%) individuals classified as having hyperapobetalipoproteinemia did not carry a copy of this "elevated apoB" allele. Therefore, the presence of the allele associated with elevation of apoB level is highly predictive of FCHL and this association cannot be explained solely by the presence of elevated apoB levels in FCHL, suggesting that the locus controlling apoB levels may play an etiologic role in FCHL. PMID- 8224807 TI - Homozygous parent affected sib pair method for detecting disease predisposing variants: application to insulin dependent diabetes mellitus. AB - For complex genetic diseases involving incomplete penetrance, genetic heterogeneity, and multiple disease genes, it is often difficult to determine the molecular variant(s) responsible for the disease pathogenesis. Linkage and association studies may help identify genetic regions and molecular variants suspected of being directly responsible for disease predisposition or protection, but, especially for complex diseases, they are less useful for determining when a predisposing molecular variant has been identified. In this paper, we expand upon the simple concept that if a genetic factor predisposing to disease has been fully identified, then a parent homozygous for this factor should transmit either of his/her copies at random to any affected children. Closely linked markers are used to determine identity by descent values in affected sib pairs from a parent homozygous for a putative disease predisposing factor. The expected deviation of haplotype sharing from 50%, when not all haplotypes carrying this factor are in fact equally predisposing, has been algebraically determined for a single locus general disease model. Equations to determine expected sharing for multiple disease alleles or multiple disease locus models have been formulated. The recessive case is in practice limiting and therefore can be used to estimate the maximum proportion of putative susceptibility haplotypes which are in fact predisposing to disease when the mode of inheritance of a disease is unknown. This method has been applied to 27 DR3/DR3 parents and 50 DR4/DR4 parents who have at least 2 children affected with insulin dependent diabetes mellitus (IDDM). The transmission of both DR3 and DR4 haplotypes is statistically different from 50% (P < 0.05 and P < 0.001, respectively). An upper estimate for the proportion of DR3 haplotypes associated with a high IDDM susceptibility is 49%, and for DR4 haplotypes 38%. Our results show that the joint presence of non Asp at DQ beta position 57 and Arg at DQ alpha position 52, which has been proposed as a strong IDDM predisposing factor, is insufficient to explain the HLA component of IDDM predisposition. PMID- 8224808 TI - AGFAP method: applicability under different ascertainment schemes and a parental contributions test. AB - The antigen/allele genotype frequencies among patients (AGFAP) method has been powerful in discriminating between modes of inheritance, and detecting heterogeneity effects, for a number of diseases associated with the HLA system. The method is not dependent on the high level of polymorphism seen in the HLA system, but does require a marker allele association with disease. With recent rapid advances in mapping of the human genome, the method is increasingly relevant in all disease studies. Extension of the AGFAP method to ascertainment schemes other than random sampling of patients is presented here. The method is shown to be robust for distinguishing between incompletely penetrant recessive vs. additive or dominant models if affected children are obtained from nuclear families selected on the basis of at least two affected members: two affected sibs, or an affected parent and affected child. The method can lead to false conclusions for data from families ascertained for at least one affected parent and two affected children. A new test, termed the parental contributions test, applicable in families selected for the presence of an affected parent, and one or more affected children, is presented. The test, based on the expected symmetry (recessive) vs. asymmetry (additive and dominant) of parental marker allele contributions to an affected offspring in these pedigrees, is powerful in distinguishing between these modes of inheritance when there is a marker allele association with disease. Sporadic cases of disease are shown to cause deviations from AGFAP expectations for the recessive model, but not for the additive model. These results will aid in study of the genetics, and hence molecular basis, of complex diseases. PMID- 8224809 TI - Epistatic modeling in rheumatoid arthritis: an application of the Risch theory. AB - Rheumatoid arthritis (RA) is a disease of unknown etiology but with a presumed complex pattern of inheritance. Risch [Am J Hum Genet 46:222-228, 1990] has shown that the recurrence risk ratio, lambda R, (which is defined as the risk to type R relatives vs. the population prevalence) can be used to evaluate patterns of inheritance in genetically complex diseases. We have used the Risch theory to examine some multiple locus models of inheritance in RA. Recurrence risk ratios in MZ twins and in 1st, 2nd, and 3rd degree relatives are summarized from the literature. The limited data available supports at least a two-locus model of inheritance for RA (assuming that one locus is HLA). Better estimates of the recurrence risk ratios in RA families are required so that the Risch theory can be pursued further. PMID- 8224810 TI - Scope and strategies of genetic epidemiology: analysis of articles published in Genetic Epidemiology, 1984-1991. AB - Genetic epidemiology is a relatively new discipline that seeks to unravel the role of genetic factors and their interactions with environmental factors in the etiology of diseases, using population and family study approaches. To characterize the overall direction and emphasis of research strategies used in this field, we reviewed original research articles published in the journal Genetic Epidemiology since its inception in 1984 until the end of 1991. Of 259 published original articles, 92 (35%) focused primarily on methodologic/statistical developments, most commonly in the area of linkage analysis/gene mapping, and 167 (65%) articles were applied or data-derived. Only 42 articles (16%) were population studies, and 217 (84%) were family studies. Most family studies dealt with genetic analysis of pedigree data using segregation and linkage analyses. Of the 137 applied family studies, 73 (53%) were drawn from well-defined populations, and only 40 (29%) considered specific environmental factors in their analyses. These findings clearly indicate a rapid growth in the methodologic and statistical aspects of genetic epidemiology, and in the emphasis on family-based studies and genetic analysis methods. Further developments in genetic epidemiology will require greater integration of epidemiologic approaches of study design and analyses into population and family studies of disease etiology. PMID- 8224811 TI - Francis Galton: count and measure, measure and count. PMID- 8224812 TI - Genetic and cytogenetic analysis of the 43A-E region containing the segment polarity gene costa and the cellular polarity genes prickle and spiny-legs in Drosophila melanogaster. AB - A cytogenetic analysis of the 43A-E region of chromosome 2 in Drosophila melanogaster is presented. Within this interval 27 complementation groups have been identified by extensive F2 screens and ordered by deletion mapping. The region includes the cellular polarity genes prickle and spiny-legs, the segmentation genes costa and torso, the morphogenetic locus sine oculis and is bounded on its distal side by the eye-color gene cinnabar. In addition 19 novel lethal complementation groups and two semi-lethal complementation groups with morphogenetic escaper phenotypes are described. PMID- 8224813 TI - Mutations in the protein phosphatase 1 gene at 87B can differentially affect suppression of position-effect variegation and mitosis in Drosophila melanogaster. AB - The suppressor of position effect variegation (PEV) locus Su-var(3)6 maps to 87B5 10. The breakpoints of deficiencies that define this interval have been placed on a 250-kb molecular map of the region. The locus is allelic to the ck19 complementation group previously shown to encode a type 1 serine-threonine protein phosphatase (PP1) catalytic subunit. When introduced into flies by P element-mediated transformation, a 5.8-kb genomic fragment carrying this gene overcomes the suppressor phenotype of Su-var(3)6(01) and recessive lethality of all mutations of the locus. Four of the mutant alleles at the locus show a broad correlation between high levels of suppression of PEV, a high frequency of aberrant mitosis and low PP1 activity in larval extracts. However, some alleles with low PP1 activity show weak suppression of PEV with a high frequency of abnormal mitosis, whereas others show strong suppression of PEV with normal mitosis. The basis for these discussed. PMID- 8224814 TI - A combined classical genetic and high resolution two-dimensional electrophoretic approach to the assessment of the number of genes affecting hybrid male sterility in Drosophila simulans and Drosophila sechellia. AB - We have attempted to estimate the number of genes involved in postzygotic reproductive isolation between two closely related species, Drosophila simulans and Drosophila sechellia, by a novel approach that involves the use of high resolution two-dimensional gel electrophoresis (2DE) to examine testis proteins in parents, hybrids and fertile and sterile backcross progenies. The important results that have emerged from this study are as follows: (1) about 8% of about 1000 proteins examined showed divergence (presence/absence) between the two species; (2) by tracing individual proteins in parental, hybrid and backcross males, we were able to associate the divergent proteins with different chromosomes and found that most divergent proteins are associated with autosomes and very few with X chromosome, Y chromosome and cytoplasm; (3) when proteins showing both quantitative and qualitative differences between the two species were examined in F1 hybrid males, most (97.4%) proteins were expressed at levels between the two parents and no sign of large scale changes in spot density was observed. All the proteins observed in the two parental species were present in F1 hybrid males except two species-specific proteins that may be encoded (or regulated) by sex chromosomes; (4) when different fertile and sterile backcross male testes were compared, a few D. sechellia-specific proteins were identified to be consistently associated with male sterility. These results along with the observation that a large proportion (23.6%) of first generation backcross males were fertile show that hybrid male sterility between D. simulans and D. sechellia involves a relatively small number of genes. Role of large scale genetic changes due to general genome incompatibility is not supported. The results also suggest that the large effect of X chromosome on hybrid male sterility is not due to higher divergence of X chromosome than autosomes. PMID- 8224815 TI - Maternal repression of the P element promoter in the germline of Drosophila melanogaster: a model for the P cytotype. AB - The transposition of P elements in Drosophila melanogaster is regulated by products encoded by the P elements themselves. The P cytotype, which represses transposition and associated phenomena, exhibits both a maternal effect and maternal inheritance. The genetic and molecular mechanisms of this regulation are complex and not yet fully understood. In a previous study, using P-lacZ fusion genes, we have shown that P element regulatory products were able to inhibit the activity of the P promoter in somatic tissues. However, the repression observed did not exhibit the maternal effect characteristic of the P cytotype. With a similar approach, we have assayed in vivo the effect of P element regulatory products in the germline. We show that the P cytotype is able to repress the P promoter in the germline as well as in the soma. Furthermore, this repression exhibits a maternal effect restricted to the germline. On the basis of these new observations, we propose a model for the mechanism of P cytotype repression and its maternal inheritance. PMID- 8224816 TI - A mutation affecting the lactate dehydrogenase locus Ldh-1 in the mouse. II. Mechanism of the LDH-A deficiency associated with hemolytic anemia. AB - A procarbazine hydrochloride-induced mutation at the Ldh-1 structural locus encoding the A subunit of lactate dehydrogenase (LDH) was used to study the molecular and metabolic basis of severe hemolytic anemia due to LDH-A deficiency in the mouse. The mutant allele designated Ldh-1a-m1Neu codes for an enzyme that as homotetramer differs from the wild-type enzyme by a marked instability, acidic shift of the pH profile, increased Km for pyruvate and altered inhibition by high concentrations of this substrate. Except for the latter, all these altered properties of the mutant protein contribute to the diminished LDH activity in heterozygous and homozygous mutant individuals. Impaired energy metabolism of erythrocytes indicated by a relatively low ATP concentration is suggested to result in cell death at the end of the reticulocyte stage leading to the expression of hemolytic anemia with extreme reticulocytosis and hyperbilirubinemia. Despite the severe anemia, affected homozygous mutants exhibit approximately normal body weight and do not show noticeable impairment of viability or fertility. To date no such condition is observed in man. This discrepancy is likely due to the fact that in human erythrocytes both LDH-A and LDH-B subunits are expressed such that homozygotes for a LDH-A or LDH-B deficiency would not result in a comparably extreme LDH activity deficiency. PMID- 8224817 TI - Molecular and genetic analysis of the SNF7 gene in Saccharomyces cerevisiae. AB - Mutations in the SNF7 gene of Saccharomyces cerevisiae prevent full derepression of the SUC2 (invertase) gene in response to glucose limitation. We report the molecular cloning of the SNF7 gene by complementation. Sequence analysis predicts that the gene product is a 27-kDa acidic protein. Disruption of the chromosomal locus causes a fewfold decrease in invertase derepression, a growth defect on raffinose, temperature-sensitive growth on glucose, and a sporulation defect in homozygous diploids. Genetic analysis of the interactions of the snf7 null mutation with ssn6 and spt6/ssn20 suppressor mutations distinguished SNF7 from the SNF2, SNF5 and SNF6 genes. The snf7 mutation also behaved differently from mutations in SNF1 and SNF4 in that snf7 ssn6 double mutants displayed a synthetic phenotype of severe temperature sensitivity for growth. We also mapped SNF7 to the right arm of chromosome XII near the centromere. PMID- 8224818 TI - Variance analysis of immunoglobulin alleles in natural populations of rabbit (Oryctolagus cuniculus): the extensive interallelic divergence at the b locus could be the outcome of overdominance-type selection. AB - Population genetic data are presented which should contribute to evaluation of the hypothesis that the extraordinary evolutionary patterns observed at the b locus of the rabbit immunoglobulin light chain constant region can be the outcome of overdominance-type selection. The analysis of allele correlations in natural populations revealed an excess of heterozygotes of about 10% at the b locus while heterozygote excess was not observed at loci determining the immunoglobulin heavy chain. Data from the published literature, where homozygote advantage was suggested, were reevaluated and found in agreement with data here presented. Gene diversity was evenly distributed among populations and showed similarities with patterns reported for histocompatibility loci. Analysis of genotypic disequilibria revealed strong digenic associations between the leading alleles of heavy and light chain constant region loci in conjunction with trigenic disequilibria corresponding to a preferential association of b locus heterozygosity with the predominant allele of the heavy chain e locus. It is argued that this may indicate compensatory or nonadditive aspects of a putative heterozygosity enhancing mechanism, implying that effects at the light chain might be more pronounced in populations fixed for the heavy chain polymorphism. PMID- 8224820 TI - Interval mapping of multiple quantitative trait loci. AB - The interval mapping method is widely used for the mapping of quantitative trait loci (QTLs) in segregating generations derived from crosses between inbred lines. The efficiency of detecting and the accuracy of mapping multiple QTLs by using genetic markers are much increased by employing multiple QTL models instead of the single QTL models (and no QTL models) used in interval mapping. However, the computational work involved with multiple QTL models is considerable when the number of QTLs is large. In this paper it is proposed to combine multiple linear regression methods with conventional interval mapping. This is achieved by fitting one QTL at a time in a given interval and simultaneously using (part of) the markers as cofactors to eliminate the effects of additional QTLs. It is shown that the proposed method combines the easy computation of the single QTL interval mapping method with much of the efficiency and accuracy of multiple QTL models. PMID- 8224819 TI - Somatic variegation and germinal mutability reflect the position of transposable element Dissociation within the maize R gene. AB - The R gene regulates the timing and tissue-specificity of anthocyanin deposition during maize development. The Ac/Ds system of transposable elements was used to induce insertional mutants of the R-sc:124 allele during two cycles of mutagenesis. Of 43 unstable, spotted-aleurone mutants generated, 42 contain inserts of the Ds6 transposable element differing only in the position and orientation of the element. The remaining mutant, r-sc:m1, contained an insert of a Ds element of the approximate size of the Ds1 transposable element. The patterns of somatic variegation of these mutants, resulting from excision of Ds, define a spectrum of phenotypes ranging from sparse to dense variegation. The sparsely variegated mutants produce few germinal revertants but relatively many stable null derivative alleles; densely variegated mutants produce many germinal revertants and few stable null derivatives. Molecular analysis shows that the sparsely variegated alleles are caused by Ds6 insertions in protein coding regions of R-sc:124 whereas the densely variegated mutants result from insertions in introns or in flanking regions of the gene. The excision rate of Ds6 from R, estimated as the proportion of R genomic DNA restriction fragments lacking the element, was uniform regardless of position, orientation or whether the element was inserted in R-sc:124 or another R allele. The excision rate was greater, however, for the mutable alleles involving the Ds element from r-sc:m1. These data indicate that, although the excision rates are uniform for a given Ds element, the somatic and germinal mutability patterns of alleles associated with that element vary widely and depend primarily on the position of the transposable element within coding or noncoding regions of the gene. PMID- 8224821 TI - A model simulating the dynamics of plant mitochondrial genomes. AB - Molecular evolution of the plant mitochondrial genome involves rearrangements due to the presence of highly recombining repeated sequences. As a result, this genome is composed of a set of molecules of various sizes that generate each other through recombination. The model presented simulates the evolution of various frequencies of the different types of molecules over successive cell cycles. It considers the mitochondrial genome as a population of circular molecules evolving through recombination, replication and random segregation. The model parameters are the rates of recombination of each sequence, the frequency of each type of recombination, the replication rates of the circles and the total amount of mitochondrial DNA per cell. This model demonstrates that high recombination rates lead to rapid deletions of sequences in the absence of selection. The frequency of deletion is dependent on the simulated reproductive mechanism. The conditions leading to reversible or irreversible rearrangements were also investigated. PMID- 8224822 TI - Efficiency of generations for estimating marker-associated QTL effects by multiple regression. AB - Knowledge about the efficiency of generations for estimating marker-associated QTLs is needed for selection. The objective of this paper is to develop a theory to compare the efficiency of segregating generations and testcrosses from the cross of two inbred lines differing in value for a quantitative trait (P1 x P2) for estimating additive, dominance and heterotic effects of QTLs by stepwise regression. An equation that predicts the smallest gene effect in genetic standard deviation units that can be detected with 50% chance at a significance level as a function of the heritability (h2) and the recombination frequency (r) of markers was developed for the segregating generations and testcrosses. For estimating additive effects, the most efficient generation was the doubled haploid (DH) lines; the most inefficient was the North Carolina Design III (NCD III), followed by selfed backcrosses (SB); the selfed families from F2 individual plants (F2:3 lines) are inferior to the recombinant inbreds (RI) for low r, but are better than RI for high h2 and r. Dominance effects are less efficiently estimated than additive effects. The NCD III is better than the SB and the F2:3 lines for detecting dominance effects. The RI and DH do not estimate dominance effects. The differential heterotic QTL effects of lines P1 and P2 when crossed with tester T can be estimated by evaluating testcrosses of individual F2 plants (F2T), recombinant inbreds (RIT) and double-haploid lines (DHT). The DHT is superior to the other generations. The F2T is better than the RIT for r > or = 0.20, but inferior for r < or = 0.1 or low heritability. PMID- 8224823 TI - A quantitative test of the neutral theory using pooled allozyme data. AB - Neutral theory predicts a positive correlation between the amount of polymorphism within species and evolutionary rate. Previous tests of this prediction using both allozyme and DNA data have led to conflicting conclusions about the influence of selection and mutation drift. It is argued here that quantitative conclusions about the adequacy of neutral theory can be obtained by analyzing genetic data pooled from many sources. Using this approach, a large database containing information on allozyme variation in over 1500 species is used to examine the relationship between heterozygosity and genetic distance. The results provide support for the hypothesis that a major percentage of protein variation can be explained by variation in neutral mutation rate, and a minor percentage by strong selection. PMID- 8224824 TI - The HIR4-1 mutation defines a new class of histone regulatory genes in Saccharomyces cerevisiae. AB - Both activation and repression have been implicated in the cell cycle-regulated transcription of the histone HTA1-HTB1 locus in Saccharomyces cerevisiae. Transcriptional repressors have been identified through the isolation of recessive mutations in the HIR1, HIR2 and HIR3 genes. These three regulatory genes encode proteins that act at a negative site in the HTA1-HTB1 promoter, and their inactivation results in cell cycle-independent transcription. We report here on the characterization of a fourth HIR mutant. The HIR4-1 mutation is dominant, and the phenotypes that it confers suggest that the mutant gene encodes an altered transcriptional activator. The function of this activator is very specific: it uniquely regulates transcription of the HTA1-HTB1 locus, and it may antagonize repressors that act through the HTA1-HTB1 negative site. PMID- 8224825 TI - Loss of function of Saccharomyces cerevisiae kinesin-related CIN8 and KIP1 is suppressed by KAR3 motor domain mutations. AB - The kinesin-related products of the CIN8 and KIP1 genes of Saccharomyces cerevisiae redundantly perform an essential function in mitosis. The action of either gene-product is required for an outwardly directed force that acts upon the spindle poles. We have selected mutations that suppress the temperature sensitivity of a cin8-temperature-sensitive kip1-delta strain. The extragenic suppressors analyzed were all found to be alleles of the KAR3 gene. KAR3 encodes a distinct kinesin-related protein whose action antagonizes Cin8p/Kip1p function. All seven alleles analyzed were altered within the region of KAR3 that encodes the putative force-generating (or "motor") domain. These mutations also suppressed the inviability associated with the cin8-delta kip1-delta genotype, a property not shared by a deletion of KAR3. Other properties of the suppressing alleles revealed that they were not null for function. Six of the seven were unaffected for the essential karyogamy and meiosis properties of KAR3 and the seventh was dominant for the suppressing trait. Our findings suggest that despite an antagonistic relationship between Cin8p/Kip1p and Kar3p, aspects of their mitotic roles may be similar. PMID- 8224826 TI - A single amino acid difference is sufficient to elicit vegetative incompatibility in the fungus Podospora anserina. AB - Vegetative incompatibility is known to limit heterokaryosis in filamentous fungi. It results from genetic differences between incompatible strains at specific loci. The proteins encoded by the two incompatible alleles het-s and het-S of the fungus Podospora anserina differ from each other by 14 amino acids. Two approaches have been used to identify how many and which of these differences are necessary to elicit incompatibility. Twelve alleles of the het-s locus of wild type isolates of P. anserina and of the related species Podospora comata have been sequenced to determine the extent of the variability of genes controlling s and S specificities. Expression of hybrid het-s/het-S genes and site-specific mutagenesis revealed that the specificities of het-s and het-S are under the control of a limited number of amino acid differences. The results show that vegetative incompatibility between s and S strains can be attributed to a single amino acid difference in the proteins encoded by the het-s locus. PMID- 8224827 TI - Gene conversions and crossing over during homologous and homeologous ectopic recombination in Saccharomyces cerevisiae. AB - The pma1-105 mutation reduces the activity of the yeast plasma membrane H(+) ATPase and causes cells to be both low pH and ammonium ion sensitive and resistant to the antibiotic hygromycin B. Revertants that can grow at pH 3.0 and on ammonium-containing plates frequently arise by ectopic recombination between pma1-105 and PMA2, a diverged gene that shares 85% DNA sequence identity with PMA1. The gene conversion tracts of revertants of pma1-105 were determined by DNA sequencing the hybrid PMA1::PMA2 genes. Gene conversion tracts ranged from 18-774 bp. The boundaries of these replacements were short (3-26 bp) regions of sequences that were identical between PMA1 and PMA2. These boundaries were not located at the regions of greatest shared identity between the two PMA genes. Similar results were obtained among low pH-resistant revertants of another mutation, pma1-147. One gene conversion was obtained in which the resulting PMA1::PMA2 hybrid was low pH-resistant but still hygromycin B-resistant. This partially active gene differs from a wild-type revertant only by the presence of two PMA2-encoded amino acid substitutions. Thus, some regions of PMA2 are not fully interchangeable with PMA1. We have also compared the efficiency of recombination between pma1-105 and either homeologous PMA2 sequence or homologous PMA1 donor sequences inserted at the same location. PMA2 x pma1-105 recombination occurred at a rate approximately 75-fold less than PMA1 x pma1-105 events. The difference in homology between the interacting sequences did not affect the proportion of gene conversion events associated with a cross-over, as in both cases approximately 5% of the Pma+ recombinants had undergone reciprocal translocations between the two chromosomes carrying pma1-105 and the donor PMA sequences. Reciprocal translocations were identified by a simple and generally useful nutritional test. PMID- 8224828 TI - Three new classes of mutations in the Caenorhabditis elegans muscle gene sup-9. AB - We are studying five interacting genes involved in the regulation or coordination of muscle contraction in Caenorhabditis elegans. A distinctive "rubber-band" muscle-defective phenotype was previously shown to result from rare altered function mutations in either of two of these genes, unc-93 and sup-10. null mutations in sup-9, sup-10, sup-18 or unc-93 act as essentially recessive suppressors of these rubber-band mutations. In this work, we identify three new classes of sup-9 alleles: altered-function rubber-band, partial loss-of-function and dominant-suppressor. The existence of rubber-band mutations in sup-9, sup-10 and unc-93 and the suppression of these mutations by null mutations in any of these three genes suggest that these proteins are required at the same step in muscle contraction. Moreover, allele-specific interactions shown by the partial loss-of-function mutations indicate that the products of these interacting genes may physically contact each other in a multiple subunit protein complex. Finally, the phenotypes of double rubber-band mutant combinations suggest that the rubber band mutations affect a neurogenic rather than a myogenic input in excitation contraction coupling in muscle. PMID- 8224829 TI - P-lacW insertional mutagenesis on the second chromosome of Drosophila melanogaster: isolation of lethals with different overgrowth phenotypes. AB - A single P-element insertional mutagenesis experiment was carried out for the second chromosome of Drosophila melanogaster using the P-lacW transposon. Out of 15,475 insertions on the second chromosome, 2,308 lethal and 403 semilethal mutants (altogether 2,711) were recovered. After eliminating clusters, 72% of the mutants represent independent insertions. Some of the mutants with larval, prepupal or pupal lethal phases have a prolonged larval period and show gradual overgrowth of the imaginal discs, brain and/or the hematopoietic organs (lymph glands). In this paper, 16 overgrowth mutants are described. As revealed by in situ hybridization, none of the mutations corresponds to any of the previously known overgrowth mutations on the second chromosome. PMID- 8224830 TI - Type I repressors of P element mobility. AB - We describe here a family of P elements that we refer to as type I repressors. These elements are identified by their repressor functions and their lack of any deletion within the first two-thirds of the canonical P sequence. Elements belonging to this repressor class were isolated from P strains and were made in vitro. We found that type I repressor elements could strongly repress both a cytotype-dependent allele and P element mobility in somatic and germline tissues. These effects were very dependent on genomic position. Moreover, we observed that an element's ability to repress in one assay positively correlated with its ability to repress in either of the other two assays. The type I family of repressor elements includes both autonomous P elements and those lacking exon 3 of the P element. Fine structure deletion mapping showed that the minimal 3' boundary of a functional type I element lies between nucleotide position 1950 and 1956. None of 12 elements examined with more extreme deletions extending into exon 2 made repressor. We conclude that the type I repressors form a structurally distinct group that does not include more extensively deleted repressor elements such as the KP element described previously. PMID- 8224831 TI - RNA folding in Drosophila shows a distance effect for compensatory fitness interactions. AB - Phylogenetic-comparative analysis was used to construct a secondary structure of Adh precursor messenger RNA (pre-mRNA) in Drosophila. The analysis revealed that the rate of coevolution of base-pairing residues decreases with their physical distance. This result is in qualitative agreement with a model of compensatory fitness interactions which assumes that mutations are individually deleterious but become harmless (neutral) in appropriate combinations. This model predicts that coupled mutations can become fixed in a population under mutation pressure and random genetic drift, when the mutations are closely linked. However, the rate of joint fixation drops as distance between sites increases and recombination breaks up favorable combinations. RNA secondary structure was also used to interpret patterns of linkage disequilibrium at Adh. PMID- 8224832 TI - Protease activity in cockroach and basidiomycete allergen extracts. AB - Inherent proteolytic activity was estimated in cockroach and basidiomycete extracts by quantifying acid soluble peptides that were released by incubating extracts with 1% bovine serum albumin as measured by Lowry (Sigma). Reference proteases released 740 (Proteinase K, 0.1 U), 248 (Trypsin, 1.0 U), and 533 micrograms/ml (Pronase, 0.5 U) of soluble peptides. American whole body cockroach extract (0.1 mg dry weight) released 330 micrograms/ml of soluble peptides, representing 13 trypsin equivalent units (TEU)/mg. Extracts from spores of the mushroom Pleurotus ostreatus released 230 micrograms/ml (0.9 TEU/mg) and Pleurotus cap extract released 112 micrograms/ml (0.5 TEU/mg). Mycelium of Pleurotus and the mushroom Psilocybe cubensis and spores of Psilocybe and the puffball Calvatia cyathiformis showed negligible amounts of proteolytic activity. The protease inhibitor phenylmethylsulfonyl flouride reduced the proteolytic activity of American whole body cockroach extract by 80% (@1 mM) and the inhibitor ethylene diaminetetraacetic acid inhibited the proteolytic activity of Pleurotus spores by 95% (@1 mM). Loss of allergen activity as determined by RAST inhibition and immunoprinting correlated with protease activity. Thus, in the preparation and handling of allergen extracts, one should employ conditions that minimize proteolysis. PMID- 8224833 TI - ELISA screening tests for specific IgE and IgG antibody in employees exposed to trimellitic anhydride (TMA). AB - The objective of the study was to determine whether ELISA screening tests could be developed for specific IgE and IgG antibody in surveillance studies of employees exposed to trimellitic anhydride (TMA). Such tests would obviate the need for radiolabeling and disposal of radioactive wastes. The design of the study was a masked comparison of serologic results of ELISA screening tests with the criterion standard radioimmunoassay (RIA) currently used. The participants were 233 employees of a chemical plant that manufactures TMA. When the screening ELISA serologic results for IgG against TM-human serum albumin (TM-HSA) were compared with those of the RIA, the sensitivity was 92%, the specificity was 93%, the positive predictive value was 62%, and the negative predictive value was 99%. When the screening ELISA serologic results for IgE against TM-HSA were compared with those of the RIA, the sensitivity was 100%, the specificity was 91%, the positive predictive value was 31%, and the negative predictive value was 100%. We conclude that ELISA screening tests for antibody to TM-HSA have sufficient sensitivity, specificity, and predictive value to be useful in surveillance studies of employees exposed to TMA. PMID- 8224834 TI - Nasal hyperreactivity and its effect on early and late sequelae of nasal challenge with house-dust mite extract. AB - For a study on the relationship between nasal hyperreactivity to histamine and the nasal response to allergen, 14 rhinitis patients allergic to house-dust mites were challenged with histamine and 5 days later with a house-dust mite (HDM) extract. According to symptom scores, after allergen challenge two groups of patients were distinguished, i.e., isolated early and dual responders. The nasal response to histamine was significantly correlated with the amount of secretion (r = 0.71; p = 0.0039) and the number of sneezes (r = 0.78; p = 0.0016) induced by the HDM extract during the early reaction. The amount of allergen-induced secretion could be predicted from the response to histamine, skin reactivity to allergen, and blood eosinophils (multiple r = 0.90; p < 0.0001). Late-phase symptoms appearing between 3.5 and 9.5 hour after allergen challenge could be predicted from histamine responsiveness and skin reactivity (multiple r = 0.67; p = 0.004). Compared with early responders (LAR-) (n = 8), patients with early and late symptoms (LAR+) (n = 6) were characterized by a higher secretory responsiveness to histamine (p = 0.033), increased production of leukotrienes determined in nasal lavage fluid during the early response (p = 0.033), and elevated albumin levels occurring between 3.5 and 9.5 hours after challenge (p = 0.043). Late-phase symptoms were significantly correlated with albumin influx (r = 0.73; p = 0.001) and leukotrienes production (r = 0.60; p = 0.011) during the early reaction. In summary, nasal responsiveness to HDM extract was found to be closely associated with pre-existent nasal hyperreactivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224835 TI - Extended evaluation of diagnostic skin testing practices in Orange County, California. AB - In an effort to evaluate conformity with published guidelines for diagnostic allergy skin testing in their locality, a committee of members of the Orange County (California) Society of Allergy and Clinical Immunology surveyed their membership for the number, identity, and rates of sensitization of aeroallergens in a selected patient population. Test data were analyzed on patients who were judged appropriate to undergo immunotherapy. Complete test results of 271 patients were obtained from one third of local allergy specialists. A total of 115 different aeroallergen extracts were identified. By category, tree pollen allergens (32) were followed in declining order by weed pollens (28), molds (22), environmentals (18), and grass pollen extracts (15), for the total of 115. The average number of aeroallergens used by a practice in a comprehensive evaluation was 77 (range, 47-114). When only unequivocally positive reactions were considered (3+ equivalent or greater), Bermuda grass pollen extracts elicited the highest rate, 85%, and the environmental, rat dander, was the lowest at 1.8%. At least 20% of the patients reacted strongly on skin tests to 97% of the extracts. The disparity between the recommendations of a maximum of 30 aeroallergens for a comprehensive evaluation and actual practice procedures merits efforts at resolution. PMID- 8224836 TI - Fish, in and out of water; food, toxins, allergens. PMID- 8224837 TI - Proposed NF-kappa B/I kappa B family nomenclature. PMID- 8224838 TI - The I kappa B proteins: multifunctional regulators of Rel/NF-kappa B transcription factors. PMID- 8224839 TI - Perinatal lethality and defects in hindbrain development in mice homozygous for a targeted mutation of the zinc finger gene Krox20. AB - Krox20 is a zinc finger gene expressed in rhombomeres 3 and 5 during hindbrain development in vertebrates. Mice homozygous for a targeted mutation that deletes the majority of the Krox20 genes, including the zinc finger DNA-binding domain, died shortly after birth. The primary phenotype of the homozygous mutant animals was the loss of rhombomeres 3 and 5. This resulted in fusions of the trigeminal ganglion with the facial and vestibular ganglia, and of the superior ganglia of the glossopharyngeal and vagus nerves. These fusions resulted in a disorganization of the nerve roots of these ganglia as they entered the brain stem. These data demonstrate that Krox20 plays an essential role during development of the hindbrain and associated cranial sensory ganglia in mice. PMID- 8224840 TI - Migrations of the Caenorhabditis elegans HSNs are regulated by egl-43, a gene encoding two zinc finger proteins. AB - During embryonic development, the two Caenorhabditis elegans HSN motor neurons migrate from their birthplace in the tail to positions near the middle of the embryo. Here, we demonstrate that of all cells that undergo long-range migrations, only the HSNs are affected in animals that lack function of the egl 43 gene. We also show that egl-43 function is required for normal development of phasmid neurons, which are sensory neurons located in the tail. The egl-43 gene encodes two proteins containing zinc finger motifs that are similar to the zinc fingers of the murine Evi-1 proto-oncoprotein. Our genetic and molecular results suggest that egl-43 encodes two transcription factors and acts to control HSN migration and phasmid neuron development, presumably by regulating other genes that function directly in these processes. PMID- 8224841 TI - A switch from Myc:Max to Mad:Max heterocomplexes accompanies monocyte/macrophage differentiation. AB - Mad is a basic-helix-loop-helix-zipper protein that heterodimerizes with Max in vitro. Mad:Max heterodimers recognize the same E-box-related DNA-binding sites as Myc:Max heterodimers. However, in transient transfection assays Myc and Mad influence transcription in opposite ways through interaction with Max; Myc activates while Mad represses transcription. Here, we demonstrate that Mad protein is induced rapidly upon differentiation of cells of the myeloid lineage. The Mad protein is synthesized in human cells as a 35-kD nuclear phosphoprotein with an extremely short half-life (t1/2 = 15-30 min) and can be detected in vivo in a complex with Max. In the undifferentiated U937 monocyte cell line Max was found complexed with Myc but not Mad. However, Mad:Max complexes began to accumulate as early as 2 hr after induction of macrophage differentiation with TPA. By 48 hr following TPA treatment only Mad:Max complexes were detectable. These data show that differentiation is accompanied by a change in the composition of Max heterocomplexes. We speculate that this switch in heterocomplexes results in a change in the transcriptional regulation of Myc:Max target genes required for cell proliferation. PMID- 8224842 TI - Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. AB - The activity of c-Jun is regulated by phosphorylation. Various stimuli including transforming oncogenes and UV light, induce phosphorylation of serines 63 and 73 in the amino-terminal activation domain of c-Jun and thereby potentiate its trans activation function. We identified a serine/threonine kinase whose activity is stimulated by the same signals that stimulate the amino-terminal phosphorylation of c-Jun. This novel c-Jun amino-terminal kinase (JNK), whose major form is 46 kD, binds to a specific region within the c-Jun trans-activation domain and phosphorylates serines 63 and 73. Phosphorylation results in dissociation of the c-Jun-JNK complex. Mutations that disrupt the kinase-binding site attenuate the response of c-Jun to Ha-Ras and UV. Therefore the binding of JNK to c-Jun is of regulatory importance and suggests a mechanism through which protein kinase cascades can specifically modulate the activity of distinct nuclear targets. PMID- 8224843 TI - Cell cycle-regulated nuclear localization of MCM2 and MCM3, which are required for the initiation of DNA synthesis at chromosomal replication origins in yeast. AB - MCM2 and MCM3 are two genetically interacting and structurally related proteins essential for growth in Saccharomyces cerevisiae. Mutants defective in these proteins affect the stability of minichromosomes in general, but the severity of the defect is dependent on the autonomously replicating sequence (ARS) that drives the replication of that plasmid. In this paper we show by two-dimensional gel electrophoresis that the initiation of DNA synthesis at chromosomal replication origins is also reduced in frequency in these mutants. We show further that the nuclear and subnuclear localizations of the MCM2 and MCM3 proteins are temporally regulated with respect to the cell cycle. These proteins enter the nucleus at the end of mitosis, persist there throughout G1 phase, and disappear from it at the beginning of S phase. Once inside the nucleus, a fraction of the MCM2 and MCM3 proteins becomes tightly associated with DNA. The association of MCM2 and MCM3 with chromatin presumably leads to the initiation of DNA synthesis, and their subsequent disappearance from the nucleus presumably prevents reinitiation of DNA synthesis at replication origins. This temporally and spatially restricted localization of MCM2 and MCM3 in the nucleus may serve to ensure that DNA replication occurs once and only once per cell cycle. PMID- 8224844 TI - Extracellular cAMP is sufficient to restore developmental gene expression and morphogenesis in Dictyostelium cells lacking the aggregation adenylyl cyclase (ACA). AB - Cell movement and cell-type-specific gene expression during Dictyostelium development are regulated by cAMP, which functions both as an extracellular hormone-like signal and an intracellular second messenger. Previous data indicated that aca- mutants, which lack adenylyl cyclase activity, fail to aggregate and do not express cell-type-specific genes. We show here that overexpression of ACG, a constitutively active adenylyl cyclase, which in wild type cells is only expressed during spore germination, partially restores the coordination of cell movement and completely restores developmental gene expression. The aca- cells can also be induced to develop into viable spores by synergy with wild-type cells and, furthermore, form small but normal fruiting bodies, after a developmentally relevant regimen of stimulation with nanomolar cAMP pulses followed by micromolar cAMP concentrations. 2'-Deoxy cAMP, a cAMP analog that activates the cell-surface cAMP receptors but not cAMP-dependent protein kinase (PKA), also induces fruiting body formation as well as expression of prespore-specific and prestalk-enriched genes in aca- cells. Intracellular cAMP levels were not altered in aca- cells after stimulation with 2'-deoxy cAMP. Our data indicate that ACA is not required to provide intracellular cAMP for PKA activation but is essential to produce extracellular cAMP for coordination of cell movement during all stages of development and for induction of developmental gene expression. PMID- 8224845 TI - Activity of Wnt-1 as a transmembrane protein. AB - The product of the Wnt-1 proto-oncogene is a cysteine-rich glycoprotein that plays a crucial role in the development of the vertebrate central nervous system. Wnt-1 protein is secreted but remains associated with the cell surface and extracellular matrix. The function of Wnt-1 in several different biological settings can be carried out by cells that receive the Wnt signal from adjacent cells. Ectopic expression of Wnt-1 in certain mammary gland cell lines, such as C57MG, causes morphological transformation; C57MG cells can also be transformed by a paracrine mechanism when mixed with other cell types secreting Wnt-1 protein. To ask whether Wnt-1 protein can function while bound to the cell of origin, a variety of cell types were programmed to produce chimeric proteins containing the complete sequence of mature Wnt-1 protein fused to part or all of the transmembrane protein CD4 or CD8. The chimeras were found at the cell surface of transfected cells and did not appear to be proteolytically processed. In autocrine and paracrine transformation assays with C57MG cells and in an axis induction assay in Xenopus laevis embryos, the Wnt-1/CD4 or CD8 fusions retained significant activity, as did a secreted chimera containing the CD8 extracellular domain but lacking the transmembrane domain. However, a chimera lacking a spacer between the Wnt-1 and the transmembrane domains was weakly active and only in autocrine transformation. These results show that tethering Wnt-1 to the cell surface still allows Wnt-1-mediated cell-to-cell signaling. PMID- 8224846 TI - In vivo cooperation between introns during pre-mRNA processing. AB - In higher eukaryotes the large number of introns present in most genes implies that the pre-mRNA processing machinery should be efficient and accurate. Although this could be achieved at the level of each intron, an attractive alternative would be that interactions between introns improve the performance of this machinery. In this study we tested this hypothesis by comparing the processing of transcripts of the tumor necrosis factor beta gene, which differ only by their number of introns. We took advantage of the ordered splicing of the three introns present in this gene to design constructs that should generate, as primary transcripts, molecules that are normally produced by splicing. We established that the apparent splicing rate of intron 3 is increased 2.5- and 3.5-fold by the presence of one or two other introns on the primary transcript, respectively. Similarly, the apparent splicing rate of intron 2 is increased by the presence of intron 1. As these effects involve the splice sites of the upstream intron, these observations support the existence of cooperative interactions between introns during pre-mRNA processing. PMID- 8224847 TI - SV40 early-to-late switch involves titration of cellular transcriptional repressors. AB - We have purified factors from HeLa cell nuclear extracts that bind to the transcriptional initiation site of the SV40 major late promoter (SV40-MLP). The resulting fraction consists predominantly of three proteins, collectively called initiator-binding protein of SV40 (IBP-s) with electrophoretic mobilities of approximately 45-55 kD. Gel mobility-shift and DNase I-protection analyses indicate that each of these three proteins associates with high affinity to sequences located at the initiation site and 55 bp downstream of it. IBP-s binding sites with lower affinities are located at +5 and +30. Addition of purified IBP-s to a cell-free transcription system represses transcription from the SV40-MLP, but not the SV40 early promoter. SV40 mutants lacking the two strongest IBP-s-binding sites (1) are not repressed by the addition of IBP-s in vitro, (2) overproduce late RNA (relative to wild-type SV40) at low, but not high, template copy number in vitro, and (3) exhibit increased levels of late RNA at early, but not late, times after transfection into CV-1 cells. Therefore, IBP s is a cellular repressor of transcription of the SV40-MLP that may, in large part, be responsible for the replication-dependent component of the early-to-late shift in SV40 gene expression. Partial amino acid sequence data obtained from the approximately 55-kD component of IBP-s indicate that it is hERR1, an orphan member of the steroid-thyroid hormone receptor superfamily. These findings suggest simple molecular mechanisms by which hormones may modulate expression of viral late genes. We speculate that activation of expression of the late genes of other viruses may occur by similar mechanisms. PMID- 8224848 TI - A single cDNA, hTFIIA/alpha, encodes both the p35 and p19 subunits of human TFIIA. AB - TFIIA is a transcription factor that, by interacting with the TATA-binding subunit (TBP) of TFIID, modulates transcription initiation by RNA polymerase II in vitro. By use of a mobility shift assay, TFIIA was purified from HeLa cells as a complex of 35-, 19-, and 12-kD subunits. Oligonucleotides were used to isolate a human cDNA clone, hTFIIA/alpha, which encodes a 55-kD protein with homology to the product of the yeast gene TOA1. The open reading frame of hTFIIA/alpha contains peptide sequences obtained from both the p35 and p19 subunits of natural human TFIIA, and thus encodes these two subunits. Consistent with this, antiserum raised against the 55-kD hTFIIA/alpha-encoded protein reacted with both the p35 and p19 subunits of natural TFIIA, and the recombinant protein could functionally replace those subunits in a mobility shift assay with renatured p12. An efficient affinity purification for natural human TFIIA was suggested by the sequence of the hTFIIA/alpha protein and demonstrated biochemically. Finally, transcription from the adenovirus major late promoter was greatly reduced in nuclear extracts depleted with anti-TFIIA/alpha serum and was restored to original levels by the readdition of purified human TFIIA. PMID- 8224849 TI - Drosophila TFIIA-L is processed into two subunits that are associated with the TBP/TAF complex. AB - The basal factor TFIIA has been shown to act early during initiation in both the mammalian and yeast transcription systems, but a TFIIA-like activity has not been identified in Drosophila. While characterizing the Drosophila TFIID complex, we discovered that a 30-kD protein that cofractionated with dTFIID was homologous to the previously identified, large subunit of yeast TFIIA. Here, we report the cloning and biochemical characterization of Drosophila TFIIA-L. Coimmunoprecipitation studies with anti-dTBP, anti-dTFIIA-L, and anti-TAF antibodies indicated a tight association of the endogenous dTFIIA and dTFIID. However, dTFIIA could be dissociated from dTFIID under conditions that did not elute the TAFs, and the eluted material had mobility shift and transcriptional activities associated with TFIIA. Peptide sequence and Western analysis with antibodies raised against the amino- and carboxy-terminal portions of recombinant dTFIIA-L revealed that a precursor 48-kD species was cleaved in vivo, giving rise to the 30- and 20-kD subunits of dTFIIA that remain associated with each other and with dTFIID. Protein-protein interaction assays identified dTBP and dTAFII110 as targets for binding TFIIA in the TFIID complex. These results suggest that TFIIA may form a specific complex with both TAFs and other components of the transcriptional machinery during formation of the initiation complex. PMID- 8224850 TI - Isolation of a cDNA encoding the largest subunit of TFIIA reveals functions important for activated transcription. AB - Transcription factor IIA has been shown to interact with the TATA-binding protein and to act early during preinitiation complex formation. The human factor is composed of three subunits (alpha, beta, gamma). A human cDNA clone encoding the largest subunit of TFIIA (alpha) was isolated. The recombinant alpha polypeptide, together with the beta and gamma subunits, was capable of reconstituting TFIIA activity. Studies using antibodies raised against recombinant alpha polypeptide demonstrate that TFIIA can be an integral component of the preinitiation complex. We demonstrate that TFIIA not only interacts with TBP but also can associate with the TFIID complex. Functional assays establish that TFIIA has no apparent role in basal transcription but plays an important role in activation of transcription. Interestingly, amino acid sequence analyses of the beta-subunit demonstrate these residues to be entirely contained within the carboxyl terminus of the cDNA clone encoding the alpha-subunit. PMID- 8224851 TI - Variable X chromosome inactivation patterns in near-tetraploid murine EC x somatic cell hybrid cells differentiated in vitro. AB - For the cytogenetic study of X chromosome inactivation as an X chromosome dosage compensation mechanism, we isolated a number of XXXX, XXX, and XXY near tetraploid mouse hybrid cell clones by fusing XX or XO embryonal carcinoma cells with lymphocytes carrying a structurally altered X chromosome(s). The inactive X chromosome from the female lymphocyte was reactivated in these hybrid clones which retained embryonal carcinoma morphology so far as they were cultured on the collagen-coated plastic surface in the medium supplemented with leukemia inhibitory factor (LIF) and betamercaptoethanol (BME). Some of these clones developed balloon-like cystic embryoid bodies when they were allowed to form cell aggregates in medium without LIF and BME in bacteriological petri dishes to which they do not adhere. X chromosome inactivation occurring during this process detected by the incorporation of 5-bromodeoxyuridine did not conform to the expected pattern leaving two X chromosomes active in every tetraploid cells. This may suggest either that the X-inactivation mechanism evolved primarily, for the diploid cell is unable to deal with tetraploid conditions efficiently, or that the present system of in vitro differentiation represents an anomalous situation never encountered in vivo. PMID- 8224852 TI - Genetics and biology of the sawfly, Athalia rosae (Hymenoptera). Review. AB - Hymenopteran insects are a unique group of animals in which arrhenotokous reproduction (haploid males develop from unfertilized eggs) is a rule. Males produce sperm through a non-reductional maturation division. A sawfly species, Athalia rosae ruficornis Jakovlev Tenthredinidae, Symphyta, Hymenoptera) has introduced as a new experimental material for studies on genetics and developmental biology. Basic features relating to the potential usefulness of the species in elucidating some to the important genetic and developmental biological problems are described. PMID- 8224853 TI - Genetics of food preference in Drosophila sechellia. I. Responses to food attractants. AB - To reveal the genetic mechanism of host selection in a monophagous fruit fly Drosophila sechellia, olfactory responses and oviposition preferences of this species were compared with those of closely related polyphagous species, D. simulans and D. melanogaster. Adult flies of D. sechellia were strongly attracted to the ripe fruit of Morinda citrifolia which is known to be the sole breeding site of this species. They were also attracted to the odor of n-caproic acid which is contained in the ripe fruit of M. citrifolia and is presumably responsible for the characteristic odor of the fruit. In contrast, D. simulans and D. melanogaster showed a strong repulsion to n-caproic acid. In parallel with the olfactory responses, D. sechellia females laid eggs preferentially on a medium containing n-caproic acid, to which the other two species showed an aversion. Genetic analyses using the hybrid progeny between D. sechellia and D. simulans suggested that the species differences in these behaviors are controlled by gene(s) located on the second chromosome. PMID- 8224854 TI - In search of new mutants in cell-signaling systems of the nematode Caenorhabditis elegans. Review. AB - Development of multicellular organisms is controlled mainly by cell-signaling systems. In this review I first discuss methods of genetic analysis and properties of mutants of cell-signaling systems in general and in the nematode C. elegans. Then, I describe two of our approaches to isolating new mutants in cell signaling of C. elegans. The first approach is to select for mutants that have the same visible phenotype as those in known cell-signaling genes. In a survey of larval lethal mutations we found that there are quite a few mutants in which the inner surface of the body wall is detached from the outer surface of the intestine. Some of them map in genes that are known to act in cell-signaling systems in vulval induction or sex myoblast migration, which are not essential to the growth and survival of C. elegans. Therefore, we think many of the mutations of the above phenotype disrupt cell-signaling in an unidentified essential function, and also cell-signaling in the non-essential functions. The second approach is to isolate mutants resistant to a drug expected to disturb cell signaling. As the drug we have chosen sodium fluoride, which depletes calcium ion, activates G-proteins and inactivates some phosphatases. The mutants are grouped into two classes (three and two genes, respectively) according to degree of fluoride-resistance and growth rate of larvae. Although there is so far no direct evidence that these mutants are related to cell-signaling, they show complex epistasis that can be explained by a model consisting of a cell-signaling pathway. PMID- 8224856 TI - Meiotic mutations from natural populations of Drosophila melanogaster. AB - Two meiotic genes from natural populations are described. A female meiotic mutation, mei(1)g13, mapped to 17.4 on the X chromosome, causes nondisjunction of all homologs except for the fourth chromosomes. In addition, it reduces recombination by 10% in the homozygotes and causes 18% increased recombination in the heterozygotes. A male meiotic mutation, mei-1223m144, is located on the third chromosome. Although this mutation causes nondisjunction of all chromosomes, each chromosome pair exhibits a different nondisjunction frequency. Large variations in the sizes of the premature sperm heads observed in the homozygotes may reflect irregular meiotic pairing and the subsequent abnormal segregation, resulting in aneuploid chromosome complements. PMID- 8224855 TI - Positive and negative signaling mechanisms in the regulation of photoreceptor induction in the developing Drosophila retina. Review. AB - An ommatidium of a Drosophila compound eye contains eight photoreceptor cells, R1 R8. The fates of the photoreceptors are determined exclusively by inductive interactions between neuronal precursors in the cell cluster from which the ommatidium is formed. R7 induction has been extensively analysed at the molecular level. Activation of a membrane receptor tyrosine kinase (Sevenless) in the R7 precursor by a ligand (Bride of sevenless) present on the surface of R8 triggers a transduction cascade mediated by Ras, establishing the R7 fate of this cell. Other Sev-expressing cells are prevented from taking on the R7 fate by several different mechanisms. Pokkuri-mediated repression represents one such regulatory mechanism. The positive and negative signaling pathways operating in the fate determination of other photoreceptor cells are also discussed. PMID- 8224857 TI - Hybrid lethal systems in the Drosophila melanogaster species complex. AB - Lethal phases of the hybrids between Drosophila melanogaster and its sibling species, D. simulans are classified into three types: (1) embryonic lethality in hybrids carrying D. simulans cytoplasm and D. melanogaster X chromosome, (2) larval lethality in hybrids not carrying D. simulans X, and (3) temperature sensitive pupal lethality in hybrids carrying D. simulans X. The same lethal phases are also observed when either of the two other sibling species, D. mauritiana or D. sechellia, is employed for hybridization with D. melanogaster. Here, we describe genetic analyses of each hybrid lethality, and demonstrate that these three types of lethality are independent phenomena. We then propose two models to interpret the mechanisms of each hybrid lethality. The first model is a modification of the conventional X/autosome imbalance hypothesis assuming a lethal gene and a suppressor gene are involved in the larval lethality, while the second model is for embryonic lethality assuming an interaction between a maternal-effect lethal gene and a suppressor gene. PMID- 8224858 TI - Hotspots of meiotic recombination in the mouse major histocompatibility complex. AB - Meiotic recombination is not random in the proximal region of the mouse major histocompatibility complex (MHC). It is clustered at four restricted positions, so-called hotspots. Some of the MHC haplotypes derived from Asian wild mice enhance recombination at the hotspots in genetic crosses with standard MHC haplotypes of laboratory mouse strains. In particular, the wm7 haplotype derived from Japanese wild mouse indicated an approximately 2% recombination frequency within a 1.2 kb fragment of DNA in the interval between the Pb and Ob genes. Interestingly, this enhancement of recombination was observed only in female meiosis but not in male meiosis. Mating experiments demonstrated that the wm7 haplotype carries a genetic factor in the region proximal to the hotspot, which instigates recombination. In addition, the wm7 haplotype has a genetic factor located in the region distal to the hotspot, which suppresses recombination. From the molecular characterization of the two hotspots located in the Eb gene and the Pb-Ob interval, it appeared that there are several common molecular elements, the consensus of the middle repetitive MT-family, TCTG or CCTG tetramer repeats, and the solitary long terminal repeat (LTR) of mouse retrovirus. PMID- 8224860 TI - Lack of site-specific recombination between mitochondrial genomes of petite mutants of yeast. AB - Previous work from our laboratory showed that mitochondrial (mt) genomes, with tandem repeat units, from spontaneous, cytoplasmic petite mutants of Saccharomyces cerevisiae do not exhibit site-specific recombination in petite x petite crosses [Rayko et al., Gene 63 (1988) 213-226]. Here, we have extended and confirmed these observations by studying other crosses of petites with tandem repeat units, as well as crosses in which one of the parents was, instead, an unstable petite, a-15/4/1, having a palindromic mt genome. In no case was site specific recombination of the parental mt genomes observed. Progeny cells harbored mt genomes derived from either one or both of the two parents, as shown by analysis of restriction fragments. In the case of biparental inheritance, extensive subcloning of the diploids showed that this was due to a persistent heteroplasmic state and not to intermolecular recombination. The 'new' restriction fragments present in the mt DNA from some diploids were shown to be derived from the unstable parental genome, a-15/4/1, by a secondary excision process. Lack of site-specific recombination is, therefore, not only a feature of crosses involving petite genomes made up of tandem repeat units, but also of crosses in which one parental genome consists of inverted repeats and frequently originates secondary petite genomes formed by tandem repeats. Previous reports of mt recombination in petite mutants are discussed in light of these results. PMID- 8224859 TI - W-heterochromatin of chicken; its unusual DNA components, late replication, and chromatin structure. AB - About 65% of DNA in the chicken W chromosome has been shown to consist of XhoI and EcoRI family repetitive sequences. These sequences showed remarkable delay in the electrophoretic mobility at low temperature on a polyacrylamide gel. Three dimensional structures of the 0.7-kb XhoI and the 1.2-kb EcoRI family repeating units were estimated to be irregular solenoids using a computer program based on wedge angles of all the 16 dinucleotide steps. Fluorescence in situ hybridization demonstrated that these two family sequences were localized in a major heterochromatic body in an interphase nucleus. Incorporation of bromodeoxyuridine into the W chromosome in the synchronous culture of MSB-1 cells occurred about 1 h later than the peak of S phase. The chromatin structure formed along XhoI and EcoRI family sequences was suggested to be different from the total chromatin or chromatin containing the beta-actin gene sequence in that the linker DNA lengths of the former were significantly longer. Fractionation of the HaeIII-digested MSB 1 nuclei yielded a chromatin fraction in which XhoI family sequences were partially enriched. Several DNA-binding proteins showing higher affinity for the XhoI family sequence were present in this fraction. PMID- 8224861 TI - The Saccharomyces cerevisiae MTS1 gene encodes a putative RNA-binding protein involved in mitochondrial protein targeting. AB - Most proteins present in the mitochondrion are nuclear encoded, and are directed to the organelle by virtue of a targeting sequence at the N terminus of the precursor protein. Mitochondrial (mt) protein targeting appears to require several accessory proteins that recognise mt precursors both in the cytoplasm and at the mt surface. We describe here the use of yeast genetics to identify a protein that is required for mt protein targeting. Two yeast mutants (mts1 and mts2) were isolated as extragenic suppressors of a known targeting defect in the presequence of the beta-subunit of ATP synthase. We have cloned and sequenced the wild-type allele of one of these genes (MTS1) and shown that it encodes a member of a family of RNA-binding proteins that is essential for growth. PMID- 8224862 TI - Characterisation, cloning and integrative properties of the gene encoding urate oxidase in Aspergillus nidulans. AB - A number of mutations have been obtained which define the structural gene (uaZ) coding for urate oxidase in linkage group I of Aspergillus nidulans. This gene has been cloned by transformation of a uaZ- null mutant. A chromosome I/VIII translocation which splits the gene has been defined both genetically and physically. All known mutations are contained in a 1-kb fragment, itself contained in the probe which recognizes a 1.2-kb inducible message. Plasmids carrying uaZ show a strict bias towards homologous recombination in transformation experiments. PMID- 8224863 TI - Heterologous protein secretion directed by a repressible acid phosphatase system of Aspergillus niger. AB - A new expression-secretion system of Aspergillus niger which directs the secretion of heterologous proteins is described. The promoter and signal peptide encoding region of the phosphate-repressible aphA gene of A. niger, when fused to the coding region of the human interferon alpha 2 (hIFN alpha 2)-encoding gene (hIFN alpha 2), drives the expression of this gene and the secretion of the hIFN alpha 2 protein. Synthesis of hIFN alpha 2 in either A. niger or A. nidulans transformants carrying these constructs was regulated by inorganic phosphate (Pi) present in the medium, so that derepression of heterologous protein expression can be attained by lowering Pi concentration. PMID- 8224864 TI - The requirement for subunit interaction in the production of Penicillium chrysogenum acyl-coenzyme A:isopenicillin N acyltransferase in Escherichia coli. AB - Subunit interaction in the formation of active acyl-coenzyme A:isopenicillin N acyltransferase (AT) has been investigated. Various AT derivatives were produced from altered Penicillium chrysogenum penDE genes placed in Escherichia coli expression systems. The regions of penDE encoding the alpha (11 kDa) and beta (29 kDa) AT subunits were separated at the DNA level by linker insertion at the region encoding Gly102/Cys103. Synthesis of AT from the resulting two-cistron mRNA resulted in active alpha,beta-heterodimeric recombinant AT (reAT), containing subunits of 11 and 29 kDa (similar to wild-type AT). Complete separation of the alpha and beta subunits was performed by placing the region of penDE encoding each subunit on different plasmids. Production of either subunit in the absence of the other did not form active reAT. However, cotransformation of E. coli with two plasmids, each encoding a different AT subunit, produced reAT having acyl-coenzyme A:6-aminopenicillanic acid (acyl-CoA:6-APA) AT activity. Mutation of penDE replacing Thr105 with Asn resulted in inactive and uncleaved reAT. Coexpression of this mutant penDE with a penDE derivative encoding the beta subunit in E. coli produced acyl-CoA:6-APA AT activity. These results suggest that the formation of reAT involves cooperative folding events between the subunits. In vitro transcription/translation was used to determine the origin of the AT hydrolase activity that cleaves the 40-kDa precursor polypeptide. The appearance of a 29-kDa protein (and presumably the corresponding 11-kDa protein, although not observable) from the 40-kDa in vitro translated protein provides further evidence that AT hydrolysis is an autocatalytic event. PMID- 8224865 TI - Stable, selectable, integrative DNA transformation in Physarum. AB - The Physarum polycephalum actin promoter, PardC, can drive transient expression of heterologous genes in Physarum amoebae. The hph gene, encoding hygromycin (Hy) phosphotransferase, can confer resistance to Hy on a broad spectrum of organisms. When PardC is translationally fused to hph and transformed into yeasts on high copy-number vectors, the yeasts become Hy resistant (HyR), showing that PardC-hph is a functional, selectable genetic element. To establish a stable transformation system for Physarum, we electroporated plasmids bearing PardC-hph into Physarum amoebae and then selected for HyR transformants. We show that HyR amoebae arise upon the stable integration of PardC-hph into the nuclear genome in single copy. These results establish a transformation system that can be used to add plasmid borne genetic information to Physarum. PMID- 8224866 TI - Silicon-responsive cDNA clones isolated from the marine diatom Cylindrotheca fusiformis. AB - In organisms ranging from single-celled algae to mammals, including humans, silicon is essential for, and actively participates in, a variety of life processes. It has become clear that silicon (i) acts as a metabolite affecting a variety of cellular processes, and (ii) regulates gene expression. However, the mechanisms by which silicon (i.e., Na2SiO3.9H2O, in the present study) acts are not clear, due to inherent methodological difficulties. As part of our program to understand how silicon acts in biological systems, we present the first isolation of cDNA clones derived from silicon-responsive mRNAs, from the marine diatom Cylindrotheca fusiformis. We distinguish between clones responding only to silicon starvation and replenishment, and those also responding to other cellular conditions. Some of the clones can be identified by similarity to other genes, and should be useful as probes to isolate genes from other organisms. Isolation of these clones provides the means to (i) identify metabolic pathways affected by silicon, and (ii) investigate the mechanism(s) of silicon-regulated gene expression. PMID- 8224867 TI - Abundant mRNAs in the squid light organ encode proteins with a high similarity to mammalian peroxidases. AB - A library derived from mRNA in the bacterial light organ of the squid, Euprymna scolopes, contained an unexpectedly high proportion of cDNAs that encode proteins with approximately 30% similarity to a family of mammalian peroxidases (PO) including myelo-PO, eosinophil PO, and thyroid PO (donor:hydrogen-peroxide oxidoreductase; EC 1.11.1.7). Two nearly full-length cDNAs were determined to encode putative PO of nearly 93 kDa each that are 97% identical in amino acid sequence to each other. Each contains four potential glycosylation sites, and His416, believed to be within the active site of the human PO, is conserved in the putative PO from the squid light organ. The mRNAs for the putative squid PO were approximately 250 times more abundant in the tissue housing the bacterial symbiont than in the ocular lens or mantle and were undetectable in the light organ lens. By analogy with the bacteriocidal function of PO in mammalian neutrophils, the putative squid PO may be important for modulating or limiting the population of bacteria within the light organ. The possibility that the squid light organ contains a high concentration of PO raises the possibility that the light organ lens is under oxidative stress, providing a possible rationale for the recruitment of its aldehyde dehydrogenase-like crystallin. PMID- 8224868 TI - Cloning and sequence analysis of the rat tumor necrosis factor-encoding genes. AB - We have isolated an approximately 22-kb TNF locus (encoding tumor necrosis factor) from a rat genomic library and sequenced the 7105-bp fragment that comprises the TNF-alpha and TNF-beta genes, including their flanking sequences. The two genes are tandemly arranged with TNF-beta 5' to TNF-alpha and separated by approximately 1.1 kb of intergenic space, and each gene consists of four exons and three introns, similar to those of the other species examined thus far. Comparison analysis showed that the rat TNF have high sequence homology with the mouse TNF (TNF-alpha, 86.5%; TNF-beta, 89.3%) and relatively low homology with the human, rabbit, and porcine TNF. The upstream sequence of rat TNF-alpha contains a number of sequence motifs implicated in the expression and regulation of eukaryotic genes, including binding sites for the transcription factors Sp-1, Ap-2, IFN.1 and NF-kappa B. The possible significance of potential regulatory sequence elements found in the rat TNF-alpha in the context of transcriptional regulatory mechanisms is discussed. PMID- 8224869 TI - Cloning and analysis of a cDNA encoding mammalian arginyl-tRNA synthetase, a component of the multisynthetase complex with a hydrophobic N-terminal extension. AB - In mammalian cells, the nine aminoacyl-tRNA synthetases (aaRS) specific for the amino acids (aa) Glu, Pro, Ile, Leu, Met, Gln, Lys, Arg and Asp are associated within a multienzyme complex. Arginyl-tRNA synthetase (ArgRS) is characterized by the occurrence of two structurally distinct forms of that enzyme: a complexed (approximately 74 kDa) and a free (approximately 60 kDa) form. The cDNA encoding the 74-kDa species of ArgRS from Chinese hamster ovary cells has been isolated and sequenced. The deduced aa sequence shows 38% identity to the homologous bacterial enzyme but displays an N-terminal polypeptide extension composed of 73 aa, which is absent in the free form of mammalian ArgRS. Two regions of this extension are predicted to be alpha-helical, leading to the clustering of Leu and Ile residues on one side of the helices. This suggests that the N-terminal domain is involved in the assembly of the 74-kDa species of ArgRS within the multisynthetase complex through hydrophobic interactions. By using the isolated cDNA, a Northern blot analysis showed a single mRNA species. Thus, there is a possibility that the free and complexed forms of ArgRS are encoded by the same gene. PMID- 8224870 TI - Distribution of bent DNA structures in the fission yeast centromere. AB - To gain a clue as to the functional significance of DNA curvature, we experimentally characterized the distribution of bent DNA structures throughout the 35-kb cen1 sequence, one of the isolated functional centromeric DNA of the fission yeast, Schizosaccharomyces pombe. It was revealed that a relatively large central portion of cen1, covering a 2.2-kb DNA sequence, displays a remarkable DNA curvature. PMID- 8224871 TI - Sequence and expression of a gene encoding a ribosomal protein S4 homolog from Drosophila melanogaster. AB - We describe a Drosophila melanogaster cDNA clone encoding an amino acid (aa) sequence 75% identical to human ribosomal protein S4 (RPS4). The D. melanogaster RPS4 has 260 aa, if the NH2-terminal methionine is removed after translation of the mRNA. The mRNA for the protein is about 1 kb in length and is detected throughout the developmental stages tested (i.e., embryo, larva, pupa and adult). The cDNA clone hybridizes to two sites on the X chromosome, 1B1-2 and 3A3-6. PMID- 8224872 TI - Sequence of the gene coding for ribosomal protein S8 of Xenopus laevis. AB - We present here the cloning and the entire sequence of one of the two gene copies coding for ribosomal protein (r-protein) S8 in Xenopus laevis (corresponding to r protein S7 in rat) and its flanking regions. The S8a gene contains seven exons and six introns for a total length of about 12,700 bp coding for a mRNA of 663 nucleotides (nt) plus a poly(A) tail. Mapping of the 5' end of the gene has shown that the transcription start point is located in a pyrimidine-rich tract, as has been observed for all r-protein-encoding genes of X. laevis and other vertebrates so far characterized. A computer analysis of the S8a sequence has revealed the presence of a 220-nt sequence repeated, with some variations, once in each of the six introns. RNA analysis by hybridization with oligo probes specific for the two gene copies coding for r-protein S8 has demonstrated that the two of them are expressed at similar levels both in oocytes and in embryos. PMID- 8224873 TI - Characterization of the 5' end of the mouse Ent gene encoding the basement membrane protein, entactin. AB - Entactin (Ent) is a 150-kDa basement membrane glycoprotein that forms a complex with laminin and also binds to type-IV collagen. For the initial characterization of the 5' end of the mouse Ent gene, we isolated genomic clones that encompass the first three exons of the gene. A 3.5-kb EcoRI fragment at the 5' end of the gene was sequenced and shown to contain 1226 bp of the 5'-flanking DNA, the 260 bp first exon, and 2060 bp of the first intron. The second exon is separated from the first exon by an intron that spans greater than 18 kb. The major transcription start point (tsp) was mapped to 35 nucleotides upstream from the translation start codon. The 5'-flanking DNA contains a putative TATA box, two CAAT boxes and two GC boxes. Nine potential AP-2- and two potential AP-1-binding sites were found in the upstream region and the first intron. The first 248 bp of the promoter region are G+C rich, and they are 65% identical to the promoter of human Ent. Fragments of the 5'-flanking DNA linked to a reporter gene were shown to have promoter activity in transient transfection assays. PMID- 8224874 TI - Transcription factor RFX1 helps control the promoter of the mouse ribosomal protein-encoding gene rpL30 by binding to its alpha element. AB - The factor that binds to the most upstream element (alpha) of the mouse rpL30 promoter was identified as RFX1, a novel 105-kDa protein that recognizes an important element of MHC class-II promoters. Identification was based on competition between rpL30 alpha and an RFX1-binding site for nuclear protein complex formation and on the ability of RFX1 antibody to supershift the electrophoretic mobility of the DNA-protein complexes. A mutation in the alpha element that abolished its interaction with RFX1 reduced rpL30 promoter activity to about 43% of the wild-type level, indicating that RFX1 plays an important role in determining the strength of the rpL30 promoter. A search of a eukaryotic promoter database revealed candidate RFX1-binding sites in a variety of other promoters, suggesting that this protein may be implicated in the transcriptional regulation of a wide variety of genes. PMID- 8224875 TI - Human ribosomal protein L37a: cloning of the cDNA and analysis of differential gene expression in tissues and cell lines. AB - A cDNA corresponding to human ribosomal protein L37a (hL37a) was obtained by screening a SHSY5Y neuroblastoma library. The full-length cDNA contained 366 nucleotides (nt) in addition to a poly(A) tail. The pyrimidine-rich sequence, CTTTCT, that is common to many ribosomal protein-encoding cDNAs was present at the 5' terminus. The nt sequence displayed 85% identity with rat L37a (rL37a) cDNA. The predicted protein contains 92 amino acids with a M(r) of 10,277, is highly basic, and has 100% sequence identity with rL37a. A putative zinc-finger domain is present in the central region of the protein. Human lymphocytes and several human cell lines express hL37a mRNA at significantly higher levels than the rat cell lines and rat tissues tested. The hL37a gene does not contain introns. PMID- 8224876 TI - Human transforming growth factor alpha: sequence analysis of the 4.5-kb and 1.6 kb mRNA species. AB - Two transforming growth factor alpha (TGF alpha) mRNA species with the apparent sizes of 4.5 and 1.6 kb were identified in all human cell lines analysed. The cDNA corresponding to the 4.5-kb species was entirely sequenced, revealing the presence of a 3'-untranslated region (3'-UTR) of 3571 nucleotides, which contained several potential polyadenylation signals. Our results indicate that the 1.6-kb species is derived from the same precursor by alternative polyadenylation. In addition, we present evidence suggesting that TGF alpha specific mRNAs could be initiated from transcription start points (tsp) located upstream from the tsp previously identified by Jakobovitz et al. [Mol. Cell. Biol. 8 (1988) 5549-5554]. PMID- 8224877 TI - Polymorphism in the multi-phosphorylation domain of the human neurofilament heavy subunit-encoding gene. AB - The C-terminal region of the human neurofilament heavy subunit (NEFH) contains a unique functional domain consisting of 43 repeat motifs of the amino acids (aa) Lys-Ser-Pro (KSP) with either 3- or 5-aa spacers in between. Past studies have demonstrated that the serine in these KSP motifs can be phosphorylated, resulting in heavy phosphorylation of this domain. Recent studies provide strong evidence for a role of neurofilament phosphorylation in the establishment of neurofilament density and axonal caliber. Since it may be hypothesized that mutations in the phosphorylated region are a basis for neuropathological conditions, and since regions of the human genome containing repeat motifs have been demonstrated to be significantly polymorphic, we undertook to identify and characterize polymorphism in this region of the human NEFH gene. We were able to identify an allelic variant of a slightly larger molecular size, containing an additional KSP phosphorylation motif. The variant form of NEFH displays Mendelian inheritance and has a widespread population distribution. In addition, we also identified a point mutation in one individual which would result in a Pro-->Leu substitution in one of the repeat motifs. PMID- 8224878 TI - The cDNA sequence encoding human protein kinase C-zeta. AB - A 1779-bp complementary DNA (cDNA) that encodes protein kinase C-zeta (PKC-zeta) has been isolated from a human frontal cortex library using traditional plaque screening methods and PCR screening. The deduced 592-amino-acid sequence of the human PKC-zeta clone has a 95-96% identity to those deduced from the previously described rat and mouse PKC-zeta clones. PMID- 8224879 TI - Identification of a fowl adenovirus gene with sequence homology to the 100K gene of human adenovirus. AB - The nucleotide sequence of a region of the fowl adenovirus serotype 10 (FAV-10) genome, located between map units 57.3 and 62.6, has been determined. This sequence contains the entire coding region of the FAV equivalent to the 100K gene of human adenovirus (HAV). The FAV 100K polypeptide was found to consist of 798 amino acids with a deduced M(r) of 89,023. PMID- 8224880 TI - Mutations in the gene encoding the replication-initiation protein of plasmid RK2 produce elevated copy numbers of RK2 derivatives in Escherichia coli and distantly related bacteria. AB - Mini-replicons of the broad-host-range plasmid RK2 with increased copy number (cn) due to mutations in the gene encoding the essential replication initiation protein TrfA are described. The cn of these derivatives have been determined in Escherichia coli, Pseudomonas aeruginosa and Agrobacterium tumefaciens and were found to be elevated in all three bacterial hosts. One of the cn mutations was introduced into the intact 60-kb RK2 plasmid by homologous recombination in vivo, resulting in an approximately twofold cn increase. The expression of trfA from this mutant RK2 plasmid did not respond to the cn change as predicted by a simple transcription rate-limitation, replication control model. Implications for the model of RK2 replication control and the potential use of mutant RK2 mini replicons as high-copy broad-host-range gene cloning vectors are discussed. PMID- 8224881 TI - Gene sequence, overproduction, purification and determination of the wild-type level of the Escherichia coli flagellar switch protein FliG. AB - The flagellar motor switch in Escherichia coli and Salmonella typhimurium controls swimming behavior by regulating the direction of flagellar rotation. The switch is a complex apparatus composed of at least three proteins--FliG, FliM and FliN. During chemotactic behavior, the switch responds to signals transduced by the chemotaxis sensory signaling system. CheY, the chemotaxis response regulator, is thought to act directly on the switch to induce tumbles in the swimming pattern, but physical interaction of CheY and switch proteins has not been shown. We have undertaken this work to develop the molecular tools to investigate CheY binding to switch proteins, as well as to understand more about the structure and function of the switch. We present here the sequences of the fliG gene and its protein product, the engineering and amplification of fliG by the polymerase chain reaction (PCR) and its subcloning, and the overproduction, purification and determination of the wild-type (wt) level of the FliG protein. The sequence data revealed a 91.8% amino acid (aa) identity between E. coli and S. typhimurium FliG. Engineering and amplifying fliG by PCR allowed convenient cloning into an efficient expression vector. FliG was successfully overproduced and purified to > 98% purity. Polyclonal antibodies (Ab) were generated against purified FliG and used in quantitative Western blots to determine that the wt expression level of fliG results in about 3700 FliG copies per cell. Purified FliG and anti-FliG Ab will be useful for direct biochemical analyses of CheY-switch protein interaction. PMID- 8224882 TI - Cloning and sequencing of the hemE gene encoding uroporphyrinogen III decarboxylase (UPD) from Escherichia coli K-12. AB - Among the photoresistant revertants of the visA-deleted (hemH-deleted) strain of Escherichia coli K-12, three mutants defective in the hemE gene encoding uroporphyrinogen III decarboxylase (UPD) were identified. Using one of the mutants, we cloned and sequenced the hemE of E. coli. We found an open reading frame of 353 codons, which encoded a predicted amino acid (aa) sequence that exhibited a high degree of homology over its entire length to the aa sequence of UPD from humans and other organisms. This hemE was located at 90.3 min near the hupA gene on the linkage map of the E. coli chromosome. PMID- 8224883 TI - Characterization of the prtA and prtB genes of Erwinia chrysanthemi EC16. AB - Two tandem metalloprotease-encoding structural genes, prtA and prtB, were sequenced from Erwinia chrysanthemi EC16. These were highly homologous to previously reported genes from the same bacteria, as well as to three other metalloprotease-encoding genes from enteric bacteria. The three tandem prt structural genes from strain EC16 were closely linked to a cluster of genes previously found to be essential for extracellular secretion of the metalloproteases. PMID- 8224884 TI - Cloning and mapping of the Bacillus subtilis locus homologous to Escherichia coli ent genes. AB - Here, we report the cloning of a 3.5-kb HindIII fragment of chromosomal Bacillus subtilis DNA carrying at least two open reading frames exhibiting significant homology with entA and entE of Escherichia coli. This B. subtilis ent locus was mapped at about 41 degrees. Its inactivation did not cause any detectable phenotype. PMID- 8224885 TI - Sequence and characterization of an insertion sequence, IS711, from Brucella ovis. AB - The nucleotide (nt) sequence of a previously discovered insertion in Brucella ovis was determined and found to have the hallmarks of an insertion sequence (IS). The element, designated IS711, of 842 bp, is similar in G + C content to that of the Brucella genome and is bounded by 20-bp imperfect inverted repeats (IR). The element appears to duplicate the nt TA of a consensus target site, YTAR (R, purines; Y, pyrimidines). When the complete nt sequence of four elements and 300 bp of the 3' ends of five other elements were compared to IS711 and to each other, minor nt sequence variations were found amongst most of them. Similar to several other transposable elements, IS711 has overlapping ORFs rather than one long ORF extending the length of the element. Even though only ten B. ovis IS711 elements were characterized, in three cases we found these elements flanked by either identical or similar nt sequences. This suggests that some target sites are hot spots for insertion and that some of the elements may be duplicated by mechanisms other than transposition. No DNA or protein database entries had an obvious resemblance to either IS711 or its deduced gene products. PMID- 8224886 TI - Cloning and sequence analysis of the gene encoding the crystalline surface layer protein of Rickettsia typhi. AB - The nucleotide sequence of the gene (slpT) encoding the crystalline surface layer protein (SLP) of Rickettsia typhi was determined. The slpT gene consists of 4935 bp coding for a 1645-amino-acid (aa) protein containing a predicted signal peptide at the N terminus. The size of the predicted SLP exceeds the observed size (135 kDa) on SDS-PAGE. The N-terminal aa sequence of the 32-kDa protein of R. typhi reported by Hackstadt et al. [Infect. Immun. 60 (1992) 159-165] was found in the C-terminal portion of the deduced aa sequence, suggesting that the product of slpT is processed into the mature SLP and the 32-kDa protein. PMID- 8224887 TI - Isolation and characterization of the Rickettsia prowazekii gene encoding the flavoprotein subunit of succinate dehydrogenase. AB - The gene (sdhA) coding for the flavoprotein subunit (SdhA) of succinate dehydrogenase of the obligate intracellular parasitic bacterium, Rickettsia prowazekii, has been isolated using an oligodeoxyribonucleotide probe to the conserved flavin adenine dinucleotide (FAD)-binding region of characterized flavoproteins. Nucleotide (nt) sequence analysis revealed an open reading frame (ORF) of 1791 bp capable of encoding a protein of 596 amino acids (aa) with a deduced M(r) of 65,444. The deduced aa sequence, when compared to the flavoprotein subunits of Escherichia coli, Bacillus subtilis, Saccharomyces cerevisiae and Bos taurus, revealed 52.8, 34.0, 65.8 and 52.0% aa identity, respectively. R. prowazekii SdhA produced in E. coli minicells and analyzed by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis (SDS-PAGE) migrated as a protein of approximately 63 kDa, comparable to the size of the deduced protein. In addition, two proteins of approximately 12 and 41 kDa were also produced in the E. coli minicells. The production of these proteins resulted from additional translational starts within the SdhA coding sequence, suggesting differences between the translational start signals of E. coli and R. prowazekii. Despite the similarity of R. prowazekii SdhA to that of E. coli, the R. prowazekii SdhA did not complement an E. coli sdhA mutant. In addition, analysis of the nt sequence immediately upstream from R. prowazekii sdhA revealed that the rickettsial sdh gene organization differs from that of E. coli and B. subtilis. PMID- 8224888 TI - A strain-specific cyclin homolog in the fungal phytopathogen Colletotrichum gloeosporioides. AB - The fungus, Colletotrichum gloeosporioides, which infects the tropical pasture legume, Stylosanthes guianensis, contains highly variable mini-chromosomes. The transcription of strain-specific genomic DNA clones previously isolated from one variable mini-chromosome was investigated by using these clones to screen a cDNA library prepared from the fungus grown in liquid medium. A cDNA clone was obtained with one of the genomic clones and was sequenced. A single long open reading frame of 259 amino acids (aa) was detected with significant homology to cyclin proteins in other organisms. Northern blot analysis indicated that the cDNA corresponded to a low-abundance mRNA (approximately 0.001% of poly(A)+RNA). Southern blot analysis indicated that genes encoding this mRNA were discontinuously distributed in this fungal species, indicating it encodes a dispensable function. This result suggests that natural populations of fungi may have variable complements of cyclin-encoding genes. PMID- 8224889 TI - Isolation and complete sequence of the purL gene encoding FGAM synthase II in Lactobacillus casei. PMID- 8224890 TI - Genome rearrangements by residual IS10 elements in strains of Escherichia coli K 12 which had undergone Tn10 mutagenesis and fusaric acid selection. AB - Mutant strains selected as survivors of the lethal overexpression of a plasmid encoded bovine somatotropin-beta-galactosidase fusion protein were found to include instances where an IS10 element had transposed from the chromosome into the fusion protein structural gene on the plasmid. Two distinct types of IS10 elements were found in these mutants, the well-known IS10R and a novel hybrid element composed of portions of both IS10R and IS10L. The strain in which the selection scheme was carried out had been constructed in a series of steps, including alteration of two loci by Tn10-mediated intramolecular transposition involving fusaric acid (FA) selection for loss of tetracycline resistance. Genetic dissection of this strain revealed that one of these altered loci was an origin for both types of IS10 elements, while the other locus was an origin for only IS10R elements. The finding that residual IS10 elements, left after FA selection for Tcs derivatives of Tn10-containing strains, can be a significant source of spontaneous mutation should be of interest to workers using strains that have been 'cured' of Tn10 in this way. PMID- 8224891 TI - Construction and use of a new broad-host-range lacZ transcriptional fusion vector, pHRP309, for gram- bacteria. AB - A new lacZ transcriptional fusion vector, pHRP309, based on the IncQ plasmid RSF1010, was constructed and shown to be easily mobilized into a variety of Gram- eubacteria. We also developed a two-step cloning procedure to facilitate the cloning of small promoter fragments into the fusion vector. A set of 'cohort' vectors was constructed which allowed directed cloning of fragments downstream from an omega streptomycin/spectinomycin-resistance cassette while maintaining multiple flanking restriction sites. The omega cassette provides a selectable antibiotic-resistance marker for cloning promoters into the fusion vector and makes mapping to determine fragment orientation unnecessary. The presence of the omega cassette also decreases background beta-galactosidase activity by decreasing readthrough transcription from plasmid sequences. The fusion vector carries a gentamicin-resistance-encoding gene as the selectable marker and can therefore be used in Tn5 (kanamycin-resistant) and Tn10 (tetracycline-resistant) mutant strains. Since pHRP309 is a member of the IncQ incompatibility group, it is compatible with IncP cloning vectors and can be used in strains carrying cloned regulatory genes. Using this system, we cloned the positively regulated Pseudomonas putida pcaI promoter and studied its regulation. PMID- 8224892 TI - Characterization of the genes controlling the biosynthesis of the polyketide phytotoxin coronatine including conjugation between coronafacic and coronamic acid. AB - Pseudomonas syringae pv. glycinea PG4180 produces a chlorosis-inducing phytotoxin, coronatine (COR), which consists of a polyketide component, coronafacic acid (CFA), which is coupled via amide bond formation to coronamic acid (CMA), an ethylcyelopropyl amino acid (aa) derived from isoleucine. P. syringae pv. syringae strains PS51 and PS61, which do not synthesize coronafacoyl compounds (conjugates between CFA and aa), acquired the ability to produce CFA and COR when transformed with p4180A, a 90-kb indigenous plasmid in PG4180. Tn5 mutagenesis indicated that the COR biosynthetic genes in PG4180 are clustered within a 30-kb region on p4180A. The phenotype of selected COR-defective mutants was determined by supplying them with CFA and CMA and by complementation studies with cloned DNA from the COR biosynthetic cluster. Using this approach, the regions encoding CFA and CMA synthesis and coupling activity were localized to the 24-, 12.5- and 2.3-kb regions of the cluster, respectively. Mutants in a 6-kb region required the addition of both CFA and CMA for COR synthesis, which may indicate a regulatory role for this part of the cluster. PS51 and PS61 transconjugants containing cloned DNA from the coupling region produced COR when supplied with CFA and CMA, indicating that coupling activity was cloned and expressed in bacteria lacking the COR biosynthetic cluster. PMID- 8224893 TI - An efficient expression and secretion system based on Bacillus subtilis phage phi 105 and its use for the production of B. cereus beta-lactamase I. AB - A novel expression system based on the Bacillus subtilis bacteriophage phi 105 has been developed to permit the high-level synthesis and secretion of beta lactamase I (BlaI) from Bacillus cereus. Shotgun insertion of a promoterless lacZ gene into the phage genome permitted the identification of a clone producing large amounts of beta-galactosidase (beta Gal), indicating the transcription of the reporter gene from a strong phage promoter. The insertion also blocked lysis of the host cell. Although the insertion in the original prophage was complex, plasmid vectors and prophage derivatives have been developed to facilitate the replacement of lacZ with other genes for expression. The new prophages contain two additional mutations: an ind mutation, which greatly enhances the normally poor transformability of phi 105 lysogens, and a cts mutation, which allows thermo-induction of phage development and protein production. Induction of a derivative prophage containing the blaI gene from B. cereus resulted in the production of up to 500 micrograms of secreted BlaI per ml of culture supernatant. PMID- 8224894 TI - The cloning and sequencing of the genes encoding phytase (phy) and pH 2.5-optimum acid phosphatase (aph) from Aspergillus niger var. awamori. AB - The genes encoding phytase (EC 3.1.3.8) and pH 2.5-optimum acid phosphatase (EC 3.1.3.2) have been cloned and sequenced from Aspergillus niger var. awamori. The translated nucleotide sequences yielded polypeptides of 467 and 479 amino acids (aa) for phytase and acid phosphatase, respectively. The genes were isolated using oligodeoxyribonucleotide probes based on the aa sequences of the purified proteins. Recombinant A. niger var. awamori strains carrying additional copies of the gene sequences demonstrated elevated enzyme activities. PMID- 8224895 TI - Expression and antisense inhibition of transgenes in Phytophthora infestans is modulated by choice of promoter and position effects. AB - Procedures were identified for manipulating the expression of genes in the oomycete fungus, Phytophthora infestans. The activities of five putative promoter sequences, derived from the 5' regions of oomycete genes, were measured in transient assays performed in protoplasts and in stable transformants. The sequences tested were from the ham34 and hsp70 genes of Bremia lactucae, the actin-encoding genes of P. infestans and P. megasperma, and a polyubiquitin encoding gene of P. infestans. Experiments using the GUS reporter gene (encoding beta-glucuronidase) demonstrated that each 5' fragment had promoter activity, but that their activities varied over a greater than tenfold range. Major variation was revealed in the level of transgene expression in individual transformants containing the same promoter::GUS or promoter::lacZ fusion. The level of expression was not simply related to the number of genes present, suggesting that position effects were also influencing expression. Fusions between the ham34 promoter, and full-length and partial GUS genes in the antisense orientation blocked the expression of GUS in protoplasts and in stable transformants. PMID- 8224896 TI - Structures of the Euplotes crassus Tec1 and Tec2 elements: identification of putative transposase coding regions. AB - The Tec1 and Tec2 transposon-like element families of Euplotes crassus are highly unusual in that all 30,000 copies of each family are excised from the genome during a discrete time period of macronuclear development. Complete nucleotide sequences were generated for the Tec1-1 and Tec2-1 elements, representing the Tec1 and Tec2 families. Open reading frames (ORFs) are conserved in position and sequence between the two elements, although sequences that comprise one ORF (ORF2) of Tec1-1 are split into two overlapping ORFs (ORFs 2A and 2B) in Tec2-1. ORF1 in Tec1-1, its homolog in Tec2-1 and one of the overlapping ORFs from Tec2-1 (ORF2B) contain TGA codons, which may be translated as Cys, as observed for two other Euplotid genes. Sequence analyses of ORFs from other members of each element family indicate that the families are distinct from each other and are highly conserved within each family. Computer searches of sequence databases have revealed sequence similarity between Tec ORF1s and the previously described Tc1 IS630 family of transposases which includes ORFs from bacterial, nematode and insect transposons. PMID- 8224897 TI - C-terminal deletion mutants of the FokI restriction endonuclease. AB - We have constructed two C-terminal deletion mutants of the FokI restriction endonuclease by using the polymerase-chain-reaction technique and expressed them in Escherichia coli. The two mutant proteins (MP) of 41 and 30 kDa, were purified to homogeneity and their DNA-binding properties were characterized. The 41-kDa MP specifically binds the DNA sequence, 5'-GGATG/3'-CCTAC, like the wild-type (wt) FokI, but does not cleave DNA. The 30-kDa MP does not bind DNA. The affinity of the 41-kDa MP for the DNA substrate is comparable to that of wt FokI. The 41-kDa MP interacts with its substrate like the wt FokI, as revealed by hydroxyl radical footprinting experiments. In the presence of a DNA substrate, the 41-kDa MP is cleaved by trypsin into a 30-kDa N-terminal fragment and an 11-kDa C-terminal fragment. Addition of the HPLC-purified 11-kDa C-terminal fragment to the 30-kDa MP restores its sequence-specific DNA-binding property. These results confirm that the N-terminal 41-kDa fragment of the FokI ENase constitutes the DNA recognition domain of the ENase. PMID- 8224898 TI - XcmI as a universal restriction enzyme for single-stranded DNA. AB - Single-stranded DNA can be cleaved into defined fragments at any predetermined site by interaction with a specially designed oligodeoxyribonucleotide (oligo) adaptor and the class-IIN restriction endonuclease, XcmI. The oligo adaptor has the structure [sequence: see text]. Upon hybridization to the target DNA through the central 9-nucleotide region and with the addition of XcmI, the template DNA is specifically cleaved to near completion. Hairpin structures on the template close to the hybridization site reduce the efficacy of cleavage. PMID- 8224899 TI - Synthesis of two bacteriophage lambda S proteins in an in vivo system. AB - Bacteriophage lambda has two genes which are essential for lysis: R, a gene encoding a 158-amino-acid (aa) transglycosylase that attacks the peptidoglycan, and S, a gene encoding two inner-membrane-associating proteins, designated S105 and S107 for their predicted lengths in aa residues. S105 and S107 are thought to have opposing roles in lysis, with the former acting as the lethal lysis effector and the latter as a lysis inhibitor. Here, we used a T7-polymerase-mediated expression system to show that S105 and S107 are synthesized at a constant ratio of about 2.5:1 throughout the period leading up to lysis, indicating that lysis scheduling does not require a translationally controlled switch from inhibitor (S107) to effector (S105) synthesis. However, evidence is presented that the mRNA sequences immediately 5' to the ribosome-binding site (RBS) of the S gene are required for the rather limited translation, but not the stability, of the S mRNA. No difference could be found in the pattern of ternary complex formation over the two S start codons in in vitro toe-printing assays with the wild-type mRNA and with mRNA deleted of the upstream sequences. Nevertheless, these results may suggest a role for translational control in S gene expression, if not in its temporal regulation or in the partition between S105 and S107 production. PMID- 8224900 TI - Cloning, purification and characterization of the BseCI DNA methyltransferase from Bacillus stearothermophilus. AB - The gene (bseCIM) encoding the BseCI DNA methyltransferase (MTase; M.BseCI) from a Bacillus stearothermophilus species was cloned and expressed in Escherichia coli using plasmid vector pBR322. Selection of transformants carrying bseCIM was based on the resistance of the modified plasmid to cleavage by BseCI. The MTase was purified to homogeneity and further characterized. Its size as determined by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis and size exclusion chromatography was 68 kDa, suggesting that the MTase exists as a monomer. When phage lambda DNA was used as a substrate, the optimum temperature for MTase activity was determined to be 50-55 degrees C and optimum pH approx. 7.4. M.BseCI is inhibited by concentrations of NaCl and KCl greater than 50 mM, and it does not require Mg2+ for activity. Finally, M.BseCI methylates the 3' adenine residue in the sequence, 5'-ATCGAT-3', similarly to its isoschizomer M.ClaI. PMID- 8224901 TI - A novel phage lambda replacement Cre-lox vector that has automatic subcloning capabilities. AB - We have developed a novel phage lambda replacement cloning vector, lambda pAn. lambda pAn allows one to automatically subclone the insert as a plasmid using the Cre-loxP site-specific recombination system. This eliminates the need to subclone insert fragments and permits the rapid structural analysis of insert DNA. lambda pAn is similar to other phage lambda replacement vectors taking inserts ranging in size from 5 to 19 kb. We have placed the pyrG gene of Aspergillus nidulans on the vector as a nutritional selective marker for transformation. We have developed this vector as part of an overall plan to facilitate the cloning of dominant extragenic suppressor mutations from A. nidulans, but also know that it is a generally useful vector for the purposes of isolating genomic clones without the need to subclone from the phage lambda vector. PMID- 8224902 TI - New improved lacZ gene fusion vectors. AB - New plasmid vectors suitable for creating fusions with the lacZ gene have been developed. These vectors represent an improvement over currently available vectors and possess the following features: (1) an undetectable background beta galactosidase (beta Gal) activity in the absence of fusion, (2) an extended multiple cloning site (MCS), and (3) the ability to conveniently subclone in any one of three translational frames. Medium- and high-copy-number versions of these vectors have been developed. PMID- 8224903 TI - Analysis of c-Myc domains involved in stimulating SV40 replication. AB - We have demonstrated previously that overproduction of c-Myc, N-Myc and, to a lesser extent, L-Myc facilitates the replication of simian virus 40 (SV40)-based vectors in human lymphoid cells. Using a series of c-myc deletion mutants, we investigated which c-Myc regions are important in stimulating SV40 replication. The ability of c-Myc to promote SV40 replication was significantly reduced by deletions in the second exon domain, formerly shown to be crucial for c-Myc's transforming capacity. The c-myc mutants with a disrupted basic region (b) or leucine zipper (Zip) motif were also unable to stimulate SV40 replication. These regions are implicated in protein-DNA and protein-protein interactions, respectively, suggesting that the c-Myc protein might be associated with the DNA protein replication complex. We present data obtained from gel mobility shift assays and from an immunocomplex-binding assay substantiating this hypothesis. PMID- 8224904 TI - Epoc-1: a POU-domain gene expressed in murine epidermal basal cells and thymic stromal cells. AB - POU-domain transcription factors are known as developmental regulators which control organ development and cell phenotypes. In order to clarify the roles of POU-domain transcription factors in cell differentiation, we cloned a novel POU family gene, Epoc-1, from a murine thymus cDNA library. The amino acid (aa) sequence of the POU-specific domain of Epoc-1 is almost identical to those of Oct 1 and Oct-2. However, within the POU-homeodomain, 13 out of 60 aa differ between Epoc-1 and Oct-2. Recombinant Epoc-1 products were found to bind specifically to the octamer sequence. Epoc-1 was found to be expressed in skin, thymus, stomach and testis. In situ hybridization experiments and RNase protection assays indicated that Epoc-1 is expressed in the epidermal basal cells of the skin, which contain stem cells unipotent for keratinocyte differentiation and in thymic stromal elements. These results suggest that Epoc-1 might be one of the developmental regulators which controls epidermal development and thymic organogenesis. PMID- 8224905 TI - Murine ecto-5'-nucleotidase (CD73): cDNA cloning and tissue distribution. AB - The murine cDNA, encoding the purine catabolic enzyme, ecto-5'-nucleotidase (NT), was cloned and the tissue-specific distribution of both the mRNA and enzyme activity was examined. Starting with kidney RNA and primers based on the known rat sequence, reverse transcriptase-polymerase chain reaction (RT-PCR) was utilized to obtain the complete sequence for the translated portion of the murine cDNA. Murine NT is 94% identical to human NT at the amino acid (aa) level and 86% identical at the nucleotide (nt) level. NT enzyme assays revealed greater than tenfold more NT activity in mature vs. immature murine T- and B-lymphocytes. A similar increase in NT activity was also found when the pre-B-cell line, 70Z/3, was induced to produce surface kappa light chains with lipopolysaccharide (LPS) and gamma-interferon (gamma-IFN). Thus, culture systems in which murine lymphocytes mature may be useful for examining the mechanisms of NT gene regulation, as well as the function of NT in the immune system. In tissues, enzyme activity varied over 30-fold, from the lowest levels in skeletal muscle, thymus and spleen to highest in placenta, kidney and forestomach. Levels of mRNA, as determined by RNase protection assay, showed increased NT expression in the early gestation site, as compared to non-pregnant uterus, and in day-19.5 placenta, as compared to day-13 chorioallantoic placenta. Messenger RNA levels were in general proportional to enzyme activity, except in the lung and glandular stomach where mRNA levels were higher than expected, based on enzyme activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224906 TI - The sequence of the mouse phosducin-encoding gene and its 5'-flanking region. AB - Phosducin (Pd), a principal protein of retinal photoreceptor cells, modulates the phototransduction cascade by interacting with transducin. A recent report indicated that Pd and the G-protein-inhibitor protein (GIP) in brain are virtually identical. Here, we have sequenced the complete mouse clone (P1-AT) carrying the Pd gene, 3026 bp of its 5'-flanking region, and cDNAs generated from the retinal mRNAs. Gene Pd is 15 kb in length and has four exons. The splice sites for donor and acceptor were in good agreement with the GT/AG rule. Deduced Pd amino acid sequences were highly homologous to those of human, bovine and rat. In addition, we found more than one similar Pd gene and two different mRNAs. The P1-AT clone encodes one of the Pd mRNAs. These results open the possibility that the multiple Pd genes encode multiple Pd. PMID- 8224907 TI - Characterization of rat pseudogenes for enhancer factor I subunit A: ripping provides clues to the evolution of the EFIA/dbpB/YB-1 multigene family. AB - Genomic Southern blot analysis of rat EFIA (gene encoding enhancer factor I subunit A) reveals a complex band pattern when cDNA subfragment probes are used. Screening a rat genomic library with a rat EFIA cDNA probe yields two different processed EFIA pseudogenes, designated rat psi EFIA#(2/3) and #(4/7), in addition to two other different, but less extensively characterized clones. psi EFIA#(4/7) has no open reading frame (ORF) sequences. psi EFIA#(2/3) contains two ORFs (83 and 178 codons), the products of which (if expressed) might be negative-acting EFIA transcription factors. Located nearly 0.6 kb upstream from psi EFIA#(2/3) is a perfect 69-bp dinucleotide (CT) tandem repeat, a sequence element associated with other isolated pseudogenes. Additionally, the 3' end of this processed gene is interrupted by an unusual retroposon, an inverted dimeric B1-like short interspersed repetitive element (SINE). The isolation of several independent clones of the same EFIA processed pseudogenes indicates that they comprise a significant component of the rat EFIA copy multiplicity. The phenomenon of repeat induced point mutagenesis (ripping) at rat EFIA pseudogene CpG doublets occurs at a frequency at least 6.5 times higher than predicted from random mutagenesis. This is consonant with the proposal that ripping may be the mechanism which inactivates the ectopic recombination potential of the rat EFIA pseudogenes. PMID- 8224908 TI - Identification of conserved potential regulatory sequences of the protamine encoding P1 genes from ten different mammals. AB - In order to detect regulatory conserved DNA elements within the protamine 1 encoding gene (P1) promoter, we have sequenced this region from the rat, guinea pig, gorilla, orangutan, anubis baboon and red monkey P1 genes and compared it to the homologous human, bull, boar and mouse nucleotide (nt) sequences. We demonstrate the presence of a consensus sequence, HSMCYTCAYAAT (Prot1C: protamine 1 consensus), from nt position -64 to position -53 in all P1 genes whose promoter sequences are now known. We also show that sequences similar to Prot1C are found in the promoter region of other testis-specific genes, such as the transition protein 1-encoding gene promoter which is thought to have derived from the P1 genes. The relevance of this conserved element in the expression of P1 genes is strongly supported by the recent demonstration of a mouse testis trans-acting factor [Tet-1; Tamura et al., J. Biol. Chem. 267 (1992) 4327-4332] which binds and matches in the mouse the first 11 bp of the corresponding consensus Prot1C sequence reported here. Another highly conserved element (TGTGAGG) has been identified 20 +/- 3 nt upstream from Prot1C. This sequence forms a perfect palindrome with the central 7 nt of Prot1C and is absent in the homologous region of other genes. Further upstream, at positions -113 to -132, a third highly conserved region is present (MATGCCCATATWTGGRCAYG) which is similar to the c-fos SRE (serum-response element) and contains the central core common to all SREs. This element has not been found in the homologous region of other sperm-specific genes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224909 TI - One-step purification and characterization of a lignin-specific O methyltransferase from poplar. AB - O-Methyltransferases (OMT; EC 2.1.1.6) play an important role in the synthesis of lignin precursors by catalyzing the O-methylation of o-diphenolic substrates such as caffeic acid (CA) and 5-hydroxyferulic acid (5OH). Here, we report on the purification of a lignin-specific OMT (38 kDa) from poplar (Populus trichocarpa x P. deltoides). The OMT was purified from xylem by a single affinity chromatography step on adenosine agarose. The enzyme uses both CA and 5OH as substrates. We previously have reported the cloning of a corresponding OMT cDNA [Dumas et al., Plant Physiol. 98 (1992) 796-797]. Expression of this OMT cDNA in Escherichia coli further confirmed the identity of the clone. Genomic hybridization demonstrates the presence of one or two OMT genes per haploid poplar genome. RNA gel blot hybridization shows high levels of steady-state OMT mRNA in the xylem of young poplar trees, as compared to the levels in leaves. PMID- 8224910 TI - Translational properties of the human papillomavirus type-6 L1-coding mRNA. AB - A cDNA encoding a bicistronic mRNA, E1E4L1, which was generated by double splicing of the E1, E4 and L1 genes, of the type-6 human papillomavirus (HPV-6), was cloned. The E1E4 and L1 open reading frames (ORFs) in this cDNA were expressed in COS-1 or CV-1 cells as fusion proteins with Escherichia coli beta galactosidase (beta Gal), and the products were analyzed by immunoprecipitation and enzyme assay. The results showed that the translational efficiency of the L1 ORF was about 9-15-fold less efficient than that of the E1E4 ORF. Substitution of the ATG of the E1E4 ORF with AAG increased translation of the L1 ORF about 30 fold. Lengthening of the intercistronic sequence to 31 bp, equivalent in length to the bicistronic HPV-1 mRNA, showed little translational effect relative to the wild-type 12-bp intercistronic sequence. (Carets [] represent splicing of RNA.) PMID- 8224911 TI - Cloning a pseudogene and cDNA encoding a 17-kDa ribosomal protein from mouse: structure and regulation of expression. AB - An rp lambda 5 cDNA encoding a ribosomal protein (r-protein) and a pseudogenic form of the corresponding gene (rp lambda 7) have been cloned from mouse. This cDNA codes for a highly basic protein of 160 amino acids (aa) with a deduced M(r) of 17,601, and most likely represents the species homolog of a recently cloned rat cDNA, which has been proposed to encode a homolog of the yeast r-protein, YL43. The entire rp lambda 5 gene encompasses less than 1.5 kb of genomic DNA and apparently is composed of only two exons, as deduced from sequence comparison with its very similar pseudogenic variant, rp lambda 7. Southern analysis, using the rp lambda 5 cDNA as a probe, indicates the existence of a great number of highly related sequences in the mouse genome. The mRNA for rp lambda 5 is approximately 800 nucleotides (nt) long and is found to be ubiquitously expressed at high levels in embryonic and adult mouse tissues, as shown by Northern and in situ analyses. Retinoic acid (RA) seems to have a moderate down-regulatory effect on this mRNA in differentiating P19 embryonal carcinoma cells. Several degenerate/nondegenerate RA-response element (RARE) motifs are found within 560 bp upstream from the degenerate start codon in rp lambda 7. However, it is unknown whether this RA effect is exerted at the transcriptional and/or posttranscriptional levels. PMID- 8224912 TI - Structural features of the 5' upstream regulatory region of the gene encoding rat amyloid precursor protein. AB - The 5' upstream regulatory region of the gene encoding the rat amyloid precursor protein (APP) was cloned and sequenced. It lacks both a TATA box and a CAAT box, has a high G + C content (68%), is 89% homologous to the corresponding region of the mouse APP gene, and 82% homologous to the corresponding region of the human APP gene. This region contains putative regulatory elements both 5' and 3' to the probable transcription start point (tsp). There are consensus DNA sites for the binding of SP1, AP2, AP4 and GC factor (GCF) proteins, and two GC boxes with the consensus sequence, 5'-GGGYGCRG. Potential regulatory sites with only a single mismatch to the consensus sequences include three SP1, one AP1, five AP2, and two GCF sites, as well as one GC box. There are also six potential stem-loop secondary structures (SSS) near the probable tsp. A consecutive series of elements, consisting of a GC box, AP2 site, three SSS, two SP1 sites, and AP4, AP1 and GCF sites just upstream from the probable tsp, are well-conserved between the rat, mouse and human sequences. An additional AP2 site, two GC boxes, and two additional SSS appear to be conserved between species. However, two possible rat SP1 sites, three possible rat AP2 sites, and two possible rat GCF sites are lacking in the human. On the other hand, the rat sequence is missing four potential SP1 sites, four potential AP2 sites, and nine potential GC boxes which are found in the human sequence.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8224913 TI - Isolation and expression of a rat liver cDNA encoding phosphoglucomutase. AB - A cDNA encoding phosphoglucomutase (PGM) has been isolated from a rat liver cDNA library following screening with a polymerase chain reaction product. The cDNA was found to contain a 53-base-pair (bp) 5' untranslated region (5' UTR), a single start codon and consensus initiation sequence, an open reading frame (ORF) of 1686 bp, and a 3' untranslated tail. A comparison to the rabbit and human muscle PGM cDNAs [Whitehouse et al., Proc. Natl. Acad. Sci. USA 89 (1992) 411 415] showed 90% identity of rat cDNA to both, while a comparison to the deduced amino acid sequences showed 97 and 96% identity, respectively. Northern blot analyses determined that PGM was encoded by a single mRNA in rat liver, of approximately 2.2 kb. Following transfection of COS-7 cells with a plasmid containing the entire PGM ORF, indirect immunofluorescence analyses using a PGM specific monoclonal antibody determined that approximately 5% of the cells displayed 50-100 times greater fluorescence than that seen in the remainder of the cells or in mock transfects. The enhanced production of PGM was also demonstrated by Western blotting and by enzymatic activity assays. PMID- 8224914 TI - Isolation and analysis of the rat genomic sequence encoding Cu/Zn superoxide dismutase. AB - A rat genomic DNA (SOD1) encoding Cu/Zn superoxide dismutase (SOD1) (superoxide; superoxide oxidoreductase, EC 1.15.1.1) was cloned and sequenced. The rat SOD1 gene consisted of five exons and four introns spanning about 6 kb. The transcription start point (tsp) was observed 93 bp upstream from the ATG codon by primer extension analysis. The 5'-flanking sequence of SOD1 contained two CCAAT box motifs, a TATA box and four GC-like boxes. In the 3'-flanking region of SOD1, a polyadenylation signal, consensus sequence YGTGTTYY, and a G/T cluster were observed. A rat identifier (ID) sequence, a repetitive element of the rat genome, was located at between 569 and 484 bp upstream from the tsp. PMID- 8224915 TI - Cloning of the cDNA encoding human xanthine dehydrogenase (oxidase): structural analysis of the protein and chromosomal location of the gene. AB - The primary structure of human xanthine dehydrogenase (hXDH) was determined by cloning and sequence analysis of the cDNAs encoding the enzyme. The nucleotide (nt) sequence has an open reading frame of 3999 nt encoding a protein of 1333 amino acids (aa) with a calculated M(r) of 146,604. The deduced aa sequence of hXDH is homologous to the previously reported rat XDH (rXDH) and Drosophila melanogaster XDH sequences with identities of 90.2 and 52.0%, respectively. The aa residues involved in both the reversible and the irreversible conversion from the dehydrogenase type to the oxidase type of rXDH are completely conserved between the rat and the human enzymes. This implies that the molecular mechanisms of the conversion of hXDH from dehydrogenase to oxidase are common to those of the well-characterized rXDH. Five sequence variations were detected in the isolated cDNA clones. Spot blot hybridization using flow-sorted human chromosome revealed that the hXDH-encoding gene (hXDH) was located on chromosome 2. PMID- 8224916 TI - Sequence and organization of the human N-formyl peptide receptor-encoding gene. AB - The human FPR1 gene encodes the N-formyl peptide receptor, a G protein-coupled receptor (GPCR) that mediates the activation of mature myeloid cells by bacterial N-formyl oligopeptides. To investigate the molecular basis for myeloid-specific production of this receptor, we have cloned and sequenced FPR1. The gene is organized into three exons and two introns that span 6 kb. The coding block lacks introns. Yet, the transcription start point (tsp) is separated from the start codon by 4902 bp consisting of three exons and two large introns. Two mRNAs are produced by alternative splicing of exon 2 in HL-60 neutrophils and normal blood monocytes. The region 5' to the tsp contains three pyrimidine-rich segments, a feature that has been observed in other myeloid-specific genes. One complete Alu repeat is found in each intron and in the 3'-flanking region 317 bp downstream of the polyadenylation signal. Thus, FPR1 is a small myeloid-specific gene that is expressed as two alternatively spliced mRNAs encoding the same protein. PMID- 8224917 TI - Production of human plasma retinol-binding protein in Escherichia coli. AB - We designed a polymerase chain reaction (PCR) primer pair which allowed us to clone the cDNA coding for the human plasma retinol-binding protein (hRBP) into an Escherichia coli expression vector. Production of hRBP was confirmed by probing Western blots with antisera against plasma hRBP. Purification and characterization of the E. coli-produced plasma hRBP are also described. The availability of this expression system makes it possible to obtain large quantities of hRBP to facilitate our continuing studies of retinol and RBP metabolism. PMID- 8224918 TI - Sequence of the human lens beta B2-crystallin-encoding cDNA. AB - This study reports the nucleotide (nt) sequence of the human beta B2-crystallin (h beta B2-Cry)-encoding cDNA (h beta B2-cry). The h beta B2-cry gene encodes a major structural protein in the lens of the vertebrate eye. Sequence information obtained from mouse genomic clones and the mouse beta B2-cry cDNA (m beta B2-cry) sequence was used to design oligodeoxyribonucleotide primers, corresponding to exon 1, transcription start points, and termination and polyadenylation sites, that were used in the polymerase chain reaction (PCR) to generate full-length cDNA clones from total human lens RNA. In addition, cDNA libraries were made with lambda gt11 from both human fetal and adult lens tissue RNAs, and screened with a m beta B2-cry cDNA clone. The full-length h beta B2-cry cDNA is 721 bp and contains an open reading frame of 612 nt. It codes for a 23-kDa protein of 205 amino acid residues. Comparison of the overall nt and deduced aa sequences shows a greater similarity of h beta B2-cry to bovine beta B2-cry than to m beta B2-cry or rat beta B2-cry. PMID- 8224919 TI - Cloning and sequencing of 5' flanking sequence from the gene encoding 2S storage protein, from two Brassica species. AB - Using oligodeoxyribonucleotide primers and the polymerase chain reaction, we have cloned and sequenced about 1.2 kb of upstream sequences from two members of the 2S seed storage protein-encoding gene family from Brassica juncea and B. oleracea. The two sequences bear more than 90% homology and have characteristic seed-specific promoter motifs. The high degree of sequence conservation indicates that this napin-encoding gene family evolved earlier than the divergence of the three primary Brassica species and their amphidiploids, and the sequences have been conserved due to some metabolic constraints in seed development. PMID- 8224920 TI - Sequence of the cDNA encoding an actin homolog in the crayfish Procambarus clarkii. AB - A cDNA library was constructed by using mRNAs purified from crayfish (Procambarus clarkii) muscle. Using a homology search of the nucleotide (nt) sequences, a clone of the library was found to encode a protein homologous to actin (Act). The insert fragment of this cDNA clone was 1072 nt in length. The amino acid sequence deduced from the nt sequence showed significant similarity to Act of various organisms as follows: 88.1% to Drosophila melanogaster, 88.2% to silk worm, 87.3% to brine shrimp, 86.3% to rat, and 86.3% to human (% identity). PMID- 8224921 TI - Two distinct cDNA sequences encoding the human helix-loop-helix protein Id2. AB - Two distinct Id2 cDNA clones, Id2A and Id2B (inhibitor of differentiation), have been isolated from a human heart cDNA library and sequenced. The deduced amino acid sequences show nearly complete identity, except for the nonsense mutation at codon 37 in Id2B. Characterization of the genomic clones revealed that Id2B is encoded by an intronless gene which probably arose after the evolutionary divergence of rats and humans. PMID- 8224922 TI - Human and baboon integrin beta 5 subunit-encoding mRNAs have alternative polyadenylation sites. AB - A cDNA probe, encoding part of the human integrin polypeptide, GPIIIa (beta 3), was used to screen a cDNA library derived from baboon smooth muscle polyadenylated mRNA. One cross-hybridizing clone, lacking the baboon equivalent of 478 bp of the human sequence, was found to be 98% similar to a human cDNA encoding the beta 5-chain of the receptor for vitronectin, a cell adhesion molecule. The baboon mRNA terminates at 3' position 212 nucleotides upstream from the polyadenylation site of the aligned human mRNA. PMID- 8224923 TI - Dementia: making diagnosis less confusing. PMID- 8224924 TI - Osteoarthritis: is it inflammatory or not? PMID- 8224925 TI - Vascular dementia: stroke prevention takes on new urgency. AB - Vascular dementia is a clinical syndrome of acquired intellectual impairment resulting from brain injury due to a cerebrovascular disorder. It is a complex diagnosis, and diagnostic criteria vary. In community practice, the physician can probably make the diagnosis based on the history and medical examination. CT demonstration of one or more infarcts increases the likelihood of this diagnosis. Hypertension is a major risk factor for vascular dementia. Others include smoking, hyperlipidemia, atrial fibrillation, diabetes, and a sedentary lifestyle. Cerebrovascular disease is an important cause of cognitive decline in older patients. Therefore, it is important to recognize risk factors for stroke and institute measures for prevention. PMID- 8224926 TI - Mild heart failure: why the switch to ACE inhibitors? AB - Management of congestive heart failure in the past has focused on sodium and fluid restriction, rest, and digitalis glycosides. Now, significant new evidence justifies early and aggressive ACE inhibitor therapy in patients with asymptomatic or mildly symptomatic LV dysfunction. ACE inhibitors reduce the likelihood of symptomatic heart failure in asymptomatic patients with reduced ejection fraction. Patients with reduced LV function following acute MI who receive ACE inhibitors have a decreased risk of death, a lower probability of developing systematic heart failure, and fewer MI recurrences. Hypotension and azotemia can be avoided by reducing the concomitant dose of diuretics and carefully titrating the ACE inhibitor dosage to target levels. PMID- 8224927 TI - Exercise for older patients: why it's worth your effort. AB - Two-thirds of all adults age 65 or older are either irregularly active or completely sedentary. With this inactivity comes an increased risk of chronic diseases, including coronary heart disease, hypertension, diabetes, osteoporosis, and depression. Adequate aerobic exercise--even when started as late as age 60- is associated with a 1- to 2-year increase in life expectancy, as well as increased functional independence. Even chairbound patients can benefit from a program of simple exercises. To help prevent injuries, your exercise prescription should include stretching exercises and exercises to strengthen the muscles surrounding weak joints. PMID- 8224928 TI - Health system reform: the sudden shift toward primary care. PMID- 8224929 TI - Stargardt's disease/fundus flavimaculatus: psychophysical and electrophysiologic results. AB - Retinal functions were analyzed psychophysically and electrophysiologically in 73 patients (146 eyes) with Stargardt's disease/fundus flavimaculatus. Patients were classified into types 1, 2, 3, and 4; patients with type 3 were subdivided into 3E and 3L (early and late onset of initial symptoms). Most had visual acuity (VA) of 20/200 or greater at initial testing. VA declined 0.25 octave/year during follow-up (mean, 6.1 years). Four of 16 patients (25%) older than 40 years had VA of 20/200 or less in the stronger eye. Psychophysical tests (flicker profile, central scotoma, dark adaptation) showed variable degrees of abnormalities. Electrophysiological tests showed significant photopic b-wave amplitude decreases, particularly in type 3E (42.2% of normal). The electro-oculogram light peak/dark trough ratio was abnormal in 60 of 132 eyes (45.5%), especially in type 3E (25 of 34 eyes, 73.5%). Visual prognosis and overall visual function varied depending upon disease type, location of retinal lesions, and age of onset. In Type 3E, overall retinal function was poorest and accompanied by the most severe decline of central vision and function in the surrounding macula. PMID- 8224930 TI - Cytokines in the conjunctiva of acute and chronic mucous membrane pemphigoid: an immunohistochemical analysis. AB - The aim of this study was to evaluate the potential role of certain soluble factors in conjunctival scar tissue formation of pemphigoid patients. Epibulbar conjunctival biopsy specimens were taken from patients with acute ulcerative (n = 4), subacute (n = 8) and chronic (n = 8) mucous membrane pemphigoid and from twelve age-matched healthy individuals. The tissues were embedded in glycol methacrylate and analysed by immunohistochemical methods. Interleukin-2 (IL-2), interferon-gamma, transforming growth factor-beta (TGF-beta), platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), tumour necrosis factor-alpha and proliferating cells (as identified with the antibody Ki-67) were found in both pemphigoid patients and normal controls. Interleukin-4 was not found with this method in either normal or diseased conjunctiva. Significant differences between normal and diseased conjunctiva were found for TGF-beta and for proliferating cells, which were both increased in the acute disease group. More intense staining was found in the subacute disease group for IL-2, bFGF and PDGF. Our findings showed that a variety of cytokines were present in normal and diseased bulbar conjunctiva. Acute conjunctival disease in mucous membrane pemphigoid may indicate active scar tissue formation, implied by an increase in TGF-beta and the presence of proliferating fibroblasts. PMID- 8224931 TI - Nd:YAG laser trabeculopuncture (YLT) for glaucoma with traumatic angle recession. AB - Traumatic angle recession caused by blunt trauma often induces uncontrollable glaucoma despite the maximum medical therapy tolerated, such as argon laser trabeculoplasty (ALT), with no or little benefit. Therefore, instead of ALT, we tried Nd:YAG laser trabeculopuncture (YLT) on 11 patients with this type of glaucoma. The intraocular pressure of these patients was followed up for 15 +/- 7 months (average +/- SD). In 6 of 7 eyes treated initially with YLT, the IOP was significantly reduced, so medication was discontinued. Four other cases with uncontrollable IOP after failed ALT were treated with YLT. The IOP of 3 cases was successfully controlled by medication after YLT. These YLT results were then compared with those of ALT in 11 glaucoma patients with traumatic angle recession. Seven of 11 cases treated initially with ALT failed in less than 3 months, and surgical intervention or additional laser treatments were required. The probability of success from the time-series analysis at 12 months after each laser application was 0.909 in YLT, 0.273 in ALT. YLT offers significant advantages over ALT for the treatment of glaucoma with traumatic angle recession after blunt trauma and thus merits further study. PMID- 8224932 TI - Contrast sensitivity in patients with keratoconjunctivitis sicca before and after artificial tear application. AB - Poor stability of the precorneal tear film causes dry eye symptoms and has detrimental effects on the optical function of the eye. In 50 patients with dry eyes the local instillation of artificial tears significantly improved contrast sensitivity (P < 0.001), using the Vision Contrast Test System 6500. In the untreated eyes no significant changes were found to occur (P = 0.56). PMID- 8224934 TI - Immunocytochemistry of types I-IV collagen in human anterior subcapsular cataracts. AB - Human anterior subcapsular cataracts were examined by immunocytochemistry to investigate which types of collagen constitutes the subcapsular fibrous matrix. Types I, III, and IV collagen were shown to be immunologically positive by both light and electron microscopy. In light microscopy, types I and III collagen were stained diffusely whereas type IV collagen was stained in a streaky pattern in the opacities. The lens capsules were stained with antibody to type IV collagen, although the staining patterns were not homogeneous; the inner layers of the capsules were labeled more strongly than the outer layers. Neither type I nor III collagen was detected in the capsules. In electron microscopy, collagen fibrils were labeled with the antibodies to both type I and type III collagen. The lens capsules and multilamellae of the basement membranes in the opacities were labeled with the antibody to type IV collagen. PMID- 8224933 TI - Heritability of myopic refractive errors in identical and fraternal twins. AB - The existence of a visual feedback control of eye growth in humans is controversial, as the contributions of genetic and environmental factors are still unknown. To evaluate the heritability of refractive defects, we measured ocular refraction in 19 monozygote and 20 dizygote twin pairs (mean age 5 years). Monozygosity was ascertained by a common chorion, similarity of somatic traits, and identical dermatogliphes and was confirmed in myopes by blood marker diagnosis. Ocular refractive defects and axial length were evaluated by cycloplegic autorefractometry and biometry. By comparing identical and fraternal twins heritability of refractive defects was estimated to be 0.08-0.14; this low value indicates that the observed variability in refractive errors is nongenetic in origin. Three monozygote pairs were anisomyopic; differences between eyes in identical twins were related to the increased axial length of myopic eyes. In one eye, myopia was attributed to visual deprivation induced by a congenital cataract, while in five eyes it was correlated directly to the degree of astigmatic defects. The discordant axial length observed in monozygote twins is nongenetic. In agreement with previous findings reported in the literature, it is proposed that visual impoverishment of retinal images may play an early regulatory role in postnatal eye growth. PMID- 8224935 TI - Ocular toxicity of daunomycin: effects of subdivided doses on the rabbit retina after vitreous gas compression. AB - Daunomycin is a potent antiproliferative agent which has been shown to prevent experimental proliferative vitreoretinopathy. However, toxic effects on the rabbit retina have been reported even for the lowest effective doses. In a previous report we demonstrated that subdivided doses rather than single doses of daunomycin improves the efficacy in prevention of experimental proliferative vitreoretinopathy. To evaluate whether dose subdivision would also have an alleviating effect on drug toxicity, we administered 15 nmol daunomycin in doses of 10 nmol and 5 nmol 4 h apart into the vitreous cavity of rabbit eyes which had previously undergone vitreous gas compression. All contralateral eyes received sham treatment. Simultaneous electroretinographic recordings from both eyes on days 0, 3, 7, and 14 demonstrated a significant b-wave decline in drug-exposed eyes. Morphological studies on these eyes revealed no retinal damage. Our findings suggest that dose subdivision does not eliminate the retinal toxicity of daunomycin. PMID- 8224936 TI - Enhanced detection of normal retinal nerve-fiber striations using a charge coupled device and digital filtering. AB - We used a 1024 x 1024 pixel, 15-microns, 16-bit-encoding, multi-pin-phase charge coupled device (CCD) to obtain images of the normal human retinal nerve fiber layer. This device, which operates at room temperature, offers significantly better signal-to-noise ratio, linearity, and dynamic range than do photographic film, video imaging techniques, or commercially available CCDs. We demonstrate the use of a nonlinear digital filter, together with filter windows, that enhances fine detail of NFL striations, while suppressing noise, in limited areas of the CCD images. High-sensitivity imaging of this type, together with appropriate digital processing, may prove useful in diagnosing and following nerve-fiber-layer damage due to glaucoma. PMID- 8224937 TI - Spectral sensitivity of the blood-retinal barrier at the pigment epithelium for blue light in the 400-500 nm range. AB - To specify the spectral sensitivity of the retinal pigment epithelium (RPE) for blue light damage, pigmented rabbits were exposed to light of 408, 418, 439, 455, 485, and 500 nm (half-peak bandwidth approximately 12 nm). The range of radiant exposure was 15-275 J cm-2 (1.7-19 mW cm-2 for 0.5-5 h). Vitreous fluorophotometry was used to functionally evaluate the blood-retinal barrier at the RPE in vivo, and electron microscopy to visualize RPE ultrastructure in vitro. A significant increase in permeability of the blood-retinal barrier was seen only after exposure to light of 418 nm. Radiant exposure at threshold for permeability increase was 18 J cm-2. Electron microscopy of the RPE demonstrated dispersion and clumping of melanin granules. The results suggest that the RPE is most sensitive to light in the range 412-425 nm, possibly due to damage-mediating chromophores such as cytochrome c oxidase and lipofuscin. PMID- 8224938 TI - Ophthalmological findings in three patients with cholesterol acyltransferase deficiency syndrome before and after kidney transplantation. PMID- 8224939 TI - Color vision defect in Purtscher's disease. PMID- 8224940 TI - Glaucoma as a cause of poor vision in severe retinopathy of prematurity. AB - Despite surgical reattachment of the retina in infants with severe retinopathy of prematurity (ROP), visual acuity may decline. We performed gonioscopy and applanation tonometry on 26 eyes of 17 infants with severe ROP who were randomly selected and followed prospectively for 2 years. Of these 26 eyes, one eye of one patient had only peripheral retinocryopexy for threshold stage III disease and had an attached retina before enrollment into the study. Sixteen infants subsequently underwent vitrectomy, scleral buckle, or both. Retinal reattachment of the posterior pole was present in at least one eye each of 10 of the 17 infants. Glaucoma was diagnosed in 5 of the 10 patients; all 5 showed a decrease in visual acuity of greater than 1 octave. Of the 5 infants without glaucoma, 3 showed visual improvement, and 2 remained the same. Although we studied a small number of cases, our results suggest that glaucoma may be a cause of visual decline in infants with severe ROP. A decline in visual acuity in infants with severe ROP and surgical retinal reattachment may warn us of glaucoma, as this diagnosis is difficult to make in these small infants. PMID- 8224941 TI - Plasma glucose levels, postoperative complications, and progression of retinopathy in diabetic patients undergoing intraocular lens implantation. AB - Of 220 patients (250 eyes) who underwent extracapsular cataract extraction with implantation of a posterior chamber intraocular lens, 36 (45 eyes) with non insulin-dependent diabetes mellitus had no retinopathy, simple, or preproliferative retinopathy. They were divided into group A patients who underwent diet therapy or were treated with oral agents and group B patients who were given insulin therapy. The 184 patients (205 eyes) without diabetes mellitus served as control (group C). Transient elevation of intraocular pressure and cystoid macular edema were observed more frequently in diabetic patients (groups A and B). Progressive retinopathy occurred more frequently in group B than in group A patients, and it correlated with increased levels of plasma glucose. Unilateral cataract surgery was performed on 27 diabetic patients. Nine (33%) operated eyes showed progressive retinopathy, while one (4%) fellow eye worsened. It is possible that both increased levels of plasma glucose and surgical procedures may participate in the progression of retinopathy. PMID- 8224942 TI - Biomicroscopy versus fluorescein angiography of the iris in the detection of diabetic iridopathy. AB - Iris fluorescein angiography is not commonly employed in clinical practice, but it is the most sensitive technique for the evaluation of iris vessel abnormalities. We used iris fluorescein angiography as the gold standard against which to test the ability of iris biomicroscopy to demonstrate diabetic iridopathy (DI). One hundred and fourteen eyes of 63 diabetic patients affected by preproliferative or proliferative diabetic retinopathy (DR) (the DR groups at high risk of developing DI) were considered. The DI fluorangiographic classification used was: (1) absence of DI; (2) nonproliferative DI; (3) proliferative DI. The sensitivity of biomicroscopy in detecting DI turned out to be 57%, while the specificity was 94%. The positive predictive value was 93% and the negative predictive value 50%. Our study proved that biomicroscopy can accurately judge when DI is absent. When it is present, however, there is a high probability that biomicroscopy will be less precise in the detection of iris lesions. PMID- 8224943 TI - Functional vision with hydrogel versus PMMA lens implants. AB - As a particular test of visual function, contrast sensitivity reveals optical properties of Iogel pHema lenses in comparison to poly(methyl methacrylate) implants (PMMA). Sixteen patients with a hydrogel posterior chamber lens in one eye and a PMMA posterior chamber lens in the other were examined by means of contrast sensitivity measurements. Six stationary, vertical, sinusoidal modulated gratings with spatial frequencies of 0.5, 1, 3, 6, 11.4 and 22.8 cycles/degree were presented. Results of the two different implants were analyzed by a paired t test. There was no significant statistical difference between the two types of lenses for any of the gratings presented. Despite different features like material, refractive index, design and UV-absorbing additive, there seems to be no difference between lenses made of hydrogel and those made of PMMA material with regard to functional vision as evaluated by contrast sensitivity testing. PMID- 8224944 TI - Diagnostic problems in chronic vitreous inflammation. AB - We reviewed 188 patients with chronic vitreous inflammation initially diagnosed as intermediate uveitis, who were seen at the Ghent Eye Clinic during the period 1978 to 1988. We were able to establish a more specific diagnosis in 55 patients. The different approaches that contributed to the diagnosis were analyzed, (comprehensive history-taking and clinical examination, laboratory tests and special investigations, vitrectomy, and following the evolution of the disease). No etiology was found in 133 patients (70.7%), subsequently diagnosed as idiopathic intermediate uveitis. The major characteristics of the disease were analyzed. PMID- 8224945 TI - Reproducibility of topometric data acquisition in normal and glaucomatous optic nerve heads with the laser tomographic scanner. AB - The reproducibility of optic disc cup measurements was analyzed in 24 eyes of 24 patients [8 normals, 8 glaucoma patients, 8 glaucoma suspects] using the Laser Tomographic Scanner. The mean coefficient of variation in triple measurements was 5.0% for the cup area, 5.4% for the rim area, 7.0% for the cup volume, 4.0% for the mean cup depth, and 4.3% for the maximum cup depth. Mean reliability between two of the three measurements performed in each eye was better than 0.988 for the cup area, 0.995 for the cup volume and 0.996 for the mean cup depth readings. These results suggest that laser scanning tomography allows highly reproducible measurements in living eyes and adds an important tool to the ophthalmologist's armamentarium for the diagnosis and follow-up of glaucoma patients. PMID- 8224946 TI - Evaluation of FASTPAC: a new strategy for threshold estimation with the Humphrey Field Analyser. AB - A new strategy for threshold estimation, FASTPAC, has been introduced for the Humphrey Field Analyser with the aim of improving the efficiency of visual field examination without loss of accuracy. FASTPAC was compared to the standard 4-2 double staircase strategy in 98 normal volunteers (age 23-83 years). One eye of each subject was examined with both strategies on two visits using Program 30-2. The order of strategy was randomised. FASTPAC examination time was 43% faster than the standard algorithm, but the mean sensitivities were similar. Short-term fluctuation was 24% higher with FASTPAC (P < 0.001). Most parameters were adversely affected by increasing age. FASTPAC offers a valid alternative to the standard strategy in the examination of suspected normal fields and should always be considered as an alternative to the threshold-related screening strategies. The performance of the algorithm in the identification of field loss and in areas of reduced sensitivity awaits further study. PMID- 8224947 TI - Basal laminar deposit in the aging peripheral human retina. AB - A basal laminar deposit (BLD) in the human macula has been described as an early sign of age-related macular degeneration. In some eyes with a BLD in the macula, light microscopic sections of the peripheral retina revealed almost similar deposits between the retinal pigment epithelium and Bruch's membrane. Because the exact pathogenesis of age-related macular degeneration and the origin of the BLD are unknown, we studied the ultrastructure of these peripheral sub-RPE deposits. Parts of the equatorial and peripheral regions of the retina of ten human eyes, with BLD-like deposits between the retinal pigment epithelium and Bruch's membrane, were examined by electron microscopy. In eight of these ten eyes the ultrastructure of these deposits was amorphous and finely granular. Five of the eight deposits also contained small amounts of long-spacing collagen. Ultrastructurally, the deposits were similar to an early type BLD in the macula. In the remaining two eyes, the deposits appeared to consist of flat, elongated drusen. Our findings indicate that a BLD can develop not only in the macula but also in the peripheral region of the retina. PMID- 8224948 TI - A new model of transient complete obstruction of retinal vessels induced by endothelin-1 injection into the posterior vitreous body in rabbits. AB - Endothelin-1 (ET-1) is a potent vasoconstrictor peptide produced by vascular endothelial cells. In order to investigate the effects of ET-1 on retinal vessels, ET-1 (1-1000 pmol) was injected into the posterior vitreous body in rabbits. A high dose of ET-1 induced transient complete obstruction of the retinal vessels. In this experimental model of transient complete obstruction of the retinal vessels, the effects of ET-1 on retinal function were further analyzed by means of electroretinograms. The scotopic a-wave was not affected, but the amplitude of the scotopic b-wave was significantly elevated. The amplitude of oscillatory potentials was significantly reduced. These phenomena suggested that retinal ischemia without choroidal ischemia was brought about due to severe vasoconstriction of the retinal arteries. These findings indicate that intravitreal injection of ET-1 causes a transient cessation of blood supply from retinal vessels and that oscillatory potentials in electroretinograms appear to be sensitive for detecting changes of retinal circulation. This new model of transient complete obstruction of retinal vessels might be useful for studying the pathophysiology of severe retinal ischemia. PMID- 8224949 TI - Indoleamine 2,3-dioxygenase activity in the aqueous humor, iris/ciliary body, and retina of the bovine eye. AB - The enzyme indoleamine 2,3-dioxygenase (IDO), which uses free oxygen radicals to cleave the pyrrole ring of indoleamines and give kynurenamines, has previously been found in most tissues, but not in the eye. In this study, IDO activity was measured in post-mortem bovine eyes using Yamazaki's method with L-tryptophan as substrate. Because of the physiological importance of IDO in the protection against free oxygen radical damage, a search was conducted to find this enzyme in the eye. Products of tryptophan degradation by IDO, the kynurenine and 3 hydroxyanthranilic acid were detected and measured in the aqueous humor, iris/ciliary body, and the retina by high-performance liquid chromatography (HPLC) coupled with electrochemical detection. IDO activity was 3.2, 9.0 and 10 nmol/mg protein per h for the aqueous humor, iris/ciliary body, and retina, respectively. These findings suggest that, because of its scavenger properties, IDO is involved in the protection of the eye where, because of its transparency, free radicals are formed not only in the normal oxidation process, but also photochemically. PMID- 8224950 TI - The diameter of the ciliary sulcus: a morphometric study. AB - Based on a precise definition of boundaries of the ciliary sulcus, a new anatomical method was used to measure the diameter of the ciliary sulcus in postmortem eyes. In each of 41 globes, 24 different ciliary sulcus diameters were measured and were compared with the axial length, measured by ultrasonography. The mean ciliary sulcus diameter was 11.25 mm (SD = 0.38 mm). By linear regression, the axial length values were significantly correlated with ciliary sulcus diameters. In addition, our data suggest that existence of two groups of eyes differentiated exclusively by the diameter of the ciliary sulcus, independent of the axial length. We propose that the displacement of the pars plicata corporis ciliaris from the peripheral iris to the iris root during the embryonic period may be incomplete in eyes of shorter axial length. The findings of the present study may play a role in cataract surgery in which an intraocular lens is inserted into the ciliary sulcus. PMID- 8224951 TI - Diode versus argon-green laser panretinal photocoagulation in proliferative diabetic retinopathy: a randomized study in 44 eyes with a long follow-up time. AB - A clinical randomized study using argon-green (532 nm) and diode (810 nm) lasers was carried out on 44 eyes (34 patients) affected by proliferative diabetic retinopathy (PDR), with the aim of evaluating the long-term effects of diode panretinal photocoagulation. Of the total 44 eyes, 22 were randomized to argon laser treatment (ALT) and 22 to diode laser treatment (DLT). The mean follow-up time was 25 +/- 5 months in the ALT group versus 24 +/- 4 months in the DLT group. In the ALT group 20 eyes (91%) showed regression of PDR whereas 2 eyes (9%) deteriorated. In the DLT group regression of neovascularization was observed in 22 eyes (100%). These results show that the long-term efficacy of diode laser PRP in the treatment of PDR is similar to that of argon-green laser PRP. PMID- 8224952 TI - Sophisticated vitreoretinal surgery in patients with a healthy fellow eye. An 11 year retrospective study. AB - Forty-four patients who had undergone successful vitreoretinal surgery were included in a retrospective study intended to evaluate the real benefit of surgery in each case. All patients presented with a retinal detachment complicated by proliferative vitreoretinopathy in one eye, the fellow eye being healthy. The number of operations undergone by each patient and the total operating time per patient were carefully recorded. Objective parameters such as visual acuity, visual field and stereoscopic function were monitored. Visual comfort and subjective complaints were graded by means of a questionnaire given to the patients. Any severe event occurring in the fellow eye during the follow up period was also registered. Sophisticated vitreoretinal surgery in patients with a good fellow eye is useful to the patient if binocular function can be restored at least partially. Correction of unilateral aphakia by a contact lens should not be considered an acceptable solution. PMID- 8224953 TI - Combined iodine-125 plaque irradiation and indirect ophthalmoscope laser therapy of choroidal malignant melanomas: comparison with iodine-125 and cobalt-60 plaque radiotherapy alone. AB - The authors studied the short-term impact of combined episcleral iodine-125 plaque radiotherapy and argon laser treatment in a series of 24 patients with choroidal malignant melanoma. All patients underwent plaque therapy prior to their initial laser session. All laser treatments were performed with an indirect ophthalmoscope argon green laser, using low-power, long-duration exposures. The endpoint of laser therapy was a well-defined atrophic circumbasal chorioretinal laser scar and complete or nearly complete nonfluorescence of the lesion on fluorescein angiography. In a case-by-case matched comparison study, the authors evaluated the relative local regression of tumors treated by combined plaque laser therapy, iodine-125 plaque therapy alone, and cobalt-60 plaque therapy alone. The tumors treated with supplemental laser regressed substantially faster and more completely than did those treated by either type of plaque therapy alone. However, the short-term visual loss was greater in eyes treated by the combined therapy. PMID- 8224954 TI - An improved technique for the diagnosis of viral retinitis from samples of aqueous humor and vitreous. AB - We applied the technique of DNA amplification with the polymerase chain reaction to nine aqueous humor and five vitreous samples from HIV-1-infected patients with clinically diagnosed cytomegalovirus retinitis. For the amplification, recently published primers specific for herpes simplex virus (HSV), varicella zoster virus (VZV) and cytomegalovirus (CMV-1) were used. Additionally, a newly developed primer pair specific for the main immediately early gene of CMV (CMV-2) was selected and compared with the published one. All primers were tested on noninfected and HSV-, VZV- and CMV-infected human fibroblast cell culture supernatant, thereby excluding cross-reactivity of the chosen primers. In none of 13 aqueous humor and six vitreous samples of healthy controls was any viral DNA amplified. Using the CMV-1 primers, we detected CMV DNA in five of nine aqueous humor and three of five vitreous samples amplifying a DNA fragment 435 base pairs in length. With the CMV-2 primers, we detected a CMV DNA fragment with a length of 110 base pairs in eight of nine aqueous humor and in four of five vitreous samples. Additionally, CMV DNA was found in three of nine urine and two of nine saliva specimens. Both CMV and HSV DNA were amplified in one aqueous sample. Varicella DNA was not detected in any of the specimens. Thus, the polymerase chain reaction is more sensitive than other comparable diagnostic tests and may provide an alternative to conventional virus isolation and in situ hybridization techniques for the laboratory diagnosis of viral ocular disease. PMID- 8224955 TI - Performance of two films for densitometry of retinal photographs. AB - The relative performance was determined of two different photographic films (Kodak Panatomic-X and Kodak Technical Pan) for densitometry readings from retinal photographs of patients who had or were suspected of having glaucoma. The raw data from the two films were significantly different. The high contrast film was particularly sensitive to external variables; however, when normalised in terms of the standardised deviation, the data from the two films were comparable (P < 0.005). We also measured the pattern of nerve fibre layer loss using digital image analysis of red-free photographs of normal, ocular-hypertensive, glaucoma suspect and glaucomatous eyes. Several mathematical techniques were used to characterise the data from each eye and then to compare these data to the mean photographic density of the normal eyes. Results showed that it was possible to separate normal eyes from glaucomatous eyes. The highest sensitivity achieved was 100% for right and 88% for left eyes, and the highest specificity was 100% for both eyes. PMID- 8224956 TI - The role of Pseudomonas aeruginosa elastase in corneal ring abscess formation in pseudomonal keratitis. AB - In order to identify the causative factors of ring abscess, which is the characteristic feature of pseudomonal keratitis, pseudomonal endotoxin, exotoxin A, and elastase were each separately injected into guinea pig cornea. There was no formation of ring abscess. Injection of living Pseudomonas aeruginosa strains IFO3455 and Takamatsu which produce all three molecules, clearly induced ring abscess. In contrast, when heat-killed bacteria strain IFO3455 or living bacteria of the non-elastase-producing strain PA103 were injected, ring abscess was not induced. Furthermore, when living bacteria strain IFO3455 were injected with anti elastase antibody or a protease inhibitor, ovomacroglobulin, ring abscess formation was significantly inhibited. Histological examination demonstrated that the ring abscess was a dense accumulation and aggregation of polymorphonuclear leukocytes (PMN) with debris of cells and lamellae in the deep stroma at the corneal margins, suggesting prevention of PMN migration to the central lesion. The presence of anti-elastase antibody or a specific elastase inhibitor facilitated PMN migration towards living bacteria strain IFO3455 in an in vitro model. These results indicate that pseudomonal elastase is a necessary but not sufficient factor in the formation of ring abscess in pseudomonal keratitis. PMID- 8224957 TI - Latent endothelial cell damage after experimental corneal cryopreservation. AB - Ninety porcine corneas were evaluated by vital staining with alizarin red S and trypan blue in a three-step experiment. Central cell densities were counted (a) on freshly dissected corneas (n = 30), (b) on cryopreserved corneas directly after thawing (n = 30), and (c) after a postthawing organ culture interval of 24 h (n = 30). Two freezing methods were used: (a) minimum essential medium- containing 20% fetal calf serum and (b) the same but containing additionally 2% chondroitin sulfate. Directly after thawing neither method showed significant cell loss (3.9% and 3%) compared to fresh tissue. After postthawing organ culture, however, tissue that had been frozen without chondroitin sulfate displayed a cell loss of 73.5% compared to corneas of the same freezing protocol directly after thawing. Corneas in chondroitin sulfate containing medium showed a cell loss of only 33.2%. We conclude that reliable morphologic evaluation should not be obtained from cryopreserved corneas examined directly after thawing. PMID- 8224958 TI - Histochemical survey of the anterior segment of the normal human foetal and adult eye. AB - The distributions of the lysosomal enzymes [acid phosphatase (AP), N-acetyl-beta D-glucosaminidase (NAG), beta-glucuronidase (beta-Gluc), beta-galactosidase (beta Gal), dipeptidylpeptidase II (DPP II)] and of the membrane-bound proteases [aminopeptidase M (APM), aminopeptidase A (APA), gamma-glutamyltransferase (GGT), dipeptidylpeptidase IV (DPP IV)] were investigated in the normal human adult and foetal anterior segment by histochemical methods. The distribution of these hydrolases varied between ocular tissues. The most active enzymes in the adult corneal epithelium and endothelium were AP, beta-Gluc, NAG, beta-Gal and GGT; in the keratocytes, APM, APA, beta-Gluc and GGT predominated. The adult trabecular meshwork cells were stained by AP, beta-Gluc, NAG, APM, GGT, DPP II and DPP IV. The enzymes AP, beta-Gluc, APM and APA, however, displayed greater activity in the endothelium of Schlemm's canal. The adult ciliary epithelium stained strongly for all lysosomal hydrolases; GGT was the most active protease here. Differences in enzyme activity were noted in some tissues when foetal and adult anterior segments were compared. There appeared to be a decrease in the activity of some enzymes with age and post-mortem delay greater than 24 h. The function(s) of each enzyme and their possible roles in the respective tissues are discussed. PMID- 8224959 TI - Vasoactive intestinal peptide/peptide histidine isoleucine mRNA in the eye and suprachiasmatic nucleus of normal and monocularly enucleated rats. AB - The localization of the messenger RNA (mRNA) encoding vasoactive intestinal peptide/peptide histidine isoleucine (VIP/PHI) in the rat eye was studied by in situ hybridization histochemistry using a synthetic 35S-labeled oligodeoxyribonucleotide. Among the layers of the retina, specific labeling was found in the soma of some cells in the innermost lamina of the inner nuclear layer. Occasionally, labeled cells were also present in the ganglion cell layer. No specific labeling was detected in any ocular structures other than the retina, including the cornea, iris, ciliary body, choroid, choriocapillaris, optic nerve and sclera. We also examined VIP/PHI mRNA expression in the suprachiasmatic nucleus (SCN) of the hypothalamus, which is the pacemaker of circadian rhythms and synchronizes them with the environmental light-dark cycle. High labeling was found in the ventrolateral part of the nucleus, which is the site of most projections from the retina. Monocular enucleation of rats at birth resulted in an increase of VIP/PHI mRNA expression in SCN ipsilateral to the removed eye after postnatal day 60. The discrete pattern of hybridization for VIP/PHI mRNA indicates that these two peptides are synthetized in rat retina and SCN and suggests that they may play a key role in the photic entrainment to the SCN regulating various circadian rhythms. PMID- 8224960 TI - Metastasis of malignant melanoma of the choroid and ciliary body to the contralateral choroid. PMID- 8224961 TI - No changes in rabbit corneal esterase activities with dipivefrine hydrochloride instillation for 4 weeks. PMID- 8224962 TI - Jacques Joseph. PMID- 8224963 TI - Sigmund Freud's illness--the ultimate team approach to head and neck cancer? PMID- 8224964 TI - Historical trends in surgery for the aging face. PMID- 8224965 TI - The history of craniofacial surgery. PMID- 8224966 TI - The history of oculoplastic surgery. PMID- 8224967 TI - Facial surgery during World War I. PMID- 8224968 TI - The early history of rhinoplasty. PMID- 8224969 TI - A brief sortie into the history of cranio-oculofacial prosthetics. PMID- 8224970 TI - [Problems in providing medical information for ecologic research]. PMID- 8224971 TI - [Evaluating the connection between health parameters of the population and environmental quality]. PMID- 8224972 TI - [Hygienic mapping in the system of environmental monitoring]. PMID- 8224974 TI - [Methods of forming combined indicators in ecologic and hygienic investigations]. PMID- 8224973 TI - [Hygienic approaches to radio ecologic mapping of industrially polluted regions]. PMID- 8224975 TI - [Methodologic approaches to evaluation of water factor effect on population health using the example of regions with developed cellulose industry]. PMID- 8224976 TI - [Combined indicator in the assessment of respiratory organs in exposure to dust]. PMID- 8224977 TI - [Integrated evaluation of the risk factors for recurring bronchitis in children]. PMID- 8224978 TI - [Scientific basis for mapping of electromagnetic environmental pollution]. PMID- 8224979 TI - [Ecologic and hygienic aspects of regulation of the maximum permissible level of water contamination]. AB - The research has shown that in calculation of the permissible load of a water body all compounds should be taken into account whatever their harmful limit indicator. Total loading coefficient at a level below I in all cases does not guarantee a negative influence on auto-purification process. PMID- 8224980 TI - [Microbiological evaluation of water pollution in the Volga delta]. AB - Bacterial contamination of water and fish in the Volga delta region was studied. Water near large enterprises contained more bacteria than at 75 km a distance of downstream. All organs and tissues of sturgeon contained opportunistic bacteria. Bacterial contamination of water is responsible incidences for acute intestinal diseases of the population. PMID- 8224981 TI - [Sanitary aspects of the dynamics of water quality of the Kronshtadt water supply system]. PMID- 8224982 TI - [Toxicologic evaluation of purified municipal and industrial waste water]. AB - Analysis of waters sewage in Kiev and of waste water of a textile and se wing enterprise in Chernigov has shown that treatment by biological method and with activated carbon was fairely efficient in toxicity reduction. PMID- 8224983 TI - [Hygienic principles of evaluating the air pollution level]. PMID- 8224984 TI - [Effect of industrial noise and ototoxic antibiotics on ear function in man]. AB - Fifty workers of the 166 ones examined were occupationally exposed to noise; 26 of them were treated with ototoxic antibiotics. Workers of treated with antibiotics most often (69.2%) developed deterioration of acoustic function. PMID- 8224985 TI - [Staphylococcus carriers in workers of the Astrakhan gas condensing plant]. AB - Frequency of staphylococcus carriership in upper respiratory route mucosa of workers engaged in gas industry depends on intensity and duration of occupational contact with chemical pollutants of the working environment. PMID- 8224986 TI - [Rationale for the maximum permissible concentration of porophor P-18-S in the air of work place]. AB - Porophor P-18-S LD50 is 712.5 mg/kg. This substance is not characterized by cumulative or irritating properties. It has an expressed neurotropic and allergenic effects, can induce anemia and leukocytosis in animals. Work with this substance is allowed on condition of eye and respiratory route protection. Its MAC for working environment is 3.08 mg/m3. PMID- 8224987 TI - [Working conditions and morbidity in workers of hydrolysis yeast industry in the Uzbekistan Republic]. AB - The main occupational hazards in hydrolysis yeast industry of Uzbekistan are protein dust, fungi producers, adverse physical factors. Morbidity of factory workers is higher than in control group (office employees). Infectious diseases of upper respiratory tract, influenza, bronchitis, broncho-tracheitis are prevalent in this factory. PMID- 8224988 TI - [Measurement of trace quantities of plant protective biological agents in vegetables and their toxicologic assessment]. PMID- 8224989 TI - [Effect of tutorial computer lessons on the functional status of 7-13 years old children]. AB - First- and fifth-year pupils experience the greatest difficulties in adaptation to computer activity. The most tiresome for pupils are computer games, computer programming ranks second. Mixed type lessons are the most favourable for functional state of pupils. PMID- 8224990 TI - [Study of the anticarcinogenic characteristics of the trace element, selenium, sanitary-hygienic experiment]. AB - Enrichment of rat daily diets by 1-2 (up to 5) mg selenium increased nonspecific resistance of animals to ionizing radiation. Incidence of mammary, pituitary, and thyroid tumors as well as of leukemia were lower as compared with control animal group given no selenium. PMID- 8224991 TI - [Radionuclide inhalation during agricultural work in the regions polluted by the Chernobyl accident]. AB - With pollution of agricultural fields by plutonium at a level of more than 3.7 kBg/m2 and cesium more than 7.4 MBq/m2 radionuclide levels in tractor cabin can exceed the permissible level. However field work being seasonal, annual level of inhaled radionuclides does not exceed the permissible limit. Actually the equivalent dose for the lungs does not exceed just few mSv under conditions of agricultural production. PMID- 8224992 TI - [Some methodologic problems of prenosologic hygienic diagnosis]. PMID- 8224993 TI - [Hygienic evaluation of ozone utilization for the sanitation of tobacco polluted indoor air]. PMID- 8224994 TI - [Age factors in toxicologic and hygienic studies (review)]. PMID- 8224995 TI - [Problems in training of specialists at institutes of medical prevention due to the reorganization of sanitary and epidemiologic service]. PMID- 8224996 TI - [The work of Grigory Vitalevich Khlopin in the Ukraine (90 years anniversary of the foundation of hygiene department in Odessa)]. PMID- 8224997 TI - [Changes of functional parameters in experimental animals in relation to length of swimming]. PMID- 8224998 TI - [Gas chromatographic determination of some aromatic compounds in water by analyzing steam phase balance]. PMID- 8224999 TI - [Comment on the article "Tasks of physician-hygienist training today"]. PMID- 8225000 TI - [Prognosis of accumulative characteristics of chemical substances based on using acute experimental data]. PMID- 8225001 TI - [Hygienic principles of industrial waste assessment for cartography]. PMID- 8225002 TI - The aging mind: potential and limits. AB - Research on the aging mind has moved from a simple growth vs decline view to a conception of a joint consideration of potential and limits. This development is illustrated by research on two categories of cognitive functioning: the cognitive mechanics and the cognitive pragmatics. The cognitive mechanics (comparable to fluid intelligence) are primarily determined by the evolution-based neurophysiological architecture of the mind, whereas the cognitive pragmatics (comparable to crystallized intelligence) primarily reflect the impact of culture. Testing-the-limits research on basic memory serves as a prototypical instantiation of the aging of cognitive mechanics; research on wisdom and the cognitive management of the self are prototypical examples of the aging of cognitive pragmatics. In the cognitive mechanics, as one would expect from a phenomenon which is largely genetically and biologically controlled, there is definite aging loss. Conversely, in the cognitive pragmatics, which is primarily culture-based, there is evidence for stability and positive change in persons who reach old age without specific brain pathology, and who live in favorable life circumstances. A model of successful aging, selective optimization with compensation, is presented to illustrate how individuals and societies might effectively manage the age-related shift toward a less positive balance between gains and losses and the associated dynamics between culture-based growth and biology-based decline in level of functioning. PMID- 8225003 TI - Out-of-pocket health expenditure differentials between elderly and non-elderly households. AB - Regression analysis was used to compare out-of-pocket health expenditures between lower and higher income elderly and non-elderly households. Race and housing tenure were significant determinants for both age groups, but assets, education, and family size were significant only for the non-elderly. Analysis of out-of pocket health costs indicated greater equity among elderly persons, probably due to Medicare, but income elasticities revealed that health spending is a luxury for lower income elderly households. PMID- 8225004 TI - Gender differences in factors associated with change in physical functioning in old age: a 6-year longitudinal study. AB - We compared 356 older men and women and found that income, education, and marital status had significantly stronger associations with 6-year change in functioning for men; internal health locus of control was stronger for women. Large but not significant differences occurred for age, perceived health, and days going out per week (stronger for women) plus exercise and smoking (stronger for men). We found no difference for ethnicity, chronic conditions, and social contacts. Gender specific models incorporating factors amenable to change predicted significantly higher follow-up functioning for those with positive health behaviors, supporting the value of preventive interventions. PMID- 8225005 TI - A multivariate analysis of disability and health, and its change over time in the National Channeling Demonstration data. AB - The Channeling Demonstration examined the effects of case management interventions on a variety of outcomes. In the study, longitudinal data were collected from interviews of cases and controls. A multivariate procedure applied to this data identified groups with specific health profiles. Six profiles described health variation over individuals, and time, according to likelihood ratio statistics. Six sets of life tables were calculated, one for each health profile, to estimate the average duration of service use and the "follow-up" services used. A number of differences, and changes, in service use between the six groups were significant. PMID- 8225006 TI - Intrapersonal resources and the effectiveness of self-help groups for bereaved older adults. AB - The relative impact of three intrapersonal resources (self-esteem, competencies and life satisfaction) and the duration of a self-help group intervention on levels of depression and grief were examined over time among a sample of 295 recently bereaved older adults. In general, the resources examined had a greater direct influence on outcomes than the intervention. Also, those with lower competencies reported reduced depression and grief with greater meeting attendance, whereas the opposite was at first true for those with high competencies. Depression eventually decreased, however, among those with high competencies who continued to attend self-help group meetings for more than 8 weeks. Although bereavement self-help groups can aid those who may be deficient in selected skills, it is equally important to emphasize ways the bereaved can more effectively use those strengths that they may already possess in order to facilitate adaptation. PMID- 8225007 TI - To have or have not: parent status and the subjective well-being of older men and women. AB - Data from a multi-stage quota sample of adults aged 55 and over were used to examine the effect of parent status on three measures of subjective well-being- avowed happiness, depression, and satisfaction with life. Unlike other studies, we employed four categories of parent status: close parents, distant parents, those who chose not to have children, and those who are childless by circumstance. We found significant differences in subjective well-being between close parents and distant parents, and between close parents and the childless by circumstance but not the childless by choice. We also found important differences, by gender, in which factors predict subjective well-being. PMID- 8225008 TI - Changes in semantic memory in early stage Alzheimer's disease patients. AB - The types and number of exemplars of categories that are retrieved from semantic memory differentiate elderly normal controls and early stage Alzheimer's disease (AD) patients. Elderly normal controls generated more uncommon exemplars from closed semantic categories (fruits and vegetables) than did AD patients 2 1/2 years prior to the presumed onset of AD. AD patients, however, were just as productive as elderly normal controls in generating associations to open categories (letters). The findings suggest that one of the early cognitive symptoms of AD is changes in availability of uncommon exemplars of semantic networks. PMID- 8225009 TI - Identifying hearing loss and hearing handicap among chronic care elderly people. AB - This research assessed the appropriateness of the 1989 ASHA Draft Guidelines for the Identification of Hearing Impairment/Handicap in Adult/Elderly Persons for an institutionalized population. We tested 104 elderly chronic care residents using a conventional audiometer, an AudioScope3TM screener, and the Hearing Handicap Inventory for the Elderly. Results showed that this population is capable of performing these tests. Screening should be done at 2,000 Hz only. However, the prevalence of hearing loss is so high (69%) that we recommend conventional testing of all institutionalized elderly where feasible. PMID- 8225010 TI - Predicting the frequency of senior center attendance. AB - The frequency of senior center attendance was studied using the 1984 Supplement on Aging of the National Health Interview Survey. Unlike previous research, which focused on whether people ever attend a senior center, this study estimates a multinomial logistic regression model to distinguish between persons who rarely, sometimes, and frequently attend. There is evidence that the more frequent users are older, which may indicate difficulty in recruiting new active members. Greater frequency is associated with lower income and lower education but is unrelated to functional disability. The most frequent users are persons who are more socially involved. Finally, those who live in rural areas are more likely to be only occasional users, which may reflect a lack of accessibility or less frequent program availability in those areas. PMID- 8225011 TI - The continuum of care revisited: a rural perspective. AB - This study examines resource data for a variety of services in North Carolina's continuum of care for older adults. It qualifies the widespread contention that older adults in urban areas receive a greater number and more varied services than their rural counterparts. It also demonstrates that one of the most important differences between these communities is the greater variability of services among rural compared to urban counties. The comparative findings on urban and rural variations in the continuum of care for older adults have implications for further research and future policy development. PMID- 8225012 TI - A primary care nursing model in long-term care facilities: evaluation of impact on affect, behavior, and socialization. AB - A primary care model of delivery of nursing aide care was implemented and evaluated in one small, rural nursing home and one large, urban facility. Experimental unit residents received primary care consisting of permanent assignment of nursing aide, a team approach and enhanced communication. The urban experimental residents improved significantly in behavior and affect as measured across several sources, while the comparison group declined or remained the same. Significant improvements in behavior and social activities were observed among the experimental rural nursing home residents as contrasted with the comparison group after implementation of primary care nursing. It was concluded that primary care nursing as applied to nursing attendants in long-term care is beneficial to residents in terms of decreasing disturbed behavior and improved affect. PMID- 8225013 TI - Kinkeeping and friend keeping among older women: the effect of marriage. AB - Based on interviews with widows and wives, this article analyzes the effects of marriage and widowhood on older women's help to kin and friends. Although married women provide more help to kin than do widows, most of these differences can be explained in terms of the greater material resources marriage provides. In contrast, even controlling for other social characteristics, widows spend more time and give more practical help, in particular, to more friends than do wives. In two senses, then, marriage privatizes women's help to others--it provides them with both the resources and opportunity to help those related, while it reduces help--both its breadth and intensity--to those not related. PMID- 8225014 TI - Group homes for people with dementia: a Swedish example. AB - We describe the development and evaluation of group homes for dementia patients in Sweden. Group homes have been established on a social model of care to provide a better quality of life than in institutional settings, while making a small savings on cost. An enriched program of activities uses familiar, everyday household tasks, with staff trained to model appropriate behaviors. An evaluation of four homes indicated that residents could be well accommodated in the setting, even as their disabilities increased. Although there were some initial problems, staff satisfaction was high and turnover low. PMID- 8225016 TI - 46th Annual Scientific Meeting of the Gerontological Society of America. New Orleans, Louisiana, November 19-23, 1993. Abstracts. PMID- 8225015 TI - Developing a daily money management service model: navigating the uncharted waters of liability and viability. AB - This article discusses the development and implementation of a Daily Money Management (DMM) Service within a mature health and social service agency. Although there is increasing recognition of the need for DMM programs, major obstacles to their development include problems of securing adequate financial support and concerns about the liability risks inherent in providing financial services. This article describes an innovative DMM service model developed at Senior Health and Peer Counseling in Santa Monica, California, that has serviced more than 100 elders during its first two years of operation. The article explores the mission, key components and organization of the service model, including the development of a multidisciplinary Technical Advisory Group and a professionally supervised volunteer staff, the service planning and care coordination issues, the risk management protocols, and the need for multiple sources of financial support. PMID- 8225017 TI - A comparative evaluation of thiobarbituric acid methods for the determination of malondialdehyde in biological materials. AB - A comparative evaluation was made of the conventional spectrophotometric procedure and three published high performance liquid chromatographic (HPLC) procedures for the determination of malondialdehyde (MDA) as the thiobarbituric acid (TBA) derivative when applied to liver, fish meal, serum, and urine. Except for urine, spectrophotometric analysis overestimated MDA content. Purification of the TBA-MDA complex obtained from liver and fish meal on reverse phase cartridges was found to entail a loss of complex bound to residual peptides in the trichloracetic acid (TCA) extract. Mincing as opposed to homogenizing liver samples led to a doubling of values for MDA content. Hexanal was a major TBA reactant, in addition to MDA, in all the samples. Acid hydrolysis and heat were necessary for the release of MDA bound to the amino groups of proteins and other amino compounds. Methods for free MDA have limited application to biological materials except short term in vitro preparations such as peroxidizing microsomes, in which free MDA accumulates. On the basis of these and other observations, a modified HPLC procedure for the determination of MDA as the TBA MDA complex is proposed. PMID- 8225018 TI - 4-Hydroxynonenal, a metabolite of lipid peroxidation, activates phospholipase D in vascular endothelial cells. AB - We have examined the activation of phospholipase D (PLD) in bovine pulmonary artery endothelial cells (BPAEC) treated with 4-hydroxynonenal (4-HNE). Treatment of BPAEC labelled with [32P] orthophosphate (5 h for minimal phospholipid labelling) and [3H] myristic acid (24 h) with 4-HNE in the presence of 0.5% ethanol resulted in the formation of [3H] phosphatidylethanol (PEt) and [3H] phosphatidic acid (PA) with very little accumulation of [32P] PEt. The formation of [3H] PEt, as opposed to [32P] PEt, suggests that PEt synthesis was not through de novo pathway but rather through the PLD mechanism. 4-Hydroxynonenal-induced PLD activation was dose and time dependent, and was not associated with cytotoxicity as determined by [3H] deoxyglucose release. The formation of PEt was not affected by chelation of either extracellular Ca2+ with EGTA (5 mM, 30 min) or intracellular Ca2+ with BAPTA-AM (25 microM, 30 min). Treatment of BPAEC with either staurosporine (10 microM, 15 min), a protein kinase C (PKC) inhibitor, or down regulation of PKC by chronic 12-0-tetradecanoylphorbol-13-acetate (TPA) treatment (100 nM, 18 h) had no effect on 4-HNE-induced PLD activation. These results indicate that PLD activation by 4-HNE is independent of PKC activity. We also examined the specificity of nonylaldehyde derivatives and hydroxyalkenals on PLD activation. In addition to 4-HNE, 4-hydroxyoctenal and 4-hydroxyhexenal also stimulated [32P] PEt formation. Among the various nonylaldehydes examined, only trans-2-nonenal and trans-2-cis 6-nonadienal exhibited PLD activation, suggesting the requirement of a trans double bond at carbon 2 and a hydroxyl group at carbon 4. However, in contrast to 4-HNE-induced PLD activation of BPAEC monolayers, treatment of 105,000 x g membranes with 4-HNE had no effect on PLD catalyzed hydrolysis of [2-14C] oleoyl phosphatidylcholine. These data provide evidence that 4-HNE, a metabolite of membrane lipid peroxidation, may be involved in endothelial cell signal transduction, through the activation of phospholipase D and the generation of second messengers like phosphatidic acid and diacylglycerol. PMID- 8225019 TI - Consumption of carotenoids in photosensitized oxidation of human plasma and plasma low-density lipoprotein. AB - Human plasma and plasma low-density lipoprotein (LDL) were exposed to photoirradiation in the presence of methylene blue (water-soluble photosensitizer) or 12-(1-pyrene)dodecanoic acid (P-12, lipid-soluble photosensitizer). In methylene-blue-sensitized photooxidation of human plasma and LDL, endogenous carotenoids and tocopherols were consumed with the accumulation of cholesteryl ester hydroperoxides (CE-OOH). Xanthophylls (zeaxanthin and lutein) decreased faster than lycopene and carotenes in the case of human plasma. In P-12-sensitized photooxidation of human plasma and LDL, the decrease rate of xanthophylls was slower than that of lycopene and carotenes. A lower level of beta-carotene exerted the effective inhibition of lipid peroxidation and retarded the oxidative loss of alpha-tocopherol, when the phosphatidylcholine liposomes containing these two lipid-soluble antioxidants were subjected to methylene blue- or P-12-sensitized photooxidation. These results suggest that antioxidant activity of carotenoids in photosensitized oxidation (Type II) of human plasma LDL depends on the site of singlet oxygen (1O2) to be generated and that carotenoids can protect tocopherols from the oxidative loss by 1O2 in the plasma. PMID- 8225020 TI - Free radical intermediates in the oxidation of flavone-8-acetic acid: possible involvement in its antitumour activity. AB - The sulphate radical (SO4.-), a model one-electron oxidant, reacts with the antitumour drug flavone-8-acetic acid (FFA) with the rate constant 9.1 x 10(8) dm3mol-1s-1 to yield an uncharged radical that reacts with oxygen (k approximately 1 x 10(9) dm3mol-1s-1). The oxidation of FAA by SO4.- in a steady state system was found to release carbon dioxide with a yield of 96% relative to that of the SO.4-. The results are interpreted by fast (t1/2 < or = 1 microsecond) and efficient decarboxylation of the FAA radical cation, resulting in a carbon-centred radical. The reaction of the latter with oxygen is a possible source of radical-driven cytotoxic pathways, such as singlet oxygen formation via the Russell mechanism or H-abstraction from lipids. On the basis of the observations in the model system, a possible free radical mechanism for the antitumour action of the drug is suggested. PMID- 8225021 TI - Effect of the K+/H+ ionophore nigericin on response of A549 cells to photodynamic therapy and tert-butylhydroperoxide. AB - The K+/H+ ionophore nigericin dramatically increases killing of V79 cells and A549 cells by photodynamic therapy (PDT) sensitized by chloroaluminum phthalocyanine. Previous studies suggested that the interaction between PDT and nigericin is related to the ability of this ionophore to reduce intracellular pH (pHi). The present study was undertaken to test the possibility that nigericin, by lowering pHi, inhibits reductive detoxification of PDT-produced peroxides by enzymes of the glutathione (GSH) redox cycle and the pentose cycle. To test this possibility we examined the effects of nigericin on the toxicity and metabolism of a model peroxide, tert-butylhydroperoxide (tert-BOOH), in A549 cells, a cell line in which the GSH redox cycle is known to be the principal pathway for reduction and detoxification of tert-BOOH. We found that nigericin equilibrates pHi of A549 cells with extracellular pH (pHe) in a time-dependent manner. It increases the toxicity of tert-BOOH toward A549 cells, inhibits loss of tert-BOOH from the buffer overlying the cells, and reduces the rate of 14CO2 release from radiolabelled glucose, which is a measure of pentose cycle activity. These effects are significantly greater at pHe 6.40 than at 7.40. Monensin, a Na+/H+ ionophore which does not reduce pHi, does not enhance the toxicity of tert-BOOH and has only a minimal effect on tert-BOOH reduction. These data suggest that nigericin-induced inhibition of peroxide detoxification is at least a plausible mechanism by which the ionophore might interact with PDT. PMID- 8225022 TI - Xanthine oxidase activity in the circulation of rats following hemorrhagic shock. AB - Reactive oxygen metabolites generated from xanthine oxidase play an important role in the pathogenesis of ischemia-induced tissue injury. In a hemorrhagic shock model of ischemia-reperfusion, the intracellular enzyme xanthine oxidase was released into the vasculature. This intravascular source of superoxide (O2.-) and hydrogen peroxide (H2O2) interacted reversibly with glycosaminoglycans of vascular endothelium and markedly concentrated xanthine oxidase at cell surfaces, enhancing its ability to produce extensive damage to remote tissues. Rats were made hypotensive by hemorrhage, maintained for 2h, and reinfused with shed blood. Blood samples were obtained prior to hemorrhage and 15, 30, 60, and 90 min after reperfusion for determination of xanthine oxidase (XO), lactate dehydrogenase (LDH), and alanine transaminase (AST). These enzymes were not significantly elevated in control animals. Reperfusion after hemorrhage-induced ischemia resulted in significantly elevated AST and LDH in both low heparin (100 U/h) and high heparin (1000 U/h) groups. Xanthine oxidase was detected in the circulation only after 90 min reperfusion in the low heparin group and was elevated during the entire reperfusion period in the high heparin group. Studies with cultured vascular endothelium showed significant heparin-reversible binding of XO to cellular glycosaminoglycans. These results suggest that XO can gain access to the circulation following ischemia, where it then binds to the vascular endothelial cells to produce site-specific oxidant injury to organs remote from the site of XO release. PMID- 8225023 TI - Nitroxide radical biostability in skin. AB - Nitroxide radicals are important chemical tools in dermatologic research (e.g., for studying biophysical properties of skin lipids and epidermal membranes with the method of electron paramagnetic resonance, EPR, spectroscopy). However, nitroxides may loose their paramagnetic properties in biological tissues, which could limit their usefulness in biomedical applications. We analyzed the biostability of various chemical types of nitroxide radicals in keratinocytes, epidermis homogenate, and intact skin. EPR signal loss of imidazoline, pyrrolidine, piperidine, and oxazolidine nitroxides is attributed to their reduction to the corresponding hydroxylamine. The rate of nitroxide reduction in skin varies considerably with nitroxide ring structure and substitution. The order of nitroxide stability in isolated human keratinocytes, mouse epidermis homogenate, and intact mouse and human skin is imidazoline > pyrrolidine > di-t butylnitroxide (DTBN) > piperidine > oxazolidine. Cationic nitroxides are reduced much faster than neutral or anionic probes, presumably due to transmembrane electron shuttle or internalization. The results indicate that imidazoline- and pyrrolidine-type nitroxides should be used when high biostability of nitroxides is needed. Piperidine-type nitroxides are versatile probes for studying one electron transfer reactions in skin. PMID- 8225024 TI - Spin trapping of superoxide radicals following stimulation of neutrophils with fMLP is temperature dependent. AB - Oxygen radical formation by human neutrophils stimulated with a chemotactic peptide, formyl-methionyl-leucyl-phenylalanine (fMLP), was studied through the use of spin trapping and superoxide dismutase-inhibitable reduction of oxidized cytochrome c. Both methods provided comparable data on temperature-dependent kinetics of superoxide radical formation, but hydroxyl radicals were also detected in spin-trapping experiments. When superoxide generation was monitored at 37 degrees C, the respiratory burst lasted only a few minutes. If the neutrophils were stimulated at 37 degrees C, but superoxide measurements were done at room temperature, the respiratory burst was again transient. However, neutrophils persistently generated superoxide when both stimulation and subsequent measurements were performed at room temperature. In the presence of the actin polymerization inhibitor, cytochalasin B, superoxide generation was persistent, even when measurements were conducted at 37 degrees C. A possible explanation for these observations is that the fMLP receptor complexes quickly aggregate and are internalized at physiological temperature, but not at room temperature. Very little superoxide was formed if cells were kept at a temperature of 4 degrees C for 1 h prior to fMLP addition, which is consistent with decreased expression of the fMLP receptor at cold temperatures. PMID- 8225025 TI - The Fenton reagents. AB - Numerous transition metal ions and their complexes in their lower oxidation states (LmMn+) were found to have the oxidative features of the Fenton reagent, and, therefore, the mixtures of these metal compounds with H2O2 were named "Fenton-like" reagents. Using the Marcus theory and the experimental data in the literature, it is shown that in most cases the reaction of these metal complexes with H2O2 is unlikely to occur via an outer-sphere electron-transfer mechanism. It is suggested that the first step in this process is the formation of a transient complex LmM-H2O2n+, which may decompose to an .OH radical or a higher oxidation state of the metal, LmM(n + 2)+, or it may yield an organic free radical in the presence of organic substrates. Thus, the question whether free .OH radicals are being formed or not via the Fenton reaction depends on the relative rates of the decomposition reactions of the metal-peroxide complex and that of its reaction with organic substrates. Contradictory conclusions described from the study of different systems might only indicate that these relative rates are different in these systems. PMID- 8225026 TI - Luminol and lucigenin as detectors for O2.-. AB - Univalent oxidation of luminol and univalent reduction of lucigenin must precede reaction with O2.- if that reaction is to lead to luminescence. The assumption that luminol or lucigenin, per se, reacts with O2.- in a way leading to luminescence is incorrect, and leads to misinterpretation of results. The chemical reactions leading to the O2(.-)-dependent luminescences of luminol and of lucigenin are discussed. PMID- 8225027 TI - Kinetic properties of light-dependent ascorbic acid oxidation by melanin. AB - The kinetic properties of ascorbic acid oxidation by light-activated melanin granules demonstrate the presence of a specific reactive site on the melanin granule saturable by ascorbic acid. Increased light intensity increased the Vmax and reduced the Km of this reaction, indicating increased affinity of the active site for ascorbic acid. The kinetics of this reaction are not markedly changed in a reduced-oxygen environment. Ascorbic acid oxidation is competitively inhibited by isoascorbic acid, an epimer of ascorbic acid, while other tested reducing agents are inactive. The Ki for isoascorbic acid is 1 mM, about the same as the Km of ascorbic acid. PMID- 8225028 TI - Heat stress attenuates free radical release in the isolated perfused rat heart. AB - Prior heat stress leads to an enhancement of postischemic mechanical function and improvement in biochemical indices of injury in the rat heart, associated with an elevation in endogenous catalase activity. We have examined the effect of heat stress on free radical release during reperfusion in the isolated rat heart using electron spin resonance (ESR). Twenty four hours after heat stress or sham treatment, hearts were perfused in the Langendorff mode and subjected to 10 min of no-flow global ischemia followed by 10 min of reperfusion. Coronary effluent was collected at specific time points in PBN for ESR measurement. A PBN adduct was identified with characteristics consistent with an alkoxyl radical: PBN-LO. In sham hearts there was a rapid rise in adduct release to a maximum (228 +/- 15% of stabilization values, p < .05) occurring 1 min into reperfusion. In heat stress hearts there was no significant rise in adduct release during the reperfusion period. Pretreatment of hearts with 3-amino triazole, an inhibitor of catalase, failed to clarify whether the protection seen in heat stress hearts was a result of the elevation in catalase activity. These results suggest that heat stress protects the myocardium against the oxidative stress of ischemia reperfusion. PMID- 8225029 TI - 1st Annual Meeting of The Oxygen Society. Charleston, South Carolina, November 12 17, 1993. Abstracts. PMID- 8225030 TI - Induction of granulocytic maturation in HL-60 human leukemia cells by free radicals: a hypothesis of cell differentiation involving hydroxyl radicals. AB - Tumor cells usually contain lower superoxide dismutase (SOD) activity than differentiating cells, suggesting the involvement of oxygen free radicals in cell maturation. The effects of a system known to produce the OH. radicals were tested on HL-60 cells cultured under optimum conditions for 96 hr. Hydroxyl radicals were generated by a Fenton reaction, involving an ADP-Fe2+ (or ATP-Fe2+) complex and H2O2. Changes induced by OH. were compared to the effects of DMSO-induced differentiation of HL-60 cells. Cell numbers, viability, thymidine incorporation, TPA-induced NBT reduction and propidium iodide staining in flow cytometry were determined. The OH. generating system inhibited the growth and thymidine incorporation of leukemic cells in a manner dependent on the dose of added H2O2 (from 0.005 to 0.05 mM). In addition, an increasing proportion of the treated cells displayed signs of cell differentiation. In DMSO-treated cells, SOD and catalase activities increased after 6 days of culturing. The results show that a portion of the OH. free radicals derived from H2O2, produced by the action of SOD, may be a necessary prerequisite for differentiation, whereas an overproduction of OH. causes cell lethality or aging. We suggest that OH. free radicals may have a more complex role in cell physiology than simply causing oxidative damage. PMID- 8225031 TI - Reaction kinetics of alpha-tocopheroxyl radical with biologically and pharmacologically active substances. AB - The alpha-tocopheroxyl radical (alpha TR.)generated in the reaction with 1,1 diphenyl-2-picrylhydrazyl in n-butanol decayed according to second-order kinetics with a rate constant k alpha = 3 x 10(3) M-1s-1 as determined by EPR spectroscopy. Various biologically and pharmacologically active substances like isoprenaline (ISO), epinephrine (EPI), histamine (HIS), stobadine (STO), nafazatrom (NAF) and Kampo C medicine (KMC) accelerated the decay rate of alpha TR(.). The whole process is formally a third-order reaction with the rate constants (in 10(9) M-2s-1): ks(ISO) = 1.28, ks(NAF) = 1.25, ks(EPI) = 0.6, ks(HIS) = 0.4, and ks(STO) = 0.1. In the kinetics of the reaction mechanism, bimolecular intermediates are assumed and the rate constants of their formation were determined. PMID- 8225032 TI - Purification of Cu, Zn-superoxide dismutase isoenzymes from fish liver: appearance of new isoforms as a consequence of pollution. AB - Liver cell-free extracts of fish (Mugil sp.) from polluted environments show new Cu,Zn-SOD isoenzymes when analyzed by polyacrylamide gel electrophoresis or isoelectrofocusing followed by in situ staining for SOD activity. The most active isoenzymes, with pI 6.1 and 5.1, were present both in control and problem samples while the isoenzymes of intermediate pI value showed significant differences. Fish from control areas showed three intermediate isoenzymes with pI 5.7, 5.5 and 5.4 (the last one quite faint) while polluted animals showed three bands of pI 5.9, 5.45 and 5.35, this last very intense. To further characterize their utility as biomarkers, Cu,Zn-SOD isoenzymes from polluted fish livers were purified to homogeneity. Five superoxide dismutase peaks were purified, named thereafter I (pI 6.1) to V (pI 5.1) respectively. Isoenzymes I and V displayed the highest specific activity. Upon incubation with moderate H2O2 concentrations, pure isoenzyme I yielded more acidic bands with pI 5.5, 5.45 and 5.35, this last being predominant. The pure isoenzyme V generated only a new band of pI 5.0. Concomitant with oxidation, the activity of peaks I and V was lost in a H2O2 concentration-dependent manner. The pattern of the new acidic bands generated upon the oxidixing treatment of isoenzyme I closely resembles that observed in crude extracts from polluted animals. PMID- 8225033 TI - In vivo 31P NMR spectroscopy studies of halothane induced porcine stress syndrome. No effect of C-phenyl N-tert-butyl nitrone (PBN). AB - Porcine stress syndrome (PSS) which is an example of malignant hyperthermia (MH) in swine has previously been attributed to oxidative stress primarily due to an inherited antioxidant abnormality in MH susceptible (MHS) animals. C-phenyl-N tert-butyl nitrone (PBN), a free radical spin trap, was selected to investigate whether free radicals are involved in MH. If free radicals cause the MH stress attack, then PBN should alter the time required for the onset of the stress attack, or perhaps protect the animal from experiencing the stress attack. In vivo phosphorus-31 (31P) magnetic resonance spectroscopy (MRS) was used to monitor metabolism in three to four week old normal and MHS piglets administered halothane as the stress challenge. Malignant hyperthermia was not reproducibly induced by halothane anesthesia. For those animals which did develop MH a dramatic fall in the level of PCr and a rise in the level of Pi was detected by 31P MRS. Intravenous administration of PBN prior to halothane exposure had no effect on the number of animals experiencing the stress attack. PBN does not appear to prevent, delay or reverse the onset of halothane-induced MH in three to four week old MHS piglets. The primary events leading to the MH syndrome do not appear to be influenced by the intervention of the type of free radicals normally trapped by PBN. PMID- 8225034 TI - Lipid peroxidation in hyperlipidaemic patients. A study of plasma using an HPLC based thiobarbituric acid test. AB - The question of "increased lipid peroxidation" in plasma from hyperlipidaemic patients was investigated using an improved HPLC-based assay for thiobarbituric acid-reactive material. Levels of TBARS in healthy human controls were at or close to zero, provided that butylated hydroxytoluene was added to the sample with the TBA reagents. Levels of plasma TBARS in hyperlipidaemic patients were elevated, although the absolute levels were much lower than those reported previously in the literature. PMID- 8225035 TI - Are free radicals involved in the pathobiology of human essential hypertension? AB - Possible involvement of reactive oxygen species and nitric oxide in the pathogenesis of human essential hypertension was investigated. It was observed that both superoxide anion and hydrogen peroxide production by polymorphonuclear leukocytes and the plasma levels of lipid peroxides are higher in uncontrolled essential hypertension compared with normal controls. Nitric oxide levels measured as its stable metabolite nitrite, as an index of nitric oxide synthesis, revealed its levels to be low in hypertensive patients. Superoxide anion, hydrogen peroxide, lipid peroxides and nitric oxide levels reverted to normal values after the control of hypertension by drugs. The concentrations of anti oxidants such as vitamin E and superoxide dismutase were found to be decreased in patients with uncontrolled hypertension. Several anti-hypertensive drugs inhibited lipid peroxidation in vitro. Angiotensin-II, a potent vasoconstrictor, stimulated free radical generation in normal leukocytes which could be blocked by calmodulin antagonists. These results suggest that an increase in free radical generation and a simultaneous decrease in the production of nitric oxide and anti oxidants such as SOD and vitamin E occurs in essential hypertension. This increase in free radical generation can inactivate prostacyclin and nitric oxide and decrease their half life which can lead to an increase in peripheral vascular resistance and hypertension. PMID- 8225036 TI - Spin trapping of superoxide from glass adherent polymorphonuclear leukocytes induced by N-formylmethionyl-leucyl-phenylalanine. AB - Dahinden et al. reported that N-formylmethionyl-leucyl-phenylalanine (fMLP) induced superoxide release from polymorphonuclear leukocytes (PMNs) lasted more than 60 min when the cells were allowed to attach to a petri dish before induction. In contrast, it lasted only for 2.5 min when cells were in suspension (J. Clin. Invest. 72: 113-121, 1983). In spite of this report, the effect of cell adhesion has been ignored in most spin trapping studies of superoxide release from PMNs. This study shows that most PMNs in a quartz flat electron paramagnetic resonance (EPR) cuvette which was placed horizontally adhered to the wall within 3 min. In contrast, if the cuvette was placed vertically, only 20-30% of the cells became adherent in 30 min. We performed spin trapping studies using 5,5 dimethylpyrroline-N-oxide (DMPO) as a spin trap, and monitored the effect of cell adhesion on superoxide generation. When spin trapping was conducted on PMNs in suspension, the EPR signal of superoxide adduct (DMPO-OOH) was undetectable after stimulation with fMLP. However, PMNs which were allowed to adhere to the cuvette after stimulation generated superoxide for hours. Moreover, when PMNs were allowed to adhere prior to the stimulation, the magnitude of superoxide release was augmented three-to fourfold. Unlike fMLP, phorbol myristate acetate (PMA), which has been most commonly used in spin trapping studies, induced superoxide release which was not influenced by cell adhesion. We emphasize the importance of specifying the cell-adhesion-state in spin trapping studies. PMID- 8225037 TI - Modulation of glutathione and glutathione dependent antioxidant enzymes in mouse heart following doxorubicin therapy. AB - The toxicity of the antineoplastic agent doxorubicin (DOX) has been shown to be moderated by the antioxidant enzyme glutathione peroxidase. It has been reported that acute doses of DOX can cause an inhibition of glutathione peroxidase in cardiac tissue, that may render this tissue especially susceptible to further prooxidant damage. In this study, multiple DOX treatments at a therapeutic dose were assessed for their effect on the antioxidant enzyme status of cardiac and kidney tissue. DOX was administered i.p. (5 mg/kg) once a week for two weeks to male balb/c mice. The activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPOX) and glutathione reductase (GR) were measured 1, 2 and 7 days following the second DOX treatment in both heart and kidney. Levels of reduced glutathione (GSH) were also measured in cardiac tissue at these same times. Cardiac levels of GPOX and GR showed a time dependent decrease in activity, with 10% and 12% inhibition for GPOX and GR, respectively, at 7 days post second treatment. Cardiac levels of GSH also showed a significant decrease, approximately 15%, at 7 days post second treatment. Cardiac levels of SOD and CAT as well as kidney levels of all four antioxidant enzymes were not affected by DOX treatment. These data suggest that DOX given in a therapeutic regimen, at a therapeutic dose, can cause decreases in cardiac levels of GPOX, GR and GSH that could render the heart especially susceptible to further oxidative challenge. PMID- 8225038 TI - Interaction of ethanol with haemoglobin: implications for alcohol abuse. AB - Ingestion of large amounts of ethanol results in a variety of toxicological consequences of which reduced oxygen supply is a marked respiratory effect. Experimentation involving the in-vivo administration of ethanol to rats, identifies the formation of ferri-haemoprotein species and hydroxyl radicals in the liver. Our work on the in-vitro effect of ethanol on oxyhaemoglobin shows the formation of ferri-haemoglobin and the alpha-hydroxyethyl radical as an intermediate. The impairment of oxygen transport efficiency of haemoglobin may be a result of its oxidation to the non-functional ferrihaemoglobin form. PMID- 8225039 TI - Enhanced enzymatic degradation of radical damaged mitochondrial membrane components. AB - The location of a protein (soluble or membrane-bound) influences the extent of oxidative damage caused by free radicals. It has been established that after radical attack, soluble proteins can become more susceptible to hydrolysis by individual proteinases than native proteins. We have now examined the hydrolytic susceptibility following radical attack of a protein that is located within a membrane environment, mitochondrial monoamine oxidase (MAO). After exposure to oxygen radicals generated by gamma irradiation, hydrolysis of sub-mitochondrial particles (SMP) containing MAO was increased in three respects. First, the generation of small fragments of MAO by the proteinases elastase and trypsin, was enhanced. Second, the generation by these enzymes and by phospholipase A2 of non sedimentable membrane fragments containing MAO was also increased. Third, autolysis of SMP was enhanced. Hence, proteins located within membranes may become more susceptible to enzymatic degradation following oxidative damage. PMID- 8225040 TI - Hyperbaric oxygen therapy increases free radical levels in the blood of humans. AB - It has been postulated that exposure to high concentrations of oxygen results in increased oxygen radical production which may account for the toxic effects of excessive exposure to oxygen. Examination of blood from persons undergoing hyperbaric oxygen (HBO) exposure, by low temperature electron spin resonance (ESR) spectroscopy, demonstrated a marked increase in the magnitude of a signal with properties consistent with a free radical (g = 2.006). The signal diminished to baseline levels within 10 minutes of cessation of HBO exposure. Further in vitro studies of blood revealed an ESR signal generated in red blood cells by oxygen, and dependent on oxyhaemoglobin, which had characteristics indistinguishable from those of the ESR signal of ascorbate radical and the signal in blood from persons undergoing HBO exposure. It is postulated that HBO exposure increases ascorbate radical levels in blood, which is likely to reflect increased ascorbate turnover in human red blood cells. PMID- 8225041 TI - The role of glutathione in protection against DNA damage induced by rifamycin SV and copper(II) ions. AB - Incubation of calf thymus DNA in the presence of rifamycin SV induces a decrease in the absorbance of DNA at 260 nm. The effect, was found to be proportional to the antibiotic concentration and enhanced by copper(II) ions. In the presence of rifamycin SV and copper(II), a significant increase in thiobarbituric acid reactive (TBA-reactive) material is also observed. This effect is inhibited to different degrees by the following antioxidants: catalase 77%; thiourea 72%; glutathione (GSH) 62%; ethanol 52%; and DMSO 34%, suggesting that both hydrogen peroxide (H2O2) and hydroxyl radicals (OH.) are involved in DNA damage. Rifamycin SV-copper(II) mixtures were also found to induce the production of peroxidation material from deoxyribose and, in this case, glutathione and ethanol were the most effective antioxidant substrates with inhibition rates of 91% and 88% respectively. Electrophoretic studies show that calf thymus DNA becomes damaged after 20 min. incubation in the presence of both agents together and that the damaged fragments run with migration rates similar to those obtained by the metal chelating agent 1,10-phenanthroline. Normal DNA electrophoretic pattern was found to be preserved by catalase, and GSH at physiological concentrations and by thiourea. No protection is observed in the presence of ethanol or DMSO. The results obtained indicate the involvement of different reactive species in the degradation process of DNA due to rifamycin SV-copper(II) complex and emphasize the role of reduced glutathione as an oxygen free radical scavenger. PMID- 8225042 TI - Elevation of the level of thiobarbituric acid-reactive products in hindleg skeletal muscle of dystrophic mice, but non-elevation in tongue muscle. AB - In order to understand the pathogenesis of mouse muscular dystrophy, we investigated the levels of the thiobarbituric acid-reactive substances (TBARS), H2O2 and NADPH oxidase activity, which were relative to the acceleration of oxidative conditions, in tongue and hindleg skeletal muscles from C57BL/6J-dy mice. The TBARS content (702 nmol/g protein) in skeletal muscles from 2-months old dystrophic mice was increased significantly over that (384 nmol/g protein) in muscles from age-matched normal mice. The H2O2 concentration in dystrophic skeletal muscles was 30% higher than that in normal ones. Microsomal NADPH oxidase activity which was related to the production of superoxide anions, was similar between dystrophic muscles (4.66 nmol/10 min/mg protein) and normal muscles (4.11 nmol/10 min/mg protein). These results indicate that oxidation is accelerated in the dystrophic muscles. However, the TBARS content in the tongues of dystrophic mice was identical to that of normal mice. This finding supports our bone-muscle growth imbalance hypothesis for the pathogenesis of mouse muscular dystrophy. PMID- 8225043 TI - Premature rupture of the membranes--aggressive versus conservative approach: effect of tocolytic and antibiotic therapy. AB - The purpose of this randomized, prospective study was to evaluate the efficacy of tocolytic and antibiotic therapy in the prolongation of pregnancy and neonatal outcome in the treatment of premature rupture of the membranes without clinical labor. Delivery was delayed for 48 h, 7 days and beyond 35 weeks of gestation in 87, 39 and 18%, respectively, of patients in the treated group (n = 39) compared with 50, 12 and 17% of patients in the nontreated group (n = 42). The incidence of a low Apgar score (< 7 at 5 min), requiring artificial ventilation, and infectious morbidity was more common in the treated group than in the nontreated group (18 vs. 0, 41 vs. 17 and 39 vs. 17%, respectively). There was no significant cost difference in survivors between the treated and nontreated groups, although the mothers in the treated group were significantly more expensive. From these observations, it appears that tocolysis and antibiotics are not effective in PROM cases. PMID- 8225044 TI - Lipid profiles in healthy fertile Singaporean women. AB - Serum lipid profiles were studied in 167 healthy fertile Singaporean women, aged between 18 and 40 and comprising 114 Chinese, 28 Malays and 25 Indians. Parity or ethnic differences did not affect lipid concentrations. Except for triglycerides which showed a decreasing trend, there was no significant variation in lipid concentrations with age. However, all lipid concentrations except HDL cholesterol (which decreased) appeared to increase with body mass index. Compared with other populations, these Singaporean women appeared to have higher mean concentrations of total cholesterol and lower mean concentrations of HDL cholesterol. The possibility that these differences could have contributed to the increasing incidence of coronary heart disease in Singapore is discussed. PMID- 8225045 TI - Bone loss in young women with primary hypogonadism and its prevention with calcitonin. AB - A total of 63 healthy young women with primary amenorrhea, 37 control women at the same age with normal ovarian function and 118 postmenopausal women underwent bone mineral density (BMD) measurements at the lumbar spine (L2-L4) by dual photon absorptiometry. Mean +/- SE BMD value in the primary amenorrheic patients (0.878 +/- 0.015 g/cm2) was significantly lower than in both the control ovulatory women (1.069 +/- 0.026 g/cm2) and the postmenopausal patients (0.967 +/ 0.013 g/cm2) (p < 0.05). When hormonal parameters were compared between primary amenorrheic and ovulatory control cases, only the prolactin mean levels were found to be significantly lower in the first group (8.74 +/- 0.86 vs. 16.31 +/- 1.05 ng/ml; p < 0.001). Serum total alkaline phosphatase activity and LDL cholesterol were negatively correlated with bone mineral density. Total cholesterol and lipid levels were significantly higher in women with primary amenorrhea when compared with the eumenorrheic control group (p < 0.001). PMID- 8225046 TI - Acute tissue effects during transcervical endometrial resection. AB - The acute tissue effects of transcervical endometrial resection (TCRE) with a standard Iglesias resectoscope using glycine 1.5% for irrigation were studied in 8 women prior to hysterectomy. Combinations of 80 or 120 W cutting current with blend 1 or 2 were applied for endometrial resection, preceded by cornual endometrial coagulation with the roller ball electrode with a coagulation effect of 40 or 60 W. The temperature was measured at the uterine vessels, ovarian ligaments and serosal surface. The increase in temperature at the serosal surface was 2.0 degrees C during cornual coagulation and 0.3 degrees C during endometrial resection, independent of the current effect applied. The maximum depth of tissue damage was 1.7 mm. No change in temperature was found at the uterine vessels or ovarian ligaments. The tissue destruction and the increase in temperature of the uterine surface are minimal, and TCRE offers excellent histological material. Careful coagulation/resection in the cornual and isthmus regions is recommended. PMID- 8225047 TI - Uterine rupture during a trial of labor in a case with a unicornuate uterus and a previous cesarean section. AB - We describe a case of uterine rupture during labor in a patient with a previous cesarean section and a unicornuate uterus. The case may encourage obstetricians to carry out elective repeated cesarean sections in cases with previous cesarean sections and uterine anomalies. PMID- 8225048 TI - Death of one twin followed by extremely variable flow velocity waveforms in the surviving fetus. AB - Intrauterine death of one fetus after the second trimester in a twin pregnancy, with continuation of the pregnancy is a rare complication. The risks of morbidity and mortality for the surviving fetus are high. A 32-year-old woman was admitted to the antenatal ward at 27 weeks gestation because of intrauterine death of one twin. During the first 24 h after the death of one twin, Doppler ultrasound assessment showed a remarkable variability in flow velocity waveforms in the umbilical artery of the surviving fetus. Changes from reversed to normal end diastolic flow velocities were recorded within 6 min. These findings are explained by twin-to-twin transfusion due to intravascular blood pressure changes, or by release of vasoactive substances by the dead fetus. PMID- 8225049 TI - Response of blood flow velocity waveforms in the abdominal aorta and cotyledonary artery to regional embolization of the umbilical circulation in fetal sheep. AB - Regional embolization of the placental circulation was performed in 3 anesthetized fetal sheep by retrograde administration of microspheres in a cotyledonary branch of one of the umbilical arteries. Registrations were made of blood velocity waveforms in the fetal abdominal aorta and in a major cotyledonary artery supplying the embolized placental region. Among the fetal lambs examined, a corresponding degree of regression was found between the decrease in umbilical blood flow and increase in the pulsatility index of the abdominal aorta waveforms (coefficient of variation: 0.19). In contrast, the degree of regression between the decrease in umbilical blood flow and increase in the pulsatility index of the cotyledonary artery waveforms varied per fetus (coefficient of variation: 0.79). It is suggested that in human fetuses, in the case of discordant umbilical artery blood flow velocity waveforms, the best information on fetoplacental circulatory competence is provided by the aorta blood velocity waveform. PMID- 8225050 TI - Chromosome breakage in individuals with single-cell structural aberrations and habitual abortions. AB - The rate of spontaneous and methotrexate (MTX)-induced chromosome breakage was studied in individuals with a history of habitual abortions in whom a structural chromosomal aberration was found in a single cell during routine cytogenetic analysis. Twelve such individuals were selected because they were not under the influence of any known mutagenic factor such as smoking, alcohol, medication and apparent irradiation; they were compared to 12 age- and sex-matched control parents. A detailed statistical analysis revealed that the spontaneous and MTX induced chromosome breakage was significantly increased in the abortion group. The MTX-induced breakage rate was especially elevated in the women of the abortion group. PMID- 8225051 TI - Alpha-adrenergic receptors in human myometrium: changes during pregnancy. AB - Myometrial biopsies from 80 women, delivered by cesarean section in 26-42 completed weeks of pregnancy, were analyzed using radiolabeled alpha 1-antagonist [3H]-prazosin and alpha 2-antagonist [3H]-rauwolscine. Statistical analysis was performed using Student's t test (two-tailed), where p < 0.05 was considered significant. The number of alpha 2-adrenergic receptors did not vary (mean value weeks 37-39 = 17.2 +/- 1.3 fmol/mg protein), while the dissociation constant (Kd) for alpha 2-receptors tended to increase with increasing gestational age, implying reduced affinity near term (p < 0.05, mean values 3.4 +/- 0.2 nM). alpha 1-Receptors, however, increased significantly (p < 0.01) in the pregnancy weeks 37-39 (mean value 29.2 +/- 2.9 fmol/mg prot) while Kd was unaltered. Increased number of alpha 1-receptors, along with a decreased affinity of alpha 2 receptors, could, in combination with other factors, play a role in the initiation of labor. PMID- 8225052 TI - Is oxytocin involved in the deterioration of glucose tolerance in gestational diabetes? AB - The present study assessed the possible role of oxytocin in the deterioration of glucose tolerance in gestational diabetes. Plasma levels of oxytocin, insulin, glucagon and glucose were measured at the time of a 400-kcal breakfast meal tolerance test in 12 women with gestational diabetes and 12 normal pregnant women in the third trimester. The gestational diabetic women had higher basal levels of insulin and an enhanced, delayed and prolonged insulin response to the breakfast. The same differences occurred in the glucose levels. There was no significant difference in the glucagon levels between the two groups. In the normal pregnant women, a significant (p < 0.05) though small rise in glucagon levels occurred 30 min after the ingestion of the breakfast. Oxytocin levels were not affected by the breakfast, and there was no clear difference between the two groups. The metabolic differences between the normal pregnant and gestational diabetic women were not related to any differences in oxytocin levels. In conclusion, we found no evidence of a role of oxytocin in the alteration of glucose metabolism in women with gestational diabetes. However, since alterations in oxytocin levels of possible significance for an impaired glucose tolerance are found in type 1 diabetic and extremely obese patients, further studies are needed in women with gestational or manifest diabetes. PMID- 8225053 TI - Late vaginal induced abortion after a previous cesarean birth: potential for uterine rupture. AB - This descriptive study was conducted to evaluate the risk of uterine rupture in cases of late vaginal induced abortions among women with previous cesarean sections. 23 women were referred at a mean gestational age of 23.9 (SD 6.9) weeks of gestation after one or two cesarean sections. Indications for terminating the pregnancy were maternal diseases in 4 cases and fetal anomalies in 19 cases. RU 486 and/or prostaglandins were used for cervical ripening and to induce labor. Vaginal birth was obtained in 20 cases (86.9%) with an average duration of 72 (SD 52) h. Cesarean section was performed in the remaining 3 women because no cervical dilation could be obtained. One uterine rupture occurred and was treated with conservation of the uterus. Late termination of pregnancy in such cases can be achieved without cesarean section with a high success rate. PMID- 8225054 TI - Specific IgM antibodies as parameters of toxoplasma infection in pregnancy. AB - The aim of this investigation was to determine the time of toxoplasma infection more precisely by using an indirect fluorescent antibody test and indirect hemagglutination test in 51 pregnant women whose Sabin-Feldman test and immunosorbent agglutination assay were positive. Analysis of the results of sera tested with the two additional serological assays showed that the infection was acquired during pregnancy in 19 cases, while the other pregnant women had become infected before conception. PMID- 8225055 TI - Group I and group II phospholipase A2 in serum during normal and pathological pregnancy. AB - Phospholipase A2 groups I (pancreatic) and II (synovial) could be a link between local and systemic changes in pregnancy, reflected in catalytic activity. We studied whether normal pregnancy, preeclampsia, preterm labor and four other diseases have processes involving serum phospholipase A2s. Pancreatic and synovial-type phospholipase A2 were measured in the serum of 59 normal pregnant women and 89 patients with pathological pregnancy by newly developed time resolved fluoroimmunoassays, and the catalytic activity by a radiochemical method using micellar phosphatidylcholine as substrate. During pregnancy weeks 6-14, synovial-type phospholipase A2 and catalytic activity were elevated 2- to 4-fold, but at 37 weeks values were normal. Pregnancy-induced hypertensive diseases increased by 4- to 10-fold the concentration of synovial-type phospholipase A2, reflected in catalytic activity. In 8 out of 14 cases, the enzyme was increased if the fetus was to be delivered prematurely. The enzymes studied remained within the reference interval in cases of hepatogestosis, fetal asphyxia, diabetes and twin pregnancy. Newly developed specific immunoassays for measuring different types of phospholipase A2 in serum can provide insights for clinical follow-up. PMID- 8225056 TI - [Excimer laser photorefractive keratectomy for myopia]. AB - We have used excimer laser during the past year to correct refractive errors (myopia and hypermetropia) and to remove superficial corneal scars. 714 eyes underwent excimer laser photorefractive keratectomy for correction of myopia and 81% of the patients achieved uncorrected visual acuity of 6/12 or better without glasses. 17 patients underwent a second treatment to correct residual myopia and all of them had uncorrected visual acuity of 6/12 or better 6 months later. The results show that excimer laser photorefractive keratectomy is a safe and efficient procedure to correct myopia. The possibility of various postoperative complications appearing over the long term suggests cautious optimism. PMID- 8225057 TI - [Thrombolysis for evolving myocardial infarction in a rural primary care clinic]. AB - Early thrombolytic therapy gives maximum benefit in acute myocardial infarction. In remote rural areas with no mobile intensive care service there is a significant delay between onset of symptoms and administration of thrombolytic therapy which has a critical impact on revascularization. Thrombolytic therapy with streptokinase 1,500,000 U was given in a primary care rural clinic to 2 patients with evolving myocardial infarction 45-50 minutes from onset of symptoms. In both patients, who were transported to hospital after the therapy, there were clinical signs of reperfusion. There were no complications during treatment or transportation. We conclude that thrombolytic therapy given for evolving myocardial infarction in a rural primary care clinic is possible and safe. PMID- 8225058 TI - [Do exercise-induced chest pain and left bundle branch block always indicate coronary artery disease?]. AB - The development of intermittent left bundle branch block (LBBB) and of chest pain on exercise stress testing, usually considered indicative of coronary artery disease, does not always indicate occlusive coronary artery disease (OCAD). In this specific entity, neither the echocardiogram nor radionuclide angiography is accurate in predicting true OCAD. 2 of 6 patients with rate-dependent LBBB and chest pain on exertion but without angiographic evidence of OCAD are reported. Based on these cases and those in the literature, we suggest characteristics of chest pain or discomfort, and different parameters of exercise testing, to help identify patients without OCAD. Exercise stress test electrocardiography is an important tool in diagnosing this unique entity. PMID- 8225059 TI - [Urinary tract infections in the community]. AB - 4,612 urine samples were sent to this laboratory in 1990, and the results of those sent during a single week in each of the 4 seasons were studied. Of 4501 urinary cultures sent from the general population, 811 (80%) were positive for E. coli, Klebsiella and Proteus mirabilis. 60% were resistant to ampicillin, 35% to cotrimoxazole and 20% to cephalexin. Of 111 urine samples sent from senior citizen homes, 57 (80%) were positive for E. coli, Proteus mirabilis, Klebsiella and Pseudomonas. 74% were resistant to ampicillin, 72% to cotrimoxazole and 46% to cephalexin. Ofloxacin and norfloxacin were very effective in urinary tract infections in the general population, with 94% and 97% sensitive to them, respectively. 68% of the urinary cultures sent from the senior citizen centers were sensitive to each of them. PMID- 8225060 TI - [Stages in the licensing process for immigrant physicians in Israel]. AB - The latest wave of immigration, mostly from the former Soviet Union, has brought with it a large number of physicians. They are required to go through various stages of a licensing process in order to practice medicine in Israel. This overview describes the process, from arrival in Israel to the issuing of the license to practice medicine. From September 1989 to December 1991 some 9,800 of arriving immigrants identified themselves as physicians to Ministry of Absorption officials. However, only about 7,000 submitted requests for a medical license to the Ministry of Health. Physicians with at least 20 years of experience need not take an examination, but are required to work under supervision for 6 months in order to receive a license. Of those requesting a license, 1/3 had 20 years or more of professional experience, while the others had to take licensing examinations. By the end of December 1991, some 2,900 physicians had received licenses to practice medicine. This indicates that many immigrant physicians have yet to complete the licensing process. PMID- 8225061 TI - [Conservative surgery in penetrating renal injuries]. AB - In 11 of 21 patients with penetrating renal injury operated on during a year, salvage of the involved kidney or part of it was successful. We conclude that accurate diagnosis and staging of penetrating renal trauma, when combined with a conservative surgical approach, allows salvage of functional renal parenchyma in a large proportion of cases. PMID- 8225062 TI - [Primary breast lymphoma]. AB - A 66-year-old woman with a previous diagnosis of benign histiocytosis of the right breast developed intermediate grade malignant lymphoma of her left breast 6 years later. It is suggested that the primary disease was actually a low-grade lymphoma of the MALT subgroup (mucosa-associated lymphoid tumors) which 6 years later had undergone transformation into high-grade lymphoma. This assumption is based on the known histological similarity between MALT lymphomas and benign histiocytosis. PMID- 8225063 TI - [Continuity of care and hospital admission rates in a rural family practice]. AB - Hospitalization data may be used as an indicator of the quality of care in family practice. There were marked differences between hospitalization rates in a rural family practice and the corresponding national rural rates, which raised questions about the reasons for these differences. All 309 hospital admissions recorded between 1987 and 1990 in a rural family practice were then studied to determine the hospital admission pattern. The crude hospitalization rate of this practice over the period studied was 74/1000 person-years at risk. After indirect standardization with the national Jewish rural hospitalization rate, the actual number of hospitalizations for the family practice (309) was 24% less than would have been expected (407) had the national rural rate applied to the family practice population during the study period. The decreases between actual and expected admissions were greater in the infant, adolescent and elderly age groups. Continuity-of-care by a single provider in the study practice is presented as an important factor in determining the smaller number of practice hospitalizations recorded. PMID- 8225064 TI - [Future directions in family medicine and residency curriculum for the 21 century]. PMID- 8225065 TI - [Quality assurance in primary care]. PMID- 8225066 TI - [Intravascular ultrasound: a new imaging technic and its clinical application]. PMID- 8225067 TI - [Chickenpox and pregnancy]. PMID- 8225068 TI - [Excimer laser correction of myopia]. AB - Excimer laser photorefractive keratectomy was performed on 100 myopic eyes. The preoperative spherical equivalent of myopia ranged from -2.00 to -9.99 diopters (D), mean -4.80 +/- 2.20 D. The procedure was performed under local anesthesia using 0.4% topical benoxinate HCl; corticosteroid eye drops were then used for up to 3 months. All patients achieved corrected visual acuity within 1 line of corrected preoperative visual acuity. The only complication was subepithelial reticular haze seen in all patients, which peaked at 4 weeks after the procedure and gradually diminished within 3 months. After follow-up of from 6-11 months, 93% of patients had achieved uncorrected visual acuity of 6/12 or better. Mean postoperative refraction was -0.20 +/- 0.97 D. 69% achieved a correction within +/- 0.50 diopter of emmetropia, 85% within +/- 1.00 D and 96% achieved corrected visual acuity of within 1 line of preoperative corrected visual acuity. This study and those in the literature show excimer laser to be a promising surgical treatment for myopia. PMID- 8225069 TI - [Psychiatric aspects of steroid therapy]. PMID- 8225071 TI - [The use of excimer laser in refractive surgery]. PMID- 8225070 TI - [TAPS Research Project: study of the closure of two mental hospitals in London, and reprovision in the community for their populations. Part A: Objectives, design, instruments]. PMID- 8225072 TI - [Pertussis--why to vaccinate?]. PMID- 8225073 TI - [New methods for predicting sperm fertilizing ability]. AB - There is no reliable method for screening for the fertilizing ability of sperm. 3 new methods are presented: the freezing test (FT), hypoosmotic swelling test (HOST) and migration-sedimentation test (MST). The first 2, test the functional integrity and stability of the sperm membrane under certain osmotic conditions. The MST determines ability to separate high quality sperm. All tests were performed using either fresh semen of husbands, or thawed semen of donors used for fertilization in the IVF unit. Results of the FT were not correlated with the fertilizing ability of sperm in IVF, but HOST results with fresh, but not thawed semen, were correlated. Fertilization of oocytes in IVF was significantly better when HOST value of fresh semen was 45% or more. Motility and morphological normality of sperm separated by MST were better in ejaculates that fertilized ova under in vitro conditions, than in those that did not. Sperm separation by MST of 25% or more of motile sperm, or of more than 1.5 million motile sperm in the sample, indicated statistically significant ability for oocyte fertilization in an IVF system. PMID- 8225074 TI - [TAPS Research Project: study of the closure of two mental hospitals in London and providing in the community for their populations. Part B: Interim results]. PMID- 8225075 TI - [Desmopressing (DDAVP) treatment in patients with bleeding tendency]. PMID- 8225076 TI - [Guidelines for coronary revascularization. Ad Hoc Committee for Guidelines for Coronary Revascularization]. PMID- 8225077 TI - [Molecular biology in the diagnosis of infectious diseases]. PMID- 8225079 TI - [Are our babies becoming larger?]. PMID- 8225078 TI - [Prevention of heroin reabuse--value of naltrexone]. PMID- 8225080 TI - [Epidemiology of Hansen's disease in Israel]. AB - There are currently 200 patients with Hansen's disease in Israel who are being followed by the Hansen's Disease Government Hospital and the Ministry of Health (prevalence 4.4/100,000). Most of them immigrated from countries where the disease is endemic. Dermatological findings dominated the initial clinical picture, although 5% of patients are asymptomatic contacts of known cases. Age at onset of disease was less than 20 years in 1/4 of the cases. The incidence in Israel is falling: 0.4/100,000 in 1985-89 compared to 3.6/100,000 in 1950-54. Neurologic and dermatologic findings in an immigrant of any age originating from countries where Hansen's disease is endemic, should prompt appropriate diagnostic evaluation, even years after immigration to Israel. Contacts of known cases of Hansen's disease should be aggressively screened, even if asymptomatic. PMID- 8225081 TI - [Psychiatric hospitalization trends and predicted needs]. AB - This study attempts to predict the need up to 1995 for psychiatric beds in Israel, based on the trends of psychiatric hospitalization during 1986-1990. Additional need created by the recent large waves of immigrants to Israel are taken into account. Analysis was separate for chronic in-patients, (continuous stay of 1 year or more) and for acute in-patients (stay of less than 1 year). The data were drawn from the National Psychiatric Case Register. Among acute in patients, the admission rate of immigrants was higher than that of the veteran population, thus counterbalancing the trend to a decrease in admissions among the latter. It is assumed therefore that the present rate of 0.4 acute patients/1000 population will not change. However, an additional 300 new beds will be needed due to increase in the general population. With regard to chronic in-patients, a continuous decrease of about 50 chronic in-patients per year has been observed. Assuming 700,000 new immigrants will have arrived in Israel between 1990-1995, and that the proportion of patients admitted who remain in continuous hospitalization is larger among new immigrants than among the veteran Israeli population (10% vs. 6%), the beds expected to be vacated by chronic patients will be filled by new immigrant patients. The expected rate of chronic in-patients will nevertheless decrease from the present rate of 1.0/1000. to 0.8/1000 by 1995. PMID- 8225082 TI - [Endometriosis of the bladder]. AB - We report a case of endometriosis of the bladder in a 32-year-old healthy woman suffering from longstanding suprapubic pain, and urinary urgency and frequency. The disease, which is uncommon, usually has a high morbidity rate and is difficult to diagnose. A high level of suspicion is necessary for diagnosis, since neither the clinical presentation, nor imaging procedures are pathognomonic for endometriosis. PMID- 8225083 TI - [Masked mastoiditis]. AB - Masked mastoiditis defines a subclinical infectious inflammatory process of the mucosal lining and the bony structure of the mastoid air cells, with intact tympanic membrane. It follows an apparently well-treated recent acute otitis media. Because of obstruction of the tympanic diaphragm by mucosal swelling, polypoid mucosa or granulation tissue, a complete separation of the tubo-tympanic cavity from the mastoid air cell system results. The latter remains unaereated and the local infectious mucosal disease progresses to osteitis. Probably due to colonizing flora in this unventilated media, the developing bone infection is low grade, without pus formation. The clinical features of the disease are not overt as those in coalescent mastoiditis. The intact ear drum does not reflect the severity of bone-eroding disease within the mastoid. Since this disease is characterized not by exudative but by proliferative changes, there is no pus formation. The incidence of complications is high. Plain x-ray and CT scan do not specifically define the disease process, but bone scan indicates the bone invading nature of the mastoid infection. The osteoblastic reaction secondary to osteitis is demonstrated by high uptake of the isotope in the involved mastoid. Antibiotics may cure the disease, but in most cases surgery is unavoidable. PMID- 8225084 TI - [Recurrent intermittent priapism following priapism]. AB - Recurrent intermittent priapism is a rare sequela of prior priapism. We describe a patient with recurrent intermittent priapism 2 years after a prior event, without other causes of secondary priapism. We believe that this syndrome may be secondary to the initial ischemic event, resulting in alteration in penile adrenergic tone or the penile sinusoidal endothelium. PMID- 8225085 TI - [Secondary achalasia]. AB - Achalasia is usually a primary disorder of esophageal motility, but has been described in association with other pathological processes, such as malignancy. A 79-year-old man with achalasia secondary to gastric adenocarcinoma is presented. The differential diagnosis of secondary achalasia includes infectious and infiltrative disease and neuropathy, but mainly malignant diseases. The clinical criteria found for achalasia secondary to malignancy included older age at diagnosis, brief duration of symptoms, and weight loss. While upper gastrointestinal x-rays and computerized tomographic scanning may be helpful, the most reliable diagnostic tool is esophago-gastro-duodenoscopy. This is a terminal disease with short life expectancy. Yet making the correct diagnosis can save the patient from futile treatment with muscle relaxants and endoscopic balloon dilatation, the accepted therapeutic measures in primary achalasia. PMID- 8225086 TI - [Role of calcium antagonists in the treatment of atherosclerosis]. PMID- 8225087 TI - [Convulsive status epilepticus in children]. PMID- 8225088 TI - [Comparison of ovulation induction protocols for in vitro fertilization]. AB - In a retrospective study we evaluated 4 controlled ovarian hyperstimulation (COH) protocols for in vitro fertilization (IVF) treatment. Treatment cycles 4 through 8 were analyzed in patients, who in 3 previous attempts reached the stage of embryo transfer (ET) but did not conceive. The 426 recorded cycles included all indications for IVF, but not the male factor. The age range was 25-46 years. Patients underwent from 4 cycles (169 women) to 8 cycles (27 women). The COH protocols used included established combinations of menotropins, clomiphene citrate/hMG, and short and long protocols of GnRH analogs followed by hMG. There were no statistically significant differences in patient age, etiology and duration of infertility, or distribution of the various COH protocols among the groups of patients. No single protocol was significantly superior to any other when the pregnancy rate per cycle was assessed. The results did not justify changing a COH protocol which had lead to ET but not to pregnancy, for another protocol. PMID- 8225089 TI - [Evaluation of a new intravenous device]. AB - Problems associated with intravenous (IV) admixture preparation include contamination by microorganisms, human error and expense. To solve some of these problems, the Vialink system was developed by Travenol, Israel. This device enables enhanced sterility in clinical settings, since in the preparation of the admixture there is only a single penetration of the rubber seal of the vial. The dilution is performed immediately prior to administration, thereby reducing the possibility of wastage due to unused admixture. In addition, Vialink offers a means of double-checking the contents of the admixtures, since the emptied vial remains attached to the infusion bag. Results of our evaluation of this new system indicate that it offers substantial savings in preparation time and expense. It was suitable for 58.2% of all IV admixtures routinely hospital prepared. The Vialink system may be a useful strategy, as it improves the quality of the service provided by the health service team and hospital pharmacists to their patients, and results in savings in budget and manpower. PMID- 8225090 TI - [Fiber-enriched cereal for constipation and for hypercholesterolemia]. AB - A mixed-grain, high-fiber cereal (Disivit) prepared from oats, corn, wheat and soybean was used to treat 20 patients with chronic constipation and 22 with hypercholesterolemia in double-blind, cross-over trials. Disivit (50 g/d, containing 12.5 g dietary fiber) was given to the constipated patients for 2 weeks and then a low-fiber placebo for another 2 weeks, and similarly for the hypercholesterolemic patients. In those with constipation, the frequency of bowel movements increased significantly, stools became softer and laxative intake decreased. In hypercholesterolemic patients serum cholesterol decreased significantly, but only by 15%. Thus the fiber cereal appears to be a suitable treatment for constipation, while for hypercholesterolemia a larger dose or a longer period of treatment may be required. PMID- 8225091 TI - [Erythrocyte mean corpuscular volume of psychiatric patients]. AB - It has been observed clinically that the mean corpuscular volume (MCV) of erythrocytes is increased in psychiatric patients, but this phenomenon has not been investigated. We therefore compared the MCV of 177 patients in our psychiatric day care ward and in our psychiatric outpatient clinic. The MCV of these psychiatric patients, and especially that of the outpatients, were significantly greater than that of normal controls. No association was found between MCV and sociodemographic or clinical variables. The sensitivity of a high MCV level as a diagnostic for folic acid deficiency was 42.9%, with a specificity of 80.6%. The sensitivity of a high MCV level as a diagnostic for vitamin B12 deficiency was 23.1%, with a specificity of 79.4%. PMID- 8225092 TI - [Chromium in the treatment of clinical diabetes mellitus]. AB - The trace element chromium (Cr, tri-valent) is described in textbooks as indispensable in carbohydrate metabolism, but there have been no indications as to how to administer it and in which cases it might be helpful. We gave 243 diabetic patients Cr (200 mcg/d) to study its effect on blood glucose balance. 105 were Type 1 (IDDM) and 138 Type 2 (NIDDM). Cr reduced insulin, sulfonylurea or metformin requirements in 115 patients. The success rate was greater in those with NIDDM (57.2%) than in those with IDDM (33.6%). More women, of either type, reacted than men (62.5 vs 50% in NIDDM and 37.6 vs 28.6% in IDDM). A placebo was ineffective. We have not found any clinical signs indicating which patients might benefit from Cr. However, the IV insulin-responsiveness test might help detect patients who might benefit from addition of Cr to their food. Patients given the Cr supplement were told to reduce their usual dose of hypoglycemic agent to avoid hypoglycemia, and to increase the dose should their glycemia increase. PMID- 8225093 TI - [Traumatic asphyxia]. AB - Traumatic asphyxia is a commonly used designation of a syndrome related to severe compressive trauma to the thorax. It is characterized by cranial cyanosis, subconjunctival hemorrhage, vascular engorgement of the head, mucosal petechiae, hemoptysis, esophageal and rectal hemorrhage, hematuria, and varying degrees of cerebral dysfunction. By 1985 approximately 210 cases had been reported world wide. We present an illustrative case in a 24-year-old man. PMID- 8225094 TI - [Antimitochondrial antibodies and primary biliary cirrhosis]. PMID- 8225095 TI - [Hypertension in pregnancy--therapeutic approach]. PMID- 8225096 TI - [Postprandial lipoprotein metabolism and atherosclerosis]. PMID- 8225097 TI - [Diagnosis of aorto-enteric fistula]. PMID- 8225098 TI - [Early and late outcomes of cerebrovascular accidents and their rehabilitation]. PMID- 8225099 TI - [Geriatric assessment in acute care hospital]. PMID- 8225100 TI - [Importance of nocturnal penile tumescence (NPT) monitoring in the evaluation of erectile disorders]. PMID- 8225101 TI - [Heart failure due to diastolic dysfunction]. PMID- 8225102 TI - [Pet birds as a risk factor for lung cancer]. PMID- 8225103 TI - [Impact of Shahal (Cardiac Emergency Services) on daily life of subscribers]. AB - SHAHAL (Cardiac Emergency Services) provides professional 24-hour mobile emergency medical service monitored by intensive care nurses via a central computerized operations unit. All subscribers carry a "cardiobeeper" for transtelephonic transmission of a 3-lead electrocardiogram, and an automatic intramuscular self-injector containing 300 mg lidocaine. In this retrospective study we assessed the impact of SHAHAL on subscribers' mental stress, self confidence and ultimately, quality of life. Answers to a written questionnaire (n = 1034) and a personal interview (n = 55) were analyzed. It was shown that SHAHAL is effective in decreasing mental stress and improving self-confidence in the majority of subscribers. It was especially beneficial in women, the elderly, those of lower socioeconomic status, pessimists and chronic worriers, and the ill more than the healthy. Those whose functional capacities were very limited benefited less, but for them SHAHAL's function as a life-saving and rescue system is more valuable. Lastly, subscription to SHAHAL also improved the self confidence of the subscriber's family. PMID- 8225104 TI - [Idiopathic hypereosinophilic syndrome]. AB - The idiopathic hypereosinophilic syndrome is a heterogeneous group of disorders characterized by persistent eosinophilia of undetected cause, and multiple organ system involvement. The systems affected include the central and peripheral nervous, cardiovascular, respiratory and gastrointestinal systems and the kidneys, skin, muscles and joints. Treatment is mainly by immunosuppressive drugs to prevent organ system complications. Prognosis is variable, depending mainly on heart involvement. PMID- 8225105 TI - [Endoscopic drainage of pancreatic pseudocyst: endoscopic cysto-enterostomy]. AB - Pancreatic pseudocysts occur in 20% of cases of chronic pancreatitis. Spontaneous resolution is seen in about 9% of the cases, but most cysts persist and frequently cause complications, such as compression of adjacent organs or infection. Endoscopic cysto-enterostomy has recently been reported as a new nonsurgical invasive approach. We present a 40-year-old alcoholic man with a large pancreatic pseudocyst compressing the distal choledochus and pancreatic duct, who was treated successfully by endoscopic cysto-duodenostomy. PMID- 8225106 TI - [Crohn's disease of small bowel manifested by obstructive carcinoma]. AB - In ulcerative colitis the incidence of adenocarcinoma of the small bowel is higher than in the general population. More than 100 cases of carcinoma of the small bowel have been reported in patients with Crohn's disease of the same segment. However, obstructive carcinoma of the small bowel as the first manifestation of Crohn's disease has seldom been reported. We present such a case in which the first manifestation of otherwise silent Crohn's disease was intestinal obstruction by adenocarcinoma of the same segment. PMID- 8225107 TI - [Giant lipomas in unusual locations]. AB - 3 patients with giant lipomas located in unusual anatomic locations are described. All 3 cases had clinical features similar to those of the malignant counterpart, liposarcoma. All were operated on and the tumors removed. Follow-up ranged from 12 years, with no recurrence during that period. The treatment of choice for these lesions is surgical removal and long-term follow-up. PMID- 8225108 TI - [Diagnosis of Arnold-Chiari malformation by electronystagmography]. AB - 3 patients with Arnold-Chiari malformation with vertigo as the sole clinical presentation are described. 2 of them had been undiagnosed for many years despite repeated examinations, including CT scans. The third patient was diagnosed 2 weeks following the initial vertiginous attack. Downbeating nystagmus demonstrated by electronystagmography, indicating a pathological lesion at the craniocervical junction, was the key finding that led to the diagnoses. Following this finding, CT scan directed to the craniocervical junction showed the typical image of Arnold-Chiari malformation. The cases presented emphasize the important contribution of electronystagmography in the evaluation of vertigo due to intracranial lesions. However, computerized imaging examination remains the main diagnostic tool for intracranial pathologic processes. PMID- 8225109 TI - [Ectopic pregnancy: diagnosis and treatment]. PMID- 8225110 TI - [Genetics of inflammatory bowel diseases]. PMID- 8225111 TI - [Should serum cholesterol be checked in children?]. PMID- 8225112 TI - [The role of corticosteroids in chronic obstructive pulmonary disease]. PMID- 8225113 TI - [Diabetic retinopathy and pregnancy--interdependent risk factors]. PMID- 8225115 TI - [Euthanasia]. PMID- 8225114 TI - [Imaging blunt renal trauma]. PMID- 8225116 TI - [Janusz Korczak--physician (Part I)]. PMID- 8225117 TI - [The global fight against AIDS--are we advancing or retreating?]. PMID- 8225118 TI - The lacrimal fossa of cercopithecoidea, with special reference to cladistic analysis of Old World monkey relationships. AB - Re-examination of lacrimal fossa patterns in extant cercopithecoids indicates that the last common ancestor of Cercopithecini and Papionini, and hence of Cercopithecinae, probably retained a maxillary contribution to the lacrimal fossa, as did the common ancestor of Colobinae. Consequently, the presence of a maxilla-lacrimal fossa cannot be used to assess the subfamily affinity of Old World monkeys. In addition to being correlated with general facial lengthening, the derived, exclusively lacrimal pattern of Erythrocebus, Mandrillus, Papio, Theropithecus and some (but not all) guenons, macaques and mangabeys may be associated with extreme narrowing of the interorbital septum. Moreover, the derived condition may have evolved in response to independent exploitation of open country habitats as it enhances protection of the lacrimal sac and serves to reduce eye infection in terrestrial species. PMID- 8225119 TI - Behaviour and reproduction of the slender loris (Loris tardigradus) in captivity. AB - Observations were made on the behaviour of a group of slender lorises (Loris tardigardus) maintained in the Adelaide Zoo. Descriptions of their individual behaviour, social interactions, reproductive biology and development of young are given. The slender loris displays a more complex social life-style than has previously been reported, with huddling, marking activities and the behavioural complex of stretching/wrestling/allogrooming occurring daily. Evidence of the formation of a female coalition is described for the first time for this species. The low reproductive success of the group may have been attributable to the mixing of two Sri Lanka subspecies. Matings between L. tardigradus grandis and L. tardigradus tardigradus produced one sibling that reached maturity but did not breed. PMID- 8225120 TI - Middle Bartonian locality with Anchomomys (adapidae, primates) in the Spanish pyrenees: preliminary report. PMID- 8225121 TI - Prenatal testosterone effect on dermatoglyphic development in rhesus macaques. PMID- 8225122 TI - Feeding postures in the black howler monkey, Alouatta caraya. PMID- 8225123 TI - Home range of the brown howler monkey, Alouatta fusca, in a forest fragment of southeastern Brazil. PMID- 8225124 TI - A key for identifying individual female baboons under poor visibility conditions. PMID- 8225125 TI - Eye contact as a trigger of male sexual arousal in stump-tailed macaques (Macaca arctoides). PMID- 8225126 TI - Effects of dietary iron and folate supplementation on the physiological changes produced in weanling rats by sodium saccharin exposure. AB - Exposure of rats to high dietary levels of sodium saccharin (NaSac) started in utero produce physiological effects at 30 days post-birth that are similar to those found in pups of iron-deficient dams. These similarities suggest that some of the changes due to NaSac are secondary to iron deficiency. The present experiment investigated whether the effects of 7.5% dietary NaSac in the newborn rat could be prevented by dietary iron and/or folate supplementation. The NaSac related effects prevented by iron supplementation included anaemia, decreased serum iron and folate, increased serum cholesterol and triglyceride and increased serum vitamin E. Folate supplementation prevented NaSac-induced depression of serum folate and increase in serum vitamin E. Although bladder hyperplasia was increased by dietary iron and/or folate supplementation, the majority of the urinary chemistry changes associated with NaSac treatment were not affected. The results show that some physiological changes associated with NaSac treatment in the newborn rat may occur as a consequence of iron deficiency rather than a direct effect of NaSac treatment. These changes may be independent of the urinary and bladder effects, which are not reversed by iron supplementation. PMID- 8225127 TI - Renal clearance of domoic acid in the rat. AB - The renal clearance (Clr) of the seafood toxin domoic acid (DA) was investigated in the rat. Following cannulation of the right femoral artery, the left femoral vein and the bladder of anaesthetized rats, a single bolus injection of either [3H]DA, [14C]p-aminohippuric acid (PAH) or [3H]inulin was administered through the venous cannula. Blood samples were taken from the arterial cannula at 1, 2, 10, 30, 50, 70, 90, 110 and 130 min following injection, and urine samples were collected at 20-min intervals starting from the time of bolus injection. Based on plasma concentration-time profiles, the total clearances (Clt) for DA, PAH and inulin were 9.12, 33.17 and 7.50 ml/min/kg body weight, respectively. The Clr calculated from urinary excretion rates were not significantly different from the Clt. Probenecid significantly reduced the Clr of PAH but did not affect that of DA. When DA was given at doses of 0.5 ng, 0.5 mg and 2.0 mg/kg body weight, the pharmacokinetic parameters Clt, Clr, elimination-rate constant and apparent volume of distribution at steady state were not statistically different between doses. The entire dose of 3H was recovered in the urine by 160 min after dosing, and analysis of urine samples by HPLC confirmed that the radiolabel (3H) was associated predominantly with the parent form of DA. The results of the present study demonstrate that DA is cleared from plasma primarily through the kidneys. DA clearance occurs primarily by renal glomerular filtration since its Clt is comparable with that of inulin, is less than that of PAH and is not affected by probenecid. PMID- 8225128 TI - Bioassay of quinoline, 5-fluoroquinoline, carbazole, 9-methylcarbazole and 9 ethylcarbazole in newborn mice. AB - Quinoline and carbazole are among the more prevalent aza-arenes present as components of environmental pollutants. Both of these aza-arenes are hepatocarcinogenic to mice when administered in the diet. The hepatocarcinogenic potential of quinoline is consistent with its mutagenic activity in Salmonella typhimurium TA100 and potential to induce unscheduled DNA synthesis (UDS) in rat hepatocytes. Structure-activity studies with fluorinated quinolines indicate that the presence of a fluorine atom at the 5-position of quinoline may inhibit detoxification and result in enhanced genotoxic potency. Quinoline and 5 fluoroquinoline were assayed in newborn CD-1 mice at a total dose of 1.75 mumol to establish their relative tumorigenic activity. Liver tumours developed in 60 and 90% of the male newborn mice treated with quinoline and 5-fluoroquinoline, respectively. The majority of liver tumours observed among the quinoline-treated mice were classified as adenomas. In contrast, liver carcinomas developed in most of the male mice treated with 5-fluoroquinoline. Unlike the well established genotoxic potential of quinoline, there is limited evidence for carbazole having either genotoxic or carcinogenic activity. Whereas carbazole is not mutagenic towards several strains of S. typhimurium, both 9-methylcarbazole and 9 ethylcarbazole are active as mutagens in S. typhimurium TA100. Carbazole, 9 methylcarbazole and 9-ethylcarbazole were assayed in primary rat hepatocytes to assess their relative potential to induce UDS in rat hepatocytes; only 9 ethylcarbazole did so. These carbazole derivatives were also assayed in newborn CD-1 mice at a total dose of 1.75 mumol. Neither carbazole nor either of these 9 alkylcarbazoles produced a significant tumorigenic response in this bioassay system. PMID- 8225129 TI - Screening of heat-processed Finnish foods for the mutagens 2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline, 2-amino-3,4,8-trimethylimidazo[4,5 f]quinoxaline and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine. AB - The concentrations of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and DiMeIQx (i.e. the sum of 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline and 2-amino-3,7,8 trimethylimidazo[4,5-f]quinoxaline) were determined as their 3,5 bis(trifluoromethyl)benzyl bromide derivatives in commercial heat-processed daily Finnish foods using the gas chromatography-mass spectrometry technique with negative chemical ionization. With this technique it was possible to detect picogram quantities of halogenated aminoimidazoazaarene (AIA) derivatives. The mutagenic activities of the food samples were tested using the Ames Salmonella assay. The mutagenicities and amounts of the AIA compounds analysed varied considerably in the food samples. All the flame-broiled fish samples and the majority of the grilled pork and chicken samples were positive in the test; of all the food samples tested fish samples had the highest mutagenic activity. Industrially produced meat patties were non-mutagenic. The amounts of MeIQx, DiMeIQx and PhIP in grilled products that exhibited mutagenic activity varied in the range 0.04 to 0.4, 0.03 to 0.2 and 0.5 to 3.8 ng per g cooked food, respectively. Some of the highly mutagenic flame-broiled fish samples contained only PhIP, the amount of which varied from 0.5 to 5.5 ng per g fish. One of the samples also contained a small amount of MeIQx. None of the non-mutagenic meat patties contained detectable amounts of MeIQx, DiMeIQx or PhIP. In this screening study evaluation of the occurrence of AIA compounds was carried out in a relatively wide range of commercial heat-processed Finnish foods. Considerable differences between equivalent products from different manufacturers were found in many cases. This variation indicates that industrial processing of food has a marked effect on the mutagenic activity of the product. PMID- 8225130 TI - Effects of 1-isothiocyanato-3-(methylsulfinyl)-propane on xenobiotic metabolizing enzymes in rats. AB - The glucosinolate hydrolysis product 1-isothiocyanato-3-(methylsulfinyl)-propane (IMSP), also known as iberin, is consumed in the average human (US) diet at approximately 1 mumol/kg/day. The chemoprotective effects observed with the consumption of cruciferous vegetables may be due to the presence of specific glucosinolate hydrolysis products either within the crucifers, or formed after ingestion of the crucifers. The mechanism of chemoprotection may be through selective induction of components of Phase II xenobiotic metabolizing enzymes. The influence of repeated administration of low concentrations of IMSP by gavage on components of Phase I and Phase II xenobiotic metabolizing systems was examined in the liver and small intestine of male Fischer 344 rats. Doses of 1, 10 and 100 mumol IMSP/kg, administered by gavage for 7 days, did not alter weight gain, or hepatic and renal weights, relative to body weight, and did not cause any histological lesions. Intestinal glutathione S-transferase (GST) activity and NAD(P)H:quinone reductase (QR) activities were significantly elevated to 3.1 and 8.1 times control values, respectively, at the 100 mumol/kg dose only. The administration of IMSP at 1, 10 or 100 mumol/kg had no significant effect on hepatic Phase I enzymes activities (cytochrome P-450 concentrations, ethoxycoumarin O-deethylase [ECD] and aminopyrine N-demethylase [AND] activities) or Phase II enzyme activities (GST, QR and UDP-glucuronosyltransferase [UDP-GT] activities towards 1-naphthol or 4-hydroxybiphenyl), at any of the doses tested and no effect on intestinal enzyme activities at doses below 100 mumol IMSP/kg. It is concluded that IMSP does not have a significant influence on induction of the Phase I or Phase II xenobiotic metabolizing enzymes in rats when tested at doses approximating those found in the human diet. PMID- 8225131 TI - Antimutagenic effects of casein and its digestion products. AB - The antimutagenic potential of casein was investigated using several mutagens, including some food-related mutagens, and the Salmonella/microsome and Escherichia coli DNA-repair tests. The effect depended on the type of mutagen and the incubation time of casein with the mutagens. Casein was very effective against benzo[a]pyrene, N-methylnitrosourea and nitrosated 4-chloroindole, and was less effective towards sodium azide and N-nitroquinoline-1-oxide (NQO). Preincubation increased the antimutagenic potential of casein towards NQO. Heating of casein (up to 20 min at 130 degrees C) did not alter its antimutagenic capacity. The effect of pepsin hydrolysis under simulated gastric conditions on the antimutagenic capacity of casein was tested with sodium azide and NQO in the Salmonella/microsome test. The peptides formed were separated by ultrafiltration or by isoelectric precipitation of casein, and were characterized by HPLC size exclusion and Kjeldahl analysis. The antimutagenic potential of casein increased with pepsin hydrolysis; this increase was due to the peptides formed and might be explained by a better accessibility of casein peptides for interaction with mutagens. The antimutagenic potential of pepsin-hydrolysed casein towards sodium azide was observed over the whole dose-response curve. PMID- 8225132 TI - In vitro toxicity evaluation of a product obtained from carmoisine using Tetrahymena pyriformis cells. AB - The toxicity of carmoisine and of its metabolites after interaction with media components, was studied using the single cell system of Tetrahymena pyriformis. Carmoisine was not toxic to T. pyriformis. However, when autoclaved with media components such as bactopeptone or yeast extract, it gave rise to product(s) that completely inhibited the growth of T. pyriformis. Addition of glucose to the bactopeptone-carmoisine mixture before autoclaving significantly enhanced the formation of the toxic product. A similar effect was observed with other reducing sugars and with compounds containing sulfhydryl groups. The results indicate that carmoisine may undergo initial reduction, followed by a series of reactions leading to the formation of the toxic component(s). PMID- 8225133 TI - Effects of riboflavin deficiency and riboflavin administration on carcinogen-DNA binding. AB - A study was conducted to assess the effects of riboflavin deficiency and riboflavin supplementation on carcinogen-DNA binding. After 12 wk on a riboflavin sufficient or a riboflavin-deficient diet male Wistar rats were administered 3H labelled benzo[a]pyrene (BP) ip. [3H]BP was given either at a uniform dose of 450 muCi/rat irrespective of body weight or at a dose adjusted to body weight. After 17 hr the animals were killed, various organs were dissected and the level of [3H]BP bound to DNA was quantified in organs that are known to be the seats of drug metabolism (i.e. the liver, lungs and intestinal mucosa). In a separate experiment, the effect of riboflavin supplementation on BP-DNA binding was also investigated. When [3H]BP was administered at 450 microCi/rat, BP-DNA binding was markedly increased in the livers and intestinal mucosae of the pair-fed and deficient groups compared with controls. With the administration of [3H]BP adjusted to body weight, no differences in BP-DNA binding between groups were observed in any tissue. However, on administration of riboflavin there was a decrease in the level of [3H]BP bound to DNA in almost all tissues, especially in the lungs, where the reduction was significant. The results suggest that undernutrition/riboflavin deficiency may increase the risk of carcinogenesis by way of an increase in carcinogen binding, which however can be reversed by riboflavin supplementation. PMID- 8225134 TI - Determination of combined benzidine in FD & C yellow no. 5 (tartrazine), using a highly sensitive analytical method. AB - 53 samples of FD & C Yellow No. 5 (tartrazine; Colour Index No. 19140) were examined for combined benzidine. These samples, which represent separate lots from 12 dye distributors, were submitted to the US FDA for certification between 28 February 1990 and 27 June 1991. A method was developed to reduce the dye matrix with dithionite so that combined benzidine present in the form of azo or disazo dyes would be converted to free benzidine. Reduction was followed by extraction, diazotization and coupling with pyrazolone T, and the total benzidine present was quantitated as benzidine-pyrazolone T disazo dye (BZPT) by HPLC with detection at 500 nm. The limit of quantitation for benzidine in FD & C Yellow No. 5 by this method is 5 ng/g. 25 samples of FD & C Yellow No. 5 were found to contain 7-83 ng/g of combined benzidine that was released by dithionite reduction. 23 of these samples were from the same manufacturer. The identify of the BZPT from two FD & C Yellow No. 5 samples was confirmed by spectral analysis using HPLC with a photodiode array detector. PMID- 8225135 TI - Challenge assay in vitro using lymphocyte blastogenesis for the contact hypersensitivity assay. AB - To confirm positivity in routine guinea pig studies, contact allergenicity was investigated by a challenge assay in vitro using a co-culture of autologous lymphocytes passed through a nylon-wool column and antigen-presenting cells (APCs) modified with or without antigen. Proliferation of the lymphocytes primed with ovalbumin and/or 2,4-dinitrochlorobenzene was antigen specific and dependent on the presence of APCs (peripheral blood monocytes, splenic macrophages and macrophages induced by liquid paraffin). For another nine haptens, primed lymphocytes proliferated significantly more than control lymphocytes; the stimulation index (SI; ratio between [3H]methylthymidine ([3H]TdR) incorporation of lymphocytes with antigen-modified APCs and [3H]TdR incorporation of lymphocytes with APCs not modified by antigen) was 1.6-4.8 in sensitized animals whereas it was about 1.0 in control animals. Sodium dodecyl sulfate did not cause lymphocyte proliferation. The SI value in vitro was correlated with both the positive rate in vivo (r = 0.736) and the mean response score in vivo (r = 0.645). Thus, it was possible to confirm that positivity in routine experiments was a true sign of allergy. A combination of this assay and short-term animal studies would provide an efficient assessment of the allergic potential of chemicals. PMID- 8225136 TI - Bioassay of mycotoxins using terrestrial and aquatic, animal and plant species. AB - The role of bioassay in the diagnosis of mycotoxicoses in farm animals is discussed. Methods for detecting mycotoxin contamination of animal feeds using biological species (aquatic and terrestrial animals and plants but excluding bacteria and yeast) are reviewed. Factors that need to be considered when choosing and using particular bioassay procedures in a feed-screening programme are discussed in relation to the occurrence of false positives and false negatives. PMID- 8225137 TI - Postmortem serum selenium concentrations and their possible etiological role in sudden infant death (SID). AB - The importance of selenium (Se) deficiency in the pathogenesis of human diseases such as Keshan Disease has been extensively studied. It is possible that low Se levels could cause immunosuppression and be an etiological factor in Sudden Infant Death (SID). We investigated 50 serum samples (40 SID and 10 non-SID victims) by atomic absorption spectrometry. The results show that there is no evidence of a serum selenium deficiency in SID-victims in the region of Aachen. A relationship between selenium concentration and the infant immune system still remains speculative and 21 samples even showed increased serum levels. It could be necessary to define a 'local' normal range by examining a greater number of healthy infants. PMID- 8225138 TI - Estimation of stature from footstep length. AB - Estimation of stature from various foot and shoe measurements with the help of statistical methods is well known. However, very little work has been reported on stature determination from foot and shoe stride lengths. As the stride length of a person is related to the height of the person and speed at which he is walking, certain constants and equations for stature determination from stride length can be formulated. PMID- 8225139 TI - Estimation of local vital reactions in severely burned tissues of guinea pig skin using C3a or C3a desArg as a marker. AB - We observed changes in levels of anaphylatoxin C3a and/or its desArg (C3a/C3a desArg) peptides in the local site of severely burned skin tissues of guinea pigs using immuno-Western blotting methods. The C3a/C3a desArg peptides, which were probably generated during complement activation, were detected at significant levels from 30 min to 72 h following burn injury in an area limited to 3 mm from the wound edge. Levels of these peptides showed a tendency to be highest in that area 24 h after burn infliction. In postmortem injuries, C3a/C3a desArg peptides could not be detected. These peptides were detected in antemortem wounds up to at least 2 days at 22 degrees C and up to 3 days at 4 degrees C after death, although decreases in levels were found. Lower concentrations of these peptides were also found in postmortem burns in which postmortem hypostasis appeared strongly. These results suggest that, except for injuries of the area with obvious postmortem hypostasis, detection of C3a/C3a desArg can be useful for estimation of vital reactions in many kinds of wounds during inflammation. PMID- 8225140 TI - Immunoassay evidence for fentanyl in hair of surgery patients. AB - Head hair samples obtained from surgery patients who received fentanyl during anesthesia were analyzed by immunoassay for the presence of fentanyl. Thirteen hair samples were collected from patients following intravenous administration of 1-6 mg of fentanyl. Additional hair samples were collected following the administration of 0.18 and 0.38 mg of sufentanil to 2 patients. The elapsed time after drug administration for all patients ranged from 7 to 273 days. Twenty control hair samples also were collected from staff members who reported no surgery or anesthesia during the previous year. All samples were initially washed with methanol, followed by extraction with methanol and reconstitution in citrate buffer. Analysis of wash and extract fractions was performed by radioimmunoassay (Coat-A-Count Fentanyl assay). Segmental analysis was performed on the surgery patients' hair samples. Eight of the fentanyl patients' hair samples contained fentanyl concentrations (equivalents) of 0.13-0.48 ng/10 mg of hair in the 'root' end. Fentanyl concentrations in the 'tip' segment were lower than those found in the 'root' segment with the exception of 1 subject whose hair sample had been collected only 7 days after surgery. The remaining 5 patients had fentanyl concentrations similar to those determined for the control subjects hair samples (0-0.08 ng/10 mg, n = 19). No correlation between hair fentanyl concentration and administered dose was found for the 13 fentanyl subjects. Both sufentanil subjects' hair samples tested negative. One control subject who had experienced environmental exposure to fentanyl had a fentanyl concentration of 0.29 ng/10 mg in the extract and 0.63 ng/10 mg in the wash fraction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225141 TI - Purification of blood-group substances in human hepatic bile (HHB) and immunological characterization of anti-HHB serum. AB - Blood-group substances (BGS) in human hepatic bile (HHB) of blood type A were purified 3657-7314-fold by ethanol sedimentation and column chromatography with DEAE Sephadex A-50 and Concanavalin A Sepharose 4B. An anti-HHB serum was raised by immunizing rabbits with the purified BGS in the HHB. Cross-reactions of the native anti-HHB serum with such human materials as type A blood cell, plasma, seminal fluid and saliva were eliminated by stepwise sequential absorptions with type A red blood cells, type O seminal fluid and type AB plasma. Precipitin titers of the native anti-HHB serum were x 320 and x 80 and those of the absorbed anti-HHB serum x 80 and x 20 against the immunogen and HHB, respectively. Organ and species specificity of the absorbed anti-HHB serum was checked by counter immunoelectrophoresis. It reacted specifically only with the immunogen and HHBs of various ABO blood types. No reactions with animal gallbladder biles were observed. In practical examinations, the specific anti-HHB serum identified 15 out of 20 (75%) HHB samples and 41 out of 46 (89%) human faeces samples. No reactions with animal gallbladder biles and animal faeces were observed. PMID- 8225142 TI - [Medical consultation--the key factor in andrologic examination]. PMID- 8225143 TI - [Long-term storage of cryopreserved semen. Quality control of cryopreserved semen in three year intervals]. PMID- 8225144 TI - [Disorders of fertility in men: basic andrologic diagnosis. Which routine studies are essential in initial andrologic examination?]. PMID- 8225145 TI - [Weight loss with alternative diets. Principles, philosophy, side effects, reliability. Part 7: Alternative diets (M)]. PMID- 8225146 TI - [Conservative therapy of backache. Part 5: TENS, acupuncture, biofeedback, traction, cryotherapy, massage and ultrasound]. AB - In addition to the major therapeutic modalities for use in back pain already discussed, a number of other forms of treatment are also available and in common use some of which at least have a solid scientific basis. Recent data suggest that subthreshold TENS is no effective treatment for low back pain. Whether this is also true of higher-dose TENS is not yet clear. Electro-acupuncture, in contrast, does appear to be effective, but not biofeedback or traction. Other commonly employed treatment modalities, such as cryotherapy, heat, massage or ultrasound cannot definitively be assessed at present, since controlled trials have not been carried out. PMID- 8225147 TI - [Oral combination therapy of zoster neuralgia. Pain reduction by 1-adamantanamine sulfate and carbamazepine per os]. AB - In four patients hospitalized with severe neuralgic complaints in conjunction with a Zoster infection, the pain-relieving effect of oral 1-adamantanamine sulfate used in combination with carbamazepine was studied. From the results obtained, the oral administration of 1-adamantanamine sulfate also appears to have a reliable analgesic effect, so that ambulatory treatment is readily possible. PMID- 8225148 TI - [Weight reduction with alternative diets. Principles, philosophy, side effects and uses. Part 8: Alternative diets (N-Q)]. PMID- 8225149 TI - [Moxonidine vs. captopril in minor to intermediate hypertension. Double-blind study of effectiveness and tolerance]. AB - In a randomized double-blind study, the antihypertensive efficacy and tolerance of the imidazoline receptor agonist, moxonidine, were compared with those of the ACE inhibitor, captopril. Included in the trial were 50 ambulatory patients with mild-to-moderate hypertension, who were treated for 4 weeks with either 0.2-0.4 mg moxonidine, or 25-50 mg captopril daily. Both substances clearly reduced hypertension; no statistically significant difference was seen between the two groups. Under moxonidine, the mean blood pressure while seated decreased from 176/101 mm Hg to 155/91 mm Hg by the end of treatment; under captopril the corresponding figures were 170/99 and 150/89 mm Hg. Minor transient side effects, for the most part with a doubtful relationship to the treatment, were seen in 5 patients (20%) of the moxonidine group, and in 8 patients (32%) of the captopril group. Thus, in this study, moxonidine and captopril proved to have equivalent antihypertensive efficacy and good tolerability. PMID- 8225150 TI - [Olfactory hallucinations in depression]. AB - Olfactory hallucinations are rarely mentioned as a symptom of depressive disorder. This paper presents a review of the literature including a conceptual clarification and epidemiological aspects with an emphasis on the significance of olfactory hallucinations for psychopathology, in particular with respect to sensory perception and body self-awareness. The close relation to the olfactory reference syndrome is pointed at. Olfactory hallucinations are considered important for the biological interpretation of affective disorders. In this regard disturbed olfactory perception can be understood as an example of disturbed brain function in the state of depression. Olfactory symptoms are shown to be elements connecting primary affective disorders with other neuropsychiatric diseases as well as with neuroanatomical and neurophysiological models. A primary role is accordingly attributed to the temporal lobe epilepsies in which olfactory hallucinations as well as affective disturbances occur. The need for further investigation in this field is pointed out. PMID- 8225151 TI - [Acute prognosis and differentiation of various cerebral infarct groups in comparison with cerebral hemorrhage]. AB - For clinical trials classification of stroke should be possible at the bedside by simple methods that are available every where. In this study are 1105 patients with every first ischaemic strokes and 130 patients with intracerebral haemorrhages. The differences between severity of clinical symptoms, outcome and risk factors of intracerebral haemorrhages, ischaemic stroke caused by cerebral microangiopathy, ischaemic stroke combined with extracranial carotid stenosis, cardiogenic brain embolism and atherothrombotic stroke, were analysed. Intracerebral haemorrhages show the poorest outcome of all groups (mortality 23.8%), due to increased intracranial pressure. Cardiogenic brain embolism is more frequent in older women (mean age 77.8 y.). Main risk factor is atrial fibrillation with absolute arrhythmia. The outcome of this group is the worst of all subgroups of ischaemic stroke and survivors most often in need of institutionalization. Patients with ischaemic stroke combined with extracranial carotid stenosis are significantly younger (mean age 67.6 y.), predominantly male, and smokers. Their mortality is low (0.63%), but recovery of paresis is slower than in other subgroups. Ischaemic strokes caused by cerebral microangiopathy with hypertension as main risk factor recover most quickly but acute mortality is higher than in ischaemic stroke combined with extracranial carotid stenosis because of higher age (mean age 74.5 y.). Institutionalization is more frequent too because of higher incidence of dementia in this subgroup. The main prognostic factors of all groups are age and severity of clinical symptoms. A special subgroup are infratentorial ischaemic strokes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225152 TI - [Life threatening situations caused by bolus aspiration in psychiatric inpatients -clinical aspects, risk factors, prevention, therapy]. AB - Five cases of life-threatening bolus aspiration are described. Four patients could be rescued; an 48 year old woman died of an reflectoric bolus death, also called "cafe coronary". A review of the literature and the special conditions of our examples allow to define different predisposing factors for bolus aspiration: severe psychiatric illness, abnormal eating behaviour, local or spread brain lesions, dysphagia, old age, multimorbidity and newly institutionalised persons. Possible risk exacerbations by psychotropic drugs are discussed. The special threat to psychiatric patients is shown. Prevention of these dangerous situations consists of an individual dysphagia risk screening and of following certain rules concerning eating and medication. All staff members in psychiatric hospitals should be trained in life-saving techniques in bolus incidents. PMID- 8225153 TI - [Clozapine in long-term treatment of schizophrenic patients]. AB - Clozapine has advantages over standard antipsychotics in refractory schizophrenia. Studies on the efficacy of clozapine in the maintenance treatment are sparse and suffer from methodological limitations. Despite this fact clozapine ranked second in a survey dealing with the preference of this antipsychotic in the long-term treatment of schizophrenia. Doctors report on using a mean of 130 mg/d in this indication which is considerably less than the doses used in most of the published long-term trials. The discrepancy between the popularity of clozapine and the lack of sound empirical data on its long-term efficacy is discussed. PMID- 8225154 TI - [Follow-up study on post-transfusion hepatitis in the Kyushu University Hospital]. PMID- 8225155 TI - The acute toxicity of allyl chloride by subcutaneous injection in mice. AB - Male ICR mice were administered allyl chloride at dose of 496 mg/kg, 600 mg/kg, 720 mg/kg, 864 mg/kg or 1037 mg/kg by a single subcutaneous injection. Sixteen of 25 mice died by the 7th day after the injection and LD50 was calculated 621 mg/kg body weight (95% C.I.: 522-739 mg/kg). A marked congestion with severe hemorrhage and edema were observed in the lung. Liver and kidney damages were also found, and these were characterized by the dilated sinusoids, degenerative change of hepatic cells, and focal necrosis in the liver; and necrosis of epithelium in convoluted tubules of the kidneys. Nine mice have survived by the 7th day after the injection and all of them showed a various degree of damages in the testes. The testicular lesions could be classified into two types. One type of the lesion was characterized by degeneration and exfoliation of germ cells, appearance of polynuclear giant cells in the seminiferous tubules, and mild proliferation of Leydig cells in the interstitium. In another type of the lesion, all type of cells in tubules, including Sertoli cells, and Leydig cells became necrotic. This is the first study reporting the testicular toxicity of allyl chloride. PMID- 8225156 TI - [The changes in Ca(2+)-sensitivity of rabbit skeletal myofibrillar ATPase activity under various conditions]. AB - Effect of the composition of the reaction mixture on the Ca(2+)-sensitivity of myofibrillar ATPase of rabbit skeletal muscle was investigated. The Ca(2+) concentrations necessary for the half maximum ATPase activation were 2.95 x 10( 6) M at 50 mM KCl, 4.37 x 10(-6) M at 90 mM KCl and 5.25 x 10(-6) M at 130 mM KCl. Thus the Ca(2+)-sensitivity was definitely lower at higher KCl concentrations. The Ca(2+)-sensitivity was about 2 times higher at pH 7.3 (pCa50 5.65) compared to the Ca(2+)-sensitivity at pH 6.8 (pCa50 5.36). The change in the Mg-ATP concentration between 0.5 mM and 3.5 mM, did not significantly affect the Ca(2+)-sensitivity of myofibrillar ATPase, but affected biphasically the maximum ATPase activity. These change in the Ca(2+)-sensitivity at various conditions is considered to be due mainly to the property of troponin C, the Ca(2+)-receptive protein of myofibrils. PMID- 8225157 TI - Acyclovir-resistant herpes zoster encephalitis successfully treated with vidarabine: a case report. AB - A 78-year-old man developed herpes zoster virus (HZV) encephalitis. Initially, treatment with aciclovir (750 mg per day) improved CSF cell count and protein level. During the treatment, however, encephalitis in the patient deteriorated in spite of the treatment with aciclovir, suggesting that HZV in the patient had become resistant to aciclovir. Subsequent treatment with vidarabine (600 mg per day, for 15 days) resulted in dramatic improvement in CSF pleocytosis. About two months after the discontinuation of vidarabine, the CSF cell count was normal. The patient became alert gradually, but his amnestic syndrome remained unchanged. Vidarabine may be recommended in the treatment of HZV encephalitis when aciclovir is not effective. PMID- 8225158 TI - [Alcoholic liver injury accompanied with various metabolic and endocrinological disorders--a case report]. AB - We report a young female case of alcoholic liver injury accompanied with various metabolic and endocrinological disorders. A 29 year-old woman was admitted because of general fatigue and hyperlipidemia. She was a heavy drinker. Laboratory data on admission revealed liver dysfunction and hyperlipidemia (type II b) with a quite high serum gamma-glutamyltranspeptidase (gamma GTP) level. The microscopic finding of the liver biopsy specimen showed fatty metamorphosis and ballooning of hepatocytes, and she was diagnosed as heavy alcoholic liver injury. The endocrinological examination revealed the elevated plasma cortisol level, though the urinary 17-hydroxycorticoids (17-OHCS) and 17-ketosteroids (17-KS) excretion and the plasma adrenocorticotropic hormone (ACTH) level were reduced. Cortisol secretion showed the normal circadian rhythm and the normal response to ACTH provocation. The levels of plasma triiodothyronine (T3), thyroxine (T4), and thyroid stimulating hormone (TSH) were also reduced. These endocrinological and metabolic disorders were normalized in company with recovery of the liver function by temperance, diet therapy and nutritional education. Thus, these abnormalities were considered to be resulted from the alcoholic liver injury and the effect of the ethanol to the hypothalamic-pituitary system. PMID- 8225159 TI - [Current outcomes and future directions in the chemotherapy of lung cancer]. PMID- 8225160 TI - Deep breathing test (DBT) in predicting white coat hypertension. AB - We have evaluated the diagnostic value of deep breathing test (DBT) in detecting the patients with white coat hypertension on the outpatient basis. Thirty patients with untreated mild to moderate hypertension underwent 24-hour ambulatory blood pressure monitoring. Those who had a mean 24-hour blood pressure less than 135/80 mmHg were categorized as having white coat hypertension and the remainder were categorized as sustained hypertension. These two groups were compared with regard to the differences of office blood pressures before and after DBT (i.e., deep breathing 5 times for a minute in sitting position). Although the office systolic pressure significantly fell after DBT in both groups (p < 0.001 vs p < 0.05), there was no significant difference (p = 0.27) between the two groups. On the other hand, the office diastolic blood pressure significantly fell in the white coat hypertensives in contrast with no meaningful fall of it in the sustained hypertensives (p < 0.01 vs p = 0.66). At the cutoff level of -3%, -5% and -10% of the differences in office diastolic blood pressure before and after DBT, the sensitivity for the presence of white coat hypertension were 64.7%, 58.8% and 29.4%, respectively, and the specificity were 61.5%, 84.6% and 100%, respectively. These findings demonstrate that the deep breathing test is useful for identifying white coat hypertension in the outpatient clinic. PMID- 8225161 TI - Serum inhibition of antibiotic-dependent L-form growth in Staphylococcus aureus: likely involvement of serum high-density lipoprotein. AB - The L-form growth of Staphylococcus aureus induced by ampicillin or fosfomycin was found to be severely depressed by horse or bovine serum included in the culture medium. Fractionation of horse serum revealed that the inhibitory activity was copurified with high-density lipoprotein through floating ultracentrifugation, gel filtration, and isoelectric focusing. It was also shown that the inhibition was reversible: ampicillin-induced L-form cells lost their ability to form L-form colonies when mixed with the lipoprotein fraction or serum before plating, but the colony-forming ability was restored to the original level by treatment of the mixture with a protease, while simple dilution was ineffective. A mechanism for the growth inhibition remains to be clarified. PMID- 8225162 TI - A rapid radioassay and some properties for leukotriene C4 synthase. AB - Incubation of glutathione, containing 35S-glutathione (GSH), and leukotriene (LTA4) in the presence of LTC4 synthase produced 35S-LTC4. The LTC4 was ether extracted and detected by ultraviolet absorbance (280 nm) after reverse phase high performance liquid chromatography (RP-HPLC) separation. The 35S-LTC4 was quantitated by scintillation counting. LTC4 production increased in a dose dependent manner for GSH concentrations from 10(-8) to 10(-2) M. On the contrary, the 35S detected in the ether phase and the 35S-LTC4 recovered from RP-HPLC decreased inversely with the concentration of unlabeled GSH added but did not with the addition of L-cysteine. This decrease in 35S-LTC4 production versus increased GSH levels results from a direct competition between unlabeled GSH and 35S-GSH reacting with the LTA4. Incubation of GSH (0.5 mM), containing 35S-GSH (0.3 uCi, 10 nM) and LTA4 (10 microM) with LTC4 synthase produced 35S-LTC4 at a linear rate over 5 min. The 35S-LTC4 increased in proportion to the LTC4 synthase added. Therefore, we have demonstrated that LTC4 synthase activity can be monitored by a simple radioassay procedure that measures 35S counts in ether extracts from incubation mixtures. This reliable and rapid assay procedure promises to facilitate characterization and purification of LTC4 synthase, a membrane-bound GSH S-transferase of unique specificity. PMID- 8225163 TI - ["AIDS". Advance of treatment and care of human immunodeficiency virus (HIV) infected persons by medical doctors, dentists, nurses and paramedics without discrimination from other patients. Introduction]. PMID- 8225164 TI - [HIV vaccine trials]. AB - There has been significant progress on several candidate vaccines against HIV infection over the past few years, i.e., live recombinant virus vaccines, inactivated virus vaccines, and purified antigens (subunit vaccines). There have been many phase I studies in healthy volunteers uninfected with HIV, as well as phase II trials with high-risk uninfected volunteers. Most of them focused on productions of neutralizing antibodies and cytotoxic T lymphocytes (CTLs) against HIV Env proteins. In addition, host immune responses, particularly cell-mediated immunity, were shown to be a major immune defense mechanism which may provide resistance to HIV spread leading to AIDS, indicating the possibility to develop prophylactic vaccine which may prevent the onset of AIDS in HIV carriers. However, various studies have implicated the concurrent activities of several HIV proteins, such as Env gp120, Env gp41, Tat, and Nef, in the induction of immunodeficiency. Therefore, the role of HIV-1 proteins in inducing immunodeficiency after HIV vaccination should be considered. Here, I introduce a recombinant BCG (rBCG) vaccine which expresses the immunodominant gp120 epitope. The tuberculosis vaccine strain Mycobacterium bovis BCG is a widely used vaccines with a low rate of serious complications. To develop a vaccine expressing an immunodominant epitope in the gp120 V3-loop, we used our system to express and secrete an epitope which was fused with an extracellular alpha antigen of Mycobacterium kansasii. Balb/c mice inoculated with this rBCG, rapidly induced HIV-specific CTLs. Target cell lysis was restricted to the murine class I major histocompatibility complex, H-2d.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225165 TI - [AIDS and opportunistic virus infections]. AB - The opportunistic herpesvirus infections in AIDS are often life-threatening and the patients have lowered Epstein-Barr virus (EBV)-specific T cell cytotoxicity. On the other hand, the asymptomatic human immunodeficiency virus (HIV) carriers have potentially lowered EBV-specific cytotoxic T cell function, which is shown by exposure of the lymphocytes to an immunosuppressive substance. The EBV specific T cell cytotoxicity may therefore tell the timing to cope with such opportunistic viral infections. PMID- 8225166 TI - [AIDS-related malignancy]. AB - Although Kaposi's sarcoma (KS) has been considered a rare disease, the disease is well known at present since the onset of AIDS in 1981. The characteristics of AIDS-associated KS are a multifocal, widespread distribution that may involve lymph node, gastrointestinal tract, and visceral organs. KS may be the first sign of HIV-infection, but it can also arise in some patients who lack evidences of immune impairment. The more effective chemotherapy of AIDS-associated KS is low dose-ABV-combination (adriamycin, bleomycin and vincristine) and its response rate is about 80%-90%. The second cancer that occurred in the AIDS-related immune impairement is malignant lymphoma. Approximately 90% of AIDS-related malignant lymphoma reported have been of high grade, B-cell types, including B immunoblastic type and small non-cleaved cell lymphoma. They have another distinguishing feature that is wide spread extent of disease at presentation, with extranodal involvement recorded in 80% to 90% of all patients. The most common sites of involvement are CNS (central nervous system) (32%), gastrointestinal tract (26%), bone marrow (25%) and liver (12%). It was reported that the median CE4 count in patients with primary-CNS lymphoma was 37 cells/dl, versus 189 cells/dl in those with systemic disease. It is important to note that approximately 17% of leptomeningeal disease is asymptomatic. The recommended treatment of AIDS-associated lymphoma by Levine is a low-dose modification of the M-BACOD (bleomycin, doxorubicin, cyclophosphamide, vincristine, dexamethasone, cytosine arabinoside, azidothymidine and helmet field radiotherapy). A complete remission (CR) rate of 46% was achieved. The median survival time of CR patients was 15 months.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225167 TI - [Two cases with acquired immunodeficiency syndrome in our hospital]. AB - We report two cases of AIDS whom we have recently experienced. One patient was a 54-year-old man who admitted our hospital due to third degree burn. In this case, we did not know whether or not he was suffered from AIDS, when he was delivered by the ambulance. In autopsy, pneumocystis carinii pneumonia and renal tuberculosis were found in addition to marked decrease of T cells in lymph nodes. The other patient was a 40-year-old man with remarkable symptoms of central nervous system. Route of infection of HIV is unknown. He had dementia, left hemiplegia, bulbar palsy, progressed to rigid decorticate posture and died of respiratory arrest due to involvement of the brain stem, despite of treatment including use of 3'-azido-2',3'-dideoxythymidine (AZT). Magnetic resonance (MR) images showed progressive cerebral atrophy and a diffuse high signal intensity area of cerebral white matter on T2-weighted MR images, suggesting the diagnosis of HIV-induced encephalopathy. PMID- 8225168 TI - [Issues in the AIDS medicine based on the analysis of medical worker's recognition of the disease]. AB - We analysed problems on the front line of the AIDS medicine through our experience of a with hemophilia A, who had HIV infection developed to ARC and the AIDS and in addition, had non-Hodgkin's lymphoma in his rectum and died of pneumonitis caused by pneumocystis carinii. We have learned from the poll on medical worker's recognition of the disease through the experience of this case that the nurses who were concerned with the care of this patient did not have opinions against acceptance of patients with AIDS, although they have a fear, from the professional point of view, to a possibility of infection of the virus from patients with AIDS. What should be done in the practical part of the AIDS medicine are (1) a repetitive education of accurate knowledge of the disease, (2) completion of practical guide lines for medical care of patients with AIDS and (3) legal establishment of an insurance system for accidental infections of HIV to medical or paramedical staffs. PMID- 8225169 TI - [Preventive measures against HIV infection]. AB - It is reported that approximately 900 persons with human immunodeficiency virus (HIV) are present in Japan. Among them, almost 300 persons developed acquired immunodeficiency syndrome (AIDS). However, we have to assure that more persons infected with HIV or patients with AIDS, which are not registered, appear to exist even in Japan like in other countries. Accordingly, it is just a matter of time for us to treat them in any hospital. At the present time when the curable treatment for AIDS has not been established, it is important to prevent further infection of HIV by spreading the proper knowledge on HIV and AIDS to many medical doctors, nurses, and other medical staffs as well as peoples. Medical staffs are needed to possess a better understanding of the disease status and the treatment. There is no need to have a overreaction to HIV infection and AIDS, since the route of infection and the source of infection are really restricted as in the case of hepatitis B or C virus. PMID- 8225170 TI - [A role of dentists in the management of HIV-infected persons]. AB - Oral manifestations of HIV infection include hairy leukoplakia, Sjogren's syndrome-like conditions, necrotizing stomatitis, Kaposi's sarcoma, and a variety of opportunistic viral or mycotic infections. The well awareness of HIV associated oral lesions can contribute to the early detection and diagnosis of HIV infection. Prevention and early treatment of common oral diseases including dental caries and periodontal disease, which may evolve into necrotizing gingivitis and stomatitis under immunodeficiency, is required to maintain quality of life and to improve prognosis of persons with AIDS. It is to be desired that many dentists take part in teams of the management of HIV-carrier. Another major issue is protection from HIV infection during dental treatment procedures. It is thought now that there is little probability of transmitting HIV infection during dental treatment. Current fears regarding virus-borne disease including hepatitis B and AIDS have prompted changes in dentistry's approach to infection control. Fear, ignorance and uncertainty, however, produce irrational behavior of dental staffs and persons without HIV infection toward patients with HIV infection. It is expected that the general public as well as dental staffs has a satisfactory knowledge about HIV infection. PMID- 8225171 TI - [Changes in epileptiform activities during sleep and sleep structures in temporal lobe epilepsy]. AB - We investigated the distribution of epileptiform activities (EA) and ictal discharges, and sleep characteristics in 23 patients with temporal lobe epilepsy (TLE) using polysomnography (PSG) for two consecutive nights. The cassette EEG system devised by our group was employed throughout the study to minimize th untoward effects of overnight recording. The patients consisted of nine men and fourteen women, aged from 17 to 60. Fourteen of them had been taking antiepileptic drugs for long periods of time. The remaining 9 were the untreated patients and examined before and after the commencement of drug therapy. Sleep characteristics of patients were compared to those of age and sex matched healty subjects. The following findings were obtained: 1) EA were found more often during NREM sleep than REM sleep in both treated and untreated patients. The frequency of EA increased in stage 3 + 4 compared to stage 1 or 2. 2) Changes of EA numbers during the consecutive sleep cycles were investigated. Twelve of 14 patients in the treated group showed curves which had decreasing tendency throughout the night. 3) Ictal discharges were observed exclusively in the stage REM or within 10 minutes before and after the REM stage. These phenomena were recognized in both treated and untreated groups. 4) Sleep characteristics of both treated and untreated patients indicated significantly decreased sleep efficiency and increase of % awakening. Moreover, % stage 4 significantly increased in untreated patients compared to healthy subjects. These findings were discussed in terms of the clinical characteristics with TLE and antiepileptic drugs on epileptic activities during sleep as well as sleep structures. PMID- 8225172 TI - [Effect of left ventricular assist device (LVAD) after myocardial infarction: study of reinfarction model]. AB - The effect of the left ventricular assist device (LVAD) after myocardial infarction was studied in 10 mongrel dogs. In the first experiment, the left anterior descending coronary artery (LAD) was occluded for one hour, then the LAD was reperfused. Two weeks later, the dogs were divided into two groups. In group I (control, n = 5), the LAD was occluded for 120 minutes, but the LVAD, implanted between left atrium and aorta, was not driven. In group II (pumping, n = 5), the LVAD was started 15 minutes after LAD occlusion, and continued for 115 minutes, then reperfused after 120 minutes of occlusion. The left ventricular segment length was monitored continuously by a pair of titanate-zirconate piezoelectric crystal. End-diastolic length (EDL) and end-systolic length (ESL) were identified. To describe segmental motion, % shortening was defined as follows: %shortening = (EDL-ESL)/EDL x 100. Just after sacrifice, the hearts were removed and sectioned transversely. Area of infarction was measured by planimetry as identified by incubation of heart slices in nitroblue tetrazolium dye. After LAD occlusion, % shortening decreased significantly in both groups. After reperfusion, % shortening increased, but was significantly low in the group I at 30 minutes of reperfusion.(p < 0.05). The EDL of ischemic region increased after coronary occlusion and remained high during LAD occlusion. The EDL was almost the same in both groups at 30 minutes of reperfusion. Area of infarction evaluated weight ratio and transmurality were 10.9 +/- 3.9% versus 11.3 +/- 1.4% and 81.2 +/- 8.9% versus 61.0 +/- 5.1% in comparison of group I versus group II.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225173 TI - [Effects of platelet aggregating inhibitor on pulmonary metastases of tumor cells after surgical resection]. AB - DN-9693, c-AMP: phosphodiesterase inhibits platelet aggregation induced by metastasizing tumor cells and blood-borne metastases of these tumors. Effects of this drug on pulmonary metastases was studied in wKA rats, which were sc implanted with 4-dimethylaminoazobenzene (DAB) induced KDH-8 tumor cells. KDH-8 cells (10(5)) were sc inoculated on day 0 and excised on day 20. DN-9693 was ip injected at a dose of 150 micrograms twice a day for 7 days pre operatively (-7 - 0) or perioperatively (-3 - +3) or postoperatively (0 - +7). The rats were sacrificed on day 20 after surgery, and lung weight and the number of surface pulmonary nodules were measured. Both were significantly decreased in the group of perioperative and postoperative administration of DN-9693. The survival of these rats were furthermore prolonged when Cyclophosphamide (40 mg/kg) was sc injected 3 days after surgical resection. KDH-8 tumor cells (10(4)) were iv inoculated on day 0, and DN-9693 was ip injected at a dose of 150 micrograms twice a day for 7 days on day 0 approximately 7. Rats were sacrificed on day 20, and same studies as above were done. In this artificial pulmonary metastases, the decrease of the number of lung nodules was observed in WKA rat treated with DN 9693. Platelet aggregation induced by KDH-8 tumor cells was inhibited by ADP inhibitor (apyrase, CP/CPK) and thrombin inhibitor (heparin, MD-805); KDH-8 tumor cells induced platelet aggregation by two different mechanisms: ADP-mediated aggregation and thrombin-mediated aggregation. This platelet aggregation by KDH-8 tumor cells was inhibited by DN-9693 with dose-dependency. DN-9693 had no direct anti-tumor effects either in vivo or in vitro. The results indicates that this drug prevents pulmonary metastases by inhibiting platelet aggregation. PMID- 8225174 TI - [Influences of cyclosporin A on wound healing]. AB - We examine influences of immunosuppression on wound healing, using C57BL/6 mice by carrageenan, anti-asialo GM 1 antibody and Cyclosporin A (CSA). Mice underwent a standardized dorsal skin incision and subcutaneous implantation of sterile polyvinyl alcohol sponge under ether anesthesia. They were sacrificed on the 14th and 21st day after wounding. The pelts excised, and fresh breaking strength was measured with a constant-speed tensiometer. The sponges were removed, and hydroxyproline was measured by Woessner's method. 1) Sixteen mice were divided into 2 groups, eight mice intraperitoneally received carrageenan to suppress macrophage function twice before and after wounding, and eight mice, as a control, received saline in the same way. The breaking strength in the carrageenan-treated mice was weaker than in the control on the 14th day and 21st day. Hydroxyproline content was not different between the treated group and control. 2) Sixteen mice were divided into 2 groups, eight mice intraperitoneally received anti-asialo GM 1 antibody to suppress natural killer cell function twice before and after wounding, and eight mice, as a control, received saline in the same way. The breaking strength in the anti-asialo GM 1 antibody treated mice was not significantly different from that in the control on the 14th or 21st day. Hydroxyproline content was not different in the treated group and control. 3) Thirty two mice were divided into 4 groups, eight mice intraperitoneally received CSA at a dose of 1.0 mg/kg twice before and after wounding, eight mice received CSA at a dose of 10.0 mg/body, eight mice received CSA at a dose of 100.0 mg/body to suppress T helper lymphocyte function, and eight mice, as a control, received Intralipos at a dose of 1.0 g/kg in the same way. The breaking strength in the 10.0 mg/kg and 100.0 mg/kg CSA-treated mice were weaker than in the control on the 14th day. There was no significant difference on the 21st day. The breaking strength in the 1.0 mg/kg treated mice was not different from in the control on the 14th day or 21st day. Histologically, there was no difference between CSA treated group and control. 4) Forty mice were divided into 4 groups, twenty mice received CSA at a dose of 40.0 mg/kg for once a day for 7 days before and 14 days after wounding respectively, twenty mice received Intralipos at a dose of 1.0 g/kg in the same way.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8225175 TI - [Intracellular mechanism of potentiating interaction of histaminergic and muscarinic receptors in rabbit parietal cells]. AB - Intracellular mechanisms of receptor-mediated acid secretion and potentiating interactions of receptors were explored in isolated gastric glands obtained from the stomach of New Zealand White rabbits. Intracellular free calcium concentration ([Ca2+]i) was measured by a digitized video-image analysis with a dual-wavelength excitation microspectrofluorometry in fura-2 loaded parietal cells, and acid secretion was determined in parallel experiments by monitoring the accumulation of [14C] aminopyrine (AP) in suspension of isolated glands. Histamine and carbachol stimulated acid secretion and increased [Ca2+]i in a concentration-dependent manner. The concentration-response curve for the acid producing effect of carbachol was overlapped by that for the [Ca2+]i-increasing effect. However, the concentration-response curve for the [Ca2+]i-increasing effect of histamine was shifted by approximately one order from that for the acid producing effect, indicating that a low concentration of histamine can also stimulate acid secretion without [Ca2+]i increase in the rabbit parietal cells. Cimetidine, a H2-blocker, inhibited the histamine-induced acid secretion and [Ca2+]i increase competitively, and those responses induced by carbachol non competitively. Carbachol at a concentration of 10(-3)M did not affect the histamine concentration of solution containing isolated gastric glands. Neither 10(-6)M 3-isobutyl-1-methyl-xanthine (IBMX) nor 3 x 10(-5)M dibutyryl adenosine 3',5'-cyclic monophosphate (dbcAMP) affected AP uptake and [Ca2+]i per se. However, these agents significantly potentiated the carbachol-induced acid secretion and [Ca2+]i increase. These results suggest that tonic stimulation of H2-receptors by a small amount of endogenous histamine potentiates the cholinergic acid secretion and [Ca2+]i increase. Parietal cells preconditioned by a small increase in intracellular cAMP may easily increase [Ca2+]i and acid secretion in response to cholinergic stimulation. PMID- 8225176 TI - [Immunochemical determination of advanced glycation end products in erythrocyte peripheral-membrane proteins from diabetic patients]. AB - Nonenzymatic glycation of protein leads to changes in protein structure and function, including browning, crosslinking and polymerization. There are several functional abnormalities in erythrocytes of diabetic patients. However, nothing definite is known about the relationship between these abnormalities and nonenzymatic glycation products, especially advanced glycation end products (AGEs). In this view, the present study was aimed to determine the amount of AGEs in diabetic erythrocyte peripheral-membrane proteins (EPMPs) by an ELISA system using AGE-specific antibodies. Erythrocyte membrane proteins from 19 control and 48 diabetic subjects were treated with 0.1 N-NaOH solution and then, the supernatants of each sample, containing EPMPs, were used to measure the amount of AGEs. The amount of AGEs in EPMPs was approximately 3 times greater in diabetic patients than in controls (70.7 +/- 51.8 AU vs. 22.5 +/- 7.7 AU (mean +/- SD), p < 0.001). There was a significant correlation between levels of AGEs in EPMPs and HbA1C (r = 0.779, p < 0.001). In SDS-PAGE analysis of EPMPs, two major bands, 221 kDa and 242 kDa were observed. These two bands were defined to be spectrin by immunoblotting with mouse monoclonal anti-human spectrin antibody. Based on these findings, it was speculated that the formation of AGEs in erythrocyte peripheral membrane proteins is related to the functional abnormalities of erythrocytes, including reduced deformability and lowered membrane fluidity, frequently observed in diabetic patients. PMID- 8225177 TI - [Effects of acidosis on the neuronal function following oxygen-glucose deprivation in the rat hippocampal slices]. AB - This study was designed to examine 1) whether cerebral ischemic damage is aggravated by accompanying acidosis and 2) which has more potential to cause neural damage between respiratory and metabolic acidosis. To investigate these points, inhibition and recovery of hippocampal evoked potentials were studied in vitro with different pH solutions. Population spike (PS) activity was recorded from CA 1 region after stimulation of the Schaffer collaterals of the 400 microns hippocampal slices from young Wister rats. Ischemic insult was mimicked by combined oxygen and glucose deprivation (OGD) of the perfusate for 15 min. PS activity was almost abolished by OGD in the test solutions, the pH of which was adjusted at either 7.4, 6.5, 6.0, 5.5, 5.0 or 4.5. PS persisted (though markedly depressed to 31.5% of control) with pH 6.5 solution, indicating that mild acidosis had antagonistic effects on ischemic injury. The recovery of PS in the control solution was significantly inhibited for the slices tested with pH 4.5 and 5.0 solutions. The results suggest that acidosis had diverse effects of ischemic cerebral damage. When 15 mM lactate was added to the solution, the recovery of PS was significantly inhibited for the slices tested with pH 5.5 or lower. When 30 mM lactate was added, the critical pH point at which the recovery of population spike was inhibited changed to 6.0, suggesting that ischemic cerebral damage was enhanced by lactate in a dose dependent fashion. By addition of CO2, the recovery of PS following OGD was inhibited to a similar degree as with 30 mM lactic acidosis. It is concluded that concomitant acidosis has divergent effects on the cerebral ischemic damage depending on the pH; mild acidosis had a protective effect but profound acidosis had an aggravating effect, the cross over point being at around pH 6.0. Lactate and CO2 potentiated the aggravating effects of acidosis even though the extracellular pH remained the same. PMID- 8225178 TI - [Studies on phosphorylation of estrogen receptor from porcine uterus]. AB - Estrogen receptor (ER) complex with estrogen interacts with estrogen responsive element (ERE) in target gene promoters and modulates transcription. ER phosphorylation process might be involved in the transcription regulation. To investigate the ER phosphorylation, the author have purified ER from porcine uterine cytosol through heparin-Sepharose column chromatography followed by immunoprecipitation with monoclonal anti ER antibody and protein A-Sepharose. The ER was phosphorylated with [gamma-32P] ATP without any protein kinase, showing that ER has autophosphorylation activity. 63kDa and 120kDa proteins were mainly detected by SDS-PAGE analysis following autoradiography. ER became phosphorylated in the presence of divalent cation such as Mg2+ or Ca2+ in a time-dependent manner and an ATP concentration-dependent manner. Phosphoamino acid analysis showed phosphorylation of serine residue. Phosphorylation was stimulated by estrogen (E2), and phosphorylated receptor was bound to DNA fragment that contained EREs. It is suggested that ER phosphorylation process might participate in transcription regulation. PMID- 8225179 TI - [Energy metabolism and work capacity of skeletal muscles in patients with chronic heart disease: a study using 31P-magnetic resonance spectroscopy]. AB - Limitation on muscle work capacity independent of blood supply has been postulated to be a contributing factor for a decrease in exercise tolerance in patients with chronic heart disease. The purpose of this study was to assess the exercise energy metabolism of skeletal muscles in chronic heart disease in relation to work capacity using 31-Phosphorus magnetic resonance spectroscopy (31P-MRS). Ten patients with chronic heart disease (group C) and 11 healthy subjects (group N) were studied. Peak oxygen uptake was determined during upright ergometer exercise. For the MRS study, the maximal cross-sectional area (MCA) of flexor muscles in the forearm and calf was first determined from MRI. Next, wrist and planter flexion exercises were performed separately while MRS spectra obtained. The exercise work load was applied according to each MCA (1J/cm2/min). Body weight and MCA were not significantly different between N and C. Peak oxygen uptake (ml/kg/min) and work rate (watt) during ergometer exercise ware significantly decreased in C (31 +/- 6 vs. 18 +/- 8 and 154 +/- 39 vs. 100 +/- 17, respectively, mean +/- SD, N vs. C, P < 0.05). In the MRS study, the decrease in pH was significantly greater in the forearm than in the calf in both groups. In comparing C with N, a decrease in PCr (suggestive of increased ADP) and a decrease in pH were greater in C in the calf. However, the peak work rates of both flexors were not significantly different between N and C. These results suggest that chronic heart disease probably affects the energy metabolism of mitochondria in skeletal muscles of the lower extremity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225180 TI - [Release of endothelium-derived relaxing factor (EDRF) from cultured vascular endothelial cells depends on extra- and intracellular Ca2+ concentrations]. AB - The objective of this study was to elucidate the role of extracellular and intracellular Ca2+ in EDRF release from vascular endothelial cells. Cultured fetal bovine aortic endothelial cells were stimulated by adenosine triphosphate (ATP), calcium ionophore (A23187), bradykinin (BKN) and acetylcholine (ACh). The amount of EDRF released was determined on the basis of the relaxation of rabbit aortic ring and the cGMP content of cultured smooth muscle cells derived from fetal bovine aorta. Changes in intracellular Ca2+ concentration ([Ca2+]i) were measured by a photometric fluorescence microscopy using Fura-2. ATP and A23187 induced dose-dependent increases in both [Ca2+]i and EDRF release. BKN increased both [Ca2+]i and EDRF release upon initial exposure (10(-8) M), but there were no further changes at higher concentrations because of desensitization. ACh induced no significant changes in either [Ca2+]i or EDRF release. There was a statistically significant correlation between agonist-induced changes in [Ca2+]i and the amount of EDRF released (p < 0.01). Removal of extracellular Ca2+ eliminated the continuous elevation induced by agonists of both [Ca2+]i and the amount of EDRF. The agonist-induced EDRF release was inhibited by L-NMA (N omega methyl L-arginine). The EDRF release reduced by L-NMA was restored by adding L arginine. Thus, the EDRF released from endothelial cells is thought to be an L arginine-derived nitric oxide (NO) or an NO-related substances. These findings suggest that [Ca2+]i is closely linked to the amount of EDRF released, and that extracellular Ca2+ is essential for EDRF release because its influx is involved in the continuous elevation of [Ca2+]i. PMID- 8225181 TI - [Functional studies of adhesion molecules on CD4-CD8- double negative T cells of autoimmune MRL/Mp-lpr/mice]. AB - MRL/Mp-lpr/lpr (MRL-lpr) mice have been used for a model of human systemic lupus erythematosus. This strain of mice homozygous for an autosomal recessive mutation, lpr (lymphoproliferation), develops massive lymphadenopathy with the expansion of CD4-CD8- (double negative; DN) T cells. Recently it was demonstrated that lpr mice have defects in the gene of Fas antigen which mediates apoptosis, indicating a possibility of defect in negative selection of autoreactive T cells in the thymus of lpr mice. However, the mechanisms that control the accumulation of DN T cells in lymph nodes, and the involvement of DN T cells in the clinical manifestation of disease, have not been well understood. In this study, the expression of various cell adhesion molecules on lymphocytes from MRL-lpr mice was examined. The strong expression of CD44 antigen as well as heat stable antigen (HSA) on abnormal DN T cells of lymph nodes was characteristic in MRL-lpr mice. Furthermore, the accumulation of DN T cells in lymph nodes might result from augmented binding of lymphocytes to endothelial cell surface of lymph nodes, possibly due to the failure of Mel-14 antigen shedding from DN T cell surface. In addition, it was found that monoclonal antibodies reactive with cell adhesion molecules such as CD44, Mel-14, CD45R and HSA expressed on DN T cells, could trigger the lytic activity of DN T cells and redirected DN T cell-mediated lysis of Fc-receptor-positive target cells (EL-4). In contrast to T cell receptor (TCR) mediated cytotoxicity, this redirected cytotoxicity was not inhibited by anti lymphocyte function associated antigen-1 (LFA-1) antibody. Thus, cell adhesion molecules may play a major role in delivering the transmembrane signal to DN T cells of MRL-lpr mice that trigger the lytic activity. It is likely that DN T cells of MRL-lpr mice induce tissue damages by the interaction with ligand on vascular endothelium or extracellular matrix in vivo. PMID- 8225182 TI - [An experimental study on absorbable and non-absorbable suture materials in aortic anastomoses: influence of the growth to suture lines and anastomotic sites]. AB - Ent-to-end anastomoses of the abdominal aorta in dogs and growing pigs were performed with absorbable sutures and non-absorbable sutures. Comparison studies between the two suture materials were done with respect to mechanical properties, angiographic findings, and macroscopic and microscopic examinations. In macroscopic examinations, animals with non-absorbable suture materials developed thickening at the site of the anastomosis and showed protrusion of the suture materials into the intraluminal space of the aorta with growing pigs. However, those findings were not observed in the animals with absorbable suture materials. In histological examinations, prolonged inflammatory reaction around the anastomotic site was observed in the non-absorbable suture group, but excellent healing without any inflammatory reaction was a constant finding in the absorbable suture group. Animals with the absorbable suture materials showed good mechanical properties evaluating the compliance and tensile strength, compared to that of animals with the non-absorbable suture materials. Growth of a aortic anastomosis after application of absorbable suture material seems to be possible without any complication and can be recommended as adequate suture materials for application in young growing humans. PMID- 8225183 TI - Enhancement of DNA polymerase beta activity in the pituitary gland by hormonal feedback. AB - The activity of DNA polymerase beta (beta-enzyme) in bovine and rat pituitary glands was remarkably higher in the anterior than in the posterior lobe. In sucrose gradient centrifugation, the activity in the anterior lobe of the bovine pituitary gland appeared at the sedimentation constant, 3.3 S, while that of rat pituitary gland appeared at 3.3 S, 7.3 S and 12 S, respectively. The activity in each fraction increased in the anterior lobes of rat pituitaries that were stimulated to release TSH by decreased thyroid hormone levels induced by an antithyroid drug. These results suggest that the activity of the beta-enzyme is high in the cells of the anterior lobe which are hyperfunctioned to produce pituitary thyreotrophic hormone and may be controlled by hormonal feedback regulation. PMID- 8225184 TI - Influence of sodium and potassium diets on adrenal vasopressin content and direct effects of vasopressin on aldosterone synthesis in adrenocortical cells. AB - Two questions were investigated to elucidate the adrenal function of arginine vasopressin (AVP) in controlling aldosterone secretion: Are vasopressin tissue concentrations in the adrenal modulated by diets that influence aldosterone secretion? Does its direct stimulation of aldosterone secretion from adrenal cells in vitro suggest that vasopressin plays an important role in controlling aldosterone? Diets that modulate aldosterone secretion from rat adrenals did not modulate concentrations of AVP in this tissue: Adrenal AVP concentrations did not differ significantly between rats on low- and high-sodium (22.9 +/- 3.8 and 32.4 +/- 6.4 pg/g) or between low- and high-potassium (25.6 +/- 2.5 and 12.5 +/- 7.3 pg/g) diets. Although the existence of AVP receptors on adrenocortical cells incubated in vitro and the stability of AVP during cell incubation was confirmed, AVP had only a minor direct effect on steroid secretion in short-term incubations alone and in combination with angiotensin II (A II), potassium and atrial natriuretic factor (ANF). The importance of AVP for an indirect control of aldosterone secretion in vivo is discussed. PMID- 8225185 TI - Parathyroid hormone related peptide (1-34) and 1,25-dihydroxyvitamin D, have no additive effects on bone tartrate resistant acid phosphatase activity in fetal rat calvaria cultures. AB - Parathyroid hormone related peptide (PTHrP) and 1,25-dihydroxyvitamin D are known to be resorptive agents which could contribute to the development of hypercalcemia in humoral hypercalcemia of malignancy (HHM) syndrome in Walker 256 tumor bearing rats. In order to clarify some aspects about the relative contribution of these factors to bone resorption, we have determined the effects produced by PTHrP (1-34) and/or 1,25(OH)2D3 on tartrate resistant acid phosphatase (TRAP), a biochemical marker of bone resorption, in fetal rat calvaria cultures. At the same time, bovine parathyroid hormone (PTH) (1-34) was used as a control in the experiments in order to compare its effects with those produced by the other two agonists. In the present work, 10(-7) M PTH (1-34), 10( 7) M PTHrP (1-34) and 10(-8) M or 10(-10) M 1,25(OH)2D3 produced a significant increase in TRAP activity, when these agonists were added to the calvaria culture. Surprisingly, and in spite of the different ways of action of PTH, PTHrP and 1,25(OH)2D3, their actions are not additive in our experiment. The results of the present work suggest that any of the two implicated factors PTHrP or 1,25(OH)2D3 could be individually responsible for the high rate of bone resorption that takes place in HHM syndrome in Walker 256 carcinosarcoma bearing rats, although other different agents, like TGF, could also be implicated. PMID- 8225186 TI - Role of dual signal transduction systems in the stimulation of bone resorption by parathyroid hormone-related peptide. The direct involvement of cAMP-dependent protein kinase. AB - The present study was performed to compare the effect of parathyroid hormone related peptide (PTHrP) on bone resorption with that of parathyroid hormone (PTH) and clarify the participation of PTHrP-responsive dual signal transduction systems involving cAMP-dependent protein kinase (PKA) and calcium/protein kinase C (Ca/PKC) in the stimulation of bone resorption by PTHrP. Bone resorbing activity was estimated as the number of pits formed on the dentine slice and total area of pits per slice in bone cells derived from 2 week-old mice. Human (h)PTHrP-(1-34) (10(7) M) stimulated bone resorption as potent as hPTH-(1-34) (10(7) M) did. The stimulation of bone resorption by hPTHrP-(1-34) and hPTH-(1 34) was equally blocked by either simultaneous treatment with 10(-8) M Elcatonin (eel calcitonin derivative; from Asahi Chemical Industry, Tokyo, Japan) [corrected] or pretreatment with 10(-7) M [Nle8,18Tyr34]hPTH-(3-34)amide. Rp cAMPs, an antagonist in the activation of PKA, equally attenuated bone resorption stimulated by PTHrP as well as by PTH. A23187 (10(-7) M) caused a significant stimulation of bone resorption. These findings indicate the direct involvement of PKA activation and a contributory role of an increase in cytosolic calcium in the stimulation of bone resorption by PTHrP and suggest that PTHrP stimulates bone resorption presumably through the same mechanism as PTH does. PMID- 8225187 TI - Growth hormone and prolactin variants in normal subjects. Relative proportions in morning and afternoon samples. AB - There are multiple molecular forms of both growth hormone (GH) and prolactin (PRL). Traditionally the two hormones have been measured by radioimmunoassay (RIA) techniques. Recently, several molecular variants of these hormones have been discovered using Western blotting techniques: four GH size variants, 27K GH, 22K GH (the classical form), 20K GH (an alternatively-spliced form), and 17K GH, and two PRL structural variants, a glycosylated (G-PRL) and a nonglycosylated form. In this study, we measured these GH and PRL variants in 18 normal subjects in the morning in a fasting state and in the afternoon in a non-fasting state. Contrary to expectations, the predominant serum GH form in both morning and afternoon samples was found to be 17K, not 22K GH, accounting for 82-89% of the total circulating GH. The predominant serum PRL form was found to be the nonglycosylated variant, constituting 83-84% of the total circulating PRL. None of the GH or PRL variants were significantly different when comparing morning to afternoon samples. These results provide, for the first time, evidence for the existence of two new GH-immunoreactive components in human sera, the 17K and 27K GH, the former in proportions often higher than those of the classical 22K GH, and argue for the need to measure them individually. PMID- 8225188 TI - Evaluation of thyrotropin secretion before and after TRH by third generation chemiluminescent assay. Assessment of subclinical hyperthyroidism. AB - The recent introduction of third generation assays for TSH has led to a considerable improvement of assay sensitivity. To assess the clinical significance of subnormal basal TSH (b-TSH) values (< 0.2 microU/ml), we investigated b-TSH and TRH-stimulated TSH (r-TSH) by means of a new, highly sensitive immunochemiluminometric assay in 105 euthyroid subjects, 45 patients with overt hyperthyroidism and 18 patients suspected of having subclinical hyperthyroidism. A weak, albeit statistically significant, correlation (r = 0.48) was found between b-TSH and r-TSH and also between b-TSH and delta-TSH (r = 0.31) in euthyroid subjects. Consideration of b-TSH alone correctly identified 90 % of euthyroid subjects in this group; 10 of 105 apparently euthyroid subjects presented delta-TSH suggesting subclinical hyperthyroidism. While b-TSH was detectable (> 0.04 microU/ml) in 8 of 45 (18%) of hyperthyroid patients, all (100%) were abnormal in both b-TSH and r-TSH. 14 of 18 (78%) of patients with subclinical hyperthyroidism exhibited a blunted TSH response to stimulation (delta-TSH < 2 microU/ml). These results suggest that although the new generation of TSH assays can be a valuable addition to the diagnostic arsenal of thyroid function tests, certain limitations must still be accepted. Specifically, b-TSH in the "grey zone" (0.1-0.2 microU/ml) appears to be a less than reliable predictor of thyroid function. PMID- 8225189 TI - Melatonin response to TRH in prepubertal and pubertal healthy subjects. AB - The current study was carried out to evaluate the influence of thyrotrophin releasing hormone (TRH) on melatonin secretion in healthy subjects. The study included 31 subjects of both sexes (prepubertal subjects: 15; pubertal subjects: 16). They were treated with TRH (0.2 mg i.v. as a bolus) with blood samples being collected at 0, 20, 60, 120, 180 min. after hormone administration; the results were compared with those of subjects infused with saline only. Serum levels of melatonin were measured with a double antibody RIA method. A significant increase in melatonin levels after TRH was seen only in the prepubertal females, with a peak at 120 min. On the contrary, no significant melatonin changes were seen in the prepubertal males or in either pubertal males and females. These results suggest the existence of sex- and age-related differences in melatonin responses to TRH in humans. PMID- 8225190 TI - Increase in skeletal muscle blood flow but not in renal blood flow during euglycemic hyperinsulinemia in man. AB - In order to investigate the effect of euglycemic hyperinsulinemia on skeletal muscle blood flow and renal blood flow, catheters were inserted into both femoral arteries, one femoral vein and one renal vein of 7 healthy men. Constant infusions of indocyanine-green dye (intraarterial) and of p-aminohippuric acid (intra-venous) were used to estimate leg plasma flow (ELPF) and renal blood flow (ERPF), respectively, prior to and during a euglycemic, hyperinsulinemic clamp (1.0 mU/kg.min of human insulin, serum concentrations of insulin before and during the clamp: 4.6 +/- 0.9 microU/ml and 65.5 +/- 20.6 microU/ml, respectively, t = 120 min). ERPF (basal: 1220 +/- 320 ml/min) remained unchanged throughout the period of induced hyperinsulinemia in each volunteer (mean: 1135 +/- 490 ml/min), whereas mean leg plasma flow (ELPF) rose from basal 206 +/- 99 ml/min up to 275 ml/min 90 minutes after the beginning of the euglycemic clamp study (p < 0.01). This was due to the marked rise in ELPF from 149 +/- 24 ml/min up to 243 +/- 25 ml/min (p < 0.01) seen in 5 subjects. In two men, who presented a markedly higher basal ELPF (332 and 365 ml/min, respectively), no further rise in ELPF was seen during induced hyperinsulinemia. Fractional renal extraction of insulin was unchanged during induced hyperinsulinemia (28 +/- 5%; basal: 22 +/- 18%), as was fractional extraction of insulin by the leg (10 +/- 5%; basal: 13 +/ 11%). The observed dissociation of ERPF and ELPF suggests a differential response to insulin in renal vs. leg vasculature which possibly is due to increased peripheral glucose metabolism. PMID- 8225191 TI - Twenty-four hours hormone secretion profiles of cortisol and estradiol in preterm labour. AB - We investigated the 24 hour hormone secretion profiles in women with preterm labour. Although estradiol secretion seems to be unaffected, we found an variable estradiol-cortisol-shift due to a cortisol increase. Cortisol circadian secretion persists in preterm labour, but maximum secretion is moved to midday. Further investigations are necessary to study the basic mechanisms underlying this process. PMID- 8225192 TI - Lipid peroxide and glutathione levels in the liver and its subcellular fractions of alloxan diabetic rats. PMID- 8225193 TI - Lipid bound sialic acid concentration in mice with myeloid leukemia and alloxan diabetes. PMID- 8225194 TI - Haemostatic mechanisms are normal despite increased vascular mortality in hypopituitary adults. PMID- 8225195 TI - Increases in plasma prostaglandin E2 after eccentric exercise. A preliminary report. PMID- 8225196 TI - HLA family study in a group of Greek women with non-classical 21-hydroxylase deficiency. PMID- 8225197 TI - Metabolic derangements in excessive insulin and sulfonylurea therapy. AB - Mild streptozotocin diabetic rats, characterized by normal or slightly elevated fasting blood glucose levels and glucose intolerance, treated with excessive doses of monocomponent pork insulin (0.5 U/day) (I) or glybenclamide (0.6 mg/day) (S) were compared to controls (C) and streptozotocin-diabetic rats without treatment (D). Intravenous glucose tolerance tests (0.75 g/kg) were performed in all animals and repeated after overtreatment. Insulin binding and insulin-induced D-(U-14C)-glucose transport and oxidation were also determined in isolated epididymal adipocytes. Diabetic rats showed a failure in the initial phase of insulin release and glucose intolerance as compared with (C). In overtreated rats glucose tolerance worsened (p < 0.05) after therapy. Maximal insulin binding by isolated adipocytes at tracer insulin concentration was unchanged after excessive insulin or sulfonylurea therapy. Besides, glucose transport and oxidation in the cells of overtreated rats were greater than in D and even greater than in C. These apparently divergent results, i.e. deterioration of glucose tolerance with increased insulin action in adipocytes suggest that overtreatment induces a state of resistance to hormone action in other target tissue(s) than the adipose one, possibly muscle. PMID- 8225198 TI - Decreased glucose transporter (GLUT 4) content in insulin-sensitive tissues of obese aurothioglucose- and monosodium glutamate-treated mice. AB - We assessed the content of two isoforms of glucose transporter (GLUT 1 and GLUT 4) in insulin-sensitive tissues of hypothalamic obese mice treated with either aurothioglucose (AuTG) or monosodium glutamate (MSG). The animals were studied when obesity had reached a plateau, and they were clearly insulin resistant. We studied different membrane fractions from white adipose tissue (WAT), such as fat free extract (FFE), plasma membrane (PM) and microsomal (M) fractions. GLUT 4 expressed per protein content displayed a decrease of 50% (p < 0.001) in all membrane fractions of AuTG- and MSG-treated mice. The PM GLUT 4 content, expressed per cell surface area, was reduced by 70% (p < 0.001) in obese mice, and the total FFE GLUT 4, expressed per total fat mass, was 5 times reduced in obese mice. Compared to control mice, obese mice showed a reduction (p < 0.01) of the GLUT 4 amount by 30% (AuTG) and by 40% (MSG) in total membrane fraction (TM) of skeletal and cardiac muscles. Similarly, a reduction of the GLUT 4 amount by 40% (AuTG) and by 45% (MSG) in FFE of brown adipose tissue was observed. The GLUT 1 content in FFE of WAT and TM of skeletal muscle showed no significant difference among the different animal groups. These results confirm a decreased expression of GLUT 4, but not of GLUT 1, in insulin-sensitive tissues, which may contribute to the impaired glucose utilization in these obese animals. PMID- 8225199 TI - Calcitonin gene regulation in vivo. AB - In view of the importance of the caliotropic hormones to calcium homeostasis and bone strength we have studied the regulation of the expression of the calcitonin gene in vivo in the rat. Normal rats were injected with 1,25-dihydroxyvitamin D3 (12.5-200 pmol/100 g body weight) and calcitonin mRNA levels measured. 1,25(OH)2D3 led to a marked decrease in calcitonin mRNA levels, which reached 4% of basal at 24 h. Nuclear transcript experiments showed that the effect was transcriptional. After large changes in serum calcium for periods from 6 h to 3 weeks there were no differences in calcitonin mRNA levels. Estrogens given to ovariectomized rats led to increases in calcitonin mRNA. The estrogen receptor was present in the thyroid C-cells. Therefore the calcitonin gene is regulated by vitamin D and estrogens, but not by calcium. PMID- 8225200 TI - Regulation of calcitonin secretion in vitro. AB - The concentration of extracellular calcium rightly regulates calcitonin secretion by calcium influx through dihydropyridine-sensitive voltage-dependent calcium channels; the result is an increase in intracellular calcium. There also exists a cAMP-dependent pathway of calcitonin release activated by glucagon or growth hormone releasing hormone. In thyroid C-cells, as in all cells, there is dual regulation of adenylate cyclase, mediated by inhibitory or stimulatory G proteins; glucagon stimulated cAMP production can be inhibited by somatostatin via pertussis toxin sensitive inhibitory G proteins. Somatostatin inhibits not only cAMP dependent but also calcium-dependent calcitonin secretion. Furthermore, somatostatin inhibits voltage dependent calcium channel currents thereby lowering cytosolic calcium. These actions also involve a pertussis toxin-sensitive inhibitory G protein but they occur independently of changes in the cytosolic cAMP concentration. Thus multiple interactions between second messenger systems at different cellular levels modulate calcitonin secretion. PMID- 8225201 TI - Characterization of the structural and functional properties of cloned calcitonin receptor cDNAs. AB - We have recently cloned CTRs from cDNA libraries prepared from porcine renal and human ovarian cell lines. In situ hybridization and Northern analysis confirm the widespread distribution of CTR mRNA in numerous tissues. Hydropathy plots of the predicted amino acid sequence of the receptors demonstrate multiple hydrophobic regions that could generate 7 transmembrane spanning domains, similar to other G protein-coupled receptors. Searches of databanks for proteins with related amino acid sequences reveals that the CTRs are closely related to the receptors for parathyroid hormone/parathyroid hormone related peptide, secretin, vasoactive intestinal peptide, growth hormone releasing hormone, glucagon-like peptide-1 and glucagon. These receptors have no significant sequence homology to other G protein-coupled receptors, and therefore, appear to comprise a distinct receptor family. Expression of the hCTR or pCTR in COS cells results in expression of high affinity CTRs which are coupled to adenylate cyclase (AC). The hCTR, however, demonstrates higher affinity for human and salmon CT compared to the pCTR. Both CTRs demonstrate low affinity binding and AC activation in response to calcitonin gene related peptide, amylin or secretin, providing a possible explanation for the cross-reactivity among these peptides in vivo. Stable transfectants expressing the pCTR increase cAMP levels and increases in cytosolic free Ca2+ concentration consistent with dual coupling to AC and phospholipase C. Additional studies will help to establish the structural basis for this functional property as well as the evolutionary relationship of the members of this newly identified family of receptors. PMID- 8225202 TI - Long-term treatment with calcitonin in osteoporosis. AB - Many studies have shown that calcitonin has beneficial effects on the clinical and biological disturbances of diseases characterized by an excessive bone remodelling. Several controlled clinical studies have shown that long-term calcitonin treatment, given by parenteral or intranasal routes, exerts a beneficial positive effect on bone mass and bone turnover in established postmenopausal osteoporosis. Calcitonin therapy is particularly indicated for patients with high turnover osteoporosis where results show a net gain of bone mineral in the axial skeleton and a slowing of bone loss in the appendicular bones. Recently, calcitonin was shown to induce a significant reduction in postmenopausal osteoporotic vertebral fractures. Due to receptor down-regulation a resistance to the hormone may occur after 12-18 months of continuous treatment. Reported results of long-term calcitonin treatment demonstrated that it is possible to delay or to avoid the "resistance" to calcitonin by the cyclical or discontinued administration of the hormone. PMID- 8225203 TI - Neutralizing antibodies against calcitonin. AB - The use of calcitonin (CT) is established as a treatment of Paget's disease of bone and postmenopausal osteoporosis (PMO). Salmon calcitonin (sCT), which differs in 14 of the 32 amino acids from human calcitonin, has found a wider distribution world wide, although antibody formation against sCT has been reported in more than 70% of the patients on continuous sCT treatment. The clinical significance of these antibodies has been discussed controversially, because the occurrence of antibodies is not always associated with the development of secondary resistance. Using an in vitro bioassay, based on the CT mediated increase of the cyclic AMP (cAMP) production of the human breast cancer cell line T 47 D we could identify a neutralizing activity against sCT in the serum of a subset of patients with formation of antibodies against sCT which was related to the development of secondary resistance. Antibody formation against human calcitonin (hCT) has been reported only once before. Binding and neutralizing antibodies were now observed in 1 of 33 patients with PMO treated with hCT. Due to a low neutralizing activity, clinical sequelae were not to be expected in this patient. The formation of neutralizing antibodies against calcitonin is common after treatment with salmon but a rare phenomenon after treatment with human calcitonin. We recommend monitoring of patients with postmenopausal osteoporosis and Paget's disease of bone on long term treatment with sCT or hCT for neutralizing antibody formation in order to evaluate the therapeutic effect of treatment. PMID- 8225204 TI - Effect of aminohydroxypropylidene diphosphonate on the bone metabolism of patients with parathyroid adenoma. AB - Aminohydroxypropylidene diphosphonate (APD), a potent inhibitor of bone resorption, is used to control hypercalcemia in various diseases. It is less effective, however, in the management of hypercalcemia induced by primary hyperparathyroidism. We investigated the effect of APD on the bone metabolism of five patients with parathyroid adenoma. Before parathyroidectomy, 30 mg of APD was administered intravenously. Serum calcium decreased in all cases one to two days after APD administration, although it did not decrease to the normal range. Serum phosphorus also decreased. Urine calcium and hydroxyproline excretion, markers of osteoclasts activity, decreased dramatically. Serum alkaline phosphatase (ALP) and osteocalcin, markers of osteoblast activity, decreased after APD administration. Serum intact parathyroid hormone (PTH) and 1,25 dihydroxy-vitamin D (1,25[OH]2D) increased. These results indicate that APD is partially effective in the management of preoperative serum calcium level in patients with parathyroid adenoma. As osteoclasts activity is inhibited by APD, osteoblasts activity is also suppressed. Elevation of PTH and 1,25(OH)2D after APD-induced decrease in serum calcium level may explain the partial and limited effect of APD on lowering serum calcium in patients with parathyroid adenoma. PMID- 8225205 TI - Twenty-four hours urinary hydroxyproline excretion in long-term experimental diabetes mellitus. PMID- 8225206 TI - Regulation of proliferation of bovine oviductal epithelial cells by estradiol. Interactions with progesterone, interferon-tau and interferon-alpha. PMID- 8225207 TI - Simvastatin influences testicular steroidogenesis in human. PMID- 8225208 TI - Southern-blot analysis and simultaneous in situ detection of hepatitis B virus associated DNA and antigens in patients with end-stage liver disease. AB - To gain new insights into the pathogenesis of hepatitis B virus-induced chronic liver disease, we have used nonisotopic in situ detection methods for the simultaneous analysis of hepatitis B virus DNA and antigens at the single-cell level. Paraffin-embedded liver specimens from 23 cirrhotic patients (12 HBsAg positive and 11 HBsAg negative) who underwent liver transplantation were evaluated by in situ hybridization with a digoxigenin-labeled DNA probe and digoxigenin detection system and by immunohistochemistry with an enhanced biotin streptavidin technique. DNAs extracted from liver and serum specimens were analyzed by Southern- and slot-blot hybridization, respectively. Using the in situ techniques, we detected hepatitis B virus-specific DNA and antigens in 11 of 12 HBsAg-positive patients and in none of the 11 HBsAg-negative individuals. Replicative intermediates of hepatitis B virus DNA were detected by Southern-blot analysis in the same 11 HBsAg-positive patients, 6 of whom had no serological markers of hepatitis B virus replication. Therefore a good correlation was found between the results obtained by the in situ and Southern-blot hybridization analyses of tissue specimens. However, a lack of correlation was found between serum- and tissue-associated markers of viral replication. In addition, the simultaneous in situ detection analyses revealed that some hepatocytes containing high levels of viral DNA were devoid of detectable HBcAg, suggesting a mechanism by which the virus may escape immunological surveillance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225209 TI - Hepatic and extrahepatic HCV RNA strands in chronic hepatitis C: different patterns of response to interferon treatment. AB - We investigated the presence of positive (genomic) and negative (replicative intermediate) hepatitis C virus RNA strands in liver, peripheral mononuclear cells and serum from patients with chronic hepatitis C using a selective and semiquantitative polymerase chain reaction procedure. Negative and positive hepatitis C virus RNA strands were present in liver, serum and lymphoid cells in all untreated patients and in all those who did not respond to interferon therapy. In the latter group of patients, the titers of RNA strands in the liver and peripheral mononuclear cells at the end of the treatment were similar to those encountered in untreated patients, but the serum titers were about 100 times lower than pretreatment values. In patients who responded to interferon with normalization of serum aminotransferase levels (n = 10), the rate of detection and the titer of the two viral strands in liver, serum and mononuclear cells were markedly decreased at the end of the therapy. In the six responders who did not relapse after interferon withdrawal, both hepatitis C virus RNA strands were absent from the liver, serum and lymphoid cells. By contrast, the positive RNA strand was present in liver cells, mononuclear cells or both at the end of therapy in all patients who experienced posttherapy relapse. In conclusion, our results indicate that interferon can clear hepatitis C virus from hepatic and extrahepatic sites only in responder patients. Disappearance of genomic hepatitis C virus RNA from the liver and from mononuclear cells may predict complete response without posttherapy relapse. PMID- 8225210 TI - Positron-emission tomographic localization of abnormalities of brain metabolism in patients with minimal hepatic encephalopathy. AB - Many patients with compensated cirrhosis without overt hepatic encephalopathy have deficits in visual-spatial perception, a condition we call minimal hepatic encephalopathy. Five patients with alcohol-induced cirrhosis and nine control subjects underwent positron-emission tomographic imaging of the brain with 18F fluorodeoxyglucose. Patients also underwent neuropsychological and clinical chemistry tests. The patients had mild arterial hyperammonemia (62 +/- 13 mumol/L, range = 11 to 35 mumol/L) and other abnormalities typical of patients with cirrhosis. The patients' mean percentile scores on the digit symbol and block design subtests, from the Wechsler Adult Intelligence Scale (revised), and Purdue pegboard test were 11 +/- 7, 24 +/- 7 and 7 +/- 8 (right hand). Tests of vocabulary, memory, and new learning were normal. The technique of statistical parametric mapping was used to identify regions where cerebral 18F fluorodeoxyglucose uptake and metabolism were abnormal. We noted significant reductions in the cingulate gyrus, a center mediating attention, target analysis and response formulation and significant increases in visual associative regions subserving motion and color perception and object orientation. We suggest that minimal hepatic encephalopathy is due to a deficit in the detection and formulation of responses to visual stimuli, a function of the cingulate, which is a part of the anterior attentional system of the brain. Increases in 18F fluorodeoxyglucose metabolism may be compensatory. These studies show that brain regions differ in their sensitivity to the agents that cause hepatic encephalopathy and that positron-emission tomography is useful in studying the pathophysiology of this disorder. PMID- 8225211 TI - Factors predictive of liver histopathological appearance in chronic alcoholic pancreatitis with common bile duct stenosis and increased serum alkaline phosphatase. AB - In the course of alcoholic chronic pancreatitis, increased serum alkaline phosphatase level is usually caused by common bile duct stenosis but may also be due to alcoholic liver disease. The aims of this prospective study were to investigate whether clinical, biochemical and radiological factors could predict liver histopathological appearance. The study comprised 48 patients with chronic alcoholic pancreatitis, common bile duct stenosis and increased serum alkaline phosphatase levels; clinical, biochemical, radiological and histological data were recorded in all cases. Liver biopsy examination (surgical [n = 45] or intercostal [n = 3]) showed (a) biliary obstructive liver abnormalities (n = 33), which were severe in 20 cases (biliary fibrosis in 15, secondary biliary cirrhosis in 3, secondary sclerosing cholangitis in 2) and moderate in 13 cases; (b) alcoholic liver disease in 9; and (c) normal liver in 6. Clinical, biochemical and radiological data were not statistically different between patients with biliary obstructive liver disease and those with alcoholic liver disease. Forty-five patients underwent surgery; two patients with alcoholic hepatitis died after surgery, at the beginning of this study. We conclude that in chronic alcoholic pancreatitis with common bile duct stenosis and increased serum alkaline phosphatase levels, clinical, biochemical and radiological data cannot be used to predict the type of liver lesions. Therefore liver biopsy is warranted to identify (a) alcoholic hepatitis, which increases operative risk; and (b) biliary obstructive liver disease, frequent and often severe, in which surgical biliary decompression should be considered. PMID- 8225212 TI - Reduction of variceal pressure by propranolol: comparison of the effects on portal pressure and azygos blood flow in patients with cirrhosis. AB - This study investigated the correlation between changes in hepatic hemodynamics and esophageal variceal pressure--measured with a noninvasive, pressure-sensitive endoscopic gauge--in 37 portal-hypertensive cirrhotic patients receiving propranolol (0.15 mg/kg, intravenously; n = 21) or placebo (n = 16) under strict double-blind conditions. Placebo administration had no effect on hepatic venous pressure gradient, azygos blood flow or variceal pressure. Propranolol caused a significant reduction in hepatic venous pressure gradient (from 19.6 +/- 1 to 17.3 +/- 1 mm Hg, p < 0.001), azygos blood flow (from 0.61 +/- 0.06 to 0.39 +/- 0.03 L/min, p < 0.001) and variceal pressure (from 13.1 +/- 0.9 to 10.2 +/- 0.9 mm Hg, p < 0.001). In eight patients (propranolol nonresponders) hepatic venous pressure gradient was not modified or decreased by less than 10% after propranolol (mean change, -4.1 +/- 1.6%). However, we found no significant differences between propranolol responders and nonresponders with regard to the decrease in variceal pressure (3.3 +/- 0.7 vs. 2.3 +/- 1.4 mm Hg) and azygos blood flow (0.23 +/- 0.07 vs. 0.21 +/- 0.07 L/min). As expected, in most propranolol responders (11 of 13), reduction in hepatic venous pressure gradient was accompanied by a similar response in variceal pressure (> 10% decrease). However, among propranolol nonresponders, in terms of reduction in hepatic venous pressure gradient, four out of eight patients had decreases greater than 10% in variceal pressure. The results of this study confirm that reduction in hepatic venous pressure gradient by propranolol is associated with a significant decrease in variceal pressure and azygos blood flow.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225213 TI - Resolution of liver biopsy alterations in three siblings with bile acid treatment of an inborn error of bile acid metabolism (delta 4-3-oxosteroid 5 beta-reductase deficiency). AB - Identical male twins and their brother, cholestatic from birth, with delta 4-3 oxosteroid 5 beta-reductase deficiency, were studied by serial liver biopsy. Spectrometry documented defective primary bile acid synthesis and markedly increased levels of atypical oxo and allo bile acids in urine and serum. Hepatocellular cholestasis and giant-cell transformation resolved in parallel with clinical and biochemical recovery during oral bile acid administration. In the twins, portal fibrosis stabilized at a mild level; they are well as 5-yr-olds at this writing. Follow-up biopsy in their brother at 8 mo was normal, and he is doing well at 3 yr of age. Hepatic ultrastructural alterations in all three were characterized by abnormalities of bile canaliculi including small bile plugs, diverticulae and latticelike elaborations of hepatocellular membranes adjacent to bile canaliculi that were shown to have resolved completely on subsequent biopsies. Eight additional cases have been detected on urine screening; only two of these patients have survived, on bile acid therapy. Early diagnosis and treatment improves the prognosis of this otherwise lethal inborn error of bile acid synthesis. PMID- 8225214 TI - Randomized controlled trial of desmopressin plus terlipressin vs. terlipressin alone for the treatment of acute variceal hemorrhage in cirrhotic patients: a multicenter, double-blind study. New Italian Endoscopic Club. AB - 1-Deamino-8-D-arginine vasopressin (DDAVP, desmopressin), a synthetic analog of the antidiuretic hormone L-arginine vasopressin, improves hemostasis parameters in cirrhotic patients. Hence its use in combination with a vasoactive drug such as terlipressin might improve the performance of this drug in controlling variceal bleeding. The aim of this trial was to compare the efficacy of desmopressin plus terlipressin with that of terlipressin alone in controlling acute variceal hemorrhage. Cirrhotic patients with active variceal hemorrhage diagnosed endoscopically were randomized within 2 hr of admission to receive desmopressin plus terlipressin or placebo plus terlipressin. Terlipressin (2 mg, intravenous bolus) was given at time 0 and every 4 hr thereafter for 24 hr. Desmopressin (0.3 microgram/kg, intravenously) or placebo was given in saline solution over 30 min at time 0 and at 26 hr. Patients were monitored for 24 hr after cessation of treatment. Treatment failure was defined as recurrence of active bleeding during treatment or within the 24 hr after treatment. After enrolling 51 of the planned 84 patients, we carried out an interim analysis. Treatment failure occurred in 13 of 24 patients randomized to receive desmopressin plus terlipressin (54.2%) and in 6 of 22 patients randomized to receive terlipressin (27.3%) (p = 0.06, Fisher's exact test). The trial was interrupted at this stage because patients treated with the "new" therapy fared worse than those treated with the standard therapy, and the possibility of reversing this trend by completing the trial was deemed remote. The addition of desmopressin does not improve and may worsen the efficacy of terlipressin in controlling acute variceal bleeding in cirrhotic patients. PMID- 8225215 TI - Steroid receptors and heat-shock proteins in patients with primary biliary cirrhosis. AB - Primary biliary cirrhosis has a definite female preponderance. Increased estrogen levels have been found in patients with this disease; however no studies indicate the status of sex hormone steroid receptors in primary biliary cirrhosis patients. In this study the occurrence and distribution of estrogen receptors, progesterone receptors and androgen receptors in liver biopsy specimens from patients with primary biliary cirrhosis were examined and compared with these receptors in the normal liver. In addition, three heat-shock proteins associated with steroid receptors (90 kD, 70 kD and 27 kD) were examined. All of the receptor proteins were detected on immunocytochemical study using specific receptor antibodies; monoclonal and polyclonal antibodies were also used to detect the heat-shock proteins. Normal bile duct epithelial cells displayed low to-moderate amount of estrogen receptors and abundant 90- kD, 70- kD and 27-kD heat-shock protein expression, whereas normal hepatocytes showed moderate estrogen receptor and 90-kD heat-shock protein and high 70-kD heat-shock protein expression. Expression of 70-kD heat-shock protein was due mainly to the constitutive form of this protein (hsc72). In patients with primary biliary cirrhosis, significant increases in estrogen receptor and 90-kD heat-shock protein content were seen in bile duct cells and in hepatocytes. Levels of 27-kD heat-shock protein were also increased in some of the primary biliary cirrhosis biopsy specimens. The expression of progesterone receptor and androgen receptor was very low in normal and primary biliary cirrhosis bile duct cells and hepatocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225216 TI - Preoperative measurement of segmental liver volume of donors for living related liver transplantation. AB - Segmental liver volume determination by computed tomographic scan was carried out preoperatively in nine donors for living related liver transplantation. The calculated volume was compared with the graft size actually obtained by three types of donor hepatectomy. The volume of the left lateral segment (175 to 241 ml) and the left lobe (310 to 490 ml) varied markedly among the donors. The ratio of the left lobar to total liver volume also showed a wide range of values (23.2% to 35.9%). The value of the left lobar volume did not correlate positively with the donor's body weight, suggesting that graft size cannot be predicted only on the basis of the donor's body size. Segmental graft liver volume was estimated by use of computed tomographic scan, with acceptable accuracy on comparison with the graft volume actually obtained. In living related liver transplantation, the type of donor hepatectomy should be selected on the basis of the segmental liver volume of the donor in addition to the recipient's body size so that liver failure can be prevented in recipients and the donor's safety can be assured as far as possible. PMID- 8225217 TI - Treatment of small hepatocellular carcinoma in cirrhotic patients: a cohort study comparing surgical resection and percutaneous ethanol injection. AB - This study was intended to compare the survival rates of two contemporary cohorts of patients with solitary hepatocellular carcinomas < or = 4 cm subjected to surgical resection (n = 33) or percutaneous ethanol injection (n = 30). Outcomes in a third cohort, 21 patients with hepatocellular carcinoma who underwent orthotopic liver transplantation, were also assessed. Surgical and ethanol treated patients were similar with regard to age and tumor stage, differing only in liver function; 30 of the 33 surgical patients were of Child-Pugh class A, whereas only 7 of the 30 ethanol-treated patients were of class A (p < 0.05). Surgical resection was successful in 30 cases; ethanol injection achieved initial success in 23 patients. Tumor recurrence rate at 2 yr was 45% in the surgical group and 66% in the ethanol group. The difference was significant only for cases with tumors between 3 and 4 cm. Despite poorer liver function, the 1-, 2-, 3- and 4-yr survival rates of ethanol-treated patients (83%, 66%, 55% and 34%, respectively) were not different from those of surgically treated patients (81%, 73%, 44% and 44%, respectively). The 1- and 2-yr survival rates of patients given liver transplants were 81% and 66%, without tumor recurrence, after 16-mo follow up. These data confirm that ethanol injection is a useful treatment for patients with solitary small hepatocellular carcinomas and suggest that surgical resection and liver transplantation may achieve better results only after strict candidate selection to reduce mortality and tumor recurrence during follow-up. PMID- 8225218 TI - Primary sclerosing cholangitis and Hodgkin's disease. AB - Three patients with primary sclerosing cholangitis and Hodgkin's disease, a previously unrecognized association, are reported. All three patients were men, and one patient had Crohn's disease of the colon. Primary sclerosing cholangitis was diagnosed 2, 11 and 17 yr before diagnosis of Hodgkin's disease in the three patients, and all three had advanced biliary cirrhosis prompting referral for liver transplantation. The symptoms of Hodgkin's disease were often masked by similar manifestations of primary sclerosing cholangitis, particularly symptoms of recurrent biliary sepsis. Hodgkin's disease is another disorder that may occur in patients with primary sclerosing cholangitis, particularly in the setting of advanced disease, and may be masked by the underlying hepatobiliary disease. PMID- 8225219 TI - Circulating interleukin-1 and tumor necrosis factor antagonists in liver disease. AB - The proinflammatory cytokines interleukin-1 and tumor necrosis factor-alpha are thought to play important roles in the pathophysiology of liver disease. Specific antagonists of these cytokines have been found in recent years. Interleukin-1 receptor antagonist is a specific interleukin-1 antagonist. The soluble receptor derived from the cell-surface p55 tumor necrosis factor receptor p55 is a naturally occurring substance that inhibits the biological effects of tumor necrosis factor. We used specific radioimmunoassays to detect circulating interleukin-1 receptor antagonist and tumor necrosis factor soluble receptor p55 levels in 14 patients with acute viral hepatitis and in 160 patients with various chronic liver diseases. Levels of interleukin-1 receptor antagonist and, especially, tumor necrosis factor soluble receptor were markedly increased in most patients with chronic liver disease regardless of pathogenesis and in viral hepatitis. Patients with chronic liver disease and cirrhosis showed significantly higher levels of both cytokine antagonists than did noncirrhotic patients. Correlations between interleukin-1 receptor antagonist and tumor necrosis factor soluble receptor were more significant than those of either antagonist with C reactive protein or blood sedimentation rate. Interleukin-1 receptor antagonist and tumor necrosis factor soluble receptor levels were also positively correlated with bilirubin and AST levels. We conclude that circulating levels of interleukin 1 receptor antagonist and tumor necrosis factor soluble receptor may reflect ongoing disease activity and probably modulate some effects of endogenous interleukin-1 and tumor necrosis factor. PMID- 8225220 TI - Increased serum nitrite and nitrate levels in patients with cirrhosis: relationship to endotoxemia. AB - Nitric oxide derived from vascular endothelium is a potent vasodilator that plays a key role in the homeostasis of blood pressure. Because cirrhotic patients tend to have low arterial pressure, we measured in 51 patients and 10 control subjects serum nitrite and nitrate levels as an index of in vivo nitric oxide generation. We also measured plasma endotoxin, a substance frequently increased in cirrhotic patients and known to induce nitric oxide synthesis. Cirrhotic patients showed significant increases in serum nitrite/nitrate and plasma endotoxin compared with controls. Values were particularly increased in patients with decompensated cirrhosis, as manifested by ascites with or without functional kidney failure. High serum nitrite/nitrate levels were associated with high plasma renin activity, high aldosterone and antidiuretic hormone levels and low urinary excretion of sodium. In addition, serum nitrite/nitrate levels significantly correlated with endotoxemia. Oral administration of colistin to 15 cirrhotic patients reduced significantly plasma endotoxin levels (p < 0.01) and serum nitrite/nitrate levels (p < 0.05). Because endotoxin enhances the expression of inducible nitric oxide synthase, our results suggest that circulating endotoxin in cirrhosis is responsible for excessive synthesis and release of nitric oxide by the vasculature. These findings might explain the hemodynamic dysfunction seen in cirrhotic patients. PMID- 8225221 TI - Experimental evidence that the physiological position of the liver within the circulation is not a major determinant of zonation of gene expression. AB - Many enzymes are distributed heterogeneously within the liver lobule. The factors that play a determining role in the establishment and maintenance of these heterogeneous expression patterns have not yet been identified. To investigate whether the composition of the afferent hepatic blood plays a crucial role in the maintenance of the heterogeneity of gene expression of the parenchymal cells within the liver lobule, we changed the source of the afferent hepatic blood by microsurgical techniques. Three different groups of experimental animals were studied: rats with livers that are perfused with portal blood only (ligation of the hepatic artery), with caval blood only (portocaval transposition and ligation of the hepatic artery) and arterial blood only (portocaval shunt, arterialization of the distal end of the portal vein and ligation of the hepatic artery). To study differences in gene expression patterns, we chose enzymes that have a heterogeneous expression pattern within the liver lobule: the periportally located enzymes carbamoylphosphate synthase, succinate dehydrogenase, phosphoenolpyruvate carboxykinase and the pericentrally located enzymes glutamine synthase, glutamate dehydrogenase and NADPH-cytochrome P-450 reductase. To eliminate the potential interference of the long half-lives of some of these proteins on the interpretation of the results, we also studied the distribution of the mRNAs of carbamoylphosphate synthase, glutamine synthase, glutamate dehydrogenase and phosphoenolpyruvate carboxykinase. The animals were studied 2 wk after the operations. On the basis of their changes in body weight the animals were in steady state for at least a week. The patterns of gene expression of the enzymes studied did not change, regardless of the source of the altered afferent hepatic blood. The changes in gene expression that were observed in animals that did not regain their preoperative weight were shown to be caused by a limited intake of food. This study demonstrates that the physiological position of the liver within the circulation (i.e., between the gastrointestinal tract and the systemic circulation) is not as critical as is often stated and is certainly not essential for the maintenance of liver cell heterogeneity. The data suggest that the direction of the bloodstream (i.e., the existence of an upstream and a downstream compartment) is a major determinant of zonation of gene expression. PMID- 8225222 TI - Oxygen free radical production mediated by cocaine and its ethanol-derived metabolite, cocaethylene, in rat hepatocytes. AB - Cocaine ethyl ester ("cocaethylene") is a cocaine metabolite formed by hepatic carboxylesterases in the presence of ethanol. In the human liver, this pharmacologically active cocaine analog may reach concentrations similar to those of cocaine. To further investigate the intrinsic hepatotoxic potential of cocaethylene and to compare its effects with the known hepatocyte-damaging effects of cocaine, primary short-term cultures of hepatocytes derived from phenobarbital-pretreated rats were exposed to cocaine or cocaethylene (10 to 1000 mumol/L). Both drugs caused time- and concentration-dependent release of lactate dehydrogenase into the extracellular medium in a congruent pattern. Spontaneous hydrolysis of cocaethylene was negligible at pH 7.4, and the rate of 2 diethylaminoethyl-2,2-diphenylvalerate (SKF-525A) and phenylmethylsulfonylfluoride-sensitive enzymatic degradation in hepatocyte cultures was similar to that of cocaine (half-time, approximately 3 hr). Furthermore, both cocaine and cocaethylene induced massive NADPH-dependent reactive oxygen species production in hepatic microsomal suspensions and in homogenates of cultured hepatocytes, as directly demonstrated by superoxide dismutase- and catalase-sensitive luminol chemiluminescence. The integrated photon emission and the maximal slopes of the luminol chemiluminescence displayed similar concentration-response curves for cocaine and cocaethylene. These data suggest that reactive oxygen species, generated during the interaction of cocaine and cocaethylene with cytochrome P-450, may be the common mediators of hepatotoxicity induced by these compounds. Moreover, they indicate that the potential for cocaethylene to induce acute lethal cell injury in hepatocytes through oxidative mechanisms is similar to that of its parent compound, cocaine. PMID- 8225223 TI - Parallel decrease of Na(+)-taurocholate cotransport and its encoding mRNA in primary cultures of rat hepatocytes. AB - We investigated the molecular mechanism underlying the progressive loss of Na(+) dependent bile salt uptake in primary cultured rat hepatocytes. A specific cDNA probe was used to quantitate the levels of mRNA encoding the Na(+)-taurocholate cotransporting polypeptide at various culture times. Hepatocytes were cultured on collagen in the presence of insulin (10(-7) mol/L), dexamethasone (10(-7) mol/L) and 10% fetal calf serum for up to 72 hr. During this time period the dissociation constant of Na(+)-dependent taurocholate uptake remained stable (19 to 39 mumol/L), whereas the maximum velocity values decreased from 100% at 3 hr to 55%, 22% and 4% at 24, 48 and 72 hr, respectively. Concomitantly the levels of the Na(+)-taurocholate-cotransporting polypeptide mRNA also decreased from 100% at 3 hr to 41%, 24% and 4% at the later time points. In contrast, Northern hybridization with complementary DNA probes for three common housekeeping gene products revealed a 1.8- to 3.4-fold increase in the levels of mRNA encoding the alpha-subunit of the Na+K(+)-ATPase, beta-actin and glycerol-3-phosphate dehydrogenase. These data indicate that the loss of Na(+)-dependent bile salt uptake in primary cultures of rat hepatocytes is caused by decreased levels of its specific mRNA. Hence the studies further confirm that without specific measures (primary) cultured rat hepatocytes reverse their liver-specific phenotype to a more fetal pattern of gene expression. PMID- 8225224 TI - Vasoactive effects of bile salts in cirrhotic rats: in vivo and in vitro studies. AB - To clarify a possible pathogenic role for bile salts in the hyperdynamic circulation of cirrhosis, we studied the vasoactive effects of three different bile salts-tauroursodeoxycholic acid, taurochenodeoxycholic acid and taurodeoxycholic acid-in cirrhotic rats. Cirrhosis was induced with bile duct ligation; controls underwent sham surgery. In vivo, the bile salts were intravenously infused at one of three doses (1.2 x 10(-7), 1.2 x 10(-6) and 6.0 x 10(-5) mol x 100 gm-1 x min-1) for 5 min. Taurochenodeoxycholic acid and taurodeoxycholic acid infusions increased mesenteric arterial blood flow and conductance and induced systemic arterial hypotension, whereas tauroursodeoxycholic acid had no significant effect. At similar plasma levels of bile salts, the responses in cirrhotic rats were attenuated compared with those of controls. In vitro, isolated rings of superior mesenteric and carotid arteries and portal vein were precontracted with phenylephrine; then dilatory responses to cumulative doses of bile salts (10(-6) to 10(-3) mol/L) were measured. In all three vessels, taurodeoxycholic acid produced stronger dilatory effects than did taurochenodeoxycholic acid, whereas tauroursodeoxycholic acid showed no significant effect. Vessels from cirrhotic and control rats did not differ in degree of response. These results indicate that bile salts are directly vasoactive and can induce splanchnic vasodilation at the pathophysiological plasma levels seen in cirrhosis. Bile salts may be involved in the pathogenesis of splanchnic hyperemia and hyperdynamic circulation in cirrhosis. PMID- 8225225 TI - Bile acid-induced modifications in DNA synthesis by the regenerating perfused rat liver. AB - Liver cell proliferation is a complex process that can be affected by a large number of factors such as bile acids, which have been reported to be associated to the pathogenesis of liver cancer. In this work, bile acid-induced modifications in DNA synthesis by regenerating perfused rat liver were investigated. Two-thirds hepatectomy was carried out 24 hr before perfusion of liver with recirculating, erythrocyte-free Krebs-Henseleit solution. The viability of the preparations was maintained under all experimental conditions, as indicated by bile flow, oxygen uptake, perfusion pressure, perfusion flow and release of lactate dehydrogenase and potassium into the perfusate. Livers received (min 10 to min 60) bile acid infusion at a rate of 25 nmol/min/gm liver (i.e., maximal secretion rate/2) in regenerating livers as calculated for taurocholate in separate experiments). Trace amounts of [methyl-14C]thymidine were added to the perfusate at min 30. At the end of the experiments (min 60) the livers were washed, removed, weighed and homogenized to determine radioactivity in whole tissue, in DNA and in non-DNA-related fractions. Taurocholate and, to a lesser extent, taurodeoxycholate and dehydrocholate (but not ursodeoxycholate) were found to reduce 14C incorporation into DNA. This was not due to changes in the content of 14C in whole, regenerating liver tissue. Taurocholate, taurodeoxycholate, dehydrocholate and ursodeoxycholate had no effect on thymidine uptake; moreover, the proportion of 14C found in bile was negligible. However, bile acid-induced modification in the fate of intracellular thymidine was observed. In regenerating livers receiving no bile acid, the 14C carried by thymidine metabolites accounted for about 60% of 14C in whole liver tissue. Taurocholate markedly increased this proportion to about 80%. Reverse-phase high pressure liquid chromatography revealed that most of this 14C (about 80%) was recovered at the elution time, corresponding to thymidine catabolites rather than to DNA precursors. These results suggest that bile acids induce enhancement of thymidine catabolism that reduces its incorporation into DNA; inhibition in the process of DNA synthesis itself, leading to a subsequent increase in the metabolism of DNA precursors; or both. Moreover, from the diversity in this property for bile acid species it might be inferred that changes in the composition and size of the bile acid pool during liver carcinogenesis or regeneration play a role in the modulation of the proliferative process. PMID- 8225226 TI - Induction of metallothionein and its localization in the nucleus of rat hepatocytes after partial hepatectomy. AB - Metallothioneins, a group of cysteine-rich heavy-metal binding proteins, are induced in the regenerating rat liver in response to the stimuli evoked by partial hepatectomy. We have investigated the expression of metallothionein genes and proto-oncogenes (c-fos, c-jun and c-myc), as well as specific localization of metallothionein in the liver cells after partial hepatectomy. Metallothionein mRNA was detected as early as 3 hr and reached a maximal level by 6 hr. Expression of the proto-oncogenes apparently preceded the elevation of metallothionein protein because the latter was maximal 18 hr after partial hepatectomy, followed by a decrease until 70 hr. Hepatocytes of the intact rat liver have metallothionein in the cytoplasm only. Interestingly, metallothionein was localized predominantly in the nucleus as early as 6 hr after partial hepatectomy, and the staining intensity of metallothionein became maximal at 15 hr, followed by detection in both the cytoplasm and nucleus at 24 hr or longer. The use of a confocal laser scanning microscope with both tissue sections and isolated nuclei has clearly shown that metallothionein immunofluorescence exists inside hepatocyte nuclei after partial hepatectomy. Expression of the proto oncogenes c-fos and c-jun is elevated after partial hepatectomy, and the resultant heterodimer of gene products may contribute to the observed metallothionein gene induction. However, the observation that metallothionein protein levels were elevated until 18 hr after partial hepatectomy suggests that an alternative pathway for the induction of metallothionein gene expression may also be present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225227 TI - Modulators of the protein kinase C system influence biliary excretion of cationic drugs. AB - To investigate whether hepatobiliary transport of organic cations is under regulatory control, we studied transport of tri-n-butylmethylammonium in the isolated perfused rat liver and in isolated rat hepatocytes. Transport was investigated in the presence of modulators of the protein kinase C and the cyclic AMP second-messenger system. In the isolated perfused rat liver, it was observed that compounds modulating protein kinase C activity clearly affected the biliary excretion process of the cation tri-n-butylmethylammonium. Phorbol 12-myristate 13-acetate, a compound that directly stimulates protein kinase C, elevated the biliary excretion rate of tri-n-butylmethylammonium in a concentration-dependent manner, reaching a twofold increase at 60 nmol/L of the phorbol ester. The inactive derivative 4 alpha-phorbol 12, 13-didecanoate (60 nmol/L) did not show any effect. Vasopressin (48 nmol/L), a receptor-mediated activator of protein kinase C, stimulated the excretion rate of the cation by about 50%. Staurosporin (1 mumol/L), an inhibitor of protein kinase C, clearly decreased the biliary excretion rate of the cation and also blocked its stimulation by phorbol 12 myristate 13-acetate. Neither phorbol 12-myristate 13-acetate nor vasopressin (at concentrations ranging from 10(-9) to 10(-6) mol/L) affected the initial uptake velocity of tri-n-butylmethylammonium in isolated hepatocytes and isolated perfused livers, whereas staurosporin (1 mumol/L) showed only a modest inhibition of the uptake of the cation. It is inferred that the effect of protein kinase C modulators on hepatobiliary transport of organic cations occurs at the level of carrier-mediated transport in the canalicular membrane.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225228 TI - Membrane-associated C-reactive protein on rat liver macrophages is synthesized within the macrophages, expressed as neo-C-reactive protein and bound through a C reactive protein-specific membrane receptor. AB - We could recently show that rat liver macrophages (Kupffer cells) express a membrane-bound form of C-reactive protein on their surface. Because it is removed by washing the cells in buffers containing Ca(++)-chelators, membrane-bound C reactive protein is a peripheral protein rather than an integral part of the Kupffer cell membrane. This Kupffer cell membrane-bound C-reactive protein is identical to the galactose-specific particle receptor previously characterized. We now present evidence that Kupffer cells do not acquire soluble serum C reactive protein but synthesize their own membrane-bound C-reactive protein. By RNA-RNA in situ hybridization, it was found that hepatocytes are not the only sort of liver cells synthesizing C-reactive protein, but C-reactive protein specific mRNA is present also in Kupffer cells. During acute-phase response C reactive protein mRNA is found in increased amounts within liver macrophages too. Furthermore, by labeling experiments with antisera against native, pentameric soluble serum C-reactive protein and monoclonal antibodies against a neoepitope present on C-reactive protein subunits only, we could establish that the membrane bound C-reactive protein expressed on the liver macrophage is not the pentameric molecule of soluble serum C-reactive protein, but rather consists of C-reactive protein subunits. Finally, we present evidence that liver macrophages contain a binding protein in their plasma membrane, with an apparent molecular weight of 59 to 61kD, specific for C-reactive protein and similar to the one previously isolated from macrophage cell lines. PMID- 8225229 TI - Selectivity and sensitivity of changes in serum bile acids during induction of cirrhosis in rats. AB - Because some patients with cirrhosis have serum transaminase levels within the normal range, a prospective study was undertaken to determine whether the concentration of individual serum bile acids would be a sensitive indicator of development of cirrhosis. The choline-deficient rat has been used as a model for study of these changes. Using high-performance liquid chromatography, we measured the concentrations of individual serum bile acids at 3, 6, 10, 20 and 30 wk of dietary intake. Serum levels of total glycine- and taurine-conjugated bile acids were elevated at all stages tested as compared with levels in control groups (choline supplemented). Similarly, unconjugated bile acids and, particularly, cholic acid showed significantly higher levels at all stages except with the occurrence of cirrhosis at 30 wk, at which time there was a significantly lower level for unconjugated bile acids (0.48 +/- 0.11 vs. 1.40 +/- 0.36 in controls) and for cholic acid (0.17 +/- 0.05 vs. 0.91 +/- 0.39 in controls). The ratio of serum cholic acid to serum chenodeoxycholic acid changed in temporal relationship to progression in the histological lesions in livers of these rats. The ratio was at its highest at 78 +/- 3 at 3 wk (no histological change) and decreased with increasing time and changes in histological appearance until 30 wk, at which time it was down to 1.6 +/- 0.6. The routinely used markers of liver injury (serum ALT, alkaline phosphatase and bilirubin), however, did not match the progression of hepatic histological changes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225230 TI - Inhibitory effect of gastrin and cholecystokinin fragments on the secretion of triacylglycerol in rat hepatocytes. AB - In this investigation we studied the influence of two gastrin fragments, pentagastrin and nonsulfated heptadecagastrin, and two cholecystokinin fragments, sulfated and desulfated cholecystokinin 26-33, on intracellular and secreted triacylglycerol in isolated hepatocyte cultures. Both gastrin fragments inhibited triacylglycerol release in a biphasic manner, exhibiting maximal effect at 0.1 nmol/L (nonsulfated heptadecagastrin) and 0.3 nmol/L (pentagastrin). At these concentrations triacylglycerol secretion was 42% (nonsulfated heptadecagastrin, p < 0.001) and 62% (pentagastrin, p < 0.001) lower than in cells untreated with gastrin. Sulfated cholecystokinin 26-33 caused a 35% decrease in triacylglycerol secretion at 0.1 nmol/L (p < 0.01), and desulfated cholecystokinin 26-33 caused a 53% decrease at 0.2 nmol/L (p < 0.001). In all experiments, the hormone-induced decrease in triacylglycerol secretion was accompanied by an increase in intracellular triacylglycerol content. The cholecystokinin-A receptor antagonist L-364, 718 did not affect the decrease in triacylglycerol secretion induced by 0.3 nmol/L pentagastrin, whereas the cholecystokinin-B receptor antagonist L-365, 260 inhibited the pentagastrin effect at concentrations above 50 nmol/L. These results suggest that gastrin, cholecystokinin or some other gastrinlike hormone (or all three) may play a previously unrecognized regulatory role with respect to hepatic very low density lipoprotein secretion. PMID- 8225231 TI - Apoptosis is induced by transforming growth factor-beta 1 within 5 hours in regressing liver without significant fragmentation of the DNA. AB - In previous studies we showed that transforming growth factor-beta 1 induces apoptosis in hepatocyte cultures and regressing livers, the mature form being more potent than the transforming growth factor-beta 1 latency-associated protein. In this study we addressed the question of whether apoptosis can be induced within a short time after administration of transforming growth factor beta 1. Five hours after a single intravenous injection of 25 micrograms mature transforming growth factor-beta 1/kg body weight, apoptosis is augmented ninefold in the regressing rat liver. A second preceding application induces no further augmentation. Transforming growth factor-beta 1 latency-associated protein shows no effect with either regimen. Morphological evaluation shows that 5 hr after injection of transforming growth factor-beta 1 nearly all apoptotic bodies are already engulfed by their neighbor cells. After homogenization of the transforming growth factor-beta 1-treated livers, the condensed apoptotic bodies are not destroyed and remain in the nuclear pellet. No DNA fragmentation into oligosomes could be detected after purification of the DNA from the nuclear pellet and application to conventional gel electrophoresis. Application of in situ nick translation, which allows detection of DNA single- and double-strand breaks in individual apoptotic bodies, also revealed no substantial fragmentation of the DNA in apoptotic bodies. These studies show that transforming growth factor-beta 1 is able to induce apoptosis within a rather short time and also suggest that in vivo digestion of the DNA does not lead to chromatin condensation. PMID- 8225232 TI - Alcohol consumption enhances fatty acid omega-oxidation, with a greater increase in male than in female rats. AB - Because ethanol inhibits mitochondrial fatty acid oxidation, with substantial accumulation of fatty acids in the livers of female (but not male) rats, and induces microsomal activities, we assessed possible changes in omega-oxidation. To study this, we pair-fed 24 male and 24 female littermate rats of the same age liquid diets containing 36% of energy either as ethanol or as additional carbohydrate for 4 wk. In controls, the microsomal omega-hydroxylation of lauric acid was 28% greater in female than in male rats (p < 0.05). Ethanol feeding significantly increased this activity in both genders (p < 0.01), but the rise in male rats (89%) was significantly higher than that in female rats (24%). This activity was unaffected by the presence of ethanol in the assay. The effects of ethanol were associated with increases in the content of cytochrome P-450 4A1 (as assessed in Western blots by the reactivity against a sheep antibody against P 450 4A1), and more so in male than in female rats. Despite possible competition by ethanol with the hydroxy fatty acid oxidation to dicarboxylic acids through alcohol dehydrogenase, suberic and sebacic acids accumulated significantly in the livers of alcohol-fed male rats. These effects of ethanol and gender on omega oxidation paralleled those on the hepatic cytosolic fatty acid-binding protein and fatty acid esterification previously reported in similarly treated rats. Dicarboxylic acid products of omega-oxidation have been incriminated as mediators of similar effects by other drugs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225233 TI - Regulation of human liver cytochromes P-450 in family 3A in primary and continuous culture of human hepatocytes. AB - The cytochrome P-450 3A gene family comprises the dominant forms of cytochrome P 450 found in human liver. We examined as a possible useful system for studying the regulation of cytochrome P-450 3A under controlled conditions in vitro, primary monolayer cultures of human hepatocytes and compared the results with those obtained from the study of cytochrome P-450 3A in the human hepatoblastoma cell line HepG2 or in the human hepatocellular carcinoma cell line TONG/HCC. Using 3A antibodies, 3A cDNAs and 3A3, 3A4, 3A5 and 3A7 isozyme-specific oligonucleotides as probes, we determined that primary human hepatocyte cultures routinely expressed a 3A3/4* immunoreactive protein and 3A mRNA. These gene products were well maintained for many days and were induced by treatment of the cultures with dexamethasone, phenobarbital, macrolide antibiotics, the HMG CoA reductase inhibitor lovastatin or an antifungal agent, clotrimazole. Of six donor livers examined, only two contained mRNA or protein for 3A5, a form found in only a few adult human subjects. In cultures prepared from one of these two livers, 3A5 mRNA was detectable for several days. In cultures of hepatocytes from the remaining four human livers that did not contain 3A5 mRNA or protein, we detected neither spontaneous nor inducible 3A5 proteins or mRNAs. HepG2 cells contained only 3A7 protein, a form found in human fetal liver, even after treatment with inducers. treatment of HepG2 cells with dexamethasone, macrolide antibiotics, phenobarbital and phenobarbital-like inducers or lovastatin produced dose dependent induction of 3A7 mRNA and 3A7 immunoreactive protein. TONG/HCC cells contained 3A3, 3A4 and 3A5 mRNAs, but only 3A5 immunoreactive protein could be detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225234 TI - Hepatic gene therapy: present and future. PMID- 8225235 TI - Bile acids for babies? Diagnosis and treatment of a new category of metabolic liver disease. PMID- 8225236 TI - A new bile acid transporter? PMID- 8225237 TI - True transport: one or more sodium-dependent bile acid transporters? PMID- 8225238 TI - Soft tissue tumours in childhood. AB - The spectrum of soft tissue tumours in young adults is very similar to that in more mature individuals, while those in childhood form a distinct group rarely seen outside the first decade. The majority of these are benign vascular or fibroblastic proliferations; in young children they may be highly cellular and mitotically active, but malignancy should be diagnosed with caution. Congenital soft tissue tumours constitute a special group in which the clinical outcome may be particularly difficult to predict from the histological appearances. This review focuses on those malignant soft tumours which are either peculiar to childhood or which manifest special features in childhood. Some recently recognized benign soft tissue tumours which occur mainly in childhood are also described. The fibromatoses are not discussed. As a guide to the appropriate treatment regime, the main histological distinctions to be drawn are between: 1 tumours of neuroectodermal origin (Ewing's sarcoma and primitive neuroectodermal tumour); 2 other sarcomas; and 3 the fibromatoses and other benign and potentially locally aggressive lesions requiring local excision. Immunohistochemical staining may be of considerable help in achieving the correct diagnosis, but it is vital that a panel of antibodies be applied and the results critically assessed. Cytogenetic analysis is also of growing importance, characteristic karyotypic abnormalities having been demonstrated in Ewing's sarcoma/primitive neuroectodermal tumour, alveolar rhabdomyosarcoma and synovial sarcoma. PMID- 8225239 TI - T-cell-rich B-cell lymphoma presenting as liver disease. AB - We describe a series of eight cases of T-cell-rich B-cell lymphoma diagnosed on liver biopsy and collected over a period of 15 years. Of seven cases that were referred from elsewhere, in only one was the correct diagnosis of B-cell lymphoma suggested. Common errors included misdiagnosis as inflammatory disease on histology, and misinterpretation as T-cell lymphoma on immunohistochemistry. However, the cases had a distinct morphological appearance and immunohistochemical profile. They showed a lymphohistiocytic or granulomatous infiltrate, usually centred on portal tracts and containing abundant small T cells and scanty B-cell blasts. All patients had an atypical clinical presentation which favoured non-neoplastic liver disease. In seven cases liver involvement represented Stage IV disease and in one case disease was confined to the liver consistent with a primary hepatic lymphoma. Despite combination chemotherapy, the prognosis was poor with no patients surviving beyond 15 months from diagnosis. We believe T-cell-rich B-cell lymphoma to be an under-recognized subset of non-Hodgkin's lymphoma that may mimic primary liver disease. PMID- 8225240 TI - Carcinoma and atypical hyperplasia in radial scars and complex sclerosing lesions: importance of lesion size and patient age. AB - One hundred and twenty-six radial scars and complex sclerosing lesions from 91 women were examined to determine the incidence of and the clinical and pathological factors associated with the development of carcinoma and atypical hyperplasia within them. There was a clear relationship between the presence of carcinoma and atypical hyperplasia and the size of the lesion. This was not, however, a progressive relationship, there being a cut-off point about 6-7 mm, below which carcinoma was very uncommon and above which it was relatively frequent. A similar relationship was seen with patient age. Carcinoma was not seen in lesions removed from women under 40, was rare in the decade 41-50 and was relatively common above this age but with no further increase in the over 60s. A significantly higher incidence of carcinoma and atypical hyperplasia was encountered in scars detected by mammographic screening and could be explained by lesion size and the ages of the patients from which they were removed. No relationship was found between the presence of carcinoma within radial scars and complex sclerosing lesions and the existence of carcinoma in the residual breast tissue when direct extension was excluded. The carcinomas identified in the scars were of variable type and included small and large cell ductal carcinoma in situ, lobular carcinoma in situ and invasive carcinoma of tubular and ductal types. In situ carcinoma and atypical hyperplasia involved a very variable percentage of the epithelium of the lesions with mean values for ductal carcinoma in situ of 32%, lobular carcinoma in situ 25% and atypical hyperplasia 25%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225241 TI - Methodology of immunohistological detection of oestrogen receptor in human breast carcinoma in formalin-fixed, paraffin-embedded tissue: a comparison with frozen section methodology. AB - We describe a method of immunohistochemically assessing estrogen receptor status on routinely processed formalin-fixed tissue, using a commercially available monoclonal antibody (Abbott H222), with pronase predigestion of tissue sections and overnight antibody incubation. The staining was assessed using the H score system. A series of 94 cases of breast cancer were analysed and the results were compared with assessment by oestrogen receptor immunocytochemical assay performed on frozen section. Direct comparison of the paired sets of H scores obtained with frozen tissue and formalin-fixed tissue showed a highly significant correlation of 0.8 (P < 0.001) between the two methods of oestrogen receptor assessment. Chi squared analysis using H score cut off points of 50 and 100 also showed a similar significant association (P < 0.001). We conclude that this oestrogen receptor method, applicable to formalin-fixed, paraffin-embedded tissue, gives accurate results on routinely fixed tissue and could be used as an alternative to other methods. PMID- 8225242 TI - Oestrogen receptor, progesterone receptor, pS2, ERD5, HSP27 and cathepsin D in invasive ductal breast carcinomas. AB - Hormonal receptors and markers for prognostic evaluation were detected immunohistochemically in 196 infiltrating ductal breast carcinomas. Immunohistochemical detection of progesterone and oestrogen receptor is a method giving results generally concordant with those of the binding assay. However, immunohistochemical detection seems better. It allows the detection of hormonal receptors on small carcinomas, it is not modified by the endogenous hormones, and it has a slightly better correlation with prognosis and with the response to hormone therapy. Immunohistochemical detection of progesterone receptor has a prognostic value, sorting a negative subgroup with a poor prognosis from the oestrogen receptor positive tumours. These results can be obtained without quantitative immunohistological methods. ERD5, pS2, HSP27 and cathepsin D are associated with oestrogen receptor positivity. pS2 and HSP27 are interesting markers. They characterize a subgroup of oestrogen receptor negative tumours with a good prognosis. Moreover, pS2 is a marker of response to hormone therapy. ERD5 and cathepsin D do not appear to be of value as markers of prognosis. PMID- 8225243 TI - Immunohistochemistry of pS2 in normal human breast and in various histological forms of breast tumours. AB - Expression of pS2 was studied by immunocytochemistry in normal breast tissue (n = 20), benign tumours (n = 9) and 145 breast cancers representative of the different histological types. pS2 immunostaining was scored as negative (D1 = 0 5% stained cells), positive (D2 = 5-75% stained cells) or highly positive (D3 > 75% stained cells). pS2 protein was evident in all normal breast samples examined. Six of nine benign lesions showed pS2 staining. In both cases, immunostaining was weaker than in breast cancers. Of breast cancers, 77/145 (53.1%) were pS2 positive, including 33.1% with intense staining. The presence of pS2 was not correlated with the age of patients, the size of the primary tumour, or lymph node status, but was correlated with histological grading and nuclear grading. pS2 expression was also correlated with menopausal status and oestrogen receptor status (59% of receptor-positive tumours were pS2 positive), but not to progesterone receptor status. pS2 expression in breast carcinomas is not a characteristic of specific histological types. Although this protein is predominantly expressed in oestrogen receptor-positive and differentiated tumours, it shows oestrogen-independent expression in about 30% of cases. PMID- 8225244 TI - Immunohistochemically demonstrated metallothionein expression in malignant melanoma. AB - Metallothioneins are ubiquitous proteins with a high affinity for heavy metal ions, e.g. zinc, copper and cadmium. Experimentally, metallothionein over expression in cell lines derived from a variety of cancers has been associated with resistance to anticancer drugs and irradiation therapy. Using a monoclonal antibody (E9) to metallothionein we investigated immunoreactive expression in routinely fixed and paraffin-embedded tissue from 63 cases of malignant melanoma and 13 secondary deposits. Whereas a variety of cells in normal skin showed metallothionein expression, all forms of benign naevi studied were uniformly negative. In contrast 13/30 'thin' (< or = 1.5 mm; 0.7 +/- 0.4), 25/29 'thick' malignant melanoma (> 1.5 mm; 5.5 +/- 3.9) and 12/13 metastases were positive. Six patients with thin and 19 with thick melanoma with metallothionein expression died during a mean observation period of 6.4 +/- 1.8 and 3.6 +/- 2.5 years, respectively, their survival distribution function analyses giving statistically significant results for both the vertical tumour thickness (P < 0.0001) and metallothionein expression (P < 0.0001). These immunohistochemical results, based on routinely processed paraffin-embedded tissue, suggest that metallothionein expression in malignant melanoma is significantly associated with progressive disease and might therefore be a useful prognostic indicator. PMID- 8225245 TI - A histological morphometric study of nuclear size in benign and malignant neoplasms of the human cheek. AB - This report describes the application of simple morphometric methods to generate quantitative data on nuclear size from tissue sections of normal, benign and malignant oral epithelium of the cheek. Measurements of nuclear areas of basal and spinous cells from cheek mucosa were made on haematoxylin and eosin-stained sections using a Zeiss VIDAS image analyser. The lesions examined comprised benign squamous cell papillomas and islands of cells from invasive squamous cell carcinomas. Normal control epithelium was obtained from the biopsy margins of non neoplastic lesions. The nuclear areas (AN) were obtained by direct measurement whereas the nuclear diameters (DN) were determined automatically. In both basal and spinous strata, values for both these nuclear parameters were lowest in normal tissue and increased progressively through benign papillomas, with the highest values being found invariably in carcinomas. Statistically significant differences were detected between both normal and carcinoma and between papilloma and carcinoma. The morphometric parameters AN and DN are of value in distinguishing benign from malignant lesions of the human cheek. PMID- 8225246 TI - Enteric differentiation in cervical adenocarcinomas and its prognostic significance. PMID- 8225247 TI - Cerebral metastasis from a malignant mixed mullerian tumour of the uterus. PMID- 8225248 TI - Juxtacortical malignant schwannoma with heterologous elements. AB - A case of malignant schwannoma with heterologous elements occurring in the periosteum of a long bone is reported. We believe that this is the first time such a tumour has been reported at this site. PMID- 8225249 TI - Salivary-type myxoid myoepithelioma of the sinonasal tract: a potential diagnostic pitfall. AB - A 38-year-old woman presented with otalgia and facial pain. She was found to have a tumour involving the right maxillary sinus and pterygopalatine fossa. Exclusively composed of a myxoid population of spindle and stellate cells within a mucosubstance-rich matrix, it was originally interpreted as a mesenchymal neoplasm. However, immunophenotypic characterization revealed a pure population of salivary-type myoepithelial cells with cytoplasmic co-expression of cytokeratin, vimentin, muscle-specific actin, S-100 protein and glial fibrillary acidic protein. Sinonasal myxoid myoepithelial neoplasia is exceedingly rare and represents a potential diagnostic pitfall. PMID- 8225250 TI - Mixed medullary and follicular carcinoma of the thyroid. PMID- 8225251 TI - Cribriform intra-tubular epididymal change and testicular atrophy. PMID- 8225252 TI - Tubo-endometrioid metaplasia of the cervix may be post-traumatic. PMID- 8225253 TI - Reproductive behavior, endocrine state, and the distribution of GnRH-like immunoreactive mast cells in dove brain. AB - Previous studies indicate that there is an increase in the number of detectable mast cells expressing gonadotropin-releasing hormone-like immunoreactivity (GnRH ir) in the medial habenular region of the brain in ring doves after a period of 2 hr of courtship. In the present experiment the overall distribution of GnRH-ir mast cells in the brain is described, and the influence of behavioral and endocrine state on brain mast cell distribution and detectable mast cell number is explored. Four groups of male birds were studied: (1) paired with a female and displaying courtship behavior, (2) paired with a squab and displaying aggressive behavior, (3) visually isolated from other doves, and (4) long-term castrates housed communally. In all four groups, GnRH-ir mast cells were observed in the following areas: the medial habenula, circumventricular organs, organum vasculosum lamina terminalis and organum subseptale, the pia mater, and blood vessels. Cell counting revealed differences among groups in numbers of habenular mast cells. Courted doves had the most habenular mast cells, followed by doves paired with squabs, and then visually isolated doves, while castrates had the fewest. One-way ANOVA indicates significant differences among groups. All pairwise comparisons show that the courting animals had significantly more detectable mast cells in the habenular area than did visually isolated and castrated groups. These results suggest that the appearance and/or detectability of GnRH-ir mast cells in the habenula is related to the behavioral state and possibly to the endocrine state of the animal and suggest a novel mechanism for interactions among the nervous, endocrine, and immune systems. PMID- 8225254 TI - Hypothalamic and hypophyseal receptors for estradiol in high and low sexually performing rams. AB - Rams, characterized over a 3-year period as being high (n = 8) or low (n = 10) sexual performers, were euthanized to determine if differences in sexual behavior were associated with differences in hypophyseal and hypothalamic receptors for estradiol. Rams were exposed to estrous ewes and courtship behavior was recorded for 5 hr prior to tissue collection. Blood samples for determination of serum concentrations of luteinizing hormone (LH) were collected every 15 min during the behavior testing period. Hourly blood samples were analyzed for serum concentrations of estradiol (E) and testosterone (T). Rams characterized as high (HP) or low (LP) sexual performers had similar LH profiles; there were no differences noted in basal concentration, number of pulses, or pulse amplitude of LH. There were no differences in serum concentrations of T or E during the behavior testing period between HP and LP rams. Unoccupied receptors for E (moles E bound x 10(-15)) in the stalk-median eminence (SME) (P < 0.05) in HP rams (15.1 +/- 3.1) were greater than those in LP rams (4.6 +/- 1.9). Occupied receptors for E (moles E bound x 10(-15)) in the preoptic area (POA) (P < 0.1) in HP rams (6.0 +/- 0.8) tended to be greater than those in the LP rams (3.8 +/- 0.8). The proportion of total E receptors (occupied+unoccupied) in the POA that were occupied was greater (P < 0.05; 0.41 +/- 0.03 vs 0.23 +/- 0.04) for HP than for LP rams. In contrast, HP rams had less (P < 0.05) occupied E receptors (moles E bound x 10(-15)) in the anterior pituitary gland (AP) than LP rams (152.0 +/- 38.5 vs 337.3 +/- 51.7). The proportion (%) of E receptors that were occupied in the AP in HP rams also tended to be less (P < 0.06) than that in LP rams (8 +/- 2 vs 14 +/- 1). No differences were detected in occupied or unoccupied E receptors in the medial basal hypothalamus, medial amygdala, or stalk-median eminence. In conclusion, differences in occupied receptors for E in the POA and the AP, and unoccupied receptors for E in the SME, were associated with differences in ram sexual behavior. PMID- 8225255 TI - Mate choice and reproductive success in the red-legged partridge (Alectoris rufa). AB - Behavioral patterns, reproductive success, and concentrations of plasma luteinizing hormone, testosterone, and estrogen were compared in pairs of red legged partridges which had either selected their own mates or had been randomly paired. Males in self-selected pairs called more frequently than males in non self-selected pairs. Females in self-selected pairs but not those in non-self selected pairs guarded their nests. Females in self-selected pairs had a greater egg production and laid more of their eggs in nests than females in non-self selected pairs. There was a significant difference between the two groups in the concentration of plasma testosterone in males, which was higher in males from self-selected pairs prior to the onset of egg laying. These results are consistent with the hypothesis that mate choice affects both reproductive parameters and nest defense in monogamous birds. PMID- 8225256 TI - Relationship between social factors and pituitary-adrenocortical activity in female rhesus monkeys (Macaca mulatta). AB - Psychosocial stress in humans has been related to the occurrence or progression of certain diseases and a positive social environment has been shown, in some cases, to ameliorate this effect. In many experimental studies changes in serum cortisol levels have been used as an endocrine-defined stress response, however, the variation in this measure under unchanging conditions in socially housed animals is not well documented. The present study was designed to examine the relationship between specific social behaviors and concentrations of serum cortisol in female rhesus monkeys. Subjects were members of one of two social groups. One was an established long-term (8 years) group with members which had from one to many kin, while the second was a group formed 5 months prior to the onset of the present study composed of initially unfamiliar animals. Blood samples were collected weekly and a total of 16 hr of behavioral data was collected on each subject. For purposes of analyses, the year-long study was organized according to calendar quarters (3 months) with cortisol concentrations averaged and behavioral categories cumulated. When regression techniques were applied, it is of note that neither group nor dominance rank was a predictor of cortisol levels. However, when a correlation analysis was applied to each group separately to assess the effect of dominance rank, rank significantly correlated with cortisol levels in the established but not in the recently formed group. In each quarter, analyses revealed behavioral categories which combined to account for a significant proportion of the variance in cortisol levels, but these behaviors were not identical over the four quarters. For three of the four quarters analyzed, and for the year as a whole, an affiliative behavioral category was predictive of cortisol levels in addition to agonistic behavioral categories. These data suggest that cortisol levels are influenced by not only negative interactions, such as receiving bites, but also by positive interactions such as receiving grooms. Results also showed that reconciliatory behaviors occurred significantly more frequently following dyadic agonistic episodes in the recently formed versus the long-term group. The higher rate of reconciliatory behaviors exhibited by the recently formed group may account for the lack of a dominance rank/cortisol relationship. PMID- 8225257 TI - Sex steroid levels in Porichthys notatus, a fish with alternative reproductive tactics, and a review of the hormonal bases for male dimorphism among teleost fishes. AB - Plasma levels of 11-ketotestosterone, 11-beta-OH-testosterone, testosterone, and 17-beta-estradiol were measured in reproductive Porichthys notatus, a teleost fish with two male morphs and alternative reproductive tactics. The two male types had contrasting androgen profiles. 11-Ketotestosterone was the predominant androgen in the Type I male morph which acoustically courts females, excavates nests, and guards eggs. Yet testosterone was predominant in the plasma of the Type II male morph which neither courts females nor nests, but instead parasitizes Type I males with sneak or satellite spawning tactics. The Type I Type II male divergence in reproductive tactics and androgen levels is paralleled by dimorphisms in the vocal system, in body size, and in relative testis size. A review of endocrine data from six different species with male dimorphism shows consistent differences between morphotypes and a striking pattern: 11 ketotestosterone levels are uniformly elevated in each "courting male" morphotype relative to its "noncourting" conspecific. This cross-species pattern may reflect the behavioral, gonadal, or morphological differences which characterize the two morphotypes. At this point, the morphological interpretation is favored. PMID- 8225258 TI - Etiology of the sexual dimorphism in renal peripheral benzodiazepine receptor response to stress in rats. AB - A sexual dimorphism in stress-induced alterations in renal peripheral benzodiazepine receptors (PBR) was recently reported. The present paper includes five experiments examining the etiology of this sex difference. Surgical removal of ovaries and testes was ineffective in altering the renal PBR stress response in both male and female rats. A diurnal variation in the sexual dimorphism was observed; the difference was seen in the early part of the light cycle, while the two sexes were indistinguishable at the end of the light cycle. Finally, based on recent data indicating the importance of the renin-angiotensin system in the stress-induced decrease in renal PBR, we examined both stress-induced plasma renin activity and renal PBR reactivity to exogenous angiotensin II (AII) administration in both sexes. Female rats show both an attenuated elevation of plasma renin levels in response to inescapable shock stress and a reduced PBR response to AII administration in comparison to males. The present data indicate that the renin-angiotensin system may be a critical factor in the sexual dimorphism in the renal PBR response to stress. The possibility of this difference in the renin-angiotensin system contributing to sex differences in susceptibility to escape deficits following inescapable shock is entertained. The implications for these findings regarding the physiological function of the PBR are also discussed. PMID- 8225259 TI - Sex differences and the role of aromatization in the control of sexually dimorphic behavior and morphology in gray short-tailed opossums (Monodelphis domestica). AB - Sex differences in the activating effects of an aromatizable (testosterone, T) and a nonaromatizable (dihydrotestosterone, DHT) androgen on sexually dimorphic scent marking and aggressive behavior were examined in gonadectomized gray short tailed opossums. When compared with males, females showed less chest, head, flank, and hip marking and more fighting behavior in tests with stimulus females and threat behavior in tests with stimulus males following receipt of subcutaneous T, DHT, or blank (B) implants. Testosterone but not DHT activated hip and head marking while both T and DHT activated flank marking. In tests with stimulus females, only males showed clicking vocalizations and T but not DHT or B stimulated clicking. Animals that received T had significantly larger suprasternal scent glands than those that received DHT or B. The significance of these findings is discussed with respect to the development of sex differences in behavior in eutherian mammals. PMID- 8225260 TI - Behavioral depression: opposite effects of neonatal dexamethasone and ACTH-(4-9) analogue (ORG 2766) treatments in the rat. AB - Permanent changes in novelty-induced arousal and behavioral depression were studied in adult male Wistar rats having received sc injections of 1 micrograms/g body wt dexamethasone (DEX) or ACTH-(4-9) analogue (ORG 2766), or the combined treatment of these substances at Postnatal Days 1, 3, and 5. Treatment with DEX increased immobility in the Porsolt's water immersion and closed-field tests, delayed start latency, and attenuated orientation motility in an open-field, and enhanced defensive burying activity. On the contrary, the ACTH peptide caused more active behavior, resulted in an increased motility in the Porsolt's test, and decreased immobility in the closed-field chamber compared to controls. Behavioral reactivity of rats after combined DEX and ACTH peptide treatments was comparable to that of saline controls. The hormone treatments did not alter basal and stress-induced circulating corticosterone levels assessed at the adult age. The data suggest that neonatal DEX strengthens the development of brain mechanisms supporting behavioral depression in response to stressful situations, while ORG 2766 has principally an opposite effect and is able to compensate the long-term aberrant behavioral effects of neonatal DEX treatment. PMID- 8225261 TI - Estrus termination following copulation in female guinea pigs. AB - In the female guinea pig, estrus is inhibited after copulation. This study explored the role of the endocrine status of the male on the termination of the lordosis response in females. Ovariectomized females were treated with estradiol benzoate and progesterone to induce estrus and were then mated with males when the males were intact, castrated, and castrated and treated with testosterone propionate (TP). Lordosis responses were recorded prior to and following mating. Females who received an ejaculation from a castrated male remained sexually receptive longer than the females mated with intact males, or those males that were castrated and treated with TP. These results support the hypothesis that the factor(s) responsible for termination of estrus in females are found in the ejaculate of the male and are testosterone dependent. PMID- 8225262 TI - Individual differences among female rats in the timing of the preovulatory LH surge are predicted by lordosis reflex intensity. AB - In young female rats, we found individual differences in LH surge characteristics that could underlie differences in fertility throughout adulthood and aging. We modified a jugular cannulation technique, which enabled us to characterize four successive proestrous LH surges in regularly cycling female rats. During a surge, the timing and amount of LH were negatively correlated, so that late-onset surges reached a low peak, late in the day. Within a female rat, the timing of the LH surge, but not the blood level of LH, was stable from cycle to cycle. Thus, differences in onset and peak time between successive LH surges within one female were less than differences among females. Moreover, these individual differences in timing of the LH surge were positively correlated with individual differences in lordosis reflex intensity on proestrus. Females with early-onset LH surges had high average lordosis reflex intensity on proestrus. Therefore, lordosis reflex intensity is a nonintrusive behavioral biomarker for individual differences in the timing of the preovulatory LH surge. This relationship between behavior and the preovulatory LH surge may reflect a hormonal basis for variations in reproductive strategies throughout the life span. PMID- 8225263 TI - The effects of congenital hypothyroidism using the hyt/hyt mouse on locomotor activity and learned behavior. AB - The offspring of matings between hyt/hyt male mice and hyt/+ females were examined for somatic and behavioral differences. The hyt/hyt offspring displayed delayed somatic development for eye opening and ear extension relative to their euthyroid littermates. Behavioral measurement of locomotor activity indicated hyperactivity at 14 days of age and hypoactivity at 21 and 40 days relative to the euthyroid mice. Impaired swimming escape behavior and Morris maze spatial learning were observed in the hyt/hyt animals. Comparative evaluation of +/+ progenitor strain offspring having no hypothyroidism in their genetic background indicated significant differences in somatic and behavioral endpoints between the hyt/hyt and euthyroid (hyt/+, +/+) animals. These results confirm the utility of the hyt/hyt mouse for studies of the impact of congenital hypothyroidism on the functional development of the offspring. PMID- 8225264 TI - Borderline personality disorder and impulsive sexual behavior. PMID- 8225265 TI - Patient subgroups in state psychiatric hospitals and implications for administration. PMID- 8225266 TI - Informed consent in therapy. PMID- 8225267 TI - Informed consent in therapy. PMID- 8225268 TI - Self-mutilation. PMID- 8225269 TI - Supporting addiction with public funds. PMID- 8225270 TI - A preliminary study of recidivism under managed mental health care. PMID- 8225271 TI - Trends and treatment models in emergency psychiatry. PMID- 8225272 TI - Godinez v. Moran: the U.S. Supreme Court considers competence to stand trial. PMID- 8225273 TI - Spotlight on Canada. PMID- 8225274 TI - A specialized mental health plan for persons with severe mental illness under managed competition. AB - Many concerns have been raised about the special problems of providing care for severely mentally ill persons in a reformed health care system based on managed competition. The authors describe what will likely be basic features of the reformed system and discuss potential problems in serving this population. The authors recommend the development of special mental health maintenance organizations (MHMOs) that would serve only persons with severe mental illness. The MHMO would emphasize case management in the community and would provide a fixed point of responsibility for clinical care of these patients. Two methods of reimbursing MHMOs are proposed. Each region's health insurance purchasing cooperative (HIPC) could reimburse the MHMO on a capitated risk-adjusted basis. Alternatively, HIPCs could require the general health plan to operate or contract for MHMOs. In each case, the HIPC would provide quality-of-care oversight and assign a team to act as a gatekeeper for referrals to the MHMO. PMID- 8225275 TI - Poverty and the course of schizophrenia: implications for research and policy. AB - Interest in the relationship between social class and schizophrenia has diminished in recent years. However, these issues can usefully be reconsidered in light of increased recognition that schizophrenia has a protean course, that deinstitutionalization and community support of persons with schizophrenia are linked to welfare programs, and that conceptualizations and measurements of social class and its impact on the daily life of persons with schizophrenia have been rudimentary. The author examines the role that social class, especially poverty, plays in the course and outcome of schizophrenia. He reviews literature on the contributions of social structure and policy to the economic status of persons with schizophrenia, identifies variables associated with poverty that are found disproportionately among persons with schizophrenia, and examines the psychological significance of those variables. The overview is used to develop three approaches to research: examining the role of the chronic stress of poverty in the vulnerability model of schizophrenia, using poverty as a point of departure for investigation by considering persons with schizophrenia as primarily indigent rather than primarily mentally ill, and delineating how poverty and elements of schizophrenia influence each other. PMID- 8225276 TI - Access to health care for uninsured Hispanics: policy recommendations. AB - Uninsured and underinsured persons in the United States have limited access to health and mental health care and often enter the health care system via hospital emergency rooms. This pattern is even more pronounced among ethnic minorities, particularly Hispanics, a disproportionate number of whom are uninsured. The author reviews the problems faced by Hispanics in gaining access to the health and mental health care system and examines the current health and mental health status of the Hispanic population. He suggests public policy initiatives to increase the likelihood that currently uninsured members of ethnic minorities, including Hispanics, will have access to high-quality health and mental health care. PMID- 8225277 TI - Australia's national mental health policy. AB - In April 1992 the health ministers of all Australian states, territories, and the federal government endorsed Australia's first National Mental Health Policy. The major principles outlined in the policy include protecting consumers' rights, setting national service standards, mainstreaming mental health services with general health services, better integrating inpatient and community mental health services to ensure continuity of care, and linking mental health services and other social and disability services. A five-year National Mental Health Plan, accompanied by additional federal funding, has also been released, with time frames for implementing the policy in all states and territories and at the federal level. PMID- 8225278 TI - The least restrictive alternative in the postinstitutional era. AB - The legal concept of the right of psychiatric patients to treatment in the least restrictive alternative has come to mean treatment anywhere but in the state hospital, as illustrated in two large communities in Ohio and Massachusetts. Oversimplified application has led to the use of treatment setting as the only measure of restrictiveness. Patient's characteristics and needs have been ignored, along with the possibility of wide variation in program quality within each class of facility. The role of the state hospital has become uncertain, making it difficult to attract and keep qualified professional staff. The authors argue for an expanded view of the concept that includes consideration of patients' needs. They contend that the concept should not be used as a sole reason to avoid admitting patients to state hospitals or to continue the down sizing of public institutions. PMID- 8225279 TI - When there are almost no state hospital beds left. AB - The number of state hospital beds nationwide has been reduced to 40 per 100,000 population; in California, nonforensic state hospital beds have decreased to 8.3 per 100,000. The main effects of this reduction are becoming more evident. Most seriously affected have been the new generation of chronically and severely mentally ill persons who have reached adulthood since deinstitutionalization. Those who need intermediate or long-term hospitalization and cannot receive it may become even more desperate and psychotic; may avoid treatment entirely; and may turn to drug abuse and become homeless or incarcerated in jail. Moreover, the small number of long-stay beds can lead to a breakdown in the public mental health system and attempts to avoid responsibility for persons who need both acute and long-term hospitalization. This avoidance can be manifested by raising admission criteria for acute treatment or shifting responsibility to other systems. The authors recommend providing intermediate and long-stay hospitalization to the extent needed. PMID- 8225280 TI - Obtaining social welfare benefits for persons with serious mental illness. AB - The types of social welfare benefits available for mentally ill patients, including Social Security, Social Security Disability Insurance, Supplemental Security Income, Aid to Families With Dependent Children, Medicare, and Medicaid, are briefly described, and their eligibility criteria are summarized. Special requirements that may apply to immigrants are briefly noted. Case vignettes illustrate how mental health professionals can help patients obtain benefits to improve the quality of their living conditions. PMID- 8225281 TI - Strategic issues for the successful merger of residency training programs. AB - Retrenchment in federal financing, more stringent residency accreditation standards, fewer psychiatric residents, and other factors are putting increased pressure on psychiatric residency training programs to collaborate with or even merge with other residency programs. To improve the likelihood that a collaboration or merger of psychiatric residencies will work, administrators must address such issues as educational philosophy, goals of education and training, governance of the combined program, the impact on faculty and trainees, and the separate institutional cultures. Experiences from the merger of the residency programs at the Institute of Living and the University of Connecticut illustrate some of the factors. PMID- 8225282 TI - A multidisciplinary public-academic liaison to improve public mental health services in South Carolina. AB - The state of South Carolina is in the process of transforming its core services for people with severe and persistent mental illnesses to a community-based model. This paper describes the planning and on-going implementation of an alliance of academic programs, mental health advocates, and service providers forged by the state department of mental health in 1990. Goals of the consortium are to develop graduates who possess the needed knowledge, skills, and attitudes to work in the public sector; to undertake collaborative research to improve services in that sector; and to maintain the effectiveness and satisfaction of current workers in the public system. The authors describe activities of the consortium to achieve these goals and give examples of enhanced academic involvement in the public mental health system as a result of the consortium's activities. PMID- 8225283 TI - Comprehensive clinical services for substance abuse treatment in an inner-city general hospital. Substance Abuse Division, Department of Psychiatry, Bellevue Hospital, and New York University School of Medicine, New York City. PMID- 8225284 TI - Living skills training for veterans with serious mental illness--Social and Independent Living Skills Program, West Los Angeles Veterans Affairs Medical Center. PMID- 8225285 TI - Clinical and special education services for severely disturbed children- Developmental Therapy Program, Rutland Psychoeducational Services, Athens, Georgia. PMID- 8225286 TI - Specialized services for persons with medical illnesses and disabilities--The Medical Illness Counseling Center, Chevy Chase, Maryland. PMID- 8225287 TI - Community mental health outreach within a home health care agency--Community Mental Health Services, Visiting Nurse Service of New York, New York City. PMID- 8225288 TI - Carbamazepine-induced Stevens-Johnson syndrome. PMID- 8225289 TI - The new community psychiatry in 2000 A.D. PMID- 8225290 TI - Should "The chronic patient" be replaced? Reader responses. PMID- 8225291 TI - Nursing outreach to rural elderly people who are mentally ill. PMID- 8225292 TI - Treating neurotic problems that do not respond to psychodynamic therapies. PMID- 8225293 TI - Mental health services for older adults in the VA system. PMID- 8225294 TI - Impact of assertive community treatment on the use of state hospital inpatient bed-days. AB - OBJECTIVE: The purpose of the study was to determine whether use of inpatient services at a state hospital was reduced by implementation of an assertive community treatment program among persons in one of the hospital's catchment areas who were at high risk for rehospitalization. METHODS: Bed-day utilization by program participants during the fiscal year before the program was implemented (1986) and three subsequent fiscal years was compared with utilization by persons in catchment areas not served by the program. For the 66 program participants, hospital use during the year before program intake was compared with use in the year after intake using t tests. RESULTS: Utilization of bed-days by persons in the program's catchment area was reduced by 28 percent in the third fiscal year after program implementation, compared with an increase of 15 percent among persons in the hospital's other catchment areas. In the year after the program was implemented, participants were hospitalized a mean of 27.7 days, compared with a mean of 80 days in the year before the program. CONCLUSIONS: The assertive community treatment program significantly reduced use of inpatient days and improved continuity of care. PMID- 8225295 TI - Relationship between case manager contact and outcome for frequently hospitalized psychiatric clients. AB - OBJECTIVE: Case management services and client outcomes in seven programs based on the assertive community treatment model were examined to determine variability in the frequency and location of services, to determine the relationship of frequency of services with reduction in hospital use and with client satisfaction with services, and to identify subgroups of clients who received different patterns of services. METHODS: Correlational and cluster analyses were used to examine patterns of service use for 155 clients with serious mental illness who averaged 10.6 lifetime psychiatric hospitalizations. Data were drawn from four earlier studies of the treatment model. RESULTS: The programs varied substantially in the mean frequency of service provided. Contrary to expectations, service intensity was not linearly related to client outcomes. However, programs that delivered very low frequencies of service were ineffective in reducing hospital use. Finally, cluster analysis of service variables identified five client subgroups. These subgroups differed on demographic characteristics and mental health history but not on outcomes. CONCLUSIONS: A minimum intensity of services, individualized for each client, may be necessary to reduce hospital use for frequently hospitalized clients. Service utilization clusters may represent clients with different service needs and preferences. PMID- 8225296 TI - The effect of intensive case management on hospitalization of patients with schizophrenia. AB - OBJECTIVE: The study aim was to determine if a community-based psychiatric rehabilitation program with intensive case management was more effective than traditional outpatient services in reducing hospitalization of chronic schizophrenic patients. METHODS: One hundred twelve schizophrenic patients enrolled in the intensive case management program were matched by age, number of previous hospitalizations, and days spent in the hospital with 112 schizophrenic patients who received medication services and minimal case management. The two groups were compared on the amount of services used and the number of hospitalizations and hospital days over a 24-month study period. RESULTS: Patients in the intensive case management program had been enrolled for a mean of 10.96 months, with a range from one to 24 months. They used significantly more mental health services during the study period compared with the patients in the control group. However, no significant differences were found between the two groups in number of hospitalizations and hospital days. Both groups showed an increase in the number of hospitalizations over the study period. CONCLUSIONS: A community-based psychiatric rehabilitation program with intensive case management was no more effective than medication services and minimal case management in reducing hospitalization among chronic schizophrenic patients. PMID- 8225297 TI - The hostel outreach program: assertive case management for homeless mentally ill persons. AB - OBJECTIVE: This study measured the impact of an assertive case management program for psychiatrically disabled homeless persons in metropolitan Toronto. It was hypothesized that the program would improve residential stability, reduce psychiatric symptoms, improve social functioning, improve social networks, and increase use of appropriate services. METHOD: For 59 clients admitted to the program, assessments for the nine-month period before program entry were completed and were repeated nine months later. The Brief Psychiatric Rating Scale and a version of the Scale for Level of Functioning were the main measures of outcome. RESULTS: At follow-up significant improvements in residential stability and reductions in psychopathology were demonstrated. Improvements in social functioning and increases in social network size were significant. Although no baseline data about service use were collected, clients used basic support services during their first nine months in the program. CONCLUSIONS: The success of the program demonstrates that a difficult-to-treat patient population can be helped in a humane fashion if trained personnel are available. PMID- 8225298 TI - Mental and physical health of homeless persons who use emergency shelters in Vancouver. AB - OBJECTIVES: Homelessness is often associated with deinstitutionalization of chronic mentally ill people, but estimates of the number of mentally ill people in the homeless population vary. The purpose of the study was to determine the extent of psychiatric problems among the users of shelters for homeless persons in Vancouver, British Columbia. In addition, the study documented the demographic and physical health characteristics of shelter residents. METHODS: Researchers surveyed 124 emergency shelter users about their self-reported physical and mental health status and assessed their mental health status using the Brief Psychiatric Rating Scale (BPRS). RESULTS: Shelter users in Vancouver were predominantly a young, male, single mobile population. About half reported a current physical health problem, 44 percent reported use of nonprescribed drugs, and 69 percent reported use of alcohol. Nineteen percent reported a current mental or emotional problem, with schizophrenia and bipolar disorder the most common diagnoses reported. BPRS scores indicated that depression, anxiety, and tension were common problems. CONCLUSIONS: Although the number of individuals with mental illness in the population surveyed was lower than in similar populations in the United States, the presence of mentally ill people in Vancouver shelters suggests that shelters should address mental health issues as well as provide services to ensure residents' basic survival. PMID- 8225299 TI - Characteristics of veterans and nonveterans in three samples of homeless men. AB - OBJECTIVE: To uncover possible explanations for the large numbers of veterans in the homeless population, the study sought to identify differences in sociodemographic and clinical characteristics between homeless male veterans and other homeless men. METHODS: A secondary analysis of data from three surveys of homeless Americans conducted during the mid-1980s was used to compare homeless veterans and homeless nonveterans. RESULTS: Compared with nonveterans, homeless veterans were older, more likely to be white, better educated, and more likely to be previously or currently married. Generally, veterans did not differ from nonveterans on any indicator of residential instability, current social functioning, physical health, mental illness, or substance abuse. Examination of national data comparing domiciled veterans and nonveterans in the general population showed that observed sociodemographic differences between homeless veterans and nonveterans were largely explained by differences between these groups in the general population. CONCLUSIONS: Although homeless veterans had higher educational levels and were more likely to have been married in the past than homeless nonveterans, these advantages did not appear to protect them from homelessness. Veterans appear to be at risk for homelessness for much the same reasons as other American men. PMID- 8225300 TI - Implementation of the Boys Town psychoeducational treatment model in a children's psychiatric hospital. AB - OBJECTIVE: The study assessed the effect of the Boys Town psychoeducational treatment model on the therapeutic milieu of a state-operated psychiatric hospital serving children and adolescents. METHODS: Direct care staff from two units for adolescents and one unit for children were trained to implement the model, which guides staff in planned and spontaneous interactions with patients. Patients rated their satisfaction with direct care staff immediately before the model was implemented and one year later. Staff perceptions of changes in the treatment environment were assessed by retrospective interviews. RESULTS: Patients in two of the three units indicated significantly greater satisfaction with direct care staff after the psychoeducational treatment model was implemented. Results from retrospective interviews with staff showed statistically significant improvements in their perceptions of the therapeutic milieu. Staff comments recorded during the interviews supported the quantitative results. CONCLUSIONS: Training direct care staff at a children's psychiatric hospital to use the Boys Town psychoeducational treatment model significantly improved perceptions of the therapeutic milieu among both staff and patients. PMID- 8225301 TI - Clinical significance of inpatient family intervention: conclusions from a clinical trial. AB - OBJECTIVE: To test whether the statistically significant results of a randomized clinical trial of an inpatient family intervention were clinically significant for hospital practice, the authors reanalyzed outcome data using a measure of clinical significance based on the extent to which patients had recovered during the course of the intervention. METHODS: A total of 169 hospitalized subjects and their families were randomly assigned to a psychoeducational inpatient family intervention or to a comparison group. Patient and family outcome measures were assessed at admission, discharge, and six and 18 months after admission. Analyses of statistically significant differences in outcome suggested that inpatient family intervention was effective for certain patient subgroups identified by gender and diagnosis. Global Assessment Scale scores two or more standard deviations above the pretreatment (admission) mean were used as indicators for clinically significant improvement. RESULTS: The reanalysis confirmed that inpatient family intervention was associated with clinically significant improvement at discharge, especially for female patients and patients with chronic schizophrenia and bipolar disorder. These effects were maintained six months after admission before attenuating at 18 months. CONCLUSIONS: Inpatient family intervention results in clinically meaningful outcomes for certain subgroups of patients and their families. PMID- 8225302 TI - Using ECA survey data to examine the effect of job layoffs on violent behavior. AB - OBJECTIVE: The study examined two hypotheses: that being laid off increases the likelihood of violence and that being employed in an industry in which employment is unexpectedly low decreases the likelihood of violence. METHODS: Logistic regression analyses were used to examine data for more than 4,000 persons who participated in initial and follow-up interviews as part of the Epidemiologic Catchment Area survey. Data for persons who were working and not violent at the time of the initial interview but who were unemployed at reinterview were examined, as were data for persons who remained employed in industries with low employment levels. RESULTS: The risk of violent behavior of those who were laid off was nearly six times higher than that of their employed counterparts. Controlling for concurrent psychiatric disorder did not reveal a lower risk. The risk of violent behavior was reduced among those who remained employed in industries where layoffs were occurring. CONCLUSIONS: Economic contraction seems to affect violence in the community in two countervailing processes--one process increases violence and one reduces it. However, the net effect may well be that violence decreases in communities experiencing economic contraction. PMID- 8225304 TI - Insight about psychosis among outpatients with schizophrenia. PMID- 8225303 TI - A "facilitated" model of inpatient psychiatric care. AB - The authors describe a model for an effective partnership between a large health maintenance organization and a fee-for-service acute inpatient psychiatric unit. They present data from five years of experience with the model on a unit serving a catchment area of one million plan members. The model, which is based on "facilitated" care rather than managed care, emphasizes crisis intervention and a strong medical orientation. The HMO contracted with seven psychiatrists to provide treatment and helped develop a value system shared by the physicians and hospital staff. A clinician represented the HMO on the unit and played a key decision-making role in patient care. A total of 4,945 patients were admitted over five years. Costs per admission were reduced 47 percent during this period; the readmission rate was 16.9 percent. Implementation of the model resulted in the delivery of high-quality cost-effective care. PMID- 8225305 TI - The effects of smoking bans on extended care units at state psychiatric hospitals. PMID- 8225306 TI - Staff perspectives on sexual behavior of patients in a state psychiatric hospital. PMID- 8225307 TI - Oral health of state hospital patients. PMID- 8225308 TI - Supported housing. PMID- 8225309 TI - Video violence. PMID- 8225310 TI - The pseudoautosomal regions of the human sex chromosomes. AB - In human females, both X chromosomes are equivalent in size and genetic content, and pairing and recombination can theoretically occur anywhere along their entire length. In human males, however, only small regions of sequence identity exist between the sex chromosomes. Recombination and genetic exchange is restricted to these regions of identity, which cover 2.6 and 0.4 Mbp, respectively, and are located at the tips of the short and the long arm of the X and Y chromosome. The unique biology of these regions has attracted considerable interest, and complete long-range restriction maps as well as comprehensive physical maps of overlapping YAC clones are already available. A dense genetic linkage map has disclosed a high rate of recombination at the short arm telomere. A consequence of the obligatory recombination within the pseudoautosomal region is that genes show only partial sex linkage. Pseudoautosomal genes are also predicted to escape X inactivation, thus guaranteeing an equal dosage of expressed sequences between the X and Y chromosomes. Gene pairs that are active on the X and Y chromosomes are suggested as candidates for the phenotypes seen in numerical X chromosome disorders, such as Klinefelter's (47,XXY) and Turner's syndrome (45,X). Several new genes have been assigned to the Xp/Yp pseudoautosomal region. Potential associations with clinical disorders such as short stature, one of the Turner features, and psychiatric diseases are discussed. Genes in the Xq/Yq pseudoautosomal region have not been identified to date. PMID- 8225312 TI - Two novel frameshift mutations associated with the presence of direct repeats of the LDL receptor gene in familial hypercholesterolemia. AB - Two novel frameshift mutations were detected in the mutant LDL receptor genes responsible for familial hypercholesterolemia. One was a 5-bp insertion at codon 395 in exon 9, and the other was a one nucleotide deletion at codon 531 in exon 11. Both mutations alter the reading frame and consequently produce a premature stop codon in the region of the mature LDL receptor homologous to the epidermal growth factor (EGF) precursor. With regard to the mechanism responsible for the generation of these frameshift mutations, strand slipped mispairing mediated by short direct repeats is considered to be the most likely. The findings seem to support the hypothesis that a short direct repeat in DNA sequence can have a profound influence on the stability of a given gene and promote human gene mutations. PMID- 8225311 TI - Deletion of arginine (608) in acid sphingomyelinase is the prevalent mutation among Niemann-Pick disease type B patients from northern Africa. AB - There is a high incidence of Niemann-Pick type B disease in the Maghreb region of North Africa, which includes Morocco, Algeria and Tunisia. A hypothesis that there may well be a common, predominant mutant acid sphingomyelinase allele responsible for the type B phenotype in this population has been tested. A deletion of an arginine codon at amino acid residue 608 was found in one patient. The same mutation was also observed in another of our cases. An original screening procedure using 3'-end digoxigenin-labeled allele-specific oligonucleotides and chemiluminescent detection was developed and used parallel to the conventional assay with 5'-end radiolabeled oligonucleotides. Of the 15 non-related, non-Jewish North African type B patients studied, 12 were homozygous and two compound heterozygous for this deletion (26 delta R608 alleles/30 mutant alleles). Among type B patients from other geographic regions (France, UK, Italy, Czechoslovakia), this mutation was observed in only one of the 16 alleles studied. Our results indicate that deletion of arginine 608 in the acid sphingomyelinase gene is the highly prevalent mutation underlying Niemann-Pick type B disease in the population of Maghreb. A varying severity of the clinical and enzymatic expression within the non-neuronopathic phenotype has however been observed in patients homozygous for the mutation. PMID- 8225313 TI - A study of the association between schizophrenia and the dopamine D3 receptor gene. AB - A study of the genetic association between schizophrenia and a BalI polymorphism in exon 1 of the dopamine D3 (DRD3) gene, a candidate gene for schizophrenia, was conducted. The polymorphism was examined in 91 patients whose symptoms satisfied DSM-III-R for schizophrenia and 90 controls. There were no significant differences between the groups in allele frequencies or genotype counts. Contrary to a previous report, the patients were no more likely to be homozygous than controls. Moreover, no association with the presence of illness could be demonstrated when the patients were grouped according to sex, age of onset, history of admission to psychiatric institutions or positive family history. PMID- 8225314 TI - Retrospective study of the parental origin of the extra chromosome in trisomy 18 (Edwards syndrome). AB - The parental origin of the extra chromosome in trisomy 18 was traced in 30 informative families using highly polymorphic (CA) repeats mapped on the long arm of chromosome 18. Proband DNA was recovered from slides of chromosome preparations in 28 cases and from paraffin-embedded tissues in two cases. The extra chromosome was found to be of maternal origin in 26 cases (86.7%), and paternal origin in 4 cases (13.3%). PMID- 8225315 TI - Detection of confined placental mosaicism in trisomy 18 conceptions using interphase cytogenetic analysis. AB - Fluorescence in situ hybridization provides a rapid and accurate technique for detecting chromosomal aneuploidy. It is an excellent method for identifying mosaicism in placental tissues following prenatal diagnosis. Mosaicism, in the form of confined placental mosaicism, occurs im approximately 1%-2% of viable pregnancies studied by chorionic villus sampling at 9-11 weeks of gestation. It has been detected in pregnancies with both diploid and trisomic fetuses and appears to have an important effect on the intrauterine fetal survival. Using both standard cytogenetic analysis and fluorescence in situ hybridization, we have studied 12 placentas from pregnancies with trisomy 18 for the presence of chromosomal mosaicism. These included 2 that were spontaneously aborted, 5 that were terminated after prenatal diagnosis, and 4 that were delivered as either stillborn or liveborn. Significant levels of mosaicism, confined exclusively to cytotrophoblast, were detected in 7 pregnancies. This study demonstrates the usefulness of interphase cytogenetic analysis of uncultured tissues as an alternative method for the detection of mosaicism. PMID- 8225316 TI - X-inactivation pattern in carriers of X-linked retinitis pigmentosa: a valuable means of prognostic evaluation? AB - In a large family with X-linked retinitis pigmentosa 2 (XLRP2), we reexamined 7 obligate carrier females and 6 daughters of obligate carriers, whose linkage relationships suggested that they carried the XLRP2 gene. The phenotype varied from totally normal eyes through mild retinal changes to complete loss of vision. The X-inactivation analysis was carried out with the highly informative probe M27 beta on DNA from blood lymphocytes. This probe detects a locus DXS255 that is differentially methylated on the active and inactive X chromosomes. In 5 blind heterozygotes (aged 43 to 68 years), we found that the X chromosome carrying the RP2 gene was methylated and active in nearly all their cells. The opposite X inactivation pattern was found in a carrier female (aged 45 years) who gave normal findings on eye examination. Carriers with less skewed X inactivation had a less severe clinical outcome. However, we found little or no correlation between their phenotypes and the methylation status of their X chromosomes. Our results suggest that it may be possible to develop a predictive test that could identify cases with severe outcome and perhaps cases with normal outcome. PMID- 8225317 TI - Identification of an uncommon haptoglobin type using DNA and protein analysis. AB - The inherited variations in haptoglobin phenotypes are attributed to the homozygous and heterozygous combinations of three common autosomal alleles: HP*1F, HP*1S and HP*2. HP*1F and HP*1S encode polypeptides that differ by two amino acids at positions 51 and 53. The formation of HP*2 is postulated to have resulted from a breakage and subsequent reunion event at non-homologous positions of two HP*1 alleles. The most common form of HP*2 is HP*2FS in which the 5' end of HP*2 resembles HP*1F and the 3' end resembles HP*1S. Homologous crossing over between HP*2 and either an HP*1F or HP*1S allele in HP*2/HP*1 heterozygotes can change the usual type of HP*2 to three other forms: HP*2SS, HP*2FF or HP*2SF. We describe a nuclear family in which the uncommon genotype HP*2SS is one parent caused initial confusion in assigning genotypes to the rest of the nuclear family. The data demonstrate the need for a cautious approach when deducing haptoglobin genotypes from molecular analysis alone. PMID- 8225318 TI - Molecular analysis of a ring chromosome X in a family with fragile X syndrome. AB - The phenotypically normal sister of a patient affected by fragile X syndrome was referred for genetic counselling and was found to carry a mosaic karyotype 46,X,r(X)/45,X. Because the probability of the simultaneous chance occurrence of fragile X syndrome and a ring chromosome X in the same family is very low, we postulated that the breakpoint of the ring chromosome X originated in the cytogenetic break in Xq27.3 responsible for fragile X syndrome. In order to determine the relative positions of the breakpoint on the ring chromosome X and the (CGG)n unstable sequence responsible for the fragile X mutation, we used molecular markers to analyse the telomeric regions of chromosome X in this family. The results showed that the ring chromosome X was the maternal fragile X chromosome and that the telomeric deletion on the long arm encompassed the (CGG)n sequence. This suggests that the cytogenetic break in Xq27.3 is distinct from the unstable (CGG)n sequence, or that the break followed by the end-to-end fusion creating the ring chromosome was not completely conservative. Analysis of DNA markers on the short arm of chromosome X evidenced a deletion of a large part of the pseudoautosomal region, allowing us to position the genes involved in stature and in some syndromes associated with telomeric deletions of Xp on the proximal side of the pseudoautosomal region. PMID- 8225319 TI - Molecular characterization of beta-thalassemia in Hungary. AB - We have identified seven different beta-thalassemia mutations and one delta beta thalassemia determinant (the Sicilian type) in 32 members of 17 Hungarian families. The most common mutation is the IVS-I-1 (G-->A) change; its high frequency is comparable to that observed in neighboring Czechoslovakia. Additional mutations are of Mediterranean origin. One rare mutation (initiation codon ATG-->GTG) was identified as an independent mutation because of the absence of known polymorphisms in the beta-globin gene. One new frameshift at codon 51 ( C) was observed in a single individual; hematological data were as expected for a beta zero-thalassemia heterozygosity. PMID- 8225320 TI - "Premature anaphase" in a couple with recurrent miscarriages. AB - An increased frequency of mitoses with centromere separation affecting all chromosomes was found in lymphocyte cultures from a couple with recurrent spontaneous abortions. The phenomenon was observed in both the wife and husband. The abnormal behaviour of centromeres may predispose the individual to cell division errors, the consequence of which may be a spontaneous abortion. PMID- 8225321 TI - The molecular defect in propionic acidemia: exon skipping caused by an 8-bp deletion from an intron in the PCCB allele. AB - Propionic acidemia is an autosomal recessive metabolic disease resulting from a deficiency of propionyl CoA carboxylase (PCC) activity. To investigate the genetic basis of propionic acidemia, we isolated a cDNA encoding the precursor of the beta subunit of human PCC (beta PCC). The cloned cDNA sequence was 1,832 bp long and the open reading frame of 1,617 nucleotides encoded a polypeptide of 539 amino acids with a molecular mass of 58,202 Da. The human beta PCC sequence shared a high degree of homology (91%) with the full-length cDNA of rat beta PCC at the amino acid level; there were only 47 differences among 539 amino acid residues compared. Polymerase chain reaction amplification and sequencing of cDNA from a beta subunit-deficient Japanese patient revealed a deletion of 101 nucleotides consisting of one exon from mature mRNA. This deletion resulted in a frameshift and a stop codon in the new frame. Analysis of the genomic DNA revealed a homozygous 8-bp deletion from bp3 to bp10 of the intron just downstream of the deleted exon. This deletion disrupted the consensus 5' splice signal and led to exon skipping. PMID- 8225323 TI - A de novo nonsense mutation in exon 28 of the neurofibromatosis type 1 (NF1) gene. AB - We have screened a total of 105 unrelated patients with neurofibromatosis type 1 (NF1) for mutations in exon 28 of the NF1 gene using heteroduplex analysis and single strand conformation polymorphism analysis. One novel mutation has been identified and characterised. This mutation involves a 13-bp deletion (AAACTGGCTGAGC or AACTGGCTGAGCA) from base position 5077 (or 5078) to 5089 (or 5090) of the cDNA coding sequence. This alteration leads to a reading frame shift with a premature amber termination signal (TAG) at codon 1694. In addition, there is a change from lysine to threonine at codon 1693. The truncated gene product is estimated to be 1125 amino acid residues shorter than the predicted normal protein (2818 amino acids). PMID- 8225322 TI - A high-resolution cytogenetic map of human chromosome 12: localization of 195 new cosmid markers by direct R-banding fluorescence in situ hybridization. AB - We have constructed a high-resolution cytogenetic map of human chromosome 12 with 195 newly isolated cosmids by direct R-banding fluorescence in situ hybridization. The fluorescent signals of 195 clones were evenly distributed throughout chromosome 12, but sublocalized preferentially to R-positive bands. This high-resolution cytogenetic map with an average map distance of 0.73 Mb on bands can, in conjunction with a genetic linkage map, facilitate the analysis of chromosomal and molecular aberrations in genetic diseases and cancers. Moreover, the cytogenetic mapping data provide starting points for establishing contig maps with cosmid clones and yeast artificial chromosomes. PMID- 8225324 TI - Human complement C81 (C8 A) polymorphism: detection and segregation of new variants. AB - In addition to the earlier detected C81(A) rare variants A1, A2 (now A3) and B1 (now B2), six new rare variants (C81 A2 new, A4, A5, A6, M1 and B1new) are described within the polymorphism of the eighth component of human complement (alpha-gamma chain subunit). Except for A3, all rare C81 A variants are only detected by isoelectric focusing, and not by SDS polyacrylamide gel electrophoresis (PAGE), in the alpha-gamma subunit. In one individual out of approximately 700 individuals studied, a reversed position of the common allele (B vs A) was observed by SDS PAGE and the isofocusing technique. The segregation of A1, A3 and A4 could be followed in putative father/child combinations. PMID- 8225325 TI - Small frameshift deletions within the COL4A5 gene in juvenile-onset Alport syndrome. AB - Small frameshift deletions within the COL4A5 gene were identified in three Alport syndrome Italian families by non-isotopic single-strand conformation polymorphism (SSCP) screening: in family RMA, a 7-bp deletion (GGGTGAA) in exon 39; in family DGR, a 4-bp deletion (TGGA) in exon 41; in family MIB, deletion of a G in exon 50. The phenotype was characterized by juvenile-onset renal failure with sensorineural hearing loss in males, and a milder clinical pattern in heterozygous females. PMID- 8225326 TI - Homozygosity for inversion (2)(p12q14). AB - Two healthy adults, brother and sister, who are homozygotes for inv2(p12q14) are reported. As this is the first report of homozygosity for this inversion the authors ask to be informed of any further known cases. PMID- 8225327 TI - A highly informative CA/GT repeat polymorphism in intron 38 of the human neurofibromatosis type 1 (NF1) gene. AB - We describe a polymorphic microsatellite in intron 38 of the neurofibromatosis type 1 (NF1) gene. The microsatellite consists of a CA/GT dinucleotide repeat detecting 8 alleles; it has a heterozygosity of 82%. PMID- 8225328 TI - Evanescent-mode applicators (EMA) for superficial and subcutaneous hyperthermia. AB - Evanescent-mode waveguide aperture applicators are proposed for hyperthermic treatments of superficial and subcutaneous tissues. They consist of air-filled waveguide segments which are working below the cutoff frequency, and therefore support only evanescent transverse modes. These are excited by radiators of suitable symmetry and configuration to produce modal heating fields of selected cross-section. This field is emerging from the waveguide active aperture and enters the tissue to be heated through an air gap on non-critical length. These devices work in a very large range of frequencies, and are extremely simple to manufacture even with a variety of cross-section size and shape, on account of their air-filled feature. This enables to obtain good heating field flexibility with improved penetration. Their operation is safe and practical also on irregular and curved tissue surfaces. PMID- 8225329 TI - Temperature distribution in tissues from a regular array of hot source implants: an analytical approximation. AB - An approximate analytical model based upon the bioheat transfer equation is derived and used to calculate temperatures within a perfused region implanted regularly with dielectrically coated hot source implants; for example, hot water tubes, electrically heated rods, or inductively heated ferromagnetic implants. The effect of a regular array of mutually parallel heat sources of cylindrical shape is approximated by idealizing one of the boundary conditions. The solution, as could be expected, is in terms of modified Bessel functions. In calculating the temperature of each thermoregulating source in the array, the steady state power balance is enforced. The important feature of the model is that the finite size of implant diameter and its dielectric coating can be incorporated. The effect of thickness and thermal conductivity of the coating on the source and tissue temperatures along with various other interesting features are deduced from this model. The analytically calculated implant and tissue temperatures are compared with those of a numerical 3-D finite difference model. The analytical model also is used to define a range of parameters such that minimal therapeutic temperatures will be achieved in the implanted volume without exceeding prescribed maximum temperatures. This approach leads to a simple means of selecting implant spacing and regulation temperatures of hot source methods prospectively. PMID- 8225330 TI - Noninvasive measurement of current in the human body for electromagnetic dosimetry. AB - Minimally perturbing, resistive, nonferrous probes were developed for noninvasively measuring hazardous currents induced in the human body by electromagnetic fields at 1 to 200 MHz. Each probe has a resistive toroidal coil that is placed around the leg or other body member. An electrostatic shield is required to limit capacitive coupling. A new shielded test fixture provides TEM fields for calibration with a VSWR less than 1.1 from 1 to 200 MHz. A man-sized phantom was exposed to the near-field of a vertical monopole antenna at 29.9 MHz, and the value of the current measured in the leg with our probe is in reasonable agreement with measured heating. Analyses and experiments show that commercial ferrous current probes modify the circuit in which they are used, changing the current being measured. Less change is caused by our nonferrous current probes. PMID- 8225331 TI - A bidomain model with periodic intracellular junctions: a one-dimensional analysis. AB - The classical bidomain model of cardiac tissue views the intracellular and extracellular (interstitial) spaces as two coupled but separate continua. In the present study, the classical bidomain model has been extended by introducing a periodic conductivity in the intracellular space to represent the junctional discontinuity between abutting myocytes. In this model the junctional region of a myocyte is represented in a way that permits variation of junction size and conductivity profile. Employing spectral techniques, a new method was developed for solving the coupled differential equations governing the intracellular and extracellular potentials in a tissue preparation of finite dimensions. Different spectral representations are used for the aperiodic intra- and extracellular potentials (finite Fourier integral transform) and for the periodic intracellular conductivity (Fourier series). As a first application of the method, the response of a 50-cell, single interior fiber to a defibrillating current is examined under steady-state conditions. Transmembrane as well as intra- and extracellular potential distributions along the fiber were calculated. PMID- 8225332 TI - The mosaic electrical characteristics of the skin. AB - The goals of this study are: 1) to characterize the structure of the skin by measuring impedance variations with a suction microelectrode; 2) to correlate the observed impedance variations with dc current pathways through the skin; 3) to characterize the breakdown phenomenon. We constructed a suction microelectrode with a 200-microns internal diameter and performed several tests on two male subjects. Skin impedance measured from different locations on the forearm and palm varied considerably. We found that the average skin impedance on the forearm was larger than the average impedance on the palm and that the ratio between the maximal and minimal skin impedance is larger for the forearm than for the palm. For both the forearm and the palm the magnitude and variance of skin impedance decrease with increasing stimulus frequency. The density of low impedance points observed on the forearm and palm are consistent with the density of dc current pathways through the skin as indicated by traces left on 1-cm2 Ag electrodes when we passed dc current through the skin. The ratio between the highest and lowest impedances decreased as temperature decreased--at low temperatures the skin displayed mostly high impedances. We were not able to break down the skin using the suction microelectrode. The tests with dry and wet electrodes suggest that breakdown is of thermal nature, and that the thermal capacitance of the saline in the suction microelectrode prevents the temperature of the underlying skin from increasing very rapidly. In conjunction with the larger impedance values, this would tend to increase the breakdown voltage. PMID- 8225333 TI - Linear and nonlinear techniques for the deconvolution of hormone time-series. AB - Pulsatile hormone secretion is usually investigated by measuring hormone concentration in samples of peripheral plasma. In this paper, the deconvolution of hormone time-series to reconstruct the instantaneous secretion rate of glands is considered. Various techniques are discussed and compared in order to overcome the ill-conditioning of the problem and reduce the computational burden. In particular, linear techniques based on least squares, maximum a posteriori (MAP) estimation, and Wiener filtering are compared. A new nonlinear MAP estimator that keeps into account the non-Gaussian distribution of the unknown signal is worked out and shown to yield the best results. The performances of the algorithms are tested on simulated time-series as well as on series of Luteinizing Hormone (LH). PMID- 8225334 TI - Dynamic formant tracking of noisy speech using temporal analysis on outputs from a nonlinear cochlear model. AB - In this paper we take a modeling approach to studying representation of formant frequencies of spoken speech and speech in noise in the temporal responses of the peripheral auditory system. On the basis of the properties of the representation, we have devised and evaluated a cross-channel correlation algorithm and an interpeak interval analysis for automatic formant extraction of speech which is strongly dynamic in acoustic characteristics and is embedded in noise. The basilar membrane model used in this study contains laterally coupled damping elements, which are made monotonically dependent on the spatial distribution of the short-term power in the outputs of the model. Efficient digital implementation and the related salient numerical properties of the model are described. Simulation results from the model in response to speech and speech in noise illustrate temporal response patterns that are tonotopically organized in relation to speech formant parameters with little influence by the noise level. By utilizing such relations the devised cross-channel correlation algorithm is shown to be capable of accurately tracking formant movements in spoken syllables and sentences. PMID- 8225335 TI - Implementation of ultrasound time-domain cross-correlation blood velocity estimators. AB - The implementation of real-time blood velocity estimators using time-domain cross correlation is investigated. The basic algorithm is presented for doing stationary echo canceling, cross-correlation estimation and subsequent velocity estimation. Sampled data acquired at rates of approximately 20 MHz are used in the algorithm imposing a heavy burden on the signal processing hardware. The algorithm is analyzed with regard to the high sampling frequency, and a method for performing real-time high-speed data movement and cross-correlation is suggested. Implementation schemes based on using the sign of the data as well as the full precision are proposed. From an analysis of the process it is concluded that the sign data implementation can attain real-time processing. This can also be obtained for the full precision data, however, at the expense of using a number of dedicated signal processing chips. Both implementations suggested can handle the estimation of velocities for A-lines acquired from multiple directions. PMID- 8225336 TI - Mathematical analysis and computer simulation of the respiratory system in the newborn infant. AB - A mathematical model of neonatal respiratory control is proposed which can be used to stimulate the system under different physiological conditions. The model consists of a continuous plant and a discrete controller. Included in the plant are lungs, body tissue, brain tissue, a cerebrospinal fluid compartment, and central and peripheral receptors. The effect of shunt in the lungs is included in the model and the lung volume and the dead space are time varying. The controller utilizes outputs from peripheral and central receptors to adjust the depth and rate of breathing and the effects of prematurity of peripheral receptors are included in the system. Hering-Breuer type reflexes are embodied in the controller to accomplish respiratory synchronization. The model is examined and its simulation results under test conditions in hypoxia and hypercapnia are presented. PMID- 8225337 TI - Computer-optimization of vascular trees. AB - Arterial branchings closely fulfill several "bifurcation rules" which are deemed to optimize blood flow. The question is whether these local criteria in conjunction with a general optimization principle can explain the overall structure of an arterial tree. We present a model of an arterial vascular tree which is grown on the computer by successively adding terminal vessel segments. Each new terminal segment is connected to the optimum site within the preexisting tree, and the new bifurcation is optimized geometrically. After each step of adding and optimizing, the whole tree is rescaled to meet invariant boundary conditions of pressure and flow at each terminal site. Thus, local geometric optimization is used to induce simultaneously an optimized global structure. The comparison between the model and real coronary arterial trees shows good agreement regarding structural appearance, morphometric parameters, and pressure profiles. PMID- 8225338 TI - Selective stimulation of peripheral nerve fibers using dual intrafascicular electrodes. AB - We have studied activation of nerve fibers by pairs of Pt-Ir wire electrodes implanted within single fascicles of the nerve innervating the gastrocnemius muscle in cats. The purpose of this study was to determine if these intrafascicular electrodes can activate nerve fibers in different fascicles independently of each other and if they can also be used to activate separate subsets of axonal populations within a single fascicle. The average overlap of activated nerve fiber populations was 5.5% between fascicles and 27% within a fascicle, indicating that such selective activation is possible with these electrodes. PMID- 8225339 TI - Multichannel ECG data compression by multirate signal processing and transform domain coding techniques. AB - In this paper, a multilead ECG data compression method is presented. First, a linear transform is applied to the standard ECG lead signals which are highly correlated with each other. In this way a set of uncorrelated transform domain signals is obtained. Then, resulting transform domain signals are compressed using various coding methods, including multirate signal processing and transform domain coding techniques. PMID- 8225340 TI - Evoked otoacoustic emissions: nonlinearities and response interpretation. AB - Evoked otoacoustic emissions (EOAE's) are acoustic signals produced by the inner ear in response to transient acoustic stimuli and can be recorded in the external auditory canal. Since emissions can be reduced or even abolished by hearing losses of peripheral origin due to trauma and pathology, EOAE's are presently considered as the most sensitive clinical test for assessing the integrity of peripheral hearing. Recordings of emissions evoked in response to transient stimuli are contaminated by an initial artifact which in most clinical environments can be a very serious limitation. For this reason, a nonlinear estimation method is widely used (derived nonlinear response, DNLR), instead of classical synchronous averaging. Aims of this paper are: i) to justify theoretically the use of the DNLR technique by means of a simple model; ii) to compare the results obtained from the classical averaging (a linear estimation technique) with those obtained from the nonlinear DNLR technique. Some suggestions of practical interest are finally inferred from present analysis. PMID- 8225341 TI - Comments on "noninvasive estimation of cardiac output". PMID- 8225342 TI - What to expect from peer review. PMID- 8225343 TI - "I'm a dentist and I'm HIV-positive.". PMID- 8225344 TI - Attitudes, practice and employer responsiveness toward infection control as seen by certified dental assistants in Illinois. PMID- 8225345 TI - Advertising--the implications and the strategies. PMID- 8225346 TI - Beyond referrals: a smart Yellow Pages investment can bolster your bottom line. PMID- 8225347 TI - Nurse practitioner managed care for persons with HIV infection. AB - This exploratory study examined whether outcomes of care for HIV-infected patients are related to whether the patient's primary provider is a physician (MD) or a nurse practitioner (NP). Functional status, symptom occurrence, self management, health service use and patients' assessment of their care were evaluated for 87 HIV-infected patients in a teaching hospital outpatient setting. NP patients were three times as likely to report their health status as only fair or poor (odds-ratio = 3.06, p = .028), and reported significantly more unpleasant symptoms over a four-week period. Despite being in poorer health than patients cared for by physicians, NP patients functioned at comparable levels and used no more health care services than MD patients. NP patients reported 45 percent fewer problems with their care (p = 0.003). Findings suggest that more extensive use of nurse practitioners could safely enhance access to care for persons with HIV related illnesses. PMID- 8225348 TI - Current Procedural Terminology (CPT) coded services provided by nurse specialists. AB - In this exploratory study, a random sample of nurses from nine nursing specialties was surveyed to identify which Current Procedural Terminology (CPT) coded procedures they perform and how frequently they perform them. CPT codes are used universally to file claims for physician payment. The sample included 74 school nurses, 67 enterostomal nurses, 53 family nurse practitioners, 43 critical care nurses, 43 oncology nurses, 40 rehabilitation nurses, 39 orthopaedic nurses, 34 nephrology nurses and 25 nurse-midwives. Specific questionnaires were developed for each specialty with codes identified by expert panels. The number of CPT codes ranged from 233 for family nurse practitioners to 58 for school nurses. The mean number of coded services performed by individual respondents ranged from 79 (FNP) to 18 (school nurses); individual respondents performed 0 162 codes. Supervision by physicians was very infrequent. Charges to Medicare in 1988 for the coded services included in the survey were $22,793,427.34 (aggregate allowable charges). The study provides some documentation of the degree to which nurses perform the same services and procedures for which physicians are being paid. If policy makers are serious about reaching innovative solutions to the problems of quality, access and cost, everything must be "on the table," including the contributions of nurses. PMID- 8225349 TI - The NIC taxonomy structure. Iowa Intervention Project. AB - Hierarchical cluster analysis was used to construct a three level taxonomy of nursing interventions. The top level consists of six domains; the second level consists of 26 classes; and the third level consists of 357 interventions. The taxonomy is the organizing framework for the recently published Nursing Interventions Classification (NIC). This article reports the research done to construct the taxonomy and advantages for its use. The taxonomy has applications for clinicians, educators, scholars and health policy makers. PMID- 8225350 TI - The mother-child relationship in the HIV-1 positive family. AB - Qualitative data (N = 80) from a larger study of HIV-1 positive mothers have revealed mother-child relationships, that is, a strong attachment along with a bond of secrecy. The relationship is described as a strong attachment along with a bond of secrecy. Children are perceived by the mothers as sources of support as well as sources of stress in the relationship. We conclude that practice and policy decisions concerning HIV-1 positive mothers should include emphasis on maintaining the health of the mother-child dyad, providing respite to mothers and providing support for future planning. PMID- 8225351 TI - Ageism within feminism. AB - Recently there has been heightened interest in linking feminist theory and nursing science. However, neither feminists nor feminist nursing scholars have included aging women in their analyses. This paper analyzes liberal, radical and socialist feminist theories and discusses the gap within all three vis-a-vis aging women. A case is presented for including aging women within feminist thinking and recommendations for incorporating feminist theory and aging within the science of nursing are explicated. PMID- 8225352 TI - The effect of thermal biofeedback and progressive muscle relaxation training in reducing blood pressure of patients with essential hypertension. AB - In order to assess the effectiveness of the thermal biofeedback training combined with the progressive muscle relaxation therapy in the treatment of patients with essential hypertension, blood pressure decline was measured on the treatment group who had the combined thermal biofeedback and progressive muscle relaxation training (N = 11), and on the control group who had only the progressive muscle relaxation training (N = 8). Baseline blood pressure was measured four times for two weeks on both groups. For the treatment group, blood pressure was measured twice before and after each of eight sessions of thermal biofeedback training for four weeks. For the control group, blood pressure was measured every two visits to a clinic for progressive muscle relaxation self-training twice before and after the self-training. A significant decline of the systolic blood pressure by 20.6 mmHg and of the diastolic blood pressure by 14.4 mmhg was observed in the treatment group. There was a tendency for both blood pressures to increase in the control group. PMID- 8225353 TI - Healing from surgery: a phenomenological study. AB - A phenomenological study explored the experience of healing in nine post-surgical patients. Extensive interviews were conducted at one and four weeks after hospital discharge. A follow-up interview was conducted after participants reviewed descriptions of the experience. Healing included four overlapping themes: Active Participation (personal responsibility to assist in the healing process); Achieving Balance (reintegration of the whole system); Evolving Beyond (examination of goals, values and priorities); and Healing Process (change over time and connections between themes). PMID- 8225354 TI - The caregiver career. AB - The family caregiver role undertaken by spouses of persons with dementia is described as a fatalistic career process with identifiable stages. In the first or Encounter Stage, the caregivers confront the diagnosis and losses of previous lifestyle patterns and acquire home nursing skills. In the middle, or Enduring Stage, caregivers manage extensive care routines and try to cope with social isolation and their mental pain. In the final, or Exit Stage, the career is relinquished to some degree either from death of the ill spouse or institutionalization. The career perspective of caregiving provides a framework for guiding nursing research and practice. PMID- 8225355 TI - Wellness: the dialectic of illness. AB - From a review of empirical and theoretical work, a model of wellness-illness evolved. The unfolding of events associated with wellness-illness is depicted as a generic paradigm with health, disease, wellness and illness existing in a dialectical relationship. Although distinct, health-disease and wellness-illness are neither mutually exclusive nor polar opposites. Rather, they are one in the same process, acknowledging the changing person in the changing world. Wellness illness is the human experience of actual or perceived function-dysfunction through the interaction of cognitive-affective dimensions. This experience arises out of intrapersonal, interpersonal, health-disease-related and extra-personal factors. PMID- 8225356 TI - Hardiness: conceptual and methodological issues. AB - Hardiness is a personality construct that is thought to influence illness outcomes by mediating the impact of stressful life events. Hardiness research has proceeded with insufficient attention to important conceptual and methodological problems associated with this construct that have appeared in the literature. This paper analyzes and synthesizes the conceptual and methodological issues associated with the use of the hardiness construct. Implications for future nursing research are discussed. PMID- 8225357 TI - Linking family theory and practice: a family nursing program. AB - Family nursing, always a component of nursing, has been recently receiving increased attention through publications, educational programs, clinical practice settings and family nursing research. Nurses are in the best position to assist families experiencing the illness of a loved one, but often lack the skills and confidence necessary to assist with the psychosocial needs. The purpose of this paper is to describe the development and implementation of a family nursing program within a large, tertiary care facility. The program's evaluation is ongoing but early outcomes indicate that day-to-day nursing practice has been positively influenced and improved job satisfaction may result. PMID- 8225358 TI - Triangulation in qualitative research: evaluation of completeness and confirmation purposes. AB - The term triangulation has been applied to research strategies intended to serve two distinct purposes, confirmation and completeness. Many researchers who claim to have used a triangulated approach fail to provide evidence to show that their approach contributed to either confirmation or completeness of the data set. This paper reviews these distinct purposes and describes how triangulation of qualitative and quantitative methods was built into a study to achieve them. Analysis of data related to a single construct illustrates how data generated by qualitative and quantitative methods can provide both convergent validity and a broader understanding of one's subject matter. PMID- 8225359 TI - Electronic networking for nurses. AB - Electronic networks can provide access to other nurses locally and around the world via electronic mail and on-line interactions. Networks can give nurses access to information and to the data that are processed and stored in computers in their own facility and in computers at remote sites. The major international networks, Internet, FidoNet, and BITNET each offer these opportunities. In addition, Internet, the network most useful to nurses, allows access to Sigma Theta Tau's International Nursing Library, E.T.Net, and the MEDLARS family of bibliographic data bases. Taking advantage of the services and resources networks offer can facilitate nursing practice, education, and research. PMID- 8225360 TI - Sleep inquiry: a look with fresh eyes. AB - A review of nursing research into the impairment of sleep in the critically ill is presented. The author asserts that in order to fully explicate the phenomena from a nursing perspective, research is needed that focuses upon the meaning of the phenomenon to the patient. PMID- 8225361 TI - Maternal role attainment. AB - Maternal role attainment (MRA) is frequently cited in theoretical articles and research published in the nursing literature. This paper presents an historical and empirical review of MRA. Variables influencing MRA are described. New data are presented raising questions about the original conceptualization of MRA and suggesting possible discrete affective and behavioral dimensions of the construct. PMID- 8225362 TI - Gossip: does it play a role in the socialization of nurses? PMID- 8225363 TI - Peripheral T-cell reactivity to bacterial superantigens in vivo: the response/anergy paradox. PMID- 8225364 TI - Self-ignorance in the peripheral T-cell pool. PMID- 8225365 TI - Intrathymic and extrathymic tolerance in bone marrow chimeras. AB - Parent-->F1 bone marrow (BM) chimeras provide a useful model for studying self tolerance induction. When prepared with supralethal irradiation (1300 cGy) and conditioned with anti-T cell antibodies, parent-->F1 BM chimeras are devoid of host BM-derived cells; host H-2 expression is apparent in both the intrathymic and extrathymic environments but is limited to non BM-derived cells. When parent- >F1 chimeras are injected with T cells from normal parental strain mice, the expression of host H-2 antigens on nonprofessional APC might be expected to induce tolerance through induction of clonal anergy. In practice, this does not occur. Instead, a small proportion of the injected T cells is induced to proliferate and differentiate into effector cells. Tolerance is not seen. Similarly, tolerance is not apparent when thymectomized parent-->F1 chimeras are given parental strain thymus grafts. These findings suggest that the expression of host H-2 antigens in the post-thymic environment of chimeras is not intrinsically tolerogenic for mature T cells or recent thymic emigrants. Interestingly, post-thymic tolerance does occur when parental strain T cells differentiate in the endogenous thymus of chimeras. Thus, when mature CD8+ cells are prepared from thymus vs lymph nodes (LN) of parent-->F1 chimeras, tolerance to host class I antigens is more marked in LN than thymus; this applies to cytotoxic T lymphocyte (CTL) precursors, generated by limiting dilution analysis. It would appear therefore that many of the host-reactive CTL precursors generated in the thymus of chimeras undergo tolerance induction (deletion or irreversible inactivation) in the post-thymic environment. We suggest that such tolerance is a reflection of a covert form of tolerance induced in the thymus: intrathymic contact with host antigens on thymic epithelial cells (TEC) in chimeras does not delete typical CTL precursors, but these cells are rendered "semi-tolerant". When cultured in vitro in the presence of lymphokines, the cells are able to recover and differentiate into CTL. In vivo, however, the cells recognize antigen in the periphery in the relative absence of lymphokines and the cells die. Although host class I expression on TEC in chimeras deletes only a small proportion of CTL precursors, contact with TEC induces strong tolerance of CD8+ cells in terms of helper-independent proliferative responses in vitro and induction of lethal graft versus-host disease in vivo. We postulate that these latter responses are controlled by high-affinity T cells, whereas typical CTL generated in LDA are predominantly low-affinity cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8225366 TI - Molecular regulation of the IL-2 gene: rheostatic control of the immune system. AB - The delivery of costimulation and the effects of the anergic state impinge on IL 2 production via different molecular mechanisms. The strongest experimental support at this stage suggests that CD28 signaling effects mRNA stability of several lymphokine genes including IL-2. While there may also be transcriptional effects of CD28 signals in human cells, controversy surrounding relevant TCR mimics must be addressed. In the case of clonal anergy, however, transcriptional non-responsiveness is evident when anergic cells are restimulated with TCR and costimulatory signals. This repression affects predominantly AP-1 activity. So far, the nature of the repression has not been identified. PMID- 8225367 TI - Tolerance to primarily vascularized allografts in miniature swine. PMID- 8225368 TI - Effector T-cell induction and T-cell memory versus peripheral deletion of T cells. PMID- 8225369 TI - The role of thymic epithelium in the establishment of transplantation tolerance. AB - From experimental observations on induction of transplantation tolerance, we discuss a model that accounts for tissue-specific tolerance to antigens not expressed inside the thymus. It is postulated that antigens presented to differentiating T cells by thymic epithelium (or at large within the thymic environment) positively select and activate self-reactive T cells. A developmental program and/or prevalent conditions in the thymic environment restrict the proliferative potential and the class of effector functions that can be exerted by differentiating T cells activated in the thymus. These do not mediate inflammatory or cytolytic activities, but instead will produce the appropriate mediators to inhibit aggressive effector activities by other T cells activated in their proximity. Such "regulatory" functions will be locally expressed at the periphery upon recognition of tissue antigens shared with the thymus, towards newly formed thymic emigrants directed at tissue-specific antigens expressed by the same "target" cells. This mechanism imposes "dominant tolerance", based on specific self-recognition and predominantly established in the embryonic and neonatal period. Throughout life, the process of thymic positive selection results in all newly-formed T cells being susceptible to such suppressive mechanisms, but becoming increasingly refractory with time in the resting, post-differentiative stage. Absence of antigen (nonself) in the embryonic and neonatal life therefore allows for the accumulation of such "suppression-resistant" antigen-reactive T cells that will mount aggressive responses upon antigenic exposure. Tolerance or immunity thus represent two classes of specific immune responses, the relative predominance of which is determined by the frequency of each type of effector T cell, representing the antigenic overlap between thymic and peripheral tissues, as well as the frequency of tissue-specific T-cell generation, and the kinetics of peripheral antigenic exposure. Tolerance induced by hemopoietic cells to all other tissues is also "dominant" and based on thymic colonization and persistence of antigenic cells, with the consequent positive selection of regulatory T cells and peripheral conditions for the establishment of suppression. Upon this simple model, that ensures "interclonal class regulation" by "bridging" regulatory and effector T cells through the recognition of different antigens on the same target cell, other mechanisms which are based on V-region interactions among T cells (Ben-Nun et al. 1981, Pereira et al. 1989, Webb & Sprent 1990, Gaur et al. 1993) might well operate to ensure "dominant tolerance" by self-reactivity and class regulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8225370 TI - Cellular tolerance as a dynamic state of the adaptable lymphocyte. AB - The regulation of immunological tolerance is considered from the perspective of contextual discrimination, rather than self-nonself discrimination. According to the adaptive lymphocyte hypothesis, the scale of immune aggression versus tolerance can be regulated at the cell population level, but individual cells also tune and update their responsiveness under the influence of recurrent signals. The generation of a sizeable conventional immune response, which is transient and aggressive, depends critically on the perturbation to the system, which is related to the rate of appearance of the immunizing agent. These characteristics are explained in quantitative terms by the "balance of growth and differentiation model". Strong perturbations are typically associated, physiologically, with acute infections. Full activation of individual lymphocytes also requires strong metabolic perturbations, where the perturbation is defined as a measure of variation in the intensity of stimulation. Cells that fail to be activated in this way may be driven into a state which formally conforms to the operational definition of anergy. This state is characterized by a variable degree of resistance to the stereotypic mode of activation for which the cell has been programmed before. While in this state, the cell interacts with its environment: these interactions promote its viability, update its activation thresholds and its excitability, and may reprogram the cell for a different mode of response when activated later. In addition, cells engaged in such interactions may mediate context-dependent immunological functions. The characteristics of the interactions involving such anergic cells are discussed in semi-quantitative terms with the help of the "tunable activation-thresholds model". Several aspects of immunological tolerance are interpreted in a unifying way based on this conceptual framework. It is suggested that progress in our ability to evaluate and manipulate the regulation of immunological tolerance would require a methodology to conjoin many pieces of data together and to look for patterns. PMID- 8225371 TI - Cytokines and peripheral tolerance to alloantigen. AB - The induction of peripheral tolerance to alloantigen is accompanied in many cases by a decrease in the production of cytokines such as IL-2 and IFN gamma, yet a sustained production of cytokines such as IL-10 and IL-4. Whether or not this altered pattern of cytokine production in tolerant animals is causally related to the induction and/or maintenance of the tolerant state has yet to be fully determined, although experiments blocking selectively the action of IL-2 with CD25 antibodies suggest that manipulation of cytokine production may at least be a route to tolerance. Alternative methods for directly influencing the cytokine balance are sought and recent experiments on the CD28/CTLA-4-B7 interaction suggest a possible approach. PMID- 8225372 TI - Ir1 genes, peripheral cross-tolerance and immunodominance in MHC class I restricted T-cell responses: an old quagmire revisited. AB - Previous studies have shown that peripheral tolerance of one MHC molecule may influence the ability of an animal to respond to a particular antigenic determinant in the context of another MHC molecule (cross-tolerance). We describe here an investigation of the extent to which the presence of MHC molecules other than the one involved in Tc cell recognition affected whether or not a particular "antigen" (MHC/peptide) was recognized during the response of mice to infection with the flavivirus, West Nile (WNV). WNV-immune Tc cells from intra-H-2 recombinant mouse strains and F1 hybrid animals were used. In general, the antigens identified as immunogenic for five prototype mouse strains were immunogenic wherever the F1 or recombinant animal possessed the required MHC gene, but there were several notable exceptions. Firstly, while no responses were associated with Kd and Dk in BALB/c (H-2d) and CBA/H (H-2k) mice, respectively, in C3H.OH mice (Kd, Dk) both of these MHC molecules were associated with detectable responses. Secondly, in B10.A(2R) (Kk, Db) mice responses restricted by Db were not found whereas they were present in B6 mice (Kb, Db). This phenomenon was similar to previously reported phenomena with H-Y, Sendai, influenza and VV. However, it differed in two important ways. Firstly, we have no definitive evidence as yet that peripheral cross-tolerance to Kk influences responses against Db plus flavivirus as it does with H-Y and VV. Secondly, in addition to Kk, genes in the H-2s, H-2q and H-2d haplotypes influence responses to Db plus flavivirus but not responses to the other "antigens" listed above. Alternatively, the data are compatible with the concept of "immunodominance" based on a hierarchy of affinities of TCR's. the responding T-cell population appeared to focus the immune response on a limited number of "antigens"; in the presence of certain more strongly immunogenic "antigens", responses to other apparently more weakly immunogenic "antigens" were not seen. PMID- 8225373 TI - Peripheral tolerance as a multi-step mechanism. PMID- 8225374 TI - Chronic allograft rejection. PMID- 8225375 TI - Chronic rejection in experimental cardiac transplantation: studies in the Lewis F344 model. AB - Despite recognition of chronic vascular injection by numerous investigators since the beginning of experimental and solid organ transplantation, the pathogenesis of graft arteriosclerosis remains poorly understood. We have defined a reproducible model of this disease by transplanting heterotopic cardiac grafts across minor histocompatibility barriers using inbred strains of rat. We found that long-term surviving Lewis grafts in untreated F344 recipients are subjected to a chronic rejection process which results in the development of diffuse graft arteriosclerotic lesions, indistinguishable in appearance from those seen in human cardiac grafts. Immunohistochemical studies confirm that end-stage lesions are similar in composition to human lesions, made up predominantly of vascular smooth muscle cells with occasional monocytes and T cells. Analysis of serial rejecting allografts demonstrates that a distinct inflammatory stage precedes smooth muscle cell accumulation in areas of intimal thickening, suggesting that mononuclear cells play a role in the developing lesion. Endothelial expression of class II and ICAM-1 appears to lead to early mononuclear cell adherence to the endothelium. Analysis using quantitative RT-PCR confirms that MCP-1 is expressed by ED1-positive monocyte/macrophages in rejecting cardiac grafts, suggesting that this chemoattractant may help drive mononuclear cell accumulation in the expanding intima. Immunohistochemical labelling of PDGF, TNF, and IL-1 in vascular lesions suggests that these growth factors may trigger intimal vascular smooth muscle cell proliferation in chronically rejecting allografts. Hypercholesterolemia did not enhance the severity of lesion development in long term surviving allografts, suggesting that lipid levels are not a major etiologic factor in graft arteriosclerotic lesion formation in the Lewis-F344 model. Finally, the dietary manipulation of EFAD reduced graft infiltration by mononuclear cells and markedly diminished arterial lesion development in chronically rejecting grafts. Heparin anologues have previously been shown to inhibit proliferative vascular smooth muscle cell lesions in rats following endothelial injury (Clowes & Karnovsky 1977), and we are currently assessing the role of heparin in the therapy of graft arteriosclerosis in the Lewis-F344 model. We are also investigating the role of CD4-positive mononuclear cells in the pathogenesis of lesion development, using the anti-CD4 monoclonal antibody BWH-4 to deplete recipients of CD4-positive cells (Sayegh et al. 1991). In summary, our studies in the Lewis-F344 model suggest that monocyte/macrophages play an important role in the pathogenesis of cardiac graft arteriosclerosis. Future studies utilizing this model should help further elucidate the mechanisms resulting in--and help define potential therapies for--chronic rejection in cardiac transplantation. PMID- 8225376 TI - Pathogenesis and treatment perspectives of chronic graft rejection (CVR). AB - Chronic rejection is a major threat towards the long-term function and survival of transplanted hearts and kidneys. It is characterized by a proliferative remodelling of the graft vessels along with structural changes of the parenchyma and gradual deterioration of graft function. The pathogenesis is complex and multifactorial. Since grafts with chronic rejection are also subjected to a more or less intense invasion of immunoreactive cells, an important primary objective is to optimize the immunosuppressive treatment. There is no established means of prevention or treatment of chronic rejection. Pharmacological agents interfering with prostaglandin metabolism have been tried most frequently and preliminary results are also available from the use of polyunsaturated fatty acids of the omega-3 series and of heparin derivatives. Based on experimental studies the somatostatin analogue angiopeptin seems very promising today. There will certainly be an increased interest in the use of lipid-reducing agents in the future as well as antioxidant agents acting against the effects of reactive oxygen radicals and oxidative modification of LDL fractions. A strong novel candidate is carvedilol, exerting both antihypertensive, antioxidant and antiproliferative properties. PMID- 8225377 TI - A psychiatric evaluation of referred cancer and medical patients: a comparative study. AB - We evaluated the psychiatric status of referred cancer patients (N = 39) and referred medical patients at two other institutions (N = 64, N = 62). Depression, anxiety, psychosis, delirium occurred with comparable frequency in both groups. Cancer patients presented with somatic complaints more frequently. However the average number of referrals of cancer patients to the psychiatric service was very low (3/months) compared to referral of medically ill patients (32/month, 10/month). The data suggests that a comparable level of psychological distress exists in cancer patients, and psychiatric referral may help in its early recognition and alleviation. PMID- 8225378 TI - Delay in specialist consultation in cancer patients. AB - A psychiatric evaluation of 75 cases of different types of cancer revealed significant psychological morbidity in 81% of cases. The time lag, if any, between primary diagnosis and presentation to oncologists was noted and the reasons for the delay were ascertained. 62% of patients (N = 47) delayed three months or more and this group also showed significantly greater (p < 0.05) psychological morbidity compared to those who delayed less than three months (N = 28). 89% of patients used denial as a mental defence mechanism and there was no significant difference in the degree of denial displayed by both groups. Psychological reasons such as denial of the illness, fear of cancer, fear of its treatment, and domestic difficulty were commonly cited as causes of delay. It is hoped that addressing these issues will reduce delay and help early detection in cancer programmes. PMID- 8225379 TI - Pattern of cancer in Jammu region (hospital based study 1978-'87). AB - 2681 histologically confirmed cases of cancer (1402 males and 1279 females) were seen during the ten year period 1978 to 1987 in Pathology Department, Government Medical College, Jammu. The relative frequencies of cancer at various primary sites have been determined with respect to age, sex and religion. The majority of cancers were seen between 31-60 years (67.7%) with a peak in 41-50 years age group. Out of the total cancer patients 2,437 were Hindus (90.8%) and 244 were Muslims (9.2%). The most common cancer sites among males were lung, skin, larynx, prostate and lymphnodes. Cancer cervix was the most common tumor in females, followed by cancer breast, gall bladder and uterus. Among Muslims, cancer cervix and penis were infrequent. Cancer lung and larynx were also relatively less frequent in Muslims. PMID- 8225380 TI - L-asparaginase related hyperglycemia. AB - L-asparaginase is a valuable chemotherapeutic agent used in the induction of remission and improvement of long term survival in patients with acute lymphoblastic leukemia. Hyperglycemia is a well known side effect of L asparaginase. Fourteen patients developed hyperglycemia during induction therapy of acute lymphoblastic leukemia with L-asparaginase, prednisolone, vincristine and daunorubicin. Hyperglycemia was observed after a mean of five doses of L asparaginase (range 2-10). Seven of fourteen patients had neutropenic related infective episodes. Hyperglycemia resolved in all patients within 12 days (range 4-25) and two patients died of neutropenic septicemia. During reinduction therapy with the same drugs, only one out of ten patients developed hyperglycemia E-coli L-asparaginase was replaced by Erwinia asparaginase in two patients one of who had recrudescence on further therapy. Close monitoring during L-asparaginase therapy for hyperglycemia will enable prompt recognition and early correction and prevent delay in therapy of acute lymphoblastic leukemia. PMID- 8225381 TI - Recurrent ameloblastoma of the mandible and maxilla. AB - Sixteen cases of recurrent ameloblastomas of the mandible and maxilla seen over a 14 year period at the Tata Memorial Hospital were analysed with emphasis on recurrence patterns, type of primary surgery and surgical management. Seventy five percent of the cases had conservative surgery at the first instance. In our experience, recurrent tumors could be excised widely even in unusual locations with good results. PMID- 8225382 TI - Primary papillary adenocarcinoma of the seminal vesicle. AB - This paper describes a papillary adenocarcinoma solely confined to the seminal vesicle, in a 26 year old male. Imaging techniques and operative findings revealed an enlarged seminal vesicle, harboring a grossly papillary and friable tumor. Strict criteria for diagnosis have been applied, in this unusual case. The young age and unusual location of this papillary adenocarcinoma prompted us to report this case. PMID- 8225383 TI - Infantile neuroblastoma: 10 year TMH experience. AB - The results of 19 children with neuroblastoma under one year of age treated at the Tata Memorial Hospital between 1981-1990 were analysed. Biologically, neuroblastomas in children under the age of 12 months have an entirely different prognosis as compared to older children. There were four children less than six months of age and 15 children between six and 12 months. All patients with stage II (2/2) disease are alive; 3/4 patients with stage IV-S disease are disease free; 3/5 stage III patients are disease free and 2/8 patients with stage IV disease are disease free. The survival of infants with stage II and IV-S is better than for those with stage III and IV disease. PMID- 8225384 TI - Sellar-suprasellar primary malignant lymphoma: case report. AB - A patient who came with compression of the chiasma and later on had involvement of the cavernous sinus is presented. Radiological examination showed a seller, suprasellar and parasellar tumor. Histopathological examination revealed a primary Non-Hodgkins lymphoma. PMID- 8225385 TI - Primary squamous cell carcinoma-in-situ of the endometrium--a case report. AB - A case of primary squamous cell carcinoma in situ of the endometrium is reported here. The in-situ nature of the tumor, with involvement of the surface mucosa and extension into superficial endometrial glands and its incidental association with prolapse uterus are notable features. PMID- 8225386 TI - Human complement factor B: functional properties of a recombinant zymogen of the alternative activation pathway convertase. AB - The human complement factor B is a centrally important component of the alternative pathway activation of the complement system. Here we report the isolation, characterization and eukaryotic expression of the first full length cDNA transcript for human factor B. In a factor B dependent haemolysis assay, the recombinant human factor B generated by transient COS cell transfection was shown to reconstitute haemolytic activity of factor B depleted human serum. To study the biological activities assigned to factor B, the availability of recombinant polypeptides representing definite portions of the human factor B molecule is desirable. PMID- 8225387 TI - Deregulated c-fos modulates IgG2b production of B cells mediated by lipopolysaccharide. AB - We have examined effects of the deregulated c-fos protein on IgG2b production of B cells cultured with lipopolysaccharide (LPS) using splenic B cells from a transgenic line carrying the mouse c-fos gene under the control of the interferon alpha/beta (IFN) inducible Mx promoter (Mx-c-fosD). High c-fos expression was induced in the Mx-c-fosD B cells during the first two days of culture. DNA synthesis and IgG2b production were augmented in the culture. When IFN was added together with LPS, the high c-fos expression was prolonged until day 3 of culture. IgG2b production was remarkably suppressed. However, the production was not suppressed by upregulation of c-fos via exogenous IFN on day 4 of culture. These results suggest a regulatory effect of the c-fos protein on the differentiation of B cells to IgG2b producing cells at a distinct period. PMID- 8225388 TI - Inhibitor of C1q secretion suppresses the macrophage response to lipid A for nitric oxide but not for TNF production: evidence for a role of C1q in autocrine binding of TNF. AB - Studies were designed to further define the modulatory role of complement subcomponent C1q in macrophage activation by Lipid A to mediate production of TNF and cytotoxic nitric oxide (NO). Pretreatment of macrophages for 24 h with 2.5 mM 3,4,dehydro-D,L-proline (DHP), an inhibitor of C1q secretion, suppressed their response to Lipid A activation for cytotoxicity of P815 tumor targets which correlated with a corresponding decrease in NO production. In contrast, DHP pretreated macrophages displayed an increase in the release of TNF in response to Lipid A as compared to untreated controls. Time kinetic studies indicated that DHP-pretreated macrophages produced higher sustained levels of TNF activity during 1 to 24 h culture with Lipid A than did untreated control macrophages. This was confirmed by increased TNF mRNA expression in response to Lipid A by DHP treated cells. DHP-pretreated macrophages had reduced levels of cell surface C1q as determined by cytofluorometric analysis of the binding of FITC-labeled anti C1q, F(ab')2. Macrophages were also found to have reduced binding capacity for phycoerythrin-labeled rTNF (PE-TNF) by cytofluorometric analysis following DHP treatment. Exposure of DHP-pretreated macrophage to soluble C1q at 4 degrees C restored their reduced binding of PE-TNF. C1q was confirmed to bind to macrophages at 4 degrees C as detected by FITC anti-C1q, F(ab')2 and such C1q binding promoted a corresponding increased binding of PE-TNF. Macrophages which were plated over immobilized C1q were also markedly enhanced in their binding of PE-TNF probe. Our results indicate that the inhibition of macrophage secretion of C1q by DHP pretreatment, was accompanied by an increased TNF mRNA expression and release with a decrease in NO generation following Lipid A activation. Since TNF binding to DHP-treated macrophages was reconstituted by the binding of exogenous C1q to the cells, it appears that C1q may be involved in the modulation of autocrine binding of TNF for subsequent generation of cytotoxic NO. PMID- 8225389 TI - In vivo modulating effects of bacterial peptidoglycans on PHA-induced responses of porcine PBL and splenocytes. AB - The modulating effect of Brevibacterium divaricatum cell wall derivatives, i.e. peptidoglycan monomer (PGM) and peptidoglycan polymer (PGP) on phytohemagglutinin (PHA)-induced responses of peripheral blood lymphocytes (PBL) and spleen lymphocytes (SPL) from neonatal pigs farrowed to gilts or sows was tested. Both, PBL from progeny of sows and SPL from progeny of gilts responded significantly lower (P < 0.01) when PGM (14 mg/kg) was given by intraperitoneal (IP) route at Day 0 and Day 7 after farrowing as compared to the respective controls. When the same amount of PGM was given i.p. at Day 7 and Day 14, the responses of PBL and SPL from offspring of gilts were much higher (P < 0.01) than those of nontreated controls. The treatment with PGP (2.5 mg/kg) by the same route at Day 7 and Day 14 strongly stimulated (P < 0.01) the responses of PBL and SPL of pigs originating from litters of either gilts or sows, respectively. PMID- 8225390 TI - Cytokine production of the human monocytic cell line Mono Mac 6 in comparison to mature monocytes in peripheral blood mononuclear cells. AB - Mono Mac 6 is a human monocytic cell line with several features of mature blood monocytes such as CD 14 antigen expression, phagocytotic ability, and the functional ability to produce cytokines. This line is often used as an in vitro model to demonstrate the actions of monocytes. In our study, the production of cytokines by Mono Mac 6 cells in response to various stimulants was analyzed and compared to that of mature monocytes in peripheral blood mononuclear cells (PBMC). Interestingly, the Mono Mac 6 cells produced IL-1 alpha/beta, IL-6, and TNF-alpha after induction with the lectin phytohaemagglutinin A (PHA), mainly known as a T cell activator. The amount of cytokine release did not decrease in the presence of polymyxin B (Pmb), an inhibitor of LPS-induced effects. Kinetic studies revealed maximum cytokine levels 24h after stimulation, whereas human PBMC produced higher yields of all cytokines and enhancement was observed up to 48 hours after induction. Stimulation with the superantigen derived from the supernatant of mycoplasma arthritidis (MAS) induced expression of IL-1 beta, IL 6, and TNF-alpha, whereas staphylococcus enterotoxin B (SEB) did not induce any cytokine release. Further experiments analyzed the ability of Mono Mac 6 cells to produce IFN-alpha which is an important characteristic of mature monocytes. The cells were induced either with inactivated Newcastle Disease Virus (NDV), Sendai Virus, or the synthetic stimulus poly I:C IFN-alpha expression was not detected on the transcriptional or the protein level. In addition, no co-expression of IL 1 and IL-6 was observed in response to these stimuli. Since NDV, Sendai Virus, and poly I:C represent strong IFN-alpha inducers in peripheral blood monocytes, these data indicate that Mono Mac 6 cells lack the ability to express IFN-alpha. In conclusion, our findings show that this cell line is a potent cytokine producer, but the capacity to produce IFN is apparently deficient. PMID- 8225391 TI - Long-term in vivo effects of rapamycin on humoral and cellular immune responses in the rat. AB - Rapamycin (RAPA) is a strong immunosuppressant and is able to prevent allograft rejection in animal models. We have demonstrated that RAPA could strongly inhibit in vitro immunoglobulin (Ig) production by human lymphocytes. The present study investigated the long-term in vivo effect of RAPA on humoral and cellular immune responses, and the effect of RAPA on accelerated rejection. It was shown that RAPA strongly inhibited antigen (Ag) specific antibody (AB) production (i.e. cytotoxic Ab to donor lymphocytes and Ab to tetanus toxoid) during the period of drug administration. The accelerated rejection of cardiac allografts in presensitized animals was alleviated by RAPA administration. These results suggest the potential application of RAPA in treatment of presensitized candidates for organ transplantation. A little more than two months after the drug withdrawal, the rats were basically competent in Ab response to further Ag challenges. When tested 4 months after the RAPA-treatment, the rats showed uncompromised cardiac allograft rejection, and the cellular immune response in vitro according to mixed lymphocyte reaction (MLR) and mitogen-stimulated proliferation were not hampered. Such results suggest that the immune system can return to normal status without sequelae after discontinuation of RAPA. PMID- 8225392 TI - Mechanism of liposome adjuvanticity: an in vivo approach. AB - The reputation of liposomes as adjuvant of the immune response is now firmly established despite the lack of information on the mechanisms involved in their immunopotentiating properties. The rapid targeting of massive doses of antigenic material to antigen-presenting cells, especially macrophages has, however, often been invoked as the principal source of liposomal adjuvanticity. In order to test this hypothesis, we analyzed the humoral response to antigen encapsulated in liposomes containing increasing amounts of surface-exposed mannose residues, ligand specific of an exclusive macrophagic receptor. Using BSA as a model antigen, we demonstrated that the humoral response is profoundly affected by mannosylation, being of prolonged duration and either inhibited or activated depending on the immunizing doses. These results suggest that the rapidity of antigen targeting is not the sole reason to liposome adjuvanticity and that the role of liposomes as antigenic depot is probably important to sustain substantial activation through successive restimulations. In this context, the increased rapidity in antigen targeting which favors the concentration of activation signals in time, results in an under-optimization of the response at high immunizing doses and in an optimization of this response at doses that would otherwise give rise to signal of sub-threshold intensity albeit during a longer period of time. PMID- 8225393 TI - Differences in antigen-binding affinity caused by a single amino acid substitution in the variable region of the heavy chain. AB - Site-directed mutagenesis has been used to examine the importance of histidine-99 in the VH CDR3 region of a mouse/human chimeric anti-TAG72 antibody, cB72.3-1-3. The expression vectors mpSV2neo-EP1-V-m4-10C gamma 1, containing seven different mutant VH region fragments (Vm4-10) in association with the immunoglobulin enhancer (E), promoter (P1) and human genomic C gamma 1 region fragments, were transfected into a heavy chain loss mutant cell line B72.3 Mut(K), respectively. Mutant chimeric antibodies cB72.3m4-10 were purified from the transfectant supernates, and their binding affinities for the TAG72 antigen relative to that of the original cB72.3-1-3 antibody were compared. Substitution of histidine-99 by glutamine resulted in a higher affinity antibody (cB72.3m4) whereas substitution by isoleucine resulted in a lower affinity antibody (cB72.3m9). The binding affinity of these mutant antibodies varied nearly eight-fold. It was concluded that the residue at position 99 in the VH CDR3 region is in a 'contact' position in the B72.3/TAG72 antibody-combining site. The polar side-chains of glutamine and asparagine or the ionized side-chains of histidine, arginine or glutamic acid contribute to higher binding affinity, whereas the hydrophobic side chains of isoleucine, leucine or phenylalanine resulted in a lower binding affinity for the TAG72 antigen. PMID- 8225394 TI - Vitamin D-binding protein (group-specific component) is the sole serum protein required for macrophage activation after treatment of peritoneal cells with lysophosphatidylcholine. AB - In vitro treatment of mouse peritoneal cells with 1 micrograms lysophosphatidylcholine (lyso-Pc)/mL in serum free-0.1% egg albumin-supplemented RPMI 1640 medium for 30 min, followed by 3 h cultivation in a medium supplemented with human serum, resulted in a greatly enhanced Fc-receptor-mediated phagocytic activity of macrophages. Vitamin D-binding protein (group-specific component [Gc]) of alpha 2-globulin fraction was shown to be the sole serum glycoprotein required for the generation of a potent macrophage-activating factor. When a mixture of lysophosphatidylcholine (lyso-Pc)-treated nonadherent and adherent cells were cultured in a medium supplemented with a small amount of purified Gc protein (1 ng/mL), a greatly enhanced activation of macrophages was demonstrated. The generation of macrophage-activating factor from purified Gc protein was far more efficient than that from whole serum, indicating that a serum component is inhibitory to the activation process of macrophages. While three other major serum glycoproteins (alpha 2-macroglobulin, alpha 2-HS-glycoprotein and haptoglobin) were neither stimulatory nor inhibitory to lyso-Pc-primed macrophage activation, serum albumin (competitively with Gc protein) appeared to be inhibitory to the process of macrophage activation. PMID- 8225395 TI - Cytokine production in response to Epstein-Barr virus infection of peripheral blood mononuclear cells in vitro. AB - To obtain a better understanding of the immune response to Epstein-Barr virus (EBV), we measured the cytokines tumour necrosis factor (TNF)-alpha/beta, interleukin-2 (IL-2), interferon-gamma (IFN-gamma), IL-6 and granulocyte macrophage colony-stimulating factor (GM-CSF) in the conditioned medium of peripheral blood mononuclear cells from 10 healthy adults before and at 48 h and at 1, 2, 3 and 4 weeks following infection in vitro with EBV. Cultures were examined for regression of outgrowths of nascent virus-transformed B cells, and populations of cells in the cultures were analysed by flow cytometry. TNF alpha/beta was not detected in infected or non-infected cultures. In infected cultures assayed at the nominated times, the highest levels of IL-2 were detected at 48 hours, IFN-gamma at 1 week, IL-6 at 2 weeks and GM-CSF between 2 and 4 weeks. IL-6 and GM-CSF, but not IL-2 or IFN-gamma, were detected in non-infected cultures but at lower levels than in infected cultures. Nine of the 10 healthy adults showed regression of outgrowths of virus-transformed B cells and, of these, seven had antibodies to the EBV capsid antigen (VCA). Strong regression was associated with sequential increases in IL-2, IFN-gamma, and low levels of IL 6 and GM-CSF. Absent or weak regression was associated with an undetectable level of IL-2, a low level of IFN-gamma, high levels of IL-6 and GM-CSF and an increased frequency of cells bearing the phenotype CD20 and HLA-DR in the final weeks of culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225396 TI - Interleukin-6 production during chronic experimental infection. AB - The appearance of interleukin-6 (IL-6) in serum of mice was monitored during the course of chronic infection with either Brucella abortus vaccine strain 19 or a virulent Mycobacterium avium Complex (MAC) isolate. Serum IL-6 during brucella infection was higher than during infection with MAC, despite similar numbers of bacteria. Furthermore, IL-6 titres decreased after the peak of infection, falling to baseline levels before these chronic infections were eradicated. The ability of peritoneal cells or spleen cell suspensions to produce IL-6 under either specific or non-specific stimulus was greatly enhanced by infection. While production of IL-6 by these cultures was apparently mostly independent of T cells, T cells from infected mice could produce an IL-6 response. Thus CD4+ T lymphocytes prepared from mice which had recovered from B. abortus infection, cultured with antigen and antigen presenting cells, resulted in IL-6 production, which was not observed in similarly cultured CD8+ T cells, indicating a role for T cells. PMID- 8225397 TI - CD45 isoform transitions on multinegative human thymocytes differentiating in vitro mimic patterns predicted for selective events in vivo. AB - Experiments were designed to test the idea that the patterns of CD45 isoform expressed by differentiating human multinegative (MN) thymocytes (CD3- 4- 8- 19-) are regulated by the type of growth or activation stimuli delivered. We have proposed that within the thymus, CD45RA expression is fundamental to maintenance of the thymic generative lineage while a transition to CD45RO indicates entry into the path of intrathymic death. It seems likely that the signal transduction pathways leading to positive selection are different from those leading to negative selection. Upon culture, MN thymocytes proliferate in response to interleukin-2 (IL-2) or anti-CD2/28 and differentiate as defined by acquisition of CD3 as well as CD4 and/or CD8. Even at day 9 of culture, 30-40% of cells remain multinegative. DNA analysis indicates that both CD3- and CD3+ cells are actively cycling. Although the CD3- set includes a substantial number of cycling cells, it continues to express almost exclusively the CD45RA isoform. Among CD3+ progeny, 53-80% have acquired CD45RO while maintaining high expression of CD45RA, although 10-47% are able to maintain exclusive expression of CD45RA despite their more differentiated state. In contrast, stimulation with PHA/PMA, which gives a vigorous proliferative response, appears to inhibit acquisition of CD3 and thus differentiation, while forcing a premature transition from CD45RA to CD45RO. These in vitro systems appear to permit generative thymic development while maintaining a cycling multinegative subset, thus mimicking thymic development in vivo, permitting an exploration of the events in positive and negative selection of human thymocytes. The maintenance of CD45RA expression with co-expression of CD45RO by CD3+ progeny in cultures supplemented with IL-2 or anti-CD2/28, in contrast to the loss of CD45RA after PHA stimulation, shows that the regulation of CD45 isoform expression is closely linked to the nature of the developmental signals received by a thymocyte. PMID- 8225398 TI - Production of transferrin by human cell lines in a defined protein-free system: detection by ELISA. AB - A double sandwich enzyme immunoassay using a combination of two polyclonal and one monoclonal antibodies was developed to quantitate transferrin in various human cell lines (U 937, K 562, Jiyoye, Jurkat, Raji, THP-1, HeLa) and transferrin secreted by these cells cultured in a defined protein-free medium. In the developed assay the measurable range in our experimental system is 1 to 25 ng/mL of human transferrin and the interassay coefficient of variation ranged from 11.3 to 16.1%. The assay is specific for both transferrin and apotransferrin with the same sensitivity. Intracellular transferrin levels in all the cell lines tested were < 1 ng/10(6) cells. The highest level of transferrin was found in cells of the histiocytic line U 937 (0.84 +/- 0.18 ng/10(6) cells). Only this cell line secreted transferrin in amounts > 1 ng/10(6) cells per day, i.e. 15.8 +/- 2.3 ng/10(6) cells per day. Intracellular transferrin was also demonstrated by immunoblotting. PMID- 8225399 TI - Control of clonal deletion in the thymus: implications for tolerance induction. AB - Tolerance to foreign antigen by means of clonal deletion in the thymus can be achieved during embryonic or neonatal life, but not easily in the adult animal. It is concluded here that an immunological control by mature T cells prevents newly acquired antigen from being permanently deposited in the adult thymus. In this way, the originally established thymic antigen discrimination can be maintained successfully. Breakdown of the control circuit should permit tolerance induction even in the adult thymus. PMID- 8225400 TI - Molecular cloning and characterization of a ruminant interleukin-6 cDNA. AB - By hybridization with a human interleukin-6 (IL-6) cDNA fragment a corresponding ruminant (ovine) cDNA was isolated from a lipopolysaccharide (LPS)-stimulated alveolar macrophage library. The nucleotide sequence of the cDNA and the predicted amino acid sequence of the protein showed significant homology to the human and murine molecules. Ovine IL-6 cDNA encodes a polypeptide of 208 amino acids that, based on analysis of human IL-6, is processed to a protein of 180 amino acids. Northern blot analysis and the 7TD1 bioassay were used to analyse regulatory aspects of IL-6 production by primary ovine fibroblasts. Both LPS and recombinant ovine IL-1 alpha were shown to induce IL-6 mRNA with peak levels occurring at 1 h post-stimulation and declining thereafter. When fibroblasts were pretreated with cyclohexamide prior to stimulation the level of induction by LPS and IL-1 alpha increased dramatically and peak levels were observed at 5 h post stimulation. The level of secreted IL-6 increased rapidly over the first 24 h and continued to increase over the next 48 h. PMID- 8225401 TI - Immunosuppressive antibody treatment prolongs graft survival in two murine models of segmental pancreas transplantation. AB - Successful pancreas transplantation requires the suppression of both allograft rejection and recurrence of autoimmune disease. In order to study treatments to suppress these two responses, separate models were developed for pancreas allograft rejection and autoimmune disease. In the first model, the diabetic state was induced with streptozotocin in CBA mice prior to the transplantation of pancreas grafts from BALB/c donors. In the absence of autoimmune disease, control mice rejected their grafts in 26 days (median). Antibody treatments (anti lymphocyte serum and anti-CD4) significantly prolonged allograft survival beyond this time, but not to the extent we have previously reported in the heart graft model. NOD/Lt mice spontaneously developed autoimmune diabetes, and recurrence of disease was seen in isografts at 9.5 days (median). Antibody treatments significantly delayed disease recurrence, with anti-CD4 being the most effective. Heart allografts (CBA donors) in NOD/Lt recipients were rejected within 17 days (median), and the anti-CD4 treatment had a moderate effect in delaying graft survival (median 28 days). Anti-lymphocyte serum did not prolong graft survival. Thus antibody treatment was effective in delaying both rejection and the recurrence of autoimmune disease in segmental pancreas grafts. However, the same doses were not effective in delaying heart rejection in the NOD/Lt model, so it would appear that treatments which inhibit autoimmune disease may not prevent allograft rejection. PMID- 8225402 TI - Human recombinant interleukin-1 receptor antagonist (hrIL-1RA) inhibits prostaglandin E2 (PGE2) generation but not alkaline phosphatase activity in in vivo chronic granulomatous tissue induced by KMnO4. AB - Interleukin-1, a soluble polypeptide, plays an important role in inflammatory reactions by increasing prostaglandin E2 (PGE2) generation. Human recombinant IL 1 receptor antagonist (hrIL-1ra) is a natural inhibitor of IL-1 which blocks its activity in several inflammatory states. In these studies we found that hrIL-1ra (250 mg/ml) inhibits the generation of PGE2, as measured by RIA method, in minced mouse granuloma tissue (700 mg) treated overnight with LPS (10-1000 ng/ml) or hrIL-1 beta (0.1-10 ng/ml). In addition, we show that hrIL-1ra (250 ng/ml) strongly inhibited IL-1 alpha and IL-1 beta, as measured by ELISA method, in the minced granuloma tissue treated overnight with LPS 1 micrograms/ml or IL-1 beta (10 ng/ml). The granuloma tissue induced in mice by a dorsal subcutaneous injection (0.2 ml) of a saturated solution (1:40 dilution) of KMnO4 crystals, presented an alkaline phosphatase activity which was not inhibited by two intraperitoneal administrations of hrIL-1ra 20 micrograms/200 ml bolus injections (given at the same time as KMnO4 injection and one 24 h later). These results show for the first time that hrIL-1ra blocks PGE2, IL-1 alpha and IL-1 beta but not alkaline phosphatase activity, which is a marker in growing bone and in calcific and inflamed tissue. PMID- 8225403 TI - Fungal beta-glucans modulate macrophage release of tumor necrosis factor-alpha in response to bacterial lipopolysaccharide. AB - Tumor necrosis factor-alpha (TNF alpha) is a potent cytokine believed to participate in the development of endotoxin-induced shock and the adult respiratory distress syndrome. Treatment of animals with beta-glucan prior to bacterial challenge reduces TNF alpha release and prevents death. We therefore hypothesized that beta-glucan might regulate TNF alpha secretion from macrophages in response to lipopolysaccharide (LPS). Rat alveolar macrophages were cultured in the presence of beta-glucan alone and the TNF alpha secretion quantified using an L929 cytotoxicity assay. Concentrations of beta-glucan less than 500 micrograms/ml were found to stimulate TNF alpha release from macrophages. However, concentrations of beta-glucan greater than 500 micrograms/ml resulted in suppression of the TNF alpha activity released. This reduction in TNF alpha release was not mediated by a toxic effect of beta-glucan, as large concentrations of beta-glucan had no effect on macrophage viability. We further observed that the incubation of macrophages with large concentrations of beta glucan (500 micrograms/ml) also inhibited the secretion of TNF alpha induced by bacterial LPS. Furthermore, interferon-gamma (IFN gamma), a potent activator of TNF alpha expression, failed to overcome the inhibition of TNF alpha caused by beta-glucan. These data suggest an immunomodulatory role for beta-glucan which may explain both the TNF alpha-stimulating and -inhibiting effects of fungal beta glucans during infection. PMID- 8225404 TI - Activity of human peritoneal macrophages against a human tumor: role of tumor necrosis factor-alpha, PGE2 and nitrite, in vitro studies. AB - Human peritoneal macrophages collected from renal patients on continuous ambulatory peritoneal dialysis (CAPD) during inflammation-free periods were induced to express antitumor activity in vitro when cultured in the presence of bacterial lipopolysaccharide (LPS) and even more activity when they were kept in the presence of LPS + IND (indomethacin). The antitumor activity was expressed against a human tumor-cell line, RC43, either in a cell-to-cell contact set-up between the macrophages and the RC43 target cells or when the tumor cells were exposed to supernatants of the cultured macrophages. The antitumor activity of macrophages was correlated to a marked increase in production of tumor necrosis factor-alpha (TNF alpha), not correlated to an increase in nitrite production and inversely correlated to the production of PGE2. The RC43 tumor cells were susceptible to recombinant human TNF alpha, recombinant human IL-1 beta, sodium nitrite and the leukotriene LTB4. The results obtained suggest that activated human macrophages might represent a useful tool for cancer immunotherapy. PMID- 8225405 TI - The human monocyte cell line U937 binds HIV-1 gp41 by proteins of 37, 45, 49, 62 and 92 kDa. AB - Based on our findings that HIV-1 gp41 independently of CD4 can bind to the helper T cell line H9 and B cell line Raji, we characterized the putative binding of HIV 1 gp41 to the monocyte cell lines U937 and HL60. Using flow cytometry (FACS) we examined the binding of soluble gp41 (sgp41; env amino acids 539-684) to these monocyte cell lines. Using sgp41 attached to Sepharose beads, U937 cell lysates were absorbed. The sgp41 eluate of U937 cell lysates could inhibit sgp41 binding to U937 cells. With SDS-PAGE of sgp41 eluate of U937 cell lysates, three strong protein bands, (37, 45 and 62 kDa) and two weak bands (49 and 92 kDa) were stained with Coomassie blue. With Western blot (ligand blot) analysis using sgp41, three strong protein bands (37, 49 and 62 kDa) and a very weak band (42-45 kDa) were observed in sgp41 eluate of U937 cell lysates. The results suggest that the four proteins 37, 42-45, 49 and 62 kDa in U937 cell lysates are possible candidates for the putative gp41 receptor(s). We compared the blocking activities of sgp41 eluates from different cell lysates. Not only U937 and Raji lysate-sgp41 eluates, but also H9 and HL60 lysate-sgp41 eluates could block sgp41 binding to U937 and Raji cells. The results indicate that the sgp41-binding proteins on U937, or Raji (H9 and HL60, respectively) probably could have an identical blocking (or binding) specificity; these cell types carry very similar receptor(s) for HIV-1 gp41 binding. PMID- 8225406 TI - Identification of a 16-kDa thymocyte membrane glycoprotein involved in the thymocyte/thymic medullary epithelial cell interaction. AB - We have previously described a type of lymphoepithelial interaction involving CD4+ CD8+ thymocytes and a medullary epithelial cell line (E-5). This interaction is mediated by the recently described gp23/45 epithelial adhesion molecule and an as yet unknown thymocyte receptor. The present work describes a thymocyte surface glycoprotein of 16 kDa which binds both to E-5 cells and to the purified gp23/45 adhesion molecule. In addition, a thymic lymphoma cell line (Ti-6), which interacts with the E-5 cells via the gp23/45 receptor, also present a 16-kDa glycoprotein on its surface. Taken together, the data suggest that the 16-kDa thymocyte surface glycoprotein participates in the binding between these cells and the thymic epithelium. PMID- 8225407 TI - Analysis of early reconstitution events in the SCID mouse thymus following rat bone marrow cell transplantation. AB - In this work, we provide comprehensive evidence that sublethally irradiated Thy 1.2 SCID mice can be used as a model system for thymus homing and reconstitution after intravenous transfer of rat bone marrow cells. Full short-term SCID mouse thymus reconstitutions were obtained using a plastic nonadherent low-density rat bone marrow cell subset. Cell counts and flow cytometric analysis showed that at 3 weeks post-transfer the SCID mouse thymus contained up to 41 x 10(6) Thy 1.1high rat lymphoid cells comprising the expected percentages and distribution of CD2+, CD5+, CD3+, alpha beta TCR+ and CD4+ CD8+ cells. As seen on cryostat sections, bone marrow-derived MHC class II+ accessory cells had already developed by 2 weeks post-transfer, preceding the thymocyte expansion apparent at 3 weeks. Thus, analysis of the early events of SCID thymus reconstitution by rat bone marrow cells shows that they closely resemble those described in congenic animals and points out the temporally distinct development of dendritic cells and thymocytes. The SCID mouse-rat chimera model system represents a new in vivo tool for manipulating rat T-cell differentiation from bone marrow-resident precursor cells and in addition supports our previous xenogeneic reconstitution studies performed in organ culture. PMID- 8225408 TI - Membrane expression of IL-1 alpha in chronic B lymphocytic leukemia. AB - Patients with B chronic lymphocytic leukemia (B-CLL) demonstrate a high variability in disease activity, from a benign monoclonal lymphocytosis to a rapidly fatal condition. Progressive B-CLL is related to a high expression of different growth factor receptors on the leukemic cells and to a high proliferative in vitro response to Staphylococcus aureus strain Cowan 1 (SAC). As the expression of membrane IL-1 alpha (mIL-1 alpha) indicates B lymphocyte activation, we have investigated mIL-1 alpha on cells from patients at different stages of the disease. Total cellular levels of IL-1 alpha were measured by flow cytometry of permeabilized cells and compared with CD5, CD19, CD25 and IgM expression on the cell surface. mIL-1 alpha is upregulated, both in leukemic and normal lymphocytes, in response to sIgM cross-linking with SAC or phorbol ester activation. A significantly higher expression of mIL-1 alpha was found in cells from patients with a clinically benign form of the disease. PMID- 8225409 TI - Production of interleukin 8 (IL-8) by cord blood mononuclear cells induced by Listeria monocytogenes. AB - The defective ability of human newborns to mobilize phagocytes to the site of infection led us to examine the ability of cord blood mononuclear cells to secrete interleukin-8, a major neutrophil chemotactic factor, in response to stimulation with Listeria monocytogenes. Adult or cord blood mononuclear cells were incubated with L. monocytogenes for varying lengths of time, and IL-8 was measured in the culture supernatants by the enzyme-linked immunosorbent assay (ELISA). Spontaneous IL-8 secretion by unstimulated cells was undetectable or at the minimal detection limit of the assay. By 24 h of cell incubation with L. monocytogenes, newborn cells produced as much IL-8 as adult cells did (300 +/- 113 versus 269 +/- 189 ng/ml, respectively). Over the next 2-4 days, IL-8 output by adult cells was slightly higher than that by newborn cells, but the difference was not statistically significant. The in vitro results suggested that newborns are as able as adults to produce IL-8, although they are defective in mobilizing neutrophils, the IL-8 target cells, to the site of infection. PMID- 8225410 TI - The relative density of CD44-positive porcine monocytic cell populations varies between isolations and upon culture and influences susceptibility to infection by African swine fever virus. AB - African swine fever (ASF) virus has been reported to infect cells of the monocyte family, probably macrophage-like cells, but there is variation in the apparent susceptibility of these cells. We have demonstrated that the phenotype and activity of porcine monocytic cells varies between different isolations and also upon culture. The variation during culture is dependent upon the phenotype of the cells at the time of isolation. As for the susceptibility of porcine monocytes/macrophages to infection by ASF virus, it was seen that this could be related to the variation in cell phenotype and activity. The susceptibility was determined by the relative density of particular subpopulations of cells present. Whilst inflammatory macrophages did not have an apparent role to play, phagocytic activity was influential. Furthermore, the expression of CD44 and the DH59 myeloid cell marker was important, whereas the relevance of MHC Class II expression was variable. Overall, it was concluded that susceptibility to infection required that a culture be dominated by CD44-positive cells which were non-inflammatory, of low phagocytic activity, and characterizable as being of the myeloid (DH59-positive) lineage. PMID- 8225411 TI - IL-2 and IL-3 production in high and low IgG-responding strains of mice. AB - The antibody response of mouse strain C57Bl/10ScSn (B10) is characterized by a low IgG responsiveness to a number of different antigens. Aberrant function of antigen-presenting cells and/or low activity of the Th cell population have been suggested as the cause of the defect. We studied the production of IL-2 and IL-3 in vitro by unstimulated and ConA-stimulated spleen cells. Unstimulated spleen cells of low-responding B10 mice produce significantly less IL-2 compared with the high-responding A/J mice in both intervals tested, i.e. after 24 and 48 h of in vitro incubation. IL-3 production is low but almost comparable in unstimulated cells of both strains. Stimulation of spleen cells by 5 micrograms/ml of ConA leads to considerably higher production of IL-2 in A/J spleen cells. IL-3 production by ConA-stimulated spleen cells showed the same pattern of activity. This low IL-3 production by B10 cells is most likely due to the low production of IL-2 during Th cell activation and to the limited proliferation of these cells. The low IgG production of B10 spleen cells during the secondary response to SRBC in vitro could be restored by IL-2 added to the medium. 50 U/ml of IL-2 increased the number of anti-SRBC IgG-producing cells 40 times in B10 cells, but only 4 times in A/J cells, so that the IgG production in B10 cells reached the same level as that in the A/J cells without exogenous IL-2. We suppose that the limited IL-2 production in the low-responding strain B10 is the cause of the low IgG responsiveness of these mice. PMID- 8225412 TI - Generation and characterization of monoclonal antibodies to porcine zona pellucida-3 alpha glycoprotein. AB - All 10 monoclonal antibodies generated reacted with deglycosylated ZP3 alpha both in ELISA and Western blots, thereby suggesting that these do not recognize carbohydrate determinants. Moreover, 7 antibodies (MAs -402, -403, -405, -412, 420, -421 and -423) recognized reduced and carboxyamidomethylated ZP3 alpha in Western blot suggesting that these antibodies recognized linear epitopes. The epitopes recognized by MAs -410, -413 and -425 were sequential or conformational, stabilized by disulphide bonds. Three antibodies namely, MA -405, -420 and -421 inhibited in vitro, zona lysis by trypsin. PMID- 8225413 TI - Specific activities of seminal vesicular phosphomonoesterases and Mg2+-,Ca2+- and Na+/K(+)-adenosine triphosphatases in hypo- and hyperthyroid albino rats. AB - Hypothyroidism (surgical thyroidectomy) inhibited the activities of acid phosphatase and Mg(2+)-ATPase in seminal vesicular tissue and fluid and that of Ca(2+)- and Na+/K(+)-ATPases in fluid alone, and T4 supplementation restored normalcy in all, except acid phosphatase. Hyperthyroidism (T4 25 micrograms/100g body weight/day for 60 days, im) enhanced the activities of alkaline phosphatase and ATPases in seminal vesicular tissue and fluid, and decreased acid phosphatase activity in tissue alone. Withdrawal of T4 treatment from hyperthyroid rats (after 30 days) augmented the activity of ATPases in tissue and impaired the same in fluid, while phosphomonoesterases remained at hyperthyroid level. The results suggest specific responses of various seminal vesicular phosphatases to altered thyroid hormone status. Modification in the specific threshold of androgen/estrogen action on different phosphatases in seminal vesicles appears to be the plausible mechanism underlying these changes in hypo- and hyperthyroid conditions. PMID- 8225414 TI - Growth response of Pseudomonas stutzeri RS34 to ethylenediaminetetraacetic acid (EDTA) and its interaction with zinc. AB - Pseudomonas stutzeri RS34 is a less sensitive member of pseudomonads to toxic effect of EDTA, the effect of EDTA is more bacteristatic than bactericidal, and can be reversed by divalent cations. Zn2+ provides more specific protection than Mg2+. EDTA-treated cells show higher sensitivity to lysozyme confirming the chelating mode of action of EDTA that leads to destabilization of the outer membrane. Such metal resistant bacteria can be profitably employed in the removal of metals from polluted ecosystems. PMID- 8225415 TI - Effect of indigenous drugs on glucuronoglycan metabolism in diabetic hypertensive rabbits. AB - Development of hypertension further exaggerated the deranged level of total glucuronoglycan (GAG), different fractions of GAG, activities of synthetic and degradative enzymes of GAG and biological sulphation observed in diabetic rabbits. Administration of indigenous drug preparations (nisakathakathi Kashayam and rasnairandadi Kashayam and its mixture) ameliorated the diabetic hypertensive condition by reversing these effects and thereby arrested. PMID- 8225416 TI - Role of media constituents and proline in callus growth, somatic embryogenesis and regeneration of Oryza sativa cv Indica. AB - Conditions were optimised for efficient callus induction from seeds of four local indica rice cultivars, GR-3, GR-102, Jaya and Te-Tep. Addition of 2,4-D to MS medium at 2.5 mg/l resulted in 100% callus induction. N6 medium was superior to MS medium for callus growth, formation of embryogenic callus as well as regeneration. Cultivar Te-Tep showed highest callus growth while GR-102 the least. Addition of casein hydrolysate enhanced growth of callus but did not yield more embryogenic calli. Supplementation of MS or N6 media with proline, not only increased callus growth but also showed an increase in embryogenic callus formation. GR-102 callus was most embryogenic followed by Te-Tep, GR-3 and Jaya. Histological observation of embryogenic calli revealed the presence of pro-embryo like structures. It was also observed that calli induced on N6 medium supplemented with proline could maintain regeneration potential for a longer period as compared to other media. Cytokinins like BAP or kinetin alone could not initiate shoot formation. Regeneration frequency, and the number of shoots formed per callus increased significantly. Cultivar Te-Tep gave the best response for regeneration followed by Jaya, GR-3 and GR-102. PMID- 8225417 TI - Antifilarial effect of two triterpenoid saponins isolated from Acacia auriculiformis. AB - The active principle, isolated from the funicles of A. auriculiformis, consisted of two triterpenoid saponins, acaciaside A and acaciaside B which killed in vitro 97% microfilaria of Setaria cervi in 100 min at 4mg/ml concentration and 100% of adults in 35 min. The drug, when administered orally at 100 mg/kg on rats, in which S. cervi adults were implanted intra-peritoneally, increased the blood mf count by 1.5-fold after the first phase of treatment for 10 days. Following the third phase of treatment and thereafter, the mf density was reduced by more than 80%. No toxic effect of the saponins was observed in rats. The rise in mf count indicated that the drug induced a very high physiological stress on the adult worms which increased the rate of discharge of the mf before impending death. The treated rats on autopsy did not show any adult worms. PMID- 8225418 TI - Antitumor and radiosensitizing effects of Withania somnifera (Ashwagandha) on a transplantable mouse tumor, Sarcoma-180. AB - Antitumor and radiosensitizing effects of alcoholic root extract of W. somnifera and their modification by heat were studied in vivo on Sarcoma-180 grown on the dorsum of adult BALB/c mouse. Ashwagandha (AT) was injected (ip) at a dose of 500 mg/kg body wt for 10 consecutive days into mouse bearing tumor of 50 +/- 5 mm3, with or without a local treatment of 10 Gy gamma radiation (RT) or hyperthermia at 43 degrees C for 30 min (HT) or both on 5th day of AT. The response was assessed on the basis of tumor regression, growth delay, animal survival and changes in the tumor GSH content. Ashwagandha, RT and HT individually produced 18, 38 and 45% complete response (CR) respectively, but RT gave the best long term survival. Ashwagandha increased the effect of radiation on tumor regression as well as growth delay, but AT + HT gave a better tumor cure. However, both these combinations gave almost identical long term survival, which was not much higher than that produced by RT alone. The combination of Ashwagandha for 10 days with one local exposure to RT followed by HT significantly increased the tumor cure, growth delay of partially responding tumors and animal survival. This combination also significantly and synergistically depleted the tumor GSH level, with no recovery even at 3 hr after treatment. It is concluded that Ashwagandha, in addition to having a tumor inhibitory effect, also acts as a radiosensitizer and heat enhances these effects. The severe depletion in the tumor GSH content by the combination treatment must have enhanced the tumor response, as the inherent protection by the thiol will be highly reduced. PMID- 8225419 TI - Loss of electrolytes during repair and regeneration of skeletal muscle. AB - Changes in the specific conductivity and electrolytes have been studied in the leachates from normal and minced autografted chick gastrocnemii with special reference to the loss of Na+, K+, Ca2+, Mg2+ and PO(4)3- ions. The study has revealed age related modifications in the muscle membranes as well as the presence of some intrinsic regulatory mechanisms which get stimulated under the induced-stress condition. Higher ionic efflux in the regenerating muscle indicates that the anatomical regeneration does not correspond to functional regeneration of muscle membranes. PMID- 8225420 TI - Angiotensin converting enzyme activity in female reproductive tract of domestic fowl (Gallus domesticus) and its interactions with semen. AB - Presence of angiotensin converting enzyme (ACE, EC 3.4.15.1) in female reproductive tract and its interaction with semen has been investigated in domestic fowl. Higher activity of ACE was detected in homogenates of infundibulum region followed by magnum and relatively low level was noticed in rest of the parts of the oviduct (i.e. isthmus, uterus, uterovaginal-junction and vagina). This decreasing order in ACE activity was, however, reversed when these segments were incubated with semen. ACE in the ovarian follicular wall increased with corresponding increase in the follicle size. Presence of some stimulatory or inhibitory substances for ACE in the oviduct of fowl is suggested. PMID- 8225421 TI - Effect of intensity and duration of stress on male sexual behaviour. AB - Male rats were exposed to four different stress conditions, varying in duration and intensity, during separate periods and their copulatory behaviour was studied. The number of mounts and time spent in other sexual behaviours were reduced with intensity of stressor being primarily responsible rather than prolongation of duration of a less severe stress condition. PMID- 8225422 TI - Endothelin-1 and okadaic acid induced contractions in tracheal smooth muscle of lamb. AB - Stimulation of lamb tracheal smooth muscle fibre strips with endothelin-1 and okadaic acid results in the development of isometric tensions which are long lasting. However, endothelin-1 is more potent constrictor than okadaic acid. An analysis of 20,000 Da regulatory myosin light chain by two-dimensional gel electrophoresis shows an identical phosphorylation pattern. PMID- 8225423 TI - New microbiological method for the detection of staphylococcal beta-lactamase. AB - The method has been designed as such to include the induction and testing of beta lactamase in a single procedure. This method compares favourably with the existing method. PMID- 8225424 TI - In vitro absorption of nutrients from small intestine of rats fed peroxidized oil. AB - Feeding of diet containing peroxidized oil (peroxide value approximately 90 mEq/kg) at 10% level for 4 weeks altered the absorptive function of intestine. Rate of in vitro absorption of nutrients especially sucrose and protein hydrolysate (amino acids) was considerably reduced. Absorption of glucose, fructose and Ca2+ was only marginally affected. PMID- 8225425 TI - Is the X-chromosome early replicating in mitotic cells of Drosophila males? PMID- 8225426 TI - Asthma in the paediatric age-group. PMID- 8225427 TI - Postoperative pulmonary complications and serial pulmonary functions and arterial blood gases in patients undergoing oesophagectomy with or without a thoracotomy. AB - Transthoracic (TT) (n = 12) and transhiatal oesophagectomy (THE) (n = 14) were performed in 26 patients (19 males and 7 females) suffering from benign or malignant strictures of oesophagus. Nine of these patients were smokers. Preoperatively, various parameters of pulmonary functions did not differ significantly in two groups of patients. Forced vital capacity (FVC), forced expiratory volume in 1 second FEV1 (L) and forced expiratory flow between 25-75% of vital capacity (FEF 25-75%) showed a decline at second/third, seventh and 6 weeks in the postoperative period compared to preoperative values. Decrease in FVC was significant at all times. The change in FEV1 and FEF25-75% was not statistically significant at 6 weeks. The decrease in arterial oxygen tension (PaO2), maximal breathing capacity (MBC), maximal inspiratory (PImax) and expiratory (PEmax) pressures in the postoperative period was significant at second/third and seventh postoperative days. Three of 26 patients died. Of these two had TT while the third one had THE procedure. Twenty-one patients developed postoperative pulmonary complications (PPCs). Patients who developed PPCs had significantly different MBC, FEF25-75%, PEmax and FEV1 compared to those who did not have PPCs. Long-term studies involving a larger number of patients are required to document changes occurring in pulmonary functions, arterial blood gases and nutritional status of these patients. PMID- 8225428 TI - Role of fine needle aspiration cytology in the diagnosis of lymphadenopathy. AB - Fine needle aspiration cytology (FNAC) and open biopsy of same enlarged lymph nodes were performed in 59 patients with lymphadenopathy. When histopathological and cytological examinations were compared, an accuracy of 91.5% was achieved with FNAC in the cases of lymphadenopathy of various etiologies. Thus, fine needle aspiration cytology being reliable, safe, rapid and economical procedure, is useful as an outdoor diagnostic procedure for diagnosing lymphadenopathy of various etiologies. PMID- 8225429 TI - Effect of routine fiberoptic bronchoscopy and bronchoalveolar lavage on arterial blood gases. AB - Effect of conventional fiberoptic bronchoscopy (FOB) procedure (Group A, n = 10) and bronchoalveolar lavage (BAL) (Group B, n = 11) on arterial blood gases was studied in 21 patients. A significant fall in arterial oxygen tension (PaO2) with widening of alveolar arterial oxygen gradient D(A-a)O2 was observed during FOB (p < 0.001) and 30 minutes following the procedure (p < 0.001). Bronchoalveolar lavage was also associated with significant fall (p < 0.001) in PaO2 with increased D(A-a)O2 and this fall in PaO2 persisted for 30 minutes after the procedure (p < 0.01). None of the patients received oxygen during the procedures. None of the patients in either group developed bronchospasm. No change in arterial carbon dioxide (PaCO2) was observed in either group. Hypoxaemic encephalopathy developed in one patient in group A, hypotension and cyanosis developed in one patient each in group B. No mortality was encountered in the study. All the three patients with complications were excluded from final analysis. It was concluded from this study that both conventional bronchoscopy and BAL procedures were associated with significant hypoxaemia and supplemental oxygen should be given to all patients during the procedure and should be continued for at least 30 min. afterwards. PMID- 8225430 TI - Effect of steam inhalation on mucociliary activity in patients of chronic pulmonary disease. AB - Increased mucociliary activity has been observed with several modalities, including inhalation which seems to achieve faster drug delivery. In patients of chronic lung disease, the mucociliary activity is known to be impaired. The effect of steam inhalation on mucociliary transport time was studied in these patients. It was found that steam inhalation improved mucociliary activity significantly (p < 0.001) in both groups receiving either only bronchodilators, or bronchodilators as well as steroids. PMID- 8225431 TI - Idiopathic pulmonary haemosiderosis: an unusual cause of recurrent haemoptysis. AB - Haemoptysis is a clinical problem which occurs in association with a variety of disorders. We report a patient who presented with the triad of haemoptysis, anaemia and diffuse alveolar consolidation suggesting diffuse pulmonary haemorrhage (DPH). PMID- 8225432 TI - Macleod's syndrome. PMID- 8225433 TI - Bilateral spontaneous pneumothorax in silicosis. AB - Occurrence of pneumothorax in silicosis is rare. We report here one case of silicosis who presented with bilateral spontaneous pneumothoraces occurring simultaneously. PMID- 8225434 TI - Prevalence of asthma and wheeze in two different climatic areas of Saudi Arabia. AB - In a cross-sectional study, the prevalence of asthma and wheeze was studied in 2300 school going children in two different environment and climate regions of Saudi Arabia, Dammam (humid-coastal) and Riyadh (dry-inland). The age range of children studied was 7-12 years (56% boys and 44% girls in Dammam; and 47% boys and 53% girls in Riyadh). A self-administered questionnaire was completed by parents of the children which includes information on age, sex, area, occupation and educational level of parents, history of asthma, allergic rhinitis, eczema, cough, wheeze, animals and pets exposure, family history of allergy and parental smoking habits. The results showed that prevalence of wheeze was more common in Riyadh (11.86%) than in Dammam (6.54%). Also, cough occurred more frequently in Riyadh (7.9%) than in Dammam (6%). The frequency of rhinitis was significantly more common in both areas than wheeze (Riyadh 17% and Dammam 12%). Also, the attack of breathlessness or tightness occurred more frequently in Riyadh (12.13%) than in Dammam (6.10%). When a more formal diagnosis of asthma was sought, this having been made by a doctor, the figures were 9.28% for Riyadh and 3.59% for Dammam. These figures showed major differences between the coastal and inland areas in the prevalence of bronchial asthma and wheezing with Dammam being relatively low risk whereas children in Riyadh region describe this more commonly. PMID- 8225435 TI - Distinct genetic pattern of mouse susceptibility to thyroiditis induced by a novel thyroglobulin peptide. AB - Experimental autoimmune thyroiditis (EAT), induced by thyroglobulin (Tg) and adjuvant, is major histocompatibility complex-controlled and dependent on Tg reactive T cells, but the immunopathogenic T-cell epitopes on Tg remain mostly undefined. We report here the thyroiditogenicity of a novel rat Tg peptide (TgP2; corresponding to human Tg amino acids 2695-2713), identified by algorithms as a site of putative T-cell epitope(s). TgP2 causes EAT in SJL (H-2s) but not in C3H or B10.BR (H-2k), BALB/c (H-2d), and B10 (H-2b) mice. This reveals a new genetic pattern of EAT susceptibility, since H-2k mice are known to be high responders (susceptible) after Tg challenge. Following in vivo priming with TgP2, T cells from only SJL mice proliferated significantly and consistently to TgP2 in vitro, whereas TgP2-specific IgG was observed in all strains tested. Adoptive transfer of TgP2-primed SJL lymph node cells to naive syngeneic recipients induced a pronounced mononuclear infiltration of the thyroid, which was more extensive than that observed after direct peptide challenge. TgP2 is non-immunodominant, since priming of SJL mice with rTg did not consistently elicit T-cell responses to TgP2 in vitro and a TgP2-specific T-cell hybridoma did not respond to antigen presenting cells pulsed with rTg. The data support the notion that Tg epitopes need not be either iodinated or immunodominant in order to cause severe thyroiditis and that the genetic pattern of the disease they induce can be distinct from that of Tg-mediated EAT. PMID- 8225436 TI - Susceptibility to immunosuppression by ultraviolet B radiation in the mouse. AB - Irradiation with ultraviolet B (UVB; 290-320 nm) initiates systemic immunosuppression of contact hypersensitivity (CHS). UV dose-responses for suppression of CHS to trinitrochlorobenzene were established in 18 strains of inbred mice. Three phenotypes with significantly different susceptibilities to UV suppression were identified. The phenotypes were: high (HI) susceptibility, 50% suppression with 0.7-2.3 kJ/m2 UV (C57BL/6, C57BL/10, and C57L and NZB females); low (LO) susceptibility, 50% suppression with 9.6-12.3 kJ/m2 UV (BALB/c, AKR, SJL and NZW), and intermediate (INT) susceptibility, 50% suppression with 4.7-6.9 kJ/m2 UV (DBA/2, C57BR, C3H/HeJ, C3H/HeN, CBA/N and A/J). UV suppression was not correlated with skin pigmentation or with the magnitude of the CHS response in non-irradiated animals. Major histocompatibility complex (MHC) haplotype was not correlated with UV suppression in MHC congenic strains B10.D2/oSnJ, B10.D2/nSnJ, B10.BR/SgSnJ, and A.BY/SnJ. There were no sex differences in UV suppression in BALB/c, C57BL/6, or NZW animals. In the autoimmune NZB strain, however, male mice (LO) were seven times less sensitive to UV suppression than NZB female mice (HI). Both sexes of (NZB x NZW)F1 and (NZW x NZB)F1 mice were HI, supporting dominance of HI over LO. Thus there are genetic factors and interacting sex-limited factors determining susceptibility to UV suppression. These findings may be of relevance to UV-related diseases such as photosensitive lupus and skin cancer. PMID- 8225437 TI - Molecular analysis of an HLA-DP mutant cell line selected for its resistance to killing by HLA-DPw2-specific T-cell clones. AB - A collection of HLA-DP mutants was generated, using ICR 191 as the mutagenic agent and resistance to lysis mediated by HLA-DPw2 allospecific cytotoxic T lymphocytes (CTLs) as the selection criterion. These mutants were derived from the HLA haploid lymphoblastoid cell line 45.1. Loss of HLA-DPw2 surface expression accounted for the lack of HLA-DPw2 CTL recognition in all the mutants. However, one of them, 45.EM19, binds to DPw2-specific monoclonal antibodies (mAb) after cell permeabilization. HLA-DPA1 and DPB1 mRNA expression studies permitted the classification of the mutants in four categories: 1) DPA1-negative, DPB1 positive; 2) DPA1-positive, DPB1-negative; 3) DPA1- and DPB1-negative, and 4) DPA1- and DPB1-positive mutants. Mutant 45.EM19 is included in the last group. The cloning and sequencing of the full-length DPA1 (DPA1*0103) and DPB1 (DPB1*02012) cDNAs from this mutant showed no changes in the DPA1 sequence compared to the wild-type sequence. However, a frame-shift mutation in the DPB1 gene exon coding for the transmembrane region was detected. The insertion of a guanine nucleotide provokes an extension of the open reading frame, increasing the length of the C-terminal domain and changing the hydropathicity pattern of the transmembrane domain. This change should be responsible for the phenotype of the 45.EM19 mutant. PMID- 8225438 TI - Limited polymorphism in HLA-DM does not involve the peptide binding groove. PMID- 8225439 TI - HLA class I/class II interaction in early onset pauciarticular juvenile chronic arthritis. PMID- 8225440 TI - Coding sequence polymorphism of Tcrg-V1, -V2, and -V4 genes in mice bearing Tcr gA and -gC haplotypes. PMID- 8225441 TI - Dominant aromatic/aliphatic C-terminal anchor in HLA-B*2702 and B*2705 peptide motifs. PMID- 8225442 TI - Nucleotide sequence of a novel HLA-DRB1 allele, DRB1*0810 [corrected]. PMID- 8225443 TI - A novel HLA-C allele (Cw*1504) related to the Cw6.2 phenotype. PMID- 8225444 TI - Nucleotide sequence of the Lewis rat granulocyte-macrophage colony stimulating factor. PMID- 8225445 TI - 34th Annual Conference of the Indian Society of Gastroenterology. Goa, October 14 17, 1993. Abstracts. PMID- 8225446 TI - Adrenal function & the pattern of glucocorticoid induced eosinopenia in tropical pulmonary eosinophilia. AB - Plasma cortisol levels were estimated in 16 patients with tropical pulmonary eosinophilia (TPE; group A) by radioimmunoassay using 125I labelled cortisol. The cortisol levels showed diurnal variation, the mean levels were 10.22 +/- 4.5 micrograms/dl for the morning samples and 4.91 +/- 2 micrograms/dl for the night samples respectively and were within normal limits. Another six patients with TPE included in the study (group B) had an initial mean plasma cortisol level of 5.83 +/- 1.33 micrograms/dl. Half an hour after intramuscular administration of 250 micrograms of Synacthen (a synthetic glucocorticoid with corticotrophic activity), the mean cortisol level showed a significant rise up to 14.7 +/- 3.54 micrograms/dl (P < 0.01) thus indicating normal adrenocorticohypophysial functions in TPE. There was no significant alteration of total blood eosinophil counts before and after intramuscular Synacthen. An additional 6 patients with TPE (group C) who were given oral prednisolone 30 mg a day for 5 days which was subsequently tapered off in the next 5 days showed a spectacular mid-treatment fall in blood eosinophil levels with a post-treatment rise. In another 3 patients with TPE (group D) total blood eosinophil counts were estimated 4 h after intramuscular hydrocortisone and a prompt eosinopenic response was observed. This indicated that a dose related response to steroids in TPE is unlikely. The absence of eosinopenic response after Synacthen could have been due to the short (half an hour) interval when the eosinophil counts were repeated. PMID- 8225447 TI - Screening for genital chlamydial infection in symptomatic women. AB - During a two year period, the prevalence of genital chlamydial infection was assessed by Papanicolaou and Giemsa cytology of endocervical specimens from 396 women attending a gynaecology outpatient department in a Delhi hospital, with symptoms of lower genital tract infection and primary or secondary infertility. The results of cytological examination were confirmed by isolation in a cell culture system. Prevalence was found to be 41 per cent (93 of 227) in patients presenting with vaginal discharge and 36 per cent (61 of 169) in infertility women. Genital chlamydial infection is prevalent in gynaecological patients in this region. PMID- 8225448 TI - A case control study of selenium in cancer. AB - As selenium has been shown to be a risk modifier of cancer at several sites in a case-control approach, selenium nutriture was studied in oral and oesophageal cancer patients. The mean selenium level in oral cancer patients was 102.6 ng/ml and in oesophageal cancer 103.3 ng/ml, while in the matched controls it was 117.5 and 116.2 ng/ml respectively. Relative risk estimates were 3.16 (P < 0.02) and 16.0 (P < 0.01) in oral and oesophageal cancer patients respectively. The risk appears to increase with selenium levels lower than 115 ng/ml. PMID- 8225449 TI - Differential patterns of malnutrition among kindergarten children in Pune city. AB - In view of the fact that inadequate nutrition is not the sole cause of existing malnutrition but is a complex interaction of social, cultural and environmental factors children growing in different social economical set-ups were studied for the differentials in types of malnutrition. Total of 300 kindergarten children aged 2-5 yr from three socio-economic classes in Pune city were examined. It was observed that even in middle socio-economic class, considerable wasting (38%) was prevalent, while stunting (86%) was the major problem of low socio-economic class children. Age-wise prevalence not only indicated high risk of malnutrition around the age of 2 yr, but also suggested tendency of temporary catch up which did not last long. As wasting and stunting have important-implications for intervention programmes, it is necessary to understand the etiology of different types of malnutrition. PMID- 8225450 TI - Effect of feeding high fat, high fiber diet on brush border enzymes in mice intestine. AB - The effect of dietary fat content on brush border enzymes has been studied in mice intestine. The results obtained from 26 per cent fat (high fat; HF)-fed mice were compared with those fed 10 per cent fat (pair-fed; PF and ad libitum-fed). Brush border alkaline phosphatase (AP), leucineaminopeptidase (LAP) and gamma glutamyltranspeptidase (gamma-GTP) activities were significantly enhanced while sucrase activity was reduced (P < 0.001) in HF group compared to the controls. Activities of lactase, p-nitrophenyl (PNP)-beta-D-glucosidase and PNP-beta-D galactosidase were unaltered under these conditions. Kinetic studies with AP, sucrase and LAP revealed that changes in enzyme levels in response to HF diet were due to change in Vmax. Significant changes in enzyme activities as a consequence of HF intake were observed in enterocytes all along the crypt-villus unit as compared to the control group. These results indicated that feeding a fat rich diet produced selective changes in brush border enzyme activities in mice intestine. PMID- 8225451 TI - Dermatoglyphic patterns of patients with rheumatoid arthritis. AB - Finger tip and palmar dermatoglyphics were studied in 31 patients (22 females and 9 males) with rheumatoid arthritis (RA) and 38 matched controls (20 females and 18 males) from North India. While not many differences were observed in palmar patterns, a low ending of line A was found on both hands of two patients. Finger tip patterns were significantly different in patients compared to controls. No association with any dermatoglyphic feature and HLA antigens was observed. PMID- 8225452 TI - Serum protein, ascorbic acid & iron & tissue collagen in oral submucous fibrosis- a preliminary study. AB - A study of 36 patients with oral submucous fibrosis, revealed that all patients had the habit of chewing betel nut, pan masala or the traditional mixture (betel nut, betel leaf and lime) suggesting a link between fibrosis and arecanut. There was an increase in the globulin fraction of protein and hence a decreased A/G ratio in these patients. There was a significant increase in total protein levels possibly due to the increase in globulin fractions and other serum proteins. Ascorbate and iron levels decreased perhaps because of their utilisation in collagen synthesis. The total tissue collagen content increased significantly in patients with advanced disease and, it increased with the progression of the disease leading to hypomobility of the tongue, lips, cheeks, soft palate and faucial pillars. PMID- 8225453 TI - Serum, CSF, RBC & urinary levels of magnesium & calcium in idiopathic generalised tonic clonic seizures. AB - Serum, cerebrospinal fluid (CSF) and urinary levels of magnesium and calcium and RBC magnesium levels were studied in 100 patients of idiopathic generalised tonic clonic seizures and 95 healthy controls matched for age and sex. There was a significant reduction in serum, CSF and RBC magnesium levels and a rise in serum and CSF calcium levels in epileptic patients. The 24 h urinary excretion of calcium and magnesium in the epileptics did not differ from controls. Post ictal (within 24 h of seizure) serum and CSF magnesium levels were significantly lower and calcium levels significantly higher as compared to inter ictal levels (4 wk after seizure). There was no correlation between serum magnesium, serum calcium and CSF calcium levels and the frequency, control or duration of fits. Low CSF magnesium levels correlated with increased frequency, poor control and longer duration of fits. Patients with status epilepticus and those in the EEG abnormalities had low CSF magnesium levels. PMID- 8225454 TI - The significance of serum magnesium in diabetes mellitus. AB - The mean serum magnesium level in normal individual was found 1.967 +/- mgm/dl. No significant difference in serum magnesium level was found in various age groups. All diabetic patients, having normal renal function, exhibited hypomagnesemia. The hyperglycemia in these cases was inversely related to hypomagnesemia and its restoration towards normal by insulin therapy restored the normal serum magnesium concentration. These existed in inverse correlationship between serum magnesium and cholesterol (r = -0.56). The hypomagnesemia was the result and not the cause of alterations in the cholesterol metabolism. A positive correlation was observed between blood urea level and serum magnesium (r = +0.7) and it was significant. The magnesium correlated with major diabetic complications too, e.g. micro and macroangiopathies. Thus, serum magnesium can be used for prognostic assessment in diabetic individuals. PMID- 8225455 TI - Clinical study of 100 cases of Kyasanur Forest disease with clinicopathological correlation. AB - 100 cases of Kyasanur Forest Disease were studied clinically and autopsy or postmortem biopsies of various organs were done in 8 cases. 1. Our study showed that KFD passes through 4 stages each lasting for about a week. i.e. a prodromal stage with fever, hypotension hepatomegaly, a stage of complication characterized by haemorrhage, neurological manifestation or bronchopneumonia, a stage of recovery followed by a li stage of fever in some cases. 2. The exact of cause of haemorrhage could not be identified though Disseminated Intravascular Coagulation was suspected. 3. Hypotension in KFD could be of Myocardial origin. 4. Encephalopathy in KFD could be due to a metabolic cause probably of hepatic origin. 5. Lung signs could be due to intraalveolar haemorrhage and secondary infection. PMID- 8225456 TI - Enteromesenteric bridge in the treatment o primary lymphedema--a case report. AB - Technical details and associated problems of enteromesenteric bridging operation for primary lymphedema (proximal obstructive hypoplasia) are discussed. The short term results are encouraging. PMID- 8225457 TI - Folate therapy in acquired lactose intolerance diarrhoea. PMID- 8225458 TI - The use of osseointegrated implants in growing jaws. PMID- 8225459 TI - Five-year statistical and clinical observations with the IMZ two-stage osteointegrated implant system. AB - This study evaluated patients who were reconstructed with the IMZ system, which consists of a cylindrical implant with an intramobile element for stress relief. It is placed through a two-stage surgical procedure resulting in osteointegration. During a 5-year period, 1,059 implants were placed in 322 patients. Twenty-one implants were lost to follow-up. A total of 28 implants failed over the 5-year period, of which 9 had not integrated at stage 2 surgery. Of the remaining implants, 19 failed primarily for periodontal and prosthetic reasons. The life table method was used for statistical analysis. The 5-year survival rate of all IMZ implants was 95%. Seventy-five percent of the implants were placed to restore partial edentulism, with a survival rate of 96%. The survival rate for totally edentulous patients was also 96%. Forty-four percent of the implants were placed in the maxilla, with a survival rate of 92%. The survival rate in the mandible was 99%. Sixty-six percent of the implants were placed in women, with a survival rate of 94%. The survival rate in men was 98%. Major factors that positively influenced long-term survival were use of the longest and largest-diameter implants appropriate for the clinical situation. PMID- 8225460 TI - Forces on implants supporting overdentures: a preliminary study of morphologic and cephalometric considerations. AB - It is usually suggested that loading forces be transmitted to implants in the vertical direction. In 44 subjects treated with mandibular implants and overdentures, the inclination of the implant axis relative to the occlusal plane of the corresponding prostheses was measured by means of cephalometric radiographs. The degree of mandibular atrophy was also assessed. Peri-implant parameters such as plaque index, bleeding scores, and probing depths were registered on four implant sites. About 70% of the implants showed a buccal inclination, 11% showed a lingual inclination, and only 19% were found with an angle of 90 degrees. After an observation period of 5 years, no positive relationship between inclination of the implant axis, mandibular atrophy, and peri-implant findings could be established. Aspects of facial morphology and the interalveolar relationship of both jaws should be considered additional parameters and should be included in investigations concerning functional dynamics of removable dentures. PMID- 8225461 TI - Binding of murine osteoblastic cells to titanium disks and collagen I gels: implications for alternative interpretations of osseointegration. AB - Using 3H-thymidine-labeled MC3T3-El osteoblastic cells, the number of osteoblasts bound to titanium surfaces after various surface treatments and incubation periods was directly measured. MC3T3-El cell binding to titanium surfaces was saturable at a low level (approximately 10,000 cells/cm2). Although treatment of these surfaces with fibronectin, keratin sulfate, and the fibronectin-derived peptide GRGDS (glycine-arginine-glycine-glutamate-serine) increases cellular binding by 29% to 31%, the relative binding to titanium was 5 to 10 times lower than binding to collagen I gels. A collagen I matrix competed with the commercially pure titanium surfaces for cell binding from solution, suggesting that direct binding of osteoblasts to titanium surfaces present within an organic matrix may not be favored. The significance of immediate and direct bone cell attachment to titanium surfaces for osseointegration should be reevaluated. PMID- 8225462 TI - The peri-implant sulcus. AB - A review of the peri-implant sulcus, including the histology of the sulcular epithelium, epithelial attachment, and the gingival connective tissue, is presented. The peri-implant junctional epithelial attachment is mediated by hemidesmosomes, as in the periodontal tissue. There is some controversy on the possible attachment of connective tissue fibers to the implant, but current studies indicate a parallel orientation with no insertion of the peri-implant connective tissue fibers. This difference in connective tissue attachment may affect the peri-implant tissue's susceptibility to disease. PMID- 8225463 TI - Treatment of ligature-induced peri-implantitis using guided tissue regeneration: a clinical and histologic study in the beagle dog. AB - This study evaluated the treatment of ligature-induced peri-implantitis using guided tissue regeneration (GTR) around submerged and nonsubmerged implants in beagle dogs. Two titanium implants were placed bilaterally in the premolar regions of the mandible in each of 10 beagle dogs. Cotton floss ligatures were placed around the implant necks and no oral hygiene procedures were performed for 5 months. Periodontal surgery was performed, and in five dogs (submerged group) the abutments were removed and an e-PTFE barrier was placed to cover the test implants and their adjacent osseous defects, while the control implants received no barriers. In the other five dogs (nonsubmerged group), flap surgery was performed without removing the abutments. In the test sites, an e-PTFE barrier was placed around the abutment and adjacent defects, while on the control sites no barrier was placed. Nondecalcified specimens were prepared using standard histologic methods and analyzed. No differences were found between any of the clinical parameters in both the control and experimental sites from the submerged and nonsubmerged groups. Histologic and histomorphometric analyses also revealed no significant differences between groups with regard to new bone formation. In the experimental (GTR) groups, however, the amount of regenerated connective tissue was significantly greater than on the control groups. Under the conditions of this study, the treatment of peri-implantitis using GTR did not enhance clinical parameters or bone formation around "diseased" implants. PMID- 8225464 TI - Observations of the anterior loop of the inferior alveolar canal. AB - The inferior alveolar nerve may extend beyond the mental foramen as an intraosseous anterior loop. The purpose of this study was to determine if panoramic radiography accurately identifies the anterior loop of the inferior alveolar canal. Adult skulls (n = 25) were radiographed using two panoramic machines both with and without radiopaque markers placed into the inferior alveolar canal and anterior loop. The anterior loop was also measured directly using flexible tubing (2 mm in diameter). Significantly fewer loops were detected in radiographs as compared with anatomic assessment (P < .001). A significant loop (> 2 mm) was identified in 92% to 96% of the direct measurements, whereas radiographs identified only 56% (Panelipse) and 76% (Orthoralix). Analysis of variance demonstrated that the skeletal length of the anterior loop was significantly greater than that measured radiographically (P < .0001). The average length of the anterior loop based on direct measurements was 6.95 mm, whereas radiographic measurements were 3.18 mm (Panelipse) and 3.45 mm (Orthoralix). The difference in detection of anterior loops among the radiographic procedures studied was not significant (P > .05). PMID- 8225465 TI - Occlusal force and mandibular movements in patients with removable overdentures and fixed prostheses supported by implants in the maxilla. AB - Thirteen patients with complete dentures in the maxilla were treated with implants to support a removable bar-retained overdenture for 1 year. Thereafter, the patients were provided with fixed prostheses and were followed up for another year. Maximal occlusal force and mandibular movements were recorded with and without the bar supporting the denture and then during the subsequent year with fixed prostheses. Significant changes in occlusal force capacity and mandibular movements were observed when the bar was removed. Less obvious functional changes were found when the fixed prostheses were placed, with the exception of occlusal force levels, which increased significantly over time. PMID- 8225466 TI - Light and confocal laser scanning microscopic evaluation of hydroxyapatite resorption patterns in medullary and cortical bone. AB - An animal study was conducted with unloaded blocks and hydroxyapatite (HA)-coated titanium implants. Four HA blocks were positioned in rabbit tibiae and four HA coated titanium implants were positioned in pig tibiae. Implants were positioned so that half was placed in cortical bone and half in medullary space. Biopsy specimens were taken 4 months after implant placement for histologic evaluation. Light microscopy and confocal laser scanning microscopy demonstrated that the HA resorption rate was higher in the medullary spaces, whereas resorption was almost absent in the areas embedded in cortical bone. PMID- 8225467 TI - Application of hydroxyapatite-coated implants as support for palatal lift prostheses in edentulous patients with cleft palate: a clinical report. AB - A palatal lift prosthesis (PLP) is a prosthetic speech aid used to compensate for palatopharyngeal incompetence. However, it requires dental abutments to facilitate its function. In two edentulous patients with cleft palate, hydroxyapatite-coated dental implants were used to increase PLP retention. In both cases, palatopharyngeal closure was obtained and PLP function was deemed effective, as determined by speech evaluation and nasal air flow measurements, for more than 3 years. This treatment is useful for enhancing the retention of a PLP in patients who have few or no remaining natural tooth abutments. PMID- 8225468 TI - A proposed radiographic index for assessment of the current status of osseointegration. AB - The longevity of an osseointegrated implant is directly affected by the maintenance of its osseous anchorage. Progressive marginal bone loss would be detrimental to its survival. This article proposes a new radiographic index for use in the radiographic assessment of peri-implant marginal bone levels. The index scores range from 0 to 6 and denote marginal bone loss in percentages relative to the implant body length measured from its coronal-most margin in an apical direction. The proposed index should be of significance in: (1) use of short implants (7 to 10 mm); (2) evaluation of implants functioning in areas of low bone density (types 3 and 4); (3) prognosis and treatment of ailing/failing implants; (4) follow-up of implants placed in extraction sites; and (5) correlation of crown-to-implant ratio with marginal bone loss. PMID- 8225469 TI - Hemorrhage of the floor of the mouth resulting from lingual perforation during implant placement: a clinical report. AB - The edentulous mandible provides a frequent indication for oral implants. In many cases implants are placed in the interforaminal region to support fixed prostheses or give retention to removable overdentures. One of the hazards of this seemingly safe surgical area is hemorrhage of the floor of the mouth. In this article, two cases of this life-threatening complication are reported. PMID- 8225470 TI - Ridge augmentation with titanium screws and guided tissue regeneration: technique and report of a case. AB - A technique is described for augmenting an edentulous ridge buccally with a mixture of freeze-dried bone and resorbable tricalcium phosphate covered with a Gore-Tex membrane. Titanium screws are used to prevent the collapse of the regenerative materials. In the patient described, the buccolingual dimensions of the edentulous mandibular ridge more than doubled, thus allowing the placement of two implant cylinders. The rationale for such a procedure is discussed. PMID- 8225471 TI - Should certified prosthodontic specialists be surgically placing dental implants? PMID- 8225472 TI - Detection of oxidative damage by 32P-postlabelling: 8-hydroxydeoxyguanosine as a marker of exposure. AB - Human exposure to reactive oxygen species is unavoidable and has been implicated in the etiology of a number of human diseases. This exposure results in the formation of various modified DNA bases: the promutagenic lesion 8 hydroxydeoxyguanosine (8OHdG), in particular, is a major product. We have developed an assay using ion-pair HPLC and 32P-postlabelling to quantify 8OHdG in human DNA with high specificity and sensitivity. An internal standard is used to account for variations in labelling efficiency. Chemically synthesized 8OHdG 3' monophosphate and 5'-monophosphate standards were used to optimize the HPLC-32P postlabelling and TLC separative steps, respectively. The assay was validated using known ratios of 8OHdG to normal nucleotides. The limit of detection is in the range of one 8OHdG residue per 10(6)-10(7) dG residues. Using this procedure, 8OHdG levels of 16-35 8OHdG adducts per 10(5) dG residues have been found in leukocytes isolated from patients who received 180-200 cGy of ionizing radiation. These levels were 2-4-fold greater than those found in an unexposed individual. Since 8OHdG may be formed during DNA extraction and digestion, current procedures for measuring background levels are discussed. PMID- 8225473 TI - 32P-postlabelling analysis of bulky aromatic adducts. AB - The 32P-postlabelling assay has emerged as a major tool for detecting DNA adducts induced by structurally diverse carcinogens, particularly bulky aromatics. The assay comprises enzymatic degradation of DNA to 3'-mononucleotides, enrichment of adducts, 5'-32P-labelling, adduct separation by TLC, and detection and quantitation of adducts. This report describes (1) an improved solvent extraction procedure for isolating DNA from tissues and (2) an up-dated version of the assay that we consider optimal for bulky adducts. The use of a non-urea solvent mixture (isopropanol: 4 M ammonium hydroxide) has been found to improve adduct separation and signal-to-noise ratios issues and (2) an up-dated version of the assay that we consider optimal for bulky adducts. The use of a non-urea solvent mixture (isopropanol: 4 M ammonium hydroxide) has been found to improve adduct separation and signal-to-noise ratios. PMID- 8225474 TI - Singlet oxygen-induced DNA damage: product analysis, studies of biological consequences and characterization of mutations. AB - The DNA lesions induced by free 1O2 and the biological and mutagenic consequences of 1O2-induced DNA damage have been studied. Using anion exchange HPLC, reverse phase HPLC with electrochemical detection and 32P-postlabelling methods, we have shown that 1O2 reacts with 2'-deoxyguanine 3'-monophosphate (dGp) but not with any other dNp. Reaction with dGp yields a large number of products; one minor product was identified as 7-hydro-8-oxo-2'-deoxyguanosine 3'-monophosphate (8-oxo dGp), and a second tentatively as a formamidopyridine derivative of dGp. 8-Oxo dGp was also found after reaction of 1O2 with single-stranded (ss) DNA, double stranded (ds) DNA or an oligonucleotide (16-mer) having one G. With the oligonucleotide we found a second unidentified reaction product. With ss DNA, 8 oxo-dG was a much more prominent product than in the reaction of 1O2 with free dGp and the yield was about eight-fold higher than with ds DNA. This agrees with our finding that ss M13 DNA is at least 100-fold more sensitive than ds M13 DNA to biological inactivation by 1O2. The inactivation of ss M13 DNA must be largely due to 1O2-induced lesions other than 8-oxo-dG. In agreement with the observed preferential reaction of 1O2 with dG, most of the mutations induced by 1O2 in ss or ds M13mp10 DNA occurred at a G or G/C basepair, respectively. A preference for G(C) to T(A) transversions was observed for which 8-oxo-dG might have been responsible. In ss DNA, a significant number of mutations are characterized by the fact that a G is deleted. PMID- 8225475 TI - 32P-postlabelling analysis of alkylphosphotriesters in DNA. AB - We have developed a highly sensitive 32P-postlabelling assay for the detection of alkylphosphotriesters in DNA using selective enzymatic DNA hydrolysis, sample preparation by C18 Sep-Pak solid phase extraction and product analysis by HPLC. The assay provides information on the quantity of the alkylphosphotriesters and gives some clues with respect to their identity. PMID- 8225476 TI - Characterization by 32P-postlabelling of 8-methoxypsoralen adducts. AB - 8-Methoxypsoralen is a bifunctional furocoumarin used in human photochemotherapy. It can form two kinds of DNA adduct, monoadducts and interstrand cross-links. These adducts have been detected by 32P-postlabelling using hydrolysis with DNase1, alkaline phosphatase and snake venom phosphodiesterase, and longer labelling than usual, with more T4 polynucleotide kinase. Using preliminary two dimensional chromatography (D1, D2) followed by transfer of adducts to separate PEI cellulose sheets for further development (D3, D4), we observed three spots corresponding to the adducts sought. Two experiments (dose-effects and shift of radioactivity) have confirmed the origin of the three spots. PMID- 8225477 TI - Ochratoxin A-related DNA adducts in urinary tract tumours of Bulgarian subjects. AB - Ochratoxin A (OTA), a natural contaminant of mouldy food and feed, is suspected of being one of the etiological agents responsible for Balkan endemic nephropathy (BEN) and the associated urinary tract tumours. We have previously shown that ochratoxin A is genotoxic as expressed by DNA single-strand breaks. DNA-OTA adducts have been detected in various mouse organs after ochratoxin A treatment. Tumorous tissues from three kidneys and five bladders of Bulgarian patients undergoing surgery for cancer and from three non-malignant kidneys collected from French subjects were analysed for DNA adducts. Several adducts with the same RF values as those obtained from mouse kidney after treatment with OTA (one major and some minor adducts) were detected, mainly in kidney but also in bladder tissues from Bulgaria. No adducts were detected in French kidney tissues. These results provide new evidence of the possible role of OTA in the development of tumours of the urinary tract in Bulgaria. PMID- 8225478 TI - 32P-postlabelling in studies of hormonal carcinogenesis. AB - The nature of the covalent modification of DNA by estrogens has been investigated by 32P-postlabelling and other techniques. Three different classes of covalent DNA alteration are induced by estrogens in vivo and are evaluated in this review for their potential to play a role in hormonal carcinogenesis: (a) estrogen-DNA adducts formed by reactive estrogen quinone metabolites; (b) enhancement of endogenous DNA modification by estrogen specifically in the target organ for cancer; (c) 8-hydroxyguanine bases formed by free radicals generated by redox cycling of estrogen. Additional studies are required to identify the class or classes of DNA adducts that are crucial for the induction of hormonal cancer. PMID- 8225479 TI - I-compounds--endogenous DNA markers of nutritional status, ageing, tumour promotion and carcinogenesis. AB - Certain bulky DNA modifications increase with age in laboratory animals without known exposure to carcinogens and are known as indigenous or I-compounds. Most of the spots detected on TLC maps by the nuclease P1-enhanced bisphosphate version or the monophosphate version of the 32P-postlabelling assay fit this definition of I-compounds. Chromatographic profiles and levels of these nucleotides greatly depend on animal species, strain, tissue and gender, but are also influenced by the diet and by chemical exposures. Thus, both genetic and environmental factors contribute to the formation of I-compounds. It appears that they are derived from endogenous DNA-reactive compounds generated during normal nutrient metabolism and that regulatory mechanisms exist which control their levels in tissue DNA. PMID- 8225480 TI - Formation and persistence of specific purine DNA adducts by 32P-postlabelling in target and non-target organs of rats treated with aristolochic acid I. AB - Binding of the naturally occurring carcinogen, aristolochic acid I (AAI), to DNA in male Wistar rats has been examined. Rats were treated orally with a single dose (13.8 mmol) of AAI and sacrificed 1 day and 1, 2, 4, 16 and 36 weeks after treatment. The formation and persistence of two specific purine AAI-DNA adducts were studied utilizing the nuclease P1 enhancement method of the 32P postlabelling assay. Both adducts, 7-(deoxyadenosin-N6-yl)-aristolactam I (dA AAI) and 7-(deoxyguanosin-N2-yl)-aristolactam I (dG-AAI), were detectable for up to 36 weeks in all target (forestomach) and non-target (glandular stomach, liver, lung, urinary bladder) organs and showed differential rates of removal and persistence. In all organs, dA-AAI was the major adduct present. In the target organ (forestomach), both adducts were removed rapidly within the first two weeks; thereafter, extensive removal of dG-AAI continued, whereas dA-AAI attained constant levels (4-36 weeks). Of interest was the marked decrease of both adducts in glandular stomach, the neighbouring non-target tissue to the forestomach. These results suggest that the persistence of a specific AAI-DNA adduct, namely dA-AAI, in the target organ may be the critical lesion responsible for initiation of carcinogenesis by AAI. PMID- 8225481 TI - 32P-postlabelling studies on the DNA adducts of the food mutagens/carcinogens IQ and PhIP--adduct formation in a chemical system, and by rat and human metabolism. AB - The DNA adducts of the food mutagens/carcinogens IQ and PhIP were studied by means of 32P-postlabelling techniques. Adducts were generated in vitro and in vivo by three techniques: by photolysis of azido-IQ and azido-PhIP in the presence of dGp, calf-thymus DNA or Salmonella; by administration of IQ and PhIP to rats; and by incubation of cultured COS-1 cells with IQ. These cells expressed the human cytochromes P450 1A1 or P450 1A2 and/or the human N-acetyltransferases NAT1 or NAT2. The data demonstrate that, in the photolytic, rat and human systems, a common IQ metabolite and a common PhIP metabolite are formed together with common sets of IQ adducts and PhIP adducts. The data obtained in the human system show that N-hydroxy-IQ, formed by cytochrome P450, binds poorly to DNA, whereas more efficient binding occurs in the presence of NAT1 and most efficient binding in presence of NAT2. This indicates an O-acetyltransferase activity of human NAT1 and NAT2 and formation of N-acetoxy-IQ as an intermediate and immediate precursor of the ultimate arylnitrenium ion. The effect of the polymorphic NAT2 suggests a critical role for the human acetylation polymorphism in the DNA-binding of IQ in humans and in its genotoxic implications. PMID- 8225482 TI - Application of the 32P-postlabelling assay to the inhibition of 2-amino-3 methylimidazo[4,5-f]quinoline (IQ)-DNA adduct formation by dietary fatty acids. AB - The potent heterocyclic food mutagen IQ is carcinogenic in the CDF1 mouse, affecting the liver, lung and forestomach. Using 32P-postlabelling methods we have isolated up to five IQ-DNA adducts from both target and non-target organs after oral administration of IQ. Up to four additional non-specific adducts, found when the 32P-postlabelling assay was run under intensification conditions, could be attributed to the use of a certain brand of microcentrifuge tube in the assay. CLA is a mixture of heat-generated derivatives of linoleic acid, each with a conjugated double bond system, that has chemopreventive properties in rodents. To examine the effect of CLA on the formation of IQ-DNA adducts in CDF1 mice, we administered CLA by gavage every other day for 45 days, followed by a single oral dose (50 mg/kg) of IQ. Tissues collected 24 h later were analysed for IQ-DNA adducts by 32P-postlabelling. Compared to controls, CLA treatment caused inhibition of adduct formation in the liver, lung, large intestine and kidney. In the kidneys of females, CLA treatment inhibited IQ-DNA adduct formation almost completely (95.2%). Male F344 rats were fed a control diet or an isocaloric diet containing 20% menhaden oil (MO), a rich source of omega-3 fatty acids, for six weeks, then given a single oral dose of IQ. Analysis for IQ-DNA adducts by 32P postlabelling one or six days later revealed that on day 1 the MO diet caused a 6 7 fold decrease in adduct formation in the liver and an up to 2-fold decrease in both the small and large intestine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225483 TI - Formation of DNA adducts from 1-hydroxymethylpyrene in liver cells in vivo and in vitro. AB - The binding of 1-hydroxymethylpyrene (HMP) and its active metabolites, 1 hydroxymethylpyrene sulfate (SMP) and 1-chloromethylpyrene (CMP), to DNA was studied. In the liver of rats, maximum adduct levels were observed 1.5 h after i.p. injection of HMP, followed by a relatively rapid decrease. Separate exposure of different liver cell types in vitro to HMP led to high adduct levels in parenchymal cells, intermediate levels (1/10) in endothelial cells and low levels (1/200) in Kupffer cells. The adduct patterns were similar in the different cell types. The same pattern was also obtained when isolated DNA was incubated with SMP or CMP. One of the four major spots co-chromatographed on TLC with a dG adduct, one with a dA adduct and one with both dG and dA adducts. The fourth spot did not co-chromatograph with any of the adducts observed in reactions with any nucleic acid homopolymer. PMID- 8225484 TI - Biomonitoring of human exposure to carcinogenic nitroaromatic compounds: a pilot study on DNA adduct formation by 1,6-dinitropyrene in rats. AB - DNA adduct formation by 1,6-dinitropyrene (DNP) was examined in various rat tissues. Intraperitoneal administration of 0.2, 1.0 or 5.0 mg DNP/kg b.w. caused the formation of one major and, at the higher doses, two minor DNA adducts. The highest level of about 300 adducts/10(9) nucleotides was found in urinary bladder DNA. 5-10-fold lower adduct levels were found in the DNA of white blood cells (WBC), liver, lung and small intestinal mucosa. DNA adduct levels in the bladder were highest in Sprague-Dawley males followed by Sprague-Dawley females and Wistar males. Administration by gavage was less effective than intraperitoneal injection. Intratracheal instillation of microcrystalline DNA suspensions did not lead to any detectable adduct formation. The results indicate that WBC DNA may not be a reliable DNA source for biomonitoring human exposure to nitroarenes and that nitroarene-DNA adducts may only be detectable at high levels of exposure. PMID- 8225485 TI - An improved 32P-postlabelling assay for detection and quantitation of styrene 7,8 oxide-DNA adducts. AB - Using DNA modified with [7-3H]styrene 7,8-oxide (SO) in vitro we have standardized the 32P-postlabelling assay for detecting SO-DNA adducts. Nuclease P1-enriched adducts were 32P-labelled and purified by high-salt (4.0 M ammonium formate, pH 6.1) C18 reverse-phase TLC. After elution from the layer with 2 butoxyethanol:H2O (4:6), adducts were separated by two-dimensional PEI cellulose TLC in non-urea solvents (2.0 M ammonium formate, pH 3.5, and 2.7 M sodium phosphate, pH 5.6). One major, three minor and several trace adducts were detected. The efficiency of the kinase reaction depended on the ATP concentration. Use of standard labelling conditions ([gamma-32P]ATP, < or = 3000 Ci/mmol; < or = 2 microM) resulted in poor (4-7%) adduct recovery. An ATP concentration of 40 microM, however, increased the labelling efficiency by a factor of 5-8 (35-55%) based on 3H-SO labelled DNA). The results indicate that the new separation technique is suitable for the relatively polar SO-DNA adducts and that high labelling efficiency can be achieved. PMID- 8225486 TI - 32P-postlabelling analysis of DNA-adducts formed by 4-(N-carbethoxy-N nitrosamino)-(3-pyridyl)-1-butanone and 4-oxo-4-(3-pyridyl)- butanal. AB - The tobacco-specific nitrosamine, 4-(N-nitrosomethylamino)-1-(3-pyridyl)- 1 butanone (NNK) can be activated to DNA-binding species by two pathways: methylene hydroxylation leads to an equimolar formation of methyldiazohydroxide and 4-oxo-4 (3-pyridyl)-butanal followed by DNA methylation, while methyl hydroxylation leads to DNA pyridyloxobutylation via the 4-(3-pyridyl)-4-oxobutyldiazohydroxide (Hecht et al., 1988; Spratt et al., 1989). We report here that the ketoaldehyde, 4-oxo-4 (3-pyridyl)-butanal, as well as 4-(N-carbethoxy-N-nitrosamino)-1-(3- pyridyl)-1 butanone (NNC), a model compound for pyridyloxobutylation, form DNA adducts in vitro, as detected by 32P-postlabelling assays. The two major adduct spots of both compounds were chromatographically indistinguishable, suggesting structurally similar DNA adducts. PMID- 8225487 TI - Effect of tissue fixation on recovery of DNA adducts in the 32P-postlabelling assay. AB - The suitability of fixed tissue as a source of DNA for 32P-postlabelling studies on DNA adduct formation has been investigated. Tissues (spleen, liver, lung, colon and kidney) from rats treated i.p. with benzo[a]pyrene (BaP) (100 mg/kg) or 2-acetylaminofluorene (AAF) (40 mg/kg) were removed and placed in formalin fixative for 1, 7, 28 or 92 days. DNA was isolated and 32P-postlabelled with nuclease P1 enhancement for BaP-modified DNA and with butanol enhancement for AAF modified DNA. There was a marked loss of adducts with time of fixation, most noticeably between 1 and 7 days. DNA from tissue that had been fixed for 92 days and then wax-embedded had similar levels of adducts to the 92-day fixed-only samples. Further tissue samples were fixed for 24 h with formalin, modified Methacarn or Bouin's and then wax-embedded. DNA was then extracted either immediately or 28 days later. Adducts were recovered from formalin- and Methacarn treated tissues at levels comparable to those detected in DNA from frozen tissues and levels were stable once the tissues were wax-embedded. However, DNA recovery from Bouin's-fixed tissue was poor. The results indicate that only tissues that have been wax-embedded after the minimum required fixation period are suitable sources of DNA for 32P-postlabelling analysis. PMID- 8225488 TI - Analysis of polycyclic aromatic hydrocarbon-DNA adducts by postlabelling with the weak beta-emitters 35S-phosphorothioate and 33P-phosphate, immobilized boronate chromatography and high-performance liquid chromatography. AB - The wide range of carcinogenic activities of optical isomers of bay-region diol epoxides of polycyclic aromatic hydrocarbons (PAHs) demands analytical techniques capable of distinguishing both the diastereoisomer and enantiomers of the PAH diol-epoxide (PAH-DE) responsible for DNA adduct formation. Our laboratory recently developed postlabelling procedures using the radioisotope 35S for HPLC analysis of benzo[a]pyrene (BaP)-DNA adducts formed in BaP-treated hamster cell cultures. To allow identification and complete separation of anti-PAH-DE-DNA adducts from syn-PAH-DE-DNA adducts, an immobilized boronate chromatography procedure was developed using a high-capacity boronate column. Analysis of 7,12 dimethyl-benz[a]anthracene (DMBA)-DNA adducts formed in treated hamster embryo cells by this boronate chromatography procedure and 35S-postlabelling demonstrated that both anti- and syn-DMBA-DE were bound to DNA. These analytical techniques also provided evidence that the potent carcinogen and environmental PAH dibenzo[a,l]pyrene was metabolized in cells to a DNA-binding intermediate with structural characteristics of an anti-PAH-DE. Conditions were also developed for using 33P-labelled ATP in the postlabelling assay. The use of [33P] instead of [35S]ATP offers the advantages of ease of labelling and greater sensitivity while still using a weak beta-emitter. The development of PAH-DNA adduct analysis techniques with improved sensitivity allows more detailed studies of how PAHs interact with DNA and how this leads to cancer induction. PMID- 8225489 TI - Fluorescent postlabelling of modified DNA bases. AB - Postlabelling of modified DNA bases by methods other than the 32P method has attracted little attention to date. Three approaches using fluorescence have been made: (a) detection of a characteristic adduct-DNA functionality, e.g. reaction of cis-vicinal diols of PAH-DNA adducts with a boronate-containing fluorescent molecule; (b) reaction of modified nucleotides with a reactive fluorescent molecule, e.g. detection of 8-HOdG-5'-monophosphate by reaction with dansyl chloride; (c) reaction of modified DNA bases with reagents giving fluorescent products, e.g. derivatization of 7-MeGua with phenylmalondialdehyde (PhMal). Recent results in developing the latter approach are described. Reaction of a series of 7-alkylguanines, including 7-methyl-, 7-ethyl-, 7-(2-hydroxyethyl)-, 7 carboxymethyl-, 7-(2-carboxyethyl)- and 7-(2,3-dihydroxypropyl)-guanine, with PhMal gave fluorescent products showing similar spectral properties. The detection limit for fluorescence detection following reverse-phase HPLC separation was typically less than 1 pmol injected. In order to improve the sample clean-up before analysis, an antiserum has been prepared against the fluorescent derivative of 7-(2-carboxyethyl)guanine bound to carrier protein via the carboxyl group. The resulting antiserum cross-reacted with a number of PhMal derivatives. PMID- 8225490 TI - Separation of 32P-labelled nucleoside 3',5'-bisphosphate adducts by HPLC. AB - Relatively few reported attempts have been made to substitute HPLC for the thin layer ion-exchange chromatography (TLC) conventionally used in the 32P postlabelling assay. Using a reverse-phase phenyl-modified silica gel column and a gradient of methanol in 0.5 M sodium phosphate buffer (pH 2.0), we were able to improve the resolution of very similar adducts. Combined with on-line detection of Cerenkov radiation, this method allows separation of sub-femtomole quantities of 32P-labelled nucleoside 3',5'-bisphosphates modified by bulky carcinogens. Using this method, we were able to separate nine of the ten major adducts formed by reaction of the diol-epoxides of ten polycyclic aromatic hydrocarbons with DNA, and resolve different adducts formed by a single carcinogen. The major adducts formed by benzo[b]fluoranthene (BbF) or dibenz[a,h]anthracene in mouse skin in vivo have been shown to be distinct from the adducts formed directly by the bay-region diol-epoxides. The heterocyclic amines IQ and MeIQ have each been shown to form one major DNA adduct in several in vitro and in vivo systems; using HPLC we were able to resolve the two adducts formed by these food mutagens. HPLC is especially useful for the identification of adducts by means of chromatographic comparisons and in the analysis of the multiple adducts formed by complex mixtures of environmental carcinogens. The major adducts formed by benzo[a]pyrene (BaP) and BbF in mouse skin in vivo that were not resolved on TLC were well separated by HPLC and thus a major DNA adduct formed in the skin of mice treated topically with coal tar was found to be derived from BaP rather than BbF. PMID- 8225491 TI - Combined micropreparative techniques with synchronous fluorescence spectroscopy or 32P-postlabelling assay for carcinogen-DNA adduct determination. AB - Methods for the detection of carcinogen-DNA adducts require both sensitivity and specificity. The 32P-postlabelling assay is highly sensitive but lacks adduct specificity. The strategy reported herein combines micropreparative techniques including HPLC and immunoaffinity chromatography (IAC) to enhance chemical specificity. The resultant assays have retained sensitivity for human DNA analysis. Combined HPLC and 32P-postlabelling has detected 7-methyldeoxyguanosine in human lung samples, while combined IAC and 32P-postlabelling has detected O6 methyldeoxyguanosine adducts in stomach tissue. The limits of detection are one adduct in 10(7) and 10(8) unmodified deoxyguanosine (dG), respectively. IAC was combined with 32P-postlabelling to detect polycyclic aromatic hydrocarbon (PAH) DNA adducts in human lung. The detection limit was one adduct in 10(7) dG. Our laboratory has also employed synchronous fluorescence spectroscopy (SFS) for the detection of adducts formed from benzo[a]pyrene in human lung. Complex fluorescence matrices suggest the presence of other PAH-DNA adducts. Both the SFS assay and the 32P-postlabelling assay were applied to a series of human samples and a high correlation was found for adduct levels. The development of such assays using synthetic standards, internal standards, determination of calibration curves and validation with corroborating methods is required for chemically specific and sensitive adduct detection. PMID- 8225492 TI - 32P-postlabelling analysis of small aromatic and of bulky non-aromatic DNA adducts. AB - The 32P-postlabelling methodology for analysis of DNA adducts derived from carcinogens containing one aromatic ring (e.g., safrole, styrene oxide, benzene metabolites, 1-nitrosoindole-3-acetonitrile) or a bulky non-aromatic moiety (e.g., mitomycin C, diaziquone) is reviewed. Six steps are involved: digestion of DNA to 3'-nucleotides, enrichment of adducts, 32P-labelling of adducts, separation of labelled adducts by TLC, detection, and quantitation. The first step, DNA digestion with micrococcal nuclease and spleen phosphodiesterase, is applicable to DNA modified with most carcinogens independent of their size and structure. Of the two commonly used procedures for enrichment of aromatic adducts in DNA digests, the nuclease P1 treatment is substantially more effective than butanol extraction for small aromatic and bulky non-aromatic adducts. For initial purification of these adducts from unadducted material after 32P-labelling, multi directional polyethyleneimine (PEI)-cellulose TLC using 1 M sodium phosphate, pH 6.0, as the D1 solvent is not suitable, because they are not retained on PEI cellulose under these conditions. A higher concentration of sodium phosphate (e.g., 2.3 M) or development with D1 and D3 solvents in the same direction helps to retain adducts of safrole and of benzene metabolites. Also, transfer of adducts from multiple cut-outs above the origin after D1 chromatography, as adopted for analysis of I-compounds, is potentially applicable. However, initial purification by reverse-phase TLC, followed by in situ transfer and resolution by PEI-cellulose TLC has been found to be most effective for these adducts. Reverse phase TLC at 4 degrees C or in a stronger salt solution further improves retention of some adducts (e.g., mitomycin C and diaziquone adducts). For adduct separation by PEI-cellulose TLC, salt solutions with or without urea are used. PMID- 8225493 TI - 32P-postlabelling methods for cyclic DNA adducts. AB - 32P-Postlabelling procedures coupled with HPLC have been developed to detect and measure a range of cyclic DNA adducts formed by bifunctional genotoxic agents. The methods are based on reverse-phase HPLC, particularly column-switching HPLC, to enrich adduct 3'-monophosphates before labelling. Following 3' dephosphorylation of the 3'5'-[5'-32P]bisphosphates with nuclease P1, the resulting 5'-[32P]monophosphate adducts are resolved, identified and characterized by co-chromatography with synthetic reference standards. The procedures have been applied to a number of cyclic adducts including those formed by chloroacetaldehyde, glycidaldehyde and malonaldehyde. In general, labelling efficiencies measured as chromatographed 5'-[32P]monophosphates were in the range 30-40%. However, the values for the malonaldehyde deoxyguanosine adduct were much lower. The techniques have been applied to studies on the formation of DNA adducts in the skin of male C3H mice treated cutaneously with glycidaldehyde. The HPLC-32P-postlabelling analysis of epidermal DNA hydrolysates indicated that a single major cyclic adduct was formed by reaction with deoxyadenosine residues in mouse skin DNA. The adduct was identified as a hydroxymethyl ethenodeoxyadenosine adduct by comparison with a synthetic standard. This adduct was highly fluorescent and it was possible to make quantitative comparisons of the amounts of adduct determined by either HPLC-32P-postlabelling or HPLC-fluorescence detection. PMID- 8225494 TI - Immunoaffinity clean-up combined with 32P-postlabelling analysis of 1,N6 ethenoadenine and 3,N4-ethenocytosine in DNA. AB - Immunoaffinity gels able to bind 1,N6-ethenodeoxyadenosine 3'-monophosphate (3' epsilon dAMP) and 3,N4-ethenodeoxycytidine 3'-monophosphate (3'-epsilon dCMP) were prepared. These gels retained their specific binding capacity for 3'-epsilon dAMP or 3'-epsilon dCMP in the presence of a large excess (> 2 mg per column) of unmodified 3'-deoxynucleotide monophosphates. 3'-epsilon dAMP and 3'-epsilon dCMP were 32P-postlabelled in the presence of [gamma-32P]ATP and T4 polynucleotide kinase to give either 3',5'-[5'-32P]-bisphosphates or 5'-[32P]monophosphates. 3',5'-[5'-32P]Bisphosphates were recovered from the labelling mixture by immunoprecipitation and quantitated by Cerenkov counting (method A). The detection limit of this procedure was 1 fmol with an efficiency of 80% for both ethenonucleotides. Alternatively, 5'-[32P]epsilon dAMP and 5'-[32P]epsilon dCMP were analysed by two-dimensional TLC on PEI cellulose and autoradiography (method B). The detection limit of method B was 50 amol of ethenonucleotide. Methods A and B are complementary and can quantify the formation and repair of 3'-epsilon dAMP and 3'-epsilon dCMP in DNA from rats exposed to a low level of vinyl chloride. PMID- 8225495 TI - Measurement of oxidative base damage to DNA by using HPLC-32P-postlabelling and GC/MS-selective ion monitoring assays. AB - A 32P-postlabelling assay has been developed for singling out specific oxidized base lesions. Emphasis was placed on the quantitative aspect and the accuracy of the assay, which require the use of calibration curves and microreactions, respectively. The method was successfully applied to the detection and the measurement of adenine N1-oxide and 5-hydroxymethyluracil in cells exposed to agents inducing oxidative stress including H2O2 and UV-A radiation. The sensitivity of the assay allows the detection of one lesion in 10(6) normal bases in 1 microgram of DNA. The GC/MS method when coupled to the selective ion monitoring (SIM) technique is about twenty times less sensitive, even for suitable substrates such as 5-hydroxymethyluracil and 5-hydroxyuracil, than the 32P-postlabelling assay. However, the former assay is much easier to apply, even though a derivatization step is necessary, and provides unambiguous structural information on the compound to be measured. Accurate quantitative measurements can be obtained when stable, isotopically labelled standards are available. PMID- 8225496 TI - Detection by HPLC-32P-postlabelling of DNA adducts formed after exposure to epoxybutene and diepoxybutane. AB - We have developed an HPLC-32P-postlabelling procedure to detect DNA adducts formed by epoxybutene and diepoxybutane. The method exploits the interaction of the two epoxides with deoxynucleotides and polydeoxynucleotides to optimize the HPLC enrichment of adducted nucleotides before 32P-postlabelling. Using this approach, a number of guanine adducts were identified after the exposure of dGMP, poly(dG-dC) or calf thymus DNA to epoxybutene and diepoxybutane, and a major adenine adduct was identified in poly(dA-dT) and calf thymus DNA exposed to diepoxybutane. PMID- 8225497 TI - DNA adduct profiles and levels in placenta, blood and lung in relation to cigarette smoking and smoky coal emissions. AB - Tobacco smoking and indoor smoky coal combustion emissions lead to exposures to complex environmental mixtures that have been associated with increased lung cancer mortality rates in the USA and Xuan Wei County, China, respectively. Human exposures to benzo[a[pyrene (BaP) present in smoky coal emissions are 20-200 times greater than exposure to BaP from smoking 1-2 packs of cigarettes per day. We compared DNA adducts resulting from cigarette smoking and coal smoke exposures in blood cells, placental syncytial nuclei, whole placental tissue homogenates and lung cells. Postlabelling analysis of DNA isolated from placental tissue of smokers revealed multiple adducts as a diagonal radioactive zone. DNA adduct levels for both white blood cells (WBC) and lymphocyte DNA were approximately 2.5 fold higher in smokers than in non-smokers. One smoking-related adduct was detected in both WBC and lymphocyte DNA, but no exposure-related adducts were detected in either WBC DNA or placental tissue DNA isolated from coal smoke exposed individuals, despite higher exposure to BaP and aromatic organics. In contrast, discrete adducts unrelated to smoking status were detected, by 32P postlabelling analysis, in DNA from syncytial nuclei isolated from placental villus samples from smokers and non-smokers. One major placental syncytial adduct co-migrated with a major BaP-derived DNA adduct. DNA adduct levels were approximately four-fold higher in bronchoalveolar lavage (BAL) cells from coal smoke-exposed individuals compared to unexposed individuals and suggest that DNA reactive intermediates concentrate in the respiratory tract and may not be detectable in whole WBC or placental tissue homogenates. Our findings also suggest that the distribution and concentration of placental DNA adducts may be dependent on the site at which the placenta is sampled. PMID- 8225498 TI - Studies on DNA adduction with heterocyclic amines by accelerator mass spectrometry: a new technique for tracing isotope-labelled DNA adduction. AB - DNA adduction in rodents at doses equivalent to human dietary exposure (10(4) 10(6)-fold lower than laboratory studies) is being studied using accelerator mass spectrometry (AMS). AMS is a nuclear physics technique for detection of cosmogenic isotopes and has been used for specifically selecting and counting 14C. Using AMS, DNA adducts are detectable at levels of 1-10 adducts/10(12) nucleotides following acute and chronic dosing regimes with 14C-labelled carcinogens. The adduct detection limit has been imposed by the natural abundance of 14C in the samples and animal-to-animal variation. AMS is also being coupled to HPLC, multidimensional TLC and radio-immunoassay. In addition, AMS's great sensitivity makes it useful for demonstrating that drugs and chemicals do not bind to DNA. The use of AMS, however, is limited to situations where radiolabelled agents can be used. The data suggest that AMS is extremely useful in obtaining quantitative data on the effects of carcinogens on DNA at the low doses common for actual human exposures and may be useful in human studies. PMID- 8225499 TI - Monitoring tobacco smoke-induced DNA damage by 32P-postlabelling. AB - Adduct levels in smokers are substantial, i.e. about 1 adduct in 10(7) nucleotides in lung DNA of lung cancer patients. Tobacco smoke-associated (TSA) DNA adducts, formed in a dose- and exposure time-dependent manner in smokers' lung, persist for considerable periods of time, consistent with epidemiological evidence showing quantitative associations of smoking with lung cancer risk and mortality. These results support a causal relationship between such adducts and tobacco smoking-induced cancer. TSA adducts were also present in various other target organs for tobacco-smoke carcinogenesis, including kidney, bladder, oesophagus and pancreas. PMID- 8225500 TI - Determination of smoking-related DNA adducts in lung-cancer and non-cancer patients. AB - Aromatic DNA adduct levels were determined in macroscopically normal bronchial tissues from 98 patients undergoing pulmonary surgery. The mean DNA adduct level for the 45 current smokers was significantly higher than that of the 16 life-time non-smokers. There was a weak association between adduct levels and daily cigarette consumption above 10 cigarettes per day. DNA adduct levels in the 37 former smokers suggested an exponential form of adduct elimination with a rapid initial and a slower later phase after cessation of smoking. There was no quantitative association between bronchial DNA adduct levels and lung cancer. PMID- 8225501 TI - Comparison of total DNA adduct levels induced in mouse tissues and human skin by mainstream and sidestream cigarette smoke condensates. AB - Cigarette smoke condensates (CSCs) of both mainstream (MS) and sidestream (SS) smoke were used to treat mice topically in equivalent amounts. Human skin maintained in short-term culture was also treated with the condensates. DNA adducts, induced by the CSCs and detected by the nuclease P1 method of 32P postlabelling, were quantified in a number of murine tissues and in the human skin DNA. In the five mouse tissues studied both MS-CSC and SS-CSC produced characteristic diagonal radioactive zones on TLC, indicative of the formation of multiple DNA adducts. In three tissues (skin, lung and kidney), SS-CSC induced greater total adduct levels than MS-CSC (statistically significant in skin and kidney, p < 0.05). However, greater adduct levels induced by MS-CSC were recorded for heart and bladder DNA (not statistically significant). Similar results to those found in mouse skin were obtained with human skin; SS-CSC induced a approximately 2-fold greater level of DNA adducts than MS-CSC (p < 0.05). Incubation of DNA directly with condensates in vitro demonstrated that DNA adducts could be formed without an exogenous metabolizing system. This direct interaction of condensates with DNA occurred at similar levels for both MS- and SS-CSC, although inclusion of an oxygen radical-generating system enhanced the SS CSC binding to a greater extent than that of the MS-CSC. PMID- 8225502 TI - Smoking-related DNA adducts in human cervical biopsies. AB - Cervical biopsy samples were taken from 35 women (19 smokers, 5 ex-smokers and 11 non-smokers) aged between 30 and 72, undergoing hysterectomy. DNA was isolated and analysed using 32P-postlabelling, with butanol extraction enhancement of the adducts. Resolution of the adducts was by thin-layer chromatography on polyethyleneimine (PEI)-cellulose. The pattern of adducts was similar to that of smoking-related adducts detected in other tissues and consisted mainly of a diagonal zone of radioactivity. The levels of adducts in DNA from the 19 smokers and a recent ex-smoker ranged from 1.93 to 6.04 adducts/10(8) nucleotides (mean = 3.67, SD = 1.36), in DNA from non-smokers from 1.35 to 3.98 (mean = 2.10, SD = 0.79) and in samples from ex-smokers from 2.57 to 3.35 (mean = 2.86), SD = 0.32). The increase in adduct levels in smokers compared with non-smokers was highly significant (Mann-Whitney test p = 0.002, one-tailed). These results are consistent with the hypothesis that smoking-related cervical cancer results from exposure to genotoxic components of cigarette smoke that become activated to DNA binding products in this tissue. PMID- 8225503 TI - Analysis of DNA adducts in smokers' lung and urothelium by 32P-postlabelling: metabolic phenotype dependence and comparisons with other exposure markers. AB - Carcinogen-DNA adduct levels in lung parenchyma (surgical specimens) and urothelial (exfoliated) cells of smokers, ex-smokers and non-smokers were investigated. DNA adducts were analysed by 32P-postlabelling and levels were compared with tissue-specific activity of cytochrome P450-related enzymes, or whenever possible, with metabolic phenotypes and other macromolecular adducts. Lung cancer patients who were recent smokers had significantly induced benzo[a]pyrene (BaP)-3-hydroxylase (AHH) and ethoxycoumarin O-deethylase activities in lung parenchyma compared with smoking non-cancer patients. Pulmonary AHH activity showed a good correlation with the intensity of immunohistochemical staining for P4501A(1). In lung cancer patients from Italy and Finland who were recent smokers, lung AHH activity was positively correlated (r approximately 0.65; p < 0.001) with bulky DNA adduct levels. In some lung DNA samples from smokers, the level of BaP-diol-epoxide adducts determined by HPLC with fluorescence detection showed significant positive correlation with lung AHH activity and bulky DNA adduct levels. Molecular dosimetry studies provided evidence that aromatic amines such as 4-aminobiphenyl (ABP) in tobacco smoke are primarily responsible for bladder cancer in smokers. The N-(deoxyguanosin-8-yl)-4 ABP adduct was the major smoking-related adduct in DNA of bladder biopsies from bladder cancer patients and in the DNA of exfoliated urothelial cells of smoking volunteers. The adduct levels of ABP with haemoglobin and with deoxyguanosine in urothelial DNA (determined by 32P-postlabelling) were linearly and significantly correlated, and both were related to recent cigarette smoking. Metabolic phenotype (fast/slow N-acetylator and N-oxidizer) significantly affected the levels of ABP-haemoglobin adducts. PMID- 8225504 TI - Monitoring occupational exposure to carcinogens. AB - 32P-Postlabelling has been used for biomonitoring occupational exposure to complex mixtures of polycyclic aromatic hydrocarbons in iron foundries, coke oven and aluminium plants and among roofers and surface-coating workers. Enhanced levels of aromatic DNA adducts have been detected in exposed populations in comparison to controls. Dose-related adduct formation has been found in iron foundry and coke-oven workers and roofers. The importance of longitudinal biomonitoring has been shown in two aluminium plants. Comparison between 32P postlabelling and immunoassays revealed wide variations. DNA adduct levels obtained by the current methods should thus be regarded as relative values between individuals and control and exposure groups. PMID- 8225505 TI - 32P-postlabelling analysis of human tissues and its potential use for biomonitoring. AB - Samples of clinically normal oral tissue were obtained from 17 tobacco smokers and 7 non- or ex-smokers undergoing surgery for intra-oral squamous cell carcinoma. Isolated DNA was analysed for the presence of aromatic DNA adducts using the 32P-postlabelling technique with adduct enhancement by either butanol extraction or nuclease P1 enrichment. DNA adduction detected following butanol extraction was more diverse and at a higher level than obtained with the P1 method. Adduct levels in the smokers and non- or ex-smokers were 1163 +/- 375 and 774 +/- 318 amol/micrograms, respectively. This difference is statistically significant (p < 0.05). The differential enhancement of adducts with the two protocols suggested that arylamines may be the source of at least a proportion of the DNA adduction detected. These data indicate that oral tissues are likely to be a suitable source of material for human biomonitoring and furthermore they highlight the importance of utilizing more than one enhancement procedure when examining DNA adduction induced by complex mixtures such as tobacco smoke or those encountered at industrial plants. PMID- 8225506 TI - Results of an interlaboratory trial of 32P-postlabelling. AB - An interlaboratory 32P-postlabelling trial involving 15 independent investigators was conducted. Each participant was provided with four DNA samples: 1, control DNA; 2, benzo[a]pyrene-modified DNA from mouse skin; 3, DNA from the lung of a tobacco smoker; 4, 2-acetylaminofluorene-modified DNA from mouse liver. The participants analysed the DNA samples by 32P-postlabelling using the standard procedure and using two procedures, nuclease P1 digestion and butanol extraction, to enhance the sensitivity of the assay. The qualitative and quantitative reproducibility of the results obtained by the different investigators are presented. A workshop discussion on postlabelling procedures is summarized. PMID- 8225507 TI - 32P-postlabelling studies of target tissues and bile from patients with familial adenomatous polyposis and from unaffected controls. AB - Patients with the inherited form of colon cancer, familial adenomatous polyposis (FAP), are at high risk of developing duodenal adenomas and carcinomas. The periampullary clustering of these neoplasms suggests that bile plays a role in their development. We investigated this theory using 32P-postlabelling to detect DNA adducts. We found significantly higher adduct levels in duodenum of FAP patients than in unaffected controls, and higher levels in duodenum than stomach. Levels of adducts were significantly higher in the small bowel of rats gavaged with FAP gallbladder bile than in the small bowel of those that received control gallbladder bile. We found that bile from FAP gallbladder produced significantly more DNA adducts than control gallbladder bile when incubated with salmon sperm DNA in vitro. These results support the hypothesis that the bile of FAP patients may be involved in the development of duodenal adenomas in these patients. PMID- 8225508 TI - Biomonitoring of aquatic systems. AB - The 32P-postlabelling analysis provides a sensitive means for detecting pollution related DNA adducts in aquatic organisms exposed to environmental carcinogens. However, the following factors need to be taken into consideration during the data interpretation: (1) species-specific, naturally occurring DNA modifications (or I-compounds) are found in aquatic organisms at levels which are highly season dependent; and (2) many aquatic organisms, particularly lower invertebrates, cannot form DNA adducts from common pollutants such as polycyclic aromatic hydrocarbons (PAHs). The level of natural adducts is especially high in lower invertebrates, such as sponges and sea-urchins during their reproductive phase in the spring time (March/April): in subsequent months adducts were either undetectable or present at only trace levels. These invertebrates do not metabolize PAHs such as benzo[a]pyrene but readily biotransform aromatic amines such as 2-acetylaminofluorene to DNA-reactive forms. Pollution-related DNA adducts have been found in fish living in highly polluted rivers and marine sites and in carp exposed to an artificial Diesel-2/crude oil slick. In certain fish (English sole, brown bullheads, etc.) living in polluted environments, the formation of pollution-related DNA adducts has been correlated with an increased incidence of tumours. It is concluded that, while DNA adducts detected in aquatic organisms can be used for biomonitoring and detecting pollutants, there are several confounding factors that should be taken into consideration before one attempts to determine the type and concentration of carcinogenic pollutants present in aquatic environments. PMID- 8225509 TI - DNA adduct detection: some applications in monitoring exposure to environmental genotoxic chemicals. AB - In the assessment of genotoxic risk factors in the environment, the measurement of DNA adducts in aquatic organisms and in plants may have considerable implications. Using 32P-postlabelling, we have detected DNA adducts in the liver of carp (Chondrostoma nasus) from the River Rhone (France), both downstream and upstream from a polychlorinated biphenyl incineration plant. Some of the DNA adducts were specific to downstream fish, suggesting a differential pattern of exposure. We have also detected DNA damage in needles in a declining spruce forest. We found that, in the declining forest, the amounts of DNA adducts increase in relation to the degree of damage to the needles whereas, in a healthy forest, the levels of DNA adducts were low. We have also found DNA adducts in the leaves of hops grown in fields where heptachlor residues persisted. PMID- 8225510 TI - Testing of quantitative parameters in the 32P-postlabelling method. AB - The 32P-postlabelling technique involves many steps that need to be carefully controlled in order to obtain a reliable quantitative determination of DNA adducts. We have studied several of the parameters involved in the DNA digestion procedures as well as those concerned in the phosphorylation reaction. Since adducts behave in very different ways in the labelling reaction, an individual protocol has to be worked out for each particular type of adduct. Quantitation is usually possible only if a synthesized standard of the adduct under investigation is run in parallel to the DNA samples throughout the whole procedure. PMID- 8225511 TI - 32P-postlabelling in studies of polycyclic aromatic hydrocarbon activation. AB - Polycyclic aromatic hydrocarbons (PAHs) undergo metabolic activation reactions to yield intermediates that react with DNA to form covalent adducts. PAH administration leads to the formation of various types of DNA adducts that may differ between species, strains and tissues due to differences in metabolic activation and repair. The structures of PAH-DNA adducts can be identified by three approaches: co-chromatography with synthetic mononucleotide adduct standards; examining the adducts resulting from metabolism of pathway intermediates; or chemically blocking metabolic activation at specific sites on the PAH. Administration of putative metabolic intermediates of a PAH leads to enhanced formation of DNA adducts resulting from further activation along that pathway. Conversely, chemically blocking a bond or position on a PAH prevents adducts arising from activation at that site. By comparing the DNA adduct spectra generated by metabolites, blocked forms, and parent PAH administration, the pathways important in the metabolic activation of the PAH in each tissue may be deduced. Partial identification of these adducts may also be made by co chromatography with the products of reactions between synthetic reactive intermediates and defined polydeoxynucleotides, and more thorough identification by using synthetic DNA adduct standards. These approaches have all been successfully applied to studies of PAH activation, and are reviewed here. PMID- 8225512 TI - 32P-postlabelling in studies of arylamine and nitroaromatic hydrocarbon activation and mutagenesis. AB - Carcinogenic arylamines and nitroaromatic hydrocarbons are chemicals that present occupational health hazards and share pathways of metabolic activation. The 32P postlabelled DNA adducts formed in Chinese hamster ovary (CHO) cells treated with metabolites from two pathways that are common to the activation of the nitroaromatic hydrocarbon 6-nitrochrysene (6-NC) and the arylamine 6 aminochrysene (6-AC) compared with the spectra of mutations induced at the CHO hprt locus by these were metabolites. 6-Nitrosochrysene (6-NOC), which is reduced by the cells to N-hydroxy-6-AC, formed adducts mainly with deoxyguanosine, but induced mutations primarily through base-pair substitution involving deoxyadenosine. In contrast, 6-AC 1,2-dihydrodiol produced DNA adducts and mutations that mainly involved deoxyguanosine residues. The two major activation pathways for 6-NC and 6-AC thus produce distinct adduct and mutation spectra in CHO cells, and these adduct and mutational spectra are different from those of other arylamines and nitroaromatic hydrocarbons that have been studied. PMID- 8225513 TI - Some safety procedures for handling 32P during postlabelling assays. AB - 32P is a high-energy (1.7 MeV) beta-emitter. Its handling is therefore subject to regulation and very strict control. During the postlabelling procedure, numerous steps involve exposure to 32P. The main risk from exposure is through irradiation, but direct accidental contamination can occur. The various manipulation steps (ATP synthesis, labelling, chromatography, quantification) have been analysed for their contribution to potential radiation exposure. Several measures have been taken in the IARC laboratories to minimize exposure of all personnel involved, including those who handle radioactive wastes, since most of the initial radioactivity is eventually discarded. The various steps to be taken for minimizing exposure, such as the training of personnel using fluorescent compounds instead of radioactivity, the use of protective screens and of equipment specially adapted for this work, are reviewed. PMID- 8225514 TI - Postlabelling methods for the detection of apurinic sites and radiation-induced DNA damage. AB - This paper reviews the 32P-postlabelling techniques that have been employed in the detection and quantitation of apurinic sites and DNA damage induced by UV and ionizing radiation. The two major approaches utilize different enzymes for DNA digestion--in one case, micrococcal nuclease and calf spleen phosphodiesterase, and in the other DNase I, snake venom phosphodiesterase and calf intestinal phosphatase. As a result, each technique detects different classes of lesions with limited overlap and can, therefore, be considered complementary. The enzyme dependence and other technical aspects, in particular problems of background resulting from undamaged DNA, and some of the applications with cellular DNA and naked DNA, are discussed. PMID- 8225515 TI - Anterior capsular support for posterior chamber intraocular lenses following vitreous loss in endocapsular surgery. AB - We used anterior capsular support for posterior chamber intraocular lenses (PC IOLs) in fourteen eyes with large posterior capsular ruptures. An endocapsular technique preserved the anterior capsule and facilitated implantation. With a median follow up of 8.5 months, all patients had a visual acuity of 6/9 or better. One lens was lost in the vitreous and one patient had a clinically significant cystoid macular edema. In the event of a posterior capsular rupture we suggest this technique as an alternative to anterior chamber or scleral fixated lenses. PMID- 8225516 TI - Recurrent pterygia--laser therapy: a preliminary report. AB - We report our experience with the use of Argon laser photocoagulation for the treatment of recurrent pterygia. In this study, 36 patients and 42 eyes with recurrent pterygia having undergone previous surgery one or more times, were subjected to laser therapy. The patients were followed up for a period of 9-12 months. The success rate was 92.8% when laser therapy was employed in recurrent pterygia which were excised once, while it was 64.2% in cases where excisions were done two or more times. Argon laser treatment is a safe and effective method of therapy for recurrent pterygia. PMID- 8225517 TI - The efficacy of acetone in the sterilisation of ophthalmic instruments. AB - Acetone has been considered a quick, effective and less expensive chemical sterilising agent and continues to be used by ophthalmic surgeons, at least in developing countries. Its utility however has been questioned recently. This study was designed to assess the efficacy of acetone against Pseudomonas aeruginosa, Bacillus subtilis and Aspergillus flavus present on ophthalmic surgical instruments (forceps, sutures). The instruments were contaminated by immersion in standard suspensions of the organisms and thereafter were either unwashed (group-I), washed and dried (group-II) or only washed (group-III) before immersion in acetone. The exposure to acetone was kept at 3, 10 and 20 minutes in each group. The results showed that acetone could eliminate Pseudomonas (vegetative bacteria) after 10 minutes exposure in unwashed group and 3 minutes exposure in washed groups. It was ineffective against spore bearing bacteria (B. subtilis) and fungus (Aspergillus flavus) even after 20 minutes of exposure. PMID- 8225518 TI - A genetic analysis of retinitis pigmentosa. AB - The data consists of sixty probands affected with Retinitis pigmentosa. Syndromic cases were found in five percent of the RP probands. Segregation analysis was carried out on proband sibship data. The ascertainment probability was estimated at 0.5517. Analysis of the data by parental mating types of proband sibships indicated the presence of dominant forms of RP (2.05%). Analysis of proband sibships indicated the presence of low risk families in the Normal x Normal matings (45%) and in the consanguineous matings (40%). The hypothesis of recessive inheritance could be confirmed only in multiplex sibships (p = 0.383 +/ 0.0793). Data on proband matings though incomplete conformed in general to autosomal recessive gene hypothesis. PMID- 8225519 TI - Malignant melanoma of the conjunctiva. PMID- 8225520 TI - Familial involvement in congenital fibrosis syndrome, a rare association with myopia. PMID- 8225521 TI - Changing patterns of infectious keratitis: overview of clinical and histopathologic features of keratitis due to acanthamoeba or atypical mycobacteria, and of infectious crystalline keratopathy. AB - Acanthamoeba keratitis, infectious crystalline keratopathy and atypical mycobacterial keratitis have recently emerged as important types of infectious keratitis. These corneal infections have been associated with contact lens wear and with corneal surgical procedures such as radial keratotomy and penetrating keratoplasty, and the clinical setting of each of these infections is important in alerting the clinician to the possible diagnosis. There have been improvements in rapid diagnostic techniques for such infections in the last several years. Treatment has also improved, but remains a difficult problem, especially for Acanthamoeba. An overview of recent developments in the clinical and histopathologic methods for diagnosis and treatment options of these three corneal infections is provided. PMID- 8225522 TI - Degsagittalisation of obliques: a case report of plagiocephaly with inferior oblique overaction. PMID- 8225523 TI - Plasmacytoma of orbit. PMID- 8225524 TI - Guidelines for the management of rhegmatogenous retinal detachment. AB - Current techniques of rhegmatogenous retinal detachment repair allow most detachments to be repaired successfully. The success of repair depends on a careful pre-operative examination and choice of an appropriate procedure. The surgery is usually tailored to individual needs. Improvements in surgical techniques coupled with a better understanding of the pathophysiology of rhegmatogenous retinal detachment continue to improve the anatomic and functional success of retinal detachment repair. PMID- 8225525 TI - Postexercise hypotension. Key features, mechanisms, and clinical significance. AB - Recent investigations have demonstrated that there is a sustained reduction in arterial blood pressure after a single bout of exercise, ie, postexercise hypotension (PEH). The purpose of this discussion is to integrate the available information on this topic and to review studies using sustained stimulation of somatic afferents in experimental rats as a model to study the role of somatic afferents in PEH. PEH occurs in response to several types of large-muscle dynamic exercise (ie, walking, running, leg cycling, and swimming) at submaximal intensities greater than 40% of peak aerobic capacity and exercise durations generally between 20 and 60 minutes. PEH is observed in both normotensive and hypertensive humans and in spontaneously hypertensive rats but is generally greater in magnitude in hypertensive subjects. The maximal exercise-induced reductions in systolic and diastolic arterial blood pressures have been on average 18 to 20 and 7 to 9 mm Hg, respectively, in hypertensive humans and 8 to 10 and 3 to 5 mm Hg, respectively, in normotensive humans. PEH has been reported to persist for 2 to 4 hours under laboratory conditions. Whether PEH is sustained for a prolonged period of time under free-living conditions remains controversial, although the results of one study indicate that PEH can persist for up to 13 hours. Possible mechanisms involved in mediating postexercise and poststimulation reductions in arterial blood pressure include decreased stroke volume and cardiac output; reductions in limb vascular resistance, total peripheral resistance, and muscle sympathetic nerve discharge; group III somatic afferent activation; altered baroreceptor reflex circulatory control; reduced vascular responsiveness to alpha-adrenergic receptor-mediated stimulation; and activation of endogenous opioid and serotonergic systems. It appears that the magnitude of PEH in hypertensive subjects is clinically significant; however, more investigation is required to determine if the duration is sufficient under real-life conditions to contribute to the reduction in blood pressure observed with chronic exercise conditioning. PMID- 8225526 TI - Arterial smooth muscle cell phenotype in stroke-prone spontaneously hypertensive rats. AB - The aim of this study was to determine the phenotype of smooth muscle cells in the arteries of chronically hypertensive animals and to analyze the effects of treatments known to increase the survival of the animal without a clear effect on its hypertensive state. Stroke-prone spontaneously hypertensive rats (SHRSP) kept on a 1% sodium drinking solution were untreated or treated with one of two diuretics, indapamide (3 mg/kg per day) or hydrochlorothiazide (20 mg/kg per day), from 6 to 13 weeks of age. Phenotype was characterized by the immunolabeling of arteries with antibodies raised against a cellular form (EIIIA) of fibronectin, alpha-smooth muscle actin, and nonmuscle myosin. We demonstrated that phenotypes of smooth muscle cells of the SHRSP differ from those found in Wistar-Kyoto rats. The difference in phenotype is specific for the vessel type: ie, an increased expression of nonmuscle myosin in the aorta and of both EIIIA fibronectin and nonmuscle myosin in the coronary arteries. The two diuretics (1) had no effect on blood pressure, (2) prevented or did not prevent the increase in medial thickness, and (3) prevented changes in both smooth muscle cell phenotype and ischemic tissular lesions. Taken together, the results suggest that in SHRSP the changes in the phenotype of smooth muscle cells and the thickness of arteries are unrelated events. We propose that the maintenance of the contractile phenotype of the arterial smooth muscle cells could be an essential parameter involved in the prevention of the deleterious consequences characteristic of a severe hypertensive state. PMID- 8225527 TI - ATP-sensitive potassium channels in the basilar artery during chronic hypertension. AB - We examined the hypothesis that dilatation of the basilar artery in response to activation of ATP-sensitive potassium channels is impaired in stroke-prone spontaneously hypertensive rats (SHRSP). Changes in basilar artery diameter in response to aprikalim, a direct activator of ATP-sensitive potassium channels, were measured in anesthetized SHRSP and normotensive Wistar-Kyoto (WKY) rats through a cranial window. Topical application of aprikalim increased basilar artery diameter in WKY rats. Glibenclamide, a selective inhibitor of ATP sensitive potassium channels, abolished aprikalim-induced vasodilatation. Thus, ATP-sensitive potassium channels are functional in the basilar artery of WKY rats in vivo. Aprikalim (10(-6) mol/L) dilated the basilar artery by 31 +/- 5% (mean +/- SEM) in WKY rats but only 5 +/- 1% in SHRSP. The concentration-response curve to aprikalim in SHRSP was significantly shifted to the right, but the response to the highest concentration of aprikalim (10(-5.5) mol/L) was similar in SHRSP and WKY rats. Vasodilatation in response to norepinephrine was also impaired in SHRSP. Dilator responses of the basilar artery to forskolin, a direct activator of adenylate cyclase, and nitroprusside, a direct activator of guanylate cyclase, were normal in SHRSP. The findings suggest that dilatation of the basilar artery in response to direct activation of ATP-sensitive potassium channels is impaired in SHRSP compared with WKY rats in vivo. PMID- 8225528 TI - Long-term low-dose angiotensin converting enzyme inhibitor treatment increases vascular cyclic guanosine 3',5'-monophosphate. AB - We investigated functional changes in aortic preparations of spontaneously hypertensive rats treated in utero and subsequently up to 20 weeks of age with the angiotensin converting enzyme (ACE) inhibitors ramipril (0.01 and 1 mg/kg per day) and perindopril (0.01 mg/kg per day). Early-onset treatment with the high dose of ramipril inhibited aortic ACE activity, prevented the development of hypertension, increased aortic vasodilator responses to acetylcholine (10(-8) to 10(-6) mol/L), decreased vasoconstrictor responses to norepinephrine (10(-8) mol/L), and increased aortic cyclic GMP content by 160%. Low-dose ramipril inhibited aortic ACE activity and attenuated the aortic vasoconstrictor response to norepinephrine but had no effect on blood pressure. Low-dose treatment with ramipril and perindopril resulted in a significant increase in aortic cyclic GMP content by 98% and 77%, respectively. Long-term coadministration of the bradykinin B2-receptor antagonist Hoe 140 abolished the ACE inhibitor-induced increase in aortic cyclic GMP. Our data demonstrate that long-term treatment with ACE inhibitors can alter vascular function of compliance vessels independently of the antihypertensive action. The increase in aortic cyclic GMP was due to bradykinin potentiating the action of the ACE inhibitors. PMID- 8225529 TI - Angiotensin II/prostaglandin I2 interactions in spontaneously hypertensive rats. AB - The purpose of this study was to compare the effects of prostaglandin I2 on several cardiovascular parameters and to compare the ability of prostaglandin I2 to modify angiotensin II-induced changes in these cardiovascular parameters in spontaneously hypertensive versus normotensive rats. Studies were conducted in adult, age-matched, indomethacin-, and captopril-pretreated spontaneously hypertensive and normotensive rats that had been prepared for assessment of arterial blood pressure, cardiac output (thermodilution), and renal and mesenteric blood flows (transit-time flow probes). In both normotensive and hypertensive rats, intravenous infusions of prostaglandin I2 (0.003, 0.03, 0.3, 1, 3, and 10 micrograms/kg per minute) dose-dependently reduced mean arterial blood pressure, total peripheral resistance, and mesenteric vascular resistance but not renal vascular resistance. Only minor differences were detected between normotensive versus hypertensive rats with regard to the effects of prostaglandin I2 on baseline cardiovascular parameters (ie, in the absence of angiotensin II). In both rat strains, an intravenous infusion of angiotensin II (300 ng/kg per minute) increased mean arterial blood pressure, total peripheral resistance, and renal and mesenteric vascular resistances, and these effects of angiotensin II were similar in the two strains in the absence of prostaglandin I2. In both strains, prostaglandin I2 inhibited angiotensin II-induced changes in mean arterial blood pressure, total peripheral resistance, and renal and mesenteric vascular resistances. However, in the renal, but not mesenteric, vasculature of hypertensive rats, the ability of prostaglandin I2 to attenuate angiotensin II induced vasoconstriction was strikingly reduced. These results indicate that although in general spontaneously hypertensive rats respond normally to prostaglandin I2, in the kidney of spontaneously hypertensive rats the ability of prostaglandin I2 to attenuate angiotensin II-induced vasoconstriction is reduced. This selective renal defect may relate to the pathogenesis of high blood pressure in this genetic model of hypertension. PMID- 8225530 TI - Angiotensin II increases norepinephrine release from atria by acting on angiotensin subtype 1 receptors. AB - Norepinephrine stores in electrically driven guinea pig isolated atria were loaded with [3H]norepinephrine, and norepinephrine release was deduced from the radioactivity efflux. Electrical field stimulation of sympathetic nerve endings was applied during the refractory period of atrial contractions. The stimulation induced release of norepinephrine was increased by angiotensin II (Ang II) (10( 8) to 10(-6) mol/L) in a concentration-dependent manner. The maximum observed effect was a 55% augmentation. The effects of 10(-7) and 10(-6) mol/L Ang II were abolished by 10(-6) and 10(-5) mol/L of the subtype 1 Ang II receptor antagonist losartan, respectively. Losartan by itself (10(-6) mol/L) caused a 14% reduction of norepinephrine release. The subtype 2 Ang II receptor ligand PD 123319 (1-[[4 (dimethylamino)-3-methylphenyl]methyl]-5-(diphenylacetyl)- 4,5,6,7-tetrahydro-1H imidazo[4,5-c]pyridine-6-carboxylic acid ditrifluoroacetate) in a concentration of 10(-4) mol/L had no detectable influence on transmitter release and did not antagonize the effect of Ang II. Angiotensin I (10(-6) and 10(-5) mol/L) increased norepinephrine release maximally by 23%. This effect was antagonized by 10(-5) mol/L losartan and did not appear in the presence of 10(-6) mol/L of the converting enzyme inhibitor ramiprilat. These results suggest that Ang II increases norepinephrine release by an activation of subtype 1 receptors, whereas angiotensin I is converted to Ang II to become effective. PMID- 8225531 TI - Cardiac adenylyl cyclase, beta-adrenergic receptors, and G proteins in salt sensitive hypertension. AB - The present study investigated whether high salt intake (8%) in Dahl salt sensitive and salt-resistant rats with and without hypertension produces a heterologous desensitization of cardiac adenylyl cyclase as observed in various types of hypertension and human heart failure. In membranes from Dahl salt sensitive rats on a high-salt diet (8%) basal, isoproterenol-, 5' guanylylimidodiphosphate-, and forskolin-stimulated adenylyl cyclase was reduced compared with the low-salt (0.4%) group and Dahl salt-resistant rats on either 0.4% or 8% sodium chloride. The activity of the catalyst was depressed, and the expression of the immunodetectable inhibitory G proteins Gi alpha was increased in Dahl salt-sensitive rats on 8% sodium chloride, whereas the density of beta adrenergic receptors and the activity of the stimulatory G protein Gs alpha reconstituted into Gs alpha-deficient S49 cyc- mouse lymphoma cell membranes were unchanged in any condition studied. We conclude that high salt intake in salt sensitive hypertensive Dahl rats produces hypertension, cardiac hypertrophy, and heterologous desensitization of cardiac adenylyl cyclase. The latter alteration is due to an increase of Gi alpha proteins and a depressed catalyst activity of adenylyl cyclase. The results demonstrate that heterologous adenylyl cyclase desensitization can precede the development of contractile dysfunction in later stages and can occur independently of changes in beta-adrenergic receptors. PMID- 8225532 TI - Natriuretic and hypotensive effect of adenosine-1 blockade in essential hypertension. AB - We studied the effects of a single dose (100 mg orally) and repeated administration (100 mg o.d. for 7 days) of FK453, a novel adenosine-1 receptor antagonist, on renal sodium handling and blood pressure in eight patients with essential hypertension. Within 60 minutes after administration of FK453, sodium excretion increased threefold. This occurred in the absence of a change in renal hemodynamics, assessed from inulin and para-aminohippurate clearance, and was accompanied by increased fractional excretion of lithium, phosphate, and uric acid and by increased excretion of calcium and magnesium. Maximal free water clearance data showed an increase in maximal urine flow and distal delivery term and a decrease in the diluting segment reabsorption term. FK453 also decreased blood pressure and increased heart rate, but this did not occur until about 3 hours after ingestion, that is, when the natriuresis was already over. The natriuretic effect of FK453 was short-lasting, and continued use of FK453 was in fact accompanied by some net sodium retention. Blood pressure on the seventh day before FK453 treatment was not different from blood pressure before administration of the first dose of FK453. Again an acute natriuretic response followed, although less than after the first dose. Changes in intrarenal sodium handling parameters, blood pressure, and heart rate were similar to those seen after the first dose. The natriuretic and hypotensive effects of FK453 indicate that adenosine-1 receptor activity plays a role in the regulation of blood pressure and renal sodium handling in patients with essential hypertension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225533 TI - Insulin reduces contraction and intracellular calcium concentration in vascular smooth muscle. AB - Resistance to insulin-induced glucose disposal is associated with hypertension, in accord with recent reports that insulin-induced vasodilation is impaired in men with resistance to insulin-induced glucose disposal. Nevertheless, the mechanism of insulin-induced vasodilation is not known. We wished to determine whether a physiological concentration of insulin inhibits agonist-induced contraction at the level of the individual vascular smooth muscle cell, and if so, how. Dispersed vascular smooth muscle cells from dog femoral artery were grown on collagen gels for 4 to 8 days. Contraction and intracellular Ca2+ concentration of individual cells were measured by photomicroscopy and fura 2 epifluorescence microscopy, respectively. Serotonin and angiotensin II contracted cells in a dose-dependent manner. Preincubation of cells for 20 minutes (short term) or 7 days (long-term) with insulin (40 microU/mL) inhibited serotonin- and angiotensin II-induced contractions by approximately 50%. Insulin (10 microU/mL) acutely inhibited serotonin-induced contraction by 34%. The maximal effect of high extracellular K(+)-induced contraction was not affected by short-term insulin exposure, but the ED50 for extracellular K(+)-induced contraction was increased from 7.6 +/- 2.5 to 16.0 +/- 3.9 mmol/L (P < .05). Short-term insulin exposure also attenuated the peak rise of the serotonin-induced intracellular Ca2+ transient and increased the rate constant for intracellular Ca2+ decline. Verapamil and ouabain completely blocked the attenuation of agonist-induced contraction by short-term insulin exposure, indicating the importance of voltage operated Ca2+ channels and the Na(+)-K+ pump for this effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225534 TI - Coarctation induces alterations in basement membranes in the cardiovascular system. AB - A coarctation hypertensive rat model was used to examine the effects of elevated blood pressure on basement membrane component synthesis by cardiac myocytes and aorta using immunohistochemistry and Northern blot analysis. Carotid arterial pressure increased immediately on coarctation, and left ventricular hypertrophy was maximal within 5 days. In immunohistochemical studies, fibronectin and laminin were increased and the basement membrane chondroitin sulfate proteoglycan decreased in both the subendothelial space and smooth muscle cell basement membranes of the aorta above the clip compared with controls, whereas only fibronectin was elevated in the aorta below the clip. No change in basement membrane staining intensity for the cardiac myocytes was observed. Alterations in steady-state mRNA levels for fibronectin and laminin in the aorta paralleled those observed by immunohistochemical analysis with regard to protein and tissue type affected as well as intensity of the changes. However, changes in mRNA levels (but not protein deposition) for perlecan and type IV collagen were also observed in aortas from hypertensive rats compared with controls. Increases in steady-state mRNA levels for all basement membrane components in the heart and vasculature peaked before maximal cardiac hypertrophy (5 days). These studies indicate that alterations in basement membrane component deposition in the hypertrophied vasculature occur at both transcriptional and translational levels and suggest that the cell attachment glycoproteins fibronectin and laminin may be important factors in the vascular response to elevated transmural pressure. PMID- 8225535 TI - Early interstitial changes in hypertension-induced renal injury. AB - To elucidate the mechanisms of hypertensive renal injury, we investigated the time course and extent of changes in matrix composition, as well as cell proliferation and infiltration in two-kidney, one clip rats. The nonclipped kidneys from hypertensive and sham-operated control rats (n = 5 to 10 in each group) were studied at 7, 14, 21, and 28 days after clipping. Systolic blood pressure was elevated by day 7 (154 +/- 3 versus 111 +/- 4 mm Hg in sham group, P < .001, n = 10 each). Hypertension resulted in an early expansion of the interstitial volume by 37%, whereas hypertensive vascular changes and glomerular injury did not become evident until day 21. Immunofluorescence studies revealed an early interstitial accumulation of collagens I, III, IV, V, VI, and fibronectin by day 7. In contrast, the glomeruli showed a mild to moderate increase in collagens I, III, IV, V, laminin, and fibronectin but not collagen VI later in the established phase of hypertension. Staining for proliferating cell nuclear antigen as a marker of cell replication was increased in tubular epithelial but not interstitial or glomerular cells. A progressive infiltration of macrophages (16 +/- 2 versus 9 +/- 1 ED1+ cells/mm2, P < .05, n = 6) and T lymphocytes (93 +/- 15 versus 74 +/- 7 CD4+ cells/mm2, n = 8) in the cortical interstitium had already occurred by day 7. On the other hand, only macrophages increased in number within the glomeruli. Thus, renovascular hypertension leads to an early tubular cell proliferation, mononuclear cell recruitment, and deposition of matrix proteins primarily within the interstitium. We conclude that the injury producing nephrosclerosis in this model extends far beyond the glomeruli. Both the tubules and the interstitium are actively involved and may be the more important initial sites of injury. PMID- 8225536 TI - 'White coat' hypertension. No harm for the heart. AB - The superiority of ambulatory blood pressure monitoring over casual blood pressure measurement for the prediction of target-organ damage is now well established, although the significance of "white coat" hypertension is still controversial. Is an office blood pressure measurement that is higher than the mean ambulatory value an added risk? Because left ventricular hypertrophy is a potent risk factor, the effect of hypertension on left ventricular mass merits attention. Left ventricular mass indexed for height was measured by M-mode echocardiography in 204 essential hypertensive patients (140 males, 64 females; mean age [SD], 50 [11] years). Blood pressure over 24 hours was monitored routinely with a noninvasive auscultatory device. Office blood pressure after 10 minutes of the subject in the supine position was measured by the same physician under the same conditions. The white coat effect was defined as the difference between the office and average daytime values of systolic and diastolic blood pressures. Patients were divided into four quartiles according to their office and daytime blood pressures. Two-way analysis of variance was used to assess the influence of the white coat effect at a given level of daytime blood pressure on left ventricular mass. Left ventricular mass increased significantly (P < .001) with daytime systolic blood pressure but was independent of white coat effect for both the whole population and the 143 untreated patients. We conclude that the white coat effect has no influence on left ventricular mass, favoring the view that white coat hypertension is a benign condition. PMID- 8225537 TI - Essential hypertension and cognitive function. The role of hyperinsulinemia. AB - The relation between hypertension and cognitive function is not well established. Therefore, we examined cognitive function in a random sample of 744 nondiabetic elderly inhabitants of Kuopio, East Finland. Five brief neuropsychological tests known to be sensitive to cognitive impairment due to dementia--the Mini-Mental State Examination (MMSE), the Trail Making Test (TMT), the Buschke Selective Reminding Test (BSR), Russell's Adaptation of the Visual Reproduction Test (HVR), and the Verbal Fluency Test (VFT)--were used to evaluate cognitive function. The performance of the hypertensive group (n = 378) was impaired in almost all test items compared with that of the normotensive group (n = 366), but the difference between these two groups was statistically significant in 5 of 19 test items only. Moreover, within the hypertensive group, hyperinsulinemic (fasting plasma insulin > 17.9 mU/L) hypertensive subjects (n = 57) scored worse than normoinsulinemic hypertensive subjects (n = 321) in 16 of 19 test items and worse than the normotensive subjects in the same 16 of 19 test items. The difference between the hyperinsulinemic hypertensive and normotensive groups was significant in 11 test items that reflected complex cognitive function such as calculation, language, semantic memory, and problem solving. This difference in neuropsychological tests among the three study groups (normotensive, normoinsulinemic hypertensive, and hyperinsulinemic hypertensive subjects) persisted after adjustment for fasting plasma glucose, age, sex, and education in 3 test items measuring calculation, copying, and semantic memory. Thus, essential hypertension in the elderly is associated with an impairment in complex cognitive function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225538 TI - Antihypertensive therapy and adaptive mechanisms in peripheral ischemia. AB - In the present experiments the effect of long-term peripheral ischemia on the capillary of two hind limb skeletal muscles was investigated in spontaneously hypertensive rats. Furthermore, the effect of antihypertensive therapy on changes in capillarity and on the previously observed hyperreactivity of the ischemic vascular bed to vasoconstrictors was investigated in perfused hind limbs of rats after long-term treatment with the angiotensin I converting enzyme inhibitors captopril (0.5 mg/kg.h) or zabiciprilate (0.025 mg/kg.h), the angiotensin II type 1 receptor antagonist losartan (0.625 mg/kg.h), or the calcium antagonist felodipine (0.042 or 0.42 mg/kg.h). Skeletal muscle ischemia in the left hind limb was induced by partial ligation of the left common iliac artery. Long-term (4 weeks) ischemia increased significantly the capillary-to-fiber ratio in the soleus muscle, composed predominantly of type I fibers in spontaneously hypertensive rats, of the ischemic hind limb, whereas capillarity in the contralateral muscle was not affected. Furthermore, capillarity in the gastrocnemius muscle (type II muscle fiber part) of both the ischemic and contralateral hind limb did not change. Long-term treatment with the angiotensin I converting enzyme inhibitors during ischemia abolished the increase in the capillary-to-fiber ratio in the soleus muscle, whereas a comparable antihypertensive dose of felodipine had no effect. Greater blood pressure reductions by both losartan and felodipine prevented increases in capillarization in skeletal muscle ischemia. With respect to vascular hyperreactivity during ischemia, only treatment with losartan normalized reactivity of the ischemic vascular bed to vasoconstrictors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225539 TI - Age-specific genetic effects for blood pressure. AB - Correlations between relatives were determined for systolic and diastolic blood pressure. The correlations decrease as age differences between relatives increase in a Norwegian sample with 43,751 parent-offspring pairs, 19,140 pairs of siblings, and 169 pairs of twins. A simple biometric model specifying only age specific genetic additive effects and environmental effects fitted well to correlations between cotwins, pairs of siblings, and parent-offspring dyads in subsets of relatives grouped by age differences. None of the environmental effects appeared to be due to environmental factors that are shared by family members. Models that excluded a parameter for the age-specific genetic influence did not fit the data. The results may partly explain what seems to be a discrepancy between relatively low parent-offspring correlations from previous nuclear family studies and high correlations from twin studies, especially in identical twins. PMID- 8225540 TI - Effect of stress on organ weight in rats. AB - The effect of 90 minute supine restraint or forced swim stress, on weights of various organs in sixty adult male albino rats was studied. One group of twenty rats served as control; second group of twenty was immobilized and third group of twenty was forced to swim in water at room temperature. The animals were studied for a period of 15 days. During this period, they were subjected to 90 minutes of stress everyday at the same time. Both the groups of stressed animals showed a statistically significant increase in weight of cerebrum, cerebellum, pituitary, thyroid, and adrenals. Testes and seminal vesicles showed no significant change in weight. There was an apparent increase in weight of liver after immobilization stress, and a statistically significant increase in its weight in forced swim group. Therefore, immobilization and forced swim stresses produce an increase in the weight of brain cerebrum and cerebellum, endocrine organs and liver. PMID- 8225541 TI - Magnesium and reserpine influence norepinephrine sensitivity of vas deferens in rats. AB - Reserpine induced supersensitivity to norepinephrine (NE) in rat vas deferens was sought by alteration of Mg++ and Ca++ concentration in incubation medium in the absence and presence of EDTA. Vas deferens incubated in Mg++ free and Mg++ excess media showed supersensitivity and subsensivity to NE respectively. Alterations in the sensitivity to NE produced by varying the concentrations of Mg++ were comparatively less. In the presence of EDTA, vas deferens obtained from reserpinized animals showed subsensitivity in normal and Mg++ excess media and supersensitivity in Mg++ free medium. In the presence of EDTA, reserpinized preparations showed slight supersensitivity in normal Mg++ medium, marked supersensitivity in Mg++ free and lesser subsensitivity in Mg++ excess medium. Probably EDTA by more effectively removing Mg++ from the membrane binding sites by chelation makes the membrane permeable to Ca++ leading to supersensitivity to NE (observed only in the presence of EDTA). These results suggest that the failure of reserpine to induce supersensitivity to NE in rat vas deferens may be due to an enhanced antagonism of Mg++ on Ca++ movements in this preparations due to the poor capacity of rat tissue to retain Ca++. PMID- 8225542 TI - Effect of lead on anorexia and body weight in albino rats. AB - Rats exposed to lead (lead acetate) in doses of 0.2 and 0.5 mg/ml in drinking water for a period of 90 days showed mild to moderate changes in food consumption compared to control group. Drug interactions in lead exposed rats with metoclopramide, atropine sulphate, propranolol, cyproheptadine and mepyramine maleate when administered intraperitoneally caused -30 to +30 percentage variation in food intake indicating the influence of adrenergic, serotonergic and cholinergic neurotransmitters with no change in mean body weight of lead treated rats. PMID- 8225543 TI - Does Aerua lanata have diuretic properties? AB - We have compared the diuretic activity of 200 ml of sucrose flavoured aqueous extract of the herb Aerua lanata with that of 200 ml sucrose flavoured weak infusion of tea leaves, and 200 ml of water, in 14 healthy volunteers under standardized conditions. Each volunteer acted as his or her own control in the three protocols, and chemical, physical and statistical analyses were performed "blind" by coding all urine and blood samples collected. A. lanata extract did not significantly increase urine flow, sodium excretion, potassium excretion, or urine and plasma osmolality as compared to an infusion of tea or plain water. PMID- 8225544 TI - Prediction equations for maximal voluntary ventilation in non-smoking normal subjects in Madras. AB - Maximal voluntary ventilation (MVV) was measured in 256 healthy non-smoking adults (132 males, 124 females) aged 15-63 years living in Madras. The mean MVV (+/- SD) in males was 126.7 +/- 31.9 and in females 77.7 +/- 16.4. Regression equations were derived for men and women for predicting maximal voluntary ventilation for adults in South India. MVV in South Indians were similar to those reported for other Indian subjects, but lower than those reported for caucasians. PMID- 8225545 TI - A computer programme for recording male sex behaviour in rats. AB - An accurate method of recording the frequencies of copulatory events, the latencies of initiation to copulation and the time spent in different behavioural categories is described. A microcomputer (IBM-PC) based data collection system for acquisition and analysis of male rodent sex behaviour has been developed. This software features ease of data entry and operation, using single key presses by assigning a preset code to each. Internal clock of the computer is made to function as a timer for accurate recording of latencies and intervals. A print out of the frequency or duration of data can be obtained either concurrently or after the completion of the experiment, as required. The least count of the technique is about 10(-4) min and this precludes its use for extremely rapidly changing behaviour. PMID- 8225546 TI - Reflex reversal of nostril dominance by application of pressure to the axilla by a crutch. AB - Effect of pressure application by a crutch to the axilla of the side of the dominant right nostril on the pattern of nostril dominance with emphasis on the time sequence of changes and recovery was studied in the young male Nigerians. The nostril dominance was assessed from the percentage of tidal volume flowing through the right and left nostril passages during expiratory phase (7). Application of pressure to right axilla for 20 min caused tidal volume flowing through left nostril to change from the control value of 20.3 +/- 15.8% (SD) to 74.1 +/- 10.4% while the rest of the tidal volume passed through the right nostril. The effect started in the first minute; air flow through both nostrils was equalized by 4 min; reversal of nostril dominance occurred thenceforth; and peak change was obtained by 17 min after application of pressure to the axilla. Removal of crutch could not lead to recovery. PMID- 8225547 TI - In vitro effect of alloxan on Na(+)-K(+)-ATPase and succinate dehydrogenase activities in brain and liver of mice. AB - The present study reports in vitro inhibition of the activities of enzymes Na(+) K(+)-ATPase and succinate dehydrogenase by alloxan in brain and liver homogenates of Swiss mice. The Vmax of both the enzymes was reduced in presence of alloxan without any substantial alteration in Km for substrate. Lineweaver Burk's plots showed higher 1/Vmax for alloxan treated samples and convergence of both slopes to intercept-1/Km. The observations pointed to non-competitive type inhibition of the enzymes by alloxan. This may be due to the modification of essential--SH groups present within/adjacent to substrate binding sites by alloxan. PMID- 8225548 TI - Effect of mianserin on blood glucose level in rabbits. AB - Blood glucose level was estimated in 18 h fasted albino rabbits following acute feeding of graded doses of mianserin. Mianserin (6.0 mg/kg) produced a gradually increasing hyperglycemic effect which became significant (P < 0.01) at 10 h and onwards. This appears to be due to increased turnover and release of noradrenaline by the drug. The same dose of mianserin also produced glucose intolerance during early hours probably by interfering with gastrin functions. PMID- 8225549 TI - Effect of anti-platelet therapy (aspirin + pentoxiphylline) on plasma lipids in patients of ischaemic stroke. AB - Twenty-one patients of ischaemic stroke were put on prolonged administration of antiplatelet drugs (aspirin 320 mg once daily with pentoxiphylline 400 mg thrice daily). The serum lipids along with other biochemical parameters were estimated before starting the treatment and after completion of 2 months of therapy. No significant changes were observed in any of the biochemical parameters including lipid profile except in serum high density lipoprotein (HDL) which increased significantly (< 0.05) after 2 months therapy. It is concluded that 2 months antiplatelet therapy has no adverse metabolic effect in patients of ischaemic stroke and the raised serum HDL may contribute to cerebral protective effect. PMID- 8225550 TI - Programme for drawing line graphs on IBM personal computers. AB - A simple program for drawing line graphs on IBM Personal Computers is described here. This program is written in Basic language and is user friendly. This program allows the operator to plot the line graphs with standard error of each of the observations. After plotting suitable legend can also be added at appropriate places in the graph. In the graphic mode a hard copy can be obtained from a dot matrix printer using print screen command. PMID- 8225551 TI - Zinc counteracts experimentally-induced cirrhotic changes in rats. AB - In the present study, cirrhosis was induced in rats by administration of carbon tetra chloride for 8 weeks. In these animals ZnSo4 (equivalent to 100 and 200 micrograms of zinc) was administered orally and liver function tests and plasma zinc (Zn) estimations were carried out after 2 and 4 week intervals. The results revealed that Zn supplement counteracts cirrhotic changes in liver. PMID- 8225552 TI - Brain cholesterol and phospholipid levels in cyproheptadine treated albino rats. AB - Effect of different doses (2.5 mg, 5 mg, 10 mg and 40 mg/kg), s.c., for 2 weeks, of cyproheptadine (CYP) on brain cholesterol and phospholipid of albino rats was investigated. Cyp. 2.5 mg/kg, showed increase in brain cholesterol and phospholipid contents whereas other doses caused a decrease in phospholipid level. PMID- 8225553 TI - Arginine vasopressin potentiates methyldopa induced hypotension in anesthetized Wistar rats. PMID- 8225554 TI - Karl landsteiner (1868-1943) PMID- 8225555 TI - Effects of mesencephalic lesion on testes, seminal vesicles and adrenal glands of albino rats. AB - The role of mesencephalic reticular formation on testicular structure, seminal vesicles, adrenal glands and kidneys of albino rats was investigated. Mesencephalic reticular formation lesions were produced using two epoxy-coated stainless steel electrodes. Electrolytic lesions resulted in atrophy of the testes, seminal vesicles and adrenal glands. The degree of atrophy was statistically significant in these organs (P < 0.01). However, there were no significant changes in the kidneys of lesioned animals. This study strongly suggests that the mesencephalic reticular formation influences the histomorphology of testes, seminal vesicles and adrenal glands. PMID- 8225556 TI - Differences in glycogen metabolism in astroglia-rich primary cultures and sorbitol-selected astroglial cultures derived from mouse brain. AB - Recently it has become possible by chemical selection using sorbitol instead of glucose in the culture medium to produce pure astroglial cultures from astroglia rich primary cultures from mouse brain. The glycogen-degrading enzyme glycogen phosphorylase in brain is localized in astrocytes and ependymal cells. In view of this fact it appeared necessary to study the influence of glucose and other hexoses on the glycogen metabolism in these cultures lacking the influence of other cell types in comparison to the astroglia-rich primary cultures containing several types of cells. The sorbitol-fed selected cultures and the glucose deprived astroglia-rich primary cultures contain less than 10% of the glycogen encountered in glucose-fed primary cultures. During incubation with glucose the glycogen content of the selected cultures and the glucose-deprived primary cultures increases by more than one order of magnitude. Nevertheless, not all cells are found to have accumulated glycogen. The time course of the replenishment of glycogen is similar in both types of culture, although maximal levels reached in the selected cultures are 3 times those in the astroglia-rich primary cultures. This difference might be explained by the fact that the ratio of the maximal activities of glycogen synthase and glycogen phosphorylase in selected cultures was found to be twice that in the unselected cultures. During glucose deprivation the glycogen content is reduced in both culture systems with half-maximal contents being reached at 15 min (primary culture) and 45 min (selected culture). Both types of culture can also utilize mannose for the synthesis of glycogen and the production of lactate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225557 TI - Migration of newly generated neurons upon ependymally derived radial guide cells in explant cultures of the adult songbird forebrain. AB - The adult songbird forebrain undergoes neuronal production throughout adulthood, with the production of new neurons in discrete regions of the neostriatal ventricular zone. Upon mitogenesis, these new neurons migrate into the subjacent brain parenchyma along radially directed guide fibers. In long-term ventricular zone explant cultures, derived from the higher vocal center of the adult canary, newly migratory neurons were found to associate preferentially with a characteristic substrate cell type. These small, parvonuclear substrate cells formed tightly packed epithelioid sheets, in which ciliated ependymal cells were common, as recognized by both live observation and electron microscopy. A subpopulation of these cells was immunostained by monoclonal antibody 3A7, which preferentially stains the guide fiber network of the adult avian brain. These 3A7+ cells included ependymal cells and bipolar radial cells, as well as morphologically defined astrocytes. As they matured in vitro, the 3A7+ bipolar radial cells extended long, unbranching fibers, which ultimately traversed the culture substrate. Like ependymal cells, they supported neuronal migration. These cells were likely homologous to radial guide cells in vivo. Thus, neuronal migration in adult avian forebrain culture occurred upon guide cells of ependymal derivation. PMID- 8225558 TI - N- and R-cadherin expression in the optic nerve of the chicken embryo. AB - Cadherins are a family of molecules mediating Ca(2+)-dependent cell-cell adhesion in various tissues. N- and R-cadherin are expressed in the chick embryonic CNS and differ in their expression pattern during development. Here we focus on the differential expression of N- and R-cadherin in the early optic nerve. N-cadherin is expressed by the retinal neurites growing through the optic nerve. R-cadherin is expressed by the early optic nerve glia, which derives from the optic stalk neuroepithelium and corresponds to an immature form of the type-1 astrocyte described in rat optic nerve. The close contact between the plasma membranes of the retinal neurites and the optic nerve glia is believed to be important in guiding retinal axons through the optic nerve. Using neuroblastoma cell lines transfected with R-cadherin, we demonstrate that the N-cadherin-positive retinal axons can use R-cadherin as a substrate for axon elongation. These results suggest that the R-cadherin expressed by the early optic nerve glia might provide a molecular substrate for the growth of N-cadherin-positive retinal axons through the optic nerve. PMID- 8225559 TI - Bergmann glia require continuous association with Purkinje cells for normal phenotype expression. AB - Bergmann glia (Bg) respond to the early postnatal Purkinje cell (Pc) death in Lurcher (Lc) mutant mouse cerebellum by down-regulating expression of the enzyme glycerol-3-phosphate dehydrogenase (GPDH). To determine whether glial GPDH expression requires the continued presence of Pcs in adults, we used single intracerebellar injections of kainic acid to kill Pcs in wild-type mice from 7 weeks to 11 months old. Bg at all ages tested responded to Pc loss by down regulating GPDH expression. To learn whether a high level of GPDH could be reinduced following down-regulation in Lc Bg, we grafted wild-type fetal Pcs into Lc cerebella. The influence of grafted Pcs on GPDH expression is host-age and implant-position dependent. Only Pcs implanted into hosts less than 6 weeks old were later found to be associated with GPDH-positive Bg. Grafted Pcs that migrated into the anterior folia of young hosts were more likely to be associated with GPDH-positive Bg than Pcs migrating to other positions. EM analysis showed that Bg ensheathment of grafted Pcs is thinner and more discontinuous, but qualitatively similar to normal. The results suggest that the interaction between host Bg and grafted Pcs can sustain elevated GPDH expression in Bg that have not yet down-regulated, but is not adequate to reinduce expression in those cells that have. PMID- 8225560 TI - Cell type-specific regulation of major histocompatibility complex (MHC) class I gene expression in astrocytes, oligodendrocytes, and neurons. AB - Mechanisms of major histocompatibility complex (MHC) class I gene regulation in cells of the CNS have been studied in vitro. Astrocytes in primary cultures, but neither oligodendrocytes nor neurons, constitutively expressed cell surface MHC class I molecules. Interferon-gamma (IFN-gamma) treatment led to induction of MHC class I expression in astrocytes and oligodendrocytes but not in neurons. The conserved upstream sequence containing the juxtaposed nuclear factor (NF)-kappa B like region I and IFN-response consensus sequence (ICS) constitutively enhanced MHC class I gene promoter activity in astrocytes, but not in oligodendrocytes or in neurons. Nuclear extracts from astrocytes, but not from oligodendrocytes and neurons, had a binding activity specific for the NF-kappa B-like region I sequence, indicating that constitutive expression of MHC class I genes is governed by the upstream region I enhancer and its binding factor. IFN-gamma treatment led to induction of MHC class I promoter activity in astrocytes and oligodendrocytes, but not in neurons. In accordance with this observation, a nuclear factor that binds to the ICS was induced in astrocytes and oligodendrocytes but not in neurons following IFN-gamma treatment. This study illustrates cell type-specific regulation of MHC class I genes in the CNS that correlates with the expression of DNA binding factors relevant to MHC class I gene transcription. PMID- 8225561 TI - Combination of a dual 5-lipoxygenase/cyclooxygenase inhibitor with a glucocorticoid results in synergistic topical antiinflammatory activity without inducing skin atrophy. AB - Synthetic glucocorticosteroids are widely used in clinical dermatology in the treatment of a number of inflammatory skin disorders. However, cutaneous side effects such as induction of skin atrophy are among the factors that limit the chronic use of this class of agents. We have found that a combination of a dual 5 lipoxygenase/cyclooxygenase inhibitor (9-phenylnonanohydroxamic acid, BMY 30094) and a glucocorticosteroid (hydrocortisone valerate) has synergistic antiinflammatory activity. Topical application of a 0.2% solution of hydrocortisone valerate alone inhibited phorbol ester-induced mouse skin inflammation by 92%. A 1% solution of BMY 30094 alone or a low dose of hydrocortisone valerate (0.005%) had no significant effect on the inflammatory reaction. A combination of 1% BMY 30094 and 0.005% hydrocortisone valerate inhibited the inflammation by 76%. This latter combination produced no signs of skin atrophy in rats after 28 days of application. In contrast, a 0.1% dose of hydrocortisone valerate produced significant skin atrophy. These findings suggest that it is possible to reduce the skin atrophy potential of glucocorticoids while maintaining the antiinflammatory activity using a novel drug combination. PMID- 8225562 TI - Elevated levels of xanthine oxidase in serum of patients with inflammatory and autoimmune rheumatic diseases. AB - Sera of patients with various inflammatory and autoimmune rheumatic diseases were screened for the presence of xanthine oxidase (XOD) and compared to sera from healthy donors and patients with nonrheumatic diseases including AIDS, internal diseases, and different carcinomas. Up to 50-fold higher levels of XOD were detected in rheumatic sera (P < 0.001). In addition, serum sulfhydryls (SH) were determined as sensitive markers of oxidative stress. The SH status in rheumatic patients was diminished by 45-75% (P < 0.001) and inversely correlated to the concentration of serum XOD (R = 0.73), suggesting a causal interrelation. The depletion of serum sulfhydryls by the oxyradical-producing XOD/acetaldehyde system was mimicked successfully ex vivo in human serum from healthy donors. Cortisone treatment of patients suffering from systemic lupus erythematosus and rheumatoid arthritis impressively normalized elevated XOD concentrations in rheumatic sera to those of healthy controls. The participation of xanthine oxidase in the depletion of serum antioxidants in rheumatic patients is discussed in the light of substrate availability and Km values. PMID- 8225563 TI - Effect of prostaglandins and superoxide dismutase administration on oxygen free radical production in experimental acute pancreatitis. AB - Oxygen free radicals and prostaglandins are implicated in the pathophysiology of acute pancreatitis, although their mechanisms of action remain unclear. We have studied the effect of administration of exogenous 16,16-dimethyl prostaglandin E2 and superoxide dismutase on oxygen free radical production in acute pancreatitis. For this purpose, five experimental rat groups were studied: group I, control; group II, sodium taurocholate-induced acute pancreatitis; group III, same as group II but with previous administration of 16,16-dimethyl prostaglandin E2; group IV, same as group II but with previous administration of indomethacin; and group V, same as group II but with previous administration of superoxide dismutase. In sodium taurocholate-treated rats, xanthine dehydrogenase is completely converted to xanthine oxidase within the first 5 min with subsequent oxygen free radical production while in 16,16-dimethyl prostaglandin E2-treated rats this enzyme transformation does not occur. In the superoxide dismutase treated group xanthine oxidase activation is partially prevented. These data suggest that xanthine oxidase is the main source of oxygen free radicals, which contribute to extending the cellular damage in sodium taurocholate-induced acute pancreatitis. PMID- 8225564 TI - Selective inhibition of group II phospholipase A2 by quercetin. AB - The influence of quercetin, chlorpromazine, aristolochic acid, and indomethacin on group I phospholipase A2 (PLA2) from porcine pancreas and on group II PLA2 from Vipera russelli was compared. Quercetin and chlorpromazine were found to inhibit PLA2 activity in lower concentrations (< 100 microM), while aristolochic acid and indomethacin were inhibitory only in higher concentrations (> 100 microM). The order of potency against Vipera PLA2 was: quercetin > chlorpromazine > aristolochic acid > indomethacin, while the order of potency against pancreatic PLA2 was: chlorpromazine > aristolochic acid > indomethacin >> quercetin. Thus, quercetin was a potent inhibitor towards group II PLA2 (IC50 = 2 microM), but a very weak inhibitor against group I PLA2, with maximum 30% inhibition. Aristolochic acid and indomethacin were three to four times more potent towards group II PLA2 than towards group I PLA2, while chlorpromazine was equally potent towards the two PLA2 types. Quercetin and chlorpromazine were also tested against two PLA2 fractions purified from the plasma of septic shock patients; chlorpromazine was then equally potent towards the two PLA2 fractions, whereas quercetin was a potent inhibitor of only one of the two PLA2 fractions (IC50 = 4 microM). Together, these results indicate that (1) different PLA2 inhibitors have different potency depending on which type of PLA2 they are used against, (2) quercetin selectively inhibits group II PLA2 and may therefore be used to discriminate between different PLA2 forms in biological materials, and (3) both PLA2 of group I and group II are present in septic shock plasma. PMID- 8225565 TI - Production of cytokines and PGE2 and cytotoxicity of stimulated bone marrow macrophages after thermal injury and cytotoxicity of stimulated U-937 macrophages. AB - Bone marrow-derived macrophages from normal and burned rats were cultured for one and four days in the presence of LPS, PHA, or opsonized zymosan as activators, and the supernatants were assayed for the inflammatory mediators TNF, IL-6, and PGE2 and the cells assayed for cytotoxicity. The macrophages responded differently to the various stimuli regarding cytotoxicity and the production of mediators, perhaps implicating the complement receptor CR1 in TNF production and the LPS receptor CD14 or the PHA lectin receptor in IL-6 and PGE2 production and for cytotoxicity. The response of the cells also depended on culture time and postburn time; in addition, macrophages from burned and unburned animals responded differently, depending on postburn day and the type of stimulus. TNF production was generally higher for one-day compared to four-day cultures (i.e., TNF was disappearing in the cultures), but IL-6 and PGE2 production was greater in four-day cultures. The results of this study suggest that thermal injury can contribute to the development of inflammatory and cytotoxic macrophages from bone marrow progenitor cells. PMID- 8225566 TI - Reactivity of an active center analog of Cu2Zn2superoxide dismutase in murine model of acute and chronic inflammation. AB - The antiinflammatory efficacy of CuPu(Py)2 ([[N,N'-bis(2-pyridylmethylene)-1,4 butanediamine] (N,N',N'',N''')]-Cu2+), a serum stable active center analog of Cu2Zn2superoxide dismutase (SOD), was tested in vitro and in vivo in male Wistar rats suffering from potassium peroxochromate-induced inflammation. Parameters including 99mTc gamma-scintigraphic imaging, the arthritis score, the plasma superoxide dismutase activity, the inhibition of plasma sulfhydryl depletion as well as mitogenic and phagocytic responses were used to quantify the disease activity. All parameters improved impressively during the treatment with CuPu(Py)2 and resembled those of healthy animals after 21 days. The arthritis score was inhibited by 80% (P > 0.001) and the plasma SOD activity enhanced by 380% (P > 0.001). The depletion of plasma sulfhydryls and the leukocytic responses to concanavalin A, tetradecanoylphorbolacetate, and lipopolysaccharide were significantly reduced (P > 0.001) and correlated well with the arthritis score. The collapse of antioxidant defenses in human plasma as well as the depolymerization of hyaluronic acid was mimicked in vitro and successfully inhibited by CuPu(Py)2. Oxidant-induced injury of plasma components during the aqueous decay of potassium peroxochromate were demonstrated to activate the oxidative burst of phagocytes in human blood. The role of impaired pro- and antioxidant balances in the etiology of inflammatory and autoimmune rheumatic diseases is discussed. PMID- 8225567 TI - Effects of serine protease inhibitor, TAME, on IL-1 beta in LPS-stimulated human monocytes: relationship between synthesis and release of a 33-kDa precursor and the 17-kDa biologically active species. AB - LPS stimulation of human monocytes in vitro induced release of the 17-kDa mature IL-1 beta (mIL-1 beta) but did not result in release of precursor IL-1 beta (pIL 1 beta). In contrast, the presence of a serine protease inhibitor, N alpha-(p toluene sulfonyl)-L-arginine methyl ester (TAME; 10 mM) for 6 or 18 h was associated with the LPS-stimulated release of the 33-kDa pIL-1 beta as well. These effects were initially discerned from observations that the fraction of the total IL-1 beta produced (as detected by ELISA) that was released from monocytes increased in the presence of TAME, and immunoblot assays confirmed that this fraction was predominantly 33-kDa IL-1 beta. A global decrease in monocyte protein synthesis was also observed after prolonged (18-h) exposure to TAME and was associated with a decrease in IL-1 beta synthesis, predominantly affecting 31 kDa pIL-1 beta, and a dose-dependent inhibition of TNF-alpha production. Parallel examination of lactate dehydrogenase (LDH) release indicated that pIL-1 beta release was unrelated to cell lysis. These results demonstrate that TAME inhibitable serine proteases are probably involved in the production and eventual proteolysis of the 33-kDa pIL-1 beta in situ but are probably not mechanistically related to either maturation of the IL-1 beta molecule or signaling of IL-1 beta release. IL-1 beta release appears to be dependent on the amount of total IL-1 beta synthesized. Serine proteolysis may constitute a degradative pathway for excess precursor, which, if interfered with, could result in release of the higher-molecular-weight forms of IL-1 beta. PMID- 8225568 TI - Shigella subversion of the cellular cytoskeleton: a strategy for epithelial colonization. PMID- 8225569 TI - Antigenic stability of Borrelia burgdorferi during chronic infections of immunocompetent mice. AB - Mice were actively immunized by intradermal inoculation with 10(4) cloned Borrelia burgdorferi bacteria and then cured of the B. burgdorferi infection with an antibiotic after 90 days. They were resistant to intradermal 10(2)- or 10(4) bacterium challenge infection with either the original cloned B. burgdorferi or B. burgdorferi isolated from each punch biopsies at 90 days of infection (prior to antibiotic treatment), including autologous B. burgdorferi isolates. In contrast, sham-infected (nonimmune) mice were susceptible to challenge infection with both early and late B. burgdorferi isolates. Since there was a potential for in vitro modification of the spirochetes during the 2-week culture period which would obscure results, an alternate means of challenge infection, using tissue transplants, was implemented. By using the same approach, mice were immunized by infection, treated with antibiotics, but challenged by subcutaneous transplantation of ear skin pieces biopsied and frozen prior to antibiotic treatment. Mice were infected for 15, 90, or 180 days before biopsy and antibiotic treatment and then transplant challenged with autologous infected tissue. Sham-immunized mice received infected tissue, and immune mice received uninfected tissue as controls. Mice infected for only 15 days, but not mice infected for 90 or 180 days, could be reinfected by autografts, whereas nonimmune mice became infected with tissues collected at each of these intervals and immune mice transplanted with normal skin were uninfected. These results indicate that immunity to B. burgdorferi is effective against the original inoculum, late isolates of the spirochete, or infected tissues collected at intervals of up to 180 days, suggesting that there is no significant antigenic change in B. burgdorferi during chronic infection. PMID- 8225570 TI - Characterization of Bartonella bacilliformis flagella and effect of antiflagellin antibodies on invasion of human erythrocytes. AB - Bartonella bacilliformis is the etiologic agent of Oroya fever in humans. Flagellum-mediated motility has been postulated as a major virulence factor for invasion of host cells. To address this hypothesis, we purified and characterized flagella from strain KC584 and then assessed their role in human erythrocyte association and invasion. Electron microscopy of the flagellar preparation showed a high concentration of filaments with a mean wavelength of 800 nm. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis, and KBr density gradient centrifugation indicated that the flagellar filament is composed of a polypeptide of 42 kDa. The flagellin is partially (ca. 50%) resistant to treatment with trypsin. The first 17 amino acid residues of the N terminus of the mature flagellin protein are GAAILTNDNAMDALQDL and show approximately 46% sequence identity to the residues of the N termini of two Caulobacter crescentus flagellin proteins. A monospecific polyclonal antibodies to the flagellin protein was generated, and its specificity was verified by both immunoblot and immunogold analyses. Human erythrocyte invasion assays performed with bartonellae exposed to the antiflagellin antiserum showed a significant decrease in bacterial association with and invasion of human erythrocytes in comparison with that in bartonellae exposed to preimmune rabbit serum or phosphate-buffered saline (PBS) controls. These results suggest that flagella are an important component in the invasiveness of B. bacilliformis. PMID- 8225571 TI - Staphylococcal alpha-toxin kills human keratinocytes by permeabilizing the plasma membrane for monovalent ions. AB - Incubation of human keratinocytes with nanomolar concentrations of Staphylococcus aureus alpha-toxin leads to irreversible depletion of cellular ATP. The toxin forms hexamers in the target cell membranes, and rapid transmembrane flux of K+, Na+, and 86Rb+ is observed. Unexpectedly, pores formed in keratinocytes through application of low but lethal doses of alpha-toxin appeared to be considerably smaller than those formed in erythrocyte membranes. They permitted neither rapid influx of Ca2+ or propidium iodide, nor efflux of carboxyfluorescein. Larger pores allowing flux of all three markers did form when the toxin was applied at high concentrations. Flux of monovalent ions and reduction in cellular ATP levels evoked by low toxin doses correlated temporally with a fall in oxygen consumption, which was interpreted to reflect breakdown of mitochondrial respiration. The lethal event could not be thwarted by manipulating the extracellular K+ or Ca2+ concentrations. Realization that alpha-toxin may form very small pores in nucleated cells is important for future research on cellular toxin effects and membrane repair processes. PMID- 8225572 TI - Involvement of tumor necrosis factor alpha in intracellular multiplication of Legionella pneumophila in human monocytes. AB - We investigated the role of tumor necrosis factor alpha (TNF-alpha) in human peripheral monocytes infected with Legionella pneumophila in vitro. Exogenous TNF alpha significantly inhibited the intracellular multiplication of the bacterium. This effect was concentration and time dependent and was abrogated by anti-TNF antibodies. TNF-alpha levels in the culture supernatants were low but were enhanced by the addition of gamma interferon. When monocytes were cultured and infected in the presence of pentoxyphilline, a potent inhibitor of TNF-alpha synthesis, the intracellular bacterial growth was enhanced. The effect of pentoxyphilline was concentration and time dependent and was due to the inhibition of TNF-alpha production, as shown by Northern (RNA) blot hybridization of total RNA. In addition, the pentoxyphilline partially abolished the inhibitory effect of gamma interferon on bacterial intracellular multiplication. These results suggest that gamma interferon inhibits, at least partially, the intracellular multiplication of L. pneumophila by enhancing TNF-alpha synthesis. PMID- 8225573 TI - Immunity after treatment of human schistosomiasis: association between cellular responses and resistance to reinfection. AB - Previous studies have demonstrated the development of an age-dependent resistance to reinfection after chemotherapeutic cure of the helminthic parasite Schistosoma mansoni. Here we report on a longitudinal investigation of cell-mediated responses in infected individuals before and after treatment which was designed to outline those parameters important in mediating a protective response. A well defined study group of 89 individuals with an age range of 9 to 35 years was selected from an area of high S. mansoni transmission in the Machakos district of Kenya. Peripheral blood mononuclear cell proliferation and cytokine production (interleukin-2 [IL-2], gamma interferon IL-5, IL-4, and tumor necrosis factor) in response to different crude life cycle-stage antigens of S. mansoni were assessed longitudinally in vitro before, 3 months after, and 1 year after treatment. Detailed statistical analyses of the results from this study have indicated a clear negative association between the proliferative responses to adult- and schistosomulum-stage antigens and subsequent reinfection intensity in older individuals (14 to 35 years) which was not present in the younger individuals (9 to 13 years). This association was significant even after the effects of age, sex, and exposure had been accounted for in multiple regression analyses. Cytokines were detected predominantly in response to adult worm and egg antigen extracts. An inverse association between the two cytokines gamma interferon and IL-5 was detected in response to all antigens at the three time points investigated, indicating cross-regulation in the production of these two mediators. Differences in antigen-specific cytokine levels between the two age groups were detected, with significantly higher IL-5 levels detected in the older (more resistant) age group. An inverse correlation between this cytokine and reinfection was detected but could not be dissociated from the effects of age and exposure in multiple regression analysis. PMID- 8225574 TI - Identification of the surface component of Streptococcus defectivus that mediates extracellular matrix adherence. AB - Bacterial attachment to host tissue is considered to be a crucial primary step in pathogen infection. Previous studies have shown that Streptococcus defectivus adheres specifically to cell-secreted extracellular matrix (ECM). Though generally not exposed in vivo, this host tissue is exposed at endothelial cell junctions and sites of tissue injury. In this report, we identify a ca. 200-kDa surface protein of S. defectivus involved in ECM adherence. Nitrous acid-derived mutant strains that were unable to bind ECM and which failed to adsorb adhesin specific antibody from polyclonal inhibitory sera were isolated. A surface protein (ca. 200 kDa) was absent from ECM-nonadherent mutants, indicating its involvement in ECM attachment. Additionally, affinity-purified antibody to the ca. 200-kDa protein inhibited whole-cell S. defectivus ECM attachment, whereas antibody to the same region of the nonadherent mutant cell wall-associated protein profile did not. Furthermore, solubilized cell wall-associated protein extracts of parent but not mutant strains bound ECM, confirming the significance of this protein in ECM adherence. Therefore, we propose that the ca. 200-kDa protein is the major S. defectivus surface component that mediates the ECM attachment of these organisms. PMID- 8225575 TI - Molecular analysis of the leukotoxin determinants from Pasteurella haemolytica serotypes 1 to 16. AB - All sixteen serotypes of Pasteurella haemolytica were shown to produce a leukotoxin protein which is immunologically related to the well-characterized serotype 1 leukotoxin. All of the leukotoxins were weakly hemolytic and were able to bind to BL-3 target cells. The leukotoxin determinants were characterized by Southern blot hybridization by use of the previously cloned serotype 1 determinant as the probe, and a number of distinct classes were identified. The leukotoxin determinants from serotypes 2, 3, and 11 were cloned. Nucleotide sequence analysis of the lktC and lktA genes of the serotype 3 and 11 determinants revealed nucleotide substitutions throughout the coding sequences. A comparison of the lktC and lktA genes and deduced proteins of serotypes 1, 3, and 11 showed that they are highly homologous. PMID- 8225576 TI - Correlation between vacuolating cytotoxin production by Helicobacter pylori isolates in vitro and in vivo. AB - Approximately 50 to 60% of Helicobacter pylori isolates produce a vacuolating cytotoxin in vitro. To assess cytotoxin production in vivo, we sought to determine whether infection with a Tox+ H. pylori strain is associated with the presence of serum antitoxin antibodies. H. pylori isolates and serum samples were obtained from 30 patients, and serum samples were obtained from 20 uninfected patients as controls. Sera were tested by enzyme-linked immunosorbent assay for reactivity with the purified 87-kDa vacuolating cytotoxin, and the 30 H. pylori isolates were tested for vacuolating cytotoxin production. Supernatants from 14 (47%) of the 30 H. pylori isolates induced vacuolation of HeLa cells. Sera from the 30 H. pylori-infected patients reacted with the purified 87-kDa cytotoxin to a greater extent than sera from the uninfected controls for both immunoglobulin G (IgG) and IgA classes (P = 0.0004 and P < 0.0001, respectively). Serum IgG and IgA responses to the purified 87-kDa cytotoxin were higher among the 14 patients infected with Tox+ strains than among the 16 patients infected with Tox- strains (mean optical densities +/- standard errors of the means of 0.603 +/- 0.11 versus 0.234 +/- 0.07 [P = 0.005] and 0.644 +/- 0.12 versus 0.341 +/- 0.08 [P = 0.04] for IgG and IgA, respectively). Infection with a Tox+ strain compared with a Tox- strain was associated with increased antral polymorphonuclear leukocyte inflammation scores (P = 0.04). These data indicate that cytotoxin production by H. pylori isolates in vitro correlates with cytotoxin production in vivo and that infection with Tox+ H. pylori isolates may be associated with increased antral mucosal polymorphonuclear leukocyte infiltration. PMID- 8225577 TI - Effects of multiplicity of infection, bacterial protein synthesis, and growth phase on adhesion to and invasion of human cell lines by Salmonella typhimurium. AB - Monolayers of intestine 407 (Int-407) cells were infected with the virulent Salmonella typhimurium strain C52, and the adhesion to and invasion of these cells were studied. The effects of the multiplicity of infection and growth phase of the bacteria (logarithmic versus stationary) on the interaction with eukaryotic cells were investigated. In contrast to other reports, we found no differences in the adhesive and invasive capacities of bacteria derived from logarithmic- or stationary-phase cultures. Invasion by S. typhimurium required bacterial protein synthesis and live Int-407 cells. Bacteria adhered equally well to dead or live Int-407 cells, which indicates that adhesion does not require metabolically active cells. Adhesion of S. typhimurium followed saturation kinetics, with a maximum of 10 adhesive bacteria per cell. This indicates that there is a limited number of bacterial adhesion sites (receptors) available on the surface of the host cell. Killed and live bacteria adhered equally well and competed with each other for cellular adhesion sites. This and adhesion experiments performed in the presence of chloramphenicol showed that bacterial protein synthesis is not required for adhesion. The general validity of the results obtained with S. typhimurium C52 was confirmed by comparing the invasion and adhesion data with those of the frequently used SL1344 and SR11 strains. In addition, we assayed the adhesion and invasion of S. typhimurium C52, SL1344, and SR11 and 27 S. typhimurium field isolates with Int-407, HeLa, and HEp-2 cells. PMID- 8225578 TI - Inactivation of the Streptococcus mutans wall-associated protein A gene (wapA) results in a decrease in sucrose-dependent adherence and aggregation. AB - A 0.8-kb HindIII-BamHI internal fragment of the Streptococcus mutans wall associated protein A gene (wapA) was ligated to the 5.1-kb HindIII-BamHI fragment of the chimeric Streptococcus-Escherichia coli plasmid pVA891 (Emr Cmr). The resulting construct was used to transform S. mutans GS-5, and erythromycin resistant mutants were isolated and analyzed. Directed mutagenesis of the wapA gene by plasmid insertion through homologous recombination was demonstrated by Southern blot hybridization with the wapA and pVA891 probes. Stable mutants were obtained, and the alteration of the wapA gene by insertional inactivation was associated with a significant decrease in S. mutans sucrose-dependent aggregation and binding to smooth surfaces. Thus, WapA may play an important role in the colonization of the tooth surface by S. mutans and in the buildup of dental plaque. These findings provided an explanation for previous studies which indicated that WapA was effective in the prevention of dental caries in animal models. Thus, the use of recombinant WapA in the preparation of a safe and effective human dental vaccine should be investigated further. PMID- 8225579 TI - Identification of coccoid Escherichia coli BJ4 cells in the large intestine of streptomycin-treated mice. AB - Escherichia coli BJ4, a rat isolate, was used to examine the growth and differentiation of the microorganism in its natural habitat, the intestine. Growth of E. coli BJ4 in the large intestine of streptomycin-treated mice was compared with its growth in laboratory media. By a number of methods, it was shown that E. coli BJ4 differentiates, during growth in the intestine, into two distinct populations, one that has the characteristics of the laboratory-grown strain and one that appears as a coccoid cell. Furthermore, it was shown that there is a natural selection for the coccoid-type cell in the intestine, while in laboratory media growth of rod-shaped E. coli BJ4 is enhanced. PMID- 8225580 TI - In vivo induction of apoptosis (programmed cell death) in mouse thymus by administration of lipopolysaccharide. AB - In vivo administration of bacterial lipopolysaccharide to mice induced DNA fragmentation in the thymus. Fragmented DNA was confirmed by agarose gel electrophoresis and laser flow cytometry. DNA fragmentation was predominantly detected in the thymus of young mice, while it was undetectable in the spleen, bone marrow, and lymph nodes. DNA fragmentation in the thymus was roughly dependent on the dose of lipopolysaccharide injected and reached the peak about 18 h after the injection. The addition of lipopolysaccharide to in vitro cultures of thymocytes did not cause DNA fragmentation, suggesting that lipopolysaccharide was unable to induce apoptosis of thymocytes directly. The injection of lipopolysaccharide induced no significant DNA fragmentation in adrenalectomized mice. The injection of anti-tumor necrosis factor alpha antibody together with lipopolysaccharide partially inhibited the appearance of DNA fragmentation in the thymus. On the basis of the fact that DNA fragmentation is one of the characteristics typical in apoptotic cell death, it was suggested that lipopolysaccharide could induce apoptosis in the mouse thymus in vivo. This apoptosis in the thymus might be mediated mainly by the adrenal hormones, but it is likely that tumor necrosis factor alpha might also participate in it. PMID- 8225581 TI - Transferrin and lactoferrin undergo proteolytic cleavage in the Pseudomonas aeruginosa-infected lungs of patients with cystic fibrosis. AB - Bacterium- and neutrophil-derived proteases have been suggested to contribute to tissue injury at sites of Pseudomonas aeruginosa infection. Pseudomonas elastase cleavage of transferrin enhances in vitro iron removal from this protein by the P. aeruginosa siderophore pyoverdin. This cleavage also generates new iron chelates which, in contrast to iron bound to transferrin, are able to catalyze formation of the highly cytotoxic hydroxyl radical from neutrophil-derived superoxide and hydrogen peroxide via the Haber-Weiss reaction. In order to determine whether this cleavage occurs in vivo, a chemiluminescence immunoblot system was developed to detect the presence of proteolysis products of transferrin or the related iron-binding protein, lactoferrin. Using this immunoblot system, we detected transferrin and lactoferrin cleavage products in bronchoalveolar lavage (BAL) samples from 21 of 22 and 20 of 21 cystic fibrosis (CF) patients, respectively. Three of eleven and two of nine BAL samples from individuals with other forms of chronic inflammatory lung disease had transferrin and lactoferrin cleavage products, respectively. Each patient in whom such products were detected was also infected with P. aeruginosa. No such products were detected in normal individuals. In the CF patients, there was no clear correlation between the extent of transferrin or lactoferrin cleavage and BAL neutrophil or P. aeruginosa concentration or the disease status of the patient. In contrast, in the non-CF patients with chronic inflammatory lung disease, transferrin and lactoferrin cleavage products were detected only in those BAL samples which contained the greatest concentration of both neutrophils and P. aeruginosa. These data provide evidence that P. aeruginosa- and/or human-derived protease cleavage of transferrin and lactoferrin occurs in vivo in the airways of individuals with CF and other forms of chronic lung disease, suggesting that this process could contribute to P. aeruginosa-associated lung injury in these patients. PMID- 8225582 TI - Legionella pneumophila growth restriction in permissive macrophages cocultured with nonpermissive lipopolysaccharide-activated macrophages. AB - Macrophages can be activated by lipopolysaccharides (LPS) from gram-negative bacteria to evince a number of biological activities, including increased resistance to intracellular infection by opportunistic bacteria. In the present study, intraperitoneal injection of LPS into A/J mice activated peritoneal macrophages so that they resisted subsequent in vitro infection with Legionella pneumophila. Coculture of these macrophages with those from nontreated A/J mice converted the entire population of cells from permissive to nonpermissive. This effect did not appear to be mediated by soluble factors released from the LPS treated macrophages, since the levels of interleukins-1 and -6 and tumor necrosis factor alpha produced by the macrophages were not found to be markedly elevated at the time when the macrophages from the LPS-treated mice were most effective in converting normal macrophages to nonpermissiveness. Furthermore, macrophages from mice injected intraperitoneally with either interferon or tumor necrosis factor alpha did not evince nonpermissiveness and also did not have the ability to convert normal spleen cells to nonpermissiveness. Polymyxin B, a known inactivator of LPS activity, did not inhibit the macrophages from the LPS-treated mice from inducing this resistance. It seemed unlikely that free LPS released from the macrophages mediated this effect. The results of this study thus showed that macrophages activated by LPS in vivo can evince nonpermissiveness for Legionella growth in vitro and also can induce macrophages from normal, permissive mice to become nonpermissive for Legionella growth in vitro. PMID- 8225583 TI - Induction of cytotoxic T lymphocytes against the Plasmodium falciparum circumsporozoite protein by immunization with soluble recombinant protein without adjuvant. AB - Immunization of mice with irradiated malaria sporozoites induces protection that is dependent on CD8+ T cells, and adoptive transfer of CD8+ cytotoxic T lymphocyte (CTL) clones against rodent malaria circumsporozoite (CS) protein and sporozoite surface protein 2 completely protects against sporozoite challenge. Thus, there are now efforts to develop vaccines that induce CTL against the CS protein and sporozoite surface protein 2. Until recently, it was thought that induction of CTL required production of target proteins within cells, breakdown of the proteins to peptides in the cytoplasm, and transport of the peptides to the cell surface in combination with class I major histocompatibility complex molecules. It has now been shown that immunization with peptides in Freund's complete adjuvant and with soluble protein in liposomes can induce CTL. To determine whether we could induce CTL against the Plasmodium falciparum CS protein by immunization with soluble protein, B10.BR mice were immunized intravenously, intraperitoneally, or intramuscularly with a recombinant P. falciparum CS protein called RLF mixed with the adjuvant DETOX (monophosphoryl lipid A, cell wall skeleton of Mycobacteria phlei, and squalane). Two weeks after the last dose, spleen cells from mice immunized intravenously, but not intraperitoneally or intramuscularly, had peptide-specific, major histocompatibility complex-restricted, CD8+ T-cell-dependent cytolytic activity against peptide 368-390 from the 7G8 P. falciparum CS protein. To determine whether the adjuvant was required for induction of the cytolytic activity, mice were immunized with RLF without adjuvant, and similar cytolytic activity was demonstrated. The finding that we could induce CTL by administration of soluble protein without adjuvant markedly broadens the possibilities for vaccinologists working to develop methods of inducing CTL in humans. PMID- 8225584 TI - Mobility of human neutrophils in response to Cryptococcus neoformans cells, culture filtrate antigen, and individual components of the antigen. AB - The mobility of human neutrophils (PMN) in response to encapsulated or nonencapsulated Cryptococcus neoformans cells or cryptococcal culture filtrate (CneF) and its components was studied by using a 48-well modified Boyden chamber. Encapsulated C. neoformans (isolate 184A) cells and CneF-184A stimulated directed migration of human PMN in the absence of serum (direct chemotactic activity) and activated a heat-labile component(s) in fresh human serum to become a chemoattractant(s) for human PMN (indirect chemotactic activity). At a 1:8 dilution (0.25 mg of carbohydrate per ml), CneF-184A displayed chemokinetic activity when assessed with a checkerboard assay. Nonencapsulated C. neoformans isolate 602 cells did not have direct chemotactic activity but did have indirect chemotactic activity. The capsule of C. neoformans is composed predominantly of glucuronoxylomannan (GXM). Purified GXM displayed both direct and indirect chemotactic activity. CneF-184A contains, in addition to GXM, a concanavalin A binding mannoprotein (MP), whereas CneF-602 contains no GXM but does contain MP. CneF-184A showed direct chemotactic activity and CneF-602 did not. Both CneF-184A and CneF-602 displayed indirect chemotactic activity for human PMN. In addition, purified MP from CneF-184A, like CneF-602, showed only indirect chemotactic activity. These results indicate that GXM contributes to the direct chemotactic activity of PMN observed with the whole encapsulated yeast cells and the unfractionated CneF derived from the encapsulated cells. Both MP and GXM from encapsulated C. neoformans cells mediate indirect chemotactic activity on human PMN. PMID- 8225585 TI - Cloning, expression, and mutagenesis of phosphatidylinositol-specific phospholipase C from Staphylococcus aureus: a potential staphylococcal virulence factor. AB - Staphylococcus aureus secretes a phosphatidylinositol (PI)-specific phospholipase C (PI-PLC) which is able to hydrolyze the membrane lipid PI and membrane protein anchors containing glycosyl-PI. The gene for PI-PLC (plc) was cloned from S. aureus into Escherichia coli. Oligonucleotide probes based on partial protein sequence and polyclonal antibodies raised against the purified protein were used to identify positive clones. E. coli transformed with a plasmid containing the plc gene expressed PI-PLC enzyme activity which was abolished by mutagenesis with a tetracycline resistance gene. The plc gene was present in all 15 S. aureus strains examined but not in any of 6 coagulase-negative staphylococcal species. The plc gene contained 984 bp and coded for a mature protein with a calculated molecular mass of 34,107 Da. Amino acid sequence comparisons indicated that the staphylococcal plc gene was similar (51 to 56%) to the PI-PLCs from Bacillus cereus, Bacillus thuringiensis, and Listeria monocytogenes. The recombinant PI PLC expressed in E. coli was purified and exhibited biochemical properties identical to those of the native PI-PLC from S. aureus. PI-PLC production was decreased in agr mutant strains of S. aureus. However, PI-PLC production by both agr+ and agr mutant strains exhibited a similar dependence on the type of medium used. These data suggested that PI-PLC production was regulated by both agr dependent and agr-independent mechanisms. PMID- 8225586 TI - Neutrophils are involved in acute, nonspecific resistance to Listeria monocytogenes in mice. AB - The importance of neutrophils in killing extracellular, pyogenic bacteria has long been established. However, there is only indirect evidence for a role for neutrophils in resistance against intracellular organisms. In this study, we directly demonstrate the involvement of neutrophils in defense against Listeria monocytogenes in normal C.B-17 immunocompetent and C.B-17 SCID mice. Because of the lack of sterilizing T-cell immunity, SCID mice are unable to completely eliminate listeriae systemically and become chronically infected. Both immunocompetent and SCID mice treated with a specific neutrophil-depleting monoclonal antibody during the early stages of Listeria infection were rendered remarkably sensitive to the organism, with a high level of mortality resulting from enhanced bacterial growth. At a late stage of infection in SCID mice, however, administration of neutrophil-depleting antibody did not affect mortality. In spite of the neutrophil depletion, other parameters of nonspecific immune function were normal. Macrophage infiltration to the site of infection and macrophage expression of major histocompatibility complex class II molecules were unaffected. Moreover, NK cell functions were normal as measured by infiltration to an infection site and gamma interferon production. These data demonstrate an important role for neutrophils in controlling the acute phase of Listeria infection, cooperating with, and yet independent of, macrophages and NK cells. PMID- 8225587 TI - Molecular characterization and expression of p23 (OspC) from a North American strain of Borrelia burgdorferi. AB - We have found that sera from patients with early stages of Lyme disease contain predominant immunoglobulin M reactivity to a major 23-kDa protein (p23) from Borrelia burgdorferi 2591 isolated in Connecticut. To characterize this immunodominant antigen, we cloned and sequenced p23 and found it to be 83% identical by nucleotide sequence and 75% identical by amino acid sequenced to pC (recently renamed OspC), an abundantly expressed protein on the outer surface of PKo, a European strain of B. burgdorferi (B. Wilske, V. Preac-Mursic, S. Jauris, A. Hofmann, I. Pradel, E. Soutschek, E.Schwab, G. Will, and G. Wanner, Infect. Immun. 61:2182-2191, 1993). In addition, immunoelectron microscopy localized p23 to the outer membrane, confirming that p23 is the strain 2591 homolog of OspC. The North American strain B31, commonly used in serologic assays for Lyme disease, does not express OspC. Northern (RNA) blot analysis detected low levels of ospC mRNA in B31, and DNA sequencing of the ospC gene from B31 revealed a 54 bp deletion in the upstream regulatory region, possibly accounting for the low transcriptional activity of ospC. The ospC coding region from B31 was cloned and antibody-reactive OspC was expressed in Escherichia coli. An immunoglobulin M enzyme-linked immunosorbent assay using recombinant OspC as the target antigen shows promise for the serodiagnosis of early stages of Lyme disease. PMID- 8225588 TI - Protective immunity is induced by a Borrelia burgdorferi mutant that lacks OspA and OspB. AB - A mutant of virulent Borrelia burgdorferi 297 was apparently selected for by long term storage at 5 degrees C. This mutant was found to lack the plasmid which encodes outer surface protein A (OspA) and OspB. In addition to the loss of the OspA and OspB proteins, the mutant lacked two lipoproteins, of 20 and 7.5 kDa, that were observed in the wild type. Since the mutant was not recovered from the tissues or blood of hamsters injected with the mutant, the mutant was determined to be noninfectious. Hamsters vaccinated with noninfectious mutant 297 were protected completely from challenge with virulent wild-type 297 spirochetes. Prechallenge sera from hamsters vaccinated with mutant 297 lacked antibodies to OspA and OspB, while those from hamsters vaccinated with virulent wild-type 297 or avirulent 297 exhibited antibodies to these proteins. Hamsters vaccinated with virulent wild-type 297 or mutant 297 elicited antibodies to OspC and a 39-kDa protein (P39), whereas hamsters vaccinated with avirulent 297 lacked these antibodies. These results suggest that OspC and/or P39 are important for the development of a protective immune response. Study of this mutant may elucidate factors important to the development of a Lyme disease vaccine. PMID- 8225589 TI - Differential susceptibilities of mice genomically deleted of CD4 and CD8 to infections with Trypanosoma cruzi or Trypanosoma brucei. AB - The role of CD4+ and CD8+ T cells in the surveillance of Trypanosoma cruzi or Trypanosoma brucei brucei was studied in mice which lacked CD4 or CD8 molecules and which were generated by embryonic stem cell technology. Whereas wild-type mice infected with T. cruzi (Tulahuen strain) displayed low levels of parasitemia and no mortality, striking increases in parasite growth and mortality occurred in both CD8- and CD4- mice. On the contrary, CD8- and, to a lesser degree, CD4- mice showed enhanced resistance to T. b. brucei. T-cell-dependent immunoglobulin G specific responses were produced in CD8- but not CD4- mice. Normal T-cell proliferative responses were measured in both CD4- and CD8- mice. Interleukin-4 production after concanavalin A or anti-CD3 monoclonal antibody stimulation was strikingly enhanced in CD8- but not CD4- spleen cells, whereas gamma interferon production was normal in both CD4- and CD8- spleen cells. Spleen and lymph node cells from CD8- (but not CD4-) mice at 20 days postinfection with T. cruzi had higher levels of interleukin-4 mRNA than the wild-type controls, as shown in a competitive polymerase chain reaction assay. On the other hand, CD4- (but not CD8 ) mice at 20 days postinfection with T. cruzi had lower levels of gamma interferon mRNA than the wild-type mice. PMID- 8225590 TI - Activation of the human complement alternative pathway by Listeria monocytogenes: evidence for direct binding and proteolysis of the C3 component on bacteria. AB - The capacity of the intracellular pathogen Listeria monocytogenes to activate the alternative pathway of human complement was examined. Incubation of L. monocytogenes with human serum in optimal conditions (20% Mg2+EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid]-chelated serum) consumed (31.3 +/- 3.9)% of C3 hemolytic activity and led to similar amounts of C3 deposition among the 27 strains tested, except for a rough mutant and the penicillin-induced L forms of strain EGD, which bound reduced amounts of C3. The same results were obtained with strains belonging to related species (L. innocua, L. seeligeri, L. welshimeri, and L. ivanovii). Direct evidence is provided that L. monocytogenes induces the deposition of C3b and its cleavage products iC3b and C3d through ester and amide linkages, as demonstrated by the analysis of the released products of radiolabelled purified C3 after treatment with hydroxylamine. Our results clearly demonstrate that L. monocytogenes activates the alternative pathway of human complement, suggesting that bacteria in the blood or in tissues of infected patients are opsonized and targeted to C3 receptor-bearing cells such as macrophages. PMID- 8225591 TI - Prevention of endotoxin-induced monokine release by human low- and high-density lipoproteins and by apolipoprotein A-I. AB - Interaction of endotoxin (lipopolysaccharide [LPS]) with human lipoproteins is known to prevent the LPS-induced activation of human monocytes and release of cytokines (monokines). LPS was exposed to lipoprotein classes separated by ultracentrifugation and to apolipoprotein A-I. Then monocytes were added, and the LPS activation of monocytes was determined by measuring the induced monokines. Failure of LPS to induce monokine release was called LPS inactivation caused by lipoproteins or apolipoproteins. The LPS inactivation is shown to be a function of low-density lipoproteins. High-density lipoproteins inactivate LPS to a much lesser extent. The very-low-density lipoproteins cannot inactivate LPS. Lipid components seemed not absolutely required for LPS inactivation, because purified human apolipoprotein A-I without its physiological lipid complement also inhibits LPS-induced monokine release. PMID- 8225592 TI - Characterization of Clostridium perfringens iota-toxin genes and expression in Escherichia coli. AB - The iota toxin which is produced by Clostridium perfringens type E, is a binary toxin consisting of two independent polypeptides: Ia, which is an ADP ribosyltransferase, and Ib, which is involved in the binding and internalization of the toxin into the cell. Two degenerate oligonucleotide probes deduced from partial amino acid sequence of each component of C. spiroforme toxin, which is closely related to the iota toxin, were used to clone three overlapping DNA fragments containing the iota-toxin genes from C. perfringens type E plasmid DNA. Two genes, in the same orientation, coding for Ia (387 amino acids) and Ib (875 amino acids) and separated by 243 noncoding nucleotides were identified. A predicted signal peptide was found for each component, and the secreted Ib displays two domains, the propeptide (172 amino acids) and the mature protein (664 amino acids). The Ia gene has been expressed in Escherichia coli and C. perfringens, under the control of its own promoter. The recombinant polypeptide obtained was recognized by Ia antibodies and ADP-ribosylated actin. The expression of the Ib gene was obtained in E. coli harboring a recombinant plasmid encompassing the putative promoter upstream of the Ia gene and the Ia and Ib genes. Two residues which have been found to be involved in the NAD+ binding site of diphtheria and pseudomonas toxins are conserved in the predicted Ia sequence (Glu-14 and Trp-19). The predicted amino acid Ib sequence shows 33.9% identity with and 54.4% similarity to the protective antigen of the anthrax toxin complex. In particular, the central region of Ib, which contains a predicted transmembrane segment (Leu-292 to Ser-308), presents 45% identity with the corresponding protective antigen sequence which is involved in the translocation of the toxin across the cell membrane. PMID- 8225593 TI - Evidence for asialo GM1 as a corneal glycolipid receptor for Pseudomonas aeruginosa adhesion. AB - Anti-gangliotetraosylceramide (anti-asialo GM1) and antiparagloboside monoclonal antibodies (MAbs) were used in immunofluorescence, immunoelectron-microscopic, and in vitro binding inhibition assays to determine whether either of the glycolipids was detectable in the normal cornea, whether levels changed following corneal scarification and either trypsin treatment or incubation in vitro with Pseudomonas aeruginosa, and whether either of the MAbs could competitively inhibit P. aeruginosa binding to cornea. No immunostaining above background for either glycolipid was observed in frozen, unfixed sections or in lightly fixed, K4M-embedded antibody-gold-labeled thin sections of normal cornea. In frozen sections of organ-cultured scarified cornea, no increased immunostaining for anti asialo GM1 or antiparagloboside reactivity was noted immediately or 60 min after corneal scarification. However, at 60 min after scarification and in vitro incubation of the eye with either trypsin or P. aeruginosa, enhanced immunostaining for both glycolipids was associated with cells within or immediately adjacent to the wound site. Trypsin increased immunoreactivity in the wound site more markedly compared with incubation with P. aeruginosa, but immunostaining was similarly localized with either treatment. No staining above background was seen in control sections. Similarly, with immunoelectron microscopy, increased immunogold-MAb staining for both glycolipids was seen on the plasma membranes of the wound-site cells of eyes incubated with either trypsin or P. aeruginosa compared with controls that were similarly immunostained but with the primary antibody either omitted or substituted with a nonspecific MAb. Competitive binding inhibition assays, in which the bacterial inoculum or the eye in organ culture was incubated with anti-asialo GM1 MAb prior to topical ocular application of the bacteria, showed significantly decreased P. aeruginosa adhesion compared with preparations similarly treated with phosphate-buffered saline or antiparagloboside MAb. These data provide evidence to support the hypothesis that asialo GM1, not paragloboside, serves as a receptor for P. aeruginosa binding to the scarified cornea of the adult mouse and spatially localizes both glycolipids in the wound site. PMID- 8225594 TI - Parasite virulence factors during falciparum malaria: rosetting, cytoadherence, and modulation of cytoadherence by cytokines. AB - To determine virulence factors of isolates of Plasmodium falciparum and the potential role of cytokines in cerebral malaria, 46 Malagasy patients presenting with cerebral (n = 10), severe (n = 10), and uncomplicated (n = 26) malaria were enrolled in a study. The capacity of 21 of 46 P. falciparum isolates to form rosettes in vitro and to adhere to human umbilical vein endothelial cells (HUVECs) that express intercellular adhesion molecule-1 receptors and to C32 amelanotic melanoma cells that express mainly CD36 receptors was investigated together with the effects of tumor necrosis factor alpha (TNF-alpha), granulocyte macrophage-colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and IL-6 alone and in two-by-two combinations on the cytoadherence of infected erythrocytes to HUVECs. Plasma levels of these cytokines were also measured in the patients at admission. The percentage of rosette formation was higher for the isolates from patients with cerebral (n = 6; 19.5%) and severe (n = 6; 30.5%) malaria than for those from patients with uncomplicated malaria (n = 9; 5%) (P < 0.002). The cytoadherence properties of the isolates did not differ among the three groups whatever the target cell used, but adherence to melanoma cells was systematically higher than that to HUVECs. Adhesion to HUVECs was increased more after TNF-alpha stimulation than after GM-CSF, IL-3, or IL-6 stimulation (P < 0.01). Only the combination of TNF-alpha and IL-3 enhanced cytoadherence more than TNF-alpha used alone (P < 0.02). No difference in the modulation of cytoadherence by cytokines was found in relation to the severity of the disease. TNF-alpha and IL-6 levels in peripheral blood were higher in the patients with cerebral and severe malaria than in the patients with uncomplicated malaria (P < 0.005). Most of the patients' sera contained little or no IL-3 or GM-CSF. Our results challenge the role of intercellular adhesion molecule-1 as the principal receptor mediating the cytoadherence of P. falciparum-infected erythrocytes and contrast with data obtained in the murine model. PMID- 8225595 TI - Prevention of renal scarring from pyelonephritis in nonhuman primates by vaccination with a synthetic Escherichia coli serotype O8 oligosaccharide-protein conjugate. AB - Rhesus monkeys were vaccinated with a synthetic Escherichia coli serotype O8 oligosaccharide-protein conjugate. Using our experimental pyelonephritis monkey model, we tested whether such immunization was protective against the renal damage from inflammation following experimental infection with a P-fimbriated O antigenically homologous E. coli strain. The vaccination did not significantly alter the duration of bacteriuria or interfere with the infection. However, the vaccine was efficient in renal protection, as vaccinated animals showed significantly less intratubular infiltration of neutrophils (P < 0.02) and the degree of renal scarring was also significantly less in these animals (P > 0.005) than in the control animals. Total kidney involvement in the vaccinated animals was 16.9%, compared with 32.5% in the control animals (P = 0.07). PMID- 8225596 TI - Expression of monocyte chemoattractant protein 1 (MCP-1) in adult periodontal disease: increased monocyte chemotactic activity in crevicular fluids and induction of MCP-1 expression in gingival tissues. AB - The present study shows that monocyte chemotactic activity in crevicular fluids increases with severity of the disease and that a monocyte chemoattractant, monocyte chemoattractant protein 1 (MCP-1), is expressed as the predominant cytokine of gingival tissues and their fibroblasts treated with Porphyromonas (Bacteroides) gingivalis lipopolysaccharide (P-LPS). High monocyte chemotactic activity in the crevicular fluids was neutralized significantly by antiserum specific for the JE/MCP-1 protein. Marked expression of the MCP-1 gene was observed in the gingival tissues of all adult periodontal patients tested, but not in those of healthy subjects. Monocyte chemotactic activity was observed in culture supernatants of human normal gingival tissues treated with P-LPS, and the chemotactic activity increased in a dose-related manner. Expression of MCP-1 in P LPS-treated human gingival fibroblasts was further examined. P-LPS induced the MCP-1 gene expression in a dose- and treatment time-dependent manner. The MCP-1 gene product in the culture supernatant was detected as two forms with molecular masses of 11,000 and 15,000 Da by immunoprecipitation with the specific antiserum. The MCP-1 gene expression was induced in the fibroblasts treated with interleukin-1 beta and tumor necrosis factor alpha, but not with interleukin-6. These results suggest that gingival fibroblasts can participate in monocyte recruitment in gingival tissues of adult periodontal patients via the MCP-1 gene product and that MCP-1 plays an important role in the inflammatory reaction in the disease. PMID- 8225597 TI - Chronic staphylococcal osteomyelitis: a new experimental rat model. AB - A rat model of chronic staphylococcal osteomyelitis was developed. Fibrin glue (5 microliters) and Staphylococcus aureus (2 x 10(6) CFU/5 microliters) were inoculated into the proximal metaphysis of the tibia. The rats were killed at intervals of between 1 and 6 months, and the tibias were removed. Induced lesions were evaluated by radiographic, macroscopic, and histological examinations and bacterial counts. Roentgenograms revealed osteomyelitis in more than 90% of the tibias. Gross bone pathology revealed skeletal deformation, new bone formation, abscesses, and draining skin fistulas in more than 80% of cases. Histological examination revealed osteomyelitis in more than 90% of cases, and bacterial counts were positive in 86% of cases. Only fibrin glue (5 microliters) was inoculated into controls. Controls showed no osteomyelitic lesions, and counts were negative in seven of eight control tibias. The main feature of this model is the use of fibrin glue instead of the sclerosing agents and foreign bodies used in other models. The model reproduces lesions similar to those of human posttraumatic osteomyelitis and can be reliably used in pathophysiological and therapeutic studies. PMID- 8225598 TI - Expression of the Salmonella virulence plasmid gene spvB in cultured macrophages and nonphagocytic cells. AB - Certain serotypes of salmonellae carry virulence plasmids that greatly enhance the pathogenicity of these bacteria in experimentally infected mice. This phenotype is largely attributable to the 8-kb spv regulon. However, spv genes are not expressed while bacteria grow in vitro. We now show that spvB, which is required for virulence, is expressed rapidly after Salmonella dublin is ingested by cultured J774 and murine peritoneal macrophages and that expression is not affected by the alkalinization of intracellular vesicles. The level of induction of spvB is reduced when macrophages are pretreated with gamma interferon. spvB is also expressed in human and canine epithelial cell lines and a human hepatoma cell line. In all cases, spvB expression is dependent on the spvR gene, just as it is in stationary-phase cultures in vitro. These data suggest that spv virulence genes are expressed by intracellular salmonellae in vivo in response to a signal that is common to the intracellular compartments of cells that are invaded by salmonellae. PMID- 8225599 TI - Relationship between Ileal symbiont intracellularis and porcine proliferative enteritis. AB - The relationship between Ileal symbiont (IS) intracellularis, formerly known as a Campylobacter-like organism, and porcine proliferative enteritis (PE) was studied by use of pigs with experimentally transmitted PE. Twenty one pigs were experimentally inoculated with homogenized ileal mucosa from a pig that died with PE, and 7 were maintained as uninoculated controls. Fecal samples were collected, and pigs were necropsied weekly postinoculation. Light microscopy and electron microscopy were used to examine tissues for lesions of PE and infectious agents. DNA was extracted from the fecal samples and assayed for the presence of sequences specific for IS intracellularis by dot blot hybridization and polymerase chain reaction amplification. IS intracellularis was detected by the polymerase chain reaction in the feces of 20 of 21 inoculated pigs but not in the feces of uninoculated pigs. Seven inoculated pigs but no uninoculated pigs were detected shedding IS intracellularis by dot blot hybridization. Shedding was detected 1 to 5 weeks after inoculation, and clinical signs were seen in the second to fifth weeks after inoculation. Few pigs without lesions of PE were found to shed IS intracellularis. There was a highly significant association between the presence of IS intracellularis in feces or tissue and the presence of microscopic proliferative lesions and between the severity of the lesions of PE and the percentage of IS intracellularis-infected intestinal crypts. Pigs that ceased shedding IS intracellularis were significantly less likely to have proliferative lesions. These and previous reports are consistent with the hypothesis that IS intracellularis is a necessary causative agent of PE. PMID- 8225600 TI - Cytokine induction by extracellular products of oral viridans group streptococci. AB - During an etiological study of Kawasaki disease (mucocutaneous lymph node syndrome [MCLS]), we found that dominant viridans streptococcal strains on tooth surfaces and in the throat of both MCLS patients and non-MCLS control children formed erythrogenic and biologically active, extracellular products. In this study, we demonstrated that erythrogenic culture supernatant concentrates of representative strains (two Streptococcus mitis and two Streptococcus oralis), when injected intravenously, induced serum tumor necrosis factor alpha, interleukin-6 (IL-6), and gamma interferon in muramyldipeptide- or Propionibacterium acnes-primed C3H/HeN mice. The concentrates also induced tumor necrosis factor alpha, IL-6, and thymocyte-activating factor (essentially IL-1) in murine peritoneal macrophage, human monocyte, and human whole-blood cultures. An erythrogenic, heat-labile extracellular protein fraction (F-1) that was concentrated from the culture supernatants of a representative S. mitis strain exhibited the above-mentioned cytokine-inducing activity. This partially purified F-1 fraction also induced thymocyte-activating factor and IL-6 in human umbilical vascular endothelial cell and gingival fibroblast cultures. PMID- 8225601 TI - Analysis of the roles of antilipopolysaccharide and anti-cholera toxin immunoglobulin A (IgA) antibodies in protection against Vibrio cholerae and cholera toxin by use of monoclonal IgA antibodies in vivo. AB - Secretory immunoglobulin A (IgA) antibodies (sIgA) directed against cholera toxin (CT) and surface components of Vibrio cholerae are associated with protection against cholera, but the relative importance of specific sIgAs in protection is unknown. A monoclonal IgA directed against the V. cholerae lipopolysaccharide (LPS), secreted into the intestines of neonatal mice bearing hybridoma tumors, was previously shown to provide protection against a lethal oral dose of 10(7) V. cholerae cells. We show here that a single oral dose of 5 to 50 micrograms of the monoclonal anti-LPS IgA, given within 2 h before V. cholerae challenge, protected neonatal mice against challenge. In contrast, an oral dose of 80 micrograms of monoclonal IgA directed against CT B subunit (CTB) failed to protect against V. cholerae challenge. A total of 80 micrograms of monoclonal anti-CTB IgA given orally protected neonatal mice from a lethal (5-micrograms) oral dose of CT. Secretion of the same anti-CTB IgA antibodies into the intestines of mice bearing IgA hybridoma backpack tumors, however, failed to protect against lethal oral doses of either CT (5 micrograms) or V. cholerae (10(7) cells). Furthermore, monoclonal anti-CTB IgA, either delivered orally or secreted onto mucosal surfaces in mice bearing hybridoma tumors, did not significantly enhance protection over that provided by oral anti-LPS IgA alone. These results demonstrate that anti-LPS sIgA is much more effective than anti-CT IgA in prevention of V. cholerae-induced diarrheal disease. PMID- 8225602 TI - Immunomodulation of human peripheral blood mononuclear cell functions by defined lipid fractions of Mycobacterium avium. AB - Mycobacterial fractions, some of which are associated with the cell envelope of Mycobacterium avium serovar 4, were assessed for their ability to affect various immunological functions of human peripheral blood mononuclear cells (PBM). Treatment of PBM with a total lipid fraction derived from M. avium serovar 4 resulted in a significant suppression of lymphoproliferative responsiveness to phytohemagglutinin stimulation at concentrations not affecting cell viability. Although a similar suppression was not observed when PBM were treated with purified serovar 4-specific glycopeptidolipids (GPL), treatment with the beta lipid fragment derived from the GPL did result in a significant suppression of phytohemagglutinin responsiveness. Further studies revealed that the total lipid fraction and the beta-lipid fragment were effective at significantly reducing the ability of human macrophages to restrict the intracellular growth of mycobacteria and at stimulating PBM to secrete prostaglandin E2. These same effects were not observed when purified GPL or the reduced oligosaccharide fragment of the GPL was used. Other studies revealed that the total lipid and purified GPL fractions were effective at stimulating tumor necrosis factor alpha release from human PBM, whereas the beta-lipid fragment was not. These results indicate that mycobacterial lipids have various immunomodulatory capabilities, depending upon their chemical nature and ability to interact with certain host cells. PMID- 8225603 TI - Human T cells recognize mycobacterial heat shock proteins in the context of multiple HLA-DR molecules: studies with healthy subjects vaccinated with Mycobacterium bovis BCG and Mycobacterium leprae. AB - Heat shock proteins (HSP) are considered to be important targets of the immune response to mycobacteria and, as such, relevant to subunit vaccine design. If HSP are major antigens in cell-mediated immunity, they should be recognized in the context of most of the HLA-DR molecules required for presentation of mycobacterial antigens to T cells. We tested peripheral blood mononuclear cells (PBMC) and T-cell lines from Mycobacterium leprae- and M. bovis BCG-vaccinated subjects for proliferation in response to the 18- and 65-kDa HSP of M. leprae, the 65-kDa HSP of M. bovis BCG, and the 70-kDa HSP of M. tuberculosis. Irrespective of HLA types, PBMC showing a strong response to M. leprae proliferated in response to mycobacterial HSP. HLA restriction analysis with T cell lines showed that the M. leprae 18-kDa HSP was recognized in the context of HLA-DR4, HLA-Dw4, and HLA-DR1 molecules. The T-cell lines recognized the M. leprae 65-kDa HSP in the context of all of the HLA-DR molecules expressed by autologous antigen-presenting cells, i.e., HLA-DR1, HLA-DR2, HLA-DR5, HLA-DR7, and importantly HLA-DR4 (HLA-Dw4 and HLA-Dw14), which is relevant to autoimmunity. The M. tuberculosis 70-kDa antigen was also presented to the T-cell lines by HLA-DR1, HLA-DR2, HLA-DR5, and HLA-DR7 molecules. In addition, this HSP was recognized in the context of the HLA-DRw53 molecule, which is frequently expressed in many regions where leprosy is endemic. The T-cell lines proliferating in response to a given HSP lysed autologous monocytes-macrophages pulsed with that HSP. The results demonstrate that PBMC from individuals immunized with M. leprae respond to mycobacterial HSP and that these HSP are presented to T cells by multiple HLA-DR molecules, a prerequisite for their application in the next generation of subunit vaccines. PMID- 8225604 TI - Two different types of ADP-ribosyltransferase C3 from Clostridium botulinum type D lysogenized organisms. AB - We examined production of ADP-ribosyltransferase C3 in 11 strains of Clostridium botulinum type C and D and their nontoxigenic derivatives. Antisera to C3 proteins of type C organisms divided C3 proteins roughly into at least two groups, bearing no relation to their bacterial types. The C3 gene of type D strain South African was isolated from a toxigenic phage library, and the complete sequence of the C3 gene was determined. The C3 protein of type D strain South African had 98% homology to the C3 protein of type C strain 003-9 and 66% homology to that of type D strain 1873. These results indicate that there are two types of C3 protein in type D organisms, as there are in type C organisms. PMID- 8225605 TI - Purification of Helicobacter pylori superoxide dismutase and cloning and sequencing of the gene. AB - The superoxide dismutase (SOD) of Helicobacter pylori, a pathogenic bacterium which colonizes the gastric mucosa, evoking a marked inflammatory response, was purified and characterized, and the N-terminal amino acid sequence was determined. The enzyme consists of two identical subunits each with an apparent molecular weight of 24,000. Analysis of the primary structure and inhibition studies revealed that H. pylori possesses a typical procaryotic iron-containing enzyme. No other isoenzymes could be detected. Indirect gold immunostaining of H. pylori SOD with a polyclonal antibody directed against the iron-containing SOD of Escherichia coli showed a surface-associated localization of the enzyme. The H. pylori SOD gene was cloned by functional complementation of a SOD-deficient E. coli mutant. Sequencing and alignment revealed striking homology to the following facultative intracellular human pathogens: Listeria ivanovii, Listeria monocytogenes, Coxiella burnetti, Porphyromonas gingivalis, Legionella pneumophila, and Entamoeba histolytica. An open reading frame of 642 bp encoding 214 amino acids was determined. There was no leader sequence detectable. Cloning of the H. pylori SOD gene is one of the prerequisites to investigation of its pathophysiological role in the defense against antimicrobial mechanisms of polymorphonuclear granulocytes. PMID- 8225606 TI - Toxicity of staphylococcal enterotoxins potentiated by lipopolysaccharide: major histocompatibility complex class II molecule dependency and cytokine release. AB - The biological effects of staphylococcal enterotoxins (SE), potentiated by bacterial lipopolysaccharide (LPS), were studied with mice. Control animals survived the maximum dose of either SE or LPS, while mice receiving both agents died. SEA was 43-fold more potent than SEB and 20-fold more potent than SEC1. The mechanism of toxicity was further examined with transgenic mice deficient in major histocompatibility complex class I or II expression. Class II-deficient mice were resistant to SEA or SEB. However, class I-deficient animals were less susceptible to SEA (30% lethality) than wild-type mice (93% lethality). In vitro stimulation of T cells from the three mouse phenotypes by SEA correlated well with toxicity. T cells from transgenic or wild-type mice were similarly responsive to SEA when presented by irradiated, wild-type mononuclear cells. These data confirmed that the toxicity of SE was mainly exerted through a mechanism dependent on the expression of major histocompatibility complex class II molecules. Toxicity was also linked to stimulated cytokine release. Levels in serum of tumor necrosis factor alpha, interleukin-6, and gamma interferon peaked 2 to 4 h after the potentiating dose of LPS but returned to normal within 10 h. Concentrations of interleukin-1 alpha were also maximal after 2 h but remained above the background for up to 22 h. Relative to the levels in mice given only SEA or LPS, the levels in serum of tumor necrosis factor alpha, interleukin-6, and gamma interferon increased 5-, 10-, and 15-fold, respectively, after injections of SEA plus LPS. There was only an additive effect of SEA and LPS on interleukin-1 alpha concentrations. PMID- 8225607 TI - Isolation of Brucella abortus ssb and uvrA genes from a genomic library by use of lymphocytes as probes. AB - Brucella abortus proteins from virulent S2308 expressed from a pBluescript II SK- genomic library stimulated peripheral blood mononuclear (PBM) cell proliferation from cattle vaccinated with B. abortus S19. The method described here permits a rapid and directed approach to isolate genes encoding antigens of B. abortus that interact with lymphocytes primed to the living bacterium. The supernatants from the bacterial host JM109 (DE3) were cultured with freshly isolated bovine PBM cells. A total of 300 clones were evaluated. Ten clones were identified that stimulated T-lymphocyte proliferation. Among them, one clone with a 2.5-kb insert stimulated T-lymphocyte proliferation in all three animals, suggesting that the proteins encoded by genes within this fragment may represent immunodominant antigens. DNA sequencing of this clone reveals two large open reading frames (ORFs). ORF II has a high degree of similarity to the Escherichia coli ssb gene, which codes for the single-stranded DNA binding protein. ORF I, in the opposite direction to ORF II, shows similarity to the N terminus of the E. coli uvrA gene, which codes for one of the three subunits of the E. coli ABC excision nuclease. The observation that the PBM cells recognized and proliferated in response to proteins expressed from single clones provides a novel strategy to select bacterial antigens that may prove useful in designing alternative vaccines against brucellosis. PMID- 8225608 TI - Comparison of naturally acquired and vaccine-induced antibodies to Haemophilus influenzae type b capsular polysaccharide. AB - The objective of this study was to assess qualitative differences in the types of Haemophilus influenzae type B (Hib) capsular polysaccharide (polyribosylribitol phosphate [PRP]) antibodies induced in children 15 to 27 months of age by (i) natural exposure, (ii) PRP vaccine, and by (iii) PRP-diphtheria toxoid conjugate vaccine, (iv) PRP-group B Neisseria meningitidis outer membrane vesicle conjugate vaccine, and (v) Haemophilus type B oligosaccharide conjugate vaccine (HbOC). The highest levels of total Hib-PRP antibody measured by radioimmunoassay and immunoglobulin G (IgG) measured by enzyme-linked immunosorbent assay were seen after HbOC immunization. IgG1 Hib-PRP antibodies predominated in all groups, and there were no differences between the groups in the proportion of IgG and IgA Hib PRP antibodies. However, the proportions of IgM differed significantly by group. The highest proportions of IgM occurred in naturally acquired antibody and after PRP vaccine, and the lowest proportion occurred after HbOC vaccine. IgG light chain V kappa type alpha PRP antibody was present in all groups, and the level correlated with the total IgG Hib-PRP antibody level. Therefore, HbOC induced the highest concentrations of V kappa II type alpha PRP antibody, and the naturally acquired antibody group had the lowest levels. IgG light-chain V kappa III antibody levels were also highest in the HbOC group, but there was no correlation between V kappa III antibody levels and total amount of IgG Hib-PRP antibody. These data demonstrate qualitative differences in the antibody repertoires induced by natural exposure, the Hib-PRP vaccine, and each of the different Hib conjugate vaccines. We doubt that there are major differences in the protection afforded by these different antibody repertoires, because these differences do not appear to correlate with differences in protective efficacy in older children. PMID- 8225609 TI - Role of the physical state of Salmonella lipopolysaccharide in expression of biological and endotoxic properties. AB - Lipopolysaccharide (LPS) extracted from three strains of Salmonella typhimurium, i.e., the rough Re mutant SL1102, the rough Ra mutant TV119, and the smooth strain SH4809, was first electrodialyzed (eLPS) and then divalent cation deprived by EDTA treatment and finally made monomeric by deoxycholate solubilization. The removal of excess detergent by extensive dialysis in the absence of mineral cations resulted in the reassociation of LPS subunits into monodisperse micelles of reduced aggregation number (dLPS) as estimated by electron microscopy and gel filtration chromatography. For all LPS chemotypes tested, the developed procedure reproducibly results in stable and clear solutions of dLPS in concentrations of up to 100 mg/ml. The dLPS and eLPS preparations possessed the same reactivity with monoclonal antibodies (MAbs) raised against different LPS domains. The 100% lethal dose in galactosamine-sensitized mice of 0.01 microgram for the smooth eLPS was from 10- to 100-fold lower than that of dLPS at 0.1 to 1.0 microgram. dLPS from both the smooth strain and the Ra mutant had a significantly reduced capacity to activate the proenzyme cascade in the Limulus amoebocyte lysate assay in comparison with the slightly reduced activity of dLPS from the Re mutant. In contrast, dLPS as well as the deoxycholate-dispersed and then diluted eLPS from the smooth strain had a higher mitogenic activity on splenocytes than eLPS. The results indicate that the biological and endotoxic properties of LPS are significantly influenced by the physical state of its aggregates in aqueous solutions. The approach developed for production of a stable and dispersed form of LPS should further assist in investigation of LPS properties and interpretation of the data of endotoxic research. PMID- 8225610 TI - Identification of Legionella pneumophila genes required for growth within and killing of human macrophages. AB - Legionella pneumophila was mutagenized with Tn903dIIlacZ, and a collection of mutants was screened for defects in macrophage killing (Mak-). Of 4,564 independently derived mutants, 55 (1.2%) showed a reduced or complete lack in the ability to kill HL-60-derived human macrophages. Forty-nine of the Mak- mutants could be assigned to one of 16 DNA hybridization groups. Only one group (9 of the 10 members) could be complemented for macrophage killing by a DNA fragment containing icm and dot, two recently described L. pneumophila loci that are required for macrophage killing. Phenotypic analysis showed that none of the mutants were any more sensitive than the wild type to human serum, oxidants, iron chelators, or lipophilic reagents nor did they require additional nutrients for growth. The only obvious difference between the Mak-mutants and wild-type L. pneumophila was that almost all of the Mak- mutants were resistant to NaCl. The effects of LiCl paralleled the effects of NaCl but were less pronounced. Resistance to salt and the inability to kill human macrophages are linked since both phenotypes appeared when Tn903dIIlacZ mutations from two Mak- strains were transferred to wild-type backgrounds. However, salt sensitivity is not a requisite for killing macrophages since a group of Mak- mutants containing a plasmid that restored macrophage killing remained resistant to NaCl. Mak- mutants from groups I through IX associated with HL-60 cells similarly to wild-type L. pneumophila. However, like the intracellular-multiplication-defective (icm) mutant 25D, the Mak- mutants were unable to multiply within macrophages. Thus, the ability of L. pneumophila to kill macrophages seems to be determined by many genetic loci, almost all of which are associated with sensitivity to NaCl. PMID- 8225611 TI - Salmonella typhimurium delta aroA delta aroD mutants expressing a foreign recombinant protein induce specific major histocompatibility complex class I restricted cytotoxic T lymphocytes in mice. AB - Recombinant Salmonella typhimurium aroA aroD mutants which expressed ovalbumin were constructed. The two expression constructs used were based on either pUC18 or pBR322. The pBR322-based construct was more stable in vitro and in vivo than the pUC-based construct. Salmonellae containing the stable pBR322-based plasmid induced major histocompatibility complex (MHC) class I-restricted cytotoxic T lymphocytes (CTL), in contrast to salmonellae containing the pUC18-based expression construct. The priming of MHC class I-restricted CTL was increased by multiple immunizations. The study described in this report suggest that S. typhimurium delta aro mutants have the capacity to induce MHC class I-restricted CTL against carried antigens and that MHC class I-restricted CTL responses require stable in vivo expression of the target antigen. Further, the results indicate that the Salmonella typhi delta aro mutants currently undergoing evaluation in studies with humans may be good carriers of viral antigens with CTL determinants. PMID- 8225612 TI - Regulation of inflammation-primed activation of macrophages by two serum factors, vitamin D3-binding protein and albumin. AB - A very small amount (0.0005 to 0.001%) of an ammonium sulfate [50% saturated (NH4)2SO4]-precipitable protein fraction of alpha 2-globulin efficiently supported inflammation-primed activation of macrophages. This fraction contains vitamin D3-binding protein essential for macrophage activation. Comparative macrophage activation studies with fetal calf serum, alpha 2-globulin fraction, 50% (NH4)2SO4 precipitate, and purified bovine vitamin D3-binding protein revealed that fetal calf serum and alpha 2-globulin fraction appear to contain an inhibitor for macrophage activation while ammonium sulfate precipitate contains no inhibitor. This inhibitor was found to be serum albumin. When bovine serum albumin (25 micrograms/ml) was added to a medium supplemented with 0.0005 to 0.05% (NH4)2SO4 precipitate or 1 to 10 ng of vitamin D3-binding protein per ml, activation of macrophages was inhibited. PMID- 8225613 TI - ADP-ribosylation of rainbow trout (Oncorhynchus mykiss) actin by botulinum C2 toxin. AB - Intracellular actin of rainbow trout macrophages was ADP-ribosylated by botulinum C2 toxin, which is composed of two nonlinked protein components, component I and trypsinized component II. The actin in the supernatants of various tissue homogenates of the trout was also directly ADP-ribosylated by component I of C2 toxin, indicating that fish actin other than those of land vertebrates is susceptible to enzymatic modification by component I of C2 toxin. PMID- 8225614 TI - Antigenic switching of TSA 417, a trophozoite variable surface protein, following completion of the life cycle of Giardia lamblia. AB - Expression of TSA 417, the predominant cysteine-rich variable surface protein of Giardia lamblia WB clone C6 trophozoites, did not change during encystation in vitro. However, in vitro excystation of cysts derived in vitro or in vivo consistently produced TSA 417 nonexpressing trophozoite populations, suggesting that completion of the life cycle leads to antigenic switching. PMID- 8225615 TI - Hemin utilization is related to virulence of Streptococcus pneumoniae. AB - Streptococcus pneumoniae is a causative agent for bacterial pneumonia, otitis media, meningitis, and bacteremia. Mechanisms for acquisition of iron by this organism under low-iron conditions were investigated. Siderophore production was not detected by either chemical or biological methods. Its utilization of iron containing compounds found in human hosts was tested. Both hemin and hemoglobin supported the full growth of S. pneumoniae in a culture lacking other iron sources, while lactoferrin and transferrin failed to do so. A mutant defective in hemin utilization was isolated and was less virulent than wild-type S. pneumoniae in experimental animals. PMID- 8225616 TI - Circulating tumor necrosis factor alpha (TNF), soluble TNF receptors, and interleukin-6 in human subacute bacterial endocarditis. AB - Cell surface components of viridans streptococci and enterococci have been shown to stimulate the release of tumor necrosis factor alpha (TNF) and interleukin-6 from monocytes/macrophages. In the sera from 10 patients with subacute enterococcal or streptococcal endocarditis, however, the levels of both cytokines were low or undetectable, with elevated TNF levels on admission in 3 patients with complicated disease. Soluble TNF receptor levels were significantly elevated compared with those of healthy controls. When patients with malaria were used as a control group of acute intravascular infection with high circulating TNF values, the ratio between soluble TNF receptors and TNF on admission was significantly greater in the patients with subacute bacterial endocarditis. Besides different amounts of circulating TNF, enhanced TNF receptor shedding may have an important role in the pathogenesis of subacute versus acute clinical disease following human intravascular infection. PMID- 8225617 TI - Comparative immunochemistry of two fragments from domains Ib and III of Pseudomonas aeruginosa exotoxin A. AB - Two rabbit polyclonal antisera have been produced by immunization with two fragments corresponding to sequences 392 to 404 and 392 to 613 of Pseudomonas aeruginosa exotoxin A. Both antisera inhibit the ADP-ribosyltransferase activity of exotoxin A but do not inhibit its NAD-glycohydrolase activity. In addition, only the second antiserum was capable of neutralizing exotoxin A cytotoxicity in cell culture and in vivo. Consequently, the common sequence 392 to 404 of the two fragments is not a neutralizing epitope and such an epitope should reside within residues 405 to 613 of exotoxin A. The sequence 392 to 404 was shown to be hidden in the native molecule, and the results suggest that this sequence is most likely in close proximity to residues involved in eukaryotic elongation factor 2 binding. PMID- 8225618 TI - Mutations in the promoter spacer region and early transcribed region increase expression of staphylococcal enterotoxin A. AB - The mechanism leading to increased production of staphylococcal enterotoxin type A (SEA) in mutant Staphylococcus aureus FRI722 compared within its wild-type parent strain, FRI100, was examined. Sequence analysis revealed two mutations in the upstream promoter region of FRI722 at nucleotides -28 and +3 with respect to the transcriptional initiation site at An sea translational fusion of the upstream region of FRI722 to the structural gene from FRI100 showed an increase in sea expression by Northern (RNA) analysis and in SEA production by Western (immunoblot) analysis. To independently evaluate the effect of each mutation, site-directed mutagenesis was done and revealed that each mutation was responsible for an increase in SEA production. PMID- 8225619 TI - Virulence of two Streptococcus pyogenes strains (types M1 and M3) associated with toxic-shock-like syndrome depends on an intact mry-like gene. AB - The major virulence factor of Streptococcus pyogenes, the M protein, is positively regulated at the transcriptional level by mry in the M type 6 strain studied. We show here that in two S. pyogenes strains isolated from cases of toxic-shock-like syndrome, a type M1 strain and a type M3 strain, an mry-like gene is also required for resistance to phagocytosis. PMID- 8225620 TI - Passive immunization against cytomegalovirus in allograft recipients. The Rotterdam Heart Transplant Program experience. AB - We analyzed the results of passive immunization against CMV in 146 heart transplant recipients. The 65 seronegative recipients were prophylactically treated with anti-CMV immunoglobulins during and after the operation. Twenty-nine of these 65 patients received a seropositive donor heart. CMV infection occurred in 21/65 seronegative and in 40/81 seropositive recipients (difference not significant). The incidence of CMV infection in seronegative recipients of a CMV matched donor heart (3/34) was significantly lower than in seronegative recipients of a positive donor heart and lower than in seropositive recipients, but no significant difference in infection rate was found between the two latter groups (18/29 vs. 40/81). Although primary infection more frequently resulted in CMV disease than secondary infection (11/21 vs. 10/40) no difference in incidence of disease was noted between seronegative and seropositive patients (11/65 vs. 10/81), nor was there a difference in the severity of symptoms following primary or secondary infection. There was a higher incidence of CMV disease in all patients who received a heart from a seropositive donor versus a seronegative donor. However, after transplantation of a heart from a seropositive donor the incidence (27%) of CMV disease observed in our passively immunized seronegative patients was the same as in the patients with naturally acquired seropositivity. There was no difference in the prevalence of coronary artery disease between patients with and without CMV infection or disease. We conclude that using the current passive immunization scheme the occurrence of CMV infection and disease is largely dependent on the serostatus of the donor. PMID- 8225621 TI - Cephadroxil promotes vaginal colonization with Escherichia coli. AB - The aim of this study was to examine whether a vaginal Escherichia coli colonization, mimicking the one seen in UTI-prone females, could be induced by local cephadroxil administration. When five adult cynomolgus monkeys were given a vaginal flush with a P-fimbriated E. coli strain, none became persistently colonized. When such colonization attempts were preceded by cephadroxil administration a persistent colonization occurred in 9/10 experiments. Cephadroxil also promoted a spread of fecal E. coli strains to the vagina. Reduction of the anaerobic vaginal flora can explain the breakdown of the colonization resistance. Clinical observations suggest that accumulation of E. coli around the urethral orifice increases the risk of UTI. Therefore antibiotics which promote such colonization may increase the risk for UTI in susceptible patients. From this point of view antibiotics such as cephadroxil may be less suitable for treatment of UTI-susceptible patients. PMID- 8225622 TI - Variability of serum concentrations of trimethoprim and sulfamethoxazole during high dose therapy. AB - Serum kinetics of trimethoprim and sulfamethoxazole were studied in 23 patients during oral and i.v. treatment of Pneumocystis carinii pneumonia. Daily doses of 15-22 mg/kg trimethoprim and 75-110 mg/kg sulfamethoxazole were given every 6 h. Despite administration of a loading dose of twice the regular dose, serum trough concentrations continuously rose from 12 h to 96 h by 63% for trimethoprim and 102% for sulfamethoxazole. After 4-6 days mean trough concentrations of trimethoprim and sulfamethoxazole were 7.7 +/- 3.0 and 198 +/- 74 mg/l, with individual values of < 4.6 and < 103 mg/l in two patients and > 11.4 and > 307 mg/l in two others. Patients treated orally or i.v. had similar serum levels. However, large interindividual variability was observed despite weight-specific dosing. Administration of a loading dose did not prevent accumulation of serum levels of trimethoprim and sulfamethoxazole over several days of treatment. PMID- 8225623 TI - Serum concentrations of interferon gamma, interleukin-6 and neopterin in patients with infectious mononucleosis and other Epstein-Barr virus-related lymphoproliferative diseases. AB - Serum levels of interferon gamma, interleukin-6 and neopterin were determined in 15 patients with different forms of Epstein-Barr virus-associated diseases: acute self-limiting infectious mononucleosis, chronic active infectious mononucleosis and X-linked lymphoproliferative syndrome. In patients with acute type of infection, neopterin, interferon gamma and interleukin-6 were elevated in nearly all patients. In contrast, the situation was less clear-cut in the other EBV associated diseases; particularly interleukin-6 was undetectable in most cases. The results suggest that concomitant measurement of these diverse immune activation markers may provide interesting insights into the interactions between the virus and the host, and may also lead to therapeutic consequences. PMID- 8225624 TI - Antimicrobial prophylaxis in hysterectomies: a double-blind, randomized, comparative study of a single dose of piperacillin (2 g) versus a single dose of amoxicillin-clavulanic acid (2.2 g). AB - A double-blind, randomized comparative study of piperacillin (2 g) versus amoxicillin-clavulanic acid (2.2 g) as a single dose 30 minutes before the initiation of hysterectomy was performed. A total of 595 patients (of which 307 were in the piperacillin group) were evaluable for efficacy. Infectious complications were infrequent in both arms. One case of (mild) sepsis was observed in the piperacillin group and two cases of wound infection were observed in the amoxicillin-clavulanic acid group. Urinary tract infection was observed in 5.5% of the patients in the piperacillin group and in 2.4% of the amoxicillin clavulanic acid group. A relatively high incidence of asymptomatic bacteriuria was seen in both groups: 11.8% in the piperacillin group and 8.7% with amoxicillin-clavulanic acid. A marked difference was seen between the two different hospital locations: a 15% incidence in the hospital where midstream urine was used for culture (Sittard), versus 5% in the hospital where catheter urine was used (Geleen). It is concluded that both antibiotics are associated with a low rate of infectious complications and that catheter urine must be used for sample collection. PMID- 8225625 TI - Ceftazidime versus ceftazidime plus tobramycin in febrile neutropenic children. AB - Although the effectiveness of antibiotic monotherapy in febrile neutropenic patients remains unproven, ceftazidime has been shown previously to be effective monotherapy for the empiric treatment of selective patients. The efficacy and safety of ceftazidime versus ceftazidime plus tobramycin was evaluated in the treatment of febrile children (range 8 months to 18 years) with neutropenia secondary to cancer chemotherapeutic agents. Of the evaluable 89 patients, 45 received ceftazidime and 44 received ceftazidime plus tobramycin for 5 to 10 days. At the end of therapy, 30 (67%) of the 45 ceftazidime-treated patients were clinically cured compared with 38 (86%) of 44 combination-treated patients. Thirteen (29%) of the patients treated with ceftazidime failed to respond clinically to treatment, versus four (9%) of the patients treated with ceftazidime/tobramycin (p = 0.046). This study suggests that ceftazidime as monotherapy in febrile neutropenic children may be inferior to combination therapy for optimal clinical response in these patients. PMID- 8225626 TI - Penetration of ceftazidime into human pancreas. AB - The use of antibiotics in patients with severe acute pancreatitis (stage II and III) is indicated since bacterial complications are the most common cause of death in these patients. In the present study the penetration of ceftazidime into pancreatic juice, into healthy and chronically inflamed pancreatic tissue as well as into necrotic regions in cases of severe acute pancreatitis was investigated. A peak concentration of 12.9 +/- 5.9 mg/l was found 60 min after intravenous administration of 35 mg/kg of the drug, which is 32% of the corresponding serum levels. Pancreatic tissue concentrations varied between 9 and 79% of the corresponding serum levels, depending on the stage of inflammation. After five days of antibiotic treatment with doses of 2 g t.i.d., concentrations between 1.8 and 6.9 mg/kg were detected even in pancreatic necroses. This suggests that sufficient antibacterial levels of ceftazidime were present in all pancreatic compartments analyzed following administration of common therapeutic dosages. Therefore, from a pharmacokinetic point of view, ceftazidime could be a potentially effective drug for the treatment of pancreatitis. PMID- 8225627 TI - Nosocomial outbreak of legionellosis in a rehabilitation center. Demonstration of potable water as a source. AB - Ten patients from a rehabilitation center were admitted to hospital with serious respiratory infections within ten weeks. An outbreak of Legionnaire's disease was suspected based on the epidemic and atypical manifestation of pneumonia and could be proven microbiologically. Pulmonary and extrapulmonary complications included respiratory failure, lung abscess, transitory renal impairment in five patients and acute renal failure requiring dialysis in one, tetraparesis caused by peripheral neuropathy and acute psychosis. Three patients died despite immediate institution of therapy with erythromycin. Legionella pneumophila serogroup 1 subtype Pontiac was isolated from a bronchial lavage sample of one patient and from the water supply of the rehabilitation center. Monoclonal antibody subtyping and restriction endonuclease analysis were performed on both environmental and patient isolates. Potable water was identified as the source of the outbreak based on identical patterns on restriction endonuclease analysis. Despite thermic and chemical disinfection with chlorination (up to 15 ppm) in the rehabilitation clinic, an eleventh case of Legionnaire's disease was detected 11 months later. PMID- 8225628 TI - Anaerobic bacteremia: a retrospective four-year analysis in general medicine and cancer patients. AB - Anaerobic bacteremia was studied in 32 medical patients (mean age 72 years) in a four-year retrospective analysis. Malignancy was the most common underlying disease and probable portal of entry. The gastrointestinal tract was affected most often, followed by the respiratory and urinary tracts. Bacteremia occurred either following invasive (non surgical) procedures or spontaneously. The clinical course ranged from asymptomatic bacteremia, to mild febrile illness, to sepsis and septic shock (two, 12, 16 and two patients, respectively). The case fatality rate was 25%. The causative organisms were Clostridium and Bacteroides species. All organisms isolated were susceptible to chloramphenicol. Early diagnosis and prompt treatment may reduce mortality in cases of anaerobic sepsis. PMID- 8225629 TI - The effect of clindamycin gel insert in periodontal pockets, as observed on smears and cultures. AB - This study is aimed at the evaluation of a 1% clindamycin hydrochloride containing gel on the microbial flora of periodontal pockets deeper than 5 mm. In order to achieve that purpose, 20 patients with pocketing in the premolar-molar regions were selected. Active and placebo gel were inserted once during the first 2 weeks of this experimental study. Microbial samplings were performed 1, 2, 4 and 12 weeks after the experiment started. The samples were submitted to microscopic examination and also to culture. Changes in the microbial content of the periodontal pockets treated by subgingival scaling and clindamycin 1% gel were significant, compared with those obtained with subgingival scaling and placebo gel, particularly with respect to anaerobic black-pigmented bacteria and the motile gram-negative flora. However, after 3 months, most of the treated cases were recolonized by the same initial species, though never at pre clindamycin levels. In the light of this study, it will be concluded that the use of a small amount of clindamycin hydrochloride inserted into a periodontal pocket, once a week for 2 weeks as a complement to periodontal subgingival scaling, is beneficial in the treatment of adult periodontitis, by eliminating more effectively the microbial pocket colonization. PMID- 8225630 TI - Postoperative cauda syndrome caused by Staphylococcus aureus. AB - Toxic shock syndrome (TSS) is a well-defined clinical syndrome attributed to certain exotoxins produced by Staphylococcus aureus. The acute episode is often characterized by a toxic encephalopathy, possibly caused by direct neurotoxicity of these exotoxins, although this mechanism has never been proven. We describe a patient who developed TSS, meningitis and cauda equina syndrome simultaneously several days after lumbar laminectomy. A space-occupying lesion was excluded. Enterotoxin C-producing S. aureus was cultured from the surgical wound and the cerebrospinal fluid (CSF). The patient recovered from TSS but remained partially paralyzed. Presumably the cauda equina syndrome was caused by neurotoxic effects of the intrathecally produced S. aureus exotoxins. This case provides evidence for the neurotoxic effects of TSS-associated S. aureus exotoxins. PMID- 8225631 TI - Cerebral Salmonella typhimurium abscess in a patient with a stroke. AB - A 59-year-old woman with renal failure was admitted in a coma with seizures. Computerized tomography (CT) revealed a frontal mass in the right cerebral hemisphere and on day 12 new low density areas had appeared in both the right and left hemisphere. Blood cultures on day 13 grew Salmonella typhimurium, and the patient died the next day. An autopsy showed cerebral infarction with abscess formation. The elusive features of non-typhoid Salmonella infections make it advisable to obtain blood and faecal cultures from susceptible patients with fever or focal disorders of unknown aetiology. PMID- 8225632 TI - Fatal Pasteurella multocida infection in a systemic lupus erythematosus patient. PMID- 8225633 TI - Ideal age for measles vaccination with persisting maternal antibody, human immunodeficiency virus infection and protein-calorie malnutrition. PMID- 8225634 TI - Antimicrobial activity of josamycin against erythromycin-resistant staphylococci as compared to roxythromycin and clarithromycin. AB - In an in vitro study 246 clinical isolates of erythromycin-resistant staphylococci from six hospitals in Austria were investigated for susceptibility to josamycin and other, newer macrolide antibiotics, e.g. roxithromycin and clarithromycin. 71 strains of Staphylococcus aureus showed an MIC > or = 4 mg/l and 100 strains of S. aureus showed an MIC > or = 256 mg/l. In addition, 25 strains of coagulase-negative staphylococci resistant to erythromycin at an MIC of > or = 4 mg/l were investigated. At an MIC of 2 mg/l 57% of the erythromycin resistant strains of S. aureus were inhibited by josamycin, 25% by clarithromycin and 11.6% by roxithromycin. At an MIC of 2 mg/l 13.3% of erythromycin-resistant coagulase-negative staphylococci were inhibited by josamycin, 10.7% by clarithromycin and 9.3% by roxithromycin. This study suggests that josamycin is still active in vitro against more than 50% of erythromycin-resistant strains of S. aureus. This drug is also more active than roxithromycin and clarithromycin against erythromycin-resistant S. aureus. PMID- 8225635 TI - Biliary tract infections treated with ciprofloxacin. AB - Thirty-two patients with severe biliary tract infections (cholecystitis and cholangitis) were treated with ciprofloxacin intravenously followed by oral ciprofloxacin. Complete clinical and bacteriological cure has been observed in 28 out of 32 patients and therapy failure occurred in four patients. Overall, no major adverse effects were encountered. These data suggest that intravenous ciprofloxacin followed by oral administration is an effective and safe agent for the therapy of severe biliary tract infections. PMID- 8225636 TI - Clarithromycin 250 mg b.i.d. for 5 or 10 days in the treatment of adult patients with purulent bronchitis. AB - This Phase III, double-blind, randomized, multicenter study compared the safety and efficacy of clarithromycin, 250 mg b.i.d. administered for either 5 or 10 days, in the treatment of 221 adult patients with purulent bronchitis. Clinical evaluations were performed pre-treatment, during treatment, post-treatment, and at a follow-up visit. Both regimens were well tolerated and effective. No significant differences were observed between patients treated for 5 days and those treated for 10 days in post-treatment, clinical success (98%, 83/85; 99%, 73/74, respectively), or bacterial eradication (100%, 37/37; 94%, 33/35) rates, or in the number of patients reporting drug-related adverse events (16%, 18/112; 24%, 26/109, respectively). Clarithromycin administered for 5 days was at least as safe and as effective as a 10-day regimen in the treatment of purulent bronchitis. PMID- 8225637 TI - A comparative study of clarithromycin and amoxycillin suspensions in the treatment of pediatric patients with acute otitis media. AB - This phase III, single-blind, randomized, multicentre clinical trial compared the safety and efficacy of clarithromycin and amoxycillin in the treatment of otitis media in pediatric patients. Two hundred and fifty-nine patients aged 1-12 were prescribed suspensions of clarithromycin (132 patients) or amoxycillin (127 patients). Both suspensions were prescribed at a dose of 125 mg for children weighing less that 25 kg or at 250 mg for children weighing more than 25 kg, but three doses of amoxycillin per day were given, while only two doses clarithromycin per day were required. Each drug was administered for approximately 5 days. Clinical evaluations were performed pre-treatment (Study Day 1), at the end of treatment (Study Days 6-9), and post-treatment (Study Days 28-32). At the end of treatment, 91 out of 114 evaluable patients (80%) had clinical cures with clarithromycin, while 71 out of 105 evaluable patients (68%) had clinical cures with amoxycillin (p = 0.057). Clinical success rates were 96% for both treatments (110/114, clarithromycin; 101/105 amoxycillin). Adverse events related to the study medications occurred in four of 132 patients receiving clarithromycin (3%) and eight out of 127 subjects receiving amoxycillin (6%). Three patients discontinued treatment due to adverse events, all three receiving amoxycillin. At the doses administered, clarithromycin given twice daily was as safe and effective as given three-times-daily in the treatment of acute otitis media in pediatric patients. PMID- 8225638 TI - Fluid transport and bacterial penetration along root canal fillings. AB - Convective transport of water from the coronal to the apical end of obturated root canals was determined by the movement of an air bubble in a capillary glass tube connected to the apex of the experimental root section using a headspace pressure of 120 kPa (1.2 atm). Water transport through existing voids in the obturated canals could be measured reproducibly in this way. The root canals of 60 human maxillary canines were filled with gutta-percha and sealer by the cold lateral condensation technique. Thirty of these were first exposed to a small motile bacterium, Pseudomonas aeruginosa, growing in a reservoir at the coronal end of each root. After 50 days, two specimens allowed penetration of bacteria to a reservoir at the apical end. All the roots were then assessed quantitatively for convective transport of water. The results were divided into three defined categories: 39 obturated canals were in the 'bacteria tight' category, 14 canals in the 'slight leakage' and 7 canals in the 'gross leakage' category. The two specimens that showed bacterial penetration fell into the slight and gross leakage categories. The previous test for bacterial passage did not statistically influence the fluid transport pattern of these roots which was measured subsequently. These findings indicate that fluids transport through obturated root canals, most of which do not allow the passage of bacteria. PMID- 8225639 TI - The effect of using files with altered tips in a sonic handpiece: an in vitro study. AB - The objective of this study was to establish the effect of altering the non cutting tips on Shaper files during canal preparation with the MM1500 Sonic Air handpiece. A total of 90 simulated canals in clear resin blocks of various angles and positions of curvature were prepared using Shaper files activated by an MM1500 Sonic Air handpiece. Thirty canals were prepared with unmodified files having non-cutting tips, 30 canals were prepared with files which had their tips removed completely and a further 30 canals were prepared with files which had tips reduced by half. All canals were prepared using an identical preparation procedure which involved a linear filing motion and an anticurvature technique in curved canals. The parameters investigated included loss of canal length, weight loss from the blocks, smoothness of canal walls, transportation and overall shape of the prepared canals. The effect of altering the file tips was to increase significantly the loss of working distance and the prevalence of canal aberrations such as zips and danger zones. Altering the tips also had a detrimental effect on the pattern and extent of transportation as well as widening canals excessively in their apical third. Overall, under the conditions of this study, alterations to the non-cutting tips of Shaper files was counterproductive and resulted in canals displaying many adverse features. Conversely, retention of the non-cutting tips allowed canal preparation to proceed rapidly and effectively producing canal shapes which displayed many desirable features. PMID- 8225640 TI - Light-cured glass ionomer cement as a retrograde root seal. AB - A light-cured ionomer cement was investigated as a retrograde root seal, without a retrograde cavity. This was compared with the material used in a retrograde cavity, and with a conventional glass ionomer cement, as a seal. The adaptation and sealing ability of the test materials were assessed using a confocal optical microscope with a fluorescent dye. The root canals of 40 extracted human single rooted teeth were prepared and filled with gutta-percha and sealer. All the teeth were subsequently apicected, then divided into four equal groups. In one group, a retrograde cavity was prepared, and the light-cured glass ionomer cement was placed as a retrograde root filling. No retrograde cavities were prepared in the three remaining groups. The light-cured glass ionomer cement was applied directly onto the apicected root face. Two different thicknesses of light-cured glass ionomer cement were tested, a thin layer (approximately 1 mm) in one group, and a thicker layer (> 1 mm) in another group. A conventional glass ionomer cement was used in the last group, and applied directly onto the root face in a single thickness (approximately 1 mm). In the group where the light-cured glass ionomer cement was used in a retrograde cavity, the material was often well adapted to one cavity wall, but gaps were found on the opposite wall. The light-cured and conventional glass ionomer cement retrograde root seals were well adapted to the root face, regardless of the thickness of material used. The thinly applied (approximately 1 mm) light-cured glass ionomer cement retrograde root seals permitted the least leakage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225641 TI - Radicular cyst affecting a root-filled human tooth: a long-term post-treatment follow-up. AB - Apical periodontitis is caused primarily by microorganisms residing in the root canals of affected teeth. Nevertheless, there is convincing evidence implicating other independent factors that adversely affect the outcome of conventional root canal therapy. In this paper, morphological evidence is presented in support of the potential role of two endogenous factors that may interfere with post endodontic healing of the periapex. The specimens consisted of a surgical biopsy of an asymptomatic periapical lesion which persisted for a follow-up period of 44 months. The biopsy was processed for correlated light and electron microscopy. The lesion was characterized by the presence of a large central lumen lined by a stratified squamous epithelium. The most striking feature of the lesion was the presence of vast numbers of cholesterol crystals which congregated in the connective tissue surrounding the cyst cavity. Extensive light and electron microscopic investigation of the apical part of the root canal and the lesion failed to reveal the presence of microorganisms. These findings strongly suggest that intrinsic factors like the accumulation of certain tissue break-down products such as cholesterol crystals, and the cystic condition of the lesion itself, can adversely affect the healing process of the periapex following root canal therapy. Consequently, such apical lesions can remain refractory to conventional endodontic therapy for long periods of time. PMID- 8225642 TI - Neural changes in ulcerative and hyperplastic pulpitis: a transmission electron microscopic study. AB - The purpose of this study was to examine at ultrastructural level the neural changes in human apical pulps of teeth clinically diagnosed as having ulcerative or hyperplastic pulpitis. Seventeen specimens were prepared from six teeth with ulcerative or hyperplastic pulpitis and four specimens from two sound teeth. After routine procedures the specimens were examined under an electron microscope for changes in the structure of myelinated and unmyelinated nerve fibres. Although the teeth used in this study responded positively to vitality test, the results revealed that moderate to severe degenerative changes occurred in the nerve fibres of the apical pulps of the teeth showing these two diseases. PMID- 8225643 TI - A comparative sealability study of different retrofilling materials. AB - The purpose of this study was to compare the sealing ability of high-copper amalgam, glass ionomer cement, and silver glass ionomer cement as retrofilling materials when used with and without varnish. Seventy extracted human single rooted anterior teeth were instrumented and the root canals obturated with laterally condensed gutta-percha and sealer. The apical 3 mm of each root was resected and a 2 mm-deep apical Class I preparation cut. The roots were then randomly allocated to six groups of 10 teeth and retrofilled with one of the experiment materials. Group I comprised high-copper amalgam; group 2, high-copper amalgam with cavity varnish; group 3, glass ionomer cement without varnish; group 4, glass ionomer cement with varnish; group 5, silver glass ionomer cement without varnish; and group 6, silver glass ionomer cement with varnish. Two groups of five roots each were used as positive and negative controls. The root surfaces were coated with nail polish and specimens immersed in 2% methylene blue solution for 14 days. After vertical sectioning, dye penetration was examined under a stereomicroscope. The results of the study were analysed statistically. The sealing ability of the conventional glass ionomer cement with varnish was significantly better while dye penetration was higher in silver glass ionomer cement without varnish group than the other groups (P < 0.01). PMID- 8225644 TI - Maxillary permanent incisors with two root canals: a report of two cases. AB - The root canal treatment of permanent maxillary incisors with two root canals is reported. Two cases are presented, one success and one failure, with discussion regarding the different outcome, highlighting the need for careful radiographic investigation and patient management. PMID- 8225645 TI - Dental anxiety: a cause for possible misdiagnosis of tooth vitality. AB - Fear, anxiety and anticipation of pain are prepotent emotional and cognitive mediators of pain behaviour. As pain is often used as a parameter for evaluation of tooth vitality, existing dental anxiety may occasionally lead to possible misdiagnosis. A representative case is presented in which dental anxiety led to an initial misdiagnosis of endodontic pathology. PMID- 8225646 TI - Bladder cancer among dialysis patients with schistosomiasis. PMID- 8225647 TI - Adequacy of continuous ambulatory peritoneal dialysis. AB - Adequacy of continuous ambulatory peritoneal dialysis (CAPD) and its variants is assessed by clinical outcomes, biochemical parameters and clearance parameters. Clinical outcomes lack specificity and probably sensitivity. Nevertheless, they constitute the "gold standard" to which any other method assessing dialysis adequacy must be compared. Biochemical parameters are both non-sensitive and non specific and cannot be used to assess dialysis adequacy. Clearance of small molecular weight azotemic substances (urea, creatinine) presents considerable computational problems and interpretative difficulties. In preliminary studies, clearance studies have been able to differentiate between peritoneal dialysis patients having symptoms of inadequate dialysis and those clinically adequately dialyzed. Among population outcomes (morbidity, maintenance of peritoneal dialysis for long periods, hospitalization rate, mortality), only mortality seems to be associated with low clearances in retrospective studies. Prospective multicenter studies comparing clearance values to clinical outcomes are needed to evaluate clearance studies as methods of assessing peritoneal dialysis adequacy. PMID- 8225648 TI - Technical and clinical evaluation of a new asymmetric polysulfone membrane (Biosulfane). AB - First generation asymmetric polysulfone membranes had high hydraulic permeability (kf = 40 ml/h/mmHg/sqm) but a low diffusive permeability due to the hydrophobic nature and wall thickness of 75-100 microns. We have tested a new polysulfone membrane with a wall thickness of 40 microns in a series of in vitro and in vivo dialysis session experiments. The new "Biosulfane" membrane presented a Kf of 45.8 with constant performance up to 240 mins. The koA was 760 and the clearance value at 350 ml/min of Qb in hemodiafiltration was 255 ml/min for urea, 210 for creatinine, 225 for phosphate, 76 for inulin. In high flux dialysis the clearances were similar except for inulin which was 32% lower due to the lower convection amount. Beta-2 microglobulin clearance was 22 ml/min in high flux dialysis and 37 in hemodiafiltration. Solute sieving coefficients were close to 1 for the majority of the studied solutes in a wide range of molecular weights and slight variations were observed for charged solutes due to Donnan's effect. The sieving for Inulin was 0.96 while that for Beta-2 microglobulin was not measurable due to a large molecule adsorption on the inner structure of the fibres. The good performances of this membrane are probably due to reduced wall thickness and a consequent improvement in diffusive permeability to small size solutes. PMID- 8225649 TI - A model for non-invasive estimation of in vivo dialyzer performances and patient's conductivity during hemodialysis. AB - On-line monitoring of hemodialysis sessions requires a non-invasive estimation of the parameters concerning the patient's status and the dialyzer performances. We describe here a model based on a new method for non-invasive dialysance and patient conductivity measurements. In this technique the same probe measures alternately the conductivity at the dialysate inlet and outlet for two different dialysate conductivity values. From these data, an appropriate model allows to determine the patient's conductivity as well as the effective dialysance of ionised solutes, that is to say the dialysance corrected for recirculation. A strong correlation is evidenced between the effective dialysance measured by this method and the urea clearance measured by conventional methods (r = 0.98 for in vitro solutions; r = 0.82 in vivo situations). PMID- 8225650 TI - Continuous veno-venous hemofiltration in critically ill patients with multiple organ failure. AB - 18 critically ill patients, with multiple organ failure (MOF) (from shock either septic, n = 15, or cardiogenic, n = 3), oliguria and increase in BUN and creatinine were treated with pump driven, high flux continuous veno-venous hemofiltration (CVVH). Replacement fluids were administered in predilution mode. All patients were under respiratory support and vasoactive drugs, and received early nutritional support (N input: 0.2-0.3 g/kg/day). Mean duration of treatment was 9.2 days and mean ultrafiltrate production was 21.4 l/day; treatment resulted in a significant reduction of both urea nitrogen and creatinine blood levels (-20 and -40% of initial values respectively) in spite of a very severe catabolism. The total amount of urea nitrogen removed through CVVH ranged from 15 to 73 g/day (mean 33.5), the median value of urea nitrogen clearance was 12.8 ml/min with a median ultrafiltration coefficient of 0.8. The mean duration of hemofilters was 69 hours (38-108); the efficacy of filters remained stable throughout the entire lifespan and changes were made in case of sudden decrease of ultrafiltration (< ml/min). No major complication was observed in over than 4000 hours of treatment. Pump driven, high flux CVVH proved effective in the control of water electrolyte balance and metabolic homeostasis in a group of critically ill, hemodynamically unstable, catabolic patients with MOF and acute renal failure. In no case we had to add intermittent hemodialysis or to use hemodiafiltration. The constant extracorporeal blood flow and the stable efficacy of hemofilters allowed an easy control of the overall effectiveness of this technique. PMID- 8225651 TI - Effect of intradialytic parenteral nutrition on quality of life in hemodialysis patients. AB - The purpose of this study was to determine the effect of intradialytic parenteral nutrition (IDPN) on the quality of life of malnourished patients with end-stage renal disease on high-efficiency and high-flux hemodialysis therapy. Patients, who met the Medicare eligibility requirements for IDPN, were asked to fill out and extensive questionnaire covering several measures of quality of life prior to initiating and again after completing four months of IDPN therapy. Although the IDPN improved serum albumin levels significantly, aside from improved sleep patterns, no significant improvements in quality of life could be demonstrated. The mortality rate of these enrolled patients was as high as 28% within 4 months. We concluded that the limited amount of nutrition delivered over the course of a short dialysis session may not be enough to appreciably change the lives of our malnourished patients. PMID- 8225652 TI - Side effects of hybrid liver support therapy: TNF-alpha liberation in pigs, associated with extracorporeal bioreactors. AB - During acute liver failure, hybrid liver support therapy could serve as a bridge to liver transplantation. In this desired temporary use, immune competent cell responses, such as the production of cytokines, might be of limiting relevance. We have investigated the Tumor Necrosis Factor-alpha (TNF) liberation in two models using pigs, connected with an extracorporeal bioreactor with homologous hepatocytes: TNF liberation was measured in arterial plasma during a 4 day perfusion time in untreated animals, model (i), and during short term perfusion of hepatectomized pigs in model (ii). Animals four days after catheter implantation in model (i) had TNF values of < 5 pg/ml. After connecting the system without hepatocytes, TNF rose to 9.7 +/- 2 within 120 min and rose further to 32.6 +/- 6 pg/ml within 4 hours after filling the system with the homologous hepatocytes. After 24 hours of continuous perfusion and during four days of perfusion, the TNF levels were lowered to baseline levels. In model (ii), TNF rose to 220 +/- 130 pg/ml within 180 min and decreased to 110 +/- 10 pg/ml within six hours, whereas controls without hepatocytes showed mean levels with a maximum of 120 +/- 20 pg/ml. In both models, there was no correlation between TNF levels and clinical abnormalities such as fever or shock symptoms. There is evidence for an activation of blood cells during experimental extracorporeal hybrid support. No typical side effects were, however, observed. Thus, TNF mediated extracorporeal cell activation does not appear to limit the application of homologous hybrid liver support therapy. PMID- 8225653 TI - Growth factors and biological supports for endothelial cell lining: in vitro study. AB - Endothelial cell covering over the vascular prosthesis luminal surface is a process that may require the presence of growth factors (GFs) and extracellular matrix supports. Endothelialization could be improved by combining both GFs and an extracellular matrix analog. In the present study, different biological substrates made of type I or IV collagens, gelatin, fibronectin, fibrin, laminin, chondroitin sulphate, heparan sulphate, heparin or hyaluronic acid were used to support endothelial cell culture. An endothelial cell growth supplement (ECGS) was incorporated in (group 1) or overlaid on (group 2) the substrates; or present in medium (group 3); or absent (group 4). GF binding assay using 125I bFGF showed that more GF remained combined to the substrates in group 2 than those in group 1. Growth and morphology of human umbilical vein endothelial cells were sequentially analyzed in vitro for 8 days using DNA (nuclei counts) and F-actin labelings. Growth was relatively stable for the first 48 hours, later in groups 1, 2 and 4, cell death was observed on all the substrates except for fibronectin. Growth failure could be related to the degradation or inefficient release of ECGS. In group 3, growth increased and confluency was reached within 5-8 days on all the substrates except for gelatin and type I collagen. Confluent cells containing actin filaments were organized on glycoproteins and disorganized on glycosaminoglycans and fibrin. Despite that glycoproteins can enhance cell adhesion and lining pattern, GFs continually delivered in a fresh soluble form seem to be the appropriate condition to obtain an endothelial cell lining. PMID- 8225654 TI - Selective inhibition by phorbol 12,13-dibutyrate of the alpha 1-receptor-mediated positive inotropic effect. AB - Influence of the protein kinase C activator phorbol 12,13-dibutyrate on the alpha 1- and beta-adrenoceptor-mediated positive inotropic effect was studied in the rabbit ventricular myocardium. Phorbol 12,13-dibutyrate (10(-8)-10(-6) M) inhibited the positive inotropic effect mediated by alpha 1-adrenoceptors in a concentration-dependent manner, while the positive inotropy mediated by beta adrenoceptors was not affected by phorbol 12,13-dibutyrate up to 3 x 10(-7) M. Phorbol 12,13-dibutyrate at 10(-6) M decreased the beta-mediated effect, but the extent of inhibition was less than that of alpha 1-mediated effect produced by 10(-8) M phorbol 12,13-dibutyrate. Thus, the inhibition induced by phorbol 12,13 dibutyrate was 100-fold more selective for alpha 1- than for beta-mediated inotropy. Phorbol 12,13-dibutyrate at 10(-7) M increased the basal force of contraction in some preparations, but decreased it at 3 x 10(-7) M and higher in a concentration-dependent manner. In membrane fractions derived from the rabbit ventricular muscle, phorbol 12,13-dibutyrate did not affect the specific binding of [3H]prazosin. A nonhydrolyzable GTP analogue GTP gamma S shifted the epinephrine-induced displacement curve of [3H]prazosin to the right, but phorbol 12,13-dibutyrate did not affect the curve. Accumulation of [3H]inositol monophosphate induced by alpha 1 stimulation was inhibited by phorbol 12,13 dibutyrate. These findings indicate that phorbol 12,13-dibutyrate may induce the selective uncoupling of the myocardial alpha 1-receptor stimulation to activation of phospholipase C, and inhibit selectively the alpha 1-mediated positive inotropy. PMID- 8225655 TI - Plasma ANP and cyclic GMP after physical exercise in patients with mitral valve disease and in healthy subjects. AB - Plasma levels of both atrial natriuretic peptide (ANP) and cyclic GMP are elevated in patients with various heart diseases as compared to healthy subjects. In this study patients with advanced mitral valve disease (Group A) and healthy subjects (Group B) were exposed to symptom-limited upright stepwise physical exercise on a cycle ergometer. Concentrations of ANP and cyclic GMP were measured in plasma at rest (20 min in supine position) or 5 min after physical exercise by specific radioimmunoassays. Here we show that short dynamic exercise caused a significant increase in plasma levels of ANP and cyclic GMP, in both groups. In Group A strong correlation between plasma ANP and cyclic GMP was found at rest (r = 0.91, P < 0.001, n = 11) and after physical exercise (r = 0.85, P < 0.001, n = 11). In contrast, there was no correlation between plasma concentrations of ANP and cyclic GMP in Group B at rest (r = -0.16, P > 0.05, n = 10) or after exercise loading (r = 0.14, P > 0.05, n = 10). Absolute increases in circulating levels of both substances were not found to correlate in either group. These data suggest that exercise-induced elevations in plasma cyclic GMP may be due not only to ANP release but also to an as yet undetermined factor, possibly EDRF/NO. PMID- 8225656 TI - Effects of alpha-human atrial natriuretic peptide in guinea-pig isolated heart. AB - The aim of the present investigation has been to ascertain whether or not atrial natriuretic peptides (ANP) can exert a direct effect on myocardial contractility. Alpha-human ANP (alpha-hANP) concentrations ranging from 1 pM to 50 nM have been used to perfuse guinea-pig isolated hearts in a non-recirculating Langendorff apparatus. A dual concentration-related effect has been induced by alpha-hANP on myocardial function. A maximal increase of +LV dP/dtmax (+56%; P < 0.001) has been observed when guinea-pig hearts were perfused with 100 pM alpha-hANP, whereas a 25% decrease (P < 0.01) occurred with 50 nM alpha-hANP. Similar effects have also been induced by alpha-hANP on the coronary flow rate (CFR). A significant CFR increase (maximal at 10 pM alpha-hANP) was induced by picomolar concentrations of alpha-hANP, whereas a progressive decrease, which was maximal ( 28%; P < 0.01) at 50 nM alpha-hANP, was observed with nanomolar concentrations of the peptide. No effects have been observed on heart rate. These results suggest that ANP has direct effects on both vascular and myocardial muscle cells. Coronary vasoconstriction induced by nanomolar concentrations of ANP can contribute to the cardiodepression, whereas ANP in picomolar concentrations can induce a coronary vasodilation which is not coupled with the enhanced myocardial contractility. The latter is the likely expression of a direct effect of the peptide on myocardial function. PMID- 8225657 TI - The effects of oral ibopamine in patients with mild heart failure--a double blind placebo controlled comparison to furosemide. The Ibopamine Study Group. AB - The effect of ibopamine and furosemide in 130 patients with NYHA Class I and II heart failure were studied in a parallel, double-blind, randomized placebo controlled multi-centre trial. Ibopamine 200 mg b.i.d. was compared to furosemide 40 mg q.d. and placebo. A 1- to 2-week single-blind run-in period was followed by an 8-week double-blind treatment period. Reproducible treadmill exercise test times with the modified Naughton-Balke protocol were required for randomization. Exercise times increased significantly in comparison to the placebo group after 8 weeks of therapy for both the furosemide group (1.2 min, P < 0.035) and the ibopamine group (1.3 min, P < 0.025). Neither furosemide nor ibopamine affected quality of life assessments. Adverse clinical experiences were generally mild and similar in frequency amongst the three treatment groups. The results of this study show the usefulness of both ibopamine and furosemide as monotherapy in patients with mild congestive heart failure. PMID- 8225658 TI - Exercise-induced ventricular arrhythmias in patients with healed myocardial infarction. AB - BACKGROUND: Controversy exists about the clinical and prognostic significance of exercise-induced ventricular arrhythmias late after myocardial infarction. The aim of the study was to identify the main clinical and prognostic features of exercise-induced ventricular arrhythmias in out-patients with healed Q-wave myocardial infarction. METHODS: The study population was 777 consecutive patients who underwent a symptom-limited (Bruce protocol) treadmill test from May 1988 to January 1991 after myocardial infarction (at least 1 year). Clinical and exercise data were prospectively entered in a computerized database and retrospectively two different groups were selected: (1) 228 patients with exercise-induced ventricular arrhythmias; (2) 549 patients without. Incidence and morphology of exercise-induced ventricular arrhythmias, various exercise parameters and a follow-up were evaluated. RESULTS: Patients with exercise-induced ventricular arrhythmias were older (P < 0.001), had higher blood pressure (P < 0.03) and peak exercise rate pressure product (P < 0.00) than the others. No difference was found in the incidence of exercise-ischaemia: either symptomatic or not. When simple (< or = 2 Lown) versus complex (> or = 3 Lown) exercise-induced ventricular arrhythmias were considered, the latter were more frequent in patients with anterior myocardial infarction, shorter exercise duration (P < 0.001) and lower exercise rate pressure product, lower ejection fraction and lower incidence of exercise-induced ischaemia. In the follow-up (mean 24 +/- 13 month) there were 24 deaths: five (2.2%) in patients with exercise-induced ventricular arrhythmias and 19 (3.4%) in patients without. Cardiac event rate was similar in both groups. CONCLUSIONS: We conclude that in out-patients with healed myocardial infarction exercise-induced ventricular arrhythmias are quite frequent, but they are not associated with exercise-induced ischaemia, either symptomatic or not. Exercise-induced ventricular arrhythmias seem to be related to age or peak workload. Moreover patients with these arrhythmias have no adjunctive negative risk on prognosis. PMID- 8225659 TI - Intracardiac thrombosis diagnosed by echocardiography in childhood: predisposing and etiological factors. AB - Eleven cases of intracardiac thrombi caused by different factors including protein-C deficiency are presented for discussion of the etiology and predisposing factors of intracardiac thrombi during infancy and childhood, and to stress the importance of protein-C deficiency as an etiological factor. Thrombi were localised in the left heart in five patients and right heart in five patients. One patient had both-sided thrombi. Four of our patients had dilated cardiomyopathy, one had mitral valve hypoplasia, and one had pulmonary valvar stenosis as the predisposing factors for thrombus formation. In three patients whose cardiac anatomies were completely normal, we determined protein-C deficiency as an etiological factor of thrombus formation. One of these had congenital protein-C deficiency and the other two had acquired temporary protein C deficiency due to sepsis. In conclusion we recommend that protein-C deficiency should be investigated as an etiological factor in all cases of intracardiac thrombi irrespective of whether or not another predisposing factor is identified. PMID- 8225660 TI - What factors determine indium-111 antimyosin monoclonal antibody uptake in patients with myocardial infarction? AB - The intensity of indium-111 antimyosin monoclonal antibody uptake for visualization of myocardial infarction seems partially dependent on the state of the infarct related coronary artery. The aim of this study is to determine the factors which could account for the monoclonal antibody uptake variability. For this purpose, we investigated 27 patients (mean age 52.7 +/- 9.6 years) with a first proven myocardial infarction, by monoclonal antibody scintigraphy and coronary arteriography within the same period of time (7.12 +/- 6 days). The monoclonal antibody uptake was quantified by the heart/lung ratio on images recorded 24 h after injection. The infarct size was quantitatively estimated on wall motion analysis of twelve segments in 30 degree right anterior-oblique view with a radial method. The infarct related coronary artery state was assessed by the Thrombosis in Myocardial Infarction grade and the functional characteristics of collateral vessels by Rentrop's classification. These three variables as well as location of myocardial infarction, left ventricular ejection fraction, administration of a thrombolytic therapy, delay between myocardial infarction and monoclonal antibody scintigraphy were studied using non parametric test, or by linear regression method in order to determine whether these factors would influence the heart/lung ratio. None of these parameters except infarct size was related to heart/lung ratio. Consequently, monoclonal antibody uptake is only dependent on the extent of infarcted myocardium and the intensity of uptake cannot predict the patency of an infarct related coronary artery. PMID- 8225661 TI - Physiologic multivalvular regurgitation during pregnancy: a longitudinal Doppler echocardiographic study. AB - Valvular function, assessed by Doppler technique, has not been extensively investigated during normal pregnancy. To prospectively study this feature, 18 normal pregnant women were followed during their pregnancies and puerperium, with serial clinical and pulsed-continuous Doppler echocardiographic examinations. In four gestational periods and the puerperium, we analysed: (a) ventricular and atrial dimensions, as well as valve annular diameters; (b) prevalence and characteristics of trivial valvular regurgitations. During pregnancy, slight but significant increases of the four cardiac chamber dimensions and valve annular diameters were observed, except for the aortic ring. The prevalence of physiologic valvular regurgitation in early pregnancy (mitral, 0%; tricuspid, 38.9%; pulmonary, 22.2%; aortic, 0%), was similar to a control group of 18 healthy non-pregnant women. As pregnancy evolved, there was a progressive and significant increase of multivalvular regurgitation, maximal at full-term (mitral, 27.8%; tricuspid, 94.4%; pulmonary, 94.4%, P < 0.05 vs. early pregnancy). Aortic regurgitation was not detected in any stage of pregnancy. In the puerperium, mitral regurgitation resolved, but tricuspid and pulmonary regurgitation were still significantly prevalent (83.3% and 66.7%, respectively, P < 0.05 vs. early pregnancy). It is concluded that physiologic multivalvular regurgitation is frequent in pregnancy, mainly involving right-sided valves in late gestational periods, occasionally persisting in the early puerperium. Chamber enlargement, valve annular dilatation, and increased prevalence of trivial valve regurgitation are time-related events during normal pregnancy, resulting from a reversible cardiac remodeling process induced by physiologic volume overload. These aspects should be considered for a correct interpretation of Doppler echocardiographic findings in pregnant women with suspected heart disease. PMID- 8225662 TI - Endomyocardial biopsy in right ventricular cardiomyopathy. AB - Right ventricular cardiomyopathy is characterized by a progressive myocyte loss and fibro-fatty substitution of the right ventricle. The aim of our study was to assess the diagnostic accuracy of right ventricular endomyocardial biopsy. Using an imaging analyser system, histomorphometric parameters of myocytes, interstitium, fibrous tissue and fatty tissue were evaluated on endomyocardial biopsy from 30 patients with arrhythmogenic right ventricular cardiomyopathy, 29 patients with dilated cardiomyopathy and 30 control patients. The percent area of myocytes decreased from 78.10 +/- 7.34 in control to 63.39 +/- 9.22 in dilated cardiomyopathy (P < 0.05) and to 47.28 +/- 15.01 in arrhythmogenic right ventricular cardiomyopathy (P < 0.01). Fibrous tissue increased from 8.10 +/- 3.89 in control to 21.80 +/- 9.29 in dilated cardiomyopathy (P < 0.05) and to 24.60 +/- 11.37 in arrhythmogenic right ventricular cardiomyopathy (P < 0.05). Fatty tissue varied from 0.33 +/- 1.44 in control and 0.07 +/- 0.31 in dilated cardiomyopathy to 13.30 +/- 17.30 in arrhythmogenic right ventricular cardiomyopathy (P < 0.05). Fatty tissue was a feature of arrhythmogenic right ventricular cardiomyopathy (67% of patients vs. 6% of control and dilated cardiomyopathy patients). Diagnostic values typifying arrhythmogenic right ventricular cardiomyopathy, obtained by excluding any overlapping between confidence intervals in the three groups, were: myocytes < 44.95%; fibrous tissue > 40.38%, and fatty tissue > 3.21%, with 67% sensitivity and 91.53% specificity for at least one parameter. In conclusion, a significant difference between arrhythmogenic right ventricular cardiomyopathy, dilated cardiomyopathy and control exists in terms of amount of myocytes, fibrous tissue and fatty tissue. Presence of fatty tissue and fibrous tissue exceeding 3.21% and 40.38%, respectively should be considered highly suspect for arrhythmogenic right ventricular cardiomyopathy in right ventricular endomyocardial biopsy. PMID- 8225663 TI - Acute anuric renal failure: a complication of combined thrombolytic and antithrombotic therapy. AB - Thrombolytic agents are routinely given to patients with acute myocardial infarction, often in combination with heparin. Genitourinary hemorrhage is an uncommon adverse effect of systemic thrombolysis, and acute azotemic complications are distinctly rare. We describe a case of genitourinary hemorrhage after combined thrombolytic and antithrombotic therapy leading to acute bilateral ureteral obstruction and then briefly review renal complications of systemic thrombolysis. PMID- 8225664 TI - Syncope and long QT syndrome with an initially normal QT interval. AB - We present two patients who were admitted to our Hospital because of repetitive syncopal attacks. The study previous to their admission included electrocardiograms in which the QT interval was normal. During their admission, an striking lengthening of the QT interval, in addition to other criteria of the long QT syndrome, was observed in both patients. We suggest that one normal measurement of the QT interval should not rule out this diagnosis. PMID- 8225665 TI - Spontaneous hemarthrosis following thrombolytic therapy for acute myocardial infarction. AB - We describe a 45-year-old man who developed a spontaneous hemarthrosis of his right knee following thrombolytic therapy with streptokinase and rtPA for acute myocardial infarction. Surprisingly, despite the wide use of thrombolytic therapy, only four cases of spontaneous hemarthrosis following thrombolysis have been previously reported. Prompt aspiration of the joint, after stopping anticoagulant therapy, and splinting will provide early diagnosis and may prevent further damage to the joint. PMID- 8225666 TI - Streptokinase therapy in a five-year-old child with prosthetic mitral valve thrombosis. AB - Intravenous streptokinase therapy for prosthetic valve thrombosis in a 5-year-old child is reported for its rarity. The therapy is safe and effective and as in adults, should be considered the first line of treatment for prosthetic valve thrombosis in children. PMID- 8225667 TI - A scientific approach to health care reimbursement: paying for what works. PMID- 8225668 TI - Platelet ultrastructural morphology and its relevance in essential hypertension. AB - A preliminary study of platelet ultrastructural morphology by transmission electron microscopy in patients with established essential hypertension was carried out. Platelets of hypertensive patients were found to be in various stages of 'platelet activation'. According to the degree of transformation observed, platelets were classified into six forms: discoid, pseudotubular, membranous, saccular, pseudopodical and hyaline and correlated to the severity of hypertension. The pseudopodical and hyaline types platelets were particularly observed in severely hypertensive patients, and/or those with clinical evidence of target organ damage. Platelet ultrastructural changes may form the basis of a better understanding of the aetiology and pathogenesis of essential hypertension, and may serve as a marker in assessing the extent of underlying in vivo endothelial injury and tendency to thrombotic complications. PMID- 8225669 TI - Increased plasma plasminogen activator inhibitor activity after coronary spasm. AB - To examine whether ischemic attack induced by coronary spasm changes fibrinolytic activity, we examined plasma levels of tissue-plasminogen activator antigen and plasminogen activator inhibitor activity before and after hyperventilation test in patients with variant angina and in control subjects. In 12 patients with variant angina, ischemic attack associated with ST-segment elevation on the electrocardiogram was induced by hyperventilation and plasma plasminogen activator inhibitor activity levels increased significantly 15 min after the attacks (pre, 5.1 +/- 0.8; immediately after, 5.8 +/- 1.1; and 15 min after, 7.2 +/- 0.9 IU/ml, P < 0.01). In 12 control subjects, plasminogen activator inhibitor activity levels did not change. Plasma tissue-plasminogen activator antigen levels did not change in both two groups. We conclude that coronary artery spasm increases plasma plasminogen activator inhibitor activity and that it may thereby lead to the coronary thrombus formation. PMID- 8225670 TI - Effects of acetylstrophanthidin on baroreflex sensitivity in patients with acute myocardial infarction. AB - We evaluated the effects of acetylstrophanthidin on baroreflex sensitivity in patients soon after an acute myocardial infarction. Baroreflex control of heart rate is frequently depressed after acute myocardial infarction and few data are available as to the effects of pharmacological intervention on this parameter. The reflex chronotropic response to arterial baroreceptor stimulation was assessed in 29 patients with uncomplicated acute myocardial infarction in control conditions (72-96 h after symptom onset) and 30 min after acetylstrophanthidin administration. To check for spontaneous baroreflex sensitivity variations, 24 patients with the same characteristics were evaluated at the same time intervals before and after a 10-cc bolus of saline placebo. Baroreflex sensitivity was assessed by calculating the regression line relating phenylephrine-induced increases in systolic blood pressure to the attendant changes in RR intervals. Mean baseline baroreflex sensitivity value for the whole study population was 7.4 +/- 4.5 ms/mmHg and was unchanged, 7.0 +/- 4.5 ms/mmHg, after acetylstrophanthidin (P = NS). Mean baroreflex sensitivity values were also comparable dividing patients according to the site of infarction both before and after acetylstrophanthidin. Despite the lack of difference in mean baroreflex sensitivity values between the two studies, at a post hoc analysis an inverse relation was found in the total study population between baseline baroreflex sensitivity values and their changes after acetylstrophanthidin (r = -0.62; P < 0.005). The inverse relation was also evident separately in anterior (r = -0.57; P < 0.05) and in inferior (r = -0.70; P < 0.005) myocardial infarction patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225671 TI - Long-term follow-up of global and regional left ventricular function by two dimensional echocardiography after thrombolytic therapy in acute myocardial infarction. AB - In order to evaluate changes in left ventricular volumes and regional left ventricular function after thrombolytic therapy in acute myocardial infarction serial two-dimensional echocardiography was performed during a follow-up of 2 years in 206 consecutive patients treated with streptokinase and adjunctive angioplasty in a randomized group of patients. Unexpected progressive left ventricular enlargement was detected both with and without angioplasty. In anterior wall infarction, end-diastolic volume index increased from 55 +/- 14 to 91 +/- 28 ml/m2 (+65%, P < 0.01) and end-systolic volume index increased from 31 +/- 11 to 55 +/- 23 ml/m2 (+79%, P < 0.01), whereas ejection fraction decreased from 45 +/- 9 to 41 +/- 7% (-9%, P = NS). Averaged regional anterior wall motion improved during the first 4 weeks (11 +/- 10 to 16 +/- 12%), but subsequently deteriorated (16 +/- 12 to 10 +/- 6, P < 0.05). The number of segments with pathological wall motion increased. Similar volumetric and regional wall motion data were demonstrated in inferior wall infarction. We believe this reflects a chronic ventricular remodelling phenomenon. This process takes place predominantly during the first 3 months, but continues over the whole follow-up period. Forty percent of the patients suffered symptoms of heart failure on long term follow-up. Attenuation of progressive ventricular enlargement remains a therapeutic challenge in the long-term care of these patients. Angiotensin converting enzyme inhibitors are promising agents in this regard. PMID- 8225672 TI - Clinical course and left ventricular function in patients with acute myocardial infarction following delayed recanalization of infarct-related artery. AB - The effect of delayed recanalisation of the infarct related artery on clinical course and left ventricular functions was studied in 23 patients with acute myocardial infarction (Group 2). Another 82 patients with acute myocardial infarction served as controls: 48 patients with immediate recanalisation of the infarct related artery following intracoronary thrombolytic therapy (Group 1) and 34 patients with unsuccessful thrombolysis with the artery remaining occluded on repeat angiography (Group 3). Baseline clinical characteristics and left ventricular ejection fractions in the three groups did not differ statistically. Following intracoronary thrombolysis the clinical features and left ventricular functions in the three groups, respectively were as follows: post infarction angina--45.8%, 13.1% and 11.8% (1 vs. 2 and 3, P < 0.05); reinfarction--29.2%, 8.7% and 11.8% (1 vs. 2 and 3, P < 0.05); mortality--0%, 0% and 11.8% (1 and 2 vs. 3; P < 0.05); aneurysm--16.7%, 21.7% and 52.9% (1 and 2 vs. 3, P < 0.05); heart failure--20.8%, 21.7% and 47.1% (1 and 2 vs. 3, P < 0.05). Left ventricular ejection fractions on the second and on days 10-14 were, respectively, 47.6 +/- 1.1%, 42.8 +/- 1.1% and 39.2 +/- 1.6% (1 vs. 2 and 3, P < 0.05) and --52.1 +/- 1.0%, 48.9 +/- 1.1% and 44.3 +/- 1.5% (1 and 2 vs. 3, P < 0.05). Thus following delayed recanalisation of the infarct related artery the clinical course and left ventricular function improved significantly in comparison to patients without recanalisation. PMID- 8225673 TI - Reversible left ventricular dysfunction secondary to rapid atrial fibrillation. AB - We present the cases of four patients with reversible left ventricular dysfunction associated with severe congestive heart failure presumably induced by rapid atrial fibrillation. The mean heart rate was 159 beats/min and the mean left ventricular end-diastolic dimension was 58.5 mm with diffusely impaired left ventricular motion. None of the patients had a history of preceding upper respiratory infection before the acute episode and no signs of inflammation at onset, and all patients were New York Heart Association Class I or II before the acute episode. Left ventriculography, done about 1 month when congestive heart failure and ventricular rate were controlled with digitalis and diuretics, still showed diffusely decreased left ventricular motion; the mean end-diastolic volume was 165 ml and the mean ejection fraction was 30%. Coronary angiography was normal in three patients and one showed moderate left anterior descending artery stenosis. Right ventricular biopsy, done in three patients showed no evidence of myocarditis. Left ventricular wall motion normalized in 5-36 months on follow-up echocardiography. These findings suggest that persistent rapid atrial fibrillation can cause reversible left ventricular dysfunction which can take a considerable period of time to normalize. PMID- 8225674 TI - An audit of emergency echocardiography in a district general hospital. AB - Eighty patients (43 M, 37 F), aged 23-89 years who were referred for emergency echocardiography over a 12-month period were prospectively studied in order to determine the reasons for emergency echocardiography and the influence of its results on patient management. The most frequent emergency request was to clarify whether the basis for cardiomegaly in a haemodynamically unstable patient was pericardial effusion or left ventricular dilatation. Other reasons for requests were for assessment for source of systemic emboli, acute complications of myocardial infarction, endocarditis, valve dysfunction and cardiac trauma. As a consequence of the emergency echocardiography, management was immediately influenced in 19 patients. This study has provided information on the specific settings in which emergency echocardiography can be justified. PMID- 8225675 TI - Abnormal renal function in isolated systolic hypertension correlation with ambulatory blood pressure. AB - Does ambulatory blood pressure correlate with the renal function damage better than clinic blood pressure in isolated systolic hypertension, as has been reported in other target organ involvement in combined systolic/diastolic hypertension? We investigated the correlation of serum beta-2 microglobulin concentration and both 24-h urine beta-2 microglobulin concentration and excretion as measures (suggestive) of glomerular filtration rate and tubular reabsorption, respectively with both ambulatory and clinic blood pressure in 19 health normotensive (68 +/- 4.9 years) and 50 isolated systolic hypertensive elderly individuals (69 +/- 5.4 years). Serum beta-2 microglobulin concentration and 24-h urine beta-2 microglobulin concentration and excretion were higher in the isolated systolic hypertension than in the normotensive group (P < 0.05). In isolated systolic hypertensive patients, 24-h urine beta-2 microglobulin concentration and excretion were related to ambulatory blood pressure (r = 0.32 0.40, P < 0.05), but not to clinic blood pressure; waking systolic blood pressure had the strongest correlation with both 24-h urine beta-2 microglobulin concentration and excretion among derivatives of ambulatory blood pressure (r = 0.35 and 0.40, P < 0.05). We conclude that ambulatory blood pressure, especially waking systolic blood pressure, is superior to clinical blood pressure in predicting renal function impairment, in isolated systolic hypertensive patients. PMID- 8225676 TI - Development of the collagen network of the human fetal myocardium: an immunohistochemical study. AB - We have studied the development of the collagen network within the ventricular myocardium of sixteen human fetuses of gestational age 12 weeks to 22 weeks. Using the technique of indirect immunohistochemistry with antibodies raised against collagen Types I and III, we have demonstrated that the 12 week old heart has a rudimentary endomysial collagen structure which coexpresses both collagen types. Perimysial structures evolve with the onset of the second trimester and are collagen III positive before expressing collagen Type I. No differences in collagen deposition were detected between the right and left ventricular free walls, but collagen content of the interventricular septum appeared relatively high and expressed in thick highly organised fibrils. Transventricular gradients of collagen distribution were seen for both collagen isoforms which persisted with increased age. Intraventricular differences in collagen deposition were marked due to the insertion of the atrioventricular valves and tension apparatus. These findings suggest that the collagen network of the fetal myocardium is composed of copolymer fibrils, possibly rich in collagen type III which evolve at a time when the mechanical efficiency of the fetal heart must improve to keep pace with the escalating demands of a rapidly growing body. PMID- 8225677 TI - Concurrent percutaneous balloon valvuloplasty. PMID- 8225678 TI - Stokes Adams attack as the first manifestation of acute rheumatic carditis. PMID- 8225679 TI - Cutaneous and ocular changes associated with the use of chlorpromazine. AB - A 57-year-old schizoaffective disorder patient was placed on chlorpromazine because of its sedative properties and low profile for extrapyramidal side effects. After two years of treatment, the patient developed photosensitivity and blue-gray pigmentation of the skin of the face, neck and dorsum of hands. Significant pigment deposits were also observed in eye lens and cornea. An oval translucent lesion lateral to the left angle of the mouth was removed and the biopsy showed a basal cell carcinoma. The cutaneous and eye pigmentation were only partially reversible after the discontinuation of chlorpromazine. There has been no recurrence of basal cell carcinoma. PMID- 8225680 TI - Steady-state pharmacokinetics of phentermine extended-release capsules. AB - Twenty-one healthy, caucasian, male volunteers completed this randomized single blind, multiple-dose, crossover bioavailability study during which either phentermine HCl capsules (Minobese Forte, reference product) or phentermine base capsules (Duromine, test product) were ingested once daily for 14 days. A washout period of 14 days was allowed between the two treatment phases. On profile days (day 14 of each treatment phase) subjects remained recumbent for 24 hours after drug administration. Serial venous blood samples were drawn over the 24 hour dosing interval for plasma phentermine assay by gas chromatography. The 90% confidence intervals for the "test/reference" mean ratios of the pharmacokinetic variables Cmax,norm, Cmin,norm, AUCnorm (normalized for difference in the dose of phentermine base), %PTF and T75% Cmax, all fell within the bioequivalence range of 80% to 125%. With the aid of trough plasma phentermine concentrations, it was established that steady-state was reached after 14 days of once daily administration of either product. Adverse events experienced on both treatments included prolonged or recurrent episodes of insomnia, nausea, headache, dry mouth and dizziness. No clinically relevant changes in clinical chemistry or hematology variables occurred during the study. PMID- 8225681 TI - Pharmacokinetics of intravenous ataprost alfadex, a new prostaglandin I2 analog in healthy volunteers. AB - Ataprost alfadex [5(E),-6,9-deoxa-6,9 alpha-methylene-15-cyclopentyl 16,17,18,19,20-pentanor- PGI2 alpha-cyclodextrin clathrate] is a novel PGI2 derivative. From animal studies, it is expected to be a more potent inhibitor of platelet aggregation and less hypotensive than PGI2. The pharmacokinetics of ataprost were studied in 9 healthy male volunteers during and after i.v. infusion for 2 hours at the rates of 2.5 (n = 5) and 10 ng/kg/min (n = 4). Both treatments were well tolerated by the subjects. At the end of the infusion, plasma levels of 191 +/- 76 (mean +/- SD) and 645 +/- 191 pg/ml were reached, declining rapidly with half-lives of 6.7 +/- 3.0 and 5.5 +/- 0.84 minutes at the lower and higher infusion rates, respectively. The area under the plasma concentration-time curve extrapolated to infinity increased with the dose as follows: 28.7 +/- 9.8 and 80.9 +/- 24.8 ng.min/ml. The unchanged drug was not detected in urine but a metabolite was recovered in it, reaching up to 6.0 +/- 0.85% of the total dose within the first 24 hours, the most part of which was recovered within the first 4 hours. PMID- 8225682 TI - Efficacy of misoprostol in controlling indomethacin induced fecal blood loss in arthritic patients. AB - Indomethacin, a nonsteroidal anti-inflammatory drug, may cause gastric mucosal damage as shown by fecal blood loss. A randomized, double-blind, placebo controlled, parallel group study was conducted to determine the effects of 400 mcg b.i.d. misoprostol, a synthetic prostaglandin E1 analog, on intestinal blood loss caused by 50 mg t.i.d. indomethacin. Forty-two arthritic patients, mean age 59 years, received indomethacin for 14 days. Those with baseline blood loss of at least 1.5 ml/day during the first 7 days were randomized to 400 mcg of misoprostol or placebo (days 8 to 14). Fecal blood loss was measured using 51Cr labelled red blood cell technique. Success was defined as a reduction in mean daily blood loss of at least 50% during the treatment period compared to mean daily blood loss during the baseline (pre-treatment) phase. The mean daily blood loss on treatment days 9-15 was not significantly reduced from baseline in either group. These data neither confirm nor deny the effectiveness of misoprostol in reducing fecal blood loss caused by indomethacin. The results may have been confounded by the administration of misoprostol twice daily while indomethacin was administered three times daily. In addition, fecal blood loss as an indicator of gastrointestinal mucosal damage is not a sensitive measure; it is characterized by poor reproducibility and wide fluctuations within individual responses. Inappropriate laboratory techniques may have further reduced the sensitivity and reliability of this procedure. PMID- 8225683 TI - Covalent coupling of nucleosides to low density lipoprotein (LDL) generates macrophage specific (drug)-carriers. AB - Covalent coupling of thymidine and azidothymidine (ZIDOVUDINE) to epsilon-amino groups of lysine side chains of LDL-Apo B is described. This procedure generates LDL-nucleoside particles that exhibits affinity solely for the scavenger receptor pathway on macrophages which is demonstrated by cell culture experiments. Autoradiography shows that 3H-thymidine, as representative of nucleosides, is delivered to the cell nucleus. Hence internalization, lysosomal cleavage and triphosphorylation of thymidine evidently had occurred. The application of this new method of drug targeting is macrophage selective inhibition of HIV-reverse transcriptase in AIDS. PMID- 8225684 TI - A comparative bioavailability study on two sustained-release formulations of diclofenac sodium following a single dose administration. AB - A single dose comparative bioavailability study and an in vitro evaluation were conducted on two sustained-release formulations of diclofenac sodium (Voltaren "V" and Diclogesic "D"). The two products were found similar in weight and content uniformity. The in vitro dissolution, however, revealed that product D has a significantly faster rate of drug release compared to product V. The bioavailability study was carried out on 15 healthy male volunteers, who received a single oral dose (100 mg tablet) of each product according to a randomized crossover design. Blood samples were obtained over a 12 h period, and drug concentrations were determined by an HPLC assay. The two products were not found to be statistically different with respect to the lag time between dosing and appearance of the drug in the serum (0.8 +/- 0.2 and 0.6 +/- 0.1 h for V and D, respectively), or in the time needed to attain the peak concentrations (4.5 +/- 0.8 and 4.1 +/- 0.9 h for V and D, respectively). The two products, however, varied in the peak serum concentration (736 +/- 125 and 536 +/- 63 ng.ml-1 for V and D, respectively), but this difference was not statistically significance. In terms of the extent of absorption, assessed by estimating the area under the concentration-time curve over 12 h, the two products were not significantly different (3,340 +/- 270 and 3,045 +/- 294 ng.h.ml-1 for V and D, respectively). These in vitro and in vivo findings indicate that Diclogesic is characterized by sustained-release properties which are comparable to Voltaren. PMID- 8225685 TI - Dextromethorphan O-demethylation and dextrorphan glucuronidation in a French population. AB - The dextromethorphan (DMP) O-demethylation and the dextrorphan (DRP) glucuronidation distributions were studied in 120 French Caucasian subjects. After a single 25 mg DMP oral administration, DMP, free and total DRP concentrations were measured in 8h-urine collection, using an HPLC technique with fluorescent detection. The DMP and free DRP concentrations ratio, in log form, was used to estimate the oxidative demethylation phenotype of the subjects. Two different populations were found. The first one consisted of the extensive metabolizers (90.8%, 95% confidence interval from 85.6 to 95.9%) and the second one consisted of poor metabolizers (9.2%, 95% confidence interval from 4.0 to 14.4%). The antimode value of the distribution was estimated at approximately 0.7 corresponding to a ratio of 5. Moreover, this ratio was compared to the DMP and total DRP concentrations ratio, usually defined to DMP O-demethylation phenotyping. On the other hand, the glucuronide DRP concentration was calculated by subtracting the free DRP concentration from the total DRP concentration. Consequently, the free DRP and glucuronide DRP concentrations ratio was also used to estimate the DRP glucuronidation in the present population. This ratio in log form reflected the UDP-glucuronyltransferase(s) capacity(ies). This log ratio appeared to be normally distributed in the population studied. These results show that log DMP/free DRP ratio can be used, as well as log DMP/total DRP ratio, to determine the oxidative phenotype of subjects and that the DRP conjugation does not exhibit any apparent genetic polymorphism. PMID- 8225686 TI - Drug-prescription in the five new federal lands--a comparison with the prescription of the old federal lands of Germany. AB - In the present study, the consumption of several drugs on the territories of the former GDR and FRG was compared using 200,000 prescriptions and the defined daily doses (DDD). Furthermore, for the two parts of Germany, the 20 most frequently prescribed drugs were evaluated. Calcium antagonists and the beta-blockers prevailed in East Germany, while cardiac glycosides, vasodilators, reserpin containing antihypertensives, bronchodilators (beta-adrenoceptor agonists), neuroleptics, tricyclic antidepressants, antacids, H2-receptor blockers, chondroprotectives were used 1.5 to 5 times more frequently in West Germany. Because in East Germany, a shortage of drugs was no longer observed in 1991, several other reasons for the "over-prescription" of drugs for the patients in West Germany must be mentioned. PMID- 8225687 TI - Effects of dopaminergic antagonists on dopamine-induced cardiovascular and insulin secretion actions in hypertensive patients. AB - A comparative study between two dopaminergic antagonists: metoclopramide and domperidone, was undertaken in nineteen (19) hypertensive patients at the Vargas Hospital, Caracas. The patients were pretreated with labetalol, 800-1,200 mg/day, orally, over a period of one week, after which they were divided into two groups: group A, a total of eleven patients were intravenously infused with dopamine hydrochloride 0.5-3 micrograms/kg/min, before and after treatment with metoclopramide (10 mg, i.v. as a bolus); group B (n = 8), was pretreated with domperidone, 20 mg b.i.d., p.o. over a period of one week and intravenously infused with dopamine hydrochloride, 0.5-3 micrograms/kg/min. In group A, dopamine induced a decrease of blood pressure from 171.9 +/- 6.35/103.6 +/- 3.12 to 152.7 +/- 7.55/93.8 +/- 2.97 mmHg (p < 0.001) without altering heart rate, and it increased plasma insulin levels from 8.29 +/- 0.70 microunits/ml to 12.09 +/- 1.83 microunits/ml (p < 0.01). Metoclopramide caused no changes of blood pressure or plasma insulin levels. However, hypotensive responses and plasma insulin rises due to dopamine were blocked by metoclopramide. In group B, domperidone also blocked dopamine-induced antihypertensive effect (from 170.0 +/- 9.23/102.8 +/- 3.80 to 160.2 +/- 9.84/95.5 +/- 2.50 mmHg) although it was less effective than metoclopramide. Domperidone also blocked dopamine-induced increase of plasma insulin levels from 9.65 +/- 4.50 microunits/ml to 11.78 microunits/ml. We conclude that a dopaminergic receptor may be involved in some cardiovascular responses and in modulating insulin secretion in man. PMID- 8225688 TI - Potential pharmacokinetic interactions of nocloprost clathrate with retarded theophylline and enteric coated diclofenac after single and repeated premedication in healthy volunteers. AB - Pharmacokinetic interactions of cytoprotective prostaglandin E2 analog nocloprost clathrate with theophylline and diclofenac were studied in two placebo controlled, single-blind studies with parallel groups (n = 8) in healthy male volunteers (age 20-32 years, body weight 63-95 kg, body height 169-193 cm, Broca index 0.81-1.18). Nocloprost (200 micrograms) or placebo tablets were given twice daily (07:00 h a.m. and p.m.) for 8 days. Thirty min after the first administration on the first day (single) and 30 min after the last administration in the morning of the 8th day (repeated premedication), pharmacokinetic examinations with retarded theophylline capsules (250 mg) or enteric coated tablets of diclofenac (50 mg) were performed. Theophylline was measured using an HPLC- and diclofenac with a GC-method. Both single and repeated premedication with nocloprost did not significantly change any pharmacokinetic parameter of theophylline. There was only a tendency towards lower AUC of theophylline after both regimens of premedication. As far as diclofenac is concerned, single premedication increased significantly the rate of absorption and total body clearance but lowered the AUC of the NSAID. After repeated premedication, no parameter was significantly influenced. All pharmacokinetic changes observed are most likely without any clinical importance. PMID- 8225689 TI - Transdermal nicotine substitution: pharmacokinetics of nicotine and cotinine. AB - The use of tobacco has resulted in major health related problems worldwide. Nicotine replacement is one of the most promising strategies in smoking cessation. The in vivo delivery rate and the pharmacokinetic properties of a recently developed transdermal nicotine system (TNS) was investigated in three separate studies. Two sizes (16 cm2 and 24 cm2) of the patches were tested. Mean daily nicotine delivery, expressed by a first order kinetic, varied between 0.9 and 1.0 mg/cm2 patch. The different sizes of the patches led to linearly dose related nicotine plasma Cmax- and AUC-values. The nicotine levels achieved were in the same range with those of other nicotine patches and the chewing gum. As a result of multiple dose kinetic, no significant accumulation of nicotine was observed. Mean elimination half-life of nicotine after removal of the patches (3.5 +/- 1.8 h for the larger patch and 4.5 +/- 1.9 h for the smaller patch) was longer than reported values after i.v. administration, which was reported to vary between 40 and 120 min. PMID- 8225690 TI - Design and pharmacodynamic evaluation of novel dual release formulations of triazolam. AB - Triazolam is an effective hypnotic that can cause amnesia and psychomotor performance decrements, particularly after a 0.5 mg dose. Previous pharmacodynamic studies suggested a relationship between these effects and triazolam plasma concentration. A novel dual release bilayer tablet was designed to mimic the onset of action of a 0.25 mg dose and to maintain the duration of a 0.5 mg dose without the side effects associated with the 0.5 mg dose. The immediate release component of the bilayer tablet contained 0.25 mg triazolam while the sustained release component contained 0.15 mg triazolam. Two prototype formulations of the bilayer tablet, differing in rate of release in the sustained release component, were tested against a conventional 0.5 mg triazolam compressed tablet and placebo in a single-dose, double-blind, four-way crossover study in healthy male subjects. Triazolam plasma concentration time profile was obtained over 12 hours following single administration of each treatment. Effects of triazolam on central nervous system function were evaluated using psychomotor performance tests, immediate and delayed recall tests and rating of sedation. The triazolam plasma concentrations were not significantly different among the active drug treatments, although the dual release tablets did give the expected profiles. There were significant differences in triazolam effects on memory and psychomotor performance. The slowest releasing dual-release tablet showed significantly less psychomotor impairment and memory deficit than the conventional tablet. There was no difference in sedation among the active drug treatments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225691 TI - Qualitative analysis of drug prescription in a geriatric population sample. AB - The rationality of specialist drug prescription in 154 patients older than 64 in 3 Zagreb homes for the aged was analyzed over an 11-month period. Categories analyzed were: number of prescribed drugs, justification of prescription, adequacy of the dose, route, form and therapy duration of the prescribed drugs, adequacy of the type of the prescribed drugs in the advanced age and the presence of contraindications of drugs used. The criterion of the rational drug use was the WHO's modified definition of rational drug therapy: "Application of an appropriate drug by a correct route in an adequate dose over a sufficiently long period of time". This basic criterion was further elaborated in relation to the analyzed categories. There were 2.66 (+/- 1.65) prescribed drugs per visit in which drugs were prescribed. In all analyzed categories, distinctive aberrations from the principles of rational drug prescription were found. One hundred and sixty-four (41.4%) of all drugs were unjustifiably prescribed and 103 (26.4%) were not dosed correctly. The duration of the therapy was inadequate for 60 (15.4%), route for 41 (10.5%), form for 32 (8.2%) of drugs prescribed. Seventy five (19.3%) were not adequate because of patients' age and 15 (3.9%) were prescribed in spite of existing contraindications. PMID- 8225692 TI - Assessing the risk of ovulation in interaction studies of drugs and oral contraceptives. AB - When certain drugs are taken concomitantly with an oral contraceptive, the efficacy of the contraceptive may be obtunded, and pregnancy may ensue. Since the function of oral contraceptives is to suppress ovulation, the risk of ovulation is an important parameter in clinical studies investigating interactions between drugs and oral contraceptives. The purpose of this paper is to compare a crossover design, and a new study design used for assessing a possible interaction between a drug and an oral contraceptive, and to discuss the statistical issues involved in the planning and evaluation of the results of such studies. PMID- 8225693 TI - Detection of glucose intolerance in pregnancy. AB - Current methods for the screening and diagnosis of glucose intolerance first discovered in pregnancy are reviewed and innovative approaches to the detection of metabolic disturbances in pregnancy are presented. Glucose intolerance first detected in pregnancy, termed gestational diabetes mellitus (GDM), is amongst the most significant risks of adverse fetal and maternal outcome. Normal pregnancy is characterized by both insulin resistance and pancreatic B cell compensation. In those pregnancies complicated by glucose intolerance reflected in hyperglycemia, insulin resistance appears to be heightened, both blood flow and transcapillary transport of insulin are compromised and insulin receptor and post receptor defects are exacerbated. The resulting hyperinsulinemia and hyperglycemia have, in turn, been associated with accumulated maternal fat deposition and fetal macrosomia. This cascade of events constitutes GDM or impaired glucose tolerance. The discovery of GDM is made through a process of screening and diagnosis, employing standardized oral glucose challenge tests. These tests were designed to identify those women at risk for subsequent development of non-insulin dependent diabetes mellitus. The current efficacy of glucose challenge tests has been questioned in light of increasing concern over their usefulness in detecting those women at risk for maternal and fetal complications of pregnancy. Alternative methods, including both the modification of the standardized tests, as well as the introduction of newer methodologies, such as capillary blood glucose monitoring, have been proposed. The implementation of newer approaches may result in improved detection of those women whose infants are at high risk for both metabolic and morphologic complications of persistent hyperglycemia in pregnancy. PMID- 8225694 TI - Cocaine: friend or foe? (Part 1). AB - The history of cocaine use goes back a long time. Initially, cocaine was used as a medicine by physicians and as a pleasure agent in over-the-counter preparations. The significance and intensity of cocaine use and abuse became well known soon thereafter and resulted in extensive regulations to curb its use. Cocaine is a powerful anesthetic agent with powerful vasoconstrictive properties. It can be absorbed from all mucous membrane sites and results in severe cardiovascular complications. The effects of cocaine on various organ systems are presented here. PMID- 8225695 TI - A pilot study on the liver protective effect of silybin-phosphatidylcholine complex (IdB1016) in chronic active hepatitis. AB - In order to assess the liver protective activity and the antioxidant properties of a new silybin complex (IdB1016), we carried out a short-term pilot study on 20 patients with chronic active hepatitis (CAH), randomly assigned to 240 mg of silybin b.i.d. (10 patients, 4 m/6 f, mean age: 50 years) or placebo (10 patients, 2 m/8 f, mean age: 55 years). Blood samples were collected before and after 7 days of treatment for liver function tests (LFTs), malonaldehyde (MDA) as an index of lipid peroxidation, and copper (Cu) and zinc (Zn), two trace elements involved in protecting cells against free radical-mediated lipid peroxidation. In the treated group, there was a statistically significant reduction of mean (+/- SEM) serum concentrations of aspartate aminotransferase (AST) from 88.0 (+/- 13.3) to 65.9 (+/- 7.5) u/l, (p < 0.01), of alanine aminotransferase (ALT) from 115.9 (+/- 12.9) to 82.5 (+/- 10.6) u/l (p < 0.01), of gamma glutamyltranspeptidase (gamma-GT) from 51.4 (+/- 9.3) to 41.3 (+/- 4.2) u/l (p < 0.02) and of total bilirubin (TB) from 0.76 (+/- 0.08) to 0.53 (+/- 0.04) mg/dl (p < 0.05). Alkaline phosphatase (AP) fell slightly from 143.4 (+/- 6.4) to 137.5 (+/- 7.8) u/l. There were no significant changes in MDA, Cu or Zn serum concentrations. These results show that IdB1016 may improve LFTs related to hepatocellular necrosis and/or increases membrane permeability in patients affected by CAH. PMID- 8225696 TI - Lack of effect of cimetidine on furosemide kinetics and dynamics in patients with hepatic cirrhosis. AB - The influence of cimetidine on the natriuretic and diuretic responses to furosemide was studied in 10 patients with hepatic cirrhosis. After four days on a low sodium diet, the patients were given 40 mg of furosemide i.v. and from the sixth to the eleventh day they received 400 mg of cimetidine p.o. every 6 h and a second dose of furosemide with the last 6 h dose. Ten healthy subjects received the same dose of furosemide. Multiple blood and urine samples from both groups were analyzed for furosemide, sodium and creatinine. Furosemide kinetics were not affected in patients with hepatic cirrhosis but the effect was lower than in the controls: urinary excretion of sodium (0.47 +/- 0.07 vs 1.59 +/- 0.10 mmol/min, p < 0.05) and urine excretion (4.86 +/- 0.57 vs 9.16 +/- 0.85 ml/min, p < 0.05). The predicted maximal effect of furosemide (Emax) and the furosemide urinary rate of excretion needed to elicit 50% of Emax for the cirrhotic patients and the controls were 1.85 +/- 0.21 and 3.22 +/- 0.41 mmol/min (p < 0.05) and 137 +/- 15 and 99 +/- 12 micrograms/min (p < 0.05), respectively. The amounts of sodium filtered (FNa) and reabsorbed in response to the injection of furosemide were lower in the cirrhotic patients than in the controls, however, relative to the FNa, the cirrhotic patients reabsorbed more sodium than the controls. The administration of cimetidine did not affect the kinetics of furosemide nor its natriuretic or diuretic responses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225697 TI - A non-invasive evaluation of flosequinan on haemodynamics and exercise capacity in chronic congestive heart failure. AB - The efficacy of flosequinan 100 mg once daily was evaluated in 15 patients with severe congestive heart failure (New York Heart Association [NYHA] class II-IV) who had not responded adequately to digoxin and diuretics. Efficacy assessments using non-invasive techniques included exercise capacity, haemodynamics and left ventricular function. Determinations were made after 3 and 21 days' treatment, and compared with baseline. Flosequinan significantly increased exercise capacity by 27% after 3 days (+79 seconds, p = 0.015) and by 43% after 21 days (+123 seconds, p = 0.0007) and was accompanied by an increase in heart rate (+7.2 beats/min, p = 0.03; +9.1 beats/min, p = 0.03, respectively). Cardiac index and cardiac output were also significantly increased but only after 21 days' treatment (+0.3 l/min/m2, +16% and +0.5 l/min, +14%, respectively; both p = 0.008). Flosequinan was well tolerated, with headache being the most frequently reported adverse event and only 1 patient being withdrawn. One patient died but this was not unexpected in a group of patients with severe heart failure. Using non-invasive techniques this study demonstrated that in patients with severe chronic congestive heart failure, flosequinan increased exercise capacity and cardiac output, the latter being achieved mainly by an increase in heart rate. PMID- 8225698 TI - Amlodipine in ambulatory hypertensive patients: humoral and haemodynamic effects. AB - In order to evaluate the mid-term effects of amlodipine, a 1,4-dihydropyridine calcium antagonist, as well as its effects on the renin-angiotensin-aldosterone system (RAAS), on water and electrolyte balance, on urinary excretion of albumin (UAE) and on lipid metabolism, thirteen hypertensive patients (2 M, 11 F, mean age 54 years) were studied in the course of 24 weeks of therapy with amlodipine at 5-10 mg/day. Pre-therapy and periodically during therapy, the systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) were recorded in the sitting orthostatic positions (2 measurements). Laboratory tests were performed at times O and at 24 wks with the patients fasting for at least 12 h in the recumbent position. The tests included: plasma renin activity (PRA), plasma aldosterone (PA), serum angiotensin converting enzyme (SACE), blood urea nitrogen (BUN), blood creatinine, plasma electrolytes (Na, K, Cl), triglycerides, total cholesterol (TC) and HDL-cholesterol (HDLC), along with 24-h urine samples (with sterile urine) to determine UAE. The results of this study demonstrate that SBP, DBP and HR were significantly reduced during the 24 wks of therapy. The water and electrolyte and renal function were not modified. After treatment the levels of TC were significantly reduced. No change was observed in the RAAS, while the mean levels of UAE were reduced though not significantly. In conclusion, amlodipine was shown to be effective for the therapy of hypertension; it does not cause reflex tachycardia even in mid-term therapy and was effective in reducing TC levels. PMID- 8225699 TI - Biochemical analysis of rheumatoid synovial fluid after serial intra-articular injection of high molecular weight sodium hyaluronate. AB - A high-molecular-weight sodium hyaluronate (commercial name Artz) was serially injected biweekly for 12 weeks into 9 knee joints from 8 patients with active rheumatoid arthritis (RA) and certain clinical and biochemical parameters were monitored. Three of these parameters, namely specific viscosity, concentration of HA, and stringency, were significantly increased by the intra-articular injection of Artz. Clinically, the majority of the knees (7 out of 9 injected) showed a relief from pain and no joint fluid could be drawn from 4 of the knees after 12 weeks following the initial injection. Both the fluid volume collected and the molecular weight of hyaluronate in the synovial fluid showed some improvement, but it was not statistically significant. No improvement of systemic inflammatory parameters such as erythrocyte sedimentation rate and C reactive protein were observed. Injectable Artz sodium hyaluronate was effective for the treatment of local inflammation in RA. PMID- 8225700 TI - Fluvoxamine-induced alterations in plasma concentrations of imipramine and desipramine in depressed patients. AB - The effect of fluvoxamine maleate, 100 mg/day for 10 days, on plasma concentrations of tricyclic antidepressants was studied in 15 depressed patients on maintenance therapy with imipramine (7 pts.) or desipramine (8 pts.). In the subgroup treated with imipramine, plasma levels of imipramine increased significantly (p < 0.001) during fluvoxamine coadministration, while levels of desipramine were not modified. Addition of fluvoxamine to patients on a stable desipramine dosage regimen resulted in a slight, but statistically not significant, increase in desipramine plasma concentrations. These results suggest that fluvoxamine is a potent inhibitor of imipramine demethylation, while it has a weak effect on the hydroxylation of desipramine. PMID- 8225701 TI - Effects of seaprose on the rheology of bronchial mucus in patients with chronic bronchitis. A double-blind study vs placebo. AB - There are changes in the rheological characteristics of mucus (viscoelasticity) in several pulmonary pathologies, and especially in chronic bronchitis. Seaprose, a proteolytic enzyme, is one of the pharmacological possibilities for affecting the rheology of bronchial mucus to correct mucostasis and improve its clearance. The action of this drug on the viscoelasticity of bronchial mucus was assessed in a double-blind vs placebo study with 20 randomly balanced chronic bronchitis patients using a new kind of portable rheometer with special features designed for routine bronchial mucus analysis in clinical practice at the patient's bedside. It was found that in the group of patients who were given the placebo, there were no particular changes in the rheological behaviour of mucus, while in those patients who were given seaprose there were significant changes in both viscosity and elasticity at the end of treatment. Eight days after the end of treatment with seaprose, there was still a significant beneficial effect on the viscoelasticity of mucus and a sort of "post-mucolytic effect" can be postulated. Seaprose also had antiinflammatory action, and since in chronic bronchitis there are variable degrees of inflammations, its beneficial long-lasting effect could also be ascribed to this concomitant action. PMID- 8225702 TI - In vitro interaction between clindamycin and trimethoprim in combination on staphylococci. AB - The in-vitro interaction between clindamycin and trimethoprim was tested on 10 staphylococcal clinical isolates by the checkerboard technique and by the time kill curve. Indifference was demonstrated against seven of these strains and antagonism against three. The clindamycin/trimethoprim combination is of no value if the purpose of the combination is to obtain synergy against staphylococci. However, the combination is useful against mild polymicrobial infections due to Gram-positive aerobes, anaerobes and Enterobacteriaceae. PMID- 8225703 TI - The influence of L-acetylcarnitine on reinnervation of the oculomotor nerve. AB - In guinea-pigs the oral administration of L-acetylcarnitine (L-AC) markedly favours the process of reinnervation of the oculomotor nerve sectioned at intracranial level. The gains of the horizontal and vertical vestibulo-ocular reflexes (HVOR, VVOR) were taken into consideration in testing the functional recovery of the nerve. As a consequence of the drug administration, 24 weeks after the operation the gains of the treated animals were higher than those of the controls. Reduction of misalignments of the stimulus-response orientation was also observed in treated animals as compared to the controls. This suggests that L-AC potentiates motor reinnervation by enhancing the nerve-growing processes and favouring a better consolidation of the appropriate neuromuscular synapses. The increased gain, and the improvement of the alignment in ocular responses, due to L-AC would allow for an increase of visual function during head movement by optimizing gaze stability. PMID- 8225704 TI - An ergonomic approach for the evaluation of health risk in drug research and production. AB - Scientists and workers engaged in the study and production of drugs may be exposed to harmful events not yet adequately evaluated. An ergonomic approach to this problem with a view to defining projects of primary prevention, based on the respect for organizational congruences and on the knowledge of risk sources and dynamics, can represent an important measure of security and also a way to promote scientific advancement. PMID- 8225705 TI - Cutaneous lymphoma: a clinically relevant classification. PMID- 8225706 TI - Status of medical treatment for androgenetic alopecia. PMID- 8225707 TI - Ancient observations of "uncombable hair syndrome". PMID- 8225708 TI - Syphilis 100 years ago: parallels with the AIDS pandemic. PMID- 8225709 TI - Clinical, biochemical and morphologic features of acne keloidalis in a black population. AB - BACKGROUND: Acne keloidalis (AK) is an important cause of morbidity in Nigeria and accounts for 1.3% of patients with skin conditions in a Nigerian dermatology clinic. Treatment is usually unsatisfactory because the etiopathogenesis is unclear. METHODS: A prospective clinico-pathological study was carried out to identify predisposing factors, viable treatment modalities, and prognostic indicators. RESULTS: The study suggested that AK is associated with the male gender seborrheic constitution, early reproductive years, and increased fasting blood testosterone concentration. Features that may predispose to the vastly predominant occipital location of the lesions include increased mast cell density and dilatation of dermal capillaries. MANAGEMENT: The main aims are diagnosis of early papules and avoidance of physical and chemical traumatizing agents. Retinoic acid analogs and antiandrogens may be helpful. CONCLUSIONS: The widespread use of irritating physical and chemical traditional treatment remedies and delay in seeking medical attention--AK is typically asymptomatic--contribute to the relatively advanced nature of the disease at the time of presentation to the specialist. PMID- 8225710 TI - Non-melanoma skin cancer and keratoacanthoma in Filipinos: an incidence report from Kauai, Hawaii. AB - BACKGROUND: Non-melanoma skin cancer is the most common malignancy in the white population of the United States with an estimated 700,000 new cases each year. Regrettably, data on minority racial groups are either scarce or lacking entirely. METHODS: This study was designed as a 5-year prospective incidence study of non-melanoma skin cancer and keratoacanthoma by using an island-wide survey of Kauai's Filipino residents and covers the years of 1983 to 1987. RESULTS: Seven basal cell carcinoma (incidence: 12.3/100,000), one squamous cell carcinoma (incidence: 1.8/100,000) and four keratoacanthoma (incidence: 7/100,000) patients are reported. CONCLUSIONS: To the best of our knowledge, this is the first population-based incidence report on non-melanoma skin cancer and keratoacanthoma in this population. PMID- 8225711 TI - Risk of cancer in relatives of patients with cutaneous melanoma. AB - BACKGROUND: Cutaneous malignant melanoma (CMM) is a recognized feature of the Lynch type II cancer-family syndrome and the Li-Fraumeni's syndrome. A significant contribution of these syndromes to the total burden of CMM would be reflected in an increased risk of nonmelanoma cancers in first degree relatives. METHODS: Pedigrees were taken from 85 patients with CMM using a family history questionnaire. The relative risk of death from all cancers and individual cancers in first degree relatives was calculated. RESULTS: Of the 85 questionnaires, those of 79 patients were completed and of adequate quality for analysis. The first degree relatives of CMM patients showed no increased risk of cancer death, the relative risk of cancer death being 1.0. Six patients (7.6%) had first degree relatives with CMM. One patient had a family history compatible with the dominant transmission of a predisposition to cancer. CONCLUSIONS: It is important to establish whether an increased cancer risk is present in relatives of patients with malignancies so that screening programs may be offered. This study provides little evidence to support seeing relatives for noncutaneous malignancies in the absence of a dominant family history of predisposition to cancers. The increased frequency of CMM in relatives suggests that relatives of CMM patients should be counseled on protection from the sun and examination of the skin for melanoma. PMID- 8225712 TI - Effective detection of plantar malignant melanoma. AB - BACKGROUND: As the sole of the foot is the most prevalent site of malignant melanoma in non-Caucasians, early detection of the neoplasm at this anatomical site is very important. In our previous study, we proposed a clinical guideline that acquired melanocytic lesions on the sole larger than 7 mm in maximum diameter should be examined histologically. METHODS: Eighty-one Japanese patients with the complaint of plantar pigmented lesions were screened at our dermatology clinic during 3 years using the 7-mm criterion. RESULTS: Of the total 80 melanocytic lesions on the sole, 14 lesions were larger than 7 mm in maximum diameter, excluding congenital lesions. Diagnoses of the 14 "large" lesions were as follows: advanced malignant melanoma, 8 lesions; early malignant melanoma (malignant melanoma in situ), 1 lesion; acquired melanocytic nevus, 4 lesions, and volar melanotic macule, 1 lesion. CONCLUSIONS: The present study confirmed the validity of the 7-mm criterion for the early effective detection of plantar malignant melanoma. PMID- 8225713 TI - Reticulate acropigmentation of Kitamura: two case reports. AB - BACKGROUND: Reticulate acropigmentation of Kitamura (RAK) is an autosomal dominantly inherited dermatosis. CASE REPORTS: Two patients are described with the clinical and histopathologic features of reticulate acropigementation of Kitamura. CONCLUSIONS: We speculate that RAK and the Dowling-Degos disease (DDD) are the same diseases. PMID- 8225714 TI - Aggressive trichoepithelioma versus keratotic basal cell carcinoma. PMID- 8225715 TI - Nail and skin hyperpigmentation associated with hydroxyurea therapy for polycythemia vera. PMID- 8225716 TI - Crusted scabies: an unusual clinical manifestation. PMID- 8225717 TI - Cutaneous B-cell lymphoma consisting of large cleaved cells with multilobated nuclei. PMID- 8225718 TI - Black dot tinea capitis in a man. PMID- 8225719 TI - Malignant melanoma in Turner's syndrome. PMID- 8225720 TI - Nevoid hyperkeratosis of the areola. PMID- 8225721 TI - Desmoplastic trichoepithelioma and multiple epidermal cysts. PMID- 8225722 TI - In situ porocarcinoma: a case with malignant expression in clear tumor cells. PMID- 8225723 TI - Urticarial vasculitis developed on the striae distensae during pregnancy. PMID- 8225724 TI - Oral mini-pulse therapy with betamethasone in vitiligo patients having extensive or fast-spreading disease. AB - BACKGROUND: Systemic corticosteroids can arrest the progression of vitiligo and lead to repigmentation in a significant proportion of patients, but may also produce unacceptable side effects. To minimize the side effects, we tried a new approach using mini-pulse therapy with betamethasone. METHODS: Forty patients having extensive and/or fast-spreading vitiligo were given 5 mg betamethasone/dexamethasone as a single oral dose after breakfast on 2 consecutive days per week. The response to treatment was evaluated by photographs taken every 2-4 months and recording the side effects. RESULTS: Within 1-3 months, progression of the disease was arrested in 89% of the 36 patients having active disease, while 2 patients needed an increase in the dose to 7.5 mg per day to achieve complete arrest of lesions. Within 2-4 months, 80% of the patients started having spontaneous repigmentation of the existing lesions which progressed with continued treatment. The extent of repigmentation varied in different patients and even in different lesions in the same patient. It was less than 10% in 14 (35%) patients and almost complete (> 90%) in three patients. The side effects included weight gain of 5 and 7 kg in two patients, mild headache in two patients, transitory general weakness for 2 days after the pulse in two patients, and bad taste in the mouth in three patients; 23 patients, including six children, had no side effects. CONCLUSIONS: Oral mini-pulse therapy with betamethasone/dexamethasone seems to be an effective treatment modality to arrest the progression of vitiligo. It also induces spontaneous repigmentation. It deserves to be tried on a large scale to evaluate its advantages over the currently available methods of treatment. PMID- 8225725 TI - Use of topical minoxidil therapy for androgenetic alopecia in women. AB - BACKGROUND: Androgenetic alopecia is the most common cause of hair loss in men and women. Androgenetic alopecia in women begins as a diffuse and progressive thinning of the frontoparietal area of the scalp. In women, hair loss at any age is socially unacceptable and may be the basis of psychiatric illness. METHODS: A 32-week, double-blind, placebo-controlled trial was conducted in 10 European centers to assess the efficacy and safety of 2% topical minoxidil solution for the treatment of androgenetic alopecia in women. Two hundred ninety-four of the 346 women enrolled (85%) completed the 32-week trial. Photographic and computer imaging techniques were used at each visit to determine objectively the number of nonvellus hairs present in a 1-cm2 area selected as the target evaluation site. RESULTS: In the 2% minoxidil group, the mean increase in nonvellus hair count was 33 hairs, which was significantly greater than that of 19 hairs in the placebo group (P = 0.0001). The investigators observed that 44% of the patients in the 2% minoxidil group achieved new hair growth compared with 29% in the placebo group. When asked to evaluate their own hair growth, 55% of the women in the 2% minoxidil group compared to 41% of the women in the placebo group believed that they had achieved new hair growth. No clinically significant changes in vital signs were observed during the study and no serious or unexpected medical events were reported. CONCLUSION: Topical minoxidil solution was significantly more effective than placebo in the treatment of androgenetic alopecia in women. PMID- 8225726 TI - Topical minoxidil therapy for androgenic alopecia in the Middle East. The Middle Eastern Topical Minoxidil Study Group. AB - BACKGROUND: A 48-week open label trial was conducted in five Middle-Eastern countries (Lebanon, Egypt, Saudi Arabia, Jordan, and the United Arab Emirates) to determine the safety and efficacy of 2% minoxidil in the treatment of early androgenic alopecia and to compare the response with Western countries. METHODS: One hundred and ninety-five men aged between 19 and 47 years were enrolled. The duration of their baldness varied from 6 months to 10 years, and they all showed a type III vertex or type IV of the modified Hamilton scale. Baldness pattern, diameter of the balding area, hair counting within a 2.5 cm bald patch as well as investigator's and patient's evaluation were regularly noted. RESULTS: No significant changes were observed in vital signs or laboratory parameters. Of the 161 patients considered evaluable at 48 weeks, 80% showed moderate to dense growth. The mean increase in nonvellus hair at 12 months was 234. CONCLUSIONS: The age of the patient and the type of baldness rather than its duration affected the final outcome. PMID- 8225727 TI - Marriage: a 19th century French method for the prevention of syphilis: reflections on the control of AIDS. PMID- 8225728 TI - Cancer mortality in a cohort of rural licensed pesticide users in the province of Rome. AB - The mortality of a cohort of 2310 male workers who obtained a licence to handle pesticides in the period 1973-1979 in the province of Rome was investigated. The cohort contributed 26,846 person-years of exposure. The vital status of the cohort was determined up to the end of 1988. The causes of death of the 207 who had died were ascertained from death certificates. Standardized mortality ratios (SMR) were computed using both the provincial and the national mortality rates. The two methods yielded very similar results. Using the national rates, the SMR for all causes was 56 (95% confidence interval (CI) : 45.3-59.8), for cardiovascular diseases 47 (95% CI : 37.1-59.1), and for all cancers 72 (95% CI : 57.8-89.3). A statistically significant excess was noted for brain cancer (SMR = 270, 95% CI : 108.6-556.9). In addition, the cohort experienced statistically significant lower lung cancer mortality (SMR = 57, 95% CI : 35.6-80.0). PMID- 8225729 TI - An historical cohort study of cardiac catheterization during childhood and the risk of cancer. AB - This study aimed to determine whether cancer risk was elevated among patients exposed to radiation from diagnostic cardiac catheterization during childhood. The study cohort included 3915 children who underwent at least one cardiac catheterization at a major children's hospital in Toronto, Canada, between 1950 and 1965, were < or = 18 years at the time of the procedure, and were residents of Ontario. Follow-up of the cohort was done by linkage to the Ontario Cancer Registry up to 1986. A total of seven cancer deaths were observed (O), compared with 5.7 expected (E) based on provincial cancer rates (mortality ratio [O/E] = 1.2; 90% confidence interval [CI] : 0.6-2.3). In the analysis of cancer incidence, 13 cancers were detected, compared with 17.3 expected (O/E = 0.75; 90% CI : 0.4-1.2). These mortality and incidence ratios were not statistically significantly elevated. Detailed analyses were conducted according to age at exposure, age at diagnosis, sex, number of procedures, year of first catheterization, and latent period. Detected deviations in risk were confined to an early period of exposure, indicating that later practices of cardiac catheterization were not associated with increased risk. In general, there was no statistically significant excess risk of cancer among the cohort. PMID- 8225730 TI - Race and sex differences in lung cancer risk associated with cigarette smoking. AB - Despite the extreme differences in the incidence of lung cancer between black and white Americans of each sex, little information is available on the sex-race specific lung cancer risk due to tobacco use. In the current study, case-control data were examined for sex-race differences in the lung cancer risk associated with cigarette smoking. Results indicate that Kreyberg I lung cancers (squamous cell and oat cell carcinomas) are associated with heavier intensity of smoking than Kreyberg II lung cancer (adenocarcinomas and alveolar cell carcinomas); blacks are at higher risk than whites (relative risk = 1.8), and women are at higher risk than men for a given level of smoking (RR = 1.7). Our findings indicate the existence of important differences in the smoking-associated risk for lung cancer which depend upon sex, race, and histology. PMID- 8225731 TI - Increased alcohol-related oesophageal cancer mortality rates in Japanese men. AB - Age-adjusted oesophageal cancer mortality rates for Japanese women declined by 58% between 1960 and 1989, whereas corresponding rates for Japanese men have shown no decline. We speculate that alcohol-related oesophageal cancer mortality rates have been increasing in Japanese men replacing non-alcohol related oesophageal cancer deaths. Specifically, male birth cohorts, which experienced increased alcohol-related cirrhosis mortality rates, would also experience a rise in oesophageal cancer mortality rates. To test this hypothesis, we compared male to female ratios of oesophageal cancer mortality rates by birth cohort with those of liver cirrhosis mortality rates. We calculated the attributable risk of alcohol consumption and smoking to oesophageal cancer in Japanese men using oesophageal cancer mortality rates in Japanese women as a baseline, i.e. non alcohol and non-smoking related oesophageal cancer deaths. We applied this method to head and neck cancer deaths to test its feasibility. Male birth cohorts born after 1926, which experienced male to female cirrhosis mortality ratios, also experienced increased oesophageal cancer mortality ratios. Overall, drinking and smoking accounted for 86% of all oesophageal cancer deaths and 85% of head and neck cancer deaths among Japanese men. PMID- 8225732 TI - Vodka consumption, tobacco smoking and risk of gastric cancer in Poland. AB - A multicentre hospital-based incident case-control study with 520 male gastric cancer cases aged < 75 years and an equal number of age- and sex-matched controls without cancer has been carried out in Poland to assess potential risks due to smoking and alcohol consumption. It was shown that after adjusting for socio demographic and dietary confounders and vodka drinking, smoking cigarettes had no significant effect on risk. The estimated relative risk (RR) increased to 2.27 (95% confidence interval [CI] : 0.97-5.28) for intestinal cardia cancer for those who smoked cigarettes without filters. The RR for stomach cancer grew as the frequency and amount of vodka drunk increased. People drinking vodka at least once a week had about a threefold higher risk compared to non-drinkers (RR = 3.06, 95% CI : 1.90-4.95). The effect of vodka drinking on risk was particularly strong for non-cardia cancers of the intestinal type. Those who usually drank vodka before breakfast had an elevated risk (RR = 2.98, 95% CI : 1.60-5.53) which was also present in all the subgroups investigated. Cardia and non-cardia cancer showed differences with respect to the interaction between tobacco smoking and vodka drinking. For cancers of the cardia region the risk was low for non-smokers or those who drank small amounts of vodka. The risk for cardia cancer increased considerably for smokers of cigarettes without filters and vodka drinkers who consumed large amounts (RR = 3.70, 95% CI : 1.13-12.06). For the non-cardia region a uniform increase could be observed for vodka drinking regardless of cigarette smoking status.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225733 TI - Family history of breast, ovarian and endometrial cancer and risk of breast cancer. AB - The relationship between family history of breast, ovarian and endometrial cancer and risk of breast cancer was analysed using data from a case-control study of breast cancer conducted in the greater Milan area, Northern Italy. The cases studied were 3415 women (median age 52 years, range 23-74) who had histologically confirmed breast cancer diagnosed within the year preceding the interview. The controls were 2916 women (median age 54 years; range 21-74) in hospital for a spectrum of acute illnesses excluding gynaecological, hormonal or neoplastic conditions. A total of 375 cases (11.0%) and 128 controls (4.4%) reported a history of breast cancer in first degree relatives. Compared with women with no family history of breast cancer, the RR was 2.7 (95% confidence interval [CI] : 2.2-3.3) in those with one first degree relative affected and 2.8 (95% CI : 1.3 5.7) in those with two or more affected relatives. In comparison with women without family history of ovarian cancer the RR of breast cancer was 1.4 (95% CI : 0.9-2.3) for those reporting one or more first degree relatives with ovarian cancer. However, the multivariate estimate for family history of ovarian cancer, including a term for familial breast cancer, decreased to 0.8 (95% CI : 0.5-1.4). The risk of breast cancer was similar in women reporting a family history of breast cancer (RR = 2.2) and in those reporting a family history of both breast and ovarian cancer (RR = 2.5), in comparison with women reporting no family history of breast and/or ovarian cancer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225734 TI - Breast cancer and prolonged lactation. The WHO Collaborative Study of Neoplasia and Steroid Contraceptives. AB - The possible relationship between breastfeeding and risk of breast cancer was assessed in multinational hospital-based case-control study that was conducted from 1979 to 1986 in 10 countries, including some in which many women lactated for long periods of time. Data were collected by personal interviews of 2336 cases and 14,900 controls. No statistically significant trends in relative risk estimates with duration of lactation were observed in all women combined, in women who were pre-or postmenopausal, in women of various ages at diagnosis, in women with different numbers of livebirths, or in women characterized by the presence or absence of other risk factors for breast cancer. No trends in risk were observed with mean number of months that a woman lactated per breastfed child, or with number of pregnancies followed by treatment with oral or injectable medication to suppress lactation. In premenopausal women, and in those with two or more livebirths, most risk estimates for women who lactated for various lengths of time > 6 months, relative to the risk in women who breastfed for < or = 3 months, were less than unity, but the 95% confidence intervals of all estimates included one. Long-term lactation may reduce slightly the risk of breast cancer, but the evidence for this from the present and prior investigations is not strong. PMID- 8225735 TI - Radon exposure and leukaemia in adulthood. AB - Positive associations between leukaemia and radon concentrations have been observed in England, Scotland and Wales, and Canada. Results of a similar study for the populations of 41 French administrative areas ('departements') are reported for 1984-1986. The average indoor radon and gamma ray concentrations per 'departement' range from 12 to 147 Bq.m-3 and from 28 to 142 nG.h-1, respectively. Acute lymphoid leukaemia mortality rate is similar to the national level, whereas an excess of acute myeloid leukaemia deaths is observed. According to Poisson regression models and modified tests for partial correlation, acute myeloid leukaemia mortality is significantly and positively related to indoor radon concentration whether or not adjustment is made for indoor gamma ray dose, socioeconomic status and linear gradient. This result reinforces the evidence that indoor exposure to high levels of radon is a leukaemic environmental hazard. PMID- 8225736 TI - Seasons, temperature and coronary disease. AB - Using data from a community-based register of heart disease (the WHO MONICA Project) associations between daily temperature, rainfall and other seasonal effects were investigated in relation to fatal coronary events and non-fatal definite myocardial infarctions in an Australian population. Coronary events, both fatal and non-fatal, were 20-40% more likely to occur in winter and spring than at other times of the year. Coronary deaths were more likely to occur on days of low temperature (and to a much lesser extent, on days of high temperature). No differences were found between patterns of sudden and non-sudden deaths (those occurring later after the onset of symptoms) associated with weather conditions. Statistical models allowing simultaneously for longer-term seasonal effects and daily temperature effects suggested that both exist. These results suggest that avoiding temperature stress could lead to reductions in the annual peaks in coronary events. PMID- 8225737 TI - Relationships between cigarette smoking, blood pressure and serum lipids in the Singapore general population. AB - In a cross-sectional random survey of the whole of Singapore (2143 subjects aged 18-69 years), cardiovascular risk factors were measured by standardized techniques. For the analysis in the 18-69 year age group there was adjustment for ethnic group, age, body mass index, alcohol consumption and physical activity. Among serum lipids, high density lipoprotein (HDL-) cholesterol and fasting triglyceride were inversely related with partial correlation coefficients (r) of males -0.34 (P < 0.001) and females -0.26 (P < 0.001). There were no relationships between blood pressure and serum lipids except for direct ones with fasting triglyceride, being males (systolic r = 0.06, P = 0.066 and diastolic r = 0.12, P < 0.001) and females (systolic r = 0.11, P < 0.001 and diastolic r = 0.13, P < 0.001). Cigarette smoking, in males, was related to systolic blood pressure (inversely), with, compared to non-smokers, a reduction of 1.3 mm Hg (1.1%) in light smokers, 3.8 mm Hg (3.1%) in moderate smokers and 4.6 mm Hg (3.7%) in heavy smokers; there was no clear relation with diastolic blood pressure. Cigarette smoking, in males, was related to HDL-cholesterol (inversely), even after further adjustment for fasting triglyceride, with compared to non-smokers reductions of 0.03 mmol/l (3.4%) in light smokers, 0.09 mmol/l (10.3%) in moderate smokers and 0.12 mmol/l (13.8%) in heavy smokers. Cigarette smoking was related to fasting triglyceride (directly) but this was removed by further adjustment for HDL-cholesterol. Cigarette smoking was not related to low density lipoprotein cholesterol. These results are compared to those of other surveys. PMID- 8225738 TI - Physical activity at work and cardiovascular disease risk: results from the MONICA Augsburg study. AB - In the MONICA Augsburg study the relationship between physical activity at work, cardiovascular risk factors, incidence of fatal and non-fatal myocardial infarction (MI) and total mortality was investigated in employed men. The MONICA Augsburg baseline survey of 1984/85 was designed as a cross-sectional study with follow-up. Physical activity was determined in 45-64 year old men by interview (n = 1074, 83.6%) and by a 7-day activity diary (n = 797, 62.0%). Employed men were categorized into an active versus inactive group based on interview data on physical activity at work (PAW) which had been validated against the 7-day activity diary. When stratified by PAW, age-adjusted means of diastolic (DBP) and systolic blood pressure (SBP) and total-cholesterol (T-C) were lower and the age adjusted mean of HDL-cholesterol (HDL-C) was higher in active than in inactive men. A linear regression model controlling for age, body mass index, alcohol consumption, smoking, and heart rate confirmed this relationship for physical activity at work; regression coefficients: DBP: beta = -2.38 (95% confidence interval [CI] : -3.85(-)-0.91); SBP: beta = -2.87 (95% CI : -5.22(-)-0.52); T-C: beta = -2.80 (95% CI : -9.31-3.71); HDL-C: beta = 3.48 (95% CI : 1.28-5.79). Survey participants were followed-up for a period of 5.8 years. The incidence density (per 1000 person-years) of fatal plus non-fatal MI was 6.8 for active men versus 5.9 for inactive men; the incidence density for total mortality was 11.2 versus 5.9, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225739 TI - Low prevalence of risk factors for coronary heart disease in rural Tanzania. AB - A community-based survey was used to assess the prevalence of risk factors for coronary heart disease (CHD) in rural Tanzanians. In all, 8581 subjects (3705 men, 4876 women) aged > or = 15 years in eight villages in three regions in rural Tanzania representing a range of socioeconomic deprivation were studied. The main outcome measures were serum cholesterol and triglyceride level, blood pressure and prevalence of dyslipidaemia, hypertension, smoking, overweight, impaired glucose tolerance (IGT) and diabetes; as well as ECG changes. Mean serum cholesterol levels in men were 4.2, 3.4 and 3.7 mmol/l, and in women 4.4 3.6 and 3.9 mmol/l in Kilimanjaro, Morogoro and Mara regions respectively. In Kilimanjaro region 17.4% of men and 19.0% of women had values above 5.2 mmol/l compared with only 5.0% and 6.7% in Morogoro region and 4.8% and 6.9% respectively in Mara region. Systolic and diastolic blood pressures increased with age in both men and women in all three regions with the most marked increase in Kilimanjaro region and the smallest rise in Mara region. Mean age-adjusted values were highest in Kilimanjaro region (124/75 mm Hg, and 125/76 mm Hg in men and women respectively) and lowest in Mara region (120/70 mm Hg in men and 118/68 mm Hg in women). Hypertension was found in 6.6% of men and 7.5% of women in Kilimanjaro region, 3.3% and 4.7% in Morogoro, and 2.6% and 3.4% in Mara region. Cigarette smoking was found in 42.6% of men in Kilimanjaro region, 28.2% of Morogoro region and 8.6% in Mara region. Less than 4% of women smoked in all three regions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225740 TI - A Thai monk: an agent for smoking reduction in a rural population. AB - Monks in a rural community in northern Thailand were observed to conduct anti smoking activities. To evaluate the effects of the monks' efforts on change of people's smoking behaviour and attitudes, we conducted a survey of adults > or = 15 years of age in this village (intervention village sample) and a randomly selected village (control village sample) in the same subdistrict. Between March and April 1991, we interviewed 372 individuals in the intervention village and 664 in the control village. The two villages were of similar socioeconomic status. The proportion of ever smokers who had tried to quit was greater in the intervention sample (79.6%) than in the control sample (72.0%) (P = 0.048). The proportion of former smokers who had stopped smoking for > or = 1 year was significantly greater in the intervention sample (25.5% of ever smokers) than in the control sample (16.6%) (P = 0.011). Of current smokers in the intervention village, 45.8% reported smoking less tobacco during the year preceding the survey than in the previous 1-year period compared with 35.2% in the control village (P = 0.034). Many former smokers (80.3%) in the intervention village cited the suggestion of a monk as one important reason for quitting compared with 25.6% in the control village (P = 0.000). The proportion of individuals who were well aware of the harmful effects of smoking on health was greater in the intervention village than in the control village. Religious leaders may be helpful in a community-based smoking prevention programme. PMID- 8225741 TI - Smoking cessation and changes in respiratory symptoms in two populations followed for 13 years. AB - To investigate the relationship between persistence and incidence rates of respiratory symptoms, and the cessation of cigarette smoking, the data from longitudinal studies conducted in Cracow, Poland and Tucson, USA were analysed jointly. Among 1722 subjects smoking at the beginning of the study, 468 had given up smoking at the 13-year follow-up. The persistence and incidence rates of chronic cough, chronic phlegm, wheeze and attacks of breathlessness were reduced by 50% in ex-smokers compared to the subjects continuing to smoke. The beneficial effects of smoking cessation were decreased in subjects smoking more cigarettes per day in the past and starting to smoke at a younger age. The symptoms were less likely if smoking ceased before the onset of any respiratory disease. These results were similar in the Cracow and Tucson populations, confirming the universal nature of the observations. PMID- 8225742 TI - Mortality in a cohort of pesticide applicators in an urban setting. AB - The mortality of a cohort of 168 pesticide applicators employed in the disinfestation service of the city of Rome for an average of 20 years is examined. Exposure data were abstracted from work records, and causes of death for the 42 workers who had died, were obtained from death certificates. Standardized mortality ratios (SMR) for specific causes of death were computed on the basis of provincial mortality rates. An excess in mortality from cancer of the liver and bile ducts with four cases observed and 0.7 expected (SMR = 571, 95% confidence interval (CI): 154-1463) was found. Increased risk for other cancers was also observed, but the SMR were not statistically different from unity. An increased risk of liver cancer occurs in those exposed to organochlorine pesticides between 1960 and 1965. PMID- 8225743 TI - Maternal recall of symptoms associated with childhood deaths in rural east Africa. AB - Verbal autopsies (VA) are frequently used to determine causes of death for individuals for whom there is no reliable clinical information regarding the terminal illness. VA interviews are used to note key symptoms and signs recalled by relatives of the deceased and diagnoses ascribed according to the symptom complexes. The VA technique assumes that individual disease entities have discrete symptom complexes and that these can be accurately recognized and recalled by the interviewees. We have examined the accuracy with which specific symptoms are recalled over time by mothers or normal guardians of 491 children who died on the paediatric wards of two district hospitals in East Africa. Kwashiorkor, measles, trauma, generalized convulsions and neonatal tetanus were all reported with a high degree of accuracy for children who died of these conditions and had low false positive rates for children without these conditions. Recall was similar within 1 month of death compared to recall after 6 months for most symptoms and signs except neonatal tetanus where false positive reports by mothers increased with time since death. Symptoms and signs commonly used to describe malaria, respiratory tract and diarrhoea-related deaths were reported by mothers to have been present during the terminal illness in 43% of cases where these features were absent. Recall abilities differed between the two communities studied for some symptoms and signs highlighting the importance of such studies in every setting where VA are applied. PMID- 8225744 TI - Breastfeeding and other reproductive factors and the risk of hip fractures in elderly women. AB - The objective of this population-based case-control study was to clarify the relationships between parity, breastfeeding and ages at menarche and menopause and the risk of hip fracture among elderly women. The study base comprised women aged 65 years and over living in a defined region in Sydney, Australia, during 1990-1991. Cases (n = 174) were recruited from 12 hospitals and controls (n = 137) were selected using an area probability sampling method, with additional sampling from nursing homes. The age- and proxy-adjusted odds ratio (OR) comparing parous women who had never breastfed with nulliparous women was 2.11 (95% confidence interval (CI): 0.85-5.25). Among parous women, the age- and proxy adjusted OR for ever versus never breastfeeding was 0.47 (95% CI: 0.22-0.99). There was a dose-response relationship between average duration of breastfeeding per child and risk of hip fracture (test for trend: P < 0.01). Age at menopause and age at menarche were only weakly associated with hip fracture risk. This study suggests that breastfeeding may protect parous women against hip fracture in old age. PMID- 8225745 TI - Validity of contraceptive histories in a rural community in Kenya. AB - To determine the validity of self-reported contraceptive histories obtained from rural Kenyan women, we interviewed 122 women participants in a long-term study during which their contraceptive use had been recorded regularly at clinics. Interview information was compared with each woman's clinic record. In all 71 women (58%) reported ever use of oral contraceptives, while 76 (62%) of the clinic records documented ever use. The agreement between the records and interview was reasonable (Kappa = 0.54, P < 0.001). The sensitivity of self report of use compared to clinic records was 79%, specificity was 76%, positive and negative predictive values were 85% and 69% respectively. Ever use of injectable contraceptives (Depo-Provera) was reported by 78 (63%) women while the records showed use by 80 (65%) women (Kappa = 0.87, P < 0.001). The sensitivity, specificity, positive and negative predictive values were 94%, 93%, 96%, and 89% respectively. For the intrauterine contraceptive device, ever use was reported by 18 (14.8%) women while the records showed 19 (15.6%) (Kappa = 0.90, P < 0.001). The associated sensitivity, specificity, positive and negative predictive values were 90%, 99%, 94% and 98% respectively. The validity of reported duration of use of each method showed a similar pattern to that of the history of ever use. The Spearman's rank correlations for the duration of use calculated from interview compared with the clinic records were for oral contraceptives r = 0.68 (P < 0.001), injectables r = 0.81 (P < 0.001) and intrauterine devices r = 0.82 (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225746 TI - Water and non-water-related risk factors for gastroenteritis among bathers exposed to sewage-contaminated marine waters. AB - All previously published epidemiological studies of the health effects of bathing in marine waters contaminated with domestic sewage contain three major methodological weaknesses in study design: (1) failure to control for the substantial amount of temporal and spatial variation in indicator organism densities shown to occur within just a few hours at marine water bathing locations; (2) failure to relate indicator organism density directly to the individual bather; and (3) failure to rigorously control for non-water-related risk factors on previously reported associations between bathing in marine waters and illness among such bathers. We report the results of two intervention follow up studies specifically designed to address these methodological weaknesses. We restricted study outcome to bathing-associated gastroenteritis since this is the illness most consistently reported to be associated with bathing in marine waters, and upon which both current US Marine Water Quality Criteria and other standards used worldwide are based. Our results show that faecal streptococci was the only indicator organism to predict the occurrence of gastroenteritis among bathers, and this occurred at only one of the three water quality sampling depths used in our study. The consumption of three different foods known or suspected to act as vectors in the transmission of gastroenteritis, as well as one non-food, non-water-related risk factor for gastroenteritis were found to significantly increase the risk of gastroenteritis among bathers. Multiple logistic regression modelling showed that these non-water-related risk factors confounded the relationship between exposure to marine waters of varying faecal streptococci densities and the occurrence of gastroenteritis among bathers to a moderate degree. Moreover, these analyses showed that the risk of gastroenteritis to the individual bather caused by these non-water-related risk factors, approximated the risk of gastroenteritis among bathers exposed to waters containing relatively high faecal streptococci densities. The implications of these findings with regard to the validity of present marine water quality criteria and on the need for, and design of, future epidemiological studies of bathing water associated illness are discussed. PMID- 8225747 TI - Clustering of xerophthalmia within households and villages. AB - The clustering of xerophthalmia within households and villages was estimated among preschool age children using data from studies conducted in Malawi, Zambia, Indonesia and Nepal over the past decade. Pairwise odds ratios (OR) were used to measure the magnitude of clustering. This OR measures the risk of xerophthalmia for a preschool child given that another randomly chosen preschool child from the same household (or same village but different household) had xerophthalmia, relative to the risk if that randomly chosen preschool child did not have xerophthalmia. Village pairwise OR ranged from 1.2 in Malawi to 2.2 in Nepal. Household pairwise OR ranged from 4.4 in Malawi to 9.7 in Indonesia, indicating that xerophthalmia clustering was much greater within households than villages. The magnitude of this clustering was as large, or larger than, infectious outcomes such as diarrhoea, fever and cough. Although xerophthalmia was associated with a weekly history of infectious morbidity, the clustering of diarrhoea, fever and cough explained very little of the xerophthalmia clustering observed in each of these studies, Hence, other household factors such as food availability and dietary practices should be examined for their role in the clustering of xerophthalmia within certain households. PMID- 8225748 TI - HTLV-I/II infections in Spain. The HTLV-I/II Spanish Study Group. AB - Antibodies to HTLV-I/II were investigated in sera from 7521 individuals living in Spain. They were classified in four major groups: a) subjects at high risk of retroviral infections e.g. parenteral drug addicts, homosexuals, prostitutes, and multiple-transfused individuals; b) patients suffering illness associated with HTLV-I in endemic regions; c) immigrants from endemic areas; and d) blood donors. Sera were collected from 1984 to December 1991. Repeatedly reactive ELISA was found in 211 samples (2.8%), but Western blot only confirmed the presence of HTLV I/II antibodies in 23 samples (0.30%), corresponding to eight (0.25%) out of 3207 drug abusers, six (0.72%) out of 894 immigrants (five Africans and one South American), three (0.41%) out of 727 patients with HTLV-related diseases (one woman with HTLV-I associated myelopathy had received blood transfusions in an endemic area), four (0.54%) out of 793 prostitutes, one multiple-transfused native woman, and one (0.16%) out of 603 native seamen. The Western blot antibody pattern confirmed HTLV-II infection instead of HTLV-I in nine (39%) subjects. The remaining 14 (61%) HTLV-reactive samples were interpreted as HTLV-I seropositive, most of which were from immigrants. None of 857 blood donors analysed was reactive for HTLV antibody. These results suggest that both HTLV-I and HTLV-II are present in Spain, although at a low rate and mostly restricted to individuals coming from endemic areas, drug addicts, and prostitutes. Furthermore, diseases related to HTLV-I (particularly lymphoproliferative disorders, and subacute myelopathies) seem to be rarely associated with these viruses in Spain, a non endemic area. PMID- 8225749 TI - The design and analysis of cholera vaccine trials: recent lessons from Bangladesh. AB - The recent spread of cholera to Latin America, together with the persistent burden of this disease in Asia and Africa, have stimulated efforts to evaluate new cholera vaccines in field settings. Although the standard experimental paradigm for vaccine field trials is well established, the success of these trials will also depend on suitable consideration of the epidemiology of cholera and of cholera vaccination in the setting under study. Epidemiological studies done in Bangladesh emphasize the importance of appreciating the poorly predictable, multifocal occurrence of cholera in estimating a probable incidence of cholera for a field trial. They also underscore how the filtering effect of enrolling subjects into a prospective trial can dramatically reduce the available population for study, and can yield a study sample whose expected risk of cholera differs markedly from that for the source population. Finally, the data highlight the subtle effects that the mode of surveillance and the choice of an outcome definition can have upon protective efficacy, and emphasize the need for subgroup analyses that address the distinctive variations in vaccine protection that may occur in subjects differing in age and in ABO blood groups, and in subjects exposed to classical versus El Tor cholera. PMID- 8225750 TI - The dot map as an epidemiological tool: a case study of Schistosoma mansoni infection in an urban setting. AB - In this paper dot maps are used as an epidemiological tool to describe the spatial patterns of cases, non-cases and some relevant risk factors associated with the occurrence of Schistosoma mansoni infection in an urban environment. The study was carried out in Santo Antonio de Jesus, a town in north-east Brazil with a population of around 45,000 inhabitants. The survey was targeted at all children aged between 12 years 8 months and 14 years 11 months who were living in the town at the time of the survey. An extensive questionnaire was used to collect information on each child and on family and household conditions; stool samples were also taken for quantitative examination. A computerized digitizer was used on a map (scale 1:4000) to create x and y coordinates for each child's household. The result was matched with individual data from the questionnaires and the stool examination. From this data base it was possible, by using a statistical package, to make epidemiological analyses and by using the statistical package associated with a geocartographic system package it was possible to dot plot selected values of defined variables. These techniques substantiated the findings on the risk factors at a local level and, as a consequence, improve our ability to define targeted actions towards local control of schistosomiasis. PMID- 8225751 TI - Estimation of vaccine effectiveness using the screening method. AB - The screening method provides a simple and rapid way of estimating vaccine effectiveness. The paper discusses the validity of the screening method with particular reference to bias and precision. Methods for correcting confounding, adjusting for covariates and over-dispersion, and deriving confidence limits are discussed in a modelling framework. The methods are illustrated using data on measles and pertussis vaccines. PMID- 8225752 TI - Validation of self-report of literacy among periurban women in Bolivia. AB - Many surveys ask women to rate their literacy level, but the validity of these self-reports is not usually evaluated. In this study of infant health, conducted in Cochabamba, Bolivia, maternal literacy was assessed by the mother herself and by the interviewer, based on the mother's reading a short text aloud. The two ratings were highly correlated (P < 0.001). This finding suggests that maternal self-reports of literacy are very reliable. PMID- 8225753 TI - Infant mortality and abortion in HIV-1 infected women. PMID- 8225754 TI - Maternal smoking during pregnancy and delinquency of the offspring: an association without causation? PMID- 8225755 TI - Why do some parasitic nematodes secrete acetylcholinesterase (AChE)? PMID- 8225756 TI - Mucosal mast cells and the expulsive mechanisms of mice against Strongyloides venezuelensis. AB - The possible importance of mucosal mast cells in the expulsive mechanisms of mice against Strongyloides venezuelensis was examined. After a primary infection by subcutaneous inoculation with various doses into C57BL/6 mice, about 50% of the initial dose of infective larvae (L3) became adult worms and, regardless of the dose of infection, they were completely expelled by Day 12 with similar kinetics. Intestinal mastocytosis at the time of expulsion was comparable among groups given different doses of infection. A kinetic study after infection with 2000 L3 in C57BL/6 mice revealed that mastocytosis started from Day 8, rapidly reached a peak on Day 12, and then gradually decreased. The strongest mastocytosis was observed in the upper one sixth of the small intestine where the majority of adult worms parasitized. Over 80% of mast cells induced by the infection were located in the intestinal epithelial layer. When mast cell-deficient W/Wv and their normal littermate +/+ mice were infected with 1000 L3, expulsion was significantly delayed in W/Wv mice, though adult worms were eventually expelled by Day 18 in W/Wv mice. Delayed expulsion as well as defective mast cell responses of W/Wv mice were completely restored by bone marrow grafting 10 weeks prior to infection. These results show that, like S. ratti infection, intestinal mucosal mast cells are important in causing expulsion of S. venezuelensis. PMID- 8225757 TI - Metabolic disturbances in Plasmodium coatneyi-infected rhesus monkeys. AB - To investigate metabolic disturbances in an animal model of human malaria, four rhesus monkeys (Macaca mulatta) were infected with Plasmodium coatneyi, a parasite which induces cytoadherence of infected erythrocytes. When moribund or the parasitaemia had plateaued, the monkeys were sacrificed (3 animals) or treated with chloroquine (1 animal). Blood and cerebrospinal fluid (CSF) were sampled at intervals between inoculation and sacrifice or treatment. Arterial and CSF glucose and lactate rose during infection, indicating evolving insulin resistance. The arteriovenous difference in glucose concentration also increased, consistent with increased glucose consumption by parasitised tissues. Arterial plasma lactate rose but a positive arteriovenous concentration difference suggested tissue lactate uptake. The animal with the highest plasma lactate at sacrifice remained hyperglycaemic but also had the highest CSF lactate, the greatest cerebral sequestration and neurological depression, and biochemical and histological evidence of severe hepatic pathology. Serum cholesterol and corrected serum calcium fell consistently during infection; serum phosphate was also reduced in animals without renal impairment. These preliminary results indicate that lactic acidosis is a late complication of severe malaria and, by implication from this and other studies, hypoglycaemia occurs even later; other metabolic changes during P. coatneyi infection in rhesus monkeys also parallel those of severe falciparum malaria in humans. The model could be used in further studies of malaria-associated metabolic dysfunction and its management. PMID- 8225758 TI - High performance latex reagent for hydatid serology using an Echinococcus granulosus lipoprotein antigen fraction purified from cyst fluid in one step. AB - A lipoprotein fraction from fertile bovine hydatid cyst fluid (FBHCF) was isolated by affinity chromatography on acrylic-heparin. The heparin-binding lipoprotein fraction (HBLF) was proved to be free from host immunoglobulins and albumin. Immunoblotting with confirmed human hydatid sera showed that the major relevant diagnostic bands in FBHCF were also present in HBLF. HBLF or FBHCF were used to sensitize polystyrene latex particles. HBLF-latex showed both higher reactivity with positive hydatid human sera and higher batch-to-batch reproducibility than FBHCF-latex. Titration of sera from 119 surgically confirmed hydatid patients, 48 with other parasitic diseases, 6 with unrelated diseases and 37 healthy subjects was performed using HBLF-latex and conventional ELISA (using FBHCF). Sensitivity and specificity were 83 and 87% for ELISA and 87 and 88% for HBLF-latex. These results indicate that, while being simpler and faster to perform, the HBLF-latex has a similar diagnostic value to that of the conventional ELISA test. PMID- 8225759 TI - An in vitro assay utilising parasitic larval Haemonchus contortus to detect resistance to closantel and other anthelmintics. AB - A new assay for detecting resistance to anthelmintics in vitro is described. The assay uses a simple culture system in which the ability of anthelmintics to kill or inhibit the migration of parasitic third and fourth stage Haemonchus contortus larvae through a 50 microns aperture mesh is assessed. The assay detects 2-10 fold resistance to closantel. Resistance to benzimidazoles, levamisole and ivermectin can also be detected. PMID- 8225760 TI - Serum antibodies in canine echinococcosis. AB - Specific IgG, IgA and IgE antibodies against E. granulosus protoscolex antigen were detected by enzyme-linked immunosorbent assay (ELISA) in sera from dogs experimentally or naturally infected with E. granulosus. The specificities of the IgG, IgA and IgE ELISAs were 100, 100 and 97.3%, respectively. Sera from 626 dogs of different categories and geographic regions in Australia, Uruguay and Kenya were tested. There were distinct differences in antibody responses in experimentally infected canids and in the number of naturally infected dogs found seropositive, depending on geographic region. The overall sensitivities of the ELISA (IgG, IgA and IgE) ranged between 73 and 84%, except for one geographic region where it was 54%. Genetic differences of the dogs and/or antigenic variations of the parasite appear to be responsible for the variations in specific antibody levels in infected dogs. In average, approximately one third of dogs from hyperendemic hydatid regions, without E. granulosus worms at autopsy or negative for E. granulosus infection by arecoline testing, were seropositive for anti-E. granulosus antibodies, suggesting previous infection with or exposure to the parasite. The results of this study demonstrate that, although the diagnosis of current intestinal E. granulosus infection on an individual dog basis is not always reliable by serology, serum antibody ELISA is useful as an epidemiological/educational tool for seroprevalence studies on canine echinococcosis. PMID- 8225761 TI - Dynamics of rat muscle mitochondria uncoupling in trichinellosis. AB - Parallel oxygraphic measurements of respiratory control index (RCI) and spectroscopic measurements of the activity of two mitochondrial enzymes from rat muscle, mtATPase and succinate dehydrogenase were performed. In Trichinella spiralis infection, in comparison with Trichinella pseudospiralis infection, a delayed stimulation of mtATPase in 3rd week p.i. was observed and the degree of stimulation was weaker. A second peak of mtATPase stimulation appeared at the beginning of the encystation phase. The two-phase phenomenon was accompanied by a sharp decrease of RCI to 39% of normal values and almost 2-fold increase in the ratio of Mg(2+)-stimulated to Mg(2+)-unstimulated mtATPase. By the end of investigations (7th week) the level of SDH normalized, but mtATPase was still elevated. The results, as in the case of T. pseudospiralis infection, support the testing of mtATPase activity to follow bioenergetic changes in tissue parasite infections. In comparison with oxygraphic measurements ATPase testing needs a much smaller amount of biological material and ATP. PMID- 8225762 TI - Giardia intestinalis: detection of major genotypes by restriction analysis of gene amplification products. AB - The polymerase chain reaction (PCR) has been used to amplify a 0.52 kb segment of Giardia intestinalis DNA, using primers specific for nucleotide sequences conserved within two genes (tsp11 and tsa417) that encode homologous, cysteine rich trophozoite surface proteins. Using products amplified from axenic isolates belonging to genetic groups I and II (defined on the basis of allozyme electrophoresis data), restriction endonuclease analysis revealed both tsp11-like and tsa417-like fragments within all samples. The study also identified among the amplification products of group II organisms an additional fragment, containing a novel PstI site, that is not detected in the reaction products of group I isolates. The recovery of three distinct PCR products from each group II isolate was verified by cloning the fragments into the plasmid vector pGEM-7. Fragments containing the new PstI site possess the ClaI site common to both tsp11 and tsa417-like fragments, but they lack the HindIII site which characterizes tsp11 like fragments and also lack the PstI and KpnI sites which characterize tsa417 like fragments. Spot-blot analyses using cloned fragments of all three types as probes showed strong homologous hybridization but weak heterologous hybridization, indicating that each type differs substantially in nucleotide sequence from the others. Because the samples of Giardia DNA used in the PCR were purified from cultures that had been established from single trophozoites, the data indicate that individual trophozoites belonging to genetic group II possess three homologous genes defined by these related fragments. The presence of a PstI site in the amplified segment of the newly-discovered third gene of group II organisms provides a simple diagnostic means of differentiating group I and II isolates. PMID- 8225763 TI - Chromosomal duplication in Giardia duodenalis. AB - Strains of Giardia duodenalis from diverse parts of the world have three or four chromosomal bands in the range of 650-800 kb as defined by field inversion gel electrophoresis. The extra chromosome band in this range defined a group of strains which are geographically distinct from other strains missing this band. The cloned line WB-1B has three chromosome bands in this size range and chromosome band 3 was used to construct a library of chromosome-specific probes. In some strains examined, including BRIS/83/HEPU/106, a subset of these WB-1B probes hybridized to chromosome band 3 and to the extra chromosome band 4, indicating a partial duplication of chromosome 3 in BRIS/83/HEPU/106. This duplication was estimated to be at least 500 kb when the sizes of NotI chromosome segments which hybridized with chromosome band 3-specific probes were added. A second subset of WB-1B chromosome 3-specific probes hybridized to a fifth chromosome of strain BRIS/83/HEPU/106, in the size range 650-800 kb, which was not visible by ethidium bromide staining. The two subsets of WB-1B probes hybridized to a variety of chromosome combinations in this size range in other Giardia strains and included apparent reversal of chromosome 3 and 4 mobility as well as identification of other minor chromosomes. These data indicate that chromosome band 3 of the line WB-1B is a cluster of at least two different chromosomes that cannot be electrophoretically separated but genetic rearrangements in other strains allow separation of linkage groups carried by chromosome band 3 of WB-1B.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225764 TI - A cytochemical study of the nervous system of the proteocephalidean cestode, Proteocephalus pollanicola. AB - The localization and distribution of cholinergic, serotoninergic and peptidergic nerve elements in the proteocephalidean tapeworm, Proteocephalus pollanicola, have been investigated by enzyme histochemistry, and by an indirect immunofluorescence technique interfaced with confocal scanning laser microscopy. Cholinesterase (ChE) activity was localized in the major components of the central nervous system (CNS) and the peripheral nervous system (PNS), including the innervation of the reproductive structures of the worm. Serotoninergic (5-HT) nerves were found in the paired cerebral ganglia, transverse commissure and in the 10 longitudinal nerve cords. Antisera to 17 mammalian regulatory peptides and the invertebrate peptide FMRFamide have been used to explore the peptidergic nervous system of the worm. The most extensive immunostaining occurred with antisera raised to members of the neuropeptide Y superfamily, namely neuropeptide Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP). In all cases, intense immunoreactivity was found in numerous cell bodies and fibres of both the CNS and PNS, including the innervation of the reproductive apparatus. FMRFamide antisera stained the same structures to a comparable degree as those raised to the NPY superfamily. Cholinergic and peptidergic elements were much more prevalent within the CNS, while the serotoninergic nerve fibres tended to dominate in the PNS. The overlap obtained in staining patterns for the peptidergic and cholinergic components suggests that there may be a certain amount of co-localization of peptides with small-molecule transmitter substances in the same neurone. Weak staining for the tachykinin, substance P and for calcitonin gene-related peptide (CGRP) was confined to the major longitudinal nerve cords. PMID- 8225765 TI - The effect of experimental trichostrongyle infections of housed young calves on the subsequent course of natural infection on pasture. AB - The present studies were designed to investigate whether experimental, mixed trichostrongyle infections of stabled calves prior to their first grazing season could confer sufficient immunity to significantly reduce egg excretion after turnout, and thereby prevent loss-producing infections later on. The study comprised four groups, each of seven calves. During spring, two of the groups received two different dose levels of infective larvae twice weekly, and one group received larger larval doses at monthly intervals. One group served as non experimentally exposed controls. In May all groups grazed separately on similarly contaminated plots. All experimental groups had reduced egg counts, and herbage infectivity of their plots was significantly lower than that of the controls. These findings were reflected in serum pepsinogen levels and in specific serum antibody responses. Additionally, clinical disease was only observed in the control group animals. PMID- 8225766 TI - In vitro interleukin-1 and tumor necrosis factor-alpha production by macrophages from chickens infected with either Eimeria maxima or Eimeria tenella. AB - We measured the in vitro production of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF- ) by macrophages from chickens during and immediately following an infection with one of two different species of Eimeria, E. maxima and E. tenella. Quantitatively, the amounts of IL-1 produced during each infection were nearly identical regardless of the oocyst dose of each parasite. TNF production followed a biphasic pattern of increased production with the first peak associated with the pathogenesis of disease and the second peak associated with the development of protective immunity. These experiments together with others we have reported imply cells taken from chickens infected with coccidia have a greater capacity to produce cytokines upon stimulation in vitro than cells from non-infected birds. The production of significantly greater amounts of TNF during the days 3-6 after inoculation correlates with the appearance of the most characteristic local and systemic pathophysiological changes in the host induced by the coccidia. The excessive release of TNF in response to a heavy coccidial infection may account for many of the pathological features observed with avian coccidiosis probably through the release of other mediators. PMID- 8225767 TI - A case of spirurid larva infection in the anterior chamber of the human eye. AB - A 39-year-old male living in Yamanashi Prefecture, Japan, who complained of headaches, and redness, the sensation of foreign bodies and puffy eyelids of the left eye, visited the Department of Ophthalmology, Yamanashi Medical College. A larval nematode was found in the anterior chamber of the left eye and removed through a limbic incision. This nematode was identified as the spirurid type X larva. PMID- 8225768 TI - The effect of age on the response of Romney sheep to gastrointestinal nematodes during grazing. AB - Three groups of Romney sheep, reared worm-free in pens from birth until 4, 16 or 28 months of age, were permitted to graze nematode-infected pasture for 4 weeks after which they were returned to pens and slaughtered 4 weeks later. Three groups of three Romney sheep reared and maintained worm-free for similar periods served as uninfected control groups. Faecal egg counts (FECs) 8 weeks after grazing commenced were significantly greater in 4-month-old sheep than the 28 month-old animals. At slaughter, no difference in nematode burdens among these three age groups was found. Histological examination of the abomasal and small intestine mucosa, taken from sheep at slaughter, revealed significantly greater numbers of globule leukocytes/mucosal mast cells (GL/MMC) bur fewer connective tissue type mast cells (CTMC) in sheep aged 16 and 28 months than in those aged 4 months. The numbers of eosinophils in the gastrointestinal mucosal tissues of the three groups of infected sheep were not significantly different. In uninfected sheep the numbers of eosinophils, GL/MMC and CTMC in the mucosal tissues did not differ between the three age groups and the numbers of GL/MMC and CTMC were similar to those in 4-month-old infected sheep. Eosinophil numbers in the mucosal tissue of the infected groups were significantly higher than in the uninfected groups. In the infected groups, FECs but not worm burdens were significantly negatively correlated with GL/MMC numbers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225769 TI - Human humoral immune responses against Trichinella spiralis. AB - The analysis of trichinellosis patients' sera was performed by Western blot against a crude extract of Trichinella spiralis. Eight distinctive bands from 38 to 104 kDa were present in most of the patients' sera as a striking homogeneous response. Some of these characteristic bands were found to be recognized by the serum of infected rodents and of an immunized rabbit in this work, and have also been reported to be recognized by monoclonal and polyclonal antibodies from T. spiralis infected or immunized rats, mice, swine or rabbits. Our data indicate that different animal species as well as humans recognize unique immunodominant epitopes of T. spiralis. PMID- 8225770 TI - MHC haplotype influences primary Giardia muris infections in H-2 congenic strains of mice. AB - The course of primary Giarda muris infections was studied in H-2 congenic strains of mice on the BALB and C57BL/10 (B10) genetic backgrounds. Infection was curtailed more rapidly in B10 strains, compared with BALB strains, and unaffected by H-2 haplotype. On the other hand the duration of infection in BALB strains did vary with H-2 haplotype with BALB/B(H-2b) mice taking significantly longer to clear infection than BALB/c(H-2d) and BALB/K(H-2k) mice. These experiments demonstrate that both MHC and non-MHC genes influence the outcome of primary G.muris infections. PMID- 8225771 TI - A survey of didymozoid trematodes of the barracuda Sphyraena obtusata from Kuwait Bay. AB - Examination of 120 yellow-finned barracudas, Sphyraena obtusata, from Kuwait Bay revealed 8 species of didymozoid trematodes encapsulated in pairs in the tissues. Four species of the didymozoids, Allodidymozoon operculare, Didymocylindrus sp., Didymocystis sp., and Didymocystoides sp., were found in the head region, 3 species, Didymozoon tetragynae, Lepidodidymozoon sinicum and Platocystoides polyaster, in external surfaces and 1 species, Allodidymozoon sp., in stomach wall. The didymozoid species collected constitute new geographic distribution records and occurrence of D. tetragynae and L. sinicum are new host records. PMID- 8225772 TI - Influence of bacteria on electrophoretic proteinase patterns of Entamoeba histolytica isolates. AB - Gelatin SDS- polyacrylamide gel electrophoresis was used to compare proteinase banding patterns under reducing conditions from whole cell lysates of four axenic and four xenic pathogenic strains of Entamoeba histolytica. All strains shared major bands in the 34 and 66-68 kDa regions, whereas only the axenic strains produced major bands at 26, 28, 30 and 45 kDa. One axenic strain, NIH 200, when reassociated with mixed bacterial flora, reverted to an electrophoretic banding pattern characteristic of other xenic strains. These results suggest that the 26 30 kDa and 45 kDa proteinases of E. histolytica are induced by bacterial starvation while others are constitutively expressed. It is also proposed that the axenic bands of 45 and 34 kDa represent respectively, the reduced forms of the 56 and 40 kDa bands reported elsewhere under non-reducing conditions. PMID- 8225773 TI - Killing of Trichomonas vaginalis by complement-mediated lysis is not associated with the presence of Trichomonas vaginalis virus. AB - We have examined the possible relationship between trichomonad killing by human serum and the presence of virus-encoded double-stranded ribonucleic acid in T. vaginalis (TVV). Indirect immunofluorescence assay revealed that non-immune serum (NIS) and T. vaginalis-immune serum (TVIS) had immunoglobulin G (IgG) antibody titres of 1:8 and 1:256, respectively, against T. vaginalis. Among the 12 isolates of T. vaginalis examined, 9 were infected with TVV. Upon long-term (> 9 months) culture, of the 9 infected isolates, 3 isolates lost the virus during the passage process. Five of 9 TVV-infected isolates were completely killed by 10% NIS while the other 4 TVV-infected isolates had viability over the range 22-81%. Three fresh non-TVV-infected isolates had viability over the range 12-89%. On the other hand, no trichomonads survived in the presence of 10% TVIS. Viability of the virus-lost isolates during long-term culture was not altered when compared with that of their corresponding fresh isolates. Heat-inactivated-NIS and -TVIS had no killing effect on trichomonads while absorbed-NIS and -TVIS (ATVIS) had a similar killing effect to NIS. Further studies on the role of antibody and complement in the killing of trichomonads by serum revealed no significant difference in trichomonad viability between treatments of Mg2+-ethylene glycol bis-(beta-aminoethyl ether)- N,N,N1,N1- tetraacetic acid (Mg(2+)-EGTA)-TVIS and of Mg(2+)-EGTA-ATVIS. Zymosan-treated ATVIS did not kill trichomonads but zymosan treated TVIS had a marked killing effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225774 TI - Morphology of Blastocystis sp. isolated from circus animals. AB - Blastocystis sp. is reported for the first time from faecal samples collected from a camel, a llama, a highland bull and a lion in a travelling circus. Fresh faecal specimens were examined by light and electron microscopy, and vacuolar and cyst forms of similar morphology were present in all three ungulates. These cells were smaller than cultured vacuolar cells of Blastocystis hominis isolated from humans and contained only a single vacuole in comparison to the multivacuolar cell found in fresh human faeces. The taxonomic relationship of Blastocystis isolated from humans and ungulates remains to be determined. The number of parasites present in the lion sample was too small to make valid comparisons. PMID- 8225775 TI - Immunity in vaccinated cattle exposed to experimental and natural infestations with Boophilus microplus. AB - Gut membranes of the cattle tick Boophilus microplus were extracted using a low ionic strength buffer and the soluble antigens were used to vaccinate Hereford cattle. Following either experimental infestation with 40,000 larval ticks or exposure to field infestation, ticks from vaccinated cattle laid 54 and 75% fewer eggs, respectively, than ticks from control cattle. After cattle were given booster injections of vaccine, and either experimentally infested with 20,000 larval ticks or exposed to field infestation, the weight of eggs laid by ticks from vaccinated cattle was reduced by 80% and 52%, respectively. Acquired immunity to the ticks varied considerably in both vaccinated and non-vaccinated cattle. PMID- 8225776 TI - Quantitative structure-activity relationship (QSAR) study of elastase substrates and inhibitors. AB - One hundred Suc-X-Y-Ala-pNA peptides: (Suc: succinyl, pNA: p-nitroanilide, X, Y: Gly, Ala, Val, Leu, Ile, Phe, Pro, alpha-aminobutyric acid, norvaline, norleucine) were synthesized and their reaction constants with porcine pancreatic elastase (Km, kcat and kcat/Km) were determined. These reaction constants were quantitatively analyzed using the Free-Wilson/Fujita-Ban method. The contribution of amino acid side chains to the reaction constants Km, kcat and kcat/Km, expressed logarithmically, was found to be additive. On the other hand, 19 elastase inhibitors of the general formula CF3CO-X-Y-Ala-pNA (X,Y: ten amino acids) were synthesized, and their inhibition constants were compared with the Michaelis constant for the corresponding substrates and analyzed using free energy-related substituent constants. In the analysis of amino acid side chains in the Y position, the Ki value of the inhibitor was generally correlated to the Km value of the substrate, which corresponded to the inhibitor, thus confirming the validity of the equation. log(1/Ki) = 1.271 log(1/Km) + 4.831 This study may serve as a prototypical approach to unraveling structure-activity relationships of peptide substrates and inhibitors of medicinal or agricultural importance. PMID- 8225777 TI - Conversion of proendothelin-1 into endothelin-1 by aspartylproteases. AB - The ability of cathepsin D, chymosin, pepsin and renin to produce endothelin-1 (ET-1) from proendothelin-1 (proET-1) was compared. No significant conversion was observed when proET-1 was incubated with up to 1 U of renin for 15 min at 37 degrees C. Cathepsin D generated, as well as degraded, ET-1 rapidly. Net production of ET-1 reached a maximum when 0.003 U of cathepsin D was used, and about 16% of the initial proET-1 was detected as ET-1 by HPLC. Pepsin up to 1 U converted proET-1 into ET-1 dose-dependently with a maximum of 71% conversion. A further increase of the amount of pepsin in the reaction mixture produced nonspecific cleavage of ET-1. Less than 10% of ET-1 remained in the presence of 15 U of pepsin. Chymosin also generated ET-1 dose-dependently, and a complete conversion was obtained at 1 U of enzyme. Greater than 1 U of chymosin only slightly degraded ET-1; at least 80% of ET-1 was still present when 15 U of chymosin was included in the assay. Other properties associated with the conversion of proET-1 into ET-1 by chymosin were investigated. Similar to authentic ET-1, the product of chymosin treatment caused contraction of isolated rabbit aortic rings, and pre-incubation of chymosin with pepstatin A abolished this contractile response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225778 TI - New C-protective group for peptide synthesis. Application of 2-(1-adamantyl) propanol-2 esters. AB - To investigate the possibility of using the 2-(1-adamantyl)-2-propyl (Adp) ester group for carboxyl protection during peptide synthesis, the tetrapeptide Boc-Phe Arg(NO2)-Phe-Pro-OAdp (IV) was prepared by the carbodiimide method. In this synthesis the Z group was removed by transfer hydrogenation and the Nps group by treatment with 2-thiopyridone. The Adp group was then cleaved with 3% TFA/DCM, yielding Boc-Phe-Arg(NO2)-Phe-Pro-OH. For subsequent kinetics studies the decapeptide Boc-Leu--Z-Orn-Arg(NO2)-Pro- Pro-Gly-Phe-Ser(Bzl)-Pro-Pro-OAdp (VIII) was synthesized by a 9 + 1 scheme. This peptide was also selectively deblocked. Comparing reaction abilities of Z-Pro-OAdp and Z-Pro-OBut, it was demonstrated that the Adp ester is cleaved by 3% TFA/DCM 230 times faster than the Bu(t) ester. For peptides IV and VIII the ratios between the rates of competitive elimination of the Adp and Boc groups by 3% TFA/DCM are 19 and 64, respectively. PMID- 8225779 TI - Inhibition of binding of phospholipase C gamma 1 SH2 domains to phosphorylated epidermal growth factor receptor by phosphorylated peptides. AB - A series of tyrosine-containing peptides 1-12: [formula: see text] (six pairs with and without the tyrosine phosphorylated) has been synthesized. The peptides were derived from tyrosine autophosphorylation sites in the epidermal growth factor receptor (EGFR): Tyr 992, 1068, 1148 and 1173. Peptide 1, derived from the Tyr 992 site, inhibited binding of a 35S-labelled fusion protein containing both of the SH2 domains from PLC gamma 1 to the phosphorylated EGFR with an IC50 of 8 microM. All of the phosphorylated peptides except 11 (1, 3, 5, 7 and 9) inhibited this binding to some degree (20-55%) at 10 microM. The nonphosphorylated peptides were inactive in this assay. The nonphosphorylated peptides 2, 4, 6, 8, 10 and 12 were obtained by standard solid-phase synthetic methodologies using both Boc/benzyl and Fmoc/tert-butyl strategies. The phosphorylated peptides 1, 3, 5, 7, 9 and 11 were similarly obtained using a Fmoc/tert-butyl strategy incorporating unprotected N alpha-Fmoc-Tyr, followed by phosphitylation and oxidation of the tyrosine in the resin-bound peptide. In addition, Asp-Ala-Asp Glu-Phe992(4-CH2PO3H2)-Leu-Ile-Pro-Gln-Gln-Gly-O H (15), an analog of 1 incorporating an enzymatically stable phosphotyrosine mimic, 4-phosphonomethyl-L phenylalanine, was synthesized and found to be inactive. PMID- 8225780 TI - Control of peptide disulfide regioisomer formation by mixed cysteine penicillamine bridges. Application to endothelin-1. AB - While incorporation of penicillamine residues (Pen; beta,beta-dimethyl cysteine) into a peptide can cause dramatic changes in biological activity, the tendency of Pen to form mixed disulfides should also allow the exploitation of the steric bulk of the beta-methyls as a synthetic device to control the production of disulfide isomers. That is, oxidation of a peptide containing an equal number of Cys and Pen residues should predominantly form products which contain mixed Cys Pen disulfides. Endothelin (ET) is a 21 amino acid peptide which contains Cys at positions 1, 3, 11 and 15. While oxidation of ET tetrathiol has been reported to produce a 3:1 ratio of the natural 1-15, 3-11 to the unnatural 1-11, 3-15 isomers, we show that oxidation of ET analogs containing two cysteines and two penicillamines predominantly formed products containing Cys-Pen disulfides. Random oxidation (air, aqueous NH4OH) of the tetrathiols of [Pen1,11, Nle7]-ET-1 or [Pen3,15, Nle7]-ET-1 produced the correct 1-15, 3-11 isomer in > 12:1 and > 22:1 ratios, respectively. Oxidation of the tetrathiol of [Pen1,15, Nle7]-ET-1 favored the unnatural 1-11, 3-15 isomer by a 4:1 ratio, indicating that a normally contrathermodynamic disulfide isomer can become the favored product as a result of the driving force for penicillamine mixed disulfide formation. Disulfide isomers were identified using ion-spray mass spectrometry in conjunction with enzymatic and acid hydrolysis. [Pen1,11, Nle7]-ET-1 was a partial agonist at the ETA receptor (EC50 = 7.5 nM in rabbit carotid artery rings; Kd = 4.5 nM in rat A10 cell membranes) while [Pen3,15, Nle7]-ET-1 (EC50 = 0.9 nM; Kd = 0.7 nM) was a full agonist with similar potency to ET-1. PMID- 8225781 TI - Small-scale manual multiple peptide synthesis system. Application to phosphopeptide synthesis. AB - A system for small-scale (ca. 10-50 mumol) manual multiple peptide synthesis assembled from commercially available solid-phase extraction apparatus is described. This system was used to prepare (on a 15 mumol scale) the five monophosphorylated isomers of the peptide ASTTVSKTE, a proteolytic fragment of the C-terminal region of rhodopsin. The peptides were assembled using serine or threonine active esters without hydroxyl protection at the positions of phosphorylation. Phosphate groups were introduced using postassembly phosphitylation/oxidation according to a published procedure [Andrews, D.M., Kitchin, J. & Seale, P.W. (1991) Int. J. Peptide Protein Res. 38, 469-475; Staerkaer, G., Jakobsen, M.H., Olsen, C.E. & Holm, A., Tetrahedron Lett. 32, 5389 5392; Perich, J.W. (1992) Int. J. Peptide Protein Res. 40 134-140]. The reported system provides a relatively inexpensive approach to multiple peptide synthesis, including synthesis of phosphopeptides, for laboratories whose synthesis requirements do not justify investment in an automated multiple peptide synthesis instrument. PMID- 8225782 TI - 3-Dimethylphosphinothioyl-2(3H)-oxazolone (MPTO), a promising new reagent for racemization-free couplings. AB - 3-Dimethylphosphinothioyl-2(3H)-oxazolone (MPTO) was synthesized, and its ability to effect racemization-free couplings and cyclization of a peptide and its C terminal epimer was examined. MPTO showed good reactivity in aprotic polar solvents such as N,N-dimethylformamide (DMF) and N-methylpyrrolidone. In reactivity MPTO resembles DPPA and dimethylphosphinothioyl azide (MPTA) previously developed by us, but it is much better than these reagents because the side reactions specific to the azide method could be avoided. In coupling of Z Gly-Val-OH with H-Val-OMe in DMF at 0 degree C, no racemization was observed without use of racemization-suppressing additives. Slight racemization observed at room temperature could be completely suppressed by addition of HOBt but not by HOSu. The utility of MPTO was demonstrated by the synthesis of cyclo-(D-Trp-D Glu(OBzl)-Ala-D-Val-Leu), an intermediate for an endothelin-binding inhibitor BE 18257A. In a comparative study using DPPA, MPTA and MPTO, no racemization was observed for MPTA or MPTO, while DPPA caused considerable racemization. When MPTO was used in the presence of HOBt rapid cyclization (3 h at RT) occurred to give the optically pure cyclic product. PMID- 8225783 TI - Preparation of activated esters of N-alkoxycarbonylamino and other acids by a modification of the mixed anhydride procedure. AB - Generation of a mixed anhydride using ethyl or isopropyl chloroformate and N methylmorpholine in dichloromethane at room temperature, followed by addition of excess ring-substituted phenol or N-substituted hydroxylamine and a catalytic amount of tertiary amine, provides an efficient synthesis of activated esters. PMID- 8225784 TI - Synthesis and in vitro activities of new tryptophan-modified and thiomethylene containing pseudopeptide antagonists of the neurokinins. AB - A series of pseudopeptide analogs of the substance P-like hexapeptide Ava-Phe-Phe Gly-Leu-Met-NH2 was produced by N alpha-protection, introduction of the thiomethylene bond, of D- and non-proteinogenic amino acids, and alteration of the side chain of tryptophan. Synthesis of the pseudopeptides on a solid phase was successfully improved by direct formation of the CH2-S bond on the resin. However, while thiomethylene formation between leucine and norleucine led to the expected SS diastereoisomer, the major product of the similar coupling between two phenylalanines was the SR isomer. An improved resistance of the analogs to proteolysis was observed, which could be related to the structural changes. Interestingly, these modifications led to three water-soluble and potent neurokinin antagonists on classical in vitro bioassays. PMID- 8225785 TI - Crystal structure and conformation of two N-tosyl-protected dipeptides containing amino acids with polar side-chains. AB - X-Ray diffraction studies and energy-minimization calculations were carried out on two dipeptides, N-tosyl-L-Ser-Gly-OMe monohydrate (C13H18N2O6S.H2O, compound A) and N-tosyl-L-Thr-Gly-OMe (C14H20N2O6, compound B). Compound A crystallized in the monoclinic system, space group P2(1) with unit cell parameters a = 4.915(1), b = 15.625(4), c = 11.003(1) A, beta = 91.28(1) degrees, V = 844.8 A3. M(r) = 348.4, d = 1.37(2) g cm-3, Z = 2, lambda(Cu K alpha) = 1.5418 A, mu = 1.99 mm-1, T = 293 K. R = 0.032 for 1451 unique reflections with I > 2 sigma(I). Compound B crystallized in the orthorhombic system, space group P2(1)2(1)2(1), with unit cell parameters a = 5.050(2), b = 16.483(3), c = 20.769(5) A, V = 1729.3 A3, Z = 4. M(r) = 344.4, d = 1.32(2) g cm-3, mu(Cu K alpha) = 1.90 mm-1. R = 0.040 for 1060 unique reflections with I > 2 sigma(I). The major difference in the backbone conformation of the two compounds is in their glycine residues, with the glycine residue in compound A adopting an extended conformation with phi = -132.6(3) degrees and psi = 175.3(3) degrees and that in compound B having a folded conformation with phi = -56.3(6) degrees and psi = -42.6(7) degrees. In compound A the oxygen atom of the Ser side-chain and the carbonyl oxygen atom of glycine are bridged by the water of crystallization through O--H ... O hydrogen bonds, resulting in the relatively rare trans conformation [chi = -175.7(2) degrees] for this side-chain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225786 TI - Alpha-helix structure of parathyroid hormone fragment (1-34) predicted by Monte Carlo simulated annealing. AB - Tertiary structure of parathyroid hormone fragment (1-34) is predicted by the Monte Carlo simulated annealing method. Among the 20 structures obtained after completely unbiased calculations, the lowest-energy conformation exhibits two alpha-helices around residues 2-10 and 18-22. This structure agrees with the models, especially with the location of helices, deduced from experiments. In addition, the simulation supports empirical implications in the following two points. (1) The helix near the N-terminus is more stable than the C-terminal one. (2) The rest of the peptide segments are flexible and do not tend to have any definite structure. Our calculation correctly predicts only an alpha-helix, whereas previous analyses by the Chou-Fasman method leave an ambiguity between an alpha-helix and a beta-strand. PMID- 8225787 TI - [Inequalities in mortality in the barrios of Barcelona, 1983-89]. AB - This study is aimed to describe the differences in mortality among the 38 wards of the city of Barcelona for the period 1983-89. Mortality data for years 1983 to 1989 came from the death certificates. The indicators used for the 38 city wards and the 10 city districts were: Comparative Mortality Figure, Ratio, Ratio of Potential Years of Life Lost, and Life Expectancy at birth. A descriptive analysis of these indicators by wards and district is offered; a cluster analysis based on these indicators was also performed. Wards from the same district are considered homogeneous for a given indicator, when all ward's values are higher, equal or lower to district average. The ward with the most unfavourable indicators and worst situation was Montjuic, an exponent of shanty town problems until few years ago and located near the centre. Next in ranking, from worst to best situation, did appear the four wards of the city centre district, the old historical quarter. Other wards with high mortality rates were some peripheral areas, nearly all built recently such as Ciutat Meridiana, Bon Pastor and Zona Franca, and also other old wards, such as Poble Sec. Cluster analysis classified the wards with higher mortality in several different clusters, clearly split from the other more homogeneous wards. Five of 10 districts have been considered as homogeneous. This study has allowed a deeper knowledge of the geographical distribution of mortality in the city of Barcelona, until now analysed by city districts. PMID- 8225788 TI - [The tobacco habit of the population of the province of Granada]. AB - The aims of this study are to reveal the prevalence of smoking habit amongst the residents in the Granada province and to study some components of the general attitude towards the tobacco habit. The study was a cross-sectional design which is the result of a survey based on a sample of 1537 subjects (all of them being 16 years or over and residents in this province). The participants were chosen by stratified sampling which was realized in two stages: the units of the first stage were censual sections and the second ones were usual family houses. The percentage of smokers was 51.8% male and 17.5% female. The ex-smokers' prevalence was 11.1% (19.0% and men and 3.4% women). 32.5% smokers and 75.0% ex-smokers identified tobacco as a health problem. 45% of smokers had attempted to drop tobacco one or more times. The most frequent motive being the diagnosis of disease or the smoking-related symptoms. This study has revealed the high prevalence of the smoking habit in the province of Granada. PMID- 8225789 TI - [Fish consumption and coronary mortality in the general population: a meta analysis of cohort studies]. AB - In order to estimate the effect of fish intake on ischemic heart disease mortality in the general population, we performed a meta-analysis of the epidemiologic studies involving participants free of disease at baseline published on the topic. All of the 7 studies published to date were cohort studies; however, only 5 of them reported the results with enough detail to be used in a formal meta-analysis. The total number of participants in these studies was 27,656, with an average follow-up in each study between 7.5 and 25 years and a total of 1,731 coronary deaths. The combined estimate of the relative risk for an intake of 30 g/day of fish compared to no intake was 0.96 (95% CI: 0.93-1.00; P = 0.058). Due to the presence of statistically significant heterogeneity among the studies, unexplained by a priori factors, we combined the studies assuming a random effects model, obtaining a relative risk estimate of 0.92 (95% CI: 0.84 1.01; P = 0.090). These results, together with the results of the only clinical trial of fish intake performed in post-myocardial infarction patients, in which an intake of 200-400 g/week of fatty fish reduced total mortality by 29% (relative risk of intake compared to no intake 0.71; 95% CI: 0.54-0.93), suggest a moderate beneficial effect of fish intake on coronary mortality. PMID- 8225790 TI - [An evaluation of the statistical significance and calculation of the confidence interval for the standardized mortality ratio]. AB - The standardized mortality ratio (SMR) is the ratio of the number of deaths observed (D) to the number expected (E), on the basis of the mortality rates of some reference population. Several procedures have been proposed in order to test its significance and to estimate its confidence intervals. In this study, the SMR of two causes of death in 27 health areas of Castilla-La Mancha have been calculated. The significance has been evaluated by exact Poisson test and by four methods approximating the Poisson distribution by the normal: 1) a Z statistic based on the assumption that a Poisson variate with expectation E has a standard deviation equal to square root of E; 2) the Z statistic with a continuity correction; 3) a Z statistic based on the square root transformation of a Poisson variable and 4) an approximation of the exact test by Byar. Also, the confidence intervals have been estimated by exact method and by three approximate procedures: 1) by Byar; 2) by Z statistic uncorrected and 3) by the square root transformation of the Poisson distribution. With the exact methods and Byar procedure the results were very similar; therefore, using the last to testing significance and estimate the confidence intervals of SMR, is suggested. PMID- 8225791 TI - [Control over the quality of care given pregnant women in a health area]. AB - The results of a quality control of the assistance, given through a programme of pregnancy in 12 health centres in Area number 1 of Toledo are shown here. To do so, an auditory was performed at a public hospital level and in the participating health centres, unifying explicit and normative criteria, which were the base to check a sample of 484 clinic records. In the results found, the high percentage of observance of the criteria referring to the early captation and the puerperal visit (90.3% and 94.6% respectively) is outstanding and the adequate treatment of the ferropenic anemia is the most frequently unobserved criteria (53.3%). Correction measures are established and standards for the re-evaluation are fixed. We emphasize the need to involve the second assistance level as well as the respective Management in the performance of health programmes in its aims, assuming the improvement of results as a responsibility of all of them. PMID- 8225792 TI - [The control of food hygiene during the world's fair in Seville (EXPO'92)]. AB - EXPO'92 has been one of the most important mass events held in Spain during several decades. Given the size of the event and the very complex circumstances surrounding the fair, the public health services had to face a big challenge. Food-hygiene and food-safety services, in particular were organized so as to take advantage and put into practice a fully managerial approach. Relevant conclusions can be drawn from such an experience which may be valuable not only with regards to similar mass-events in the future, but also for improving food hygiene in the overall Spanish context. In this article, the positive results obtained in the field of food hygiene and food safety at EXPO'92 are directly related by the authors with the methodology which was applied, namely the hazard analysis and critical control points. It is concluded that the interinstitutional coordination achieved played a decisive role. PMID- 8225793 TI - Incidental information processing: effects of mood, sex and caffeine. AB - This experiment assessed the effects of various parameters on incidental information processing. Participants from a nonclinical population were asked to fill out mood and personality questionnaires before completing a simple explicit and implicit memory task. It was predicted that participants with more depressive symptoms would show lower implicit memory scores. A number of sex differences were revealed: data from male participants supported the prediction; however, female participants showed a negative correlation between increasing depressive mood symptoms and implicit memory; sex differences were also present along other cognitive dimensions. Socioeconomic effects and hemispheric biases were factors considered as possible explanations for these sex differences; hemispheric bias could have resulted in differential female/male strategies being used; these may have been responsible for the consistently higher female scores found throughout the experiment. It is also suggested that implicit memory might have a separate store; this may be influenced by the default/habitual preference for a particular processing strategy (unique to the individual participant and/or the presence of depressive symptoms). PMID- 8225794 TI - Lack of behavioral sensitization to repeated cocaine administration from postnatal days 1 to 10. AB - This research determined whether sensitization (or tolerance) to the behavioral effects of cocaine in rat pups would occur following repeated cocaine administration. Rats were injected daily with 20 mg/kg of cocaine HCl s.c. from postnatal day 1 to day 10, injected with saline vehicle only, or left untreated during this period. On day 11, animals from each group were challenged with either 0, .625, 1.25, or 2.50 mg/kg of cocaine and their behavioral responses were recorded. Prior cocaine treatment did not influence the acute effects of cocaine on ultrasonic vocalizations or on any observed motor responses. In contrast, the cocaine- and saline-treated pups differed in a similar manner from the untreated control group on several behavioral measures. These results indicate that the sensitizing effects of repeated cocaine administration are not manifested during the neonatal period. However, the stimulation (stress) of handling and injection may alter the subsequent responsivity of infant rats to a cocaine challenge. PMID- 8225795 TI - Sex differences in EEG coherence in normal children. AB - The idea that sex differences in the execution of different mental tasks mean a different cerebral organization met contradictory results. Sex differences in brain electrical activity were reported by various authors. The EEG coherences show functional and anatomical interrelation, so they may reveal differences in brain organization. This paper tries to demonstrate how sex differences are manifested in EEG coherences, in 42 right-handed control children, 18 girls and 24 boys, aged 7.6 to 13.3. Since the effect of age is high on EEG measurements, regression against age was calculated and eliminated. The Z-transformed coherence values were compared by Student's t-test. The results obtained showed that girls had higher right intrahemispheric coherence values than boys in all bands. These sex differences were more numerous in the theta band. The interhemispheric differences were slight and girls had higher coherences in all bands except for the alpha band, where boys had higher interhemispheric coherence than girls. These results show slight but consistent differences in the functional organization of the right hemisphere by sex and reveal the necessity to elaborate separated EEG norms for boys and girls. PMID- 8225796 TI - Enhancement of visual evoked potentials by stimulation during low prestimulus EEG stages. AB - A new algorithm for averaging of visual evoked potentials (VEPs) is introduced and compared with the conventional method. The root mean square (RMS) values of the alpha and theta band EEG activities at the vertex 1-s preceding stimulus presentation were computed: stimuli were blocked during phases of high prestimulus activity. Stimulation was applied selectively during periods of low activity. Light stimuli of 1-s duration were given to twelve healthy, voluntary subjects. The main results were: (1) The averaged VEPs at the vertex showed an increase in the N1-P2 amplitude of about 35% in comparison to experiments with conventional stimulation. This increase was in the range of 25-30% when an ISI correction was taken into account. Frontal, temporal and parietal locations showed also increases of the N1-P2 amplitude in evoked potentials. (2) VEPs were also affected qualitatively depending on the frequency content of the prestimulus activity. PMID- 8225797 TI - Reversal of electroacupuncture tolerance by CCK-8 antiserum: an electrophysiological study on pain-related neurons in nucleus parafascicularis of the rat. AB - Two varieties of neurons were found in nucleus parafascicularis (pf) of the rat: one responds to noxious stimuli with an increase in firing (pain-excited neuron, PEN), the other with a decrease in firing (pain-inhibited neuron, PIN). Electroacupuncture (EA) has been shown to suppress PEN and excite PIN, which can be taken as an electrophysiological index for EA analgesia. This effect of EA subsided after prolonged (6 h) EA stimulation, suggesting the development of tolerance to EA. Intracerebroventricular (icv) injection of CCK-8 antiserum aiming at neutralizing endogenously released CCK-8 resulted in a complete restoration of the EA effect. Normal rabbit serum was not effective. CCK-8 antiserum per se did not affect the firing pattern of the PEN or PIN in nontolerant rat. The results obtained from single neuron recording in anesthetized animals thus confirmed those obtained in intact animals using the tail flick as the end point, implying that an excess of endogenously released CCK 8 may constitute one of the mechanisms for the development of EA tolerance. PMID- 8225798 TI - Effects of memory processing on regional brain activation: cerebral blood flow in normal subjects. AB - The role of the temporal lobe in memory has been implicated in lesion studies, which have also suggested the hypothesis of greater left hemispheric involvement in verbal, and right hemispheric involvement in facial memory. We tested these hypotheses in a sample of 27 normal right-handed subjects using the 133Xenon clearance method for measuring cerebral blood flow (CBF). The CBF was measured during resting baseline, word recognition, and face recognition conditions in counterbalanced order. CBF increased during recognition compared to baseline, and for the midtemporal lobe this increase was asymmetric to the left hemisphere for words and to the right for faces. While overall CBF levels and task related increases in CBF were uncorrelated either with performance or with delta performance (excess performance relative to basal memory abilities), laterality of task-related CBF correlated with both performance indices, showing regional specificity of correlations. This neurobehavioral probe paradigm can be applied in the study of neural substrates of normal and disturbed memory. PMID- 8225799 TI - The relationship between body height extremes and the conditioned patellar tendon reflex response. AB - The purpose of this study was to determine the effects of extreme body heights on the conditioned patellar tendon reflex response in an attempt to differentiate between the contributions of supraspinal and spinal mechanisms on long-latency reflex facilitation. Unilateral and conditioned right patellar tendon reflexes were assessed in 10 extremely short males and females and 10 extremely tall males and females. The conditioning stimulus was a contralateral patellar tendon tap and the conditioning intervals were 10, 15, 25, 50, 60, 75, 100, 150, and 300 ms. There was a long-latency facilitation of quadriceps excitability beginning at the 75 ms conditioning interval regardless of the height of the subject. It is hypothesized that the similar conditioned patellar tendon recovery profiles in the extremely short subjects and the extremely tall subjects reflects the activation of a spinal polysynaptic pathway as the predominant mechanism for our long-latency reflex facilitation. PMID- 8225800 TI - Cognitive and neuropsychological response asymmetries for adults on the left right seating axis. AB - Six studies of the relationship between classroom seating and information processing were conducted with adult subjects. Findings converged to support a hemisphericity-based explanation of effects with right-side-sitters more inclined to a response style associated with the right hemisphere--perhaps, less analytical or less cautious in responding (Experiments 1-4), more "artistic" (Experiment 5), and less lateralized (Experiment 6), and vice versa for left side sitters. First, using incidental memory tasks, subjects (N = 164) with a right side seating preference (i.e., to the speaker's left side) were found, in their test-taking behavior, to respond more positively to inaccurate distractor words (Experiments 1 and 2), particularly those with an ambiguous semantic reference (Experiment 3). For different levels of processing, seating location did not influence auditory memory (Experiments 1 and 2) but right side sitters showed enhanced visual memory for accurate words (Experiment 3). In a sample of 366 subjects, males selecting right side seating were less successful on multiple choice test performance (Experiment 4), but there were no Side differences in test taking speed or review time. In a sample of 286 subjects (Experiment 5), personality measures on the Vocational Preference Inventory revealed right sitters scored higher on the "Artistic" scale and on femininity on the Masculine Feminine scale. The final study (N = 90) revealed reduced dichotic processing on verbal tasks (digits and voiced CVs) for right side sitters. Implications are discussed. PMID- 8225801 TI - Inverse relationship between the size of pattern reversal visual evoked potentials from the left brain and the degree of left-hand preference in left handed normal subjects: importance of the left brain. AB - The relation of the degree of left-hand preference to pattern reversal visual evoked potentials (VEPs) from right and left brain was studied in male left handers. The degree of the left-hand preference was assessed by the Waterloo Handedness Questionnaire. Visual stimuli consisted of black and white checkerboard patterns generated on a TV screen. VEPs were simultaneously recorded from occipital leads of the right and left hemispheres. The degree of left-hand preference was found to be inversely and significantly related to size of VEPs only from left brain, not from right brain. That is, the conduction time, amplitude, duration, and area of N1-P1 waves linearly decreased as the degree of left-hand preference increased. These results were in accord with the testosterone hypothesis of cerebral lateralization, but not compatible with the right shift theory of handedness. It was concluded that visuomotor control by the left brain would be the main biological correlate of left-hand preference with regard to sensorimotor and cognitive functions. PMID- 8225802 TI - Attention deficits disorder and drug abuse. AB - Recent research studies have attempted to explore various aspects of attention deficit disorder and substance abuser subtypes. The purpose of the present study is to evaluate the effect of drug abuse on MMPI scale score profiles of alcoholics with or without attention deficit disorder symptoms. For the alcoholic group with ADDRT versus the alcoholic group with ADDRT and drug abuse there were no significant differences between the groups. The hypothesis that the factor of drug abuse would account for MMPI score differences between alcoholics with or without ADDRT failed to be supported. Regardless of whether or not the subject groups contained drug abusers or not, the ADDRT-diagnosed groups demonstrated clinically significant levels of psychopathology on the MMPI profiles. These results would suggest that the variable of ADDRT diagnosis is probably an important factor to consider in substance abuser populations. PMID- 8225803 TI - Multiple sclerosis: the role of the pineal gland in its timing of onset and risk of psychiatric illness. 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. It has been suggested, therefore, that the clinical manifestation of MS is dependent upon having passed the pubertal period. Since pineal melatonin secretion declines from childhood to puberty and as melatonin is an immunomodulator, we have proposed that the dramatic decline in melatonin secretion just prior to the onset of the physical manifestations of puberty may disrupt immune responses resulting in either reactivation of the infective agent or in an increased susceptibility to post pubertal infection. The fall in melatonin secretion during pre-puberty may also increase the susceptibility of these patients to affective disorder which is associated with lower melatonin secretion and the presence of a phase-advance of their biological rhythms. We predicted, therefore, a higher incidence of affective disorder in patients with pubertal or post-pubertal onset of MS compared to those in whom the disease manifested later. To test this hypothesis, we studied the incidence of affective disorder in relation to age of onset of first neurological symptoms in 31 MS patients, 6 of whom manifested symptoms of MS prior to age 18 (mean = 16.8 years). All patients with pubertal onset MS and only 48% of the control group had an affective disorder. The pubertal onset patients also had a significantly lower nocturnal melatonin levels and a lower incidence of pineal calcification on CT scan. These findings thus support the hypothesis implicating the pineal gland in the timing of onset of MS and in the risk for the development of affective disorder. PMID- 8225804 TI - A framework for a process-driven common foundation programme for graduates. AB - This paper discusses some of the problems encountered in writing a shortened Common Foundation Programme in nursing for graduates, and outlines a course which takes as its starting point the particular educational needs and requirements of the student group. Thus, the first question to be addressed by the curriculum writers when designing the course was "How can we teach these students?", rather than "What can we teach them?". The resulting process-driven course is heavily influenced by the student-centred philosophy of Carl Rogers, and utilizes a variety of large- and small-group methods to facilitate the students in gradually taking responsibility for, and making decisions about, their learning needs. The paper continues with some strategies for ensuring a smooth transition from a tutor-led, syllabus-driven start to the course, to a student-led, process-driven finish for both the theoretical and clinical components, and for the assessment schedule. Finally, a student-centred approach to evaluation is briefly outlined, and the paper concludes by suggesting that the principles employed in designing and implementing this course could be successfully transferred to a wide variety of other educational settings. PMID- 8225805 TI - Patient-nurse collaboration: a comparison of patients' and nurses' attitudes in Finland, Japan, Norway, and the U.S.A. AB - While there has been an increasing emphasis on patients' participation in decisions concerning health care and nursing in the literature as an ideal, it is not clear to what extent patients and nurses assume the consumerist attitude regarding health-care decision making. With the view that attitudes people hold regarding their role in health care and nursing will primarily affect the way they behave in health-care situations, a multinational study was carried out to examine five sets of attitudes regarding consumerism held by patients in acute care hospitals and nurses working in them. The findings from the surveys in Finland, Japan, Norway, and the U.S.A. indicate that while the patients and the nurses in these countries tend to lean toward the consumerist perspective in their attitudes, there were significant differences in the acculturation of these attitudes among the countries and between the patients and nurses. Two different models for the explanation of attitude regarding collaborative decision making in nursing practice emerged for the patients and the nurses as groups. For both groups, however, age and the more general consumerist attitudes have a bearing on their attitudes regarding collaboration in nursing. PMID- 8225806 TI - The quantity of nursing care on wards working 8- and 12-hour shifts. AB - Interest in 12-hour nursing shifts has been renewed in response to demands for improved cost-effectiveness in the NHS, but the effects of this shift on the delivery of patient care have been unclear. This paper describes the results of a repeated-measures study of 10 wards, using activity analysis to describe patterns of care under an 8-hour compared to a 12-hour shift system. Significant reductions in the amount of direct patient care were found under the 12-hour shift, with corresponding increases in unofficial work-breaks. It is suggested that these findings, which were consistent over all study wards and throughout the whole 12-hour day, demonstrate a "pacing" effect by nurses who face 12 hours on duty. Such a detrimental effect should be a major consideration when coming to any decision to implement a 12-hour shift. PMID- 8225807 TI - Factors predicting depression among Korean-American women in New York. AB - A survey was administered to 262 women aged 35-55 years who had migrated to the U.S. within the last 10 years. The survey included measures of depression, acculturative stress, social support, self-esteem, education, income, age, and length of residence in the U.S. Hierarchical regression analyses in which demographic variables were introduced first indicated that age, education, and income were all significant predictors of depression, with age positively related to depression and education and income negatively related to depression. When acculturative stress, social support, and self-esteem were introduced on subsequent steps, only self-esteem explained significant additional variability in depression. Implications for prevention and treatment programs are discussed. PMID- 8225808 TI - A DYNAMO application of microcomputer-based simulation in health sciences teaching. AB - This paper presents a concrete application of microcomputer-based simulation in health sciences education. It shows a pharmacological system dynamics model representing graphics and numerical behaviour and relations between variables, which in addition allows interaction with students for experimentation. The work also illustrates a strategy for introducing this kind of model in the classroom, as well as the suitability of their implementation, in educational environments, using professional tools such as DYNAMO and DYNEX. Subsequent evaluation of the results and comments given by the students revealed more positive attitudes towards the discipline. PMID- 8225809 TI - Stress and coping in fathers of newborns: comparisons of planned versus unplanned pregnancy. AB - The purpose of this survey was to identify stresses and coping styles in fathers of newborns, and to explore if these vary according to pregnancy planning. Data from 69 subjects were collected using the Ireton Personal Inventory and the Jalowiec Coping Scale. The most common and least often sources of stress or worry were reported. Fathers' predominant mode of coping was problem oriented. Compared to fathers with planned pregnancies, fathers with unplanned pregnancies were found to be significantly more stressed about numerous issues as well as feeling powerless in adapting to the arrival of a new infant. Implications for practice and research are explored. PMID- 8225810 TI - Factors influencing the perception of touch by elderly nursing home residents and their health caregivers. AB - The purpose of this study was to examine which factors influence the perceptions of touch by elderly nursing home residents and their health caregivers. A random sample of residents (n = 100) and a convenience sample of health caregivers (n = 100) were selected from two nursing homes. The five variables of interest were as follows: staff status; type of touch; area of the body touched; resident's locus of control; and resident's functional level. Vignettes were developed from combinations of variables which described a touch interaction between a resident and caregiver. To determine the main effects of the five variables of interest, fractional factorial analyses of variance were performed. Resident's locus of control appeared to be the most influential factor affecting residents' and caregivers' perceptions of touch. Touch was experienced as positive when it was appropriate to the situation, did not impose greater intimacy than desired, and did not impart a condescending message. Negative touch situations often involved some procedure used in a manner considered "intimate" or when discrepant with the needs of the individual. A conceptual model of interpersonal touch is described and recommendations for further study are discussed. PMID- 8225811 TI - ICD-10. PMID- 8225812 TI - Minor psychiatric morbidity in primary health care: prevalence, nature and severity. AB - The Clinical Interview Schedule (CIS) (Goldberg et al. 1970) was applied in a study investigating the prevalence rate, nature and severity of minor psychiatric disorders among U.A.E. nationals attending a primary health care centre in Al Ain, United Arab Emirates (U.A.E.). The total prevalence rate of minor psychiatric morbidity, among the group of attenders who were interviewed, was 27.6%. The morbidity was higher among females (31.9%), than males (20.3%). The highest morbidity rate among the entire sample occurred among the 35-54 year age group. The commonest diagnoses encountered were neurotic depression (55%), anxiety-depressive states (13.3%) and anxiety states (11.7%). The distribution of diagnostic entities was similar among males and females except for anxiety depressive states which were more common in females. The prevalence of neurotic depression and anxiety-depression states were highest in the middle age group, while anxiety states were highest among the young age group. The overall severity rate (OSR) among the identified cases was found to be ranging between mild and moderate severity. PMID- 8225813 TI - Ill-health and social predictors of psychotropic drug prescription in general practice. AB - Prescriptions of psychotropic drugs among 505 consecutive attenders on a single day at 32 general practices were recorded. For 404 of them the results of a screening for psychiatric morbidity conducted by means of the General Health Questionnaire (GHQ) were available. Among attenders recognised as psychiatric cases by their general practitioners the presence of concurrent somatic problems exerted a significant effect in reducing the relative odds of prescription. A very strong and significant interaction effect between male sex and high GHQ score was detected: men scoring over the threshold on the questionnaire had a risk of receiving a psychotropic drug prescription 49 times higher than the general sample. Social problems-related factors did not show significant effects when included in multivariate models comprising ill-health risk factors. PMID- 8225814 TI - Mental health policy in practice: lessons from the All Wales Strategy Mental Illness. AB - The ultimate objective of the All Wales Strategy--Mental Illness, is to produce a community based mental health service led by multidisciplinary community mental health teams. This paper is based upon a report commissioned by the Welsh Office Social Services Inspectorate based upon a review of progress in the implementation of the strategy through scrutiny of County Joint Plans and meetings with County Joint Planning Teams. In general, we found that community multidisciplinary teamwork was not well developed throughout Wales. Insufficient attention was being paid to issues of staff development and the capacity to monitor and evaluate service developments. The need for a baseline assessment of current performance by health and social services authorities seemed inescapable, if the impact of the strategy is to be reliably and validly assessed. PMID- 8225815 TI - Schizophrenic delusions among Koreans, Korean-Chinese and Chinese: a transcultural study. AB - In this transcultural study of schizophrenic delusions among Koreans, Korean Chinese and Chinese, many delusions were shown to be different among the three groups in their frequency and content and the differences could be explained by sociocultural and political factors. Delusional themes sensitive to influence by sociocultural or political situations and changes seem to be 'family', 'love affairs', 'religious matters', 'economic matters', 'specific physical damage' and 'political themes.' Delusions about 'family', 'love affair', 'being raped', 'religious matters' and 'economic and business matters' were most frequent in Koreans. Delusions of 'blood-relatedness', 'longevity' and 'political themes' were most frequent in Korean-Chinese. Delusions of 'bloodsucking and brain or viscera extracted' and 'poison or being pricked by poisoned needle' were most prominent in Chinese. PMID- 8225816 TI - Working status and anxiety levels of urban educated women in Calcutta. AB - The primary objective of the present study was to assess the impact of out-of home employment on anxiety levels of mothers. A study group of working mothers resident in Calcutta (India) was compared with a socioeconomically similar group of non-working mothers with respect to their anxiety level, measured by the Anxiety Scale Questionnaire, in terms of the total anxiety score and its various personality components. The possible relationships between anxiety score and age of these mothers as well as their children were studied. Non-working mothers showed higher anxiety levels than their working counterparts with respect to the total anxiety score as well as its components, although the differences were statistically non-significant. The anxiety scores of non-working mothers showed increasing values with increasing age of children. This trend was absent among the working mothers. The age of these mothers was not related to their anxiety level. PMID- 8225817 TI - Asian inpatients in a district psychiatric unit: an examination of presenting features and routes into care. AB - The case notes of one hundred consecutive Asian admissions at first presentation to a distric psychiatric unit were examined. Asian admissions to Greenwich Hospital comprise a young, well educated group of predominantly Punjabi Sikhs. They present mostly through crisis services often following a domiciliary visit. General practitioners (GPs) were involved in only 34% of referrals although 54% of patients had Asian GPs. They are less likely to be compulsorily admitted than has previously been reported and very infrequently came into contact with police services. Alcoholism was found mostly in male married Sikhs. Psychotic illness accounted for 51% of presentations and neurotic depression for 18%, the latter being the largest single diagnostic category. This study raises questions regarding preferred routes into care for Asians and has service implications when considering establishment of accessible community services. PMID- 8225818 TI - Psychiatric disorders among recently-arrived Eastern Europeans seen through a US refugee counseling service. AB - OBJECTIVE: To describe the types and extent of psychiatric disorders found in a group of recently-arrived Eastern Europeans being treated by native-speaking mental health providers. PATIENTS: Forty seven Hungarians, Romanians, Czechoslovakians, Poles and Gypsies referred by voluntary or government agencies, other organizations or community members to a refugee counseling service in Seattle, Washington. Over 90% were refugees and 68% maintained this status at the intake evaluation. METHOD: Clinical, descriptive study of data collected retrospectively by chart review. RESULTS: The majority of clients had poor or fair English skills and less than half were employed. Twelve (26%) had a prior mental health problem and five (11%) a psychiatric hospitalization. The most common diagnoses were adjustment reaction in 21 (45%), marital/family problems in five (11%), and paranoid schizophrenia in five (11%). CONCLUSIONS: Psychiatric disorders were frequent in this clinical population of Eastern Europeans presenting for outpatient mental health care and may be associated with previous mental health problems. Our findings emphasize the need for sensitivity to the special risks of emigration. PMID- 8225819 TI - Mechanism of oxalocalcic renal calculi generation. AB - Based on reconsideration of the contemporary knowledge on calcium oxalate urolithiasis a feasible mechanism of calculi generation is suggested. Experimental findings and clinical aspects supporting the suggested mechanism are presented. The ways of urolithiasis treatment and areas of future research are indicated. PMID- 8225820 TI - Platinum accumulation in the kidney and liver following chemotherapy with cisplatin in humans. AB - Platinum (Pt) concentrations in the kidney and liver were determined in 12 autopsied cases following cisplatin treatment. The Pt level in the liver was generally higher than that in the kidney. Pt concentration in the liver showed a significant correlation with the total dose of cisplatin. However, no dose dependent Pt accumulation was observed in the kidney. Kidney Pt levels were correlated with the doses of cisplatin injected within a 6-month period prior to the patient's death. It is suggested that the dose of cisplatin injected during a relatively short period before the patient's death affected the Pt concentration in the kidney. PMID- 8225821 TI - Imaging studies on acute focal bacterial nephritis. AB - Two cases of acute focal bacterial nephritis (AFBN) are reported. The first patient was a 35-year-old man and the diagnosis was made by ultrasonography (US) and computed tomography (CT). The second patient was a 40-year-old man and was diagnosed by US, CT and renal biopsy. They were treated with antibiotics and showed rapid improvement in both clinical symptoms and renal masses. PMID- 8225822 TI - Renocolic fistula secondary to chronic pyelonephritis. AB - Renocolic fistula is very rare. Only slightly over 100 cases have been reported in the world literature. We present a case of renocolic fistula secondary to chronic pyelonephritis discovered intraoperatively and discuss its diagnosis and management in the light of published data. PMID- 8225823 TI - Urethral valves: unusual presentation in 14-year-old boy. AB - A 14-year-old boy had multiple prostatic calculi and marked ballooning of the prostatic urethra during micturition secondary to a type III urethral valve. There were minimal changes in the bladder and upper tract. Resection of the valve corrected the patient's symptoms and signs. PMID- 8225824 TI - "To stent" or "not to stent" less than 10 mm stones? AB - The role of JJ stent in ESWL treatment of less than 10 mm urinary calculi is still controversial. In our experience routine pretreatment ureteral stenting has a definitive prophylactic and therapeutic role. The pros definitely outweigh the cons of stent "for all". PMID- 8225825 TI - Single dose of ofloxacin treatment in women with dysuria and frequency. AB - The efficacy of single dose ofloxacin in lower urinary tract infections was investigated in this study. Fifty-one women with complaints of dysuria and frequency were treated with a single dose of 400 mg ofloxacin orally. The results of urinalysis and urine culture which were performed 4-7 days after the administration of the drug were evaluated. Thirty-eight of the 47 evaluable patients (80.85%) were asymptomatic with normal urinalysis and sterile urine, thus cured. The data of the study suggest that single dose ofloxacin may be the first treatment option in the particular group of women with uncomplicated urinary tract infection. PMID- 8225826 TI - Relationship of blood groups and transitional cell carcinoma of the bladder. AB - Blood groups of 50 patients with transitional cell carcinoma of the bladder, who were followed up for a minimum of 3 years, were analyzed. The blood group distribution of the patients reflected that of the general population. There was no relationship between blood groups and stage and grade of the tumour at initial presentation. However, patients with blood group O had worse prognosis than the others. PMID- 8225827 TI - Neoadjuvant (induction) chemotherapy in advanced bladder carcinoma. AB - Traditional approaches for the treatment of locally advanced bladder tumours may not be sufficient enough. Neoadjuvant chemotherapy, a new modality, may be beneficial by enhancing the local and systemic control of the disease. The background of this new modality and early results of various trials are discussed. PMID- 8225828 TI - Cooling effect from absorption of prewarmed irrigating fluid in transurethral prostatic resection. AB - Thirty patients were studied over 10-min periods in the course of transurethral prostatic resection with measurements of rectal temperature and fluid absorption. Glycine with or without ethanol was used as the irrigant. The changes in the body temperature as reflected in oesophageal and rectal measurements in 5 of the patients were almost identical. Seven of the 30 patients became hyponatraemic and absorbed an average of 1328 ml of glycine solution. They had a significantly greater drop in rectal temperature than patients with little or no fluid absorption. This cooling effect occurred despite prewarming of the irrigant medium to 37 degrees C, which can be explained by a decrease in fluid temperature when the bag is being emptied. It was concluded that absorption of irrigating fluid is a mechanism that promotes heat loss during transurethral operations. PMID- 8225829 TI - Bladder outflow after radical prostatectomy. AB - Postoperative bladder outflow was assessed in 150 patients subjected to radical retropubic prostatectomy for clinically localized prostate cancer. The overall mean (+/- S.E.M.) postoperative peak flow rates and residual urine volumes were 16.9 +/- 1.3 ml/s and 11.4 +/- 2.1 ml. According to flow nomograms, 22% of the patients were still obstructed postoperatively due to stricture of either the anastomosis or the urethra (12%), or to partial bladder denervation (10%). Scoring of subjective symptoms was not reliable in detecting postoperative outflow obstruction, while uroflowmetry with flow nomograms identified all obstructed cases. Radical retropubic prostatectomy can induce both mechanical and functional alterations of the distal urinary tract and a strict assessment of postoperative bladder outflow is recommended. PMID- 8225830 TI - Role of phosphate in experimental uraemia. AB - Serum, corpuscular and renal tissue phosphate (Pi) concentrations in normal and chronic renal failure rats were examined. A significant correlation was found between Pi levels and indications of renal function. In addition, there were significant relationships between phosphate levels and RBC, Hb, Hct, serum Fe, serum Al, serum Si concentrations, as well as between Pi levels, Ca levels and calcium phosphate products. These results show that Pi may be used as an indicator of renal function in uraemia, anaemia and renal calcification in uraemic state. Studies in humans to confirm these relationships would be appropriate. PMID- 8225831 TI - Effects of anti-hyperlipidaemic agent or dietary protein restriction on progressive renal deterioration in adriamycin-induced nephropathy in rats. AB - Hyperlipidaemia is strongly suggested to contribute to the pathogenesis of glomerular sclerosis. This study intends to clarify the effects of either a lipid lowering agent, lovastatin (LO) or protein restriction (PR) on adriamycin (ADR) induced progressive glomerular sclerosis in rats. During the study period both serum cholesterol and triglyceride were significantly reduced in ADR-injected rats with either lovastatin administration (ADR-LO) or protein restriction (ADR PR) compared to those without such treatment (ADR-NP). At week 22, urinary protein excretion, serum creatinine, blood urea nitrogen, glomerular sclerosis and tubulointestinal alterations, which were marked in ADR-NP, were ameliorated in both ADR-LO and ADR-PR rats. ADR-LO resulted in a body weight similar to that in ADR-NP while ADR-PR induced a marked weight loss. An antihyperlipidaemic agent, such as lovastatin, seems to be a useful tool for the prevention of renal deterioration. PMID- 8225832 TI - Propeptide levels of procollagen type IV (NC1) and III (PIIINP) in patients undergoing haemodialysis. AB - In active liver disease and different forms of renal diseases the serum levels of the propeptides of procollagen types IV (NC1) and III (PIIINP) are elevated. The propeptide levels were measured by specific radioimmunoassays in 22 patients undergoing haemodialysis. NC1 concentrations in 6 chronic haemodialysis patients were within normal range (8.2 +/- 2.0 ng/ml). As compared to normal values (8.0 +/- 3 ng/ml), PIIINP concentrations were 10-fold higher in 5 intensive care patients with multi-organ failure and acute renal failure (72.9 +/- 17.9 ng/ml) and 3-fold higher in 11 chronic dialysis patients (24.3 +/- 8.7 ng/ml). Haemodialysis itself had no effect on the elimination of PIIINP. PIIINP might be a valuable marker of acute and chronic increase of collagen turnover in patients undergoing haemodialysis. PMID- 8225833 TI - The salmon GnRH encoding gene in teleost fish. PMID- 8225834 TI - Calcium channels in the neuromuscular junction. PMID- 8225835 TI - Structure, function, and regulation of the chalcone synthase. PMID- 8225836 TI - The Sertoli-germ cell communication network in mammals. AB - As soon as scientists began to study testicular structure and function, the concept emerged that SCs and GCs communicate. We now know that the seminiferous epithelium is certainly one of the most complex tissues and that the structural and functional supports of SC-GC communication are extremely elaborate. At all stages of sexual maturation, somatic cells and GCs have developed a formidable set of communication devices that are involved in attachment, displacement, cell shaping, and cell-cell transfer of molecules and cellular materials. Some of the best morphologists since the nineteenth century have studied the anatomical basis of the SC-GC dialogue and have laid the foundations to the understanding of the spermatogenic process. Further experimental efforts are still being made. In particular, new data are emerging that have enabled scientists to go beyond the descriptional or deductive aspects and to tackle the mechanical aspects. From the functional point of view, significant progress has been made in deciphering SC-GC cell language. The unique strategic position of the SC allows this cell type to receive, integrate, and emit all the signals required for the spermatogenic process to or from the extratubular compartment (e.g., FSH, testosterone), the peritubular cells (e.g., P-Mod-S), and GCs themselves. Its location also allows it to coordinate GC activity in both the transversal and the longitudinal axes of the seminiferous tubule. The SC barrier and SC products create the physical and chemical microenvironments required for the completion of each of the different steps of spermatogenesis. In addition to the tubule fluid, the SC products directly or indirectly implicated in GC control are proteins, peptides, and steroid(s) involved in germ cell proliferation, differentiation, and metabolism; transport/binding proteins; proteases; extracellular matrix components; energy metabolites; antiproteases; and various membrane components. Sertoli cell polarization results from the existence of SC-SC occluding junctions. The products required for the mitotic phase of spermatogenesis may principally be secreted basally, whereas those required for meiotic division, spermiogenesis, and sperm cells may preferentially be secreted apically. The interaction between SC factors and GCs is mediated by GC membrane receptors and different endocytic processes. The GC secondary pathway(s) involved in SC action remains a mystery. Germ cell markers that would enable a precise assessment of SC influence are lacking. Changes in the composition of the GC complement and in GC size and shape, as well as GC divisions and migration, profoundly affect SC morphology and function.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8225837 TI - The bacterial flagellar motor. PMID- 8225838 TI - [Current pathophysiologic aspects of heart failure]. PMID- 8225839 TI - [Heart failure as genetic disease: exemplified by cardiomyopathy]. PMID- 8225840 TI - [Etiology of heart failure]. PMID- 8225841 TI - [Stepwise diagnosis of heart failure]. PMID- 8225842 TI - [Therapy of acute heart failure]. PMID- 8225843 TI - [Differential therapy of chronic heart failure]. PMID- 8225844 TI - [Heart failure and arrhythmia. Prognostic and therapeutic concepts]. PMID- 8225845 TI - [Heart failure in coronary heart disease]. PMID- 8225846 TI - [Peripheral circulation in chronic heart failure]. PMID- 8225847 TI - [37-year-old patient with septic scarlet fever, hematogenous soft tissue infection and pre-shock symptoms]. PMID- 8225848 TI - [Chronic recurrent thrombotic thrombocytopenic purpura (Moschcowitz disease). An unusual long-term course in a 43-year-old patient]. PMID- 8225849 TI - [Amiodarone]. PMID- 8225850 TI - [What is the value of the left ventricular muscle mass index (LVMI) in echocardiographic diagnosis of left heart hypertrophy?]. PMID- 8225851 TI - [Goiter]. PMID- 8225852 TI - Liver metastasis model of colon cancer in the rat: immunohistochemical characterization. AB - Inoculation of 3 x 10(4) to 3 x 10(5) CC531 colon adenocarcinoma cells into the portal vein of syngeneic WAG/Rij rats provides a reproducible animal model of colon cancer liver metastasis with macroscopically visible tumor nodules at day 25 after inoculation. In this study, the inflammatory cell response in the liver sinusoids against locally induced metastases was also investigated using immunohistochemical methods. Host immune reactive cells accumulated in the liver sinusoids of tumor-bearing livers, especially resident Kupffer cells (KC; ED1+/ED2+), and to a lesser extent pit cells (OX8+/0X19-), T lymphocytes (OX8+/OX19+) and polymorphonuclear leukocytes (PMN; HIS48+). All these cell types may be involved in the lysis of CC531 cells in the liver, although only major histocompatibility complex class II+ monocytes (3D11+/ED1+/ED2-), but not resident KC, T lymphocytes nor PMN, accumulated at the peritumoral area between liver parenchyma and tumor foci. Especially monocytes but also PMN, lymphocytes and pit cells appeared to infiltrate the stromal tumor compartment. In view of the low cell dose needed, the rat liver metastasis model of colon cancer used in this study reflects well the human, weakly immunogenic, tumor-host relationships and provides an excellent opportunity for the study of cellular reactions related to chemo- and/or immunotherapy protocols. PMID- 8225853 TI - Magnetic resonance spectroscopy detects cancer in draining lymph nodes. AB - The spread of cancer cells to draining lymph nodes is an important prognostic factor for many cancers and influences postoperative therapy in patients. Histopathology is used routinely to assess if lymph nodes contain metastases. There are, however, time and resource constraints on the volume of lymph node tissue that can be examined by the pathologist in a routine laboratory (less than 2% of each node), thus major sampling errors are possible. Conventional histopathology also relies on identifying aggregates of malignant cells for a positive diagnosis. Proton (1H) magnetic resonance (MR) spectroscopy can detect chemical changes, specifically increased levels of lactate, choline, fucose and amino acids, in lymph nodes infiltrated by cancer. Increase in lactate indicates the presence of anaerobically respiring cells, whereas choline reports that the cells are replicating. Since MR spectroscopy can identify early infiltration by malignant cells, before cell clusters are visible under the light microscope, it detects micrometastases in lymph nodes missed histopathologically. Furthermore, MR spectroscopy eliminates sampling errors since the entire lymph node is examined. PMID- 8225854 TI - Different mechanisms of extracellular matrix remodeling by fibroblasts in response to human mammary neoplastic cells. AB - Human breast tumors are often associated with a fibrotic reaction termed desmoplasia. Tumor cells may indirectly modulate the composition of the extracellular matrix by influencing fibroblast properties. They may also directly interact with collagen fibrils leading to retraction of the matrix. We have studied in vitro the influence of various human mammary tumor cells on the proliferation rate of normal human fibroblasts and on their level of collagen synthesis, as well as their release of collagenase activity. Interactions between neoplastic cells and collagen matrix were investigated by incorporation of tumor cells in collagen gels (lattices) and measurement of their retraction. All cells tested (HBL100, SW613, SA52, MDA-MB-231, MCF7, MCF7/6, MCF7 ras, BT20 and T47D) were able to modulate the composition of the extracellular matrix by one or several of the mechanisms investigated. Our results also demonstrate an opposite regulation of collagen and collagenase production. The effects on the collagen metabolism and on fibroblast proliferation are probably mediated by soluble cytokines since they are reproduced by incubating the fibroblasts in the presence of medium conditioned by tumor cells. The desmoplastic reaction may thus result from different mechanisms dependent upon tumor cell types. PMID- 8225855 TI - Comparative study on the metastatic behavior of human tumors in nude, beige/nude/xid and severe combined immunodeficient mice. AB - The growth and metastatic behavior of human tumor cell lines were studied in nude, beige/nude/xid (bg/nu/xid) and severe combined immunodeficient (SCID) mice. One melanoma, two colon carcinomas and one renal carcinoma grew subcutaneously in the three strains of mice with no significant differences in tumor take and growth rate. One ovarian carcinoma formed a subcutaneously growing tumor only in SCID mice. Spontaneous metastases were observed only in mice with advanced subcutaneous tumors or at histological examination, but more frequently in bg/nu/xid and SCID mice than in nude mice. A375M melanoma formed more lung colonies in nude and bg/nu/xid mice than in SCID mice. HT-29LM colon carcinoma injected intravenously or intrasplenically formed more lung and liver colonies and lymph node metastases in bg/nu/xid mice than in nude and SCID mice. The treatment of SCID mice with anti-asGM1 serum depleted NK activity and enhanced the number of lung colonies by A375M melanoma and HT-29LM colon carcinoma. Bg/nu/xid or SCID mice may therefore offer some advantages for studying the malignant behavior of human solid tumors and differences may depend on the experimental conditions and on the tumor type. PMID- 8225856 TI - The differential resistance of B16 wild-type and F10 cells to mechanical trauma in vitro. AB - Following intravenous injection of B16 melanoma cells into mice, more than 99.9% of the cells are killed by a combination of rapid and slow processes: however, the F10 line of B16 mouse melanoma cells produces approximately 10 times as many pulmonary colonies as wild-type cells. We have attempted to determine the role of one rapid cancer cell-killing process, namely deformation-driven, loss of surface membrane integrity of the type occurring in capillaries, by the use of an in vitro model in which cells are filtered through 8-microns pores in polycarbonate membranes. In accord with in vivo observations, more wild-type than F10 cells were killed by filtration in vitro. The hypothesis that resistance to mechanical trauma of this type is enhanced by a small cell diameter and a high degree of surface rugosity is supported by measurements of these parameters on viable cells and electron micrographs. Differential resistance in these cells is associated to a major extent with a high degree of utilizable surface membrane excess, and to a minor extent with the smaller mean diameter of the F10 cells. Calculations, which are in accord with previous in vivo observations, indicate that most of the cells delivered to the capillary beds of target organs during hematogenous metastasis can be destroyed by rapid mechanical trauma, which is therefore implicated as one of a number of major contributors to metastatic inefficiency. PMID- 8225857 TI - Spontaneous corneal neovascularization in nude mice. PMID- 8225858 TI - Nerve growth factor promotes functional recovery of retinal ganglion cells after ischemia. AB - PURPOSE: To investigate the effect of a transient complete ischemia on the function of cat retina and to determine whether nerve growth factor (NGF), which was previously shown to enhance retinal ganglion cell (RGC) survival after optic nerve section in the adult rat, can promote recovery of retinal neurons after the ischemic insult. METHODS: Function of distal and proximal retina was assessed by recording the electroretinogram in response to both homogeneous flickering light (FERG) and contrast reversing gratings (PERG), respectively, 30 days after the induction of a 60-minute episode of ischemia. Visual evoked potentials in response to contrast reversing gratings were also recorded to evaluate visual acuity and contrast thresholds. Cell survival after ischemia was assessed in retinal whole-mounts stained with cresyl violet. Cats were intraocularly treated with NGF every other day, 3 times a week, for 30 days. Controls were treated with either phosphate buffered saline or cytochrome c. RESULTS: After ischemia, the FERG was not significantly affected. On the contrary, the PERG, visual acuity, and contrast thresholds were severely impaired. After NGF treatment, PERG response amplitudes were much less reduced compared to controls, and visual acuity and contrast thresholds were virtually normal. In addition, a larger number of presumed RGCs was present in the NGF-treated retinas compared to the cyt c-treated ones. CONCLUSIONS: The more proximally located retinal neurons, in particular RGCs, are highly vulnerable to ischemia. Intraocular NGF treatment was effective in enhancing the survival and functional recovery of these neurons. This suggests that NGF may represent a novel therapeutic agent for the treatment of ischemic ocular pathologies. PMID- 8225859 TI - Resistance of diabetic rat electroretinogram to hypoxemia. AB - PURPOSE: To investigate the mechanisms of the known electroretinographic abnormalities of diabetic rats and to explore effects of hypoxemia. METHODS: Subretinal and vitreal microelectrodes were used to isolate the retinal and retinal pigment epithelial components of the electroretinogram. Normoxic and hypoxemic recordings were taken from nine normal and six streptozotocin-diabetic, anesthetized, paralyzed, and ventilated pigmented rats. RESULTS: When inspired O2 was reduced the retinal pigment epithelial c-wave component of most of the normal rats diminished, whereas those of the diabetic rats, though initially smaller, were more resistant to the episode of hypoxemia (P = 0.0061). A similar trend was seen in other components. CONCLUSION: It is proposed that the reduced sensitivity of the diabetic electroretinogram to hypoxemia results from a reduced dependency of the diabetic retina on oxygen. This reduced dependence may follow from a shift in adenosine triphosphate production whereby oxidative phosphorylation is reduced by the high level of retinal intracellular glucose (Crabtree effect). A reduced oxygen demand would cause a transient increase in retinal PO2, leading to a reduction in retinal blood flow. The resulting chronic hypoperfusion of the retinal circulation may deprive the retina of vital, non-energy-related substances. PMID- 8225860 TI - Negative electroretinograms in retinitis pigmentosa. AB - PURPOSE: Patients with typical clinical features of retinitis pigmentosa were found to have the atypical electroretinographic finding of a negative waveform to a bright flash in the dark-adapted state. The full-field electroretinogram (ERG) was studied in seven such patients to understand better the pathophysiology. METHODS: Rod ERGs were isolated using blue and red flash stimuli in the dark adapted state. The rod ERG was assumed to be the sum of two major components, P3 and P2. A family of delayed Gaussian functions fitted to the rod a-wave intensity series was used to estimate the P3 component. The P2 component was derived by subtracting the estimated P3 component from the rod-isolated ERG. Long duration stimuli were used to elicit "on" and "off" components of the light-adapted cone ERG. Oscillatory potentials were isolated by digitally filtering cone ERGs to white flash stimuli. RESULTS: The estimated rod P3 component was reduced in amplitude in all patients. The derived P2 component of the rod ERG was present but abnormally reduced relative to the P3 component. Cone waveforms had decreased a-waves, "on" and "off" components. Many of the patients had a disproportionate reduction of the "on" compared to the "off" component. Photopic oscillatory potentials were either reduced in amplitude and delayed in timing or not detectable. CONCLUSIONS: The ERG findings in this subset of RP patients indicate there is dysfunction not only at the level of the photoreceptor outer segment but also at or proximal to the photoreceptor terminal region. PMID- 8225861 TI - Perceptual ranking versus visual evoked potentials for different local features in texture segregation. AB - PURPOSE: To validate recent reports of specific visual evoked potentials associated with preattentive texture segregation (tsVEPs) and to quantitatively compare perceptual segregation strength and tsVEP amplitudes across different local features. METHODS: Four local features were selected: crossed vs noncrossed, line segments differing in orientation by 90 degrees, U-shapes differing in orientation by 90 degrees, and U-shapes differing in orientation by 180 degrees. The two variants of each local feature were spatially arranged in a checkerboard pattern; for the first three features this led to pop-out of a "preattentive checkerboard." In seven subjects, perceptual segregation strength was assessed using ranking, and tsVEPs were recorded in these and three additional subjects. RESULTS: Statistically significant tsVEPs were obtained for the features crossed vs noncrossed and 90 degrees-line segments. Ranking results and tsVEP amplitudes were highly correlated (P < 0.001); the order of perceptual ranking and the order of tsVEP amplitudes were identical; 180 degrees-U-shapes had lowest ranking, mean tsVEP amplitude was 0.1 microV, close to noise. Line segments had maximal ranking, tsVEP amplitude was 1.5 microV. CONCLUSIONS: The results suggest that tsVEPs quantitatively reflect the activity of cortical mechanisms involved in texture segregation across various features. PMID- 8225862 TI - Automated suprathreshold screening for glaucoma: the Baltimore Eye Survey. AB - PURPOSE: To evaluate automated suprathreshold perimetric screening for glaucoma in a population-based survey of ocular disorders in east Baltimore, Maryland. METHODS: A population-based sample of persons > or = 40 years of age residing in 16 clusters was selected for an ocular screening examination that included automated suprathreshold testing with the Full Field 120 program of the Humphrey Field Analyzer. Subjects who failed the test underwent manual testing to confirm the defect. Subjects were referred for definitive examination by an ophthalmologist if they had an abnormal field, visual acuity worse than 20/30, intraocular pressure > 21 mm Hg, optic disc damage, a history of glaucoma, or shallow angles. The sensitivity and specificity of the automated visual field testing for identifying glaucoma was estimated and compared with other methods to screen for glaucoma. RESULTS: Of 5,341 subjects > or = 40 years of age who underwent a screening eye examination at neighborhood centers, 4,735 (89%) completed the automated field test. The median test time was 7.25 minutes per eye. Screening test results were abnormal in one or both eyes in 1,234 (26%) of the subjects. Kinetic perimetry was performed on 95% of these subjects, and defects were confirmed for 448 (36%) of them. Hence, 9.5% of the 4,735 subjects who completed the automated test were referred for definitive examination because the defect on automated perimetry was confirmed on manual testing. For a specificity of 90%, the sensitivity of the screening visual field test to detect glaucoma was 52% for 17 or more relative or absolute defects, higher than that of intraocular pressure at 39% for a cut-off of 20.5 mm Hg, vertical cup-to-disc ratio at 45% for a cut-off of 0.53, narrowest remaining rim width at 42% for a cut-off of 0.16, and was comparable to a combination of these and other nonfield parameters. CONCLUSION: Suprathreshold testing performed better than nonperimetry based screening tests for glaucoma. However, a number of logistical weaknesses of this visual field screening method were identified. PMID- 8225863 TI - Aging of the human photoreceptor mosaic: evidence for selective vulnerability of rods in central retina. AB - PURPOSE: Because previous studies suggested degeneration and loss of photoreceptors in aged human retina, the spatial density of cones and rods subserving the central 43 degrees of vision as a function of age was determined. METHODS: Cones and rods were counted in 27 whole mounted retinas from donors aged 27 to 90 years with macroscopically normal fundi. Photoreceptor topography was analyzed with new graphic and statistical techniques. RESULTS: Changes in cone density throughout this age span showed no consistent relationship to age or retinal location, and the total number of foveal cones was remarkably stable. In contrast, rod density decreased by 30%, beginning inferior to the fovea in midlife and culminating in an annulus of deepest loss at 0.5 to 3 mm eccentricity by the ninth decade. Space vacated by dying rods was filled in by larger rod inner segments, resulting in a similar rod coverage at all ages. At the temporal equator, cone density declined by 23%, but rods were stable throughout adulthood. CONCLUSIONS: The stability of both rod coverage and rhodopsin content despite decreasing cell number suggests plasticity of the adult rod system and that age related declines in scotopic sensitivity may be due to postreceptoral factors. There is no evidence for the massive loss of foveal cones required to explain even modest decrements in acuity, consistent with evidence that visual deficits at high photopic levels may be largely due to optical factors. Why the rods of central retina, which share a common support system and light exposure with the neighboring cones, are preferentially vulnerable to aging remains to be determined. PMID- 8225864 TI - Dietary restriction slows age pigment accumulation in the retinal pigment epithelium. AB - PURPOSE: The accumulation of age pigment, or lipofuscin, in postmitotic cells appears to be a universal feature of the aging process in animals. In mammals, the lipofuscin content of the retinal pigment epithelium (RPE) increases progressively during senescence. Dietary restriction has been shown to slow the rate at which many biologic parameters change during aging. Experiments were conducted to determine if dietary restriction alters the rate of age pigment accumulation in the RPE. METHODS: Male Wistar rats were placed on one of three dietary regimens starting at weaning. One group was fed a nutritionally complete diet ad libitum. Another group was fed the same diet but was only allowed to consume 60% as much food daily as the ad libitum group ate. The final group was fed ad libitum a nutritionally complete diet that had a lower caloric density per gram than the diets fed to the other animals primarily because of the replacement of carbohydrate with oat fiber. Ultrastructural morphometric analysis was used to determine the RPE age pigment content in the first group at 6 months of age, and in all of the groups at 18 months of age. RESULTS: Dietary restriction, achieved either by reducing total food intake or by reducing the caloric content of the diet, resulted in significant decreases in RPE lipofuscin accumulation. CONCLUSIONS: Dietary restriction provides a relatively simple means by which RPE age pigment content can be modulated. This should prove useful in assessing the role of RPE lipofuscin accumulation in age-related retinal disorders. That the oat fiber diet fed ad libitum was almost as effective as restriction of total food intake in slowing RPE age pigment accumulation indicates that the effect of restricted caloric intake is not mediated by almost constant hunger. PMID- 8225865 TI - IGF-1 enhancement of FGF-induced lens fiber differentiation in rats of different ages. AB - PURPOSE: Previously, using lens epithelial explants from neonatal rats, we showed that both insulin and IGF-1 synergistically enhance the effectiveness of FGF as an inducer of fiber differentiation. The authors aimed to determine whether IGF-1 enhances FGF-induced fiber differentiation in lens epithelial cells at various ages and, in particular, whether it can counter a marked age-related decline in responsiveness to FGF noted previously. METHODS: The effects of IGF-1 and bFGF were assessed using lens epithelial explants from neonatal, weanling, and adult rats. Fiber differentiation (after 13 days' culture) was monitored by crystallin ELISAs of explant lysates and also by immunofluorescent localization of crystallins. RESULTS: IGF alone had minimal effects. For younger rats, FGF alone enhanced the accumulation of alpha-, beta- and gamma-crystallins throughout explants, the peripheral region being more responsive than the central region. For adult rats, only the peripheral region responded; small amounts of alpha- and beta-crystallins were detected, but gamma-crystallin was not. Combining IGF with FGF induced gamma-crystallin in explants from adult rats (peripheral region) and enhanced the accumulation of all crystallins more than additively at all ages, mainly in the central region (young rats) or only in the peripheral region (adults). Including IGF with FGF prevented an age-related decline in the beta/alpha-crystallin ratio but not in the gamma/beta-crystallin ratio. CONCLUSIONS: IGF-1 enhances the bFGF-induced fiber differentiation responses of lens epithelial cells in neonatal, weanling, and adult rats, partially restoring an age-related decline in the responsiveness of lens cells to FGF. PMID- 8225866 TI - Intraoperative 5-fluorouracil for filtration surgery in the rabbit. AB - PURPOSE: Postoperative subconjunctival injections of 5-fluorouracil (5-FU) improve the success rate of filtration surgery, but there is still a 20% or greater failure rate at 1 year in pseudophakic and repeat trabeculectomy cases. The injections are inconvenient to give and may produce corneal epithelial toxicity and other side effects. An alternative method for administration of 5-FU is investigated. METHODS: A masked, randomized, prospective study was undertaken to compare bleb survival and complications in a rabbit model, after either a single intraoperative exposure to 5-FU (50 mg/ml concentration for 5 minutes) or five postoperative injections of 5-FU (5 mg injection on postoperative days 1, 3, 5, 7, and 9), or a combination of both, with controls that received only a 5 minute exposure to distilled water, intraoperatively. RESULTS: For the control eyes, all blebs were "flat" to masked grading, and intraocular pressure returned to preoperative levels by postoperative day 11. In the group receiving postoperative injections of 5-FU only, blebs were flat and the pressure normalized by postoperative day 14. Rabbits receiving intraoperative 5-FU only, or combined intraoperative and postoperative 5-FU, had consistently higher blebs than the former two groups, and some blebs survived until postoperative day 25. The mean intraocular pressure remained depressed in these groups until postoperative day 21 (P < 0.05 for days 5 through 18). A combination of intraoperative and postoperative 5-FU produced significantly higher blebs than intraoperative 5-FU alone on days 7 to 14. CONCLUSIONS: Intraoperative application of high-dose 5-FU to the filtration site either as a single 5-minute treatment, or in conjunction with postoperative injections of 5-FU greatly prolongs bleb function after filtration surgery in the rabbit. PMID- 8225867 TI - Expression of collagen I, smooth muscle alpha-actin, and vimentin during the healing of alkali-burned and lacerated corneas. AB - PURPOSES: Alkali-burned corneas can seldom heal properly to restore corneal transparency. To provide a better understanding of this devastating corneal injury, we compared the expression of collagen I, smooth muscle alpha-actin (alpha-SMA), and vimentin in lacerated and alkali-burned rabbit corneas. METHODS: A radiolabeled cDNA probe of alpha 1(I) chain was used in slot-blot hybridization to determine the levels of alpha 1(I) mRNA in alkali-burned corneas. In situ hybridization was used to identify the cell types that express the alpha 1(I) chain. Antibodies against collagen I, alpha-SMA, and vimentin were used in immunohistochemical studies to determine the tissue distribution of collagen I and to identify cells expressing alpha-SMA and vimentin. RESULTS: The levels of alpha 1(I) mRNA in alkali-burned corneas increased steadily after the alkali burn and reached a plateau within 2 weeks. One day after alkali burn, specific in situ hybridization signals were detected in stromal cells immediately surrounding the edge of the corneal injury. As the healing proceeded, the fibroblastic cells migrated into the injured stroma, and they showed positive reactions by in situ hybridization and by immunostaining with anti-collagen I probes. In alkali-burned corneas, retrocorneal membranes were formed 1 week after injury. This fibrillar membrane was stained by anti-collagen I antibody, and the fibroblastic cells in the membrane were hybridized by the 3H-labeled alpha 1(I) cDNA probe. No retrocorneal membrane was formed in the lacerated corneas, even after the injured corneas were allowed to heal for 3 weeks. The epithelial cells in the epithelial plug of lacerated corneas were positive by in situ hybridization, whereas the epithelial cells in the regenerated epithelium of alkali-burned cornea was not. Antibodies against alpha-SMA reacted with the migrating fibroblastic cells but did not react with epithelial cells or endothelial cells in the injured corneas. Anti-vimentin antibody reacted with fibroblastic cells, endothelial cells, and keratocytes in normal and injured corneas, and with the basal epithelial cells of injured corneas. CONCLUSIONS: During wound healing, the keratocytes that migrate to injured stroma transform into myofibroblasts. These myofibroblasts express high levels of alpha 1(I) mRNA, alpha-SMA, and vimentin. The healing of alkali burned corneas differ from that of lacerated corneas in that the retrocorneal membranes are formed in the former but not in the latter. In addition, the epithelial cells of alkali-burned corneas lack alpha 1(I) mRNA, whereas it is found in the epithelium of lacerated corneas. These differences may result from the persistence of inflammatory cells in the alkali-burned corneas. PMID- 8225868 TI - Reduction of capsaicin-induced ocular pain and neurogenic inflammation by calcium antagonists. AB - PURPOSE: To examine whether blockade of chemosensitivity of corneal nociceptors by Ca2+ antagonists decreases pain and irritation induced by capsaicin. METHODS: In adult rabbits, the number of lid-squeezing movements and the degree of palpebral opening, miotic response, and conjunctival vasodilation evoked by a bilateral instillation of 30 microliters of capsaicin (33 mM) were measured at different times (up to 5 hours) after the drug. Irritative responses to capsaicin in eyes pretreated with diltiazem, verapamil, or nifedipine were compared with those that received only the vehicle. Protein content in aqueous humor was also measured at the end of the experiment. RESULTS: Diltiazem at doses of 1 to 28 mM, administered 15 minutes before the application of capsaicin, significantly decreased scratching movements, conjunctival hyperemia, closure of the eye, and elevated aqueous protein concentration induced by capsaicin; however, it did not significantly reduce miosis. Nifedipine (2.8 and 10 mM) diminished the number of scratching movements but not other inflammatory parameters, whereas verapamil (2.8 and 10 mM) was totally ineffective in attenuating ocular signs of irritation produced by capsaicin. CONCLUSIONS: These results suggest that by lowering capsaicin-induced neural activity in nociceptive terminals, diltiazem decreases pain and neurogenic inflammation and may be useful as both an analgesic and an antiinflammatory agent in the eye. PMID- 8225869 TI - Transplantation of Y79 cells into rat eyes: an in vivo model of human retinoblastomas. AB - PURPOSE: To develop an in vivo model of human retinoblastoma by returning cultured Y79 retinoblastoma cells to the retinal environment of a widely available laboratory animal. In so doing, to study the survival, integration, and invasive characteristics expressed by tumoral cells grafted into an intraocular milieu from which these progenitor cells originated more than 20 years ago. METHODS: Using the retinal grafting method of Lazar and del Cerro, Y79 cells were injected under direct visualization into the subretinal space of Fischer 344 rats. The host rats included 36 animals that received daily injections of cyclosporin A and 4 that did not. All hosts were sacrificed 30 to 60 days after transplantation. RESULTS: Clinical examination showed vitreal invasion by masses of flocculent white material or the intravitreal formation of solid tumors. Histologic examination showed these formations to be outgrowths of grafted tumoral cells into the host retina and vitreal cavity. Highly anaplastic tumoral cells were also found lodged in subretinal and intraretinal locations. There were signs of continued and intense cell division within the grafts, with no indication of cell-mediated host reaction against the grafted cells. CONCLUSIONS: After intraretinal xenografting, human Y79 retinoblastoma cells retain a highly tumoral nature despite many years of in vitro propagation. When xenografted, these cells survive, grow, and express their malignancy within the retina of the common laboratory rat protected by a moderate immunosuppressive regimen. This partial immunosuppression is a requirement for the xenografts to prosper. This model offers a valuable opportunity to study in vivo the cellular and molecular biology of this and other human retinoblastomas, and it may facilitate the evaluation of antitumoral treatments. PMID- 8225870 TI - Subretinal space and vitreous cavity as immunologically privileged sites for retinal allografts. AB - PURPOSE: Because immune rejection is likely to be a major barrier to successful retinal transplantation, it is important to determine whether immune privilege for allogeneic retinal grafts is a feature of the subretinal space and vitreous cavity. METHODS: Newborn neural retinas of C57BL/6 mice were implanted into the subretinal space, vitreous cavity, or subconjunctival space of eyes of adult BALB/c (disparate from C57BL/6 at major and minor histocompatibility loci). At postimplantation day 12, the recipients were evaluated for donor-specific delayed hypersensitivity and examined clinically and histologically for evidence of rejection. RESULTS: Newborn neural retinal allografts in the subconjunctival space were destroyed by postimplantation day 12 and these recipients displayed intense donor-specific delayed hypersensitivity. By contrast, grafts in the subretinal space and vitreous cavity at postimplantation day 12 were found to be well differentiated and with no evidence of inflammation; these recipients failed to display donor-specific delayed hypersensitivity. Moreover, their spleens contained regulatory T cells that suppressed donor-specific delayed hypersensitivity in naive syngeneic recipients. CONCLUSIONS: Allogeneic newborn neural retinal grafts implanted in the subretinal space and vitreous cavity experience immune privilege and induce deviant immune responses resembling anterior chamber associated immune deviation. PMID- 8225871 TI - Differential S100 beta expression in choroidal and skin melanomas: quantitation by the polymerase chain reaction. AB - PURPOSE: S100 beta, a member of a calcium-binding protein family (S100s), is an important clinical marker for skin melanoma. In contrast, uveal melanomas appeared to express S100 beta protein less frequently and to a lesser degree. This study was performed to verify and extend this finding to the mRNA level. METHODS: A quantitative polymerase chain reaction (PCR)-based method was used. A ratio, comparing the S100 beta PCR fragment to that of beta-actin (an internal reference gene), was generated to compare S100 beta mRNA expression among samples. RESULTS: The ratios for skin melanomas (1.2 to 3.9; three tissues and two cell lines) were significantly higher than that for choroidal melanomas (0.1 to 0.63; seven of eight primary tumors and four of four cell lines). Only one choroidal melanoma biopsy had a ratio greater than 1. The PCR products from choroidal melanoma were identical in size and sequence to the S100 beta, as determined by gel electrophoresis and RNA conformational polymorphism. Because the ratios were also low in choroidal melanoma cell lines, the S100 beta phenotype appears to be genetically stable. CONCLUSION: S100 beta is differentially expressed at the RNA and protein levels by skin and choroidal melanomas, which are derived from distinct populations of melanocytes. However, choroidal melanomas expressing little or no S100 beta were significantly stained by antiserum specific for the S100 protein family. Taken together, these data suggest that choroidal melanocytes express another, perhaps even novel, S100 protein(s). PMID- 8225872 TI - Chemoattractant and neutrophil degranulation activities related to interleukin-8 in vitreous fluid in uveitis and vitreoretinal disorders. AB - PURPOSE: To investigate whether the cytokine interleukin-8 (IL-8), a strong chemoattractant and activator for neutrophils, is responsible for neutrophil infiltration and degranulation in the eye in uveitis. METHODS: IL-8 and elastase were measured with specific enzyme-linked immunoassays in vitreous fluid samples obtained from 69 patients with various uveitis entities. Vitreous fluid of nonuveitis patients and eye bank eyes served as controls. The chemotactic activity of vitreous fluid was tested with the Boyden chamber technique. RESULTS: IL-8 was detected in 45% of the vitreous fluid samples from uveitis patients and in 26% of vitreous fluid samples from nonuveitis patients. Vitreous fluid samples with IL-8 levels exceeding 100 pg/ml were chemotactic for neutrophils. This chemotactic activity could be blocked by 41% to 79% with a monoclonal anti-IL-8 antibody. Elastase levels in vitreous fluid of uveitis patients with detectable IL-8 were significantly higher than those in vitreous fluid samples with no detectable IL-8. CONCLUSION: These results indicate that IL-8 participates in the inflammatory processes in the eye by attracting and degranulating neutrophils. It is suggested that these processes contribute to the pathogenesis of tissue destruction in uveitis. PMID- 8225873 TI - Regulation of the levels of human trabecular matrix metalloproteinases and inhibitor by interleukin-1 and dexamethasone. AB - PURPOSE: The regulation of the trabecular meshwork's extracellular matrix is poorly understood and may involve a family of secreted proteinases, the matrix metalloproteinases. Because the trabecular extracellular matrix has been hypothesized to affect intraocular pressure, an evaluation was made of the ability of two cellular modulators to change the levels of matrix metalloproteinases in the medium of human trabecular meshwork organ explant cultures. METHODS: Trabecular explant cultures were exposed to recombinant human interleukin-1 alpha, dexamethasone, or combinations thereof for 72 hours and the culture medium was collected for analysis. Levels of stromelysin, the 72 kD gelatinase A and the 92 kD gelatinase B enzyme activity in this culture medium were assayed by substrate gel electrophoresis (zymography). Stromelysin and the tissue inhibitor of metalloproteinases (TIMP1) media protein levels were analyzed using immunoblots of Western transfers. RESULTS: Culture medium of unstimulated explants contains significant levels of the 72 kD gelatinase A and only low levels of the 92 kD gelatinase B, stromelysin, and TIMP1. Interleukin-1 alpha produces a dose-dependent several-fold elevation of gelatinase B, stromelysin, and TIMP1 without changing gelatinase A levels. Dexamethasone produces no significant change in gelatinase A and only small increases in stromelysin, gelatinase B, and TIMP1. When added together, dexamethasone antagonizes the interleukin-1 alpha-induced increase of stromelysin, gelatinase B, and TIMP1 in a dose-dependent manner. CONCLUSION: These modulators may be useful in analyzing the roles of this enzyme family in normal trabecular homeostasis and perhaps in the etiology of glaucoma. PMID- 8225874 TI - High glucose inhibits retinal capillary pericyte contractility in vitro. AB - PURPOSE: To study the effect of high glucose concentrations on pericyte contractility. METHODS: Bovine retinal capillary pericytes were cultured on silicone rubber sheets, which could be seen to wrinkle when a cell contracted. Cells were grown in glucose, or mannitol, in concentrations ranging from 5 to 40 mMol. Pericyte contractility was expressed as the percentage of cells wrinkling the silicone substratum. Observations were made fortnightly for 8 weeks. RESULTS: Cells grown in glucose exhibited a dose-dependent inhibition of contractility that was significantly greater than that seen with cells grown in mannitol, which were affected to a lesser extent. After returning to normoglycemic conditions for a further 4 weeks, the contractility of cells grown in lower glucose concentrations recovered partially, but cells grown in 40 mMol glucose did not recover at all. Pericyte proliferation was also impaired by the high-glucose growth medium. CONCLUSIONS: Pericyte contractility is inhibited by high glucose concentrations. This is consistent with the hypothesis that increased retinal blood flow may be a factor in the early pathogenesis of diabetic retinopathy. PMID- 8225875 TI - Insulin-induced hyperpolarization in retinal capillary pericytes. AB - PURPOSE: This study investigated the mechanism of insulin-induced membrane voltage hyperpolarization in retinal capillary pericytes, which possess electrical membrane properties typical for smooth muscle cells and are supposed to regulate retinal microcirculation by a contractile mechanism. METHODS: The mechanism of insulin-induced hyperpolarization was studied in cultured bovine retinal capillary pericytes using conventional microelectrodes. RESULTS: Resting voltage averaged -28 +/- 0.9 mV (mean +/- SEM, n = 45). Insulin (10(-9) to 10(-7) mol/l) induced a slow hyperpolarization in a dose-dependent fashion. Voltage change (delta V) was -3.1 +/- 0.4 mV (n = 14, P < 0.0001, = control) with an insulin concentration of 10(-8) mol/l. Blockade of potassium channels with Ba2+ (5 mmol/l) completely abolished the hyperpolarizing effect of insulin (n = 5). Apamin (10(-9) mol/l), a blocker of low-conductance Ca(2+)-activated potassium channels, also completely inhibited the insulin-induced hyperpolarization (n = 4). Blocking ATP-sensitive potassium channels with glibenclamide (10(-7) mol/l) did not reduce the hyperpolarizing action of insulin (delta V = -2.2 +/- 0.4 mV, n = 5, P = 0.29). Equivalent hyperpolarizations were recorded when insulin was added in the presence of ouabain (10(-4) mol/l) to inhibit the electrogenic Na+/ /K+/-ATPase (delta V = -3.5 +/- 1.0 mV, n = 4, P = 0.68). When pericytes were grown for 3 days in culture medium with elevated glucose concentrations (22.5 mmol/l), the resting membrane voltage and the insulin-induced hyperpolarization were not significantly altered. CONCLUSION: Insulin hyperpolarizes the membrane voltage of retinal pericytes probably mediated by activation of apamin-sensitive Ca(2+)-activated potassium channels. Therefore, hormonal modulation of membrane voltage by insulin might be an important factor in the regulation of pericyte contractility and retinal microcirculation under physiological conditions and in diabetes mellitus. PMID- 8225876 TI - Voltage-operated calcium channels in fresh and cultured rat retinal pigment epithelial cells. AB - PURPOSE: There is little known about the membrane properties of retinal pigment epithelial (RPE) cells with respect to calcium. The authors attempted to characterize membrane calcium channels from solitary fresh and cultured RPE cells from normal and dystrophic rat retinas. METHODS: RPE cells were enzymatically dissociated from eyes of neonatal rats of several strains, including dystrophic RCS strains. Membrane currents were recorded using the whole-cell version of the patch-clamp technique from either fresh or cultured cells. RESULTS: The authors observed sustained high-voltage-activated calcium channels that were dihydropyridine sensitive and closely resembled neuronal L-type calcium channels. The RCS-rdy+p+ strain was mainly investigated, but high-voltage-activated calcium channels were also recorded from fresh RPE cells of other rats regardless of age or strain, including RCS p+, RCS rdy+, Long Evans, Sprague Dawley, and also cultured RPE cells taken from a neonatal Long Evans strain. Low-voltage-activated calcium channels were not observed in any of these cells. CONCLUSION: Voltage operated calcium channels of the L-type are the main calcium channels present in rat RPE cells. Cultured cells retained the identical channels. The dystrophic RCS strains (studied until 17 days postnatal) also exhibited these channels. PMID- 8225877 TI - The antiproliferative effect of a transferrin-toxin on human retinal pigment epithelial cells and rabbit fibroblasts. AB - PURPOSE: To determine the effect of a rabbit transferrin conjugated to recombinant ricin A chain (Tfr-rRA) and the carboxylic ionophore monensin on proliferating and density-arrested human retinal pigment epithelial cells and rabbit dermal fibroblasts. METHODS: Cells were seeded on 24-well plates at 20,000 cells/cm2 and exposed to Tfr-rRA (0.1-10,000 ng/ml) with or without monensin (0.01 microM), and with or without human transferrin (65.7 mg/l) for 5 minutes to 7 days. Cells were studied morphologically and counted at 1, 2, 4, and 7 days. RESULTS: Tfr-rRA (10-10,000 ng/ml) killed proliferating human retinal pigment epithelial cells and rabbit dermal fibroblasts in a dose-dependent manner (p < or = 0.01) up to a maximum of 86% and 93%, respectively. In contrast, Tfr-rRA had minimal effect on density-arrested human retinal pigment epithelial cells and rabbit dermal fibroblasts. The cytotoxicity of Tfr-rRA was inhibited by the addition of human transferrin (65.7 mg/l), an effect that was partially overcome by longer treatment with Tfr-rRA. Monensin (0.01 microM) increased the cytotoxicity of Tfr-rRA by 4.8-fold over Tfr-rRA alone, shortened the onset of cell kill with Tfr-rRA from 48 to 24 hours (P = 0.04), and partially reversed the neutralizing effect of human transferrin. CONCLUSIONS: The results indicate that Tfr-rRA effectively inhibited the proliferation of human retinal pigment epithelial cells and rabbit dermal fibroblasts in vitro. The inhibitory effect could be modified by the addition of human transferrin or monensin. Thus, this ricin A chain conjugate may interrupt the proliferation of cells necessary in the pathogenesis of proliferative vitreoretinopathy. PMID- 8225878 TI - Requirement for vitamin A in long-term culture of human cornea. AB - PURPOSE: To develop an in vitro model for the study of vitamin A functions in the cornea by determining the morphologic responses to its absence and to the addition of varying concentrations in long-term (21-day) human organ culture. METHODS: Donor corneas were cultured for 21 days in an air-liquid environment designed to maintain normal corneal morphology in long-term culture. Corneas were graded by light microscopy and were further analyzed by transmission electron microscopy. RESULTS: Epithelium of human corneas cultured in the absence of vitamin A for 21 days showed characteristics of vitamin A deficiency: multilayering of squamous cells, bundling of tonofilaments to form keratofibrils, increased desmosomes, and loss of microplicae on the surface cells. The cells responded to the addition of vitamin A in concentrations ranging from 5 x 10(-9) to 5 x 10(-6) M. At the lowest concentration, cell differentiation and morphology appeared normal. Cells developed secretory morphology at higher concentrations of vitamin A, with intracellular vesicles, basement membrane overproduction, and reduced cell-cell and cell-substrate attachments. Keratocytes and endothelial cells showed increased intracellular vacuoles in the presence of higher concentrations of vitamin A. CONCLUSIONS: Cultured adult human corneas showed primary, dose-related responses to vitamin A in the absence of inflammatory, neuronal, or humoral factors. The epithelium was most sensitive to the absence of vitamin A and showed a dose-related response to additional vitamin A. All corneal cell layers showed morphologic changes at the highest concentration of vitamin A used. PMID- 8225879 TI - Regional differences in the subacute response of rabbit orbicularis oculi to bupivacaine-induced myotoxicity as quantified with a neural cell adhesion molecule immunohistochemical marker. AB - PURPOSE: This study examined the subacute myotoxic effects of injection of the local anesthetic bupivacaine on the orbicularis oculi muscle in the rabbit eyelid. In contrast to other muscles, the orbicularis oculi was resistant to injury by the usual anesthetic doses of bupivacaine when local infiltration is used. An attempt was made to assess the sensitivity of orbicularis oculi muscle to bupivacaine using a variety of increased bupivacaine concentrations and in combination with hyaluronidase. METHODS: Bupivacaine was injected into rabbit lower eyelids at a variety of doses with and without the injection of hyaluronidase before bupivacaine treatment. Muscle injury was assessed immunohistochemically using an antibody to an isoform of neural cell adhesion molecule, anti-leu-19, a molecule shown to localize on the surface of regenerating muscle fibers. The number of neural cell adhesion molecule-positive muscle fibers was assessed 4 and 7 days after bupivacaine injection to determine the number of fibers that were injured. RESULTS: When bupivacaine was injected into the lower eyelid at a dose of 1.5 mg, only 10% of the orbicularis oculi muscle was injured. The most effective injury involved either multiple injections of 3 mg bupivacaine or of hyaluronidase 20 minutes before the injection of 3 mg bupivacaine, resulting in injury of up to 58% of the muscle fibers. In all cases, the preseptal region of the orbicularis oculi showed a greater percentage of injury than the pretarsal portions of the muscle. CONCLUSIONS: Although multiple injections of bupivacaine and bupivacaine combined with hyaluronidase significantly increased the percentage of muscle cells injured, total destruction of the orbicularis oculi muscle was never seen. Neural cell adhesion molecule was a good marker for the quantification of the regenerating muscle fibers. It is proposed that the tight fasciculation of the orbicularis oculi muscle may play a role in preventing access of the local anesthetic to the individual muscle fibers. This demonstrates the relative clinical safety of local anesthetic injection into the eyelid. PMID- 8225880 TI - Prophylactic acyclovir effectively reduces herpes simplex virus type 1 reactivation after exposure of latently infected mice to ultraviolet B. AB - PURPOSE: To determine the potential efficacy and anatomic sites of action of prophylactic oral acyclovir using a murine model of ultraviolet-B-induced reactivation of herpes simplex 1 keratitis. METHODS: Latent infection with herpes simplex 1 (McKrae) was established in 80 National Institutes of Health inbred strain of mice. Forty of the mice were given acyclovir orally and the other 40 latently infected mice served as controls. Mice were exposed to 250 mJ/cm2 of ultraviolet-B radiation and killed on days 1, 2, 3, and 4 after ultraviolet-B radiation. Trigeminal ganglia and eyes from these mice were homogenized and incubated on Vero cell monolayers for recovery of reactivated virus. RESULTS: Based on the recovery of infectious virus after ultraviolet-B in treated versus control groups, acyclovir effectively reduced detectable viral reactivation at both the ocular level (P = 0.003) and the ganglionic level (P = 0.025). The numbers of viral culture-positive eye and trigeminal ganglia homogenates in the control group were 11 and 6 out of 40, respectively, compared to 1 and 0 out of 40 culture-positive eye and trigeminal ganglia homogenates in the acyclovir treated mice. Therapeutic serum levels of acyclovir were confirmed by high performance liquid chromatography. In the acyclovir-tested group, the single case of viral break-through at the ocular surface was not an acyclovir-resistant mutant. CONCLUSION: Prophylactic acyclovir effectively reduces the incidence of herpes simplex virus-1 reactivation after ultraviolet-B-induced reactivation in National Institutes of Health inbred strain of mice. PMID- 8225881 TI - Drug-induced retinal toxicity in albino rabbits: the effects of imipenem and aztreonam. AB - PURPOSE: To test the toxic action of two antibiotics, imipenem and aztreonam, on the functional and morphologic integrity of the albino rabbit retina. METHODS: Two commercial drugs were used--Tienam, which contains imipenem, and Azactam, which contains aztreonam. Different doses of these drugs were injected intravitreally. Retinal function was assessed from the electroretinogram (ERG) and the visual evoked potential (VEP). Retinal structure was examined at the light microscopic level. RESULTS: Imipenem did not affect the ERG and the VEP responses or the morphology of the retina up to a total injected dose of 0.98 mg (2 mg Tienam). Aztreonam was not toxic to the albino rabbit retina up to a total injected dose of 2.8 mg (5 mg of Azactam). Severe functional and morphologic retinal damage was seen when 10 mg of Azactam was injected. A similar degree of damage was seen when a dose of 5 mg L-arginine, an ingredient of Azactam, was injected into the vitreous. CONCLUSIONS: Imipenem and aztreonam are nontoxic to the albino rabbit retina at concentrations that are 500-fold higher than their effective dose against bacterial infection. Azactam is highly toxic at high levels (more than 10 mg injected into the vitreous). Most of the toxicity could be explained by the L-arginine content of the drug. PMID- 8225882 TI - The demonstration of human tumors on nude mice using gadolinium-labelled monoclonal antibodies for magnetic resonance imaging. AB - RATIONAL AND OBJECTIVES: The authors describe the magnetic resonance imaging (MRI) of a tumor in a nude mouse tumor model by means of an antibody-polylysine (Gd-DTPA) conjugate. METHODS: The monoclonal antibody, (MAB) RA96, was coupled to a polymeric gadolinium complex after periodate oxidation. This polymer, which consists of a polylysine backbone and complexing DTPA subunits, complexes an average of 65 gadolinium ions. RESULTS: The immunoreactivity of the conjugates was reduced by an average of only 30% compared with the unbound MAB RA96. Biodistribution studies in nude mice with subcutaneously implanted human colon carcinomas (WiDr [specific] and HT29 [control]) revealed a tissue concentration in the specific tumor of 8.72% of the dose applied per gram despite the high molecular weight of the conjugates. CONCLUSIONS: This tissue concentration is sufficient to achieve tumor-specific signal enhancement in the WiDr tumor on MRI. The problem of the high doses of protein required for MRI and possible solutions are discussed. PMID- 8225883 TI - Sonographic and computed tomography characteristics of liver ablation lesions induced by high-intensity focussed ultrasound. AB - RATIONALE AND OBJECTIVES: The authors have previously demonstrated the ability of high-intensity focused ultrasound (HIFU) to extracorporeally induce selective tissue destruction in the liver without causing damage to the intervening abdominal wall. The potential usefulness of HIFU as a noninvasive therapy for liver cancer has been suggested. This study observes sonographic and computed tomography (CT) characteristics of HIFU-ablated liver tissue in an attempt to assess the possibility of using these imaging methods to monitor the therapeutic results. METHODS: A sonoablated lesion was induced in the liver in each of 20 rabbits with a HIFU therapeutic system. Sequential imaging of the hepatic sonolesions with sonography and CT was performed up to 8 days after treatment, and the imaging patterns were correlated with the histopathology. RESULTS: Hepatic sonoablated tissue could be clearly visualized by sonography as a hypoechoic lesion. On contrast-enhanced CT, the sonolesions were depicted as nonenhanced low-density regions. There was good correlation among the sizes of sonography- and CT-depicted lesions and pathologic specimens. CONCLUSION: In this model, sonography and contrast-enhanced CT were useful imaging modalities for monitoring sonolesion evolution after HIFU treatment. PMID- 8225884 TI - Size of cortical bone and relationship to bone mineral density assessed by quantitative computed tomography image segmentation. AB - RATIONALE AND OBJECTIVES: The accuracy of the measurement of the size of cortical bone on computed tomography (CT) images of human vertebrae was evaluated using an automated contour detection and segmentation procedure. METHODS: Forty human lumbar vertebrae were scanned using 8-mm slices and an automated detection for definition of trabecular and cortical region of interest. The vertebrae were embedded in a polyester resin and 8-mm-thick midvertebral specimens were excised using a diamond circular saw. Contact radiographs of these specimens were performed and, after photograph magnification, the cortical area was measured using computerized planimetry. RESULTS: Cortical area measured on CT images was highly correlated with the area measured by planimetry on the specimens (r = .91; P < .001) with, however, a systematic over-estimation. A significant relationship was found between density and width of the cortex (r = .56; P < .001). CONCLUSIONS: Computed tomography is able to assess the size of cortical bone in human vertebrae, but a threshold detection algorithm, as used in the current study, is not adequate to obtain the precise anatomic dimensions. PMID- 8225885 TI - Comparison of penile duplex ultrasonography to pudendal arteriography. Variant penile arterial anatomy affects interpretation of duplex ultrasonography. AB - RATIONALE AND OBJECTIVES: Duplex ultrasonography is currently used as a noninvasive imaging modality for the functional evaluation of penile blood flow in impotent men. However, the accuracy of this test is controversial. The objective of this study was to determine the sensitivity and specificity of duplex ultrasonography, to assess the ultrasonographic parameters most predictive of arterial disease, and to evaluate the causes of misinterpretation of duplex ultrasound studies. METHODS: Duplex ultrasonography and pudendal arteriography were performed on 42 impotent men during a 4-year prospective study. The predictive power of the following ultrasonographic parameters were analyzed: cavernosal arterial diameter, peak blood flow velocity, and pulsations on real time ultrasonography. RESULTS: The results of both studies were concordant in 81% of the patients with a 77% sensitivity and 85% specificity for ultrasonography when using arteriography as the gold standard. Arterial pulsations and peak blood flow velocity were most predictive, whereas dilation was not helpful. Variant penile arterial anatomy was present in 83% of the patients and was the most frequent cause of misinterpretation of duplex ultrasonography. CONCLUSION: Duplex ultrasonography is a fairly reliable screening test for arterial disease in impotent men. Visualization of arterial pulsations and measurement of peak blood flow velocity are the best predictors of arterial function. Awareness of the potential arterial anomalies may help reduce misinterpretation of ultrasonography. Arteriography is necessary in any patient being considered for penile arterial surgery because of the high frequency of anatomic variations. PMID- 8225886 TI - Aortic and renal atherosclerotic calcifications seen on computed tomography of the spine. A positive predictor of hypertension. AB - RATIONALE AND OBJECTIVES: This study was undertaken to determine if there is an association between hypertension and aortic or renal atherosclerotic calcifications seen on computed tomography (CT) of the spine. METHODS: The lumbar CTs of 93 sequential patients were evaluated without knowledge of their blood pressures. Aortic and renal atherosclerosis was graded on a scale of 0-3. The patients' charts were reviewed for the presence of hypertension and diseases that might cause hypertension. RESULTS: Sixty-five patients had atherosclerotic calcifications. Forty-four of these were hypertensive. Logistic regression analysis revealed that calcifications in the left renal artery (P = .003), the right renal artery (P = .010) and the abdominal aorta (P = .001) are statistically significant positive predictors of hypertension. Renal artery calcifications are better predictors of hypertension than aortic calcifications. CONCLUSIONS: Renal and abdominal aortic calcifications are significant radiographic findings in lumbar spine CT examinations and their presence should be routinely included in the radiographic report. PMID- 8225887 TI - Modulation of the renal effects of contrast media by endothelium-derived nitric oxide in the rat. AB - RATIONALE AND OBJECTIVES: A possible involvement of endothelium derived relaxing nitric oxide (NO) in the pathogenesis of iodinated contrast media (CM)-induced nephrotoxicity was investigated in the rat. METHODS: Male rats (6 to 12 per group) were uninephrectomized. Six days later, the aorta was clamped above the renal artery and a low-osmolar contrast medium (CM), ioxaglate, was injected (1 mL/min; 3 minutes) via an aortic puncture in the single remaining kidney. Contrast medium was injected with or without the NO-synthase inhibitor L-NAME (100 mg/kg intravenously [i.v.] 5 minutes before CM). One group received L Arginine, the physiological precursor of NO (100 mg/kg i.v.), 5 minutes before L NAME. Phenylephrine (300 micrograms/kg; 30 min) was used as a vasoconstrictive NO independent control. The effects of iohexol, another low-osmolar CM, on creatinine clearance (CrCl) were also studied with and without pretreatment with L-NAME. A control group was subjected to a 3-minute renal ischemia only. Creatinine clearance and urinary N-acetyl-beta-D-glucosaminidase (NAG) excretion were determined before, and 24 and 48 hours after CM administration. Blinded histologic analysis was carried out after completion of the study. RESULTS: When administered alone, neither L-NAME nor L-arginine modified CrCl. Ioxaglate mildly but significantly decreased CrCl at 24 hours (-26.5% of preinjection value). This was similar to the effect observed in the control group subjected to ischemia only. When associated with L-NAME, ioxaglate markedly decreased CrCl (-58 + 11% at 24 hours, P < .05 vs. ioxaglate alone). A similar interaction was noted in the case of iohexol. L-NAME also markedly increased ioxaglate-induced urinary NAG excretion. Phenylephrine had a similar impact on renal function. L-arginine pretreatment reduced the increase in serum creatinine induced by L-NAME+ioxaglate (68 + 17 mumol/L vs. 175 + 59 mumol/L for L-NAME+ioxaglate; P < .05) and urinary NAG excretion. Ioxaglate alone induced only tubular epithelial vacuolization. When associated with L-NAME, this CM induced tubular and vascular lesions, as well as necrosis in the outer medulla. Such histologic effects were clearly inhibited by L-arginine. CONCLUSION: These data indicate that L-NAME, a specific inhibitor of NO-synthase, and phenylephrine, accentuate the nephrotoxicity of CM in the rat. This is consistent with results from the literature showing that CM toxicity is enhanced by renal ischemia. PMID- 8225888 TI - Neurotoxicity of nonionic low-osmolar contrast media. A receptor binding study. AB - RATIONALE AND OBJECTIVES: The use of the newest nonionic, water-soluble, low osmolar radiographic contrast media (CM) is still associated with occasional adverse reactions affecting the neural tissues. Because these CM display lipophilic potential in their interactions with biological membranes when diffusing within the brain parenchyma, they could affect neurotransmitter binding to the receptors. Two representative nonionic CM, iopamidol and iohexol, were studied to assess whether CM-related neurotoxicity derived from their interactions with specific receptors on neural membranes. METHODS: Binding assays were carried out in vitro on crude total membrane or crude synaptic membrane preparations from selected brain areas (cortex, striatum, hippocampus, cerebellum). The concentrations of CM and reference drugs that reduce specific binding of each ligand by 50% of its maximum value (IC50) were determined using radioligands to the receptors of the most common neurotransmitters in the central nervous system, including excitatory amino acids. RESULTS: Neither iopamidol nor iohexol inhibited the (3H) ligand binding to any kind of receptor up to very high concentrations (100 microM). CONCLUSIONS: The nonionic, low-osmolar CM did not influence the normal functions of neural membranes in our model. This suggests that occasional neurotoxic effects do not occur as a consequence of specific action on brain receptors. These CM may have an indirect, postmembrane site of action. PMID- 8225889 TI - Gadolinium-DTPA transplacental transfer and distribution in fetal tissue in rabbits. AB - RATIONALE AND OBJECTIVES: The authors assessed to what extent the commonly used the magnetic resonance imaging contrast agent, gadopentetate dimeglumine, crosses the placenta. METHODS: Eight pregnant rabbits in the third trimester were injected with 0.1 mmol/kg of gadopentetate dimeglumine, and killed 5, 15, 30, or 60 minutes after injection. Placental and fetal tissues were analyzed for gadolinium content. RESULTS: Placental concentrations of gadolinium were initially high (16.6 +/- 3.4 micrograms/gram) and then declined with a biexponential pattern. Initial gadolinium levels in the fetal organs were low and remained so except for the fetal kidneys, which showed increased levels of gadolinium from 4.3 +/- 1.1 micrograms/gram at 5 minutes to 6.8 +/- 1.8 micrograms/gram at 60 minutes. CONCLUSIONS: The results indicate that gadolinium does cross the rabbit placenta, and that concentrations in the placenta and the fetal urinary tract are sufficient for imaging as well as possible fetal toxicity. PMID- 8225890 TI - Cerebral embolization in rats induced by red blood cells treated with hypertonic X-ray contrast medium. AB - RATIONALE AND OBJECTIVES: Hypertonic X-ray contrast media induce morphologic changes in red blood cells (RBCs) and reduce their deformability when measured in vitro. This study investigated whether RBCs treated with hypertonic contrast media could induce circulatory as well as metabolic disturbances in vivo. METHODS: Autologous blood was mixed with an equal volume of meglumine diatrizoate (306 mgI/mL); 50 microliters of this mixture was infused into the left internal carotid artery of rats 2 minutes after mixing. Five minutes after infusion, the cerebral blood flow was determined using iodo[14C]antipyrine. 31P-magnetic resonance spectroscopy (MRS), 1H-MRS and 1H-magnetic resonance imaging (MRI) of the brain were performed before and after infusion of the mixture to study changes in energy metabolites and the integrity of the blood-brain barrier (BBB), as well as the development of edema. RESULTS: The blood flow decreased by 70% to 80% in the left cerebral cortex and caudate-putamen. 31P-MRS and 1H-MRS demonstrated derangement of oxidative phosphorylation. 1H-MRI demonstrated instant destruction of the BBB and gradual progress of edema in the left hemisphere. CONCLUSIONS: These results indicate that intracarotid infusion of RBCs treated with hypertonic contrast medium can induce embolization and subsequent ischemic damage to the brain. PMID- 8225891 TI - Preparation and evaluation of lyophilized iopromide-carrying liposomes for liver tumor detection. AB - RATIONALE AND OBJECTIVES: The objective of the present investigation was to demonstrate a new method for the production of iopromide-carrying liposomes. We studied biodistribution and elimination behavior, as well as the computed tomography (CT) liver imaging properties. METHODS: Iopromide-containing liposomes were prepared by the ethanol evaporation method and subsequently lyophilized. The resuspended liposomes were tested in rats and rabbits. RESULTS: After resuspension, liposomes with a mean diameter of 467 +/- 66 nm and an encapsulation rate of 41.5 +/- 5.5% were obtained. In rats, a marked accumulation of liposomal iopromide was found in the liver and spleen. In rabbits, complete renal elimination of iopromide within 7 days was demonstrated. At a dose of 150 mg total iodine/kg, a small tumor (< 0.5 cm) could be detected in a VX-2-bearing rabbit. CONCLUSIONS: The ethanol evaporation method proved to be suitable for the reproducible large-scale manufacture of iopromide liposomes with high encapsulation. The resuspended liposomes displayed favorable biodistribution, elimination and imaging characteristics. PMID- 8225892 TI - A three-dimensional stereotactic device for computed tomography-guided invasive diagnostic and therapeutic procedures. AB - RATIONALE AND OBJECTIVES: A three-dimensional stereotactic technique is presented as an improvement in precision needle placement for percutaneous diagnostic and therapeutic procedures. METHODS: This method uses transaxial computed tomography (CT) imaging for the selection of the optimal target path, and it employs a three dimensional stereotactic device designed to match precisely the CT parameters in a three-dimensional space. RESULTS: In selected cases, we reached targets as small as 1 to 2 cm while avoiding vital structures. CONCLUSIONS: In our hands, in selected cases, this has been a simple, safe, and accurate technique for improvement of percutaneous diagnostic and therapeutic procedures under CT guidance. PMID- 8225893 TI - Hodgkin's disease in adults. Part II. AB - The development of a secondary cancer is an often fatal and therefore devastating complication of the successful therapy of Hodgkin's disease. The risk of developing a secondary cancer is not the same for all patients, nor is it the same for all treatments. In 1993, treatment decisions are complicated because there are often a number of management approaches that can effectively eradicate Hodgkin's disease. Certain subgroups can be treated safely with more limited therapy. Patients with peripheral IA disease experience excellent survival following involved field radiotherapy alone without staging laparotomy. Early stage patients with good prognostic factors can be treated with radiotherapy or chemotherapy alone after a discussion of the short and long-term risks of both approaches. If unfavorable prognostic factors are present or if the patient has IIIA disease, we favor chemotherapy alone while others may employ combined modality therapy, a treatment strategy we reserve for patients with massive mediastinal disease. For advanced-stage disease, full-dose combination chemotherapy with a regimen that is familiar to the oncologist should be given. The addition of radiotherapy in this setting has not been shown to be of benefit. Each treatment strategy has to consider the individual patient and their likelihood of developing one of the complications, fatal or otherwise, of treatment. Although secondary complications must be considered in the initial strategy, unfounded fears about toxicity should not detract from delivery of therapy that has the greatest chance of cure while at the same time minimizing the risk to the patient. PMID- 8225894 TI - Facsimile transmission of radiographic images. Preliminary experiments with a personal computer and a fax modem board. PMID- 8225895 TI - Retroperitoneal cyst in an adult woman. PMID- 8225896 TI - Contrast medium-associated nephropathy. Recognition and management. AB - The single most important risk factor for the development of CAN is significant renal insufficiency, which correlates with a stable sCr greater than 1.5 mg/dL. Based on the outcome data summarized, avoidance of CAN should be our goal, as it causes significant deterioration of renal function in one of every four patients afflicted. Clearly, volume depletion should be eliminated before administering radiographic CM. In addition, high-risk patients should have a hydration protocol initiated before the procedure and continued for at least 2 hours postprocedure. At least one large cooperative study has reported a significant reduction in CAN when LOICM was compared with HOICM. Limiting the total volume of radiographic CM used for an individual study also seems to reduce the incidence of CAN. Although indications for invasive studies with radiographic RCM continue to expand, especially for elderly and other high-risk groups, using these suggestions as guidelines should minimize the risk of CAN while still obtaining the critical information needed to develop a clinical management plan. PMID- 8225897 TI - Cardiac ultrasound. Progress and prospects. AB - Echocardiography is unique in its role as both a routine, mature technology and an expanding, innovative field. For the foreseeable future, ultrasound will remain one of the most important methods of diagnostic cardiac imaging. PMID- 8225898 TI - Current status of radiographic contrast media. PMID- 8225899 TI - Advances in contrast echocardiography. PMID- 8225900 TI - Developments in magnetic resonance imaging. PMID- 8225901 TI - Functional imaging of the heart. Role of magnetic resonance imaging contrast media. PMID- 8225902 TI - Magnetic resonance angiography. An overview. PMID- 8225903 TI - Consideration of measurements of myocardial blood flow with positron-emission tomography. AB - The available data suggest that the different approaches permit the noninvasive quantification of regional myocardial blood flow in humans with comparable degrees of accuracy. Much of this accuracy depends on how carefully such measurements are performed and on the attention paid to detail when quantitative information is derived from the serially acquired PET images. Although absolute values for estimated flow may differ among laboratories and among techniques, consistency in analysis of data is of utmost importance. Yet, the choice of a specific measurement approach is often determined by practical considerations specific to a given laboratory. Factors are availability of a given tracer, ease of synthesis, proximity of the cyclotron to the scanner, instrument performance, as well as the type of studies performed in the same laboratory in other organs and associated needs for a given tracer. On the other hand, it is clear that the technology for quantifying blood flow in human myocardium has reached a point at which it can be applied routinely and reliably for the study of coronary circulatory physiology and pathophysiology of the human heart. What remains less certain at present is to what extent such quantitative approaches will augment the accuracy with which coronary artery disease can be diagnosed and characterized, and the effects of therapy monitored. However, it is clear that these novel approaches are likely to offer new insights into the pathophysiology of other than apparent coronary artery disease, as exemplified by the initial observations in patients with syndrome-X or hypertrophic cardiomyopathy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225904 TI - Issues in selecting a contrast agent for the cardiac catheterization laboratory. PMID- 8225905 TI - Public policy and funding issues. The role of the clinician. PMID- 8225906 TI - Public policy and funding issues. The role of the clinician from a Canadian perspective. AB - Radiologists and cardiologists may wish to consider an evolution of their roles: Begin to delegate some tasks to others so there is time to concentrate on the areas that are important in terms of the science of diagnosis. Train and empower technicians to do more; they are skilled and are capable of lightening the workload. It is important to offer outreach services to augment the training of other physicians. Many who work in referral and tertiary centers and teaching centers have a limited concept of what it is like to be in a center with limited technology. It is vital to know what is cost-effective and to promote those services. Physicians need media skills and ways to show other clinicians and consumers the importance of their work. It is also important to develop standards for the ordering of diagnostic procedures, to help referring physicians and young physicians to order effectively so that there is less need for policing. Evaluate new technology; informed physicians become partners in the choice and not the victim of the politics. Understand that the values about health care in Canada and the United States are changing. Have a flexible outlook. The world is changing very fast, and medicine needs to change with it. Innovate; try a new approach; take a risk. Continue to do what you already do best. Try to accept a share of social responsibility. If you really think inappropriate consumption of diagnostic services is occurring, try to deal with it. Cardiologists and radiologists have done innovative, creative things, and few people know about them because not everybody reads scientific journals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8225907 TI - An enzyme-linked immunosorbent assay for the measurement of human IgA antibody responses to Epstein-Barr virus membrane antigen. AB - An enzyme-linked immunosorbent assay (ELISA) which uses a recombinant truncated form of the Epstein-Barr virus (EBV) membrane antigen gp350/250 has been developed and used to measure human IgA antibody responses to that antigen. From comparisons with conventional immunofluorescence assays (IFA) for measuring IgA antibody responses to EBV viral capsid antigens, the ELISA shows comparable specificity and is approximately 4-fold more sensitive. Since IgA antibodies to EBV indicate a high risk of developing nasopharyngeal carcinoma (NPC), the described ELISA, which is more sensitive and objective than IFA, has potential for use in the diagnosis of NPC and for large-scale screening to identify individuals at risk for the development of this disease. PMID- 8225908 TI - Development of subunit preparations of cytomegalovirus antigen by aqueous extraction for immunization against cytomegalovirus infection. AB - This study has examined different methods of preparation of a subunit vaccine from cytomegalovirus (CMV)-infected MRC cells in terms of protein and DNA content, antigenicity and immunogenicity. Two preparations have been developed where CMV proteins were obtained by extraction of infected cells with water. Virus particles were removed from the preparations by ultracentrifugation and residual virus was inactivated by formaldehyde in the WUF preparations or by chloroform in the WUCh preparations. The preparations contained CMV proteins which may play a role in protective immune responses, and the preparations were antigenic as determined by immunodiffusion, ELISA and immunoblotting. Immunogenicity as evaluated in rabbits indicated that the preparations stimulated neutralizing and immunoprecipitating antibody. These results suggest that the gentle method of water extraction of viral antigens may be a useful protocol for vaccine preparation. PMID- 8225909 TI - Characterization of the genome of molluscum contagiosum virus type 1 between the genome coordinates 0.045 and 0.075 by DNA nucleotide sequence analysis of a 5.6 kb HindIII/MluI DNA fragment. AB - The complete DNA nucleotide sequence of a HindIII/MluI genomic DNA fragment (0.045-0.075 viral map units) from molluscum contagiosum virus type 1 (MCV-1) was determined. The HindIII/MluI DNA fragment comprises 5,646 bp with a base composition of 64.4% G + C and 35.6% A + T. The DNA sequence contains many perfect direct repeats. A cluster of three repetitive DNA elements R1, R2 and R3, with a complex structural arrangement was detected between nucleotide positions 1802 and 2107. The unit length (box) of the repetitive DNA sequences was found to be 6 bp (15 boxes) and 9 bp (24 boxes) for R1 and R2, respectively. The repetitive DNA element R3 is organized in fifteen boxes (15 bp) in which a unit length of R1 is combined with a unit length of R2. The arrangement of the repetition R3 within the DNA sequences of this particular region of the MCV-1 genome was found to be (5 x R3) + (2 x R2) + (1 x R3) + (6 x R2) + (1 x R3) + (1 x R2) + (8 x R3). Twenty-three open reading frames (ORFs) of 60-1,175 amino acid (AA) residues were detected. The largest ORF (number 17) comprises 1,175 AA with a predicted molecular weight of 126 kD. This ORF harbors a promoter signal which is located 21 nucleotides upstream from the start codon and is very similar to the early promoter signals known for vaccinia virus. This putative protein contains glutamine-enriched regions between AA residues 427 and 682 which show homologies to the corresponding glutamine-enriched regions of a variety of cellular genes like human transcriptional initiation factor (TFIID: TATA box factor). PMID- 8225910 TI - Effect of gamma-interferon or cycloheximide treatment on viral and c-myc transcripts in bovine-papillomavirus-type-1-transformed primary mouse fibroblasts. AB - We have studied the effect of gamma-interferon (IFN-gamma) treatment on the transcription of viral and c-myc oncogenes in bovine papillomavirus type 1 (BPV 1)-transformed mouse fibroblast cell lines. Upon IFN-gamma treatment, viral transcripts always decreased in cell lines containing episomal BPV-1 DNA, while the effect was variable in cell lines containing integrated BPV-1 DNA. Two series of tumour cell lines established by in vivo passage of B6B71 (episomal) and B6B31 J (integrated) cells via nude mice (NuTu A) into immunocompetent C57BL/6 mice (NuTuA B6Tul) also showed a decrease and an increase, respectively, in viral transcripts upon IFN-gamma treatment. IFN-gamma also reduced c-myc transcription in all cell lines derived from tumours, but increased it in the transformed cell cultures. There was selection of c-myc oncogene transcripts in all the tumour derived cell lines as compared to their transformed cell cultures. Cycloheximide treatment increased both viral and c-myc gene transcripts 3- to 20-fold in all cell lines. These results imply that IFN-gamma produced locally by immunological mechanisms may influence the expression of Papillomavirus oncogenes, and the response to IFN-gamma treatment may change when the viral DNA is integrated into the host genome. PMID- 8225912 TI - Orthostatic hypotension. PMID- 8225911 TI - Echovirus type 30 infection associated with aseptic meningitis in Nassau County, New York, USA. AB - An outbreak of echovirus type 30, primarily affecting infant under 1 year old, was documented in Nassau County, N.Y., USA. Twenty-four of the 40 infected patients were definitively diagnosed with aseptic meningitis. A positive correlation was found between the proportion of polymorphonuclear leukocytes and the overall cerebrospinal fluid white cell count. PMID- 8225914 TI - Arthritis in the elderly. PMID- 8225913 TI - Medical management of breast cancer. PMID- 8225915 TI - The medical management of prostate carcinoma. PMID- 8225916 TI - Nationwide hospital outpatient asthma survey. Asthma Society of Ireland. PMID- 8225917 TI - Methicillin-resistant Staphylococcus aureus (MRSA) PMID- 8225918 TI - Irish general practice and the human immunodeficiency virus. AB - Over a quarter (499) of general practitioners in the Republic of Ireland were sent a questionnaire on the human immunodeficiency virus (HIV) infection. Two hundred and fifty eight (51.7%) general practitioners returned completed questionnaires. Ninety six respondents (37.2%) had seen at least one HIV positive patient in their practice. In Dublin two thirds (67%) of respondents had seen HIV positive patients. A large majority (77.6%) of HIV positive people identified by the survey were attending general practitioners in the Eastern Health Board area. Most (61.2%) respondents favour the involvement of general practitioners in the future care of patients with HIV. PMID- 8225919 TI - A survey of final year medical students at University College, Galway. AB - The results of this survey of final year medical students at a provincial Irish university showed that over three-quarters came from families in which at least one parent was in a professional or semi-professional occupation. The majority of the students had repeated their Leaving Certificate examination. Most students expressed dissatisfaction with the career guidance counselling they received during their second level education. All the students came from secondary schools. While the majority expressed satisfaction with their choice of University course, approximately one-third had considered withdrawing from the course, particularly during the pre-clinical years. The main reasons given were financial difficulties, personal problems and difficulties with the course. Despite the provision of student counsellors and academic staff members to assist such students, the majority did not consult any staff member for help. Most students expected they would have to obtain employment outside of Ireland. PMID- 8225920 TI - General practitioner visiting rates in Ireland by entitlement category. AB - General practitioner visits reported by 6334 adults in a nationally representative sample of households for 1987 are analysed. Unlike Tussing's study some measures of health status are available and the results show that their omission biases the estimated impact of membership of Entitlement Category I in a regression model upwards. When they are included in the model, though, Category I membership still has a significant positive effect on visiting rates. PMID- 8225921 TI - Suicidal behaviour in a community based child and adolescent psychiatric service. AB - Referrals due to suicidal behaviour to a community based child and adolescent psychiatric service were reviewed. The demographic characteristics of the young people and their families, and the clinical response to such referrals is presented. There were ten new referrals due to suicidal behaviour in the period July 1990-June 1991. They were a seriously disturbed group of young people with significant family dysfunction. Many had been involved with other child welfare agencies prior to referral to our service. The majority were referred from general hospitals. We recommend that all young people, and their families, who exhibit suicidal behaviour should be offered evaluation by a child and adolescent psychiatric team. PMID- 8225922 TI - Emergency pulmonary embolectomy. AB - Pulmonary embolectomy maybe a life saving procedure in the unresponsive hypotensive patient following a massive pulmonary embolectomy despite optimal medical therapy. We report a successful pulmonary embolectomy in a 47 year old patient, three days post post coronary artery bypass grafting (CABG) and a new approach to the problem of caval filters in such patients. This case demonstrates the benefit of early surgery in this life threatening situation. PMID- 8225923 TI - A clinico-pathological study of 79 bronchoscopies in HIV-positive patients. AB - We investigated lung pathology in 61 HIV-positive patients who underwent 79 bronchoscopies. There were 27 cases of pneumocystis carinii pneumonia (PCP), 22 cases of pyogenic pulmonary infection and six cases of mycobacterial infection. Eleven cases of non-specific interstitial pneumonitis were found and 10 intravenous drug abusers had birefringent pulmonary talc granulomata in biopsy specimens. Women and intravenous drug abusers had higher rates of pyogenic pulmonary infection than men and other risk groups respectively. When compared with a previous survey from the same hospital using the same methodology the present study showed a change in the risk groups with 50% more intravenous drug abusers, 42% more haemophiliacs and 45% fewer patients with sexually transmitted infection. However, apparent changes in the spectrum of diseases encountered were not significant. PMID- 8225925 TI - Reliability of blood pressure readings in infancy. PMID- 8225924 TI - Expensive triplets. PMID- 8225926 TI - Attitudes of patients and relatives to mixed sex wards. PMID- 8225927 TI - Fragile X syndrome. PMID- 8225928 TI - Suicide in Ireland from 1950 to 1985. PMID- 8225929 TI - Twenty years after the Yom Kippur War: the belated recognition of war-induced psychic trauma. PMID- 8225930 TI - A comparison of the mental health systems of New York State and Israel. AB - The article compares characteristics of the residential and non-residential mental health services in Israel and in New York State and considers their influences on extent of service utilization. The data is based on special surveys covering all patients seen in public mental health services in New York State and in Israel during one week in 1986. The results indicate similar overall residential prevalence rates in both systems (1.9 per 1,000 population). Israeli males aged 45-64 show much higher residential prevalence rates than males of this group in NYS. The Israeli non-residential rates are much lower, 3.5 per 1,000 as compared to 5.5 in NYS. These differences are seen in both sexes and across all age groups. Overall diagnostic distributions show a greater percentage of patients with major affective disorder in NYS and a somewhat greater percentage of patients with the diagnosis of schizophrenia in Israel. Three Service Dependency Levels (SDL) are defined and their distribution by loci of care are presented. A greater proportion of patients belong to the highest SDL (1) in Israel as compared to NYS. Both in New York State and in Israel, the development of public mental health services is generally oriented to a preference for the non-residential, community treatment model over the residential, institutional model. However, Israel is less advanced than New York in developing its non residential psychiatric services, as reflected by the relative low rate of psychiatric referrals to community facilities and clinics. Analysis and explanation of these survey differences are presented. PMID- 8225931 TI - Community mental health services in Israel: current status and future trends in light of the Netanyahu Commission recommendations and the National Health Insurance Law. AB - The purpose of this article is to describe the current status of community mental health services and to examine the changes and difficulties projected as a result of the adoption of the Netanyahu Commission's recommendations and the implementation of the National Health Insurance Law. Although reform has been necessary for quite some time, particularly in the area of community mental health services, that fact does not obviate the need to raise certain questions concerning the ramifications of such change. In this article, we have attempted to focus on both the present and the future, in the following areas: organizational structure; budget; underlying concepts and ideology of community mental health services; target populations and professional personnel slated to implement the recommendations. PMID- 8225932 TI - Sexual behavior of homosexual and bisexual men attending an HIV testing clinic in Jerusalem 1986/7-1990. AB - We investigated whether the sexual practices of homosexual and bisexual men attending the Jerusalem AIDS Clinic had changed between 1986/7 and 1990. The responses of a group of 79 homosexual and bisexual men to a questionnaire in 1986/7 were compared to the responses of a group of 37 homosexuals/bisexuals in 1990. Consistent yet statistically insignificant differences in sexual behavior between the 1990 and 1986/7 groups were observed: More individuals were using condoms (53% vs 36% respectively), and at an earlier age. The practice of anal sex was less common (48% vs 67% respectively) and that of oral sex more common (46% vs 27% respectively). These differences were not confounded by the level of education between the two groups. Analyzed separately for homosexuals and bisexuals, the increase in condom use was more noticeable among homosexuals compared with bisexuals, while the decrease in anal sex was greater in bisexuals compared with homosexuals. These results may indicate a trend towards safer sexual behavior in this at risk population, but further studies are needed to support or refute these observations. PMID- 8225933 TI - Inattention, hyperactivity and speech delay at 2-4 years of age as a predictor for ADD-ADHD syndrome. AB - In the Jerusalem Institute for Child Development children with various developmental disorders at ages of 0-5 years are examined. Thirty-six children aged 2-4 years were examined by us and were found to have inattention, hyperactivity and speech delay with an IQ or DQ above 70 and were reexamined at 7 14 years of age. They were compared to a group of 27 control children. All children had a complete neurodevelopmental examination using the Touwen & Prechtel examination for Minor Neurological Dysfunction. They also had a Pollack tapper test for the identification of learning disabilities and the Conners parent's and teacher's hyperactivity rating scales. Of the 36 children from the research group 20 studied in special education classes because of behavioral disorders, inattention, and severe learning disabilities. They all had ADD-ADHD. There were 16 children in regular schools, of whom 9 had ADD-ADHD. In the control group only one child had ADD-ADHD. A very high number of the research group children failed in 2 or all 3 tests used in this study in comparison to controls. It seems that "soft" neurological signs with hyperactivity, inattention and speech delay may be early clinical signs of ADD-ADHD as 80% of the children with these clinical features developed ADD-ADHD during early school age. PMID- 8225934 TI - Treatment of resistant akathisia with ECT--a case report. AB - We present a case of neuroleptic induced akathisia resistant to traditional drug therapy which improved with electroconvulsive therapy. If confirmed in further cases, these findings call for well designed studies on the efficacy of this treatment, especially in nonresponders to traditional therapy or in a subgroup of patients suffering from akathisia and depression. PMID- 8225935 TI - Premedication with non-selective and M1-selective muscarinic antagonists before ECT. AB - Atropine treatment before electroconvulsive therapy (ECT) is used for two main reasons: a) to prevent transient post-ictal bradyarrhythmias due to excessive vagal tone; b) to minimize secretions within the respiratory tract. In the present study we have compared the effects of premedication using atropine, a non selective muscarinic antagonist, with biperiden, an M1-selective muscarinic antagonist. Cardiac rate and other cardiac parameters and respiratory tract secretions after ECT were compared in order to determine whether atropine effects following ECT involve central or peripheral antagonistic effects. Our results show that in preventing excessive sialorrhea following ECT, atropine works antagonistically through peripheral glandular M2 receptors whereas biperiden antagonizing M1 receptors fails to prevent sialorrhea. Our results are not conclusive concerning cardiac protective effects of atropine following ECT. None of the patients, whether premedicated by atropine or biperiden, displayed bradyarrhythmias following ECT. No significant differences were found in any of the measured cardiac parameters following ECT between atropine and biperiden premedications. Thus, the question whether atropine exerts its protective cardiac effects following ECT through central (probably M1) or peripheral M2 receptors remains open. PMID- 8225936 TI - The shortage of organs for transplantation: exploring the alternatives. PMID- 8225937 TI - Life as an intrinsic rather than instrumental good: the "spiritual" case against euthanasia. PMID- 8225938 TI - The common good, terminal illness, and euthanasia. PMID- 8225939 TI - The Netherlands: new regulations concerning euthanasia. PMID- 8225940 TI - In the matter of Carrie A. Coons. PMID- 8225941 TI - State of Michigan in the Circuit Court for the County of Oakland. PMID- 8225942 TI - Chemical and electrophysiological characterization of new peptide neurotoxins from the venom of the molluscivorous snail Conus textile neovicarius: a review. AB - Three peptide toxins exhibiting strong paralytic activity to molluscs, but with no paralytic effects on arthropods or vertebrates, were purified from the venom of the molluscivorous snail Conus textile neovicarius from the Red Sea. The amino acid sequences of these mollusc specific toxins are: TxIA, WCKQSGEMCNLLDQNCCDGYCIVLVCT (identical to the so-called 'King Kong peptide'); TxIB, WCKQSGEMCNVLDQNCCDGYCIVFVCT; TxIIA, WGGYSTYC gamma VDS gamma CCSDNCVRSYCT (gamma = gamma-carboxyglutamate). There is a similarity of the Cys framework of these toxins to that of the omega-conotoxins; however, their net negative charges, high content of hydrophobic residues, and uneven number of Cys residues in TxIIA are highly unusual for conotoxins. When assayed on isolated cultured Aplysia neurons, all three toxins induced spontaneous repetitive firing. The TxI toxins also induced a marked prolongation of the action potential duration. Voltage clamp experiments revealed that the TxI toxins alter the kinetics of the sodium current either by slowing down the rate of sodium current inactivation, or by recruiting silent sodium channels with slower activation and inactivation kinetics. The toxins shift the voltage-dependent steady-state Na+ current inactivation curve to more positive values by 6 mV. These changes are not associated with alteration in the rate of INa+ activation, in the peak INa+, or the sodium current reversal potential. TxI represents a new class of conotoxins with an unusual phylogenic specificity and may therefore be useful as a probe for the study of voltage gated sodium channels. (This review summarizes previously published papers). PMID- 8225943 TI - Calcium and neuronal plasticity. AB - The proposed involvement of free intracellular calcium concentration ([Ca]i) in neuronal plasticity is examined. While it is generally believed that a rise of [Ca]i is necessary for the triggering of long-term modification of synaptic connections, there are many unresolved issues related to this dogma; it is not entirely clear what is the source of the elevated calcium, how much of a calcium rise is sufficient to produce the synaptic potentiation, where and for how long, and what are the relevant chemical consequences of the transient rise of [Ca]i. It is generally believed that the dendritic spine is the locus of synaptic modification, yet little evidence exists to support this view. High resolution calcium imaging studies may contribute to the clarification of some key issues in the field of neuronal plasticity. PMID- 8225944 TI - A novel approach to correlative studies of neuronal structure and function. AB - We have recently organized and directed a consortium of manufacturers to assemble a unique electron microscopic-video-computer resource that reduces the time and effort required to make 3-dimensional reconstructions of neurons, and to collect and analyze data on synaptic organization by many fold. With the introduction of this system the quantitative study of synapses has entered a new age, characterized by a markedly increased efficiency that allows previously unrealistic studies to be carried out. In this paper, we present data designed to test the hypothesis that pyramidal cell types, identified by their intrinsic firing patterns, display characteristic inhibitory responses and distinctive synaptic patterns. These studies focus on the synaptic connectivity of regular spiking (RS) and intrinsically bursting (IB) neurons. Previous studies have shown that these neurons display distinct differences in their intrinsic membrane properties and in their morphologies as assessed with the light microscope. Under various conditions the synaptic responses of RS cells display marked inhibitory postsynaptic potentials, whereas most IB cells do not. This investigation is designed to determine if differences in the inhibitory responses of RS vs. IB cells are reflected in differences in the concentration of inhibitory synapses onto their somata. RS and IB neurons in rat somatosensory cortex were identified using intracellular recording and labeling, examined with the light microscope, and then serial thin sectioned prior to examination with the electron microscope. Synapses onto their somata and proximal dendrites were identified and plotted onto computer-assisted 3-D reconstructions made from the serial thin sections. Our analysis showed no significant difference in the types and concentration of synapses made onto the cell bodies and proximal dendrites of IB vs. RS neurons. Thus the differences observed in the inhibitory responses of IB vs. RS neurons cannot be explained by differences in the concentrations of synapses onto their somata. PMID- 8225945 TI - Evidence for plasticity of intrinsic horizontal connections in area 17 of the rat. AB - The potential for peripheral modification of intrinsic horizontal connections was studied in the primary visual cortex (area 17) of adult, hooded rats that underwent monocular enucleation (ME) or binocular enucleation (BE) at the day of birth. The intrinsic connections were labelled by localized tracer injections at various sites in area 17. The pattern of interhemispheric callosal connections was revealed in the same animals by extensive injections of a distinguishable tracer in the contralateral hemisphere. The results show marked and consistent changes in area 17 intrinsic connections as a result of the neonatal enucleations. Specifically, in contrast to normally reared rats in which intrinsic connections are short ranged and are organized fairly uniformly around the injection site, in ME cases intrinsic connections extend over half the width of area 17, usually targeting a second field within area 17. In BE cases, the injection sites result in the appearance of elongated tongues and patches of label that protrude in different directions from the injection site. The observed modifications of intrinsic connections are clearly related to the abnormalities induced by enucleations in the pattern of callosal connections, such that intrinsic connections tend to form either between callosally connected sites or between a-callosal sites but avoid intermixing the two. The results reveal a high level of plasticity in the developing intrinsic connections. A Hebbian rule of connectivity is proposed as the underlying process guiding the formation of horizontal intrinsic connections in area 17. PMID- 8225946 TI - Detection of neuronal periodic oscillations in the basal ganglia of normal and parkinsonian monkeys. AB - A feature-extracting program for detecting and quantifying periodic oscillations in auto- and cross-correlograms is described. The program performs like an "expert system" for extracting and evaluating neuronal periodic oscillations. A series of parameters (oscillation frequency, number of repeated cycles, modulation depth, etc.) is calculated, and used for grading (0 to 10) the periodic activity. The program has been applied to single-unit records from the basal ganglia of normal and parkinsonian (MPTP treated) monkeys, and the results have confirmed its accuracy and advantages over other methods. PMID- 8225947 TI - The effect of peripheral inositol injection on rat motor activity models of depression. AB - Myo-inositol is an important precursor in cellular second-messenger synthesis. It has been reported to be reduced in the cerebrospinal fluid of depressed patients and to reverse a specific effect of Li on rat behavior when given intracerebroventricularly (i.c.v.). However, myo-inositol enters the brain poorly when given peripherally, and its effects on normal rat brain have not been well studied. A series of experiments examined the effect of intraperitoneal inositol in high doses on locomotor activity, on amphetamine-induced hyperactivity, on apomorphine-induced stereotyped behavior, and on pilocarpine-induced behaviors in rats. In addition, the behavioral effect of i.c.v. inositol was compared to that of a control stereoisomer in untreated and lithium-treated rats. Few effects of peripheral myo-inositol were found in these models. However, peripheral inositol in high doses given to rats exhibiting Li-pilocarpine seizures showed that i.p. inositol could prevent these behaviors, confirming that peripheral inositol may have antidepressant potential in appropriate models. PMID- 8225948 TI - Autoimmunity and schizophrenia: an epiphenomenon or an etiology? AB - During the last 20 years considerable data have accumulated indicating that schizophrenia may have an autoimmune immunology. Patients with schizophrenia may have autoantibodies; in addition, cellular changes may occur that are compatible with autoimmunity, e.g., an increased number of CD5+ cells. If, indeed, schizophrenia does have an autoimmune pathogenesis, we should consider an alternative therapy based on immune system-manipulating drugs in addition to antipsychotic compounds. PMID- 8225949 TI - Dissociation of lexical and semantic aspects in word processing. AB - We present data and review studies that show a dissociation between identity and semantic priming. This dissociation appears as a result of employing certain prime tasks, adding cognitive load or following brain damage. Such a dissociation suggests that while activation of the word's representation is obligatory, the spread of activation to related concepts is not. PMID- 8225950 TI - The medieval brain and the cell doctrine. PMID- 8225951 TI - Sigmund Freud (born 1856 in the Czech part of the Austro-Hungarian empire, died 1939 in London). PMID- 8225952 TI - The Scottish health strategy. AB - National priorities and targets for health education in Scotland were first set in 1991. In 1992 a second policy statement--'Scotland's Health--A Challenge To Us All'--built on that foundation with a range of initiatives designed to promote health and to assist towards achievement of the national targets. Much subsequent activity has taken place to implement the initiatives. This article reports on progress. PMID- 8225953 TI - Scotland's health--a more difficult challenge for some? The price and availability of healthy foods in socially contrasting localities in the west of Scotland. AB - The recent White Paper, 'Scotland's Health: A Challenge To Us All', emphasises the importance of lifestyle factors, such as smoking, diet and exercise, in Scotland's poor health record. Targets are to be set for improvements in diet, and individuals will be encouraged to improve their eating habits. In this paper we suggest that attention should be given to the price and availability of healthy foods, particularly in socio-economically deprived areas. To illustrate the possible importance of this, we present findings from a small exploratory study of two socially contrasting and non-contiguous localities in Glasgow which indicates that price disincentives to eating healthy may be greater in poorer than in more affluent areas. PMID- 8225954 TI - An evaluation of counselling for employer-referred problem drinkers. AB - Counselling projects are facing increasing pressure from funders to evaluate their work. This study outlines the methodology adopted by the Employee Counselling Service in seeking to investigate whether the project was achieving its main aim of preventing the dismissal of clients from employment because of alcohol related problems. The results show that the project was successfully meeting this aim, with only 7.7% of their referrals being dismissed because of alcohol related problems. The evaluation also highlighted the benefits of referral for counselling prior to disciplinary procedures and gave evidence of the positive perceptions of the project held by clients and referrers. PMID- 8225955 TI - Workplace alcohol policies--are Scottish health boards exemplar employers? AB - The NHS in Scotland has been set the target of reducing the proportion of people drinking beyond the safe limits of alcohol by 20% by the year 2000. Much emphasis has been placed on development of local initiatives, including workplace alcohol policies, and guidelines were issued in 1989. In 1992 all Scottish health boards were asked for a copy of their policy and 13/15 replied (87%); 12 had policies (one in draft form only), and one did not have a policy. All regarded alcohol problems as a health issue and set out the procedures that managers should follow. However, only four addressed the use of alcohol on health board premises, and only one expressly banned drinking prior to duty and during breaks. Only four policies acknowledged the need to educate the employees about alcohol, and only three stated that the policy's effectiveness would be evaluated. This review of workplace alcohol policies shows that most Scottish health boards are not exemplar employers, and raises doubts as to whether the national target on alcohol consumption will be achieved. PMID- 8225956 TI - Perioperative deaths--detection by linkage of hospital discharge and death registration sub-files. AB - In support of the extension of the National Confidential Enquiry into Perioperative Deaths to Scotland, and in view of increased interest in the use of such deaths as clinical outcome measures in the setting of service contracts, methods have been developed employing the national Hospital Discharge (SMR1) file and Registrar General's Death Registration file on a local microcomputer for the identification of patients who die within 30 days of an operation. Direct interrogation of the SMR1 file for individual records containing both an operation date and date of death yields only those patients who die prior to discharge or transfer. Since in our study these accounted for only 52% of the total perioperative deaths, this may explain previous disillusionment with the potential of computer systems for their identification. By creating each month a SMR1 sub-file of patients having operations and linking this to a second sub-file containing the records of all patients dying in hospital, it was found that 32% of deaths occurred following transfer to a different specialty or hospital, and 8% on re-admission following discharge home. On linking the SMR1 operation sub file to the death registration file, 8% of the total perioperative deaths were shown to have taken place outside hospital. PMID- 8225957 TI - Conceptions and misconceptions about transmission of HIV/AIDS among the Scottish general public, 1988-1992. AB - Knowledge levels among the Scottish general public about so called 'casual' transmission of the AIDS virus were analysed using data collected by our Unit's survey using computer-assisted telephone interviewing methods. Five cross sections of data from the period March to May of each year were analysed and compared to provide overall estimates of knowledge on four items relating to casual transmission between respondent groups and to provide an estimate of changes over time in knowledge levels from 1988 to 1992. The data show that significant differences in knowledge exist, with the lowest levels of knowledge found among the older and among the less educated respondents. There is evidence for a continuing increase in knowledge for all items studied and for most respondent groups, but little evidence that disparities in knowledge between respondent groups are lessening over time. Misconceptions about potential risks from donating blood and kissing persist at quite high levels. PMID- 8225958 TI - Institutional care for the elderly in Scotland--rapidly changing patterns of provision. PMID- 8225959 TI - King Edward's Hospital Fund for London, 1992-93. PMID- 8225960 TI - Audit of inter-hospital transfers from a district general hospital. PMID- 8225961 TI - Variations in the Scottish Cervical Screening Programme. PMID- 8225962 TI - An examination of leadership style and its relevance to shift work in an organizational setting. AB - Results form the present research suggest that leadership style may be different for day and night and shifts, the relationship between leadership and job satisfaction is different for night and day shift employees, and the relationship between leadership and performance appraisal is not different for day and night shift workers. Implications for shift workers and further research questions are discussed. PMID- 8225963 TI - The determinants of hospital CEO compensation. AB - This study finds a significant difference in the compensation levels of public and private hospital CEOs. The difference, however, is not found to be associated with ownership structure. Rather, it is attributable to hospital characteristics such as size, location, and affiliation with a multihospital chain, and human characteristics such as gender, education, and experience. PMID- 8225964 TI - A comparative analysis of recruitment and retention of health care professionals. AB - Professional staff shortages in hospitals can drastically increase operating costs and compromise the quality of care, yet many recruitment and retention programs are ineffective. Shortages in the physical therapy and nursing professions are created by similar factors; management must also be sensitive to the differences. PMID- 8225965 TI - Reverse and double-reverse marketing for health care organizations. AB - Simple buy and sell relationships are no longer enough for health care organizations. Like other industries have done, they need to rethink and redesign their interactions with suppliers and customers in order to realize the full potential of those relationships. By establishing and managing partnerships with selected suppliers and customers, we can realize a whole new set of mutual benefits. PMID- 8225966 TI - Service guarantees in outpatient clinics: a case study. AB - It is very difficult to receive specific, actionable, constructive feedback systematically in health care delivery. This case study describes a service guarantee tool that does exactly that, at very low cost. As additional benefits, thoughtful use of service guarantees can raise morale, channel complaints that might otherwise go to the "wrong" people, and provide a unique marketing edge. PMID- 8225967 TI - Recruitment and retention of blood donors: a strategic linkage approach. AB - Through the use of discriminant analysis, 521 blood donors were analyzed to develop profiles of both high and low donors. The practical value of strategically linking recruitment and retention activities aimed at those who fit a multiple-giving profile is presented. PMID- 8225968 TI - Key decisions in marketing plan formulation for geriatric services. AB - The scope of geriatric services offered by health care institutions has expanded from the traditional hospital, long-term care, and home care services to include a variety of community services in an effort to promote the institution and provide a continuum of care for the elderly. The purposes of this article are to analyze why marketing of geriatric services is such a significant issue in health care today, to identify why a comprehensive management tool such as Service Line Management may be a key component for successful development and marketing of geriatric services, and to analyze the key decisions facing management when formulating a marketing plan. PMID- 8225969 TI - Drug-testing procedures used by hospitals: an empirical inquiry. PMID- 8225970 TI - Cost-effectiveness and data envelopment analysis. AB - Data Envelopment Analysis (DEA) identifies price and technical inefficiencies among decision-making units. With controls for differences in case-mix and standardized outcomes, DEA's "best practice" frontier can be interpreted as a "cost-effectiveness" frontier. This study illustrates the key concepts, identifies the decisions required to use the technique for medical care decision making, and presents an application to a system of nine hospitals that offer obstetric services. PMID- 8225971 TI - [The skin and the environment]. AB - The skin is in such continuous and intimate interaction with environmental influences as no other organ. Environment comes in contact with the organism in the form of substances or forces (physical, chemical, biological) and via different routes: irradiation (electromagnetic irradiation, often referred to in a simplified way as "light"), air, food, direct contact with the skin, injection, or psychosocial interactions. All these influences can lead to disease under certain conditions and in certain individuals, and especially to skin disease. The critical factors are the dose, the mode and time of contact; this is evident with regard to sunlight and its acute and chronic effects. Not only for noxious chemicals are the classification and differentiation of primary toxic reactions from hypersensitivity reactions of predisposed individuals crucial. Hypersensitivity reactions can either be immunologically mediated (allergic) or non-immunological (idiosyncrasy, pseudo-allergy). The existence of a genetic disposition does not exclude the importance of environmental factors. Atopic eczema is an example of a disease that can be influenced dramatically by a variety of environmental factors. Many toxic environmental factors are known to induce skin changes long before systemic or life-threatening diseases appear. The skin can be regarded as a "signal organ" for possible noxious environmental influences. Increased research efforts in this field are mandatory. PMID- 8225972 TI - [Siliconoma. An interdisciplinary problem]. AB - In plastic-reconstructive surgery in recent decades silicone fluid has been injected for cosmetic reasons and has occasionally led to granulomas with foreign body reaction (siliconoma). A 77-year-old woman is presented who developed protuberant facial granulomas 15 years after the injection of silicone fluid to improve scars. Histopathology revealed large lipid-filled cells, surrounded by histiocytic cells and foreign body giant cells. Laser microprobe mass analysis (LAMMA) showed peaks consistent with PO2 and PO3, which are usually found in regions of foreign body reactions. Treatment with isotretinoin given orally in low doses (20 mg total dose per day) resulted in a complete resolution of the granulomas. The anti-inflammatory action of isotretinoin was the reason for selecting this treatment modality. PMID- 8225973 TI - [Human papillomaviruses. Associated diseases of the breast]. AB - In human papillomavirus-associated acanthomas of the mamilla it was possible to detect DNA of HPV types 41, 6ma and a type related to type 16. The biology of HPV infection resembles that of anogenital lesions. Some of the HPV types found are rare or are thought possibly to have oncogenic potency. Possible evidence for link between HPV and carcinomas of the breast is discussed. PMID- 8225974 TI - [Alopecia areata and diffuse hypotrichosis associated with Ullrich-Turner syndrome. Presentation of 4 patients]. AB - Gonosomal aberrations such as Turner's syndrome are frequently associated with autoimmune diseases or with serological markers for autoimmune diseases. An autoimmune origin has been suggested for alopecia areata. The simultaneous occurrence of alopecia areata and Turner's syndrome has not formerly been reported. We present here four female patients with Turner's syndrome, who also developed typical alopecia areata. In three of these cases alopecia areata first appeared during adolescence, whilst the fourth patient showed an early manifestation at the age of 10. Two of the patients had alopecia areata of the ophiasis type, while the condition was localized in one and diffuse in the fourth. In addition, two of the patients had diffuse hypotrichosis of the scalp and reduced sweat gland activity, with no other signs of ectodermal dysplasia. The third patient had not develop body hair after her childhood, except for some sparse thin hair in the axilla and in the genital area. The coincidence of alopecia areata and Turner's syndrome may indicate some genetic relationship between the two entities. PMID- 8225975 TI - [Recurrent nodular panniculitis in alpha 1-antitrypsin deficiency. Successful dapsone therapy]. AB - We present a 59-year-old woman with chronically relapsing and necrotizing nodular panniculitis and associated alpha 1-antitrypsin deficiency. All cutaneous symptoms cleared for the first time in 10 years once she started treatment with dapsone. Chronic panniculitis should lead the clinician to determine plasma alpha 1-antitrypsin levels in more cases. PMID- 8225976 TI - [Multiple filiform hyperkeratoses]. AB - We report on a 41-year-old female patient who presented with a 3-year history of multiple filiform hyperkeratoses on the trunk. The family history revealed that her uncle had similar skin lesions. Another patient presented with filiform hyperkeratoses only on the palms of the hands and soles of the feet. This condition, referred to in English as multiple minute digitate hyperkeratoses, is a rare, mostly hereditary, disorder of cornification. As yet there is no official name for this dermatosis in German; we propose "multiple filiforme Hyperkeratosen". PMID- 8225977 TI - [Chronic mucocutaneous herpes simplex infection. Occurrence within the scope of liver-induced immunodeficiency]. AB - Herpes simplex virus (HSV) infections causing severe disease are reported frequently in patients suffering from human immunodeficiency virus (HIV) infection. This disease pattern may also be seen in an immunocompromised disease state with other causes, however, as in the case presented in this paper. An 84 year-old woman had hepatic cirrhosis resulting from chronic hepatitis C virus infection. The woman developed ulcerative lesions in and around her mouth and in the genito-anal region, and these persisted for some months. Diagnosis of HSV infection was not obtained until after extensive laboratory investigations. Aciclovir infusion therapy started immediately afterwards led to dramatic improvement of the skin and mucous membrane changes. Complete clearing of lesions was not obtained, however, because the patient died as a result of the immunosuppression. PMID- 8225978 TI - [Lymphomatoid papulosis in a child]. AB - We report a case of a 3-year-old boy who developed crops of papules and ulcerating nodules on the limbs in April 1992. Periodically, new lesions continue to erupt, while others resolve spontaneously. This course is characteristic for rhythmic paradoxical eruptions. This course and the clinical picture, supported by the histopathological and immunohistochemical findings, led to the diagnosis of lymphomatoid papulosis. Lymphomatoid papulosis is extremely rare in childhood. All published cases of lymphomatoid papulosis in children under 10 years of age are reviewed. The differential diagnosis of lymphomatoid papulosis in childhood includes arthropod assaults, pityriasis lichenoides et varioliformis acuta, primary cutaneous Hodgkin's disease, Ki-1 large cell anaplastic lymphoma and other lymphomas and pseudolymphomas in children. PMID- 8225979 TI - 1993 Distinguished Scientific Achievement Award. Presented to Robert Loevinger. PMID- 8225980 TI - 1993 Founders Award. Presented to Elizabeth Katsikas. PMID- 8225981 TI - 1993 Elda E. Anderson Award. Presented to Wesley E. Bolch. PMID- 8225982 TI - Low dose radiation carcinogenesis--issues and interpretation: the 1993 G. William Morgan Lecture. PMID- 8225983 TI - Radioactive contamination of the Techa River, the Urals. AB - The Techa River in the Urals was contaminated with high-level radioactive waste from the MAJAK nuclear installation around 1950. The total discharge to the river amounted to 100 PBq with 90Sr and 137Cs contributing approximately 10 PBq each. This study has shown that the river presently contains approximately 0.3 TBq 90Sr, > 6 TBq 137Cs, and approximately 8 GBq 239,240Pu. The estimates were made for the part of the river starting 50 km from the point of discharge and ending 240 km downstream at the confluence with the Iset River. Radioactivity was measured only in the upper 0.10-m sediments layer. The external dose rates from the contamination range from 0.1-30 microGy h-1. The activity concentrations decrease exponentially or by power functions with distance. PMID- 8225984 TI - Plutonium in daily diet in Poland after the Chernobyl accident. AB - The content of 239,240Pu was determined in the daily diet collected in northeastern Poland (Bialystok) from March 1987 to May 1992. The estimated annual intake of plutonium decreased from 774 mBq y-1 in the first year after the Chernobyl accident down to approximately 90 mBq y-1 in the sixth year. Large fluctuations of daily intake in the first 2 y suggested that a large fraction of plutonium in the daily diet originated from the external contamination. Assuming the fractional absorption factor of 10(-5) for externally contaminated foodstuffs and 10(-3) for root uptake, the assimilated 239,240Pu would be in the range from approximately 0.02 mBq up to approximately 2 mBq for the period studied. PMID- 8225985 TI - Measurement of the equilibrium factor for 222Rn progeny in Bangladesh dwellings. AB - Time-integrated solid-state nuclear track detectors were employed for the measurement of radon concentration and potential alpha-energy exposure level. From these values, the equilibrium factor F between radon and its progeny was estimated for each house. An average value of 0.40 was obtained. The experimentally and theoretically determined F values were compared. PMID- 8225986 TI - Alpha-particle dose to the liver and spleen tissues of Japanese Thorotrast patients. AB - We set out to establish an appropriate and convenient method for calculating alpha-particle absorbed doses to the liver and spleen of Thorotrast patients and to estimate a representative dose rate to the liver for the whole population of surviving and deceased Thorotrast patients in Japan. First, we determined steady state activity ratios of 232Th progeny from 13 autopsy cases and found them to be identical to those reported in German subjects. Second, we estimated the alpha dose rates in 206 subjects at autopsy from radioactivity measurements and terminal weights of the organs. Combining these results with measurements of exhaled 220Rn in survivors, we considered the representative dose rates at injection to be 0.22 Gy y-1. Additionally, our data suggest that deposition in the spleen has previously been overestimated. This is an error with some bearing on the evaluation of leukemia risks from alpha-particle irradiation. PMID- 8225987 TI - Low-level radium retention by the human body: a modification of the ICRP publication 20 retention equation. AB - Recently published rates of elimination of radium (in percentage of contemporary body content per year) demonstrated that human radium cases with low body contents lost radium more rapidly than predicted by the ICRP Publication 20 retention equation. In ICRP Publication 20, the apposition and resorption rate in compact bone is defined as 2.5% y-1, but in the derivation of the retention equation for radium a value of only 1.5% y-1 was used. In the present work, the retention equation was reevaluated by using the rate of 2.5% y-1. The revised equation was a good fit to the data for measured values of the body content of radium from environmental levels of radium in food and water. The equation also described the observed rate of loss of radium at long times after deposition for radium cases with low measured body burdens. PMID- 8225988 TI - Skin-contact exposure to tritium-gas-contaminated stainless-steel surfaces. AB - One of the radiation protection problems potentially encountered in tritium handling facilities is contamination of metal surfaces. Experiments with hairless rats have demonstrated that when intact skin is brought into contact with tritium gas-contaminated stainless-steel surfaces, tritium can be fixed as organically bound tritium (OBT) and as tritiated oxide (HTO) in the skin. The radiological hazard associated with this route of tritium uptake is determined by the retention and distribution of tritium in the skin and other organs. The experimental data suggest that the OBT in the skin serves as an input source to the rest of the body. The urinary excretion of tritium shows a biphasic excretion for OBT and a single-phase clearance for HTO from the body. The results indicate that the exposure from this mode of contamination results in long retention of tritium in the skin as well as in non-uniform distribution of tritium in organs and macromolecules. This information is useful in evaluating the possible dosimetric concerns from this mode of exposure. PMID- 8225989 TI - Probability-summation model of multiple laser-exposure effects. AB - A probability-summation model is introduced to provide quantitative criteria for discriminating independent from interactive effects of multiple laser exposures on biological tissue. Data that differ statistically from predictions of the probability-summation model indicate the action of sensitizing (synergistic/positive) or desensitizing (hardening/negative) biophysical interactions. Interactions are indicated when response probabilities vary with changes in the spatial or temporal separation of exposures. In the absence of interactions, probability-summation parsimoniously accounts for "cumulative" effects. Data analyzed using the probability-summation model show instances of both sensitization and desensitization of retinal tissue by laser exposures. Other results are shown to be consistent with probability-summation. The relevance of the probability-summation model to previous laser-bioeffects studies, models, and safety standards is discussed and an appeal is made for improved empirical estimates of response probabilities for single exposures. PMID- 8225990 TI - Relationship between exposure to radon and various types of cancer. AB - Correlations are studied between average radon levels in 1600 U.S. counties and mortality rates in them from various types of cancer. By far the strongest correlation is with lung cancer, but the sign of the correlation is negative. When smoking prevalence is included in a multiple regression, the large negative correlation between radon and lung cancer is essentially unaffected. PMID- 8225991 TI - Effects of dose limits reduction on the Argentine nuclear power plants. AB - Occupational doses are evaluated in different stages of the fuel cycle and in the operation of nuclear power plants. Trends in individual dose distribution and collective doses are analyzed. The most contributive working conditions to collective dose are identified and the implications of dose limit reduction recommended by the ICRP in 1990 are assessed. It is concluded that no relevant difficulties should appear in accomplishing the new recommendations except for implementation at Atucha I, a nuclear power plant designed in the 1960s. Some options to reduce individual and collective doses in this plant are analyzed. The change of fuel channels by new ones free from cobalt is essential to get effective improvement of occupational exposures. PMID- 8225992 TI - A factorization procedure for calculations of gamma exposure from radioactive clouds. AB - A procedure is proposed to factor the integrand in the expression for dose characteristics of gamma emission of radioactive clouds. This method permits a reduction of the multiplicity of the integration for conventional models with an arbitrary collocation of a source and a receptor. This can result in more economic numerical schemes for analyzing the consequences of radioactive releases into the atmosphere. PMID- 8225993 TI - Measurement of tritium activity in soils. AB - Tritium a radionuclide can accumulate in soils as a result of releases by the nuclear power industry. Currently there are no suitable methods to measure tritium levels in soils. A relatively simple extraction method is reported. The procedure involves an extraction of soil-borne tritium with NaOH and potassium permanganate solutions followed by standard scintillation counting procedures. The method detected tritium levels in soils that averaged 0.0099 Bq g-1 soil. Soil samples spiked with tritium were very reproducible with a coefficient of variability of 5%. PMID- 8225994 TI - Evaluation of potential strontium chelators in an octanol-water system. AB - 90Sr has a physical half-life of sufficient duration to make it a potentially dangerous contaminant from nuclear accidents and radioactive wastes. In the present study, the efficacy of 16 compounds as potential chelators of strontium was tested in vitro. Strontium solubilization from strontium carbonate and its distribution in an octanol-water system (Do/w) was determined in the absence and presence of alpha-ketoglutaric acid, Kryptofix 222, ethylenglycol-bis-(beta-amino ethylether)-N,N-tetraacetic acid, diethylentriamine pentaacetic acid, Kryptofix 5, disodium chlodronate, disodium ethidronate, oxaloacetic acid, fumaric acid, D gluconic acid, succinic acid, citric acid, D,L-2,3-diaminopropionic acid, 1,1 cyclohexanediacetic acid, tartaric acid, and trans-1,2-cy-clohexanediol. Kryptofix 222 and Kryptofix 5 significantly increased solubilized strontium, suggesting strontium chelation potential. Since in previous in vivo studies both compounds were also effective in the removal of strontium following internal contamination, it is concluded that the octanol-water system may be useful screening compounds with strontium chelation potential. PMID- 8225995 TI - Potassium iodate and its comparison to potassium iodide as a blocker of 131I uptake by the thyroid in rats. AB - Potassium iodide is the preferred thyroid blocker for personnel handling radioiodine and is recommended as a prophylaxis for the population in the near field of a nuclear reactor which would be likely to be exposed to radioiodine in an accidental breach of containment. However, in hot and humid climates, this hygroscopic chemical has a poor shelf life due to hydrolytic loss of iodine vapors. On the other hand, another iodine-rich salt, potassium iodate (KIO3), is quite stable and has a much longer shelf life. The present study compares potassium iodide and KIO3 as thyroid blockers and examines the appropriate time at which they should be administered in case of radioiodine exposure. Either of the two were given in recommended dosage (100 mg stable iodine per 70 kg body weight) at -2, 0, +2, +4, +6, and +8 h after administration of tracer quantities of radioiodine (131I) to age-, weight-, and sex-matched rats. 131I uptake in thyroid was measured 24 h after its administration in the experimental animals and compared with placebo administered controls. Results suggest that KIO3 is as effective a thyroid blocking agent as potassium iodide. In comparison to controls, 24-h thyroid uptake of 131I can be substantially reduced if potassium iodide or KIO3 is given to the animals within 2-4 h after exposure to 131I. Another noteworthy observation is that KIO3 is effective even at 8 h when administered at twice the usual dosage in comparison to the single dose, which does not show appreciable thyroid blocking properties after 8 h. PMID- 8225996 TI - Experience with a routine fecal sampling program for plutonium workers. AB - A quarterly fecal sampling program was conducted at the U. S. Department of Energy's Hanford site for congruent to 100 workers at risk for an intake of plutonium oxide and other forms of plutonium. To our surprise, we discovered that essentially all of the workers were excreting detectable activities of plutonium. Further investigation showed that the source was frequent, intermittent intakes at levels below detectability by normal workplace monitoring, indicating the extraordinary sensitivity of fecal sampling. However, the experience of this study also indicated that the increased sensitivity of routine fecal sampling relative to more common bioassay methods is offset by many problems. These include poor worker cooperation; difficulty in distinguishing low-level chronic intakes from a more significant, acute intake; difficulty in eliminating interference from ingested plutonium; and difficulty in interpreting what a single void means in terms of 24-h excretion. Recommendations for a routine fecal program include providing good communication to workers and management about reasons and logistics of fecal sampling prior to starting, using annual (instead of quarterly) fecal sampling for class Y plutonium, collecting samples after workers have been away from plutonium exposure for a least 3 d, and giving serious consideration to improving urinalysis sensitivity rather than going to routine fecal sampling. PMID- 8225997 TI - Monitoring of 35S in ash of incinerated waste. AB - The National Institute of Environmental Health Sciences incinerates low-level radioactive waste containing 3H, 14C, and 35S as a means of volume reduction. The primary isotope remaining in the ash is 35S. Some effects of the chemical and physical form of the waste on sulfur retention in ash and several methods for determining the activity remaining in the ash have been explored. It was found that the sulfur in sodium sulfate has a higher retention in ash than the sulfur in methionine and that the presence of glass appears to enhance the retention. Liquid scintillation and proportional counting methods were compared. A sulfur extraction procedure using liquid scintillation counting was found to give the lowest minimum detectable activity. However, it was determined that a more practical method for routine monitoring is to suspend a small quantity of ash in a liquid scintillation gel. PMID- 8225998 TI - Comparing bone sarcoma induction in humans from 224Ra and 226+228Ra. PMID- 8225999 TI - Decay of gamma radiation in fallout following a nuclear war. PMID- 8226000 TI - The CCRN-CCMD partnership: advancing the quality of patient care. PMID- 8226001 TI - Blood pressure reactivity in the evaluation of resting blood pressure and mood responses to pindolol and propranolol in hypertensive patients. AB - OBJECTIVE: The purpose of this exploratory study was to evaluate the relationship of blood pressure reactivity during exercise to treatment responsiveness to two commonly used beta-adrenergic blocking agents, propranolol and pindolol. DESIGN: Prospective, placebo-controlled, balanced, cross-over clinical trial. SETTING: University-affiliated medical center. PATIENTS: A convenience sample of 19 white male subjects with mild to moderate essential hypertension were studied. The mean age was 63.4 years (SD = 5.2). The mean resting systolic blood pressure (SBP) was 158.6 mm Hg (SD = 12.3) and mean resting diastolic blood pressure (DBP) was 96.4 mm Hg (SD = 8.6). They had no clinical evidence of secondary hypertension, diabetes, heart, liver, pulmonary, or renal disease. OUTCOME MEASURES: Resting blood pressure; blood pressure reactivity to exercise; self-report measures of depressive symptoms, and mood disturbances. INTERVENTION: Antihypertensive medication was tapered off and subjects were free of all prescription drugs for 2 weeks. Subjects were randomly assigned to propranolol-pindolol or pindolol propranolol group. Each 4- to 6-week treatment phase was preceded by a 2-week placebo phase. At the end of the initial placebo phase and each active drug treatment phase, subjects were assessed for depression and mood disturbances by use of standardized measures and were given a graded exercise test on a cycle ergometer. Resting blood pressure was assessed weekly and before each exercise test. RESULTS: Significant relationships between DBP reactivity to exercise during the placebo phase and the degree of blood pressure and mood responsiveness to pindolol and propranolol treatment were observed. Subjects demonstrating high DBP reactivity required high doses of beta-blocker for resting DBP reduction, and these subjects showed the least change in mood at high doses. Similar patterns were found for the relationship of SBP reactivity and blood pressure and mood responsiveness to drug treatment, but these relationships were not statistically significant. CONCLUSIONS: DBP reactivity to exercise during the placebo phase provides unique information about the essential hypertensive patient. DBP reactivity and drug dose were important determinants of resting blood pressure and mood responsiveness to both pindolol and propranolol. DBP reactivity and perhaps SBP reactivity may be useful measures in the study of central adrenergic and peripheral cardiovascular pathophysiology. PMID- 8226002 TI - Physiologic and psychologic factors related to depression in patients after myocardial infarction: a pilot study. AB - OBJECTIVE: To examine physiologic and psychologic factors contributing to depression in patients one year after myocardial infarction. DESIGN: Convenience, correlational design. SETTING: A university-affiliated cardiology clinic, a health maintenance organization, and a private cardiology group in a southwestern city. SUBJECTS: Fifteen men and six women 10 to 14 months after myocardial infarction. OUTCOME MEASURES: Oxygen saturation, somatic symptoms, and functional status in three areas (social, physical, and emotional) were correlated to depression scores by use of the Zung Self-rating Depression Scale (SDS) and the Symptom Questionnaire (SQ). RESULTS: Spearman rho correlations showed that depression measured with the Zung SDS and SQ depression subscale were inversely related to somatic symptoms, social function, physical function, emotional function, and total function. There was a significant inverse relationship between depression 9as measured by the Zung SDS) and both oxygen saturation and physical function. Although there was an inverse relationship between depression (as measured by the SQ subscale) and both oxygen saturation and physical function, it was not statistically significant. A Mann-Whitney U test for differences showed that mean SDS scores for Hispanic subjects (n = 5) and white subjects (n = 16) differed significantly. Stepwise multiple regression showed that overall functional status accounted for 56% of the variance for depression in the SDS and for 59% in the SQ depression subscale. Oxygen saturation accounted for less than 1% of the variance in both depression scales. CONCLUSIONS: The results of this study indicate that depression is negatively correlated with overall functional status in a select group of patients one year after myocardial infarction. A moderate negative correlation was also found between depression and hypoxia. Hispanic subjects had significant differences in their mean Zung SDS scores as compared with white subjects. PMID- 8226003 TI - ECMO for cardiac support in children. AB - Extracorporeal membrane oxygenation (ECMO), designed to support the lungs in respiratory failure, is less familiar to the nurse who cares for ECMO patients when it is used to support the smaller child or neonate after cardiac surgery. An overview of ECMO, a description of the physiologic response to the ECMO circuit, and a discussion of myocardial function of the patient receiving ECMO are presented. The article focuses on the challenges posed by cardiac ECMO and the nursing care required to meet them. The unique problems addressed include variations in patient cannulation, left ventricular venting, hemostasis issues, the need for hemofiltration, and problems associated with evaluating cardiac function of the patient receiving ECMO. PMID- 8226004 TI - Chloral hydrate side effects in young children: causes and management. AB - OBJECTIVE: To determine the factors related to undesired effects of chloral hydrate in young children undergoing echocardiogram. Undesired effects studied were reaction to chloral hydrate before to sedation (ataxia, excitement), delayed sedation, light sleep during sedation, and behavioral changes after sedation. DESIGN: Descriptive, correlational design. SETTING: Echocardiography laboratory in a pediatric teaching hospital. SUBJECTS: 140 children aged 0 to 36 months who were undergoing diagnostic echocardiography. Severity of cardiac disease ranged from benign murmur to uncorrectable lesion. Thirty children (21%) had cyanotic cardiac disease. Children were sedated with chloral hydrate per routine (mean dose 87 mg/kg) and observed from time of sedation throughout the examination. Data were collected on child's age, food ingested before sedation, transcutaneous oxygen saturation, and daytime nap schedule. OUTCOME MEASURES: Incidence of paradoxical excitement before sedation, length of time until child reached deep sedation, depth of sleep during the examination, and behavioral changes after sedation. RESULTS: Paradoxical excitement before sedation occurred in 25 children (18%). Length of time until child reached deep sedation averaged 25 minutes. Three children never fell asleep. Proximity of sedation to naptime was positively correlated to the speed of sedation. Deep sedation was achieved in 131 children (94%). Depth of sleep during the examination was related to child's age, proximity of sedation to nap time, and recent food ingestions. Older children, who were due for a nap and who had refrained from eating before the examination were most likely to remain soundly asleep throughout the examination. Children with cyanotic defects were not adversely affected by deep sedation with chloral hydrate. Most children experienced motor and affective changes after sedation. CONCLUSIONS: In this sample, chloral hydrate was an effective and safe sedative. Implications for nursing include changes in scheduling practices, limiting food ingestion before sedation, and information to provide parents about chloral hydrate sedation. PMID- 8226005 TI - Readability levels of selected hypercholesterolemia patient education literature. AB - OBJECTIVES: To assess the readability of selected hypercholesterolemia print materials, summarize the limitations of readability formulas, describe how expert judgement can be used to enhance readability determinations of printed materials, and discuss indirect and direct methods for assessing patient literacy levels. DESIGN: Descriptive, retrospective, convenience sample OUTCOME MEASURES: Readability of four AHA/NLHBI hypercholesterolemia patient education pamphlets by use of three readability formulas and the Reading Materials Checklist. RESULTS: The FOG, Fry, and SMOG formulas were used to calculate reading levels by two raters who independently applied the formulas to the same word passages chosen from the beginning, middle, and end of the pamphlets. The mean reading grade levels were 14.4, 15.8, 14, and 14.4, demonstrating that all four pamphlets were written for people with college level reading skills. The limitations of readability formulas such as variations in reading estimates were demonstrated. When expert judgement was applied with the Reading Materials Checklist, all pamphlets were found lacking in the areas of legibility, usability, and motivational appeal. CONCLUSIONS: The results indicate that the pamphlets may not be appropriate for use with most of the adults in the United States who may be candidates for hypercholesterolemia patient education. Because reading formulas are limited in the information they provide, expert judgement regarding readability of print materials should also be applied. Additionally, indirect and direct techniques need to be used to assess the literacy level of the patient population who will be using the printed materials. PMID- 8226006 TI - A comparison of Australian and American registered nurses' use of life-sustaining medical devices in critical care and high-dependency units. AB - OBJECTIVE: To explore how and what Australian and American registered nurses working in critical care and high-dependency units initially learn about the life sustaining medical devices they use as well as the consequences of device use both for patients and staff. DESIGN: Cross-sectional survey with a mailed questionnaire. SETTING: Large midwestern American tertiary care center and two large South Australian tertiary care centers. SUBJECTS: Thirty-two American registered nurses who worked in critical care and 127 Australian registered nurses who worked in critical care, coronary care, and high-dependency units. RESULTS: The most frequently identified initial method of learning both for Australian and American registered nurses was trial and error (taught self). A significantly larger proportion of American nurses initially learned to use a life-sustaining device by watching a video or slide tape, whereas a significantly larger proportion of Australian nurses received instruction in nursing school. At least 90% of Australian and American nurses indicated they initially had learned the same four facts about the device: its purpose, its function, how to respond to the alarms, and how to operate it. A significantly larger proportion of Australian nurses indicated that they had learned about the potential hazards for patients than did their American colleagues. The potential consequences of using any medical device were nurse stress and patient harm. The two reasons most frequently cited by Australian and American nurses for stress were fear of harming the patient and being unsure how to use the device. Of the Australian and American nurses combined, 12.3% indicated they had used a medical device that had harmed a patient. CONCLUSIONS: Australian and American registered nurses were more similar than dissimilar in terms of how and what they learned about the life sustaining devices they use in direct patient care, in experiencing stress when using medical devices, and in having used a device that caused patient harm. Differences in how they learn may be a reflection of marketing and inservice support by manufacturers and basic and highest levels of nurse education. The latter may also account for differences in what Australian and American nurses learned. The results emphasize that nurses must be knowledgeable about the devices they use on behalf of patients. More knowledge about the devices they use in direct patient care might decrease nurse stress. PMID- 8226007 TI - Peripherally inserted central venous catheters in critical care patients. AB - OBJECTIVE: To describe the clinical outcomes and cost for high-risk critical care patients with peripherally inserted central venous catheters (PICCs). DESIGN: A prospective descriptive study. SETTING: A private acute care facility. SUBJECTS: Ninety-seven consecutive PICC insertions in four critical care units per physician order. Five subjects were excluded due to five abortive attempts. Data were analyzed on 92 PICC insertions. Age range was 23 to 92 years. OUTCOME MEASURES: Frequency of insertion complications, postinsertion complications, and cost factors were measured. RESULTS: When subjects were in critical care areas, catheters remained in place a mean of 10 days with a range of one to 50 days. After patient transfer with catheters in place to acute care areas (medical surgical units), convalescent units, and home care, catheters were maintained a mean of 13 days with a range of 1 to 116 days. Catheter days totaled 2069. The total number of catheter days while in critical care were 878, with 1191 catheter days after transfer to other areas. Seventy-one of the original 92 PICC catheters remained in place until treatment was completed or the patient expired. Twenty one were removed for nonelective reasons. Of these patients, only one had significant morbidity related to sepsis. This septic episode occurred in a convalescent unit. Percentage of catheter-related sepsis was 0.48% per 1000 catheter days. Because PICCs are not used to any degree in critical care patients throughout the United States, this study should increase awareness among practitioners that this approach is a safe alternative method to chest or neck insertion for central venous access in high-risk critical care patients. PMID- 8226008 TI - A comparison of pulmonary artery, rectal, and tympanic membrane temperature measurement in the ICU. AB - OBJECTIVE: To compare tympanic membrane temperature with pulmonary artery (PA) and rectal temperature (calibrated glass mercury) to determine consistency among measures. DESIGN: Convenience, within-subject, quasi-experimental. SETTING: Midwestern university-affiliated tertiary medical center. PATIENTS: 128 adult (18 years or older) patients admitted to the surgical intensive care unit with an age range of 18 to 90 years (mean 57 years). OUTCOME MEASURES: Tympanic, PA, and rectal temperature. INTERVENTION: Rectal and tympanic membrane temperatures were measured in 60 patients. PA and tympanic membrane temperatures were measured in 68 patients. RESULTS: Rectal and tympanic membrane temperatures were moderately correlated (r = 0.525). Mean rectal was slightly higher than mean tympanic membrane temperature (mean difference 0.19 degrees C). PA and tympanic membrane temperatures were highly correlated (r = 0.909). Mean tympanic membrane temperature was slightly higher than mean PA temperature (mean difference 0.42 degrees C). For PA temperatures, 57 (84%) of the differences were between 0 and +1 degree Celsius, whereas for rectal temperature, 23 (37%) were between 0 and +1 degree Celsius and 28 (47%) were between 0 and -1 degree Celsius. CONCLUSIONS: Tympanic temperature measurements would be an appropriate substitute for PA temperature if the PA catheter has been removed or the PA thermistor is nonfunctional. In addition, this study suggests that to effectively track temperature in a particular patient, the site for temperature measurement must be consistent. PMID- 8226009 TI - Do age and gender influence outcome from mechanical ventilation? AB - OBJECTIVES: To define the role of age and gender on outcome from mechanical ventilation. DESIGN: Prospective analysis of consecutive patients. SETTING: A military-referral medical center. PATIENTS: A total of 240 consecutive patients requiring 246 episodes of mechanical ventilation. MEASUREMENTS: Demographic, clinical, and outcome data. RESULTS: Univariate statistical techniques comparing 13 variables between survivors and nonsurvivors of an episode of mechanical ventilation indicated that the following eight variables were significantly different (p < 0.05) between these two groups: age, length of intensive care unit stay, duration of MV, Acute Physiology and Chronic Health Evaluation (APACHE) II score, Age Removed APACHE II score, Organ System Failure Index (OSFI), patient diagnostic category, and presence of malignancy. Patient sex demonstrated a trend towards significance (p = 0.051). These nine variables were entered into a stepwise logistic regression analysis to develop a model to predict outcome from mechanical ventilation. This model showed that only the OSFI (p < 0.001) and the APACHE II score (p = 0.0281) independently predicted outcome from mechanical ventilation. Similar analyses were performed to identify outcome predictors after segregating the patients according to age (less than 70 years and equal to or greater than 70 years) and according to sex. In the first of these subsequent analyses the OSFI was found to be the sole independent predictor of outcome for both older and younger subgroups. In the second analysis the OSFI, length of hospital stay, and duration of mechanical ventilation predicted outcome for male patients while the OSFI alone independently predicted outcome for female patients. Fifty-two survivors of mechanical ventilation over the age of 70 years were observed after hospital discharge (average follow-up time 2.75 +/- 1.36 months) to assess their need for institutional care after hospital discharge. Five of these patients (9.6%) required long-term care in a nursing home or rehabilitation hospital until their deaths (four patients) or discharge to home (one patient). With a stepwise logistic regression analysis, only the intensive care unit length of stay was found to be significant independent predictor for this outcome. CONCLUSIONS: These results suggest that the occurrence of derangements in organ function as described by the OSFI best predicts outcome from mechanical ventilation. Age and sex do not significantly contribute to patient outcome independent of organ system dysfunction. PMID- 8226010 TI - Hepatitis Part II. Viral hepatitis B, C, and D. PMID- 8226011 TI - In vitro survival of skin flora in heparin locks and needleless valve infusion devices. AB - OBJECTIVE: To determine the extent to which two types of infusion devices, the heparin lock and a needleless valve device, allowed the persistence of inoculated microorganisms. DESIGN: Experiment. SETTING: Clinical microbiology laboratory. OUTCOME MEASURES: Bacterial counts in infusion devices. INTERVENTION: Five of each type of device were inoculated with approximately 10(5) colony-forming units of a strain of Staphylococcus epidermidis and Enterobacter aerogenes and tested for growth at seven time intervals, from 10 minutes to 72 hours after inoculation. RESULTS: Both strains of bacteria were present in each device at every time interval tested, including 72 hours after inoculation. CONCLUSIONS: Bacteria introduced into a heparin lock or valve device may be isolated for prolonged periods of time. This suggests that if such devices are contaminated during use, they may be a potential source of infection. The risk of infection should be one major consideration in risk and benefit deliberations when choosing new products. PMID- 8226012 TI - Liver laceration after cardiopulmonary resuscitation: a case report. PMID- 8226013 TI - Spotlight article: a preliminary study of diltiazem in the prevention of coronary artery disease in heart-transplant recipients. (Schroeder JS, Gao SZ, Alderman EL, et al. N Engl J Med 1992; 328:164-70.). PMID- 8226014 TI - Technical aspects of pulmonary artery catheter measurement. PMID- 8226015 TI - Sharps disposal containers. PMID- 8226016 TI - Standards: what's new in NFPA 99-1993? AB - Another revision cycle for the National Fire Protection Association's (NFPA) Standard for Health Care Facilities (ANSI/NFPA 99, An American National Standard, February 12, 1993) was recently completed. The 1993 edition incorporates numerous changes. Below, we present a summary of the changes that relate to electrical systems and electrical safety. Extensive changes have also been made to the chapters dealing with medical gas systems; we will be commenting on these revisions in an upcoming Health Devices article on medical gas systems, and we will discuss other changes in future issues as the need arises. PMID- 8226017 TI - Guidance article: cellular telephones and radio transmitters--interference with clinical equipment. AB - It has long been recognized that devices emitting radio frequencies can interfere with the operation of clinical and other electronic equipment. A number of Health Devices System members have called us in recent months to ask about the risks of using cellular telephones in hospitals, and several others have also raised similar questions in the Healthcare Technology conference area of ECRINet, our online communication service. While some of those making inquiries have been alarmed by the publicity surrounding the possible health hazards of using cellular telephones--a specter that is not supported by currently available evidence or theoretical considerations--the focus of concern is generally whether such telephones can affect the operation of patient care devices. Evidence indicates that operating cellular telephones and radio transmitters near certain clinical equipment presents the risk of interrupting patient care, although much work remains to be done to clearly define that risk. Nevertheless, we recognize the benefit of bringing to the attention of our readers some of the experiences that are being reported. We hope that the following questions and answers will increase awareness of the possible risks of using such equipment, as well as promote the exchange of information to permit a more rapid assessment of the degree of risk and the steps necessary to avert problems. PMID- 8226018 TI - Electrical safety questions and answers. PMID- 8226019 TI - Burns and fires from electrosurgical active electrodes. PMID- 8226020 TI - Inadvertent dislodging of loading units from United States surgical autosuture ENDO GIA 30 surgical staplers. PMID- 8226021 TI - Misleading battery-charging indicator on Marquette TRAM Series 300 and 360 defibrillators. PMID- 8226022 TI - Patient-accessible rotating-gantry components in GE HiSpeed Advantage CT scanners. PMID- 8226023 TI - Restricted draw in Schrader-type vacuum inlets. PMID- 8226024 TI - Home care for the person with cancer. AB - Nearly 90% of all cancer care is delivered in outpatient settings (including home care). Home care for the person with cancer combines the routine objectives of a comprehensive home healthcare program with services especially designed for individuals with cancer. A key component is the involvement of and concern for family members or other designated caregivers. This article reviews interventions most often incorporated into home care plans, including parenteral and enteral nutrition; pain management; antimicrobial, antiemetic, immunosuppressive, and blood component therapies; and administration of biologic response modifiers. Descriptions of cancer-related resources useful to the home care nurse are included. PMID- 8226025 TI - Advance directives: a program design for home healthcare. AB - Home care agencies receiving Medicare and Medicaid reimbursement are now required to adhere to the regulations of the Patient Self-Determination Act of 1990. These regulations require all agencies to inform clients older than 18 years of their right to plan for their care in advance, and to provide clients with the agency's written policy respecting this right. It also requires that agencies educate their staff about the new provisions. This report presents a program designed to orient all new and continuing home care staff members with the requirements to ensure compliance with this important directive. PMID- 8226026 TI - Cancer pain management. AB - Today we have the necessary cancer pain-relieving agents. These medications can safely be administered orally and in the patient's home. Sufficient analgesia offers the patient the opportunity for a dignified and comfortable death. The family is relieved from the anger, despair, and sense of helplessness that comes from watching a loved one die in uncontrolled pain. As nurses in home healthcare, we can impact the statement that 25% of all cancer patients die without pain relief. It is our mission to integrate all that is known about pain management into a holistic practice that will provide the patient with the opportunity for a dignified and comfortable transition from this life. PMID- 8226027 TI - Social support in home health nurses. AB - The decision of a home health nurse to remain employed at an agency is certainly influenced by a number of factors, one being how much support or encouragement that nurse is receiving from colleagues. The home health nurse does interact with peers at the office and over the telephone, but such an act as calling down the hall to another registered nurse to help with a client is not possible. PMID- 8226028 TI - Chris King: extraordinary nurse, unforgettable human being. AB - Chris King, a home health nurse, an employee, and my friend, was one of the strongest women I have ever known. Chris died in May 1990; she was only 40 years old. This article is dedicated to her memory. PMID- 8226030 TI - Priorities. PMID- 8226029 TI - Chemotherapy: how safe for the caregiver? AB - Antineoplastic drugs are routinely administered by nurses in the home setting. The potential for nurse occupational exposure to these drugs is increased by an inadequate understanding of the risks inherent in administering these agents. This article addresses issues related to safe administration of these drugs in a home environment. PMID- 8226031 TI - Getting personal. PMID- 8226032 TI - The peer review organization process revisited. AB - The outcomes of patient care are of increasing importance in the 1990s from a quality of patient care perspective. Most of the review processes that HHAs are subjected to, including the Medicare certification survey, the accreditation process by the Joint Commission on the Accreditation of Healthcare Organizations and the Community Health Accreditation Program, are focused toward patient outcomes. Also included is the PRO process that addresses the quality of care issue when home care was provided as one of the intervening care services if a patient is readmitted to an acute care facility within 31 days of discharge, HHA standards as well as professional nursing standards. There is no question that the care rendered and the complete and accurate documentation by home healthcare nurses contribute to the minimal number of confirmed quality of care problems that have been identified in HHAs. We would appreciate findings reported in other states. Please share published information on the PRO activities in your state in care of Home Healthcare Nurse. We look forward to hearing from you. PMID- 8226033 TI - Home health/public health/visiting nurse returning to our past: a comparison of public health nursing at the turn of the century. AB - The basic ingredients that made early home care a cornerstone in American healthcare are still the foundations for home health/visiting nurse/public health in the 1990s. Once overshadowed by the centralization of patient care in the hospital, home care has stepped into the place of cooperative healthcare with the hospital. This article highlights that cooperation and those basic ingredients. PMID- 8226034 TI - [Surgery for bleeding peptic ulcer: short- and long-term results]. AB - Thirty-one patients operated on for bleeding peptic ulcer were reviewed. The basic concept was to make an early decision to operate and proceed as soon as the patient was haemodynamically stable. In addition to haemostasis, a definitive operation was performed. The procedure was a proximal gastric vagotomy (PGV) for duodenal ulcers (DU), combined with an antrectomy for pre-pyloric ulcers, and either a PGV with ulcer excision or a Billroth I for gastric (GU) or combined (GU + DU) ulcers. Twenty-four patients (77%) were operated on within the first 24 hours. Nine patients could not be operated according to the basic protocol because of anatomical reason, additional ulcer complication or severe co-existing systemic disease. During the hospital stay, 2 deaths (6%) occurred and 4 patients (13%) rebled postoperatively, all of them were reoperated. During a mean follow up of 44 months, 12 deaths unrelated to peptic disease and one recurrent bleeding occurred. PGV for DU could be used in 70% of cases without any hospital mortality; one patient rebled after the operation and another during the long term follow-up. These results support the views that early surgery has a low hospital mortality and that PGV gives good results when performed as an emergency procedure. PMID- 8226035 TI - [Perioperative morbidity and mortality in colon resection for colon cancer]. AB - The operative risk of colon resections was evaluated by a retrospective analysis of 231 according patients who were operated on between 1984 and 1988. Mean age of the patients was 70 years with a range from 37 to 91 years. Colonic resection consisted of ileocecal resection in 3 cases, right hemicolectomy in 144 cases, segmental resection of transverse colon in 10 cases, left hemicolectomy in 22 cases, resection of sigmoid colon in 77 cases and 5 times a subtotal colectomy was performed. In two patients (0.9%) an anastomotic leak occurred. Three patients were reoperated on: one due to an anastomotic disruption, two others due to a mechanical small bowel obstruction. Two patients (0.9%) died due to systemic complications without any evidence of anastomotic or wound problems. Thus a low morbidity and mortality of colonic resection is documented in our study. Factors contributing to these results are a standardized bowel preparation, perioperative antibiotics and modern anaesthetic techniques. PMID- 8226036 TI - [The value of conservative therapy, surgical treatment and interventional radiology in perforations and ruptures of the esophagus]. AB - We have reviewed all the records of 23 patients hospitalized in our institution from January 1981 till December 1991 and presenting a confirmed diagnosis of esophageal perforation. We have studied the aetiology, the localization, morbidity and mortality and we discuss the therapeutic management. 9 patients presented a cervical perforation, 13 patients a thoracic perforation and 1 patient an abdominal perforation. Among the patients with cervical perforation 2 patients had a local revision with drainage, 3 patients a primary suture and 4 patients were treated conservatively. No complication was found in this group. The patients with thoracic perforation have been treated as follow: 3 conservatively, 8 with thoracotomy, primary suture +/- patch, drainage, 2 patients with thoracotomy and drainage alone. All complications happened in this group: 2 gastro-intestinal bleeding, 2 ARDS, 3 mediastinitis, 1 pneumonia. 2 patients in a very poor general condition died, one with a metastatic breast carcinoma, the other after a CVI with a massive gastro-intestinal bleeding. The cervical perforations have an excellent prognosis and can be treated conservatively if they are asymptomatic and do not display a pleural lesion. The thoracic perforations can be treated surgically if they are diagnosed early before septic complications. If not, they will be better treated conservatively with drainage. The intraabdominal perforations have to be treated as every intraabdominal perforation. In this case, we perform a primary suture completed with fundoplication. PMID- 8226037 TI - [Retroperitoneal sarcoma. Therapy and course]. AB - The incidence of retroperitoneal soft tissue sarcomas is with 0.1 to 0.2 percent very low. The tumors most often present as asymptomatic masses until the lesions are quite large, compared with the anatomic part, resulting in compression of nerves, vessels, bowel etc. The recommended management is the aggressive radical tumor excision including the pseudocapsule followed by a consequent clinical, ultrasonography and CT-scan follow-up. Tumor recurrences should be attempted in the same manner as primary tumors. PMID- 8226038 TI - [Surgical-radiologic interventional treatment in liver abscess with Streptococcus anginosus Milleri]. AB - Pyogenic liver abscess is a relatively rare disease, often concerning elderly patients in bad general condition and with underlying diseases. Out of a retrospective study (1984-1991) of 44 patients (f 43%, m 57%) with pyogenic liver abscesses and 12 patients (f 17%, m 83%) with amebic liver abscesses we present 5 patients with multifocal lesions due to Streptococcus anginosus Milleri--a special group concerning therapy and course of disease. The therapy was high-dose i.v. antibiotic therapy in all 5 cases and additional percutaneous drainage with pig-tail catheters--inserted under CT guidance--in 4 cases (1-5 catheters, time of drainage 6-63 days). The duration of hospitalisation was 15-73 days (median 45), intravenous antibiotic treatment lasted 4-40 days. After discharge oral antibiotics were continued for 14-42 days. PMID- 8226039 TI - [Solid and cystic pancreatic tumor]. AB - The solid and cystic tumor of the pancreas (also called pancreatic cystic tumor of the pancreas) is a very rare tumor of the pancreas, which we meet mainly in younger women. The tumor was described for the first time in 1981 and since that time there were published more than 150 cases. It is a matter of a potentially malign tumor with low malign characteristics. By radical exstirpation most of the patients can be healed definitely. Long-term controls are yet necessary although recidives are rare (4-7%), but are described up to 10 years after the first treatment. Also singular cases with synchronous metastasis of the liver are mentioned. Except of abdominal volume increase the tumor gives little trouble. The diagnosis is placed with ultrasonography resp. with a computer-tomogram and ensured by needle cytology. The therapy consists in the radical exstirpation with resection of the left pancreas in case of location on the left side, with Whipple operation in case of localisation in the head of the pancreas. PMID- 8226040 TI - [Lymphoepithelial cyst of the pancreas]. AB - In a routine check-up a cystic tumour of the head of the pancreas was found in a asymptomatic 59-year-old man. As the investigation with several radiological means was not conclusive and there was found an increased value for CEA in the cysts liquid a explorative laparotomy was performed. Already the intraoperatively performed histology confirmed the diagnosis of a benign lymphoepithelial cyst of the pancreas. After local resection the postoperative course was uneventful and the patient is today, one year later, disease-free. Lymphoepithelial tumours are well known in the parotis whereas in the pancreas only 5 cases have been reported. For positivity for CA 19-9 marker we found some evidence for the hypothesis that these kind of cysts could derive from metaplastic tissue in obstructed pancreatic ducts. PMID- 8226041 TI - [Association of a giant Meckel's diverticulum and Crohn disease of the neighboring intestinal segments]. AB - We report the case of a 51-year old patient presenting a giant Meckel's diverticulum associated with Crohn's disease in the proximal and the distal segments. The voluminous diverticulum had already been demonstrated by radiographies at the age of 28 years. Late symptoms were signs of partial obstruction and malabsorption. PMID- 8226042 TI - [Misinterpretations in arteriography of the lower extremities]. AB - Arteriography is still the most important diagnostic tool in the assessment of chronic lower limb ischemia. Unfortunately, demonstration of distal vessels is often poor. Inadequate arteriography makes decisions on the feasibility and the site of reconstruction impossible. Methods to improve the quality of arteriograms include correct positioning of the legs in internal rotation, long exposure times, additional lateral views and the capacity of digital subtraction. The importance of positioning the legs in internal rotation is illustrated by several examples. PMID- 8226043 TI - [How often does an aneurysm of the popliteal artery rupture?]. AB - Rupture of a popliteal artery aneurysm is an unusual complication and did occur six times in our series of 252 popliteal artery aneurysms that underwent operative repair from January 1965 to December 1991. Rupture while equally serious as thrombosis of the popliteal artery aneurysm of distal embolization from the aneurysm, is reported as an exceedingly unusual complication. Reported incidence of rupture is less than 2 to 4 percent in most large series and was 2.4 percent in the present series. PMID- 8226044 TI - [Value and use of pedal reconstruction in an extremity at risk for amputation]. AB - In patients with critical limb ischemia and patent pedal arteries as a single outflow source the surgeon has to decide between primary amputation and reconstruction. From 1989 to 1991 we performed 80 infrainguinal reconstructions, 13 of them to pedal arteries. In all cases we used autogenous vein material from different sources, if possible as an in situ conduit. The perioperative mortality was 0%. After 24 months the limb salvage was 92%. The secondary cumulative patency rate was 92% after 12, 74% after 24 months. In a selected group of patients we prefer arterial reconstruction to pedal arteries instead of a primary amputation. PMID- 8226045 TI - [Dacron prosthesis dilatation. Case report and review of the literature]. AB - Twelve years after implantation of a subclavian-femoral Dacron prosthesis the graft had to be replaced because of diffuse aneurysmatic dilatation. There was no infection documented. Dilatation of knitted Dacron prostheses under arterial pressure is a normal event but rarely causes graft failure due to aneurysm formation. The following problems can promote graft dilatation: fabrication defects, multiple sterilization, false storage, damage during the operation, infection or/and hematoma, host vs graft reaction, constant mechanical stress due to anatomical facts, material fatigue and gliding of fibers and chemical reactions because of bioinstability. We recommend to use rather small sized prostheses in the aorto-iliac system. Some authors recommend long-term follow-up of all patients with artificial grafts. PMID- 8226046 TI - [Aorto-duodenal fistula--a late complication after infrarenal aortic replacement]. AB - Peripheral atherosclerotic vascular disease is associated in up to 12% with an abdominal aortic aneurysm (AAA). If operation is required AAA is treated in 90% by infrarenal dacron graft implantation. As late complication secondary aortoduodenal fistula is diagnosed in 0.1-4.0%. Mortality in untreated patients is 100%, perioperative mortality depending on operative technique is 17-77%. We present 2 case reports and diagnostic, operative and prophylactic methods. PMID- 8226047 TI - [Thrombosis of the ascending aorta--a rare cause for recurrent arterial embolism]. AB - A 41-year-old man suffered 3 episodes of transitory left leg ischemia before he was admitted with right brachial artery embolism. The diagnostic evaluation (i/v DSA, CT Scan) revealed a mass in the ascending aorta which was removed under hypothermic cardiocirculatory arrest. Histologically the mass was shown to be a thrombus adjacent to a arteriosclerotic plaque. PMID- 8226048 TI - [Therapy of pulmonary arteriovenous aneurysm]. AB - Connections of branches of the pulmonary artery to branches of the pulmonary vein resulting in aneurysmatic dilatation are defined as pulmonary arterio-venous aneurysm (pava). The spontaneous course shows a morbidity of 26% and a mortality of 11% within a six-year observation period following diagnosis. If there is an arterial branch with small diameter (< 15 mm) we recommend the embolisation with the catheter; this procedure is less stressing for the patient and does not involve the loss of pulmonary tissue. If the arterial branch has a big diameter, if a complex pava or a pulmonary abscess is present a conventional surgical resection should be envisaged. PMID- 8226049 TI - [Etiology and therapy of pneumothorax in AIDS patients]. AB - Pneumothoraces in patients with AIDS are a rare, acute and severe complication. Between January 1989 and December 1991 3 patients with spontaneous pneumothorax were operated after a 2-week treatment with chest tube drainage only had failed. The average follow-up time after the operation was 7 months. There was no recurrence. Within the same 3 years 3 patients were treated in the Clinic for Internal Medicine by means of tube drainage only. Thus, only one case was successful. The average survival after receiving the tube was 3 weeks. Their death was not related on their pneumothorax. The mean age of the 6 patients was 34 years (23-49). In 5 patients the pneumothorax was associated with Pneumocystis carinii-pneumonia and in 3 of them also with prophylactic pentamidine aerosol therapy. A minimal invasive operation of operable AIDS-patients with pneumothorax shows good results, shortens the time of therapy and is possible despite the bad prognosis of the disease. PMID- 8226050 TI - [Early results of surgical revascularization of the subclavian artery]. AB - In the last 14 years (1978 to 1991) in the Clinic of Cardiovascular Surgery, University Hospital Zurich, 72 patients were operated due to obstruction of the subclavian artery. The left side was affected in 46/72 (63.9%), the right side in 21/72 (29.2%), both sides in 5/72 (6.9%) patients. Only thrombendarterectomy has been done in 48/72 (66.7%), followed by carotis-subclavian-bypass (9/72, 12.5%). Furthermore the following techniques were applied: endarterectomy and patch in 6/72 (8.3%), aorta-subclavian-bypass in 5/72 (6.9%), bypass between proximal and distal subclavian artery in 2/72 (2.8%), resection and graft-interposition in 1/72 (1.4%) cases. As well as operation-related reactions of soft tissues (hematoma, swelling) the following complications have been recognized: re stenosis with embolus or intima-flap in 8/72 (11.1%) cases, 3-times persistent, paralysis of the vocal cord in 3/72 (4.2%) cases, phrenic nerve paralysis in 2/72 (2.8%), in two separate cases Horner's syndrome and pneumothorax. The oldest patient (81 years) died 10 days after operation due to hemorrhage, in spite of 3 re-thoracotomies. Additional complications arose independently of the subclavian operation (atrial flatter, heart-infarction, lungoedema, exitus following extensive heart-operation with valve-replacement and aorto-coronary bypass). PMID- 8226051 TI - [Paraplegia, a catastrophic complication of interventions on the infrarenal aorta]. AB - Damage to the spinal cord in course of the treatment of diseases of the infrarenal aorta is a rare but calamitous complication. The reported incidence is about 0.2%. The neurological loss is usually complete flaccid paraplegia with high mortality and rare full or partial recovery. Between 1980 and 1991, 1070 reconstructive procedures of the infrarenal aorta were performed: 821 due to aneurysm (316 elective procedures [mortality 1.6%] and 505 emergency procedures [mortality 24.5%]) and 249 due to aorto-iliac occlusive disease. Damage to the spinal cord occurred in 2 patients (2/1070, 0.19%). One patient had incomplete paraparesis following repair of an unruptured abdominal aortic aneurysm with gradual return of all neurological symptoms to normal. The second patient developed complete paraplegia following repair of a ruptured infrarenal aneurysm. There war no recovery of the symptoms. The patient died from septicaemia 4 months later. PMID- 8226052 TI - [Results of surgical revascularization of the kidney]. AB - There are many different etiologies of renal artery disease (atherosclerosis, aneurysm, dissection, arteriovenous fistula, embolism, fibromuscular dysplasia) and also a lot of different therapies (conservative treatment, percutaneous transluminal angioplasty [PTA], endarterectomy, bypass grafting, patch plasty, nephrectomy). Recently conservative treatment and PTA have significantly improved. Patients who are referred to surgery today are of older age with severe and often bilateral disease of the renal arteries. Additional manifestations of general atherosclerosis like coronary artery disease, aortic aneurysm, peripheral occlusive vessel disease and cerebral vascular insufficiency are often present as well. The main goal of all forms of treatment is the preservation of general renal function. With our retrospective study the results after surgical revascularisation of kidneys are evaluated over a short period of time. Only graft revascularisations are included in the study and we were mainly interested in renal function and blood pressure. PMID- 8226053 TI - [Case report of spontaneous splenic rupture in splenoma]. AB - We report one case of splenoma, a rare benign tumor of the spleen. Initial symptoms were repeated attacks of tail pancreatitis, that led to splenectomy for spontaneous splenic rupture. Splenomas may be asymptomatic or cause haematologic symptoms. Recently their association with neoplastic and haematologic disorders has been emphasized. Therefore such diseases must be ruled out when a splenoma is diagnosed. PMID- 8226054 TI - [Traumatic splenic rupture: when surgical treatment? When conservative treatment? A retrospective study based on 56 cases of traumatic splenic ruptures]. AB - In a retrospective study of 56 cases of traumatic spleen injury we could show that neither ultrasound nor CT-scan influence directly the treatment modality (operative or conservative). Only clinical parameters are of importance. PMID- 8226055 TI - [Computerized tomography follow-up in the acute phase after craniocerebral trauma]. AB - Because of methodological reasons, within the first day after a head trauma, CT scan often does not demonstrate all the pathological consequences of brain injury. Additionally, secondary sequelae are frequent events. In our series of 129 patients the findings on subsequent CT scans done during the posttraumatic course were worse compared to the initial CT in 45% of patients. This deterioration on the CT scan was seen irrespective of the initial GCS score and the initial diagnosis of circumscribed or diffuse injury. The event observed most often was bleeding into a preexisting contusion or even bleeding into a region which had looked normal on the first CT scan. In the vast majority of cases, this happened within the first 2 weeks after the initial trauma. We conclude that an initially pathological CT should be repeated early in the posttraumatic period even in patients with mild to moderate head injury. In particular, we recommend that patients having sustained diffuse brain injuries should be transferred to neurotraumatology units for specialised management, even when there may not be immediately obvious indications for surgical intervention. PMID- 8226056 TI - [Quality of emergency admission (resuscitation, REA) and first aid in multiple trauma]. AB - Undue delay between hospital admission and the beginning or urgent operative procedures is considered as a major mortality risk for polytraumatized patients in any trauma center. As part of a quality control study at our institution (Kantonsspital, University of Basel), the time spent for early resuscitation and diagnostic procedures was therefore prospectively recorded in 20 patients (mean age 38 years) with a mean ISS of 26.9 (range: 13 to 43). Time spent in the resuscitation room averaged 31.4 min (range: 10 to 50 min). Conventional radiographic diagnostic procedures took 34.7 more min (range: 20 to 60 min). An additional CT scan was performed in 15 patients requiring 19.5 min per region (head/thorax/abdomen/spine). Four patients underwent angiography necessitating 28 more min (mean). Time elapsed between admission and arrival of the patient in the OR or the ICU respectively accounted for an average of 89 min (range 22 to 200 min). For comparable injury severities this interval was shorter during the day than during the night (77 and 103 min respectively). Diagnoses established during this period were both accurate and comprehensive, as detectable from the low rate of missed diagnosis (three minor fractures). Although our results match favorably with figures reported in the literature we feel that further improvements could be achieved by performing the conventional radiographic procedures simultaneously with the early resuscitation in the resuscitation room. At present time, for reasons of X-ray protection, this is not possible in our institution. PMID- 8226057 TI - [Reconstruction of the lower extremity using partially destroyed or amputated parts]. AB - We are often confronted with heavy destructions of the lower extremities, where there is no possibility to conserve the limb, because of the general or/and local conditions. In this situation it's wise to preserve vital rest structures that can be used for primary or secondary reconstructions of the injured limb. There are different possibilities, some of them are presented in the cases described in this article. PMID- 8226058 TI - [Blastoma of the lung--a rare malignancy. Case report]. AB - Pulmonary blastoma--so called because of its histological resemblance to fetal lung--is a rare malignant neoplasm, which tends to relapse locally and metastasize mainly in the first years, has a poor 5-year prognosis, and shows a potential radio-chemosensitivity. We report our findings and therapeutic approach in a case of a 64-year-old male. PMID- 8226059 TI - [Prospective surveillance of nosocomial infections in a traumatology and orthopedics service]. AB - From May 1989 to February 1990, 1164 patients were followed in the Service of Orthopaedic Surgery and Traumatology at the CHUV, for the development of nosocomial infections (N.I.), in particular postoperative wound infections. Among them, 439 patients were treated conservatively and 725 underwent 833 operations. The global incidence of N.I. was 7.7/100 admissions. Urinary tract infections represented 57% of all N.I., postoperative wound infections represented 21% (with 12% superficial and 9% deep infections), pneumonias represented 20% (5 deaths), bacteriemias represented 2% (1 death) of the cases. The rate of infections was 1.1% for clean surgical interventions (n = 549), 0% for clean-contaminated operations (n = 87), 7.5% for contaminated operations (n = 40), 22.8% for dirty operations and 1.6% for unclassified operations (n = 122). Among the 19 postoperative wound infections, 6 were diagnosed after the patients were discharged. In conclusion, postoperative wound infections constituted only 20% of all N.I. observed in the Service of Orthopaedic Surgery and Traumatology and the rates of infection according to the types of interventions were low. On the other hand, 80% of the N.I. were observed at sites other then the wounds and were associated with a 0.5% mortality rate. PMID- 8226060 TI - [Distal humerus fractures in patients over 75 years of age. Long-term results of osteosynthesis]. AB - The aim of this retrospective analysis was to evaluate the adequacy of internal fixation of distal humerus fractures in patients over 75 years of age. 49 patients were evaluated. The mean age was 80 (75-93) years. The fractures were classified as 28 C-, 13 B- and 8 A-types according to the AO-system. Primary stable fixation was followed by early assisted mobilisation. The mean average follow-up time was 18 months. Excellent and good functional results were observed in 85%. 66% of all patients have no pain. There were 6 sensible ulnar nerve lesions, one deep wound infection, one pseudarthrosis of the distal humerus and one non union of the olecranon osteotomy. We conclude from our results that open reduction and internal fixation is indicated also for patients over 75 years with distal intraarticular humeral fractures. PMID- 8226061 TI - [Monteggia injury in childhood: a frequent cause of chronic dislocation of the head of the radius]. AB - Two cases of chronic dislocation of the radial head after Monteggia lesions in children are described. The late reconstruction with open reduction of the radial head combined with osteotomy of the ulna is mentioned as the therapy of choice before skeletal maturity, because the pathophysiology of the lesion is best considered. The operative procedure in detail is given. Finally, alternative procedures are presented and discussed. PMID- 8226062 TI - [Experience with the AO universal femoral intramedullary nail for management of femur shaft fractures]. AB - 57 fractures of the femoral shaft, treated with an AO femoral interlocking nail, have been analysed in a retrospective study. 40 nails have been implanted primarily, 17 as secondary procedures following different operations. In the group of primary nailing 35/40 reached a good or very good result, 8 of them however, following secondary operations. In the group of the secondary nailing, 9/17 patients had a good or a very good result. 3 pseudarthroses persisted. The AO femoral interlocking nail therefore appears to be an adequate implant for the treatment of femoral shaft fractures, but a precise implantation technique is a prerequisite for the successful outcome. PMID- 8226063 TI - [Load stable, minimal fibula osteosynthesis in Weber type B malleolar fracture by hemicerclage]. AB - Simple lateral malleolar fractures type B according to Weber can be fixed with a hemicerclage alone. Technique and results at sixteen own cases are shown. At this indication the hemicerclage is regarded as a good alternative to common methods of osteosynthesis. Advantages are simplicity, minimal damage of soft tissue and bone, high interfragmentary compression and allowed full weight bearing. PMID- 8226064 TI - [Management of lateral type B malleolar fracture with penetrating wires, cerclage and hemicerclage]. AB - In this retrospective study 52 cases of malleolar fractures type B, that were treated with Kirschner wires, cerclages and hemicerclages, between 1981 and 1991 at the Mannedorf Hospital, were compiled and completed with the corresponding synopsis. The results show that the osteosynthesis technique, in certain cases, is absolutely justifiable and represents a reliable alternative to tensile screw osteosynthesis. This procedure of osteosynthesis is a good method in comminuted fractures as well as osteoporotic bone, in which the screws grip badly. A minimum of osteosynthesis material is implanted in precarious soft tissue conditions. As in the tensile screw osteosynthesis, the removal of the material can be done ambulatory under local anaesthetic with stab incision. The functional after treatment is not recommended. We suggested a postoperative plaster of Paris treatment for 6 weeks (2 weeks without and 4 weeks with strain). As the postoperative course shows comparable results, osteosynthesis of the lateral malleolar fracture type B with Kirschner wires, cerclages and hemicerclages represents a good variation to plate osteosynthesis and tensile screw osteosynthesis. PMID- 8226065 TI - [Metatarsus primus double osteotomy, a logical and non-disabling surgical technique for treatment of hallux valgus]. AB - The procedure known as the Metatarsus Primus Double Osteotomy ("L'osteotomie metatarsienne bipolaire" of Schnepp-Carret) for the treatment of the hallux valgus is a logical, efficient and non-mutilating procedure. The technique consists in reducing the hallux valgus deformity by a subcapital bone wedge resection of the metatarsus primus. The bone wedge is then pinched into a second proximal osteotomy of the metatarsus primus, correcting the varus deformity in a valgus direction. This procedure is little known and is not even mentioned in the classic orthopedic literature of German and English language. PMID- 8226066 TI - [Giant cell tumor--an unusual cause of pathologic fracture of the femoral neck]. AB - The uncommon giant cell tumors of bone are generally benign but locally aggressive with tendency for local recurrence. They are located in the epimetaphysis of the long bones, predominantly in the distal femur, proximal tibia and distal radius; the location in the proximal femur is very rare. Because of their location and the high local recurrence, therapy, mainly of the rare pathological fractures, is difficult and controversial. We are presenting a well documented case with joint salvage and good outcome. PMID- 8226067 TI - [Indications, technique and interpretation of arterial Doppler ultrasound]. AB - Doppler sonography is one of the most important diagnostic tools for angiologists and vascular surgeons, and also for general practitioners with an interest in vascular disease. It can be carried out easily and at low cost and at the same time provides reproducible, quantitative data on with further diagnostic and therapeutic decisions can be based. First, systolic arterial pressure in the anterior and posterior tibial and in the peroneal arteries are measured, with the Doppler probe placed at ankle level. A cuff is wrapped around the lower leg and inflated until the Doppler signal disappears and then deflated. The highest value measured in each leg is termed "ankle pressure". Division of the latter by systolic brachial pressure results in the so-called "ankle-brachial-index" or "ABI". Ankle pressure and ABI correlate well with clinical findings. In normal individuals, the ABI is greater than 1. In claudicators, it ranges between 0.3 and 0.9, in patients with resting pain between 0.1 and 0.5 and with ischemic tissue loss between 0.0 and 0.2. After angioplastic or surgical revascularisation procedures, a fall of the ABI by 0.15 or more is an indication of relevant hemodynamic deterioration and therefore calls for further investigation by arteriography or colour duplex sonography. PMID- 8226068 TI - [Technique for endoscopic vein preparation in infra-inguinal in situ bypass]. AB - In situ saphenous vein bypass for infrainguinal reconstructions achieves excellent results, especially for distal anastomoses. The achilles heel of the method is the complete and atraumatic disruption of the valves and the location of significant tributaries of the saphenous vein. An intraoperative quality control is necessary to mend early patency rates. Several control instruments are described: intraoperative arteriography, duplex- or Doppler sonography and angioscopy. We describe the technique of a semiclosed angioscopy guided technique with a novel flushing valvulotome. We use an angiofiberscope with 2.2 or 1.4 mm outer diameter, being introduced through the proximal exposed end of the saphenous vein. Through the distal end or a side branch a modified Mill's valvulotome with a flushing channel (flexible or non flexible) is moved upwards to the angioscope. Every valve cusp is incised under direct vision in a retrograd direction. Simultaneously relevant tributaries are angioscopically located and ligated through small skin incisions. To use the potential of angioscopy we have developed the occluder valvulotome, making endoluminal embolization of tributaries of the saphenous vein possible. Details of these techniques are described. PMID- 8226069 TI - [A new self-repositioning technique for fresh, anterior-lower shoulder dislocation]. AB - In case of a shoulder dislocation there is always a question of a safe and atraumatic reduction technique if possible without analgetics or even general anesthesia. The dislocated humeral head is reduced anatomically with the help of the patient's own body weight under muscular relaxation without injuring manipulations. Even with displaced fractures of the greater tuberosity or an impression fracture of the humeral head successful reductions were obtained. We have been using this method for over three years with a primary success rate of over 60%. PMID- 8226070 TI - [Surgical technique in ski thumb injury]. AB - The skier's-thumb, an instability of the ulnar collateral ligament (UCL) of the first metacarpo-phalangeal joint is one of the most common hand injuries in alpine skiing. In over 60% of our cases the ligament is ruptured, mostly near or at its distal insertion at the base of the first metaphalanx. Also a bony avulsion of the UCL at its distal insertion is not uncommon. A significant instability or a bony avulsion of the UCL with dislocation of the fragments is an indication for open reconstruction and stabilization. Good experience has been gained with a fish hook pull-out-wire technique. We describe the approach and the reconstruction technique. PMID- 8226071 TI - [Multiple retroperitoneal and mediastinal lymph cysts in primary ectatic lymph vessels]. AB - A unique case of a large retroperitoneal and multiple mediastinal lymphcysts is presented. Cystic abdominal tumors of lymphatic origin are very rare. Usually single cases or small numbers have been reported in the international literature. The aetiology, clinical findings and therapy of these lesions are discussed. PMID- 8226072 TI - [Treatment of hiatal hernia with Rossetti-Niessen fundoplication by laparoscopy]. PMID- 8226073 TI - [Thoracoscopic resection of a schwannoma]. AB - Schwannomas are most often benign tumors originating from the sheath of peripheral nerves. Most are asymptomatic and detected incidentally on a routine chest X-ray. Resection is generally recommended to confirm the diagnosis. In a 35 year-old patient we resected and removed an intrathoracic schwannoma of the 6th intercostal nerve thoracoscopically. The technique of resection and protected removal is described. PMID- 8226074 TI - [Thoracoscopic treatment of recurrent or persistent spontaneous pneumothorax]. AB - The video shows an endoscopic modification of the established conventional surgical therapy of recurrent or persistent spontaneous pneumothorax: The indications and endoscopic techniques of parietal pleurectomy, ligature of leaking bullae and wedge resection are demonstrated. Our first experience on 50 patients indicates, that minimal postoperative pain and a relatively short hospital stay (mean 3.6 days postop.) are the advantages of minimal invasive techniques also in thoracic surgery. Long-term results are however still lacking. PMID- 8226075 TI - Local and systemic toxicity of intra-hepato-arterial chemotherapy for treatment of unresectable liver metastases of colorectal cancer with 5-Fluorouracil and high dose Leucovorin. AB - We report our preliminary results of an ongoing phase-II-study with special regard to local and systemic toxicity, using a combined regimen of high dose Leucovorin and 5-Fluorouracil applicated intra-hepato-arterially. During 24 chemotherapy cycles we saw only mild to moderate systemic toxicity and there was no chemical hepatitis. First results concerning the response rate are encouraging. The modulation of 5-Fluorouracil (5-FU) with folinic acid represents a significant step in the treatment of colorectal cancer. Administration of chemotherapy through the hepatic artery for liver metastases of colorectal cancer allows high local drug concentration with relative low systemic toxicity. Until now there is little information about the local and systemic toxicity of high dose Leucovorin combined with 5-FU applicated intra-hepato-arterially. We describe our experience with this therapy in a running phase-II-study with special regard to local and systemic toxicity. PMID- 8226076 TI - [Sliding flap-plasty in treatment of anal fistula: a prospective study]. AB - The common therapy for trans- and intersphincteric fistula usually is opening the fistula tract. Healing and rough continence thereafter is uneventful, but when follow-up is intensified and patient are asked for slight soiling and discharge many minor complaints become evident. Therefore we started a series of patients suffering from inter-, trans-, extrasphincteric or supralevatoric fistula with an advancement flap procedure after excision of the fistula, without dissection of any muscle fibre. Up to now 23 patients were operated including 5 with Crohn's disease. 17/23 were healed primarily and 6/23 failed to heal with 5 successful reoperations. 4/5 Crohn's patients operated during quiescent disease are healed. No discharge or soiling was reported after a follow-up time from 5 months to almost 2 years. In conclusion advancement flap gives best results in respect to continence. The flap should include not only mucosa but underlying internal sphincter muscle fibres too. PMID- 8226077 TI - [The disease picture of atypical aortic coarctation]. AB - The atypical coarctation of aorta is a very rare disease. Approximately 1-2% of all coarctations are not localized in isthmus aortae. The etiology of this disease is still unknown. There are existing opinions about acquired and also congenital origin. PMID- 8226078 TI - [Holmium:YAG laser in surgical practice in a multidisciplinary hospital center]. AB - The Holmium:YAG laser has been in use at the regional hospital of Montreux, Switzerland. A balance has been drawn at the end of 9 months. 68 surgical operations have been performed; 23 in abdominal surgery, mainly through laparoscopy, and 45 in orthopaedics using scopic viewing. The Holmium:YAG laser is a modern and up-to-date system. It is well adapted to endoscopic surgery owing to its features, its wide scope and safe use. Its price is rather high if used for laparoscopic cholecystectomy only; but the purchase thereof is proving to be a profitable asset for a multiservice surgical team. PMID- 8226079 TI - [Patient information in surgery]. AB - In a prospective study 50 patients undergoing a hernia repair were asked their opinion about an information sheet which was given to them preoperatively. For 94% this paper is of help; for 82% the written information has to be clarified by an additional personal talk with the doctor. Another possibility, especially good for visualisation, is the video. At the hospital of Davos there are at the moment 7 clips available to explain the treatment and the aftercare after an operation or conservative therapy in traumatology. PMID- 8226080 TI - [Pain syndrome in the right iliac fossa and laparoscopy: routine appendectomy or not?]. AB - Diagnostic laparoscopy is useful in the evaluation of patients with lower right quadrant pain when the diagnosis is uncertain. The object of this study was to determine whether a normal appendix should be routinely removed at laparoscopy. We have decided to do this in children, men under the age of 30 and women of childbearing age. However we do not perform appendectomy in men over the age of 30, women over 50 and in immunodeficient patients. This is a working hypothesis and is the basis of an on-going prospective study in our unit. PMID- 8226081 TI - [Pneumoretroperitoneum: diagnostic and therapeutic approach]. AB - This retrospective study deals with etiology, paths of extension, radiological signs and treatment of pneumoretroperitoneum. The 15 cases presented were seen over a 10-year period. Etiology was infectious in 6 patients, traumatic in 5, and iatrogenic in 4. Treatment applied was surgical in 12 patients, the remaining 3 (who all had their pneumoretroperitoneum secondary to endoscopy) could be handled conservatively. Even though pneumoretroperitoneum is seldom encountered, it should be quickly recognized in order to apply prompt adequate therapy. PMID- 8226082 TI - [Idiopathic spontaneous pneumoperitoneum. Case discussion based on two cases, assessment procedure and therapy and review of the literature]. AB - Idiopathic spontaneous pneumoperitoneum is a rare condition we consider after exclusion of perforation of an intraabdominal viscus and other known causes of free intraperitoneal gas. We will report two cases of idiopathic spontaneous pneumoperitoneum seen during the year 1991 and discuss a rational way to get to the diagnosis. In the absence of peritoneal signs the rather unexpected amount of free intraperitoneal air is a constant finding. Reviewing the literature we found different opinions concerning the treatment of this condition. Regarding the benign character we emphasize a non-surgical management, given the careful exclusion of other, maybe fatal causes of pneumoperitoneum. Diagnostic peritoneal lavage may help to avoid unnecessary laparotomy. PMID- 8226083 TI - [Perforated small intestine diverticulum as a cause of acute abdomen in Ehlers Danlos syndrome]. AB - Based on a rare case of perforated small-bowel diverticulum combined with Ehlers Danlos syndrome (EDS) we investigated in a retrospective study several swiss hospitals for symptomatic small-bowel diverticulosis aiming at any clue on EDS. We came up with 15 more cases, 5 of them with perforation, yet none showed any sign of EDS. A review of the literature on either small-bowel diverticulosis or the intestinal complications of EDS amounted to 131 cases of perforation and 11 cases of symptomatic small-bowel diverticulosis in combination with EDS, 2 of them suffering from perforation. Regarding the rarity of these diseases the number of patients presenting both is surprising. Although the EDS-specific histopathological changes of the skin could not be shown in the specimen of the intestinum, it seems quite reasonable to suggest an increased incidence of diverticulosis in patients with EDS, since the weakness of the connective tissue allows an easy protrusion of the mucosa through the intestinal wall. PMID- 8226084 TI - [Jejuno-jejunal invagination in polypoid lipoma producing renal failure]. AB - This is the case of a 43 years old woman with recurring epigastric pain for 8 years. She was hospitalized emergently for persistent vomiting causing acute renal failure. Numerous radiologic investigations prior to this hospitalisation showed no pathology. The diagnosis was made this time with a small bowel follow through. Intussusception of tumors is a known cause of small bowel obstruction. The clinical picture however, of severe vomiting causing acute renal failure on the basis of a proximal small bowel obstruction is exceptional. PMID- 8226085 TI - [Segmental ischemic colitis in lymphocytic thrombotic venulitis]. AB - This case illustrates the difficulty of diagnosing a colonic stenosis of ischemic origin. A 70-year-old lady presents with abdominal pain, fever and melaena. Lc are 15.2, ESR 39 mm, CEA 2.7 ng/ml. A barium enema shows a stenosis of the transverse colon that is suspicious of neoplasia. At time of operation, an induration of the transverse colon is found with edema of the corresponding mesocolon but no tumour is palpated. A resection of this area is performed and an end to end anastomosis performed. Pathology shows an ischemic colitis secondary to a lymphocytic thrombotic venulitis. The patient is discharged home one month postoperatively. 4 weeks later she is readmitted with the same symptoms. A gastrograffin enema shows a similar stenosis in the transverse colon including the anastomosis. The diagnosis is made of a recurrent ischemic stenosis. The patient improves over a 10-day period of conservative treatment (anticoagulation, TPN, steroids). A control barium enema shows a near resolution of the stenosis. The majority of ischemic colitis are of arterial origin nevertheless ischemic colitis of venous origin exists. The factor causing venous ischemia are not known. It is though thought to be associated with hypersensitivity vasculitis of drug origin. Its initial diagnosis versus neoplasia is difficult but once made there is a good response to a conservative treatment. PMID- 8226086 TI - [Is there a place for thoracic surgery in the peripheral hospital?]. AB - Between 1983 and 1990, 52 patients underwent a thoracotomy in our hospital. After having reviewed these cases, and in particular the 42 who presented a primary lung cancer, we were able to compare our results with those already published. This showed that both our mortality and survival rates were similar to those already published. However, our preoperative evaluation of patients ought to be improved. PMID- 8226087 TI - [Personal experience with preventive use of antibiotics in elective colon surgery. A retrospective study]. AB - We reviewed in a retrospective study all our 616 electively operated colons over 15 years from 1976 to 1990 for morbidity and mortality depending the antibiotic prophylaxis. Interventions were performed on 578 patients with a mean age of 65.5 + 13.7 years; in 38 cases there was an earlier elective colic operation. The sex ratio was 1.26 men to 1 woman. 71.8% of the indications were adenocarcinomas of the colon. All other diagnostics, as well as different types of interventions were included in the study. All interventions was preceded by large bowel preparation. The colic anastomosis were almost always handswen in two layers. Until 1982 systematically, we gave prophylaxis by combination of aminoglycoside with metronidazole or clindamycin, n = 329 (53.4% of 616). We then changed to ceftriaxone, n = 287 (46.6% of 616). Ceftriaxone was applied in single dose (n = 142), in multi doses (n = 71) and combined in almost all cases with metronidazole (n = 74). We found significantly better morbidity and mortality results with ceftriaxone than in the aminoglycoside combined group: The ceftriaxone group had a septicemia or colocutaneous fistulas requiring reoperation incidence of 1.4% (4/287) versus 4.6% (15/329) (p = 0.023). Mortality decreased from 4.9% (16/329) under aminoglycoside combined prophylaxis to 1.74% (5/287) in the ceftriaxone group (p = 0.033). The infection rate of the surgical accesses diminished from 13.1% (43/329) to 8% (23/287) under ceftriaxone (p = 0.043).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226088 TI - [Cholecysto-cholangiography as an alternative to cystic duct cholangiography in laparoscopic cholecystectomy]. AB - Intraoperative cholangiography may be an important adjunct to laparoscopic cholecystectomy in order to prevent bile duct lesions. Laparoscopic cannulation of the cystic duct can be very difficult and time consuming. We therefore developed a simple technique of cholecystocholangiography. The gallbladder is punctured and filled with contrast medium after having localised the cystic duct and put a metal clip as a landmark. The study included 52 patients having either cholecystocholangiography or cystic duct cholangiography. In 13 out of 26 patients (50%) cholecystocholangiography failed because of obstruction of the cystic duct. The cholangiograms showed complete filling of the bile ducts in 5 (19%) and incomplete visualisation in 8 cases (31%). Cystic duct cholangiography showed significantly better results with good delineation of the biliary tree in 19 cases (73%). Five cholangiograms were suboptimal (19%) and only 2 studies failed (8%). The relation between cystic duct and common bile duct was clearly visible in 24 cases (92%) with cystic duct cholangiography compared with 13 cases (50%) with cholecystocholangiography. Cystic duct cholangiography has better success rates in delineating biliary anatomy in order to prevent bile duct injury. Cholecystocholangiography is a good alternative in cases where the cystic duct cannot be initially visualized. PMID- 8226089 TI - [Flexible choledochoscopy: two years experience]. AB - This study is a retrospective review of 258 procedures on the gallbladder and bile ducts (115 without and 143 with flexible choledochoscope). There were 14 bile duct explorations for stone removal without choledochoscopy and 16 with a flexible choledochoscope. Residual stones were found in 2/14 (14.2%) cases without and 1/16 (6.3%) case with choledochoscope. A review of the literature confirms that there are fewer residual bile duct stones following flexible choledochoscopy. There were no complications attributable to the procedure. PMID- 8226090 TI - [Laparoscopic cholecystectomy]. AB - In a community-hospital the problem arises how to improve the knowledge and skill of the surgeons. As mentioned, for some reasons the possibility of training outside the hospital is limited. We get closer to that problem by operating these cases together: two surgeons of two hospitals next each other. An example to describe this problem is the laparotomic cholecystectomy. PMID- 8226091 TI - [Value of laparotomy in massive lower gastrointestinal hemorrhage]. AB - A study of ninety-four patients admitted with massive lower gastro-intestinal bleeding (LGIB) is presented using a systematic diagnostic work-up including angiography, colonoscopy and various investigations, such as scintigraphy, small bowel series and ultrasonography. The bleeding source was identified preoperatively in eighty-five patients. 9 patients had a diagnostic laparotomy and a pathology was found in additional seven. No source was identified in two patients (2.1%) at exploration and "blind" subtotal colectomy was not performed in these two cases as proposed by others. We conclude that a thorough systematic assessment of patients with LGIB is important to localize the bleeding source. Exploratory laparotomy is the final step in few cases (10%) and if no intraoperative source can be identified a "blind" colonic resection should not be performed. PMID- 8226092 TI - Three sickle cell anemia patients each with a different alpha chain variant. Diagnostic complications. AB - We have studied three sickle cell anemia patients who also carried a heterozygosity for one of the following alpha chain abnormalities: Hb G Philadelphia [alpha 68(E17)Asn-->Lys], Hb Montgomery [alpha 48 (CE6)Leu-->Arg], and Hb Chicago [alpha 136(H19)Leu-->Met]. Electrophoretic analyses alone may result in incomplete and incorrect information. Confirmation of the diagnosis of Hb SS or Hb SC disease by one of the fast high performance liquid chromatographic procedures is recommended. PMID- 8226093 TI - Rare beta chain hemoglobin variants found in Swedish patients during HBA1c analysis. AB - Here we report the occurrence of five different beta chain hemoglobin variants not previously described in Sweden. The variants were found during quantification of HbA1c using ion exchange high performance liquid chromatography (HPLC) or isoelectrofocusing. Samples were examined either at protein level by separation of globin chains on C8 reversed phase HPLC, digestion with trypsin or lysylendopeptidase and separation of peptides by C18 reversed phase HPLC, or at DNA level by direct nucleotide sequencing of double-stranded DNA fragments amplified from exon 1 + 2 of the beta-globin gene. The variants were: Hb Raleigh [beta 1 (NA1)Val-->Ac-Ala], Hb J-Baltimore [beta 16(A13)Gly-->Asp], Hb Tacoma [beta 30(B12)Arg-->Ser], Hb K-Ibadan [beta 46(CD5)Gly-->Glu], and Hb Fukuyama [beta 77(EF1)His-->Tyr]. Hb Tacoma, Hb K-Ibadan, and Hb Fukuyama were slightly unstable in the isopropanol test, but no signs of hemolysis were found in the patients who all had normal hematological findings. PMID- 8226094 TI - Hb Shelby [beta 131(H9)Gln-->Lys] in association with Hb S [beta 6(A3)Glu-->Val]: characterization, stability, and effects on Hb S polymerization. AB - When first tested for abnormal hemoglobins, a 2-year-old boy, appeared to have Hb F, Hb S and Hb A2. Confirmatory testing revealed a beta chain variant inherited from his father and beta S from his mother. Analysis of tryptic peptides in conjunction with automated DNA sequence analysis showed that the variant hemoglobin was Hb Shelby [beta 131(H9)Gln-->Lys (CAG-->AAG)]. Heat and mechanical stabilities of various liganded Hb Shelby tetramers were compared to those of Hb A and Hb S. Oxy-Hb Shelby precipitated more readily than oxy-Hb A, but was much more stable than oxy-Hb S during mechanical agitation. In contrast, oxy-Hb Shelby was much less stable than oxy-Hb A and oxy-Hb S following heat treatment. Met-Hb Shelby was most unstable compared to other liganded forms of Hb Shelby, while deoxy- and carbonmonoxy-forms of Hb Shelby showed similar heat-induced precipitation rates. These data indicate that heat instability of Hb Shelby is accompanied by heme oxidation, and that denaturation by mechanical agitation occurs in the absence of heme oxidation. Hb Shelby, like Hb A, can form hybrids with Hb S which participate in polymer formation in vitro. However, Hb S/Hb Shelby hybrids copolymerized with Hb S less than A/S hybrids. Since the patient's MCHC value is normal, this finding coupled with the elevated Hb A2 and Hb F levels, both of which are known to inhibit polymerization of Hb S, may contribute to the patient's mild clinical presentation. PMID- 8226095 TI - Molecular characterization of beta-thalassemia in the United Arab Emirates. AB - This study is to identify the various beta-thalassemic alleles in the United Arab Emirates (UAE), and compare them with the UAE residents from neighboring countries suffering from the same problem. Gene amplification, dot-blot hybridization with synthetic probes, restriction enzyme analyses, and sequencing were the tools used. Thirteen different mutations were observed in the UAE patients and seventeen mutations in the non-locals. The IVS-I-5 (G-->C) Asian Indian mutation was the most frequent mutation in both groups. Homozygous mutations in both groups were relatively higher than double heterozygous mutations. PMID- 8226096 TI - Two new rat alpha-globin sequences as identified by the conserved region PCR. AB - Based on the notion that regions of structural genes which encode critical domains of the corresponding proteins are highly conserved among closely related species, oligonucleotide primers were designed and used to amplify the alpha globin sequence(s) of the Sprague-Dawley (SD) rat. Data of these amplified sequence constructs showed that two new rat alpha-globin specific sequences have been identified. Southern hybridization confirmed the presence of these sequences in the rat genome. PMID- 8226097 TI - Hb Tubingen [alpha 2 beta (2)106(G8)Leu-->Gln] in a Belgian Family. PMID- 8226098 TI - Hb J-Guantanamo [alpha 2 beta 2 128(H6)Ala-->Asp] found in a Japanese family. PMID- 8226099 TI - A novel deletion in a Turkish beta-thalassemia patient detected by DGGE and direct sequencing: FSC 22-24 (-7 bp). PMID- 8226100 TI - Lysosomal storage diseases: mechanisms of enzyme replacement therapy. AB - Lysosomal diseases result from deficiency of one of the many enzymes involved in the normal, step-wise breakdown of macromolecules. Studies in vitro have shown that cells from enzyme-deficient patients can be corrected by an exogenous supply of the missing enzyme. This occurs by receptor-mediated endocytosis of normal enzyme added to tissue culture medium and also by direct transfer from normal leukocytes during cell-to-cell contact. Immunohistochemical analysis has revealed that these processes have similar pathways of intracellular transport of the acquired enzymes, which ultimately reach mature lysosomes in the recipient cells. Moreover, recent studies suggest that both mechanisms are important in the therapy of lysosomal storage diseases by bone marrow transplantation. Advances in gene technology are likely to improve the successful treatment of these disorders, by facilitating the large scale production of clinically effective proteins and also by enabling the stable and safe introduction of normal lysosomal genes into cells of affected patients. PMID- 8226101 TI - Immunohistochemical localization of transforming growth factor alpha in the major salivary glands of male and female rats. AB - The three major salivary glands of normal male and female Fischer 344 rats of different ages were examined for the localization of epidermal growth factor (EGF) and transforming growth factor alpha (TGF alpha) by immunohistochemical staining. EGF was demonstrated only in the granulated convoluted tubule (GCT) cells of the submandibular gland, the results confirming the previous reports, and most abundantly in adult males and pregnant females. TGF alpha stain was localized in all three glands and was found throughout the entire duct system, excluding acinar cells. The myoepithelial cells of the sublingual gland were also reactive with the TGF alpha antibody. The specificity of the staining was confirmed by negative staining reaction with the absorbed antibody and by radio immunoassay and Western blot methods. This is the first report describing the presence of TGF alpha in the rat salivary glands. PMID- 8226102 TI - Mast cell immunohistochemistry: non-immunological immunostaining mediated by non specific F(ab')2-mast cell secretory granule interaction. AB - During investigations of murine and human mast cell immunoreactivity with potential anti-interleukin-4 antibodies, non-specific, non-immunological labelling of mouse and human mast cells became apparent. Non-specific, non immunological labelling was identified by (i) immunolabelling of mast cells when using control isotype primary antibodies, (ii) ability of conjugated secondary antibodies to label mast cells without prior mast cell exposure to a primary antibody, (iii) extinction of the non-specific labelling and retention of specific labelling when the pH of the diluting and washing buffers is shifted from pH 7.2 to pH 6.0, and (iv) reduction/extinction of the labelling when the antibodies are pre-incubated with soluble heparin prior to immunostaining. The site of the reactivity on the electron microscope level was shown to be confined to the mast cell secretory granules. The results of this study support the hypothesis that non-specific labelling of mast cells results from an ionic interaction between the F(ab')2 segments of antibodies and the heparin constituent of the mast cell secretory granules. This study points out the necessity of stringent controls when using immunohistochemistry to determine mast cell reactivity to various antibodies. PMID- 8226103 TI - Electron microscopic immunocytochemical demonstration of blood-retinal barrier breakdown in human diabetics and its association with aldose reductase in retinal vascular endothelium and retinal pigment epithelium. AB - Light-microscopic immunohistochemical staining for albumin has been used to localize sites of blood-retinal barrier (BRB) breakdown in ocular disorders, but the mechanism for BRB compromise cannot be resolved at this level. Using eyes up to 2 days post-mortem from normal patients or from patients with diabetic retinopathy, or other disorders known to cause BRB failure, electron-microscopic immunocytochemistry reveals focal breakdown of the inner BRB, comprised of the retinal vascular endothelium (RVE), which appears to be mediated by diffuse permeation of the RVE cells and by vesicular transport. Permeation of the retinal pigmented epithelial (RPE) cells that comprise the outer BRB also occurs, but there is no evidence of opening of tight junctions between RVE or RPE in any of the disorders evaluated. Increased aldose reductase (AR) expression in the RVE and RPE cells of diabetics as well as in the perivascular retinal astrocytes, which interact with RVE cells to establish the inner BRB, suggests that AR activity and the subsequent intracellular accumulation of sorbitol in these cell types may impair the function of the BRB in diabetes. PMID- 8226104 TI - Alpha-spectrin in detergent-extracted whole-mount cytoskeletons of chicken embryo heart fibroblasts. AB - The distribution of alpha-spectrin, and its relation to other cytoskeletal structures and to the plasma membrane, was studied in detergent-extracted whole mount cytoskeletons of chicken embryo heart fibroblasts by using immunogold labelling and electron microscopy (IEM). The cell surface was labelled with gold conjugated wheat germ agglutinin (WGA-gold), microtubules with anti-tubulin antibodies, and spectrin by using antibodies raised to chicken erythrocyte alpha spectrin. Additionally, the effect of fixation and drying on the labelling pattern was evaluated. In electron microscopy, a three-dimensional filamentous network was observed in detergent-extracted whole-mount preparations. Filaments of diameter 7-10 nm and 15 nm, microtubules of diameter 30 nm, and filament bundles (40-50 nm in diameter) were seen. In IEM, alpha-spectrin was seen on the surface of the cytoskeletal network, especially along the thick filament bundles. In some cells, a distinct membrane skeleton which was labelled with alpha spectrin antibodies, was seen in close association with the cytoskeletal network. The cells which were labelled first with WGA-gold, and then permeabilized, fixed and labelled with alpha-spectrin, showed a co-localization of the WGA binding sites and alpha-spectrin along the surface of the filament bundles. Reversing the order of the staining, such that fixation was done before WGA labelling and permeabilization, led to a greatly diminished labelling for alpha-spectrin and less pronounced co-localization of spectrin and WGA. Comparison of the conventional critical point drying method with Peldri II, a novel drying agent, indicated a better stability of the cellular structures under the electron beam when Peldri II was used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226105 TI - A letter to young researchers in the biomedical sciences. PMID- 8226106 TI - Stability of elastin in the developing mouse aorta: a quantitative radioautographic study. AB - Elastic lamina growth during development and the ultimate stability of elastin in the mouse aortic media was investigated by light and electron microscopic radioautography. Following a single subcutaneous injection of L-[3,4-3H]valine at 3 days of age, animals were killed at 9 subsequent time intervals up to 4 months of age. One day after injection, radioautographic silver grains were primarily observed over the elastic laminae; however, silver grains were also seen over the smooth muscle cells and extracellular matrix. By 21 to 28 days of age, the silver grains were almost exclusively located over the elastic laminae. From 28 days to 4 months of age, the distribution of silver grains appeared relatively unchanged. Quantitation of silver grain number/micron2 of elastin showed a steady decrease in the concentration of silver grains associated with the elastic laminae from 4 to 21 days of age. After this time, no significant difference in silver grain concentration was observed. Since the initial decrease in grains/micron2 of elastin corresponds to a period of rapid post-natal growth, the decrease is likely to be a result of dilution of the radiolabel due to new elastin synthesis. With the assumption that little or no significant turnover occurs during this time, a constant growth rate of 4.3% per day was predicted by linear regression analysis. Since no significant difference in the concentration of silver grains was observed from 28 to 118 days of age, no new growth or turnover of elastin can be said to occur during this time period. This is supported by the observation that animals injected with radiolabeled valine at 28 days and 8 months of age showed no significant incorporation of radiolabel into the elastic laminae. The results from this study present the first long-term radioautographic evidence of the stability of aortic elastin and emphasize that initial deposition of elastin and proper assembly of elastic laminae is a critical event in vessel development. PMID- 8226107 TI - Distribution of actin in cone photoreceptor synapses. AB - The invaginated photoreceptor terminal harbours a great number of postsynaptic dendrites that contact the photoreceptor terminal at morphologically distinct synapses. In this paper we studied the subcellular distribution of actin in cone photoreceptor synapses of light-adapted goldfish and guppy retinas and its relation to ribbon and spinule synapses. In the outer plexiform layer, fluorescent staining for actin by antibodies and also by fluorescently labelled phalloidin was discontinuous and showed large dot-like immunoreactive structures (3-5 microns in diameter). Immunogold electron microscopy of light-adapted retinas isolated 3 h after the onset of light demonstrated that these dot-like structures corresponded to dendritic profiles of bipolar and horizontal cells within the synaptic cavity of cones. A much less intense staining was observed within the presynaptic cone photoreceptor terminal itself. Immunogold particles were also found overlying the tips of horizontal and bipolar dendrites that form the triadic postsynaptic complex of the ribbon synapse. In contrast to the triads light-induced terminal outgrowings of horizontal cells, also known as spinules, remained largely unlabelled. These observations suggest that actin plays a role in maintenance and/or certain steps of the formation of the postsynaptic dendritic complex. PMID- 8226108 TI - Experimentally induced colon cancer metastases in rat liver increase the proliferation rate and capacity for purine catabolism in liver cells. AB - Metastases in rat liver were generated experimentally by intraportal injection of colon cancer cells to investigate the effects of cancerous growth on the metabolism of surrounding liver tissue. Maximum activities (capacity) of glucose 6-phosphate dehydrogenase, phosphogluconate dehydrogenase, lactate dehydrogenase, succinate dehydrogenase, alkaline phosphatase, 5'-nucleotidase, xanthine oxidoreductase, purine nucleoside phosphorylase and adenosine triphosphatase have been determined. Two types of metastases were found, a small type surrounded by stroma and a larger type in direct contact with hepatocytes. Both types affected the adjacent tissue in a similar way suggesting that the interactions were not mediated by stroma. High capacity of the degradation pathway of extracellular purines released from dead cells of either tumours or host tissue was found in stroma and sinusoidal cells. Metastases induced both an increase in the number of Kupffer cells and proliferation of hepatocytes. The distribution pattern in the liver lobulus of most enzymes investigated did not change distinctly. However, activity of alkaline phosphatase, succinate dehydrogenase and phosphogluconate dehydrogenase was increased in hepatocytes directly surrounding metastases. These data imply that the overall metabolic zonation in liver lobuli is not dramatically disturbed by the presence of cancer cells despite the fact that various metabolic processes in liver cells are affected. PMID- 8226109 TI - Connexin43 in rat pituitary: localization at pituicyte and stellate cell gap junctions and within gonadotrophs. AB - Immunohistochemical methods were employed to investigate the cellular and ultrastructural localization of the gap junction protein connexin43 (Cx43) in rat pituitary. Western blots of pituitary homogenates probed with anti-Cx43 antibodies showed the presence of Cx43 in both anterior and posterior pituitary lobes. By light microscopy (LM), Cx43-immunoreactive (Cx43-IR) puncta were found in all areas of the posterior lobe, but at greater concentrations in peripheral regions of this structure. By electron microscopy (EM), immunogold labelling for Cx43 was seen at gap junctions between thin cytoplasmic processes of pituicytes. No immunoreactivity was detected in the intermediate lobe. The anterior lobe contained puncta similar to but more sparsely scattered than those in the posterior lobe, and by EM analysis these were demonstrated to correspond to labelled gap junctions between stellate cells. In addition, anti-Cx43 antibodies produced intracellular labelling in a small percentage of endocrine cells, which were distributed throughout the anterior lobe and determined by double immunostaining methods to be cells containing luteinizing hormone. By EM, labelling within these cells was associated with predominantly large secretory granules and other loosely organized organelles. The results indicate that gap junctions in the pituitary are composed of Cx43 and that this or a related protein may have a novel intracellular function within gonadotrophs. PMID- 8226110 TI - Immunolocalization of calcium vector protein and its target protein in amphioxus. AB - Three proteins, sarcoplasmic Ca(2+)-binding protein (SCP), Ca2+ vector protein (CaVP) and its target protein (CaVPT), are found abundantly in the higher invertebrate amphioxus. Whereas the function of SCP is likely to be related to Ca2+ and Mg2+ buffering, that of the latter two proteins, apparently linked together, is still not clear. In this study, affinity-purified polyclonal antibodies to these three proteins were used to study the extractability under physiological ionic conditions, the distribution in different tissues and the immunocytochemical localization in striated muscle. Our data show that SCP is essentially cytosolic whereas CaVP and CaVPT are partially associated with non soluble components in amphioxus tissues. The tissue distribution, studied in transverse sections, shows that SCP is merely confined to striated muscle, whereas CaVP and CaVPT are also abundant in other tissues such as the spinal chord and the gonads. Thus the protein pair CaVP/CaVPT is likely to serve a general role in many tissues; however, no strict correlation was found in the distribution of the latter two proteins, suggesting that they may function independently. The detailed cytochemical localization of the three proteins in longitudinal sections of striated muscle revealed a discrete striation pattern in addition to a diffuse background. For SCP these striations are coincident with the Z line. The immunostaining for CaVP shows intense striations at the level of the Z lines alternating with weak striations at the M lines. For CaVPT the striations at the Z and M line are more or less of equal intensity, leading to a pattern with a 1 micron periodicity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226111 TI - Demonstration of feline and canine platelet glycoproteins by immuno- and lectin histochemistry. AB - Canine and feline platelet cytocentrifuge preparations (CCPs), cryostat and paraffin-embedded bone marrow sections were used in this study. We evaluated whether platelets, megakaryocytes and megakaryocyte precursor cells could be labelled by monoclonal antibodies (Y2/51, CLB-thromb/1, HPL1) against human platelet membrane glycoprotein GP IIIa and the GP IIb/IIIa complex or by the following 10 biotinylated lectins: concanavalin A (Con A), Lens culinaris agglutinin (LCA), Pisum sativum agglutinin (PsA), wheat germ agglutinin (WGA), peanut agglutinin (PNA), Phaseolus vulgaris lectin (PHA-L), Ricinus communis agglutinin 120 (RCA120), Ulex europaeus agglutinin-I(UEA-1), soybean agglutinin (SBA) and Dolichos biflorus agglutinin (DBA). Monoclonal antibodies Y2/51 and HPL1 cross reacted with platelets and megakaryocytic cells from both species, whereas CLB-thromb/1 was unreactive with canine preparations. Only Y2/51 labelled megakaryocytic cells in paraffin-embedded samples. LCA, PSA, WGA and PHA-L labelled feline and canine platelets and different numbers of morphologically identifiable megakaryocytes and numerous other, mostly myeloid, cells. Immunoblots of dog and cat platelet lysates using Y2/51 visualized a single protein of 95 kDa (unreduced), a mol.wt value within the range of those reported for GP IIIa. Some of the platelet (but not necessarily megakaryocyte) glycoproteins reacting with LCA, PSA and WGA could be identified in lectin blots following one- or two (nonreduced/reduced)-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Thus in dogs and cats, the immunohistochemical detection of GP IIIa (and eventually GP IIb/IIIa) rather than lectin binding patterns could be important for the diagnosis of megakaryoblastic leukaemias. PMID- 8226112 TI - Determination method for methyl iodide using diffusive sampler and capillary GC with thermal desorption injector and FID. AB - A determination method for methyl iodide in the work place has been developed, using a diffusive sampler and a capillary gas chromatograph with a thermal desorption cold trap injector and an FID. The sampler consists of a glass tube (6 mm o.d., 4 mm i.d., 16 cm length) packed with porous polymer beads. The diffusion path of the sampler is made of a 3 mm i.d., 52 mm length poly (tetrafluoroethylene) tube inserted in the glass tube. The sampling rate of the sampler for methyl iodide was 4.6 ml/h, and variations caused by temperature, relative humidity, and wind speed were found to be small and negligible. The sample could be stored for at least two weeks without significant loss. A time weighted average concentration of methyl iodide in the range of sub ppm to ppm for two to six hours can be measured by this method. The average concentration of methyl iodide for a period of a few seconds can also be measured by grab sampling, i.e., injecting 20 ml of air directly into the sampler. PMID- 8226113 TI - Interrelation between urinary delta-aminolevulinic acid (ALA), serum ALA, and blood lead in workers exposed to lead. AB - Using a fluorometric HPLC method, we determined delta-aminolevulinic acid (ALA) in sera and urine samples from 16 lead workers with blood lead levels ranging from 19 to 107 microG/100 ml. The concentration of ALA in serum correlated highly with the urinary ALA concentration (gamma = 0.957 for ALA mg/1; gamma = 0.967 for ALA mg/g creatinine). The ALA concentrations in the serum of lead workers ranged from 11 to 151 micrograms/1 with a mean of 51 micrograms/1. In addition, the concentrations of urinary ALA (mg/g creatinine) and serum ALA micrograms/1) had a strong correlation with blood lead concentrations (gamma = 0.838 and 0.892, respectively). These data indicate that the measurement of serum ALA, as well as urinary ALA, is very useful for the biological monitoring of occupational lead exposure. PMID- 8226114 TI - Testicular toxicity of thiram in rat: morphological and biochemical evaluations. AB - Tetramethylthiuram disulphide (Thiram), a dithiocarbamate fungicide was administered to male rats through gavage at dose 5, 10 and 25 mg/kg/day for 30, 60 and 90 days. A significant increase in relative testes weight (GSI) was observed at dose (25 mg/kg/d) after 90 days exposure, associated with mild pathomorphological changes. This included degeneration of seminiferous tubules with sloughing of germinal cells and accumulation of edematous fluid in tubular lumen. The activity of testicular enzymes such as lactate dehydrogenase (LDH), alkaline phosphatase (ALP), glucose 6-phosphate dehydrogenase (G-6-PDH) were increased significantly in dose dependent manner. The activities of succinate dehydrogenase (SDH) and acid phosphatase (ACP) decreased significantly while serum cholesterol and testicular free sialic acid were increased at all dose levels. The alterations in the activity of testicular enzymes associated with marked increase in GSI and pathomorphological changes suggest that thiram induces testicular dysfunctions in rats. PMID- 8226115 TI - Development of a system for analyzing working postures. AB - In order to assist in the analysis of working postures which might cause lumbago in the field of industrial health, a system was developed for the synchronous recording and analyzing of postures, work content and physiological data. The system is composed of a portable unit for recording 3 channels of goniometers, 1 channel of inclinometer and 1 channel of surface electromyogram, a video camera for recording work content, a host computer and some peripheral devices for analyzing the data from the portable unit and video camera. In this system, postures are automatically classified from data on joint angles and upper body inclination angle by using a discriminant function. The joint angles are measured by the goniometers using rubber optical fibers. The angle of upper body inclination is obtained by the inclinometer using a magnetic resistance sensor. In addition to the work content and video images, the postures and the electromyogram can be analyzed and confirmed by simultaneous display of the data on the host computer screen. Based on the trial use of this system in a work model of manual baggage handling, it was proved useful for evaluating in detail the workload caused by working postures. PMID- 8226116 TI - [Pathogenesis of sinusitis. Not exclusively a ventilation problem]. PMID- 8226117 TI - [The Klockhoff test in Meniere disease]. PMID- 8226118 TI - [Noise-induced hearing loss--also in secretaries and due to noise of leisure activities]. PMID- 8226119 TI - [Treatment of perennial non-allergic rhinopathy with capsaicin]. AB - Twenty-seven patients suffering from perennial non-allergic rhinitis were treated with capsaicin. The drug was applied intranasally seven times during intervals ranging from 4-7 days. The regimen involved application of a 0.5 ml of a 10 micromolar capsaicin solution on the first day of treatment, 0.5 ml of a 30 micromolar solution on the 2nd and 3rd days of treatment and 0.5 ml of a 100 micromolar solution on the 4th to 7th days of treatment. Two patients dropped out, one because he developed an exanthema of both forearms and the other for unknown reasons. Other side-effects observed were epistaxis (1 case) and increased dryness of the nasal mucosa. Twenty-two patients had experienced nasal obstruction, hypersecretion, nasal itching, sneezing, mucosal dryness and headache before and 6 months following capsaicin application and judged the success of therapy 6 months following treatment. Three patients did not take part in the follow-up. Fifteen of 27 patients felt no change had occurred in nasal complaints, while 7/27 scored significant improvement. Additionally, 11/27 subjects would not undergo this treatment a second time because it had been unsuccessful and/or painful. The mean symptom scores for nasal obstruction, hypersecretion, itching, dryness and headache revealed no significant changes before and 6 months following treatment with capsaicin. Blockers did not benefit from capsaicin treatment. The best change to benefit from intranasal capsaicin application involved patients with nasal hypersecretion and sneezing without substantial obstruction. PMID- 8226120 TI - [The pathogenesis of sinusitis in intensive care patients]. AB - Acute sinusitis is a frequent complication in ventilated intensive care patients and may be a possible source of pneumonia or septicemia. A study of 49 ventilated intensive care neurosurgical patients without previously known disease of the paranasal sinuses or midface fractures was conducted retrospectively from 1989 1990. The kind of intubation used (naso- or orotracheal) was taken into account and the period of ventilation examined in order to determine the genesis of inflammatory changes in the paranasal sinuses (as defined by computed tomography). Intensive care patients suffering from sinusitis showed a characteristic early opacity of the sphenoid sinuses, with lesser involvements in the ethmoid and maxillary sinuses. Only in rare cases and after very long periods of ventilation were the frontal sinuses found to be opaque. Nasotracheal ventilation was observed to produce an earlier attack on the intubated ipsilateral sinuses. These findings indicate that nasotracheal intubation should be avoided if possible or the method of intubation changed as early as feasible. If conservative measures fail sinusitis should best be treated by means of endonasal microsurgical open sinostomy. PMID- 8226121 TI - [Revascularized over-long jejunum segments in single stage reconstruction of voice and deglutition function after total laryngopharyngectomy]. AB - After surgery of advanced larynx and pharynx malignancies, a main problem is reconstruction of swallowing and voice. With microvascular small bowl transplantation, we have been able to use a 30 cm segment of jejunum in a one stage procedure to reconstruct extended defects of the oropharynx, and total by hypopharynx, as well as to create a speaking-syphon as devised by Ehrenberger. The surgical technique is described and we now report the results achieved in our first five patients. This new method enables pharyngectomized and laryngectomized patients to speak and to eat without any aspiration after about two weeks. This rapid and functionally encouraging rehabilitation means a significant improvement in a patient's quality of life and is further justification for the great effort required in using this method. PMID- 8226122 TI - [Jaffe-Lichtenstein fibrous dysplasia as a cause of progressive protrusio bulbi and temporary vision loss]. AB - We report the rare clinical and morphological features found in a patient with fibrous dysplasia of the paranasal sinuses. In general, diagnosis of tumor sinuses can be made without difficulty and can be made on the basis of three criteria: clinical examination, radiographic findings and histologic examination of a biopsy. Nevertheless, if rare morphological findings occur, obtaining the correct diagnosis can be complicated. Therefore, we report our experiences in managing a 22 year old woman with fibrous dysplasia located in the paranasal sinuses that resulted in proptosis and temporary blindness. The diagnostic and therapeutic implications are elucidated. PMID- 8226123 TI - [Early cancer detection in the area of the upper aerodigestive tract. II]. PMID- 8226124 TI - [Masking. What is it?--When can it be used?]. PMID- 8226125 TI - [Hearing loss caused by nocturnal hypotension?]. PMID- 8226126 TI - [Reconstruction of the sound transmission system. Which implant materials are best according to the current state of knowledge?]. PMID- 8226127 TI - [Comparison of the prognostic value between the nerve excitability test and maximum stimulation test in evaluation of Bell's palsy]. AB - In the present comparative study we applied the nerve excitability test (NET) and the maximum stimulation test (MST) to 131 patients with idiopathic facial nerve palsy. Our aim was to determine their value in the evaluation of facial nerve function. Results showed that the percentage of error of the NET in the prognosis of facial nerve palsy was 16%. In contrast, the MST was simple in its application and had a high percentage of reliability (97%). However, it remains a qualitative method, since it is dependent on the examiner's observations. PMID- 8226128 TI - [Preventive antibiotic administration in rhinoplasty. Tissue level and pharmacokinetics with reference to rhinoplasty techniques]. AB - Prophylactic antibiotics (cefuroxime 1500 mg) were administered during induction of anesthesia to 34 patients undergoing nasal surgery. The infusion ended at the time of mucosa incision (mean infusion time: 22 min). A hemitransfixion incision was followed by elevation of the mucoperichondrium on both sides of the septum. The blood supply of the cartilage was thus diminished essentially. In the septal cartilage, the concentration of cefuroxime 68 min after the administrations of antibiotic was 28.1 micrograms/g; this concentration was 19.8 micrograms/g in the septal bone at 74 min. The mean peak plasma concentration was 158 micrograms/ml and the end half-life elimination time 77 min. High antibiotic concentrations in the cartilage resulted from passive diffusion through the septal mucosa and occurred at a mean time of 22 min. The time between the start of infusion at the induction of anesthesia and interruption of the septal blood supply by separating of the septum from overlying soft tissues was sufficient for developing high antibiotic concentrations. During nasal surgery the routine approach to the septum had no influence on the efficacy of antimicrobial prophylaxis. PMID- 8226129 TI - [Animal experiment studies using a low-frequency, phase-delayed masker in inner ear diagnosis]. AB - Endolymphatic hydrops is usually diagnosed on the basis of clinical symptoms although objective parameters for this disease can be difficult to differentiate from other disorders of the inner ear. Low-frequency acoustic biasing using a highly intensive phase-shifted low-frequency masker was studied according to its ability to determine disorders of cochlear micromechanics in diseases caused in animal models. Ossicular disruption, experimentally induced early endolymphatic hydrops and gentamycin-induced inner ear damage could be objectively determined histologically and electrophysiologically. Statistical analysis proved that the low-frequency acoustic biasing technique allowed electrophysiologic differentiation of the groups studied that was independent from known clinical parameters. To substantiate the group differences by variance analysis, an intensity of at least 100 dB SPL was required for the 52 Hz masker, and the difference in relation to the test-tone intensity had to be 10 or 20 dB SPL. The early stages of experimentally-induced endolymphatic hydrops could be identified by means of a low-distinct modulation span of the compound action potential amplitude (< 50 uV) and a reduced positive summating potential amplitude with a phase-shifted maximum value compared to the control groups. During the test phases the cochlear microphonic potentials showed only slight suppression of their amplitudes and no modulation effect when compared to the control groups. The modulation behavior of the responses could be correlated with the histologic and supposed micromechanic states of the basilar membrane and organ of Corti, thus allowing validation of the measuring technique. PMID- 8226130 TI - [Acute hearing loss caused by arterial hypotension]. AB - The aim of this study was to confirm the hypothesis based on clinical observations of a causal relationship between arterial hypotension and sudden hearing loss of lower frequencies. A noninvasive system was used that enables repeated blood pressure measurements at programmable time intervals. With this method 24-hour blood pressure monitoring was performed on 81 patients with sudden inner ear hearing loss proved by audiological tests (and in part defined by magnetic resonance imaging). The profiles of the circadian blood pressures recorded actually revealed a significant incidence of low-frequency hearing loss of up to 30 dB HL in the patient group suffering from arterial hypotension compared to those showing normal blood pressure or hypertension. In the patient group designated arterial hypotension may be the probable cause for sudden hearing impairments and should be regarded as a circulatory disorder with certain implications for management. PMID- 8226131 TI - [Teeth as prostheses materials for reconstruction of disrupted ossicular chain]. AB - Since 1988 we have routinely used prostheses made from teeth for ossicular reconstruction and have now done more than 400 implants. Short-time results with at least 1 year implantation were studied in a retrospective analysis of 319 cases with a review of 88 patients. Eighty percent of the implants were covered by an intact tympanic membrane, while in 10% the prosthesis was completely uncovered. This apparent perforation was not due to a material-related extrusion, but was caused by persistent ventilation dysfunction of the middle ear. This was associated with a progressive retraction of the tympanic membrane, which could be seen in all of these cases. The overall acoustic results were disappointing and hearing gain was only 8.7 dB. However, these results were due to the unsolved problems of chronic middle ear inflammation and were not influenced by the dental material. We have been very satisfied with the teeth as implant material, as they can be autoclaved (eliminating problems due to infection) without any loss of mechanical strength. Furthermore, their practically cost-free production has a definite financial advantage over the expensive, commercially-available implants. The prostheses can be sculptured by the young resident during his otologic training, with this experience then resulting in better and faster tympanoplasties. PMID- 8226132 TI - [Infected cystadenolymphoma. A possible reason for clinical and cytologic diagnostic error "carcinoma"]. AB - Inflamed cystadenolymphomas may mimic malignancies both clinically and cytologically. Inflammation-induced facial nerve paralysis, as a clinical sign, and atypical epithelial cells (arising from the inflamed cystic portion of the tumour), as a cytological marker of malignancy, are responsible for this false diagnosis. The diagnostic problems encountered are demonstrated by two case reports. PMID- 8226133 TI - [Treatment of zoster oticus with acyclovir. Report from general practice and a small hospital department]. AB - Findings in patients having zoster oticus infection are reported following treatment in an oto-rhino-laryngological practice connected to a small hospital department. Treatment involved intravenous administration of acyclovir and-in most cases-additional infusions of dextran, cortisone and naftidrofuryl, as adapted from a drug regimen proposed by Stennert. Although treatment was not effective in all cases, several very good results and some partial results were found that justified continued use of this combination therapy. PMID- 8226134 TI - [Tympanic effusion]. AB - Since epidemiologic studies have shown, that the majority of middle ear effusions disappear spontaneously and sequelae a less frequent than expected, a more conservative treatment strategy is advisable. In this review pathogenetic factors are discussed and a treatment policy is recommended as it appears logic to the authors. It is stressed that exact audiologic data are difficult to obtain in children. If an effusion persists at least for 3 months, adenoidectomy, paracentesis and aspiration of the middle ear fluid has to be considered. The insertion of middle ear ventilation tubes is not the first step in our treatment. Special strategies for treatment have to be applied in some groups such as children with Francheschetti or Down Syndrome and children with cleft palates as well as in children with retardation in speech development. PMID- 8226135 TI - [Welding drop injury]. PMID- 8226136 TI - Analysis of accessory signaling in human T-cell clones. AB - Human T cells, when activated, express small but detectable levels of MHC class II on their surface and as a result have the potential to present antigens in the context of MHC class II molecules. There are reports demonstrating MHC class II restricted antigen presentation by human T-cell clones. In this report, we show evidence for one such clone that can present peptide antigen to itself. This clone, HA1.7 ml, is clearly different in its accessory signaling requirements from the parental clone HA1.7, and shows a decrease in the surface level of CD54. We have analyzed the accessory signaling abilities of T cells by using HA1.7 ml as a responder population, and demonstrate that the accessory signaling potential of T-cell clones is sufficient to bring about activation of HA1.7 ml but not of HA1.7. This accessory signaling ability is, however, different from that of B cells in providing bystander accessory signals. T-cell APCs cannot provide bystander accessory signals for the proliferative response of HA1.7 ml, nor can they block the induction of tolerance in this subline, whereas bystander B cells can mediate both events to a limited extent. Thus, there are significant differences in the accessory signaling abilities of T cells as APCs as compared with classic APCs such as B cells, independent of their ability to generate peptide-MHC complexes. It also appears that although HA1.7 ml can be activated by self-presentation of antigen, it is still susceptible to the induction of tolerance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226137 TI - Host T-cell primary allosensitization to MHC class-I- and class-II-expressing human cardiac myocytes requires the presence of a second signal. AB - Normal FHCMs, or transformed cell lines derived from FHCMs, such as W1, even after induction of MHC antigens by pretreatment with IFN-gamma, failed to induce proliferation of allogeneic human PBMCs in vitro. To test the hypothesis that antigen-specific T-cell activation and proliferation require not only the binding of the TCR with its ligand, the MHC molecule, but also a second signal that involves the interaction of T-cell surface molecules with their natural ligands on the stimulating cells, a mAb against CD28 was used. Cocultures of allogeneic PBMCs with IFN-gamma-pretreated irradiated FHCMs or the W1 cell line in microtiter plates containing immobilized anti-CD28 mAb induced marked stimulator cells MHC class-II-specific proliferative responses. The W1 cell line and FHCMs failed to express detectable levels of the BB1/B7 molecule (the natural ligand for CD28) as determined by flow microfluorometry or mRNA levels coding for BB1/B7 as determined by RT-PCR. These data suggest that one of the probably reasons for the failure of MHC-expressing cardiac myocytes to induce allogeneic activation is the absence of costimulatory signals. PMID- 8226138 TI - Allelic typing of the HLA-DR4 group by polymerase chain reaction--single-strand conformation polymorphism analysis. AB - The technique of SSCP was applied to the molecular subtyping of the HLA-DR4 group of alleles. SSCPs were defined in 16 cell line DNAs, representing the alleles DRB1*0401-*0411 (except DRB1*0409), using group-specific PCR amplification, nondenaturing polyacrylamide gel electrophoresis, and silver staining. Three sets of gel electrophoresis conditions were necessary to differentiate the 10 alleles tested. The SSCPs defined in the cell line DNAs were validated by analysis of 145 unrelated HLA-DR4-positive individuals previously genotyped for DRB1*04 alleles by sequence-specific oligotyping. This study demonstrates the suitability of the SSCP technique for defining DRB1*04 alleles, particularly when a small number of samples require typing. PMID- 8226139 TI - Combinations of specific DRB1, DQA1, DQB1 haplotypes are associated with insulin dependent diabetes mellitus in Sardinia. AB - The Sardinian population has an extremely high incidence of IDDM (30.2 of 100.000 in the age group of 0-14 years). This study reports the molecular characterization of HLA class II genes in 120 IDDM sporadic patients and 89 healthy subjects of Sardinian origin. Compared with other Caucasians, both Sardinian patients and controls had an unusual distribution of haplotypes and genotypes. In particular, there was a high gene frequency of the DRB1*0301, DQA1*0501, DQB1*0201 susceptibility haplotype both in patients (0.58) and controls (0.23) while a reduction of the DRB1*1501, DQA1*0102, DQB1*0602 protective haplotype (0.03) was observed in the healthy population. This distribution may partially explain the high incidence of IDDM reported in Sardinia. The analysis of the DQ beta 57 and DQ alpha 52 residues showed that the absence of Asp 57 and the presence of Arg 52 were associated with IDDM in a dose response manner. On the other hand, we found that (a) a very similar distribution of these residues was found when comparing Sardinians with another healthy Caucasian population from the same latitude but with a lower rate of IDDM incidence; (b) several genotypes encoding the identical DQ alpha 52/DQ beta 57 phenotype carried very different relative risks; and (c) the DRB1*0403, DQA1*0301, DQB1*0304 haplotype (DQ beta 57 Asp-neg and DQ alpha 52 Arg-pos) was found in 40% of the DR4-positive controls but not in patients (p = 0.00034), while the DRB1*0405, DQA1*0301, and DQB1*0302 haplotype carrying the same residues at the same positions was found in 70% of the DR4-positive patients and in only one control (p = 0.00003). These findings suggest that IDDM susceptibility cannot be completely explained by the model in which only DQ alpha 52 and DQ beta 57 residues are taken into account. PMID- 8226140 TI - Interruptions of high-dose radiation therapy decrease long-term survival of favorable patients with unresectable non-small cell carcinoma of the lung: analysis of 1244 cases from 3 Radiation Therapy Oncology Group (RTOG) trials. AB - PURPOSE: To determine if prolonged treatment time adversely affects survival for patients with inoperable non-small cell carcinoma of the lung. METHODS AND MATERIALS: Patients enrolled on three randomized studies (RTOG 8311, 8321, 8403) between 1983-1989 formed the database. Previous analyses found that the addition of thymosin (8321) or prophylactic cranial irradiation (8403) failed to prolong survival: both studies used thoracic irradiation with standard fractionation to 55-60 Gy in 30 fractions. In 8311, patients were treated by hyperfractionated radiation therapy to randomly assigned total doses of 60.0 Gy, 64.8 Gy, 69.6 Gy, 74.4 Gy or 79.2 Gy, 1.2 Gy twice daily, 5 days per week. Patients analyzed received +/- 4% of the assigned total dose and lived > 90 days (to ensure that all patients would have completed treatment). Completion < 5 days beyond protocol specifications was classified as "per protocol." Elapsed treatment time exceeding specifications by 5-9 days was a minor deviation, 10-13 days was a major deviation-acceptable, and > or = 14 days was a major deviation-unacceptable. Absolute survival was the endpoint to evaluate the effect of delays. The log rank statistic was used to test for survival differences in the univariate setting, the Cox regression model was used in the multivariate setting. RESULTS: Of 293 patients treated with standard fractionation, eight (2.7%) had deviations from the specified treatment time (six minor, two major-acceptable). With hyperfractionation, 90 (15%) patients had deviations (40 minor, 21 major acceptable, 29 major-unacceptable). As the assigned dose increased, the deviation rate increased (9.7% for 60.0 Gy vs. 20.8% for 79.2 Gy). Survivals for hyperfractionation patients with any deviations in treatment time were significantly shorter than those treated "per protocol" (p = 0.16): estimated 2- and 5-years rates were 24% and 10% versus 13% and 3%, respectively. Multivariate analyses showed the delay effect to be entirely in patients treated with 69.6 Gy or higher; there was also dependence upon the patients' prognosis. In patients with favorable prognosis (KPS 90-100, weight loss < or = 5%, no N3), the difference in survival was pronounced (33% and 15% vs. 14% and 0% at 2- and 5 years, respectively). Such differences were not found in patients with unfavorable prognostic factors. CONCLUSIONS: Interruptions delaying completion of planned radiation therapy were more frequent with higher total doses (> or = 69.6 Gy). Favorable patients (high KPS, little weight loss, < N3 nodal metastasis) had markedly adverse effects on long-term survival associated with delays to completion of the planned total dose. PMID- 8226141 TI - Neutron vs. photon radiation therapy for inoperable regional non-small cell lung cancer: results of a multicenter randomized trial. AB - PURPOSE: To determine, with a prospective, multicenter randomized study, whether fast neutron radiation therapy improves the outcome for patients with non-small cell lung cancer, as compared to conventional photon radiotherapy. METHODS AND MATERIALS: From September 1986 to March 1991, a total of 200 patients with inoperable regional non-small cell lung cancer were randomized to 20.4 Gy in 12 fractions with neutrons versus 66 Gy in 33 fractions with photons. Inoperable patients with Radiation Therapy Oncology Group Stages I, II, III, or IV(M0) disease, Karnofsky Performance Score > or = 70, and who had received no previous therapy for their non-small cell lung cancer were eligible for the study. Of the 200 patients randomized, a total of 193 patients, 99 on the neutron arm and 94 on the photon arm, were eligible for analysis. The two treatment groups were balanced with regards to prognostic factors. At the time of this analysis, the median at-risk follow-up was 33 months, with a minimum follow-up of 16 months. RESULTS: No difference in overall survival was observed; however, there was a statistically significant improvement in survival for patients with squamous cell histology (p = 0.02), and a trend toward improved survival for those with favorable prognostic factors (i.e., patients who were not T4, N3, and had no pleural effusion or weight loss > 5% from baseline) (p = 0.15), favoring the neutron-treated group. With the exception of skin and subcutaneous changes, acute and late toxicity was similar in both arms. CONCLUSION: In selected patients with inoperable regional non-small cell lung cancer (e.g., squamous cell histology, favorable prognostic factors), fast neutron irradiation provides a therapeutic benefit over conventional photon radiotherapy. PMID- 8226142 TI - Local control in medically inoperable lung cancer: an analysis of its importance in outcome and factors determining the probability of tumor eradication. AB - PURPOSE: For patients who are medically unable to tolerate a surgical resection for technically resectable non-small-cell lung carcinoma, radiation therapy is an acceptable alternative. We report on the effect of achieving local control of the primary tumor on survival end-points, and analyze factors that may influence local control. METHODS AND MATERIALS: We reviewed the records of 152 patients with medically inoperable non-small-cell lung carcinoma treated at our institutions. All patients had technically resectable lesions and no evidence of metastatic disease. Treatment was delivered using megavoltage irradiation to doses ranging from 45 to 75 Gy. RESULTS: For patients with tumors 3 cm or less, locally controlling the tumor significantly improved survival (p = .0371). Patients with T1 tumors had a higher probability of survival and disease-free survival than patients with larger tumors if the primary tumor was locally controlled, but this survival advantage disappeared if the tumor was not controlled. Overall, patients with smaller tumors had a lower incidence of distant spread, but this association was maintained only when the primary tumor was controlled (36 month risk of 10%, 23%, and 57% for tumors < 3 cm, 3-4.9 cm, 5 cm or greater, respectively, p = .0027). For patients whose tumors were not controlled, there was no significant difference in the risk of distant dissemination by tumor size. Higher radiation doses influenced local control and metastatic spread. We observed no influence of the initial field size in the risk of local control and in the probability of survival. CONCLUSION: Radical radiation therapy is an effective treatment for small (T1 or < 3 cm) tumors when treated to doses of 65 Gy or more, and should be offered as an alternative to surgery in elderly or infirm patients. New therapeutic strategies to improve the local control rate should be considered for larger tumors, through the use of hyperfractionated treatment, endobronchial "boost" irradiation, and sensitizing chemotherapy agents. PMID- 8226143 TI - Radiation therapy alone for stage I non-small cell lung cancer. AB - PURPOSE: This paper is a retrospective analysis of patients with clinical Stage I non-small cell carcinoma of the lung treated with definitive radiation therapy alone. The results of therapy, patterns of failure and the relationship of technical aspects of the delivery of radiotherapy to outcome are presented. MATERIALS AND METHODS: From 1980 through 1990, 53 patients with Stage I non-small cell lung cancer were treated with definitive radiation therapy alone at the Radiation Oncology Center of the Mallinckrodt Institute of Radiology and its affiliated hospitals. All patients had a pathologic diagnosis of non-small cell lung cancer and were not candidates for surgical resection because of either patient refusal (10 patients), poor performance status (5 patients), or premorbid medical problems (38 patients). The median age was 73 years. Histologic cell type included squamous (32), adenocarcinoma (11), large cell (4), and unclassified non small cell (6). Initial tumor size was < or = 3 cm in 23 patients, between 3 and 5 cm in 13 patients and > or = 5 cm in 17 patients. Diagnostic staging varied during the study period. All patients had chest X-rays and computed tomography scans of the chest. A majority had liver and bone scans, but only four underwent mediastinoscopy. The radiation therapy was of megavoltage energy in all patients, with a median primary prescription tumor dose of 63.2 Gy. Survival was measured from the date radiation therapy was initiated. RESULTS: The actuarial overall survival rate for the entire group was 19% at 3 years and 6% at 5 years, with a median survival time of 20.9 months. Of the 49 deaths, 35 died of lung cancer; 13 died of intercurrent illness, and one died of pancreatic cancer, which made the actuarial cause-specific survival 33% at 3 years and 13% at 5 years. The actuarial 3-year disease-free survival was 33%. Local primary tumor progression occurred in 22 patients, resulting in a 51% 3-year actuarial freedom from local progression. An additional four patients failed in regional lymph nodes that were included in the original treatment portals. Multivariate analysis found only T stage to be associated with overall survival (p = .02). However multivariate analysis showed age as a prognostic factor to be approaching statistical significance (p = .07). Patients under 70 years of age showed an increased survival rate compared to patients over 70 years. Radiation therapy doses > or = 65 Gy appeared to result in a decreased proportion of patients dying of lung cancer with no apparent increase in either acute or long-term complication rates. CONCLUSION: Results of definitive radiation therapy for inoperable Stage I non small cell lung remain inferior to surgical therapy. Potential methods to improve local control with radiotherapy are discussed. PMID- 8226144 TI - Postoperative radiotherapy after pneumonectomy: impact of modern treatment facilities. AB - PURPOSE: The present study was undertaken to see how modern treatment facilities, computed tomography (CT)-based treatment planning and linear accelerator, have modified the results of postoperative irradiation after a pneumonectomy for lung cancer. METHODS AND MATERIALS: Between 1970-1985, 103 patients were treated in our department after a pneumonectomy: 50 patients with a T1T2N0 tumor and 53 patients with a T3, N1 or N2 tumor. Three groups were considered: 27 patients had only surgical resection, 51 patients were irradiated postoperatively with a Co60 source, and 25 patients were treated using those modern facilities. RESULTS: The 5-year survival varies from 4% to 31% according to the tumor extent but also to the radiation technique. Patients treated with a Co60 source had a dismal 5-year survival rate (8%) whereas patients treated with the modern facilities had a 5 year survival rate of 30% similar to the 31% of the control surgical group including less advanced tumors. CONCLUSION: Linear accelerator and computed tomography-based treatment planning improved the accuracy of postoperative thoracic irradiation and allow to deliver high doses to the mediastinum even after a pneumonectomy. PMID- 8226145 TI - Inoperable endobronchial obstructing lung cancer treated with combined endobronchial and external beam irradiation: a dosimetric analysis. AB - PURPOSE: This study is a dosimetric review of our experience incorporating both external beam radiotherapy and endobronchial brachytherapy in the treatment of inoperable endobronchial obstructing lung cancer. METHODS AND MATERIALS: Sixty five patients with symptomatic inoperable endobronchial obstructing lung cancer received combined external beam radiotherapy and temporary endobronchial iridium (Ir)-192 implants between February 1987 and May 1991. External beam irradiation doses ranged from 55 to 66 Gy. Patients were given 2 to 4 temporary Ir-192 endobronchial implants at the site of endobronchial obstruction. Individual implant doses ranged from 2.7 to 10 Gy. Total implant doses ranged from 6 to 35 Gy. Total treatment doses ranged from 61 to 96.6 Gy. RESULTS: Sixty-three percent of patients had a complete local endobronchial response to treatment and 23% had a partial response. Forty-seven percent of patients receiving a total dose of less than 70 Gy experienced a complete local endobronchial response to treatment while 83% of patients receiving a total dose of 85 Gy or greater experienced a complete local endobronchial response to treatment. Sixty-six percent experienced an improvement in their performance status using an Eastern Cooperative Oncology Group performance scale. Survival from the end of treatment was 38% at 1 year and 23% at 2 years, with the median survival at 8 months. Complications occurred in 9/65 (14%) of patients; 0% in patients receiving a total dose of less than 70 Gy, 10% in patients receiving a total dose of 70 to 84.99 Gy, and 24% of patients receiving a total dose of 85 Gy or higher. CONCLUSION: The data from this study again confirms the feasibility of this form of treatment for inoperable endobronchial obstructing lung cancer. In addition, there appears to be a dose response for both local response and complications. PMID- 8226146 TI - The influence of field size and other treatment factors on pulmonary toxicity following hyperfractionated irradiation for inoperable non-small cell lung cancer (NSCLC)--analysis of a Radiation Therapy Oncology Group (RTOG) protocol. AB - PURPOSE: The risk of pulmonary toxicity, observed in an Radiation Therapy Oncology Group Phase I/II randomized dose escalation trial of hyperfractionated irradiation for nonsmall cell lung cancer, was analyzed with regard to custom vs. hand blocking and compliance to protocol specified treatment field parameters. MATERIALS AND METHODS: There were 832 evaluable cases analyzed. The protocol required field margins 2 cm beyond primary tumor and involved nodes. In 674, the field margins were considered "per protocol" or as having minor protocol variations. In 94, margins exceeded protocol specification ("excessive margin" group). In this group, the area (cm2) of the effective (blocked) field and the portion including lung was measured from simulator films with a computer scanning device. Based on size and location of the primary and nodal disease, "per protocol" fields were constructed and the area (cm2) of lung included beyond these margins was estimated. Patients from both groups who received less than 30 Gy to normal lung were excluded from analysis of pulmonary toxicity. RESULTS: Grade 1 acute lung toxicity was higher (p = .009) in the "excessive margin" group compared to the "per protocol" group, whereas late lung toxicity was not significantly different (p = .94). The risk of Grade 2 or greater acute toxicity increased as area of excess irradiated lung increased. Overall lung toxicity, defined as the greater of either acute or late toxicity, was evaluated by multivariate analysis, in relation to assigned dose, effective treated field area, and type of shielding. Overall maximum lung toxicity (> or = Grade 2) was significantly greater in the "excessive margin" group, when lung treated beyond protocol margins exceeded an area of 35 cm2, than in the "per protocol" group, but only when the effective treated field size was > or = 180 cm2 (68% vs. 37%; p = .02). This effect was independent of assigned total dose or type of shielding. CONCLUSION: For nonsmall cell lung cancer treated with hyperfractionated irradiation, the risk of overall pulmonary toxicity was increased for patients treated with field sizes in excess of protocol specified margins of tumor coverage in comparison to patients treated with protocol specified margins. This effect was seen only when the area of lung treated beyond protocol margins exceeded 35 cm2 and when the overall field size was below 180 cm2. PMID- 8226147 TI - Breast conservation therapy for early stage breast carcinoma with outstanding 10 year locoregional control rates: a case for aggressive therapy to the tumor bearing quadrant. AB - PURPOSE: Between 1982 and 1988 233 American Joint Committee on Cancer Stage I and II invasive breast carcinomas were prospectively treated in 225 women with conservative tumor excision, careful assessment of histopathological margins, and dose-adjusted irradiation to maximum doses of 70 Gy to the tumor bearing quadrant of the breast. METHODS AND MATERIALS: The pathological stages at presentation were T1N0 and T1N1 in 57% and 13% and T2N0 and T2N1 in 19% and 10% of the patients, respectively. All patients were irradiated according to a policy that, beyond the 50 Gy to the whole breast and draining lymphatics, the tumor-bearing quadrant was boosted in adjustment to the histopathological margin. Normal tissue margins of < 2 mm were considered positive, margins 2-5 mm close, and margins > 5 mm negative and were boosted with 20, 15, and 10 Gy, respectively. Patients in whom the margin could not be assessed were re-excised or boosted to 20 Gy. Re excisions with no residual carcinoma were not boosted. Most patients boosted to 20 Gy to the tumor-bearing quadrant received interstitial 192-Ir implantations. RESULTS: The actuarial local control rates in the treated breast were 97.5% at 10 years with three recurrences having occurred at a median of 4.5 years after completion of radiotherapy. An additional two patients failed regionally outside the irradiation portals. The overall and disease-free survival of the whole group is 87.5% and 77%, respectively. CONCLUSION: The approach to breast conservation therapy followed in this study has resulted in outstanding local control rates and suggests that there may be a subset of patients that could be irradiated to the tumor bearing quadrant only. PMID- 8226148 TI - Breast-conserving surgery and definitive radiation: a comparison between quadrantectomy and local excision with special focus on local-regional control and cosmesis. AB - PURPOSE: Breast-conserving surgery and definitive radiation as an alternative to mastectomy is a well-accepted practice. However, there is limited information addressing the extent of surgical resection. The purpose of this study is to compare the outcome of patients treated with local excision or quadrantectomy, followed by definitive radiation with particular emphasis on local-regional control and cosmetic results. METHODS AND MATERIALS: Between 1978 and 1989, 425 patients with Stage I and II breast cancer underwent conservative surgery followed by definitive radiation. Fifty-four patients had a local excision and 371 had a quadrantectomy. Median follow-up was 42 months. Axillary dissection (levels I/II) was performed in 317 patients, and of these 126 patients had positive axillary lymph nodes. Radiation consisted of 4500-5000 cGy to the breast with Co60 or 4 MV photons, plus a boost to the tumor site (356/425 patients) for a total dose of 6000-6500 cGy. Treatment of the regional lymph nodes was given to patients with undissected or inadequately dissected axillas and usually to patients with multiple positive lymph nodes. Of the patients with positive lymph nodes, 46% received systemic chemotherapy. RESULTS: The 5-year actuarial freedom from local-regional recurrence rates for patients treated with local excision and quadrantectomy followed by definitive radiation were 92% and 93%, respectively (p = 0.7). The 5-year actuarial survival rates for local excision and quadrantectomy were as follows: overall (83% and 82%; p = 0.7), cause-specific disease-free (74% and 71%; p = 0.9), and distant disease-free (82% and 76%, p = 0.4). Estimated 10 year results are also presented. Cosmetic analysis required a minimum follow-up of 5 years. In the local excision, 77% of the patients had excellent-good result, compared to 53% following quadrantectomy (p = 0.03). Excluding patients who received chemotherapy, the excellent-good scores were 76% and 57%, respectively (p = 0.1). The most unfavorable cosmetic results were associated with quadrantectomy followed by radiation with boost dose and chemotherapy, excellent good in 22%. CONCLUSION: Conservative surgery consisting of local excision or quadrantectomy resulted in comparable local-regional control, overall, disease free and distant disease-free survival, although cosmetic results were superior in the local excision group. PMID- 8226149 TI - The use of iodine-125 seeds as a substitute for iridium-192 seeds in temporary interstitial breast implants. AB - PURPOSE: We have previously reported that the use of iodine-125 seeds in temporary plastic tube interstitial implants may be more advantageous than iridium-192 seeds due to less patient and personnel radiation exposure, reduced shielding requirements, and significant dosimetric advantages. The impact of this isotope on the rate of local control and cosmetic outcome in patients with early stage breast cancer treated with interstitial implants for their irradiation "boost" remains to be defined. METHODS AND MATERIALS: We reviewed the treatment outcome of 402 consecutive cases of Stage I and II breast cancer undergoing breast conserving therapy between 1/1/80 and 12/31/87. All patients underwent excisional biopsy and received 45-50 Gy to the entire breast followed by a boost to the tumor bed using either electrons (104 patients), photons (15 patients), or an interstitial implant with either iridium-192 (197 patients) or iodine-125 (86 patients) to at least 60 Gy. Iodine-125 implants were primarily performed in patients with significant risk factors for local recurrence (71%) or in patients with large breasts (17%). Local tumor control and cosmetic outcome were assessed and contrasted between patients boosted with each modality. RESULTS: With a median follow-up of 59.3 months, 18 (4.5%) patients developed a recurrence of the tumor in the treated breast (4.4% 5-year actuarial rate). No statistically significant differences in the 5-year actuarial rate of local recurrence were noted between patients boosted with either iodine-125 (3.0%), iridium-192 (3.8%), electrons (5.4%), or photons (0%). Likewise, no significant differences in the percentage of patients obtaining good/excellent cosmetic results were noted between iodine-125 (93%), iridium-192 (88%), electrons (90%), or photons (82%). CONCLUSION: We conclude that patients with Stage I and II breast cancer undergoing breast conserving therapy and judged to be candidates for boosts with interstitial implants can be effectively treated with iodine-125 seeds. Use of this isotope results in less patient and personnel irradiation exposure and a better dose distribution than iridium-192, since dose optimization can be routinely employed. Overall, local control and cosmetic outcome have been excellent and are similar to either iridium-192, electrons, or photons. PMID- 8226150 TI - The role of mastectomy in patients with stage I-II breast cancer presenting with gross multifocal or multicentric disease or diffuse microcalcifications. AB - PURPOSE: Women with Stage I-II invasive breast cancer who present with gross multicentric disease or diffuse microcalcifications have a significant risk of breast recurrence when treated with conservative surgery and radiation. The purpose of this report is to present the results of mastectomy in this group of patients. METHODS AND MATERIALS: Between 1982 and 1989, 88 patients with clinical Stage I-II breast cancer who presented with clinical and mammographic evidence of gross multicentric disease or diffuse microcalcifications underwent modified radical mastectomy. Median followup was 4 years for the 57 patients with gross multicentric disease and 5.6 years for 31 patients with diffuse microcalcifications. At the time of mastectomy, 42% of patients were found to have positive axillary nodes. Following mastectomy, 15 patients received post mastectomy radiation and 35 patients received adjuvant systemic chemotherapy. RESULTS: When compared to a group of 1295 patients with unifocal, Stage I-II breast cancer, treated with conservative surgery and radiation during the same time period, patients with gross multicentric disease and diffuse microcalcifications had a significantly higher incidence of > or = 4 positive nodes, patients with gross multicentric disease had a lower incidence of positive resection margins following mastectomy and patients with diffuse microcalcifications were younger. The 5-year actuarial risk of an isolated local regional recurrence was 8% for patients with gross multicentric disease or diffuse microcalcifications and 7% for patients with unifocal disease. Patients with gross multicentric disease or diffuse microcalcifications and > or = 4 positive axillary nodes who did not receive post mastectomy radiation had an increased risk for local regional recurrence. There were no significant differences in the 5-year actuarial overall or relapse-free survival (88% and 73% gross multicentric disease, 97% and 86% diffuse microcalcifications and 90% and 79% unifocal disease), freedom from distant metastasis (76% gross multicentric disease, 90% diffuse microcalcifications, 86% unifocal disease) or incidence of contralateral breast cancer (10% gross multicentric disease, 13% diffuse microcalcifications, 8% unifocal disease) among the three groups. CONCLUSION: The present study demonstrates no increased risk of local-regional recurrence in patients with gross multicentric disease or diffuse microcalcifications undergoing mastectomy in contrast to the increased risk of breast recurrence in patients with gross multicentric disease undergoing conservative surgery and radiation. Indications for post mastectomy radiation include > or = 4 positive nodes or close or positive surgical margins. Despite a significantly higher incidence of > or = 4 positive nodes, patients with gross multicentric disease and diffuse microcalcifications have a 5-year actuarial overall and relapse-free survival comparable to a group of patients with unifocal disease treated with conservative surgery and radiation. PMID- 8226151 TI - Local recurrence versus new primary: clinical analysis of 82 breast relapses and potential applications for genetic fingerprinting. AB - PURPOSE: The purpose of this study was to perform a detailed clinical pathological analysis of breast relapses in patients treated with conservative surgery and radiation therapy in an effort to classify those relapses as true local recurrences or second primary tumors, and to assess the prognostic and therapeutic implications of such a classification system. METHODS AND MATERIALS: Of 990 patients treated with conservative surgery and radiation therapy at our facilities prior to December 1987, 82 patients have experienced a relapse in the conservatively treated breast as the primary site of failure. Patients were classified as having new primary tumors if they fulfilled any one of the following criteria: a) breast relapse occurring at a site distinctly removed from the original tumor; b) histology of the breast relapse compared with the original tumor consistent with a new primary; or c) DNA flow cytometry converting from an aneuploid primary to a diploid relapse. RESULTS: As of 2/92, with a median follow up of 5.4 years from the time of breast relapse, the overall 5-year survival rate following breast relapse was 55%. Forty-seven patients were classified as true recurrences and 33 patients were classified as new primaries. Patients classified as true recurrences had a shorter median time to breast relapse than patients classified as new primaries (3.16 years vs. 5.42 years, p < .05) and an inferior post breast recurrence survival rate compared to patients classified as new primaries (36% vs. 89%, p < .05). Residual disease outside of the recurrent tumor bed was also noted to be more frequent in patients classified as true recurrences compared to patients classified as new primaries (48% vs. 16%, p < .05). CONCLUSION: Based on the clinical and pathological criteria outlined, it appears that a significant portion of patients experiencing a relapse in the conservatively treated breast may have new primary tumors as opposed to true local relapses. Distinction between a true recurrence and a new primary tumor may have significant prognostic implications. Uncertainties associated with the clinical and pathological criteria are presented and further investigations with genetic fingerprinting techniques to establish the clonality of breast relapses are presented and discussed. PMID- 8226152 TI - Primary radiation therapy for endometrial carcinoma: a case controlled study. AB - PURPOSE: Primary radiation therapy is generally considered inferior to a surgical approach for patients with endometrial carcinoma and is reserved for patients with a high operative risk. These patients are usually elderly, have multiple medical problems and frequently die of intercurrent disease. To evaluate the efficacy of primary radiation therapy a case controlled analysis comparing corrected survival of patients treated with primary radiation to patients treated with surgical therapy with or without radiation therapy was performed. METHODS AND MATERIALS: Sixty-four patients treated with primary radiation therapy were retrospectively studied. A Kaplan-Meier product limit survival analysis was used to estimate survival among patients treated with primary radiation therapy. A case control study matched by clinical stage, tumor grade, and time of diagnosis was performed. The Mantel-Cox statistic was used to evaluated the equality of the survival curves. RESULTS: Primary radiation therapy was used to treat 9.0% of the patients with endometrial carcinoma during the study period. Cardiovascular disease, diabetes, age greater than 80 and morbid obesity were the most common indications. Ninety percent of patients had either Stage I or II disease. Forty eight of the 64 patients (75%) completed treatment which included both teletherapy and brachytherapy. Ten patients received brachytherapy only. Twelve complications, both acute and chronic, occurred in eleven patients (17%). Intercurrent disease accounted for 13 of the 36 (36%) of the deaths. Clinical stage of disease and histologic grade of the tumor were significant predictors of survival, p = 0.0001 and p = 0.013, respectively. The case controlled study of Stage I and II patients treated by primary radiation therapy matched to surgically treated controls showed no statistical difference in survival. Dilatation and curettage after the completion of radiation therapy was predictive of local control, p = 0.003. CONCLUSION: Although surgery followed by tailored radiation therapy has become widely accepted therapy for Stage I and II endometrial carcinoma, even in patients who are a poor operative risk, the survival with primary radiation therapy is not statistically different. PMID- 8226153 TI - Analysis of prognostic factors in localized gastric lymphoma: the importance of bulk of disease. AB - PURPOSE: To identify prognostic factors in localized gastric lymphoma patients for optimal therapy selection. METHODS AND MATERIALS: From 1974 to 1990, 77 patients with localized gastric lymphoma (38 Stage IE and 39 Stage IIE) were treated with radiation therapy, chemotherapy, surgery, or a combination. Univariate and multivariate local control and survival analyses were performed on possible prognostic factors, such as patient age, gender, histologic subtype, stage, tumor size, depth of penetration, multicentricity, and treatment modality. RESULTS: At 5 years, the relapse-free survival was 52%; 74% of the relapses occurred at local sites. Smaller tumor size was most strongly associated with local control (p = .001) and relapse-free survival (p < .001). Patients with tumor sizes < or = 5 cm had relapse-free survival of 87%, compared with 41% and 15% for those with tumor sizes of 5.1 cm to 10 cm and > 10 cm, respectively. The 47 patients who received combined-modality therapy had a relapse-free survival of 65%, compared with 24% for the 30 who received single-modality therapy (p < .01). Although patient age, stage, depth of penetration, and resective surgery affected the above endpoints, these factors were not independent predictors of outcome. Analysis of treatment subgroups showed that surgical resection combined with postoperative irradiation was associated with highest local control (p = .002) and the best relapse-free survival (p = .004), when compared with other treatment modalities. In 27 patients with tumor sizes < or = 5 cm, comparison of the 15 patients who had surgery with the 12 who did not failed to reveal a local control benefit from the addition of surgery. CONCLUSION: These data demonstrate that tumor bulk is an important prognostic determinant of local control and relapse free survival in localized gastric lymphoma patients. Stage IE and IIE lymphoma of the stomach can be selectively treated with primary radiation, but surgical resection may be necessary for large tumors (> 5 cm), followed by adjuvant radiation. PMID- 8226154 TI - Non-Hodgkin's lymphoma of the thyroid gland: prognostic factors and treatment outcome. The Princess Margaret Hospital Lymphoma Group. AB - PURPOSE: Non-Hodgkin's lymphoma presenting in the thyroid gland is uncommon. A review of the Princess Margaret Hospital experience was performed to assess treatment outcome and prognostic factors in this rare extranodal presentation of localized lymphoma. METHODS AND MATERIALS: Fifty-two patients treated at the PMH between 1978 and 1986 were identified and their records reviewed retrospectively. Staging procedures revealed 16 patients with Stage I, 28 with Stage II, and eight with Stages III or IV disease. Five patients were treated on a protocol designed for anaplastic carcinoma of thyroid and they were excluded from detailed analysis. Of 39 patients with Stages I and II disease, 18 were treated with radiotherapy alone, three chemotherapy alone, and 18 combined modality therapy. Combined modality therapy was used mainly in patients with large tumor bulk. RESULTS: The overall 5-year actuarial survival and cause-specific survival were 56% and 64%, respectively. The overall relapse-free rate was 61% at 5 years. Among the 39 patients with Stages I and II disease, the 5-year actuarial survival, cause-specific survival, and relapse-free rate were 64%, 73%, and 66%, respectively. There were no significant differences in outcome between those treated with radiotherapy alone and those treated with combined modality therapy (cause-specific survival: p = 0.25, relapse: p = 0.06). A univariate analysis showed that the only variable to reach statistical significance was tumor bulk. Age was marginally significant while stage and histology were not statistically significant, possibly due to the fairly homogeneous distribution of patients in each of these variables. Patients with progression or relapse of lymphoma after initial treatment frequently died of disease. Isolated gastrointestinal relapses occurred in three cases, representing 27% of all relapses. CONCLUSION: Based on the above results, we recognize that the majority of patients with localized thyroid lymphoma require combined modality therapy and we recommend radiotherapy alone only for a small, select group of patients with Stage I disease and small tumor bulk. PMID- 8226155 TI - Timing and sequence of hyperthermia in fractionated radiotherapy of a murine fibrosarcoma. AB - PURPOSE: This study investigated the effect of timing and sequence of hyperthermia on fractionated radiotherapy, since it has been shown that the heat increases the size of hypoxic cell fraction which could affect the effect of subsequent radiation doses. METHODS AND MATERIALS: Animal-tumors were early generation isotransplants of a spontaneous fibrosarcoma, FSa-II, in C3Hf/Sed mice. Tumor response was studied by tumor growth time and TCD50 (50% tumor control dose) assays. The tumor growth time is the time required for one-half of the treated tumors to reach 500 mm3 from the first treatment day. The TCD50 is a radiation dose to control one-half of the treated tumors for 120 days following treatments. One heat treatment at 43.5 degrees C for 45 min was given in a water bath in combination with fractionated doses independently (24 hr interval) or simultaneously (2 min interval). For the normal tissue study, the mouse foot was treated, and the acute foot reaction was scored daily and averaged. The late foot reaction was scored in animals used in the TCD50 assay that developed no recurrence for 120 days. The RD50(2.0) and RD50(5.0), or total radiation doses to induce an average score of 2.0 (complete epilation) and 5.0 (partial foot atrophy) in 50% of treated animals, were calculated. RESULTS: Thermal radiosensitization was most prominent when heat was combined simultaneously with the first or last radiation dose in both the tumor growth time and TCD50 assays. However, the thermal enhancement was greatest when heat was given either with the first or last radiation dose in the TCD50 assay; whereas it was greatest when heat was administered with the last radiation dose in the tumor growth time assay. Both acute and late skin reactions were significantly potentiated by heat administered 24 hr before the first radiation dose. CONCLUSION: A significant observation in this study was that, in both the tumor growth time and TCD50 assays, heat given independently or simultaneously did not result in any therapeutic gain compared to the radiation alone treatment. PMID- 8226156 TI - Combined Betaseron R (recombinant human interferon beta) and radiation for inoperable non-small cell lung cancer. AB - PURPOSE: Based on in vitro evidence of radiosensitization by Betaseron (beta IFN), a Phase I/II study was undertaken to determine toxicity and response using combined radiation (RT) and B-IFN in patients with unresectable Stage III and nonsmall cell lung cancer. METHODS AND MATERIALS: Varying doses of beta-IFN(10 to 90 x 10(6) IU) were administered IV immediately preceding RT on the first three days of weeks 1, 3, and 5. The RT dose was 1.8 Gy/day, 5 days/week for a total of 54 or 59.4 Gy. RESULTS: Thirty-nine patients were entered, 32 of whom were evaluable. The median follow-up time at time of analysis was 60 months. Responses were based on CT scan. The response rate for the total group was 81% with 44% achieving complete response. Seventy-eight percent of patients with complete response survived a minimum of 21 months. Twenty-six patients had Stage III A/B disease with a median tumor size of 6.5 cm. and median survival was 19.7 months. The 5-year actuarial survival for this group was 31%, with a plateau persisting after 3 years. There were no treatment related deaths nor any event of life threatening toxicity. Of eight patients surviving 3-5 years, no long-term toxicity has been observed. Karnofsky indices were 90-100 and respiratory symptoms were minimal. CONCLUSION: beta-IFN is well-tolerated. Response and survival rates are sufficiently encouraging to warrant further investigation in a randomized trial which has been accepted as an RTOG study awaiting drug availability. PMID- 8226157 TI - Preoperative concomitant cisplatin/VP16 and radiotherapy in stage III non-small cell lung cancer. AB - PURPOSE: To evaluate the therapeutic effectiveness of a combined chemoradiotherapy program, followed by surgery in selected cases, in Stage III non-small cell lung cancer. METHODS AND MATERIALS: Between August 1988 and February 1990, 43 patients Staged IIIa-b (UICC 1987, 58% IIIb) have been treated with concomitant chemotherapy (cisplatin 15 mg/m2 and VP16 75 mg/m2, 5 days a week on week 1 and 5) and radiotherapy (40 Gy split course, 2 Gy/day on week 1, 2, 5, and 6), followed by attempted curative thoracotomy or more cycles of full dose chemotherapy with the same two drugs. RESULTS: Planned chemoradiotherapy has been given to 91% of patients; 13/43 patients have been operated, with 12 complete resections and three (7%) pathological complete responses. Toxicity was significant, with two postoperative deaths and two fatal radiation pneumonitis. Crude progression-free survival rate is 21% at 30 months, with nine patients (21%) alive and free from progression at follow-up times ranging from 31 to 49 months. Subset survival analysis showed a possibly greater therapeutic effect for non-squamous histology as compared to squamous carcinoma. CONCLUSION: These results are encouraging in a cohort of patients with quite advanced disease (58% Stage IIIb). PMID- 8226158 TI - Radioresponsiveness of human glioma, sarcoma, and breast cancer spheroids depends on tumor differentiation. AB - PURPOSE: Differences in the intrinsic radiosensitivity within and between different tumor classes have been noticed for human tumor cell lines using the clonogenic assay. By far, most of the cell lines studied up to now were derived from poorly differentiated tumors. In this study, the influence of tumor differentiation on the radiation doses necessary to control 50% of small oxic spheroids (SCD50) was determined. Evidence of a distinct dependence of radioresponsiveness on tumor progression provides a background for an investigation of the underlying mechanisms. METHODS AND MATERIALS: Spheroids were aggregated from 1000-1500 cells in agarose coated 24 multi-well plates. Their diameters ranged from 156 to 405 microns, depending on the cell line. Spheroids were irradiated with graded 60Co single doses using spheroid control as end point and a minimum follow-up period of 3 months. RESULTS: Cell lines from three low grade gliomas and 10 malignant gliomas were studied in the spheroid control assay. The group mean SCD50 values were 6.1 +/- 1.6 Gy and 13.1 +/- 3.3 Gy, respectively. Four cell lines from grade 2 soft tissue sarcomas had a mean SCD50 value of 6.2 +/- 0.5 Gy and one undifferentiated sarcoma line of 11.0 Gy. Three well-differentiated breast cancer lines expressed the cell adhesion molecule E cadherin, had an epithelioid morphology in monolayer culture, were estrogen receptor positive or contact inhibited in multicellular spheroids. Two undifferentiated breast cancer lines had a fibroblastoid morphology and were marker negative. The mean SCD50 value of the former was 10.5 +/- 1.0 Gy while that of the undifferentiated lines was 14.8 +/- 2.8 Gy. Analysis of variance revealed a significant effect of the tumor type as well as the grade of dedifferentiation on the SCD50 after irradiation with one fraction or 2Gy/fraction. The surviving fractions at 2 Gy (SF2), obtained from the spheroid control rates after different fractionation schedules by approximation of the linear quadratic model assuming Poisson statistics were significantly dependent on tumor type (p = 0.001, ANOVA F-test) but not on tumor differentiation (p = 0.27). The alpha/beta ratios did not depend on tumor type (p = 0.08, ANOVA F Test) but significantly increased with the grade of tumor cell dedifferentiation (p = 0.03). CONCLUSION: The spheroid model is suitable for measuring the radioresponsiveness of differentiated cell lines with very low colony forming efficiencies. Tumor cell differentiation is an important factor for the radioresponsiveness and recovery capacity of human tumor cells. PMID- 8226159 TI - Cell cycle alterations, apoptosis, and response to low-dose-rate radioimmunotherapy in lymphoma cells. AB - PURPOSE: In an attempt to elucidate some aspects of the radiobiological basis of radioimmunotherapy, we have evaluated the in vitro cellular response patterns for malignant lymphoma cell lines exposed to high- and low-dose-rate radiation administered within the physiological context of antibody cell-surface binding. METHODS AND MATERIALS: We used two different malignant lymphoma cell lines, a Thy1.2+ murine T-lymphoma line called EL-4 and a CD20+ human B-lymphoma line called Raji. Cells were grown in suspension cultures and exposed to high-dose rate gamma radiation from an external 137Cs source or low-dose-rate beta radiation from DTPA-solubilized 90Y in solution. In some experiments, cells were pre-incubated with an excess of nonradioactive antibody in order to assess the effects of immunoglobulin surface binding during radiation exposure. Irradiated cells were evaluated for viability, cell-cycle changes, patterns of post radiation morphologic changes, and biochemical hallmarks of radiation-associated necrosis and programmed cell death. RESULTS: The EL-4 line was sensitive to both high-dose-rate and low-dose-rate irradiation, while the Raji showed efficient cell kill only after high-dose-rate irradiation. Studies of radiation-induced cell cycle changes demonstrated that both cell lines were efficiently blocked at the G2/M interface by high-dose-rate irradiation, with the Raji cells appearing somewhat more susceptible than the EL-4 cells to low-dose-rate radiation-induced G2/M block. Electron microscopy and DNA gel electrophoresis studies showed that a significant proportion of the EL-4 cells appeared to be dying by radiation induced programmed cell death (apoptosis) while the Raji cells appeared to be dying primarily by classical radiation-induced cellular necrosis. CONCLUSION: We propose that the unusual clinical responsiveness of some high and low grade lymphomas to modest doses of low-dose-rate radioimmunotherapy may be explained in part by the induction of apoptosis. The unusual dose-response characteristics observed in some experimental models of radiation-induced apoptosis may require a reappraisal of standard linear quadratic and alpha/beta algorithms used to predict target tissue cytoreduction after radioimmunotherapy. PMID- 8226160 TI - Ultrasonic measurements of breathing rate in rats and computer assisted analysis. AB - PURPOSE: An ultrasound breathing rate measurement technique and a computer analysis algorithm have been developed to reduce the amount of time needed to collect and analyze animal breathing rate data, as well as to improve the testing environment. The system is not airtight, therefore, acclimatization and collection time is not limited, and the technique makes use of a top loading apparatus to facilitate animal entry. METHODS AND MATERIALS: Breathing rate is measured using two ultrasound transducers housed directly above the rat thorax in the plexiglass jig. The breathing rate signal is stored and evaluated by computer. The ultrasound technique was tested using a loud speaker driven by a signal generator, over a range of 30 to 450 cycles/min. In addition, the ultrasonic breathing rate method was used to record the breathing rate response of Sprague Dawley rats, treated with graded single doses of radiation, over a period of 170 days. RESULTS: For the loud speaker tests, the measured frequency agreed with that of the input signal with a maximum deviation of 1%. For the animal irradiations, all breathing rate data were analyzed by both user and computer selection of regular breathing. The techniques gave the same results at the 95% confidence limit. Using the computer program to assess the traces, 240 breathing rates can be determined per hour, from previously measured data. CONCLUSION: A new technique for measuring breathing rate has been developed and enhances both the collection and analysis of data. PMID- 8226161 TI - The use of biphasic linear ramped pulsed field gel electrophoresis to quantify DNA damage based on fragment size distribution. AB - PURPOSE: The development of biphasic linear pulse ramping gel electrophoresis has permitted resolution of DNA fragments from 200 Kbp to 6 Mbp in a single gel. We used this technique to measure radiation-induced DNA damage based on fragment size. METHODS AND MATERIALS: Human colon cancer cells (HT29 and LS174T) and Chinese hamster ovary cells were embedded in agarose, deproteinized, irradiated with 5-80 Gy, and assessed for DNA double strand breakage using pulsed field gel electrophoresis. The frequency of DNA double strand breakage determined using a previously published method was compared to the breakage frequency calculated using the fragment size distribution. RESULTS: Both methods produced similar estimates for breakage frequency of approximately 5 x 10(-9) breaks Gy-1 bp-1. CONCLUSIONS: These findings suggest that biphasic linear pulse ramping gel electrophoresis can yield a quantitative estimate of DNA fragment distribution resulting from irradiation. The ability to quantify the distribution of DNA fragment sizes produced by irradiation should yield important information concerning the mechanisms of both DNA double strand break induction and repair. PMID- 8226162 TI - Interaction of the bioreductive drug SR 4233 and photodynamic therapy using photofrin in a mouse tumor model. AB - PURPOSE: Combining the bioreductive drug SR 4233 with interstitial photodynamic therapy to improve efficacy. METHODS AND MATERIALS: RIF1 tumors were implanted subcutaneously in mice and treated with interstitial photodynamic therapy. The bioreductive drug SR 4233 (a benzotriazine which exhibits preferential cell killing under hypoxic conditions) was combined with photodynamic therapy to exploit the induced hypoxia. SR 4233 was given to mice prior to or just after illumination. The effect of multiple SR 4233 injections given over the first 3 days after treatment was also evaluated. RESULTS: The results from experiments with a 24 hr interval between Photofrin and illumination showed that SR 4233 produced only a small additional growth delay compared with photodynamic therapy alone (light doses of 300 or 400 J/cm, combined with 6 x 15 mg/kg SR 4233). Some cures (6/60), however, were found in groups treated with 200 to 400 J/cm with SR 4233, whereas only two cures (2/77) occurred at light doses up to 400 J/cm after photodynamic therapy alone. Reducing the interval between Photofrin injection and illumination increased the number of cures in the combination group, although this was associated with a marked increase in toxicity. A small increase in cure rate was observed for the combination of photodynamic therapy (6 hr interval) and SR 4233, although this was not significant due to the limited number of mice that survived treatment. CONCLUSION: Only a limited effect of combining SR 4233 and interstitial photodynamic therapy was observed in this tumor model. A possible explanation could be the rapid conversion of SR 4233 into inactive metabolites. PMID- 8226163 TI - Primary soft tissue sarcomas of the breast: local-regional control with post operative radiotherapy. AB - PURPOSE: A retrospective analysis was undertaken to determine the efficacy of postoperative radiotherapy in patients with primary sarcoma of the breast. METHODS AND MATERIALS: Ten patients with high-grade nonmetastatic primary sarcoma of the breast were treated at the National Cancer Institute, NIH, between 1979 and 1989 with mastectomy and adjuvant radiotherapy. Chemotherapy was given to three patients as part of a randomized trial. RESULTS: With a median potential followup of 99 months postoperatively, seven patients remain alive and without evidence of disease 142, 119, 82, 48, 45, 28, and 19 months postoperatively. Three patients died of metastatic disease 7, 25, and 29 months, postoperatively. There were no local or regional failures. Actuarial 5-year disease free and overall survival were 68% and 66%, respectively. CONCLUSION: Sarcomas of the breast have a prognosis similar to that of extremity sarcomas. When adjuvant radiotherapy is used, excellent local control may be achieved. PMID- 8226164 TI - IR-192, low dose rate endobronchial brachytherapy in the treatment of malignant airway obstruction. AB - PURPOSE: To assess the value of low-dose-rate endobronchial brachytherapy in the treatment of malignant airway obstruction. METHODS AND MATERIALS: Between September 1986 and April 1989, 39 patients with malignant airway obstruction had 49 catheter placements for an afterloading, low-dose-rate Ir-192 endobronchial brachytherapy. A flexible fiberoptic bronchoscope with fluoroscopic guidance was used for positioning. Thirty-eight of 39 (97%) patients completed the prescribed treatments. Ninety-seven percent had received previous external radiation in doses ranging from 36-60 Gy. One patient had metastatic renal cell carcinoma; the remainder had recurrent lung cancer. Endobronchial laser treatments were given to three patients 2-3 weeks prior to endobronchial brachytherapy. All patients were followed until death. The median dose delivered in 48 of the 49 placements was 20 Gy at 1 cm. RESULTS: Follow-up bronchoscopy was performed in 28 (72%) of 39 patients. Of these, 13 (46%) had a complete response, 12 (43%) had a partial response, and 3 (17%) had a minor response. Dyspnea improved in 30 of 37 patients (82%); hemoptysis in 17 of 19 patients (89%); cough in 31 of 39 patients (79%); and postobstructive pneumonia in 21 of 23 patients (92%). The median survival for the entire group was 5 months (range 1-31 months). CONCLUSION: This technique is simple, well-tolerated and offered significant palliation. PMID- 8226165 TI - Independent collimator dosimetry for a dual photon energy linear accelerator. AB - PURPOSE: The independent collimator feature in medical linear accelerators can define radiation fields that are asymmetric with respect to the flattening filter and oblique to the incident surface. Prior to clinical implementation, it is necessary to evaluate the dosimetry of this non-standard treatment delivery technique. An investigation of the independent collimator dosimetry for 6 MV and 18 MV x-ray beams has been undertaken. METHODS AND MATERIALS: Dose to tissue in free space, percent depth dose and dose distribution were measured and compared to that for symmetric field collimation. RESULTS: The dosimetry results were consistent for both photon modes. Dose in free space with asymmetric collimation can be calculated from the corresponding symmetric field dose in free space to within 1.2 +/- 0.7% by applying an appropriate off-axis factor. Asymmetric field percent depth dose differs from symmetric field percent depth dose on average by 1.1 +/- 0.7% for 6 MV and by 0.7 +/- 0.5% for 18 MV for field sizes ranging from 5 x 5 to 20 x 20, centered 3 cm and 10 cm off-axis. The measured isodose curves demonstrate divergence effects and reduced doses (less than 3%) adjacent to the field edge closest to the flattening filter center. This dose asymmetry result is identical to that from secondary collimation. CONCLUSION: The methodology for clinical implementation of the independent collimator feature is straightforward. However, accurate representation of the isodose distributions by commercial radiotherapy treatment planning systems requires special dose calculation algorithms. PMID- 8226166 TI - Use of Veff and iso-NTCP in the implementation of dose escalation protocols. AB - PURPOSE: This report investigates the use of a normal tissue complication probability (NTCP) model, 3-D dose distributions, and a dose volume histogram reduction scheme in the design and implementation of dose escalation protocols for irradiation of sites that are primarily limited by the dose to a normal tissue which exhibits a strong volume effect (e.g., lung, liver). METHODS AND MATERIALS: Plots containing iso-NTCP contours are generated as a function of dose and partial volume using a parameterization of a NTCP description. Single step dose volume histograms are generated from 3-D dose distributions using the effective-volume (Veff) reduction scheme. In this scheme, the value of Veff for each dose volume histogram is independent of dose units (Gy, %). Thus, relative dose distributions (%) may be used to segregate patients by Veff into bins containing different ranges of Veff values before the assignment of prescription doses (Gy). The doses for each bin of Veff values can then be independently escalated between estimated complication levels (iso-NTCP contours). RESULTS AND CONCLUSION: Given that for the site under study, an investigator believes that the NTCP parameterization and the Veff methodology at least describe the general trend of clinical expectations, the concepts discussed allow the use of patient specific 3-D dose/volume information in the design and implementation of dose escalation studies. The result is a scheme with which useful prospective tolerance data may be systematically obtained for testing the different NTCP parameterizations and models. PMID- 8226167 TI - Combining multileaf fields to modulate fluence distributions. AB - PURPOSE: A method for modulating beam fluence from a linear accelerator is discussed. The beam modulation is accomplished remotely using a multileaf collimator and does not require entering the treatment room. METHODS AND MATERIALS: The multileaf collimator is used to define a series of field shapes that are superimposed at a fixed gantry angle to produce any desired fluence pattern. A heuristic technique for deriving the field shapes and corresponding monitor unit settings is described. The technique has been tested on randomly generated fluence distributions and on distributions with a limited number of peaks and valleys. The second type of distribution more closely simulates fluence patterns obtained with dose optimization software. Estimates of the time required to use this approach to treat a four-field plan are given and compared to the technique of placing a physical compensator in each beam. RESULTS: It has been demonstrated that complex fluence patterns within a 15 x 15 cm2 field can be achieved with less than 20 fields. Estimates show that this technique is faster than entering the treatment room to change physical compensators. Some limitations of the method are discussed. CONCLUSION: Optimized distributions that conform the dose to irregularly shaped target volumes that wrap around critical structures are possible using superimposed multileaf fields. A method for defining the field shapes is presented. PMID- 8226168 TI - The use of on-line image verification to estimate the variation in radiation therapy dose delivery. AB - PURPOSE: On-line radiotherapy imaging systems provide data that allow us to study the geometric nature of treatment variation. It is more clinically relevant to examine the resultant dosimetric variation. In this work, daily beam position as recorded by the on-line images is used to recalculate the treatment plan to show the effect geometric variation has on dose. METHODS AND MATERIALS: Daily 6 MV or 18 MV x-ray portal images were acquired using a fiberoptic on-line imaging system for 12 patients with cancers in the head and neck, thoracic, and pelvic regions. Each daily on-line portal image was aligned with the prescription simulation image using a template of anatomical structures defined on the latter. The outline of the actual block position was then superimposed on the prescription image. Daily block positions were cumulated to give a summary image represented by the block overlap isofrequency distribution. The summary data were used to analyze the amount of genometric variation relative to the prescription boundary on a histogram distribution plot. Treatment plans were recalculated by considering each aligned portal image as an individual beam. RESULTS: On-Line Image Verification (OLIV) data can differentiate between systematic and random errors in a course of daily radiation therapy. The data emphasize that the type and magnitude of patient set-up errors are unique for individual patients and different clinical situations. Head and neck sites had the least random variation (average 0-100% block overlap isofrequency distribution width = 7 mm) compared to thoracic (average 0-100% block overlap isofrequency distribution width = 12 mm) or pelvic sites (average 0-100% block overlap isofrequency distribution width = 14 mm). When treatment delivery is analyzed case by case, systematic as well as random errors are represented. When the data are pooled by anatomical site, individuality of variations is lost and variation appears random. Recalculated plans demonstrated dosimetric deviations from the original plans. The differences between the two dosimetric distributions were emphasized using a technique of plan subtraction. This allowed quick identification of relative "hot and cold spots" in the recalculated plans. The magnitude and clinical significance of dosimetric variation was unique for each patient. CONCLUSIONS: OLIV data are used to study geometric uncertainties because of the unique nature for individual patients. Dose recalculation is helpful to illustrate the dosimetric consequences of set-up errors. PMID- 8226169 TI - Calculation and measurement of the dose to points outside the primary beam for CO 60 gamma radiation. AB - PURPOSE: In radiation therapy one sometimes needs to estimate the dose to points in the body outside the primary beam. Therefore a generalized model is developed to calculate this dose with reasonable accuracy. METHODS AND MATERIALS: Measurements were made for a cobalt beam to determine separately the contribution of leakage radiation, radiation scattered from the collimator, scattered from the floor and radiation scattered inside the patient. RESULTS: The radiation scattered in the patient shows a strong dependence on field size and distance to the beam axis and is predominant only at short distances. The radiation scattered from the collimator also depends strongly on distance and field size and is more important than the leakage radiation. With appropriate factors, correcting for patient dimensions and field shape, the total dose outside the primary beam can be calculated with an accuracy better than +/- 30%. The results are in accordance with published data. CONCLUSION: Using the measured data it is possible to calculate the dose at any point of the body outside the primary beam for Co-60 gamma radiation. The accuracy is considered to be adequate for risk assessment. Gonadal dose, Radiation therapy, Risk assessment. PMID- 8226170 TI - Anatomic variation of gynecologic brachytherapy prescription points. AB - PURPOSE: The purpose of this report is to evaluate the geometric movement (relative to the bony pelvis) of fixed brachytherapy reference points during the time interval of the first and second gynecologic intracavitary implant. METHODS AND MATERIALS: The radiation therapy records of 40 consecutive patients with all stages of carcinoma of the cervix treated at the Radiation Oncology Center, Mallinckrodt Institute of Radiology, from January 1991 through December 1991 were reviewed. All patients received external beam irradiation and two intracavitary implants with Fletcher-Suit tandem and ovoids. Prescription points, per ICRU #38, were used: point A, B, and P, bladder, and rectum. Comparison of the location of points for the first and second implants (2 weeks time interval) was performed using the bony pelvis as reference anatomy and calculating the 3-dimensional spherical coordinates from a common origin for both implants. RESULTS: Analysis of the movement of the second implant relative to the first revealed that the x, y, and z coordinates for the various points shifted from 0 to 6 mm in a single plain. Vector analysis showed that the magnitude and direction of the average shift was on the order of 1.0 to 1.5 cm with displacement posteriorly and inferiorly. The effect of this movement on dose rates to the various points can result in dose rate differences up to 35%. A regression analysis was performed to identify factors affecting this movement. The time interval from the first to the second implant was the factor which correlated best with the movement. CONCLUSION: There is movement of the absolute position of ICRU #38 reference points between the first and second intracavitary implants. This movement results in significant differences in absolute dose rates to these reference points in the two implants. The clinical significance of this movement relative to dose to the tumor is unknown. PMID- 8226171 TI - Preservation of parotid function after external beam irradiation in head and neck cancer patients: a feasibility study using 3-dimensional treatment planning. AB - PURPOSE: Radiation-induced xerostomia is a frequent complication and major cause of morbidity in head and neck cancer patients. The severity of xerostomia is related to radiation dose and the amount of parotid tissue included in the irradiated volume. To reduce this side-effect and preserve salivary function, we have evaluated the use of 3-dimensional (3-D) treatment planning to spare the contralateral parotid gland in twelve patients undergoing radiation therapy for head and neck cancers. METHODS AND MATERIALS: In each case, beam's eye view displays were used to design beam and blocking arrangements that excluded the contralateral parotid. Ten patients were treated with 2 nonopposing oblique fields in the axial and non-axial plane while two patients required a non-axial, non-coplanar 3-field arrangement. These 3-D treatment plans were also compared with conventional 2-dimensional (2-D) plans. The 2-dimensional plans were designed independently of the 3-D treatment planning information using the orthogonal radiographs and hard copies of the computed tomography scans. RESULTS: An average of 1.8% (range, 0-7%) of the target volume was underdosed with the 95% isodose level for the 3-D plans compared with 18.8% (range, 2.0-36.6%) for the 2 D plans. This was due to improved identification of the target volumes and better design of blocked fields with beam's eye view treatment planning. Furthermore, the mean dose to the opposite parotid was 3.9 Gy for the 3-D plans vs 28.9 Gy for the conventional plans. With a minimum follow-up of 4 months, only 2 of 12 patients have complained of a dry mouth. CONCLUSION: These encouraging results suggest that this approach is feasible in many cases. 3-D treatment planning may allow the use of parotid sparing techniques in patients who otherwise would not have been considered candidates using conventional radiotherapy techniques. PMID- 8226173 TI - Is a nihilist approach to lung cancer still justified? PMID- 8226172 TI - Radiation and simultaneous cisplatin in non-small cell lung cancer. AB - PURPOSE: Phase III non-small cell lung cancer trials comparing radiation and simultaneous single agent cisplatin-radiation, as well as, Phase II trials of cisplatin containing combination regimens and concurrent thoracic radiation used as preoperative or as definitive therapy in stage III non-small cell lung cancer are reviewed. METHODS AND MATERIALS: The prognostic significance of the new international staging system with respect to clinical Stage III disease is described and discussed in this review because it has important implications for clinical trials. The results of four randomized Phase III trials and one Phase II trial which evaluated radiation therapy and single agent cisplatin are reviewed. The data from studies of combination chemotherapy and concurrent thoracic radiation observed in two consecutive Rush University Phase II trials and in a randomized Phase II Mayo Clinic trial are described. Eight phase two studies in which thoracic radiation and simultaneous cisplatin containing combination chemotherapy were given as preoperative treatment are compared. RESULTS: Studies evaluating the prognostic significance of the new staging system (IIIa vs IIIb) have shown conflicting results. In Rush University trials there has been a significant difference for IIIa versus IIIb and in particular the tumors which are invading the mediastinum or chest wall without obvious mediastinal lymph node metastases appear to have the best prognosis. Similarly randomized trials evaluating curative doses of thoracic radiation therapy with or without current single agent cisplatin have shown contradictory results. One of the four randomized trials have shown superior survival with patients treated with radiation and simultaneous daily cisplatin. Toxicity with cisplatin combination chemotherapy regimens and split course radiation has been acceptable. In Phase II non-surgical trials preoperative treatment consisting of cisplatin containing combination regimens given simultaneously with thoracic radiation have shown that this type of combined modality therapy is feasible and that the rates of resectability appear to be higher than would be expected with surgery alone. Survival results from six of these studies appear to be superior to results reported for radiation or surgery alone. CONCLUSION: Additional data are needed to determine the prognostic significance of the new staging system for clinical Stage III non-small cell lung cancer patients. Similarly, additional Phase III trials will be required to determine the role of thoracic radiation and concurrent single agent cisplatin, as well as, concurrent cisplatin combination regimens. Treatment with preoperative radiation and concurrent cisplatin containing combination therapies is feasible and relatively safe. Phase III trials are needed to determine the impact of neoadjuvant chemoradiation therapy and surgery in Stage III patients. PMID- 8226174 TI - When is more better? PMID- 8226175 TI - Can interferon beta make the radiation therapy team for treatment of non-small cell lung cancer? PMID- 8226176 TI - Response to Drs. Brada and Thomas. PMID- 8226177 TI - A systems therapist looks at transference: widening the lens. PMID- 8226178 TI - The phenomenology of the self and its objects in waking and dreaming: implications for a model of dreaming. AB - The current psychoanalytic emphasis on self psychology allows attention to be drawn to the nature of the self, internal and external object representations, and possibilities involving the self in relationship to aspects of self and objects in dreams. An emphasis on the self, applied to dreams, appears to fit well with current concepts of REM sleep that involve this sleep stage's importance for the individual in learning, adaptation, and assimilation of emotionally meaningful information. Examples have been provided to indicate possible contributions of self psychological perspectives to an understanding of dreams. A model describing changes in the self and its objects in dream mentation compared to the self and its objects in waking mentation has been described. The model suggests that the self of the dream relates to separated off, changed, or newly cathected attitudes, beliefs, and object representations in a manner that describes the state of the individual and possibilities for change available to the individual. This model, having historical roots in Jungian psychology, adaptive/learning theories of REM sleep, and current Kohutian initiated self state dream theory, provides an opportunity to review with the dreamer the relationships of self and objects as these representations are concerned with problematic developmental or emotional situations. PMID- 8226179 TI - Unlocking incest memories: preoedipal transference, countertransference, and the body. PMID- 8226180 TI - Collaboration and conflict: Ernest E. Hadley and Harry Stack Sullivan, 1930-1945. PMID- 8226181 TI - Sex in a Confucian society. PMID- 8226182 TI - A young woman's homosexuality reconsidered: Freud's "The psychogenesis of a case of homosexuality in a woman". PMID- 8226183 TI - Some transatlantic reflections on language in psychoanalysis. PMID- 8226184 TI - The use of psychodynamic psychotherapy in manic-depressive illness. PMID- 8226185 TI - Vagina envy in men. PMID- 8226186 TI - [Treklovern--Family-Maternity facility in Uppsala]. PMID- 8226187 TI - [Prevention of excess neonatal morbidity associated with Group B streptococci by vaginal chlorhexidine disinfection during labor]. PMID- 8226188 TI - [Swedish Obstetrics and Gynecology Association and Swedish Midwifery Society recommend physician-midwife team work in delivery rooms]. PMID- 8226189 TI - [Acupuncture during pregnancy and delivery--a midwife's concern]. PMID- 8226190 TI - [Are both nurses and midwives to be found in preventive activities?]. PMID- 8226191 TI - [Are midwives their own worst enemies?]. PMID- 8226192 TI - [Why do women go around the maternity ward dressed in bathrobe and with an identification band around the arm--and why do infants lie in transparent plastic beds with wheels?]. PMID- 8226193 TI - [Active physical training during pregnancy]. PMID- 8226194 TI - [Childbirth preparation in water]. PMID- 8226195 TI - [Female university students in Sweden: sex, contraceptives and STD]. PMID- 8226196 TI - [Left-sided delivery position according to Simm's method]. PMID- 8226197 TI - [Comments concerning SBF's and SFOG's consensus report on physician-midwife team work in delivery rooms]. PMID- 8226198 TI - [Acupuncture for analgesia during childbirth]. PMID- 8226199 TI - International Code of Ethics for Midwives. PMID- 8226201 TI - [Reading is necessary!]. PMID- 8226200 TI - [Report of the ICM Council meeting, 4-6 May 1993, Vancouver Canada]. PMID- 8226202 TI - [Midwives' youngster. Address at the 100th anniversary of Halsingland's Midwifery Association on 7 November 1993 in Hotel Jarvsobaden in Jarvso]. PMID- 8226203 TI - [Maternal-Infant care in a refugee camp. Report from a refugee camp in southern Lebanon]. PMID- 8226204 TI - [Welfare and quality of care in Taiwan and China]. PMID- 8226205 TI - [Report from a conference in Taiwan]. PMID- 8226206 TI - Farm animal welfare: progress on several fronts. PMID- 8226207 TI - Concerned about unlicensed practice. PMID- 8226208 TI - Pain control. PMID- 8226209 TI - Thoughts provoked by symposium on extra-label drug use. PMID- 8226210 TI - Some implications of structural change in veterinary medicine and its impact on veterinary education. PMID- 8226211 TI - A soldier's story: my recollection of veterinary life in WW II. PMID- 8226212 TI - What is your diagnosis? Fat opacity within the soft tissues. Bony involvement was not evident. PMID- 8226213 TI - ECG of the month. Incomplete interference AV dissociation with a junctional rhythm in a horse. PMID- 8226214 TI - Animal behavior case of the month. PMID- 8226215 TI - An insurer's liability for judgments in excess of policy limits. PMID- 8226216 TI - Evaluation of a one-step surgical preparation technique in dogs. AB - The efficacy of a 1-step surgical preparation technique for skin of dogs prior to elective ovariohysterectomy was evaluated. Dogs randomly assigned to group 1 (n = 30) had their skin prepared for surgery by use of a 2-step method, whereas the skin of dogs in group 2 (n = 30) was prepared for surgery by use of a commercially available product for a 1-step technique. Culture plates for quantitative bacterial counts were applied to the proposed incision site on dogs under general anesthesia after hair at the site was clipped and vacuumed but before antiseptic was applied. A second quantitative bacterial culture plate was applied to the proposed incision site after completion of the surgical preparation technique. Surgeries were routinely completed, and dogs were evaluated by physical examination the next day and at the time of suture removal (7 to 10 days after surgery) for complications. Postoperative complications were minor and consisted primarily of subcutaneous swelling, which resolved with time. All cultures obtained prior to skin preparation included bacteria or yeast. Sixteen cultures obtained after skin preparation (group 1, n = 11; group 2, n = 5) included bacteria or yeast. The total number of colonies of potential pathogens (Staphylococcus sp and Enterobacteriaceae) on the preparation cultures was 9,339; 4 colonies were counted on the postpreparation cultures. Potential bacterial pathogens, ie Streptococcus intermedius and gram-negative bacteria, were isolated from dogs prepared with the 2-step technique.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226217 TI - Left aortic arch and right ligamentum arteriosum causing esophageal obstruction in a dog. AB - A 10-week-old, mixed-breed dog was examined because of persistent regurgitation. Results of thoracic radiography and fluoroscopy of the esophagus, using a liquid suspension of barium sulfate, were suggestive of a vascular ring anomaly. During left lateral thoracotomy, a left aortic arch was identified, but the cause of the esophageal obstruction could not be determined. Results of endoscopic examination of the esophagus suggested a compressive lesion originating from the right side of the esophagus. An anomalous artery could not be identified on an aortogram. Post-mortem examination revealed a left aortic arch and right ligamentum arteriosum that were causing the esophageal obstruction. PMID- 8226218 TI - Complications after ultrasound-guided biopsy of abdominal structures in dogs and cats: 246 cases (1984-1991). AB - Medical records of 195 dogs and 51 cats that underwent a total of 233 ultrasound guided biopsies and 70 ultrasound-guided fine-needle aspirations were reviewed for diagnostic quality of the specimens obtained and for procedural complications. Three animals (1.2%) had major postbiopsy complications. All 233 animals that underwent ultrasound-guided biopsy were examined ultrasonographically immediately after biopsy. Thirteen (5.6%) were determined to have minor localized hemorrhage. Of the 233 biopsy specimens, 226 (97%) were considered to be of adequate quality for histologic interpretation. Results of cytologic examination of 59 (84.3%) of 70 specimens obtained by fine-needle aspiration correlated with the final diagnosis made during surgical exploration or at necropsy. PMID- 8226219 TI - Transoral axial division, under endoscopic guidance, for correction of epiglottic entrapment in horses. AB - Transoral axial division was performed, using endoscopic guidance, in 20 horses with epiglottic entrapment. Mean age of affected horses was 3 years (range, 2 to 5 years), and there were 18 Standardbreds and 2 Thoroughbreds. Standardbreds and female horses were significantly overrepresented when compared with a reference population. Upper respiratory noise prompted endoscopic examination before surgery in 16 horses, but noise was not heard in 4 horses. Ten horses had entrapment by thin tissue without ulceration, 6 horses had entrapment by thin tissue with small, partial-thickness ulceration, 3 horses had entrapment by moderately thick tissue with large, partial-thickness ulceration, and 1 horse had entrapment with thick tissue and severe ulceration. After surgery, horses received anti-inflammatory medication and box stall rest with hand walking for 7 days. All horses returned to race training on day 7 after surgery and returned to their previous level of racing or training. Epiglottic entrapment recurred in 2 horses (10%), and dorsal displacement of the palate developed in 2 horses after surgery. Relationship of epiglottic entrapment and dorsal displacement of the palate to epiglottic hypoplasia was not apparent. Transoral axial division under endoscopic guidance appeared to be a useful, safe, and economic alternative to currently available methods for management of epiglottic entrapment in horses. Horses with entrapment by thick and ulcerated tissue may be best managed by surgical resection of entrapping tissue. PMID- 8226220 TI - Possible normokalemic variant of hyperkalemic periodic paralysis in two horses. AB - Hyperkalemic periodic paralysis (HPP), characterized by intermittent episodes of muscle fasciculations, profound muscle weakness, and hyperkalemia, has been described in Quarter Horses, Appaloosas, and Paints. In previous reports, the hallmark of this syndrome has been the development of hyperkalemia during each episode. Two affected horses had episodes of paralysis without associated hyperkalemia, demonstrating that normokalemia during an episode otherwise consistent with HPP does not eliminate HPP as a diagnosis. This clinical presentation appeared to be a variant of HPP. PMID- 8226221 TI - Left dorsal displacement of the colon with splenic adhesions in three horses. AB - Three horses underwent exploratory celiotomy because of signs of acute abdominal pain. At surgery, all horses were diagnosed as having left dorsal displacement of the large colon. Each surgery was complicated by fibrous adhesions of the spleen to the body wall. All horses had previously undergone abdominal surgery for colic. In these cases, it appeared that the left large colon displaced cranially and then entered the renosplenic space from a cranial to caudal direction. Adhesions of the spleen to the body wall would prevent correction of left dorsal displacement of the large colon by the rolling technique. PMID- 8226222 TI - Extensor tendon lacerations in horses: 50 cases (1982-1988). AB - Medical records of 50 horses with extensor tendon lacerations affecting 53 limbs over 7 years (1982 to 1988) were reviewed to determine the clinical features and prognosis for athletic soundness after treatment. Mean and median ages were 4.8 and 3 years, respectively. Eighty-nine percent of the injuries involved the hind limbs, with approximately equal distribution between the right and left limbs. Hind limb lacerations involved the long digital extensor tendon (28 limbs), lateral digital extensor tendon (3 limbs), or both tendons (16 limbs). Forelimb lacerations involved the common digital extensor tendon (2 limbs), lateral digital extensor tendon (1 limb), or both tendons (3 limbs). Joint involvement was found in 6 cases. Wound management varied according to wound characteristics, financial constraints of owners, and clinician preference. External coaptation consisted of a 3-layered cotton bandage in 33 limbs, splint and cotton bandage in 12 limbs, fiberglass or plaster-of-Paris cast in 5 limbs, and no form of coaptation in 3 limbs. Follow-up information for 40 cases at least 1 year after injury revealed that 29 (73%) horses had returned to athletic soundness, 7 (18%) horses were pasture sound, 3 horses were chronically lame, and 1 horse was euthanatized during treatment because of wound sepsis. Depending on the hind limb tendons involved, return to athletic soundness ranged from 62% (long digital and lateral digital extensor tendons lacerated) to 80% (long digital extensor tendon lacerated) The small number of forelimb lacerations precluded any comparative data.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226223 TI - Fetal loss associated with palpation per rectum to diagnose pregnancy in cows. AB - A prospective observational study was performed to determine whether palpation per rectum of cows in the first 6 weeks of gestation to diagnose pregnancy contributed to fetal attrition. Pregnancy diagnoses were made by private practitioners as part of their routine herd-health service on 9 dairies in the San Joaquin Valley of California. To determine whether there was an association between abortion and fetal age at time of palpation, the probability of abortion was tested as a function of fetal age at palpation, controlling for possible modifying and confounding effects of herd, age at conception, gravidity, parity, and number of days-in-lactation at conception. Results of logistic regression analyses for 19,411 pregnancies followed for up to 90 days after palpation indicated that, during the 28- to 42-day period, palpation of fetuses earlier in the period was associated with a significantly (P < 0.0001) low probability of abortion, compared with that for palpation later in the period. An association between abortion and palpation of fetuses > 42 days of age was not found. Results were suggestive that, given conditions and techniques typical of private practice, fetal death may not be a usual manifestation of early palpation of cows to diagnose pregnancy, rather, that there may be a slight increase in risk of fetal death as the fetal age at palpation increases from 28 to 42 days. PMID- 8226224 TI - Successful treatment of retained placenta with umbilical cord injections of collagenase in cows. AB - Fetal membranes usually are released from the uterus between 2 and 6 postpartum hours. However, in a substantial percentage of cows (11%), fetal membranes are retained for several days. In part, failure of collagen breakdown seems to be related to retention of fetal membranes. Injections of 200,000 U of bacterial collagenase in 1,000 ml of physiologic saline solution via umbilical arteries (1 or 2) between 24 and 72 hours of retention caused release of retained fetal membranes in 23 of 27 cows (85%) with noninduced retained fetal membranes and in 10 of 14 cows (71%) with experimentally induced retained fetal membranes, within 36 hours after injection. Controls (n = 36) did not release retained fetal membranes within this time. Injections of collagenase via a jugular vein (2.2 x 10(6) U in 1,000 ml of physiologic saline solution), administered over a 30 minute period, caused release of retained fetal membranes within 36 hours in 3 of 6 cows with experimentally induced retained fetal membranes. Clinical complications did not follow treatments with collagenase. Umbilical injections of bacterial collagenase were highly effective in the treatment of retained fetal membranes in cows. The procedure is simple, safe, affordable, and can be completed in 25 minutes. PMID- 8226225 TI - Neutering of feral cats as an alternative to eradication programs. PMID- 8226226 TI - More on early-age neutering. PMID- 8226227 TI - Animal behavior. PMID- 8226228 TI - Article on Borrelia burgdorferi revisited. PMID- 8226229 TI - Hiring the recent graduate. PMID- 8226230 TI - Endoscopic placement of a percutaneous gastroduodenostomy feeding tube in dogs. PMID- 8226231 TI - What is your diagnosis? Mineralized mass of the axis and mineralized masses of the right first rib, right second rib, and right eighth rib. PMID- 8226232 TI - Bilateral stress fractures of the tibia in a racing American quarter horse. PMID- 8226233 TI - Selected employment characteristics in public or corporate sectors of the veterinary medical profession. PMID- 8226234 TI - Enterprise analysis: a method to evaluate alternative production and management practices for cow/calf operations. AB - Food animal veterinarians recognize the need to economically justify their recommendations regarding whole-herd management programs. An enterprise analysis technique may be used by veterinarians to determine the actual cost of production on beef cattle operations. Enterprise analysis was used on 2 groups of cattle to demonstrate the financial impact of a recommendation to modify the winter supplementation program of a large ranch. Pregnancy rates improved from 62 to 95% for group-1 cattle and from 75 to 94% for 2 groups of cattle in the 2-year study. Additionally, the calving cycle was shortened and the number of calves born earlier in the calving season was notably increased, which raised the total pounds of calves sold at weaning. These outcomes were realized without additional winter-supplementation expenses for either group. Veterinarians can use enterprise analysis to determine the value of their recommendations, thus allowing them to charge more appropriately for the services provided. PMID- 8226235 TI - Effect of an orally administered sulfonylurea, glipizide, for treatment of diabetes mellitus in cats. AB - An orally administered sulfonylurea drug, glipizide, was evaluated for treatment of diabetes mellitus. Confirmation of diabetes was based on evidence of appropriate clinical signs, persistent hyperglycemia, and glucosuria. Glipizide (5 mg, PO, q 12 h) was administered to each cat. Sixteen cats were fed a commercial high-fiber diet and 4 cats were fed a commercial low-fiber diet. Insulin was not administered to any cat during the study. Each cat was evaluated 2, 4, 8, and 12 weeks after initiation of treatment. Three clinical responses to glipizide treatment were identified. Mean preprandial blood glucose concentration and mean blood glucose concentration during an 8-hour postprandial period decreased to < 200 mg/dl in 5 of 20 (25%) cats. In these 5 cats, glucosuria was no longer detected and clinical signs resolved by the 4-week reevaluation. Euglycemia was maintained after discontinuing glipizide treatment in 2 of these 5 cats. Glycemic control has been maintained in 2 of 5 of the responding cats for 5 and 7 months of glipizide treatment. One of 5 of the responding cats developed insulin-requiring diabetes mellitus after 6 months of glipizide treatment. Seven of 20 (35%) cats failed to respond to treatment. Mean preprandial blood glucose concentration and mean blood glucose concentration during an 8-hour postprandial period did not change from pretreatment values after 2 +/- 1 months; glucosuria persisted and clinical signs progressively worsened. Insulin treatment was required to establish glycemic control in these 7 cats. Eight of 20 (40%) cats partially responded to glipizide treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226236 TI - Onchocerciasis in two dogs. AB - Although widely reported in cattle, horses, and other ungulates, onchocerciasis has only recently been reported in 1 dog. We report 2 additional dogs with onchocerciasis involving the palpebral conjunctiva, third palpebra, and sclera. Both dogs were evaluated because of ocular or periocular masses. Histologically, viable adult parasites were surrounded by minimal fibrosis and few macrophages. Pyogranulomas were often centered around degenerating or mineralized Onchocerca organisms. Onchocerciasis should be considered in the differential diagnosis of ocular or periocular nodules in dogs, particularly dogs from western states. PMID- 8226237 TI - Sudden death associated with Neospora caninum myocarditis in a dog. AB - A 10-month-old female Bull Mastiff dog was found dead without reports of any prior clinical signs of disease. Necropsy revealed lesions in the myocardium, which consisted of a large 2 x 2.3-cm brownish area and a few small similar lesions. Microscopically, Neospora caninum tachyzoites were found in the lesions characterized by necrosis and interstitial myocarditis. Diagnosis was confirmed by immunohistochemical staining with anti-N caninum serum. PMID- 8226238 TI - Clinical and laboratory findings in chronic conjunctivitis in cats: 91 cases (1983-1991). AB - Clinical findings and laboratory test results from 91 cats with chronic conjunctivitis were studied to determine the causes of the disease and the sensitivity of diagnostic procedures used, and to identify correlations between results of various diagnostic procedures and clinical or signalment variations. Mean age of affected cats was 2.9 +/- 2.7 years (+/- SD), with a range from 1 month to 11 years. Conjunctivitis was more likely to be bilateral (56 cats) than unilateral (35 cats). In cats tested for FeLV or feline immunodeficiency virus infection, 15 and 8.5%, respectively, of the results were positive, compared with 4 and 2.6% for the general hospital population. Culturing or immunofluorescent assay (IFA) for feline herpesvirus 1 (FHV-1) and Chlamydia psittaci IFA resulted in identification of pathogens (positive test results) in 19% (FHV-1) and 18% (C psittaci) of tested cats. For FHV-1, culturing was more sensitive than was IFA, yielding positive results in 19 vs 8.8% of cases. In only 1 cat were FHV-1 and chlamydiae recovered. The probability of positive test results for FHV-1 or chlamydiae was unrelated to concurrent corneal disease, unilateral vs bilateral involvement, or age. Cause of conjunctivitis could not be definitively determined in the remaining 35 cases tested for both agents. Bacterial species considered to be potentially pathogenic were isolated from conjunctival sac specimens in only 1 of 38 attempts. Cytologic changes considered compatible with chlamydial or FHV-1 infection (intracytoplasmic inclusions or multinucleated epithelial cells, respectively) were found in 8 and 5 cases, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226239 TI - Ultrasonographic abnormalities in the eyes of dogs with cataracts: 147 cases (1986-1992). AB - Records of 147 dogs with cataracts and that had an ultrasonographic examination of their eyes were reviewed. Ultrasonography was performed, using a real-time mechanical sector scanner (7.5 or 10.0 MHz transducer), after topical anesthesia of the cornea. Vitreous degeneration was diagnosed in 23% of the eyes examined. Retinal detachment was detected in 11% of all eyes, but was detected in 4% of eyes with immature cataracts, 6.5% of eyes with mature cataracts, and 19% of eyes with hypermature cataracts. In 66% of the eyes, the cataract was the only ultrasonographic abnormality found. Vitreous degeneration and retinal detachment were most often observed in eyes with a hypermature cataract, and were uncommon in eyes with an immature cataract. We concluded that ultrasonographic examination can detect abnormalities of the posterior segment when opacity of the anterior segment precludes complete ophthalmoscopic examination and that it is a quick and easy procedure for screening dogs for retinal detachment prior to cataract surgery. PMID- 8226240 TI - Hypocalcemia at parturition as a risk factor for left displacement of the abomasum in dairy cows. AB - Total and ionized serum calcium concentrations were measured in 510 Holstein cows at the time of parturition. Cows that subsequently developed left displacement of the abomasum (LDA) were compared with those that did not. Cows that were hypocalcemic at parturition had a 4.8 times greater risk of developing LDA than did normocalcemic cows. Hypocalcemia was associated with the development of LDA whether ionized serum calcium concentration or total serum calcium concentration was used to determine whether cows were hypo- or normocalcemic. The proportion of cows developing LDA was not significantly different between cows that had clinical signs of hypocalcemia and were treated for parturient paresis and cows that were hypocalcemic, but did not have clinical signs and were not treated. PMID- 8226241 TI - Gentamicin toxicosis in a North American cougar. AB - This report documents nephrotoxicosis in a North American cougar (Felis concolor) caused by gentamicin overdose and describes a method of calculating drug dosages by metabolic scaling for species in which pharmacokinetic studies have not been performed. Determining drug dosages by metabolic scaling can decrease the likelihood of administering inappropriate drug doses in exotic animals. PMID- 8226242 TI - Metabolic and genetic aspects of urate urolithiasis in Dalmatians. PMID- 8226243 TI - Diagnosis, prevention, and treatment of urate urolithiasis in Dalmatians. PMID- 8226244 TI - Veterinarians as animal advocates during disasters: the California Veterinary Medical Association's disaster response program. PMID- 8226245 TI - Veterinary service and animal care emergency operations plans. PMID- 8226246 TI - AVMA emergency preparedness planning. PMID- 8226247 TI - Epidemiology of cranial cruciate ligament rupture in dogs. AB - Data from 10,769 dogs with rupture of the cranial cruciate ligament (CCL) were compared with data from a control population of 591,548 dogs to determine whether age, breed, gender, or body weight was associated with prevalence of CCL rupture. Prevalence of CCL rupture increased as dogs became older, with peak prevalence in dogs 7 to 10 years old. Among breeds represented by > 1,000 individuals, Rottweilers, Newfoundlands, and Staffordshire Terriers had the highest prevalence of CCL rupture, whereas Dachshunds, Basset Hounds, and Old English Sheepdogs had the lowest. Neutered dogs, whether male or female, had a higher prevalence of CCL rupture than did sexually intact dogs. The dog's age at the time of ovariohysterectomy was not associated with prevalence of CCL rupture. Dogs weighing > 22 kg had a higher prevalence of CCL rupture, compared with dogs weighing < 22 kg, and tended to rupture their CCL at a younger age. PMID- 8226248 TI - Hypertrophic cardiomyopathy in a dog. AB - A 12-year-old spayed Corgi with heart failure did not respond to conventional medical therapy for congestive cardiomyopathy (furosemide, 2.5 mg/kg of body weight, IV, q 8 h; captopril 1 mg/kg, PO, q 8 h). Cardiac ultrasonography subsequently revealed a hypertrophic left ventricle and interventricular septum. Because a causative agent could not be identified, a diagnosis of primary hypertrophic cardiomyopathy was made. Administration of diltiazem (1.25 mg/kg, PO, q 12 h) resulted in sustained clinical improvement for several weeks. Complete cardiac evaluation, including echocardiography, was essential in determining the correct treatment for this dog. PMID- 8226249 TI - Effects of diazepam, acepromazine, detomidine, and xylazine on thiamylal anesthesia in horses. AB - The cardiorespiratory effects of thiamylal (10 mg/kg of body weight, IV) and the effects of preanesthetic medication with diazepam, acepromazine, detomidine, or xylazine administered prior to a thiamylal dosage of 6 mg/kg, IV, were evaluated in 6 adult horses. The quality of recovery from thiamylal anesthesia also was evaluated. Intravenous administration of thiamylal at a dosage of 10 mg/kg increased heart rate, systemic arterial, pulmonary artery, and central venous blood pressures, as well as cardiac output and arterial partial pressure of CO2 (PaCO2). The maximal rate of right ventricular pressure increase (RVdP/dtmax), respiratory rate, and arterial partial pressure of O2 (PaO2) decreased, whereas arterial pH and systemic vascular resistance remained unchanged. Preanesthetic medication with diazepam prior to IV administration of thiamylal (6 mg/kg) did not change the pattern of this response, but diazepam did increase heart rate, cardiac output, and respiratory rate during the recovery period. Administration of acepromazine (0.1 mg/kg, IV) prior to administration of thiamylal increased heart rate and decreased systemic arterial and central venous blood pressures and systemic vascular resistance. Detomidine (10 micrograms/kg, IV), administered prior to thiamylal, decreased heart rate, cardiac output, and respiratory rate, and increased right atrial blood pressure. Administration of xylazine (0.5 and 1.0 mg/kg, IV) prior to thiamylal induced effects qualitatively similar to detomidine. Thiamylal decreased RVdP/dtmax and PaO2 in horses that received diazepam, acepromazine, detomidine, or xylazine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226250 TI - Granulomatous enteritis and Campylobacter bacteremia in a horse. AB - A 10-month-old Standard-bred colt was examined because of weight loss, fever, and extensive subcutaneous edema of the ventral aspects of its trunk. Diagnosis of granulomatous enteritis was based on D-xylose malabsorption, hypoproteinemia, and rectal mucosal biopsy. Campylobacter fetus subsp fetus was cultured from jugular venous blood when the horse was admitted to the hospital and 24 hours later. Appropriate antimicrobial agents and glucocorticoids failed to effect clinical remission. After euthanasia, granulomatous enteritis was confirmed at necropsy but C fetus subsp fetus was not isolated from tissues. PMID- 8226251 TI - Effects of bilateral ovariectomy via colpotomy in mares: 23 cases (1984-1990). AB - Postoperative performance and behavioral patterns were investigated retrospectively in 23 client-owned mares after bilateral ovariectomy via colpotomy. The interval from surgery to postoperative inquiries ranged from 9 to 67 months. Information obtained from review of the medical record and client interviews included the reason for ovariectomy, postoperative complications, problems identified by owners after discharge of the mare from the hospital, postoperative level of athletic performance, postoperative signs of estrus, and overall owner satisfaction. Reasons given by owners for having mares ovariectomized were behavioral modification (16 mares), use as embryo-transfer recipients (3 mares), use as mount mares for collecting semen (2 mares), elimination of chronic colic during estrus (1 mare), and sterilization for registration (1 mare). Postoperative complications developed in 4% (1/23) of the mares; however, problems were noticed by the owners of 4 other mares after discharge from the hospital. Continuing signs of behavioral estrus were detected in 35% (8/23) of the mares, but in only in 9% (2/23) was the behavior judged to be objectionable by the owner. Of 12 mares used in performance events prior to bilateral ovariectomy, 10 were judged to be competing at greater than preoperative levels, 1 was judged to be competing at the same level, and 1 was judged to be competing at less than preoperative level of performance. Of 18 owners, 14 were satisfied, 2 were undecided, and 2 were dissatisfied with their mare after it had had bilateral ovariectomy. PMID- 8226252 TI - Veterinary colleges explore diversity. PMID- 8226254 TI - Animal welfare, import/export: issues in Australia. PMID- 8226253 TI - Quality assurance programs receive thumbs up from insurance trust. PMID- 8226255 TI - Behavioral services in a practice lead to quality relationships. PMID- 8226256 TI - Questions designation of feline practice as a new specialty. PMID- 8226257 TI - Support optimal animal care everywhere. PMID- 8226258 TI - Steel-jaw leghold trap springs again. PMID- 8226259 TI - Potential roles of veterinary schools in emergency and disaster management. PMID- 8226260 TI - Diversity: a veterinary student's outlook. PMID- 8226261 TI - Fulfilling the dream for foreign adventure. PMID- 8226262 TI - What is your diagnosis? Well-delineated, uniformly mineralized mass, without evidence of invasion of or origination from any of the regional bony structures. PMID- 8226263 TI - ECG of the month. PMID- 8226264 TI - Animal behavior case of the month. PMID- 8226266 TI - Symposium on disaster medicine. Proceedings. PMID- 8226265 TI - Using small claims affidavits. PMID- 8226267 TI - Lessons learned from Hurricane Andrew. PMID- 8226268 TI - Hurricane Andrew: the importance of coordinated response. PMID- 8226269 TI - Business aspects of disaster planning and recovery. PMID- 8226270 TI - The Maryland pet-sheltering plan. PMID- 8226271 TI - The Maryland plan for disaster recovery: disposal of dead animals. PMID- 8226272 TI - Use of search and rescue dogs. PMID- 8226273 TI - A very low dose of green tea polyphenols in drinking water prevents N-methyl-N nitrosourea-induced colon carcinogenesis in F344 rats. AB - The effect of tea polyphenols, major constituents of tea, on colon carcinogenesis was investigated. A total of 129 female F344 rats were given an intrarectal instillation of 2 mg of N-methyl-N-nitrosourea 3 times a week for 2 weeks, and received a water solution of green tea extract (GTE) as drinking water throughout the experiment. Autopsies at week 35 revealed significantly lower incidence of colon carcinomas in rats ingesting 0.05%, 0.01% or 0.002% GTE solution than in controls ingesting 0% GTE solution: 43%, 40% and 33% vs. 67%. The data suggest that GTE, even at a very low dose (0.002% solution), has a potent inhibitory effect on colon carcinogenesis. PMID- 8226274 TI - Influence of esculetin on incidence, proliferation, and cell kinetics of mammary carcinomas induced by 7,12-dimethylbenz[a]anthracene in rats on high- and low-fat diets. AB - The effects of a high-fat diet and esculetin were investigated on 7,12 dimethylbenz[a]anthracene (DMBA)-induced mammary carcinogenesis in female Sprague Dawley rats. Rats were given a 5-mg dose of DMBA. Seven days later, they were fed either a high-fat (20% soybean oil) or low-fat (0.5% soybean oil) diet. A half of the rats received diets containing 0.03% esculetin. Esculetin significantly inhibited tumor incidence, growth and cell kinetics of the tumor in the rats fed the high-fat and the low-fat diets. Our findings indicate that DMBA-induced mammary tumorigenesis is affected by lipoxygenase products. PMID- 8226275 TI - Primary gastric carcinoma cells frequently lose heterozygosity at the APC and MCC genetic loci. AB - Loss of heterozygosity (LOH) at APC and MCC gene loci (both mapped to 5q21) was investigated in 24 surgical specimens of primary gastric carcinomas using the polymerase chain reaction after tumor cell enrichment by cell sorting based on differences in DNA content. LOH at APC and/or MCC was detected in 87% (13/15) of the cases; at the APC in 86% (12/14) and at the MCC locus in 100% (7/7). LOH at the APC locus was always accompanied by LOH at the MCC locus. LOH at the APC and/or MCC was found in both differentiated and undifferentiated types in both early and advanced stages of gastric carcinoma. Thus, LOH at APC and/or MCC is considered to be one of the most prevalent genetic alterations in human gastric carcinoma and occurs at an early stage of the carcinogenesis. PMID- 8226276 TI - Absence of linkage between the retinoblastoma gene and hts gene in the LEC rat: a model of human Wilson's disease. AB - The LEC rat is an authentic model of human Wilson's disease (WD) with an autosomal recessively inherited hepatitis. We investigated linkage between the hepatitis gene (hts) and the rat retinoblastoma gene (RB), that is closely linked to the WD gene in humans, to see whether or not the hts gene is located on the syntenic region of WD and is the counterpart of the WD gene. Polymerase chain reaction-single strand conformation polymorphism analysis with backcross progenies from LEC and TM strains showed that the recombination rate between these two loci was 55.6%, demonstrating that the hts and RB genes are not linked to each other. These data indicate that the hts gene is not the counterpart of the WD gene and that the human syntenic region on which the WD locus and human RB gene are located, is not conserved in the rat genome. PMID- 8226277 TI - Synergistic effect of radiation on colon carcinogenesis induced by methylazoxymethanol acetate in ACI/N rats. AB - The effect on colon and liver carcinogenicity in rats of a single X-irradiation exposure given either before or after methylazoxymethanol (MAM) acetate was studied in ACI/N rats of both sexes. A single dose of X-irradiation (3 Gy) was administered either 3 months before or after three weekly s.c. injections of MAM acetate (25 mg/kg body weight). At 365 days after the start, the incidence and multiplicity of MAM acetate-induced intestinal tumors were enhanced by X irradiation either prior to or after the MAM acetate treatment. In addition, X irradiation before MAM acetate increased the incidence of hepatocellular foci in either sex. In females, X-irradiation either before or after MAM acetate exposure decreased intestinal tumorigenesis. These findings suggest an apparent synergism of these agents in intestinal carcinogenesis of male rats. PMID- 8226278 TI - Influence of gastric pH modifiers on development of intestinal metaplasia induced by X-irradiation in rats. AB - The influence of gastric pH on intestinal metaplasia was examined in male Crj:CD(SD) rats. At the age of 5 weeks, animals were irradiated with two 10 Gy doses of X-rays to the gastric region at a 3-day interval (total 20 Gy), and 6 months after irradiation, received either secretin or histamine in silicon tubes for 2 months or had their bilateral submandibular salivary glands removed. The incidence of intestinal metaplasia in the fundus of animals after administration of secretin or histamine, or removal of the salivary glands were reduced, along with the pH values, as compared with values for rats given X-rays alone. In both the pyloric and the fundic gland mucosae, the numbers of alkaline phosphatase (ALP)-positive foci and type B metaplasias (intestinal crypts without Paneth cells) were also significantly decreased (P < 0.01). In a second experiment, started six months after irradiation, rats were kept on 1% sodium chloride (NaCl) diet for 6 months. Subsequent removal of salivary glands along with histamine treatment brought about a marked drop in pH and in numbers of ALP-positive foci after three and five days. The present results thus indicated that development and maintenance of intestinal metaplasia can be influenced by a decrease of pH value. PMID- 8226279 TI - Growth dependence of human papillomavirus 16 DNA-positive cervical cancer cell lines and human papillomavirus 16-transformed human and rat cells on the viral oncoproteins. AB - The dependence on human papillomavirus (HPV) oncoproteins of the growth of cervical cancer cell lines [C4-1, HeLa (both containing HPV 18 DNA), CaSki and SiHa (both containing HPV 16 DNA)], HPV 16-transformed human embryonic kidney cells, and HPV 16-transformed rat brain and 3Y1 cells was examined by using antisense RNA approaches. The cells were transfected with plasmids expressing RNA antisense to the HPV 16 or 18 open reading frames E6E7, together with plasmids expressing the hygromycin B resistance gene, and drug-resistant colonies were scored three weeks later. In all the human cell lines, the efficiency of colony formation was lowered by RNA antisense to the resident HPV type. Some of the rat cell lines responded to the antisense plasmids, but some did not. From a nonresponding rat tumor line (3Y1HP-1T), cell clones with various levels of E7 protein were isolated after transfection with the antisense plasmid, and were examined for anchorage-independent growth in soft agar. The colonies formed by the clones with lower E7 levels tended to be smaller and fewer than those formed by the clones with higher E7 levels. These findings strongly suggest that some of the transformed or cancer phenotypes of cells in vitro are dependent, even after extensive passages and malignant changes, on expression of the oncoproteins of the resident HPV. PMID- 8226280 TI - Establishment and characterization of BALB/c retroperitoneal sarcoma with spontaneous liver metastases. AB - The objective of this study was to examine the identity and characteristics of a spontaneously occurring murine retroperitoneal tumor of BALB/c mouse origin that selectively metastasized to the liver. From the primary tumor, a permanent cell line, termed LMFS (liver metastasis from sarcoma) was established in vivo and in vitro. After a subcutaneous injection of more than 1 x 10(5) cells in the side back of mice, the LMFS cells proliferated at the inoculation site (100% take) and induced metastatic nodules spontaneously in the liver, but not in the lung. By the limiting dilution technique, a cloned cell line, LMFS-1, was established in vitro. The LMFS-1 cell line had similar morphological characteristics to the LMFS cells both in vitro and in vivo. The doubling time of the LMFS-1 cell line was 10 h in passage 60. The number of chromosomes ranged from 71 to 108 and 93% of metaphases showed near-tetraploidy. In microscopic examination, no specific arrangement of the LMFS tumor cells was seen; the LMFS cell had medium- to large sized atypical nuclei and clear and large cytoplasm. Electronmicroscopy showed that the cytoplasm of the LMFS cell had a moderate amount of rough-surfaced endoplasmic reticulum but no desmosomes or microvilli. Immunohistochemically, the LMFS cells were positive for vimentin, but showed no reaction for keratin or cytokeratin. Therefore, the LMFS tumor was considered to be an undifferentiated sarcoma. The LMFS cell line should be a useful tool not only for studies of metastasis, but also for experiments on the therapy of hepatic tumors. PMID- 8226281 TI - Triflavin, an Arg-Gly-Asp-containing peptide, inhibits tumor cell-induced platelet aggregation. AB - In this study, we examined the effect of triflavin, an Arg-Gly-Asp (RGD) containing snake venom peptide, on human cervical carcinoma (HeLa) cell- and B16 F10 mouse melanoma cell-induced platelet aggregation (TCIPA) in heparinized platelet-rich plasma. TCIPA appears to play an important role in the development of certain experimental tumor metastases. Two ADP-scavenging agents, apyrase (10 U/ml) and creatine phosphate (CP) (5 mM)/creatine phosphokinase (CPK) (5 U/ml) completely inhibited B16-F10 TCIPA, but hirudin (5 U/ml) had no effect. In contrast, apyrase and CP/CPK did not inhibit HeLa TCIPA while hirudin completely inhibited it. Furthermore, HeLa cells initially induced platelet aggregation and then blood coagulation at a later stage. In addition, HeLa cells shortened, in a concentration-dependent manner, the recalcification time of normal as well as factor VIII- and IX-deficient human plasma, but did not affect the recalcification time of factor VII-deficient plasma. This suggests that HeLa TCIPA occurs via activation of the extrinsic pathway, probably owing to tumor cell expression of tissue factor-like activity. HeLa cell-induced thrombin generation was confirmed by detection of amidolytic activity towards a chromogenic substrate, S-2238 (H-D-Phe-Pip-Arg-p-NA). Triflavin and GRGDS inhibited, in a dose-dependent manner, TCIPA caused by either cell line. On a molar basis, triflavin was 10,000-30,000 times more potent than GRGDS in this regard. Moreover, monoclonal antibodies raised against glycoprotein (GP) IIb/IIIa complex (i.e., 7E3 and AP2) and against GP Ib (i.e., AP1) completely inhibited HeLa TCIPA. 7E3 and AP2 inhibited B16-F10 TCIPA by up to 80% whereas AP1 showed only 30% inhibition of B16-F10 TCIPA. In conclusion, the inhibitory effect of triflavin on HeLa and B16-F10 TCIPA may be mediated principally by the binding of triflavin to the fibrinogen receptor associated with GP IIb/IIIa complex on the platelet surface. However, GP Ib is also involved in HeLa TCIPA as thrombin formation is the key factor in triggering platelet aggregation caused by HeLa cells. PMID- 8226282 TI - Enhanced antitumor activity and reduced toxicity of 1,3-bis(2-chloroethyl)-1 nitrosourea administered in lipid microspheres to tumor-bearing mice. AB - Stable lipid microspheres (LM) and lipid nanospheres (LN) with average diameters of 200 nm and 50 nm, respectively, were used to encapsulate an lipophilic antitumor agent, 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). LM and LN containing BCNU (lipo BCNU and s-lipo BCNU, respectively) were prepared by homogenizing a soybean oil solution of BCNU with egg yolk lecithin, and their antitumor activity via the intravenous route was tested against L1210 leukemia in mice and compared with that of BCNU dissolved in saline. Both lipo-BCNU and s lipo BCNU showed significantly enhanced antitumor activity with reduced toxicity, when compared with the corresponding doses of BCNU alone. These results suggest that LM and LN may be suitable carriers for lipophilic antitumor agents and may enhance their efficacy. PMID- 8226283 TI - Clinical application of serum pepsinogen I and II levels for mass screening to detect gastric cancer. AB - A considerable number of gastric cancers derive from stomach mucosa where chronic atrophic gastritis is severe and extensive. Based on the fact that the serum pepsinogen levels provide a precise measure of the extent of chronic atrophic gastritis, we have devised a mass screening method involving serum pepsinogen measurement to identify subjects at high risk of gastric cancer. In 1991, we screened 4,647 workers (male: 4,113, female: 534, mean age: 49.0 years) at a Japanese company using this method. Out of 875 subjects (18.8%) with a serum pepsinogen I level of less than 50 micrograms/liter and a pepsinogen I/II ratio of less than 3.0, 676 subjects (14.5%) were selected for further investigation by endoscopy. This led to the detection of four subjects (0.086%) with gastric cancer (three in an early stage) and four subjects with adenoma. The cancer detection rate of this new screening method was comparable, and in some respects superior, to that of the traditional barium X-ray screening. Since the incidence of test-positive subjects was as low as 10% amongst subjects aged less than 40, this screening method appears to be especially useful for screening of younger generations. The new method is less expensive than the traditional barium X-ray and subjects experience little discomfort. Further, many serum samples can be quickly measured simultaneously. The results of this study have indicated that serum pepsinogen screening provides a valuable method for detecting gastric cancers. PMID- 8226284 TI - Edible plants containing naturally occurring carcinogens in Japan. PMID- 8226285 TI - Bone blood flow in the rat using arteriolar blockade: comparisons between labelled resin particles and microspheres. AB - Arteriolar blockade is probably the most widely used laboratory method for bone blood flow measurement. At first, cheap labelled cationic resin particles were used. These have largely been displaced by labelled microspheres, which have theoretical advantages but are expensive. This investigation compares measurements of bone blood flow made with the 2 different materials, using a reference artery method. Resin particles (1.4 mg per rat) were compared with 2 different doses of 85Sr-labelled microspheres (1 mg and 3 mg per rat). Whole bone blood flow from the 3 groups was not statistically different. Regional bone blood flow comparisons using resin particles and 1 mg dose microspheres showed no significant difference in epiphyses or metaphyses. In the resin group, marrow flow was significantly lower, and cortical flow elevated. It is suggested that these differences are caused by leaching of 59Fe to plasma, and methods of preventing this are discussed. Measured flow rates were significantly increased in the tibial marrow and cortex using 3 mg microspheres, compared with the 1 mg dose. This increase may reflect better sampling of the arteriolar beds. We conclude there is little advantage in using microspheres for bone blood flow measurement. When sufficient resin particles of appropriate size are injected and procedures are adopted to prevent leaching of isotope to plasma, the observed measurements are essentially identical. PMID- 8226286 TI - Morphological and histochemical characteristics of muscle fibre types in the flexor carpi radialis of the dog. AB - The canine flexor carpi radialis is subdivided into 2 portions. The radial portion which occupies one third of the cross-section is located caudomedially and is composed only of large type I fibres (43.53 +/- 0.6 microns minimum diameter) in levels close to the insertion. The cubital portion is located craniolaterally and contains fewer and smaller type I fibres (26.10 +/- 0.3 microns minimum diameter). It is hypothesised that the radial portion of the muscle, with its larger slow fibre population, contributes to forelimb postural function. The cubital portion, with its generally fast fibre population (IIA), may be more important in the muscle's recognised function (flexion of the carpal joint during locomotion). PMID- 8226287 TI - Novel articular cartilage structure in the South American opossum, Monodelphis domestica. AB - Articular cartilage provides smooth surfaces for low-friction, unrestricted movement of opposing skeletal elements. The surface topography of articular cartilage has been the subject of numerous studies and, with few exceptions, is considered to be smooth (at least at the light microscopic level). Some studies have reported 'humps' on the articular surface which have been related to underlying chondrocytes residing very close to the surface. Here we report on a highly nodular form of articular cartilage in the distal limb joints of the South American opossum, Monodelphis domestica. Unlike previous reports, these articular 'humps' are visible under a dissecting microscope. Each 'hump' or 'nodule' represents the surrounding matrix of single or sometimes paired rounded chondrocytes. Flattened chondrocytes normally associated with mammalian articular cartilage were absent from these joints. Interestingly, the articular cartilage of the more proximal limb joints such as the knee showed more typical features of articular cartilage including flattened superficial chondrocytes. PMID- 8226288 TI - Skeletal muscle fibre types in the dog. AB - Using a variety of histochemical methods we have investigated the mATPase reaction of skeletal muscle fibres in the dog. Types I, IIA, IIDog (peculiar to the dog) and IIC fibres were identified. The results reveal that the interpretation of the fibre type composition depends on the methods used. PMID- 8226289 TI - Apical-basal membrane polarity of membrane phosphatases in isolated capillary endothelium: alteration in ultrastructural localisation under culture conditions. AB - Capillaries from freshly isolated rat epididymal fat were subjected to protocols that allowed ultrastructural localisation of alkaline phosphatase and 5' nucleotidase. Alkaline phosphatase was almost entirely restricted to the capillary luminal membrane and vesicles associated with this membrane. 5' nucleotidase was localised on the basal or abluminal membrane and associated vesicles. Arterioles and occasional venules were also present in the cell isolates, and arteriole localisation of 5'-nucleotidase was identical to that in capillaries. In venules, 5'-nucleotidase often failed to exhibit a polarised distribution and was present on both membrane domains. In confluent cultured endothelial cells, 5'-nucleotidase was not expressed in a predominantly polarised arrangement. Alkaline phosphatase was found on apical surfaces and regions of lateral cell contact. The results of these studies show that capillary endothelial cells exhibit enzyme polarity of their surface membranes which is subject to change on introduction of the cells to tissue culture. PMID- 8226290 TI - Quantitative and spatial aspects of degenerative changes in rat soleus muscle after exercise of different durations. AB - The aim of the present study was to investigate the effect of exercise duration on the amount of histological change (degeneration and cellular response), the number of affected muscle fibres, the segment over which muscle fibres were affected and the spatial distribution of affected muscle fibres in the rat soleus muscle after submaximal uphill running. Male Wistar rats ran on a treadmill (10 degrees incline, 1840 m/h) for 1.5 or 2.5 h. Three days postexercise the left and right soleus muscles were removed. Serial transverse sections were cut from proximal to distal, so that individual fibres could be followed over their entire length. Histological changes were quantified morphologically. With increasing exercise duration, an increase in the histological changes was found, as were a greater number of affected fibres and an increase in the mean length over which fibres were affected. Beside occasional minor clustering, affected fibres were distributed randomly throughout the muscle. PMID- 8226291 TI - Vasomotor innervation of the skin of the hand: a contribution to the study of human anatomy. AB - The sympathetic vasomotor innervation of the skin of the human hand was studied in 47 subjects who underwent local anaesthetic block of ulnar, median or radial nerves at elbow or wrist levels. Areas of cutaneous anaesthesia were compared with cutaneous territories of paralytic vasodilatation delineated by infrared telethermography. It was found that: (1) during ulnar nerve block the area of vasodilatation matched the area of anaesthesia in all 15 cases; (2) median nerve block induced paralytic vasodilatation which, in 14 of 15 cases, matched the area of cutaneous anaesthesia in median territory, but also extended to the unanaesthetised lateral part of the dorsum of the hand; (3) no vasodilatation developed during radial nerve block in 17 of 18 cases, whereas areas of sympathetic sudomotor paralysis matched the area of radial sensory loss in all 5 subjects in whom sweating function was studied. It is concluded that: (1) the ulnar nerve supplies vasomotor fibres to its cutaneous sensory territory, no less and no more; (2) the median nerve normally provides supplementary vasomotor innervation to the skin of the radial aspect of the dorsum of the hand; (3) the radial nerve supplies sudomotor innervation for the lateral aspect of the dorsum of the hand, but (4) does not normally contribute vasomotor sympathetic fibres to the skin of the hand. PMID- 8226292 TI - Responses of dermal mast cells to injury. AB - The effect on dermal mast cell numbers and degranulation of making a partial thickness skin wound on the right flank of Wistar rats was studied immediately after operation and 0.5, 1, 2, 4, 8, 16, 24 and 72 h postoperatively. An equivalent area of intact dermis on the left flank was used as a control. In the injured dermis the mean number of detectable mast cells in the experimental group immediately after making the partial thickness wound was not significantly different from the control side (P > 0.25) but it later decreased, reaching its lowest value after 2 h and increasing from 16 h to 72 h postoperatively when the final assessment was made. The possibility that the reduction in mast cell number per unit area might be an artefact resulting from increased tissue volume due to oedema was investigated and disproved. The total number of dermal mast cells in equivalent areas of the intact left flank remained unchanged throughout this period. The percentage of degranulating mast cells started rising 0.5 h postoperatively, increased gradually to reach its highest value after 2 h, remained high up to 8 h postoperatively and then decreased to reach its lowest value after 72 h. The percentage of degranulating mast cells of the intact dermis of the left flank did not alter during this period. The lack of a significant change in the control groups shows either the absence of any systemic effect or that the technique used was not sensitive enough to detect it. PMID- 8226293 TI - Anatomy of Clara cell secretion: surface changes observed by scanning electron microscopy. AB - The microanatomical alterations of the surface of lung Clara cells were studied during secretion. Stimulation of Clara cells was induced in rats by chronic inflammation caused by a single intratracheal instillation of bleomycin performed 2.5 months before the animals were killed. Bleomycin treatment resulted in marked stimulation of secretion by the bronchiolar Clara cells, 31% of the Clara cells from the treated rats showing signs of active secretion whereas only 6.3% of Clara cells in control rats presented similar features. High-resolution views of lung airways were obtained by scanning electron microscopy of critical point dried tissue samples. The surface of Clara cells underwent several modifications associated with the secretory events. These alterations followed 3 major phases: (1) formation of a smooth apical dome made up of a large volume of cytoplasm; (2) progressive narrowing of this dome-like body at its base with the formation of a cap-like structure; (3) in toto release of the cytoplasmic cap-like body. In favourable views, thin pedicles linking the cap-like bodies to the remaining cytoplasm of the Clara cell were detected. In other instances, release of the cap like body occurred without the previous formation of stalks. Secretion of intracellular granules was observed in some cells before severance of the cap like body. It is concluded that: (1) the cap-like bodies are not artifactual features of Clara cells; (2) Clara cell secretion is both apocrine and merocrine, the former predominating; (3) chronic inflammation is associated with an increased formation and release of the secretory cap-like bodies by Clara cells. PMID- 8226294 TI - Early periosteal changes in translation-induced bone modelling. AB - This primarily ultrastructural study examines the effects of strain induced in the periosteum using an in vivo translation model with minimal internal bone strain. Caudal vertebrae (CV 7, 8, 9) from 4 d rats were threaded onto the arms of prestressed helical torsion springs and transplanted subcutaneously into 50 g hosts of the same inbred strain. After 7 d the appliances were activated in the experimental rats causing the bones to translate, i.e. to move through the soft tissues. Tissues for histology were obtained at this time (0) and at 1, 3, 5, 7, 10 and 14 d; for electron microscopy, experimental tissues were obtained at 0 time, 30 min, 1, 2, 6, 12, 18 and 24 h and at 0 time and 12 h for the controls. As the arms of the appliance move apart, traction on the enveloping soft tissues produces compression of the periosteum on the leading side and tension on the trailing side with resultant eccentric remodelling of the bones, generally opposite to the direction of movement. A rapid and differential structural response occurs, characterised by accelerated formation on the trailing side with the reverse on the leading, where changes are not as marked initially. Long thin trabeculae oriented in the line of tension form on the trailing side whereas the shaft on the leading side becomes thinner and flatter. Ultrastructural examination of the early stages shows that the fibrous periosteum is first affected, with alterations in collagen packing preceding cellular changes. The midzone shows the greatest change and events here presage those which finally occur at the bone surface and are reflected in altered osteoblastic activity. This study shows that translation-induced stress produces rapid morphological changes in the periosteum which, by acting as an integrated unit, has the capacity to modulate the adaptive bone modelling response. PMID- 8226295 TI - A quantitative study of the differentiation of microglial cells in the developing cerebral cortex in rats. AB - The distribution of various morphological forms of microglia stained with lectin in different regions of postnatal rat brain was examined in 2 planes of section, coronal and sagittal. In the coronal sections (mediolateral plane) taken at the level of the optic chiasma, the lectin-labelled cells were examined in the gustatory (GU), lateral somatosensory (LSS), medial somatosensory (MSS), motor (MO) and cingulate (CI) cortex. In the sagittal sections (rostrocaudal plane), the lectin-labelled cells in the medial orbital (MOA), motor (MO) and primary visual area (PVA) were studied. The cells in the motor area in coronal plane were further analysed with reference to their distribution in each of the laminae (layers). Based on the variation of their external morphology which represents different degrees of differentiation, all lectin-labelled microglial cells of the above-mentioned regions in newborn (P0) and rats aged 4 d (P4) and 8 d (P8) postnatum were classified and quantified. In the mediolateral plane of any given age group, the most differentiated ramified cells were located in GU except in P8 rats where the cells were also concentrated in CI. Of the 3 regions in the rostrocaudal plane the majority of the more differentiated ramified cells at P0 were found in the PVA but were the major cell type in MO in P4 and P8 rats. For the distribution of cells in MO, the most differentiated cells were located in the intermediate zones. It was concluded from this study that microglial cells in the developing cerebrum showed a gradient of differentiation in relation to different regions of the cerebral cortex but this appeared to vary with age. Thus in the mediolateral plane in P0 and P4 rats, the gradient extended from GU to CI in a lateromedial fashion but in P8, in the direction towards MSS from GU and CI. In the sagittal section, the gradient was directed caudorostrally in P0 rats. In P4 and P8 rats, however, the gradient was from MO to both poles (MOA, PVA). In the motor cortex, the gradient was from the intermediate zone towards the superficial and deep laminae. The gradient of differentiation of microglia may be related to the growth of the respective regions in the cerebral hemisphere but inherent genetic factors were also considered. PMID- 8226296 TI - Observations of the surface morphology of cultured granulated metrial gland cells. AB - The surface morphology of granulated metrial gland (GMG) cells which had migrated from cultured explants of mouse metrial gland tissue has been studied by scanning electron microscopy. Granulated metrial gland cells have a very characteristic 3 dimensional morphology with a particularly large cell body and the exposed surface of the cultured cells exhibits various combinations of thin microvillous processes and leaf-like processes. Some GMG cells have a relatively smooth surface. Many GMG cells have a leading lamellipodium characteristic of motile cells and long trailing filopodia. In mixed cell cultures it was possible to distinguish, on the basis of the combination of their size and surface features, GMG cells from macrophages, natural killer cells, lymphokine-activated cells and small lymphocytes derived from the peritoneal cavity or spleen. PMID- 8226297 TI - Microappendages on the atrioventricular valves of the guinea pig. AB - More microappendages are present on both surfaces of the guinea pig tricuspid valve than on the mitral valve. The distal edge of all valve surfaces has more microappendages than the remainder of the valve surface. These results suggest that microappendage number may be the result of haemodynamic forces on the valve surface: the greater the force the smaller the number of microappendages. The function of microappendages on the surfaces of the heart valves is unknown, but two important roles have been suggested, namely to increase the surface area of the endothelial cells and to participate in the metabolism of these cells. PMID- 8226298 TI - Morphology of the most common form of protostylid on human lower molars. AB - Scanning electron microscopy of the enamel surface and enamel structure in the protostylid area classified as a surface irregularity was performed on 26 human lower third molars. This protostylid class can be regarded as the onset of a cusp formation, while the protostylid pits should be considered to be fissures between a protoconid and a poorly expressed protostylid. PMID- 8226299 TI - Innervation of the anteromedial muscle bundles of the gluteus medius. AB - The independent anteromedial muscle bundles situated lateral to the nerve to tensor fasciae latae were examined in 13 human pelvic halves. Each of these independent muscle layers was shown not to be an independent muscle but part of gluteus medius. On the basis of its innervation, tensor fasciae latae is closely related to the most anteromedial muscle layer of gluteus medius. From their pattern of innervation, these anteromedial bundles may be considered as a continuation of gluteus medius rather than as completely independent. Furthermore, tensor fasciae latae is closely related to the most anteromedial of these bundles. PMID- 8226300 TI - A rare anomaly of abductor digiti minimi. AB - Two cases with anomalous fascicles in abductor digiti minimi, noted in the course of dissecting 62 adult postmortem forearms, are described. Both fascicles arose from the flexor retinaculum and the antebrachial fascia; one was inserted into abductor digiti minimi and the other on the proximal phalanx of the 5th finger. The anomalous muscles crossed the ulnar nerve and in 1 case also the median nerve. In the 2nd case the palmar nerve to the 5th finger was seen to penetrate the anomalous muscle. The ontogeny, morphology and clinical significance of this anomaly are discussed in relation to previously described anomalies of the hypothenar muscles. PMID- 8226301 TI - An anatomical study of the deep transverse metacarpal ligament. AB - The anatomy of the deep transverse metacarpal ligament was examined in cadaveric dissections. Previously unreported findings were transverse muscular slips from abductor digiti minimi that inserted into the medial border of the ligament and, in 1 case, an extension from the medial half of the tendon of extensor carpi ulnaris that inserted into the medial aspect of the ligament. Applied anatomy in relation to the dynamics of the transverse metacarpal arch is reviewed. PMID- 8226302 TI - Effect of spermatocele fluid on growth of human prostatic cells in culture. AB - The present study was carried out to investigate whether testicular fluid derived from a spermatocele contains substance(s) that promote the growth of human prostatic cells in culture. Human spermatocele fluid was centrifuged to sediment spermatozoa. The supernatant was then added to cultures of human prostatic stromal or epithelial cells that were isolated from surgical specimens of benign prostatic hyperplasia. Addition of spermatocele fluid in quantities of 1 microgram/ml of protein resulted in a significant increase in the number of both prostatic stromal and epithelial cells at the end of a 6-day culture period. Human serum at equivalent protein concentrations in the culture medium had no stimulatory effect. At least two separate growth-promoting factors were found in spermatocele fluid, one for stromal cells and one for epithelial cells. The mitogen for stromal cells was heat labile and persisted after treatment with activated charcoal. The factor for epithelial cells was heat stable but was removed by charcoal treatment. These observations are consistent with the concept that the human testis secretes nonandrogenic substances that can promote prostatic growth. PMID- 8226303 TI - Effects of epidermal growth factor on human sperm cell function. AB - The effects of human recombinant epidermal growth factor (EGF) on the fertilizing capacity of human sperm were investigated. At lower concentrations (0.1-10 nM) EGF did not significantly (P > 0.05) affect human sperm penetration of zona-free hamster oocytes (SPA). At higher concentrations (25-100 nM), EGF significantly (P = 0.01 to < 0.001) decreased the human sperm penetration rates in SPA. At higher concentrations (> or = 25 nM), EGF also significantly inhibited the spontaneous as well as calcium ionophore-induced acrosome reaction and release of acrosin from human sperm. There was no effect of EGF on percent sperm motility, but at higher concentrations (> or = 25 nM) it significantly affected various sperm motility characteristics especially velocity and amplitude of lateral head displacement. EGF was detected by radioimmunoassay as well as radioreceptor assay in seminal plasma of fertile men. However, there were no statistical differences between the levels of EGF or EGF/transforming growth factor (TGF-alpha) in seminal plasma of fertile, infertile, and immunoinfertile men. Also, there was no significant correlation of the EGF or EGF/TGF-alpha levels with total sperm number, sperm motility characteristics, and penetration rates in SPA in these patients. These results indicate that EGF has either no effect or an inhibitory/negative effect on the human sperm cell capacitation and/or acrosome reaction, especially at higher concentrations (> 25 nM). PMID- 8226304 TI - Alteration of mRNA transcript levels of rat testicular cells following procarbazine administration. AB - The present study examined the effects of a single 400-mg/kg dose of procarbazine upon various testicular mRNA transcript levels in adult Sprague-Dawley rats. Northern blot hybridization was employed to measure the steady-state mRNA levels of proteins specific for Sertoli cells (adrogen-binding protein [ABP] and transferrin) and spermatids (protamine 1 and transitional protein 2). In addition, mRNA transcript levels were determined for germ cell-specific hemiferrin and insulin-like growth factor-1 (IGF-1), which in adult rats is present predominantly in primary spermatocytes. Furthermore, the chronology of the effects of procarbazine upon spermatogonial populations was examined in whole mounts of seminiferous tubules. The effect of procarbazine upon spermatogenesis was first noted among mature A3, young A4, and B spermatogonia 48-72 hours after drug administration. These results indicate that A2 through intermediate spermatogonia were most susceptible to procarbazine. In addition, degeneration of spermatocytes and young spermatids was evident by 5 days. Northern blot analysis of testicular poly (A)+ RNA revealed that the steady-state levels of mRNA transcripts of the spermatid nuclear proteins (protamine 1, 700 bp; and transitional protein 2, 580 bp) remained relatively unchanged for 5 days after procarbazine administration but were significantly decreased by more than 70% by 7 days. On the other hand, the steady-state mRNA levels for the Sertoli cell proteins ABP (1.7 kb) and transferrin (2.7 kb) were decreased by 45% and 50% (P < 0.05), respectively, 2 days after procarbazine administration. Significant suppression of ABP mRNA levels persisted through day 5, with partial recovery noted on day 7.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226305 TI - Protection by gonadal steroid hormones against procarbazine-induced damage to spermatogenic function in LBNF1 hybrid rats. AB - Some anti-cancer drugs, such as procarbazine (PCZ), are associated with irreversible damage to spermatogenic function and cause sterility in men. In the present study, protection of spermatogenesis by gonadal steroid hormones during PCZ treatment was investigated in male rats. LBNF1 hybrid rats were chosen for these studies because the response of the testis to PCZ was more uniform than in a stock of Sprague-Dawley outbred rats. Mature male LBNF1 rats were subcutaneously implanted with cholesterol (C)-, testosterone (T)-, and/or estradiol-17 beta (E)-containing capsules, and 5-10 weeks later given one or four weekly intraperitoneal injections of PCZ at a dose of 200-250 mg/kg of body weight. Hormone capsules were removed 24 hours after the last PCZ injection and the animals were killed 10-12 weeks later. Testicular weights, sonication resistant sperm head counts, and quantitative testicular histology revealed protection of the spermatogenic epithelium from PCZ toxicity by gonadal steroid hormones when compared with the C controls. Protection was observed against both single and multiple injections of PCZ. A combination of T and E capsules provided better protection than T alone. This model system is suitable for studies of the mechanism of protection of spermatogenesis from chemotherapy-induced damage. PMID- 8226306 TI - Comparison of effects of 0.5 and 3.0 Gy X-irradiation on lipid peroxidation and antioxidant enzyme function in rat testis and liver. AB - The prooxidant effect of X-irradiation on rat testis and liver tissue was studied with doses of 0.5 and 3.0 Gy; the latter dose kills the proliferating spermatogonia and causes a maturation-depletion process in the germ cells. The level of lipid peroxidation, measured by the formation of diene conjugates and thiobarbituric acid-reactive substances (TBARS) and the activities of the antioxidant enzymes were determined 0.5 hours, 1 day, 7 days, and 31 days after the exposure. In the liver, increased levels of diene conjugation (+36%, P < 0.05) in the group of 3.0 Gy at 0.5 hours indicated increased lipid peroxidation. At the same time, TBARS were increased (+25%, P < 0.05) in the group of 0.5 Gy, but not in the 3.0-Gy group. In the testis, diene conjugation was not determined at 0.5 hours postirradiation, and at day 1 it was at the control level. The level of TBARS in the testis was below control (-11%, P < 0.01) in the 3.0-Gy group at day 1. At day 31 after 3.0 Gy in the testis, an increase in the amount of conjugated dienes (+24%, P < 0.01) was observed in parallel with a decreased level of TBARS (-15%, P < 0.01). The activity of superoxide dismutase (SOD) was decreased in the testis at 0.5 hours postirradiation (-28%, P < 0.05, and -29%, P < 0.05, in the groups of 0.5 and 3.0 Gy), whereafter it returned to normal by day 7. In the liver, such inactivation of SOD was not observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226307 TI - Antifertility effects of neem (Azadirachta indica) oil in male rats by single intra-vas administration: an alternate approach to vasectomy. AB - An alternate approach to vasectomy for long-term male contraception following a single intra-vas application of a traditional plant (Azadirachta indica) product having immunomodulatory properties is described. Male Wistar rats of proven fertility were given a single dose (50 microliters) of neem oil in the lumen of the vas deferens on each side; control animals received the same volume of peanut oil. Animals were put on continuous mating 4 weeks after the treatment, with females of proven fertility. While the control animals impregnated the female partners, all males treated with neem oil remained infertile throughout the 8 months of observation period. Epididymal and vas histology were normal without any inflammatory changes or obstruction. The intra-vas administration of neem oil resulted in a block of spermatogenesis without affecting testosterone production; the seminiferous tubules, although reduced in diameter, appeared normal and contained mostly early spermatogenic cells. No anti-sperm antibody could be detected in the serum. Unilateral administration of neem oil in the vas resulted in a significant reduction of testicular size and spermatogenic block only on the side of application; the draining lymph node cells of the treated side also showed enhanced proliferative response to in vitro mitogen challenge. These results indicate that the testicular effects following intra-vas application of neem oil may possibly be mediated by a local immune mechanism. PMID- 8226308 TI - Studies on sperm chromatin structure alterations and cytogenetic damage of mouse sperm following in vitro incubation. Studies on in vitro-incubated mouse sperm. AB - Mouse epididymal sperm incubated in Tyrode's T6 fertilization media were analyzed over time for chromosome damage by two methods. First, cytogenetic analysis was done on paternal pronuclei metaphase chromosomes. After 6 hours incubation 11% of the cells demonstrated chromosome structural abnormalities. Secondly, sperm nuclei were measured by the sperm chromatin structure assay, which is a measure of the susceptibility of sperm DNA to the nuclei demonstrated an increased susceptibility to DNA denaturation, reaching near 100% by 48 hours. Changes in chromatin structure at the molecular level may lead to chromosome breaks seen in pronuclear chromosomes. PMID- 8226309 TI - Use of the Florida Sexual History Questionnaire to differentiate primary organic from primary psychogenic impotence. AB - A recent study demonstrated the Florida Sexual History Questionnaire (FSHQ) is reliable and valid when used to discriminate impotent from nonimpotent men. This study examined the ability of the FSHQ to discriminate between men with primary psychogenic and primary organic impotence in a sample of impotent diabetics and a larger group of impotent men. The hit rate in the diabetic sample was 81% using an eight-item discriminant function, and 70% using the total score. Hit rates in the larger sample were significant but lower (73% and 66%). The results suggest that the FSHQ has concurrent validity in terms of discriminating primary organic from primary psychogenic impotence. PMID- 8226310 TI - WB2838 [3-chloro-4-(2-amino-3-chlorophenyl)-pyrrole]: non-steroidal androgen receptor antagonist produced by a Pseudomonas. AB - In the course of our search for non-steroidal androgen-receptor antagonists of microbial origin, Pseudomonas sp. No. 2838 was found to produce an inhibitor of androgen binding to its receptor. This compound, named WB2838, was isolated and identified as 3-chloro-4-(2-amino-3-chlorophenyl)-pyrrole. The IC50 value of WB2838 for partially purified rat prostate cytosol receptor was 8.0 x 10(-7) M. However, the IC50 value of WB2838 against estrogen-receptor binding was about 90 fold greater than that against androgen-receptor binding. WB2838 inhibited the growth of androgen-responsive mouse mammary carcinoma SC-3 cells in the presence of 10(-8) M testosterone at IC50 value of 4.1 x 10(-7) M. This inhibition was reversed by adding 10(-5) M testosterone to the culture medium. WB2838 also showed the inhibitory activity against the growth of the ventral prostate induced by testosterone propionate in castrated immature rats. Therefore, it was concluded that WB2838 was a non-steroidal androgen-receptor antagonist. PMID- 8226311 TI - Oligomycin F, a new immunosuppressive homologue of oligomycin A. AB - Oligomycin F, a new homologue of oligomycin A (1), was isolated from a Streptomyces species and structure 2 was deduced by NMR methods. Compound 2 is highly active against plant pathogenic fungi and is an extremely potent suppressive agent for various immunological systems. PMID- 8226312 TI - Dephostatin, a novel protein tyrosine phosphatase inhibitor produced by Streptomyces. I. Taxonomy, isolation, and characterization. AB - A novel inhibitor of protein tyrosine phosphatase, dephostatin, was isolated from the culture broth of a strain of Streptomyces. The active principle was extracted from the broth filtrate with ethyl acetate and purified by silica gel chromatography and by HPLC. Dephostatin inhibited protein tyrosine phosphatase prepared from a human neoplastic T-cell line with an IC50 at 7.7 microM. The inhibitory pattern of dephostatin was competitive against the substrate. Dephostatin inhibited the growth of Jurkat cells. PMID- 8226313 TI - Isolation, structures, and antifungal activities of new aureobasidins. AB - Aureobasidins are a group of cyclic depsipeptides with antifungal activity and are produced by Aureobasidium pullulans. Aureobasidins are composed of eight amino acids and one hydroxy acid such as 2-hydroxy-3-methylpentanoic acid (Hmp), and highly lipophilic. Five new aureobasidins, S1, S2a, S2b, S3 and S4, which have higher hydrophilicity in reversed phase HPLC than the known aureobasidins A R, were discovered in a fermentation broth of A. pullulans R106 by means of on line liquid chromatography/mass spectrometry with electrospray ionization. We identified the structures of the compounds and studied their antifungal activities. Three of the new aureobasidins, S2b, S3 and S4, which have hydroxylated Hmp as the hydroxy acid, were highly active against Candida spp. and Cryptococcus neoformans. PMID- 8226314 TI - Further novel metabolites of the paraherquamide family. AB - Four novel metabolites of a Penicillium strain, IMI 332995, which has previously been reported to produce paraherquamide and a number of related metabolites, are herein described. VM55596 is the first N-oxide to be found in this family of compounds. Unusual oxidative substitution is also seen in VM55597. VM55599 appears to be the first documented example of the hexacyclic indole species that have long been postulated as biosynthetic precursors of metabolites of the brevianamide, paraherquamide and marcfortine families. PMID- 8226315 TI - Milbemycins alpha 11, alpha 12, alpha 13, alpha 14 and alpha 15: a new family of milbemycins from Streptomyces hygroscopicus subsp. aureolacrimosus. Taxonomy, fermentation, isolation, structure elucidation and biological properties. AB - Streptomyces hygroscopicus subsp. aureolacrimosus SANK 60286 produces a new family of milbemycins, named milbemycins alpha 11, alpha 12 [corrected], alpha 13, alpha 14 and alpha 15, together with other milbemycins. Their structures are 3-methyl-2-butenoyloxy and 3-methylbutyroyloxy derivatives at C-4a of milbemycins A3 and A4, or 3-methyl-2-pentenoyloxy derivative at C-4a of milbemycin A3, respectively. Milbemycin alpha 14, 3-methyl-2-butenoyloxy derivative, especially possesses a potent acaricidal activity. PMID- 8226316 TI - A non-enzymatic reaction in the late biosynthesis of the decarestrictine family. AB - In the late biosynthesis of the decarestrictines, inhibitors of the de novo formation of cholesterol isolated from the culture broth of Penicillium simplicissimum (FH-A 6090), a common pentaketide precursor undergoes post polyketide modifications leading to the different members of the decarestrictine family. Besides subsequent enzymatically catalyzed reactions an unexpected non enzymatic conversion was found to be the key step in the biosynthetic sequence. Under acidic conditions during fermentation the decarestrictines A1 and A2 (3 and 4) are converted into the main product decarestrictine D (2) and the new decarestrictines N (5) and O (6), whose physico-chemical data are reported. Mechanistic aspects of the non-enzymatic reaction as well as a more detailed picture of the biosynthetic relationships of the decarestrictine family are described. By the application of pH-static fermentations these results were successfully used to manipulate the secondary metabolite pattern of strain FH-A 6090 directing the fermentation process to produce desired members of the decarestrictine family. PMID- 8226317 TI - The squalestatins, novel inhibitors of squalene synthase produced by a species of Phoma. IV. Preparation of fluorinated squalestatins by directed biosynthesis. AB - Feeding of fluorinated benzoic acids to fermentations of a Phoma sp. resulted in the biosynthesis of a series of novel fluorinated squalestatins. The feeding studies, isolation, structural elucidation and biological activities of these compounds are reported. PMID- 8226318 TI - Delaminomycins, novel extracellular matrix receptor antagonists. V. Biosynthesis. AB - The biosynthesis of delaminomycin A, produced by Streptomyces albulus MJ202-72F3, was investigated by feeding 13C-labeled compounds followed by 13C NMR analyses. The results indicate that delaminomycin A is derived from six acetate units, five propionate units and one glycine unit. PMID- 8226319 TI - Biological properties of aureobasidin A, a cyclic depsipeptide antifungal antibiotic. AB - Aureobasidin A (AbA) is a novel cyclic depsipeptide antifungal antibiotic. The antifungal activity of AbA was studied in vitro and in vivo in comparison with clinically effective antifungal agents, amphotericin B and fluconazole. AbA was highly active in vitro against many pathogenic fungi, including Candida albicans, Cryptococcus neoformans, Blastomyces dermatitidis and Histoplasma capsulatum. The activity was superior to amphotericin B in most cases. AbA exhibited fungicidal action toward growing cultures of C. albicans. It was highly tolerated by mice and showed good efficacy in the treatment of murine systemic candidiasis when given orally or subcutaneously. AbA's fungicidal action in mice with candidiasis was more effective than fluconazole and amphotericin B. PMID- 8226320 TI - CL307-24, a new antibiotic complex from Saccharopolyspora aurantiaca sp. nov. II. Physico-chemical and biological properties. AB - CL307-24I, the main component of the CL307-24 complex produced by Saccharopolyspora aurantiaca sp. nov., was found to be a potent inhibitor of yeast mitochondrial ATPase. CL307-24I displayed a high degree of activity towards some coryneform bacteria and also has been shown to possess an insecticidal activity. Its biological and physico-chemical properties clearly distinguish it from previously known ATPase inhibitors. PMID- 8226321 TI - Synthesis and structure-activity relationships of new dimeric mitomycin derivatives; 7-N,7'-N'-bis(omega-thioalkyl)dimitomycins. AB - The reaction between mitomycin A (1) and cysteamine afforded 7-N,7'-N'-bis(2 thioethyl)dimitomycin C (7), 7-N-[2-[(2-aminoethyl)dithio]ethyl]mitomycin C (8), and 7-methoxy mitosenes (10, 11). The structures of 7 and 8 were elucidated on the basis of spectroscopy and reactions between 1 and 8, and 1 and cystamine. The observation of the time course for the reaction revealed the mechanism of the formation of 7 and 8. The rapid oxidation of cysteamine by the quinone of 1 gave cystamine, which was trapped by 1 to give 8, and 8 was additionally reacted with 1 to give 7. Since 7 showed significant antitumor activities, related 7-N,7'-N' bis(omega-thioalkyl)dimitomycins were synthesized. They also showed remarkable antitumor activities against HeLa-S3 in vitro, sarcoma 180 (sc-ip), leukemia P388 (ip-ip) in vivo. In these evaluations, compound 7 demonstrated unique potency. PMID- 8226322 TI - Structure-antitumor activity relationship of semi-synthetic spicamycin analogues. AB - Spicamycin, a nucleoside antibiotic containing fatty acids with a variety of chain lengths (C12-C18), showed potent antitumor activity against human gastric cancer SC-9 and human breast cancer MX-1 in a xenograft model. We have made several semi-synthetic spicamycin analogues (SPMs) which differed in the chain length of the fatty acid moiety, and examined their structure-antitumor activity relationship. The cytotoxic activities of SPMs depended on the chain length of the fatty acid moiety, with dodecanoyl, tetradecanoyl, hexadecanoyl and icosanoyl analogues (SPM VIII, SPM X, SPM XII and SPM XVI) exhibiting the most potent cytotoxic activity against P388 murine leukemia cells. SPM VIII showed the most activity against SC-9 in the human tumor xenograft model with the highest therapeutic index among SPMs. The antitumor activity of SPM VIII was superior to that of mitomycin C. PMID- 8226323 TI - Synthesis and antifungal activity of pradimicin derivatives. Modifications on the aglycone part. AB - Synthesis and antifungal activity of pradimicin analogs modified on the aglycone part is described. Upon modification studies at various sites of the aglycone part using pradimicin A (PRM A), C-11 position was found to be the sole site to be modified without loosing antifungal activity. Further modification studies at C-11 position were carried out with 11-OH derivative of pradimicin T1 (PRM T1) because of its easy availability. Among the compounds prepared, 11-demethoxy derivative of PRM A (12) and 11-O-ethyl (13) and 11-O-fluoroethyl (14) derivatives of PRM T1 showed promising antifungal activity comparable to that of PRM A. PMID- 8226324 TI - Synthesis and biological activity of C-3' ortho dihydroxyphthalimido cephalosporins. AB - A series of C-3' ortho dihydroxyphthalimido cephalosporins 3-7 has been prepared by reaction of C-3' aminomethyl cephalosporin 41 with the corresponding N carboethoxyphthalimides 23-25, 37, 38. These new cephalosporins exhibit excellent in vitro Gram-negative activities, including Pseudomonas aeruginosa, excellent beta-lactamases stability and pharmacokinetics equivalent or better than ceftriaxone. PMID- 8226325 TI - Cysfluoretin, a new inhibitor of glutathione S-transferase, produced by Streptomyces sp. MI384-DF12. PMID- 8226326 TI - The amino acid sequence of actinoxanthin apoprotein deduced from the base sequence of the gene. PMID- 8226327 TI - Studies on the biosynthesis of fosfomycin. V. Cloning of genes for fosfomycin biosynthesis. PMID- 8226328 TI - Localization of preproenkephalin mRNA-expressing cells in rat auditory brainstem with in situ hybridization. AB - Hair cells and auditory nerve dendrites in the inner ear are innervated by pontine neurons that have been demonstrated by immunochemical techniques to contain several neurotransmitters, including acetylcholine and the opioid peptide enkephalins and dynorphins. The functions of these nerve fibers are not known, but may involve modifying auditory sensitivity to low intensity stimuli. In the guinea pig the opioid pathways originate in the lateral superior olivary region. A recent study in the gerbil has reported cells expressing preproenkephalin mRNA present only in the ventral nucleus of the trapezoid body, and not in the superior olivary region. In the present study, a non-radioisotopically labeled in situ hybridization method was used to identify cells expressing mRNA coding for preproenkephalin in rat pontine neurons, specifically in the ventral nucleus of the trapezoid body. These cells may represent an enkephalin-containing medial olivocochlear system in the rat, the origin of the lateral system in the rat that differs markedly from the better-studied guinea pig and cat, or a non olivocochlear enkephalin-containing system. PMID- 8226329 TI - High affinity aldosterone binding sites (type I receptors) in the mammalian inner ear. AB - The presence of aldosterone (Type 1) binding sites in the mammalian inner ear has been previously suggested by an increase in inner ear Na, K-ATPase ouabain binding sites in response to the administration of aldosterone in vivo (Pitovski et al., 1993). Type I binding sites have now been identified and characterized in the lateral wall of the basal turn of the cochlea and in the ampullae of the semicircular canals of the guinea pig. In the presence of RU 28362, which blocks low-affinity binding of the labeled hormone to Type II sites, [3H]-aldosterone binds to a single class of high-affinity (Type I) sites with Kd values of 34.7 nM in lateral wall of the basal turn of the cochlea and 31.3 nM in the ampullae of the semicircular canals. Bmax is 17.1 fmol/mg dry tissue for the cochlear sample and 17.4 fmol/mg dry tissue for the ampullae, comparable to reported values in renal tissue (17-31 fmol/mg protein). Thus, the results of receptor-binding experimental protocols with [3H]-aldosterone clearly suggest that these inner ear tissues are a target site of mineralocorticoid action. PMID- 8226330 TI - The Na+/H+ exchanger in transitional cells of the inner ear. AB - The cytosolic pH (pHi) of transitional cells from the ampulla of the gerbil was measured micro-fluorometrically with the pH-sensitive dye 2',7'-bicarboxyethyl 5(6)-carboxyfluorescein (BCECF) to assess the possible contribution of a Na+/H+ exchanger to the regulation of pHi. All experiments were conducted in virtually HCO3(-)-free solutions. Under control conditions, pHi was 7.19 and addition of 10(-5) M ethylisopropylamiloride (EIPA), a blocker of Na+/H+ exchange, caused a small but significant acidification of pHi. A transient exposure to 21.4 mM NH4Cl caused a rapid cytosolic alkalinization followed by a brisk acidification and prompt recovery of pHi to its control value. The cytosolic buffer capacity (Bi) was determined in the absence of Na+ from changes in pHi which were elicited by [NH4+] steps. Bi was 4.7 mM/pH at pHi 7.19 and varied with pHi. The initial net proton flux JH, representative of Na+/H+ exchange activity, was calculated from the product of the initial rate of alkalinization after an NH4(+)-prepulse and Bi at the corresponding pHi. JH was dependent on the extracellular Na+ with an apparent Km of 64 mM, sensitive to the cytosolic [H+] with an apparent Km of 2.7 * 10(-7) M (i.e. pH 6.6), and inhibited by EIPA with an IC50 of 5 * 10(-7) M. These data suggest that transitional cells contain in the basolateral membrane a Na+/H+ exchanger of the amiloride-sensitive subtype. PMID- 8226331 TI - Aspects of spontaneous otoacoustic emissions in healthy newborns. AB - Spontaneous otoacoustic emissions (SOAEs) are pure-tone like signals, spontaneously present in the ear canal. In normal adult ears the prevalence of SOAEs is reported to be 30-70%, probably depending on the noise floor of the recordings. In infant studies, results on the SOAE prevalence are rare. SOAEs as well as evoked otoacoustic emissions (EOAEs) were recorded in healthy newborns. Their ages varied between 1 and 10 days. The recordings were done with commercially available equipment in a separate not sound treated room of the obstetric department. The prevalence of SOAEs was 78%, which is higher than previously reported for adults as well as healthy newborns. The prevalence was not significantly different between left and right ears, or genders. The number of emissions per emitting ear amounted on average 5.5. The median number of SOAEs in boys (3.3) is significantly lower than in girls (4.6). The SOAE levels were between -2 and 42 dB SPL. The mean level per emitting ear was 8.0 dB SPL and not significantly different between right and left ears or genders. However, the level of the strongest emission per emitting ear was significantly higher for right than for left ears. In contrast with adults most of the emissions (70%) are at frequencies above 2 kHz. Comparing the levels of the EOAEs between ears with and without SOAEs we found a statistically significant higher EOAE level in ears with SOAEs. This supports our previous hypothesis that the higher EOAE level found in healthy newborns is partly due to the more frequent presence of stronger SOAEs in healthy newborns.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226332 TI - Two types of chloride channel in the basolateral membrane of vestibular dark cells. AB - Transepithelial and cell membrane potential measurements have suggested that the basolateral membrane of gerbil vestibular dark cells contains Cl- conductive pathways. We used the patch clamp technique to search this membrane for Cl- conductive channels which could account for the macroscopic observations. Two types of Cl- channel were found in both cell-attached and excised membrane patches. One type was found with an incidence of 19% and had a single-channel conductance of 95 +/- 1 pS (N = 20) in symmetrical Cl- solutions. The other type was found with an incidence of 3% and had a large single-channel conductance of 360 +/- 11 pS (N = 12) in symmetrical Cl- solutions (LC-type Cl- channel). Both types of Cl- channel had linear current-voltage relations and at least 2 substates. In asymmetrical Cl- solutions (gluconate substitution) the current voltage relations fit the Goldman-Hodgkin-Katz current equation for Cl-. Neither channel was blocked by Zn2+, NPPB, DIDS, DNDS or quinine. The 95 pS channel exhibited a spontaneous 'rundown' of its activity within 1 to 10 min after being excised. This rundown was not reversed by the catalytic subunit of protein kinase A. Channel activity was not dependent on the presence of cytosolic Ca2+ nor markedly altered by variations in cytosolic pH between 6.5 and 8.0. The two Cl- channels were distinguished by the membrane voltage ranges in which they were active and by their anion selectivity. The open probability of the 95 pS channel was insensitive to voltage and the anions NO3-, I- and Br- were only half as permeable as Cl-. By contrast, the LC-type Cl- channel was mostly active between about +/- 30 mV and equally permeable to NO3-, I-, Br- and Cl-. The 95 pS Cl- channel may account for the observed transepithelial and intracellular voltage responses to Cl- concentration steps and provide the path for the recirculation of Cl- across the basolateral membrane. The LC-type Cl- channel shows the same lack of anion discrimination as the anion pathway activated during hyposmotic challenge. PMID- 8226333 TI - Effects of atmospheric pressure variation on spontaneous, transiently evoked, and distortion product otoacoustic emissions in normal human ears. AB - The effects of atmospheric pressure changes on the frequency and amplitude of spontaneous (SOAEs), transiently evoked (TEOAEs) and distortion product (DPOAEs) otoacoustic emissions in normally hearing humans were compared. The purpose was to determine if the transmission of each form of OAE was influenced differently by the middle ear. Sixty-one subjects were tested in a pressure chamber. Twenty seven SOAEs with a frequency range between 535 to 4729 Hz from 21 subjects were examined. Transiently evoked OAEs were studied in 20 subjects using clicks and tone-bursts at 0.5, 1, 2, 3, and 4 kHz. Distortion-product OAEs were generated at seven geometric mean frequencies between 1 and 8 kHz in another 20 subjects. Spontaneous OAEs were examined by applying atmospheric pressure up to 9 kPa and down to -2.5 kPa, for the measurement of TEOAEs and DPOAEs the pressure was varied from 0 kPa up to 8 kPa. In spite of large interindividual differences, results suggest that the influence of pressure on the three OAEs is frequency specific. The frequency and amplitude change of SOAEs, the modification of the amplitude and spectra of TEOAEs, and the amplitude change of DPOAEs are more influenced by changes in middle ear pressure below 4 kHz than are OAEs in the range at 4 kHz and above. PMID- 8226334 TI - Detection of HSP 72 synthesis after acoustic overstimulation in rat cochlea. AB - The purpose of this study was to determine if high intensity acoustic stimulation would induce HSP 72 in rat cochlea. The animals were exposed to 110 dB SPL broad band noise for 1.5 h and sacrificed 4, 6 and 8 h after stimulation. Immunocytochemistry and western blotting were used to detect the expression of HSP 72 in the cochlear tissues. Western blots showed an intense 72 kD band in the noise exposed animals compared to a very light band in non-stimulated control animals. Immunocytochemical results in the cochlea revealed noise induced HSP 72 immunoreactive staining of outer hair cells. Only a few immunoreactive stained inner hair cells were seen and spiral ganglion cells were not stained. These results indicate that acoustic overstimulation can induce the expression of HSP 72 in outer hair cells of the rat cochlea. HSP 72 may serve as a marker for cellular stress and potential damage and may be involved in protection from insult. PMID- 8226335 TI - Recovery of CAP threshold and amplitude in chickens following kanamycin ototoxicity. AB - Chickens were given a dose of kanamycin (400 mg/kg/d x 10 d) which destroyed hair cells over the basal 37-58% of the basilar papilla. Afterwards, the threshold and amplitude of the compound action potential were measured at recovery times ranging from 2 days to 10-20 weeks post-kanamycin treatment. At 2 days post treatment, the thresholds at 1000, 2000 and 4000 Hz were elevated 40-60 dB while the thresholds at 250 and 500 Hz were elevated only 25 dB. By 10-20 weeks post treatment, the threshold at 250 and 500 Hz had completely recovered whereas a residual threshold shift of 5 dB to 25 dB was present between 1000 to 4000 Hz. The maximum amplitude of the compound action potential was also reduced by more than 60% at all frequencies at 2 days post-treatment; however by 10-20 weeks post treatment, the amplitude of the compound action potential had completely recovered at 500, 1000 and 2000 Hz. By contrast, the amplitude of the compound action potential at 4000 Hz was still reduced by more than 50% of its normal value 10-20 weeks post-treatment. The results of the present study indicate that the time course of recovery of the compound action potential is extremely slow and may lag behind the regeneration of hair cells by many weeks. The permanent deficits observed at the high frequencies could conceivably be due to functional deficits in regenerated hair cells, their afferent synapses or the loss of cochlear ganglion cells. PMID- 8226336 TI - Discharge-rate dependence of refractory behavior of cat auditory-nerve fibers. AB - A descriptive model for auditory-nerve (AN) refractory periods is described. The model assumes that interspike intervals consist of a constant-length absolute refractory period (ARP), followed by random-length relative refractory period (RRP) and a random-length waiting time to the next spike. Both the RRP and waiting time are exponentially distributed. This model fits AN hazard functions sufficiently well to provide estimates of the ARP and RRP durations for each fiber. The ARP is found to be constant, independent of discharge rate, with mean value between 0.56 and 0.86 ms in data from 7 experiments. The RRP decreases in duration as discharge rate increases; RRP mean length is less than 2 ms in most cases. There is an additional, slow component of recovery, lasting 20-40 ms, which is not modeled. The variation in RRP with discharge rate is shown to be capable of accounting for the deviation of AN regularity from that predicted for a Poisson process. Finally, properties of peaks seen in hazard functions just at the end of the ARP are described; these peaks are not included in the model, but are shown to be especially prominent in low and medium spontaneous rate fibers. PMID- 8226337 TI - The functional morphology of stereociliary bundles on turtle cochlear hair cells. AB - The stereociliary bundles of hair cells from the basilar papilla of the red-eared turtle were examined with transmission and high resolution scanning electron microscopy to provide a description of their morphology, orientation and inter ciliary connections for comparison with physiological observations. Bundles on hair cells in the basilar membrane region are of a uniform shape and orientation, but bundles on the apical and basal limbus are distinct in having elongated kinocilia more than twice the length of the tallest stereocilia. Bundles in the basilar membrane region show a roughly two-fold increase in height from 5 to 9 microns from base to apex. Electrical recordings from isolated hair cells indicate that the bundle height is inversely proportional to the cell's characteristic frequency. It is argued that the change in dimensions is insufficient to contribute significantly to the cochlea's frequency selectivity. The cytoplasm adjacent to the kinocilium is filled with microtubules and large vesicles, and there are coated pits in the apical membrane which, it is suggested, may be indicative of rapid turnover of the membrane in this region. PMID- 8226338 TI - Interrelation of spontaneous and evoked otoacoustic emissions. AB - The interrelation of spontaneous otoacoustic emissions (SOAEs) that can be synchronised by acoustic stimuli and transiently evoked otoacoustic emissions (TEOAEs) was studied in different experiments in normal ears. Click evoked TEOAEs mixed with synchronised SOAEs were investigated in time windows of 102.4 ms. Frequency spectra were obtained from the whole time window, the first and second half and from each quarter separately. Synchronised SOAEs recorded by time domain averaging show exactly the same frequencies in the TEOAE spectra of all analysed time window quarters compared to SOAEs recorded by frequency domain averaging. Moreover, synchronised SOAEs can influence the response shape of the TEOAE additionally. This is shown in long lasting responses exceeding the inter stimulus interval (ISI). In this case superimposing of consecutive single responses takes place and the shape of the averaged response depends on the exact ISI value used. These effects were investigated by varying the ISI in steps of 0.1 ms. The ISI variation has more pronounced effects on the response shape at short ISI values (50 ms) and low stimulus levels (10 dB HL). Thus the wave form changes are not accompanied by a change of the frequency content poor phase cancellation takes place. This is confirmed by simulated phase interaction. It is suggested that the phase of an already synchronised SOAE has significant effects on the re-synchronisation of the SOAE by the next click. Both influences should be considered in TEOAE recordings if synchronised SOAEs are present. PMID- 8226339 TI - Volume regulation in cochlear outer hair cells. AB - Many cells placed in a hypotonic medium initially swell and then rapidly undergo a regulatory volume decrease (RVD) to return towards original volume. Re-exposure to the isotonic solution results in the cells shrinking followed by a regulatory volume increase (RVI). Previous studies have shown that isolated outer hair cells (OHCs) placed in a hypotonic medium swell and maintain this shape until returned to the original medium. We re-examined this apparent lack of cell volume regulation in OHCs. OHCs were isolated from guinea pig cochleae, mechanically dissociated and dispersed, and placed in a Hank's balanced salt solution (HBS). In the cells studied, switching the perfusate to a hypotonic HBS (290-280 mmol/kg) for 15 min resulted in an immediate shortening of the OHCs (i.e., volume increase). In 26% of the cells, this increase was followed by a return to original length during the time the cell was perfused with the hypotonic medium, a RVD. Twelve percent of the cells demonstrating a RVD also displayed a RVI. Omitting collagenase and increasing Ca2+ concentration did not increase the percentage of cells displaying a RVD, while gadolinium (Gd3+, 10 microM) decreased the percentage to zero. This is the first report of isolated OHCs undergoing cell volume regulation. PMID- 8226340 TI - The vascular component of sodium salicylate ototoxicity in the guinea pig. AB - Drugs of the salicylate family (aspirin-like drugs) are reversibly ototoxic. Electrophysiologic and ultrastructural evidence suggests an impairment of the sensory hair cells of the cochlea following salicylate treatment. In addition, since these drugs can cause vasoconstriction, the ototoxicity of salicylates may also involve an impairment of the blood circulation in inner ear. However, a vascular hypothesis of salicylate toxicity has not received much attention. In the current study, we simultaneously measured cochlear blood flow (by laser Doppler flowmetry) and the sound-evoked potentials from the round window. Sodium salicylate caused a decrease in cochlear blood flow that appeared within 30 min following an intramuscular injection of a low dose of sodium salicylate (100 mg/kg). This sodium salicylate dose did not cause a change in auditory sensitivity. For higher doses (200 mg/kg and 300 mg/kg), both cochlear blood flow and auditory sensitivity were affected. The 300 mg/kg dose decreased blood flow by about 25% and elevated compound action potential thresholds by 10 to 25 dB for high frequencies (> or = 8 kHz). Further experiments showed that salicylate induced threshold shifts were significantly reduced for the mid-frequencies when cochlear blood flow is increased by the vasodilating drug hydralazine (negating the flow reduction caused by salicylate). These data indicate that in addition to the direct effect of systemically administered salicylate on neurosecretory function a decreased blood flow contributes to the ototoxicity of salicylates. PMID- 8226341 TI - Receptor-mediated release of inositol phosphates in the cochlear and vestibular sensory epithelia of the rat. AB - Various neurotransmitters, hormones and other modulators involved in intercellular communication exert their biological action at receptors coupled to phospholipase C (PLC). This enzyme catalyzes the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PtdInsP2) to inositol 1,4,5-trisphosphate (InsP3) and 1,2-diacylglycerol (DG) which act as second messengers. In the organ of Corti of the guinea pig, the InsP3 second messenger system is linked to muscarinic cholinergic and P2y purinergic receptors. However, nothing is known about the InsP3 second messenger system in the vestibule. In this study, the receptor-mediated release of inositol phosphates (InsPs) in the vestibular sensory epithelia was compared to that in the cochlear sensory epithelia of Fischer-344 rats. After preincubation of the isolated intact tissues with myo [3H]inositol, stimulation with the cholinergic agonist carbamylcholine or the P2 purinergic agonist ATP-gamma-S resulted in a concentration-dependent increase in the formation of [3H]InsPs in both epithelia. Similarly, the muscarinic cholinergic agonist muscarine enhanced InsPs release in both organs, while the nicotinic cholinergic agonist dimethylphenylpiperadinium (DMPP) was ineffective. The muscarinic cholinergic antagonist atropine completely suppressed the InsPs release induced by carbamylcholine, while the nicotinic cholinergic antagonist mecamylamine was ineffective. Potassium depolarization did not alter unstimulated or carbamylcholine-stimulated release of InsPs in either organ. In both tissues, the P2 purinergic agonist alpha,beta-methylene ATP also increased InsPs release, but the P1 purinergic agonist adenosine did not. These results extend our previous observations in the organ of Corti of the guinea pig to the rat and suggest a similar control of the InsP3 second messenger system in the vestibular sensory epithelia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226342 TI - Expression of NMDA-receptor mRNA in the rat cochlea. AB - While there is considerable evidence that an excitatory amino acid and excitatory amino acid receptors are involved in the synapse between inner hair cells and the auditory nerve, evidence for the specific involvement of the N-methyl-D-aspartate (NMDA) receptor is more ambiguous. With the cloning of the NMDA receptor, probes are now available that can determine in which neurons the receptor is being expressed. In situ hybridization histochemical techniques were therefore utilized to examine the expression of NMDA receptor messenger ribonucleic acid (mRNA) in the rat cochlea. Expression of NMDA receptor mRNA was seen in spiral ganglion cells. These results suggest that NMDA receptor is a component of excitatory amino acid synapses in the cochlea. PMID- 8226343 TI - The sensitivity to 3,3'-iminodipropionitrile differs for high- and midfrequency hearing loss in the developing rat. AB - 3,3'-Iminodipropionitrile (IDPN) has been demonstrated to produce a loss of hearing following both neonatal and adult exposures. Adult exposure induces a full spectrum hearing loss, whereas early postnatal exposure produces a high frequency loss only. The purpose of this work was to delineate the period of development during which the rat becomes sensitive to the full ototoxic effects of IDPN. Primiparous Long Evans rats or their offspring were exposed to either saline or 300 mg/kg IDPN for three consecutive days. Ages of exposure were as follows: gestational days 15-17 or postnatal days (PND) 1-3, 5-7, 15-17, 20-22, 25-27, 30-32, 40-42, or 70-72. All animals were tested as adults for auditory thresholds to 5- and 40-kHz tones using reflex modification audiometry. Results demonstrate that adult-like susceptibility to IDPN was not reached until approximately PND 30-32. Early exposures (PND 5-22) to IDPN will induce a highfrequency selective hearing loss, sparing the lower frequency. Prenatal or early neonatal (PND 1-3) IDPN exposure resulted in a high degree of mortality (> 70%). The long period of time between the susceptible period for the high frequency (PND 5-7) and the lower frequency (PND 30-32) does not correspond to the basal to apical ontogenic profile of any one physiological or anatomical process. These data suggest either a unique site of action for IDPN in the cochlea or the possibility of two different mechanisms, one operating at early postnatal ages and one at later ages. PMID- 8226344 TI - Thyroid hormone induces earlier onset of auditory function in neonatal rats. AB - The effect of thyroid hormone injection on the development of auditory function in neonatal rats was evaluated using auditory nerve-brainstem evoked responses (ABR). The hormone induced earlier onset of auditory function. In order to differentiate between conductive and sensorineural factors, both air-conducted (AC) and bone-conducted (BC) ABR responses were recorded. Neonatal rats were injected with thyroxine (T4), or with saline (control animals), from day of birth (post-natal day-PND-0), daily, until PND 9. AC- and BC-ABRs were recorded from PND 6 up to PND 20. It was found that both AC- and BC-ABR thresholds were lower in the T4-injected rats up to PND 15, after which no difference was found between the two groups. This indicated earlier maturity of both conductive (external and middle ears) and sensorineural (inner ear) factors and is probably due to the earlier appearance in the blood of higher T4 levels, following injection, than that occurring naturally during the neonatal period in these animals. PMID- 8226345 TI - Development of hearing in neonatal rats: air and bone conducted ABR thresholds. AB - While the human full-term neonate can hear at birth, in the rat the onset of auditory function as monitored by recording auditory nerve-brainstem evoked responses (ABR) has been reported to begin on post-natal day (PND) 12-14 and reaches adult thresholds at about 22 days. In order to determine the factors involved in this late onset and then rapid threshold improvement in rats, the ABR to both air conducted (AC) and bone-conducted (BC) auditory stimulation was determined in neonatal rats. ABR to maximal intensity BC stimuli (55 dB above adult rat ABR threshold--55 dB HL*) could be recorded from PND 7-8 while AC responses to 80 dB HL* stimuli, only from PND 11. The air-bone gap (a measure of conductive immaturities only) disappeared on PND 15. This shows that there are both conductive (external and middle ear--Air-bone gap) and sensori-neural (inner ear--BC threshold) immaturities in the neonatal rat; the conductive factors are resolved by PND 15 while the sensori-neural continue after that. With respect to conductive factors, it seems that the state of the ear canal is not important while the chief conductive factors involved probably include mesenchyme resorption and/or ossicular ossification. The chief sensori-neural factor may be the development of the endocochlear potential. It is likely that the human fetus in-utero undergoes similar stages of development. PMID- 8226346 TI - Acute effects of cocaine on cochlear function. AB - The effects of a single administration of cocaine on the cochlea was evaluated by measuring amplitude-intensity functions of the N1 response of the auditory nerve. Amplitude-intensity functions of the N1 response to tone-pips of 500 Hz, 1, 2, 4 and 8 kHz were obtained before and after intraperitoneal injection of either saline, 3 mg/kg or 25 mg/kg of cocaine. N1 amplitudes were decreased after the administration of cocaine and this reduction was found to be dose dependent. The influence of cocaine on cochlear blood flow (CBF) was examined in order to test the possibility that cocaine induced reductions in CBF underlie these electro physiological changes. Corresponding decreases in cochlear blood flow after cocaine exposure were observed. PMID- 8226347 TI - The growth of hearing loss in neonatal chicks exposed to intense pure tones. AB - One-day-old chicks were exposed to an intense pure tone (0.9 kHz, 120 dB SPL) and assigned to one of eight groups based on continuous exposure durations from 1 to 200 h. As each animal was removed from the exposure, it was anesthetized and an electrode was placed in the nucleus magnocellularis. Sound-evoked potentials were used to measure absolute thresholds and frequency selectivity. Thresholds were measured at 10 frequencies between 0.1 and 4.5 kHz while frequency selectivity was assessed by a simultaneous masking tuning-curve procedure at five probe-tone frequencies between 0.3 and 2.5 kHz. Threshold shift was greatest at 1.3 kHz and reached a maximum loss of approximately 57 dB between 48 and 200 h of exposure. The shape of the threshold-shift curve as a function of exposure duration (for frequencies between 0.9 and 2.5 KHz) suggested the presence of an early and late effect. The loss in tuning sharpness was evaluated by comparing the values of Q10 dB for control and exposed tuning curves and expressing the difference between them as a percent change. Probe-tone tuning curves above 0.9 kHz became less selective as exposure duration increased. A maximum decrease in tuning of about 54 percent was reached by 48 h and this remained constant to 200 h. The low frequency tuning curves (below 0.9 kHz) did not show any systematic loss in selectivity. The changes in sensitivity and selectivity are discussed in relation to the patterns of cochlear injury that occurred on the basilar papilla as exposure duration lengthened. PMID- 8226348 TI - Characterization of wave I of the electrically evoked auditory brainstem response in the guinea pig. AB - This paper examines the first component of the electrically evoked auditory brainstem response (EABR) of the guinea pig. Short (20 microseconds/phase) and long (4000 microseconds/phase) duration rectangular current pulses were applied through a bipolar intracochlear electrode in acute preparations. Short-duration pulses evoked a synchronized response relatively free of stimulus artifact; long pulses facilitated examination of the integrative capacities of nerve fibers at relatively low current levels. In deafened control subjects, wave I of the EABR consistently demonstrated two positive peaks having different latency-level and adaptation recovery functions. The early component (wave Ia) showed less decrement in latency with increasing stimulus level and recovered faster in a forward-masking paradigm. Non-monotonicities in the adaptation recovery curves were also observed, more consistently in the wave Ib data. It is proposed that wave Ia arises from stimulation of the axons proximal to the spiral ganglion while wave Ib is initiated at the peripheral dendritic processes. Implications for human cochlear implant research are discussed. PMID- 8226349 TI - Physiological overshoot and the compound action potential. AB - The sensitivity of the compound action potential (CAP) of the auditory nerve of the Mongolian gerbil (Meriones unguiculatus) to an added signal was studied as a function of the state of adaptation to a masker. The masker consisted of a train of tone bursts. A signal was added to one of the bursts and the threshold for detecting the signal was determined using a two-interval forced-choice technique. With a signal at the same frequency as the masker, there was little or no change in threshold with increasing adaptation to the masker, i.e., with the signal applied at masker onset or later on in the train. With a signal at a different frequency, considerably higher thresholds were obtained in the unadapted state than in the adapted state, a situation that appeared to resemble psychophysical overshoot phenomena. However, the interpretation of this result was complicated by the observation that the CAP response to a two-frequency stimulus is a sum of two slightly desynchronized waveforms arising from different cochlear regions. Synchronizing the two waveforms reduced the 'overshoot', but still resulted in higher thresholds in the unadapted state. The differences in threshold are accounted for by considering the changing signal-to-noise ratios at different states of adaptation. Possible relationships to psychophysically observed overshoot are discussed. PMID- 8226350 TI - Effects of spike discharge history on discharge probability and latency in frog basilar papilla units. AB - Gaumond et al. [(1982) J. Neurophysiol. 48, 856-873] showed in the cat that a multiplicative-intensity model can generally account quite well for reduction of the probability of an auditory-nerve spike by another spike preceding it by 4 to 25 ms, and that for smaller separations there is also an increased latency of the following spike. Bosch [(1990) D. Sc. Dissertation, Washington University, St. Louis, MO] made important improvements in experimental design and estimation techniques for studying these effects, and confirmed their presence in the gerbil. However, direct application of these methods to the frog does not yield reliable estimates. A clearer separation of discharge probability and latency effects in frog basilar papilla units is provided by the paired-click paradigm used in this study, which is applicable to low-spontaneous-rate units that generally respond to click stimuli with zero or one spike within a short interval following the click. The results confirm the existence in the frog of both spike probability and spike-latency effects that are qualitatively similar to those found in mammals, although the absolute refractory time is much longer in frog, and the relative refractory time usually shorter. The paired-click paradigm also reveals a stimulus-history effect at stimulus levels which are near threshold: when there is no response to the first click, responses to the second click occur with increased probability and reduced latency. PMID- 8226351 TI - Choline acetyltransferase in cochlear root neurons. AB - Cochlear root neurons are a distinct group of cells located in the auditory nerve root in small rodents. Their transmitter is still unknown. Some of our preparations showed immunoreactivity of somata of cochlear root neurons with both polyclonal and monoclonal antibodies against choline acetyltransferase (ChAT) which, despite their very weak histochemical reaction for acetylcholinesterase (AChE), suggested that cochlear root neurons might be cholinergic. To test this, we used a radiometric assay to measure ChAT activities of rat auditory nerve root samples containing cochlear root neurons and of adjacent samples not containing them. There was no significant difference between the low mean ChAT activities of these two groups of samples. Thus, cochlear root neurons are not likely to be cholinergic. PMID- 8226352 TI - Simultaneous masking in newborn chickens. AB - Simultaneous masking was used to investigate the development of frequency analysis in newborn animals. The ability of young chickens to detect a tone in the presence of a second tone was measured. In the first experiment data were collected with a 500-Hz masker presented at 15 or 25 dB above absolute threshold. Signals to be detected varied up to an octave from the masker (+/- 1/9, +/- 2/9, +/- 1/3, +/- 2/3, and -1 octave). Receiver operating characteristics (ROCs) were constructed from delays in ongoing peeps that occur when chicks detect the presence of the signal over the masker. Psychometric functions were fit by probit analysis using areas under these ROCs. Amounts of masking were estimated by the degree to which psychometric functions were shifted toward the right by the presence of the masker. There is no developmental change in these masking patterns, corroborating recent data from human infants. Masking was shown in a second experiment to be similar at a higher frequency (given intensities corrected for improving absolute thresholds). In conclusion, masking patterns derived from chicks' peeps are like those from adult humans. Frequency analysis, as measured by simultaneous masking, appears to be mature in chickens at hatching. PMID- 8226353 TI - Localization of calretinin mRNA in rat and guinea pig inner ear by in situ hybridization using radioactive and non-radioactive probes. AB - The localization of calretinin mRNA was studied in the rat and guinea pig inner ear by in situ hybridization, and compared to the distribution of the protein previously examined by immunocytochemistry. Radioactive and non-radioactive in situ hybridization (ISH) were performed using oligonucleotide probes labelled with 35S or digoxigenin. Radioactive ISH was more sensitive than non-radioactive ISH. In cochlear and vestibular ganglia, calretinin mRNA was localized in subpopulations of neurons with patterns of distribution similar to those shown by immunocytochemistry. By contrast, the observations in the sensory epithelia differed with the two techniques, ISH revealing less positive structures than immunocytochemistry. Rat inner hair cells and guinea pig inner hair cells, Hensen's cells and Deiters cells, which had been described strongly immunoreactive, appeared positive with radioactive but not with non-radioactive ISH. On the other hand, rat vestibular type II hair cells and guinea pig interdental cells of the spiral limbus which were faintly immunoreactive were not positive with both ISH techniques. PMID- 8226354 TI - Stimulation or blockade of the dorsal nucleus of the lateral lemniscus alters binaural and tonic inhibition in contralateral inferior colliculus neurons. AB - Recent studies have demonstrated that several specific types of acoustically evoked GABA-mediated inhibition occur in neurons of the central nucleus of inferior colliculus (ICc). The dorsal nucleus of the lateral lemniscus (DNLL) provides a major GABAergic projection to ICc. The present study examined the effects of electrical or chemical stimulation or reversible blockade within the DNLL on the discharge characteristics of ICc neurons in anesthetized rats. Microinjection of a local anesthetic (lidocaine) or a GABA-A agonist (THIP) via a cannula placed into DNLL reversibly blocked acoustically-evoked binaural inhibition and increased spontaneous firing in most contralateral ICc neurons. Trains of electrical pulses or microinjection of the excitant amino acid, kainate, into DNLL resulted in reduced acoustically-evoked firing, which was similar to binaural inhibition, in most contralateral ICc neurons examined. The effects of DNLL electrical stimulation were reversibly blocked by microinjection of THIP into the stimulation site, suggesting that the effect of the electrical stimulation is mediated by direct effects on cell bodies of DNLL neurons. These data support the idea that contralateral GABAergic input from the DNLL is inhibitory to ICc neurons. Thus, binaural inhibition and tonic inhibition in ICc neurons may be mediated, in part, by the GABAergic projection from the contralateral DNLL. PMID- 8226355 TI - Norgestomet and gonadotropin-releasing hormone enhance corpus luteum function and fertility of postpartum suckled beef cows. AB - In Exp. 1,101 suckled beef cows were administered two injections of prostaglandin F2 alpha (PGF2 alpha) 11 d apart. Cows in Group 1 received no additional treatment. Cows in Groups 2 and 3 received 250 micrograms of GnRH plus 47-h calf removal. Cows in Group 3 also received a norgestomet ear implant that was inserted 4 d after the first PGF2 alpha injection and left in situ for 8 d. The second PGF2 alpha injection was administered approximately 28 h before the time of norgestomet implant removal. The GnRH was administered approximately 30 h after the time of norgestomet implant removal (or 58 h after the second PGF2 alpha injection). Calf removal was the period from the time of implant removal to AI. All cows were artificially inseminated once 75 h after the second PGF2 alpha injection (47 h after implant removal). Reproductive status before the time of GnRH treatment (presynchronization) was determined by multiple sampling (2, 13, and 23 d before GnRH treatment) for blood progesterone concentrations (62% anestrous and 38% cyclic). The GnRH treatment increased (P < .01) the ovulation response and norgestomet reduced (P < .01) the incidence of short luteal phases in the presynchronization anestrous cows. Norgestomet treatment increased (P < .05) the timed breeding pregnancy rates for both presynchronization anestrous and cyclic cows. In Exp. 2, 174 suckled beef cows were administered Syncro-Mate B (which includes norgestomet). Cows in Group 1 received no additional treatment, whereas cows in Group 2 received 250 micrograms of GnRH (30 h after norgestomet implant removal).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226356 TI - Reproductive efficiency of range beef cows fed different quantities of ruminally undegradable protein before breeding. AB - Two studies, using 128 crossbred, multiparous beef cows grazing native winter and spring rangeland, were conducted to determine effects of protein supplementation on nutrient status and subsequent calf and reproductive performance. Postparturient cows (body condition score [BC] = 3.9) were fed 1.82 kg of one of two supplements (54% CP at 490 g of CP/d) per cow on alternate days. Supplements were formulated to contain 50% (245/490 g of CP) ruminally undegradable (UD) CP (RU50) or 25% (119/490 g of CP) UD CP (RU25) and were fed until breeding (BC = 4.2). Late-calving cows (Study 1) were individually fed supplement while grazing native range. Early-calving cows (Study 2) were group-fed supplement on alternate days while grazing native range and were daily fed approximately 5.4 kg of medium quality grass hay (10.5% CP). Late-calving cows had similar (P > .22) BW losses postpartum, whereas early-calving cows fed RU50 lost 39 kg less (P < .01) BW than did cows fed RU25. Cows that received RU25 returned to estrus sooner (P < .02), in the late-calving study, than cows fed RU50 (47 vs 56 d, respectively). The percentage of cows serviced in the first 21 d of breeding (96 and 100% for late- and early-calving studies, respectively) and pregnancy rates (88 and 86% for late and early-calving studies, respectively) did not differ regardless of postpartum protein supplement. When adequate CP was provided in the diet for optimum function of the rumen, addition of ruminally undegraded CP decreased weight loss of mature, postpartum beef cows grazing native range.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226357 TI - Norgestomet implants synchronize estrus and enhance fertility in beef heifers subsequent to a timed artificial insemination. AB - Three trials involving 128 heifers were conducted to determine whether norgestomet implants administered during the mid- and late luteal phases after breeding could be used to synchronize a second estrus in nonpregnant, inseminated heifers without adversely affecting pregnancy in pregnant heifers. All heifers were initially synchronized with Syncro-Mate B and artificially inseminated 47 h after implant removal. On d 9 (Trial 1) or d 12 (Trial 2) after the timed AI, the heifers were randomly assigned to treated or control groups. Treated heifers received two silicone implants containing 10.0 mg of norgestomet each (Trial 1) or one silicone implant containing 3.6 mg of norgestomet (Trial 2). Silicone implants were removed on d 21 after the initial AI. In Trial 1, the calving rate to the initial AI of the control heifers was 35 vs 55% for the norgestomet implanted heifers (P > .05). In Trial 2 the calving rate to the initial AI of the control heifers was 9 vs 45% in the treated heifers (P < .01). At the return estrus 52% of the control heifers returned to estrus within a 3-d period, whereas 93% of the norgestomet-treated heifers returned to estrus within a 3-d period (P < .01). Norgestomet treatment had no effect on serum progesterone concentrations of the pregnant heifers on d 21 after the initial AI. In Trial 3, both control and treated heifers were administered silicone implants containing 3.6 mg of norgestomet on d 12; additionally, the treated heifers received an injection containing 3.0 mg of norgestomet and 5.0 mg of estradiol valerate. Norgestomet implants were removed on d 21.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226358 TI - Evaluation of biological efficiency of free-grazing beef cows under semidesert conditions. AB - Effects of cow BW, hip height, and estimated genetic potentials (EBV) for weaning weight direct and milk on cow productivity, fecal OM output, OM intake, and efficiency (kilograms of calf BW/kilogram of OM intake by the cow) were evaluated with 44 free-grazing crossbred cows under semidesert conditions. Calf BW were measured during early, mid-, and late lactation. Data were collected in four periods: Period 1 = late spring (early lactation), Period 2 = late summer (mid lactation), Period 3 = mid-autumn (late lactation), and Period 4 = mid-winter (nonlactation). Calf BW increased linearly with cow BW (P < .01) in Periods 1, 2, and 3. Fecal OM output and OM intake increased with cow BW in Periods 2 (P < .01) and 4 (P < .01), and on average (P < .02). Overall efficiency decreased with increasing cow BW (P < .04). Taller cows excreted more fecal OM and had greater OM intake throughout the study (P < .02 to P < .11). Overall efficiency decreased with increasing cow hip height (P < .05). Weaning weight direct EBV of cows was related linearly to cow BW (P < .01 to P < .07) and to calf BW (P < .01 to P < .07). Calf weight in all periods increased linearly with milk EBV (P < .001). Overall, fecal OM output, OM intake, and efficiency were not affected by milk EBV. PMID- 8226359 TI - Crambe meal is equivalent to soybean meal for backgrounding and finishing beef steers. AB - Growth, feed efficiency, and carcass characteristics of 70 crossbred steers fed one of four diets were compared. The four diets differed in source of protein supplementation: 100% soybean meal (SB), 67% SB and 33% crambe meal (CM), 33% SB and 67% CM, and 100% CM. All supplements were fed in isonitrogenous amounts. Steers were fed backgrounding diets (12.9% CP) for 84 d and finishing diets (11.2% CP) for 96 d. Average initial weight was 303.4 kg. Backgrounding gains ranged from 1.38 to 1.41 kg/d (P = .92). Finishing gains ranged from 1.43 to 1.47 kg/d (P = .86). Range in entire-experiment gains was 1.41 to 1.46 kg/d (P = .85). Feed efficiencies were equal; entire-experiment efficiencies averaged .144 (P = .96). Growth and efficiency patterns were the same for all four treatments. No overall treatment differences were detected for the seven carcass variables (P = .26 to .96). Average fat depth, longissimus muscle area, yield grade, and dressing percentage were .95 cm, 84.0 cm2, 2.45, and 61%, respectively. At the protein percentage levels of these diets, CM substituted equally for SB for growth rate, feed efficiency, and carcass characteristics. PMID- 8226360 TI - Genetic parameters for growth traits of Australian beef cattle from a multibreed selection experiment. AB - Estimates of covariance components and genetic parameters were obtained for birth, weaning, 200-d, and 400-d weight for a herd of Polled Herefords and a herd of a multibreed synthetics, the so-called Wokalups. Data originated from an experiment in Western Australia selecting for preweaning growth rate. Analyses were carried out by REML fitting an animal model including genetic and permanent environmental maternal effects. Wokalups showed consistently more phenotypic variation, partly due to a scale effect, and higher direct and lower maternal heritabilities than Herefords. Maternal environmental effects were more than twice as important in Herefords than in Wokalups. Estimates of both genetic and environmental correlations among weaning and subsequent weights were essentially unity, identifying maternal effects found for postweaning weights as a "carry over" of those on weaning weight. PMID- 8226361 TI - Genetic parameter estimates of ultrasound-measured longissimus muscle area and 12th rib fat thickness in Brangus cattle. AB - Data from 2,101 Brangus calves born from 1986 to 1990 were analyzed with a REML procedure using a derivative-free algorithm in a mixed linear animal model to obtain variance component estimates of ultrasound-measured longissimus muscle area and fat thickness. Direct additive heritabilities (ha2) of .39 and .40 were obtained for age-constant weaning and yearling longissimus muscle area (WLMA and YLMA, respectively), with a genetic correlation (rg) of .66 between them. The rg of YLMA with birth weight (BWT), weaning weight (WWT), postweaning gain (PWG), yearling weight (YWT), frame score (FS), and scrotal circumference (SC) were .17, .29, .43, .38, .01, and .19, respectively. The ha2 of age-constant yearling 12th rib fat thickness (FAT) was .14, and cattle averaged .44 cm (SD = .19). Positive rg were obtained between FAT and WLMA (.19) and YLMA (.12). Negative rg of FAT with WWT, YWT, and SC were -.17, -.53, and -.33, respectively. Positive rg were obtained between FAT and BWT (.52), PWG (.44), and FS (.14). Maternal heritabilities (hm2) of WLMA, YLMA, and FAT were .01, .01, and .10, respectively. Weight-constant WLMA, YLMA, and FAT ha2 were .36, .39, and .11, respectively. Selection based on either age-constant YLMA or FAT could potentially result in 1.06 cm2 or .005 cm change per year, respectively, which would be slightly greater than change from selection based on weight-constant YLMA or FAT. Selection based on WLMA or YLMA should be effective, and changes in these traits, growth, and SC should be possible in tandem.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226362 TI - The effect of selection in sublines and crossing on genetic response and inbreeding. AB - More intense selection in a line gives greater genetic change but also gives a higher rate of inbreeding. Here, continuous selection in one line vs continuous selection in two or more sublines (for the same breeding objective and with the same total number of individuals tested) and crossing the sublines to reduce inbreeding, is studied by stochastic simulation using an infinitesimal model. On crossing, the inbreeding level in the cross, though zero, becomes equivalent to 1/n times that in the sublines, where n is the number of lines in the cross. In general, selecting in sublines and crossing gave less genetic response at the same level of inbreeding than selection in a single line. Use of the best subline generally gave less response than the whole line. It is concluded that selection response at the same level of inbreeding is greatest when all the test facilities and selection are concentrated in a single selection line. PMID- 8226363 TI - Enhanced lipolysis from broiler adipocytes pretreated with pancreatic polypeptide. AB - Broiler adipocytes in culture were used to determine whether prolonged preincubation with an antilipolytic hormone, pancreatic polypeptide, enhances lipolysis by inducing desensitization of lipolysis inhibition. Preincubation of broiler adipocytes with pancreatic polypeptide resulted in a dose-response and time-dependent enhancement (P < .05) of basal and glucagon-stimulated lipolysis. Lipolysis was enhanced at 4 and 24 h but not at .5 h of pretreatment. Acute inhibition of basal lipolysis was unaffected by long-term (24 h) exposure of adipocytes to a maximally effective dose (12 nM) of pancreatic polypeptide. Thus, desensitization of lipolysis inhibition cannot explain the enhanced lipolysis from pancreatic polypeptide-treated adipocytes. However, preincubation with 12 nM pancreatic polypeptide for 24 h reduced (P < .05) pancreatic polypeptide and somatostatin inhibition of lipolysis stimulated by glucagon. Moreover, basal lipolysis and submaximal lipolysis were enhanced to a similar extent (27 vs 29 nmol/h) but to a greater (P < .05) extent (27 vs 13 nmol/h) than maximal lipolysis when adipocytes were exposed to 12 nM pancreatic polypeptide for 24 h. These results suggest that the enhanced lipolysis induced by prolonged exposure of adipocytes to pancreatic polypeptide resulted from increased activity of hormone-sensitive lipase and activators of this enzyme and not from attenuation of lipolysis inhibition. PMID- 8226364 TI - Effect of selection for lean tissue growth on body composition and physiological state of the pig at birth. AB - The effects of selection for lean tissue growth on the metabolic and physiological state (i.e., level of maturity) of the pig at birth have been examined on newborns from three breeds that markedly differ with respect to birth weight and postnatal muscle growth potential: a primitive Chinese breed (Meishan, MS), a European breed (Large White, LW), and a composite line (CL) highly selected for high rate of gain. Within each breed, 40 pigs from eight litters were used for whole carcass and tissue sampling, blood sampling, and for a fat tolerance test at 2 h of age. The CL pigs were heavier (P < .001) than the LW and MS pigs at birth but exhibited lower percentages of carcass protein, fat, mobilizable fat, and ash than the MS pigs (P < .05). In addition, MS pigs had larger adipose tissue adipocytes than pigs from the two other breeds (P < .001). Despite their 31% higher RNA capacity in longissimus muscle (higher RNA:protein ratio, P < .05), CL pigs exhibited a lower percentage of muscle protein (P < .05) than did MS pigs. Relative liver weight was higher for LW than for CL pigs (P < .05), which had the lowest percentage of liver phospholipids (P < .01). The CL pigs exhibited lower hematocrit (P < .01), glucose (P < .01), albumin (P < .01), cortisol (P < .01), and thyroxine (P < .05) levels than the MS pigs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226365 TI - Use of chuck muscles and their acceptability in restructured beef/surimi steaks. AB - Ten major muscles along with any unidentifiable lean, were carefully excised from 16 Choice square-cut chucks Yield Grade 2, and placed according to previously determined tenderness rankings, into one of three muscle groups. Group 1 was composed of the most tender muscles, and contained the infraspinatus, longissimus, and triceps brachii. Group 2 contained intermediate tenderness muscles and was composed of the serratus ventralis, deep pectoral, and complexus. Group 3 contained the least tender muscles and was composed of the biceps brachii, supraspinatus, rhomboideus, trapezius, deltoids, and neck muscles. Each group was restructured into beef/surimi steaks and was evaluated. Total muscle yield before trimming accounted for 66.2% of the chuck. Careful fat trimming, desinewing, and internal seam cutting on individual muscles resulted in 34.7% lean available for the restructuring of steaks. The triceps brachii, longissimus, supraspinatus, and infraspinatus required the least trimming and were easiest to excise. These muscles made up 49% of the trimmed meat and 13.7% of the total chuck. Steaks were evaluated by a consumer sensory panel for tenderness, flavor, overall preference, and intent to purchase. There were no differences detected by consumers among the muscle groups for the sensory traits studied. Tenderness and flavor were rated equal to intact steaks for all muscle groups studied. The consumer sensory panel indicated that Groups 1 and 2 would be purchased twice a month and Group 3 once a month. PMID- 8226366 TI - The effect of trenbolone acetate on performance, plasma lipids, and carcass characteristics of growing ram and ewe lambs. AB - The purpose of the study was to determine the effect of trenbolone acetate (TBA) on performance, plasma lipid metabolites, and carcass characteristics of growing ram and ewe lambs in two separate experiments. Treatments consisted of 1) a control, no implants, and 2) a TBA implant (60 mg of Finaplix; Hoechst-Roussel Agri-Vet). Twelve Rambouillet cross ram lambs (average 19.9 kg BW; Exp. 1) and 18 Suffolk-Hampshire ewe lambs (average 37.2 kg BW; Exp. 2) were given ad libitum access to a 77% forage/23% concentrate diet (15.5% CP and 2.14 Mcal of ME/kg). On d 0, six rams and nine ewes received TBA as an ear implant and six rams and nine ewes were designated as controls. Blood samples were collected via jugular venipuncture on d 0, 28, 56, and 84 and plasma was separated and analyzed for lipid metabolites. Rams (30.5 kg BW) and ewes (50.0 kg BW) were slaughtered after 93 and 91 d, respectively. Average daily gain was not affected by TBA in rams (117 g/d) or ewes (159 g/d). Nutrient intake restricted normal BW gains in the rams. Plasma concentrations of total cholesterol, high-density lipoprotein cholesterol, triglycerides, and nonesterified fatty acids were not affected by TBA in either rams or ewes. Trenbolone acetate had no effect on subcutaneous fat measurements in either rams or ewes. Lamb chops from rams implanted with TBA were less (P < .04) tender (5.58 vs 4.26 kg/1.27-cm core) than lamb chops from control rams. Additional studies examining growth and carcass composition are needed in ruminants implanted with anabolic steroids and fed high-forage diets. PMID- 8226367 TI - Developing a fiber-optic probe to combine subcutaneous fat depth and meat quality measurements. AB - This feasibility study demonstrates that the boundary between porcine subcutaneous fat and longissimus muscle can be detected with a probe using optical fibers to illuminate and detect anatomical boundaries. A relatively large optical window (approximately 3 mm2) seemed to give the best resolution of fat to muscle boundaries, whereas a smaller window gave a better resolution of marbling. Scattered, obliquely sectioned optical fibers gave reflectance spectra of fat and muscle that were relatively flat and parallel (fat > muscle). Thus, white light may be used with optical fibers to detect fat to muscle boundaries, although the greatest monochromatic separation was at 820 nm. In comparing measurements made as the probe penetrated the meat with those made as the probe was withdrawn, structures appeared to be deeper on the way in than on the way out, because of compression of the meat. Multichannel operation allowed tissue transmittance to be measured as well as reflectance and channels to be averaged to improve performance. These observations will be useful in developing new apparatus to measure fat depth and meat quality in one operation using optical fibers. PMID- 8226368 TI - Effects of porcine somatotropin and dietary phosphorus on growth performance and bone properties of gilts. AB - One hundred eight gilts (initial weight = 58.5 kg) were used to determine the effects of porcine somatotropin (pST) and dietary P on growth performance and bone mechanical properties and mineralization during the finishing phase (58 to 105 kg) and a 35-d postfinishing phase. Gilts were injected daily with placebo (control) or 4 mg of pST and fed diets containing .4, .8, or 1.2% P in a 2 x 3 factorial arrangement. From 58 to 105 kg, administration of pST increased (P < .01) ADG and G/F and decreased (P < .01) ADFI. When mean weight of the gilts in a pen reached 105 kg, half the gilts were slaughtered and first rib, femur, and third and fourth metacarpals were collected for determination of mechanical properties and bone ash. A pST x P interaction was observed (P < .05) for rib bending moment and modulus of elasticity; maximum rib bending moment was attained by control gilts at .8% P and rib modulus of elasticity values remained constant across P levels, whereas rib bending moment and modulus of elasticity increased as dietary P increased from .4 to 1.2% in pST-treated gilts. Administration of pST decreased (P < .05) stress of the rib, femur, and metacarpals compared with control gilts. Increasing dietary P resulted in a linear (P < .10) increase in bending moment, stress, and ash content for rib, femur, and metacarpal bones. The remaining 54 gilts were individually fed 1.8 kg/d of a common diet for 35 d postfinishing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226369 TI - The interrelationship of porcine somatotropin administration and dietary phosphorus on growth performance and bone properties in developing gilts. AB - Seventy-two gilts (initial weight = 57.9 kg) were used to determine the interrelationship of porcine somatotropin (pST) administration and dietary P on growth performance of finishing gilts (58 to 106 kg) and the effect on bone mechanical properties and mineralization for a 35-d postfinishing phase after withdrawal of pST administration. Gilts were injected daily with placebo (control) or 4 mg of pST and fed .4, .6, or .8% P in the finishing phase. Administration of pST increased ADG and G/F (P < .01) and decreased ADFI (P < .01) during the finishing phase. Increasing dietary P resulted in increased (quadratic, P < .04) ADG from d 0 to 28 of the finishing phase; however, dietary P had no effect (P > .18) on ADG, G/F, or ADFI for the overall finishing phase. When each block weight averaged 106 kg, half the gilts were slaughtered and the first rib, femur, and third and fourth metacarpals were collected. Stress, modulus of elasticity, and ash content of rib, femur, and metacarpals were reduced (P < .06) and femur wall thickness was increased (P < .01) in pST-treated gilts. Increasing dietary P increased (linear, P < .05) bending moment, stress, and ash content for all bones collected, with the exception of metacarpal stress, which was not affected (P > .22). The remaining 36 gilts were individually fed 1.8 kg/d of a common diet to assure a P intake of 22.8 g/d for the 35-d postfinishing phase. Gilts that received higher levels of dietary P during the finishing phase had increased (linear, P < .06) bending moment and ash content for the rib and femur; rib stress, and femur wall thickness were also increased after the postfinishing phase. From d 0 to 28 of the finishing phase, pST-treated gilts required a diet with > .4% P (10.3 g/d P) to maximize growth performance. However, a diet with .4% P (12.44 and 10.66 g/d P, control and pST-treated, respectively) was adequate for growth performance during the overall finishing phase (56 to 106 kg).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8226370 TI - Dietary clofibric acid increases intestinal fatty acid binding protein activity and apparent lipid digestibility in weanling swine. AB - The effects of supplementation of dietary clofibric acid (.5% wt/wt) on fatty acid binding protein (FABP) activity, apparent lipid digestibility, and serum cholesterol concentrations were evaluated in weanling pigs. Twenty-four barrows were allotted by weight and litter to a randomized complete block design with two treatments (basal vs clofibric acid) in six replicates. Nutrient digestibility measurements were made for a 2-wk period, after which the pigs were killed and tissues were collected. No differences in BW, liver, proximal small intestine, distal small intestine, and proximal and distal intestinal mucosa weights were observed. Apparent lipid digestibility was greater (P < .05) for the overall 2-wk period in clofibric acid-supplemented pigs (81.5 vs 76.6%). This paralleled the increased FABP activity in the distal small intestine (P < .001) of clofibric acid-supplemented pigs. Proximal intestine and liver FABP activities were unaffected by dietary treatment. Serum cholesterol concentrations were markedly lowered by clofibric acid supplementation. During wk 1, pigs fed the basal diet had twofold greater (P < .01) serum cholesterol concentrations, whereas during wk 2, basal-fed pigs had fourfold greater (P < .01) serum cholesterol concentrations (81.5 vs 18.3 mg/dL). These results suggest that elevated intestinal FABP activities may augment fatty acid absorption from the gastrointestinal tract. PMID- 8226371 TI - Effect of concentration of mixed-function oxidase on concentration of estrogen, rate of egg lay, eggshell thickness, and plasma calcium in laying hens. AB - The objective of this study was to investigate the effect of induction or depression of mixed-function oxidases (MFO) by xenobiotics on the peripheral concentration of estrogens, plasma Ca, rate of egg lay, and eggshell thickness in laying hens. In Exp. 1, 100 mg of phenobarbital (PB) administered orally each day for 3 or 7 d reduced concentrations of estrone and estradiol (E2) in serum. In Exp. 2, 25, 50, or 100 mg of PB was administered orally to laying hens for 3 or 7 d. Treatment with 100 mg of PB for 3 d or with 25, 50, or 100 mg for 7 d significantly increased liver:body weight ratios. Microsomal protein increased after 7 d of 50 or 100 mg of PB administration. Feeding PB decreased concentration of E2 and total plasma Ca in a dose- and period-dependent manner. Concentration of E2 was reduced to 10% of control, whereas hepatic cytochrome P 450 increased significantly with dose. The correlation between concentration of E2 and P-450 was negative and significant. Total Ca in the plasma was highly, positively correlated with concentration of E2. Eggshells were thinner from hens treated with 100 mg of PB for both 3 or 7 d than those from control hens. Rate of egg lay was reduced by 100 mg of PB for 7 d. In Exp. 3, .5 mL of CCl4 given orally for 1 d decreased P-450 and increased E2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226372 TI - Pulsatile infusion of gonadotropin-releasing hormone initiates luteal activity in nutritionally anestrous beef cows. AB - Thirty-five nonlactating Hereford and Hereford x Angus cows weighing 469 +/- 8 kg, with a body condition score (BCS) of 5.5 +/- .6 and exhibiting normal estrous cycles, were fed to lose 1% of BW per week (n = 27) or to maintain BW (n = 8). Blood samples were collected every 7 d to monitor luteal activity and BW and BCS were recorded every 2 wk. When concentrations of progesterone in plasma were < 1 ng/mL for three consecutive weeks, anestrous cows (BCS, 3.1 +/- .7; BW loss, 24 +/- 5%) were assigned to pulses (i.v.) of saline (2 mL every hour), GnRH-4 (2 micrograms of GnRH every 4 h), or GnRH-1 (2 micrograms of GnRH every hour). Cows were confined to stalls for 4 d and two jugular cannulas were inserted 2 d before infusion of GnRH. Infusions began on d 0 and continued through d 14. Progesterone was quantified in daily blood samples and LH was quantified in samples collected every 10 min for 4 h on d-1, 0, 2, 4, 6, 8, 10, 12, 14, and 15. On d 16, cows were returned to pasture and blood samples were collected every 2nd d through d 30. Pulsatile infusion of GnRH increased (P < .05) the amplitude of LH pulses but did not affect pulse frequency. During infusion of GnRH every hour or every 4th h, concentrations of LH in serum were increased compared with those in saline treated cows (P < .001), and cows receiving GnRH hourly had greater (P < .005) LH than cows given GnRH every 4 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226373 TI - Effects of somatotropin treatment on growth performance, carcass traits, and the endocrine system in finishing beef heifers. AB - The objectives of this study were to investigate the effects of exogenous somatotropin on growth performance and carcass traits and on concentrations of somatotropin (ST), IGF-I, insulin, glucagon, free fatty acids (FFA), and glucose in plasma of finishing Simmental beef heifers. Three treatment groups, which included 12 heifers each, received a subcutaneous injection of a slowly released recombinant bovine somatotropin (rbST) at two doses (320 and 640 mg, respectively) or a placebo at 2-wk intervals. Treatments began at an average BW of 286 kg and continued until the animals were slaughtered at approximately 520 kg. Heifers had ad libitum access to corn silage and received 1.8 kg/d of concentrate. A before-feeding blood sample was collected from the jugular vein immediately before and 1 wk after each injection during treatment. Treatment tended to increase ADG and to reduce energy consumption per kilogram of gain, whereas feed and energy intake were not altered. Dressing percentage and conformation did not seem to be affected by treatment. Treatment led to a dose dependent reduction of fat tissue in carcass as well as in kidney fat and to a simultaneous increase of lean tissue (P < .05). Furthermore, treatment caused moderate increases of ST in both treated groups, whereas IGF-I concentrations increased markedly soon after the beginning of the treatment. Insulin and FFA concentrations were elevated in treated groups compared with controls. Glucagon concentrations were not altered by treatment. Effects on growth performance and carcass characteristics are discussed in view of metabolic and endocrine changes. PMID- 8226374 TI - Reproductive, endocrine, and organ weight differences of swine selected for high or low serum cholesterol. AB - Three generations of selection for 56-d blood cholesterol concentrations were used to establish low and high cholesterol lines of pigs in which cholesterol concentrations differed by 39% in the last generation. Litter size (number of fully formed pigs born per litter) diverged with each successive generation of selection and, at the third generation, litter size differed between the low and high line by two pigs (high line = 8.5 +/- .6; low line = 10.5 +/- .5; P < .05). A random selection of offspring (gilts, n = 109; boars, n = 46; barrows, n = 94) from the third generation and a control line maintained throughout the experiment were monitored for hormonal and anatomical relationships that might provide insight into the mechanisms that altered fecundity. Ovulation rate (number of corpora lutea) as determined on d 60 of pregnancy was increased in the low (n = 29) compared with the high cholesterol line (n = 38; 11.8 +/- .3 vs 9.8 +/- .3, respectively; P < .05), and litter size continued to be increased in the low cholesterol line (P < .05). No differences were noted between lines in kidney, ovarian, or total corpora lutea weight. Empty uterine weight and adrenal weight were increased in the high cholesterol line (P < .05) in randomly selected offspring of the third generation, and liver weight increased in the low line (P < .05). Associated with increased concentrations of cholesterol were increased serum concentrations of progesterone (gilts) and testosterone (boars) in the high cholesterol line (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226375 TI - Nycterohemeral eating and ruminating patterns in heifers fed grass or corn silage: analysis by finite Fourier transform. AB - Average daily and within-day nycterohemeral patterns of eating and ruminating behavior were determined in six Holstein-Friesian heifers (average BW = 427 kg) given ad libitum access to either corn or grass silage in a two-period crossover design. Rhythm components (number of cycles/24 h) were characterized by finite Fourier transform of the 24-h mastication activities as measured during 4 d by continuous jaw movement recordings. Average daily voluntary intake of corn silage was 8.2% greater (P = .05) than that for grass silage and was associated (P < .05) with fewer meals and shorter daily, unitary eating and ruminating times, and smaller number of rumination boli. Analysis of variance of the daily mean of hourly activities and Rhythm Components 1 to 12 indicated effects of (P < .05) silage type (S), animal (A), period (P), and a significant interaction (S x A x P) for each mastication activity. The finite Fourier transform was reparameterized to express the amplitude (as periodograms) and phase of each rhythm component. Rhythm Components 1, 3, and 4 contributed primarily to explaining the total dispersion of the 24-h series of time spent eating and ruminating, for both silage types and individual heifers. Relative importance of Rhythm Component 1 of time spent eating, indicative of a main circadian pattern, was related positively to pedigree value for milk production (P = .01) and negatively to milk protein concentration (P = .09).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226376 TI - Effect of copper level and source (copper lysine vs copper sulfate) on copper status, performance, and immune response in growing steers fed diets with or without supplemental molybdenum and sulfur. AB - One hundred twenty-six crossbred steers (218 kg initial BW) were used to determine the availability of Cu from copper lysine (CuLys) relative to CuSO4. Steers were assigned to pens (four replicates per treatment) based on BW and initial plasma Cu concentration and fed a corn silage-based diet supplemented with 0 or 5 ppm of Cu from either CuSO4 or CuLys. Half of the steers in each treatment were supplemented with 5 ppm of Mo and .2% S. Molybdenum and S supplementation increased (P < .10) growth rate during the first 21 d. Steers receiving CuSO4 gained more during the first 21 d than did control steers (P < .10) and steers receiving CuLys (P < .01). Growth, feed efficiency, and feed intake were not affected over the entire 98-d trial. Molybdenum and S supplementation decreased (P < .05) plasma Cu concentrations. Plasma Cu concentration was not affected by Cu source. Humoral immune response to ovalbumin was measured on d 7 and 77. Dietary treatment did not affect antibody production at either time. Cell-mediated immunity was measured in vivo on d 7 and 77 using phytohemagglutinin. In vivo cell reactivity was not affected by treatment on d 7 but was reduced (P < .10) by Mo and S supplementation on d 77. In vitro cell reactivity was measured on d 98 using a lymphocyte blastogenesis assay. Unstimulated lymphocytes from steers supplemented with Mo and S had lower (P < .10) uptakes of [3H]thymidine. There were no differences among treatments when lymphocytes were stimulated with pokeweed mitogen or phytohemagglutinin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226377 TI - Technical note: comparison of techniques for measurement of oxygen uptake by cattle. AB - The objective of this study was to compare techniques for measuring whole-body O2 uptake of steers. Oxygen uptake provides a basis for evaluating metabolic energy expenditure. We measured O2 uptake as the product of the difference in O2 contents of incoming and exhaust air times air flow through a respiration calorimeter (head box). Additionally, we used catheters and thermistors placed into the blood vessels near the heart to measure cardiac output and whole-body O2 uptake. The cardiac output measurement system used thermodilution to measure the blood flow rate through the pulmonary artery. Catheters placed in the pulmonary artery and in the aorta provided respective samples of oxygen-depleted and oxygen enriched blood. The product of blood flow rate times blood oxygenation by the lungs provided the basis for calculating total O2 uptake. Four sets of simultaneous measurements with the two techniques were made on eight steers weighing from 272 to 528 kg. Oxygen uptake was not different (P > .20) for the two techniques. Overall (mean +/- SEM) O2 uptake measured by cardiac output (117.2 +/- 7.0 L/h) was 97 +/- 3% of O2 uptake measured by respiration calorimetry. PMID- 8226378 TI - Influence of forage species and diet particle size on the passage of digesta and nylon particles from the reticulorumen of steers. AB - To assess factors that influence the passage of digesta from the reticulorumen, ruminally fistulated steers (387 kg) were fed to appetite in a 4 x 4 Latin square design either orchardgrass (OG) (Dactylis glomerata L.) or switchgrass (SG) (Panicum virgatum L.) hays. The two hays were fed in either the long form or ground to pass a 2-cm screen. Steers fed OG diets had greater DMI (P < .05) than steers fed SG diets (11.7 vs 10.4 kg/d respectively). Particle size of ruminal digesta and feces determined by wet sieving was greater (P < .05) for the OG and long hay diets than for the SG and ground diets. Particle size of digesta collected from the anterior dorsal sac and anterior ventral sac of the rumen and from the reticulum did not vary (P > .25) with site. Conversely, ruminal concentrations of 1-mm nylon particles tended (P = .11) to be greater in the lower strata of the reticulorumen at 12 and 24 h after dosing. Neither a site effect nor a site x time-after-dosing interaction (P > .75) was observed for the 3- or 5-mm nylon particles, suggesting that a sorting process independent of specific gravity occurs in the rumen for smaller particles (before the reticulomasal orifice). Passage rate of the 1-mm nylon particles from the reticulorumen was greater (P < .01) for animals fed the SG than for those fed the OG diets, whereas for the 5-mm particles, passage rate was greater for the OG diets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226379 TI - Effects of an Aspergillus oryzae fermentation extract and other factors on lactate utilization by the ruminal bacterium Megasphaera elsdenii. AB - The objective of this study was to determine the effects of an Aspergillus oryzae fermentation extract (Amaferm) as well as other factors on lactate utilization by the ruminal bacterium Megasphaera elsdenii B159. Addition of Amaferm or a filter sterilized Amaferm filtrate stimulated L-lactate uptake by both M. elsdenii and the ruminal selenomonad strain H18. Growth of M. elsdenii in medium that contained DL-lactate (2 g/L), Trypticase, and yeast extract was only slightly stimulated by the addition of 5% (vol/vol) Amaferm filtrate after 24 h. However, growth of M. elsdenii in a similar medium lacking Trypticase and yeast extract was increased over twofold by the addition of either 2 or 5% (vol/vol) Amaferm filtrate. These results suggest that Amaferm provides growth factors (i.e., amino acids, B vitamins) to support growth of M. elsdenii on lactate. There was no inhibition of L-lactate uptake when lactate-grown cells of M. elsdenii were incubated with excess (10 mM) glucose, sucrose, or maltose. In addition, when cells were grown on glucose, sucrose, or maltose rather than lactate there was little difference in L-lactate uptake, suggesting that L-lactate transport in M. elsdenii is not subject to catabolite repression by these soluble sugars. Both K+ and Na+ had little effect on L-lactate uptake. Uptake was unaffected at extracellular pH values between 6.0 and 8.0, whereas pH values of 5.0 and 4.0 increased uptake. In addition, L-lactate uptake was inhibited between 34 and 61% by protonophores. These results suggest that protons may be involved in the uptake of L-lactate by M. elsdenii B159. PMID- 8226380 TI - Metabolizable protein and amino acid requirements of growing cattle. AB - Metabolizable protein and amino acid requirements for growing cattle were estimated using data from 11 research trials. A total of 543 steers were individually fed a high-roughage diet supplemented with protein at several levels above a urea supplement control. The mean weight for all animals was 253 kg, with a range in mean initial to final weights of 200 to 316 kg, respectively. Daily gain ranged from -.04 to .89 kg. Metabolizable protein for each treatment group was calculated at the point at which the protein requirement was met. The sum of dietary escape protein (basal and supplemental) and calculated microbial protein represented metabolizable protein supplied per test protein source analyzed in each trial. Daily gain was regressed against calculated metabolizable protein flow using weighted regression analysis (r2 = .69, n = 45) to determine the metabolizable protein requirements for maintenance (3.8 x BW.75 g/d, where BW is expressed in kilograms) and growth (305 g/kg of live weight gain). Calculated metabolizable amino acid requirements as a percentage of metabolizable protein for a 253-kg animal gaining .49 kg/d were as follows: methionine, 3.0%; total sulfur amino acids, 5.8%; lysine, 8.0%; tryptophan, 1.0%; threonine, 5.2%; valine, 5.7%; isoleucine, 5.6%; leucine, 6.9%; phenylalanine, 3.9%; and histidine, 1.6%. The proposed requirements were based on live animal gain and intake of metabolizable protein and should represent the needs of the growing beef animal. PMID- 8226381 TI - Net transfer of urea and ammonia across the ruminal wall of sheep. AB - Texel wethers (68 +/- 2.5 kg BW) fitted with catheters in the ruminal veins and a mesenteric artery, blood flow probes on ruminal arteries, and a ruminal cannula were fed 500 g of orchardgrass hay every 12 h. During the last third of the feeding cycle, intraruminal injections were performed to evaluate the effect of urease activity, osmolality, and concentrations of NH3, butyrate, and CO2 in the rumen on urea and NH3 fluxes across the rumen wall. At pH 6.7, NH3 absorption increased with NH3 and butyrate concentrations in the rumen, and to a lesser extent with CO2 concentration. The increase in ruminal blood flow associated with CO2 and butyrate increase was always greater than the increase in NH3 absorption. Increasing ruminal osmolality slightly decreased NH3 absorption. Ruminal NH3 concentration and ruminal blood flow seemed to be the main determinant of NH3 absorption. Decreasing urease activity in the rumen decreased urea net transfer. The net transfer of urea to the rumen was stimulated by CO2. High concentrations of NH3 (330 mg of N/L) and butyrate (25 mM) in the rumen decreased urea net uptake, whereas osmolality (up to 420 mOsmol/L) did not affect it. Modifications in ruminal blood flow or water net movement across the ruminal wall did not seem to account for the effect of CO2, NH3, and butyrate on urea net uptake. PMID- 8226382 TI - Effect of diet and folic acid addition on digestibility and ruminal fermentation in growing steers. AB - Eight ruminally fistulated steers (352 +/- 27 kg BW) were used in a replicated 4 x 4 Latin square design. Treatments were 1) 70% rolled barley +30% timothy hay (HC), 2) HC+folic acid (2 mg/kg BW; HC+F), 3) 30% rolled barley + 70% timothy hay (HF), and 4) HF+F. Total tract apparent digestibility of DM and CP were greater (P < or = .05) in steers fed the HC diet, whereas that of ADF and NDF was greater (P < .001) in steers fed the HF diet. Supplemental dietary folic acid had no effect on either of the forementioned variables, regardless of whether the animal received the HC or the HF diet. To compare ruminal DM disappearance, nylon bags containing either hay or barley were incubated in the rumen of each steer for 4, 8, 12, 24, and 48 h. Ruminal DM disappearance of hay and barley was affected (P < .05) by the type of diet fed but not by the addition of folic acid to either diet. Ruminal fluid was sampled on three consecutive days, at the following postprandial hours: 1, 2, 4, 8, 12, and 23. Differences in ruminal pH and individual VFA among treatments were attributable to the type of diet fed. Changes in ruminal VFA concentrations at various times after feeding were altered with folic acid addition; however, these modifications were not sufficient to affect in a significant way either ruminal or total tract digestibility. PMID- 8226383 TI - In vitro ouabain-sensitive respiration and protein synthesis in ruminal epithelial papillae of Hereford steers fed either alfalfa or bromegrass hay once daily. AB - Seven Hereford steers (457 +/- 8.5 kg BW) with ruminal cannulas were fed either long or short chopped bromegrass (B) or alfalfa (A) hay at 90% of a 3-h ad libitum intake once daily between 0900 and 1200 in a 4 x 4 complete and 3 x 4 incomplete Latin square design. After evacuation of ruminal contents, papillae from the ventral sac of the rumen were excised at 0900, 1200, and 2100 and placed in oxygenated media. Oxygen consumption was determined polarographically, after which either 10(-4) M ouabain or 10(-4) M cycloheximide was added for determination of ouabain-sensitive (OSO2) or cycloheximide-sensitive (CSO2) O2 consumption, respectively. Additionally, protein synthesis was measured by uptake of [3H]phenylalanine. Twenty-four-hour patterns of ruminal fluid pH, osmolality, ammonia N, and VFA were also determined. Steers fed A exhibited a rapid rise in total O2, OSO2, and CSO2 immediately after consumption of a meal; the respiration patterns of ruminal epithelial papillae from animals fed B lagged behind those observed for animals fed A. Patterns of O2 consumption for both diets paralleled those observed for ruminal concentration of products of fermentation; those fed A had a larger magnitude of change in both O2 consumption and fermentation products. Ruminal epithelial O2 consumption seems to be determined by substrate availability and products of fermentation, and Na+,K+ATPase and protein synthetic activity each account for one-fifth of ruminal papillae O2 consumption. Fractional rates of protein synthesis were unaffected by type of forage consumed. PMID- 8226384 TI - Effects of pregnancy on digesta kinetics and ruminal fermentation in beef cows. AB - Four pregnant and four nonpregnant, ruminally cannulated beef cows were used to evaluate the effects of the last trimester of pregnancy on digesta kinetics and ruminal fermentation. Before breeding, cows were allotted randomly either to pregnant (PR) or to nonpregnant (NP) groups; PR cows were bred at the first estrus after synchronization. All cows were fed long-stem fescue hay at 15 g of DM/kg of BW based on BW 120 d before parturition of pregnant cows (average BW of 642 kg). Collection periods, each lasting 9 d, began at 96, 68, 41, and 10 +/- 1.3 d before parturition. Particulate passage rate was greater (P < .05) and retention times and gastrointestinal fill were less (P < .05) for PR than for NP cows. Apparent total tract DM digestion, rate and extent (96-h in situ incubation) of NDF disappearance, and ruminal fluid kinetics were not affected (P > .10) by pregnancy. A physiological state x sampling day interaction was noted (P < .05) for ruminal NH3 N, total VFA concentrations, and molar proportions of butyrate. Ruminal pH and individual VFA proportions, however, did not differ (P > .10) between PR and NP cows, except for valerate, which was less (P < .05) for PR than NP cows. Results suggest that in late pregnancy with restricted feeding, passage rate of particulates increases without substantial changes in fermentation or extent of digestion. PMID- 8226385 TI - A review of behavioral factors involved in the development and continued performance of stereotypic behaviors in pigs. AB - Environmentally induced stereotypies, commonly observed in farm and zoo animals, are behaviors that are relatively invariant, that are regularly repeated, and that serve no obvious function. However, there is as yet no accepted means of discriminating between normal and abnormal behavior, and the assumption that stereotypies are abnormal may mask the fact that they arise in part through processes that "normally" control behavior. There is growing evidence that stereotypies in sows and broiler breeders are strongly related to feeding motivation. For example, sows only develop oral stereotypies if their feed intake is restricted, and operant conditioning experiments have shown commercial levels of feed restriction to give rise to high levels of feeding motivation. Stereotypies in animals whose feed intake is restricted largely occur in the postprandial period, and ingestion of food has specifically been shown to elicit stereotypies in sows. These observations suggest that positive feedback from feeding produces a short-term increase in feeding motivation that at the end of the meal is directed toward available, alternative stimuli such as chains, the choice of stimuli reflecting the sensory feedback from the activity. Drinking behavior may also become an expression of feeding behavior after metabolic water requirements are met. In addition to these processes specific to feeding motivation, it seems likely that nonspecific processes, which operate more generally across motivational systems, contribute to the persistence of the behavior. Behavioral arousal may facilitate performance of active behaviors, and sensitization of the underlying neural elements may lead to the behavior being more easily elicited and maintained. A crucial factor in the sensitization process would seem to be the channeling of complex behavior by the environment into a few and very often repeated sequences of behavior. This approach suggests that stereotypies can be prevented by either reducing the level of motivation underlying the stereotypy, or by allowing for the expression of more complex behavior and thereby preventing the processes of channeling and sensitization from occurring. PMID- 8226386 TI - Nursing in pigs: communicating needs and distributing resources. AB - When nursing, the sow has a problem to solve different from that of most other mammals because she gives birth to a large number of precocial young. The sow has to incorporate elements into its nursing process that enable it to distribute milk evenly among its young in spite of their being able to fight with and suckle independently of their littermates. This paper described the different features of the nursing process, discussing the functions of a long initial massage before ejection, the grunting of the sow during the first phases of nursing, the postejection massage, and nonnutritional nursings in the context of a process of communication. Through tactile stimulation of the udder by the pigs, the sow releases a number of pituitary, gut, and pancreatic hormones. The physiological response of the sow is suggested to be quantitatively related to the tactile input given by the pigs. Evidence is presented to support the idea that a fault in the communication process disturbs the nursing-suckling interaction and, thus, the transfer of milk from the sow to its pigs. Also, the importance of some environmental factors that affect this process is discussed. PMID- 8226387 TI - Cloning and sequencing of the cDNA encoding goat beta-lactoglobulin. PMID- 8226388 TI - Nucleotide sequence of the goat kappa-casein cDNA. PMID- 8226389 TI - Photolysis of oral bacteria and its potential use in the treatment of caries and periodontal disease. PMID- 8226390 TI - Effect of heat treatment on Listeria monocytogenes and gram-negative bacteria in sheep, cow and goat milks. AB - Sheep milk, compared with cow and goat milk, had a protective effect on Gram negative bacteria and Listeria spp. heated at 65 degrees C in a test-tube method. This effect was not solely due to fat content as cow milk artificially reconstituted to 10% homologous fat was not as protective. Listeria monocytogenes in whole sheep, cow and goat milks at an inoculum level of 1 x 10(6) cfu ml-1 was heated at 68 degrees C for 15 s in the plate pasteurizer and survival was only detected in whole sheep milk after heating. Whole sheep, cow and goat milks containing high levels of L. monocytogenes (1 x 10(6) cfu ml-1) could not survive the current HTST plate pasteurization protocol. PMID- 8226391 TI - Antibacterial activity of Lactobacillus strains isolated from dry fermented sausages. AB - One hundred strains of lactic acid bacteria isolated from dry cured sausages were tested for antagonistic activity against a set of test strains. Nine of 52 strains of Lactobacillus casei and three of 48 strains of Lact. plantarun produced inhibition zones against the indicator species. The substance excreted by Lact. casei CRL 705 was active against Lact. plantarum, Listeria monocytogenes, Staphylococcus aureus and a wide range of Gram-negative bacteria. The activity of the antibacterial compound from Lact. casei CRL 705 was destroyed by papain, trypsin and pepsin, but was resistant to heat (100 degrees C for 20 min), lysozyme and catalase. The agent was produced during the growth cycle and when the concentrated and neutralized supernatant fluid was added to a fresh culture of sensitive cells it produced a rapid inactivation. A decrease in optical density (O.D.) over time, indicative of cell lysis, was also observed. These characteristics allowed us to identify the inhibitory compound as a bacteriocin which we termed Lactocin 705. PMID- 8226392 TI - Construction and detection of bioluminescent strains of Bacillus subtilis. AB - Bioluminescence (lux) genes from Vibrio fischeri and V. harveyi were introduced into Bacillus subtilis on a plasmid vector and by chromosomal integration. The plasmid-bearing strain was highly luminescent and stable under antibiotic selection, but luminescence was lost in the absence of selection and following sporulation and germination. The chromosomally marked strains emitted less light but were found to be stable without the requirement for antibiotic selection and following sporulation and germination. Individual luminescing colonies of both B. subtilis strains could be detected against a high background of non bioluminescent indigenous soil microbial colonies on agar plates using a charge coupled device camera. These bioluminescent Gram-positive strains could be of value in studies concerning the survival and spread of genetically-modified micro organisms in soil environments. PMID- 8226393 TI - Kidney bean lectin-induced Escherichia coli overgrowth in the small intestine is blocked by GNA, a mannose-specific lectin. AB - The reversible and dose-dependent hyperplastic growth of the small intestine and accelerated epithelial cell turnover caused by feeding rats with diets containing kidney bean lectin (PHA) increased the proportion of immature cells on the villi whose membrane and/or cytoplasm contained mainly simple, polymannosylated glycans. These new alpha-linked mannosyl terminals, particularly of the damaged epithelium, facilitated the preferential adherence of opportunistic Escherichia coli with mannose-sensitive Type 1 fimbriae, and other coliforms, to the glycocalyx. Accordingly, the growth of the gut was accompanied by a reversible and PHA dose-dependent overgrowth with E. coli. As expected from their common carbohydrate specificity, the inclusion in the diet of the mannose-specific agglutinin from snowdrop (Galanthus nivalis) bulbs (GNA) significantly reduced the extent of E. coli overgrowth, but abolished neither the growth nor the damage caused by PHA to the small intestine. Thus, GNA and perhaps other mannose specific lectins, especially when used in a preventive mode, can be used to specifically block the proliferation of Type 1 E. coli in the small intestine. PMID- 8226394 TI - Effects of the in vitro fermentation of oligofructose and inulin by bacteria growing in the human large intestine. AB - The in vitro fermentability of oligofructose and inulin was compared with a range of reference carbohydrates by measuring bacterial end-product formation in batch culture. Short chain fatty acid and gas formation indicated that these substrates, which occur naturally in the diet and reach the colon in a largely intact form, were utilized by mixed populations of gut bacteria. Bacterial growth data showed that oligofructose and inulin exerted a preferential stimulatory effect on numbers of the health-promoting genus Bifidobacterium, whilst maintaining populations of potential pathogens (Escherichia coli, Clostridium) at relatively low levels. Pure culture studies confirmed the enhanced ability of bifidobacteria to utilize these substrates in comparison with glucose. Batch culture experiments demonstrated that the growth of Bifidobacterium infantis had an inhibitory effect towards E. coli and Clostridium perfringens. Potentially, an increase in the concentration of these substrates in the diet may therefore improve the composition of the large intestinal microflora and have positive effects on the quality of the Western diet. PMID- 8226395 TI - [Changes and present status of a Japanese National Leprosarium--analysis of smear positive rate and relapse in Hoshizuka-Keiaien between 1972-1991]. AB - Change of clinical features of leprosy in a Japanese National Leprosarium Hoshizuka-Keiaien during 20 years (1972-91) was studied by analyzing clinical records. (1) Skin slit smear positive rate among lepromatous and borderline cases once increased from 16.3% (1972) to 28.8% (1981) and then declined to 3% (1991). (2) Relapse was 4.25 cases per annum among 817 patients (0.52% per annum). Relapse of lepromatous leprosy (0.42% per annum) is decreasing, and borderline or neuritic relapse (0.12% per annum) of previously lepromatous cases is the major feature in recent years. (3) In lepromatous relapse cases, it took 3.5 years in average to become smear negative again, but this duration is shortening in recently relapsed cases. (4) Number of erythema nodosum leprosum is remarkably decreased, but iridocyclitis is still observed. (5) 85% of inpatients in 1991 are classified as clinical cure (Japanese criteria 1989). By the advance of chemotherapy and aging of the inpatients (average age: 68 years), geriatric diseases, instead of leprosy, are becoming major problems in Japanese National Leprosaria. PMID- 8226396 TI - Quantitative determination of PCR products by colorimetric method. PMID- 8226397 TI - Construction of genomic DNA library of Mycobacterium leprae Thai-53 strain, and detection of M. leprae specific genes in the library by polymerase chain reaction. PMID- 8226398 TI - [Reminiscences of my life in the field of leprosy (1)]. PMID- 8226399 TI - Management of community-acquired pneumonia. PMID- 8226400 TI - Effect of morphine or phenobarbital on teicoplanin elimination pharmacokinetics. AB - Elimination kinetics following a single dose of teicoplanin in rats pre-treated with morphine sulphate (MS), phenobarbital sodium (Pb), and normal saline (NS) were determined. A microbioassay was used to measure teicoplanin levels. A significant increase in the total clearance of teicoplanin was found in rats pre treated with MS as compared to controls (P < 0.048). Wide variability was observed in the renal and non-renal clearances of teicoplanin. The mean renal clearance for rats pre-treated with MS, Pb and NS was 0.61 +/- 0.07.mL/min/kg, 0.60 +/- 0.13 mL/min/kg, and 0.46 +/- 0.02 mL/min/kg, respectively; the mean non renal clearance was 0.33 +/- 0.18 mL/min/kg, 0.17 +/- 0.15 mL/min/kg, and 0.08 +/ 0.03 mL/min/kg, respectively. The differences among the groups for renal and non renal clearance were not statistically significant. The mean apparent volume of distribution of teicoplanin at steady state was significantly lower in the Pb-pre treated rats as compared to controls (P < 0.043). The mean half-life for MS-, Pb , and NS pre-treated groups was 8.1 +/- 3.1 h, 5.9 +/- 3.3 h, and 34.6 +/- 20.7 h, respectively. The differences in mean half-life among the groups achieved statistical significance (P < 0.016). The increase in the total clearance of teicoplanin can best be explained by an increase in both renal elimination and hepatic metabolic pathways. PMID- 8226401 TI - Pharmacokinetics and amoebicidal activity of (+-)-(E)-3-(4- methylsulphinylstyryl)-1,2,4-oxadiazole (BTI 2286E) and its sulphone metabolite (BTI 2571E) in the golden hamster, Mesocricetus auratus. AB - BTI 2286E is a 1,2,4-oxadiazole with amoebicidal activity. Three groups of golden hamsters received single doses of BTI 2286E or its sulphone metabolite BTI 2571E as either BTI 2286E 60 mg/kg po or BTI 2571E 60 mg/kg po or BTI 2571E 60 mg/kg ip. Blood samples were collected up to 8 h post-dose and plasma concentrations of BTI 2286E and BTI 2571E were assayed by HPLC. BTI 2286E was rapidly absorbed, extensively metabolized during first pass and rapidly eliminated with a plasma elimination half-life of 1.32 h. Conversion to BTI 2571E was the major pathway of elimination. BTI 2571E had approximately 40% bioavailability after oral administration. After intraperitoneal administration its absorption was slow and prolonged, with an apparent elimination half-life of 2.77 h, considerably longer than the elimination half-life of 0.67 h observed when BTI 2571E was formed as a metabolite, in vivo. The amoebicidal activity of both the compounds was evaluated in the acute hamster hepatic model of amoebiasis. Both BTI 2286E and BTI 2571E were administered as single graded po or ip dose, and their dose-response profiles were characterized. BTI 2571E exhibited poor activity after oral administration (ED50 70 mg/kg) probably due to poor bioavailability, but after intraperitoneal administration its activity (ED50 40 mg/kg) was comparable to that of BTI 2286E after po or ip administration. PMID- 8226402 TI - Absorption of oral ofloxacin after cytotoxic chemotherapy for haematological malignancy. AB - Ten patients with newly diagnosed haematological malignancy were given ofloxacin, 400 mg daily orally, for antibacterial prophylaxis during neutropenia. Before cytotoxic chemotherapy was given, and on three occasions between days 2-3, 5-7 and 8-10 afterwards, blood samples were taken before and 1, 2, 3, 4, 6 and 10 h after administration of ofloxacin. Serum ofloxacin concentrations were measured by HPLC. Mean (range) pharmacokinetic parameters before chemotherapy were: Cmax, 6.1 (2.7-9.1) mg/L; AUC0-2, 7.5 (1.9-11.6) mg.h/L. On days 2-3 post-chemotherapy the values were reduced to: Cmax, 5.0 (3.6-7.3) mg/L; AUC0-2, 5.8 (1.9-7.8) mg.h/L. On days 5-7 the values were: Cmax, 6.1 (4.0-10.4) mg/L; AUC0-2, 8.5 (0.8 16.8) mg.h/L, and by days 8-10 the values were: Cmax, 6.4 (3.3-12.5) mg/L; AUC0 2, 8.6 (0.8-23.1) mg.h/L. There was a statistically significant difference between Cmax pre- and Cmax two to three days post-chemotherapy (paired t-test, P = 0.05). These changes are similar to those previously reported with ciprofloxacin but less marked. PMID- 8226403 TI - In-vitro and in-vivo studies of the decrease of amphotericin B toxicity upon association with a triglyceride-rich emulsion. AB - The effects of amphotericin B associated with a triglyceride-rich emulsion (AB emulsion), shown previously to behave like lymph chylomicrons, were evaluated in vitro and in vivo. Incorporation of amphotericin B to the emulsion was monitored by UV-visible spectrophotometry. Whilst conventional amphotericin B induced a considerable K+ efflux from erythrocytes, AB-emulsion had essentially no effect. In contrast, the K+ efflux from Candida albicans was similar upon incubation either with AB-emulsion or with conventional amphotericin B. Administration to rats showed reduced mortality due to AB-emulsion compared with conventional amphotericin B. Renal toxicity was also decreased upon AB-emulsion treatment, as evaluated by serum urea and creatinine levels. These data suggest that chylomicron-like emulsions could be utilized as vehicles for amphotericin B in antifungal therapy. PMID- 8226404 TI - Pharmacokinetics and tolerance of liposomal amphotericin B in patients. AB - Amphotericin B incorporated into small unilamellar liposomes prepared from soya phosphatidylcholine and cholesterol in a molar ratio of 7:3 was administered to 12 patients with suspected or documented systemic fungal infection. Each patient received 0.1, 0.4 and 1.0 mg/kg dose of liposomal amphotericin B. Liposomal amphotericin B was well tolerated by these patients with no dose-limiting toxicity. Mild to moderate fever with rigors occurred in 3 patients after the higher doses. Peak plasma amphotericin B concentrations, measured by HPLC; were 0.747-1.429 mg/L and 24 h trough concentrations were 0.148-0.363 mg/L. The mean T1/2 beta of amphotericin B was 17.2 h with a mean volume of distribution of 2.285 L/kg. While these pharmacokinetic parameters obtained with this liposomal amphotericin B formulation are comparable to the values obtained after conventional amphotericin B therapy, they are strikingly different from values obtained by other workers using small unilamellar liposomes with either positive or negative charge. Yet in animal experiments the distribution of all formulations are comparable. PMID- 8226405 TI - An open trial of cefoperazone plus sulbactam for the treatment of fever in cancer patients. AB - Cancer patients received cefoperazone plus sulbactam for 673 febrile episodes presumed to be caused by infection. Overall, 415 (76%) of the 545 evaluable episodes responded. There were 213 responses (84%) amongst the 254 fevers of unknown origin and 202 responses (69%) amongst the 291 documented infections. Fifty-one (61%) of the 83 episodes pneumonia and 74 (64%) of the 115 episodes of bacteraemia responded. Only 39 (58%) of the 67 infections caused by Gram-positive bacteria responded compared with 55 (86%) of 64 Gram-negative infections which included seven of eight caused by Pseudomonas aeruginosa. Eighteen (67%) of 27 polymicrobial infections responded to the regimen. Response rates were significantly lower amongst the 125 patients whose neutrophil counts decreased during therapy than amongst the 158 patients whose neutrophil counts increased. Adverse events which were possibly or probably related to antibiotic therapy were observed during 73 of the episodes; the most commonly reported side-effects were diarrhoea and skin rash. Six patients developed a coagulopathy without haemorrhage and two experienced anaphylactic reactions. In this open trial cefoperazone plus sulbactam proved to be an effective regimen for initial therapy of fever in cancer patients. It should be combined with a glycopeptide in those institutions where infections caused by methicillin-resistant staphylococci are frequently encountered. PMID- 8226406 TI - Antibiotic guidelines and antibiotic utilization in Dutch hospitals. AB - In April-May 1991 the availability of antibiotic formularies in Dutch hospitals was analysed as well as data available on antibiotic susceptibility patterns. In addition data on the use of different groups of antimicrobial agents (aminoglycosides, beta-lactam compounds including aztreonam and imipenem, co trimoxazole, vancomycin and fluoroquinolones) were collected by a questionnaire. Thirty-eight formularies were received which were used in 78 hospitals. No formulary was available from 31 hospitals: 15 hospitals did not intend to set up antibiotic guidelines, nine hospitals were using their first formulary and seven used a revised version of the existing one. Fifteen formularies dated from 1987 or earlier. The availability of antibiotic susceptibility patterns ranged from no data at all (n = 14) to extensive data per species, per agent and per ward (n = 5). From 20 hospitals data on antibiotic usage as well as on antibiotic guidelines were obtained. Because several hospitals used the same formulary, 15 different formularies were analysed from 20 hospitals. Fluoroquinolones were used in all hospitals, but were only mentioned in the formularies of six hospitals. The data in the present study underscore the need for a collaborative approach of medical staff, hospital pharmacy and microbiologist in order to maintain a relatively low level of antibiotic resistance in Dutch hospitals. PMID- 8226407 TI - Ciprofloxacin resistance developing during treatment of malignant otitis externa. PMID- 8226408 TI - In-vitro activity of cefepime against bacterial pathogens from hospitalized patients. PMID- 8226409 TI - Susceptibility of Neisseria meningitidis in Israel to penicillin and other drugs of interest. PMID- 8226410 TI - Susceptibility of cefoxitin-resistant isolates of bacteroides to other agents including beta-lactamase inhibitor/beta-lactam combinations. PMID- 8226411 TI - In-vitro selection of fluoroquinolone resistance in Salmonella typhimurium. PMID- 8226412 TI - The effects of arphamenine-A, an inhibitor of aminopeptidases, on in-vitro growth of Trypanosoma brucei brucei. PMID- 8226413 TI - Use of the polymerase chain reaction (PCR) for the detection of aacA genes encoding aminoglycoside-6'-N-acetyltransferases in reference strains and gram negative clinical isolates from two Belgium hospitals. AB - Genes encoding aminoglycoside 6'-N-acetyltransferases, were identified using the polymerase chain reaction (PCR). Four sets of primers delineating DNA fragments of 209 bp, 250 bp, 260 bp and 347 bp, specific for the four known aacA genes, and probes within these fragments, were constructed based on the nucleotide sequences of the aacA genes. The specificity of the primers was evaluated using reference strains encoding various aminoglycoside-modifying enzymes. The primers reacted with their corresponding aacA genes and did not cross-react with genes coding for other aminoglycoside-modifying enzymes. One hundred and sixty-one aminoglycoside resistant clinical isolates showing AAC(6')I activity were tested using the PCR assays. The gene described by Tran Van Nhieu & Collatz (1987) was the most frequently identified aacA gene. One strain of Citrobacter freundii contained two distinct aacA genes. However, in 46% of the strains, the majority being Serratia spp. and Acinetobacter spp. none of the specific amplified DNA fragments for any of the known aacA genes could be detected. PMID- 8226414 TI - Use of the E test for methicillin sensitivity testing. AB - Performance of the E test for methicillin sensitivity testing was investigated using 21 methicillin-resistant and 28 methicillin-sensitive staphylococci. Thirty of the strains had previously given equivocal results with a strip diffusion method. The E test performed well with Mueller-Hinton agar plates incubated at 30 degrees C for 24 h but only after supplementation with 5% NaCl. Testing on agar without salt was necessary for the recognition of methicillin resistance in one strain. We recommend a breakpoint of 4 mg/L and heavy inocula. The E test can be controlled by testing a control strain on the same plate. PMID- 8226415 TI - Comparison of three methods for the determination of the sensitivity of Helicobacter pylori to metronidazole. AB - A comparison of various methods for the determination of the sensitivity of Helicobacter pylori to metronidazole was undertaken. The validity of the agar dilution, the disc diffusion and the Epsilometer (E) test was studied using a total of 86 strains, 16 of which were known to be resistant to metronidazole. All tests were carried out on Mueller-Hinton agar with 5% sheep blood. The results of the disc diffusion and the E-test were highly significantly (P < 0.001) associated with those of the agar dilution test, which was taken as a standard (r = -0.96 and r = 0.96, respectively). Investigation of the accuracy of the repetitive (n = 10) testing ten sensitive and one resistant strains, showed that the disc diffusion test led to a systematic and significant (2p < 0.001) underestimation of the MIC values calculated from the inhibition zone diameters via linear regression. In contrast, the results of the E-test did not differ significantly from those of the agar dilution test. The precision of the agar dilution test was significantly worse (2p < 0.005) than the E-test and the disc diffusion test. Because of its accuracy and significantly better precision, the E test is recommended as the best and simplest method for routine antibiotic sensitivity testing of H. pylori. PMID- 8226416 TI - Chemoprophylaxis for candidosis and aspergillosis in neutropenia and transplantation: a review and recommendations. Working party of the British Society for Antimicrobial Chemotherapy. PMID- 8226417 TI - The activity of piperacillin/tazobactam against clinical isolates collected in 20 UK centres and the design of a disc test for susceptibility testing. AB - The in-vitro activities of piperacillin plus tazobactam at ratios of 4:1, 8:1 and 16:1 were determined against 952 non-copy clinical aerobic bacterial isolates collected from 20 UK centres. Tazobactam enhanced the activity of piperacillin against Enterobacteriaceae, Staphylococcus aureus, Haemophilus influenzae and Moraxella catarrhalis. No enhancement was noted for Pseudomonas aeruginosa, other Pseudomonas spp., streptococci and enterococci. For 95.6% of strains MICs were either the same or only one two-fold dilution different with all the three ratios of tazobactam, and for most of the remaining strains the piperacillin MIC was lowest with the highest proportion of the tazobactam (4:1). The percentage of strains which remained resistant was also lowest with the 4:1 ratio. Regression analysis of MICs versus inhibition zone size with a variety of disc strengths tested on DST agar indicated that for Enterobacteriaceae, and H. influenzae 30 +4 micrograms, and 5 +1 microgram piperacillin/tazobactam discs gave the most reliable results. However, for S. aureus no disc gave good discrimination. Zone sizes obtained on DST and Iso-Sensitest agar were similar, with 96.1% of 1450 paired tests showing agreement to within 3 mm. PMID- 8226418 TI - Susceptibility of alpha-haemolytic streptococci causing endocarditis to benzylpenicillin and ten cephalosporins. AB - A clinical case of streptococcal endocarditis in which the isolate proved susceptible to third- but not first-generation cephalosporins prompted us to examine the susceptibility of 44 alpha-haemolytic streptococci from cases of endocarditis to ten cephalosporins and benzylpenicillin. Twenty per cent of strains were resistant to penicillin, and 20% were tolerant. Cefazolin, cefuroxime and cefpirome were the most active first-, second- and third generation cephalosporins tested. Other first-generation cephalosporins tested compared poorly to cefazolin. Cefotaxime and cefpirome were moderately active against some penicillin-resistant strains. Penicillin tolerance was common in Streptococcus gordonii, but a correlation between tolerance and dextran production could not be confirmed. PMID- 8226419 TI - Antibiotic resistance of faecal Enterobacteriaceae isolated from healthy volunteers, a 15-week follow-up study. AB - From 25 March to 1 July 1991 faecal samples from healthy volunteers of two cities and their rural surroundings, Weert (n = 91) and Roermond (n = 96) were collected weekly and analysed for the presence of Enterobacteriaceae, resistant to ampicillin, ciprofloxacin, nalidixic acid, neomycin, nitrofurantoin, oxytetracycline, sulphamethoxazole or trimethoprim. In total, 682 and 690 samples from Weert and Roermond, respectively, were analysed for the prevalence and the degree of resistance to each antimicrobial agent. The mean prevalence of resistance of the samples from Weert varied from 28% +/- 12 (mean +/- S.D.) for ampicillin to 0.1% +/- 1 (range 0-2%) for ciprofloxacin. For Roermond the prevalence of resistance varied from 41% +/- 7% for sulphamethoxazole to 0% for ciprofloxacin. The high degree of resistance (i.e. > 50% of the faecal flora of one particular individual) varied from 8% +/- 4 for sulphamethoxazole to 2% +/- 2 for trimethoprim for Weert. For Roermond the figures varied from 14% +/- 4 for sulphamethoxazole to 0.3% +/- 1 (range 0-2%) for nalidixic acid. High degrees of resistance were not found for ciprofloxacin and nitrofurantoin in either city. In Weert ampicillin-resistant Enterobacteriaceae were isolated from 69/91 individuals. PMID- 8226420 TI - Comparative study of the bactericidal activity of ampicillin/sulbactam and erythromycin against intracellular Legionella pneumophila. AB - Intracellular bactericidal activity of ampicillin/sulbactam and erythromycin was determined with a human macrophage-like (U-937) cell line infected with Legionella pneumophila. Cell monolayers inoculated with L. pneumophila were treated with erythromycin, ampicillin, sulbactam, or ampicillin/sulbactam during the logarithmic phase of bacterial growth. Intracellular bacterial counts were determined at 2-h intervals for 8 h from the time that antibiotics were added. The number of viable intracellular bacteria increased during this time by 0.9 x log10 cfu/mL (P < 0.05) in the control culture, did not change significantly in the cultures treated with ampicillin or sulbactam, decreased by 0.8 x log10 cfu/mL (P < 0.05) with erythromycin, and decreased by 1.8 x log10 cfu/mL with ampicillin/sulbactam (P < 0.05). The number of cfu/mL was significantly less after incubation with ampicillin/sulbactam than with erythromycin (P < 0.05). Ampicillin/sulbactam appeared to have greater bactericidal activity against intracellular L. pneumophila than erythromycin in this in-vitro model. The bactericidal action of ampicillin/sulbactam was significantly greater than would be expected from the additive effects of ampicillin plus sulbactam, suggesting synergic bactericidal activity. PMID- 8226421 TI - The Garrod Lecture. Mycobacterial diseases in the 1990s. PMID- 8226422 TI - The accumulation of five antibacterial agents in porin-deficient mutants of Escherichia coli. AB - A library of isogenic mutants containing Mud9-induced deletions of the structural and regulatory genes for the porin proteins OmpF and OmpC of Escherichia coli was constructed. The accumulation of norfloxacin, tetracycline, chloramphenicol, cephalothin and cefoxitin was measured with each strain, and shown to be reproducible with low experimental standard deviations, such that the roles of OmpF, OmpC and PhoE in the accumulation of these agents were determined. All data were statistically analysed to determine whether the differences observed between the data for each mutant compared with those for the other mutants and for the wild-type strain were significant. The loss of OmpF reduced accumulation of norfloxacin, tetracycline, cephalothin and cefoxitin by 16-60% compared to the wild-type parent strain, but reduced accumulation of chloramphenicol by < 10%. The loss of OmpC reduced accumulation of cephalothin and cefoxitin by 13 and 34%, respectively, compared to the wild-type parent strain, but had little effect on the accumulation of norfloxacin, chloramphenicol and tetracycline (< 3%). The loss of both OmpF and OmpC (ompR) reduced accumulation of norfloxacin, chloramphenicol, tetracycline, cephalothin and cefoxitin by 36-68%. However, the presence of PhoE in the absence of both OmpF and OmpC, enhanced accumulation to 52-119% of the concentrations of these five agents accumulated by the wild-type strain. These data suggest that OmpF is the preferred route of entry for three of the antibiotics studied, but not for chloramphenicol and tetracycline which utilize both porins equally well. The high levels of accumulation (30-64%) of all five antibiotics in the absence of all major porins suggest that an alternative mechanism(s) of accumulation is available. PMID- 8226423 TI - Postantibiotic effect of aminoglycosides on staphylococci. AB - The postantibiotic effects (PAEs) of amikacin, gentamicin, netilmicin and tobramycin on Staphylococcus aureus and S. epidermidis were determined in vitro by a bioluminescence assay of bacterial ATP. Five strains of S. aureus and two strains of S. epidermidis were exposed for 1 h to varying concentrations of these aminoglycosides. Following removal of the antibiotics by dilution, bacterial regrowth was monitored at hourly intervals. The duration of the PAE increased with increasing aminoglycoside concentration. The mean PAEs for the five S. aureus strains ranged from 5-10 h at clinically achievable aminoglycoside concentrations (16-32 mg/L of amikacin and 4-8 mg/L of gentamicin, netilmicin and tobramycin). The results for one of the strains of S. epidermidis were similar to those observed for the S. aureus strains, while the PAEs on the other less susceptible S. epidermidis strain were shorter (0.5-2.5 h). For comparison, two of the S. aureus strains were exposed for 1 and 2 h to a range of concentrations of dicloxacillin (0.25-32 mg/L); this agent induced a much shorter PAE (0-2.3 h). It may be important to take account of the PAE when designing dosing regimens. PMID- 8226424 TI - Susceptibility of 539 gram-positive and gram-negative anaerobes to new agents, including RP59500, biapenem, trospectomycin and piperacillin/tazobactam. AB - Susceptibilities of 539 Gram-positive and Gram-negative anaerobes were tested by agar dilution against 15 new and existing antimicrobial agents. Organisms included 218 Bacteroides fragilis group strains, 15 non-fragilis group Bacteroides, 130 Porphyromonas/Prevotella, 49 fusobacteria, 50 peptostreptococci, 53 clostridia and 24 Gram-positive non-sporeforming bacilli. Of 412 Gram-negative bacilli, 89% were beta-lactamase-positive, while only two of the Gram-positive strains (both clostridia) produced this enzyme. Using established and preliminary breakpoints, all strains were susceptible to biapenem and imipenem (MIC90s 1 mg/L) and chloramphenicol (MIC90 8 mg/L). Only one of all the strains tested (a Cl. innocuum) was resistant (MIC > 4 mg/L) to RP59500; the latter had MIC90 2 mg/L, while 98% of strains were susceptible to trospectomycin (MIC90 16 mg/L). Ninety-nine per cent of strains were susceptible to piperacillin/tazobactam (MIC90 8 mg/L) compared to 86% to piperacillin (MIC90 > 64 mg/L). Corresponding data for ticarcillin/clavulanate versus ticarcillin were 97% susceptible (MIC90 8 mg/L) compared to 83% (MIC90 > 64 mg/L). Enhancement of the beta-lactam by the inhibitors was only seen in beta-lactamase-producing strains. Amoxycillin and cefoperazone were less often active (36% susceptible, MIC90 > 256 mg/L and 66% susceptible, MIC90 64 mg/L, respectively). Cefoxitin had greater activity than cefotetan (90% susceptible, MIC90 32 mg/L, compared to 72% susceptible, MIC90 > 64 mg/L). Metronidazole was active against 94% of strains (MIC90 4 mg/L). All metronidazole-resistant strains were Gram-positive (75% of non-sporeforming bacilli, 9% of clostridia and 6% of peptostreptococci). Ninety per cent of strains were susceptible to clindamycin (MIC90 4 mg/L). PMID- 8226425 TI - Activity of meropenem and other antimicrobial agents against uncommon gram negative organisms. AB - The in-vitro activity of meropenem and other antimicrobial agents was tested by agar dilution against 113 uncommon Gram-negative pathogens. Both meropenem and ceftriaxone showed the most activity against the test isolates. The potency of meropenem was greater than imipenem against Pasteurella multocida, Eikenella corrodens, Haemophilus parainfluenzae, Moraxella spp., Kingella spp., and Actinobacillus actinomycetemcomitans. Further work is necessary to assess the in vivo activity of meropenem against these Gram-negative isolates. PMID- 8226426 TI - Beta-lactam susceptibility of Haemophilus influenzae strains showing reduced susceptibility to cefuroxime. AB - Minimum inhibitory concentrations of 13 beta-lactam antibiotics (ampicillin, amoxycillin, amoxycillin/clavulanate, imipenem, cefazolin, cefadroxil, cefaclor, cefuroxime, cefotaxime, cefepime, cefpirome, cefpodoxime and cefixime), were determined for 76 strains of beta-lactamase negative Haemophilus influenzae, isolated over a five year period (1985-1990) that gave reduced zones to cefuroxime on disc testing when compared to the control strain H. influenzae NCTC 11931. MIC90 values for all antibiotics (except imipenem) were approximately ten times higher than the MIC90 values for a susceptible control group. Increased resistance was not associated with any particular biotype, although three biotype III strains were highly resistant to imipenem. More than 50% of strains with reduced susceptibility to beta-lactam antibiotics were isolated from patients with chronic respiratory disease. Published data on the sputum concentration of each antibiotic were compared to the MIC90 values obtained for the susceptible and resistant strains. PMID- 8226427 TI - Multicentre survey of the comparative in-vitro activity of piperacillin/tazobactam against bacteria from hospitalized patients in the British Isles. AB - Twenty-nine British and Irish hospitals each collected up to 300 bacterial isolates from in-patients. The organisms were identified by an appropriate API system or, for staphylococci, by their Gram and coagulase reactions. Disc susceptibility tests were performed. Isolates that gave zones < or = 25 mm to piperacillin/tazobactam (75 micrograms + 10 micrograms) discs were sent to a central laboratory for re-examination and determination of MIC, together with a sample of the more susceptible organisms. Results were evaluated for 6724 isolates. Over 95% of the isolates of Escherichia coli, klebsiellae, Proteus mirabilis, Pseudomonas aeruginosa, Haemophilus, Moraxella and Bacteriodes, spp. streptococci, pneumococci and Enterococcus faecalis were susceptible to piperacillin/tazobactam (defined as giving a zone > or = 22 mm to a 75 micrograms + 10 micrograms disc), as were 86% of Acinetobacter spp. and 82% of the Citrobacter, Enterobacter, Morganella and Serratia group. Tazobactam particularly extended the activity of piperacillin against E. coli isolates (96% susceptible cf. 61% to piperacillin alone) klebsiellae (95% cf. 70%), P. mirabilis (99% cf. 86%), and Acinetobacter spp. (86% cf. 53%). Occasional (18%) resistance in Enterobacter, Serratia and Citrobacger spp. was probably caused by stable depression of Class I beta-lactamases, which are inhibited poorly by tazobactam. High resistance frequencies (> 25%) were found for Enterococcus faecium and Xanthomonas maltophilia. Tazobactam potentiated piperacillin against beta lactamase-producing methicillin-susceptible Staphylococcus aureus, but the mode inhibition zone of piperacillin/tazobactam discs was only 26 mm, compared to 38 mm for beta-lactamase-negative isolates. Nevertheless, fewer than 5% of the enzyme producers appeared resistant to 8 + 4 mg/L piperacillin/tazobactam in MIC tests. Similar behaviour was noted for coagulase-negative staphylococci. Amongst the eleven comparator drugs, ceftazidime, gentamicin and ciprofloxacin were as active as piperacillin/tazobactam against most enterobacteria. However, Acinetobacter and Bacteroides spp. and enterococci were resistant to ceftazidime, and Bacteroides spp., enterococci, pneumococci and other streptococci were inherently resistant to ciprofloxacin and gentamicin. Cefuroxime, ampicillin and co-amoxiclav had narrower spectra. Only imipenem showed a consistently wider spectrum and lower frequency of resistance than piperacillin/tazobactam. PMID- 8226428 TI - In-vitro characteristics of glycopeptide resistant strains of Staphylococcus epidermidis isolated from patients on CAPD. AB - The low-level resistance of three clinical isolates of Staphylococcus epidermidis to glycopeptide antibiotics was found to be constitutive, not inducible, and was not increased by passage in the presence of either vancomycin or teicoplanin. There was no loss of resistance on repeated passage in antibiotic-free broth. In contrast, the susceptibility to these antibiotics declined for S. epidermidis NCTC 6513 that been sequentially passaged in either vancomycin or teicoplanin whereas the variants reverted to being susceptible on further passage in antibiotic-free broth. Antibiotic activity was almost completely abolished when cultures of the resistant S. epidermidis strains were exposed overnight to sub MIC concentrations. No evidence of drug-modifying activity was obtained. Experiments of antibiotic-binding activity indicated that the resistant strains exhibited an increased ability to sequester antibiotics which was particularly rapid in stationary phase cultures when most of the antibiotic activity disappeared from the growth medium within 30 min of exposure to the drugs. Teicoplanin was sequestered more efficiently than vancomycin and some loss of activity was also observed when stationary phase cultures of S. epidermidis NCTC 6513 were exposed to glycopeptides. These results suggest that glycopeptide resistant isolates of S. epidermidis are able to bind large amounts of these antibiotics, possibly at sites unassociated with the D-alanyl-D-alanine target, and that teicoplanin is bound more avidly than vancomycin. PMID- 8226429 TI - Intraphagocytic bioactivity of lomefloxacin against Pseudomonas aeruginosa. AB - The kinetics of extracellular bactericidal activity and the intraphagocytic bioactivity of lomefloxacin at concentrations 2 mg/L (1/2 x MIC), 4 mg/L (MIC) and 8 mg/L (2 x MIC) against Pseudomonas aeruginosa were measured, the first in 40% human serum and the second in polymorphonuclear leucocytes (PMNLs), by the colony-counting method. The PMNLs were treated with phenylbutazone to prevent them killing the bacteria during the ingestion period. The kinetics of extracellular killing were biphasic with an initial rapid phase lasting 30 min when lomefloxacin was used at concentrations equal to the MIC and 2 x MIC, during which a decrease of 3 log of cfu/mL occurred. Lomefloxacin showed intraphagocytic activity at the three concentrations assayed. With non-treated PMNLs the percentages of intraphagocytic killing after 30 min of incubation were 93.5, 91.3 and 95.5% with 2, 4 and 8 mg/L, respectively, and 79.8, 65.0 and 93.3% for the same lomefloxacin concentrations with phenylbutazone-treated PMNLs, suggesting a synergic interaction between the antibacterial activity of PMNLs and lomefloxacin. PMID- 8226430 TI - Alterations of host response by a long-term treatment of roxithromycin. AB - Roxithromycin (10 mg/kg) was administered once-daily for seven or 28 days to mice intragastrically and its effects on cytokine synthesis were studied. Administration of roxithromycin for 28 days resulted in increased synthesis of interleukin (IL)-1 and tumour necrosis factor (TNF-alpha) production by macrophages, and the production of IL-2, IL-4 and interferon gamma by splenocytes. In contrast, 7 day administration of roxithromycin did not affect these parameters. Furthermore, clindamycin treatment did not significantly increase the capacity of host cells to produce cytokines, irrespective of the duration of treatment, when compared with the solvent control receiving 0.9% ethanol solution or the untreated controls. In addition, the supernatant of a 7 day culture of splenocytes with roxithromycin showed suppression of the production of TNF-alpha and prostaglandin E2 by mitogen-stimulated macrophages, and this suppressive activity was impaired by a monoclonal antibody to murine IL 4. Thus, this data suggests that roxithromycin may not only enhance the host defence system through increased cytokine synthesis by host cells, but also exhibit anti-inflammatory activity by including an anti-inflammatory cytokine, IL 4. PMID- 8226431 TI - Bactericidal activity of two different dosage regimens of imipenem in an in-vitro dynamic model. AB - The antimicrobial activity of imipenem was tested in an in-vitro model mimicking human serum pharmacokinetics after either 1000 mg im once daily or 500 mg im twice daily. Six recent clinical isolates of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae and Salmonella group B were used as test strains. Our results suggest that a single daily dose of imipenem 1000 mg im exerts an antibacterial action comparable to that obtained with two divided doses of 500 mg. PMID- 8226432 TI - Bactericidal effect of gentamicin peak concentration provides a rationale for administration of bolus doses. AB - A study was made of the influence of peak gentamicin concentration (Cmax) independent of the area under the antibiotic concentration-time curve (AUC) on the bactericidal effect of gentamicin against Escherichia coli NCTC 10418. Bacteria in the lag phase were exposed to in-vitro gentamicin concentration-time profiles which modelled complete profiles determined in vivo in patients after an intravenous bolus (1 min) or infusion (30 min) regimen. An additional in-vitro profile reflected antibiotic removal 30 min after bolus dosing. Bacterial viable cell counts and gentamicin concentrations were measured before and during antibiotic exposure. Both Cmax and AUC were shown to be factors determining antibacterial activity; however Cmax independently represented some 35% of total exposure effect. These findings indicate grounds for preferring bolus intravenous gentamicin dosing with appropriate attention to potential toxicity. PMID- 8226433 TI - Efficacy of ceftriaxone plus tazobactam in a rat model of intraabdominal abscess due to Bacteroides fragilis. AB - Using a rat model of intraperitoneal abscess due to Bacteroides fragilis, we evaluated therapy with the combination of ceftriaxone plus the beta-lactamase inhibitor tazobactam in comparison with ceftriaxone or cefotaxime alone. When treatment was begun five hours after bacterial challenge, final bacterial counts within abscesses at 3.5 days of treatment were as follows (mean +/- S.D., log10 cfu/g): ceftriaxone plus tazobactam, 4.15 +/- 1.25; cefotaxime, 4.77 +/- 1.80; ceftriaxone alone, 5.68 +/- 1.04; untreated controls, 9.14 +/- 1.13. In spite of pharmacokinetic differences between the two drugs, coadministration of tazobactam significantly enhanced activity of ceftriaxone in this model. PMID- 8226434 TI - Chemical disinfection of duck hepatitis B virus: a model for inactivation of infectivity of hepatitis B virus. AB - The susceptibility of duck hepatitis B virus (DHBV) to the virucidal effects of sodium hypochlorite (NaOCl) and sodium dichloroisocyanurate (NaDCC) was compared to hepatitis B virus (HBV) with the aim of using the duck as a model for studying HBV disinfection. Using viral DNA polymerase (DNAP) as a target, inhibition of DNAP activity by chlorine disinfectants was found to be concentration-dependent but independent of contact time. Two minute exposure of minimal effective concentrations of sodium hypochlorite (domestic bleach: 3600 ppm and industrial bleach: 3180 ppm) and sodium dichloroisocyanurate (3000 ppm available chlorine) to DHBV- and HBV-rich plasma totally inhibited DNA polymerase activity. DHBV particles in DHBV-carrier duck plasma (10(4.5) ID50/mL) were treated with these concentrations and inoculated intravenously into 18 one-day old ducklings (six animals/disinfectant). Analysis of plasma (0, 7 and 14 days post-infection) and post-mortem liver (14 days post-infection) by DNA hybridization techniques showed that DHBV DNA was undetectable in samples from all animals inoculated with disinfected virus particles. However, post-inoculation plasma and liver of 18 of 18 control ducklings inoculated with untreated virions were positive for DHBV DNA. These results show for the first time that total inhibition in vitro of hepadnavirus DNA polymerase activity by chemical disinfectants is predictive of inactivation of infectivity in vivo. PMID- 8226435 TI - Nephrotoxicity of vancomycin and aminoglycoside therapy separately and in combination. AB - There is conflict by evidence as to whether therapy with vancomycin plus an aminoglycoside is more nephrotoxic than therapy with either agent alone. Here we report the results of a prospective, non-randomized, open-label study of the incidence of nephrotoxicity in elderly patients who received vancomycin alone (32 patients) or an aminoglycoside alone (67 patients) or in combination (37 patients). The mean (95% confidence limits) incidence of nephrotoxicity, defined as an increase of > or = 44.2 mumol/L in the serum creatinine, was 19% (5-32%) in patients receiving vancomycin alone, 24% (10-38%) in patients receiving vancomycin plus an aminoglycoside, and 12% (4-20%) in patients receiving an aminoglycoside alone (P > 0.05 for all comparisons). The corresponding absolute increases of the serum creatine were 20.3 +/- 23.0 mumol/L (-24.8 to +65.4 mumol/L), 37.1 +/- 53.0 mumol/L (-67.2 to +140.4 mumol/L), and 22.1 +/- 31.8 mumol/L (-40.7 to +94.6 mumol/L). The absolute increase was significantly greater (P < 0.05) in patients receiving vancomycin plus an aminoglycoside than in patients receiving an aminoglycoside alone. A meta-analysis of seven previously published studies combined with our data revealed that the incidence of nephrotoxicity associated with combination therapy is 13.3 +/- 3.1% (7.3-19.4%) greater than therapy with vancomycin alone (P < 0.01) and 4.3 +/- 1.4% (1.6-7.0%) greater than therapy with an aminoglycoside alone (P < 0.05). The clinical relevance of this finding may be limited in that the mean duration of antimicrobial therapy in three of the studies was greater than 21 days. PMID- 8226436 TI - The BSAC: 21 years and beyond. PMID- 8226437 TI - Standardization of Iso-Sensitest agar. PMID- 8226438 TI - A clarithromycin sensitivity survey in the United Kingdom using Stokes' method. PMID- 8226439 TI - Comparison of the synergic activity of aztreonam or tobramycin plus piperacillin or mezlocillin. PMID- 8226440 TI - Antimicrobial resistance amongst enterococci isolated in the United Kingdom: a reference laboratory perspective. PMID- 8226441 TI - Prevalence of ampicillin-resistant Haemophilus influenzae in community-acquired infections in Hong Kong. PMID- 8226442 TI - Equine plasma and blood volumes decrease with dehydration but subsequently increase with exercise. AB - The effects of dehydration and 40 min of exercise at approximately 40% of maximal O2 consumption on plasma volume (PV) and blood volume (BV) were studied in six horses. Horses were exercised while euhydrated (C); 4 h after administration of furosemide (1.0 mg/kg i.v.; FDH), which induced isotonic dehydration; and after 30 h without water (DDH), which caused hypertonic dehydration. Dehydration resulted in decreases of 6.3 and 9.9% for PV and BV, respectively, with FDH and 10.7 and 8.5%, respectively, with DDH. During exercise in C, PV and BV increased by 12.7 and 20.0%, respectively; during exercise with FDH, they increased by 11.7 and 26.1%, respectively; and during exercise with DDH, PV decreased by 1.3% from predehydration values, while BV increased by 18.7%. Hematocrit and total plasma protein concentration rose to higher values throughout exercise in FDH and DDH than in C; plasma [Na+] was higher in DDH than in FDH and C, [Cl-] was higher in DDH and lower in FDH than in C, and [K+] was lower in FDH and DDH than in C through exercise and recovery. From these results, we conclude that increases in PV and BV are normal features of low-intensity exercise in the horse. The increases in BV not only augment O2 carriage but also help maintain circulating volume. These increases can be modified by preexercise dehydration, the nature of which affects the extent of modification. PMID- 8226443 TI - Muscle glycogen storage after prolonged exercise: effect of the glycemic index of carbohydrate feedings. AB - The effect of the glycemic index (GI) of postexercise carbohydrate intake on muscle glycogen storage was investigated. Five well-trained cyclists undertook an exercise trial to deplete muscle glycogen (2 h at 75% of maximal O2 uptake followed by four 30-s sprints) on two occasions, 1 wk apart. For 24 h after each trial, subjects rested and consumed a diet composed exclusively of high carbohydrate foods, with one trial providing foods with a high GI (HI GI) and the other providing foods with a low GI (LO GI). Total carbohydrate intake over the 24 h was 10 g/kg of body mass, evenly distributed between meals eaten 0, 4, 8, and 21 h postexercise. Blood samples were drawn before exercise, immediately after exercise, immediately before each meal, and 30, 60, and 90 min post prandially. Muscle biopsies were taken from the vastus lateralis immediately after exercise and after 24 h. When the effects of the immediate postexercise meal were excluded, the totals of the incremental glucose and insulin areas after each meal were greater (P < or = 0.05) for the HI GI meals than for the LO GI meals. The increase in muscle glycogen content after 24 h of recovery was greater (P = 0.02) with the HI GI diet (106 +/- 11.7 mmol/kg wet wt) than with the LO GI diet (71.5 +/- 6.5 mmol/kg). The results suggest that the most rapid increase in muscle glycogen content during the first 24 h of recovery is achieved by consuming foods with a high GI. PMID- 8226444 TI - Effect of reduced hemoglobin concentration on leg oxygen uptake during maximal exercise in humans. AB - Maximum oxygen uptake (VO2max) is affected by hemoglobin concentration ([Hb]). Whether this is simply due to altered convection of O2 into the muscle microcirculation or also to [Hb]-dependent diffusive transport of O2 out of the muscle capillary is unknown in humans. To examine this, seven healthy volunteers performed four maximal cycle exercise bouts at sea level immediately after 8 wk at altitude (3,801 m, barometric pressure 485 Torr), a sojourn designed to increase [Hb]. The first two bouts were at ambient [Hb] of 15.9 +/- 0.7 g/100 ml breathing 21 or 12% O2 in random order. [Hb] was then decreased to a prealtitude level of 13.8 +/- 0.6 g/100 ml by venesection and isovolemic replacement with 5% albumin in 0.9% saline, and the exercise bouts were repeated. At whole body VO2max, PO2, PCO2, pH, and O2 saturation were measured in radial arterial and femoral venous blood. Femoral venous thermodilution blood flow was determined for calculation of leg VO2. Mean muscle capillary PO2 and muscle diffusing capacity (DO2) were computed by Bohr integration between measured arterial and femoral venous PO2. Averaged over both fractional concentrations of inspired O2, leg VO2 at maximum decreased by 17.7 +/- 4.3% as [Hb] was lowered while leg O2 delivery decreased by 17.5 +/- 2.6% and DO2 decreased by 10.7 +/- 2.7% (all P < 0.05). The relative contributions of decreases in leg O2 delivery and DO2 to the decrease in VO2max were computed to be 64 and 36%, respectively. These findings suggest that [Hb] is an important determinant of O2 diffusion rates into working muscle in humans. Possible mechanisms include 1) dependence of DO2 on intracapillary red blood cell spacing, 2) changes in the total rate of dissociation of O2 from [Hb], and 3) increased red blood cell flow heterogeneity as [Hb] is reduced. PMID- 8226445 TI - Additive effects of age and silent myocardial ischemia on the left ventricular response to upright cycle exercise. AB - To separate the independent effects of age and silent myocardial ischemia on the left ventricular response to aerobic exercise, maximal upright cycle ergometry was performed in three groups: 8 clinically healthy older men [76 +/- 3 (SE) yr] with ischemic electrocardiogram (ECG) and Tl scan responses to prior maximal treadmill exercise (old silent ischemic subjects), 16 age-matched men with normal ECG and Tl scan responses (old controls), and 21 healthy young (33 +/- 1 yr) men (young controls). Although the left ventricular ejection fraction, end-diastolic volume index, and end-systolic volume index were similar in the three groups at rest, with increasing work loads there was a progressive increase in the end diastolic volume index and a blunted decline in end-systolic volume index in the two older groups, which was most apparent in the old silent ischemic subjects. Thus, at peak effort, end-diastolic volume index was largest in old silent ischemic subjects (101 +/- 6 ml/m2), intermediate in old controls (85 +/- 6 ml/m2), and smallest in young controls (67 +/- 3 ml/m2) (P < 0.002); conversely, left ventricular ejection fraction was highest in young controls (85 +/- 2), intermediate in old controls (76 +/- 3), and lowest in the old silent ischemic group (66 +/- 2) (P < 0.001). At exhaustion the peak systolic pressure-end systolic volume index was significantly lower in the silent ischemic group than in young controls (6 +/- 1 vs. 25 +/- 4 mmHg.ml-1 x m-2, respectively; P < 0.001) with the old controls in between (16 +/- 5 mmHg.ml-1 x m-2).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226446 TI - Myocardial substrate uptake in lambs with and without aortopulmonary shunts during strenuous exercise. AB - Increased myocardial fatty acid uptake during acute exercise could adversely affect myocardial O2 consumption in lambs with left-to-right shunts, which would be unfavorable in view of their decreased coronary blood flow reserve. Therefore, we studied myocardial substrate uptake (glucose, lactate, pyruvate, free fatty acids, triglycerides, beta-hydroxybutyrate, and acetoacetate) in 10 7-wk-old lambs with an aortopulmonary left-to-right shunt [61 +/- 3% (SE) of left ventricular output] and 9 control lambs during strenuous treadmill exercise. The hemodynamic reaction to exercise was similar in shunt and control lambs. The peripheral metabolic response to exercise was also similar in the two groups: glucose free fatty acids, and, most prominently, lactate concentrations increased. Myocardial O2 consumption increased but less in shunt than in control lambs because of a smaller increase in heart rate. In both groups myocardial lactate uptake increased substantially at the cost of other substrates, providing the heart with 40% of its oxidative metabolism. Fatty acid uptake was not different between the two groups. In conclusion, our data reveal no essential differences in myocardial substrate uptake between shunt and control lambs during a substantial circulatory load. PMID- 8226447 TI - Partitioning of pulmonary impedance: modeling vs. alveolar capsule approach. AB - Pulmonary input impedance (ZL), transfer tissue impedances (Ztti), and transfer airway impedances (Ztaw) were measured in open-chest dogs and isolated canine lungs by means of small-amplitude pseudorandom oscillations between 0.2 and 21.1 Hz. In the determination of Ztti and Ztaw, local alveolar pressures (PA) sensed in alveolar capsules were used. The global impedances of the airways (Zaw) and tissues (Zti) were estimated by fitting to the ZL data between 0.2 and 4.9 Hz (open-chest dogs) and between 0.2 and 5.9 Hz (isolated lungs) two models based on Hildebrandt's formulations (Bull. Math. Biophys. 31: 651-667, 1969), the parameters of which included airway resistance (Raw) and inertance (Iaw) and tissue damping (GL) and elastance (HL). The tissue parameters of Ztti (Gti and Hti) were also obtained from model fitting, whereas the Ztaw data were evaluated in terms of resistance (Rtaw) and inertance (Itaw). Excellent agreement was found between HL and Hti in both experimental groups and between GL and Gti in the isolated lungs (r > or = 0.999). The damping coefficients were also closely related in the open-chest dogs (r = 0.95), but Gti overestimated GL slightly (by 9%). Raw was underestimated by Rtaw (by 3-33%) and Iaw by Itaw (by 2-16%), depending on the model type and, in the excised lungs, the number of punctures in the capsules. In the case of the airway parameters, the systematic differences were accompanied by lower r values (0.535-0.935), which are explained primarily by the regional variations in PA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226448 TI - Developmental changes in diaphragm contractile properties. AB - The contractile properties of pre- and early postnatal respiratory muscles are incompletely understood. We examined the effects of development on isometric contractile properties, with an emphasis on properties at 37 degrees C. One-day old (n = 10), 3-wk-old (n = 10), and adult (n = 10) rabbits were studied. Isometric contractile properties of costal diaphragm strips were measured in vitro by using direct stimulation. Twitch and maximal, i.e., fused, tetanic force production increased with strip dimension and with age. Maximal tetanic force developed per unit cross-sectional area (stress) was significantly decreased in muscle from 1-day olds, whereas it was greatest in muscle from 3-wk olds. Twitch stress was similar in all three groups. Only when the stimulus duration was prolonged did twitch and fused tetanic force achieve maximal values values for the 1-day-old and 3-wk-old strips, suggesting less effective excitation contraction coupling in those muscles. We conclude that immature rabbit diaphragm has unique isometric contractile properties and stimulus parameter requirements that cannot be deduced from studies using mature diaphragm. PMID- 8226449 TI - Costal diaphragm curvature in the dog. AB - The curvature of the midcostal region of the diaphragm in seven dogs was determined at functional residual capacity (FRC) and end inspiration during spontaneous breathing and mechanical ventilation and at total lung capacity in the prone and supine positions. Metallic markers were attached to muscle fibers on the abdominal surface of the diaphragm, and the dog was allowed to recover from surgery. The three-dimensional positions of the markers were determined by biplane videofluoroscopy. A quadratic surface was fit to the bead positions. The principal axes of the quadratic surface lie nearly along and perpendicular to the muscle fibers. In both the supine and prone positions, the values of the principal curvatures were similar at FRC and end inspiration during spontaneous breathing, when muscle tension and transdiaphragmatic pressure both increase with increasing lung volume, and during mechanical ventilation and passive inflation to total lung capacity, when both decrease relative to their magnitude at FRC. No abrupt change of curvature, which might be expected at the edge of the zone of apposition, was apparent. The curvature along the muscle fiber was 0.35 +/- 0.07 cm-1; the curvature perpendicular to the muscle fiber was much smaller, 0.06 +/- 0.01 cm-1. The costal region of the diaphragm displaces and shortens as lung volume increases, but its shape, as described by its curvatures, does not change substantially. PMID- 8226450 TI - Effects of hypoxia and atrial natriuretic peptide on aldosterone secretion in healthy subjects. AB - To evaluate the inhibitory effect of hypoxia and atrial natriuretic peptide (ANP) on aldosterone secretion, 11 healthy male subjects were infused with 5 ng.kg-1 x min-1 ANP or placebo. The subjects were exposed in a stepwise fashion to incremental hypobaric hypoxia, which decreased arterial oxygen saturation to 79 +/- 2% in the placebo and 84 +/- 2% in the ANP condition (P < 0.05). In the placebo condition, the plasma ANP concentration increased from 13.8 +/- 1.0 to 19.6 +/- 2.3 pmol/l (P < 0.01) at the lowest barometric pressure. Plasma renin activity did not change, whereas the plasma aldosterone levels increased consequent to the increase of plasma adrenocorticotropic hormone (ACTH). Continuous infusion of ANP increased the plasma levels twofold (P < 0.001) and the level of guanosine 3',5'-cyclic monophosphate threefold (P < 0.001). However, the plasma aldosterone concentrations were not different in the two experimental conditions. Administration of supplementary oxygen significantly decreased ACTH to baseline values (P < 0.01) together with a decrease in aldosterone. Free water clearance (P = 0.05) but not sodium excretion (P = NS) increased during continuous ANP infusion. The data indicate that the aldosterone secretion in hypoxia is not inhibited by (patho)physiological plasma ANP levels. The inhibition of aldosterone secretion may well be explained by a direct effect of hypoxia on the adrenal cells. ACTH is a major stimulus of aldosterone secretion in hypoxia, which overrides the natriuretic effect of ANP. PMID- 8226451 TI - Hydroxylation of salicylate by the in vitro diaphragm: evidence for hydroxyl radical production during fatigue. AB - There is increasing evidence that oxygen-derived free radicals produced during strenuous work by the diaphragm may contribute to diaphragm fatigue and/or injury. However, the precise identity of these oxygen radicals remains unknown, inasmuch as oxygen free radicals are extremely short lived and their detection in biologic systems is quite difficult. There is recent evidence that the salicylate trapping method may be a useful means of monitoring tissue production of hydroxyl radical (.OH). This method is predicated on the fact that salicylate's phenolic ring can be attacked by .OH at the 3 or 5 position to yield 2,3- or 2,5 dihydroxybenzoic acid (DHB). These metabolites are stable and can be identified by high-performance liquid chromatography (HPLC) coupled with electrochemical or ultraviolet detection. To test the hypothesis that hydroxylated salicylates are produced during diaphragm fatigue, we exposed in vitro rat diaphragm strips to a physiological saline solution containing 2.0 mM sodium salicylate for approximately 15 min. The solution was then removed, and the strips were fatigued (20 Hz, 200-ms train duration, 1 train/s) via phrenic nerve stimulation for 30 s 10 min. The diaphragm strips were subsequently homogenized, and the homogenate was analyzed by HPLC coupled with ultraviolet detection. Levels of 2,3-DHB were significantly higher in fatigued than in control nonfatigued strips. There was also a significant correlation between the amount of 2,3-DHB in the fatigued muscle and the accumulated tension-time product developed during fatigue. 2,5-DHB was not consistently identified in control or experimental strips.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226452 TI - Upper airway obstruction induced by negative-pressure ventilation in awake healthy subjects. AB - Negative-pressure ventilation (NPV) induces sleep-related upper airway obstruction. However, the precise mechanism and site of upper airway obstruction during NPV have not been worked out. We studied seven awake healthy volunteers (23-30 yr old) in an Emerson tank respirator. Subjects had the head outside the iron lung and breathed through a pneumotachograph, which yielded the airflow (V) signal. Supraglottic pressure (Psg) was measured with a catheter with the tip at the retroepiglottic level. Diaphragmatic electromyograms (EMGdi) were obtained from an esophageal bipolar electrode. Tidal volume was measured with an inductance plethysmograph. Measurements were done at -10, -20, and -30 cmH2O. At each pressure run subjects were asked to repeatedly relax or to actively breathe in phase with the respirator. Subjects had been previously trained to relax during NPV. During the relax runs there was no EMGdi activity. Stridor or wheezing occurred in all seven subjects during the relax runs but not during the active runs. Two patterns were associated with NPV during relax runs. One pattern was decreases in both V and Psg followed by zero values of these indexes, which corresponded to an inspiratory narrowing and closure of the glottis. These changes were visualized by fiber-optic bronchoscopy in one subject. The second pattern was a decrease in V and increase in Psg, which corresponded to an inspiratory supraglottic obstruction. In five subjects a supraglottic pattern was observed, whereas in two subjects glottic closure was seen. We conclude that muscular relaxation during NPV produces a decrease in the caliber of the upper airways at the glottic or supraglottic level. An uncoupling of upper airway muscle activity and the diaphragm might be the mechanism responsible for these changes. PMID- 8226453 TI - Effects of acute passive smoking on exercise-induced bronchoconstriction in asthmatic children. AB - This study was designed to investigate the acute effects of environmental tobacco smoke (ETS) in children with mild asthma during rest and exercise. We studied 13 children [8 males, 5 females; mean age 10 (range 8-13) yr; mean forced expired volume in 1 s (FEV1) 93% (range 82-108%) of predicted] with exercise-induced bronchoconstriction [46 +/- 4% (SE) fall in FEV1 after exercise during cold air breathing]. Children were exposed to ETS (20 ppm carbon monoxide) or ambient air (AA) for 1 h. During the first 54 min of exposure, children were at rest, and during the last 6 min they exercised on a bicycle ergometer (2 W/kg body wt). Spirometry was performed before and during exposure and after exercise. Respiratory symptoms were recorded before and after exposures. In seven children the experiments with AA and ETS were done in duplicate. FEV1 between 5 and 54 min of exposure at rest decreased by 3.2 +/- 0.8% (SE) during AA and by 7.2 +/- 2.3% during ETS exposure compared with preexposure values; the difference between AA and ETS was statistically significant (P = 0.04). The drop in FEV1 was achieved within 5 min and did not change with ongoing exposure. Analysis of individual data revealed that the mean changes during ETS were mainly effected by three children with a significant fall and one child with a significant improvement in FEV1 (P < 0.05). Maximum postexercise fall of FEV1 was 25 +/- 4% after AA and 24 +/- 3% after ETS, which did not differ significantly. Upper and lower respiratory tract symptoms were not significantly different between exposures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226454 TI - Phases of metabolism during progressive exercise to fatigue in human skeletal muscle. AB - The purpose of this study was to assess noninvasively both the "oxidative potential" and the sequence of metabolic events that occur during the transition from rest to fatigue in an isometrically exercising muscle. 31P-magnetic resonance spectroscopy was used to obtain continuous measures of intracellular phosphocreatine (PCr), inorganic phosphate (Pi), and proton concentration ([H+]) in the tibialis anterior muscle of eight healthy human volunteers during a progressive isometric exercise protocol. The exercise protocol consisted of 2-min stages, with a duty cycle consisting of 4 s of contraction and 6 s of relaxation, beginning at 10% of the force from an initial maximal voluntary contraction (MVC) and increasing by 10% to a final load of 80% MVC. An MVC was performed at the beginning of each stage; a decrease in MVC indicated fatigue. The initial linear slope of the relationship between force and Pi/PCr was used as an index of oxidative potential for the muscle. This initial slope ranged from 85 to 167% MVC/(Pi/PCr), indicating substantial variability of oxidative potential in these subjects. The changes in Pi/PCr and [H+] over time were best described with a bilinear fit of the individual data. The inflection point for each fit was defined as the point at which the slopes intersected. The Pi/PCr inflection point occurred at a similar value of Pi/PCr in all subjects [0.47 +/- 0.04 (SE)]. The Pi/PCr inflection point occurred significantly earlier (7.8 +/- 0.7 min) than the [H+] inflection point (9.3 +/- 0.4 min, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226455 TI - Central cardiovascular pressures during graded water immersion in humans. AB - Thermoneutral (34.9 degrees C) water immersion (WI) was conducted with 12 upright seated normal males at four consecutive water levels (5-10 min each): knee (reference), xiphoid process, fourth intercostal space, and sternoclavicular notch. Thereafter, water was let out of the tank and the experiment was repeated from the neck to the knees at the same levels. Arterial pulse pressure (PP), central venous pressure (CVP), and transmural CVP (TCVP = CVP - esophageal pressure; n = 4) gradually increased with increasing water levels (P < 0.05). Heart rate (HR) decreased at WI to the xiphoid process (P < 0.05) and thereafter remained at this level, whereas mean arterial pressure remained unchanged. There was a closer linear correlation between HR and PP (r = -0.35, P < 0.01) than between HR and CVP (r = -0.13, P > 0.05). Furthermore, there was a significant positive linear correlation between CVP and TCVP (r = 0.83, P < 0.01). We conclude that WI in humans induces an increase in cardiac filling pressures with an increase in PP and a consequent decrease in HR. Furthermore, changes in CVP accurately reflect changes in cardiac distension (TCVP) during WI. PMID- 8226456 TI - Fatigue-induced alterations in Ca2+ and caffeine sensitivities of skinned muscle fibers. AB - The purpose of this investigation was to examine the Ca2+ and caffeine sensitivities of skinned skeletal muscle fibers after fatigue. Single frog semitendinosus fibers were chemically skinned in either a rested state or after tetanic contractions (80 Hz, 100 ms) evoked at 2 s-1 for 5 min. This protocol reduced tetanic force to 1.8 +/- 0.2% of control. Maximal Ca(2+)-activated force (F0, 20 degrees C) was not significantly different between rested and fatigued fibers. However, the concentration of Ca2+ required to evoke 50% of F0 was significantly lower in the fatigued fibers (1.80 +/- 0.18 vs. 1.33 +/- 0.16 microM; P < 0.05), an effect that persisted as the skinned fiber was allowed to incubate in the relaxing solution for > 90 min. The addition of caffeine (25 mM) after Ca2+ loading of the sarcoplasmic reticulum (SR) for periods of 5-30 s (0.25 microM free [Ca2+]) evoked smaller contractures in fatigued fibers than in rested fibers. However, when the loading period was prolonged (60-240 s), force developed after caffeine application was not significantly different between conditions. This suggests that the rate, but not the maximal capacity of Ca2+ loading by the SR, is reduced by fatigue. After Ca2+ loading (120 s), the minimal caffeine concentration required to evoke a contracture in fatigued fibers (5.7 +/ 0.3 mM) was significantly greater than that of control fibers (3.1 +/- 0.4 mM), an effect that persisted with prolonged incubation of the skinned fibers. In addition, the rate of force increase in response to 8 mM caffeine was reduced in fatigued fibers by 41%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226457 TI - Changes in hormonal concentrations after different heavy-resistance exercise protocols in women. AB - Nine eumenorrheic women (age 24.11 +/- 4.28 yr) performed each of six randomly assigned heavy-resistance protocols (HREPs) on separate days during the early follicular phase of the menstrual cycle. The HREPs consisted of two series [series 1 (strength, S) and series 2 (hypertrophy, H)] of three protocols, each using identically ordered exercises controlled for load [5 vs. 10 repetitions maximum (RM)], rest period length (1 vs. 3 min), and total work (J) within each three-protocol series. Blood measures were determined pre-, mid- (after 4 of 8 exercises), and postexercise (0, 5, 15, 30, 60, 90, 120 min and 24 and 48 h). In series 1, a significant (P < 0.05) reduction in growth hormone (GH) was observed at 90 min postexercise for all three protocols. In series 2, the 10-RM protocol with 1-min rest periods (H10/1) produced significant increases above rest in GH concentrations at 0, 5, and 15 min postexercise, and the H10/1 and H5/1 protocols demonstrated significant reductions at 90 and 120 min postexercise. Cortisol demonstrated significant increases in response to the S10/3 protocol at 0 min, to the H10/1 protocol at midexercise and at 0 and 5 min postexercise, and to the H5/1 protocol at 5 and 15 min postexercise. No significant changes were observed in total insulin-like growth factor I, total testosterone, urea, or creatinine for any of the HREPs. Significant elevations in whole blood lactate and ammonia along with significant reductions in blood glucose were observed. Hormonal and metabolic blood variables measured in the early follicular phase of the menstrual cycle varied in response to different HREPs. The most dramatic increases above resting concentrations were observed with the H10/1 protocol, indicating that the more glycolytic HREPs may stimulate greater GH and cortisol increases. PMID- 8226458 TI - Detection of tracheal stenosis by frequency analysis of tracheal sounds. AB - To develop a simple noninvasive method for detecting tracheal stenosis, tracheal sounds were analyzed using fast-Fourier transform. The subjects were all female and included 5 normal volunteers and 13 patients with tracheal stenosis mostly secondary to thyroid cancer (11 extrathoracic and 2 intrathoracic lesions). Tracheal sounds were recorded during spontaneous breathing and were digitized with an analog-to-digital converter. Pulmonary functions, including forced expiratory volume in 1 s (FEV1) expressed as percentage of vital capacity, peak expiratory flow rate (PEFR), the ratio of FEV1 to PEFR (Empey's index), and the ratio of expiratory to inspiratory flow rates at 50% vital capacity, were measured. A computed tomography scan was used to obtain the tracheal minimum cross-sectional area. Whereas PEFR demonstrated a weak correlation with the stenotic area, FEV1%, Empey's index, and the ratio of expiratory to inspiratory flow rates at 50% vital capacity did not. The power of the fast-Fourier transform spectrum of normal tracheal sounds decreased as the frequency increased up to 500 Hz. A small spectral peak was observed at approximately 1 kHz. Patients with significant tracheal stenosis demonstrated an increase in the peak spectral power at approximately 1 kHz and in the mean spectral power from 600 to 1,300 Hz in their tracheal sounds. In patients with extrathoracic lesions, the peak and mean spectral powers correlated well with the area of the stenosis as defined by computed tomography scan. In patients with intrathoracic lesions, abnormalities in the pulmonary functions as well as tracheal sound spectra appeared more evident despite milder stenoses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226459 TI - Lack of importance of the superior laryngeal nerves in citric acid cough in humans. AB - The relative importance of laryngeal afferents in the cough reflex in humans is unknown. This study was designed to investigate the importance of superior laryngeal nerve afferents in the cough reflex induced by inhaled nebulized citric acid in awake humans. Nine healthy volunteers had their cough thresholds to inhaled nebulized citric acid measured after superior laryngeal nerve conduction blockade and after a sham nerve block. Of the nine subjects, four showed no change in cough threshold after superior laryngeal nerve anesthesia, three showed increased cough threshold after nerve block compared with no block, and two showed decreased cough threshold after nerve block. The geometric means of the cough thresholds for the nerve block vs. sham block tests were 16 +/- 13 (SD) and 15 +/- 8% citric acid, respectively. There was no statistically significant difference (Wilcoxon signed-rank test) between the cough thresholds with and without superior laryngeal nerve block (P > 0.05). We conclude that, in the awake human, superior laryngeal nerve afferents do not play a necessary role in initiation of citric acid-induced cough. PMID- 8226460 TI - NG-monomethyl-L-arginine does not restore loss of hypoxic pulmonary vasoconstriction induced by TNF-alpha. AB - Tumor necrosis factor-alpha (TNF-alpha) causes systemic hypotension, pulmonary vasodilation, and loss of hypoxic pulmonary vasoconstriction. NG-monomethyl-L arginine (L-NMMA) inhibits nitric oxide (NO) production and prevents some systemic manifestations of TNF-alpha. We tested using an isolated perfused canine lobe whether NO also mediates the pulmonary vascular effects of TNF-alpha. Total resistance (RT) was measured during control and hypoxic ventilation over a 90-min period in six control lobes, five lobes treated with TNF-alpha (250 micrograms), six lobes treated with L-NMMA (200 mg), and five lobes treated with L-NMMA (200 mg) + TNF-alpha (250 micrograms). In the control lobes RT increased (P < 0.02) from 0.0474 +/- 0.0105 to 0.0677 +/- 0.0133 cmH2O.ml-1 x min during normoxic and hypoxic ventilation, respectively. RT decreased (P < 0.05) from a baseline of 0.0593 +/- 0.0133 to 0.0449 +/- 0.0176 cmH2O.ml-1 x min 30 min after TNF-alpha administration and did not further change during hypoxic ventilation (0.0475 +/- 0.0107 cmH2O.ml-1 x min). L-NMMA pretreatment did not prevent the TNF-alpha induced loss of hypoxic pulmonary vasoconstriction, with values of RT unchanged from normoxic (0.0541 +/- 0.0067 cmH2O.ml-1 x min) to hypoxic (0.0545 +/- 0.0078 cmH2O.ml-1.min) ventilation (P > 0.10) in the L-NMMA + TNF-alpha group after TNF alpha administration. We conclude that NO is not the mediator responsible for the acute pulmonary vascular effects of TNF-alpha. PMID- 8226461 TI - Influence of sleep on alae nasi EMG and nasal resistance in normal men. AB - The influence of sleep on the upper airway musculature varies considerably, with some muscles maintaining their activity at waking levels and others falling substantially. The influence of sleep on the alae nasi (AN), a dilator muscle of the nasal airway, has been minimally studied to date. Thus we determined the effect of non-rapid-eye-movement (NREM) sleep on the AN electromyogram and its relationship to nasal resistance (Rn) in nine normal supine males. Phasic inspiratory AN activity decreased from 20 +/- 6 arbitrary units during wakefulness to 5 +/- 1 arbitrary units (P < 0.001) at the onset of stage 2 NREM sleep and remained unchanged for two subsequent hours of NREM sleep. However, the Rn at the onset of NREM sleep remained similar to awake values (5.7 +/- 0.9 cmH2O.l-1 x s) and increased only after 1 h of NREM sleep (8.6 +/- 1.7 cmH2O.l-1 x s, P < 0.05), thus demonstrating little relationship to AN activity. We conclude that Rn increases slightly after 1 h of sleep, whereas AN activity decreases at stage 2 sleep onset. Thus AN activity has little influence on Rn during sleep. PMID- 8226462 TI - Hindlimb and lung lymph flows during prolonged exercise. AB - We performed experiments to determine the effect of 2h of exercise on hindlimb lymph flow (QL) and protein concentration in sheep. We compared these results with the lung QL response to long-term exercise. Eleven sheep with catheters in an efferent duct of a prefemoral lymph node and 12 sheep with chronic lung lymph catheters exercised at 2.5-3.0 km/h for up to 2h (lung lymph: range 45-120 min, mean 80 min;hindlimb lymph: range 75-120 min, mean 110.5 min). Cardiac output approximately doubled. Pulmonary vascular resistance decreased by 42%, and systemic vascular resistance decreased by 35%. There were small increases in calculated pulmonary microvascular and arterial pressures. During steady-state exercise, lung QL doubled and the lung lymph-to-plasma protein concentration ratio decreased by 16%. There was an immediate fivefold increase in hindlimb QL, and the hindlimb lymph-to-plasma protein concentration ratio decreased by 26%. Hindlimb QL decreased to a constant 130% above baseline during the last 30 min of exercise. We conclude that the marked increase in hindlimb QL early in exercise is secondary to a massaging effect in working muscles. The steady-state increases in QL toward the end of the exercise period in both lung and hindlimb are secondary to both increased surface area and pressure in the pulmonary and systemic microvascular circulations. Our data suggest that in the lung the major factor determining QL is increased vascular surface area. PMID- 8226463 TI - Diaphragm reinnervation by laryngeal motoneurons. AB - Inspiratory activity of the paralyzed diaphragm was restored by reinnervation with brain stem laryngeal motoneurons. In 10 anesthetized cats, the right recurrent laryngeal nerve (RLN) was cut and anastomosed to the distal stump of either one or both roots (C5-C6) of the ipsilateral phrenic nerve. Three to four months later, reinnervation was assessed under deep anesthesia by the reappearance in the paralyzed diaphragm of 1) direct electromyographic (EMG) responses after electrical stimulation of the RLN and 2) spontaneous inspiratory bursts. Serial radiography, performed on five animals, revealed diaphragmatic excursions of comparable amplitude on the normal and reinnervated sides. Six to twelve months after anastomosis, laparotomy (performed under Nembutal anesthesia) allowed inspection and EMG recording of the spontaneous inspiratory contractions of the reinnervated areas and their sustained responses to tetanic RLN stimulation. Inspiratory discharges showed a ramplike recruitment similar to that of the normal diaphragm. Although the RLN contains a number of expiratory axons, multiple-site recordings disclosed expiratory EMG discharges only once. Histological analysis confirmed the substitution of phrenic axons by regenerating RLN fibers. PMID- 8226464 TI - Transient changes in muscle high-energy phosphates during moderate exercise. AB - The purpose of this study was to use 31P-nuclear magnetic resonance spectroscopy to examine changes in wrist flexor muscle metabolism during the transitions from rest to steady-state exercise (on-transient) and back to rest (off-transient). Five healthy young males (mean age 25 +/- 2 yr) performed a series of square-wave exercise tests, each consisting of 5 min of moderate-intensity work followed by a 5-min recovery period. The subjects repeated this protocol six times, and each individual's results were pooled before analysis. ATP and intracellular pH did not change significantly during exercise or recovery. Phosphocreatine (PCr) declined progressively at the onset of exercise, reaching a plateau after approximately 2 min. A reciprocal increase in Pi occurred during the onset of exercise. During the recovery period PCr was resynthesized, whereas Pi returned to resting levels. The data were plotted as a function of time and fit with both first- and second-order exponential growth or decay models; however, the second order model did not significantly improve the fit of the data. Time constants for the first-order model of the on- and off-transient responses for both PCr and Pi were approximately 30 s. These values are nearly identical to the time constants for oxygen consumption during submaximal exercise that have been reported previously by several authors. The results of this study show that the metabolism of muscle PCr during steady-state exercise and recovery can be accurately described by a monoexponential model and, further, suggest that a first-order proportionality exists between metabolic substrate utilization and oxygen consumption. PMID- 8226465 TI - Contribution of hyperoxia to reduced pulmonary function after deep saturation dives. AB - Pulmonary function was measured before and after a 28-day saturation dive to a pressure of 0.25 MPa in eight subjects. PO2 was 40 kPa, with periods of 75 kPa for 2 h every 2nd day during the first 14 days, 50 kPa for the next 12 days, and a gradual fall to 21 kPa over the last 2 days in decompression. A 28-day saturation dive with six subjects to a pressure of 0.15 MPa and a PO2 of 21 kPa was used as control. The measurements included static and dynamic lung volumes and flows, transfer factor for carbon monoxide (TLCO), and a cycle ergometer exercise test. There was a significant reduction in TLCO of 9.8 +/- 6.0% (P < 0.001) after the dive when values were corrected for hemoglobin concentration changes. Effective alveolar volume was unchanged. There was a reduction in forced midexpiratory flow rate of 9.8 +/- 7.0% (P < 0.01), but forced vital capacity and forced expired volume in 1 s were unchanged. Peak oxygen uptake was reduced by 10.1 +/- 5.3% (P < 0.001). There were no significant changes in any of the lung function variables after the control dive. Exposure to raised PO2 contributes significantly to the changes in pulmonary function that have been reported after deep saturation dives to pressures of 3.1-4.6 MPa with a similar profile of oxygen exposure. TLCO is apparently a more sensitive index than vital capacity for oxygen toxicity. PMID- 8226466 TI - Muscle sympathetic nerve responses to graded leg cycling. AB - The aim of this study was to clarify the relationship between sympathetic outflow to skeletal muscle and oxygen uptake during dynamic exercise. Muscle sympathetic nerve activity (MSNA) was recorded from the right median nerve microneurographically in eight healthy volunteers during leg cycling at four different intensities in a seated position for a 16-min bout. Work loads selected were 20, 40, 60, and 75% of maximal oxygen uptake (VO2max). Heart rate and blood pressure were measured during each exercise test. MSNA burst frequency was suppressed by 28% during cycling at 20% VO2max (23 vs. 33 bursts/min for control). Thereafter, it increased in a linear fashion with increasing work rate, with a significantly higher burst frequency during 60% VO2max than the control value. Both heart rate and mean blood pressure rose significantly during 20% VO2max from the control value and increased linearly with increased exercise intensity. During light exercise, MSNA was suppressed by arterial and cardiopulmonary baroreceptors as a result of the hemodynamic changes associated with leg muscle pumping. The baroreflex inhibition may overcome the muscle metaboreflex excitation to induce MSNA suppression during light exercise. These results suggest that during light exercise MSNA is inhibited, perhaps due to loading of the cardiopulmonary and arterial baroreflexes, and that during heavier exercise the increase in MSNA occurs as muscle metaboreflexes are activated. PMID- 8226467 TI - Effects of exercise intensity and duration on norepinephrine spillover and clearance in humans. AB - During dynamic exercise, blood flow to exercising muscle is closely matched to metabolic demands. This is made possible by metabolic vasodilation, vasoconstriction in inactive vascular beds, and a rise in cardiac output. The sympathetic nervous system plays an important role in regulating this exercise response. In this study, we used steady-state infusions of tritiated norepinephrine ([3H]NE) to determine the magnitude and time course of the arterial NE spillover response to sustained upright bicycle exercise at low (n = 11) and moderate-to-high (n = 14) exercise intensity (25 and 65% of maximum work load, respectively) in normal young subjects. In addition, we sought to examine whether exercise was associated with a change in NE clearance. During 30 min of low-level exercise, arterial NE spillover increased from 1.45 +/- 0.13 to 3.14 +/ 0.30 nmol.min-1 x m-2 (P < 0.01) and appeared to plateau at 20-30 min of exercise; NE clearance remained unchanged. During 20 min of moderate-to-high intensity exercise, we found a substantial and progressive rise of arterial NE spillover from 2.15 +/- 0.27 to 13.52 +/- 1.62 nmol.min-1 x m-2 (P < 0.01). NE clearance decreased from 0.91 +/- 0.05 to 0.80 +/- 0.05 l.min-1 x m-2 (P < 0.05). These data suggest that, during dynamic exercise, sympathetic nervous system activity is related to exercise intensity, and there appears to be an interaction between the effects of exercise intensity and duration on NE spillover. In addition, at moderate-to-high exercise intensity, a small decrease of NE clearance contributes to the rise in plasma NE. PMID- 8226468 TI - Respiratory load compensation. II. Role of the cerebellum. AB - Effects of inspiratory tracheal occlusion (TO) on respiratory duration (inspiratory and expiratory duration), ventilation, and the peak integrated diaphragm electromyographic (integral of EMGdi) response were tested in 16 anesthetized cats before and after decerebellation with and without vagal input. The same protocols were repeated in the decerebrate preparation. Decerebellation did not significantly affect the baseline or the loaded values [tracheal occlusion (TO)] for respiratory duration, tidal volume, or magnitude of the integral of EMGdi response. Vagal blockade eliminated the load-compensating responses in the intact and the decerebrate preparation. However, vagal blockade in concert with decerebellation resulted in a significant (P < 0.05) reversible inhibition of the peak integral of EMGdi response during inspiratory TO. This suggests that removal of vagal and cerebellar influences during loaded breathing unmasked inhibitory inputs to the respiratory pattern generator. With vagus intact, decerebellation before or after decerebration abolished the attenuation of the peak integral of EMGdi response to TO observed with decerebration alone. We conclude that the cerebellum does play a role in determining the pattern of the respiratory response to TO. This influence may be direct and/or indirect via interaction with information emanating from suprapontine, vagal, and nonvagal sources. PMID- 8226469 TI - Respiratory load compensation. III. Role of spinal cord afferents. AB - In a previous study, we reported that inspiratory tracheal occlusion (TO) significantly inhibited the motor drive to the diaphragm in a decerebellated bilaterally vagotomized preparation (J. Appl. Physiol. 75:675-681, 1993). The hypothesis to be tested in the present study was that respiratory muscle afferents activated by inspiratory TO provided the inputs responsible for the observed inhibition. Adult cats were anesthetized, tracheotomized, and instrumented with diaphragm electromyographic (EMGdi) recording electrodes. The cerebellum, vagi, and dorsal spinal cord (C2-T2) were surgically exposed. Inspiratory TO was applied before and after cold blockade of the dorsal cord (C6) or dorsal root (C3-6) transection in the intact and decerebellated vagotomized cat. Respiratory timing (inspiratory and expiratory duration) was determined from the EMGdi record, and the peak integrated EMGdi (integral of EMGdi) response was used as an index of respiratory motor drive. Our results showed that 1) cold blockade at the dorsal C6 level in an intact preparation significantly increased the peak of the integral of EMGdi response to TO and was reversible upon rewarming; 2) as previously reported, decerebellation coupled with bilateral vagotomy significantly decreased the peak integral of EMGdi response to TO with no effect on timing; 3) cold blockade (-1 degree C) of the dorsal cord at C6 significantly attenuated this inhibition, and subsequent dorsal rhizotomy at C3-6 completely abolished this inhibition; and 4) decerebellation, cold blockade of the dorsal cord (C6), and dorsal rhizotomy (C3-6) did not significantly affect baseline values in bilaterally vagotomized cats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226470 TI - Influence of the timing of fluid ingestion on temperature regulation during exercise. AB - The purpose of this investigation was to determine whether the timing of fluid ingestion affects thermoregulation during exercise-heat stress. On four occasions, seven endurance-trained cyclists [age 25 +/- 2 (SE) yr, body weight 70.5 +/- 3.3 kg, maximal O2 uptake (VO2max) 4.69 +/- 0.11 l/min] performed 140 min of cycle ergometer exercise at 62-66% of VO2max in a hot environment (33 degrees C dry bulb, 51% relative humidity, wind speed 2.5 m/s). The subjects drank 1,173 +/- 44 ml of a carbohydrate-electrolyte beverage after 0 min (D0), 40 min (D40), or 80 min (D80) of exercise or consumed the same total volume in small aliquots throughout exercise (DT). The exercise-heat stress resulted in calculated sweating rates of approximately 1,200 ml/h and a body weight loss of 2.9 +/- 0.1% after 140 min of exercise. After fluid intake in the D0, D40, and D80 trials, there was a time period (approximately 40 min) in which the increases in serum osmolality and sodium concentration and the reduction in blood volume were attenuated. During that same time period, there was an attenuated rise in esophageal temperature (Tes; P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226471 TI - Lung volume changes during relatively fluent speech in stutterers. AB - We investigated breathing patterns in stutterers during relatively fluent speech and compared these with normal subjects for similar speech tasks. Rib cage and abdominal displacements and esophageal, gastric, and transdiaphragmatic pressures provided indexes of diaphragmatic, rib cage, and abdominal muscle contraction. We found that stutterers spoke either at substantially higher or lower lung volumes than normal subjects, confining their speech to the inspiratory capacity or expiratory reserve volume. During spontaneous speech, stutterers did not cross functional residual capacity (FRC) for most breaths. In addition, stutterers used several different motion pathways from breath to breath. At high lung volumes stutterers used the diaphragm to provide inspiratory braking. At lung volumes below FRC stutterers recruited their abdominals. This contrasted with normal subjects who spoke in the middle part of the vital capacity and who recruited inspiratory and expiratory rib cage muscles above and below FRC, respectively. Breath sizes were log-normally distributed in stutterers compared with a gaussian distribution in normal subjects (P < 0.001). During reading, stutterers tended to cross FRC (P < 0.01), used very similar initiation lung volumes from breath to breath (P < 0.001), and used similar motion pathways to achieve deflation. We conclude that stutterers sustain fluency by speaking at abnormally high or low lung volumes and that this may account for the different muscle patterns observed in stutterers compared with normal subjects. PMID- 8226472 TI - Perfusion of alveolar septa in isolated rat lungs in zone 1. AB - The combination of high inflation and low vascular pressures in zone 1 lungs is assumed to collapse alveolar vessels, making them inaccessible to vascular liquid. To test this assumption, we perfused isolated rat lungs in zone 1 (n = 5) with fluorescent albumin solution (inflation pressure = 25 cmH2O, pulmonary arterial pressure = 10 cmH2O, left atrial pressure = 0 cmH2O; flow = 0.11 +/- 0.06 ml.100 g body wt-1 x min-1) and rapidly froze them. Histologically, 33 +/- 19% (SD) of alveolar septa fluoresced, demonstrating that the perfusate had not been excluded. However, we could not resolve whether the fluorescence originated in the septal microvascular lumen or in the adjacent perimicrovascular interstitial space. To address this issue, we perfused an additional lung with horseradish peroxidase (HRP) and examined it by transmission electron microscopy. HRP filled interstitial spaces around septal vessels and extraseptal alveolar corner vessels, but because the septal vascular lumina were too compressed, we were unable to determine whether they also contained HRP. Therefore we perfused two additional lungs with particles of colloidal gold (0.05 microns diam). Using transmission electron microscopy, we found gold particles in 15-25% of septal vascular lumina, demonstrating that septal vessels were at least partially accessible in zone 1. Our interpretations is that filtration in zone 1 may occur from septal vessels and extraseptal alveolar vessels. Furthermore, results of the HRP study suggest that the perimicrovascular interstitial space is less compressible than the septal vascular lumen. PMID- 8226473 TI - Human muscle metabolism during intermittent maximal exercise. AB - Eight male subjects volunteered to take part in this study. The exercise protocol consisted of ten 6-s maximal sprints with 30 s of recovery between each sprint on a cycle ergometer. Needle biopsy samples were taken from the vastus lateralis muscle before and after the first sprint and 10 s before and immediately after the tenth sprint. The energy required to sustain the high mean power output (MPO) that was generated over the first 6-s sprint (870.0 +/- 159.2 W) was provided by an equal contribution from phosphocreatine (PCr) degradation and anaerobic glycolysis. Indeed, within the first 6-s bout of maximal exercise PCr concentration had fallen by 57% and muscle lactate concentration had increased to 28.6 mmol/kg dry wt, confirming significant glycolytic activity. However, in the tenth sprint there was no change in muscle lactate concentration even though MPO was reduced only to 73% of that generated in the first sprint. This reduced glycogenolysis occurred despite the high plasma epinephrine concentration of 5.1 +/- 1.5 nmol/l after sprint 9. In face of a considerable reduction in the contribution of anaerobic glycogenolysis to ATP production, it was suggested that, during the last sprint, power output was supported by energy that was mainly derived from PCr degradation and an increased aerobic metabolism. PMID- 8226474 TI - Changes in resistance and ventilatory timing that accompany apnea in premature infants. AB - To characterize the changes in respiratory mechanics and ventilatory timing that accompany apnea in premature infants, we evaluated 36 apneas in 13 premature infants (birth weight, 1,200 +/- 350 g, postconceptional age at study 34 +/- 3 wk). Apnea was defined as a ventilatory pause > or = 10s accompanied by a decrease in heart rate of 20 beats/min. Nasal airflow was recorded with a pneumotachometer, and esophageal, pharyngeal, and nasal mask pressures were continuously measured. Inspiratory time (TI), expiratory time (TE), tidal volume (VT), and VT/TI were determined over five breaths before and after apnea. In addition, total pulmonary resistance (RT) and supraglottic resistance (Rs) were measured over the same epochs in inspiration and expiration. Before apnea, TE and RT increased (P < 0.05 and < 0.01, respectively); however, Rs did not change. Immediately after apnea, prolongation of TI occurred and both RT and Rs were increased (P < 0.01), consistent with continued upper airway instability. However, within two breaths after resolution of the apnea, RT and Rs returned to normal, reflecting rapid recovery of upper airway and total pulmonary resistance. The ventilatory changes that precede and follow apnea closely resemble those occurring during periodic breathing. PMID- 8226475 TI - Relationship between arterial and portal vein immunoreactive glucagon during exercise. AB - The importance of changes in glucagon in the regulation of hepatic glucose production (Ra) during exercise has been questioned, as an increase in arterial immunoreactive glucagon (IRG) is not always detectable. However, IRG in the portal vein (PV) and not in the artery is most relevant, as flow through PV is approximately 80% of liver blood flow. To assess the extent that arterial IRG reflects the levels the liver is exposed to in PV, dogs (n = 5) were implanted with catheters in a carotid artery, hepatic vein (HV), and PV. Dogs were studied > or = 16 days later during rest and 150 min of moderate treadmill exercise, with indocyanine green and [3-3H]glucose infused to assess hepatic plasma flow (HPF) and hepatic Ra. IRG was 66 +/- 7, 73 +/- 8, and 81 +/- 7 pg/ml in the artery, HV, and PV at rest; it rose at 10 and 150 min of exercise to 89 +/- 9 and 127 +/- 13 pg/ml in the artery, 106 +/- 17 and 186 +/- 21 pg/ml in HV, and, by considerably more, to 153 +/- 20 and 261 +/- 25 pg/ml in PV. HPF fell by approximately 30% with exercise. The fall in HPF accounted for < 11% of the increased arterial-to PV IRG gradient during exercise, with increased splanchnic IRG release comprising the remainder. Ra was linearly related to IRG levels in the three vessels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226476 TI - Mathematical model of cycling performance. AB - A model of cycling performance is presented. The model is based on equating two expressions for the total amount of work performed. One expression is deduced from biomechanical principles deriving energy requirements from total resistance. The other models the energy available from aerobic and anaerobic energy systems, including the effect of oxygen uptake kinetics at the onset of exercise. The equation can then be solved for any of the variables. Empirically derived field and laboratory data were used to assess the accuracy of the model. Model estimates of 4,000-m individual pursuit performance times showed a correlation of 0.803 (P < or = 0.0001) with times measured in 18 high-performance track cyclists, with a mean difference (predicted--measured) of 4.6 s (1.3% of mean performance time). The model enables estimates of the performance impact of alterations in physiological, biomechanical, anthropometric, and environmental parameters. PMID- 8226477 TI - Limitation of airway smooth muscle shortening by cartilage stiffness and lung elastic recoil in rabbits. AB - Airway smooth muscle can contract to 20% of its starting length when stimulated maximally and allowed to contract isotonically in vitro. In vivo airway smooth muscle contraction of this degree would result in widespread airway closure. We hypothesized that elastic loads related to cartilage stiffness and lung parenchyma-airway interdependence limit in vivo airway smooth muscle shortening. We measured pulmonary resistance in anesthetized tracheostomized New Zealand White rabbits before and after intravenous treatment with papain in a concentration that produced generalized cartilage softening. Papain treatment caused a significant increase in pulmonary resistance that was completely reversed by application of 4 cmH2O positive end-expiratory pressure and that was partially reversed by vagotomy. Papain pretreatment also resulted in a substantial alteration in the pulmonary resistance-dose relationship to intravenously administered acetylcholine. In addition, maximal resistance after the highest concentration of acetylcholine was greater in papain-treated animals than in the control animals, but the position of the dose-response relationship was not shifted (i.e., there was no change in the effective dose causing 50% maximal response). Application of 4 cmH2O positive end-expiratory pressure in untreated animals resulted in a marked decrease in the bronchoconstriction produced by an effective dose of acetylcholine causing 50% of maximal response, whereas application of 4 cmH2O negative end-expiratory pressure resulted in a marked enhancement of the bronchoconstrictor response to the same intravenous dose of acetylcholine. We conclude that cartilage elasticity and lung recoil are important determinants of the ability of airway smooth muscle to shorten and produce airway narrowing in vivo. PMID- 8226478 TI - Respiratory gas exchange and metabolic responses during exercise in McArdle's disease. AB - During normal progressive exercise, the gas exchange anaerobic threshold occurs when CO2 production (VCO2) and ventilation (VE) increase so as to depart from a linear relationship to O2 consumption (VO2). This is thought to represent a gas exchange response to metabolic acidosis due to lactate accumulation. Patients with McArdle's disease have previously been reported to exhibit a steepened ventilatory response relative to VCO2, despite an inability to produce lactate. However, the VCO2 response has not been studied. We therefore investigated the VCO2-VO2 and VE-VO2 relationships in seven McArdle's disease patients and seven control subjects during symptom-limited maximal treadmill exercise. Analysis of gas exchange showed that whereas all control subjects had an easily identifiable anaerobic threshold, four of the patients had none and the other three displayed an attenuated threshold. The occurrence of the threshold in one patient was associated with a small rise in lactate and in another patient with an abrupt rise in leg discomfort, suggesting a pain response. Ammonia and the purine metabolite hypoxanthine were elevated during exercise in all patients, suggesting that ammonia may be a product of adenosine monophosphate degradation. Free fatty acid levels were also elevated, and a shift toward utilization of lipid may contribute to abnormal gas exchange responses. It is concluded that lactic acidosis contributes to the gas exchange anaerobic threshold but that other factors, such as discomfort, may be involved in the excess Ve seen during heavy exercise. PMID- 8226479 TI - O2 uptake kinetics and the O2 deficit as related to exercise intensity and blood lactate. AB - The dynamic responses of O2 uptake (VO2) to a range of constant power output levels were related to exercise intensity [as percent maximal VO2 and as below vs. above lactic acid threshold (LAT)] and to the associated end-exercise lactate in three groups of subjects: group I, untrained subjects performing leg cycle ergometer exercise; group II, the same subjects performing arm cycle exercise; and group III, trained cyclists performing leg cycle ergometer exercise. Responses were described by a double-exponential equation, with each component having an independent time delay, which reduced to a monoexponential description for moderate (below-LAT) exercise. When a second exponential component to the VO2 response was present, it did not become evident until approximately 80-100 s into exercise. An overall time constant (tau T, determined as O2 deficit for the total response divided by net end-exercise VO2) and a primary time constant (tau P, determined from the O2 deficit and the amplitude for the early primary VO2 response) were compared. The tau T rose with power output and end-exercise lactate levels, but tau P was virtually invariant, even at high end-exercise lactate levels. Moreover the gain of the primary exponential component (as delta VO2/delta W) was constant across power outputs and blood lactate levels, suggesting that the primary VO2 response reflects a linear system, even at higher power outputs. These results suggest that elevated end-exercise lactate is not associated with any discernible slowing of the primary rise in VO2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226480 TI - Exercise and glucocorticoid-induced diaphragmatic myopathy. AB - We tested the hypothesis that a chronically active muscle, such as the rat diaphragm, would be more resistant to glucocorticoid-induced myopathy than a less active locomotor skeletal muscle (plantaris). Furthermore, we sought to determine whether endurance exercise could antagonize the glucocorticoid-induced atrophy in the diaphragm. Rats were assigned to one of seven experimental groups (n = 10 per group) and injected daily over a 10-day period with either a sham solution or prednisolone acetate: group 1: control; sedentary and sham injected; group 2: control; exercise trained and sham injected; group 3; sedentary; prednisolone (0.5 mg.kg-1 x day-1); group 4: sedentary; prednisolone (1.0 mg.kg-1 x day-1); group 5: sedentary; prednisolone (2.0 mg.kg-1 x day-1); group 6: sedentary; prednisolone (5.0 mg.kg-1 x day-1); group 7: exercise trained; prednisolone (5.0 mg.kg-1 x day-1). Slope differences in the dose-response curves suggest that prednisolone-induced muscle atrophy in the plantaris was more severe than that in the diaphragm. Furthermore, high doses of prednisolone resulted in a differential effect on muscle bioenergetic enzyme activities in the plantaris and diaphragm. Prednisolone treatment (> or = 2 mg.kg-1 x day-1) resulted in a significant reduction in phosphofructokinase activity (expressed as microM substrate.min-1 x mg protein-1) and an increase in 3-hydroxyacyl-CoA dehydrogenase activity in the plantaris muscle. In contrast, prednisolone treatment did not influence phosphofructokinase activity (P > 0.05) in the diaphragm but decreased (P < 0.05) relative citrate synthase activity. Finally, 90 min daily of endurance exercise did not antagonize prednisolone-induced myopathy in either the diaphragm or the plantaris. PMID- 8226481 TI - Effects of high-frequency ventilation and PEEP on carotid baroreceptor reflexes. AB - We tested the hypothesis that altering the pattern and/or magnitude of discharge of pulmonary stretch receptors (PSRs) would alter baroreceptor reflexes in anesthetized aortic-denervated cats. Carotid baroreceptor control of mean arterial pressure (MAP), heart rate (HR), and hindlimb perfusion pressure (PPhl) was examined by changing carotid sinus pressure (CSP) from 50 to 225 mmHg. The pattern of PSR discharge was changed by switching conventional mechanical ventilation (CMV) to high-frequency ventilation (HFV). Magnitude of PSR discharge was altered by changing positive end-expiratory pressure (PEEP). Altering the discharge pattern of PSR had no effect on CSP-MAP or CSP-PPhl relationships; small changes in HR were observed. Increasing PSR activity by increasing PEEP during CMV (from 3 to 9 cmH2O) depressed CSP-MAP relationship, set point, and threshold pressure. However, the depression in CSP-MAP relationship and set point during PEEP was unrelated to PSR activation, because these changes were not abolished after bilateral vagotomy. CSP-PPhl relationship was significantly elevated during PEEP before and after vagotomy, suggesting activation of a nonvagally mediated vasoconstrictory mechanism instead of PSR-mediated depressor reflex. CSP-HR relationship during PEEP showed a slight elevation, which was abolished after vagotomy. We conclude that despite minor increases in HR, altering the pattern of magnitude of PSR activity with HFV and PEEP has no significant effect on carotid baroreceptor regulation of systemic circulation. Hemodynamic changes observed during PEEP were likely due to its mechanical effect on cardiac output and activation of other cardiopulmonary receptors rather than to the increase in PSR activity. PMID- 8226482 TI - On the preservation and regulation of vascular tone in arteriovenous anastomoses during anesthesia. AB - In conscious pigs, arteriovenous anastomoses (AVAs) are in a constricted state so that < 5% of intra-atrially injected radioactive (15-microns-diam) microspheres are shunted to the lungs. Many of the anesthetic regimens frequently used in cardiovascular research dilate AVAs, thereby greatly increasing the percentage of microspheres reaching the lungs. This may seriously limit extrapolation of results obtained under anesthesia to the conscious state. We now describe that anesthesia with a combination of fentanyl and thiopental preserves the tone of AVAs, maintaining shunting under 4% of cardiac output. Furthermore, we studied in the carotid circulation of this model whether norepinephrine or 5-hydroxy tryptamine (5-HT), both contained in perivascular nerves, is responsible for this tone. Consecutive antagonism of alpha 1-, alpha 2-, 5-HT1, and 5-HT2 receptors was obtained by sequential injection of prazosin, phentolamine, ketanserin, and methiothepin. Prazosin increased AVA blood flow, partly at the expense of extracerebral tissue blood flow, but preserved cerebral blood flow. None of the other antagonists had any additional significant effect. Therefore, in this model the tone in AVAs seems to be maintained by sympathetic norepinephrine-containing nerves via alpha 1-adrenoceptors. PMID- 8226483 TI - Effects of muscle contraction on cytochrome a,a3 redox state. AB - The relationships among mitochondrial O2 availability, O2 delivery, and lactate formation in exercising skeletal muscle remain unclear. Some data suggest that muscle O2 provision is sufficient at maximal O2 consumption (VO2max) to challenge the concept of a mitochondrial O2 limitation at VO2max. The relationships among VO2, mitochondrial O2 availability, and net lactate production were studied over a wide range of exercise intensities. Using near-infrared spectroscopy, the oxidation-reduction state of cytochrome a,a3 was monitored in the canine gracilis in vivo. Twenty adult dogs were anesthetized with alpha-chloralose, intubated, and mechanically ventilated on room air. Five-minute stimulation periods at rates of 2, 3, 4, 5, 7, 8, 10, or 12 stimuli/s were performed. VO2max generally was achieved at a stimulation rate of 8 stimuli/s; mean VO2max was 0.12 +/- 0.09 (SE) ml.min-1 x g-1. The concentration of oxidized mitochondrial cytochrome a,a3 decreased at all work loads relative to resting state and demonstrated a near linear relationship with muscle VO2 (r2 = 0.99). Muscle lactate efflux and the lactate-pyruvate ratio also were correlated positively with cytochrome a,a3 reduction, suggesting a common regulatory mechanism coupling the processes of aerobic glycolysis and oxidative phosphorylation. At VO2max, the corresponding cytochrome oxidation was not significantly different from that observed at death. Thus, in the gracilis maximal exercise leads to near-complete reduction of cytochrome a,a3 secondary to deficient O2 provision. We conclude that VO2max is limited primarily by O2 delivery to this muscle and not by other factors limiting mitochondrial ATP production or substrate oxidation. PMID- 8226484 TI - Effects of capsaicin desensitization on nasal mucosal secretion in guinea pigs in vivo. AB - Capsaicin-sensitive mechanisms may contribute to histamine's effects on guinea pig nasal mucosal secretion in vivo. Histamine nasal provocations were performed, secretions were collected, and the following variables were measured: total protein (marker of all secretory processes), alkaline phosphatase (marker of glandular secretion), and albumin (marker of vascular permeability, epithelial permeability, and glandular transport). By challenging only one nostril (ipsilateral), the contralateral responses to these challenges were examined and the nature of "nasonasal" reflexes was determined. Histamine increased albumin and alkaline phosphatase concentrations in lavage fluid on the ipsilateral and contralateral sides. The alkaline phosphatase secretion was reduced by atropine. Capsaicin induced dose-dependent albumin exudation. To determine the roles of capsaicin-sensitive sensory nerves, one nostril was desensitized by topical application of capsaicin to one nostril in increasing doses daily for 5 wk. This led to 1) ablation of all secretory responses to capsaicin, 2) ablation of histamine-induced alkaline phosphatase and total protein secretion, 3) blunted histamine-induced albumin exudation on the desensitized side, and 4) ablation of histamine-induced contralateral reflex-mediated protein secretion. These results indicate that capsaicin stimulates vascular responses, probably by stimulating nociceptive nerve axon responses. Histamine induces vascular responses and albumin exudation by both capsaicin-sensitive sensory neuron axon responses and direct effects on vessels. Histamine stimulates glandular secretion through a capsaicin-sensitive afferent/cholinergic efferent reflex. PMID- 8226485 TI - Caffeine metabolism and epinephrine responses during exercise in users and nonusers. AB - This study compared the caffeine (CAF) metabolism and the catecholamine and metabolic responses of users and nonusers of caffeine after acute ingestion of caffeine (5 mg/kg) during 1 h of steady-state exercise (50% maximal oxygen consumption). Nonusers (n = 7) completed two exercise trials after ingesting either CAF (5 mg/kg) or placebo (PL). Users (n = 7) underwent three trials designed to control caffeine use and abstained from voluntary CAF intake for 18 days. After 4 days they had a PL trial and in the following 14 days they were given random 6 days of CAF (2 x 2.5 mg.kg-1 x day-1) or PL ingestion followed in each case on the 7th day by a CAF exercise trial identical to that of the nonusers. In nonusers CAF increased (P < 0.05) plasma epinephrine (EPI) concentration above PL values during exercise. Users did not exhibit any increased EPI with CAF, but the EPI response to exercise in all three trials was twofold greater than that of the nonusers' PL trial (P < 0.05). In all trials both groups had identical norepinephrine responses. The groups had similar plasma and urinary caffeine concentration, but plasma dimethylxanthines varied; the users had greater (P < 0.05) theophylline concentration, and the nonusers had a greater (P < 0.05) rise in paraxanthine (PX) concentration. The users and nonusers' plasma free fatty acids (FFA), glycerol and respiratory exchange ratio were similar after ingestion of CAF. Although PX may increase FFA in resting subjects, in this study PX concentrations in nonusers varied from that of the users, yet FFA data were similar.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226486 TI - Relationships between in vivo and in vitro measurements of metabolism in young and old human calf muscles. AB - This study compared in vivo measurements of muscle metabolism in humans with magnetic resonance spectroscopy (MRS) and in vitro analysis of biopsies. Healthy subjects [4 young males, 28.2 +/- 6.8 (SD) yr, and 6 older subjects (5 males, 1 female), 66 +/- 6.0 yr] performed a maximal cycle ergometer test, and MRS measurements of the calf muscles and needle biopsies of the lateral gastrocnemius were performed. Biopsies were analyzed for fiber type and citrate synthase (CS) activity. MRS measurements of inorganic phosphate (Pi), phosphocreatine (PCr), ATP, and pH were made using a 1.8-T 78-cm clear-bore magnet-and-spectrometer system. Two or three 5-min bouts of plantar flexion were performed against variable resistance to deplete PCr levels to 50% of resting values (mean end pH 6.99). PCr values during recovery were fit to an exponential curve, and the rate constant (PCrrate) was calculated. PCrrate was used as an index of oxidative metabolism. Older subjects had lower peak O2 uptake (VO2 peak) values (19.2 +/- 5.6 vs. 49.5 +/- 8.1 ml O2.min-1 x kg-1), CS activities (16 +/- 2.8 vs. 25 +/- 2.6 mmol.kg wet wt-1 x min-1), and PCrrate values (25.3 +/- 8. vs. 37.5 +/- 5.3 mmol PCr.kg wet wt-1.min-1) than young subjects. PCrrate correlated with CS activity, and both PCrrate and CS activity correlated with VO2 peak (P < 0.05). No correlations were found between percent fiber type and PCrrate, CS activity, and VO2 peak. These results support studies that showed decreases in muscle metabolism with age in healthy humans and show a good correlation between in vivo and in vitro measurements of oxidative metabolism. PMID- 8226487 TI - Effect of insulin-like factors on glucose transport activity in unweighted rat skeletal muscle. AB - We have previously demonstrated that mechanical unweighting of the soleus muscle by hindlimb suspension leads to increases in insulin receptor binding, muscle/fat specific glucose transporter (GLUT-4) protein levels, and insulin-stimulated glucose transport activity. The present study used a novel approach to further evaluate the potential role of postreceptor binding mechanisms in this enhanced insulin effect after unweighting. Insulin-like growth factor I (IGF-I), vanadate, and phospholipase C were used to stimulate glucose transport activity independently of insulin receptor binding. Soleus glucose transport activity (assessed by 2-deoxyglucose uptake) was evaluated in vitro with soleus strips (approximately 18 mg). Progressively increased responses to maximally effective doses of insulin or IGF-I were observed after 3 and 6 days of unweighting compared with weight-matched control strips. Enhanced maximal responses to vanadate (6 days only) and phospholipase C (3 and 6 days) for 2-deoxyglucose uptake were also observed. The results of this study 1) provide evidence that post-insulin receptor binding mechanisms also play a role in the enhanced response of the insulin-dependent pathway for stimulation of glucose transport in unweighted skeletal muscle and 2) indicate that IGF-I action on glucose transport is included in this enhanced response in unweighted muscle. PMID- 8226488 TI - Heat acclimation improves cardiac mechanics and metabolic performance during ischemia and reperfusion. AB - Cardiac mechanics and metabolic performance were studied in isolated perfused hearts of heat-acclimated (AC) rats (at 34 degrees C for 1 mo) and their age matched controls (C). Diastolic and systolic pressures, coronary flow, and the appearance of ischemic contracture (IC) were measured during progressive graded ischemia, total ischemia (TI), and reperfusion. ATP, phosphocreatine, and intracellular pH were measured during TI and reperfusion with the use of 31P nuclear magnetic resonance spectroscopy. Systolic pressure was greater in AC hearts than in C hearts (P < 0.0001). During 50% of perfusion pressure 15 and 46% of AC and C hearts, respectively, showed IC (P < 0.001). During 25% of perfusion pressure 85% of the hearts in both groups developed IC. The onset of IC in AC hearts was delayed compared with in C hearts. On reperfusion 93 and 66% of AC and C hearts, respectively, resumed contraction. Recovery of diastolic pressure was 78 and 36% for the AC and C hearts, respectively (P < 0.05). During TI ATP declined by 0.94 and 1.20 mumol/min in AC and C hearts, respectively, resulting in 21 +/- 2.8% preservation of the ATP pool in AC hearts after 30 min of TI (P < 0.001). The AC group also showed a delayed decline in intracellular pH (P < 0.001). The data suggest beneficial effects of heat acclimation on the heart, which were exhibited by greater pressure generation and by the emergence of protecting features during ischemia and reperfusion, possibly via energy-sparing mechanisms. PMID- 8226489 TI - Pulmonary arterial elasticity in awake dogs. AB - We measured the relationship between pulmonary arterial pressure (Ppa), diameter (D), and length of a segment of the main pulmonary artery (MPA) in chronically instrumented conscious dogs breathing spontaneously (CCC). There were no physiologically significant changes in Ppa or D in the CCC dogs postoperatively, and the cross-sectional MPA shape measured by fast computed tomography was nearly circular. These results suggest that the MPA was not distorted by chronic instrumentation. We compared measurements made in the CCC dogs with previous measurements in acutely instrumented anesthetized dogs with open chests (AAO). The elasticity of MPA in the CCC animals was frequency dependent between 1 and 14 Hz and was similar to that in the AAO dogs. Oscillations of D preceded Ppa at cardiac frequencies in the AAO animals, but the D and Ppa oscillations were in phase in the CCC animals. The oscillations of length relative to D were significantly less in the CCC than in the AAO dogs. We conclude that, with limitations, the hemodynamic properties of the MPA can be measured in the CCC subjects. We suggest that the discrepancies between the AAO and CCC dogs can be attributed to differences in extrinsic loading of the MPA. PMID- 8226490 TI - Differences in cardiovascular adaptations to endurance exercise training between older men and women. AB - Recent studies have shown that people in their seventh decade are able to adapt to exercise training with an increase in maximal O2 uptake (VO2max) similar in relative magnitude to that observed in young people. The purpose of this study was to determine the relative contributions of increases in stroke volume and O2 extraction to the training-induced increase in VO2max in older men and women. Fifteen men [63 +/- 3 (SE) yr] and 16 women (64 +/- 3 yr), in good health, participated in 9-12 mo of endurance exercise training at 70 to 85% of maximal heart rate for 45 min/day, 4 days/wk. VO2max increased 19% (2.35 +/- 0.1 to 2.8 +/- 0.1 l/min; P < 0.01) in the men and 22% (1.36 +/- 0.1 to 1.66 +/- 0.1 l/min; P < 0.01) in the women in response to training. In the men, stroke volume during maximal exercise was 15% higher after training, and this increase accounted for 66% of the increase in VO2max. The remainder of the increase in VO2max was accounted for by a 7% greater arteriovenous O2 content difference during maximal exercise. In contrast, training resulted in no change in stroke volume in women, in whom the entire increase in VO2max was accounted for by a greater arteriovenous O2 content difference (12.2 +/- 0.4 before vs. 14.4 +/- 0.4 ml O2/100 ml blood after; P < 0.01) during maximal exercise. There were no changes in these variables in the control subjects. The mechanisms responsible for the training-induced increase in VO2max in the older men were similar to those found in young people.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226491 TI - Effects of high-frequency oscillating pressures on upper airway muscles in humans. AB - We examined the effects of high-frequency- (30-Hz) low-pressure oscillations (< 1 cmH2O) applied to the upper airway, via a nose mask, on genioglossus (EMGgg), sternomastoid (EMGsm), and diaphragm electromyogram (EMGdia) activity in sleeping humans. Ten patients with sleep apnea and six normal subjects were studied. The pressure oscillations were applied through the mask for a single breath. The subjects were studied in non-rapid-eye-movement (NREM) and rapid-eye-movement (REM) sleep. In the normal subjects, during NREM sleep, peak EMGgg, EMGsm, and EMGdia activity increased significantly in response to the oscillations in 63, 51, and 46%, respectively, of all trials. During REM sleep, significant increases occurred in 73, 88, and 13%, respectively, of all trials. Similar responses were observed in the patients with obstructive sleep apnea. Peak EMGgg, EMGsm, and EMGdia activity increased significantly in 74, 50, and 67%, respectively, of all NREM sleep trials and in 55, 81, and 76%, respectively, of all REM sleep trials. An important finding was that in 46% of the trials in the patients with sleep apnea the oscillation-induced increase in EMGgg activity was associated with a partial or complete reversal of the upper airway obstruction with an increase in tidal volume. This was observed in NREM and REM sleep. We conclude that there are upper airway receptors that respond to low-pressure-high-frequency oscillations applied to the upper airway that have input to the genioglossus and other muscles of respiration. These responses may be utilized in future treatment for sleep apnea. PMID- 8226492 TI - Exercise training alters cardiovascular and hormonal responses to intracranial self-stimulation. AB - Both reinforcing intracranial self-stimulation (ICSS) and physical exercise result in heightened cardiovascular and endocrine responses. This study compared the cardiovascular and endocrine responses to ICSS in rats after either chronic ICSS or treadmill running. Male rats (n = 35) were implanted with bipolar electrodes aimed at the ventral tegmental area of the brain, and those that performed vigorous lever pressing for ICSS (> 50 presses/min; n = 30) were counter balanced into three groups: chronic ICSS (PRESS), chronic run training (RUN), or sedentary controls (CONT). PRESS, RUN, and CONT rats performed ICSS, ran on a motorized treadmill, or sat quietly in cages for 30 min/day, 5 day/wk, for 12 wk, respectively. All animals then performed 30 min of lever pressing for ICSS and were immediately killed. Oxygen consumption, heart rate, blood pressure, rectal temperature, and plasma norepinephrine, epinephrine, and corticosterone increased (P < 0.05) for all groups during lever pressing. PRESS rats did not differ from CONT rats for any variable studied. However, heart rate was lower and oxygen consumption, norepinephrine, and corticosterone were higher in RUN than in CONT rats. Heart and ventricle weights were higher in PRESS and RUN than in CONT rats; body weights were not different. These data suggest that chronic treadmill running results in adaptations that influence cardiovascular and hormonal responses to ICSS. PMID- 8226493 TI - Cholinergic and opioid involvement in release of growth hormone during exercise and recovery. AB - Cholinergic and opioid pathways have been implicated as mediators of the increased growth hormone (GH) release observed during exercise. This study compared the GH responses induced by a moderate-intensity exercise bout during treatment with placebo (Plac), the opioid receptor antagonist naltrexone (Nalt), the indirect cholinergic agonist pyridostigmine (PD), or a combination of the two drugs (P + N). Ten active males served as subjects (age, 25.1 +/- 0.6 yr; wt, 79.7 +/- 2.5 kg; % body fat, 14.9 +/- 1.4; peak oxygen consumption, 46.2 +/- 2.7 ml.kg-1 x min-1). Blood samples were drawn at 5-min intervals during the 4.5-h testing period to determine the GH concentration. The testing period was divided as follows: 0600-700 h = baseline, 0700-0800 h = preexercise, 0800-0830 h = exercise, and 0830-1030 h = recovery. Drugs were administered 1 h before exercise (at 0700 h). Exercise consisted of 30 min of cycling at an individualized work load previously found to elicit a blood lactate concentration of 2.5 mM. Heart rate, oxygen consumption, blood lactate, and blood glucose were measured throughout the exercise period. Results indicated that neither the resting GH concentration nor the metabolic parameters during exercise were altered by the treatments. Peak serum GH concentration was not significantly altered by the treatments (range 7.3 +/- 2.0 to 12.6 +/- 4.4 micrograms/l).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226494 TI - Exercise increases inflammatory macrophage antitumor cytotoxicity. AB - This study determined the effects of exercise on the ability of inflammatory macrophages to inhibit tumor cell growth in vitro (macrophage cytotoxicity). Thioglycollate injection (1 ml ip) was used as an inflammatory challenge and to partially activate macrophages for cytotoxicity. Inbred male C3H/HeN mice (n = 180) exercised moderately (MOD, 18 m/min, 30 min/day, 5% grade) or to exhaustion (EXH, 18-35 m/min, 2-4 h, 5% grade) on a motor-driven treadmill for 3 consecutive days after injection. Control (CON) mice were kept in stimulated treadmill lanes directly over the runners. Mice were killed immediately or 3 or 8 h postexercise. Macrophages from both MOD and EXH exercise groups manifested significantly (P < 0.05) enhanced (approximately 50%) cytotoxicity compared with those from CON group at all time points postexercise. This potentially beneficial exercise effect was not related to macrophage production of interleukin-1 beta, reactive nitrogen or oxygen intermediates, or number of macrophages in the assay but may have been manifested, in part, by tumor necrosis factor-alpha. Plasma corticosterone was significantly elevated immediately postexercise in MOD and EXH compared with CON mice; however, no evidence existed for an immuno-suppressive effect of corticosterone on macrophage cytotoxicity, perhaps because of insensitivity of inflammatory macrophages to glucocorticoid suppression seen in vitro. These data only partially support the "inverted U hypothesis," which states that moderate exercise may enhance, whereas very heavy exercise or a lack of exercise may attenuate, the immune response. Further study is needed to determine the physiological significance of these findings and the effects of exercise on macrophage subsets sensitive to glucocorticoid suppression (i.e., fully activated macrophages). PMID- 8226495 TI - Bifurcations of the respiratory pattern associated with reduced lung volume in the rat. AB - Rats with intact vagal reflexes exhibit patterns of breathing that contain greater degrees of freedom than those seen after vagotomy. To determine how alterations in end-expiratory volume modify the respiratory pattern, continuous positive (CPAP) and negative (CNAP) airway pressure was applied to tracheal openings of nine urethan-anesthetized vagi-intact rats (+3 to -9 cmH2O). Phase portraits (e.g., volume vs. flow curves), power spectra, correlation integral curves, and inspiratory-to-expiratory duration (TI/TE) ratios are used to interpret the vagal-dependent responses to changes in mean tracheal pressure (Ptr). With CPAP, respiratory oscillation was highly periodic and one dimensional, with TI/TE near 1.0. As Ptr was reduced in a stepwise manner, transient bursts of inspiratory airflow developed local to the expiratory inspiratory transition, with amplitude increasing proportionally with the level of CNAP. These oscillatory "expiratory interrupts" (doubling TI/TE in five of nine cases) produced highly variable and asymmetric respiratory patterns. Progressive increases in correlation dimension (maximum = 1.8-3.0) and tendencies toward broadband power spectra were seen as Ptr was lowered. The irregular phase switching dynamics seen with CNAP (which disappeared after vagotomy) are consistent with onset of low-dimensional chaos, probably correlated with activation of feedback mechanisms responsive to lung deflation. PMID- 8226496 TI - Role of deflation-sensitive feedback in control of end-expiratory volume in rats. AB - Rats breathing from reduced end-expiratory volumes (EEV) exhibit transient bursting of inspiratory airflow local to the expiratory-inspiratory transition (E I) accompanied by increases in inspiration-to-expiration duration ratio. Continuous positive (CPAP) and negative (CNAP) airway pressures (+3 to -9 cmH2O) were applied to tracheal openings of 17 unilaterally vagotomized rats (urethan anesthetized) to evaluate the feedback mechanisms associated with these oscillatory "expiratory interrupts." Whole nerve recordings of afferent vagus (Vag) were primarily inspiratory with CPAP. As tracheal pressure was reduced in a stepwise manner, progressive decreases in inspiratory peaks and increases in activity during late expiration were seen on Vag. Single-fiber recordings correlate Vag expiratory activity with slowly adapting receptors sensitive to lung deflation. With CNAP, 1) progressively earlier onsets of activity in inspiratory muscles (e.g., diaphragm, genioglossus, posterior cricoarytenoid) were observed (increased inspiratory duration, decreased expiratory duration); oscillations at E-I developed only when expiratory peaks on Vag were significant. 2) Thyroarytenoid muscle exhibited onset of electromyographic activity that immediately succeeded onset of Vag during late expiration. When the trachea is intact, these two actions might combine to produce a respiratory pattern similar to the "grunting" often seen in newborns breathing from reduced EEVs. In contrast to larger mammals, rats rely strongly on deflation reflexes for control of E-I phase switching, presumably to aid in maintenance of an elevated EEV. PMID- 8226497 TI - Bifurcations of the respiratory pattern produced with phasic vagal stimulation in the rat. AB - Geometric methods from nonlinear dynamics are employed to evaluate dynamic processing of vagal afferent information by the respiratory central pattern generator (RCPG). While measuring airflow and diaphragm EMG, we applied brief electrical stimuli (40- to 130-ms duration) to one afferent vagus of bilaterally vagotomized urethan-anesthetized rats during every breath at various phases of the respiratory cycle. Stimuli applied during early or late inspiration of every breath evoke highly predictable one-dimensional responses: reversible (graded inhibition) or irreversible (off-switching) inhibition of inspiratory activity, respectively. Stimulation during midinspiration produces higher-dimensional oscillations that wander unpredictably over a continuum of graded inhibition and off-switching; "spiral" attractors and "horseshoe" return maps at this phase are characteristic of Silnikov's bifurcation. Stimuli applied during early expiration always prolonged expiratory duration, but those delivered during midexpiration evoked unpredictable wandering between prolongations and shortening of expiratory duration. A narrow time window surrounds the expiratory-inspiratory (E-I) transition, where stimuli elicit either breaths of short duration and low amplitude (irreversible E-I transition, decreased total respiratory cycle duration) or transient bursts of inspiratory activity at the E-I transition followed by a prolonged breath (reversible E-I transition, increased total respiratory cycle duration). We conclude that RCPG "gating" of and adaptation to vagal feedback combine to produce complex breath-to-breath dynamics in the rat that are consistent with low-dimensional chaos. PMID- 8226498 TI - Effect of ventilation inhomogeneity on "intrabreath" measurements of diffusing capacity in normal subjects. AB - In normal seated subjects we increased single-breath ventilation inhomogeneity by changing both the preinspiratory lung volume and breath-hold time and examined the ensuing effects on two different techniques of measuring the diffusing capacity of the lung for carbon monoxide (DLCO). We measured the mean single breath DLCO using the three-equation method (DLCOSB-3EQ) and also measured DLCO over discrete intervals during exhalation by the "intrabreath" method (DLCOexhaled). We assessed the distribution of ventilation using the normalized phase III slope for helium (SN). DLCOSB-3EQ was unaffected by preinspiratory lung volume and breath-hold time. DLCOexhaled increased with increasing preinspiratory lung volume and decreased with increasing breath-hold time. These changes correlated with the simultaneously observed changes in ventilation inhomogeneity as measured by SN (P < 0.01). We conclude that measurements of DLCOexhaled do not accurately reflect the mean DLCO. Intrabreath methods of measuring DLCO are based on the slope of the exhaled CO concentration curve, which is affected by both ventilation and diffusion inhomogeneities. Although DLCOexhaled may theoretically provide information about the distribution of CO uptake, the concomitant effects of ventilation nonuniformity on DLCOexhaled may mimic or mask the effects of diffusion nonuniformity. PMID- 8226499 TI - Effect of temperature on alveolar liquid and protein clearance in an in situ perfused goat lung. AB - To study the mechanisms of alveolar liquid and protein clearance, we investigated the effect of four different temperatures (7, 18, 30, and 38 degrees C) on the rate of liquid, [14C]mannitol, and 125I-labeled albumin (125I-albumin) transport from the air spaces of isolated perfused in situ goat lungs over 4 h under isogravimetric conditions. The concentrations of both native proteins and 125I albumin in the alveolar fluid after 4 h were used to estimate alveolar liquid clearance. We instilled autologous serum (2 ml/kg) with 125I-albumin and [14C]mannitol into the distal air spaces of one lung and 131I-labeled albumin into the perfusate to measure the capillary-to-alveolar permeability to protein. Alveolar liquid clearance progressively declined from a control value of 24 +/- 6% at 38 degrees C to 12 +/- 7% at 30 degrees C, to 3 +/- 3% at 18 degrees C, and to -15 +/- 11% at 7 degrees C. 125I-albumin clearance into the perfusate was 0.3%/h of the instilled 125I-albumin at 38 degrees C. The clearance decreased abruptly to 0.06%/h at 30 degrees C and then slightly increased to 0.08%/h at 18 degrees C and to 0.15%/h at 7 degrees C. There were no changes in transport except for a twofold increase in mannitol at 7 degrees C. Increased permeability of the alveolar barrier to protein and small-molecular-weight tracers at 7 degrees C indicates that the barrier function of the alveolar epithelium is disrupted at this low temperature.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226500 TI - Hemodynamic effects of brief pulmonary hemostasis. AB - We determined the hemodynamic effects of brief hemostasis in isolated blood perfused rat lungs held at constant inflation pressure. Three periods of hemostasis, each lasting 20 min, alternated with equal durations of perfusion. Pulmonary vascular resistance (PVR), calculated as the ratio of the pulmonary arterial- (Ppa) left atrial (Pla) pressure difference to flow, was determined at baseline and after the third hemostasis period. Hemostasis increased PVR by 127% above baseline at constant Ppa and Pla (n = 8; P < 0.05) and by 71% at constant Pla and constant flow (n = 16; P < 0.05). The PVR increases were significantly attenuated by platelet (n = 5) or red blood cell (n = 4) removal from the lung perfusion, mechanical ventilation (n = 5), indomethacin (2 X 10(-6) M; n = 5), papaverine (10(-5) M; n = 4), and the thromboxane blocker SQ-29548 (7.8 X 10(-5) M; n = 4). By micropuncture we determined that the posthemostasis increase of the total pressure drop was 66% due to an increase of the venous pressure drop (P < 0.05). Isogravimetric pressure and lung water were not significantly altered by hemostasis. We conclude that brief hemostasis in the rat lung induces platelet- and red blood cell-dependent increase of vascular resistance. This effect may be attributable to hemostasis-induced platelet activation leading to thromboxane release. PMID- 8226501 TI - Metabolic response to halothane in piglets susceptible to malignant hyperthermia: an in vivo 31P-NMR study. AB - Using in vivo 31P-nuclear magnetic resonance spectroscopy, we studied the skeletal muscle metabolism of 17 anesthetized malignant hyperthermia-susceptible piglets and 25 control piglets during and after a halothane stress test. At rest, the phosphocreatine- (PCr) to-ATP ratio was 12% higher in the anesthetized piglets than in the control piglets, which may reflect a higher proportion of fast glycolytic fibers in the former. About 15 min of halothane administration sufficed to provoke onset of a reaction, which was characterized by a reciprocal drop in PCr and an increase in Pi with commencing intracellular acidosis. Halothane was withdrawn after a 20% drop in PCr. Within the next few minutes, intracellular pH dropped sharply and phosphomonoesters (PME) accumulated excessively. ATP was observed to decrease in 8 of the 17 animals. Halothane inhalation provoked a switch of metabolism toward glycolysis. Accumulation of PME suggests a mismatch between glycogenolysis and glycolysis. Despite severe acidification, glycolysis was not completely halted. Recovery of PCr and Pi started approximately 5 min after halothane withdrawal, with a longer time constant for recovery of the former. PME and intracellular pH aberrations lingered and started to recover later. Lost ATP was never restored within the observed recovery period of approximately 20 min. PMID- 8226502 TI - Imaging the ovine heparin-protamine interaction with 111In-protamine. AB - Protamine reversal of heparin anticoagulation occasionally induces the release of thromboxane into plasma with catastrophic pulmonary hypertension. To examine the site of neutralization, we labeled protamine sulfate with 111In and compared activity scans after administration of labeled protamine in unheparinized and heparin-anticoagulated sheep. Protamine administration in sheep without prior heparinization did not cause thromboxane release, pulmonary hypertension, or significant leukopenia, and 111In-protamine was rapidly cleared from the lungs (half time 0.48 +/- 0.08 min). Neutralization of heparin anticoagulation by labeled protamine produced elevated plasma thromboxane, pulmonary vasoconstriction, leukopenia, and prolonged pulmonary clearance of 111In protamine (half time 3.32 +/- 0.43 min). In rats, protamine reversal of heparin anticoagulation did not induce either thromboxane synthesis or pulmonary hypertension, and 111In-protamine cleared rapidly from the lungs. Thus the ovine heparin-protamine reaction produces concomitant pulmonary sequestration of heparin-protamine complexes, thromboxane release, and pulmonary vasoconstriction; this did not occur in the rat. The lung specificity of the reaction and interspecies differences suggest that ovine pulmonary intravascular macrophages may be activated by heparin-protamine complexes to release thromboxane and provoke acute pulmonary vasoconstriction. PMID- 8226503 TI - Persistent effects of exercise on skeletal muscle glucose transport across the life-span of rats. AB - Very young rats (< 2 mo) have a persistent increase in insulin-stimulated glucose transport rate in skeletal muscle for several hours after completing a bout of exercise. We studied the effect of exercise on the glucose transport activity of isolated epitrochlearis muscles from male Fischer 344/Brown Norway F1 hybrid rats across a wide range of the life-span (at 3.5, 13, and 25 mo). The stimulation of 3-O-methylglucose (3-MG) accumulation by a submaximally effective insulin concentration (100 microU/ml) was enhanced (50-75%) 4 h after exercise, regardless of age. In contrast, the 3-MG transport rate with 20,000 microU/ml insulin was enhanced after exercise only in the youngest rats (35%), and this increased responsiveness occurred despite no changes in muscle total GLUT-4 levels. In addition, epitrochlearis GLUT-4 levels were reduced by 29% between 3.5 and 13 mo of age in sedentary rats but did not decline further between 13 and 25 mo of age. GLUT-4 levels were moderately but significantly (P < 0.05) related (r = 0.554) to epitrochlearis muscle capacity for insulin-stimulated 3-MG transport. PMID- 8226504 TI - Forearm vascular responses to baroreceptor unloading at the onset of dynamic exercise. AB - To determine the extent to which reflexes accompanying muscular exercise (associated with central command) interact with cardiopulmonary (CP) baroreceptor mediated reflexes controlling forearm vascular resistance (FVR), we examined the forearm vasoconstrictor response at the onset of dynamic exercise, with and without CP baroreflex unloading, in 10 physically active men. CP baroreceptors were unloaded by application of lower body negative pressure (LBNP) at rest and during five 4-min bouts of supine exercise at 25 and 32 degrees C. Exercise intensities were 10 (essentially no load) and 100 W, and LBNP was applied at -10, -20, -30, and -40 mmHg during rest and at -20 and -40 mmHg during exercise. Resting FVR was 33.0 +/- 3.2 and 14.0 +/- 2.7 resistance units, and cardiac stroke volume (SV) was 117 +/- 7 and 126 +/- 9 ml/beat at 25 and 32 degrees C, respectively. We found a linear relationship between the increase in FVR and decrease in SV during LBNP; the slope of the relationship was significantly lower at 32 degrees C (FVR = 51.7-0.29SV) than at 25 degrees C (FVR = 123-0.79SV). At the onset of 100-W exercise without LBNP, FVR increased significantly to 50.2 +/- 9.0 and 21.2 +/- 3.2 units at 25 and 32 degrees C, respectively, whereas SV was unchanged. Application of -40-mmHg LBNP reduced SV significantly to 68 +/- 5 and 71 +/- 6 ml/beat and increased FVR significantly to 89.0 +/- 11.3 and 36.3 +/- 7.6 units at 25 and 32 degrees C, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226505 TI - Effects of air embolism on sheep lung fluid volumes. AB - Changes in lung fluid volumes and hyaluronan clearance were measured in six awake sheep during increased microvascular permeability induced by pulmonary air embolism (AE). After a 1- to 2-h baseline, filtered room air was infused through a proximal port of a Swan-Ganz catheter for 2 h at a rate sufficient to double pulmonary vascular resistance. The air infusion was discontinued, and the sheep were monitored for an additional 2 h (recovery). Lung lymph flow and protein flux increased during air infusion and continued to increase during recovery. During AE, lymph-to-plasma ratio for albumin decreased while lymph-to-plasma ratio for large protein remained the same. This would suggest that both microvascular pressure and microvascular permeability increase during AE. Protein clearance increased similarly for all protein sizes during AE and recovery. After 2 h of recovery, interstitial and extravascular volumes were elevated with no change in cellular volume. The volume of the interstitium available to albumin was more than twice control. The fraction of the interstitium that excludes albumin was calculated to be 0.32 +/- 0.04, with a 51% reduction in absolute excluded volume 2 h after AE. Clearance of hyaluronan by the lymphatics (normalized to baseline) increased 6- to 10-fold during and after AE. It was estimated that < 2% of the total hyaluronan in the lung would be cleared in 24 h under baseline conditions. This amount increased to approximately 11% under AE conditions and approximately 15% under recovery conditions. Changes in lung fluid volumes and protein clearance indicate increased microvascular permeability 2 h after AE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226506 TI - Effects of dehydration on thermoregulatory responses of horses during low intensity exercise. AB - Effects of dehydration on thermoregulatory and metabolic responses were studied in six horses during 40 min of exercise eliciting approximately 40% of maximal O2 consumption and for 30 min after exercise. Horses were exercised while euhydrated (C), 4 h after administration of furosemide (FDH; 1.0 mg/kg i.v.) to induce isotonic dehydration, and after 30 h without water (DDH) to induce hypertonic dehydration. Cardiac output was significantly lower in FDH (144.1 +/- 8.0 l/min) and in DDH (156.6 +/- 6.9 l/min) than in C (173.1 +/- 6.2 l/min) after 30 min of exercise. When DDH, FDH, and C values were compared, dehydration resulted in higher temperatures in the middle gluteal muscle (41.9 +/- 0.3, 41.1 +/- 0.2, and 40.6 +/- 0.2 degrees C, respectively) and pulmonary artery (40.8 +/- 0.3, 40.1 +/ 0.2, and 39.7 +/- 0.2 degrees C, respectively). Temperatures in the superficial thoracic vein and subcutaneous sites on the neck and back and peak sweating rates on the neck and back were not significantly different in DDH and C. In view of higher core temperatures during exercise after dehydration and decrease in cardiac output without concomitant increases in peripheral temperatures or reduced sweating rates, we conclude that the impairment of thermoregulation was primarily due to decreased transfer of heat from core to periphery. PMID- 8226507 TI - Invited editorial on "Peripheral chemoreceptors respond to hypoxia in pontine lesioned fetal lambs in utero". PMID- 8226508 TI - Peripheral chemoreceptors respond to hypoxia in pontine-lesioned fetal lambs in utero. AB - Acute hypoxia inhibits, rather than stimulates, fetal breathing movements (FBM), but there has been controversy as to the activity and role of the peripheral arterial chemoreceptors in the regulation of breathing movements in the unanesthetized fetus in utero. However, after midcollicular brain stem transection or lateral pontine lesion, hypoxia causes FBM to become continuous and stimulated in rate and depth. To determine whether this stimulatory response involves peripheral chemoreceptors, we used a two-stage approach to examine the response to hypoxia after peripheral chemodenervation in lateral pontine-lesioned fetal lambs. The lateral pons was lesioned at 119-121 days, and the response to hypoxia was tested in the unanesthetized fetus 4 days afterward. Fourteen fetuses in which hypoxia stimulated FBM underwent either peripheral chemodenervation or sham denervation in a second operation. Hypoxia had no effect when the fetus was tested 4-5 days after peripheral chemodenervation, and the basal incidence of FBM was significantly lower. The stimulatory response was unchanged by sham denervation. We conclude that the peripheral chemoreceptors are active in fetal life and that they mediate the stimulation of FBM seen in response to hypoxia after removal of the lateral pontine inhibition. In addition, after pontine lesion there is evidence of tonic chemoreceptor-mediated influences on FBM, which are normally overriden in the intact fetus. PMID- 8226509 TI - Contribution of in vivo microvascular PO2 in the cat carotid body chemotransduction. AB - To understand the interplay between microcirculatory control and carotid body (CB) function, we simultaneously measured carotid body microvascular PO2 (CBM PO2) and chemosensory activity in the cat in vivo under several experimental conditions. Cats were anesthetized with pentobarbital sodium, paralyzed, and artificially ventilated. CBs were exposed, and steady-state CBM PO2 was measured by the O2-dependent quenching of the phosphorescence of Pd-meso-tetra-(4 carboxyphenyl)porphine, which was administered intravenously. A few fibers of the carotid sinus nerve were used to record chemosensory discharges. At arterial PO2 (PaO2) of 103.4 +/- 4.1 Torr, CBM PO2 was 52.5 +/- 3.6 Torr (n = 9). Graded lowering of PaO2 from 160 to 50 Torr resulted in nearly proportional decreases in CBM PO2, but at lower PaO2 the decrease in CBM PO2 became more substantial. As PaO2 decreased, chemosensory discharge increased in parallel with CBM PO2. Hypercapnia and hypocapnia did not significantly change the relationship between PaO2 and CBM PO2, although the chemosensory discharge responded significantly. CBM PO2 and chemosensory discharge were not affected by hemorrhagic hypotension until arterial blood pressure fell below approximately 50 Torr and then CBM PO2 decreased and chemosensory discharge increased. The lack of a significant effect of hemorrhagic hypotension indicated that O2 delivery to CB was almost independent of the systemic blood pressure. Taken together, the observations suggest that CB microcirculation and PO2 are subject to control by intrinsic mechanisms and that CBM PO2 is compatible with oxidative metabolism playing a role in O2 chemoreception during hypoxia. PMID- 8226510 TI - Effects of prenatal undernutrition on developing rat diaphragm. AB - The effects of maternal nutritional deprivation (ND) on the proportions and cross sectional areas (CSAs) of type I and II fibers in the developing rat diaphragm (Dia) were examined. In addition, the effects of ND on in vitro contractile and fatigue properties were also studied. Compared with in the control (CTL) group (from pups of well-nourished mothers), proportions of type II fibers were significantly lower in the ND group during the first 3 postnatal wk. However, this difference did not persist into adulthood. CSAs of type I and II fibers were larger in ND Dia than in CTL Dia during early postnatal development, but in adults type I and II fiber CSAs were larger in CTL than in ND Dia. Interstitial space comprised a smaller fraction of total muscle area in ND Dia throughout postnatal development and persisted in adults. Estimated fiber density (no. of fibers/CSA muscle) was lower in ND groups at all ages. In addition, the total number of fibers was significantly lower in ND Dia at all ages. Optimal muscle fiber length for peak twitch and maximum isometric force generation was shorter in ND Dia at all ages. At postnatal days 7 and 14, specific force (maximum tetanic force in N/cm2) of ND Dia was significantly lower than that of CTL Dia. At day 21, Dia specific force was comparable between the two groups but was significantly lower in adult ND compared with adult CTL Dia. Except for days 0 and 14, ND Dia was more fatigue resistant than CTL Dia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226511 TI - Functional electrical stimulation and respiration during sleep. AB - During obstructive sleep apnea (OSA), respiratory activation of upper airway muscles, particularly genioglossus, is ineffective during sleep. Functional electrical stimulation (FES) of muscles reportedly reduces the number and length of OSAs. Our goals were to examine the effect of FES on sensation during wakefulness and on OSA events. Studies were performed in 11 subjects: 4 healthy asymptomatic subjects and 7 patients with OSA. Surface electrodes placed on the submental region produced discomfort; however, during sleep, the stimulus intensity producing arousal was significantly greater than that producing barely tolerable discomfort during wakefulness. Additionally, we developed a protocol for placement of fine-wire electrodes into the neurovascular bundle of the hypoglossal nerve, using recognizable radiographic features and computerized axial tomography as guides. In these patients, while awake, optimal wire placement was associated with visible tongue protrusion without discomfort. With both surface stimulation and fine-wire FES, during sleep the stimulus intensity required to produce obvious electroencephalographic arousal was significantly greater than that producing a barely tolerable sensation while awake. During apneic events, the application of surface stimulation had an inconsistent effect, terminating 22% of the apneas, and fine-wire FES also had a limited impact, terminating 23% of the apneic events. We conclude from our studies that subjects tolerate surface and fine-wire FES to higher stimulus parameters during sleep than during wakefulness but that both approaches have an inconsistent effect on apneas during sleep. PMID- 8226512 TI - Alternative model of respiratory tissue viscoplasticity. AB - Respiratory tissue impedance exhibits both tidal volume and frequency dependences in the ranges of normal breathing. Hildebrandt argued that these indicate tissue viscoplasticity and offered a model in support of his argument consisting of viscoelastic and plastoelastic compartments, both mechanically in parallel (J. Appl. Physiol. 28: 365-372, 1970). Although the model appears to be qualitatively consistent with oscillatory behavior of a wide variety of respiratory tissues, it yields only moderately good quantitative correspondences despite a relatively large number of parameters, eight. One reason may be the model topology, which implies that rate-dependent and amplitude-dependent processes are decoupled. This is contrary to observed behavior. In this study we offer a model in which viscoelastic and plastoelastic compartments are mechanically coupled through a serial arrangement. The total number of parameters in the model is four. Using a least squares technique, we fitted this model to impedance data of chest wall, healthy lungs, and edematous lungs, all measured in vivo. We found that the model could account for the major, as well as the more subtle, features of the chest wall data with fewer parameters and fewer ad hoc assumptions than Hildebrandt's model. Although it lacks anatomic specifics, the model suggests that the observed chest wall behavior may stem from the actin-myosin cross-bridge kinetics. It also seems applicable to lung tissue, although the requirements for the plastoelastic compartment are less certain. In the case of edematous lungs, the applicability of the model is difficult to establish. PMID- 8226513 TI - Maximal lactic capacity at altitude: effect of bicarbonate loading. AB - The aim of the present study was to test the hypothesis that the net maximal blood lactate accumulation ([La]max) during heavy exercise in lowlanders acclimatized to chronic hypoxia may be limited by the reduced bicarbonate stores. Six men [age 32 +/- 4 (SD) yr] performed supramaximal exercise until voluntary exhaustion at sea level (204 +/- 54 W) and after sojourning for 1 mo at 5,050 m (175 +/- 23 W), without (C) and with (B) oral sodium-bicarbonate loading (0.3 g/kg body wt). Exhaustion time, arterial blood lactate concentration, arterial pH (pHa), arterial PCO2, and intramuscular pH were measured at rest and after exercise. At sea level, exhaustion time increased from 6.5 +/- 2.8 min in C to 7.5 +/- 2.7 min in B (P < 0.05). At altitude, exhaustion times were similar to the sea level C values and the same in C and B. At sea level, resting pHa increased from 7.41 +/- 0.02 in C to 7.46 +/- 0.03 in B (P < 0.001); the corresponding values at altitude were 7.46 +/- 0.04 and 7.55 +/- 0.03 (P < 0.001). Postexercise pHa at sea level was 7.22 +/- 0.02 in C and 7.25 +/- 0.08 in B (NS). After exercise at altitude, pHa was 7.32 +/- 0.04 and 7.44 +/- 0.03 in C and B, respectively (P < 0.001). [La]max increased from 12.86 +/- 1.45 mM in C to 16.63 +/- 1.76 mM in B (P < 0.01) at sea level and from 6.85 +/- 1.40 mM in C to 7.95 +/- 1.74 mM in B (NS) at altitude.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226514 TI - NIP-121 is more effective than nifedipine in acutely reversing chronic pulmonary hypertension. AB - To determine if NIP-121, a new antihypertensive agent with K+ channel-opening activity, would be an effective vasodilator in pulmonary hypertension, we studied its acute hemodynamic effects under normoxic conditions in conscious chronically hypoxic pulmonary hypertensive rats and in control pulmonary normotensive rats. In contrast to no pulmonary vasodilation by NIP-121 in control rats, the K+ channel activator (10-100 mg/kg i.v.) decreased both mean pulmonary arterial pressure (from 42 +/- 2 to 33 +/- 2 mmHg; P < 0.05) and total pulmonary resistance (from 278 +/- 30 to 213 +/- 32 mmHg.l-1 x min; P < 0.05) in hypertensive rats. NIP-121 produced similar dose-related decreases in mean systemic arterial pressure and total systemic resistance in both groups of rats. Both the pulmonary and the systemic vasodilations to NIP-121 were inhibited by pretreatment with the blocker of ATP-sensitive K+ channels, glibenclamide (20 mg/kg), but not with the inhibitor of endothelium-derived relaxing factor synthesis, nitro-L-arginine (10 mg/kg). The L-type voltage-gated Ca2+ channel blocker, nifedipine (10-1,000 mg/kg i.v.), failed to cause pulmonary vasodilation in normoxic hypertensive rats, although there was dose-related systemic vasodilation. These results show that in contrast to the Ca2+ channel blocker, nifedipine, the K+ channel activator, NIP-121, is a potent vasodilator of chronic hypoxia-induced pulmonary hypertension in the rat. The mechanism of its hypotensive action in the hypertensive pulmonary vasculature might be more than simply membrane hyperpolarization and indirect inhibition of the L-type voltage gated Ca2+ channel. PMID- 8226515 TI - Reactive oxygen in skeletal muscle. III. Contractility of unfatigued muscle. AB - This study tested the hypothesis that reactive oxygen intermediates present in unfatigued skeletal muscle act to enhance contractile function. Fiber bundles from rat diaphragm were incubated with exogenous catalase (an antioxidant enzyme that dehydrates hydrogen peroxide to molecular oxygen and water) to decrease the tissue concentration of reactive oxygen intermediates. Catalase (10(3) U/ml) significantly decreased twitch characteristics (time to peak tension, half relaxation time, peak force, and twitch-to-tetanus force ratio), thereby shifting the force-frequency relationship to the right. Catalase effects were dose dependent. Concentrations of 1 to 10(5) U/ml progressively depressed submaximal (30-Hz) tetanic stress, whereas concentrations > 10(5) U/ml were toxic, inhibiting maximal (200-Hz) tetanic stress (P < 0.0001). Exogenous hydrogen peroxide (10(-4) to 10(-2)M) increased peak twitch stress (P < 0.03) and lengthened both time to peak tension (P < 0.02) and half-relaxation time (P < 0.02). Selective removal of superoxide anion radicals with the use of superoxide dismutase produced dose-dependent contractile inhibition similar to that produced by catalase. We conclude that the reactive oxygen intermediates present in unfatigued skeletal muscle have a positive effect on excitation-contraction coupling and are obligatory for optimal contractile function. PMID- 8226516 TI - Laryngeal response to passively induced hypocapnia during NREM sleep in normal adult humans. AB - Passively induced hypocapnia in animals activates vocal cord adductor muscles and decreases the glottic aperture. The purpose of this study was to determine if passively induced hypocapnia has similar effects in normal adult humans in stage 3/4 non-rapid-eye-movement (NREM) sleep. Hypocapnia was induced by hyperventilating the subjects with a positive-pressure ventilator via a nose mask. At hypocapnic levels below the CO2 apneic threshold, abrupt cessation of mechanical ventilation was followed by an apnea. In protocol 1, intramuscular electromyographic recordings of intrinsic laryngeal muscles were obtained in nine subjects. Activity of the posterior cricoarytenoid muscle, a vocal cord abductor, disappeared during passive hyperventilation. The muscle remained electrically silent during an apnea, but phasic inspiratory activity reappeared with the first respiratory effort. The thyroarytenoid and arytenoideus muscles, both vocal cord adductors, were electrically silent during spontaneous breathing in NREM sleep. Hypocapnia was frequently associated with activation of both adductor muscles. Once activated, the adductor muscles remained tonically active during an ensuring apnea. In protocol 2, a fiber-optic scope was advanced transnasally into the hypopharynx to determine glottic aperture size during passively induced hypocapnic apnea. In the seven subjects who achieved stable NREM sleep, the glottic aperture during an apnea was smaller than at any time throughout the respiratory cycle during spontaneous breathing just before positive-pressure ventilation. The results suggest that the decrease in glottic aperture observed during an induced hypocapnic apnea is due to suppression of the posterior cricoarytenoid muscle and/or activation of vocal cord adductor muscles. PMID- 8226517 TI - Stress failure of pulmonary capillaries in racehorses with exercise-induced pulmonary hemorrhage. AB - Bleeding into the lungs in thoroughbreds is extremely common; there is evidence that it occurs in essentially all horses in training. However, the mechanism is unknown. We tested the hypothesis that exercise-induced pulmonary hemorrhage (EIPH) is caused by stress failure of pulmonary capillaries. Three thoroughbreds with known EIPH were galloped on a treadmill, and after the horses were killed with intravenous barbiturate the lungs were removed, inflated, and fixed for electron microscopy. Ultrastructural studies showed evidence of stress failure of pulmonary capillaries, including disruptions of the capillary endothelial and alveolar epithelial layers, extensive collections of red blood cells in the alveolar wall interstitium, proteinaceous fluid and red blood cells in the alveolar spaces, interstitial edema, and fluid-filled protrusions of the endothelium into the capillary lumen. The appearances were consistent with the ultrastructural changes we have previously described in rabbit lungs at high capillary transmural pressures. Actual breaks in the endothelium and epithelium were rather difficult to find, and they were frequently associated with platelets and leukocytes that appeared to be plugging the breaks. The paucity of breaks was ascribed to their reversibility when the pressure was lowered and to the fact that 60-70 min elapsed between the gallop and the beginning of lung fixation. Capillary wall stress was calculated from pulmonary vascular pressures measured in a companion study (Jones et al. FASEB J. 6: A2020, 1992) and from measurements of the thickness of the blood-gas barrier and the radius of curvature of the capillaries. The value was as high as 8 x 10(5) dyn/cm2 (8 x 10(4) N/m2), which exceeds the breaking stress of most soft tissues. We conclude that stress failure of pulmonary capillaries is the mechanism of EIPH. PMID- 8226518 TI - Importance of prostanoids in calcium chelator-induced airway constriction. AB - To elucidate the mechanism of calcium chelator-induced airway constriction, we examined the relationship between prostanoids and histamine in bronchoalveolar lavage (BAL) fluid and the magnitude of airway constriction in peripheral airways of anesthetized Basenji-Greyhound dogs. A wedged bronchoscope technique was used to measure collateral system resistance (Rcs) before and after aerosol challenges. Sublobar segments were challenged either with acetylcholine or with Na2EDTA in the presence or absence of meclofenamate sodium (3 mg/kg iv) or methylprednisolone (2 mg.kg-1.day-1). After measurements of Rcs, BAL was performed, and the fluid was analyzed for prostanoids with the use of gas chromatography-mass spectrometry. Sublobar segments challenged with Na2EDTA showed increased concentrations of prostaglandin (PG) D2 but no increases in PGE2, PGF2 alpha, 9 alpha,11 beta-PGF2, 6-keto-PGF1 alpha, thromboxane B2, or histamine. There was a strong relationship (r = 0.84, P = 0.005) between changes in Rcs after Na2EDTA and concentrations of PGD2 in BAL fluid. Acetylcholine, which increased Rcs to a similar degree as Na2EDTA did, produced no significant increase in prostanoid concentrations. Changes in Rcs after Na2EDTA and concentrations of PGD2 were reduced in the presence of meclofenamate or methylprednisolone. These data support the idea that the mechanism of calcium chelator-induced bronchoconstriction involves the release of bronchoconstricting prostanoids. PMID- 8226519 TI - Sea-level PCO2 relates to ventilatory acclimatization at 4,300 m. AB - There is considerable variation among individuals in the extent of, and the time required for, ventilatory acclimatization to altitude. Factors related to this variation are unclear. The present study tested whether interindividual variation in preascent ventilation or magnitude of hypoxic ventilatory response related to ventilatory acclimatization to altitude. Measurements in 37 healthy resting male subjects at sea level indicated a wide range (34-48 Torr) of end-tidal PCO2 values. When these subjects were taken to Pikes Peak, CO (4,300 m, barometric pressure 462 mmHg), the end-tidal PCO2 values measured on arrival and repeatedly over 19 days were correlated with the sea-level end-tidal PCO2. At 4,300 m, subjects with high end-tidal PCO2 had low values of arterial oxygen saturation (SaO2). Also, sea-level end-tidal PCO2 related to SaO2 after 19 days at 4,300 m. Twenty-six of the subjects had measurements of isocapnic hypoxic ventilatory response (HVR) at sea level. The end-tidal PCO2 values on arrival and after 19 days residence at 4,300 m were inversely related to the sea-level HVR values. Thus both the PCO2 and the HVR as measured at sea level related to the extent of subsequent ventilatory acclimatization (decrease in end-tidal PCO2) and the level of oxygenation at altitude. The finding in our cohort of subjects that sea-level end-tidal PCO2 was inversely related to HVR raised the possibility that among individuals the magnitude of the hypoxic drive to breathe influenced the amount of ventilation at all altitudes, including sea level. PMID- 8226520 TI - Hypoxia-induced differential modulation of adenosinergic and muscarinic receptors in rat heart. AB - To better understand the decreased chronotropic response to catecholamines in chronic hypoxia, we compared the inhibitory pathways regulating adenylate cyclase in rats exposed for 30 days to hypobaric hypoxia (380 Torr; HX) with those in control rats (CT) by the analysis of adenosinergic A1-receptors (8-cyclopentyl 1,3-[3H]dipropylxanthine) and muscarinic M2-receptors ([3H]quinuclidinyl benzilate). A1-receptor density was decreased by 46% in sarcolemmal preparations without a change in the affinity for agonist [(R)-phenylisopropyladenosine]. M2 receptor density was increased (HX: 280 +/- 16 fmol/mg, CT: 188 +/- 15 fmol/mg; n = 7; P < 0.001) without a change in dissociation constant. Displacement of [3H]quinuclidinyl benzilate by carbachol indicated significant decreases in the dissociation constants of both superhigh- (HX: 73 +/- 19 nM, CT: 182 +/- 42 nM; P < 0.001) and high-affinity binding sites (HX: 4 +/- 1 microM, CT: 12 +/- 3 microM; P < 0.001). Our data show that chronic hypoxia leads to differential modulation of cardiac receptors with a downregulation of adenosine receptors and increases in muscarinic receptor affinity and density, which may contribute to the blunted responsiveness of the heart to catecholamines. PMID- 8226521 TI - Effects of rapid-eye-movement sleep on the apneic threshold in dogs. AB - We determined whether the apneic threshold after active hyperventilation was different in rapid-eye-movement (REM) vs. non-REM (NREM) sleep. Sleeping dogs were repeatedly exposed to 35-45 s of hypoxia of varying severity (end-tidal PO2 40-60 Torr) that was abruptly terminated with 100% O2. Changes in breathing pattern after brief hypoxia were compared with those after a normoxia-to hyperoxia transition, i.e., control conditions. In NREM sleep, hypoxic hyperventilation was consistently followed by central apnea, the duration of which was linearly related to the corresponding hypocapnia and/or increase in tidal volume (VT) during hypoxia. After hypoxia, expiratory duration averaged 3.5 x control value at -5-Torr change in end-tidal PCO2 and twofold increase in VT; mean expiratory duration was 5 x control value at -10-Torr change in end-tidal PCO2 and fourfold increase in VT. In REM sleep, central apnea of varying duration did occur on occasion after brief hypoxic hyperventilation, but there was no systematic relationship with magnitude of hypocapnia or increase in VT. Breathing pattern during or after hypoxia in REM was not related to temporal changes in either eye movement density or electroencephalogram frequency. Thus, in contrast to NREM sleep, in REM sleep ("phasic" or "tonic") a posthyperventilation apneic threshold was not present. We attribute this effect of REM to 1) a reduced VT response to hypoxia that would minimize inhibitory "memory" effect from lung stretch and 2) attenuated inhibitory response to any given magnitude of hypocapnia or increased VT. Active hyperventilation-induced apneic threshold may be "masked" by actions of nonchemoreceptor and nonmechanoreceptor inputs affecting respiratory motor output in REM sleep. These data are consistent with the relative absence of central apnea and periodic breathing in humans in REM sleep. PMID- 8226522 TI - Effects of short-term and long-term androgen treatment on the diaphragm in male and female rats. AB - The effects of short-term (2.5 wk) and long-term (10 wk) testosterone propionate (2.5 mg/day; 5 days/wk) treatment on diaphragm contractility, fatigue resistance, and fiber type proportions were studied in male and female rats. Contractility and fatigue resistance indexes were measured in an in vitro diaphragm costal strip preparation by direct stimulation at 37 degrees C. The fatigue paradigm consisted of 30 trains/min at 5 Hz (50% duty cycle) for 10 min. Fatigue resistance indexes were calculated as postfatigue divided by baseline forces. In females but not males, testosterone treatment produced significant increases in body weight, costal diaphragm weight, and contractility and significant decreases in fatigue resistance indexes. The interaction between testosterone treatment and the duration of treatment was significant, with the increase in contractility (females) being significant after short-term but not long-term treatment. No significant difference in fiber type proportions or areas was observed, regardless of treatment duration or the preexperimental, basal circulating level of androgen. PMID- 8226523 TI - Structure of parasternal intercostal muscles in the adult hamster: topographic effects. AB - The parasternal intercostals are primary inspiratory muscles like the costal and crural diaphragm. However, the structure of the rib cage and its impedance to inspiration and expiration varies regionally. We questioned whether topographic differences in rib cage structure and impedance were associated with regional differences in parasternal intercostal muscle structure. Therefore, we examined the size and percentage of histochemically stained fibers in the parasternal intercostal muscles in the first, second, third, fourth, and sixth interspaces in the hamster. We observed a rostrocaudal gradient in the percentage and size of slow oxidative (SO), fast oxidative-glycolytic, and fast glycolytic (FG) fibers in the parasternal intercostal muscles. In particular, the percentage of SO decreased while the percentage of FG increased in a rostrocaudal direction in the first through sixth interspaces. In addition, the size of SO and FG fibers increased from the first to sixth interspace. Furthermore, changes in the size and percent of the three fiber types produced, in a rostrocaudal direction, significant reductions in the relative mass of the parasternal intercostal muscle made up of SO fibers and increases in the mass of fast fibers. We speculate that topographical differences in the size and percentage of fast and slow twitch fibers in the parasternal intercostal are likely to alter force-generating capacity of the parasternal muscles in a rostrocaudal direction and likely reflect regional differences in muscle load and/or activity. PMID- 8226524 TI - Dependence of maximal flow-volume curves on time course of preceding inspiration. AB - Thirteen normal subjects, sitting in a body plethysmograph and breathing through a pneumotachograph, performed forced vital capacity maneuvers after a rapid inspiration without or with an end-inspiratory pause (maneuvers 1 and 2) and after a slow inspiration without or with an end-inspiratory pause (maneuvers 3 and 4), the pause lasting 4-6 s. Inspirations were initiated close to functional residual capacity. At all lung volumes, expiratory flow was larger with maneuver 1 than with any other maneuver and, over the upper volume range, larger with maneuver 3 than with maneuver 4, whereas it was similar for maneuvers 2 and 4. Relative to corresponding values with maneuver 4, peak expiratory flow was approximately 16 and approximately 4% larger with maneuvers 1 and 3, respectively, whereas forced expiratory volume in 1 s increased by approximately 5% only with maneuver 1. The time dependence of maximal flow-volume curves is consistent with the presence of viscoelastic elements within the respiratory system (D'Angelo et al. J. Appl. Physiol. 70: 2602-2610, 1991). PMID- 8226525 TI - The development of aerobic power in young athletes. AB - Previous studies investigating the effects of training in children have been hampered in their interpretation by the confounding effects of growth and development. We followed the development of maximal aerobic power (VO2max) in 453 athletes drawn from soccer, swimming, gymnastics, and tennis. Study design was of a mixed longitudinal type with five age cohorts (8, 10, 12, 14 and 16 yr) followed for 3 consecutive years. A multilevel regression modeling procedure was used to identify the independent effects of predictor variables while accounting for the effects of growth, such as changes in body size. When age, height, and weight were controlled for, VO2max in males significantly increased with pubertal status, indicated by the coefficient value of 0.15 l/min being greater than its associated SE of 0.07 l/min. Females showed a similar pattern, with a coefficient value of 0.13 +/- 0.07 l/min, although the significant increase in VO2max (P < 0.05) found in males in the latter stages of puberty was not shown in females. Swimmers had the highest VO2max values (P < 0.001) at all ages. PMID- 8226526 TI - Washout of protein in dog lung lymph. AB - After an increase in microvascular filtration rate, lung lymph may contain protein washed from the tissue spaces plus protein from the filtrate. If so, then the lymphatic protein concentration may be significantly higher than the filtrate protein concentration (Cf). To test this hypothesis, we decreased the plasma protein concentration from 5.1 +/- 0.6 to 0.54 +/- 0.15 g/dl and increased the pulmonary microvascular filtration rate in four dogs. We estimated Cf to be 0.16 +/- 0.05 g/dl after we reduced the plasma protein concentration, and the lymphatic protein concentration (0.43 +/- 0.04 g/dl) was significantly greater than Cf. Our results indicate that lung microvascular membrane reflection coefficients estimated from lung lymph data may be too low. However, the amount of error caused by tissue protein washout is probably small. To account for the protein washout error, we estimated the lung microvascular membrane reflection coefficient to be approximately 0.74-0.76 instead of the approximately 0.70 previously reported for dogs (J. C. Gabel et al. J. Appl. Physiol. 55: 866-869, 1983). PMID- 8226527 TI - Urea kinetics in humans at two levels of exercise intensity. AB - A primed constant infusion of [15N2]urea was used to quantify the response of urea production to exercise at 40 and 70% maximal oxygen consumption on a treadmill. Total urea production, urea production from recycled N, urea production from nonrecycled N, and urea N recycled back into body protein were calculated. Most components of urea kinetics were unaffected by exercise at either intensity. The rate of urea reincorporated into protein was significantly increased during exercise and recovery at both levels of exercise. We conclude that exercise does not stimulate urea production but that there may be an accelerated reincorporation of urea N back into body protein. PMID- 8226528 TI - Carbonic anhydrases in cytosol, nucleus, and membranes of rat liver. AB - The relative contribution of each functional carbonic anhydrase (CA) isozyme to liver CA activity of fed or starved adult male rats has been determined. The functional isozymes are CA II, CA III, CA IV, and CA V. Total CA, CA III, CA II, CA IV, and CA V activities (in mumol CO2 converted.min-1.liver-1), as measured by mass spectrometric assay using NaHC18O16O in aqueous solution at pH 7.4 and 37 degrees C, were 94,867, 38,621, 37,000, 14,515, and < 5,000 in fed rats and 40,630, 10,498, 9,137, 18,338, and < 2,600 in starved rats, respectively. CA II was unevenly distributed throughout the liver. In perivenous and periportal cytosols, as determined by the digitonin-pulse perfusion technique, CA II activity was (in mg cytosolic protein-1) 325 and 69 in fed rats and 167 and 33 in starved rats, respectively. CA III was more evenly distributed and less affected by starvation: CA III activity in perivenous and periportal cytosols was (in mg cytosolic protein-1) 84 and 55 in fed rats and 113 and 52 in starved rats, respectively. Evidence that CA III was concentrated in the nucleus was obtained histochemically by the Ridderstrale cobalt-precipitation technique in 2-microns thick glutaraldehyde-fixed sections from adult fed rats. Liver CA activity was higher in the perivenous hepatocytes in cytosols and nuclei, whereas CA IV was homogeneously distributed. Incubation of the 2-microns sections with 1 microM acetazolamide resulted in inhibition of all membrane-associated CA, 50% of cytosolic CA, and no nuclear CA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226529 TI - Effect of prolonged alveolar hypoxia on pulmonary arterial pressure and segmental vascular resistance. AB - The time course of hypoxic pulmonary vasoconstriction and its segmental distribution were studied during prolonged (150 min) alveolar hypoxia in in vivo dog lungs at constant-flow perfusion. With the pulmonary and the systemic circulations separated by two extracorporeal circuits, adequate systemic oxygenation was achieved throughout the experiments. The pulmonary circulation exhibited a time-related biphasic hypoxic vasoconstrictor response: an initial rapid contraction [79 +/- 11% (SE) above control level] was followed by a partial relaxation when a second slow and sustained vasoconstriction (92 +/- 13% above control level) superseded. We partitioned the pulmonary circulation into two segments by arterial occlusion: an upstream arterial segment and a downstream segment consisting of a middle and a venous segment. Measurements were performed at baseline and during the late sustained vasoconstrictor response. Prolonged alveolar hypoxia increased pulmonary capillary pressure by 90 +/- 18%, the site of pulmonary vasomotion being the arterial upstream and downstream middle and venous segments. PMID- 8226530 TI - Heart function after injection of small air bubbles in coronary artery of pigs. AB - By its nature, vaporization of atherosclerotic plaques by laser irradiation or spark erosion may produce a substantial amount of gas. To evaluate the effect of gas embolism possibly caused by vaporization techniques, air bubbles with diameters of 75, 150, or 300 microns, each in a volume of 2 microliters/kg, were selectively injected subproximal in the left anterior descending coronary artery of seven anesthetized pigs (28 +/- 3 kg). Systemic hemodynamics such as heart rate, left ventricular pressure and its peak positive first derivative, and mean arterial pressure did not change after air injection, whereas there was a minor change in peak negative first derivative of left ventricular pressure. After injection of air bubbles there was a maximal relative reduction of systolic segment shortening (SS) in the myocardium supplied by the left anterior descending coronary artery of 27, 45, and 58% for 75-, 150-, and 300-microns bubbles, respectively, and a relative increase of postsystolic SS (PSS) of 148, 200, and 257% for 75-, 150-, and 300-microns bubbles, respectively. Recovery of SS and PSS started after 2 min and was completed after 10 min. A difference in SS and PSS changes between different bubble size injections could be demonstrated. From this study it is clear that depression of regional myocardial function after injection of air bubbles could pass unnoticed on the basis of global hemodynamic measurements. PMID- 8226531 TI - Effects of altitude acclimatization on fluid regulatory hormone response to submaximal exercise. AB - To determine the effect of altitude acclimatization on plasma levels of atrial natriuretic peptide (ANP) during submaximal exercise and its relationship with renin and aldosterone, seven male volunteers aged 17-23 yr exercised to exhaustion on a cycle ergometer at 80-85% of their maximum O2 uptake at sea level (SL; 50 m), during 1 h in a hypobaric chamber [acute altitude (AA); 4,300 m], and after 14 or 16 days of residence on the summit of Pikes Peak, CO [chronic altitude (CA); 4,300 m]. Plasma samples taken before exercise, 10 min after the start of exercise, and 5 min postexercise were analyzed for ANP, plasma renin activity (PRA), and aldosterone (ALDO). ANP showed a progressive increase from rest to postexercise [7.49 +/- 1.63 to 11.32 +/- 1.80 (SE) pmol/ml and 6.05 +/- 2.55 to 10.38 +/- 7.20 pmol/ml; P = 0.049, exercise] at SL and AA, respectively, but not at CA (P = 0.039, altitude). Similarly, PRA and ALDO rose from rest to postexercise (P < 0.001, exercise), but the rise in ALDO with exercise was less during AA than during SL and CA (P = 0.002, phase). The decreased ANP levels during exercise after altitude acclimatization, with no change in PRA and ALDO, suggest that ANP has little effect on PRA and ALDO under these conditions. PMID- 8226532 TI - Neurogenic modulation of periodic hemodynamics in rabbit skeletal muscle. AB - We characterized the effect of various forms of neural blockade and vasopressin infusion on regular slow-wave flux oscillation (RSWFO). RSWFO, measured with laser-Doppler flowmetry (LDF), was induced by local reduction of arterial pressure in the gastrocnemius muscle of 30 New Zealand White rabbits. At normal median femoral arterial mean pressure, LDF only showed irregular variations. With an intact innervation, RSWFO was maximal at a median femoral mean pressure of 30 mmHg (range 25-53), the median frequency was 1.7 cycles/min (range 1.0-3.0), and the maximum amplitude was 50% (range 19-119). Application of lidocaine to the sciatic nerve, or cutting of this nerve, when arterial pressure was reduced, resulted in disappearance of slow waves in all cases. Intravenous application of guanethidine resulted in a significant reduction of the flux oscillation amplitude from 47 to 10% (P < 0.001). In an additional nine animals with sciatic denervation and local blood pressure reduction, RSWFO could be induced by intravenous vasopressin infusion. These results offer a new pathophysiological concept of periodic hemodynamics as a locally controlled and, with intact innervation, a neurally modulated mechanism for blood flow redistribution in cases of reduced blood pressure and flow. PMID- 8226533 TI - Effects of continuous negative airway pressure-related lung deflation on upper airway collapsibility. AB - Continuous negative airway pressure (CNAP) causes a decrease in lung volume, which is known to increase upper airway resistance by itself. We studied how this lung volume change could modify upper airway collapsibility with five normal awake subjects. In a first trial, pressure in a nasal mask (Pm) was progressively decreased in 3- to 5-cmH2O steps (CNAP). In a second trial, changes in lung volumes resulting from CNAP were prevented by applying simultaneously an equivalent level of negative extrathoracic pressure into a poncho-type respirator [isovolumetric CNAP (CNAPisovol)]. For each trial, we examined the relationship between the maximal inspiratory airflow of each flow-limited inspiratory cycle and the corresponding Pm by least-squares linear regression analysis and determined the critical pressure. We also determined the Pm threshold corresponding to the first Pm value below which flow limitation occurred. Flow limitation was observed in each subject with CNAP but in only two subjects with CNAPisovol. In these two subjects, the Pm threshold values were -20 and -9 cmH2O with CNAP and -39 and -16 cmH2O with CNAPisovol, respectively. Critical pressures for the same two subjects were -161 and -96 cmH2O with CNAP and -202 and -197 cmH2O with CNAPisovol, respectively. We conclude that CNAP-induced decreases in lung volume increase upper airway collapsibility. PMID- 8226534 TI - Role of protein intake on protein synthesis and fiber distribution in the unweighted soleus muscle. AB - Protein turnover in skeletal muscle is very sensitive to protein intake. To examine whether protein intake is able to affect protein synthesis in the atrophied soleus muscle, the effects of a high-protein (30%, HP) and a medium protein (15%, MP) diet were studied in rats after 21 days of hindlimb unweighting. Three weeks of unweighting induced a sharp decrease in food intake (30%). The fractional rate of protein synthesis (ks) was determined in vivo in the slow-twitch soleus muscle by use of a flooding-dose method. With respect to pair-fed animals, a significant reduction in ks occurred (33%) in MP non-weight bearing rats, whereas it was of lesser magnitude and not significant in HP rats. In the atrophied soleus muscle of non-weight-bearing MP rats, a large decrease (42%) in type I fiber distribution was accompanied by an increase in intermediate and type IIa fibers. By contrast, a higher percentage of type I fiber was maintained with the HP diet. However, the HP diet had no beneficial effect in preventing the decrease in either type I fiber cross-sectional area (65%) or the average decrease in absolute myofibrillar and mitochondrial volumes (69 and 52%, respectively). These results demonstrate that an HP intake did not prevent soleus muscle atrophy but may sustain protein synthesis and partly preserve fiber type distribution without affecting the ultrastructural composition of fibers. Because the circulating level of free 3,5,3'-triiodothyronine was reduced by 14% with the HP diet, this effect on fiber type distribution, and possibly protein synthesis, may involve thyroid hormones. PMID- 8226535 TI - Effects of oxygen tension and glucose concentration on ischemic injury in ventilated ferret lungs. AB - In the ventilated ischemic lung, oxygen tension will increase at a time when glucose depletion may impair antioxidant defenses, thereby predisposing the lung to injury mediated by oxygen radicals. In the unventilated ischemic lung, however, glucose depletion in the setting of low oxygen tension may decrease production of ATP, leading to injury by a different mechanism. In this study, we evaluated the role of both oxygen tension and glucose concentration on ischemic injury in isolated ferret lungs. Injury, defined as an increase in vascular permeability, was assessed by measurement of filtration coefficient (Kf) and osmotic reflection coefficient for albumin (sigma alb) after 3 h of normothermic (37 degrees C) ischemia without reperfusion. Lungs were ventilated with either 95% O2-5% CO2 or 0% O2-5% CO2. The vasculature was flushed with physiological salt solution containing either 15 mM glucose (hyperoxia-glucose, anoxia glucose), 15 mM sucrose (hyperoxia-sucrose, anoxia-sucrose), or no substrate (hyperoxia-no substrate, anoxia-no substrate) (n = 6 for each condition). Kf and sigma alb in hyperoxia-no substrate group did not differ from values in minimally ischemic normoxic normoglycemic ferret lungs. Without glucose, ischemic injury was worse in anoxic than in hyperoxic lungs. With glucose, ischemic injury was worse in hyperoxic than in anoxic lungs. Glucose exacerbated injury in hyperoxic, but not anoxic, lungs. These results indicate that ischemic injury in these lungs depended on both oxygen tension and glucose concentration and suggest that both oxygen radical generation and ATP depletion during ischemia may contribute to the development of this injury. PMID- 8226536 TI - Effect of catheter flow direction on CO2 removal during tracheal gas insufflation in dogs. AB - Tracheal gas insufflation (TGI) improves the efficiency of CO2 elimination by replacing CO2 in the anatomic dead space proximal to the catheter tip with fresh gas during expiration. Turbulence generated by gas exiting the catheter tip may also contribute to alveolar ventilation. To separate distal (turbulence-related) and proximal (washout of dead space) effects of TGI, we compared the efficacy of a straight and an inverted catheter during continuous and expiratory TGI in six mechanically ventilated dogs. We reasoned that the inverted catheter cannot improve CO2 elimination from more distal conducting airways. During continuous TGI with the straight catheter, arterial PCO2 (PaCO2) decreased significantly from baseline (without TGI) of 56 +/- 10 Torr to 38 +/- 8, 36 +/- 8, and 35 +/- 8 Torr at catheter flow rates (Vcath) of 5, 10, and 15 l/min, respectively. For the same conditions, PaCO2 was always higher (P < 0.001) with the inverted catheter (42 +/- 10, 41 +/- 10, and 41 +/- 10 Torr). PaCO2 was lower with the straight (40 +/- 9 Torr) than with the inverted catheter (44 +/- 10 Torr, P < 0.001) during TGI delivered only during expiration at a Vcath of 10 l/min. End-expiratory lung volume relative to baseline increased during continuous, but not during expiratory, TGI and was significantly greater with the straight than with the inverted catheter (P < 0.0001). Our data confirm that the primary mechanism of TGI is expiratory washout of the proximal anatomic dead space but also suggest a minor contribution of turbulence beyond the tip of the straight catheter. PMID- 8226537 TI - Breathing route and resistive loading: influence on ventilatory response to hypoxia in conscious dogs. AB - Unlike normal humans, the tracheostomized conscious dog does not show ventilatory adaptation in response to sustained isocapnic hypoxia. To determine whether this phenomenon is a result of the breathing route or the relatively low airflow resistance of tracheostomy breathing, we evaluated the ventilatory response to sustained isocapnic hypoxia (20 min; arterial oxyhemoglobin saturation = 80%) in five awake dogs during nasal-oral (mask) breathing, tracheal breathing, and tracheal breathing with added matched resistance of upper airway breathing. Mask breathing, like unloaded tracheal breathing, was associated with a consistent level of hyperventilation during the entire hypoxic exposure period. However, mask breathing was always less (P < 0.05) than that found during unloaded tracheal breathing. Loaded tracheal breathing during hypoxia resulted in initial hyperventilation similar to that of unloaded tracheal breathing followed by a "roll off" to a lower minute ventilation similar to that of mask breathing. Our findings demonstrate that ventilatory adaptation is only present during loaded tracheal breathing in dogs and suggest that the breathing route and upper airway resistive loading may play roles in ventilatory adaptation. PMID- 8226538 TI - In vivo antioxidant gene expression in human airway epithelium of normal individuals exposed to 100% O2. AB - Human bronchial epithelium is exquisitely sensitive to high O2 levels, with tracheobronchitis usually developing after 12 h of exposure to 100% O2. To evaluate whether this vulnerability results from inability of the bronchial epithelium to provide adequate antioxidant protection, we quantified antioxidant gene expression in bronchial epithelium of normal volunteers at baseline and after exposure to 100% O2 in vivo. After 14.8 +/- 0.2 h of 100% O2, 24 of 33 individuals had evidence of tracheobronchitis. Baseline gene expression of CuZn superoxide dismutase (SOD), MnSOD, and catalase in bronchial epithelium was very low (CuZnSOD 4.1 +/- 0.8 transcripts/cell, MnSOD 5.1 +/- 0.9, catalase 1.3 +/- 0.2), with control gamma-actin expression relatively abundant (50 +/- 6 transcripts/cell). Importantly, despite 100% O2 exposure sufficient to cause tracheobronchitis in most individuals, antioxidant mRNA transcripts/cell in bronchial epithelium did not increase (P > 0.5). Catalase activity in bronchial epithelium did not change after exposure to hyperoxia (P > 0.05). Total SOD activity increased mildly (P < 0.01) but not sufficiently to protect the epithelium. Together, the very low levels of expression of intracellular antioxidant enzymes and the inability to upregulate expression at the mRNA level with oxidant stress likely have a role in human airway epithelium susceptibility to hyperoxia. PMID- 8226539 TI - Progressive stretch overload of skeletal muscle results in hypertrophy before hyperplasia. AB - Intermittent stretch of the anterior latissimus dorsi (ALD) muscle produces fiber hypertrophy without fiber hyperplasia (J. Appl. Physiol. 74: 1893-1898, 1993). This study was undertaken to determine if a progressive increase in load and duration of stretch would induce extremely large muscle fiber areas or if the fibers would reach a critical size before the onset of fiber hyperplasia. Weights ranging from 10 to 35% of the bird's mass were attached to the right wing of 26 adult quail while the left wing served as the intra-animal control. The stretch protocol was as follows: day 1 (10% wt), days 2 and 3 (rest), day 4 (15% wt), days 5-7 (rest), day 8 (20% wt), days 9 and 10 (rest), days 11-14 (25% wt), days 15 and 16 (rest), and days 17-38 (35% wt). Birds were killed after 12, 16, 20, 24, and 28 days of stretch not including rest days. Muscle mass increased 174% (12 days), 196% (16 days), 225% (20 days), 264% (24 days), and 318% (28 days). Muscle length increased 60% (12 days), 34% (16 days), 59% (20 days), 50% (24 days), and 51% (28 days). Mean fiber area increased 111% (12 days), 142% (16 days), 75% (20 days), 90% (24 days), and 39% (28 days). Fiber number, which was measured histologically, increased significantly by 82% only in the 28 days of stretch group. The percentage of slow tonic fibers did not change for any of the time points examined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226540 TI - Effect of concentric exercise on serum muscle and collagen markers. AB - The effect of an acute bout of high-intensity concentric exercise on serum muscle and collagen marker proteins was studied in nine male students. The muscle derived serum carbonic anhydrase III, myoglobin, and creatine kinase all increased as a result of the exercise. Serum type I procollagen carboxyterminal propeptide decreased at first but started to increase 2 days after the exercise. Serum galactosylhydroxylysyl glucosyltransferase was elevated immediately after the exercise. No significant changes were seen in the concentrations of serum amino-terminal propeptide of type III procollagen or 4-hydroxyproline. It seems that a single bout of heavy concentric exercise causes protein leakage from muscles and probably from the collagen-synthesizing cells of the connective tissue, which may be accompanied by an initial decrease and a subsequent increase in type I collagen production. The activation of type I collagen production seems to depend on the strain and damage of the musculoskeletal system. PMID- 8226541 TI - Inhaled nitric oxide: dose response and the effects of blood in the isolated rat lung. AB - Inhaled nitric oxide (NO) is a vasodilator selective to the pulmonary circulation. Using isolated rat lungs, we determined the dose-response relationship of NO and the role of blood in mediating pulmonary vasodilation and selectivity. Inhaled 20, 50, 100, and 1,000 ppm NO attenuated (P < 0.001) hypoxic pulmonary vasoconstriction by 16.1 +/- 4.9, 22.6 +/- 6.8, 28.4 +/- 3.5, and 69.3 +/- 4.2%, respectively. Inhaled 13, 34, 67, and 670 ppm NO attenuated the increase in pulmonary arterial pressure secondary to angiotensin II more (P < 0.001) in Greenberg-Bohr buffer- (GB) than in blood-perfused lungs (51.7 +/- 9.9, 71.9 +/- 8.9, 78.2 +/- 5.3, and 91.9 +/- 2.1% vs. 14.3 +/- 4.1, 23.8 +/- 4.6, 28.4 +/- 3.8, and 55.5 +/- 5.9%, respectively). Samples from GB- but not blood perfused lungs contained NO (93.0 +/- 26.3 nM). Intravascular NO attenuated the response to angiotensin II more (P < 0.001) in GB- (with and without plasma) than in blood- (hematocrit = 41 and 5%) perfused lungs (75.6 +/- 6.4 and 70.9 +/- 4.8% vs. 22.2 +/- 2.4 and 39.4 +/- 7.6%). In conclusion, inhaled NO produces reversible dose-dependent pulmonary vasodilation over a large range of concentrations. Inhaled NO enters the circulation, but red blood cells prevent systemic vasodilation and also a significant amount of pulmonary vasodilation. PMID- 8226542 TI - Tissue gas stores of the body and head-out immersion in humans. AB - Breath-by-breath gas exchange was studied in 10 subjects during and after transitions between dry conditions and head-out immersion in thermoneutral conditions. Cardiac index (CI) was estimated by means of impedance cardiography. Previous largely qualitative models of changes in tissue gas stores after blood volume shifts could be confirmed and extended to include a quantitative analysis of O2 and CO2 tissue stores. An increase in CI by 47.0% during immersion was associated with an increase in the tissue O2 stores by 122 ml/m2 and a decrease in the tissue CO2 stores by 148 ml/m2. The time constants for the recovery of O2 uptake (tau O2) and CO2 elimination after initial increases after the dry-to immersion transition were 32.4 and 79.3 s, respectively. The decrease in CI on return to the dry conditions was associated with a drop in tissue O2 stores and a tau O2 of 144 s. The increase in tissue O2 stores during immersion as well as the difference in tau O2 between the two transitions were larger than could be explained by the change in CI only. This was attributed to changes in the distributions of peripheral blood flow and venous blood volume. Compared with the O2 stores, the decrease in CO2 stores was better predicted by the change in CI. The present results emphasize that the changes in pulmonary and tissue gas exchange imposed by head-out immersion transients mainly reflect movement of gas in and out of body stores. PMID- 8226543 TI - Tenotomy and repair of latissimus dorsi muscles in rats: implications for transposed muscle grafts. AB - The functional properties of latissimus dorsi (LTD) muscles were evaluated 160 to 180 days after tenotomy and repair, when grafts had stabilized. Our hypothesis was that, compared with control LTD muscles, LTD grafts would develop less absolute force and power but that the specific force and normalized power would not differ. Expressed as a percentage of the value for control LTD muscles, values for grafts were 67% for muscle mass, 74% for mean single fiber cross sectional area, 56% for maximum absolute isometric tetanic force, 64% for maximum absolute average force during shortening, and 70% for maximum absolute power. Compared with control LTD muscles, grafts showed no significant differences either in the number of fibers in the total muscle cross section or in the optimum velocity for the development of power. When force and power of grafts were normalized for total fiber cross-sectional area and mass, respectively, only the value for maximum specific force (84% of control value) was significant. The mechanisms responsible for the decrease in specific force after tenotomy and repair are not known. In contrast to the deficit in maximum specific force, the 30% deficit in maximum absolute power of grafts compared with control LTD muscles was explained completely by the 33% smaller muscle mass. PMID- 8226544 TI - Effects of indomethacin on newborn pig pial arteriolar responses to PCO2. AB - The present experiments were designed to determine whether hypocapnic cerebral vasoconstriction, like hypercapnic dilation, involves prostanoids and, if not, whether alternative mechanisms are related to the absolute arterial PCO2 (PaCO2) or the direction of change. We determined effects of indomethacin (5 mg/kg iv) on pial arteriolar responses to 1) increased PCO2 from normal, 2) decreased PCO2 from normal, and 3) increased PCO2 from hypocapnia to normocapnia in anesthetized newborn pigs. Pial arterioles constricted in response to hypocapnia (PaCO2 = 15 24 Torr) similarly before (-13 +/- 3%) and after (-16 +/- 2%) indomethacin. Cortical periarachnoid cerebrospinal fluid prostanoids were not increased by hypocapnia. As previously reported, cerebral vascular responses to hypercapnia (which increases cerebrospinal fluid prostanoids) were lost after indomethacin. To determine whether the failure of indomethacin to affect the responses to hypocapnia was due to the direction of change (decreasing) or the absolute level of PCO2, piglets were hyperventilated to approximately 15 Torr PaCO2. Increasing PaCO2 in these piglets to approximately 44 Torr caused pial arteriolar dilation (46 +/- 7%) that was not blocked by indomethacin (33 +/- 5%). Cortical periarachnoid prostanoids were not altered when PaCO2 was raised from hypocapnia to normocapnia. Therefore the relationship between CO2 and piglet cerebral vascular tone appears to involve multiple mechanisms. Specifically, dilation in response to CO2 above the normal range appears to involve prostanoids but changes in pial arteriolar diameter at low PaCO2 do not. PMID- 8226545 TI - Effect of inspired CO2 on ventilation and perfusion heterogeneity in hyperventilated dogs. AB - We studied the effect of inspired CO2 on ventilation-perfusion (VA/Q) heterogeneity in dogs hyperventilated under two different tidal volume (VT) and respiratory rate conditions with the use of the multiple inert gas elimination technique. Dogs anesthetized with pentobarbital sodium were hyperventilated with an inspired fraction of O2 of 0.21 by using an increased VT (VT = 30 ml/kg at 18 breaths/min) or an increased respiratory rate (VT = 18 ml/kg at 35 breaths/min). The arterial CO2 tension (PaCO2) was varied to three levels (20, 35, and 52 Torr) by altering the inspired PCO2. The orders of type of ventilation and PaCO2 level were randomized. Compared with normocapnia, VA/Q heterogeneity was increased during hypocapnia induced by increased respiratory rate ventilation, which was indicated by an increase in dispersion indexes and arterial-alveolar inert gas partial pressure difference areas (P < 0.01). In contrast, VA/Q heterogeneity was not affected by hypocapnia when a large VT ventilation was used. Under the conditions of our study, hypercapnia did not result in statistically significant changes in VA/Q heterogeneity with either type of ventilation. Increased VT ventilation reduced dead space at all PaCO2 levels (P < 0.01) and reduced the log standard deviation of the ventilation distribution during normocapnia (P < 0.05) and hypocapnia (P < 0.01). We conclude that hypocapnia increased VA/Q heterogeneity when hyperventilation was achieved with a rapid respiratory rate. Therefore, a lack of improvement in VA/Q matching with inhaled CO2 may be associated with the use of a large VT. These data suggest that hypocapnic bronchoconstriction may be important in mediating hypocapnia-induced VA/Q inequality in dogs. PMID- 8226546 TI - Effect of ozone exposure in vivo on response of bronchial rings in vitro: role of intracellular Ca2+. AB - In this study we investigated the role of intracellular Ca2+ in ozone- (O3) induced airway hyperresponsiveness. Acetylcholine-induced airway responses were measured before and after inhalation of O3 (3 ppm, 30 min) or dry air. In vitro experiments were performed with intact ring segments of third- to fifth-order bronchi. Bronchial responses to carbachol (CCh) were evaluated in Krebs solution (2.5 mM Ca2+) and in Ca(2+)-free [0.1 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA)] Krebs solution with or without indomethacin (IDM, 10(-5) M) and were expressed as percentage of the maximal KCl response (60 mM). Inhalation of O3, but not dry air, caused airway hyperresponsiveness to acetylcholine in vivo. Responses to 50% effective concentrations of CCh were similar in bronchial preparations from O3 and control animals (with or without IDM) in normal Krebs solution. In Ca(2+)-free solution, CCh induced a sustained (20-min) bronchial contraction. These contractions relaxed immediately when nifedipine or a high EGTA concentration was added to the organ bath. The sustained contraction was abolished when the tissues had been incubated with cyclopiazonic acid (10(-5) M), a novel inhibitor of the sarcoplasmic reticulum Ca2+ pump. After O3 exposure, responses of the bronchial smooth muscle (in Ca(2+)-free medium without IDM) were increased (P < 0.05) compared with controls during the first and second CCh stimulations. This O3 induced increase in response to CCh in Ca(2+)-free solution was abolished when the tissues were incubated with IDM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226547 TI - Role of pulmonary surfactant in airway closure: a computational study. AB - A numerical model that simulates airway closure by liquid bridging during expiration has been developed. The effects of both surfactant and time-varying geometry have been included; the model determines the liquid layer flow resulting from a surface tension (Rayleigh) instability, and the computation traces the film's development to closure, yielding pressure, velocity, surface shape, and surfactant concentration distributions. It is found that surfactant is effective in retarding or eliminating liquid bridging through the reduction of the mean surface tension and the action of surface tension gradients. The former effect is also critical in minimizing the magnitude of the negative pressure in the liquid layer and thus presumably in reducing the tendency for airway compliant collapse. PMID- 8226548 TI - Cardiovascular responses to voluntary and treadmill exercise in rats. AB - Male Sprague-Dawley rats (n = 12) were housed in activity wheels and familiarized with treadmill running 2 wk before they were instrumented with Doppler flow probes and a carotid catheter. Mean arterial pressure (MAP), heart rate (HR), mesenteric blood flow (BFmes), and iliac blood flow were determined during bouts of voluntary and treadmill exercise. One voluntary exercise bout (speed = 33 +/- 2 m/min, duration = 26 +/- 5 s) from each rat was selected to compare with 30 s of treadmill exercise at 30 m/min. Voluntary exercise produced increases in MAP (7 +/- 3 mmHg) and HR (63 +/- 7 beats/min) that were significantly less than the increases of 21 +/- 5 mmHg and 95 +/- 9 beats/min, respectively, with treadmill exercise. Voluntary exercise caused an immediate reduction in BFmes of 32 +/- 6%, whereas treadmill exercise produced a significantly greater reduction of 57 +/- 4%. Voluntary and treadmill exercise caused similar increases in iliac blood flow of 112 +/- 15 and 169 +/- 31%, respectively. The patterns of cardiovascular adjustments to the initiation of voluntary exercise are similar to those observed at the initiation of treadmill exercise; however, MAP, HR, and BFmes responses were significantly greater with treadmill exercise. PMID- 8226549 TI - Respiratory muscle fatigue resistance relates to myosin phenotype and SDH activity during development. AB - We correlated the fatigue resistance (FR) of the costal diaphragm (DIA) and external abdominal oblique (EAO) of the rat during postnatal development with their respective 1) myosin heavy chain (MHC) phenotypes and 2) oxidative capacities [indexed by quantitative measurements of succinic dehydrogenase (SDH) enzyme activity]. FR was measured in vitro during isometric contractions with the use of the Burke fatigue test. FR of the DIA and EAO was high in newborns and declined during postnatal development. SDH activity was uniformly low in neonatal DIA and EAO and increased during early postnatal development before declining to adult levels. FR did not significantly correlate with SDH activity (r2 = 0.01) but did relate to the MHC phenotype as indexed by the ratio of adult MHC isoform content (slow + IIa + IIx + IIb) to developmental MHC isoform content (slow + neonatal; r2 = 0.88, P < 0.01). Stepwise regression revealed that neonatal MHC expression alone accounted for 60% of the developmental variance in FR. The correlation between FR and MHC phenotype was improved if SDH was also considered, i.e., the ratio of SDH to MHC phenotype (r2 = 0.99, P < 0.01). We conclude that FR of respiratory muscle during development relates to a balance between the energetic demands of the muscle contractile proteins as reflected by MHC isoform composition and its oxidative capacity with MHC phenotype alone exerting a strong predictive effect on FR. PMID- 8226550 TI - Sulfide toxicity: mechanical ventilation and hypotension determine survival rate and brain necrosis. AB - Occupational exposure to hydrogen sulfide is one of the leading causes of sudden death in the workplace, especially in the oil and gas industry. High-dose exposure causes immediate neurogenic apnea and death; lower doses cause "knockdown" (transient loss of consciousness, with apnea). Because permanent neurological sequelae have been reported, we sought to determine whether sulfide can directly kill central nervous system neurons. Ventilated and unventilated rats were studied to allow administration of higher doses of sulfide and to facilitate physiological monitoring. It was extremely difficult to produce cerebral necrosis with sulfide. Only one of eight surviving unventilated rats given high-dose sulfide (a dose that was lethal in > or = 50% of animals) showed cerebral necrosis. Mechanical ventilation shifted the dose that was lethal in 50% of the animals to 190 mg/kg from 94 mg/kg in the unventilated rats. Sulfide was found to potently depress blood pressure. Cerebral necrosis was absent in the ventilated rats (n = 11), except in one rat that showed profound and sustained hypotension to < or = 35 Torr. Electroencephalogram activity ceased during exposure but recovered when the animals regained consciousness. We conclude that very-high-dose sulfide is incapable of producing cerebral necrosis by a direct histotoxic effect. PMID- 8226551 TI - Exercise training alters endothelium-dependent vasoreactivity of rat abdominal aorta. AB - We tested the hypothesis that adaptations in peripheral arterial vasoreactivity are induced by exercise training. Male rats were trained to run on a treadmill at 30 m/min (15 degrees incline) for 1 h/day 5 days/wk for 10-12 wk. Efficacy was indicated by a 51% increase (P < 0.05) in citrate synthase activity in soleus muscle of exercise-trained (ET) rats compared with that of sedentary (SED) control rats. Responses to vasoactive compounds were examined in vitro using rings of abdominal aorta. Maximal isometric contractile tension evoked by KCl, norepinephrine (NE), and phenylephrine were not different between groups; sensitivity to phenylephrine was also not different between groups. However, sensitivity was lower for both KCl and NE in vessels from ET animals. Endothelium removal did not influence KCl sensitivity but did abolish the difference in NE sensitivity of vessel segments between ET and SED animals. Maximal vasodilator responses induced by acetylcholine (ACh; NE or prostaglandin F2 alpha preconstriction) were greater in vessel rings from ET rats. However, dilatory responses by sodium nitroprusside (NE or prostaglandin F2 alpha preconstriction) and forskolin (NE preconstriction) were not different between groups, indicating that the augmented ACh-induced dilatory response resulted from an adaptation of the endothelium. Blockade of nitric oxide synthase activity diminished ACh induced vasodilation by 79 and 100% in SED and ET rats, respectively. These results indicate that training alters vasomotor function in rat abdominal aortas through adaptations of both endothelium and smooth muscle. PMID- 8226552 TI - Effect of diaphragmatic fatigue on control of respiratory muscles and ventilation during CO2 rebreathing. AB - We studied the influence of diaphragmatic fatigue on the control of ventilation and respiratory muscle contribution to pressure swings in six normal seated subjects. CO2 was rebreathed before and after diaphragmatic fatigue induced by breathing against an inspiratory resistance requiring 60% maximal transdiaphragmatic pressure with each breath until exhaustion. After diaphragmatic fatigue for a given level of end-tidal PCO2, we found that tidal volume, breathing frequency, minute ventilation, duty cycle, and mean inspiratory flow did not change; esophageal pressure swings were the same, but gastric and transdiaphragmatic pressure swings were decreased; and the slope of the transpulmonary pressure-gastric pressure relationship determined at zero flow points at end expiration and end inspiration was increased. End-expiratory transpulmonary pressure progressively decreased and end-expiratory gastric pressure progressively increased with increasing end-tidal PCO2 by the same magnitude before and after diaphragmatic fatigue. We conclude that diaphragmatic fatigue induces proportionately greater contributions of inspiratory rib cage muscles than of the diaphragm, which results in the preservation of ventilatory response to CO2 despite impaired diaphragmatic contractility. PMID- 8226553 TI - Effect of global inspiratory muscle fatigue on ventilatory and respiratory muscle responses to CO2. AB - We evaluated the effect of global inspiratory muscle fatigue on ventilation and respiratory muscle control during CO2 rebreathing in normal subjects. Fatigue was induced by breathing against a high inspiratory resistance until exhaustion. CO2 response curves were measured before and after fatigue. During CO2 rebreathing, global fatigue caused a decreased tidal volume (VT) and an increased breathing frequency but did not change minute ventilation, duty cycle, or mean inspiratory flow. Both esophageal and transdiaphragmatic pressure swings were significantly reduced after global fatigue, suggesting decreased contribution of both rib cage muscles and diaphragm to breathing. End-expiratory transpulmonary pressure for a given CO2 was lower after fatigue, indicating an additional decrease in end expiratory lung volume due to expiratory muscle recruitment, which leads to a greater initial portion of inspiration being passive. This, combined with the reduction in VT, decreased the fraction of VT attributable to inspiratory muscle contribution; therefore the inspiratory muscle elastic work and power per breath were significantly reduced. We conclude that respiratory control mechanisms are plastic and that the respiratory centers alter their output in a manner appropriate to the contractile state of the respiratory muscles to conserve the ventilatory response to CO2. PMID- 8226554 TI - Carotid chemoreceptors and respiratory adaptations to dead space loading during incremental exercise. AB - Dead space (VD) loading has been shown to cause an increase in tidal volume and a decrease in respiratory frequency at moderate to high levels of ventilation (VI) during exercise (J. Appl. Physiol. 70: 55-62, 1991). This study examined the role of carotid chemoreceptors (CC) in the breathing pattern response to added VD during maximal incremental exercise; we used hyperoxia to silence the CC. Nine healthy subjects exercised on a bicycle ergometer on 4 different days while inspiring air with VD (AVD) and without VD [air control (AC)] and while inspiring 100% O2 with VD (O2VD) and without VD (O2C). Equipment resistance for VD and control studies was identical, and the exercise tests were done in a randomized order. At a matched level of VI equivalent to 75% VI at the end of the AC experiments (102 l/min), the breathing pattern in the AVD and O2VD tests was significantly deeper and slower (P < 0.05) than that in the AC and O2C tests. The difference in tidal volume between AVD and AC tests (delta = 0.26 +/- 0.16 liter) was not significantly different from that between O2VD and O2C tests (delta = 0.23 +/- 0.23 liter). The breathing pattern was the same in the AC and O2C tests. It is concluded that the altered breathing pattern with VD loading is not mediated by the CC. PMID- 8226555 TI - Effect of asthma and ventilatory loading on arterial PCO2 of humans during submaximal exercise. AB - In humans, attenuating carotid chemoreceptor activity by hyperoxia does not alter arterial PCO2 (PaCO2) during submaximal exercise, yet a transient hypercapnia occurs in carotid chemoreceptor-resected (CBR) asthmatic subjects during submaximal exercise. We hypothesized that this difference was due to asthma and not CBR causing the abnormal response. Accordingly, we determined the temporal pattern of PaCO2 during mild and moderate exercise in chemoreceptor-intact asthmatic (n = 10) and nonasthmatic subjects (n = 10). We also hypothesized that hyperoxia alters PaCO2 during exercise if exercise already has disrupted PaCO2 homeostasis. Accordingly, we studied, during exercise, asthmatic subjects while hyperoxic; nonasthmatic subjects during loaded breathing of room air, which increased PaCO2; and nonasthmatic subjects during loaded breathing while hyperoxic. While breathing room air, neither asthmatic nor nonasthmatic subjects maintained arterial isocapnia during exercise. An increase in PaCO2 between rest and exercise and between mild exercise and 1st min of moderate exercise was greater in asthmatic than in nonasthmatic subjects (P < 0.05). In six asthmatic subjects that were hypercapnic breathing room air during exercise, hypercapnia was accentuated by hyperoxia. The ventilatory load in nonasthmatic subjects resulted in a work load-dependent hypercapnia (P < 0.01) accentuated (P < 0.01) by hyperoxia. We conclude that normally in humans the carotid chemoreceptors contribute minimally to the hyperpnea of submaximal exercise. However, when PaCO2 is increased from resting values during exercise, then the chemoreceptors serve to augment ventilation and thereby minimize the hypercapnia. PMID- 8226556 TI - Hypoglossal and phrenic nerve responses to carotid baroreceptor stimulation. AB - We examined the relationship between hypoglossal and phrenic nerve activities and carotid sinus pressure. In 12 adult cats that were decerebrate, vagotomized, paralyzed, and mechanically ventilated, we isolated the left carotid sinus for perfusion and denervated the right carotid sinus. Mean arterial blood pressure was maintained at 90-100 mmHg using a low resistance-reservoir containing saline and connected to the abdominal aorta. Constant pressure was applied to the carotid sinus region. We found that increased carotid sinus pressure immediately inhibited inspiratory-synchronous (phasic) hypoglossal nerve activity and that there was a direct inverse relationship between phasic hypoglossal activity and carotid sinus pressure up to a carotid pressure of 285 mmHg. Increased carotid sinus pressure had no effect on tonic hypoglossal nerve activity and only slightly inhibited phrenic nerve activity. Cutting the left carotid sinus nerve abolished this response. We also applied pressure pulses to the carotid sinus at discrete times during the phrenic cycle. We found that baroreceptor inhibition of phasic hypoglossal nerve activity was gated during the phrenic cycle: maximum inhibition occurred when the pulse was applied in late expiration. We conclude that carotid baroreceptor stimulation preferentially inhibits inspiratory synchronous hypoglossal nerve activity and that this afferent information traveling in the carotid sinus nerve is gated by the respiratory control center. PMID- 8226557 TI - Role of tachykinins in ozone-induced acute lung injury in guinea pigs. AB - To examine the hypothesis that the acute reversible changes caused by ozone (O3) exposure are mediated by tachykinin release, guinea pigs were depleted of tachykinins by use of repeated capsaicin (CAP) injections before O3 exposure in an attempt to prevent O3-induced functional changes. Unexpectedly, CAP pretreatment caused divergent results in the functional responses to O3. Ventilatory measurements obtained from CAP-pretreated O3-exposed (CAP-O3) animals were exacerbated rather than diminished compared with the effects of O3 alone. Similarly, lavage fluid protein accumulation was enhanced in the CAP-O3 group compared with the O3-exposed group. In better agreement with our initial hypothesis, the CAP-O3 group was less responsive than the O3-exposed animals to histamine aerosol challenge. Additionally, Evans blue dye accumulation, a hallmark of tachykinin release, was increased in O3-exposed animals and was partially blocked in the CAP-O3 group. These data suggest that tachykinin containing sensory fibers are unlikely to mediate the acute effects of O3 exposure on tidal breathing and lavage fluid protein accumulation but may play a role in causing post-O3 airway hyperreactivity and protein extravasation into the trachea. PMID- 8226558 TI - Determination of human skeletal muscle buffer value by homogenate technique: methods of measurement. AB - The human muscle buffer value (beta) is most frequently determined by either fixed acid titration of a homogenate ["in vitro" beta (beta vit)] or measurement of the change in lactate concentration (delta [La]) relative to the change in muscle homogenate pH after high-intensity exercise ["in vitro" beta = - delta [La]/delta pH (beta viv)]. We sought to compare beta viv, determined after isometric and dynamic exercise to exhaustion (approximately 60 s), with beta vit. Resting (R) and postexercise (E) biopsy samples were taken from vastus lateralis muscles of 43 human volunteers. Freeze-dried muscle was homogenized (30 mg/ml) in NaF (0.01 M) for the measurement of muscle pH (R and E). beta vit was determined by HCl (0.01 M) titration of the homogenate over the pH range 7.1-6.5. Muscle lactate was measured by enzymatic assay. There was no significant difference between beta viv determined after isometric (n = 35) or dynamic (n = 8) exercise to fatigue (170 vs. 168 mmol H+.kg dry muscle mass-1.pH-1, respectively; P > 0.05). Values for beta vit in the corresponding muscle samples (R) were approximately 7-8% lower (156 +/- 25 vs. 157 +/- 18 mmol H+.kg dry muscle mass 1.pH-1, respectively). There was no significant difference (P = 0.278) between the measured decline in muscle homogenate pH after exercise and the reduction in pH predicted from beta vit and delta [La], indirectly confirming the lack of any significant difference between beta viv and beta vit.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226559 TI - Method for computing the oxidation of two 13C-substrates ingested simultaneously during exercise. AB - This study presents a method for computing the respective amounts of two simultaneously ingested exogenous substrates (A and B) that are oxidized during a period of prolonged exercise by use of 13C labeling. This method is based on the observation that the total volume of 13CO2 produced (V13CO2tot) is the sum of 1) V13CO2 arising from the oxidation of endogenous substrates (V13CO2endo), 2) V13CO2 arising from the oxidation of substrate A (V13CO2A), and 3) V13CO2 arising from the oxidation of substrate B (V13CO2B). The equation, V13CO2tot = V13CO2endo+V13CO2A+V13CO2B, with three unknowns, can be solved from the results of three experiments conducted under the same conditions but with at least two values for the isotopic composition of A and B. This method has been used on five healthy male subjects to compute the amounts of glucose and fructose oxidized when a mixture of 15 g of glucose and 15 g of fructose is ingested (in 300 ml of water) over 60 min of cycle ergometer exercise at 65% of maximal O2 uptake. Results from three experiments indicated that 9.8 +/- 3.1 and 5.7 +/- 2.1 g of glucose and fructose, respectively, were oxidized. The total amount of exogenous carbohydrates oxidized (15.5 +/- 4.3 g) is in agreement with the oxidation rates of exogenous glucose computed in similar conditions when 30 g of glucose were ingested (13 g; Peronnet et al. Med. Sci. Sports Exercise 25: 297-302, 1993). The difference between the oxidation rates of exogenous glucose and fructose is also in line with data from the literature. PMID- 8226560 TI - HEPES buffer perfusate alters rabbit lung endothelial permeability. AB - N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) has been shown to cause changes in cultured endothelial cells and smooth muscle function at concentrations from 5 to 25 mM. To determine whether HEPES also affects vascular permeability, the effects of two buffers, HEPES and phosphate, were compared in isolated perfused rabbit lungs. Hemodynamic parameters and vascular protein permeability-surface area products (PS) were measured after perfusion with the buffers. Endothelial permeability was measured for an anionic and a cationic albumin to assess the charge effects of the zwitterion buffer. With HEPES, there were no changes in vascular pressure or resistance but permeability was affected. Cationic albumin permeability increased with 12 mM HEPES (8.7phosphate-->30(12) mM HEPES x ml.min-1.g dry lung-1 x 10(-2)) as did the anionic albumin PS (2.7phosphate-->3.52(12) mM HEPES). The cationic PS returned to baseline (8.1(60) mM HEPES) at 60 mM HEPES, but the anionic PS did not change from the 12 mM HEPES (4.01(60) mM HEPES). In summary, we find that HEPES is not innocuous. Although hemodynamic parameters did not change, endothelial permeability was increased when HEPES was used at normal concentrations. Therefore, HEPES should be used with caution as a physiological buffer in perfused organ systems. PMID- 8226561 TI - Influence of active muscle size on sympathetic nerve discharge during isometric contractions in humans. AB - We tested the hypothesis that sympathetic nerve discharge to nonactive skeletal muscle (MSNA) is influenced by active muscle size during isometric contractions performed at a constant submaximal force in humans. In six subjects, MSNA (peroneal microneurography), arterial pressure, heart rate, and ratings of perceived effort were recorded before (resting control) and during isometric contractions of either a small hand muscle [1st dorsal interosseus (FDI); 2 trials (FDI1 and FDI2)] or the fore- and upper arm muscles (handgrip; 1 trial) sustained to the point of exhaustion and normalized to endurance time (te). There were no differences in resting control levels. MSNA, heart rate, arterial pressure, and perceived effort all increased similarly during the two FDI contractions. During handgrip, the rates of rise and peak increases MSNA, heart rate, and arterial pressure were much greater than during FDI (all P < 0.05), but peak levels of perceived effort were not different. These findings indicate that during voluntary isometric efforts sustained to the same performance and perceptual end points the rates of increase and final levels of MSNA, heart rate, and arterial pressure are greater during contraction of a larger muscle mass and that these responses appear to be similar during successive trials of a task when normalized to te. Thus, active muscle size can be an important factor in the regulation of sympathetic nervous system discharge and cardiovascular function during isometric muscle activity in the human. PMID- 8226562 TI - Cardiovascular responses in black and white males during exercise? PMID- 8226563 TI - Management of immunocompromised patients with evidence of an invasive mycosis. AB - Invasive fungal infections have emerged as major causes of morbidity and mortality in immunocompromised patients, particularly granulocytopenic and HIV infected hosts. Diagnosis and treatment of these infections is determined by the type of fungus and the sites of infection. While Candida spp. and Aspergillus spp. are the most common causes of fungal infection in granulocytopenic patients, less common pathogens are emerging as new challenges. This article summarizes current approaches used in the National Cancer Institute's Pediatric Branch for management of invasive fungal infections. PMID- 8226564 TI - Critical appraisal of immunization strategies for prevention of infection in the compromised host. AB - Chemotherapy and/or bone marrow transplantation clearly places individuals at increased risk for many vaccine-preventable diseases. Both active and passive immunization strategies have been considered for protecting immunocompromised patients. Many compromised hosts are capable of mounting protective responses to some protein toxoids, polysaccharide-conjugate, and viral vaccines. Effective, safe, and practical dosage schedules are evolving for individual vaccines and specific patient populations. PMID- 8226565 TI - Critical appraisal of antimicrobials for prevention of infections in immunocompromised hosts. AB - Infections remain a common cause of morbidity and mortality among patients receiving antineoplastic chemotherapy. Over the past 30 years a number of techniques have been evaluated in an attempt to reduce the incidence of bacterial, fungal, and viral infections. Protective isolation, oral nonabsorbable antibiotics, and selective decontamination of the gastrointestinal tract have all been extensively evaluated. More recently, newer antimicrobial agents have been employed, such as quinolones, imidazoles, and recombinant monoclonal agents for modulation of the immune cascade. A critical review of the advantages and disadvantages of each of these methodologies is presented. PMID- 8226566 TI - Approach to treatment of the febrile cancer patient with low-risk neutropenia. AB - All cancer patients who develop fever during a period of neutropenia are not at high risk of developing serious infection. Following hospitalization for empiric broad spectrum antibiotic therapy, certain low-risk patients can be identified and safely be discharged "early", prior to recovery of the absolute neutrophil count to 500 cells per mm3. The medical, psychological, and financial benefits of early hospital discharge are immense. Careful examination for serial increases in components of the peripheral blood count herald marrow recovery and help identify the low-risk patient. It may also be possible to identify "low-risk" neutropenic patients upon development of fever in order to explore alternative management strategies (for example, outpatient oral or intravenous antibiotics) rather than routine hospitalization. PMID- 8226567 TI - Management of the cancer patient with fever and prolonged neutropenia. AB - Cancer patients who suffer prolonged durations of fever and neutropenia are at high risk for developing serious infections. This article reviews the initial management of these fevers, subsequent modifications of the initial empirical regimens (both empirical changes in the antibiotic regimen and those changes directed at specific sites of infection that may become apparent during the course of prolonged neutropenia), and the potential impact of hematopoietic cytokines on future management of prolonged fever and neutropenia. PMID- 8226568 TI - Special considerations for the patient undergoing allogeneic or autologous bone marrow transplantation. AB - Improvements in the diagnosis, treatment, and prevention of infectious complications of bone marrow transplantation over the past two decades have markedly reduced the morbidity and mortality of this procedure. We are now able to begin early empiric antibiotic coverage with less toxic, but equally effective, antibacterial agents. Once believed to be uniformly fatal, complications such as CMV pneumonia are now considered treatable in at least half the cases with a combination of intravenous immunoglobulin and ganciclovir. Although probably the most controversial, prophylactic therapy has improved the outcome of patients undergoing bone marrow transplantation. The appropriate setting, agents to use, dose, and dose intervals will require further study in coming years. In the introduction to this article, we attempted to outline what is known about the immunobiology of bone marrow transplantation. A clear understanding of this process helps us recognize and anticipate the infectious complications encountered in this population of patients. It may also allow clinicians to focus more on immune augmentation as a means of prevention, as has been attempted with the newly available cytokines and the use of intravenous immunoglobulin infusions. Despite improvements in diagnosis, treatment, and prevention, infectious complications remain the leading cause of morbidity and mortality in the patient undergoing bone marrow transplantation. Future studies are required in this area to build on the successes of the last two decades. PMID- 8226569 TI - Tobaccics--a doomed generation. PMID- 8226570 TI - Endoscopic sinus surgery. A 300 case review. AB - The introduction of ESS (endoscopic sinus surgery) has changed and vastly improved the potential of sinus surgery. It is now possible to visualize obstruction in the natural sinus ostia and remove diseased tissue, thereby promoting restoration of normal function. A series of 300 consecutively performed cases is reviewed; results and complications are reported. Findings include a success rate of 94%, with complications in less than 1%. ESS offers a major advancement in treatment of chronic sinusitis. PMID- 8226571 TI - I thought I saw that report. PMID- 8226572 TI - Doctors respond with signs about Medicare assignment. PMID- 8226573 TI - Despite intensive efforts, egg-related salmonellosis outbreaks continue. PMID- 8226574 TI - Radiological case of the month. Dermoid cyst. PMID- 8226575 TI - Arkansas native provides AIDS information on compact discs. PMID- 8226576 TI - Transjugular intrahepatic portosystemic shunt (TIPS) in the management of variceal hemorrhage. AB - Transjugular intrahepatic portosystemic shunt placement is a new non-operative therapeutic procedure used by interventional radiologists to manage patients with variceal hemorrhage who are unresponsive to medical treatment. This procedure offers an alternative to surgical shunt procedures with a much lower morbidity and mortality and has a high success rate in controlling bleeding. The early experience at St. Vincent Infirmary Medical Center is discussed. PMID- 8226577 TI - The pediatric anesthesiologist as consultant to the pediatrician. AB - Physicians with training in pediatrics and anesthesiology may be useful as consultants to primary care physicians in the community. Pediatricians or family practitioners may encounter patients with chronic or intractable pain that requires multidisciplinary therapy. Local anesthesiologists may encounter children with complex medical disease or unfamiliar syndromes. Information and consultation can be obtained by contacting the Division of Pediatric Anesthesia or the Pain Management Program at Arkansas Children's Hospital. This discussion emphasizes the knowledge-base associated with the specialty and suggests scenarios in which this knowledge and training might be used. PMID- 8226578 TI - Tuberculosis and HIV infection. PMID- 8226579 TI - Radiological case of the month. Free intraperitoneal air. PMID- 8226580 TI - Can standard measures identify subclinical markers of autism? AB - This study compared the executive function and theory-of-mind abilities of siblings of autistic individuals to those of siblings of learning-disabled controls. Three different analyses of the dependent measures provided convergent support for a potential subclinical marker in the executive function domain. No group differences in theory-of-mind abilities were found. However, power analyses revealed that the measures employed in this study, which are typically used with autistic individuals, were not sufficiently sensitive to detect any group differences that might exist in "unaffected" family members. Suggestions for future research are provided, including the need to develop more sensitive tasks that produce larger effects and measure more elementary cognitive operations. PMID- 8226581 TI - A neural network approach to the classification of autism. AB - A nonlinear pattern recognition system, neural network technology, was explored for its utility in assisting in the classification of autism. It was compared with a more traditional approach, simultaneous and stepwise linear discriminant analyses, in terms of the ability of each methodology to both classify and predict persons as having autism or mental retardation based on information obtained from a new structured parent interview: the Autistic Behavior Interview. The neural network methodology was superior to discriminant function analysis both in its ability to classify groups (92 vs. 85%) and to generalize to new cases that were not part of the training sample (92 vs. 82%). Interrater and test retest reliabilities and measures of internal consistency were satisfactory for most of the subscales in the Autistic Behavior Interview. The implications of neural network technology for diagnosis, in general, and for understanding of possible core deficits in autism are discussed. PMID- 8226582 TI - Integrated play groups: a model for promoting the social and cognitive dimensions of play in children with autism. AB - This investigation provides a description of a multifaceted model to promote peer play, and an evaluation of its impact on the social and cognitive dimensions of play in three children with autism. A combination of quantitative and qualitative methodologies were used to evaluate the effectiveness of the model application. A multiple-probe design across participants demonstrated (a) decreases in isolate play and collateral gains in more social forms of play, and (b) decreases in stereotyped object play and collateral gains in functional object play. While no symbolic play was observed in any of the participants during baseline, two participants demonstrated symbolic play in the final condition. Generalization and social validation measures indicated (a) advances in play behaviors were not limited to the play groups but observed in other contexts, and (b) were accompanied by language gains. Implications are discussed in terms of preferred service delivery models as well as of the importance of social interaction for the development of play and language. PMID- 8226583 TI - Fenfluramine and methylphenidate in children with mental retardation and attention deficit hyperactivity disorder: laboratory effects. AB - Twenty-eight children took part in a double-blind, placebo-controlled, crossover study of fenfluramine and methylphenidate. Fenfluramine dosage was gradually increased to a standardized dose of 1.5 mg/kg per day, whereas methylphenidate was given in doses of 0.4 mg/kg per day. The children were assessed on laboratory tests of selective and sustained attention, visual matching, and color matching, during which seat activity was monitored automatically. Results showed fenfluramine to be superior to placebo on the memory task, whereas methylphenidate reduced commission errors on a continuous performance test. Methylphenidate caused shorter response times, and fenfluramine caused increases, on two of the tests. Examiner behavior ratings indicated significant improvements with both drugs on the domains of attention, activity level, and mood. These findings, together with those from a companion clinical study, suggest that the drugs may have contrasting mechanisms of action, but both appear to have useful clinical effects in these children. PMID- 8226584 TI - Facilitated communication: a failure to replicate the phenomenon. AB - Twenty-one subjects participated in a study of Biklen's and Crossley's hypothesis that persons with autism show unexpected literacy and improved communication ability through the process of facilitated communication (FC). Repeated measures of literacy were conducted at (a) a baseline test of communicative ability before FC; (b) a pretest with facilitation; and (c) a posttest with facilitation after 20 hours of training. At both the pretest and posttest, the facilitators were screened from hearing or seeing the questions or pictorial stimuli. Although some facilitators reported newfound communicative abilities during training sessions, no client showed unexpected literacy or communicative abilities when tested via the facilitator screening procedure, even after 20 hours of training. Separate analyses indicated that some facilitators influenced the communicative output of their clients. PMID- 8226585 TI - Brief report: evaluation of eight case studies of facilitated communication. PMID- 8226586 TI - Brief report: facilitator-suggested conversational evaluation of facilitated communication. PMID- 8226587 TI - Brief report: facilitated communication evaluation procedure accepted in a court case. PMID- 8226588 TI - Brief report: an empirical evaluation of spelling boards as a means of communication for the multihandicapped. PMID- 8226589 TI - Commentary on the possible role of oxytocin in autism. PMID- 8226590 TI - The three-dimensional modeling ability of a boy with autism. PMID- 8226591 TI - Diagnosis by DSM-III-R versus ICD-10 criteria. PMID- 8226592 TI - Renal transplantation in India-problems and prospects. PMID- 8226593 TI - Live related renal transplantation for end stage diabetic nephropathy. AB - We report our results with live related renal transplantation in 43 diabetics, most of whom were non-insulin dependent, with end stage renal disease. The overall one year patient survival was 72.1% and graft survival was 65.1%. The use of Cyclosporine was associated with a significant improvement in the one year patient and graft survival (92.3% and 84.6% respectively). The most important cause of mortality was infection. Live related renal transplantation with Cyclosporine as immunosuppression is advisable for the uremic diabetic. PMID- 8226594 TI - Clinico-therapeutic profile of pyogenic liver abscess. AB - Nineteen consecutive patients of pyogenic liver abscess (13 males, 6 females, mean age 45.2 +/- 6, 5 years) were studied over a period of 5 years for the clinical profile and therapeutic efficacy of percutaneous aspiration. Majority of them presented with spiking fever (94.7%), pain over right upper quadrant of abdomen (53.8%) and often with prostration and shock (31.6%). The onset has been rather acute in patients with multiple abscesses (7 cases). Ascites (10.5%), clubbing of fingers (15.8%) and splenomegaly (10.5%) were observed in cases with long duration of illness. There has been the polymorphonuclear leucocytosis (89.5%) and mild to moderate anaemia (52.6%). Jaundice (42.1%) was usually mild degree (serum bilirubin 4.6 +/- 2.4 mg/dl). The serum transaminases and alkaline phosphatase were raised in 94.9% of cases. Ultrasonography revealed predominantly hypoechoic (54.1%) areas with frequent distal acoustic enhancement and internal echoes (21.6%). Seven patients had multiple abscesses with 25 lesions (size 5.2 +/- 4.6 cms) situated mainly over antero-inferior segment of the right lobe (45.9%) of the liver. The blood and pus cultures demonstrated the micro-organisms (positive in 63.9% and 86.6% respectively) predominantly of bowel flora including facultative gram negative rods and anaerobes. Mortality was 26.5% and pleuro pulmonary complications were the commonest (26.4%) one. 15 cases were treated by percutaneous aspiration, proper antibiotic and metronidazole with encouraging results (only two deaths). Complete resolution of abscess took place in 14.2 +/- 1.2 weeks (range 6 weeks to 7.5 months). It is emphasized that percutaneous needle aspiration is useful both for diagnosis and treatment of pyogenic liver abscess. PMID- 8226595 TI - Significance of minor electrocardiographic repolarisation changes in asymptomatic subjects. AB - 3000 routine ECGs were screened during a 2 year period in Armed Forces personnel reporting for annual medical examination. 90 asymptomatic men with minor ST segment and T wave changes at rest were assessed by exercise ECG. 33 responded abnormally. They were subjected to coronary arteriography. 14 subjects were confirmed to have coronary artery disease (CAD). 19 subjects had a false positive response and assuming a false negative response of 2%, a sensitivity of 87.5%, specificity of 74.3% and a positive predictive value of 42.4% is obtained for exercise ECG. A positive predictive value of 15.5% is obtained for the resting ECG. PMID- 8226596 TI - Assessment of exercise tolerance in chronic bronchitis and interstitial lung disease. AB - Cardiopulmonary exercise testing has a definitive place in objective evaluation of the subjective sensation of breathlessness. In the present study 21 patients with COPD, 10 with ILD and 17 normals were subjected to stage 1 exercise testing and correlations were sought between Vo2 and work, FVC, FeV1, Ve, respiratory rate and tidal volume and between VCO2 and work. In COPD and ILD the Vo2 correlated with work, Fev1, Fvc and Ve whereas in normals it correlated with the VE but not with the FeV1 and FVC. In COPD VO2 correlated with TV though this was not the case in ILD. Thus stage 1 exercise testing can be a useful additional method to assess the disability in COPD and ILD although differentiation between these two respiratory diseases on basis of exercise testing alone may not be possible. PMID- 8226597 TI - Nuclear imaging study of nifedipine induced modulations of central haemodynamics in severe hypertension with and without coronary artery disease. AB - Acute effects of Nifedipine, a calcium channel blocking agent (CCBA), on central haemodynamics in patients with severe hypertension and angiographically proved coronary artery disease (CAD) has been studied using Nuclear Ventriculography (MUGA). While peripheral vasodilatation leads to significant reduction of systemic blood pressure in all severe hypertensive patients (p < 0.0005 and p < 0.0001), the central haemodynamics did not improve significantly in patients without CAD (p = NS). However, in hypertensive subset of CAD, central haemodynamics significantly improves with rise of global ejection fraction (p < 0.001) and improvement of wall motion abnormalities (p < 0.001). PMID- 8226598 TI - Renal functions in obstructive jaundice: a pre and post operative assessment. AB - Surgery in patients with surgical obstructive jaundice is known to be associated with increased risk of post-operative acute renal failure. A prospective study was carried out to evaluate the renal function in patients with obstructive jaundice. Renal functions of thirty two patients with jaundice secondary to mechanical obstruction of the biliary tract were evaluated pre-operatively and 7 days after surgical biliary decompression. Although no significant difference was seen in the mean values of pre and post operative renal function variables, two patients had overt renal failure, one with pre-operative cholangitis and acute tubular necrosis and another with carcinoma of the pancreas and postoperative acute renal failure. The overall satisfactory outcome in our obstructive jaundice patients may be related to pre-operative and intraoperative preparation with intravenous fluids and mannitol. It is concluded that patients with obstructive jaundice can be satisfactorily treated with special preoperative care including good hydration and mannitol therapy during anaesthesia and surgery. PMID- 8226599 TI - Study of cerebrospinal fluid in Salmonella typhi meningitis. AB - The present study was designed to study the cerebrospinal fluid of Salmonella typhi meningitis cases. All the 12 cases (100%) showed increase in the lymphocyte count (range 6-24 cells/cmm, mean 15.40 cells/cmm) and increase in protein concentration (range 80-125 mg.Dl, mean 86.25 mg/dl), while glucose concentration remained normal (range 56-80 mg/dl, mean 64.25 mg/dl). These changes in CSF may resemble viral meningitis, from which Salmonella typhi meningitis can only be differentiated by culture of CSF. PMID- 8226600 TI - Engima of therapeutic failure in visceral leishmaniasis. PMID- 8226601 TI - Cardiomyoplasty--current status. AB - Dynamic cardiomyoplasty is a new surgical technique that uses transformed (electrostimulated) skeletal muscle to reinforce or replace damaged myocardium. The clinical benefits of cardiomyoplasty have not been as dramatic as its evolution. However, with the limitations of cardiac transplantation and with further developments in cardiomyoplasty, this technique is likely to be established as a viable alternative in selected patients with refractory cardiac failure. Basic research and animal studies suggest that there could be a possible role of cardiomyoplasty in certain pediatric cardiac disorders. PMID- 8226602 TI - Cytology in the diagnosis of thyroid lesions--a review. AB - Fine needle aspiration cytology is today one of the first line investigations for palpable and deep seated masses. Non aspiration fine needle cytology is another recently described cytologic technique. The thyroid gland is one of the common head and neck structures to be subjected to cytologic sampling. Cytology has proved to be a highly sensitive and specific investigative modality in solitary cold thyroid nodules on the basis of which the therapeutic protocol is decided. By providing an accurate morphologic diagnosis in most of the neoplastic lesions, cytology enables pre-operative planning of the extent of surgery. It is also an excellent method for the diagnosis and monitoring of patients harboring inflammatory or autoimmune thyroid lesions. PMID- 8226603 TI - Deceleration--dependent left bundle branch block in rheumatic carditis. PMID- 8226604 TI - Meige's syndrome. AB - A case of Meige's syndrome is reported who presented with blepharospasm and oromandibular dystonia along with dislocation of jaw, due to severe dystonia. Such severe form of dystonia are very rare. PMID- 8226605 TI - Toxic strep syndrome. AB - Group A beta-hemolytic streptococci are known to produce infections with protean manifestations. We report here a case of toxic streptococcal syndrome in a patient who presented with a multisystemic disease. Serological and bacteriological evidence strongly suggested an infection by Group A beta hemolytic streptococci. PMID- 8226606 TI - Acute megaloblastic reaction in hereditary spherocytosis. PMID- 8226607 TI - Unilateral ataxia following herpes zoster of spinal C4 segment. AB - Focal lesions of central nervous system are extremely rare following cutaneous herpes zoster. A 55 year old male developed cerebellar speech, right sided ataxia and intention tremor, three weeks after herpes zoster of right spinal C4 segment. Clinical examination and investigations confirmed a focal vascular lesion in the midbrain suggestive of granulomatous angiitis which can cause focal neurological defect after herpes zoster. PMID- 8226608 TI - Eales' disease with progressive spastic paraparesis. AB - Eales disease is a disease of unknown aetiology characterised by ophthalmic changes and often neurological involvement as in multiple sclerosis. We are reporting a case of Eales' disease in a 27 yr old male with chronic progressive non compressive motor myelopathy. PMID- 8226609 TI - Myocardial decompensation in heroin addicts. PMID- 8226610 TI - Changing trends in the treatment of enteric fever. PMID- 8226611 TI - Clicking rib. PMID- 8226612 TI - Aluminium phosphide poisoning. PMID- 8226613 TI - Is the magic of fluoroquinolones waning? PMID- 8226614 TI - Nitrogenase metalloclusters: structures, organization, and synthesis. PMID- 8226615 TI - Purification and characterization of maleylacetate reductase from Alcaligenes eutrophus JMP134(pJP4). AB - Maleylacetate reductase (EC 1.3.1.32) plays a major role in the degradation of chloroaromatic compounds by channeling maleylacetate and some of its substituted derivatives into the 3-oxoadipate pathway. The enzyme was purified to apparent homogeneity from an extract of 2,4-dichlorophenoxyacetate (2,4-D)-grown cells of Alcaligenes eutrophus JMP134. Maleylacetate reductase appears to be a dimer of two identical subunits of 35 kDa. The pI was determined to be at pH 5.4. There was no indication of a flavin prosthetic group. The enzyme was inactivated by p chloromercuribenzoate but not by EDTA, 1,10-phenanthroline, or dithiothreitol. Maleylacetate and 2-chloromaleylacetate were converted with similar efficiencies (with NADH as cosubstrate, Km = 31 microM for each substrate and kcat = 8,785 and 7,280/min, respectively). NADH was preferred to NADPH as the cosubstrate. Upon reduction of 2-chloramaleylacetate by the purified enzyme, chloride was liberated and the resulting maleylacetate was further reduced by a second NADH. These results and the kinetic parameters suggest that the maleylacetate reductase is sufficient to channel the 2,4-D degradation intermediate 2-chloromaleylacetate into the 3-oxoadipate pathway. In a data base search the NH2-terminal sequence of maleylacetate reductase was found to be most similar to that of TfdF, a pJP4 encoded protein of as-yet-unknown function in 2,4-D degradation. PMID- 8226616 TI - Hypersensitive sites in the 5' promoter region of nit-3, a highly regulated structural gene of Neurospora crassa. AB - The nit-3 gene of the filamentous fungus Neurospora crassa encodes nitrate reductase, the enzyme which catalyzes the first step in nitrate assimilation. The nit-3 gene is subject to a high degree of regulation by metabolic inducers and repressors, and its expression requires two distinct trans-acting regulatory proteins. Hypersensitive sites in the 5' DNA sequence upstream of the nit-3 gene were mapped with the use of three different nucleases as molecular probes. Six hypersensitive sites, three of which are very strong, were detected at essentially identical positions by all three nucleases. The hypersensitive sites appear to develop in a constitutive fashion and are present under conditions in which the nit-3 structural gene is expressed but also when this gene is inactive, although these sites are considerably less prominent in cells subjected to nitrogen catabolite repression. The presence of the hypersensitive sites appears to depend upon both the positively acting NIT2 and the positively acting NIT4 regulatory proteins, which might play a role in positioning of chromatin protein. PMID- 8226617 TI - Role of the imp operon of the Streptomyces coelicolor genetic element SLP1: two imp-encoded proteins interact to autoregulate imp expression and control plasmid maintenance. AB - The Streptomyces coelicolor genetic element SLP1 can exist either integrated into the host chromosome or as an autonomously replicating plasmid. The integrated form of SLP1 includes a locus (imp, for inhibition of plasmid maintenance) that can act both in cis and in trans to prevent propagation of SLP1 as an extrachromosomal replicon (S. R. Grant, S. C. Lee, K. Kendall, and S. N. Cohen, Mol. Gen. Genet. 217:324-331, 1989). We report here that a 1.8-kb Eco47III DNA fragment previously shown to encode the Imp+ phenotype contains two genes (impA and impC) that must be expressed in cis to each other and whose products interact functionally and probably physically to interfere with SLP1 plasmid maintenance and repress expression of the imp operon. Partial repression of the imp promoter (P(imp)), which is located immediately 5' of impA, by the 29.7-kDa ImpA protein is enhanced by the impC gene product. Gel shift analysis indicates that ImpA binds to a 16-bp sequence located within the DNA segment containing P(imp) and that ImpC interferes with this binding. Our data suggest that binding of ImpA to the P(imp) region mediates DNA looping in this region. PMID- 8226618 TI - Purification and properties of a nif-specific flavodoxin from the photosynthetic bacterium Rhodobacter capsulatus. AB - A flavodoxin was isolated from iron-sufficient, nitrogen-limited cultures of the photosynthetic bacterium Rhodobacter capsulatus. Its molecular properties, molecular weight, UV-visible absorption spectrum, and amino acid composition suggest that it is similar to the nif-specific flavodoxin, NifF, of Klebsiella pneumoniae. The results of immunoblotting showed that R. capsulatus flavodoxin is nif specific, since it is absent from ammonia-replete cultures and is not synthesized by the mutant strain J61, which lacks a nif-specific regulator (NifR1). Growth of cultures under iron-deficient conditions causes a small amount of flavodoxin to be synthesized under ammonia-replete conditions and increases its synthesis under N2-fixing conditions, suggesting that its synthesis is under a dual system of control with respect to iron and fixed nitrogen availability. Here we show that flavodoxin, when supplemented with catalytic amounts of methyl viologen, is capable of efficiently reducing nitrogenase in an illuminated chloroplast system. Thus, this nif-specific flavodoxin is a potential in vivo electron carrier to nitrogenase; however, its role in the nitrogen fixation process remains to be established. PMID- 8226619 TI - Posttranslational regulation of nitrogenase activity by anaerobiosis and ammonium in Azospirillum brasilense. AB - In the microaerophilic diazotroph Azospirillum brasilense, the addition of fixed nitrogen or a shift to anaerobic conditions leads to a rapid loss of nitrogenase activity due to ADP-ribosylation of dinitrogenase reductase. The product of draT (DRAT) is shown to be necessary for this modification, and the product of draG (DRAG) is shown to be necessary for the removal of the modification upon removal of the stimulus. DRAG and DRAT are themselves subject to posttranslational regulation, and this report identifies features of that regulation. We demonstrate that the activation of DRAT in response to an anaerobic shift is transient but that the duration of DRAT activation in response to added NH4+ varies with the NH4+ concentration. In contrast, DRAG appears to be continuously active under conditions favoring nitrogen fixation. Thus, the activities of DRAG and DRAT are not always coordinately regulated. Finally, our experiments suggest the existence of a temporary period of futile cycling during which DRAT and DRAG are simultaneously adding and removing ADP-ribose from dinitrogenase reductase, immediately following the addition of a negative stimulus. PMID- 8226620 TI - Alanine dehydrogenase (ald) is required for normal sporulation in Bacillus subtilis. AB - The ski22::Tn917lac insertion mutation in Bacillus subtilis was isolated in a screen for mutations that cause a defect in sporulation but are suppressed by the presence or overexpression of the histidine protein kinase encoded by kinA (spoIIJ). The ski22::Tn917lac insertion mutation was in ald, the gene encoding alanine dehydrogenase. Alanine dehydrogenase catalyzes the deamination of alanine to pyruvate and ammonia and is needed for growth when alanine is the sole carbon or nitrogen source. The sporulation defect caused by null mutations in ald was partly relieved by the addition of pyruvate at a high concentration, indicating that the normal role of alanine dehydrogenase in sporulation might be to generate pyruvate to provide an energy source for sporulation. The spoVN::Tn917 mutation was also found to be an allele of ald. Transcription of ald was induced very early during sporulation and by the addition of exogenous alanine during growth. Expression of ald was normal in all of the regulatory mutants tested, including spo0A, spo0K, comA, sigB, and sigD mutants. The only gene in which mutations affected expression of ald was ald itself. This regulation is probably related to the metabolism of alanine. PMID- 8226621 TI - Use of the rep technique for allele replacement to construct mutants with deletions of the pstSCAB-phoU operon: evidence of a new role for the PhoU protein in the phosphate regulon. AB - The phosphate regulon is negatively regulated by the PstSCAB transporter and PhoU protein by a mechanism that may involve protein-protein interaction(s) between them and the Pi sensor protein, PhoR. In order to study such presumed interaction(s), mutants with defined deletions of the pstSCAB-phoU operon were made. This was done by construction of M13 recombinant phage carrying these mutations and by recombination of them onto the chromosome by using a rep host (which cannot replicate M13) for allele replacement. These mutants were used to show that delta (pstSCAB-phoU) and delta (pstB-phoU) mutations abolished Pi uptake by the PstSCAB transporter, as expected, and that delta phoU mutations had no effect on uptake. Unexpectedly, delta phoU mutations had a severe growth defect, and this growth defect was (largely) alleviated by a compensatory mutation in the pstSCAB genes or in the phoBR operon, whose gene products positively regulate expression of the pstSCAB-phoU operon. Because delta phoU mutants that synthesize a functional PstSCAB transporter constitutively grew extremely poorly, the PhoU protein must have a new role, in addition to its role as a negative regulator. A role for the PhoU protein in intracellular Pi metabolism is proposed. Further, our results contradict those of M. Muda, N. N. Rao, and A. Torriani (J. Bacteriol. 174:8057-8064, 1992), who reported that the PhoU protein was required for Pi uptake. PMID- 8226622 TI - Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase. AB - The outer membrane (OM) of Fibrobacter succinogenes was isolated by a combination of salt, sucrose, and water washes from whole cells grown on either glucose or cellulose. The cytoplasmic membrane (CM) was isolated from OM-depleted cells after disruption with a French press. The OM and membrane vesicles isolated from the extracellular culture fluid of cellulose-grown cells had a higher density, much lower succinate dehydrogenase activity, and sodium dodecyl sulfate polyacrylamide gel electrophoresis protein profiles different from those of the CM. The OM from both glucose- and cellulose-grown cells and the extracellular membrane vesicles from cellulose-grown cultures exhibited higher endoglucanase, xylanase, and acetylesterase activities than the CM and other cell fractions. Endoglucanase 2 was absent from the isolated OM fractions of glucose- and cellulose-grown cells and from the extracellular membrane vesicles of cellulose grown cells but was present in the CM and intracellular glycogen granule fractions, while endoglucanase 3 was enriched in the OM. Cellobiosidase was located primarily in the periplasm as previously reported, while cellobiase was mainly present in the glycogen granule fraction of glucose-grown cells and in a nongranular glycogen and CM complex in cellulose-grown cells. The cellobiase was not eluted from glycogen granules by cellobiose, maltose, and maltotriose nor from either the granules or the cell membranes by nondenaturing detergents but was eluted from both glycogen granules and cell membranes by high concentrations of salts. The eluted cellobiase rebound almost quantitatively when diluted and mixed with purified glycogen granules but exhibited a low affinity for Avicel cellulose. Thus, we have documented a method for isolation of OM from F. succinogenes, identified the OM origin of the extracellular membrane vesicles, and located glycanases and cellobiase in membrane and glycogen fractions. PMID- 8226623 TI - Cloning, sequence analysis, and hyperexpression of the genes encoding phosphotransacetylase and acetate kinase from Methanosarcina thermophila. AB - The genes for the acetate-activating enzymes, acetate kinase and phosphotransacetylase (ack and pta), from Methanosarcina thermophila TM-1 were cloned and sequenced. Both genes are present in only one copy per genome, with the pta gene adjacent to and upstream of the ack gene. Consensus archaeal promoter sequences are found upstream of the pta coding region. The pta and ack genes encode predicted polypeptides with molecular masses of 35,198 and 44,482 Da, respectively. A hydropathy plot of the deduced phosphotransacetylase sequence indicates that it is a hydrophobic polypeptides; however, no membrane-spanning domains are evident. Comparison of the amino acid sequences deduced from the M. thermophila and Escherichia coli ack genes indicate similar subunit molecular weights and 44% identity (60% similarity). The comparison also revealed the presence of several conserved arginine, cysteine, and glutamic acid residues. Arginine, cysteine, and glutamic acid residues have previously been implicated at or near the active site of the E. coli acetate kinase. The pta and ack genes were hyperexpressed in E. coli, and the overproduced enzymes were purified to homogeneity with specific activities higher than those of the enzymes previously purified from M. thermophila. The overproduced phosphotransacetylase and acetate kinase migrated at molecular masses of 37,000 and 42,000 Da, respectively. The activity of the acetate kinase is optimal at 65 degrees C and is protected from thermal inactivation by ATP. Diethylpyrocarbonate and phenylglyoxal inhibited acetate kinase activity in a manner consistent with the presence of histidine and arginine residues at or near the active site; however, the thiol-directed reagents 5,5'-dithiobis (2-nitrobenzoic acid) and N-ethylmaleimide were ineffective. PMID- 8226624 TI - The regulatory VirA protein of Agrobacterium tumefaciens does not function at elevated temperatures. AB - Previous studies have shown that Agrobacterium tumefaciens causes tumors on plants only at temperatures below 32 degrees C, and virulence gene expression is specifically inhibited at temperatures above 32 degrees C. We show here that this effect persists even when the virA and virG loci are expressed under the control of a lac promoter whose activity is temperature independent. This finding suggests that one or more steps in the signal transduction process mediated by the VirA and VirG proteins are temperature sensitive. Both the autophosphorylation of VirA and the subsequent transfer of phosphate to VirG are shown to be sensitive to high temperatures (> 32 degrees C), and this correlates with the reduced vir gene expression observed at these temperatures. At temperatures of 32 degrees C and higher, the VirA molecule undergoes a reversible inactivation while the VirG molecule is not affected. vir gene induction is temperature sensitive in an acetosyringone-independent virA mutant background but not in a virG constitutive mutant which is virA and acetosyringone independent. These observations all support the notion that the VirA protein is responsible for the thermosensitivity of vir gene expression. However, an Agrobacterium strain containing a constitutive virG locus still cannot cause tumors on Kalanchoe plants at 32 degrees C. This strain induces normal-size tumors at temperatures up to 30 degrees C, whereas the wild-type Agrobacterium strain produces almost no tumors at 30 degrees C. These results suggest that at temperatures above 32 degrees C, the plant becomes more resistant to infection by A. tumefaciens and/or functions of some other vir gene products are lost in spite of their normal levels of expression. PMID- 8226625 TI - Mycobacteriophage L5 integrase-mediated site-specific integration in vitro. AB - Mycobacteriophage L5, a temperate phage of the mycobacteria, forms stable lysogens in Mycobacterium smegmatis via site-specific integration of the phage genome. Recombination occurs within specific phage and bacterial attachment sites and is catalyzed by the phage-encoded integrase protein in vivo. We describe here the overexpression and purification of L5 integrase and its ability to mediate integrative recombination in vitro. We find that L5 integrase-mediated recombination is greatly stimulated by extracts of M. smegmatis but not by Escherichia coli extracts, purified E. coli integration host factor, or purified HU, indicating the presence of a novel mycobacterial integration host factor. PMID- 8226626 TI - Purification of an SOS repressor from Bacillus subtilis. AB - We have identified in Bacillus subtilis a DNA-binding protein that is functionally analogous to the Escherichia coli LexA protein. We show that the 23 kDa B. subtilis protein binds specifically to the consensus sequence 5' GAACN4GTTC-3' located within the putative promoter regions of four distinct B. subtilis DNA damage-inducible genes: dinA, dinB, dinC, and recA. In RecA+ strains, the protein's specific DNA binding activity was abolished following treatment with mitomycin C; the decrease in DNA binding activity after DNA damage had a half-life of about 5 min and was followed by an increase in SOS gene expression. There was no detectable decrease in DNA binding activity in B. subtilis strains deficient in RecA (recA1, recA4) or otherwise deficient in SOS induction (recM13) following mitomycin C treatment. The addition of purified B. subtilis RecA protein, activated by single-stranded DNA and dATP, abolished the specific DNA binding activity in crude extracts of RecA+ strains and strains deficient in SOS induction. We purified the B. subtilis DNA-binding protein more than 4,000-fold, using an affinity resin in which a 199-bp DNA fragment containing the dinC promoter region was coupled to cellulose. We show that B. subtilis RecA inactivates the DNA binding activity of the purified B. subtilis protein in a reaction that requires single-stranded DNA and nucleoside triphosphate. By analogy with E. coli, our results indicate that the DNA-binding protein is the repressor of the B. subtilis SOS DNA repair system. PMID- 8226627 TI - chpA and chpB, Escherichia coli chromosomal homologs of the pem locus responsible for stable maintenance of plasmid R100. AB - The pem locus is responsible for stable maintenance of plasmid R100 and consists of two genes, pemI and pemK. The pemK gene product is a growth inhibitor, while the pemI gene product is a suppressor of this inhibitory function. We found that the PemI amino acid sequence is homologous to two open reading frames from Escherichia coli called mazE and orf-83, which are located at 60 and 100 min on the chromosome, respectively. We cloned and sequenced these loci and found additional open reading frames, one downstream of each pemI homolog, both of which encode proteins homologous to PemK. The pem locus homolog at 60 min was named chpA and consists of two genes, chpAI and chpAK; the other, at 100 min, was named chpB and consists of two genes, chpBI and chpBK. The distal portion of chpBK was found to be adjacent to the ppa gene that encodes pyrophosphatase, whose map position had not been previously determined. We then demonstrated that the chpAK and chpBK genes encode growth inhibitors, while the chpAI and chpBI genes encode suppressors for the inhibitory function of the ChpAK and ChpBK proteins, respectively. These E. coli pem locus homologs may be involved in regulation of cell growth. PMID- 8226628 TI - Induction by ethanol of alcohol dehydrogenase activity in Acetobacter pasteurianus. AB - The membrane-bound alcohol dehydrogenase (ADH) activity of Acetobacter pasteurianus NCI1380 was enhanced more than 10-fold by the addition of ethanol to the medium. In order to elucidate the mechanism of the ethanol induction, a gene cluster encoding the dehydrogenase and cytochrome c subunits of ADH was cloned from this strain, and its nucleotide sequence was determined. Comparison of the deduced amino acid sequences and the NH2-terminal sequences determined with purified proteins showed that the dehydrogenase and cytochrome c subunits contained typical signal peptides of 35 and 26 amino acids, respectively. Transcriptional analysis of the cloned genes by primer extension revealed that the gene cluster was transcribed from two different promoters upstream from the dehydrogenase gene. One (59 bp upstream of the ATG start codon) of the two promoters was used in the presence of ethanol, whereas the other (232 bp upstream of the ATG start codon) was used in the absence of ethanol. Immunoblot analyses showed that almost the same amounts of the cytochrome c and the 15-kDa subunits were produced in both the presence and absence of ethanol and that the amount of the dehydrogenase subunit localized in the membrane was decreased in the absence of ethanol. This incorrect localization of the dehydrogenase subunit might be one of the factors responsible for the low ADH activity in the absence of ethanol. PMID- 8226629 TI - Oxygen-regulated in vitro transcription of Rhizobium meliloti nifA and fixK genes. AB - Oxygen concentration regulates the expression of nitrogen fixation genes in the symbiotic bacterium Rhizobium meliloti. We demonstrate that two proteins, FixL and FixJ, that belong to the two-component family of regulatory proteins are necessary and sufficient for oxygen-regulated in vitro transcription of the two key regulatory genes, nifA and fixK. We show directly that FixJ is a transcriptional activator, working in conjunction with the RNA polymerase sigma 70 holoenzyme. Addition of FixL122, a soluble form of the sensor FixL protein, to the transcription assay enhanced FixJ transcriptional activity in response to low oxygen concentration. This enhancement of FixJ activity was correlated with FixJ phosphorylation. PMID- 8226630 TI - Genes encoding two isocitrate dehydrogenase isozymes of a psychrophilic bacterium, Vibrio sp. strain ABE-1. AB - The genes coding for two structurally different isocitrate dehydrogenase isozymes (IDH-I and IDH-II) of a psychrophilic bacterium, Vibrio sp. strain ABE-1, were cloned and sequenced. Open reading frames of the genes (icdI and icdII) are 1,248 and 2,229 bp in length, respectively. The amino acid sequences predicted from the open reading frames of icdI and icdII corresponded to the N-terminal amino acid sequences of the purified IDH-I and IDH-II, respectively. No homology was found between the deduced amino acid sequences of the isozymes; however, the IDH-I, a dimeric enzyme, had a high amino acid sequence identity (74.3%) to the Escherichia coli IDH. The deduced amino acid sequence of the IDH-II, a monomeric enzyme, was not related to any known sequence. However, the IDH-II had an amino acid sequence homologous to that of a cyanogen bromide-cleaved peptide containing a putative active-site methionyl residue of the monomeric IDH of Azotobacter vinelandii. The two genes (icdlI and icdII) were found to be tandemly located in the same orientation. Northern (RNA) blot analyses showed that the two genes are transcribed independently. Primer extension experiments located single transcriptional start sites 39 and 96 bp upstream of the start codons of icdI and icdII, respectively. The amount of icdI transcript but not icdII increased when Vibrio sp. strain ABE-1 cells were cultured in acetate minimal medium. PMID- 8226631 TI - Metabolism of dibenzothiophene and naphthalene in Pseudomonas strains: complete DNA sequence of an upper naphthalene catabolic pathway. AB - From a soil isolate, Pseudomonas strain C18, we cloned and sequenced a 9.8-kb DNA fragment that encodes dibenzothiophene-degrading enzymes. Nine open reading frames were identified and designated doxABDEFGHIJ. Collectively, we refer to these genes as the DOX pathway. At the nucleotide level, doxABD are identical to the ndoABC genes that encode naphthalene dioxygenase of Pseudomonas putida. The DoxG protein is 97% identical to NahC (1,2-dihydroxynaphthalene dioxygenase) of P. putida. DoxE has 37% identity with cis-toluene dihydrodiol dehydrogenase. DoxF is similar to the aldehyde dehydrogenases of many organisms. The predicted DoxHIJ proteins have no obvious sequence similarities to known proteins. Gas chromatography with a flame ionization detector and mass spectroscopy confirmed that the DOX proteins convert naphthalene to salicylate and converting phenanthrene to 1-hydroxy-2-naphthoic acid. doxI mutants convert naphthalene to trans-o-hydroxybenzylidenepyruvate, indicating that the DoxI protein is similar to NahE (trans-o-hydroxybenzylidenepyruvate hydratase-aldolase). Comparison of the DOX sequence with restriction maps of cloned naphthalene catabolic pathway (NAH) genes revealed many conserved restriction sites. The DOX gene arrangement is identical to that proposed for NAH, except that the NAH equivalent of doxH has not been recognized. DoxH may be involved in the conversion of 2-hydroxy-4-(2' oxo-3,5-cyclohexadienyl)-buta-2,4-dienoat e to cis-o-hydroxybenzylidenepyruvate. doxJ encodes an enzyme similar to NahD (isomerase). Our findings indicate that a single genetic pathway controls the metabolism of dibenzothiophene, naphthalene, and phenanthrene in strain C18 and that the DOX sequence encodes a complete upper naphthalene catabolic pathway similar to NAH. PMID- 8226632 TI - Early and late responses of TOL promoters to pathway inducers: identification of postexponential promoters in Pseudomonas putida with lacZ-tet bicistronic reporters. AB - Transcriptional lacZ fusions to the Pu and Pm promoters of the TOL (toluene degradation) plasmid inserted in monocopy in the chromosome of Pseudomonas putida showed a very different responsiveness to their respective aromatic effectors regarding growth phase. While a substantial XylS-dependent activation of Pm-lacZ was detected nearly instantly after m-toluate addition, XylR- and xylene-mediated induction of the sigma 54 promoter Pu became significant only after cells slowed down exponential growth and entered stationary phase. When Pu and Pm were fused to lacZ-tet reporters (i.e., promoterless lacZ genes coupled to a tet gene which confers resistance to tetracycline when cotranscribed with the leading gene) instead of lacZ alone, the resulting colonies displayed a distinct phenotype consisting of hyperfluorescence on agar plates after being sprayed with 4 methylumbelliferyl-beta-D-galactoside, simultaneously with being either sensitive (Pu) or resistant (Pm) to tetracycline. To examine whether the same phenotype could be scored in strains carrying transcriptional fusions of the lacZ-tet cassettes to other genes or promoters whose expression is silenced during growth and activated in stationary phase, we constructed mini-Tn5 lacZ-tet transposons for random genetic probing of promoters preferentially active at advanced stages of growth. Chromosomal insertions of this mobile element were selected by means of the constitutive resistance to kanamycin which is also specified by the transposon. A number of kanamycin-resistant colonies which are hyperfluorescent with 4-methylumbelliferyl-beta-D-galactosidase but sensitive to tetracycline and which reached full induction only at postexponential growth stages were obtained. PMID- 8226633 TI - SKN7, a yeast multicopy suppressor of a mutation affecting cell wall beta-glucan assembly, encodes a product with domains homologous to prokaryotic two-component regulators and to heat shock transcription factors. AB - A search for genes which, at elevated copy number, could suppress the growth defect in a strain disrupted at the KRE9 locus has identified the SKN7 gene. SKN7 was mapped to the right arm of chromosome VIII and is predicted to encode a 70 kDa protein, Skn7p, with a region of homology to the DNA binding domain of the Saccharomyces cerevisiae heat shock transcription factor, Hsf1p. Skn7p also has a domain which shows similarity to the prokaryotic receiver modules found on an extensive family of two-component response regulators, including the products of the rcsC and barA genes. SKN7 did not suppress other mutations in the (1-->6) beta-glucan biosynthetic pathway, suggesting that SKN7 does not act as a general bypass suppressor of this glucan. PMID- 8226634 TI - Precursor flux control through targeted chromosomal insertion of the lysine epsilon-aminotransferase (lat) gene in cephamycin C biosynthesis. AB - Targeted gene insertion methodology was used to study the effect of perturbing alpha-aminoadipic acid precursor flux on the overall production rate of beta lactam biosynthesis in Streptomyces clavuligerus. A high-copy-number plasmid containing the lysine epsilon-aminotransferase gene (lat) was constructed and used to transform S. clavuligerus. The resulting recombinant strain (LHM100) contained an additional complete copy of lat located adjacent to the corresponding wild-type gene in the chromosome. Biological activity and production levels of beta-lactam antibiotics were two to five times greater than in wild-type S. clavuligerus. Although levels of lysine epsilon-aminotransferase were elevated fourfold in LHM100, the level of ACV synthetase, whose gene is located just downstream of lat, remained unchanged. These data strongly support the notion that direct perturbation of alpha-aminoadipic acid precursor flux resulted in increased antibiotic production. This strategy represents a successful application of metabolic engineering based on theoretical predictions of precursor flux in a secondary metabolic pathway. PMID- 8226635 TI - Nucleotide sequence and 3'-end deletion studies indicate that the K(+)-uptake protein kup from Escherichia coli is composed of a hydrophobic core linked to a large and partially essential hydrophilic C terminus. AB - The kup (formerly trkD) gene from Escherichia coli encodes a minor K(+)-uptake system. The gene is located just upstream of the rbsDACBK operon at 84.5 min on the chromosome and is transcribed clockwise. kup codes for a 69-kDa protein, which may be composed of two domains. The first 440 amino acid residues appear to form an integral membrane protein that might traverse the cell membrane 12 times. The C-terminal 182 amino acid residues are predicted to form a hydrophilic domain located at the cytoplasmic side of the membrane. Deletion studies from the 3' end of kup showed that removal of almost the complete hydrophilic domain of the protein reduced, but did not abolish, K(+)-uptake activity. PMID- 8226636 TI - Characterization of the Tn5 transposase and inhibitor proteins: a model for the inhibition of transposition. AB - Tn5 is a composite transposon consisting of two IS50 sequences in inverted orientation with respect to a unique, central region encoding several antibiotic resistances. The IS50R element encodes two proteins in the same reading frame which regulate the transposition reaction: the transposase (Tnp), which is required for transposition, and an inhibitor of transposition (Inh). The inhibitor is a naturally occurring deletion variant of Tnp which lacks the N terminal 55 amino acids. In this report, we present the purification of both the Tnp and Inh proteins and an analysis of their DNA binding properties. Purified Tnp, but not Inh, was found to bind specifically to the outside end of Tn5. Inh, however, stimulated the binding activity of Tnp to outside-end DNA and was shown to be present with Tnp in these bound complexes. Inh was also found to exist as a dimer in solution. These results indicate that the N-terminal 55 amino acids of Tnp are required for sequence-specific binding. They also suggest that Inh inhibits transposition by forming mixed oligomers with Tnp which still bind to the ends of the transposon but are defective for later stages of the transposition reaction. PMID- 8226637 TI - Control of gluconeogenic growth by pps and pck in Escherichia coli. AB - It is well-known that Escherichia coli grows more slowly on gluconeogenic carbon sources than on glucose. This phenomenon has been attributed to either energy or monomer limitation. To investigate this problem further, we varied the expression levels of pck, encoding phosphoenolpyruvate carboxykinase (Pck), and pps, encoding phosphoenolpyruvate synthase (Pps). We found that the growth rates of E. coli in minimal medium supplemented with succinate and with pyruvate are limited by the levels of Pck and Pps, respectively. Optimal overexpression of pck or pps increases the unrestricted growth rates on succinate and on pyruvate, respectively, to the same level attained by the wild-type growth rate on glycerol. Since Pps is needed to supply precursors for biosyntheses, we conclude that E. coli growing on pyruvate is limited by monomer supply. However, because pck is required both for biosyntheses and catabolism for cells growing on succinate, it is possible that growth on succinate is limited by both monomer and energy supplies. The growth yield with respect to oxygen remains approximately constant, even though the overproduction of these enzymes enhances gluconeogenic growth. It appears that the constant yield for oxygen is characteristic of efficient growth on a particular substrate and that the yield is already optimal for wild-type strains. Further increases in either Pck or Pps above the optimal levels become growth inhibitory, and the growth yield for oxygen is reduced, indicating less efficient growth. PMID- 8226638 TI - Physical map of the genome of Rhizobium meliloti 1021. AB - A physical map of the genome of Rhizobium meliloti 1021 is presented. The physical sizes of the three replicons in this genome had previously been determined and are as follows: the chromosome, 3.4 Mb; pSym-b, 1.7 Mb; and pSym a, 1.4 Mb. The physical maps for this GC-rich genome contain AT-rich restriction sites for SwaI (5'-TAAATTTA-3'), PacI (5'-TTAATTAA-3'), PmeI (5'-GTTTAAAC-3'), and, for pSym-b, SpeI (5'-ACTAGT-3'). In addition, the endonuclease I-CeuI cleaved the 23S rRNA genes in this genome, and perhaps in most eubacterial genomes. I-CeuI digestion and polymerase chain reaction amplification of rrn regions were used to determine that there are at least three rrn loci in R. meliloti, all of which are located on the chromosome. The orientation of the rrn loci was determined by Southern blotting with probes from rrn sequences located 5' and 3' to the I-CeuI site. The rrn loci are clustered in one part of the chromosome and are oriented so that transcription will occur away from a single point in the circle, as observed for the origin of replication in the Escherichia coli and Salmonella typhimurium chromosomes. Fifteen genes that had been tagged by Tn5 insertion were localized to fragments on the chromosome physical map by using the IS50 as a probe in Southern blots. In addition, glt and gap were placed on the physical map by using Southern hybridization with cloned genes. The fortuitous occurrence of SpecI site in Tn5-233 was used to physically map 10 genetically mapped Tn5-233 integrations on pSym-b and to anchor the physical map to the genetic map. Finally, we demonstrate the usefulness of the map by localizing a total of 12 previously unmapped transposon insertions in the genome. This is the first physical map of the genome of a multireplicon member of the family Rhizobiaceae as well as the first physical map of a Rhizobium chromosome. PMID- 8226639 TI - Cloning, sequencing, and molecular analysis of the sol operon of Clostridium acetobutylicum, a chromosomal locus involved in solventogenesis. AB - A DNA region of Clostridium acetobutylicum contiguous with the adc operon has been cloned and sequenced. Structural genes encoding the acetoacetyl coenzyme A:acetate/butyrate:coenzyme A transferase (ctfB and ctfA) and an alcohol/aldehyde dehydrogenase (adhE) could be identified. These three genes together with a small open reading frame (ORF) of unknown function (upstream of adhE) formed an operon (sol operon), as shown by mRNA analyses. The complete sol operon was transcriptionally induced or derepressed before the onset of solventogenesis, thus confirming earlier results of Northern hybridizations with a ctfB gene probe (U. Gerischer and P. Durre, J. Bacteriol. 174:426-433, 1992). Upstream of the sol operon, we identified two putative promoters that were located in regions with possible stem-loop structures formed by several inverted repeats. The distal promoter P1 showed only minor transcription initiation in solventogenic C. acetobutylicum cells but was recognized in Escherichia coli, presumably because of its high similarity to the sigma 70 consensus sequence. The adhE-proximal promoter P2 directed the major transcription start point in solventogenic C. acetobutylicum but was not recognized in E. coli. The clostridial AdhE showed high similarity to a novel family (type III) of alcohol dehydrogenases. Two other ORFs (ORF 5 and ORF 6) were found on the cloned DNA region that showed no significant similarity to sequences in various available data bases. mRNA studies revealed that ORF 5 formed a monocistronic operon and showed increased expression before onset of solventogenesis. PMID- 8226640 TI - Asymmetric expression of the gyrase B gene from the replication-competent chromosome in the Caulobacter crescentus predivisional cell. AB - The bacterium Caulobacter crescentus undergoes an asymmetric cell division resulting in the formation of two different daughter cells, a motile swarmer cell and a nonmotile stalked cell. These two cell types differ in their program of gene expression, their ability to replicate DNA, and the physical properties of their nucleoids. We show here that two genes, gyrB (encoding the gyrase B subunit) and orf-1, are specifically transcribed from the chromosome in the portion of the predivisional cell destined for the progeny stalked cell. This is in contrast to a subset of flagellar genes which are transcribed from the chromosome in the incipient swarmer portion of the predivisional cell. gyrB and orf-1 are within a newly identified cluster of genes involved in DNA replication and recombination, including dnaN and recF. The transcription of gyrB and orf1 occurs from the replication-competent chromosome in stalked and predivisional cells and is silenced in swarmer cells. We hypothesize that selective silencing of groups of genes in the chromosomes at the swarmer and stalked poles of the predivisional cell results in the different developmental programs and the difference in replicative ability of the two progeny cells. PMID- 8226641 TI - Transcription of ColE1Ap mbeC induced by conjugative plasmids from twelve different incompatibility groups. AB - Although nonconjugative mobilizable plasmids require helping functions of conjugative plasmids in order to be mobilized into recipients, at least some genes from the nonconjugative plasmids may be induced to assist in the DNA transfer process. Conjugative plasmids from 12 different incompatibility groups mobilized the nonconjugative plasmid ColE1Ap between Escherichia coli strains. Introduction of any of the conjugative plasmids into the ColE1Ap-containing strain resulted in an induction of mbeC, the product of which is a component of the mobilization relaxation complex. Each of the conjugative plasmids caused protein to bind specifically to mbe promoter DNA, suggesting a direct regulatory interaction. PMID- 8226642 TI - Regions of maltose-binding protein that influence SecB-dependent and SecA dependent export in Escherichia coli. AB - In Escherichia coli, the efficient export of maltose-binding protein (MBP) is dependent on the chaperone SecB, whereas export of ribose-binding protein (RBP) is SecB independent. To localize the regions of MBP involved in interaction with SecB, hybrids between MBP and RBP in SecB mutant cells were constructed and analyzed. One hybrid consisted of the signal peptide and first third of the mature moiety of MBP, followed by the C-terminal two-thirds of RBP (MBP-RBP112). This hybrid was dependent upon SecB for its efficient export and exhibited a strong export defect in secA mutant cells. A hybrid between RBP and MBP with the same fusion point was also constructed (RBP-MBP116). The RBP-MBP116 hybrid remained SecB independent and only exhibited a partial export defect in secA mutant cells. In addition, MBP species with specific alterations in the early mature region were less dependent on SecB for their efficient export. The export of these altered MBP species was also less affected in secA mutant cells and in cells treated with sodium azide. These results present additional evidence for the targeting role of SecB. PMID- 8226643 TI - The active form of the KorB protein encoded by the Streptomyces plasmid pIJ101 is a processed product that binds differentially to the two promoters it regulates. AB - The korB gene of Streptomyces lividans plasmid pIJ101 is known to encode an autoregulated protein that also represses transcription of a gene, kilB, implicated in pIJ101 transfer and in spreading of the plasmid along mycelia of the recipient. Earlier work has indicated that the primary gene product of korB is a 10-kDa protein predicted from the gene sequence (D.S. Stein and S.N. Cohen, Mol. Gen. Genet. 222:337-344, 1990; S. Zamen H. Richards, and J. Ward, Nuleic Acids Res. 20:3693-3700, 1992). We report here that the 10-kDa KorB protein product is processed in vivo into a 6-kDa peptide that has a 20-fold-greater binding affinity for its operator-promoter target; in addition, the 6-kDa peptide binds differentially to the regulatory regions of the two genes it controls, showing 50-fold-greater affinity for the kilB sequence. While both the processed and unprocessed forms of KorB were observed in Escherichia coli following korB gene expression under control of the bacteriophage T7 promoter, only the 6-kDa peptide was found in S. lividans containing pIJ101, implying that this peptide is normally the biologically active form of KorB. The footprint resulting from KorB binding to the korB operator sequence overlaps the sti locus, which affects pIJ101 copy number and incompatibility as well as the size of zones of inhibited recipient cell growth ("pocks") that form around donor cells during mating. The observed ability of the korB gene product to interact with both sti sequences and the kilB promoter region suggests that it may have a role in coordinating the replication and intramycelial spread of plasmids during and/or following bacterial mating. PMID- 8226644 TI - Mechanism of autophosphorylation of Escherichia coli nitrogen regulator II (NRII or NtrB): trans-phosphorylation between subunits. AB - Nitrogen regulator II (NRII or NtrB) is a homodimeric signal-transducing protein kinase/phosphatase responsible for the transcriptional regulation of the Ntr regulon in Escherichia coli. NRII is a member of a large family of proteins that are part of the related two-component signal transduction systems. We studied the mechanism of NRII autophosphorylation by using purified components. Alteration of the site of NRII autophosphorylation to asparagine (H-139-->N [H139N]) or deletion of the C-terminal 59 amino acids of NRII (ter291) resulted in proteins that were not autophosphorylated upon incubation with ATP. Alteration of glycine 313 to alanine resulted in a protein (G313A) that was phosphorylated to a lesser extent than the wild-type protein. Unlike wild-type NRII and H139N, G313A could not be efficiently cross-linked to [alpha-32P]ATP, suggesting that the G313A mutation affects nucleotide binding. Fusion of maltose-binding protein (MBP) to the N-terminal end of NRII resulted in a protein (MBP-NRII) that autophosphorylated normally. We developed a procedure for forming mixed dimers in vitro from these proteins. In mixed dimers consisting of MBP-NRII and H139N, only the MBP-NRII subunit is phosphorylated. In contrast, in mixed dimers consisting of MBP-NRII and G313A, phosphorylation is predominantly on the G313A subunit. We also demonstrated that the G313A and H139N proteins could complement for the autophosphorylation reaction when they were treated so as to permit the formation of mixed dimers and that the wild-type and H139N proteins could phosphorylate the ter291 protein. These results indicate that the autophosphorylation reaction occurs within the dimer by a trans, intersubunit mechanism in which one subunit binds ATP and phosphorylates the other subunit. PMID- 8226645 TI - Family of glycosyl transferases needed for the synthesis of succinoglycan by Rhizobium meliloti. AB - Rhizobium meliloti produces an acidic exopolysaccharide, termed succinoglycan or EPS I, that is important for invasion of the nodules that it elicits on its host, Medicago sativa. Succinoglycan is a high-molecular-weight polymer composed of repeating octasaccharide subunits. These subunits are synthesized on membrane bound isoprenoid lipid carriers, beginning with a galactose residue followed by seven glucose residues, and modified by the addition of acetate, succinate, and pyruvate. Biochemical characterizations of lipid-linked succinoglycan biosynthetic intermediates from previously identified exo mutant strains have been carried out in our laboratory (T. L. Reuber and G. C. Walker, Cell 74:269 280, 1993) to determine where each mutation blocks the biosynthetic pathway. We have carried out a fine structure genetic analysis of a portion of the cluster of exo genes present on the second symbiotic megaplasmid of R. meliloti and have identified several new genes. In addition, the DNA sequence of 16 kb of the exo cluster was determined and the genetic map was correlated with the DNA sequence. In this paper we present the sequence of a family of glycosyl transferases required for the synthesis of succinoglycan and discuss their functions. PMID- 8226646 TI - Genes needed for the modification, polymerization, export, and processing of succinoglycan by Rhizobium meliloti: a model for succinoglycan biosynthesis. AB - The major acidic exopolysaccharide of Rhizobium meliloti, termed succinoglycan, is required for nodule invasion and possibly nodule development. Succinoglycan is a polymer of octasaccharide subunits composed of one galactose residue, seven glucose residues, and acetyl, succinyl, and pyruvyl modifications, which is synthesized on an isoprenoid lipid carrier. A cluster of exo genes in R. meliloti are required for succinoglycan production, and the biosynthetic roles of their gene products have recently been determined (T.L. Reuber and G. C. Walker, Cell 74:269-280, 1993). Our sequencing of 16 kb of this cluster of exo genes and further genetic analysis of this region resulted in the discovery of several new exo genes and has allowed a correlation of the genetic map with the DNA sequence. In this paper we present the sequences of genes that are required for the addition of the succinyl and pyruvyl modifications to the lipid-linked intermediate and genes required for the polymerization of the octasaccharide subunits or the export of succinoglycan. In addition, on the basis of homologies to known proteins, we suggest that ExoN is a uridine diphosphoglucose pyrophosphorylase and that ExoK is a beta(1,3)-beta (1,4)-glucanase. We propose a model for succinoglycan biosynthesis and processing which assigns roles to the products of nineteen exo genes. PMID- 8226647 TI - purU, a source of formate for purT-dependent phosphoribosyl-N-formylglycinamide synthesis. AB - A gene designated purU has been identified and characterized. purU is adjacent to tyrT at min 27.7 on the Escherichia coli chromosome. The gene codes for a 280 amino-acid protein. The C-terminal segment of PurU from residues 84 to 280 exhibits 27% identity with 5'-phosphoribosylglycinamide (GAR) transformylase, the product of purN. Primer extension mapping and assays of lacZ in a promoter probe vector identified two promoters giving mono- and bi-cistronic purU mRNA. Neither mRNA was regulated by purines. Mutations in either of two pairs of genes are required to block synthesis of 5'-phosphoribosyl-N-formylglycinamide (FGAR) from GAR: purN purT (purT encodes an alternative formate-dependent GAR transformylase) or purN purU. On the basis of the growth of purU, purN, and purU purN mutants, it appears that PurU provides the major source of formate for the purT-dependent synthesis of FGAR. PMID- 8226648 TI - A cytoplasmic protein, NfrC, is required for bacteriophage N4 adsorption. AB - At least four genes are required for irreversible adsorption of bacteriophage N4. nfrA and nfrB have been characterized previously and encode an outer membrane protein and inner membrane protein, respectively. The nfrC gene product is characterized in detail in this study. We have mapped the nfrD locus to min 52 on the Escherichia coli linkage map. Maxicell analysis of nfrC and a null allele (nfrC2) cloned into a high-copy-number plasmid shows its gene product to be 42 kDa in size. We determined the nfrC nucleotide sequence which predicts a gene product of 42 kDa. Western blots (immunoblots) of Escherichia coli proteins after cellular fractionation show NfrC to be a cytoplasmic protein which is required for irreversible bacteriophage N4 adsorption, an event occurring at the cell surface. PMID- 8226649 TI - Two overlapping genes encoding membrane proteins required for bacteriophage N4 adsorption. AB - We present the nucleotide sequences of two genes whose products are required for bacteriophage N4 adsorption. The nfrA gene encodes a 122-kDa outer membrane protein which presumably serves as the phage receptor. The nfrB gene encodes an 85-kDa inner membrane protein and may be a component of the receptor. PMID- 8226650 TI - Bacteriophage P22 transduction of integrated plasmids: single-step cloning of Salmonella typhimurium gene fusions. AB - Transcriptional fusions to Salmonella typhimurium chromosomal genes were constructed by integration of a suicide fusion vector into the chromosome by homologous recombination with random cloned chromosomal fragments. We describe here a transductional method using the generalized transducing phage of S. typhimurium, P22, to clone these fusions directly from the bacterial chromosome, in a single step, without the use of restriction enzymes. In this transduction, the phage packages the chromosomal fragment containing the integrated plasmid. Once introduced into the recipient, the plasmid circularizes by homologous recombination between the duplicated region determined by the cloned fragment. Although RecA mediates the majority of these events, the plasmid can circularize in a recA recipient. However, in this case, the event occurs at a much lower frequency and only when the transduction is done at a high multiplicity of infection. In addition to integrated fusion constructs, we also show that autonomously replicating low-copy-number plasmids can be transduced. In this case, transduction is dependent on homologous recombination between the plasmid and the donor chromosome via cloned sequences, in which the transducing particle effectively traps the integrated plasmid. PMID- 8226651 TI - Isolation and analysis of novel mutants of Escherichia coli prlA (secY). AB - Plasmid libraries of prlA mutants containing single-base-pair changes throughout the gene were generated by in vitro random mutagenesis. The prlA mutations capable of suppressing the secretion defect of LamB caused by mutations in the LamB signal peptide were selected and analyzed. Together with additional mutations generated by site-directed mutagenesis, a number of novel prlA mutations and/or suppressors were identified. These mutations provide the starting points for studying the relationship of structure and function of PrlA in its interaction with LamB and/or other component(s) in the Escherichia coli protein secretion-translocation complex. PMID- 8226652 TI - Antisense transcription of the ftsZ-ftsA gene junction inhibits cell division in Escherichia coli. AB - A 490-bp DNA segment spanning the junction between the ftsA and ftsZ genes inhibits cell division when present in high copy number. We show that this segment contains an antisense promoter and an antisense transcription terminator which define a new gene, stfZ. PMID- 8226653 TI - Antioxidant function of fungal melanin. AB - Polyphenols have been implicated in the virulence and oxidant resistance of Cryptococcus neoformans. Although monomeric polyphenols did not protect against the prooxidant, plumbagin, polymeric dopamine-melanin conferred resistance both to hypochlorite and to permanganate. The physiologic antioxidant capacity conferred by melanin was found to be 21.3 x 10(-15) mole-equivalents per cell, a value which approximates oxidant production by stimulated macrophages. PMID- 8226654 TI - Adaptation of Escherichia coli to the uncoupler of oxidative phosphorylation 2,4 dinitrophenol. AB - Escherichia coli was found to adapt to the uncoupler of oxidative phosphorylation 2,4-dinitrophenol. The rates of synthesis of 53 proteins were increased following exposure to 2,4-dinitrophenol. Adaptation was accelerated when the cofactor pyrroloquinoline quinone was provided in the growth medium. PMID- 8226655 TI - Photolithoautotrophic growth and control of CO2 fixation in Rhodobacter sphaeroides and Rhodospirillum rubrum in the absence of ribulose bisphosphate carboxylase-oxygenase. AB - Rhodospirillum rubrum and Rhodobacter sphaeroides were shown to be capable of photolithoautotrophic growth in the absence of the reductive pentose phosphate (Calvin) cycle. Ribulose 1,5-bisphosphate carboxylase-oxygenase (RubisCO) deletion strains were incapable of photolithoautotrophic growth using hydrogen as an electron donor but were able to grow in the absence of organic carbon using less reduced inorganic electron donors, i.e., thiosulfate or sulfide. Wild-type R. rubrum grown in the presence of thiosulfate contained RubisCO levels that were 50-fold lower compared with those in cells growth with hydrogen as an electron donor without substantially influencing rates of photolithoautotrophic growth. These results suggest there are two independent CO2 fixation pathways that support photolithoautotrophic growth in purple nonsulfur photosynthetic bacteria, indicating that these organisms have developed sophisticated control mechanisms to regulate the flow of carbon from CO2 through these separate pathways. PMID- 8226656 TI - Use of Salmonella phage P22 for transduction in Escherichia coli. AB - A cosmid (pPR1347) carrying both the rfb gene cluster and the rfc gene of a Salmonella group B serovar has been constructed; Escherichia coli K-12 strains carrying this cosmid produce long-chain O antigen, are sensitive to phage P22, and can be transduced by P22. Some of the benefits of P22 transduction are now available for studying E. coli and potentially other genera. PMID- 8226657 TI - The hydrophobic repeated domain of the Clostridium cellulovorans cellulose binding protein (CbpA) has specific interactions with endoglucanases. AB - We overexpressed one of the hydrophobic repeated domains (HBDs) (110 amino acid residues) of the cellulose-binding protein (CbpA) from Clostridium cellulovorans by making a hybrid protein with the Escherichia coli maltose-binding protein (MalE). The HBD was purified to homogeneity, and interactions between the HBD and endoglucanases were analyzed by a novel interaction Western blotting (immunoblotting) method. The HBD had specific interactions with endoglucanases (EngB and EngD) from C. cellulovorans. These results indicated that the HBD was an endoglucanase binding site of CbpA. PMID- 8226658 TI - Polarized cells, polar actions. AB - The recognition of polar bacterial organization is just emerging. The examples of polar localization given here are from a variety of bacterial species and concern a disparate array of cellular functions. A number of well-characterized instances of polar localization of bacterial proteins, including the chemoreceptor complex in both C. crescentus and E. coli, the maltose-binding protein in E. coli, the B. japonicum surface attachment proteins, and the actin tail of L. monocytogenes within a mammalian cell, involve proteins or protein complexes that facilitate bacterial interaction with the environment, either the extracellular milieux or that within a plant or mammalian host. The significance of this observation remains unclear. Polarity in bacteria poses many problems, including the necessity for a mechanism for asymmetrically distributing proteins as well as a mechanism by which polar localization is maintained. Large structures, such as a flagellum, are anchored at the pole by means of the basal body that traverses the peptidoglycan wall. But for proteins and small complexes, whether in the periplasm or the membrane, one must invoke a mechanism that prevents the diffusion of these proteins away from the cell pole. Perhaps the periplasmic proteins are retained at the pole by the presence of the periseptal annulus (35). The constraining features for membrane components are not known. For large aggregates, such as the clusters of MCP, CheA, and CheW complexes, perhaps the size of the aggregate alone prevents displacement. In most cases of cellular asymmetry, bacteria are able to discriminate between the new pole and the old pole and to utilize this information for localization specificity. The maturation of new pole to old pole appears to be a common theme as well. Given numerous examples reported thus far, we propose that bacterial polarity displays specific rules and is a more general phenomenon than has been previously recognized. PMID- 8226659 TI - Characterization of the gene encoding an intracellular proteinase inhibitor of Bacillus subtilis and its role in regulation of the major intracellular proteinase. AB - The gene (ipi) for an intracellular proteinase inhibitor (BsuPI) from Bacillus subtilis was cloned and found to encode a polypeptide consisting of 119 amino acids with no cysteine residues. The deduced amino acid sequence contained the N terminal amino acid sequence of the inhibitor, which was chemically determined previously, and showed no significant homology to any other proteinase inhibitors. Analysis of the transcription initiation site and mRNA showed that the ipi gene formed an operon with an upstream open reading frame with an unknown function. The transcriptional control of ipi gene expression was demonstrated by Northern (RNA) blot analysis, and the time course of transcriptional enhancement roughly corresponded to the results observed at the protein level. Strains in which the ipi gene was disrupted or in which BsuPI was overexpressed constitutively sporulated normally. Analysis of the time course of production of the intracellular proteinase and proteinase inhibitor in these strains suggested that BsuPI directly regulated the major intracellular proteinase (ISP-1) activity in vivo. PMID- 8226660 TI - Conditional synthesis and utilization of 1,5-anhydroglucitol in Escherichia coli. AB - A cyclic polyol, 1,5-anhydro-D-glucitol (AG), is widely detected in most organisms, although little is known about its metabolism and physiological roles. The present study demonstrates the synthesis of AG in Escherichia coli C600. The major portion of the synthesized AG was indicated to be derived from glucose retaining all the six carbon atoms, and only 5% was attributed to AG synthesized from C3 compounds. AG synthesis is apparent in an early stage of the stationary phase, and accumulation is transient both in cells and in medium. Evidence is also presented for AG uptake and metabolism and for effects of cyclic AMP. PMID- 8226661 TI - The Haemophilus influenzae adenylate cyclase gene: cloning, sequence, and essential role in competence. AB - Competence for transformation in Haemophilus influenzae is stimulated by cyclic AMP (cAMP) and requires the cAMP-dependent catabolite regulatory protein CRP. Thus, understanding the control of competence will require understanding how cAMP levels are regulated. As a first step, we have cloned the H. influenzae adenylate cyclase gene (cya) by complementing the Lac- phenotype of delta cya Escherichia coli. Its sequence specifies an 843-amino-acid protein which has significant identity to other known bacterial adenylate cyclases (41 to 43% and 61% identical to the cya genes of enteric bacteria and of Pasteurella multocida, respectively). As seen in other bacterial cya genes, there is evidence for regulation similar to that demonstrated for E. coli: the presence of a strong consensus CRP binding site within the promoter of the gene may provide feedback control of cAMP levels by repressing cya transcription, and translation may be limited by the weak ribosome binding site and by initiation of protein synthesis with GUG rather than AUG or the UUG used in other bacterial cya genes. We confirmed the essential role of cAMP in competence by constructing and characterizing H. influenzae cya mutants. This strain failed to develop competence either spontaneously or after transfer to a competence-inducing medium. However, it became as competent as its wild-type parent in the presence of exogenous cAMP. This result suggests that the failure of exogenously added cAMP to induce optimum competence in wild-type cells is not due to a limitation to the entry of cAMP into the cells. Rather, it strongly favors models in which competence induction requires both an increase in intracellular cAMP and a second as yet unidentified regulatory event. H. influenzae strains mutant in cya or crp were unable to ferment xylose or ribose. This confirms that influenzae, like E. coli, uses cAMP and CRP to regulate nutrient uptake and utilization and lends increasing support to the hypothesis that DNA uptake is mechanism of nutrient acquisition. PMID- 8226662 TI - Cloning and characterization of the RNA polymerase alpha-subunit operon of Chlamydia trachomatis. AB - We have cloned the chlamydial operon that encodes the initiation factor IF1, the ribosomal proteins L36, S13, and S11, and the alpha subunit of RNA polymerase. The genes for S11 and alpha are closely linked in Escherichia coli, Bacillus subtilis, and plant chloroplast genomes, and this arrangement is conserved in Chlamydia spp. The S11 ribosomal protein gene potentially encodes a protein of 125 amino acids with 41 to 42% identity over its entire length to its E. coli and B. subtilis homologs; the gene encoding the alpha subunit specifies a protein of 322 amino acids with 25 to 30% identity over its entire length to its E. coli and B. subtilis homologs. In a T7-based expression system in E. coli, the chlamydial alpha gene directed the synthesis of a 36-kDa protein. Mapping of the chlamydial mRNA transcript by RNase protection studies and by a combination of reverse transcription and the polymerase chain reaction demonstrates that IF1, L36, S13, S11, and alpha are transcribed as a polycistronic transcript. PMID- 8226663 TI - Stress response of Escherichia coli to elevated hydrostatic pressure. AB - The response of exponentially growing cultures of Escherichia coli to abrupt shifts in hydrostatic pressure was studied. A pressure upshift to 546 atm (55,304 kPa) of hydrostatic pressure profoundly perturbed cell division, nucleoid structure, and the total rate of protein synthesis. The number of polypeptides synthesized at increased pressure was greatly reduced, and many proteins exhibited elevated rates of synthesis relative to total protein synthesis. We designated the latter proteins pressure-induced proteins (PIPs). The PIP response was transient, with the largest induction occurring approximately 60 to 90 min postshift. Fifty-five PIPs were identified. Many of these proteins are also induced by heat shock or cold shock. The PIP demonstrating the greatest pressure induction was a basic protein of 15.6 kDa. High pressure inhibits growth but does not inhibit the synthesis of stringently controlled proteins. Cold shock is the only additional signal which has been found to elicit this type of response. These data indicate that elevated pressure induces a unique stress response in E. coli, the further characterization of which could be useful in delineating its inhibitory nature. PMID- 8226664 TI - Cotranscription of two genes necessary for ribosomal protein L11 methylation (prmA) and pantothenate transport (panF) in Escherichia coli K-12. AB - Genetic complementation and enzyme assays have shown that the DNA region between panF, which encodes pantothenate permease, and orf1, the first gene of the fis operon, encodes prmA, the genetic determinant for the ribosomal protein L11 methyltransferase. Sequencing of this region identified one long open reading frame that encodes a protein of 31,830 Da and corresponds to the prmA gene. We found, both in vivo and in vitro, that prmA is expressed from promoters located upstream of panF and thus that the panF and prmA genes constitute a bifunctional operon. We located the major 3' end of prmA transcripts 90 nucleotides downstream of the stop codon of prmA in the DNA region upstream of the fis operon, a region implicated in the control of the expression of the fis operon. Although no promoter activity was detected immediately upstream of prmA, S1 mapping detected 5' ends of mRNA in this region, implying that some mRNA processing occurs within the bicistronic panF-prmA mRNA. PMID- 8226665 TI - Variations in the number of ribosomal DNA units in morphological mutants and normal strains of Candida albicans and in normal strains of Saccharomyces cerevisiae. AB - Naturally occurring strains of Candida albicans are opportunistic pathogens that lack a sexual cycle and that are usually diploids with eight pairs of chromosomes. C. albicans spontaneously gives rise to a high frequency of colonial morphology mutants with altered electrophoretic karyotypes, involving one or more of their chromosomes. However, the most frequent changes involve chromosome VIII, which contains the genes coding for ribosomal DNA (rDNA) units. We have used restriction fragment lengths to analyze the number and physical array of the rDNA units on chromosome VIII in four normal clinical strains and seven morphological mutants derived spontaneously from one of the clinical isolates. HindIII does not cleave the rDNA repeats and liberates the tandem rDNA cluster from each homolog of chromosome VIII as a single fragment, whereas the cleavage at a single site by NotI reveals the size of the single rDNA unit. All clinical strains and morphological mutants differed greatly in the number of rDNA units per cluster and per cell. The four clinical isolates differed additionally among themselves by the size of the single rDNA unit. For a total of 25 chromosome VIII homologs in a total of 11 strains considered, the variability of chromosome VIII was exclusively due to the length of rDNA clusters (or the number of rDNA units) in approximately 92% of the cases, whereas the others involved other rearrangements of chromosome VIII. Only slight variations in the number of rDNA units were observed among 10 random C. albicans subclones and 10 random Saccharomyces cerevisiae subclones grown for a prolonged time at 22 degrees C. However, when grown faster at optimal temperatures of 37 and 30 degrees C, respectively, both fungi accumulated higher numbers of rDNA units, suggesting that this condition is selected for in rapidly growing cells. The morphological mutants, in comparison with the C. albicans subclones, contained a markedly wider distribution of the number of rDNA units, suggesting that a distinct process may be involved in altering the number of rDNA units in these mutants. PMID- 8226666 TI - Two global regulatory systems (Crp and Arc) control the cobalamin/propanediol regulon of Salmonella typhimurium. AB - The genes for cobalamin (vitamin B12) biosynthesis (cob) are coregulated with genes for degradation of propanediol (pdu). Both the cob and pdu operons are induced by propanediol by means of a positive regulatory protein, PocR. This coregulation of a synthetic and a degradative pathway reflects the fact that vitamin B12 is a required cofactor for the first enzyme in propanediol breakdown. The cob/pdu regulon is induced by propanediol under two sets of growth conditions, i.e., during aerobic respiration of a poor carbon source and during anaerobic growth. We provide evidence that, under aerobic conditions, the Crp/cyclic AMP system is needed for all induction of the pocR, cob, and pdu genes. Anaerobically, the Crp/cyclic AMP and ArcA/ArcB systems act additively to support induction of the same three transcription units. The fact that these global control systems affect expression of the gene for the positive regulatory protein (pocR) as well as the pdu and cob operons is consistent with our previous suggestion that these two global controls may act directly only on the pocR gene; their control over the cob and pdu operons may be an indirect consequence of their effect on the level of PocR activator protein. The reported experiments were made possible by the observation that pyruvate supports aerobic growth of all of the mutants tested (cya, crp, arcA, and arcB); pyruvate also supports anaerobic growth of these mutants if the alternative electron acceptor, fumarate, is provided. By using pyruvate as a carbon source, it was possible to grow all of these mutant strains under identical conditions and compare their expression of the cob/pdu regulon. The role of Crp in control of vitamin B12 synthesis suggests that the major role of vitamin B12 in Salmonella spp. is in catabolism of carbon sources; the coregulation of the cob and pdu operons suggests that propanediol is the major vitamin B12-dependent carbon source. PMID- 8226667 TI - tRNA modification activity is necessary for Tet(M)-mediated tetracycline resistance. AB - Tet(M) protein interacts with the protein biosynthetic machinery to render this process resistant to the tetracycline in vivo and in vitro (V. Burdett, J. Biol. Chem. 266:2872-2877, 1991). To understand this process more completely, a mutant of Escherichia coli which is altered in the ability of Tet(M) to confer resistance has been identified. This mutation maps to miaA and displays phenotypes characteristic of previously isolated miaA mutations. The miaA gene product modifies A37 adjacent to the anticodon of several tRNA species. Both the mutant isolated in this work and previously isolated miaA mutants confer tetracycline sensitivity in the presence of functional Tet(M), both share a slow growth phenotype, and in neither case is a wild-type phenotype restored in trans by F'112 carrying the 89- to 98-min region of the chromosome. These similar phenotypes further substantiate the assignment of the mutation described here to the miaA locus. PMID- 8226668 TI - Kinetics of appearance and disappearance of classes of bacterial ice nuclei support an aggregation model for ice nucleus assembly. AB - The kinetics of appearance and disappearance of three classes of ice nuclei in Pseudomonas syringae was investigated under conditions where high-level expression of the ice nucleation phenotype was obtained. The appearance of types 1, 2, and 3 ice nuclei, catalyzing nucleation at -2 to -5, -5 to -7, and -7 to 10 degrees C, respectively, was investigated during low-temperature induction in wild-type strains and in a unique, detergent-sensitive mutant that contained no type 3 ice nuclei when grown at 32 degrees C. Nuclei appeared in the following order: type 3, then type 2 and type 1. The disappearance of classes of ice nuclei was monitored during high-temperature treatment of fully induced cells; nuclei disappeared in the order type 1, type 2, and type 3. Although analysis of nucleation events is complicated by masking and unmasking of ice sites in the same cells, these temporal sequences of ice nucleus appearance or disappearance are consistent with an aggregation model for ice nucleus assembly (A. G. Govindarajan and S. E. Lindow, Proc. Natl. Acad. Sci. USA 85:1334-1338, 1988; G. Warren and P. Wolber, Mol. Microbiol. 5:239-243, 1991). PMID- 8226669 TI - Bradyrhizobium japonicum delta-aminolevulinic acid dehydratase is essential for symbiosis with soybean and contains a novel metal-binding domain. AB - The Bradyrhizobium japonicum hemA gene product delta-aminolevulinic acid (ALA) synthase is not required for symbiosis of that bacterium with soybean. Hence, the essentiality of the subsequent heme synthesis enzyme, ALA dehydratase, was examined. The B. japonicum ALA dehydratase gene, termed hemB, was isolated and identified on the basis of its ability to confer hemin prototrophy and enzyme activity on an Escherichia coli hemB mutant, and it encoded a protein that was highly homologous to ALA dehydratases from diverse organisms. A novel metal binding domain in the B. japonicum ALA dehydratase was identified that is a structural composite of the Mg(2+)-binding domain found in plant ALA dehydratases and the Zn(2+)-binding region of nonplant ALA dehydratases. Enzyme activity in dialyzed extracts of cells that overexpressed the hemB gene was reconstituted by the addition of Mg2+ but not by addition of Zn2+, indicating that the B. japonicum ALA dehydratase is similar to the plant enzymes with respect to its metal requirement. Unlike the B. japonicum hemA mutant, the hemB mutant strain KP32 elicited undeveloped nodules on soybean, indicated by the lack of nitrogen fixation activity and plant hemoglobin. We conclude that the hemB gene is required for nodule development and propose that B. japonicum ALA dehydratase is the first essential bacterial enzyme for B. japonicum heme synthesis in soybean root nodules. In addition, we postulate that ALA is the only heme intermediate that can be translocated from the plant to the endosymbiont to support bacterial heme synthesis in nodules. PMID- 8226670 TI - Anaerobic control of denitrification in Pseudomonas stutzeri escapes mutagenesis of an fnr-like gene. AB - The synthesis of proteins necessary for the respiratory reduction of nitrate to dinitrogen is induced in most denitrifying bacteria by a shift to anaerobiosis. A homolog of the fur gene, which encodes a redox-active transcriptional activator in Escherichia coli, was isolated from Pseudomonas stutzeri by using the anr gene of Pseudomonas aeruginosa as the hybridization probe (R. G. Sawers, Mol. Microbiol. 5:1469-1481, 1991). The coding region was located on a 3-kb SmaI fragment. An open reading frame of 735 nucleotides, designated fnrA, had the coding potential for a protein of 244 amino acids (M(r) = 27,089) with 51.2% positional identity to the Fnr protein of E. coli and 86.1% to the Anr protein of P. aeruginosa. The fnrA gene gave a single transcript of 0.85 kb and complemented nitrate-dependent anaerobic growth of an fnr deletion mutant of E. coli. An open reading frame immediately downstream of fnrA encoded adenine phosphoribosyltransferase (EC 2.4.2.7). Mutations in fnrA were generated in vitro by insertional mutagenesis followed by gene replacement. Gene inactivation was shown by loss of the fnrA transcript and detection of an arginine deiminase (EC 3.5.3.6)-negative phenotype in the mutants. However, neither the enzymatic activities nor the levels of anaerobic expression of the respiratory enzymes nitrate reductase (EC 1.7.99.4), nitrate reductase (EC 1.9.3.2), NO reductase (EC 1.7.99.7), and N2O reductase (EC 1.7.99.6) were changed in fnrA mutants versus the P. stutzeri wild type. A promoter-probe vector for Fnr-dependent transcription was activated anaerobically in the fnrA mutants, suggesting the existence of a second Fnr homolog in the same bacterium. The Fnr-binding motifs, apparent in the promoter region of genes encoding denitrification components of P. stutzeri, are likely to be recognized by this second Fnr homolog. Preliminary evidence indicates also the presence of the catabolite activator protein, Crp, in P. stutzeri. PMID- 8226671 TI - DNA polymerase I and the bypassing of RecA dependence of constitutive stable DNA replication in Escherichia coli rnhA mutants. AB - In Escherichia coli rnhA mutants, several normally repressed origins (oriK sites) of DNA replication are activated. The type of DNA replication initiated from these origins, termed constitutive stable DNA replication, does not require DnaA protein or the oriC site, which are essential for normal DNA replication. It requires active RecA protein. We previously found that the lexA71(Def)::Tn5 mutation can suppress this RecA requirement and postulated that the derepression of a LexA regulon gene(s) leads to the activation of a bypass pathway, Rip (for RecA-independent process). In this study, we isolated a miniTn10spc insertion mutant that abolishes the ability of the lexA(Def) mutation to suppress the RecA requirement of constitutive stable DNA replication. Cloning and DNA sequencing analysis of the mutant revealed that the insertion occurs at the 3' end of the coding region of the polA gene, which encodes DNA polymerase I. The mutant allele, designated polA25::miniTn10spc, is expected to abolish the polymerization activity but not the 5'-->3' or 3'-->5' exonuclease activity. Thus, the Rip bypass pathway requires active DNA polymerase I. Since the lethal combination of recA(Def) and polA25::miniTn10spc could be suppressed by derepression of the LexA regulon only when DNA replication is driven by the oriC system, it was suggested that the bypass pathway has a specific requirement for DNA polymerase I at the initiation step in the absence of RecA. An accompanying paper (Y. Cao and T. Kogoma, J. Bacteriol. 175:7254-7259, 1993) describes experiments to determine which activities of DNA polymerase I are required at the initiation step and discusses possible roles for DNA polymerase in the Rip bypass pathway. PMID- 8226672 TI - Requirement for the polymerization and 5'-->3' exonuclease activities of DNA polymerase I in initiation of DNA replication at oriK sites in the absence of RecA in Escherichia coli rnhA mutants. AB - In previous studies, we found that the requirement for RecA protein in constitutive stable DNA replication (cSDR) can be bypassed by derepression of the LexA regulon and that DNA polymerase I (DNA PolI) is essential for this Rip (RecA independent process) pathway of cSDR (Y. Cao, R. R. Rowland, and T. Kogoma, J. Bacteriol. 175:7247-7253, 1993). In this study, the role of DNA PolI in the Rip pathway was further examined. By using F' plasmids carrying different parts of the polA gene, a series of complementation tests was carried out to investigate the requirement for the three enzymatic activities, polymerization, 3'-->5' exonuclease, and 5'-->3' exonuclease activities, of DNA PolI. The result indicated that both the 5'-->3' exonuclease and polymerization activities of DNA PolI are essential for bypassing the requirement for RecA in cSDR but that the 3' ->5' exonuclease activity can be dispensed with. Complementation experiments with rat DNA Pol beta also supported the hypothesis that a nick translation activity is probably involved in cSDR in the absence of RecA. An analysis of DNA synthesis suggested that DNA PolI is involved in the initiation but not the elongation stage of cSDR. Moreover, the dnaE293(Ts) mutation was shown to render the bypass replication temperature sensitive despite the presence of active DNA PolI, suggesting that DNA PolIII is responsible for the elongation stage of the Rip pathway. A model which describes the possible roles of RecA in cSDR and the possible function of DNA PolI in the Rip pathway is proposed. PMID- 8226673 TI - Comparative analysis of C3 and botulinal neurotoxin genes and their environment in Clostridium botulinum types C and D. AB - The C3 exoenzyme gene is located on a bacteriophage in Clostridium botulinum types C and D (M. R. Popoff, D. Hauser, P. Boquet, M. W. Eklund, and D. M. Gill, Infect. Immun. 59:3673-3679, 1991). A derivative CN phage from phage C of C. botulinum Stockholm (C-St) (K. Oguma, H. Iida, and K. Inoue, Jpn. J. Microbiol. 19:167-172, 1975), isolated as neurotoxin negative, also does not produce exoenzyme C3. The botulinal neurotoxin C1 gene is present on the CN phage but contains a stop mutation in the DNA region encoding the N-terminal part of the heavy chain (codon 553). The putative truncated botulinal neurotoxin C1 protein was not recovered in a C. botulinum strain harboring the CN phage. We found that the C3 gene is localized on a 21.5-kbp DNA fragment flanked by the core motif 5' AAGGAG-3' in DNAs of phage C of C. botulinum 468 (C-468), C-St phage, and phage D of C. botulinum 1873 (D-1873). The 21.5-kbp DNA fragment is deleted in CN phage DNA, and the motif 5'-AAGGAG-3' is present only in one copy at the deletion junction, but the deletion in the CN phage could be nonspecific, since this phage was obtained by nitrosoguanidine treatment. These findings could indicate that the C3 gene is localized on a 21.5-kbp mobile element. C. botulinum type C strain 003-9 produces a C3 exoenzyme (Y. Nemoto, T. Namba, S. Kozaki, and S. Narumiya, J. Biol. Chem. 266:19312-19319, 1991), and Staphylococcus aureus E1 produces a related C3 enzyme which is named epidernmal cell differentiation inhibitor (S. Inoue, M. Sugai, Y. Murooka, S. Y. Paik, Y. M. Hong, H. Oghai, and H. Suginaka, Biochem. Biophys. Res. Comm. 174:459-464, 1991) and which shares 80.6 and 56.6% similarity, respectively with the C3 enzymes from C-468 or C-St and D-1873 phages athe amino acid level. The features of the putative 21.5-kbp transposon were not found in C. botulinum 003-9 and S. aureus E1, as determined by analysis of the C3 and epidermal cell differentiation inhibitor gene-flanking DNA regions. These data suggest a common ancestral origin and divergent evolution of the C3 genes in these three groups of bacterial strains and dissemination of a 21.5-kbp element carrying the C3 gene C-468, C-St, and D-1873 phages. PMID- 8226674 TI - Structural organization, nucleotide sequence, and regulation of the Haemophilus influenzae rec-1+ gene. AB - The Haemophilus influenzae rec-1+ protein plays a central role in DNA metabolism, participating in general homologous recombination, recombinational (postreplication) DNA repair, and prophage induction. Although many H. influenzae rec-1 mutants have been phenotypically characterized, little is known about the rec-1+ gene at the molecular level. In this study, we present the genetic organization of the rec-1+ locus, the DNA sequence of rec-1+, and studies of the transcriptional regulation of rec-1+ during cellular assault by DNA-damaging agents and during the induction of competence for genetic transformation. Although little is known about promoter structure in H. influenzae, we identified a potential rec-1+ promoter that is identical in 11 of 12 positions to the bacterial sigma 70-dependent promoter consensus sequence. Results from a primer extension analysis revealed that the start site of rec-1+ transcription is centered 6 nucleotides downstream of this promoter. We identified potential DNA binding sites in the rec-1+ gene for LexA, integration host factor, and cyclic AMP receptor protein. We obtained evidence that at least one of the proposed cyclic AMP receptor protein binding sites is active in modulating rec-1+ transcription. This finding makes rec-1+ control circuitry novel among recA+ homologs. Two H. influenzae DNA uptake sequences that may function as a transcription termination signal were identified in inverted orientations at the end of the rec-1+ coding sequence. In addition, we report the first use of the Escherichia coli lacZ operon fusion technique in H. influenzae to study the transcriptional control of rec-1+. Our results indicate that rec-1+ is transcriptionally induced about threefold during DNA-damaging events. Furthermore, we show that rec-1+ can substitute for recA+ in E. coli to modulate SOS induction of dinB1 expression. Surprisingly, although 5% of the H. influenzae genome is in the form of single-stranded DNA during competence for genetic transformation, an event that could be a potent SOS-inducing signal, we failed to detect significant changes in rec-1+ transcription during the induction of genetic competence. PMID- 8226675 TI - The uraA locus and homologous recombination in Mycobacterium bovis BCG. AB - Molecular genetic manipulation of mycobacteria would benefit from the isolation of mycobacterial genes that could serve both as genetic markers and as sequences used to target homologous integration of recombinant DNA into the genome. We isolated the Mycobacterium bovis BCG gene encoding orotidine-5'-monophosphate decarboxylase (OMP-DCase) by complementing an Escherichia coli mutant defective in this activity. The BCG OMP-DCase gene (uraA) and the flanking DNA were sequenced. The predicted BCG OMP-DCase protein sequence is closely related to the Myxococcus xanthus OMP-DCase and more distantly related to the other known prokaryotic and eukaryotic OMP-DCases. To investigate whether homologous integration can occur in M. bovis BCG, an improved protocol for transformation of BCG was developed and a linear fragment of mycobacterial DNA containing the uraA locus, marked with a kanamycin resistance gene, was introduced into BCG cells by electroporation. The kanamycin-resistant BCG transformants all contained vector DNA integrated into the genome. The marked DNA had integrated into the homologous uraA locus in approximately 20% of the transformants. These results have implications for understanding the role of mycobacterial genes in disease pathogenesis and for the genetic engineering of improved mycobacterial vaccines. PMID- 8226676 TI - dcp gene of Escherichia coli: cloning, sequencing, transcript mapping, and characterization of the gene product. AB - Dipeptidyl carboxypeptidase is a C-terminal exopeptidase of Escherichia coli. We have isolated the respective gene, dcp, from a low-copy-number plasmid library by its ability to complement a dcp mutation preventing the utilization of the unique substrate N-benzoyl-L-glycyl-L-histidyl-L-leucine. Sequence analysis of a 2.9-kb DNA fragment revealed an open reading frame of 2,043 nucleotides which was assigned to the dcp gene by N-terminal amino acid sequencing and electrophoretic molecular mass determination of the purified dcp product. Transcript mapping by primer extension and S1 protection experiments verified the physiological significance of potential initiation and termination signals for dcp transcription and allowed the identification of a single species of monocistronic dcp mRNA. The codon usage pattern and the effects of elevated gene copy number indicated a relatively low level of dcp expression. The predicted amino acid sequence of dipeptidyl carboxypeptidase, containing a potential zinc-binding site, is highly homologous (78.8%) to the corresponding enzyme from Salmonella typhimurium. It also displays significant homology to the products of the S. typhimurium opdA and the E. coli prlC genes and to some metalloproteases from rats and Saccharomyces cerevisiae. No potential export signals could be inferred from the amino acid sequence. Dipeptidyl carboxypeptidase was enriched 80-fold from crude extracts of E. coli and used to investigate some of its biochemical and biophysical properties. PMID- 8226677 TI - The Vibrio fischeri luminescence gene activator LuxR is a membrane-associated protein. AB - The Vibrio fischeri luminescence (lux) genes are activated at sufficiently high culture densities by the transcriptional activator LuxR in combination with a diffusible signal compound termed autoinducer. We have used antibodies directed against LuxR in immunoprecipitation experiments to study the subcellular location of this transcription factor. The LuxR polypeptide was detected in membranes and not in the soluble pool of cytoplasmic proteins from V. fischeri. LuxR was not released from the membranes by 0.6 M KCl or by the nonionic detergents Nonidet P 40, N-octyl-beta-D-glucopyranoside, and Triton X-100. LuxR and a number of other V. fischeri proteins were released from the membranes by EDTA. The autoinducer had no detectable influence on the subcellular location of LuxR. In spheroplasts, neither the abundance nor the molecular mass of the LuxR antigen was influenced by treatment with proteinase K. Together with other information, these results indicate that LuxR is an amphipathic protein that is associated with the cytoplasmic membrane of V. fischeri. PMID- 8226678 TI - Characterization of 2,2',3-trihydroxybiphenyl dioxygenase, an extradiol dioxygenase from the dibenzofuran- and dibenzo-p-dioxin-degrading bacterium Sphingomonas sp. strain RW1. AB - A key enzyme in the degradation pathways of dibenzo-p-dioxin and dibenzofuran, namely, 2,2',3-trihydroxybiphenyl dioxygenase, which is responsible for meta cleavage of the first aromatic ring, has been genetically and biochemically analyzed. The dbfB gene of this enzyme has been cloned from a cosmid library of the dibenzo-p-dioxin- and dibenzofuran-degrading bacterium Sphingomonas sp. strain RW1 (R. M. Wittich, H. Wilkes, V. Sinnwell, W. Francke, and P. Fortnagel, Appl. Environ. Microbiol. 58:1005-1010, 1992) and sequenced. The amino acid sequence of this enzyme is typical of those of extradiol dioxygenases. This enzyme, which is extremely oxygen labile, was purified anaerobically to apparent homogeneity from an Escherichia coli strain that had been engineered to hyperexpress dbfB. Unlike most extradiol dioxygenases, which have an oligomeric quaternary structure, the 2,2',3-trihydroxybiphenyl dioxygenase is a monomeric protein. Kinetic measurements with the purified enzyme produced similar Km values for 2,2',3-trihydroxybiphenyl and 2,3-dihydroxybiphenyl, and both of these compounds exhibited strong substrate inhibition. 2,2',3-Trihydroxydiphenyl ether, catechol, 3-methylcatechol, and 4-methylcatechol were oxidized less efficiently and 3,4-dihydroxybiphenyl was oxidized considerably less efficiently. PMID- 8226679 TI - Identification of two components of the Serratia marcescens metalloprotease transporter: protease SM secretion in Escherichia coli is TolC dependent. AB - The Serratia marcescens metalloprotease (protease SM) belongs to a family of proteins secreted from gram-negative bacteria by a signal peptide-independent pathway which requires a specific transporter consisting of three proteins: two in the inner membrane and one in the outer membrane. The prtDSM and prtESM genes encoding the two S. marcescens inner membrane components were cloned and expressed in Escherichia coli. Their nucleotide sequence revealed high overall homology with the two analogous inner membrane components of the Erwinia chrysanthemi protease secretion apparatus and lower, but still significant, homology with the two analogous inner membrane components of the E. coli hemolysin transporter. When expressed in E. coli, these two proteins, PrtDSM and PrtESM, allowed the secretion of protease SM only in the presence of TolC protein, the outer membrane component of the hemolysin transporter. PMID- 8226680 TI - The cbb operons of the facultative chemoautotroph Alcaligenes eutrophus encode phosphoglycolate phosphatase. AB - The two highly homologous cbb operons of Alcaligenes eutrophus H16 that are located on the chromosome and on megaplasmid pHG1 contain genes encoding several enzymes of the Calvin carbon reduction cycle. Sequence analysis of a region from the promoter-distal part revealed two open reading frames, designated cbbT and cbbZ, at equivalent positions within the operons. Comparisons with known sequences suggested cbbT to encode transketolase (TK; EC 2.2.1.1) as an additional enzyme of the cycle. No significant overall sequence similarities were observed for cbbZ. Although both regions exhibited very high nucleotide identities, 93% (cbbZ) and 96% (cbbT), only the chromosomally encoded genes were heterologously expressed to high levels in Escherichia coli. The molecular masses of the observed gene products, CbbT (74 kDa) and CbbZ (24 kDa), correlated well with the values calculated on the basis of the sequence information. TK activities were strongly elevated in E. coli clones expressing cbbT, confirming the identity of the gene. Strains of E. coli harboring the chromosomal cbbZ gene showed high levels of activity of 2-phosphoglycolate phosphatase (PGP; EC 3.1.3.18), a key enzyme of glycolate metabolism in autotrophic organisms that is not present in wild-type E. coli. Derepression of the cbb operons during autotrophic growth resulted in considerably increased levels of TK activity and the appearance of PGP activity in A. eutrophus, although the pHG1-encoded cbbZ gene was apparently not expressed. To our knowledge, this study represents the first cloning and sequencing of a PGP gene from any organism. PMID- 8226681 TI - Multilevel regulation of the sporulation transcription factor sigma K in Bacillus subtilis. AB - Gene expression in the mother-cell compartment of the Bacillus subtilis sporangium is governed in part by the sporulation transcription factor sigma K. The production of sigma K is controlled at three levels: by a chromosomal rearrangement that generates the sigma K-coding sequence (sigK), by compartment specific transcription of sigK, and by conversion of the inactive pro-protein product of sigK (pro-sigma K) to sigma K. To investigate the function of these multiple levels of regulation, we constructed a set of strains that bypass the chromosomal rearrangement, pro-protein processing, or both levels of control. Here we show that one of the functions of the chromosomal rearrangement and pro protein processing is to prevent inappropriate production of sigma K under nonsporulation conditions. In the absence of both of these levels of control, a low level of sigma K-directed gene expression is observed during stationary phase after growth in rich medium. The appearance of sigma K under these conditions is probably due to a low level of sigma K-directed transcription from the sigK promoter in a positive feedback loop. We also report the construction of a strain that produces high levels of sigma K during growth. Using this strain, we demonstrate that the production of sigma K during growth is sufficient to induce a cascade of gene expression that closely mimics late events in the mother-cell line of gene expression. PMID- 8226682 TI - Regulation of the Bacillus subtilis acetate kinase gene by CcpA. AB - The Bacillus subtilis gene encoding acetate kinase was identified on the basis of sequence similarity to the Escherichia coli ackA gene and to a second E. coli gene closely related to ackA. Insertional inactivation of this region of the B. subtilis chromosome resulted in the disappearance of acetate kinase enzyme activity in cell extracts. The ackA gene was mapped to a site close to the ccpA gene, at 263 degrees. The transcriptional start site for B. subtilis ackA was located 90 bp upstream from the start of the coding region, and expression was increased by growth in the presence of excess glucose. Growth of the AckA- mutant was inhibited by glucose, suggesting that acetate kinase is important for excretion of excess carbohydrate. The stimulation of ackA expression by glucose was blocked in a CcpA- mutant, indicating that CcpA, which is required for glucose repression of certain carbon source utilization genes, including amyE, may also be involved in activation of carbon excretion pathways. Two sequences resembling the amyO operator site were identified upstream of the ackA promoter; removal of this region resulted in loss of glucose activation of ackA expression. PMID- 8226683 TI - A gene encoding arginyl-tRNA synthetase is located in the upstream region of the lysA gene in Brevibacterium lactofermentum: regulation of argS-lysA cluster expression by arginine. AB - The Brevibacterium lactofermentum argS gene, which encodes an arginyl-tRNA synthetase, was identified in the upstream region of the lysA gene. The cloned gene was sequenced; it encodes a 550-amino-acid protein with an M(r) of 59,797. The deduced amino acid sequence showed 28% identical and 49% similar residues when compared with the sequence of the Escherichia coli arginyl-tRNA synthetase. The B. lactofermentum enzyme showed the highly conserved motifs of class I aminoacyl-tRNA synthetases. Expression of the argS gene in B. lactofermentum and E. coli resulted in an increase in aminoacyl-tRNA synthetase activity, correlated with the presence in sodium dodecyl sulfate-polyacrylamide gels of a clear protein band that corresponds to this enzyme. One single transcript of about 3,000 nucleotides and corresponding to the B. lactofermentum argS-lysA operon was identified. The transcription of these genes is repressed by lysine and induced by arginine, showing an interesting pattern of biosynthetic interlock between the pathways of both amino acids in corynebacteria. PMID- 8226684 TI - Multiple antibiotic resistance in Pseudomonas aeruginosa: evidence for involvement of an efflux operon. AB - An outer membrane protein of 50 kDa (OprK) was overproduced in a siderophore deficient mutant of Pseudomonas aeruginosa capable of growth on iron-deficient minimal medium containing 2,2'-dipyridyl (0.5 mM). The expression of OprK in the mutant (strain K385) was associated with enhanced resistance to a number of antimicrobial agents, including ciprofloxacin, nalidixic acid, tetracycline, chloramphenicol, and streptonigrin. OprK was inducible in the parent strain by growth under severe iron limitation, as provided, for example, by the addition of dipyridyl or ZnSO4 to the growth medium. The gene encoding OprK (previously identified as ORFC) forms part of an operon composed of three genes (ORFABC) implicated in the secretion of the siderophore pyoverdine. Mutants defective in ORFA, ORFB, or ORFC exhibited enhanced susceptibility to tetracycline, chloramphenicol, ciprofloxacin, streptonigrin, and dipyridyl, consistent with a role for the ORFABC operon in multiple antibiotic resistance in P. aeruginosa. Sequence analysis of ORFC (oprK) revealed that its product is homologous to a class of outer membrane proteins involved in export. Similarly, the products of ORFA and ORFB exhibit homology to previously described bacterial export proteins located in the cytoplasmic membrane. These data suggest that ORFA-ORFB-oprK (ORFC)-dependent drug efflux contributes to multiple antibiotic resistance in P. aeruginosa. We propose, therefore, the designation mexAB (multiple efflux) for ORFAB. PMID- 8226685 TI - Novel mechanism for UV sensitivity and apparent UV nonmutability of recA432 mutants: persistent LexA cleavage following SOS induction. AB - The recA432 mutant allele was isolated (T. Kato and Y. Shinoura, Mol. Gen. Genet. 156:121-131, 1977) by virtue of its defect in cellular mutagenesis (Mut-) and its hypersensitivity to damage by UV irradiation (UVs), which were phenotypes expected for a recA mutant. However, we found that in a different genetic background (lexA51 sulA211 uvrB+), recA432 mutants expressed certain mutant phenotypes but not the Mut- and UVs phenotypes (D.G. Ennis, N. Ossanna, and D.W. Mount, J. Bacteriol. 171:2533-2541, 1989). We present several lines of evidence that these differences resulted from the sulA genotype of the cell and that the apparent UVs and Mut- phenotypes of the sulA+ derivatives resulted from lethal filamentation of induced cells because of persistent derepression of sulA. First, transduction of sulA(Def) mutations into the recA432 strains restored cellular mutagenesis and resistance to UV. Second, recA432 sulA+ strains underwent filamentous death following SOS-inducing treatments. Third, cleavage of LexA repressor in a recA432 strain continued at a rapid rate long after UV induction, at a time when cleavage of the repressor in the recA+ parental strain had substantially declined. Fourth, we confirmed that a single mutation (recA432) conferring both the UVs and Mut- phenotypes mapped to the recA gene. These findings indicate that the RecA432 mutant protein is defective in making the transition back to the deactivated state following SOS induction; thus, the SOS induced state of recA432 mutants is prolonged and can account for an excess of SulA protein, leading to filamentation. These results are discussed in the context of molecular models for RecA activation for LexA and UmuD cleavage and their roles in the control of mutagenesis and cell division in the SOS response. PMID- 8226686 TI - Functional domains of the penicillinase repressor of Bacillus licheniformis. AB - The penicillinase repressor (PENI) negatively regulates expression of the penicillinase gene (penP) in Bacillus licheniformis by binding to its operators located within the promoter region of penP.penI codes for a protein with 128 amino acids. Filter-binding analyses suggest that the active form of the repressor is a dimer. Genetic analyses of PENI derivatives showed that the repressor carrying either a 6-amino-acid deletion near the N terminus or a 14 amino-acid deletion at the C terminus was functionally inactive in vivo. A repressor derivative carrying a 6-amino-acid deletion within its N-terminal region was extensively purified and used in DNA footprinting and subunit cross linking analyses. The results of these studies showed that the repressor derivative had lost its ability to bind operator specifically even though it could dimerize effectively. In similar studies, we demonstrated that an N terminal portion of PENI with a molecular mass of 10 kDa derived by digestion with papain was able to bind operator specifically but with reduced affinity and had completely lost its ability to dimerize. These data suggest that the repressor has two functional and separable domains. The amino-terminal domain of the repressor is responsible for operator recognition, and the carboxyl-terminal domain is involved in subunit dimerization. PMID- 8226687 TI - Sequence analysis and interposon mutagenesis of the hupT gene, which encodes a sensor protein involved in repression of hydrogenase synthesis in Rhodobacter capsulatus. AB - The hupT gene, which represses hydrogenase gene expression in the purple photosynthetic bacterium Rhodobacter capsulatus, has been identified and sequenced. The nucleotide sequence of hupT and of the contiguous downstream open reading frame, hupU, is reported. The HupT protein of 456 amino acids (48,414 Da) has sequence similarity with the FixL, DctB, NtrB, and ArcB proteins and is predicted to be a soluble sensor kinase. Insertional inactivation of the hupT gene led to deregulation of transcriptional control, so that the hydrogenase structural operon hupSLC became overexpressed in cells grown anaerobically or aerobically. The HupT- mutants were complemented in trans by a plasmid containing an intact copy of the hupT gene. The hupU open reading frame, capable of encoding a protein of 84,879 Da, shared identity with [NiFe]hydrogenase subunits; the strongest similarity was observed with the periplasmic hydrogenase of Desulfovibrio baculatus. PMID- 8226688 TI - Hemin uptake in Porphyromonas gingivalis: Omp26 is a hemin-binding surface protein. AB - A 26-kDa outer membrane protein (Omp26) has been proposed to play a role in hemin acquisition by Porphyromonas gingivalis (T. E. Bramanti and S. C. Holt, J. Bacteriol. 174:5827-5839, 1992). We studied [55Fe]hemin uptake in P. gingivalis grown under conditions of hemin starvation (Omp26 expressed on the outer membrane surface) and hemin excess (Omp26 not expressed on surface). [55Fe]hemin uptake occurred rapidly in hemin-starved cells which incorporated up to 70% of total [55Fe]hemin within 3 min. P. gingivalis grown under hemin-starved conditions or treated with the iron chelator 2,2'-bipyridyl to induce an iron stress took up six times more [55Fe]hemin than hemin-excess-grown cells. Polyclonal monospecific anti-Omp26 antibody added to hemin-starved cells inhibited [55Fe]hemin uptake by more than 50%, whereas preimmune serum had no effect. [55Fe]hemin uptake in hemin starved P. gingivalis was inhibited (36 to 67%) in the presence of equimolar amounts of unlabeled hemin, protoporphyrin IX, zinz protoporphyrin, and Congo red dye but was not inhibited in the presence of non-hemin-containing iron sources. Heat shock treatment (45 degrees C) of hemin-excess-grown P. gingivalis (which cases translocation of Omp26 to the surface) increased [55Fe]hemin uptake by threefold after 3 min in comparison with cells grown at 37 degrees C. However, no [55Fe] hemin uptake beyond 3 min was observed in either hemin-excess-grown or hemin-starved cells exposed to heat shock. In experiments using heterobifunctional cross-linker analysis, hemin and selected porphyrins were cross-linked to Omp26 in hemin-starved P. gingivalis, but no cross-linking was seen with hemin-excess-grown cells. However, cross-linking of hemin to Omp26 was observed after heat shock treatment of hemin-excess-grown cells. Finally, anti Omp26 antibody inhibited cross-linked of hemin to Omp26. These findings indicate that hemin binding and transport into P.gingivalis cell mediated by Omp26. PMID- 8226689 TI - Cloning and molecular analysis of genes affecting expression of binding substance, the recipient-encoded receptor(s) mediating mating aggregate formation in Enterococcus faecalis. AB - Transfer of the conjugative plasmid pCF10 in Enterococcus faecalis strains involves production of a plasmid-encoded aggregation substance on the surface of donor cells in response to stimulation by a pheromone secreted by recipient cells. Aggregation substance then facilitates attachment to recipient cells via a chromosomally encoded receptor, termed binding substance (BS). A BS mutant, strain INY3000, generated by random Tn916 insertions, was previously found to carry copies of the transposon at four unique sites (K. M. Trotter and G. M. Dunny, Plasmid 24:57-67, 1990). In the present study, DNA flanking the Tn916 insertions was used to complement the BS mutation of INY3000 following Tn916 excision from cloned chromosomal fragments. Complementation results showed that three of the four regions mutated in INY3000 play some role in BS expression. Tn5 mutagenesis and DNA sequence analysis of the complementing fragment from one of these regions indicated the presence of three genes (ebsA, ebsB, and ebsC) that affect BS expression. The ebsA and ebsB genes encode peptides likely to function in cell wall metabolism, whereas ebsC may encode a product that suppresses the function or expression of EbsB. PMID- 8226690 TI - Biosynthesis of the corrin macrocycle of coenzyme B12 in Pseudomonas denitrificans. AB - Studies with cell-free protein preparations from a series of recombinant strains of Pseudomonas denitrificans demonstrated that precorrin-3 is converted into a further trimethylated intermediate, named precorrin-3B, along the pathway to coenzyme B12. It was then shown that the part of the pathway from precorrin-3 (called precorrin-3A hereafter) to precorrin-6x involves three intermediates, precorrin-3B, precorrin-4, and precorrin-5. Precorrin-3B was isolated in its native (reduced) as well as its oxidized (factor-IIIB) states, and precorrin-4 was isolated in its oxidized form only (factor-IV). Both factors were in vitro precursors of precorrin-6x. The synthesis of precorrin-6x from precorrin-3A was shown to be catalyzed by four enzymes, CobG, CobJ, CobM, and CobF, intervening in this order. They were purified to homogeneity. CobG, which converts precorrin-3A to precorrin-3B, was found to be an iron-sulfur protein responsible for the oxidation known to occur between precorrin-3A and precorrin-6x, and CobJ, CobM, and CobF are the C-17, C-11, and C-1 methylases, respectively. The acetate fragment is extruded after precorrin-4 formation. This study combined with our recent structural studies on factor-IV (D. Thibaut, L. Debussche, D. Frechet, F. Herman, M. Vuilhorgne, and F. Blanche, J. Chem. Soc. Chem. Commun. 1993:513-515, 1993) and precorrin-3B (L. Debussche, D. Thibaut, M. Danzer, F. Debu, D. Frechet, F. Herman, F. Blanche, and M. Vuilhorgne, J. Chem. Soc. Chem. Commun. 1993:1100 1103, 1993) provides a first step-by-step picture of the sequence of the enzymatic reactions leading to the corrin ring in P. denitrificans. PMID- 8226691 TI - The glnB region of the Escherichia coli chromosome. AB - We present sequences of the glnB gene of Escherichia coli and of two open reading frames (ORFs) located directly upstream of glnB and transcribed in the same direction. The major transcriptional start sites for glnB are located between ORF 2 and glnB, but some transcription of glnB is initiated at the promoter for ORF 1. The putative amino acid sequence of the ORF-2 product has high homology to that of response regulators which by phosphorylation acquire the ability to activate transcription of sigma 54-dependent promoters. The product of ORF-1 showed no similarity to other known proteins. The product of neither ORF-1 nor ORF-2 is necessary for the ability of PII, the product of glnB, to bring about the repression of glutamine synthetase in response to nitrogen excess. On the other hand, the product of hmpA, a gene located on the other side of glnB and transcribed in the opposite direction, appears to play an auxiliary role in this process. PMID- 8226692 TI - Fine mapping of the three rRNA operons on the updated genomic map of Campylobacter jejuni TGH9011 (ATCC 43431). AB - The three rRNA gene loci of Campylobacter jejuni TGH9011 (ATCC 43431) were cloned. All three rRNA operons were shown to possess a contiguous 16S-23S structure and contain intercistronic tRNA(Ala) and tRNA(Ile). The three RNA operons and additional 14 genetic markers were mapped in the updated genomic map of C. jejuni TGH9011, which now has a total of 24 genetic markers. PMID- 8226693 TI - Resistance of lambda cI translation to antibiotics that inhibit translation initiation. AB - The lambda cI lysogenic transcript is unusual in having no leader. Expression of a cI-lacZ protein fusion was relatively resistant to kasugamycin and pactamycin, which inhibit translation initiation on transcripts with leaders. Our data imply that there are distinct differences in translation initiation between the two classes of transcripts. PMID- 8226694 TI - Identification of the gene encoding an N-acetylpuromycin N-acetylhydrolase in the puromycin biosynthetic gene cluster from Streptomyces alboniger. AB - The biologically inactive compound N-acetylpuromycin is the last intermediate of the puromycin antibiotic biosynthetic pathway in Streptomyces alboniger. Culture filtrates from either this organism or Streptomyces lividans transformants harboring the puromycin biosynthetic gene cluster cloned in low-copy-number cosmids contained an enzymic activity which hydrolyzes N-acetylpuromycin to produce the active antibiotic. A gene encoding the deacetylase enzyme was located at one end of this cluster, subcloned in a 2.5-kb DNA fragment, and expressed from a high-copy-number plasmid in S. lividans. PMID- 8226695 TI - Heat shock and development induce synthesis of a low-molecular-weight stress responsive protein in the myxobacterium Stigmatella aurantiaca. AB - In the fruiting body-forming myxobacterium Stigmatella aurantiaca a 21,000-M(r) protein, SP21, is synthesized during fruiting, heat shock, and stress induced by oxygen limitation. The corresponding gene was isolated from a gene expression library in lambda gt11 with an antiserum to the purified protein. The DNA sequence of the gene reveals that SP21 is a member of the alpha-crystallin family of low-molecular-weight heat shock proteins. PMID- 8226696 TI - Molecular cloning and mapping of 16S-23S rRNA gene complexes of Staphylococcus aureus. AB - Staphylococcus aureus BB255, a derivative of NCTC8325, had six rRNA operons, and each operon contained two SmaI sites about 3 kb apart. By molecular cloning and pulsed-field gel electrophoresis, all operons were mapped at the junctions of SmaI fragments in the published map of NCTC8325 except one, which was connected to a previously unidentified 23-kb SmaI fragment. PMID- 8226697 TI - Lactose permease of Escherichia coli catalyzes active beta-galactoside transport in a gram-positive bacterium. AB - The following several lines of evidence demonstrate that lactose permease (LacY) of Escherichia coli is assembled into the cytoplasmic membrane of gram-positive Corynebacterium glutamicum, expressing the lacY gene, as a functional carrier protein. (i) LacY was detected immunologically in the cytoplasmic membrane fraction of the heterologous host. (ii) Recombinant C. glutamicum cells bearing the lacY gene displayed an increased influx of o-nitrophenyl-beta-D galactopyranoside, which was inhibited by N-ethylmaleimide. (iii) Washed cells were capable of accumulating methyl-beta-D-thiogalactoside about 60-fold. (iv) The uptake of methyl-beta-D-thiogalactoside was energy dependent and could be inhibited by the addition of 10 microM carbonyl cyanide-m-chlorophenylhydrazone. LacY of E. coli was active in the recombinant C. glutamicum cells despite the different membrane lipid compositions of these organisms. PMID- 8226698 TI - Negative autoregulation by the Escherichia coli SoxS protein: a dampening mechanism for the soxRS redox stress response. AB - The soxRS redox stress regulon of Escherichia coli is triggered in two stages, with the activated SoxR protein stimulating the soxS gene, whose product then triggers transcription of approximately 10 promoters. Genetic and biochemical experiments presented here show that SoxS protein also limits soxS transcription in vivo and binds the soxS promoter in vitro. PMID- 8226699 TI - Cloning, sequencing, and characterization of the gene encoding the class I fructose-1,6-bisphosphate aldolase of Staphylococcus carnosus. AB - fda from Staphylococcus carnosus TM300, encoding the class I fructose-1,6 bisphosphate aldolase, was cloned in Escherichia coli and sequenced. The 888 nucleotide open reading frame encoding a protein with an M(r) of 32,855 had an E. coli-like promoter sequence. Plasmids containing fda complemented E. coli NP315 (Fda-). Expression of fda in S. carnosus led to a six- to eightfold increase in aldolase production and activity; low levels of glucose in the growth medium stimulated activity. PMID- 8226700 TI - Site-directed mutagenesis reveals the importance of conserved charged residues for the transport activity of the PheP permease of Escherichia coli. AB - Site-directed mutagenesis has been used to identify a number of charged residues essential for the transport activity of the PheP protein. These residues are highly conserved in the cluster of amino acid transporters. However, some other conserved residues and a number of aromatic residues have been shown not to be essential for transport activity. PMID- 8226702 TI - Papers presented at the Symposium on Pathogenesis of HIV infection. Part 2. PMID- 8226701 TI - Analysis of the genetic requirements for viability of Escherichia coli K-12 DNA adenine methylase (dam) mutants. AB - RecBCD protein, necessary for Escherichia coli dam mutant viability, is directly required for DNA repair. Recombination genes recF+, recN+, recO+, and recQ+ are not essential for dam mutant viability; they are required for recBC sbcBC dam mutant survival. mutH, mutL, or mutS mutations do not suppress subinduction of SOS genes in dam mutants. PMID- 8226703 TI - Safety of the blood supply. AB - This is a review of events when the medical community realized that AIDS was an infectious disease which might be transmitted by blood transfusions and the response by the various organizations and agencies to curb the potential spread of HIV via blood products. It became possible through a number of approaches to make the blood supply safe so that today the likelihood of transmission of HIV by blood transfusion is extremely unlikely. PMID- 8226704 TI - Infection of hemophiliacs with HIV. AB - HIV was introduced into the blood supply at least 4 years before there was any clinical illness in hemophiliacs. While viral removal and inactivation techniques of plasma products have improved, contamination with a very stable virus is still a possibility. PMID- 8226705 TI - HIV infection in hemophilia. AB - This report reviews the natural history of hemophilia and the development of concentrates as the preferred form of treatment and discusses the nature of immune defect in persons with hemophilia. It examines the risk of hepatitis and effect of HIV infection on treatment protocols, and it surveys the status of the safety of modern concentrates and what we have to look forward to in the future. PMID- 8226706 TI - HIV inactivation in plasma products. AB - This is a review of therapeutic plasma products viz., coagulation factor concentrates, immunoglobulins, volume expanders, and protease inhibitors, and the progress made in rendering them free from transmitting infectious virus. Problems associated with the development of plasma products and the risks associated with their use are addressed. The recent technical advances designed to inactivate the hepatitis virus which led to the inactivation of HIV while preserving most of the biologic activity are reviewed. Today manufactured plasma products are essentially free from transmitting viral diseases of any significant clinical consequences. PMID- 8226707 TI - Evaluation of platelets collected by a new portable apheresis device. AB - We studied the efficiency of platelet collection by the Mobile Collection System (MCS) using two types of experimental protocols and evaluated the effect of storage at 22 degrees C on the platelet concentrates (PC). MCS is a new blood cell separator that combines discontinuous flow features with a new computerized operating system and can be used to harvest either full units of apheresis PC (SDP protocol) or half units of PC together with one to two units of plasma (PLP protocol). On the average, 1.98 x 10(11) +/- 0.46 x 10(11) (mean +/- SD) platelets were obtained by the PLP protocol and 3.01 x 10(11) +/- 0.70 x 10(11) and 4.2 x 10(11) +/- 1.12 x 10(11) by the early and later versions of the SDP protocols, respectively. The mean number of WBC per PC ranged from 3.3 to 4.7 x 10(8). During the storage period pH stayed above 7.0. On the average, the production of one molecule of lactate corresponded to the consumption of 0.538 molecules of glucose, indicating that less than 8% of glucose was consumed by the oxidative pathway. There were only small increases in LDH and B thromboglobulin concentrations. Furthermore, the ability of platelets to recover from osmotic shock and to aggregate following exposure to dual agonists declined only slightly during storage, indicating that both viability and function of platelets collected by the MCS were preserved during storage. PMID- 8226708 TI - Successful management of paraprotein-associated peripheral polyneuropathies by immunoadsorption of plasma with staphylococcal protein A. AB - Two patients with paraprotein-associated peripheral polyneuropathy were treated successfully using immunoadsorption of patient's plasma with staphylococcal protein A. Both had previously been treated with immunosuppressive agents or plasma exchange, and were rapidly relapsing at the time of their protein A immunoadsorption therapy. One patient was treated "on-line" with a blood cell separator, and one was treated "off-line." Both responded well to therapy with minimal toxicity. Serum levels of circulating immune complexes were elevated in one patient and remained so during and after therapy. Immunoadsorption with protein A should be investigated as a therapeutic option for patients with paraprotein-associated peripheral polyneuropathy. The therapy is relatively easy to administer, particularly "off-line," and was well tolerated by our patients. More experience, including formal clinical trials, will be required to properly define the indications for, and mechanism of response to, this therapy. PMID- 8226709 TI - Treatment of babesiosis by red blood cell exchange in an HIV-positive, splenectomized patient. AB - Babesiosis is a malaria-like parasitic disease causing subclinical or mild illness in most cases. Splenectomized patients, however, may experience a more severe course. Although generally responsive to antibiotic therapy, several cases of severe babesiosis refractory to appropriate antibiotic therapy have been reported to respond promptly and dramatically to red blood cell (RBC) exchange transfusion. Although the role of HIV coinfection in babesiosis is uncertain, two previously reported cases raise a concern that it may predispose to a more severe clinical course. We report a third case of severe babesiosis in an HIV-positive splenectomized man, following travel to an endemic area. Antibiotic therapy, though initially effective, ultimately failed to prevent severe disease. RBC exchange transfusion resulted in prompt clinical improvement, which has been sustained during 26 months of follow-up. Although the patient has since developed various sequelae of HIV infection, including disseminated Kaposi's sarcoma, CMV retinitis, and enteritis, there has been no recurrence of observable parasitemia. In severe babesiosis, RBC exchange transfusion, combined with appropriate antibiotic therapy, appears to be a rapidly effective therapeutic modality which can induce sustained remissions. PMID- 8226710 TI - Comparison of demographics and motivations of highly committed whole blood and platelet donors. AB - To compare donor demographics and motivations for donation, we surveyed 159 consecutive whole blood donors (WD) and 146 consecutive plateletpheresis donors. All donors had donation experiences of at least 2 gallons (i.e., 16 previous donations). Evaluation of answers to fixed-response and open-ended questions revealed no difference between the two groups in age, type of wages, compensation by employer, "time off" for donation, and church or volunteer organization memberships. The two groups differed in that the platelet donor (PD) group had a higher percentage of males than females (PD group 72.6% males and 27.4% females versus WD group 59% males and 41% females [chi-square = .01]). PDs most often began donating platelets because they were asked (22.6%), demonstrating the importance of active recruitment. When asked why they do not donate platelet products by apheresis, highly committed WDs reported lack of time (25%), lack of knowledge ("Don't know about it," 21%), lack of recruitment ("No one asked," 18.5%), or lack of awareness ("Never though about it," 5%) as reasons. Factors centering on time constraints, lack of knowledge or awareness about platelet donation by apheresis, and lack of active recruitment prohibit highly committed WDs from entering into plateletpheresis programs. PMID- 8226711 TI - Comparative bioavailability of ferric polymaltose and ferrous sulphate in iron deficient blood donors. AB - Absolute iron deficiency is treated by correcting the causative lesion and then, traditionally, administering sufficient amounts of ferrous salt to return the haemoglobin level to normal and replenish body stores. The bioavailability of ferric compounds has been questioned and accordingly their therapeutic role remains controversial. A special problem is posed by regular blood donation, where the frequency of phlebotomy is limited by the haemoglobin level, which, in turn, requires maintenance of an adequate supply of iron from dietary sources. Since this latter situation may not always occur, it would be of practical benefit to have a form of supplementation that is effective and can be taken without side effects. These issues were prospectively examined in a consecutive series of otherwise healthy blood donors who developed absolute iron deficiency anaemia and were then randomly allocated to receive 60 mg of this metal as ferrous sulphate twice a day (Group 1: n = 51), 100 mg as chewable ferric polymaltose daily (Group 2: n = 53), or the latter product twice a day (Group 3: n = 55). Serial studies showed that 80% of patients in Groups 1 and 3 had reached normal haemoglobin levels by 12 weeks, but this figure was only 50% in Group 2. Similarly, the proportion of patients improving their percentage saturation of transferrin to within the normal range was significantly better in Groups 1 and 3 than in Group 2 (P < .01). However, body iron stores, reflected in serum ferritin level, was significantly better in Group 1 (P < .01); there was no difference in this respect between Groups 2 and 3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8226712 TI - Summary of results of 1991 ASFA apheresis survey. American Society for Apheresis. AB - As part of gathering data for its Networking Directory, the Registry/Directory Committee of the American Society for Apheresis solicited data on the types of apheresis procedures performed, equipment used, and diseases treated in the year 1991. In this voluntary solicitation of the 266 centers responding, 94% were from the United States. For those centers reporting exact numbers of procedures performed out of all institutions doing the procedures 48,221 therapeutic and 330,702 donor procedures were reported. The dominant type of equipment being used is the centrifugal type. For therapeutic procedures plasma exchange is the most prevalent procedure and for donor procedures it is plateletpheresis. For therapeutics, Guillain-Barre syndrome was the most prevalent disease treated by plasmapheresis and leukemias by cytapheresis. The survey methodology proved to be valuable in gathering the data on a voluntary basis. With subsequent surveys and refinement of its methodology the apheresis practices in the United States could be assessed quantitatively to provide information on the number and types of procedures being carried out. Such information is required to assess the impact of apheresis technology on health care, to project trends, and to review the cost impact of this technology. PMID- 8226713 TI - NADH-quinone oxidoreductase, the most complex complex. PMID- 8226714 TI - Attempts to define distinct parts of NADH:ubiquinone oxidoreductase (complex I). AB - The NADH:ubiquinone oxidoreductase (complex I) is made up of a peripheral part and a membrane part. The two parts are arranged perpendicular to each other and give the complex an unusual L-shaped structure. The peripheral part protrudes into the matrix space and constitutes the proximal segment of the electron pathway with the NADH-binding site, the FMN and at least three iron-sulfur clusters. The membrane part constitutes the distal segment of the electron pathway with at least one iron-sulfur cluster and the ubiquinone-binding site. Both parts are assembled separately and relationships of the major structural modules of the two parts with different bacterial enzymes suggest, that both parts also emerged independently in evolution. This assumption is further supported by the conserved order of bacterial complex I genes, which correlates with the topological arrangement of the corresponding subunits in the two parts of complex I. PMID- 8226715 TI - Characteristics of the energy-transducing NADH-quinone oxidoreductase of Paracoccus denitrificans as revealed by biochemical, biophysical, and molecular biological approaches. AB - A comparison of the mitochondrial NADH-ubiquinone oxidoreductase and the energy transducing NADH-quinone oxidoreductase (NDH-1) of Paracoccus denitrificans revealed that both systems have similar electron-transfer and energy-transduction pathways. In addition, both complexes are sensitive to the same inhibitors and contain similar electron carriers, suggesting that the Paracoccus NDH-1 may serve as a useful model system for the study of the human enzyme complex. The gene cluster encoding the Paracoccus NDH-1 has been cloned and sequenced. It is composed of 18,106 base pairs and contains 14 structural genes and six unidentified reading frames (URFs). The structural genes, URFs, and their polypeptides have been characterized. We also discuss nucleotide sequences which are believed to play a role in the regulation of the NDH-1 gene cluster and Paracoccus NDH-1 subunits which may contain the binding sites of substrates and/or electron carriers. PMID- 8226716 TI - Bacterial NADH-quinone oxidoreductases: iron-sulfur clusters and related problems. AB - Many bacteria contain proton-translocating membrane-bound NADH-quinone oxidoreductases (NDH-1), which demonstrate significant genetic, spectral, and kinetic similarity with their mitochondrial counterparts. This review is devoted to the comparative aspects of the iron-sulfur cluster composition of NDH-1 from the most well-studied bacterial systems to date.: Paracoccus denitrificans, Rhodobacter sphaeroides, Escherichia coli, and Thermus thermophilus. These bacterial systems provide useful models for the study of coupling Site I and contain all the essential parts of the electron-transfer and proton-translocating machinery of their eukaryotic counterparts. PMID- 8226717 TI - The proton-translocating NADH: ubiquinone oxidoreductase: a discussion of selected topics. AB - The proton-translocating NADH:ubiquinone oxidoreductase (complex I) is a large, multi-subunit and multi-redox centre enzyme which is found in the mitochondrial inner membrane and plasma membrane of some bacteria. In this minireview an attempt has been made to critically discuss selected topics in the structure and function of this enzyme. A special emphasis is given to the iron-sulphur cluster and to the proteins that may bind them. Previous suggestions for the mechanism of proton-translocation by complex I are discussed. Subcomplexes that contain several but not all of the subunits of the intact mitochondrial enzyme are described and analysed in order to identify the functional core of the enzyme. The data on the trans-membrane organisation of several subunits is examined. It is hoped that most of the suggestions as well as the questions raised here could be experimentally tested in the near future. PMID- 8226718 TI - Kinetics, control, and mechanism of ubiquinone reduction by the mammalian respiratory chain-linked NADH-ubiquinone reductase. AB - In mammalian cells the membrane-bound NADH-quinone oxidoreductase serves as the entry point for oxidation of NADH in the respiratory chain and as the proton translocating unit which conserves the free energy of the enzyme intramolecular redox reactions as the free energy of the electrochemical proton gradient across the coupling membrane. This review summarizes the kinetic properties of the mammalian enzyme. Emphasis is placed on the hysteretic properties of the enzyme as related to the possible control of intramitochondrial NADH oxidation and to the mechanism of the enzyme interaction with ubiquinone. Recent evidence for participation of flavin and the protein-bound ubisemiquinone pair in the enzyme catalyzed proton translocation mechanism are discussed. PMID- 8226719 TI - NAD(P)H-ubiquinone oxidoreductases in plant mitochondria. AB - Plant (and fungal) mitochondria contain multiple NAD(P)H dehydrogenases in the inner membrane all of which are connected to the respiratory chain via ubiquinone. On the outer surface, facing the intermembrane space and the cytoplasm, NADH and NADPH are oxidized by what is probably a single low-molecular weight, nonproton-pumping, unspecific rotenone-insensitive NAD(P)H dehydrogenase. Exogenous NADH oxidation is completely dependent on the presence of free Ca2+ with a K0.5 of about 1 microM. On the inner surface facing the matrix there are two dehydrogenases: (1) the proton-pumping rotenone-sensitive multisubunit Complex I with properties similar to those of Complex I in mammalian and fungal mitochondria. (2) a rotenone-insensitive NAD(P)H dehydrogenase with equal activity with NADH and NADPH and no proton-pumping activity. The NADPH-oxidizing activity of this enzyme is completely dependent on Ca2+ with a K0.5 of 3 microM. The enzyme consists of a single subunit of 26 kDa and has a native size of 76 kDa, which means that it may form a trimer. PMID- 8226720 TI - Na(+)-translocating NADH-quinone reductase of marine and halophilic bacteria. AB - The respiratory chain of marine and moderately halophilic bacteria requires Na+ for maximum activity, and the site of Na(+)-dependent activation is located in the NADH-quinone reductase segment. The Na(+)-dependent NADH-quinone reductase purified from marine bacterium Vibrio alginolyticus is composed of three subunits, alpha, beta, and gamma, with apparent M(r) of 52, 46, and 32 kDa, respectively. The FAD-containing beta-subunit reacts with NADH and reduces ubiquinone-1 (Q-1) by a one-electron transfer pathway to produce ubisemiquinones. In the presence of the FMN-containing alpha-subunit and the gamma-subunit, Q-1 is converted to ubiquinol-1 without the accumulation of free radicals. The reaction catalyzed by the alpha-subunit is strictly dependent on Na+ and is strongly inhibited by 2-n-heptyl-4-hydroxyquinoline N-oxide (HQNO), which is tightly coupled to the electrogenic extrusion of Na+. A similar type of Na(+) translocating NADH-quinone reductase is widely distributed among marine and moderately halophilic bacteria. The respiratory chain of V. alginolyticus contains another NADH-quinone reductase which is Na+ independent and has no energy-transducing capacity. These two types of NADH-quinone reductase are quite different with respect to their mode of quinone reduction and their sensitivity toward NADH preincubation. PMID- 8226721 TI - Ferricytochrome c induces monophasic kinetics of ferrocytochrome c oxidation in cytochrome c oxidase. AB - The kinetics of ferrocytochrome c oxidation by reconstituted cytochrome c oxidase (COX) from bovine heart was followed by a spectrophotometric method, using on line data collection and subsequent calculation of reaction rates from a function fitted to the progress curve. When reaction rates were calculated at increasing reaction times, the multiphasic kinetics of ferrocytochrome c oxidation gradually changed into monophasic Michaelis-Menten kinetics. The same phenomenon was observed when ferrocytochrome c oxidation was followed in the presence of increasing amounts of ferricytochrome c. From these results we conclude that ferricytochrome c shifts the multiphasic kinetics of ferrocytochrome c oxidation by COX into monophasic kinetics, comparable to high ionic strength conditions. Furthermore, we show that ferricytochrome c inhibits the "high affinity phase" of ferrocytochrome c oxidation in an apparently competitive way, while inhibition of the "low affinity phase" is noncompetitive. These findings are consistent with a "regulatory site model" where both the catalytic and the regulatory site bind ferro- as well as ferricytochrome c. PMID- 8226722 TI - Subunit structures of purified beef mitochondrial cytochrome bc1 complex from liver and heart. AB - The existence of tissue-specific isozymes of cytochrome c oxidase has been widely documented. We have now studied if there are differences between subunits of mitochondrial bc1 complexes isolated from liver and heart. For this purpose, we have developed a method for the purification of an active ubiquinol-cytochrome c oxidoreductase from adult bovine liver that includes solubilization of submitochondrial particles with deoxycholate, ammonium acetate fractionation, resolubilization with dodecyl maltoside, and ion exchange chromatography. The electrophoretic pattern of the liver preparation showed the presence of 11 subunits, with apparent molecular weights identical to the ones reported for the heart complex. Western blot analysis and isoelectric focusing followed by two dimensional gels of bc1 complexes from liver and heart were compared, and no qualitative differences were observed. In addition, the high-molecular-weight subunits of the purified complexes from both tissues, subunits I, II, V, and VI, were isolated by PAGE in the presence of Coomasie Blue and subjected to limited proteolysis and to chemical digestion with cyanogen bromide and BNPS-skatol, and the peptide patterns were compared. Finally, two of the small-molecular-weight subunits from the liver complex were isolated (subunits VII and X), partially analyzed by amino terminal sequencing, and found to be identical with the reported sequence of their heart counterparts. The data suggest that, in contrast to the case of cytochrome c oxidase, bc1 complexes from liver and heart do not exhibit tissue-specific differences. PMID- 8226723 TI - NADH-ascorbate free radical and -ferricyanide reductase activities represent different levels of plasma membrane electron transport. AB - Plasma membranes isolated from rat liver by two-phase partition exhibited dehydrogenase activities for ascorbate free radical (AFR) and ferricyanide reduction in a ratio of specific activities of 1:40. NADH-AFR reductase could not be solubilized by detergents from plasma membrane fractions. NADH-AFR reductase was inhibited in both clathrin-depleted membrane and membranes incubated with anti-clathrin antiserum. This activity was reconstituted in plasma membranes in proportion to the amount of clathrin-enriched supernatant added. NADH ferricyanide reductase was unaffected by both clathrin-depletion and antibody incubation and was fully solubilized by detergents. Also, wheat germ agglutinin only inhibited NADH-AFR reductase. The findings suggest that NADH-AFR reductase and NADH-ferricyanide reductase activities of plasma membrane represent different levels of the electron transport chain. The inability of the NADH-AFR reductase to survive detergent solubilization might indicate the involvement of more than one protein in the electron transport from NADH to the AFR but not to ferricyanide. PMID- 8226724 TI - Gal4 protein binding is required but not sufficient for derepression and induction of GAL2 expression. AB - The Saccharomyces cerevisiae GAL2 gene upstream activator sequence (UAS) region was examined for protein bound in vivo by chromatin footprinting at high resolution. Gal4 transcriptional activator protein binds to the two consensus UAS sites whether GAL2 expression is induced, uninduced, or repressed by growth with different carbon sources. Although wild type strains show loss of the Gal4 protein-specific footprint in repressing media containing glucose, constitutive high level expression of Gal4 protein restores the GAL2 UAS footprints without fully derepressing GAL2 transcription. Thus binding of the Gal4 activator to target sites in the DNA is required but not sufficient for GAL2 derepression and induction. Gal4-independent protein-DNA complexes were also detected in the region, including one over the previously noted centromere-binding protein (CP1) site upstream of the Gal4 complexes. PMID- 8226725 TI - Translational control by influenza virus. Selective translation is mediated by sequences within the viral mRNA 5'-untranslated region. AB - In cells infected by influenza virus type A, host cell protein synthesis declines rapidly and dramatically, while influenza viral protein synthesis occurs efficiently throughout infection. Previously, we had shown that the selective translation of influenza viral mRNAs in infected cells occurred in a cap dependent manner and was due at least in part to structures inherent in the mRNAs. Using chimeras containing the noncoding and coding regions of cellular and viral mRNAs, we can now report that the selective translation is mediated by sequences within the 5'-untranslated regions (UTR) of the viral mRNAs. Polysome analysis confirmed that a 45-nucleotide sequence contained in the 5'-UTR of the influenza viral nucleocapsid protein was necessary and sufficient to allow the host cell translational machinery to discriminate between viral and cellular mRNAs. In reciprocal experiments in which the 5'-UTR of the cellular mRNA secreted embryonic alkaline phosphatase replaced the nucleocapsid protein 5'-UTR, viral protein synthesis was inhibited in virus-infected cells, resembling host protein synthesis. Finally, we demonstrated that the 5'-UTR of another influenza viral mRNA, that encoding the nonstructural protein, also conferred resistance to the shutoff of protein synthesis in influenza virus-infected cells. PMID- 8226726 TI - Functional expression of insulin receptor substrate-1 is required for insulin stimulated mitogenic signaling. AB - To examine the role of the insulin receptor substrate-1 (IRS-1) in mediating insulin biological responsiveness, we generated Chinese hamster ovary cell lines expressing antisense IRS-1 RNA. These cells displayed morphological alterations as well as markedly reduced growth rates compared to the parental cells. Furthermore, the antisense IRS-1 cell lines had decreased insulin-stimulated IRS 1 tyrosine phosphorylation, reduced phosphatidylinositol 3-kinase activation, and decreased thymidine incorporation relative to the parental cell line. Insulin dependent transcriptional regulation of a serum response element/luciferase reporter construct (SRE-Luc) was also reduced in the antisense IRS-1-expressing cell lines. However, co-transfection with a plasmid directing the expression of rat IRS-1 fully restored insulin-stimulated SRE-Luc activity in the IRS-1 antisense cell lines. Thus, the inhibition in insulin signaling was a specific effect of decreased IRS-1 tyrosine phosphorylation. Taken together, these data demonstrate that insulin regulation of mitogenic signaling requires the functional expression of IRS-1 and documents its central importance in the insulin intracellular signaling pathway. PMID- 8226727 TI - Gi-mediated activation of the p21ras-mitogen-activated protein kinase pathway by alpha 2-adrenergic receptors expressed in fibroblasts. AB - The alpha 2-adrenergic receptors are linked to inhibition of adenylylcyclase and, under certain circumstances, to stimulation of phospholipid hydrolysis via pertussis toxin-sensitive G proteins. Here we show that alpha 2-adrenergic receptors can couple to an alternative signaling pathway. When expressed in Rat-1 cells, stimulation of the alpha 2A receptor, which couples to Gi2 and Gi3, causes rapid, transient activation of the protooncogene product p21ras as measured by an increase in the amount of bound GTP. Furthermore, alpha 2A receptor stimulation causes rapid phosphorylation of the p42 mitogen-activated protein (MAP) kinase. Pertussis toxin completely inhibits both p21ras activation and MAP kinase phosphorylation, but both responses appear to be independent of adenylylcyclase inhibition or phospholipase stimulation. Thus, alpha 2-adrenergic receptors can couple to the p21ras-MAP kinase pathway via Gi, which may explain the mitogenic potential of alpha 2 agonists in certain cell types; together with previous results, these findings further suggest that activation of this pivotal signaling pathway may be a common event in the action of Gi-coupled receptors. PMID- 8226728 TI - Adipose cell hyperplasia and enhanced glucose disposal in transgenic mice overexpressing GLUT4 selectively in adipose tissue. AB - To gain insight into the molecular pathogenesis of obesity and specifically the role of nutrient partitioning in the development of obesity, we overexpressed the insulin-responsive glucose transporter (GLUT4) in transgenic mice under the control of the fat-specific aP2 fatty acid-binding protein promoter/enhancer. Two lines of transgenic mice were generated, which overexpressed GLUT4 6-9-fold in white fat and 3-5-fold in brown fat with no overexpression in other tissues. In vivo glucose tolerance was enhanced in transgenic mice. In isolated epididymal, parametrial, and subcutaneous adipose cells from transgenic mice, basal glucose transport was 20-34-fold greater than in nontransgenic littermates. Insulin stimulated glucose transport was 2-4-fold greater in cells from transgenic mice. Total body lipid was increased 2-3-fold in transgenic mice overexpressing GLUT4 in fat. Surprisingly, fat cell size was unaltered and fat cell number was increased > 2-fold. This is the first animal model in which increased fat mass results solely from adipocyte hyperplasia and it will be a valuable model for understanding the mechanisms responsible for fat cell replication and/or differentiation in vivo. PMID- 8226729 TI - Regulation of the GTPase cycle of the neuronally expressed Ras-like GTP-binding protein Rab3A. AB - The small GTP-binding protein Rab3A (identical to smg p25A) is expressed in neural/exocrine/endocrine cells, is distributed between the cytosol and secretory vesicle membranes, and may cycle between these locations to regulate exocytosis. It is proposed that the GTP/GDP state of Rab3A controls this distribution. In PC12 cells, cytosolic Rab3A is predominantly GDP-bound, whereas membrane associated Rab3A is approximately 50% GTP-bound. Two cytosolic factors, GDP dissociation inhibitor (GDI) and guanine nucleotide releasing factor (GRF), act only on GDP.Rab3A, and preferentially with post-translationally modified Rab3A. Rab3A GTPase-activating protein (GAP) does not preferentially act on processed Rab3A, and interacts selectively with GTP.Rab3A. GDI antagonizes GRF but not GAP activity toward Rab3A. These data are consistent with the concept of an ordered Rab3A cycle controlled by factors that regulate the guanine-nucleotide binding state of Rab3A. PMID- 8226730 TI - Activation of platelet phosphatidylinositide 3-kinase requires the small GTP binding protein Rho. AB - The small GTP-binding protein Rho regulates the assembly of actin stress fibers and focal adhesions in cells responding to growth factors. ADP-ribosylation of Rho by C3 transferase blocks this function; however, an enzymatic target for Rho has not yet been defined. We now report that Rho activates phosphatidylinositide 3-kinase in soluble preparations of platelets. Activation of phosphatidylinositide 3-kinase by GTP gamma S is blocked by ADP-ribosylation of endogenous Rho, and Rho shifts to the cytoskeleton in platelets exposed to thrombin. The inhibitory effects of ADP-ribosylation are overcome by exogenous recombinant Rho but not by recombinant Rac, another member of the Ras superfamily. Exposure of platelets to thrombin has been reported to lead to activation of phosphatidylinositide 3-kinase, a shift of this enzyme to the platelet membrane skeleton, and rapid cytoskeletal reorganization. In other studies, ADP-ribosylation of Rho has been found to inhibit thrombin-induced platelet aggregation, a cytoskeletally linked event. We suggest that Rho may exert its effects on cytoskeletal reorganization via phosphatidylinositide 3 kinase. PMID- 8226731 TI - Rab3A GTPase-activating protein-inhibiting activity of Rabphilin-3A, a putative Rab3A target protein. AB - Rabphilin-3A is a putative target protein for Rab3A, a member of the small G protein superfamily that is implicated in regulated secretion, particularly in neurotransmitter release. Rabphilin-3A contains at least two functionally different domains: the N-terminal Rab3A-binding domain and the C-terminal C2 domain, which interacts with both Ca2+ and phospholipid. Because Rabphilin-3A interacts preferentially with GTP-Rab3A rather than with GDP-Rab3A, we have examined here whether Rabphilin-3A affects the GTPase activity of Rab3A. Rabphilin-3A and its N-terminal fragment, but not its C-terminal fragment, very weakly stimulated the basal GTPase activity of Rab3A. However, Rabphilin-3A and its N-terminal fragment strongly inhibited the Rab3A GAP-stimulated GTPase activity of Rab3A. Ca2+ and phospholipid showed no effect on these activities of Rabphilin-3A. The physiological significance of the GAP activity of Rabphilin-3A is obscure, but it is likely that Rabphilin-3A inhibits Rab3A GAP activity and keeps Rab3A in the GTP-bound active form during its action as a target molecule for Rab3A. PMID- 8226732 TI - The role of endoplasmic reticulum calcium pumps during cytosolic calcium spiking in pancreatic acinar cells. AB - Many cell types show repetitive short lasting cytosolic calcium spikes with long interspike periods when stimulated with submaximal concentrations of agonists linked to the phosphoinositide signaling pathway. In pancreatic acinar cells these spikes have been shown to be evoked by constant levels of inositol trisphosphate through a mechanism of calcium-induced calcium release and do not depend acutely on the presence of external calcium. However, the processes involved in the interspike period have remained unclear. Here we report that the endoplasmic reticulum Ca(2+)-ATPase play a significant role, not only in resequestering calcium after a spike, but also in regulating the long interspike period. Decreasing the activity of the endoplasmic reticulum calcium pumps leads to shorter interspike intervals and thus higher spiking frequencies, while the duration of each spike increases. The endoplasmic reticulum Ca(2+)-ATPases are able to entirely suppress a response that can subsequently be evoked by partial inhibition of the pumps. This suggests that during the interspike period there is a considerable amount of calcium released from intracellular stores, which is rapidly buffered by the endoplasmic reticulum calcium pumps and the cytosolic calcium-binding proteins. A calcium spike will be initiated by calcium-induced calcium release only when the buffering is saturated. PMID- 8226733 TI - Oscillations of secretion driven by oscillations of cytoplasmic Ca2+ as evidences in single pancreatic islets. AB - It is often assumed, but has not been demonstrated, that the oscillations of cytoplasmic calcium (Ca2+i) that occur in various secretory cells induce oscillations in secretion. Here, we have used single pancreatic islets from normal mice to monitor simultaneously insulin release and Ca2+i in beta-cells for periods up to 25 min. Ca2+i and insulin secretion were found to oscillate in synchrony during stimulation by glucose. This synchrony persisted when the frequency of both events changed spontaneously or upon addition of the hypoglycaemic sulfonylurea tolbutamide, or when their relative amplitudes varied. Repolarizing the beta-cell membrane with diazoxide abolished both Ca2+i and insulin oscillations. In contrast, epinephrine suppressed insulin oscillations in spite of the persistence of Ca2+i oscillations. This study is the first direct demonstration that, in an intact organ, sustained oscillations of Ca2+i induced by a physiological stimulus entrain synchronous oscillations of the functional response, and that both events can vary simultaneously or be dissociated depending on the experimental conditions. PMID- 8226734 TI - A novel intersubunit repair mechanism in the tryptophan synthase alpha 2 beta 2 complex. Critical role of the beta subunit lysine 167 in intersubunit communication. AB - This study explores intersubunit communication in the tryptophan synthase alpha 2 beta 2 complex from Salmonella typhimurium. We find that an engineered mutation in the contact region between the alpha and beta subunits remarkably alters the catalytic and spectroscopic properties of the beta subunit in the alpha 2 beta 2 complex. Ligands that bind to the alpha subunit largely repair the deleterious effects of the beta subunit mutation in the alpha 2 beta 2 complex. The conserved residue chosen for mutation, beta subunit lysine 167, appears to form an ion pair with alpha subunit aspartate 56 in the crystal structure of the wild type alpha 2 beta 2 complex. Although replacement of beta subunit lysine 167 by threonine does not prevent formation of the alpha 2 beta 2 complex, this mutation reduces the rate of synthesis of L-tryptophan from L-serine and indole (beta reaction) 25 fold. Ligands that bind to the alpha subunit (indole-3-glycerol phosphate, indole 3-propanol phosphate, alpha-glycerol 3-phosphate, or potassium phosphate) largely restore the activity of the mutant alpha 2 beta 2 complex in the beta reaction. We conclude that beta subunit lysine 167 plays an important role in intersubunit communication in the alpha 2 beta 2 complex. The striking allosteric effects of alpha subunit ligands on the mutant beta subunit in the alpha 2 beta 2 complex may result from ligand-induced conformational changes in the alpha subunit that are transmitted to the beta subunit and repair the mutational defect in the beta subunit. PMID- 8226735 TI - Hydrophobic amino acid in the i2 loop plays a key role in receptor-G protein coupling. AB - Signal transduction of the heptahelical G protein-coupled receptors (GPCRs) involves multiple receptor domains, but a universal consensus domain for coupling has not yet been defined. Alanine mutagenesis scanning was performed on the intracellular loops and the COOH tail of the human muscarinic cholinergic receptor (Hm1) to identify coupling domains. Stimulation of phosphatidylinositol (PI) turnover was determined after transfection of the alanine mutants into U293 human embryonic kidney cells. Alanine substitutions in four regions (loops i1, i2, and NH2 and COOH junctions of i3) impaired coupling efficiency by approximately 50% or more, but the strongest reduction (> 80%) resulted from alanine replacement of a single amino acid, leucine 131. This residue is located in the middle of the second intracellular loop (i2), within the highly conserved GPCR motif (DRYXXV(I)XXPL). The position equivalent to Leu-131 in Hm1 contains a bulky hydrophobic amino acid (L, I, V, M, or F) in nearly all cloned GPCRs. Substitution of Leu-131 with polar amino acids (aspartate and asparagine) also resulted in strongly defective coupling, whereas phenylalanine (found in the equivalent position in the beta 2 adrenoceptor) can replace leucine without losing PI coupling ability of Hm1. Alanine substitution of the corresponding amino acid in the Hm3 receptor (L174A) also inhibited agonist-stimulated PI turnover, while replacing Phe-139 with alanine in the beta 2 adrenoceptor suppressed stimulation of adenylyl cyclase. We propose that a bulky hydrophobic amino acid in the middle of the i2 loop serves as a general site relevant to G protein coupling, whereas coupling selectivity is governed by other receptor domains. PMID- 8226736 TI - Protein kinase C modulation of fibronectin matrix assembly. AB - Fibroblasts have cell surface sites that mediate the assembly of fibronectin (Fn) into the extracellular matrix. Treatment of fibroblasts with kinase inhibitors (ML-7, H7, HA1004, calphostin C, and staurosporine) resulted in the rapid decrease in the binding of 125I-labeled plasma Fn and iodinated amino-terminal fragments of Fn. The dose responses of the four inhibitors suggest that the target kinase is protein kinase C (PKC) rather than the cyclic AMP- or cyclic GMP dependent kinases. Three different fibroblastic cells were similarly affected. The inhibition was rapid and reversible and could not be overcome by increasing concentrations of Fn. Treatment of fibroblasts with phorbol esters and other agents that activate PKC resulted in increased amounts of 125I-labeled Fn binding to the cell surface. These results imply that Fn matrix assembly is modulated by PKC-mediated phosphorylation. PMID- 8226737 TI - Refolding of the precursor and mature forms of mitochondrial aspartate aminotransferase after guanidine hydrochloride denaturation. AB - The mitochondrial isozyme of aspartate aminotransferase (mAspAT), a dimeric pyridoxal phosphate (PLP)-dependent enzyme, is encoded by the nuclear genome and synthesized in the cytoplasm as a precursor protein (pmAspAT) containing a 29 residue amino-terminal signal peptide which is essential for its targeting and import into mitochondria. In the cytosolic-like environment of rabbit reticulocyte lysate, newly synthesized rat liver pmAspAT has been found to slowly fold and bind PLP (Mattingly, J. R., Jr., Youssef, J., Iriarte, A. and Martinez Carrion, M. (1993) J. Biol. Chem. 268, 3925-3937). On the other hand, isolated mammalian (pig) mAspAT, when denatured with guanidine hydrochloride, seems unable to refold to a catalytically active state (West, S. M., and Price, N. C. (1990) Biochem. J. 265, 45-50). With the availability of rat liver recombinant precursor and mature forms of mAspAT as homogeneous, stable preparations, an assessment of the influence of the signal peptide on the in vitro refolding of this protein can be made. Following unfolding induced by guanidine hydrochloride, we have investigated the refolding process of this complex, dimeric coenzyme-dependent protein system by activity, fluorescence, and circular dichroism. Both mAspAT and pmAspAT can be efficiently renatured after rapid dilution of the denaturing agent at low protein concentrations. The equilibrium unfolding/refolding transitions and the kinetics of folding are protein concentration-independent and identical for both protein forms. Binding of coenzyme into the active site pocket seems to occur at a late step in the folding process of both mAspAT and pmAspAT, suggesting that in these proteins the coenzyme does not direct the folding of the polypeptide chain. These results indicate that the in vitro refolding of mAspAT is not regulated or influenced by the presence of the amino-terminal signal peptide. On the other hand, in vitro refolding in buffer is significantly faster than the folding of newly synthesized precursor protein in reticulocyte lysate examined in our previous report (reference above), pointing at the likely influence of cytosolic factors in modulating folding in the cell. PMID- 8226738 TI - Chemical and kinetic evidence for an essential histidine residue in the electron transfer from aromatic donor to horseradish peroxidase compound I. AB - Horseradish peroxidase, when incubated with diethyl pyrocarbonate (DEPC), a histidine-specific reagent, shows time-dependent inactivation to oxidize aromatic electron donor, guaiacol. The inactivation follows pseudo-first order kinetics with a second order rate constant of 0.67 min-1 M-1. The pH dependence of inactivation shows an inflection point at 6.02, indicating histidine derivatization by DEPC. A difference spectrum of modified versus native enzyme shows a peak at 244 nm for N-carbethoxyhistidine that is diminished by hydroxylamine. Stoichiometric studies indicate that out of 2 histidine residues modified, one is responsible for inactivation. A plot of log reciprocal half-time of inactivation against log DEPC concentration suggests that only 1 histidine is essential. From the computer-stimulated structure of horseradish peroxidase, we tentatively suggest that this critical histidine is most likely distal histidine 42. Binding studies show that this histidine is not involved in guaiacol binding. Modified enzyme forms compound I with H2O2 but not compound II, suggesting a block of electron transfer process. Modified compound I cannot oxidize guaiacol as evidenced by the absence of donor-induced spectral shift from 408 nm, suggesting a block of electron transfer from bound donor to compound I. We suggest that this tentatively identified distal histidine controls aromatic donor oxidation by regulating electron transport without affecting donor binding or compound I formation. PMID- 8226739 TI - Expression of endogenous NMDAR1 transcripts without receptor protein suggests post-transcriptional control in PC12 cells. AB - Expression of RNA for the NMDAR1 subunit of the N-methyl-D-aspartate receptor was detected by Northern hybridization in both nerve growth factor-differentiated and undifferentiated rat pheochromocytoma (PC12) cells. The NMDA receptor type 1 (NMDAR1) message in PC12 cells was similar in size to that expressed in hippocampal neurons. PC12 cell cDNAs that were amplified by polymerase chain reaction with primers flanking the coding region of NMDAR1 corresponded to the NMDAR1 splice variant NMDA receptor type 1 isoform C (NMDAR1C). Using calcium imaging or patch-clamp recording, no functional NMDA-gated ion channels were found in PC12 cells. A monoclonal antibody against NMDAR1 was developed in order to investigate whether or not NMDAR1 protein was present in PC12 cells. Only trace amounts of NMDAR1 protein were found in native PC12 cells. However, expression of NMDAR1 protein was detected in PC12 cells that were transfected with an expression vector containing an NMDAR1C clone under control of a cytomegalovirus promoter. These findings suggest that the expression of NMDAR1 protein in PC12 cells may be controlled by post-transcriptional mechanisms. The PC12 cell line may serve as a model system for the study of the transcriptional, post-transcriptional, and translational regulation of NMDAR1. Furthermore, the presence of NMDAR1 RNA in a particular cell type may not necessarily indicate expression of NMDAR1 protein. PMID- 8226740 TI - Multiple cytosolic calcium signals and membrane electrical events evoked in single arginine vasopressin-stimulated corticotrophs. AB - The action of arginine vasopressin (AVP) on cytosolic free Ca2+ concentration ([Ca2+]i) was investigated in single rat pituitary corticotrophs using indo-1 microfluorimetry, in part in combination with the monitoring of membrane electrical events with the perforated patch-clamp technique. In corticotrophs showing the series of short-lived [Ca2+]i rises (transient pattern) in response to corticotropin-releasing factor, 100 nM AVP evoked either the transient pattern or a [Ca2+]i spike followed by a sustained plateau (spike/plateau pattern). Not all corticotrophs responded to changes in AVP concentration in the same manner. Some cells exhibited a concentration-dependent increase in [Ca2+]i transient activity, whereas others showing the spike/plateau at high AVP concentrations responded to low agonist concentrations by two [Ca2+]i responses: a slow rising step or two to three sinusoidal-like oscillations. Combined [Ca2+]i and patch clamp recordings as well as manipulation of extracellular Ca2+ showed that both transient pattern and the plateau of spike/plateau response depended on Ca2+ entry mainly through voltage-gated, dihydropyridine-sensitive Ca2+ channels. By contrast, step, oscillations, and spike were due to Ca2+ release from internal stores. These Ca(2+)-mobilizing responses caused the activation of Ca(2+) activated, apamin-sensitive K+ channels, which led to a membrane hyperpolarization. These results reveal cell-specific [Ca2+]i signals and associated electrical events in individual AVP-stimulated corticotrophs. PMID- 8226741 TI - Mechanisms of camptothecin resistance in yeast DNA topoisomerase I mutants. AB - The anti-cancer drug camptothecin targets eukaryotic DNA topoisomerase I by trapping the covalent complex formed between the catalytically active enzyme and DNA. Saccharomyces cerevisiae cells expressing yeast DNA topoisomerase I mutant top1 vac (I725R, N726A) or top1N726L, in which the amino acid residues N terminus to the active site tyrosine Tyr-727 were changed as indicated, were found to be camptothecin-resistant even though the mutant proteins expressed in Escherichia coli were previously shown to be active. Assays of enzyme-catalyzed relaxation of supercoiled DNA in vitro and in vivo in yeast showed that the camptothecin resistance of these mutants arises by entirely different mechanisms. Top1N726L catalyzed DNA relaxation was not detected in yeast. The Top1 vac protein was catalytically active; however, camptothecin was inefficient in trapping the covalent intermediate formed between the Top1 vac enzyme and DNA. Yeast cells expressing human mutant htop1 vac, with similar substitutions near the active site tyrosine Tyr-723, were also camptothecin-resistant. Surprisingly, in the absence of camptothecin, yeast rad52 mutants defective in the repair of double stranded DNA breaks were nonviable when top1N726L or top1 vac was overexpressed but viable when htop1 vac was overexpressed. These results suggest differences between yeast and human enzyme function in vivo. PMID- 8226742 TI - The GTPase-activating NF1 fragment of 91 amino acids reverses v-Ha-Ras-induced malignant phenotype. AB - The human neurofibromatosis type 1 gene encodes a Ras GAP (GTPase-activating protein) of 2818 amino acids called NF1. This GAP contains a domain of 338 amino acids (residues 1194-1531) called NF1-GRD, which shares 26% sequence identity with the C-terminal domain (GAP1C, residues 709-1044) of another Ras GAP called GAP1. Both NF1-GRD and GAP1C activate normal Ras GTPases but not oncogenic mutants such as v-Ha-Ras. Any attempt to reverse the v-Ha-Ras-induced malignant transformation by the GAP1C has, so far, been unsuccessful. However, we have found that when the NF1-GRD is overexpressed in v-Ha-Ras-transformed NIH/3T3 cells, it greatly reduces their ability to form colonies in a soft agar, the property tightly associated with their malignancy. This is, so far, the first demonstration that a Ras-binding protein can act as a potent antagonist of the oncogenic Ras mutants in mammalian cells. In an attempt to further screen the smallest anti-oncogenic fragment derived from the NF1-GRD, we have prepared a series of its deletion mutants and examined their interaction with Ras first by monitoring their GAP activity (ability to activate the normal Ras GTPase). The deletion analysis has revealed that the N-terminal 247 amino acids (residues 1194 1440) of NF1-GRD are not required for its GAP activity, suggesting that its remaining domain of 91 amino acids (NF91, residues 1441-1531) is sufficient to bind the v-Ha-Ras, although its GAP activity is 20 times lower than the NF1-GRD. The NF91 is indeed able to reverse the v-Ha-Ras-induced malignant transformation as the NF1-GRD. The NF91 is, so far, the smallest among the tumor suppressor proteins that show the anti-v-Ha-Ras action in vivo. Thus, the NF91 appears to be a good starting material from which a smaller and more potent v-Ha-Ras antagonist could be devised to be used as a potential cure for the human cancers caused by the Ras mutants. PMID- 8226743 TI - Mutational analysis of the cation-independent mannose 6-phosphate/insulin-like growth factor II receptor. A consensus casein kinase II site followed by 2 leucines near the carboxyl terminus is important for intracellular targeting of lysosomal enzymes. AB - The cation-independent mannose 6-phosphate/insulin-like growth factor II receptor (CI-MPR) mediates intracellular sorting of lysosomal enzymes, binding lysosomal enzymes in the Golgi and delivering them to a lysosomal compartment. The receptor also mediates endocytosis of extracellular ligands. We have devised a new method that rigorously measures function of the CI-MPR in intracellular sorting and used it to identify a previously uncharacterized signal near the COOH terminus of the receptor that is required for sorting. We stably transfect mutant receptors into CI-MPR-deficient mouse L cells, isolate homogeneous clonal cell lines that express a range of receptor levels for each mutant, and assay each cell line for levels of receptor expression and secretion of total phosphorylated lysosomal enzymes. Examination of the secretion phenotype of the cells as a function of receptor levels provides a sensitive indicator of the intrinsic sorting efficiency of each mutant receptor. We find that chimeric CI-MPRs that contain the bovine extracytoplasmic domain and the human or mouse transmembrane and cytoplasmic domains function identically to the bovine receptor, thus demonstrating that sorting signals are conserved. Analysis of a series of truncation and alanine scanning mutants reveals that a consensus casein kinase II site followed by 2 leucines near the COOH terminus that has the sequence ( 10)DDSDEDLL(-3) is important for receptor function in sorting of lysosomal enzymes. PMID- 8226744 TI - Expression of the extracellular domain of the rat liver prolactin receptor and its interaction with ovine prolactin. AB - A clone of the extracellular domain of the rat liver prolactin receptor was generated by the RNA-based polymerase chain reaction, and the NH2-terminal 210 amino acids were expressed in HeLa cells using a vaccinia virus/T7 hybrid expression system. The protein was isolated from serum-free culture medium directly by chromatography on an ovine prolactin affinity column and yielded approximately 1.5 mg of protein/liter of suspension culture. The extracellular domain of the rat prolactin receptor inhibited the ovine prolactin-dependent mitogenesis of rat lymphoma Nb2 cells with an IC50 of 7.1 pM and bound 125I labeled ovine prolactin with a Kd of 1.21 +/- 0.19 nM. In contrast, the binding of the 125I-labeled extracellular domain to ovine prolactin exhibited positive cooperativity with a Hill coefficient of 1.73. High pressure gel filtration chromatography was used to demonstrate the formation of a complex consisting of one molecule of ovine prolactin and two molecules of the extracellular domain of the rat prolactin receptor. Complex formation occurred with human growth hormone, but not with ovine growth hormone, a non-lactogen. PMID- 8226745 TI - Identification of the essential cysteine residue in the active site of bovine pyruvate dehydrogenase. AB - Pyruvate dehydrogenase (E1), the first catalytic component of the bovine pyruvate dehydrogenase complex, is composed of two nonidentical subunits in a tetrameric alpha 2 beta 2 form. The sulfhydryl-specific reagent N-ethylmaleimide (NEM) was used to identify the reactivities and function of cysteinyl residues and subsequent identification of these residues in the active site of bovine E1. Treatment of E1 with 0.2 mM NEM resulted in loss (90%) of enzymatic activity; the inactivation followed bimolecular reaction kinetics. The inactivation was almost entirely prevented by thiamin pyrophosphate (TPP) and pyruvate; protection is probably due to formation of the hydroxyethylidene-TPP intermediate. To identify the reactive cysteinyl residues in the active site region, the nonessential SH groups in E1 were first modified with NEM in the presence of TPP and pyruvate. After quenching with dithiothreitol and removal of the substrate and cofactor by dialysis, the modified E1 was treated with [14C]NEM to label the exposed cysteinyl residue(s) in or near the active site region. The data indicate that NEM reacted in the active site region of the E1 component with a stoichiometry of 2 mol of [14C]NEM bound per mol of E1 tetramer. The initial rapid labeling of E1 with [14C]NEM established that incorporation was predominantly into the alpha subunit. A single radiolabeled peptide was isolated following V8 protease digestion of radiolabeled E1 by [14C]NEM. Sequence analysis of the labeled peptide derived from bovine E1 demonstrated that the labeled cysteinyl residue was equivalent to Cys-62 in the alpha subunit (mature form) of human E1. PMID- 8226746 TI - Rat liver heme oxygenase. High level expression of a truncated soluble form and nature of the meso-hydroxylating species. AB - A rat heme oxygenase (HO-1) gene without the sequence coding for the last 23 amino acids has been constructed and expressed behind the pho A promoter in Escherichia coli. The enzyme is expressed at high levels as a soluble catalytically active protein that causes the bacterial cells to accumulate biliverdin. The purified truncated heme-heme oxygenase complex is spectroscopically indistinguishable from the complex with the native enzyme and converts heme to biliverdin when reconstituted with rat liver cytochrome P450 reductase. Reaction of the recombinant heme-heme oxygenase complex with H2O2 produces a species with the spectroscopic properties of verdoheme. Unidentified products are obtained when this intermediate is directly extracted from the protein, but biliverdin is obtained if the verdoheme-protein complex is exposed to cytochrome P450 reductase and NADPH before the extraction step. In contrast, reaction of the heme-heme oxygenase complex with meta-chloroperbenzoic acid (mCPBA), tert-butylhydroperoxide, or cumene hydroperoxide yields a ferryl (FeIV = O) complex. Reaction of the heme-heme oxygenase complex with mCPBA also produces an EPR-detectable protein radical. In accord with formation of a ferryl intermediate, recombinant heme oxygenase catalyzes the mCPBA- and alkylhydroperoxide-dependent peroxidation of 2-methoxyphenol (guaiacol). Guaiacol oxidation is not observed during turnover of the enzyme by cytochrome P450 reductase/NADPH or H2O2. Conversely, biliverdin is not formed with tert butylhydroperoxide or mCPBA. H2O2 thus supports the first step of the normal catalytic oxidation of heme by heme oxygenase, but alkyl and acyl hydroperoxides do not. These results suggest that the alpha-meso-hydroxylation required for biliverdin formation is mediated by the distal of the two oxygens in the iron dioxygen intermediate (Fe-O-O) engendered by reaction with either cytochrome P450 reductase/NADPH or H2O2. PMID- 8226747 TI - Regulation of thymidine kinase and thymidylate synthase in intact human lymphoblast CCRF-CEM cells. AB - It has been reported that thymidylate synthase (TS) is a component of a multienzyme complex associated with DNA replication based on observations that enzyme activity was decreased when cells were treated with various DNA synthesis inhibitors (Plucinski, T. M., Fager, R. S., and Reddy, G. P. V. (1990) Mol. Pharmacol. 38, 114-120). The veracity of the TS assay (known as the tritium release assay) employed in these experiments may be compromised, however, because it requires the S phase-specific enzyme thymidine kinase (TK) to phosphorylate the substrate [5-3H]dUrd. In our study, this problem was further illustrated as the phosphorylated products of [14C]dCyd and [6-3H]dUrd were simultaneously quantitated to determine the activities of TS, TK, and dCyd kinase in intact CCRF CEM cells. TS and dCyd kinase were unaffected by aphidicolin, hydroxyurea, and 1 beta-D-arabinofuranosylcytosine, whereas TK was strongly inhibited by these agents. Elevation of the cellular dTTP pool that accompanied drug treatment was not the primary mechanism affecting TK activity because incubation of cells with dCyd elevated the dTTP pool to similar levels, but did not inhibit TK to the same extent as did the drugs. Furthermore, after cells were washed from aphidicolin, [6-3H]dCyd incorporation, which primarily labels dTMP in DNA, proceeded at a linear rate, whereas a lag period of 15 min was observed before [3H]dThd was incorporated at a linear rate. These results suggest that TK activity is affected by more than one mechanism in intact cells. Because the activities of dCyd kinase and dCMP deaminase do not fluctuate as much as that of TK in response to changes in DNA synthesis activity and cell cycle, dCyd incorporation appears to be a more reliable assay of TS in intact cells than does dUrd incorporation. Our findings also imply that [3H]dThd incorporation assays may overestimate inhibition of DNA synthesis. PMID- 8226748 TI - The inactivation of Fe-S cluster containing hydro-lyases by superoxide. AB - We report in this paper that highly purified Escherichia coli dihydroxy-acid dehydratase, fumarase A, fumarase B, and mammalian aconitase are inactivated by O2- with second order rate constants in the range of 10(6) to 10(7) M-1 s-1. Each of these enzymes belongs to the hydro-lyase class and contains catalytically active [4Fe-4S] clusters. Simultaneous with inactivation by O2- is the release of iron from their clusters. Our working hypothesis is O2- inactivates these enzymes by oxidizing their clusters to an unstable oxidation state, and cluster degradation follows. Consistent with this hypothesis is our observation that spinach dihydroxy-acid dehydratase, a member of the hydro-lyase class that has a catalytically active [2Fe-2S] cluster, is not inactivated and does not lose iron in the presence of O2-. Porcine fumarase, a member of the hydro-lyase class that does not contain an Fe-S cluster, is also not inactivated by O2-. We also report the rate constants for the inactivation of E. coli dihydroxy-acid dehydratase, fumarase A, fumarase B, and mammalian aconitase by O2 are close to 2 x 10(2) M-1 s-1, and the rate constants for the inactivation of E. coli dihydroxy-acid dehydratase and mammalian aconitase by H2O2 are about 10(3) M-1 s-1. E. coli dihydroxy-acid dehydratase has been reported previously to be inactivated in vivo when cells are grown in hyperbaric O2, presumably due to the increased O2- generated under these conditions. We report here that E. coli fumarase A, fumarase B, and aconitase are also inactivated in vivo by hyperbaric O2. Thermodynamic parameters for the oxidation of the cluster of aconitase by O2- and O2 are calculated. PMID- 8226749 TI - Stimulated Ca2+ influx raises mitochondrial free Ca2+ to supramicromolar levels in a pancreatic beta-cell line. Possible role in glucose and agonist-induced insulin secretion. AB - The effects of stimulated Ca2+ influx on cytosolic ([Ca2+]c) or intramitochondrial free Ca2+ ([Ca2+]m) were examined in the new pancreatic beta cell line, INS-1. [Ca2+]c was monitored by video imaging of single fura-2-loaded INS-1 cells, or in populations of cells transfected with non-targeted (cytosolic) aequorin. [Ca2+]m was measured after transfection with aequorin targeted to the mitochondria by fusion of the gene in frame with the signal peptide of cytochrome c oxidase subunit VIII. Two physiological stimuli of native beta-cells, glucose and ATP, raised [Ca2+]c in INS-1 cells largely by stimulating Ca2+ influx. Thus, glucose (20 mM) induced repetitive transient increases in [Ca2+]c (0.42 min-1, mean amplitude 229 nM above 102 nM basal). These transients were largely due to periodic stimulation of Ca2+ influx through voltage-sensitive Ca2+ channels, since they could be rapidly and reversibly blocked by chelation of external Ca2+, by addition of the hyperpolarizing agent diazoxide, or with the Ca2+ channel blocker SR 7037. ATP, by contrast, caused single transient [Ca2+]c increases, to about 300 nM above basal levels, which could be inhibited by > 90% upon external Ca2+ chelation. Challenge of aequorin-transfected cells with ATP increased [Ca2+]m to 4 microM or above, an effect blocked by EGTA. Furthermore, plasma membrane depolarization with high K+, used as a glucose surrogate to mimic, in a synchronized fashion, the influx-induced Ca2+ transients observed at the single cell level, also increased [Ca2+]m to > 4 microM. Similar increases in [Ca2+]m were also measured in other aequorin-transfected insulin-secreting cells, RINm5F, during mobilization of internal Ca2+ with carbachol. In contrast, glucose-induced changes in [Ca2+]m were below the level of detection in INS-1 cell populations, consistent with the asynchrony of the [Ca2+]c transients induced by this nutrient at the single-cell level, and the consequent small average [Ca2+]c rise. These data are in line with the view that stimulated Ca2+ influx into excitable cells raises [Ca2+]m as efficiently as internal Ca2+ mobilization in nonexcitable cells. In the case of INS-1 and pancreatic beta-cells, this may be important both to enhance oxidative metabolism, hence fueling the secretory process, and also to maintain the production of metabolic signaling molecules. PMID- 8226750 TI - Determination of the side chain pKa values of the lysine residues in calmodulin. AB - The 7 Lys residues in mammalian calmodulin (CaM) were reductively methylated with 13C-enriched formaldehyde and studied by (1H,13C)-heteronuclear multiple quantum coherence (HMQC) NMR. The apo- and Ca(2+)-forms of CaM, as well as a complex with a 22-residue peptide which comprises the CaM binding region of myosin light chain kinase were studied. The complete assignment of the two-dimensional NMR spectra was obtained by site-directed mutagenesis (Lys-->Gln) of all the Lys. The pKa values for the individual Lys could be determined by pH titration experiments. In Ca(2+)-CaM, the pKa values range from 9.29 (Lys-75) to 10.23 (Lys-77). The Lys in apo-CaM have higher pKa values than those in Ca(2+)-CaM. The binding of the myosin light chain kinase peptide gives rise to an increase of the pKa values of Lys-148 and Lys-75 by 0.5 and 0.8 pH units, respectively; this results from the relocation of their side chains to a completely solvent accessible state. The changes in the pKa values upon binding Ca2+ or the myosin light chain kinase peptide show a remarkable correlation with earlier reported chemical reactivity changes. Thus, our results indicate that pKa values, rather than structural and steric effects, play the dominant role in determining the reactivity of Lys side chains towards small electrophilic chemical modification reagents. The methodology used here could prove useful for the determination of individual pKa values in other proteins. PMID- 8226751 TI - Trapoxin, an antitumor cyclic tetrapeptide, is an irreversible inhibitor of mammalian histone deacetylase. AB - Trapoxin (cyclo-(L-phenylalanyl-L-phenylalanyl-D-pipecolinyl-L-2-amino-8- oxo 9,10-epoxy-decanoyl)) is a fungal product that induces morphological reversion from transformed to normal in sis-transformed NIH3T3 fibroblasts. Trapoxin was found to cause accumulation of highly acetylated core histones in a variety of mammalian cell lines. In vitro experiments using partially purified mouse histone deacetylase showed that a low concentration of trapoxin irreversibly inhibited deacetylation of acetylated histone molecules. Chemical reduction of an epoxide group in trapoxin completely abolished the inhibitory activity, suggesting that trapoxin binds covalently to the histone deacetylase via the epoxide. In contrast, inhibition by trichostatin A, a known potent inhibitor of histone deacetylase, was reversible. Despite the different mode of inhibition, trapoxin and trichostatin A induced almost the same biological effects on the cell cycle and differentiation. These results strongly suggest that the in vivo effects commonly induced by these agents can be attributed to histone hyperacetylation resulting from the inhibition of histone deacetylase. PMID- 8226752 TI - The role of COOH-terminal and acidic domains in the activity and stability of human insulin receptor protein tyrosine kinase studied by purified deletion mutants of the beta subunit domain. AB - We have previously expressed the human insulin receptor beta subunit domain containing transmembrane and cytoplasmic domains (IRTMTPK) in insect cells, and showed that the purified IRTMTPK was highly active (Li, S. L., Yan, P.-F., Pax, I. B., and Fujita-Yamaguchi, Y. (1992) Biochemistry 31, 12455-12462). To investigate the role of COOH-terminal and acidic domains of the insulin receptor kinase, we have expressed deletion mutants IRTMTPK delta CT (delta 76 amino acids) and IRTMTPK delta Acid (delta 19 amino acids). Both enzymes were purified by a one-step method using the same immunoaffinity column as used for IRTMTPK. While Km and Vmax for prephosphorylated IRTMTPK and delta Acid mutant enzyme determined using poly(Glu, Tyr)(4:1) were similar, catalytic efficiency of the delta CT mutant enzyme was significantly lower than those of IRTMTPK and delta Acid mutant enzyme as judged by Km and Vmax. Experiments for thermostability and susceptibility to proteases revealed that Tm of delta CT mutant enzyme was 3.5 degrees C lower than that of IRTMTPK enzyme (= 33.3 degrees C) and that delta CT mutant enzyme was digested by either trypsin or Lys-C into a 28,000 core domain much faster than IRTMTPK. Activation of delta CT mutant enzyme by polylysine was less significant than that of IRTMTPK and delta Acid mutant enzyme, approximately 4-versus approximately 17-fold. These studies suggested that the COOH-terminal domain plays important roles in both catalytic efficiency and stability of the insulin receptor kinase, and that the acidic domain by itself is not responsible for kinase activation by polylysine. PMID- 8226753 TI - Radiation-activated DNA-binding protein constitutively present in ataxia telangiectasia nuclei. AB - We have recently described the appearance of a specific DNA-binding protein in nuclei from human cells exposed to ionizing radiation which was not detected in nuclear extracts from unperturbed cells (Singh, S. P., and Lavin, M. F. (1990) Mol. Cell. Biol. 10, 5279-5285). We report here a similar activity which is constitutively present in nuclei of both unirradiated and irradiated cells from patients with the human genetic disorder ataxia telangiectasia (A-T). Activity was present in unirradiated nuclear extracts from 3 A-T cell lines of different complementation groups, but was not detected or was present only at a low level in 4 controls. Active protein was detected in the cytoplasm of both cell types. Exposure to ionizing radiation did not change the amount of DNA binding activity in A-T nuclei but led to an increase in nuclei from 4 control cell lines. Purification of the binding activities from A-T nuclei and control cytoplasm was carried out by affinity chromatography, as described previously for control extracts (Teale, B., Singh, S. P., Khanna, K. K., Findik, D., and Lavin, M. F. (1992) J. Biol. Chem. 267, 10295-10301). Southwestern analysis and UV cross linking confirmed the presence of a major DNA-binding species at 70 kDa in both cases with a minor binding activity at 47 kDa also evident. It was not possible to distinguish between the binding activities from A-T and control cells under different conditions, and phosphorylation was required for binding activity in both cases. Footprint analysis revealed that the same sequence was being recognized by the control and A-T proteins. The constitutive presence of a specific radiation-responsive DNA-binding protein in A-T cells may be indicative of a continuous state of stress in these cells. PMID- 8226754 TI - beta-Lapachone, a novel DNA topoisomerase I inhibitor with a mode of action different from camptothecin. AB - beta-Lapachone is a plant product that has been found to have many pharmacological effects. To date, very little is known about its biochemical target. In this study, we found that beta-lapachone inhibits the catalytic activity of topoisomerase I from calf thymus and human cells. But, unlike camptothecin, beta-lapachone does not stabilize the cleavable complex, indicating a different mechanism of action. beta-Lapachone inhibits topoisomerase I-mediated DNA cleavage induced by camptothecin. Incubation of topoisomerase I with beta lapachone before adding DNA substrate dramatically increases this inhibition. Incubation of topoisomerase I with DNA prior to beta-lapachone makes the enzyme refractory, and treatment of DNA with beta-lapachone before topoisomerase has no effect. These results suggest a direct interaction of beta-lapachone with topoisomerase I rather than DNA substrate. beta-Lapachone does not inhibit binding of enzyme to DNA substrate. In cells, beta-lapachone itself does not induce a SDS-K(+)-precipitable complex, but it inhibits complex formation with camptothecin. We propose that the direct interaction of beta-lapachone with topoisomerase I does not affect the assembly of the enzyme-DNA complex but does inhibit the formation of cleavable complex. PMID- 8226755 TI - Membrane topology of a P-type ATPase. The MgtB magnesium transport protein of Salmonella typhimurium. AB - P-type ATPases are a family of cation transport enzymes present in all species from bacteria to mammals whose members mediate membrane flux of all common biologically relevant cations. More than 50 members of this family of transporters have been sequenced; extensive structural data are available, and several members have been analyzed by site-directed mutagenesis. Nonetheless, there is no current consensus regarding their membrane topology. In this work, the Salmonella typhimurium Mg2+ transporting P-type ATPase encoded by the MgtB locus has been used as a model for P-type ATPases. Unlike other prokaryotic P type ATPases, the MgtB protein is similar in length, amino acid sequence, and hydropathy profile to known eukaryotic P-type ATPases. The membrane topology of MgtB was analyzed by several epitope insertions in MgtB and from the activity of 35 protein fusions between MgtB and the reporter enzymes BlaM (beta-lactamase) and LacZ (beta-galactosidase). The epitope insertions within MgtB all retained function as assessed by cation uptake assays and were regulated normally by the level of Mg2+ within the growth medium. The epitope insertion and fusion protein data are completely incompatible with the numerous previously proposed models for P-type ATPases predicting 7, 8, 9, or 12 transmembrane segments. Rather, they indicate that MgtB contains 10 transmembrane segments with both amino and carboxyl termini residing within the cytosol. By extension, we suggest that all eukaryotic P-type ATPases contain 10 transmembrane segments with both termini within the cytosol. PMID- 8226756 TI - Site-directed mutagenesis of human thioredoxin. Identification of cysteine 74 as critical to its function in the "early pregnancy factor" system. AB - Thioredoxin has been identified as a key component of the "early pregnancy factor" system, a system of components present in pregnancy sera which expresses a lymphocyte modifying activity in an assay known as the rosette inhibition assay. Although thioredoxin alone is inactive, addition of thioredoxin to lymphocytes in combination with nonpregnancy sera or platelet-activating factor results in a positive response. We have changed several amino acids of human thioredoxin by site-directed mutagenesis to investigate the residues required for this cooperative function. Conversion of the two active site residues (cysteines 32 and 35) to serines results in a protein devoid of classical redox activity; however, this protein retained its ability to cooperate with non-pregnancy sera or platelet-activating factor in the rosette inhibition assay. Vertebrate thioredoxins contain an additional conserved pair of cysteine residues in the C terminal portion of the protein. Changing both to serines resulted in no change in redox activity but completely abolished function in the rosette inhibition assay. Further study revealed this function was solely dependent on cysteine 74 as conversion of only cysteine 74 to serine abolished function, whereas replacement of only cysteine 70 with serine had no effect. The nonfunctional mutants counteracted the action of pregnancy serum in the assay strongly supporting the hypothesis that thioredoxin is an integral part of the early pregnancy factor system with residue cysteine 74 having an important role. PMID- 8226757 TI - The 2.0-A resolution structure of Escherichia coli histidine-containing phosphocarrier protein HPr. A redetermination. AB - The x-ray structure of Escherichia coli HPr has been redetermined at 2.0-A resolution. In contrast to the previous study (El-Kabbani, O. A. L., Waygood, E. B., and Delbaere, L. T. J. (1987) J. Biol. Chem. 262, 12926-12929), the overall structure is, in general, similar to other reported NMR and x-ray HPr structures, although there are some important differences in detail. The overall folding topology of HPr is a classical open-faced beta-sandwich, consisting of four antiparallel beta-strands and three alpha-helices. The least square refinement produced an R index of 0.135 for all measured unique data between 8.0 and 2.0 A resolution. The active center consists of His15 which is hydrogen bonded to a sulfate anion, and Arg17 which has a fully open conformation. This corresponds to the first observed "semi-closed" conformation of the active center of HPr. The Streptococcus faecalis HPr structure (Jia, Z., Vandonselaar, M., Quail, J. W., and Delbaere, L. T. J. (1993) Nature 361, 94-97) has the "open" conformation in which the side chains of His15 and Arg17 are directed as far away from each other as possible. The Bacillus subtilis HPr (Herzberg, O., Reddy, P., Sutrina, S., Saier, M. H., Jr., Reizer, J., and Kapadia, G. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 2499-2503) has the "closed" conformation in which the side chains of His15 and Arg17 are close together with a sulfate anion located in the active center. The open conformation represents the unphosphorylated form of HPr whereas the closed conformation likely resembles the phosphorylated form of HPr. The semi closed conformation observed in the E. coli HPr structure could represent a structural intermediate on the phosphorylation/dephosphorylation pathway of HPr. PMID- 8226758 TI - ATP hydrolysis is not stoichiometrically linked with proteolysis in the ATP dependent protease La from Escherichia coli. AB - Protein degradation in Escherichia coli involves the ATP-dependent serine protease La. Protease La is a homotetramer with one proteolytic and one ATP binding site per monomer. Its proteolytic activity has been shown to be highly increased by simultaneous hydrolysis of ATP, which is essential for the degradation of protein substrates by this enzyme. We have cloned and purified a proteolytically inactive La mutant, in which the catalytically active serine residue at position 679 was replaced by alanine. Fluorescence and circular dichroism spectra of the purified wild type and mutant enzyme revealed identical conformations of the proteins. Based on this observation, the catalytic properties of the wild type enzyme and the S679A mutant were compared. Although the S679A mutant lacks proteolytic activity toward both peptide and protein substrates under all conditions investigated, its ATPase activity is completely unaffected by the removal of the protease activity. Since protein substrates stimulate the ATP-dependent hydrolysis of peptides by protease La, it has been argued that this stimulation is due to interactions with a regulatory binding site on the enzyme. In accordance with this model, protein substrates such as alpha-casein and denatured bovine serum albumin stimulate the ATPase activity of the S679A mutant to the same degree as in the active protease. Therefore, the intrinsic ATPase activity of protease La as well as its stimulation is not dependent on the simultaneous hydrolysis of the protein substrate. PMID- 8226759 TI - Construction, expression, and characterization of a mutated animal fatty acid synthase deficient in the dehydrase function. AB - The location of the dehydrase domain in the multifunctional animal fatty acid synthase has been determined by engineering a fatty acid synthase mutant deficient in dehydrase activity. A full-length fatty acid synthase cDNA encoding a mutated histidine residue (His878-->Ala) was constructed and expressed in insect Sf9 cells using a baculoviral vector. The mutated recombinant fatty acid synthase retained all partial activities of the multifunctional complex except the dehydrase and was unable to synthesize fatty acids. beta-Hydroxybutyryl moieties were formed by the mutant fatty acid synthase from acetyl-CoA, malonyl CoA, and NADPH and slowly released as the CoA thioester, confirming that this protein cannot perform the dehydration reaction. This finding points to an important catalytic role for His878 in the dehydration reaction and establishes that the dehydrase domain is located immediately adjacent to the carboxyl terminus of the transferase domain. Examination of the completed domain map for the animal fatty acid synthase indicates that the catalytic domains are clustered in two groups separated by a central structural core: the ketoacyl synthase, malonyl/acetyltransferase, and dehydrase in the amino-terminal half and the enoyl reductase, ketoreductase, acyl carrier protein, and thioesterase in the carboxyl terminal half. A model is proposed in which the two centers for acyl chain initiation, elongation and termination, are formed by the cooperation of the three amino-terminal domains of one subunit with the four carboxyl-terminal domains of the other subunit. PMID- 8226760 TI - PA28, an activator of the 20 S proteasome, is inactivated by proteolytic modification at its carboxyl terminus. AB - PA28, a protein activator of the 20 S proteasome, was previously identified in soluble extracts of bovine red blood cells (Ma, C.-P., Slaughter, C. A., and DeMartino, G. N. (1992) J. Biol. Chem. 267, 10515-10523). To determine whether this regulatory protein is as widely distributed as the proteasome, PA28 content and activity were examined in various eukaryotic tissues by immunoblot analysis and by functional assays of tissue extracts. PA28 protein was present in all sources examined. PA28 activity, however, was not detected in many of these sources, including those with the highest level of PA28 protein. To determine the biochemical basis of this result, PA28 was purified from extracts of rat liver, which had high levels of PA28 protein but no PA28 activity. The resulting purified PA28 had no detectable activity but had native and subunit molecular weights indistinguishable from the active PA28 of bovine red blood cells. Using the inactivation of purified PA28 as an assay, a protein that inactivated PA28 without altering its apparent molecular weight on SDS-polyacrylamide gel electrophoresis was identified, purified, and characterized from bovine liver. It had biochemical and catalytic characteristics similar to those of lysosomal carboxypeptidase B. When leupeptin, an inhibitor of lysosomal carboxypeptidase B, was included in the buffers used for the preparation of PA28, PA28 activity was detected in tissues which otherwise failed to demonstrate this activity. A similar result was obtained when extracts were prepared in a manner that minimized disruption of lysosomes. Other carboxypeptidases such as carboxypeptidase Y and pancreatic carboxypeptidase B also inactivated PA28 without altering its apparent molecular weight. Active PA28 binds to the proteasome to form a protease-activator complex that can be isolated after velocity sedimentation centrifugation through glycerol density gradients. Carboxypeptidase-inactivated PA28 failed to form such a complex, suggesting that the carboxyl terminus of PA28 is required for binding to the proteasome. These results indicate the importance of the carboxyl terminus of PA28 for proteasome activation. PMID- 8226761 TI - Mitogenic and receptor activities of human growth hormone 108-129. AB - We have selectively synthesized a number of peptides encompassing the region of helix 3 of growth hormone (GH). These peptides and native human (h) GH have been evaluated for mitogenic and receptor activities in 3T3-F442A preadipocytes. In this system, wild type hGH is anti-mitogenic. In contrast, hGH 108-129 stimulated DNA synthesis while other GH-derived peptides were ineffective. hGH (L) 108-129 had an EC50 of about 0.2 nM and was maximally effective at about 0.5 nM in stimulating [3H]thymidine incorporation in 3T3-F442A cells. hGH (L) 108-129 was mitogenically as active as insulin-like growth factor-I and more active than insulin. It was less effective than transforming growth factor-beta. By cell cycle analysis, hGH (L) 108-129 increased the proportion of cells in S/G2/M phases to 28%. hGH, when coincubated with hGH (L) 108-129, blocked the mitogenic response of the peptide. A monoclonal antibody to the GH receptor significantly reduced binding of 125I-hGH to its receptor but had no effect on binding of 125I hGH (L) 108-129. Affinity cross-linking of 125I-hGH to its receptor was not duplicated with 125I-hGH (L) 108-129. No other GH peptides or insulin competed for binding of 125I-hGH 108-129. Scatchard analysis indicated a Kd of 5.2 nM with 5.6 x 10(5) binding sites/cell for hGH (L) 108-129. These studies indicate that hGH (L) 108-129, a sequence encompassing helix 3 of hGH, acts by binding to a site other than the GH receptor and evokes high mitogenic responses. PMID- 8226762 TI - Analysis of two distinct single-stranded DNA binding sites on the recA nucleoprotein filament. AB - The binding stoichiometry of Escherichia coli recA protein to single-stranded DNA (ssDNA) determined by two separate assays differs by a factor of 2.2-2.4. Using the fluorescence of etheno-DNA (epsilon DNA), a chemically modified ssDNA, the stoichiometry was found to be 7.0 +/- 0.6 bases/recA protein monomer in a nucleo protein filament. Using a competition assay, a similar stoichiometry, 7.5 bases/recA, is found for unmodified poly(dT). Using the DNA-dependent ATPase of recA, which monitors bound protein rather than bound DNA, we find that each recA monomer needs to bind only 3.1 +/- 0.5 bases to fully activate the ATPase. The difference in site size determined by the two assays indicates that there are two DNA binding sites with differential effects on ATPase activation. When recA protein is mixed with ssDNA at a ratio of 7 bases/recA or greater, the complex that forms contains 7 bases/recA and acts as a kinetic trap for the ssDNA. Upon further addition of recA protein, no additional ATPase activity is observed. If, on the other hand, the ssDNA is initially mixed with excess recA (at a ratio of 3 3.5 bases/recA or less) the ATPase activity is twice as high. Analysis of the binding curves suggests that the first DNA strand binds recA to form a filament with a stoichiometry of 3-3.5 bases/protein monomer. The ATPase activity of recA is completely active in this complex. A second strand of DNA can then be bound to this filament yielding a final stoichiometry of approximately 7 bases/protein monomer. The presence of this second strand neither enhances nor inhibits ATP hydrolysis. This ternary complex may mimic the structures formed by recA in searching for homologous DNA sequences and/or in the strand exchange reaction. PMID- 8226763 TI - Effects of Ca2+ binding on the protease module of factor Xa and its interaction with factor Va. Evidence for two Gla-independent Ca(2+)-binding sites in factor Xa. AB - The assembly of macromolecular complexes containing factors Xa and Va on suitable phospholipid surfaces is crucial for rapid activation of prothrombin. We have used quantitative affinity chromatography to characterize the interaction between factor Va and intact factor Xa on the one hand and between factor Va and factor Xa lacking the gamma-carboxyglutamic acid (Gla)-containing module on the other. The dissociation constants were found to be 1.0 +/- 0.1 and 9.5 +/- 1.8 microM, respectively. There was good agreement between these dissociation constants and the concentrations of active site-inhibited factor Xa and Gla-domainless factor Xa that caused half-maximal inhibition of prothrombin activation. To investigate whether the noncatalytic modules of factor Xa interacted directly with factor Va, intact modules were isolated from proteolytic digests of factor X and used as inhibitors of prothrombin activation. The inhibitory effect observed with the isolated Gla module in the absence of phospholipid was due to inhibition of the amidolytic activity of factor Xa rather than to an interaction with factor Va. The epidermal growth factor-like modules did not inhibit prothrombin activation. Using antibodies specific for calcium-dependent epitopes in the serine protease module of factor Xa we demonstrated that Ca2+ binding to the Gla module alters the conformation of the catalytic module. Half-maximal binding was observed at approximately 0.8 mM Ca2+. Evidence was also obtained for the presence of two Gla independent Ca(2+)-binding sites in factor Xa. One of these sites, located in the NH2-terminal epidermal growth factor-like module, was half-saturated at approximately 60 microM Ca2+ in intact factor Xa and at approximately 1.2 mM Ca2+ in Gla-domainless factor Xa. This site appeared not to influence the conformation of the protease module. The second site, which was half-saturated at approximately 0.16 mM Ca2+, appeared to reside in the serine protease module and to alter its conformation as judged by binding of antibodies specific for calcium dependent epitopes. PMID- 8226764 TI - Regulation of hepatic 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase gene expression by glucagon. AB - The control of hepatic 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase gene expression by glucagon was studied. Intraperitoneal administration of glucagon rapidly decreased the fructose 2,6-bisphosphate content by phosphorylation of 6 phosphofructo-2-kinase/fructose 2,6-bisphosphatase and diminution of its Vmax. Immunologic studies using a specific liver antibody showed that the amount of enzyme rapidly decreased. Northern blot analysis showed that the isozyme expressed is the adult liver form. The 6-phosphofructo-2-kinase/fructose 2,6 bisphosphatase mRNA content decreased, whereas that of phosphoenolpyruvate carboxykinase increased, and that of albumin did not change. Run-on transcription assays with isolated nuclei showed inhibition in the relative transcription rate of the 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase gene and a stimulation of phosphoenolpyruvate carboxykinase gene. The regulation of mRNA stability of 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase by glucagon was also studied. The half-life of mRNA decreased in the presence of glucagon, suggesting that proteins modulated by a glucagon-dependent process are regulating its stability. The time course of mRNA levels correlated with the transcription inhibition of gene and destabilization of mRNA, indicating that glucagon modulates 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase gene expression at both transcriptional and posttranscriptional levels. PMID- 8226765 TI - Identification of the functional domains of the FLP recombinase. Separation of the nonspecific and specific DNA-binding, cleavage, and ligation domains. AB - The FLP recombinase of the 2-microns plasmid of Saccharomyces cerevisiae binds to its recognition target (FRT) site, induces a bend in the DNA, and promotes DNA cleavage and strand ligation. We have subjected this protein to limited proteolysis and have purified three polypeptides: P13 (13 kDa), P21 (21 kDa), and P32 (32 kDa). These peptides are derived from the following regions of FLP: P13, amino acids 2-123; P21, amino acids 148-346; P32, amino acids 124-423. In this report, we show that P13 binds to DNA nonspecifically and P32, like P21, binds to the FRT site specifically. A single molecule of P32 is able to induce a bend in the DNA of 55 degrees, similar to that induced by intact FLP (63 degrees). P13 enhances the binding of P21 or P32 to the FRT site. Both P32 and P21 can catalyze DNA ligation in combination with P13. P32 can cleave and covalently attach to the FRT site in combination with P13, whereas P21 cannot. These results suggest that FLP contains two DNA-binding domains. A nonspecific DNA-binding region is located in the NH2-terminal 123 amino acids, whereas the region that imparts specific DNA binding resides in amino acids 148-346. Only the regions in P13 and P21 are needed for ligation activity, but those in P13 and P32 are required for cleavage. PMID- 8226766 TI - Binding of AP-1 Golgi adaptors to membranes requires phosphorylated cytoplasmic domains of the mannose 6-phosphate/insulin-like growth factor II receptor. AB - In mammalian cells, clathrin-coated vesicles mediate transport of the lysosomal enzyme receptors from the trans-Golgi network to the endocytic pathway. A critical step of this process is the recruitment of Golgi-specific adaptors onto Golgi membranes for efficient clathrin polymerization. An in vitro assay was used here to quantitate this event in streptolysin-O-permeabilized NRK cells. At 37 degrees C, these interactions are cytosol- and energy-dependent, sensitive to GTP gamma S (guanosine 5'-O-(thiotriphosphate)) and brefeldin A. We report that Golgi specific adaptor binding is enhanced in mannose 6-phosphate/insulin-like growth factor II (IGF II) receptor-overexpressing cells and reduced in mannose 6 phosphate receptor-deficient cells. Furthermore, adaptor binding is partially inhibited after addition of soluble cytoplasmic domains of the mannose 6 phosphate/IGF II receptor. Almost complete inhibition is only observed when this domain is phosphorylated on serines 2421 and 2492, a major modification acquired during exit of the receptor from the Golgi. These results show that the mannose 6 phosphate/IGF II receptor is part of the components that recruit the Golgi specific adaptors and that its phosphorylation is an important feature for high affinity interactions with sorting components. PMID- 8226767 TI - SH2 domains of the protein-tyrosine kinases Blk, Lyn, and Fyn(T) bind distinct sets of phosphoproteins from B lymphocytes. AB - Several members of the Src family, including Blk, Lyn, Fyn(T), and Lck, are expressed in B cells. These kinases associate with the antigen receptor complex, and the activities of Blk, Fyn(T), and Lyn increase upon receptor engagement. Differences in the amino acid sequences and patterns of expression of these kinases suggest that they serve distinct functions. In this communication it is shown that the SH2 domains from Blk, Lyn, and Fyn(T) preferentially bind distinct sets of phosphoproteins from the mature B cell line A20. These interactions were found to depend on recognition of phosphotyrosine. The Blk SH2 domain bound more than 10 distinct phosphoprotein species, most of which reacted with an antiphosphotyrosine antibody; the phosphotyrosine content of these proteins was increased if surface immunoglobulin was cross-linked before extracts were made. Phosphoproteins of 72, 76, 115, and 130 kDa bound to the SH2 domains of Blk, Lyn, and Fyn(T). Phosphoamino acid analysis of these four proteins revealed that each contained phosphoserine, phosphothreonine, and phosphotyrosine. Proteins of 90 kDa, 130 kDa, and 150 kDa were preferentially bound by the Blk SH2 domain, while the Fyn(T) SH2 domain showed preferential binding to proteins of 76 and 180 kDa. The Lyn SH2 binding profile resembled that of Blk, but differences in the binding specificities of these kinases were also observed. Thus, among proteins that exhibit increased tyrosine phosphorylation following antigen receptor cross linking, several have been identified that bind preferentially to SH2 domains of Blk, Fyn(T), or Lyn, suggesting that these kinases serve distinct functions. In addition, chimeric Fyn(T)-Blk SH2 domains were shown to be functional in binding assays and to exhibit binding specificities intermediate between those of the parent domains, consistent with the interpretation that the differences we observe in phosphoprotein binding by Fyn(T) and Blk SH2 domains reflect differences in their native structures. PMID- 8226768 TI - Poly(ADP-ribose) polymerase is a catalytic dimer and the automodification reaction is intermolecular. AB - We have determined the molecular mechanism of the automodification reaction of poly(ADP-ribose) polymerase (PARP) (EC 2.4.2.30). While PARP-mono(ADP-ribose) conjugates were the predominant products of automodification at 200 nM NAD, enzyme-bound branched polymers were preferentially synthesized at 200 microM NAD. Thus, the initiation, elongation, and branching reactions catalyzed by PARP appear to be [NAD]-dependent. Initial rates of automodification increased with second order kinetics as a function of [PARP] at both 200 nM and 200 microM NAD. Therefore, 2 molecules of PARP, i.e. a catalytic dimer, are required for the auto mono(ADP-ribosyl)ation and the auto-poly(ADP-ribosyl)ation reactions. Initial rates of automodification also increased with second order kinetics at low NAD concentrations. Therefore, the catalytic dimer also requires 2 molecules of NAD. These results are consistent with the conclusion that the automodification reaction of PARP is intermolecular and that the 2 monomeric units of PARP may simultaneously function as catalyst and acceptor molecules in the automodification reaction. Confirmatory evidence for the catalytic role of protein-protein interactions in the automodification reaction was manifested by a marked inhibition of auto-poly(ADP-ribosyl)ation at 40 nM or higher [PARP]. PMID- 8226769 TI - Isolation and characterization of ClpX, a new ATP-dependent specificity component of the Clp protease of Escherichia coli. AB - We have used 14C-labeled bacteriophage lambda O-DNA replication protein as a probe to identify and purify Escherichia coli proteases capable of its degradation. In this manner, five different proteases (termed Lop) have been identified capable of degrading lambda O protein to acid-soluble fragments in an ATP-dependent fashion. One of these activities was purified to homogeneity and shown to be composed of two different polypeptides. The 23,000-Da component (LopP) was identified as the previously characterized ClpP protein, known to complex with ClpA to form the ClpAP, an ATP-dependent protease, capable of degrading casein. The second 46,000-Da component was identified as ClpX (LopC), coded by a gene located in the same operon, but promoter distal to that coding for ClpP (Gottesman, S., Clark, W. P., de Crecy-Lagard, V., and Maurizi, M. R. (1993) J. Biol. Chem. 268, 22618-22626). This identification was based on the determination of the sequence of the first 24 amino acid residues of the purified ClpX protein and its identity with that predicted by the DNA sequence. The ClpXP protease is substrate specific, since it degrades casein (known to be degraded by ClpAP), lambda P, or DnaK proteins slowly or not at all. These results suggest that ClpX protein directs ClpP protease to specific substrates. It is estimated that 50% of all lambda O-specific protease activity present in crude E. coli extracts is due to the ClpXP protease. We propose that transient inhibition of lambda O degradation observed in vivo during the later stages of lambda-DNA replication in vivo is responsible for the switch from bidirectional to unidirectional replication. One round unidirectional replication will lead to strand separation resulting in a switch from early (theta) to late (sigma) mode of lambda-DNA replication. PMID- 8226770 TI - ClpX, an alternative subunit for the ATP-dependent Clp protease of Escherichia coli. Sequence and in vivo activities. AB - The ATP-dependent Clp protease of Escherichia coli consists of two subunits, the ClpP subunit, which has the proteolytic active site, and ClpA, which possesses ATPase activity and activates the proteolytic activity of ClpP in vitro. Recently, Zylicz and co-workers (Wojtkowiak, D., Georgopoulos, C., and Zylicz, M. (1993) J. Biol. Chem. 268, 22609-22617) identified another E. coli protein that activated ATP-dependent degradation of lambda O protein in the presence of ClpP. The amino-terminal sequence of this protein corresponds to the translated amino terminal sequence of a gene that we have named clpX. clpX encodes a protein with M(r) 46,300, similar to that observed for the protein purified by Wojtkowiak et al. clpX is an operon with clpP; both genes are cotranscribed in a single heat inducible 2200-base mRNA, with clpP the promoter proximal gene. The sequence of ClpX includes a single consensus ATP-binding site motif and has limited homology to regions of ClpA and other members of the ClpA/B/C family. A third group of proteins, ClpY, closely related to ClpX, has been identified by sequence homology. Mutations in either clpX or clpP abolish degradation of the highly unstable lambda O protein in vivo. clpX mutants are not defective in degradation of previously identified ClpA/ClpP substrates such as a ClpA-beta-galactosidase fusion protein. It appears that selectivity of degradation by ClpP in vivo is determined by interaction of ClpP with different regulatory ATPase subunits. PMID- 8226771 TI - C-terminal processing of human beta-glucuronidase. The propeptide is required for full expression of catalytic activity, intracellular retention, and proper phosphorylation. AB - beta-Glucuronidase undergoes proteolytic C-terminal processing during or after its transport to lysosomes or endosomes. We determined the C-terminal processing site for human placental beta-glucuronidase to be the peptide bond between Thr633 Arg634. To evaluate the role of the 18-amino acid peptide removed in C-terminal processing, we changed the codon for Arg634 to a stop codon by site-directed mutagenesis and studied expression of the truncated mutant enzyme in COS-7 cells. An increased fraction of newly synthesized enzyme from R634Stop cDNA was secreted. Pulse-chase experiments provided no evidence for increased degradation of the intracellular R634Stop enzyme. The total amount of catalytic activity expressed from the R634Stop mutant cDNA was only half that seen with the wild type cDNA, and the Kcat of the mutant enzyme was 52% that of wild type enzyme. These results indicate that the C-terminal propeptide in the precursor is important for beta-glucuronidase to achieve maximal activity. The truncated enzyme formed hybrid tetramers in cotransfection experiments with the cDNA for rat beta-glucuronidase. There appeared to be no decrease in stability of the R634Stop enzyme, since chaotropic agents, heat treatment, and pH had similar effects on the mutant and the wild type enzymes. The uptake rate of the truncated mutant (R634Stop) enzyme by beta-glucuronidase-deficient human fibroblast cells was only 55-60% that of the wild type enzyme. Binding to the immobilized cation independent mannose-6-phosphate receptor and measurement of the 32P-labeled phosphorylated oligosaccharides revealed that the truncated mutant enzyme was 32 34% less phosphorylated and appeared to contain proportionately more covered phosphate groups than the wild type enzyme. These results suggest that the propeptide influences the accessibility to both processing enzymes that produce the mannose-6-phosphate recognition marker on beta-glucuronidase. PMID- 8226772 TI - Positioning of major tryptic fragments in the Ca2+ release channel (ryanodine receptor) resulting from partial digestion of rabbit skeletal muscle sarcoplasmic reticulum. AB - Site-specific antibodies against different regions of the Ca2+ release channel of skeletal muscle sarcoplasmic reticulum (ryanodine receptor) were developed and used as probes for immunoblotting of the major tryptic fragments resulting from partial digestion of the ryanodine receptor in sarcoplasmic reticulum membranes. Five major tryptic fragments, some of which migrated as doublets, with apparent masses of 150/140, 110/100, 55, 170/160, and 76 kDa were ordered so that they covered the bulk of the protein from the NH2 to the COOH terminus. Tryptic subfragments of 53, 63, and 115/95 kDa were also derived from the 150/140-, 110/100-, and 170/160-kDa fragments, respectively. All of these fragments and subfragments were detected only in the insoluble membrane fraction of the trypsinized sarcoplasmic reticulum. Upon Na2CO3 extraction, the 150/140-, 110/100 , and 55-kDa fragments could be solubilized, suggesting their origin in the cytoplasmic domain of the ryanodine receptor. The 170/160- and 76-kDa fragments and the 115/95-kDa subfragment remained insoluble, suggesting their origin in the transmembrane region of the ryanodine receptor. The 150/140-, 110/100-, 170/160-, and 76-kDa fragments and the 115/95 subfragment co-migrated near the bottom of a sucrose density gradient after CHAPS solubilization, suggesting that they were associated in an oligomeric complex. By contrast, the 53- and 63-kDa subfragments and the 55-kDa fragment were detected near the top of the sucrose gradient after CHAPS solubilization, suggesting that they were not involved in the formation of the core of the oligomeric complex. These studies identify 7 sites that are exposed to trypsin in the ryanodine receptor in sarcoplasmic reticulum, 3 of which are novel and 4 of which are in the same location as proteolytic cleavage sites identified previously (Marks, A. R., Fleischer, S., and Tempst, P. (1990) J. Biol. Chem. 265, 13143-13149). PMID- 8226773 TI - Functional characterization of promoter elements involved in regulation of human B beta-fibrinogen expression. Evidence for binding of novel activator and repressor proteins. AB - A high level of plasma fibrinogen has been shown to be an important risk factor for myocardial infarction and stroke. Thus, we were prompted to investigate regulation of human fibrinogen biosynthesis, a process wherein expression of the B beta-chain of fibrinogen appears to be rate-limiting for fibrinogen secretion. Using electrophoretic mobility shift assays with synthetic probes representing portions of the human B beta-fibrinogen promoter, we have defined several elements that bind distinct classes of transcription factors present in human hepatoma cell nuclear extracts. The contribution of each element to promoter activity was demonstrated in transfection experiments using promoter chloramphenicol acetyltransferase constructs and human hepatoma cells. Our observations indicate that two distinct sequence elements are required for maximal induction of transcription by interleukin-6. One of these sequences is an IL-6-RE core element similar to that reported for the rat alpha 2-macroglobulin promoter and the other is a binding site for the C/EBP family of transcription factors. We also report two additional elements, one negative- and one positive acting, that bind novel sequence-specific factors. PMID- 8226774 TI - Spectrin cagliari. an Ala-->Gly substitution in helix 1 of beta spectrin repeat 17 that severely disrupts the structure and self-association of the erythrocyte spectrin heterodimer. AB - The spectrin tetramer, the principal structural element of the red cell membrane skeleton, is formed by stable head-to-head self-association of two spectrin heterodimers. The self-association site appears to be formed by interactions between helices 1 and 2 of beta spectrin repeat 17 of one dimer with helix 3 of alpha spectrin repeat 1 of the other dimer to form two combined alpha-beta triple helical segments. The head of the heterodimer appears to involve similar intradimer interactions. We describe the first example of an amino acid substitution in helix 1 of this combined alpha-beta triple-helical segment, which, although relatively minor, profoundly impairs tetramer formation. Strikingly, low angle rotary shadowing electron microscopy of isolated spectrin dimers reveals that this mutation also severely disrupts the head of the heterodimer causing it to be open. Following linkage studies which were most consistent with a beta spectrin gene mutation, a nucleotide change was identified in codon 2018, resulting in an Ala-->Gly substitution in the first helical domain of beta spectrin repeat 17. Because glycine is a strong helix breaker, this change is predicted to disrupt the conformation of this helical domain. Our results indicate that this helical domain must play direct roles in the alpha beta interdimer interactions that form the self-association site of the tetramer and in the alpha-beta intradimer interactions at the head of the heterodimer. PMID- 8226775 TI - Protein chemical characterization and immunocytochemical localization of the NMDA receptor subunit NMDA R1. AB - In the rat central nervous system, the mRNA encoding the N-methyl-D-aspartate receptor subunit R1 is the most ubiquitously distributed among the cloned subunit mRNAs of this glutamate receptor subtype. The N-methyl-D-aspartate R1 mRNA is very abundantly expressed and N-methyl-D-aspartate R1 coexpression is necessary for functional expression of all other cloned N-methyl-D-aspartate receptor subunits. Therefore, the R1 subunit is likely to be an essential component of all known N-methyl-D-aspartate receptors in rat brain. By employing sequence specific polyclonal antibodies, we demonstrate that rat brain N-methyl-D-aspartate R1, as well as recombinantly expressed receptor protein, has an apparent molecular mass of 116 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis. The receptor protein is heavily glycosylated. It is specifically localized to the central nervous system, and it co-enriches with synaptic membranes upon subcellular fractionation of the cerebral cortex. Chemical cross-linking of synaptic membrane proteins shows that the N-methyl-D-aspartate R1 protein is part of a receptor protein complex with a molecular mass of 730 kDa. By using immunocytochemical methods, we demonstrate a widespread but distinct distribution of N-methyl-D-aspartate R1 in neurons of the rat brain, with prominent immunostaining in certain layers of the cerebral cortex, in the hippocampus and dentate gyrus, as well as in the cerebellum. PMID- 8226776 TI - Artificial duplication of the R67 dihydrofolate reductase gene to create protein asymmetry. Effects on protein activity and folding. AB - R67 dihydrofolate reductase (DHFR), encoded by an R plasmid, provides resistance to the antibacterial drug trimethoprim. This enzyme does not exhibit any structural or sequence homologies with chromosomal DHFR. A recent crystal structure of tetrameric R67 DHFR (D. Matthews, X. Nguyen-huu, and N. Narayana, personal communication) shows a single pore traversing the length of the molecule. Numerous physical and kinetic experiments suggest the pore is the active site. Since the center of the pore possesses exact 222 symmetry, mutagenesis of residues designed to explore substrate binding will probably also affect cofactor binding. As a first step in breaking this inevitable symmetry in R67 DHFR, the gene has been duplicated. The protein product, R67 DHFRdouble, is twice the molecular mass of native R67 DHFR and is fully active with kcat = 1.2 s 1, Km(NADPH) = 2.7 microM and Km(dihydrofolate) = 6.3 microM. Equilibrium unfolding studies in guanidine-HCl indicate R67 DHFRdouble is more stable than native R67 DHFR at physically reasonable protein concentrations. Microcalorimetry studies show native R67 DHFR undergoes fully reversible thermal unfolding. Unfolding can be described by a two-state process since a ratio of delta Hcalorimetric to delta Hvan't Hoff equals 0.96. In contrast, thermal unfolding of R67 DHFRdouble is not fully reversible and possesses an oligomerization component introduced by the gene duplication event. PMID- 8226777 TI - Cloning and functional expression of a Dictyostelium discoideum protein tyrosine phosphatase. AB - Using polymerase chain reaction methods, we cloned a 1.7-kilobase cDNA, denoted DdPTPa, that has high homology with other known eukaryotic protein tyrosine phosphatases. DdPTPa possess a 241-amino acid protein tyrosine phosphatase domain located in the C terminus, which exhibits a 39-43% amino acid sequence identity with published protein tyrosine phosphatases. Absence of a characteristic signal sequence and transmembrane domain suggests that DdPTPa is a nonreceptor type cytoplasmic protein tyrosine phosphatase. Southern blot analysis of genomic DNA indicates the presence of a multigene protein tyrosine phosphatase family in Dictyostelium. Northern blot analysis reveals four species of mRNA that hybridize to the DdPTPa probe, at least three of which are developmentally regulated. The entire coding sequence of DdPTPa was subcloned into the pET15-b vector and expressed in Escherichia coli. Affinity-purified DdPTPa protein efficiently dephosphorylates both p-nitrophenyl phosphate and tyrosine-phosphorylated reduced, carboxyamidomethylated, and maleylated lysozyme. A Dictyostelium transformant overexpressing DdPTPa does not develop normally. The overexpresser fails to aggregate, in contrast to the control transformant containing vector alone, and after 24 h gives rise to only a few abnormal slugs and small fruiting bodies. PMID- 8226778 TI - Kinetic analysis of ouabain binding to native and mutated forms of Na,K-ATPase and identification of a new region involved in cardiac glycoside interactions. AB - Cardiac glycosides inhibit the Na,K-ATPase by binding to the catalytic alpha subunit of the enzyme. Site-directed mutagenesis of the H1-H2 domain has demonstrated the importance of this region in determining cardiac glycoside affinity. In this study, random mutagenesis was used to identify an amino acid, arginine 880, in the COOH-terminal portion of the alpha subunit which influences the sensitivity of the enzyme to ouabain. This residue is predicted to reside in the H7-H8 extracellular loop. Conversion of arginine 880 to a proline causes a 10 fold increase in the dissociation rate constant and a 2-fold increase in the association rate constant for [3H]ouabain binding. This results in an enzyme with a KD for ouabain 5-fold higher than the wild-type sheep alpha 1 isoform. These data are compatible with arginine 880 comprising a portion of the ouabain binding site. Furthermore, if arginine 880 is at the physical binding site, then this finding lends support to models that place this amino acid extracellularly since cardiac glycosides interact with the extracellular surface of the Na,K-ATPase. The ouabain binding characteristics of substitution R880P were compared with those of several different Na,K-ATPases, each of which contains a single amino acid substitution in the H1-H2 region of the alpha subunit. The substituted enzymes, C104A, Y108A, E116Q, P118K, and Y124F, vary considerably in their rates of dissociation (1-4-fold increase in the dissociation rate constant). In addition, the rate of association of [3H]ouabain binding to substitution P118K is 2-fold slower than that of the wild-type enzyme. These results suggest that the H1-H2 domain may participate directly in ouabain binding as well as be involved in conformational changes, both of which could affect the sensitivity of the enzyme to ouabain. PMID- 8226779 TI - Interaction of the tail domain of high molecular weight subunits of neurofilaments with the COOH-terminal region of tubulin and its regulation by tau protein kinase II. AB - We previously showed that neurofilaments interact with microtubules (MTs) via their high molecular weight subunits (NF-H) after alkaline phosphatase treatment. Here we studied the effects of phosphorylation of NF-H on this interaction. tau protein kinase II, Ser/Thr protein kinase, phosphorylated NF-H in the tail domain, decreased its electrophoretic mobility to a native level, and also restored its property to be less interactive with MTs. Phosphorylation by cAMP dependent protein kinase caused no shift of electrophoretic mobility or dissociation from MTs. We conclude that the tail domain of NF-H directly interacts with the MT surface, and the interaction is regulated via phosphorylation of the tail domain of NF-H by Ser/Thr protein kinase like tau protein kinase II. To characterize the binding domain of NF-H on MTs, subtilisin digestion of MTs and competition analysis with the MT binding fragment of tau protein were performed. The dissociation constant of NF-H to subtilisin MTs was higher than that to intact MTs. The maximum binding of NF-H was reduced when tau fragments existed. These results revealed that the COOH-terminal region of tubulin is involved in the binding to NF-H, and the NF-H and microtubule associated protein binding domains are closely apposed on the surface of MTs. PMID- 8226780 TI - The membrane-binding domain of ankyrin contains four independently folded subdomains, each comprised of six ankyrin repeats. AB - Ankyrin repeats are a 33-amino acid motif present in a number of proteins of diverse functions including transcription factors, cell differentiation molecules, and structural proteins. This motif has been shown to mediate protein interactions in the case of ankyrin as well as several other repeat-bearing proteins. In ankyrin, 24 tandemly arrayed repeats are arranged to form a globular, membrane-binding domain. This report provides evidence that the repeats in this domain fold into four independently folded subdomains of six repeats each. Limited proteolytic digestions of defined regions of the membrane-binding domain identified protease-sensitive sites, which divided this domain into subdomains of approximately six repeats each. Hydrodynamic measurements and circular dichroism spectroscopy of expressed subdomains confirmed that these six repeat regions exist as folded, globular structures. The requirement of a complete set of six repeats for proper folding was determined using a series of protein constructs, which sequentially deleted repeats from the last subdomain. Deletion of even one repeat resulted in a 40% loss of alpha-helicity. Deletions removing three or more repeats abolished the helical signal completely. The spherical shapes of the intact domain and of the subdomains (inferred from hydrodynamic values) suggest that the four subdomains are organized in either a tetrahedral or square planar configuration. Two six-repeat subdomains were found to be required for high affinity association with the anion exchanger, suggesting that at least some of the protein interactions mediated by ankyrin repeats involve multiple subdomains. PMID- 8226781 TI - Localization of transforming growth factor beta binding site in betaglycan. Comparison with small extracellular matrix proteoglycans. AB - The most abundant binding molecule for transforming growth factor beta (TGF-beta) on many cell types is betaglycan, a transmembrane proteoglycan. To localize the binding site for TGF-beta in betaglycan, parts of the betaglycan extracellular portion were expressed as recombinant fusion proteins in bacteria and tested for their ability to compete for the binding of TGF-beta 1 to Hep G2 cells. One fragment encompassing 226 residues near the transmembrane domain (amino acids 543 769) (Lopez-Casillas, F., Cheifetz, S., Doody, J., Andres, J. L., Lane, W. S., and Massague, J. (1991) Cell 67, 785-795) was active, whereas fusion proteins representing the other parts of the betaglycan ectodomain were inactive. Affinity measurements revealed two classes of binding sites (Kd = 3.9 nM and Kd = 145 nM) for the active fusion protein. The binding of the betaglycan fusion protein to immobilized TGF-beta 1 was inhibited by fusion proteins representing the core proteins of the small interstitial proteoglycans decorin, biglycan and fibromodulin, each also known to bind TGF-beta. The effective concentrations of TGF-beta for binding to these other proteoglycans were similar to those required for binding to betaglycan, indicating similar affinities for the binding of proteoglycans and betaglycan. Affinity cross-linking showed that, at low concentrations, the betaglycan fragment enhanced the binding of TGF-beta to the type II receptor and to endogenous betaglycan but had no effect on the binding to type I receptor. At high concentrations, the TGF-beta binding fragment inhibited the binding of TGF-beta to all these receptors. The fragment enhanced the activity of TGF-beta in mink lung cell bioassay at all active concentrations. The results indicate that betaglycan and the decorin type proteoglycans all bind to the same or closely spaced sites in TGF-beta and compete with one another for the binding. In addition, betaglycan may cooperate with the type II receptor on TGF beta binding. PMID- 8226782 TI - The human cot proto-oncogene encodes two protein serine/threonine kinases with different transforming activities by alternative initiation of translation. AB - The cot gene is an oncogene encoding serine/threonine kinases isolated by DNA transfection assay. In this study, we isolated cDNA for the human cot protooncogene (proto-cot gene) and examined the structure and function of its gene products. The proto-cot gene has an open reading frame encoding 467 amino acids of which the first 397 amino acids are identical to those in the corresponding part of the cot gene. The protein products of the proto-cot gene were identified as 58- and 52-kDa proteins with intrinsic protein serine/threonine kinase activity. These two protein species were suggested to be generated by alternative initiation from two AUGs. The 58- and 52-kDa proteins are both localized predominantly in the cytosol, but the 58-kDa protein has a shorter half-life than the 52-kDa protein, suggesting the importance of the amino terminal domain in regulating the stability of the proto-Cot protein. More interestingly, the 58-kDa protein showed stronger transforming activity than the 52-kDa protein, although this activity was much weaker than that of the Cot oncoprotein. Thus, the amino-terminal domain of the Cot protein may be necessary for cellular transformation, whereas the carboxyl-terminal domain may negatively regulate the transforming activity. PMID- 8226783 TI - Tyrosine kinase(s) regulate apoptosis and bcl-2 expression in a growth factor dependent cell line. AB - Apoptosis (programmed cell death) plays a critical role in many physiological processes, but the mechanism(s) which regulate apoptosis are poorly understood. We demonstrate that in a hematopoietic cell line, which can grow in either interleukin (IL)-2 or IL-3, both of these growth factors can increase bcl-2 mRNA levels and prevent apoptosis normally seen following growth factor withdrawal. Herbimycin A, a protein tyrosine kinase inhibitor, blocks the ability of IL-2 and IL-3 to up-regulate bcl-2 mRNA levels and induces apoptosis. Transfection of a bcl-2 expression vector not only prolongs survival following growth factor withdrawal but also confers resistance to the effect of herbimycin A. We conclude that herbimycin A-sensitive protein tyrosine kinases are involved in the regulation of apoptosis and bcl-2 expression, but these protein tyrosine kinases appear not to be required for the action of Bcl-2 since Bcl-2 can exert its growth survival effect even in the presence of herbimycin A. PMID- 8226784 TI - Rapamycin inhibition of interleukin-2-dependent p33cdk2 and p34cdc2 kinase activation in T lymphocytes. AB - The immunosuppressant rapamycin (RAP) is a potent inhibitor of the entry of interleukin (IL)-2-stimulated T cells into S-phase. Earlier results indicated that RAP treatment arrested the growth of the murine IL-2-dependent T cell line CTLL-2 in late G1-phase. To explore further the interactions of RAP with the cell cycle control machinery in T cells, we examined the effects of RAP treatment on the activation of the cyclin-dependent kinases p34cdc2 and p33cdk2 in G1-phase CTLL-2 cells. Stimulation of factor-deprived cells with IL-2 led to the assembly of high molecular weight complexes containing active p34cdc2 and p33cdk2. The appearance of these complexes was explained, at least in part, by the association of both cyclin-dependent kinases with IL-2-induced cyclin A. RAP treatment profoundly inhibited both cyclin A expression and the appearance of active cyclin A-cyclin-dependent kinase complexes in IL-2-stimulated, late G1-phase CTLL-2 cells. Although p34cdc2 activation was largely dependent on association with cyclin A, a significant proportion of the active p33cdk2 pool was complexed with cyclin E. In contrast to cyclin A, the IL-2-induced accumulation of cyclin E in G1-phase cells was only partially suppressed by RAP, and cyclin E-p33cdk2 complexes were readily detected in drug-treated cells. These cyclin E-cyclin dependent kinase complexes were nonetheless devoid of histone H1 kinase activity. The inhibitory effects of RAP on the activation of cyclin E- and cyclin A associated cyclin-dependent kinases suggest that one or both events participate in the regulation of T cell entry into S-phase. PMID- 8226785 TI - Identification of residues in the single-stranded DNA-binding site of the 8-kDa domain of rat DNA polymerase beta by UV cross-linking. AB - Rat DNA polymerase beta (beta-pol) is a 39-kDa monomeric protein, organized in two structurally and functionally distinct domains. The 8-kDa NH2-terminal domain binds single-stranded (ss) DNA, whereas the 31-kDa COOH-terminal domain does not. To facilitate studies on ssDNA binding structure-function relationships of beta pol, we overexpressed the 8-kDa domain in Escherichia coli, and purified the recombinant protein to homogeneity. Single-stranded nucleic acid binding of the recombinant 8-kDa domain was found to be similar to that previously reported for the 8-kDa fragment prepared by proteolysis of intact beta-pol (Kumar, A., Widen, S. G., Williams, K. R., Kedar, P. Karpel, R. L., and Wilson, S. H. (1990b) J. Biol. Chem. 265, 2124-2131; Casas-Finet, J. R., Kumar, A., Morris, G., Wilson, S. H., and Karpel, R. L. (1991) J. Biol. Chem. 266, 19618-19625). Residues in or near the DNA-binding pocket of the recombinant 8-kDa domain were examined by photochemical cross-linking to [32P] p(dT)16. Cross-linking was localized to a tryptic fragment spanning residues 28 through 35 and a V8 protease fragment spanning residues 27 through 58. Sequence analysis of the various [32P]p(dT)16 labeled proteins indicated that Ser30 and His34 were modified by cross-linking to p(dT)16. Therefore, these residues of the ssDNA-binding domain of beta-pol appear to be in close contact with this nucleic acid probe. PMID- 8226786 TI - Localization and functional analysis of CHIP28k water channels in stably transfected Chinese hamster ovary cells. AB - CHIP28 is a major water transporting protein in erythrocytes and plasma membranes in kidney proximal tubule and thin descending limb of Henle. Chinese hamster ovary cells were stably transfected with the coding sequence of cloned rat kidney CHIP28k using expression vectors containing cytomegalovirus or Rous sarcoma virus promoters. Clonal cell populations expressed a 1.3-kilobase mRNA on Northern blot probed by CHIP28k cDNA and a 28-kDa protein on immunoblot probed by a polyclonal CHIP28 antibody. The clone with greatest expression produced approximately 8 x 10(6) copies of CHIP28k protein/cell. Plasma membrane osmotic water permeability (Pf), measured by stopped-flow light scattering, was 0.004 cm/s in control (vector-transfected) cells (10 degrees C) and 0.014 cm/s in the CHIP28k transfected cells. Pf in CHIP28k-transfected cells had an activation energy of 4.9 kcal/mol and was reversibly inhibited by HgCl2. CHIP28k expression did not affect the transport of protons and the small polar non-electrolytes urea and formamide. CHIP28k immunoreactivity and function was then determined in subcellular fractions. Pf in 6-carboxyfluorescein-labeled endocytic vesicles, measured by a stopped-flow fluorescence quenching assay, was 0.002 cm/s (control cells) and 0.011 cm/s (CHIP28k-transfected cells); Pf in transfected cells was inhibited by HgCl2. Immunoblotting of fractionated endoplasmic reticulum, Golgi, and plasma membranes revealed high densities of CHIP28k (approximately 5000 monomers/microns 2 in plasma membrane) with different glycosylation patterns; functional water transport activity was present only in Golgi and plasma membrane vesicles. Antibody detection of CHIP28k by confocal fluorescence microscopy and immunogold electron microscopy revealed localization to plasma membrane and intracellular vesicles. These studies establish a stably transfected somatic cell line that strongly expresses functional CHIP28k water channels. As in the original proximal tubule cells, the expressed CHIP28k protein is a selective water channel that is functional in endocytic vesicles and the cell plasma membrane. PMID- 8226787 TI - Expression, purification, and characterization of SH2-containing protein tyrosine phosphatase, SH-PTP2. AB - A human protein tyrosine phosphatase containing two src homology 2 (SH2) domains (SH-PTP2) was expressed in Escherichia coli under T7 promoter control and purified to near homogeneity. The purified protein, with molecular mass of 68 kDa on SDS-polyacrylamide gel electrophoresis, was identified as SH-PTP2 by its protein tyrosine phosphatase activity and N-terminal amino acid sequence analysis. Its protein tyrosine phosphatase activity was sensitive to pH and salt concentration. Whereas its optimum pH for the low molecular weight substrate para nitrophenyl phosphate is 5.6, the pH optima for peptide substrates were shifted toward neutral. With the artificial protein substrate reduced, carboxyamidomethylated, and maleylated lysozyme, it displays 2000-fold lower Km (1.7 microM) and 2.4-fold higher kcat (0.11 s-1) than with para-nitrophenyl phosphate. Among the phosphopeptides from autophosphorylation sites of receptors for epidermal growth factor and platelet-derived growth factor, SH-PTP2 displayed high activity toward phosphopeptides corresponding to pY992 of the epidermal growth factor receptor and pY1009 and pY1021 of the platelet-derived growth factor receptor. In further enzymatic studies with phosphopeptides corresponding to pY1009, SH-PTP2 showed nonlinear Line-weaver-Burk double-reciprocal plots, suggesting that the phosphopeptide corresponding to pY1009 may have a substrate and allosteric effect. PMID- 8226788 TI - Structure and regulation of the growth arrest-specific (gas-1) promoter. AB - We have isolated recombinant genomic clones encompassing several kilobase pairs of the 5'-flanking regions of both the human and murine gas-1 gene (growth arrest specific gene 1). Both species share a highly conserved region of approximately 550 base pairs upstream of the gas-1 transcription start site. Deletion analysis of the murine gas-1 promoter demonstrated that a fragment containing the first 665 base pairs is sufficient to drive the serum-regulated expression of a luciferase reporter gene in NIH3T3 cells, in a manner qualitatively reflecting the activity of the endogenous gene. Gel retardation assays indicated the presence of a number of DNA-binding proteins specific for sequences contained within the gas-1 transcription regulatory region. Comparative studies with extracts prepared from growing and resting cells revealed several growth state specific binding activities. One promoter fragment that bound prominent growth- and arrest-specific complexes was further analyzed by copper-phenanthroline footprinting. It was found that the same DNA element is a target both for growth- and for arrest-specific activities. The factors characterized in this study are the first candidates for transcriptional regulators mediating cell growth specific repression and/or growth arrest-specific activation of gene expression. PMID- 8226789 TI - Analysis of a pause transfer sequence from apolipoprotein B. AB - In contrast to typical secretory proteins, apolipoprotein B pauses at distinct points along the nascent chain during its translocation into the lumen of the endoplasmic reticulum. Specific pause transfer sequences mediate such discrete pauses in the translocation of apolipoprotein B. These sequences have been shown to confer this translocational behavior to heterologous chimeric proteins. To investigate the function of pause transfer sequences, we: (i) examine whether the multiple pause transfer sequences of apolipoprotein B act independently or are dependent upon the action of upstream pause transfer sequences, (ii) identify residues of the prototypical B' pause transfer sequence that are involved in pausing, and (iii) determine whether the stopping step of a translocational pause is a consequence of translational pausing, as has been suggested by other investigators. We conclude that pause transfer sequences act independently of each other and of translation; translocational pausing occurs even in the absence of ongoing protein synthesis. Furthermore, like other topogenic sequences such as signal sequences, pause transfer sequences are degenerate in structure yet have distinctive features necessary for their action. This characterization of the B' pause transfer sequence may aid in the identification of such sequences elsewhere in apolipoprotein B and in other proteins and has implications for the mechanism of translocational pausing. PMID- 8226790 TI - Cloning and expression of a spinal cord- and brain-specific glycine transporter with novel structural features. AB - A novel glycine transporter (GLYT2) was cloned from a rat brain cDNA library. GLYT2 is about 48 and 50% homologous to the previously cloned mouse glycine transporter (GLYT1) and rat proline transporter (PROT), respectively. GLYT2 differs from GLYT1 in molecular structure, tissue specificity, and pharmacological properties. The cDNA of GLYT2 encodes for 799 amino acid residues with an extended amino-terminal peptide containing 200 amino acids before the first transmembrane domain. Potential phosphorylation sites for protein kinase C, cAMP-dependent kinase, and calmodulin-dependent kinase were identified in the amino-terminal region. GLYT2 mRNA was shown to be specifically localized in spinal cord, brain stem, and to a lesser extent in the cerebellum. In contrast, GLYT1 mRNA distribution in the brain has been found previously to be more ubiquitous. Xenopus oocytes injected with GLYT2 cRNA transport glycine with a Km of 17 microM, and the uptake of glycine is resistant to inhibition by sarcosine. The experimental data suggests GLYT2 might play a major role in the termination of the inhibitory effect of glycine in the brain stem and spinal cord of vertebrates. On the other hand, the main function of GLYT1 may be in the modulation of excitatory nerve terminals. Two types of GLYT1 cDNA, GLYT1a and GLYT1b, were cloned from the mouse brain library. They differ only at their amino terminal sequences, and GLYT1b contains two additional potential phosphorylation sites for proline-dependent kinase. Cloning of the gene encoding the GLYT1 revealed that the two variants resulted from a differential splicing. PMID- 8226791 TI - Inhibition of internalization of glucose transporters and IGF-II receptors. Mechanism of action of MHC class I-derived peptides which augment the insulin response in rat adipose cells. AB - Peptides from the alpha 1 domain of the major histocompatibility complex class I antigen (MHC class I), e.g. Dk-(61-85) and Dk-(62-85), have been shown previously to augment glucose uptake in insulin-stimulated cells and to inhibit insulin receptor internalization (Stagsted, J., Reaven, G. M., Hansen, T., Goldstein, A., and Olsson, L. (1990) Cell 62, 297-307). We now report that these peptides inhibit by 80-100% the internalization of glucose transporters (GLUT4) and insulin-like growth factor II (IGF-II) receptors in insulin-stimulated cells and correspondingly double insulin-stimulated glucose transport activity and the number of GLUT4 and IGF-II receptors on the cell surface. In addition, the peptides enhance the apparent affinity about 3-fold of IGF-II binding to its receptor. It is concluded that the effects of the peptides on glucose transport and IGF-II binding are a consequence of the peptide-mediated inhibition of internalization of GLUT4 and IGF-II receptor. The active peptides are derived from the alpha 1 domain of a MHC class I molecule, suggesting that the latter is involved in regulation of internalization of cell surface integral membrane proteins such as the GLUT4 and IGF-II and insulin receptors. PMID- 8226792 TI - Dominant-negative mutations of platelet-derived growth factor (PDGF) receptors. Inhibition of receptor function by ligand-dependent formation of heterodimers between PDGF alpha- and beta-receptors. AB - We showed previously that a truncated form of the platelet-derived growth factor (PDGF) beta-receptor lacking its kinase region can form a nonfunctional heterodimer with the wild-type beta-receptor and thereby inhibit its signal transduction. In this paper we investigated whether the truncated form of either alpha- or beta-receptor could block the function of the other type of wild-type PDGF receptor. When the truncated alpha-receptor was expressed in Xenopus oocytes in excess over either the wild-type alpha- or beta-receptor, the Ca2+ mobilization signal elicited by either the wild-type alpha- or beta-receptor was completely blocked. The truncated beta-receptor abolished signaling by the wild type alpha-receptor in response to PDGF-AB or -BB. However signal transduction by the alpha-receptor in response to PDGF-AA was not affected by the truncated beta receptor. In the presence of PDGF-AB or -BB, both the wild-type and truncated beta-receptors formed a heterologous complex with the alpha-receptor in intact cells. A kinase-inactive beta-receptor (an ATP-binding site mutation) became cross-phosphorylated on tyrosine residues by the co-expressed wild-type alpha receptor in response to PDGF-AB or -BB but not in response to PDGF-AA. These findings indicate that the alpha-and beta-receptors interact in response to PDGF AB or -BB and are consistent with the hypothesis that the truncated alpha receptor inhibits function of the wild-type beta-receptor through formation of a ligand-dependent nonfunctional alpha beta-heterodimer. A similar heterodimer can form between the truncated beta-receptor and the wild-type alpha-receptor. These observations provide useful information for future studies using dominant negative mutations of PDGF receptors to selectively inhibit the actions of specific PDGFs in animals. PMID- 8226793 TI - FKBP-rapamycin inhibits a cyclin-dependent kinase activity and a cyclin D1-Cdk association in early G1 of an osteosarcoma cell line. AB - Upon entering a cell the natural product rapamycin, like the structurally related immunosuppressant FK506, associates with members of the FKBP family of proteins. One or more of the resulting FKBP-rapamycin complexes blocks signaling pathways emanating from some growth factor receptors. Recently, the addition of rapamycin was shown to inhibit the phosphorylation and activation of a 70-kDa ribosomal S6 protein kinase, which normally occurs minutes after the activation of certain cytokine and growth factor receptors. We now report that rapamycin can be added 4 to 6 h after the addition of serum growth factors to quiescent human osteosarcoma cells and still arrest these cells in G1. This window of action correlates with the inducible appearance of a cyclin-dependent kinase (cdk) activity, and the induction of this activity is inhibited by the addition of rapamycin. Furthermore, p36cyclin D1 associates with this cdk protein complex in lysates of untreated cells, but does not associate with this cdk protein complex in lysates of rapamycin-treated cells. Together, these studies demonstrate that FKBP rapamycin can modulate a cyclin-dependent kinase activity and a cyclin D1-cdk association during early G1 in MG-63 human osteosarcoma cells. PMID- 8226794 TI - Micelle-bound conformational preferences of a peptide derived from a murine major histocompatibility complex class I molecule. AB - Models of the micelle-bound conformation of a 17-residue major histocompatibility complex-derived peptide, [Ala85]Dk(69-85), have been determined by NMR spectroscopy and simulated annealing calculations. This peptide is a truncated, substituted version of Dk(61-85), which is a fragment of the murine major histocompatibility complex class I molecule H-2Dk. Dk(61-85) has been shown to adopt an ordered conformation required for augmentation of insulin-stimulated glucose uptake (Stagsted, J., Baase, W. A., Goldstein, A., and Olsson, L. (1991) J. Biol. Chem. 266, 12844-12847). [Ala85]Dk(69-85) retains full biological activity. Thirty-eight converged NMR structures of [Ala85]Dk(69-85) bound to dodecyl phosphocholine micelles have been generated. The NMR-derived models display a propensity for a type-I beta-bend involving residues 73-76 and an amphipathic helical region involving residues 77-84. CD spectra yield a helical content (8% at 20-25 degrees C) consistent with transient, partial helix formation. The relative orientation of the beta-bend region with respect to the helical region is not well defined by the NMR data. This may reflect true heterogeneity of the micelle-bound conformation. The NMR structures were compared with a model of [Ala85]Dk(69-85) derived from the x-ray coordinates of the human major histocompatibility complex class I allele HLA-Aw68 (Garrett, T. P. J., Saper, M. A., Bjorkmann, P. J., Strominger, T. L., and Wiley, D. C. (1989) Nature 342, 692-696). Structural features that are important for the bioactivity of [Ala85]Dk(69-85) are discussed with reference to reported structure-activity relationships (Stagsted, J., Mapelli, C., Myers, C., Matthews, B. W., Anfinsen, C. B., Goldstein, A., and Olsson, L. (1993) Proc. Natl. Acad. Sci. U.S.A., in press). A general description of the structural properties of the putative receptor site(s) that are likely to be required for binding [Ala85]Dk(69-85) is given. PMID- 8226795 TI - The SXYQRL sequence in the cytoplasmic domain of TGN38 plays a major role in trans-Golgi network localization. AB - The trans-Golgi network (TGN) plays a central role in protein sorting/targeting. TGN38 is an integral membrane protein confined mainly to the TGN. When the cytoplasmic 32-residue sequence of TGN38 was fused to the ecto- and transmembrane domains of glycophorin A (a surface protein), the resulting chimeric protein was localized to the TGN. Detailed mutagenesis of the 32-residue sequence revealed that the Ser, Tyr, and Leu residues at positions 23, 25, and 28, respectively, are essential for TGN localization. Further studies demonstrate that the sequence SXYQRL can by itself confer significant TGN localization. PMID- 8226796 TI - The structure of a bovine lung cGMP-binding, cGMP-specific phosphodiesterase deduced from a cDNA clone. AB - Polymerase chain reaction (PCR) methodology and cDNA library screening were used to isolate a cDNA clone encoding a cGMP-binding, cGMP-specific phosphodiesterase (cGB-PDE) from bovine lung. Degenerate oligonucleotides based on cGB-PDE peptide sequences were used as primers for a PCR reaction with bovine lung cDNA as the template. An 824-base pair PCR product was recovered and used as a probe to screen a bovine lung cDNA library. A 4.5-kilobase pair cDNA clone encoding a full length cGB-PDE was isolated. The open reading frame of this cDNA predicted an 875 amino acid (AA), 99,525-Da polypeptide. By Northern analysis, the cGB-PDE cDNA hybridized to a single lung 6.9-kilobase mRNA. The identity of the cGB-PDE cDNA was verified by comparison of the deduced AA sequence with several peptide sequences obtained from cGB-PDE. COS-7 cells transfected with cGB-PDE cDNA overexpressed cGMP-binding and cGMP-PDE activities characteristic of lung cGB PDE. The sequence of cGB-PDE contained a segment (AA 578-812) that was homologous to the putative catalytic region conserved among all mammalian PDEs and a segment (AA 142-526) that was homologous to the putative cGMP binding region of the cGMP stimulated PDE and the photoreceptor PDEs. As noted also for these PDEs, two internally homologous repeats were contained within the putative cGMP binding region of cGB-PDE. The amino-terminal 142 residues of cGB-PDE showed no significant homology to other PDEs and contained the serine (AA 92) which is phosphorylated by cGMP-dependent protein kinase. PMID- 8226797 TI - Identification of PECAM-1 in solid tumor cells and its potential involvement in tumor cell adhesion to endothelium. AB - PECAM-1 (CD31/EndoCAM) is an adhesion molecule in the immunoglobulin supergene family that is expressed on endothelial cells, platelets, and some hematopoietic lineage cells. In this paper, using several polyclonal and monoclonal antibodies against PECAM-1, we identified PECAM-1 molecules on human, rat, and murine solid tumor cell lines. Immunocytochemical labeling and flow cytometric analysis using either polyclonal, monoclonal, or Fab portion of the antibodies against PECAM-1 detected a distinct distribution on tumor cell surface. Immunoblotting revealed proteins ranging from 120 to 130 kDa in tumor cells derived from different species. Immunoprecipitation and subcellular fractionation studies indicated that PECAM-1 is constitutively expressed on the surface of human tumor cells (i.e. colon adenocarcinoma). The specificity of a major polyclonal anti-PECAM-1 used in the current study (i.e. SEW-3) was confirmed by the preabsorption studies. PECAM 1 molecules on tumor cells appear to bear terminal carbohydrate moieties (i.e. sialic acid residues) different from those on platelets, since neuraminidase treatment of tumor cells, unlike platelets, did not result in a mobility shift. Polymerase chain reaction (PCR) analysis of genomic DNA derived from tumor cell lines of different species revealed the presence of PECAM-1 gene in the genome. The mRNAs of PECAM-1 in tumor cells were detected by reverse transcription-PCR followed by Southern hybridization. Screening of more than 20 human, rat, and murine solid tumor cell lines indicated that PECAM-1 is widely expressed, although the level of expression varies considerably among different cell lines. The expression of PECAM-1 message in tumor cells was confirmed by Northern blotting. DNA sequencing of the PCR fragment revealed that human tumor cell PECAM 1 matches 100% to the human endothelial cell counterpart. Finally, it was demonstrated that tumor cell PECAM-1 is involved in mediating tumor cell adhesion to endothelium, as evidenced by the ability of anti-PECAM-1 antibodies to decrease the adhesion of unstimulated tumor cells to microvascular endothelial cells. PMID- 8226798 TI - Cytosolic domain of the human immunodeficiency virus envelope glycoproteins binds to calmodulin and inhibits calmodulin-regulated proteins. AB - Calmodulin (CaM), the major intracellular receptor for calcium, is involved in regulation of diverse cellular functions. Positively charged amphipathic helical segments have been identified as an important structural motif in the recognition of CaM by different CaM-activated enzymes and peptides. The carboxyl-terminal domain of the envelope glycoproteins of human and simian immunodeficiency viruses (HIV-1, HIV-2, and SIV) contain regions that can fold into amphipathic helical segments, which closely resemble the amphipathic segments found in CaM-activated enzymes. We show here that synthetic peptide analogs corresponding to the two putative amphipathic helical regions of HIV-1/WMJ gp160 bind to CaM with high affinity (Kd 31-41 nM) in the presence of calcium. They also bind CaM in the absence of calcium, although with much lower affinity. The peptides inhibit CaM regulated activation of bovine brain phosphodiesterase in vitro. The peptides also inhibit mitogen-induced lymphocyte activation, a property shared by CaM antagonists. Purified HIV-1 gp160 binds to CaM, while gp120, which lacks the putative amphipathic helical segments, does not bind CaM. In HIV-infected cells, the putative CaM-binding regions of gp160 are located intracellularly and may therefore interact with the cytosolic CaM. We postulate that CaM binding by HIV envelope proteins is likely to exert diverse modulatory effects, and the mechanism for HIV-induced cytotoxicity may involve, in part, inhibition of CaM regulated cellular functions. PMID- 8226799 TI - Molecular characterization of avian muscle titin. AB - Titin is an approximately 3000-kDa polypeptide that constitutes a set of elastic filaments that connect thick filaments to the Z-line in vertebrate striated muscle myofibrils. To characterize the primary structure of titin, three overlapping cDNA clones comprising 2.4 kilobases of avian muscle titin coding sequence were obtained from a cDNA library constructed from embryonic chick cardiac muscle RNA size-selected for large transcripts. Expression of one cDNA clone in Escherichia coli produced a fusion protein that reacted specifically with titin antibodies, and titin antiserum affinity-purified against this fusion protein reacted specifically with titin on immunoblots of chicken cardiac and skeletal muscle myofibrils. Indirect immunofluorescence localization with the fusion protein-specific antibodies demonstrated that the cDNA sequence was from the region of titin located in the myofibrillar A-band adjacent to the A/I junction. Derived amino acid sequences demonstrated a repeating pattern of fibronectin type III and immunoglobulin C2 motifs, as shown previously for a portion of rabbit skeletal muscle titin located in the central region of the A band and for other myofibrillar proteins that bind to myosin. Differences between the rabbit and chicken titin sequences included a unique, serine-rich region in one motif, which represents a potential phosphorylation site. This is the first report of sequence information for avian titin from a previously uncharacterized portion of the titin molecule. PMID- 8226800 TI - A somatic cell mutant defective in phosphatidylglycerophosphate synthase, with impaired phosphatidylglycerol and cardiolipin biosynthesis. AB - Phosphatidylglycerophosphate (PGP) synthase catalyzes a reaction involved in the synthesis of phosphatidylglycerol (PG), which serves as a metabolic precursor for cardiolipin (CL), found primarily in the mitochondrial membranes of eukaryotic cells. We isolated a Chinese hamster ovary cell mutant (designated PGS-S) with a specific lesion in PGP synthase by using an in situ enzymatic assay for the enzyme. This mutant was obtained by introducing a second mutation into mutant PGS P that had been generated by first-step mutagenesis. The PGP synthase activities in cell extracts of mutant PGS-S grown at 33 and 40 degrees C were 14 and 1% of those in the wild type cells, respectively; in addition, PGP synthase in cell extracts of mutant PGS-S exhibited higher sensitivity to heat than that of the wild type. Mutant PGS-S also showed a temperature-dependent defect in the synthesis of PG and CL in vivo, together with temperature sensitivity for cell growth. A temperature-resistant revertant of mutant PGS-S simultaneously restored PGP synthase activity and the ability to synthesize PG and CL in vivo to nearly the same levels as those of mutant PGS-P. These results constitute genetic evidence that PGP synthase is responsible for PG synthesis and is essential for cell growth. PMID- 8226801 TI - Mitochondrial dysfunction of a cultured Chinese hamster ovary cell mutant deficient in cardiolipin. AB - In our preceding paper, we reported that a temperature-sensitive Chinese hamster ovary cell mutant, PGS-S, with thermolabile phosphatidylglycerophosphate synthase was defective in the biogenesis of both phosphatidylglycerol and cardiolipin (CL) at a nonpermissive temperature (Ohtsuka, T., Nishijima, M., and Akamatsu, Y. (1993) J. Biol. Chem. 268, 22908-22913). To investigate the biological role of cardiolipin, we examined the structure and function of mitochondria in mutant PGS S cells, since CL is primarily found in the mitochondrial membranes of eukaryotic cells. Under conditions where the formation of CL was impaired, this mutant had both morphological and functional mitochondrial abnormalities, manifested by more stringent temperature sensitivity for cell growth in glucose-deficient medium and by reduced ATP production, increased glycolysis, and reduced oxygen consumption in intact cells. Rotenone-sensitive NADH oxidase activity in cell extracts was also reduced in the mutant cultivated at a nonpermissive temperature, showing a defect(s) in the respiratory electron transport chain of mitochondria. Of the respiratory chain complexes, rotenone-sensitive NADH-ubiquinone reductase (Complex I) was most severely impaired in the mutant, whereas its activity was restored in a revertant of the mutant that had regained the ability to synthesize CL. These results suggest that CL plays a critical role in mitochondrial functions, at least in the respiratory electron transport chain. PMID- 8226802 TI - DNA topoisomerase II and casein kinase II associate in a molecular complex that is catalytically active. AB - Immunoprecipitation of DNA topoisomerase II from yeast results in a preparation that contains casein kinase II; this suggests that the two proteins may associate in the intact cell. Purified recombinant topoisomerase II and casein kinase II associate to form a complex in vitro which is stable after topoisomerase II becomes phosphorylated by the kinase. Studies with isolated recombinant casein kinase II subunits disclosed that although the alpha (catalytic) subunit alone can efficiently phosphorylate topoisomerase II, the formation of a stable topoisomerase II-casein kinase II association requires the presence of the beta subunit of the kinase. Both proteins engaged in this complex retain their catalytic activities. Naturally occurring polyamines and polyanionic compounds appear to be crucial factors governing the interaction between the two proteins. Although the biological significance of a stable catalytically active topoisomerase II-casein kinase II molecular complex remains to be defined, these observations suggest the possibility of a novel mechanism regulating topoisomerase II and casein kinase II activities. PMID- 8226803 TI - Activation of the zymogen of hepatocyte growth factor activator by thrombin. AB - Hepatocyte growth factor activator (HGF activator) is a serine protease which converts single-chain HGF to the active two-chain form. HGF activator purified from human serum has a molecular mass of 34 kDa and consists of two chains held together by a disulfide bond. The nucleotide sequence of HGF activator cDNA shows that HGF activator is derived from the COOH-terminal region of a precursor of 655 amino acids by proteolytic cleavage of the bonds between Arg372 and Val373 and between Arg407 and Ile408 and that the precursor consists of multiple domains homologous to those observed in blood coagulation factor XII. In this study, we identified the precursor of HGF activator in human plasma using an enzyme-linked immunosorbent assay system. The precursor was purified from plasma by a five-step procedure. The purified precursor did not activate single-chain HGF. The precursor was efficiently cleaved in vitro by thrombin, at the bond between Arg407 and Ile408, in the presence of negatively charged substances. The cleaved precursor activated single-chain HGF. These findings led us to conclude that HGF activator is present in plasma as an inactive zymogen and that the zymogen is activated by the cleavage of the bond between Arg407 and Ile408 by thrombin. Characteristic structural domains in the NH2-terminal region of the zymogen may be involved in the binding of the zymogen to negatively charged substances, which stimulates the activation of the zymogen by thrombin. PMID- 8226804 TI - Glucocorticoid inhibits thrombin-induced expression of platelet-derived growth factor A-chain and heparin-binding epidermal growth factor-like growth factor in human aortic smooth muscle cells. AB - Proliferation of smooth muscle cells (SMCs) in atherosclerosis may be modulated by several growth regulatory molecules. At least two mitogens for SMCs, platelet derived growth factor (PDGF) A-chain and heparin-binding epidermal growth factor like growth factor (HB-EGF), can be produced by SMCs themselves and may stimulate smooth muscle proliferation in an autocrine or paracrine fashion. We examined the effects of thrombin, which may be generated at the site of vascular injury during atherogenesis, and the potent anti-inflammatory glucocorticoid, dexamethasone (DEX), on the expression of the genes encoding these two growth factors. Since both PDGF A-chain and HB-EGF have affinity for heparin, we also examined the effect of thrombin and DEX on the release of heparin binding mitogenic activity from SMCs. Treatment of SMCs with thrombin resulted in increases both in the level of the PDGF-A and HB-EGF transcripts in the cells, as well as in released heparin-binding growth factor activity. DEX inhibits the thrombin-stimulated release of mitogenic activity in a dose-dependent manner. An enzyme-linked immunoadsorbent assay showed that DEX inhibits both constitutive and thrombin stimulated release of PDGF-AA. DEX also decreases both constitutive and thrombin stimulated mRNA levels for PDGF A-chain and HB-EGF and destabilizes the transcripts for both growth factors. A nuclear run-on assay revealed that DEX acts, in addition, to inhibit constitutive and thrombin-stimulated transcription of the PDGF A-chain and HB-EGF genes. Thus, these findings indicate that expression of PDGF A-chain and HB-EGF may be regulated by thrombin and glucocorticoid at the transcription level. Our results are consistent with the involvement of thrombin-induced growth factor expression in neointimal SMC proliferation and suggest the possibility that intimal proliferation may be attenuated by glucocorticoids. PMID- 8226805 TI - Purification of a novel ras GTPase-activating protein from rat brain. AB - GTPase-activating protein (GAP) and neurofibromin, a gene product of neurofibromatosis type I gene, have been identified as factors that stimulate GTPase activity of ras p21. We have previously suggested the presence of novel GAP activity that is immunologically distinguishable from GAP or neurofibromin in both the cytosolic and the particulate fractions of rat brain (Hattori, S., Maekawa, M., and Nakamura, S. (1992) Oncogene 7, 481-485). We have purified this novel GAP molecule from the cytosolic fraction of rat brain by more than 200,000 fold by five successive column chromatographies with a recovery of 6%. Apparent molecular mass of this molecule was estimated to be 100 kDa (p100GAPras). The same p100GAPras was purified from the particulate fraction after extraction with high salt. The activation of GTPase was observed with normal ras p21 but not with oncogenic ras p21, Rap1B/smg21B, or Ram25K. The dissociation constant of p100GAPras toward ras p21 estimated by competitive inhibition using ras p21 in complex with nonhydrolyzable analog of GTP was two times higher than that of neurofibromin and was lower than that of GAP by 2 orders of magnitude. These results clearly indicate that p100GAPras is a novel ras GAP molecule. PMID- 8226806 TI - Poly(A) tail metabolism and function in eucaryotes. PMID- 8226807 TI - Activation of protein kinase C inhibits cellular production of the amyloid beta protein. AB - The 39-43-amino acid amyloid beta-protein (A beta), which is progressively deposited in cerebral plaques and blood vessels in Alzheimer's disease (AD), is released by cultured human cells during normal metabolism. Here we show that agents which activate protein kinase C or otherwise enhance protein phosphorylation caused a substantial decrease in A beta production in vitro. Protein kinase C activation also markedly decreased A beta release from cells that express mutant forms of the beta-amyloid precursor protein genetically linked to familial AD. Inhibition of A beta secretion could also be effected by direct stimulation of m1 muscarinic acetylcholine receptors with carbachol. These results demonstrate that activation of the protein kinase C signal transduction pathways down-regulates the generation of the amyloidogenic A beta peptide. Pharmacologic agents that activate this system, including a variety of first messengers, could potentially slow the development or growth of some A beta plaques during the early stages of AD. PMID- 8226808 TI - Identification of Trk binding sites for SHC and phosphatidylinositol 3'-kinase and formation of a multimeric signaling complex. AB - Phosphotyrosine-containing synthetic peptides were used to identify the binding sites for cellular polypeptides involved in nerve growth factor receptor/Trk mediated signal transduction. In vitro association of SHC and the p85 subunit of phosphatidylinositol 3'-kinase with the Trk tyrosine kinase was prevented only by phosphorylated Y-490- and Y-751-containing peptides, respectively. In spite of the close proximity of the p85 binding site to that of phospholipase C gamma (Y 785), both target proteins are able to interact with the same receptor molecule simultaneously. PMID- 8226809 TI - Proteolysis and lipid-facilitated translocation are distinct but competitive processes that regulate secretion of apolipoprotein B in Hep G2 cells. AB - Under lipid-poor conditions, most newly synthesized apolipoprotein B100 (apoB) undergoes rapid degradation in Hep G2 cells such that only a small fraction of newly synthesized apoB is actually secreted. Addition of oleate to Hep G2 culture medium stimulates apoB secretion by a post-translational mechanism. In the current studies we have explored oleate-stimulation of apoB secretion by using calpain inhibitor I, N-acetyl-leucyl-leucyl-norleucinal (ALLN), a compound that inhibits the intracellular degradation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase and the T cell receptor alpha subunit. Preincubation of Hep G2 cells with ALLN (40 micrograms/ml) for 1 h markedly inhibited degradation of newly synthesized apoB. Whereas only 32% of newly labeled apoB remained intact (cells+medium) in control cells after a 10-min pulse with [3H]leucine followed by a 60-min chase, 84% of labeled apoB was intact in ALLN-treated cells. However, most of the ALLN-protected apoB remained intracellular, as ALLN did not stimulate the rate of apoB secretion over the control rate (12 versus 9.2%). Although secretion of apoB was not accelerated, the protection afforded by ALLN continued for several hours, and labeled apoB continued to be secreted over 3 h of chase after which secretion ceased. The protection afforded by ALLN resulted in 37% of labeled apoB secreted by 3 h compared to 15% in control cells. In contrast, simultaneous treatment of cells with ALLN and oleate both accelerated and increased total apoB secretion, such that 36% of initially labeled apoB was recovered in the medium by 60 min and 71% of labeled apoB was secreted by 180 min of chase. These data show that ALLN and oleate affect apoB metabolism by different mechanisms. Although ALLN can protect nascent apoB from rapid early intracellular degradation, it does not accelerate apoB secretion. In contrast, although our results can not rule out the possibility that oleate may directly inhibit proteolysis of apoB, oleate appears to protect apoB mainly by facilitating transport of apoB out of a protease-containing compartment associated with the endoplasmic reticulum. PMID- 8226810 TI - Receptor activation of and signal generation by the lutropin/choriogonadotropin receptor. Cooperation of Asp397 of the receptor and alpha Lys91 of the hormone. AB - We have developed a novel method of reciprocal substitution mutation to identify pairing of amino acids within a receptor and its ligand. Using this method, we demonstrate for the first time that a pair of counterionic amino acids, one from the lutropin/choriogonadotropin receptor and the other from the ligand (human choriogonadotropin), cooperate and perhaps interact with each other to activate the receptor and to generate hormonal signal. In this study, Asp397 of the receptor was converted to Lys while Lys91 of the alpha subunit of human choriogonadotropin was substituted with Asp, thus maintaining the counterionic nature of this pair of amino acids. Mutation at each of these positions does not affect the hormone-receptor interaction but results in the significant or complete loss of the bioactivity of both the receptor and the hormone. However, when the impotent mutant receptor and mutant hormone were paired to interact together, they induced cAMP synthesis, resulting in a potent receptor-hormone couple. Substitutions with other amino acids that eliminated the counterionic nature failed to induce cAMP synthesis. Our results shed light on the molecular mechanisms of receptor activation and will serve as a model for other G-protein coupled peptide receptors, particularly glycoprotein hormone receptors. PMID- 8226811 TI - Peptide environment specifies conformation. Helicity of hydrophobic segments compared in aqueous, organic, and membrane environments. AB - Transmembrane segments in integral membrane proteins exist characteristically as helices in lipid bilayers, yet are often rich in residues considered helix destabilizing (Val, Ile, Gly) in soluble proteins. We propose that helicity of a transmembrane segment is likely to be affected by factors other than the "intrinsic" helical propensities of its component amino acids. This hypothesis is tested by comparing the conformation(s) in aqueous, organic, membrane-mimetic (micellar), and membrane (bilayer) environments of designed model peptides with systematically altered helical propensity and/or segmental hydrophobicity. Peptides of prototypic sequence NH2-(Ser-Lys)2-Ala5-Leu6-Ala7-Ala8-Leu9-Ala10-++ +Trp11-Ala12-Leu13-Ala14- (Lys-Ser)3-OH were synthesized, which incorporate a hydrophobic core "guest" segment (residues 5-14) into a water-soluble hydrophilic host matrix. Related peptides featured substitution of Leu6,9,13-->Gly, Leu6,9,13 ->Ala, and Ala7,10,14-->Gly. Circular dichroism spectra revealed that algorithms for soluble proteins correctly predicted peptide helical proclivities in aqueous solutions, but peptide helicity in organic (trifluoroethanol) solvents, membrane mimetic SDS micelles, and negatively charged lipid bilayer vesicles, was found to be governed almost exclusively by the segmental hydrophobicity of the peptide mid hydrophobic core segment. In related Trp fluorescence studies, peptide-membrane association was similarly correlated with extent of hydrophobic interaction. PMID- 8226812 TI - The acceptor substrate specificity of porcine submaxillary UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase is dependent on the amino acid sequences adjacent to serine and threonine residues. AB - The acceptor substrate specificity of a pure polypeptide N acetylgalactosaminyltransferase has been examined with synthetic polypeptides with sequences identical, or similar to those found in porcine mucin or human erythropoietin. The sequences adjacent to either threonine or serine markedly influence the formation of GalNAc-O-Thr and GalNAc-O-Ser. Examination of the mucin-like peptide VLGXXAV, where X is Thr, Ser, or Ala, shows only Thr containing peptides to be acceptors. The best substrate is formed when XX is TT. Peptides with XX as either AT or TA are less effective and those with XX as either ST or TS are much less effective acceptors. The amino acids adjacent to serine in the peptide formed by residues 121-131 in human erythropoietin, PPDAASAAPLR, also markedly influence the formation of GalNAc-O-Ser. Thus, PPDASSSAPLR and PPDVVSVVPLR are about 5- and 30-fold, respectively, less active than the erythropoietin peptide. The peptide PPDGGSGGPLR is inactive. The shorter peptide DAASAAPL is also about 5-fold less active than the full-length peptide, but the peptide AASAA is inactive. These studies indicate that one transferase can form both GalNAc-O-Ser and GalNAc-O-Thr residues when the sequences adjacent to the glycosylated residue are of the proper kind. Thus, in contrast to earlier suggestions, there is no evidence that different transferases form GalNAc-O-Ser and GalNAc-O-Thr. Examination of tissue homogenates from various tissues confirms this conclusion. PMID- 8226813 TI - Comparison of the membrane binding kinetics of bovine prothrombin and its fragment 1. AB - Total internal reflection fluorescence microscopy has been used to compare the membrane binding characteristics of fluorescein-labeled bovine prothrombin and fluorescein-labeled bovine prothrombin fragment 1. The Ca(2+)-dependent association of these proteins with quartz-supported planar membranes composed of mixtures of phosphatidylserine (2-10 mol%) and phosphatidylcholine was examined. Equilibrium binding measurements showed that the apparent equilibrium dissociation constants increased with decreasing molar fractions of phosphatidylserine and that the dissociation constants were somewhat lower for intact prothrombin. Kinetic measurements, using fluorescence photobleaching recovery, showed that the measured dissociation rates were approximately equivalent for prothrombin and fragment 1 and did not change with the protein solution concentration or the molar fraction of phosphatidylserine. The kinetic data also implied that the surface binding mechanism for both proteins is more complex than a simple reversible reaction between monovalent proteins and monovalent surface sites. Measured equilibrium and kinetic constants are reported and compared for prothrombin and fragment 1 on planar membranes. PMID- 8226814 TI - Effect of heavy chain signal peptide mutations and NH2-terminal chain length on binding of anti-digoxin antibodies. AB - In certain instances, antibody variable region mutations outside of the antigen combining site influence antigen binding. We reported previously that a heavy chain mutation (Ser-94-->Arg) decreased binding of the anti-digoxin antibody 40 150, whereas an additional signal peptide mutation at the -2 position (Gln-->Pro) causing NH2-terminal 2-residue truncation partially restored binding. To assess the combined effects on binding of two seemingly distant mutations, we constructed signal peptide mutations and NH2-terminal deletions in the presence of Ser-94 and Arg-94. Deletions of one to three amino acids had little effect on binding for Ser-94 mutants, whereas 2-residue truncations produced directly or by signal peptide mutation increased affinity approximately 40-fold for Arg-94 mutants. These observations are consistent with the reported computer-generated model of antibody 40-150. Introduction of Pro at the signal peptide -3 position in 40-150 resulted in cleavage at alternative sites, with varying effects on affinity. Introduction of Pro at -2 into the anti-digoxin antibody 26-10 resulted, unexpectedly, in expression of heavy chains with 3 extra NH2-terminal residues, causing an approximately 100-fold reduction in affinity. Thus, both extensions and deletions of the heavy chain amino terminus can enhance or reduce antigen binding, depending on the structural context of specific antigen combining sites. PMID- 8226815 TI - Conformation of apolipoprotein E in lipoproteins. AB - Lysine (Lys) residues in apolipoprotein (apo) E are known to be involved in binding of apoE-containing lipoproteins to the low density lipoprotein (LDL) receptor. To examine the microenvironments of the Lys residues of apoE-3 in a variety of lipid-associated states, we have used a high resolution 13C-NMR method in which Lys were reductively methylated with [13C] formaldehyde. Over a wide pH range, the spectrum of apoE in canine HDLc, a spherical lipoprotein particle, exhibited two peaks from Lys epsilon-amino groups. The two pools of Lys in HDLc titrate with pK alpha values of 10.4 ("normal") and 9.3 ("active"). In contrast, eight epsilon-[13CH3]2Lys peaks (delta = 42.5-44.5 parts/million) with pK alpha values ranging from 8.2 to 10.1 and 7.8 to 10.5 were observed at pH 9.5 for human and canine apoE, respectively, in discoidal complexes with dimyristoyl glycerophosphocholine. A single Lys microenvironment was observed for apoE present in a disordered, lipid-free, state in 8 M urea, confirming the fact that the lipid environment is modulating the conformation of apoE. The above data demonstrate that the conformation of apoE, as reflected by the Lys microenvironments, on spherical HDLc particles is different from that on discoidal complexes. PMID- 8226816 TI - 4S-limonene synthase from the oil glands of spearmint (Mentha spicata). cDNA isolation, characterization, and bacterial expression of the catalytically active monoterpene cyclase. AB - The committed step in the biosynthesis of monoterpenes in mint (Mentha) species is the cyclization of geranyl pyrophosphate to the olefin (-)-4S-limonene catalyzed by limonene synthase (cyclase). Internal amino acid sequences of the purified enzyme from spearmint oil glands were utilized to design three distinct oligonucleotide probes. These probes were subsequently employed to screen a spearmint leaf cDNA library, and four clones were isolated. Three of these cDNA isolates were full-length and were functionally expressed in Escherichia coli, yielding a peptide that is immunologically recognized by polyclonal antibodies raised against the purified limonene synthase from spearmint and that is catalytically active in generating from geranyl pyrophosphate a product distribution identical to that of the native enzyme (principally limonene with small amounts of the coproducts alpha- and beta-pinene and myrcene). The longest open reading frame is 1800 nucleotides and the deduced amino acid sequence contains a putative plastidial transit peptide of approximately 90 amino acids and a mature protein of about 510 residues corresponding to the native enzyme. Several nucleotide differences in the 5'-untranslated region of all three full length clones suggest the presence of several limonene synthase genes and/or alleles in the allotetraploid spearmint genome. Sequence comparisons with a sesquiterpene cyclase, epi-aristolochene synthase from tobacco, and a diterpene cyclase, casbene synthase from castor bean, demonstrated a significant degree of similarity between these three terpenoid cyclase types, the first three examples of this large family of catalysts to be described from higher plants. PMID- 8226817 TI - Selection of targeted biological modifiers from a bacteriophage library of random peptides. The identification of novel calmodulin regulatory peptides. AB - The interaction of short amino acid sequences is the basis of molecular recognition and biological regulation in many cellular systems. Libraries of random peptides provide an approach to identify peptides that can be used to modulate, in a targeted fashion, the function of specific gene products. We have used a library of random peptides designed and constructed in the M13 bacteriophage to select calcium-dependent calmodulin binding-peptides. Twenty eight independent sequences were obtained; all contained a tryptophan within the fifteen-amino acid insert. In 11 sequences, the tryptophan was located in the first possible variable position of the inserted sequence and was followed by a proline. The tryptophan-proline combination was also present in six additional isolates but at various other positions within the peptide insert. Synthetic peptides, representative of the calmodulin binding sequences, bound to calmodulin in a calcium-dependent fashion, competed with known calmodulin inhibitors and, when introduced via a patch pipette, inhibited calcium-activated chloride conductance of the colonic epithelial cell line, T84. This report demonstrates the utility of identifying modifiers of biological function and should prove to be a valuable approach in understanding the cellular role of proteins of unknown function. PMID- 8226818 TI - Oxygen equilibrium studies of cross-linked iron-cobalt hybrid hemoglobins. Models for partially ligated intermediates of cobalt hemoglobin. AB - To probe the molecular mechanism of allosteric function of cobaltous protoporphyrin-substituted hemoglobin (CoHb), a series of alpha, alpha-cross linked symmetric and asymmetric Fe-Co hybrid hemoglobins, which contain (1Co porphyrin/3Fe porphyrins), (2Co porphyrins/2Fe porphyrins), and (3Co porphyrins/1Fe porphyrin) per tetramer, have been prepared. Because only Fe porphyrin-containing subunits react with CO, these Fe-Co hybrids are converted to mono-, di-, and tri-CO-ligated states in the presence of CO, respectively, and are proposed to stand as models for mono-, di-, and tri-ligated intermediates of CoHb, respectively. The oxygen binding properties of these Fe-Co hybrids were investigated by measuring oxygen binding isotherms in the presence of CO as a function of pH in the presence and absence of IHP. The ligation of CO to a beta subunit causes larger changes in the oxygen affinity and the Bohr effect than that to an alpha subunit, indicating that the ligation to a beta subunit induces larger affinity-related structural changes in cross-linked CoHb. Di- and tri-CO ligated intermediates exhibited substantially increased oxygen affinity, reduced Bohr effect, and reduced IHP effect, indicating that they are in high affinity states. Calculation of the Adair equilibrium constants for the first and last oxygenation steps for each of these intermediates permitted the determination of the level of free energy of cooperation. The intermediately ligated species of cross-linked CoHb are distributed in multiple levels of free energy of cooperation within the free energy difference of 1.14 kcal mol-1 between deoxy and fully ligated states of cross-linked CoHb at pH 7.4. The ligation process in CoHb is determined by the number and distribution of the bound ligands, and ligation takes place through steps that require minimal free energy changes. PMID- 8226819 TI - Protein kinase C-mediated bidirectional regulation of DNA synthesis, RB protein phosphorylation, and cyclin-dependent kinases in human vascular endothelial cells. AB - In human umbilical vein endothelial cells, activators of protein kinase C (PKC) exert cell cycle-dependent, bidirectional growth regulatory effects. Thus, phorbol 12,13-dibutyrate or 1,2-dioctanoylglycerol potentiates growth factor induced DNA synthesis up to 3-fold when they act during the early G1 phase, whereas they completely inhibit the initiation of DNA synthesis when they act in the late G1 phase. In addition, the PKC activators induce a rapid inhibition of the ongoing DNA synthesis when they are applied after entry into the S phase. The effects of the PKC activators in both stimulatory and inhibitory directions are abolished in PKC-downregulated cells. The cell cycle-dependent, PKC-mediated bidirectional growth regulation is closely associated with either potentiation or inhibition of RB protein phosphorylation and the histone H1 kinase activity of cyclin-dependent kinases (cdks) cdc2 and cdk2, which normally accumulate along the G1 to the S phase transition. Northern and Western blot analyses of cdc2 and cdk2 have revealed that PKC regulates the cdks at multiple steps in distinct ways. Thus, for cdc2, the levels of mRNA and protein as well as the extent of post-translational modification are all subject to the PKC-mediated regulation. In contrast, the level of mRNA or protein of cdk2 is not affected by PKC stimulation at any phase of the cell cycle. These results demonstrate the existence of a complex array of PKC-cdk signaling pathways, which mediate temporally organized bimodal growth regulation in endothelial cells. PMID- 8226820 TI - Evaluation of activity of putative superoxide dismutase mimics. Direct analysis by stopped-flow kinetics. AB - By stopped-flow kinetic analysis, we have directly evaluated the superoxide dismutase (SOD) activity of a number of organic nitroxides and iron- and manganese-based complexes that have been attributed with having SOD activity based upon competition experiments with cytochrome c. In 60 mM HEPES buffer, pH 8.1, or 50 mM potassium phosphate buffer, pH 7.8, Mn(II) and manganese complexes of desferal had no detectable SOD activity by stopped-flow analysis (catalytic rate constant (kcat) < 10(5.5) M-1 s-1), whereas Mn(II) and manganese complexes of desferal inhibited the reduction of cytochrome c by superoxide generated by the xanthine/xanthine oxidase system. Fe(II)-N,N,N',N'-tetrakis(2 pyridylmethyl)ethylenediamine (FeTPEN) was eight times more active than Fe(III) tris[N-(2-pyridylmethyl)-2-aminoethyl]amine(Fe-TPAA) in the cytochrome c assay, but only FeTPAA catalyzed the first-order decay of superoxide (kcat = 2.15 x 10(6) M-1 s-1) by stopped-flow. Fe(III)-tetrakis(4-N-methylpyridyl)porphine (FeTMPP) was active at low micromolar concentrations in both the cytochrome c and stopped-flow assays. At high micromolar concentrations, the organic nitroxides 2,2,6,6-tetramethylpiperidin-1-yloxy (TEMPO) and 4-hydroxy-2,2,6,6 tetramethylpiperidin-1-yloxy (TEMPOL) were inhibitory in the cytochrome c assay, but showed no detectable SOD activity by stopped-flow. None of the tested compounds inhibited xanthine oxidase activity as shown by the measurement of urate production. Under the conditions of the cytochrome c assay, FeTPEN, TEMPO, and TEMPOL oxidized reduced cytochrome c which rationalizes the false positives for these compounds in this assay. The inhibitory activities of Mn(II) and the manganese desferal complexes in the cytochrome c assay appear to be due to a stoichiometric, not catalytic, reaction with superoxide as catalytic amounts of these agents do not induce a first-order decay of superoxide as shown by stopped flow. PMID- 8226821 TI - A single residue, aspartic acid 95, in the delta opioid receptor specifies selective high affinity agonist binding. AB - The enkephalins, dynorphins, and endorphins are endogenous opioids which function as neurotransmitters, neuromodulators, and hormones and are involved in the perception of pain, modulation of behavior, and regulation of autonomic and neuroendocrine function. Pharmacological studies have defined three classes of opioid receptors, designated as delta, kappa, and mu. To investigate mechanisms by which agonists and antagonists interact with the delta opioid receptor, we have substituted aspartic acid 95 in the transmembrane segment 2 of the cloned mouse delta opioid receptor with an asparagine (D95N). The D95N mutant receptor had reduced affinity for delta receptor-selective agonists such as enkephalin, [D Pen2,D-Pen5]enkephalin and [D-Ser2,Leu5]enkephalin-Thr6 such that it did not bind these peptides even at micromolar concentrations. The binding of delta-selective non-peptide agonists was also reduced. In contrast, the delta receptor-selective antagonists, such as naltrindole, the benzofuran analog of naltrindole, and 7 benyllidenenaltrexone, bound equally well to the wild-type and mutant receptor. Similarly, non-selective opioid agonists such as bremazocine and buprenorphine, which interact with delta, kappa, and mu opioid receptors, showed no difference in binding to the wild-type and mutant delta receptor. The D95N mutant remained coupled to G proteins, and the receptor was functionally active since it mediated agonist inhibition of cAMP accumulation. These results indicate that selective agonists and antagonists bind differently to the delta receptor and show that Asp 95 contributes to high affinity delta-selective agonist binding. The identification of a key residue involved in selective agonist binding to the delta opioid receptor will facilitate the development of novel therapeutic reagents that can be used for the treatment of chronic pain and other conditions. PMID- 8226822 TI - Phosphorylation of the inositol trisphosphate receptor in isolated rat hepatocytes. AB - The inositol trisphosphate receptor (IP3R) in brain has been shown to be a substrate for several different protein kinases in vitro. We have studied the phosphorylation of the IP3R in intact cells by using isolated hepatocytes and an antibody to immunoprecipitate the receptor protein from detergent extracts. Stimulation of 32P-labeled hepatocytes with glucagon or N6,2'-O dibutyryladenosine 3',5'-cyclic monophosphate (db-cAMP) markedly increased phosphorylation of the IP3R. However, no increase was observed in response to angiotensin II, vasopressin, 12-O-tetradecanoyl-phorbol-13-acetate, or epidermal growth factor. The kinetics of phosphorylation in response to glucagon was both rapid and transient. In agreement with previous studies, physiological concentrations of Ca2+ stimulated D-myo-inositol 1,4,5-trisphosphate (IP3) binding to permeabilized hepatocytes (Pietri, F., Hilly, M., and Mauger, J.-P. (1990) J. Biol. Chem. 265, 17478-17485). Pretreatment of cells with db-cAMP had no effect on binding in the absence of added Ca2+ but enhanced binding measured in the presence of basal low concentrations (0.16-0.25 microM) of Ca2+ and decreased the concentration of Ca2+ required for half-maximal stimulation. The effect of db-cAMP was associated with an increase in affinity of the IP3 binding site without a change in maximum number of binding sites. Preincubation of intact hepatocytes with okadaic acid alone produced an increase in basal phosphorylation of the IP3R, and maximal phosphorylation of the receptor was observed in the presence of both okadaic acid and db-cAMP. However, okadaic acid blocked the effect of db-cAMP and inhibited the effect of Ca2+ on IP3 binding. Detergent solubilized binding sites were already fully activated and insensitive to modulation by Ca2+ or cAMP-dependent protein kinase. It is proposed that the receptor in native membranes is inhibited and that Ca2+ and cAMP-dependent protein kinase may act to relieve this inhibition. PMID- 8226823 TI - Inhibition of vascular endothelial cell growth by activin-A. AB - The demonstration of type 2 activin receptor expression in human umbilical vein endothelial cells prompted an investigation of the effects of the activin/inhibin family of hormones on vascular endothelial cell growth. Recombinant activin-A inhibited [3H]methylthymidine uptake and growth of a panel of endothelial cell types; recombinant inhibin-A was without effect. Affinity cross-linking studies demonstrated the presence of type 1 and type 2 activin receptors on the surface of bovine aortic endothelial cells, while detailed analysis of type 2 activin receptor expression revealed both type 2 and type 2B activin receptor mRNA in all endothelial cell types analyzed. In addition, capillary endothelial cells were found to express activin-beta A subunit mRNA and protein, the levels of which were increased in response to transforming growth factor (TGF)-beta. Furthermore, activin-A and TGF-beta caused additive inhibition of capillary endothelial cell [3H]methylthymidine uptake. These findings implicate the activins in the regulation of endothelial cell function, and suggest that TGF-beta and activin may interact to inhibit capillary endothelial cell growth. PMID- 8226824 TI - Differences in the regulation of protein synthesis, cyclin B accumulation, and cellular growth in response to the inhibition of DNA synthesis in Chinese hamster ovary and HeLa S3 cells. AB - We have shown previously that there are significant differences between mammalian cell lines in response to disruption of the assembly of the mitotic spindle apparatus (Kung, A. L., Sherwood, S. W., and Schimke, R. T. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 9553-9557). In this paper we report that there are also significant differences between mammalian cell lines in response to the inhibition of DNA synthesis. In HeLa S3 cells protein synthesis is down regulated, and cellular growth is arrested in response to the inhibition of DNA synthesis. Upon release from inhibition and resumption of normal growth, cellular viability is maintained near untreated control levels. In contrast, Chinese hamster ovary cells continue to accumulate protein and continue to undergo cellular growth during the period of DNA synthesis inhibition. Cyclin B levels accumulate throughout the period of inhibition and rapidly exceed normal levels at mitosis. The degree of aberrant growth during the period of transient DNA synthesis inhibition is directly related to the degree of subsequent cytotoxicity. If protein accumulation and cellular growth are limited with partially inhibitory levels of cycloheximide during the period of DNA synthesis inhibition, the cytotoxic effects are abolished. These results support the concept that aberrant growth and accumulation of proteins during a transient period of DNA synthesis inhibition are primary determinants of subsequent cell killing. PMID- 8226825 TI - Biosynthesis of the side chain of ubiquinone:trans-prenyltransferase in rat liver microsomes. AB - The trans-prenyltransferase activity present in rat liver microsomes was investigated using an in vitro system. Geranyl-PP, but not farnesyl-PP is utilized as substrate. The pH optimum is at 8.0, and Mn2+ and Mg2+ activate, while Zn2+ completely inhibits the enzyme. Digitonin, taurodeoxycholate, and Tween 80 increase this activity, whereas deoxycholate and SDS are inhibitory. In contrast to the cis-prenyltransferase, the trans-prenyltransferase is not dependent on cytosolic protein factors. The trans-prenyltransferase is present at the cytoplasmic surface of rough and smooth microsomes. Solanesyl-PP and all trans-geranylgeranyl-PP inhibit the transferase activity but poly-cis-polyprenyl 12-PP, an intermediate in the cis-prenyltransferase reaction, does not. The enzyme reaction gives rise to two products, solanesyl-PP and an unidentified polyprenyl metabolite. Mevinolin treatment distinguishes this enzyme from the cytoplasmic geranylgeranyl-PP synthase. The results demonstrate that rat liver microsomes synthesize solanesyl-PP via a trans-prenyltransferase, which is distinct from cis-prenyltransferase and geranylgeranyl-PP synthase. PMID- 8226826 TI - Retinoic acid biphasically regulates the gene expression of hepatitis B virus surface antigen in human hepatoma Hep3B cells. AB - We used human hepatoma Hep3B/C16 cells as a model to examine the effect of all trans retinoic acid on the gene expression of hepatitis B surface antigen (HBsAg). Hep3B/C16 is a clonal derivative of human hepatoma Hep3B cell which was stably transfected with HBsAg DNA sequences and can produce hepatitis B virus surface antigen. We analyzed the HBsAg product and mRNA in Hep3B/C16 cells which were exposed to retinoic acid for different periods of time. The level of HBsAg started to increase after 24 h and reached maximum at 48 h of retinoic acid treatment. However, the level of HBsAg expression was severely suppressed compared to the control cells after long term (120 h) retinoic acid treatment. Such biphasic regulation of HBsAg production by retinoic acid was paralleled by the changes of HBsAg mRNA. Nuclear run-on assays also demonstrated that the retinoic acid-mediated regulation was determined at least in part at the transcriptional level. Furthermore, an exposure of the cells to retinoic acid for only 8 h was sufficient to show that up- and down-regulation of HBsAg gene occurred 2 and 5 days later. Using a transient expression system, we demonstrated that the retinoic acid response element is located within the 5'-flanking region of the HBsAg gene. PMID- 8226827 TI - High density lipoprotein conversion mediated by human plasma phospholipid transfer protein. AB - Phospholipid transfer protein (PLTP) was purified from lipoprotein-free human plasma, obtained upon treatment of plasma with dextran sulfate and Ca2+, by employing a series of column chromatography. Upon sodium dodecyl sulfate polyacrylamide gel electrophoresis, the purified PLTP showed a single main band, corresponding to the molecular mass of 78 kDa. However, isoelectric focusing of the purified preparation gave multiple bands with pI ranging from 4.3 to 5.1, indicative of microheterogeneity. Purified PLTP was shown to possess not only phospholipid transfer activity, but also high density lipoprotein (HDL) conversion activity (Tu, A.-Y., Nishida, H. I., and Nishida, T. (1990), FASEB J. 4, A2148; Jauhiainen, M., Metso, J., Pahlman, R., Blomqvist, S., van Tol, A., and Ehnholm, C. (1993) J. Biol. Chem. 268, 4032-4036). Isolated HDL3 was enlarged to the size of HDL2b upon incubation with purified PLTP for 6 h at 37 degrees C at the PLTP/HDL3 molar ratio of approximately 1:45. Both the HDL conversion and the phosphatidylcholine transfer activities of purified PLTP were effectively inhibited by rabbit anti-PLTP immunoglobulin G. The primary importance of PLTP in the HDL enlargement that occurs in human plasma upon incubation at 37 degrees C was shown by the strong inhibitory effect of the anti-PLTP immunoglobulin G. The process of PLTP-mediated HDL enlargement was accompanied by the release of apoproteins, primarily apoA-I. HDL3 enlargement mediated by PLTP was effectively inhibited by the addition of free fatty acids. PMID- 8226828 TI - Real time measurement of nitric oxide produced ex vivo by luminol-H2O2 chemiluminescence method. AB - Endothelium-derived relaxing factor, identified as nitric oxide (NO), is derived from a guanidino nitrogen of L-arginine via its metabolism by nitric oxide synthase. A newly developed aqua-chemiluminescence system was used to measure the amount of NO release from the isolated, perfused rat kidney concomitantly with the pressure changes (Kikuchi, K., Nagano, T., Hayakawa, H., Hirata, Y., and Hirobe, M. (1993) Anal. Chem. 65, 1794-1799). In normotensive rats, basal NO release was estimated to be 86 +/- 6 fmol min-1 (g of kidney weight)-1, and endothelium-dependent vasodilators increased NO release with a concomitant pressure reduction. Pretreatment of the kidney with detergents to damage the vascular endothelium diminished the responses of NO and perfusion pressure to acetylcholine, while infusion of authentic NO solution caused an increase in the chemiluminescence and a concomitant decrease in the pressure. The half-life of NO in O2-saturated Krebs-Henseleit buffer was determined as 6.41 s, and the decay constant (Kd) was 0.108 s-1. The measuring system involves a lag time of 15 s after passage of the solution through the kidney, so that the measured value is 13.0% of the true luminal NO concentration in the kidney, up to 400 pM. These observations enabled us to quantitate the degree to which NO released from the rat kidney changes perfusion pressure. Acetylcholine-induced vasodilation and NO release were markedly attenuated in deoxycorticosterone acetate-salt hypertensive rats, in which the endothelium is known to be markedly damaged. Thus, this assay system may make it possible to evaluate the extent of injury to the endothelium in conditions involving vascular damage. PMID- 8226829 TI - An anion binding site that regulates the glutamate transporter of synaptic vesicles. AB - Glutamate, the major excitatory neurotransmitter of the mammalian central nervous system, is stored in synaptic vesicles and released by exocytosis upon depolarization of the presynaptic nerve terminal. Synaptic vesicles possess an active glutamate-specific transporter that is driven by an electrochemical proton gradient across the vesicle membrane and requires chloride for maximal activity. In this study, we have characterized the role of chloride in vesicular glutamate transport using 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), a potent inhibitor of anion translocators. DIDS inhibited glutamate uptake with an IC50 of 0.7 microM or less. In contrast, all energy gradient parameters (membrane potential, pH gradient, and ATPase activity) required at least 5-fold higher concentration of DIDS for inhibition. Furthermore, high concentrations of chloride but not of glutamate or other anions prevented DIDS inhibition of glutamate uptake. In contrast to uptake, glutamate efflux from glutamate-loaded vesicles was independent of chloride over a wide concentration range. However, efflux was still susceptible to DIDS inhibition. DIDS inhibition was prevented by excess chloride. We conclude that the vesicular glutamate transporter possesses a DIDS-sensitive chloride binding site on the cytoplasmic side, distinct from the substrate binding site, which regulates transport activity. PMID- 8226830 TI - Effect of the relative position of the UGA codon to the unique secondary structure in the fdhF mRNA on its decoding by selenocysteinyl tRNA in Escherichia coli. AB - The fdhF mRNA for formate dehydrogenase H of Escherichia coli contains a UGA codon at position 140. This termination codon is decoded by selenocysteinyl tRNA (the selC product) with the aid of its own specific elongation factor, SelB. For this decoding, a unique secondary structure immediately downstream of the UGA codon has been shown to be essential (Zinoni, F., Heider, J., and Bock, A. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 4660-4664). We examined the positional effect of the UGA codon relative to the secondary structure on its decoding using a fdhF lacZ fusion gene. When the UGA codon was separated by one codon (position -1) from the secondary structure, the UGA decoding, as measured by the beta galactosidase activity, dropped to approximately 76% of the normal level but was still almost as fully dependent upon selC and selenium in the culture medium as in the case of the UGA codon in the normal position (position 0). However, when the UGA codon was separated by two codons (position -2), the decoding level further dropped to 20% of the normal level, and in addition, became dependent only on selC but independent of selenium. When the UGA codon was further separated by three codons (position -3), the decoding level of UGA (-3) became higher than the decoding of UGA (-2) and was completely independent from selC and selenium, indicating that the UGA codon was nonspecifically suppressed. A similar nonspecific suppression was observed for the UGA codon at position -4, but at a lower level. When two UGA codons were tandemly placed at positions 0 and -1, they were still able to be decoded at 17% of the normal level in a selC- and selenium dependent manner. In the absence of the SelB function, the decoding level of UGA(0) dropped to 1.6% of the normal level, whereas the UGA(-1) decoding dropped to 7.5%. These results indicate that the UGA codon at position 0 is not only most effectively decoded by selenocysteinyl tRNA but also tightly blocked from its nonspecific suppression in the absence of any components required for the decoding. PMID- 8226831 TI - Function of the active-site lysine in Escherichia coli serine hydroxymethyltransferase. AB - Serine hydroxymethyltransferase has a conserved lysine residue (Lys-229) that forms the internal aldimine with pyridoxal 5'-phosphate. In other pyridoxal 5' phosphate enzymes investigated so far, this conserved lysine residue also plays a catalytic role as a base that removes the alpha-proton from the amino acid substrate. Three mutant forms of Escherichia coli serine hydroxymethyltransferase (K229Q, K229R, and K229H) were constructed, expressed, and purified. The absorbance spectra, rapid reaction kinetics, and thermal denaturation of the mutant analogs were studied. Only the K229Q mutant serine hydroxymethyltransferase resembled the wild-type enzyme. The results indicate that Lys-229 plays a critical role in expelling the product by converting the external aldimine to an internal aldimine. In the absence of Lys-229, ammonia can also catalyze the same function at a much slower rate. However, Lys-229 apparently is not the base that removes the alpha-proton from the amino acid substrate. The K229Q mutant enzyme could catalyze one turnover of either serine to glycine or glycine to serine at rates approaching those of the wild-type enzyme. After one turnover, the mutant enzyme could not expel the product and bind new substrate. The K229Q mutant enzyme can also transaminate D-alanine, which, like the hydroxymethyltransferase activity, also requires removing the alpha-proton from the substrate. The absorbance spectra of the K229R and K229H serine hydroxymethyltransferases showed that their pyridoxal 5'-phosphate could not readily form an external aldimine with substrates, suggesting that Lys-229 in the wild-type enzyme may never bear a positive charge, further evidence that it is not the base that removes the alpha-proton. PMID- 8226832 TI - Molecular characterization and outer membrane association of a Chlamydia trachomatis protein related to the hsp70 family of proteins. AB - One route by which Chlamydia trachomatis is internalized into host endometrial epithelial cells is receptor-mediated endocytosis. Although this implies an adhesin-receptor interaction exists, specific chlamydial surface molecules have not been identified. We are investigating potential adhesin molecules using an in vitro functional assay to select for chlamydial recombinant Escherichia coli expressing an adherent phenotype. We have previously shown that E. coli JM109(pPBW58) attaches to epithelial cells by a specific process paralleling C. trachomatis and expresses at least three plasmid-encoded proteins (18, 28, and 82 kDa; Schmiel, D. H., Knight, S. T., Raulston, J. E., Choong, J., Davis, C. H., and Wyrick, P. B. (1991) Infect. Immun. 59, 4001-4012). In this report, we demonstrate that (i) the 82-kDa protein is associated with the outer membrane of both E. coli JM109-(pPBW58) and C. trachomatis serovar E elementary bodies; (ii) the plasmid-encoded protein is identical to the native chlamydial protein by mass, charge, antigenicity, and partial proteolytic peptide profiles; (iii) a highly homologous protein is present in C. trachomatis biovariant lymphogranuloma venereum; (iv) the 82-kDa protein is not covalently linked by disulfide bonds to other protein species in either E. coli JM109(pPBW58) or C. trachomatis; (v) sequence analysis of the open reading frame indicates this protein is a relative of the heat shock 70 family of proteins; and (vi) the inferred amino acid sequence contains a contiguous 73-amino acid region having 51% identity with the extracellular sperm receptor binding domain in Strongylocentrosus purpuratus (Foltz, K. R., Partin, J. S., and Lennarz, W. J. (1993) Science 259, 1421-1425). The potential involvement of an hsp70 protein in attachment may provide new insight on adherence mechanisms by obligate intracellular pathogens. PMID- 8226833 TI - Overexpression of fetal human pigment epithelium-derived factor in Escherichia coli. A functionally active neurotrophic factor. AB - Pigment epithelium-derived factor (PEDF) is a neurotrophic protein present in low amounts in conditioned medium of cultured fetal human retinal pigment epithelial cells. Recently, the PEDF cDNA has been cloned from a fetal human cDNA library, and its derived amino acid sequence identified it as a member of the serine protease inhibitor (serpin) supergene family (Steele, F. R., Chader, G. J., Johnson, L. V., and Tombran-Tink, J. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 1526-1530). We have prepared recombinant expression constructs from the fetal human PEDF cDNA and obtained milligram amounts of biologically active PEDF from Escherichia coli. The full-length open reading frame (Met1-Pro418) and a truncated form (Asp44-Pro418) were used in our constructs. Induction from a vector containing the truncated PEDF version, named pEV-BH, produced a protein (BH) of expected size (M(r) 42,800) associated with inclusion bodies, which contained 25-40% of expressed protein. After solubilization, BH was highly purified by gel filtration and cation exchange chromatography. The NH2-terminal sequence of the purified protein matched that of the pEV-BH construct. We have conducted neurite outgrowth assays in a human retinoblastoma Y-79 cell culture system. Recombinant PEDF (BH) demonstrated neurotrophic activity, as reported for the native PEDF. Thus, unfolded and refolded in vitro BH retained a potent biological activity. In parallel experiments, protease inhibition assays were performed. Recombinant PEDF did not have an effect on trypsin, chymotrypsin, elastase, cathepsin G, endoproteinase Lys-C, endoproteinase Glu-C, or subtilisin activity, suggesting that inhibition of known serine proteases is not the biochemical pathway for the PEDF neutrophic activity. PMID- 8226834 TI - Cysteine mutants of light chain-2 form disulfide bonds in skeletal muscle myosin. AB - Multiple disulfide bonds form in recombinant myosin light chain-2 mutants that contain an engineered cysteine at positions 2, 73, or 94, in addition to the endogenous cysteines at residues 126 and 155 (Saraswat, L.D., and Lowey, S. (1991) J. Biol. Chem. 266, 1977). By replacing one of the native cysteines with an alanine, mutants with a single pair of thiols were created: Cys2/Cys155, Cys2/Cys126, Cys73/Cys155 and Cys94/Cys155. Oxidation of these mutants resulted in a fast migrating band on nonreducing SDS gels, which was attributed to an intramolecular disulfide bond. To determine if disulfide formation could also occur when the light chains (LC) are bound to the myosin heavy chains, LCs were added to myosin which had been depleted of its native LC2 by an immunoadsorbent. When the reconstituted myosin was reacted with 5,5'-dithiobis(2-nitrobenzoic acid) in the absence of divalent cations, intramolecular disulfide bonds formed in the mutant and wild type LCs, but the LCs did not remain bound to the myosin heavy chains. Addition of magnesium ions prevented LC dissociation, but intramolecular disulfide bonds no longer formed. Instead, mutants containing cysteines in the NH2-terminal domain formed intermolecular disulfide bonds between the two heads of myosin. The ability to cross-link the heads demonstrates the existence of close head/head interactions in the myosin molecule, a feature that may be essential for regulation. PMID- 8226835 TI - Interaction of non-myristoylated NADH-cytochrome b5 reductase with cytochrome b5 dimyristoylphosphatidylcholine vesicles. AB - An expression vector for NADH-cytochrome b5 reductase containing a thrombin cleavage site directly before the N-terminal glycine residue of the flavoprotein was used to isolate the non-myristoylated enzyme by thrombin cleavage of the initial fusion protein of a short segment of the multiple cloning site of the plasmid vector and the reductase. This flavoprotein preparation, containing only the 28-residue N-terminal peptide segment of the membrane-binding domain of the mammalian enzyme, binds to phospholipid vesicles and interacts with membrane bound cytochrome b5. The effect of N-myristoylation of the enzyme therefore appears to be limited to facilitating and stabilizing interactions with phospholipid vesicles. However, the relatively short intervening peptide sequence that separates the crucial peptide membrane-binding domain from lysine 41, which has been implicated in the active-site interaction with cytochrome b5 (Strittmatter, P., Kittler, J. M., Coghill, J. E., and Ozols, J. (1992) J. Biol. Chem. 267, 2519-2523), provides some limitation of the distance from the membrane surface for the interactions required for rapid electron transfer from the flavin of the reductase to the heme of cytochrome b5. PMID- 8226836 TI - Nucleotide binding sites on beef heart mitochondrial F1-ATPase. Cooperative interactions between sites and specificity of noncatalytic sites. AB - We have studied the properties of beef heart mitochondrial F1 having inhibitory MgADP bound at one of the three catalytic sites and various levels of occupancy of the three noncatalytic nucleotide sites including zero, two, or three ADP/ATPs or two ADP/ATP plus one GTP. The properties examined include the rate of MgATP dependent reactivation and the rate of increase in the fraction of F1 containing transiently bound intermediates. For each form of the enzyme tested, the rate of reactivation closely paralleled the rate of increase in the level of bound intermediates, indicating that when one catalytic site on F1 is blocked by inhibitory MgADP, the remaining two sites are incapable of residual uni- or bi site activity. It was also found that the stability of the MgADP-inhibited complex decreases with full occupancy of the noncatalytic sites. This demonstrates that the noncatalytic sites modulate the properties of catalytic sites. Finally, it was found that the noncatalytic sites on mitochondrial F1 do not, as has long been believed, bind adenine nucleotides exclusively. Evidence is presented that both GTP and PPi bind tightly at noncatalytic sites. PMID- 8226837 TI - A conjugate between human urokinase and saporin, a type-1 ribosome-inactivating protein, is selectively cytotoxic to urokinase receptor-expressing cells. AB - Urokinase-type plasminogen activator (uPA) confers invasive potential to transformed cells. Cancer cells express high numbers of uPA receptors (uPARs), which concentrate uPA activity at the invasive edge of cancer cells and the tumor mass. We synthesized a conjugate between human uPA and saporin (SAP), a ribosome inactivating protein produced by Saponaria officinalis. Results of cell-killing assays showed that uPA is very effective at targeting saporin specifically to uPAR-expressing cells, whereas cell lines devoid of uPARs were not affected by the conjugate. Receptor-bound uPA is internalized only upon formation of a complex with one of its inhibitors (PAIs). However, our conjugate was highly cytotoxic even when the interaction between uPA and PAIs was prevented. Moreover, the alpha 2-macroglobulin receptor, which has been reported to mediate the internalization of uPA.PAI complexes, seems not to be involved in cell killing caused by the uPA.SAP conjugate. Thus, uPA.SAP might follow a mechanism of internalization different from that of unconjugated uPA complexed to PAIs, although still uPAR-mediated. Our results also suggest that alpha 2-macroglobulin and/or its receptor could mediate the internalization and cytotoxicity of unconjugated saporin, as it has been shown for other toxins. PMID- 8226838 TI - Clathrin-coated pit-mediated receptor internalization. Role of internalization signals and receptor mobility. AB - Most signals controlling receptor-mediated endocytosis have been identified by alteration of sequences present in receptors normally internalized via clathrin coated pits. In the present work we have reconsidered the factors that control internalization the other way around: i.e. by introducing potential internalization sequences in complement receptor 1 (CR1) which does not preferentially associate with clathrin-coated pits. The analysis of the internalization efficiency of NPxY related motifs generated by substituting His2010 and/or Glu2015 by either Phe or Tyr indicates that FxNPxY is the stronger promoter of endocytosis and that the signal efficiency depends on the presence of aromatic residues (including a tyrosine) at both ends of the -xNPx- motif. Moreover, CR1-tyr (substitution of Glu2015 for Tyr) internalization was superposable to that of a receptor composed of the extracellular and transmembrane domains of CR1 fused to the intracytoplasmic tail of the low density lipoprotein (LDL) receptor (including the FxNPxY motif) (CR1-LDL). When analyzed by fluorescence recovery after photobleaching, the surface mobility of CR1-LDL was decreased as compared with that of either CR1-tyr or CR1-wt, despite a similar association with clathrin-coated pits. The role of receptor mobility in internalization was confirmed by the observation that CR1-tl, with a deletion of the cytoplasmic tail, was more mobile and more efficiently internalized than CR1 wt. PMID- 8226839 TI - Deconjugation of bilirubin-IX alpha glucuronides: a physiologic role of hepatic microsomal beta-glucuronidase. AB - beta-Glucuronidase is an acid hydrolase located in both the lysosomal and microsomal compartments of the hepatocyte. The function of the latter remains undefined. We postulated that microsomal beta-glucuronidase may be responsible for the deconjugation of bilirubin-IX alpha glucuronides which are synthesized primarily in the hepatic microsomal compartment. We utilized two unique congenic strains of mice to characterize the role of hepatic beta-glucuronidase in the metabolism and disposition of bilirubin-IX alpha; the first exhibited less than 1% of total hepatic beta-glucuronidase activity (ATM), the second lacked only the microsomal enzyme activity (AT1). The biliary excretion of bilirubin-IX alpha conjugates was quantitated using reverse-phase high performance liquid chromatography. Under basal conditions, there was a 2-fold increase in the biliary excretion of bilirubin-IX alpha monoglucuronides and total glucuronides in the AT1 and ATM mutants compared to the normal controls. When the plasma bilirubin-IX alpha level was increased to approximately 7 mg/dl to simulate hyperbilirubinemia, by intravenous administration of [14C]bilirubin-IX alpha, mathematical modeling of the biliary excretion curves of bilirubin-IX alpha glucuronides revealed qualitative differences between control and mutant animals, whereas both mutant groups were similar. Collectively, these data demonstrate that microsomal beta-glucuronidase modulates the net rate of bilirubin-IX alpha glucuronidation and glucuronide excretion in bile, under both basal and hyperbilirubinemic conditions, and that lysosomal beta-glucuronidase has no such effects. Hepatic microsomal beta-glucuronidase appears likely to influence the biliary excretion and hence the hepatic elimination of endogenous and xenobiotic substrates (e.g. carcinogens) which undergo hepatic glucuronidation. PMID- 8226840 TI - The carboxyl terminus of nerve growth factor is required for biological activity. AB - A series of mouse nerve growth factor (NGF) deletion mutants have been constructed using in vitro mutagenesis to define domains of the protein essential for its activity. Deletions of the amino or carboxyl termini of mature NGF or of an internal domain, which normally produces a surface-exposed reverse turn, have been analyzed. Mutants with deletions in the amino terminus or in the reverse turn retain significant biological activity, whereas, in contrast, a mutant NGF lacking the seven most carboxyl-terminal amino acids is appropriately synthesized but shows no measurable biological activity. These results suggests that the flexible carboxyl tail of NGF, and perhaps other neurotrophins, plays a crucial role in mediating receptor recognition and/or ligand binding. PMID- 8226841 TI - The multiphosphorylation domain of the phosphorylase kinase alpha M and alpha L subunits is a hotspot of differential mRNA processing and of molecular evolution. AB - We have cloned and sequenced human cDNAs encoding the complete phosphorylase kinase alpha subunit muscle isoform (alpha M). The predicted polypeptide is highly similar to the sequence known from rabbit muscle but lacks a major part of its multiphosphorylation domain, including the main phosphorylation site for cAMP dependent protein kinase (PKA). Analysis of this region by reverse-transcribed polymerase chain reaction (RT-PCR) in several human and rabbit tissues demonstrates that it is subject to elaborate differential mRNA splicing. Amino acids 1012-1024 of the full-length rabbit sequence, including the major PKA phosphorylation site, and amino acids 1025-1041, which harbor at least one endogenous phosphorylation site, can be deleted from the predicted polypeptide individually or in combination. Molecules lacking one or both of these segments constitute a major part of the alpha M subunit population in many rabbit tissues and constitute the vast majority in all human tissues analyzed. Similar, tissue dependent differential splicing events could be detected by RT-PCR in the human alpha subunit isoform from liver (alpha L). The expression of the differentially spliced alpha M subtypes differs markedly between corresponding human and rabbit tissues. Sequence divergence in this region is particularly high, not only between the muscle and liver isoforms, but also between alpha M sequences from four different animal species. Moreover, a duplication of the exon encoding the main PKA phosphorylation site was discovered in the mouse. Thus, the multiphosphorylation domain of the phosphorylase kinase alpha subunit isoforms is subject to pronounced structural variation not only between different tissues of one organism via differential splicing, but also in the course of evolution. PMID- 8226842 TI - NAD-binding site of the C3-like ADP-ribosyltransferase from Clostridium limosum. AB - Treatment of the Rho-ADP-ribosylating C3-like transferase from Clostridium limosum by ultraviolet irradiation in the presence of [carbonyl-14C]NAD incorporated 1 mol of label/mol of exoenzyme. Concomitantly, the transferase and NAD glycohydrolase activity was impaired. A peptide containing the radiolabel was obtained by proteolysis with either staphylococcal protease V8 or trypsin. Their amino acid sequences were Ala/Asp-Gly-Tyr-Ile-Glu-Pro-Ile-Ser-Thr-Phe-Lys-Gly-Gln Leu-X-Val-Leu-Le u-Pro- Arg and Gly-Gln-Leu-X-Val-Leu-Leu-Pro-Arg, respectively. These sequences correspond with regions Ala-160 through Arg-179 and Gly-171 through Arg-179, respectively, of the very similar Clostridium botulinum C3 transferase, with X being Glu in the unlabeled enzyme. This identifies the glutamic acid residue that corresponds to Glu-174 of C. botulinum C3 transferase as part of the NAD-binding site of the catalytic center of the C. limosum exoenzyme. PMID- 8226843 TI - Calcium oscillations in pancreatic acinar cells, evoked by the cholecystokinin analogue JMV-180, depend on functional inositol 1,4,5-trisphosphate receptors. AB - It has been reported that the synthetic heptapeptide cholecystokinin (CCK) analogue JMV-180 evokes cytosolic Ca2+ signals in pancreatic acinar cells via mechanisms that do not include either the generation or action of inositol 1,4,5 trisphosphate (InsP3) (Saluja, A. K., Dawra, R. K., Lerch, M. M., and Steer, M. L. (1992) J. Biol. Chem. 267, 11202-11207; Yule, D. I., and Williams, J. A. (1992) J. Biol. Chem. 267, 13830-13835). We have now investigated the CCK- and JMV-180-evoked cytosolic Ca2+ oscillations by measurement of the Ca(2+)-sensitive ion currents in internally perfused mouse pancreatic acinar cells. We find that the InsP3 receptor antagonist heparin (500 micrograms/ml) blocks Ca2+ oscillations induced by both CCK (5-20 pM) and JMV 180 (10-40 nM), whereas de-N sulfated heparin (500 micrograms/ml), which does not affect InsP3 binding to its receptor, fails to inhibit the responses to the two agonists. We conclude that the cytosolic Ca2+ oscillations evoked by both CCK and JMV-180 are dependent on functional InsP3 receptors. PMID- 8226844 TI - Riboflavinuria in the rd chicken. 5'-splice site mutation in the gene for riboflavin-binding protein. AB - Eggs of oviparous species must contain all components required for normal embryonic development. Among these, the vitamin riboflavin is deposited into egg white and yolk bound to the carrier protein, riboflavin-binding protein (ribBP). We have begun to investigate the mechanisms underlying ribBP transport pathways by molecular characterization of a relevant mutation in chicken. The autosomal recessive rd allele in Gallus gallus domesticus prevents the synthesis of functional ribBP and induces embryonic death on day 13. Polymerase chain reaction primers derived from the previously determined wild type cDNA sequence were used to amplify and clone cDNAs for ribBP from normal (Rd) and deficient (rd) animals. The rd allele was associated with a 100-nucleotide deletion in the messenger RNA for ribBP. Genomic clones were generated via polymerase chain reaction using primers flanking this 100-base pair deletion, and the resulting 2.1-kilobase pair clones were sequenced. The deletion in the rd ribBP cDNA corresponds precisely to an exon. The splice site following this exon contains a G-->A mutation at position 1 of the downstream 5'-splice donor sequence. The effect of this anomaly and the cause of the rd phenotype is the loss of the 100-base pair exon during the splicing process. PMID- 8226845 TI - An estrogen receptor mutant exhibiting hormone-independent transactivation and enhanced affinity for the estrogen response element. AB - To study transactivation by the Xenopus laevis estrogen receptor (XER), we inserted one or two copies of a synthetic amphipathic helix at amino acid 276 of the XER. The XER mutants containing one or two copies of the amphipathic helix (XER/1AH and XER/2AH, respectively) and wild-type XER were expressed at similar levels. In transient transfection assays, XER/1AH exhibited only a modest, promoter-specific increase in transactivation. Constitutive (estrogen independent) transcription of a synthetic promoter containing two estrogen response elements (EREs) was approximately 10-fold higher for the XER/2AH mutant than for wild-type XER. The XER/2AH mutant and wild-type XER exhibited similar 17 beta-estradiol dose-response curves for transactivation. In studies carried out over a broad range of DNA concentrations using the simple 2ERE-TATA promoter or a complex vitellogenin-derived promoter, the XER/2AH mutant exhibited an estrogen dependent 2-3-fold increase in transactivation. A 2-3-fold increase in transactivation by XER/2AH was also observed using synthetic promoters in which the two EREs exhibit synergistic interactions with the NF1, AP1, or vitellogenin activator upstream activator sequences. Using a promoter interference assay to investigate intracellular interactions between the estrogen receptor and the ERE, we showed that binding of wild-type XER to the ERE was strongly estrogen dependent. In the presence of 17 beta-estradiol, XER/2AH and wild-type XER exhibited similar promoter interference curves. In the absence of 17 beta estradiol, the expression plasmid encoding the XER/2AH mutant achieved levels of promoter interference with 0.25-0.5 microgram of transfected DNA that were similar to those observed with 5-10 micrograms of the expression plasmid encoding wild-type XER. The ability of the XER/2AH mutant to activate transcription in the absence of estrogen therefore is likely to be related to the approximately 20 fold increase in its apparent ability to bind to the ERE. Since XER/2AH was unable to activate transcription from a glucocorticoid response element, enhanced binding of XER/2AH to the ERE did not result from a general increase in binding to DNA. The XER/2AH mutant appears to be the first nuclear receptor mutant to retain hormone-dependent transactivation and to exhibit enhanced hormone independent binding to its hormone response element. PMID- 8226846 TI - Dependence of trp repressor-operator affinity, stoichiometry, and apparent cooperativity on DNA sequence and size. AB - A series of chemically synthesized trp and mutant operator DNAs was employed to examine trp repressor binding. Although only a single repressor-operator complex was observed for most DNAs as reported previously, varying DNA sequence revealed two retarded complexes with an additional band of faster mobility. The relative intensity of the two retarded bands with varying repressor concentrations suggests that cooperative interactions between dimers may occur in the formation of the predominant repressor-operator complex. Direct stoichiometry measurements demonstrated that a 2:1 stoichiometry (two dimers per operator) is found in the primary repressor-operator complex band and that a 1:1 stoichiometry is observed, when present, for the minor repressor-operator complex band of faster mobility. Similar retardation patterns with a single complex of 2:1 stoichiometry were observed for 40-base pair (bp) trp operators corresponding to TrpEDCBA, aroH, and Trp-PL (a derivative of TrpEDCBA with increased symmetry) operator sequences as well as to hybrid operators containing a half-binding site from TrpEDCBA in conjunction with lac operator sequences, although the apparent affinity for the half-site DNAs was diminished by 10-fold. In contrast, the prominence of the 1:1 dimer-operator complex for Trp-PR, a different derivative of TrpEDCBA with increased symmetry, suggests that sequence context may diminish cooperativity between dimers. The stoichiometry observed was also dependent on the length of TrpEDCBA operator DNA used, shifting from primarily 2:1 for 40- and 33-bp TrpEDCBA DNA to primarily 1:1 for 29-, 26-, and 20-bp TrpEDCBA DNAs. In addition, the stability of the repressor-operator complex to electrophoresis is reduced for DNA lengths of 33, 29, and 26 bp. Based on the binding data and footprinting patterns for two hybrid 40-bp TrpEDCBA/lac half-binding site DNAs, it appears that repressor associates tightly to the specific trp half-site, whereas the nonspecific half of the DNA is more loosely bound. These results suggest that repressor dimer-dimer interaction may be an important feature in the trp repressor-operator interaction. PMID- 8226847 TI - Effect of cholesterol on the charge and structure of apolipoprotein A-I in recombinant high density lipoprotein particles. AB - The effects of cholesterol on the conformation and net charge of apoA-I have been investigated in homogeneous recombinant high density lipoprotein (HDL) particles. ApoA-I charge and structure in discoidal recombinant HDL complexes containing palmitoyloleoylphosphatidylcholine and cholesterol have been quantitated by guanidine HCl denaturation, circular dichroism, electrokinetic analysis, and NMR spectroscopy of [13C]lysine-labeled apoA-I. In a discoidal particle containing 2 molecules of apoA-I and 160 molecules of palmitoyloleoylphosphatidylcholine, apoA I exhibits an alpha-helix content of 75%, and the particle has a net negative surface charge of -5.2e/mol of apoA-I at pH 8.6. Addition of 2 molecules of cholesterol to this complex has no significant effect upon particle size, but slightly decreases the net charge (-5.0e) and alpha-helix content (68%) of apoA-I and enhances the stability of the helical segments, as reflected by an increase in the free energy of unfolding from 2.9 to 3.5 kcal/mol. In contrast, increasing the cholesterol content to 20 molecules/particle progressively increases particle size and apoA-I net negative charge (-6.1e), and there is a concomitant reduction in the free energy of stabilization of the alpha-helical structure in apoA-I to 2.2 kcal/mol. (13CH3)2-Lys resonances from apoA-I in discoidal recombinant HDL exhibit six chemical shifts at pH 10; these peaks originate from dimethyl-Lys residues that have pKa values ranging from 8.4 to 10.3. The titration behavior of apoA-I Lys residues is generally similar in the presence and absence of cholesterol, except that 4 Lys residues titrate at a significantly higher pH in the presence of cholesterol. These data are consistent with cholesterol having a direct effect on apoA-I conformation and charge in HDL. Structural changes of this magnitude can affect the interactions between HDL and various plasma proteins and cell surfaces. It is therefore likely that the cholesterol content of HDL plays an important role in regulating the metabolism of this lipoprotein. PMID- 8226848 TI - Transcripts for the high and low affinity hexose transporters in rat myoblasts. AB - The present investigation examined the glucose transporter (GLUT) transcript levels in glucose-grown rat L6 myoblasts, which possessed a high (HAHT) and a low (LAHT) affinity hexose transport system. The most prominent GLUT transcript detected in these cells was a GLUT 3-recognizable 4.1-kilobase transcript. Mutants defective in HAHT contained only residual levels of the GLUT 3 transcript and transporter. Corresponding changes in GLUT 3 transcript levels and HAHT activity were observed in myotubes, confluent cultures, and myogenesis-impaired cells. The other prominent GLUT transcripts detected were the GLUT 4-recognizable 2.8- and 1.4-kilobase transcripts. The close correlation between the GLUT 4 isoform and LAHT activity was indicated by the absence of this isoform in LAHT- mutants, and their corresponding changes in myoblasts grown under different conditions. These findings suggested the HAHT and LAHT transporters might be coded or regulated by the genes responsible for the GLUT 3 and 4 transcripts, respectively. Although relatively high levels of the GLUT 1 transcript and transporter were detected in glucose-grown HAHT-LAHT- mutants, these cells did not exhibit any GLUT 1 transport activity. Since the GLUT 1 transporter was not functional, it was not likely responsible for the HAHT and LAHT activities observed in glucose-grown L6 myoblasts. PMID- 8226849 TI - Aginactin, an agonist-regulated F-actin capping activity is associated with an Hsc70 in Dictyostelium. AB - We have previously isolated an agonist-regulated actin filament capping activity, called aginactin, that is associated with a 70-kDa protein (Sauterer, R. A., Eddy, R. J., Hall, A. L., and Coneelis, J. S. (1991) J. Biol. Chem. 266, 24533 24539). A 2.0-kilobase clone isolated from a Dictyostelium lambda gt11 cDNA library screened with affinity-purified aginactin antibodies displays an overall sequence identity of 73% to the 70-kDa heat shock cognate protein, Hsc70, from various species. Aginactin capping activity and the 70-kDa protein bind to ATP agarose columns and are quantitatively depleted from the load, indicating that an Hsc70 is associated with aginactin activity. Moderate stringency Southern blots indicate the presence of no fewer than six Hsc70-related sequences. Immunofluorescent staining of vegetative Dictyostelium AX3 cells with aginactin antibodies reveals a colocalization of aginactin-associated Hsc70 in F-actin-rich regions of the cell cortex and cell protrusions. Nuclei and organelles lacked positive staining indicating that the aginactin-associated Hsc70 is cytosolic. The levels of cytoskeletal-associated Hsc70 correlate with the loss of barbed end capping activity following cAMP stimulation, suggesting that the uncapping of barbed filament ends through an Hsc70-associated process may account for the increase in nucleation activity observed at 5 s following agonist stimulation. PMID- 8226850 TI - Phosphorylation of serine 59 of p56lck in activated T cells. AB - p56lck, a member of the src family of non-receptor protein tyrosine kinases, is expressed almost exclusively in cells of lymphoid origin. Recent evidence has implicated p56lck in a critical role both in T cell development and activation. A variety of T cell stimuli induce a shift in the electrophoretic mobility of p56lck from an apparent molecular mass of 56 kDa (p56lck) to 60 kDa (p60lck). This shift in electrophoretic mobility correlates with an increase in the phosphoserine content of the p60lck. We have shown that both 4 alpha-phorbol 12 beta-myristate acetate and OKT3 treatment of Jurkat cells, as well as 4 alpha phorbol 12 beta-myristate acetate treatment of 171.CD4 and LSTRA cells, induced phosphorylation of serine-59 on p56lck in vivo, which correlated with the shift to p60lck. We also demonstrated that the same serine residue could be phosphorylated in vitro with mitogen-activated protein kinases and that this event was capable of reducing p56lck activity in vitro. Combined, these data suggest a novel pathway for the in vivo regulation of p56lck activity. PMID- 8226851 TI - The effect of retinoic acid on amino acid uptake and protein synthesis by lung fibroblasts. AB - The effect of retinoic acid (RA) on the uptake and utilization of extracellular amino acids by fetal lung fibroblasts was examined. RA decreased the incorporation of [3H]proline into collagen and other proteins. The effect was maximal at a RA concentration of 10(-5) M; smaller decreases were observed at a RA concentration of 10(-6) M. This decrease in collagen formation was associated with a large decrease in intracellular [3H] proline. The decrease in intracellular [3H]proline was first observed at 2 h following the addition of RA to cell cultures. Transport studies employing radiolabeled amino acids revealed that RA decreased the uptake of proline, 2-aminoisobutyric acid, and 2 (methylamino)isobutyric acid but not leucine or methionine. Kinetic analysis of 2 aminoisobutyric acid uptake indicated that this effect was mediated primarily by an increase in apparent Km, with a lesser decrease in Vmax, RA-induced inhibition of proline uptake was not abolished by the presence of cycloheximide nor by pretreatment with indomethacin. Na+,K(+)-ATPase activity was not affected by RA treatment. These results suggest that RA modulates protein production in fibroblasts by altering the function of the Na(+)-dependent A transport system for amino acid uptake. PMID- 8226852 TI - Mitochondrial voltage-dependent anion channel. Immunochemical and immunohistochemical characterization in rat brain. AB - The purified mitochondrial benzodiazepine receptor (mBzR) is a complex comprising the voltage-dependent anion channel (VDAC), adenine nucleotide carrier, and an 18 kDa protein that binds isoquinoline carboxamide ligands (McEnery, M. W., Snowman, A. M., Trifiletti, R. R., and Snyder, S. H. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 3170-3174). An antiserum raised against the mBzR complex reacts selectively with VDAC and is used, along with purification, electrophysiological and immunohistochemical techniques, to characterize the properties and distribution of rat brain VDAC. Although purified VDAC displays biochemical and electrical conductance properties similar to VDAC from other sources, the immunohistochemical distribution of VDAC in rat brain is heterogeneous with pronounced regional variations; the pontine nuclei, the supraoptic nucleus, Purkinje cells of the cerebellum, and the caudate putamen evidence the highest density. The distribution of VDAC is inclusive of the more discretely localized 18-kDa mBzR protein, suggesting that only a portion of the total VDAC participates in the mBzR. The histochemical localizations of the mitochondrial marker enzymes glutamate dehydrogenase and cytochrome c oxidase also indicate marked regional variability in both mitochondrial content and composition. The discrete expression of VDAC reflects a striking heterogeneity of rat brain mitochondria and underlying differences in the utilization of mitochondrial outer membrane ion channels. PMID- 8226853 TI - Mastoparan stimulates exocytosis at a Ca(2+)-independent late site in stimulus secretion coupling. Studies with the RINm5F beta-cell line. AB - Mastoparan, a tetradecapeptide from wasp venom, stimulated exocytosis in a concentration-dependent manner, which was enhanced by pertussis toxin pre treatment, in the insulin secreting beta-cell line RINm5F. Mastoparan (3-20 microM) also elevated cytosolic free calcium concentration ([Ca2+]i), a rise that was not attenuated by nitrendipine. Divalent cation-free Krebs-Ringer bicarbonate (KRB) medium with 0.1 mM EGTA nullified the mastoparan-induced increase in [Ca2+]i, suggesting that the peptide increased Ca2+ influx but not through the L type voltage-dependent Ca2+ channel. Depletion of the intracellular Ca2+ pool did not affect the mastoparan-induced elevation of [Ca2+]i. Remarkably, in divalent cation-free KRB medium with 0.1 mM EGTA and 2 microM thapsigargin in which mastoparan reduced [Ca2+]i, the mastoparan-stimulated insulin release was similar to that in normal Ca(2+)-containing KRB medium. Inhibitors of protein kinase C, such as bisindolylmaleimide, staurosporine, and 1-O-hexadecyl-2-O-methyl-rac glycerol did not suppress the mastoparan-stimulated insulin release. Mastoparan at 10-20 microM did not increase cellular cAMP levels, nor did mastoparan at 5-10 microM affect [3H]arachidonic acid release. In conclusion, although mastoparan increased [Ca2+]i, this increase was not involved in the stimulation of insulin release. Rather, the data suggest that mastoparan directly stimulates exocytosis in a Ca(2+)-independent manner. As GTP-binding proteins (G proteins) are thought to be involved in the process of exocytosis and as mastoparan is known to exert at least some of its effects by activation of G proteins, an action of mastoparan to activate the putative stimulatory Ge (exocytosis) protein is likely. PMID- 8226854 TI - Formation of a stable inactive complex of the sarcoplasmic reticulum calcium ATPase with magnesium, beryllium, and fluoride. AB - Incubation of leaky or nonionic detergent-solubilized sarcoplasmic reticulum vesicles in solutions containing magnesium, beryllium, and fluoride caused a time dependent complete inhibition of calcium ATPase activity. The inhibited state persisted through dialysis for 2 days versus EGTA and was reversed within minutes by the presence of 0.5 mM calcium. Calcium-independent ATPase activity was unaffected. Omission of magnesium or fluoride resulted in retention of activity, while omission of beryllium produced slower inactivation, as described previously (Murphy, A. J., and Coll, R. J. (1992) J. Biol. Chem. 267, 5229-5235). Incubation of nonleaky vesicles had a similar effect, although it occurred more than 10-fold more slowly, suggesting that a component, probably beryllium, must enter the vesicles for inhibition to occur. By contrast, inhibition of nonleaky vesicles by magnesium and fluoride developed less than 2-fold more slowly. Including calcium in the incubation mixtures resulted in partial protection, so that the time course of CaATPase activity leveled off at nonzero values (for example, at 0.1 mM calcium, the activity leveled off at 43% of control). This result is most simply accounted for by a model involving simultaneous binding of calcium and a form of fluoroberyllium to the CaATPase (e.g. Ca2EMgBeFn). PMID- 8226855 TI - Cloning and functional expression of a cDNA encoding the catalytic subunit of bovine enterokinase. AB - Enterokinase (enteropeptidase) is a heterodimeric serine protease that is responsible for the physiological activation of trypsinogen by highly specific cleavage of the trypsinogen activation peptide following the sequence (Asp)4-Lys. In this paper, we report the cloning and functional expression of a cDNA encoding the catalytic domain (light chain) of bovine enterokinase. The nucleotide sequence of this cloned cDNA predicts a 235-amino acid polypeptide that shares a high degree of homology with a variety of mammalian serine proteases involved in digestion, coagulation, and fibrinolysis. We have developed a novel expression method for the enzyme which utilizes the secretory leader and propeptide of the mammalian serine protease PACE fused to the enterokinase light chain amino terminus. Efficient cleavage of the paired dibasic amino acid cleaving enzyme (PACE) propeptide was achieved by coexpression with human PACE or yeast KEX2. The mature product migrates at 43,000 Da on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, comparable to light chain derived from bovine duodena, and exhibited high levels of activity in cleaving the enterokinase-specific fluorogenic substrate Gly-(Asp)4-Lys-beta-naphthylamide. The recombinant single chain form of enterokinase was also capable of activating trypsinogen, indicating that the specificity of the enzyme for its natural substrate is retained even in the absence of the noncatalytic enterokinase heavy chain. PMID- 8226856 TI - Sulfite both stimulates and inhibits the yeast vacuolar H(+)-ATPase. AB - The yeast vacuolar H(+)-ATPase (V-ATPase) exhibits nonlinear hydrolysis kinetics, i.e. an initial rapid rate followed by a slower, steadily declining rate. Sulfite (50-100 mM) stimulates the yeast V-ATPase specifically during the latter period. Sulfite activation has been observed for the F-ATPases and archaebacterial ATPases and is thought to be caused by the release of tightly bound nucleotide at the catalytic site. However, turnover-dependent inactivation of the yeast V ATPase, and sulfite stimulation, were only observed at MgATP concentrations > 1.0 mM. Below 1.0 mM MgATP, the hydrolysis time course was linear, and sulfite was inhibitory. The inhibition during the initial phase and the stimulation during the later phase of the time course could be accounted for by a 5.5-fold sulfite induced increase in the apparent Km, and a small increase in the apparent Vmax. Sulfite also protected the enzyme against inhibition by cold inactivation and by dicyclohexylcarbodiimide but not by bafilomycin. Sulfite stimulation during the later phase was antagonized by delta mu H+, particularly by delta pH. In contrast to its effects on hydrolysis, sulfite inhibited the formation of a pH gradient at all times and failed to enhance the membrane potential even when delta pH was collapsed by nigericin. These results indicate that sulfite partially uncouples hydrolysis from proton transport in a way that preserves regulation by delta mu H+. PMID- 8226857 TI - Activation of protein-tyrosine kinase p72syk with concanavalin A in polymorphonuclear neutrophils. AB - We studied the abundance, subcellular distribution of a non-receptor protein tyrosine kinase p72syk (Taniguchi, T., Kobayashi, T., Kondo, J., Takahashi, K., Nakamura, H., Suzuki, J., Nagai, K., Yamada, T., Nakamura, S., and Yamamura, H. (1991) J. Biol. Chem. 266, 15790-15796) in porcine polymorphonuclear neutrophils and the activation upon the stimulation with concanavalin A. The abundance was about 0.1% of total proteins and mainly distributed in the particulate fraction. Upon concanavalin A stimulation, the activity of p72syk increased within 30 s, attained to the maximum level at 1 min, and then returned to the basal level within 6 min. This activation was observed in a dose-dependent manner and abrogated by simultaneous addition of methyl alpha-mannopyranoside. When both extra- and intracellular Ca2+ were depleted, the activation of p72syk was still persistent; in contrast, the deactivation process was completely abrogated even at 6 min after stimulation. The replenishment of Ca2+ in the presence of A23187 resulted in a similar deactivation pattern as seen in the Ca(2+)-rich condition. In addition, genistein and herbimycin A, potent protein-tyrosine-kinase inhibitors, were capable of reducing concanavalin A-evoked p72syk activation and Ca2+ mobilization as well as the aggregation and lysozyme release. Furthermore, A23187-induced Ca2+ accumulation in inhibitor-treated cells resulted in the restoration of those cellular responses. These lines of evidence suggest that p72syk is activated with concanavalin A in a Ca(2+)-independent manner, participating in a mechanism of Ca2+ recruitment, and negatively regulated by a feedback mechanism through Ca2+ in neutrophils. PMID- 8226858 TI - The effect of aspartate hydroxylation on calcium binding to epidermal growth factor-like modules in coagulation factors IX and X. AB - Hydroxylation of aspartic acid to erythro-beta-aspartic acid (Hya) occurs in epidermal growth factor (EGF)-like modules in numerous extracellular proteins with diverse functions. Several EGF-like modules with the consensus sequence for hydroxylation bind Ca2+, and it has therefore been suggested that the hydroxyl group is essential for Ca2+ binding. To determine directly the influence of beta hydroxylation on calcium binding in the EGF-like modules from coagulation factors IX and X, we have now measured calcium binding to both the fully beta hydroxylated and the non-beta-hydroxylated modules of the two proteins. At low ionic strength, the Hya-containing module of factor X has a slightly higher Ca2+ affinity, but at physiological salt concentrations this difference is no longer significant for either factor IX or X. Analysis of the 1H NMR chemical shift differences between the hydroxylated and nonhydroxylated factor X modules show that hydroxylation has no effect on the domain fold. Furthermore, measurements on factor IX show that hydroxylation has no effect on the Ca2+/Mg2+ specificity of the ion binding site. We conclude that the hydroxyl group is not a direct ligand for the calcium ion in these EGF-like modules, nor is it essential for high affinity Ca2+ binding. PMID- 8226859 TI - The heat stable antigen (mouse CD24) gene is differentially regulated but has a housekeeping promoter. AB - Expression of the mouse heat stable antigen (HSA or mouse CD24) shows tissue specific as well as developmental regulation. During the maturation of several hematopoietic lineages, HSA expression is generally high in immature precursor cells and low or absent in terminally differentiated cells. We present evidence suggesting that this regulation of the HSA gene (Cd24a) occurs at the transcriptional level. In addition, sequence and methylation analysis of the Cd24a promoter revealed characteristics of both "housekeeping" and tissue specific promoters, including a methylation-free, HpaII tiny fragment (HTF) island, multiple putative SP1 and AP-2 consensus binding sites, and a TATA box. Functional analysis of a 0.6-kilobase DNA fragment containing these elements fused to the CAT reporter gene in transient transfection experiments showed activity in both HSA expressing and non-expressing cell lines with a strength similar to that of the herpes-simplex virus-thymidine kinase promoter. Large fragments from the flanking region of the Cd24a promoter did not influence the ubiquitous nature of this promoter. Finally, we mapped the Cd24a, Cd24b, and Cd24c genes to mouse chromosomes 10, 8, and 14 respectively. PMID- 8226860 TI - Isolation and structural characterization of monomeric and trimeric photosystem I complexes (P700.FA/FB and P700.FX) from the cyanobacterium Synechocystis PCC 6803. AB - An isolation procedure was developed for the cyanobacterium Synechocystis 6803 (and 6714) which yields both monomeric and trimeric photosystem I complexes (P700.FA/FB complexes) depleted of the stroma-exposed subunits PsaC, -D, and -E (P700.FX complexes). Analysis by high resolution gel electrophoresis in combination with immunoblotting and N-terminal sequencing reveals the selective and quantitative removal of PsaC, -D, and -E from the P700.FA/FB complex, containing PsaA, -B, -C, -D, -E, -F, -K, -L and at least two subunits < or = 4 kDa. Monomeric and trimeric P700.FX complexes show an identical subunit composition and an identical charge recombination half-time of 750 +/- 250 microseconds as determined by flash-induced absorption change measurements, reflecting the quantitative loss of iron-sulfur clusters FA/FB and the presence of cluster FX. The existence of a stable trimeric P700.FX complex enables a detailed structural analysis by electron microscopy with high resolution. Comparison of averaged top and side view projections of P700.FX and P700.FA/FB complexes show that the height of the complex is reduced by about 2.5-3.3 nm upon removal of the three stroma-exposed subunits and indicate the position of these three subunits on the PS I surface. While the outer contours of the stroma exposed mass of PS I agree very well with the three-dimensional crystal analysis recently published for trimeric PS I of Synechococcus elongatus (Krauss, N., Hinrichs, W., Witt, I., Fromme, P., Pritzkow, W., Dauter, Z., Betzel, C., Wilson, K. S., Witt, H. T., and Saenger, W. (1993) Nature 361, 326-330), only the structural analysis presented here is able to assign the stroma-exposed mass exclusively to the subunits PsaC, -D, and -E and to exclude a contribution of other subunits. PMID- 8226861 TI - An AP-1-like factor and the pituitary-specific factor Pit-1 are both necessary to mediate hormonal induction of human thyrotropin beta gene expression. AB - The human thyrotropin beta (hTSH beta) gene is inducible by various agents including thyrotropin-releasing hormone, phorbol esters, or the adenylyl cyclase activator forskolin. In this study, we have characterized the functional properties of the TGGGTCA motif at -1/+6 of the hTSH beta gene that is similar to the consensus phorbol ester response element (TRE) or the consensus cyclic AMP response element (CRE). We suggest that both protein kinases C and A as well as TRH share a common mediator which recognizes the TGGGTCA element in activating the hTSH beta promoter. Following stimulation by phorbol esters, forskolin, or TRH, the TGGGTCA-specific factor acts together with the pituitary-specific transcription factor Pit-1 (or GHF-1) bound to upstream sequences at -128 to -61 to mediate the induction of the hTSH beta promoter. The induction requires that both factors bind to their own binding sites, but Pit-1 neither increases the binding of the TGGGTCA-specific factor to its target sequences nor associates with this factor to form a heterodimer. The TGGGTCA-specific factor is present in three cell lines tested and is composed of protein(s) immunologically related to c-Jun and c-Fos but not to the CRE-binding protein, CREB. By using the hTSH beta reporter plasmids in which the TGGGTCA element is converted to consensus TRE or CRE motifs, we found that, within the context of the hTSH beta promoter, the TGGGTCA element is a more potent TRE or CRE than the consensus TRE or CRE sequences. Based upon the results of this study, we propose a model in which the TGGGTCA-specific AP-1-like factor functionally cooperates with the tissue specific factor Pit-1 to activate the hTSH beta gene. PMID- 8226862 TI - Sequence and regulation of the uapA gene encoding a uric acid-xanthine permease in the fungus Aspergillus nidulans. AB - The nucleotide sequence of the uapA gene, coding for the uric acid-xanthine permease of Aspergillus nidulans, has been determined. The predicted uapA gene product comprises 595 amino acids (M(r) 63,365); it is a highly hydrophobic protein with 12-14 putative transmembrane segments and shows no striking similarity to any other membrane protein of either prokaryotes or eukaryotes, except for a short highly hydrophobic amino acid sequence conserved in a number of different permeases. The presence of an acidic, amphipathic region overlapping with the last hydrophobic segment of UAPA could also be of interest. The results presented suggest that the UAPA permease represents a new type of membrane protein, not described previously. The transcription of uapA is inducible by 2 thiouric acid, and it is highly repressible by ammonium. It is almost absolutely dependent on the presence of functional uaY and areA regulatory gene products. A specific mutation in the GATA binding zinc finger of the AREA protein nearly abolishes uapA transcription. The uap100 cis-acting, up-promoter, constitutive mutation is a duplication that comprises two GATA sites and suppresses weakly the AREA zinc finger mutation but does not alleviate the need for functional UAY and AREA proteins. PMID- 8226863 TI - Sequence, regulation, and mutational analysis of the gene encoding urate oxidase in Aspergillus nidulans. AB - The uaZ gene of Aspergillus nidulans codes for a protein of 301 amino acids. The open reading frame is interrupted by two introns. A comparison with the open reading frames of 10 other urate oxidases reveals a number of scattered universally conserved residues and four clusters of high similarity. Two of these are of unknown function, and the other two are the putative sites for copper binding and the signal sequence for peroxisomal entry, respectively. This comparison also reveals that there are at 16 positions at which introns can be found in the gene coding for this enzyme in different species. The sequence of a number of mutations confirms the importance for enzyme activity of the carboxyl terminus of the protein. The most amino-terminal mutation (an ochre codon) results also in strikingly diminished steady-state levels of message. This agrees with results from other systems, suggesting that untranslated mRNA is preferentially degraded in eukaryotic cells. Northern blots show that specific induction, mediated by the UAY protein, and nitrogen metabolite repression, mediated by the AREA GATA factor, affect the steady-state levels of uaZ mRNA and thus act most probably at the level of transcription. PMID- 8226864 TI - A different combination of transcription factors modulates the expression of the human transferrin promoter in liver and Sertoli cells. AB - We have previously identified the functional regions involved in the regulation of human transferrin (Tf) gene expression in the liver and in Sertoli cells of the testis. Here, we show that a different cellular distribution of transcription factors, interacting with the same proximal promoter regions (PRI and PRII), modulates cell type-specific transcription. In the liver, hepatocyte nuclear factor 4 (HNF-4) and the chicken ovalbumin upstream promoter transcription factor (COUP-TF) act at the PRI site, while CCAAT/enhancer-binding proteins (C/EBPs) act at the PRII site. In the testis, distinct combinations of Sertoli proteins SP-A and SP-D and COUP-TF bind to the PRI site, while SP-alpha and SP-beta bind to the PRII site. Cotransfection experiments in Hep3B cells revealed that mostly HNF-4, C/EBP-alpha, C/EBP-delta, and, to a lesser extent, COUP-TF stimulated transcription driven by the -125/+39 region. In Sertoli cells, HNF-4 and COUP-TF appeared to repress, while the C/EBP factors were able to stimulate transcription driven by the -100/+39 region. However, the specific activating combination remains to be defined among the Sertoli proteins. In the non-Tf-expressing HeLa cells, the Tf promoter could be activated by C/EBP-delta. Our data revealed functional antagonism between HNF-4 and COUP-TF, binding to PRI, as well as cross coupling interactions between HNF-4 and C/EBP, binding to adjacent sites. Thus, cell type-specific DNA-protein interactions, together with protein-protein interactions, may explain the transcriptional regulation of the Tf gene in different cell types. PMID- 8226865 TI - Recovery of transcriptionally active chromatin restriction fragments by binding to organomercurial-agarose magnetic beads. A rapid and sensitive method for monitoring changes in higher order chromatin structure during gene activation and repression. AB - The unfolding of nucleosomes along transcriptionally active DNA sequences uncovers previously shielded cysteinyl-thiol groups of histone H3 molecules located at the center of the nucleosome core. This change in conformation and SH reactivity of nucleosomes along transcribed DNA sequences makes it possible to separate active from inactive nucleosomes by mercury affinity chromatography. The binding of thiol-reactive nucleosomes to an organomercurial-agarose column has been shown previously to reflect, with accuracy, both the timing and extent of transcription of the associated DNA sequences (Chen, T. A., and Allfrey, V. G. (1987) Proc. Natl. Acad. Sci. U. S. A. 84, 5252-5256). Here, we extend this experimental approach to the analysis of higher order chromatin structures. Large chromatin fragments released by treating isolated nuclei with restriction endonucleases are fractionated on mercurated agarose magnetic beads that capture nucleosomes with accessible histone H3 thiols, but do not react with the hidden H3 thiols of the compactly beaded nucleosomes of inactive genes. The SH-reactive domains of c-myc and other genes are rapidly separated from the non-SH-reactive restriction fragments by the magnetic bead technique. The new method also overcomes a major limitation of mercurated agarose column chromatography, which is not suitable for studies of higher order chromatin structure because large chromatin fragments occlude the mercury column; occlusion is not a problem in magnetic separations using suspended mercurated agarose beads. Here, we describe the synthesis of mercurated agarose magnetic beads with high capacity for SH groups and test their application to the recovery of chromatin restriction fragments of c-myc and the growth arrest gene gas1. PMID- 8226866 TI - Identification of the receptor subtype responsible for endothelin-mediated protein kinase C activation and atrial natriuretic factor secretion from atrial myocytes. AB - Endothelin-1 (ET-1) is a potent stimulator of atrial natriuretic factor (ANF) secretion from myocardial cells. In heart tissue there are two ET receptor subtypes (ETA-R and ETB-R), which can be pharmacologically distinguished by the ET isopeptides ET-1 and ET-3. However, the identification of the ET-R subtype responsible for the rapid enhancement of ANF release, which occurs within minutes of exposing cardiac myocytes to ET, has not been investigated. In the present study ET-1 was about 100-fold more potent than ET-3 at stimulating membrane phosphoinositide hydrolysis, protein kinase C activation, and ANF release from purified primary atrial myocytes. These responses were completely abolished by BQ123, an ETA-R antagonist. Radioligand binding analyses showed that competitor peptides displaced 125I-ET-1 binding to atrial myocyte ET-Rs with a rank order of potency of ET-1 >> BQ123 > ET-3, a characteristic ETA-R pharmacological profile. While neither ET-1 or ET-3 altered forskolin-stimulated cAMP levels, suggesting the absence of the ETB-R, basal cAMP levels were also unaffected by the ETs. Northern analysis using ET-R subtype-specific probes demonstrated that the ETA-R transcript was present in the cultures at levels at least 50-fold greater than the ETB-R transcript. These findings demonstrate that the stimulation of the phosphatidylinositol/protein kinase C pathway, which is required for maximal ET stimulated ANF release from primary atrial myocytes, is associated with the activation of only the ETA-R, thus defining a specific function for an endogenous ET-R in myocardial cells. PMID- 8226867 TI - Cloning of a novel human serotonin receptor (5-HT7) positively linked to adenylate cyclase. AB - An intron-containing gene encoding a novel human serotonin (5-HT) receptor was isolated from human genomic and cDNA libraries with probes directed to transmembrane regions of the adenylate cyclase stimulatory Drosophila serotonin receptor gene, 5-HTdrol. Membranes harvested from transiently transfected Cos-7 cells displayed high affinity (Kd = 8.5 nM), saturable (Bmax = 6.6 pmol/mg protein) [3H]5-HT binding. The rank order of potencies for serotonergic ligands to displace specific [3H]5-HT binding was: 5-carboxamido-tryptamine > methiothepin > metergoline > 5-HT > 8-hydroxy-2-(di-n-propylamino)tetralin > sumatriptan > ketanserin > zacopride. 5-HT produced a dose-dependent (EC50 = 992 nM) stimulation (approximately 20-fold) of cAMP accumulation in transiently transfected cells, and this response was antagonized by the nonselective 5-HT antagonist methiothepin. RNA for this gene was predominantly detected in the human brain and a subset of peripheral tissues including coronary artery and several tissues of the gastrointestinal tract. The molecular biological and pharmacological properties of this receptor suggest that it is the first member of a new serotonin receptor subfamily (5-HT7). The second messenger coupling, and tissue distribution indicate a possible identity to 5-HT receptors that mediate relaxant responses in certain isolated blood vessels. PMID- 8226868 TI - The replication fate of R- and S-styrene oxide adducts on adenine N6 is dependent on both the chirality of the lesion and the local sequence context. AB - In order to deduce the biological fate of adducts formed by the reaction of styrene oxide, a suspected carcinogen, with DNA, four oligodeoxynucleotides were synthesized which contained either R- or S-styrene oxide lesions on the N6 position of neighboring adenines within the human N-ras codon 61. When these adducted oligodeoxynucleotides were ligated into the single-stranded vector M13mp7L2 and the modified DNA used to transform repair-deficient Escherichia coli, the resultant plaque-forming abilities were found to vary as much as 300 fold, depending on the stereochemical configuration of the styrene oxide lesion and the sequence context in the vicinity of the damage. The frequency of mutations caused by the various styrene oxide adducts were similarly dependent on both their chirality and local sequence context. Oligodeoxynucleotide templates bearing these same four adducts were also constructed in order to evaluate their replication in vitro by the Klenow fragment. Three of the four styryl-modified templates yielded significant levels of fully extended primer upon polymerization. In contrast, the template containing R-styrene oxide at the second position of N-ras 61 was a very poor substrate for replication, a result which correlates well with the observed lethality of this lesion in vivo. PMID- 8226869 TI - Ethanol differentially increases alpha 2-adrenergic and muscarinic acetylcholine receptor gene expression in NG108-15 cells. AB - Modulation of alpha 2-adrenergic and opioid neurotransmission may contribute to ethanol intoxication, tolerance, and physical dependence. We showed previously that ethanol increased the expression of functional delta-opioid receptors in NG108-15 cells (Charness, M. E., Querimit, L. A., and Diamond, I. (1986) J. Biol. Chem. 261, 3164-3169). Here we report that long-term (2 days) treatment of NG108 15 cells with ethanol increased the binding of the alpha 2-adrenergic receptor (alpha 2AR) antagonist [3H]rauwolscine and the muscarinic acetylcholine receptor (mAChR) antagonist [3H]quinuclidinyl benzilate by 2.8- and 1.4-fold, respectively. Increased receptor expression was associated with a proportionate increase in the potency of oxymetazoline and carbachol in inhibiting cAMP accumulation. Ethanol did not change the expression of G alpha i2 and reduced levels of G alpha s. Pertussis toxin pretreatment did not prevent the ethanol induced increase in alpha 2AR, mAChR, and delta-opioid receptor expression. Ethanol caused a large (3.6-fold), dose-dependent increase in the abundance of alpha 2BAR mRNA (rat cDNA probe RNG, 4.1-kb transcript). Ethanol-induced increases in alpha 2BAR and alpha 2CAR (rat probe RG10, 2.5-kb transcript) mRNAs were first detected after 6 h of exposure to 100 mM ethanol, became maximal after 24 h, and persisted for up to 5 days. In contrast, ethanol caused only a small (1.3-fold) increase in the abundance of hm4 mAChR mRNA and did not change levels of G alpha i2 and G alpha s mRNAs. Our data indicate that clinically attainable concentrations of ethanol regulate alpha 2AR gene expression within the time frame of a single session of drinking. PMID- 8226870 TI - Demonstration of an inducible cyclooxygenase in human endothelial cells using antibodies raised against the carboxyl-terminal region of the cyclooxygenase-2. AB - Cyclooxygenase (Cox) exists in two forms in human endothelial cells (HUVEC). We have raised antibodies that recognize the sequence of the carboxyl-terminal portion of the human Cox-2 (C)-NASSSRSGLD-DINPTVLLK. Cyclooxygenase activity of HUVEC challenged with interleukin 1 alpha or a phorbol ester increased in parallel with the mass of a protein doublet analyzed by Western blot using antibodies directed against the Cox-2 peptide; a monoclonal antibody directed against Cox-1 showed a small change in protein mass. A 35S-labeled protein doublet with a molecular mass of approximately 70,000 daltons was immunoprecipitated with the anti-Cox-2 antiserum in L-[35S] methionine-labeled cells stimulated with interleukin 1 alpha. This protein was not recovered by pretreating the antiserum with the Cox-2 peptide before immunoprecipitation. A minor variation in 35S-immunoprecipitated protein was obtained with the polyclonal anti-Cox-1 antibody. Both immunoprecipitated Cox-1 and Cox-2 possessed cyclooxygenase activity that was inhibited by flurbiprofen. Endoglycosidase H treatment of immunoprecipitated Cox-2 proteins caused a decline in the apparent molecular size similar to that observed with immunoprecipitated Cox-1 or sheep cyclooxygenase but did not suppress the doublet. These results show by direct protein measurement that HUVEC synthesize the novel Cox-2 under appropriate stimulation, with little changes of Cox-1. PMID- 8226871 TI - Intramolecular fusion of Na pump subunits assures exclusive assembly of the fused alpha and beta subunit domains into a functional enzyme in cells also expressing endogenous Na pump subunits. AB - Experiments designed to identify Na pump structural features which tag the molecule for asymmetric cell-surface localization are inherently complex because either subunit, or both, may contain targeting information and because the cells which recognize those targeting signals and maintain asymmetric plasma membrane domains also express their own Na pumps, the subunits of which can assemble into hybrid pump molecules with pump subunits expressed from transfected cDNA clones. Cotransfecting cDNA for both subunits only complicates matters further by resulting in expression of four distinct dimeric molecular species. To eliminate the potential for cross-assembly in these and other experiments we have constructed cDNA encoding a "single-subunit" Na pump (called "alpha-beta") in which the alpha and beta subunits are joined by a linker of 17 amino acids. By all criteria tested alpha-beta functioned as a normal heterodimeric Na pump. It was expressed in a variety of mammalian cell lines as a single, high molecular weight polypeptide located primarily on the surface membrane, with the beta subunit exposed to the extracellular medium. Binding of the conformation sensitive monoclonal antibody 24 to the beta subunit indicated that the fusion protein was folded as a properly "assembled" sodium pump. Expression of alpha beta in ouabain-resistant mouse L cells resulted in high affinity ouabain binding and ouabain-sensitive, sodium-dependent rubidium transport. The enzyme was properly targeted to the basolateral plasma membrane in polarized epithelial cells. The functional integrity of the fusion protein renders it suitable for site-directed mutagenesis studies of targeting and enzymology where control of subunit assembly is desired. These results also support topological models in which the carboxyl terminus of the alpha subunit is cytoplasmic. PMID- 8226872 TI - v-Src activates the expression of 92-kDa type IV collagenase gene through the AP 1 site and the GT box homologous to retinoblastoma control elements. A mechanism regulating gene expression independent of that by inflammatory cytokines. AB - The 92-kDa type IV collagenase (matrix metalloproteinase-9; MMP-9) is frequently expressed in cells showing an invasive nature during physiological and pathological processes, and the expression is strictly controlled by a variety of trans-membrane signals. Binding sites for NF-kB, Sp-1, and AP-1 are reportedly required for induction of MMP-9 gene expression by tumor necrosis factor-alpha or 12-O-tetradecanoylphorbol-13-acetate. Comparison of the sequence of the newly cloned mouse MMP-9 promoter region with our previous human isolate revealed that, in addition to the above mentioned elements, four units of GGGG(T/A)GGGG sequence (GT box) were conserved between the two species. In this study, we have demonstrated that one of the GT boxes located downstream of the AP-1 site is essential along with the AP-1 site for the activation of the promoter by v-Src but not by tumor necrosis factor-alpha or 12-O-tetradecanoylphorbol-13-acetate. Gel mobility-shift assays revealed that binding proteins for retinoblastoma control element, including Sp-1 family protein, can bind specifically to GT boxes. Thus, the v-Src signals to the AP-1 site and to the GT box homologous to retinoblastoma control element acted synergistically in transcriptional activation. These results suggest that certain v-Src-mediated signals are propagated along pathways that are independent of inflammatory cytokines. PMID- 8226873 TI - Aldosterone-mediated Na/K-ATPase expression is alpha 1 isoform specific in the renal cortical collecting duct. AB - In the renal cortical collecting duct (CCD), mineralocorticoid hormones, like aldosterone, augment the abundance of Na/K-ATPase molecules. It has been postulated that this response involves an isoform switch of the Na/K-ATPase catalytic subunit, alpha, as the molecular basis for the differential regulation of mineralo-corticoid-induced and constitutively expressed Na/K-ATPase pools. In opposition to this attractive hypothesis, three lines of independent evidence are presented which demonstrate that the CCD exclusively expresses the alpha 1 form despite mineralocorticoid-mediated changes in functional Na/K pump density. First, aldosterone increased [3H]ouabain binding in CCD 2.5-fold without changing the ouabain dissociation constant. Second, an electrophysiological assay for pump activity revealed that aldosterone increased maximum Na/K pump current in parallel with the change in ouabain binding without altering the apparent sodium affinity. Third, Western blot analysis with alpha isoform-specific, antipeptide antibodies demonstrated that aldosterone exclusively increased the total chemical pool of the alpha 1 form of the pump without inducing other alpha subunit isoforms. In summary, aldosterone increases the abundance of Na/K-ATPase molecules in the CCD which are pharmacologically, physiologically, and chemically indistinguishable from those that are normally expressed. PMID- 8226874 TI - Two modes of transcription initiation in vitro at the rrnB P1 promoter of Escherichia coli. AB - The rrnB P1 promoter of Escherichia coli (starting sequence C-4-A-3-C-2-C-1-A+1 C+2-U+3-G+4) forms a binary complex with RNA polymerase that is highly unstable and requires the presence of transcription substrates ATP and CTP for stabilizing the enzyme-DNA association (Gourse, R. L. (1988) Nucleic Acids Res. 16, 9789 9809). We show that in the absence of UTP and GTP the stabilization is accomplished by short RNA oligomers synthesized in an unusual "-3-->" mode whereby the primer initiated at the +1 site presumably slips back by three nucleotides into the -3 site and is then extended yielding stable ternary complexes. By contrast, short oligomers initiated in the conventional "+1-->" mode without slippage do not exert the stabilization effect and are readily aborted from the promoter complex. The stable -3-->ternary complexes carry sigma factor but otherwise resemble elongation complexes in their high salt stability and in the fact that they are formed with a mutant RNA polymerase deficient in promoter binding. A model is proposed explaining the stability of the -3- >ternary complexes by RNA slipping into a putative "tight RNA binding site" in RNA polymerase which is normally occupied by RNA during elongation. PMID- 8226875 TI - Reciprocal regulation of mRNA and protein for subunits of cAMP-dependent protein kinase (RI alpha and C alpha) by cAMP in a neoplastic B cell line (Reh). AB - The present study examines the activity, levels of expression and regulation of cAMP-dependent protein kinase subunits during cAMP-mediated inhibition of Reh cell proliferation. Human Reh cells express mRNAs for the RI alpha and C alpha subunits of cAK at high levels and are practically devoid of cAMP-dependent protein kinase type II. Treatment with isoproterenol, forskolin, or a cAMP analog increased RI alpha mRNA in a time- and concentration-dependent manner (maximal, 4 fold, at 4-8 h). Messenger RNA for C alpha was also stimulated by cAMP, although with slower kinetics (maximal, 2-fold, at 16-24 h). Nuclear run-on assays showed a 2-fold increase in RI alpha gene transcription, whereas that of C alpha was unchanged. In spite of the stimulatory effects of cAMP on mRNAs for both RI alpha and C alpha, phosphotransferase activity and specific [3H]cAMP binding decreased rapidly after treatment with either cAMP or forskolin. Interestingly, the decrease in R and C activity preceded the increase in RI alpha and C alpha mRNA levels, raising the question whether increased mRNA levels may be secondary to the decrease in RI alpha or C alpha protein. The finding that the protein synthesis inhibitor cycloheximide gave changes in RI alpha and C alpha mRNA similar to cAMP and that co-treatment with cycloheximide and cAMP resulted in additive effects tend to support this notion. PMID- 8226876 TI - Polypeptide import and degradation by isolated lysosomes. AB - We reported previously that lysosomes derived from human diploid fibroblasts import and degrade polypeptides and that these processes are stimulated by ATP and by the heat shock cognate protein of 73 kDa (hsc73). We now report several new aspects of this in vitro proteolytic pathway. (a) Among four polypeptides tested, this pathway appears to be selective for those containing KFERQ-like peptide motifs. (b) Substrate proteins specifically bind to a protein-containing site on lysosomal membranes. (c) Lysosomes derived from serum-deprived cells are twice as active as those from serum-supplemented cells. (d) A portion of intracellular hsc73 is associated with lysosomes, and the amount of lysosomal hsc73 increases in response to serum withdrawal. Additional characterization of this proteolytic pathway not reported previously shows that intact lysosomes are required, the import process is saturable with an apparent Km of 5 microM for RNase S-peptide, and reducing agents activate this lysosomal import and degradation pathway. PMID- 8226877 TI - Isolation of two distinct 3-hydroxysteroid sulfotransferases from the guinea pig adrenal. Evidence for 3 alpha-hydroxy versus 3 beta-hydroxy stereospecificity. AB - Two physically distinct hydroxysteroid sulfotransferases (HSTs) that demonstrate substrate specificity with respect to the orientation of the ring A 3-hydroxyl group have been isolated from the guinea pig adrenal gland. Nondenaturing liquid phase isoelectric focusing permitted the separation of the activities, and reverse phase high performance liquid chromatography was used to purify the two proteins to homogeneity. The 3 beta-HST had an apparent molecular mass of 33 kDa and utilized pregnenolone, 17-hydroxypregnenolone, and dehydroepiandrosterone as substrates. The 3 alpha-HST was slightly smaller at 32 kDa and utilized allopregnanolone and androsterone as substrates. The proteins were further distinguished by isoelectric point, immunoreactivity, and tryptic peptide mapping. Peptides isolated from both guinea pig HSTs demonstrated significant amino acid sequence homology (approximately 65% identity) to rat liver HST; however, available sequence data from the two proteins did not yield differences that might account for their stereospecific substrate selectivity. This paper represents the first definitive report demonstrating the existence of discrete HSTs that exhibit substrate specificity based on the stereochemistry of the 3 hydroxyl group. PMID- 8226878 TI - cDNA cloning of chick cartilage chondroitin sulfate (aggrecan) core protein and identification of a stop codon in the aggrecan gene associated with the chondrodystrophy, nanomelia. AB - We previously reported the cloning and sequencing of a 1.5-kilobase cDNA which encoded a portion of the chondroitin sulfate domain from the chick cartilage proteoglycan core protein and the localization of a species-specific monoclonal antibody epitope. Using polymerase chain reaction amplification and primer extension, cDNA clones which code for the entire proteoglycan core protein have now been obtained from a 10-day chick embryo cDNA library. The composite sequence is 6464 nucleotides long, coding for a protein of 2109 amino acid residues with a calculated M(r) = 223,500. The overall arrangement of globular and carbohydrate attachment domains is similar to human and rat chondrosarcoma aggrecan, but there are significant differences in detailed homology between chick and mammalian core proteins. Most significantly a highly repetitive region (19 repeat units of 20 residues each), not found in either human or rat, enlarges one of the characteristic serine-glycine containing regions (designated CS-2) while the other serine-glycine containing domain (designated CS-1) is approximately one fourth the length of the mammalian CS-1. Analysis of a polymerase chain reaction amplified fragment encoding the chick-specific repeat region revealed a single base mutation at position 4553 (G to T transversion) that converted the codon GAA for glutamate at amino acid 1513 to TAA, a stop codon, in nanomelic chondrocytes. Genomic DNA from nanomelic liver was also digested with restriction enzyme BsaBI to verify the G to T transversion. This single mutation leads to a shortened core protein precursor with a calculated M(r) = 158,300. The resulting phenotype, nanomelia, arises because the truncated core protein is neither processed to a mature proteoglycan, nor secreted from the chondrocyte. PMID- 8226879 TI - Brain proline-directed protein kinase phosphorylates tau on sites that are abnormally phosphorylated in tau associated with Alzheimer's paired helical filaments. AB - Brain proline-directed protein kinase (BPDK), which contains a catalytic subunit homologous to and displaying site-specific phosphorylation similar to p34cdc2 kinase (Lew, J., Winkfein, R. J., Paudel, H. K., and Wang, J. H. (1992) J. Biol. Chem. 267, 25922-25926), has been examined for possible involvement in tau phosphorylation. Immunoblot analyses using peptide antibodies specific for BPDK have revealed the presence of the kinase in bovine brain microtubules purified extensively by repeated polymerization and depolymerization cycles. When the microtubule proteins are separated into the tubulin and microtubule-associated protein fractions, BPDK is found exclusively in the latter fraction. BPDK phosphorylates both tau and MAP2, the former protein being phosphorylated to a stoichiometry of 3.8 mol of phosphate/mol of tau. Analysis of the phosphopeptides isolated from the tryptic digest of the phosphorylated bovine tau has revealed seven phosphorylation sites. Based on the sequence alignment between bovine and human tau proteins, these sites correspond to Ser-195, Ser-202, Thr-205, Thr-231, Ser-235, Ser-396, and Ser-404 of human tau. Mass spectrometric analysis of the tau protein isolated from Alzheimer's paired helical filaments (PHFs) has determined three abnormal phosphorylation sites and two phosphopeptides containing a total of five abnormal phosphates (Hasegawa, M., Morishima Kawashima, M., Takio, K., Suzuki, M., Titani, K., and Ihara, Y. (1992) J. Biol. Chem. 267, 17047-17054). Two of the sites in tau phosphorylated by BPDK, Thr-231 and Ser-235, are among the abnormal phosphorylation sites, and the other sites phosphorylated by BPDK are within phosphopeptides from PHF-tau. These results suggest that BPDK may be one of the kinases responsible for the abnormal phosphorylation-associated PHF-tau. PMID- 8226880 TI - Reconstitution of recombinant 40-kDa subunit of the clathrin-coated vesicle H(+) ATPase. AB - We have proposed a model of the ATP hydrolytic sector of the clathrin-coated vesicle H(+)-ATPase wherein significant catalysis requires four subunits of molecular masses of 70, 58, 40, and 33 kDa (Xie, X.-S., and Stone, D. K. (1988) J. Biol. Chem. 263, 9859-9867). We have cloned and expressed the 40-kDa component in Escherichia coli and have purified the recombinant protein to homogeneity. This subunit lacks ATP hydrolytic capacity, but when reconstituted to a 40 kDa depleted hydrolytic sector, there is a greater than 20-fold increase in calcium activated, N-ethylmaleimide-sensitive ATP hydrolysis, indicating that this subunit is required for vacuolar-type proton pump function. PMID- 8226881 TI - Cloning and expression of cDNA for a human enzyme that hydrolyzes 8-oxo-dGTP, a mutagenic substrate for DNA synthesis. AB - 8-Oxoguanine (8-oxo-7, 8-dihydroguanine) is produced in DNA, as well as in nucleotide pools of cells, by active oxygen species normally formed during cellular metabolic processes. 8-Oxoguanine nucleotide can pair with cytosine and adenine nucleotides at almost equal efficiencies, and transversion mutation ensues. Human cells contain enzyme activity, which hydrolyzes 8-oxo-dGTP to 8-oxo dGMP, and this enzyme is responsible for preventing misincorporation of 8 oxoguanine into DNA. We purified this particular human enzyme to physical homogeneity and determined a partial amino acid sequence. We then cloned the cDNA for human 8-oxo-dGTPase and examined its nucleotide sequence. The human protein comprises 156 amino acid residues and has some sequence homology with the Escherichia coli MutT protein, which has a distinct 8-oxo-dGTPase activity. When the human cDNA was expressed in E. coli mutT- mutant cells, there was a significant amount of 8-oxo-dGTPase activity. In such cells, the frequency of spontaneous mutation was greatly reduced. We propose that the human 8-oxo-dGTPase protects genetic information from the untoward effects of endogenous oxygen radicals. PMID- 8226882 TI - Repression of c-fos gene expression by thyroid hormone and retinoic acid receptors. AB - Incubation with 1 nM triiodothyronine (T3) decreased cycloheximide-induced c-fos mRNA levels and the mRNA response to the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate (TPA; 100 nM) or to forskolin (15 microM). T3 also reduced the abundance of nuclear proteins that bind to an AP-1 binding site and the levels of c-Fos protein, as determined by Western blot. In transient transfection assays with a c-fos promoter reporter construct T3 decreased basal promoter activity by more than 50-60% and strongly inhibited the TPA and forskolin-induced activity. T3 further suppressed basal promoter activity and led to a drastic decrease of the response to both stimulatory agents after co-transfection of the promoter with an expression vector for the receptor. A truncated receptor that lacks the DNA binding domain abolished the inhibitory effect of the endogenous and transfected T3 receptor. Retinoic acid (RA) had an effect similar to T3 reducing basal CAT activity and the response to TPA and forskolin by 40-50% in cells co transfected with the c-fos promoter and an expression vector for the RA receptor. The inhibitory effect of the nuclear receptors on c-Fos is reciprocal, since overexpression of c-Fos repressed induction of the activity of the growth hormone promoter by T3 and RA. Co-transfection with an antisense c-fos vector relieved this inhibition and restored the response to both ligands. These results show the antagonism between the nuclear receptors and the membrane signal transduction pathways that converge on the c-fos oncogene. PMID- 8226883 TI - Activator region analysis of the human D1A dopamine receptor gene. AB - The human D1A dopamine receptor gene is transcribed from a TATA-less promoter, and its main activator is between nucleotides -1342 and -1102 relative to the first ATG (Minowa, M. T., Minowa, T., Monsma, F. J., Jr., Sibley, D. R., and Mouradian, M. M. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 3045-3049). In the present study, additional serial deletion mutants fused with a reporter gene and used in transient transfections of the D1A-expressing cell line NS20Y further localized this activator to two regions: -1154 to -1137, which increases promoter activity 4-fold, and -1197 to -1154, which confers an additional 2-fold increase in transcriptional activity. Cell specificity of the D1A gene expression is contributed by both the -1197/-1154 region and the core promoter located downstream to -1102. DNase I footprinting and gel shift assays suggested that DNA protein interactions occur primarily between -1197 and -1116. Competitive cotransfections using different portions of this segment, D1A-Act-1 (-1197 to 1152) and D1A-Act-2 (-1154 to -1116), confirmed the in vivo functional significance of these sequences in activating the D1A promoter. D1A-Act-1 has no consensus sequences for known transcription factors but binds to several proteins as a complex. Although D1A-Act-2 has AP2 consensus sequences and binds to recombinant AP2, the nuclear factors in NS20Y cells interacting with this region are distinct from AP2 itself. Despite the inability of Sp1 to directly bind to its consensus sequences in D1A-Act-2, Sp1 or another protein antigenically related to Sp1, as well as a novel nuclear factor, is included in the complex that binds to this activator. PMID- 8226884 TI - A phenylalanine codon deletion at the UGT1 gene complex locus of a Crigler-Najjar type I patient generates a pH-sensitive bilirubin UDP-glucuronosyltransferase. AB - The characterization (Ritter, J. K., Chen, F., Sheen, Y. Y., Tran, H. M., Kimura, S., Yeatman, M. T., and Owens, I. S. (1992) J. Biol. Chem. 267, 3257-3261) of the single-copy UGT1 gene complex encoding both bilirubin and phenol UDP glucuronosyltransferases (transferase) has been critical to the determination of genetic defects in Crigler-Najjar Type I patients. The complex (UGT1A-UGT1G) codes for at least two bilirubin, three bilirubin-like, and two phenol transferases. Seven different exons 1, each with an upstream promoter and each encoding the amino terminus of an isoform, are arrayed in series with four common exons (encoding seven identical carboxyl termini) in the 3'-region of the locus. Predictably, a critical mutation in a common exon inactivates the entire locus. A deleterious mutation in an exon 1, as we report here for the UGT1A gene in a Crigler-Najjar Type I patient, predictably affects the amino terminus of that single isoform. The code for the predominant bilirubin isozyme, the HUG-Br1 protein, is missing the phenylalanine codon at position 170 in exon 1 of UGT1A, abolishing a conserved diphenylalanine. We demonstrate that, at the pH (7.6) routinely used for bilirubin glucuronidation studies, both the HUG-Br1 protein and human liver microsomes have approximately one-third the activity seen at the major pH optimum of 6.4 and at low ionic strength. The altered isozyme with nearly normal activity at pH 7.6 is inactive at pH 6.4, a result consistent with the definition of a pH-sensitive mutant. The Km value for bilirubin using the wild-type protein is approximately 2.5 microM at both pH 6.4 and 7.6 and that for the mutant is 5.0 microns at pH 7.6. The structure of the wild-type enzyme compared to that of the mutant indicates that hydrophobic properties at the active center are critical for metabolizing the lipophile-like substrate. The low ion/pH requirements for bilirubin glucuronidation may signal the basis for the distribution of these isozymes to an organelle (endoplasmic reticulum) that can establish compatible conditions/compartments for each catalysis. PMID- 8226885 TI - Calcium-dependent activation of protein kinase C. The role of the C2 domain in divalent cation selectivity. AB - Activation of certain isoforms of protein kinase C (cPKCs) requires Ca2+ and is associated with a conserved C2 domain that is not present in Ca(2+)-independent isoforms (nPKCs). The site(s) of Ca2+ binding and the role of the C2 domain have not been previously identified. We have analyzed phosphatidylserine-dependent Ca2+ binding to fusion proteins expressed in Escherichia coli that carry various modifications in the regulatory region of cPKC beta 1 or nPKC epsilon. Ca2+ is bound mainly to the C1 domain of PKC beta 1, but the C2 domain confers specificity for Ca2+ binding when compared with Mg2+ and Mn2+. We propose that in cPKCs there is selective binding of Ca2+ to a pocket formed by the C1 and C2 domains. This induces a change in conformation that activates the enzyme. In nPKCs, the cation binding pocket is less specific for Ca2+ because it lacks the C2 domain. Therefore, divalent cations like Mg2+ can bind to it, thereby abrogating the requirement of Ca2+ for enzyme activation. PMID- 8226886 TI - Calpain activity increases in hepatocytes following addition of ATP. Demonstration by a novel fluorescent approach. AB - Our aim was to measure calpain protease activity during increases in cytosolic free calcium (Ca2+i) after addition of extracellular ATP. The calpain protease substrate t-butoxycarbonyl-Leu-Met-7-amino-4-chloromethylcoumarin was synthesized. Nonfluorescent t-butoxycarbonyl-Leu-Met-7-amino-4- chloromethyl coumarin diffuses into the cell where it is conjugated to glutathione forming t butoxycarbonyl-Leu-Met-7-amino-4-methylcoumarin glutathione conjugate (Boc-Leu Met-MAC-SG). The nonfluorescent, membrane impermeant Boc-Leu-Met-MAC-SG accumulates in the cell. Intracellular proteolytic hydrolysis of Boc-Leu-Met-MAC SG releases and unquenches the fluorescence of MAC-SG. Intracellular fluorescence of MAC-SG was quantitated in single, cultured rat hepatocytes using digitized video fluorescent microscopy. Enhancement of intracellular fluorescence generation by increases in Ca2+i and inhibition by a calpain inhibitor indicated the probe was a calpain substrate. After addition of ATP, calpain protease activity increased to 156 +/- 13% of basal concurrent with a 3-fold rise of Ca2+i for 2-4 min. Thereafter, Ca2+i decreased to values of 1.5-fold above basal and protease activity returned to normal. Incubation of cells in Ca(2+)-free buffer abolished the rise in Ca2+i and calpain protease activity. Calpain protease activity increases concomitantly with increases of Ca2+i supporting the hypothesis that calpain proteases participate in Ca(2+)-mediated signal transduction. PMID- 8226887 TI - Imaging of inositol 1,4,5-trisphosphate-induced Ca2+ fluxes in single permeabilized hepatocytes. Demonstration of both quantal and nonquantal patterns of Ca2+ release. AB - The subcellular organization and function of inositol 1,4,5-trisphosphate (InsP3) sensitive Ca2+ stores has been investigated in permeabilized hepatocytes using fluorescent probe techniques that monitor InsP3 action at the level of the Ca2+ storage organelles. Chlortetracycline fluorescence was used to follow alterations in luminal Ca2+, and InsP3-activated Mn2+ quench of compartmentalized fura-2 was used as a measure of the distribution and permeability of the InsP3-sensitive channels. Fluorescence imaging of single permeabilized hepatocytes attached to coverslips demonstrated that InsP3-sensitive Ca2+ stores are distributed throughout the cytoplasm, but are not present within the nuclear matrix. When hepatocytes were permeabilized in suspension, InsP3 activation of channel opening and Ca2+ release occurred in a quantal manner, such that the incremental magnitude of the response was determined by the dose of InsP3. Under these conditions dose-dependent steps of InsP3-induced Mn2+ entry into the stores occurred in the absence of changes in cytosolic or luminal Ca2+, providing evidence for a series of separate compartments with different sensitivities to InsP3. Electron microscopy studies revealed that the endoplasmic reticulum was extensively vesicularized when hepatocytes were permeabilized in suspension, whereas essentially normal endoplasmic reticulum structure was retained in cells attached to coverslips. In these attached cells the Ca2+ release and channel opening responses to InsP3 occurred in a nonquantal manner at the single cell level. Submaximal doses of InsP3 gave the same magnitude of response as a maximal InsP3 dose, although the rates of Ca2+ release and Mn2+ permeation through the InsP3-activated channels increased in a dose-dependent manner. Thus, in each cell the entire Ca2+ store was accessible for mobilization by all effective InsP3 concentrations. We conclude that the quantal release properties of the InsP3 receptor are not expressed in attached permeabilized liver cells because there is extensive luminal continuity within the InsP3-sensitive Ca2+ stores. This continuity appears to be disrupted when hepatocytes are permeabilized in suspension. PMID- 8226888 TI - A site on transducin alpha-subunit of interaction with the polycationic region of cGMP phosphodiesterase inhibitory subunit. AB - Activation of cGMP phosphodiesterase (PDE) by the rod G-protein transducin is a key event in visual signal transduction in vertebrate photoreceptor cells. Interaction between the GTP-bound form of the alpha-subunit of transducin (alpha t*) and the PDE inhibitory gamma-subunit (P gamma) is a major component of PDE activation. The central polycationic region of P gamma, P gamma-24-45, has been implicated as one of the sites involved in alpha t*.P gamma interaction. Here we determine the site on alpha t* that interacts with P gamma-24-45 using a photo cross-linking approach. The synthetic peptides Cys(ACM)Tyr-P gamma-24-45-Cys (where ACM indicates acetamidomethyl group) and Cys-P gamma-24-45 were labeled with 4-(N-maleimido)benzophenone at the COOH and NH2 termini, respectively, and then cross-linked to alpha t. When the photoprobe was attached to the COOH terminus of the peptide, a specific high yield cross-linked product (80%) was formed between the peptide and alpha t GTP gamma S (guanosine 5'-O (thiotriphosphate)). A lower yield of cross-linking (35%) was seen between the peptide and alpha t GDP. The site of cross-linking between Cys(ACM)Tyr-P gamma-24 45-Cys and alpha t GTP gamma S was localized to within alpha t-306-310 using a variety of chemical and proteolytic cleavages of the cross-linked product, analysis of the fragments with SDS-polyacrylamide gel electrophoresis, and matrix assisted laser desorption ionization mass spectrometry. PMID- 8226889 TI - Reaction of human chymase with reactive site variants of alpha 1 antichymotrypsin. Modulation of inhibitor versus substrate properties. AB - Inhibition of human chymase by alpha 1-antichymotrypsin produces 3.5 mol of degraded inhibitor for every mol of chymase inhibited, resulting in a stoichiometry of inhibition (SI) of 4.5. In the present study, the substrate versus inhibitor properties of this reaction were examined further using wild type and mutant recombinant antichymotrypsins (rACT). Titration of chymase hydrolytic activity with rACT-L358 (wild type) and reactive site (P1) variants of ACT, L358W, L358M, and L358F revealed that the SI was sensitive to P1 residue replacements. SI values increased in the order of Trp < Met < Leu < Phe where SI values were 1.5, 2, 4, and 7, respectively. Chymase inhibitor complex and cleaved inhibitor were demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for all variants; the relative intensities of each band were consistent with SI values established by titration. NH2-terminal sequence analyses of the products formed in the reaction of chymase with rACT-L358F indicated that the P1-P1' bond was the primary site of cleavage resulting in the hydrolysis and inactivation of this variant. The apparent second-order rate constant for chymase inhibition (k'/[I]) by rACT also was affected by P1 substitution. k'/[I] values increased in an order opposite that obtained for SI values (Phe < Leu < Met < Trp). The reactive loop mutant (rACT-P3P3') produced by replacing the reactive site region of ACT (Thr356-Val361) with that of alpha 1 proteinase inhibitor (Ile356-Pro361) revealed a different reaction pattern. Although its SI was near 1, the value for k'/[I] was the lowest among variants. rACT-L358R, another P1 variant, did not inhibit chymase. These results are evaluated with respect to the substrate preferences of human chymase and with respect to partitioning schemes proposed to explain SI values greater than 1. PMID- 8226890 TI - Overexpression of YAP2, coding for a new yAP protein, and YAP1 in Saccharomyces cerevisiae alleviates growth inhibition caused by 1,10-phenanthroline. AB - A new member of the yeast yAP family, designated YAP2, has been isolated and characterized. The protein displays a high homology with the DNA-binding domain of yAP-1, which contains a basic DNA-binding domain and leucine zipper motif and binds in vitro to the same cis-element. Growth arrest of yeast caused by low concentrations of 1,10-phenanthroline, resulting in zinc and/or iron deprivation, is overcome by over-expression of YAP1 or YAP2. In fact, yeast cells over expressing YAP1 or YAP2 display pleiotropic drug resistance. On the other hand, a yap2 null mutant has an increased thermotolerance under starvation conditions caused by 1,10-phenanthroline. The latter mutant might become an excellent tool in the study of pathways leading toward thermotolerance acquisition. PMID- 8226891 TI - Functional domain in an arginine-rich carboxyl-terminal region of p47phox. AB - Activation of the neutrophil respiratory burst oxidase involves phosphorylation dependent translocation of the cytosolic proteins p47phox and p67phox to the plasma membrane, a process in intact cells that is inhibited by staurosporine. We now report that in a cell-free oxidase system, staurosporine and protein kinase C pseudosubstrate PKC(19-36) both inhibited p47phox phosphorylation but had no effect on superoxide generation. In contrast, p47phox phosphorylation, translocation, and superoxide generation were inhibited by a peptide, p47phox(323 332) (AYRRNSVRFL), based on a putative serine phosphorylation domain. This effect was specific for the 323-332 peptide, as it was not observed with two peptides based on other p47phox phosphorylation domains. All three peptides served as substrates for phosphorylation, but the extent of peptide phosphorylation did not correlate with inhibition of oxidase function. p47phox(325-330), which represents the serine phosphorylation motif of p47phox(323-332), did not inhibit translocation or superoxide production despite its ability to block phosphorylation of p47phox. These data indicate the presence of functionally important sites within the p47phox(323-332) peptide. Although serine 328 is in a consensus phosphorylation motif, the lack of correlation in the cell-free system between p47phox phosphorylation and either protein translocation or superoxide formation suggests that a phosphorylation-independent function resides in the 323 332 segment of p47phox. PMID- 8226892 TI - Hormone response complex in a novel abscisic acid and cycloheximide-inducible barley gene. AB - The phytohormone, abscisic acid (ABA), plays a variety of roles during seed development and in the plant's response to environmental stresses. To study the molecular action of ABA, we have isolated a single copy ABA-induced gene, HVA22, which is mapped to barley chromosome 1. The HVA22 gene can be induced by either ABA or the protein synthesis inhibitor, cycloheximide, and addition of both inducers to barley aleurone layers has a synergistic effect on the expression of this gene. Sequence comparison indicates that the HVA22 gene product is highly homologous to the product of human DP1 gene, which is likely to contribute to colorectal tumorigenesis. The hormonal regulation of HVA22 expression has been studied, and there appear to be at least three elements, two located in the promoter and one in the first intron, which are essential for the high level of ABA induction of HVA22 expression. Among the promoter elements is a homolog of ABA response element, which has been shown to be important in the expression of other ABA-induced genes in plants. We suggest that the barley HVA22 gene product is likely a regulatory protein, and the ABA induction of this gene requires the action of a complex set of hormone response elements. PMID- 8226893 TI - Dinitrogenase reductase- and MgATP-dependent maturation of apodinitrogenase from Azotobacter vinelandii. AB - The requirements for iron-molybdenum cofactor (FeMo-co) activation of apodinitrogenase from Azotobacter vinelandii strain UW97, which lacks dinitrogenase reductase activity as assayed by substrate reduction, have been examined. Activation of apodinitrogenase from strain UW97 by FeMo-co requires the addition of both dinitrogenase reductase and MgATP. When the same apodinitrogenase is pretreated with dinitrogenase reductase and MgATP and then partially purified, however, it does not require these components for activation by FeMo-co. This suggests that dinitrogenase reductase and MgATP are involved in processing apodinitrogenase to a FeMo-co activatable form. This processing step coincides with a change in the subunit composition of apodinitrogenase from alpha 2 beta 2 to a form with an additional subunit (gamma) attached. The apodinitrogenase with the associated gamma subunit is apparently the form of the protein that is competent for activation by FeMo-co. PMID- 8226894 TI - Structural diversity in the alpha 2-->8-linked polysialic acid chains in salmonid fish egg glycoproteins. Occurrence of poly(Neu5Ac), poly(Neu5Gc), poly(Neu5Ac, Neu5Gc), poly(KDN), and their partially acetylated forms. AB - alpha 2-->8-Linked polysialic acid (polySia) chains terminate O-linked oligosaccharide chains on Salmonidae fish egg polysialoglycoproteins (PSGPs). Expression of these surface PSGPs are developmentally regulated and the polySia epitope is functionally implicated in a number of distinct species-specific cell cell recognition events during fertilization and early embryogenesis. To better understand the functional diversity of these PSGPs, structural studies of the polySia chains isolated from three genera and eight species of Salmonidae fish eggs were carried out by chemical, immunochemical, enzymatic, and 1H NMR methods. A remarkable degree of structural diversity was found, including differences in the N-acyl groups, i.e. N-acetylneuraminic acid (Neu5Ac) or N-glycolylneuraminic acid (Neu5Gc), and in the presence of either O-acetyl substitution at C4, C7, or C9 or O-lactyl substitution at C9. The presence of heteropolymers containing both Neu5Ac and Neu5Gc residues was also an unexpected finding. Accordingly, the different forms of alpha 2-->8-linked homo- and heteropolymers of these polySia structures include: poly(Neu5Ac), poly(Neu5Gc), poly(Neu5,chi Ac2), poly(Neu5Gc chi Ac), poly(Neu5Ac, Neu5Gc), poly-(Neu5Ac, Neu5,chi Ac2), poly(Neu5Ac, Neu5Gc chi Ac), poly(Neu5Gc,Neu5,chi Ac2), and poly(Neu5Gc, Neu5-Gc chi Ac), where chi represents the site of acetylation at carbon atom 4, 7, or 9. The significance of this new structural information, together with our recent finding of alpha 2-->8 linked polydeaminoneuraminic acid, poly(KDN), in the rainbow trout egg vitelline envelope, is that it demonstrates the natural occurrence of multiple forms of alpha 2-->8-linked polySia chains in Salmonidae fish glycoproteins that have not been previously described. The results also predict that a remarkable array of polysialylated glycoconjugates is yet to be discovered in animals other than teleost fishes. PMID- 8226895 TI - Characterization of the structural requirements for assembly and nucleotide binding of an ATP-binding cassette transporter. The maltose transport system of Escherichia coli. AB - The periplasmic maltose-binding protein-dependent, maltose transport system of Escherichia coli is a well studied member of the ATP-binding cassette family of transport ATPases. In addition to the water-soluble maltose-binding protein, the system comprises three membrane proteins, MalF, MalG, and MalK, which form a heterotetrameric complex (FGK2) in the cytoplasmic membrane. The purified complex exhibits transport-associated ATPase activity. To characterize the requirements for nucleotide binding and hydrolysis by the FGK2 complex, we used plasmids to express different combinations of the individual subunits as well as mutant forms of the MalK subunit. Prior to measuring nucleotide binding, we examined membrane preparations for the presence of each subunit from strains that contained all possible permutations of the three structural genes, malF, malG, and malK. We found that when all three genes were present or when malF and malK were present together, the corresponding antigens were detected easily on Western immunoblots and were soluble in the non-ionic detergent, Triton X-100. In contrast, all other permutations resulted in decreased amounts of antigen or antigen that was Triton X-100-insoluble. We relied on photocross-linking with 8-azido-[32P]ATP and ATP hydrolysis as indicators of the ability of the transport complex to interact with purine nucleotides. 8-Azido-[32P]ATP was photocross-linked to the MalK subunit. Photolabeling of MalK was inhibited by ATP, ADP, and GTP and not by other nucleotides. Photolabeling of MalK required the presence of MalF but not MalG. Mutations in malK that affect amino acid residues thought to be directly involved in nucleotide binding did indeed abolish labeling and resulted in loss of transport activity without affecting protein stability. In general, ATP hydrolysis correlated with the photocross-linking. A notable exception is the MalK941 mutant protein which retained the ability to be labeled by 8-azido [32P]ATP but was unable to catalyze detectable levels of ATP hydrolysis. Some, but not all, of the malK mutations were dominant to wild type. To study the mechanism of dominance we devised a means of measuring the ability of different wild-type and mutant MalK proteins to interact with the MalF and MalG subunits. This assay relies on the fact that, when a bifunctional MalK-LacZ hybrid protein is associated with the MalF and MalG subunits, it is membrane-bound. Excess MalK competed with the MalK-LacZ hybrid protein for sites in the membrane and resulted in the hybrid fractionating as a soluble protein.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8226896 TI - Transcytosis of placental alkaline phosphatase-polymeric immunoglobulin receptor fusion proteins is regulated by mutations of Ser664. AB - Two signals are known that regulate the entry of the polymeric immunoglobulin receptor (pIgR) into the transcytotic pathway: binding of the ligand, dimeric IgA, and phosphorylation of Ser664 in the cytoplasmic domain of the receptor. Mutation of Ser664 to an alanine residue generates a receptor that is transcytosed slowly and exhibits increased recycling at the basolateral plasma membrane. In contrast, if Ser664 is mutated to an aspartate (which bears a constant negative charge-like a phosphate group) the receptor is transcytosed at a rate that is greater than the wild-type pIgR. We have constructed fusions of placental alkaline phosphatase (PLAP) with the transmembrane region and cytoplasmic domain of the wild-type or mutant pIgRs described above (PLAP/WT, PLAP/Ala, and PLAP/Asp) in order to study transcytotic signals present in the cytoplasmic domain of the pIgR independently of those present in the lumenal domain. Transcytosis of the fusion proteins was measured by cell surface immunoprecipitation of a single cohort of basolaterally biotinylated fusion proteins as it arrived at the apical plasma membrane. Significantly more of the PLAP/WT fusion (27%) than the PLAP/Ala fusion protein (5%) was transcytosed during a 2-h chase period. In addition, it was found that approximately 10% of a pseudoligand, monovalent 125I-labeled Fabs (derived from affinity-purified anti PLAP antibodies), was transcyosed by the PLAP/WT fusion, 7% by the PLAP/Ala fusion, and 22% by the PLAP/Asp fusion over a 4-h period. These data are consistent with the hypothesis that the cytoplasmic domain of the pIgR contains information necessary to direct proteins into the transcytotic pathway, albeit inefficiently, and that phosphorylation of Ser664 is at least a partially autonomous signal for transcytosis. PMID- 8226897 TI - In vivo and in vitro phosphorylation of the T lymphocyte type n (Kv1.3) potassium channel. AB - The major species of voltage-gated potassium channel found on mammalian T lymphocytes is referred to as the type n channel. This potassium channel exhibits unique functional properties which distinguish it from other species of potassium channels, including a potential role in the onset of cellular events associated with T cell activation. As a first step in characterizing specific biochemical properties of the type n channel, we have generated polyclonal antisera against bacterial fusion proteins containing peptide regions unique to the mouse and human type n channel. From membranes of T cell lines derived from both mouse (SAK 8 cell line) and human (Jurkat cell line), the type n channel can be immunoprecipitated following either surface labeling with 125I or metabolic labeling with 32P. The apparent molecular mass of the immunoprecipitated type n channel is approximately 65 kDa, significantly greater than that of the 58-kDa in vitro translated product, and suggestive of post-translational modification events. Phosphoamino acid analysis of the metabolically labeled Jurkat type n channel reveals phosphorylation of serine residues exclusively. In vitro studies also describe the ability of both protein kinase A and protein kinase C to phosphorylate the Jurkat type n channel. The former kinase also appears to phosphorylate a 40-kDa protein which co-immunoprecipitates with the type n channel. These data suggest that direct phosphorylation of the T lymphocyte type n potassium channel or its associated 40-kDa subunit may serve as a means by which channel activity is regulated. PMID- 8226898 TI - The highest levels of purine catabolic enzymes in mice are present in the proximal small intestine. AB - Recent studies on the tissue distribution and developmental regulation of adenosine deaminase (ADA) activity in mice show that very high ADA levels exist in the murine alimentary tract (tongue, esophagus, forestomach, proximal small intestine) and at the fetal-maternal interface. To understand the role of ADA in these tissues, we measured the levels of three other enzymes involved in purine catabolism, purine nucleoside phosphorylase (PNP), guanine deaminase (GDA), and xanthine dehydrogenase (XDH), to see how their levels correlated with ADA activity. Our results show that the highest level of PNP, GDA, and XDH is present in the proximal small intestine. Levels of these purine catabolic enzymes are much lower in the tongue, esophagus, forestomach, and fetal-maternal interface in marked contrast to ADA distribution. We also determined mRNA levels encoding PNP, XDH, and ADA in a variety of tissues. Tissue-specific differences in PNP, XDH, and ADA activity correlated with RNA abundance, indicating that the regulation of gene expression is at the level of mRNA production. Thus, ADA is part of a purine catabolic pathway leading to the production of uric acid that is present at the highest known level in the proximal small intestine. ADA may have additional roles in other tissues. PMID- 8226899 TI - Structure and mechanism of the G protein-coupled receptor kinases. PMID- 8226900 TI - Primary structure and muscle-specific expression of the 50-kDa dystrophin associated glycoprotein (adhalin). AB - The 50-kDa dystrophin-associated glycoprotein (50-DAG) is a component of the dystrophin-glycoprotein complex, which links the muscle cytoskeleton to the extracellular matrix. 50-DAG is specifically deficient in skeletal muscle of patients with severe childhood autosomal recessive muscular dystrophy and in skeletal and cardiac muscles of BIO 14.6 cardiomyopathic hamsters. The lack of 50 DAG leads to a disruption and dysfunction of the dystrophin-glycoprotein complex in these diseases. The cDNA encoding 50-DAG has now been cloned from rabbit skeletal muscle. The 50-DAG deduced amino acid sequence predicts a novel protein having 387 amino acids, a 17-amino acid signal sequence, one transmembrane domain, and two potential sites of N-linked glycosylation. Affinity-purified antibodies against rabbit 50-DAG fusion proteins or synthetic peptides specifically recognized a 50-kDa protein in skeletal muscle sarcolemma and the 50 kDa component of the dystrophin-glycoprotein complex. In contrast to dystroglycan, which is expressed in a wide variety of muscle and non-muscle tissues, 50-DAG is expressed only in skeletal and cardiac muscles and in selected smooth muscles. Finally, 50-DAG mRNA is present in mdx and Duchenne muscular dystrophy (DMD) muscle, indicating that the down-regulation of this protein in DMD and the mdx mouse is likely a post-translational event. PMID- 8226901 TI - Mas6p can be cross-linked to an arrested precursor and interacts with other proteins during mitochondrial protein import. AB - Mas6p is an integral membrane protein of the yeast mitochondrial inner membrane, which is essential for mitochondrial protein import (1). To determine whether Mas6p is directly involved in recognizing precursors or translocating them across the inner membrane, we asked if Mas6p was in close proximity to precursor proteins being imported into mitochondria. We report here that Mas6p can be chemically cross-linked to an imported protein arrested in transit through the mitochondrial inner membrane. Antiserum to Mas6p specifically immunoprecipitates one of several different mitochondrial proteins that are cross-linked to blocked precursors. Our results strongly suggest that Mas6p physically interacts with precursors during their translocation into the matrix. In addition, at least two other mitochondrial proteins that are each cross-linked to arrested precursors can be coimmunoprecipitated along with Mas6p under non-denaturing conditions. These observations provide evidence for a complex of proteins including Mas6p, each of which interacts with mitochondrial precursors during import. PMID- 8226902 TI - Expression of human apolipoprotein B100 in transgenic mice. Editing of human apolipoprotein B100 mRNA. AB - Apolipoprotein B (apoB) is a large glycoprotein that circulates in plasma as a major constituent of numerous lipoproteins. ApoB exists in two forms: apoB48 and apoB100. ApoB48 is identical in sequence to the N-terminal region of apoB100 and is generated by sequence-specific mRNA editing of the apoB100 transcript. Here, we describe the development of a line of mice expressing a human apoB transgene driven by promoter/enhancer sequences from the transthyretin gene. In these mice, immunodetectable human apoB100 is synthesized by the liver, kidney, and brain. Human apoB100 is found in low concentration (approximately 0.1 mg/dl) in the plasma of the transgenic mice and circulates in the low density lipoprotein fraction. The hepatic human apoB100 transcripts undergo mRNA editing at only slightly lower efficiency than the endogenous mouse apoB100 message. Therefore, there is no absolute species specificity to the apoB100 mRNA editing process. PMID- 8226903 TI - A matrix ATP requirement for presequence translocation across the inner membrane of mitochondria. AB - The mitochondrial presequence initiates protein translocation across the inner membrane of mitochondria in a delta psi-dependent step. We have investigated the role of matrix ATP in this process. When matrix ATP was reduced to interfere with the function of mitochondrial heat shock protein 70, presequence translocation across the inner membrane was strongly inhibited. This was accompanied by the accumulation of an import intermediate that was unprocessed and accessible to protease added to the intact mitochondria. Both delta psi and matrix ATP were required for further translocation of this intermediate into the matrix. When ATP levels are insufficient to support protein import, it appears that the presequence becomes translocated across the inner membrane, but delta psi does not maintain it in the matrix. Presequence translocation across the inner membrane is thus a reversible reaction, and a step dependent on matrix ATP is required to make it unidirectional. Based on these observations, a model on the role of delta psi, mthsp70, and matrix ATP in presequence translocation across the inner membrane is presented. PMID- 8226904 TI - UV damage to DNA strongly influences its rotational setting on the histone surface of reconstituted nucleosomes. AB - The major photoproduct formed in DNA, following absorption of ultraviolet (UV) light, is the cis-syn cyclobutane pyrimidine dimer (CPD). Formation of CPDs in DNA packaged into chromatin prior to UV irradiation results in a striking preference for these photoproducts to be oriented away from the histone surface in nucleosomes (Gale, J. M., Nissen, K. A., and Smerdon, M. J. (1987) Proc. Natl. Acad. Sci. U. S. A. 84, 6644-6648). In this report, we show that formation of nucleosomes onto UV-irradiated DNA results in a similar distribution of these photoproducts, indicating that preexisting CPDs in the DNA molecule can influence its rotational setting on the histone surface during nucleosome formation. This bias is less pronounced in the central three helical turns encompassing the dyad axis of nucleosomes, where the helix is overwound (10.7 bases/turn) relative to the "outer" portion of core DNA (10.0 bases/turn). Such a change in the rotational setting of DNA on the surface of newly formed nucleosomes could expose normally inaccessible DNA sequences to factors which control DNA processing. PMID- 8226905 TI - A protein cofactor is required for uncoating of clathrin baskets by uncoating ATPase. AB - Immediately after clathrin-coated pits pinch off from the cell membrane to form clathrin-coated vesicles, clathrin dissociates from the vesicles. In vitro studies suggest that this dissociation is carried out by the uncoating ATPase, a constitutive member of the 70-kDa heat shock family. Aside from the requirement for ATP, nothing is known about the regulation of the uncoating process. We now show that clathrin baskets prepared from highly purified clathrin and AP2, the assembly protein associated with plasma membrane coated vesicles, cannot be uncoated by the bovine brain uncoating ATPase alone. A 100-kDa protein cofactor, which was isolated from coated vesicles, is essential for uncoating by the uncoating ATPase. This cofactor restores normal uncoating when present at a molar ratio of about 1 to 10 to clathrin and uncoating ATPase. PMID- 8226906 TI - Base selection, proofreading, and mismatch repair during DNA replication in Escherichia coli. AB - The accuracy by which organisms duplicate their DNA is of considerable interest. At least three mechanisms operate, serially, to secure high fidelity: base selection, exonucleolytic proofreading, and postreplicative mismatch correction. To obtain insights into the efficiency and specificity of these steps in the bacterium Escherichia coli, we have performed DNA sequence analysis of mutations occurring in the bacterial lacI gene in a series of strains genetically disabled in one or more of these error avoidance pathways. The base selection efficiency was estimated from mutagenesis occurring in a mutDmutL strain, which is deficient in both proofreading (mutD5) and mismatch repair (mutL). The proofreading efficiency was derived comparing the mutD5 mutL strain to the mismatch repair deficient mutL strain. The efficiency of mismatch repair was derived comparing the mutL strain to the wild-type strain. The results show that base selection discriminates against errors by 200,000-2,000,000-fold, proofreading by 40-200 fold, and mismatch repair by 20-400-fold, each depending on the type of error. Base selection and proofreading act more strongly against transversions than transitions, whereas mismatch repair does the opposite. The data are based on 866 sequenced lacI mutations in a target that allows the scoring of at least 127 different mutations in 76 distinct DNA sequence contexts in vivo. They may therefore have general significance. PMID- 8226907 TI - Modification of the calcium and calmodulin sensitivity of the type I adenylyl cyclase by mutagenesis of its calmodulin binding domain. AB - The type I adenylyl cyclase is directly stimulated by Ca2+ and calmodulin in vitro, and the enzyme is also stimulated by increases in intracellular Ca2+ in vivo. Ca2+ stimulation of the enzyme in vivo may be due to direct interactions of the enzyme with Ca2+ and calmodulin or to an indirect mechanism involving stimulation of the enzyme by Ca(2+)-activated protein kinases. In this study, we have made several point mutations within the calmodulin binding domain to determine if the Ca2+ sensitivity of the enzyme can be modified by mutagenesis. The catalytic activities of the mutant enzymes were comparable to wild type type I adenylyl cyclase. Substitution of Cys-507 with Ser-507 did not have significant effects on the calmodulin or Ca2+ sensitivity of the enzyme. However, replacement of Lys-504 with Asp caused a 4-fold decrease in sensitivity to Ca2+. Ca2+ and calmodulin stimulation were abolished by substitution of Phe-503 with Arg-503. Stimulation of type I adenylyl cyclase activity in vivo by intracellular Ca2+ was also greatly diminished with the Arg-503 mutant indicating that Ca2+ stimulation of the enzyme in vivo is due primarily to direct interactions with calmodulin and Ca2+. These data demonstrate that the Ca2+ sensitivity of this enzyme can be modulated by point mutagenesis within the putative calmodulin binding domain and indicate that the enzyme can be directly regulated by Ca2+ and calmodulin in vivo. PMID- 8226908 TI - Increased palmitoylation of the Gs protein alpha subunit after activation by the beta-adrenergic receptor or cholera toxin. AB - The alpha subunit of the heterotrimeric Gs protein that couples the beta adrenergic receptor to adenylyl cyclase undergoes post-translational palmitoylation. We examined the dynamics of this modification of alpha s by metabolic labeling of COS and S49 lymphoma cells under different conditions. The endogenous alpha s proteins were immunoprecipitated with a peptide-specific antibody, separated by SDS-polyacrylamide gel electrophoresis, and analyzed by fluorography and densitometry. A pulse-chase study of COS cells incubated with [3H]palmitate or [35S]methionine showed that for alpha s the palmitate turnover (t1/2 approximately 50 min) was significantly faster than the protein degradation. Treatment of cells with 10 microM isoproterenol, a beta-adrenergic receptor agonist, in the presence of [3H]palmitate led to a rapid 4-10-fold increase in the palmitoylation of alpha s. This increase in palmitoylation was concentration-dependent (EC50 approximately 0.9 microM) and blocked by the antagonist propranolol. The mutant alpha s proteins in the unc and H21a S49 cell lines did not show an increase in [3H]palmitate incorporation with isoproterenol treatment. Cholera toxin treatment of COS cells increased the [3H]palmitate incorporation into the alpha s subunits. These data indicate that palmitoylation of the alpha s subunit is dynamic and regulated by activation of the alpha s subunit. PMID- 8226909 TI - Prenylation of Rab5 is dependent on guanine nucleotide binding. AB - Rab5 is a small molecular weight GTP-binding protein that functions in endocytic vesicle traffic. Like other Ras-related proteins, Rab5 is prenylated on C terminal cysteine residues, although it lacks the typical C-terminal CAAX motif (where A is any aliphatic amino acid and X is any amino acid) to direct this post translational modification. We have investigated structural requirements for the in vitro geranylgeranylation of Rab5. Rab5N133I, a point mutant that has impaired ability to bind GTP or GDP, undergoes modification to a limited extent and at a severely reduced rate when compared to cognate Rab5. A second point mutant, Rab5Q79L, can be processed to approximately the same extent as wild-type albeit at a reduced rate. Since the latter mutation results in defective GTPase activity, these combined observations indicate that guanine nucleotide binding plays an important role in the geranylgeranylation reaction and suggest that the GDP-bound form of Rab5 is the preferred conformation for interaction with Rab prenyltransferase. This idea is supported by the finding that non-hydrolyzable GTP analogs inhibit Rab5 prenylation, while in vitro processing of both H-ras and the gamma 2 subunit of regulatory G proteins is unaffected at concentrations of guanosine 5'-O-(thiotriphosphate) (GTP gamma S) up to 400 microM. Moreover, a truncation mutant lacking the C-terminal cysteines, Rab5(1-211), serves as an inhibitor of Rab5wt geranylgeranylation when liganded with GDP but not GTP gamma S. Thus, the recognition of Rab5 as a substrate by Rab prenyltransferase involves structural elements exclusive of the C terminus and dependent upon the GDP binding conformation of the protein. PMID- 8226910 TI - Pursuing the voltage sensor of a voltage-gated mammalian potassium channel. AB - Ion channels are generally classified as voltage- or ligand-gated channels based upon their ability to respond to a change in the transmembrane voltage or to a specific ligand interaction. The voltage sensor of voltage-gated ion channels is thought to be contained within the fourth putative transmembrane segment, S4, and is characterized by basic residues typically spaced apart. Thus far, a full understanding of all charges contributing to the voltage sensor of voltage-gated potassium channels has been lacking since several neutralization mutants in S4 could not be functionally expressed. By construction of multimeric cDNAs encoding a mammalian RCK1 (Kv1.1) potassium channel, we now report functional expression of all charge neutralizations in the S4 segment, providing a more comprehensive insight into the functioning of the voltage sensor. PMID- 8226911 TI - Evidence for the covalent linkage of carbohydrate polymers to a glycoprotein from Streptococcus sanguis. AB - The platelet aggregation-associated protein (PAAP) from Streptococcus sanguis contains 39% carbohydrate in rhamnose-rich polysaccharides. Synthesized by cultured protoplasts, these polysaccharides were shown to be covalently linked to this cell wall protein using specific inhibitors of glycosylation, glycosidase treatment, amino acid and carbohydrate analyses of prepared minimal glycopeptides and isolated protein, and NMR spectroscopy. To our knowledge, this is the first direct proof of an N-asparaginyl linkage of carbohydrate to a eubacterial protein. PMID- 8226912 TI - Identification of the nerve terminal targets of botulinum neurotoxin serotypes A, D, and E. AB - Botulinum neurotoxins are metalloproteins with one zinc atom bound to the zinc binding motif of zinc endopeptidases. Here we show that botulinum neurotoxin serotypes A, D, and E are zinc endoproteases specific for components of the synaptic vesicle docking and fusion complex. Serotypes A and E cleave SNAP-25, a 25-kDa protein of the synaptic terminal, while serotype D is specific for VAMP/synaptobrevin, a membrane protein of synaptic vesicles. Both rat brain VAMP isoforms are cleaved at a single Lys-Leu peptide bond. The proteolytic activity of these neurotoxins is inhibited by EDTA and captopril. PMID- 8226913 TI - Activation of calmodulin-dependent enzymes can be selectively inhibited by histone H1. AB - Calmodulin (CaM) and its target enzymes are important regulators of a variety of cellular processes including gene expression and cell cycle progression (Bading, H., Ginty, D. D., and Greenberg, M. E. (1993) Science 260, 181-186; Rasmussen, C. D., and Means, A. R. (1989) EMBO J. 8, 73-82). It has been previously accepted that regulation of CaM-dependent enzyme activity occurs via calcium/calmodulin dependent activation. We have found that histone H1 is a potent inhibitor of the CaM-dependent activation of mouse calcium/calmodulin-dependent protein kinase II (CaMKII) and of the CaM-dependent protein phosphatase, calcineurin. Inhibition is mediated only by free histone H1; addition of DNA abolishes the inhibitory effect. The effect is not due to a simple interaction of basic (histone) and acidic (CaM) proteins since myelin basic protein and histone H2B, CaMKII substrates more basic than histone H1, did not affect autophosphorylation of CaMKII, and myelin basic protein had no effect on calcineurin activity. The effect is specific to CaM since addition of parvalbumin, a related Ca(2+)-binding protein, did not reverse the effect of histone H1, whereas addition of CaM recovered enzyme activity. These results indicate that free histone H1 levels may specifically affect the ability of CaM to activate its target enzymes and suggests a novel level of control of CaM-dependent enzymes in eukaryotic cells. PMID- 8226914 TI - Single-stranded DNA binding activity of C1-tetrahydrofolate synthase enzymes. AB - In eukaryotes C1-5,6,7,8-tetrahydrofolate (THF) synthase is a trifunctional enzyme that catalyzes the interconversion of reduced forms of folate to supply activated one-carbon units required for a variety of metabolic pathways. The enzymatic activities include 10-formyl-THF synthetase (EC 6.3.4.3), 5,10-methenyl THF cyclohydrolase (EC 3.5.4.9), and 5,10-methylene-THF dehydrogenase (EC 1.5.1.5). In bacteria separate, monofunctional or bifunctional polypeptides catalyze the same reactions. We have purified C1-THF synthase from the fission yeast Schizosaccharomyces pombe and found its physical and enzymatic properties similar to those of other eukaryotic C1-THF synthase enzymes. Unexpectedly, the S. pombe enzyme bound strongly (Keq = 100 pM) to single-stranded DNA, but not to double-stranded DNA or to RNA. The binding was sequence-independent, apparently not cooperative, and not detectably inhibited by C1-THF synthase substrates or cofactors. Trifunctional cytoplasmic enzyme from Saccharomyces cerevisiae and monofunctional (synthetase) enzyme from Clostridium acidiurici also bound tightly to single-stranded DNA, while bifunctional (dehydrogenase and cyclohydrolase) enzyme from Escherichia coli did not, suggesting that single-stranded DNA binding is a conserved function of the synthetase domain of C1-THF synthase enzymes. PMID- 8226915 TI - Control of yeast glycogen synthase-2 by COOH-terminal phosphorylation. AB - The budding yeast Saccharomyces cerevisiae expresses two isoforms of glycogen synthase, of which glycogen synthase-2 (GS-2) appears to be the most important determinant of glycogen accumulation (Farkas, I., Hardy, T. A., Goebl, M. G., and Roach, P. J. (1991) J. Biol. Chem. 266, 15602-15607). Partial proteolysis of purified yeast glycogen synthase activated the enzyme, mimicking the effects of dephosphorylation. The cleavage was localized to the COOH terminus of the molecule and trypsin treatment released 32P from enzyme labeled in vivo with 32P or in vitro by cyclic AMP-dependent protein kinase. Similarly, when cells were labeled with 32P, no radioactivity was incorporated into a mutant form of GS-2 truncated at residue 643 while the wild type enzyme was phosphorylated at both Ser and Thr residues. The 9 Ser and Thr residues COOH-terminal to position 643 were mutated individually to Ala, and the GS-2 mutants were expressed from a low copy plasmid in yeast that lacked functional chromosomal copies of the two glycogen synthase genes. Mutations at Ser-650, Ser-654, and Thr-667 resulted in significant activation of yeast glycogen synthase and elevation in the level of accumulated glycogen as compared with wild type. Likewise, expression of the truncated GS-2 resulted in hyperactive enzyme and the overaccumulation of glycogen. None of the other Ser or Thr mutations substantially affected glycogen synthase activity and glycogen storage. We conclude that Ser-650, Ser-654, and Thr-667 are regulatory phosphorylation sites in vivo. However, in vitro, cyclic AMP-dependent protein kinase modified Ser residue(s) COOH-terminal to position 659, and so the identity of the physiological GS-2 kinases is unclear. Yeast strains bearing glc7 and gac1 mutations are defective in genes encoding type 1 protein phosphatase components and are impaired in their ability to accumulate glycogen. Expression of the truncated GS-2 in these strains restored glycogen accumulation, as did the presence of GS-2 mutated at Ser-650, Ser-654, or Thr 667. These data are consistent with the hypothesis that type 1 phosphatase regulates GS-2 by controlling its phosphorylation state. PMID- 8226916 TI - Polymerase processivity and termination on Drosophila heat shock genes. AB - 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) limits RNA polymerase II transcription to a gene's 5'-end. Transcription of the uninduced Drosophila hsp70 gene is likewise restricted to the 5'-end, where the polymerase resides in a paused state. Furthermore, paused elongation complexes formed on the uninduced hsp70 gene and in DRB-inhibited reactions can both be restarted by Sarkosyl or high salt. These similarities prompted us to explore whether these complexes were generated by a block at the same polymerase modification step. In vivo UV cross linking and KMnO4 hyperreactive site mapping show that while the naturally paused polymerase is restricted to the first approximately 42 base pairs of hsp70, DRB treatment of heat-induced cells allows the polymerase to transcribe past this site. Therefore, the DRB-sensitive step is probably not rate-limiting for hsp70 transcription under uninduced conditions. DRB treatment did, however, lead to the reduction of KMnO4 hyper-reactivity on hsp70 and hsp26 in a region correlating with open polymerase and/or early elongation complexes, suggesting a site for the DRB-sensitive polymerase modification step. Finally, we used the techniques of polymerase-DNA cross-linking and KMnO4 hyper-reactive site mapping to analyze the natural polymerase termination process at the 3'-end of the hsp26 gene. The data obtained are consistent with polymerases terminating at multiple sites downstream of the polyadenylation site. PMID- 8226917 TI - Differential recognition of OR1 and OR3 by bacteriophage 434 repressor and Cro. AB - The developmental decisions of bacteriophage 434 depend on the ability of 434 repressor and Cro to bind OR1 and OR3 with different relative affinities; repressor binds OR1 tighter than OR3, whereas Cro slightly prefers OR3 over OR1. Studies with operator mutants show that repressor's lower relative affinity for OR3 results from a deviation in the sequence of OR3 from consensus; an A-->G change at position 4 in one half-site (OR1: A-C-A-A-A-C-T-T-T-C-T-T-G-T; OR3: A-C A-G-T-T-T-T-T-C-T-T-G-T). Similar experiments show that Cro binds operators containing either A.T or G.C bases pairs at position 4 equally well, but cannot bind operators containing C.G or T.A base pairs at this position. A Gln33-->Ala mutation in 434 repressor diminishes, but does not eliminate, its ability to distinguish between purines at position 4. This shows that a glutamine at amino acid 33 is not the sole determinant of repressor's position 4 specificity. Changing Gln33-->Leu, the amino acid at the homologous position in Cro, does not confer "Cro-like" position 4 base specificity on repressor. Similarly, a Cro protein bearing Gln at this position does not exhibit repressor's position 4 base preferences. The residual specificities of these mutant proteins indicates that in each protein, more than 1 amino acid is responsible for recognizing bases at position 4. These were identified by analyzing the binding specificities of multiply mutated repressors, in vitro. The types of substitutions made were guided by sequence homologies between 434 repressor and Cro. At least three mutations are needed to eliminate repressor's position 4 base specificity; Gln33- >Ala, Glu32-->Gln, and Thr27-->Lys, although no set of amino acid substitutions in repressor was able to confer Cro-like position 4 specificity to repressor. These results indicate that at least the amino acids at these positions are involved in recognition of the position 4 base. Other evidence suggests that Cro and repressor use identical amino acids present at homologous positions in the DNA recognition helix in different ways. PMID- 8226918 TI - Betaine-aldehyde dehydrogenase from leaves of Amaranthus hypochondriacus L. exhibits an Iso Ordered Bi Bi steady state mechanism. AB - The kinetics of the oxidation of betaine aldehyde catalyzed by NAD(+)-betaine aldehyde dehydrogenase, purified from amaranth leaves subjected to water deficit, were analyzed by steady state initial velocity and product and dead-end inhibition studies at low substrate concentrations. Only one product, NADH, gives inhibition. The other product of the reaction, glycine betaine, does not inhibit the enzyme even at concentrations as high as 10 mM. In dead-end inhibition experiments, AMP and choline were used as dead-end analogs of NAD+ and betaine aldehyde, respectively. The families of double-reciprocal plots in the range 0.010-0.500 mM NAD+ and 0.025-0.300 mM betaine aldehyde are linear and intersect at the left of the 1/v axis. NADH is a mixed inhibitor against NAD+ and betaine aldehyde. AMP is competitive with respect to NAD+ and mixed with betaine aldehyde. Choline is competitive against betaine aldehyde and uncompetitive with respect to NAD+. Our results are consistent with an Iso Ordered Bi Bi steady state mechanism in which NAD+ is the first substrate to bind to the enzyme and NADH is the last product to dissociate from it. To our knowledge, this is the first time that an Iso mechanism has been demonstrated by product inhibition studies, as predicted by Cleland (Cleland, W. W. (1963) Biochim. Biophys. Acta 67, 104-137). PMID- 8226919 TI - Cloning and characterization of a unique elastolytic metalloproteinase produced by human alveolar macrophages. AB - Human alveolar macrophages have the capacity to degrade elastin. As an approach to define proteinases responsible for this activity, we recently cloned a murine macrophage elastase cDNA and demonstrated that it is a member of the matrix metalloproteinase gene family (Shapiro, S. D., Griffin, G. L., Gilbert, D. J., Jenkins, N. A., Copeland, N. G., Welgus, H. G., Senior, R. M., and Ley, T. J. (1992) J. Biol. Chem. 267, 4664-4671). We now report that there is a human orthologue of murine macrophage metalloelastase that we call human macrophage metalloelastase (HME). The full-length HME cDNA spans 1.8 kilobases and contains an open reading frame of 1410 base pairs; the predicted molecular mass of the HME proenzyme is 54 kDa. HME mRNA and protein were detected in human alveolar macrophages. Similar to murine macrophage metalloelastase, HME readily undergoes NH2- and COOH-terminal processing to a mature 22-kDa form. Both recombinant HME expressed in Escherichia coli and native HME derived from human alveolar macrophage-conditioned media degraded insoluble elastin. HME is a unique human metalloproteinase that possesses elastolytic activity and is expressed in alveolar macrophages; it is therefore a candidate molecule for the causation of diseases characterized by damage to the extracellular matrix. PMID- 8226920 TI - The mechanism of sequence-specific DNA cleavage and strand transfer by phi X174 gene A* protein. AB - We have examined the biological role and catalytic function of two juxtaposed tyrosyl residues in the bacteriophage phi X174 gene A protein, Tyr-343 and Tyr 347, which have been implicated in the catalysis of sequence-specific DNA strand transfer. Site-directed mutagenesis changing either tyrosine to phenylalanine abolishes phage viability. The biochemical basis of this inviability was studied using purified A* protein containing the carboxyl-terminal 341 amino acids of the A protein, as well as purified A* protein with a Y343F or Y347F mutation. All three proteins can cleave the phi X174 replication origin and perform strand transfer between oligodeoxynucleotides bearing the recognition sequence of the A protein; however, both Tyr-343 and Tyr-347 appear to be required for coordinated DNA strand transfer by a single A* protein molecule. The chirality of a phosphorothioate group at the site of strand transfer in the DNA was found to be retained following the strand-transfer reaction, which argues against transfer of Tyr-343-linked DNA to Tyr-347 on the same protein molecule or vice versa. These results support the current model of gene A protein function in which the two tyrosines of a single protein molecule alternate in catalyzing DNA strand transfer at the viral replication origin. PMID- 8226921 TI - Role of the conserved Lys-X-Gly-Gly sequence at the ADP-glucose-binding site in Escherichia coli glycogen synthase. AB - Although bacterial and mammalian glycogen synthases differ in the primary structure and specificity for glucosyl donor, lysyl residues identified at their substrate-binding sites by affinity labeling are present in a conserved tetrapeptide sequence, Lys-X-Gly-Gly, where X is a residue not conserved (Tagaya, M., Nakano, K., and Fukui, T. (1985) J. Biol. Chem. 260, 6670-6676; Furukawa, K., Tagaya, M., Inouye, M., Preiss, J., and Fukui, T. (1990) J. Biol. Chem. 265, 2086 2090). To elucidate the functional role of this conserved sequence, Lys-15, Gly 17, and Gly-18 in Escherichia coli glycogen synthase have been replaced by other amino acid residues via site-directed mutagenesis. Kinetic analyses of the Lys-15 mutant enzymes showed that the epsilon-amino group of Lys-15 is mainly involved in binding of the phosphate moiety adjacent to the glycosidic linkage in the substrate ADP-glucose, presumably through an ionic interaction. The mutant enzyme in which Ala was substituted for Gly-17 had a catalytic rate constant 3 orders of magnitude smaller than that of the wild-type enzyme with a slightly increased Michaelis constant for ADP-glucose, whereas the Gly-18-->Ala mutant showed a rate constant only 3.2-fold smaller. In addition, mutations of Gly-17 and Gly-18 resulted in marked changes in the reactivity of Lys-15 with affinity labeling reagents. These results suggest that the 2 glycyl residues in the conserved Lys-X Gly-Gly sequence, in particular the one closer to the ADP-glucose-binding lysyl residue, participate in catalysis by assisting conformational change(s) of the active site or stabilizing the transition state. PMID- 8226922 TI - Phosphatidic acid and diacylglycerol synergize in a cell-free system for activation of NADPH oxidase from human neutrophils. AB - NADPH oxidase, the respiratory burst enzyme of human neutrophils, is a multi component complex that is assembled and activated during stimulation of the cells by inflammatory or phagocytic stimuli. The signal mechanisms leading to activation of the enzyme are unclear, but it is likely that phospholipases are involved. Recent work has shown that phosphatidic acid, the initial product of phospholipase D activation, is a weak activator of NADPH oxidase in a cell-free system. We now show that diacylglycerol enhances the cell-free activation of NADPH oxidase activation by phosphatidic acid. 1,2-Didecanoyl phosphatidic acid (10:0-PA) and 1,2-dioctanoylglycerol (8:0-DG) each increased levels of NADPH oxidase activity in mixtures of membrane and cytosolic fractions about 2-fold. The combination of both lipids increased NADPH oxidase activity approximately 12 fold, indicative of a synergistic response. Fatty acid and neutral lipid metabolites of 10:0-PA or 8:0-DG were ineffective, suggesting activation is directly mediated by phosphatidic acid and diacylglycerol. Activation was time- and concentration-dependent with maximum activation at 30-60 min and a sharp peak of maximal activity at 10 microM 10:0-PA and 30 microM 8:0-DG. In lipid specificity studies, activity of PA or DG decreased with increasing acyl chain length but was restored by introducing unsaturation in the acyl chain. Natural forms of PA stimulated levels of activity comparable to that seen with 10:0-PA. Synthetic and natural phosphatidylserines, but not other phospholipids, could replace phosphatidic acid in the synergistic response. These studies provide direct evidence for a synergistic interaction between phosphatidic acid and diacylglycerol in mediating a cellular function: the assembly and activation of NADPH oxidase. Our results support the concept that the generation of second messenger lipids by phospholipase D is a key step in activation of the respiratory burst enzyme. PMID- 8226923 TI - Correlation of myosin light chain phosphorylation with isometric contraction of fibroblasts. AB - In vitro studies have indicated that the enzymatic activity of myosin II from non muscle cells is controlled by phosphorylation of its regulatory light chain (LC20). We have studied one likely functional consequence of phosphorylating LC20 in living chick embryo fibroblasts (CEF) by measuring contractile force developed by these cells. Using a recently developed method, we recorded quantitative changes in isometric force generated by a population of cells following mitogenic stimulation. Fetal bovine serum, thrombin, and lysophosphatidic acid stimulate rapid isometric contraction of CEF. Cells stimulated with thrombin develop maximal force within 5-10 min. Force development correlates temporally with a 3-5 fold increase in the overall fraction of LC20 phosphorylated and with the fractions of LC20 in both the monophosphorylated and diphosphorylated states. Unloaded shortening velocity also increases after thrombin stimulation. Although both force and phosphorylation begin to decline 10 min after stimulation, the level of phosphorylation declined more rapidly than the force. These results suggest that the role of LC20 phosphorylation in regulating fibroblast contractility is analogous to its well established role in regulating smooth muscle contraction and that quantitative measurements of the force developed by populations of fibroblasts (or other cultured cells) can be used to study the regulation of non-sarcomeric myosin at the molecular level in vivo. PMID- 8226924 TI - Selective release of peptides from lysosomes. AB - We demonstrate a selective release of peptides by lysosomes in vitro. A lysosomal fraction from human fibroblasts that had previously endocytosed [3H]ribonuclease A was incubated for 2 h, and radioactivity released into the medium and radioactivity retained within lysosomes were analyzed. A variety of radiolabeled molecules including peptides of an appropriate size to serve as antigens for T cell-mediated immunity were released. One small peptide was predominantly released, while others, as well as intact ribonuclease A, were predominantly retained. A 4-5-fold range was also evident in the relative release of three 3H labeled tripeptide probes of similar charge derived from the sequence of ribonuclease A. This selectivity and the fact that similar peptide degradation fragments were also released and retained by intact cells after endocytosis of [3H]ribonuclease A argues strongly that the release observed in vitro is physiological and not due to damaged lysosomal membranes. PMID- 8226925 TI - Inhibition of epidermal growth factor receptor aggregation by an antibody directed against the epidermal growth factor receptor extracellular domain. AB - We have examined the perturbation of epidermal growth factor (EGF) receptor receptor interactions by a monoclonal antibody (13A9) that binds to the receptor extracellular domain. While 13A9 did not inhibit EGF binding, it inhibited energy transfer between fluorescent-labeled EGF molecules bound to receptors in membranes from human A431 cells by 70-100%. This antibody also inhibited EGF stimulated receptor dimerization in membranes as assessed by chemical cross linking and Fab fragments of the antibody strongly inhibited the EGF-stimulated dimerization of solubilized receptors when assessed by velocity sedimentation. However, under conditions where 13A9 inhibited receptor-receptor interactions within the plasma membranes, the antibody had no effect on EGF-stimulated receptor autophosphorylation or tyrosine kinase activity toward an exogenous substrate. Moreover, although the antibody significantly inhibited receptor dimerization in A431 cells, it had no effect on EGF-stimulated changes in cytosolic free [Ca2+] or 125I-EGF uptake in these cells, or on EGF-stimulated DNA synthesis in Swiss 3T3 cells. We conclude that the dimerization of the EGF receptors in a membrane environment is not required for full activation of tyrosine kinase activity and that inhibition of the dimerization of a large fraction of EGF receptors in cells does not necessarily inhibit several EGF mediated cellular responses. PMID- 8226926 TI - Growth of melanocytic cells is associated with down-regulation of protein kinase C alpha, delta, and epsilon isoforms. Possible role of diacylglycerol. AB - Protein kinase C (PKC) down-regulation has been shown to correlate with the growth of murine melanocytic cells in culture (Brooks, G., Wilson, R. E., Dooley, T. P., Goss, M. W., and Hart, I. R. (1991) Cancer Res. 51, 3281-3288). We now show that PKC alpha, delta, epsilon, and zeta isoforms are present at the protein level in quiescent, non-transformed Mel-ab melanocytes, maintained in the absence of phorbol ester. Proliferation of Mel-ab cells, achieved by incubation in the continual presence of phorbol 12,13-dibutyrate, was associated with a down regulation of the PKC alpha, delta, and epsilon isozymes. Examination of two transformed syngeneic lines (the B16 murine melanoma and the long terminal repeat Ras.2 line), that grew in the absence of exogenous phorbol esters, showed that PKC alpha protein levels were either partially down-regulated or unaffected, the PKC delta and epsilon isoforms were down-regulated completely, and the levels of PKC zeta protein remained unaltered relative to quiescent Mel-ab cells. Basal levels of total diacylglycerol were elevated 5-fold in B16 melanoma cells compared with levels found in quiescent or proliferating Mel-ab melanocytes and appear to arise largely from the breakdown of phosphatidylinositol phospholipids accompanied by a significant rise in phospholipase C activity. Hourly treatments of quiescent Mel-ab melanocytes with the synthetic diacylglycerol analogue, 1,2 dioctanoyl-sn-glycerol, for 24 h, resulted in an induction of DNA synthesis which was associated with a significant down-regulation of PKC levels mediated largely via post-translational rather than transcriptional mechanisms. These results show for the first time that specific isoforms of PKC are down-regulated at the protein level during proliferation of murine melanocytic cells and suggest that the constitutive down-regulation of PKC in transformed melanoma cells may arise as a consequence of elevated endogenous phosphatidylinositol-derived diacylglycerol levels. PMID- 8226927 TI - Inactivation of rabbit muscle glycogen synthase by glycogen synthase kinase-3. Dominant role of the phosphorylation of Ser-640 (site-3a). AB - Rabbit skeletal muscle glycogen synthase, a rate-limiting enzyme for glycogen biosynthesis, is regulated by multisite phosphorylation. The protein kinase glycogen synthase kinase 3 (GSK-3) phosphorylates 4 Ser residues (Ser-640, Ser 644, Ser-648, and Ser-652; also known as sites 3a, 3b, 3c, and 4, respectively) at the COOH terminus of the subunit. Phosphorylation of these sites by GSK-3 is sequential, from COOH- to NH2-terminal, and is wholly dependent on prior phosphorylation by casein kinase II at Ser-656 (site 5). Expression in Escherichia coli was used to generate mutant forms of glycogen synthase, S640A, S644A, and S648A, in which site 3a, site 3b, or site 3c was changed to Ala, respectively. The purified enzymes had -/+ glucose-6-P activity ratios in the range of 0.8-0.9. Phosphorylation by casein kinase II and GSK-3 gave results consistent with the model of obligate sequential action of GSK-3. Phosphorylation at site 5, sites 4 + 5, or sites 3c + 4 + 5 had no measurable effect on activity. When sites 3b + 3c + 4 + 5 were phosphorylated, modest inactivation resulted. Additional phosphorylation at site 3a, however, was potently inactivating, reducing the -/+ glucose-6-P activity ratio to 0.1 and increasing the glucose-6-P concentration needed for half-maximal activation by an order of magnitude. Introduction of each additional phosphate, in the order site 4, 3c, 3b, and 3a, caused an incremental reduction in the mobility of the subunit when analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The results of this study demonstrate that GSK-3 phosphorylation of site 3a (Ser 640), and to a lesser extent, site 3b, correlates with inactivation of glycogen synthase by GSK-3. Evidence is also presented for an allosteric mechanism of inactivation whereby modification of one subunit influences the activity state of adjacent subunits. PMID- 8226928 TI - The pyruvate kinase gene as a model for studies of glucose-dependent regulation of gene expression in the endocrine pancreatic beta-cell type. AB - The insulinoma beta-cell line INS-1 expresses the L-type pyruvate kinase gene at high level and responds to a rise in extracellular glucose by strong induction of gene expression. Following the addition of glucose to the culture medium in the 3.5-33 mM concentration range, the cellular level of L-type pyruvate kinase mRNA increases within 2 h and reaches a maximum 15-fold above basal in 8-12 h. By run on nuclear assay, the relative transcription rate of the pyruvate kinase gene is shown to increase 4-fold at maximal stimulation, suggesting that both transcriptional and post-transcriptional effects contribute to mRNA accumulation. The glucose effect is totally suppressed by the hexokinase inhibitor mannoheptulose, indicating a requirement for glucose phosphorylation. The mRNA induction is not inhibited in glutamine-free culture medium or by azaserine, suggesting that the hexosamine biosynthetic pathway is not involved. Moreover, metabolism along the glycolytic pathway does not appear to be an absolute requisite, since 2-deoxyglucose partly mimics the inductive effect of glucose. The glucose effect on the pyruvate kinase gene is reversibly antagonized by agents increasing intracellular cAMP. In addition, the effect is highly specific to the pyruvate kinase gene. Neither proinsulin I mRNA nor glucokinase mRNA are increased in glucose-stimulated INS-1 cells. Short term transfection with CAT plasmids driven by the pyruvate kinase L promoter reveals specific glucose inducible reporter activity with the 183-base pair promoter region upstream of the cap site. Within this region, the previously described L4 cis-acting element is crucial for glucose responsiveness, as demonstrated by the fact that a plasmid with a mutation in this element does not elicit glucose-inducible CAT activity. Induction of L-type pyruvate kinase mRNA occurs in the islets of rats subjected to fasting and carbohydrate refeeding. In conclusion, the L-type pyruvate kinase gene provides an interesting model of glucose-regulated gene in the endocrine beta-cell type. PMID- 8226929 TI - Differential expression of transporters for norepinephrine and glutamate in wild type, variant, and WNT1-expressing PC12 cells. AB - Wild type PC12 pheochromocytoma cells express a Na(+)-dependent norepinephrine transporter that operates in the uptake of catecholamines, including dopamine. This transporter is not expressed in two spontaneously occurring flat cell variants of PC12 or in two other flat cell variants whose phenotype was induced by expression of the Wnt-1 oncogene. However, each of the flat cell variants, including those that express Wnt-1, exhibit a Na(+)-dependent, Cl(-)-independent glutamate/aspartate transporter activity that is not present in wild type PC12 cells. The flat cell variants took up glycine by a Na(+)-dependent process as well as did wild type cells. All of the flat cell variants have decreased levels of norepinephrine transporter mRNA but normal levels of glycine transporter mRNA. Glutamate/aspartate transporter mRNA was detected only in the variants that exhibited glutamate/aspartate transporter activity, and the nucleotide sequence of a partial glutamate/aspartate transporter cDNA from these cells demonstrated that it was the glial form of the transporter that was expressed. These variants were more sensitive than was wild type PC12 to alanosine, a toxic aspartate analog that enters cells by a transporter-mediated system such as the glutamate/aspartate transporter; however, these variants were as sensitive as wild type cells to another toxic aspartate analog, N-(phosphonacetyl)-L-aspartic acid, which is believed to enter cells by endocytosis. We suggest that the Wnt-1 gene product, or a homolog, may be involved in glial differentiation and that the mechanisms that alter the expression of the norepinephrine and glutamate/aspartate transporters in wild type and variant PC12 cells may also operate to regulate neurotransmitter transporter expression in vivo. PMID- 8226930 TI - Minimal sequence in heparin/heparan sulfate required for binding of basic fibroblast growth factor. AB - Experiments based on interaction in free solution between basic fibroblast growth factor (FGF-2) and saccharides related to heparin/heparan sulfate showed that the growth factor binds to heparin and to selectively glucosaminyl 6-O-desulfated heparin but poorly to iduronosyl 2-O-desulfated heparin. 2-O-sulfate groups thus are essential to the interaction, whereas 6-O-sulfates are not required nor do they interfere with FGF-2 binding. Comparison of various bound/nonbound oligosaccharides implicated a minimal pentasaccharide sequence for FGF-2 binding, with the structure: -hexuronic acid-glucosamine N-sulfate-hexuronic acid glucosamine N-sulfate-iduronic acid 2-O-sulfate- (reducing terminus to the right). Such (overlapping) sequences are abundant in heparin, albeit heavily obscured by irrelevant O-sulfate groups, and occur also in heparan sulfate, with or without additional O-sulfates. PMID- 8226931 TI - Cooperation of GATA-1 and Sp1 can result in synergistic transcriptional activation or interference. AB - GATA-1 is a lineage-restricted transcription factor. Virtually all erythroid expressed genes contain GATA recognition sites in their regulatory elements. Cotransfection/transactivation assays have revealed that, although GATA-1 as the only cell-restricted transcription factor is sufficient to activate some of the erythroid-specific promoters, not all such promoters are responsive, suggesting a requirement for cooperation with other factors. To study the interaction of GATA 1 with other transactivators, we analyzed sequence motifs of the human gamma globin promoter as response system by in vitro transcription and by transfections into erythroid K562 cells or into heterologous Drosophila SL2 cells. GATA-1 alone did not activate the promoter. However, GATA-1 exerted an effect in concert with the ubiquitous transactivator Sp1. Depending on the factor concentrations and the sequence context of the cognate binding sites, this interaction could result in synergistic transcriptional activation or in interference. GATA-1 and Sp1 did not cooperate in DNA binding when tested in vitro. This suggests that the functional cooperation is mediated by protein interactions with additional factor(s) which transmit the activator signal. The Sp1-binding CCACCC motif was found to be critical for high activity of the gamma-globin promoter. This site overlaps with a recognition sequence for members of the NFI/CTF family. NFI did not transactivate, but it interfered with Sp1-mediated stimulation and hence with Sp1/GATA-1 cooperation. These data, together with phylogenetic evidence, suggest that the CCACCC region is likely to represent a regulatory switch element. PMID- 8226932 TI - Decreased activities of phosphatidate phosphohydrolase and phospholipase D in ras and tyrosine kinase (fps) transformed fibroblasts. AB - The activity of N-ethylmaleimide-insensitive phosphatidate phosphohydrolase (PAP 2) was characterized in control, ras-transformed, and tyrosine kinase-(fps) transformed rat fibroblasts. PAP-2 was assayed in two different ways: 1) within its natural membrane using liposomes of phosphatidate and 2) in the presence of sufficient Triton X-100 to solubilize PAP-2, and to form mixed micelles with the phosphatidate. Harvesting the fibroblasts in medium containing orthovanadate and Zn2+ gave up to 3-fold higher PAP-2 activities when measured in the absence, but not in the presence, of Triton X-100. PAP-2-specific activities from both assays increased in the control fibroblasts as the cells reached confluence. Both specific activities were lower in the oncogenically transformed fibroblasts than in controls at all cell densities tested. The specific activities of PAP-2 did not increase with time in culture in transformed cells which continued to divide. The relative increase in activity of phospholipase D after stimulation with serum or phorbol myristate acetate was lower in the transformed fibroblasts compared to control cells. This indicates a coordinated decrease in the phospholipase D/phosphatidate phosphohydrolase pathway at the level of both enzymes in ras and fps transformed fibroblasts. The ratio of the production of diacylglycerol relative to phosphatidate, after stimulation with serum, or phorbol ester, was lower in both transformed fibroblasts relative to the controls. This is compatible with the decreased specific activity of PAP-2 and indicates functional significance for the differences in PAP-2 activity in regulating the balance between the two mitogenic lipids, phosphatidate and diacylglycerol. Control of PAP-2 activity could be an important factor in regulating appropriate signals for cell division. PMID- 8226933 TI - Properties of MEKs, the kinases that phosphorylate and activate the extracellular signal-regulated kinases. AB - Activation of extracellular signal-regulated kinase (ERK) or mitogen-activated protein kinase by MEK (mitogen-activated protein kinase or extracellular signal regulated kinase kinase) is an essential event in the mitogenic growth factor signal transduction. We now demonstrate that three recombinant MEKs (MEK1, MEK2, MEK3) show remarkably different activity toward recombinant ERK1 and ERK2. MEK2 is the most active ERK activator. The recombinant MEK1 has an activity approximately seven times lower than that of MEK2. MEK3, which is identical to MEK1 except for missing an internal 26-amino acid residue and probably represents an alternative splicing product of MEK1, shows neither autophosphorylation nor ERK-activating activity. Recombinant MEK1 and MEK2 can be activated by epidermal growth factor-stimulated SWISS3T3 cell lysate. MEK1 and MEK2 can also be activated by autophosphorylation. Autophosphorylation of MEKs correlates with their ability to phosphorylate and activate ERKs. Phosphorylation of MEK is also stimulated by ERK. Phosphoamino acid analysis showed that ERK1 preferentially phosphorylated threonine residue of MEKs. MEKs complex with ERKs in vitro. Interestingly, MEK3 also forms a complex with ERK1, although it is totally inactive as an ERK activator. PMID- 8226934 TI - Localization of the binding region of high mobility group protein 2 to cisplatin damaged DNA. AB - Cisplatin (CDDP) is an effective cancer chemotherapeutic drug used in the treatment of several human malignancies. The effectiveness of cisplatin therapy is limited by intrinsic resistance of tumors to this drug as well as the development of secondary tumors, which are also drug resistant. A potential mechanism influencing the sensitivity of cells to CDDP may result from the interaction of specific proteins with CDDP-damaged DNA (CDDP-DNA). In an earlier report, we demonstrated that high mobility group (HMG) proteins 1 and 2 bind with high affinity to CDDP-DNA. In the present study partial proteolytic digestion was used to localize the binding region of HMG2. A proteolytic fragment of approximately 20 kDa, containing the amino-terminal region of the protein, maintains the ability to bind with high affinity to CDDP-DNA, while an amino terminal fragment of 14 kDa binds with slightly reduced affinity. In contrast, a peptide fragment lacking 51 NH2-terminal amino acids from HMG2 has greatly reduced affinity for damaged DNA. Recombinant peptide fragments containing HMG box 1 or HMG box 2 bind weakly to damaged DNA, while a recombinant fragment containing HMG boxes 1 and 2 binds with high affinity. Hence, our results indicate that the amino-terminal region of HMG2 contains the damaged DNA binding recognition site and that both HMG boxes 1 and 2, present in the parental molecule, are required for high affinity binding of this protein to CDDP-DNA. PMID- 8226935 TI - Fibronectin receptors from Streptococcus dysgalactiae and Staphylococcus aureus. Involvement of conserved residues in ligand binding. AB - The nucleotide sequence of two genes encoding fibronectin (Fn) receptors FnBA and FnBB of Streptococcus dysgalactiae S2 revealed the presence of repeated motifs (called RA1-A3 and RB1-B3, respectively) which encode Fn binding activity (Lindgren, P.-E., McGavin, M. J., Signas, C., Guss, B., Gurusiddappa, S., Hook, M., and Lindberg, M. (1993) Eur. J. Biochem. 214, 819-827). Synthetic peptides of 32-37 amino acids, corresponding to individual repeated motifs, were assayed for the ability to inhibit Fn binding to cells of S. dysgalactiae. Within the RA motifs, peptide A2 was 10-fold more active than either A1 or A3, while in the RB motifs, only B3 was active. The same level of activity is observed when these synthetic peptides were assayed for inhibition of Fn binding to cells of Staphylococcus aureus. Likewise, synthetic peptides corresponding to the RD1-D3 motifs, which comprise a ligand binding domain in a Fn receptor from S. aureus, inhibit binding of Fn to both S. aureus and S. dysgalactiae. Assays of chemically modified peptides and peptide fragments derived from chemical or proteolytic cleavage suggest that a conserved core sequence, defined as ED(T/S) (X9,10)GG(X3,4)(I/V)DF, within a 30-amino acid-long segment is present in the active RA and RD motifs. Analyses of the importance of individual residues of this core sequence indicate that the ED(T/S) motif is nonessential, whereas the GG and the (I/V)DF together with additional acidic residues in the C-terminal half of the peptide are required for activity. PMID- 8226936 TI - Mechanisms of ferulic acid conversions to vanillic acid and guaiacol by Rhodotorula rubra. AB - Resting cells of Rhodotorula rubra converted transferulic acid (1) to vanillic acid (2), then to guaiacol (3) and protocatechuic acid (4), under aerobic conditions. In an argon atmosphere, R. rubra transformed ferulic acid to vanillic acid and 4-hydroxy-3-methoxystyrene (5). Metabolites were isolated by solid-phase extraction and characterized by mass spectrometry, 1H, and 13C-nuclear magnetic resonance spectroscopy (NMR). The biotransformation of ferulic acid to vanillic acid by R. rubra cell-free extracts required CoA, ATP, and NAD+. Mass spectrometry and 13C-NMR were used to demonstrate the incorporation of oxygen from H2(18)O during the conversion of ferulic acid to vanillic acid. The results suggest a parallel between this bioconversion reaction and the beta-oxidation of fatty acids. Proton-carbon correlation NMR spectroscopy was used to demonstrate the specific incorporation of deuterium from D2O into guaiacol obtained from vanillic acid. The incorporation of deuterium implicates the involvement of a quinoid vanillic acid tautomer as an intermediate in the decarboxylation reaction. PMID- 8226937 TI - Consequences of weak interaction of rho GDI with the GTP-bound forms of rho p21 and rac p21. AB - rho GDI is an inhibitory GDP/GTP exchange protein for the rho family. Recently, rho GDI has been reported to interact with the GTP-bound form of G25K and rac1 p21 and to inhibit their basal and GTPase-activating protein (GAP)-stimulated GTPase activity. Here, we examined whether rho GDI interacts with the GTP-bound form of rho p21 and rac p21 and inhibits their basal and rho GAP-stimulated GTPase activity, rho GDI interacted with both the GDP- and GTP-bound forms of rhoA p21 and rac1 p21 as estimated by measuring its ability to form a complex with both forms and to inhibit the membrane-binding activity of both forms. The efficiency of rho GDI for interaction with the GTP-bound form was, however, about 10% that for interaction with the GDP-bound form. Moreover, rho GDI inhibited both the basal and rho GAP-stimulated GTPase activity of rhoA p21 and rac1 p21 in a dose-dependent manner. The doses of rho GDI necessary for this action were, however, about 10-fold higher than those necessary for the action to inhibit their GDP/GTP exchange reaction. These results indicate that rho GDI interacts with the GTP-bound form of its substrate small G proteins, as well as with the GDP-bound form, but much less efficiently than with the GDP-bound form. PMID- 8226938 TI - Alternative splicing in a novel tyrosine phosphatase gene (DPTP4E) of Drosophila melanogaster generates two large receptor-like proteins which differ in their carboxyl termini. AB - A novel Drosophila receptor-like protein tyrosine phosphatase gene, DPTP4E, was isolated and characterized. DPTP4E, located at cytological position 4E1-2, is comprised of 10 exons; its RNA products are widely expressed during embryonic development, including the developing central nervous system. DPTP4E produces three major developmentally regulated transcripts of 6.5, 7.0, and 7.5 kilobases. The two major embryonic transcripts arise as the result of the alternative splicing of exon IX; as a consequence, two proteins (200 and 183 kDa) are produced which differ in their carboxyl-terminal sequences. The deduced extracellular domain, which lies between two putative hydrophobic transmembrane segments, contains 11 fibronectin type III-like repeats and 25 putative N glycosylation sites. A single conserved protein tyrosine phosphatase (PTPase) catalytic domain, which shows a high level of amino acid identity to the Drosophila PTPase DPTP10D and human HPTP beta, is found in the predicted intracellular domain; this PTPase domain, when expressed as a fusion protein in Escherichia coli, exhibits PTPase activity. The possible implications of these findings are discussed. PMID- 8226939 TI - Phosphorylation/dephosphorylation of the receiver module at the conserved aspartate residue controls transphosphorylation activity of histidine kinase in sensor protein ArcB of Escherichia coli. AB - A membrane sensor protein, ArcB, recognizes anaerobic environments and signals the information to the cognate regulator, ArcA. This in vitro study presents the process and control of the signal-transduction phosphorylation. In the presence of ATP, the ArcB transmitter module undergoes autophosphorylation and then transfers the phosphoryl group to its own receiver module as well as to the ArcA receiver module. Results suggest that the phosphoryl group of the ArcB receiver module is released by an intrinsic phosphatase activity. D-Lactate inhibits the phosphatase activity that removes phosphoaspartate groups from the receiver module in ArcB, and the associated increase in phosphorylation of this module leads to an activation of transphosphorylation of subsequently added phosphohistidine groups on ArcB to the receiver module of ArcA. A similar effect was also observed in the presence of pyruvate, acetate, or NADH. Conversely, the non-phosphorylated ArcB receiver module completely inhibits the intermolecular transphosphorylation. Thus, the phosphorylation state of the ArcB receiver module controls signal transduction from ArcB to ArcA. Since the intrinsic phosphatase activity is inhibited by cellular metabolites that increasingly accumulate by anaerobiosis, the enzyme portion of ArcB may be involved in sensing anaerobic environments through cellular metabolites in vivo. PMID- 8226940 TI - Site-directed mutagenesis of glutathione S-transferase YaYa. Mapping the glutathione-binding site. AB - Previous studies from our laboratory have shown that aspartic acid 101 plays an important role in glutathione interaction to rat glutathione S-transferase YaYa, while tyrosine 9 is directly involved in catalysis. Based on the available structural information, site-directed mutagenesis was conducted to examine the function of arginine, lysine, glutamine, and proline residues surrounding the GSH binding pocket. Arginine mutants R13K, R15K, R20K, and R20I retained partial enzymatic activities, while R13I and R15I lost most of their activities. Kinetic studies showed a marked increase in Km toward GSH for R15I suggesting that arginine 15 contributes significantly to the binding of GSH in the active site of glutathione S-transferase YaYa. A drastic decrease in enzymatic activities for R13I suggested the importance of the charged group of arginine 13 either in maintaining the structural integrity of the enzyme or in serving a vital role in enzymatic function. Replacement of glutamine 54 and 67 with glutamic acid or asparagine resulted in decreased enzymatic activities. Moreover, an 11-, 17-, and 9-fold increase in Km values toward GSH for mutant Q54E, Q54N, and Q67N was observed, respectively. These results suggested that glutamine 54 and 67 also contributed significantly to the binding of GSH. Proline at position 56 appears to be important for maintaining the structural integrity of the enzyme since mutants P56A and P56F were much less active and extremely less stable than that of the wild type enzyme. Both lysine mutants, K45R and K45I, exhibited substantially higher catalytic efficiencies toward both 1-chloro-2,4 dinitrobenzene and GSH than the wild type enzyme. Our data clearly show that lysine 45 is not an essential residue for catalysis nor for GSH binding in glutathione S-transferase YaYa. PMID- 8226941 TI - Addition of an endoplasmic reticulum retrieval sequence to ricin A chain significantly increases its cytotoxicity to mammalian cells. AB - An Escherichia coli expression system was used to produce recombinant ricin A chain (RTA) and RTA modified either by the addition of a carboxyl-terminal endoplasmic reticulum retrieval sequence Lys-Asp-Glu-Leu (RTAKDEL) or a nonfunctional analogue Lys-Asp-Glu-Ala (RTAKDEA). These RTA molecules can enter mammalian cells by fluid phase endocytosis. RTAKDEL was significantly more cytotoxic than either RTA or RTAKDEA to both Vero cells and HeLa cells (250- and 10-fold, respectively), despite the fact that all these RTA molecules had comparable enzymatic activities. This difference did not result from KDEL mediated binding of RTAKDEL to the cell surface. Enhanced cytotoxicity could be correlated with an increased level of ribosome inactivation, measured as the RTA catalyzed depurination of 28 S ribosomal RNA. These results indicate that the added KDEL sequence facilitated RTA entry into the cytosol. We propose that interaction with the intracellular KDEL receptor promotes retrograde transport of the toxin to the endoplasmic reticulum, where translocation of RTA into the cytosol occurs. PMID- 8226942 TI - Activation of a recombinase-deficient mutant recA protein with alternate nucleoside triphosphate cofactors. AB - We recently described two mutant recA proteins, (G160N)recA and (H163A)recA, which have full single-stranded DNA-dependent ATP hydrolysis activity but which are unable to promote the ATP-dependent strand exchange reaction under standard reaction conditions (pH 7.5). These mutant proteins, however, are able to promote strand exchange at pH 6.0 to 6.8. Here we show that this activation correlates with a pH-dependent decrease in the S0.5 value for ATP, with the (H163A)recA protein becoming active in strand exchange at pH values where the S0.5(ATP) decreases below 100 microM. We also show that the (H163A)recA protein is active in strand exchange over the range of pH 6.0-8.2 if dATP (or ddATP) is used in place of ATP as a cofactor; dATP is hydrolyzed by (H163A)recA protein at the same rate as ATP but has an S0.5 value lower than 100 microM across this pH range. These results are discussed with regard to the general significance of the S0.5 value in determining whether a nucleoside triphosphate will be able to stabilize the recA-single-stranded DNA filament in the strand exchange active conformational state. PMID- 8226943 TI - Amplification of bacteriophage Mu DNA by rolling circle DNA replication in vitro. AB - When mini-Mu DNA was allowed to transpose and replicate in vitro over a prolonged period, the products consisted not only of simple inserts and cointegrates but also high molecular weight DNA many times the unit length of mini-Mu. This high molecular weight product contained predominantly full-length mini-Mu DNA and relatively little non-Mu DNA (the vector harboring the mini-Mu element and the target for transposition in the reaction system). It arose from rolling circle DNA replication of templates created by intramolecular strand transfer, which is catalyzed by Mu transposition proteins. A donor substrate, which is a supercoiled plasmid bearing a mini-Mu element, gave rise to large amounts of the high molecular weight product provided that the vector segment outside the mini-Mu element was 2 kilobase pairs or more. When a donor substrate had a vector segment of only 600 base pairs, the mini-Mu element first had to transpose to a larger circular target before giving rise to the high molecular weight product. These results suggest a mechanism by which Mu DNA can be amplified for lytic development without transposing multiple times. By establishing a circular template, multiple copies of Mu can be processively generated from a single initiation event. PMID- 8226944 TI - Cytosine deaminase. The roles of divalent metal ions in catalysis. AB - Cytosine deaminase (CDase, EC 3.5.4.1) isolated from Escherichia coli contains a catalytically essential divalent metal ion. Fe2+ was efficiently removed from the enzyme with o-phenanthroline to yield an apoenzyme with less than 5% of the catalytic activity of native enzyme. The time courses for inactivation and for removal of Fe2+ from the enzyme by o-phenanthroline were similar. Apoenzyme reconstituted with Fe2+, Mn2+, Co2+, or Zn2+ (M2+CDase) had kcat values of 185, 88, 50, and 32 s-1, respectively. The Km values of these M2+CDases for cytosine were similar (0.22-0.39 mM). Cytosine potently inhibited reconstitution of the apoenzyme with Fe2+. Fe2+CDase was rapidly inactivated by 1 mM H2O2 (t1/2 < 1 s), whereas Mn2+CDase, Co2+CDase, and Zn2+CDase were not inactivated by H2O2. CDase was also inhibited by excess divalent cations. Cu2+ and Zn2+ reversibly inhibited Fe2+CDase activity with inhibition constants of 1.8 and 5.8 microM, respectively. Cu2+ dissociated slowly from the secondary binding on CDase with a rate constant of 2 x 10(-3) s-1. PMID- 8226945 TI - 1H NMR study of the dynamics of the pH modulation of axial coordination in Aplysia limacina (Val(E7)) and sperm whale double mutant His(E7)-->Val,Thr(E10)- >Arg metmyoglobin. AB - The molecular and electronic structure, thermodynamics, dynamics, and mechanism of interconversion of the pH-modulated reversible equilibria of Aplysia limacina metmyoglobin, (metMb), have been investigated by 1H NMR spectroscopy. The four identified species which interconvert slowly on the NMR time scale (lifetime > 1 ms) are metMbOH (B) at alkaline pH, five coordinate metMb (N) at acidic to neutral pH, an acidic form, A, near pH approximately 4 and an extremely low pH form, D, attributed to an equilibrium unfolded species. The presence of strong distal hydrogen bonding by Arg (E10) to bound hydroxide is detected via a significant solvent isotope effect on the metMbOH (B) hyperfine shifts. Integration of the peak intensities yields pK values of 7.7 and approximately 4 for the B<-->N and N<-->A equilibria, respectively. Saturation transfer via chemical exchange is observed for B<-->N and N<-->A, where the rates for forming metMbOH (B) and the acidic form A from N are base- and acid-catalyzed, respectively, while the reverse rates are first-order. The much slower interconversion rate for N<-->B in A. limacina metMb than His(E7) containing mammalian metMb is attributed to the fact that a ligand bond is broken rather than just proton transferred and that the equilibrium involves a major rearrangement of the orientation of Arg(E10). This conclusion is supported by 1H NMR data for the sperm whale double mutant His(E7)-->Val/Thr(E10)-->Arg metMb, which exhibits a pK approximately 8.7 for the equilibrium between five-coordinate metMb (N) and metMbOH (B) with an even slower interconversion rate than in A. limacina metMb. This double mutant metMbOH (B) exhibits hydrogen bonding by Arg (E10) with coordinated hydroxide similar to that in A. limacina metMbOH. The slow but acid-catalyzed rates of conversion of A. limacina metMb (N) to the acid species A with significantly weakened bonding of the heme iron to the axial His(F8) residue is consistent with protonation of an inaccessible residue and/or a structural change accompanying the protonation equilibrium. It is concluded that metMb will coordinate water strongly only when there is a distal hydrogen bond acceptor residue, while the hydroxide ion is coordinated strongly only if there is a distal hydrogen bond donor residue. PMID- 8226946 TI - Study of the conformation of DARPP-32, a dopamine- and cAMP-regulated phosphoprotein, by fluorescence spectroscopy. AB - DARPP-32 is a potent inhibitor of protein phosphatase 1 when it is phosphorylated on Thr34 by cAMP-dependent protein kinase. DARPP-32 is also phosphorylated on Ser45 and Ser102 by casein kinase II, resulting in a facilitation of phosphorylation by cAMP-dependent protein kinase. We have studied the conformation of recombinant rat DARPP-32 by steady-state and time-resolved fluorescence. The steady-state emission spectra and quenching of the intrinsic (Trp163) and extrinsic fluorescence (acrylodan or lucifer yellow linked to Cys72) were consistent with a complete exposure of these residues to the aqueous environment. The intrinsic fluorescence of DARPP-32 was resolved into three decay components with lifetimes of 1, 3.4, and 7 ns, with the intermediate lifetime component giving the major contribution. The ratio between the amplitudes associated with the short and long decay constants was decreased upon denaturation. The rotational behavior of DARPP-32 measured by anisotropy decay revealed that Trp163 is located in a highly flexible peptide chain, whereas Cys72 is embedded in a more rigid environment. Phosphorylation by cAMP-dependent protein kinase did not alter any of the fluorescence parameters, whereas only minor effects were associated with casein kinase II phosphorylation. These findings indicate that DARPP-32 contains at least two distinct domains and that phosphorylation has no dramatic effects on its conformation. PMID- 8226947 TI - Virus-mediated induction of interferon A gene requires cooperation between multiple binding factors in the interferon alpha promoter region. AB - Transcriptional activation of interferon A (IFNA) gene in virus-infected cells is controlled by a 35-nucleotide inducible element that is cell type specific. Within this region, two elements, alpha F1 and IRF-1 binding sites, were shown by mutation analysis to play a crucial role in the expression of inducible element. In this study, we have analyzed the binding of nuclear proteins to the alpha F1 sequence and have shown that the induction is associated with the formation of a novel complex alpha F1/B, which contains at least two DNA binding proteins of 68 and 96 kDa. In contrast, no binding of the purified interferon regulatory factor 1 (IRF-1) either to the alpha F1 or IRF-1 binding sites could be detected in vitro. However, the oligonucleotides corresponding to alpha F1 or IRF-1 binding sites competed efficiently for the induction of IFNA4 promoter region in a transient transfection assay. We suggest that the induction of IFNA promoter region requires cooperation between alpha F1 binding proteins and IRF-1. Interestingly, our data also show that the inability of IFNA6 promoter to be expressed in infected L-cells may be a result of a viral-induced repressor, which could act by binding and inactivating alpha F1 or by competing for the IRF-1 binding site. These results suggest that cell-specific expression of IFNA genes results from core-cruitment of trans-acting factors that bind to alpha F1 and the IRF-1 binding site with the cell-specific virus-induced activator or repressor. PMID- 8226948 TI - Factor IX Fukuoka. Substitution of ASN92 by His in the second epidermal growth factor-like domain results in defective interaction with factors VIIa/X. AB - Hemophilia B Fukuoka, a moderately severe bleeding disorder, is a naturally occurring mutant of factor IX. Plasma from our patient had 3% clotting activity even though 64% of factor IX antigen was present. The purified mutant protein was cleaved normally by factor Xla, factor VIIa-tissue factor complex, or RVV-X (factor X-activating enzyme from Russell's viper venom), yielding a two-chain factor IXa. Amino acid composition and sequence analyses of one of the lysyl endopeptidase peptides derived from factor IX Fukuoka revealed that Asn92 in the second epidermal growth factor (EGF)-like domain had been replaced by His. The active site of the factor IXa Fukuoka was normally competent for the incorporation of p-aminobenzamidine and for the hydrolysis of a synthetic substrate, N alpha-benzyloxycarbonyl-L-arginine p-nitrobenzyl ester. Factor Xa formation by factor IXa Fukuoka was only 8% of the normal factor IXa, even in the presence of polylysine, and only 0.2% of the normal in the system containing phospholipids, Ca2+, and factor VIIIa, thereby indicating a functional defect in interaction of the mutant with factors VIIIa/X. Furthermore, catalytic efficiency (kcat/Km) of factor IXa Fukuoka toward factor X in the presence of Ca2+, phospholipids, and factor VIIIa was only 2.3% of the normal factor IXa. These results suggest that an Asn-to-His substitution at position 92 in the second EGF like domain of factor IX Fukuoka would have an untoward effect on the specific conformational state of factor IX for binding with factors VIIIa/X. PMID- 8226949 TI - Purification and characterization of a general transcription factor, aTFB, from the archaeon Methanococcus thermolithotrophicus. AB - We have recently shown that cell-free transcription of homologous templates from the archaeon Methanococcus thermolithotrophicus requires an archaeal transcription factor (aTFA) that separated from the RNA polymerase during phosphocellulose chromatography. We report here the identification and extensive purification of a second activity, aTFB, required for in vitro transcription. This activity copurified with RNA polymerase during initial chromatographic steps but was positively identified as a distinct transcription factor after Superdex 200 sizing chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the intensity of a M(r) = 28,000 polypeptide in silver-stained gels is correlated with transcription factor activity. The same polypeptide, when eluted from a denaturing polyacrylamide gel and subsequently renatured, showed the functional properties of the transcription factor. In conjunction with gel filtration and sedimentation studies, which indicated a molecular mass of 54,000 Da for the native protein, these results suggested that aTFB is a dimer with polypeptide chains of identical molecular mass. Functional studies with highly purified aTFB demonstrated that it is a general factor required for transcription of genes encoding tRNA and proteins. PMID- 8226950 TI - Cholesterol and cholesterol esters: host receptors for Pseudomonas aeruginosa adherence. AB - Bacterial adherence to tissues is a necessary step in infection and results from interactions between surface molecules on the bacteria (adhesins) and plasma membrane receptors on host cells. Using a clinical isolate of Pseudomonas aeruginosa, we have examined the receptors required for adherence to Chinese hamster ovary (CHO) cells. In mutant CHO cell lines and mucin-producing lung epithelial cell lines, adherence did not depend on glycoproteins, glycolipids, or proteoglycans. Treating cells with glutaraldehyde and proteases also did not affect adherence. However, treating cells with ethanol diminished adherence and resulted in removal of factors that would facilitate binding of bacteria when coated on plastic culture plates. These factors were purified by solvent extraction and silica gel chromatography and were identified as cholesterol and cholesterol esters by coelution with authentic standards, gas chromatography, 1H NMR, and mass spectrometry. Bacterial adherence was reduced to wild-type CHO cells treated with Lovastatin and to cholesterol-requiring insect cells grown in cholesterol-deficient medium, suggesting that the bacteria bound to plasma membrane cholesterol. Thus, this clinical isolate of Pseudomonas uses cholesterol and cholesterol esters for adherence. We propose that binding to cholesterol and cholesterol esters may affect the pathogenicity of similar Pseudomonas strains. PMID- 8226951 TI - Defective galactofuranose addition in lipophosphoglycan biosynthesis in a mutant of Leishmania donovani. AB - A mutant cell line of Leishmania donovani (R2D2), previously selected for resistance to the cytotoxic lectin ricin agglutinin, was found to be totally deficient in the synthesis and expression of lipophosphoglycan, a dominant surface virulence factor. The metabolic defect in R2D2 parasites responsible for its lipophosphoglycan (LPG-) phenotype was investigated in this study. Following metabolic labeling of R2D2 parasites with either [3H]galactose or [3H]mannose, the main glycosylphosphatidylinositide product that accumulated was Glc-PO4-Man Man-GlcN-lyso-1-O-alkylphosphatidylinositol (PI). The metabolic defect was further defined using a cell-free glycosylation system. When membrane preparations from wild-type cells were incubated with UDP-[3H]galactose and unlabeled GDP-mannose in the absence of exogenous acceptors, radiolabeled lipophosphoglycan was synthesized. The addition of exogenous Man-Man-GlcN-PI or Galf-Man-Man-GN-PI stimulated lipophosphoglycan synthesis in vitro. In contrast, when membrane preparations from R2D2 cells were incubated with exogenous Man-Man GlcN-PI as an acceptor or in the absence of exogenous acceptor, the truncated glycosylphosphatidylinositide Glc-PO4-Man-Man-GlcN-PI was the main radioactive product synthesized. However, when exogenous Galf-Man-Man-GN-PI was added to the R2D2 in vitro system, radioactive lipophosphoglycan was synthesized. Collectively, these results indicate that the mutant R2D2 cells are unable to complete the assembly of the glycan core of LPG because of a defect in the synthesis of the "activated" galactofuranosyl donor or the lack of a functional galactofuranosyltransferase. PMID- 8226952 TI - Differential recovery of Ca2+ binding activity in mutated EF-hands of cardiac troponin C. AB - Previous studies showed that conversion of the first Ca2+ ligand in Ca(2+) binding sites III and IV from Asp to Ala decreased the affinity of cardiac TnC (cTnC) for the thin filament. Here, the functional consequences of mutation of the second ligand in the Ca(2+)-binding sites of cTnC were determined. Equilibrium dialysis and Tyr fluorescence studies showed that conversion of the second Ca2+ ligand to Ala (Asp-67, site II; Asn-107, site III; and Asn-143, site IV) inactivated all three Ca(2+)-binding sites in the free protein. Ca2+ binding to the mutated site II was not recovered upon association with a troponin complex, and proteins with this mutation were unable to regulate Ca(2+)-dependent ATPase activity in TnC-extracted myofibrils. However, Ca2+ binding was recovered at the mutated sites III and IV under the same conditions. Sequential addition of active and inactive cTnC proteins in a myofibril ATPase assay suggested that that Mg2+ binding was not recovered and that the recovered Ca2+ affinity of the mutated sites III and IV was much lower than that of the wild type in that the Ca2+ concentrations required for apparent thin filament binding by proteins containing mutations at sites III and/or IV were significantly greater than that required for the wild-type protein. PMID- 8226953 TI - The high affinity state of inositol 1,4,5-trisphosphate receptor is a functional state. AB - Inositol 1,4,5-trisphosphate (InsP3) is a second messenger responsible for the rapid and discontinuous release of Ca2+ from intracellular stores. In this study, the effects of the sulfhydryl reagent thimerosal were investigated on Ca2+ mobilization and on InsP3 binding. Thimerosal was shown to release Ca2+, in a dose-dependent manner, with an EC50 of 135.8 +/- 5.2 microM, from bovine adrenal cortex microsomes. Thimerosal-induced Ca2+ release was not prevented by heparin (250 micrograms/ml), ruling out a participation of InsP3 receptor in that effect. The slow rate of thimerosal-induced Ca2+ release rather suggested an inhibition of microsomal Ca2+ ATPase. At submaximal concentration, thimerosal (100 microM) was also shown to potentiate the release of Ca2+ induced by InsP3. Dose-response experiments revealed that thimerosal enhanced the apparent affinity of InsP3 by a factor 2.21 +/- 0.28, without modifying the maximal amount of Ca2+ released by InsP3. Thimerosal also enhanced, in a dose-dependent manner, [3H]InsP3 binding to adrenal cortex microsomes (EC50 = 43.3 +/- 7.6 microM). A similar effect was also observed on [3H]InsP3 binding to solubilized receptors, suggesting a direct modification of the receptor protein by thimerosal. The effects of thimerosal on Ca2+ release and [3H]InsP3 binding were abolished in the presence of the reducing agent dithiothreitol (1 mM), suggesting a modification by thimerosal of specific thiol groups on these microsomal proteins. Scatchard analysis revealed that thimerosal (100 microM) increased InsP3 receptor affinity by 1.87 +/- 0.26-fold. Kinetic analysis indicated that this increased affinity was due to an enhancement of InsP3 association rate constant. The concomitant increases of binding affinity and Ca2+ releasing potency suggest that the high affinity state of InsP3 receptor is a functional state. PMID- 8226954 TI - Photoaffinity labeling of the 45-kDa and 55-kDa forms of phosphatidylinositol 4 kinase from the yeast Saccharomyces cerevisiae. AB - The membrane-associated 45- and 55-kDa forms of phosphatidylinositol (PI) 4 kinase (ATP:phosphatidylinositol 4-phosphotransferase, EC 2.7.1.67) from Saccharomyces cerevisiae are inhibited by ADP by a competitive mechanism with respect to ATP. We initiated studies toward defining the ATP and ADP sites on the PI 4-kinases using azidonucleotide photoaffinity labeling probes. The photoprobe 8-azido-ATP fulfilled the criteria of a specific photoaffinity label for the 45- and 55-kDa PI 4-kinases. 8-Azido-ATP was a substrate and a competitive inhibitor of the PI 4-kinases with Ki values similar to the Km for ATP. 8-Azido-ATP photoinactivated the enzymes and was photoincorporated into the enzymes in a dose dependent manner at concentrations similar to the Ki values for the photoprobe. ATP, the true substrate, provided specific protection against photoinactivation and photoincorporation of the PI 4-kinases with 8-azido-ATP, whereas GTP, a nonspecific nucleotide, provided no protection against photoinactivation and photoincorporation. Photoaffinity labeling of the PI 4-kinases with 8-azido-ATP was specifically prevented with ADP. The photoprobe 8-azido-ADP also fulfilled the criteria needed to validate its use as a specific photoprobe for the PI 4 kinases. Photoinactivation of the PI 4-kinases with 8-azido-ADP was prevented specifically with ATP. Taken together, these data supported the conclusion that the ATP and ADP sites on the membrane-associated 45- and 55-kDa PI 4-kinases from S. cerevisiae were the same. PMID- 8226955 TI - Structure-function analysis of the hormone binding domain of the human estrogen receptor by region-specific mutagenesis and phenotypic screening in yeast. AB - To investigate the structural requirements for recognition and response to ligands by the human estrogen receptor (hER), a series of point mutations were generated in the hormone binding domain (HBD) of the receptor using a limited formic acid treatment of its cDNA. Receptors having a reduced sensitivity to the estrogen, 17 beta-estradiol (E2), or to the antiestrogen, trans-hydroxytamoxifen, were selected from a library of intact hER cDNAs containing these mutant HBDs by expression and phenotypic screening in yeast (Saccharomyces cerevisiae). Several were sequenced, and the encoded receptors were characterized in both yeast and mammalian (Chinese hamster ovary) cells using hormone-binding and transactivation assays. In general, parallel phenotypes were observed in yeast and in Chinese hamster ovary cells following estrogen exposure. We report on 15 receptors having point mutations located at various positions throughout the HBD. Four categories of mutants were identified: 1) those showing no change from wild type in their response to E2; 2) those showing a greatly reduced transactivation response over the range of ligand concentrations tested; 3) those requiring much higher concentrations of E2 for maximal transactivation, indicating a reduced sensitivity to ligand; and 4) those showing reduced response to E2, but little change in response to trans-hydroxytamoxifen in yeast. Two mutations in the carboxyl terminus of the HBD eliminated hormone-dependent transactivation despite the continued ability to bind E2 with high affinity. Therefore, our results show a separation of the transactivation and hormone-binding functions of the hER, and indicate that the integrity of many regions throughout the large, approximately 250-amino acid HBD is important for these functions. Our studies also demonstrate the advantages of using regional mutagenesis combined with phenotypic screening in yeast to complement site-directed mutagenesis when investigating a large, functionally complex region. PMID- 8226956 TI - A gene encoding a chloroplast omega-3 fatty acid desaturase complements alterations in fatty acid desaturation and chloroplast copy number of the fad7 mutant of Arabidopsis thaliana. AB - Mutations at the fad7 locus of Arabidopsis thaliana (previously called fadD) cause decreased desaturation of dienoic fatty acids in chloroplast lipids in plants grown at elevated temperatures. This suggested that the fad7 locus encodes a chloroplast omega-3 desaturase that catalyzes the desaturation of lipid-linked 18:2 and 16:2 fatty acids. In order to clone the fad7 gene, it was first genetically mapped relative to the flanking restriction fragment length polymorphism markers 4547 and 2488A on chromosome 3, and yeast artificial chromosomes covering the locus were identified. A putative desaturase cDNA clone that was isolated by low stringency heterologous probing with a cDNA for an endoplasmic reticulum-localized omega-3 desaturase (fad3) hybridized to the yeast artificial chromosomes and could not be resolved from the locus by restriction fragment length polymorphism mapping. Expression of the cDNA in transgenic fad7 mutant plants resulted in restoration of wild type fatty acid composition and suppression of a previously observed effect of the fad7 mutation on chloroplast number, indicating genetic complementation. The structural gene contained seven introns within a coding sequence of 1338 base pairs, which encodes a 446-amino acid polypeptide of 51,172 daltons. The amino-terminal region of the fad7 gene product contained a consensus chloroplast transit peptide. Except for the amino terminal domain, the deduced amino acid sequence of the fad7 gene product had high homology to the fad3 gene product, indicating that fad7 encodes an omega-3 desaturase and that the two genes arose from a common ancestral gene. There was no apparent effect of growth temperature on the steady-state levels of fad7 mRNA in wild type plants. PMID- 8226957 TI - Phi 29 DNA polymerase active site. Residue ASP249 of conserved amino acid motif "Dx2SLYP" is critical for synthetic activities. AB - phi 29 DNA polymerase shares with other alpha-like DNA polymerases several regions of amino acid sequence similarity and sensitivity to inhibitors of eukaryotic DNA polymerase alpha. In this paper, site-directed mutants in the phi 29 DNA polymerase residues Asp249, Ser252, Leu253, and Pro255 of the conserved amino acid motif "Dx2SLYP" are described. Two mutants, D249E and S252R, were drastically affected in all the synthetic activities, whereas their 3' to 5' exonuclease activity and interaction with the TP primer was normal. Mutant D249E, slightly affected in template-primer binding, was completely inactive in all conditions tested, suggesting that Asp249 could be playing a direct role in catalysis. On the other hand, mutant S252R, strongly affected in template-primer binding, showed some DNA polymerization activity in the presence of Mn2+. Mutants S252G and P255S showed a reduced template-primer binding ability; these mutants, together with mutant L253V, showed metal ion-dependent phenotypes in their synthetic activities and altered sensitivities to the PPi analog phosphonoacetic acid. All these results support the hypothesis that the Dx2SLYP motif forms part of the polymerization active site of the phi 29 DNA polymerase, being the Asp249 residue critical both for protein-primed initiation and DNA polymerization. PMID- 8226958 TI - Developmental, nutritional, and hormonal regulation of tissue-specific expression of the genes encoding various acyl-CoA dehydrogenases and alpha-subunit of electron transfer flavoprotein in rat. AB - Short chain (SCAD), medium chain (MCAD), and long chain acyl-CoA dehydrogenases (LCAD) catalyze the first step of fatty acid oxidation, while isovaleryl-CoA dehydrogenase (IVD) is involved in leucine oxidation. They are homologous flavoproteins belonging to the acyl-CoA dehydrogenase (ACD) family. Electron transfer flavoprotein (ETF) serves as an obligatory electron acceptor for these reactions. We demonstrated that the expression of SCAD, MCAD, and LCAD and the alpha-subunit of ETF (alpha-ETF) showed a similar developmental pattern, while that of IVD was distinctly different from others. The ontogenic pattern of each enzyme in the liver differed distinctly from that in the heart. The degree of glucagon-enhanced ACD expression in vivo and in vitro in both the liver and heart was especially high in fasted rats. Dexamethasone induced all ACD mRNAs in the heart. In contrast, it strongly suppressed mRNAs of all ACDs and alpha-ETF mRNA in the liver, except IVD mRNA. Dexamethasone induced IVD mRNA in both the liver and heart. Starvation strongly stimulated expression of all five genes in various tissues, with the highest in the heart, except the IVD gene which was down regulated. The degree of induction by 3-day starvation differed in different age groups of rats. Feeding the rats a fat-free diet for 7 days caused a marked increase of IVD mRNA in the heart, whereas the high fat diet for the same period resulted in a severe decrease of the same degree, suggesting a protein-sparing mechanism. However, these manipulations of dietary fat content had little effect on the expression of other ACD genes. PMID- 8226959 TI - The cAMP response element in the rat thyrotropin receptor promoter. Regulation by each decanucleotide of a flanking tandem repeat uses different, additive, and novel mechanisms. AB - A decanucleotide tandem repeat (TR) sequence, between -162 and -140 base pairs (bp) of the minimal thyrotropin receptor promoter, decreases gene expression by repressing constitutive enhancer activity of its cAMP response element (CRE). Each decanucleotide acts additively. CRE-binding proteins and liver or thyroid nuclear extracts footprint a region including the CRE and the 3' decanucleotide, 148 to -124 bp; nuclear proteins interacting with the 3' decanucleotide protect a smaller region -148 to -135 bp. Separate groups of nuclear proteins interact with the CRE and the 3' decanucleotide; mutations of the CRE affect protein interactions with the 3' decanucleotide and the converse. Nuclear proteins bind to single- or double-stranded 3' decanucleotide DNA; those interacting with the CRE bind only double-stranded DNA. The repressor action of the 5' decanucleotide is associated with an interaction between the coding strand and a single-stranded binding protein in liver and thyroid nuclear extracts. The 5' decanucleotide is in a CT-rich region with S1 nuclease hypersensitivity, near perfect mirror images, and direct repeats. The data therefore indicate that each TR decanucleotide modulates CRE constitutive enhancer activity by different but additive mechanisms, competition versus interaction with a single-stranded binding protein, and each interacts with different nuclear proteins that are not thyroid-specific. The same region in the human thyrotropin receptor represses CRE constitutive enhancer activity by the same mechanisms, despite a nonidentical sequence and no overt TR. PMID- 8226960 TI - Cyclic AMP selectively increases renin mRNA stability in cultured juxtaglomerular granular cells. AB - This study was undertaken to examine the regulation of renin release and gene expression in primary cultures of juxtaglomerular granular (JGG) cells. JGG cells, isolated from mouse kidney, demonstrated high purity and showed regulated renin release in vitro. Changes in steady-state renin mRNA levels were assessed by quantitative polymerase chain reaction techniques, with polymerase chain reaction amplification efficiency monitored by co-amplification of experimental samples with a dilution series of cDNA for a mutant template. When the cells were incubated in the presence or absence of forskolin, isoproterenol, or 8-bromo-cAMP plus 3-isobutyl-1-methylxanthine for 24 h or cholera toxin for 12 h, renin mRNA levels were increased 3.9-, 4.4-, 5.1-, and 3.3-fold, respectively (all, p < 0.05). A significant increase in renin mRNA levels was observed 8 h after treatment with forskolin, but no change was detectable at 4 h. Cycloheximide did not prevent the increase in renin mRNA by isoproterenol. When RNA synthesis was inhibited by incubation with actinomycin D (5 micrograms/ml), renin mRNA levels declined with a half-life of 3.0 +/- 0.8 h. Treatment with forskolin increased renin mRNA half-life to 10.8 +/- 2.7 h (p < 0.025). The half-life of beta-actin, endothelin-1, or the facilitative glucose transporter-1 (GLUT-1) mRNA expressed in the same cells was not altered, although the steady-state levels of GLUT-1 mRNA increased 2.2-fold after treatment with forskolin. These data demonstrate that cAMP increases renin release and mRNA levels in JGG cells in vitro, that the stimulatory effect of cAMP on renin mRNA is delayed but does not require new protein synthesis, and that the increased renin mRNA levels induced by cAMP are due in part to a selective increase in renin mRNA stability. PMID- 8226961 TI - Different assembly species of IgM are directed to distinct degradation sites along the secretory pathway. AB - In 38C B lymphocytes the membrane form of IgM is displayed on the cell surface whereas the secretory form of IgM is degraded. In the EH cell line, a light chain deficient variant of 38C cells, the mu heavy chains are partially assembled with surrogate light chains characteristic of pre-B cells. In these cells neither the membrane (microns) nor the secretory (microsecond) forms of the mu heavy chain reach their final destination, and both are rapidly degraded. The degradation of mu chains in EH cells, like that of microsecond in 38C cells, is nonlysosomal and occurs prior to the trans-Golgi. However, while microsecond degradation in 38C cells is inhibited by brefeldin A, in EH cells microsecond and micron are retained and degraded by a brefeldin A-insensitive mechanism. These results indicate that degradation in EH cells occurs within the endoplasmic reticulum, whereas degradation in 38C cells requires exit from this compartment. Thus, mu heavy chains can be degraded in multiple sites along the secretory pathway. The location of the degradation process is determined by the developmentally regulated assembly species of the mu chains with either "classical" or surrogate light chains. PMID- 8226962 TI - Expression of active human DNA ligase I in Escherichia coli cells that harbor a full-length DNA ligase I cDNA construct. AB - A recombinant plasmid for expression of full-length human DNA ligase I (phLig-I) was constructed in a plasmid/phage chimeric vector, pTD-T7N, which was derived from pUC118 by oligonucleotide-directed mutagenesis. The insert contained a 2757 base pair coding sequence for a whole human DNA ligase I and an extra ACC codon adjacent to the ATG initiation codon. This ACC codon was required for achieving high levels of expression of full-length DNA ligase I in Escherichia coli strain BL21. The recombinant plasmid, which was designed to exploit the T7 late promoter and the ATG initiation codon for beta-galactosidase was transfected into E. coli BL21 cells that express T7 RNA polymerase. The recombinant clone produced relatively high levels of DNA ligase I with a molecular mass of 130 kDa, as estimated by SDS-polyacrylamide gel electrophoresis. The DNA ligase was purified to near-homogeneity by the two-step column chromatographic procedure from BLphLig I cells that had been induced with isopropyl beta-D-thiogalactoside. The specific activity, chromatographic behavior, kinetic properties, molecular mass, and antigenicity of the recombinant human DNA ligase I were indistinguishable from those of purified mammalian DNA ligase I. Metabolically labeling experiments with 32P(i) indicate that the recombinant DNA ligase I was present as an enzyme-AMP reaction intermediate, but not as a phosphoprotein, in the E. coli cells. PMID- 8226963 TI - Mutational studies of human DNA polymerase alpha. Identification of residues critical for deoxynucleotide binding and misinsertion fidelity of DNA synthesis. AB - Conserved site-directed mutations were introduced into the second most conserved amino acid region, region II, of the human DNA polymerase alpha catalytic subunit. These mutants were expressed in the baculovirus system and purified to near homogeneity. The mutants had polymerase activity ranging from 4 to 60% compared with the wild type polymerase alpha. Steady-state kinetic analysis of mutants G841A, D860A, D860S, D860N, Y865S, and Y865F demonstrated no significant difference in their Km values for primer-template compared with that of the wild type enzyme. In contrast, mutants D860A, Y865S, and Y865F showed a 5-10-fold increase in the Km for deoxynucleotide triphosphate (dNTP) compared with the wild type enzyme. DNA synthetic fidelity studies of these mutants showed that mutant Y865S but not Y865F had a greater than 10-fold higher misinsertion efficiency than the wild type enzyme in Mg(2+)-catalyzed reactions. However, with Mn2+ as the metal activator, Y865S and Y865F demonstrated a 2- and 9-fold higher misinsertion efficiency, respectively. These results indicate that Asp860 and Tyr865 in region II of human DNA polymerase alpha are involved in incoming dNTP substrate binding. Using three deoxynucleotide structural analogs as probes, we show that the nucleotide base is the structural requirement for dNTP binding with Tyr865. Furthermore, abolishing the hydrophobic phenyl ring side chain of Tyr865 by replacing tyrosine with serine rendered the enzyme resistant to aphidicolin. Results of these studies strongly suggest that the phenyl ring of Tyr865 directly interacts with the nucleotide base moiety of the dNTP and plays a critical role in the misinsertion fidelity of DNA synthesis. Although mutation of Gly841 to Ala did not affect the binding of primer-template, it had a significant decrease in kcat, an increase in Km for dNTP, a striking decrease of processivity, and also resistance to aphidicolin. Thus, mutation of this residue, Gly841, which is highly conserved among the alpha-like DNA polymerases, appears to affect both catalysis and substrate deoxynucleotide binding. This suggests that Gly841 is essential for the maintenance of the overall structure of the polymerase alpha catalytic site. PMID- 8226964 TI - Mutational studies of human DNA polymerase alpha. Serine 867 in the second most conserved region among alpha-like DNA polymerases is involved in primer binding and mispair primer extension. AB - The second most conserved region of alpha-like DNA polymerases, region II, spans a block of 40 amino acid residues centered at the core sequence -DFNSLYPSII-. In the previous paper, we described mutational studies of 3 amino acid residues in region II which includes 2 amino acid residues in the core sequence. We showed that residues Asp860 and Tyr865 in the core sequence are involved in substrate deoxynucleotide triphosphate (dNTP) binding. We further showed that the phenyl moiety of the Tyr865 side chain interacts with the incoming dNTP and is responsible for the misinsertion fidelity of the enzyme. In this report, we investigated the function of 2 serine residues, Ser863 and Ser867, in this core sequence. Mutation of these 2 Ser residues to either Ala or Thr yielded mutant enzymes with similar Km for dNTPs, kcat, processivity, and misinsertion fidelity of DNA synthesis as the wild type enzyme. However, mutation of Ser867 to Ala demonstrated a 30-fold increase in Km for primer-template and a 5-fold higher KD for binding primer-template. DNA footprinting experiments of primer with the dideoxynucleotide terminus indicated that the structural feature of the primer recognized by Ser867 is the 3'-OH terminus. Single-stranded DNA inhibition data suggest that removal of the hydroxyl side chain of Ser867 affects the polymerase's interaction with primer and not with template. Mutation of Ser867 to Ala also decreases the mutant enzyme's Km for dNTP to extend a mispaired primer and thus enhances its capacity to extend a mispaired primer terminus. These data support the conclusion that the hydroxyl side chain of Ser867 of human DNA polymerase alpha is involved in primer interaction during DNA synthesis and plays an essential role in mispair extension fidelity of DNA synthesis. PMID- 8226965 TI - The mechanism of inhibition of glycosylphosphatidylinositol anchor biosynthesis in Trypanosoma brucei by mannosamine. AB - The inhibition of glycosylphosphatidylinositol anchor biosynthesis by mannosamine has been described previously in the procyclic forms of Trypanosoma brucei and in mammalian cells (Lisanti, M. P., Field, M. C., Caras, I. W. J., Menon, A. K., and Rodriguez-Boulan, E. (1991) EMBO J. 10, 1969-1977). A recent report has suggested that mannosamine exerts these effects by becoming incorporated into glycosylphosphatidylinositol anchor intermediates (Pan, Y-T., Kamitani, T., Bhuvaneswaran, C., Hallaq, Y., Warren, C. D., Yeh, E. T. H., and Elbein, A. D. (1992) J. Biol. Chem. 267, 21250-21255). In this paper we have analyzed the effects of mannosamine on glycosylphosphatidylinositol anchor and variant surface glycoprotein biosynthesis in the blood-stream form of T. brucei. Trypanosomes were biosynthetically labeled with [3H]mannosamine, and [3H]glucosamine in the presence of mannosamine, and the structures of the labeled glycolipids which accumulated were determined. The main glycolipid metabolite of mannosamine was shown to be ManN-Man-GlcN-PI. A trypanosome cell-free system preloaded with this compound was significantly impaired in its ability to synthesize glycosylphosphatidylinositol anchor intermediates beyond Man alpha 1-6Man alpha 1 4GlcN alpha 1-6PI. This compound is therefore proposed to be an inhibitor of the Dol-P-Man:Man alpha 1-6Man alpha 1-4GlcNa alpha 1-6PI alpha 1-2 mannosyltransferase of the GPI biosynthetic pathway. In living trypanosomes, 4 mM mannosamine had no effect on protein synthesis but reduced the rate of formation of mature glycosylphosphatidylinositol anchor precursors by 80%. This reduction in anchor precursor synthesis was insufficient to prevent the attachment of glycosylphosphatidylinositol anchors to newly synthesized variant surface glycoprotein molecules. These data suggest that the rate of anchor precursor synthesis in the bloodstream form of T. brucei, in contrast to mammalian cells and the procyclic form of T. brucei, is in large excess of the cellular requirements for protein anchorage. PMID- 8226966 TI - The purification and characterization of recombinant yeast dolichyl-phosphate mannose synthase. Site-directed mutagenesis of the putative dolichol recognition sequence. AB - Yeast dolichyl-phosphate-mannose synthase was purified from cultures of Escherichia coli carrying the gene for this enzyme in a high expression vector. The synthase contains a highly conserved hydrophobic amino acid sequence proposed to be involved in the recognition of dolichols (Albright, C. F., Orlean, P., and Robbins, P. W. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 7366-7369) and amino acid residues in this sequence were altered by site-directed mutagenesis. Conservative substitutions had no effect on the affinity of the enzyme for dolichyl-P. The substitution of asparagine for isoleucine at position 253 resulted in higher values for the apparent Km for Dol-P when assayed in detergent solutions, but this substitution had no effect on Km when the enzyme was reconstituted with phosphatidylethanolamine. Enzyme containing a deletion of the entire putative dolichol recognition sequence retained catalytic activity. The apparent Km for Dol-P was increased when this enzyme was assayed in detergent solution but was the same as wild type enzyme when reconstituted in phosphatidylethanolamine. These results suggest that the amino acid composition and sequence of the conserved domain are not critically important for the recognition and binding of Dol-P when the synthase is present in a lipid matrix. PMID- 8226967 TI - A cAMP-regulated RNA-binding protein that interacts with phosphoenolpyruvate carboxykinase (GTP) mRNA. AB - Cyclic-AMP stabilizes phosphoenolpyruvate carboxykinase (GTP) (PEPCK) mRNA against degradation. To investigate the mechanism of this effect, RNA mobility shift assays were used to determine the interaction of cellular proteins with specific domains from the mRNA. We report here the identification of a protein with an affinity for sequences of PEPCK mRNA with a predicted stem-loop structure. RNA-protein complex formation was significantly reduced if the double stranded RNA probe was preheated to 90 degrees C. The RNA-binding protein did not bind to the hairpin structure of poly(rI)-poly (rC), indicating some degree of sequence specificity and that the RNA-binding protein is not the interferon induced double-stranded RNA-activated protein kinase. The binding activity was contained in the cytosolic fraction (100,000 x g) of rat hepatoma FTO-2B cells and was significantly enhanced by high concentrations of KCl. Chromatography on an anion exchanger separated the binding activity from a factor which, upon reconstitution, inhibited the interaction with the RNA probe. Incubation of cells with cAMP resulted in a 3-4-fold decrease in the activity of the RNA-binding protein. An inhibition in complex formation was observed with extracts as early as 60 min after exposure of cells to cAMP. Liver extracts from rats starved for 72 h also had reduced binding activity compared to extracts from fed animals. Cellular extracts treated with alkaline phosphatase exhibited an elevated level of complex formation. An analysis by SDS-polyacrylamide gel electrophoresis of the RNA-protein complex after ultraviolet light cross-linking demonstrated that the RNA-binding protein had a molecular mass of approximately 100 kDa. On the basis of these results, we suggest that liver cells contain a protein whose interaction with PEPCK mRNA is regulated by cAMP-dependent phosphorylation and which may be responsible for the cAMP-mediated control of PEPCK mRNA half-life. PMID- 8226968 TI - Modulation of actin microfilament dynamics and fluid phase pinocytosis by phosphorylation of heat shock protein 27. AB - We recently reported that overexpression of heat shock protein 27 (HSP27) in rodent fibroblasts increases the stability of stress fibers during hyperthermia and partially prevents actin depolymerization during exposure to cytochalasin D (Lavoie, J.N., Gingras-Breton, G., Tanguay, R. M., and Landry, J. (1993) J. Biol. Chem. 268, 3420-3429). Because HSP27 is a ubiquitous target of phosphorylation upon cell stimulation with a variety of growth factors and agents that affect cellular differentiation, we examined the role of HSP27 phosphorylation in regulating actin filament dynamics. Here we show that HSP27 is enriched at the leading edge of polarized fibroblasts. HSP27 is localized in lamellipodia and membrane ruffles where most actin polymerization occurs. We developed Chinese hamster cell lines that constitutively overexpressed either human HSP27 or a nonphosphorylatable mutant form of the protein. Overexpression of HSP27 caused an increased concentration of filamentous actin (F-actin) at the cell cortex and elevated pinocytotic activity. In contrast, overexpression of the non phosphorylatable mutant form of HSP27 reduced cortical F-actin concentration and decreased pinocytosis activity relative to control cells. Mitogenic stimulation of fibroblasts resulted in a rapid polymerization of submembranous actin filaments. HSP27 enhanced growth factor-induced F-actin accumulation, whereas mutant HSP27 exerted a dominant negative effect and inhibited this response to growth factors. Thus, HSP27 is a component of a signal transduction pathway that can regulate microfilament dynamics. PMID- 8226969 TI - Inhibition of basic fibroblast growth factor-induced growth promotion by overexpression of syndecan-1. AB - The expression of syndecan-1, the prototype member of the cell surface proteoglycan family, follows morphogenetic rather than histological boundaries during organ formation. As a heparan sulfate-containing cell surface molecule, syndecan-1 can simultaneously bind various components of the extracellular matrix and members of the heparin-binding growth factors. Indeed, syndecan-1 may act as a co-receptor for basic fibroblast growth factor (bFGF) (Salmivirta, M., Heino, J., and Jalkanen, M. (1992) J. Biol. Chem. 267, 17606-17610), allowing the growth factor to bind the tyrosine kinase bFGF receptor. We have studied the role of syndecan-1 in growth factor response by growing 3T3 cells transfected with syndecan-1 in the presence of bFGF. The enhanced expression of syndecan-1 caused down-regulation of bFGF-induced cell proliferation and, at the same time, enhancement of cell matrix interactions. Thus, the induced expression of the heparan sulfate co-receptor (syndecan-1) may provide a mechanism to restrict FGF action and modulate cell-matrix interactions to maintain co-ordinated growth of cells during organ formation. PMID- 8226970 TI - Structural analysis of the light subunit of the Entamoeba histolytica galactose specific adherence lectin. AB - Adherence of Entamoeba histolytica trophozoites to colonic mucins and resistance to lysis by the membrane attack complex of complement are mediated by a galactose and N-acetyl-D-galactosamine-specific cell-surface lectin. This lectin is a heterodimeric glycoprotein of heavy (170 kDa) and light (35/31 kDa) subunits. In this work, the amino acid sequence and membrane anchor of the light subunit were analyzed. The light subunit cDNA encoded a protein with a calculated molecular mass of 32 kDa containing two potential sites for N-linked glycosylation and putative amino- and carboxyl-terminal signal sequences characteristic of glycosylphosphatidylinositol (GPI)-anchored proteins. No classical carbohydrate binding domains common to C- or S-type eukaryotic lectins were detected by sequence analysis of either the heavy or light subunits, leaving the location of the ligand-binding site of the lectin unknown. Analysis of restriction enzyme digested E. histolytica DNA by Southern blotting was consistent with the presence of more than one light subunit gene. Two light subunit isoforms of 31 and 35 kDa were identified by SDS-polyacrylamide gel electrophoresis analysis of affinity purified lectin, and the isoforms were shown on two-dimensional gel analysis to form distinct 170/35- and 170/31-kDa heterodimers. The amino acid compositions and cyanogen bromide peptide patterns of the two light subunit isoforms were nearly identical. The 35-kDa isoform labeled more efficiently than the 31-kDa isoform with [3H]glucosamine, while only the 31-kDa isoform labeled with [3H]myristate and [3H]palmitate. Nitrous acid deamination released lipid from the 31-kDa isoform, which co-migrated on thin layer chromatography with acylphosphatidylinositol, a component of some GPI anchors. Gas chromatography and mass spectrometry of the deamination product from the 31-kDa subunit identified both myo- and chiro-inositols, supporting the presence of a GPI membrane anchor. The covalent association of a transmembrane protein with a GPI-anchored protein, as suggested by the cDNA sequences of the lectin heavy and light subunits, is novel and suggests unique roles for the two subunits in the pathogenesis of amebiasis. PMID- 8226971 TI - Differences in phosphorylation of formylpeptide and C5a chemoattractant receptors correlate with differences in desensitization. AB - To define the regulation of chemoattractant receptors, epitope-tagged human formyl peptide and C5a receptor cDNAs (ET-FR and ET-C5aR) were stably expressed in rat basophilic leukemia, RBL-2H3 cells. An antibody (12CA5) specific to "ET" was used to immunoprecipitate ET-FR and ET-C5aR. fMLP and C5a caused time- and dose-dependent phosphorylation of their respective receptors. Phosphorylated ET FR migrated as a single broad band between 50 and 70 kDa on SDS-polyacrylamide gel electrophoresis, whereas ET-C5aR exhibited both fast (39-45 kDa) and broadly (39-52 kDa) migrating forms. Fast form phosphorylation alone was observed at low concentrations of C5a (0.001-0.01 microM), or at early times (5-30 s) with a higher concentration of C5a (0.1 microM). Phorbol 12-myristate 13-acetate, thrombin, or antigen caused no phosphorylation of ET-FR but stimulated exclusively fast form phosphorylation of ET-C5aR. The protein kinase C inhibitor staurosporine did not inhibit phosphorylation of ET-FR but blocked the fast migrating component of phosphorylated ET-C5aR. Homologous desensitization correlated with ligand-induced phosphorylation of both receptors. Of note, ET C5aR but not ET-FR underwent heterologous desensitization by antigen, phorbol 12 myristate 13-acetate, and thrombin. The data suggest that protein kinase C mediates heterologous phosphorylation and desensitization of C5aR but not FR, yet, both receptors are homologously desensitized by a staurosporine-resistant kinase. PMID- 8226972 TI - Selective recruitment of masked maternal mRNA from messenger ribonucleoprotein particles containing FRGY2 (mRNP4). AB - In Xenopus, the germ cell-specific Y-box (CTGATTGGCCAA) factor, FRGY2, facilitates in vitro transcription in egg extracts from oocyte-selective promoters containing a Y-box. This same factor is a major component of the messenger ribonucleoprotein (mRNP) storage particles of the oocyte. These particles store maternal mRNAs and inhibit their translation. FRGY2 is identical to mRNP4 and homologous to mRNP3, two previously described oocyte-specific mRNP proteins. We demonstrate that FRGY2 associates with a broad spectrum of mRNAs exhibiting no apparent sequence specificity. These interactions suggest that FRGY2 has a general role in packaging mRNA analogous to that of histone with DNA. All mRNAs examined that accumulate in oocytes appear within these mRNP storage particles, including messages coding for proteins such as FRGY2 and TFIIIA, which are translated in the oocyte. Moreover, we show that mRNAs that are translationally repressed in oocytes, such as messages coding for the proteins histone H1 and FRGY1, accumulate only within the particles. These mRNAs are subsequently recruited from the particles to the ribosomes and utilized for translation during embryogenesis prior to transcriptional activation of the zygotic genome. We propose that the assembly of mRNP storage particles represents a default state and that translational regulation is achieved via specific recruitment of the messages from the mRNP fraction to the ribosomes. PMID- 8226973 TI - Intramitochondrial protein sorting. Isolation and characterization of the yeast MSP1 gene which belongs to a novel family of putative ATPases. AB - Replacement of the presequence of yeast cytochrome c1 by the amino-terminal 61 residues of MAS70, a yeast mitochondrial outer membrane protein, resulted in exclusive localization of the fusion protein (termed the 61-mC1 protein) to the outer membrane. When a cytochrome c1-deficient yeast strain was transformed with a plasmid encoding the fusion protein, the cells could not grow on nonfermentable carbon sources such as glycerol. We isolated a novel yeast gene MSP1 (mitochondrial sorting of proteins) whose overexpression causes mislocalization of the 61mC1 fusion protein to the inner membrane, probably via the intermembrane space, and thereby allows the host cells to grow on glycerol. The predicted MSP1 protein (MSP1) is a hydrophilic 40-kDa polypeptide containing a putative membrane spanning domain near the amino terminus. Further sequence analyses revealed that MSP1 is a member of a novel family of putative ATPases which share a highly conserved domain of about 185 amino acid residues, including a consensus motif for a nucleotide binding site. MSP1 was found to be an intrinsic mitochondrial outer membrane protein of an apparent molecular mass of 40 kDa with a large domain facing to the cytosol. The MSP1 gene is not essential for the cell growth either on fermentable or nonfermentable carbon sources. PMID- 8226974 TI - Defective processing of the insulin receptor in an endoprotease-deficient Chinese hamster cell strain is corrected by expression of mouse furin. AB - Characterization of an endoprotease-deficient mutant Chinese hamster ovary (CHO) cell, designated RPE.40, revealed that it bound less than 10% as much insulin as did its parent, CHO-K1. We examined processing of the endogenous insulin receptor in CHO-K1 and RPE.40 cells, and processing of the human insulin receptor expressed in these cells. RPE.40 cells did not process the endogenous insulin proreceptor to its subunit forms, and processed the human insulin proreceptor inefficiently. Accumulation of the proreceptor form of the insulin receptor was seen in both cases. Furin is a mammalian endoprotease that cleaves proproteins at a consensus sequence of basic amino acids found in the insulin proreceptor. Expression of mouse furin in RPE.40 cells restored normal processing of the endogenous and the human insulin receptor in these cells. In addition, expression of mouse furin corrected the reduced binding of insulin in RPE.40 cells, indicating that receptor function as well as processing was restored. PMID- 8226975 TI - Distinct dimerization domains provide antagonist pathways for thyroid hormone receptor action. AB - Transcriptional regulation by thyroid hormone is mediated through its nuclear receptors, which bind to target response elements as homodimers or as heterodimers with proteins such as retinoid X receptors (RXR). Thyroid hormone response elements exhibit remarkable flexibility in that the receptor binding half-sites can be arranged as direct repeats, inverted repeats, or everted repeats. We report that a limited region at the carboxyl-terminal end of the thyroid hormone receptor differentially contributes to the formation of receptor homo- and heterodimers. The functionally inactive thyroid hormone receptor splicing variant alpha 2, which is altered at the juncture of the homo- and heterodimerization domains, cannot form homodimers. However, alpha 2 can form a heterodimer when bound to half-sites arranged as a direct repeat spaced by 4 base pairs (DR4), but not with other arrangements of response element half-sites. The alpha 2-RXR heterodimer strongly inhibits wild type receptor function mediated by the DR4 element, suggesting that the alpha 2 isoform modulates thyroid hormone action by binding as an antagonist to a subset of response elements. PMID- 8226976 TI - Multiple mRNA isoforms encoding the mouse cardiac Kv1-5 delayed rectifier K+ channel. AB - The mouse Kv1-5 K+ channel cDNA has been cloned from heart. This channel was highly expressed in heart and, to a lesser extent, in other tissues, including brain and thymus. Two alternatively spliced isoforms were found. The longer form encoded a 602-amino acid protein, while in the short form (Kv1-5 delta 5'), the first 200 amino acids lying upstream the transmembrane segment S1 were deleted. RNase protection experiments showed that both Kv1-5 mRNA isoforms are present in the mouse tissues examined, the longer form being predominant. The short mRNA (Kv1-5 delta 5') arose by an unusual splicing event within the exonic sequence. An additional short cDNA clone (Kv1-5 delta 3') that codes for a carboxyl terminal truncated protein has been isolated. The gene coding sequence contained a single exon and has been mapped on human chromosome 12 (p13) and on mouse chromosome 6 (band F). Expression in Xenopus oocytes revealed that the long (Kv1 5) and the amino-terminal deleted (Kv1-5 delta 5') isoforms elicited similar K+ currents with a drastically decreased efficacy for Kv1-5 delta 5'. The carboxyl terminal truncated Kv1-5 delta 3' clone was not functional but inhibited the expression of the long isoform. PMID- 8226977 TI - Molecular cloning of trkE, a novel trk-related putative tyrosine kinase receptor isolated from normal human keratinocytes and widely expressed by normal human tissues. AB - We have identified and cloned a new member of the trk gene family, termed trkE, which generates a 3.9-kilobase (kb) transcript in normal human keratinocytes and in a variety of normal human tissues, but not in liver. Albeit at low level, trkE transcript is expressed also by PC12 cells. The open reading frame codes for a polypeptide of 876 amino acids exhibiting the classic features of cell surface tyrosine protein kinases. trkE catalytic domain is 41% identical to trkA and shows several features unique to the trk gene family. Its extracellular domain does not show significant homology to any known proteins. trkE is the first member of this gene family found abundantly and widely expressed in normal human tissues. Several lines of evidence suggest that NGF is also the ligand for trkE; (i) normal human keratinocytes bind NGF with high affinity, (ii) NGF stimulates keratinocyte growth in an autocrine fashion, (iii) NGF exerts its biological effect on keratinocytes through the stimulation of a trk-specific tyrosine kinase, and (iv) keratinocytes lack trkA but do express large amount of trkE. trkE might also be the NGF receptor by other human peripheral tissues, such as pancreatic islets, and might represent a non-neuronal receptor for this ligand. PMID- 8226978 TI - Molecular cloning and characterization of PKC iota, an atypical isoform of protein kinase C derived from insulin-secreting cells. AB - The protein kinase C (PKC) family of serine-threonine kinases comprises at least eight members. These are differentially expressed, show varying affinities for activators such as Ca2+ and lipid species, and are therefore thought to play wide ranging roles in the regulation of such cellular processes as differentiation, growth, and secretion. The aim of this study was to identify new PKC isoforms in the insulin-secreting cell line RINm5F that might be activated by the alterations in lipid metabolism that accompany nutrient-stimulated insulin release. Fragments of cDNA, derived from RINm5F cell mRNA, were amplified by the polymerase chain reaction using degenerate oligonucleotide primers corresponding to highly conserved regions in the catalytic domains of all known PKCs. A novel sequence generated by this approach was subsequently used to screen cDNA libraries. The entire 587-amino acid coding region of a new PKC isoform, PKC iota, was deduced from two overlapping clones isolated from a human kidney cDNA library. The amino acid sequence of PKC iota showed greatest homology to PKC zeta, with 72% identity overall rising to 84% in the catalytic domain. In contrast, the homology of PKC iota to the other isoforms was less pronounced, with < 53% identity even in the highly conserved catalytic region. Further similarities between PKC zeta and PKC iota included a highly conserved pseudosubstrate sequence, the absence of an apparent Ca(2+)-binding region, and the presence of only one cysteine-rich, zinc finger-like domain. Northern blot analysis, using the full-length PKC iota clone as a probe, revealed a single 4.6-kilobase transcript present predominantly in lung and brain, but also expressed at lower levels in many tissues including pancreatic islets. In CHO-K1 cells stably expressing the PKC iota cDNA under the human beta-actin promoter, the protein was detected as a 65-kDa band by Western blotting using an antibody to the COOH terminus of PKC zeta (conserved in PKC iota). Extracts of transfected CHO-K1 cells also displayed a significantly increased kinase activity using myelin basic protein as a substrate. The results suggest that PKC iota should be included in the atypical subgroup of PKCs whose definitive member is PKC zeta. As such, PKC iota is unlikely to be activated by the diacylglycerol that is derived from phosphoinositide hydrolysis, but might be a target for novel lipid activators that are elevated during nutrient-stimulated insulin secretion. PMID- 8226979 TI - Structure and function of C-CAM1. The first immunoglobulin domain is required for intercellular adhesion. AB - Cell-CAM105 proteins (also called C-CAM) are epithelial cell adhesion molecules of the immunoglobulin (Ig) superfamily. The sequences of C-CAM are highly homologous to those of human carcinoembryonic antigen (CEA)-family proteins. In previous studies using baculoviral vectors, we showed that expression of the L form cell-CAM105 (also called C-CAM1) in insect cells resulted in cell aggregation (Cheung, P. H., Thompson, N. L., Earley, K., Culic, O., Hixson, D., and Lin, S. H. (1993) J. Biol. Chem. 268, 6139-6146). This result indicates that the insect-cell system is suitable for studying the adhesion function of C-CAM. Since C-CAM1 contains four extracellular Ig-domains, the structural features directly responsible for C-CAM1 adhesion function were investigated by site directed deletion and expression in the baculovirus/insect cell system. Results from these studies indicated that the first Ig domain located in the NH2-terminal of C-CAM plays a crucial role in intercellular adhesion. Site-directed deletion producing mutants lacking the second, third, or fourth Ig domains had no effect on the adhesion function. In addition, adhesion function was retained when both the third and fourth Ig domains were deleted, although the adhesion activity was reduced to half that in control cells. However, simultaneous deletion of the second, third, and fourth domains abolished adhesion, suggesting that these domains affect the accessibility of the binding site localized in the first domain. In our previous studies, we showed that the cytoplasmic domains of C-CAM play a significant role in the isoforms' adhesion activity since expression of a C-CAM isoform containing only 6 instead of 71 amino acids intracellularly failed to show the adhesion phenotype (Cheung, P. H., Culic, O., Qiu, Y., Earley, K., Thompson, N., Hixson, D. C., and Lin, S.-H. (1993) Biochem. J. 295, in press). These results together suggest that both the cytoplasmic domain and the first N terminal Ig-like domain are required for C-CAM-mediated cell adhesion activity. PMID- 8226980 TI - Purification and characterization of 2,4-dichlorophenoxyacetate/alpha ketoglutarate dioxygenase. AB - The Alcaligenes eutrophus 2,4-dichlorophenoxyacetate/alpha-ketoglutarate dioxygenase, encoded by the tfdA gene of plasmid pJP4, is an Fe(II)-dependent enzyme that catalyzes the conversion of 2,4-dichlorophenoxyacetate to 2,4 dichlorophenol and glyoxylate concomitant with the decomposition of alpha ketoglutarate to form succinate and carbon dioxide (Fukumori, F., and Hausinger, R. P. (1993) J. Bacteriol. 175, 2083-2086). Using recombinant Escherichia coli cells that overexpress the tfdA gene, the thermolabile enzyme (stable only up to 30 degrees C) was purified to apparent homogeneity (specific activity of 16.9 mumol of substrate converted min-1 mg of protein-1) by a simple two-step procedure. The native protein has an apparent M(r) of 50,000 +/- 2,500, consistent with a homodimeric structure. Ferrous ion is absolutely required for activity and cannot be replaced by several other divalent cations tested. Ascorbic acid stimulates dioxygenase activity and reduces the rate of enzyme inactivation by a metal ion-mediated process. The enzyme exhibits maximum activity at pH 6.5-7, however, it is stable over a pH range of 6.5-11. Although capable of hydroxylating a wide range of phenoxyacetates and related compounds, the enzyme exhibits the greatest affinity (Km 17.5 +/- 1.0 microM) and highest catalytic efficiency for 2,4-dichlorophenoxyacetate. Similarly, alpha ketoglutarate is the preferred co-substrate (Km 3.20 +/- 0.54 microM) for the enzyme, but it can utilize a range of other alpha-ketoacids with lower efficiency. Results from chemical modification studies are consistent with the presence of multiple essential histidine residues in the enzyme. PMID- 8226981 TI - Dipeptide processing activates recombinant human prochymase. AB - Human chymase (h-chymase) is a serine protease that efficiently converts angiotensin I to II. Its structure and homology to other serine proteases suggest that it is synthesized as a zymogen, and is processed to the active form by cleavage of a 19-residue signal peptide and of a dipeptide pro-segment. To evaluate maturational processing of this enzyme, the proteins encoded by three h chymase cDNA constructs (wild-type, lacking the pro- or lacking the prepro segment) were characterized after expression in COS-1 cells. These recombinant proteins were not catalytically active. Purification and NH2-terminal sequence analysis of the protein expressed from the wild-type construct revealed processing to the proenzyme. Prochymase activation was achieved by incubation with a B-cell lymphoma homogenate, which apparently contains a heterologous processing enzyme sensitive to thiol protease inhibitors. NH2-terminal sequence analysis of the activated h-chymase revealed cleavage of the pro-segment, and its biochemical characteristics were identical to those of native h-chymase purified from the myocardium. These findings indicate that processing of the dipeptide pro segment is necessary and sufficient for activation of human chymase. Such processing is probably also required for the activation of related serine proteases, e.g., cathepsin G, which have homologous dipeptide pro-segments. PMID- 8226982 TI - Threonine 204 of the chaperone protein Hsc70 influences the structure of the active site, but is not essential for ATP hydrolysis. AB - The chaperone protein Hsc70 is an ATPase of unknown mechanism, although the crystal structure of the 44-kDa ATPase domain has been solved. This structure shows that the hydroxyl of threonine 204 is located close to the gamma-phosphate of ATP, in a position where it might be an intermediate phosphate acceptor in the hydrolysis reaction. We made two point mutations at residue 204 of Hsc70, threonine to valine (T204V) and threonine to glutamic acid (T204E). The wild-type ATPase domain had a Km for ATP of approximately 1 microM; the mutants had Km values of approximately 90 microM. The kcat values for the mutant proteins were also increased. After crystallization, the structures of the T204V and T204E proteins were solved and refined with data to 2.3- and 2.4-A resolution, respectively. The overall tertiary structure of the mutants showed little change from the wild type; however, significant changes were observed in the active site. Analysis of the structures suggested possible reasons for the changes in kinetic constants. Threonine 204 does not seem to be an obligatory intermediate phosphate acceptor in the hydrolysis reaction since the mutants retained appreciable ATPase activity. PMID- 8226983 TI - The importance of specific gamma-carboxyglutamic acid residues in prothrombin. Evaluation by site-specific mutagenesis. AB - Prothrombin has 10 gamma-carboxyglutamic acid residues which are essential for the metal ion binding properties and membrane binding function of the protein. To assess the importance of each gamma-carboxyglutamic acid residue we made, by site directed mutagenesis, a series of mutant human prothrombins each with a single glutamic acid to aspartic acid substitution at positions 6, 7, 14, 16, 19, 20, 25, 26, 29, or 32 which are gamma-carboxylated in native prothrombin. Along with wild-type prothrombin, the prothrombin mutants were expressed in Chinese hamster ovary cells, purified by immunoaffinity chromatography using polyclonal anti prothrombin antibodies, and shown by amino acid analysis to contain the expected number of gamma-carboxyglutamic acid residues. Only substitution of gamma carboxyglutamic acid 6 with aspartic acid yields a protein with procoagulant activity, affinity for phospholipid and KM(app) for prothrombinase indistinguishable from wild-type prothrombin. In contrast, the conservative gamma carboxyglutamic acid to aspartic acid mutation at positions 16, 26, or 29 results in proteins with little or no procoagulant activity, Kd(app) for binding to phospholipid at least 200-fold higher than wild-type prothrombin and a KM(app) for interaction with the prothrombinase complex nearly 100-fold higher than wild type prothrombin. The mutations at residues 7, 14, 19, 20, 25, or 32 yielded proteins with intermediate procoagulant activities, between 15 and 52% of wild type prothrombin. These data have been interpreted to suggest that there are certain gamma-carboxyglutamic acid residues which are important to maintaining the basic structure of the calcium-liganded Gla domain while other gamma carboxyglutamic acid residues subserve other functions including membrane binding and interdomain interactions. PMID- 8226984 TI - Yeast TKL1 gene encodes a transketolase that is required for efficient glycolysis and biosynthesis of aromatic amino acids. AB - A gene encoding transketolase, TKL1, was cloned from the budding yeast Saccharomyces cerevisiae using oligonucleotide primers derived from protein sequence data. The TKL1 sequence predicts a 74-kDa polypeptide which is related to other transketolases. A sequence comparison revealed that the transketolases can be subdivided into three evolutionary branches. We also found that the transketolases are related to another vitamin B1-dependent enzyme: the E1 subunit of pyruvate dehydrogenase from Escherichia coli. Gene disruption and overexpression experiments were used to investigate the function of transketolase in yeast. We found that growth on fermentable carbon sources, but not on gluconeogenic carbon sources, is reduced in cells disrupted for TKL1. This suggests that the glycolytic efficiency is impaired. Growth on fermentable carbon sources is also reduced in cells that overexpress TKL1. Finally, we found that cells disrupted for TKL1 are unable to grow in the absence of aromatic amino acids. This is most likely due to the fact that transketolase is required for the synthesis of erythrose-4-P, a precursor of the aromatic amino acids. PMID- 8226985 TI - Expression of platelet-derived growth factor beta receptor on human monocyte derived macrophages and effects of platelet-derived growth factor BB dimer on the cellular function. AB - Platelet-derived growth factor (PDGF) plays an important role in the process of atherosclerosis which is characterized by the presence of macrophage-derived foam cells. In the present study, the induction of the mRNA of PDGF-beta receptor was demonstrated during cell differentiation of human monocyte-macrophages, whereas no mRNA was detected in the cells during the early days of culture. Flow cytometry analysis using antibodies specific for PDGF-beta receptor and CD14 showed the presence of both PDGF-beta receptor and CD14 on human monocyte-derived macrophages, whereas no PDGF-beta receptor was detected on human monocytes 4 h after cell adhesion to a culture dish. In the binding assay of PDGF-BB on human monocyte-derived macrophages, a saturable and high affinity binding site with Kd of 27.5 pM and Bmax of 23.3 fmol/mg of cell protein was demonstrated. When human monocytes were cultured in the presence of the protein kinase C inhibitor staurosporine, PDGF-beta receptor induction was inhibited, and tetradecanoylphorbol acetate enhanced PDGF-beta receptor expression in human monocyte-derived macrophages, indicating that PDGF-beta receptor expression is associated with maturation and differentiation of monocyte-macrophages through the activation of protein kinase C. In response to PDGF-BB homodimer, PDGF-beta receptor was phosphorylated, and thymidine uptake and inositol trisphosphate production were stimulated in monocyte-derived macrophages. Furthermore, PDGF-BB suppressed the production of macrophages colony-stimulating factor in macrophages. The expression of PDGF-beta receptor on human monocyte-derived macrophages suggests that PDGF influences the process of atherosclerosis by regulating the function of macrophages as well as smooth muscle cells in the vascular wall. PMID- 8226986 TI - Molecular cloning of fish Pit-1 cDNA and its functional binding to promoter of gene expressed in the pituitary. AB - Pit-1 is a pituitary-specific transcription factor responsible for activating growth hormone (GH) and prolactin genes. Here, we describe the isolation of a rainbow trout cDNA clone that contains the entire Pit-1 coding region. The deduced amino acid sequence contains 358 residues, encoding a 39-kDa protein. Comparison of the protein sequences of the rainbow trout and rat Pit-1 shows that the 160 residue POU domain at the C terminus is highly conserved (86% identical). However, homology is much weaker in the N-terminal region (56% identical), and the rainbow trout Pit-1 contains segments of 29 and 33 amino acids that are not present in rat Pit-1. The protein produced by expression of rainbow trout Pit-1 cDNA in Escherichia coli binds specifically to at least four sites in the rainbow trout GH gene promoter. Moreover, we demonstrate that the promoter region of salmon somatolactin gene, which belongs to the GH prolactin gene family and is also expressed specifically in the pituitary, has at least five rainbow trout Pit 1 binding sites. The consensus sequence of these binding sites closely matches the 9-base pair motif, (T/A)(T/A)TATNCAT, recognized by rat Pit-1. Rainbow trout Pit-1 specifically activates rainbow trout GH promoter fusion gene expression, confirming the ability of Pit-1 to bind in a transcriptionally active conformation in GH gene promoter. PMID- 8226987 TI - Microtubule-associated protein tau. Abnormal phosphorylation of a non-paired helical filament pool in Alzheimer disease. AB - The major protein subunit of the paired helical filaments (PHF) of Alzheimer disease (AD) is the microtubule-associated protein tau. Tau is a family of phosphopolypeptides that are abnormally phosphorylated in PHF. In this study, a non-PHF pool of tau abnormally phosphorylated at Ser-199/202, and tau not phosphorylated at this site (AD P-tau and AD tau, respectively) were isolated from the 27,000 x g to 200,000 x g fraction of AD brain homogenate by extraction in 8 M urea, followed by dialysis against Tris buffer. AD P-tau and AD tau were further purified and separated from each other by acid precipitation, glial fibrillary acidic protein affinity chromatography, and phosphocellulose chromatography. The resulting AD P-tau and AD tau preparations were free of cytoskeletal proteins, ubiquitin, and beta-amyloid peptide. Immunochemical and morphological analysis of AD P-tau preparations revealed that most of the protein was of non-PHF origin. The AD P-tau was about 3-4-fold (approximately 8 mol P04/mol protein, M(r) 41,318) more phosphorylated than cytosolic tau from AD and control brains. Unlike PHF, the AD P-tau lacked ubiquitin. In AD brain the levels of cytosolic tau were about half of those in control aged cases. These findings suggest that the abnormal phosphorylation of tau in AD occurs in the cytosol. PMID- 8226988 TI - Analysis of the mammalian gadd45 gene and its response to DNA damage. AB - The gadd45 gene is transcriptionally activated through at least two different mechanisms; one following treatment with base-damaging agents such as methylmethane sulfonate and UV radiation and the other following ionizing radiation. To investigate the sequences involved in induction of gadd45 by agents producing high levels of base damage, the hamster, human, and mouse genes were sequenced. Comparison of these sequences revealed a high level of conservation between species of 1500 base pairs of the proximal promoter and 700 base pairs within the third intron. However, in the promoter regions, there was no conservation between species of any transcription factor binding sites known to confer DNA damage responsiveness. The promoter of the hamster gene was inducible by base-damaging agents in both rodent and human cell lines and the human gene was inducible in a rodent cell line. This indicates that both sequence elements in the gadd45 promoter and factors binding to these sites are conserved in mammalian cells. Deletion analysis of the hamster promoter did not reveal any specific sequence which conferred damage inducibility and the maximal response required a large portion of the promoter. The hamster promoter was not inducible by ionizing radiation, suggesting that sequences outside the promoter region used, such as a p53 binding site in the third intron, are necessary. The human GADD45 gene was mapped to chromosome 1p31.1-31.2. PMID- 8226989 TI - The purification and characterization of an extremely thermostable alpha-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus. AB - The alpha-amylase from Pyrococcus furiosus, a hyperthermophilic archaebacterium, has been purified to homogeneity. The enzyme is a homodimer with a subunit molecular mass of 66 kDa. The isoelectric point is 4.3. The enzyme displays optimal activity, with substantial thermal stability, at 100 degrees C, with the onset of activity at approximately 40 degrees C. Unlike mesophilic alpha-amylases there is no dependence on Ca2+ for activity or thermostability. The enzyme displays a broad range of substrate specificity, with the capacity to hydrolyze carbohydrates as simple as maltotriose. No subtrate binding occurs below the temperature threshold of activity, and a decrease in Km accompanies an increase in temperature. Except for a decrease in Asp and an increase in Glu, the amino acid composition does not confirm previously defined trends in thermal adaption. Fourth derivative UV spectroscopy and intrinsic fluorescence measurements detected no temperature-dependent structural reorganization. Hydrogen exchange results indicate that the molecule is rigid, with only a slight increase in conformational flexibility at elevated temperature. Scanning microcalorimetry detected no considerable change in the heat capacity function, at the pH of optimal activity, within the temperature range in which activity is induced. The heat absorption peak due to denaturation, under these conditions, occurred within the temperature range of 90-120 degrees C. When the pH was increased, a change in the shape of the heat absorption peak was observed, which when analyzed thermodynamically shows that the process of heat denaturation is complex, and includes at least three stages, indicating that the protein structure consists of three domains. At temperatures below 90 degrees C no excess heat absorption or change in the CD spectra were observed which could be associated with the cooperative conformational transition of the protein. According to the thermodynamic characteristics of the heat denaturation, the cold denaturation of this protein can be expected only at -3 degrees C. Therefore, the observed inactivation of this enzyme is not caused by the cooperative change of its tertiary structure. It can be associated only with the gradual changes of protein domain interaction. PMID- 8226990 TI - Alpha-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus. Cloning and sequencing of the gene and expression in Escherichia coli. AB - A gene encoding a highly thermostable alpha-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus was cloned and expressed in Escherichia coli. The nucleotide sequence of the gene predicts a 649-amino acid protein with a calculated molecular mass of 76.3 kDa, which corresponds well with the value obtained from purified enzyme using denaturing polyacrylamide gel electrophoresis. The NH2 terminus of the deduced amino acid sequence corresponds precisely to that obtained from the purified enzyme, excluding the NH2-terminal methionine. The amylase expressed in E. coli exhibits temperature-dependent activation characteristic of of the original enzyme from P. furiosus, but has a higher apparent molecular weight which is attributed to the improper formation of the native quaternary structure. No homology was found with previously characterized promotor or termination sequences. The deduced amino acid sequence displayed strong homology to the alpha-amylase A of Dictyoglomus thermophilum, an obligately anaerobic, extremely thermophilic bacterium. Evolutionary implications of this homology are discussed. PMID- 8226991 TI - Evolutionary conservation of synaptosome-associated protein 25 kDa (SNAP-25) shown by Drosophila and Torpedo cDNA clones. AB - The neuron-specific proteins SNAP-25 (synaptosome-associated protein 25 kDa), synaptobrevin and syntaxin, are localized to presynaptic terminals in mammals and have been found to associate with proteins involved in vesicle docking and membrane fusion. We describe here SNAP-25 cDNA clones from the fruit fly Drosophila melanogaster and the ray Torpedo marmorata. In situ hybridization showed that SNAP-25 mRNA is exclusively found in brain and ganglia in Drosophila with a pattern suggesting expression in most neurons. The Drosophila and Torpedo proteins show 61 and 81% amino acid identity to mouse SNAP-25, a degree of conservation similar to that previously reported for synaptobrevin. None of the SNAP-25 sequences has a membrane-spanning region, but all contain a cluster of cysteine residues that can be palmitoylated for membrane attachment. SNAP-25 displays sequence similarity to syntaxin A and B. These data show that SNAP-25 and synaptobrevin, which are both implicated in vesicle docking and/or membrane fusion, have both been highly conserved during evolution. This supports the existence of a basic molecular machinery for synaptic vesicle docking in vertebrate and invertebrate synapses. PMID- 8226992 TI - Phosphorylation inhibits the DNA-binding activity of MyoD homodimers but not MyoD E12 heterodimers. AB - MyoD is a member of the basic helix-loop-helix (bHLH) family of muscle gene regulatory proteins that includes myogenin, myf-5, and MRF4. These proteins have been shown to heterodimerize with E2A bHLH proteins, E12/E47, and to bind to a consensus sequence known as an E-box, CANNTG, the target for transcriptional activation by these myogenic regulators. MyoD is also a phosphorylated nuclear protein that is present in muscle cells prior to the transcriptional activation of the muscle-specific genes, many of which contain E-box elements in their regulatory regions. Here we report that phosphorylated chicken MyoD, called CMD1, produced in sf9 cells using the baculovirus system, is qualitatively similar to CMD1 isolated by immunoaffinity purification from primary cultures of embryonic chick breast muscle. Functional analysis of phosphorylated and dephosphorylated CMD1 produced in sf9 cells indicates that, in the presence of magnesium, DNA binding of phosphorylated CMD1 is inhibited whereas binding in association with E12 is not affected. However, CMD1 binding alone is equally efficient when either EDTA is added in excess or dephosphorylated or bacterially expressed CMD1 is used in the assay. Our results suggest that cellular phosphorylation changes the CMD1 homodimer-heterodimer equilibrium which, in turn, modulates and/or eliminates binding site competition between CMD1 homodimers and CMD1/E-protein heterodimers in the cell. PMID- 8226993 TI - A highly conserved region of human erythrocyte ankyrin contains the capacity to bind spectrin. AB - Ankyrin has a spectrin-binding region within a central 62-kDa chymotryptic peptide. We examined the spectrin binding ability of a series of smaller ankyrin fragments and recombinant peptides within the 62-kDa domain using a ligand blot assay. The smallest proteolytic fragment that bound was a 12-kDa tryptic peptide starting at amino acid 1068. Peptides containing this region expressed as glutathione S-transferase fusion products also bound spectrin and suggested that residues 1101-1192 were important. In contrast, a fusion protein containing residues 826-898 did not bind spectrin, a surprising finding since this region is known to influence binding affinity. Proteins that bound spectrin on ligand blots also competed for binding in solution, but did so with one-tenth the affinity of the native peptide. Comparing the 62-kDa domains of erythrocyte and brain ankyrins (species that bind spectrin but with 10-fold differences in affinity), the NH2-terminal regions are 0-40% identical, while the regions (1136-1160) common to all binding peptides are 80-90% identical. We hypothesize that the highly conserved region contains an important spectrin-binding site, while the poorly conserved region controls the binding affinity. We speculate that this unique NH2-terminal region is what gives different members of the ankyrin family their signature set of affinities, and accordingly their distinctive cellular localization. PMID- 8226994 TI - Cross-linking of Fc gamma receptor I (Fc gamma RI) and receptor II (Fc gamma RII) on monocytic cells activates a signal transduction pathway common to both Fc receptors that involves the stimulation of p72 Syk protein tyrosine kinase. AB - Stimulation of the human monocytic cell line THP-1 by cross-linking either Fc gamma receptor I (Fc gamma RI) or Fc gamma receptor II (Fc gamma RII) gave rise to the rapid phosphorylation of multiple intracellular proteins. The pattern of proteins that were phosphorylated appeared to be identical. Analysis of these proteins by specific immunoprecipitation indicated that stimulation through either receptor did indeed give rise to phosphorylation of the same set of proteins. These included: Fc gamma RII, phospholipase C (PLC) gamma 1, PLC gamma 2, Vav, GAP, and a protein that co-precipitated with the Fc gamma receptors and migrated with a molecular weight of about 70,000. Co-cross-linking an F(ab')2 anti-CD45 monoclonal antibody together with monoclonal antibodies to either of the Fc gamma receptors inhibited phosphorylation of all these proteins. Analysis of the tyrosine kinases in the cells revealed that both receptors stimulated the phosphorylation and activation of a kinase recognized by antibodies to Syk. Furthermore, the Syk kinase became associated with the Fc gamma RII following receptor cross-linking. These data indicate that although the two Fc gamma receptors have different cytoplasmic tails, they are coupled to the same signal transduction cascade that is regulated by CD45 and involves the activation of Syk. PMID- 8226995 TI - The Rab3A GTPase interacts with multiple factors through the same effector domain. Mutational analysis of cross-linking of Rab3A to a putative target protein. AB - Rab3A/smg25A is a small Ras-like guanine nucleotide binding protein implicated in the control of regulated secretion from cells. Rab3A is approximately 30% cytosolic and 70% associated with the membranes of secretory vesicles. It cross links specifically to a rat brain membrane protein of about 85 kilodaltons (p85). To identify epitopes on Rab3A that are important for its interaction with this putative target protein, we have determined the effects of point mutations on the cross-linking efficiency of Rab3A to p85. Rab3A, which was preincubated with a non-hydrolyzable analog of GTP, cross-linked more efficiently to p85 than did Rab3A-GDP. Rab3A mutants that had decreased nucleotide binding also exhibited poor cross-linking to p85. Mutations in the effector domain, a site important for the interaction of Rab3A with its guanine nucleotide releasing factor, guanine nucleotide dissociation inhibitor, and GTPase-activating protein, eliminated the ability of Rab3A to cross-link to p85. However, short peptides corresponding to the effector domain did not reduce cross-linking efficiency when present at a concentration of 50 microM. PMID- 8226996 TI - Differential expression of S1 and elongation factor-1 alpha during rat development. AB - Elongation factor-1 alpha (EF-1 alpha) is a highly conserved protein functioning in peptide elongation during translation. A cDNA, S1, was isolated; its deduced amino acid sequence shares high similarity with mammalian EF-1 alpha s (92%). While EF-1 alpha is present in all tissues, S1 mRNA can only be detected in brain, heart, and muscle. We report here that the retropseudogene phenomenon is attributable to EF-1 alpha and not S1, the latter being represented by a single copy in the rat genome. The S1 steady-state mRNA levels are consistently higher than EF-1 alpha in S1-positive tissues. S1 mRNA can only be detected late during brain, heart, and muscle development in vivo and increases to a plateau in early postnatal life. In a cultured muscle system, S1 expression is dependent upon the formation of myotubes, although the accumulation of S1 mRNA is significantly lower than that observed in adult skeletal muscle. EF-1 alpha mRNA levels are down-regulated during brain, heart, and muscle development, but stay relatively steady in liver. We show here that EF-1 alpha and S1 are differentially expressed during rat development and that the activation of S1 gene expression is subsequent to the terminal differentiation process in brain, heart, and muscle. PMID- 8226997 TI - Distinct TATA motifs regulate differential expression of human metallothionein I genes MT-IF and MT-IG. AB - In this report, we have measured the cadmium (Cd2+)-induced expression of all known metallothionein I (MT-I) mRNAs in a human hepatoma cell line, Hep G2. Among the human MT-I gene family promoters, marked sequence conservation exists; despite this, the mRNA accumulation level for each species was found to be quite unique. This differential Cd2+ induction of MT-I family members provides an ideal opportunity to assess whether the characteristic response results from subtle isoform-specific variations in promoter structure. Accordingly, we have examined the mechanism for differential expression of two isoforms, MT-IG and MT-IF, by transient transfection into Hep G2 cells. In the presence of Cd2+, MT-IG promoter activity and endogenous mRNA level were, respectively, 4.7- and 3-fold greater than those of MT-IF. This close correlation between promoter activity and mRNA accumulation strongly suggests that differential expression occurs at the level of transcription. The difference in Cd(2+)-stimulated activity was found to be conferred by 240- and 243-base pair promoter fragments spanning nucleotides -174 to +66 and -172 to +71 of the MT-IG and MT-IF genes, respectively. One of the most striking nonhomologies between the promoters is a single A (TATAAA) to C (TATCAA) transversion in the TATA motifs of MT-IG and MT-IF genes, respectively. To determine whether such a subtle change in the TATA motif could account for the marked differences in promoter function, we constructed MT-IG-TATCA and MT-IF TATAA promoters and measured their activities in transient transfection and cell free transcription assays. Results of both assays showed a profound difference between the two motifs that paralleled the difference in Cd(2+)-stimulated MT-IG and MT-IF mRNA levels. In summary, we have shown that differential regulation of two MT-I promoters is primarily due to a single base alteration in their TATA motifs. PMID- 8226998 TI - Post-translational processing of Schizosaccharomyces pombe YPT5 protein. In vitro and in vivo analysis of processing mutants. AB - SpYPT5p is a member of the rab/YPT small GTP-binding protein family, which is believed to be involved in the regulation of intracellular trafficking. The protein sequence terminates with a CXC motif, and in our previous report (Newman, C. M. H., Giannakouros, T., Hancock, J. F., Fawell, E. H., Armstrong, J., and Magee, A. I. (1992) J. Biol. Chem. 267, 11329-11336) we have shown that SpYPT5p is prenylated both in vivo and in vitro, where geranylgeranylation was confirmed, and carboxyl-methylated. In order to dissect the role of prenylation of each cysteine, we have generated C-terminal mutants where either one or both cysteine(s) were replaced by serine and expressed them in vitro in reticulocyte lysates and in vivo in transfected COS cells. Our results suggest that both cysteines of the CXC motif are prenylated but that the rate of prenylation of the two cysteines is different. The upstream cysteine was found to be preferentially prenylated in reticulocyte lysates unless cytosol from COS cells was added. A separate activity could therefore be required for prenylation of the second cysteine, or the presence of an additional factor is needed to allow accumulation of doubly prenylated SpYPT5p. However, the modification of the upstream cysteine is not a prerequisite for the prenylation of the other. Furthermore, gene replacement in Schizosaccharomyces pombe revealed that each cysteine of the CXC motif can individually support function. Carboxyl methylation occurred only on protein which had been prenylated on the C-terminal cysteine and was required for efficient membrane binding in vitro. PMID- 8226999 TI - Structure-function relationship of the small GTPase rab5. AB - Overexpression of rab5 via a Sindbis virus vector resulted in a 2-3-fold stimulation of horseradish peroxidase uptake in BHK-21 cells. Based on this functional assay of rab5 activity, we conducted extensive mutational analysis of the structure-function relationship of rab5. A total of 21 deletion and substitution mutations were created and their effects on rab5 activity were examined. Deletion of the entire C-terminal tetrapeptide motif CCSN abolished rab5 activity. A mutant with the last three residues deleted, however, showed residual rab5 activity. Truncation of only two residues from the C terminus had no effect on the biological activity of rab5. A mutant containing a 4-residue deletion from the N terminus retained full activity in comparison with wild-type rab5. N-terminal deletion of 19 residues only partially blocked rab5 activity. Substitution mutations in the guanine nucleotide binding motifs showed dramatic effects on rab5 function. In addition to the previously reported N133I mutation, the S34N mutation also resulted in a guanine nucleotide binding defective form that was a dominant inhibitor of endogenous rab5 activity. The Q79L mutation (the ras equivalent Q61L decreases intrinsic and GTPase-activating protein-stimulated GTPase activities), however, had no effect on rab5 activity. The S35N mutation, which is immediately downstream of the first GTP/GDP binding motif, decreased guanine nucleotide binding by approximately 4-fold and partially inactivated rab5. Mutations in several other conserved residues (K22A, F57Y, and R81A) also resulted in partial loss of rab5 activity. Eight mutations in and around the putative effector domain had little effect on rab5 activity. In light of these data, the structure-function relationship of rab5 is discussed and compared with that of ras, the prototype of small GTPases. PMID- 8227000 TI - Escherichia coli topoisomerase IV. Purification, characterization, subunit structure, and subunit interactions. AB - DNA sequence analysis of Escherichia coli parC and parE, encoding the subunits of topoisomerase IV (Topo IV) (Kato, J.-I., Suzuki, H., and Ikeda, H. (1992) J. Biol. Chem. 267, 25676-25684), showed that ParC was 22 amino acids longer on the N terminus and ParE was 29 amino acids longer on the C terminus than reported previously. E. coli strains bearing bacteriophage T7 RNA polymerase-based expression plasmids carrying both intact and truncated parC and parE were used to overproduce the ParC and ParE proteins. Full-length ParC and ParE were required to reconstitute Topo IV activity, whereas the truncated ParC and ParE were inactive. Topo IV activity was supported only by ATP or dATP. The [ATP]1/2 for DNA relaxation was 0.45 mM, almost 25-fold higher than the [ATP]1/2 for decatenation of kinetoplast DNA. Topo IV activity was inhibited by the quinolone and coumarin antibiotics, although the concentrations required for 50% inhibition of activity were 3-30-fold higher than those required to inhibit DNA gyrase. The norfloxacin-induced DNA cleavage patterns of Topo IV and DNA gyrase were distinct but overlapping. The native forms of ParC and ParE were a dimer and a monomer, respectively; whereas the active form of Topo IV was a heterotetramer, ParC2ParE2. The inactivity of the truncated forms of ParC and ParE could be attributed to their failure to form the heterotetramer. PMID- 8227001 TI - Transcriptional regulation of puc operon expression in Rhodobacter sphaeroides. Involvement of an integration host factor-binding sequence. AB - The putative overlapping consensus sequences (-129 to -105) for binding of fumarate nitrate reductase regulator- and integration host factor (IHF)-like proteins to puc operon upstream DNA of Rhodobacter sphaeroides was protected from DNase I digestion by purified Escherichia coli IHF. The binding of E. coli IHF to the purported IHF-binding site in the puc upstream DNA is highly sequence specific. The recorded binding affinity was significantly lower than that of E. coli IHF to the lambda attP site. Employing site-directed changes in the DNA sequence within the -129 to -105 region, a loss in IHF binding, as monitored through gel retardation analysis, was correlated with alterations in puc operon expression monitored through the use of puc::lacZ transcriptional fusions. These results suggest that the IHF-binding site is involved in repression of puc operon transcription by oxygen as well as modulation of puc operon transcription levels by incident light intensity. Mutations specific to the upstream half of the putative fumarate nitrate reductase regulator-binding site of the puc upstream DNA did not show any physiological effects under the experimental conditions employed. Taken together, these studies reveal that the DNA sequence between -129 to -105 may involve facilitation of the interaction between upstream and downstream cis-acting regulatory sequences involved in puc operon expression. PMID- 8227002 TI - Kallistatin: a novel human serine proteinase inhibitor. Molecular cloning, tissue distribution, and expression in Escherichia coli. AB - We have recently purified a novel human serine proteinase inhibitor (serpin), designated as kallistatin, which binds to tissue kallikrein and inhibits kallikrein's kininogenase and amidolytic activities. In the present studies, we have cloned a full-length cDNA encoding kallistatin from human liver RNA by the polymerase chain reaction. The cDNA is 1284 base pairs in length and encodes 427 amino acid residues, including a 26-residue signal peptide and a 401-residue mature peptide. The translated amino acid sequence of kallistatin matches with the protein sequence and shares 44-46% sequence identity with human alpha 1 antichymotrypsin, protein C inhibitor, corticosteroid-binding globulin, alpha 1 antitrypsin, thyroxin-binding globulin, and rat kallikrein-binding protein. Kallistatin is a new member of the serpin superfamily with a unique reactive site P1-P1' of Phe-Ser. Four potential glycosylation sites are found in the translated amino acid sequence of kallistatin. In a Southern blot analysis following reverse transcription and polymerase chain reaction, kallistatin was found to be expressed in human liver, stomach, pancreas, kidney, aorta, testes, prostate, artery, atrium, ventricle, lung, renal proximal tubular cell, and a colonic carcinoma cell line T84. A genomic Southern blot using the full-length kallistatin cDNA probe revealed simple banding patterns suggesting the gene encoding kallistatin is single-copied. The kallistatin cDNA encoding the mature peptide was expressed in Escherichia coli. The recombinant kallistatin forms an SDS-stable complex with 125I-human tissue kallikrein and has a molecular mass of 40 kDa. The cloning of human kallistatin cDNA established the identity of the novel kallikrein inhibitor and its expression in a functional form in E. coli provides means for studying its structure-function relationship through protein engineering. PMID- 8227003 TI - Regulation of phospholipase A2 activation by phosphorylation in mouse peritoneal macrophages. AB - The regulation of phospholipase A2 (PLA2) activation by phosphorylation, and phosphorylation of an 85-kDa, arachidonoyl-hydrolyzing PLA2 was investigated in mouse peritoneal macrophages. Phorbol 12-myristate 13-acetate (PMA) and okadaic acid, an inhibitor of serine/threonine phosphatases, stimulated arachidonic acid release, supporting a role for phosphorylation events in regulating PLA2 activation. In response to zymosan, PMA, or A23187, arachidonic acid was released at a linear rate up to 30-45 min after stimulation, whereas there was a 30-min lag preceding arachidonic acid release in response to okadaic acid. The 85-kDa PLA2 was phosphorylated on serine in the macrophages, and the level of phosphorylation increased in response to zymosan, PMA, okadaic acid, and, to a lesser extent, A23187. Two-dimensional phosphopeptide mapping revealed multiple phosphopeptides, several of which showed increased phosphorylation in response to zymosan, okadaic acid, and PMA. Zymosan, PMA, A23187, or okadaic acid stimulated time-dependent increases in PLA2 activity in the cytosolic fraction. PLA2 activation was most rapid in response to PMA, whereas activation in response to okadaic acid was delayed similar to the time course of arachidonic acid release. The cytosolic PLA2 had characteristics of the 85-kDa enzyme, including kinetic properties and substrate preference. Phosphatase treatment of the cytosols dephosphorylated the 85-kDa PLA2 and reversed the increase in activity. The results provide evidence that phosphorylation of the 85-kDa PLA2, induced by stimuli that induce arachidonic acid release, is an important mechanism for activation of the enzyme in macrophages. PMID- 8227004 TI - Optimization of an anti-HIV hairpin ribozyme by in vitro selection. AB - We have applied in vitro selection methods to achieve a large increase in the catalytic activity of a hairpin ribozyme targeted against a highly conserved 14 nucleotide sequence within HIV-1 pol RNA. The substrate specificity was changed by mutating 8 bases within the substrate-binding domain of the parental (-)sTRSV ribozyme. The resulting enzyme cleaved the HIV substrate specifically but with a 20-fold reduction in catalytic efficiency (kcat/KM). Following random mutagenesis, ribozymes with increased activity against the target sequence were selected through 10 rounds of in vitro selection. Selective pressure was increased by decreasing MgCl2 and spermidine concentrations, and reducing reaction time. Variant ribozymes with base substitutions A11-->G and U39-->C were selected in the population. These mutations were introduced singly and in combination into the trans-acting anti-HIV ribozyme. Each of the single-base substitutions significantly increased ribozyme activity, while the activity of double mutant was increased to nearly the level of the parental ribozyme. These findings demonstrate that in vitro selection is a powerful and efficient method to optimize ribozymes for the catalytic inactivation of targeted RNA molecules. PMID- 8227005 TI - Specific interaction with rhodopsin is dependent on the gamma subunit type in a G protein. AB - Interaction with a receptor is the first step in the process of signal transduction by heterotrimeric (alpha beta gamma) G proteins. We have examined the role of the G protein gamma subunit in interaction between a receptor (rhodopsin) and a G protein, transducin (Gt). We have obtained recombinant beta gamma complexes containing the same beta subunit but three different gamma subunit types by expressing them in the baculovirus/insect cell system. We show that the different recombinant beta gamma complexes interact equally well with the alpha subunit of transducin (alpha t) but only the gamma subunit specific to rod photoreceptors (gamma 1) is able to support interaction of alpha t with rhodopsin. This indicates a direct role for the G protein gamma subunits, which are a family of proteins with diverse structures, in conferring specificity to receptor-G protein interaction. PMID- 8227006 TI - Identification of bovine glutamate dehydrogenase as an RNA-binding protein. AB - Two RNA binding activities were demonstrated in bovine liver homogenate. One binding protein was isolated by a simple ion exchange and gel filtration protocol and was shown by N-terminal protein sequence analysis to be glutamate dehydrogenase. Using identical RNA substrate and assay conditions, no detectable RNA binding was observed with equimolar amounts of other representative dehydrogenases and proteins. Furthermore, excesses of tRNA, salmon testis DNA, or each of the four homoribopolymers were unable to compete for the RNA-binding site. Total cytosolic RNA, however, successfully prevented binding of radiolabeled RNA substrate. These data are consistent with glutamate dehydrogenase containing a binding site for heteropolymeric RNA with highest affinity for an as yet undefined nucleotide consensus sequence or structure. The potential physiological relevance of these observations is discussed. PMID- 8227007 TI - A small C-terminal region of the Escherichia coli MalT protein contains the DNA binding domain. AB - MalT, the transcriptional activator of the Escherichia coli maltose regulon, is a 901-amino acid-long protein that specifically binds to short, asymmetric nucleotide sequences present in several copies in the promoters of the regulon. We report that the protein structure involved in this specific binding is carried by a small C-terminal part of MalT encompassing the last 95 amino acid residues. This was demonstrated by fusing the last 95 codons of malT to the gene that encodes glutathione S-transferase, purifying the hybrid protein by affinity chromatography, and comparing the DNase I and dimethyl sulfate footprints of the hybrid and of wild-type MalT on different MalT-binding sites. MalT belongs to a large family of prokaryotic transcriptional activators, which share significant homology in their approximately 60-amino acid C-terminal regions. Our result strongly supports the suggestion that the region of homology corresponds to the DNA-binding domain of the proteins in this family. PMID- 8227008 TI - Myristoylation is not required for GTP-dependent binding of ADP-ribosylation factor ARF1 to phospholipids. AB - Membrane binding of ADP-ribosylation factors (ARFs) is GTP-dependent and seems to require amino-terminal myristoylation. Recently it has been proposed that myristoylation is needed not for the activation of ARF by GTP but for its subsequent association to membranes. Here we show that unmyristoylated bovine ARF1, expressed in bacteria, can be efficiently loaded with GTP gamma S (guanosine 5'-O-(thio)triphosphate) at 1 microM free Mg2+, in the presence of phospholipids. Unmyristoylated ARFGTP gamma S cosediments with phospholipid vesicles and totally binds to phospholipid-cholate micelles, as seen by gel filtration chromatography. We therefore propose that, in vivo, myristoylation is required for the interaction of ARFGDP with its membrane-bound exchange factor rather than for the association of ARFGTP with lipid membranes. Phospholipid bound ARFGTP gamma S can also stably interact with and activate the catalytic subunit of cholera toxin, suggesting that ARFGTP provides a membrane anchor for cholera toxin and thereby facilitates its access to membrane-bound substrates. PMID- 8227009 TI - ADP-ribosylation of rho p21 inhibits lysophosphatidic acid-induced protein tyrosine phosphorylation and phosphatidylinositol 3-kinase activation in cultured Swiss 3T3 cells. AB - Botulinum C3 exoenzyme was used to specifically ADP-ribosylate and inactivate rho p21, and the effects of rho p21 inactivation on lysophosphatidic acid (LPA) induced tyrosine phosphorylation were examined in cultured Swiss 3T3 cells. LPA induced a rapid increase in the tyrosine phosphorylation of a number of proteins. Pretreatment of the cells with the C3 exoenzyme caused ADP-ribosylation of rho p21 in the cells and selectively attenuated the phosphorylation of several proteins, including p43 mitogen-activated protein kinase, p125 focal adhesion kinase, and two proteins of 72 and 88 kDa. C3 exoenzyme pretreatment did not block the initial phosphorylation and activation of mitogen-activated protein kinase but suppressed its subsequent rise. In contrast, the enzyme treatment inhibited the induction of phosphorylation of the 72- and 88-kDa proteins and suppressed the basal and LPA-induced tyrosine phosphorylation of p125 focal adhesion kinase. In addition, immunoprecipitation of cell lysates with an antibody directed against the 85-kDa subunit of phosphatidylinositol 3-kinase (PI 3-kinase) co-precipitated a tyrosine-phosphorylated band of 180 kDa. C3 exoenzyme pretreatment suppressed both the phosphorylation of this band and PI 3-kinase activation associated with LPA stimulation. These findings suggest that rho p21 works as a link between the LPA receptor signal and the subsequent tyrosine phosphorylation and PI 3-kinase activation in these cells. PMID- 8227010 TI - Expression cloning of type 2 angiotensin II receptor reveals a unique class of seven-transmembrane receptors. AB - Angiotensin II acts on at least two distinct receptor subtypes (AT1 and AT2). Most known effects of angiotensin II in adult tissues are attributable to the AT1 receptor. The function of AT2 receptor is undefined, but its abundant expressions in fetal tissues, immature brain, skin wound, and atretic ovarian follicles suggest a role in growth and development. Previous studies suggested that AT2 receptor may not be G protein-coupled. Here, from a rat fetus expression library, we cloned a cDNA encoding a unique 363-amino acid protein with pharmacological specificity, tissue distribution, and developmental pattern of the AT2 receptor. It is 34% identical in sequence to the AT1 receptor, sharing a seven transmembrane domain topology. A review of prior data on other receptors suggests that this receptor may belong to a unique class of seven-transmembrane receptors (including somatostatin SSTR1, dopamine D3, and frizzled protein Fz) for which G protein coupling has not been demonstrated. All members of this class exhibit fetal and developmental and/or neuronal-specific expression. A conserved motif in the third intracellular loop, distinguishing this class from "classical" G protein-coupled receptors, may mediate novel intracellular effects. PMID- 8227011 TI - Molecular cloning of a novel angiotensin II receptor isoform involved in phosphotyrosine phosphatase inhibition. AB - There are two major isoforms of the angiotensin II receptor, type 1 (AT1) and type 2 (AT2). AT2 is distinguished from AT1 with respect to its ligand selectivity, its insensitivity to non-hydrolyzable GTP analogues, and its as yet unidentified biological functions. In the present study we have expression-cloned AT2 cDNA from a cDNA library of a rat pheochromocytoma cell line (PC12w). Rat AT2 cDNA encodes a 363-amino acid protein that has seven transmembrane domains. AT1 is the closest in homology to AT2 but with only a 32% identity of amino acid sequence. Stably expressed in COS-7 cells, the receptor showed selective binding to AT2-specific ligands PD123319 and CGP42112A but not to the AT1-specific ligand, losartan. Northern blot analysis revealed that the mRNA of rat AT2 was expressed not only in PC12w cells but also in the adrenal glands and in the inferior olive of the brain, both of which are known to contain AT2 type binding sites. The expressed AT2 receptor mediated angiotensin II-induced inhibition of protein tyrosine phosphatase, an action that was dependent on a pertussis toxin sensitive G-protein-coupled mechanism in COS-7 cells. The AT2-specific ligand CGP42112A was an agonist rather than antagonist in the inhibition of phosphotyrosine phosphatase. AT2 did not cause a decrease in cGMP in PC12w or COS 7 cells expressing AT2 stably. These results indicate that the AT2 receptor is structurally and functionally different from AT1 and suggest novel functional roles of the renin-angiotensin system in cross-talk with phosphotyrosine signaling by modulating protein phosphotyrosine levels. PMID- 8227012 TI - GK* and brain G beta gamma activate muscarinic K+ channel through the same mechanism. AB - A pertussis toxin-sensitive G protein (GK) links muscarinic cholinergic and A1 purinergic receptors with an inwardly rectifying K+ (KACh) channel in cardiac atrial cell membranes. Although the beta gamma subunits of pertussis toxin sensitive G proteins (G beta gamma) have been reported to fully activate this channel, it is not known whether exogenously applied G beta gamma interacts with the KACh channel through the same mechanism as the active subunit of endogenous GK (GK*). We examined in inside-out patches the relationship between the concentration of GTP and KACh channel activity with or without preactivation of the channels by either GTP gamma S (guanosine 5'-3-O-(thio)triphosphate) or G beta gamma purified from bovine brain. In the control, KACh channels were activated by intracellular GTP (with acetylcholine in the pipette) in a positive cooperative manner (Hill coefficient approximately 2.5). As the channels were preactivated by GTP gamma S to progressively higher levels, the GTP channel activity relationship shifted more to the left, but the Hill coefficients of the curves remained the same. The same changes were observed when KACh channels were preactivated with brain G beta gamma. These results indicate that endogenous GK* and exogenous G beta gamma share a common molecular mechanism to activate the KACh channel. PMID- 8227013 TI - Carboxyl terminus is essential for intracellular folding of chloramphenicol acetyltransferase. AB - Chloramphenicol acetyltransferase (CAT, EC 2.3.1.28) is a bacterial chloramphenicol resistance marker that is commonly used as a reporter enzyme in gene expression studies and as a carrier protein for the production of fused peptides. The latter can be done by insertion of target sequences into the native ScaI site near the 3'-end of the Tn9 cat gene. CAT activity in the resulting fusion proteins is retained. We observed that creation of a stop codon at this ScaI, which causes a COOH-terminal 9-amino acid deletion, results in loss of chloramphenicol resistance and total deposition of the mutant protein in inclusion bodies in Escherichia coli. Cytoplasmic solubility and enzyme activity are completely regained by elongation of this mutant with only 2 residues. Apparently, terminal residues of the alpha 5-helix play a crucial role in achieving the native conformation of nascent CAT molecules. Thus, CAT provides an interesting model system for mutational analysis of protein folding in vivo. PMID- 8227014 TI - Phospholipase C activation during elicitation of the oxidative burst in cultured plant cells. AB - Although phospholipase C hydrolysis of polyphosphoinositides constitutes one of the major second messenger pathways in animal cells, its participation in signal transduction in higher plants has not been established. To determine whether activation of phosphatidylinositol-directed phospholipase C might be involved in signaling the elicitor-induced oxidative burst in plants, suspension-cultured soybean cells were treated with two stimulants of the H2O2 burst and examined for polyphosphoinositide turnover. Both polygalacturonic acid elicitor and the G protein activator, mastoparan, promoted a transient increase in inositol 1,4,5 trisphosphate (IP3) content that exceeded basal IP3 levels (0.9 +/- 0.4 pmol of IP3/10(6) cells, n = 28) by 2.6- and 7-fold, respectively. In each case, intracellular IP3 content reached a maximum at 1 min post-stimulation and declined to near basal levels during the subsequent 5-10 min. Neomycin sulfate, an inhibitor of polyphosphoinositide hydrolysis, blocked the IP3 transient, and Mas-17, an inactive analogue of mastoparan, induced no change in IP3. Thin layer chromatography of lipid extracts of the soybean cells corroborated the above results by revealing a rapid decrease in phosphatidyl-inositol monophosphate and phosphatidylinositol 4,5-bisphosphate following polygalacturonic acid elicitor and mastoparan (but not Mas-17) stimulation. Since the rise in IP3 preceded H2O2 production and since neomycin sulfate inhibited the appearance of both, we hypothesize that phospholipase C activation might constitute one pathway by which elicitors trigger the soybean oxidative burst. PMID- 8227015 TI - Sequence of the sodium ion pump methylmalonyl-CoA decarboxylase from Veillonella parvula. AB - The genes encoding methylmalonyl-CoA decarboxylase from Veillonella parvula were cloned on plasmids using oligonucleotides derived from N-terminal amino acid sequences as specific probes. The entire DNA sequence of the methylmalonyl-CoA decarboxylase genes together with upstream and downstream regions was determined. The genes encoding subunits alpha (mmdA), delta (mmdD), epsilon (mmdE), gamma (mmdC), and beta (mmdB) of the decarboxylase were clustered on the chromosome in the given order. The previously unnoted epsilon-chain (M(r) 5,888) was clearly shown to be a subunit of the decarboxylase by correspondence of the N-terminal amino acid sequence with that deduced from the DNA sequence of mmdE. The alpha subunit was 60% identical with the carboxyltransferase domain of rat liver propionyl-CoA carboxylase, the beta-subunit showed 61% sequence identity with the beta-subunit of oxaloacetate decarboxylase from Klebsiella pneumoniae, and the biotin-containing gamma-subunit was 29-39% identical with biotin-domains of other biotin enzymes. The delta-subunit of methylmalonyl-CoA decarboxylase and the gamma-subunit of oxaloacetate decarboxylase did not show significant sequence homology. The gross structure of both proteins, however, was similar, consisting of a hydrophobic membrane anchor near the N terminus, a proline/alanine linker, and a remarkable accumulation of charged amino acids in the C-terminal part. The sequence of the small epsilon-subunit could be aligned to the C-terminal region of the delta-subunit downstream of the proline/alanine linker, where the two subunits were 47% identical. Of considerable interest for the mechanism of Na+ transport are the long stretches of complete sequence identity between the hydrophobic beta-subunits of methylmalonyl-CoA decarboxylase and oxaloacetate decarboxylase and the presence of two conserved aspartic acid residues within putative membrane-spanning helices. PMID- 8227016 TI - Effect of mutations at active site residues on the activity of ornithine decarboxylase and its inhibition by active site-directed irreversible inhibitors. AB - Mouse ornithine decarboxylase (ODC) and mutants changing residues thought to be involved at the active site were expressed in Escherichia coli, purified to homogeneity by affinity chromatography on a pyridoxamine 5'-phosphate-agarose affinity column, and tested for their kinetic properties and their inactivation by enzyme-activated irreversible inhibitors. All of the mutant enzymes were expressed at comparable levels to the wild type protein (2-4% of the total soluble protein), all bound to the affinity column, and there were only small differences in the apparent Km values for L-ornithine providing strong evidence that the mutations did not lead to any gross changes in the protein structure. The mutation K69A led to a change in the spectrum of the enzyme and a 550-fold decrease in the kcat/Km (specificity constant) value. These results are consistent with lysine 69 being the residue that forms a Schiff base with the pyridoxal 5'-phosphate co-factor. Mutation C70S did not greatly affect the activity despite its proximity to this lysine but increased the Km about 2-fold. In contrast, the mutation C360A greatly reduced the specificity constant (by 26 fold) despite a 2-fold decrease in the Km, suggesting that this cysteine residue is critically involved at the active site. Although cysteine 360 is known to be the major site of binding of the inhibitor, alpha-difluoromethylornithine (DFMO), the C360A mutant was still sensitive to inhibition by this drug. However, the kinetics of inactivation were altered, the partition ratio was 10 times greater, and the labeled adduct formed by reaction with [5-14C]DFMO was removed from the protein under some denaturing conditions. This adduct was found to occur at lysine 69. The K69A mutant was also sensitive to DFMO with a lower partition ratio than the wild type enzyme. These results indicate that inactivation of ODC by DFMO can occur via interaction with either of two separate residues that form essential parts of the active site. This renders it unlikely that resistant mutants will arise from changes in the enzyme structure. In contrast to the results with DFMO, the C360A mutant ODC was completely resistant to inactivation by (R,R)-delta-methyl-alpha-acetylenicputrescine and was much less sensitive than the wild type enzyme to alpha-monofluoromethyldehydromethylornithine, showing that the reactive species formed from these inhibitors either cannot be formed by this mutant or are unable to react with lysine 69.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8227017 TI - Cloning of a gene (PSD1) encoding phosphatidylserine decarboxylase from Saccharomyces cerevisiae by complementation of an Escherichia coli mutant. AB - A gene (PSD1) encoding a phosphatidylserine decarboxylase of Saccharomyces cerevisiae was cloned by complementation of a conditional lethal mutation in the homologous gene in Escherichia coli strain EH150. Expression of the cDNA clone in EH150 corrected growth, phospholipid, and phosphatidylserine decarboxylase activity defects. Expression of the genomic clone in wild type yeast resulted in 20-fold amplification of phosphatidylserine decarboxylase activity. A 1500-base pair open reading frame encodes a 56,558-Da protein with a potential mitochondrial targeting sequence. Upstream regulatory elements found in other enzymes of the phospholipid biosynthetic pathway are present in PSD1. The derived amino acid sequence shows 44 and 35% identity with the phosphatidylserine decarboxylases from Chinese hamster ovary cells and E. coli, respectively. Near the carboxyl terminus is an LGST sequence which, in E. coli, is the site of proteolytic cleavage of the proenzyme into the alpha and beta subunits and formation of the pyruvate prosthetic group (Dowhan, W., and Li, Q.-X. (1992) Methods Enzymol. 209, 348-359). Disruption of the PSD1 gene in a haploid strain of yeast resulted in loss of detectable decarboxylase activity but little alteration of the growth properties or phospholipid composition. These results suggest that yeast has a second phosphatidylserine decarboxylation activity. PMID- 8227018 TI - Site-directed mutagenesis of rabbit muscle phosphofructokinase cDNA. Mutations at glutamine 200 affect the allosteric properties of the enzyme. AB - Full-length cDNA for rabbit muscle phosphofructokinase has been cloned and characterized (Li, J., Chen, Z., Lu, L., Byrnes, M., and Chang, S. H. (1990) Biochem. Biophys. Res. Commun. 170, 1056-1060). The 2.8-kilobase cDNA was inserted in the plasmid vector pPL2 and transformed into Escherichia coli cells deficient in endogenous phosphofructokinase activity (DF 1020). The recombinant phosphofructokinase so prepared is nearly identical in kinetic properties and size of subunits to the enzyme isolated from rabbit muscle. On the basis of the sequence homology between the muscle and the bacterial phosphofructokinases and the crystallographic structure of the latter, the glutamine at position 200 of the muscle enzyme is implicated in the allosteric transitions. This residue was replaced by alanine (Q200A), glutamate (Q200E), or arginine (Q200R). The purified enzymes were analyzed for quaternary structure, activity, and allosteric properties. The native and all the altered enzymes are tetramers. At pH 7.0, the wild-type enzyme is sensitive to inhibition by ATP at concentration above 0.6 mM, and its activity responds to fructose 6-phosphate concentration cooperatively at high ATP concentration. In contrast, the mutated enzyme Q200R is virtually insensitive to ATP inhibition up to 7 mM. Thus at high ATP concentration, its activity responds to fructose 6-phosphate concentration is a manner similar to the activated form of the native enzyme. Under the same conditions, mutant Q200E exhibits cooperative behavior only at much higher concentration of fructose 6 phosphate. Mutant Q200A is active at pH 8.0 but inactive at pH 7.0. The native enzyme and all three mutants are activated by inorganic phosphate and fructose 2,6-bisphosphate and inhibited by citrate. PMID- 8227019 TI - Purification and sequence of rat extracellular superoxide dismutase B secreted by C6 glioma. AB - An enzyme which converts radical oxygen, produced by phorbol 12-myristate 13 acetate activated neutrophils, into nonluminescent products is secreted by rat C6 glioma. The enzyme was purified from chemically defined conditioned media and identified as an extracellular superoxide dismutase (EC-SOD). The purified enzyme is distinct from human EC-SOD C (Hjalmarsson, K., Marklund, S. L., Engstrom, A., and Edlund, T. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 6340-6344) by its elution from heparin-Sepharose at 300-400 mM NaCl, its pI of 6.1-7.2, and its native M(r) of 85,000 +/- 20,000. The rat EC-SOD is a dimer with a subunit M(r) of 34,000 36,000 and is extensively modified by post-translational processing. Although rat EC-SOD has a high sequence homology with the catalytic center and the polybasic heparin-binding site near the COOH terminus of human EC-SOD C, its NH2-terminal sequence and the sequences flanking the heparin-binding site differ substantially. The sequence of the isolated rat EC-SOD cDNA fully confirms the data obtained from amino acid sequence analysis. The amino acid sequence of the enzyme and its biochemical properties support its identification as the rat EC SOD B. PMID- 8227020 TI - Probing the reactivity of the GTP- and GDP-bound conformations of elongation factor Tu in complex with the antibiotic GE2270 A. AB - The activity of Escherichia coli elongation factor Tu (EF-Tu) depends on its GTP- and GDP-bound conformations. In this work we have studied the influence of the antibiotic GE2270 A, a new EF-Tu-specific inhibitor of protein biosynthesis, on these two conformations with respect to the interaction with the various ligands and stability. One molecule of GE2270 A bound per EF-Tu is sufficient to block poly(U)-directed poly(Phe) incorporation. This drug binds stably to both EF Tu.GTP and EF-Tu.GDP but only affects the reactivity of the GTP-bound conformation that is no longer able to interact efficiently with aminoacyl-tRNA (aa-tRNA) and ribosomes. Consequently, the protection by EF-Tu.GTP of the aminoacyl-ester bond against nonenzymatic hydrolysis is strongly weakened. The intrinsic EF-Tu GTPase is little affected, but its response to aa-tRNA and ribosomes is impaired. The specificity of GE2270 A to the GTP-bound form of EF-Tu is supported by the course of temperature- and urea-dependent inactivation or tryptic digestion. Furthermore, in its presence elongation factor Ts (EF-Ts) can enhance the dissociation of EF-Tu.GDP but not that of EF-Tu.GTP. Unlike kirromycin, this antibiotic can bind to EF-Tu.EF-Ts without causing dissociation of this complex. Evidence was obtained that the EF-Tu binding site for GE2270 A is also distinct from those for kirromycin, pulvomycin, and aa-tRNA, even though this antibiotic functionally interferes with all three of these ligands. GE2270 A appears to interact with a crucial regulatory region of the EF-Tu molecule that is activated in the GTP-bound state. PMID- 8227021 TI - Biochemical comparisons of the Saccharomyces cerevisiae Bem2 and Bem3 proteins. Delineation of a limit Cdc42 GTPase-activating protein domain. AB - The Bem2 and Bem3 proteins, which appear to play roles in the regulation of bud site formation in Saccharomyces cerevisiae, show striking homology to a number of proteins that compose a family of GTPase-activating proteins (GAPs) for the rho subgroup of ras-related GTP-binding proteins. These members include human platelet GAP for Cdc42Hs (the human homolog of a S. cerevisiae GTP-binding protein that regulates bud site assembly), the break point cluster region protein, the brain protein chimerin, the 85-kDa regulatory subunit (p85) of the phosphatidylinositol 3-kinase, and the ras-GAP-binding protein (p190). A fusion protein composed of the glutathione S-transferase protein and the rho-GAP homology region of Bem3 (designated GST-Bem3) stimulates the GTPase activity of the wild-type Cdc42Hs protein (Cdc42HsGly-12), but has no stimulatory effect on a GTPase-defective mutant (Cdc42HsVal-12), whereas a GST-Bem2 fusion protein does not stimulate the GTPase activity of either form of Cdc42Hs. We have compared the ability of GST-Bem3 to serve as a GAP for Cdc42Hs relative to other members of the rho-GAP subfamily and found the following order of potency: human platelet Cdc42Hs GAP > p190 > Bem3 > break point cluster region protein, whereas p85, like Bem2, shows no GAP activity or any ability to bind to the GTP-bound form of Cdc42Hs. We have taken advantage of the functional specificity exhibited by Bem3 (versus Bem2) in using Bem2/Bem3 chimeras, as well as different deletion mutant versions of the Bem3 protein, to delineate the limits of a functional Cdc42 GAP domain. The results of this study indicate that the carboxyl-terminal approximately 224 amino acids (which contain three regions of homology to the other members of the rho-GAP family) represent a "limit GAP." The first two appear to be important for binding to Cdc42Hs and for partial GAP activity. PMID- 8227022 TI - Ca(2+)-induced folding and aggregation of skeletal muscle sarcoplasmic reticulum calsequestrin. The involvement of the trifluoperazine-binding site. AB - Calsequestrin is an intermediate affinity, high capacity Ca(2+)-binding protein found in the lumen of the sarcoplasmic reticulum of both skeletal and cardiac muscle cells. Previous sequence analysis suggested that calsequestrin may contain a hydrophobic binding site for the drug trifluoperazine, a site shared by the calmodulin family and shown to play a role in calmodulin/calmodulin receptor interaction. Previous studies showed that, upon Ca2+ binding, calsequestrin undergoes a conformational change, burying the trifluoperazine-binding site, folding into a more compact structure that is trypsin-resistant, and increasing the negative ellipticity of the circular dichroism spectrum. In this study, the structural and functional roles of the trifluoperazine-binding site in the Ca(2+) induced conformational change of calsequestrin are further studied using the calmodulin antagonists trifluoperazine and melittin. If trifluoperazine or melittin is added to calsequestrin prior to Ca2+ addition, then Ca(2+)-induced folding is inhibited as determined by the changes in circular dichroism spectra and protein sensitivity to trypsin digestion. If, however, Ca2+ is added prior to trifluoperazine or melittin, calsequestrin remains resistant to trypsin digestion, just as if the calmodulin antagonists are not present, suggesting that the conformational change is not affected. Aggregates of calsequestrin that exhibit high Ca2+ binding capacity have previously been shown to occur at high Ca2+ and calsequestrin concentrations. By preventing a prerequisite folding step, trifluoperazine or melittin also prevents the Ca(2+)-induced aggregation of calsequestrin, thus decreasing the maximal Ca2+ binding by calsequestrin. These data suggest that the trifluoperazine-binding site is critically involved in the Ca(2+)-induced intramolecular folding step required for the intermolecular interactions leading to high capacity Ca(2+)-binding by calsequestrin. PMID- 8227023 TI - Electron transfer process in milk xanthine dehydrogenase as studied by pulse radiolysis. AB - The reduction of milk xanthine dehydrogenase by salicylate anion radical (SL-), nicotinamide adenine dinucleotide radical (NAD.), and 1-methylnicotinamide (NMA) radicals was investigated by the use of pulse radiolysis. Reduction of the dehydrogenase with SL- proceeded via two phases. From the kinetic difference spectra obtained, the faster and slower phases of reduction represent that of one of the iron-sulfur centers and of FAD, respectively. The rate constant of the faster phase increased with the concentration of the enzyme, suggesting that the reduction follows a bimolecular reaction of SL- with the iron-sulfur center. In contrast, the rate constant of the slower phase (510 s-1) was independent of the concentration of the enzyme at pH 7.5. In order to elucidate the contribution of the molybdenum site in the reaction, a similar reaction was performed with enzyme modified with oxipurinol. In the modified enzyme, the slower phase was lost, whereas the faster phase was not affected. These results suggest that the slower phase is due to intramolecular electron transfer from the molybdenum center to FAD. On the other hand, NAD. reacted predominantly with FAD of the dehydrogenase to form the neutral semiquinone of FAD with a second order rate constant of 1.4 x 10(7) M-1 s-1 at pH 7.5, whereas a similar reaction in the oxidase, which was converted from xanthine dehydrogenase by proteolytical cleavage, was not observed. This suggests that NAD. transfers an electron via the binding site for NAD+ on the dehydrogenase. In contrast, NMA radical reduced only an iron-sulfur center of the dehydrogenase with a second order rate constant of 6.5 x 10(7) M-1 s-1 at pH 7.5. PMID- 8227024 TI - Phosphorylation and active ATP hydrolysis are not required for SV40 T antigen hexamer formation. AB - ATP induces structural alterations in SV40 large T antigen and promotes changes in its interaction with the viral replication origin. We have analyzed nucleotide induced changes in T antigen structure in the absence of origin DNA. Most preparations of immunopurified T antigen contain several discrete species ranging in size from monomers through oligomers larger than hexamers. The predominant species consist of monomers and dimers. Incubation of T antigen with ATP or dATP leads to a dramatic and rapid increase in the appearance of T antigen hexamers. Weakly and nonhydrolyzable analogs of ATP are effective as well, indicating that hexamer formation does not require active ATP hydrolysis. After incubation of T antigen with [gamma-35S]ATP, stable association of the labeled nucleotide with all detectable forms occurs. Removal of greater than 80% of the T antigen phosphate residues does not significantly affect the formation of T antigen hexamers, although changes in the distribution and mobility of the other species of T antigen are apparent. Furthermore, T antigen synthesized in and purified from Escherichia coli and, therefore, presumably un- or underphosphorylated, is capable of forming hexamers. Nucleotide-induced T antigen hexamer formation thus appears to require neither protein phosphorylation nor active ATP hydrolysis. PMID- 8227025 TI - Fluid membranes with acidic domains activate DnaA, the initiator protein of replication in Escherichia coli. AB - Acidic phospholipids in a fluid phase dissociate ADP or ATP tightly bound to DnaA protein and, in the presence of ATP and DNA, can restore an inactive ADP form to full activity (Sekimizu, K., and Kornberg, A. (1988) J. Biol. Chem. 263, 7131 7135). Further studies of the interactions between DnaA protein and lipids have used two functional assays: 1) release of ADP or ATP from DnaA and 2) DNA replication upon rejuvenation of an inactive ADP-DnaA protein complex. Among a variety of phospholipids tested were pure synthetic compounds and the mixtures from Escherichia coli auxotrophs (fabA), which are unable to synthesize unsaturated fatty acids and can be supplemented with different acyl derivatives. Fatty acid composition was determined by gas-liquid chromatography and membrane fluidity by fluorescence spectroscopy using 1,6-diphenyl-1,3,5-hexatriene as a probe. Lipid requirements of DnaA protein were shown to be: 1) phospholipids in a fluid phase (i.e. above the transition temperature), 2) a charged polar head group, 3) a lamellar phase (i.e. hexagonal II structures were inactive), and 4) a certain degree of fluidity imparted by the fatty acids esterified to the glycerol backbone. This conclusion was based on the incorporation of: 1) cholesterol, known to increase the packing of lipids, or 2) a branched fatty acyl derivative, which exhibits a fluidizing effect similar to that of a cis double bond. Both agents demonstrated that membrane fluidity is required for DnaA protein function in vitro, consistent with early studies of chromosome initiation in growing cells. PMID- 8227026 TI - cAMP regulates G-protein alpha i-2 subunit gene transcription in polarized LLC PK1 cells by induction of a CCAAT box nuclear binding factor. AB - Heterotrimeric G-proteins function as signal transducers for a variety of hormone coupled enzyme systems in eukaryotic cells. In LLC-PK1 renal cells, vasopressin stimulated adenylylcyclase activity is regulated in part, by the counterbalancing activity of stimulatory G-proteins (Gs) and inhibitory pertussis toxin-sensitive G-proteins (Gi). Two Gi genes encoding the Gi isoforms G alpha i-2 and G alpha i 3 are expressed in LLC-PK1 cells. In polarized cells, these isoforms are topographically segregated to different membranes, which allows for the selective inhibition of adenylylcyclase by G alpha i-2. The genes encoding these isoforms are similarly regulated in these cells during growth and differentiation but differ in response to steroid hormone signals (Holtzman, E.J., Kinane, T.B., West, K., Soper, B.W., Karga, H., Ausiello, D.A., and Ercolani, L. (1993) J. Biol. Chem. 268, 3964-3975). We now demonstrate after stimulating polarized LLC PK1 cells with forskolin, which raises intracellular cAMP levels 50-fold, G alpha i-2 but not G alpha i-3 protein is increased 3-fold at 12 h and remains elevated above control values by 24 h. In cells stably transfected with G alpha i-2 or G alpha i-3 gene 5'-flanking sequences fused to firefly luciferase cDNA reporter gene, forskolin treatment increased G alpha i-2 transcription 3-fold but inhibited G alpha i-3 transcription by 50% at 12 h. In vivo footprinting of forskolin-treated cells was performed to examine the molecular basis for activation of the G alpha i-2 gene. Protected guanosines were identified in a 135 base pair (bp) area previously associated with enhancer activity of this gene in non-polarized cells. This DNA segment did not contain the classical cAMP response element 5'-TGACGTCA-3'. Utilizing the 135-bp DNA segment as a probe in mobility shift assays, which compared nuclear extracts from cells before and after forskolin treatment, an induced nuclear protein complex was identified. Following systematic reduction and mutation of this DNA segment, a "CCAAT" box motif was identified that bound the induced nuclear protein complex during forskolin induced G alpha i-2 gene transcriptional activation. Induction of this nuclear protein complex was prevented in forskolin-treated cells by cycloheximide. To demonstrate functional activity of the CCAAT box motif, cells were transiently transfected with plasmids encoding either the minimal 135-bp segment or a multimerized CCAAT box segment fused to a Rous sarcoma minimal promoter/firefly luciferase reporter gene.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8227027 TI - Directional regulatory activity of cis-acting elements in the bidirectional alpha 1(IV) and alpha 2(IV) collagen gene promoter. AB - The genes for the type IV collagen alpha 1 and alpha 2 chains are regulated by a common alpha 130-base pair (bp) bidirectional promoter. In an attempt to explore the regulation of expression of both genes, we have constructed a minilocus plasmid containing minigenes for the human alpha 1 (IV) and alpha 2(IV) collagen genes located head-to-head like the endogenous genes. The directional regulation of this promoter was studied by introducing site-directed mutations into the promoter of the minigenes. In transient transfections the levels of transcripts of both minigenes were measured, and the influence of mutations on the direction of transcription was studied. The mutational analysis showed that a CCCTCCC motif and a GC box located in the center of the common promoter as well as GC boxes in exon 1 of the alpha 2(IV) gene are potent activating elements for both genes. The promoter of the alpha 1(IV) and alpha 2(IV) collagen genes contains no classical TATA boxes. Even so, A+T-rich sequence motifs, which have no obvious homology to the TATA box located 25-30 bp upstream of exon 1 in each gene, direct the initiation of transcription predominantly about 30 bp downstream of their location. Disruption of the A+T-rich sequence decreased the activity of the promoter considerably, and in addition, the generated classical TATA boxes hindered maintenance of normal bidirectional transcription and changed the ratio of the alpha 1(IV) and alpha 2(IV) transcripts. The CCAAT box in the alpha 2(IV) chain coding strand activated the promoter in the alpha 2(IV) gene direction, but not in the alpha 1(IV) gene direction. PMID- 8227028 TI - Characterization of mutations in the b subunit of F1F0 ATP synthase in Escherichia coli. AB - Site-directed mutagenesis was used to investigate the restrictions on Ala-79 of the b subunit in F1F0 adenosine triphosphate synthase. This amino acid had been previously identified as particularly sensitive to mutation (McCormick, K. A., and Cain, B. D. (1991) J. Bacteriol. 173, 7240-7248). Mutant uncF (b) genes were placed under control of the lac promoter and monitored for F1F0 ATP synthase function in an uncF(b) deletion strain. Three deleterious bAla-79 mutations were moved to the unc operon in the chromosome by homologous recombination. Decreases in enzymatic activity in the uncF (b) mutant strains resulted from reduced amounts of enzyme. With the exception of the bAla-79-->Pro mutation, high expression of mutant uncF (b) genes resulted in increases in F1F0 ATP synthase activity which were sufficient to overcome the defects. In addition to the decrease in the amount of enzyme, the bAla-79-->Lys mutation affected ATP synthesis to a much greater extent than ATP-driven proton translocation. The evidence supports our earlier hypothesis, in which bAla-79 was proposed to play an important, but not essential, structural role in F1F0 ATP synthase assembly or stability. PMID- 8227029 TI - Inhibition of SDC25 C-domain-induced guanine-nucleotide exchange by guanine ring binding domain mutants of v-H-ras. AB - Guanine-nucleotide exchange is the reaction that controls the activation of H-ras p21. This reaction is stimulated by the guanine-nucleotide exchange factor. In this study we show that H-ras p21 harboring guanine ring binding domain (the conserved NKXD motif) mutations, such as N116I or K117E, are potent inhibitors of H-ras p21 guanine-nucleotide exchange reaction promoted by SDC25C (Saccharomyces cerevisiae SDC25 C-domain gene product), a guanine-nucleotide exchange factor. The inhibition is due to the formation of a stable but catalytically inactive complex consisting of the H-ras mutant and SDC25C. By examining the interaction of v-H-ras(N116I) or v-H-ras(K117E) with SDC25C at different concentrations of guanine-nucleotide, we demonstrate that the mechanism of SDC25C-promoted guanine nucleotide exchange proceeds through the following pathway. First, SDC25C binds to H-ras and forms an intermediate H-ras.SDC25C complex; subsequently, an incoming guanine-nucleotide dissociates the complex, releasing SDC25C from H-ras and causes guanine-nucleotide exchange. This mechanism is similar to the one proposed for Escherichia coli elongation factor Ts-catalyzed guanine-nucleotide exchange. PMID- 8227030 TI - How do "Zn2 cys6" proteins distinguish between similar upstream activation sites? Comparison of the DNA-binding specificity of the GAL4 protein in vitro and in vivo. AB - The GAL4 protein of Saccharomyces cerevisiae is the prototype of a family of transcription factors that contain a "Zn2Cys6" coordination complex in the DNA binding domain. GAL4 activates the transcription of genes involved in galactose and melibiose metabolism by binding to sites that contain one or more copies of a sequence 5'-CGGN5TN5CCG-3'. Other Zn2Cys6 proteins in S. cerevisiae also recognize sequences containing two CGG triplets, but with different spacings between them. In this report we investigate the mechanism by which GAL4 distinguishes its bona fide binding site from similar sequences as well as from bulk genomic DNA. In vitro, GAL4 recognizes with moderate to high affinity a variety of sites of the general formula (A/C)GGN10-12CCG. This level of specificity is apparently insufficient for the activator to carry out its biological role. However, many of the sites to which GAL4 binds in vitro do not support GAL4-activated transcription in vivo. In most cases there is not a quantitative correlation between the relative affinity of a site for GAL4 in vitro and the level of GAL4-dependent transcription supported by it in vivo. These data imply that there is some mechanism in vivo by which the intrinsic binding specificity of GAL4 is modified. PMID- 8227031 TI - Isolation and expression of a cDNA encoding a male-specific rat sulfotransferase that catalyzes activation of N-hydroxy-2-acetylaminofluorene. AB - A cDNA (ST1C1 cDNA) encoding a N-hydroxyarylamine sulfotransferase (HAST-I) was isolated from a liver cDNA library of a male adult rat and was expressed in COS-1 cells. ST1C1 cDNA (1363 base pairs) encoded a protein of 304 amino acids with a molecular mass of 35,768 daltons, which shared 50.7 and 46.1% sequence identity with rat aryl (ST1A1 (PST-1)) and estrogen (rOST) sulfotransferases, respectively. N-terminal amino acid sequences of three digested polypeptide fragments of HAST-I were completely identical with two portions of the ST1C1 amino acid sequence. The profile of age- and sex-related expression of ST1C1 mRNA was quite consistent with changes in the sulfating activity of N-hydroxyarylamine and HAST contents in rat livers. ST1C1 expressed in COS-1 cells catalyzed a sulfation of N-hydroxy-2-acetylaminofluorene (N-OH-AAF) at a rate of 4.98 nmol/mg of protein/min and mediated PAPS (3'-phosphoadenosine-5'-phosphosulfate) dependent DNA binding of N-OH-AAF. Although ASTIV was believed to be responsible for the activation of N-OH-AAF, ST1A1 encoding an arylsulfotransferase ASTIV, showed only a marginal activity in a sulfation and covalent binding of N-OH-AAF. These data clearly indicate that ST1C1 cDNA codes a new form of a male-dominant sulfotransferase (HAST) responsible for the bioactivation of N-hydroxyarylamines in rat livers. PMID- 8227032 TI - Interaction of bacterial luciferase with aldehyde substrates and inhibitors. AB - Bacterial luciferase catalyzes the reaction of FMNH2, O2, and an aliphatic aldehyde to yield the carboxylic acid, FMN, water and blue-green light. The kinetics of the bacterial luciferase reaction were measured by stopped-flow spectrophotometry at pH 7 and 25 degrees C for the series of aldehydes from n heptanal to n-undecanal. The rate of formation of the 4a-hydroperoxyflavin intermediate was dependent on the aldehyde concentration when mixtures of enzyme, FMNH2, and aldehyde were rapidly mixed with O2. At saturating aldehyde, the rate of formation of this intermediate was 100-fold slower than in the absence of aldehyde, demonstrating that an enzyme.FMNH2.aldehyde complex can be formed. Numerical simulation of the time courses for these experiments supported the formation of this intermediate and its direct reaction with O2. The kinetics of the light emitting reaction were dependent upon the chain length of the aldehyde substrate. Although the initial light intensity and the light emission decay rate were different for each aldehyde, the quantum yield for the reaction was independent of the aldehyde used. Luciferase was inhibited by high levels of the aldehyde substrate when the enzyme was assayed by mixing FMNH2 with an aerobic mixture of enzyme and aldehyde. The extent of inhibition was dependent on the particular aldehyde used, and the binding affinity of the aldehyde for the free enzyme increased in parallel with the aldehyde chain length. The kinetics of the formation and decay of the various intermediates were also studied in the presence of n-alkyladehyde analogs. These compounds decreased the rate of formation of the 4a-hydroperoxyflavin intermediate in much the same way as the aldehyde substrate, presumably by the formation of the enzyme.FMNH2.analog ternary complex. PMID- 8227033 TI - Behavior of Cys-707 (SH1) in myosin associated with ATP hydrolysis revealed with a fluorescent probe linked directly to the sulfur atom. AB - 4-Fluoro-7-sulfamoylbenzofurazan (ABDF) has a rather small fluorophore that is linked directly to the sulfur atom of thiols without a flexible alkyl chain (Imai, K., and Toyo'oka, T. (1987) Methods Enzymol. 143, 67-75). In the present study I examined the fluorescent and chemical properties of ABDF as an environmentally sensitive probe for Cys-707 (SH1) of myosin subfragment-1 (S-1) to monitor the behavior of SH1 associated with ATP hydrolysis. ABDF was very stable to long irradiation and nonfluorescent before attachment to thiols, permitting the continuous monitoring of the labeling reaction. The fluorophore was useful as an environmentally sensitive probe for thiol groups in proteins. SH1 of S-1 was specifically labeled with ABDF. When ATP, adenyl-5'-yl imidodiphosphate, and ADP were added to the labeled S-1 (ABD-S-1), the fluorescence intensity at 500 nm increased by 110, 66, and 53%, respectively. Binding of actin to ABD-S-1 resulted in a decrease in the fluorescence by 30%. The fluorophore attached to SH1 was found to be located in a more hydrophobic environment in the presence of ATP than in the absence of ligand. KI fluorescence quenching studies suggested that the binding of ATP causes a movement of SH1 toward a more hydrophobic protein interior, whereas it goes back to the opposite direction after ATP hydrolysis. Thus, ABDF is very useful as an environmentally sensitive fluorescent probe for SH1 that monitors ligand-induced changes in the behavior of the thiol. PMID- 8227034 TI - Dissociation of platelet-activating factor production and arachidonate release by the endomembrane Ca(2+)-ATPase inhibitor thapsigargin. Evidence for the involvement of a Ca(2+)-dependent route of priming in the production of lipid mediators by human polymorphonuclear leukocytes. AB - The production of platelet-activating factor (PAF) and the release of [3H]arachidonate were studied in human polymorphonuclear leukocytes (PMN) stimulated with thapsigargin, an inhibitor of endomembrane Ca(2+)-ATPase. Concentrations of thapsigargin as low as 10-25 nM primed PMN for both PAF production and [3H]arachidonate release in response to the chemotactic peptide (fMLP), whereas concentrations in the range 25-200 nM induced a time- and dose dependent production of PAF, which occurred in the absence of both [3H]arachidonate release and [3H]phosphatidylethanol formation. Studies in fura 2/AM-loaded cells showed that concentrations of thapsigargin that elicited PAF production induced a protracted and long lasting elevation of cytosolic free calcium concentration ([Ca2+]i) between 200 and 700 nM. The lower concentrations primed the cells for a late [Ca2+]i elevation in response to fMLP similar to that elicited by cytochalasin B or ionomycin. PAF production showed a good correlation with the increase of [Ca2+]i (r = 0.91) irrespective of the procedure used to grade [Ca2+]i. In contrast, phorbol 12,13-dibutyrate failed to induce both PAF production and elevation of [Ca2+]i, but it was a very effective stimulator of [3H]arachidonate release and [3H]phosphatidylethanol production. These data indicate that PAF production and [3H]arachidonate release in PMN differ in both biochemical pathway and modulatory mechanisms. Whereas PAF production seems extremely sensitive to changes in [Ca2+]i, which seems to exert its modulatory effect at the lyso-PAF:acetyl-CoA acetyltransferase step, [3H]arachidonate release seems tightly modulated by protein kinase C-dependent mechanisms and is coincidental with activation of phospholipase D. PMID- 8227035 TI - Functional roles of valine 37 and glycine 38 in the mobile loop of porcine cytosolic aspartate aminotransferase. AB - The functional roles of Val37 and Gly38 in porcine cytosolic aspartate aminotransferase have been studied in the site-directed mutants V37A, G38A, and G38S where the size and hydrophobic character of these residues has been altered. Previous x-ray studies have shown that Val37 and Gly38, which are part of a flexible loop, interact directly with bound substrate. From x-ray and solution experiments we find that the V37A, G38A, and G38S mutations do not cause significant perturbations to the unliganded enzyme. Replacing Val37 with a less bulky alanine residue does not affect the maximal catalytic rate (kcat), but it does increase significantly the Michaelis constants for substrates in the overall transamination reaction between aspartate and 2-oxoglutarate. On the other hand, replacing Gly38 with alanine or serine results in striking decreases in kcat to 5 and 0.6%, respectively, of the value observed for the wild-type enzyme, as well as in considerable increases in Km values. Consequently, the catalytic competence, kcat/Km, decreases by 3 orders of magnitude for G38A and by 4 orders of magnitude for G38S. Single turnover reactions of G38A and G38S with four individual substrates (aspartate, glutamate, oxalacetate, and 2-oxoglutarate) are characterized by kinetic parameters that are largely consistent with those of the overall reaction. In addition, the mutations at position 38 impair more seriously the catalytic competence of the enzyme toward C5-substrates than toward C4 substrates. We conclude that Gly38 is probably required for proper function of the enzyme because it permits a high level of flexibility for the 36-39 peptide, which in turn allows the essential substrate-induced movement of the small domain. PMID- 8227036 TI - Multiubiquitin chains linked through lysine 48 are abundant in vivo and are competent intermediates in the ubiquitin proteolytic pathway. AB - A prerequisite for the selective degradation of intracellular proteins by the ubiquitin-dependent proteolytic pathway is the attachment of a chain of ubiquitin monomers to the targeted protein. In these multiubiquitin chains, the carboxyl terminal glycine 76 of ubiquitin is linked via an isopeptide bond to the epsilon amino group of lysine 48 in the adjacent ubiquitin. It remains to be determined whether these chains are preassembled and attached en masse to the target, are made by sequential conjugation of ubiquitin monomers to ubiquitins already linked to the protein, or both. Using the 20-kDa ubiquitin-conjugating enzyme TaUBC7 from wheat, we have generated free, glycine 76-->lysine 48-linked multiubiquitin chains (Ubqn), and have individually purified Ubqn species (n < or = 6) by anion exchange, high pressure liquid chromatography. The migration of these chains during SDS-polyacrylamide gel electrophoresis was indistinguishable from those of major ubiquitin immunoreactive proteins in cell lysates from a variety of eukaryotes suggesting that free, multiubiquitin chains are abundant in vivo. One of these chain members (Ubq2) was purified from wheat and was demonstrated via amino acid sequence analysis of tryptic fragments to consist of two ubiquitin monomers joined via a glycine 76-->lysine 48 linkage. We also show in vitro that purified Ubq2 and Ubq4 are competent in activation by ubiquitin-activating enzyme (E1), in transfer to E2s, and in ubiquitin-protein ligase (E3)-independent conjugation to other Ubqn species and to histones H2A/B. These data demonstrate that multiubiquitin chains exist as free, functional structures in vivo and support the possibility that at least a portion of free ubiquitin is preassembled into multiubiquitin chains prior to its attachment to proteolytic substrates. PMID- 8227037 TI - The anti Z-DNA reactivity of Z-DNA forming sequences is affected by platinum antitumor drugs. AB - The effect of binding of platinum antitumor drugs, cis-diamminedichloroplatinum (II) (cis-DDP) and Pt-pentamidine, on the Z-DNA reactivity of potential Z-DNA forming sequences has been studied by enzyme-linked immunosorbent assay. The results indicate that cis-DDP and Pt-pentamidine increase the Z-DNA reactivity of plasmids containing a (dG-dC)16 insert (pUCZ8) and a native Z-DNA forming sequence of Drosophila hydei (pF18). The molar ratio of platinum bound to nucleotides (rb) to produce 50% of Z-DNA reactivity was 0.10 for cis-DDP:pF18, 0.15 for Pt-pentamidine:pF18, and 0.10 for cis-DDP:pUCZ8 and Pt pentamidine:pUCZ8. The efficacy of Pt-pentamidine to provoke Z-DNA reactivity is 2.5-fold higher than that of cis-DDP. While Pt-pentamidine was capable of inducing Z-DNA reactivity in a GC-rich DNA sequence of the hsp 70 protein of Trypanosoma cruzi (p2M4EO3) and in sequences of pUC8, cis-DDP suppresses Z-DNA reactivity. CD spectra of poly(dG-me5dC).poly(dG-me5dC) modified by the drugs suggest that the increase in Z-DNA reactivity observed in plasmids upon drug binding may be due to shifting towards Z-DNA or a Z-DNA like conformation. PMID- 8227038 TI - ATP and A1 adenosine receptor agonists mobilize intracellular calcium and activate K+ and Cl- currents in normal and cystic fibrosis airway epithelial cells. AB - The effect of purinergic compounds on [Ca2+]i and membrane currents of cell lines derived from the airway epithelium of normal and cystic fibrosis individuals has been investigated. 2-Chloroadenosine (2-CADO), as well as other agonists of the A1 adenosine receptors, causes a transient elevation of cytosolic [Ca2+] that is antagonized by the A1 adenosine receptor antagonist 8-cyclopentyl-1,3 dipropylxanthine (DPCPX). ATP is also effective, but at a lower extent. The [Ca2+]i increase induced by 2-CADO and ATP is abolished by preincubation with phorbol 12-myristate 13-acetate and the Ca(2+)-ATPase inhibitor thapsigargin. This latter result suggests that purinergic agonists mobilize Ca2+ from inositol 1,4,5-trisphosphate-sensitive stores. Pertussis toxin completely inhibits the effect of 2-CADO, whereas only it partially affects that of ATP, suggesting the involvement of different types of G proteins. Perforated patch clamp experiments carried out in both current clamp and voltage clamp modes show that 2-CADO and ATP activate K(+)- and Cl(-)-selective membrane currents, with a mechanism inhibited by preincubation with DPCPX and thapsigargin. These data indicate that activation of adenosine A1 receptor, in a similar way to ATP receptor, causes [Ca2+]i increase and ion channels activation through a transduction mechanism that is not impaired in cystic fibrosis airway epithelial cells. PMID- 8227039 TI - Purification and characterization of an enzyme involved in oxidative carbon carbon bond cleavage reactions in the methionine salvage pathway of Klebsiella pneumoniae. AB - The 5-methylthio-D-ribose moiety of 5'-(methylthio)-adenosine is converted to methionine in a wide variety of organisms. 2,3-Diketo-5-methylthio-1 phosphopentane is an advanced intermediate in the methionine recycling pathway present in the Gram-negative bacterium Klebsiella pneumoniae. This unusual metabolite is oxidatively cleaved to yield formate (from C-1), 2-keto-4 methylthiobutyrate (the transamination product of methionine), and 3 methylthiopropionate. To further characterize this oxidative conversion, the desthio analog of the naturally occurring diketone, namely 2,3-diketo-1 phosphohexane I, was synthesized. If the metabolism of I is analogous to that of 2,3-diketo-5-methylthio-1-phosphopentane it should be converted to formate, 2 ketopentanoate, and butyrate. An enzyme (E-1), which mediates the oxidative conversion of I to formate and 2-ketopentanoate, was isolated from extracts of K. pneumoniae. E-1 was purified 100-fold to homogeneity in 10% yield. The native enzyme is a monomeric protein of M(r) 27,000. The activity of E-1 requires magnesium ion as a cofactor. No other prosthetic groups were detected. Incubation of the enzyme with I, under anaerobic conditions, led to the discovery of two intermediates. These species have been identified by 1H and 13C NMR, UV-visible spectroscopy, and model chemistry studies as 2-hydroxy-3-keto-1-phospho-1-hexene II, generated by enolization of I; and 1,2-dihydroxy-3-keto-1-hexene III, generated by enzymatic dephosphorylation of II. Intermediates II and III are released from the active site of the enzyme; III accumulates under anaerobic conditions. Under aerobic conditions, III is non-enzymically oxidized to 2 ketopentanoate, formate, and other products. Compound II was also generated by heating I at pH 7.5 for 7 min. Action of alkaline phosphatase on II produces III. PMID- 8227040 TI - Appendix. Cloning and sequence of the gene encoding enzyme E-1 from the methionine salvage pathway of Klebsiella oxytoca. AB - The methionine salvage pathway converts the methylthioribose moiety of 5' (methylthio)-adenosine to methionine via a series of biochemical steps. One enzyme active in this pathway, a bifunctional enolase-phosphatase called E-1 that promotes oxidative cleavage of the synthetic substrate 2,3-diketo-1-phosphohexane to 2-keto-pentanoate, has been purified from Klebsiella pneumoniae and is characterized in the preceding paper (Myers, R., Wray, J., Fish, S., and Abeles, R. H. (1993) J. Biol. Chem. 268, 24785-24791). We synthesized degenerate oligonucleotides corresponding to portions of the amino terminus of E-1. These oligonucleotides were used as polymerase chain reaction primers on whole genomic DNA from Klebsiella oxytoca. This resulted in an 82-base pair DNA fragment that was used as a hybridization probe to obtain a clone of the E-1 gene from a K. oxytoca gene library. The DNA sequence of the E-1 coding region was determined, and the amino acid sequence of E-1 was deduced. E-1 appears to represent a novel class of enzymes since no homology to known enzymes was found. Cloning the gene from K. oxytoca on a multicopy plasmid leads to overproduction of E-1 enzyme that has properties indistinguishable from those of the enzyme from K. pneumoniae. PMID- 8227041 TI - Reversible association of a 97-kDa protein complex found at the tips of ciliary microtubules with in vitro assembled microtubules. AB - Using a human autoimmune CREST antiserum, we identified a 97-kDa polypeptide in Tetrahymena cilia at the plus ends of ciliary microtubules and mammalian kinetochores (Miller, J. M., Wang, W., Balczon, R., and Dentler, W. L. (1990) J. Cell Biol. 110, 703-714). In this study, we examined the interactions of the ciliary 97-kDa protein with microtubules assembled from purified bovine brain tubulin. The 97-kDa protein binds to microtubules and is released from them with 75 mM MgCl2, the same condition used to release it from ciliary microtubules. The 97-kDa protein-microtubule association can be disrupted by ATP and by adenosine 5'-O-(thio-triphosphate), and this ATP sensitivity requires a soluble factor in the crude 97-kDa protein fraction. Fractionation of the crude 97-kDa protein fractions by gel filtration chromatography or by sucrose gradient centrifugation causes the loss of the microtubule binding activity of the 97-kDa protein. Fractions containing a high molecular weight protein complex (molecular mass, approximately 1500-2000 kDa) from the column or gradient fractions can restore the microtubule binding ability of the 97-kDa protein. These results suggest that the 97-kDa protein is part of a high molecular weight protein complex and that the complex, not the 97-kDa protein alone, is required for the 97-kDa protein microtubule association. The association of the 97-kDa protein with microtubules is sensitive to ATP, which suggests that the 97-kDa protein-microtubule interaction may be regulated by an ATPase(s) or protein kinase(s)/phosphatase(s). PMID- 8227042 TI - The phosphorylation and DNA binding of the DNA-binding domain of the orphan nuclear receptor NGFI-B. AB - NGFI-B is an orphan member of the nuclear receptor superfamily encoded by an immediate-early gene. It is rapidly synthesized and phosphorylated in PC12 cells in response to nerve growth factor (NGF) and other agents and is differentially phosphorylated dependent upon the inducing stimulus. The DNA-binding domain (DBD) of NGFI-B has been expressed in bacteria and purified. The purified protein is phosphorylated by protein kinase A or by extracts from NGF-treated PC12 cells. The phosphorylated residues within the DBD have been identified as Ser-340 and Ser-350. The use of mutants in which either or both of these residues were replaced with alanines revealed that phosphorylation of Ser-350, located within the "A box," a motif necessary for DNA binding by NGFI-B, resulted in a decrease in binding to the NGFI-B response element, while phosphorylation of Ser-340 had little or no effect. These findings demonstrate that phosphorylation of a nuclear receptor DBD results in a change in DNA binding and provides another potential mechanism for regulating NGFI-B activity. PMID- 8227043 TI - The Schizosaccharomyces pombe mating-type gene mat-Mc encodes a sequence-specific DNA-binding high mobility group box protein. AB - The Schizosaccharomyces pombe gene mat-Mc plays a determinative role in the sexual differentiation of the fission yeast. The mat-Mc protein has been suggested to belong to a novel family of so-called high mobility group (HMG) box proteins, characterized by homology to high mobility group-1 and -2 proteins. Several HMG box proteins, including the mammalian sex-determining gene product SRY and the lymphoid transcription factors TCF-1 and LEF-1, have been shown to bind to DNA in a sequence-specific fashion. To analyze possible DNA-binding properties of mat-Mc, we have cloned and expressed its putative HMG box in Escherichia coli. Gel retardation analysis revealed that the mat-Mc HMG box recognizes the AACAAAG heptamer in a sequence-specific fashion. Combined T-->C and A-->I substitutions on both strands of the AACAAAG heptamer, which change the surface of the major groove while leaving the minor groove intact, did not interfere with sequence-specific binding of mat-Mc. Methylation interference analysis confirmed that the mat-Mc HMG box contacts adenine residues in the minor groove. By using a circular permutation assay, the mat-Mc HMG box was observed to bend DNA. These results indicate that mat-Mc is indeed a member of the HMG box family with DNA-binding characteristics assigned earlier to other members of this novel transcription factor family. PMID- 8227044 TI - Alterations in the electron transfer chain in mutant strains of Escherichia coli lacking phosphatidylethanolamine. AB - Inside-out sealed membrane vesicles were prepared from strains of Escherichia coli engineered to be lacking in the major phospholipid of this organism, phosphatidylethanolamine (DeChavigny, A., Heacock, P. N., and Dowhan, W. (1991) J. Biol. Chem. 266, 5323-5332). The energy transducing properties, namely the ability to generate a proton gradient directed inward and to transport electrons to molecular oxygen, were compared to those of membranes isolated from wild type cells containing normal levels of phosphatidylethanolamine. Membranes from both cell types were equal in their ability to oxidize succinate and lactate as well as hydrolyze ATP with the generation of proton gradients of similar magnitude, thus establishing the structural integrity of the membrane barrier and basic functionality of the energy transducing systems in the mutant membranes. However, mutant membranes were reduced by about 80% in their type II NADH dehydrogenase dependent oxidase activity which resulted in a reduced ability to generate a proton gradient using NADH as an energy source. Use of artificial electron acceptors indicated that the level of type II NADH dehydrogenase activity was normal. Whole chain NADH oxidase activity could be restored by addition of short chain analogs of the naturally occurring Q8, even though the level of the Q8 pool in both cell types was the same. These results suggest that the function of Q8 in linking type II NADH dehydrogenase with the terminal oxidase(s) is dependent on the phosphatidylethanolamine content of the surrounding phospholipid matrix. PMID- 8227045 TI - Lipophilic 1,1-bisphosphonates are potent squalene synthase inhibitors and orally active cholesterol lowering agents in vivo. AB - Squalene synthase catalyzes the reductive dimerization of two molecules of farnesyl diphosphate to form squalene at the final branchpoint of the cholesterol biosynthetic pathway. We report herein that isoprenyl 1,1-bisphosphonates and related analogs are potent inhibitors of rat microsomal squalene synthase (I50 = 0.7-32 nM). In addition, members of this family are potent inhibitors of cholesterol biosynthesis in rats on intravenous and oral dosing, as well as cholesterol lowering agents in rats and hamsters. Significant inhibition of cholesterol biosynthesis in rats by lovastatin occurs with a concomitant inhibition of dolichol and coenzyme-Q9 synthesis. In contrast, bisphosphonate 4 has no effect on dolichol and coenzyme-Q9 biosynthesis in rats under conditions where cholesterol biosynthesis is > 90% inhibited. PMID- 8227046 TI - Cytotoxicity of folate-Pseudomonas exotoxin conjugates toward tumor cells. Contribution of translocation domain. AB - Folate-protein conjugates can be nondestructively delivered into a cell's cytoplasm via folate receptor-mediated endocytosis if (i) the target cells express a folate-binding protein, and (ii) if the folate is linked to its attached protein at a site that does not interfere with receptor recognition. Because such conjugates have been observed to remain in endosomal compartments for extended periods following cellular uptake, we decided to evaluate whether release into the cytoplasm might be expedited by inclusion of a translocation domain in the folate-protein construct. To test this possibility, momordin-folate and truncated Pseudomonas exotoxin-folate conjugates (LysPE38 and Cys-PE35), i.e. protein synthesis inhibitors either lacking or containing the desired translocation domain, respectively, were examined for their abilities to block protein synthesis in a variety of cell types. The translocation competent LysPE38 folate construct was found to kill cells six times more rapidly with 10-fold greater potency than the permeation-incompetent mormordin-folate. Further, cells expressing low levels of folate receptors could only be exterminated by the translocation competent Pseudomonas exotoxin-folate conjugates. When the translocation capability of CysPE35-folate was inactivated by modification of Cys287, the construct also lost most of its cytotoxicity. These data suggest that autocatalysis of transport from an internal vesicular compartment into the cytoplasm can greatly augment the cytotoxicity of a protein toxin entering cells via the folate endocytosis pathway. Because the folate ligand can selectively target a protein conjugate to cancer cells in the presence of normal cells, such translocatable toxin-folate constructs warrant further study as a possible treatment for some malignancies. PMID- 8227047 TI - Processivity of mitochondrial DNA polymerase from Drosophila embryos. Effects of reaction conditions and enzyme purity. AB - Mitochondrial DNA polymerase from Drosophila embryos has been characterized with regard to its mechanism of DNA synthesis under the influence of a variety of compounds in moderate salt (120 mM KCl), where the enzyme is most highly active and only moderately processive, and in low salt (30 mM KCl), where it is less active yet most highly processive. Disparate activity and processivity optima were obtained in low salt in the presence of varying pH or MgCl2 and ATP concentrations; in moderate salt, optimal activity and processivity were achieved coincidentally. Whereas no correlation between processivity and activity optima was observed upon addition of polyethylene glycol in either low or moderate salt, the optima were coincident at both salt levels on addition of glycerol. None of the reaction conditions examined allowed DNA polymerase gamma to exhibit maximal activity and processivity concurrently; maximal activity was always achieved in moderate salt and the highest processivity in low salt. However, while limiting the availability of primer termini had no effect on the mechanism of DNA synthesis, we found that the ability of mitochondrial DNA polymerase to copy singly primed M13 DNA was enhanced then diminished during the course of purification, suggesting loss of an accessory factor. PMID- 8227048 TI - Characterization of wheat germ protein synthesis initiation factor eIF-4C and comparison of eIF-4C from wheat germ and rabbit reticulocytes. AB - Eukaryotic protein synthesis initiation factor (eIF)-4C was purified from wheat germ and the molecular weight was calculated to be approximately 19,000 by SDS polyacrylamide gel electrophoresis. A similar molecular weight was determined by gel filtration chromatography indicating that wheat germ eIF-4C is functional as a single polypeptide chain. An efficient in vitro translation system dependent upon the addition of eIF-4C was developed. This system was used to determine the concentrations of eIF-4C required for the half-maximal rate of translation of satellite tobacco necrosis virus RNA, alfalfa mosaic virus RNA 4, and barley alpha-amylase mRNA. No significant differences in the concentrations of eIF-4C required for the translation of these mRNAs were observed, although differences were noted for eIF-4A and eIF-4F. This finding suggests that eIF-4C is not involved in the binding of mRNA to 40 S ribosomal subunits. In heterologous assays, rabbit reticulocyte eIF-4C was as active as wheat germ eIF-4C in the wheat germ eIF-4C-dependent system. In addition, wheat germ eIF-4C substituted for rabbit reticulocyte eIF-4C in in vitro assay systems from rabbit reticulocytes. These results indicate that eIF-4C from wheat and rabbit contain conserved functional domains. PMID- 8227049 TI - The interaction of N-ethyl retinamide with plasma retinol-binding protein (RBP) and the crystal structure of the retinoid-RBP complex at 1.9-A resolution. AB - The three-dimensional structure of bovine plasma retinol-binding protein (RBP) complexed with N-ethyl retinamide has been determined at 1.9-A resolution. The crystals of the complex (space group P2(1)2(1)2(1), a = 46.27, b = 49.11, c = 76.41 A) are isomorphous with those of bovine holo and apoRBP. The final crystallographic R factor is 0.172 for 11,261 observed reflections. The model of the retinoid-RBP complex is nearly identical to that of bovine retinol-RBP complex; the root mean square deviations between the alpha-carbons in the two proteins is 0.15 A. The N-ethyl retinamide molecule fits in the beta-barrel in the same position previously occupied by the vitamin. The ethyl group of the retinoid replaces the retinol hydroxyl group and a water molecule hydrogen bonded to it. This substitution has no consequence on the overall conformation of the protein. The modification of the functional end group of retinol does not lead to an apparent loss of affinity of the retinol analog for apoRBP, as established by means of fluorometric titrations with N-ethyl retinamide. However, the binding of the retinoid to RBP abolishes or greatly reduces the affinity of RBP for transthyretin. This behavior further supports the hypothesis that the area of the entrance of the beta-barrel, which includes the ethyl group of the retinoid bound to RBP, is involved in the interaction with transthyretin. Moreover, it indicates a high degree of complementarity of interacting surfaces between RBP and transthyretin. In fact, the replacement of the retinol hydroxyl group and quite small structural changes in the above region of the RBP molecule drastically affect the protein-protein recognition. PMID- 8227050 TI - A novel receptor tyrosine phosphatase-sigma that is highly expressed in the nervous system. AB - A novel transmembrane receptor protein tyrosine phosphatase-sigma (RPTP-sigma) was cloned from a rat brain stem cDNA library. The extracellular segment of one form of RPTP-sigma contains 824 amino acids and is composed of three immunoglobulin-like and five fibronectin type III (FNIII)-like repeats. The 627 amino acid cytoplasmic region of RPTP-sigma consists of two catalytic domains oriented in tandem. Northern blot analyses indicate that RPTP-sigma is highly expressed in the brain as two major transcripts of 5.7 and 6.9 kilobases (kb). The 5.7-kb transcript is expressed exclusively in the brain while the 6.9-kb species can be detected in the lung and heart, but at significantly lower levels. In situ hybridization studies confirm that RPTP-sigma is localized predominantly in the nervous system and can be detected in the rat as early as embryonic day 12. During embryonic development, RPTP-sigma is expressed extensively in the central and peripheral nervous systems, including the trigeminal and dorsal root ganglia as well as the retina. In adult rat brain, expression is restricted primarily to the olfactory tubercule, cerebellum, and hippocampus. Within the latter structure, RPTP-sigma is present in the pyramidal cell layer and granular layer of the dentate gyrus. Transfection of RPTP-sigma cDNA into human embryonic kidney 293 cells results in the synthesis of a protein with an apparent molecular mass of 200 kDa as detected by immunoprecipitation and immunoblot analyses using polyclonal antibodies against the FNIII-like repeats present in the extracellular domain of RPTP-sigma. The gene for RPTP-sigma has been mapped to distal chromosome 17 in the mouse. PMID- 8227051 TI - Inhibition of HIV-1 gp160-dependent membrane fusion by a furin-directed alpha 1 antitrypsin variant. AB - Furin is a membrane-associated calcium-dependent serine endoprotease that cleaves proproteins on the carboxyl side of the consensus sequence -Arg-X-Lys/Arg-Arg-. Using site-directed mutagenesis, a variant alpha 1-antitrypsin (alpha 1-AT) was constructed which contains in its reactive site -Arg-X-X-Arg-, the minimal sequence required for efficient processing by furin (Molloy, S. S., Bresnahan, P. A., Leppla, S. H., Klimpel, K. R., and Thomas, G. (1992) J. Biol. Chem. 267, 16396-16402). This alpha 1-AT variant, [Arg355 Arg358]alpha 1-AT (alpha 1-PDX), is greater than 3,000-fold more effective than [Arg358]alpha 1-AT (alpha 1-AT Pittsburgh, alpha 1-PIT) at inhibiting furin in vitro (K0.5 = 0.03 microgram/ml). Furthermore, the P4 Arg in alpha 1-PDX greatly attenuates the thrombin inhibitory properties of this serpin (> 300-fold) compared with alpha 1-PIT (which contains a P4 Ala), thus increasing the selectivity of alpha 1-PDX for furin. Expression studies show that alpha 1-PDX, and not alpha 1-PIT, blocks the processing of two furin substrates, pro-beta-nerve growth factor and human immunodeficiency virus (HIV)-1 gp160 in transfected cells. In addition, a syncytium assay shows that alpha 1-PDX blocks the membrane fusogenic properties of HIV-1 gp160. The potential use of alpha 1-PDX in manipulating the activation of proproteins in a tissue- and time-specific manner is discussed. PMID- 8227052 TI - Characterization of the cell adhesion molecule gp24 in Dictyostelium discoideum. Mediation of cell-cell adhesion via a Ca(2+)-dependent mechanism. AB - Dictyostelium discoideum cells express EDTA-sensitive cell-cell adhesion sites soon after the initiation of development and EDTA-resistant adhesion sites later at the aggregation stage. A glycoprotein of M(r) 24,000 (gp24) has been implicated in the mediation of the EDTA-sensitive type of intercellular cohesiveness (Knecht, D. A., Fuller, D. L., and Loomis, W. F. (1987) Dev. Biol. 121, 277-283). In this study, a relatively simple procedure was developed to purify gp24 to homogeneity. A highly specific rabbit antiserum was raised against gp24, and the localization of gp24 at the cell surface was shown by quantitative binding of the anti-gp24 antibodies to intact cells. To demonstrate the cell binding activity of gp24, the binding of solubilized gp24 to intact cells was examined. 125I-Labeled gp24 bound to cells in a dose-dependent and saturable manner, and the binding was displaced specifically by unlabeled gp24. Purified gp24 was capable of inhibiting the reassociation of dispersed cells previously undergoing EDTA-sensitive aggregation. Moreover, precoating cells with anti-gp24 IgG and Fab fragments blocked the binding of 125I-labeled gp24 to cells. Collectively, these in vitro assays provide direct evidence that gp24 is a cell adhesion molecule that most likely functions through a homophilic mode of interaction. The binding of gp24 to cells was sensitive to EGTA, suggesting that the activity of gp24 may involve calcium ions. Binding studies showed that 45Ca2+ could bind to gp24 blotted onto nitrocellulose membrane. In addition, preincubation of the native protein with calcium ions resulted in a shift in its gel mobility. It is therefore likely that gp24 mediates cell-cell interactions via a Ca(2+)-dependent mechanism, rendering gp24 the first cell adhesion molecule in D. discoideum to utilize a Ca(2+)-based adhesion system. PMID- 8227053 TI - Biosynthesis of the prohormone convertase PC2 in Chinese hamster ovary cells and in rat insulinoma cells. AB - The biosynthesis of the prohormone convertase PC2 was studied in Chinese hamster ovary cells stably transfected with PC2 cDNA (CHO/PC2) and in rat insulinoma cells (Rin5f). The major form of PC2 synthesized by CHO/PC2 cells was a 75-kDa protein corresponding to proPC2; this protein was retained intracellularly for 2 4 h following synthesis, suggesting prolonged intracellular residence. In contrast, the major form of PC2 within Rin cells initially exhibited a molecular mass of 72 kDa and was then progressively converted to a 64-kDa species. This 64 kDa species, which required 1-2 h to be released, was the major PC2 form detectable in Rin cell medium. Calcium-dependent benzyloxycarbonyl-Arg-Ser-Lys Arg-aminomethylcoumarin cleaving activity was found in spent Rin cell medium; this activity could be immunoprecipitated with a carboxyl-terminal PC2 antibody, but not with preimmune serum. In neither cell line did intracellular PC2 become endoglycosidase H-resistant over time. PC2 released from Rin cells was also endoglycosidase H-sensitive. Microsequencing and endoglycosidase H results indicate that 75-kDa CHO cell PC2 and 72-kDa Rin cell PC2 both represent proPC2. We speculate that (a) PC2 undergoes unusual glycosylation, which may be related to its slow release from cells, and (b) the 64-kDa molecule detectable in spent Rin cell medium represents the enzymatically active form of PC2. PMID- 8227054 TI - Essentiality of the three carboxyl-terminal amino acids of the plasmid RK2 replication initiation protein TrfA for DNA binding and replication activity in gram-negative bacteria. AB - In a previous study of mutations in trfA, the gene encoding the replication initiation protein of the broad host-range plasmid RK2, a carboxyl-terminal deletion of 3 amino acids of the TrfA protein was found to be completely nonfunctional for RK2 replication in Escherichia coli and other Gram-negative bacteria. In this work site-directed mutagenesis of the trfA gene was carried out to construct TrfA proteins altered in the three carboxyl-terminal positions. Specifically, TrfA proteins with deletions or substitutions of the terminal cysteine, lysine, and arginine (codons 380-382, respectively) were constructed and characterized for their ability to initiate replication from an RK2 origin in vivo in E. coli, Azotobacter vinelandii, Pseudomonas putida, and Agrobacterium tumefaciens and for binding activity to the iterons at the replication origin. Substitutions of the cysteine at position 380 with a glycine or an arginine resulted in a TrfA protein defective in binding to the RK2 origin and, therefore, defective in replication initiation activity in all four Gram-negative bacteria. Substitution of a serine at that position preserved limited function in replication and DNA binding. The lysine at position 381 could be changed to a glutamine without any obvious change in TrfA function. Deletion of the terminal arginine at position 382 did not affect the ability of TrfA to bind to origin iterons but caused a complete loss of replication activity in all four bacteria. Substitution of this terminal arginine with alanine, serine, or glutamic acid also produced replication-defective TrfA protein in all four bacterial hosts while not affecting iteron binding activity. However, substitution of this arginine with a lysine resulted in a loss of replication activity in E. coli and A. vinelandii but had no effect in P. putida and A. tumefaciens. These observations suggest that the terminal arginine plays an essential role in the activity of the TrfA protein, possibly interaction with host proteins, which can be separated from its iteron binding activity. PMID- 8227055 TI - Molecular basis of the alcohol dehydrogenase-negative deer mouse. Evidence for deletion of the gene for class I enzyme and identification of a possible new enzyme class. AB - The molecular basis of the alcohol dehydrogenase (ADH)-negative deer mouse (Peromyscus maniculatus) has been investigated. Several classes of mammalian ADHs have been recognized based upon biochemical and structural properties. ADH cDNA clones identified by hybridization to a mouse class I ADH cDNA clone were obtained from a deer mouse ADH-positive liver cDNA library. This cDNA has been identified as being a class I sequence and represents the deer mouse Adh-1 gene. An additional cDNA sequence identified in both the ADH-positive and -negative deer mouse cDNA libraries was identified by weak cross-hybridization to the mouse cDNA. This cDNA encodes an amino acid sequence representing a new class of mammalian ADH, and the deer mouse gene for this ADH is named Adh-2. ADH-negative deer mice do not produce mRNA, that is detected by the Adh-1 cDNA probe. However, both stocks of deer mice produce high levels of Adh-2 mRNA in liver. Southern analysis using an essentially full-length Adh-1 cDNA probe has shown that the Adh 1 gene is deleted in the ADH-negative mice. Biochemical analysis of enzyme activity suggests at least three ADH polypeptides are expressed in different tissues and have somewhat different substrate specificities, as in the mouse. PMID- 8227056 TI - Effects of integration and replication on transcription of the HIV-1 long terminal repeat. AB - The activity of a promoter is influenced by chromosomal and cell cycle/replication context. We analyzed the influences of integration and replication on transcription of the human immunodeficiency virus (HIV)-1 long terminal repeat (LTR). We found that one requirement for Tat trans-activated expression differed for integrated versus unintegrated HIV-1 LTR. Specifically, the second coding exon of Tat, previously regarded as functionally dispensable, plays a role in the optimal activation of integrated LTRs. We also found that the transcription profile produced from the HIV-1 LTR is influenced by replication. Autonomously replicating vectors that contain the HIV-1 LTR produced patterns of transcription different from counterpart vectors that do not replicate. Both replicating and nonreplicating HIV-1 LTRs remained responsive to Tat. PMID- 8227057 TI - Gene structure of human mitochondrial ATP synthase gamma-subunit. Tissue specificity produced by alternative RNA splicing. AB - We completely sequenced the human gene for ATP synthase gamma-subunit, which was approximately 23 kilobases long and was composed of 10 exons. Exons 1 and 2 encoded the N-terminal presequence required for mitochondrial import, while exons 9 and 10 encoded the C-terminal portions of mature protein. Enzymatic amplification of human heart and liver cDNAs using the polymerase chain reaction revealed two mRNA transcripts that were predicted to encode two 30-kDa isoforms of the gamma-subunit, which differed by the addition of a single amino acid (Asp273) at the C terminus of the liver type isoform. These two mRNA transcripts of the heart (H) type and liver (L) type were generated by alternative splicing of an exon. The same alternative splicing event was observed in bovine tissue. In human tissues, the H type mRNA devoid of exon 9 was expressed specifically in the heart and skeletal muscle, which require rapid energy supply. The L type mRNA was expressed in the brain, liver, kidney etc. Both transcripts were expressed in the skin, intestine, stomach, and aorta. This tissue specificity of transcript heterogeneity suggests the distinct functional or regulatory roles of the gamma subunit isoforms in the catalysis of ATP synthase. This is the first report on tissue-specific isoforms generated by alternative splicing in an energy transducing mitochondrial protein. PMID- 8227058 TI - Post-translational processing of proopiomelanocortin (POMC) in mouse pituitary melanotroph tumors induced by a POMC-simian virus 40 large T antigen transgene. AB - Mice harboring a transgene composed of proopiomelanocortin (POMC) gene promoter sequences (nucleotides -706 to +64) ligated to the simian virus (SV) 40 early gene encoding large T antigen developed large POMC-expressing pituitary tumors. Histologically the tumors arose from the intermediate lobe, contained nuclear SV40 T antigen and POMC peptides, but did not express other pituitary hormones. POMC processing in the pituitary tumors was indistinguishable from normal mouse intermediate lobe melanotrophs and was characterized by high proportions of acetylated and carboxyl-terminal shortened beta-endorphins, and amino-terminal acetylated alpha-melanocyte-stimulating hormone, and virtually no adrenocorticotropic hormone (ACTH)(1-39), beta-lipotropin, or POMC. The tumors contained abundant levels of mRNA for the prohormone convertase PC2 and undetectable levels of PC1. Normal mouse neurointermediate lobe also has a high ratio of PC2/PC1 expression that is distinct from the relative abundance of PC1 in anterior lobe and AtT-20 corticotroph cells. In contrast, extracts from tumors transplanted subcutaneously in nude mice contained predominantly nonacetylated forms of beta-endorphin(1-31) and -(1-27), very little ACTH(1-39), almost no corticotropin-like intermediate peptide or alpha-melanocyte-stimulating hormone, and higher proportions of intact POMC. Surprisingly, despite the less efficient proteolytic cleavage, a transplanted tumor expressed both PC1 and PC2. These studies are the first biochemical documentation of a melanotroph pituitary tumor in a rodent species and provide a new model for the investigation of pituitary oncogenesis and the molecular basis of tissue-specific prohormone post translational processing. PMID- 8227059 TI - Dynamic aspects of DNA/protein interactions in the transcriptional initiation complex and the hormone-responsive domains of the phosphoenolpyruvate carboxykinase promoter in vivo. AB - Transcription initiation of the gene encoding phosphoenolpyruvate carboxykinase (PEPCK) is stimulated by glucocorticoids and glucagon, via cAMP, and dominantly inhibited by insulin in rat liver and H4IIE cells. Lysolecithin-permeabilized H4IIE cells recover completely and continue to multiply, yet are transiently penetrable by macromolecules. These cells, after various hormonal treatments, were utilized for in situ DNase I protection studies of the PEPCK promoter. Nearly all of the sites of protein interaction observed in vitro are protected in vivo as well as several additional sites. The DNase I protection pattern is the same in cells without or with any of the hormone treatments, suggesting that hormonal modulation of transcription does not involve addition or removal of factors from the hormone response elements of the promoter. We focused on the organization and stability of the transcription initiation complex as well as the dynamic nature of distal promoter factors in their interaction with DNA. The transcription initiation complex was detected, and it appears to be co-existent with a short region of naked single-stranded DNA over the TATA box on the template strand, as determined by potassium permanganate reactivity. This complex is quite stable, even under conditions of much reduced RNA synthesis, which suggests that the complex is not broken down and reformed with each round of initiation by RNA polymerase II. Other factors bind to the PEPCK promoter with half-lives ranging from a few minutes to more than 40 min. The cAMP response element apparently involves transcriptional modulation achieved through modification of a bound factor (presumably cAMP response element-binding protein), whereas the glucocorticoid/insulin-responsive region of the promoter functions through factors which are involved in a rapid exchange, suggesting quite different modes of transcriptional regulation. PMID- 8227060 TI - Identification of the vaccinia virus mRNA guanyltransferase active site lysine. AB - The vaccinia virus mRNA capping enzyme is a heterodimeric protein containing subunits of 97 and 33 kDa, the products of genes D1R and D12L, respectively. The enzyme catalyzes the first three reactions in the mRNA cap formation pathway: mRNA triphosphatase, guanyltransferase and (guanine-7-)methyltransferase. The guanyltransferase reaction proceeds by way of a covalent enzyme GMP (E-GMP) intermediate (Shuman, S. and Hurwitz, J. (1981) Proc. Natl. Acad. Sci. U.S.A. 78, 187-191) in which the GMP is linked to the large subunit through a lysine residue (Toyama, R., Mizumoto, K., Nakahara, Y., Tatsuno, T., and Kaziro, Y. (1983) Eur. J. Biochem. 2, 2195-2201; Roth, M. J., and Hurwitz, J. (1984) J. Biol Chem. 259, 13488-13494). In order to identify the map position of the guanyltransferase active site lysine residue, high specific activity [32P]E-GMP was prepared. Digestion of the E-GMP with hydroxylamine at pH 9.5 yielded a 31-kDa radioactive fragment derived from amino acids 1-273. Cleavage of E-GMP with cyanogen bromide produced a radioactive peptide of 14 kDa corresponding to amino acids 242-365. Lysine residues are found at positions 244 and 260. Staphylococcus aureus V8 protease digestion of cyanogen bromide-cleaved E-GMP yields a radioactive product of about 5 kDa in molecular mass corresponding to the peptide generated by cleavage at glutamic acid residues 253 and 297, demonstrating that lysine 260 is the site of linkage of GMP. PMID- 8227061 TI - Impaired biosynthesis of phosphatidylcholine causes a decrease in the number of very low density lipoprotein particles in the Golgi but not in the endoplasmic reticulum of rat liver. AB - We have investigated the mechanism by which inhibition of phosphatidylcholine biosynthesis in rat hepatocytes by choline deprivation causes a reduction in the secretion of very low density lipoprotein (VLDL) (Yao, Z., and Vance, D. E. (1988) J. Biol. Chem. 263, 2998-3004). Rats ingested a choline-deficient or control diet for 3 days, and subcellular fractions of liver were prepared. No change in the amount of apolipoprotein B in the lumina of the endoplasmic reticulum was observed, but there was a 40-50% decrease of apolipoprotein B in the lumina of the Golgi from choline-deficient compared with control rats. Incubation of microsomes, derived from choline-deficient and -supplemented hepatocytes, with trypsin showed similar degradation of apolipoprotein B, indicating similar quantities of this protein are present on the surface and within the lumina. The VLDL particles in the Golgi of liver cells and in plasma, on average, were larger in samples derived from choline-deficient compared with choline-supplemented animals. Incubation of plasma VLDL with proteases demonstrated that the apolipoprotein B of plasma VLDL particles from choline deficient animals had a different susceptibility to digestion than did VLDL from choline-supplemented animals. These data indicate that the number of VLDL particles assembled in the endoplasmic reticulum of liver is similar in choline deficient and -supplemented rats, but the number of particles is decreased in the Golgi from choline-deficient animals. PMID- 8227062 TI - An activation-associated ganglioside in rat thymocytes. AB - During the activation of rat thymocytes elicited by stimulation with 12-O tetradecanoylphorbol-13-acetate (TPA) and calcium ionophore A23187, the total amount of gangliosides per cell was demonstrated to increase, reaching a maximum level several times higher than that of resting cells at 48 h after activation when measured at 24-h intervals. The amount of the overwhelmingly predominant ganglioside in resting thymocytes, GD1c (NeuGc,NeuGc) (Nohara, K., Suzuki, M., Inagaki, F., and Kaya, K. (1991) J. Biochem. (Tokyo) 110, 274-278), was found to increase further as a result of activation. Furthermore, another ganglioside, which was barely recognizable in resting thymocytes, was found to increase in activated and proliferating thymocytes to a level similar to that of GD1c. This activation-associated ganglioside was isolated and its structure examined. On the basis of the results of compositional analysis, methylation analysis, sialidase hydrolysis followed by detection with cholera toxin B subunit on TLC, and proton NMR spectroscopy, this ganglioside was clarified to be a rare species of GD1b containing two N-glycolylneuraminic acid residues. On the other hand, when the thymocytes were activated using concanavalin A (ConA) as a stimulant, the amount of gangliosides per cell was increased more strikingly than that in thymocytes activated with TPA and A23187. In the ConA-activated thymocytes, many other gangliosides, in addition to GD1c and GD1b-(NeuGc,NeuGc), were demonstrated to appear in large amounts. The cause of this difference in gangliosides between thymocytes activated with a combination of TPA and A23187 and those activated with ConA is also discussed. PMID- 8227063 TI - Palmitoylation is required for signaling functions and membrane attachment of Gq alpha and Gs alpha. AB - We have identified the palmitoylated cysteine residues of alpha q and alpha s, alpha subunits of two heterotrimeric G proteins. Mutational substitutions of serines for cysteines 9 and 10 in alpha q and cysteine 3 in alpha s profoundly alter behavior of the subunits expressed in HEK293 cells. Neither mutant alpha subunit incorporates palmitate; both mutant proteins are found in the soluble rather than the particulate fraction; mutant alpha q or alpha s cannot couple a co-expressed receptor to stimulation of phospholipase C or adenylylcyclase, respectively; cysteine substitution prevents a mutationally activated alpha q (R183C) from stimulating phospholipase C directly, and reduces but does not abolish the ability of a similarly activated alpha s (R201C) to stimulate cAMP synthesis. Substitution of a myristoylation sequence for the palmitoylation sites leads to labeling of alpha q and alpha s by myristate, rather than by palmitate. Myristoylation restores the abilities of both nonpalmitoylated alpha q and alpha s to attach to membranes and, in the case of alpha q, restores its ability to stimulate phospholipase C, whether triggered by the R183C mutation or by receptor activation. These findings identify palmitoylation as a critical determinant of membrane attachment for alpha q and alpha s and show that this modification is required for normal signaling by these proteins. PMID- 8227064 TI - Binding of collagen XIV with the dermatan sulfate side chain of decorin. AB - As an approach to elucidate the role of collagen XIV, which is still unclear, molecules exhibiting affinity for this collagen have been sought in connective tissue. Extracts from fetal bovine tendon were resolved by gel electrophoresis and electrophoretically transferred to nitrocellulose. The blot was overlaid with native collagen XIV and the collagen XIV-binding molecules revealed by immunodecoration with a monoclonal antitype XIV collagen antibody. This experimental approach allowed us to reveal in tendon extracts a diffuse band, with an apparent molecular mass of approximately 100 kDa, that binds collagen XIV. This molecule was also found associated with the fractions containing partially purified type XIV collagen. This 100-kDa molecule was sensitive to chondroitinase ABC and, after chondroitinase digestion, yielded a core protein of about 48 kDa. N-terminal sequence analysis of the proteoglycan after blotting allowed us to identify it as decorin. By solid phase assays we have studied this newly described association between decorin and type XIV collagen and shown that it is a saturable process. In addition, preliminary determination of the domains of the two molecules involved in the association has been performed. The possible role of these interactions is discussed. PMID- 8227065 TI - Role of glucokinase and glucose-6-phosphatase in the acute and chronic regulation of hepatic glucose fluxes by insulin. AB - Increased hepatic glucose production (HGP) is the major cause of fasting hyperglycemia in all forms of diabetes. Glucokinase (GK) and glucose-6 phosphatase (Glc-6-Pase) are the proximal and the distal enzymatic steps, respectively, in the regulation of HGP. We examined the impact of changes in GK and Glc-6-Pase activities on in vivo hepatic glucose fluxes in diabetic (D) and control (C) rats. In particular, the acute regulation by insulin was investigated using the euglycemic hyperinsulinemic clamp technique in conscious rats. In experimental diabetes (6 weeks): (a) GK mRNA was decreased by approximately 40%; (b) the Vmax of GK was markedly decreased (approximately 4 versus 9 mumol/g wet weight/min) and that of Glc-6-Pase was 2-fold increased (approximately 30 versus 15 mumol/g wet weight/min, D versus C), while (c) the Km of GK (approximately 10 mM) and Glc-6-Pase (approximately 1.5 mM) were unchanged. HGP was increased by 65% in diabetes and correlated highly with the ratio of Glc-6-Pase/GK (r = 0.81, p < 0.01). Following acute hyperinsulinemia (2 h): (a) GK mRNA increased by approximately 2-fold in both C and D; (b) GK Vmax did not change in C, but doubled to near-normal in D; (c) Glc-6-Pase Vmax decreased by 23% in C and by 34% in D; (d) the Km of GK decreased by approximately 40% (p < 0.01) in C. Acute hyperinsulinemia almost completely inhibited HGP in both C and D, and no correlation was demonstrated between HGP and the ratio of Glc-6-Pase/GK in these groups. Our data suggest that GK and Glc-6-Pase are important determinants of fasting HGP in diabetes. However, acute changes in Glc-6-Pase and GK activities can account for only a small portion of the in vivo inhibition of hepatic glucose flux by insulin, suggesting additional mechanisms for the short-term regulation of HGP. PMID- 8227066 TI - Enhanced activation of the human histone H2B promoter by an Oct-1 variant generated by alternative splicing. AB - POU homeodomain proteins are important regulators of ubiquitous as well as tissue specific transcription. These factors include the broadly expressed octamer motif binding protein Oct-1 and the related cell-specifically expressed protein Oct-2. These two proteins differ in the types of octamer motif-containing promoters they preferentially activate; Oct-1 can activate RNA polymerase II transcription from a small nuclear RNA promoter better than Oct-2, which can better activate an mRNA type promoter. We describe a variant Oct-1-encoding cDNA resulting from two separate alternate splices of the human oct-1 primary transcript; these alternate splices were present in all cell lines tested. This cDNA encodes an amino terminally and carboxyl-terminally truncated form of Oct-1, called Oct-1B, which retains the DNA-binding POU domain and acquires a unique 12-amino acid carboxyl terminal extension. In a transient expression assay, Oct-1B displayed an enhanced ability compared to the larger form of Oct-1 (called Oct-1A in this report) to activate the human histone H2B promoter, an mRNA-type promoter where a natural octamer motif is involved in cell cycle dependent transcription. Thus, the ability of Oct-1 related proteins to activate a natural regulatory target can be influenced by alternative splicing. PMID- 8227067 TI - RNA polymerases IIA and IIO have distinct roles during transcription from the TATA-less murine dihydrofolate reductase promoter. AB - The largest subunit of RNA polymerase II (RNAP II) contains a remarkable region of tandem heptapeptide repeats of the consensus sequence Tyr-Ser-Pro-Thr-Ser-Pro Ser at its carboxyl terminus. This COOH-terminal domain (CTD) is unphosphorylated in RNAP IIA, extensively phosphorylated in RNAP IIO, and absent in RNAP IIB. The reversible phosphorylation of the CTD has been proposed to be integral to each cycle of transcription from the adenovirus-2 major late promoter. The adenovirus 2 major late promoter, however, may not be a good paradigm for the study of CTD function because in vitro transcription from this promoter is not dependent on the CTD. Previous studies suggest that transcription from the murine dihydrofolate reductase (DHFR) promoter requires the CTD. In an effort to investigate the role of the CTD and its phosphorylation, a RNAP II-dependent reconstituted transcription system specific for the DHFR promoter was established. In this reconstituted system, RNAP IIA, but not RNAP IIB, can transcribe from the DHFR promoter. Furthermore, RNAP IIB does not compete with RNAP IIA for preinitiation complex assembly. These results suggest that the CTD plays a critical role in the recruitment of RNAP II to the DHFR promoter. The analysis of preinitiation complexes assembled on the DHFR promoter indicates that RNAP IIA readily assembles into functional preinitiation complexes in contrast to the inefficient assembly of RNAP IIO. However, transcript elongation is catalyzed by RNAP IIO as demonstrated by the photoactivated cross-linking of nascent DHFR transcripts to subunit IIo. These results indicate that transcription from the DHFR promoter involves the reversible phosphorylation of the CTD and support the idea that RNAPs IIA and IIO have essential but distinct functions. PMID- 8227068 TI - Cloning of a membrane-associated protein which modifies activity and properties of the Na(+)-D-glucose cotransporter. AB - An expression library from porcine kidney cortex was screened with a monoclonal antibody (R4A6) which stimulates high-affinity phlorizin binding in kidney and intestine but does not react with the membrane protein (SGLT1) which mediates Na(+)-coupled transport of D-glucose (Hediger, M.A., Coady, M.J., Ikeda, T.S., and Wright, E.M. (1987) Nature 330, 379-381). A cDNA (RS1) was obtained which codes for a hydrophilic M(r) 66,832 polypeptide and contains a predicted hydrophobic alpha-helix at the COOH terminus. After expression in Xenopus oocytes RS1 protein was found associated with the plasma membrane. RS1-homologous mRNAs were detected in renal outer cortex and outer medulla, small intestine, liver, and LLCPK1 cells, but not in skeletal muscle, heart muscle, Madin-Darby canine kidney (MDCK) cells, renal inner medulla, and Xenopus oocytes. After nondenaturing gel electrophoresis of renal brush-border membranes comigration of RS1- and SGLT1-homologous proteins as a high molecular weight complex was demonstrated. RS1 altered the expression of Na(+)-glucose cotransport by SGLT1 in Xenopus oocytes. There was no effect on the expression of the nonhomologous transporters for Na(+)-gamma-aminobutyric acid cotransport and for Na(+) independent glucose transport. However, RS1 also changed the expression of the SGLT1-homologous Na(+)-myo-inositol cotransporter from MDCK cells. The Vmax of methyl-alpha-D-glucopyranoside (AMG) transport expressed after injection of a small amount of SGLT1-cRNA was increased 40-fold when a stoichiometric amount of RS1-cRNA was coinjected. In addition the voltage and glucose dependence of expressed AMG uptake and the concentration dependence of transport inhibition by phlorizin were changed when stoichiometric amounts of RS1-cRNA were coinjected with SGLT1-cRNA. Thus with SGLT1 one apparent transport site (K0.5 about 100 microM) and one apparent phlorizin inhibition site (Ki about 5 microM) was observed whereas with SGLT1 plus RS1 two apparent transport sites (K0.5(1) about 20 microM, K0.5(2) about 1 mM) and two apparent phlorizin inhibition sites (Ki(1) about 0.3 microM, Ki(2) about 30 microM) were found as has been described in brush-border membrane vesicles of kidney and intestine (see e.g. Koepsell, H., Fritzsch, G., Korn, K., and Madrala, A. (1990) J. Membr. Biol. 114, 113-132). The data suggest that the Na(+)-D-glucose cotransporter and possibly also other SGLT1 type Na(+)-cotransporters contain RS1-type regulatory subunits. PMID- 8227069 TI - Isolation of a protein activator of the clathrin-coated vesicle proton pump. AB - An activator of the clathrin-coated vesicle proton translocating ATPase has been purified 1600-fold from bovine brain. The activator, which requires detergent (polyoxyethylene 9-lauryl ether) for release from clathrin-coated vesicles, is heat-stable, trypsin-sensitive, and has an apparent molecular mass of about 6 kDa as determined by high performance liquid chromatography. The activator stimulates the purified H(+)-ATPase of coated vesicles over 50-fold under acidic conditions. Similarly, the activator stimulates proton pumping catalyzed by the reconstituted proton pump. Importantly, this stimulation of proton pumping is observed only when the activator is reconstituted into the interior of the proteoliposomes. Moreover, the activator protein is demonstrated to protect, and co-sediment with, purified proton pump during glycerol gradient centrifugation performed in the presence of ATP. These observations support the notion that this activator serves to determine the pH set point of acidic endomembranes through interactions with the transmembranous sectors of the proton pump. PMID- 8227070 TI - Expression of the complement classical pathway by human glioma in culture. A model for complement expression by nerve cells. AB - In this paper we demonstrate the synthesis of the components of the classical complement pathway, namely C1q, C1r, C1s, C1-Inh, C2, C4, and C5, by human glioma cell lines (U118MG, T193, and T98G). All these components were structurally, antigenically, and functionally similar to their serum counterparts as determined by biosynthetic labeling experiments, Western blot analysis, and hemolytic assays. Northern blot analysis of mRNA demonstrated that, for each of these components, their specific mRNA had the same size as the equivalent mRNA from hepatic tissue. We could not detect the synthesis of C4bp by these cell lines, and the secretion of C1q was only detected after stimulation by interferon-gamma. All these syntheses were up-regulated by interferon-gamma and tumor necrosis factor. Interleukin-1 beta only increased C2 expression and reproducibly down regulated C5 secretion when used at high doses. Glioma cell lines appear to be an efficient and convenient model for the analysis of complement expression in human astrocytic cells. PMID- 8227071 TI - Purification and characterization of a mitogen-activated protein kinase tyrosine phosphatase from Xenopus eggs. AB - The mitogen-activated protein (MAP) kinases are serine-threonine protein kinases that are activated by tyrosine and threonine phosphorylation by the dual specificity protein kinase MEK (MAP kinase/ERK kinase). The present report describes the purification to near homogeneity and characterization of a protein tyrosine phosphatase from Xenopus laevis eggs that dephosphorylates MAP kinase phosphorylated by MEK. Bacterially expressed Xenopus MAP kinase phosphorylated by purified Xenopus MEK was used as substrate throughout the purification. The purification procedure included anion-exchange, cation-exchange, gel filtration, heparin-Sepharose, and chromatography on a column of thiophosphorylated MAP kinase-Sepharose, resulting in a > 3000-fold purification. Upon analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a protein of 47 kDa correlated with activity. The phosphatase showed absolute specificity toward phosphotyrosine and no activity toward phosphothreonyl-phosphoseryl residues of MAP kinase. The pH optimum of the enzyme was 7.0 with a Km of 9.0 microM for phosphorylated MAP kinase. The phosphatase was inhibited by ammonium molybdate (IC50, 2 microM), vanadate (IC50, 250 microM), millimolar concentrations of MnCl2, ZnCl2 and p-nitrophenylphosphate but not by okadaic acid or microcystin. This tyrosine phosphatase may be involved in deactivating MAP kinase in vivo. PMID- 8227072 TI - Characterization of the nucleational core complex responsible for mineral induction by growth plate cartilage matrix vesicles. AB - The factors that drive mineralization of matrix vesicles (MV) have proven difficult to elucidate; in the present studies, various detergent, chemical, and enzyme treatments were used to reveal the nature of the nucleational core. Incubation with detergents that permeabilized the membrane enhanced calcification of treated MV incubated in synthetic cartilage lymph. While detergents removed most of the membrane lipid, they left significant amounts of the MV annexins and nearly all of the Ca2+, Pi, and Zn2+. Extraction with 1 M NaCl removed much of the Ca2+ and Pi present in MV, markedly reducing Ca2+ accumulation; these effects could be prevented by low levels of Ca2+ and Pi in the NaCl extractant. Treatment with chymotrypsin appeared to damage proteins required for MV mineralization; further treatment with detergents to bypass the membrane reactivated MV mineralization. Treatment of MV with pH 6 citrate removed Ca2+ and Pi, destroying their ability to mineralize; subsequent treatment with detergents did not reactivate these MV. Incubation of the detergent-resistant core with o phenanthroline complexed Zn2+ and stimulated mineralization; addition of Zn2+ to synthetic cartilage lymph blocked the ability of the core to mineralize. These studies show that once the nucleational core complex is formed, the membrane enclosed domain is no longer essential for MV calcification. Our findings indicate that the MV core contains two main components as follows: a smaller membrane-associated complex of Ca2+, Pi, phosphatidylserine, and the annexins that nucleates crystalline mineral formation, and a larger pool of Ca2+ and Pi bound to lumenal proteins. These proteins appear to bind large amounts of mineral ions, stabilize the nucleational complex, and aid its transformation to the first crystalline phase. Once nucleated, the crystalline phase appears to feed on protein-bound mineral ions until external ions enter through the MV ion channels. Zn2+ appears to regulate gating of the ion channels and conversion of the nucleational complex to the crystalline state. PMID- 8227073 TI - Angiotensin II stimulates the synthesis of angiotensinogen in hepatocytes by inhibiting adenylylcyclase activity and stabilizing angiotensinogen mRNA. AB - Angiotensin II stimulates the hepatic synthesis and secretion of angiotensinogen, the substrate of renin. In the present study performed on freshly isolated rat hepatocytes we demonstrate that this effect of angiotensin II is mainly related to a transient inhibition of adenylylcyclase. Agents known to decrease intracellular cAMP (angiotensin II, vasopressin, guanfacine) or the cAMP antagonist Rp-adenosine-3',5'-cyclic phosphothioate stimulated, whereas cAMP stimulating agents (isoproterenol, forskolin, glucagon) or the cAMP-agonist Sp adenosine-3',5'-cyclic phosphothioate inhibited angiotensinogen synthesis. In contrast, all agents known to affect intracellular concentrations of calcium, as confirmed in Fura-2-loaded hepatocytes (Bay K 8644, calcimycin, calmidazolium, ionomycin, or methoxamine) failed to influence the synthesis of angiotensinogen. The inhibitory effect of angiotensin II as well as the stimulatory effect of glucagon on cAMP were inversely related to angiotensinogen mRNA and angiotensinogen secretion over a wide concentration range of both peptides. Both the angiotensin II-dependent inhibition of cAMP and the angiotensin II-induced increase in angiotensinogen mRNA were abolished by a pertussis toxin pretreatment. In hepatocyte membranes, pertussis toxin ADP-ribosylated a single protein (approximately 41 kDa) probably representing the alpha-subunit of the Gi protein, coupling inhibitory receptors to adenylylcyclase. We further show that the increase of angiotensinogen mRNA and secretion mainly represents the result of mRNA stabilization, since in a nuclear run-on assay, angiotensin II pretreatment of hepatocytes does not significantly alter the rate of [32P]UTP incorporation into angiotensinogen mRNA, whereas angiotensin II prolonged the half-life of angiotensinogen mRNA in transcription-arrested as well as in [3H]uridine pulse-labeled hepatocytes about 2.5-fold from 80 to 190 min. It is concluded that angiotensin II induces an increase in angiotensinogen synthesis in hepatocytes by stabilizing of angiotensinogen mRNA and that this effect is mediated through inhibition of adenylylcyclase. PMID- 8227074 TI - The agrin receptor. Localization in the postsynaptic membrane, interaction with agrin, and relationship to the acetylcholine receptor. AB - Agrin is a component of the synaptic basal lamina that induces the aggregation of acetylcholine receptors (AChRs) and other elements of the postsynaptic membrane. We have determined the localization, binding characteristics, and biochemical profile of the agrin receptor in Torpedo electric organ membranes and defined domains of agrin that bind this receptor. Postsynaptic membranes from Torpedo electric organ bind agrin as judged by depletion of AChR clustering activity from solution. A ligand-based radioimmunoassay shows that agrin binding to postsynaptic membranes is saturable and calcium-dependent. Half-maximal binding is observed at agrin concentrations < or = 10(-10) M. Identification of the bound agrin polypeptides shows that at least one membrane binding domain of agrin is located in a 70-kDa proteolytic fragment. Immunofluorescent visualization and radioimmunoassay of agrin binding demonstrates that the agrin receptor is selectively concentrated in postsynaptic membranes, with little binding detected on nonsynaptic or liver membranes. Agrin binding is greatly reduced if the membranes are pretreated with trypsin, but is unaffected by phosphatidylinositol specific phospholipase C. Membranes stripped of peripheral proteins by alkaline treatment retain full ligand binding capacity. alpha-Bungarotoxin affinity columns bind AChRs but not agrin receptors. The ratio of agrin receptors to AChRs in postsynaptic membranes is approximately 1:200. We conclude that the agrin receptor is an integral membrane glycoprotein that is selectively concentrated in postsynaptic membranes, but that is not tightly complexed with the AChR. The results also indicate that the biological activity of agrin is mediated through intracellular signal transduction events triggered by ligand binding to the agrin receptor. PMID- 8227075 TI - Identification of a secondary Fc gamma RI binding site within a genetically engineered human IgG antibody. AB - Although human IgG2 is not cytophilic, we have shown previously that an IgG2 antibody expressing the sequence PLLGG (underline = substitution) spanning CH2 domain residues 233-237 (Eu numbering) displayed IgG1-like Fc gamma RI binding activity. In contrast, IgG1 PLLGG exhibited 3-fold less affinity, whereas IgG2 ELLGG was 3-fold more active than native IgG1. These results suggested that additional site(s) conferred enhanced binding properties to the engineered, cytophilic IgG2 variant. These sites were shown to reside in the IgG2 CH2 domain, since the IgG1 CH2 module did not have enhanced activity in a panel of hybrid IgG1/IgG2 antibodies. To map these sites further, human IgG1 and IgG2 constant region gene segments were modified to allow reciprocal COOH-terminal half segment exchanges of CH2 exons. These were cloned into a pSV2neo expression vector bearing a rearranged MOPC 315 heavy chain variable region gene and transfected into a MOPC 315 heavy chain deletion mutant. The dinitrophenol affinity-purified IgGs were radiolabeled and assessed for Fc gamma RI binding activity in direct binding assays using U937 cells. The COOH terminus of the IgG2 CH2 domain was found to contain accessory site(s) since it enhanced the binding properties of both IgG1 PLLGG and native IgG1. In contrast, grafting of the COOH terminus of the IgG1 CH2 domain onto IgG2 PLLGG and IgG2 ELLGG diminished their cytophilic activity. The amino acid responsible for the enhancing properties of the COOH terminus of the IgG2 CH2 domain was shown to be threonine 339, since IgG1 PLLGG/Thr339 displayed increased Fc gamma RI binding affinity. Kinetics studies revealed that this is accomplished through an increase in the forward rate constant of the IgG-Fc gamma RI interaction. PMID- 8227076 TI - 1,25-Dihydroxyvitamin D3 activates Raf kinase and Raf perinuclear translocation via a protein kinase C-dependent pathway. AB - 1,25-Dihydroxyvitamin D3's (D3) potential mitogenic mechanism of action was pursued in cultured rat hepatic Ito cells, a fibrogenic effector cell which proliferates in vivo during liver injury and fibrogenesis. D3 stimulated Ito cell DNA synthesis and potentiated platelet-derived growth factor-induced mitogenesis. D3's enhancement of [3H]thymidine incorporation was associated with nuclear Egr expression. Recent studies have causally linked the activated proto-oncogene c Raf with downstream Egr induction. The serine-threonine kinase Raf protein is phosphorylation-activated by a large array of agonists including plasma membrane and cytoplasmic tyrosine kinases but has not previously been associated with the steroid superfamily of mediators. To consider potential prenuclear acute pathways of D3-induced stimulation, the activation of Raf was examined following D3 exposure. D3 induced Raf activation as assessed via (a) enhanced Raf phosphorylation following in vivo 32P labeling, (b) enhanced kinase function utilizing exogenous histone 1 protein as substrate, and (c) the shift in Raf physical localization changing from a diffuse cytoplasmic distribution to a perinuclear domain. A similar activation of Raf kinase was found in 3T3 cells exposed to D3 with enhanced histone phosphorylation detectable within 1 min following stimulation. The proximal cascade leading to Raf kinase activation may involve a protein kinase activity was severely attenuated by stimulated kinase activity was severely attenuated by previous phorbol ester treatment for 20 h or staurosporine pretreatment. PMID- 8227077 TI - Independent regulation of two cytoplasmic processing stages of the intermediate filament-associated protein filaggrin and role of Ca2+ in the second stage. AB - One of the final events in cornification of epidermal cells is processing of profilaggrin to the keratin-associated protein filaggrin. Processing involves several proteolytic events and occurs in two discrete proteolytic stages (Resing, K. A., Walsh, K. A., and Dale, B. A. (1984) J. Cell Biol. 99, 1372-1378; Resing, K. A., Walsh, K. A., Haugen-Scofield, J., and Dale, B. A. (1989) J. Biol. Chem. 264, 1837-1846). In a keratinocyte cell line derived from newborn rat epidermis, these two stages are independently regulated. Profilaggrin was expressed when the cells reached confluence; processing to intermediates began 24-36 h later (stage one), with filaggrin appearing at 48 h (stage two). Stage two processing required calcium in the medium with maximum processing occurring at 5-10 mM. Furthermore, stage two processing was inhibited by nifedipine, a calcium channel blocker, suggesting that calcium influx activates this event. Second-stage processing was also inhibited by the protease inhibitor leupeptin, implicating calpain. Confluent cells had higher levels of calpain I than subconfluent cells; in confluent cells, two immunoreactive bands were detected, comigrating with inactive (80 kDa) and activated (78 kDa) calpain I. In cells processing profilaggrin, most of the calpain I was in the 78-kDa form, implying extensive activation, supporting a role for calpain in processing. PMID- 8227078 TI - Phosphorylation of synthetic peptides containing Tyr-Met-X-Met motifs by nonreceptor tyrosine kinases in vitro. AB - Several tyrosine phosphorylation sites in the insulin receptor kinase substrate IRS-1 are predicted to be within Tyr-Met-X-Met (YMXM) motifs, and synthetic peptides corresponding to these sequences are excellent substrates for the insulin receptor kinase in vitro (Shoelson, S. E., Chatterjee, S., Chaudhuri, M., and White, M. F. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 2027-2031). In this study, YMXM-containing peptides are shown to act as substrates for two members of the nonreceptor subfamily of tyrosine kinases, v-Src and v-Abl (the transforming gene products of Rous sarcoma virus and Abelson murine leukemia virus, respectively). For v-Src, a baculovirus expression system was used which was capable of producing milligram quantities of pure 60-kDa v-Src in Spodoptera frugiperda (Sf9) cells. The source of v-Abl was an Escherichia coli expression vector that produces a fusion protein of glutathione S-transferase with the abl catalytic domain. The synthetic YMXM-containing peptides had among the highest apparent affinities described to date for either tyrosine kinase, with Km values as low as 97 microM for v-Src and v-Abl. Comparisons with the results obtained with the insulin receptor kinase revealed differences in substrate specificity among the enzymes. In particular, v-Src was more tolerant of substitutions at the Met+1 and Met+3 positions in the YMXM motif than either v-Abl or the insulin receptor kinase but was more dependent on the presence of a preceding acidic amino acid. For v-Abl, the presence of threonine at any position in the YMXM motif caused a reduction in catalytic efficiency. Phosphorylated YMXM motifs are recognition elements for binding to the src homology 2 domains of phosphatidylinositol 3'-kinase and additional proteins; hence, differences in specificity of tyrosine kinases toward YMXM-containing proteins may have relevance to downstream signaling events. PMID- 8227079 TI - Molecular cloning of two abundant protein tyrosine kinases in Torpedo electric organ that associate with the acetylcholine receptor. AB - The nicotinic acetylcholine receptor (AChR) is phosphorylated on tyrosine residues both in vitro and in vivo. This phosphorylation appears to regulate the rate of receptor desensitization and is associated with AChR clustering induced by the neuronal extracellular matrix protein agrin. To identify the protein tyrosine kinase(s) that phosphorylates the nicotinic receptor, we have used molecular cloning techniques to identify and characterize two protein tyrosine kinases that are highly expressed in Torpedo electric organ, a tissue enriched in synaptic components including the AChR. One of the kinases was identified as the Torpedo homolog of neuronal fyn, whereas the other was a novel kinase we have named fyk due to its homology to both fyn and yes protein tyrosine kinases. Using antibodies to fyn and fyk, Fyn was shown to be a 55-kDa protein phosphorylated on tyrosine residues, whereas Fyk was a 56-kDa/53-kDa doublet phosphorylated on serine and tyrosine residues. At the mRNA and/or protein level, fyn and fyk were present in Torpedo electric organ, skeletal muscle, and brain. Both kinases were detected in the membrane fractions enriched in the AChR, with Fyn and Fyk representing 36 and 8%, respectively, of the protein tyrosine kinase activity in these postsynaptic membranes. In addition, Fyn and Fyk were shown by coimmunoprecipitation to be specifically associated with the AChR. Furthermore, the AChR was phosphorylated in Fyn and Fyk immunoprecipitates. These results indicate that Fyn and Fyk are involved in the regulation of postsynaptic membrane function and suggest that these protein tyrosine kinases may phosphorylate the AChR. PMID- 8227080 TI - Movement of apolipoprotein B into the lumen of microsomes from hepatocytes is disrupted in membranes enriched in phosphatidylmonomethylethanolamine. AB - When monolayer cultures of rat hepatocytes are incubated with the ethanolamine/choline analogue, monomethylethanolamine, the secretion of apolipoproteins B100 and B48, as well as the lipid constituents, of very low density lipoprotein (VLDL) is inhibited by approximately 50% (Vance, J. E. (1991) J. Lipid Res. 32, 1971-1982). In the present study we have investigated the mechanism by which monomethylethanolamine disrupts VLDL secretion. Hepatocytes were treated with 400 microM monomethylethanolamine overnight, which resulted in an increase in the cellular content of the derived phospholipid, phosphatidylmonomethylethanolamine, from 0.32 +/- 0.15 to 2.92 +/- 0.74 nmol/mg of cell protein. The biosynthesis of apoproteins B100 and B48 was not impaired by treatment of cells with monomethylethanolamine. However, monomethylethanolamine decreased by approximately 50% the amount of apoproteins B, but not of the typical secretory protein, albumin, present in the luminal content subfraction of microsomes. The intracellular degradation of apoproteins B was also increased in phosphatidylmonomethylethanolamine-enriched, compared with control, cells. Moreover, the pool of apoprotein B present in intact microsomes from hepatocytes incubated with monomethylethanolamine was more accessible to exogenously added trypsin, presumably because a larger pool of the apoprotein B was exposed on the cytosolic surface of these microsomes. The data strongly suggest that an increase in the microsomal content of phosphatidylmonomethylethanolamine diminishes the ability of apoprotein B to translocate across the endoplasmic reticulum membrane into the luminal compartment. Consequently, the association of apoprotein B with lipids and/or the normal assembly of mature VLDL particles is impaired. PMID- 8227081 TI - Characterization of the lectin from the bulbs of Eranthis hyemalis (winter aconite) as an inhibitor of protein synthesis. AB - The lectin from Eranthis hyemalis has been previously characterized as consisting of two polypeptide chains covalently linked by disulfide bond(s) (Cammue, B. P., Peeters, B., and Peumans, W. J. (1985) Biochem. J. 227, 949-955). We have further characterized the biochemical properties of the lectin and demonstrated that it possesses the property of inhibition of protein synthesis using in vitro eukaryotic translation systems. The protein also possesses antiviral activity against the plant virus, alfalfa mosaic virus, and larvicidal activity against the southern corn rootworm, Diabrotica undecimpunctata howardii, a major insect pest of the maize plant. Both isoelectric focusing on gels and chromatofocusing indicated heterogeneity of the protein, with three species having isoelectric points in the range 4-5. The disulfide bond(s) can be rapidly reduced with beta mercaptoethanol under native conditions. The reduced alkylated polypeptide chains remain associated under native conditions to form a species, EHL', that elutes at the same position as the native protein and has the same molecular weight by sedimentation equilibrium experiments. However, circular dichroism and fluorescence measurements indicated conformational differences between the species. PMID- 8227082 TI - Purification of a 110-kDa phosphoinositide phospholipase C that is activated by G protein beta gamma-subunits. AB - We report the purification from bovine brain cytosol of a 110-kDa phosphoinositide-specific phospholipase C (PLC-110) that was markedly stimulated by G-protein beta gamma-subunits. The enzyme was purified approximately 2000-fold with a yield of 4%. On the basis of size and immunological cross-reactivity, PLC 110 was distinct from 150-kDa PLC-beta 1, 145-kDa PLC-gamma 1, and 85-kDa PLC delta 1. An antiserum to a peptide corresponding to a conserved PLC Y domain sequence cross-reacted with PLC-110. PLC-110 was also recognized by two antisera selective for NH2-terminal and internal sequences in PLC-beta 3, but not by a third peptide antiserum to the COOH terminus of this enzyme, suggesting that PLC 110 is related to PLC-beta 3. Reconstitution of purified PLC-110 with beta gamma subunits produced greater than 100-fold activation, indicating activation was observed at approximately 60 nM beta gamma and full activation at approximately 500 nM beta gamma. PLC-110 maximally hydrolyzed phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate at 1 microM Ca2+, but showed no activity toward phosphatidylinositol at Ca2+ concentrations up to 1 mM. Concentrations of purified guanosine 5'-O-(3-thiotriphosphate)-liganded alpha q that fully activated PLC-beta 1 failed to stimulate PLC-110. This observation indicates that the site at which beta gamma interacts with PLC-110 is distinct from that at which alpha q regulates the activity of PLC-beta isozymes. PMID- 8227083 TI - Function of the GrpE heat shock protein in bidirectional unwinding and replication from the origin of phage lambda. AB - The initiation of DNA replication by phage lambda depends on a specialized nucleoprotein structure that provides for the precise localization and activity of the Escherichia coli DnaB helicase at the lambda replication origin. Previous work has shown that the DnaJ and DnaK heat shock proteins function in the initiation pathway by releasing the DnaB helicase from the initiation complex to carry out localized unwinding of origin DNA. This DnaJ.DnaK pathway results in mainly unidirectional DNA unwinding and replication, whereas replication in vivo is mainly bidirectional. Based on recent replication work indicating an important role for the GrpE heat shock protein, we have used electron microscopy to study the action of GrpE in the DNA unwinding and replication reactions. We have found that GrpE acts with DnaJ and DnaK to facilitate the unwinding reaction at low concentrations of DnaK. In the presence of GrpE, bidirectional unwinding occurs in approximately half of the unwound DNA molecules. In addition, GrpE significantly increases the frequency of replication proceeding leftward from the origin. We suggest that reactions including GrpE result in more complete disassembly of the preinitiation nucleoprotein structure, thus allowing replication to proceed in both directions from the origin. PMID- 8227084 TI - Osteogenic protein-1 regulates L1 and neural cell adhesion molecule gene expression in neural cells. AB - Osteogenic protein-1 (OP-1) is a member of the TGF-beta superfamily that is expressed in the nervous system. We recently showed that human recombinant osteogenic protein-1 (hOP-1) strongly promotes the aggregation of dividing neuroblastoma x glioma hybrid NG108-15 cells, in part by inducing the major isoforms of the neural cell adhesion molecule (N-CAM) (Perides, G., Safran, R. M., Rueger, D. C., and Charness, M. E. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 10326-10330). Here we show that hOP-1 induces L1 expression approximately 6-fold in NG108-15 cells without changing the levels of N-cadherin, neurofilament 200, Thy-1, tau, and G alpha s. OP-1 induction of L1 and N-CAM was unassociated with changes in cell proliferation and was not reproduced by cellular differentiation. The increased adhesiveness of hOP-1-treated NG108-15 cells could be inhibited in part by Fab fragments of an anti-L1 polyclonal antiserum. L1 and N-CAM expression first increased 12-18 h after hOP-1 treatment, reached a maximum after 2-3 days, persisted for up to 5 days, and returned to control levels 3 days after hOP-1 withdrawal. The increases in L1 and N-CAM protein levels were preceded or accompanied by large increases in the abundance of L1 and all detectable N-CAM mRNAs. Actinomycin D prevented the induction by hOP-1 of L1 and N-CAM mRNAs, suggesting that hOP-1 regulates immunoglobulin CAM gene transcription. OP-1 is the first described growth factor that regulates both N-CAM and L1 gene expression. PMID- 8227085 TI - Loading dependence of inositol 1,4,5-trisphosphate-induced Ca2+ release in the clonal cell line A7r5. Implications for the mechanism of quantal Ca2+ release. AB - The Ca2+ content of the intracellular Ca2+ stores controls the inositol 1,4,5 trisphosphate receptor (InsP3R) in the clonal cell line A7r5. This regulation was characterized with respect to the understanding of the "quantal" release phenomenon. Independent of the loading protocol used, increasing the Ca2+ content of the stores increased the sensitivity of the inositol 1,4,5-trisphosphate (InsP3)-induced Ca2+ release until a Ca2+ content of about 30% of the steady state value was reached. Loading of the stores to higher levels had only a marginal effect on the Ca2+ release. The effects of luminal Ca2+ were still observed in the presence of 10 mM BAPTA (1,2-bis(O-aminophenoxy)ethane-N,N,N',N' tetraacetic acid), excluding the possibility that luminal Ca2+ acted indirectly via cytoplasmic binding sites. Conditions were developed to simultaneously measure [3H]InsP3 binding to the InsP3R and the 45Ca2+ content of the stores in the same cells. [3H]InsP3 binding to a high affinity binding site was potentiated by luminal Ca2+. Analysis at the molecular level revealed the simultaneous expression of different splice variants of InsP3R-I, as well as the expression of InsP3R-III, and of the putative InsP3R-IV. We conclude that the control of the InsP3R by luminal Ca2+ could account for quantal release and that the observed heterogeneity of the InsP3R may also contribute to this behavior, especially at high levels of store loading. PMID- 8227086 TI - Calmodulin-cardiac troponin C chimeras. Effects of domain exchange on calcium binding and enzyme activation. AB - Calmodulin (CaM) and the cardiac isoform of troponin C (cTnC) are close structural homologs, but cTnC cannot activate most CaM target enzymes. To investigate structure-function relationships, we constructed a series of CaM.cTnC chimeras and determined their ability to bind Ca2+ and activate CaM target enzymes. Previously, we exchanged domain 1 and found that the chimeras exhibited profoundly impaired activation of smooth muscle myosin light chain kinase (smMLCK) and had differential effects on other CaM target enzymes (George, S. E., VanBerkum, M. F. A., Ono, T., Cook, R., Hanley, R. M., Putkey, J. A., and Means, A. R. (1990) J. Biol. Chem. 265, 9228-9235). One of the domain 1 chimeras was a potent competitive inhibitor of smMLCK. We now extend our study of CaM.cTnC chimeras by exchanging all of the remaining functional domains of CaM and cTnC. We determined the ability of the chimeras to bind Ca2+ and activate phosphodiesterase (PDE) and smMLCK. Chimeras containing both domains 3 and 4 of cTnC exhibited high affinity Ca2+ binding that was indistinguishable from cTnC, whereas chimeras containing either domain 3 or 4 of cTnC demonstrated Ca2+ affinity that was intermediate between CaM and cTnC. All of the CaM.cTnC chimeras showed near-maximal PDE activation but required 5-775-fold higher concentrations than CaM to produce half-maximal PDE activation. In contrast, all of the chimeras showed impaired ability to activate smMLCK, and some were potent competitive inhibitors of smMLCK activation by CaM. PMID- 8227087 TI - The role of nucleotides conserved in eukaryotic initiator methionine tRNAs in initiation of protein synthesis. AB - Mutant human initiator tRNA genes carrying changes in each of the three features unique to eukaryotic initiator tRNAs have been constructed, and introduced into CV-1 monkey kidney cells using SV40 virus vectors. The mutant tRNA genes are expressed, and the mutant tRNAs can all be aminoacylated with both rabbit liver and Escherichia coli methionyl-tRNA synthetases. Based on aminoacylation levels, the tRNAs are expressed to 5-15-fold over the level of endogenous initiator tRNA. The activity of the mutant [35S]methionyl-tRNAs in initiation was studied in rabbit reticulocyte and wheat germ cell-free protein synthesis systems programmed with various mRNAs. Initiation is studied by using a mRNA that codes for a protein whose N-terminal methionine is stable and not removed by methionine aminopeptidase. Changing the A1:U72 base pair to a G1:C72 base pair greatly reduced activity of the tRNA in initiation. Changing the three consecutive G:C base pairs (G29G30G31:C39C40C41) in the anticodon stem to those found in elongator methionine tRNA also reduced initiation activity. Interestingly, changing the A54 and A60 residues in loop IV to T54 and U60 had less of an effect on activity. The tRNA with changes in all three conserved features had virtually no activity in initiation. PMID- 8227088 TI - Stable fluorescent dye-DNA complexes in high sensitivity detection of protein-DNA interactions. Application to heat shock transcription factor. AB - The gel mobility-shift assay is an important tool for the study of protein nucleic acid interactions. High detection sensitivity is typically attained by radioisotopic labeling of the target nucleic acid fragments. A novel fluorescence methodology offers significant advantages over this conventional approach. Ethidium, thiazole orange, and oxazole yellow homodimers form stable, highly fluorescent complexes with double-stranded DNA that can be detected in gels by a laser-excited, confocal, fluorescence scanning system with a sensitivity higher than that attainable with radioisotopic labeling. We describe here the use of these dyes in a gel-mobility assay to detect complexes of a truncation of the Kluyveromyces lactis heat shock transcription factor, containing the trimerization and DNA-binding domains (HSFDT), with target DNA. At an appropriate molar DNA base pair to dye ratio, the labeling of a DNA fragment with dimeric dye did not affect the binding to HSFDT. The detection of the fluorescent-dye labeled HSFDT-DNA complexes with the laser scanner achieves a spatial resolution far superior to that of conventional autoradiography and permits analysis of multimer protein-DNA complexes that are not resolved by traditional detection methods. We have used this technique to demonstrate that HSF forms multimeric complexes on DNA by addition of trimeric units. The latter conclusion is based on an analysis of the mobilities of the multiple HSFDT-DNA complexes and on a two-color mobility shift fluorescence assay that uses a mutant of HSFDT engineered for site-specific labeling with fluorescein and target DNA labeled with an "energy transfer" dye, thiazole orange-thiazole blue heterodimer. PMID- 8227089 TI - Spatial resolution of fodrin proteolysis in postischemic brain. AB - One of the major obstacles in investigating in vivo proteolytic phenomena has been the inaccessibility to spatial information as to where in the tissue the reaction proceeds because the orthodox method employing electrophoretic analysis requires homogenization of samples and thus results in loss of such spatial information. To overcome this technical drawback, we have developed methodology to produce antibodies that specifically distinguish a proteolyzed form of a given protein from its intact form. Here we describe our immunohistochemical observation of calpain-catalyzed fodrin proteolysis in postischemic gerbil hippocampus, using an antibody exclusively specific to the proteolyzed 150-kDa form of fodrin alpha subunit. Our data establish a novel discovery that transient (10 min) global forebrain ischemia followed by reperfusion induces at least two distinct phases of fodrin proteolysis in hippocampus: an early phase in molecular layer and in stratum oriens of CA3 and CA1 sectors within 15 min and a late drastic and persistent phase in the entire CA1 after 4-24 h. The former may be one of the early events initiating the complex cascade leading to the delayed neuronal death, while the latter should be considered as a more direct cause for the actual degeneration in CA1. PMID- 8227090 TI - In vitro chamber specification during embryonic stem cell cardiogenesis. Expression of the ventricular myosin light chain-2 gene is independent of heart tube formation. AB - The molecular cues that control patterning of the heart tube during early cardiogenesis are largely unknown. The present study has explored the embryonic stem (ES) cell differentiation system to determine if this in vitro model could be useful in studying the process of regional specification of cardiac muscle cells at the earliest possible stages. As assessed by polymerase chain reaction, ribonuclease protection, in situ hybridization, and immunohistochemical analyses, ES cell differentiation into embryoid bodies is characterized by the transcriptional and translational activation of the ventricular regulatory (phosphorylatable) myosin light chain gene, demonstrating that ventricular specification occurs during ES cell cardiogenesis. The finding of a ventricular specific marker in an in vitro system in the absence of an intact heart tube provides evidence for cardiac regional specification independent of positional cues or physiologic stimuli. The temporal expression of the myogenic regulatory factors, myogenin and MyoD, suggests activation of the skeletal muscle program following cardiac myogenesis in vitro, indicating temporal fidelity to the progression of in vivo myogenesis. These data establish the mouse embryonic stem cell system as a model for cardiac chamber specification and suggest a promising approach in the study of regional specification in genetically engineered cardiac muscle cells. PMID- 8227091 TI - Characterization and chromosomal mapping of the human thromboxane A2 receptor gene. AB - The human thromboxane A2 gene, present as a single copy, spans over 15 kilobases (kb) and contains 3 exons divided by 2 introns. The first intron, intron 1, exists in the 5'-noncoding region, 83 base pairs upstream from the ATG start site and is 6.3 kb long. Intron 2 with a length of 4.3 kb is located at the end of the sixth transmembrane region, thereby separating it from the downstream coding sequences including the seventh transmembrane region and the 3'-untranslated region. By rapid amplification of 5'-cDNA ends, transcription initiation sites starting in two different putative promoter regions were determined. In the 5' flanking region of these transcription initiation sites, no typical TATA box exists. The major promoter, the promoter region I, contains four potential SP-1 binding sites and several potential binding sites for other transcription factors. By polymerase chain reaction analysis, a small portion of the RNA transcribed from this gene was shown to contain an additional 115-base pair long noncoding exon, exon 1b, which is located in intron 1. No additional exon in intron 2 was detectable, indicating that a single type of thromboxane A2 receptor protein is encoded by this gene. Chromosomal localization was carried out by fluorescence in situ hybridization of cloned genomic DNA to the metaphase chromosome. The gene was assigned to 19p13.3 of human chromosome. PMID- 8227092 TI - Attenuation of nursing-related ovarian suppression and high fertility in well nourished, intensively breast-feeding Amele women of lowland Papua New Guinea. AB - Intense, sustained nursing lengthens inter-birth intervals and is causally linked with low natural fertility. However, in traditional settings, the effects of such nursing on fertility are difficult to disentangle from those of nutrition. Results from a prospective, direct observational study of reproductive function in well-nourished Amele women who nurse intensively and persistently but who also have high fertility are here presented. Endocrine measures show that ovarian activity resumes by median 11.0 months postpartum. Median duration of postpartum amenorrhoea is 11.3 months, time to next conception is 19.0 months, and the inter birth interval is 28.0 months. Average life time fertility is 6.8. High fertility in Amele women is due both to refractoriness of reproductive function to suckling stimuli, and to maintenance of equivalent age-specific fertility rates across the reproductive life span. PMID- 8227093 TI - Does women's literacy affect desired fertility and contraceptive use in rural urban Pakistan? AB - The 1984-85 Pakistan Contraceptive Prevalence Survey showed that urban wives had more than twice the literacy rate of rural wives. The present study explored the relationship of the rural-urban gap in female literacy to differences in contraceptive use. In rural areas, literacy did not increase women's perceptions of having reached a 'sufficient' number of living children, although the opposite was true for urban areas. Yet rural women with an 'insufficient' number of living children were more likely to use contraception if they were literate, as did their urban counterparts. Thus, raising the literacy rate in rural Pakistan would not narrow the rural-urban gap in contraception to cease childbearing but would narrow the rural-urban gap in contraception used to space wanted births further apart. Recommendations for government policy are made. PMID- 8227094 TI - Effects of family sex composition on fertility preference and behaviour in rural Bangladesh. AB - The effects of family sex composition on fertility preferences and behaviour during the period 1977-88 are examined using longitudinal data from Matlab, Bangladesh. The sex composition of living children was found to be systematically related to fertility preferences and behaviour, with a higher number of sons at each family size associated with a higher percentage of women wanting no more children, a higher percentage currently using contraception, and lower subsequent fertility. However, the highest percentage wanting no more children, using contraception and having lowest subsequent fertility was found among women who already had one daughter as well as at least one son. The effects of sex preference on fertility preferences and behaviour were measured using an index developed by Arnold (1985). The results suggest that while sex preference remained largely unchanged during the study period, its effect on contraceptive use declined and its impact on actual fertility remained modest and fairly stable. PMID- 8227095 TI - Premature childbirth: social and behavioural risks in Singapore. AB - The associations of social and behavioural factors on preterm birth in Singapore were studied using hospital maternity records of 20,723 consecutive singleton births between January 1986 and November 1991. The overall proportion of preterm births was 3.6%, the rates for the Chinese, Malay and Indian groups being 3.2%, 3.8% and 4.9% respectively. Teenage mothers were at a higher risk of preterm labour compared to women aged 20-29 years. The incidence of preterm labour decreased with increasing educational status. Preterm births were six times more likely in women who had no antenatal care. Women who had three or more previous births were at a higher risk, while those who had one or two previous births were at a lower risk compared to women who had none. PMID- 8227096 TI - The polygyny-fertility hypothesis revisited: the situation in Ghana. AB - Re-examination of the polygyny-fertility hypothesis, by multiple regression analysis of the 1988 Ghana DHS data, reveals that polygyny has no effect on the number of children ever born; i.e. polygynously married women are as fertile as those monogamously married. Husband's age has no significant role in determining the number of children ever born to a woman. PMID- 8227097 TI - Urban nuptiality patterns and marital fertility in Nigeria. AB - This study indicates that urban marital patterns in nine Nigerian cities influence fertility. Fertility is also influenced by age at marriage, region of residence, ethnicity and religion; education and employment lead to marriage delay and tend to conflict with childbearing by enhancing the status of women. PMID- 8227098 TI - Socioeconomic factors affecting marriage, divorce and birth rates in a Japanese population. AB - The effects of low income, urbanisation and young age population on age-adjusted rates of first marriage, divorce and live birth among the Japanese population in 46 prefectures were analysed by stepwise regression for 1970 and for 1975. During this period, Japanese society experienced a drastic change from long-lasting economic growth to serious recession in 1973. In both 1970 and 1975, the first marriage rate for females was inversely related to low income and the divorce rates for both males and females were positively related to low income. The live birth rate was significantly related to low income, urbanisation and young age population only in 1975. The first marriage rate for females and the divorce rates for both sexes increased significantly but the first marriage rate for males and live birth rate significantly decreased between 1970 and 1975. These findings suggest that low income was the essential factor affecting first marriage for females and divorce for males and females. PMID- 8227099 TI - Effect of son preference and population policy on sex ratios at birth in two provinces of China. AB - Two samples of births were used to examine the effect of son preference and population policy on sex ratios in Hebei and Shaanxi provinces of China. The results from all births to women aged 15-49 in the 1985 In-Depth Fertility Survey indicated a strong son preference but no clear policy impact on sex ratios at different birth orders. In a sample of selected births during 1975-84 to women aged 40 or under, the mixed influences of son preference and population policy were clearly reflected in the high and rising overall ratios at birth, and the increasing sex ratios with parity. Possible explanations for these findings are discussed. PMID- 8227100 TI - Marriage distances among the Afroamericans of Bluefields, Nicaragua. AB - In a sample of 311 couples from the Afroamerican community of Bluefields, Eastern Nicaragua, the distribution of matrimonial distance shows a deviation from the leptokurtic rule. This results from assortative mating among the population. PMID- 8227101 TI - Menarcheal age in University of Warwick young women. AB - Data from the final 16 years of a 28-year ongoing survey of menarcheal age are reported. From 1971 onwards, recalled aged at menarche was recorded for all young women entering the University of Warwick. These data show that mean menarcheal age is increasing, a trend which is independent of father's occupation, family size, position of girl in the family, and physique. This continues a similar upward tendency noted in a preceding study in Swansea. It appears that the downward trend to earlier age at menarche of the earlier decades of this century has been replaced by one in the opposite direction. PMID- 8227102 TI - Socioeconomic status: the prime indicator of premature death in Australia. AB - The link between socioeconomic status and health has long been recognised. This study of deaths among Australian men aged 15-59 years demonstrates that during the 20-year period, 1966-86 the number of premature deaths was dramatically reduced among all socioeconomic groups, primarily as a result of falls in death rates due to heart disease, stroke and trauma. However, the marked differences in death rates according to social class remain, to the extent that if men of all social classes had the same mortality experiences as professional and technical workers the overall death rates for Australian men would be reduced by 60%. Socioeconomic status is the most important indicator of health status among Australians. PMID- 8227103 TI - Consanguinity in north Jordan: prevalence and pattern. AB - The rate of consanguineous mating in north Jordan was 63.7% with an average inbreeding coefficient of 0.0283. First cousin marriages were the most predominant type (58.5%); villagers showed higher levels than city dwellers. The level of female education did not influence consanguinity rate. PMID- 8227105 TI - Abstracts of papers presented at recent scientific meetings organised by orthopaedic associations and specialist societies. 1991-1993. PMID- 8227104 TI - Sex differences in primary cognitive abilities among blacks, Indians and whites in South Africa. AB - Sex differences are reported for samples of approximately 1000 16-year-old blacks, Indians and whites in South Africa on ten tests of cognitive ability. Males obtained significantly higher means on non-verbal reasoning, spatial and mechanical aptitude, and females obtained significantly higher means on perceptual speed and memory for meaning (except among the black sample). In general the sex differences in South Africa are consistent with those typically obtained in the United States. PMID- 8227106 TI - High porosity PTFE improves endothelialization of arterial grafts without increasing early thrombogenicity. AB - OBJECTIVE: High porosity (HP) (90 micron internodal distance) PTFE grafts were implanted into the carotid and femoral arteries of dogs to investigate early thrombogenicity, patency, and endothelialization. EXPERIMENTAL DESIGN: Standard PTFE (STD) grafts (30 micron internodal distance) were used as controls. 12 HP and 12 STD grafts were implanted into 6 dogs. Indium-111 labeled platelets were infused intravenously after graft implantation. A graft platelet accumulation index (GPAI) was calculated as the ratio of radioactive emission from the PTFE grafts excised at 48 hours to the emission from a native arterial segment. Another 12 HP and 12 STD grafts were implanted into femoral and carotid arteries to assess patency and endothelialization at 4 and 18 weeks. RESULTS: There was no significant difference in the GPAI of the HP and STD grafts at either the carotid (HP = 31.5 +/- 9.7, STD = 30.6 +/- 11.8; p > 0.05) or femoral (HP = 34.0 +/- 5.0, STD = 31.5 +/- 7.9; p > 0.05) positions. Combined data (carotid and femoral HP vs. carotid and femoral STD) also did not demonstrate a difference in GPAI (HP = 32.8 +/- 7.5, STD = 31.1 +/- 9.6; p > 0.05). Patency rates were the same at 4 weeks (75%), but greater in the HP grafts at 18 weeks (HP = 75%, STD = 37%; p > 0.05). No difference was noted in the percentage of graft endothelialization at 4 weeks (HP = 5.2 +/- 5.8, STD = 5.0 +/- 4.0; p > 0.05), however, at 18 weeks the HP graft had significantly more endothelial coverage compared to STD grafts (HP = 75.2 +/- 13.9, STD = 22.6 +/- 9.5; p < 0.01). CONCLUSIONS: Given that HP PTFE is no more thrombogenic than STD PTFE, and that it provides superior endothelialization, HP grafts should continue to be developed and studied for potential clinical use. PMID- 8227107 TI - Ultrasonic classification of carotid plaques causing less than 60% stenosis according to ultrasound morphology and events. AB - Carotid plaque morphology studied by ultrasound has been used to classify 3 types of bifurcation plaques, producing less than 60% stenosis on duplex scanning: Type A, the homogeneous, highly echogenic plaque; Type B, the heterogeneous (mainly echogenic, B-1 or mainly echolucent, B-2) plaque; Type C, the complex plaque with irregular surface, echogenic and/or echolucent with calcifications. One hundred asymptomatic subjects in each plaque group were included in a prospective 4-year follow-up study to evaluate clinical events and brain damage by yearly cerebral CT and MR scans. After four years 96 subjects with type A plaque, 89 with type B 1 and 67 with type C plaques completed the study. While there were no clinical events or positive cerebral scans in subjects with type A plaques there was a significantly increasing number of cerebral and vascular events in subjects with plaque B and C type with a zero mortality in the plaque A group, a 2.24% mortality in type B plaques and a significantly higher (p > 0.05) mortality (13.4%) in type C plaques. CT and MR scans revealed brain damage which had been totally asymptomatic with MR detecting a larger number of lesions. Also the number of cerebral lesions observed by CT/MR were significantly greater than the number of recorded clinical episodes. ECG stress test revealed a larger number of asymptomatic coronary disease in the C type plaques. In conclusion ultrasound plaque morphology is linked to different incidences of cerebral and cardiac events, brain damage and asymptomatic coronary disease. Also CT and MR may reveal lesions which do not cause signs or symptoms. PMID- 8227108 TI - The functional long-term value of different types of treatment for saphenous vein incompetence. AB - OBJECTIVE: To compare the long-term value of different forms of treatment for primary varicose veins with saphenous vein insufficiency. EXPERIMENTAL DESIGN: A prospective, partially randomized study with 5-year follow-up. SETTING: Ambulatory day-case care. PATIENTS AND INTERVENTIONS: The study includes 211 patients (214 lower limbs), who received compression sclerotherapy (CST; n = 78), radical operation (OP; n = 74) or CST combined with high tie under local anesthesia (HT + CST; n = 63). MEASURES: The patient's subjective opinion, objective finding by the surgeon and functional (foot-volumetric) assessment were obtained just after treatment and 6 months, 1, 3 and 5 years later. RESULTS: Subjectively the result started to deteriorate in both the CST and HT + CST groups after one year. The patient satisfaction was greatest in the OP group throughout the study period. Objectively the CST group cure rate fell markedly after 6 months and at 5 year follow-up the failure rate reached 51%, while the OP group still had a high rate of cured (60%) and improved (35%) patients. The HT + CST treatment seemed to hold well for three years followed by increasing failure rate with only 16% objectively cured after 5 years. The foot-volumetric parameters expelled volume (= calf pump function) and refilling flow ratio (= venous reflux) increased 51-79% respectively decreased 8-29% post-treatment in all groups. After 5 years these parameters had returned to pre-treatment levels in the CST and HT + CST groups, while the OP group was still significantly improved. CONCLUSIONS: Radical surgery is superior to compression sclerotherapy alone or in combination with high tie in the treatment of varicose veins with saphenous vein incompetence. The foot-volumetric assessment correlated well with and supported objective findings as a whole but could not replace the clinical examination of each individual patient. PMID- 8227109 TI - Femoropopliteal and femorotibial greater saphenous vein "in situ" reconstructions in non selected patients. Life table analysis. AB - 594 non selected "in situ" reconstructions were analyzed retrospectively using the life table method. Especially after immediate occlusion the cumulative patency rate is unsatisfactory (33.1% vs 62.3% after 6 years). Patency rates are influenced by the anastomotic site and mainly by run off conditions. The number of patent tibial arteries seems to be the most important determinant. These results prove to be statistically significant. PMID- 8227110 TI - Acute extensive necrosis of the visceral organs after repair of a ruptured thoracic aortic aneurysm. AB - A 47-year-old woman was admitted to our hospital for repair of a ruptured thoracic aortic aneurysm. Her post-operative course was uneventful, but she had persistent complaints of anorexia and general fatigue that failed to improve. Thirty-one days after surgery, she complained of severe abdominal pain. Abdominal computed tomography (CT) and X-ray revealed extensive necrosis of the visceral organs. She underwent emergency abdominal exploration and was found to have necrosis of the liver, gallbladder, stomach, entire small bowel and colon. The extensive necrosis made resection of the involved organs unfeasible. The patient died one day after exploratory laparotomy. PMID- 8227111 TI - Surgical workshop PTFE "gutter" angioplasty. PMID- 8227112 TI - Delayed arteriosclerosis progression in high risk subjects treated with mesoglycan. Evaluation of intima-media thickness. AB - Noninvasive ultrasonic biopsy (UB) can be used to classify arteriosclerotic lesions and their progression in the carotid and femoral bifurcation. Also the evaluation of intima-media thickness (IMT) is useful to quantify the progression of early arteriosclerosis. Two randomly selected groups of asymptomatic subjects were included in a 18 month, open study. One group was treated with oral mesoglycan (200 mg/day) and one group was followed-up as control. The two groups were comparable for age and sex distribution. The average UB score was 14.4 +/- 5 in the treatment group and 14.3 +/- 8 in the control group. After 18 months the UB score was 15.7 +/- 4 in the treatment and 16.2 +/- 6 in the control group. These differences were not significant. However the average increase in IMT in 18 months in the treatment group was 0.016 mm equivalent to 0.0106 mm per year. In the control group the average increase was 0.119 equivalent to 0.0793 per year. Therefore the increase in IMT was 7.48 times greater in the control group. These differences were significant (p < 0.05). Two drop-outs were recorded in the treatment group and 1 in the control group. In conclusion IMT measurements showed a decreased level of IMT progression in subjects under mesoglycan treatment. These results need to be confirmed by a larger randomised study. PMID- 8227113 TI - Perioperative arrhythmia associated with aortic valve stenosis. AB - Peri-operative arrhythmia is one of the major complications in anaesthesia for valve replacement surgery in patients with aortic stenosis. In this retrospective study, 58 patients with sinus rhythm were investigated from induction of anaesthesia until arrival at the recovery room by close haemodynamic monitoring and Holter ECG recording. After cardiopulmonary bypass (CPB), they received either lidocaine (L, n = 35) or mexiletine (M, n = 23) via infusion for 24 hours. Pre-bypass incidence was 14% for supraventricular (SPBs) and 19% for ventricular serious arrhythmia (VPBs), i.e. high-grade forms which indicate possible deterioration and may require therapy (for all arrhythmia, incidences were 45 resp. 28%). VPBs was independently related to impaired left ventricular function (11 patients) and preoperative digitalis therapy (20 patients) but not to severity of stenosis, serum concentration of potassium (between 3.3 and 5.2 meq/l), or any other clinical parameters. Post-bypass incidence was SPBs 11% and VPBs 33%, the latter representing a significant increase compared to the first period (p < 0.03)--(all arrhythmia: 26 resp. 40%). VPBs was related to the need for multiple therapy including catecholamines and antiarrhythmic agents other than L or M, but no longer to preoperative parameters nor duration of intraoperative ischaemia. Incidences of arrhythmia for L and M were identical. While in these patients digitalis therapy may account for arrhythmia also in general anaesthesia, in valve replacement there is a post-bypass increase in VPBs which is not fully explained. Since the incidence is 33% in spite of anti arrhythmic therapy, both administered class IB drugs may not be the best therapeutic approach. PMID- 8227114 TI - Clinical study of 47 patients with reoperation after open mitral commissurotomy. AB - Mitral lesion in 47 patients who received reoperation after an initial open mitral commissurotomy (OMC) for mitral stenosis (MS) was studied. The diagnosis before OMC consisted of pure MS in 38 patients, with mild mitral regurgitation (MR) in 8 and with moderate MR in 1. At OMC the diameter of the mitral orifice was a mean of 1.3 +/- 0.5 cm and a mobile cusp was found in 12 and a thickened cusp in 35 patients. Subvalvular fusion was seen in 15 patients, calcification of cusp or subvalvular region in 10 and left atrial thrombus in 6. After OMC, the diameter of the mitral orifice was opened to a mean of 3.2 +/- 0.3 cm. The interval between the initial operation and the time of reoperation was a mean of 124 months. The mitral lesion necessitating reoperation was MS in 18, moderate to severe MR in 10, and MSR in 19. At reoperation the mitral orifice ranged in diameter from 0.8 to 4.2 cm (mean, 2.3 +/- 0.8 cm). There was no correlation between the occurrence of MR or restenosis and the subvalvular division at initial OMC. Left atrial thrombus was identified in four and calcification in 17 patients. At reoperation 46 patients received mitral valve replacement, while only 1 received a second OMC. The removed valve showed no pathological findings of active rheumatic lesion but more severe calcification and hyalinization of the valve than those at initial operation. There were three early deaths (6.4%) after reoperation. The recurring mitral lesion seemed to consist of deteriorated pathological changes of the valve and MR and thus mitral valve replacement was recommended to provide a favorable operative result for non-flexible lesion after OMC. PMID- 8227115 TI - Prospective evaluation of clinical scoring systems in infants and children with cardiopulmonary insufficiency after cardiac surgery. AB - OBJECTIVE: To document severity of illness and to evaluate the predictive value of clinical scoring systems in infants and children with cardiopulmonary insufficiency after cardiac surgery. DESIGN: Prospective study with follow up to hospital discharge. SETTING: A multidisciplinary pediatric ICU in a University Hospital. PATIENTS: Between 1/1989 and 4/1992 441 infants and children with congenital heart disease underwent open heart surgery. 128 of these patients developed postoperative cardiopulmonary insufficiency and were entered into this study. METHODS: Data relevant to the Acute Physiologic Score for Children (APSC), Pediatric Risk of Mortality (PRISM), Therapeutic Intervention Scoring System (TISS) and Organ System Failure (OSF) score were collected in all patients during the first 4 days of postoperative intensive care. RESULTS: The mean age of the patients was 1.5 +/- 0.2 years. The mean duration of mechanical ventilation and ICU care was 6.2 +/- 0.6 and 8.1 +/- 0.7 days, respectively. On the first postoperative day the mean APSC and PRISM scores of survivors and nonsurvivors were 13.9 +/- 1.3 vs 24.5 +/- 1.3 (p < 0.001) and 6.1 +/- 0.5 vs 19.6 +/- 1.9 (p < 0.001), respectively. The mean TISS and OSF scores of survivors and nonsurvivors were 46 +/- 0.8 vs 57.8 +/- 1.4 (p < 0.001), and 2.2 +/- 0.2 vs 3.4 +/- 0.2 (p < 0.001), respectively. The overall hospital mortality rate was 9.9%, the hospital mortality rate of patients with postoperative cardiopulmonary insufficiency 34%. Patients with an APSC score < 10 and a PRISM score < 5 had a survival rate of 100%, whereas patients with an APSC score > 30 and a PRISM score > 25 had a mortality rate of 100%. The area under the receiver operating characteristic (ROC) curve for APSC, PRISM and TISS was 0.847, 0.826 and 0.793, respectively. CONCLUSION: APSC, PRISM and TISS describe accurately severity of illness in infants and children with cardiopulmonary insufficiency after cardiac surgery and all scores identify those patients at increased risk for mortality. PMID- 8227116 TI - Coronary artery aneurysm. AB - A 55 year old man presented with a 2 year history of angina of effort complicated by two myocardial infarctions. Coronary angiography confirmed fusiform aneurysm of the right coronary artery and disease of the left anterior descending coronary artery. The patient underwent surgery with exclusion of the right coronary artery aneurysm and reversed saphenous vein graft, and graft to the left anterior descending coronary artery, with uneventful recovery. The clinical and radiological features are reviewed, and the surgical options considered. PMID- 8227117 TI - Surgical removal of intracardiac foreign bodies. AB - When non-surgical methods for removing intracardiac foreign body have failed, surgical removal should be attempted. Two patients who had dislodged ventriculoatrial shunts for hydrocephalus underwent open removal of catheters, one from the pulmonary artery while the other from the right atrium. Both were successfully retrieved without cardiopulmonary bypass. We describe the clinical and radiological features, with a review of the indications and techniques available. PMID- 8227118 TI - Investigation of nonspecific neurogenic thoracic outlet syndrome. AB - OBJECTIVE: To determine the value of investigations for nonspecific neurogenic thoracic outlet syndrome (NNTOS) and formulate a rational diagnostic approach. DESIGN: Retrospective review. SETTING: Thoracic surgery clinic. PATIENTS: Fifty six patients referred with a presumptive diagnosis of NNTOS: Patients with arterial, venous, or true (classic) neurogenic TOS were excluded. INTERVENTIONS: All patients underwent careful history and examination. Fifty-three had cervical spine radiographs, 45 had electrophysiologic testing, and 30 patients had arterial doppler studies. MAIN OUTCOME MEASURES: The value of investigations that documented a diagnosis other than NNTOS could be assessed but objective evaluation of investigations that appeared to support a diagnosis of NNTOS was not possible. No standard diagnostic criteria exist for NNTOS: RESULTS: Forty (71%) patients were female. Thirty-four (61%) patients had a pulse deficit with provocative maneuvers. Cervical spine radiographs showed cervical ribs in 10 patients and degenerative disease in 11 patients. Electrophysiologic testing demonstrated carpal tunnel syndrome in 11 patients. Arterial Doppler studies were strongly positive for TOS in 10 patients. Thirty-six (64%) patients were diagnosed with conditions other than NNTOS: The diagnosis of NNTOS was accepted in 20 (36%) patients. All 20 patients were initially treated conservatively but 3 (5%) eventually underwent surgery. CONCLUSIONS: NNTOS is a diagnosis of exclusion. Two thirds of patients referred to a thoracic surgery clinic were eventually diagnosed with conditions other than NNTOS: Objective evaluation of investigations that may support a diagnosis of NNTOS is not currently possible. PMID- 8227119 TI - Diagnostic algorithm for acute superior vena caval obstruction (SVCO). AB - Acute SVCO presents a serious diagnostic and therapeutic dilemma to the thoracic surgeon. It is highly desirable to obtain a definitive tissue diagnosis but this requirement must be balanced against the risks inherent in invasive diagnostic procedures in these oft-times critically ill patients. In the past 5 years we have developed an algorithm that has been used successfully in 18 patients presenting with acute SVCO. The decision tree consists of scalene node biopsy, bronchoscopy and mediastinoscopy with categorization into low and high risk groups in the latter. The gender distribution of the 18 patients was 12 male and 6 female with the ages being 58.3 +/- 16.3 and 64.2 +/- 11.2 years respectively. The duration of symptoms range from 1.5 to 12 weeks and averaged 4.5 +/- 3.2 weeks. Two patients had palpable scalene nodes which were positive at biopsy. Bronchoscopy was positive in 5 out of 11 examinations. In 5 instances it was not done. Twelve patients underwent various forms of mediastinal biopsy and one underwent sternotomy. Seven patients requiring mediastinoscopy were judged to be high risk as defined by severe airway and vascular obstruction. Perioperative difficulties occurred in two patients due to cardiorespiratory factors associated with the obstruction. Both patients were undergoing cervical mediastinoscopy under general anesthesia and no difficulties were encountered in high risk patients when local anesthesia was used to perform anterior mediastinotomy. Five patients that were considered low risk all underwent uncomplicated procedures under general anesthesia. Thirteen patients proved to have bronchogenic carcinoma and a specific tissue diagnosis was obtained in 12. The undetermined diagnosis was in a patient who had prior radiotherapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227120 TI - Endobronchial non-Hodgkin's lymphoma. AB - Endobronchial involvement in non-Hodgkin's lymphoma is rare even in the presence of advanced disease. Two cases of endobronchial non-Hodgkin's lymphoma are presented and the findings in 31 previously reported cases reviewed. There were 16 cases with diffuse involvement in the presence of intra or extrathoracic non Hodgkin's lymphoma and 15 cases with central airways involvement in the absence of clinically apparent systemic disease. Chemotherapy with or without radiotherapy is mandatory for all patients with disseminated disease and for those patients with primary endobronchial non-Hodgkin's lymphoma with residual disease after resection, or if resection is not feasible. Older patients should be treated with curative intent unless concomitant intercurrent illness precludes combination chemotherapy. Rapid deterioration in dyspnoea from airway obstruction in non-Hodgkin's lymphoma may be relieved with a self-expanding endobronchial stent. PMID- 8227121 TI - Membrane-associated lamins in Xenopus egg extracts: identification of two vesicle populations. AB - Nuclear lamin isoforms of vertebrates can be divided into two major classes. The B-type lamins are membrane associated throughout the cell cycle, whereas A-type lamins are recovered from mitotic cell homogenates in membrane-free fractions. A feature of oogenesis in birds and mammals is the nearly exclusive presence of B type lamins in oocyte nuclear envelopes. In contrast, oocytes and early cleavage embryos of the amphibian Xenopus laevis are believed to contain a single lamin isoform, lamin LIII, which after nuclear envelope breakdown during meiotic maturation is reported to be completely soluble. Consequently, we have reexamined the lamin complement of Xenopus oocyte nuclear envelopes, egg extracts, and early embryos. An mAb (X223) specific for the homologous B-type lamins B2 of mouse and LII of Xenopus somatic cells (Hoger, T., K. Zatloukal, I. Waizenegger, and G. Krohne. 1990. Chromosoma. 99:379-390) recognized a Xenopus oocyte nuclear envelope protein biochemically distinct from lamin LIII and very similar or identical to somatic cell lamin LII. Oocyte lamin LII was detectable in nuclear envelopes of early cleavage embryos. Immunoblotting of fractionated egg extracts revealed that approximately 20-23% of lamin LII and 5-7% of lamin LIII were membrane associated. EM immunolocalization demonstrated that membrane-bound lamins LII and LIII are associated with separate vesicle populations. These findings are relevant to the interpretation of nuclear reconstitution experiments using Xenopus egg extracts. PMID- 8227122 TI - The cell proliferation-associated antigen of antibody Ki-67: a very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins. AB - The antigen defined by mAb Ki-67 is a human nuclear protein the expression of which is strictly associated with cell proliferation and which is widely used in routine pathology as a "proliferation marker" to measure the growth fraction of cells in human tumors. Ki-67 detects a double band with apparent molecular weights of 395 and 345 kD in immunoblots of proteins from proliferating cells. We cloned and sequenced the full length cDNA, identified two differentially spliced isoforms of mRNA with open reading frames of 9,768 and 8,688 bp encoding for this cell proliferation-associated protein with calculated molecular weights of 358,761 D and 319,508 D, respectively. New mAbs against a bacterially expressed part and a synthetic polypeptide deduced from the isolated cDNA react with the native Ki-67 antigen, thus providing a circle of evidence that we have cloned the authentic Ki-67 antigen cDNA. The central part of the Ki-67 antigen cDNA contains a large 6,845-bp exon with 16 tandemly repeated 366-bp elements, the "Ki-67 repeats", each including a highly conserved new motif of 66 bp, the "Ki-67 motif", which encodes for the epitope detected by Ki-67. Computer analysis of the nucleic acid and the deduced amino acid sequence of the Ki-67 antigen confirmed that the cDNA encodes for a nuclear and short-lived protein without any significant homology to known sequences. Ki-67 antigen-specific antisense oligonucleotides inhibit the proliferation of IM-9 cell line cells, indicating that the Ki-67 antigen may be an absolute requirement for maintaining cell proliferation. We conclude that the Ki-67 antigen defines a new category of cell cycle-associated nuclear nonhistone proteins. PMID- 8227123 TI - Dissociation of the dorsal-cactus complex and phosphorylation of the dorsal protein correlate with the nuclear localization of dorsal. AB - The formation of dorsal-ventral polarity in Drosophila requires the asymmetric nuclear localization of the dorsal protein along the D/V axis. This process is regulated by the action of the dorsal group genes and cactus. We show that dorsal and cactus are both phosphoproteins that form a stable cytoplasmic complex, and that the cactus protein is stabilized by its interaction with dorsal. The dorsal cactus complex dissociates when dorsal is targeted to the nucleus. While the phosphorylation of cactus remains apparently unchanged during early embryogenesis, the phosphorylation state of dorsal correlates with its release from cactus and with its nuclear localization. This differential phosphorylation event is regulated by the dorsal group pathway. PMID- 8227124 TI - Cloning and characterization of PAS5: a gene required for peroxisome biogenesis in the methylotrophic yeast Pichia pastoris. AB - The biogenesis and maintenance of cellular organelles is of fundamental importance in all eukaryotic cells. One such organelle is the peroxisome. The establishment of a genetic system to study peroxisome biogenesis in the methylotrophic yeast Pichia pastoris has yielded many different complementation groups of peroxisomal assembly (pas) or peroxisome-deficient (per) mutants. Each appears to be deficient in functional peroxisomes. One of these mutants, pas5, has been characterized, complemented, and the gene sequenced. Ultrastructural studies show that normal peroxisomes are not present in pas5, but aberrant peroxisomal structures resembling "membranous ghosts" are frequently observed. The "peroxisome ghosts" appear to be induced and segregated to daughter cells normally. Biochemical fractionation analysis of organelles of the pas5 mutant reveals that peroxisomal matrix enzymes are induced normally but are found mostly in the cytosol. However, purification of peroxisome ghosts from the mutant shows that small amounts (< 5%) of matrix enzymes are imported. The PAS5 gene was cloned and found to encode a 127-kD protein, which contains a 200-amino acid-long region of homology with PAS1, NEM-sensitive factor (NSF), and other related ATPases. Weak homology to a yeast myosin was also observed. The gene is not essential for growth on glucose but is essential for growth on oleic acid and methanol. The role of PAS5 in peroxisome biogenesis is discussed. PMID- 8227125 TI - The v-sis protein retains biological activity as a type II membrane protein when anchored by various signal-anchor domains, including the hydrophobic domain of the bovine papilloma virus E5 oncoprotein. AB - Membrane-anchored forms of the v-sis oncoprotein have been previously described which are oriented as type I transmembrane proteins and which efficiently induce autocrine transformation. Several examples of naturally occurring membrane anchored growth factors have been identified, but all exhibit a type I orientation. In this work, we wished to construct and characterize membrane anchored growth factors with a type II orientation. These experiments were designed to determine whether type II membrane-anchored growth factors would in fact exhibit biological activity. Additionally, we wished to determine whether the hydrophobic domain of the E5 oncoprotein of bovine papilloma virus (BPV) can function as a signal-anchor domain to direct type II membrane insertion. Type II derivatives of the v-sis oncoprotein were constructed, with the NH2 terminus intracellular and the COOH terminus extracellular, by substituting the NH2 terminal signal sequence with the signal-anchor domain of a known type II membrane protein. The signal-anchor domains of neuraminidase (NA), asialoglycoprotein receptor (ASGPR) and transferrin receptor (TR) all yielded biologically active type II derivatives of the v-sis oncoprotein. Although transforming all of the type II signal/anchor-sis proteins exhibited a very short half-life. The short half-life exhibited by the signal/anchor-sis constructs suggests that, in some cases, cellular transformation may result from the synthesis of growth factors so labile that they activate undetectable autocrine loops. The E5 oncoprotein encoded by BPV exhibits amino acid sequence similarity with PDGF, activates the PDGF beta-receptor, and thus resembles a miniature membrane-anchored growth factor with a putative type II orientation. The hydrophobic domain of the E5 oncoprotein, when substituted in place of the signal sequence of v-sis, was indistinguishable compared with the signal-anchor domains of NA, TR, and ASGPR, demonstrating its ability to function as a signal-anchor domain. NIH 3T3 cells transformed by the signal/anchor-sis constructs exhibited morphological reversion upon treatment with suramin, indicating a requirement for ligand/receptor interactions in a suramin-sensitive compartment, most likely the cell surface. In contrast, NIH 3T3 cells transformed by the E5 oncoprotein did not exhibit morphological reversion in response to suramin. PMID- 8227126 TI - Biochemical dissection of AP-1 recruitment onto Golgi membranes. AB - Recruitment of the Golgi-specific AP-1 adaptor complex onto Golgi membranes is thought to be a prerequisite for clathrin coat assembly on the TGN. We have used an in vitro assay to examine the translocation of cytosolic AP-1 onto purified Golgi membranes. Association of AP-1 with the membranes required GTP or GTP analogues and was inhibited by the fungal metabolite, brefeldin A. In the presence of GTP gamma S, binding of AP-1 to Golgi membranes was strictly dependent on the concentration of cytosol added to the assay. AP-1 recruitment was also found to be temperature dependent, and relatively rapid at 37 degrees C, following a lag period of 3 to 4 min. Using only an adaptor-enriched fraction from cytosol, purified myristoylated ARF1, and Golgi membranes, the GTP gamma S dependent recruitment of AP-1 could be reconstituted. Our results show that the association of the AP-1 complex with Golgi membranes, like the coatomer complex, requires ARF, which accounts for the sensitivity of both to brefeldin A. In addition, they provide the basis for a model for the early biochemical events that lead to clathrin-coated vesicle formation on the TGN. PMID- 8227127 TI - Degranulation of individual mast cells in response to Ca2+ and guanine nucleotides: an all-or-none event. AB - Widespread experience indicates that application of suboptimal concentrations of stimulating ligands (secretagogues) to secretory cells elicits submaximal extents of secretion. Similarly, for permeabilized secretory cells, the extent of secretion is related to the concentration of applied intracellular effectors. We investigated the relationship between the extent of secretion from mast cells (assessed as the release of hexosaminidase) and the degranulation (exocytosis) responses of individual cells. For permeabilized mast cells stimulated by the effector combination Ca2+ plus GTP-gamma-S and for intact cells stimulated by the Ca2+ ionophore ionomycin, we found that exocytosis has the characteristics of an all-or-none process at the level of the individual cells. With a suboptimal stimulus, the population comprised only totally degranulated cells and fully replete cells. In contrast, a suboptimal concentration of compound 48/80 applied to intact cells induced a partial degree of degranulation. This was determined by observing the morphological changes accompanying degranulation by light and electron microscopy and also as a reduction in the intensity of light scattered at 90 degrees, indicative of a change in the cell-refractive index. These results may be explained by the existence of a threshold sensitivity to the combined effectors that is set at the level of individual cells and not at the granule level. We used flow cytometry to establish the relationship between the extent of degranulation in individual rat peritoneal mast cells and the extent of secretion in the population (measured as the percentage release of total hexosaminidase). For comparison, secretion was also elicited by applying the Ca2+ ionophore ionomycin or compound 48/80 to intact cells. For permeabilized cells and also for intact cells stimulated with the ionophore, levels of stimulation that generate partial secretion gave rise to bimodal frequency distributions of 90 degrees light scatter. In contrast, a partial stimulus to secretion by compound 48/80 resulted in a single population of partially degranulated cells, the degree of degranulation varying across the cell population. The difference between the all or-none responses of the permeabilized or ionophore-treated cells and the graded responses of cells activated by compound 48/80 is likely to stem from differences in the effective calcium stimulus. Whereas cell stimulated with receptor-directed agonists can undergo transient and localized Ca2+ changes, a homogeneous and persistent stimulus is sensed at every potential exocytotic site in the permeabilized cells. PMID- 8227128 TI - Caveolin forms a hetero-oligomeric protein complex that interacts with an apical GPI-linked protein: implications for the biogenesis of caveolae. AB - Glycosyl-phosphatidylinositol (GPI)-linked proteins are transported to the apical surface of epithelial cells where they undergo cholesterol-dependent clustering in membrane micro-invaginations, termed caveolae or plasmalemmal vesicles. However, the sorting machinery responsible for this caveolar-clustering mechanism remains unknown. Using transfected MDCK cells as a model system, we have identified a complex of cell surface molecules (80, 50, 40, 22-24, and 14 kD) that interact in a pH- and cholesterol-dependent fashion with an apical recombinant GPI-linked protein. A major component of this hetero-oligomeric protein complex is caveolin, a type II transmembrane protein. As this hetero oligomeric caveolin complex is detectable almost immediately after caveolin synthesis, our results suggest that caveolae may assemble intracellularly during transport to the cell surface. As such, our studies have implications for understanding both the intracellular biogenesis of caveolae and their subsequent interactions with GPI-linked proteins in epithelia and other cell types. PMID- 8227129 TI - The major myosin-binding domain of skeletal muscle MyBP-C (C protein) resides in the COOH-terminal, immunoglobulin C2 motif. AB - A common feature shared by myosin-binding proteins from a wide variety of species is the presence of a variable number of related internal motifs homologous to either the Ig C2 or the fibronectin (Fn) type III repeats. Despite interest in the potential function of these motifs, no group has clearly demonstrated a function for these sequences in muscle, either intra- or extracellularly. We have completed the nucleotide sequence of the fast type isoform of MyBP-C (C protein) from chicken skeletal muscle. The deduced amino acid sequence reveals seven Ig C2 sets and three Fn type III motifs in MyBP-C. alpha-chymotryptic digestion of purified MyBP-C gives rise to four peptides. NH2-terminal sequencing of these peptides allowed us to map the position of each along the primary structure of the protein. The 28-kD peptide contains the NH2-terminal sequence of MyBP-C, including the first C2 repeat. It is followed by two internal peptides, one of 5 kD containing exclusively spacer sequences between the first and second C2 motifs, and a 95-kD fragment containing five C2 domains and three fibronectin type III motifs. The C-terminal sequence of MyBP-C is present in a 14-kD peptide which contains only the last C2 repeat. We examined the binding properties of these fragments to reconstituted (synthetic) myosin filaments. Only the COOH terminal 14-kD peptide is capable of binding myosin with high affinity. The NH2 terminal 28-kD fragment has no myosin-binding, while the long internal 100-kD peptide shows very weak binding to myosin. We have expressed and purified the 14 kD peptide in Escherichia coli. The recombinant protein exhibits saturable binding to myosin with an affinity comparable to that of the 14-kD fragment obtained by proteolytic digestion (1/2 max binding at approximately 0.5 microM). These results indicate that the binding to myosin filaments is mainly restricted to the last 102 amino acids of MyBP-C. The remainder of the molecule (1,032 amino acids) could interact with titin, MyBP-H (H protein) or thin filament components. A comparison of the highly conserved Ig C2 domains present at the COOH-terminus of five MyBPs thus far sequenced (human slow and fast MyBP-C, human and chicken MyBP-H, and chicken MyBP-C) was used to identify residues unique to these myosin binding Ig C2 repeats. PMID- 8227130 TI - Non-sarcomeric mode of myosin II organization in the fibroblast lamellum. AB - The organization of myosin in the fibroblast lamellum was studied by correlative fluorescence and electron microscopy after a novel procedure to reveal its underlying morphology. An X-rhodamine analog of conventional smooth muscle myosin (myosin II) that colocalized after microinjection with endogenous myosin was used to trace myosin distribution in living fibroblasts. Then, the same cells were examined by EM of platinum replicas. To visualize the structural arrangement of myosin, other cytoskeletal fibrillar structures had to be removed: microtubules were depolymerized by nocodazole treatment of the living cells before injection of myosin; continued nocodazole treatment also induced the intermediate filaments to concentrate near the nucleus, thus removing them from the lamellar region; actin filaments were removed after lysis of the cells by incubation of the cytoskeletons with recombinant gelsolin. Possible changes in myosin organization caused by this treatment were examined by fluorescence microscopy. No significant differences in myosin distribution patterns between nocodazole-treated and control cells were observed. Cell lysis and depletion of actin also did not induce reorganization of myosin as was shown by direct comparison of myosin distribution in the same cells in the living state and after gelsolin treatment. EM of the well-spread, peripheral regions of actin-depleted cytoskeletons revealed a network of bipolar myosin mini-filaments, contracting each other at their terminal, globular regions. The morphology of this network corresponded well to the myosin distribution observed by fluorescence microscopy. A novel mechanism of cell contraction by folding of the myosin filament network is proposed. PMID- 8227131 TI - The kinesin-like protein KLP61F is essential for mitosis in Drosophila. AB - We report here that disruption of a recently discovered kinesin-like protein in Drosophila melanogaster, KLP61F, results in a mitotic mutation lethal to the organism. We show that in the absence of KLP61F function, spindle poles fail to separate, resulting in the formation of monopolar mitotic spindles. The resulting phenotype of metaphase arrest with polyploid cells is reminiscent of that seen in the fungal bimC and cut7 mutations, where it has also been shown that spindle pole bodies are not segregated. KLP61F is specifically expressed in proliferating tissues during embryonic and larval development, consistent with a primary role in cell division. The structural and functional homology of the KLP61F, bimC, cut7, and Eg5 kinesin-like proteins demonstrates the existence of a conserved family of kinesin-like molecules important for spindle pole separation and mitotic spindle dynamics. PMID- 8227132 TI - Roles of kinesin and kinesin-like proteins in sea urchin embryonic cell division: evaluation using antibody microinjection. AB - Previous studies suggest that kinesin heavy chain (KHC) is associated with ER derived membranes that accumulate in the mitotic apparatus in cells of early sea urchin embryos (Wright, B. D., J. H. Henson, K. P. Wedaman, P. J. Willy, J. N. Morand, and J. M. Scholey. 1991. J. Cell Biol. 113:817-833). Here, we report that the microinjection of KHC-specific antibodies into these cells has no effect on mitosis or ER membrane organization, even though one such antibody, SUK4, blocks kinesin-driven motility in vitro and in mammalian cells. Microinjected SUK4 was localized to early mitotic figures, suggesting that it is able to access kinesin in spindles. In contrast to KHC-specific antibodies, two antibodies that react with kinesin-like proteins (KLPs), namely CHO1 and HD, disrupted mitosis and prevented subsequent cell division. CHO1 is thought to exert this effect by blocking the activity of a 110-kD KLP. The relevant target of HD, which was raised against the KHC motor domain, is unknown; HD may disrupt mitosis by interfering with an essential spindle KLP but not with KHC itself, as preabsorption of HD with KHC did not alter its ability to block mitosis. These data indicate that some KLPs have essential mitotic functions in early sea urchin embryos but KHC itself does not. PMID- 8227133 TI - Differential regulation of multiple gap junction transcripts and proteins during rat liver regeneration. AB - The mRNA and protein expression of alpha 1 (connexin 43), beta 1 (connexin 32), and beta 2 (connexin 26) gap junction genes were examined in the regenerating rat liver after 70% partial hepatectomy (PH). Expression of beta 1 and beta 2 steady state mRNA levels changed minimally until 12 h after PH when both transcripts decreased to approximately 15% of baseline values. A similar decrease in assembled connexin levels was detected by immunoblot and indirect immunofluorescence at 18 h after PH. Both transcripts simultaneously increased between 24 and 42 h and again rapidly decreased by 48 h post-PH. beta 1 and beta 2 assembled gap junction protein expression increased at 48 h post-PH and rapidly decreased by 56 h. By 72 to 84 h post-PH, beta 1 and beta 2 mRNA and assembled protein expression returned to near baseline levels and were maintained. Interestingly, inhibition of protein synthesis with cycloheximide completely inhibited disappearance of the beta 2 transcript, in contrast to beta 1 mRNA which was unaffected. Nuclear run-on assays showed no change in transcriptional rates for either gene during the regenerative period. However, both beta 1 and beta 2 transcripts exhibited significantly decreased mRNA half-lives at 12 h post PH (3.8 and 3.7 h, respectively) relative to those at 0 h (10.9 and 6.1 h, respectively). Surprisingly, although the transcriptional rate for alpha 1 was similar to that observed for beta 2, no alpha 1 transcripts were detectable by northern or RNase protection analysis. The results suggest that in the regenerating rat liver, beta 1 and beta 2 gap junction genes are not regulated at the transcriptional level. Rather, the cyclical modulation of their steady-state transcripts is regulated primarily by posttranscriptional events of which mRNA stability is at least one critical factor in the control process. PMID- 8227134 TI - Clustering of the acetylcholine receptor by the 43-kD protein: involvement of the zinc finger domain. AB - A postsynaptic membrane-associated protein of M(r) 43,000 (43-kD protein) is involved in clustering of the nicotinic acetylcholine receptor (AChR) at the neuromuscular junction. Previous studies have shown that recombinant mouse 43-kD protein forms membrane-associated clusters when expressed in Xenopus oocytes. Coexpression with the AChR results in colocalization of the receptor with the 43 kD protein clusters (Froehner, S. C., C. W. Luetje, P. B. Scotland, and J. Patrick, 1990. Neuron. 5:403-410). To understand the mechanism of this clustering, we have studied the role of the carboxy-terminal region of the 43-kD protein. The amino acid sequence of this region predicts two tandem zinc finger structures followed by a serine phosphorylation site. Both Torpedo 43-kD protein and the carboxy-terminal region of the mouse 43-kD protein bind radioisotopic zinc. Mutation of two histidine residues in this predicted domain greatly attenuates zinc binding, lending support to the proposal that this region forms zinc fingers. When expressed in oocytes, the ability of this mutant 43-kD protein to form clusters is greatly reduced. Its ability to interact with AChR, however, is retained. In contrast, a mutation that eliminates the potential serine phosphorylation site has no effect on clustering of the 43-kD protein or on interaction with the AChR. These findings suggest that protein interactions via the zinc finger domain of the 43-kD protein may be important for AChR clustering at the synapse. PMID- 8227135 TI - Clustering and immobilization of acetylcholine receptors by the 43-kD protein: a possible role for dystrophin-related protein. AB - Recombinant acetylcholine receptors (AChRs) expressed on the surface of cultured fibroblasts become organized into discrete membrane domains when the 43-kD postsynaptic protein (43k) is co-expressed in the same cells (Froehner, S.C., C. W. Luetje, P. B. Scotland, and J. Patrick, 1990. Neuron. 5:403-410; Phillips, W. D., M. C. Kopta, P. Blount, P. D. Gardner, J. H. Steinbach, and J. P. Merlie. 1991. Science (Wash. DC). 251:568-570). Here we show that AChRs present on the fibroblast cell surface prior to transfection of 43k are recruited into 43k-rich membrane domains. Aggregated AChRs show increased resistance to extraction with Triton X-100, suggesting a 43k-dependent linkage to the cytoskeleton. Myotubes of the mouse cell line C2 spontaneously display occasional AChR/43k-rich membrane domains that ranged in diameter up to 15 microns, but expressed many more when 43k was overexpressed following transfection of 43k cDNA. However, the membrane domains induced by recombinant 43k were predominantly small (< or = 2 microns). We were then interested in whether the cytoskeletal component, dystrophin related protein (DRP; Tinsley, J. M., D. J. Blake, A. Roche, U. Fairbrother, J. Riss, B. C. Byth, A. E. Knight, J. Kendrick-Jones, G. K. Suthers, D. R. Love, Y. H. Edwards, and K. E. Davis, 1992. Nature (Lond.). 360:591-593) contributed to the development of AChR clusters. Immunofluorescent anti-DRP staining was present at the earliest stages of AChR clustering at the neuromuscular synapse in mouse embryos and was also concentrated at the large AChR-rich domains on nontransfected C2 myotubes. Surprisingly, anti-DRP staining was concentrated mainly at the large, but not the small AChR clusters on C2 myotubes suggesting that DRP may be principally involved in permitting the growth of AChR clusters. PMID- 8227136 TI - Mechanical strain induces growth of vascular smooth muscle cells via autocrine action of PDGF. AB - The effect of cyclic mechanical strain on growth of neonatal rat vascular smooth muscle (VSM) cells were examined. Cells were grown on silicone elastomer plates subjected to cyclic strain (60 cycle/min) by application of a vacuum under the plates. A 48 h exposure to mechanical strain increased the basal rate of thymidine incorporation by threefold and increased cell number by 40% compared with cells grown on stationary rubber plates. Strain also increased the rate of thymidine incorporation in response to alpha-thrombin (from 15- to 33-fold), but not to PDGF. As determined by thymidine autoradiography, strain alone induced a fourfold increase in labeled nuclei at the periphery of dishes, where strain is maximal, and a 2-3-fold increase at the center of dishes. Strain appeared to induce the production of an autocrine growth factor(s), since conditioned medium from cells subjected to strain induced a fourfold increase in DNA synthesis in control cells. Western blots of medium conditioned on the cells subjected to strain indicate that the cells secrete both AA and BB forms of PDGF in response to strain. Northern blots of total cell RNA from cells exposed to strain for 24 h show increased steady-state level of mRNA for PDGF-A. Lastly, polyclonal antibodies to the AA form of PDGF reduced by 75% the mitogenic effect of strain and polyclonal antibodies to AB-PDGF reduced mitogenicity by 50%. Antibodies to bFGF did not significantly reduce the strain-induced thymidine incorporation. Thus, the mechanism of strain-induced growth appears to involve the intermediary action of secreted PDGF. PMID- 8227137 TI - A heparin-binding activity on Leishmania amastigotes which mediates adhesion to cellular proteoglycans. AB - The intracellular amastigote form of leishmania is responsible for the cell-to cell spread of leishmania infection in the mammalian host. In this report, we identify a high-affinity, heparin-binding activity on the surface of the amastigote form of leishmania. Amastigotes of Leishmania amazonensis bound approximately 120,000 molecules of heparin per cell, with a Kd of 8.8 x 10(-8) M. This heparin-binding activity mediates the adhesion of amastigotes to mammalian cells via heparan sulfate proteoglycans, which are expressed on the surface of mammalian cells. Amastigotes bound efficiently to a variety of adherent cells which express cell-surface proteoglycans. Unlike wild-type CHO cells, which bound amastigotes avidly, CHO cells with genetic deficiencies in heparan sulfate proteoglycan biosynthesis or cells treated with heparitinase failed to bind amastigotes even at high parasite-input dosages. Cells which express normal levels of undersulfated heparan bound amastigotes nearly as efficiently as did wild-type cells. The adhesion of amastigotes to wild-type nonmyeloid cells was almost completely inhibited by the addition of micromolar amounts of soluble heparin or heparan sulfate but not by the addition of other sulfated polysaccharides.l Binding of amastigotes to macrophages, however, was inhibited by only 60% after pretreatment of amastigotes with heparin, suggesting that macrophages have an additional mechanism for recognizing amastigotes. These results suggest that leishmania amastigotes express a high-affinity, heparin binding activity on their surface which can interact with heparan sulfate proteoglycans on mammalian cells. This interaction may represent an important first step in the invasion of host cells by amastigotes. PMID- 8227138 TI - Regulation of alpha 6 beta 1 integrin laminin receptor function by the cytoplasmic domain of the alpha 6 subunit. AB - The alpha 6 beta 1 integrin is expressed on the macrophage surface in an inactive state and requires cellular activation with PMA or cytokines to function as a laminin receptor (Shaw, L. M., J. M. Messier, and A. M. Mercurio. 1990. J. Cell Biol. 110:2167-2174). In the present study, the role of the alpha 6 subunit cytoplasmic domain in alpha 6 beta 1 integrin activation was examined. The use of P388D1 cells, an alpha 6-integrin deficient macrophage cell line, facilitated this analysis because expression of either the alpha 6A or alpha 6B subunit cDNAs restores their activation responsive laminin adhesion (Shaw, L. S., M. Lotz, and A. M. Mercurio. 1993. J. Biol. Chem. 268:11401-11408). A truncated alpha 6 cDNA, alpha 6-delta CYT, was constructed in which the human cytoplasmic domain sequence was deleted after the GFFKR pentapeptide. Expression of this cDNA in P388D1 cells resulted in the surface expression of a chimeric alpha 6-delta CYT beta 1 integrin that was unable to mediate laminin adhesion or increase this adhesion in response to PMA under normal conditions, i.e., in medium that contained physiological concentrations of Ca++ and Mg++. The alpha 6A-delta CYT transfectants adhered to laminin, however, when Ca++/Mg++ was replaced with 150 microM Mn++. We also assessed the role of serine phosphorylation in the regulation of alpha 6A beta 1 integrin function by site-directed mutagenesis of the two serine residues present in the alpha 6A cytoplasmic domain because this domain is phosphorylated on serine residues in response to stimuli that activate the laminin receptor function of alpha 6 A beta 1. Point mutations were introduced in the alpha 6A cDNA that changed either serine residue #1064 (M1) or serine residue #1071 (M2) to alanine residues. In addition, a double mutant (M3) was constructed in which both serine residues were changed to alanine residues. P388D1 transfectants which expressed these serine mutations adhered to laminin in response to PMA to the same extent as cells transfected with wild-type alpha 6A cDNA. These findings provide evidence for a novel mode of integrin regulation that is distinct from that reported for other regulated integrins (O'Toole, T. E., D. Mandelman, J. Forsyth, S. J. Shattil, E. F. Plow, and M. H. Ginsberg. 1991. Science (Wash. DC). 254:845-847. Hibbs, M. L., H. Xu, S. A. Stacker, and T. A. Springer. 1991. Science (Wash. DC). 251:1611-1613), and they demonstrate that serine phosphorylation of the alpha 6A cytoplasmic domain is not involved in this regulation. PMID- 8227139 TI - Isolation of the yeast nuclear pore complex. AB - Nuclear pore complexes (NPCs) have been isolated from the yeast Saccharomyces. Negative stain electron microscopy of the isolated NPCs and subsequent image reconstruction revealed the octagonal symmetry and many of the ultrastructural features characteristic of vertebrate NPCs. The overall dimensions of the yeast NPC, both in its isolated form as well as in situ, are smaller than its vertebrate counterpart. However, the diameter of the central structures are similar. The isolated yeast NPC has a sedimentation coefficient of approximately 310 S and an M(r) of approximately 66 MD. It retains all but one of the eight known NPC proteins. In addition it contains as many as 80 uncharacterized proteins that are candidate NPC proteins. PMID- 8227140 TI - Reconstitution of nuclear protein transport with semi-intact yeast cells. AB - We have developed an in vitro nuclear protein import reaction from semi-intact yeast cells. The reaction uses cells that have been permeabilized by freeze-thaw after spheroplast formation. Electron microscopic analysis and antibody-binding experiments show that the nuclear envelope remains intact but the plasma membrane is perforated. In the presence of ATP and cytosol derived from yeast or mammalian cells, a protein containing the nuclear localization sequence (NLS) of SV40 large T-antigen is transported into the nucleus. Proteins with mutant NLSs are not imported. In the absence of cytosol, binding of NLS-containing proteins occurs at the nuclear envelope. N-ethylmaleimide treatment of the cytosol as well as antibodies to the nuclear pore protein Nsp1 inhibit import but not binding to the nuclear envelope. Yeast mutants defective in nuclear protein transport were tested in the in vitro import reaction. Semi-intact cells from temperature sensitive nsp1 mutants failed to import but some binding to the nuclear envelope was observed. On the other hand, no binding and thus no import into nuclei was observed in semi-intact nsp49 cells which are mutated in another nuclear pore protein. Np13 mutants, which are defective for nuclear protein import in vivo, were also deficient in the binding step under the in vitro conditions. Thus, the transport defect in these mutants is at the level of the nucleus and the point at which nuclear transport is blocked can be defined. PMID- 8227141 TI - GTP hydrolysis by complexes of the signal recognition particle and the signal recognition particle receptor. AB - Translocation of proteins across the endoplasmic reticulum membrane is a GTP dependent process. The signal recognition particle (SRP) and the SRP receptor both contain subunits with GTP binding domains. One GTP-dependent reaction during protein translocation is the SRP receptor-mediated dissociation of SRP from the signal sequence of a nascent polypeptide. Here, we have assayed the SRP and the SRP receptor for GTP binding and hydrolysis activities. GTP hydrolysis by SRP was not detected, so the maximal GTP hydrolysis rate for SRP was estimated to be < 0.002 mol GTP hydrolyzed x mol of SRP-1 x min-1. The intrinsic GTP hydrolysis activity of the SRP receptor ranged between 0.02 and 0.04 mol GTP hydrolyzed x mol of SRP receptor-1 x min-1. A 40-fold enhancement of GTP hydrolysis activity relative to that observed for the SRP receptor alone was obtained when complexes were formed between SRP and the SRP receptor. GTP hydrolysis activity was inhibited by GDP, but not by ATP. Extended incubation of the SRP or the SRP receptor with GTP resulted in substoichiometric quantities of protein-bound ribonucleotide. SRP-SRP receptor complexes engaged in GTP hydrolysis were found to contain a minimum of one bound guanine ribonucleotide per SRP-SRP receptor complex. We conclude that the GTP hydrolysis activity described here is indicative of one of the GTPase cycles that occur during protein translocation across the endoplasmic reticulum. PMID- 8227142 TI - The cytoplasmic droplet of rat epididymal spermatozoa contains saccular elements with Golgi characteristics. AB - The cytoplasmic droplet of epididymal spermatozoa is a small localized outpouching of cytoplasm of the tail of unknown significance. EM revealed flattened saccular elements as the near exclusive membranous component of the droplet. Light and electron microscopic immunolabeling for Golgi/TGN markers showed these saccules to be reactive for antibodies to TGN38, protein affinity purified alpha 2,6 sialyltransferase, and anti-human beta 1,4 galactosyltransferase. The saccules were isolated by subcellular fractionation and antibodies raised against this fraction immunolabeled the saccules of the droplet in situ as well as the Golgi region of somatic epithelial cells lining the epididymis. The isolated droplet fraction was enriched in galactosyltransferase and sialyltransferase activities, and endogenous glycosylation assays identified the modification of several endogenous glycopeptides. EM lectin staining in situ demonstrated galactose and N-acetyl galactosamine constituents in the saccules. Endocytic studies with cationic and anionic ferritin as well as HRP failed to identify the saccules as components of the endocytic apparatus. Epididymal spermatozoa were devoid of markers for the ER as well as the Golgi-associated coatamer protein beta-COP. It is therefore unlikely that the saccular elements of the droplet participate in vesicular protein transport. However, the identification of Golgi/TGN glycosylating activities in the saccules may be related to plasma membrane modifications which occur during epididymal sperm maturation. PMID- 8227143 TI - Type 2X-myosin heavy chain is coded by a muscle fiber type-specific and developmentally regulated gene. AB - We have previously reported the identification of a distinct myosin heavy chain (MyHC) isoform in a major subpopulation of rat skeletal muscle fibers, referred to as 2X fibers (Schiaffino, S., L. Gorza, S. Sartore, L. Saggin, M. Vianello, K. Gundersen, and T. Lomo. 1989. J. Muscle Res. Cell Motil. 10:197-205). However, it was not known whether 2X-MyHC is the product of posttranslational modification of other MyHCs or is coded by a distinct mRNA. We report here the isolation and characterization of cDNAs coding a MyHC isoform that is expressed in type 2X skeletal muscle fibers. 2X-MyHC transcripts differ from other MyHC transcripts in their restriction map and 3' end sequence and are thus derived from a distinct gene. In situ hybridization analyses show that 2X-MyHC transcripts are expressed at high levels in the diaphragm and fast hindlimb muscles and can be coexpressed either with 2B- or 2A-MyHC transcripts in a number of fibers. At the single fiber level the distribution of each MyHC mRNA closely matches that of the corresponding protein, determined by specific antibodies on serial sections. In hindlimb muscles 2X-, 2A-, and 2B-MyHC transcripts are first detected by postnatal day 2-5 and display from the earliest stages a distinct pattern of distribution in different muscles and different fibers. The emergence of type 2 MyHC isoforms thus defines a distinct neonatal phase of fiber type differentiation during muscle development. The functional significance of MyHC isoforms is discussed with particular reference to the velocity of shortening of skeletal muscle fibers. PMID- 8227144 TI - Orientation and three-dimensional organization of actin filaments in dividing cultured cells. AB - The current hypothesis of cytokinesis suggests that contractile forces in the cleavage furrow are generated by a circumferential band of actin filaments. However, relatively little is known about the global organization of actin filaments in dividing cells. To approach this problem we have used fluorescence detected linear dichroism (FDLD) microscopy to measure filament orientation, and digital optical sectioning microscopy to perform three-dimensional reconstructions of dividing NRK cells stained with rhodamine-phalloidin. During metaphase, actin filaments in the equatorial region show a slight orientation along the spindle axis, while those in adjacent regions appear to be randomly distributed. Upon anaphase onset and through cytokinesis, the filaments become oriented along the equator in the furrow region, and along the spindle axis in adjacent regions. The degree of orientation appears to be dependent on cell-cell and cell-substrate adhesions. By performing digital optical sectioning microscopy on a highly spread NRK subclone, we show that actin filaments organize as a largely isotropic cortical meshwork in metaphase cells and convert into an anisotropic network shortly after anaphase onset, becoming more organized as cytokinesis proceeds. The conversion is most dramatic on the adhering ventral surface which shows little or no cleavage activity, and results in the formation of large bundles along the equator. On the dorsal surface, where cleavage occurs actively, actin filaments remain isotropic, showing only subtle alignment late in cytokinesis. In addition, stereo imaging has led to the discovery of a novel set of filaments that are associated with the cortex and traverse through the cytoplasm. Together, these studies provide important insights into the process of actin remodeling during cell division and point to possible additional mechanisms for force generation. PMID- 8227145 TI - Cytoplasmic dynein plays a role in mammalian mitotic spindle formation. AB - The formation and functioning of a mitotic spindle depends not only on the assembly/disassembly of microtubules but also on the action of motor enzymes. Cytoplasmic dynein has been localized to spindles, but whether or how it functions in mitotic processes is not yet known. We have cloned and expressed DNA fragments that encode the putative ATP-hydrolytic sites of the cytoplasmic dynein heavy chain from HeLa cells and from Dictyostelium. Monospecific antibodies have been raised to the resulting polypeptides, and these inhibit dynein motor activity in vitro. Their injection into mitotic mammalian cells blocks the formation of spindles in prophase or during recovery from nocodazole treatment at later stages of mitosis. Cells become arrested with unseparated centrosomes and form monopolar spindles. The injected antibodies have no detectable effect on chromosome attachment to a bipolar spindle or on motions during anaphase. These data suggest that cytoplasmic dynein plays a unique and important role in the initial events of bipolar spindle formation, while any later roles that it may play are redundant. Possible mechanisms of dynein's involvement in mitosis are discussed. PMID- 8227146 TI - Deflagellation of Chlamydomonas reinhardtii follows a rapid transitory accumulation of inositol 1,4,5-trisphosphate and requires Ca2+ entry. AB - C. reinhardtii sheds its flagella in response to acidification. Previously, we showed correlations between pH shock, deflagellation, and inositol 1,4,5 trisphosphate [Ins(1,4,5)P3] production, but 100% of cells deflagellated by 5 s, which was the earliest that Ins(1,4,5)P3 accumulation could be accurately measured by techniques available to us at that time (Quarmby, L. M., Y. G. Yueh, J. L. Cheshire, L. R. Keller, W. J. Snell, and R. C. Crain. J. Cell Biol. 1992. 116:737-744). To learn about the causal relationship between Ins(1,4,5)P3 accumulation and deflagellation, we extended these studies to early times using a continuous-flow rapid-quench device. Within 1 s of acidification to pH 4.3-4.5, 100% of cells deflagellated. A transient peak of Ins(1,4,5)P3 was observed 250 350 ms after pH shock, preceding deflagellation. Preincubation with 10 microM neomycin, which prevents hydrolysis of phosphatidylinositol 4,5-bisphosphate, inhibited both the transient production of Ins(1,4,5)P3 and the subsequent deflagellation. The nonspecific Ca2+ channel blockers La3+ and Cd2+ prevented flagellar excision induced by mastoparan without inhibiting rapid Ins(1,4,5)P3 production. Likewise, the Ins(1,4,5)P3-gated channel inhibitors ruthenium red and heparin blocked deflagellation in response to mastoparan. These studies were extended to mutants defective in flagellar excision. Fa-1, a mutant defective in flagellar structure, produced Ins(1,4,5)P3 but failed to deflagellate. These results support a model in which acid pH activates a putative cellular receptor leading to G-protein dependent activation of phospholipase C and accumulation of Ins(1,4,5)P3. These events are upstream of Ins(1,4,5)P3-dependent Ca2+ entry from the medium, and of deflagellation. PMID- 8227147 TI - Transgenic mouse models that explore the multistep hypothesis of intestinal neoplasia. AB - SV-40 T antigen (TAg), human K-rasVal12, and a dominant negative mutant of human p53 (p53Ala143) have been expressed singly and in all possible combinations in postmitotic enterocytes distributed throughout the duodenal-colonic axis of 1-12 mo-old FVB/N transgenic mice to assess the susceptibility of this lineage to gene products implicated in the pathogenesis of human gut neoplasia. SV-40 TAg produces re-entry into the cell cycle. Transgenic pedigrees that produce K rasVal12 alone, p53Ala143 alone, or K-rasVal12 and p53Ala143 have no detectable phenotypic abnormalities. However, K-rasVal12 cooperates with SV-40 TAg to generate marked proliferative and dysplastic changes in the intestinal epithelium. These abnormalities do not progress to form adenomas or adenocarcinomas over a 9-12-mo period despite sustained expression of the transgenes. Addition of p53Ala143 to enterocytes that synthesize SV-40 TAg and K rasVal12 does not produce any further changes in proliferation or differentiation. Mice that carry one, two, or three of these transgenes were crossed to animals that carry Min, a fully penetrant, dominant mutation of the Apc gene associated with the development of multiple small intestinal and colonic adenomas. A modest (2-5-fold) increase in tumor number was noted in animals which express SV-40 TAg alone, SV-40 TAg and K-rasVal12, or SV-40 TAg, K-rasVal12 and p53Ala143. However, the histopathologic features of the adenomas were not altered and the gut epithelium located between tumors appeared similar to the epithelium of their single transgenic, bi-transgenic, or tri-transgenic parents without Min. These results suggest that (a) the failure of the dysplastic gut epithelium of SV 40 TAg X K-rasVal12 mice to undergo further progression to adenomas or adenocarcinomas is due to the remarkable protective effect of a continuously and rapidly renewing epithelium, (b) initiation of tumorigenesis in Min mice typically occurs in crypts rather than in villus-associated epithelial cell populations, and (c) transgenic mouse models of neoplasia involving members of the enterocytic lineage may require that gene products implicated in tumorigenesis be directed to crypt stem cells or their immediate descendants. Nonetheless, directing K-rasVal12 production to proliferating and nonproliferating cells in the lower and upper half of small intestinal and colonic crypts does not result in any detectable abnormalities. PMID- 8227148 TI - Salmonella typhimurium attachment to human intestinal epithelial monolayers: transcellular signalling to subepithelial neutrophils. AB - In human intestinal disease induced by Salmonella typhimurium, transepithelial migration of neutrophils (PMN) rapidly follows attachment of the bacteria to the epithelial apical membrane. In this report, we model those interactions in vitro, using polarized monolayers of the human intestinal epithelial cell, T84, isolated human PMN, and S. typhimurium. We show that Salmonella attachment to T84 cell apical membranes did not alter monolayer integrity as assessed by transepithelial resistance and measurements of ion transport. However, when human neutrophils were subsequently placed on the basolateral surface of monolayers apically colonized by Salmonella, physiologically directed transepithelial PMN migration ensued. In contrast, attachment of a non-pathogenic Escherichia coli strain to the apical membrane of epithelial cells at comparable densities failed to stimulate a directed PMN transepithelial migration. Use of the n-formyl-peptide receptor antagonist N-t-BOC-1-methionyl-1-leucyl-1- phenylalanine (tBOC-MLP) indicated that the Salmonella-induced PMN transepithelial migration response was not attributable to the classical pathway by which bacteria induce directed migration of PMN. Moreover, the PMN transmigration response required Salmonella adhesion to the epithelial apical membrane and subsequent reciprocal protein synthesis in both bacteria and epithelial cells. Among the events stimulated by this interaction was the epithelial synthesis and polarized release of the potent PMN chemotactic peptide interleukin-8 (IL-8). However, IL-8 neutralization, transfer, and induction experiments indicated that this cytokine was not responsible for the elicited PMN transmigration. These data indicate that a novel transcellular pathway exists in which subepithelial PMN respond to lumenal pathogens across a functionally intact epithelium. Based on the known unique characteristics of the intestinal mucosa, we speculate that IL-8 may act in concert with an as yet unidentified transcellular chemotactic factor(s) (TCF) which directs PMN migration across the intestinal epithelium. PMID- 8227149 TI - Human osteogenic protein-1 induces both chondroblastic and osteoblastic differentiation of osteoprogenitor cells derived from newborn rat calvaria. AB - Osteogenetic protein-1 (OP-1), a member of the TGF-beta superfamily, induces endochondrial bone formation at subcutaneous sites in vivo and stimulates osteoblastic phenotypic expression in vitro. Primary cultures of newborn rat calvarial cells contain a spectrum of osteogenic phenotypes ranging from undifferentiated mesenchymal osteoprogenitor cells to parathyroid hormone (PTH) responsive osteoblasts. We examined whether treatment of this cell population with recombinant human osteogenic protein-1 could induce chondrogenesis in vitro. Markers of chondroblastic versus osteoblastic differentiation included alcian blue staining at pH 1, alkaline phosphatase-specific activity, osteocalcin radioimmunoassay, and expression of collagen mRNAs. 6 d of treatment (culture days 1-7) with 4-100 ng OP-1/ml caused dose-dependent increases in alcian blue staining intensity and alkaline phosphatase activity (4.7- and 3.4-fold, respectively, at 40 ng/ml), while osteocalcin production decreased twofold. Clusters of round, refractile, alcian blue-stained cells appeared by day 3, increased in number until day 7, and then became hypertrophic and gradually became less distinct. Histochemically, the day 7 clusters were associated with high alkaline phosphatase activity and became mineralized. mRNA transcripts for collagen types II and IX were increased by OP-1, peaking at day 4, while type X collagen mRNA was detectable only on day 7 in OP-1-treated cultures. Delay of OP 1 exposure until confluence (day 7) amplifies expression of the normal osteoblastic phenotype and accelerates its developmental maturation. In contrast, early OP-1 treatment commencing on day 1 strongly amplifies chondroblastic differentiation. In the same protocol, TGF-beta 1 alone at 0.01-40 ng/ml fails to induce any hypertrophic chondrocytes, and in combination with OP-1, TGF-beta 1 blocks OP-1-dependent chondroinduction. OP-1 is believed to act on a subpopulation of primitive osteoprogenitor cells to induce endochondrial ossification, but does not appear to reverse committed osteoblasts to the chondrocyte phenotype. PMID- 8227150 TI - Accumulation of actin in subsets of pioneer growth cone filopodia in response to neural and epithelial guidance cues in situ. AB - Directed outgrowth of neural processes must involve transmission of signals from the tips of filopodia to the central region of the growth cone. Here, we report on the distribution and dynamics of one possible element in this process, actin, in live growth cones which are reorienting in response to in situ guidance cues. In grasshopper embryonic limbs, pioneer growth cones respond to at least three types of guidance cues: a limb axis cue, intermediate target cells, and a circumferential band of epithelial cells. With time-lapse imaging of intracellularly injected rhodamine-phalloidin and rhodamine-actin, we monitored the distribution of actin during growth cone responses to these cues. In distal limb regions, accumulation of actin in filopodia and growth cone branches accompanies continued growth, while reduction of actin accompanies withdrawal. Where growth cones are reorienting to intermediate target cells, or along the circumferential epithelial band, actin selectively accumulates in the proximal regions of those filopodia that have contacted target cells or are extending along the band. Actin accumulations can be retrogradely transported along filopodia, and can extend into the central region of the growth cone. These results suggest that regulation and translocation of actin may be a significant element in growth cone steering. PMID- 8227151 TI - Identification and differential subcellular localization of the neuronal class C and class D L-type calcium channel alpha 1 subunits. AB - To identify and localize the protein products of genes encoding distinct L-type calcium channels in central neurons, anti-peptide antibodies specific for the class C and class D alpha 1 subunits were produced. Anti-CNC1 directed against class C immunoprecipitated 75% of the L-type channels solubilized from rat cerebral cortex and hippocampus. Anti-CND1 directed against class D immunoprecipitated only 20% of the L-type calcium channels. Immunoblotting revealed two size forms of the class C L-type alpha 1 subunit, LC1 and LC2, and two size forms of the class D L-type alpha 1 subunit, LD1 and LD2. The larger isoforms had apparent molecular masses of approximately 200-210 kD while the smaller isoforms were 180-190 kD, as estimated from electrophoresis in gels polymerized from 5% acrylamide. Immunocytochemical studies using CNC1 and CND1 antibodies revealed that the alpha 1 subunits of both L-type calcium channel subtypes are localized mainly in neuronal cell bodies and proximal dendrites. Relatively dense labeling was observed at the base of major dendrites in many neurons. Staining in more distal dendritic regions was faint or undetectable with CND1, while a more significant level of staining of distal dendrites was observed with CNC1, particularly in the dentate gyrus and the CA2 and CA3 areas of the hippocampus. Class C calcium channels were concentrated in clusters, while class D calcium channels were generally distributed in the cell surface membrane of cell bodies and proximal dendrites. Our results demonstrate multiple size forms and differential localization of two subtypes of L-type calcium channels in the cell bodies and proximal dendrites of central neurons. The differential localization and multiple size forms may allow these two channel subtypes to participate in distinct aspects of electrical signal integration and intracellular calcium signaling in neuronal cell bodies. The preferential localization of these calcium channels in cell bodies and proximal dendrites implies their involvement in regulation of calcium-dependent functions occurring in those cellular compartments such as protein phosphorylation, enzyme activity, and gene expression. PMID- 8227152 TI - R2D5 antigen: a calcium-binding phosphoprotein predominantly expressed in olfactory receptor neurons. AB - R2D5 is a mouse monoclonal antibody that labels rabbit olfactory receptor neurons. Immunoblot analysis showed that mAb R2D5 recognizes a 22-kD protein with apparent pI of 4.8, which is abundantly contained in the olfactory epithelium and the olfactory bulb. We isolated cDNA for R2D5 antigen and confirmed by Northern analysis and neuronal depletion technique that R2D5 antigen is expressed predominantly, but not exclusively, in olfactory receptor neurons. Analysis of the deduced primary structure revealed that R2D5 antigen consists of 189 amino acids with calculated M(r) of 20,864 and pI of 4.74, has three calcium-binding EF hands, and has possible phosphorylation sites for Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) and cAMP-dependent protein kinase (A kinase). Using the bacterially expressed protein, we directly examined the biochemical properties of R2D5 antigen. R2D5 antigen binds Ca2+ and undergoes a conformational change in a manner similar to calmodulin. R2D5 antigen is phosphorylated in vitro by CaM kinase II and A kinase at different sites, and 1.81 and 0.80 mol of Pi were maximally incorporated per mol of R2D5 antigen by CaM kinase II and A kinase, respectively. Detailed immunohistochemical study showed that R2D5 antigen is also expressed in a variety of ependymal cells in the rabbit central nervous system. Aside from ubiquitous calmodulin, R2D5 antigen is the first identified calcium-binding protein in olfactory receptor neurons that may modulate olfactory signal transduction. Furthermore our results indicate that olfactory receptor neurons and ependymal cells have certain signal transduction components in common, suggesting a novel physiological process in ependymal cells. PMID- 8227153 TI - Integrin-cytoskeletal interactions in migrating fibroblasts are dynamic, asymmetric, and regulated. AB - We have used laser optical trapping and nanometer-level motion analysis to investigate the cytoskeletal associations and surface dynamics of beta 1 integrin, a cell-substrate adhesion molecule, on the dorsal surfaces of migrating fibroblast cells. A single-beam optical gradient trap (laser tweezers) was used to restrain polystyrene beads conjugated with anti-beta 1 integrin mAbs and place them at desired locations on the cell exterior. This technique was used to demonstrate a spatial difference in integrin-cytoskeleton interactions in migrating cells. We found a distinct increase in the stable attachment of beads, and subsequent rearward flow, on the lamellipodia of locomoting cells compared with the retracting portions. Complementary to the enhanced linkage of integrin at the cell lamellipodium, the membrane was more deformable at the rear versus the front of moving cells while nonmotile cells did not exhibit this asymmetry in membrane architecture. Video microscopy and nanometer-precision tracking routines were used to study the surface dynamics of integrin on the lamellipodia of migrating cells by monitoring the displacements of colloidal gold particles coated with anti-beta 1 integrin mAbs. Small gold aggregates were rapidly transported preferentially to the leading edge of the lamellipod where they resumed diffusion restricted along the edge. This fast transport was characterized by brief periods of directed movement ("jumps") having an instantaneous velocity of 37 +/- 15 microns/min (SD), separated by periods of diffusion. In contrast, larger aggregates of gold particles and the large latex beads underwent slow, steady rearward movement (0.85 +/- 0.44 micron/min) (SD) at a rate similar to that reported for other capping events and for migration of these cells. Cell lines containing mutated beta 1 integrins were used to show that the cytoplasmic domain is essential for an asymmetry in attachment of integrin to the underlying cytoskeletal network and is also necessary for rapid, intermittent transport. However, enhanced membrane deformability at the cell rear does not require integrin-cytoskeletal interactions. We also demonstrated that posttranslational modifications of integrin could potentially play a role in these phenomena. These results suggest a scheme for the role of dynamic integrin mediated adhesive interactions in cell migration. Integrins are transported preferentially to the cell front where they form nascent adhesions. These adhesive structures grow in size and associate with the cytoskeleton that exerts a rearward force on them. Dorsal aggregates more rearward while those on the ventral side remain fixed to the substrate allowing the cell body to move forward. Detachment of the cell rear occurs by at least two modes: (a) weakened integrin-cytoskeleton interactions, potentially mediated by local modifications of linkage proteins, which lead to weakened cell-substratum interactions and (b) ripping of integrins and the highly deformable membrane from the cell body. PMID- 8227155 TI - Involvement of the conserved acidic amino acid domain of FGF receptor 1 in ligand receptor interaction. AB - The fibroblast growth factor receptor 1 (flg) contains eight acidic amino acids between the first and second immunoglobulin domain. This report examines the role of the acidic domain in the interaction of the flg receptor with its ligands. We observed a marked inhibition of binding of bFGF to the receptor when the acidic domain was completely deleted, but mutants with two and four amino acids deleted (flg delta A2 and flg delta A4, respectively) still bound the ligand. After addition of a bifunctional cross-linking reagent, cross-linked complexes (between bFGF and receptor) with the expected size were observed in cells expressing mutants lacking two or four acidic residues, but not in cells expressing mutants lacking six or eight acidic residues. Immunoprecipitation with anti-flg antibody followed by electrophoresis produced a band of 90 Kd in tunicamycin-treated cells expressing the mutant as well as the wild-type receptors, indicating that the inhibition of binding was not due to defective expression of the protein. The ability of flg delta A8 to mediate a mitogenic response to FGFs was also greatly reduced when this mutated receptor was expressed in receptor-negative cells. The effect of replacing the acidic amino acids with lysine residues was also studied. Binding of bFGF to cells transfected with a plasmid encoding a mutated protein with four amino acid substitutions was totally inhibited, but an eight amino acid substitution did not alter ligand binding to the receptor. In this case the mutation with four amino acids substitution caused a drastic impairment of protein expression. Thus the acidic domain of the FGFR-1 plays an essential role in receptor function, either because it is important for a stable protein configuration or for ligand-receptor interaction. PMID- 8227154 TI - Identification of sequences required for the efficient localization of the focal adhesion kinase, pp125FAK, to cellular focal adhesions. AB - The integrin family of heterodimeric cell surface receptors play critical roles in multiple biological processes by mediating cellular adhesion to the extracellular matrix (ECM). Adhesion triggers intracellular signaling cascades, including tyrosine phosphorylation and elevation of [Ca2+]i. The Focal Adhesion Kinase (FAK or pp125FAK), a protein tyrosine kinase that colocalizes with integrins in cellular focal adhesions, is a prime candidate for a mediator of integrin signaling events. Here we report an analysis of the domain structure of FAK in which we have identified a contiguous stretch of 159 amino acids within the COOH terminus essential for correct subcellular localization. When placed in the context of an unrelated cytosolic protein, this Focal Adhesion Targeting (FAT) sequence functions to efficiently mediate the focal adhesion localization of this fusion protein. Furthermore, this analysis suggests that pp125FAK cannot be activated oncogenically by mutation. This result could be explained if pp125FK either exhibits a narrow substrate specificity or is diametrically opposed by cellular phosphatases or other cellular processes. PMID- 8227156 TI - Insulin induces rapid accumulation of insulin receptors and increases tyrosine kinase activity in the nucleus of cultured adipocytes. AB - To better understand the mechanism by which insulin exerts effects on events at the cell nucleus, we have studied insulin receptors and tyrosine kinase activity in nuclei isolated by sucrose density gradient centrifugation following insulin treatment of differentiated 3T3-F442A cells. Insulin stimulated nuclear accumulation of insulin receptors by approximately threefold at 5 min. The half maximal effect was observed with 1-10 nM insulin. Following insulin treatment, phosphotyrosine content associated with the nuclear insulin receptor was also increased by twofold at 5 min with a similar insulin concentration dependency. These nuclear insulin receptors differ from the membrane-associated insulin receptors in that they were not efficiently solubilized with 1% Triton X-100. During the same period of time, insulin stimulated nuclear tyrosine kinase activity toward the exogenous substrate poly Glu4:Tyr1 tenfold in a time dependent manner reaching a maximum at 30 min. The insulin receptor substrate protein 1 (IRS-1) could not be detected in the nucleus by immunoblotting. However, a nuclear protein with M(r) approximately 220 kDa was tyrosine phosphorylated, and insulin further stimulated this process threefold > 30 mins. Surface labeling was performed to determine if the nuclear insulin receptors would emerge from the plasma membrane fraction. Using 125I-BPA-insulin with intact cells, the intensity of nuclear insulin receptor labeling was negligible and not increased throughout 30 min incubation at 37 degrees C. In contrast, there was an increase in labeled receptors in the microsomal fraction following insulin treatment. Taken together, these results indicate that insulin rapidly increases nuclear insulin receptor appearance and activates nuclear tyrosine kinase activity. The insulin-induced accumulation of nuclear insulin receptors cannot be accounted for by internalization of surface membrane receptors. These effects of insulin may play an important role in action of the hormone at the nuclear level. PMID- 8227157 TI - Cell substratum modulates responses of preosteoblasts to retinoic acid. AB - The aim of this study was to determine the role of ECM components of bone in regulating the differentiation and function of cells of the osteoblast lineage. Rat UMR 201 cells, phenotypically preosteoblast, were plated onto plastic tissue culture dishes or dishes coated with gelled type I collagen or reconstituted basement membrane (matrigel). Acute cell attachment assays showed that cells adhered to substrates in the following order: collagen > matrigel >> plastic. Proliferation rate up to 96 hr were similar on each substrate. However, if cells were treated with 10(-6) M retinoic acid (RA), proliferation rates were reduced compared with control for cells grown on collagen and matrigel but not on plastic. Morphological changes were matrix-specific; in subconfluent cultures, long thin processes were seen with cells grown on collagen and a pattern of interconnecting cell processes formed when cells were plated on matrigel. Striking differences were observed in the constitutive or RA-induced gene expression of cells grown on the different substrates. When cells plated on collagen were treated with RA, induction of mRNA for alkaline phosphatase (ALP) as well as ALP enzyme activity were much less than with cells grown on plastic. In contrast, RA treatment induced osteopontin (OP) mRNA expression more strongly in cells plated on collagen compared with plastic within 24 hr and this was maintained for 72 hr. RA treatment produced a two fold increase of pro-alpha 1(I) collagen mRNA in cells grown on plastic and matrigel but not in cells grown on collagen. Growth on collagen produced changes in the way UMR 201 cells responded to RA from which they did not fully recover in subsequent 48-hr growth periods on plastic. These results indicate that ECM components regulate the function of and are capable of modulating RA-induced differentiation of preosteoblasts. PMID- 8227158 TI - Down-modulation of epidermal growth factor receptor accompanies TNF-induced differentiation of the DiFi human adenocarcinoma cell line toward a goblet-like phenotype. AB - Although the biologic response modifier tumor necrosis factor-alpha (TNF) is a known differentiation inducer in hematopoietic cells, its role in differentiation of other tissue types has yet to be elucidated. In the studies presented here, TNF treatment of the human rectal adenocarcinoma cell line, DiFi, elicits characteristics of early stage differentiating, mucin-producing colonocytes. Not only are TNF-treated DiFi cells growth-inhibited by TNF, but they also display a unique morphology. Additionally, TNF treatment of DiFi cells enhances > fivefold the expression of high molecular weight mucin glycoproteins, as measured by [125I]-wheat germ agglutinin (WGA) binding and the human milk fat globule-1 (HMFG 1) anti-MUC1 antibody reactivity. The induction of these differentiation characteristics correlates with novel alterations in epidermal growth factor receptor (EGF-R). Following 5-day TNF treatment of DiFi cultures, EGF receptor levels, kinase autophosphorylation activity, and receptor tyrosine phosphorylation are reduced by > fourfold. The establishment of a model system in which goblet-like cell characteristics and alterations in a growth factor receptor can be induced in vitro may be potentially useful in studying the underlying mechanisms of colonic epithelial cell proliferation and differentiation. PMID- 8227159 TI - Different patterns of apoptosis of HL-60 cells induced by cycloheximide and camptothecin. AB - Cells of the human promyelocytic HL-60 line, when treated with a variety of antitumor agents in the presence of the protein synthesis inhibitor cycloheximide (CHX), or with CHX alone, rapidly undergo apoptosis ("active cell death"). It is presumed, therefore, that such cells are "primed" to apoptosis in that no new protein synthesis is required for induction of their death. We have studied apoptosis of HL-60 cells triggered by the DNA topoisomerase I inhibitor camptothecin (CAM) in the absence and presence of CHX and apoptosis induced by CHX alone. Two different flow cytometric methods were used, each allowing us to relate the apoptosis-associated DNA degradation to the cell cycle position. Apoptosis induced by CAM was limited to S phase cells, e.g., at a CAM concentration of 0.15 microM, nearly 90% of the S phase cells underwent apoptosis after 4 h. In contrast, apoptosis triggered by CHX was indiscriminate, affecting all phases of the cycle: approximately 40% of the cells from each phase the cycle underwent apoptosis at 5 microM CHX concentration. When CAM and CHX were added together, the pattern of apoptosis resembled that of cycloheximide alone, namely, cells in all phases of the cycle in similar proportion were affected. Thus, CHX, while itself inducing apoptosis of a fraction of cells, protected the S phase cells against apoptosis triggered by CAM. Because CHX (5 microM) did not significantly affect the rate of cell progression through S phase, the observed protective effect was most likely directly related to inhibition of protein synthesis, rather than to its possible indirect effect on DNA replication. Furthermore, whereas apoptosis (DNA degradation) triggered by CAM was prevented by the serine protease inhibitor N-tosyl-L-lysylchloromethyl ketone (TLCK), this process was actually potentiated by this inhibitor when induced by CHX. The present data indicate differences in mechanism of apoptosis triggered by CAM (and perhaps other antitumor drugs) as compared with CHX. Apoptosis caused by CHX may be unique in that it may not involve new protein synthesis. These data are compatible with the assumption that the loss of a hypothetical, rapidly turning over suppressor of apoptosis may be the trigger of apoptosis of HL-60 cells treated with CHX, whereas de novo protein synthesis is required when apoptosis is triggered by other agents. PMID- 8227160 TI - Maturation-dependent regulation of protein kinase C activity by vitamin D3 metabolites in chondrocyte cultures. AB - Vitamin D3 metabolites regulate the differentiation of chondrocytes isolated from the growth zone or resting zone of rat costochondral cartilage. Since some of the direct membrane effects of vitamin D metabolites are nongenomic, we hypothesized that protein kinase C (PKC) plays a role in signal transduction for these chondrocyte differentiation factors and that the regulation of PKC by the vitamin D metabolites is cell maturation dependent. Confluent, fourth passage cultures of growth zone and resting zone chondrocytes were treated with vitamin D3 metabolites for up to 24 h, lysed, and cell extracts assayed for kinase activity using a specific PKC substrate peptide. The addition of 1,25-(OH)2D3 to growth zone cell cultures resulted in a rapid dose-dependent stimulation of PKC, significant at 10(-9)-10(-7) M, beginning at 3 min and sustained until 90 min; 1,25-(OH)2D3 had no effect on PKC activity in resting zone chondrocyte cultures. The addition of 24,25-(OH)2D3 to resting zone cultures showed a slower PKC activation, with significant stimulation seen at 90-360 min for 10(-8)-10(-7) M 24,25-(OH)2D3. However, 24,25-(OH)2D3 had no effect on PKC activity in growth zone cell cultures at all times and concentrations examined. The specificity of PKC stimulation by the vitamin D3 metabolites was verified using a specific pseudosubstrate region peptide inhibitor, which reduced PKC activity when included in the reaction mixture. Pretreatment of the cultures with U73, 122, a phospholipase C inhibitor, decreased 1,25-(OH)2D3-stimulated PKC activity but had no effect upon 24,25-(OH)2D3-induced activity. The tyrosine kinase inhibitor, genistein, did not inhibit the PKC response in either vitamin D3 metabolites treated culture. Neither actinomycin D nor cycloheximide affected 1,25-(OH)2D3 induced PKC activity in growth zone chondrocyte cultures, while both compounds inhibited 24,25-(OH)2D3-induced activity in resting zone chondrocyte cultures. The results of this study indicate that vitamin D metabolites stimulate PKC activity in a metabolite- and cell-maturation-specific manner. Effects of 1,25 (OH)2D3 appear to be nongenomic, whereas the effects of 24,25-(OH)2D3 probably involve a genomic mechanism. PMID- 8227161 TI - Senescence of aortic endothelial cells in culture: effects of basic fibroblast growth factor expression on cell phenotype, migration, and proliferation. AB - Bovine aortic endothelial cells (BAEC) can be isolated in large numbers without major contamination by other cells and maintained in culture with a limited life span for about 100 population doublings. In order to study phenotypic changes of BAEC during long-term culture, stocks of different passages of BAEC were established and their morphological, migratory, and proliferative properties analyzed. Early-passage BAEC (passages 5-15) rapidly produce dense, cobblestone like monolayers. Their growth beyond the monolayer configuration is characterized by the formation of an irregular network of spindle-shaped, crisscrossing BAEC growing either on top or beneath the monolayer, and by the assembly of elongated BAEC into well-differentiated capillary-like tubes. In contrast, senescent BAEC (passages 35-45) form perfect cobblestone monolayers that contain several, often multinucleated giant cells and a few capillary-like tubes but not the crisscrossing networks of their early-passage counterparts. The rates of BAEC migration and proliferation gradually decline during in vitro senescence. This decline is neutralized by exogenous basic fibroblast growth factor (bFGF) which elevates the migratory and proliferative capacities of early-passage and senescent BAEC to uniformly high levels. Northern blot analysis shows a gradual decline in bFGF message and an increase in laminin message during in vitro BAEC senescence. The present study supports the concept of autocrine growth regulation of BAEC and associates a decreased bFGF message with decreased rates of migration and proliferation as well as loss of the crisscrossing BAEC morphotype in senescent cultures. PMID- 8227162 TI - Cell cycle activity and expression of prohibitin mRNA. AB - Prohibitin, a novel intracellular antiproliferative protein, blocks entry into the S phase of the cell division cycle when its mRNA is microinjected into normal fibroblasts or HeLa cells. To learn more about the interaction between prohibitin and the cell cycle, we studied the effect of microinjecting prohibitin mRNA at different points during the transition from G0 to S phase and analyzed prohibitin mRNA and protein levels in different parts of the cell cycle. The antiproliferative activity of microinjected prohibitin mRNA is high in G0/G1 and falls as cells approach S phase. Prohibitin mRNA and protein levels are high in G1, fall with S phase, rise again in G2, and fall in M. Together, these findings suggest that endogenous prohibitin contributes to the control of the G1 to S transition in cycling cells in a complex manner, which involves both a transcriptional and posttranslational mechanism. PMID- 8227163 TI - Transforming growth factor-beta, basic fibroblast growth factor, and platelet derived growth factor-BB interact to affect proliferation of clonally derived porcine satellite cells. AB - Transforming growth factor beta-1 (TGF-beta) stimulated porcine satellite cell proliferation in basal serum-free medium by 25%, but inhibited growth in serum containing medium by 58%. The effect of TGF-beta on cell proliferation in serum free medium was examined in combination with the following human recombinant growth factors: platelet-derived growth factor-BB (PDGF), basic fibroblast growth factor (FGF), insulin-like growth factor I (IGF-I), and epidermal growth factor (EGF). TGF-beta inhibited PDGF-stimulated proliferation, enhanced FGF-stimulated proliferation, and had no effect on proliferation stimulated by IGF-I. The response of satellite cells to EGF and TGF-beta in serum-free medium was not different than TGF-beta alone. TGF-beta depressed proliferation stimulated by the following combinations of two growth factors: PDGF and IGF-I, PDGF and EGF, PDGF and FGF, and IGF-I and EGF. In combination with IGF-I and FGF, TGF-beta did not affect proliferation. TGF-beta inhibited proliferation stimulated by the combination of PDGF, EGF, and IGF-I, but had no effect on proliferation stimulated by combinations of three growth factors that included FGF. FGF stimulated proliferation in Minimum Essential Medium containing 10% porcine serum (MEM-10% PS) by 13% above control. When the combination of TGF-beta and FGF was added to MEM-10% PS, a 78% increase in proliferation was observed. Polyclonal antihuman PDGF-AB (this form neutralizes PDGF-AA, AB, and BB) reduced proliferation in MEM-10% PS by 44%. The combination of TGF-beta and anti-PDGF-AB reduced proliferation by 59%, indicating the effects were not additive. These data indicate that: (1) FGF and TGF-beta interact to increase proliferation of clonally derived porcine satellite cells, and (2) the inhibitory effect of TGF beta on proliferation of clonally derived porcine satellite cells can be primarily attributed to a reduction in the mitogenic effects of PDGF. PMID- 8227164 TI - Fibroblast growth factor, epidermal growth factor, insulin-like growth factors, and platelet-derived growth factor-BB stimulate proliferation of clonally derived porcine myogenic satellite cells. AB - Clonally derived cultures of porcine skeletal muscle satellite cells were developed. The mitogenic effects of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulin-like growth factors (IGF-I and -II), and platelet-derived growth factors (PDGF-AA and -BB) were examined. Individually, bFGF, IGFs, and PDGF-BB stimulated proliferation of porcine satellite cells grown in basal serum-free medium or Minimum Essential Medium containing 2% fetal bovine serum (MEM-2% FBS). EGF stimulated proliferation in MEM-2% FBS, but neither EGF nor PDGF-AA were mitogenic when added to serum-free medium. The interactions among bFGF, EGF, IGF-I, and PDGF-BB were examined in serum-free medium, using growth factor concentrations shown in dose-response experiments to induce maximal proliferative responses (10 ng/ml bFGF, EGF and PDGF-BB, and 50 ng/ml IGF-I). The combination of bFGF and IGF-I dramatically increased proliferation, and IGF-I also synergized with EGF to increase proliferation. EGF, IGF-I, and bFGF interacted with PDGF-BB to stimulate proliferation. With the exception of EGF and bFGF, combinations of two growth factors typically resulted in greater than additive responses. Simultaneous exposure of satellite cells to bFGF, PDGF-BB, EGF, and IGF-I produced a fivefold increase in DNA compared to cells grown in basal serum-free medium. Elimination of EGF did not reduce the mitogenic response, yet removal of IGF-I, bFGF, or PDGF-BB reduced proliferation by approximately 40, 20, and 10%, respectively. These mitogens are likely physiological regulators of porcine satellite cell activity. PMID- 8227165 TI - Insulin receptor characterization and function in bovine aorta endothelial cells: insulin degradation by a plasma membrane, protease-resistant insulin receptor. AB - The functional significance of the insulin receptor on bovine aorta endothelial (BAE) cells is not well defined. The insulin receptor expressed on BAE cells does not mediate insulin hormonal effects and does not mediate the transcytosis of insulin from the apical to the basolateral domain of the cell monolayer. To assess the role of the insulin receptor on BAE cells, the physical characteristics of the BAE cell receptor were investigated, and the time dependent interaction of insulin and insulin degradation products with BAE cell monolayers was quantitated. The BAE cell insulin receptor was found to be highly resistant to the proteolytic action of trypsin, pronase, and proteinase K at either 4 degrees C or 37 degrees C. This resistance may permit the receptor to maintain insulin binding capabilities in spite of the high concentrations of proteases which are normally present in blood. Scatchard analysis of cell-surface and total cellular insulin receptor demonstrated dissociation constants similar to values obtained with other cells and tissues. However, whereas other cells and tissues contain an intracellular pool of receptor that ranges from 20-40% of the total cellular receptor content, no intracellular population of insulin receptors was detected in BAE cells. Upon incubation of intact BAE cell monolayers with insulin, no endocytosis of cell-surface insulin receptor could be demonstrated. However, insulin degradation by the BAE cells was readily quantitated, at a rate of 16.3 fmol/10(6) cells/h at an insulin concentration of 2 nM. This rate of degradation was not inhibited by chloroquine, which inhibits insulin degradation in fibroblasts, hepatocytes, and adipocytes, nor by phenylarsine oxide, which inhibits endocytosis. Bacitracin inhibited insulin binding to the cell monolayers and inhibited insulin degradation with identical IC50 values (80 microM). These data suggest that in BAE cells, insulin degradation occurs in the absence of receptor-mediated endocytosis and is mediated by binding of insulin to its receptor. Therefore, it is concluded that the functional role of the insulin receptor expressed in BAE cells is to bind blood-borne insulin at the plasma membrane of the cell and thereby facilitate the degradation of insulin at the BAE cell plasma membrane. PMID- 8227166 TI - MSH receptors in immortalized human epidermal keratinocytes: a potential mechanism for coordinate regulation of the epidermal-melanin unit. AB - Receptors for melanotropin (MSH) were found to be expressed by immortalized primary human epidermal keratinocytes (RHEK-1). Using 125I-beta MSH as a probe, the MSH receptors from mouse melanoma cells and human keratinocytes were found to be remarkably similar. In each cell line, there were high and low affinity receptors, with the high affinity classes showing positive cooperativity. Competition of 125I-beta MSH for binding with non-radioactive MSH revealed similar profiles. Cross-linking studies, followed by gel electrophoresis and autoradiography, showed almost identical gel migration patterns. Both cell types expressed internal as well as plasma membrane binding sites. MSH receptors on both cell types were up-regulated by ultraviolet light and by MSH itself. Although the function of MSH receptors expressed by the immortalized keratinocytes is unknown, the results are consistent with recent reports that proliferation of epidermal keratinocytes is stimulated by MSH and that proopiomelanocortin genes are expressed in the epidermis. These results support a model in which keratinocytes and melanocytes, interacting in an "epidermal melanin unit," each respond to UV light signals with increased MSH receptor activity. PMID- 8227167 TI - Specific simple sugars promote chemotaxis and chemokinesis of corneal endothelial cells. AB - Bovine corneal endothelial cells showed a strong migratory response to specific simple sugars (D-glucose and sucrose, but not L-glucose, sorbitol, lactose, or D galactose) at concentrations above 10 mM. Checkerboard analysis of the migratory responses in modified Boyden chambers indicated both chemotactic and chemokinetic effects. Serum starvation of the cultures increased the chemotaxis towards D glucose and 2-deoxy-D-glucose, but not towards sucrose. Migration to sucrose and glucose was inhibited by chelation of extracellular calcium or by inhibition of Na+, K+ ATPase with ouabain. To date, this migratory response has been found only in corneal endothelial cells. Neither human melanoma cells, human breast carcinoma cells, bovine aortic endothelial cells, nor bovine microvascular endothelial cells migrated towards simple sugars, although all cell types migrated toward fibronectin in chemotaxis assays. After 16-19 passages in culture, bovine corneal endothelial cells retained their ability to migrate towards fibronectin, but lost their ability to migrate towards sugars. This loss of migratory response was accompanied by a sevenfold decrease in Na+, K+ ATPase activity. Although loss of Na+, K+ ATPase activity accompanied the loss of migratory response, pretreatment of cell cultures with 25 mM glucose did not stimulate, but rather lowered Na+, K+ ATPase activity in low or high passage cultures. PMID- 8227168 TI - Differential regulation of actin polymerization following activation of resting T lymphocytes from young and aged mice. AB - Actin polymerization accompanies receptor-mediated responses and is correlated with motility-related events. In T lymphocytes, there is a lateral redistribution of surface receptors into caps and aggregation of actin-myosin in cytoplasmic subcaps, and these are impaired in T cells from aged individuals. This study documents marked changes in age-related cytoskeletal actin filament function which may account for the reduced motility. Basal levels of filamentous actin (F actin) are significantly higher in purified G(o) T cells from aged C57BL/6 mice, due to a preferential increase in the CD8+ subpopulation. Following activation of the resting T cells with Concanavalin A (Con A), F-actin depolymerized in cells from young mice for 2 min, followed by rapid polymerization, reaching a plateau 200% above resting levels. In cells from 15-17-month-old mice, an attenuated depolymerization phase was seen for 45 sec, followed by little polymerization. No depolymerization or polymerization phases occurred in cells from aged mice. Phorbol 12 myristate 13-acetate (PMA), which activates protein kinase C (PKC), bypassing receptor mediated signals, induced actin polymerization to 57% of the levels of that after Con A stimulation in cells from both young and old animals and partially eliminated the differences in actin filament assembly due to age. Perturbation of the cytoskeleton with cytochalasin E (CE) potentiated proliferation of Con A-stimulated T cells from aged mice but did not completely restore the deficit attributed to immunosenescence. The results show an age related impairment of cytoskeletal functions and suggest that differences in early signal transduction events contribute to the decrements in surface receptor motility and subsequent proliferation of T lymphocytes from older individuals. PMID- 8227169 TI - C-FMS dependent HL-60 cell differentiation and regulation of RB gene expression. AB - The dependence of induced myelomonocytic cell differentiation, and regulation of the RB tumor suppressor gene during this process, on the c-fms gene product, the CSF-1 lymphokine receptor, was determined in HL-60 promyelocytic leukemia cells. Adding a monoclonal antibody with specificity for the c-fms gene product to cells treated with various inducers of myelomonocytic or macrophage differentiation, including retinoic acid and 1,25-dihydroxy vitamin D3, inhibited the rate of differentiation. During the period of inducer treatment usually preceding onset of differentiation, longer periods of antibody exposure caused greater inhibition of differentiation. In a stable HL-60 transfectant overexpressing the CSF-1 receptor at the cell surface due to a constitutively driven c-fms trans gene, the rate of differentiation was enhanced compared to the wild type cell, consistent with a positive regulatory role for the CSF-1 receptor. The anti-fms antibody caused much less inhibition of differentiation in the transfectants than in wild type cells, consistent with a larger number of receptors causing reduced sensitivity. During the induced metabolic cascade leading to differentiation, the typical early down regulation of RB gene expression was inhibited by the antibody. The antibody itself caused an increase in RB expression per cell, which offset the decrease normally caused by differentiation inducers (1,25-dihydroxy vitamin D3 and retinoic acid). The changes in RB expression preceded changes in the RB protein to the hypophosphorylated state. Most of the RB protein in proliferating cells was phosphorylated and no significant accumulation of hypophosphorylated RB protein occurred until after onset of G0 arrest. Thus the metabolic cascade leading to myelomonocytic differentiation of HL-60 cells appears to be driven by a function of the c-fms protein. Inhibiting that process by attacking this receptor impedes differentiation and also compromises the early down regulation of RB tumor suppressor gene expression which normally precedes differentiation. These findings provide additional support for a potential role for down regulating RB expression in promoting cell differentiation, and suggest the possibility that RB may be either a target or intermediate mediator of CSF-1 actions. PMID- 8227170 TI - Low oxygen tension increases mRNA levels of alpha 1 (I) procollagen in human dermal fibroblasts. AB - Dermal fibroblasts exposed to low oxygen tension show upregulated synthesis of transforming growth factor-beta 1 (TGF-beta 1), an established stimulatory peptide in the formation of extracellular matrix proteins. In this report, procollagen synthesis was measured in cultures of confluent adult human dermal fibroblasts exposed to either standard (20%) or low (2%) oxygen tension. By Northern blot analysis the steady state levels of alpha 1 (I) procollagen mRNA were increased by 75 to 150% of control (standard oxygen) as early as 12 hours and more than 200% 96 hours after exposure of cells to low oxygen. Similar increases in procollagen mRNA levels were obtained in hypoxic fibroblast cultures in a collagen lattice. The stimulatory effect of hypoxia on procollagen mRNA levels in fibroblast monolayers was diminished by antibodies to TGF-beta, and could not be augmented further by the addition of TGF-beta 1, evidence that hypoxic fibroblasts may already be maximally stimulated by TGF-beta 1. We conclude that low oxygen tension enhances steady state mRNA levels of alpha 1 (I) procollagen, and that this effect is mediated at least in part by TGF-beta 1. PMID- 8227171 TI - Membrane-anchored proteoglycans of mouse macrophages: P388D1 cells express a syndecan-4-like heparan sulfate proteoglycan and a distinct chondroitin sulfate form. AB - Proteoglycan accumulation by thioglycollate-elicited mouse peritoneal macrophages and a panel of murine monocyte-macrophage cell lines has been examined to determine whether these cells express plasma membrane-anchored heparan sulfate proteoglycans. Initially, cells were screened for heparan sulfate and chondroitin sulfate glycosaminoglycans after metabolic labeling with radiosulfate. Chondroitin sulfate is secreted to a variable extent by every cell type examined. In contrast, heparan sulfate is all but absent from immature pre-monocytes and is associated predominantly with the cell layer of mature macrophage-like cells. In the P388D1 cell line, the cell-associated chondroitin sulfate is largely present as a plasma membrane-anchored proteoglycan containing a 55 kD core protein moiety, which appears to be unique. In contrast, the cell-associated heparan sulfate is composed of a proteoglycan fraction and protein-free glycosaminoglycan chains, which accumulate intracellularly. A fraction of the heparan sulfate proteoglycan contains a lipophilic domain and can be released from cells following mild treatment with trypsin, suggesting that it is anchored in the plasma membrane. Isolation of this proteoglycan indicates that it is likely syndecan-4: it is expressed as a heparan sulfate proteoglycan at the cell surface, it is cleaved from the plasma membrane by low concentrations of trypsin, and it consists of a single 37 kD core protein moiety that co-migrates with syndecan-4 isolated from NMuMG mouse mammary epithelial cells. Northern analysis reveals that a panel of macrophage-like cell lines accumulate similar amounts of syndecan-4 mRNA, demonstrating that this proteoglycan is expressed by a variety of mature macrophage-like cells. Syndecan-1 mRNA is present only in a subset of these cells, suggesting that the expression of this heparan sulfate proteoglycan may be more highly regulated by these cells. PMID- 8227172 TI - Induction of collagenase and stromelysin gene expression by mechanical injury in a vascular smooth muscle-derived cell line. AB - We describe a novel system for studying the production of matrix metalloproteinases types I and III (collagenase and stromelysin) by a vascular smooth muscle cell line (Rb-1 cells) in response to mechanical injury. Highly confluent Rb-1 cells are disrupted by passing a plastic comb around the plate to clear a series of circumferential paths, which are bordered by two ridges of displaced cells. Over the next 24 hours, cells migrate into the cleared areas. Northern blot analysis demonstrates the accumulation of mRNAs for collagenase and stromelysin during this process, although they are undetectable prior to injury. Cotreatment with all-trans-retinoic acid (10(-6) M) markedly decreases the level of mRNAs induced by injury, whereas dexamethasone (10(-7) M) causes only a slight reduction. In situ hybridization studies for stromelysin mRNA and immunohistochemical staining for collagenase protein on plates of injured cells showed the highest levels of stromelysin mRNA in cells in the ridges left by the injury; lower levels were observed in some cells migrating into the clear region. The same pattern of expression was observed when cells were stained with antiserum to collagenase protein. Nuclear run-on assays demonstrated increases in transcription of stromelysin and collagenase genes following injury. Transient transfection of cells with a vector containing the luciferase gene driven by a wild-type promoter comprising 1.8 kb of the 5'-flanking region of the rabbit collagenase gene showed increased activity associated with injury. We conclude that: (1) mechanical injury is associated with induction of mRNAs for the metalloproteinases collagenase and stromelysin, (2) retinoic acid effectively antagonizes this responses, and (3) the increase in steady-state mRNA levels is, at least in part, transcriptionally mediated. Thus our data suggest a role for mechanical forces in initiating the changes in gene expression in vascular smooth muscle cells following arterial injury in vivo. PMID- 8227173 TI - Association of a human H1 histone gene with an H2A pseudogene and genes encoding H2B.1 and H3.1 histones. AB - A cluster of human histone genes was found on three overlapping clones isolated from cosmid and bacteriophage libraries. These three overlapping segments of the human genome comprise genes coding for H3.1, an H2A pseudogene, and an H2B.1 gene downstream of the previously characterized H1.2 gene. The cosmid clone covers 30 kb upstream of the H1.2 gene and overlaps with two phage clones covering the core histone genes and the pseudogene. The same arrangement of an H3 gene, an H2A pseudogene and an H2B gene downstream of an H1 gene has been described within a mouse histone gene cluster [Yang et al.:J Biol Chem 262:17118-17125, 1987; Gruber et al.:Gene 95:303-304, 1990]. PMID- 8227174 TI - Gallium nitrate increases type I collagen and fibronectin mRNA and collagen protein levels in bone and fibroblast cells. AB - Gallium is a Group IIIa transitional element with therapeutic efficacy in the treatment of metabolic bone disorders. Previously described antiresorptive effects of gallium on osteoclasts are not sufficient to account for the full range of effects of gallium on bone structure and metabolism. We have recently shown that gallium nitrate inhibits osteocalcin gene expression and the synthesis of osteocalcin protein, an osteoblast-specific bone matrix protein that is thought to serve as a signal to trigger osteoclastic resorption. Here we present evidence for an additional mechanism by which gallium may function to augment bone mass by altering matrix protein synthesis by osteoblastic and fibroblastic cells. Rat calvarial explants exposed to gallium nitrate for 48 h showed increased incorporation of 3H-proline into hydroxyproline and collagenase digestible protein. In addition, gallium treatment increased steady-state mRNA levels for fibronectin and type I procollagen chains in primary rat calvarial osteoblast-enriched cultures, the ROS 17/2.8 osteoblastic osteosarcoma line, and nontransformed human dermal fibroblasts. These findings suggest that the exposure of mesenchymally-derived cells to gallium results in an altered pattern of matrix protein synthesis that would favor increased bone formation. PMID- 8227175 TI - Unambiguous identification of histone H1 in Trypanosoma cruzi. AB - The existence of histone H1 has been questioned in Trypanosomatids. We report here the presence of a histone H1 in the chromatin of Trypanosoma cruzi. This protein was purified by narrow-bore reversed phase HPLC and its amino acid composition analyzed and compared with histones H1 from other species. Furthermore, the purified chromosomal protein was digested with proteases and the amino acid sequences of the resulting peptides were analyzed by the automated Edman degradation. The sequences obtained were found to present a high degree of homology when compared to the carboxy terminal domain of other known histones H1. PMID- 8227176 TI - Phospholipase-induced maturation of Xenopus laevis oocytes: mitogenic activity of generated metabolites. AB - Signal transduction induced by generation of second messengers from membrane phospholipids is considered a major regulatory mechanism in control of cell proliferation. We report here that in the Xenopus laevis oocytes model, microinjection of the three most relevant types of phospholipases acting on membrane phospholipids (A2, C, and D) are capable of inducing oocyte maturation with similar efficiencies. This effect is mediated by the generation of known second messengers such as lyso-phospholipids, arachidonic acid, diacylglycerol, and phosphatidic acid. Specific inhibitors of protein kinase C made it possible to identify alternative independent signalling pathways for induction of oocyte maturation. Our results indicate that while phospholipase C seems to be dependent on protein kinase C (PKC), phospholipase A2, and phospholipase D are completely independent of protein kinase C function. Thus, the oocyte system is a powerful tool for the analysis of the potential mitogenic activity of lipid metabolites. It is also an excellent tool for unravelling the different routes involved in the regulation of cell growth. PMID- 8227177 TI - Identification and localization of G-proteins in the clonal adipocyte cell lines HGFu and Ob17. AB - HGFu and Ob17 are cell lines derived from adipose tissue of lean (+/?) and ob/ob mice, respectively. Neither adenylyl cyclase activity nor G protein abundance and subcellular distribution have been assessed previously in these cells. Cyclase activity was low and resistant to catecholamine stimulation in both cell lines. However, the enzyme could be stimulated to high levels by forskolin and Mn2+. Gs alpha (largely the long isoform), Gi alpha 2, and G beta were the major G protein subunits identified. The levels of G protein mRNA expression were similar in both cell lines and, unlike actin expression, did not change as a result of differentiation. Immunoblotting and ADP-ribosylation of the G peptides corroborated these results. Assessment of the subcellular localization of the subunits by indirect epifluorescence and scanning confocal microscopy showed that each of the subunits had a characteristic subcellular pattern. Gs alpha showed vesicular cytoplasmic and nuclear staining; Gi alpha 2 colocalized with actin stress fibers and disruption of these structures altered the distribution of Gi alpha 2; beta subunits showed some colocalization with the stress fibers as well as a cytoplasmic vesicular and nuclear pattern. As a result of differentiation, there was reorganization of the actin, together with the Gi alpha 2 and beta fibrous patterns. Both cell lines showed similar modifications. The induction of differentiation in these cells is therefore not associated with changes in adenylyl cyclase activity nor of the abundance of G-protein subunits, although reorganization of some of these subunits does accompany actin reorganization. PMID- 8227178 TI - Multitude of inverted repeats characterizes a class of anchorage sites of chromatin loops to the nuclear matrix. AB - In order to understand the nature of DNA sequences that organize chromatin into domains or loops, we have cloned the nuclear matrix DNA (1.7% of the total DNA) from human myelogenous leukemia cells in culture. Nuclear matrix is formed by interactions between specific stretches of DNA of about 0.1 to 5.0 kb with protein transcription factors, nuclear enzymes, and structural proteins. Nuclear matrix is believed to be the exclusive nuclear microenvironment in which initiation of DNA replication, transcription, and repair take place. The matrix attachment regions (MARs) of DNA have transcriptional enhancer activity, harbor the origins of replication of the human genome, and define the borders between neighboring chromatin loops. In this study we report the sequence of the human MAR fragment 19.2 of a size of 542 bp. Hum. MAR 19.2 is composed of TG-, CA-, CT , and GA-rich blocks and shows 8 perfect and imperfect inverted repeats. Thus, we have identified a novel class of MARs with sequence characteristics divergent from the AT-rich class of MARs. The inverted repeats of the 19.2 sequence might be stabilized into their cruciform configuration by torsional strain and by specific transcription/replication protein factors. This MAR might function in the initiation of replication of the flanking chromatin domain and in the regulation of the transcriptional activity of the gene(s) that reside in this domain. PMID- 8227179 TI - Arrest of cell cycle progression of HeLa cells in the early G1 phase in K(+) depleted conditions and its recovery upon addition of insulin and LDL. AB - Cell cycle progression of synchronized HeLa cells was studied by measuring labeling of the nuclei with [3H]thymidine. The progression was arrested in a chemically defined medium in which K+ was replaced by Rb+ (Rb-CDM) but was restored upon addition of insulin and/or low density lipoprotein (LDL). Cells started DNA synthesis 12 hr after addition of insulin and/or LDL, regardless of the time of arrest, suggesting their arrest early in the G1 phase. After incubation of cells in Rb-CDM containing insulin or LDL singly for 3, 6, or 9 hr, replacement of the medium by that without an addition resulted in marked delay in entry of cells into the S phase, but in its replacement by medium containing both agents, the delay was insignificant. Synthesis of bulk protein, estimated as increase in the cell volume, was not strongly inhibited. From these results we conclude that cell cycle progression of HeLa cells in K(+)-depleted CDM is arrested early in the G1 phase and that the arrest is due to lack of some protein(s) required for entry into the S phase that is synthesized in the early G1 phase. PMID- 8227180 TI - Density-dependent endothelial cell production of an inhibitor of smooth muscle cell growth. AB - Embryonic data and ultrastructural analyses suggest that the primitive endothelium signals undifferentiated mesenchymal cells to migrate to the forming blood vessel and subsequently regulates mural cell growth and behavior. Upon maturation of the blood vessel, chemotactic and mitogenic signals are apparently diminished and differentiated smooth muscle cells normally remain quiescent. This homeostasis is seemingly upset in conditions which lead to pathologies characterized by smooth muscle cell hyperplasia such as atherosclerosis. By culturing endothelial cells at different densities, we attempted to re-create the various stages of vascular development. Whereas media conditioned by sparse endothelial cells stimulate smooth muscle cells, media conditioned by dense endothelial cell cultures are inhibitory. Culture of sparse smooth muscle cells in media conditioned for 3 days by postconfluent endothelial cell cultures leads to dose-dependent and reversible smooth muscle cell inhibition. Furthermore, in the presence of the endothelial cell-derived inhibitor, smooth muscle cells are rendered refractory to mitogens such as fibroblast growth factor and platelet derived growth factor. The inhibitory activity is not attributable to the well characterized inhibitors of smooth muscle cell growth, transforming growth factor type-beta, prostaglandin I2, or heparan sulfate proteoglycan. Partial characterization of the inhibitory conditioned media suggests that the active molecule is smaller than 1,000 da, and stable to boiling as well as proteinase K and heparinase digestion. These findings support the concept that there is intercellular communication between endothelial cells and smooth muscle cells and provide evidence for a novel endothelial cell-derived smooth muscle cell growth inhibitor. PMID- 8227181 TI - Increase in epidermal growth factor receptor and its mRNA levels by parathyroid hormone (1-34) and parathyroid hormone-related protein (1-34) during differentiation of human trophoblast cells in culture. AB - Human cytotrophoblasts in culture aggregate and fuse to form syncytiotrophoblasts. This process is associated with an increase in epidermal growth factor receptor (EGFR) expression [Alsat et al.: J Cell Physiol 154:122 128, 1993]. Recent studies have demonstrated the presence of parathyroid hormone related protein (PTHrP) in the human uterus and placenta. This led us to study the effect of PTH (1-34) and PTHrP (1-34) on the expression of EGFR during this differentiation process. Both peptides induced a concentration-dependent increase in EGF binding, with a maximal effect at the physiological concentration of 1 nM. EGFR protein level assessed by cross-linking and immunoblotting and EGFR biological activity assessed by measuring its EGF-induced autophosphorylation were increased 2- and 2.5-fold, respectively, when cells were treated for 24 h with 0.1 microM PTHrP or PTH compared to control cells. This effect was time dependent with a maximum at 3 h of treatment. This treatment also increased trophoblast cell EGFR mRNA levels, suggesting transcriptional regulation of the EGFR. To ascertain whether activation of protein kinase C (PKC) or protein kinase A (PKA) is involved in this PTH effect, we determined EGFR protein level and EGFR autophosphorylation after exposure of cells to PKA inhibitor and PKC inhibitor, alone or together with the peptide. The presence of a PKC inhibitor blocked a further increase in EGFR number by PTH, while PKA inhibitor had no effect. These results show that PTH and PTHrP increase the synthesis of EGF receptors which are strongly expressed in syncytiotrophoblasts and suggested that these peptides might be involved in human placental development. PMID- 8227182 TI - Evidence against a role for alkaline phosphatase in the dephosphorylation of plasma membrane proteins: hypophosphatasia fibroblast study. AB - A major impasse to understanding the physiologic role(s) of alkaline phosphatase (ALP) is uncertainty as to its natural substrates. Various in vitro studies have led other investigators to suggest that ALP functions as a plasma membrane phosphoprotein phosphatase, consistent with our demonstration of ecto-topography of ALP in a variety of cell types. Thus, we compared the phosphorylation of plasma membrane proteins from control fibroblasts to those from profoundly ALP deficient fibroblasts of hypophosphatasia patients. Fibroblasts from 3 controls and 3 hypophosphatasia patients (ALP activity < 4% of control) were biosynthetically labeled with 32Pi for 2 h. 32P incorporation into total trichloroacetic acid (TCA)-precipitable material was not significantly different in control and patient cells. Plasma membranes were prepared from these cells by hypotonic shock, solubilized, and subjected to two-dimensional (2-D) gel electrophoretic separation. Video densitometric analysis of silver-stained 2-D gels failed to reveal any consistent difference in the protein profile between patient vs. control fibroblasts (i.e., unique species, altered pls, or increased abundance). Autoradiography of individual 2-D gels demonstrated 63 plasma membrane phosphoproteins with molecular weights ranging from 15 to 152 kDa and predominantly acidic pls. Although several of these phosphoproteins appeared to have had donor-specific labeling, none was unique or especially abundant in the hypophosphatasia group. Thus, in ALP-deficient fibroblasts, normal incorporation of 32P into total cellular protein and into all identifiable plasma membrane phosphoproteins indicates that ALP does not modulate the phosphorylation of plasma membrane proteins. PMID- 8227183 TI - Modulation of endothelial cell proliferation, adhesion, and motility by recombinant heparin-binding domain and synthetic peptides from the type I repeats of thrombospondin. AB - Thrombospondin is an inhibitor of angiogenesis that modulates endothelial cell adhesion, proliferation, and motility. Synthetic peptides from the second type I repeat of human thrombospondin containing the consensus sequence-Trp-Ser-Pro-Trp- and a recombinant heparin binding fragment from the amino-terminus of thrombospondin mimic several of the activities of the intact protein. The peptides and heparin-binding domain promote endothelial cell adhesion, inhibit endothelial cell chemotaxis to basic fibroblast growth factor (bFGF), and inhibit mitogenesis and proliferation of aortic and corneal endothelial cells. The peptides also inhibit heparin-dependent binding of bFGF to corneal endothelial cells. The antiproliferative activities of the peptides correlate with their ability to bind to heparin and to inhibit bFGF binding to heparin. Peptides containing amino acid substitutions that eliminate heparin-binding do not alter chemotaxis or proliferation of endothelial cells. Inhibition of proliferation by the peptide is time-dependent and reversible. Thus, the antiproliferative activities of the thrombospondin peptide and recombinant heparin-binding domain result at least in part from competition with heparin-dependent growth factors for binding to endothelial cell proteoglycans. These results suggest that both the Trp-Ser-Xaa-Trp sequences in the type I repeats and the amino-terminal domain play roles in the antiproliferative activity of thrombospondin. PMID- 8227184 TI - Studies of the levamisole inhibitory effect on rat stromal-cell commitment to mineralization. AB - The ability of Levamisole to decrease mineralization in skeletal tissue is usually related to its effect on alkaline phosphatase (ALP). However, Levamisole is also suspected to diminish mineralization by an additional mechanism which is unrelated to the ALP control of apatite crystal growth. To delineate the time in differentiation during which Levamisole inhibits mineralization, a tissue culture model system of bone marrow stromal cells was used. Secondary cultures of stromal cells were propagated in osteoprogenitor cell (OPC) induction medium for three weeks, followed by measurement of calcium precipitation. In situ ALP assays at pH 7.6 were also performed. When cells were cultured with 0.2 mM Levamisole for three weeks, Day 20 values of calcium precipitates were lower than in controls, but Day 20 ALP values were paradoxically higher. The correlation between calcium and ALP within each group was low. The correlation slightly improved, in uninhibited cultures, when Day 21 calcium values were matched with earlier Day 12 ALP values. This suggested the existence of a Levamisole-sensitive mechanism for mineralization inhibition effective prior to the culture's mineralization stage. To focus on this early effect on mineralization Levamisole was added to stromal cultures on different days and removed on Day 12. Levamisole decreased Day 21 mineralization when added on Days 0, 3, 5, and 7, but not when added on Day 9. The Levamisole-induced inhibition of mineralization was accompanied by an increase in Day 12 ALP specific activity, compared to controls, when added from Day 5 and thereafter. The results indicate that part of the ability of stromal cells to mineralize is determined during the first week of culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227185 TI - Murine osteoblast interleukin 4 receptor expression: upregulation by 1,25 dihydroxyvitamin D3. AB - The immune cytokine interleukin 4 has newly recognized effects on skeletal metabolism. While the interaction of many cells ultimately determines bone mass, we have examined the possibility that the osteoblast may be an IL-4 target in bone by characterizing IL-4 receptor (IL-4R) expression by MC3T3-E1 (MC3T3) murine osteoblastic cells. Based on 125I-IL-4 binding, MC3T3 cells express large numbers of IL-4 receptors (125I-IL-4 Bmax = 3,000-7,500 sites/cell, 125I-IL-4 K = 13-40 pM) with an affinity similar to the IL-4 receptor expressed by an IL-4 responsive T cell line. Monoclonal anti-IL-4R antibodies (M1) blocked specific MC3T3 125I-IL-4 binding and MC3T3 total cell RNA contained full-length IL-4R mRNA as detected by reverse transcription DNA amplification utilizing IL-4R primers and Northern blot analysis. Functionally, IL-4 treatment of MC3T3 cells resulted in increased cellular proliferation (10-20%) and inhibition of alkaline phosphatase levels (20-40%). While parathyroid hormone (PTH) exposure did not influence IL-4R levels, vitamin D3 treatment augmented MC3T3 125I-IL-4 binding, in a time-dependent manner, up to threefold after a 24 h exposure with a metabolite specificity indicating the involvement of the vitamin D receptor. Equilibrium binding studies showed that the impact of 1,25 (OH)2 D3 on MC3T3 125I IL-4 binding was due to an increased IL-4R Bmax. Cycloheximide treatment inhibited 1,25 (OH)2 D3-induced IL-4R upregulation, suggesting that protein synthesis was required. Furthermore, the steroid increased steady-state IL-4R mRNA levels in both a time- and concentration-dependent manner. The IL-4R message half-life was not altered by 1,25 (OH)2 D3, suggesting that increased IL-4R mRNA expression resulted from increased IL-4R gene transcription. Taken together, these findings raise the possibility that IL-4's influence on mineral metabolism could be mediated by osteoblasts and that the effectiveness of this cytokine may be influenced by vitamin D3's impact on IL-4R expression. PMID- 8227186 TI - Expression and localization of elements of the plasminogen activation system in benign breast disease and breast cancers. AB - The malignant potential of solid tumors is related to the ability to invade adjacent tissue and to metastasize. These properties of cancer cells depend on the synthesis of proteolytic enzymes which are able to digest adjacent connective tissue and basement membranes. We hypothesized that all elements of the plasminogen activation system might be overexpressed in malignant human breast tumors, functioning as an essential element in tumor invasion and metastasis. As determined by histopathological methods, the malignant tumors showed statistically significantly higher expression of urokinase plasminogen activator (uPA), type-1 plasminogen activator inhibitor (PAI-1), and especially urokinase plasminogen activator receptor (uPAR) than benign tissues. All those elements were present in higher amounts in the cancer cells than in the cells of benign or normal breast tissues. High exhibition of tissue plasminogen activator (tPA) found in cancer seems to be random and not related to the malignant or benign state, since benign and malignant tumors show overexpression of tissue plasminogen activator with similar frequency. When the tumors express high amounts of uPA, they express a high amount of uPAR in 50% of cases and PAI-1 in 57.3% of cases. When urokinase is expressed in low amount, the receptor is low in 28.6% and inhibitor in 21.4% of malignant breast tumors. This statistically significant consensus, 78.6% in the case of urokinase and its receptor and 78.6% in case of urokinase and its inhibitor, suggests that these activities may be the result of a unique mechanism of control, activated in the last steps of malignant transformation. PMID- 8227187 TI - 1,25-Dihydroxyvitamin D3 and macrophage colony-stimulating factor-1 synergistically phosphorylate talin. AB - Macrophage colony stimulating factor (CSF-1) and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) are potent inducers of macrophage differentiation. Both appear to modulate protein phosphorylation, at least in part, through protein kinase C (PKC) raising the question as to whether they concurrently impact on macrophage like cells. In this regard, we utilized the CSF-1 dependent murine macrophage like line BAC 1.25F5. CSF-1 treatment of these cells for 30 min leads to particular phosphorylation of a 165 kDa protein, the putative CSF-1 receptor, and a 210 kDa moiety. 1,25(OH)2D3 exposure for 24 h prior to addition of CSF-1 enhances phosphorylation of the 165 kDa species and, especially, the 210 kDa protein. Phosphorylation of the latter protein is 1,25(OH)2D3 dose- and time dependent and the molecule is specifically immunoprecipitated with a rabbit polyclonal anti-talin antibody. Experiments with okadaic acid show that the enhanced phosphorylation of talin does not result from serine phosphatase inhibition. CSF-1 and 1,25(OH)2D3, alone or in combination, do not increase talin protein expression. The tyrosine kinase inhibitor, genestein, blocks 1,25(OH)2D3/CSF-1 induced phosphorylation of the putative CSF-1 receptor but has no effect on talin phosphorylation which occurs exclusively on serine. In contrast to genestein, staurosporin, an inhibitor of PKC, inhibits phosphorylation of talin. Moreover, exposure of 1,25(OH)2D3 pretreated cells to phorbol 12-myristate 13-acetate (PMA) in place of CSF-1 also prompts talin phosphorylation. Finally, 1,25(OH)2D3 enhances 3[H]PDBu binding, indicating that the steroid increases PMA receptor capacity. Thus, CSF-1 and 1,25(OH)2D3 act synergistically via PKC to phosphorylate talin, a cytoskeletal-associated protein. PMID- 8227188 TI - Histone H1t: a tissue-specific model used to study transcriptional control and nuclear function during cellular differentiation. AB - One of the most prominent and best studied family of genes is the histone gene family. In recent years, histone gene regulation during the cell cycle of somatic cells has been studied extensively. This paper is intended to highlight and emphasize recent data concerning the tissue-specific expression of histone H1t using spermatogenesis as a model system. In this article we describe a unique DNA element within the proximal promoter of the histone H1t gene. This element has been shown to bind exclusively to nuclear proteins from pachytene spermatocytes and early spermatids. Thus, there is a strong temporal correlation between the appearance of the testis-specific DNA-binding protein and the onset of transcription of the testis-specific histone H1t gene. PMID- 8227189 TI - Vasopressin induces arachidonic acid release through pertussis toxin-sensitive GTP-binding protein in aortic smooth muscle cells: independence from phosphoinositide hydrolysis. AB - We previously reported that pertussis toxin (PTX) had little effect on arginine vasopressin-induced formation of inositol trisphosphate (IP3) in rat aortic smooth muscle cells [Kondo et al.: Biochemical and Biophysical Research Communications 161:677-682, 1989]. In the present study, we investigated the mechanism of vasopressin-induced arachidonic acid release in rat aortic smooth muscle cells. Vasopressin stimulated both the release of arachidonic acid and the formation of IP3 dose dependently in the range between 10 pM and 1 microM. The effect of vasopressin on arachidonic acid release was more potent than that on the formation of IP3. Quinacrine, a phospholipase A2 inhibitor, significantly suppressed the vasopressin-induced arachidonic acid release but had little effect on the formation of inositol phosphates. NaF, a GTP-binding protein activator, mimicked vasopressin by stimulating the arachidonic acid release. The arachidonic acid release stimulated by a combination of vasopressin and NaF was not additive. PTX partially but significantly suppressed the vasopressin-induced arachidonic acid release. In the cell membranes, PTX catalyzed ADP-ribosylation of a protein with an M(r) of about 40,000. Pretreatment of membranes with 0.1 microM vasopressin in the presence of 2.5 mM MgCl2 and 100 microM GTP markedly attenuated this PTX-catalyzed ADP-ribosylation of the protein in a time-dependent manner. These results strongly suggest that PTX-sensitive GTP-binding protein is involved in the coupling of vasopressin receptor to phospholipase A2 in primary cultured rat aortic smooth muscle cells. PMID- 8227190 TI - Endothelin modulates osteopontin and osteocalcin messenger ribonucleic acid expression in rat osteoblastic osteosarcoma cells. AB - Endothelins (ETs) are vasoconstrictive peptides produced mainly by endothelial cells. The ET receptors are expressed in many types of cells including osteoblast like cells. The purpose of this study was to examine the effects of endothelin on the expression of osteoblastic phenotype-related genes. We found that endothelin 1 (ET-1) enhanced approximately two-fold the mRNA expression of both osteopontin and osteocalcin genes in rat osteoblastic osteosarcoma ROS17/2.8 cells. These effects were dose-dependent, peaking at 10(-7) M. The ET-1 enhancement of the abundance of osteopontin and osteocalcin mRNAs was time-dependent, with a maximal effect at 24 h. ET-1 modulation of the expression of the two phenotype-related gene products of osteoblasts suggests that endothelin is one of the cytokines which modulate osteoblastic functions and that this molecule may play a role in the regulation of bone metabolism. PMID- 8227191 TI - Antibodies specific for gp40 inhibit cell-cell adhesion by cross-linking the protein on the surface of Dictyostelium purpureum. AB - We have previously suggested a role for gp40 in cell-cell adhesion in Dictyostelium purpureum from the fact that antibodies specific for this protein inhibited adhesion in an in vitro assay [Springer: Dev Biol 133:447-455, 1989]. To further confirm the role mutants lacking the protein were isolated and characterized. To our surprise, the mutants had normal adhesive properties both in vitro and in situ. These results lead us to the conclusion that gp40 is not necessary for the cell-cell adhesions observed and may not be a molecule which directly participates in these adhesions. When studied further, we found that adhesion-inhibitory antibodies were only effective as divalent IgG. Monovalent Fab fragments of the same antibodies could not inhibit adhesion. The inhibitory antibodies also caused the cells to remain rounded and incapable of attaching to plastic surfaces. We conclude that when divalent antibodies specific for gp40 cross-link this protein on the cell surface a cytoskeletal change prevents them from attaching to substratum as well as to other cells, thereby inhibiting cell cell adhesion. We suggest that an alternative mechanism for inhibition of cell cell adhesion by divalent antibodies exists and should be considered before proposing a direct role for a protein in adhesion. PMID- 8227192 TI - Isolation and characterization of a new class of acidic glycans implicated in sea urchin embryonal cell adhesion. AB - Three major glycan fractions of 580 kDa (g580), 150 kDa (g150), and 2 kDa (g2) were isolated and purified from Lytechinus pictus sea urchin embryos at the mesenchyme blastula stage by gel filtration and high pressure liquid chromatography. Chemical analysis, by gas chromatography, revealed that g580 is highly sulfated and rich in N-acetylglucosamine, N-acetylgalactosamine, glucuronic acid, and fucose. The g150 fraction is less acidic than g580 and contains high amounts of amino sugars, xylose, and mannose. The g2 fraction is neutral, rich in N-acetylglucosamine, mannose, and galactose. The g580 and g150 fractions are resistant to glycosaminoglycan-degrading enzymes, indicating that they are distinct from the glycosaminoglycans. The g580 fraction resembles, with respect to chemical composition, a previously characterized 200 kDa sponge adhesion glycan (g200). The binding of the monoclonal antibody Block 2, which recognizes a repetitive epitope on g200, as well as of the anti-g580 polyclonal antibodies to both g580 and g200 indicated that these two glycans share similar antigenic determinants. The Fab fragments of the Block 2 antibody, which previously have been shown to inhibit cell adhesion in sponges, also blocked the reaggregation of dissociated sea urchin mesenchyme blastula cells. These results indicate that g580 carries a carbohydrate epitope, similar to the sponge adhesion epitope of g200, which is involved in sea urchin embryonal cell adhesion. PMID- 8227193 TI - Ezrin is concentrated in the apical microvilli of a wide variety of epithelial cells whereas moesin is found primarily in endothelial cells. AB - Ezrin and moesin are two cytoskeletal proteins originally purified from human placenta that are 74% identical in overall protein sequence. They are believed to be membrane-cytoskeletal linking proteins because they share sequence homology with erythrocyte band 4.1 and colocalize with actin specifically in microvilli and membrane ruffles in cultured cells. To determine if ezrin and moesin share similar distributions in vivo, we studied their localizations with respect to F actin in tissue sections. Surprisingly, ezrin and moesin exhibited very different cellular distributions. Ezrin was highly concentrated and colocalized with actin on the apical surface of many epithelial cell types. During enterocyte differentiation, the pattern of expression and redistribution of ezrin was consistent with it performing a role in microvillus assembly. Immunoelectron microscopy in differentiated cells revealed that ezrin was restricted mainly to the plasma membrane of microvilli and other actin-rich surface projections. Moesin was found in endothelial cells and was also enriched in the apical microvilli of a restricted set of epithelial cells. All polarized cell types with abundant microvilli contained one or both proteins, suggesting that ezrin and moesin perform related functions. However, the differential expression of ezrin and moesin indicates that they have distinct properties, which are uniquely adapted to specific cell types. PMID- 8227194 TI - Chick corneal development in vitro: diverse effects of pH on collagen assembly. AB - In vivo, the embryonic chick corneal epithelium lays down a stroma of collagen and proteoglycans whose fibrils are unusual as their diameter distribution peaks sharply about a mean of 20 nm. Such epithelia cultured on Nuclepore filters will also lay down a stroma containing 20 nm diameter fibrils, although there is only limited orthogonal organisation. We report here that collagen fibril morphology is critically dependent on the pH of the medium in which the corneal epithelium is cultured and that normal 20 nm diameter fibrils only assemble in a narrow band around neutral pH (approx. 6.9-7.4). At higher pH (7.6-8.1), fibrils in the distal region of the stroma more closely resemble those seen in non-corneal stroma as their diameters can be up to 200 nm even though fibrils near the basal lamina are only about 10 nm in diameter. At low pH (approx. 6.5), there are again wide fibrils, but with the hieroglyphic cross-sections typical of those seen in heritable disorders of N-terminal procollagen processing. Biochemical analysis by SDS-PAGE and fluorography confirms that N-terminal procollagen processing is deficient at this pH. At very low pH (approx. 5.8-6.2), there is little processing of procollagen and the stroma comprises filamentous material with the occasional banded structures typical of those formed by unprocessed procollagen at high concentration. Gel electrophoresis and peptide mapping showed that the collagens produced by the corneal epithelium of the primary stroma included types I, II and V and that total collagen production, as assessed by incorporation of [3H]proline, increased with pH, although the relative amounts of the different collagens produced remained essentially unchanged. While the biochemical data can account for the altered morphologies in the pH range 5.8 to 7.0, the sensitivity of fibril diameter to small changes around neutral pH remains unexplained, but points to the subtle, charge-based interactions necessary for the formation of 20 nm diameter fibrils in the developing cornea. PMID- 8227195 TI - Reversion analysis of dynein intermediate chain function. AB - The ODA6 locus of Chlamydomonas reinhardtii encodes a 70 kDa intermediate chain protein of the flagellar outer row dynein ATPase, and mutations at this locus prevent assembly of the entire outer row dynein arm complex. To initiate a structure-function analysis of the 70 kDa protein, we used transformation with chimeric mutant/wild-type genes to localize the defect in one assembly mutation, oda6-95. Sequence analysis revealed a frame-shift mutation in codon 53, which is followed by a stop codon after 13 amino acids in the new reading frame. By selecting intragenic pseudorevertants of this mutation we obtained 11 new oda6 alleles. Many of these pseudorevertants encode intermediate chain proteins that permit assembly of outer row arms but do not restore full wild-type motility. Revertant strains fall into two phenotypic classes, one with average beat frequencies of 54 Hz (similar to wild type) and one with average frequencies of 27 Hz (compared with 24 Hz for oda6-95) during normal forward swimming. Low beat frequency strains also display abnormalities during photophobic reversal (symmetric waveform). Amplification and sequence analysis of revertant alleles indicated that each reversion caused a second frame-shift, within a 115 nt interval, which restored the original reading frame, and that phenotypic severity was related to both direction (5' or 3') and distance between the original mutation and the reversion event. On the basis of immunoblot analysis of outer arm proteins, we conclude that revertant motility defects do not correlate with deficits in assembly of a specific dynein heavy chain or intermediate chain polypeptide, and electron microscopy confirms that revertants have normal outer arm structures. These results suggest that the 70 kDa intermediate chain plays a direct role in outer arm function distinct from its role in the assembly process. PMID- 8227196 TI - Alpha melanocyte stimulating hormone (alpha MSH) stimulates normal human melanocyte growth by binding to high-affinity receptors. AB - The combined action of cholera toxin (CT)-dependent activation of the adenylate cyclase signaling pathway, stimulation of protein kinase C, and activation of the tyrosine kinase activity of cell surface receptors and proto-oncogene products, have been shown to stimulate melanocyte proliferation. However, natural factors responsible for the optimal stimulation of normal human melanocyte growth, either isolated or co-cultured with keratinocytes, remain largely unknown. alpha MSH (alpha melanocyte stimulating hormone) has previously been shown to bind to murine and human melanoma cells and to stimulate their adenylate cyclase and tyrosinase activity. In contrast, very little is known about the presence and function of alpha MSH receptors in normal human melanocytes. We now report that alpha MSH: (i) binds to normal human melanocytes through a single class of high affinity receptors; (ii) does not induce per se melanocytes to enter the S-phase of the cell cycle; (iii) does indeed stimulate melanocyte proliferation in a dose dependent fashion; but its stimulatory effect requires bFGF and/or the activation of protein kinase C. PMID- 8227197 TI - Heparan sulfate proteoglycan and FGF receptor target basic FGF to different intracellular destinations. AB - Basic FGF is a prototype of a family of heparin binding growth factors that regulate a variety of cellular responses including cell growth, morphogenesis and differentiation. At least two families of receptors bind bFGF and could mediate its response: (1) tyrosine kinase-containing FGF receptors, designated FGFR-1 to FGFR-4, and (2) heparan sulfate proteoglycans that bind bFGF through their heparan sulfate chains. Both are known to undergo internalization and thus bFGF bound to the different receptors may be internalized via more than one pathway. It is not known whether the intracellular fate of bFGF differs depending upon which receptor binds it at the cell surface. To investigate the respective roles of these receptors in the intracellular targeting of bFGF, we utilized NMuMG cells that bind and internalize bFGF through their heparan sulfate proteoglycans, but do not express detectable levels of FGFRs nor respond to bFGF. Basic FGF conjugated to saporin (bFGF-saporin) was used as a probe to study targeting of bFGF by the different receptors. Saporin is a cytotoxin that has no effect on cells if added exogenously. However, it kills cells if it gains access to the cytoplasm. The NMuMG cells internalize bFGF-saporin but are not killed. Transfecting these cells with FGFR-1 results in bFGF-responsive cells, which bind and internalize bFGF through FGFR-1, and are killed. Removing the heparan sulfate from these cells eliminates killing by bFGF-saporin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227198 TI - Six git genes encode a glucose-induced adenylate cyclase activation pathway in the fission yeast Schizosaccharomyces pombe. AB - An important eukaryotic signal transduction pathway involves the regulation of the effector enzyme adenylate cyclase, which produces the second messenger, cAMP. Previous genetic analyses demonstrated that glucose repression of transcription of the Schizosaccharomyces pombe fbp1 gene requires the function of adenylate cyclase, encoded by the git2 gene. As mutations in git2 and in six additional git genes are suppressed by exogenous cAMP, these 'upstream' git genes were proposed to act to produce a glucose-induced cAMP signal. We report here that assays of cAMP levels in wild-type and various mutant S. pombe cells, before and after exposure to glucose, show that this is the case. The data suggest that the cAMP signal results from the activation of adenylate cyclase. Therefore these 'upstream' git genes appear to encode a glucose-induced adenylate cyclase activation pathway. Assays of cAMP on a strain carrying a mutation in the git6 gene, which acts downstream of adenylate cyclase, indicate that git6 may function to feedback regulate adenylate cyclase activity. Thus git6 may encode a cAMP dependent protein kinase. PMID- 8227199 TI - Molecular cloning and cellular localization of a BiP homologue in Trypanosoma brucei. Divergent ER retention signals in a lower eukaryote. AB - Using the polymerase chain reaction with degenerate primers, three new members of the hsp70 gene family of Trypanosoma brucei have been identified. A genomic clone of one of these, gA, has been fully sequenced and the corresponding gene product has been characterized using antibody to recombinant gA fusion protein. gA is the trypanosomal homologue of BiP, an endoplasmic reticulum resident hsp70 gene family member, based on four lines of evidence: (1) gA protein has 64% deduced amino acid identity with rat BiP; (2) the deduced amino acid sequence has a putative secretory signal peptide; (3) the gA gene product is a soluble luminal resident of a trypanosomal microsome fraction; (4) the gA polypeptide does not cofractionate with mitochondrial markers. Trypanosomes are the most primitive eukaryote yet in which BiP has been identified. The gA polypeptide has been used as a specific marker for the direct visualization of endoplasmic reticulum in trypanosomes by both indirect immunofluorescence and cryoimmuno electron microscopy. The endoplasmic reticulum is seen as a tubular network that extends throughout the cell excluding the flagellum. The C-terminal tetrapeptide of gA is MDDL, which, together with the C-terminal tetrapeptide (KQDL) of a trypanosome protein disulfide isomerase homologue (Hsu et al. (1989) Biochemistry 28, 6440 6446), indicates that endoplasmic reticulum retrieval signals in trypanosomes may be as divergent and heterogeneous as any seen in the other eukaryotes yet studied. PMID- 8227200 TI - The BiP protein and the endoplasmic reticulum of Schizosaccharomyces pombe: fate of the nuclear envelope during cell division. AB - A polyclonal antibody was raised to the C-terminal region of fission yeast BiP. The use of this antibody for immunoprecipitation, western blotting and immunofluorescence has confirmed and extended the observations made previously with an epitope-tagged BiP molecule. A fraction of BiP protein is glycosylated in Schizosaccharomyces pombe cells. Pulse-chase experiments showed that this modification occurs rapidly upon synthesis and that the extent of glycosylation does not then change with time. BiP protein is induced by elevated temperatures and by treatment with tunicamycin. The antibody cross-reacts with proteins of similar molecular weight in the yeasts Kluyveromyces lactis and Schizosaccharomyces japonicus. Immunofluorescence of BiP has been used to follow the behaviour of the ER and in particular the nuclear envelope through the cell cycle. PMID- 8227201 TI - Evidence for two intracellular calcium pools in Dictyostelium: the cAMP-induced calcium influx is directed into a NBD-Cl- and 2,5-di-(tert-butyl)1,4-hydroquinone sensitive pool. AB - Signal transduction in Dictyostelium for oriented movement and differentiation involves a fine tuning of the cytosolic Ca2+ concentration. We have previously shown that cAMP binding to the cell surface receptor elicits two cellular events: (i) to enhance Ca2+ entry across the plasma membrane; (ii) to increase Ca2+ uptake into Ca(2+)-sequestering organelles. Here we used permeabilised cells to show that cAMP-induced Ca2+ uptake in these cells was sensitive to the Ca2+ transport ATPase blocker 2,5-di-(tert-butyl)-1,4-hydroquinone (BHQ) and the vacuolar H(+)-ATPase inhibitor NBD-Cl. By contrast, bafilomycin A1 and vanadate, inhibitors of Ca2+ uptake into acidosomes in Dictyostelium, did not reduce the cAMP-induced Ca2+ uptake of permeabilised cells. GTP gamma S served as a tool to measure Ins(1,4,5)P3- (InsP3)-sensitive Ca2+ release. Following NBD-Cl or BHQ treatment Ca2+ release was reversibly inhibited. We conclude that the cAMP controlled Ca2+ influx is directed into a NBD-Cl and BHQ-sensitive compartment, which comprises the InsP3-releasable pool. The acidosomal Ca2+ store seems to provide for additional Ca2+ if required. PMID- 8227202 TI - Association of the brain anion exchanger, AE3, with the repeat domain of ankyrin. AB - The 89 kDa NH2-terminal domain of erythrocyte ankyrin is composed almost entirely of 22 tandem repeats of a 33 amino acid sequence and constitutes the binding site for the cytoplasmic NH2-terminal domain of the erythrocyte anion exchanger, AE1. We have developed an assay to evaluate the in vivo interaction between a fragment of ankyrin corresponding to this domain (ANK90) and two non-erythroid anion exchangers, AE2 and AE3, that share considerable structural homology with AE1. Association was assessed by co-immunoprecipitation of ANK90-anion exchanger complexes from detergent extracts of cells cotransfected with plasmids encoding the ankyrin fragment and the anion exchanger or mutants thereof. ANK90 was co immunoprecipitated with AE1 but not with an AE1 deletion mutant lacking the cytoplasmic NH2-terminal domain. Using this assay, we show that the brain anion exchanger AE3, but not the closely related homologue, AE2, is capable of binding to ankyrin. PMID- 8227203 TI - Transcriptionally active minichromosomes are attached transiently in nuclei through transcription units. AB - Viral minichromosomes provide simple models for chromatin domains. The sequences attaching them to larger nuclear structures were mapped; attachments were defined operationally by their ability to prevent chromatin fragments electroeluting from nuclei. Cells encapsulated in agarose microbeads were lysed and analysed subsequently in a 'physiological' buffer. Non-transcribed minichromosomes eluted from nuclei but transcriptionally active ones did not. Cutting attached minichromosomes with HaeIII enabled most of the resulting approximately 400 bp fragments to elute. Analysis of residual fragments showed that no single sequence was responsible for attachment; rather, minichromosomes were attached at only one or two points through a promoter or part of a transcription unit. This suggests that RNA polymerases mediate attachments that change dynamically as the template slides past the attached enzyme. As, under optimal conditions, polymerases continue to elongate roughly at the rate found in vivo, these attachments are unlikely to be generated artifactually. PMID- 8227204 TI - Localization of 5 S genes and transcripts in Pisum sativum nuclei. AB - We have investigated the position of the 5 S gene clusters in the nuclei of Pisum sativum root tip cells, and of their transcripts in the nucleolus, using in situ hybridization and confocal microscopy. Single-stranded RNA probes were produced by in vitro transcription, incorporating fluorescein or digoxygenin label. There are known to be 3 pairs of 5 S gene clusters in this species, which would be expected to give 6 sites of hybridization in G1 cells and 12 sites in G2 cells. In presumed G2 cells, many of the sites appeared as paired spots very close together, which we interpret as the sites on sister chromatids. A clear association of one or more clusters with the nucleolar periphery was observed, and it is possible that this has a functional significance. The transcript labelling within the nucleolus showed a reproducible and highly ordered pattern, consisting of cavities devoid of transcript labelling surrounded by sheets of bright labelling, making a characteristic and often very symmetrical cage-work structure. This labelling pattern may represent an underlying organization of transcript processing within the nucleolus, and is highly reminiscent of the distribution of a previously described nucleolar matrix protein. PMID- 8227205 TI - Detection of DNA in the nucleoids of chloroplasts and mitochondria in Euglena gracilis by immunoelectron microscopy. AB - DNA in the nucleoids of chloroplasts and of mitochondria in Euglena gracilis was detected with anti-DNA antibodies by immunoelectron microscopy. After treatment with the antibodies, DNA in these organelles combined with gold particles that had been coated with anti-IgM antibodies such that it was possible to trace the outlines of the nucleoids. Nucleoids in chloroplasts appeared to be composed of twisted threads 50-70 nm in diameter. The twisted threads were entangled to form thicker nodes of 100-200 nm diameter. Most nucleoids in mitochondria were spherical or ovoid, 70-130 nm in diameter. Nucleoids both in chloroplasts and in mitochondria contained cores with which DNA threads were in tight contact. The structure of the nucleoids was very different from those previously observed by conventional electron microscopy. PMID- 8227206 TI - The tumor promoter 12-O-tetradecanoylphorbol-13-acetate blocks differentiation of HT-29 human colon cancer cells. AB - Recently developed HT-29-derived cell lines, which display variable differentiated phenotypes provide an invaluable opportunity to analyze the mechanism by which cell differentiation is regulated in the intestine. We have studied the effects of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) in the differentiation phenotype of mucus-secreting (HT-29 M6) and absorptive (HT-29 M3) cells. TPA prevented the accumulation of differentiation markers such as dipeptidylpeptidase IV, villin or mucins, down-regulated the expression of these molecules in post-confluent differentiated cell cultures and induced the loss of the functional integrity of the tight junction in the monolayer (i.e. decreased transepithelial resistance and inhibited dome formation). These effects were mediated by activation of protein kinase C (PK-C), as demonstrated using the specific inhibitor GF109203x. Analysis by immunoblotting of the PK-C isoforms present in HT-29 M6 cells revealed that the most abundant TPA-sensitive isoform was PK-C epsilon, although low levels of cPK C were also detected. Further studies are necessary to elucidate the role of the different PK-C isoforms in the differentiation of HT-29 cells. PMID- 8227207 TI - Cell-to-cell transport via motile tubules in growing hyphae of a fungus. AB - The system of pleiomorphic, motile tubules and vacuoles in growing hyphal tips of Pisolithus tinctorius has been shown to play a role in intracellular transport. Here we show that the same system also exchanges material between adjacent cells. This exchange is most obvious between terminal and penultimate cells following nuclear division in the tip cell and just before dissolution of the cell wall between the clamp connection and penultimate cell. At this stage the two new dolipore septa are complete. The process was studied in living hyphae using confocal and conventional fluorescence microscopy. Tubules could move in either direction across the septum and often extended and retracted several times and penetrated for some distance (e.g. 40 microns) into the receiving cell. Movements appeared co-ordinated and during the exchange tubules transiently interconnected vacuoles in adjacent cells and by peristaltic movements appeared to transfer material between them. The fluorescent tubules occupied a specific plane in the vicinity of the septum and remained in this plane for the duration of their movement, suggesting that their orientation and direction of movement is controlled. In freeze-substituted hyphae, tubular cisternae of similar dimensions to fluorescent tubules passed through the parenthesome pores perpendicular to the septum and in some cases entered the mouth of the septal pore. This indicates that the septal pore is of an appropriate dimension to accommodate the tubules and that they can cross the septal pore to exchange material between vacuole systems of adjacent cells. This is the first direct demonstration of such intercellular transport via a sub-cellular compartment. PMID- 8227208 TI - Cotranslational assembly of some cytoskeletal proteins: implications and prospects. PMID- 8227209 TI - Association of p34cdc2/cyclin B complex with microtubules in starfish oocytes. AB - The microtubular cytoskeleton exhibits a dramatic reorganization, progressing from interphase radial arrays to a mitotic spindle at the G2/M transition. Although this reorganization has been suspected to be caused by maturation promoting factor (MPF: p34cdc2/cyclin B complex), little is known about how p34cdc2 kinase controls microtubule networks. We provide evidence of the direct association of the p34cdc2/cyclin B complex with microtubules in starfish oocytes. Anti-cyclin B staining of detergent-treated oocytes, isolated asters and meiotic spindles revealed fluorescence associated with microtubule fibers, chromosomes and centrosomes. Microtubules prepared from starfish oocytes were associated with cyclin B and p34cdc2 proteins. Microtubule-bound p34cdc2 and cyclin B were released from microtubules by a high-salt solution and possessed a complex form as shown by the adsorption to suc1-beads and by immunoprecipitation with the anti-cyclin B antibody. The p34cdc2/cyclin B complex associated to microtubules had high histone H1 kinase activity at meiotic metaphase. However, it was not necessary for the p34cdc2/cyclin B complex to be active for microtubule binding, as an inactive form in immature oocytes was also observed to bind to microtubules. The coprecipitation of suc1-column purified p34cdc2/cyclin B with purified porcine brain microtubules in the presence of starfish oocyte microtubule-associated proteins (MAPs) indicates that the association of p34cdc2/cyclin B with microtubules in vitro is mediated by MAPs. PMID- 8227210 TI - Changes in the cytoskeletal structure of cultured smooth muscle cells induced by calyculin-A. AB - Changes in the cytoskeletal structure of cultured A10 smooth muscle cells induced by calyculin-A (CL-A), a potent inhibitor of types 1 and 2A protein phosphatases, were analyzed using indirect fluorescence techniques. In the presence of 1 x 10( 7) M CL-A the cells became round and subsequently detached from the substratum. The effect of CL-A was inhibited by a non-selective kinase inhibitor, K-252a, but not by EGTA. In rounded cells stress fibers were absent and staining for F-actin appeared in patches. Vinculin, one of the components of focal contacts, was localized at the periphery of control cells. CL-A treatment moved the focal contacts towards the inside of the cell along the stress fibers, and this was followed by the rounding up of the cell. In addition, rapid and marked changes in microtubule structure were observed in CL-A-treated cells. Many 'nicks' or 'gaps' were observed along the microtubules in the attached, spread cells. A filamentous network of microtubules was not observed in the detached cells, i.e. after longer exposure to CL-A. These results suggest that CL-A may change the structure of focal contacts, resulting in the rounding up of the cell, and inducing a microtubule-severing activity. These effects were independent of the external Ca2+ concentration. The changes in cytoskeletal structure may be caused by disturbing the balance of phosphorylation and dephosphorylation in the cell. PMID- 8227211 TI - Identification and preliminary characterization of a 65 kDa higher-plant microtubule-associated protein. AB - Microtubules in plant cells, as in animal cells, are dynamic structures. However, our lack of knowledge about the constituents of microtubules in plant cells has prevented us from understanding the mechanisms that control microtubule dynamics. To characterize some of these constituents, a cytoplasmic extract was prepared from evacuolated protoplasts (miniprotoplasts) of tobacco BY-2 cells, and microtubules were assembled in the presence of taxol and disassembled by cold treatment in the presence of Ca2+ and a high concentration of NaCl. SDS-PAGE analysis of triple-cycled microtubule protein revealed the presence of 120 kDa, 110 kDa and a group of 60-65 kDa polypeptides in addition to tubulin. Since these polypeptides had copolymerized with tubulin, through the three cycles of assembly and disassembly, and they bundle microtubules, we tentatively identified the three polypeptides as microtubule-associated proteins (MAPs). To characterize these factors further, triple-cycled microtubule protein was fractionated by Mono Q anion-exchange chromatography and the microtubule-bundling activity of each fraction was examined. Fractions having microtubule-bundling activity contained only the 65 kDa MAP, an indication that the 65 kDa MAP is responsible for the bundling of microtubules. Purified 65 kDa MAP formed cross-bridge structures between adjacent microtubules in vitro. Polyclonal antibodies were raised in mice against the 65 kDa MAP. Immunofluorescence microscopy revealed that the 65 kDa MAP colocalized with microtubules in BY-2 cells throughout the cell cycle. Western blotting analysis of extracts from several species of plants suggested that the 65 kDa MAP and/or related peptides are widely distributed in the plant kingdom. PMID- 8227212 TI - The highly divergent alpha- and beta-tubulins from Dictyostelium discoideum are encoded by single genes. AB - As a step in the characterization of the microtubule system of Dictyostelium discoideum, we have isolated and sequenced full-length cDNA clones that encode the Dictyostelium alpha- and beta-tubulins, as well as the Dictyostelium alpha tubulin gene. Southern blot analysis suggests that Dictyostelium is unusual in that its genome contains single alpha- and beta-tubulin genes, rather than the multi-gene family common in most eukaryotic organisms. The complete alpha-tubulin cDNA contains 1558 nucleotides, with an open reading frame, that encode a protein of 457 amino acids. The complete beta-tubulin cDNA contains 1572 nucleotides and encodes a protein of 456 amino acids. Analysis of the deduced protein sequences indicates that while there is a significant degree of sequence similarity between the Dictyostelium tubulins and other known tubulins, the Dictyostelium alpha tubulin displays the greatest sequence divergence yet described. Single alpha- and beta-tubulin transcripts are detected by northern blot analysis during all stages of Dictyostelium development. The highest levels of message accumulate late in germinating spores and vegetative amoebae. Despite changes in alpha- and beta-tubulin mRNA levels, protein levels remain constant throughout development. We have expressed the carboxy-terminal two-thirds of the alpha- and beta-tubulins as trpE fusions in Escherichia coli and used this protein to produce polyclonal antisera specific for the Dictyostelium alpha- and beta-tubulins. These antisera recognize one alpha- and two beta-tubulin spots on western blots of 2-D gels and, by indirect immunofluorescence, both recognize the interphase and mitotic microtubule arrays in vegetative amoebae. PMID- 8227213 TI - Spatial localization of agonist-induced Ca2+ entry in bovine adrenal chromaffin cells. Different patterns induced by histamine and angiotensin II, and relationship to catecholamine release. AB - The spatial organization of agonist-induced Ca2+ entry in single bovine adrenal chromaffin cells has been investigated using video-imaging techniques to visualize fura-2 quenching by the Ca2+ surrogate, Mn2+. The potent secretagogue histamine, in addition to releasing Ca2+ from intracellular stores, resulted in a large influx of external Mn2+ that occurred over the entire surface of the cell. The influx of Ca2+ that this mirrors was found to be an obligatory requirement for the triggering of catecholamine release by histamine, which suggests that such a global influx of Ca2+ into the cell probably underlies the ability of this agonist to stimulate a large secretory response. By contrast, the weaker secretagogue angiotensin II, which also acts through the second messenger inositol trisphosphate, produced a localized entry of external Mn2+ in 64% of cells. In these cells, localized Mn2+ entry always occurred at the pole of the cell in which the angiotensin II-induced rise in [Ca2+]i was largest. Since exocytosis in response to angiotensin II has previously been shown to be restricted to this same pole of the cell (Cheek et al. (1989). J. Cell Biol. 109, 1219-1227), these results suggest that localized influx of Ca2+ in response to angiotensin II could underlie the polarized exocytotic response observed with this stimulus. These results directly demonstrate that different agonists can induce different patterns of divalent cation influx in the same cells and, furthermore, suggest how these different patterns can have a direct influence on cellular function. PMID- 8227214 TI - Expression of E- or P-cadherin is not sufficient to modify the morphology and the tumorigenic behavior of murine spindle carcinoma cells. Possible involvement of plakoglobin. AB - Transfection of E- and P-cadherin cDNA has been carried out in murine spindle carcinoma cells previously shown to be deficient in both cadherins (Navarro et al., J. Cell Biol. 115, 517-533, 1991). High levels of expression of E- or P cadherin do not significantly affect the fibroblastic morphology of the parental spindle cells. In addition, the tumorigenic behavior of these highly malignant cells is not influenced by the ectopic expression of either cadherin. Nevertheless, a fraction of the exogenous cadherins is able to associate to detergent-insoluble components of the transfectant cells, and the expression of the exogenous E-cadherin confers Ca(2+)-dependent aggregation on the spindle transfectants in an in vitro assay. Immunoprecipitation analysis of the cadherin catenin complex of the transfectants revealed that the ectopic E-cadherin associates with the alpha- and beta-catenin proteins. However, the gamma catenin/plakoglobin component could not be detected in the E-cadherin immunocomplexes of the spindle transfectant cells, in contrast to the epithelial cells where the three catenins appeared to be associated with E-cadherin. The lack of association of gamma-catenin is correlated with very low levels of plakoglobin in whole cell extracts of the parental spindle cells. These results indicate that the association of E-cadherin with the alpha- and beta-catenin components is not sufficient to promote a fibroblastoid-epithelial conversion of highly malignant spindle cells. The presence of plakoglobin could be required for the proper organization of E-cadherin in the transfectant cells in order to acquire an epithelioid phenotype. PMID- 8227215 TI - The small GTP-binding protein, Rab6p, is associated with both Golgi and post Golgi synaptophysin-containing membranes during synaptogenesis of hypothalamic neurons in culture. AB - We have recently localized a small GTP-binding protein (Rab6p) thought to be involved in vesicular membrane transport, to the medial and trans-cisternae of the Golgi apparatus in NRK (normal rat kidney) cells. Here, we have localized and quantified Rab6p during the development in culture of embryonic neurons, up to synapse formation, and compared its subcellular distribution and level of expression to that of synaptophysin, a major integral membrane protein of small synaptic vesicles. Using immunocytochemistry (laser scanning confocal microscopy, immunoelectron microscopy), fractionation and immunoisolation methods, we show that during the early phase of synaptogenesis, Rab6p is associated with synaptophysin-containing membranes of a trans-Golgi subcompartment, post-Golgi vesicles and small synaptic vesicles or their precursors. Concomitantly, Rab6p undergoes translocation from cytosol to membranes and its level of expression increases. However, at late stages, the association of Rab6p to small synaptic vesicles sharply decreases and its level of expression plateaus. These findings suggest a role for Rab6p in the post-Golgi transport of synaptophysin, at an early step of the biogenesis of small synaptic vesicles. PMID- 8227216 TI - The control of chondrocyte differentiation during endochondral bone growth in vivo: changes in TGF-beta and the proto-oncogene c-myc. AB - The expression of transforming growth factor-beta and the c-myc proto-oncogene was studied in situ in the chondrocytes of the tibial growth plate of normal chicks and those with avian tibial dyschondroplasia in which the chondrocytes are developmentally arrested in the transitional phase between proliferation and differentiation. This results in an accumulation of unmineralised and avascular cartilage. Dyschondroplastic chicks showed reduced c-myc expression in the transitional chondrocytes but unaltered levels in the proliferating chondrocytes. Transforming growth factor-beta expression was reduced in the transitional chondrocytes of dyschondroplastic chicks. In areas where the lesion was being repaired there was evidence of increased expression of both c-myc protein and transforming growth factor-beta. Addition of 1,25-dihydroxyvitamin D to the diet, which is known to reduce the incidence of dyschondroplasia, resulted in an increase in c-myc production. These results suggest that both transforming growth factor-beta and the proto-oncogene c-myc may be important elements of the cascade of events that lead to chondrocyte differentiation, hypertrophy and mineralisation. PMID- 8227217 TI - Implanted myoblasts not only fuse with myofibers but also survive as muscle precursor cells. AB - Intramuscular implanted myoblasts can fuse with existing myofibers. Here we report that implanted primary myoblasts marked with retroviral transgenes can also persist as muscle precursor cells. These cells can be recovered as viable myoblasts from muscles of recipient mice even months after myoblast implantation, and they can fully resume expression of the transgenes in culture. Upon re implantation into muscles, they again not only fuse with existing myofibers, but also survive as muscle precursor cells in the tissue. These reserve myogenic cells should be able to contribute to host myofibers in muscle regeneration when the recombinant myofibers are damaged, providing an additional mechanism to maintain a persistent expression of transgenes delivered by myoblast-mediated gene transfer. PMID- 8227218 TI - Quantification and immunolocalisation of porcine articular and growth plate cartilage collagens. AB - The collagens of growth plate and articular cartilage from 5-6 month old commercial pigs were characterised. Growth plate cartilage was found to contain less total collagen than articular cartilage as a proportion of the dry weight. Collagen types I, II, VI, IX and XI are present in both growth plate and articular cartilage whereas type X is found exclusively in growth plate cartilage. Types III and V collagen could not be detected in either cartilage. Type I collagen makes up at least 10% of the collagenous component of both cartilages. There are significant differences in the ratios of the quantifiable collagen types between growth plate and articular cartilage. Collagen types I, II, and XI were less readily extracted from growth plate than from articular cartilage following pepsin treatment, although growth plate cartilage contains less of the mature collagen cross-links, hydroxylysyl-pyridinoline and lysyl pyridinoline. Both cartilages contain significant amounts of the divalent reducible collagen cross-links, hydroxylysyl-ketonorleucine and dehydro hydroxylysinonorleucine. Immunofluorescent localisation indicated that type I collagen is located predominantly at the surface of articular cartilage but is distributed throughout the matrix in growth plate. Types II and XI are located in the matrix of both cartilages whereas type IX is predominantly pericellular in the calcifying region of articular cartilage and the hypertrophic region of the growth plate. Collagen type VI is located primarily as a diffuse area at the articular surface. PMID- 8227219 TI - The spatial distribution and relative abundance of gap-junctional connexin40 and connexin43 correlate to functional properties of components of the cardiac atrioventricular conduction system. AB - Electrical coupling between heart muscle cells is mediated by specialised regions of sarcolemmal interaction termed gap junctions. In previous work, we have demonstrated that connexin42, a recently identified gap-junctional protein, is present in the specialised conduction tissues of the avian heart. In the present study, the spatial distribution of the mammalian homologue of this protein, connexin40, was examined using immunofluorescence, confocal scanning laser microscopy and quantitative digital image analysis in order to determine whether a parallel distribution occurs in rat. Connexin40 was detected by immunofluorescence in all main components of the atrioventricular conduction system including the atrioventricular node, atrioventricular bundle, and Purkinje fibres. Quantitation revealed that levels of connexin40 immunofluorescence increased along the axis of atrioventricular conduction, rising over 10-fold between atrioventricular node and atrioventricular bundle and a further 10-fold between atrioventricular bundle and Purkinje fibres. Connexin40 and connexin43, the principal gap-junctional protein of the mammalian heart, were co-localised within atrioventricular nodal tissues and Purkinje fibres. By applying a novel photobleach/double-labelling protocol, it was demonstrated that connexin40 and connexin43 are co-localised in precisely the same Purkinje fibre myocytes. A model, integrating data on the spatial distribution and relative abundance of connexin40 and connexin43 in the heart, proposes how myocyte-type-specific patterns of connexin isform expression account for the electrical continuity of cardiac atrioventricular conduction. PMID- 8227220 TI - The product of the Drosophila segment polarity gene armadillo is part of a multi protein complex resembling the vertebrate adherens junction. AB - Sequence similarity between the Drosophila segment polarity protein Armadillo and the vertebrate adherens junction protein beta-catenin raised the possibility that adherens junctions function in transduction of intercellular signals like that mediated by Wingless/Wnt-1. To substantiate the sequence similarity, properties of Armadillo were evaluated for consistency with a junctional role. Armadillo is part of a membrane-associated complex. This complex includes Armadillo, a glycoprotein similar in size to vertebrate cadherins, and the Drosophila homolog of alpha-catenin. Armadillo co-localizes with junctions that resemble vertebrate adherens junctions in morphology and position. These results suggest that Drosophila and vertebrate adherens junctions are structurally similar, validating use of Armadillo and its associated proteins as a model for vertebrate adherens junctions. PMID- 8227221 TI - Caring for the chronically ill. PMID- 8227222 TI - A model for the development and implementation of a patient support group in a medical-surgical setting. PMID- 8227223 TI - Animal-assisted therapy for the homebound elderly. PMID- 8227224 TI - Research program on learned response to chronic illness experience: self-help model. PMID- 8227225 TI - Caring for children with chronic conditions: model of critical times. PMID- 8227226 TI - Care of the chronically ill: paradigm shifts and directions for the future. PMID- 8227227 TI - Ethical framework for care of the chronically ill. PMID- 8227228 TI - Nursing's new political era. PMID- 8227229 TI - Growing up with a chronic condition: transition to young adulthood for the individual with cystic fibrosis. PMID- 8227230 TI - Project HOPE: developing health-care policy for host countries. PMID- 8227231 TI - Advances in pharmacotherapy: recent developments in the treatment of epilepsy. AB - Epilepsy is a disorder of the central nervous system in which the clinical symptoms are recurrent seizures. An increased understanding of the underlying mechanism of seizures and more definitive diagnostic procedures have improved the care of the patient with epilepsy. An improved classification of various seizure types, including specific epilepsy syndromes has helped optimize use of the standard antiepileptic drugs. Research on the mechanism of seizures has led to new antiepileptic drugs. More definitive diagnostic procedures have led to more accurate identification of patients likely to benefit from epilepsy surgery. This review focuses on these areas. PMID- 8227232 TI - Advances in pharmacotherapy: depression in the elderly--issues and advances in treatment. AB - Depression continues to be major cause of morbidity and mortality in the elderly. It is estimated that 1-5% of elderly persons who live in the community and 5-43% of nursing-home patients have major depression. Symptoms of depression in the elderly do not differ substantially from younger patients. Tricyclic antidepressants continue to be the drugs of choice in the elderly because of their long record of use with proven efficacy, known adverse effect profile and availability of less expensive generic formulations. The newer second-generation antidepressants, including serotonin reuptake inhibitors, appear to offer a major advantage of fewer serious adverse effects in the elderly. This review will highlight recent developments regarding the prevalence and treatment of depression in the elderly. PMID- 8227233 TI - A comparison of the beta 1-selectivity of conventional metoprolol and metoprolol CR during exercise in healthy volunteers. AB - This paper reports on a randomized double-blind crossover study to compare the effects of daily treatment with 100 mg conventional metoprolol (M100), 50 or 100 mg slow-release metoprolol CR (CR50, CR100) and placebo on the response to treadmill walking exercise in 12 healthy volunteers. Twenty minutes of exercise was undertaken at 1.5 h post-dose (Ex1) and again at 4 h (Ex2). During Ex1 all metoprolol preparations caused a reduction in the exercise-induced increase in heart rate when compared to placebo (P < 0.001). The exercise-induced increase in plasma glycerol, an indicator of lipolysis, was diminished by all metoprolol preparations but to a greater extent by M100 (P < 0.05). Rises in plasma ammonia were used to assess the net degradation of energy-rich adenine nucleotides by skeletal muscle during exercise. When compared to placebo, a significant elevation in plasma ammonia was seen only during exercise with M100 (P < 0.05). The perceived exertion at the end of exercise was also greatest on conventional metoprolol. The differences between the effects of conventional and CR formulations of metoprolol were less when exercise was repeated 4 h post dosing. We conclude that metoprolol CR may offer advantages over the conventional preparation, in subjects regularly engaging in exercise, by reducing the metabolic stress imposed upon skeletal muscle. PMID- 8227234 TI - Dosage programming of phenobarbital in neonatal seizures. AB - Owing to the changes occurring in the organism as a result of biological maturation, disposition kinetics of phenobarbital in newborns is significantly different to that observed in the paediatric and adult populations. Moreover, the disposition parameters change constantly during the first days of life. The data on the serum levels of phenobarbital in 17 newborns were analysed to quantify the changes in the elimination half-life of phenobarbital during the first weeks of life. The half-life of the drug was estimated to be (mean +/- SD) 114.2 +/- 43.0 h, 73.19 +/- 24.17 h and 41.23 +/- 13.95 h in patients 1-10, 11-30 and 31-70 days old, respectively. According to these values and assuming phenobarbital serum levels of 20 mg/l to be safe and effective in neonatal seizures, the initial dosing recommended is 2.9, 4.8 and 6.0 mg/kg/day in newborns 1-10, 11-30 and 31 70 days old, respectively. PMID- 8227235 TI - Relationship between slime production, antibiotic sensitivity and the phagetype of coagulase-negative staphylococci. AB - Three hundred and three strains of coagulase-negative staphylococci (CNS) were collected from the fingers of healthy donors (289) and from blood cultures (14). Twelve different species were identified (5 S. auricularis, 45 S. capitis, 15 S. cohnii, 86 S. epidermidis, 23 S. haemolyticus, 37 S. hominis, 1 S. lentus, 5 S. saprophyticus, 7 S. sciuri, 6 S. simulans, 54 S. xylosus and 19 S. warneri). Amongst these CNS strains, 151 were slime producers, 112 were phage-typable and 188, 133, 126 and 91 were, respectively, resistant to penicillin, teicoplanin, erythromycin and kanamycin. Slime-producing strains were resistant to at least seven antibiotics with a probability of 0.01 < P < 0.05. Non-slime-producing strains were sensitive to all the tested antibiotics with a probability of 0.001 < P < 0.01. There was no direct relationship between antibiotic sensitivity and phage type, although a non-typable strain was more often resistant to seven or more antibiotics than a typable one (0.05 < P < 0.1). PMID- 8227236 TI - Factors influencing pharmacists' preference for the legal classification of Rx-to OTC switched drug products. AB - The United States is one of the few developed countries that has only two legal classifications of drug products, prescription (or legend) and non-prescription. U.S. pharmacy associations, however, are lobbying for legislation that will classify drugs which are switched from prescription to non-prescription (Rx-to OTC) status as a third class of pharmacist-legend or pharmacy-only drug products. Pharmacists have indicated that their demand for a third class of drugs is to enable safer use of switched drug products by consumers through pharmacist supervision. Critics of the third class concept, however, have attributed the pharmacists' demand to a desire to gain economic control of the products. In order to assess empirically pharmacists' motivations, data collected from 389 randomly selected U.S. pharmacists, forming part of a larger study, were used to assess: (i) pharmacists' preferences for five selected conditions of sale for drug products switched from prescription to non-prescription status and (ii) the role of selected demographic and practice characteristics on pharmacists' preferences. Analyses of the data indicated that 'permanent pharmacist supervised sale (or a third class status)' and 'general unsupervised' sale were pharmacists' most and least preferred conditions of sale for switched products, respectively. Overall, pharmacists preferred conditions of sale that involved pharmacist supervision over those that did not. Pharmacists' preferences differed significantly when compared on the basis of their employment status. Owner or partner pharmacists indicated a greater preference for the pharmacist-supervised third class status for switched drug products and a lesser preference for unsupervised sale of switched drug products than staff, employee, director, or manager pharmacists. Pharmacists' preferences for the conditions of sale also differed by the nature of the professional association in which they were members. Pharmacists who were members of the American Society of Hospital Pharmacists (ASHP) indicated a greater preference for 'unsupervised sale after initial diagnosis and prescription from a physician', than pharmacists who were not members of the association. Pharmacists who were members of the National Association of Retail Druggists (NARD) viewed unsupervised sale of switched products with greater disfavour than the non-members of that association. Overall, it appears that some pharmacists may be governed by safety concerns and some by economic concerns in their demand for a third class or pharmacy-only status for Rx-to-OTC switched drug products. PMID- 8227237 TI - The development of oral liquid dosage forms of metronidazole. AB - The stability of eight samples of metronidazole in aqueous vehicles was studied using a modified HPLC assay method. The samples were prepared using either metronidazole powder (5 mg/ml), metronidazole hydrochloride injection powder (10 mg/ml), or metronidazole powdered tablets (10 mg/ml). The samples prepared using metronidazole hydrochloride powder were stable and clear for at least 133 days at 25 degrees C. Their pH values were between 1.8 and 2.0. The samples prepared from metronidazole powder had pH values close to neutral and were stable for at least 45 days at room temperature (25 degrees C). However, to keep the drug in solution, the concentration of metronidazole powder was reduced to 5 mg/ml. The powdered tablets did not yield good uniform suspensions. PMID- 8227238 TI - Effect of value-added utilities on prescription refill compliance and Medicaid health care expenditures--a study of patients with non-insulin-dependent diabetes mellitus. AB - An estimated 20 million Americans suffer from diabetes. Patients with non-insulin dependent diabetes mellitus (NIDDM) comprise approximately 90% of the diabetic population. An estimated 10-30% of patients with NIDDM withdraw from their prescribed regimen within 1 year of diagnosis, and of the remainder, nearly 20% administer insufficient medication to facilitate an adequate reduction in blood glucose. A randomized trial was undertaken to discern the effect of pharmacy based value-added utilities on prescription-refill compliance with sulfonylurea therapy and health service utilization. The subjects were 258 Medicaid beneficiaries from the state of South Carolina, previously untreated for NIDDM, prescribed 5 mg of the second-generation sulfonylurea glyburide twice daily, and monitored with regard to prescription-refill compliance and health service utilization for 1 year. Subjects provided informed consent and were randomly assigned to one of four experimental groups: (i) the control cohort received standard pharmaceutical care with each dispensing of glyburide; (ii) the second cohort received standard pharmaceutical care and was mailed a medication-refill reminder 10 days prior to each sequential refill date; (iii) the third cohort received standard pharmaceutical care and was provided unit-of-use packaging with each prescription-refill request; (iv) the fourth cohort received standard pharmaceutical care, mailed medication-refill reminders, and unit-of-use packaging. Analysis of variance (ANOVA) procedures revealed that patients receiving mailed prescription-refill reminders, unit-of-use packaging, or a combination of both interventions achieved a significant (P < or = 0.05) increase in the Medication Possession Ratio (MPR) for sulfonylurea therapy relative to controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227239 TI - Simultaneous measurement of chloramphenicol sodium succinate and chloramphenicol in presence of furosemide in plasma and urine. PMID- 8227240 TI - Myocarditis or acute myocardial infarction? PMID- 8227241 TI - While the getting's good. PMID- 8227242 TI - Management of varices in cirrhosis. PMID- 8227243 TI - The cellular basis of Fanconi syndrome. AB - In this syndrome, all solutes are prevented from crossing the apical membrane of the proximal renal tubule cell; hence, investigations have focused on generalized transport dysfunction, namely intracellular ATP activity. Experiments suggest that intracellular adenine nucleotide degradation secondary to phosphate depletion causes a decline in ATP. PMID- 8227244 TI - Night sweats redux. PMID- 8227245 TI - Night sweats redux. PMID- 8227246 TI - Treatment of rheumatoid arthritis. PMID- 8227247 TI - Chronic diarrhea and vomiting in an elderly woman. PMID- 8227248 TI - Thrombolysis and stroke management. AB - Up to 90% of patients (those with occlusive stroke) may be candidates for clot lysis. Fears that lysis might cause hemorrhage appear to be unwarranted. The most important clinical lesson learned to date is that the earlier the intervention, the better the outcome. Issues such as the most efficient diagnosis, most amenable sites, and IV versus intra-arterial administration are under study. PMID- 8227249 TI - Immune regulation: learning from leprosy. AB - The specific anergy seen in severe, disseminated leprosy appears to represent multifaceted actions of macrophages and suppressor T lymphocytes. Identification of cytokine mediation in these events helps to account for the immune system's "choice" of cellular versus humoral defenses--and points toward manipulation of cytokine switches for immunotherapy against infectious and autoimmune disease. PMID- 8227250 TI - Diagnosis and management of peripartum cardiomyopathy. PMID- 8227251 TI - Renal tubular transport disorders. PMID- 8227252 TI - Non-radioactive detection methods for nucleic acids separated by electrophoresis. AB - The different non-radioactive labelling and detection methods currently commercially available are compared and evaluated in this review. Minor factors such as electrophoresis and blotting techniques as well as choice of membrane and their impact on results are discussed. Two major labelling moieties, biotin and digoxigenin, and the various labelling methods are discussed in detail. A comparison of my own results and those from the literature favours application of the digoxigenin group as a routine label. Nevertheless, in several cases biotin will also lead to good results and may also serve as a second label. The most important factor within the non-radioactive systems is the detection of the targeted label. Colorimetric and chemiluminescent techniques are compared in terms of sensitivity, flexibility and applicability. Colorimetric detection can produce suitable results, but in most cases the major advantages of chemiluminescent techniques involving alkaline phosphatase and AMPPD or CSPD will make chemiluminescent detection the method of choice. A survey is given on applicability of the basic techniques to several important assay methods involving electrophoresis of nucleic acids. Finally, some examples of application of non-radioactive nucleic acid labelling and detection techniques in plant molecular biology and biomedicine are cited from the literature. PMID- 8227253 TI - Activity gel and activity blotting methods for detecting DNA-modifying (repair) enzymes. AB - Zymographical methods (activity gel, overlay gel, activity blot and activity blotting) for detecting DNA-modifying (repair) enzymes are reviewed. Emphasis is put on the novel activity blotting method in which DNA repair enzymes electrophoresed on a gel are blotted and detected on a damaged DNA-fixed nylon membrane. Its practical procedures, including a non-radioactive detection procedure, and representative results are also described. PMID- 8227254 TI - Reversed-phase high-performance liquid chromatography of purine compounds for investigation of biomedical problems: application to different tissues and body fluids. AB - An overview of high-performance liquid chromatographic separation techniques (reversed-phase and ion-pair reversed-phase) used in the analysis of purine ribonucleotides, ribonucleosides and nucleobases, including procedures for sample preparation, is given. Coverage of the separation techniques is extended to the measurement of specific radioactivities of these compounds in tracer kinetic experiments for metabolic flux rate analyses. This article is focused on the development and adaptation of reversed-phase separation techniques for nucleotides, nucleosides and bases that are used to examine a variety of biomedical problems. The investigation of purine nucleotide metabolic disorders or physiological transitions in the pathomechanisms of different diseases and syndromes or in cell maturation processes, respectively, requires the application of chromatographic separation to a multitude of tissues and body fluids. These samples vary greatly in concentrations of purine compounds with low molecular mass, from ca. 5 mM to ca. 0.5 microM. The advantages and disadvantages of different techniques are critically discussed. PMID- 8227255 TI - Checking of individuality by DNA profiling. AB - A review of methods of DNA analysis used in forensic medicine for identification, paternity testing, etc. is provided. Among other techniques, DNA fingerprinting using different probes and polymerase chain reaction-based techniques such as amplified sequence polymorphisms and minisatellite variant repeat mapping are thoroughly described and both theoretical and practical aspects are discussed. PMID- 8227256 TI - Quantitative analysis of enzymic digests of DNA using gel electrophoresis. AB - Quantitative electrophoresis of enzymic digests of DNA and its applications are reviewed. Factors affecting the overall analysis such as DNA length distribution, labels for visualizing DNA, techniques for quantitation, and electrophoresis itself are studied. Methods to analyze restriction fragments and lesions in DNA by gel electrophoresis are presented. PMID- 8227257 TI - Size-dependent chromatographic separation of nucleic acids. AB - Chromatographic procedures currently used for the size-dependent fractionation of nucleic acids are reviewed. First, an attempt is made to clarify the concept of "size" of nucleic acids and then various aspects of the chromatography of nucleic acids are considered. It is emphasized that consideration of the dynamic three dimensional structure of large polynucleotides in a rapidly flowing eluent is essential for both the better understanding of mechanism and the development of sophisticated procedures. Of the practical chromatographic techniques that are not based on true size fractionation, ion-exchange chromatography on non-porous column packings appears to be the most efficient. Other methods, such as hydrophobic interaction, are unlikely to become popular. As for truly size dependent modes, there are gel permeation and slalom chromatography. Although media with extremely large pores become available, the efficiency of gel permeation is still low as a practical separation procedure for large nucleic acid molecules. Its best use seems to be in the field of physicochemical research into nucleic acids in solution. The newly discovered slalom chromatography is based on a principle completely different from all other chromatographic modes. It enables the efficient separation of large double-stranded DNA fragments of 5 50 kilo base pairs by discriminating their length. It has proved not only to be useful as a tool for nucleic acid research but also to have great significance in other fields, e.g. the hydrodynamics of polymer solutions, the search for new chromatographic modes, etc. PMID- 8227258 TI - Field alternation gel electrophoresis--status quo. AB - Since the description of the original technique of field alternation gel electrophoresis (FAGE) about ten years ago there have been significant developments in the area. Between 1983 and early 1987 dramatic improvements in the technique and apparatus resulted in a 500- to 600-fold increase in the functional separation capacity of conventional agarose gel electrophoresis. Details of the improvements in technique and equipment was the subject of an earlier review [H. J. S. Dawkins, J. Chromatogr., 492 (1989) 615]. This review concentrates on the application of FAGE technology. The FAGE technique is no longer restricted to simply separating large DNA fragments. This method is presently being used for electrophoretic karyotyping, long-range genomic mapping, cloning of large DNA fragments into new vectors, the study of pathogenic chromosomal alterations and the structural analysis of chromosomes. The applications of FAGE in molecular biology and genetics is constantly expanding, with the full potential of this technique still to be realised. PMID- 8227259 TI - Application of high-performance liquid chromatography for recognition of covalent nucleic acid modification with anticancer drugs. AB - Covalent modification of DNA by antineoplastic agents represents a potent biochemical lesion which can play a major role in drug mechanism of action. The ability to measure levels of DNA covalent modifications in target cells in vivo may, therefore, be seen as the ultimate form of therapeutic drug monitoring. Additionally, elucidation of the structure of critical DNA adducts and definition of their role in tumour cell cytotoxicity will provide more selective targets for rational drug design of new cancer chemotherapeutic agents. High-performance liquid chromatography has contributed significantly to all these areas. In vivo levels of nucleic acid covalent modifications are in the range of 1 in 10(5) 10(8) nucleotides precluding the use of conventional high-performance liquid chromatographic detection methods. Several classes of natural product anticancer drugs have been shown to bond covalently to nucleic acids under optimal laboratory conditions. These have proved more accessible to high-performance liquid chromatographic analysis because of their lipophilicity and strong UV chromophores. However, the majority of experimental evidence to date suggests that with the exception of mitomycin C and morpholino-anthracyclines these compounds do not exert their primary mechanism of action through nucleic acid covalent modification. DNA adducts of alkylating and platinating agents are more difficult to detect by high-performance liquid chromatography and can be chemically unstable. These compounds interact with DNA on the basis of chemical kinetics. Thus, the principle sites of attachment tend to be with the most nucleophilic base (guanine) at its most reactive centre (N-7 position). Limited in vivo high-performance liquid chromatographic studies with all classes of anticancer drugs indicate a much more complex pattern of adductation than would have been anticipated from in vitro studies alone. Some of these differences are probably due to methodological artefacts but these studies stress the need for sensitive detection methods and reliable sample preparation (nucleic acid extraction and digestion techniques) when attempting to determine nucleic acid covalent modifications by anticancer drugs. PMID- 8227260 TI - Electrophoretic and chromatographic separation methods used to reveal interstrand crosslinking of nucleic acids. AB - Several electrophoretic and chromatographic techniques, many of which have only been developed recently, provide sensitive methods for the detection and separation of DNA containing interstrand crosslinks such as those produced by many cancer chemotherapeutic drugs and photoactive psoralen derivatives. Most of the methods rely on the fact that the presence of such crosslinks prevent the complete denaturation of the two complimentary DNA strands by heat or alkali. A simple and highly sensitive neutral agarose gel electrophoresis method is particularly applicable to detailed time-course experiments of both total crosslink formation, and the "second-arm" of the crosslink reaction. This method separates denatured single-stranded from double-stranded DNA which has reannealed as a result of an interstrand crosslink. Polyacrylamide gel-based assays using denaturing gels are more suited to the separation of smaller crosslinked DNA fragments and, in particular, small oligonucleotides on high-percentage gels. In addition, they provide methods for the determination of the exact base position and sequence selectivity of crosslink formation. Sephadex chromatography and high performance liquid chromatography can separate small crosslinked oligonucleotides from non-crosslinked duplexes, and the hydroxyapatite column chromatographic separation of single- and double-stranded cellular DNA can be used to quantitate the level of interstrand crosslinking present in the bulk of the genome. Finally, the analysis of damage by crosslinking agents, and its repair, at the level of specific genes can be achieved by hybridization with specific probes following membrane transfer from neutral agarose gels used to fractionate restricted and fully denatured genomic DNA from drug-treated cells. PMID- 8227261 TI - Methods used for analyses of "environmentally" damaged nucleic acids. AB - In this review, we present various techniques, currently applied in many laboratories, which are useful in the detection of "environmentally"-induced damage to DNA. These techniques include: (a) chromatographic methods, which allow determination of chemical changes within DNA, be they formation of adducts with or oxidation of bases in DNA; (b) electrophoretic methods, which facilitate finding the site(s) in DNA where that chemical modification occurred; and (c) immunological assays, which help to detect DNA damage using externally produced antibodies that recognize the specific chemical changes in DNA or its fragments, as well as by detection of autoantibodies that develop in response to environmental exposures of animals and humans. PMID- 8227262 TI - Reversed-phase and ion-pair separations of nucleotides, nucleosides and nucleobases: analysis of biological samples in health and disease. AB - Methods for the assay of nucleotides, nucleosides and nucleobases in biological samples in health and disease are reviewed, with emphasis on reversed-phase and ion-pair reversed-phase techniques for their determination. Modes of extraction from biological samples are discussed with respect of the determination of in vivo concentrations. Advantages and limitations of ion-pair reversed-phase chromatography are discussed with examples from biochemistry and clinical chemistry. The capacity of the high-performance capillary electrophoresis is compared with that of ion-pair reversed-phase chromatography. PMID- 8227263 TI - Affinity chromatography with nucleic acid polymers. AB - Column chromatography utilizing polynucleotides immobilized on solid support is reviewed. This form of affinity chromatography is used for the isolation of polynucleotides and polynucleotide binding proteins, and to a lesser extent for analysis. Several specific applications within these categories have been widely used in the biomedical sciences. Poly(A) mRNA is routinely isolated using oligo(dT) or oligo(dU) supports. Many DNA binding proteins, including transcription factors, restriction endonucleases, and proteins involved in DNA repair, replication, recombination, and transposition have been purified using DNA affinity chromatography. Recently, DNA supports suitable for use in high performance liquid chromatography have been described and utilized. The current usage of DNA affinity chromatography is reviewed and potential future uses for this technology are speculated upon. PMID- 8227264 TI - Prediction of the behaviour of oligonucleotides in high-performance liquid chromatography and capillary electrophoresis. AB - The present state of studies on prediction of the chromatographic and electrophoretic behaviour of oligonucleotides is reviewed; particular emphasis is given to high-performance liquid chromatographic and capillary electrophoretic separations. Attention is paid to fundamental theory for the prediction of retention and migration times, and bandwidths. The article also deals with the applicability of the theory to the computer-assisted prediction and the computer simulation for these two types of separation of oligonucleotides. Optimization of separation conditions using the computer simulation system is briefly described. PMID- 8227265 TI - Isotachophoresis of nucleic acid constituents. AB - Determination methods for purine and pyrimidine bases, nucleosides, nucleotides and related compounds using analytical capillary isotachophoresis are reviewed. First, the isotachophoretic characterization of these compounds, as well as methods for sample preparation prior to analysis, and the different ways of detecting unknown substances in complex biological systems are described. Then applications of isotachophoretic analysis in medical diagnosis and biomedical research are reviewed. In particular, the analysis of purines, pyrimidines, nucleosides and related compounds in blood and serum for the diagnosis and treatment of inherited diseases and cancer is described. Selected applications of nucleotide analysis in biomedical research using different tissue extracts are also reviewed, and some examples of nucleotide-dependent enzymic reactions, which were performed by means of analytical isotachophoresis, are presented. PMID- 8227266 TI - Comparison of properties of agarose for electrophoresis of DNA. AB - Agarose as a medium for separation of DNA was first introduced in 1962 and since the early 1970s agarose submarine gel electrophoresis has been synonymous with separations of DNA molecules larger than 1 kilobase pair (kb). The large pore size, low electroendosmosis and strength of the matrix have advantages over other media such as polyacrylamide for many applications. The variety of grades of agarose, developed by chemical manipulation of the substitutions on the agarose polymer, provides a range of matrices for separation of DNA molecules from a few base pairs (bp) to over 5 megabase pairs (Mb) in length. The introduction of low melting-temperature agarose has revolutionised the extraction and manipulation of chromosome-sized molecules. On the other hand, the demand for analysis of very small quantities of DNA will most likely lead to the increasing importance of capillary electrophoresis. Many theories have been propounded to explain the electrophoretic migration of DNA in agarose. The most popular of these has been reptation theory but none can account for all of the reported anomalies in migration. However, anomalous migration has been exploited to study DNA structure, topology and catenation. An example of the use of two-dimensional electrophoresis to demonstrate the complexity of DNA migration through agarose is given. Generally, for molecules smaller than 50 kb, electrophoretic separation is a function of length. By alternately electrophoresing DNA in two different directions, molecules as large as 5.7 Mb have been effectively separated, although with such large molecules DNA structure as well as size may determine migration. In the case of separations of chromosomes from the intestinal protozoan, Giardia duodenalis, for example, a discrepancy of 1 Mb in the size of one chromosome, with an apparent size of 0.7-2.0 Mb, depended on the boundary conditions of separation. Major challenges for the molecular biologist are separation of larger chromosomal sized molecules, greater number of samples and smaller formats. Towards this challenge computer-aided technology is a key component in the control of electrophoresis parameters and analysis. PMID- 8227267 TI - Development of a high-performance liquid chromatographic-mass spectrometric technique, with an ionspray interface, for the determination of platelet activating factor (PAF) and lyso-PAF in biological samples. AB - An HPLC-mass spectrometric technique with an ionspray interface was developed for the determination of platelet-activating factor (PAF) and PAF-related compounds in biological samples. HPLC separations were performed using a reversed-phase column. The mass spectra showed intense [M + H]+ ions. Collision-induced dissociation of protonated molecular ions gave characteristic daughter ions corresponding to the phosphorylcholine group. By selective-ion monitoring, a detection limit of 0.3 ng was obtained for all molecules; by multiple reaction monitoring, the same sensitivity was achieved for PAF whereas for lyso-PAF the limit was 3 ng. Finally, PAF was comparatively determined by bioassay and HPLC-MS after extraction from the cell pellets and the supernatants of human polymorphonuclear neutrophils unstimulated or stimulated with opsonized zymosan. The good correlation observed between these techniques indicated the reliability of HPLC-MS for biochemical studies on PAF and PAF-related molecules. PMID- 8227268 TI - Characterization of the chemical structure of sulphated glycosaminoglycans after enzymatic digestion. Application of liquid chromatography-mass spectrometry with an atmospheric pressure interface. AB - Pneumatically assisted electrospray was demonstrated to be a powerful ionization source for the analysis of oligosaccharides. A mass spectrometer was interfaced to an HPLC system, using this interface, to determine oligosaccharides from the enzymatic digestion of heparin separated on a reversed-phase column. To set up the technique, and particularly to clarify the ionization process, purified disaccharides, from enzymatic digestion of chondroitin sulphates, were measured. The use of a suitable counter ion in the mobile phase, tetrapropylammonium (TPA), to optimize the HPLC separation, gave, with sulphated di- and oligosaccharides, adducts [M + nTPA - (n + m)H]m-, which were unexpectedly stable to fragmentation; molecular ions [M - (n + 1)H]n-, in the presence of the counter ion, were observed only with desulphated or monosulphated disaccharides. The stability of the adducts and the use of a deuterated ion-pair reagent permitted an exact evaluation of the molecular masses of disaccharides and oligosaccharides of unknown structure. Spectra obtained in the absence of the counter ion contained singly or multiply charged molecular ions and fragmentation ions mainly from loss of the sulphate groups; under these ionization conditions the exact mass determination and interpretation of the spectra were difficult. After removal of the counter ion, tandem mass spectra could be obtained with some interesting data for the characterization of these molecules. Complete spectral analyses were performed with amounts of samples of 50 micrograms but, using microbore columns, one twentieth of this amount may give good spectra. PMID- 8227269 TI - Rapid analysis of enzymatic digests of a bacterial protease of the subtilisin type and a "bio-engineered" variant by high-performance liquid chromatography frit fast atom bombardment mass spectrometry. AB - Amino acid sequencing of a subtilisin-type bacterial protease and a bio engineered variant was carried out by investigating various enzymatic digests using HPLC-frit fast atom bombardment MS methods. The fast atom bombardment mass spectral data allowed rapid identification of the enzymatically generated peptides and differentiation between both proteins. The feasibility of determining the positions and nature of mutations in the amino acid sequence depends mainly on the size of the peptides containing the modifications. PMID- 8227270 TI - Pseudo-electrochromatography--negative-ion electrospray mass spectrometry of aromatic glucuronides and food colours. AB - Pseudo-electrochromatography is a combination of liquid chromatography and an electromigration technique, especially directed at the separation of ionic compounds prior to mass spectrometric detection with a mobile phase composition compatible with mass spectrometry. The application of pseudo electrochromatography to the separation of food colours and aromatic glucuronides is described. An example of selectivity tuning by applying voltages of differing polarity during the chromatographic run is given. The coupling of pseudo electrochromatography with electrospray mass spectrometry is demonstrated. Differences in the effects of the axial potential over the column between silica based and polymeric packing materials are discussed. PMID- 8227271 TI - Simple high-performance liquid chromatographic method for assessing the deterioration of atropine-oxime mixtures employed as antidotes in the treatment of nerve agent poisoning. AB - A set of reversed-phase HPLC conditions for determining the degradation of atropine and the oxime (pralidoxime, obidoxime, or HI-6) in autoinjectors designed for use against nerve agent poisoning is described. The assay conditions for atropine do not require its prior separation from the large molar excess of oxime since both the atropine and tropic acid peaks elute well clear of the oxime and its degradation products and the phenolic preservatives. Further dilution of the sample and simple changes to the mobile phase then provide conditions for the oxime and its major degradation products to be quantitated. PMID- 8227272 TI - Characterization of metallothionein isoforms. Comparison of capillary zone electrophoresis with reversed-phase high-performance liquid chromatography. AB - The purpose of this study was to compare and contrast the separation of metallothionein (MT) isoforms by reversed-phase high-performance liquid chromatography (RP-HPLC) with capillary zone electrophoresis (CZE). RP-HPLC was performed on a Vydac C8 column eluted with a linear acetonitrile gradient. CZE was performed in a 57 cm x 75 microns I.D. fused-silica tube at an operating voltage of 30 kV. Phosphate buffer (10 mM) at pH 2.5, 7.0 and 11.0 was used for both separations. CZE at pH 2.5 resolved three distinct peaks of rabbit liver MT which were incompletely resolved at pH 7.0 or 11.0. RP-HPLC at pH 2.5 gave two peaks and the resolution was not as good as with CZE at the same pH. At pH 7.0 or 11.0, RP-HPLC of rabbit liver MT gave a single predominant peak of unresolved MT 1 and MT-2. Purified rabbit liver MT-1 and MT-2 were used to verify the identity of these peaks. In contrast, MT from horse kidney demonstrated three predominant peaks which were best resolved by CZE at pH 11.0, whereas RP-HPLC resolved only two peaks at pH 11.0 and 7.0. At pH 2.5, RP-HPLC of horse kidney MT gave three peaks, though two of the peaks were incompletely separated. We conclude that pH has a considerable impact on the resolution of MT isoforms by CZE and RP-HPLC and that it is possible to exploit changes in pH to optimize the separation of isoforms for a particular species of MT. When samples of human and sheep liver MT 1, both of which exhibit microheterogeneity, were subjected to CZE, a single predominant peak was observed at each pH value. RP-HPLC of human liver MT-1 at pH 2.5 yielded two peaks that were incompletely resolved. Purified chick liver MT and rat liver MT-1 and MT-2 gave a single predominant peak at all pH values on CZE. In contrast, pig liver MT-1 and MT-2 each exhibited multiple peaks when subjected to CZE, the number of which depended on the pH used to separate the MT. In conclusion, CZE, with its orthogonal selectivity, and RP-HPLC make an excellent combination for the separation and characterization of MT isoforms. Because CZE is rapid (run times typically < 10 min) and requires little sample (< 100 nl), MT samples can readily be analyzed by CZE in conjunction with RP-HPLC or other techniques in order to maximize the information obtained about the individual isoforms. PMID- 8227273 TI - A novel, sensitive radioimmunoprecipitation assay for the detection of antibodies to human immunodeficiency virus-type 2. AB - A radioimmunoprecipitation assay that provides an alternative to the Western blot assay was developed for characterizing antibodies against human immunodeficiency virus type-2 (HIV-2). The assay is based on radioiodination of antigen using Bolton-Hunter reagent. The antigen consists of a soluble preparation of the NIH-Z (HIV-2) strain of 1000X purified virus spiked with purified recombinant HIV-2 gp105. Radiolabeled proteins were immunoprecipitated by immune human sera, even at the early stages of seroconversion. This new assay provides a simple method for characterizing and titrating antibodies against HIV-2. The method is more sensitive, and is more efficient than Western blotting. The labeled viral proteins are well suited for biochemical studies. PMID- 8227274 TI - Sweet potato feathery mottle potyvirus (C1 isolate) virion and RNA purification. AB - A procedure for the purification of a Peruvian isolate (C1) of sweet potato feathery mottle potyvirus (SPFMV) and infective RNA has been developed. The use of Hepes [N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid] buffer containing urea and sodium EDTA as a base for tissue extraction and virus suspension enabled good yields of virus (35-50 mg/100 g) to be obtained from Nicotiana benthamiana L. Domin. A short RNA isolation procedure yielded infectious RNA, from which ds cDNA of nearly genome size could be obtained. Sweet potato feathery mottle potyvirus, Purification, RNA isolation, cDNA synthesis. PMID- 8227275 TI - Detection of human parvovirus B19-specific IgM and IgG antibodies using a recombinant viral VP1 antigen expressed in insect cells and estimation of time of infection by testing for antibody avidity. AB - Sera from patients with symptoms of recent human parvovirus B19 (B19) infection were tested for B19-specific IgM in an immunofluorescence assay (IFA) using insect cells expressing B19 recombinant VP1 coat protein as an antigen. A highly significant correlation (P < 0.001) was found between titres obtained in the IgM IFA and the units obtained in an IgM antibody-capture RIA using plasma derived native B19 antigen. An IgG IFA using the recombinant antigen was performed on 57 sera and the antibody avidity determined. There was a highly significant correlation (P < 0.001) between the relative amounts of low avidity B19-specific IgG antibodies and time after onset of illness. This finding allows the detection of IgG to be used for diagnosing acute infection. PMID- 8227276 TI - Detection of bovine herpesvirus-1 in bovine semen by a nested PCR assay. AB - A nested PCR assay targeting a portion of the glycoprotein IV gene has been developed for the detection of Bovine Herpesvirus-1 (BHV-1). Rapid and sensitive detection of the PCR products was achieved using a nonisotopic reverse dot-blot format with a visible color readout. Cross-reactivity of this PCR assay was not observed with the closely related BHV-3. The sensitivity of this assay when tested on a supernatant from a BHV-1 cell culture was approximately 4.5 TCID50 (50% tissue culture infectious dose). A procedure using the chelating resin Chelex 100 was used to prepare viral DNA from artificially inoculated samples of extended and raw semen for use in the PCR assay. In combination with nested PCR and reverse dot blot, this method allowed the detection of 5 x 10(3) TCID per 0.5 ml of semen, which is comparable to the detection in the Cornell Semen Test. The whole procedure can be completed in approximately 8 h. This assay has therefore the potential of replacing the currently available yet time consuming and costly detection methods for BHV-1 in bovine semen. PMID- 8227277 TI - Direct sequence determination of the influenza B HA-1 gene after PCR amplification of clinical specimens from an infected volunteer. AB - When clinical isolates of influenza A and B viruses are propagated in embryonated hens' eggs or tissue culture cells, different selective pressures in vitro result in specific amino acid substitutions in the haemagglutinin (HA) gene. A proportion of such viruses which lose a potential glycosylation site near the receptor binding region of the HA at amino acid positions 196-198 appear to have reduced virulence. Direct polymerase chain reaction (PCR) amplifications of cDNA and subsequent nucleotide sequence analysis of part of the HA-1 gene of the original infecting influenza B strain and the nasal wash material from an infected volunteer were performed. The nucleotide sequences of the viral HA-1 from the nasopharynx of the infected volunteer were the same as that of the original infecting strain. Antigenic analysis of both the original infecting virus and the viruses isolated from sequential samples collected from the volunteer, all of which were cultivated on Madin-Darby Canine Kidney (MDCK) cells and in embryonated hens' eggs, revealed variation in the HA of viruses only after egg adaptation. In particular, we describe the use of direct nucleotide sequencing techniques without the use of cloning strategies in order to determine the sequence of the HA-1 gene after direct PCR amplification of clinical nasal wash material. PMID- 8227278 TI - Quantification of human cytomegalovirus DNA in peripheral blood polymorphonuclear leukocytes of immunocompromised patients by the polymerase chain reaction. AB - Human cytomegalovirus (HCMV) DNA amplification by the polymerase chain reaction (PCR) was utilized previously for successful monitoring of HCMV infections in immunocompromised patients. However, analysis of an extended series of clinical samples revealed the relatively frequent presence of PCR inhibitors. Hence, the need for availability of an internal control of the reaction allowing identification of false negative results. Similarly, an internal standard appeared necessary for quantification of viral DNA in clinical samples. For this purpose, we constructed a recombinant DNA molecule which could be amplified by the same set of primers used for HCMV DNA amplification. Coamplification of the recombinant DNA molecule and clinical samples proved to be a simple and reliable method for verifying sample competence for amplification. In addition, coamplification of serial known amounts of the same molecule, used as internal standard, and test sample, allowed quantification of viral DNA in polymorphonuclear leukocyte samples. Quantitative monitoring of HCMV infection and antiviral treatment may provide critical indications as to whether and when to initiate or discontinue antiviral treatment in immunocompromised patients with systemic HCMV infections. PMID- 8227279 TI - Detection of HIV-1 by digoxigenin-labelled PCR and microtitre plate solution hybridisation assay and prevention of PCR carry-over by uracil-N-glycosylase. AB - An extremely sensitive and convenient microtiter plate solution hybridisation assay for the detection of HIV-1 PCR products was developed. The PCR product is labelled by direct incorporation of digoxigenin-dUTP and after denaturation is captured by a microtitre plate coated with a streptavidin-linked biotinylated probe. The PCR/probe hybrids are reacted with an alkaline phosphate conjugated anti-digoxigenin antibody and detected using an alkaline phosphatase enzyme amplification system. The use of uracil-N-glycosylase and dUTP instead of dTTP in the PCR is used to effectively control carry-over from previous PCR products. The assay can detect single HIV-1 DNA molecules in a background DNA of 0.75 microgram. PMID- 8227280 TI - Simple microwave and thermal cycler boiling methods for preparation of cervicovaginal lavage cell samples prior to PCR for human papillomavirus detection. AB - Sample preparation is an important step in the detection of viral DNA by the polymerase chain reaction (PCR) technique. The method used should achieve release of cellular DNA with the minimum of manipulation steps so as to reduce the possibility of contamination. The present report demonstrates that either microwaving or 20 min of boiling in the heating block of a thermal cycler lead to satisfactory results in the detection by PCR of human papillomavirus in cervicovaginal epithelial cell specimens obtained by lavage. Since each of these methods uses only one step the possibility of contamination is greatly reduced compared to the widely used proteins K/nonionic detergent extraction procedure. PMID- 8227281 TI - Detection of human astrovirus serotype 1 by the polymerase chain reaction. AB - Astroviruses are small, plus-strand RNA viruses associated with diarrhoea, mostly in children. The diagnostic method commonly used is electron microscopy. We have designed a nested polymerase chain reaction (PCR) based on the recently reported nucleotide sequence of the 3' end of the genome of a human astrovirus serotype 1, the most common form. The PCR was positive for the ten serotype 1 samples tested, while being negative for all other viruses tested, including astrovirus type 2, 3, 4 and 5, calicivirus, rotavirus and picornaviruses. Fecal extracts from patients with diarrhoea were analysed directly or after isolation of RNA, the former method being at least as sensitive. Titration of fecal extracts by PCR indicated the presence of up to 10(11) viral particles per ml in feces. PMID- 8227282 TI - Horseradish peroxidase-labelled probes and enhanced chemiluminescence to detect baculoviruses in gypsy moth and eastern spruce budworm larvae. AB - Horseradish peroxidase-labelled whole genomic DNA probes and enhanced chemiluminescence procedures were utilized to detect baculovirus in insect macerates blotted on nylon membranes. Detection levels were similar to those found using 32P-labelled probes; 5 x 10(3) occlusion bodies (OBs), 2 x 10(3) OBs and 4 x 10(4) OBs of Lymantria dispar (L.) nuclear polyhedrosis virus (LdNPV), Choristoneura fumiferana (Clem.) NPV (CfNPV) and C. fumiferana granulosis virus (CfGV) respectively using 10 ng/ml LdNPV DNA probe and 20 ng/ml CfNPV and CfGV probe concentrations. Quantities of purified viral DNA detected were 0.56 ng LdNPV, 0.20 ng CfNPV and 0.10 ng CfGV at similar probe concentrations. Cross reactions were observed between LdNPV DNA probes and CfNPV. Multiple probing of membranes blotted with insect macerates was capable of diagnosing the presence of NPV and GV on membranes. This procedure appears to be useful in the diagnosis of large numbers of insects for several baculoviruses. PMID- 8227283 TI - Rapid identification and quantitation of cells infected by recombinant herpesvirus (pseudorabies virus) using a fluorescence-based beta-galactosidase assay and flow cytometry. AB - We recently described construction and use of a beta-galactosidase expression cassette in isolating recombinant pseudorabies virus (PrV) mutants (Mettenleiter and Rauh, 1990). We report here the identification and exact quantitation of cells infected by these mutants using an assay based on the reaction of intracellular beta-galactosidase expressed during infection by the recombinant viruses with the fluorogenic substrate fluorescein di-beta-D-galactopyranoside (FDG) followed by detection of positive cells in flow cytometry (FACS-Gal assay; Nolan et al., 1988). The detection method is fast, sensitive, and reliable, and yields quantitative results on single cell basis. PMID- 8227284 TI - The biology and pathology of Fc receptors. PMID- 8227285 TI - Surface expression of secretory component and HLA class II DR antigen on glandular epithelial cells from human endometrium and two endometrial adenocarcinoma cell lines. AB - The expression of secretory component (SC) by human glandular endometrial cells cultured in vitro was significantly increased by estradiol in the medium. Interferon-gamma and interleukin-4 stimulated the expression of SC only in the presence of estrogen. Tumor necrosis factor-alpha plus estrogen also caused a significant increase in the number of cells expressing SC. HLA class II antigen DR was detected on few glandular epithelial cells of human endometrium cultured in control medium, whereas interferon-gamma and interleukin-4, but not tumor necrosis factor-alpha, caused significant increases in the expression of DR. Estrogen in the culture medium did not significantly affect DR expression. The human endometrial adenocarcinoma cell lines, HEC and RL-95, expressed SC in approximately 50 and 20% of the cells. Also, approximately 20% of the RL-95 cells stained for DR antigen. Interferon-gamma did not influence the degree of expression of either surface marker of the two cell lines. Cells of both lines bound polymeric IgA and IgM but showed little to no binding of monomeric IgA, IgG, or an IgM previously shown not to bind SC. PMID- 8227286 TI - Long-term use of IgA-depleted intravenous immunoglobulin in immunodeficient subjects with anti-IgA antibodies. AB - The use of intravenous immunoglobulin is standard practice for antibody replacement in the humoral immunodeficiency diseases. Most infusions proceed uneventfully, but a proportion of infusions (5-8%) produces some degree of an infusion reaction. While the cause of most of these infusion reactions is unknown, an established, but rare cause of reactions is IgA antibodies in the serum of the patient, which apparently forms an immune complex with the traces of IgA in the infused immunoglobulin. This article describes our studies of five immunodeficient patients who had high-titered anti-IgA antibodies and a history of severe infusion reactions to intravenous immunoglobulin products not depleted of IgA (IgA content, 270-720 micrograms/ml). Over a 6-year period we gave these patients IgA-depleted intravenous immunoglobulin for a total of 170 infusions. These infusions were generally well tolerated; however, mild to moderate infusion reactions did occur in 9 of the 170 infusions (5.3%). These reactions were not related to the IgA content of the immunoglobulin solutions used--ascertained to vary between 0.4 and 2.9 micrograms/ml of IgA. Levels of plasma C3a and C4a increased after immunoglobulin infusions but the appearance of these components was not accompanied by any infusion reaction. We conclude that the long-term infusions of IgA-depleted intravenous immunoglobulin, within the range of IgA concentrations investigated, into patients with even very high-titered antibodies to IgA, is a safe practice. PMID- 8227287 TI - Abnormal craniofacial morphology and cartilage structure in transgenic mice harboring a Gly --> Cys mutation in the cartilage-specific type II collagen gene. AB - Gross morphology and histology of the craniofacial complex was studied in the offspring of two transgenic founder mice, Gly85-1 and Gly85-3, carrying several copies of a mouse type II collagen transgene that causes a single amino acid substitution (Gly-->Cys) at position 85 of the triple helix. The newborn transgenic mice had a short snout and mandible, a protruding tongue, a cleft of the secondary palate with the tongue situated between the shelves, and a doming cranial vault. Radiological examination revealed that the cranial base of the transgenic mice was shorter and its posterior part downward bent; in addition both the palate and the cribriform plate were less extended in relation to the cranial base than in the controls. Histologically the midline cartilaginous structures were composed of densely packed enlarged chondrocytes in a reduced extracellular matrix containing abnormally thick collagen fibrils. With the exception of the zone of hypertrophic chondrocytes the matrix also showed a loss of glycosaminoglycans. The cellular architecture of the basicranial synchondroses was disorganized, and the nasocerebrally oriented collagen fibrils formed unevenly distributed aggregates. The craniofacial morphology described here for the Gly85 mice shares features typical for other mouse mutations causing short limbed dwarfism. The observations indicate that defective cartilage production causes disproportionate craniofacial growth. Transgenic mice with specific mutations in cartilage-specific genes should therefore be useful for elucidating the complex mechanisms involved in determining the craniofacial growth. PMID- 8227288 TI - A morphometric analysis of human embryonic craniofacial growth in the median plane during primary palate formation. AB - As the human primary palate develops between embryonic stages 15 and 18, the facial prominences are part of a rapidly growing craniofacial complex that undergoes extensive morphogenetic change. The purpose of this study was to analyze growth in the medial plane in order to identify regional changes that occur during changes in craniofacial morphology. Photographs of midsagittal sections of 35 human embryos of stages 15 to 19 from the Carnegie Embryology Collection were enlarged, and landmarks were digitized for angular and linear measurements and for finite element modeling (FEM) analysis. The results showed magnitudes and directions of growth required to change average stage 15 morphology to later stages. As the facial and cranial components increased in size, shape change was most pronounced in the posterior cranial and orofacial regions. Increases in cranial linear dimensions were significantly larger than those in the cranial base regions. Between stages 15 and 18, the posterior cranial angle decreased, the forebrain and midbrain rotated superiorly toward the hindbrain, the orofacial angle increased, and the face grew above the thorax. The results suggest that morphogenetic growth changes in the cranial regions are closely associated with facial regions during primary palate formation. PMID- 8227289 TI - Growth and morphogenesis of the human embryonic midface during primary palate formation analyzed in frontal sections. AB - Although it is recognized that morphology of the craniofacial complex changes during primary palate formation, little information is available about the sites and amounts of growth in different regions. The purpose of this study was to analyze growth patterns of human embryonic heads in frontal sections and to identify regional growth associated with changes in craniofacial morphology and formation of the primary palate. Photographs of frontal sections of 31 human embryos of stages 16 to 19 from the Carnegie Embryology Collection were selected at seven different planes through each head, enlarged and traced, and landmarks were located for computerized morphometric measurements and for finite element modeling (FEM) analyses. Anatomical form change from the initial geometry of the average early stage 16 group to that of later stages was shown by numerical values and graphic displays of regional growth changes during midfacial morphogenesis. Between stages 16 and 19, extensive changes in midfacial morphology occurred as the frontonasal prominence elongated vertically (height increased by seven times) and narrowed to approximately half the width, with more pronounced changes in the anterior nasal region. The brain and the face became vertically separated, and the facial prominences became more frontally positioned relative to the brain. Changes in depth of the face were very limited during this period. The area of the maxillary region increased extensively, particularly in the anterior region, as the maxillary prominences grew forward, lateral to the nasal cavity, to contribute to the primary palate. The lateral nasal prominences increased in size with a predominantly horizontal growth pattern. These patterns of vertical growth of the midfacial tissues, narrowing of the nasal pits and frontonasal prominence, forward growth of the maxilla, and relative separation of the brain and face were identified as predominant features of embryonic craniofacial growth during primary palate development. PMID- 8227290 TI - Development of the basilar part of the occipital bone in normal human fetuses. AB - The present study aims to investigate the normal maturity and growth of the basilar part of the occipital bone evaluated radiographically and histochemically during the first half of prenatal life. The material consisted of cranial bases from 49 normal human fetuses and, in addition, three separate occipital bones deviscerated from normal fetuses. The normal development of the basilar part of the occipital bone was divided into five maturity stages (OB-I to OB-V). Microscopic investigation confirmed the macroscopic findings described. For each stage the ventro-dorsal dimension of the occipital bone component was measured (LEN). The stages of (OB-I to OB-V) were related to gestational age (AGE), crown rump length (CRL), LEN, and to the length of the humeral bone (HL). Close accordance was registered between OB stages and length values of the basilar part of the occipital bone, LEN, and length of the humeral bone, HL. A highly significant positive correlation was found between LEN and the parameters AGE, CRL, and HL values (P < 0.001). Standards for normal development of the basilar part of the occipital bone are considered important for evaluation of the pathological prenatal development of the cranial base. PMID- 8227291 TI - Computer reconstructions of human embryonic craniofacial morphology showing changes in relations between the face and brain during primary palate formation. AB - During early development of the human facial region, the craniofacial complex undergoes extensive morphogenetic change. The purpose of this study was to generate computer reconstructions of serially-sectioned embryos in order to illustrate major changes in spatial relations that occur between cranial and facial tissues. Five human embryos of stages 16 to 18 from the Carnegie Embryology Collection, which were used in the frontal morphometric study, were selected to illustrate phases of morphogenesis. Serial photographs of frontal sections were enlarged, traced, and digitized for computer reconstruction. The craniofacial complex was divided into components that were similar to those analyzed in a morphometric study [Diewert VM, Lozanoff S: J Craniofac Genet Dev Biol 1993: 13:162-183] to facilitate visualization of changes in regional components. The major changes observed included changes in form of the brain and its ventral contour above the face; enlargement, elongation, and more ventral positioning of the facial prominences relative to the brain; and forward positioning of the eyes and facial tissues relative to cranial components. As the brain became separated from the face at advancing stages of development, the frontonasal prominences narrowed and became vertically elongated, the lateral nasal prominences enlarged and became medially positioned, and the maxillary prominences enlarged and extended anteriorly. Changes in cranial flexion and lifting up and back of the forebrain [Diewert VM, Lozanoff S: J Craniofac Genet Dev Biol 1993:13:184-192.] appear to contribute to withdrawal of the brain from between the facial prominences during early midfacial development before cartilaginous tissues of the chondrocranium develop. PMID- 8227292 TI - Genetic and environmental effects on preimplantation development of mouse embryo in vitro. AB - To examine the degree of the relative contribution of genetic and environmental effects to preimplantation development of mouse embryo in vitro, two-cell stage embryos collected from oviducts of five strains of inbred mice (DDD/Qdj, C3H/Qdj, C57BL/6J Sea, DBA/1J Sea, and BALB/C Sea) were cultivated up to the stage of expanded blastocyst for 72 hr. Both fertility and survival rates were calculated in each recipient and also in each strain. A Student-Newman-Keuls test for both rates showed a significant inter-strain difference and an analysis of variance done to estimate the relative importance of genetic and environmental effects on the preimplantation development of embryo showed a low degree of genetic determination for both rates. It was thus possible to conclude that the environmental effect played a more important role on preimplantation development of mouse embryo in vitro. PMID- 8227293 TI - Intrauterine effect of dam on prenatal development of craniofacial complex of mouse embryo. AB - Embryo transfer effect and intrauterine effect of the dam on prenatal development of the craniofacial complex of mice embryos were investigated with the use of embryo transfer and cephalostat. DDD strain embryos were transferred to the three strains of recipients (DDD, C57BL, and DBA). The cephalometric observation of newborn offspring developed from transferred embryos was performed just after parturition. Dorso-ventral craniofacial size of newborn offspring was calculated using values of X- and Y-coordinates on a dorsoventral cephalogram. Statistical analysis showed that a significant intergroup difference in craniofacial size between transferred and nontransferred groups as well as a significant inter strain difference among those of the three strains of recipients were observed. Thus, it was disclosed that embryo transfer technique might retard the prenatal development of craniofacial complex of transferred embryo and that the three strains of recipients contributed unequally to the prenatal development of craniofacial complex of transferred embryo through each of their intrauterine environments as a prenatal maternal effect. These results indicated that the intrauterine environment of the recipient played an important role in the prenatal development of the craniofacial complex of the mice embryo. PMID- 8227294 TI - Morphometric analysis of potential maxillary diastemal dental anlagen in three strains of mice. AB - By means of histomorphometric analysis of temporo-spatial changes of the thickness of oral maxillary epithelium in day 11 and 12 ICR, Swiss, and DBA/2 x B6 mouse embryos, the transitory existence of two potential tooth anlagen was documented. The mesial primordium appeared slightly later and disappeared earlier than the distal one. Their development culminated at dental lamina and bud stage, respectively. These potential vestigial tooth anlagen might correspond to the third and fourth upper premolars present in some recent rodent families and in Paramyidae, the supposed ancestors of actual rodents. PMID- 8227295 TI - Denervated chicken breast muscle displays discoordinate regulation and differential patterns of expression of alpha f and beta tropomyosin genes. AB - The expression of the alpha fast (alpha f) and beta tropomyosin (TM) genes has been analysed with muscle-specific and common cDNA probes after unilateral nerve section of the pectoralis major muscle (PM) in 4-week-old chickens. The following were observed in denervated muscles. (1) The beta TM mRNA, which was repressed during development, reaccumulates in a biphasic curve with the increase in the beta TM protein lagging behind the changes in its mRNA. Accordingly, no beta TM is seen in products translated in vitro from total and polyA+ RNA obtained 1 week after denervation. No such translation block is seen with RNA obtained from control or muscles denervated for 6 weeks. (2) No changes in the alpha fTM mRNA and corresponding protein are observed. (3) RNA processing of the two genes is not changed. (4) In the contralateral muscles, transitory increases in alpha f and beta TM mRNAs are observed while the corresponding proteins remain unchanged. Our data suggest that muscle fibres display early and long-term responses to the loss of neural input which might result from a combination of changes produced by regenerative processes and reprogramming of existing fibres. Moreover, in contrast to normal development, no reciprocal changes of alpha f and beta TM expression are seen in denervated muscles. PMID- 8227296 TI - Identification of functioning regulatory sites and a new myosin binding site in the C-terminal 288 amino acids of caldesmon expressed from a human clone. AB - A partial clone of caldesmon, coding for the C-terminal 288 amino acids, was isolated from a human fetal liver cDNA library and sequenced. Expression of the clone in Escherichia coli produced a peptide called H1 (M(r) 32,549), which inhibited tropomyosin-enhanced actomyosin Mg(2+)-ATPase activity by 90% with half maximal inhibition at 0.03-0.04 mol H1 per mol actin. The inhibition could be reversed by Ca(2+)-calmodulin. H1 bound actin, Ca(2+)-calmodulin and tropomyosin and smooth muscle myosin with high affinities. This latter finding shows the presence of a second myosin-binding site in caldesmon. This was confirmed in thrombic digests of native sheep aorta and chicken gizzard caldesmon. PMID- 8227297 TI - Filament overlap affects TnC extraction from skinned muscle fibres. AB - Recent studies on calcium regulation of muscle contraction selectively extract troponin C (TnC) from skinned skeletal muscle fibres with a low ionic strength rigor solution containing a Ca2+/Mg2+ chelator. As previous results from this laboratory and others demonstrate a crossbridge effect, especially rigor, on many of the properties of TnC, the effects of filament overlap on TnC extraction from skinned rabbit psoas muscle fibres were investigated. Tension-pCa relationships at a sarcomere length of 2.7 microns were determined before and after a 5 min TnC extraction at sarcomere lengths of 2.3, 2.5, 2.7, 3.1, 3.3 or 3.5 microns with 20 mM Tris, pH 7.8, 5 mM EDTA. The decrease in the post-extraction maximum Ca2+ activated tension, an indicator of the amount of TnC extracted, was linearly related to the overlap of the thick and thin filaments with decreases in tension being associated with a decrease in filament overlap. The smaller fibre diameter at the longer sarcomere length could facilitate diffusion of TnC from fibre segments. However, the wide range of measured diameters, 40-120 microns, accounted for only 14% of the observed tension decrement and shrinking the fibre with polyvinylpyrrolidone did not increase the tension decrement. Increasing the sarcomere length before extraction was also found to decrease the TnC content of fibre segments along with the post-extraction maximum tension. Thus, TnC appears to be preferentially extracted from non-overlap than overlap regions of the sarcomere. These results further indicate that rigor crossbridges affect TnC other than through increased Ca2+ binding and that under the conditions used here, they retard its extraction. PMID- 8227298 TI - Ultrastructural comparison of slack and stretched myotendinous junctions, based on a three-dimensional model of the connecting domain. AB - The vertebrate myotendinous junction contains junctional microfibrils, located in the lamina lucida of the basement membrane. The junctional microfibrils are thought to transmit muscular force across the junctional lamina lucida, also called the connecting domain. If true, deformation of the terminal muscle cell processes and connecting domain during force transmission would be detected as a change in spacing and/or orientation of the junctional microfibrils. This study compared connecting domain morphology in frog semitendinosus muscles fixed in two extremes of resting tension, to elucidate the mechanical properties of the myotendinous junction. An initial study of connecting domain ultrastructure revealed that junctional microfibrils are punctate or spinelike in shape, and that they are distributed in a linear, helically-oriented array on the muscle cell surface. The rows in the surface lattice are 10-15 nm in thickness, have a centre-to-centre distance between rows of approximately 24 nm, and are oriented at approximately 41 degrees with respect of the long axis of the muscle fibre. Comparison of slack and highly stretched myotendinous junctions shows no significant changes in spacing or orientation of either individual junctional microfibrils or rows in the helical surface lattice. Thus, both the connecting domain and terminal cell processes at the myotendinous junction are essentially inextensible under the loading conditions used in this study. PMID- 8227299 TI - Relation between magnetically-applied force and velocity in beads coated with rabbit myosin, sliding on actin cables in Nitellopsis cells. AB - We have succeeded in controlling the sliding movement of myosin-coated magnetizable beads on actin cables in Nitellopsis cells by the inhomogeneous magnetic field adjacent to a small, strong permanent magnet. The relation between magnetic force acting on the bead and the bead velocity was, in many respects, similar to that obtained from the same system by the use of centrifugal force (Oiwa et al., 1990). In particular, force favouring the motion (negative load) had little effect on the velocity until it was sufficient to pull the bead off the actin, whereas a relatively small positive load caused a reduction in velocity to a plateau value. Although the present method does not allow a good control of force direction, it demonstrates the promise of magnetic force in studying in vitro motility. PMID- 8227300 TI - Elastic properties of the titin filament in the Z-line region of vertebrate striated muscle. AB - The characteristics of the titin filament in the vicinity of the Z-line were investigated using immunoelectron microscopy. We used monoclonal titin antibodies T-11 and T-12 on single fibres of frog skeletal muscle, and on Z-line-extracted fibres. It is well established that the I-band region of titin is elastic. We find, however, that the elastic properties are not uniform. The T-12 epitope, which binds near the Z-line at the N1-line level, hardly changes position relative to the Z-line as the sarcomere is stretched. This demonstrates the functional inextensibility of the N1-Z-line region. After extreme stretch (above 6-microns sarcomere length), this zone finally does elongate; thus, the titin molecule in this region is intrinsically elastic. The functional inextensibility seen at shorter sarcomere lengths may, therefore, be a result of binding of titin to the actin filament in the zone near the Z-line. When the Z-line was extracted, the T-12 epitope remained in the same position as in the unextracted fibres; it did not retract from the Z-line. Failure to retract implies that functional anchoring of titin is not exclusive to the Z-line, but includes some site closer to the A-band. Combined with the results of the above-mentioned stretch experiment, this result implies a likely binding of titin to the thin filament either focally at the N1 line or all along the entire N1-Z region. Thus, this region of titin is functionally stiff, but intrinsically elastic. PMID- 8227301 TI - Time course study of the isometric contractile properties of mdx mouse striated muscles. AB - The isometric twitch and tetanic contractions of three hindlimb muscles (soleus, plantaris, extensor digitorum longus) were recorded in situ in groups of mdx and C57BL/10 control mice at young, adult and old ages (3, 4, 6, 8, 13, 26, 39 and 52 weeks). Based on a two-way analysis of variance (age/phenotype) the mdx phenotype did not modify the absolute tension but was associated with a significant decrease in the tetanic tension normalized to muscle weight in all the muscles which became heavier. These results suggest that the contractile material in mdx is not so powerful as in controls. Moreover, significantly faster time to peak and half-relaxation time were observed in mdx soleus and plantaris. Comparison between these contraction characteristics and those of other experimental models suggests that the high percentage of regenerated fibres in mdx muscles could play a role in modifying contractile properties. PMID- 8227302 TI - Structural change of crossbridges of rabbit skeletal muscle during isometric contraction. AB - Structural changes of crossbridges during isometric contraction have been studied by electron microscopy. Chemically skinned rabbit fibres were rapidly frozen either in activating solution or in ATP-free (rigor) solution, freeze-substituted and embedded. Longitudinal sections of muscle fibres show that the number of crossbridges in active fibres (isometric contraction) is approximately the same as in rigor fibres. Crossbridges of the active and rigor states differ in their shapes, angles and manner of arrangement on the thin filaments. In rigor many crossbridges are wide near the thin filaments and narrow near the thick filament shafts; in active fibres they have more uniform width along their length. The angle of the crossbridges in active fibres is somewhat variable. The average angle is approximately 90 degrees to the filament axis. The crossbridges are arranged on the thin filament retaining the 14.3 nm thick filament periodicity. The crossbridges in rigor are tilted and their arrangement near the thin filament reveals the 36 nm actin periodicity. The variability in the shapes of the crossbridges in active fibres is still higher when we look at them in cross sections of muscle fibres. The crossbridge shapes in the cross-sections were classified and the relative frequency of different shapes was determined. The shapes that are commonly observed in active fibres are similar in that the majority of the mass of the crossbridges is farther away from the thin filament than the crossbridges in rigor fibres. PMID- 8227303 TI - Lesions in the hypothalamus after active immunisation against GnRH in the pig. AB - The terminals of the hypothalamic gonadotrophin hormone-releasing hormone (GnRH) neurons are located within the median eminence and thereby extend beyond the protection of the blood-brain barrier. Thus, these terminals may be subjected to direct autoimmune action in animals that are actively immunised against GnRH. Boars (male pigs) (n = 108) were actively immunised against GnRH by two successive injections with synthetic GnRH, covalently coupled to KLH and dissolved in CFA or IFA. They were killed at 26 weeks of age. Immunised boars were selected on the basis of the resultant testes size, which indicates the effectiveness of the immunisation. The hypothalami of 25 selected animals were studied by histological and immunocytochemical techniques and compared with the hypothalami of three sham- and nine control animals. In the immunised animals, changes in the GnRH system had taken place. These comprised dystrophy of the perikarya and a sharp decrease of the GnRH immunocytochemical reactivity in the terminals within the median eminence. In addition, various degrees of inflammatory reactions were present, particularly within the median eminence. These consisted of tissue disruption by edema, collapse of the capillaries, fibrosis and infiltration with fibroblasts. In addition, accumulations of neurosecretum within the median eminence in combination with hypertrophy of magnocellular neurons within the hypothalamus were present. The reactions were restricted to the median eminence and did not involve other neurohemal organs or other parts of the hypothalamus. A correlation could be established between the incidence of the lesions and the effectiveness of the GnRH autoimmunity (as indicated by the size and endocrine function of the gonads and the anti-GnRH titres). Changes in extra- and intracellular IgG immunocytochemical reactivity within the median eminence indicated the involvement of IgG. The effects were absent from control and sham vaccinated animals and after vaccinations with other compositions of the vaccine. Thus, hypothalamic lesions have been observed in this selected group of animals, vaccinated against GnRH with this particular vaccine. PMID- 8227304 TI - Demonstration of interleukin-1 beta in Lewis rat brain during experimental allergic encephalomyelitis by immunocytochemistry at the light and ultrastructural level. AB - Interleukin-1 beta (IL-1) is a cytokine which exerts many biological effects during inflammation. In the present study, experimental allergic encephalomyelitis (EAE) was induced in Lewis rats. During the various stages of EAE, the presence of IL-1 in the brain was investigated using immunocytochemistry at both the light and ultrastructural level. Ten days after immunization, IL-1 immunoreactivity was found in brains of animals which at this time showed mild clinical signs. Outside the blood-brain barrier, IL-1 was localized in the cytoplasm of meningeal macrophages and perivascular cells. Within the brain parenchyma, IL-1 immunoreactivity was distributed in perivascular lesions in the cytoplasm of infiltrated macrophages and activated microglia. On day 13, animals had developed a full blown EAE. At this stage the number of lesions with IL-1 positive cells had increased. In the remission phase (day 25), lesions with IL-1 positive cells could still be detected but were less pronounced as compared to day 13. Other presumptive IL-1-producing cell types like endothelial cells or astrocytes were, at none of the various stages, found to stain for IL-1. PMID- 8227305 TI - Immune activation in myasthenia gravis: soluble interleukin-2 receptor, interferon-gamma and tumor necrosis factor-alpha levels in patients' serum. AB - Soluble interleukin-2 receptor (sIL-2R), interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) levels were evaluated in serum from patients affected by myasthenia gravis (MG). sIL-2R titers were significantly increased in generalized and bulbar MG patients while ocular cases were not different from controls. Patients showing a recent clinical worsening had significantly higher sIL-2R titers when compared to the whole MG population. sIL-2R levels did not correlate with the corresponding anti-acetylcholine receptor antibody titer (Anti AChR Ab). IFN-gamma was not detected in serum of both MG patients and healthy subjects while TNF-alpha levels were not statistically different from controls. The finding of increased sIL-2R levels supports the hypothesis of circulating activated autoreactive T cells in myasthenic patients. PMID- 8227306 TI - Virus specificity and isotype expression of intraparenchymal antibody-secreting cells during Sindbis virus encephalitis in mice. AB - To study the generation of specific antibody responses within the central nervous system (CNS), we have utilized a murine model of acute viral encephalitis. When Sindbis virus (SV) is injected intracerebrally into weanling mice it causes an acute non-fatal encephalitis and recovery is primarily dependent on the development of antiviral antibody. We used a modified enzyme-linked immunoassay to determine the number of antibody-secreting cells (ASC) specific for SV and their Ig isotype in brain, spleen and cervical lymph nodes over the course of the acute encephalitis. The numbers of SV-specific ASC peak early in spleen and lymph nodes and then begin to increase in brain, suggesting that initial stimulation of B cells occurs primarily in peripheral lymphoid tissue followed by B cell entry into the circulation and appearance in the brain. The pattern for each individual isotype was similar with peak numbers of SV-specific cells present in the spleen 5-7 days after infection, while numbers in the brain continue to rise through day 20 when most ASC were secreting IgG2a or IgA SV-specific antibody. The data suggest therefore that most isotype switching from IgM to IgG and IgA occurs in peripheral lymphoid tissue. An exception to this pattern is IgG1, where numbers of ASC producing IgG1 do not show a peak in spleen and continue to rise in brain through the course of acute encephalitis. The data also indicate that early in infection a large proportion of ASC in the brain are not specific for SV and demonstrate that recruitment of ASC into the CNS is non-specific.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227307 TI - Immunological analysis of the rats with anterior hypothalamic lesions. AB - We have previously identified the suppression of lymphocyte blastogenesis and the acceleration of growth of subcutaneous tumors by bilateral anterior hypothalamic lesions in rats. The present study was performed to clarify the changes in lymphocyte subsets and natural killer (NK) activity after making the lesions. The influence on immunological memory was also studied. The CD4/CD8 ratio of peripheral blood lymphocytes and spleen cells, T cell receptor alpha beta positive cells of the thymocytes and NK activity of the spleen cells decreased significantly. Major histocompatibility complex (MHC) antigen expression on RBL-1 cells injected intraperitoneally into pre-immunized rats was also suppressed. These results suggest that the anterior hypothalamus has some influences on the control of the cellular immunological functions at the peripheral level, on the maturation of T cells at the level of thymus and on the antigen recognition by T cell receptors and MHC antigens. PMID- 8227308 TI - Reactivity of a human monoclonal anti-GM1 and anti-GD1b IgM antibody with human neurons in cultures. AB - A serum containing a monoclonal IgM lambda with anti-GM1 and anti-GD1b activity was obtained from a patient with upper motor neuron syndrome. By indirect immunocytochemical techniques with double staining, the patient's IgM strongly stained membranes of neurons in primary cultures of fetal central and peripheral nervous system. It was cytotoxic for neurons in two human neuroblastoma established cell lines in a complement-dependent chromium release assay. These results are in keeping with the hypothesis of a direct pathogenetic role of such monoclonal anti-GM1 and GD1b IgM antibodies. PMID- 8227309 TI - Identification of Fc gamma RI, II and III on normal human brain ramified microglia and on microglia in senile plaques in Alzheimer's disease. AB - Using monoclonal antibodies to the three known human leukocyte IgG receptors, Fc gamma R, we examined the expression of Fc gamma R in normal brains and in Alzheimer's disease. We found Fc gamma RI, II and III immunoreactivity in senile plaques and on ramified microglia throughout the cortex and white matter of normal and Alzheimer's disease brains. Fc gamma RI expression was independently confirmed by a murine isotype binding study. These findings suggest that intrinsic Fc gamma R may play an important role in normal and disordered immune related processes in the brain. They support the idea that microglia are brain macrophages. PMID- 8227310 TI - Beta-endorphin protects mice from neurological disease induced by the murine coronavirus MHV-JHM. AB - The neurotropic murine coronavirus, MHV-JHM (JHMV) causes encephalitis and paralytic-demyelinating disease in susceptible strains of mice and rats, serving as a model for human demyelinating diseases such as multiple sclerosis. In this communication, we report that a single intracerebral administration of the naturally occurring neuropeptide, beta-endorphin, reduced the incidence of JHMV induced paralytic-demyelinating disease 40-50% in C57Bl/6 mice. Protection from disease was accompanied by significantly reduced virus replication in the brain as early as 3 days post-infection and did not occur in irradiated, or immunoincompetent mice. The data suggest that beta-endorphin engages immune mechanisms of host resistance to JHMV infection to protect the mice from disease. PMID- 8227311 TI - Correlation between susceptibility to demyelination and interferon-gamma induction of major histocompatibility complex class II antigens on murine cerebrovascular endothelial cells. AB - The induction of major histocompatibility complex (MHC) Class II expression was studied on cerebrovascular endothelial cells (CVE) obtained from strains of mice that are resistant (BALB/c) and susceptible (SJL and CBA) to Theiler's virus induced demyelination (TVID). Following 24 h treatment with interferon (IFN) gamma, MHC Class II was induced on CVE derived from susceptible but not resistant strains of mice. However, IFN-gamma induced the expression of MHC Class II on late passages of BALB/c CVE. These results demonstrate a correlation between susceptibility to demyelination and the ability of IFN-gamma to induce the expression of MHC Class II on CVE. In susceptible strains of mice, the presence of activated, IFN-gamma-secreting T cells, in the vicinity of CVE would increase the antigen-presenting capabilities of CVE and result in increased T cell traffic into the central nervous system. PMID- 8227312 TI - Stimulation of natural killer and antibody-dependent cellular cytotoxicity activities in mouse leukocytes by bombesin, gastrin-releasing peptide and neuromedin C: involvement of cyclic AMP, inositol 1,4,5-trisphosphate and protein kinase C. AB - Bombesin and the two mammalian bombesin-related peptides, gastrin-releasing peptide (GRP) and neuromedin C, at physiological concentrations ranging from 10( 11) M to 10(-9) M have been shown in this study to significantly stimulate in vitro the antibody-dependent cellular cytotoxicity (ADCC) and natural killer (NK) activities in BALB/c mouse leukocytes from axillary nodes, spleen and thymus. The three neuropeptides studied induced no change in interleukin-2 production. In addition, these neuropeptides induced in leukocytes from axillary nodes a rapid, transient and significant decrease of intracellular cyclic AMP at 30 s, but a significant transient increase of inositol 1,4,5-trisphosphate levels at 30 and 60 s and a stimulation of protein kinase C activity in membrane fractions after 5 min incubation. These results suggest that inositol phospholipid signalling and cAMP messenger systems are involved in the increase of NK and ADCC activities when leukocytes are incubated in the presence of bombesin, GRP or neuromedin C. PMID- 8227313 TI - Mechanisms of stress-induced modulation of viral pathogenesis and immunity. AB - A murine model of herpes simplex virus (HSV) infection was used to examine the roles of catecholamines and corticosterone in the restraint stress-induced suppression of viral immunity. Treatment of C57BL/6 mice with RU486, a glucocorticoid receptor antagonist, reversed the stress-induced diminution of cellularity in response to local HSV infection. Treatment of mice with both nadolol, a peripherally acting beta-adrenergic antagonist, and RU486 completely reversed the restraint stress-induced suppression of HSV-specific CTL activation. These findings demonstrate that both corticosterone and catecholamine-mediated mechanisms are operative in the stress-induced suppression of anti-viral cellular immunity. PMID- 8227314 TI - Lymphocyte adhesion to cultured endothelial cells of the blood-retinal barrier. AB - Microvascular endothelial cells derived from the blood-retinal barrier were grown in vitro and various factors affecting the adhesion of syngeneic lymphocytes to these monolayers was evaluated. Under resting conditions 5.3 +/- 0.4% of lymphocytes derived from peripheral lymph nodes (PLN) were found to adhere to the endothelia. Adhesion of resting lymphocytes increased significantly following endothelial treatment with interferon-gamma (IFN-gamma; 11.7 +/- 1.0%), interleukin-1 (IL-1; 14.9 +/- 1.2%), astrocyte conditioned medium (ACM; 12.7 +/- 0.9%) or forskolin (13.9 +/- 1.2%). Lymphocyte activation with concanavalin A (ConA) increased adhesion to 17.0 +/- 0.9% which could be augmented by activating the endothelia with IFN-gamma (22.3 +/- 1.0%), IL-1 (24.0 +/- 1.0%) and ACM (25.7 +/- 1.6%). An antigen-specific CD4+ T cell line exhibited the greatest degree of adhesion, 40.4 +/- 2.5% on resting endothelia, 60.0 +/- 3.0% on IFN-gamma activated cells and 54.3 +/- 1.4% on IL-1-activated cells. Although CD4+ lymphocytes predominated in the PLN population by 2:1, significantly more CD8+ cells were found to adhere. PMID- 8227315 TI - Ciliary neurotrophic factor (CNTF) promotes low-affinity nerve growth factor receptor and CD4 expression by rat CNS microglia. AB - Ramified parenchymal microglia may provide immune surveillance in the nervous system and become activated in response to injury, showing increases in antigens found on macrophages, e.g. CD4 and MHCs. We investigated in adult rats the effects of a 2-week intraventricular infusion with ciliary neurotrophic factor (CNTF), a nervous system-associated cytokine, on microglia of the normal and injured corpus callosum. CNTF caused morphological changes, induced the expression of low-affinity nerve growth factor receptor and CD4 and increased the expression of complement receptor 3. Such changes were also observed after treatment of pure cultures of neonatal rat microglial cells with highly purified CNTF, suggesting a direct responsiveness to CNTF. Thus, endogenous astroglial and Schwann cell-derived CNTF may be an important component of the immune responses of the nervous system. PMID- 8227316 TI - Microglia in the immune surveillance of the brain: human microglia constitutively express HLA-DR molecules. AB - The degree of MHC class II expression in histologically normal human brain biopsy or autopsy tissue is still controversial. According to the generally held view MHC class II expression is rather low in the normal brain with the exception of the white matter. In the present study, HLA-DR expression was examined immunocytochemically in different brain areas obtained from three autopsy cases with short post-mortem times (i.e. 6 h). Based on standard histological evaluation, the brain areas studied appeared as histologically normal tissue. In all brain areas there was a strong constitutive HLA-DR expression on ramified microglia. The number of HLA-DR-immunoreactive microglia was strongest in the white matter (the corpus callosum and the capsula interna for example). The border zone between white matter and grey matter, however, revealed a sharp contrast between a high density of HLA-DR-immunoreactive microglia in the white matter and a rather low number in the grey matter. In the grey matter, HLA-DR immunoreactive microglia were much less frequent than in the white matter and more pronounced on perivascular cells. The staining and distribution pattern of HLA-DR-immunoreactive microglia was confirmed by immunocytochemistry with a panel of different anti-HLA-DR monoclonal antibodies as well as by quantitative analysis of the immunostaining. Unlike the HLA-DR immunoreactivity, HLA-ABC immunoreactivity (detecting MHC class I antigens) was confined to endothelia and not observed on microglia. In the choroid plexus stromal macrophages expressed both class I and II antigens (i.e. at a location which could provide the peripheral immune system access to CNS antigens). Constitutive HLA-DR expression by microglia qualifies them as the main resident antigen-presenting cell of the brain. The pronounced overall HLA-DR expression by resting microglia questions a central dogma of the brain as an immune-privileged site and further points to the key role of the microglia in brain immune surveillance. PMID- 8227317 TI - Binding of IgG to amyloid beta A4 peptide via the heavy-chain hinge region with preservation of antigen binding. AB - Amyloid beta A4 peptide is found in the extracellular region of the senile plaque and in the angiopathy of Alzheimer's disease. Several other proteins, including IgG, also reside in these abnormal structures. In an attempt to understand how these structures are assembled and to determine how proteins are recruited, interactions of various proteins with synthetic beta A4 peptide have been examined in vitro. Purified IgG binds directly to synthetic beta A4 peptide with high avidity. The domain between amino acids 12-28 of beta A4 binds IgG. beta A4 peptide binds the hinge region of the immunoglobulin heavy chain, and preserves the ability of the immunoglobulin to bind antigen. A protein which does not bind directly to beta A4 peptide can be targetted to the senile plaque and angiopathy by binding to IgG, which avidly binds beta A4 peptide. PMID- 8227318 TI - Endogenous gamma interferon produced in central nervous system by systemic infection with Theiler's virus in mice. AB - Theiler's virus GD VII strain causes acute encephalomyelitis by intracerebral inoculation. We established acute encephalomyelitis in mice by the intravenous (i.v.) inoculation of Theiler's virus GD VII strain. Replication of Theiler's virus injected i.v. could be observed in both the brain and spinal cord of mice, and interferon (IFN)-gamma could be detected in the extracts of brain and spinal cord in parallel with viral replication. Furthermore, by the injection of anti IFN-gamma monoclonal antibody (mAb) on Day 1 post-infection (p.i.), mortality and virus titres in the spinal cord increased compared with the control mice treated with normal rat globulin. The histological exacerbation of inflammation was observed in spinal cord of anti-IFN-gamma mAb-treated mice. These results indicate that endogenous IFN-gamma, produced locally in the brain and spinal cord of mice through both antiviral action and anti-inflammatory action of IFN-gamma in central nervous system, plays an important role in Theiler's virus infection. PMID- 8227319 TI - Encephalitogenic Th1 cells are inhibited by Th2 cells with related peptide specificity: relative roles of interleukin (IL)-4 and IL-10. AB - Cytokines secreted by T-helper type 2 (Th2) cells inhibit the antigen-induced stimulation of type 1 (Th1) helper T cells. To study this form of regulation in an autoimmune disease model, the cytokines secreted by a Th2 clone specific for the encephalitogenic proteolipid protein (PLP) peptide 139-151 were tested for their ability to inhibit proliferation of an encephalitogenic Th1 clone specific for an epitope contained within the same peptide. Cytokines, produced by stimulation of the Th2 clone with CD3-specific monoclonal antibodies (mAbs), inhibited proliferation of the Th1 clone when stimulated by antigen and splenic antigen-presenting cells (APC). Inhibition was, however, not antigen-specific since cytokines released upon stimulation of an unrelated Th2 clone were also inhibitory. Inhibition was found to be caused by effects on either antigen presentation or co-stimulatory activity of the APC and not by direct effects on the Th1 cells. MAbs for the two major regulatory Th2 cytokines were used to identify the inhibitory component secreted by activated Th2 cells. Interleukin-10 (IL-10)-specific mAb abolished the inhibitory effect, while mAb specific for IL-4 had no effect on inhibition. The addition of recombinant IL-4 (rIL-4) and rIL-10 confirmed that inhibition of Th1 proliferation was due to secretion of IL-10 by the Th2 clone and its subsequent effects on APC. The studies described here demonstrate that PLP-specific Th2 cells which recognize peptide 139-151 inhibit encephalitogenic Th1 cells which respond to an epitope on the same peptide. This phenomenon may be important for local, antigen-specific regulation of inflammation in the central nervous system. PMID- 8227320 TI - Low levels of specific T cell activation marker CD27 accompanied by elevated levels of markers for non-specific immune activation in the cerebrospinal fluid of patients with AIDS dementia complex. AB - Concentrations of soluble receptors for tumor necrosis factor (sTNFR-p55 and sTNFR-p75) and soluble T cell antigens CD25 and CD27 (sCD25 and sCD27) were measured in paired serum/cerebrospinal fluid (CSF) samples of 15 patients with AIDS dementia complex (ADC) and 15 HIV-infected control subjects (11 with other central nervous system (CNS) infections and four without CNS infection). In this study levels of sTNFR-p55, sTNFR-p75 and sCD25 were elevated in the CSF of ADC patients and of the 11 patients with other CNS infections, whereas CSF-levels of the specific T cell marker sCD27 were lower in patients with ADC as compared to the control subjects with and without other CNS infections. This pattern suggests a relative failure of eliciting a T cell-mediated immune response intrathecally in patients with ADC. PMID- 8227321 TI - Diseased G protein-coupled receptors. PMID- 8227322 TI - Protease nexin-2/amyloid beta-protein precursor: a cerebral anticoagulant? PMID- 8227323 TI - Bile salt-induced diarrhea: the cellular mechanism. PMID- 8227324 TI - Glucokinase as pancreatic beta cell glucose sensor and diabetes gene. PMID- 8227325 TI - Tissue-specific expression of the gene coding for human Clara cell 10-kD protein, a phospholipase A2-inhibitory protein. AB - Clara cell 10-kD protein (cc10kD), a secretory phospholipase A2 inhibitor, is suggested to be the human counterpart of rabbit uteroglobin (UG). Because cc10kD is expressed constitutively at a very high level in the human respiratory epithelium, the 5' region of its gene may be useful in achieving organ-specific expression of recombinant DNA in gene therapy of diseases such as cystic fibrosis. However, it is important to establish the tissue-specific expression of this gene before designing gene transfer experiments. Since the UG gene in the rabbit is expressed in many other organs besides the lung and the endometrium, we investigated the organ and tissue specificity of human cc10kD gene expression using polymerase chain reaction, nucleotide sequence analysis, immunofluorescence, and Northern blotting. Our results indicate that, in addition to the lung, cc10kD is expressed in several nonrespiratory organs, with a distribution pattern very similar, if not identical, to that of UG in the rabbit. These results underscore the necessity for more detailed analyses of the 5' region of the human cc10kD gene before its usefulness in gene therapy could be fully assessed. These data also suggest that cc10kD and UG may have similar physiological function(s). PMID- 8227326 TI - A reporter transgene indicates renal-specific induction of tumor necrosis factor (TNF) by shiga-like toxin. Possible involvement of TNF in hemolytic uremic syndrome. AB - We have examined the hypothesis that TNF may play a pathogenetically important role in the hemolytic uremic syndrome. Specifically, we considered the possibility that shigatoxin, which eventuates this syndrome, might induce TNF biosynthesis, and/or that TNF and shigatoxin might sensitize animals, each to the toxic effects of the other agent. Shigatoxin was found to sensitize mice to the lethal effect of LPS and to the lethal effect of TNF. On the other hand, pretreatment of animals with either TNF or LPS did not noticeably sensitize mice to the lethal effect of shigatoxin. Intraperitoneal injections of shigatoxin did not induce the production of detectable quantities of TNF in the plasma of mice. When shigatoxin was injected into transgenic mice bearing a chloramphenicol acetyltransferase (CAT) reporter gene that indicates TNF synthesis, CAT activity was induced within the kidney, but not in other tissues. We therefore conclude that shigatoxin acts to induce TNF synthesis within the kidney, and at the same time increases renal sensitivity to the toxic effects of TNF. While this mouse model does not reproduce the hemolytic uremic syndrome as it occurs in humans, it does suggest that local synthesis of TNF within the kidney may contribute to renal injury induced by shigatoxin. PMID- 8227327 TI - Antigen mimicry in autoimmune disease sharing of amino acid residues critical for pathogenic T cell activation. AB - A nonamer peptide from murine nicotinic acetylcholine receptor delta chain (ACR delta), which shared four amino acid residues with a nonamer peptide of murine ovarian zona pellucida glycoprotein ZP3, induced murine autoimmune oophoritis and IgG autoantibody to the zona pellucida. Crossreaction between the ACR delta and ZP3 peptides was established by the response of a ZP3 peptide-specific, oophoritogenic T cell clone to both peptides in association with IA (alpha k beta b). By substituting the ZP3 peptides with a single alanine, four amino acids within the ZP3 peptide were found to be important for ovarian autoimmune disease, autoantibody response, and stimulation of the ZP3-specific T cell clone. Substitution with conservative amino acids of three residues also ablated activity, whereas the fourth, a phenylalanine, was replaceable by tyrosine without loss of activity. Of the four critical amino acids, three were shared between the ZP3 peptide and the ACR delta peptide. Moreover, polyalanine peptides with the four critical ZP3 amino acids or the four amino acids common to the ZP3 and ACR delta peptides induced immune response to ZP3 and elicited severe ovarian autoimmune disease. Thus, organ-specific autoimmune disease can occur through immune response against unrelated self (or foreign) peptides that share with a self-peptide sufficient common amino acid residues critical for activation of pathogenic, autoreactive T cells. PMID- 8227328 TI - The regulation of adipose tissue and muscle lipoprotein lipase in runners by detraining. AB - To study the mechanism of lipoprotein lipase (LPL) regulation by exercise, we recruited 16 healthy athletes to undergo a 2-wk period of detraining. Fasting fat and muscle biopsies were performed both before and after the detraining period. In muscle, detraining resulted in a decrease in LPL activity in both the heparin releasable (HR) (-45%, P < 0.05) and cellular (extractable [EXT]) (-75%, P < 0.005) fractions, with no significant changes in LPL immunoreactive mass and mRNA levels. However, several subjects demonstrated parallel decreases in LPL mass and mRNA levels with detraining, suggesting that there is some degree of heterogeneity in response. In adipose tissue, detraining had the opposite effects on LPL activity. In the HR fraction, detraining resulted in an 86% increase (P < 0.005) in LPL activity, which was paralleled by a 100% (P = 0.02) increase in HR mass. However, there was no significant change in EXT LPL activity or EXT LPL mass. There were no changes in adipose LPL synthetic rate or LPL mRNA levels with detraining. The ratio of adipose tissue/muscle LPL, which may be an important indicator of the tendency for storage of circulating lipids in adipose tissue, increased significantly after detraining. The adipose/muscle LPL ratio was 0.51 +/- 0.17 in the exercising runners, and 4.45 +/- 2.46 in the same runners after detraining (P < 0.05). Thus, detraining of athletes resulted in a decrease in muscle LPL that occurred through post-translational mechanisms, whereas adipose tissue LPL increased, also due to posttranslational changes. This decrease in muscle LPL, coupled with an increase in adipose LPL, yielded a condition favoring adipose tissue storage. PMID- 8227329 TI - Mechanism of the anticoagulant effect of warfarin as evaluated in rabbits by selective depression of individual procoagulant vitamin K-dependent clotting factors. AB - We have evaluated the contribution of depression of individual procoagulant vitamin K-dependent clotting factors to the ability of warfarin to protect rabbits against tissue factor-induced coagulation. Mean activities of individual procoagulant factors were determined, in assays with rabbit substrates, for a group of rabbits achieving a protective degree of anticoagulation with warfarin. Values were: factor VII, 12%; factor IX, 7%; factor X, 14%, and prothrombin, 13%. The effect upon tissue factor-induced coagulation of selective immunodepletion of each factor to a comparable level was then evaluated. Immunodepletion of plasma factor X or prothrombin, but not of factor VII or factor IX, protected otherwise normal rabbits against tissue factor-induced coagulation. Next, we determined the effect upon the protection in warfarin-treated rabbits of selectively restoring factor X or prothrombin before infusing tissue factor. When either factor was selectively restored, warfarin's protective effect was abolished. Moreover, selective restoration of prothrombin sensitized warfarin-treated rabbits to coagulation more severe than observed in nontreated control rabbits. One may extrapolate from these data that depression of both factor X and prothrombin are required for warfarin's clinical antithrombotic efficacy and that depression of plasma prothrombin is particularly important. PMID- 8227330 TI - Suramin interferes with interleukin-6 receptor binding in vitro and inhibits colon-26-mediated experimental cancer cachexia in vivo. AB - Neoplastic diseases are frequently associated with metabolic changes collectively known as cancer cachexia. The presence of cachexia complicates therapeutic intervention and is an important cause of death in cancer patients. At present there is no effective treatment for cachexia. Recently, the involvement of interleukin-6 (IL-6) in the wasting of colon-26 adenocarcinoma-bearing mice was demonstrated. The research presented here establishes an anticachectic role for the experimental drug suramin, since it partially blocks (up to 60%) the catabolic effects associated with the growth of this tumor in vivo. Suramin prevents the binding of IL-6 to its cell surface receptor subunits, as demonstrated by radioreceptor binding assay and affinity crosslinking experiments. Furthermore, the uptake of radioactive IL-6 by the liver is significantly reduced in suramin-treated mice. On the other hand, the drug is approximately 10-fold less potent in inhibiting the binding of tumor necrosis factor-alpha to indicator cell line in vitro and fails to block liver uptake of this cytokine in vivo. Collectively, these results suggest that suramin inhibits cancer-associated wasting, in part by interfering with the binding of IL-6 to its receptor. Whether suramin inhibits the action of other factors/cytokines that may also participate in colon-26-mediated cachexia is not yet known. PMID- 8227331 TI - A ligand-free, soluble urokinase receptor is present in the ascitic fluid from patients with ovarian cancer. AB - We have identified a soluble form of the human urokinase plasminogen activator (uPA) receptor (uPAR) in the ascitic fluids from patients with ovarian cancer. After purification of uPAR from the ascitic fluids by ligand-affinity chromatography (pro-uPA Sepharose), the uPAR was initially identified by cross linking to a radiolabeled amino-terminal fragment of human uPA. The uPAR purified from the ascitic fluid has no bound ligand (uPA), as similar amounts can be purified by ligand-affinity chromatography as by immuno-affinity chromatography. uPAR from ascitic fluids partitions in the water phase after a temperature dependent phase separation of a detergent extract. It therefore lacks at least the lipid moiety of the glycophospholipid anchor present in cellular-bound uPARs. It is highly glycosylated and the deglycosylated form has the same electrophoretic mobility as previously characterized cellular uPAR from other sources. The immunoreactivity of the purified uPAR from the ascitic fluid is indistinguishable from that of characterized uPAR, demonstrated by Western blotting with three different anti-uPAR monoclonal antibodies. The uPAR was found in 11 of 11 ascitic fluids from patients with ovarian cancer and in elevated amounts in the plasma from 2 of 3 patients. The concentration of soluble uPAR in the ascitic fluid was estimated to range between 1 and 10 ng/ml. Human soluble uPAR, derived from the tumor cells, was also found in the ascitic fluid and serum from nude mice xenografted intraperitoneally with three different human ovarian carcinomas. PMID- 8227332 TI - The G proteins of the G alpha i and G alpha q family couple the bradykinin receptor to the release of endothelium-derived relaxing factor. AB - Bradykinin stimulates diverse functions in endothelial cells including the release of endothelium-derived relaxing factor (EDRF). Little is known, however, regarding the identity of the G protein(s) involved. Here we demonstrate that G proteins of the G alpha i and G alpha q family are coupled to the bradykinin receptor (BKR) in bovine aortic endothelial cells by using specific antisera directed against the COOH-terminal region of G alpha i2 (P4), G alpha i3 (EC), and G alpha q (QL). These antisera are specific since their effects are blocked by the decapeptides from which they were derived. The degree of receptor-G protein coupling was assessed by the formation of high affinity agonist binding sites (HABS) and GTP hydrolysis. In a concentration-dependent manner, the QL antisera reduced HABS and GTPase activity by 65 and 60%, respectively, and effectively abolished them in membranes from pertussis toxin-treated cells. The combination of P4 and EC antisera produced a loss of HABS (41%) and GTPase activity (40%) comparable to the effects of pertussis toxin. These findings indicate that G alpha i and G alpha q proteins mediate the cellular responses to bradykinin in bovine aortic endothelial cells and support the observation that bradykinin-stimulated EDRF release is relatively insensitive to pertussis toxin. PMID- 8227333 TI - Molecular analysis of patient and carrier genes with congenital steroid 21 hydroxylase deficiency by using polymerase chain reaction and single strand conformation polymorphism. AB - Steroid 21-hydroxylase deficiency is a major cause of congenital adrenal hyperplasia and is caused by genetic impairment of this enzyme. Since approximately 80% of cases are caused by point mutations of the CYP21B (CYP21A2) gene, whereas the remaining 20% are due to deletion of this gene, we used the polymerase chain reaction single strand conformation polymorphism technique for rapid and accurate diagnosis of this disease. Of 23 patients examined, 1 had a hemizygous CYP21B gene. 18 patient's genes localized their harmful mutations or deletion on both the alleles, while 4 of them found their causative mutations on one of the two alleles, and 1 failed to find any responsible mutation. All the mutations (four nucleotide substitutions) detected are also found in the CYP21A (CYP21A1) pseudogene. A mutation at the intron 2 site is most prevalent in both salt-wasting and simple virilizing forms of the disease, and accounts for 37% of the patient's genes (17/46). Pedigree analysis of these mutations revealed that the mutations (at least four of them) occurred de novo at a considerable frequency on both the paternally and maternally inherited chromosomes. This result could explain occasional discordance of the diagnosis using HLA typing with the clinical symptoms. PMID- 8227334 TI - Lipoprotein lipase regulation by insulin and glucocorticoid in subcutaneous and omental adipose tissues of obese women and men. AB - There are marked variations in the activity of lipoprotein lipase (LPL) among adipose depots, particularly in women. Consistent with data on LPL activity, the level of expression of LPL mRNA was lower in omental (OM) than subcutaneous (SQ) adipose tissue of women. To investigate the cellular basis of these differences, OM and SQ adipose tissues obtained at surgery from obese men and women were placed in organ culture for 7 d with varying concentrations of insulin and dexamethasone. Insulin increased levels of LPL mRNA and LPL activity in abdominal SQ but not OM adipose tissue. Dexamethasone also increased LPL mRNA and LPL activity, and these effects were more marked in the OM adipose tissue, particularly in men. When insulin and dexamethasone were added together, synergistic increases in LPL activity were seen in both depots, and this was in part explained at the level of LPL mRNA. The SQ depot was more sensitive to the effects of submaximal doses of dexamethasone in the presence of insulin. The maximum activity of LPL induced by insulin or insulin plus dexamethasone was higher in the SQ than in the OM depot of women, and this was associated with higher levels of LPL mRNA. Rates of LPL synthesis paralleled LPL mRNA levels. These data show that insulin and glucocorticoids influence human adipose tissue LPL activity at the level of LPL gene expression, as well as posttranslationally, and that responsiveness to these hormonal effects is dependent on adipose depot and gender. PMID- 8227335 TI - Early events of human T lymphocyte activation are associated with type I protein kinase A activity. AB - Human T lymphocytes possess both the type I and II isozymes of protein kinase A (PKA). The type I (PKA-I) isozyme is predominantly associated with the plasma membrane, whereas the type II (PKA-II) isozyme is primarily localized to the cytosol. Because the functions of both PKA-I and PKA-II isozymes in the biochemical events of T lymphocyte activation have not been clearly elucidated, we tested the hypothesis that very early events of normal human T lymphocyte activation are mediated by the PKA-I and/or PKA-II isozyme(s). Fresh normal human T cells and a normal human CD4+ T cell line (GK606) activated with anti-CD3 epsilon and recombinant interleukin 1 alpha (rIL-1 alpha) exhibited a peak six- to sevenfold increase of PKA phosphotransferase activity at 5 min that returned to baseline by 60 min. Similarly, both fresh T cells and the T cell line activated by phorbol myristate acetate and ionomycin demonstrated a peak eightfold increase of PKA activity by 15 min that returned toward baseline by 60 min. Chromatographic separation of the PKA isozymes and quantification of phosphotransferase activities after T cell activation by either agonist pair showed preferential activation of the PKA-I isozyme, resulting in a significant reduction in the ratio of PKA-I to PKA-II isozyme activity from 3.1:1-6.2:1 to 1.1:1-3.2:1. PKA-I isozyme activation resulted in the release of free catalytic (C) subunit, an increase in C subunit phosphotransferase activity, and the phosphorylation of T cell plasma membrane-associated proteins, p14, p17, p20, p21, p38, and p48. However, activation of the PKA-I isozyme did not appear to be required for the transcription of IL-2 mRNA, an event necessary for mitosis. These data indicate that ligand-induced T cell activation is associated with rapid activation of the PKA-I, but not PKA-II, isozyme that results in phosphorylation of plasma membrane-associated proteins. The involvement of the PKA-I isozyme during the very early events of T cell activation suggests that this isozyme may be an antigen- or mitogen-stimulated protein kinase. PMID- 8227336 TI - Transport of beta-lactam antibiotics in kidney brush border membrane. Determinants of their affinity for the oligopeptide/H+ symporter. AB - This study was designed to determine whether beta-lactam antibiotics (cephalosporins and penicillins) are all substrates for the renal oligopeptide/H+ symporter and, if so, whether the transport system discriminates among the numerous beta-lactam antibiotics. We used [3H]glycylglutamine, [3H]cephalexin, and [3H]-ampicillin as probes for the transport of oligopeptides, cephalosporins, and penicillins in kidney brush border membrane vesicles, respectively. Among the beta-lactam antibiotics, only those with an alpha-amino group in the phenylacetamido moiety were found to interact with the oligopeptide/H+ symporter. Aminocephalosporins displayed high affinities (KiS generally < 250 microM), whereas aminopenicillins displayed low affinities (Ki 0.78-3.03 mM). These differences in affinities appeared to be a consequence of conformational features of the substrates, especially the sterical location of the carboxy group. The affinities of aminolactams for the oligopeptide/H+ symporter were, furthermore, related to the hydrophobicity of the phenylglycyl chains and the substituents attached to the thiazolidine and dihydrothiazine ring. In sharp contrast to the uptake of [3H]glycylglutamine and [3H]cephalexin, the uptake of [3H]ampicillin was not dependent on a pH gradient and was inhibited by various beta-lactam antibiotics, whether or not they contained an alpha-amino group. Our data suggest that: (a) the transport of aminocephalosporins is largely mediated by the oligopeptide/H+ symporter, which is highly influenced by the substrate structure; and (b) penicillins are transported by another system, which is less discriminative with respect to substrate structure. PMID- 8227337 TI - Protamine-mediated transport of albumin into brain and other organs of the rat. Binding and endocytosis of protamine-albumin complex by microvascular endothelium. AB - High doses of intravenous protamine cause generalized vascular permeability changes in brain and other organs, and concomitant hypoproteinemia. The present investigations test the hypothesis that protamine has a dual action of both binding serum proteins and of undergoing absorptive-mediated transcytosis through microvascular endothelial barriers. Binding of albumin to protamine was demonstrated using equilibrium dialysis, and protamine was shown to selectively augment the uptake of albumin, but not sucrose, in isolated bovine or human brain capillaries. In contrast, the anionic macromolecule, dextran sulfate, resulted in an increased capillary uptake of both albumin and sucrose in vitro. The selective effects of protamine on albumin transport were also documented in vivo using an external organ technique; the intravenous injection of 1.5 mg/kg protamine resulted in a marked and selective influx of albumin into brain, heart, kidney, lung, and liver, and the increased albumin transport exceeded the increased sucrose uptake in some organs by an order of magnitude. The transcytosis of protamine through the cerebral microvascular barrier was documented with an internal carotid artery perfusion technique. In summary, these studies provide evidence for protamine-mediated vectorial transport of albumin through microvascular barriers in brain and other organs. PMID- 8227338 TI - Opioid antagonist diprenorphine microinjected into parabrachial nucleus selectively inhibits vasopressin response to hypovolemic stimuli in the rat. AB - Subcutaneous injection of the potent, nonselective opioid antagonist diprenorphine inhibits the vasopressin response to acute hypovolemia. To determine if this inhibition is due to antagonism of opioid receptors in brain pathways that mediate volume control, we determined the vasopressin response to different stimuli when diprenorphine or other opiates were injected into the cerebral ventricles, the nucleus tractus solitarius (NTS), or the lateral parabrachial nucleus (PBN) of rats. We found that the vasopressin response to hypovolemia was inhibited by injection of diprenorphine into the cerebral ventricles at a dose too low to be effective when given subcutaneously. This response also was inhibited when a 20-fold lower dose of diprenorphine was injected into the PBN but not when it was injected into the NTS. The inhibitory effect of diprenorphine in the PBN was not attributable to a decrease in osmotic or hypovolemic stimulation and did not occur with osmotic or hypotensive stimuli. Injecting the PBN with equimolar doses of the mu antagonist naloxone, the delta antagonist ICI-154,129 or the kappa-1 agonist U-50,488H had no effect on basal or volume-stimulated vasopressin. We conclude that the inhibition of vasopressin by diprenorphine is due partially to action at a novel class of opioid receptors that transmit volume stimuli through the PBN. PMID- 8227339 TI - Myometrial angiotensin II receptor subtypes change during ovine pregnancy. AB - Although regulation of angiotensin II receptor (AT) binding in vascular and uterine smooth muscle is similar in nonpregnant animals, studies suggest it may differ during pregnancy. We, therefore, examined binding characteristics of myometrial AT receptors in nulliparous (n = 7), pregnant (n = 24, 110-139 d of gestation), and postpartum (n = 21, 5 to > or = 130 d) sheep and compared this to vascular receptor binding. We also determined if changes in myometrial binding reflect alterations in receptor subtype. By using plasma membrane preparations from myometrium and medial layer of abdominal aorta, we determined receptor density and affinity employing radioligand binding; myometrial AT receptor subtypes were assessed by inhibiting [125I]-ANG II binding with subtype-specific antagonists. Compared to nulliparous ewes, myometrial AT receptor density fell approximately 90% during pregnancy (1,486 +/- 167 vs. 130 +/- 16 fmol/mg protein) and returned to nulliparous values > or = 4 wk postpartum; vascular binding was unchanged. Nulliparous myometrium expressed predominantly AT2 receptors (AT1/AT2 congruent to 15%/85%), whereas AT1 receptors predominated during pregnancy (AT1/AT2 congruent to 80%/20%). By 5 d postpartum AT1/AT2 congruent to 40%/60%, and > 4 wk postpartum AT2 receptors again predominated (AT1/AT2 congruent to 15%/85%). In studies of ANG II-induced force generation, myometrium from pregnant ewes (n = 10) demonstrated dose-dependent increases in force (P < 0.001), which were inhibited with an AT1 receptor antagonist. Postpartum myometrial responses were less at doses > or = 10(-9) M (P < 0.05) and unaffected by AT2 receptor antagonists. Vascular and myometrial AT receptor binding are differentially regulated during ovine pregnancy, the latter primarily reflecting decreases in AT2 receptor expression. This is the first description of reversible changes in AT receptor subtype in adult mammals. PMID- 8227340 TI - Insulin-like growth factor I stimulates lipid oxidation, reduces protein oxidation, and enhances insulin sensitivity in humans. AB - To elucidate the effects of insulin-like growth factor I (IGF-I) on fuel oxidation and insulin sensitivity, eight healthy subjects were treated with saline and recombinant human (IGF-I (10 micrograms/kg.h) during 5 d in a crossover, randomized fashion, while receiving an isocaloric diet (30 kcal/kg.d) throughout the study period. On the third and fourth treatment days, respectively, an L-arginine stimulation test and an intravenous glucose tolerance test were performed. A euglycemic, hyperinsulinemic clamp combined with indirect calorimetry and a glucose tracer infusion were performed on the fifth treatment day. IGF-I treatment led to reduced fasting and stimulated (glucose and/or L arginine) insulin and growth hormone secretion. Basal and stimulated glucagon secretion remained unchanged. Intravenous glucose tolerance was unaltered despite reduced insulin secretion. Resting energy expenditure and lipid oxidation were both elevated, while protein oxidation was reduced, and glucose turnover rates were unaltered on the fifth treatment day with IGF-I as compared to the control period. Enhanced lipolysis was reflected by elevated circulating free fatty acids. Moreover, insulin-stimulated oxidative and nonoxidative glucose disposal (i.e., insulin sensitivity) were enhanced during IGF-I treatment. Thus, IGF-I treatment leads to marked changes in lipid and protein oxidation, whereas, at the dose used, carbohydrate metabolism remains unaltered in the face of reduced insulin levels and enhanced insulin sensitivity. PMID- 8227341 TI - Bumetanide decreases canine cerebrospinal fluid production. In vivo evidence for NaCl cotransport in the central nervous system. AB - Na/K/2Cl cotransport carrier plays an important role in fluid absorption and secretion in many epithelial tissues. The role of the carrier, however, in mammalian choroidal cerebrospinal fluid (CSF) production has been controversial. We used ventriculo-cisternal perfusion (VCP) labeled with blue dextran with or without bumetanide and measured choroidal CSF production in anesthetized, and paralyzed, mechanically ventilated dogs. During 3 h of VCP, mean intracerebroventricular and arterial pressures, PaCO2, pH, [HCO3-], and serum osmolality remained normal in both groups (n = 9 in each group). Beginning 90 min after the start of VCP, choroidal CSF production was measured every 15 min. In group I (control group), values for CSF production (means +/- SD) were 49 +/- 20, 49 +/- 21, 51 +/- 21, 51 +/- 23, 48 +/- 20, 56 +/- 24, and 48 +/- 20 microliters/min, at 90, 105, 120, 135, 150, 165, and 180 min, respectively. These values did not differ significantly from each other. In group II (bumetanide group), after baseline control CSF production had been determined at 90 and 105 min, bumetanide (10(-4) mol/liter) was added to VCP. Mean values for CSF production were 54 +/- 15 and 52 +/- 17 microliters/min before, and 39 +/- 25, 34 +/- 19, 28 +/- 10, 30 +/- 17, and 30 +/- 18 microliters/min after addition of bumetanide at 90, 105, 120, 135, 150, 165, and 180 min, respectively. Comparing the two groups, baseline values for CSF production measured at 90 and 105 min did not differ significantly. After addition of bumetanide (group II), however, decrements in CSF production varied from 30 +/- 27% at 120 min to 47 +/- 14% at 150 min, which were significantly different from changes in group I. The results of this study indicate that NaCl cotransport carrier is involved in secretion of CSF in dogs, and inhibition of the transporter results in approximately 50% reduction in CSF production. PMID- 8227342 TI - Charge selectivity of the glomerular filtration barrier in healthy and nephrotic humans. AB - We used dextran sulfate (DS) to evaluate barrier charge selectivity in 11 nonproteinuric subjects and in 11 patients with the nephrotic syndrome due to either membranous nephropathy or minimal change nephropathy. The 3H-DS preparation spanned a molecular radius interval of 10-24 A and exhibited size dependent protein binding in vitro. Urine and ultrafiltrates of plasma were separated by size into narrow fractions using gel permeation chromatography. The sieving coefficient (theta) for ultrafilterable DS of 15A radius averaged 0.68 +/ 0.03 in nonproteinuric vs. 0.95 +/- 0.05 in nephrotic subjects (P < 0.001). Uncharged dextrans of broad size distribution were used to evaluate barrier size selectivity in separate groups of nonproteinuric subjects (n = 19) and nephrotic patients with either minimal change (n = 20) or membranous nephropathy (n = 27). The value of theta for an uncharged dextran of similarly small radius (approximately 18 A) was significantly larger than that observed for DS in nonproteinuric subjects, but was similar in nephrotic individuals. Further, impaired barrier size-selectivity, as assessed by the sieving profile for uncharged dextrans (18-60 A radius), failed to account fully for the observed level of albuminuria in almost half of the patients with either minimal change (9/20) or membranous nephropathy (12/27). Together these findings suggest that the human glomerular capillary wall normally provides an electrostatic barrier to filtration of negatively charged macromolecules such as albumin, and that impairment of this electrostatic barrier contributes to the magnitude of albuminuria in the nephrotic syndrome. PMID- 8227343 TI - Metabolic effects of the nocturnal rise in cortisol on carbohydrate metabolism in normal humans. AB - Glucocorticoid concentrations vary throughout the day. To determine whether an increase in cortisol similar to that present during sleep is of physiologic significance in humans, we studied the disposition of a mixed meal when the nocturnal rise in cortisol was mimicked or prevented using metyrapone plus either a variable or constant hydrocortisone infusion. When glucose concentrations were matched with a glucose infusion, hepatic glucose release (2.6 +/- 0.2 vs. 1.5 +/- 0.4 nmol/kg per 6 h) was higher (P < 0.05) while glucose disappearance (5.9 +/- 0.3 vs. 7.3 +/- 0.9 mmol/kg per 6 h) and forearm arteriovenous glucose difference (64 +/- 24 vs. 231 +/- 62 mmol/dl per 6 h) were lower (P < 0.05) during the variable than basal infusion. The greater hepatic response during the variable cortisol infusion was mediated (at least in part) by inhibition of insulin and stimulation of glucagon secretion as reflected by lower (P < 0.05) C-peptide (0.29 +/- 0.01 vs. 0.38 +/- 0.04 mmol/liter per 6 h) and higher (P < 0.05) glucagon (42.7 +/- 2.0 vs. 39.3 +/- 1.8 ng/ml per 6 h) concentrations. In contrast, the decreased rates of glucose uptake appeared to result from a state of "physiologic" insulin resistance. The variable cortisol infusion also increased (P < 0.05) postprandial palmitate appearance as well as palmitate, beta hydroxybutyrate, and alanine concentrations, suggesting stimulation of lipolysis, ketogenesis, and proteolysis. We conclude that the circadian variation in cortisol concentration is of physiologic significance in normal humans. PMID- 8227344 TI - Novel splicing, missense, and deletion mutations in seven adenosine deaminase deficient patients with late/delayed onset of combined immunodeficiency disease. Contribution of genotype to phenotype. AB - We examined the genetic basis for adenosine deaminase (ADA) deficiency in seven patients with late/delayed onset of immunodeficiency, an underdiagnosed and relatively unstudied condition. Deoxyadenosine-mediated metabolic abnormalities were less severe than in the usual, early-onset disorder. Six patients were compound heterozygotes; 7 of 10 mutations found were novel, including one deletion (delta 1019-1020), three missense (Arg156 > His, Arg101 > Leu, Val177 > Met), and three splicing defects (IVS 5, 5'ss T+6 > A; IVS 10, 5'ss G+1 > A; IVS 10, 3'ss G-34 > A). Four of the mutations generated stop signals at codons 131, 321, 334, and 348; transcripts of all but the last, due to delta 1019-1020, were severely reduced. delta 1019-1020 (like delta 955-959, found in one patient and apparently recurrent) is at a short deletional hot spot. Arg156 > His, the product of which had detectable activity, was found in three patients whose second alleles were unlikely to yield active ADA. The oldest patient diagnosed was homozygous for a single base change in intron 10, which activates a cryptic splice acceptor, resulting in a protein with 100 extra amino acids. We speculate that this "macro ADA," as well as the Arg156 > His, Arg101 > Leu, Ser291 > Leu, and delta 1019-1020 products, may contribute to mild phenotype. Tissue-specific variation in splicing efficiency may also ameliorate disease severity in patients with splicing mutations. PMID- 8227345 TI - Cellular basis for the negative inotropic effects of tumor necrosis factor-alpha in the adult mammalian heart. AB - To define the mechanism(s) responsible for the negative inotropic effects of tumor necrosis factor-alpha (TNF alpha) in the adult heart, we examined the functional effects of TNF alpha in the intact left ventricle and the isolated adult cardiac myocyte. Studies in both the ventricle and the isolated adult cardiac myocyte showed that TNF alpha exerted a concentration- and time-dependent negative inotropic effect that was fully reversible upon removal of this cytokine. Further, treatment with a neutralizing anti-TNF alpha antibody prevented the negative inotropic effects of TNF alpha in isolated myocytes. A cellular basis for the above findings was provided by studies which showed that treatment with TNF alpha resulted in decreased levels of peak intracellular calcium during the systolic contraction sequence; moreover, these findings did not appear to be secondary to alterations in the electrophysiological properties of the cardiac myocyte. Further studies showed that increased levels of nitric oxide, de novo protein synthesis, and metabolites of the arachidonic acid pathway were unlikely to be responsible for the TNF alpha-induced abnormalities in contractile function. Thus, these studies constitute the initial demonstration that the negative inotropic effects of TNF alpha are the direct result of alterations in intracellular calcium homeostasis in the adult cardiac myocyte. PMID- 8227346 TI - Mononuclear cell infiltration and its relation to the expression of major histocompatibility complex antigens and adhesion molecules in pancreas biopsy specimens from newly diagnosed insulin-dependent diabetes mellitus patients. AB - We examined pancreas biopsy specimens from 18 newly diagnosed insulin-dependent diabetes mellitus (IDDM) patients to elucidate the mechanism underlying beta cell destruction. Pancreas islets were seen in all patients and insulitis in eight patients. Infiltrating mononuclear cells consisted of CD4+T, CD8+T, B lymphocytes, and macrophages. Among them, CD8+T lymphocytes were predominant and macrophages followed. The expression of MHC class I antigens was increased in islet and endothelial cells in nine patients. MHC class II expression was increased in endothelial cells of the same patients. The expression of intercellular adhesion molecule-1 was increased in endothelial cells in two of the nine patients with MHC hyperexpression; in one of them, lymphocyte function associated antigen-3 expression was also increased. Out of the eight patients with insulitis, seven showed MHC class I hyper-expression, whereas 2 of the 10 patients without insulitis showed the phenomenon (P < 0.05). The relation between insulitis and the hyperexpression of adhesion molecules was not evident. In conclusion, we revealed the close relation between CD8+T lymphocyte-predominant insulitis and MHC class I hyperexpression in islet cells. This suggests that infiltrating CD8+T lymphocytes recognize islet autoantigens in association with increased MHC class I molecules and act as major effector cells in autoimmune response against islet cells in IDDM pancreases. The role of adhesion molecules in the pathogenesis of IDDM still remains to be elucidated. PMID- 8227347 TI - Effects of interleukin-1 on syntheses of alkaline phosphatase, type X collagen, and 1,25-dihydroxyvitamin D3 receptor, and matrix calcification in rabbit chondrocyte cultures. AB - The effect of IL-1 on expression of the mineralization-related phenotype by chondrocytes was examined. In cultures of rabbit growth plate chondrocytes, IL-1 beta at 0.1 ng/ml caused 95% decreases in alkaline phosphatase activity, alkaline phosphatase mRNA levels, the incorporation of 45Ca into insoluble material, and the calcium content during the hypertrophic stage. These effects of IL-1 beta were dose-dependent and were observed in 24-48 h. Furthermore, IL-1 beta suppressed increase in cell size and the syntheses of 1,25-dihydroxyvitamin D3 receptor and type X collagen, other markers of hypertrophy, but had little effect on the synthesis of total protein including type II collagen. The inhibition of calcification was observed only when chondrocytes were exposed to IL-1 before the onset of calcification: IL-1 treatment from the mineralization stage had a marginal effect on 45Ca incorporation into insoluble material. These results suggest that IL-1 inhibits chondrocyte hypertrophy and the onset of calcification in ossifying cartilage. PMID- 8227348 TI - Antiidiotypic antibodies to sperm in sera of fertile women that neutralize antisperm antibodies. AB - The presence of antiidiotypic antibodies (ab-2) to sperm was investigated in the sera of fertile, infertile, and virgin women using sperm-specific anti-FA-1 monoclonal antibody Fab'.ab-2 were detected in 71% (17/24) of sera from fertile women and in none (0/12) of the sera from virgin females by the enzyme-linked immunosorbent assay, Western blot procedure, and immunoprecipitation procedure. Sera from infertile women that had antisperm antibodies showed a minimal presence of ab-2, with only three sera (13%, 3/23) demonstrating the presence of low levels of ab-2. The ab-2 present in fertile women were capable of neutralizing the fertilization-inhibitory activity of anti-FA-1 antibody in a concentration dependent manner in a human sperm penetration assay (SPA) of zona-free hamster oocytes. ab-2 were also capable of inhibiting the binding of antisperm antibodies to the sperm surface as determined by the immunobead binding technique. This is the first report demonstrating the presence of ab-2 in the sera of fertile women that are capable of neutralizing antisperm antibodies present in sera of infertile women. These findings suggest that the inability to detect antisperm antibody activity in the sera of fertile women may be due to higher levels of ab 2 present in these sera than levels found in sera of infertile women, although both groups may be producing antisperm antibody response after sexual exposure to sperm. PMID- 8227349 TI - Different localization and regulation of two types of vasopressin receptor messenger RNA in microdissected rat nephron segments using reverse transcription polymerase chain reaction. AB - Recent studies have revealed that arginine vasopressin (AVP) has at least two types of receptors in the kidney: V1a receptor and V2 receptor. In this study, microlocalization of mRNA coding for V1a and V2 receptors was carried out in the rat kidney using a reverse transcription and polymerase chain reaction. Large signals for V1a receptor PCR product were detected in the glomerulus, initial cortical collecting duct, cortical collecting duct, outer medullary collecting duct, inner medullary collecting duct, and arcuate artery. Small but detectable signals were found in proximal convoluted and straight tubules, inner medullary thin limbs, and medullary thick ascending limbs. Large signals for V2 receptor mRNA were detected in the cortical collecting duct, outer medullary collecting duct, and inner medullary collecting duct. Small signals for V2 receptor were found in the inner medullary thick limbs, medullary thick ascending limbs, and initial cortical collecting duct. Next, we investigated V1a and V2 receptor mRNA regulation in the dehydrated state. During a 72-h water restriction state, the plasma AVP level increased and V2 receptor mRNA decreased in collecting ducts. In contrast, V1a receptor mRNA did not change significantly. Thus, the two AVP receptor subtypes are distributed differently along the nephron, and these mRNAs are regulated differently in the dehydrated state. PMID- 8227350 TI - Interleukin-6 functions as an intracellular growth factor in hairy cell leukemia in vitro. AB - The role of interleukin-6 (IL-6) in the growth of B cell derived hairy cell leukemia (HCL) was characterized. Purified hairy cells (HCs) did not increase DNA synthesis in vitro in response to exogenous IL-6; however, they expressed IL-6 receptor (IL-6R) mRNA and bound directly fluorochrome labeled IL-6. IL-6 mRNA was not detectable in tumor cells by Northern blotting, but was evident using PCR amplification. Although intracytoplasmic IL-6 protein was not demonstrable, HCs did secrete low levels of IL-6. Neutralizing antibody to IL-6 did not inhibit HC DNA synthesis. Since tumor necrosis factor (TNF) is a growth factor for HCL, we determined whether the TNF effect could be IL-6-mediated. TNF markedly augmented in vitro DNA synthesis by HCs. TNF did not alter IL-6R expression or IL-6 binding; however, IL-6 mRNA and IL-6 protein were detectable after 3-d culture of HCs with TNF. In addition, IL-6 secretion by HCs was markedly augmented by TNF. Finally, although neither IL-6 nor anti-IL-6 antibody altered TNF-induced DNA synthesis by HCs, IL-6 antisense oligonucleotide inhibited TNF-induced DNA synthesis and IL-6 secretion by HCs. Therefore, IL-6 does not directly affect the growth of HCL, but rather mediates TNF-induced DNA synthesis via an intracytoplasmic mechanism. PMID- 8227351 TI - Limb congestion and sympathoexcitation during exercise. Implications for congestive heart failure. AB - During static exercise, heart failure (HF) subjects activate the sympathetic nervous system differently than normal controls. HF causes metaboreceptor desensitization with either enhanced mechanoreceptor activity or central command. In this report, we examined whether increased muscle interstitial pressure, as seen in HF, augments other neural systems. We measured muscle sympathetic nerve activity (MSNA; peroneal nerve) in 10 normals during static exercise (40% maximal voluntary grip) and posthandgrip circulatory arrest (PHG-CA). This was repeated after venous congestion (VC; cuff inflation to 90 mmHg). VC increased forearm volume (plethysmography) by 4.7%. MSNA responses to exercise were greater after VC (150.5 +/- 41.8 vs. 317.3 +/- 69.9 arbitrary units; P < 0.01). However, MSNA responses during PHG-CA were not affected by VC, and 31P nuclear magnetic resonance (n = 5) demonstrated no effect of VC on pH or H2PO4-. Similar effects of VC on MSNA were noted after ischemic exercise (n = 7), excluding flow alterations as the explantation. VC probably sensitized mechanically sensitive afferents since MSNA during involuntary biceps contractions increased after VC (n = 6), and skin sympathetic nerve responses during handgrip, an index of central command, were not increased by VC (n = 6). PMID- 8227352 TI - Degradation of biomaterials by phagocyte-derived oxidants. AB - Polymers used in implantable devices, although relatively unreactive, may degrade in vivo through unknown mechanisms. For example, polyetherurethane elastomers used as cardiac pacemaker lead insulation have developed surface defects after implantation. This phenomenon, termed "environmental stress cracking," requires intimate contact between polymer and host phagocytic cells, suggesting that phagocyte-generated oxidants might be involved. Indeed, brief exposure of polyetherurethane to activated human neutrophils, hypochlorous acid, or peroxynitrite produces modifications of the polymer similar to those found in vivo. Damage to the polymer appears to arise predominantly from oxidation of the urethane-aliphatic ester and aliphatic ether groups. There are substantial increases in the solid phase surface oxygen content of samples treated with hypochlorous acid, peroxynitrite or activated human neutrophils, resembling those observed in explanted polyetherurethane. Furthermore, both explanted and hypochlorous acid-treated polyetherurethane show marked reductions in polymer molecular weight. Interestingly, hypochlorous acid and peroxynitrite appear to attack polyetherurethane at different sites. Hypochlorous acid or activated neutrophils cause decreases in the urethane-aliphatic ester stretch peak relative to the aliphatic ether stretch peak (as determined by infrared spectroscopy) whereas peroxynitrite causes selective loss of the aliphatic ether. In vivo degradation may involve both hypohalous and nitric oxide-based oxidants because, after long-term implantation, both stretch peaks are diminished. These results suggest that in vivo destruction of implanted polyetherurethane involves attack by phagocyte-derived oxidants. PMID- 8227353 TI - In vivo stimulation of connective tissue accumulation by the tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+ in rat experimental wounds. AB - The tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+ (GHK-Cu) was first described as a growth factor for differentiated cells. Recent in vitro data showed that it possesses several properties of a potential activator of wound repair. We investigated the effects of GHK-Cu in vivo, using the wound chamber model described previously (Schilling, J.A., W. Joel, and M.T. Shurley, 1959. Surgery [St. Louis]. 46:702-710). Stainless steel wire mesh cylinders were implanted subcutaneously on the back of rats. The animals were divided into groups that received sequential injections into the wound chamber of either saline (control group) or various concentrations of GHK-Cu. At the end of the experiments, rats were killed, wound chambers were collected, and their content was analyzed for dry weight, total proteins, collagen, DNA, elastin, glycosaminoglycans, and specific mRNAs for collagens and TGF beta. In the GHK-Cu injected wound chambers, a concentration-dependent increase of dry weight, DNA, total protein, collagen, and glycosaminoglycan contents was found. The stimulation of collagen synthesis was twice that of noncollagen proteins. Type I and type III collagen mRNAs were increased but not TGF beta mRNAs. An increase of the relative amount of dermatan sulfate was also found. A control tripeptide, L glutamyl-L-histidyl-L-proline, had no significant effect. These results demonstrate that GHK-Cu is able to increase extracellular matrix accumulation in wounds in vivo. PMID- 8227354 TI - Two different cytochrome P450 enzymes are the adrenal antigens in autoimmune polyendocrine syndrome type I and Addison's disease. AB - Autoimmune polyendocrine syndrome type I (APS I) and idiopathic Addison's disease are both disorders with adrenal insufficiency but with differences in genetic background, clinical presentation, and extent of extraadrenal manifestations. In this study the major adrenal autoantigen identified with sera from patients with APS I was characterized by analyses using indirect immunofluorescence, Western blots of adrenal subcellular fractions and of recombinant proteins, immunoprecipitations of [35S]methionine-labeled lysates of a human steroid producing cell line, and studies of enzymatic activity. Sera from patients with APS I, identifying cells in adrenal glands and testes involved in steroid synthesis, reacted in Western blots with a 53-kD antigen, which comigrated with the cytochrome P450 cholesterol side chain cleavage enzyme (SCC). The sera also immunoprecipitated this protein from lysates of radiolabeled adrenal cells. The enzymatic activity of SCC was inhibited by the APS I sera but not by control sera. Sera from patients with idiopathic Addison's disease did not react with the SCC. The results show that the autoimmune responses towards adrenal tissue in patients suffering from APS I and Addison's disease are remarkably selective and suggest that a determination of the antigen involved in a patient with autoimmune adrenal insufficiency will have diagnostic as well as prognostic implications. PMID- 8227355 TI - Involvement of the tyrosinase gene in the deposition of cardiac lipofuscin in mice. Association with aortic fatty streak development. AB - Lipofuscin pigment, a terminal oxidation product, accumulates within cells during the normal aging process and under certain pathological conditions. We have analyzed a genetic cross between two inbred mouse strains, BALB/cJ and a subline of C57BL/6J, which differ in lipofuscin deposition. A comparison of the segregation pattern of cardiac lipofuscin with the albino locus (c) on mouse chromosome 7 revealed complete concordance. Analysis of spontaneous mutants of the tyrosinase gene, encoded by the albino locus, confirmed that the tyrosinase gene itself controls lipofuscin formation. Genetic analysis of other strains indicated that one or more additional genes cab contribute to the inheritance of lipofuscin. We also present evidence for an association between cardiac lipofuscin deposition and aortic fatty streak development in the mouse. PMID- 8227356 TI - Gene transfer into respiratory epithelial cells by targeting the polymeric immunoglobulin receptor. AB - A system for targeting foreign DNA to epithelial cells in vitro has been developed by exploiting receptor-mediated endocytosis. The polymeric immunoglobulin receptor transports dimeric immunoglobulin A and immunoglobulin M through epithelial cells, including those of the respiratory tract, by binding the immunoglobulins at the basolateral surface and transporting them across the cell. Fab fragments of antibodies directed against the extracellular portion of the receptor, secretory component, are similarly transported. Anti-human secretory component Fab fragments were covalently linked to a polycation, and complexed to various expression plasmids. When bound to an expression plasmid containing the Escherichia coli lacZ gene ligated to the Rous sarcoma virus promoter, the complexes transfected HT29.74 human colon carcinoma cells induced to express polymeric immunoglobulin receptor, but not those lacking the receptor. Primary cultures of human tracheal epithelial cells grown on collagen gels, which induce the expression of polymeric immunoglobulin receptor, were also transfected with the complexes. From 5 to 66% of the respiratory epithelial cells had beta galactosidase activity after treatment, comparable to the percentage of cultured human tracheal epithelial cells that express polymeric immunoglobulin receptor (8 35%). The addition of excess human secretory component (Fab ligand) to the culture medium at the time of transfection blocked the delivery of DNA. The expression plasmid, either alone, complexed to the polycation, or complexed to a carrier based on an irrelevant Fab fragment, was not effective in transfecting either cell type. This DNA carrier system introduces DNA specifically into epithelial cells that contain pIgR in vitro. PMID- 8227357 TI - Development of pemphigus vulgaris-like lesions in severe combined immunodeficiency disease mice reconstituted with lymphocytes from patients. AB - Pemphigus vulgaris is an autoimmune blistering disease that is induced by binding of antibodies to a 130/85-kD protein complex on epidermal keratinocytes. An in vivo experimental model of this disease was developed by reconstituting severe combined immunodeficiency (SCID) mice with 1-10 x 10(7) PBL from patients with naturally occurring pemphigus vulgaris. Of 49 reconstituted mice, 34 (69%) produced human IgG levels of > 0.1 mg/ml. Circulating anti-pemphigus antibodies were found in 20 of the 34 successfully reconstituted mice; 44% of these animals had deposits of human IgG in their own skin after it was traumatized by either heat or cold. Spontaneous pemphigus vulgaris-like blisters associated with human IgG deposits were rarely found in mouse skin. By contrast, allogeneic human skin grafted to 10 to 12 mice before reconstitution with patients' PBL developed pemphigus vulgaris-like lesions containing human IgG deposits. These results demonstrate that SCID mice can serve as a model of an antibody-mediated human autoimmune skin disease. PMID- 8227358 TI - Novel nuclear autoantigen with splicing factor motifs identified with antibody from hepatocellular carcinoma. AB - A patient with liver cirrhosis who progressed to hepatocellular carcinoma was found to develop novel antinuclear antibodies. The serum was used to isolate full length cDNA clones encoding related proteins of 530 amino acids (representative clone HCC1.4) and 524 amino acids (representative clone HCC1.3). Affinity purified antibodies eluted from recombinant proteins recognized a 64-kD nuclear protein in Western blotting and decorated the nucleoplasm in a speckled-network fashion in immunofluorescence, colocalizing with antibodies to pre-mRNA splicing factor SC35 and uridine-rich small nuclear RNAs. The deduced amino acid sequence contained an arginine/serine-rich (RS) domain and three-ribonucleoprotein consensus sequence domains, two classes of motifs present in several splicing factors. A repeating octapeptide of Arg-Ser-Arg-Ser-Arg(Lys)-Glu(Asp)-Arg Lys(Arg) was present in RS region of HCC1. This octapeptide sequence called RS ERK motif was also found in splicing factors U2AF 35- and 65-kD proteins and 70 kD U1 small nuclear ribonucleoprotein. The molecular features and immunolocalization data suggest that the HCC1 autoantigen may be associated with splicing activities and are consistent with observations that autoantibody responses frequently target molecules involved in important cellular biosynthetic functions. PMID- 8227359 TI - Alkylating agents and immunotoxins exert synergistic cytotoxic activity against ovarian cancer cells. Mechanism of action. AB - Alkylating agents can be administered in high dosage to patients with ovarian cancer using autologous bone marrow support, but drug-resistant tumor cells can still persist. Immunotoxins provide reagents that might eliminate drug resistant cells. In the present study, concurrent treatment with alkylators and immunotoxins proved superior to treatment with each agent alone. Toxin immunoconjugates prepared from different monoclonal antibodies and recombinant ricin A chain (rRTA) inhibited clonogenic growth of ovarian cancer cell lines in limiting dilution assays. When alkylating agents and toxin conjugates were used in combination, the addition of the immunotoxins to cisplatin, or to cisplatin and thiotepa, produced synergistic cytotoxic activity against the OVCA 432 and OVCAR III cell lines. Studies performed to clarify the mechanism of action showed that cisplatin and thiotepa had no influence on internalization and binding of the 317G5-rRTA immunotoxin. Intracellular uptake of [195m]Pt-cisplatin was not affected by the immunoconjugate and thiotepa. The combination of the 317G5-rRTA and thiotepa, as well as 317G5-rRTA alone, increased [195m]Pt cisplatin-DNA adduct levels. The immunotoxin alone and in combination with the alkylators decreased intracellular glutathione levels and reduced glutathione-S-transferase activity. Repair of DNA damage induced by the combination of alkylators and 317G5 rRTA was significantly reduced when compared to repair after damage with alkylators alone. These findings suggest that immunotoxins affect levels and activity of enzymes required for the prevention and repair of alkylator damage. PMID- 8227360 TI - Low protein diet alters urea transport and cell structure in rat initial inner medullary collecting duct. AB - Low protein diets reverse the urea concentration gradient in the renal inner medulla. To investigate the mechanism(s) for this change, we studied urea transport and cell ultrastructure in initial and terminal inner medullary collecting ducts (IMCD) from rats fed 18% protein or an isocaloric, 8% protein diet for 4 wk. Serum urea, aldosterone, and albumin were significantly lower in rats fed 8% protein, but total protein and potassium were unchanged. Vasopressin stimulated passive urea permeability (Purea) threefold (P < 0.05) in initial IMCDs from rats fed 8% protein, but not from rats fed 18% protein. Luminal phloretin reversibly inhibited vasopressin-stimulated Purea. However, in terminal IMCDs from rats fed either diet, vasopressin stimulated Purea. Net transepithelial urea flux (measured with identical perfusate and bath solutions) was found only in initial IMCDs from rats fed 8% protein. Reducing the temperature reversibly inhibited it, but phloretin did not. Electron microscopy of initial IMCD principal cells from rats fed 8% protein showed expanded Golgi bodies and prominent autophagic vacuoles, and morphometric analysis demonstrated a marked increase in the surface density and boundary length of the basolateral plasma membrane. These ultrastructural changes were not observed in the terminal IMCD. Thus, 8% dietary protein causes two new urea transport processes to appear in initial but not terminal IMCDs. This is the first demonstration that "active" urea transport can be induced in a mammalian collecting duct segment. PMID- 8227361 TI - Hereditary isolated glucocorticoid deficiency is associated with abnormalities of the adrenocorticotropin receptor gene. AB - Isolated glucocorticoid deficiency (IGD) is an autosomal recessive disorder characterized by progressive primary adrenal insufficiency, without mineralocorticoid deficiency. The cDNA and gene of the human ACTH receptor were recently cloned. The gene encodes a 297-amino acid protein that belongs to the G protein-coupled superfamily of membrane receptors. We hypothesized that the ACTH receptor gene might be defective in IGD. To examine this, we studied its genomic structure by PCR and direct sequencing in a 5-yr-old proband with the disease, his parents, and grandparents. The proband was a compound heterozygote for two different point mutations, one in each allele: (a) a substitution (C-->T), also found in one allele of the mother and maternal grandmother, which introduced a premature stop codon (TGA) at position 201 of the protein; this mutant receptor lacks its entire carboxy-terminal third and, if expressed, should be unable to transduce the signal; and (b) a substitution (C-->G), also found in one of the paternal alleles, which changed neutral serine120 in the apolar third transmembrane domain of the receptor to a positively charged arginine, probably disrupting the ligand-binding site. Standard ovine corticotropin releasing hormone (oCRH) test in the heterozygote parents and maternal grandmother revealed exaggerated and prolonged ACTH responses, suggestive of subclinical resistance to ACTH. We conclude that IGD in this family appears to be due to defects of the ACTH receptor gene. The oCRH test appears to be useful in ascertaining heterozygosity in this syndrome. PMID- 8227362 TI - Thromboxane-insensitive dog platelets have impaired activation of phospholipase C due to receptor-linked G protein dysfunction. AB - Human platelet thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptors are linked to phosphoinositide-specific phospholipase C (PI-PLC) via a G protein tentatively identified as a member of the Gq class. In contrast, platelet thrombin receptors appear to activate PI-PLC via other unidentified G proteins. Platelets from most dogs are TXA2 insensitive (TXA2-); i.e., they do not aggregate irreversibly or secrete although they bind TXA2, but they respond normally to thrombin. In contrast, a minority of dogs have TXA2-sensitive (TXA2+) platelets that are responsive to TXA2. To determine the mechanism responsible for TXA2- platelets, we evaluated receptor activation of PI-PLC. Equilibrium binding of TXA2/PGH2 receptor agonists, [125I]BOP and [3H]U46619, and antagonist, [3H]SQ29,548, revealed comparable high-affinity binding to TXA2-, TXA2+, and human platelets. U46619-induced PI-PLC activation was impaired in TXA2- platelets as evidenced by reduced (a) phosphorylation of the 47-kD substrate of protein kinase C, (b) phosphatidic acid (PA) formation, (c) rise in cytosolic calcium concentration, and (d) inositol 1,4,5 trisphosphate (IP3) formation, while thrombin-induced PI PLC activation was not impaired. GTPase activity stimulated by U46619, but not by thrombin, was markedly reduced in TXA2- platelets. Antisera to Gq class alpha subunits abolished U46619-induced GTPase activity in TXA2-, TXA2+, and human platelets. Direct G protein stimulation by GTP gamma S yielded significantly less PA and IP3 in TXA2- platelets. Immunotransfer blotting revealed comparable quantities of Gq class alpha-subunits in all three platelet types. Thus, TXA2- dog platelets have impaired PI-PLC activation in response to TXA2/PGH2 receptor agonists secondary to G protein dysfunction, presumably involving a member of the Gq class. PMID- 8227363 TI - Effects of deoxygenation on active and passive Ca2+ transport and on the cytoplasmic Ca2+ levels of sickle cell anemia red cells. AB - Elevated [Ca2+]i in deoxygenated sickle cell anemia (SS) red cells (RBCs) could trigger a major dehydration pathway via the Ca(2+)-sensitive K+ channel. But apart from an increase in calcium permeability, the effects of deoxygenation on the Ca2+ metabolism of sickle cells have not been previously documented. With the application of 45Ca(2+)-tracer flux methods and the combined use of the ionophore A23187, Co2+ ions, and intracellular incorporation of the Ca2+ chelator benz-2, in density-fractionated SS RBCs, we show here for the first time that upon deoxygenation, the mean [Ca2+]i level of SS discocytes was significantly increased, two- to threefold, from a normal range of 9.4 to 11.4 nM in the oxygenated cells, to a range of 21.8 to 31.7 nM in the deoxygenated cells, closer to K+ channel activatory levels. Unlike normal RBCs, deoxygenated SS RBCs showed a two- to fourfold increase in pump-leak Ca2+ turnover. Deoxygenation of the SS RBCs reduced their Ca2+ pump Vmax, more so in reticulocyte- and discocyte-rich than in dense cell fractions, and decreased their cytoplasmic Ca2+ buffering. Analysis of these results suggests that both increased Ca2+ influx and reduced Ca2+ pump extrusion contribute to the [Ca2+]i elevation. PMID- 8227364 TI - Cytokine-stimulated secretion of group II phospholipase A2 by rat mesangial cells. Its contribution to arachidonic acid release and prostaglandin synthesis by cultured rat glomerular cells. AB - Potent pro-inflammatory cytokines, such as interleukin 1 (IL-1) or tumor necrosis factor (TNF) alpha have been found to increase group II phospholipase A2 (PLA2) synthesis and secretion by mesangial cells. In all cases 85-90% of the enzyme is secreted from the cells and a parallel increase in prostaglandin (PG)E2 synthesis is observed. We report here that co-incubation with a monoclonal antibody that specifically binds and neutralizes rat group II PLA2 attenuates IL-1 beta and TNF alpha-stimulated PGE2 production by 45% and 52%, respectively. CGP43182, a specific inhibitor of group II PLA2, potently blocks mesangial cell group II PLA2 in vitro with a half-maximal inhibitory concentration (IC50) of 1.5 microM, while only slightly affecting mesangial cell high molecular weight PLA2. CGP 43182 markedly attenuates IL-1 beta- and TNF alpha-stimulated PGE2 synthesis in intact mesangial cells with IC50's of 1.3 and 1.0 microM, respectively. PLA2 secreted from cytokine-stimulated mesangial cells was purified to homogeneity. Addition of the purified enzyme to unstimulated mesangial cells causes a marked release of arachidonic acid and a subsequent increased synthesis of PGE2. Moreover, addition of purified PLA2 to a cloned rat glomerular epithelial cell line and cultured bovine glomerular endothelial cells augmented both arachidonic acid release and PGE2 synthesis, with the endothelial cells being especially sensitive. Thus, cytokine-triggered synthesis and secretion of group II PLA2 by mesangial cells contributes, at least in part, to the observed synthesis of PGE2 that occurs in parallel to the enzyme secretion. Furthermore, extracellular PLA2 secreted by mesangial cells is able to stimulate arachidonic acid release and PGE2 synthesis by the adjacent endothelial and epithelial cells. These data suggest that expression and secretion of group II PLA2 triggered by pro-inflammatory cytokines may crucially participate in the pathogenesis of inflammatory processes within the glomerulus. PMID- 8227365 TI - Cell-specific regulation of type II phospholipase A2 expression in rat mesangial cells. AB - IL-1 stimulates mesangial cells to synthesize specific proteins, including a non pancreatic (Type II) phospholipase A2 (PLA2). We have studied the regulation of PLA2 by proinflammatory mediators, implicated in the pathogenesis of glomerulonephritis, and have assessed whether the activation of second messenger systems modulates or mimics PLA2 gene expression by cytokines. IL-1 alpha and beta, TNF alpha, and LPS, but not serum, IL-2, or PDGF, potently induce PLA2 mRNA, and enzyme expression. IL-1-stimulated mesangial cells express a 1.0 kB PLA2 mRNA transcript that is induced in a dose- and time-dependent manner. IL-1 stimulated increases in steady-state PLA2 mRNA abundance result from a moderate increase in PLA2 transcription rate that is amplified by the prolonged persistence of the transcript. Forskolin and dibutyryl cAMP potentiate IL-1 induced PLA2 mRNA and enzyme expression, but have no effect in the absence of cytokine. 12-tetradecanoyl phorbol 13-acetate, sn-1, 2-dioctanoyl glycerol or 1 oleoyl-2-acetyl-sn-glycerol fail to induce PLA2 expression or to alter the effect of IL-1 when coincubated with the cytokine. In contrast, serum deprivation for 24 h specifically enhances IL-1-stimulated PLA2. Genistein potentiates PLA2 mRNA expression in cells exposed to both IL-1 and serum. The inhibitory effect of serum on IL-1-induced PLA2 mRNA abundance is reproduced by PDGF but not dexamethasone. These data demonstrate that the signaling pathways directly engaged by IL-1 to induce PLA2 expression in mesangial cells interact with several second messenger systems in a cell-specific manner. We speculate that IL 1 induces specialized changes in mesangial cell structure and function through direct activation of a transcription factor(s), that result in induction of a specific gene set. PMID- 8227366 TI - Binding of cytokines to pharmaceutically prepared human immunoglobulin. AB - Pharmaceutically prepared IgG, pooled from sera of over 2,000 normal individuals, contained both monomeric and dimeric IgG. Each type of IgG bound 125I-labeled interleukin (IL)-1 alpha, IL-1 beta, IL-6, and tumor necrosis factor (TNF)-alpha. Increased binding to IgG was observed if 125I-IL-1 beta was denatured by heating to 39 degrees C. However, the binding of both nondenatured and denatured 125I-IL 1 beta was not inhibited by unlabeled IL-1 beta. In contrast, binding of 125I-IL 1 alpha, 125I-IL-6, and 125I-TNF alpha was inhibited by the corresponding unlabeled cytokine. Papain-digestion of IgG abolished binding of 125I-TNF alpha but failed to influence the displaceable binding of 125I-IL-1 alpha and 125I-IL 6. 125I-TNF alpha was a mixture of trimeric and monomeric forms, the latter being the predominant form at lower concentrations. The apparent saturability of 125I TNF alpha was explained by a higher nonspecific binding of monomeric than of trimeric 125I-TNF alpha to IgG. The amounts of cytokine antibodies in IgG preparations would contribute approximately 2 micrograms anti-IL-1 alpha IgG and 1 microgram anti-IL-6 IgG per kg body wt during high dose immune globulin therapy. In conclusion, pharmaceutical preparations of human IgG contain specific and neutralizing, high affinity antibodies against IL-1 alpha and IL-6, but not against TNF alpha or IL-1 beta. There are significant methodological pitfalls that hamper detection of IgG autoantibodies against cytokines. PMID- 8227367 TI - Protease nexin-2/amyloid beta protein precursor. A tight-binding inhibitor of coagulation factor IXa. AB - Protease nexin-2/amyloid beta protein precursor (PN-2/A beta PP) is an abundant, secreted platelet protein which is a potent inhibitor of coagulation Factor XIa. We examined other potential anticoagulant activities of PN-2/A beta PP. Purified Kunitz protease inhibitor domain of PN-2/A beta PP and PN-2/A beta PP itself were found to prolong the coagulation time of plasma and pure Factor IXa. The Kunitz protease inhibitor domain also inhibited the ability of Factor IXa to activate Factor X. PN-2/A beta PP inhibited Factor IXa with a Ki of 7.9 to 3.9 x 10(-11) M in the absence and presence of heparin, respectively. When the second-order rate constant of PN-2/A beta PP's inhibition of Factor IXa (2.7 x 10(8) M-1min-1) was compared to that of antithrombin III (3.8 x 10(6) M-1min-1), PN-2/A beta PP was at least a 71-fold more potent inhibitor of Factor IXa than antithrombin III. PN 2/A beta PP formed a complex with Factor IXa as detected by gel filtration and ELISA. The finding that PN-2/A beta PP is a potent inhibitor of Factor IXa could help to explain the spontaneous intracerebral hemorrhages seen in patients with hereditary cerebral hemorrhage with amyloidosis Dutch-type where there is an extensive accumulation of PN-2/A beta PP in their cerebral blood vessels. PMID- 8227368 TI - Endotoxin increases parathyroid hormone-related protein mRNA levels in mouse spleen. Mediation by tumor necrosis factor. AB - Parathyroid hormone-related protein (PTHrP) causes hypercalcemia in malignancy. However, the role and regulation of PTHrP in normal physiology is just beginning to be explored. PTHrP is found in the spleen and has several other features common to cytokines. Since endotoxin (LPS) causes many of its effects indirectly by inducing cytokines, studies were undertaken to determine whether LPS might also induce splenic PTHrP expression. LPS (100 ng/mouse) increased splenic PTHrP mRNA levels 3.6-fold in C3H/OuJ mice. This effect was maximal at 2 h and returned to baseline by 4 h. PTHrP peptide levels also increased 3.3-fold in splenic extracts in response to LPS (1 microgram/mouse). Murine TNF-alpha and human IL-1 beta, cytokines that mediate many of the effects of LPS, also increased splenic PTHrP mRNA levels. LPS-resistant C3H/HeJ mice, which produce minimal amounts of TNF and IL-1 in response to LPS, were resistant to LPS induction of splenic PTHrP mRNA, while TNF-alpha and IL-1 beta readily increased PTHrP mRNA levels in C3H/HeJ mice. Anti-TNF antibody blocked LPS induction of splenic PTHrP mRNA in C3H/OuJ mice by 68%, indicating that TNF is a mediator of the LPS induction of PTHrP levels. In contrast, an IL-1 receptor antagonist (IL-1ra) was ineffective. The increase in PTHrP in the spleen during the immune response suggests that PTHrP may play an important role in immune modulation, perhaps by mediating changes in lymphocyte proliferation and/or function. PMID- 8227369 TI - Hypertensive sodium-proton exchanger phenotype persists in immortalized lymphoblasts from essential hypertensive patients. A cell culture model for human hypertension. AB - An enhancement of sodium-proton exchange activity is a frequently observed ion transport abnormality in essential hypertension. The cellular basis for this has not yet been elucidated. Due to the lack of a specific cell culture system it has been impossible to distinguish between intrinsic cellular abnormalities and influences exerted by the hypertensive neurohumoral milieu. Using Epstein-Barr virus we have immortalized lymphocytes from controls and from patients with essential hypertension that exhibited enhanced sodium-proton exchanger activity. Sodium-proton exchanger activity was determined in cells loaded with the fluorescent cytosolic pH indicator 2'7'-biscarboxyethyl-5,6-carboxyfluorescein acetoxymethylester (BCECF) after pretreatment with 250 nM of the phorbol ester 12 O-tetradecanoylphorbol-13-acetate for 10 min. Cell lines from hypertensive patients displayed higher Vmax values of sodium-proton exchange than those from normotensive controls (129.6 +/- 30.0 vs. 77.1 +/- 13.2 mmol H+/min.; P < 0.001). Hill coefficients for H+ were distinctly lower in hypertension compared to normotension (1.12 +/- 0.12 vs. 1.50 +/- 0.14; P < 0.0001). The enhanced antiporter activity in cell lines from hypertensive patients was not accompanied by a corresponding increase in steady-state NHE-1 mRNA transcript levels, which argues against overexpression of antiporter protein in hypertension. The cells from hypertensive patients with high sodium-proton exchange activity proliferated distinctly faster than those from normotensive controls. These human cell lines represent a novel model to study the mutual interaction between sodium-proton exchange and cell proliferation, and may provide insights into the alterations in ion transport observed in a group of patients with essential hypertension. PMID- 8227370 TI - Reshaping of the skull in the early surgical correction of scaphocephaly. AB - It is possible to modify the mathematical patterns of projection geometry to reshape the skull in cranio-facial surgery. The technique of the 'gore-pattern' in accordance with the Hendel procedure changes the shape of individual sections of the skull by changing their radius of curvature. This technique was well known to engineers who built complex structures such as ships, hulls, airfoils and domes. A rigid form of the desired shape is the result to be achieved, which becomes permanent with healing. The gore-pattern is developed by cutting the surface of the sphere along longitudinal lines. Straightening each piece results in a series of elliptical segments referred to as 'gores'. When constructing a sphere from a flat sheet of material, each segment (gore) is bent through 180 degrees and then assembled side-by-side into a sphere. If the gore is cut in the middle, a hemisphere results. Because there are only small areas of contact between the gores, less bending force is necessary to fold a gore-pattern into a sphere than a zone-pattern. The interesting points of this technique, its advantages, and clinical application in the treatment of skull deformity in scaphocephaly are illustrated. PMID- 8227371 TI - Blunt trauma lesions of the extracranial internal carotid artery in patients with head injury. AB - Blunt trauma lesions of the extracranial internal carotid artery (ICA) are rare. In our hospital 18 patients with such an injury were diagnosed. All patients were involved in traffic accidents. Most of them had sustained head injuries with fractures of the skull, mandible or maxilla. The onset of neurological signs, most frequently hemiparesis, was usually delayed. 50 percent had bilateral ICA lesions but the clinical presentation was similar to those with unilateral lesions. Mortality of patients with ICA lesions was substantial (28%). PMID- 8227372 TI - Reconstruction of residual alveolo-palatal bone defects in cleft patients. A retrospective study. AB - The results of reconstruction of residual alveolo-palatal bone defects in 40 patients are evaluated clinically and radiographically in a retrospective study. Age at the time of surgery varied from 7.4 to 32.7 years. The patients were divided into two age groups: before and after 12 at the time of operation. Free autogenous anterior iliac crest bone grafts were used in 26 patients and free autogenous mandibular symphyseal bone grafts were used in 14 patients. The results show unsuccessful closure of the oronasal communication in the alveolo palatal area of the cleft in 3 out of 31 patients. In 3 out of 40 patients, radiographic incorporation of the bone graft was unsuccessful. The radiographic success was not related to increasing experience of the surgeons. The study indicates that the best grafting results are achieved when grafting is performed before 12 years of age. The radiographic results of mandibular symphyseal bone grafts tend to be slightly better compared with results obtained with anterior iliac crest bone. PMID- 8227373 TI - Treatment of mandibular fractures in the 1980s. AB - Treatment of 317 mandibular fracture patients between 1981 and 1990 at the Department of Oral and Maxillofacial Surgery, Institute of Dentistry, University of Oulu and Oulu University Hospital were analysed on the basis of patient files and radiographs. The patients had suffered altogether 452 mandibular fractures, most in the condylar region. The treatment was operative in 24% of cases and conservative in 76%. Operative treatment was commoner in edentulous than in partially edentulous or dentate patients. The proportion of operatively treated mandibular fractures increased toward the end of the period. The type of osteosynthesis also changed over the decade as wire and plate osteosynthesis gave way to compression plating and then to titanium miniplate and screw fixation of fractures. The advantages and disadvantages of the treatment methods are discussed and some typical cases using various modern fixation techniques are presented. PMID- 8227374 TI - Evaluation of mandibular fracture treatment. AB - A total of 124 patients treated for mandibular fractures were analyzed. Patients were divided into three groups according to treatment: intermaxillary fixation, wire fixation and mini-plate fixation. For each method, the success of treatment was evaluated with respect to surgical approach, fracture site and injury to treatment interval. Five basic parameters were used for evaluation of the outcome: occlusion, appearance, mastication, duration of IMF and complications. The treatment was surveyed based on both the surgeon's and patient's-evaluation. All parameters were scored and average values for every parameter calculated. The most successful treatment was achieved with mini-plate fixation in symphyseal and angle fractures. Intermaxillary fixation is indicated for mandibular body fractures with or without minimal displacement and a sufficient number of teeth. However, mini-plate fixation should be used for fractures with displacement. Wire fixation has been shown to be the poorest choice for all sites. Results showed that the intraoral approach has advantages over the extraoral one. The optimal time for treatment of mandibular fractures is within 72 h from time of injury. Even in fractures older than 7 days we recommended mini-plate fixation. PMID- 8227375 TI - The behaviour of neurogenic tumours of the maxillofacial region. AB - In 37 patients with neurogenic tumours of the maxillofacial region, epidemiology, clinical, sonographical and computer tomographical findings as well as the results of fine needle aspiration cytology (FNAC), histopathological evaluation of biopsies and flow cytometric determination of DNA ploidy, were analyzed in order to detect characteristics distinguishing the behaviour of the neoplasm prior to resection. Simultaneous histopathological and flow cytometric evaluation of biopsies is recommended to avoid the danger of mistaking ancient schwannomas for malignancies and vice versa. FNAC proved to be unreliable in these cases. The retromaxillary/pterygomandibular region appeared to be the most likely site for malignant schwannoma. Von Recklinghausen's neurofibromatosis in the patient or in their family should lead to immediate investigation of any mass developing in the vicinity of the facial or trigeminal nerves. PMID- 8227376 TI - Malignant fibrous histiocytoma of the head and neck. AB - Malignant fibrous histiocytomas are now recognized as the most common soft tissue sarcomas in adults. These tumors involve the head and neck region in 1 to 3% of cases. In this report we describe our experience with two typical and one unusual case of malignant fibrous histiocytoma of the head and neck region. In addition we review the diagnostic and therapeutic aspects of these tumors. PMID- 8227377 TI - Photoelastic analysis of internal screw fixation after sagittal split ramus osteotomy. AB - An experimental study on mechanical behavior and stability of internal screw fixation (ISF) of sagittal split ramus osteotomy has been carried out. A stress freezing method in a three-dimensional photoelastic experiment was used. Four ISF's that utilized different screw placement patterns were used for photoelastic analysis: (1) 3 screw linear pattern placed at 90 degrees to the lateral surface of the model; (2) triangular pattern at 90 degrees to the surface of the model, 2 above and 1 below the mandibular canal; (3) 2 screw linear pattern at 90 degrees, and (4) 3 screw oblique linear pattern at 75, 60 and 45 degrees to the surface of the model. The stress distribution differs markedly according to these 4 methods. PMID- 8227378 TI - High-resolution ultrasonography in the diagnosis of scrotal pathology: I. Normal scrotum and benign disease. AB - Ultrasonography is a readily accessible imaging modality. It is relatively inexpensive, is noninvasive, and requires no ionizing radiation or previous preparation. For these reasons, ultrasonography is the examination of choice in the study of the scrotum and its contents. We present a review of the pathology of the scrotum and its contents. For that, we cite multiple sources of the current medical literature and illustrate them with cases performed at the University of California Davis Medical Center during the last 2.5 years. Due to the extent of the pathology in consideration, we have divided it in two parts. This article deals with normal ultrasonography anatomy, technique, epididymal and testicular cysts, scrotal calcifications, acute and chronic inflammatory disease, testicular torsion, scrotal trauma, varicocele, undescended testis, and miscellaneous conditions. PMID- 8227379 TI - High-resolution ultrasonography in the diagnosis of scrotal pathology: II. Tumors. AB - Ultrasonography is a readily accessible imaging modality. It is relatively inexpensive, is noninvasive, and requires no ionizing radiation or previous preparation. For these reasons, ultrasound is the examination of choice in the study of the scrotum and its contents. We present a review of the pathology of the scrotum and its contents. For that, we cite multiple sources of the current medical literature and illustrate them with cases performed at the University of California Davis Medical Center During the last 2.5 years. This article deals with testicular and extratesticular tumors. Within testicular tumors, we describe primary tumors, such as those of germ cell and stromal origin, and secondary tumors of the lymphoproliferative and nonlymphoproliferative (metastatic) types. The most common extratesticular tumors are discussed. PMID- 8227380 TI - Correlation of antenatal ultrasonography and pathological examinations in 153 malformed fetuses. AB - Findings in 153 abnormal fetuses diagnosed by antenatal ultrasound and subsequently examined by a pediatric pathologist were reviewed. The primary diagnoses in the 153 cases were 67 nervous system anomalies, 19 gastrointestinal system anomalies, 18 genitourinary system anomalies, 13 chromosomal abnormalities, 10 skeletal system anomalies, and 26 other abnormalities. Forty six percent of the total number had multiple abnormalities. In 67 cases (44%), autopsy showed further anomalies or altered the ultrasonographic diagnosis. However, in only 36 (25%) were there autopsy findings that altered management. Errors in ultrasound diagnosis were most commonly associated with the presence of multiple anomalies and oligohydramnios. Expert pathological examination remains an essential tool in the management of fetal anomaly. PMID- 8227381 TI - Sigmoid diverticulitis in the female: transvaginal sonographic findings. PMID- 8227382 TI - Ultrasound and color flow Doppler manifestations of pseudoxanthoma elasticum. PMID- 8227383 TI - Preoperative sonographic diagnosis of gallbladder duplication: importance of challenge with fatty meal. PMID- 8227384 TI - Intra-amniotic membranes following amniocentesis. PMID- 8227385 TI - Uterine malignancy predicted by transvaginal sonography and color flow Doppler ultrasonography. PMID- 8227386 TI - Introduction to symposium on intravascular ultrasound assessment of the coronary arteries. PMID- 8227387 TI - Application of intracoronary ultrasonography in the study of coronary artery pathophysiology. AB - Coronary endothelial function, arterial distensibility, and elastic recoil were assessed in various groups of patients using intracoronary ultrasonic (ICUS) imaging. We found evidence of endothelial dysfunction in patients with risk factors for coronary artery disease even before atherosclerosis could be detected angiographically or with sensitive ICUS imaging. There was marked reduction of arterial distensibility in atherosclerotic arterial segments even with minor disease. Distensibility was partially restored in lesions following coronary balloon angioplasty (PTCA) or directional atherectomy (DCA). We also noted significantly greater elastic recoil following PTCA in soft lesions compared with hard lesions. We conclude that the use of ICUS imaging in cardiac catheterization laboratory provides useful information regarding various physiologic parameters. This capability will add new dimensions to the evaluation and management of patients with coronary artery disease. PMID- 8227388 TI - Mechanisms and outcomes of angioplasty and atherectomy assessed by intravascular ultrasound imaging. AB - Catheter-based intravascular ultrasound is a relatively new imaging tool to examine endovascular structure. One major goal for the development of intravascular ultrasound imaging has been to help clarify the mechanism of interventional therapies such as balloon angioplasty and directional atherectomy. Pathologic studies have suggested that plaque distribution and composition are key features that relate to initial and long-term success of coronary interventions. However, relatively little is known by angiography about the nature of plaque in the clinical setting. Intravascular ultrasound imaging provides a high resolution, "on-line" method of tracking the effects of catheter interventions such as balloon angioplasty and atherectomy. Because of its ability to visualize tissue beneath the luminal surface, ultrasound is generating new insights into the effect of plaque composition and distribution on the response to catheter therapies. With pre-procedure ultrasound imaging, it is increasingly possible to predict the result of a particular intervention, offering the potential for developing strategies of lesion-targeted therapy based on certain plaque characteristics. This article presents the early results of the GUIDE trial as a "works in progress" view of the role of intravascular ultrasound in helping interventionalists to understand--and to optimize--angioplasty and atherectomy. PMID- 8227389 TI - Lessons from intravascular ultrasonography: observations during interventional angioplasty procedures. AB - This article reviews many of the applications of intravascular ultrasonic imaging for coronary and peripheral arterial disease. In vitro studies demonstrate an excellent correlation between ultrasound measurements of lumen and plaque cross sectional area compared with histologic sections. In vivo clinical studies reveal the enhanced diagnostic capabilities of this technology compared with angiography. Ultrasonic imaging also permits visualization of the atherosclerotic plaque itself for the first time in vivo. In addition to accurately describing the plaque morphology, ultrasonography can identify some of the tissue characteristics of the plaque. During interventional procedures, ultrasonic imaging has been shown to be beneficial for enhanced diagnosis as well as improvement of our understanding of the mechanism of newer interventional devices such as directed atherectomy, rotational or TEC atherectomy, or excimer laser. Initial studies suggest that ultrasound guidance of intravascular stent deployment may be critical for optimizing stent placement. Randomized studies are currently in progress to determine whether the guidance provided by intravascular ultrasonic imaging will alter the results of interventional procedures so that the restenosis rate can be improved. PMID- 8227390 TI - Three-dimensional intravascular ultrasonography: reconstruction of endovascular stents in vitro and in vivo. AB - BACKGROUND: Intravascular ultrasonography displays an artery in loosely related cross-sectional images with limited axial information. However, intravascular ultrasound images are suited to three-dimensional reconstruction. METHODS: A comprehensive intravascular ultrasound imaging system was used to reconstruct planar images in three-dimensions. This system consisted of a 25MHz transducer tipped rigid probe (for in vitro studies) or a 25MHz transducer-tipped catheter within a 3.9F monorail imaging sheath (for in vivo studies), a motorized catheter pullback device that withdraws the transducer at 0.5mm/s, and an image processing computer that stacks 15 cross-sectional images/mm of stent axial length and then performs thresholding-based three-dimensional image rendering. We imaged 10 stents (4 Palmaz-Schatz, 3 Wiktor, 2 Strecker, and 1 Medinvent) in vitro after implantation in freshly harvested saphenous veins and 37 Palmaz-Schatz stents in vivo, 10 in native coronary arteries and 27 in vein grafts, 21 acutely and 18 on follow-up. RESULTS: Three-dimensional reconstruction of images obtained with this system reproduced the geometry of each stent design. In vitro, images of the Palmaz-Schatz stents showed the expanded diamonds, the central articulation, and flaring of both ends of both halves of the stents. Images of the Wiktor stents showed the sinusoidal wave-shaped coils in their helical configuration. Images of the Strecker stents showed the interlocking-loop design with gaps between the terminal loops at either end of the stent. Images of the Medivent stent reproduced the woven texture formed by braiding the stent wires. Three dimensional reconstruction of images obtained in vivo also reproduced the spatial geometry of the Palmaz-Schatz stent. However, reconstruction of in vivo images was limited by cardiac-cycle-linked vessel motion and torsion and the presence of echo-dense tissue that could not be separated completely from the stent itself. CONCLUSIONS: Properly acquired intravascular ultrasound images can be used to reconstruct the spatial geometry of endovascular stents. Because stent spatial geometry is known and unambiguous, reconstruction of endovascular stents should be one of the tests of imaging systems designed to perform three-dimensional reconstruction of ultrasound images. PMID- 8227391 TI - Vascular signals from pleura-based lung lesions studied with pulsed Doppler ultrasonography. AB - Vascular signals arising from pleura-based lung lesions were studied in 50 patients using pulsed Doppler ultrasonography. Twenty-seven had malignant lesions and 23 benign lesions. In 24/27 of the malignant masses, we observed no Doppler signals: in 2 cases, a low-velocity, turbulent, pulsatile flow was demonstrated; in 1 case pulsed Doppler ultrasonography showed only a low-velocity, continuous flow. In 20/23 of the benign lesions, we found 2 types of Doppler signal waveforms: an irregularly pulsatile, venous-like signal, and a regularly pulsatile, arterial-like waveform. The last 8 patients of our series underwent color Doppler ultrasound examination, which demonstrated the presence of arterial and venous vessels in 4 benign lesions and the absence of blood flow in 4 malignant masses. To our knowledge, we report for the first time the ability of obtaining Doppler signals from a variety of lung lesions. The actual clinical relevance of this application requires further studies. PMID- 8227392 TI - Factors affecting image quality and diagnostic efficacy in abdominal sonography: a prospective study of 140 patients. AB - A study was undertaken to evaluate the limitations of abdominal sonography in a group of predominantly elderly patients. In the majority of patients (98%) sonography, using 3.5 MHz and 5 MHz array transducers permitted visualization of the major abdominal organs with image quality sufficient for definitive diagnosis, but improved image quality would be desirable in 78% of this sample. Obesity and immobilization were associated with poor sonographic image quality. In 31 of the 45 (69%) poor image quality studies, no near-field artifacts (less than 3 cm depth) were observed. Obstacles in the acoustic path were noted in 77% of the poor image quality studies. PMID- 8227393 TI - Myeloid metaplasia complicating polycythemia vera: ultrasonographic findings in the stomach. PMID- 8227394 TI - Antenatal ultrasound diagnosis of cerebellar vermian agenesis in a case of rhombencephalosynapsis. PMID- 8227395 TI - Metastatic carcinoid tumor in the liver. PMID- 8227396 TI - Fetal grasping of the umbilical cord causing variable fetal heart rate decelerations. PMID- 8227397 TI - In utero diagnosis of spontaneous fetal urinary bladder rupture. PMID- 8227398 TI - Fetal ultrasonographic detection of hypodontia in the Neu-Laxova syndrome. PMID- 8227399 TI - Ultrasound evaluation of unusual pelvic cystic masses. PMID- 8227400 TI - Update on special techniques in routine cytopathology. PMID- 8227401 TI - Bodies associated with fires. PMID- 8227402 TI - Cost of gentamicin assays carried out by microbiology laboratories. AB - AIMS: To assess the current range of prices charged for gentamicin assays in United Kingdom laboratories; and to examine the laboratories' likely response to increases or decreases in the demand for the service. METHODS: A postal survey of the 420 members of the Association of Medical Microbiologists was used to establish the range of prices charged for aminoglycoside assays. Additionally, eight private institutions were contacted to determine what the private sector was charging for aminoglycoside assays. Reagent costs in the NHS laboratories were calculated by dividing the total cost of all aminoglycoside assay kits by the number of samples analysed. RESULTS: The NHS and the private institutions both showed a wide price variation. Prices charged to an in-hospital requester for a peak and trough assay ranged from 5.00 pounds to 68.20 pounds (n = 44), and to an external private hospital, under a bulk service contract, from 5.00 pounds to 96.00 pounds (n = 47). Prices charged by private laboratories ranged from 49.00 pounds to 84.00 pounds (n = 8). There was a log linear correlation in the NHS laboratories between the reagent costs per assay and the number of assays performed per year, and most laboratories thought that their price per assay would be sensitive to increases or decreases in demand. Laboratories which had purchased their assay machines had lower reagent costs per assay but higher repair and maintenance costs. Overall, number of assays performed and method of payment for assay machinery only accounted for 44.8% of the observed variation in assay kit costs. CONCLUSIONS: The price range for gentamicin assays in the United Kingdom is wide and is only partially explained by the number of assays performed. Most laboratories believe that they would experience a reduction in unit cost as output increases. The currently offered range of prices is, in part, due to variation in the laboratories' approach to costing the service provided and some laboratories charge prices which do not even cover the cost of assay kits. Overall, we believe that prices charged should be as close as possible to the marginal cost of the tests performed. PMID- 8227403 TI - Mandatory use of confirmation stage with Chlamydiazyme during urinary sediment analysis. AB - AIMS: To assess whether false positive results found when the first stage Chlamydiazyme test is performed on urinary sediment could be reduced by using the more specific second stage blocking assay. METHODS: Sediment from 173 urine samples from patients with suspected urinary tract infection caused by Gram negative bacteria and 23 control urine samples were tested using the Chlamydiazyme assay system, which included a blocking assay. RESULTS: A reaction result with the first stage Chlamydiazyme assay test was seen in 102 (58.9%) of the test urine samples. First stage reactivity was not blocked by the Chlamydiazyme confirmatory assay performed on repeat testing. All were correctly identified as true negative (first test false positive) results. CONCLUSIONS: Use of a second (specific) blocking assay for the analysis of urinary sediment using Chlamydiazyme eliminates false positive results in Gram negative urinary tract infections. PMID- 8227404 TI - Electron microscopic changes in Enterocytozoon bieneusi following treatment with albendazole. AB - AIM: To identify and describe electron microscopic changes occurring in Enterocytozoon bieneusi in patients treated with albendazole. METHODS: Eighteen HIV seropositive patients with E bieneusi infection of the small intestine were treated with albendazole 400 mg twice a day for one month. Duodenal biopsy specimens obtained before and at the end of treatment were examined electron microscopically by a pathologist who was unaware of the clinical response. A semiquantitative assessment of the parasite load and description of the parasite morphology was made. RESULTS: A complete resolution of diarrhoea occurred in nine patients and a further three had a greater than 50% reduction in baseline stool frequency or volume. Three of the non-responders were also infected with cryptosporidium. There was a reduction in parasite load in those responding to treatment and an increase in abnormal forms in both responders and non responders. CONCLUSION: The clinical response to albendazole treatment seen in some patients with small intestine microsporidiosis may be due to damage to the developmental stages, causing a partial inhibition of parasite reproduction. PMID- 8227405 TI - Immunological and ultrastructural studies in acute biphenotypic leukaemia. AB - AIMS: To compare the sensitivity of the ultrastructural method to detect myeloperoxidase (MPO) with light microscopy and immunocytochemistry using an anti MPO antibody; to examine the expression of lymphoid antigens in relation to MPO activity in blast cells from cases of biphenotypic leukaemia. METHODS: Blast cells from 14 cases of biphenotypic acute leukaemia were analysed. Immunological markers were performed by single or double immunofluorescence staining on a flow cytometer. The presence of MPO was determined by light microscopy, electron microscopy on fixed and unfixed cells, and by immunoalkaline phosphatase with an anti-MPO antibody. The immunogold method was applied at the ultrastructural level to assess the expression of lymphoid and myeloid antigens at the same time as the MPO activity. RESULTS: Six of the 14 cases were initially classified as acute lymphoblastic leukaemia (ALL) and eight as acute myeloid leukaemia (AML). MPO activity was shown at the ultrastructural level in 4-99% blasts from all cases. Six of the 14 were MPO negative by light microscopy and three of these were negative with the antibody anti-MPO. Coexpression of lymphoid antigens (CD19, CD10, or CD2) and MPO was shown by the immunogold method in four out of 11 cases; in seven cases the blasts coexpressed myeloid antigens (CD13, CD33) and MPO. CONCLUSIONS: Electron microscopy is more sensitive for showing MPO than light microscopy and immunocytochemistry; the immunogold method combined with MPO used at the ultrastructural level can help to define the cell lineage involved in biphenotypic leukaemia by highlighting the myeloid component defined by MPO. PMID- 8227406 TI - Role of beta 2-glycoprotein I and anti-phospholipid antibodies in activation of protein C in vitro. AB - AIMS: To investigate the effect of beta 2-glycoprotein I (beta 2 GPI) on the thrombin/thrombomodulin dependent activation of protein C; and to determine whether beta 2 GPI dependent anticardiolipin antibodies have any effect. METHODS: Protein C was activated by thrombin in the presence of thrombomodulin and phospholipid vesicles in an in vitro system. The effect of adding purified beta 2 GPI to this system was observed. Affinity purified anticardiolipin antibodies and total IgG from patients with anticardiolipin antibodies and the lupus anticoagulant were studied for their effects on protein C activation in the presence and absence of beta 2 GPI. RESULTS: beta 2-Glycoprotein I had no effect on the activity of preformed activated protein C. When the phospholipid vesicles were incubated with beta 2 GPI before the addition of protein C, the activation of protein C was inhibited in a dose dependent manner. With phosphatidylserine:phosphatidylcholine vesicles at a concentration of 1 microM:2 microM, beta 2 GPI began to inhibit the reaction at a concentration of 15 nM, and at 4 microM (the normal plasma concentration) the activation of protein C was reduced to 40%. Anticardiolipin antibodies had no demonstrable effect. CONCLUSIONS: beta 2-Glycoprotein I inhibits protein C activation in an in vitro system. Its physiological role is unknown but it has potential procoagulant as well as anticoagulant properties. An effect of antiphospholipid antibodies on protein C activation, which might explain their association with thrombosis, could not be shown. PMID- 8227407 TI - Evaluation of sensitivity of 10 diagnostic assays for Chlamydia trachomatis by use of a simple laboratory procedure. AB - AIMS: To determine the sensitivity of commercially available diagnostic assays for Chlamydia trachomatis using a simple method. METHODS: Nine commercial assays and an "in-house" polymerase chain reaction (PCR) were evaluated using serial dilutions of a laboratory grown H serovar--four of them using a laboratory grown E serovar. Seven of the assays were further tested using dilutions of several cervical samples known to contain chlamydiae. RESULTS: The most sensitive assays were the MicroTrak direct fluorescent antibody (DFA) test (Syva) and the PCR which detected C trachomatis at a 10(-8) dilution of the H serovar, while the two least sensitive, Clearview (Unipath) and TestPack (Abbott), were positive only at 10(-4) and 10(-3) dilutions, respectively. A range of enzyme immunoassays (EIAs) and a nucleic acid hybridisation test were of intermediate sensitivity. The results with serovar E were consistent with these. When clinical samples were examined, the DFA test detected C trachomatis in dilutions at least 10-fold greater than any other assay. CONCLUSIONS: The range of sensitivity of diagnostic assays determined by the laboratory dilution procedure is very wide. Sensitivity assessed in this way, however, reflects the ability of the assays to detect C trachomatis in large scale clinical trials. The dilution procedure, which is simple to undertake, could therefore be applied by any laboratory before a new diagnostic method is considered for routine use. PMID- 8227408 TI - Haemopoietic regrowth after chemotherapy for acute leukaemia: an immunohistochemical study of bone marrow trephine biopsy specimens. AB - AIM: To analyse haemopoietic regrowth and residual disease in bone marrow trephine biopsy specimens after treatment for acute leukaemia, using immunohistochemical staining. METHODS: Biopsy specimens before and after treatment were studied from patients diagnosed as having acute myeloid or lymphoblastic leukaemia. Specimens after treatment encompassed periods from two to 56 weeks from the start of treatment. Routine haematoxylin and eosin and Giemsa stained sections were evaluated in association with immunostained preparations. A panel of antibodies was used, which reacts with epitopes showing restricted expression dependent on the lineage or maturation stage of cells. Results were evaluated in the light of clinical, peripheral blood, and marrow aspirate findings. RESULTS: The speed and sequence of regrowth of haemopoietic cells were more variable than expected. Immunostaining highlighted features of dysplasia after treatment and in some cases assisted detection of residual or relapsed leukaemia. Peripheral blood and aspirate cell counts reflected accurately the amount of regrowth, but not the dysplasia, seen in biopsy samples. Delayed regrowth was associated with complex individual factors. CONCLUSIONS: Morphological and immunohistochemical study of trephine biopsy specimens from patients treated for acute leukaemia provides information complementary to that obtained from peripheral blood and aspirated marrow. Variation in the timing and sequence of regrowth is highlighted. Immunostaining can aid in the detection of relapse or minimal residual leukaemia. The clinical relevance of dysplastic changes in biopsy specimens after treatment is uncertain, but such changes may persist for long periods. PMID- 8227409 TI - Expression of MUC2 epithelial mucin in breast carcinoma. AB - AIMS: To examine the expression of the MUC2 epithelial mucin in breast carcinoma; to relate this to patient survival. METHODS: Sections from 210 breast carcinomas were stained with the anti-MUC2 core protein monoclonal antibody, 4F1, using an immunoperoxidase technique. The proportion of tumour cells positively stained and the localisation and intensity of any staining were recorded. Expression of MUC2 was compared with histological type and grade, tumour size, presence of nodal metastases, presence of oestrogen receptors, and menopausal status. The prognostic value of MUC2 expression was examined using Kaplan-Meier survival analysis. RESULTS: MUC2 mucin was detected in 19% of cases of invasive carcinoma, in 11% of cases of carcinoma in situ, where present, but very rarely in adjacent normal breast epithelium. Presence of MUC2 was significantly associated with a shorter disease free interval (p < 0.05), although the observed difference in duration of overall survival was not significant. CONCLUSIONS: The MUC2 detected in breast carcinoma may be underglycosylated or staining may represent detection of the protein core before the completion of glycosylation. The virtual absence of 4F1 reactivity in normal breast epithelium suggests that, unlike the MUC1 mucin, the MUC2 mucin is not highly expressed by these cells. The mechanism by which expression of MUC2 affects the biology of breast tumours is unclear, although expression may be a reflection of general derepression of genes during tumour progression. PMID- 8227410 TI - Distribution of gamma delta T-cells in the bronchial tree of smokers and non smokers. AB - AIMS: To assess the distribution of gamma delta T-cells in the human bronchial tree; and to compare quantitatively the differences between gamma delta T-cell numbers in different parts of the airway wall in smokers and non-smokers. METHODS: Full thickness bronchial wall sections were taken from 10 whole lung specimens from both smokers and non-smokers. Serial cryostat sections stained with the monoclonal antibodies CD3 and TCR delta-1 were examined with the aid of interactive image analysis to assess gamma delta T-cell numbers both in absolute terms and as a proportion of total T lymphocyte numbers. RESULTS: In all cases gamma delta T-cells were demonstrable throughout the airway wall. Although in absolute terms they tend to occur predominantly in the bronchial epithelium, this seems to reflect higher numbers of T lymphocytes in the epithelium in general compared with the submucosa. No genuine epitheliotropism is evident. Comparison by smoking status shows a significant increase in gamma delta T-cell numbers in the bronchial glands of smokers compared with non-smokers. CONCLUSIONS: gamma delta T-cells form an integral though variable component of the immunocompetent cell population of the human airway in both smokers and non-smokers. Although epitheliotropism does not exist in the bronchial tree, gamma delta T-cells seem to form a significant part of the bronchial gland inflammation associated with smoking. PMID- 8227411 TI - Epstein-Barr virus in normal, pre-malignant, and malignant lesions of the uterine cervix. AB - AIM--To detect the presence or absence of Epstein-Barr virus (EBV) in cervical lesions ranging from normality to invasive malignancy. METHODS--Eighteen randomly selected cases of invasive squamous cell carcinomas of the uterine cervix were examined as well as 25 cases each of normal cervices and those showing cervical intra-epithelial neoplasia (CIN) I, II, and III. DNA-DNA in situ hybridisation, using a biotinylated probe to the Bam H1 "W" fragment of EBV, was carried out in addition to the polymerase chain reaction using specific primer sequences that flank a 153 base pair segment of the Bam H1 "W" region of the EBV genome and which do not cross-amplify other DNA herpes viruses. Positive control material included paraffin wax embedded P3 HR1 lymphoblastoid cells (containing high copy numbers of EBV) and two nasopharyngeal carcinomas positive for EBV. RESULTS- Neither normal nor CIN I tissue was positive. Eight per cent of CIN II tissue was positive; 8% of CIN III, and 43% of carcinomas were positive for EBV. CONCLUSION- The study shows that the virus is present in some cases of cervical carcinoma and to a lesser degree in some premalignant lesions of the cervix, but the exact association between it and cervical oncogenesis, be it causative or incidental, remains to be determined. PMID- 8227412 TI - Detection of human papillomavirus infection by non-isotopic in situ hybridisation in condylomatous and CIN lesions. AB - AIMS: To study the value of non-isotopic in situ hybridisation (NISH) in detecting human papillomavirus (HPV) infection in female genital lesions positive for the virus by conventional histology but negative by filter DNA hybridisation. METHODS: Forty three cases, which showed the histological hallmarks of the HPV infection but produced negative results in filter dot blot hybridisation tests (Vira Pap and Vira Type kits), were identified in the course of an investigation of 304 vaginal, vulvar, and cervical samples from 267 patients. These cases were studied by NISH for the presence of HPV infection. RESULTS: In 28 (65%) of the cases NISH gave a positive hybridisation signal. In 26 cases (96%) the signal was diffuse, and in two (4%) punctate or diffuse, representing episomal and episomal or integrated HPV DNA, respectively. In most cases only a few HPV positive cells were discernible. CONCLUSION: NISH is a more sensitive technique than dot blot hybridisation, detecting HPV infection in most cases which show histological HPV atypia but which remain negative in filter DNA hybridisation. Thus NISH is useful as an additional technique to verify the presence of HPV in lesions which remain negative in filter hybridisation tests. PMID- 8227413 TI - Detection of immunoglobulin heavy chain gene rearrangements in Hodgkin's disease using PCR. AB - AIM: To detect clonal rearrangements of the immunoglobulin (Ig) heavy chain gene in Hodgkin's disease tissue using the polymerase chain reaction (PCR). METHODS: DNA extracted from 36 samples of Hodgkin's disease was analysed using PCR and primers from conserved sequences in the variable (VH) and joining (JH) regions. RESULTS: Clonal rearrangement was detected only in one case. Evidence of clonal immunoglobulin gene rearrangement had been detected previously in this case using conventional Southern blot analysis. CONCLUSIONS: The sensitivity of the two techniques is equivalent and clonal Ig heavy chain gene rearrangements are rare in Hodgkin's disease. PMID- 8227414 TI - Nucleolar organiser regions and survival in patients with non-Hodgkin's lymphomas classified by the working formulation. AB - AIMS: To correlate the numbers of silver staining nuclear organiser regions (AgNORs) in non-Hodgkin's lymphoma classified by the working formulation with survival, the first complete remission, and the length of remission. METHODS: Sixty one patients were included in the study. Paraffin wax sections were stained using silver solution to visualise nucleolar organiser regions. The AgNORs were counted in 150 nuclei of each specimen. Data were examined using the Kruskal Wallis test, multivariate discriminant analysis, and Cox's regression test. Curves were calculated by the method of Kaplan and Meier. RESULTS: Most patients who were alive had low grade lymphoma (p < 0.01). The first complete remission was obtained more frequently in the low and intermediate grade groups (p < 0.05). The longest survival was found in the low grade group (p < 0.001). The mean number of AgNORs differed significantly in all three groups (p < 0.001). This was also true for area of nuclei (p < 0.001) and length of remission (p < 0.05). In a multivariate analysis the numbers of AgNORs were highly predictive for survival, remission, and the length of remission. CONCLUSIONS: The numbers of AgNORs correlated with survival, remission, and the length of remission in patients with non-Hodgkin's lymphoma. PMID- 8227415 TI - Prognostic importance of DNA flow cytometric variables in rhabdomyosarcomas. AB - AIM: To determine whether DNA ploidy patterns and S phase fraction offer prognostic information in patients with rhabdomyosarcoma (RMS). METHODS: DNA flow cytometry was performed on formalin fixed, paraffin wax embedded samples from primary tumours, and metastatic deposits or recurrences in 70 patients. DNA histogram analysis was done using a semi-automated cell cycle analysis program. RESULTS: Of the 70 primary tumours, 23 were DNA diploid, 32 DNA aneuploid, eight DNA multiploid, and seven DNA tetraploid. The prognosis for DNA aneuploid patterns was favourable, intermediate within the group of DNA tetraploid tumours and poor among patients with DNA diploid and DNA multiploid tumours (p = 0.009). In multivariate analysis (Cox regression model) DNA ploidy was an important independent prognostic factor, along with TNM stage, localisation, and histopathological classification. Ten out of 32 patients with a high S phase fraction (> 15%) with primary RMS achieved long term survival in contrast to 20 out of 29 patients with a low S phase fraction (< or = 15%) (p = 0.008). In 24 cases the DNA ploidy of cases of relapse was analysed. Of the 15 cases, in which stem line changes had occurred, 13 died of disease. No stem line changes were noted in nine cases and in this group four patients died of disease (p = 0.02). CONCLUSIONS: Assessment of DNA ploidy and S phase fraction in primary RMS and evaluation of stem line changes in cases of relapse are important variables in predicting prognosis. PMID- 8227416 TI - Necrotising granulomas of the uterine corpus. AB - Necrotising granulomatous inflammation of the uterine corpus associated with transcervical laser ablation of the endometrium occurred in four patients. The abnormalities seen, including extensive necrosis and hyalinisation with foreign body giant cells containing black foreign material, and eosinophilic homogenisation around blood vessels, were due to the effects of tissue fulguration. The presence of black foreign material in loosely organised histiocytes should alert the pathologist even in the absence of clinical information; stains for organisms are negative. PMID- 8227417 TI - Malakoplakia and adenocarcinoma of the caecum: a rare association. AB - Colonic malakoplakia can occur either as a part of systemic disease with multiple colonic and extracolonic deposits or as an incidental finding associated with a rectal or sigmoidal adenocarcinoma. A case is reported which shows an association between malakoplakia and adenocarcinoma of the caecum. PMID- 8227418 TI - Mesenchymal chondrosarcoma arising in fibrous dysplasia of the femur. AB - The occurrence of mesenchymal chondrosarcoma in an area of fibrous dysplasia in the upper femur of a 28 year old man is reported. It is believed that this is the first documented example of such an association. A further unusual feature is the presence of benign giant cells in the mesenchymal chondrosarcomatous component. PMID- 8227419 TI - Hypochromic macrocytes: are they reticulocytes? AB - Automated full blood counters measuring the size and haemoglobin concentration of individual red cells by high and low angle light scatter can provide estimates of the percentage of cells which are abnormal in either or both of these variables. The hypothesis that an increase in the number of large cells with a reduced haemoglobin content ("hypochromic macrocytes") was indicative of a reticulocytosis was investigated. A correlation was shown between the percentage of hypochromic macrocytes and the reticulocyte count. This correlation was too weak to allow the actual reticulocyte percentage to be predicted from the percentage of hypochromic macrocytes. An increased percentage of hypochromic macrocytes, however, often indicates an increased reticulocyte count and thus serves to chart the haematologist to this possibility. PMID- 8227420 TI - Fatal infection associated with group C streptococci. AB - Serious infection caused by Lancefield group C streptococci is unusual in man. Two unrelated deaths associated with these organisms in a 55 year old woman who died after three days of diarrhoea and vomiting, and in a 65 old man who died after a week of non-specific symptoms, are presented. Post mortem examination showed septicaemia in the former and severe aortic stenosis with widespread septic emboli and probable meningitis in the latter. Lancefield group C streptococci were isolated from both cases. These organisms may be carried asymptomatically and usually cause disease in animals but cases of serious human infection have recently been described, mainly in elderly patients or those with other predisposing factors. PMID- 8227421 TI - Bone marrow biopsy specimens in assessment of remission in acute leukaemias. PMID- 8227422 TI - Histopathology EQAs: the definition and identification of poor performance. PMID- 8227423 TI - Ethics and necropsies. PMID- 8227424 TI - Value of face masks at post mortem examination. PMID- 8227425 TI - Current thoughts on the pathogenesis of graft versus host disease. PMID- 8227426 TI - ACP Broadsheet No 141: September 1993. Role of endocrine biochemistry laboratories in the investigation of infertility. AB - The staff and services of the endocrine biochemistry laboratory are essential to the efficient investigation of infertility. Each centre should adopt a detailed strategy for the investigation of the infertile couple which specifies the hormone analyses required at each stage. Appropriate first-line hormone tests should be selected after a thorough clinical history and physical examination of both partners. Second-line hormone testing should be determined from the results of the initial investigation and should be restricted to requests that either confirm or clarify an endocrine basis to infertility or monitor the response to treatment. The clinical biochemist should advise on specimen timing and collection, have responsibility for guaranteeing time and valid hormone results, and be part of the team that audits the overall strategy and the outcome for individual patients. PMID- 8227427 TI - Computed cell cycle and DNA histogram analyses in image cytometry in breast cancer. AB - AIMS: To analyse the cell cycle and DNA histogram components in data from DNA static cytometry and, in particular, to investigate the influence of the length of time the slides are exposed to the light of the cytophotometer in evaluating the G0/G1 peak. METHODS: DNA static cytometry was performed on 18 Feulgen stained imprints and six histological sections taken from six breast carcinomas. The total optical density values obtained were analysed using software commercially available as Multicycle. DNA flow cytometry was performed on the same cases. RESULTS: The proportions of nuclei related to the cell cycle components from DNA static cytometric data, obtained from Feulgen stained cytological smears, were almost identical with those obtained from DNA flow cytometric data. Moreover, additional information was obtained from the DNA static cytometry frequency histogram and the proportions of nuclei below the diploid G0/G1 peak and above the G2 phase. Discrepancies between DNA static cytometry and DNA flow cytometry were seen in the large coefficients of variation of the G0/G1 peaks obtained with the former method of analysis, even though a better correspondence was found when the exposure time of the slides to the light of the cytophometer was conspicuously shortened. The information obtained from histological sections seemed to be similar to that obtained from DNA flow cytometry when a single cell population was present; a single cell population was detected in two out of the three cases in which two distinct populations had been present in DNA flow cytometry. CONCLUSIONS: The computer analysis of DNA static cytometric data obtained from Feulgen stained cytological specimens provides the type of information on the cell cycle which is usually obtainable only from DNA flow cytometry. Correspondence with the DNA data from histological sections, however, was poor. PMID- 8227428 TI - Wide needle aspiration cytology in the diagnosis of lymphadenopathy in Zambia. AB - AIMS: To study the value of wide needle (19 gauge) aspiration cytology in the diagnosis of lymph node disease in Zambia in the absence of a trained cytologist. METHODS: Patients (n = 304) referred for surgical biopsy of an enlarged peripheral lymph node were studied prospectively. Surgical biopsy was routinely preceded by 19 gauge needle aspiration of the same node; aspirates were stained by haematoxylin and eosin and Ziehl Neelsen stains. RESULTS: Of 232 aspirates, 182 contained sufficient material for cytological characterisation. Tuberculosis was diagnosed or suspected in 122 of 126 aspirates with histologically confirmed tuberculous lymphadenitis; reactive follicular hyperplasia in 31 of 38 patients with primary HIV lymphadenopathy; malignancy in all five patients with malignant nodes; and Kaposi's disease in four of nine patients with this. Tuberculous lymphadenitis was falsely suspected in four patients, as was reactive follicular hyperplasia in four, and Kaposi's disease in four. CONCLUSIONS: Wide needle aspiration cytology is useful in the diagnosis of lymphadenopathy in Central Africa, with the exception of lymphadenopathic Kaposi's disease. PMID- 8227429 TI - Proliferative activity of normal and neoplastic urothelium and its relation to epidermal growth factor and transferrin receptors. AB - AIMS: To investigate the proliferative activity (given by the Ki67 index) of the normal, atypical, and neoplastic urothelium and its relation to the cellular reactivity for the epidermal growth factor (EGFr) and transferrin (Tfr) receptors. METHODS: The Ki67 index and the level of EGFr and Tfr reactivity were determined on frozen sections from 82 patients with urothelial cancer. Relevant clinical material was reviewed to establish correlations with the degree of atypia and invasion. RESULTS: Morphologically normal urothelium, whether derived from controls or patients with cancer, exhibited a low Ki67 index (less than 0.1%) and weak receptor reactivity. In transitional cell carcinomas (TCCs) the Ki67 index was increased: it ranged between 0.7% and 10% in non-invasive and exceeded 10% in 88% of the invasive TCCs. Strong positive reactions for EGFr were seen only in invasive TCCs, but in 47% of invasive TCCs the EGFr was not "overexpressed" and did not match the Ki67 index. A better correlation was found between the Ki67 index and the Tfr which was positive in 26% of the non-invasive and in 71% of the invasive tumours. All three variables were increased in severe atypia but varied considerably in lesser degrees of atypia. CONCLUSIONS: Despite the absence of a close correlation, accelerated growth and enhanced receptor expression were characteristic of invasive cancers. These results suggest that the growth rate in TCCs is not causally related to overexpression of growth factor receptors but that the latter is an abnormality which may accompany the malignant phenotype. PMID- 8227430 TI - Evaluation of Questor urine screening system for bacteriuria and pyuria. AB - AIMS: To evaluate the Questor automated bacteriuria and pyuria screening system; to compare its performance with that of a reference method; and to assess its usefulness in a routine clinical laboratory. METHODS: The Questor urine screening system was compared with a comprehensive regimen to detect urinary tract infection, using pour-plate viable counts to determine the numbers of bacteria present in urine samples, a wide range of other cultural methods, microscopic findings and clinical information. RESULTS: The optimal performance in detecting significant growths was a sensitivity of 93%, a specificity of 74%, a positive predictive value of 43% and a negative predictive value of 98%. The list price per test is 0.17 pounds and the capital cost of the system is 39,950 pounds. Questor can test 50 samples an hour and can be operated by one member of the laboratory staff, who is not required to make interpretative judgments--for example, a medical laboratory assistant. CONCLUSIONS: The sensitivity and specificity of the Questor was better than that obtained from other screening systems using the same protocol. The system was easy to use and is a useful addition to the methods available for screening for bacteriuria. PMID- 8227431 TI - Use of a commercial PCR kit for detecting Chlamydia trachomatis. AB - AIMS: To evaluate a commercial polymerase chain reaction (PCR) kit for the detection of Chlamydia trachomatis. METHODS: Two hundred and fifty seven genital specimens, which had been submitted in 2SP medium for chlamydial isolation and subsequently stored at -70 degrees C, were retrospectively examined by a commercial PCR kit which detects chlamydial plasmid DNA. Culture negative, PCR positive specimens were examined by immunofluorescence and an in-house major outer membrane protein (MOMP)-PCR. RESULTS: All 49 specimens which were culture positive were also PCR positive. Another 14 specimens were also PCR positive. After resolution of these results by immunofluorescence and a PCR assay for MOMP the sensitivity for PCR was 98.4% and that of culture 79%. The specificities were 99.5% and 100%, respectively. CONCLUSIONS: This kit, which is highly sensitive and specific, is straightforward to use and has a built-in safeguard against cross contamination. The role of this test in the examination of routine genital specimens from patients with uncomplicated chlamydial infection is questionable due to its expense. It may have a place in the investigation of trachoma or infertility, however, where it has been shown that DNA can be detected when culture is unsuccessful. PMID- 8227432 TI - Serum pepsinogen I and II concentrations and IgG antibody to Helicobacter pylori in dyspeptic patients. AB - AIMS: To investigate the association between histologically confirmed gastritis, carriage of Helicobacter pylori and pepsinogen (PG) I and PG II concentrations. METHODS: Prospective study of 81 dyspeptic patients undergoing upper gastrointestinal endoscopy was made. The extent of gastric mucosal inflammation and the presence of H pylori was determined, and serology to evaluate PG I and II concentrations and IgG titres to H pylori was carried out. RESULTS: The presence of H pylori was strongly correlated with high IgG antibody titres to H pylori and gastritis. Patients who were H pylori positive had significantly higher PG I and PG II concentrations and a significantly lower PG I:PG II ratio than patients who were negative for H pylori. In 13 patients with duodenal ulcer and H pylori positive gastritis serum PG I concentrations were significantly higher than in H pylori positive patients without duodenal ulcer. Significant correlations were found between the age of patients and serum PG II, the PG I:PG II ratio, IgG antibodies to H pylori, the severity of body gastritis and H pylori infection, and between the degree of gastritis in the body of the stomach and the PG II concentration. CONCLUSIONS: Serum PG I and II concentrations, together with a fall in the PG I:PG II ratio, could be used as predictors of H pylori infection as well as serum IgG antibody response to H pylori. PMID- 8227433 TI - Selective medium for thermophilic campylobacters including Campylobacter upsaliensis. AB - AIMS: To develop a selective supplement for the recovery of thermophilic Campylobacter spp, including Campylobacter upsaliensis from faeces, using campylobacter blood free selective agar base as the growth medium. METHODS: Minimum inhibitory concentrations (MICs) of cefoperazone and of teicoplanin were determined for 51 strains of C upsaliensis, 159 strains of other thermophilic Campylobacter spp, and for 66 Enterobacteriaceae (cefoperazone only). From these results a medium using campylobacter blood free selective agar base incorporating cefoperazone (8 mg/l), amphotericin (10 mg/l), and teicoplanin (4 mg/l)--CAT medium--was formulated and compared with the commercially available campylobacter blood free selective medium (modified CCDA) for isolation of Campylobacter spp from 7000 human faecal specimens. The two media were also compared for the recovery of C upsaliensis from 45 spiked human faeces. RESULTS: Isolation rates of Campylobacter spp other than C upsaliensis were similar for both media, but the CAT medium alone recovered four of the five strains of C upsaliensis from the faecal samples examined. From the spiked faeces specimens, recovery of C upsaliensis was increased by between 35% and over 200-fold on the CAT medium compared with the modified CCDA. CONCLUSIONS: CAT selective agar was a suitable alternative medium to modified CCDA for the growth of thermophilic Campylobacter spp, including C upsaliensis from faeces. PMID- 8227434 TI - Prevalence of lymphoid follicles and aggregates in Helicobacter pylori gastritis in antral and body mucosa. AB - AIMS: To evaluate the prevalence of lymphoid follicles and aggregates in the antral and body mucosa in Helicobacter pylori gastritis and to assess if there were correlations with ulcers in the duodenum, pylorus, or stomach, and with chronic antral erosions. METHODS: Patients (n = 2692) with histologically confirmed H pylori antral gastritis were investigated. These comprised five groups: those with duodenal ulcers; those with pyloric ulcers; those with gastric ulcers; those with chronic erosions; and those with no associated lesions. In 1446 cases at least two additional biopsy specimens from the oxyntic mucosa were available. RESULTS: Lymphoid follicles and aggregates were found in 53.8% of cases in the antral mucosa compared with 14.8% in the oxyntic mucosa (p < 0.001). The various diseases showed significant differences in terms of the prevalence of follicles and aggregates: The highest numbers in the antral mucosa as well as the lowest in the oxyntic mucosa were found in patients with duodenal ulcers (60.5% and 9.2%, respectively). The highest numbers of follicles and aggregates in the oxyntic mucosa occurred in patients with gastric ulcers. CONCLUSIONS: The detection of lymphoid follicles and aggregates in oxyntic mucosa and the higher prevalence in antral mucosa fits well with the distribution of primary gastric lymphomas. This adds further weight to the notion that the development of follicles and aggregates, triggered by H pylori, might be an early precursor to gastric lymphoma. The differences between the groups investigated might be due to different strains of H pylori or differences in the respective sizes of antral and oxyntic mucosa. PMID- 8227435 TI - Prostaglandin E2 in gastric mucosa of children with Helicobacter pylori gastritis: relation to thickness of mucus gel layer. AB - AIMS: To evaluate the changes in mucus gel layer thickness and prostaglandin E2 (PGE2) content caused by Helicobacter pylori infection in the antral mucosa of children: to assess whether decreased mucus gel thickness is related to PGE2 production. METHODS: Antral biopsy specimens were taken at endoscopy from 153 children. H pylori gastritis was evident in 45 and normal mucosa in 59. The other 49 children were studied one month after antibiotic treatment that eradicated the infection in 37 of them had been stopped. One antral specimen was immersed in ice cold saline, put under an inverse microscope with an eyepiece graticule. Mucus gel thickness was measured and then the processed for histological examination; another specimen was weighed and processed for in vitro prostanoid generation. RESULTS: Mucus gel layer thickness was significantly decreased in children with H pylori gastritis (90 (SD) 29) microns v 120 (58) microns in controls, p < 0.01) but returned to control values after H pylori had been eradicated. PGE2 generation was significantly increased in children with H pylori gastritis (1022 (811) ng/g v 641 (473) ng/g in controls, p < 0.01). One month after treatment PGE2 generation significantly decreased in children without infection (880 (534), p < 0.01), but was still high where infection persisted. A significant inverse correlation was found between PGE2 generation and mucus gel layer thickness (p < 0.05). CONCLUSIONS: These data suggest that H pylori damages the mucus gel layer, and that the gastric mucosa increases generation of PGE2 in response to back diffusion of acid and pepsin. PMID- 8227436 TI - Use of recombinant human parvovirus B19 antigens in serological assays. AB - AIMS--To compare the sensitivity, specificity, and practicality of recombinant proteins in serological tests for the detection of human parvovirus B19 IgG and IgM. METHODS--Indirect enzyme linked immunosorbent assays using B19 structural proteins expressed in Escherichia coli were developed for the detection of B19 specific IgG and IgM (rELISA-G and rELISA-M). Cells infected with baculovirus expressing B19 structural proteins were also used in an indirect immunofluorescence assay for IgG and IgM antibodies (IFA-G and IFA-M). Antibody capture radioimmunoassays for IgG and IgM (GACRIA and MACRIA) were used as comparative assays. RESULTS--Twenty nine pools of intravenous immunoglobulin were clearly positive for B19 IgG by rELISA-G and contained low IgG titres by GACRIA. From 113 samples tested by all methods, sensitivities of 92% (77/84) and 97% (68/70) were obtained for ELISA and immunofluorescence, respectively, when compared with GACRIA. One hundred and sixteen samples from patients presenting with rash or arthralgia were compared by MACRIA, rELISA-M, and IFA-M. Sensitivities of both recombinant tests were more than 95%. Despite pretreatment to remove IgG or rheumatoid factor, false positive results were a problem in the rELISA-M but were not seen with the IFA-M. CONCLUSIONS--The limited supply of native antigen has severely restricted the wide application of serology for parvovirus B19. The use of recombinant antigens permitted the introduction of local screening tests which had many advantages, including quicker results and relief of the burden on the Reference Laboratory. The use of rELISA-M for sensitivity and IFA-M for specificity and confirmation proved a useful and practical combination for diagnosis of recent infection with B19, and rELISA-G allowed the immune response to be determined in selected populations. PMID- 8227437 TI - Epstein-Barr virus in carcinoma of the vulva. AB - AIMS: To detect the presence of Epstein-Barr virus (EBV) in cases of vulval carcinoma in Chinese patients living in Hong Kong. METHODS: Formalin fixed, paraffin wax embedded blocks from eight cases of vulval carcinoma and six age matched controls of non-neoplastic vulval tissue were analysed for the presence of EBV DNA using the polymerase chain reaction (PCR). RESULTS: EBV DNA was detected in only one of the eight cases of vulval carcinoma cases while it was detected in four out of the six control cases. CONCLUSIONS: There is no demonstrable association between EBV and vulval carcinoma. Detection of EBV in non-neoplastic vulval epithelium highlights its ubiquitous presence in the lower female genital tract. PMID- 8227438 TI - Use of plasma ferritin concentration to diagnose iron deficiency in elderly patients. AB - AIMS: To determine a concentration of ferritin below which the possibility of iron deficiency should be considered in elderly patients. METHODS: Consecutive new referrals to a geriatric unit (n = 472) were studied prospectively. Full blood count, ferritin, serum vitamin B12 and red cell folate were measured for all patients. A blood film was assessed independently by three haematologists for features of iron deficiency. For those with ferritin of 12-45 ng/ml, bone marrow aspirates were performed and examined for the presence of stainable iron. When possible, a trial of oral iron was given to those with ferritin of < or = 45 ng/ml and response was determined by re-measurement of full blood count and ferritin after a minimum of three weeks of treatment. RESULTS: Bone marrow examination was performed in 32 patients with ferritin of 12-45 ng/ml, of whom 27 (84%) had absent stainable iron, suggesting that most elderly patients with ferritin in this range have iron deficiency. Compared with those with ferritin of 100-299 ng/ml, in whom iron stores were presumed to be normal, patients with ferritin of 12-45 ng/ml had a significantly lower mean haemoglobin and mean red blood cell volume. Furthermore, patients with ferritin up to 75 ng/ml had a significantly higher mean red cell distribution width, and were more likely to have an iron deficient blood film. CONCLUSION: Iron deficient erythropoiesis can occur in elderly patients with ferritin up to 75 ng/ml. This is much higher than the lower limit of the "normal" range usually quoted for younger subjects; this difference should be taken into account when ferritin concentrations are interpreted in elderly patients. PMID- 8227439 TI - Increased pentane and carbon disulfide in the breath of patients with schizophrenia. AB - AIMS: To determine the concentrations of pentane (a marker of lipid peroxidation) and other volatile organic compounds in the breath of patients with schizophrenia. METHODS: Volatile organic compounds were assayed by gas chromatography/mass spectroscopy (GC/MS) in 88 subjects--25 with acute schizophrenic psychosis, 26 with psychiatric disorders other than schizophrenia, and 37 normal volunteers. RESULTS: The mean alveolar gradients of pentane and carbon disulfide (CS2) were significantly higher in the patients with schizophrenia than in the control groups. CONCLUSIONS: Schizophrenia may be accompanied by accelerated lipid peroxidation in cell membranes, as well as increased manufacture of CS2, a known neurotoxin. PMID- 8227440 TI - Fatal case of echovirus type 9 encephalitis. AB - Enteroviruses are rare causes of acute focal encephalitis. A fatal case of echovirus type 9 infection is reported in a 9 month old boy who presented with a fever and a macular rash followed by two focal seizures. Echovirus type 9 was isolated from lung tissue after seven days. PMID- 8227441 TI - Benign cystic peritoneal mesothelioma. AB - The well defined but rare entity of benign cystic mesothelioma is reported. The aetiology of this neoplasm remains obscure. The presenting features make a precise preoperative diagnosis difficult; information provided by computed tomography and cytology may help. A firm diagnosis can only come from electron microscopic or immunohistochemical examination of the tumour. Diagnostic accuracy and diligent follow up are essential because, although the tumour is considered benign, it does tend towards local recurrence. PMID- 8227442 TI - Multiple benign stromal cell tumours of the small bowel. AB - A rare case of multiple small intestinal stromal cell tumours is described in a 79 year old woman who presented with melaena and anaemia. At surgery, about 40 lesions were noted on the serosal surface of the small intestine, the largest of these tumours being located in the mid-jejunum. This lesion showed central necrosis and haemorrhage with a sinus opening into the jejunal lumen. Histological examination of this jejunal tumour showed epithelioid and spindle shaped cells. A smaller biopsied tumour was a pure spindle cell lesion. Both lesions fitted the criteria for benign stromal cell tumours. Although skenoid fibres were identified, the immunophenotype was characteristically heterogeneous. Multiple benign small intestinal tumours can raise the spectre of metastases when seen at surgery, although their presence in this case does not seem to indicate this. PMID- 8227443 TI - Mesenteric lipodystrophy. AB - Three cases of mesenteric lipodystrophy with a wide range of clinicopathological features are reported. Mesenteric lipodystrophy may present as an acute abdomen or with non-specific upper abdominal symptoms. Routine biochemical and haematological investigations are within normal limits. Histological examination shows lipid-filled macrophages in sheets and bands with focal cyst formation. Mesenteric lipodystrophy is a rare condition. A firm diagnosis can be reached only by histological examination and a number of conditions need to be considered in the differential diagnosis. PMID- 8227444 TI - Multinucleated stromal giant cells of the colonic lamina propria in ulcerative colitis. AB - Multinucleated stromal giant cells were seen in the colonic mucosa in biopsy specimens from two patients with long-standing quiescent ulcerative colitis. Similar cells have been described at other sites associated with chronic inflammation, including the lower female genital tract, bladder and anus. The immunophenotype of the cells in the colonic mucosa suggested that they had originated from fibroblasts rather than histiocytes, in common with cells seen at other sites of inflammation. These examples lend support to the concept of there being a reactive morphological change possibly related to interaction with mast cells. These multinucleated giant cells are distinct from histiocytic giant cells and should not be confused with them. PMID- 8227445 TI - Effect of embolisation of meningioma on Ki-67 proliferation index. AB - The proliferation indices (PIs) were determined in 33 meningiomas using two monoclonal antibodies to the Ki-67 antigen (Ki-67 and MIB1). PIs obtained using Ki-67 on frozen material were intermediate between the lowest and highest MIB1 PIs determined in paraffin wax sections. Preoperative embolisation did not influence Ki67 PI, but four of the 15 embolised tumours showed substantially increased MIB1 PIs around embolisation necroses. Proliferation can be reliably assessed in routinely processed meningioma tissues using MIB1; and increased perinecrotic PIs may occur in embolised meningiomas without being an indication of malignancy. PMID- 8227446 TI - Errors in automated reticulocyte counts due to Heinz bodies. AB - Serial reticulocyte counts on a patient with an unstable haemoglobin haemolytic anaemia whose red cells contained Heinz bodies were obtained by two automated methods. One false low and several false high counts were obtained with standard threshold settings; each was accompanied by activation of the appropriate alarm. With the FACScan a bimodal red cell distribution was found, corresponding to the proportions of Heinz body positive and negative cells. This case illustrates the fact that Heinz bodies can interfere with automated reticulocyte counting methods, and to a degree that could be clinically important. PMID- 8227447 TI - Hypophosphatasia: dental aspects and mode of inheritance. AB - Hypophosphatasia is rare enzymopathy that normally presents within the first few years of life and often has profound effects upon the periodontium. It is a heritable disorder characterised by defective mineralisation of the skeletal and dental structures of the body and a deficiency in the liver/bone/kidney (L/B/K) isoenzyme of alkaline phosphatase (ALP). There has been a tremendous advance in our knowledge of this condition over the last decade due to the advent of highly specific DNA probes and novel microanalytical techniques. This paper aims to review current literature about hypophosphatasia with special reference to the dental aspects of the condition and to shed light upon the controversial area of its mode of genetic inheritance. It is concluded that hypophosphatasia may result from the existence of 2 defective alleles, which alone or in combination may cause the condition. One allele is expressed in an autosomal dominant manner producing milder phenotypic characteristics, whilst the other is expressed in an autosomal recessive manner producing the more severe clinical form that often results in neonatal death. The milder phenotypes may go undiagnosed and the consequence of this in genetic counselling terms is extremely important. PMID- 8227448 TI - Probing depth at implants and teeth. An experimental study in the dog. AB - The aim of the present investigation was to assess the resistance offered by the gingiva at teeth and the peri-implant mucosa at osseointegrated titanium implants to mechanical probing. 5 beagle dogs were used in the experiment. The 2nd and 3rd premolars and the 1st molars of the right and left mandibular dentition were extracted. 2 titanium implants were installed in the edentulous right and left premolar- and molar-regions. Abutment connection was performed 3 months later. The remaining premolar in the left jaw (P4) was exposed to "experimental periodontitis" during a 4-month period and, thus, the 4th premolar in the right jaw (4P) was representing healthy periodontium. The inflamed soft tissues at P4 were treated using a flap procedure. Following 2 weeks of healing, the main experiment was started, i.e., day 0. During the subsequent 360 days, the teeth and abutment parts of the implants were regularly exposed to plaque control (3 x/week). Radiographs of 4P-, P4- and the implant-regions were obtained on days 0 and 360. Mobility measurements were performed on days 0 and 360. Biopsies of the experimental teeth and the implants were sampled at the end of the study. The results of the present experiment demonstrated that differences in terms of tissue composition, organization and attachment between the gingiva and the root surface on one hand and between the peri-implant mucosa and the implant surface on the other, make the conditions for probing depth measurements at teeth and implants different.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227449 TI - One-year trial of the effect of rinsing with an amine fluoride-stannous-fluoride containing mouthwash on gingival index scores and salivary microbial counts in lymphoma patients receiving cytostatic drugs. AB - This double-blind study was undertaken to evaluate the effect of twice-daily use of a mouthwash containing 0.025% of fluoride as amine fluoride-stannous fluoride (AmF + SnF) or 0.05% of fluoride as sodium fluoride (NaF) on visible plaque index (VPI) and gingival bleeding index (GBI) and on some salivary micro-organisms in patients suffering from non-Hodgkin or Hodgkin lymphomas. 79 patients were allocated at random to the two mouthwash groups. Mouthwashing began at the start of cancer chemotherapy. Results relating to 45 patients who completed a 1-year rinsing protocol showed a significant decrease in VPI and GBI in the AmF + SnF group. An increase was found in the NaF group. Mean values for stimulated salivary secretion rates and buffering capacities mostly did not differ significantly from baseline values during the study. In both groups, mutans streptococci counts decreased significantly after the study began and remained low in the AmF + SnF group. No corresponding effect was seen in relation to lactobacilli and yeast counts. In the NaF group, lactobacilli counts increased significantly over a year. Significantly more patients reported adverse or unpleasant effects in the AmF + SnF group (52%) than in the NaF group (6%), although both solutions had the same colour and taste. However, all patients continued with rinsing. PMID- 8227450 TI - Efficacy of meclofenamate sodium (Meclomen) in the treatment of rapidly progressive periodontitis. AB - This 6-month, double-blind, controlled clinical trial determined the efficacy of the non-steroidal anti-inflammatory drug, meclofenamate sodium (Meclomen), as an adjunct to scaling and root planing in the treatment of rapidly progressive periodontitis (RPP). 22 subjects (7 male, 15 female) aged 36.5 +/- 7.88 years with RPP and disease-active sites as determined by pretreatment bone scan had standardized radiographs at baseline and 6 months, and clinical measurements at baseline, 3 and 6 months. Following full-mouth scaling and root planing, subjects were randomly assigned to either a placebo, 50 or 100 mg meclofenamate sodium bid group. Bone change over the 6-month period as assessed by subtraction radiography was the primary efficacy determinant. Specialized software was used to isolate the lesion from the subtraction image and to measure bone change along the root surface. ANOVA using the subject as the unit of analysis revealed a significant dose response (p < 0.001) with the placebo group having a mean bone loss of 0.42 +/- 0.06 mm and the low and high dose groups having mean bone gains of 0.07 +/- 0.05 and 0.20 +/- 0.07 mm, respectively. These findings indicate that meclofenamate sodium may be a useful adjunct in the treatment of rapidly progressive periodontitis. PMID- 8227451 TI - Epidemiology and transmission of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans among children and their family members. A report of 4 surveys. AB - The distribution and transmission of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in 4 families were studied. The families were included, based on the isolation of P. gingivalis from a young child or adolescent. The probands of these 4 families were: a 5-year old periodontally healthy boy; a 17 year old girl with severe generalized juvenile periodontitis; an 11-year old girl with prepubertal periodontitis; 2 sisters, 5 and 17-years old, with untreated severe periodontitis as a component of the Papillon-Lefevre syndrome. All members of the 4 families were examined clinically and microbiologically for the presence of P. gingivalis and A. actinomycetemcomitans. Most of the parents appeared to be adult periodontitis patients; the parents of one proband were edentulous. Results showed that in all cases at least one of the parents was positive for P. gingivalis. On the basis of indistinguishable restriction endonuclease patterns (REPs) of P. gingivalis and A. actinomycetemcomitans isolates from parents and their children, and distinct REPs from unrelated individuals, the present study indicates that P. gingivalis and A. actinomycetemcomitans were transmitted between parents and their children. PMID- 8227452 TI - A rapid and simple method for counting crevicular polymorphonuclear leucocytes. AB - The techniques presently available for counting crevicular polymorphonuclear leucocytes (CPMNs) require special equipment and are only semi-quantitative. The present new and simple method is based on the property that Durapore strips have to harvest a maximum number of crevicular cells and to release them when shaken in a suitable buffer. Durapore strips were inserted into periodontal crevices or pockets, shaken in PBS and the liberated PMNs counted in a Neubauer chamber. The validity of the method was ascertained in 5 experiments. First, it was shown that 90% of the harvested cells were released from Durapore strips after being shaken in buffer. Second, decreasing numbers of CPMNs were found when performing sequential samplings. Third, when counting CPMNs before and after therapy, a significant decrease in the number of cells was found. Fourth, we could confirm that sites with bleeding on probing did contain a much higher number of CPMNs as compared to non-bleeding sites. Finally, our counting technique confirmed that deeper pockets contain a significantly higher number of CPMNs. PMID- 8227453 TI - Comparative estimation of periodontal conditions by means of different index systems. AB - The aim of the present study was to compare epidemiological data of periodontal disease obtained from a sample of adults by means of different, commonly employed, partial and full-mouth index systems, in order to explore the amount of discrepancy attributed to the methodology per se. 169 dentate subjects, aged 25 64 years, were subjected to a clinical examination, including circumferential probing assessments of pocket depth (PPD) and attachment level (PAL) at all teeth present. The individual mean % of tooth sites with PPD of > or = 6 mm and the % of subjects exhibiting at least one such deep pocket were calculated based on (i) full-mouth data, (ii) data derived from the buccal and mesial surfaces from 1 randomly selected upper and 1 lower quadrant, (iii) probing assessments at the 6 "Ramfjord teeth", (iv) the full-mouth community periodontal index for treatment needs (CPITN), and (v) the partial CPITN based on 10 index teeth. The PAL data were analyzed by means of 3 versions of the extent and severity index, 1 generated by full-mouth assessments and 2 by partial assessments based on 28 and 10 tooth sites, respectively. In the entire sample, the individual mean % of sites with PPD of > or = 6 mm generated by the different systems ranged between 5.0 and 4.2 sites/subject. By full-mouth CPITN scorings, an average of 1.0 score 4 sextants/subject was recorded, while the partial CPITN generated a corresponding value of 0.8 score-4 sextants/subject.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227454 TI - Late onset Papillon-Lefevre syndrome? A chromosomic, neutrophil function and microbiological study. AB - We had the opportunity to study a family with one of the most destructive forms of periodontal disease known, the Papillon-Lefevre syndrome. The parents had no consanguinity and were not affected, and were therefore to be considered carriers of the disease. 2 sisters, the eldest and youngest, showed periodontal breakdown and hyperkeratotic skin lesions, but their deciduous dentition was not affected. 2 brothers had skin lesions only and another brother and sister were healthy. Furthermore, 2 babies died at birth one after a 9-month pregnancy and the other after a 6-month pregnancy, and the mother also suffered 3 miscarriages. For 4 years, we studied the family: in the case of both sisters, mechanical periodontal treatment and antibiotics were unable to control the disease. In the chromosomic study of the 2 sisters affected, the GTG banding technique found no trace of anomalies in the cells analyzed, whose chromosomic formation was 46,XX. Before treatment, the chemotaxis of the PMN, the phagocytosis of opsonized Staphylococcus aureus, and production of superoxide radicals by PMN was significantly impaired in both sisters. Despite scaling and root planing, the periodontal lesions still progressed, but the PMN functions evaluated were now normal in both sisters. An orally asymptomatic but dermatologically affected brother showed no significant defect in the phagocytic activity and the production of superoxide radicals. PMID- 8227455 TI - Compliance perceived by Dutch periodontists and hygienists. AB - Compliance is defined here as the extent to which a patient's behavior is in accordance with the health advice he has received. On the basis of factors which could affect compliance, a quantitative survey was conducted for a group of 38 periodontists and their senior dental hygienists. The opinions of periodontists and hygienists in perceiving compliance were examined. Although there was agreement between periodontists and hygienists on some well-established aspects of compliance, the survey revealed that the role of the hygienist is of considerable consequence: hygienists appear to be more patient-oriented than periodontists. Furthermore, it is stated that poor understanding of recommendations by patients needs more attention. PMID- 8227456 TI - Effect of periodontal root planing on dentin permeability. AB - The purpose of this study was to quantitate the effects of root planing on the permeability of human root dentin in vitro. Unerupted 3rd molars were used. The crowns were removed and longitudinal slices made of the root. The hydraulic conductance of the root dentin was measured before and after root planing, acid etching and potassium oxalate application using a fluid filtration method. The results showed that root planing creates a smear layer that reduces the permeability of the underlying dentin. However, this smear layer is acid labile. Thus, root planing may ultimately cause increased dentin permeability and the associated sequelae of sensitive dentin, bacterial invasion of tubules, reduced periodontal reattachment and pulpal irritation. PMID- 8227457 TI - Relationship between palato-radicular grooves and localized periodontitis. AB - A total of 404 maxillary lateral and central incisors were examined in a group of 101 individuals comprising 64 males and 37 females for the presence of palato radicular groove (PRG). The methods used to document the presence of PRG situated at or apical to the cemento-enamel junction (CEJ) included clinical probing, flap operation and inspection with the aid of an enlarging oral mirror. PRGs were classified by location in the mesial, distal and midpalatal areas of affected teeth. The plaque index (P1I), gingival index (GI) and probing pocket depth (PPD) were recorded on the lingual aspect of the teeth examined. Those PRG with initial PPDs of > or = 4 mm and located at or apical to the CEJ were counted as PRG with PPD. The difference in the PRG location for teeth with and without PPDs was analyzed via the chi 2 test. The relationship between the presence of a PRG and the patients periodontal health and plaque accumulation were analyzed using the t test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227458 TI - The effect of sibling relationship on the periodontal condition. AB - The purpose of this investigation was to study clinically as well as microbiologically the effect of sibling relationship on the periodontal condition in a young population with a relatively high prevalence of periodontal disease and deprived from regular dental care. In this study, 23 family units consisting of 3 more siblings were evaluated. In all, 78 subjects aged 15 to 25 years were included in the study. The mean interproximal amount of loss of attachment in this population was 0.29 mm. The individual mean ranged from 0 to 1.27 mm. In 33% of the subjects, > or = 1 sites with a probing depth of 5 mm or more in conjunction with 2 mm of attachment loss were present. The results show a significant sibship effect for: plaque, calculus, loss of attachment, spirochetes on the tongue and in the pocket, Porphyromonas gingivalis on the gingiva and in the saliva and Prevotella intermedia in the saliva. These results support the hypothesis that periodontitis aggregates in families. PMID- 8227459 TI - Postmarketing surveillance: curriculum for the clinical pharmacologist. Part I: Postmarketing surveillance within the continuum of the drug approval process. AB - This series of two articles on postmarketing safety surveillance has been developed for use in training clinical pharmacologists. It provides a basic overview useful to clinical pharmacologists in a range of occupational functions. These reports can be used as the basis for a lecture or as background reading material for establishing a unit on postmarketing surveillance. For teaching rounds, multiple illustrations and summary tables have been included. These can readily be made into 35-mm slides or transparencies. Table I outlines the curriculum on postmarketing surveillance for the clinical pharmacologist. The first article describes postmarketing surveillance within the continuum of the drug approval process. The relationship of clinical trials to safety surveillance after drug approval are reviewed. The importance of spontaneous adverse drug experience reporting also is discussed. The second article focuses on the regulatory aspects of postmarketing surveillance and discusses the FDA's (Food and Drug Administration) Spontaneous Reporting System. The two types of adverse experience reports in the Spontaneous Reporting System are described: reports voluntarily submitted by health care providers and others, and reports manufacturers are required by regulation to submit. The clinical pharmacologist's role in postmarketing surveillance is defined. PMID- 8227460 TI - Pharmacokinetics of didanosine and ketoconazole after coadministration to patients seropositive for the human immunodeficiency virus. AB - The steady-state pharmacokinetics of didanosine (DDI) and ketoconazole (KET) were evaluated when the agents were administered alone or concurrently to patients seropositive for the human immunodeficiency virus. Using a randomized, three-way crossover design, multiple oral doses of DDI (375 mg twice daily for 4 days), KET (200 mg daily for 4 days) or the combination were administered under fasting conditions. When DDI and KET were coadministered, KET was given 2 hours before the morning dose of didanosine. Serial blood samples and total urine output were a collected after the administration of a final single dose on day 5 of each treatment session. Samples were analyzed using high-pressure liquid chromatography (HPLC)/ultraviolet (UV) or fluorescence methods specific for unchanged DDI (plasma and urine) or KET (plasma only). Pharmacokinetic parameters were calculated using noncompartmental methods. The average DDI maximum peak plasma concentration (Cmax) value at steady state was significantly less when DDI was administered with KET (1836 ng/mL) than when DDI was administered alone (2094 ng/mL), although the magnitude of the decrease was only 12%. Didanosine area under the curve (AUC(0-tau)) for the combination (2872 hr.ng/mL) was 8% less than when DDI was given alone (3107 hr.ng/mL); the difference was not significant. There were no significant differences among the other evaluated parameters (time to reach peak concentration [tmax], half-life [t1/2], renal clearance [CLR], or urinary recovery [UR]) between the two DDI treatments. There were no significant differences among any of the pharmacokinetic parameters between the two KET treatments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227461 TI - Comparison of vancomycin pharmacokinetics in hospitalized elderly and young patients using a Bayesian forecaster. AB - Limited data have been published that compare the pharmacokinetic parameters of vancomycin in elderly versus young patients. This study was designed to assess vancomycin pharmacokinetics in 148 elderly (> or = 60 years of age) and 140 young (18-59 years of age) hospitalized infected patients. Serum vancomycin concentrations were determined using fluorescence polarization immunoassay. Serum concentration-versus-time data were fitted to a two-compartment Bayesian forecasting program. Elderly versus young vancomycin pharmacokinetic parameters derived were as follows (patients with serum creatinine < or = 1.5 mg/dL); mean +/- standard deviation terminal disposition half-life (t1/2) of 17.8 +/- 11.8 versus 7.5 +/- 6.7 hours, respectively, P < .05; volume of distribution (Vz) of 74.2 +/- 32.3 versus 67.0 +/- 30.7 L, respectively, P = .16; and total body clearance (CL) of 0.71 +/- 0.41 versus 1.22 +/- 0.50 mL/min/kg, respectively, P < .05. Comparing subjects with normal serum creatinine values (< or = 1.5 mg/dL), the elderly required smaller daily doses as compared with the young group to maintain target peak and trough vancomycin serum concentrations (18.2 +/- 5.8 verus 25.2 +/- 7.8 mg/kg/day, P < .05). Stepwise multiple regression models for the pharmacokinetic parameters were developed to assess the predictive power of age, controlling for the effects of gender, total body weight, serum creatinine, and creatinine clearance. Age was consistently an independent and significant predictor of t1/2, Vz, and CL. These data demonstrate that elderly patients exhibit significant differences in vancomycin pharmacokinetic parameters compared with young patients and constitute a patient population in need of individualized vancomycin dosing due to substantial heterogeneity in physiologic and pharmacokinetic parameters. PMID- 8227462 TI - In vitro activity of Ro 23-9424, a dual-acting cephalosporin-quinolone antimicrobial agent. AB - In vitro activity of new dual-acting antibacterial Ro 23-9429 was tested against 1294 bacterial isolates from patients in a major tertiary care referral hospital in Saudi Arabia. Its activity was compared with that of ciprofloxacin, fleroxacin, ampicillin, cephalothin, cefoxitin, cefotaxime, ceftazidime, piperacillin, oxacillin, gentamicin, amikacin, imipenem, and vancomycin. Of the 621 members of Enterobacteriaceae tested, every single isolate was inhibited by Ro 23-9429 at minimum inhibitory concentration ranging between < .03 and 8 micrograms/mL. No other antimicrobial tested was as active as this dual-acting cephalosporin-fluoroquinolone. Similarly, all of the 255 isolates of Acinetobacter, Aeromonas hydrophila, Pseudomonas aeruginosa, and Xanthomonas maltophilia were susceptible to Ro 23-9429. It inhibited all the 120 isolates of methicillin-resistant Staphylococcus aureus. Its in vitro activity against coagulase-negative staphylococci and enterococci was superior or comparable to that of other drugs that are commonly used in clinical practice. PMID- 8227463 TI - Single-dose pharmacokinetics of cetirizine in patients with chronic liver disease. AB - The pharmacokinetics of the H1-receptor antagonist cetirizine were studied from 0 to 72 hours after a single dose of 20 mg in 5 patients with chronic hepatocellular liver disease (group A), in 5 patients with chronic cholestatic liver disease (group B), and in 16 healthy volunteers. The renal function of patients and volunteers was normal (creatinine clearance > or = 70 mL/min). Cetirizine pharmacokinetics were similar in the two groups of patients. The elimination t1/2 was prolonged in patients (mean +/- standard deviation; group A: 14.32 +/- 2.30 hours; group B: 13.86 +/- 3.14 hours) in comparison with the values observed in volunteers (9.42 +/- 2.4 hours). A reduced apparent oral body clearance also was observed in patients (group A: .48 +/- .23 mL/min/kg; group B: .41 +/- .09 mL/min/kg) in comparison with volunteers (.74 +/- .19 mL/min/kg). No differences were observed in the mean cumulative urinary excretion between patients (group A: 69 +/- 15%; group B: 69 +/- 13%) and volunteers (70.7 +/- 7.8%). PMID- 8227464 TI - Concentration-dependent protein binding of a novel oral thromboxane synthase inhibitor--FCE 22,178. AB - The protein binding of FCE 22,178 in humans was determined ex vivo by equilibrium dialysis using plasma samples obtained from a dose-ranging study in normal male volunteers. These data suggested that FCE 22,178 may exhibit concentration dependent protein binding over an in vivo concentration range of .8 to 64 micrograms/mL. Increase in free fraction at higher plasma drug concentrations corresponded directly to the dose-dependent increase in renal drug clearance. Nonlinear parameter estimation showed that FCE 22,178 binds tightly to plasma proteins with an apparent equilibrium association constant of 1.44 x 10(5) mol/L. Predicted change in the free fraction is consistent with the observed changes in renal clearance. PMID- 8227465 TI - Pharmacokinetics of cyclosporine and multiple-dose diclofenac during coadministration. AB - Pursuant to anecdotal case reports of a possible drug-drug interaction between cyclosporine and diclofenac, an open, two-period crossover study in 24 healthy male volunteers was undertaken in which a single oral dose of 300 mg cyclosporine was administered alone and on day 8 of multiple oral dosing of 50 mg diclofenac every 8 hours. Serial blood samples were obtained over 48 hours after each cyclosporine dose and over a dosing interval for diclofenac on day 7 (diclofenac alone) and day 8 (coadministration of diclofenac with cyclosporine). The mean pharmacokinetic characteristics of cyclosporine were unchanged during coadministration with diclofenac. Based on area under the curve comparison, lack of a pharmacokinetic interaction was conclusively demonstrated for the extent of cyclosporine absorption. The diclofenac maximum plasma concentration and area under the curve over a dosing interval were significantly increased during coadministration; however, a straightforward interpretation of the statistical results was confounded by pronounced variability in diclofenac pharmacokinetics. The results underscore the need for continued caution when cyclosporine and diclofenac are coadministered. PMID- 8227466 TI - Clinical evaluation of H1-receptor and H2-receptor antagonists for acute postoperative pain. AB - The acute analgesic activity of an H1-histamine antagonist, terfenadine 60 mg, and an H2-histamine antagonist, ranitidine 150 mg, were compared with ibuprofen 600 mg and placebo in a double-blind, placebo-controlled, parallel-group study. Treatments were administered to a total of 127 patients 1 hour before oral surgery. Analgesia was assessed every 30 minutes for 240 minutes after surgery. Analgesic efficacy was compared using the following standard pain intensity scales: visual analog scale, category, graphic rating, and global evaluation. Ibuprofen was significantly better than all other treatments for all measures of analgesic activity. The effects of terfenadine and ranitidine were similar to placebo. These data indicate that pretreatments with a single dose of a histamine receptor antagonist specific for either the H1- or H2-receptor does not produce analgesia in an oral surgery model of acute pain with overall assay sensitivity, suggesting that antihistamines that act primarily at peripheral sites are devoid of analgesic activity. These data contrast with other studies that have demonstrated analgesia using centrally acting antihistamines such as hydroxyzine, phenyltoloxamine, or orphenadrine. PMID- 8227467 TI - Acadesine (AICA-riboside): disposition and metabolism of an adenosine-regulating agent. AB - Acadesine (AICA-riboside) is a purine nucleoside analog with anti-ischemic properties that is currently being studied (Phase 3) for the prevention of adverse cardiovascular outcomes in patients undergoing coronary artery bypass graft (CABG) surgery. The safety, tolerance, and pharmacokinetics of the drug have previously been reported in this journal (J Clin Pharmacol 1991; 31:342 347). Recently, the authors studied the disposition and metabolism of acadesine in healthy males (n = 4) after a 15-minute intravenous infusion of 25 mg/kg of 2 14C-acadesine. The postinfusion total 14C concentrations in plasma declined in a multiexponential manner, and the terminal phase had an apparent t1/2 of about 1 week. Intact acadesine was only measurable for 2 hours after infusion. Total plasma clearance was 2.2 +/- 0.2 L/hour/kg, the acadesine blood/plasma ratio was unity, and plasma protein binding was negligible (approximately 1%). Uric acid, the end product of purine metabolism in humans, was the major metabolite of acadesine in plasma and accounted for all of the total, plasma 14C at 6 hours after infusion. In whole blood, acadesine 5'-monophosphate was present in the red blood cells, and the nucleotide represented 30% of the total blood 14C at the end of the infusion. The nucleotide was confined to the RBCs and was not present in plasma. Urine and fecal recoveries over 2 weeks accounted for 48% of the total 14C dose, with 44% excreted in urine and 4% in feces. Only 5% of the dose was excreted in urine as intact acadesine. Uric acid was the major metabolite in urine together with small amounts of hypoxanthine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227468 TI - Selectivity of acebutolol, atenolol, and metoprolol in healthy volunteers estimated by the extent the drugs occupy beta 2-receptors in the circulating plasma. AB - The selectivity of acebutolol, atenolol, and metoprolol in healthy volunteers was estimated by determining the extent to which the drugs occupied beta 1-receptors of rabbit lung and beta 2-receptors of rat reticulocytes in the circulating plasma after drug intake. This ex vivo method had the advantage of including all drug components contributing to the drug-receptor equilibrium in vivo and of excluding the factors regulating organ sensitivity to catecholamine stimulation. The oral doses of 400 mg acebutolol, 100 mg atenolol, and 100 mg metoprolol were administered to six healthy male volunteers using a double-blind, randomized, and cross-over study design. The three drugs occupied beta 1-receptors to a similar extent at 2 hours after drug intake. The receptor fraction occupied by metoprolol at 3 to 8 hours after drug intake was usually smaller, however (analysis of variance for repeated measures, P < .05) than that of the other drugs. Acebutolol occupied significantly larger fractions of beta 2-receptors (analysis of variance for repeated measures, P < .05) than did atenolol and metoprolol. Therefore, at an identical beta 1-receptor occupancy, the beta 2-receptor occupancy of acebutolol was larger than that of the other agents. Apparently, active metabolites decreased markedly the selectivity of acebutolol, but not that of metoprolol. The receptor occupancy of the agents was well in agreement with the literature concerning the selectivity, intensity, and time-course of drug actions after identical doses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227469 TI - Assessment of blood pressure during treatment with naproxen or ibuprofen in hypertensive patients treated with hydrochlorothiazide. AB - This study determined the effect of nonsteroidal anti-inflammatory drug (NSAID) administration on blood pressure in hypertensive patients taking hydrochlorothiazide (HCTZ). Ninety-seven patients with mild essential hypertension and a musculoskeletal indication for NSAID use were studied in a three-phase, multi-center, double-blind, randomized, parallel study based in 15 academic and community clinics. Patients served as their own controls. Patients with stable hypertension, not taking antihypertensive or NSAID medications, were treated with HCTZ 50 mg/day. After 4 to 5 weeks of treatment and documented stable blood pressure, naproxen 375 mg twice a day or ibuprofen 800 mg three times a day was added. Blood pressure was measured at 2 and 4 weeks of NSAID therapy. The average diastolic blood pressure was 97.5 +/- 2.4 mm Hg and the average of the mean arterial pressure (MAP) was 116.8 +/- 6.04 before treatment with HCTZ. Hydrochlorothiazide treatment decreased diastolic blood pressure to 83.1 +/- 5.6 mm Hg, and MAP to 101.1 +/- 6.5 mm Hg. With naproxen or ibuprofen treatments, mean diastolic blood pressure increased less than 3 mm Hg. At 2 weeks, ibuprofen increased diastolic blood pressure by 2.6 mm Hg (P = .004) and naproxen increased diastolic blood pressure 0.7 mm Hg (P = .40). Both ibuprofen and naproxen significantly increased diastolic pressure at 4 weeks (2.1 mm Hg, P = .042; and 1.8 mm Hg, P = .043, respectively). There was no correlation between the pre-NSAID blood pressure and the magnitude of change after 2 or 4 weeks of treatment. Changes in MAP reflected a pattern similar to diastolic pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227470 TI - Labetalol in hypertension during the third trimester of pregnancy: its antihypertensive effect and pharmacokinetic-dynamic analysis. AB - The hypotensive effect, kinetics, and concentration-response relationship of labetalol, alpha beta- and alpha 1-adrenoceptor blocking drug, were studied in seven women with a moderate-to-severe hypertension (averaged diastolic blood pressure [DBP] of 100 to 120 mm Hg measured during a 1- to 2-day hospitalization period) during the third trimester of pregnancy who received the oral twice-daily doses of 150 to 450 mg. These dosages were individually selected by attaining a therapeutic goal of DBP < or = 100 mm Hg or systolic blood pressure (SBP)/DBP reduction of > 30/15 mm Hg, as compared with the pretreatment value, at any time during the 12-hour dosing interval for a 3- to 5-day dosage escalation period. Labetalol concentrations in plasma were measured by a high-performance liquid chromatography with fluorescence detection, and the plasma drug concentration response relationship was analyzed by a sigmoidal Emax model. Labetalol decreased significantly (P < 0.05 to 0.01) the pretreatment SBP/DBP (166.3 +/- 5.2/110.3 +/ 3.0 mm Hg, mean +/- SEM) without any recognizable side-effects during the twice daily dosing period in the mothers. Peaked concentrations occurred at 1 hour postdose in all patients. The elimination half-lives ranged from 4.3 to 6.9 hours, and the apparent oral clearance from 31.9 to 73.3 mL/min/kg. The pharmacodynamic parameters (Emax and EC50) analyzed by the Emax model revealed a 3- to 5-fold interindividual variability. The gestational ages at delivery ranged from 34 to 37 weeks, and the birth weights were < 2000 g in 6 of the 7 neonates. Four neonates developed respiratory distress syndrome after delivery, and one infant died of pulmonary hypoplasia 3 months later. The results indicate that 1) labetalol orally administered in a twice-daily regimen as done in this study is an effective antihypertensive drug in women with hypertension during late pregnancy, and 2) interindividual variability in the kinetic factor (e.g., oral clearance) as well as that in the pharmacodynamic factor (e.g., EC50) appear to be related to the overall variability in the hypotensive responsiveness to the drug. However, whether labetalol and/or hypertension per se would have been related to the fetal outcome remains unanswered from the present study. PMID- 8227471 TI - Integrating worldwide marketing needs and clinical research. AB - In this era of globalization, expanding research needs, and the necessity of sharing global technologies and worldwide data evaluation techniques, a challenging environment has been created for multinational drug development, clinical testing, and marketing. Integrating worldwide marketing needs and clinical research will become even more significant in the future as harmonization of pharmaceutical industry research and regulatory requirements increase with the unification of the European Economic Community. Marketing and research teams within a pharmaceutical company must work closely together to make the drug being developed appropriate to a global market. PMID- 8227472 TI - Branded generics. PMID- 8227473 TI - Drug therapy of the fetus. AB - The fetus has become an intended object of drug therapy administered through the mother with the successful prevention of hyaline membrane disease with glucocorticoids. Maternal drug treatment has now been undertaken for a variety of fetal problems, including arrhythmias, congestive heart failure, infections, and inborn errors of adrenal metabolism. Interestingly, this planned maternal drug exposure during pregnancy coincided with increasing concerns during the last two decades about inadvertent transplacental exposure of the fetus to licit and illicit drugs. Efforts to direct drug therapy to the fetus have pointed out important gaps in knowledge of the pharmacology of the maternal-placental-fetal unit (MPFU), whereas other observations illustrated recognized principles of the pharmacology of the MPFU. Many of these principles fit the basic framework of pharmacokinetics: absorption, distribution, metabolism and excretion. Rapid changes in maternal-placental physiology and fetal development during gestation, however, lead to dramatic variations in these processes throughout pregnancy. PMID- 8227474 TI - A one-month drug analysis laboratory rotation for doctor of pharmacy students. AB - A one-month drug analysis laboratory rotation for PharmD students is described. The students gain hands-on experience through the performance of a structured sequence of laboratory experiments covering topics such as calibration standards, the calibration curve, extraction recovery, chromatographic resolution, HPLC detectors, and plasma protein binding. The self-paced, independent study approach to the rotation was cited by students to be an effective method for teaching a large number of new concepts in a short time. The course was a good aid in helping students to decide if they wanted to pursue postdoctoral research training. PMID- 8227475 TI - Carryover bias in clinical investigations. AB - If the effect of a treatment continues after the treatment is withdrawn then the response to a second treatment may well be due in part to the previous treatment. This, so called, carryover effect may bias any type of study in which subjects are tested more than once. Crossover studies can be routinely checked for this bias. In other study designs, however, common sense and alertness for unusual patterns in the data are the only defenses against it. The amount of carryover bias can be somewhat minimized by the following measures. Dose-response studies, dose-titration studies, and open-evaluation studies should require a sufficient washout period between the administrations of drugs. The use of duplicate standard deviations for the estimation of intraindividual reproducibility of a test should always be combined with a statistical test for differences between the duplicate data. Subjective variables, which are frequently influenced by psychologic carryover effects, should be validated together with objective variables whenever possible. In spite of these measures, many cases of carryover effect remain unpreventable. We shall simply have to accept them. PMID- 8227476 TI - Pharmacokinetics of methylprednisolone hemisuccinate and methylprednisolone in chronic liver disease. AB - The disposition of methylprednisolone (MP) and its prodrug hemisuccinate (MPHS) was assessed in six middle-aged patients with chronic liver disease (CLD) and compared with six younger, healthy subjects after a single IV dose of 25.4 mg of MPHS. Blood and urine samples were collected over 12 hours. Plasma and urine concentrations of MPHS and MP and plasma cortisol were measured by HPLC. MPHS clearance (CL) was significantly reduced in the CLD group (495 vs. 1389 mL/hr/kg) whereas volume of distribution (Vss) of MPHS (about 0.35 1/kg) did not differ. The elimination half-life, t1/2 beta, was significantly longer in CLD (0.61 vs. 0.32 hr). The percent recovery of unchanged MPHS in urine was similar (about 9%) in both groups. The kinetic parameters of MP did not differ between the two groups for: clearance (about 370 L/hr/kg IBW), Vss (about 1.3 L/kg), and t1/2 beta (about 3.0 hr). The suppression t1/2 of cortisol after MPHS was longer (3.9 vs. 1.9 hr) indicating metabolic pathways for cortisol and MP are affected differently in CLD. Reduction in MPHS CL may reflect altered hepatic blood flow due to both cirrhosis and age effects. However, good availability of MP from MPHS and lack of perturbation of MP pharmacokinetics in CLD patients may provide therapeutic advantages in selection of this glucocorticoid. This is the first study that characterizes the disposition of the prodrug MPHS and the formation of MP simultaneously in CLD patients. PMID- 8227477 TI - Codeine disposition in sickle cell patients compared with healthy volunteers. AB - The pharmacokinetics of codeine were determined after oral administration of codeine sulfate (60 mg) with sickle cell patients (SCPs) and healthy controls (HCs). Plasma concentrations of codeine were measured by reversed-phase high pressure liquid chromatography with fluorescence detection. Pharmacokinetic parameters were calculated using both compartmental and noncompartmental analysis. No significant differences were observed in time to reach maximum peak plasma concentration (tmax) (1.0 +/- 0.4 versus 1.4 +/- 1.0 hours), maximum peak plasma concentration (Cmax) (172 +/- 25 versus 225 +/- 97 ng/mL), area under the curve (AUC infinity) (590 +/- 96 versus 779 +/- 234 ng*h/mL), and Cl/F (104 +/- 17 versus 89 +/- 27 L/h) between SCPs and HCs. Conversely, significant differences were observed in mean residence time (MRT) (3.7 +/- 0.3 versus 4.7 +/ 0.3 hours) and half-life (t1/2) (1.7 +/- 0.2 versus 2.8 +/- 0.3 hours). In a separate study, significant differences were observed in the in vitro plasma protein binding of codeine in SCPs (66.0 +/- 8.6%) and HCs (30.5 +/- 2.7%) as well as in vivo binding (68.4 +/- 11.1% for SCPs versus 29.2 +/- 3.4% for HCs). Codeine is a relatively high-extraction drug that is primarily eliminated by metabolism in the liver. Generally, the clearance of such drugs is approximately equal to hepatic blood flow and is not affected by changes in protein binding. Therefore, the change in t1/2 observed in SCPs can be attributed to changes in volume of distribution rather than clearance. PMID- 8227478 TI - Oral methylprednisolone acetate (Medrol Tablets) for seasonal rhinitis: examination of dose and symptom response. AB - The authors compared the effect of several doses an oral corticosteroid on symptom profile and severity in ragweed hay fever. Thirty-one patients were randomized to receive 0, 6, 12, or 24 mg methylprednisolone (Medrol Tablets [MP], Upjohn, Kalamazoo, MI). A baseline week in which no treatment was given preceded the treatment comparison. At the end of this week, symptom diaries showed that most of the subjects were experiencing moderate or severe symptoms. The corticoid produced dose-related reduction in all symptoms. The difference between placebo and 24 mg MP was significant for all the symptoms monitored, except itching, which benefited marginally. With 6 mg MP, congestion, drainage, and eye symptoms showed significant drug-placebo differences but itching, running/blowing, and sneezing did not. Not all rhinitis symptoms responded equally to corticoid treatment. Those that responded least could reflect histamine effect, which was not effectively suppressed by low-dose, short-term corticoid treatment. PMID- 8227479 TI - Effects of pyrazinoylguanidine on the glucose-fatty acid cycle in normal subjects and patients with non-insulin-dependent diabetes mellitus. AB - Pyrazinoylguanidine (PZG) reduced the hyperglycemia, hyperinsulinemia, and hyperlipidemia of patients with non-insulin-dependent diabetes mellitus (NIDDM) as well as of normal subjects receiving hydrochlorothiazide (HCTZ). Mechanisms are proposed by which PZG downregulated the elevated glucose-fatty acid cycle toward a more normal level in NIDDM patients and in non-diabetic subjects maintained on HCTZ. Despite maintenance of these NIDDM patients on their current antihypertensive medication, PZG reduced further their systolic and diastolic pressures. PZG was well tolerated by both normal and NIDDM patients. PMID- 8227480 TI - Once-daily trandolapril compared with the twice-daily formulation in the treatment of mild to moderate essential hypertension: assessment by conventional and ambulatory blood pressures. AB - A double-blind, crossover study was carried out to compare the antihypertensive efficacy of the long-acting ACE inhibitor trandolapril 1 mg administered once daily and 0.5 mg twice daily in 31 patients with mild to moderate essential hypertension. After randomization, patients entered a single-blind placebo period of 4 weeks. After a double-blind treatment of 4 weeks with either of the dosage regimens, patients were then crossed over to the alternate regimen for the last 4 weeks of the study. Conventional BP and heart rate were measured on each visit and ambulatory BP monitoring was done at baseline and at the end of each treatment phase. Conventional as well as 24-hour and awake ambulatory systolic and diastolic BPs were significantly (P < 0.001) and almost identically decreased by both once- and twice-daily formulations. Moreover, the clinical response rates (reduction in seated diastolic BP > or = 10% or diastolic BP < or = 90 mm Hg) were similar with both treatment regimens (42% vs. 45% with the once- and twice daily formulations, respectively). However, trandolapril twice daily exerted a significantly (P = 0.03) greater antihypertensive effect on systolic BP during sleep as compared with the once-daily formulation. Due to the fact that the minimal effective dose was used in this trial, further studies with higher doses should demonstrate effective 24-hour control of BP as described with other long acting ACE inhibitors. In addition, our results suggest that ambulatory BP measurements should be done in dose-response studies. PMID- 8227481 TI - The effect of phenobarbital on autonomic function and epileptogenic activity induced by the hippocampal injection of penicillin in cats. AB - This study addressed whether penicillin-induced epileptiform discharges in the right hippocampus produced associated autonomic dysfunction. The study also examined the effect of phenobarbital on the heart rate and blood pressure changes that were induced by the epileptiform discharges. The delay in onset of epileptiform activity at the site of injection ranged from 1 second to 16 minutes, and consisted of interictal discharges or ictal discharges. With the onset of epileptiform activity, blood pressure and heart rate increased significantly from control (P < .05). Electrocardiogram alterations included: P-R interval changes; increased P-wave amplitude; QRS complex changes; T-wave inversion; and ST elevation. Phenobarbital 20 mg/kg intravenously suppressed the epileptogenic activity and depressed the blood pressure and heart rate below control (P < .05). In an additional series of experiments, penicillin G injected into the right hippocampus in five cats produced epileptiform activity and increased the blood pressure and the heart rate significantly from the control (P < .05). Phenobarbital (20 mg/kg, intravenously, and 40 mg/kg, intravenously) also prevented the penicillin-induced epileptiform activity. Phenobarbital (40 mg/kg, intravenously) reversed the effect of penicillin on the blood pressure and heart rate, to levels significantly below that of control (P < .05). Phenobarbital diminished both epileptiform activity and autonomic dysfunction. The autonomic dysfunction related to epileptiform activity induced by focal hippocampal administration of penicillin was similar to that induced by the intravenous administration of pentylenetetrazol. PMID- 8227482 TI - Single-dose safety and pharmacokinetics of a potential cognition-enhancing compound, CL 275,838, in healthy volunteers. AB - The pharmacokinetics and safety of CL 275,838, a potential cognition-enhancing compound, were studied after single escalating oral doses first in young healthy male volunteers and then in old (60-74 years) and very old (over 75 years) volunteers of both sexes. In all age groups absorption of CL 275,838 was rapid as assessed by the mean time to reach maximum plasma concentrations (Cmax) which averaged 1-2 hr, regardless of the dose administered. In young male volunteers both Cmax and area under the curve (AUC) increased proportionally with dose from 10 to 100 mg. Mean elimination half-lives (t1/2) of the parent compound (18-21 hr) and of its circulating metabolites II (20-22 hr) and IV (27-30 hr) were well comparable for the doses tested (50 and 100 mg). Age did not appreciably affect plasma Cmax of CL 275,838 or its two metabolites. Mean AUC and elimination half life did not appreciably differ between old and very old subjects given 50 mg CL 275,838, with the limitations dictated by the small number of elderly subjects examined. Compared with younger volunteers receiving comparable doses, however, the elderly had higher mean plasma AUC of the unchanged compound and its two metabolites, although the parameter varied widely between subjects. The mean elimination t1/2 (+/- SD) was longer in the elderly (38.8 +/- 19.6, 50.5 +/- 24.5 and 41.7 +/- 12.1 hr, respectively, for the parent compound and its metabolites II and IV) than in the young subjects. The cause(s) of these variations and the possible clinical implications remain to be established.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227483 TI - In vivo determinations of carbamazepine and carbamazepine-10, 11-epoxide binding parameters to serum proteins in monotherapy patients. AB - The in vivo serum protein binding parameters of carbamazepine (CBZ) and carbamazepine-10, 11-epoxide (CBZ-E), which was the main metabolite of CBZ in plasma, were determined in sera from 27 patients on CBZ monotherapy. Based on the results by recent studies, the authors assumed that CBZ and CBZ-E binding to serum proteins were composed of specific binding sites on alpha 1-acid glycoprotein (AAG) and albumin. Therefore, the authors determined the binding parameters of each compound by specific binding equation for two proteins. Association constants for drug-AAG binding were .071 L/mumol for CBZ and .016 L/mumol for CBZ-E. Conversely, those for drug-albumin binding were .00052 L/mumol for CBZ and .00072 L/mumol for CBZ-E. Within the investigated total concentration ranges in each compound, the AAG binding contributes largely to the drug-serum protein interactions. Furthermore, our results indicate that the albumin binding contributes to the nonsaturable serum protein binding of these compounds in the therapeutic range. PMID- 8227484 TI - Treatment of stable chronic bronchitis with iodinated glycerol: a double-blind, placebo-controlled trial. AB - In a double-blind trial, 29 patients were treated for stable, chronic bronchitis with iodinated glycerol (Organidin, 60 mg four times daily; n = 16) or placebo (n = 13). The study consisted of a 2-week single-blind lead-in period, followed by a 6-week double-blind treatment period, and assessed a total of 50 response variables. In this set of patients, treatment with iodinated glycerol was associated with statistically significant (P < 0.05) and/or positive trend (P < 0.1) toward improvement in cough frequency, cough severity, difficulty in raising sputum, sputum thickness, sputum stickiness, forced expiratory flow at FVC25% 75%, cell concentration in sputum, and several measures of physician assessment of patient condition. Iodinated glycerol was well tolerated during the 6-week treatment period; no adverse effects were reported and no patient receiving iodinated glycerol withdrew from the study prematurely. In conclusion, this study demonstrates that iodinated glycerol, 60-mg tablets given in a total daily dose of 240 mg, is safe and effective therapy for reducing symptoms in patients with stable, chronic bronchitis. PMID- 8227485 TI - Steady-state pharmacokinetics of theophylline in COPD patients treated with dirithromycin. AB - Steady-state theophylline pharmacokinetic parameters were studied in a panel of 14 patients with chronic obstructive pulmonary disease (COPD). Pharmacokinetic parameters were evaluated before, during, and after a 10-day regimen of the macrolide antibiotic, dirithromycin. The addition of dirithromycin (500 mg orally once daily at 7:00 AM) to a sustained-release theophylline dosing regimen (every 12 hours) elicited small changes in the steady-state pharmacokinetics of theophylline, which were not statistically significant. Mean steady-state plasma theophylline trough concentrations (Css,min) were invariant before, during, and after dirithromycin treatment. Mean average steady-state plasma theophylline concentrations (Cav) declined by 7% during dirithromycin treatment (NS), and mean peak plasma concentrations (Css,max) declined by 12% (NS). Theophylline clearance (CL/F) also remained relatively unchanged during dirithromycin treatment exhibiting an increase of only 11% (NS). Dirithromycin treatment does not significantly affect the steady-state pharmacokinetics of theophylline, and its use in COPD patients is not likely to modify treatment outcomes with theophylline. PMID- 8227486 TI - Hypertension and its treatment: perspective into the '90s. PMID- 8227487 TI - MedWatch, the new FDA adverse effects reporting system. PMID- 8227488 TI - Fluoxetine and desipramine in major depressive disorder. AB - The efficacy and safety of fluoxetine and desipramine were compared in a 6-week double-blind, parallel group study of patients with major depression. Twenty-five were studied while hospitalized for treatment, and 33 were studied as outpatients. Improvement on the Hamilton Rating Scale for Depression was significant for both treatments from week 1 through the end of the study and did not differ between the two treatments at any week. Overall, 64% of fluoxetine treated patients and 68% of desipramine-treated patients had at least a 50% reduction in Hamilton Depression score. We assessed whether improvement relatively early in treatment was predictive of categorical response at 6 weeks. Among fluoxetine-treated patients, but not desipramine-treated patients, the week 3 change in the Hamilton Depression mood item was significantly predictive of the response at 6 weeks. Patients treated with fluoxetine had significantly fewer side effects than those treated with desipramine. Desipramine, but not fluoxetine, caused a persistent increase in heart rate. The results suggest that early signs of response to fluoxetine are not dependent on achieving steady-state levels of the drug. PMID- 8227489 TI - Possible monoamine oxidase inhibitor-serotonin uptake inhibitor interaction: fluoxetine clinical data and preclinical findings. AB - The primary objective of this article is to review information pertinent to and emphasize the seriousness of a potential adverse monoamine oxidase inhibitor (MAOI)-serotonin uptake inhibitor interaction by considering, within the context of preclinical data, clinical cases in which an MAOI and fluoxetine hydrochloride, a specific serotonin uptake inhibitor, were administered in close temporal proximity. Clinical cases were identified by a review of spontaneous adverse event reports submitted voluntarily to Eli Lilly and Company through its drug surveillance program and by a review of reports of MAOI interactions published in the scientific literature. A discussion of eight selected clinical cases of acute adverse reactions (seven with fatal outcomes; one with a favorable response after cyproheptadine therapy) reported to Eli Lilly and Company in which an MAOI was initiated concurrently or shortly after the discontinuation of fluoxetine and a review of preclinical data suggest a possible role of serotonin and/or serotonin-dopamine interactions in the hypermetabolic state that may occur when a serotonin uptake inhibitor and an MAOI are coadministered, although alternative etiologic processes are possible as well. Data reviewed reinforce the idea that the administration of an MAOI in close temporal proximity to fluoxetine is contraindicated. PMID- 8227490 TI - Effect of fluvoxamine on panic disorder. AB - Several reports suggest that selective serotonin reuptake blockers are helpful in the treatment of panic disorder. The aim of the study was to compare fluvoxamine with placebo in 50 panic disorder patients by using an 8-week, double-blind, parallel-groups design. Weekly assessment included a panic attack diary (frequency and severity), the Montgomery-Asberg Depression Scale, the Clinical Anxiety Scale, and the Sheehan Disability Scale. Although both groups improved on all measures, the fluvoxamine group experienced significantly less frequent major panic attacks from the third week on and significantly lower ratings on anxiety, depression, and disability from the sixth week on. Mean ratings of the severity of major and the severity and frequency of minor attacks were not affected differently by fluvoxamine and placebo. At the end of the study, significantly more patients on fluvoxamine were free of major and minor panic attacks. The results indicate that: (1) the administration of fluvoxamine, as compared with placebo, led to a significant reduction in the number of panic attacks. (2) The severity of panic attacks was not affected by fluvoxamine. (3) The effect of fluvoxamine on anxiety, depressive mood, and disability differed from placebo only after 6 weeks of treatment, after which the placebo group showed either no further improvement or a reversal of symptoms. (4) Participation in a drug study, even without additional psychotherapy, led to significant improvement in all patients. PMID- 8227491 TI - A comparison of symptom determinants of patient and clinician global ratings in patients with panic disorder and depression. AB - The Clinical Global Impression (CGI) is a standard assessment tool that generally shows good sensitivity to change in psychopharmacology trials. However, systematic assessment has not been conducted to determine how rating decisions are made. In this article, we examine the relationship between syndromal symptomatology and the CGI severity and improvement ratings in a study of 116 patients who met DSM-III-R criteria for both Panic Disorder and Depression. Anticipatory anxiety and depression ratings were significantly associated with each CGI item. Frequency of panic attacks was consistently related to the clinician's rating of severity but was only sporadically related to the clinician and patient improvement ratings. These findings are fairly consistent during the course of treatment. Our empirical examination of symptom determinants of the CGI demonstrates that it appears to be used systematically, yet global ratings are not merely a composite of symptomatology. Its widespread application in clinical trials is well justified. PMID- 8227492 TI - Pharmacotherapy of personality disorders: conceptual framework and clinical strategies. AB - This article delineates the conceptual models used when medications are prescribed for patients with personality disorders and reviews the data on the efficacy of these medications. Studies before 1980 are difficult to interpret because of changes in diagnostic criteria. Nonetheless, early studies on non-DSM III disorders such as pseudoneurotic schizophrenia, emotionally unstable character disorder, hysteroid dysphoria, and subaffective disorders indicated the potential utility of pharmacotherapy for treating personality disorders. Models to consider in evaluating the possible use of medications for treating personality disorders are: (1) treating the disorder itself; (2) treating symptom clusters within and across disorders; and (3) treating associated axis I disorders. Among the current personality disorders, borderline personality disorder has been the most extensively studied, with antipsychotic agents being the most well-documented treatment. Monoamine oxidase inhibitors, fluoxetine, and carbamazepine show promise. Schizotypal disorders may respond to low-dose antipsychotic drugs. Although heuristically valuable, the symptom cluster approach to treatment has not yet been validated. Axis I disorders, especially depression, are frequently associated with all personality disorders. Dependent personality disorder is linked to panic disorder with agoraphobia, whereas avoidant personality disorder is associated with social phobia and panic. In general, pharmacotherapy for axis I disorders is less effective in the presence of a comorbid personality disorder. Despite the modest benefits seen in many studies, pharmacotherapy can add significantly to the overall treatment of those with personality disorders. Future research must carefully assess the effect of comorbid axis I disorders on responses. The symptom cluster/psychobiologic dimension approach should be investigated in clinical studies. PMID- 8227493 TI - The efficacy of fluvoxamine in obsessive-compulsive disorder: effects of comorbid chronic tic disorder. AB - This retrospective case-controlled analysis evaluated treatment response to the serotonin reuptake inhibitor fluvoxamine in patients with obsessive-compulsive disorder (OCD), with or without a comorbid chronic tic disorder. Thirty-three fluvoxamine-treated OCD patients with a concurrent chronic tic disorder were compared with 33 age- and sex-matched OCD patients without chronic tics who had received fluvoxamine treatment in the same setting during the same period of time and in a similar manner. Although both groups of patients demonstrated statistically significant reductions in obsessive-compulsive, depressive, and anxiety symptoms with fluvoxamine treatment, the frequency and magnitude of response of obsessive-compulsive symptoms was significantly different between the two groups. A clinically meaningful improvement in obsessive-compulsive symptoms occurred in only 21% of OCD patients with comorbid chronic tics compared with a 52% response rate in OCD patients without chronic tics. Moreover, OCD patients with a concurrent chronic tic disorder showed only a 17% reduction in Yale-Brown Obsessive-Compulsive Scale scores compared with a 32% decrease in the severity of obsessive-compulsive symptoms in those OCD patients without chronic tics. These results suggest that serotonin reuptake inhibitor monotherapy may be less efficacious for improving obsessive-compulsive symptoms in OCD patients with than without tics. Combined with differences in the clinical phenomenology between these two groups, these treatment response data support the hypothesis that OCD patients with a comorbid chronic tic disorder may be a clinically meaningful subtype. PMID- 8227494 TI - Tolerance to fluoxetine. PMID- 8227495 TI - Upjohn answers Consumer Reports. PMID- 8227496 TI - Fluoxetine responsivity mediated by serum levels, not dose. PMID- 8227497 TI - The rise of serum aminotransferases in a patient treated with bupropion. PMID- 8227498 TI - Dyskinesia associated with fluvoxamine. PMID- 8227499 TI - P450IIIA (CYP3A) activity in severely depressed patients. PMID- 8227500 TI - I have read about a patient in whom fluoxetine use may have caused lung damage. Can you comment on the likelihood that there was a cause and effect relationship in this published case? PMID- 8227501 TI - The perception and production of rhyme in normal and developmentally apraxic children. AB - The phonological competence of four children, aged 5-7 years old, who demonstrated a cluster of symptoms consistent with a diagnosis of developmental apraxia of speech (DAS), was contrasted to that of four normal children. Four rhyming tasks were used to assess the ability of the children to both spontaneously generate rhyming words to targets and to judge the (in)appropriateness of a rhyme in both a word series and forced-choice word pairs. The DAS children revealed a severe deficit in rhyming ability across all tasks and had rhyming abilities markedly inferior to those shown by normal children. The rhyming results were interpreted as possibly indicative of an impoverishment of an internalized phonemic representation system, which precludes accessing and evoking the needed sound image for the vowel + coda for a rhyme. These results, while preliminary in nature, lend support to a conceptualization of DAS as a fundamental disorder of the segmental phonological level of language that impacts on all hierarchically relevant language components. The hypothesis that the underlying etiology of DAS is a developmental dysmorphology of the neural substrates that mediate such basic phonological representational structure is discussed. PMID- 8227502 TI - The emerging phonological system of an autistic child. AB - This case report provides a detailed phonological investigation of the speech of an 8-year-old autistic boy. Three approaches were used for elicitation of speech: delayed imitation, object naming, and a connected speech sample. Phonetic inventory analysis revealed that stops, nasals, and glides were generally present, whereas fricatives, affricates, and the liquid /r/ were absent. There were also positional restrictions on the use of specific sounds. This information, together with a phonological process analysis, revealed: (a) the existence of several phonological processes that are common in normal development; (b) the persistence of several phonological processes, e.g., velar fronting, beyond the expected age; (c) the occurrence of some unusual sound changes, e.g., extensive glottal replacement and segment coalescence; (d) evidence of "chronological mismatch" (Grunwell, 1981); and (e) restricted use of contrasts (Ingram, 1976). The subject's use of phonological processes resulted in extensive homonymy, which, together with process interactions and the use of jargon, resulted in severely reduced intelligibility. This child appeared to be acquiring his phonological system in at least a partly unique way, showing some typical patterns as well as some patterns that rarely appear in normally developing children. PMID- 8227503 TI - Vocal problems among aerobic instructors and aerobic participants. AB - A questionnaire to determine the characteristics of vocal problems was distributed to 75 female aerobic instructors and 75 female aerobic participants ranging in age from 20 to 40 years. Of the questionnaires returned, 50 from each group, with completed replies to all questions, were selected for inclusion in the study. The questionnaires, constructed by the investigator, included pertinent questions relative to length of teaching/participating experience, average number of classes taught/participated in weekly, number of classes taught/participated sequentially, type of aerobics, medical history and history of vocal problems and/or conditions, water intake daily and during instructing/participating, sleep habits, and smoking habits. The results indicated that aerobic instructors generally experienced more hoarseness and episodes of voice loss during and after instructing and a significantly higher prevalence of nodules. The group differences in reported unrelated episodes of hoarseness, however, were quite similar. The results suggest that aerobic instructors experience a higher prevalence of vocal problems when compared to individuals participating in aerobics. PMID- 8227504 TI - The use of different service delivery models for children with phonological disorders. AB - A group of 12 preschool children with phonological process errors was selected, and individual subjects were randomly assigned to one of two treatments that differed in relation to service delivery. Group I received a treatment that was administered exclusively by the clinician. Group II received a combination that included clinician administered treatment and parent administered instruction with the Speech Viewer system. Results indicate that both groups improved significantly, but they did not differ significantly from each other in the degree of change. Implications with respect to the service delivery options and their respective components are discussed. PMID- 8227505 TI - Do children's reactions to peers who wear visible hearing aids always tend to be negative? PMID- 8227506 TI - Hospital reported complications of laparoscopic cholecystectomy among Medicare and Medicaid patients. AB - The purpose of this study was to initiate a hospital-based case review of all laparoscopic cholecystectomies performed on Medicare and Medicaid patients in New York State in 1991 where there were one or more complications. Another purpose was to facilitate efforts by hospitals to monitor the performance of laparoscopic cholecystectomy through an educational process of data-sharing. There were 2,940 Medicare and 1,108 Medicaid cholecystectomies in New York State in 1991. Of these, 351 (11.9%) Medicare and 107 (9.7%) Medicaid patients were reported as having complications. The complication rate for Medicare patients was slightly lower than that observed (15.8%) in an epidemiologic study of Medicare patients in New York State who underwent laparoscopic cholecystectomy during the period January 1, 1990-June 30, 1991. Both of these observed rates for Medicare patients are higher than the mean 6.0% complication rate reported for open cholecystectomy in the literature. These increased rates may in part be due to age related risk factors present among Medicare patients. The absence of age related risk factors may also largely account for the lower laparoscopic cholecystectomy complication rate (9.7%) observed among Medicaid patients. The complication rate of 9.7% for Medicaid patients is similar to rates reported in other recent studies. The 11.9% complication rate for Medicare patients is higher than that reported in other recent studies. However, careful patient selection, the absence of age related risk factors, and greater surgical experience may account for the lower complication rates reported in some published series. Overall, the coding of complications was found to be accurate. The coding of laparoscopic cholecystectomy was found to be slightly flawed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227507 TI - Illness and injury in family day care: a seasonal survey. AB - Many mothers with children under age 6 are employed and it is not usually feasible for a parent to stay home with a child who is mildly ill. Such ill children likely remain in child day care programs. The extent to which this occurs and the management of these children in family day care was studied. Over the course of a year, 1 to 3 visits were made to 714 family day care homes in order to survey providers. With enrollments ranging between 0 and 18 children per day care home at the time of each visit, information on 3,630 "child enrollment days" was collected. The median age of children in care was 2 years. An average of 16% of all children were ill on any one day (with seasonal variation); 1% were injured. Of ill children, 82% attended day care that day, 49% had contacted a physician about that illness, and 28% were administered a medication at the day care setting. Twenty-one percent of children receiving medication in day care had no contact with a physician for that illness. These data show that mild childhood illnesses are routinely managed by child day care providers. Physicians who traditionally limit their illness-management education to parents need to recognize the health education and consultation needs of day care providers. PMID- 8227508 TI - Post-hospitalization followup appointment-keeping among the medically indigent. AB - The purpose of this study was to measure followup appointment-keeping in patients discharged from a General Medicine Inpatient Service and to identify possible predictors of compliance. Patients were interviewed on hospital admission and all charts were reviewed on discharge. A subset of patients were interviewed by telephone an average of one month after first followup appointment date. The study was conducted in an urban public teaching hospital with hospital-based and community clinics. A convenience sample of 209 patients were selected from admissions to the General Medicine Inpatient Service over a three month period. Followup appointment-keeping was recorded on all 195 patients discharged alive. Seventy-five percent of patients had no medical insurance, public or private. A compliance rate of 60% (95% confidence interval: 53% to 67%) with first followup appointment was found. Variables associated with compliance and which retained independence on multiple logistic regression analysis, followed by adjusted odds ratios (95% confidence intervals) were: no copayment requirement, odds ratio 3.2 (1.6 to 6.3), single followup appointment 2.9 (1.4 to 5.9), apartment dwelling 3.2 (1.4 to 7.3) and non-primary care clinic appointment 2.3 (1.1 to 4.8). We conclude that health-care-delivery related factors such as no copayment requirements are strongly associated with appointment-keeping in a public hospital population. PMID- 8227509 TI - Emergency shelters in Vancouver, Canada. AB - The shelter experiences, employment history, income and social service needs utilization were examined among 124 emergency shelter users in Vancouver, British Columbia. Thirty-six (30.5%) reported this as their first experience with living in a shelter; the length of stay in the present shelter ranged from one to 90 days. Reasons for shelter use included: eviction from last place of residence, loss of job, or suffering from health problems rendering one unable to work. The highest unmet service needs were finding affordable housing and finding a job. This combination of scarcity of low-cost housing, health problems, and difficulty in finding employment have led to a situation where emergency shelters have become extended places of residence, a "home" to many. PMID- 8227510 TI - New York Medical College: an early center of excellence in American medical education. AB - Originating in 1850, the New York Medical College was one of the earliest medical schools to adopt the educational reforms advocated by the American Medical Association. This college lengthened the school term, employed more professors than usual and established a separate Board of Censors to approve students for their medical degrees. In addition, the New York Medical College initiated bedside clinical teaching using a 27 bed hospital under its own control. In an era when many medical schools were little more than diploma mills, the New York Medical College created educational reforms that were not adopted by other schools until many years later. Due to a combination of causes the New York Medical College only existed from 1850 to 1864. However, the effort to reform medical education was significant and should not be overlooked. Incidentally, the New York Medical College of 1850 to 1864 was in no way related to the currently existing medical school of the same name. PMID- 8227511 TI - Immunohistochemical localization of a ligand-binding and a structural subunit of nicotinic acetylcholine receptors in the central nervous system of Drosophila melanogaster. AB - The distribution of two subunits of nicotinic acetylcholine receptors in the developing and the differentiated central nervous system of Drosophila melanogaster was studied. With subunit-specific antibodies raised against the ligand-binding alpha-like subunit ALS and the putative non-ligand-binding subunit ARD, we find both ALS-like and ARD-like immunoreactivity widely distributed in most neuropiles of the optic lobes, the protocerebrum, the deutocerebrum and the thoracic ganglion of the adult fly. With a single exception, namely in the lamina of the visual system, the antigens recognized by the two types of antibodies are colocalized. This observation is consistent with previous immunoprecipitation data indicating that the ALS and ARD proteins are integral components of the same hetero-oligomeric receptor that binds the nicotinic antagonist alpha-bungarotoxin with high affinity. During embryonic development ARD-like immunoreactivity is first detectable in approximately 10 hour old embryos. Both subunits are consistently detected in the central nervous system of the late embryo, the three larval stages, and all prepupal and pupal stages. During metamorphosis the optic stalk is transiently immunoreactive with anti-ARD, but not with anti-ALS antiserum. Although in larvae and adults, immunoreactivity with both types of antibodies is most abundant in synaptic regions, in embryos and pupae strong staining of cortical cell body layers is observed, in particular with anti-ARD antisera. As these developmental periods coincide with strong accumulation of ARD transcripts, the cell body staining may reflect newly synthesized and assembled receptors, while the functional ARD- and ALS-containing receptor may be destined for synapses. PMID- 8227512 TI - Synapses between slowly adapting lung stretch receptor afferents and inspiratory beta-neurons in the nucleus of the solitary tract of cats: a light and electron microscopic analysis. AB - Previous neuroanatomic and physiologic studies indicated that afferent fibres from slowly adapting pulmonary stretch receptors (SAR) project to the nuclei of the solitary tract and terminate on inspiratory beta-neurons. In the present study we combined electrophysiologic and morphologic approaches to verify the presumed monosynaptic connections between SARs and beta-neurons. Single identified beta-neurons and single identified SAR afferent fibres were labelled intrasomally and intraaxonally, respectively, with horseradish peroxidase (HRP) in the same anesthetized cats. Under the light microscope, we analyzed the morphology of beta-neurons and their dendritic fields and of the terminal projection pattern of fibres from SARs and identified potential synaptic connections between boutons of SAR afferent fibres and the soma and dendrites of beta-neurons. The identified tissue was then processed further for electron microscopic analysis. On average, beta-neurons had 6 primary dendrites that bifurcated 3-8 times. The dendritic trees extended 1.5 mm both rostrocaudally in the ventrolateral nucleus of the solitary tract and medially into the intermediate subnucleus. Axons of beta-neurons curved toward the midline and no collateral branches were evident over its stained length (2.5-3.4 mm). Axodendritic synaptic contacts between SAR fibres and beta-neurons were identified electron microscopically in four of six tissue samples chosen by light microscopy. In addition, we located 2 axodendritic and 2 axosomatic synaptic contacts that were not observed under light microscopic screening. The boutons of SAR fibres contained clear, round vesicles and formed asymmetrical synapses with beta-neurons. Multiple synaptic connections were found between collaterals of a single SAR and single beta-neurons, indicating a dense terminal projection of single SAR afferent fibres onto beta-neurons. These morphologic data prove monosynaptic connections between electrophysiologically identified SAR afferent fibres and beta-neurons. PMID- 8227513 TI - Thalamic projections to areas 3a, 3b, and 4 in the sensorimotor cortex of the mature and infant macaque monkey. AB - Area 3a in the macaque monkey, located in the fundus of the central sulcus, separates motor and somatosensory cortical areas 4 and 3b. The known connections of areas 4 and 3b differ substantially, as does the information which they receive, process, and transfer to other parts of the central nervous system. In this analysis the thalamic projections to each of these three cortical fields were examined and compared by using retrogradely transported fluorescent dyes (Fast Blue, Diamidino Yellow, Rhodamine and Green latex microspheres) as neuron labels. Coincident labeling of projections to 2-3 cortical sites in each monkey allowed the direct comparison of the soma distributions within the thalamic space of the different neuron populations projecting to areas 3a, 3b, and 4, as well as to boundary zones between these cortical fields. The soma distribution of thalamic neurons projecting to a small circumscribed zone (diameter = 0.5-1.0 mm) strictly within cortical area 3a (in region of hand representation) filled out a "territory" traversing the dorsal half of the cytoarchitectonically defined thalamic nucleus, VPLc (abbreviations as in Olszewski [1952] The Thalamus of the Macaca mulatta. Basel: Karger). This elongate, rather cylindrical, territory extended caudally into the anterior pulvinar nucleus, but not forward into VPLo. The rostrocaudal extent of the thalamic territory defining the soma distribution of neurons projecting to small zones of cortical area 3b was similar, but typically extended into the ventral part of VPLc, filling out a medially concavo convex laminar space. Two such territories projecting to adjacent zones of areas 3a and 3b, respectively, overlapped and shared thalamic space, but not thalamic neurons. Contrasting with the 3a and 3b thalamic territories, the soma distribution of thalamic neurons projecting to a circumscribed zone in the nearby motor cortex (area 4) did not penetrate into VPLc, but instead filled out a mediolaterally flattened territory extending from rostral VLo, VLm, VPLo to caudal and dorsal VLc, LP, and Pul.o. These territories skirted around VPLc. All three cortical areas 4, 3a, and 3b) also received input from distinctive clusters of cells in the intralaminar Cn.Md. It is inferred that, in combination, the thalamic territories enveloping those neuron somas projecting to, say, the sensorimotor hand representation in areas 3a, 3b, and 4 (and also areas 1 and 2), which would be coactive during the execution of a manual task, constituted a lamellar space extending from VLo rostrally to Pul.o caudally.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8227514 TI - Ipsilateral cortical projections to areas 3a, 3b, and 4 in the macaque monkey. AB - In the macaque monkey area 3a of the cerebral cortex separates area 4, a primary motor cortical field, from somatosensory area 3b, which has a subcortical input mainly from cutaneous mechanoreceptive neurons. That each of these cortical areas has a unique thalamic input was illustrated in the preceding paper. In the present experiments the cortical afferent projections to these 3 areas of the sensorimotor cortex monkey were visualized and compared, using 4 differentiable fluorescent dyes as axonal retrogradely transported labels. The cortical projection patterns to areas 3a, 3b, and 4 were similar in that they each consisted of (a) a "halo" of input from the immediately surrounding cortex, and (b) discrete projections from one or more remote cortical areas. However, the pattern of remote inputs from precentral, mesial, and posterior parietal cortex was different for each of the 3 cortical target areas. The cortical input configuration was least complex for area 3b, its remote input projecting mainly from insular cortex. The pattern of discrete cortical inputs to the motor area 4, however, was more complex, with projections from the cingulate motor area (24c/d), the supplementary motor area, postarcuate cortex, insular cortex, and postcentral areas 2/5. Area 3a, in addition to the proximal projections from the immediately surrounding cortex, also received input from the supplementary motor area, cingulate motor cortex, insular cortex, and areas 2/5. Thus, this pattern of cortical input to area 3a resembled more closely that of the adjacent motor rather than that of the somatosensory area 3b. Contrasting with this, however, the thalamic input to area 3a was largely from somatosensory VPLc (abbreviations from Olszewski [1952] The Thalamus of the Macaca mulatta. Basel: Karger) and not from VPLo (with input from cerebellum, and projecting to precentral motor areas). PMID- 8227515 TI - Correlations between the receptive field properties and morphology of neurons in the deep layers of the hamster's superior colliculus. AB - Extracellular and intracellular recording, receptive field mapping, and intracellular HRP injection techniques were used to define the morphological classes of cells in the deep laminae of the hamster's superior colliculus and to determine whether there are any correlations between the structural and functional characteristics of these neurons. A total of 110 neurons were characterized and reconstructed. Of these, 23.6% (N = 26) were visual, 60% (N = 66) were somatosensory, 0.9% (N = 1) were bimodal (visual-somatosensory), and 15.4% (N = 17) were unresponsive. Of the somatosensory neurons, 72.7% (N = 48) were low threshold, 4.5% (N = 3) had a wide dynamic range, 9.1% (N = 6) responded only to noxious stimulation, and 13.6% (N = 9) had complex somatosensory receptive fields. Deep layer cells were divided into eight morphological classes. These classes were multipolar cells (26.4%, N = 29), bipolar cells (9.1%, N = 10), widefield vertical cells (7.3%, N = 8), horizontal cells (13.6%, N = 15), stellate cells (10.9%, N = 12), ventrally directed cells (5.5%, N = 6), sparse radial cells (17.3%, N = 19), and small sparse radial cells (6.4%, N = 7). Four cells (3.6%) did not fit into this classification scheme. Univariate and multivariate analyses of variance of properties such as soma area, number of branch points, total dendritic length, and volume and orientation of dendritic arbor indicated that these classes were significantly different. However, chi 2 analysis and multivariate analysis of variance indicated no significant relationships between morphological class and either laminar location or receptive field type. There was a significant positive relationship between the possession of dendrites that extended into the superficial laminae and visual responsivity. PMID- 8227516 TI - Giant terminals in the dorsal octavolateralis nucleus of lampreys. AB - The dorsal octavolateralis nucleus of lampreys is a primary nucleus for electroreceptive stimuli in the medulla. In Lampetra japonica, the rostral and caudal thirds of this nucleus are exclusively occupied by giant terminals, which become evident when the primary fibers of an electrosensory nerve (recurrent branch of the anterior lateral line nerve) are labeled with horseradish peroxidase. We studied the ultrastructure of these terminals. They contain neurofilaments, mitochondria, microtubules, and tubular membranous structures. Many synapses, all of the chemical type, are located around the neck region of the terminal swellings. Many vesicular structures, which are clear, round, and uniform in size, and most of which are probably synaptic vesicles, are densely clustered in a single large mass in the neck region of the terminals. Some of the tubular structures may serve as a membrane reservoir for the large number of synaptic vesicles required in the giant terminals. PMID- 8227517 TI - Distribution of the mRNA for a metabotropic glutamate receptor (mGluR3) in the rat brain: an in situ hybridization study. AB - Distribution of the mRNA for a metabotropic glutamate receptor, mGluR3, which is coupled to the inhibitory cAMP cascade, was examined in the central nervous system of the adult albino rat by in situ hybridization. The hybridization signals of mGluR3 were detected not only on neuronal cells but also on many glial cells throughout the brain and spinal cord. In the neuronal cells, prominent expression of mGluR3 mRNA was seen in the thalamic reticular nucleus. Moderately labeled neurons were seen in the anterior olfactory nucleus, cerebral neo- and mesocortical regions, lateral amygdaloid nucleus, ventral part of the basolateral amygdaloid nucleus, dorsal endopiriform nucleus, supraoptic nucleus, superficial layers of the superior colliculus, inferior colliculus, interpeduncular nucleus, superior olivary nuclei, and Golgi cells in the cerebellar cortex. Weakly labeled neurons were observed in the striatum, nucleus accumbens, ventral pallidum, globus pallidus, entopeduncular nucleus, lateral hypothalamic area, hypothalamic paraventricular nucleus, medial habenular nucleus, anterior pretectal nucleus, Barrington's nucleus, Nucleus O, paragenual nucleus, trigeminal sensory complex, cochlear nuclei, dorsal motor nucleus of the trigeminal nerve, dorsal cap of the inferior olive, spinal dorsal horn, and lamina X of the spinal cord. The stellate cells in the cerebellar cortex, and neurons in the deep cerebellar nuclei were also labeled weakly. The granule cell layer of the dentate gyrus, as a whole, appeared to be labeled intensely, but each of the granule cells was labeled only weakly. No significant labeling was detected in the mitral and tufted cells in the olfactory bulb, hippocampal pyramidal cells, Purkinje and granule cells in the cerebellar cortex, or somatic motoneurons. The distribution of mGluR3 mRNA in particular neurons and glial cells indicates specific roles of mGluR3 in the glutamatergic system of the central nervous system. PMID- 8227518 TI - Expiratory neurons of the Botzinger Complex in the rat: a morphological study following intracellular labeling with biocytin. AB - The term "Botzinger Complex" (BOT) refers to a distinct group of neurons, located near the rostral portion of the nucleus ambiguus, which are known to play an important role in the control of respiratory movements. Previous studies conducted in cats have demonstrated that most of these neurons are active during expiration, exerting a monosynaptic inhibitory action on several subpopulations of inspiratory neurons in the medulla and spinal cord. The aim of this study was to examine morphological properties and possible synaptic targets of BOT neurons in the rat. Forty-one expiratory neurons were labeled intracellularly with biocytin; 12 were interneurons (BOT neurons) and 29 were motoneurons. The latter could not be antidromically activated following stimulation of the superior laryngeal or vagal nerves. BOT neurons showed extensive axonal arborisations in the ipsilateral medulla, with some projections to the contralateral side. Bouton like axon varicosities mainly clustered in two areas: near the parent cell bodies, and in the area corresponding to the rostral part of the ventral respiratory group (VRG). In five pairs of labeled neurons, each consisting of one BOT neuron and one inspiratory neuron in the rostral VRG, no appositions were identified at the light microscopic level between axons of BOT neurons and dendrites or cell bodies of inspiratory neurons. These results demonstrate that some features of BOT expiratory neurons in the rat are similar to those previously described in cats. The differences include their more ventral location in relation to the compact formation of nucleus ambiguus (retrofacial nucleus), and the relative paucity in the rat of neurons displaying an augmenting pattern of activity and of neurons with spinally projecting axons. In addition, we were unable to find morphological evidence for contacts between labeled BOT neurons and ipsilateral inspiratory neurons near the obex level, a finding not consistent with previous electrophysiological studies in the cat in which such synaptic connections have been identified. PMID- 8227519 TI - Vibrissal stimulation affects glucose utilization in the trigeminal/somatosensory system of normal rats and rats prenatally exposed to ethanol. AB - The effect of gestational ethanol exposure on stimulus-induced sensory activity in the trigeminal/somatosensory system was determined. The mature offspring of mothers fed an ethanol-containing diet (Et) or pair-fed a nutritionally matched control diet (Ct) were examined. The C-row mystacial whiskers were stimulated. Glucose utilization in the principal sensory nucleus of the trigeminal nerve (PSN), ventrobasal thalamus, and somatosensory cortex was determined with [14C]2 deoxyglucose autoradiography. In Ct- and Et-treated rats, whisker stimulation increased glucose utilization in C-row barrel(oid)s in the left PSN, the right ventrobasal thalamus, and the right somatosensory cortex. The rate of glucose utilization in the C-row barrel(oid)s and in nonstimulated regions was lower in the Et-treated rats than in controls. In the cortices of Ct-treated rats, the activity in the C-row barrels on the right side was greater than in the right nonbarrel somatosensory cortex. Et-treated rats also exhibited an increase in glucose utilization, albeit smaller than that in the Ct-treated rats. In contrast, the glucose utilization in the left B- and C-row barrels of Ct-treated rats was decreased. No such decrease was evident in the left cortices of Et treated rats. Thus, stroking whiskers stimulates the activity of sites in the trigeminal/somatosensory system. In cortex, the definition of these sites is emphasized by depressed activity, i.e., "surround" inhibition, in sites connected via callosal or corticocortical projections. Prenatal exposure to ethanol depresses the metabolic activity regardless of the physiological state; however, the "surround" inhibition of cortical activity is eliminated by prenatal exposure to ethanol through an exuberant projection. PMID- 8227520 TI - Localization of cholinergic neurons in the forebrain and brainstem that project to the suprachiasmatic nucleus of the hypothalamus in rat. AB - In mammals, the suprachiasmatic nucleus is responsible for the generation of most circadian rhythms and their entrainment to environmental cues. Cholinergic agents can alter circadian rhythm phase, and fibres immunoreactive for choline acetyltransferase, the biosynthetic enzyme for acetylcholine, are present in the suprachiasmatic nucleus. Since there are no cholinergic somata in the suprachiasmatic nucleus, these fibres must represent the terminals of cholinergic neurons whose cell bodies are located elsewhere in the brain. This study was aimed at locating the cholinergic neurons that project to the suprachiasmatic nucleus by retrograde and anterograde tract-tracing and immunohistochemistry for choline acetyltransferase in the rat. After injection of fluorogold, a retrograde tracer, into the suprachiasmatic nucleus, retrogradely labelled neurons that were immunopositive for choline acetyltransferase were located throughout the rostrocaudal extent of the cholinergic basal nuclear complex, with highest densities in the substantia innominata and the nucleus basalis magnocellularis. A few cells were also located in the medial septum and in the vertical and horizontal limbs of the diagonal band of Broca. In the brainstem, double-labelled neurons were located in the laterodorsal tegmental nucleus, pedunculopontine tegmental nucleus and the parabigeminal nucleus. Injections of the anterograde tracer biocytin in these three brainstem nuclei resulted in fibre labelling in the suprachiasmatic nucleus, consistent with the retrograde findings. No clearly double-labelled cells were located in the retina. These results suggest that the suprachiasmatic nucleus receives cholinergic afferents from both the basal forebrain and mesopontine tegmentum which may mediate cholinergic effects on circadian rhythms. PMID- 8227521 TI - Topographical representation of shoulder motor nuclei in the cat spinal cord as revealed by retrograde fluorochrome tracers. AB - The present investigation demonstrates the morphological relationships among the main shoulder motor nuclei within the spinal cord of the cat. The intraspinal position of these nuclei has been revealed by retrograde labelling of spinal motor neurones via their peripheral nerves supplying anatomically identified shoulder muscles. Multiple pressure injection of up to four fluorescent tracers (Bisbenzimide, Fast Blue, Fluoro-Gold, Rhodamine-b-isothiocyanate) in one experiment was used to show the longitudinal distribution and topographical relations of motor neurones projecting to muscles acting on the scapulo-humerus joint. Tracer-positive cells have been found from middle C5 to rostral Th2 in the cervical cord, forming coherent longitudinal cell clusters separated in medial and lateral projection fields in the ventral horn. The present data suggest that the anatomical organization of spinal shoulder motor neurones corresponds to the embryonic origin of their later target muscles. All medial motor nuclei project to muscles deriving from ventral embryonic origins, while those motor nuclei lying in lateral positions innervate muscles originating from dorsal muscle primordia. Therefore, the spinal topography of shoulder motor nuclei seems to be independent of both the position and the function of a given muscle in the adult animal. PMID- 8227522 TI - Coexistence of NADPH diaphorase with GABA, glycine, and acetylcholine in rat spinal cord. AB - The enzyme NADPH diaphorase is present in many spinal neurons, and is thought to correspond to nitric oxide synthase. In order to determine which types of neuron in the spinal cord contain this enzyme, we have carried out a combined enzyme histochemical and immunocytochemical study with antibodies to GABA, glycine, and choline acetyltransferase. Two hundred rats were tested for GABA- and glycine like immunoreactivity. The majority of these neurons (207/224) were GABA immunoreactive and 139 were also glycine-immunoreactive. NADPH diaphorase positive neurons in laminae I and II generally showed both types of immunoreactivity, while those in deeper laminae of the dorsal horn and around the central canal either showed both types or else were only GABA-immunoreactive. Since GABA and acetylcholine are thought to coexist in spinal neurons, NADPH diaphorase staining was combined with immunostaining for choline acetyltransferase. Immunoreactive neurons in laminae III and IV were all NADPH diaphorase-positive, while only some of those around the central canal and in the deeper laminae of the dorsal horn were positive. Choline acetyltransferase immunoreactive neurons in the intermediolateral cell column (presumed sympathetic preganglionic neurons) were often NADPH diaphorase-positive, whereas those in the ventral horn (presumed motoneurons) were not. NADPH diaphorase-positive cells in the intermediolateral cell column were not immunoreactive with GABA or glycine antibodies. PMID- 8227523 TI - Representation of the body surface in the gracile, cuneate, and spinal trigeminal nuclei of the little red flying fox (Pteropus scapulatus). AB - The body surface representation in the gracile, cuneate, and spinal trigeminal nuclei of the little red flying fox (Pteropus scapulatus) was examined. As in other species, it was found that any single cross-section through all three nuclei contains a representation of most, or all, of the body surface. In the little red flying fox, however, this representation is arranged as a series of dorsolateral to ventromedially oriented bands, within which there are no apparent topographies. These bands are arranged in such a way that the spatial relationships between body regions in the representation do not reflect those at the periphery. PMID- 8227524 TI - Comparative brain ontogeny of the crayfish and clawed lobster: implications of direct and larval development. AB - The freshwater crayfish Cherax destructor and the lobster Homarus americanus have many similarities including life style, body form, and neural organization. However, the ontogenic history is very different in the two species. The development of Cherax is short and direct whereas the development of Homarus comprises three pelagic larval stages and takes more than twice as long from extrusion to benthic stages at constant temperature. In order to determine the progression of maturation of the nervous system in each species and the potential implications of pelagic forms on brain structure, the timing of appearance of 22 general and neural developmental events clearly identifiable in both species was compared. The onset of serotonin antigenicity in the different parts of the brain was chosen as one marker of neural development. During the first month of embryogenesis the timing of morphological, physiological, and neural events is similar in the two species. Morphological development is then accelerated in the crayfish near hatching time and over the two postembryonic stages before the advent of the independent benthic stage. Such heterochronic processes can at least partly account for the different developmental patterns in the two decapods. Among the characters showing similar timing in the two species is the formation of glomeruli (presumptive zones of synaptic contact) in the olfactory lobes of the deutocerebrum, although this event is embryonic in Homarus but postembryonic in Cherax. In contrast, glomerular formation in the accessory lobes is heterochronic: in both species, the glomeruli of the accessory lobes are acquired postembryonically, that is, 3 to 4 months earlier in Cherax than in Homarus. These data suggest that the development of the glomeruli in the olfactory lobes may depend primarily on internal developmental signals, whereas the triggering of glomerular formation in the accessory lobes may depend on external cues. The fact that, in Homarus, only the postlarval stages show mature accessory glomeruli may be a reflection of the functional requirements of benthic life. PMID- 8227525 TI - Distinct subsets of sensory olfactory neurons in mouse: possible role in the formation of the mosaic olfactory projection. AB - The axons of the primary sensory olfactory neurons project from the olfactory neuroepithelium lining the nasal cavity, onto glomeruli covering the surface of the olfactory bulb. Neuroanatomical studies have shown previously that individual olfactory glomeruli are innervated by neurons that are dispersed widely within the nasal cavity. The aim of the present study was to test the hypothesis that phenotypically unique subsets of primary sensory olfactory neurons, scattered throughout the nasal cavity, project to a subset of glomeruli in specific olfactory bulb loci. Immunochemical and histochemical analyses in neonatal mice revealed that the plant lectin, Dolichos biflorus agglutinin, bound to a subset of mature primary sensory olfactory neurons which express the olfactory marker protein. This subset of neurons was principally located in the rostromedial and dorsal portions of the nasal cavity and projected specifically to a subset of glomeruli in the rostromedial and caudodorsal portions of the olfactory bulb. Analysis of Dolichos biflorus-reactive axons revealed that these axons coursed randomly, with no evidence of their selective fasciculation, within the olfactory nerve. It was only at the level of the rostral olfactory bulb that a significant reorganisation of their trajectory was observed. Within the outer fibre layer of the bulb, discrete bundles of lectin-reactive axons began to coalesce selectively into fascicles which preferentially oriented toward the medial side of the olfactory bulb. These data demonstrated that a phenotypically distinct subset of primary sensory olfactory neurons exhibits a topographical projection from the olfactory epithelium to the olfactory bulb, and suggests that these, and other subsets, may form the basis of the mosaic nature of this pathway. Moreover, it appears that the outer nerve fibre layer in the rostral olfactory bulb plays an important instructive role in the guidance and fasciculation of olfactory sensory axons. PMID- 8227526 TI - Regional specificities in the distribution, chemical phenotypes, and coexistence patterns of neuropeptide containing nerve fibres in the human anal canal. AB - Despite the pivotal clinical significance of the human anal canal, little is known about its total and specific innervation. This study assessed the comparative distribution and histotopology of nerve fibres immunoreactive for neural markers and a variety of regulatory active neuropeptides in the human anal canal by light microscopic immunohistochemistry. Depending on the epithelial zone and region of the anal canal, the neural elements were differentially immunoreactive for the pan-neural marker protein gene product 9.5, the catecholamine marker tyrosine hydroxylase, the neuroendocrine marker chromogranin A, and various neuropeptides. Protein gene product 9.5-immunoreactive nerve fibres were ubiquitously abundant in the anal canal. In the anal transitional zone, ectopic epithelial types were supplied by the same pattern of peptidergic nerves as the respective type of epithelium in normotopic location. In the dermis of the squamous zone and in the perianal epidermis, unusual distribution patterns of nerve fibres, referred to as areas of high nerve fibre density, were encountered. Double immunohistochemistry revealed region-specific coexistence patterns of neuropeptidergic nerve fibres, and novel peptide coexistence patterns were detected in anal nerve fibres. Subsets of nerve fibres formed close spatial relationships with chromogranin A-positive neuroendocrine cells, most frequently in the anal transitional zone. Chromogranin-A positive cells were shown to be present in the epithelium of perianal eccrine sweat glands. The differential distribution, peptide phenotypes and coexistence patterns of different nerve fibre populations in the human anal canal may reflect topospecific regulatory functions of neurally released neuropeptides in health and disease. PMID- 8227527 TI - Relationship of descending inferior colliculus projections to olivocochlear neurons. AB - With the objective of defining the relationship of descending inferior colliculus projections to the olivocochlear system in the guinea pig, inferior colliculus neurons were anterogradely labeled with Phaseolus vulgaris-leucoagglutinin and olivocochlear neurons were retrogradely labeled with horseradish peroxidase in the same brain sections. Inferior colliculus neurons were found to project to many nuclei and regions of the hindbrain where olivocochlear neurons reside. The most substantial of these descending projections was to the ipsilateral medioventral periolivary region. Fewer descending projections terminated in the ipsilateral ventral nucleus of the lateral lemniscus, superior paraolivary nucleus, and rostral periolivary region; and even fewer ipsilateral projections terminated in the area surrounding the lateral superior olive, caudal periolivary region, and the lateroventral periolivary region. Descending neurons of the inferior colliculus also project to the contralateral hindbrain first via the lateral lemniscus and then the trapezoid body, to terminate in the contralateral medioventral periolivary region, superior paraolivary nucleus, rostral periolivary region, and the ventral nucleus of the lateral lemniscus. In addition to the projections into these regions that contain olivocochlear neurons, there are varicosities of inferior colliculus neurons that appear to contact the olivocochlear neurons themselves, both ipsilaterally and contralaterally, especially, but not only, in the ipsilateral medioventral periolivary region. We therefore conclude that descending inferior colliculus neurons do provide input to olivocochlear neurons and that the input is not limited to olivocochlear neurons of the ipsilateral medioventral periolivary region. However, given the robust nature of the projection to the ipsilateral medioventral periolivary region and the paucity of contacts observed in that region, we also conclude that the olivocochlear neuron is not the major target of descending inferior colliculus projections. PMID- 8227528 TI - Monoclonal antibody labels olfactory and visual pathways in Drosophila and Apis brains. AB - We employed a monoclonal antibody raised against Drosophila brain homogenate for a comparative immunocytochemical analysis of visual and olfactory pathways in brains of two insect species. On Western blots of Drosophila and Apis nervous tissue, antibody fb45 recognized an antigen with an apparent molecular weight higher than 180 kD. Application of the antibody to sections of Drosophila and Apis brain stained certain interneurons which conspicuously fasciculate in common tracts or neuropilar compartments. Both in Drosophila and in Apis, the antigen was also expressed on the perineural sheath and granular cell compartments in the majority of neuronal cell bodies. The antibody stained monopolar cells in the visual system of both species, and in Apis those fibers of the anterior superior optic tract which link the medulla with the mushroom bodies. In Drosophila, bundles of Kenyon cells of the mushroom bodies were stained. In worker bees and drones, the relay neurons of the median and lateral antennoglomerular tracts were labelled. Since the recognition of the antigen does not require fixation, the antibody can be employed to label selectively living neurons in dissociated cell culture. This opens up the possibility for future functional studies on the role of the antigen in vitro. PMID- 8227529 TI - Nerve growth factor receptor (p75)-immunoreactivity in the normal adult feline trigeminal system and following retrogasserian rhizotomy. AB - The 75 kDa protein nerve growth factor receptor [NGFr(p75)] is a neurotrophin receptor that is able to bind different members of the neurotrophin family of molecules implicated in affecting neuronal survival. Here we describe the light microscopic distribution of NGFr(p75)-immunoreactivity (IR) within the feline trigeminal brainstem sensory nuclear complex and trigeminal ganglion of normal adult subjects and in subjects 10 and 30 days following retrogasserian rhizotomy. Within the trigeminal ganglion of normal subjects, numerous fibers and most of the neuronal cell bodies showed NGFr(p75)-IR that varied in intensity, while cells and fibers with NGFr(p75)-IR were less numerous within the mesencephalic trigeminal nucleus. Within the main sensory and spinal trigeminal nuclei, NGFr(p75)-IR formed a reproducible pattern that varied between the different subnuclei. The NGFr(p75)-IR consisted both of dense pockets and a low level NGFr(p75)-IR that was selective to the trigeminal neuropil. Following rhizotomy, most of the NGFr(p75)-IR was lost from the main sensory and spinal trigeminal nuclei, except in regions where the upper cervical roots and cranial nerves VII, IX, and X project. In contrast, examination of the central root that was still attached to the trigeminal ganglion showed increased NGFr(p75)-IR in fibers and supporting cells, as did the motor root within the peripheral mandibular division. These results indicate that the majority of the NGFr(p75)-IR within the main sensory and spinal trigeminal nuclei originates from primary trigeminal afferents and that retrogasserian rhizotomy leads to an up-regulation of NGFr(p75)-IR in the part of the central root that is contiguous with the ganglion. PMID- 8227530 TI - Spinal termination patterns of canine identified A-delta and C spermatic polymodal receptors traced by intracellular labeling with Phaseolus vulgaris leucoagglutinin. AB - The spinal projection patterns of spermatic polymodal receptors were studied by intracellular labeling of functionally identified canine dorsal root ganglion (DRG) neurons with Phaseolus vulgaris-leucoagglutinin (PHA-L). The processes of 2 C-fiber and 1 A-delta-fiber spermatic polymodal receptor neurons were labeled well enough to trace their central terminations. The labeled C-fiber DRG neurons were of medium size (mean diameter 36.8 and 40.7 microns). On entering the spinal cord, axons of the C-polyclonal receptors divided into rostral and caudal main branches that extended over 3 spinal segments (20 and 25 mm, respectively), and issued a total of 16 and 15 collaterals, respectively. The majority of collaterals ran in or along the lateral surface, but both neurons had 1 or 2 collaterals or terminal branches running through the middle, or along the medial surface of the dorsal horn. Terminal swellings and en passant enlargements were observed mainly in laminae I, V, and VII. Some C-fiber terminations appeared in lamina II and the adjacent lateral column. The A-delta polymodal receptor had a termination pattern similar to that of the C-fiber units with the exception of a shorter distance over which its 13 identified collaterals were issued (10 mm), and continuation of the rostral main branch into Lissauer's tract (traced for 3.6 mm) after all branches appeared. Two terminal branches were found running just above the central canal in another A-delta neuron in which termination could be only partially traced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227531 TI - Retrograde labeling of neurons in the spinal cord that project directly to the amygdala or the orbital cortex in the rat. AB - The amygdala and orbital cortex are thought to play an important role in the regulation of autonomic functions, hormonal secretion, and behavioral expression in response to sensory stimulation. The responsiveness of neurons in these regions to stimulation of cutaneous and visceral organs indicates that sensory information reaches the amygdala and orbital cortex. In the past, a large number of studies have thoroughly documented multiple neural pathways by which sensory information can reach these regions via relay nuclei in the brainstem and diencephalon. Recent studies reported that the amygdala and orbital cortex also receive direct input from the spinal cord. The aim of this study was to determine the magnitude and the origin of these projections in the rat. Injections of the retrograde tracer Fluoro-Gold (FG), restricted to the amygdala, labeled several hundred neurons bilaterally (60% contralateral) throughout the length of the spinal cord. More than 60% of labeled neurons were found in the lateral reticulated area of the deep dorsal horn and the gray matter surrounding the central canal. Many neurons were also found in the lateral spinal nucleus. Labeled neurons were concentrated in upper lumbar and upper cervical segments. Injections of Fluoro-Gold that were centered in the orbital cortex labeled only a small number of neurons (73% contralateral) within the spinal cord. Most labeled neurons were found in the lateral reticulated area. Neurons located in the intermediate zone and the gray matter surrounding the central canal were found mainly in upper lumbar and upper cervical segments. These findings, together with the anterograde tracing observations, provide evidence for direct projections of spinal cord neurons to the amygdala and orbital cortex. Their laminar distribution in the spinal cord and the involvement of the amygdala and orbital cortex in limbic functions suggest that these pathways may play a role in neuronal circuits that enable somatosensory information, including pain, to affect autonomic, endocrine and behavioral functions. PMID- 8227532 TI - Connections of the olfactory bulb in the gymnotiform fish, Apteronotus leptorhynchus. AB - This work examines the connectivity of the olfactory bulb in the gynmotiform fish Apteronotus leptorhynchus. Wheat germ agglutinin conjugated horseradish peroxidase was iontophoresed in different areas and depths of the bulb in order to define its efferent and afferent connections. The olfactory bulb projects bilaterally via the medial (medial and centromedial fascicles) and lateral olfactory (lateral and centrolateral fascicles) tracts. The nervus terminalis courses through the ventromedial aspect of the bulb to terminate in parts of the medial subpallium and hypothalamus. Its telencephalic component could be identified by a nonpreadsorbable substance P-like immunoreactivity. Fibers within the medial olfactory tract form four telencephalic terminal fields: peduncular, medial, intermediate and posterior fields. The diencephalic terminal fields in the habenula, preoptic, and hypothalamic areas appear to correspond to some of the nervus terminalis fibers (von Bartheld and Meyer [1986] Cell Tissue Res. 245:143-158, Krishna et al. [1992] Gen. Comp. Endocrinol. 85:111-117), and to axons of telencephalic bulbopetal cells of area dorsalis posterior. The terminal fields of the medial olfactory tract and nervus terminalis partially overlap in the ventral telencephalic areas partes ventralis, supracommissuralis, and rostral preoptic region. The lateral olfactory tract forms a lateral terminal field and contributes to the intermediate and posterior terminal fields. Olfactory fibers cross in the interbulbar, anterior, and habenular commissures and tuberal decussation. Consistent differences were noted between the medial and lateral olfactory bulb, with respect to their cytoarchitectonics, immunohistochemistry, and connections. In addition to the olfactory nerve, bulbar afferents are predominantly ipsilateral, with minor inputs originating from the contralateral bulb and telencephalic area dorsalis posterior, nucleus raphe centralis, and locus ceruleus. PMID- 8227533 TI - Fate of abdominal ventral unpaired median cells during metamorphosis of the hawkmoth, Manduca sexta. AB - Each of the unfused abdominal ganglia in the larval, pupal, and adult stages of the hawkmoth, Manduca sexta, has two large ventral median neurons with axons that bifurcate to innervate targets on both sides of the abdomen. Although the dendritic structures of the two neurons are similar, their axons branch to innervate distinct sets of target muscles. During metamorphosis both neurons undergo dendritic regression, followed by growth of new arborizations during adult development. The neurons must innervate different targets in the larva and adult, since many larval muscles degenerate and are replaced during metamorphosis. Both neurons were reactive with an antibody to the neuromodulatory compound, octopamine, in the larval and adult stages. Pairwise intracellular recordings in isolated nerve cords revealed spontaneous excitatory synaptic potentials that occurred in the ventral median neurons of each ganglion in an anterior-to-posterior sequence. The synaptic potentials were eliminated when the interganglionic connective was interrupted posterior to the subesophageal ganglion. The ventral median neurons were also excited by tactile stimulation of the body surface in larvae, pupae and adults. PMID- 8227534 TI - Immunocytochemical localization of glycine receptors in the mammalian retina. AB - The distribution of glycinergic synapses in the mammalian retina was studied with monoclonal antibodies against glycine receptors and a glycine receptor-related protein (gephyrin). Monoclonal antibody 2b is specific for the alpha 1 subunit of the glycine receptor; monoclonal antibody 4a is specific for all known alpha subunits and the beta subunit, and monoclonal antibody 7a is specific for gephyrin. The three antibodies were applied to the retina of cat, macaque monkey, rat, and rabbit. The general staining pattern is comparable in all these species and it is similar but distinct with all of the three antibodies. Labeling is characterized by a punctate appearance indicating that it occurs at synapses. In the inner plexiform layer, labeling is concentrated in two bands. One band is located close to the inner nuclear layer; the other band is located in the middle of the inner plexiform layer. In the outer plexiform layer, sparse punctate labeling is seen. The distribution of gephyrin was also studied at the ultrastructural level in cat and monkey retina. Gephyrin is present on the postsynaptic membrane of amacrine cells and ganglion cells. The presynaptic profile to gephyrin immunoreactivity is always of an amacrine cell. The AII amacrine cell, the crucial glycinergic interneuron of the rod pathway, is presynaptic to gephyrin immunoreactivity in the OFF-sublamina and is itself gephyrin-positive at an input synapse from another (possibly GABAergic) amacrine cell in the ON-sublamina. PMID- 8227535 TI - Topography of cholinergic afferents from the nucleus basalis of Meynert to representational areas of sensorimotor cortices in the rat. AB - We investigated (1) the topography of projection neurons in the nucleus basalis of Meynert (NBM) with efferents to restricted regions of the primary somatosensory (SI), the second somatosensory (SII), and the primary motor (MI) cortices in the rat; (2) the percentage of these NBM projection neurons that were cholinergic; and (3) the collateralization, if any, of single NBM neurons to different subdivisions within SI, to homotopic areas of SI and SII, and to homotopic areas of SI and MI. Retrograde single- and double-labeling techniques were used to study NBM projections to electrophysiologically identified subdivisions of SI and to homotopic representational areas of SI and SII, and of SI and MI. Choline acetyltransferase immunocytochemistry was done to identify cholinergic NBM neurons. Of the retrogradely labeled NBM neurons that projected to selective subdivisions of SI, SII, and MI, 89%, 87%, and 88%, respectively, were cholinergic. We found a rostral-to-caudal progression of retrogradely labeled NBM neurons following a medial-to-lateral sequence of injections into subdivisions of SI. Overlapping groups of single-labeled NBM neurons were observed after injections of different tracers into adjacent subdivisions within SI or homotopic areas of SI and SII, and of SI and MI. We conclude that NBM innervation to SI, SII, and MI is mostly cholinergic in the rat, that each cortical area receives cholinergic afferents from neurons widely distributed within the NBM, and that each NBM neuron projects to a restricted cortical area without significant collateralization to adjacent subdivisions within SI or to homotopic areas of SI and SII, or SI and MI. PMID- 8227536 TI - Long-term consequences of impaired regeneration on facial motoneurons in the C57BL/Ola mouse. AB - Peripheral nerves of the C57BL/Ola mouse mutant undergo markedly slowed Wallerian degeneration following injury. This is associated with impaired regeneration of both sensory and motor axons. Following a crush lesion of the facial nerve, there was no cell loss in facial nuclei of normal (C57BL/6J) adult mice, but 40% cell loss occurred in Ola mice and the survivors increased in size during the period when functional reinnervation was established. These results are interpreted as a result, first, of prolonged deprivation of target-derived trophic factor in the slowly regenerating Ola motoneurons and second, increased peripheral field size of the survivors. Within the regenerated facial nerve, there was marked heterogeneity of myelinated fibre size in Ola mice. Some Ola axons, both proximal and distal to the lesion site, had areas over twice as great as the largest 6J axons when measured 1 year following injury. A population of small diameter fibres, not observed in 6J nerves, persisted distal to the crush site in Ola nerves, and this was associated with an increase in the total number of myelinated axons in the distal nerve: on average, each parent Ola axon retained three persistent daughter axons. The delayed Wallerian degeneration in Ola mice not only impairs immediate axon regrowth, but also results in a breakdown of the normal mechanisms which regulate axon number and size in regenerating nerve. PMID- 8227537 TI - Merkel-like basal cells in Necturus taste buds contain serotonin. AB - Several types of cells have been identified in vertebrate taste buds, including dark cells, light cells, intermediate cells, type III cells, and basal cells. The physiological roles of these cell types are not well understood, especially those of basal cells. In this paper we show that there are two types of basal cells in taste buds from Necturus maculosus. One type of basal cell is an undifferentiated cell, presumably a stem cell. By combining light microscopic immunocytochemistry with electron microscopy, we show that the other type of basal cell is positive for serotonin-like immunoreactivity and that these cells have ultrastructural features similar to those found in cutaneous Merkel cells. Based on these findings, and the fact that the Merkel-like taste cells have been shown to make synaptic contacts with adjacent taste cells and with innervating nerve fibers, we conclude that these Merkel-like basal taste cells are serotonergic interneurons. PMID- 8227538 TI - Facial cutaneous reconstructive surgery: general aesthetic principles. AB - The performance of cutaneous reconstructive surgery requires understanding and application of many important principles. This article reviews the critical factors to consider in the management of surgical wounds by second-intention healing, primary closure, skin grafting, and repair with local flaps. For certain defects, reconstruction with local flaps offers several advantages over other alternatives. Key concepts useful in flap choice and implementation are discussed, and surgical techniques that maximize the aesthetic outcome of reconstructive surgery are reviewed. PMID- 8227539 TI - Cutaneous manifestations of Epstein-Barr virus-associated T-cell lymphoma. AB - BACKGROUND: In addition to human T-lymphotropic virus (HTLV-I), the Epstein-Barr virus (EBV) has recently been demonstrated to be associated with cutaneous T-cell lymphoma (CTCL). OBJECTIVE: Our purpose was to investigate characteristic clinicopathologic features of the cutaneous lesions of EBV-associated T-cell malignancies. METHODS: Clinical records, laboratory data, and histopathologic sections were reviewed. Freshly frozen tumor tissues were immunophenotyped. Southern blot and in situ hybridization studies were performed to detect the EBV genomes. RESULTS: Ten of 35 CTCL biopsy specimens collected between 1985 and 1992 were found to be EBV-associated. Clonotypic proliferation of EBV genomes was demonstrated in each case, and the atypical T lymphoid cells contained EBV genomes. The cutaneous eruptions of these patients included multiple violaceous papules or nodules, chronic ulcers, and tumors on the trunk or extremities. Three distinct clinicopathologic subgroups could be recognized. The most consistent was the angiocentric T-cell lymphoma or lymphomatoid granulomatosis (type III CTCL) (four cases), presenting with chronic ulcers or violaceous papules. The second group was the T large-cell lymphoma (type II CTCL), Ki-1 antigen (CD30) (positive or negative) (four cases). Three patients with Ki-1- lymphoma had fulminant disease, whereas the remaining Ki-1+ case had a benign course. The third group was the secondary type CTCL (type V CTCL) (two cases), representing systemic EBV associated T-cell lymphoma. The prognosis was grave. The common features of these EBV-associated CTCLs are resistance to conventional chemotherapy, poor prognosis, and the terminal manifestation of a hemophagocytic syndrome. No EBV genome could be detected in 12 cases of classic CTCL/mycosis fungoides (type I CTCL), or in three cases of HTLV-I-associated adult T-cell lymphoma (type IV CTCL). CONCLUSION: Three distinct clinicopathologic subtypes of EBV-associated CTCL were recognized, including one additional type of virus-associated CTCL. PMID- 8227540 TI - Partial unilateral lentiginosis. AB - BACKGROUND: We review our experience with nine patients with partial unilateral lentiginosis (PUL), a rare pigmentary disorder. OBJECTIVE: Our purpose was to define the characteristics of PUL and to discuss the differential diagnosis. METHODS: The records of nine patients with PUL were reviewed. A literature review on diagnosis, association with other disorders, and differential diagnosis is presented. RESULTS: PUL is a rare benign disorder that has no known inheritance pattern and has no commonly associated abnormalities. CONCLUSION: Careful history and physical examination may enable the distinction between PUL, nevus spilus, and other more serious genetic disorders associated with lentiginosis. PMID- 8227541 TI - Ki-1-positive anaplastic large-cell lymphoma can mimic benign dermatoses. AB - BACKGROUND: Regressing atypical histiocytosis is a recently described disease characterized by recurrent nodules or ulcers. The cutaneous lesions appear abruptly and then regress only to return in a manner reminiscent of lymphomatoid papulosis. Immunophenotypic analysis has revealed that most cases are a form of anaplastic large-cell Ki-1-positive (CD30+) lymphoma. OBJECTIVE: We describe two patients with Ki-1-positive anaplastic large-cell lymphoma that had clinical and pathologic features of regressing atypical histiocytosis and mimicked benign dermatoses (pyoderma gangrenosum and morphea), causing a delay in confirming the true diagnosis. A third case that was readily recognized as a lymphoma is also presented. METHODS: The clinical and histopathologic findings were recorded. In addition, T-cell receptor gene rearrangement and immunophenotyping were determined in the index case. RESULTS: The index patient and second patient were diagnosed as having Ki-1-positive anaplastic large-cell lymphoma by immunophenotyping and underwent cyclophosphamide, doxorubicin, prednisone, and vincristine (CHOP) chemotherapy with complete remission. The patient detected by chart review died of her disease without receiving antineoplastic therapy; disseminated lymphoma was diagnosed at autopsy. Studies on paraffin-embedded tissue were consistent with Ki-1-positive anaplastic large-cell lymphoma. CONCLUSION: Regressing atypical histiocytosis may clinically resemble some benign dermatoses. Recent evaluation of these cases has shown that many represent a form of Ki-1-positive anaplastic large-cell lymphoma. Multiple skin biopsy specimens with immunophenotyping and gene rearrangement studies are required to arrive at the diagnosis. PMID- 8227542 TI - Acquired ichthyosis in concomitant HIV-1 and HTLV-II infection: a new association with intravenous drug abuse. AB - BACKGROUND: Papulosquamous eruptions are common in HIV-1-infected patients. Acquired ichthyosis may occur after profound T-cell depletion. Intravenous drug users infected with HIV-1 can be coinfected with human lymphotropic virus II (HTLV-II). OBJECTIVE: We examined the relation between acquired ichthyosis and concomitant infection with HIV-1 and HTLV-II in intravenous drug users. METHODS: We examined 184 male and female HIV-1-positive intravenous drug users for acquired ichthyosis. Enzyme-linked immunosorbent assay was used to screen these patients for antibody to HTLV-I/II. Western blot, viral isolation, and the polymerase chain reaction were used to confirm that serologic responses were from HTLV-II and not HTLV-I. RESULTS: Acquired ichthyosis occurred in 6.3% of white, 16.4% of Hispanic, and 21.7% of black patients. It occurred only after profound helper T-cell depletion, in association with increasing age, and with concomitant infection with HTLV-II (22.2% vs. 6.8% in HIV-1 singly infected patients [p < 0.038]). CONCLUSION: Acquired ichthyosis may be a marker of concomitant infection with HIV-1 and HTLV-II in intravenous drug users and occurs after profound helper T-cell depletion. PMID- 8227543 TI - Hemodynamics of leg ulceration assessed by laser Doppler flowmetry. AB - BACKGROUND: Cutaneous blood flow and the venoarteriolar reflex can be assessed in the leg by laser Doppler flowmetry. OBJECTIVE: Our purpose was to compare these measurements in ulcerated and nonulcerated skin in venous and arterial disease and to determine whether either changes with healing. METHODS: Studies were performed in 20 patients with venous disease, 16 with arterial disease, and 12 control subjects and repeated sequentially in 15 patients with small venous ulcers until they healed. RESULTS: Blood flow in nonulcerated skin was greatest in venous disease (but not when severe ulceration was excluded). The reflex was exaggerated in nonulcerated skin in venous disease but attenuated in the base of both venous and arterial ulcers. The reflex reappeared in the base of the venous ulcers as they healed. CONCLUSION: Measurement of the venoarteriolar reflex in nonulcerated skin distinguished between venous and arterial disease and its reappearance in the base of venous ulcers was associated with healing. PMID- 8227544 TI - Merkel cell carcinoma: in vitro and in vivo characteristics of a new cell line. AB - BACKGROUND: Few studies exist that describe Merkel cell carcinoma (MCC) growth characteristics in vitro, in vivo, or both. OBJECTIVE: Our purpose was to evaluate the pathologic features of MCC implanted into athymic mice and to determine cytogenetic abnormalities in the established cell line. METHODS: Tumor tissues from a patient with MCC were grown in culture. Cultured cells were karyotyped and inoculated subcutaneously into athymic mice. Nude mouse tumors were re-implanted into other athymic mice. Tissues from the primary skin tumor and the nude mouse tumor were processed for light and electron microscopy and immunocytochemistry. RESULTS: The cell line showed a doubling time of 64.8 hours. Xenografts of 4 x 10(6) cells produced tumors in athymic mice with a doubling time of 16.1 days. The nude mouse tumors showed pathologic features similar to those of the primary skin tumor. Cytogenetic studies showed a t(1;17) (p36;q21) translocation in 100% of the cells. CONCLUSION: MCC implanted into athymic mice retained the pathologic features of the primary skin tumor and behaved aggressively. The t(1;17) (p36;q21) translocation may be a marker of an aggressive phenotype. PMID- 8227545 TI - alpha-Amylase, a flour additive: an important cause of protein contact dermatitis in bakers. AB - BACKGROUND: alpha-Amylase, an enzyme commonly used in flour additives, has been reported to be an important cause of rhinitis and asthma in bakers. OBJECTIVE: Our purpose was to determine whether this enzyme could also cause dermatitis. We tested it routinely in bakers with hand eczema. METHODS: Patch tests were administered with the International Contact Dermatitis Research Group standard series and a bakery series and scratch-chamber or prick tests were performed with the bakers' own material and with alpha-amylase powder. RESULTS: Of 32 bakers tested, seven had an immediate wheal-and-flare reaction and two also had a delayed eczematous reaction. High dilutions of the alpha-amylase powder still gave strong reactions. CONCLUSION: alpha-Amylase is an important cause of skin reactions in bakers and should be tested routinely if a contact allergy is suspected. PMID- 8227546 TI - Comparison of narrow-band (311 nm) UVB and broad-band UVA after oral or bath water 8-methoxypsoralen in the treatment of psoriasis. AB - BACKGROUND: There is a disparity between the absorption spectrum of 8 methoxypsoralen and the action spectrum for psoralen-sensitized erythema. In an action spectrum corrected for unsensitized reaction 313 and 365 nm have similar efficacies. OBJECTIVE: We evaluated the relative erythemogenic and antipsoriatic efficacy of narrow-band (311 nm) UVB with and without prior psoralen exposure. We also compared the effects of narrow-band UVB and broad-band UVA after oral and bath-water psoralen exposure. METHODS: Patients with psoriasis underwent half side comparison studies. In one group the therapeutic efficacy of 311 nm UVB with and without oral psoralen was assessed. The second group received UVA and 311 nm UVB after oral psoralen. The third group was exposed to both radiation sources after bath-water exposure. RESULTS: The erythemogenic, pigmentogenic, and therapeutic efficacy of 311 nm was increased by oral psoralen. With systemic 8 methoxypsoralen, UVA was comparable to 311 nm UVB. After bath-water exposure, 311 nm was clearly superior to broad-band UVA. CONCLUSION: The efficacy of narrow band 311 nm UVB can be enhanced by psoralen. Narrow-band 311 nm UVB is also effective after psoralen bath-water delivery. PMID- 8227547 TI - Expression of p53 protein in benign and malignant epidermal pathologic conditions. AB - BACKGROUND: p53 is an oncogene and a tumor-suppressor gene whose gene product regulates the cell growth cycle. Mutations in p53 are the most common genetic alterations in human cancer. OBJECTIVE: Our purpose was to define p53 expression and subcellular localization in normal and pathologic epidermis. METHODS: We examined sections of normal skin, psoriasis, squamous cell carcinoma (SCC), basal cell carcinoma (BCC), keratoacanthoma, and cultured epithelial cell lines with five antibodies to p53. RESULTS: Monoclonal antibody MAb421 and MAb1801 stained the cytoplasm of normal basal keratinocytes, suggesting that subcellular localization or sequestration of wild-type p53 regulates its activity. Polyclonal antibody reacted throughout normal epidermis, suggesting heterogeneity of species or conformational forms of p53 protein. Cytoplasmic reactivity to MAb421 was similar in normal epidermis, psoriasis, and cultured keratinocytes but was, diminished in SCC, BCC, and keratoacanthoma. CM-1 reactivity persisted in these tumors. Putative p53 mutations detected by MAb240 reactivity were present in 44% of SCC specimens. CONCLUSION: Epidermis expresses p53 and the gene may regulate epidermal keratinocyte growth and carcinogenesis. PMID- 8227548 TI - Oral manifestations in HIV-infected patients: diagnosis and management. AB - Oral lesions have been observed since the beginning of the AIDS epidemic. The number of HIV-infected patients is still increasing, especially in the heterosexual population. Oral diseases in HIV-infected patients are often more difficult to diagnose because the clinical presentations may differ from the same diseases in HIV-negative patients. HIV-associated oral lesions have diagnostic, prognostic, and therapeutic impact. Approximately 10% of the HIV-infected population will have oral manifestations as a first sign of their disease. In HIV infected men oral hairy leukoplakia and oral candidiasis are useful markers for disease progression. This article summarizes the oral manifestations and the management of oral health in persons with HIV infection. PMID- 8227549 TI - 1992 AAD Award for Excellence in Education: The Integrated Basic and Clinical Science Conference Series at the University of Texas Southwestern Medical Center. AB - This article was written to share information concerning the establishment and implementation of a successful, integrated basic and clinical science conference series for resident trainees in dermatology. We discuss the underlying philosophy and describe the concepts employed in, and the dynamics that have resulted from, this educational experiment. Finally, we include the schedule and references for the 1993-1994 academic year. PMID- 8227550 TI - The diminishing role of the dermatologist in the office-based care of cutaneous diseases. AB - The number of physicians who provide ambulatory care for patients with skin diseases is rapidly increasing. Current and proposed changes in health care financing may limit direct access to dermatologists. We provide recent data on volume of visits for cutaneous diagnoses, including the types of medical practitioners who provided these ambulatory services, and compare these data with earlier surveys that span 15 years. In 1989, dermatologists saw 41% of the 51.6 million persons with cutaneous diagnoses. The number of visits per dermatologist declined significantly from 1974 to 1989 (p < 0.05). Visits to dermatologists were significantly more likely to be reimbursed by Blue Cross/Blue Shield or self pay than visits to other physicians for skin problems. From 1974 to 1989, the demand for dermatologic services from dermatologists has not increased as rapidly as the supply of dermatologists. Our data also suggest that restrictions on direct access to dermatologists or changes in reimbursement may substantially decrease the demand for services provided by dermatologists. PMID- 8227551 TI - Skin involvement in giant lymph node hyperplasia (Castleman's disease). PMID- 8227552 TI - Basal cell carcinoma and keratoacanthoma in Hawaiians: an incidence report. PMID- 8227553 TI - Female pattern baldness in men. PMID- 8227554 TI - Brachioradial pruritus: treatment with topical capsaicin. PMID- 8227555 TI - Successful treatment of albopapuloid epidermolysis bullosa (Pasini's variant) with pulse topical corticosteroid therapy. PMID- 8227556 TI - Tanning salon porokeratosis. PMID- 8227557 TI - Incomplete Reiter's syndrome induced by systemic interferon alpha treatment. PMID- 8227558 TI - Treatment of angiofibroma with the pulsed tunable dye laser. PMID- 8227559 TI - Seborrheic keratoses with occult underlying basal cell carcinoma. PMID- 8227560 TI - Chronic meningococcemia in acquired immunodeficiency infection. PMID- 8227561 TI - Zosteriform transient acantholytic dermatosis. PMID- 8227562 TI - Unilateral laterothoracic exanthem in children. PMID- 8227563 TI - Cutaneous markers of internal malignancy. PMID- 8227565 TI - Self-assessment examination of the American Academy of Dermatology. Skin papules and nodules. PMID- 8227564 TI - Self-assessment examination of the American Academy of Dermatology. Leg ulcer. PMID- 8227566 TI - Interobserver variability in CT and MR staging of lung cancer. AB - OBJECTIVE: Our goal was to assess the interobserver variability in staging non small cell lung cancer using CT and MRI. MATERIALS AND METHODS: As part of the Radiologic Diagnostic Oncology Group (RDOG) study of lung cancer staging, the CT and MR examinations of 40 patients suspected of having non-small cell bronchogenic carcinoma were blindly interpreted by four expert observers. The primary tumor and lymph node stages in the 40 study subjects were similar to the final proportions reported in the RDOG study. Assessed abnormalities included the presence of a lung nodule, chest wall invasion, mediastinal invasion, bronchial involvement, lymph node metastasis in specific node stations, and T and N classifications. Percent agreement and kappa-values were calculated for each of these determinations. RESULTS: Depending on the finding assessed and the method of analysis, average agreement rates ranged from 58 to 90% for CT and from 61 to 96% for MRI. Average kappa-values were largely between 0.40 and 0.60 when dichotomous analysis was used; weighted kappa-values were similar. With a single exception, no significant differences were found for kappa-values calculated for CT and MRI. CONCLUSION: Although interobserver agreement rates are good for determining T and N classification in patients with lung cancer, variability in image interpretation is frequent, even among experienced observers. PMID- 8227567 TI - Comparison of fixed and adjustable window width and level settings in the CT evaluation of diffuse lung disease. AB - OBJECTIVE: The purpose of this investigation was to assess the utility of allowing free adjustment of window width and level in comparison with the use of a fixed lung window in the CT evaluation of diffuse lung disease. MATERIALS AND METHODS: Six radiologists each judged 36 cases (28 diffuse lung disease and 8 normal) using a standardized form. In half of the sessions, images were viewed in a fixed lung window (level = -500 HU; width = 2,000 HU). In the other sessions, the observer was able to adjust the window and level freely while viewing the images. Each case was seen twice in separate sessions: once in a fixed lung window and once with window width and level adjusted by the reader. A variety of diagnostic features were evaluated using a 5 point scale. These included visibility of fine lung structures, abnormalities of the lung parenchyma, and overall evaluation of the lung. RESULTS: The visibility of lung structures was not improved with adjustable window settings. Receiver operating characteristic (ROC) analysis showed fixed windows to be superior to adjustable windows for overall evaluation of the cases [fixed A(z) = 0.90, adjustable A(z) = 0.84, p < 0.05, jackknife method]. Time to rate each case was increased by 15% with window width and level adjustment. CONCLUSION: Free adjustment of window width and level produced no improvement in reader performance over that achieved with fixed window width and level. PMID- 8227568 TI - Bronchiolitis obliterans on high-resolution CT: a pattern of mosaic oligemia. AB - OBJECTIVE: Bronchiolitis obliterans, when not associated with organizing intraalveolar pneumonia or extensive peribronchiolar fibrosis, is often difficult to distinguish clinically and radiographically from other forms of chronic obstructive pulmonary disease. The aim of this study was to demonstrate a pattern on high-resolution CT (HRCT) that could suggest this diagnosis. MATERIALS AND METHODS: Two patients with a clinical diagnosis of bronchiolitis obliterans, moderate-to-severe obstruction on pulmonary function tests, and normal chest radiography were studied with HRCT. RESULTS: High-resolution CT of the patients with chronic bronchiolitis demonstrated a mosaic pattern of low attenuation probably corresponding to secondary lobules. CONCLUSION: We conclude that the low attenuation areas represent either air trapping or hypoxic vasoconstriction in secondary pulmonary lobules from obstruction of small airways. This pattern of mosaic oligemia was, until recently, only recognized with occlusive vascular disease and may suggest obstructive disease in the small airways. PMID- 8227569 TI - Systemic arterial supply to normal basal segments of left lower lobe: CT findings. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of CT in making a diagnosis of systemic arterial supply to normal basal segments of the left lower lobe. MATERIALS AND METHODS: We retrospectively reviewed chest radiography (three cases), CT (three cases), and angiography (two cases). RESULTS: An anomalous systemic artery arising from the descending aorta was shown as a retrocardiac density on posteroanterior radiography and as a nodular density behind the heart on lateral radiography. Contrast enhanced CT revealed the vascular nature of the density adjacent to the descending aorta. Dynamic CT in one patient demonstrated that the nodular density was composed of an anomalous systemic artery and a prominent inferior pulmonary vein. High-resolution CT showed the interlobar artery distal to the origin of the superior segmental artery to be absent. The bronchial system of the left basal segments was normal as was the pulmonary parenchyma on CT. CONCLUSION: Characteristic CT findings consist of absence of the interlobar artery distal to the origin of the superior segmental artery and origination of an anomalous artery from the descending aorta that gives off branches to normal left basal segments. With a constellation of these CT findings, angiography would be obviated for definite diagnosis of this anomaly. PMID- 8227570 TI - MRI in hypertrophic cardiomyopathy: a morphofunctional study. AB - OBJECTIVE: We compared MRI with two-dimensional echocardiography (2dE) and Doppler echocardiography to determine the diagnostic role of MRI in hypertrophic cardiomyopathy (HCM). MATERIALS AND METHODS: Twenty-three patients with 2dE diagnosis of HCM were examined with MRI; 12 of 23 patients were also studied by color (cDE) and continuous wave (cwDE) Doppler echocardiography. Morphologic information and diastolic heart wall thickness were obtained by SE sequences; functional study was performed by gradient echo sequences (cine MR). RESULTS: The correlation between MR, SE sequences and 2dE was better for septal (r = 0.930, p < 0.01) than for posterolateral (r = 0.739, p < 0.01) wall thickness. The assessment of the distribution of the hypertrophy was changed by MR in five cases. Cine MR functional study showed a systolic subaortic signal void (dynamic obstruction) in 12 of 22 patients and a systolic left atrial signal void (mitral regurgitation) in 17 of 22. Systolic wall thickening was studied by cine MR and 2dE in 11 patients: A good correlation was found for septum (0.01 < p < 0.05) and a poor one for posterolateral wall (p > 0.05). The cine MR and cDE turbulence duration in the left ventricle and atrium showed excellent correlation (p < 0.01). Good agreement was found between the duration of subaortic turbulence (cine MR or cDE) and the pressure gradient (cwDE) (p < 0.01 and 0.01 < p < 0.05, respectively) and between cine MR and cDE semiquantitative estimate of the mitral regurgitation (p < 0.01). In all patients with subaortic MR signal void studied with cwDE, a pressure gradient was present. CONCLUSION: Magnetic resonance imaging can play an important role in the diagnosis of HCM after 2dE-DE. PMID- 8227571 TI - Postoperative MR follow-up of type A aortic dissection. AB - OBJECTIVE: We set out to study the efficacy of MRI in the demonstration of residual alterations and postsurgical complications of type A aortic dissection. MATERIALS AND METHODS: From October 1988 to December 1990, 28 patients, 18 patients with type I and 10 with type II aortic dissection, underwent MR examinations for postsurgical evaluation. Features evaluated included caliber of the aorta (a) above and (b) below the prosthesis; (c) redissection; (d) persistent intimal flap; (e) presence of abdominal vessels arising from the false lumen; and (f) status of thoracic supraaortic vessels. RESULTS: Five of 28 patients were considered "normal," due to negative results of the assessed parameters. Dilatation of the aorta was found distal to the graft in seven patients and proximal to the graft in two. Redissection was observed in 2 of 28 patients. Residual intimal flap was seen in 15 of 28 patients; the state of false lumen was well identified in 5 patients using only SE images and in 7 of the remaining 10 using phase imaging. In 25 of 28 patients, supraaortic vessels were well evaluated with involvement seen in 2 cases. In the three other patients, adequate identification was not possible. CONCLUSION: We believe MRI is the technique of choice for monitoring the aorta after surgery in aortic dissection to identify alterations and complications and institute suitable therapy. PMID- 8227572 TI - MRI in pre- and postoperative assessment of tracheal stenosis due to pulmonary artery sling. AB - OBJECTIVE: To evaluate the potential of MRI in the assessment of tracheal stenosis due to tracheal or vascular malformations, 45 children with severe respiratory distress were examined prospectively during a period of 1 year. Five of these children had tracheal stenosis due to a sling left pulmonary artery (SLPA). MATERIALS AND METHODS: Magnetic resonance examinations of the anesthetized children were performed with a 1.5 T Siemens MR imager using electrocardiographically gated T1-weighted SE sequences in transverse and sagittal slice orientations. Slice thickness was 3 mm and each sequence was repeated after shifting the slice position by 1 mm. Monitoring during the examinations included ECG, oscillatory blood pressure, respiratory rate, and oxygen concentration. Magnetic resonance findings were compared with esophagography, selective pulmonary angiography, bronchoscopy, Doppler sonography, and surgery. All examinations were repeated after surgical therapy to assess the improvement in tracheal stenoses and the patency of the ligated and reimplanted left pulmonary arteries. RESULTS: Magnetic resonance imaging clearly revealed the course of the SLPA and its topographic relationship to the trachea as well as the coexistence of cardiovascular and tracheobronchial or esophageal malformations. The degree and length of tracheal stenoses, which were measured in the pre- and postoperative axial slices and graphically displayed, as well as the angles of the right and left main stem bronchi, could be accurately determined. CONCLUSION: Magnetic resonance imaging in combination with bronchoscopy yielded the necessary and sufficient information for diagnosis and aided the surgeon in planning operative strategy and in postoperative follow-up. PMID- 8227573 TI - 3D image reconstruction of right subclavian artery aneurysms. AB - OBJECTIVE: Three-dimensional (3D) image reconstruction methods allied to spiral CT (SCT) or MR angiography (MRA) are used in clarifying the anatomy of complex aortic anatomy. MATERIALS AND METHODS: Two patients with superior mediastinal masses suggestive of aneurysms of the anomalous right subclavian artery were examined. Both patients were examined using breath-hold SCT and one by breath hold MRA. Three-dimensional images were reconstructed using a surface rendering technique for the SCT examinations and by a maximum intensity projection technique for MRA. RESULTS: Two cases of aneurysmal anomalous right subclavian artery were diagnosed by SCT with 3D image reconstruction. One case was also examined by MRA with 3D image reconstruction which confirmed the findings of SCT. The use of the 3D techniques clarified the complicated anatomy and avoided the need for angiography. CONCLUSION: Two cases are illustrated of 3D imaging of complicated aortic branch anatomy due to aneurysms of anomalous right subclavian artery. Image quality was good and no further imaging by more invasive techniques was required. PMID- 8227574 TI - Contrast-enhanced MRI of the breast after limited surgery and radiation therapy. AB - OBJECTIVE: Posttherapeutic changes in the breast after tumorectomy (TE) and radiation therapy (RT) may mimic or obscure recurrent or new malignancies and thus interfere with conventional diagnostic studies. We investigated the enhancement of tissue during variable time intervals after therapy with contrast enhanced MRI in 62 patients. MATERIALS AND METHODS: We report the results of 77 studies in 62 patients undergoing TE and RT. We include only those studies with at least 24 months of clinical and mammographic follow-up (n = 60) or histopathologic results (n = 17). RESULTS: Up to 9 months after therapy, differentiation between posttherapeutic changes and recurrence was frequently impossible because of the strong enhancement. Ten to 18 months after therapy, this posttherapeutic enhancement subsided slowly with some interindividual variations. After 18 months posttherapy, no significant enhancement was encountered in 30 of 32 cases. Diffuse or focal enhancement was present in all recurrent tumors and all recurrences were correctly diagnosed. Furthermore, 4 of 11 recurrences and 10 of 18 single recurrent foci were detected by MR alone, based on focal enhancement. CONCLUSION: Accordingly, contrast-enhanced MR is not recommended during the first 9 months after therapy. Nine to 18 months after therapy, it may be helpful in those two-thirds of cases where the scar does not enhance. If enhancement takes place (one-third of cases), it may represent either scar or tumor, and in such circumstances, enhanced MR is of no value. After 18 months, enhanced MRI has proven a valuable additional tool. By correctly detecting or excluding recurrent tumor, it can significantly improve diagnostic accuracy. PMID- 8227575 TI - MRI of hepatocellular carcinoma before and after transcatheter chemoembolization. AB - OBJECTIVE: Magnetic resonance imaging of hepatocellular carcinomas (HCCs) was performed before and after transcatheter arterial chemoembolization (TACE). The changes of tumor signal intensity were compared to residual tumor or tumor recurrence in the follow-up period. MATERIALS AND METHODS: Fifteen cirrhotic patients with HCC were studied. All patients were examined with MRI both before and 3 months after TACE. Requirement for the study was that all lesions were detectable by MR before TACE. RESULTS: Magnetic resonance imaging detected 31 tumors. The changes of tumor signal intensity were compared to residual tumor or tumor recurrence in the follow-up period. On T1-weighted (T1W) images before TACE, 16 tumors were hyperintense, 11 were isointense, and 4 were hypointense; on T2W images, 26 tumors were hyperintense, 5 were isointense, and none were hypointense. On T1W images 3 months after TACE, 8 tumors were hyperintense, 18 were isointense, and 9 were hypointense; on T2W images, 11 were hyperintense, 11 were isointense, and 9 were hypointense. All the tumors that became hypointense on T2W images after TACE did not recur on follow-up. All lesions that were still hyperintense on T2W images after TACE showed residual tumor during the follow-up. Among 11 tumors that were isointense on T2W images after TACE, 7 were hyperintense on T2W images before TACE; 6 of these did not recur. Four tumors were isointense on T2W images before and after TACE; 2 of these showed residual tumor in the follow-up. On T2W images after TACE, 15 of 16 HCCs that decreased in signal intensity did not recur on follow-up. On T1W images no correlation was seen between the changes of the signal intensity of the lesion and tumor recurrence in the follow-up; however, 9 of 17 HCCs that did not recur after TACE showed decreased signal intensity. All the tumors (10 of 10) with decreased signal intensity on both T1W and T2W images after TACE did not show tumor recurrence on follow-up. CONCLUSION: The results suggest that MRI is useful in the assessment of the therapeutic effect of TACE in HCC. PMID- 8227576 TI - CT-guided paracoccygeal drainage of pelvic abscesses. AB - OBJECTIVE: Using the transrectal/transvaginal routes for the drainage of pelvic abscesses complicating colorectal surgery (anterior resection or abdominoperineal resection) is not always possible. The conventional transgluteal approach through the greater sciatic foramen, although proven to be a valuable access route, can have complications (mainly local pain). MATERIALS AND METHODS: To avoid these difficulties, a CT-guided paracoccygeal-infragluteal approach was used in the percutaneous drainage of deep pelvic (presacral and ischiorectal) abscesses presenting after colorectal surgery in six patients. RESULTS: Percutaneous drainage through this approach was successful in preventing the need for surgery in all six patients. No complications or recurrences were noted, and catheters were removed an average of 15 days after insertion. CONCLUSION: In comparison with the classical transgluteal approach, the paracoccygeal-infragluteal approach minimizes patient discomfort and minimizes the risk of potential injury to the sciatic plexus or blood vessels. This initial series shows that a CT-guided paracoccygeal-infragluteal approach is well tolerated, safe, and effective for the percutaneous drainage of pelvic abscesses developing after colorectal surgery. PMID- 8227577 TI - Factors influencing long-term in vivo reproducibility of QCT overtebral densitometry). AB - OBJECTIVE: We investigated the long-term in vivo reproducibility of quantitative CT (QCT) examinations that were conducted in conformity with a standard and well established methodology. MATERIALS AND METHODS: The long-term reproducibility in vivo of QCT vertebral densitometry was studied in 12 normal postmenopausal women (mean age 51 years), who underwent four to five examinations over the same 2 year period. RESULTS: One group of six patients demonstrated good reproducibility with a coefficient of variation (CV) of bone mineral density (BMD) of < 2.4% and were considered "good cases." The other group of six patients showed poor reproducibility with a CV of BMD of > 3.2% and were considered "problem cases." A statistical study of the technical parameters of the QCT examination was performed to determine and correct for the factors that are correlated with reproducibility errors. Analyzing uncalibrated variations in Hounsfield numbers, we found that the surrounding soft tissues such as muscle, aorta, liver, and fat were correlated in both the good and the problem cases. However, only in the good cases did we find that the Hounsfield numbers of the vertebral bodies correlated strongly with the soft tissues and with parameters characteristic of the calibration regression line. CONCLUSION: Essentially, the calibration procedure appeared to fail in some examinations of the problem cases, causing poor long term reproducibility. This calibration failure may be related to positioning of the subject in the gantry and to variations in electrical parameters of the X-ray tube. A CV > 1% between the four slopes of calibration for the individual vertebrae measured at the same visit may indicate calibration problems and may suggest the need for repeat scanning. PMID- 8227578 TI - MRA in the evaluation of EC-IC bypass patency. AB - OBJECTIVE: Our goal was to determine the value and limitations of three dimensional (3D) time-of-flight (TOF) MR angiography (MRA) for evaluating the patency of extracranial (EC)-intracranial (IC) bypasses. MATERIALS AND METHODS: The 3D TOF MRA was performed in 35 patients to evaluate their EC-IC bypasses (50 sites) at 1.5 T. Subtraction (rephased-dephased) MRA was performed to visualize slowly flowing blood in all patients. Angiographic images were obtained using maximum intensity projection from subtraction images and from rephased images. RESULTS: The patency of bypasses was confirmed at 22 sites and was suggestive at 17 other sites using MRA alone. The diagnostic value of subtraction MRA proved to be superior to that of rephased MRA, and the findings of subtraction MRA correlated relatively well with those of conventional angiography. Edited reprojection MR angiograms were helpful in visualizing collateral vessels without overlapping of vessels. CONCLUSION: Although there have been some problems with MRA so far, it is a useful imaging modality for following up patients who have undergone EC-IC bypass operations, in conjunction with clinical examination, transcranial Doppler sonography, and cerebral blood flow studies using single photon emission CT or PET. PMID- 8227579 TI - Apparent vascular occlusion on cranial TOF MRA with peripheral presaturation technique. AB - OBJECTIVE: Improvement in the visibility and accurate representation of intracranial vessels may be achieved on cranial time-of-flight (TOF) MR angiography (MRA) when the signal intensity of orbital and superficial fat is suppressed. This may be accomplished by placement of spatial radiofrequency presaturation bands along the periphery of the head. The objective of this study was to evaluate the potential for this peripheral presaturation technique to result in a spurious appearance of vascular occlusion. MATERIALS AND METHODS: Three-dimensional TOF MRA of the head was performed in three patients. Flow compensated spoiled gradient echo images were acquired in the transaxial plane, using the peripheral presaturation technique. The MRA acquisitions were then subsequently repeated in each patient following retraction of the peripheral presaturation bands. A phantom study was also performed to assess the slice profile of the presaturation bands. RESULTS: There was absence of flow-related enhancement, indicating apparent vascular occlusion, involving one carotid siphon in each patient on initial MRA images. Following lateral retraction and/or removal of presaturation bands, the carotid siphons were documented to be patent, with normal caliber and flow-related enhancement demonstrated. Image analysis and phantom experiments indicate this vascular pseudoocclusion is due to proximity of the lateral presaturation band to the lateral aspect of the petrous segment of the internal carotid artery. In this location, there is inadvertent saturation of flowing spins due to imperfections in the presaturation band slice profile. CONCLUSION: The use of peripheral presaturation bands for cranial TOF MRA is useful for improving vascular depiction. However, patent vessels may occasionally demonstrate an absent signal intensity, suggesting occlusion when this technique is used. Awareness of this diagnostic pitfall is important so that serious misdiagnosis does not occur. PMID- 8227580 TI - MRI of intraventricular cysticercosis: surgical implications. AB - OBJECTIVE: Our goal was to evaluate the role of Gd-enhanced MR in the diagnosis and surgical planning of intraventricular cysticercosis cysts. MATERIALS AND METHODS: Thirty-three patients with intraventricular cysticercosis were evaluated with Gd-enhanced MRI including follow-up studies ranging over time periods from 6 months to 8 years. The patient age ranged from 17 to 65 years. All had lived in Mexico or Central America. RESULTS: Twenty patients had surgical removal of the cysts. The majority of patients (18 of 20) who had surgical removal of the cyst showed improvement with resolution of hydrocephalus. Two of 20 patients required additional shunting to alleviate hydrocephalus. In these two patients, Gd enhanced MR showed cyst wall enhancement. Ependymitis with adhesion was found at surgery in these two patients. Nine patients had shunt placement; one of nine patients showed cyst enlargement 2.5 years later that required surgical removal. Of the remaining eight patients, four showed irregular, thick, ring-like enhancement mimicking a neoplasm on follow-up imaging studies. Four patients with cysts in the lateral ventricle did not require treatment. Two of the four patients showed irregular, thick, ring-like enhancement on follow-up imaging studies. CONCLUSION: Gadolinium-enhanced MR is more sensitive than contrast enhanced CT for detecting ependymitis, which is essential in deciding whether surgical removal of the cyst or shunt placement is indicated. A degenerating intraventricular cysticercosis cyst may present as an irregular, thick, ring-like enhancing lesion or a nodular enhancing lesion with varying degrees of surrounding edema, mimicking a neoplasm. PMID- 8227581 TI - Gd-DTPA enhancement of CSF in meningeal carcinomatosis. AB - OBJECTIVE: We report four cases of CSF enhancement secondary to meningeal carcinomatosis observed during MRI. Only one case has been reported previously. MATERIALS AND METHODS: Four patients ranging from 4 to 20 years of age, and all having primary or secondary CNS neoplasms, were examined by cranial and/or spinal MRI before and after Gd-DTPA administration. Three of the patients had additional delayed imaging, and subtraction was used in one case. RESULTS: All four patients demonstrated CSF enhancement after Gd-DTPA administration, which increased on delayed imaging and was more apparent following subtraction. Three of the four patients died within 5 months of the MRI examination. CONCLUSION: CSF enhancement is uncommon, but when seen indicates massive tumor that coats the surface of the CNS. Detection of CNS enhancement may alter therapy; however prognosis may be poor when CSF enhancement is present. Delayed imaging and subtraction may improve detection of CSF enhancement. PMID- 8227582 TI - Influence of temperature on QCT: implications for mineral densitometry. AB - OBJECTIVE: Inaccuracies in quantitative CT (QCT) for vertebral bone mineral measurements may result from differences between the temperature of the vertebrae and the calibration standards. This study aims to quantify these effects by using scans of marrow-equivalent materials and computer simulation. MATERIALS AND METHODS: The CT numbers of fat, water, gelatin suspension, and bone were measured within an anthropomorphic phantom at temperatures between -18 and 38 degrees C. A computer simulation was then performed using these materials to represent marrow fat, soft tissue, and bone in varying proportions over this temperature range. Postprocessing single and dual energy QCT calculations were then performed on the data acquired from the simulation. RESULTS: A change of 80 HU in the CT number of water on cooling from 38 to -18 degrees C was demonstrated. An increase of 95 HU in the CT number of fat occurred over the same temperature range. Dry cortical bone showed no change in CT number with temperature changes from 24 to -18 degrees C. In the computer simulation, the fat error associated with single energy QCT for trabecular bone mineral densitometry was 20% less for specimens at room temperature than at body temperature. In simulated frozen specimens, varying marrow fat/soft tissue composition had almost no effect on single energy QCT mineral densitometry. Dual energy QCT methods that use a fat-equivalent reference material were significantly influenced by the temperature of the specimen. CONCLUSION: The fat error of single energy QCT for mineral densitometry may have been underestimated in previous in vitro studies using vertebral specimens scanned at room temperature. In the simulation, the fat error diminished as the temperature of the specimen was reduced and was negligible when frozen. Fat equivalent reference materials used for dual energy QCT in vivo should have similar X-ray-attenuating properties at room temperature to those of marrow fat at body temperature. PMID- 8227583 TI - Computer-assisted coregistration of multislice SPECT and MR brain images by fixed external fiducials. AB - We have developed and validated in a phantom a method of computer-assisted coregistration using multislice SPECT and MR images. Reusable fiducial markers were fabricated from nylon-based plastic and consist of two parts: a base that remains fixed to the skin with adhesive between scans and a removable, spherical cavity insert that can be filled with contrast agents appropriate for multiple imaging modalities. Markers external and internal to a three-dimensional brain phantom provided a means of quantifying the method's accuracy. A computer algorithm was used to derive transformation matrices for image sets by minimizing the root mean squared deviations obtained for multiple permutations (n = 10) of increasing numbers (range 3-11) of external SPECT/MRI point pairs. As defined by the average +/- SEM mean residual deviations of noncoregistered internal fiducials, the minimal accuracy was 2.4 +/- 0.3 mm (no marker > 3.6 mm) for three coregistration points and did not improve beyond seven to eight fiducials (2.2 +/ 0.1 mm; no marker > 3.2 mm). The method's true accuracy is likely to be better than estimates of minimal accuracy, however, since such measures reflect surmountable random errors in fiducial location. With use of identical MRI (or SPECT) data sets, measures of intraoperator (0.5 +/- 0.1 and 0.5 +/- 0.1 mm, respectively) and interoperator (0.5 +/- 0.3 and 0.8 +/- 0.1 mm) reliability were also obtained, establishing the method as highly reproducible and objective. Preliminary results in a human subject suggest its feasibility for clinical studies. PMID- 8227584 TI - Hallervorden-Spatz disease with bilateral involvement of globus pallidus and substantia nigra: MR demonstration. AB - Hallervorden-Spatz disease presents with relentless mental and physical deterioration. Pathologically there is abnormal iron deposition in the globi pallidi, substantia nigra, and red nuclei. A spectrum of abnormal basal ganglia signal intensities have been described on MRI. A pediatric case with low signal intensity in the globi pallidi and substantia nigra is presented. PMID- 8227585 TI - Prolactinoma invasion of superior ophthalmic vein: CT and MR findings. AB - We report a case in a 6-year-old boy of orbital invasion by prolactinoma in which the cavernous sinus was involved with direct spread into the superior ophthalmic vein, as seen with CT and MRI. PMID- 8227586 TI - Paraganglioma of the filum: MR appearance. AB - Paragangliomas are benign tumors that are thought to arise from the neuroectodermally derived paraganglionic cells of the sympathetic nervous system. We report the imaging findings of an unusual case of paraganglioma of the filum terminale. PMID- 8227587 TI - MRI of iatrogenic spinal epidermoid tumor. AB - A child who had received multiple lumbar punctures 6 years previously was found to have an acquired epidermoid tumor of the lumbar spinal canal, a rare intraspinal lesion. The isointense epidermoid tumor showed ring enhancement only after gadolinium administration. PMID- 8227588 TI - Perineural tumor extension of facial malignant melanoma: CT and MRI. AB - A case of retrograde perineural tumor that spread from malignant melanoma of the face is reported. Contrast-enhanced CT and plain and contrast-enhanced MR studies, including fat suppression technique, showed tumor extension along the maxillary division of the trigeminal nerve. Histopathological examination of the surgical specimen revealed neurotropic malignant melanoma and confirmed the extent of tumor spread in the perineurium. PMID- 8227589 TI - Parathyroid adenoma localization by PET FDG. AB - A patient with primary hyperparathyroidism was examined with positron emission tomography using [18F]-2-fluorodeoxyglucose. The radioactive tracer accumulated in a cervical mass that proved to be a parathyroid adenoma resulting in the correct preoperative localization of the parathyroid tumor. PMID- 8227590 TI - Primary cardiac lymphoma: CT and MR findings. AB - Primary cardiac lymphomas, defined as those involving only the heart and pericardium, are extremely rare tumors. These tumors are commonly fatal and until recently were rarely diagnosed antemortem. We describe the CT and MR findings in a case of primary lymphoma of the heart currently responding to chemotherapy. PMID- 8227591 TI - SVC syndrome as a late complication of ascending aortic aneurysm repair: MR diagnosis. AB - Imaging evaluation is important in the follow-up of patients who have undergone surgical repair of the aorta. We present a case in which MR imaging demonstrated compression of the superior vena cava (SVC) as a late complication of ascending aortic aneurysm surgery. This complication led to thrombosis and clinical SVC syndrome. PMID- 8227592 TI - Skeletal muscle infarction in diabetes: MR findings. AB - We present a case of multiple skeletal muscle infarctions in the lower extremities of a diabetic. Magnetic resonance findings and differential diagnosis are discussed. PMID- 8227593 TI - MR and CT findings in cerebral sparganosis. PMID- 8227594 TI - Intranasal midazolam for claustrophobia in MRI. AB - The authors present their preliminary results using intranasal midazolam for claustrophobic MRI patients. This route of administration reduced the necessity for intravenous sedation from 67 to 17% in this select group of patients. The only side effect encountered was a burning sensation of the nasal mucosa. Further investigation is necessary to determine the efficacy of intranasal midazolam in claustrophobic patients scheduled for MR examinations. PMID- 8227595 TI - PC-based multiparameter full-color display for tissue segmentation in MRI of adnexal masses. AB - OBJECTIVE: Our purpose was to apply full-color composite generation methods to multiparameter MRI to assess the ability of the technique to quantitatively segment clinically important anatomic and pathologic tissues. MATERIALS AND METHODS: With use of a personal computer with a 386 microprocessor and full-color (24 bit) graphics display capabilities, custom and commercially available image processing softwares were applied to spatially aligned multiparameter SE MR image sets obtained from six patients undergoing diagnostic work-up for suspected adnexal or pelvic masses to generate intensity-based color composites. To quantitatively assess the ability of this technique to differentially segment anatomically and pathologically confirmed tissue types into unique color regions within the full-color spectrum, color image analysis was performed on the multiparameter color composites within each patient case, and the results were compared using 95% confidence intervals. RESULTS: Based on the results of pathologic correlation and color image analysis, the generation of full-color composites represents a feasible technique for compressing the diverse tissue contrast data present in multiparameter MR images of adnexal masses. CONCLUSION: With this technique, it is possible to generate composites that simultaneously display uniquely color-coded anatomic and pathologic tissue information within the context of partially natural-appearing images. PMID- 8227596 TI - Formative evaluation of a nursing orientation program: self-paced vs. lecture discussion. AB - Formative program evaluation is a useful process to assess the effectiveness and stability of changes in a program over time. This process documented the effectiveness of changing a medical center nursing orientation program from traditional lecture-discussion to self-learning modules (SLMs). The results of the formative program evaluation confirmed that SLMs were the method of choice for most learners. SLMs focus on adult learning principles more than traditional methods, i.e., lecture-discussion. However, the results of this program evaluation also indicate that staff development educators need to vary teaching methods along with the use of SLMs. PMID- 8227597 TI - Creating bicultural experiences in nursing. AB - This article describes the process (activities) involved in helping Registered Nurse students from Pakistan in an international health project adjust to Canadian culture and readjust to their home culture. The process, involving both structured and informal activities in Pakistan and in Canada, was designed to assist the students in adapting to both the foreign and home cultures. These processes included both human and material resources. Predeparture and reentry workshops, support systems in the form of Karachi-based faculty advisers, and intensive orientation programs were identified as important factors in the project students' adjustment. PMID- 8227598 TI - Developing a community-wide HIV/AIDS nurse education series: a strategy for success. AB - The need for continuing nursing education in the area of HIV/AIDS care and prevention has intensified. Planning community-wide HIV/AIDS programs is a challenge. This article describes a strategy that was successful in consolidating multi-agency expertise and in establishing a framework for subsequent continuing education programs. PMID- 8227599 TI - Mentorship: Maslow and me. AB - The Department of Veterans Affairs Nursing Service developed a leadership training program that utilizes a preceptor for a trainee. The relationship these two establish determines the level of development the trainee can achieve. The working relationship between preceptor and trainee in this setting surpassed the task-oriented aspects of the preceptorship program and can better be described as that achieved through mentorship. The cadre of competencies are comparable to Maslow's (1970) hierarchy of needs and are viewed as interacting and cyclical. The goal of the training program is to develop a leader that is capable of self actualization in an advanced leadership role. The authors' personal experiences are used to demonstrate how the principles of mentorship are applied in the training setting. PMID- 8227600 TI - Meeting the continuing education needs of pediatric nurses: the Pediatric Nurse Exchange Program. AB - Today's dynamic health care environment offers multiple challenges as hospitals attempt to design and implement successful nursing career plans and support the development of professional nurses along such plans. The Pediatric Nurse Exchange Program is designed as a means for personal and professional development to support professional nurses as they are challenged to provide quality nursing care to children and their families. A pilot project is designed to provide experiences for professional nurses in three children's hospitals in Ohio. Exchange experiences, of two to five days in length, provide opportunities for nurses to share expertise, exchange patient care strategies, and develop skills as consultants, change agents, educators, and communicators. Nurse participants in the program are awarded continuing education contact hours for the experience. PMID- 8227601 TI - Using a newsletter to deliver the message of cost containment. AB - Nurse educators often ask themselves how they can affect the bottom line of their organization. At the University of Minnesota Hospital and Clinic, Education Services is using a newsletter to promote cost-effective practice. "Dollars and Sense" has proven to be an effective and efficient educational methodology. A small investment of time and resources can reap substantial savings at the bedside. This article discusses how to develop a similar newsletter. PMID- 8227602 TI - Preceptorship policy: a tool for success. AB - A preceptor policy can form the basis for all elements of a successful program. This article identifies the essential components of a nurse preceptorship program and the responsibilities of the nurse preceptor, preceptee, nurse manager, and staff development educator. Guidelines are included for selecting a nurse preceptor. PMID- 8227604 TI - Creating an effective flyer. PMID- 8227603 TI - Introductions: necessary formality or poetic prelude? AB - Whether we are introducing a program, a speaker, or an issue, our purpose is to convey information, spark interest, and create an attitude, preferably a positive one. Many of us tackle the challenge of introductions lightly, exerting no more time and energy than minimally necessary. Is this lack of preparation detrimental to our purpose? This article suggests that introductions can significantly affect the success of a gathering. Specific ways to enrich an introduction are presented. PMID- 8227605 TI - Examination of cutaneous T-cell lymphoma for human herpesviruses by using the polymerase chain reaction. AB - The etiology of cutaneous T-cell lymphoma remains unknown, although an association with viral infection, in particular certain retroviruses and human herpesviruses, has been suggested. The purpose of this study was to examine skin biopsies of cutaneous T-cell lymphoma for the presence of Epstein-Barr virus, herpes simplex virus type 1 and type 2, and human herpesvirus-6 by using the polymerase chain reaction. Lesional skin biopsies from 30 patients with cutaneous T-cell lymphoma were studied. Control specimens included biopsies from 9 patients with lymphomatoid papulosis and 10 patients with pityriasis lichenoides et varioliformis acuta. DNA extracted from each specimen, as well as from a known positive control for each virus, was examined by using the polymerase chain reaction with viral-specific primers. Each DNA specimen was also amplified with control primers for human beta globin. The specificity of the amplified products was confirmed by Southern analysis. Neither Epstein-Barr virus nor herpes simplex virus was detected in any of the patient specimens examined. Human herpesvirus-6 was detected in one specimen of cutaneous T-cell lymphoma and one specimen of lymphomatoid papulosis. These results do not support a role for any of these herpesviruses in the pathogenesis of cutaneous T-cell lymphoma. PMID- 8227606 TI - Clear cell basal cell carcinoma: an unusual degenerative variant. AB - Clear cell basal cell carcinoma (BCC) is an unusual variant of BCC which is characterized by a variable component of large clear cells. Twenty cases are presented which are further subdivided into pure, mixed, or metatypical categories. Although initially thought to represent sebaceous or tricholemmal differentiation, electron microscopic studies demonstrated that the clear cell changes are a degenerative phenomenon, probably involving lysosomes. PMID- 8227607 TI - Antibody deposits in Tzanck smears in pemphigus vulgaris. AB - Forty-three patients, including 24 males and 19 females between 5 and 62 years of age, having pemphigus vulgaris (27), pemphigus foliaceus (1), bullous pemphigoid (3), chronic benign bullous dermatosis of childhood (2) and herpes zoster (10) were included in this study. Tzanck smears were prepared from the floor of the blisters in these patients by deroofing the bullae, and the slides were stored without fixation at room temperature for 1 to 10 days. Immunofluorescence staining was done with FITC-conjugated anti-human IgG. Twenty-one cases having pemphigus vulgaris and 1 case having pemphigus foliaceus showed bright green fluorescence on the membrane of acantholytic cells. No epithelial cells were seen in smears from bullous pemphigoid and chronic benign bullous dermatosis of childhood, whereas epithelial cells were seen in 10 cases of herpes zoster. These stained negative with anti-IgG. Storage of the prepared smears for 1-10 days did not seem to affect the results of immunofluorescence. Tzanck smears can be used as an easy substitute for skin/mucosal biopsy for the direct immunofluorescence test. PMID- 8227608 TI - Post-kala-azar dermal leishmaniasis: a light and electron microscopic study of 18 cases. AB - Post-kala-azar dermal leishmaniasis (PKDL) is caused by the protozoan parasite, Leishmania donovani, and is seen in patients with history of having been treated earlier for the visceral disease form, kala-azar, caused by the same organism. The findings from 18 patients with PKDL are described in this study. The skin manifestations ranged from hypopigmented macules to infiltrated plaques and nodules. Histopathologic examination revealed a cellular infiltrate of lymphocytes, plasma cells, and macrophages. The macrophages were scattered amidst the infiltrate without any localization. In hypopigmented lesions, the infiltrate was confined to the perivascular region in the superficial dermis and was composed mainly of lymphocytes and few plasma cells. In the nodular lesions, the infiltrate occupied the entire thickness of the dermis. Leishman-Donovan bodies were scarce and identified in 16 cases after a prolonged search of Weigert's iron hematoxylin-stained sections. In 2 cases, Leishman-Donovan bodies were not demonstrable. Electronmicroscopic study revealed parasitized macrophages which showed no structural evidence of activation despite the active cellular response around them. The fine structure of the parasites in the histiocytes was also well maintained. This unusual tropical dermatosis is a unique example of change in organotropism of a parasite associated with a change in the host response. PMID- 8227609 TI - Aluminum tattoo: a phenomenon that can resemble parasitized histiocytes. AB - We present a study of aluminum tattoos that resulted from aluminum chloride cauterization of biopsy sites. The stippled macrophages were studied by light and electron microscopy and by means of X-ray energy spectroscopy (XES) analysis. The light microscopic appearance may be striking, and may mimic the appearance of parasitized histiocytes, although the particles are larger and vary in size and shape. The basophilic particles are electron dense by electron microscopy. XES analysis demonstrated distinct emission peaks corresponding to aluminum, calcium, and phosphorus. Only a minor chloride emission peak was identified. The particles may represent an inorganic precipitate or an organic compound in which aluminum, calcium, and phosphate form a complex with protein. PMID- 8227610 TI - Quantitative study of the human hair follicle in normal scalp and androgenetic alopecia. AB - We carried out a quantitative study of the matrix and dermal papilla of the human hair follicle of the scalp, both normal and in various degrees of androgenetic alopecia. A stereological study showed the measured parameters to decrease with increase in the degree of alopecia, particularly as regards the total volume of the matrix and its papilla. The ratio of the two volumes increased by 30%, indicating a much more marked decrease in size of the papilla than in the matrix size. The number of cell nuclei in the matrix and the papilla of alopecic scalp was found to be 30% and 50% smaller, respectively, than those of normal scalp. Finally, a morphometric study revealed enlarged nuclei in the matrix and papilla, as shown by their increased areas, perimeters, and maximal diameters. PMID- 8227611 TI - Acquired perforating dermatosis of diabetes mellitus and renal failure: further ultrastructural clues to its pathogenesis. AB - An ultrastructural study of a typical case of acquired perforating dermatosis in a patient with renal failure and diabetes mellitus is reported. Crystal-like microdeposits of an electron-lucid material were detected in the upper dermis, close to the transepidermal channel. Compact macrophage conglomerations surrounded the deposits, and a strong histiocytic response was present. Mononuclear inflammatory cells of "activated" type penetrated the acanthotic epidermis provoking basement membrane dissolution and widening of interkeratinocyte spaces. Collagen fibers were seen in the keratotic plug, indicating the process of transepidermal elimination. Our observation supports the hypothesis suggesting that some kind of storage phenomenon may be at the origin of perforating skin lesions in renal failure patients. PMID- 8227612 TI - Intravenous glomus tumor of the forearm. AB - An intravenous glomus tumor occurring in a forearm vein is reported. The patient had a painful subcutaneous mass which was completely excised. This mass was a neoplasm which expanded the lumen of a vein and extended throughout its wall into the surrounding subcutaneous fat. The neoplasm consisted of sheets of rounded cells with a capillary stroma. The neoplastic cells were closely apposed to the capillary vessels and were positive for vimentin, smooth muscle actin and muscle specific actin. The cells were negative for desmin, factor VIII-related antigen, epithelial membrane antigen, cytokeratins, S-100 protein and chromogranin. This is the 2nd reported case of intravenous glomus tumor of the forearm. This unusual presentation may be due to intravascular extension by a cutaneous glomus tumor. The potential for intravascular growth by glomus tumor should be recognized by surgeons, dermatologists and pathologists. PMID- 8227613 TI - Localized hair follicle hamartoma. AB - A 4-year-old male presented with an indurated papular plaque on his right nasolabial fold. The lesion, present since the child was a few months of age, progressively extended in a band-like fashion up to the upper lip. Microscopically, there was almost complete replacement of the hair follicles by trichoepithelioma-like epithelial proliferations associated with hyperplastic sebaceous glands. We interpret this lesion as a localized form of the clinicopathological entity known as generalized hair follicle hamartoma. PMID- 8227614 TI - Malignant melanoma arising in a nevus of Nanta. AB - Papillomatous fleshy facial nevi have a low risk of malignant transformation. In addition, they may undergo ossification in their depths; a nevus with bone is known as a nevus of Nanta. Such a nevus in a 60-year-old female gave rise to a malignant melanoma. PMID- 8227615 TI - A unique case of apocrine carcinoma on the male pubic skin: histopathologic and ultrastructural observations. AB - We report a case of cutaneous adenocarcinoma on the pubic region of a 56-year-old male. The exophytic tumor showed solid, ductal, and glandular patterns with decapitation secretion and mucinous carcinoma-like nests. Despite the exophytic nature of the lesion, an invasive growth pattern into the deep dermis and subcutaneous fat was seen. An additional characteristic of this tumor was the presence of Paget's phenomenon. From these findings, we conclude that this tumor is an apocrine adenocarcinoma having an unusual clinical appearance and unique histopathological features which have some similarities to apocrine carcinoma of the breast and some to syringocystadenoma papilliferum and nipple adenomatosis. PMID- 8227616 TI - Risk factors associated with contamination of raw milk by Listeria monocytogenes in dairy farms. AB - A case-control study involving 128 selected dairy farms was conducted to assess the association of several suspected risk factors with the odds of contamination of raw milk by Listeria monocytogenes. Using logistic regression, we found that poor quality of silage (pH > 4.0), inadequate frequency of cleaning the exercise area, poor cow cleanliness, insufficient lighting of milking barns and parlors, and incorrect disinfection of towels between milkings were significantly associated with milk contamination by L. monocytogenes. More attention to preparing silage and good milking and barn hygiene are important for diminishing the risks of exogenous contamination of raw milk by L. monocytogenes. PMID- 8227617 TI - Developmental changes in embryonic resistance to adverse effects of maternal heat stress in cows. AB - The objective of this study was to determine whether bovine embryos become more resistant to deleterious effects of maternal heat stress as early embryonic development progresses. Superovulated, lactating Holstein cows were bred by AI and assigned to be heat stressed on d 1, 3, 5, or 7 of pregnancy (d 0 = day of estrus) or not heat stressed (control). Embryos were retrieved from the uterus on d 8 and evaluated for viability and stage of development. Compared with embryos of control cows, embryos of cows receiving heat stress on d 1 had decreased viability and development. Maternal heat stress on other days had no detrimental effect on embryonic viability or stage of development. Bovine embryos become more resistant to adverse effects of maternal heat stress as pregnancy progresses; substantial resistance develops by d 3. This information may be useful in design of environmental modification systems that provide cooling at critical periods of gestation to enhance pregnancy rates during summer in hot climates. PMID- 8227618 TI - Automated electronic activity measurement for detection of estrus in dairy cattle. AB - Two trials were conducted to characterize the performance of an automated electronic activity tag system as an aid for detection of estrus in dairy cattle. In trial 1, activity tags were attached to the rear leg of 24 cows at approximately 35 d postpartum and remained attached until pregnancy. Data collection included 66 periods of predicted estrus (cyclic periods of 18 to 24 d prior to the date of pregnancy). A summary of the data recorded in the activity tags was transmitted telemetrically to a personal computer at each milking via stationary antenna in the milking parlor. An electronic flag used an increased activity ratio to determine the ratio of activity in a test period of the previous 12 h to activity in the same 12 h during the 2 d previous to the test period, thus indicating estrus. Activity patterns were characterized from data recorded on the tags at 2-h intervals. The tag detected 74% of predicted periods of estrus versus 58% reported by herders. An increased activity ratio for at least 4 consecutive h reduced false-positive designations. In trial 2, activity tags were attached to front and rear legs on five cows, and activity patterns from the two sites were compared for 2 mo. Patterns of activity were similar from tags attached at either site, and the sites were not different in their discrimination between periods in which estrus did or did not occur. The activity tag system was an effective practical tool to detect estrus. PMID- 8227619 TI - Histologic response of the heifer mammary gland to intramammary infusion of interleukin-2 or interferon-gamma. AB - The histologic and cytologic responses of heifer mammary glands infected with Staphylococcus aureus were studied after infusion of interleukin-2 or interferon gamma. Two groups of 4 heifers each, which were infected experimentally with S. aureus in all 4 mammary quarters, were infused in diagonal quarters with 7.5 x 10(5) units of interleukin-2 or 10(5) units of interferon-gamma; remaining quarters received PBS. Heifers in both trials were slaughtered 14 d after cytokine infusion, and mammary tissues were collected for histological examination. Uninfected quarters from 2 additional heifers were left untreated to compare infected with uninfected tissues for both trials. Morphologic evaluation and leukocyte infiltration scores were performed on tissue sections stained with hematoxylin and eosin, and plasma cells were quantified on sections stained with immunoperoxidase. Infected quarters had lower percentages of alveolar epithelial and luminal areas and higher percentages of stromal area than did uninfected quarters in the interleukin-2 trial, but no differences were observed between infected quarters that had been treated with PBS or interleukin-2. Likewise, interferon-gamma treatment had no effect on mammary parenchymal components in the infected quarters. Interleukin-2 treatment significantly elevated leukocytosis into the mammary gland parenchyma compared with infected quarters treated with PBS and uninfected quarters. Among the leukocyte types evaluated, eosinophilic infiltration was elevated in interleukin-2 quarters over that of PBS controls. In both trials, concentrations of plasma cells bearing Ig were elevated in infected versus uninfected quarters. Plasma cell concentrations also were higher in cytokine than PBS controls, especially in interleukin-2 quarters. Results suggested that neither cytokine influenced the histology of infected mammary tissues, but both interleukin-2 and interferon-gamma increased, although insignificantly, the prevalence of all isotypes of plasma cells bearing Ig, suggesting enhancement of the local immune response to IMI. PMID- 8227620 TI - A randomized blind trial on dry cow antibiotic infusion in a low somatic cell count herd. AB - The objective of the study was to evaluate the effect of administration of antibiotics to cows at dry-off in a low SCC herd. Cows were infused at dry-off with an intramammary antibiotic on the left or the right side of the udder. Selection of the infused quarters was random, and all people evaluating the occurrence of mastitis were unaware of which half of the udder was infused. The quarters that were infused with an antibiotic showed a significantly lower incidence of clinical mastitis in the dry period. Quarters that were uninfused had 10 cases of clinical mastitis versus 1 case in quarters that were infused with antibiotics. The quarters that were infused showed a reduction of minor pathogens at calving. The uninfused quarters showed little evidence of spontaneous cure during the dry period. Milk leaking was frequent during the drying off period. Milk leaking was also associated with the occurrence of clinical mastitis later in the dry period. Cows that leaked milk had a four times greater risk of clinical mastitis in the dry period. PMID- 8227621 TI - Effect of prepartum propylene glycol administration on periparturient fatty liver in dairy cows. AB - Plasma glucose concentration during late gestation was thought to be important for the development of fatty liver near parturition. Thirteen multiparous cows were given a 1-L oral drench of propylene glycol once daily beginning 10 +/- 3.6 d prepartum until parturition. Eleven control cows received a 1-L water drench. Plasma glucose increased following propylene glycol administration. Plasma NEFA concentration was 403 and 234 microM, and plasma insulin concentrations were .354 and .679 ng/ml, for control cows and cows treated with propylene glycol measured from 1 to 7 d prepartum. Plasma NEFA tended to be lower in cows treated with propylene glycol from 1 to 21 d postpartum. Prepartum propylene glycol administration reduced hepatic triglyceride accumulation by 32 and 42% at 1 and 21 d postpartum, respectively. Prepartum plasma BHBA was reduced during propylene glycol administration. Prepartum plasma glucose, NEFA, BHBA, and insulin were strongly correlated with liver triglyceride at 1 d postpartum (r = -.49, .45, .36, and -.49, respectively). Pre- and postpartum DMI were not affected by treatment. Milk production and composition measured through 21 d postpartum were not different between groups. PMID- 8227622 TI - Lactation response to ruminally protected methionine and lysine at two amounts of ruminally available nitrogen. AB - Sixty multiparous Holsteins were used in a 200-d continuous lactation trial designed to measure the response to a mixture of ruminally protected Met and Lys. Response to AA was determined at two dietary concentrations of degraded protein designed to provide 85 and 100% of NRC recommended amounts. This difference in degraded protein content was achieved by addition of urea. Diets without urea supported lactation as well as diets with added urea in early lactation. In midlactation, urea addition was detrimental to yields of milk and milk protein; addition of AA prevented this negative effect. In early lactation, AA addition raised milk protein concentration and yield by 1 g/kg of milk of 37 g/d and did not interact with urea. Increases in protein concentration in response to AA addition were similar in early and late lactation and corresponded to changes in the casein fraction of milk. These results support previous work showing the importance of adequate absorbable Lys and Met to maximize the protein content of milk. The results suggest that interactions may occur between ruminal ammonia production and AA supply. PMID- 8227623 TI - Intestinal digestibility in dairy cows of protein from grass and clover measured with mobile nylon bag and other methods. AB - Ruminal, intestinal, and total tract digestibilities of CP in grass, grass silage, and clover were measured using the mobile nylon bag method. Total tract indigestibilities measured with this method were compared with indigestible CP predicted by in vitro, 14-d ruminal incubation, and 12-h followed by 14-d ruminal incubation methods. Indigestible CP was related to chemical composition and harvest date. Ruminal disappearance of CP from nylon bags was 58, 71, and 58% for grass, grass silage, and clover, respectively. Intestinal CP disappearance was 84, 81, and 91% of escape CP, respectively. Lower ruminal disappearance was compensated by higher intestinal disappearance, resulting in a small variation in total tract indigestible CP within forage. Total tract indigestible CP was 7, 6, and 4%, respectively. Average total tract indigestible CP measured with the mobile nylon bag method (6%) was lower than that estimated by the in vitro (13%), 14-d ruminal incubation (14%), or 12-h ruminal incubation followed by 14-d ruminal incubation (11%) methods. All methods ranked forages similarly, resulting in high correlation coefficients among methods within forage. Total tract indigestible CP of grass, grass silage, and clover was related to CP content and harvest date, DM and ADIN content, and DM and CP contents, respectively. PMID- 8227624 TI - Effects of partial replacement of ryegrass by low protein feeds on rumen fermentation and nitrogen loss by dairy cows. AB - Three rumen fermentation studies in combination with three feeding trials were carried out to investigate the effect of partial replacement of heavily fertilized perennial ryegrass by low protein feedstuffs on pH and concentrations of VFA and NH3 N in the rumen and on N excretion in milk, urine, and feces by dairy cows. Feedstuffs tested were high fiber concentrate mixtures based on sugar beet pulp and soybean hulls and high starch concentrate mixtures based on corn (Experiments 1 and 2), corn silage (Experiment 1), dried and ensiled pressed sugar beet pulp and high moisture ear corn silages with or without husks (Experiment 3). In the fermentation studies, N intake ranged between .43 and .57 kg/d. Partial replacement often increased DMI (maximal by 2.6 kg), resulting in minor effects on N intake. Urinary N excretion ranged between 30 and 58% of N intake and decreased by 30 to 40% when grass was partially replaced. Fecal N output was between 25 and 30% of N intake and tended to increase with the low protein feed. The reduction in urinary N excretion corresponded to a decrease of rumen NH3 N. Replacement by concentrate mixtures based on corn reduced milk fat content; for mixtures based on beet pulp, milk fat content was not changed. PMID- 8227625 TI - Comparison of copper lysine and copper sulfate as copper sources for ruminants using in vitro methods. AB - Two in vitro experiments were conducted to estimate the availability of Cu from Cu Lys compared with Cu from CuSO4. In experiment 1, a 24-h ruminal fermentation (alpha-cellulose substrate) containing either .1% added S or 2% added urea was performed with 4, 12, or 96 ppm of added Cu as either Cu Lys or CuSO4. Soluble Cu was measured at the end of the 24-h incubation, and concentrations were higher for Cu Lys with added urea (1.16 vs. .45 ppm), but no differences existed between sources of added S. In Experiment 2, 20 ppm of Cu added as either Cu Lys or CuSO4 with and without the addition of 10 ppm of Mo and .75% S were used in an vitro study designed to simulate digesta passage through the ruminant. A 24-h ruminal fermentation (orchardgrass substrate) was followed by a 2-h digestion in pepsin and HCl (pH 2.3) and then a 2-h digestion in NaHCO3 and pancreatin (pH 6.6). Soluble Cu concentrations were analyzed after each step. Molybdenum and S addition decreased soluble Cu after ruminal fermentation and tended to decrease soluble Cu concentrations after digestion of pancreatin and NaHCO3. The source of Cu did not affect soluble Cu concentrations. Results suggest that Cu from Cu Lys and CuSO4 behave similarly in the digestive tract of ruminants. PMID- 8227626 TI - Mineral accretion in the fetus and adnexa during late gestation in Holstein cows. AB - Multiparous Holstein cows (n = 18) were bred artificially to the same Holstein bull and then slaughtered at times ranging from 190 to 270 postmating to assess mineral accretion by the conceptus. Fresh weight, DM, and concentrations of Ca, P, Mg, K, Na, Fe, Zn, Cu, and Mn were obtained for the fetus, fetal fluids, fetal membranes, cotyledons, caruncles, and uterine tissues. Rates of accumulation of individual minerals in different components of the conceptus during late gestation were described by either linear or exponential relationships. Estimated Ca accretion rate in the conceptus increased from 2.3 g/d at 190 d of gestation to 10.3 g/d at 280 d of pregnancy; corresponding P accretions were 1.9 and 5.4 g/d. Rates of accretion of Mg, K, and Na in the conceptus in late pregnancy were about .2, 1.0, and 1.4 g/d, respectively, and Fe, Zn, Cu, and Mn accumulated in the conceptus at rates of 18.0, 11.7, 1.6, and .3 mg/d, respectively. These daily rates represent net mineral requirements for conceptus growth during late pregnancy in mature Holstein cows. In general, our values are consistent with current dietary recommendations for minerals during the dry period. PMID- 8227627 TI - Effect of infused volatile fatty acids and caseinate on milk composition and coagulation in dairy cows. AB - Milk protein secretion is changed by increasing the proportion of energy, mainly as propionic acid, or the availability of AA. Whether associative effects exist between energy nature and protein amounts is unknown. Therefore, ruminal isoenergetic infusions of low or high propionate mixtures were combined factorially with duodenal infusion of sodium caseinate or control. Four ruminally and duodenally fistulated Holstein cows were used. The diet was limited and consisted of 70% forage and 30% concentrate. Caseinate infusion increased milk yield and protein and casein contents and decreased milk fat content; curd yields and coagulation properties of milk were improved. The infusion of propionic acid caused a large increase in rumen propionate. Milk yield tended to decrease, and milk fat decreased, but protein, casein, and curd yields were unchanged; milk coagulating properties were improved. No interaction existed between energy and protein amounts. Alteration of VFA had little effect on milk composition, but increasing the protein supply to the duodenum increased milk protein. PMID- 8227628 TI - Effect of fructose-induced hypoglycemia on cessation of macromolecular transport in the neonatal calf. AB - Fructose was used as a hypoglycemic agent to determine whether availability of glucose during early life mediates the cessation of intestinal transport of Ig (closure). Either glucose or fructose (100 g in 2 L of electrolyte solution) was fed to calves deprived of colostrum at 3, 6, 9, 12, 15, 18, and 21 h postpartum. Colostrum (1 kg) was fed to all calves at 24, 36, and 48 h postpartum. Venous blood was sampled, and plasma was harvested, for the measurement of glucose, fructose, insulin, and IgG. During the first 24 h, means for plasma glucose from calves treated with glucose and fructose, respectively, were 282 and 24 mg/dl. The latter value reflects the degree of hypoglycemia induced by feeding fructose. During the same period, corresponding concentrations of plasma fructose were 4 and 230 mg/dl. Plasma insulin concentrations for precolostral calves were 101 and 11 microU/ml for calves treated with glucose and fructose, respectively. Peak IgG concentrations were extremely low for both groups of calves, indicating that intestinal transport of Ig had virtually ceased by the end of the 24-h treatment period. Thus, fructose-induced hypoglycemia had no effect on the time of intestinal closure in the newborn calf. PMID- 8227629 TI - Evaluation of milk antibiotic residue screening tests in cattle with naturally occurring clinical mastitis. AB - Milk from 172 commercial cows with mild to moderate clinical mastitis was tested with five antibiotic residue detection assay systems. One hundred cows were treated with one of two intramammary beta-lactam antibiotics, and the remaining 72 cows were treated with intramuscular oxytocin. Milk samples were collected pretreatment, twice after therapy, and again 21 d following the initiation of treatment. Presumptive false-positive assay results were tabulated from all pretreatment and 21-d milk samples and from samples collected following oxytocin therapy. The percentage of false-positive results was 43.6, 37.7, 81.7, 2.6, and 18.8% for the CITE probe (beta-lactam), Delvotest-P, Charm Farm, LacTek (beta lactam), and Bacillus stearothermophilus var. calidolactis disk assay, respectively. In four of the assay systems, average SCC were significantly higher in samples yielding false-positive results than in those with negative results. Specificity and sensitivity were estimated for each assay system, and, based on these estimates, positive and negative predictive value curves were graphed as the prevalence of milk samples containing detectable concentrations of exogenous antibiotic residues in the sample population was varied from 0 to 100%. PMID- 8227630 TI - Review and update of casein chemistry. AB - Of all food proteins, bovine milk proteins are probably the most well characterized chemically, physically, and genetically. The primary structures are known for most genetic variants of alpha s1-, alpha s2-, beta-, and kappa caseins, beta-lactoglobulin, and alpha-lactalbumin. Secondary and tertiary structures of the whey proteins have been determined, and secondary structures of the caseins have been predicted from spectral studies. The caseins, although less ordered in structure and more flexible than the typical globular whey proteins, have significant amounts of secondary and, probably, tertiary structure. The amphipathic structure of the caseins is especially noteworthy; thus, these proteins most likely are divided into polar and hydrophobic domains. The presence of anionic phosphoseryl residue clusters in the calcium-sensitive casein polar domains is particularly significant because of their interaction with calcium ions, or calcium salts, or both, and the formation of micelles. Flexibility of casein structures is reflected by their susceptibilities to limited proteolysis, which dramatically changes functionality. PMID- 8227631 TI - The secondary structure of milk proteins and their biological function. AB - A brief overview is given of the methods of determining and predicting secondary structure in proteins. The secondary structures of the milk serum proteins, lactoferrin, alpha-lactalbumin, lysozyme, and beta-lactoglobulin, as determined by x-ray crystallography, are compared with the results of a joint prediction method. This comparison evaluates critically the degree of success achieved and helps define what can reasonably be expected from a prediction in the absence of a known structure. The value of supplementary information from spectroscopic methods and the use of templates and sequence information from related proteins in improving the confidence of predictions are illustrated. One point that emerges is the general overprediction of helix content by the joint prediction method such that, for an all-beta protein such as beta-lactoglobulin, the method of Garnier, Osguthorpe, and Robson, applied with the correctly selected decision constants, provides a somewhat better approach. Secondary structure of the caseins can be predicted with less confidence than for globular proteins, and the results should be interpreted as evidence of a propensity to form transient secondary structures of the indicated type in view of the generally open and flexible conformation of caseins in solution. PMID- 8227632 TI - Structure and function of milk protein genes. AB - Interspecies comparisons of cDNA and mosaic milk protein genes have confirmed their high rate of evolution, but the overall gene organization has been conserved. The three Ca-sensitive casein genes, which share common motifs in the promoter region and contain similar sequences that encode signal peptide and multiple phosphorylation sites, probably derived from a common ancestor. alpha s1 and alpha s2-casein genes, divided into many small exons, undergo complex splicing, and the deleted caseins arise from exon skipping. The four bovine casein genes are clustered on 200 kb of chromosome 6. alpha-Lactalbumin and beta lactoglobulin pseudogenes occur in ruminants. Study of the expression of native and modified milk protein genes in mammary cell lines and transgenic animals and DNA footprinting have shown the occurrence of important regulatory motifs in the proximal 5' flanking region, including one recognized by a specific mammary nuclear factor. Good stage- and tissue-specific expression has been obtained in transgenic animals with milk protein genes having less than a 3-kb 5' flanking region. Better knowledge of both the structure and function of milk protein genes, which has already allowed the use of powerful techniques for the rapid identification of alleles, offers the potential for the genetic modification of milk composition. PMID- 8227633 TI - Developing methodology for microbiological evaluation of milk and dairy products- an introduction. PMID- 8227634 TI - Rapid methods and automation in dairy microbiology. AB - The importance of microbiology to the dairy industry has been demonstrated by recent outbreaks of foodborne illness associated with consumption of milk and dairy products that had been contaminated with pathogenic organisms or toxins. Undesirable microorganisms constitute the primary hazard to safety, quality, and wholesomeness of milk and dairy foods. Consequently, increased emphasis has been placed on the microbiological analysis of milk and dairy products designed to evaluate quality and to ensure safety and regulatory compliance. The focus of dairy microbiology, however, remains largely on conventional methods: plate counts, most probable numbers, and dye reduction tests. These methods are slow, tedious, intensive in their requirements for material and labor, and often not suitable for assessing the quality and shelf-life of perishable dairy foods. With the exception of coliforms, Salmonella, and Staphylococcus aureus, isolation and characterization of various organisms occurring in milk and milk products are seldom a part of the routine microbiological analysis in the dairy industry. Recent emphasis on the programs based on HACCP (Hazard Analysis and Critical Control Points) for total quality management in the dairy industry and increased demand for microbiological surveillance of products, process, and environment have led to increased interest in rapid methods and automation in microbiology. Several methods for rapid detection, isolation, enumeration, and characterization of microorganisms are being adapted by the dairy industry. This presentation reviews rapid methods and automation in microbiology for microbiological analysis of milk and dairy products. PMID- 8227635 TI - Biosensors: applications for dairy food industry. AB - Biosensors are defined as indicators of biological compounds that can be as simple as temperature-sensitive paint or as complex as DNA-RNA probes. Food microbiologists are constantly seeking rapid and reliable automated systems for the detection of biological activity. Biosensors provide sensitive, miniaturized systems that can be used to detect unwanted microbial activity or the presence of a biologically active compound, such as glucose or a pesticide. Immunodiagnostics and enzyme biosensors are two of the leading technologies that have had the greatest impact on the food industry. The use of these two systems has reduced the time for detection of pathogens such as Salmonella to 24 h and has provided detection of biological compounds such as cholesterol or chymotrypsin. The continued development of biosensor technology will soon make available "on-line quality control" of food production, which will not only reduce cost of food production but will also provide greater safety and increased food quality. PMID- 8227636 TI - Applications of bioluminescence in the dairy industry. AB - Several applications of ATP bioluminescence of relevance to the dairy industry have been proposed. This paper reviews some of the major benefits of the technology. New developments in bioluminescence research have made the simple, rapid, sensitive detection of foodborne pathogens a distinct possibility. PMID- 8227637 TI - Challenges to dairy cattle management: genetic considerations. AB - Several new technologies will be used to produce continued genetic change in dairy cattle. These technologies are categorized broadly as 1) improved modeling, selection, and evaluation methods; 2) use of new and improved reproductive technologies; 3) new developments in molecular genetics; and 4) new developments in immunogenetics. Improvements in evaluation will continue as computers become faster and have more storage capabilities. Improved mathematical models that more nearly describe the biology of lactation will maximize estimation of genetic differences and reduce residuals. New reproductive technology could allow reduction of generation intervals two- to fivefold compared with present generation intervals and, combined with genetic markers, could markedly accelerate progress. Health problems in dairy cattle are expected to increase as production increases. Thus, selection for decreased incidences of health disorders will be needed, probably by selection of sires with improved general immunocompetence. Research is in the early stages of application of techniques of molecular genetics to animal breeding. Early uses will allow detection and alleviation of genetic defects. Eventually, marker genes that directly affect production and metabolic pathways that also affect production will be subjected to selection. The ability to foresee new and potentially useful techniques will be determined by scientific advancement of areas in which researchers are engaged; thus, accurate prediction far into the future cannot be expected. PMID- 8227638 TI - Nutrient requirements versus supply in the dairy cow: strategies to account for variability. AB - Dairy producers must overcome substantial challenges to achieve milk outputs > 14,000 kg of milk/yr per cow within the next decade. To obtain high productivity, a more complete comprehension of the dynamics of metabolism, nutrient utilization, and nutrient absorption will enable better prediction of the efficiency of utilization of these nutrients. A better understanding of the dynamics of rumen function and a more accurate prediction of nutrient flow from the rumen are necessary. Grouping strategy and group feeding behavior influence cow productivity and farm profitability. Understanding of the variance of individual cow responses to management practice is critical. Feeding system design and management and diet formulation techniques need to be developed that recognize the dynamic nature of cow physiology and the variability in feedstuffs and cow requirements. These concepts need to be integrated into total farm management and require the use of new computer modeling technologies. PMID- 8227639 TI - Raising dairy replacements to meet the needs of the 21st century. AB - Producing high quality replacement heifers at minimum cost will be one of the many challenges facing the dairy farm of the 21st century. Because replacement heifers represent a large portion of the total cost of milk production, dairy farmers will have to meet the replacement needs of their lactating herds at minimum cost to maintain the farm's profitability. To keep the US dairy industry competitive into the 21st century, researchers, extension, industry, and producers must examine the database of research in order to determine how to apply the known information to current production and research efforts. Important research is needed to improve the dairy heifers of the future. This review summarizes research of the past 12 yr related to dairy replacements and incorporates those findings into possible scenarios for future dairy calf and heifer management systems. Additional research is needed to determine how dairy replacement raising systems affect the lifetime productivity and profitability of the dairy cow. PMID- 8227640 TI - Record-keeping systems and control of data flow and information retrieval to manage large high producing herds. AB - Record-keeping systems have provided an essential link that significantly increases milk production. As new technologies are introduced, they are integrated into total management programs that provide for proactive management. Maintenance of data flow, not only for the producer but also for other users, requires increased cooperation among the various sectors. Larger production units demand products that integrate production and economic parameters to plan strategically for maximum profitability. PMID- 8227641 TI - Lactation: historical patterns and potential for manipulation. AB - The advent of biotechnology has made data on undomesticated mammals relevant to dairy science. Such data indicate the potential of lactation for modification, reveal genetic material available for use through bioengineering, help distinguish adaptive features from historical artifacts, and clarify limits on lactational diversity that date from early evolution. Evolutionary analysis indicates that a complex degree of lactation preceded divergence of the extant mammalian lineages during the Mesozoic Era. Although aspects of monotreme lactation appear to be ancestral for extant mammals, the marsupials and eutherians exhibit divergent specializations. Evidence is consistent with the idea that protolacteal glands evolved by combining features of skin gland populations into a new functional complex. Secretions of these ancestral glands may have had antimicrobial properties that protected the eggs or hatchlings and organic components that supplemented offspring nutrition. Following development of highly nutritious milks, evolution produced diversity in milk composition and function, milk output, length of lactation, mammary gland anatomy, and contributions of lactation to offspring nutrition. Certain marsupials are specialized in terms of functional independence and temporal plasticity of mammary tissues. Mammalian diversity indicates that artificial selection and physiological manipulation of domestic artiodactyls has only modestly exploited the potential of mammary glands as a nutritional source for humans. PMID- 8227642 TI - Empirical and theoretical constraints on the evolution of lactation. AB - For 829 mammalian species, data on age at weaning or age at first solid food were analyzed with respect to body mass, phylogeny, habitat, diet, length of gestation, basal metabolism, and neonatal development. The primary influence on lactation length is female mass, but phylogenetic constraints are important. Thus, lactation can be characterized as short (earless seals and baleen whales), long (marsupials, bats, and primates), or average (remaining eutherians). Among average mammals, lagomorphs have short lactations. Lactation may have different functions, evolutionary constraints, and physiological control depending on whether young first eat solid food near weaning or well before it. First solid food eaten near weaning occurs in polytocous species with altricial young; in this case, lactation has a clear energetic role. In contrast, first solid food well before weaning is common for mammals with single, precocial offspring. For these species, the energetic and nutritional constraints on lactation may be less important than the benefits of maintaining contact between mother and young, such as reduced juvenile mortality and increased opportunities for learning social or foraging patterns. Thus, the age at first solid food relative to the age at weaning may indicate the function of lactation within the reproductive biology of a given mammal. Delayed development and implantation alter the timing of energetic investment during gestation, so too, the age at first solid food may alter or reflect the rate of energetic investment during lactation. Thus, the age at first solid food relative to the age at weaning may indicate the function of lactation within the reproductive biology of a given mammal. Testing these hypotheses will require data from diverse species on the nutritional and energetic value of milk before and after first solid food as well as on the mechanics and consequences of nursing or suckling during the course of lactation. PMID- 8227643 TI - The adaptation of milk secretion to the constraints of fasting in bears, seals, and baleen whales. AB - Although lactation is accompanied by increased nutrient demands for milk synthesis, many species of bears, true seals, and baleen whales fast for much or all of lactation. Large body mass in these species confers the advantage of greater stores of fat and protein relative to rates of milk production. Given the constraints on substrate availability during fasting, the milks of fasting mammals are predicted to be low in carbohydrate, protein, and water and to be high in fat. The milks of bears, true seals, and baleen whales conform to this prediction. Mammals that lactate while fasting may lose up to 40% of initial BW. The production of milk entails the export of up to one-third of body fat and 15% of body protein in the dormant black bear and in several seal species, which greatly depletes maternal resources and may represent a physiological threshold, because higher protein and fat outputs have only been measured in species that start feeding. The low K:Na ratio of seal and whale milks and the low Ca:casein and inverse Ca:P ratios in seal milks are unusual and warrant further study. PMID- 8227644 TI - Characterization of the management practices of the top milk producing herds in the country. AB - Out of 128 surveys mailed to dairy farms that had been identified by the nine DHI processing centers as the top milk producing herds in the country, 61 surveys were returned. The predominant forage being fed was corn silage, followed by legume hay and haylage. On average, 6.7 feed additives and 3.5 alternative feeds were fed. Producers reported the following incidences of metabolic disorders: parturient paresis, 7.2%; displaced abomasum, 3.3%; ketosis, 3.7%; and nonspecific downer cow syndrome, 1.1%. Producers scheduled detection of estrus 3.1 times per day. Only 18.3% of the herds used routine synchronization of estrus. Artificial insemination was used on 94.8% of the cows and 88.5% of the heifers. The most important trait in sire selection was PTA for milk. For mastitis control, 75.4% of the producers practiced predipping, 85.2% used individual paper towels, 93.4% practiced postdipping, and 95.1% treated all cows at cessation of milking. Increased profitability and lower costs or higher returns were the primary motivators of these producers. Veterinarians were the most frequent source of information, followed by farm magazines. Satellite delivery of educational programs was the least acceptable delivery method. Extension professionals need to be cognizant of the producers' preferred information sources and delivery systems to ensure accurate, timely, cost effective transfer of technological advances. PMID- 8227645 TI - Interactions of high milk yield and reproductive performance in dairy cows. AB - Correlations between reproductive traits and measures of milk yield indicate that higher yield is associated phenotypically and genetically with reduced reproductive performance in lactating cows. Numerous recent studies have reported that reproductive performance is compromised, primarily through delayed ovarian activity and reduced conception rates, by the demands of high milk yield. However, daily managerial decisions to obtain efficient reproductive performance have considerable impact. Management can offset depression in fertility, because high yielding herds often achieve the fewest days open. Selection for milk yield has increased blood concentrations of somatotropin and prolactin, stimulators of lactation, and decreased insulin, a hormone that is antagonistic to lactation and may be important for normal follicular development. These changes in hormone concentrations promote higher milk yield but may be potentially detrimental to other physiological functions, such as reproduction, if the management is not adequate to meet the metabolic demands of lactation. Timing and magnitude of negative energy balance apparently interact to determine the extent to which negative energy balance alters hypothalamic secretion of GnRH and its effect on gonadotropin secretion and, therefore, ovarian secretion of progesterone, which affects expression of estrus and support of the uterus during early pregnancy. PMID- 8227646 TI - New technologies and decision making in high producing herds. AB - New technologies have evolved for improved management of on-farm records and their use in decision making. Computerized on-farm record systems, coupled with automated monitoring of individual animal performance and electronic transfer of data between the on-farm database and mainframe systems, have enhanced the capability for herd management. Electronic aids for decision making may be embedded into on-farm management programs to allow more detailed data to be available with ease and convenience. Technological advances and their application toward improved operational decisions concerning production monitoring, reproductive management, genetic improvement, feeding, and health in a 300-cow dairy herd are presented as examples. Substantial advances have been made in technologies that improve management and decision making. Evolving technologies promise to enhance further the capabilities for improved management and decision making in high producing herds. PMID- 8227647 TI - Optimal genetic improvement for the high producing herd. AB - Genetic merit for milk production is increasing over 150 kg of milk/yr in the Holstein cow population. Nationwide decreases in payments for fat differential since 1987 have not been counter-balanced by similar increases in payments for protein differential, resulting in a steady increase in the value of carrier. Protein yield should be emphasized rather than specific casein or whey fractions of protein because national recording, evaluation, and payment structures are not in place. Genetic merit of bulls for markers may differ across families, making selection for marker traits of limited value in current breeding schemes. Conformation traits that appear to have the highest correlation with measures of herd life are udder depth, for udder attachment, and teat placement. Selection for health traits is difficult because of limited genetic information. National evaluations for SCC are being developed, but average difference between extreme bulls is only slightly more than one linear SCC. Managers of high producing herds should continue with primary selection emphasis on production traits. The greatest challenge to dairy producers is to develop management systems, particularly in nutrition, that allow maximal expression of genetic potential. PMID- 8227648 TI - Developing nutrition programs for high producing dairy herds. AB - Herd average milk production continues to increase in the US. Average milk production in Holstein herds enrolled in DHI testing programs surpassed 9000 kg in some states in 1991. Individual dairy herds have produced > 14,000 kg per cow per lactation. The upper limit for milk production per cow continues to increase. A challenge exists in developing nutrition programs for these herds. The goal is to attain efficient and profitable levels of milk production while maintaining herd health and reproductive performance. Evaluation of rations currently fed to high producing herds indicate that these rations are consistent with current nutrient requirement guidelines. Many high producing herds have average DMI > 4% of BW. Ration formulation principles and nutrient requirements used in development of feeding programs for high producing herds are similar to methods already in use. Optimizing DMI, optimizing rumen fermentation, and providing supplemental nutrients are key factors in meeting tissue nutrient demands in this formulation process. PMID- 8227649 TI - Milking the 30,000-pound herd. AB - The principles for milking the 13,600-kg (30,000-lb) cow are the same as for any other dairy cow: she should be milked gently, quickly, and completely with minimal machine stripping or over-milking. The application of these principles may differ, however, because high producing cows have 1) lower premilking stimulus requirements than low producers, 2) higher peak milking rates and higher average flow rates (yet longer times to milkout), 3) higher incidence of teat orifice lesions such as hyperkeratosis, and 4) higher risk of new mastitis infections. Existing national and international standards for construction and performance of milking systems may not be adequate to manage the higher expected flow rates through the milking unit and milklines. Such standards need to be reviewed and perhaps revised to ensure appropriate sizing and flow characteristics based on sound engineering principles and physiological requirements. Milking four times per day increases daily milk production compared with twice per day, and milking eight times per day increased daily production compared with four times per day. Cows milked more than four times per day might not require complete milking out at every milking. More frequent milking (or milking on demand) may be achieved with robot milkers, provided that robots can match the reliability of human milkers. PMID- 8227650 TI - The contribution of lactococcal starter proteinases to proteolysis in cheddar cheese. AB - The contribution of the lactococcal proteinase to proteolysis and flavor development in Cheddar cheese was investigated using the starter strains Lactococcus lactis ssp. lactis UC317, its proteinase-negative derivative FH041, and variants of UC317 modified in proteinase production, location, and specificity. Lactococcus lactis ssp. lactis FH041 was transformed by electroporation with plasmids pCI3601, pCI3602, or pNZ521. Plasmids pCI3601 and pCI3602 harbor the cloned proteinase genes of L. lactis ssp. lactis UC317 on a high copy number vector and, as such, encode an increased concentration of cell wall-associated and secreted enzymes, respectively. Plasmid pNZ521 contains the cloned proteinase genes from Lactococcus lactis ssp. cremoris SK11. Assessment of proteolysis and flavor development in Cheddar cheese made with these strains revealed that starter proteinases are required for the accumulation of small peptides and free amino acids in Cheddar cheese. Proteolysis was not enhanced by an approximately threefold increase in concentration of the lactococcal proteinase. The strain in which the proteinase remained attached to the cell wall appeared to contribute more to proteolysis than the strain that secreted the enzyme. Water-soluble peptides unique to Lactococcus lactis ssp. cremoris SK11 and L. lactis ssp. lactis UC317 were detected by PAGE and HPLC, respectively. Sensory evaluation showed that the flavors of all cheeses made with proteinase positive starters were similar, but cheeses made with proteinase-negative starters lacked flavor. PMID- 8227651 TI - Augmentation of macrophage phagocytic activity by cell-free extracts of selected lactic acid-producing bacteria. AB - Oral and intraperitoneal administration of lactic acid-producing bacteria can significantly augment the immune response in murine models; however, the immunopotentiating effects in these studies differ significantly. Murine macrophagelike cell line J774 was cultured in the presence of cell-free extracts of Lactobacillus acidophilus and Bifidobacterium longum, and the effect on macrophage function was evaluated by measurement of synthesis of selected enzymes and their ability to take up either acrylamide particles or live Salmonella typhimurium. Lysozyme activity of J774 cells was significantly decreased by cell free extracts of B. longum, but not of L. acidophilus, whereas extracts of both strains induced morphological changes and significantly enhanced phagocytosis of inert particles or viable Salmonella. Whole cell extracts of lactic acid producing bacteria are therefore capable of altering macrophage function in a strain-dependent manner. PMID- 8227652 TI - Effect of antioxidative lactic acid bacteria on rats fed a diet deficient in vitamin E. AB - Lactic acid bacteria, including Bifidobacterium, with antioxidative activity were selected by in vitro screening. The effect of the antioxidative activity was investigated by in vivo experiments using rats that were deficient in vitamin E. In the first stage of screening, 570 strains were examined; intracellular cell free extracts of 19 strains (16 lactobacilli, 2 streptococci, and 1 lactococci) had antioxidative activity as determined by an assay using rat liver microsomes and thiobarbituric acid. In the second stage of screening, 7 strains of lactobacilli showed over 70% inhibition of oxidation activity. The highest activity was obtained by heterofermentative Lactobacillus sp. SBT 2028. The effect of two strains, Lactobacillus sp. SBT 2028 and Lactobacillus casei ssp. rhamnosus SBT 2257, was evaluated for improvement of the condition of rats fed a diet deficient in vitamin E. Intracellular cell-free extracts of those two strains were also used for in vivo experiments. Hemolysis of red blood cells was inhibited in rats that were administered the extract of Lactobacillus sp. SBT 2028, which proved that the extract improved the vitamin E deficiency status. Antioxidative activity of an extract from L. casei ssp. rhamnosus SBT 2257 determined by hemolysis was relatively weak compared with the activity of Lactobacillus sp. SBT 2028 extract. PMID- 8227653 TI - Three-dimensional molecular modeling of bovine caseins: a refined, energy minimized kappa-casein structure. AB - A refined three-dimensional molecular model of kappa-casein has been produced using energy minimization techniques and a Kollman force field on a previously reported predicted three-dimensional structure. This initial model was constructed via molecular modeling techniques from sequence-based secondary structural prediction algorithms. Both the initial and refined structures agreed with global secondary structure analysis from vibration spectroscopy. The refined structure contained many of the features of the initial model, including two sets of antiparallel beta-sheet structures containing predominantly hydrophobic side chains, which could form interaction sites with alpha s1-casein. Two types of energy-minimized dimer and tetramer models are presented: 1) using Cys as potential intermolecular disulfide binding sites and 2) using the two sheets as possible hydrophobic self-association sites, without Cys interactions. All structures yielded good stabilization energies and are in agreement with chemical, biochemical, and physical chemical results obtained for kappa-casein. PMID- 8227654 TI - Effect of body condition score at calving on performance, some blood parameters, and milk fatty acid composition in dairy cows. AB - Effect of body condition score at calving and of change in body condition score on productive and reproductive performance, on lactation curve parameters, and on postpartum disease occurrence was investigated in 213 Italian Holstein-Friesian cows. Plasma FFA, glucose, and insulin concentrations and milk fatty acid composition were analyzed in 15 cows. The cows calving at highest body condition score lost more subcutaneous fat; condition score change did not exceed 1.05 units. Change in body condition score was positively associated with peak and total milk production. Occurrence of retained placenta was not related to the content of fat stores at calving. Glucose concentrations were relatively constant, FFA concentrations were highest, and insulin concentrations were lower in cows calving at the highest body condition score. Milk fatty acid composition reflected the different utilization of body fat stores. PMID- 8227655 TI - Postinsemination administration of receptal: follicular dynamics, duration of cycle, hormonal responses, and pregnancy rates. AB - In experiment 1, concentrations of LH, FSH, and progesterone, but not estradiol 17 beta, in blood serum were increased during 6 to 12 h after injection of 8 micrograms of receptal (GnRH agonist) administered on d 11 to 14 after estrus (d 0) and at first AI compared with saline treatment in lactating Holstein cows. Beginning 2 to 3 d after injection of receptal, concentrations of progesterone were increased for 3 d in nonpregnant cows and for 12 d in pregnant cows compared with controls of similar pregnancy status. Number of ovarian follicles determined by ultrasonography during 10 d after receptal was reduced, specifically those with antral diameters of > or = 10 mm. The dominant follicle in both groups began to decrease in diameter on the day following treatment, but the next dominant follicle began to increase in diameter 2.3 +/- .7 d later in receptal-treated cows than in controls, accounting for an increase in cycle duration of 2.5 +/- .8 d. In Experiment 2, a double-blinded study was conducted in eight herds in which cows (n = 1013) were AI at first detected estrus after 50 d postpartum and assigned randomly to receive either saline or 4, 8, or 12 micrograms of receptal on d 11 to 14 after first AI. Pregnancy rates were improved in one herd at all doses of receptal, but dose responses were inconsistent in remaining herds. A greater proportion of cows given receptal than controls returned to estrus after 24 d. Administration of a potent GnRH agonist altered number and distribution of ovarian follicles, increased cycle duration, and increased concentrations of progesterone without a consistent increase in pregnancy rates. PMID- 8227656 TI - Ovarian follicular waves in dairy cows after administration of gonadotropin releasing hormone at estrus. AB - A crossover design experiment was conducted to examine the effects of GnRH on hormonal concentrations and changes in ovarian structures of lactating dairy cows during the estrous cycle. Cows were assigned randomly to receive either 100 micrograms of GnRH or saline 12 h after estrus (d 0) was first observed. Ultrasonographic examinations of the ovaries performed daily throughout the estrous cycle detected either three (n = 7) or four (n = 6) follicular waves. Transient increases in FSH generally preceding the follicular wave containing the dominant (preovulatory) follicle, and the emergence of the dominant follicle before the subsequent estrus occurred later in GnRH-treated than in saline treated cows with either three or four follicular waves. Day of emergence of dominant follicles within follicular waves was preceded 83% of the time by a transient increase in FSH and was followed consistently by increased serum estradiol-17 beta, which generally reached peak concentrations concomitant with the peak diameter of the dominant follicle. Although clearly defined pulses were detected in 19 out of 26 sampling periods in d 7 and 14, number of FSH pulses and percentage of cows with pulses of FSH were similar in GnRH-treated and saline treated cows with three or four follicular waves. In conclusion, treatment of cows with GnRH 12 h after the onset of estrus delayed the appearance of the next ovulatory follicle in cows with either three or four follicular waves. Regardless of treatment, emergence of dominant follicles was preceded in most cases by a transient increase in FSH and followed consistently by increased estradiol-17 beta. PMID- 8227657 TI - Short- and long-term production losses and repeatability of clinical mastitis in dairy cattle. AB - Between 1985 and 1990, a study of 5313 lactations of 2477 Black and White cows was carried out. A stepwise least squares method was used to obtain unbiased estimates of milk, fat, and protein losses that were due to clinical mastitis and the carry-over effect from the previous lactation. Logistic regression was used to estimate the probability that a cow would have clinical mastitis in the next month. The effect of clinical mastitis on production within one lactation was estimated at 527 kg of milk (8.1%), 22.7 kg of fat (8.0%), and 13.7 kg of protein (6.2%) for > or = 3 cases of clinical quarters in the second lactation. One or 2 cases of clinical quarters in a lactation did not significantly affect the production in the next lactation. The negative carry-over effect of > or = 3 cases of clinical quarters was estimated at 381 kg of milk (5.9%), 23.7 kg of fat (8.4%), and 10.1 kg of protein (4.6%) up to and including mo 8 of the second lactation. The fat content in milk produced after the onset of mastitis decreased, and the protein content increased. The risk of clinical mastitis infection in the following month was influenced by month of lactation (a higher risk early in lactation), lactation number (risk increased with lactation number), production level (higher risk for high producing cows), number of clinical quarters in the previous lactation, number of clinical quarters in the previous months of the current lactation, and occurrence of clinical mastitis in the current month. PMID- 8227658 TI - Bovine acute mastitis: effects of intravenous sodium salicylate on endotoxin induced intramammary inflammation. AB - Effects of the nonsteroidal antiinflammatory agent sodium salicylate on endotoxin induced mastitis were evaluated in lactating cows. Escherichia coli endotoxin was administered to a mammary quarter 1 h after initiation of a 12-h i.v. infusion of sodium salicylate. Milk SCC, BSA concentrations in milk, mammary inflammation, rectal temperature, appetite, milk production, and plasma and lymph PGF2 alpha were monitored. Gross mammary inflammation was not reduced by salicylate infusion, nor did sodium salicylate prevent increased milk SCC or BSA concentrations in milk, although treatment tended to decrease the magnitude of these responses. Sodium salicylate decreased subcutaneous abdominal vein PGF2 alpha metabolite, and PGF2 alpha metabolite tended to be reduced in lymph during the acute phase of inflammation. The increased rectal temperature after endotoxin infusion was reduced in cows treated with sodium salicylate. Appetite was reduced after endotoxin infusion in untreated cows and those treated with sodium salicylate. Milk production declined after endotoxin challenge in all cows. Although sodium salicylate did not substantially reduce mammary inflammation, it had an antipyretic effect and reduced PGF2 alpha metabolite in mammary blood. PMID- 8227659 TI - Relations between on-line electrical conductivity and daily milk production on a low somatic cell count farm. AB - To study the relation between on-line electrical conductivity and daily milk production, data of 1389 cow days were analyzed. After correction for cow effects and DIM, a rise of 1 mS of the mean electrical conductivity caused a decline of .88 kg/d in milk production. A rise of 1 ln(SCC) unit was associated with an additional decline of .54 kg/d in milk production. In cows without clinical mastitis during the test period, the losses associated with mean electrical conductivity and ln(SCC) were 1.06 and .45 kg/d of milk production, respectively. Electrical conductivity and SCC were associated with daily production loss; the effects were additive. Therefore, electrical conductivity and SCC can be utilized as indirect tests of subclinical mastitis. PMID- 8227660 TI - Forms of lactoferrin: their antibacterial effect on enterotoxigenic Escherichia coli. AB - The antibacterial effects of various forms of lactoferrin on enterotoxigenic strains of Escherichia coli were tested in vitro using a microassay for bacterial growth. Native and apo-lactoferrin exhibited variable activity against 19 strains, whereas holo-lactoferrin had no effect. At a concentration of .2 mg/ml of apo-lactoferrin, strains could be distinguished as either sensitive or resistant to inhibition. Zinc-saturated lactoferrin was as inhibitory as apo lactoferrin when sensitive and resistant strains were tested over the concentration range .04 to 1.0 mg/ml of lactoferrin. A bactericidal effect was observed for native, apo-, and Zn-saturated lactoferrin against some sensitive strains. The antibacterial activity of apo-lactoferrin depended on bacterial inoculum size and was not enhanced by the addition of lysozyme. Addition of holo lactoferrin or cytochrome c diminished the antibacterial effect of apo lactoferrin, whereas addition of BSA had no effect. Resistance to inhibition by lactoferrin was not related to the production of bacterial siderophores. PMID- 8227661 TI - Circadian and ultradian rhythms of peripheral cortisol concentrations in lactating dairy cows. AB - To investigate possible circadian and ultradian periodicities for plasma cortisol in lactating dairy cows, integrated 15-min blood samples taken sequentially over 48 h from six cows were analyzed by radioimmunoassay. The cows were housed in an environmental chamber at about 19 degrees C, 50% relative humidity, and 16 h of light and 8 h of darkness (lights on at 0700 h); fed daily at 0900 h; and milked at 0830 and 2000 h. Peripheral concentrations of cortisol for all six cows exhibited weak circadian rhythms--average minima were 3.1 ng/ml at 1800 h, maxima were 4.5 ng/ml at 0530 h--and strong ultradian rhythms with periods around 120 min. Peak to trough amplitudes of ultradian rhythms, 1 to 17 ng/ml, varied among and within cows over time. No direct relation between ultradian peaks and milking or feeding was apparent. Spectral analysis indicated that most power (variation) was centered around 1 cycle every 2 h, i.e., around a period of 120 min. This very strong, easily observable, ultradian rhythm indicates that an oscillation with a period around 120 min is probably intrinsic to mechanisms regulating peripheral glucocorticoid concentrations. PMID- 8227662 TI - Blood polymorphonuclear leukocyte chemotaxis during experimental Escherichia coli bovine mastitis. AB - The relationship between the severity of experimental Escherichia coli mastitis and the chemotactic response of blood polymorphonuclear leukocytes was investigated before and during mastitis. Experimental E. coli mastitis was induced in 10 healthy cows by inoculation of the rear right quarters with 10(3) cfu of E. coli. Cows were classified into two groups based on the severity of the mastitis. Bacterial growth in the inoculated quarter was used as parameter that indicated severity. Before and during experimental mastitis, the chemotactic response and the number of circulating polymorphonuclear leukocytes were greater for the moderately diseased cows than for the severely diseased cows. During the first 24 h of the experimental mastitis, the chemotactic response of polymorphonuclear leukocytes decreased in both groups. Recovery of the chemotactic response of white blood cells was more rapid in moderately diseased cows than in severely diseased cows. Possibly, the larger proportion of band neutrophils (the less chemotactically active band neutrophils) partially accounts for the lower chemotactic response of the circulating polymorphonuclear leukocytes during experimental mastitis in the severely diseased cows. PMID- 8227663 TI - Induction of estrus after thyroidectomy in nonlactating Holstein cows. AB - Hypothyroidism reportedly decreased sexual behavior in several species. To confirm the role of hypothyroidism in the expression of estrus in cattle, our objective was to determine whether estrus could be induced in ovariectomized cows without intact thyroid glands using estradiol benzoate and progesterone. Eight nonlactating, nonpregnant Holstein cows were thyroidectomized and ovariectomized, and another four cows were ovariectomized only. All cows were given either estradiol benzoate in a crossover design with two replicates during 4 consecutive wk. Cows were observed continuously for estrus from 9 to 45 h after treatment with gonadal steroids. Intensity of estrus was assessed by quantitative measurement of the frequencies of mounting activity and standing behavior. The percentage of cows showing estrus was influenced by thyroidectomy-ovariectomy (78.1%) compared with ovariectomy (31.1%) but was unaffected by hormonal treatment. In contrast, other measures of estrous behavior (duration of standing estrus and frequencies of mounting and standing activities) were unaffected by thyroid status or by hormonal treatments. We concluded that ovariectomized cows without intact thyroid glands exhibit estrus and that the exhibited behavior is similar in intensity and duration to that in ovariectomized cows treated with gonadal steroids. Hypothyroidism may not cause failure to express estrus. PMID- 8227664 TI - Observations on in situ degradation of forage cell components in alfalfa and Italian ryegrass. AB - The rate and extent of degradation of forage feed fractions contained in alfalfa and Italian ryegrass hays were determined. Nylon bags filled with 4 g of each forage were suspended in the rumen of two cannulated cows immediately before feeding and incubated for 10 different times (0, 2, 4, 8, 12, 24, 48, 72, 120, and 168 h). The alfalfa hay, which had lower NDF, showed a lower extent, but a higher rate, of NDF degradation than the Italian ryegrass (41.1 vs. 59.8% and 4.64 vs. 2.91%/h, respectively). Alfalfa cell walls were degraded more rapidly than Italian ryegrass even though their lignin content was higher. The hemicellulose fermentation of alfalfa showed a longer lag time (13 h) and an undegradable fraction nearly twice that for Italian ryegrass (63.3 vs. 37.1%). Cellulose from alfalfa was degraded at a higher rate than NDF or ADF, indicating that cellulose may be the primary site of hydrolysis of the cell wall in the rumen. Calculations based on in situ degradability indicate that alfalfa can have a higher inclusion than Italian ryegrass in diets for dairy cows because of lower NDF and greater availability of cell contents. PMID- 8227665 TI - In situ dry matter, protein, and fiber degradation of perennial forages. AB - Eight forages (alfalfa, birdsfoot trefoil, red clover, bromegrass, orchardgrass, perennial ryegrass, quackgrass, and timothy) at three maturities were evaluated for ruminal DM, CP, and NDF degradation kinetics. Duplicate dacron bags were incubated for 0, 3, 6, 10, 13, 25, 48, and 72 h in two late lactation Holstein cows fitted with ruminal cannulas over eight experimental periods. Species and maturity effects were observed for soluble, slow, and undegraded fractions; degradation rate; and ruminally degradable DM, CP, and NDF. Significant species by maturity interactions also were evident for fractions and ruminal degradabilities of DM, CP, and NDF. Legumes exhibited more extensive ruminal DM degradation than did grasses. No clear trends were evident in rate of ruminal CP degradation between legumes and grasses. Mature grasses were lowest in ruminally degradable CP. Legumes exhibited a higher undegraded fraction and faster degradation rate of slowly degraded NDF fractions, resulting in similar ruminally degraded NDF for legumes and grasses. Extensive differences in ruminal degradation kinetics existed between perennial legume and grass species and maturities. Species by maturity interactions were diverse, making categorization of degradation characteristics of legumes and grasses difficult. PMID- 8227666 TI - Effectiveness of neutral detergent fiber in whole cottonseed and dried distillers grains compared with alfalfa haylage. AB - Sixteen Holstein cows in midlactation were used in a 4 x 4 Latin square design to determine the effect of replacing alfalfa NDF with NDF from whole, linted cottonseed or dried distillers grains. Low and high fiber control diets (13 and 19% of dietary DM from alfalfa haylage NDF, respectively) were compared with diets designed to contain 13% of DM from alfalfa haylage NDF plus 6% of DM from either cottonseed NDF or distillers grains NDF. Dry matter intake, milk yield, and milk protein yield were lower from the high fiber control diet. Milk fat percentage was lower for the low fiber control diet. The cottonseed diet was equal to the high fiber control diet in stimulating rumination. Rumen acetate to propionate ratio was higher for the high fiber control and cottonseed diets. Replacing alfalfa with either of these high fiber by-product feeds increased feed intake and yields of milk fat and protein. The effectiveness of the NDF in distillers grains and cottonseed was not significantly different from that of alfalfa NDF for maintaining milk fat yield. Whole cottonseed and dried distillers grains appear to be good sources of effective fiber for maintaining milk fat test when they are substituted for alfalfa haylage fiber in lactating cow rations. PMID- 8227667 TI - Quality evaluation and concentrate supplementation of rotational pasture grazed by lactating cows. AB - The effects of pasture quality and the amount and nutrient composition of a concentrate supplement on performance of lactating cows were investigated under a controlled rotational grazing system. Pasture samples were collected for two consecutive summers, and the relationship was positive between wet chemistry and near infrared reflectance spectroscopy for CP, ADF, and NDF. During yr 2, 54 Holstein cows averaging 82 d postpartum were used in a randomized block design to evaluate three methods of supplementing grain for a 24-wk trial. A 16.2-ha area of primarily orchardgrass pasture was divided into paddocks of approximately .40 ha each. The three supplemental grain rations were: 1) 1 kg of grain/3 kg of milk; 2) 1 kg of grain/4 to 5 kg of milk adjusted weekly, depending upon the quantity of pasture available; and 3) 1 kg of grain/4 to 5 kg of milk. The amount fed in ration 3 was adjusted weekly, depending upon the quantity of pasture available, and reformulated biweekly based on analysis of pasture. No differences existed among treatments for milk production (24.7, 23.2, and 24.1 kg/d) or 3.5% FCM (25.3, 24.8, and 24.9 kg/d) over the 24-wk trial. Cows fed more grain (ration 1) gained more BW. Adjusting nutrient density of the grain to changes in pasture quality resulted in similar milk production and a lower ration cost. PMID- 8227668 TI - Effects of inert fat on energy balance, plasma concentrations of hormones, and reproduction in dairy cows. AB - The objective was to determine the effects of dietary inert fat on estimated energy balance, hormones in plasma, and reproduction during early lactation. From wk 0 to 12 postpartum, 14 pluriparous Holstein cows were fed individually a TMR, and blood samples were taken twice weekly for quantification of IGF-I, progesterone, and cholesterol. During wk 5 to 12, one-half of the cows remained on the TMR, and the other half were fed the TMR containing inert fat at 1.8% of dietary DM. Estrous behavior was monitored twice daily, and body condition scores were recorded every 4 wk. Cows fed inert fat between wk 5 and 12 postpartum had similar concentrations of IGF-I in plasma but greater luteal phase progesterone secretions than cows fed the control diet. Total cholesterol in plasma also was greater in cows fed inert fat than in cows fed the control diet. Intervals to first, second, and third ovulation or estrus did not differ among cows fed control or inert fat diets. Body condition scores, daily DMI, and milk production were not affected by dietary inert fat. Inert fat fed to cows between wk 5 and 12 postpartum did not affect ovulatory activity but may enhance luteal activity. PMID- 8227669 TI - Rumen fermentation and lactation performance of cows fed roasted soybeans and tallow. AB - Sixteen multiparous Holstein cows were in a replicated 4 x 4 Latin square design with 21-d periods to examine the effects of incremental tallow addition to diets containing whole roasted soybeans on rumen fermentation and lactational performance. Total mixed rations were fed for ad libitum intake and contained, on a DM basis, 33% alfalfa silage, 12% corn silage, 14% roasted soybeans, and 41% concentrate based on ground corn and soybean meal. Treatments were 0, 1, 2, or 3% supplemental tallow. Diets contained 20% CP and ranged from 1.68 to 1.82 Mcal NEL/kg of DM. The DMI, milk yield, milk protein and fat yields, milk fat percentage, rumen acetate: propionate ratio, and in situ forage DM disappearance did not differ among treatments. A small linear decrease occurred in milk protein percentage as tallow feeding was increased (2.89 to 2.86%). Tallow supplementation increased total VFA concentration in rumen fluid and resulted in a linear decrease in rumen pH (6.17 to 5.99). Supplementation of 1 to 3% tallow to diets containing 2.8% supplemental fat from whole roasted soybeans had minimal negative effects on rumen fermentation and did not influence lactational performance. PMID- 8227670 TI - Lupins (Lupinus albus) as a protein supplement for lactating Holstein dairy cows. AB - Three experiments were conducted to determine optimal substitution of lupin for soybean meal in dairy cattle diets; the effects of whole versus ground lupins and the effects of whole lupins versus whole soybeans on milk production were examined. In the first experiment, 57 cows were assigned randomly to treatments in blocks of five by calving order within parity. Treatments began 22 d postpartum and continued through d 140. Diets were isonitrogenously balanced using soybean meal as control: lupins replaced 0, 25, 50, 75, and 100% of supplemental protein. Cows that consumed 75% of supplemental protein as lupins produced more 3.5% FCM and fat than cows that consumed the control diet. In a second experiment, 30 primiparous cows were assigned randomly by calving order to either ground or whole lupins (60% of supplemental protein). Treatments began 32 +/- 3 d after calving and lasted for 84 d. Cows that consumed ground lupins produced more milk and 3.5% FCM than cows that consumed whole lupins. In the third experiment of 84 d, 28 primiparous midlactation cows were assigned randomly to diets containing whole lupins or whole soybeans to supply 1 kg of supplemental protein. Cows fed whole lupins produced milk with higher fat content, more 3.5% FCM, and more fat than cows consuming whole soybeans. Lupins are an acceptable supplemental protein source for dairy cows. PMID- 8227671 TI - Ryegrass versus corn starch or beet pulp fiber diet effects on digestion and intestinal amino acids in dairy cows. AB - Changes in digestion and AA supply in dairy cows were studied when fresh grass was partly replaced by concentrate mixtures based either on corn starch or sugar beet pulp fiber. Treatments were tested in a Latin square utilizing three lactating cows' with ruminal and intestinal cannulas. Partial replacement of grass decreased CP digestibility. When high starch concentrate was fed, NDF digestibility was lower than that of the high fiber diet, mainly because of decreased ruminal digestion of NDF. For the high starch concentrate, 39% of the ingested starch escaped ruminal fermentation. Although less OM was fermented in the forestomachs on high starch concentrate, the duodenal AA N flow was higher than for the high fiber concentrate. The proportion of microbial protein was unaffected; thus, efficiency of microbial synthesis was estimated to be higher when high starch concentrate was fed. PMID- 8227672 TI - Effect of source and amount of protein on milk production in dairy cows. AB - Forty multiparous Alpine does (mean BW of 61.5 kg) were utilized in a 13-wk trial to investigate the effects of a TMR differing in CP amount (13 or 17%) and source (solvent-extracted soybean meal or heat-treated soybean meal with or without urea) on lactational performance. Protein supplements contributed 30% of the N in 13% CP diets and 50% of the N in 17% CP diets. All diets were isoenergetic (2.5 Mcal of metabolizable energy/kg of DM) and were fed for ad libitum intake for the entire trial. Mean DMI (2.88 kg/d), milk production (2.65 kg/d), milk fat (4.05%), milk protein (2.68%), milk lactose (4.54%), and milk SNF (7.81%) did not differ among dietary treatments. Plasma urea N was greater (23.2 vs. 10.9 mg/dl) in does receiving the 17% CP diets; however, blood hematocrit (27.4%), beta hydroxybutyrate (843 microM), plasma glucose (68.8 mg/dl), NEFA (600 mu eq/dl), and plasma total protein (74.% g/L) were not significantly affected by treatment. The apparent absence of a dietary effect on lactational performance may be due to the high DMI of the does (4.7% when expressed as DMI per kilogram of BW) and high CP intake providing a surfeit of protein relative to requirements. PMID- 8227673 TI - Copper status and requirement during the dry period and early lactation in multiparous Holstein cows. AB - Eighteen multiparous Holstein cows were assigned randomly to three treatments at the beginning of the dry period (8 wk before expected parturition). Treatments were: 1) the basal diet containing 5.5 ppm of Cu (control), 2) the basal diet supplemented with 10 ppm of Cu, and 3) the basal diet supplemented with 20 ppm of Cu. The objectives were to measure the changes of Cu and other trace mineral element concentrations in blood and liver from the onset of the dry period (approximately 8 wk prepartum) to 8 wk postpartum and to assess the requirement of Cu during this time. Liver Cu concentration in the control group declined continuously during the 8-wk dry period, and the nadir occurred at parturition. This decline was prevented by dietary Cu supplementation of 10 or 20 ppm. Liver Cu concentration in the control group started to increase slowly after the dramatic decline. Liver Zn concentration changed cubically as a function of week during the treatment period. Plasma Cu and Zn exhibited a quadratic pattern as a function of week. The plasma Cu concentration was lowest 5 wk prior to parturition. PMID- 8227674 TI - Effects of yeast on dairy cow performance, ruminal fermentation, blood components, and milk manufacturing properties. AB - Effect of dietary yeast culture on milk production and composition, ruminal fermentation, blood parameters, and milk manufacturing properties of 24 midlactation Holstein-Friesian cows was determined in a 6-wk experiment. The control diet (DM) consisted of 30% corn silage, 22% alfalfa hay, and 48% concentrate. After a 2-wk preliminary period, cows were assigned in equal numbers to either 0 or 10 g/d of yeast culture for the remainder of the 4-wk study on the basis of age, DIM, and pretrial milk production. Production of milk (26.2 vs. 25.4 kg/d), FCM (23.6 vs. 21.6 kg/d), and milk fat (.90 vs. .78 kg/d) was increased significantly by dietary yeast culture. No differences were significant for milk composition. Molar proportion of acetate and acetate:propionate ratio in ruminal liquor tended to be higher in cows fed yeast culture. Total VFA concentration in ruminal fluid was not different between treatments. Manufacturing properties of milk and blood plasma components were not affected adversely by added dietary yeast culture. PMID- 8227675 TI - Lactation potential as a criterion for strategy of feeding total mixed rations to dairy cows. AB - A trial was conducted from freshening to 30 wk postpartum to evaluate assignment of cows into production potential groups based on early lactation performance to optimize the time for switching from high to medium nutrient concentration TMR. Cows received a TMR formulated for early lactation until wk 4 of lactation and a high nutrient concentration TMR until 13, 19, or 25; the high TMR was followed by a medium nutrient concentration TMR until the end of the trial. Cows were assigned to high, medium, or low potential groups separately for parities 1, 2, and > or = 3 based on daily 3.5% FCM production as a percentage of BW measured during wk 6 and 7 of lactation. Low potential cows tended to partition energy intake into BW gain earlier in lactation than medium and high potential cows. For low potential cows, an early transfer to medium TMR moderated BW gain with no negative effect on production. High potential cows maintained their BW on the high ration; however, early transfer negatively affected production. Classification of individual cows into potential groups based on early lactation performance can serve as a useful tool for TMR feeding strategy later in lactation. PMID- 8227676 TI - Effect of insulin-induced hypoglycemia on cessation of macromolecular transport in the neonatal calf. AB - The effect of hypoglycemia on the absorption of colostral Ig was studied in 12 neonatal calves. Hypoglycemic calves received 1 cc (100 units) of insulin at birth; control calves received 1 cc of saline. Two dietary regimens were imposed; colostrum was fed at birth or at 24 h. Glucose, insulin, and IgG were measured in venous blood. Insulin induced decreases in circulating glucose in insulin-treated calves from 12 to 42 h postnatally; insulin concentrations were significantly higher from 12 through 24 h. Concentrations of glucose also were decreased in calves deprived of initial colostrum compared with those fed at birth over the same period, although insulin concentrations were not different. Time of closure was delayed in insulin-treated calves relative to saline-treated calves (40 and 29 h, respectively) and in calves deprived of initial colostrum relative to those fed at birth (45 and 23 h, respectively). Glucose availability may influence timing of closure in the calf, although the decreased rate of absorption of IgG in hypoglycemic calves prevents them from realizing any benefit in peak IgG concentrations higher than those of their normoglycemic counterparts. PMID- 8227677 TI - Postweaning age effects on rumen fermentation end-products and digesta kinetics in calves weaned at 5 weeks of age. AB - Four rumen-cannulated calves were used in a block design to study changes in rumen fermentation end-products and rumen digesta kinetics after weaning. Calves were weaned at 5 wk of age and had access to dry feed from d 3 of age. Dietary DM consisted of 85% concentrate and 15% forage. The experiment was divided into three periods: 2, 4, and 8 wk after weaning. Rumen fluid samples collected each period were analyzed for pH, VFA, NH3 N, bacterial DM concentrations, and protozoa numbers. Calves were dosed ruminally with Co-EDTA and Yb-mordanted grain and then sampled for estimation of rumen fluid and particle turnover rates during each period. Average daily gain, DMI, and BW gain increased over the three periods by 106, 81, and 42%, respectively. Rumen pH increased .32 units, and rumen volume increased twofold, during the three periods. Rumen NH3 N concentration and diurnal NH3 N variation changed significantly with age. Bacterial DM concentrations, VFA, and digesta kinetics in the rumen did not change with age. We conclude that the metabolic and physical changes that take place in the rumen of young calves after weaning are evidence of gradual development and yield important information for further study on degradation and metabolism of dietary components in the rumen. PMID- 8227678 TI - Effect of postweaning age on rate of in situ protein disappearance in calves weaned at 5 weeks of age. AB - Four rumen-cannulated calves were used in a block design to evaluate the in situ rate of protein disappearance in the rumen of soybean meal, heat-treated soybean meal, corn gluten feed, and corn distillers grains at 2, 4, and 8 wk after weaning. The rates of degradation of the four feeds also were evaluated in an in situ experiment using lactating cows and in an enzymatic in vitro experiment using Streptomyces griseus to determine whether the in vitro method could predict protein degradation in the rumen of calves. An interaction between protein source and age after weaning occurred for rate of CP disappearance. For soybean meal and corn gluten feed, the rate of disappearance of CP increased with age, but no age effect on rate of CP disappearance occurred for heat-treated soybean meal or distillers grains. Rates of CP disappearance between cows and calves were similar for distillers grains and corn gluten feed, but no similarities existed between the in vitro and in situ methods. During the first 4 wk after weaning, feedstuffs high in degradable protein have lower rumen degradation rates than they do at 8 wk after weaning. Rumen degradation rates of low degradable proteins do not change during the first 8 wk after weaning. In addition, the cow in situ and in vitro methods were not suitable for prediction of rumen protein degradability of early weaned calves. PMID- 8227679 TI - Genetic evaluation of length of productive life including predicted longevity of live cows. AB - Complete longevity data are available too late for most sire selection. Earlier selection is possible using correlated traits, nonlinear evaluation of censored data, or predicted longevities for live cows in addition to completed longevity data. Completed longevity was defined as total months in milk by 84 mo of age. Predicted longevity was computed by multiple regression from cows alive at six different ages. Variables included age at first calving, standardized first lactation milk yield (optional), lactation status (dry or milking), current months in milk, current months dry, and cumulative months in milk. Completed longevity data for dead cows were then merged with predicted longevity data for live cows. A total of 1,984,038 Holstein cows born from 1979 to 1983 were included and represented 1911 sires, each with at least 70 daughters. Heritability of longevity increased gradually from .03 at 36 mo to .08 at 84 mo. Phenotypic correlations of early with completed longevity ranged from .59 to .99; genetic correlations ranged from .92 to 1.00. Inclusion of yield for projection increased heritability at 36 mo substantially but decreased genetic correlation with complete longevity information. Expansion and weighting factors will allow predicted records to be used in longevity evaluations, which is similar to the procedures that allow predicted 305-d yields to be included in yield evaluations. PMID- 8227680 TI - Nonlinear mixed model analyses of five production disorders of dairy cattle. AB - A standardized threshold model was used to estimate herd variability and the impact of some risk factors on the incidence of retained placenta, metritis, ketosis, ovulatory dysfunction, and mastitis. Data consisted of records for 18,110 Swedish Red and White cows in 924 herds and 14,940 Swedish Friesian cows in 772 herds. Only first, second, and third party cows were included, and the breeds were analyzed separately. Herds were considered to be random effects, and variance components were estimated by the counterpart of REML for a threshold model. Estimated intraherd correlations varied from .04 to .18 and were lowest for the incidence of retained placenta (for both breeds) and highest for ketosis and ovulatory dysfunction for Swedish Red and White and Swedish Friesian breeds, respectively. Parity increased predicted lactational incidence risks for all disorders except metritis, but season of calving had no profound effect. Dystocia and stillbirth increased the risk of retained placenta; dystocia and retained placenta increased the risk of metritis. The risk of ketosis was not influence by any of the considered risk factor diseases, and the risk of mastitis was influenced only by retained placenta for Swedish Red and White cows. For both breeds, the risk of ovulatory dysfunction was increased by metritis and ketosis. PMID- 8227681 TI - Efficacy evaluations on five chlorhexidine teat dip formulations. AB - Three developmental postmilking teat dip formulations containing chlorhexidine digluconate were evaluated against Staphylococcus aureus and Streptococcus agalactiae in sequential experimental exposure trials. Two additional commercial chlorhexidine digluconate teat dip products were evaluated in natural exposure trials. Under conditions of experimental challenge, the developmental formulations were efficacious against Staph. aureus but did not significantly reduce incidence of new IMI by Strep. agalactiae. None of the three formulations of a conventional germicide used as teat sanitizers effectively reduced incidence of new Strep. agalactiae IMI under experimental challenge conditions. In the natural exposure trials with negative controls, a .35% chlorhexidine teat sanitizer had efficacy of 88.7% against Staph. aureus and 51.4% against Strep. agalactiae. The .5% chlorhexidine product reduced Staph. aureus and Strep. agalactiae IMI by 86 and 56%, respectively. PMID- 8227682 TI - Physiological role of antioxidants in the immune system. AB - Diets contain naturally occurring antioxidant compounds that can stabilize highly reactive, potentially harmful molecules called free radicals. Free radicals are generated during normal cellular metabolism and result from the metabolism of certain drugs or xenobiotics. Exposure to UV light, cigarette smoke, and other environmental pollutants also increases the body's free radical burden. The harmful activities of free radicals are associated with damage to membranes, enzymes, and DNA. The ability of antioxidants to destroy free radicals protects the structural integrity of cells and tissues. This review focuses on data indicating that the functions of the human immune system depend on the intake of micronutrients, which can act as antioxidants. Recent clinical trials have found that antioxidant supplementation can significantly improve certain immune responses. Specifically, supplementation with vitamins C, E, and A or beta carotene increased the activation of cells involved in tumor immunity in the elderly. Supplementation with the antioxidant vitamins also protected immune responses in individuals exposed to certain environmental sources of free radicals. Supplementation with vitamin A, a relatively weak antioxidant, decreases morbidity and mortality associated with measles infections in children. PMID- 8227683 TI - Role of vitamin E and selenium in host defense against mastitis. AB - Vitamin E and Se are essential nutrients that share common biological activities. Deficiencies in either of these micronutrients have been related in increased incidence and severity of mastitis. A known physiological consequence of alpha tocopherol or Se deficiency is reduced neutrophil activity. Vitamin E and the Se containing enzyme, glutathione peroxidase, and antioxidants that protect neutrophils from the destructive action of toxic oxygen molecules necessary for intracellular kill of ingested pathogens. Dietary supplementation of cattle with Se results in a more rapid neutrophil influx into milk following intramammary bacterial challenge and increased intracellular kill of ingested bacteria by neutrophils. Dietary supplementation of early lactation cows with vitamin E results in increased bactericidal activity by bovine blood neutrophils. Recently completed trials have shown that subcutaneous injections of vitamin E approximately 10 and 5 d prior to calving successfully elevated neutrophil alpha tocopherol concentrations during the periparturient period and negated the suppressed intracellular kill of bacteria by neutrophils that is commonly observed at calving. PMID- 8227684 TI - Role of carotenoids in the immune response. AB - The well-established provitamin A activities of certain carotenoids have hampered past research into their possible specific functions. More recently, research interests on carotenoids have been revived, largely because of their anticancer activities. beta-Carotene and other carotenoids have been reported to possess immunomodulatory activities in humans and animals. These carotenoids enhance lymphocyte blastogenesis, increase the population of specific lymphocyte subsets, increase lymphocyte cytotoxic activity, and stimulate the production of various cytokines. In addition, carotenoids also stimulate the phagocytic and bacteria killing ability of blood neutrophils and peritoneal macrophages. The action of these carotenoids is widely accepted to be independent of their provitamin A activity. The immunostimulatory action of carotenoids may be translated into improved health, including mammary and reproductive health in dairy cattle. More studies are needed to establish fully the beneficial effects of supplementation of different carotenoids on the health of dairy cattle. Furthermore, studies on carotenoids other than beta-carotene are needed. PMID- 8227685 TI - Oxidative stress, antioxidants, and animal function. AB - Reactive oxygen metabolites generated during normal metabolism and metabolism stimulated by xenobiotics can enter into reactions that, when uncontrolled, can impair performance of dairy cows. Direct effects include peroxidative changes in membranes and other cellular components. Indirectly, competitive consumption of reducing equivalents can interfere with important metabolic functions and divert glucose from other pathways by inducing the monophosphate shunt. Normally, the body is protected by a wide range of antioxidant systems working in concert. Metal catalysts of oxidative reactions are removed in intracellular fluids by metal-binding macromolecules. Superoxide dismutases, glutathione peroxidase, and catalase within cells remove superoxide and peroxides before they react with metal catalysis to form more reactive species. Finally, peroxidative chain reactions initiated by reactive species that escaped enzymatic degradation are terminated by chain-breaking antioxidants, including water-soluble ascorbate, glutathione, and urate and lipid-soluble vitamin E, ubiquinone, and beta carotene. To optimize performance, oxidative stress in high producing cows must be controlled by supplying all known antioxidant nutrients and by minimizing effects of substances that stimulate reactive oxygen metabolites. PMID- 8227686 TI - Pins for direct restorations. AB - Self-threading dentine pins permit the retention of large complex direct restorations but there are problems associated with their placement. Strain and crazing of dentine following pin insertion and pulpal and lateral perforations are common. Perforations can be avoided by operator awareness of tooth morphology. Strain and crazing has been found to be minimized by unscrewing the pin slightly after insertion, by using pins with a tap thread, and by using the smallest pin possible. Twist drill form and dulling affects the pin channel shape which in turn influences pin seating. A lack of standardization of pin and twist drill diameter and length has been implicated as the cause of poor pin retention. Manufacturers, in an attempt to standardize the depth of penetration of pins, have incorporated shoulders at the midpoint of the pin, which has met with varying success. More research in the area of limiting pin penetration is necessary, as well as attempts to improve the quality control of pin and twist drill manufacture. PMID- 8227687 TI - Marginal adaptation. AB - A critical review of studies assessing the marginal adaptation of direct placement, plastic restorations is presented. The effects upon adaptation of cavity design and location of cavity margins are examined, together with the effects of differing placement and finishing techniques. Both the choice of restorative material and the use of liners/bases are shown to influence the quality of restoration margins. Techniques used in both in vivo and in vitro assessment are reviewed, however it appears that a wide variety of methodologies exist and the establishment of standard, published criteria for the qualitative assessment of marginal adaptation is recommended. PMID- 8227688 TI - Bond strength of denture teeth to acrylic bases. AB - The literature relating to the determination of the bond strength of plastic denture teeth to acrylic resin denture material is reviewed. The papers are presented in chronological order with information on specimen preparation, batch sizes and methods of testing. The lack of uniformity in experimental techniques and the diverse range of products assessed makes recommendations for laboratory practice difficult to formulate. One consistent observation is that tooth surface contamination with wax decreases the bond strength between the teeth and the denture base material. A universal testing method needs to be formulated to replace the various techniques now employed. PMID- 8227689 TI - An analysis of durability data on post and core restorations. AB - An attempt was made to review the dental literature since 1970, presenting clinical data of posts and core restorations, by the use of a meta-analytic procedure. Fourteen publications were found that contained durability data on post and core systems. According to the exclusion criteria 11 studies were excluded for further analysis of the data. The survivals of these studies varied from 98.6% survival after a follow-up period of more than 10 years (F.S. Weine et al. 1991, J. Endodont. 17, 293-297) to 77.6% survival after a mean follow-up period of 5.2 years (D.H. Roberts, 1970, Br. Dent. J. 128, 117-124). The data of the three selected studies were reconstructed according to the method of Kaplan Meier but could not be combined for an overall survival assessment because the study characteristics were too heterogeneous. Therefore the survivals were presented separately. The reconstructed survivals of the studies after 6 years were 81% survival (standard error 6%) for resin composite build-ups in combination with screw posts and 91% survival for two studies including cast posts and cores (standard errors respectively 3% and 2%). Although it was not meaningful to combine the data for an overall survival, the meta-analytic method of this study revealed homogenic survival data and as such the followed procedure is considered to be an appropriate method to obtain insights in clinical durability data. PMID- 8227690 TI - Ultrasonic removal of dentine pins. AB - Replacement of pin-retained amalgam restorations is complicated by the loss of prime pin sites and remaining pin sites posing anatomical problems. Self threading dentine pins can be removed with an ultrasonic handpiece, and the pin site reused with a large pin. Retention of replacement pins is similar to original pins. Pin removal after experimental placement takes a considerable time (< 15 min), but pin removal from extracted pin-restored teeth is much quicker (mean of 45 s). The reuse of primary pin sites allows uncomplicated replacement of pin-retained amalgams, and is a useful clinical technique. PMID- 8227691 TI - A computer-aided study of speaking spaces. AB - A computer-aided system was devised to investigate the speaking space. Thirty native Cantonese speakers with Class I occlusion were selected and test sentences were designed for speech analysis by a Sona-Graph. The investigation indicated that the sibilant sounds produced the closest speaking space and that the mean and the variability of the closest speaking space in Cantonese speakers were smaller than that in English speakers. The beliefs that the closest speaking space was smaller than that of the freeway space and that the speaking space of m sound was similar to the freeway space were not supported by this study. PMID- 8227692 TI - Surface properties of elastomeric impression materials. AB - This study investigated the wettability of commonly used elastomeric impression materials and their gypsum castability. The wettability was evaluated by determining the contact angles of drops of a CaSO4 aqueous solution on flat impression surfaces. Gypsum castability was determined by counting the number of voids formed in die stone casts made from impressions of a critical die. All specimens used in this study were exposed to saliva to simulate clinical conditions, where materials spontaneously acquire salivary origin biofilms. The results indicate that the condensation silicones and conventional addition silicones exhibited the largest contact angles and the highest number of voids. The new generation of 'hydrophilic' addition silicones had higher wettability than the conventional silicones and an improved gypsum castability, comparable to that of the polyether materials. PMID- 8227693 TI - Two ceramic materials compared for the production of CEREC inlays. AB - Two ceramic materials, MGC Dicor and Vitablocs Mark II, were compared in vitro for the quality of fit of CEREC inlay restorations. The inlays made from Vitablocs Mark II fitted the inlay preparation significantly better than those made from MGC Dicor. PMID- 8227694 TI - Depth of cure of radiation-activated materials--effect of mould material and cavity size. AB - A method for determining depth of cure of radiation-activated restorative materials has been developed. The effect of mould material and cavity size on depth of cure was determined using a digital penetrometer method. Values were obtained for three selected shades of three dentine products and, in addition, the same three shades of an enamel variant were assessed. All these values were obtained using a 4 mm diameter x 6 mm deep stainless steel mould. PMID- 8227695 TI - Load transmission evaluation by removable distal-extension partial dentures using holographic interferometry. AB - Five pairs of bilateral distal-extension removable partial dentures with different clasp designs were constructed on the same dried human mandible in order to compare their load distribution characteristics. The technique of holographic interferometry was used. Among the dentures tested, those with RPI and back action clasps having mesial rests provided the best results from the mechanical point of view. The most unfavourable behaviour was presented by the Akers clasps design. The RPA design and dentures having clasps with continuous MOD rest gave intermediate results. PMID- 8227696 TI - The AADR, the dental industry, and their constituencies. PMID- 8227697 TI - Platelet-derived growth factor stimulates hyaluronate but not proteoglycan synthesis by human gingival fibroblasts in vitro. AB - The effect of PDGF-BB on human gingival fibroblasts was monitored in an in vitro system. PDGF was found to be mitogenic for these cells, although it required the presence of low concentrations of fetal calf serum to be active. Proteoglycan and hyaluronate synthesis was analyzed by labeling newly synthesized macromolecules with [35S]-sulfate or [3H]-glucosamine, respectively. Identification of specific glycosaminoglycans was achieved by selective enzymatic or chemical degradations. It was found that cells cultured in the presence of PDGF showed no discernible differences in proteoglycan synthesis relative to the control cultures. There were no alterations in amounts of proteoglycans synthesized, types of sulfated glycosaminoglycans synthesized, or relative hydrodynamic sizes of the proteoglycans. In contrast to the proteoglycans, hyaluronate synthesis was significantly increased in the presence of PDGF. The increase in [3H]-glucosamine incorporation into newly synthesized hyaluronate correlated with an increase in the activity of the enzyme hyaluronate synthetase but could not be accounted for entirely by changes in the specific activity of sugar nucleotide precursors, which did alter slightly under differing culture conditions. It is concluded from these results that PDGF stimulates gingival fibroblasts to proliferate and is associated with a differential effect of proteoglycan and hyaluronate synthesis. These observations may correlate with the observed early events associated with wound healing and repair. PMID- 8227698 TI - Involvement of the central nervous system in the salivary secretion induced by pilocarpine in rats. AB - The effect of rats of an anteroventral third ventricle (AV3V) electrolytic lesion on salivary secretion induced by intraperitoneal (i.p.) or intracerebroventricular (i.c.v.) injection of a cholinergic agonist (pilocarpine) was investigated. Sham- or AV3V-lesioned rats anesthetized with urethane and with a stainless steel cannula implanted into the lateral ventricle (LV) were used. The amount of salivary secretion was studied over a seven-minute period after i.c.v. or i.p. injection of pilocarpine. In sham-operated rats, i.p. injection of pilocarpine (1 mg/kg b.w.) (after 6 h, 2, 7, and 15 days) produced salivary secretion (486 +/- 21, 778 +/- 85, 630 +/- 50, and 560 +/- 55 mg/7 min, respectively). This effect was reduced 6 h, 2, and 7 days after an AV3V lesion (142 +/- 22, 113 +/- 32, and 290 +/- 62 mg/7 min, respectively), but not 15 days after an AV3V lesion (516 +/- 19 mg/7 min). I.c.v. injection of pilocarpine (120 micrograms in 1 microL), in sham-operated rats after 6 h, 2, 7, and 15 days also produced salivary secretion (443 +/- 20, 417 +/- 81, 496 +/- 14, and 427 +/- 47 mg/7 min, respectively). The effects of i.c.v. pilocarpine were also reduced 6 h, 2, and 7 days after an AV3V lesion (143 +/- 19, 273 +/- 14, and 322 +/- 17 mg/7 min, respectively), but not after 15 days (450 +/- 28 mg/7 min). The results demonstrate that the central nervous system, and particularly the AV3V region, is important for the effect of pilocarpine on salivary secretion in rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227699 TI - Motor unit territories and fiber types in rabbit masseter muscle. AB - The myosin heavy chain (MHC) content and spatial distribution of the fibers of 11 motor units (MUs) of the rabbit masseter muscle were determined. The fibers of single MUs were visualized in whole-muscle serial sections by a negative periodic acid/Schiff reaction for glycogen after they had been depleted of glycogen by extracellular stimulation of their motoneuron in the trigeminal motor nucleus. The MHC isoforms present in the fibers were characterized by monoclonal antibodies. Individual fibers appeared to contain from one to three MHC isoforms. In six cases, all fibers of a motor unit had an identical MHC content; in five cases, different fiber types were found in a single unit. The fiber number per MU varied between 40 and 424, the territory size between 1.1 and 11.0 mm2 (of a total muscle cross-section of 200 mm2), and fiber density between 6 and 17 MU fibers per 100 muscle fibers. In the multipennate masseter, the fibers were usually restricted to a single anatomical compartment. In comparison with leg muscles, the fibers of the masseter motor units, although similar in number, were restricted to relatively smaller subvolumes of the muscle and thus reached higher densities in their territories. The small territories are the anatomical substrate for the observed heterogeneity of motor behavior. Since the different anatomical compartments of the masseter differ with respect to their biomechanical capabilities, this makes this muscle multifunctional in the exertion of complex motor tasks. PMID- 8227700 TI - Co-activation of sternocleidomastoid muscles during maximum clenching. AB - In an attempt to determine the degree of co-activation present in selected cervical muscles during clenching, we instructed 12 male subjects to produce four brief maximum voluntary contraction (MVC) efforts (clenching) in a position of maximum intercuspation. Surface EMG activity was recorded bilaterally from the masseter and sternocleidomastoid (SCM) muscles. The contraction level for the SCM during clenching was reported as a percentage of the SCM's maximum activity achieved during maximum neck flexion against resistance. All EMG signals for the masseter and SCM were converted to a true RMS voltage signal and digitized at a 100-Hz sampling rate. Mean peak EMG voltage levels were determined for the activity recorded during each brief MVC task. All subjects demonstrated co activation of the SCM during strong abrupt clenching efforts. The mean levels (+/ S.D.) of SCM activity were 11.8 +/- 9.6% (right) and 14.2 +/- 9.4% (left) of the MVC capacity. Fifty percent of masseter activity was required to achieve 5% activity of the SCM bilaterally, and there was a progressive development of the SCM co-activation which paralleled the masseter activation. PMID- 8227701 TI - Sternocleidomastoid muscle inhibition induced by trigeminal stimulation. AB - Among numerous reports of anatomical and functional coupling between the trigeminal and cervical systems is the demonstration that the sternocleidomastoid (SCM) muscles may become activated along with the masseter muscles during forceful abrupt biting maneuvers. Whether the co-activated SCM is also inhibited by stimuli that produce masseter inhibition is not known. This study evaluated the SCM for the presence of inhibition during mechanically-elicited (chin or forehead tap) and electrically-elicited (anterior maxillary gingiva stimulation) inhibition of the masseter muscle in ten healthy men. Surface EMG data were recorded bilaterally from the masseter and SCM muscles. The data for each muscle were converted to ratios of the pre-stimulus maximum voluntary contraction activity for each subject and averaged across subjects. Means of these percentages were determined at several defined pre- and post-stimulus intervals. The results indicate that masseter inhibition was clearly elicited by the electrical and both forms of mechanical stimulation. SCM co-inhibition could be evoked by electrical and chin tap stimulation but not by forehead tap. The responses to these stimuli varied among subjects, from trial to trial, and within subjects depending on the experimental condition. The fact that it was possible for this co-inhibition to be evoked is presented as further indication of the functional coupling of the trigeminal and cervical systems. PMID- 8227702 TI - Prevalence in the Dutch adult population and a meta-analysis of signs and symptoms of temporomandibular disorder. AB - A nationwide survey of oral conditions, treatment needs, and attitudes toward dental health care in Dutch adults was carried out in 1986. One of the aims of the study was to assess the prevalence of signs and symptoms of temporomandibular disorder (TMD). A sample of 6577 persons (from 15 to 74 yr of age), stratified for gender, age, region, and socio-economic status, was contacted. Of this sample, 4496 persons participated in the behavioral part of the study, of whom 3526 were examined clinically. The TMD prevalence was based on (1) perceived signs and symptoms of TMD and (2) clinical examination of joint sounds, deviation, and pain on mandibular movements. A total of 21.5% of the Dutch adult population perceived some dysfunction, and 44.4% showed clinically assessed signs and symptoms of TMD. In nearly all age groups, the signs and symptoms of TMD appeared more in women than in men. Agreement between the results of the clinical examination and the anamnestic dysfunction index was significant (p < 0.0001); however, the Pearson's correlation coefficient was low (r = 0.29). The odds-value (risk-ratio) that subjects who perceived signs and symptoms of TMD would present with clinically assessed signs and symptoms of TMD was 2.3. The results of the survey were compared with results of a meta-analysis performed on 51 TMD prevalence studies. The analysis revealed (1) a perceived dysfunction rate of 30% and (2) a clinically assessed dysfunction of 44%, both based on compound samples of, respectively, over 15,000 (23 studies) and over 16,000 (22 studies) randomly selected subjects. PMID- 8227703 TI - Defluoridation of water at high pH with use of brushite, calcium hydroxide, and bone char. AB - The aim of this study was to improve the efficiency of the bone-char method of water defluoridation by pre-treating the water with brushite and calcium hydroxide. Various amounts of brushite, calcium hydroxide, and bone char were suspended batchwise in 100 mL of distilled water containing 0.53 mmol/L fluoride for 24 h under gentle agitation. At suitable intervals, pH and the concentrations of fluoride, calcium, and phosphate in the water were determined and, when possible, the degrees of saturation with respect to brushite, hydroxyapatite, and fluorapatite calculated. Bone char used alone took up fluoride slowly and inefficiently. The addition of brushite and calcium hydroxide resulted in high concentrations of calcium and phosphate, making the solutions highly supersaturated with respect to fluorapatite, and led to a 20-fold increase in fluoride removal from the water. The combined use of all three salts left low concentrations of phosphate in solution and optimized the fluoride uptake capacity. Repeated use of the same bone char for 18 consecutive runs demonstrated that uptake of fluoride by the bone char was improved by repeated use, provided that brushite and calcium hydroxide were added. Therefore, addition of the two salts to the water may prolong the life of the bone char indefinitely, ensure the removal of fluoride, and thus avoid the problem of determining when the bone char is exhausted. In conclusion, we show that the bone-char defluoridation technique can be improved by addition of brushite and calcium hydroxide to the water. The problem of high terminal pH remains, however, and further work is required to improve potability. PMID- 8227704 TI - The effects of enamel anisotropy on the distribution of stress in a tooth. AB - Enamel is thought to have highly anisotropic stiffness characteristics, because of its prismatic structure. It is probable that the enamel is stiffer in the prism direction compared with a direction perpendicular to it. The prisms are thought to run approximately perpendicular to the enamel-dentin junction. The curvilinear anisotropy that will result can readily be modeled by TOMECH, a finite element program developed at the University of Sheffield, since curvilinearity of mechanical properties is available as an automated feature of this program. The patterns of stress due to an external load were investigated in two-dimensional abstract models, and in a model of a mandibular second premolar, for both anisotropic and isotropic enamel. Results were compared with the commercial code ANSYS and good agreement obtained. Enamel with anisotropic properties was found to have a profoundly different stress distribution under load when compared with models with isotropic enamel. For isotropic enamel, the load path is directed through the stiff enamel shell, while for anisotropic enamel, the load path is directed into the dentin, as the load path follows the stiff direction of the enamel prisms. Thus, if enamel is indeed anisotropic, its function differs greatly from that suggested in previous hypotheses. Enamel with anisotropic material characteristics would provide a hard-wearing protective surface-coating while simultaneously diverting the load away from this brittle, low-tensile-strength phase, thus reducing the potential for tooth fracture. PMID- 8227705 TI - Interferometric measurements of cusp deformation of teeth restored with composites. AB - A Michelson interferometry apparatus was used for measurement of the displacement of the buccal cusps of premolars after restoration of MOD preparations with composites. The effects of composite type, cavity size, and hydration conditions were examined. Interferometry permitted real-time measurement of cusp movement as it occurred. Contraction occurred very rapidly, about 1/3 of the 60-minute amount within the two-minute period of exposure to the curing light. Cusp movement was smooth rather than interrupted, indicating lack of microfracturing at deformations of 11-46 microns. Contraction, 0.94% for Heliomolar and 1.2% for P 50, was similar to the linear polymerization shrinkage of the resins. Less cusp movement occurred in small cavities than in large cavities. Hydrated teeth had less cusp movement than dehydrated teeth. PMID- 8227706 TI - Distributed crack analysis of ceramic inlays. AB - In all-ceramic restorations, crack formation and propagation phenomena are of major concern, since they may result in intra-oral fracture. The objective of this study was calculation of damage in porcelain MOD inlays by utilization of a finite-element (FE) implementation of the distributed crack theory. "Damage" is defined as the parameter that describes the local decrease of stiffness caused by microdefects. In the simulated MOD ceramic inlay, the crack initiation starts at the internal occlusal surface near the pulpo-axial line angle. This initiation is invisible from the external surface and cannot be detected by the clinician. The crack initiation at the internal surface started as soon as 55-60% of the loading needed for complete fracture was reached. The use of FE techniques for calculation of the fracture in loaded ceramic inlays offers prospects for further detailed study of the crack behavior, including three-dimensional modeling and cyclic loading situations. PMID- 8227707 TI - Active specific immunotherapy of malignant melanoma with anti-idiotypic monoclonal antibodies. AB - In this study, we have utilized the human high molecular weight-melanoma associated antigen (HMW-MAA) as a target for active specific immunotherapy with mouse anti-idiotypic (anti-id) monoclonal antibodies (mAb) in patients with malignant melanoma. After having summarized the characteristics of HMW-MAA which account for its selection as a target for immunotherapy, we describe the development and characterization of mouse anti-id mAb MK2-23 which bears the internal image of HMW-MAA. Furthermore, we describe the results of the first clinical trial performed with mouse anti-id mAb MK2-23 in patients with malignant melanoma. PMID- 8227708 TI - The relationship among minimal erythema dose, minimal delayed tanning dose, and skin color. AB - The relationship among minimal erythema dose (MED), minimal delayed tanning dose (MDTD), and skin color was examined in 16 healthy volunteers using three different spectra. The subjects were exposed to UVB, UVA+B, and UV+Visible light (UV+Visible) with a xenon arc solar simulator as a light source. The MEDs for UVB and UVA+B were less than the MDTDs, whereas the MED for UV+Visible was higher than the MDTD. There was no significant correlation between the MED and the MDTD for UVB or UVA+B. The MED for UV+Visible was significantly correlated to the MDTD (p < 0.01). Skin color significantly correlated with MEDs for UVB and UVA+B (p < 0.01), but not for UV+Visible. There was no significant correlation between skin color and the MDTD for any spectra. From these results, it is suggested that the relationship between erythemal and melanogenic responses is dependent on spectral bands of the light source and that skin color is a predictor of UV-induced erythema. PMID- 8227709 TI - Experimental murine protoporphyria induced by griseofulvin (GF): the relationship between hepatic porphyrin levels and liver function test values in mice treated with GF. AB - To investigate the hepatic abnormalities accompanying experimental protoporphyria due to griseofulvin (GF), liver function test values and porphyrin levels in mice were assayed at days 2, 4, 8, and 16 after starting the administration of 0.5% GF feed. Furthermore, in an attempt to elucidate the harmful effects of GF on liver functions, the above mentioned assay was also performed after the feed was discontinued in mice given 0.5% GF feed for 16 days. The hepatic protoporphyrin (PP) level had already risen by day 2, but the erythrocytic PP level was within normal limits at that time. Hepatic PP levels increased gradually, followed by an increase in erythrocytic PP levels. The variation in liver function test values roughly paralleled the porphyrin levels. Over the time span of the response to GF, the variations in the serum glutamate oxaloacetate transaminase (S-GOT) levels, serum glutamate pyruvate transaminase (S-GPT) levels, and leucine amino peptidase (LAP) levels resembled those in hepatic PP. On the other hand, the changes in alkaline phosphatase (ALP) levels paralleled those of the erythrocytic PP levels. Erythrocytic and fecal protoporphyrin levels decreased to the normal level one month after the discontinuation of GF administration, but the hepatic protoporphyrin level still was 53.6 times higher than the normal level two months after switching to normal feed. The values of liver function tests had returned to within the normal range after one month. By the fourth day after the administration of GF, a brown pigmented material could be observed around the hepatocytes and the Glisson sheath; the amount of this material increased day by day.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227710 TI - A case of cystic dermal duct tumor corresponding to poroid hidradenoma. AB - A case of dermal duct tumor on the back of a 61-year-old male, representing a cystic lesion, is reported. Such cystic dermal duct tumors seem to be considered poroid hidradenomas according to the recently suggested classification of poromas. Poroid hidradenoma is rarely mentioned in the literature, and no description of its clinical features has yet been published. Carcinoembryonic antigen (CEA) immunostaining was positive in the tumor cells adjacent to the cystic space. We suppose that these CEA-positive cells were associated with cyst formation in our case. PMID- 8227711 TI - Calcinosis cutis in juvenile dermatomyositis responsive to aluminum hydroxide treatment. AB - Calcinosis cutis is a frequent complication of juvenile dermatomyositis; however, its treatment remains unsatisfactory. We report a case of calcinosis cutis in juvenile dermatomyositis which was successfully treated with oral aluminum hydroxide. Almost complete clearing of calcinosis was observed after 8 months of therapy. There were no adverse effects from therapy. Aluminum hydroxide is worth trying in treating calcinosis cutis. PMID- 8227712 TI - Acquired dermal melanocytosis of the hand: a new clinical type of dermal melanocytosis. AB - We report a 22-year-old man with acquired dermal melanocytosis on the hand which developed without any history of previous inflammation. He had no history of contact with or oral administration of any drugs or metals which might have caused pigmentation. Histopathologically, spindle-shaped dermal melanocytes were observed running parallel to the collagen bundles and scattered in all portions of the dermis. The dopa reaction of these cells was positive. Ultrastructurally, dermal melanocytes containing numerous melanosomes (stages II to IV) which were surrounded by the extracellular sheath were observed. Differential diagnosis of other dermal melanocytosis was discussed. PMID- 8227713 TI - A case of lupus meningitis treated successfully with methylprednisolone pulse therapy. AB - A 46-year-old female had suffered from systemic lupus erythematosus (SLE) for 8 years. Headache, vomiting and stiff neck appeared in the active phase of SLE. Findings in the cerebrospinal fluid were consistent with those of lupus meningitis. No pathogenic microbes were detected by microbiological or immunological examinations. She was diagnosed as having lupus meningitis. The method discussed herein which elucidates the cause of fever in SLE using white blood cell count (WBC) and alpha-2 globulin appeared to be useful for examining this case of meningitis. Lupus meningitis seems to preferentially occur in SLE patients with positive anti-ribonucleoprotein (RNP) antibody. Pulse therapy with methylprednisolone appeared to work well in this lupus meningitis patient who had had a long course of corticosteroid therapy. PMID- 8227714 TI - A case of acrogeria. AB - We report a 30-year-old Japanese male with acrogeria. He was in good health except for the prominent skin atrophy over the hands and feet. In the microscopic view of an atrophic skin biopsy specimen, elastic fibers were clumped and fragmented, especially in the upper dermis. Reviewing fourteen previously reported Japanese cases of acrogeria, the decrease and degeneration of elastic fibers were the prominent characteristic, as was the atrophy of subcutaneous fat. PMID- 8227715 TI - Isolated lichen planus of the nails treated with etretinate. AB - A 46-year-old Japanese man with isolated lichen planus of the nails is reported. The affected nails were all of the fingernails and the third and fourth toenails of both feet. Histologically, the ventral part of the proximal nail folds showed mild band-like cell infiltration below the epidermis, and the nail beds (ventral matrix) showed hypergranulosis and epidermal-dermal interphase activity with liquefaction degeneration of basal cells and a band-like cell infiltration consisting of lymphoid cells and histiocytes. Systemic use of etretinate in combination with application of steroid lotion was evaluated as very effective. Although the use of retinoids for lichen planus of the nails remains only a suggested method of treatment, a positive trial should be attempted, since this disorder sometimes results in severe nail damage. PMID- 8227716 TI - 1 alpha,25-dihydroxy vitamin D3 modulation of HLA-DR mRNA induced by gamma interferon in cultured epithelial tumor cell lines. AB - Using three cultured epithelial tumor cell lines, we investigated and analyzed the effects of gamma-interferon (gamma-IFN) and 1 alpha,25-dihydroxy vitamin D3 (1,25-(OH)2D3) on the levels of HLA-DR (alpha) mRNA and HLA-DR (beta) mRNA by Northern blot analysis. After treatment with gamma-IFN alone, the levels of the mRNA increased. Treatment with both gamma-IFN and 1,25-(OH)2D3 at the same time resulted in a significant decrease in the levels of mRNA in K-TL-1, IK-TL-2, and M-TL cells as compared to those induced by gamma-IFN alone. PMID- 8227717 TI - Analysis of the interleukin 6 receptor on normal human keratinocytes by digital imaging fluorescence microscopy. AB - The interleukin 6 receptor (IL 6 receptor) on normal human keratinocytes was analyzed by digital imaging fluorescence microscopy (DIFM). Non-fixed keratinocytes were incubated with monoclonal anti-IL 6 receptor antibody (MT18). The fluorescence intensity was quantitatively determined by an interactive laser cytometer (ACAS 470). IL 6 receptor-positive cells showed high density staining with the specific antibody in the center of the elevated cell surface. Therefore, DIFM appears to be an effective method for study of the distribution of IL 6 receptors at the single living cell level. PMID- 8227718 TI - Elm Road and Hillsborough: tragedy, the law and medicine. PMID- 8227719 TI - Neuromuscular abnormalities in critically ill patients. PMID- 8227720 TI - Epidemiological markers of coagulase-negative staphylococci. AB - Several different epidemiological typing methods have been used in studies of coagulase-negative staphylococci; these include biotyping, antibiotic susceptibility pattern analysis, serological typing, phage typing, slime production detection, protein profile analysis, immunoblot fingerprinting and DNA typing. The most frequently used tests are antibiotic susceptibility and extrachromosomal DNA banding patterns. Comparison of chromosomal DNA restriction patterns is facilitated when the fragments are separated by pulsed-field electrophoresis. Typing variations occur quite frequently even in isolates of demonstrated clinical significance. Caution should therefore be exercised in denying the clinical relevance of coagulase-negative staphylococci, even when successive isolates show distinct typing differences. PMID- 8227721 TI - Trends from the United States with end of life decisions in the intensive care unit. AB - OBJECTIVE: To describe the changes that have occurred in the United States since medicine has moved away from a paternalistic model to one that promotes patient autonomy and self-determination. To discuss the implications for cardiopulmonary resuscitation (CPR) and the increasing use of when not to perform CPR and other life-sustaining therapies. To describe the various interpretations of the ritual term Do-Not-Resuscitate (DNR) and to introduce the concept of futility in the context of non-beneficial over-treatment and discriminatory under-treatment. SETTING: Selected clinical, philosophical and public policy literature and two illustrative case examples. RESULTS: 1. There is no longer a mandate to perform CPR on all dying patients, even though the Council on Ethical and Judicial Affairs of the American Medical Association in 1991 said that the only restrictions should be in patients with an irreversible terminal condition or when the physician writes the order, DNR. 2. The DNR order usually requires the informed refusal of CPR by the patient or family. There is only minimal support for a unilateral decision even for patients with far advanced disease. 3. DNR is often the first step in the negotiated process of forgoing care in the ICU. There are multiple interpretations of DNR both in and outside of the ICU. 4. Health Proxy is the latest attempt to have a person clarify his/her wishes and preferences by naming a decision maker, if the individual losses mental capacity. 5. Although ethical principles seem well established, there are inconsistent interpretations and practices at the bedside in the United States in part due to the restructuring of the relationship between physicians and patients, providers and consumers/clients. 6. Objective severity scores such as Apache III, SAPS II, MPM II are generally not applicable for individual patient end-of-life decisions. CONCLUSIONS: Although Health Proxy in its current formulation has been disappointing, there is a clear trend for wider application of DNR and for more active discussions about withholding or forgoing other life-sustaining therapies. DNR has a different interpretation late into the ICU course (> 72 h) than when applied at or shortly after ICU admission. Late in the ICU course, it has been decided by the medical team and family or surrogate decision maker/Health Proxy that the patient has failed or is in the process of failing aggressive ICU therapy. Early use of DNR may be related to limitations based on pre-existing chronic or subacute disease burden or an unwillingness to proceed with a full ICU course of therapy. It is unclear how Ethics Committees, risk management and hospital administrators, national practice guidelines, governmental sponsored health care reform will interface with the highly complex individual patient- physician--family--Health Proxy interface as practiced in the United States. Dialogue between the Society of Critical Care Medicine and the European Society of Critical Care Medicine and among interested physicians could provide a format for a multi-cultural context to discuss end of life issues in the ICU setting. PMID- 8227722 TI - Anticoagulant regimens in acute continuous hemodiafiltration: a comparative study. AB - OBJECTIVE: To compare and contrast different heparin regimens for extracorporeal circuit anticoagulation in patients receiving acute continuous hemodiafiltration (ACHD). DESIGN: Prospective controlled randomized comparisons of the following regimens: 1) Low dose (500 IU/h) pre-filter heparin versus regional anticoagulation in patients on continuous arteriovenous hemodiafiltration (CAVHD) via A-V shunt. 2) Low dose pre-filter heparin versus no anticoagulation in patients receiving CAVHD via femoral cannulae. 3) Low dose pre-filter heparin versus regional anticoagulation in patients on continuous veno-venous hemodiafiltration (CVVHD). 4) An assessment of the consequences of the use of no anticoagulant in patients predicted to be at high risk of hemorrhagic complications on treatment with CVVHD. SETTING: University Teaching Hospital ICU. PATIENTS: 64 ICU patients with acute renal failure. MEASUREMENTS AND MAIN RESULTS: Haemofilter survival during shunt CAVHD was significantly prolonged by the use of regional anticoagulation compared to the use of low dose heparin (mean filter survival: 57.1 h versus 42.9 h; p < 0.05). In CAVHD using femoral cannulae, no significant differences in haemofilter survival were found between anticoagulation with low dose heparin and the use of no anticoagulant (mean filter survival: 55 h versus 52.5 h; NS). During CVVHD, regional anticoagulation compared to low dose heparin produced a trend towards prolonged filter life which was, however, not statistically significant (mean filter survival: 40.5 h versus 31.4 h; NS). In patients assessed to be at high risk of bleeding, CVVHD without anticoagulation provided a mean filter survival of 40.9 h (95% CI 27-54.8 h). CONCLUSIONS: Regional anticoagulation leads to longer filter survival than low dose heparin in shunt CAVHD. A regimen of no anticoagulation during femoral CAVHD leads to a filter life similar to that of low dose heparinization. During CVVHD, regional anticoagulation and low dose heparin are associated with similar filter survival times. In patients assessed to be at high risk of bleeding, treatment with CVVHD without anticoagulation results in adequate filter survival. PMID- 8227723 TI - Effects of ketoprofen on respiratory and circulatory changes in endotoxic shock. AB - OBJECTIVE: To study the effects of ketoprofen, a dual inhibitor of the arachidonic acid metabolism, on hemodynamic and respiratory changes during endotoxic shock. DESIGN: Prospective, randomised, controlled study using an established intact animal model of endotoxic shock in sheep. SETTING: An animal laboratory in a university hospital. INTERVENTIONS: 4 groups were studied (n = 7 in each). Group K received ketoprofen and group A received aspirin 30 min before start of endotoxin infusion. Group E received endotoxin, but no drug treatment. Group C received neither endotoxin nor drug treatment. All the animals were anaesthetised with ketamine, had controlled ventilation with FiO2 = 0.5 and received Ringer's lactate at an infusion rate that would keep the pulmonary capillary wedge pressure constant. RESULTS: Both ketoprofen and aspirin prevented the early rise in pulmonary arterial pressure that occurred in group E a few minutes after start of i.v. infusion of endotoxin. Furthermore, ketoprofen prevented any significant changes in arterial blood pressure, arterial oxygen tension, oxygen delivery index, oxygen extraction ratio, respiratory compliance, intrapulmonary shunt fraction, and platelet counts that occurred in group E. Aspirin, on the other hand, provided only partial and time limited (1-2 h) protection against these changes. Wet-to-dry weight ratios of the lungs were significantly lower in the ketoprofen treated than in the untreated shock controls and the aspirin treated animals. CONCLUSION: Ketoprofen completely prevented the changes in hemodynamics and respiratory function observed in control-endotoxin-treated animals. PMID- 8227724 TI - Prospective evaluation of self-extubations in a medical intensive care unit. AB - OBJECTIVE: To evaluate the incidence, associated factors and gravity of self extubations. DESIGN: Prospective study about all patients intubated over an 8 month period. SETTING: A medical intensive care unit of a University Hospital. PATIENTS: Patients were divided into two groups: self-extubated and those that did not. The self-extubations were separated into deliberate acts by the patients and accidental. RESULTS: 24 of the 197 patients included presented a total of 27 extubations (12%). There were 21 deliberate incidents and 6 accidental. The only differences between the cases and the rest of the population were a higher mean age (67 vs 59 years) and a larger proportion of chronic respiratory failure (66% versus 35%). Reintubation was necessary in 20 cases (74%) within 30 min in 16 cases. The main indication for reintubation was acute respiratory distress (90%). Reintubation was associated with one death. CONCLUSION: Self-extubation is a frequent and serious complication of mechanical ventilation. Deliberate self extubation, the most frequent type of incident could possibly be reduced by better sedation of agitated patients and accidental self-extubation by better training of the nursing staff. PMID- 8227725 TI - Noise in the ICU. AB - OBJECTIVE: The growing number of technical devices in ICUs makes noise exposure a major stressor. The purpose of this study was to assess noise levels during routine operation in our ICU. DESIGN: Our ICU is an open ward with four rooms, constructed in the 1960s. During the study period, 4 patients were in the controlled room and were treated by 4 nurses during the day and by 2 at night. A weighted sound pressure levels (SPL) were measured continuously for 2 days and nights. Also measured were the alarms of various appliances. For gross overall evaluation it is customary to state the Leq, i.e. the energy-averaged level during measurement. The annoyance caused by noise depends more on rare events of high intensity. Therefore, the distribution of SPL values (Ln) over time was also analysed. RESULTS: SPL was roughly the same during the day and at night, with Leq between 60-65 dB(A) and peaks up to 96 dB(A). Most alarms reach an SPL of 60-70 dB(A), but some exceed 80 dB(A). During teaching rounds Leq exceeds 65 dB(A). CONCLUSION: During the day and at night SPL always surpasses the permissible noise exposure for 24 h of 45 db(A) recommended by the US Environmental Protection Agency. Alarms cause the most irritating noise. Hospital management should pay attention to internal noise, and SPL should be measured routinely. PMID- 8227726 TI - Pharmacokinetics of vancomycin during continuous hemodiafiltration. AB - OBJECTIVE: To study the pharmacokinetics of vancomycin in three patients with acute renal failure related to multi-organ failure during continuous venovenous hemodiafiltration (CVVHD). DESIGN: Prospective exploratory, open-labelled study. SETTING: Critical Care Unit in a University Medical Centre. PATIENTS: 3 patients exhibiting hemodynamic instability and oligo-anuric acute renal failure requiring extra-renal epuration were included in this study. INTERVENTION: Every patient received 7.5 mg/kg IV vancomycin over 1 h for a documented or suspected nosocomial staphylococcal infection. Serum and dialysate outlets samples were collected before infusion and 1, 3, 6, 12, 18, 24 after the end of infusion. MEASUREMENTS AND RESULTS: Mean age was 58.7 years (range 41-79) and mean SAPS 15.7 (9-23). The mean peak concentrations were 27.3 mg/l (range 15.6-45.6) one hour after the end of infusion. The average remaining vancomycin concentration 24 h after the onset of infusion was 3.6 mg/l (range 2.6-4.5). The mean terminal disposition rate constant and elimination half-life were 0.05 h-1 and 13.9 h respectively. Mean total body clearance was 38.9 +/- 4.3 ml/min and dialysate outlet (DO) clearance 4.2 +/- 1.3 ml/min. The mean volume of distribution was 47.4 +/- 6.4 l. CONCLUSION: CVVHD is effective for vancomycin elimination. In these patients, the elimination half-life is almost constant, involving a following injection of vancomycin 12 h later to achieve effective concentrations. PMID- 8227727 TI - Performance of a hydrophobic heat and moisture exchanger at different ambient temperatures. AB - OBJECTIVE: To evaluate the effect of different room temperatures on hydrophobic heat and moisture exchangers (HME) humidifying capability and efficiency. METHODS: Stock HMEs were tested in vitro using an already described test model, with separation of inspiratory and expiratory gas. Absolute humidity (AH) was measured by means of dry-wet dual thermocouple, and HME efficiency was computed as the ratio between expired to inspired AH, at room temperature of 20 and 26 degrees C. RESULTS: Inspired gas temperature and AH were significantly higher at 26 than at 20 degrees C; since expired AH remained substantially unchanged, HME efficiency was also higher in warmer environment. CONCLUSIONS: Hydrophobic HMEs appear to be affected by room temperature, increasing their humidifying ability and their efficiency with its rise. PMID- 8227728 TI - Intensive use of general ultrasound in the intensive care unit. Prospective study of 150 consecutive patients. AB - OBJECTIVE: To assess the impact of systematic ultrasound (US) examination in patients admitted to the ICU. DESIGN: An observational study of 150 consecutive patients. SETTING: A medical ICU of a University-affiliated hospital. PATIENTS: All consecutively admitted patients were examined, but patients discharged within < 48 h were excluded from the analysis. INTERVENTIONS: A systematic examination of the abdomen, pleural space, cervical and femoral vein, was performed at the bedside by the same investigator. MEASUREMENTS: Results of US examinations were compared to findings from CT-scan, surgery, endoscopic procedures, autopsy, or other diagnostic tests. The impact of U.S. findings on immediate patients management was evaluated. RESULTS: In 33 of 150 patients (22%), U.S. findings influenced the diagnosis, work up of the patients, and had a direct impact on the therapeutic plan. In 30/33 cases, confirmation of diagnosis was obtained by another diagnostic and/or therapeutic procedure. CONCLUSIONS: Routine US examination may alter therapeutic plans in up to 1/4 of critically ill patients admitted to the ICU. US examination should be liberally performed on such patients. PMID- 8227729 TI - Phaeochromocytoma presenting as acute circulatory collapse and abdominal pain. AB - Phaeochromocytomas may present as an acute emergency with a perplexing variety of symptoms. We report a case which presented with abdominal pain and severe respiratory distress due to a ruptured haemorrhagic phaeochromocytoma. The severe cardiorespiratory collapse which precipitated admission to the intensive care unit was managed with high doses of dobutamine and noradrenaline and the patient was stable during the operative resection. The management of this previously undiagnosed phaeochromocytoma and its atypical presentation are discussed. PMID- 8227730 TI - Multiple organ failure after mitral valve repair with intravascular hemolysis and its recovery due to mitral valve replacement. AB - We present a patient with evidence of severe intravascular hemolysis after mitral valve repair, an established method for the surgical treatment of mitral valve disease to avoid prosthesis related complications. The coincidence of this uncommon complication with hemodynamic instability due to pre-existing myocardial dysfunction and Gram-negative pneumonia promoted the development of simultaneous dysfunction of liver, kidney and the cardio-respiratory system. Elimination of the source of hemolysis by re-operation with mitral valve replacement on the ninth postoperative day allowed prompt recovery from severe organ dysfunction. Free hemoglobin may have perpetuated progressive organ failure in our patient. PMID- 8227731 TI - Post-operative acute respiratory failure associated with Mycobacterium xenopi. PMID- 8227732 TI - Association between plasma ionized calcium and lactate concentration. PMID- 8227733 TI - Fat embolism in a patient with rhabdomyolysis: a rare cause of ARDS? PMID- 8227734 TI - Inhibitor substitution in sepsis. PMID- 8227735 TI - AT III in septicemia with DIC. AB - In the development of sepsis DIC is a common complication. Several studies presented in this paper show a coincidence between the development of DIC and depletion of Antithrombin III, a serine protease inhibitor which inhibits a large scale of activated clotting factors. It seems very probable that substitution therapy should be of benefit in the treatment of sepsis-related DIC and may improve the outcome of septic patients. Physiological and clinical findings are put together to clarify the basic rationale for running clinical trials and future studies. PMID- 8227736 TI - Perspectives for the future. AB - The pathophysiology of sepsis and septic shock is extremely complex and ultimately involves every physiological pathway. The initiating event is the entrance of endotoxin or similar substances into the blood which initiates the release of multiple mediators. These are designed to react locally and to protect the organism. Their constant release, however, sets in motion up- and down regulations, ultimately resulting in "metabolic anarchy". Tumor necrosis factor alpha and other cytokines trigger several systems, especially coagulation to yield DIC, and the complement system. Many treatment modalities have been developed, most recently those which substitute inhibitors of various systems. Antithrombin III concentrates and potentially protein C concentrates are designed to arrest DIC. C1-esterase inhibitor concentrates should intercept the activation of the complement system and the contact phase of coagulation and its relationship to kinin generation. Even newer approaches entail antibodies to tumor necrosis factor alpha or endotoxin itself. The complex process of sepsis will undoubtedly require a multifaceted therapeutic approach. PMID- 8227737 TI - New biological concepts on coagulation inhibitors. PMID- 8227738 TI - Coagulation disorders in septic shock. AB - Abnormalities in coagulation and fibrinolysis are frequently observed in septic shock. The most pronounced clinical manifestation is disseminated intravascular coagulation. Recent studies in human volunteers and animal models have clarified the early dynamics and route of activation of both coagulation and fibrinolytic pathways. In healthy subjects subjected to a low dose of either endotoxin or TNF an imbalance in the procoagulant and the fibrinolytic mechanisms is apparent, resulting in a procoagulant state. Also in patients with septic shock a dynamic process of coagulation and fibrinolysis is ongoing with evidence of impaired fibrinolysis. These abnormalities have prognostic significance; the extent of disturbances of coagulation and fibrinolysis is related to the development of multiple organ failure and death. PMID- 8227739 TI - Superdirective arrays for hearing aids. AB - Microphone arrays are the most effective of the techniques that have been proposed for improving speech intelligibility in noise for the hearing impaired. However, classical delay-and-sum beamforming provides very small amounts of array gain at low frequencies, while adaptive array processing has been shown to cancel the desired signal in the presence of strong room reflections. Superdirective arrays offer a heretofore overlooked solution in which optimal performance can be obtained for a stationary random noise field, but where the desired signal will not be canceled. A short constrained superdirective array suitable for hearing aid applications is proposed in this paper, and its theoretical performance is evaluated. PMID- 8227740 TI - Quantitative relationship between tissue composition and scattering of ultrasound. AB - This study establishes quantitative laws by which the magnitude of ultrasonic scatter is related to the gross chemical composition of tissues, viz., water, fat, and collagen. Tissues are modeled as immiscible mixtures, the constituents of which are uniformly dispersed over the volume. Each component of the mixture causes the ultrasonic velocity of the medium to fluctuate about the mean value. Such localized variations in the refractive index of a medium cause propagating sound waves to scatter. It is proposed that the contribution of each component to the total fluctuation is in proportion to its volume fraction. The analysis shows that the scatter of ultrasound exhibits a complex nonlinear dependence on the concentrations of the individual components. From the knowledge of sound speed of individual components, total velocity fluctuations are determined for binary and three-component models of the tissue. Comparison of these values, with those estimated from scatter measurements, shows a close correlation between the two sets. Collagen, although present in relatively small amounts in soft tissues, contributes a significant portion of the total scatter. However, both fat and collagen fractions must be taken into consideration to account for the major fraction of the observed scatter. In conclusion, there is a direct relationship between tissue composition and scatter from tissues. Such dependence must also be taken into consideration, in addition to the correlation function and correlation lengths to explain backscatter or loss in a transmitted signal due to scatter from tissues. PMID- 8227741 TI - Spectral-shape features versus formants as acoustic correlates for vowels. AB - The first three formants, i.e., the first three spectral prominences of the short time magnitude spectra, have been the most commonly used acoustic cues for vowels ever since the work of Peterson and Barney [J. Acoust. Soc. Am. 24, 175-184 (1952)]. However, spectral shape features, which encode the global smoothed spectrum, provide a more complete spectral description, and therefore might be even better acoustic correlates for vowels. In this study automatic vowel classification experiments were used to compare formants and spectral-shape features for monopthongal vowels spoken in the context of isolated CVC words, under a variety of conditions. The roles of static and time-varying information for vowel discrimination were also compared. Spectral shape was encoded using the coefficients in a cosine expansion of the nonlinearly scaled magnitude spectrum. Under almost all conditions investigated, in the absence of fundamental frequency (F0) information, automatic vowel classification based on spectral-shape features was superior to that based on formants. If F0 was used as an additional feature, vowel classification based on spectral shape features was still superior to that based on formants, but the differences between the two feature sets were reduced. It was also found that the error pattern of perceptual confusions was more closely correlated with errors in automatic classification obtained from spectral shape features than with classification errors from formants. Therefore it is concluded that spectral-shape features are a more complete set of acoustic correlates for vowel identity than are formants. In comparing static and time varying features, static features were the most important for vowel discrimination, but feature trajectories were valuable secondary sources of information. PMID- 8227742 TI - On the intonation of sinusoidal sentences: contour and pitch height. AB - A sinusoidal replica of a sentence evokes a clear impression of intonation despite the absence of the primary acoustic correlate of intonation, the fundamental frequency. Our previous studies employed a test of differential similarity to determine that the tone analog of the first formant is a probable acoustic correlate of sinusoidal sentence intonation. Though the typical acoustic and perceptual effects of the fundamental frequency and the first formant differ greatly, our finding was anticipated by reports that harmonics of the fundamental within the dominance region provide the basis for impressions of pitch more generally. The frequency extent of the dominance region roughly matches the range of variability typical of the first formant. Here, we report two additional tests with sinusoidal replicas to identify the relevant physical attributes of the first formant analog that figure in the perception of intonation. These experiments determined (1) that listeners represent sinusoidal intonation as a pattern of relative pitch changes correlated with the frequency of the tonal replica of the first formant, and (2) that sinusoidal sentence intonation is probably a close match to the pitch height of the first formant tone. These findings show that some aspects of auditory pitch perception apply to the perception of intonation; and, that impressions of pitch of a multicomponent nonharmonic signal can be derived from the component within the dominance region. PMID- 8227743 TI - Objective analysis versus subjective assessment of vowels pronounced by native, non-native, and deaf male speakers of Dutch. AB - An objective analysis has been performed on all 15 Dutch vowels pronounced in /hVt/ words by nine native Dutch, nine non-native, and six deaf males. Spectral representations of the vowel segments were created by determining the mean output levels of a bank of 16 filters (90-7200 Hz), with 1/3-oct bandwidths and logarithmic spacing of their center frequencies. The adequacy of the objective analysis is determined by the extent to which spectral information provides an accurate description of pronunciation quality. Spectral distances between the 24 utterances of each monophthong agree rather well with subjective distances obtained by listeners in an elaborate paired-comparisons experiment. For the various monophthongs, the correlation coefficients are within the range 0.63 to 0.88; averaging across all 12 monophthongs of each speaker results in a coefficient of 0.94. Furthermore, it appeared that the objective spectral analysis is as reliable as a subjective assessment by magnitude estimation by two to three listeners. Using principal components analysis (PCA), the number of dimensions by which the vowel spectra are described can be reduced. For the various monophthongs the range of the correlation coefficients between subjective distances and objective distances in a two-dimensional PCA subspace is 0.30-0.93. The three groups of speakers can still be distinguished in this subspace. In the extreme case of the deaf speakers all vowels are strongly "neutralized," whereas the different vowels of the native speakers are well separated, especially after speaker normalization; results are less clear for the non-natives. PMID- 8227744 TI - Effect of relative amplitude of frication on perception of place of articulation. AB - The amplitude of frication relative to vowel onset amplitude in the F3 and F5 formant frequency regions was manipulated for the synthetic fricative contrasts /s/-/integral of/ and /s/-/theta/, respectively. The influence of this relative amplitude manipulation on listeners' perception of place of articulation was tested by (1) varying the duration of frication from 30 to 140 ms, (2) pairing the frication noise with different vowels /i a u/, (3) placing formant transitions in conflict with relative amplitude, and (4) holding relative amplitude constant within a continuum while varying formant transitions and the amplitudes of spectral regions where relative amplitude was not manipulated. To determine if listeners were using absolute spectral cues or relative amplitude comparisons between frication and vowel for fricative identification, the frication and vowel were separated by (1) presenting the frication in isolation, and (2) inserting a gap of silence between the frication and vowel. The results showed that relative amplitude was perceived across vowel context and frication duration, and overrode context-dependent formant transition cues. The findings for temporal separations between the frication and vowel suggest that short-term memory processes may dominate the mediation of the relative-amplitude comparison. However, the overall results indicate that relative amplitude is only a component of spectral prominence, which is comprised of a primary frication spectral peak and a secondary frication/vowel peak comparison. PMID- 8227745 TI - Segment durations and accent as cues to word segmentation in Dutch. AB - This paper investigates the relative contributions of accentuation and of durational word boundary cues to listeners' perceived word segmentation in Dutch. A listening experiment is reported, in which 36 two-word phrases with an ambiguous word boundary were used as stimuli. Four groups of 20 subjects each had to make a forced binary choice between the two contrastive boundary positions. Accent position on either the first or second word was varied between stimulus phrases. In both contrastive realizations of each ambiguous phrase, two durational boundary cues were manipulated, viz. duration of the pivotal consonant and rise time of the postboundary vowel. Results show that durational word boundary markers are perceptually relevant for subjects' perceived word segmentation, but only if the manipulated segment is realized as word-initial. Second, accuracy in perceived word segmentation is higher if the second word (following the ambiguous boundary position) is accented. The latter result is probably due to the higher acoustic and perceptual salience of durational as well as spectral boundary cues, which occur in initial position in the second word. Hence, the contribution of accent to word segmentation may be partly due to its "enhancing" effect on phonetic word boundary markers. PMID- 8227746 TI - A comparison of speech discrimination with cochlear implants and tactile aids. AB - This study investigates patterns of speech discrimination in profoundly hearing impaired children who have received cochlear implants or tactile aids. The change/no change procedure was used to assess speech discrimination in these children. Three groups of subjects were tested: the first group used 3M/House single-channel cochlear implants; the second group used Nucleus 22-channel cochlear implants; and the third group used two-channel Tactaid II+ vibrotactile aids. Nine contrasts were constructed that assessed discrimination of suprasegmental and segmental speech features. Subjects were presented with stimulus trials in which stimuli changed during the trial or in which stimuli remained the same. Hits, misses, false alarms, and correct rejections were tallied and d' values were calculated for individual subjects for each contrast. Results indicated that different patterns of speech discrimination are provided by the three sensory prosthetic devices. For all contrasts, mean discrimination performance with the Nucleus device was better than that observed for the other two devices, despite the shorter duration of subject experience with this cochlear implant. In addition, interactions between device and speech contrast were not observed. Examination of individual subject performance revealed that each device group had a distribution of good to poor performers. The results suggest that the change/no change procedure is able to provide information regarding speech perception through sensory prosthetic devices despite existing differences in vocabulary and language skills of subjects. PMID- 8227747 TI - Simulation of the effects of loudness recruitment and threshold elevation on the intelligibility of speech in quiet and in a background of speech. AB - These experiments simulated the threshold elevation and loudness recruitment associated with three different types of cochlear hearing loss: Moderate flat (condition R2), severe flat (condition R3), and moderate-to-severe sloping (condition RX). This was done to allow an examination of the effects of these factors on the intelligibility of speech, in isolation from other factors that are normally associated with cochlear hearing loss, such as reduced frequency selectivity. The simulation was performed by splitting the input signal into 13 frequency bands, and processing the envelope in each band so as to create loudness sensations in a normal ear that would resemble those produced in an impaired ear with recruitment. The bands were then recombined. All tests were performed using subjects with normal hearing. For speech in quiet, simulation of hearing loss produced a reduction in the ability to understand low-level speech. However, speech at sufficiently high levels was highly intelligible in all conditions. Linear amplification according to the National Acoustic Laboratory (NAL) prescription gave high intelligibility for speech at normal conversational levels. For speech presented at a fixed input level of 65 dB SPL, against a background of a single competing talker, simulation of hearing loss produced substantial decrements in performance. The speech-to-background ratios in conditions R2 and RX had to be 11-13 dB higher than in the control condition (unprocessed stimuli) to achieve similar levels of performance. Linear amplification according to the NAL prescription improved performance markedly for the conditions simulating flat losses, but was less effective for the condition simulating a sloping loss. This indicates that threshold elevation combined with recruitment produces a loss of intelligibility for speech in the presence of a single competing talker that is only partly compensated by linear amplification of the type typically used in hearing aids. PMID- 8227748 TI - Cue trading in the production and perception of vowel stress. AB - The distinction between stressed and unstressed vowels in noun/verb pairs differentiated by stress (e.g., as in the word "misprint") is known to be cued by the acoustic attributes fundamental frequency, intensity, and duration. The role of these cues and vowel centralization measures in perception is ordered differently from that in production. It is not known what variability there is between speakers, whether the acoustic factors vary with speech clarity that a speaker produces on different occasions, or what the relationship is between the various speech attributes. Importantly, no studies have been performed in which production and perception of materials is compared with respect to these questions. In the current study, noun/verb pairs differentiated by stress were produced by four speakers, and measures of produced and perceived clarity obtained for these subjects. The importance of the acoustic factors to the contrast was modeled in perception and production, and the results compared. Additionally, the variation with produced or perceived clarity was ascertained. Finally, comparison between cue trading in production and perception was made. The results are discussed in connection with models of speech perception and production. PMID- 8227749 TI - The effect of temporal waveform shape on spectral discrimination by normal hearing and hearing-impaired listeners. AB - A listener's ability to discriminate differences in amplitude spectra is generally thought to be independent of the temporal waveform shapes of the stimuli. However, there have been reports of enhanced contrast between spectral peaks and valleys of harmonic complexes represented in the temporal firing patterns of auditory-nerve fibers for some combinations of component amplitudes and phases [e.g., Horst et al., J. Acoust. Soc. Am. 88, 2656-2681 (1990)]. The enhancement has been attributed to nonlinear processing of high-amplitude peaked waveforms. To determine whether such enhancements are observed in psychoacoustic responses of human listeners, subjects were asked to discriminate between harmonic complexes that differed in the frequency location of three formantlike increments in harmonic amplitude. The stimuli were constructed to have either a very flat or a very peaked temporal envelope. Over a range of 1- to 10-dB spectral contrast between elevated harmonics and background harmonics, discrimination ability was determined as a function of waveform shape and overall amplitude. Performance was measured for both normal-hearing and hearing-impaired listeners. Normal-hearing listeners showed lower thresholds of spectral contrast for peaked waveforms at high intensities than for any of the other phase amplitude conditions. Although the effect was small (1 to 2 dB), it is consistent with an hypothesized enhancement due to nonlinear cochlear processing. Hearing impaired listeners did not demonstrate differential performance across phase or amplitude conditions, perhaps reflecting more linear processing in these damaged ears. PMID- 8227750 TI - Auditory spectral discrimination and the localization of clicks in the sagittal plane. AB - Experiments show that the ability of human listeners to localize an impulsive sound in the medial sagittal plane (front, overhead, rear) deteriorates as the level of the sound increases. This negative level effect is strong for clicks but does not appear for broadband noise. It is conjectured that the negative level effect arises because the tonotopic excitation pattern is broadened for intense impulsive sounds. As a result, the spectral peaks and valleys, which are caused by anatomical filtering and which normally code for localization in the sagittal plane, are less recognizable. Filtered click discrimination experiments using headphones also show a negative level effect for clicks, but not for noise, and support this conjecture. PMID- 8227751 TI - Vulnerability and adaptation of distortion product otoacoustic emissions to endocochlear potential variation. AB - The endocochlear potential (EP) was reversibly decreased in adult gerbils by the intraperitoneal injection of furosemide, while cochlear functioning was monitored by measurement of distortion production otoacoustic emissions (DPE) at a range of stimulus intensities. Stimulus frequencies for DPEs were f1 = 6.8 and f2 = 8 kHz (f2/f1 = 1.18). Emissions monitored in the ear canal and scala media were 2f1-f2, 3f1-2f2, 2f2-f1, and f2-f1. Typically, the EP decreased smoothly, reached a minimum one-half hour after injection, then recovered slowly over several hours. Emissions at 2f1-f2 and 3f1-2f2 at low stimulus levels were particularly vulnerable to the change in EP. These vulnerable emissions showed characteristic trajectories in which the amplitudes changed little with the initial EP decrease, then dropped sharply as the EP continued to decrease. However, the amplitudes then began to recover even before the EP reached minimum, and recovered completely while the EP remained subnormal. The trajectories of the other odd order emissions were similar, but lacked the abrupt decrease. The variation of the even order (f2-f1) component was completely different, but appeared related to the odd order trajectories in a complex fashion. During the initial decrease for the vulnerable components, the decrease in emission amplitude (in dB) was found to be proportional to the square of the change in EP (in mV). The recovery with a subnormal EP was interpreted as an adaptive effect with a time constant of about 15 min. PMID- 8227752 TI - Functional changes in the ventral cochlear nucleus following acute acoustic overstimulation. AB - The effects of acute acoustic overstimulation on the discharge patterns of neurons in the ventral cochlear nucleus (VCN) were evaluated in anesthetized chinchillas. Response measures were obtained from the same neuron before and after presenting a 3- to 5-min intense tone (90-105 dB SPL) located one-half oct above the unit's characteristic frequency (CF). If a unit had an inhibitory response area at frequencies above CF and if the traumatizing tone reduced the magnitude of the inhibitory response, then the neuron's discharge rate to suprathreshold tones at CF increased ("enhancement") by as much as 25%. However, if a unit lacked an inhibitory response area at frequencies above CF, then the traumatizing tone typically caused either no change or a decrease in the unit's discharge rate at CF. The traumatizing tone did not alter the shape of the post stimulus time histograms. Moreover, the width of the excitatory response area was not altered by the exposure even when traumatizing stimulus reduced the magnitude of the inhibitory response above CF. The enhanced firing rate at CF following the exposure could conceivably contribute to the enhanced evoked-potential amplitudes observed in the auditory brain stem following acoustic trauma. PMID- 8227753 TI - Gender differences in cochlear response time: an explanation for gender amplitude differences in the unmasked auditory brain-stem response. AB - Derived narrow-band auditory brain-stem responses (ABRs) in young normal-hearing subjects revealed a significant gender difference in response time between frequency regions of the cochlea. Females showed shorter delays than males between derived bands. This differential has not been previously reported. As in many early studies, the unmasked amplitude of the wave V complex was significantly larger (30%) in females than males. However, differences in amplitudes of the narrow-band responses were too small to account for the differential in the unmasked response. It is hypothesized that the larger amplitude of the unmasked wave V complex in females occurs because of a faster response time across the cochlea leading to better neural synchrony and, therefore, larger amplitudes. Furthermore, results can be explained by assuming that the stiffness gradient in the cochlea is 13% larger in females than in males. If males and females have the same cochlear tonotopic mapping, the female cochlea should be 13% shorter. This prediction is highly consistent with recent anatomical studies of cochlear length and gender. The results of the present study indicated possibly important cochlear mechanisms that influence the main parameters of ABRs. An understanding of these cochlear mechanisms may improve the diagnostic capabilities of ABRs in patients with peripheral hearing loss. PMID- 8227754 TI - The relation between head size and auditory brain-stem response interpeak latency maturation. AB - In developmental populations, duration of auditory brain-stem response (ABR) I III, III-V, and I-V vary substantially across individuals, particularly among preterm infants. Adult ABR interpeak latency has a strong correlation with brain stem size and weaker correlation with head size. To determine if head size might contribute to this increased interpeak latency variability among infants, ABR data were normalized based on head circumference. Normalization by head circumference did not reduce interpeak variability. Further analyses revealed a negative correlation between interpeak latency and head circumference that varied as a function of age. Before 42 weeks conceptional age (CA), a significant relation exists between increased head circumference and decreased duration of the III-V and I-V intervals, but not the I-III interval. For infants older than 42 weeks CA, there was a significant relation between increased head circumference and decreased duration for the I-III intervals but not the III-V and I-V intervals. An age-dependent correlation between decreasing interpeak latency and increasing head circumference suggests that improved neural transmission through the auditory nerve and brain-stem pathway offset or even overcompensate for developmental lengthening of the sensory pathway. Also, developmental time constants obtained from nonlinear curve fit analyses were shorter for normalized than non-normalized data, particularly for the I-V interval. Therefore, correction of ABR data for the length of the sensory pathway may be important to estimate accurately maturation rate for developmental populations. PMID- 8227755 TI - The relation among fundamental frequency, intensity, and duration varies with accentuation. PMID- 8227756 TI - Some acoustic effects of listening to noise on speech production. PMID- 8227757 TI - Comments on "Stimulus dependencies of the gerbil brain-stem auditory-evoked response (BAER). I: Effects of click level, rate and polarity" [J. Acoust. Soc. Am. 85, 2514-2525 (1989)]. PMID- 8227758 TI - The comparative efficacy of carbamazepine low and high serum level and lithium carbonate in the prophylaxis of affective disorders. AB - The prophylactic efficacy of carbamazepine slow release (CBZ) at ke different blood levels and lithium carbonate slow release (LI) was compared in a retrospective/prospective, randomized, 2-year open trial. 84 patients with a DSM III-R diagnosis of recurrent affective disorder who had no prophylactic medication in the 2 years preceding the trial (no LI nonresponders), were randomly allocated to three treatment groups: CBZ low (15-25 mumol/l), CBZ high (28-40 mumol/l) and LI (0.6-0.8 mumol/l). Fifty-eight patients completed the full observation period of 2 years, 26 patients dropped out. There were no statistically significant differences in the efficacy of the prophylactic treatment for bipolar patients. For the unipolar patients, the group with a low CBZ serum level showed no reduction in the duration of episodes. The two other treatment groups seem to be equal in attenuation of a unipolar course of an affective disorder. PMID- 8227759 TI - 99mTc-HMPAO single photon emission tomography in late life depression: a pilot study of regional cerebral blood flow at rest and during a verbal fluency task. AB - Relative regional cerebral blood flow (rCBF) was measured at rest and during a verbal fluency task in ten elderly patients with major depressive disorder and nine controls. At rest, depressed patients showed significant reductions in the cortico-cerebellar ratios of tracer uptake in the right and left parietal, left temporal and left occipital regions. During the task relative rCBF increased in patients such that these differences disappeared. Relative rCBF did not correlate with severity of depressed mood or endogenous features but did correlate positively with severity of psychotic symptoms. Frontal relative rCBF correlated negatively with somatic symptoms and anxiety. PMID- 8227760 TI - Can disulfiram potentiate the dexamethasone suppression test in depressive patients? AB - Transient DST non-suppression during disulfiram-treatment was fortuitously observed in a depressed patient. A disulfiram-potentiated DST (DSTd) was constructed by giving 800 mg disulfiram on the dexamethasone and postdexamethasone days. DST and DSTd were compared intraindividually in 21 patients and 14 controls. Disulfiram-potentiation was significant in constantly depressed patients, and in questionable controls due to possible subclinical depression or somatic factors which may cause 'false-positive' results. It did not occur in depressed patients during marked improvement or long-term recovery, and not in accurately screened controls. DSTd was significantly correlated with depression ratings, whereas DST was not. PMID- 8227761 TI - Psychiatric impact of the loss of an adolescent sibling to suicide. AB - Twenty-five adolescent siblings of 20 adolescent suicide victims were psychiatrically assessed 6 months after the suicide, and compared to 25 demographically matched controls. Siblings were much more likely to show a new onset major depression subsequent to exposure to suicide. New-onset depression was associated with previous psychiatric disorder, family history of any psychiatric disorder, and family history of major depression. Mothers of suicide victims, compared to the mothers of controls, were also more likely to be depressed 6 months after the suicide of their child. The development of a family based intervention for families of adolescent suicides is recommended. PMID- 8227762 TI - The recruitment process for a multicenter study on the long-term prophylactic treatment of affective disorders. AB - The paper reports on the process of patient recruitment for a controlled clinical multicenter study on the treatment of affective disorders. Two thirds of the patients screened did not participate because prophylactic treatment was either unnecessary or not justified for medical reasons. Further, a number of patients equal to that eventually allocated to the trial refused to participate for personal, idiosyncratic reasons. In spite of this, the patients in the trial were very similar to those not participating with respect to relevant variables such as age, sex, number of and intervals between previous episodes or severity of the present episode. PMID- 8227763 TI - Expressive characteristics of anxiety in depressed men and women. AB - This study was aimed at identifying the expressive, movement, and social behaviors associated with anxiety in the syndrome of major depression. The sample consisted of 97 hospitalized male and female depressed patients. Expressive and social behaviors were evaluated prior to treatment in a structured videotaped interview. Anxiety was measured using a multi-vantaged approach including doctor's rating, nurse's rating, patient self-report, and a separate video rating. Results indicate that anxiety was significantly associated with agitation, distressed facial expression, bodily discomfort, and poor social interaction in both sexes. Men and women differed in certain respects: anxiety was highly related to motor retardation in women only, and to hostility in men only. Differences in the pattern of expressive behavior between high and low anxious, depressed patients were clearly significant, and several were large enough to serve as clinical indicators. These findings help to characterize the expressive features of anxiety in the context of severe depression, and add to the growing literature on sex differences in depression. PMID- 8227764 TI - Psychotic symptoms in depression and borderline personality disorder. AB - The authors investigated the prevalence of psychotic symptoms in depression and borderline personality disorder employing the Diagnostic Interview for Borderlines. Clear-cut delusions and hallucinations were rare among the borderlines. However, derealization and depersonalization symptoms were common and were found to be prevalent as among depressives. The prevalence of these symptoms among patients with both borderline personality disorder and depression was similar to that among patients with only borderline personality disorder or depression. The relationship between depression and borderline personality disorder and the significance of psychotic symptoms in these disorders is discussed. PMID- 8227765 TI - Dexamethasone suppression test and TRH-TSH test in subaffective dysthymia and character-spectrum disorder. AB - Based on Akiskal's criteria of subaffective dysthymia (SDT) and character spectrum disorder (CSD) as the two, etiologically distinct forms of early-onset primary dysthymia, the authors investigated the dexamethasone suppression test (DST) in 18 patients with SDT and in 30 patients with CSD. TRH-TSH test was also investigated in smaller subsamples of the patients (n = 8, and n = 7, respectively). Fifty percent of the patients with SDT showed abnormal DST and TRH TSH test results respectively, while the figures in the CSD patients were 7% and 0%. These findings suggest that SDT is a clinically diagnosable and biologically distinct subgroup within the broader category of early-onset primary dysthymia, which represents a symptomatically milder version of primary affective disorder. PMID- 8227766 TI - An issue of dental necessity. PMID- 8227767 TI - Dental office emergencies. PMID- 8227768 TI - Legal obligations. PMID- 8227769 TI - One house. PMID- 8227770 TI - One degree. PMID- 8227771 TI - Thoughts on TMD. PMID- 8227772 TI - Evaluation and control of post-cementation pulpal sensitivity: zinc phosphate and glass ionomer luting cements. AB - Many studies have documented pulpal sensitivity after crown cementation, but none have determined its cause. By controlling technique variables in a large-scale clinical trial, the authors evaluated the contribution of zinc phosphate and glass ionomer luting cements in causing pulpal sensitivity or necrosis. PMID- 8227773 TI - Systemic osteoporosis and oral bone loss: evidence shows increased risk factors. AB - Aging is associated with a loss of bone mass and an increased risk of oral and systemic bone loss. Systemic osteoporosis can result in bone fractures, especially of the spine and hip, with the characteristic spinal curvature and loss of height often seen in osteoporotic postmenopausal women. Loss of teeth and ridge resorption can occur in the mouth. PMID- 8227774 TI - Using triazolam to reduce dental anxiety. AB - Triazolam, commonly prescribed to treat insomnia, also can be used to reduce dental anxiety. Despite controversial reports in the lay press, triazolam can be used safely at low doses for short periods. The authors review research concerning the drug's safety and discuss its use in dentistry. PMID- 8227775 TI - Clinical evaluation of a universal dentin bonding resin: preserving dentition through new materials. AB - The new generation of dentin bonding materials can withstand the contraction shrinkage of composite materials. A two-year clinical trial of one material showed an excellent retention rate. PMID- 8227776 TI - Treating Kaposi's lesions in the HIV-infected patient. AB - Kaposi's sarcoma is the most frequent malignant neoplasm in AIDS, occurring in about 10 percent of all risk groups. This study evaluates the effect of intralesional vinblastine on intraoral Kaposi's sarcoma in 24 HIV-positive, homosexual males with 82 lesions. Complete resolution occurred in nearly 70 percent of the cases. PMID- 8227777 TI - Dental fees: a candid discussion. AB - Many dental fees have been determined illogically since the beginning of dentistry. Setting fees, however, can be relatively easy and quite equitable for all concerned if they are based on clinical time involved, procedure difficulty and quality of service. Our goal should be to provide the highest quality oral care possible at the lowest fees. It can be done. But changes need to be made, individually and as a profession, in fee structures and the manner in which we deliver treatment. PMID- 8227778 TI - The esthetic impact of orthodontics: planning treatment to meet patients' needs. AB - Successful orthodontic therapy in the growing patient involves the coordination of mechanotherapy, growth modification techniques, knowledge of hard and soft tissue growth patterns and patient motivational techniques. These cases represent differing types of Class II malocclusions in which the causes and treatment solutions were quite varied. The differing esthetic needs used treatment plans designed after a careful, problem-oriented assessment of the patient's needs. In each case, the success involved appropriate timing, differential diagnosis, excellent patient cooperation and the design of treatment plans to satisfy occlusal and esthetic goals. Teamwork between general practitioners and orthodontists is critical in meeting patients' needs. PMID- 8227779 TI - A multicenter randomized trial comparing a percutaneous collagen hemostasis device with conventional manual compression after diagnostic angiography and angioplasty. AB - OBJECTIVES: A new percutaneous collagen hemostasis device was compared with conventional compression techniques after diagnostic catheterization and angioplasty. BACKGROUND: Peripheral vascular complications after diagnostic catheterization or more complex interventional procedures, as well as the discomfort of manual compression and prolonged bed rest, represent significant morbidity for invasive cardiac procedures. METHODS: A prospective, multicenter, randomized trial was designed to compare the hemostasis time in minutes and the incidence of vascular complications in patients receiving a vascular hemostasis device with those undergoing conventional compression techniques. RESULTS: After diagnostic catheterization, hemostasis time was significantly less with the vascular hemostasis device than with conventional manual compression (4.1 +/- 2.8 min [n = 90 patients] vs. 17.6 +/- 9.2 min [n = 75], p < 0.0001). This difference was greater in patients undergoing angioplasty and was unrelated to the anticoagulation status (4.3 +/- 3.7 min [n = 71 not receiving heparin], 7.6 +/- 11.6 min [n = 85 receiving heparin], 33.6 +/- 24.2 min [n = 134 control patients not receiving heparin], p < 0.0001 vs. control patients). The time from the start of the procedure to ambulation was slightly less after diagnostic catheterization in patients treated with the device (13.3 +/- 12.1 h vs. 19.2 +/- 17.8 h, p < 0.05). It was also less in patients who underwent angioplasty when the device was used after discontinuation of anticoagulation (23.0 +/- 11.1 h, without heparin), as compared with control compression techniques (32.7 +/- 18.8 h, p < 0.0001). Time to ambulation was even shorter (16.1 +/- 11.1 h, p < 0.0001) in patients in whom the device was placed immediately after angioplasty while they were still fully anticoagulated with a prolonged activated clotting time (336 +/- 85 s). There were no major complications (surgery or transfusion) after diagnostic catheterization and a low incidence of major complications in patients who underwent angioplasty (0.7% in control patients, 1.4% with the device without heparin, 1.2% with the device and heparin, p = NS). After angioplasty, there was a trend toward fewer hematomas when the device was used in the absence of heparin (4.2% vs. 9.7% in control patients, p = 0.14). CONCLUSIONS: A new vascular hemostasis device can significantly reduce the puncture site hemostasis time and the time to ambulation without significantly increasing the risk of peripheral vascular complications. The role of this technology in reducing complications, length of hospital stay and cost remains to be determined. PMID- 8227780 TI - Utility of a percutaneous collagen hemostasis device: to plug or not to plug? PMID- 8227781 TI - Coronary embolization after balloon angioplasty or thrombolytic therapy: an autopsy study of 32 cases. AB - OBJECTIVES: This study was undertaken to examine the nature, extent and clinical relevance of coronary embolism after balloon angioplasty or thrombolytic therapy, or both. BACKGROUND: Histopathologic documentation of postinterventional coronary embolization has been reported in only 10 patients from five studies. METHODS: This retrospective autopsy-based study included 32 patients, treated with balloon angioplasty or thrombolysis, or both, who died within 3 weeks of the procedure and underwent autopsy at the Mayo Clinic. Clinical variables included patient age and gender, artery treated, site and type of obstruction, type of intervention, success of the procedure, and postprocedural changes in the electrocardiogram (ECG), cardiac enzymes and hemodynamic status. Histopathologic variables included characteristics of treated plaques, acutely infarcted myocardium and coronary microemboli. Associations between microemboli and clinical and microscopic factors were evaluated by t tests and simple and multiple linear regression. RESULTS: Emboli were observed in 26 (81%) of the 32 patients. Among 83 emboli, 95% were thrombotic or atheromatous. The presence of microemboli was associated statistically with the development of postprocedural infarct extension, new myocardial infarction or new ECG abnormalities. Moreover, the greatest number of microemboli were associated with intervention in the left anterior descending coronary artery, multiple interventional sites, postprocedural medial dissection and plaque rupture or extrusion. CONCLUSIONS: Among patients undergoing balloon angioplasty or thrombolytic therapy who die and undergo autopsy, coronary microemboli occur in a substantial percent. The frequency in survivors is unknown. However, in living patients who develop acute myocardial ischemia or new ECG abnormalities after these interventions, coronary microembolization should be considered a potential cause. PMID- 8227782 TI - Is traditionally defined complete revascularization needed for patients with multivessel disease treated by elective coronary angioplasty? Multivessel Angioplasty Prognosis Study (MAPS) Group. AB - OBJECTIVES: The purpose of this study was to determine the effect of incomplete revascularization by percutaneous transluminal coronary angioplasty in patients with multivessel disease on adverse long-term cardiac events (death, coronary artery bypass surgery or myocardial infarction) and to develop an optimal definition of adequate revascularization based on clinical outcome. BACKGROUND: The effect of incomplete coronary revascularization by coronary angioplasty on long-term adverse clinical events remains controversial. METHODS: Three hundred seventy well characterized patients were followed-up for 27 +/- 16 months after angioplasty. Mean patient age was 58 +/- 11 years; 72% were male; 70% had two vessel disease (> or = 50% diameter stenosis by caliper measurement); and the mean left ventricular ejection fraction was 58 +/- 11% (range 20% to 85%). Angioplasty was successfully accomplished in 339 patients (91.6%), but complete revascularization by the standard definition (no residual > or = 50% stenosis in a coronary artery > or = 1.5 mm in diameter) was achieved in only 91 patients (25%). RESULTS: Three-year event-free survival (i.e., freedom from death, myocardial infarction, coronary artery bypass surgery) in the entire cohort was 76.5%. By the standard definition, complete revascularization was strongly and negatively associated (p = 0.003) with long-term cardiac events, even after correction for the effects of other independent correlates of events, using Cox proportional hazard regression analysis. Seventeen other definitions, evaluating the severity and extent of residual stenoses and whether they were associated with contractile myocardium, were tested to find that which best stratified late event-free survival and had an outcome with complete revascularization no worse than that associated with the standard definition. The best definition for the entire cohort, having more predictive value than the standard definition, allowed < 10% of estimated left ventricular mass to be served by vessels with mild stenoses (< 60%) without being considered "incomplete." CONCLUSIONS: Mild stenoses in coronary arteries > or = 1.5 mm in diameter serving modest amounts of myocardium do not appear to need to be revascularized to achieve good long-term outcome with coronary angioplasty. Hence, angioplasty in such lesions may not be justified except when they are documented to cause life-style-limiting angina, and the standard definition of complete revascularization by angioplasty appears to be suboptimal. The importance of optimally defined adequate revascularization should be considered in the interpretation of the results of randomized trials assessing the clinical efficacy of coronary angioplasty compared with that of other modalities of therapy. PMID- 8227783 TI - Percutaneous transluminal coronary angioplasty of one major coronary artery when the contralateral vessel is occluded. AB - OBJECTIVES: In 193 patients we evaluated the safety and efficacy of angioplasty of a critical stenosis of the right coronary artery (52 patients) or the left anterior descending coronary artery (141 patients), with the contralateral coronary artery occluded and the circumflex artery being without significant stenosis. BACKGROUND: Attempted angioplasty of either the left anterior descending or the dominant right coronary artery when the contralateral vessel is occluded may trigger overwhelming left ventricular dysfunction or hemodynamic collapse, or both. METHODS: Immediate and late outcome (33 +/- 18 months) in the study group were compared with outcome in 214 patients who had angioplasty in both the left anterior descending and right coronary arteries and in 194 patients who had coronary artery surgery and were matched for number and location of significant lesions, ejection fraction, age, gender and study period. RESULTS: Left ventricular function was normal (38%) or mildly (34%), moderately (22%) or severely (6%) compromised. There were 11 (5.7%) emergency and 5 (2.6%) elective coronary artery operations, 3 (1.6%) myocardial infarctions and 1 in-hospital death in the study group. After discharge there were 25 (13.1%) elective coronary operations, 7 (3.7%) myocardial infarctions and 9 (4.7%) deaths in the study group. The incidence of death and myocardial infarction was similar in all groups, with 80% power to detect a 7% difference in adverse events. The study group had more elective surgery before and after discharge than did the surgical control group (p = 0.02). CONCLUSIONS: Dilating one major vessel when the contralateral vessel is occluded appears to be as safe as coronary surgery or two vessel angioplasty. Incomplete revascularization in study group patients did not impair survival or increase myocardial infarction compared with the angioplasty and surgical control groups. PMID- 8227784 TI - Prehospital thrombolysis: beneficial effects of very early treatment on infarct size and left ventricular function. AB - OBJECTIVES: The purpose of this study was to compare the effects of very early (< or = 1.5 h after symptom onset) and later (> 1.5 up to 4 h) thrombolytic therapy on infarct size, left ventricular function and early mortality in patients with acute myocardial infarction. To start thrombolysis at the earliest possible moment, it was performed in the prehospital setting. A cutoff time of 1.5 h was prospectively stipulated. BACKGROUND: Shortening of ischemic time is crucial within the 1st 2 h. Prehospital thrombolysis can reduce time to treatment and enables very early initiation of therapy for many patients. METHODS: One hundred seventy patients received 30 mg of anistreplase up to 4 h from symptom onset by a mobile intensive care unit physician. Infarct size was measured from cumulative release of alpha-hydroxybutyrate dehydrogenase, and left ventricular function was assessed by contrast angiograms 10 days after the infarction. RESULTS: The decision to treat on scene was correct in 98% of patients. There were no bleeding complications or deaths outside the hospital setting. In 28 patients (17%) the ischemic process was interrupted. Findings with thrombolytic therapy initiated < or = 1.5 (96 patients) versus > 1.5 h (74 patients) were the following: initial extent of epicardial injury, 1.6 +/- 0.9 versus 1.4 +/- 0.7 mV, p = NS; infarct size by cardiac enzyme release 646 +/- 634 versus 886 +/- 712 IU/liter, p < 0.05; ejection fraction 57 +/- 14% versus 51 +/- 13%, p < 0.05; regional dyssynergic area 24 +/- 22 versus 33 +/- 24 U, p < 0.05; 21-day mortality 1 of 96 versus 5 of 74 patients (1% vs. 7%, p < 0.05). CONCLUSIONS: The data suggest that in evolving myocardial infarction up to 4 h in duration, the start of thrombolytic therapy at < or = 1.5 h compared with > 1.5 h limits infarct size, preserves left ventricular function and may save lives. PMID- 8227785 TI - Severity and response of chest pain during thrombolytic therapy for acute myocardial infarction: a useful indicator of myocardial salvage and infarct size. AB - OBJECTIVES: The purpose of this study was to determine noninvasively whether chest pain severity is predictive of the amount of myocardium at risk and whether the response of pain during thrombolysis is associated with myocardial salvage during acute myocardial infarction. BACKGROUND: The perception of chest pain and response to reperfusion therapy during acute myocardial infarction may provide important information for treatment benefit. Previous studies have been limited by the inability to measure myocardium at risk and myocardial salvage. METHODS: Sixty-two patients with acute myocardial infarction received an injection of technetium-99m sestamibi before thrombolysis and again at hospital discharge. Tomographic imaging was performed 1 to 6 h later. Myocardium at risk, infarct size and absolute myocardial salvage were derived from these images using previously described techniques and were expressed as a percent of the left ventricle. Salvage index was calculated by dividing myocardial salvage by the myocardium at risk. Chest pain severity was graded before thrombolysis as none, mild, moderate or severe. Chest pain response during thrombolytic therapy was graded as none, partial or completely resolved. RESULTS: There was no association between chest pain severity and myocardium at risk, but there was a weak trend toward greater myocardial salvage and salvage index (p = 0.09 and p = 0.12, respectively) for patients with more severe symptoms. Patients without chest pain at the start of thrombolysis still demonstrated significant salvage (11 +/- 11% of the left ventricle, p = 0.009). There was a significant association between chest pain response to therapy and both myocardial salvage (p = 0.03) and salvage index (p = 0.01). By multivariate analysis, chest pain severity and response of chest pain during thrombolysis were significant independent predictors of myocardial salvage, salvage index and infarct size. Thrombolysis was most effective in the 20 patients (32%) with moderate or severe chest pain and complete resolution of symptoms during thrombolysis (salvage of 79% to 89% of the area at risk). In the remaining 32 patients with chest pain, salvage of the area at risk was only 32%. CONCLUSIONS: These findings suggest that the assessment of chest pain before and after thrombolytic therapy is a readily available, useful indicator of the efficacy of the therapy. PMID- 8227786 TI - Bedside cardiology and thrombolysis. AB - The study by Christian et al. (37) provides useful clinical information for the management of the patient with myocardial infarction with thrombolytic therapy. 1) Complete resolution of chest pain during the infusion of an intravenous thrombolytic agent suggests reperfusion and myocardial salvage. Initial conservative medical management should be considered in these patients especially if serial ECGs show a progressive and rapid downward defection of the ST segment. 2) Patients presenting within 6 h after the onset of infarction, who are pain free, may still benefit from thrombolysis if their ECGs show persistent ischemia. 3) In the remaining patients in whom chest pain does not resolve completely during thrombolytic therapy, management, whether continued medical or invasive strategy, should be individualized and dictated by the extent of myocardium at risk (i.e., by the number of ECG leads showing ST segment elevation), the response of the ST segment to thrombolysis and, most important, the clinical status of the patient. PMID- 8227787 TI - Effects of low dose transdermal scopolamine on heart rate variability in acute myocardial infarction. AB - OBJECTIVES: We hypothesized that by enhancing parasympathetic activity, low dose transdermal scopolamine would increase heart rate variability after myocardial infarction. BACKGROUND: Low heart rate variability is associated with increased mortality after acute myocardial infarction. METHODS: Conventional time domain heart rate variability was measured from 24-h Holter recordings of 61 consecutive male patients (mean age 58 +/- 10 years, left ventricular ejection fraction 44.7 +/- 15.5%) 6 days (median) after acute myocardial infarction. Patients were then randomly assigned to wear one patch of transdermal scopolamine or a matching placebo patch for 24 h, during which their 24-h heart rate variability was remeasured. RESULTS: Compared with placebo, transdermal scopolamine caused a significant increase in time domain measures of 24-h heart rate variability by 26% to 35% above baseline. Transdermal scopolamine was well tolerated. CONCLUSIONS: Low dose transdermal scopolamine safely increases cardiac parasympathetic activity and short-term heart rate variability after acute myocardial infarction. Whether the effect of transdermal scopolamine on heart rate variability is a reasonable surrogate for improvement of long-term morbidity and mortality requires an appropriate designed investigation. PMID- 8227788 TI - Scopolamine increases vagal tone and vagal reflexes in patients after myocardial infarction. AB - OBJECTIVES: The goal of this study was to assess the hypothesis that transdermal scopolamine would increase vagal activity in patients after myocardial infarction. BACKGROUND: In postmyocardial infarction patients, low heart rate variability and reduced baroreceptor reflex sensitivity are associated with increased mortality. Accordingly, there is an increasing interest in a mechanism for shifting the sympathovagal balance toward vagal dominance. METHODS: The effects of transdermal administration of scopolamine on heart rate variability and baroreceptor reflex sensitivity were assessed in 20 patients (mean age 59 +/- 11 years) by pharmacologic washout 14 +/- 3 days after myocardial infarction. Heart rate variability and baroreceptor reflex sensitivity were measured 24 h after application of the scopolamine patch and compared with the values measured before scopolamine and after application of a placebo patch. The following variables were derived from a 15-min electrocardiographic recording: the mean RR interval and its standard deviation, the mean square successive difference, the percent of intervals differing > 50 ms from the preceding RR interval and the low and high frequency areas resulting from power spectral analysis. RESULTS: The placebo patch had no effect on the variables measured. Scopolamine increased both heart rate variability and baroreceptor reflex sensitivity significantly. Specifically, the mean RR interval and its standard deviation increased by 7.1% (p = 0.01) and 25% (p = 0.004), respectively. The mean square successive difference increased by 38% (p = 0.0003) and the percent of intervals differing > 50 ms from the preceding interval by 100% (p = 0.001). The ratio of low to high frequency areas of the power spectrum decreased by 24% (p = 0.02), and baroreceptor reflex sensitivity increased by 42% (p = 0.0006). These effects were also evident in patients with very low initial values. Side effects were minimal. CONCLUSIONS: Transdermal scopolamine increased measures of heart rate variability and baroreceptor reflex sensitivity in patients with a recent myocardial infarction toward values associated with a better prognosis. Pharmacologic modulation of the autonomic balance by scopolamine or related drugs deserves evaluation as a new and promising approach to reduce risk after myocardial infarction. PMID- 8227789 TI - Vagal stimulation after myocardial infarction: accentuating the positive. PMID- 8227790 TI - Regional cardiac sympathetic denervation in patients with ventricular tachycardia in the absence of coronary artery disease. AB - OBJECTIVES: The aim of this study was to determine whether patients with ventricular arrhythmias in the absence of coronary artery disease also have abnormalities in sympathetic innervation. BACKGROUND: We have previously shown by cardiac sympathetic scintigraphy using iodine-123-metaiodobenzylguanidine (I-123 MIBG) that patients with ventricular tachycardia after myocardial infarction have regional cardiac sympathetic denervation. It is not known whether patients with ventricular tachycardia in the absence of coronary artery disease also have regional cardiac sympathetic denervation. METHODS: We performed cardiac I-123 MIBG and thallium-201 single-photon emission computed tomographic (SPECT) scans at rest in 18 patients (mean age 47 +/- 18 years) with cardiomyopathy (n = 6), left ventricular hypertrophy (n = 1), valvular disease (n = 2) or a structurally normal heart (n = 9) who presented with monomorphic (n = 15) or polymorphic (n = 3) ventricular tachycardia. These scans were compared with scans in 12 control patients without ventricular tachycardia (mean age 30 +/- 17 years) who had cardiomyopathy (n = 3) or a structurally normal heart (n = 9). Cardiac sympathetic denervation was defined as myocardial areas having thallium uptake with reduced or absent I-123-MIBG uptake. RESULTS: Twelve (67%) of 18 patients with ventricular tachycardia had regional cardiac sympathetic denervation compared with 1 (8%) of 12 patients who did not have ventricular tachycardia (p = 0.002). In the nine patients with a structurally normal heart and ventricular tachycardia, five (55%) patients had regional cardiac sympathetic denervation compared with zero of nine control patients with a structurally normal heart (p = 0.029). Five patients underwent right ventricular radiofrequency ablation for ventricular tachycardia, and sympathetic denervation was adjacent to the ablation site in one of these patients. CONCLUSIONS: Patients with ventricular tachycardia in the absence of coronary artery disease have abnormal cardiac sympathetic innervation detectable by cardiac sympathetic scintigraphy. The role of regional cardiac sympathetic denervation in arrhythmogenesis remains to be determined. PMID- 8227791 TI - External cardiac pacing using low impedance electrodes suitable for defibrillation: a comparative blinded study. AB - OBJECTIVES: The objective of this study was to determine whether the threshold for successful cardiac pacing is affected by electrode impedance and whether this procedure can be successfully carried out through low impedance electrodes that are also suitable for defibrillation. BACKGROUND: Reintroduced in the early 1980s, external cardiac pacing utilizes large externally placed electrodes with a high impedance, in conjunction with a stimulator capable of producing an impulse of 20 to 40 ms in duration. On the basis of empiric observation, high impedance electrodes (> 500 omega) are believed to be optimal for external cardiac pacing. Such electrodes are unsuitable for defibrillation, a technique that is most successful when impedance is low. In view of the absence of controlled data to support this recommendation, as well as the desirability of using one set of electrodes for both pacing and defibrillation, we undertook the following study. METHODS: Thirty-two normal subjects underwent a total of 110 attempts at external cardiac pacing with either (or both) high or low impedance electrodes in combination with one or two commercially available external cardiac pacemakers. Each subject underwent pacing at least twice in a randomized double-blind fashion to determine the pacing threshold and level of discomfort. RESULTS: Individual subjects had a wide range of pacing thresholds but did not experience any greater discomfort with one pacemaker-electrode combination than with any other. Similarly, no pacemaker-electrode combination was superior to another in terms of pacing thresholds. The mean pacing threshold was 72.5 +/- 6 mA for the 40-ms impulse/high impedance electrode combination, 78.7 +/- 6 mA for the 40-ms impulse/low impedance electrode, 73.8 +/- 7 mA for the 20-ms impulse/high impedance electrode and 77.5 +/- 7 mA for the 20-ms impulse/low impedance electrode (p = NS for all comparisons). CONCLUSIONS: Contrary to previous belief, a high impedance electrode offers no advantage for external pacing in terms of either pacing threshold or discomfort level during pacing. This study demonstrates that it is feasible to perform external pacing through an electrode that is also suitable for defibrillation and suggests that a single external pacing-defibrillation electrode is all that is needed to perform these two procedures. PMID- 8227792 TI - Impact of electrical cardioversion for atrial fibrillation on left atrial appendage function and spontaneous echo contrast: characterization by simultaneous transesophageal echocardiography. AB - OBJECTIVES: This study assessed the function of the left atrial appendage in the pericardioversion period to gain insights into mechanisms involved in thromboembolism after cardioversion of atrial fibrillation. BACKGROUND: Systemic embolization associated with electrical cardioversion of atrial fibrillation is thought to originate from the left atrium or left atrial appendage, or both. However, the mechanism involved is poorly understood. METHODS: We studied left atrial appendage function with transesophageal echocardiography in 20 patients with atrial fibrillation before and after successful electrical cardioversion. We measured left atrial appendage emptying and filling velocities by pulsed wave Doppler echocardiography, characterized Doppler emptying patterns, measured atrial appendage areas and assessed the presence or absence of spontaneous echo contrast or thrombus. RESULTS: Organized left atrial appendage function returned in 16 (80%) of 20 patients immediately after cardioversion. Atrial appendage emptying velocities before cardioversion were greater in patients without (0.39 +/- 0.02 m/s) than in those with (0.25 +/- 0.12 m/s) spontaneous echo contrast (p = 0.045). Furthermore, emptying velocities before cardioversion were significantly greater than late diastolic emptying velocities after cardioversion (0.31 +/- 0.15 vs. 0.14 +/- 0.12 m/s, p = 0.0001), as well as in both the group with (0.25 +/- 0.12 vs. 0.13 +/- 0.13 m/s, p = 0.001) and the group without (0.39 +/- 0.02 vs. 0.15 +/- 0.12 m/s, p = 0.01) spontaneous echo contrast. In addition, left atrial and atrial appendage spontaneous echo contrast developed in 4 of 20 patients and increased in intensity in 3 of 20 patients in the immediate postcardioversion period. CONCLUSIONS: Organized left atrial appendage function returns in most patients immediately after cardioversion of atrial fibrillation. However, its function is impaired compared with that before cardioversion. Furthermore, spontaneous echo contrast increased in 7 (35%) of 20 patients after cardioversion. These observations suggest that stunned left atrial appendage function after cardioversion may predispose the chamber to thrombus formation, which may play a role in the mechanism involved in the occurrence of embolization after cardioversion. PMID- 8227793 TI - Repeated use of ablation catheters: a prospective study. AB - OBJECTIVES: The objective of this study was to provide insight into the time course of electrical, physical and mechanical changes in ablation catheters after each use that may affect the safety and efficacy of the ablation procedure. BACKGROUND: An increasing number of institutions are reusing deflectable ablation catheters. At present, there are no data concerning the safety of reusing ablation catheters. METHODS: Over a period of 1 year, 69 Webster/Mansfield deflectable catheters used in 336 ablation procedures were prospectively studied. An additional 18 new catheters were tested after multiple sterilizations only. The catheters were evaluated for electrical and physical integrity and mechanical capabilities. These include deflection at room temperature and 37 degrees C, shaft compression and buckling during deflection, tip craters, torquing ability, glue separation and tip attachment using a stereoscope at x30 magnification and electrical resistance for each electrode. After each use, the catheters were gas sterilized with ethylene oxide. RESULTS: The most common reasons for catheter rejection were tip electrode glue separation after 4.3 +/- 4.3 uses and loss of deflection after 5.0 +/- 3.3 uses. Electrical discontinuity between the catheter handle and electrodes was observed after 10.0 +/- 3.7 uses. There was no significant decrease in catheter torquing ability with repeated use. In this study the total estimated savings was $128,133, which includes the cost of catheter reprocessing. The reuse of Webster/Mansfield ablation catheters has not resulted in any major catheter failure or any major adverse clinical complications. CONCLUSIONS: On the basis of these observations, we believe that the Webster/Mansfield catheter can be reused an average of five times. It is strongly recommended that after each use catheters be carefully examined under appropriate magnification (x30) and that special attention be given to the ablation tip electrode. The catheters should also be tested for deflection and electrical integrity. PMID- 8227794 TI - Hemodynamic effects of oral sotalol during both sinus rhythm and atrial fibrillation. AB - OBJECTIVES: This study investigated the hemodynamic effects of oral sotalol during both sinus rhythm and paroxysmal atrial fibrillation. BACKGROUND: The hemodynamic effects of most antiarrhythmic drugs have been characterized in subjects during sinus rhythm. However, there are no data concerning these effects on the paroxysmal tachyarrhythmias. METHODS: In 17 patients with paroxysmal atrial fibrillation and without heart failure (aged 62 +/- 11 years, ejection fraction 51 +/- 4%), an electrophysiologic-hemodynamic study was performed twice. In the first study, hemodynamic variables were evaluated both during sinus rhythm and after the induction of atrial fibrillation. Sotalol (160 or 240 mg/day) was administered for 6 to 7 days and the study was then repeated with the same methods. RESULTS: The drug significantly diminished heart rate during both sinus rhythm and atrial fibrillation. During sinus rhythm, sotalol did not change systemic pressures and significantly increased left and right ventricular end diastolic, left and right atrial and pulmonary pressures. Cardiac index decreased, whereas stroke volume was unchanged after the drug. Ejection fraction and left ventricular end-diastolic and end-systolic volumes evaluated by echocardiography were unchanged after sotalol. During atrial fibrillation, the drug had less evident effects on cardiac function. Left ventricular end diastolic, left atrial and pulmonary pressures did not increase significantly. CONCLUSIONS: The hemodynamic changes induced by oral sotalol appear to be mainly related to an involvement of ventricular distensibility; this effect is less evident during atrial fibrillation than during sinus rhythm. In patients with paroxysmal atrial fibrillation without heart failure treated with oral sotalol, a recurrence of the tachyarrhythmia is hemodynamically well tolerated. PMID- 8227795 TI - Concepts of autoimmunity applied to idiopathic dilated cardiomyopathy. PMID- 8227796 TI - Acute effects of dobutamine on myocardial oxygen consumption and cardiac efficiency measured using carbon-11 acetate kinetics in patients with dilated cardiomyopathy. AB - OBJECTIVES: The aim of this study was to use positron emission tomography (PET) derived carbon (C)-11 acetate kinetics to determine the effects of dobutamine on oxidative metabolism and its effects on myocardial efficiency in a group of patients with dilated cardiomyopathy. BACKGROUND: Dobutamine is known to improve myocardial function but may do so at the expense of myocardial oxygen consumption, which could be a potential deleterious effect. Carbon-11 acetate kinetics correlate with myocardial oxygen consumption as shown in animal models. Combining these scintigraphic measurements of oxygen consumption with estimates of cardiac work results in a work-metabolic index, which reflects cardiac efficiency. METHODS: Eight patients with nonischemic dilated cardiomyopathy underwent dynamic PET imaging, echocardiography and hemodynamic measurements. Seven of these patients were also studied while receiving dobutamine. Direct measurements of myocardial oxygen consumption using coronary sinus catheterization were obtained with eight of the PET studies to validate C-11 acetate in patients with cardiomyopathy. RESULTS: The mean (+/- SD) C-11 clearance rate significantly increased with dobutamine from 0.105 +/- 0.027 to 0.155 +/- 0.023 min-1 (p = 0.001). Directly measured myocardial oxygen consumption had a linear relation to the mean C-11 clearance rate (r = 0.8, p = 0.018). Dobutamine was noted to significantly reduce systemic vascular resistance as well as the severity of mitral regurgitation. The work-metabolic index determined using hemodynamic variables and PET data increased from 2 +/- 0.7 x 10(4) to 2.6 +/- 0.6 x 10(4) (p = 0.04). Efficiency, estimated by employing the oxygen consumption to k2 relation, also increased from 13 +/- 4.5% to 16.9 +/- 6.4% (p = 0.04). CONCLUSIONS: Despite an increase in myocardial oxygen consumption, dobutamine led to an increase in work-metabolic index in patients with dilated nonischemic cardiomyopathy. Dobutamine reduced systemic vascular resistance and mitral regurgitation, suggesting that in this group of patients, it had important vasodilatory action in addition to its inotropic effects. The use of the C-11 acetate PET for determining myocardial oxygen consumption and estimating efficiency could potentially complement existing clinical measures of ventricular performance and may allow improved and objective evaluation of therapy in patients with heart failure. PMID- 8227797 TI - Regional specificity of peak hyperemic response in patients with congestive heart failure: correlation with peak aerobic capacity. AB - OBJECTIVES: The aim of this study was to compare peak reactive hyperemic blood flows in the forearm and calf of patients with congestive heart failure and in age- and gender-matched normal subjects. In addition, we attempted to correlate peak oxygen consumption with forearm and calf peak reactive hyperemic flows in the patients with heart failure. BACKGROUND: Disparate results have been reported regarding forearm peak reactive hyperemia in patients with congestive heart failure. Because training significantly increases peak reactive hyperemic flow in normal subjects, we hypothesized that in patients with congestive heart failure who curtail walking because of exertional symptoms, calf peak reactive hyperemic flow would be preferentially attenuated and that impairment of calf vasculature may correlate with peak oxygen consumption. METHODS: Forearm and calf blood flows were measured by venous occlusive plethysmography at rest and after 5 min of arterial occlusion in 46 patients with congestive heart failure and 7 age- and gender-matched normal subjects. Peak oxygen consumption was measured during graded exercise on a bicycle ergometer. RESULTS: Calf peak reactive hyperemic flow was lower in patients with congestive heart failure than in normal subjects (22 +/- 1 vs. 32.5 +/- 3.5 ml/min per 100 ml, p < 0.001), whereas forearm peak reactive hyperemic flows were similar in the two groups. Calf peak reactive hyperemic flow was linearly related to peak oxygen consumption (r = 0.58, p < 0.0001), but forearm peak reactive hyperemic flow was not. Forearm and calf peak reactive hyperemic flows were not related at rest or after 5 min of arterial occlusion in the patients with heart failure. CONCLUSIONS: Calf peak reactive hyperemic flow is reduced in patients with congestive heart failure, whereas forearm peak reactive hyperemic flow is identical to that of age- and gender matched normal subjects. Calf peak reactive hyperemic flow is linearly related to peak oxygen consumption in patients with congestive heart failure, but forearm peak reactive hyperemic flow is not. PMID- 8227798 TI - Regional remodeling and nonuniform changes in diastolic function in patients with left ventricular dysfunction: modification by long-term enalapril treatment. The SOLVD Investigators. AB - OBJECTIVES: The purpose of the present study was to assess the process of late regional remodeling and the changes in regional diastolic function at the base and apex of the left ventricle in patients with chronic systolic dysfunction. BACKGROUND: Remodeling has been suggested to play an important role in the progression of left ventricular dysfunction and heart failure. However, the regional difference in the process of late remodeling and its relation to diastolic function remain unclear. METHODS: In 32 patients with previous myocardial infarction and left ventricular ejection fraction < or = 35%, left ventricular hemodynamic and angiographic data were studied before and 1 year after randomization to conventional therapy with placebo (n = 12) or enalapril, 10 mg twice daily (n = 20). Left ventricular regional wall dynamics were analyzed in the basal and apical regions by the area method. RESULTS: In the placebo group, left ventricular end-diastolic and end-systolic regional areas increased significantly over time at the base but were unchanged at the apex. At the base, the diastolic left ventricular pressure-regional area relation shifted rightward and the regional stiffness constant decreased (6.9 +/- 4.3 to 5.0 +/- 3.1 x 10( 3) mm-2, p < 0.05), indicating an increase in regional distensibility. At the apex, however, the diastolic pressure-regional area relation shifted upward slightly, and the regional stiffness constant increased from 11.5 +/- 4.4 to 14.4 +/- 5.6 x 10(-3) mm-2 (p = 0.08). The regional peak filling rate was maintained at the base but decreased at the apex (1,014 +/- 436 to 762 +/- 306 mm2/s, p < 0.05); further, the changes in regional peak filling rate during follow-up were inversely related to the changes in the regional stiffness constant (r = -0.78, p < 0.001) at the apex. In contrast, in the enalapril group, end-diastolic and end systolic regional areas significantly decreased over time both at the base and at the apex. Diastolic pressure-regional area relations shifted leftward, but the regional stiffness constant and regional peak filling rate did not change significantly either at the base or at the apex. CONCLUSIONS: These findings suggest that in patients with severe systolic left ventricular dysfunction, there was a regional difference in the process of late remodeling between the base and apex of the left ventricle, which was associated with nonuniform changes in regional diastolic function in the placebo group. The data also suggest that the nonuniform progression of regional remodeling and diastolic dysfunction was prevented by long-term enalapril treatment. PMID- 8227799 TI - Coronary artery bypass grafting in severe left ventricular dysfunction: excellent survival with improved ejection fraction and functional state. AB - OBJECTIVES: The present study evaluated our experience with coronary artery bypass grafting in patients with severe left ventricular dysfunction. BACKGROUND: Despite the ominous prognosis of advanced ischemic cardiomyopathy, coronary artery bypass grafting in this setting remains controversial because of concerns over operative risk and lack of functional or survival benefit. METHODS: We analyzed the data of 83 consecutive patients (69 men, 14 women, aged 42 to 83 years [mean 66.8]) with a left ventricular ejection fraction < or = 30% who underwent isolated coronary artery bypass grafting (without aneurysmectomy, valve replacement or other open heart procedures) performed by one surgeon during a 6 year period. The ejection fraction ranged from 10% to 30% (mean 24.6%). Preoperatively, 49% of patients had angina, 52% had congestive heart failure (17% with pulmonary edema) and 30% manifested significant ventricular arrhythmia. The mean number of grafts was 2.7/patient. The internal mammary artery was used in 82% of grafts to the left anterior descending coronary artery. The intraaortic balloon pump was required therapeutically (for angina or pump failure) in 19% of patients and was prophylactically placed preoperatively in another 43% of patients. RESULTS: The hospital mortality rate was 8.4% (7 of 83). The mortality rate was 3.3% (2 of 61) in those patients who did not require admission to an intensive care unit immediately before operation. Canadian Cardiovascular Society angina class improved postoperatively by 1.9 categories and New York Heart Association congestive heart failure class by 1 category. Left ventricular ejection fraction (assessed postoperatively in 68 of 76 hospital survivors) improved from 24.6% preoperatively to 33.2% postoperatively (36% increase) (p < 0.001). At 1 and 3 years, respectively, all-cause survival was 87% and 80% and freedom from cardiac death was 89.8% and 84.5%. CONCLUSIONS: In patients with coronary artery disease and advanced ventricular dysfunction: 1) coronary artery bypass grafting can be performed relatively safely, 2) good medium-term survival is attained, 3) improvement in left ventricular function can be documented objectively after bypass grafting, 4) quality of life is improved (as reflected by improvement in anginal and congestive heart failure status), and 5) the internal mammary artery can safely be used as a conduit. The use of coronary artery bypass grafting is encouraged for this group of patients and may provide a viable alternative to transplantation in selected patients. PMID- 8227800 TI - Endothelium-dependent pulmonary artery responses in chronic heart failure: influence of pulmonary hypertension. AB - OBJECTIVES: The purpose of this study was to determine whether pulmonary artery responses to acetylcholine are abnormal in patients with chronic heart failure. BACKGROUND: Defective pulmonary artery endothelium-dependent responses have been observed in chronic heart failure models in animals. However, pulmonary artery endothelial responses in humans with chronic heart failure are unknown. METHODS: Twenty-two patients with chronic treated heart failure (12 with secondary pulmonary hypertension, Group I; 10 with normal pulmonary artery pressure, Group II) and 8 control patients constituted the study groups. Intravascular ultrasound measurements of pulmonary artery area just beyond the tip of an 8F infusion sheath were obtained in response to acetylcholine (10(-6), 10(-5) and 10(-4) mol/liter). The 10(-6) mol/liter infusion was repeated after methylene blue infusion. Indomethacin (5 micrograms/ml) was sequentially added to this combination in 17 patients. RESULTS: There were no significant differences among the three groups in vascular area responses to the lowest concentration (10(-6) and 10(-5) mol/liter) of acetylcholine, but the 10(-4) mol/liter infusion resulted in significant constriction in Group II patients (p < 0.05, analysis of variance [ANOVA]). Pretreatment with methylene blue in Group II also resulted in significant pulmonary artery vasoconstriction to even the 10(-6) mol/liter acetylcholine infusion (10.4 +/- 7.8% in Group II vs. 1.7 +/- 3.9% in the control group and 0.1 +/- 4.3% in Group I, p < 0.05, ANOVA). The addition of indomethacin resulted in reversal of the constriction in Group II patients. CONCLUSIONS: These responses indicate that the pulmonary artery endothelium may play a significant role in inhibiting vasoconstriction in patients with chronic heart failure who maintain normal pulmonary artery pressure. PMID- 8227801 TI - Noninvasive assessment of cardiac diabetic neuropathy by carbon-11 hydroxyephedrine and positron emission tomography. AB - OBJECTIVES: The purpose of this investigation was to evaluate the sympathetic nervous system of the heart by positron emission tomographic (PET) imaging in patients with diabetes mellitus with and without diabetic autonomic neuropathy. BACKGROUND: The clinical assessment of cardiac involvement in diabetic autonomic neuropathy has been limited to cardiovascular reflex testing. With the recent introduction of radiolabeled catecholamines such as carbon (C)-11 hydroxyephedrine, the sympathetic innervation of the heart can be specifically visualized with PET imaging. METHODS: Positron emission tomographic imaging was performed with C-11 hydroxyephedrine and rest myocardial blood flow imaging with nitrogen-13 ammonia. Three patient groups were studied, including healthy volunteers as control subjects, diabetic patients with normal autonomic function testing and diabetic patients with varying severity of autonomic neuropathy. Homogeneity of cardiac tracer retention as well as absolute tracer retention was determined by relating myocardial tracer retention to an arterial C-11 activity input function. RESULTS: Abnormal regional C-11 hydroxyephedrine retention was seen in seven of eight patients with autonomic neuropathy. Relative tracer retention was significantly reduced in apical, inferior and lateral segments. The extent of the abnormality correlated with the severity of conventional markers of autonomic dysfunction. Absolute myocardial tracer retention index measurements showed a 45 +/- 21% decrease in distal compared with proximal myocardial segments in autonomic neuropathy (0.069 +/- 0.037 min-1 vs. 0.13 +/- 0.052 min-1, p = 0.02). CONCLUSIONS: This study demonstrates a heterogeneous pattern of neuronal abnormalities in patients with diabetic cardiac neuropathy. The extent of this abnormality correlated with the severity of neuropathy assessed by conventional tests. Future studies in larger groups of patients are required to define the relative sensitivity of this imaging approach in detecting cardiac neuropathy and to determine the clinical significance of these scintigraphic findings in comparison with conventional markers of autonomic innervation. PMID- 8227802 TI - Silent myocardial ischemia: role of subclinical neuropathy in patients with and without diabetes. AB - OBJECTIVES: Silent myocardial ischemia is common in patients with diabetes. This study was designed to assess the role of subclinical autonomic impairment in diabetic patients with silent ischemia. BACKGROUND: Studies have suggested that silent ischemia is more common in diabetic patients with microvascular complications, but this has not been a consistent finding. METHODS: Twenty-two diabetic and 30 nondiabetic patients with proved coronary artery disease and a history of angina and ischemia on treadmill stress testing underwent clinical tests of autonomic function and measurement of 24-h heart rate variability. Diabetic patients with a history of microvascular complications were excluded. RESULTS: Although all 52 patients manifested ischemia during treadmill testing, only 36 patients experienced angina (angina group), whereas 16 did not (silent ischemia group). Diabetic and nondiabetic patients were similar in age (59 +/- 1 vs. 61 +/- 2 years, p = 0.56) and extent of coronary artery disease. However, clinical tests showed reduced parasympathetic function in the diabetic patients (Valsalva ratio 1.38 +/- 0.07 vs. 1.60 +/- 0.06; p = 0.007). Patients in the silent ischemia group were more often diabetic (33% vs. 63%, p = 0.05) and had prolonged time to ischemia on treadmill testing (200 +/- 20 vs. 271 +/- 20 s, p = 0.03). In addition, autonomic function was impaired in the silent group (supine/standing heart rate ratio 1.15 +/- 0.02 vs. 1.05 +/- 0.02, p = 0.002). Subgroup analysis showed that abnormalities of autonomic function were confined to the diabetic patients in the silent group. CONCLUSIONS: Despite the absence of overt microvascular complications, diabetic patients with silent exertional ischemia have evidence of significant autonomic impairment compared with findings in symptomatic patients. This difference is not seen in nondiabetic patients and indicates that subclinical neuropathy is an important cause of silent ischemia in patients with diabetes. PMID- 8227803 TI - Increased blood pressure and neural tone in the silent ischemia of hypertension: disparate effects of immediate release nifedipine. AB - OBJECTIVES: The aims of this study were 1) to evaluate the role of blood pressure and associated neural tonicity in ambient ischemia of a group of hypertensive patients with stable angina, and 2) to determine the efficacy of immediate release nifedipine therapy in controlling the total ischemic burden in both office-measured and ambulatory blood pressure. BACKGROUND: Low heart rate ischemia, as detected by Holter ambulatory electrocardiographic monitoring, suggests that reduced coronary flow is the major factor leading to ischemia. We previously found that 91% of the ischemic episodes in our hypertensive patients with stable angina were silent. METHODS: We measured plasma norepinephrine content during ischemic events from blood obtained from automatic pump withdrawal with the assistance of a real-time ST segment depression monitor. We then related the norepinephrine content to ischemic episodes assessed by 48-h Holter recording, blood pressure reading by ambulatory blood pressure monitoring and patients' diaries. Measurements were taken during the placebo period and immediate-release nifedipine therapy in 30 hypertensive patients (20 with and 10 without stable angina). RESULTS: More than half of the patients had ischemic episodes; 95% of these were silent. Ischemic episodes peaked in the early morning, and 55% occurred during routine sedentary activities. There was a 10% to 15% increase in heart rate at the onset of ischemia associated with a 30% higher plasma norepinephrine level. Seventy-five percent of patients had increased norepinephrine after nifedipine therapy. Nifedipine therapy controlled measured blood pressure but not 24-h ambulatory blood pressure. Ischemic episodes were reduced only in patients whose ambulatory blood pressure was controlled. CONCLUSIONS: The results suggest that increased neural tone at the time of the ischemic event may play a role in reducing coronary perfusion leading to silent ischemia. Nifedipine therapy (immediate release) was effective in control of ischemia only when both ambulatory and office-measured blood pressure were controlled. PMID- 8227804 TI - Silent ischemia after percutaneous transluminal coronary angioplasty: incidence and prognostic significance. AB - OBJECTIVES: The objective of this observational study was to assess the incidence and prognostic significance of silent ischemia after percutaneous transluminal coronary angioplasty. BACKGROUND: Apart from coronary angioplasty, prognosis of patients with silent ischemia is similar to that of patients with angina pectoris. However, similar data concerning silent ischemia associated with restenosis after coronary angioplasty are missing. METHODS: A consecutive series of 490 patients was investigated for asymptomatic ischemia on thallium-201 scintigraphy 6 months after successful coronary angioplasty. Repeat angiography was performed in a subgroup of patients with ischemia and repeat angioplasty was performed when clinically indicated. Patients were followed up for 2.2 +/- 0.8 years for cardiac events. RESULTS: Six months after coronary angioplasty, ischemia was present in 112 (28%) of 405 patients, and 60% of these 112 were asymptomatic. Ischemia was associated with significant stenosis in 97%; in contrast, results of exercise electrocardiography were negative in 74% of patients with scintigraphic ischemia and angiographic restenosis. The degree of restenosis was similar in patients with symptomatic or silent ischemia (80 +/- 16% vs. 81 +/- 21%). The long-term prognosis of patients with silent ischemia was remarkably similar to that of symptomatic patients. A worse outcome of symptomatic patients was found only if repeat coronary angioplasty for restenosis was considered a separate event (p < 0.01). Silent and symptomatic ischemia predicted an increased risk for recurrent ischemic events but not for death. CONCLUSIONS: Thus, absence of symptoms and negative findings on an exercise electrocardiogram may not reflect a good angioplasty result. In addition, silent ischemia due to restenosis after coronary angioplasty has a significant prognostic importance for recurrent symptomatic ischemic events that may be reduced by repeat angioplasty. PMID- 8227805 TI - Separate acquisition rest thallium-201/stress technetium-99m sestamibi dual isotope myocardial perfusion single-photon emission computed tomography: a clinical validation study. AB - OBJECTIVES: This study assessed the validity of a novel approach to myocardial perfusion scintigraphy that provides the opportunity to avoid the drawbacks of standard same-day rest/stress technetium-99m sestamibi myocardial perfusion studies by using separate-acquisition dual-isotope rest thallium-201 and exercise technetium-99m sestamibi single-photon emission computed tomography (SPECT). BACKGROUND: Standard same-day rest/stress technetium-99m sestamibi myocardial perfusion studies are cumbersome, associated with a potential decrease in perceived stress defect severity compared with thallium-201 due to the presence of rest technetium-99m sestamibi and may be unable to differentiate hibernating from infarcted myocardium. METHODS: The dual-isotope procedure was performed in 63 patients without previous myocardial infarction undergoing coronary angiography to evaluate sensitivity and specificity for coronary artery disease and in 107 patients with a low (< 5%) likelihood of coronary artery disease to evaluate normalcy rate. To validate defect reversibility, the dual-isotope SPECT study was compared with stress/rest technetium-99m sestamibi SPECT studies in a separate group of 31 patients with previous documented myocardial infarction who underwent a rest technetium-99m sestamibi study in addition to the dual-isotope SPECT study. RESULTS: In angiographic correlations, dual-isotope SPECT demonstrated high sensitivity for detecting patients with > or = 50% stenosis (91%, 55 patients) and > or = 70% stenosis (96%, 52 patients). In a small group of patients, high specificity was also observed (75% for < 50% stenosis [8 patients] and 82% for < 70% stenosis [11 patients]). A very high normalcy rate of 95% was also found. In the patient group assessed for defect reversibility, in zones with no previous myocardial infarction, segmental agreement for defect type between rest thallium-201 and rest technetium-99m sestamibi studies was 97% (kappa = 0.79, p < 0.001). In myocardial infarct zones, segmental agreement for defect type was 98% (kappa = 0.93, p < 0.001). Image quality was generally good to excellent. CONCLUSIONS: Our findings demonstrate that separate-acquisition dual-isotope myocardial perfusion SPECT is accurate for coronary artery disease detection, correlates well with rest-stress sestamibi studies for assessment of defect reversibility and results in good to excellent image quality. This approach provides an excellent method for the combined assessment of stress myocardial perfusion and myocardial viability. PMID- 8227806 TI - Left ventricular functional reserve in patients with syndrome X: evaluation by continuous ventricular function monitoring. AB - OBJECTIVES: The objective of this study was to evaluate cardiac functional reserve in patients with syndrome X. BACKGROUND: Syndrome X is characterized by stress-induced anginal pain and ST segment depression, normal findings on coronary angiography and normal left ventricular function at rest. Reduced coronary vasodilative reserve and abnormal myocardial lactate metabolism have been described in such patients. METHODS: To assess left ventricular functional reserve in patients with syndrome X, continuous radionuclide monitoring of left ventricular end-diastolic volume, end-systolic volume and ejection fraction was performed in 12 patients and 13 normal control subjects during supine bicycle ergometer exercise. RESULTS: In control subjects, end-diastolic volume increased at peak exercise from 100% to 106.5% (p < 0.01), end-systolic volume decreased from 39.1% to 22.6% (p < 0.01) and ejection fraction increased from 60.9% to 78.6% (p < 0.01). In patients with syndrome X, end-diastolic volume increased at peak exercise from 100% to 106% (p < 0.01), and end-systolic volume decreased at ST segment depression < or = 0.5 mm (the ST point) from 37% to 28.8% (p < 0.01) but increased at peak exercise to 44.7% (p < 0.01 vs. the ST point). Thus, ejection fraction increased at the ST point from 63% to 72.7% (p < 0.01) but decreased at peak exercise to 57.7% (p < 0.01 vs. the ST point and control subjects) in proportion to the degree of ST segment depression. In nine patients (75%), ejection fraction at peak exercise was lower than baseline values. All patients and control subjects showed a rapid ejection fraction increase just after exercise during the recovery period. The degree of ejection fraction "overshoot" in patients was similar to that in control subjects, but the interval from the end of exercise to the overshoot in patients was significantly longer than that in control subjects (118 vs. 65 s, p < 0.01). CONCLUSIONS: In patients with syndrome X subjected to exercise stress, left ventricular function remained normal before the onset of ST segment depression. Once ST segment depression appeared, left ventricular function deteriorated in proportion to the degree of depression, and reduced left ventricular function persisted into the recovery period. Continuous ventricular function monitoring is thus a useful predictor of reduced left ventricular functional reserve in patients with syndrome X. PMID- 8227807 TI - Prognostic significance of the pattern of aortic root dilation in the Marfan syndrome. AB - OBJECTIVES: The present study examines the incidence of aortic complications (dissection, marked dilation requiring surgery or progressive moderate to severe aortic regurgitation) and their relation to clinical features and aortic root morphology in patients with the Marfan syndrome. BACKGROUND: Considerable phenotypic variability exists in the Marfan syndrome, and the prospective prediction of the risk for aortic complications in individual patients remains elusive. METHODS: One hundred thirteen patients with the Marfan syndrome underwent anthropometric and echocardiographic evaluation and were followed-up for 49 +/- 24 (mean +/- SD) months. Aortic root dilation was defined as localized when confined to the sinuses of Valsalva (based on two-dimensional echocardiographic confidence limits utilizing age and body size) and generalized if dilation additionally involved the supraaortic ridge and proximal ascending aorta. RESULTS: Aortic root dilation was present in 80% of patients and was localized in 28% and generalized in 51%. Aortic complications occurred during follow-up in none of 23 patients with normal initial aortic size, in 2 (6%) of 32 patients with initially localized dilation and in 19 (33%) of 58 patients with generalized dilation (p < 0.0005). Complications were associated with larger initial aortic size (p < 0.00005), higher systolic blood pressure (p < 0.005), height (p < 0.05), aortic growth rate (p < 0.05) and older age (p < 0.01). The only independent predictor of aortic complications was initial aortic root size (p < 0.005). However, when aortic size, one of the indications for surgical referral, was excluded from analyses, the only independent predictor of aortic complications was generalized aortic dilation (p < 0.005). CONCLUSIONS: The present study indicates that generalized aortic root dilation is a potent marker of an increased risk for subsequent aortic complications in Marfan syndrome. PMID- 8227808 TI - Influence of collagen network on left ventricular systolic and diastolic function in aortic valve disease. AB - OBJECTIVES: The purpose of this study was to evaluate left ventricular structure function interplay in aortic valve disease. BACKGROUND: An increase in myocardial fibrosis has been demonstrated in aortic valve disease, but changes in the collagen network and their effect on ventricular function have not been defined. METHODS: Left ventricular structure was assessed from left ventricular endomyocardial biopsy specimens obtained in 32 patients with aortic valve disease (aortic stenosis in 25, aortic regurgitation in 7). Total collagen volume fraction, orthogonal collagen fiber meshwork (cross-hatching), endocardial fibrosis, muscle fiber diameter and volume fraction of myofibrils were determined by morphologic-morphometric evaluation. Control biopsy data were obtained from six donor hearts before transplantation. Eleven other patients with normal left ventricular function served as hemodynamic status control subjects. Left ventricular biplane cineangiography and high fidelity pressure measurements were carried out in all patients. Systolic function was assessed from ejection fraction. Diastolic function was evaluated by the time constant of relaxation, early and late peak filling rates and the constant of passive myocardial stiffness. Patients were assigned to three groups according to increasing severity of nonmyocyte tissue alterations. Group 1 comprised 10 patients with elevated total collagen volume fraction. Group 2 comprised 6 patients with normal total collagen volume fraction and the presence of increased cross-hatching or endocardial fibrosis, or both. Group 3 comprised 16 patients with elevated total collagen volume fraction and the presence of cross-hatching or endocardial fibrosis, or both. RESULTS: Muscle fiber diameter was increased in the three groups with aortic valve disease, whereas the volume fraction of myofibrils was comparable in all four study groups. Ejection fraction was depressed in groups 2 and 3 compared with the control group. The time constant of relaxation was prolonged in the three groups with aortic valve disease. No differences in early and late peak filling rate were observed in the four study groups, but the constant of myocardial stiffness increased in groups 2 and 3. CONCLUSIONS: In aortic valve disease, changes in collagen architecture are associated with altered systolic function and passive diastolic properties. The sole increase in total collagen volume fraction without a change in architecture leaves systolic and passive diastolic function unaltered. PMID- 8227809 TI - Active compression-decompression resuscitation: analysis of transmitral flow and left ventricular volume by transesophageal echocardiography in humans. Cardiopulmonary Resuscitation Working Group. AB - OBJECTIVES: This study was designed to test the hypothesis that active compression-decompression cardiopulmonary resuscitation increases transmitral flow and end-decompression left ventricular volume over levels achieved with standard manual cardiopulmonary resuscitation. BACKGROUND: Recently, cardiopulmonary resuscitation incorporating active compression and decompression of the chest has been demonstrated to improve hemodynamic status in a canine model and in humans after cardiac arrest. METHODS: The active compression decompression device was applied midsternum in five consecutive patients and results compared sequentially (in random order) with those of standard manual cardiopulmonary resuscitation. Both techniques were performed at 80 compressions/min with a 1.5- to 2-in. (3.8 to 5.1 cm) compression depth and a 50% duty cycle. Transesophageal echocardiographic data obtained in each patient included the velocity-time integral of transmitral pulsed Doppler recordings and two-dimensional images of the left ventricle in the long-axis view. Planimetric volume measurements of the left ventricle were obtained at both end-compression and end-decompression. RESULTS: No difference was observed in end-compression volume between the two techniques (p = 0.81). Increased end-decompression volume (active compression-decompression technique 81.3 +/- 12.5 vs. standard technique 69.4 +/- 10.8, p < 0.05), stroke volume (active compression-decompression technique 32.6 +/- 6.8 vs. standard technique 17.6 +/- 5.2, p < 0.05) and velocity-time integral of transmitral flow (active compression-decompression technique 15.8 +/- 4.3 vs. standard technique 7.8 +/- 2.3, p < 0.05) were found in the active compression-decompression group. The transmitral velocity-time integral was highly correlated with left ventricular stroke volume (r = 0.90). CONCLUSIONS: Improved transmitral flow, end-decompression left ventricular volume and stroke volume are seen with active compression-decompression resuscitation, suggesting a biphasic cardiothoracic cycle of flow. Active decompression of the chest appears to be a beneficial adjunct to standard cardiopulmonary resuscitation. PMID- 8227810 TI - Opacification and border delineation improvement in patients with suboptimal endocardial border definition in routine echocardiography: results of the Phase III Albunex Multicenter Trial. AB - OBJECTIVES: This study was designed to assess the safety and efficacy of intravenously administered sonicated human serum albumin for enhancing echocardiographic delineation of the left ventricular endocardium and improving assessment of wall motion in patients with incomplete depiction of noncontrast echocardiography. BACKGROUND: Echocardiographic regional wall motion analysis is impaired by incomplete endocardial definition in as many as 10% of patients. Sonicated human serum albumin is a stable contrast material that, unlike other agents, opacifies the left ventricle when administered intravenously. METHODS: One hundred seventy-five patients were enrolled at eight centers on the basis of incomplete echocardiographic endocardial depiction. Sonicated 5% human serum albumin, a stable preparation of air-filled microspheres (size range 1 to 10 microns), was administered intravenously in divided doses: 0.08 ml/kg body weight in all patients, followed by 0.14 and 0.08 ml/kg or a single dose of 0.22 ml/kg, depending on the result of the initial dose. Investigators and independent reviewers blinded to the protocol scored the echocardiograms for degree of left ventricular opacification and improvement of endocardial border depiction. RESULTS: Overall, 81% of patients had at least moderate left ventricular chamber opacification with at least one contrast dose, and endocardial definition was improved in 83%. In the subgroup with inadequate left ventricular opacification from the initial dose, a second, larger dose (0.22 ml/kg) improved endocardial depiction in 64%. No significant side effects occurred. CONCLUSIONS: In patients with incomplete echocardiographic endocardial definition, sonicated human serum albumin is a safe, effective contrast agent that, when administered intravenously, produces left ventricular chamber opacification, improves endocardial depiction and enhances regional wall motion analysis. PMID- 8227811 TI - Echocardiographic morphometry and geometry of the left ventricular outflow tract in fixed subaortic stenosis. AB - OBJECTIVES: This study was designed to identify, by echocardiography, morphometric abnormalities of the left ventricular outflow tract in children with fixed subaortic stenosis and to determine whether these abnormalities precede the development of subaortic obstruction. BACKGROUND: Fixed subaortic stenosis typically develops and progresses after the 1st year of life and is therefore often regarded as an acquired lesion. Although it has been speculated that there may be an underlying anatomic substrate, there are no data to support this hypothesis. METHODS: The size of the aortic annulus, mitral-aortic valve separation, aorto-left ventricular septal angle and degree of aortic override were determined in two groups of children. Group 1 comprised 35 patients with isolated subaortic stenosis noted on initial echocardiogram who were compared with an age- and weight-matched normal control group (Group 1A). Group 2 comprised 23 patients with ventricular septal defect or coarctation of the aorta, or both, who had no subaortic stenosis on initial echocardiogram but who developed it subsequently. This group was compared with an age-, weight- and lesion-matched control group (Group 2A). RESULTS: Compared with control subjects, patients with isolated subaortic stenosis had a significantly wider mitral-aortic separation ([mean +/- SD] 5.1 +/- 1.3 vs. 3.4 +/- 0.9 mm, p < 0.001), a steeper aortoseptal angle (131 +/- 6 degrees vs. 144 +/- 5 degrees, p < 0.001) and an exaggerated aortic override (p < 0.05). Similar differences were found on initial echocardiogram in Group 2 patients before development of subaortic stenosis: wider mitral-aortic separation (4.2 +/- 1.2 vs. 2.5 +/- 0.7 mm, p < 0.001), a steeper aortoseptal angle (132 +/- 7 degrees vs. 145 +/- 7 degrees, p < 0.001) and an exaggerated aortic override (p < 0.05). CONCLUSIONS: A left ventricular outflow tract malformation characterized by a wider mitral-aortic separation, an exaggerated aortic override and a steeper aortoseptal angle are present in children with ventricular septal defect or coarctation of the aorta, or both, who subsequently develop subaortic stenosis. These morphometric features can be used to identify by echocardiography patients who are at risk for developing fixed subaortic stenosis. PMID- 8227812 TI - Results of transcatheter closure of the patent ductus arteriosus with the Botallooccluder. AB - OBJECTIVES: Evaluation of the immediate and long-term efficacy of a novel method of transvenous closure of the patent ductus arteriosus was the purpose of this study. BACKGROUND: In 1981 at the First Moscow Hospital, a new method of transvenous patent ductus arteriosus closure with the Botallooccluder was developed. METHODS: Transvenous closure of the patent ductus arteriosus with the Botallooccluder was carried out in 273 patients. All patients were regularly followed up as outpatients at intervals of 3, 6 and 12 months after intervention. A total of 171 patients were examined at a mean of 3.2 +/- 1.1 years (range 1 to 8) after the procedure. Late evaluation with cardiac catheterization was performed in 62 patients (36%); 50 (29%) of these 62 also underwent right heart catheterization. RESULTS: Successful implantation was achieved in 266 patients (97%). The late follow-up findings testify to the favorable clinical course, with disappearance of the continuous murmur and regression of the electrocardiographic signs of ventricular overloading in the vast majority of patients. In no case after complete patent ductus arteriosus closure was a late ductus recanalization observed. No patient had even a minor degree of postprocedural lower extremity venous damage. In 29 patients (24%), the femoral artery was imaged by ultrasonography at late follow-up, without evidence of any vascular abnormality. CONCLUSIONS: The clinical data suggest high long-term efficacy and reliability of this technique. Introducer sheaths (12F to 16F outer diameter) did not cause any long-term entry site sequelae. PMID- 8227813 TI - Magnetic resonance volume flow and jet velocity mapping in aortic coarctation. AB - OBJECTIVES: Nuclear magnetic resonance (MRI) velocity mapping was used to characterize flow waveforms and to measure volume flow in the ascending and descending thoracic aorta in patients with aortic coarctation and in healthy volunteers. We present the method and discuss the relation between these measurements and aortic narrowing assessed by MRI. Finally, we compare coarctation jet velocity measured by MRI velocity mapping with that obtained from continuous wave Doppler echocardiography. BACKGROUND: The development of a noninvasive imaging method for morphologic visualization of aortic coarctation and for measurement of its impact on blood flow is highly desirable in the preoperative and postoperative management of patients. METHODS: Magnetic resonance imaging phase-shift velocity mapping was used to measure ascending and descending aortic volume flow in 39 patients with aortic coarctation and in 12 healthy volunteers. Magnetic resonance imaging was also used for anatomic and peak jet velocity measurements. The latter were compared with those available from continuous wave Doppler study in 40% of the patients. RESULTS: Whereas ascending aortic volume flow measurement did not show significant differences between the patient and healthy control groups, volume flow curves in the descending aorta did show significant differences between the two groups. Peak volume flow (mean +/- SD) was 10.6 +/- 5.3 liters/min in patients and 19.6 +/- 4.7 liters/min in control subjects (p < 0.001). Time-averaged flow was 2.5 +/- 0.9 liters/min in patients and 3.9 +/- 1.1 liters/min in control subjects (p < 0.05). The descending/ascending aorta flow ratio was 0.47 +/- 0.19 in patients and 0.64 +/- 0.08 in control subjects (p < 0.05). These variables correlate well with the degree of aortic narrowing. Peak coarctation jet velocity measured by MRI velocity mapping is comparable to that obtained from continuous wave Doppler study (r = 0.95). CONCLUSIONS: We established normal ranges for volume flow in the descending aorta and demonstrated abnormalities in patients with aortic coarctation. These abnormalities are likely to be related to resistance to flow imposed by the coarctation and could represent an additional index for monitoring patients before and after intervention. PMID- 8227814 TI - Effects of adjacent surfaces of different shapes on regurgitant jet sizes: an in vitro study using color Doppler imaging and laser-illuminated dye visualization. AB - OBJECTIVES: The present study was designed to estimate the influence of different shaped adjacent surfaces on regurgitant jets as assessed by color Doppler imaging and laser-illuminated dye optical visualization. BACKGROUND: Because color Doppler techniques provide real-time two-dimensional imaging of flow, the evaluation of valvular regurgitation by analysis of variance-encoded regurgitant jets by this method has been widely used in clinical studies. However, recent studies have demonstrated that color Doppler jet sizes are affected not only by several hemodynamic factors and instrument settings but also by the interaction between jets and adjacent wall surfaces. In clinical conditions, jets may interact with adjacent walls of variable shapes that might have different effects on the jet size. METHODS: An in vitro model was constructed consisting of a rigid, optically clear receiving chamber that had no outlet resistance and had a pulsatile pump ejecting through 1.5, 2.3 and 3.1 mm2 inflow orifices into the chamber. The surfaces were flat or smoothly and equally curved, convex and concave aluminum positioned at 0, 2 and 4 mm from and to the side of the inflow orifices. The pump was run with stroke volumes from 0.5 to 3.0 ml and with a pulse frequency of 70 beats/min. The echocardiographic and laser beams were aimed at the inflow orifice imaging jets perpendicular to the surfaces (vertical view) through the central plane of the jet flows. Maximal jet areas were measured by both color Doppler techniques and laser-illuminated dye visualization. RESULTS: Color Doppler study showed fair correlation between the jet areas and the stroke volumes (r = 0.83 to 0.99), but the jet sizes under different surface conditions were variable. All the surface jet areas at a jet-surface distance of 0 and 2 mm were smaller than free jet areas at the same stroke volume for both flat and convex surfaces (p < 0.001). Flow constraint by the concave surface resulted in the smallest jet areas (p < 0.001). The color Doppler jet areas on the curved surfaces were significantly smaller than the laser-illuminated dye visualization jet areas (p < 0.01 to 0.0001). However, at intermediate jet-surface distances (4 mm and sometimes 2 mm with higher velocity flows), jet interaction with the flat and especially with the convex surface resulted in larger jets. This effect was most pronounced on dye fluorescence studies because flow around these jets consisted mainly of low velocity vortical events with only partial surface adherence and these low velocity swirling flows were not well imaged by color Doppler technique. CONCLUSIONS: Our study suggests that the different-shaped adjacent surfaces with different degrees of flow alterations resulted in variable decreases in jet size and that color Doppler imaging could not encode and image the angled and low velocity swirling events well when jets flowed along the curved surfaces. These effects need to be taken into account when interpreting color Doppler images. PMID- 8227815 TI - Three-dimensional echocardiographic measurement of left ventricular volume in vitro: comparison with two-dimensional echocardiography and cineventriculography. AB - OBJECTIVES: This study was designed to compare three-dimensional echocardiography, two-dimensional echocardiography and cineventriculography for the purpose of measuring left ventricular volume in vitro. BACKGROUND: Three dimensional echocardiographic systems have been shown to be highly accurate in measuring the volumes of balloon phantoms. However, three-dimensional techniques have not been compared with standard two-dimensional echocardiography in vitro or with cineventriculography, the clinical standard for left ventricular volume measurement. METHODS: Excised porcine hearts were prepared with an internal latex sheath that could be filled and maintained with a known ("true") volume of liquid. Each heart was then imaged by cineventriculography, standard two dimensional echocardiography and three-dimensional echocardiography. Left ventricular volumes were calculated from 15 hearts at 25 volumes ranging from 50 to 280 ml by the following methods: 1) biplane cineventriculography using the area-length method; 2) two-dimensional echocardiography by the apical biplane method using a summation of discs algorithm in 15 cases and the single-plane, four-chamber method using a summation of discs algorithm in 10 cases; and 3) three-dimensional echocardiography using a polyhedral surface reconstruction volume computation algorithm based on multiple nonparallel, nonevenly spaced short-axis cross sections. RESULTS: Results were compared with true volume, and a nonparametric analysis of variance was performed. Both measurement bias (systematic error) and imprecision (random error) were assessed. All methods tended to underestimate the true volume (two-dimensional echocardiography -6.1 +/ 17.6%, three-dimensional echocardiography -4.7 +/- 5.0% and biplane cineventriculography -3.9 +/- 8.2%), although differences were not significant. Although there was a significant correlation between the magnitude of measurement bias and the size of the volume being measured for two-dimensional echocardiography and cineventriculography, the bias of three-dimensional echocardiography was fairly constant over the range of volumes. When bias was accounted for, two-dimensional echocardiography was significantly less precise than cineventriculography and three-dimensional echocardiography in terms of percent error (15.3 +/- 11.9%, 5.6 +/- 5.7% and 3.9 +/- 3.4%, respectively). CONCLUSIONS: Three-dimensional echocardiography using a polyhedral surface reconstruction algorithm for volume computation provides accuracy comparable to that of biplane cineventriculography in this in vitro model. Standard two dimensional echocardiographic volume computation is significantly less accurate than the other two methods. PMID- 8227816 TI - Measurement of left ventricular volume by three-dimensional echocardiography- present promise and potential problems. PMID- 8227817 TI - Experimentally created atrioventricular node reentrant tachycardia in the dog: evidence of a brake system for nodal reentry in the anterior interatrial septum. AB - OBJECTIVES: The purpose of the study was to investigate the hypothesis that nodal approaches of both anterior and posterior atrial input sites of the atrioventricular (AV) node contribute to part of the circuit of AV node reentrant tachycardia. Thus, tachycardia might be elicited by a premature atrial impulse that arrived at the AV node through one input site while blocked at another. BACKGROUND: Atrioventricular node reentrant tachycardia is the most common supraventricular tachycardia in humans, yet the exact pathway of the reentrant circuit is unknown. METHODS: In eight dogs, an operation that blocked atrial impulses from the anterior input site to the AV node was performed through a right thoracotomy with the inflow occlusion method. The right atrial free wall and the anterior atrial septum between the sinoatrial node and the AV node were completely divided, whereas the tissues within the triangle of Koch remained intact. Thus, atrial impulses were blocked from the anterior input site in the right atrium and the atrial septum and were conducted only through the left atrial free wall to the posterior atrial septum into the AV node. RESULTS: In a baseline electrophysiologic study before operation, dual AV conduction pathways were demonstrated in seven of eight dogs, but none of the seven had inducible AV node reentrant tachycardia. A repeat study 1 week postoperatively revealed that 1) both PR and AH intervals were prolonged during sinus rhythm (p < 0.01); 2) anterograde and retrograde conduction of the AV node showed no significant changes; and 3) AV node reentrant tachycardia was induced in four dogs (50%), of which three had sustained tachycardia. CONCLUSIONS: These results are compatible with the hypothesis that both nodal approaches of atrial input sites of the AV node contribute to part of the circuit of AV node reentrant tachycardia. They also confirm Moe's hypothesis of the existence of a brake system that prevents sustained AV node reentry. Our data suggest that the brake system is located in the anterior atrial septum. PMID- 8227818 TI - Changing requirements for training in cardiovascular diseases. PMID- 8227819 TI - President's page: doctors, placebos and a "letter from the King". PMID- 8227820 TI - Cytokines and acute cardiac rejection--the unfulfilled promise? PMID- 8227821 TI - Pulmonary embolism after cardiac surgery. PMID- 8227822 TI - Clinical consequences of angiotensin-converting enzyme inhibitor withdrawal in chronic heart failure: a double-blind, placebo-controlled study of quinapril. The Quinapril Heart Failure Trial Investigators. AB - OBJECTIVES: This study was performed to assess the efficacy, safety and clinical consequences of abrupt cessation of quinapril therapy in a placebo-controlled, randomized, double-blind withdrawal trial. BACKGROUND: Angiotensin-converting enzyme inhibitor therapy has assumed a pivotal role in the treatment of chronic heart failure. Quinapril hydrochloride, a nonsulfydryl angiotensin-converting enzyme inhibitor, has shown beneficial clinical effects in previous studies. METHODS: After > or = 10 weeks of single-blind quinapril therapy, 224 patients with New York Heart Association class II or III heart failure were randomized in double-blind fashion to continue quinapril (n = 114) or to receive placebo (n = 110) for 16 weeks. Changes in treadmill exercise time, New York Heart Association functional class, quality of life and symptoms of heart failure were assessed. RESULTS: Patients withdrawn to placebo had a significant deterioration in exercise tolerance (median change -16 s with placebo vs. +3 s with quinapril, p = 0.015). New York Heart Association functional class (p = 0.004) and quality of life were improved and signs and symptoms of congestive heart failure were lessened in those remaining on quinapril therapy compared with those receiving placebo. During double-blind treatment, 18 patients were withdrawn from the placebo group because of worsening heart failure compared with 5 patients withdrawn from quinapril treatment (p < 0.001). Rather than a precipitous deterioration of clinical status or early incidence of adverse events, withdrawal from quinapril was associated with steady worsening of heart failure, beginning 4 to 6 weeks after randomization to placebo. CONCLUSIONS: Quinapril is effective and safe for maintaining clinical stability in patients with moderate congestive heart failure. Withdrawal of quinapril from patients with heart failure results in a slow progressive decline in clinical status. PMID- 8227823 TI - Exercise hemodynamics during long-term implantation of a left ventricular assist device in patients awaiting heart transplantation. AB - OBJECTIVES: The goal of this study was to assess patients with end-stage heart disease after implantation of a left ventricular assist device at rest and during exercise compatible with activities of daily life. BACKGROUND: Mechanical circulatory assistance with a left ventricular assist device is an accepted therapy for bridging patients with end-stage heart disease to heart transplantation and has been proposed for long-term implantation. METHODS: Three patients (aged 37, 42 and 57 years) with end-stage heart failure required implantation of a pneumatically driven, asynchronous Thermedics left ventricular assist device while awaiting heart transplantation. All were assessed 1 month later during graded supine bicycle exercise (maximal work load 100 to 150 W). Detailed central hemodynamics, including continuous pulmonary artery oxygen saturation and oxygen consumption measurements, were obtained. Two of the patients also underwent upright treadmill exercise with oxygen consumption measurements. RESULTS: During supine bicycle exercise, the heart rate increased from 93 +/- 37 beats/min (95% confidence interval: mean +/- t0.025 x SE) at rest to 119 +/- 54 beats/min and left ventricular assist device rate increased from 82 +/- 47 to 109 +/- 55 beats/min. Oxygen consumption increased from 3.0 +/- 0.9 to 8.7 +/- 2.9 ml oxygen/min per kg body weight. Cardiac output increased from 6.0 +/- 4.4 to 9.6 +/- 7.1 liters/min, yielding an average exercise factor of 8.5 +/- 7.7 and an exercise index of 0.83 +/- 0.61. The patients assessed during treadmill exercise achieved a maximal oxygen consumption of 14.3 and 16.7 ml of oxygen/min per kg. No thromboembolic or other complications attributable to left ventricular assist device implantation occurred during the duration of support. All patients survived orthotopic heart transplantation and are doing well. CONCLUSIONS: Significant work loads compatible with activities of daily life and adequate exercise hemodynamics were demonstrated by these patients while awaiting heart transplantation. Definitive conclusions regarding the use of this device must be viewed as preliminary because only three patients were involved in this study and the failure rate may be as high as 71% (95% confidence interval of left ventricular assist device success as a bridge to transplantation 29.3% to 100%). Final conclusions regarding the safety and efficacy of the left ventricular assist device as a possible long-term circulatory support device must await results of larger multicenter trials in progress. PMID- 8227824 TI - Left ventricular function immediately after intravenous cocaine: a quantitative two-dimensional echocardiographic study. AB - OBJECTIVES: We tested the hypothesis that intravenous cocaine, in doses commonly self-administered in nonmedical settings, causes acute myocardial ischemia and left ventricular dysfunction. BACKGROUND: Cocaine-induced cardiac complications are responsible for a growing number of deaths in young people, but the mechanism by which cocaine induces these complications is unclear. METHODS: We performed 12 lead electrocardiography and quantitative two-dimensional echocardiography in 20 subjects before and after single intravenous doses of high dose cocaine (1.2 mg/kg body weight), low dose cocaine (0.6 mg/kg) and placebo. RESULTS: At 2 to 7 min after cocaine administration, the rate-pressure product was increased significantly from baseline (high dose 73%, low dose 63%, placebo 8%, p < 0.001 for either dose vs. placebo). During this time, electrocardiography demonstrated dose-related nonspecific changes (high dose in 14 of 20 subjects, low dose in 9 of 20 subjects, placebo in 2 of 20 subjects, p < 0.002 for either dose vs. placebo). In contrast, echocardiography showed that the frequency of hyperdynamic left ventricular wall segments doubled after high dose cocaine compared with placebo (34% [108 of 318] vs. 16% [51 of 319], respectively, p = 0.0001) but that there was no change in either left ventricular ejection fraction (high dose 66 +/ 9%, placebo 67 +/- 6%, p = NS) or wall motion score index (high dose 0.67 +/- 0.44, placebo 0.85 +/- 0.30, p = NS). CONCLUSIONS: We conclude that intravenous cocaine, in doses commonly self-administered in nonmedical settings, does not cause acute myocardial ischemia or left ventricular dysfunction. We speculate that cocaine-induced cardiac complications are caused by idiosyncratic coronary artery vasospasm, by exceptionally high dosages or by cocaine-induced coronary artery thrombosis. PMID- 8227825 TI - Comparison of carbon-11-acetate with fluorine-18-fluorodeoxyglucose for delineating viable myocardium by positron emission tomography. AB - OBJECTIVES: This study was designed to determine in patients with advanced coronary disease whether prediction of recovery of mechanical function after coronary revascularization could be accomplished more effectively by positron emission tomography (PET) with carbon-11 (11C)-acetate than by PET with fluorine 18 (18F)-fluorodeoxyglucose. BACKGROUND: Results of previous studies have demonstrated that preservation of myocardial oxidative metabolism (measured by PET with 11C-acetate) is necessary for recovery of systolic function after coronary revascularization. METHODS: Myocardial oxidative metabolism was quantified before revascularization in 34 patients by the analysis of the rate of myocardial clearance of 11C-acetate. Metabolism of glucose was assessed by analysis of uptake of 18F-fluorodeoxyglucose. Receiver operating characteristic curves for predicting functional recovery were derived for the measurements of oxidative metabolism and glucose metabolism. In addition, criteria for prediction of recovery of function based on measurements of oxidative metabolism and glucose metabolism were developed and compared. RESULTS: Analysis of receiver operating characteristic curves indicated that estimates of oxidative metabolism were more robust in predicting functional recovery than were estimates of glucose metabolism (p < 0.02). Moreover, threshold criteria with 11C-acetate exhibited superior positive and negative predictive values (67% and 89%, respectively) than did the criteria with 18F-fluorodeoxyglucose (52% and 81%, respectively), p < 0.01. In segments with initially severe dysfunction, estimates of oxidative metabolism tended to be more robust than estimates of glucose metabolism in predicting functional recovery. Moreover, in such segments, the threshold criteria with 11C-acetate tended to exhibit superior positive and negative predictive values (85% and 87%, respectively) than did the criteria with 18F fluorodeoxyglucose (72% and 82%, respectively), although statistical significance was not achieved. CONCLUSIONS: In patients with advanced coronary artery disease, the extent to which functional recovery can be anticipated after coronary revascularization can be delineated accurately by quantification of regional oxidative metabolism by PET with 11C-acetate. PMID- 8227826 TI - Scintigraphic and electrocardiographic evidence of silent coronary artery disease in asymptomatic hypertension: a case-control study. AB - OBJECTIVES: This study was conducted to determine the incidence of physiologically significant coronary artery disease in a group of asymptomatic high risk men with essential hypertension and to assess the validity of noninvasive tests in a subset of these patients undergoing coronary arteriography. METHODS: Two hundred twenty-six asymptomatic men (mean age 61 +/- 8 years) with essential hypertension and no clinical evidence of coronary artery disease but with at least one additional coronary risk factor were studied prospectively. Fifty age- and risk factor-matched normotensive subjects were evaluated as a control group. After a minimum of 4 days without medication, subjects underwent stress thallium-201 scintigraphy, exercise and 48-h ambulatory electrocardiography, and echocardiography. Coronary angiography was performed in a subset of 34 (40%) of 84 patients with one or more positive test results. RESULTS: A positive thallium-201 scintigram (18% vs. 6%; odds ratio 3.4, confidence interval 0.95 to 10.8, p = 0.056), exercise electrocardiograms (ECGs) (37% vs. 13%; odds ratio 4.1, confidence interval 1.5 to 11.2, p < 0.003) and ambulatory ECG (15% vs. 0%, p < 0.05) were more common in the hypertensive group than in the control group. In the cohort undergoing coronary angiography, thallium-201 scintigraphy was both sensitive and specific for epicardial atherosclerotic coronary disease (90% and 79%, respectively), but positive exercise and ambulatory ECGs occurred frequently in the absence of significant coronary stenoses. In the 39% of hypertensive patients who had mild to moderate left ventricular hypertrophy, positive exercise and ambulatory ECGs occurred at a higher rate. CONCLUSIONS: These findings suggest that physiologically significant coronary artery disease occurs more frequently in asymptomatic hypertensive men than in comparable normotensive control subjects. In the subgroup undergoing coronary arteriography, reversible scintigraphic defects were both sensitive and specific for diagnosing epicardial coronary artery disease, but exercise and ambulatory ECGs appeared to yield frequent false positive results, especially when left ventricular hypertrophy was present. These results indicate that patients with "silent" coronary artery disease can be identified among high risk hypertensive patients, but the appropriate application of such screening in clinical practice remains to be determined. PMID- 8227827 TI - Left ventricular hypercontractility and ST segment depression in patients with syndrome X. AB - OBJECTIVES: This study was designed to assess the relation between rest left ventricular function and exercise capacity in patients with syndrome X. BACKGROUND: Clinical observation has suggested that some patients with syndrome X have a high rest left ventricular ejection fraction. In this study we determined the relation between left ventricular ejection fraction and exercise capacity and the electrocardiographic (ECG) changes that develop on exercise. METHODS: The pattern of left ventricular function, exercise capacity and 24-h ambulatory ECG monitoring were studied in 37 patients (9 men, 28 women; mean age 52 +/- 7 years) with syndrome X (angina with normal coronary arteries and a positive exercise test result). All patients had normal findings on echocardiogram and rest ECG. All treatment was discontinued for > or = 48 h. Left ventricular ejection fraction was determined by computerized analysis of the left ventricular angiogram. In patients with syndrome X, exercise duration and heart rate were measured at 1-mm ST segment depression and at peak exercise. RESULTS: Left ventricular hypercontractility (ejection fraction > or = 80%) was observed in 12 patients (32%) (group 1), whereas 25 patients (68%) had normal left ventricular contraction (group 2). The time to 1-mm ST depression on exercise testing was significantly earlier in group 1 than in group 2 (5.13 +/- 1.03 vs. 10.76 +/- 0.63 min, respectively, p < 0.001). The magnitude of the ST segment depression at peak exercise was significantly greater in group 1 than in group 2 (2.03 +/- 0.2 vs. 1.33 +/- 0.05 mm, respectively, p < 0.001). The mean time for ST segment depression to normalize was significantly greater in group 1 than in group 2 (4.76 +/- 0.78 vs. 3.16 +/- 0.39 min, respectively, p < 0.05). Linear regression analysis of all patients with syndrome X showed a significant correlation between exercise duration and ejection fraction (r = 0.55, p < 0.001). The mean circadian variation of heart rate and episodes of ST segment depression on 24-h ambulatory ECG monitoring were similar in the two groups of patients. CONCLUSIONS: These findings indicate that approximately one third of patients with chest pain, normal coronary angiograms and a positive exercise test have left ventricular hypercontractility, and this is associated with the development of ST segment depression at a lower heart rate and work load and a longer time to normalization of ST segment depression after exercise. PMID- 8227828 TI - Beta-endorphins during coronary angioplasty in patients with silent or symptomatic myocardial ischemia. AB - OBJECTIVES: The aims of this study were to correlate beta-endorphin plasma levels and anginal pain in patients with ischemia induced by percutaneous transluminal coronary angioplasty and to detect eventual endorphin variations during balloon occlusion. BACKGROUND: The opioid system appears involved in the absence of pain occurring in silent myocardial ischemia. METHODS: Beta-endorphin plasma levels were measured 24 h before, just before, during and after coronary angioplasty (performed on the left anterior descending artery) in 53 men with documented coronary artery disease and exercise-induced myocardial ischemia. RESULTS: Group 1 (33 patients) reported symptoms; group 2 (20 patients) was asymptomatic during angioplasty. In these patients, the prevalence of exercise-induced silent ischemia was 57%. The occurrence of angina during exercise or angioplasty was related to the frequency of angina during daily life when patients were subgrouped. The severity and distribution of coronary artery disease did not differ between the two groups. During angioplasty, the number of balloon inflations and the inflation time and pressure were similar in symptomatic and asymptomatic patients. In each group, no short-term variability of baseline beta endorphin plasma levels was observed during 2 consecutive days. Corresponding beta-endorphin plasma levels (at baseline and during and after angioplasty) were significantly higher in Group 2. During balloon occlusion, the levels decreased significantly in the symptomatic group at the onset of angina but remained stable in the asymptomatic group. CONCLUSIONS: Methodologic variables and the severity of coronary artery disease did not influence the presence of symptoms during angioplasty-induced ischemia. Beta-endorphin plasma levels were higher and more stable in patients with silent ischemia during angioplasty, suggesting that opiate levels and their variation during ischemia are associated with individual attitude toward anginal pain. PMID- 8227829 TI - Prognostic value of an increase in fluorine-18 deoxyglucose uptake in patients with myocardial infarction: comparison with stress thallium imaging. AB - OBJECTIVES: This study was undertaken to evaluate the prognostic value of an increase in fluorine (F)-18 deoxyglucose uptake compared with clinical, angiographic and stress thallium findings in patients with myocardial infarction. BACKGROUND: Positron emission tomography (PET) imaging using F-18 deoxyglucose has been applied to assess tissue viability in patients with coronary artery disease. We hypothesized that patients with a myocardial segment with augmented F 18 deoxyglucose uptake are at high risk for a future cardiac event. METHODS: One hundred fifty-eight consecutive patients with myocardial infarction referred for F-18 deoxyglucose PET and stress thallium scans were studied. Follow-up was obtained in 84 patients at a mean interval of 23 months to investigate prognostic implications of radionuclide studies. RESULTS: Seventeen patients had a cardiac event during the follow-up interval. Univariate analysis showed that an increase in F-18 deoxyglucose uptake was the best predictor of a future cardiac event (p = 0.0006), followed by the number of stenosed vessels (p = 0.008). In the multivariate analysis, when an increase in F-18 deoxyglucose uptake was entered into the model, only angiographic variables had an independent prognostic value, whereas no other radionuclide variables showed significant prognostic value. Among patients who did not show redistribution, a future cardiac event was observed more often in patients with than in those without an increase in F-18 deoxyglucose uptake (p < 0.05). CONCLUSIONS: Thus, an increase in F-18 deoxyglucose uptake seemed to be the best predictor of a future cardiac event among all clinical, angiographic and radionuclide variables in this study of stable patients with myocardial infarction. Even when a stress thallium-201 scan does not show redistribution, those patients who have an increase in F-18 deoxyglucose uptake in a PET study may be at risk for a future cardiac event, and these patients may need aggressive treatment to prevent a future cardiac event. PMID- 8227830 TI - Are residual stenoses after excimer laser angioplasty and coronary atherectomy due to inefficient or small devices? Comparison with balloon angioplasty. AB - OBJECTIVES: The purpose of this study was to determine whether residual stenoses after excimer laser angioplasty and atherectomy were due to inefficient tissue ablation/removal or to undersized devices. BACKGROUND: Significant residual stenoses are commonly observed after use of laser and atherectomy devices. It is not known whether these residual stenoses are due to inefficient or undersized devices. METHODS: To determine the relative contribution of these factors, the minimal lumen diameter, percent diameter stenosis and normal reference diameter were measured immediately before and after coronary interventions in 696 lesions, including transluminal extraction atherectomy, high speed mechanical rotational atherectomy, excimer laser angioplasty and conventional balloon angioplasty. The ratio of the diameter of the device to the normal reference diameter (D/A, a measure of device sizing) and the ratio of the residual lumen diameter after use of the device to the device diameter (RLD/D, a measure of the efficiency of lumen enlargement) were calculated. RESULTS: Baseline diameter stenoses were similar for all interventions. The percent diameter stenoses were greater immediately after extraction atherectomy (60 +/- 21%), rotational atherectomy (54 +/- 23%) and excimer laser angioplasty (61 +/- 18%) compared with balloon angioplasty (26 +/- 12%, p < 0.001). The D/A ratio was smaller after extraction atherectomy (0.63 +/- 0.14), rotational atherectomy (0.59 +/- 0.17) and excimer laser angioplasty (0.51 +/- 0.11) compared with balloon angioplasty (1.05 +/- 0.13, p < 0.001). The RLD/D ratio was similar after extraction atherectomy (0.73 +/- 0.24) and balloon angioplasty (0.71 +/- 0.11) but was greater after rotational atherectomy (0.92 +/ 0.16, p < 0.001) and excimer laser angioplasty (0.85 +/- 0.30, p < 0.01) compared with balloon angioplasty. CONCLUSIONS: Residual stenoses after extraction atherectomy, rotational atherectomy and excimer laser angioplasty were more severe than after balloon angioplasty but were due to undersized devices (low D/A ratio), not to inefficient devices (low RLD/D ratio). Rotational atherectomy and excimer laser angioplasty were more efficient (higher RLD/D) than balloon angioplasty, whereas extraction atherectomy and balloon angioplasty were similar. PMID- 8227831 TI - Coronary vasoconstriction after angioplasty of total occlusions: relation to change in coronary perfusion pressure. AB - OBJECTIVES: This study evaluated the coronary vasomotor response after successful angioplasty of chronic total coronary occlusions and tested the hypothesis that the degree of distal vasoconstriction is dependent on the change in perfusion pressure. BACKGROUND: Although distal coronary vasoconstriction has been observed to occur frequently after successful percutaneous transluminal coronary angioplasty, little is known about this vasomotor response after angioplasty of chronic total occlusions. METHODS: Thirty patients with successful angioplasty of chronic total occlusions of the left coronary artery were studied. Quantitative coronary measurements were made at 0, 15 and 30 min after angioplasty and again after intracoronary nitroglycerin administration in coronary artery segments distal to the dilated lesion and in a control vessel. The change in distal perfusion pressure was defined as mean systemic arterial pressure minus residual mean trans-stenotic pressure gradient minus mean coronary wedge pressure. RESULTS: The distal segments showed greater vasoconstriction (vs. nitroglycerin) at 15 and 30 min compared with that at 0 min after angioplasty (32.3 +/- 2.2% and 35.2 +/- 2.5% vs. 12.5 +/- 1.8%, respectively, p < 0.005) and compared with control segments (32.3 +/- 2.2% vs. 12.1 +/- 2.0%, p < 0.005, and 35.2 +/- 2.5% vs. 12.5 +/- 2.0%, p < 0.005, respectively). The degree of vasoconstriction at 30 min in the distal segments was found to correlate closely with the change in distal perfusion pressure (r = 0.73, p < 0.001). CONCLUSIONS: Coronary distal vasoconstriction occurs frequently after successful angioplasty of chronic total occlusions and correlates closely with the change in coronary perfusion pressure. These findings support the hypothesis of reset epicardial coronary autoregulation in chronic hypoperfusion such that restoration of normal perfusion pressure after successful angioplasty may provoke reflex vasoconstriction. PMID- 8227832 TI - Fate of lesion-related side branches after coronary artery stenting. AB - OBJECTIVES: The aim of this study was to assess the immediate and long-term patency of lesion-associated side branches after coronary artery stenting. BACKGROUND: The possible adverse effects related to implantation of coronary stents are not completely known. An important potential complication of stenting is side branch occlusion due to mechanical obstruction or thrombosis. METHODS: Serial coronary angiography was performed in 153 patients (167 lesions) at baseline, after conventional balloon angioplasty, immediately after Palmaz-Schatz stent placement and at 6 months. The patency of side branches, where present, was analyzed at each of these points. RESULTS: Of 167 lesions stented, 57 stent placements spanned 66 side branches with a diameter > or = 1 mm. Twenty-seven (41%) of these side branches had > or = 50% ostial stenosis before standard balloon angioplasty. Six side branches became occluded after standard balloon angioplasty and remained occluded after stenting. Of the 60 side branches patent after conventional angioplasty, 57 (95%) remained patent immediately after stenting. All three side branches that became occluded after stenting had > or = 50% ostial stenosis at baseline. All 60 side branches, including the 3 initially occluded after stenting, were patent at 6-month follow-up. CONCLUSIONS: These findings demonstrate that 1) acute side branch occlusion due to coronary stenting occurs infrequently; 2) when side branch occlusion occurs, it is associated with intrinsic ostial disease; and 3) the patency of side branch ostia is well maintained at long-term follow-up. PMID- 8227833 TI - Noninvasive assessment of left internal mammary artery graft patency using duplex Doppler echocardiography from supraclavicular fossa. AB - OBJECTIVES: The purpose of this study was to clarify the usefulness of duplex Doppler echocardiography from the supraclavicular fossa for assessment of left internal mammary artery graft patency. BACKGROUND: A noninvasive method to assess coronary artery bypass graft patency would be useful for clinical diagnosis and long-term follow-up of graft outcome. METHODS: Duplex Doppler echocardiography from the supraclavicular fossa was performed in 56 consecutive patients who underwent postoperative cardiac catheterization studies, including quantitative angiography. All patients underwent coronary artery bypass graft surgery using the left internal mammary artery graft to the left anterior descending coronary artery. RESULTS: The left internal mammary artery graft and its flow were detected in 55 (98%) of the 56 patients with duplex Doppler echocardiography from the supraclavicular fossa. According to the quantitative angiographic data, the patients were assigned to three groups: group A (36 patients) with a normal left internal mammary artery graft (< 50% diameter stenosis), group B (9 patients) with intermediate (50% to 75% diameter) graft stenosis and group C (10 patients) with severe (> 75% diameter) graft stenosis. The diastolic/systolic peak velocity ratio was smaller in group C than in groups A and B (p < 0.05), but there was no significant difference between groups A and B. A diastolic/systolic peak velocity ratio < 0.6 predicted severe left internal mammary artery graft stenosis (> 75% diameter stenosis) with a sensitivity and specificity of 100% and 80%, respectively. The diastolic fraction of time-velocity integral was smaller in group C than in groups A and B (p < 0.05), but there was no significant difference between groups A and B. A diastolic fraction < 0.5 predicted significant left internal mammary artery graft stenosis (> 75% diameter stenosis) with a sensitivity and specificity of 90% and 100%, respectively. CONCLUSIONS: Duplex Doppler echocardiography from the supraclavicular fossa is useful for noninvasive assessment of left internal mammary artery graft patency. PMID- 8227834 TI - Diffuse reduction of myocardial beta-adrenoceptors in hypertrophic cardiomyopathy: a study with positron emission tomography. AB - OBJECTIVES: This study was conducted to determine the myocardial beta adrenoceptor density as a marker of sympathetic function in patients with hypertrophic cardiomyopathy and normal control subjects. BACKGROUND: Although some cases of hypertrophic cardiomyopathy are familial with an autosomal dominant pattern of inheritance, there remains a substantial proportion of cases in which neither a family history nor genetic abnormalities can be demonstrated. Additional abnormalities, both genetic and acquired, may be important in the phenotypic expression of this condition. Clinical features of the disease and metabolic studies suggest an increased activity of the sympathetic nervous system. METHODS: Eleven patients with hypertrophic cardiomyopathy, none of whom had previously received beta-blocking drugs, and eight normal control subjects underwent positron emission tomography to evaluate regional left ventricular beta adrenoceptor density and myocardial blood flow using carbon-11-labeled CGP 12177 and oxygen-15-labeled water as tracers. Plasma catecholamines were also measured. RESULTS: Mean (+/- SD) myocardial beta-adrenoceptor density was significantly less in the hypertrophic cardiomyopathy group than in the control group (7.70 +/- 1.86 vs. 11.50 +/- 2.18 pmol/g tissue, p < 0.001). Myocardial blood flow was similar in both groups (0.91 +/- 0.22 vs. 0.91 +/- 0.21 ml/min per g, p = NS). The distribution of beta-adrenoceptor density was uniform throughout the left ventricle in both groups. In the hypertrophic cardiomyopathy group, there was no correlation between regional wall thickness and myocardial beta-adrenoceptor density. There were no significant differences in either plasma norepinephrine or epinephrine concentrations between the two groups. CONCLUSIONS: There is a diffuse reduction in myocardial beta-adrenoceptor density in patients with hypertrophic cardiomyopathy in the absence of significantly elevated circulating catecholamine concentrations. This most likely reflects downregulation of myocardial beta-adrenoceptors secondary to increased myocardial concentrations of norepinephrine and is consistent with the hypothesis that cardiac sympathetic drive is increased in this condition. PMID- 8227835 TI - Purulent pericarditis: review of a 20-year experience in a general hospital. AB - OBJECTIVES: The purpose of this study was to review the features of purulent pericarditis in patients from a general hospital during a recent 20-year period. BACKGROUND: Although studies published from 1974 to 1977 suggested a changing spectrum for purulent pericarditis, this view has not been proved. METHODS: We retrospectively evaluated the records of 33 patients from one general hospital who had a diagnosis of purulent pericarditis during the period 1972 to 1991. All autopsy protocols from the same period were also reviewed. In 19 patients (group I), the condition was diagnosed during life; in 14 (group II), it was identified at autopsy. RESULTS: In group I, the possible sources of pericardial infection were identified in 17 patients; pneumonia (6 patients) was the most common source. Empyema was present in 10 patients; 15 had cardiac tamponade. The most common microorganisms were streptococci, pneumococci and staphylococci. Six patients developed constrictive pericarditis and required pericardiectomy. Three patients died, 1 patient was lost to follow up and 15 patients had a favorable outcome at a mean follow-up interval of 35 months. In group II, the clinical diagnoses included pneumonia (five patients) among other infections, with empyema in six patients. Purulent pericarditis was probably the direct cause of death in two patients. CONCLUSIONS: In our experience, the spectrum of purulent pericarditis has not changed in recent years. Many patients do not have the classical findings of pericarditis, and diagnosis is made only at autopsy or after tamponade has developed. Empyema remains a common predisposing condition. Purulent pericarditis is still a severe disease, but its prognosis is excellent in patients who can be discharged from the hospital. PMID- 8227836 TI - Changes in left and right atrial size after cardioversion of atrial fibrillation: role of mitral valve disease. AB - OBJECTIVES: The aim of this study was to examine the effect of cardioversion on left and right atrial volume in patients with chronic atrial fibrillation and to determine the influence of mitral valve disease on atrial size. BACKGROUND: Atrial enlargement is a common finding in atrial fibrillation and has been associated with an increased risk for embolic stroke. In addition, atrial enlargement may hamper long-term maintenance of sinus rhythm after cardioversion. METHODS: Forty-one patients with chronic atrial fibrillation (mean duration +/- SD, 45 +/- 62 months) underwent two-dimensional echocardiography before and 6 months after cardioversion to determine left and right atrial dimensions. Underlying heart disease was present in 26 patients: mitral valve disease in 12 (stenosis in 5, regurgitation in 5 and a combination in 2 patients) and other heart diseases in 14. Fifteen patients had lone atrial fibrillation. Patients with sustained sinus rhythm were compared with those who had a relapse of the arrhythmia 6 months after cardioversion. RESULTS: Six months after cardioversion, 28 patients still had sinus rhythm, whereas 13 patients had a relapse of the arrhythmia. In the 28 patients who had sinus rhythm after 6 months, left and right atrial volume decreased from a mean (+/- SD) 72.6 +/- 15.1 to 58.5 +/- 13.8 cm3 (-20%, p < 0.05) and from 68.7 +/- 14.6 to 58.6 +/- 11.6 cm3 (-14%, p < 0.05), respectively. Atrial dimensions also decreased significantly in the subgroup of patients with mitral valve disease. In contrast, no change in atrial size occurred in the 13 patients who had a relapse of atrial fibrillation. Left ventricular function did not change between the two echocardiographic studies, although New York Heart Association class improved in patients who had sinus rhythm after 6 months. CONCLUSIONS: Restoration of sinus rhythm reverts the process of left and right atrial enlargement in patients with chronic atrial fibrillation and mitral valve disease. Therefore, cardioversion may reduce the incidence of thromboembolic complications and prevent the arrhythmia from becoming refractory to medical therapy. PMID- 8227837 TI - Three-dimensional ultrasound imaging of the atrial septum: normal and pathologic anatomy. AB - OBJECTIVES: This study investigated the feasibility of producing three dimensional gray scale ultrasound images of the atrial septum to demonstrate normal and pathologic anatomy. BACKGROUND: Two-dimensional echocardiography is the principal technique used for imaging the atrial septum. Although the diagnostic accuracy of two-dimensional echocardiography is high, its capability for displaying complex three-dimensional relations is limited. METHODS: Three dimensional ultrasound images were reconstructed from tomographic images obtained during routine transesophageal echocardiographic examinations. Custom-made semi automatic algorithms for image enhancement, interpolation and segmentation were used to produce volumetric gray scale images. Volume-rendered displays of the atrial septum were generated for analysis. Sequential three-dimensional images were generated through the cardiac cycle and displayed cinematographically to permit assessment of motion. RESULTS: The three-dimensional images obtained from six patients clearly demonstrated normal and pathologic anatomy of the atrial septum, including atrial septal defects, atrial septal aneurysm and aortic valve ring abscess. The images could be manipulated electronically to demonstrate spatial relations and internal structural details. CONCLUSIONS: Three-dimensional gray scale reconstruction of ultrasound images obtained by transesophageal echocardiography is feasible. These images clearly demonstrate anatomic details and spatial relations. The gray scale images may be interactively manipulated to optimize the clinician's visualization of the atrial septum and its associated pathologic conditions. PMID- 8227838 TI - Left ventricular hypertrophy and mortality after aortic valve replacement for aortic stenosis. A high risk subgroup identified by preoperative relative wall thickness. AB - OBJECTIVES: We investigated the relation between the extent and pattern of left ventricular hypertrophy and surgical outcome in 54 patients undergoing aortic valve replacement for severe aortic stenosis. BACKGROUND: Previous work from our laboratory has demonstrated that a subgroup of patients, mostly elderly women with Doppler evidence of abnormal intracavitary flow acceleration, had an unexpectedly high in-hospital mortality rate after aortic valve replacement for aortic stenosis. We hypothesized that marked concentric hypertrophy, rather than the Doppler signal itself, was related to the poor outcome. METHODS: A retrospective analysis of the clinical, hemodynamic and echocardiographic data in patients who survived aortic valve replacement versus those who died in the hospital was performed. RESULTS: There were no differences between the 42 survivors and 12 nonsurvivors with regard to the clinical or hemodynamic variables. Of the echocardiographic variables analyzed, diastolic relative wall thickness was found to be significantly different between the two groups. Patients who died had significantly greater relative wall thickness (mean +/- SD) than those who survived (0.72 +/- 0.38 vs. 0.56 +/- 0.15, p = 0.04). Analysis by gender demonstrated that the relation between ventricular geometry and mortality held true only for women. CONCLUSIONS: We conclude that excessive ventricular hypertrophy, manifested as a markedly increased relative wall thickness, is associated with a significantly increased risk of postoperative mortality after aortic valve replacement for aortic stenosis. PMID- 8227839 TI - Amplitude-weighted mean velocity: clinical utilization for quantitation of mitral regurgitation. AB - OBJECTIVES: The purpose of this study was to determine the clinical usefulness of the amplitude-weighted mean velocity method for quantitation of mitral regurgitation. BACKGROUND: Amplitude-weighted mean velocity is a nonvolumetric method for calculating the mitral regurgitant fraction. Its previous validation at one center mandated an independent assessment of its usefulness and limitations. METHODS: In 56 patients with and 16 patients without mitral regurgitation, the regurgitant fraction was measured simultaneously by amplitude weighted mean velocity, quantitative Doppler study and quantitative two dimensional echocardiography. In 16 patients, multiple gain settings were used to determine the influence of this variable on amplitude-weighted mean velocity. RESULTS: In patients without regurgitation, amplitude-weighted mean velocity showed more scattering of regurgitant fraction (-18% to 23%) than Doppler (p = 0.016) or two-dimensional echocardiography (p = 0.022). The absolute value of regurgitant fraction was (mean +/- SD) 8 +/- 6%, 4 +/- 2% and 4 +/- 3%, respectively (p = NS). With increasing gain, the amplitude-weighted mean velocity mitral and aortic integrals increased, but the calculated regurgitant fraction remained unchanged. In patients with mitral regurgitation, significant correlation was found between amplitude-weighted mean velocity and Doppler study (r = 0.79, p = 0.0001) and between amplitude-weighted mean velocity and two dimensional echocardiography (r = 0.76, p = 0.0001) for calculated regurgitant fraction, but the standard error of the estimate (12%) was large. CONCLUSIONS: The amplitude-weighted mean velocity-calculated regurgitant fraction is gain independent, whereas the aortic and mitral integrals are gain dependent. Compared with Doppler and two-dimensional echocardiography, it shows more scattering of values in patients without regurgitation, but the methods correlate significantly in patients with mitral regurgitation. Amplitude-weighted mean velocity can be used as a simple adjunctive tool for comprehensive, noninvasive quantitation of mitral regurgitation. PMID- 8227840 TI - Contribution of mitral valve reserve capacity to sustained symptomatic improvement after balloon valvulotomy in mitral stenosis: implications for restenosis. AB - OBJECTIVES: To explain the discrepancy between the symptomatic status of patients and the hemodynamically calculated mitral valve area during long-term follow-up after mitral balloon valvulotomy, mitral valve orifice variability after dobutamine infusion was investigated in two groups of patients. BACKGROUND: A significant increase in aortic valve area with increased aortic transvalvular flow has been reported in patients with calcific aortic stenosis after aortic balloon valvulotomy. A similar phenomenon with regard to the mitral valve has not been studied in detail. METHODS: Group 1 comprised 10 patients (mean age 33 +/- 9 years) with untreated mitral stenosis. Group 2 comprised 29 consecutive patients (mean age 32 +/- 7 years) who underwent successful percutaneous mitral balloon valvulotomy 13 +/- 2 months before the study. RESULTS: After dobutamine infusion, heart rate and cardiac index increased significantly in both groups. The mean pulmonary artery pressure, mitral valve gradient and pulmonary capillary pressure remained unchanged in Group 2 but increased significantly in Group 1. The mean mitral valve area was significantly larger in Group 2 after dobutamine infusion than at baseline (1.9 +/- 0.5 vs. 2.4 +/- 0.6 cm2, p < 0.0001) but was unchanged in Group 1 (1.2 +/- 0.2 vs. 1.3 +/- 0.3 cm2, p = NS). The mean mitral valve area in seven patients in Group 2 (24%) was < or = 1.5 cm2 before dobutamine infusion (1.3 +/- 0.4 cm2), which was defined as restenosis. In five of these seven patients who had minimal or no symptoms, the mitral valve area increased significantly after dobutamine infusion (1.3 +/- 0.1 vs. 1.9 +/- 0.1 cm2). In the other two patients who were symptomatic, the mitral valve area did not change after dobutamine infusion. These two patients were identified as having "true" restenosis, and redilation of the mitral valve was performed in both. CONCLUSIONS: In patients who underwent mitral balloon valvulotomy, increased mitral valve reserve capacity contributed to symptomatic improvement on long-term follow-up. Dobutamine infusion may be helpful in detecting clinically significant restenosis. PMID- 8227841 TI - Cost-effectiveness of a smoking cessation program after myocardial infarction. AB - OBJECTIVES: The purpose of this study was to evaluate the cost-effectiveness of a smoking cessation program initiated after acute myocardial infarction. BACKGROUND: The value of allocating health care resources to smoking cessation programs after myocardial infarction has not been compared with the value of other currently accepted interventions. METHODS: A model was developed to examine the cost-effectiveness of a recently reported smoking cessation program after an acute myocardial infarction. The cost was estimated by considering the resources necessary to implement the program, and the effectiveness was expressed as discounted years of life saved. Years of life saved were estimated by modeling life expectancy using a single declining exponential approximation of life expectancy based on data from published reports. RESULTS: The cost-effectiveness of the nurse-managed smoking cessation program was estimated to be $220/year of life saved. In a one-way sensitivity analysis, the cost-effectiveness of the program remained < $20,000/year of life saved if the program decreased the smoking rate by only 3/1,000 smokers (baseline assumption 26/100 smokers), or if the program cost as much as $8,840/smoker (baseline assumption $100). In a two way sensitivity analysis, even if the cost of the program were as high as $2,000/participant, the cost-effectiveness of the program would be < $10,000/year of life saved so long as the program helped an additional 12 smokers quit for every 100 enrolled. CONCLUSIONS: Over a wide range of estimates of costs and effectiveness, a nurse-managed smoking cessation program after acute myocardial infarction is an extremely cost-effective intervention. This program is more cost effective than beta-adrenergic antagonist therapy after myocardial infarction. PMID- 8227842 TI - Routine hospital-based quit-smoking treatment for the postmyocardial infarction patient: an idea whose time has come. PMID- 8227843 TI - Junctional tachycardia: a useful marker during radiofrequency ablation for atrioventricular node reentrant tachycardia. AB - OBJECTIVES: The aim of this study was to evaluate junctional tachycardia as a useful marker during radiofrequency ablation for atrioventricular (AV) node reentrant tachycardia. BACKGROUND: Junctional tachycardia appears to be a response of the atrioventricular node to injury and is seen during both radiofrequency AV node ablation and slow and fast pathway ablation for AV node reentrant tachycardia. We hypothesized that junctional tachycardia heralding AV node block and that associated with slow or fast pathway ablation may have different characteristics that could be useful in preventing inadvertent AV block. METHODS: Characteristics of junctional tachycardia were examined after 59 radiofrequency ablation sessions in 53 consecutive patients with a mean age (+/- SD) of 41.6 +/- 16.5 years. Type 1 junctional tachycardia was followed by transient second- or third-degree AV block (n = 5) or permanent third-degree AV block (n = 1). Type 2 junctional tachycardia was followed by normal AV conduction (n = 53). RESULTS: Fifty-one patients had typical AV node reentrant tachycardia, and two patients had atypical tachycardia. Fast pathway ablation was attempted during 6 sessions and slow pathway ablation during 53 sessions. Patients underwent 15.3 +/- 10 radiofrequency applications, with a mean duration of 24 +/- 9.7 s. Junctional tachycardia was observed an average of 2.8 +/- 1.8 times per ablation session. Type 1 junctional tachycardia had a significantly faster rate than that of type 2 (cycle length 363 +/- 44 vs. 558 +/- 116, p < 0.001). In addition, type 1 junctional tachycardia was associated with predominantly ventriculoatrial block whereas type 2 was associated with predominantly 1:1 ventriculoatrial conduction (2 of 6 vs. 47 of 53 episodes, p < 0.05). CONCLUSIONS: We conclude that junctional tachycardia leading to AV block can be recognized by a faster junctional rate and ventriculoatrial block. This is a useful marker of impending AV block during slow and fast pathway ablation. PMID- 8227844 TI - Comparison of number of extrastimuli versus change in basic cycle length for induction of ventricular tachycardia by programmed ventricular stimulation. AB - OBJECTIVES: The purpose of this study was to examine the effects of varying basic cycle lengths in a programmed stimulation protocol if up to seven extrastimuli were available at each basic cycle length. BACKGROUND: There is no uniformly accepted protocol for induction of ventricular tachycardia. Most protocols limit the number of extrastimuli to two or three but use several basic cycle lengths. METHODS: Twenty-eight patients with coronary artery disease and documented spontaneous sustained ventricular tachycardia or ventricular fibrillation were studied. In the absence of antiarrhythmic drugs, each patient underwent three inductions of ventricular tachycardia/ventricular fibrillation using sinus rhythm or right ventricular pacing at 600 or 400 ms as the basic cycle length. Up to seven extrastimuli were allowed at each basic cycle length. RESULTS: The maximal yield of clinical tachycardia (96%) was identical for each basic cycle length and was achieved using a maximum of seven, five and four extrastimuli for sinus rhythm and 600 and 400 ms, respectively. A basic cycle length of 400 ms required fewer extrastimuli (2.4 +/- 0.7) to induce ventricular tachycardia/ventricular fibrillation than did 600 ms (2.7 +/- 1.1, p = 0.014) or sinus rhythm (3.4 +/- 1.2, p < 0.001). There was no significant difference in the cycle lengths of the induced ventricular tachycardia, incidence of induced ventricular fibrillation or requirement for direct current countershock. CONCLUSIONS: The use of an adequate number of extrastimuli obviates the need for multiple basic cycle lengths for induction of ventricular tachycardia and does not increase induction of unwanted ventricular fibrillation. If only one basic cycle length is used, the ease of inducibility can be quantified in terms of the number of extrastimuli required. Fewer extrastimuli were required for induction of ventricular tachycardia if a basic cycle length of 400 ms was used. These data favor the use of ventricular pacing at a basic cycle length of 400 ms with up to at least four extrastimuli as the standard stimulation protocol for induction of ventricular tachycardia. PMID- 8227845 TI - Bundle branch reentry: a mechanism of ventricular tachycardia in the absence of myocardial or valvular dysfunction. AB - OBJECTIVES: The aim of this study was to present bundle branch reentry as the mechanism of sustained ventricular tachycardia in the absence of myocardial or valvular dysfunction. BACKGROUND: Previous reports have documented the relation between structural heart disease and bundle branch reentrant ventricular tachycardia. Myocardial or valvular dysfunction has thus far been recognized as the only anatomic substrate for the development of this tachycardia. METHODS: Three patients with a wide QRS complex tachycardia underwent noninvasive and invasive cardiac evaluation and electrophysiologic studies to identify the substrate and mechanism of tachycardia. Catheter ablation of the right bundle branch using radiofrequency current was performed in each patient. RESULTS: The patients were all men (aged 54, 34 and 72 years) who presented with presyncope, palpitation and cardiac arrest, respectively. Electrocardiography during sinus rhythm revealed nonspecific intraventricular conduction delay in all three patients. Cardiac evaluation revealed no evidence of myocardial or valvular dysfunction in any patient. The baseline HV interval was prolonged in each patient (90, 100 and 75 ms, respectively). Programmed right ventricular stimulation initiated bundle branch reentrant tachycardia with typical left (three patients) and right (one patient) bundle branch block pattern. Catheter ablation of the right bundle branch using radiofrequency current abolished bundle branch reentry in all three patients. After 26-, 13- and 8-month follow-up periods, complete right bundle branch block persisted, and all three patients remained asymptomatic without antiarrhythmic drugs. CONCLUSIONS: Sustained bundle branch reentry can be a clinical arrhythmia in patients with no identifiable myocardial or valvular dysfunction except for isolated conduction abnormalities in the His-Purkinje system. This mechanism of tachycardia should be recognized during electrophysiologic evaluation, given the seriousness of this arrhythmia and the availability of the effective treatment. PMID- 8227846 TI - Prospective, randomized comparison of conventional and high dose loading regimens of amiodarone in the treatment of ventricular tachycardia. AB - OBJECTIVES: The purpose of this prospective randomized study was to compare the electrophysiologic effects of conventional and high dose loading regimens of amiodarone in patients with sustained ventricular tachycardia. BACKGROUND: Uncontrolled studies in which patients have been treated with an oral loading dose of 2 to 4 g/day of amiodarone have suggested that, compared with a conventional loading dose, this dosing regimen results in more rapid control of spontaneous ventricular tachycardia and ventricular tachycardia induced by programmed stimulation. METHODS: Patients in whom sustained monomorphic ventricular tachycardia was inducible by programmed stimulation and who were refractory to class I antiarrhythmic medications were randomly assigned to receive either a conventional (n = 15) or a high (n = 17) loading dose of amiodarone. The conventional dose consisted of 600 mg twice a day for 10 days. The high dose regimen consisted of 50 mg/kg body weight per day on days 1 to 3, 30 mg/kg per day on days 4 and 5 and 600 mg twice a day on days 6 to 10. An electrophysiologic test was performed in the baseline state and after 3 and 10 days of therapy. An adequate response to amiodarone was defined as the inability to induce ventricular tachycardia or the ability to induce only relatively slow (cycle length > or = 350 ms) hemodynamically stable ventricular tachycardia. RESULTS: After 3 days of therapy, 2 of 14 patients who received the conventional loading dose and 6 of 15 patients who received the high dose loading regimen had an adequate response to amiodarone (p = 0.08). After 10 days of therapy, four patients in each group had an adequate response to amiodarone (p = NS). Three patients who received the high dose and one patient who received the conventional dose of amiodarone had an adequate response after 3 days of therapy but not after 10 days of therapy. There were significant increases in the sinus cycle length, atrioventricular block cycle length, ventricular effective refractory period and ventricular tachycardia cycle length after 3 and 10 days of therapy compared with baseline values regardless of the dosing regimen. The extent of the effects of amiodarone on these variables after 3 and 10 days of therapy was similar with both dosing regimens. CONCLUSIONS: The therapeutic and electrophysiologic effects of conventional and high dose loading regimens of amiodarone do not differ significantly after 3 or 10 days of therapy. High oral loading doses of amiodarone do not offer any significant clinical advantage over a conventional loading dose of amiodarone for controlling ventricular tachycardia induced by programmed stimulation. PMID- 8227847 TI - Cardiac motion can alter proximal isovelocity surface area calculations of regurgitant flow. AB - OBJECTIVES: This study addressed the hypothesis that motion of the surface containing a regurgitant orifice relative to the Doppler ultrasound transducer can cause differences between actual flow rate and calculations based on the proximal flow convergence technique. BACKGROUND: In vitro studies quantitating regurgitant flow rate by proximal flow convergence have been limited to stationary orifices. Clinically, however, valve leaflets generally move relative to the ultrasound transducer during the cardiac cycle and can move at velocities important relative to the measured color aliasing velocities. The transducer therefore senses the vector sum of actual flow velocity toward the orifice and orifice velocity relative to the transducer. This can cause potential overestimation or underestimation of true flow rate, depending on the direction of surface motion. METHODS: The hypothesis was explored computationally and tested by pumping fluid at a constant flow rate through an orifice in a plate moving at 0 to 8 cm/s (velocities comparable to those described clinically for mitral and tricuspid annulus motion toward an apical transducer). RESULTS: Surface motion in the same direction as flow caused overestimation of the aliasing radius and calculated flow rate. Surface motion opposite to the direction of flow (typical for mitral and tricuspid regurgitation viewed from the apex or esophagus) caused underestimation of actual flow rate. The underestimation was greater for lower aliasing velocities (36 +/- 11% for 10 cm/s vs. 23 +/- 6% for 20 cm/s). Correcting for surface motion provided excellent agreement with actual values (y = 0.97x + 0.10, r = 0.99, SEE = 0.17 liters/min). CONCLUSIONS: Physiologic motion of the surface containing a regurgitant orifice can cause substantial differences between actual flow rate and that calculated by the proximal flow convergence technique. Los aliasing velocities used to optimize that technique can magnify this effect. Such errors can be minimized by using higher aliasing velocities (compatible with the need to measure the aliasing radius) or eliminated by correcting for surface velocity determined by an M-mode ultrasound scan. PMID- 8227848 TI - Limitation of myocardial infarct size in pigs with a dual lipoxygenase cyclooxygenase blocking agent by inhibition of neutrophil activity without reduction of neutrophil migration. AB - OBJECTIVES: The purpose of this study was to assess the effect of the dual cyclooxygenase-lipoxygenase blocking agent BW755C on the extent of myocardial infarction in the pig and to identify the mechanisms of any cardioprotective action of this drug. BACKGROUND: Activated neutrophils contribute to reperfusion injury after myocardial infarction and inhibition of neutrophil function can limit infarct size. METHODS: In 9 control and 10 study pigs pretreated with intravenous BW755C (10 mg/kg body weight) 30 min before coronary occlusion, ischemia was induced by a 50-min occlusion of the mid-left anterior descending coronary artery, followed by 3 h of reperfusion. Heart rate, arterial pressure, left ventricular end-diastolic pressure, the first derivative of left ventricular pressure (dP/dt) and regional myocardial blood flow were measured during control, occlusion and reperfusion periods. Infarct size was determined by histochemical staining; and myeloperoxidase activity, a marker for tissue neutrophil content, was assessed in normal and infarcted myocardium. The effect of BW755C on the function of isolated neutrophils stimulated with zymosan-activated serum was evaluated by measuring neutrophil degranulation, leukotriene B4 production, superoxide generation and chemotaxis. RESULTS: Hemodynamic function and regional myocardial blood flow were similar in control and BW755C-treated animals. BW755C significantly reduced myocardial infarct size compared with that in control animals, as measured by infarct/risk areas by histochemical staining (39 +/- 5% vs. 63 +/- 7%, p < 0.05). Myocardial myeloperoxidase activity was similar in normal, salvaged and infarcted areas in the control and treated groups, indicating that neutrophil accumulation in injured myocardium was unaltered by BW755C. However, this agent attenuated function of isolated, stimulated (zymosan activated serum) neutrophils. At a concentration of 0.03 mg/ml, BW755C inhibited degranulation (-46%), leukotriene B4 production (-48%) and superoxide generation (-74%), but there was minimal inhibition of chemotaxis in vitro. CONCLUSIONS: These findings demonstrate that myocardial infarct size can be reduced by selective inhibition of neutrophil cytotoxic activity without affecting neutrophil migration into injured myocardium. PMID- 8227849 TI - President's page: warning flags in the winds of health care change. PMID- 8227850 TI - Transesophageal echocardiography in right-sided endocarditis. PMID- 8227851 TI - Increased experimental atherosclerosis in cholesterol-fed rabbits exposed to passive smoke: taking issue with study design and methods of analysis. PMID- 8227852 TI - Is rubber elongation factor the major allergen of latex? PMID- 8227853 TI - Allergen skin testing. Board of Directors. American Academy of Allergy and Immunology. PMID- 8227854 TI - Characterization of a monoclonal antibody against latex protein associated with latex allergy. AB - Several hybridomas were produced against antigens extracted from the latex sap of Hevea brasiliensis. One of the monoclonal antibodies (LA-E3) reacted with antigens demonstrating binding to patient sera on crossed enzyme immunoelectrophoresis. This monoclonal antibody reacted with 2 of 10 glove extracts studied and with both Malaysian and Indian latex plant extracts. The antigens purified with monoclonal antibody affinity chromatography demonstrated specific IgE in the sera of patients with latex allergy as determined by ELISA. This monoclonal antibody can thus be utilized to obtain reliable antigens useful in the diagnosis of latex sensitivity, although additional antigens will likely be necessary to enhance sensitivity and specificity. PMID- 8227855 TI - Selection of Hymenoptera venoms for immunotherapy on the basis of patient's IgE antibody cross-reactivity. AB - BACKGROUND: Positive skin test results to multiple venoms in patients with Hymenoptera venom allergy may result from IgE antibody cross-reactivity among venom proteins. To avoid treatment with unnecessary and costly venoms, we have developed a venom RAST inhibition test that identifies individuals in whom a positive venom IgE RAST result is due to cross-reactive venom-specific IgE antibody. METHODS: Serum samples (n = 412) were collected over 5 years from patients with clinically characterized Hymenoptera venom allergy who had positive skin test results to more than one venom. Venom allergosorbent was added to serum containing IgE antivenin and buffer or 100 micrograms of homologous or heterologous venom. Bound IgE was detected with radiolabeled anti-human IgE. Intraassay variation less than 10% coefficient of variation and homologous venom inhibition greater than 80% were required for acceptance of data. A "cross reactivity index" (CRI) was computed as a ratio of percent inhibition produced by heterologous versus homologous venom. RESULTS: Of the 412 sera-venom combinations analyzed, 41 (10%) were excluded because of incomplete homologous venom inhibition. Of the 371 remaining sera, 82% (n = 305) were studied for IgE anti Polistes wasp venom (PWV) cross-reactivity with yellow jacket venom (YJV) and the other 66 for other venom specificity cross-inhibitions. Of the serum samples tested, 36.4% (111 of 305) contained IgE anti-PWV venom of which the binding to solid-phase PWV was inhibited with soluble YJV to a level that produced CRIs greater than 95%. We believe that this constitutes complete inhibition and demonstrates exclusively YJV cross-reactive antibody in these samples. The remaining 63.6% had CRIs from 0% to 95%, indicating IgE specific for a spectrum of unique and cross-reactive PWV allergens. Only 4.3% (13 of 305) had CRIs less than 5%, which is consistent with IgE restricted to PWV unique allergens. The degree of the IgE anti-PWV inhibition to solid-phase PWV by YJV was independent of the IgE anti-PWV level. CONCLUSIONS: This study shows that one third of patients with Hymenoptera venom allergy evaluated with positive YJV- and PWV reactive IgE in the skin and/or serum were identified as candidates for exclusion of PWV from their immunotherapy regimen because their IgE anti-PWV was more than 95% cross-inhibitable with YJV. Cost analysis of the venom RAST inhibition test and a conventional 5-year Hymenoptera venom immunotherapy program indicates that this serologic evaluation is cost-effective. PMID- 8227856 TI - Analysis of cross-reactivity between sunflower pollen and other pollens of the Compositae family. AB - The sera of 20 patients with Compositae pollen allergy were investigated for the presence of IgE antibodies reacting against sunflower pollen by means of RAST and immunoblotting studies. Thirteen IgE-binding bands were detected with molecular weights ranging from 14.4 to 94 kd. Two of these bands, with molecular weights of 24 and 25 kd, contained major allergens that reacted strongly with 100% (24 kd) and 95% (25 kd) of the sera, respectively. Cross-reactivity between sunflower and other Compositae pollens (mugwort, marguerite, dandelion, golden rod, and short ragweed) was revealed by RAST and immunoblotting inhibition experiments. Mugwort pollen exhibited the greatest degree of allergenic homology (cross-reactivity) with sunflower pollen, whereas at the other end of the spectrum, short ragweed showed less cross-reactive epitopes. PMID- 8227857 TI - Prospective study of risk factors in natural rubber latex hypersensitivity. AB - Five hundred sixty-nine subjects routinely underwent skin prick tests for latent sensitization to latex. The study of risk factors included skin tests to inhalant allergens, to diagnose atopy, and a questionnaire aimed at revealing frequent exposure to latex such as the wearing of gloves, multiple surgical procedures, or urinary catheterization. The subjects were categorized into five groups: group I, subjects with no risk factor (n = 272); group II, nonatopic subjects exposed to latex (n = 73); group III, atopic subjects not exposed (n = 180); group IV, exposed atopic subjects (n = 44); and group V, subjects with a history of intraoperative anaphylactic shock caused by latex (n = 13). Twenty-five subjects had spina bifida and were in either group II (14 subjects) or group IV (11 subjects). The questionnaire identified a probable allergy to latex in 18 subjects: 16 cases were confirmed by skin test, but responses were not informative in 23 patients who were sensitive to latex. Positive prick tests to latex were obtained in 0.37% of group I, 6.85% of group II, 9.44% of group III, and 36.36% of group IV. Of the children with spina bifida, 32% had positive skin test results. As risk factors, atopy and exposure were synergistic. We recommend predictive prick tests not only in children with spina bifida but also in any atopic subject or in any patient with a history of frequent exposure to latex. Latex could be considered a habitual allergen. The use of latex urinary catheters should be avoided in patients who are catheterized on a daily basis. PMID- 8227858 TI - Troleandomycin in the treatment of difficult asthma. AB - BACKGROUND: For difficult asthma, treatment is aimed at improving airway obstruction and minimizing adverse effects of systemic corticosteroids. The combination of troleandomycin (TAO) with methylprednisolone (MP) reportedly has a beneficial steroid-sparing effect on difficult asthma. METHODS: To test the steroid-sparing effect of TAO, 14 subjects with severe corticosteroid-dependent asthma were studied before and during treatment with MP and TAO. RESULTS: Treatment with MP and TAO resulted in a clear reduction in respiratory symptoms, asthmatic attacks, corticosteroid and hospitalization requirements, improvement in pulmonary function tests, and a remarkable decrease in peak expiratory flow rate circadian variability when compared with the period before TAO treatment (with corticosteroids). Treatment with MP and TAO was fairly well tolerated. Only a reduction (-5.2%, p < 0.01) in bone mineral content and an increase in plasma glucose levels (from 81.7 to 94.3 mg/dl, p < 0.05) were found, in comparison with pre-TAO values. One subject discontinued MP-TAO treatment because of a mild but persistent increase in serum alanine aminotransferase. In an evaluation of the clinical and pulmonary function test results, eight of the 14 subjects were responders and six were nonresponders. During follow-up three subjects discontinued TAO and consequently had an asthmatic attack after 4, 7, or 15 days, respectively. CONCLUSIONS: This study confirms that treatment with MP and TAO has a beneficial effect in a subgroup of severely steroid-dependent asthma patients. PMID- 8227859 TI - Skin responses to bradykinin, kallidin, and [desArg9]-bradykinin in nonatopic and atopic volunteers. AB - BACKGROUND: Kinins are potent vasoactive oligopeptides that may act as mediators in a variety of inflammatory diseases of the skin by interacting with specific receptors designated B1 and B2. In this study we have investigated the structure activity relationship of intradermally injected bradykinin, kallidin (lysine bradykinin), and [desArg9]-bradykinin in atopic (n = 8) and nonatopic (n = 8) subjects. METHODS: On two separate occasions, each separated by a week, subjects randomly underwent intradermal challenge with incremental doses (0.5, 5, and 50 nmol) of either the B1-agonist [desArg9]-bradykinin, the B2-agonists bradykinin or kallidin, or vehicle placebo. In a separate randomized double-blind study we have also examined the effect of an orally administered antihistamine, terfenadine, on kinin-induced wheal and flare responses and their repeatability in a group of nine volunteers. The skin responses were monitored objectively by measurement of wheal and flare areas. RESULTS: Both bradykinin and kallidin induced a dose-dependent increase in wheal and flare areas in all subjects studied. Although the effects of the two lowest doses (0.5 and 5 nmol) of [desArg9]-bradykinin on skin responses were indistinguishable from those of placebo, this kinin at the highest dose administered (50 nmol) caused a significant increase in wheal and flare areas in all subjects studied. No difference could be identified in the skin responses to kinins between atopic and nonatopic subjects. In addition kinin-induced cutaneous responses were not altered by pretreatment with terfenadine. CONCLUSIONS: These in vivo structure activity studies suggest that in human beings the skin responses to kinins may be compatible with the stimulation of B2 receptors, which is unrelated to histamine release from cutaneous mast cells. PMID- 8227860 TI - The rubber elongation factor of rubber trees (Hevea brasiliensis) is the major allergen in latex. AB - BACKGROUND: Allergy to latex-containing articles is becoming more and more important because it can result in unexpected life-threatening anaphylactic reactions in sensitized individuals. METHODS: A protein of 58 kd with an isoelectric point of 8.45 was purified from raw latex and from latex gloves and identified as the major allergen, completely blocking specific IgE antibodies in the serum of latex-sensitized subjects. The allergen is a noncovalent homotetramer molecule, in which the 14.6 kd monomer was identified, by amino acid composition and sequence homologies of tryptic peptides, to be the rubber elongation factor found in natural latex of the Malaysian rubber tree. RESULTS: Competitive immunoinhibition tests showed that the starch powder covering the finished gloves is the airborne carrier of the allergen, resulting in bronchial asthma on inhalation. The purified allergen can induce allergic reactions in the nanogram range. CONCLUSION: The identification of the allergen (Hev b I) may help to eliminate it during the production of latex-based articles in the future. PMID- 8227861 TI - Isolation and characterization of a major allergenic component (gp55) of Aspergillus fumigatus. AB - IgE class antibodies specific for antigens in a water-soluble extract of Aspergillus fumigatus (strain NHL-5759) were analyzed by immunoblotting with sera from patients with allergic bronchopulmonary aspergillosis. All the sera tested were reactive with a major 50 to 60 kd protein in the extract. This allergen, designated gp55, was purified by gel filtration and preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigen was found to be present in the water-soluble extract in the form of a complex composed of approximately eight molecules of gp55. The carbohydrate and phosphate content of the purified antigen were 23.1% and 0.46%, respectively. The molar ratio of mannose to galactose residues was 2.76:1, and the protein was glycosylated predominantly with N-linked oligosaccharides. The serologic activity of the gp55 antigen was abolished by treatment with nonspecific protease (Pronase) but not by treatment with sodium metaperiodate or endoglycosidases. Thus the major antigenic site of the glycoprotein is located within its peptide moiety. The antigen itself displayed no chymotryptic or tryptic activity. The amino acid sequence of the 20 N-terminal residues of the antigen (ATPHEPVFFSWDAGAVTSFP) is different from that of any other protein previously reported. PMID- 8227862 TI - Allergens in Hymenoptera venom. XXV: The amino acid sequences of antigen 5 molecules and the structural basis of antigenic cross-reactivity. AB - The complete amino acid sequences have been determined by solid-phase protein sequencing for eight different vespid venom antigen 5 molecules. These include five species of yellow jackets, Vespula squamosa, V. flavopilosa, V. germanica, V. pensylvanica and V. vidua, representing all three species groups; two variants from the European hornet, Vespa crabro; and a species of paper wasp, Polistes fuscatus, from a second subgenus. The new sequences were compared with the seven previously published sequences from yellow jackets, hornets, and wasps, and to that of Solenopsis invicta 3 allergen from imported fire ant venom. These comparisons provided structural evidence to support the observed high degree of cross-reactivity among the antigens of the common group of yellow jackets and among those of the two common North American subgenera of paper wasps studied. The antigen 5 of V. squamosa and of V. vidua were significantly different from those of the vulgaris group. Common features that could generate immunologic cross-reactivity were seen among the antigen 5 molecules of hornets of both genera and among those of yellow jackets, hornets, and paper wasps. The imported fire ant allergen has only minimal conserved areas in common with the vespid allergens, which explains the lack of observed IgE cross-reactivity. These results provide the structural basis for the cross-reactivity patterns observed in clinical practice and suggest that the commercial extracts of yellow jacket and paper wasp could be prepared with fewer carefully selected species. PMID- 8227863 TI - Intranasal steroids inhibit seasonal increases in ragweed-specific immunoglobulin E antibodies. AB - We performed two seasonal studies to evaluate the effect of continuous treatment with intranasal steroids, beginning approximately 1 week before the appearance of ragweed pollen, on the level of ragweed-specific IgE antibodies in serum. In both studies the control groups showed the anticipated rise in ragweed-specific IgE antibodies after the ragweed season. In the first study, employing aqueous beclomethasone dipropionate (168 micrograms twice daily), no rise occurred in serum ragweed IgE after seasonal exposure and the level actually decreased in eight of 12 treated subjects. In the second study, with triamcinolone acetonide (220 micrograms twice daily), the expected rise in ragweed IgE antibody was also reduced, although less dramatically, probably as a result of the lower potency of the dose delivered. Our studies not only support the benefits of intranasal steroids in the treatment of seasonal allergic rhinitis but also suggest that specific IgE production may be down-regulated by their continuous use, which may alter the subsequent clinical course of the disease. PMID- 8227864 TI - Analysis of nasal secretions during experimental rhinovirus upper respiratory infections. AB - BACKGROUND: To determine the underlying mechanisms for rhinovirus-induced nasal secretions, nasal lavage fluids were analyzed during experimental rhinovirus infections. METHODS: Twenty patients with allergic rhinitis and 18 nonallergic control subjects were inoculated with rhinovirus type 39. Nasal lavage was performed before and on days 2 through 7 after viral inoculation, and the lavage fluids were assayed for proteins and mast cell mediators. RESULTS: The secretion of total protein and both plasma proteins (albumin and IgG) and glandular proteins (lactoferrin, lysozyme, and secretory IgA) increased after rhinovirus inoculation. Analysis of the specific protein constituents revealed that nasal secretions during the initial response to the rhinovirus infection were predominantly due to increased vascular permeability. Allergic subjects tended to have fewer symptoms and more vascular permeability than control subjects, and increased histamine secretion after rhinovirus inoculation was more frequently seen in the allergy group. CONCLUSION: Nasal secretions found early in the course of a viral upper respiratory infection are due to increased vascular permeability, whereas glandular secretions predominate later in the infection. PMID- 8227865 TI - Effect of rhinovirus 39 infection on cellular immune parameters in allergic and nonallergic subjects. AB - Patients with allergic rhinitis (AR), compared with nonallergic persons, have been reported to respond differently to a variety of stimuli, some of which are immunologic in nature. This study compared the systemic cellular immune responses to experimental rhinovirus (RV) 39 challenge in RV-39-seronegative AR (n = 20) and nonallergic (n = 18) subjects. Peripheral blood was obtained before, 4 or 7 days after, and 23 days after RV-39 intranasal challenge and assayed for the number and function of various white blood cells. All subjects were infected, as manifested by viral shedding in nasal secretions or seroconversion. RV-39 induced marked changes from baseline values in both immune cell number and functions. Compared with nonallergic subjects, AR subjects manifested different responses for the following parameters: (1) numbers of total white blood cells and lymphocytes (smaller increases on day 4), (2) helper/suppressor T cell ratio (absence of an increase on day 7 and presence of an increase on day 23), (3) number of IL-2 receptor-positive suppressor T cells (presence of a decrease on day 7), (4) natural killer (NK) cell numbers (absence of an increase on day 4 and presence of increases on days 7 and 23), (5) NK/T cell ratio (absence of an increase on day 4 and a decrease on day 7), (6) NK cell activity (a blunted decrease on day 7 and absence of a decrease on day 23), and (7) RV-39-induced lymphocyte proliferation (exaggerated increase on day 4). The results show that intranasal challenge with RV-39 induced RV-39-specific and nonspecific systemic cellular immune responses and a unique immunologic response pattern in AR subjects. PMID- 8227866 TI - Antecedent features of children in whom asthma develops during the second decade of life. AB - In this report we compare 36 subjects in whom asthma was first diagnosed between the ages of 10 and 20 with 297 control subjects. All subjects were studied at age 5 to 9. Among the subjects who acquired a new diagnosis of asthma (NDA), the diagnosis was usually preceded by lower respiratory tract symptoms (31 of the 36 subjects had respiratory symptoms or a diagnosis of rhinitis or chronic bronchitis before asthma developed). Among those tested, more of those with NDA had positive allergy skin test results (56.5%) before diagnosis than control subjects (29.6%; p < 0.05), and the subjects with NDA had higher levels of serum IgE than control subjects (mean log serum IgE = 2.27 in subjects with NDA, 1.76 in control subjects; p < 0.05). Pulmonary function tests revealed no significant differences in the groups before diagnosis. Using logistic regression, we determined that wheezing, cough, a diagnosis of chronic bronchitis, and a positive allergy skin test result were independent risk factors for asthma. When combinations of variables were used, subjects with wheezing and a positive allergy skin test result, cough and a positive test result, and also those with a prior diagnosis of chronic bronchitis alone were at highest risk of a subsequent diagnosis of asthma. PMID- 8227867 TI - Comparative performance of five commercial prick skin test devices. AB - Five commercially available devices for performing prick skin testing were compared for reproducibility, patient acceptance, occurrence of false-negative skin test results, and size distribution of reactions at the negative control sites. Reproducibility of skin testing with 10 mg/ml histamine base, as expressed by coefficient of variation, was similar. However, a clear range of trauma to the skin was produced by the devices. This trauma was least with the Hollister-Stier and ALK Laboratories lancets, intermediate for the bifurcated needle by either prick or puncture, and greatest for the Multi-Test and DermaPIK devices. The more traumatic devices produced larger mean wheals and more frequent and large reactions at saline control sites, and were less acceptable to subjects. However, except for the Multi-Test device, they less often yielded false-negative responses. It is proposed that for each device a different size of wheal must be produced at the allergen site to have confidence that it exceeds the control site. The wheal size necessary for 99% specificity were as follows: Hollister Stier lancet, 2 mm; ALK lancet, 3.0 mm; bifurcated needle prick, 4.0 mm; bifurcated needle puncture, 4.5 mm; Multi-Test device, 5.0 mm; and DermaPIK device, 5.5 mm. An additional observation was the presence of a significant gradient of reaction size on the back to both histamine and allergen (p < 0.0001), with the smallest reactions in the upper third and the largest in the lower third of the back. PMID- 8227868 TI - Inhibition of neutrophil functions by human immunoglobulin E. AB - Incubation of human neutrophils with human immunoglobulin (Ig) E caused dose dependent inhibition of adhesion, phagocytosis, secretion of myeloperoxidase, and oxygen radical production. The concentrations of IgE that caused 50% inhibition of adhesion, phagocytosis, and secretion were 2 +/- 0.3, 2.16 +/- 0.21, and 1.95 +/- 0.28 ng/ml, respectively. Oxidase activation as measured by luminol-dependent chemiluminescence by the receptor-mediated N-formyl-methionyl-leucyl phenylalanine, phorbol 12-myristate 13-acetate, or the particulate stimulus Staphylococcus aureus was inhibited by IgE with concentrations causing 50% effect of 1.2 +/- 0.13, 1.09 +/- 0.16, and 0.6 +/- 0.09 ng/ml, respectively. IgE also inhibited oxygen consumption rate and cytochrome c reduction with similar K0.5 values. The effect of IgE was unlikely to be due to nonspecific cytotoxicity because trypan blue exclusion test and the cytoplasmic marker lactate dehydrogenase revealed that the cells retained their viability after IgE treatment. Similar or higher concentrations of IgG invoked either no inhibition or a slight enhancement of neutrophil functions. Pretreatment of neutrophils with IgG failed to affect the IgE-induced inhibition. Because the effect of IgE occurs at concentrations less than those reported in hyperimmunoglobulinemia E, we propose that direct inhibition of neutrophil functions may underlie the pathogenesis of recurrent infection associated with hyperimmunoglobulinemia E. PMID- 8227869 TI - The effectiveness of benzyl benzoate and some essential plant oils as laundry additives for killing house dust mites. PMID- 8227870 TI - Airway responsiveness to adenosine 5'-monophosphate in smokers and nonsmokers with atopic asthma. PMID- 8227871 TI - Can we change diet to improve health? PMID- 8227872 TI - Effective weight loss strategies for black women. PMID- 8227873 TI - Who's paying for nutrition services? Billing, coding, and reimbursement trends. PMID- 8227874 TI - Legislative initiatives for health care reform: an analysis of five proposals, including the Clinton plan. PMID- 8227875 TI - Protein intake and the calcium economy. PMID- 8227876 TI - Interdisciplinary perceptions of the dietitian's legal responsibility for lethal dietary prescription errors for patients with end-stage renal disease. AB - OBJECTIVE: To determine intergroup and intragroup consensus for interdisciplinary perceptions of dietitians' legal responsibility for lethal dietary prescription errors. DESIGN: Survey research was conducted by a mail questionnaire containing four hypothetical cases in which the dietitian provides a lethal amount of dietary potassium for an anephric patient. SETTING: Of the 90 New York State end stage renal disease facilities contacted, 64 returned one or more questionnaires. SUBJECTS/SAMPLES: Usable questionnaires were tabulated from 51 registered dietitians, 39 registered head nurses, and 43 physician-medical directors. MEASURES: Consensus was recognized when 60% or more of the responses occurred in one extreme collapsed and dichotomized category of "yes" or "no" regarding the dietitian's legal responsibility. STATISTICAL ANALYSES PERFORMED: Intergroup and intragroup consensus levels were compared by frequency, mean, standard deviation, and Scheffe test of differences. RESULTS: The triad groups met varying levels of consensus on all four scenarios, with dissenting opinions by nurses on two scenarios. The dietitians and the physician-medical directors had mirror-image perceptions that the dietitian was not legally accountable for two cases of following foreseeably lethal dietary prescriptions "as written." Eight dietitians viewed the dietitian as legally nonculpable for causing a patient's death because of the dietitian's sole incompetence or carelessness in making a serious calculation error. Nurses met the highest levels of agreement and perceived the dietitian to be legally accountable for all four cases. The nurses, in contrast to the dietitians and physician-medical directors, held opinions of dietitian legal responsibility as consistent with the standard of care expectations of prudent health care professionals defined by professional malpractice jurisprudence. APPLICATIONS/CONCLUSIONS: The data indicate that to protect both the patient and the professional, there is a notable need for greater understanding dietetics jurisprudence. PMID- 8227877 TI - Calcium intakes of Mexican Americans, Cubans, Puerto Ricans, non-Hispanic whites, and non-Hispanic blacks in the United States. AB - OBJECTIVE: To compare dietary calcium intakes from food in Mexican Americans, Cubans, Puerto Ricans, non-Hispanic whites, and non-Hispanic blacks aged 11 through 74 years. DESIGN: Population survey data from the Hispanic Health and Nutrition Examination Survey and the second National Health and Nutrition Examination Survey were used to calculate calcium intake from a single 24-hour recall. These data were compared by age and sex between the five population groups. Food sources of calcium in the three Hispanic groups were also examined using 24-hour recall data. SUBJECTS: The sample consisted of 11,773 non-Hispanic whites, 1,728 non-Hispanic blacks, 4,739 Mexican Americans, 1,076 Cubans, and 1,835 Puerto Ricans. MAIN OUTCOME MEASURES: Mean calcium intake, percentage intake of Recommended Dietary Allowance, and, for Hispanics, food sources of calcium. STATISTICAL ANALYSES: Means were compared within age and sex groups between the five population groups using a t test. RESULTS: Calcium intakes from food in the three Hispanic groups were similar to intakes of non-Hispanic whites and higher than intakes of non-Hispanic blacks. Although dairy foods were the main sources of calcium for Hispanics, corn tortillas were important calcium sources among Mexican Americans. Women consumed less calcium than the Recommended Dietary Allowance in all age and racial or ethnic groups. APPLICATIONS: When assessing calcium intakes of the three Hispanic groups, ethnic differences in food sources of calcium need to be considered. Efforts to increase calcium intake in Hispanics also need to account for ethnic differences. PMID- 8227878 TI - Food pattern, diet quality, and related characteristics of schoolchildren in New York State. AB - OBJECTIVES: To examine the food patterns and diet quality of elementary schoolchildren in New York State (outside of New York City) and to determine sociodemographic characteristics correlated with diet quality. DESIGN: A nonquantitative 24-hour recall administered to students and a brief questionnaire completed by parents. SAMPLE: 1,797 second and fifth graders (51% of those asked) in 51 randomly selected schools (46% of those asked) in New York State outside of New York City. STATISTICAL ANALYSES: Multiple regression analyses, chi 2, and t tests. RESULTS: On the day they were surveyed, 40% of students did not eat vegetables, except for potatoes or tomato sauce; 20% did not eat fruit; 36% ate at least four different types of snack foods, and 16% of fifth graders did not eat breakfast. Children who ate a school lunch ate significantly more dairy foods and fruits and vegetables, and fewer snack-foods than those who brought lunch from home. Fifth graders ate significantly more snack foods and were more likely to skip breakfast than second graders; boys had lower food-group pattern scores than girls; children of lower socioeconomic status had less diverse diets but ate less snack foods than children of higher socioeconomic status; children with single parents were more likely to skip breakfast and to eat fewer vegetables than those with two parents; and children with mothers employed outside the home had less diverse diets than those with mothers at home. APPLICATIONS/CONCLUSIONS: The findings indicate that nutrition interventions are clearly needed for this age group and that targeted messages should be based on sociodemographic characteristics. PMID- 8227879 TI - Vitamin A as a hormone: recent advances in understanding the actions of retinol, retinoic acid, and beta carotene. AB - Within the past few years, much has been learned about the metabolism and actions of vitamin A and the carotenoids. This article reviews the biochemical and cellular events in retinoid metabolism that lead to production of retinoic acid, an active metabolite of vitamin A. Retinoic acid functions in a hormone-like manner to regulate the expression of a number of genes. Beta carotene is now under study as an anticancer agent and for its possible beneficial effects in a number of chronic diseases. Current recommendations for carotene intake exceed the usual daily intake nearly fourfold. PMID- 8227880 TI - Integrating research and grant writing in an undergraduate dietetics program. AB - The Council on Research of The American Dietetic Association considers research by dietitians a necessity. A number of authors conclude that the link between research and practice needs to begin with the educational experience. We report on a model for integrating research and grant proposal writing in the undergraduate dietetics curriculum that has been used successfully for 6 years in a coordinated program. Students carry out research projects so that they learn teamwork, read research articles more critically, become more skilled with statistical analysis, and learn how to seek funding for their work. This model could easily be adopted by educational programs. Having such research and writing experience as students may stimulate continued research in the practice setting. PMID- 8227881 TI - Model for selecting quality standards for a salad bar through identifying elements of customer satisfaction. AB - Continuous quality improvement is the new requirement of the Joint Commission on Accreditation of Healthcare Organizations. This means that meeting quality standards will not be enough. Dietitians will need to improve those standards and the way they are selected. Because quality is defined in terms of the customers, all quality improvement projects must start by defining what customers want. Using a salad bar as an example, this article presents and illustrates a technique developed in Japan to identify which elements in a product or service will satisfy or dissatisfy consumers. Using a model and a questionnaire format developed by Kano and coworkers, 273 students were surveyed to classify six quality elements of a salad bar. Four elements showed a dominant "must-be" characteristic: food freshness, labeling of the dressings, no spills in the food, and no spills on the salad bar. The two other elements (food easy to reach and food variety) showed a dominant one-dimensional characteristic. By better understanding consumer perceptions of quality elements, foodservice managers can select quality standards that focus on what really matters to their consumers. PMID- 8227882 TI - Are we really fixing up the food supply? AB - Twenty years ago, in response to a general conviction that nutrition education had failed, many nutrition professionals supported fortification of the food supply with vitamins and minerals. Although the nutrition problems of most concern today are the result of macronutrient excesses, not micronutrient deficiencies, once again there is an effort to correct these problems by altering the food supply. This article reviews the arguments used to justify widespread micronutrient fortification in the years after World War II. Micronutrient fortification was undertaken without sufficient evidence of need; however, evidence is lacking that it was ineffective nutritionally. Assuming that the goal of the dietetics profession is to teach the public to select healthful diets, the energy-reduced fat substitutes, designer starches, nonnutritive sweets, and various fibers now being added to the food supply are unlikely to help nutritionists achieve that goal. PMID- 8227883 TI - Modification of dietary oxalate and calcium reduces urinary oxalate in hyperoxaluric patients with kidney stones. PMID- 8227884 TI - Use of the Delphi research technique to test a decision model in foodservice systems: a case study in food production. PMID- 8227885 TI - Influences of eating patterns on change to a low-fat diet. PMID- 8227886 TI - Attitudes toward infant feeding among adolescent mothers from a WIC population in northern Louisiana. PMID- 8227887 TI - Eating quality of muffins, cake, and cookies prepared with reduced fat and sugar. PMID- 8227888 TI - Position of the American Dietetic Association: vegetarian diets. PMID- 8227889 TI - Profile of depressive symptoms in younger and older medical inpatients with major depression. AB - OBJECTIVE: To examine and compare the profile of cognitive symptoms, somatic symptoms, and somatic complaints in younger and older medical inpatients diagnosed with major depressive disorder (MDD). DESIGN: Cross-sectional cohort. PATIENTS AND SETTING: Male patients aged 20-39 years (n = 116) and aged 70-102 years (n = 332), consecutively admitted to the medical and neurological services of a VA medical center, underwent psychiatric evaluations for depression. MEASUREMENTS: Depressive symptoms were measured using the observer-rated Hamilton Depression Scale and the self-rated Geriatric Depression Scale using an "inclusive" approach (symptoms rated regardless of etiology). Data on 12 common somatic complaints were also collected. Clinical diagnoses of MDD were made by a psychiatrist based on the Diagnostic Interview Schedule and other psychiatric and physical health data. RESULTS: After controlling for severity of medical illness, functional status, and alcohol use, symptoms that discriminated MDD best among younger men were loss of interest, feelings of guilt or of being a burden, suicidal thoughts, and depressed mood (primarily cognitive and affective symptoms). Among older men, loss of interest, insomnia, suicidal thoughts, and hypochondriasis most strongly differentiated depressed from non-depressed patients (mixture of cognitive and somatic symptoms); fatigue, weight loss, genital symptoms, and somatic anxiety were weakly related or unrelated to MDD. Number of somatic complaints (headache, dizziness, palpitations, etc.) helped differentiate patients with MDD; although equally common among depressed young and older patients, a distinct pattern of complaints emerged in each age group. CONCLUSIONS: Cognitive symptoms of depression can help to distinguish medical inpatients with and without MDD. In older adults, however, somatic symptoms cannot be ignored and may be more important for diagnosing MDD than previously realized. These findings will help clinicians identify patients with MDD and differentiate them from non-depressed patients with medical illness. PMID- 8227890 TI - Epidemic keratoconjunctivitis in a chronic care facility: risk factors and measures for control. AB - OBJECTIVE: To study patterns of transmission of epidemic keratoconjunctivitis (EKC) in a chronic care facility and to assess control measures and prevent future outbreaks in this setting. DESIGN: A retrospective cohort study. SETTING: A 120-bed, four-unit, skilled nursing facility. PATIENTS: Residents and employees of the above facility. INTERVENTIONS: Increased frequency of cleaning; use of bleach disinfectant; universal precautions in handling eye secretions from residents with conjunctivitis; cohorting residents by unit; suspension of new admissions; closure of common gathering areas. MEASUREMENTS: Resident demographics; possible risk factors for infection among residents (including mobility, underlying illness, medications, involvement in social activity, level of confusion) and among employees (including co-morbid illnesses and eye conditions, exposures to persons with conjunctivitis, visits to eye care specialists, use of contact lenses or glasses); testing of conjunctival specimens from symptomatic persons for viral and bacterial agents. RESULTS: Of 95 residents on three chronic care units, 47 (attack rate 49%) had onset of eye symptoms consistent with EKC between September 14 and December 7, 1990. Thirty-eight (81%) of these had onset following the onset of symptoms in a resident with dementia who, despite habitual eye-rubbing and wandering into other residents' rooms, was not isolated or restricted in any way. Attack rates were higher (though not statistically significant) among more mobile residents (60% for ambulatory residents) and among those considered by staff to be confused (56%). Rapid antigen detection and culture confirmed adenovirus type 37 as the etiologic agent. CONCLUSIONS: Transmission of infection with adenovirus type 37 was successfully interrupted following strict infection control, suspension of new admissions, cohorting of residents by unit, and change to a disinfectant that inactivates adenovirus. Recognition of conjunctivitis as an appropriate reason for restricting movement of an infected resident may have prevented extensive viral transmission in this outbreak. PMID- 8227891 TI - Reliability of sampling urine from disposable diapers in elderly incontinent women. AB - OBJECTIVE: To assess the reliability for biochemical and microbiological analysis of urine collection from disposable diapers in elderly women with severe urinary incontinence. DESIGN: Cross-sectional comparison of two methods of collection. PATIENTS AND MEASUREMENTS: Urine was sampled from 52 women inpatients (aged 68-98 years) in a geriatric hospital ward by pressing a diaper which the patient had worn for 3 hours. Just after this collection, another sample was obtained by retrograde catheterization. Both samples were analyzed for sodium, potassium, chloride, proteins, urea, creatinine, calcium, and phosphate and for cell counts, the presence of bacteria, and bacteria culture. RESULTS: For all the biochemical parameters, the urinary concentrations obtained by the two methods were strongly and significantly correlated. The following differences (mean and range, in mmol/L) were found between the values obtained by the two methods: sodium: 6.05 ( 26 to 27), potassium: (-16 to 14), chloride: -1.13 (-24 to 23), urea: 6.85 (-33 to 37), creatinine: 0.24 (-0.95 to 1.45), calcium: -0.22 (-1.27 to 1.70), and phosphate: 2.17 (-2.5 to 13.4). For diagnosis of urinary tract infection, agreement between the two methods was good (kappa = 0.84), and bacteriological agreement was obtained in 25 out of 28 cases (89%). However, for diagnosis of microscopic hematuria, agreement was poor (kappa = 0.50), probably due to the overestimation of the true urinary red cell count in the samples collected by catheterization. CONCLUSION: For routine microbiological and biochemical analysis, urine extraction from disposable diapers is a simple and reasonably reliable method of sampling urine from elderly women with severe incontinence. PMID- 8227892 TI - Fever of unknown origin in elderly patients. AB - OBJECTIVE: To describe the spectrum of diseases that may give rise to fever of unknown origin in elderly patients and to delineate the diagnostic approach in these patients. DESIGN: Subgroup analysis of a prospectively collected case series followed more than 2 years. SETTING: General Internal Medicine Service based at University hospital, Leuven, Belgium. PATIENTS: Forty-seven consecutive patients, older than 65 years, meeting the classic criteria of fever of unknown origin. MEASUREMENTS: The final diagnosis established and the clinical value of diagnostic procedures. RESULTS: Infections, tumors and multisystem diseases (encompassing rheumatic diseases, connective tissue disorders, vasculitis including temporal arteritis, polymyalgia rheumatica, and sarcoidosis) were found in 12 (25%), six (12%) and 15 patients (31%), respectively. Drug-related fever was the cause in three patients (6%), miscellaneous conditions were found in five patients (10%), and six patients (12%) remained undiagnosed. Microbiologic investigations were diagnostic in eight cases (16%), serologic tests yielded one diagnosis, immunologic investigations had a diagnostic value in four cases, standard X-rays yielded a diagnostic contribution in 10 cases, ultrasonography and computed tomography were diagnostic in 11 cases, Gallium scintigraphy had a diagnostic contribution in 17 cases, and biopsies yielded the final diagnosis in 18 cases. CONCLUSIONS: Multisystem diseases emerged as the most frequent cause of fever of unknown origin in the elderly, and temporal arteritis was the most frequent specific diagnosis. Infections, particularly tuberculosis, remain an important group. The percentage of tumors was higher in our elderly patients than in the younger ones but still clearly lower than in other recent series of FUO in adults. The number of undiagnosed cases was significantly lower in elderly patients than in younger individuals (P < or = 0.01). The investigation of elderly patients with FUO should encompass routine temporal artery biopsy and extensive search for tuberculosis if the classic tests such as blood count, chemistry, urinalysis, cultures, chest X-rays, and abdominal ultrasonography do not yield any clue. Gallium scintigraphy should be considered as the next step and not as a last-resort procedure. PMID- 8227893 TI - The relationship between age, gender and cognitive performance in the very old: the effect of selective survival. AB - OBJECTIVE: To determine the prevalence of cognitive disability as a function of advanced age and gender in elderly nursing home and community-dwelling populations. Since cognitive dysfunction is associated with increased mortality, we hypothesized that selective survival results in a decreased prevalence of cognitive disability in the oldest old. DESIGN: Cohort study. An analysis of 6 month longitudinal data obtained from a national probability sample of older persons in 260 nursing homes (n = 1951) and 2-year-longitudinal data obtained from a sample of community-dwelling older persons (n = 2947). MEASURES: In the nursing home sample, the primary outcome measure was cognitive performance score. In the community sample, cognitive performance was determined using the results of three orientation questions and assessment of decision-making ability. Cognitive performance and subsequent survival, controlling for various disease states and demographic factors, were examined in three age cohorts of men and women (ages 65-79, 80-89, 90-99). RESULTS: In the nursing home sample, the cognitive performance of very old men (> or = 90 years) was better than that of younger men (aged 80-89 years, P < 0.05) and very old women (age > or = 90 years, P = 0.001). Among 80-89-year-olds with poor cognitive performance, the 6-month mortality rate was higher in men than in women (38% vs 19%, P = 0.001). However, the mortality rates of men and women with good cognitive performance were not statistically different in any age group. Proportional-hazards regression analysis demonstrated that poor cognitive performance remained a powerful predictor of death among men aged 80-89 years with a relative risk of 2.7 (95% Cl, 1.19-3.17; P = 0.0006) after controlling for covariates. Results from the community sample lent support to our findings: within each age group, mortality rates for men and women with intact cognitive performance were not statistically different. However, in the two older age groups, the mortality rates of subjects with impaired cognitive performance were significantly greater for men than for women (P < 0.01 for both age groups). CONCLUSIONS: Decreased cognitive performance is significantly associated with mortality among elderly men. Survival by men who have relatively intact cognitive function results in a population of oldest men, those aged 90-99 years, with cognitive performance scores better than younger men or similarly-aged women. The same selective survival phenomenon was not observed among women. Thus, there may be less cognitive disability among very old men than previously expected. PMID- 8227894 TI - The effects of age and diabetes mellitus on nerve function. AB - OBJECTIVE: This project evaluated the association of age and vibratory thresholds (assessment modality of large sensory nerve fibers) in subjects with and without diabetes mellitus. DESIGN: Cross-sectional study. SETTING: Medical Research Institute of Delaware. PARTICIPANTS: Individuals with non-insulin-dependent diabetes mellitus and non-diabetic control subjects. MEASUREMENTS: Vibratory thresholds were examined in four age groups (ie, < 45 yrs, 45-54 yrs, 55-64 yrs, > or = 65 yrs). The independent association of age, duration of diabetes, height, gender, glycemic control, and smoking history were analyzed in terms of their relationship to vibratory thresholds. MAIN RESULTS: Vibratory thresholds increased with age for both control and diabetic subjects. Comparing controls with diabetic subjects in the same age categorizes revealed significant differences for vibratory thresholds only in the > or = 65 year old age group. Modeling with vibratory thresholds as the dependent variable showed that age and male gender were independently associated with vibratory thresholds for the controls and explained the majority of the variability (R2 = 0.79). Age, duration of diabetes, and height were independently associated with vibratory thresholds for the diabetic subjects but explained much less of the variability (R2 = 0.39). CONCLUSIONS: The results suggest an acceleration of the natural aging process for large sensory nerve fiber function in diabetic subjects. Thus, young diabetic subjects may be at a risk of lower extremity complications as a result of injuries similar to that older non-diabetic individuals. PMID- 8227895 TI - Joint impairment and ambulation in the elderly. AB - OBJECTIVE: To test the impact of joint impairment on ambulation in the elderly, using a multivariate model. DESIGN: Cross-sectional observational study (baseline data from an ongoing longitudinal study). SUBJECTS: Five hundred thirty-two persons over age 60, including continuing care retirement community (CCRC) (n = 222), homebound (n = 63), and ambulatory (n = 247) respondents. Mean age at assessment = 76.6 (SD = 6.6). MEASUREMENT: Independent variables included sociodemographics, physician measures of lower joint impairment, an index of cormorbidities derived from physical examination or chart abstract, self-assessed arthritis pain, depression, and anxiety. The dependent variable, ambulation, was measured as walk-rate, based on the time required to walk 50 feet. MAIN RESULTS: For the total sample, 61% of the subjects were impaired in more than one lower joint group, with almost 50% of the homebound impaired in more than three joint groups. Demographics (particularly age and education) explained much of the variance in walk-rate. After controlling for demographics and membership in the groups purposely sampled (CCRC, homebound, ambulatory), lower joint impairment accounted for an additional 7% of the variance in walk-rate. Total amount of variance explained by the model was 56% (Adjusted R2 = .56). A 3-point change in lower joint impairment score, equivalent to the maximum impairment score for a single joint group, is associated with a 4-second change in the mean time required to walk 50 feet. The knee and lower spine joints contributed most to the impact of the lower joint impairment measure. CONCLUSIONS: Among the elderly, age and education are strong predictors of performance on a walk-rate test. Independent of demographics and non-musculoskeletal conditions, joint impairment is associated with diminished walking ability in this population. PMID- 8227896 TI - Malnutrition in institutionalized elderly: how and why? AB - OBJECTIVE: To determine the prevalence of undernutrition and overnutrition in long-term care elderly patients and the functional, behavioral, environmental, nutritional, and medical variables associated with this prevalence. DESIGN: Cross sectional, observational. SETTING: Long-term care hospital in Canada. SUBJECTS: Two hundred elderly patients (n = 166 male), average age 78.5 years. MEASUREMENTS: Assessment of nutritional status and presence of specific behavioral, medical, environmental, and functional characteristics known to impact on nutritional status. Nutritional status was determined by weight, % weight loss, BMI, skinfolds, arm circumference, area measurements, and % body fat. Multiple regression analyses were performed to identify the factors associated specifically with undernutrition and overnutrition in this population. RESULTS: Severe undernutrition was present in 18% (n = 36) and severe overnutrition in 10% (n = 20). Mild/moderate undernutrition was present in 27.5% (n = 55) and mild/moderate overnutrition in 18% (n = 36). Overnutrition was positively associated with primary diagnosis and number of medications and negatively associated with poor appetite, number of feeding impairments, protein intake, and mental state. Undernutrition was positively associated with dysphagia, slow eating, low protein intake, poor appetite, presence of a feeding tube, and age and negatively associated with primary diagnosis. CONCLUSIONS: Undernutrition exists at a level that is high (45.5%) but not unusual for this type of institutional setting. Behavioral, environmental, and disease-related factors greatly influence nutritional status. Undernutrition appears to be affected by nutritional factors more than overnutrition. Efforts should be directed toward influencing some of these factors to decrease undernutrition in the institutionalized elderly. PMID- 8227897 TI - L-deprenyl in the treatment of mild dementia of the Alzheimer type: results of a 15-month trial. AB - OBJECTIVE: To examine the cognitive and behavioral effects of L-deprenyl in persons with mild dementia of the Alzheimer type (DAT) over a 15-month period. DESIGN: A 15-month randomized, double-blind, placebo-controlled trial using a parallel-group design. PARTICIPANTS: 39 subjects with mild DAT (CDR 1) selected using NINCDS-ADRDA criteria. OUTCOME MEASURES: A battery of neuropsychological tests and clinical rating scales. RESULTS: The placebo and L-deprenyl subjects were similar at baseline on the clinical and neuropsychological tests. The total score on the Brief Psychiatric Rating Scale (BPRS) was significantly less after 15 months in subjects taking L-deprenyl. The only individual item of the BPRS which differed between groups was disorientation. There was no evidence of a L deprenyl effect on any other clinical or neuropsychological measure after 2, 8, or 15 months. CONCLUSION: L-deprenyl did appear to have a slight effect on a single measure of psychopathology, the BPRS. It did not have a measurable impact on any other measure of behavior or cognitive function over a 15-month period in this group of subjects with mild DAT and did not appear to slow the progression of the disease. PMID- 8227898 TI - Type of fall and risk of hip and wrist fractures: the study of osteoporotic fractures. The Study of Osteoporotic Fractures Research Group. AB - OBJECTIVE: To determine the causes of hip or wrist fractures. DESIGN: Case control analysis nested in a prospective cohort study. SETTING: Four clinical centers in Baltimore, Maryland, Minneapolis, Minnesota, Portland, Oregon, and Monessen, Pennsylvania. PARTICIPANTS: Non-black women age 65 and older living in the community. MEASUREMENTS: We measured bone density, arm strength, and anthropometric characteristics at a baseline examination. Those who subsequently suffered hip (n = 130) or wrist (n = 294) fractures as a result of a fall and a consecutive sample of those who fell without a fracture (n = 467) were interviewed about their falls. RESULTS: In multivariate analyses, those who suffered hip fractures were more likely to have fallen sideways or straight down (odds ratio 3.3; 95% confidence interval 2.0 to 5.6) and to have landed on or near the hip (32.5; 9.9, 107.1) than women who fell without a fracture. Among women who fell on the hip, those with hip fractures were taller (1.5; 1.2, 2.2 per SD increase), less likely to have landed on a hand (0.3; 0.1 to 0.6) or to break the fall by grabbing or hitting an object (0.4; 0.2, 0.9), had weaker triceps (1.7; 1.2, 2.5 per SD decrease), and were more likely to land on a hard surface (2.8; 1.4, 5.5) than those without fractures. Women with wrist fractures were more likely to have fallen backward (2.2; 1.3, 3.8) and to have landed on a hand (20.4; 11.5, 36.0) than those who fell without a fracture. Among women who fell on the hand, those with wrist fractures were taller (1.4; 1.1, 1.8 per SD increase) and less likely to break the fall by grabbing or hitting an object (0.4; 0.2, 0.7). Among women who fell on their hip or hand, the risk of fracturing that site more than doubled for each standard deviation decrease in bone density at the site of fracture. CONCLUSIONS: The nature of the fall determines the type of fracture, while bone density and factors that increase or attenuate the force of impact of the fall determine whether a fracture will occur when a faller lands on a particular bone. These findings have important implications for prevention of fractures in older women. PMID- 8227899 TI - Clock completion: an objective screening test for dementia. AB - OBJECTIVE: To develop a simple, readily administered and scored screening test for dementia utilizing the clock-drawing task. DESIGN: Retrospective analysis of clock-drawing errors and prospective validations. SETTING: Hospital-based outpatient geriatric assessment clinic, rehabilitation service, apartment building for older adults, and long-term care facility. PARTICIPANTS: Convenience sample of patients attending the geriatric assessment clinic, patients on the rehabilitation service, or residents of the above sites. MEASUREMENTS: Sensitivity and specificity of a clock-scoring system in identifying patients with dementia and the comparison of this system with the Short Blessed Test (SBT) in the diagnosis of dementia and in the prospective validation of the test. RESULTS: Of the 10 clock-drawing errors evaluated, placement of digits in a pre drawn circle had the greatest sensitivity and specificity in distinguishing patients with irreversible dementia from patients with other disorders who did not meet NINCDS-ADRDA criteria for probable dementia. The derived scoring system had a sensitivity of 87% and a specificity of 82%, compared with a sensitivity of 82% and a specificity of 88% for the SBT in identifying dementia. Test-retest reliability for the distinction between demented and non-demented was 82%, with a Kappa of 0.63 for the clock completion, and 82%, with a Kappa of 0.62 for the SBT. Inter-rater reliability for clock completion was 0.90 to 0.93. CONCLUSION: A simple, completely objective scoring system for a clock completion test has been developed which involves only the number of digits placed in the fourth quadrant of a pre-drawn circle. This readily administered test is as effective in screening for dementia as the longer six-item SBT. PMID- 8227900 TI - Comparison of cyclic guanosine monophosphate response to infusion of atrial natriuretic peptide in young and elderly subjects. AB - OBJECTIVE: To compare the sensitivity to atrial natriuretic peptide (ANP) in young and elderly subjects. DESIGN: Double-blind, random-order, placebo controlled study using two different rates of ANP infusion. SUBJECTS: Eight healthy young (23 +/- 3 years, mean +/- SD) and eight healthy elderly (76 +/- 4 years) subjects recruited from the general population. MEASUREMENT: Plasma cyclic guanosine monophosphate (cGMP, the second messenger of ANP) levels reflecting the sensitivity of biologically active ANP receptors. RESULTS: Baseline ANP levels were higher and rose to a greater extent in the elderly both during low- (0.25 microgram/min) and high-dose (2.0 micrograms/min) infusion of ANP. Plasma levels of cGMP rose in proportion to the increase in ANP. When the rise in cGMP levels was related to the increase in ANP levels (expressed as nmol/L per 100 pg/mL ANP increase), no difference was found between young and elderly subjects. CONCLUSION: The ability of elderly subjects to increase cGMP in response to exogenous ANP is similar to that of young subjects. This may reflect a similar sensitivity of the target organs towards ANP. However, because of lower clearance in the elderly, a similar dose of ANP results in a greater elevation of ANP and cGMP levels and hence induces a greater effect. PMID- 8227901 TI - Evidence for a genetic basis for the model system of cellular senescence. PMID- 8227902 TI - Total quality management: administrative and clinical applications in nursing homes. AB - OBJECTIVE: Describe the application of the Total Quality Management (TQM) model to the unique work force, resident population, and regulatory issues that characterize long term care settings. DESIGN: The key differences between TQM and current management and training practices in nursing homes are described. A specific data-based example is provided of a successful TQM application to health care involving clinical work processes related to incontinence care. CONCLUSION: Significant organizational and clinical obstacles must be overcome if TQM is to improve the quality of life and satisfaction of nursing home residents and their families as it has improved the efficiency and product quality in hospital and non-health-care settings. PMID- 8227903 TI - The relationship between aging and disease: geriatric ideology and myths of senility. AB - An ongoing debate in gerontology concerns the relationship between aging and disease. Goodwin, as part of a discussion on the importance of geriatric ideology, argues that age-related disorders like senile dementia of the Alzheimer's type (SDAT) should be regarded as an aging process, while others suggest it is more appropriate to consider it as a disease. This paper comments on the arguments presented by Goodwin and develops these by providing a review of the underlying assumptions concerning aging and disease. The policy relevance and implications for future research are also outlined. PMID- 8227904 TI - The aging-disease dichotomy is alive, but is it well? PMID- 8227905 TI - Unaffordable cost of pharmaceuticals: a cause for suboptimal compliance. PMID- 8227906 TI - Effect of aging on body fat. PMID- 8227907 TI - Videofluoroscopic swallowing studies. PMID- 8227908 TI - CPR in nursing homes. PMID- 8227909 TI - Metabolic and behavioral covariates of high-density lipoprotein cholesterol and triglyceride concentrations in postmenopausal women. AB - OBJECTIVE: To determine predictors of high-density lipoprotein cholesterol (HDL) and triglyceride (TG) concentrations in postmenopausal women. DESIGN: Cross sectional study. SETTING: Clinical research facility. PARTICIPANTS: One hundred twenty-seven healthy, relatively sedentary, postmenopausal women not on estrogen replacement, mean age 57 years. MEASUREMENTS: Alcohol intake, cigarette smoking, aerobic fitness (VO2max), body mass index (BMI), percent body fat, waist-hip ratio, lipids and lipoproteins, fasting plasma glucose (FPG), and insulin (FPI) concentrations. RESULTS: In univariate analyses, HDL was significantly (P < 0.05) inversely related to BMI, waist-hip ratio, smoking, FPG, and FPI, and directly related to VO2max and alcohol intake. Triglycerides were related directly to BMI, waist-hip ratio, percent body fat, FPG, and FPI, and inversely to VO2max. In stepwise multiple regressions, BMI, waist-hip ratio, alcohol, smoking, and FPG were significantly associated with HDL (R2 for the model = 0.43). Addition of TG to these models reduced relations of BMI and waist-hip ratio, but not the other variables, to insignificance. For triglycerides, waist-hip ratio, alcohol, smoking, FPG, and FPI were significant predictors (R2 = 0.33). VO2max and percent body fat did not contribute to any model. CONCLUSIONS: Obesity, abdominal obesity, smoking, alcohol intake, and measures of carbohydrate metabolism predict HDL and triglyceride concentrations in postmenopausal women. PMID- 8227910 TI - Memory complaints of elderly people in a population survey: variation according to dementia stage and depression. AB - OBJECTIVE: To study the relationship between subjective memory complaints, cognitive functioning, and clinical diagnosis of dementia. DESIGN: Cross sectional, total-population survey. MEASUREMENTS: A two-phase study was done. All 2368 were screened by Mini-Mental State Examination (MMSE). Then the 314 who screened positive and a sample of 354 who screened negative were examined clinically (by examining the subject and interviewing an informant) for DMS-III-R criteria for dementia, Clinical Dementia Rating (CDR) scale for severity of dementia and the presence of memory complaints or depressed mood. SETTING: An area in Stockholm, Sweden. PARTICIPANTS: All inhabitants of the area (2368) born in 1912 or before, living at home or in institutions. RESULTS: A greater percentage of informants reported marked memory impairment than did the subjects themselves. Neither the informants' nor the subject' reports differed according to the type of dementia, but they differed clearly according to the stage of dementia. Informants reported memory impairment more frequently as dementia severity increased, while the subjects' complaints of marked memory impairment were most common in mild-moderate dementia. Furthermore, non-demented subjects with marked complaints performed more poorly on the MMSE. Subjects with depressed mood were also more likely to complain about their memory, both in the demented and non-demented groups. CONCLUSION: Our data show the validity of the informants' reports about the subjects' memory deficits. Moreover, the study confirmed the clinical impression that the patient's own complaints should not be ignored, as subjects in the mild stage of dementia often have some insight into their own memory deficit. However, depressed individuals may underestimate their own memory, and individuals with moderate-severe dementia may overestimate theirs. PMID- 8227911 TI - Serum erythropoietin levels in elderly inpatients with anemia of chronic disorders and iron deficiency anemia. AB - OBJECTIVE: To analyze the relationship between serum erythropoietin levels and hemoglobin levels in elderly patients with anemia of chronic disorders related to cancer or acute infection when compared with anemic patients with iron deficiency. DESIGN: Prospective survey with comparison groups. SETTING: Tertiary care center. PATIENTS: An elderly group aged 70 and above (mean 84, range 70-96) was divided into subgroups of 45 with anemia of chronic disorders (23 with cancer and 22 with acute infection), 24 with iron-deficiency anemia, and 27 with no anemia. Thirty non-anemic younger adults were also studied. MEASUREMENTS: Serum erythropoietin (radioimmunoassay), complete blood count, serum iron, B12, folate and ferritin, liver and kidney function tests, blood gas analyses, and bacteriological and radiological tests. RESULTS: The serum erythropoietin levels were significantly lower in the elderly non-anemic hospitalized group than in the healthy younger group. A significant negative relationship between the log serum erythropoietin and hemoglobin levels was found in patients with iron deficiency, but not in the other groups. For any given hemoglobin level, the response of erythropoietin was significantly higher in anemic patients with iron deficiency when compared with the neoplastic and infectious group. CONCLUSION: Erythropoietin response to anemia is blunted in elderly patients with anemia of chronic disorders related to cancer or acute infection. Erythropoietin level is lower in non-anemic elderly inpatients than in healthy younger persons. PMID- 8227912 TI - Verbal learning and/or memory improves with glycemic control in older subjects with non-insulin-dependent diabetes mellitus. AB - OBJECTIVE: To determine whether cognitive function improves with improved glycemic control in older subjects with non-insulin-dependent diabetes (NIDDM). We hypothesized that with improved glycemic control: 1) learning and memory, 2) attention, and 3) complex perceptual-motor function would improve, but that 4) simple perceptual-motor function would not. DESIGN: Non-randomized control trial. SETTING: Aging Study Unit, Department of Veterans Affairs Medical Center. SUBJECTS: Thirty subjects with NIDDM; 17 on oral hypoglycemic agents; 13 untreated at study entry. Thirteen normal controls. INTERVENTION: Subjects on oral hypoglycemic agents were taken off medications. After 1 month, they and previously untreated subjects began treatment with glipizide. Dose was titrated up weekly until fasting plasma glucose was less than 7.8 mmol/L or maximal dose (40 mg/day). Controls received no medication. MEASUREMENTS: Fasting plasma glucose (FPG), glycated hemoglobin, and measures of cognitive function in four general categories: 1) learning and memory, 2) ability to sustain attention, 3) complex perceptual-motor function, and 4) simple perceptual-motor function. All were evaluated in subjects with NIDDM at baseline (T1), after 1-month washout (T2), and after 2 (T3) and 4 months (T4) of optimal glycemic control or maximal dose. Controls were evaluated at the same intervals. RESULTS: FPG and glycated hemoglobin rose in previously treated subjects from T1 to T2 (9.4 +/- SEM 0.4 to 14.7 +/- 0.7 mmol/L and 10.9 +/- 0.7% to 12.2 +/- 0.6%, respectively) but were unchanged in previously untreated subjects (11.3 +/- 0.6 to 11.8 +/- 0.9 mmol/L and 10.9 +/- 0.7% to 11.7 +/- 0.7%). With glipizide treatment, there was a decrease in FPG level at T3 (9.4 +/- 0.5 mmol/L in previously treated, 6.9 +/- 0.4 mmol/L in previously untreated), which persisted at T4. Glycated hemoglobin fell similarly. FPG and glycated hemoglobin were unchanged in controls. As hypothesized, learning and memory improved over time with treatment in both groups of subjects but was unchanged in controls (P < 0.05). Detailed analysis indicated that the improvement occurred primarily in the learning of verbal material. Contrary to hypothesis, attention and complex perceptual-motor function did not show improvement. As expected, simple perceptual-motor function did not show any improvement with treatment. CONCLUSIONS: The results are consistent with previous findings that poor glycemic control in older subjects with NIDDM is associated with decreased cognitive functioning, and suggest that verbal learning and memory may improve with improved glycemic control. PMID- 8227913 TI - Depression and smoking cessation in older adults: a longitudinal study. AB - OBJECTIVE: To explore the relationship of smoking cessation and depression. DESIGN: Cohort study with 3 years of follow-up. SETTING: North Carolina community of the Established Populations for the Epidemiologic Studies of the Elderly. PARTICIPANTS: Stratified cluster sample of adults age 65 years and older, with an oversampling of African Americans. MAIN MEASURES: Depressive symptoms using a modified Center for Epidemiologic Studies Depression (CES-D) scale; smoking cessation using change in self-reported smoking. RESULTS: Current smokers had the highest prevalence of clinically significant CES-D scores (11.2%), followed by never smokers (9.6%) and former smokers (7.1%). After 3 years, 128 (25%) of 511 baseline current smokers had quit. Among women with a clinically significant CES D score, 55% quit smoking, compared with only 25% among those with a normal score (P < 0.05). Depressive symptoms were significantly associated with nearly fourfold increased odds of smoking cessation among women (relative odds [RO] 3.7, 95% confidence interval [CI] 1.2, 11.0; P < 0.05), but not among men (RO 0.6, 95% CI 0.2, 2.0, not significant), after adjusting for potential confounding. CONCLUSIONS: Depressive symptoms may directly increase the likelihood of smoking cessation among older women. When predicting smoking cessation, depression and gender should be considered in combination since to consider them separately may be misleading. This challenges prior reports that depressed smokers are less likely to quit smoking than nondepressed smokers. PMID- 8227914 TI - Comparison of clinical indicators in two nursing homes. AB - OBJECTIVE: Pressure ulcer prevalences in 30 VA nursing homes in 1986 ranged from 0% to 15%. The institutions with lowest ("A") and highest ("B") prevalence were selected for further examination. DESIGN: Analysis of nursing home files for five study periods, each lasting 6 months. SETTING: A and B were 60-bed rural and 280 bed urban facilities, respectively. MEASUREMENTS: Eleven outcome indicators were calculated for each study period: prevalences and incidences of pressure ulcer, aggressive behavior and disruptive behavior, 6-month declines in each of the four activities of daily living (ADLs), and prevalence of underweight. RESULTS: Populations in A and B were similar with regard to age, sex, length of stay, degree of dependency, and level of nursing care. All indicators for the first study period were more favorable in A than in B. In addition, underweight (body mass index < 22 kg/M2) was significantly less prevalent in A than in B. The differences between the two institutions in the indicators were persistent over the five study periods from 1988 to 1991. CONCLUSIONS: The populations of A and B were similar in the available measures of severity of illness. Nevertheless, the residents in nursing home A were significantly less likely to experience adverse outcomes than were the residents in nursing home B. The virtual absence of pressure ulcers, physical aggression, and verbal disruption in nursing home A, despite the presence of many immobile and demented residents, suggested that these complications can mostly be prevented. PMID- 8227915 TI - Adverse events related to drugs and drug withdrawal in nursing home residents. AB - OBJECTIVE: To (1) develop and standardize explicit criteria to link clinical adverse events to drug withdrawal, (2) determine the incidence and severity of Adverse Drug Events (ADEs) and Adverse Drug Withdrawal Events (ADWEs) in a nursing home population, and (3) establish the contribution of demographic, clinical, and functional characteristics to ADEs and ADWEs. DESIGN: Retrospective record review of an admission cohort. SETTING AND SUBJECTS: Consecutive admissions of residents of an academic Veterans Affairs nursing home with available records and lengths of stay > 30 days (n = 175). Subjects were 96% men, aged 70 +/- 12 years, and took 7.0 +/- 3.4 medications. METHODS: We applied standardized algorithms to determine incidence, probability, and severity of ADEs and ADWEs. Multiple regression techniques were used to identify factors associated with frequency and risk of events. RESULTS: Ninety five residents experienced 201 ADEs. Twelve required hospitalization or prolonged hospitalization, and one resident died. Sixty two persons had 94 ADWEs. None were associated with death and one with hospitalization. The four most commonly prescribed drug classes accounted for 72% of ADEs and 80% of ADWEs. Results of multivariate analyses showed common risk factors for both ADEs and ADWEs: number of diagnoses, number of medications, and hospitalization during the nursing home stay. CONCLUSIONS: ADEs and ADWEs were common in nursing home residents in this Veteran's Affairs setting. Explicit criteria developed and applied in this study should be applied prospectively in other settings, both to further define risk of drug discontinuation and to assist in development of specific drug discontinuation guidelines. PMID- 8227916 TI - Over 80 years and no evidence of coronary heart disease: characteristics of a survivor group. AB - OBJECTIVE: To determine factors that characterize those 80 years and over free of clinical coronary heart disease (CHD). DESIGN: Cross-sectional intergroup comparison. SETTING: Rural New Zealand town. SUBJECTS: All those 70 years and over registered with the sole health center. Seven hundred eighty-two subjects were seen, a 91.4% response rate. Subjects were divided by World Health Organization cardiovascular questionnaire criteria, history, examination, and coded 12-lead electrocardiogram into four groups: 1) those 70-79 years of age with evidence of CHD; 2) those 80 years and older with evidence of CHD; 3) those 70-79 years of age free of CHD; and 4) those 80 years and older free of clinical CHD. MEASUREMENTS: Mean values and distributions of cholesterol, high-density lipoprotein (HDL) cholesterol, apolipoproteins (apos), triglycerides, insulin, glucose, anthropometric measurements, and psychosocial variables. RESULTS: In the initial analysis of variance there was a significant difference in the mean values among the four groups for body mass index (BMI), HDL cholesterol, apo A1, and triglycerides. Using the Duncan procedure, the mean value of HDL cholesterol in the group 80 years and older with no evidence of clinical CHD was significantly higher than in each of the other groups. The group 80 years and older without clinical CHD had a significantly higher mean value for apo A1 and significantly lower mean values for triglycerides and BMI than the group 70-79 years old with CHD (p < 0.05). Three-factor analysis of variance showed those free of clinical CHD had significantly higher values of HDL cholesterol (P = 0.024) and apo A1 (P = 0.022), and lower values of BMI (P = 0.017) and triglycerides (P = 0.018) compared with those who had CHD after controlling for age and sex. Those 80 years and older had lower values of BMI (P = 0.001) and triglycerides (P = 0.018) than those 70-79 years old after controlling for CHD and sex. The group who were 80 years and older and free of clinical CHD had a significantly narrower distribution of values for insulin (P < 0.001), glucose (P = 0.001), diastolic blood pressure (P = 0.001), BMI (P = 0.001), and waist-hip ratio (P = 0.002) when compared with those 80 years and older with CHD; a significantly narrower distribution of values for insulin (P = 0.024), glucose (P = 0.003), and BMI (P = 0.002) when compared with those 79-79 years old with CHD and a significantly narrower distribution of glucose (P = 0.002) and BMI (P = 0.014) when compared with those 70-79 years old and free of clinical CHD. CONCLUSION: Those 80 years and older who were free of clinical CHD were characterized by differences in lipid and anthropometric values, particularly higher HDL cholesterol and apo A1 levels. Results were also consistent with this survivor group maintaining tighter homeostatic control over a number of variables. PMID- 8227917 TI - Physical activity and serum lipids and lipoproteins in elderly women. AB - OBJECTIVE: The relationship between cardiorespiratory exercise and serum lipid and lipoprotein levels was studied in elderly women. DESIGN: Randomized controlled experimental design with a follow up of 12 weeks; cross-sectional comparison at baseline. SETTING: Community-living elders in university exercise facilities. PARTICIPANTS: Thirty-two apparently healthy, sedentary elderly Caucasian women, 67 to 85 years of age. Ten highly conditioned elderly women, 65 to 84 years of age, who were active in endurance competitions and had been training for 11.2 +/- 1.2 years, were recruited at baseline for cross-sectional comparisons. INTERVENTIONS: Sedentary subjects were randomized to either a walking or calisthenic group. Intervention groups exercised 30 to 40 minutes, 5 days a week for 12 weeks, with the walking group training at 60% heart rate reserve and the calisthenic group engaging in mild range-of-motion and flexibility movements that kept their heart rates close to resting levels. MEASUREMENTS: Serum lipids and lipoproteins, maximal aerobic capacity (VO2 max), four skinfolds, and dietary intake at baseline and after 5 and 12 weeks. RESULTS: When the highly conditioned group and combined group of sedentary subjects were compared at baseline, serum high-density lipoprotein cholesterol (HDL-C; 1.61 +/- 0.14 vs 1.27 +/- 0.05 mmol/L, respectively; P = 0.048) and triglycerides (1.29 +/ 0.15 vs 2.00 +/- 0.15, respectively; P = 0.002), but not total serum cholesterol (5.72 +/- 0.36 vs 5.72 +/- 0.19 mmol/L, respectively) and low-density lipoprotein cholesterol (LDL-C; 3.62 +/- 0.36 vs 3.72 +/- 0.18 mmol/L, respectively), were significantly different. Twelve weeks of moderate cardiorespiratory exercise improved the VO2max of the sedentary subjects 12.6% but did not result in any change in body weight, energy intake, dietary quality, or any of the serum lipids or lipoproteins. CONCLUSION: Highly conditioned and lean elderly women, when compared with their sedentary counterparts, had higher HDL-C and lower triglycerides, but similar total serum cholesterol and LDL-C values. However, twelve weeks of moderate cardiorespiratory exercise were not associated with an improvement in serum lipid or lipoprotein profiles in previously sedentary elderly women. PMID- 8227918 TI - The Yale Geriatric Care Program: a model of care to prevent functional decline in hospitalized elderly patients. AB - OBJECTIVE: To describe the structure and implementation of the Yale Geriatric Care Program, an innovative, nursing-centered model for developing geriatric nursing expertise throughout an acute-care hospital. DESIGN: Descriptive study of an intervention in a prospective cohort of patients. SETTING: University teaching hospital. PATIENTS: Two hundred forty four patients aged 70 years and older on four non-intensive care intervention units during the study period (July 9, 1990 to July 31, 1991). INTERVENTION: The Geriatric Care Program involved an integrated model of primary nurses, specially trained unit-based geriatric resource nurses, gerontological nurse specialists, and geriatric physicians. The intervention included surveillance and identification of frail older patients, unit-based geriatric educational programs for all nurses, special education and support for the geriatric resource nurses, and twice-weekly rounds of the Geriatric Care Team. RESULTS: The Geriatric Care Program has been successfully implemented on four units. The interventions ranged from general clarification of goals in 226 (92%) to specific recommendations for management of immobility in 100 (41%), bladder/bowel problems in 99 (41%), pressure ulcer treatment or prevention in 61 (25%), confusion evaluation or management in 62 (25%), and adjustment of medications in 43 (18%). Overall, 68% of the specific recommendations were documented to have been implemented. Barriers to implementation of the program have included initial difficulties with recruitment and retainment of geriatric resource nurses (due to high nursing turnover and the increased time commitment required), breakdown in communication and carryover of recommendations between nursing shifts, and obstacles to communication between the nursing and medical staff. CONCLUSIONS: An innovative model of care, in which geriatric nursing was integrated as part of standard nursing care on selected medical and surgical units, has been designed and implemented. Evaluation of the effectiveness and costs of this intervention are currently underway. PMID- 8227919 TI - A controlled trial of a nursing-centered intervention in hospitalized elderly medical patients: the Yale Geriatric Care Program. AB - OBJECTIVE: To test the effectiveness of a nursing-centered intervention to prevent functional decline among hospitalized elderly medical patients. DESIGN: Prospective cohort study with stratified and matched cohort analyses. SETTING: General medicine wards of a university teaching hospital. PATIENTS: Two hundred sixteen patients aged > or = 70 years (85 intervention and 131 control patients). INTERVENTION: The intervention included identification and surveillance of frail older patients, twice-weekly rounds of the Geriatric Care Team, and a nursing centered educational program. MAIN OUTCOME MEASURE: Functional decline, defined as a net decline in five activities of daily living (ADLs). RESULTS: In stratified analyses, the intervention resulted in a beneficial effect with a relative risk of 0.82 (95% confidence interval [CI] 0.54 to 1.24) in patients (n = 106) with one of four geriatric target conditions at baseline (eg, delirium, functional impairment, incontinence, and pressure sores). The intervention had no effect in patients without target conditions at baseline (n = 110); thus, this subgroup was excluded from further analyses. When patients were matched on number of target conditions and risk for functional decline at baseline (n = 66), the intervention resulted in a significant beneficial effect, with a reduction in functional decline from 64% in controls to 41% in the intervention group, for a relative risk of 0.64 (95% CI, 0.43 to 0.96). The intervention group had significantly less decline in ADL score and in individual ADLs than control subjects. Specific interventions aimed at maximizing function, such as physical therapy, were received more often by intervention patients; however, the beneficial effects of the intervention were achieved without increasing per-day hospital costs. CONCLUSIONS: The intervention appears effective to decrease functional decline in targeted elderly hospitalized medical patients. PMID- 8227921 TI - Enteral feeding in the aged. PMID- 8227920 TI - Thyroid disease in the elderly. AB - Thyroid disease in the aged, both hypothyroidism and hyperthyroidism, may be subtle or may be present with no clinical symptoms and signs, and is therefore difficult to diagnose on the basis of clinical evaluation. The help of the laboratory is essential in making the diagnosis of disease of the thyroid. Therapeutic strategies are different in the aged than in the younger adult with thyroid disease. It is essential for geriatricians, and all clinicians who care for the elderly, to have a solid understanding of thyroid function and dysfunctions in this group of patients so that they diagnose diseases of the thyroid correctly and treat them appropriately. PMID- 8227922 TI - Vitamin D status in the elderly. PMID- 8227923 TI - Ticlopidine-induced prolonged cholestasis. PMID- 8227924 TI - Urinary retention in the elderly. PMID- 8227925 TI - Weighing geriatric patients. PMID- 8227926 TI - Rectal and gynecological pathologies in long-term care patients. PMID- 8227927 TI - Medicare reimbursement for physician nursing home visits. PMID- 8227929 TI - AMWA and health care reform. PMID- 8227928 TI - The politics of health care reform. PMID- 8227930 TI - Impact of the structure of medicine in Britain on the careers of women physicians. PMID- 8227931 TI - Women in medicine: toward a conceptual understanding of the potential for change. AB - In 1959 only 6% of Canadian medical school graduates were women, but by 1989 44% of graduating classes were female. Recent data indicate that women are more likely than men to work as general practitioners, in groups, in urban settings, and on salary; to work fewer hours per week and to see fewer patients. In this paper we address the ongoing discussion of the impact of women's progressive entry into medicine. We suggest that a shortcoming of the discussion thus far has been its preoccupation with a descriptive examination of women physicians' professional characteristics and practices; sufficient attention has yet to be paid to the development of a clearer conceptual understanding of the nature of gender differences and the potential for change they imply. Using data from quantitative and qualitative phases of a national study of Canadian physicians, we address the relationship between gender differences in practice and underlying attitudes and values and identify alternative hypotheses about the potential for change created as more women enter medicine. We suggest that this potential goes beyond professional demographics and workloads to the doctor-patient relationship, but that the extent of change will be mediated by the socializing impact of medical school and the extent to which women physicians are included in or excluded from positions of power within the profession. We suggest also, that to meaningfully understand and document change, greater attention must be given to arguments that women and men view the world in qualitatively different ways. PMID- 8227932 TI - Parenting and professionalism: competing and enriching commitments. AB - The goals, format, curriculum, and faculty of a first-year medical school elective focusing on balancing commitments to a medical career and family are described in order to encourage replication at other institutions. While the course is offered to both men and women, the issues discussed have particular relevance to women physicians, the majority of whom continue to carry the major responsibilities for family and household in addition to their medical careers. Topics include: the rewards of being a parent, parent-child attachment, the dual career family at different developmental stages, physicians' life-styles, family planning, child care and training options, specialty choice and practice alternatives. The eight sessions are presented through a combination of seminar, videotapes, readings, and interviews of physicians, their spouses and children. PMID- 8227933 TI - Rape report challenged. PMID- 8227934 TI - The big pupil. PMID- 8227935 TI - Pupillary dilation: a view from the trenches. PMID- 8227936 TI - VISION USA--optometry's national charity program. AB - BACKGROUND: There are an estimated 18 million uninsured working Americans and their families who are unable to afford the cost of eye care. These individuals often fall between the cracks of government and private health care assistance. Members of the American Optometric Association have developed the VISION USA program to provide free eye care to those who might otherwise not receive it. METHODS: Nearly 8,000 doctors of optometry nationwide have volunteered to provide free eye examinations in their offices for VISION USA patients. Patients are screened for eligibility and matched to the nearest available participating doctor using a computerized ZIP code matching system. Examinations are usually provided each year during March in observance of Save Your Vision Week. RESULTS: In the first three years of the program, services have been provided to over 115,000 individuals. For over 21 percent of the patients seen, this was their first eye examination ever. More than 86 percent of the patients had one or more vision problems. More than 70 percent were given new or updated prescriptions. CONCLUSION: The VISION USA program is helping to meet a significant unmet need for eye care which exists within a segment of the American population. The program is an outstanding example of the willingness of doctors of optometry to give back to their communities, and to those in need, some of what they have gained from the practice of their profession. PMID- 8227937 TI - Efficacy of a pediatric cycloplegic administered as a spray. AB - BACKGROUND: The topical administration of eyedrops is often an unpleasant procedure in the pediatric population. We sought to learn whether a combination mydriatic-cycloplegic solution would be an effective cycloplegic if delivered as a spray. METHODS: We conducted a prospective, examiner-masked, parallel-group study and randomized 38 subjects (age range 6 months to 12 years, mean = 7.3 years) into four groups such that the same combination mydriatic-cycloplegic solution was administered under the following conditions: 1) eyedrops to eyes that were open; 2) eyedrops to eyes that were closed; 3) spray to eyes that were open; 4) spray to eyes that were closed. Both eyes of each subject received the solutions administered under one of the four conditions. Residual accommodation was measured using dynamic retinoscopy and the subjective push-up method, where applicable, at times 0, 10, 20, 30, 45, and 60 mins following administration of the medications. RESULTS: Repeated measures analysis of covariance revealed no statistically significant difference (p > 0.05) in cycloplegic efficacy among the four methods of drug delivery. CONCLUSIONS: The results of this study suggest that administering cycloplegics by spray to the closed eye is as efficacious as instilling eyedrops to the open eye. Use of a cycloplegic spray can have important clinical advantages compared to conventional cycloplegic eyedrops administered to children. PMID- 8227938 TI - Mydriatic efficacy of a cycloplegic spray in the pediatric population. AB - BACKGROUND: The topical administration of eyedrops is often an unpleasant procedure in the pediatric population. We sought to learn whether a combination mydriatic-cycloplegic solution would be an effective mydriatic if applied as a spray. METHODS: We used a prospective, examiner-masked, parallel-group study design and randomized 38 subjects (age range 6 months-12 years, mean = 7.3 years) into four groups such that the same combination mydriatic-cycloplegic solution was administered under the following conditions: 1) eyedrops to eyes that were closed; 2) eyedrops to eyes that were open; 3) spray to eyes that were closed; 4) spray to eyes that were open. Both eyes of each subject received the solutions administered under one of the four conditions. Photographs were taken of each subject's pupils at times 0, 5, 15, 25, 40, and 55 mins following administration of the medications. RESULTS: Repeated measures analysis of variance revealed no statistically significant difference (p > 0.05) in mydriatic efficacy among the four methods of drug administration. CONCLUSIONS: The results of this study suggest that the mydriatic efficacy of a mydriatic-cycloplegic spray is comparable to that associated with eyedrop instillation. Mydriatic sprays may have important clinical advantages over conventional ophthalmic solutions in the pediatric population. PMID- 8227939 TI - Types of optometric testing emphasized in different practice modes. AB - BACKGROUND: In order to determine if tests performed in an examination vary with practice settings, we surveyed all 5,200 California licensed optometrists and asked them to identify their primary mode of practice and the percentage of time and/or frequency that certain test procedures were done in an eye examination. METHODS: We restricted our analysis to three practice modes: private, corporate, and nonprofit/health maintenance organization (NP/HMO). We received a 66.4 percent response rate. To determine any trends in emphasis of testing by the three practice modes and to establish relationships between the three, we compared each mode of practice to the other. The combinations were private vs. corporate, corporate vs. NP/HMO, and private vs. NP/HMO. The three combinations were compared to the frequency of performing case history, stereo acuity testing, binocular vision testing, biomicroscopy, dilated fundus examination, tonometry and perimetry. RESULTS AND CONCLUSIONS: We found the following relationships in the frequency/emphasis of testing as statistically significant: 1) For stereopsis testing: private practices performed it more often than corporate; 2) For binocular vision testing: private practices performed it more than did either corporate or NP/HMO, corporate performed it more than NP/HMO; 3) For tonometry: corporate performed it slightly more than NP/HMO; 4) For dilation: NP/HMO performed it more than both private and corporate; and 5) For perimetry: private performed it more than both corporate and NP/HMO. PMID- 8227940 TI - A new NMR airlift bioreactor used in 31P-NMR studies of itaconic acid producing Aspergillus terreus. AB - An airlift bioreactor for in-vivo NMR studies of cells is described. The 10-mm diameter airlift reactor was constructed for studies of mycelial/pellet forming organisms, grown in suspension. With this device 161 MHz 31P-NMR spectra of living Aspergillus terreus cells, producing itaconic acid, have been obtained. Signals were observed for intra- and extracellular orthophosphate, glycerol-3 phosphorylethanolamine (GPE), glycerol-3-phosphorylcholine (GPC), sugar phosphates and polyphosphate. The spectra also showed broad overlapping resonances in the shift range of NAD(H) and NADP(H). Polyphosphate disappeared when the respiratory gas was exchanged for pure N2. The intracellular pH was estimated at 6.2. In spectra of cell extracts approx. 60 peaks were observed in the range of 20 to -22 ppm, and they confirmed the appearance of the metabolites observed in living cells. PMID- 8227941 TI - Determination of immune complexes by high-performance gel chromatography (positive cooperativity of antibody-antigen reaction). AB - Positive cooperativity of antibody-antigen complex formation using human Cu/Zn superoxide dismutase and a murine monoclonal antibody as model is demonstrated by high-performance gel chromatography using TSK G3000sw and TSK G4000sw columns. Three different antibody-antigen complexes named Type I, II and III composed from two antigen molecules+two antibody molecules, two antigen molecules+one antibody molecule and one antigen molecule+one antibody molecule could be separated. The degree and ratio of complexes present in solution depends mainly on the ratios of antibody and antigen. PMID- 8227942 TI - Reduced binding capacity of concanavalin A-sepharose after treatment with chaotropic agents. AB - Affinity chromatography on Concanavalin A-Sepharose is widely used for purification of glycoproteins carrying mannose and glucose oligosaccharide moieties. High concentrations of chaotropes elute Concanavalin A from the gel matrix and significantly reduce the binding capacity of the column. PMID- 8227943 TI - Lipophilicity determination of non-homologous series of nonionic surfactants by means of reversed-phase thin-layer chromatography. AB - The lipophilicity and specific hydrophobic surface area of 22 nonionic surfactants having different hydrophobic moieties have been determined by reversed-phase thin-layer chromatography using methanol as an organic modifier. RM values decreased linearly with increasing concentration of methanol. Both the lipophilicity and specific hydrophobic surface area of the surfactants decreased with increasing numbers of ethylene oxide groups per molecule. The significant correlation between the RM0 value and the specific hydrophobic surface area indicates that these surfactants can be considered as a homologous series of solutes independently of their structural inhomogeneity. Stepwise regression analysis suggested that the hydrophobic moieties are not entirely solvated and they are in a folded state in the eluents. PMID- 8227944 TI - Determination of singlet oxygen quantum yields with 1,3-diphenylisobenzofuran in model membrane systems. AB - The oxidation of 1,3-diphenylisobenzofuran by singlet oxygen was investigated in methanol and in two different types of liposomes. It was found that at high concentrations of scavenger 1,3-diphenylisobenzofuran, e.g., > 100 microM in methanol, the 1:1 oxidation stoichiometry is lost and more than one scavenger molecule per molecule of singlet oxygen is consumed. In model membrane systems, where local scavenger concentrations are high due to compartmentalization, correct singlet oxygen quantum yields with 1,3-diphenylisobenzofuran are only determined if the increased oxidation is taken into account. PMID- 8227945 TI - The substrate-depletion error on the multiple Michaelis-Menten equation with and without an added term linearly dependent on the substrate concentration. AB - Saturation curves for substrate interaction with an acceptor (enzyme, binding or carrier protein) are often analysed on the assumption that the amount of acceptor bound substrate is negligible compared to its total amount. Analytical criteria permitting one to decide whether the assumption is justified or not for systems described by a single Michaelis-Menten equation have been derived previously. For more complicated systems, error formulae often cannot be obtained in closed form, and, if obtainable, are unwieldy. How this more complicated problem can be tackled is shown for mixtures of acceptor sites described by a sum of several Michaelis-Menten terms, without or with an additional term for 'non-specific' uptake or binding. In particular, it is shown that the maximum error, for which simple analytical expressions are obtained, provides a valid criterion for assessing whether substrate depletion is negligible or not. PMID- 8227946 TI - Do the spectra of maleimide spin-labelled whole blood platelets reflect the structure and conformation of membrane proteins? AB - The maleimide spin label (4-maleimido-2,2,6,6-tetramethylpiperidine-1-oxyl, MSL), the commonly used reagent specific for cysteine thiol groups in proteins, penetrates cell membranes and binds to both the membrane and cytoplasmic protein moieties. In order to differentiate the labelling of these two subpopulations of cell proteins, we developed three different methodological approaches varying in the consequent distribution of this label in platelets. (A) The labelling of platelet proteins was negligible when bovine serum albumin was used in the Tyrode's buffer for the isolation of platelets, as the majority of the spin label was bound to the albumin coated on the platelets. (B) Preblocking of the reactive thiol groups in albumin with non-spin maleimide analog, N-ethylmaleimide (NEM), caused a considerable amount of MSL to bind with whole platelets but the impartment of membrane component was below 50%. It suggests that the majority of the spin label penetrated platelets and was bound to the intrinsic platelet proteins. (C) In order to prevent labelling of intrinsic platelet proteins with MSL, platelets were preincubated with N-ethylmaleimide, which was able to penetrate platelets and block the reactive thiol groups inside the cells. Such a treatment resulted in a saturation of the intrinsic protein residues with this non-spin analog. The subsequent incubation of thus-treated albumin-free platelets with MSL was to enhance considerably the likelihood of the attachment of MSL molecules to the thiol groups available in platelet-membrane proteins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227947 TI - Usefulness of difference spectroscopy in the study of the binding of uracil derivatives to human serum albumin. AB - Propylthiouracil is an antithyroid drug which is carried by the blood, thanks to its binding with human seroalbumin (HSA), and induces a structural alteration in HSA that changes the binding capability of other ligands. Then, with the aim of fixing the functional group in propylthiouracil involved in the interactions with HSA, the binding parameters for several uracil derivatives bound on HSA have been estimated. Interaction of propyluracil, thiouracil and propylthiouracil with HSA leads to the formation of complexes that show spectral shifts. These spectral shifts are a measure of the fraction of chromophoric groups, which is perturbed in the interaction with HSA, and thus can be used in the ligand binding estimate. The difference spectroscopy results correspond to the binding on a single centre in HSA. The difference spectra of propyluracils in seroalbumin coincide with those of propyluracils in a perturbant solvent (ethanol). On the other hand, propylthiouracil and propyluracil bind to seroalbumin on a larger scale than uracil and thiouracil. Thus, we can conclude that this binding is strengthened by hydrophobic interactions between the propyl group in propyluracils and apolar substituents on HSA. PMID- 8227948 TI - One-step immunoaffinity purification of human beta 1 thyroid hormone receptor with DNA and hormone binding activity. AB - An efficient and versatile method to purify large amounts of active human beta 1 thyroid hormone receptor (h-TR beta 1) was developed. Using a T7 expression system, h-TR beta 1 was overexpressed in Escherichia coli. Approx. 80% of the expressed receptor protein was concentrated in the insoluble inclusion bodies and approximately 20% was in the soluble form (h-TR beta 1-S). h-TR beta 1-S was conveniently purified by one immunoaffinity chromatographic step. From 1 l of cell culture, approx. 0.1 mg of purified h-TR beta 1-S was obtained. The purified h-TR beta 1-S binds to 3,3',5-triiodo-L-thyronine with a Ka = 2 x 10(9) M-1 and exhibits analog specificity. The purified h-TR beta 1-S also binds to T3 response elements (TRE) with different orientation in the half-sites with differential activity. In addition, binding of h-TR beta 1-S to TREs was enhanced by retinoid X receptor. These results indicate that the purified h-TR beta 1-S retains its hormone and DNA binding activity. The purified h-TR beta 1-S is suitable for structural and functional studies. This method could be used to purify h-TR beta 1 or rat TR beta 1 expressed in insect cells or yeast. PMID- 8227949 TI - Power spectral analysis of heart rate and arterial blood pressure oscillation in brain-dead patients. AB - To clarify the mechanism of the slow arterial blood pressure oscillation seen in brain-dead patients, we investigated the frequency of fluctuations in arterial blood pressure and heart rate using power spectral analysis. The electrocardiogram, arterial blood pressure and respiration were recorded simultaneously from 9 brain-dead patients and 8 vegetative patients. Power spectral analysis of these data revealed a very slow fluctuation (0.002-0.01 Hz) in arterial blood pressure in brain-dead patients, the frequency of which was equal to that of the low-frequency spectrum of heart rate, indicating vasomotor sympathetic activity. Neuropathological examinations of the medulla and spinal cords of 4 autopsied brain-dead patients revealed that the spinal cord, ventral and dorsal nerve roots, and the nucleus intermediolateralis of the lateral horn below the level of C3/4 were virtually intact. These findings suggest strongly that the slow oscillation of arterial blood pressure in brain-dead patients originates from the vasomotor tone controlled by spinal sympathetic nerves. PMID- 8227950 TI - Neuronal metabolism of catecholamines: plasma DHPG, DOMA and DOPAC. AB - Pre-synaptic endings of the sympathetic nervous fibers control the metabolism of catecholamines, particularly inactivating norepinephrine after its neuronal recapture. The present study was carried out to investigate this segment of the metabolism of catecholamines through measurements of DHPG, DOMA and DOPAC concentrations in plasma. A sensitive and specific radio-enzymatic assay was developed of which the major characteristic is to include the plasma sample in the incubation mixture without initial extraction of the deaminated metabolites. In the rat, there was a statistically significant correlation between norepinephrine and DHPG in both anesthetized and conscious conditions and after clonidine or guanethidine induced reduction of sympathetic activity; thus it can be suggested that plasma DHPG is a good index of neuronal metabolism of norepinephrine in this animal. In humans, our data indicate an interesting correlation between norepinephrine and DOMA concentrations in plasma in resting conditions and within three hours after clonidine. Further studies need to be carried out to establish whether DOMA is a better index of neuronal metabolism of norepinephrine than DHPG. PMID- 8227951 TI - Topographical distribution and immunocytochemical features of colonic neurons that project to the cranial mesenteric ganglion in the pig. AB - Using the retrograde neuronal tracers Fast blue and Fluorogold, the topographical distribution and morphological features of porcine colonic neurons projecting to the cranial (superior) mesenteric ganglion have been investigated. Two to four weeks after injection of the tracer into the cranial mesenteric ganglion of immature pigs, labelled neurons were found throughout the colon. In the myenteric and outer submucous plexuses, they were present in ganglia situated to the side of the mesenteric attachment. The highest density of labelled neurons was observed at the end of the ascending colon, which in the pig represents 78-80% of the total colon length. The viscerofugal neurons had a multidendritic appearance and part of them were immunoreactive for calcitonin gene-related peptide or serotonin. This study has revealed similarities but also significant differences in the colono-sympathico-colonic pathways between the pig and small laboratory animals such as the guinea-pig. PMID- 8227952 TI - Intramucosal nerve cells in human small intestine. AB - We investigated the intramucosal nerve cells of the human small intestine with histochemical methods to demonstrate nicotinamide adenine dinucleotide diaphorase and acetylcholinesterase and with morphometry. Intramucosal neurons appeared as solitary cells or in small groups, especially in the ileum. Most intramucosal nerve cell bodies were round or oval; some were flat or spindle-shaped. They mostly lay close to the muscularis mucosae, but some were located within the muscularis mucosae and others were some distance away from it. The processes of some mucosal neurons projected towards the submucosa. Most mucosal nerve cells showed acetylcholinesterase activity. The frequency distribution of nerve cell profile areas in the intramucosal cells in the duodenum differed from that of cells in the ileum (P < 0.001). There were more large mucosal nerve cells in the mucosa of the duodenum than in the ileum. There was no significant difference between the frequency distributions of cell profile areas of cells of the mucosa and cells of Meissner's and Henle's plexuses in the same region. We conclude that intramucosal nerve cells, similar to those of the submucosal plexus, exist in the human small intestine. The size of intramucosal nerve cell profiles differs between the duodenum and ileum. This is consistent with their possible different functions. PMID- 8227953 TI - Discharge pattern of neurons in the nucleus tractus solitarii (NTS): its cardiac rhythm is modulated by firing rate of the neurons. AB - In the nucleus tractus solitarii (NTS) neurons discharge in relation to cardiac rhythm. This cardiac rhythm exhibits various patterns designated as CRDPs (cardiac rhythmic discharge patterns). The CRDPs are estimated by post-event-time histograms (PETH) triggered by the R-waves of the ECG. Modulations of CRDPs appear as changes in the number and height of peaks in the PETHs. The amount of basic activity, which is not related to the cardiac cycle, alters CRDP. PETHs constructed during various phases of respiration reveal modulations of CRDPs within the respiratory cycle. As our previous work indicated, the NTS neurons exhibit typical reticular rhythms. In this paper we also found that the basic activity of NTS neurons was often changed by other influences for which no comparable patterns could be observed in other simultaneously acquired signals. When we constructed PETHs according to the activity level of the NTS neurons, i.e., firing level per cardiac cycle, modulations of CRDPs which were even stronger than respiratory or reticular rhythmical modulations became clear. The modulations of CRDPs caused by different origins were found to be present in the same neuron interlaced in time. The possible role played by these modulations of CRDPs in the coordination of different functional systems in the organism is discussed. PMID- 8227954 TI - Neuropeptide Y potentiates noradrenaline-evoked vasoconstriction by an intracellular calcium-dependent mechanism. AB - The potentiating effect of neuropeptide Y (NPY) was examined by testing the influence of putative inhibitors of calcium entry on the NPY-enhanced contractile response to noradrenaline in the guinea pig uterine artery. In order to examine the involvement of voltage sensitive calcium entry mechanisms we recorded the effect of noradrenaline and NPY on the membrane potential. NPY (100-300 nM) enhanced noradrenaline-evoked vasoconstriction. The potentiation by NPY was most prominent in low noradrenaline concentrations (30-300 nM) and the pD10 (-log molar concentration of agonist eliciting 10% of maximum contraction) value was increased from 6.43 +/- 0.07 to 6.97 +/- 0.11 (P < 0.001, n = 6). Inhibition of extracellular calcium influx shifted concentration-dependently to the right the concentration-response curve for noradrenaline but potentiation by NPY still remained. The intracellular calcium chelator quin-2 AM selectively abolished the NPY-induced enhancement of the contractile response to noradrenaline. In contrast, quin-2 AM (10-30 microM) had no inhibitory effect on the contractile response to noradrenaline per se. It is suggested that NPY initiates an intracellular calcium-sensitive mechanism which increase alpha-adrenoceptor sensitivity. This results in a significant increase of sarcoplasmic calcium and stronger contractile responses to noradrenaline. PMID- 8227955 TI - Long-term hypoxia increases the number of norepinephrine-containing glomus cells in the rat carotid body: a correlative immunocytochemical and biochemical study. AB - A group of adult rats was divided into two subgroups: one was submitted to long term normobaric hypoxia (10% O2, two weeks) while the other (control group) was kept in the same room, breathing air. Animals from each subgroup were used to study either the norepinephrine content of the carotid body by high pressure liquid chromatography or to localize norepinephrine-containing structures using an immunocytochemical procedure (peroxidase-labelled antibodies on cryostat sections). The biochemical study showed, as expected, a large increase in carotid body norepinephrine content (19-fold) and turnover (ten-fold) in hypoxic rats. The immunocytochemical study revealed only a few norepinephrine-immunopositive glomus cells in sections through the carotid body of normoxic rats, whereas the carotid body of hypoxic rats showed a very large number of norepinephrine positive glomus cells. This increase was quantified, using an image analyser, and it was found to constitute a 61-fold increase in the number of immunopositive profiles per section and a 29-fold increase in the immunopositive profile area/section area ratio. It is concluded that long-term hypoxia increases rat carotid body norepinephrine content by inducing norepinephrine synthesis in glomus cells in which this amine was not detectable previously, before hypoxia. PMID- 8227956 TI - A role for serotonin in the elaboration of a differential pattern of activity in sympathetic nerves to kidney and skeletal muscle vasculature. AB - The role of serotonin (5-HT) in the elaboration of patterns of regional sympathetic discharge evoked from the ventral medulla was examined in cats under Saffan anesthesia. A differential response pattern consisting of an increase in renal sympathetic nerve activity simultaneous with no change in activity to skeletal muscle vasculature was evoked from a group of ventral medullary neurons by microinjection of D,L-homocysteic acid. Subsequent intrathecal administration of the 5-HT receptor antagonist methysergide (200 micrograms) attenuated or abolished the excitatory response to the kidney on restimulation of the medulla, without affecting sympathetic activity to muscle. Methysergide did not affect an excitatory response evoked in the outflow to both the kidney and skeletal muscle vasculature from an adjacent region of the ventral medulla. These results indicate a role for 5-HT in the elaboration of a specific differential pattern of sympathetic response evoked from medullary neurons. PMID- 8227957 TI - The localization of sympathetic and vagal neurones innervating the carotid sinus in the rabbit. AB - The distribution of neurones innervating the carotid sinus of the rabbit was determined by inserting chips of horseradish peroxidase (HRP) into the adventitial layer of the carotid sinus. Labelled neurones were found in the superior cervical ganglion, along the cervical sympathetic trunk and in the nodose ganglion (mean counts of 1876, 139 and 232, respectively). No labelled neurones were located in the middle cervical ganglion or the stellate ganglion. Within the superior cervical ganglion, the fraction of labelled neurones found in equal longitudinal quadrants of the ganglion were, from rostral to caudal, 2%, 12%, 42% and 44%. Thus the majority of neurones were located in the caudal region of the superior cervical ganglion. The primary pathway for sympathetic fibres innervating the carotid sinus was shown to be in branches of the external carotid nerve. The greatest number of labelled neurones in the nodose ganglion were located in the most rostral quadrants of the ganglion. PMID- 8227958 TI - Calcitonin gene-related peptide innervation and binding sites in rat aorta during development. AB - Indirect immunohistochemistry performed on whole mounts of arch and thoracic part of the rat aorta at six developmental stages (from embryonic day 17 to 6 months, in males and females) revealed that calcitonin gene-related peptide (CGRP) innervation is highest in the arch. The highest density of innervation is found at the three first postnatal ages investigated (day 1, day 3 and 5 weeks; 2.6 +/- 0.6 intercepts/mm in the arch at 1 day); however, all values are low compared to other arteries. The innervation grows from a few short isolated fibres in the embryo to a more complex meshwork in older animals. No striking differences were noticed between males and females. Autoradiographic studies were performed on serial sections at several levels of the aorta but did not reveal binding sites for CGRP in the vascular wall. This might be due to the technique which does not allow visualization of low density of binding sites, or to binding sites of weak affinity. We discuss the possible importance of CGRP in rat aortic smooth muscle development. PMID- 8227959 TI - Autonomic and hemodynamic responses and interactions during the Mueller maneuver in humans. AB - We compared the responses to a Mueller maneuver maintained for 20 s to effects of an equal period of end expiratory apnea. We measured heart rate, mean blood pressure (BP), central venous pressure (CVP), and sympathetic nerve activity (SNA) in 9 normal humans. The Mueller maneuver was accompanied by a fall in CVP from 5 +/- 1.2 to -13 +/- 3.2 mmHg (P < 0.05). During the first 10 s of Mueller, BP fell from 95 +/- 4.2 to 81 +/- 5.5 mmHg and SNA fell as low as 16 +/- 6% of control (P < 0.05). For the 5 s prior to release SNA increased to 236 +/- 36% (P < 0.05), and BP began to increase. Release of the Mueller resulted in a surge in BP to 104 +/- 5.8 mmHg and suppression of SNA to 61 +/- 48% (P < 0.05). By contrast, there was no fall in BP or CVP during apnea and SNA increased to 188 +/ 24% for the first 5 s. Between 16 and 20 s of apnea SNA was 231 +/- 52% and BP increased from 92 +/- 3.1 to 96 +/- 3.6 mmHg (P < 0.05). Release of apnea resulted in a surge in BP to 105 +/- 3.0 mmHg and suppression of SNA to 30 +/- 12% (P < 0.05). Oscillations in BP and SNA during the Mueller maneuver may contribute to similar oscillations, and hence cardiovascular consequences, in patients with sleep apnea. PMID- 8227960 TI - Naloxone does not prevent vasovagal syncope during simulated orthostasis in humans. AB - The mechanism of vasovagal syncope during orthostasis in humans is unknown. Opioid receptors have been implicated in the vasovagal-like responses to hemorrhagic hypotension in conscious animals. We sought to determine if opioid receptor blockade with naloxone (mu receptor antagonist) would prevent or delay the onset of vasovagal syncope in humans. Three protocols were performed in which heart rate, arterial pressure, sympathetic nerve activity, thoracic impedance and forearm vascular resistance were measured during stepwise steady-state increments of lower body negative pressure (LBNP) in nine healthy volunteers. In protocol 1, duplicate trials of LBNP to syncope or -60 mmHg were performed with a 30-45 minute rest period separating the trials. No significant differences in any physiologic responses or cumulative stress tolerance were found. In protocol 2, graded LBNP was repeated after administration of saline or naloxone (0.1 mg/kg) in six subjects in which vasovagal syncope occurred prior to -60 mmHg LBNP. The peak increase of sympathetic nerve activity during LBNP was augmented after naloxone (P = 0.02), but the occurrence of vasovagal syncope was not prevented nor was the cumulative stress tolerated affected (P = 0.42). The heart rate and arterial pressure responses to LBNP were not affected by naloxone. Similarly, in protocol 3, naloxone given just prior to the onset of pre-syncopal symptoms did not alter the physiologic response or the occurrence of vasovagal syncope. These data show that naloxone does not prevent or delay the onset of vasovagal syncope in humans which suggests that mu opioid receptors do not mediate the vasovagal response. PMID- 8227961 TI - Effects of vasopressin on the response to carotid occlusion in conscious rabbits. AB - To investigate the interaction between arginine vasopressin and the carotid sinus baroreflex, hemodynamic responses to bilateral carotid occlusion and to controlled reductions in carotid sinus pressure were examined. In the control state and during vasopressin infusion in conscious rabbits, mean arterial pressure, heart rate, mean aortic flow and total peripheral resistance were measured. Vasopressin infusion at 5 or 10 ng/kg/min did not raise arterial pressure, but increased resistance, and decreased heart rate and aortic flow in a dose-dependent manner. The pressure and resistance responses to carotid occlusion or changes in carotid pressure were not altered by vasopressin. The heart rate response was augmented significantly from 23 +/- 5 (mean +/- S.E.) to 40 +/- 8 and 43 +/- 8 beats/min with infusion of 5 and 10 ng/kg/min of vasopressin. Vasopressin did not augment the gain of carotid sinus reflex control of arterial pressure (3.7 +/- 0.5 in control and 3.5 +/- 0.5 during 5 ng/kg/min of vasopressin). With vasopressin infusion at 5 ng/kg/min, following vagal blockade with methylatropine both the arterial pressure and the heart rate responses to carotid pressure changes decreased to 73% and 32% of the response before blockade. The data indicate that vasopressin has little effect on control of arterial pressure by the carotid sinus baroreflex in conscious rabbits when vagal responses are activated. PMID- 8227962 TI - Desensitization by neuropeptide Y of effects of sympathetic stimulation on cardiac vagal action in anaesthetised dogs. AB - The effects of a long-lasting intravenous infusion of neuropeptide Y (NPY, 180 +/ 8 min, 53 +/- 4 micrograms/kg/h) on the prolonged inhibition of cardiac vagal action evoked by cardiac sympathetic nerve stimulation and bolus intravenous injections of NPY were investigated in anaesthetised dogs. Sympathetic stimulation and NPY injection were performed on four separate occasions; once in control conditions, then once early and again late in the period of NPY infusion, and then on a final occasion 60-90 min after the cessation of NPY infusion. The maximum inhibition of cardiac vagal action evoked by an injection of NPY was significantly less late in the NPY infusion when compared with the other three injection groups (ANOVA, P < 0.001). Also the time to half-recovery of this response was significantly less than that seen in the other three injection groups (ANOVA, P < 0.001). The maximum inhibition of cardiac vagal action evoked by sympathetic stimulation was significantly reduced late in the NPY infusion when compared with the other three stimulation groups (ANOVA, P < 0.0001). The time for half-recovery of this response was also less than that of the other three stimulation groups (ANOVA, P < 0.001). The results indicate that desensitisation of the vagal attenuation to both exogenous NPY and sympathetic stimulation occurred during a long-lasting period of NPY infusion. This is consistent with the proposal that NPY is a mediator of this sympathetic-evoked vagal attenuation. PMID- 8227963 TI - Effects of 5-hydroxytryptamine on discharge of vagal mucosal afferent fibres from the upper gastrointestinal tract of the ferret. AB - This study was undertaken to determine the effects of 5-hydroxytryptamine (5-HT) on vagal mucosal afferent endings and how this may relate to their sensitivity to other stimuli. Single afferent fibres with receptive fields in the mucosa of the upper gastrointestinal tract were recorded from the cervical vagus of Urethane anaesthetized ferrets. The selection criteria included failure to respond to luminal distension (i.e., vagal tension receptors were excluded). All fibres tested responded to mucosal stroking. The majority of these (28/32) also responded to close-intrarterially applied 5-HT (10 micrograms) with a brief burst (usually < 15 s) of action potentials, which in 6/6 cases was reduced or abolished by the 5-HT3 receptor antagonist granisetron (0.1-0.5 mg/kg i.v.), as were four responses to cholecystokinin-octapeptide (100-400 pmol close I.A.). The response to 5-HT was shown to be dose-dependent over the dose range 2-75 micrograms on six occasions. Responses to luminal stimuli, which included 150 mM HCl, 1 M NaCl, and mucosal stroking, were not blocked by granisetron although in three fibres, the resting discharge was reduced by the antagonist, suggesting that resting discharge in vagal mucosal afferents may be influenced by endogenously released 5-HT. PMID- 8227964 TI - Effects of 5-hydroxytryptamine (5-HT) on the discharge of vagal mechanoreceptors and motility in the upper gastrointestinal tract of the ferret. AB - This study was undertaken to investigate the sensitivity of vagal mechanoreceptor afferent fibres from the stomach and small intestine to 5-hydroxytryptamine (5 HT, 10 and 50 micrograms), and to relate this to motor activity. The discharge of single afferent fibres was recorded from the cervical vagus of Urethane anaesthetized ferrets. These had receptive fields in the corpus (15 fibres), antrum (11 fibres), duodenum (3 fibres) or jejunum (1 fibre), and their discharge was excited by distension of the segment containing the receptive field, or by mechanical probing. Conduction velocity was calculated in 7 fibres, and always fell in the C-fibre range (0.84 +/- 0.07 m/s). Injection of 5-HT intra-arterially via the coeliac axis evoked both contraction and relaxation with the dominant effect dependent on region and dose of 5-HT. However, irrespective of the motor response, the mechanoreceptor discharge closely mirrored both the magnitude and the time-course of the motor response. The relationship between the changes in discharge of individual mechanoreceptors and change in intraluminal pressure in all regions in response to both doses of 5-HT fitted a linear regression y = 17.1 +/- 1.2x and was highly significant (P < 0.0001), indicating that 5-HT has no modulatory effect on the sensitivity of mechanoreceptors to changes in intraluminal pressure. This study suggests that vagal mechanoreceptors supplying the gastrointestinal tract respond to changes in motor activity evoked by 5-HT, rather than to 5-HT directly. PMID- 8227965 TI - Contribution of bed nucleus of the stria terminalis to the cardiovascular responses elicited by stimulation of the amygdala. AB - Anatomical and physiological studies were done in the rat to investigate the possibility that the cardiovascular responses elicited by stimulation of central nucleus of the amygdala (ACe) were mediated via projections to bed nucleus of the stria terminalis (BST). In the first series, to determine the distribution of neurons in ACe that projected to the cardiovascular region of BST, the retrograde tracer Fluorogold (FG) or rhodamine latex micro-beads (Rd) were injected into BST. FG and Rd injections that overlapped the cardiovascular region of BST resulted in retrogradely labelled neurons throughout the amygdala. In ACe, retrogradely labelled neurons were observed primarily in the lateral subdivision of the rostral ACe compared to the caudal ACe. The medial subdivision of ACe was found to have very few retrogradely labelled neurons. In the second series, the effect of either blocking synaptic transmission in BST with CoCl2, chemical lesions of BST with ibotenic acid (IBO), or electrolytic lesions of BST on the depressor response elicited by either electrical or chemical stimulation of ACe was investigated in the chloralose-anesthetized, artificially ventilated and paralysed rat. Microinjections of CoCl2 into BST significantly attenuated the depressor responses to stimulation of the rostral components of the lateral subnucleus of ACe, but not those to stimulation of the caudal and medial components of ACe. Microinjections of IBO into BST or electrolytic lesions of BST resulted in similar effects on the depressor responses to ACe stimulation. Taken together, these data indicate that neurons within the rostral components of the lateral subnucleus of ACe project to the cardiovascular region of BST and mediate in part the depressor responses to stimulation of the rostral ACe. On the other hand, the depressor responses elicited from the caudal ACe are not mediated through BST. These results suggest that at least two independent pathways originate in the ACe that influence the circulation. PMID- 8227966 TI - Role of the central and arterial chemoreceptors in the response of gastric tone and motility to hypoxia, hypercapnia and hypocapnia in rats. AB - The contribution of autonomic nerve activity to stomach tone and motility during central and arterial chemoreceptor excitation or inhibition was analyzed in urethane anesthetized, artificially ventilated rats. Systemic severe hypoxia at end-tidal O2 concentration (FETO2) 6% and systemic hypercapnia at end-tidal CO2 concentration (FETCO2) 6%, 8% and 10% applied for 1 min produced a significant depression in gastric tone and motility. Hypocapnia at 3% FETCO2 increased gastric tone and motility. Hypoxia co-activated both the sympathetic and the vagal efferent gastric nerve branches. Hypercapnia augmented only sympathetic gastric efferent nerve activity but not vagal efferent nerve activity. Hypocapnia slightly increased vagal nerve activity to the stomach. Bilateral denervation of the arterial chemoreceptors significantly attenuated the inhibitory gastric response to hypoxia. Similar attenuation of hypoxia-induced depression of gastric tone and motility was produced by bilateral gastric sympathectomy but not by vagotomy. In contrast, the inhibitory effect of severe hypercapnia and the facilitatory effect of hypocapnia upon gastric tone and motility were unaffected by arterial chemoreceptor denervation, by severance of gastric sympathetic branches or by gastric vagal denervation. Hyperoxia at 90% FETO2 had no effect on the gastric nerve activities, gastric tone or motility. It is concluded that in the rat hypoxia co-activates sympathetic and vagal efferent nerve activities to the stomach via an arterial chemoreceptor reflex, and that hypercapnia activates sympathetic gastric nerve activity via central chemoreceptors. Hypocapnia activates efferent vagal gastric nerve activity. All chemical stimuli except that of hyperoxia have a significant local effect on the gastric tone and motility.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227967 TI - Delayed-type skin allergic reaction in guinea pigs induced by anti-rheumatic compounds with sulfhydryl groups. AB - Delayed-type skin allergic (DTA) reactions induced by various sulfhydryl compounds were investigated. Sulfhydryl compounds investigated were bucillamine and D-penicillamine: antirheumatic agents, SA981: an intramolecular cyclic disulfide of bucillamine, tiopronin: a hepatoprotective and potent antirheumatic agent and captopril: antihypertensive and potent antirheumatic agent. Guinea pigs were sensitized on day 1 by subcutaneous injection and day 7 by subcutaneous and intramuscular injection of emulsions of these compounds (3 mg/animal) with Freund's complete adjuvant. Two weeks after the second sensitization, 0.3 mg/animal of each compound was intradermally challenged and the 24 hr DTA reaction was evaluated. A large number of sulfhydril compounds showed positive in the DTA reactions and the intensity of these reactions had a good correlation with the frequency of the skin rashes referred to by the adverse effects in clinical reports of anti-rheumatic drugs. It is suggested that guinea pig DTA reaction may provide a good screening way to find new sulfhydryl or disulfide compounds with low incidence of skin rash. A novel sulfhydryl compound N-(2,2 dimethyl-3-mercaptopropionyl)-L-cysteine (SA3441) and its intramolecular cyclic disulfide (4R)-hexahydro-7,7-dimethyl-6-oxo-1,2,5-dithiazocine-4-carboxylic acid (SA3443), which show no DTA activity in guinea pigs, were found using the above mentioned DTA reaction. PMID- 8227968 TI - Iron polymers impair the function and maturation of macrophages. AB - An inhibitory effect of iron salts on various immune functions in vitro has been reported in several laboratories during the last years. This work extends such observations by showing that iron citrate, but not sodium citrate, inhibits the function and maturation of murine macrophages (MOs). However, such inhibition is only observed in the presence of ferric citrate with a metal-to-ligand molar ratio of 1:1, but not with ferric citrate with a metal-to-ligand molar ratio of 1:10 in which the hydrolyzation and polymerization of iron in physiological solutions is prevented. Accumulation of ferric iron on the cytoplasm of MOs was observed, but only in the group of MOs treated with ferric citrate 1:1. Increasing the concentration of serum in the culture medium diminished the inhibitory effect of ferric citrate 1:1. The inhibitory capacity of iron polymer was probably associated to its ability to both interact with the cell constituents of the cytoplasm and stimulate lipid peroxidation. PMID- 8227969 TI - In vivo effects of alprazolam and lorazepam on the immune response in patients with migraine without aura. AB - Over the past few years, the immunomodulating role of benzodiazepines (BDZ) has been reported in literature. In particular, diazepam is an inhibitory BDZ with regard to its effects on the phagocytic and metabolic activities of polymorphonuclear cells (PMN) and monocytes, while triazolobenzodiazepines (alprazolam and triazolam) upregulate normal human peripheral blood T lymphocyte function. On these grounds, the administration of alprazolam (1 mg/per day for 1 month) in 13 patients with migraine without aura (MWA) and of lorazepam (2 mg/per day for 1 month) in 10 matched MWA subjects has been evaluated in terms of immune response. Results show that before administration of BDZ in both groups of patients phagocytosis and killing of PMN and monocytes were profoundly depressed and the same was true for the lymphocyte-dependent antibacterial activity. After one month treatment lorazepam further decreased lymphocyte function without modifying phagocytic capabilities. On the contrary, alprazolam increased PMN phagocytosis and killing and monocyte phagocytosis without modifying antibacterial activity values. Taken together, these results further support the existence of different classes of BDZ in terms of their immunomodulating capacities. Moreover, alprazolam seems to be a more appropriate BDZ for treating immunocompromised patients, even including MWA patients. PMID- 8227970 TI - VIP modulates intracellular calcium oscillations in human lymphoblasts. AB - Vasoactive intestinal polypeptide (VIP) has been shown to stimulate adenylate cyclase in a human lymphoblast cell line (MOLT 4). In the present study, we monitored fluorescence in cell suspensions and in single fura-2 loaded MOLT 4 lymphoblasts to determine if VIP modulates intracellular calcium concentrations ([Ca2+]i), and if this modulation is mediated by adenylate cyclase. The distribution of [Ca2+]i in resting and stimulated cells was non-homogeneous, with gradients of high [Ca2+]i present in the subplasmalemmal space. In a subset of cells (10-30% of all cells studied), [Ca2+]i showed La(3+)-sensitive, temporal changes in the form of [Ca2+]i oscillations with a baseline [Ca2+]i value of 115 +/- 10 nM, an oscillation amplitude of 150 +/- 18 nM and a mean period of 9.2 +/- 2 s. The remaining non-oscillating cells showed a constant [Ca2+]i level of 75 +/ 5 nM (n = 65 cells from 4 experiments). In the subset of cells with spontaneous [Ca2+]i oscillations, VIP dose-dependently (10(-12) to 10(-8) M) increased the amplitude of oscillations but did not stimulate their frequency. The stimulatory effect of VIP was correlated with baseline [Ca2+]i in these cells, was attenuated in the presence of La3+ (25 microM), but was unaffected by cell depolarization (126 mM KCl). Dibutyryl cyclic AMP (10(-4) to 10(-3) M) and forskolin (10(-4) M) had no effect on [Ca2+]i oscillations, or on [Ca2+]i in cells without oscillations. In cell suspensions, baseline [Ca2+]i was found to be 55.1 +/- 11.2 nM (mean +/- S.E.M., n = 11); VIP, cyclic AMP analogues or forskolin had no significant effect on [Ca2+]i. These findings suggest that: a) VIP modulates the amplitude of [Ca2+]i oscillations generated by a cytosolic [Ca2+] oscillator in a subset of cells at a concentration of 10(-12) M, a thousand-fold below the KD for the VIP receptor; b) baseline [Ca2+] values may be related to both the ability of cells to generate spontaneous [Ca2+] oscillations and of oscillating cells to respond to VIP; c) due to the small number of responding cells, VIP-induced [Ca2+]i changes are not detectable when studied in cell suspensions. PMID- 8227971 TI - Effectiveness of thymostimulin and study of lymphocyte-dependent antibacterial activity in children with recurrent respiratory infections. AB - Recurrent respiratory infections (RRI) consist of more relapsing acute respiratory infections than the ones expected for the age [> 6 acute respiratory tract infections (RTI) per year if age is > 3 years, and > 8 acute RTI per year if age is < 3 years]. Concerning the pathogenesis of RRI, several investigations report the important role of environmental factors, early socialization and immunological dysfunctions, such as lymphocyte subpopulations alterations, IgG subclass deficiency and phagocytosis and/or opsonization deficit during acute infections. In this framework, we have studied the lymphocyte-dependent antibacterial activity (ABA) among 121 children affected by RRI. Results show a statistically significant alteration of this function in 38 children (31.4%): 19 of them exhibited an absent ABA (group 1), while in the others same function was reduced (group 2). A bovine thymic extract, thymostimulin, was administered to both groups by intramuscular injections (1 mg/kg) for a 3 month cycle. At the end of therapy we observed a statistical significant rise of ABA only in group 1 and among children aged > 3 years. Among the same patients, 33 children (86.8%) improved in terms of reduction of clinical score and better results were seen among children aged > 3 years. These data emphasize the beneficial role of thymostimulin in RRI-affected children, suggesting a transient immaturity of the immune system as one of the possible pathogenetic factor. PMID- 8227972 TI - Antagonistic effects of endogenous and exogenous TGF-beta and TNF on auto-immune diseases in mice. AB - Injection of transforming growth factor beta 1 (TGF-beta 1) for five days during the late phase of the immunization process leading either to collagen type II induced arthritis (CIA) or to experimental allergic encephalomyelitis (EAE) protects against the development of these auto-immune diseases. Tumor necrosis factor alpha (TNF-alpha) injected during this same interval aggrevates CIA. In addition, anti-TGF-beta exacerbates and anti-TNF protects against CIA, acute and relapsing EAE, suggesting an important regulatory role for the endogenous production of the two cytokines on the severity of these diseases. More detailed studies about the mechanism of action of TGF-beta in acute EAE show that there is no detectable effect of TGF-beta on the development of sensitized T cells in vivo, as assayed by the proliferative responses of T cells from lymph nodes and peripheral blood to myelin antigens. Nevertheless, the number of lymphoid cells infiltrating the central nervous tissue is much greater in untreated than in TGF beta-treated, protected mice. We conclude that it is likely that TGF-beta protects against experimental auto-immune diseases by interfering with the entry of lymphoid cells into the target organs through inhibition of the upregulation of adhesion molecule expression on endothelial cells, and with subsequent inflammatory processes inside the target organs by antagonizing both the production and the effects of TNF. PMID- 8227973 TI - Cyclosporine in idiopathic nephrotic syndrome. AB - Idiopathic nephrotic syndrome encompasses two main forms of glomerular diseases, minimal change nephropathy and focal segmental glomerulosclerosis. Minimal change nephropathy is a disease of children which generally responds to corticosteroids. After remission, however, many patients show frequent relapses or steroid dependency. In these patients, cyclosporine may obtain remission of proteinuria in 80% of cases, although relapse usually occurs when the drug is stopped. Focal glomerulosclerosis is generally resistant to corticosteroids. Under cyclosporine some 40% of patients may attain complete or partial remission of the nephrotic syndrome particularly if low-dose prednisone is associated. Relapse of proteinuria usually occurs after stopping the drug. As cyclosporine may expose to chronic nephrotoxicity some guidelines should be followed to prevent this complication: - the doses should not exceed 5 mg/Kg/day - they should be adjusted whenever an increase in plasma creatinine of > or = 30% over the baseline values occurs - treatment should be stopped if there is no response within 3 months - a careful monitoring of patient under the supervision of a clinician trained with the use of cyclosporine is necessary. The term idiopathic nephrotic syndrome (INS) defines the association of a nephrotic syndrome with non specific glomerular lesions, in the absence of immune complex deposition (1). On the basis of renal histology two main types of INS are recognized: minimal change nephropathy (MCN) and focal and segmental glomerular sclerosis (FSGS). PMID- 8227974 TI - Immunomodulation due to coexposure to styrene and dioctyl phthalate in mice. AB - Pathomorphological and immunological alterations caused by a mixture of styrene and dioctyl phthalate were studied in albino mice following oral administration of 0.02, 0.03, 0.05 x LD50 of the mixture. The chemicals were mixed together proportionate to their respective LD50 values and fed in ground nut oil, 5 d/wk for 4 weeks. Histological examination of spleen revealed considerable depletion of cellular population of lymphoid follicles which corresponded to the dose dependent decrease in splenic mononuclear cell population count. The thymic lobules revealed slight atrophy but accompanied by a significant increase in thymocyte population. Correspondingly few significant histological changes were observed in mesenteric and peripheral lymph nodes. The treatment caused impairment of primary humoral immune response to SRBC (IgM) but there was a significant increase in response of splenocytes to B-cell mitogen LPS. There was a suppression of cutaneous delayed type hypersensitivity and increase in splenic lymphocyte response to T-cell mitogen PHA. Simultaneously, indirect immunity represented by decreased phagocytosis and enhanced metabolic function of reducing NBT by peritoneal exudate cells was observed. The in vitro exposure of vero cells to the mixture caused dose dependent protective effect. The results of present study indicate that subchronic exposure to low doses of mixture of styrene and dioctyl phthalate under certain conditions may modulate some of the immune functions as compared to exposure to either chemicals alone. PMID- 8227975 TI - Poly (adenosine diphosphate-ribose) synthesis in the anterior pituitary of the female rat throughout the estrous cycle: study of possible relation to cell proliferation and prolactin gene expression. AB - To investigate whether cell proliferation and PRL gene expression in female rat pituitary during estrous cycle were mediated by the poly (ADP)-ribosylation of chromatin proteins, anterior pituitaries at different estrous cycle were obtained from female Wistar rats, and poly (ADP-ribose) synthesis, DNA synthesis, PRL and GH messenger RNAs and PRL content in the pituitary, and serum concentrations of PRL and estradiol were analyzed. From diestrus to proestrus, poly (ADP-ribose) synthesis, the contents of PRL messenger RNA and PRL in the pituitary increased significantly, and decreased at estrus. However, DNA synthesis and serum concentration of PRL showed a significant increase from proestrus to estrus. Serum estradiol concentration increased from diestrus to proestrus. No significant change was observed in the pituitary GH messenger RNA content during estrous cycle. The increase of PRL messenger RNA from diestrus to proestrus was abolished completely by the administration of nicotinamide, an inhibitor of poly (ADP-ribose) synthesis, to rats at diestrus. These results indicate that poly (ADP)-ribosylation of chromatin proteins may play some role in cell proliferation and transcription of PRL gene during rat estrous cycle. PMID- 8227976 TI - Selenium administration does not cause thyroid insufficiency in subjects with mild iodine deficiency and sufficient selenium intake. AB - Selenium is a trace element essential for the activity of type I 5'-deiodinase which converts thyroxine (T4) to 3,5,3'-triiodothyronine (T3). In iodine deficient hypothyroid children at low selenium dietary intake the supplementation of selenium induced a significant decrement of serum FT4 and T4 concentrations and an increase of serum TSH concentrations. Since in western countries selenium tablets begin to be largely consumed as a diet integrator, we have administered 100 micrograms/day of selenium as selenium methionine to 8 euthyroid female subjects with a positive iodine-perchlorate discharge test who had a previous episode of subacute or postpartum thyroiditis. We have studied subjects with positive iodine-perchlorate discharge test since the test indicates the existence of a subtle defect of thyroid hormone synthesis and therefore these subjects are prone to develop thyroid dysfunction. In contrast to previous findings in hypothyroid children at low iodine and selenium dietary intake, the supplementation of selenium did not decompensate thyroid hormone synthesis of euthyroid subjects with reduced thyroid iodine organification. The lack of any effect of selenium on thyroid hormone synthesis even in subjects with subtle thyroid hormone synthesis defect may be due to the fact that these subjects had a sufficient selenium dietary intake before selenium supplementation and an only marginally reduced dietary iodine intake. PMID- 8227977 TI - Glycosylated hemoglobin in fractions of erythrocytes of different ages. AB - In 65 children and adolescents with type I diabetes and in 23 healthy controls, glycosylated hemoglobin (HbA1) was measured by ion exchange chromatography in a specimen of whole blood and in three fractions of erythrocytes of different ages. The red blood cells were separated in a density-gradient medium. Three layers were obtained. The top one consisted of younger and lighter cells, while the heaviest and oldest cells were found in the bottom layer. The separation was evaluated by measuring the potassium concentration, which was significantly different in each fraction (p < 0.001). In all healthy controls the lowest HbA1 value was measured in the youngest and the highest in the oldest erythrocytes. In the diabetic patients, however, the HbA1 levels in individual red cell fractions depended on the degree of metabolic control during the life span of the cells. Significant correlation was found between the HbA1 levels in the youngest erythrocytes and metabolic control in the two-week period before investigation (p < 0.001), between the HbA1 content of the medium-aged cell fraction and control two months before assay (p < 0.001), and between HbA1 levels in the oldest erythrocytes and control three and especially four months prior to investigation (p < 0.001). The results of HbA1 determination in the fraction of the youngest cells which show the highest correlation (-0.75) with the glucosuria two weeks prior to assay indicate to be one of possibilities to get information about metabolic control in that period. PMID- 8227978 TI - Evaluation of serum total thyroxine and triiodothyronine and their serum fractions in nonthyroidal illness secondary to congenital heart disease. Studies before and after surgery. AB - Serum thyroid hormones, serum thyroxine-binding proteins and serum thyroid hormone fractions have been measured in children with congenital heart disease before and after open cardiac surgery. Twenty prepubertal patients, mean (+/- SD) age 3.6 +/- 3.7 yr, were studied before, immediately after, and 24 and 48 h after surgery. A control group of 6 normal prepubertal children was also studied in basal conditions. Serum TSH was normal in all samples collected. Significantly low mean levels of serum TBG (261 +/- 57 vs 456 +/- 71 nmol/L in normals), serum TBPA (2692 +/- 1119 vs 5999 +/- 2226 nmol/L), serum TBG-bound T4, serum TBPA bound T4, serum TT3, serum TBG-bound T3 and free T3 were found before cardiac surgery in the patients. While serum binding proteins did not change after surgery, significant decrements in serum TT4, serum TBG-bound T4, serum TT3, serum TBG-bound T3, serum albumin-bound T3 and free T3 were observed after surgery. Free T4 and albumin-bound T4 remained normal. Our study shows that many features of nonthyroidal illness were present in our patients before surgery. In this context, the stress of surgery induced further alterations in several parameters of thyroid metabolism. It is concluded that the changes occurring in this model of chronic, as well as acute, nonthyroidal illness reflect adaptative changes, rather than altered thyroid function, as shown by normal serum free T4, serum albumin-bound T4 and serum TSH. PMID- 8227979 TI - Effect of insulin treatment on osteocalcin levels in diabetic children and adolescents. AB - To investigate bone mineral metabolism in insulin-dependent diabetes mellitus, serum osteocalcin, a marker of bone formation, was measured in 31 diabetic children at onset of disease and 15 days after metabolic improvement by insulin therapy. As a control group for osteocalcin levels we studied 31 healthy sex- and age-matched children. Mean values of serum osteocalcin at onset of diabetes were significantly lower than in control group (p < 0.001), but we did not find any difference after 15 days of insulin therapy. Osteocalcin and parathyroid hormone concentrations were significantly greater after 15 days of insulin treatment than at onset of disease (p < 0.001 and p < 0.01, respectively). The osteocalcin levels were negatively correlated both with fructosamine and with glycosylated hemoglobin (p < 0.01 and p < 0.001, respectively), and positively correlated with the degree of metabolic acidosis at onset (p < 0.05). Therefore we postulate that during glycometabolic imbalance there is a decrease in bone turnover that could be one of the etiological factors of diabetic osteopenia. PMID- 8227980 TI - Measurement of cAMP accumulation in Chinese hamster ovary cells transfected with the recombinant human TSH receptor (CHO-R): a new bioassay for human thyrotropin. AB - Circulating TSH bioactivity may vary in several clinical and experimental conditions. Since the reliability of the current methods for the measurement of TSH bioactivity is limited, a new bioassay based on cAMP accumulation in Chinese Hamster Ovary cells transfected with recombinant human TSH receptor (CHO-R) was set up. The sensitivity was 0.3 +/- 0.1, 0.4 +/- 0.1 and 0.01 +/- 0.01 micrograms/L for TSH IRP 80/558, recombinant human TSH and bovine TSH, respectively. Standard curves were parallel, and the intra- and inter-assay coefficients of variation were 13 +/- 1.1% and 22 +/- 1.9%, respectively. LH, FSH, CG and TSH subunits did not stimulate cAMP accumulation up to high concentrations. Circulating TSH was partially purified by immunoaffinity separation and concentrated before being bioassayed. However, plain sera with high TSH levels, such as those from primary hypothyroid patients (PH), could be directly tested in CHO-R bioassay, provided that sera were added at concentrations lower than 10%. TSH from 6 normal subjects had biological to immunological ratio (B/I) ranging from 0.6 to 2.1 (mean +/- SD = 1.4 +/- 0.5). TSH from 6 patients with PH showed bioactivity significantly lower than in normals (B/I = 0.6 +/- 0.3; p < 0.001; range = 0.3-1.1). TSH from 5 patients with central hypothyroidism of hypothalamic origin (CH) had undetectable basal bioactivity (B/I < 0.2), which normalized in only one patient after acute TRH and in all patients after chronic TRH administration. In conclusion, CHO-R cells provide an excellent tool for evaluating TSH bioactivity, owing to high sensitivity, specificity, reproducibility and feasibility of the assay.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227981 TI - Low dose orally administered arginine is able to enhance both basal and growth hormone-releasing hormone-induced growth hormone secretion in normal short children. AB - Aim of this study was to verify whether arginine (ARG), which likely inhibits hypothalamic somatostatin release, has an enhancing effect on the GHRH-induced GH rise, even when administered orally at low dose. To this goal we studied the effects of 4 g orally administered ARG, either hydrochloride (ARG-H) or aspartate (ARG-A), on both basal and GHRH (1 microgram/Kg i.v.)-stimulated GH secretion in 31 children with familial short stature (11 males and 20 females, aged 5.5-13.8 yr, pubertal stage I-III, and compared the results with those of i.v. infusion of 0.5 g/kg ARG-H. Oral ARG-H (Group A, n = 11) induced a significant increase of basal GH levels (4.2 +/- 1.3 vs 1.0 +/- 0.4 micrograms/L, p < 0.02) and enhanced the GH response to GHRH (41.1 +/- 8.6 vs 25.3 +/- 6.7 micrograms/L, p < 0.02). Oral ARG-A (Group B, n = 10) induced a slight, but not statistically significant increase in serum GH levels (3.4 +/- 1.5 vs 1.0 +/- 0.3 micrograms/L) and enhanced the GHRH-induced GH rise (49.7 +/- 9.8 vs 26.1 +/- 8.4 micrograms/L, p < 0.05). Intravenous ARG-H (Group C, n = 10) stimulated basal GH levels (6.2 +/- 1.2 vs 1.2 +/- 0.3 micrograms/L, p < 0.005) and increased the GHRH-induced GH rise (46.7 +/- 5.0 vs 17.1 +/- 2.3 micrograms/L, p < 0.005). This response was similar to those after oral ARG-H or ARG-A plus GHRH. No variation was observed in PRL levels after oral ARG (either ARG-H or ARG-A) and/or GHRH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8227982 TI - Daily variations in catecholamine excretion are not influenced by very low calorie diet in obese women. AB - The increase in metabolic efficiency during energy restriction seems to be an established phenomenon in obese patients. The sympathoadrenal system is involved in the control of energy expenditure and the catecholamine responses to stimuli vary during the day. We therefore studied the circadian pattern of urinary catecholamine excretion during 4-h collections for two consecutive days in a group of 20 obese female patients during and after a very low calorie diet (500 kcal/die). The diet period induced a significant weight loss in all the patients studied (99.1 +/- 3.7 vs 92.5 +/- 4.1 Kg; p < 0.01). The mean daily excretion of epinephrine did not change after 24 days of diet restriction when compared with the value obtained at the 4th day (12.0 +/- 2.5 vs 10.3 +/- 2.2 nmol/4 h respectively; p = NS) while a slight decrease was observed in the mean daily excretion of norepinephrine (52.5 +/- 8.7 vs 66.6 +/- 9.3 nmol/4 h respectively; p = NS). A circadian rhythm was detected for epinephrine and norepinephrine excretion both during and after very low calorie diet. No significant changes were found in the chronobiological characteristics of epinephrine and norepinephrine with the peak of excretion in the afternoon both during (epinephrine: 16:30 h; norepinephrine: 16:45 h) and after the diet (epinephrine: 17:35 h; norepinephrine: 18:00 h). It seems doubtful that alterations in the chronobiological pattern of catecholamines play a role in the metabolic adaptation occurring during very low calorie diet in obese females. PMID- 8227983 TI - Muscle autoantigens in thyroid associated ophthalmopathy: the limits of molecular genetics. AB - Unlike autoimmune thyroid disease (AITD) in which a number of autoantigens have been identified and characterized, the situation in thyroid associated ophthalmopathy (TAO) is far from clear. A number of candidate antigens have been identified by probing Western blots of orbital tissue (OT) with sera from TAO patients, the most frequently cited being proteins of molecular weight 23, 28, 55, 64, 78 and 120 kilodaltons. In an attempt to identify autoantigens in TAO we have produced a lambda gt11 human eye muscle expression library. This has been screened with sera from four patients with severe TAO whose antibodies bind to one or more of the aforementioned candidate antigens or to a thyroglobulin/acetylcholinesterase (Tg/Ache) shared epitope. Four clones were isolated and characterized; clone R14 encodes the carboxyl terminal 193 amino acids of an IgE binding protein, clones R10 and R13 encode unknown proteins having significant similarity with heat shock protein 27 and the U1 small nuclear ribonucleoprotein respectively. Clone R1 encodes an unknown peptide of 347 amino acids having no similarity with proteins in available data banks. R1 clone affinity purified autoantibodies bind to a protein of Mr 78 kD in a Western blot of porcine eye muscle tissue. Autoantibodies to the R1 recombinant lysogen were clearly demonstrated in 5 of 20 sera from Graves disease patients, its role merits further investigation. The possible relevance of these clones to the pathogenesis of TAO is discussed as well as the limitations of this type of approach in the identification of unknown autoantigens. PMID- 8227984 TI - Octreotide treatment does not affect the size of most non-functioning pituitary adenomas. AB - The somatostatin analogue, octreotide (OC) has commonly been used in the management of growth hormone- and thyrotropin-secreting pituitary tumors, and shown to be effective both on hormone production and tumor size. Because OC receptors may be expressed also in some nonfunctioning pituitary adenomas, it has been postulated that OC might play a role in the treatment of these tumors as well. In the present study, the morphological effects of OC administration, as assessed by computer tomography (CT) scan, were evaluated in 8 patients (5 men, 3 women, age range 25-79 yr) affected by non-functioning pituitary tumors. The drug was given sc at the dose of 100 micrograms tid for 3-6 months. No significant change in visual field or tumor size occurred after OC treatment in 7 patients, whereas one showed a significant improvement of visual field associated with a decreased tumoral mass. These data suggest that OC is not an effective drug in the management of nonfunctioning pituitary adenomas. PMID- 8227985 TI - Androgenic granulosa cell tumor of the ovary: in vivo hormonal studies. AB - Androgenic granulosa cell tumors of the ovary are rare and have not been well studied hormonally. A 20-year-old woman complaining of secondary amenorrhea with high plasma testosterone (295 ng/dl) and a right ovarian tumor was studied. Plasma testosterone levels decreased after two days of dexamethasone (36 ng/dl), increased after hCG administration (538 ng/dl) and a unilateral right ovarian gradient of testosterone was noted at the selective catheterization of ovarian veins. After removal of the granulosa cell tumor, testosterone concentration returned to normal (68 ng/dl) and the patient had resumption of normal menses. PMID- 8227986 TI - Lymphocytes as a source of hormones and peptides. PMID- 8227987 TI - Multiple mitochondrial DNA deletions associated with age in skeletal muscle of rhesus monkeys. AB - We have examined skeletal muscle for the presence of age-associated mitochondrial DNA (mtDNA) deletions from 16 rhesus monkeys (age range 6-27 years). All animals over 13 years of age contained potential mtDNA deletions, whereas the presence of deletions was greatly reduced or absent in younger animals. The specific deletion patterns varied from individual to individual. Numerous mtDNA deletions accumulate with age in skeletal muscle from a nonhuman primate, indicating that the rhesus monkey may provide an excellent animal model to study mtDNA deletions. Further, the existence of multiple mtDNA deletions supports the possibility that they may contribute to geriatric muscular deficits, which are nearly universal in occurrence yet poorly understood. PMID- 8227988 TI - Effect of age and high sucrose diet on 2-deoxyglucose uptake in perfused hindlimb. AB - The purpose of this investigation was to evaluate insulin-stimulated skeletal muscle glucose uptake in male Fischer 344 rats, age 6, 12, and 27 mo, fed either a sucrose (S: 66.6% sucrose, 17.6% protein, 6.4% fat) or sucrose-free (SF: 66.6% starch, 17.6% protein, 6.4% fat) diet for 3 mo. Skeletal muscle glucose uptake (Rg) in perfused hindlimbs was estimated from the uptake and subsequent phosphorylation of radiolabeled 2-deoxyglucose (2DG) in the gastrocnemius (GN), extensor digitorum longus (EDL), and soleus (SOL) muscles. Rat hindlimbs were perfused at a rate of 10 ml.min-1 with a modified Krebs Henseleit buffer containing bovine red blood cells (hematocrit: 40%) and 5.85 mmole.L-1 glucose along with 358 pmoles.L-1 followed by 3580 pmoles.L-1 insulin. There was no effect of diet on plasma glucose levels measured at weeks 1, 7, and 11 of the dietary period. A significant effect of age on estimated glucose uptake in the GN was demonstrated due primarily to greater uptake in the 27-mo compared to the 6 mo-old animals. This significant effect of age was not evident in the EDL or SOL, nor were there significant effects of diet in any muscle. These data suggest that insulin-stimulated glucose uptake in perfused hindlimbs is not attenuated with senescence or with the feeding of a sucrose diet for 3 months. PMID- 8227989 TI - Pathological characterization of male Wistar rats from the gerontology research center. AB - Male Wistar rats aged 6-26 months were obtained from the colony of The Gerontology Research Center of the National Institute on Aging, and pathological profiles were assessed. One hundred animals were sacrificed at 6, 12, 18, 21, 24, and 26 months and used for cross-sectional determinations; also, 150 animals were followed longitudinally and sacrificed when clinical signs of moribundity appeared. Renal disease contributed the most common pathology observed in both studies (found in over 70% of the animals examined), with neoplasms a secondary problem (pituitary tumors were by far the most prevalent, with adenomas present in approximately 20% of the animals). This analysis represents the first complete pathological characterization of this commonly used rat model for aging research, and offers an opportunity for comparison with other rat strains. PMID- 8227990 TI - Specific humoral immunity in the elderly: in vivo and in vitro response to vaccination. AB - Vaccinations are often unsuccessful in preventing infection among elderly populations because of generally poor humoral immune responses. We have used tetanus toxoid (TT) antigen to stimulate in vitro anti-tetanus toxoid antibody (anti-TT) synthesis and have found that lymphocytes from many healthy elderly individuals have a reduced production of anti-TT in vitro compared to young adults. This is associated with decreased numbers of B cells secreting anti-TT IgG and a decrease in the mean amount of anti-TT IgG produced per TT-specific B cell. In the present study we report that immunization results in a significant increase in serum titers in young adults for up to one year, whereas levels in old adults fall to baseline by 6 months. The number of B cells that secrete anti TT IgG increases after immunizations in both young and old subjects, although the number in old subjects is significantly lower than in young subjects at all times except 6 months after. The mean amount of anti-TT produced per B cell (B-cell potency) is significantly lower for the old adults both before and at 6 and 12 months after booster. Immunization does not significantly change the mean amount of anti-TT produced per B cell for either age group. The decreased response to immunization with aging is associated with decreased numbers of specific Ab secreting B cells and usually decreased potency of those B cells. PMID- 8227991 TI - Longevity is moderately heritable in a sample of Danish twins born 1870-1880. AB - The heritability of human longevity was investigated in a sample of 218 pairs of monozygotic (MZ) and 382 pairs of like-sex dizygotic (DZ) Danish twin pairs born 1870-1880. Twin similarity for age at death was significant for MZ twins but nonsignificant for DZ twins. The heritability (h2) of life span estimated from the best-fitting biometrical model was statistically significant but moderate in magnitude (h2 = .333 +/- .058). Heritability of longevity did not vary by gender, and the pattern of twin resemblance was more consistent with nonadditive as compared to additive genetic effects. In addition, evidence for a genetic association between premature and senescent deaths was observed. Although environmental factors accounted for a majority of the variance in life span, the relevant environmental factors appeared to be those that create differences rather than similarities among reared-together relatives. Findings are discussed in terms of their relevance for understanding the inheritance and evolution of human life span. PMID- 8227992 TI - Aging and insulin sensitivity: role of blood pressure and sympathetic nervous system activity. AB - BACKGROUND: The purpose of this study was to determine whether age is a predictor of sensitivity to the peripheral effects of insulin on carbohydrate metabolism independent of the potential influences of the level of sympathetic nervous system (SNS) activity and blood pressure (BP). METHODS: In 60 human subjects (age range 19-78 years), insulin sensitivity, SI, was determined from glucose and insulin levels obtained during an intravenous glucose tolerance test, and plasma norepinephrine (NE) levels were measured to estimate SNS activity. RESULTS: There were age-associated increases in plasma NE level (r = .585; p < .001) and mean arterial BP (r = .516; p < .001), and an age-associated decline in SI (r = -.352; p = .04). However, in stepwise multiple regression analysis, body mass index (BMI) and mean arterial BP were the only independent predictors of SI, accounting for 43% of the variance in SI; age, plasma NE level, plasma epinephrine level, and fasting plasma glucose did not enter the model. Although these results suggest an age-associated decline in insulin sensitivity, this decline appears to be associated with BMI and mean arterial BP rather than aging per se. In addition, the age-associated increase in SNS activity was not found to be an independent predictor of insulin sensitivity. CONCLUSION: This study demonstrates that in addition to BMI, blood pressure should be considered as another confounding factor in studies of insulin sensitivity in human aging. PMID- 8227993 TI - Body composition study in the elderly: comparison between tritium dilution method and dual photon absorptiometry. AB - BACKGROUND: The methods commonly employed in body composition study in elderly persons derive fat-free mass (FFM) by measures of total body water (TBW); these methods assume FFM hydration constant (TBW/FFM = 0.732). The aim of our study was to verify the accuracy of the TBW method in FFM estimation and to study the variability of fat-free mass hydration (TBW/FFM) in healthy elderly subjects. METHODS: We assessed FFM in 27 healthy old subjects (76 +/- 7 yrs) by dual photon absorptiometry (DPA) and by TBW measured by 3H2O. Furthermore, to verify the accuracy in FFM estimation by our methods, we measured resting metabolic rate (RMR) by indirect calorimetry. RESULTS: FFM hydration was 71.9 +/- 4.9 (range 63.6-80.4%), a range larger than that reported by direct chemical analysis in adults. The FFM hydration measured in our subjects was not different from the 73.2 value currently used in healthy adults. FFM values derived by TBW (assuming TBW/FFM = 0.732) were not significantly different from FFM measured by DPA. We correlated FFM with resting metabolic rate and found a similar correlation between RMR and FFM-DPA (r = .89, p < .001) and FFM-TBW (r = .85, p < .001). CONCLUSIONS: Both methods employed in our study have proved to be accurate in estimating FFM in elderly persons; particularly, the value of 0.732 assigned to FFM hydration to derive fat-free mass in adults can be employed also in body composition study of the elderly. PMID- 8227994 TI - Body mass index and fat patterning as correlates of lipids and hypertension in an elderly, biracial population. AB - BACKGROUND: Associations between anthropometric measurements of fat patterning and risk factors for cardiovascular disease have been demonstrated in several populations composed predominantly of White, young or middle-aged adults. Fat pattern changes with aging and older adults tend to have a more central or abdominal fat distribution. Few investigators have focused on relationships of fat patterning with plasma lipids or blood pressure in older adults or in Blacks. METHODS: Body mass index (BMI), fat patterning, plasma lipids, and blood pressure were examined in participants of the Charleston Heart Study. Subjects included 216 White men, 95 Black men, 320 White women, and 155 Black women with a mean age of 72.7 years (range 61-106). RESULTS: In each of the four race and gender groups, BMI was inversely correlated with HDL cholesterol and positively correlated with hypertension. BMI tended to be positively associated with total cholesterol, although the relationships were not statistically significant. After controlling for the effects of BMI, age, smoking, and alcohol intake (using regression analysis), waist to hip ratio was associated with cholesterol levels in the White men and women and the Black men. In similar models, waist circumference in Black women was inversely associated with HDL. There were no significant relationships between waist to hip ratio and hypertension in any of the groups after controlling for BMI. CONCLUSION: In an elderly cohort, waist to hip ratio was associated with plasma lipids, but not with hypertension, when the effects of BMI were held constant. PMID- 8227995 TI - Psychopathology associated with Alzheimer's disease and related disorders. AB - BACKGROUND: This study compared the occurrence of psychiatric symptoms in a large group of community-residing participants enrolled in an Alzheimer's disease patient registry who met clinical diagnostic criteria for Alzheimer's disease or multi-infarct dementia, as well as mixed Alzheimer's disease and multi-infarct dementia. METHODS: Psychiatric morbidity observed from a psychiatric examination of 514 patients with Alzheimer's disease, 135 patients with multi-infarct dementia, and 86 cases with mixed dementia was analyzed using analysis of variance comparing symptoms across groups and chi-square test for differences in frequency of occurrence. Associations between pairs of symptoms were also reported. RESULTS: The frequency of occurrence and patterns of psychiatric problems were similar in patients with Alzheimer's disease and multi-infarct dementia. Agitation was the most frequent symptom, followed by depression, apathy, and behavioral disorders. Patients with mixed dementia had significantly more psychopathology. CONCLUSION: This study demonstrated that significant psychopathology occurs in individuals with Alzheimer's disease or multi-infarct dementia. The high levels of symptoms in mixed dementia suggest that the two conditions have a synergistic effect on behavioral problems. The prevalence of multiple symptoms is higher than previously reported. PMID- 8227996 TI - The relationship of visual and hearing impairments to disability: an analysis using the longitudinal study of aging. AB - BACKGROUND: Little is known about the relationships of visual impairment and hearing impairments to physical disability. The purpose of this work is to determine if persons 70 years of age and over with these impairments are at risk for increased disability in basic physical activities of daily living (ADLs) compared to persons without these impairments. METHODS: We used as our data source the baseline (1984) and the 1988 reinterview from the Longitudinal Study of Aging, a nationally representative survey of noninstitutionalized persons 70 years of age and older. To determine the relationships of visual impairment and hearing impairment to future four-year disability, we used multiple variable modeling, controlling for demographic variables, selected chronic conditions, and baseline disability. RESULTS: Persons with visual impairment were 1.37 (95% CI:1.20-1.57) times more likely to have increased disability in ADLs than those without visual impairment. Hearing impairment was not independently related to increased ADL disability. CONCLUSIONS: Visual impairment by itself is an independent risk factor for future ADL disability. In light of the enlarging older population, maneuvers to ameliorate visual impairment may help to minimize the increase in numbers of disabled persons. PMID- 8227997 TI - Risk factors and consequences of bacteriuria in non-catheterized nursing home residents. AB - BACKGROUND: There has been disagreement over the significance of bacteriuria in nursing home residents. METHODS: During an 18-month period, the risks and consequences of bacteriuria (BU) in 195 residents of a skilled nursing facility without indwelling catheters were examined. Clinical and epidemiologic data and urine for culture were collected every 2 weeks to identify risk factors, symptoms, and occurrences of BU. A mean of 23 cultures per resident was collected. RESULTS: Forty-three percent of the study population (35% of men; 47% of women) had "persistent BU" defined as > 10(5) cfu/ml of urine on > 25% of an individual's collected cultures. Women with persistent BU more frequently were incontinent of bowel and bladder (OR 5.3, 6.3, respectively), more likely to be functionally disabled (OR 3.2), to carry a diagnosis of dementia (OR 2.4), and less likely to have suffered a stroke (OR 0.40). Cancer (OR 6.5) was the only risk factor for persistent BU in men. The number of antibiotic courses prescribed, frequency of hospitalizations, and mortality rates were not significantly different between the two BU groups in either men or women. CONCLUSIONS: Persistent BU is common in nursing home residents. The association of bowel and bladder incontinence and functional disability with persistent bacteriuria suggests that treatment or prevention of these risk factors may prevent or decrease the incidence of bacteriuria. There was no evidence of significant adverse outcomes resulting directly from the bacteriuric state. Higher mortality in the bacteriuric group was the result of underlying functional debility and severity of illness rather than the presence or persistence of BU. PMID- 8227998 TI - Attentional blocks are not responsible for age-related slowing. AB - Reaction time (RT) data in two tasks from a total of 784 adults between 18 and 87 years of age were analyzed to determine the relation between age and parameters of the intra-individual RT distribution. Although the absolute magnitude of the age differences was greatest for the slowest RTs in each individual's RT distribution, there was little or no independent age-related variance in the slowest RTs after controlling for the variance in the fastest RTs. Furthermore, the relation between RT and measures of motor speed, perceptual speed, working memory, and accuracy in several cognitive tasks was of nearly the same magnitude when only the fastest responses were considered as when both fast and slow responses were considered. These results imply that age-related slowing is associated with a shift (and expansion) in the entire RT distribution, and is not attributable to a selective influence on the individual's slowest responses. PMID- 8227999 TI - Are age and education independent correlates of the Mini-Mental State Exam performance of community-dwelling elderly? AB - We examined the effects of selected health conditions and sensory functions, socioeconomic status, age, and education on cognitive functioning in 3,974 community-dwelling individuals aged 65-84 years. Logistic regression analysis was used to examine the independent and joint effects of these variables on borderline (Mini-Mental State Exam [MMSE] of 22-25) and poor (MMSE of < or = 21) functioning relative to adequate functioning (MMSE of 26-30). The effect of age and of education on MMSE performance was relatively stable, even after adjusting for age- and education-related health conditions and sensory impairments that also influenced level of cognitive functioning. These conditions included poor vision, Parkinson's disease, diabetes, depression, stroke (in 65-74-year-olds), and low socioeconomic status (in 75-84-year-olds). Education did not modify the effect of these variables on MMSE performance. Additional studies elucidating further the mechanisms that relate these sociodemographic factors to cognitive performance are warranted, as are studies of the relationship between these factors and the incidence of cognitive impairment. PMID- 8228000 TI - The influence of gender and age on Halstead-Reitan neuropsychological test performance. AB - Age-cohort differences for seven tests from the Halstead-Reitan Neuropsychological Battery, for overall performance on the Impairment Index, and for a multivariate composite of the seven scores, were examined for 427 subjects falling into six age groups (15-24, 25-34, 35-44, 45-54, 55-64, and 65 years and over). Univariate and multivariate-discriminant analyses were performed. Poorer performance was observed across age-cohorts (linear trend) for all tests and the Impairment Index. A very modest Age-Cohort (linear trend) x Gender interaction was observed only for the Category Test. Decline in Category Test performance across age-cohorts was more pronounced for women than for men. For women, the best discrimination among age groups was observed for the Category Test. For men, Tactile Performance Localization Test discriminated best. The poorest discrimination among age groups was seen for Finger-Tapping for men and Tactile Performance Test--Total (time score) for women. Results were consistent with a model of specific nested within general decline. Although linear-trend across age groups was observed for all tests, the strongest discriminations were associated with neuropsychological tests of abstraction and complex problem-solving performance. PMID- 8228001 TI - Stress, social support, and psychological distress among the Chinese elderly. AB - This study examined the relationships among stress, social support, and depressive symptoms with data provided by a large representative sample (N = 2,721) of older adults who live in the People's Republic of China. Many Western investigators believe that elderly people respond to stressful experiences by mobilizing assistance from their significant others. However, recent American studies suggest that this may not always be true, and that when certain stressors arise, older adults may actually receive less support from significant others. The purpose of our study was to evaluate these competing views in a culturally diverse setting. The findings reveal that both perspectives have something to offer and that financial strain (i.e., one kind of stressor) may increase some kinds of assistance (i.e., economic support) at the same time that it erodes other types of support (e.g., emotional support). PMID- 8228002 TI - The changing concentration of the older nonmetropolitan population, 1960-90. AB - Changes in the absolute and relative size of the elderly population since 1960 are decomposed into the underlying demographic components for metropolitan and nonmetropolitan areas and for subregions of the United States. Specifically, we examine the components of net migration and natural increase for those aged 0-64 and those 65 or older. Generally, the natural increase component for those 65 and over has increased since 1960, whereas that for those under 65 has declined. Metropolitan areas have consistently lost, and nonmetropolitan areas gained elderly migrants. Trends in elderly population change are far from uniform across nonmetropolitan America. In general, the "aging" of the nonmetropolitan population was predominantly due to elderly migration during the 1970-80 decade, and to the loss of young people both before and afterward. Recent trends give little support for the view that the 1970s was the beginning of a new phase of deconcentrated settlement, even for elderly persons. PMID- 8228003 TI - Age differences in self-assessments of health: age changes, cohort differences, or survivorship? AB - Do older people tend to exaggerate their health problems? Or do they downplay them? Do such tendencies change as people age? Are they a function of cohort membership? Or are differences in health perceptions among elderly people due to differences in survivorship within the elderly cohorts? These questions are examined with longitudinal data from the Yale Health and Aging Project, a stratified probability sample of elderly persons in New Haven, Connecticut. Complete health assessments are available for 2,583 community residents 65 and older in 1982, and complete follow-up date are available for 1,319 respondents in 1988. Multivariate regressions of self-assessed health on concurrent 1982 and 1988 objective health status and longitudinal analysis of residual values from these equations show that: (a) older survey respondents gave disproportionately positive health assessments, and (b) processes of aging, selective survivorship, and cohort differences all appear to play a role in creating this pattern. PMID- 8228004 TI - Differences between nursing home admissions and residents. AB - The use of a cross-sectional sample of nursing home residents rather than a sample of admissions to estimate admission characteristics carries a potential bias. The purpose of this study was to fill this void by comparing abstracted records data for an admissions cohort (n = 1,118) and a residents cohort (n = 830) residing in the same nursing homes. Compared to residents, admissions were significantly more dependent in their ability to get around and to dress themselves, received more clinical services, and had a higher rate of medication use. Over a 12-month period, admissions had a fivefold greater likelihood of being discharged to community, but about the same mortality rate as residents. Within both groups, those discharged to the community as well as those who died had expenditures that were almost twice as high as those of their counterparts who remained alive in the nursing home. PMID- 8228005 TI - Principal lifetime occupation and MMSE score in elderly persons. AB - Age and education are known to be independently associated with MMSE score. This study addressed the relationship of principal lifetime occupations, which are related to both age and education, with MMSE scores in 524 community-dwelling elders aged 70 and older. The data suggest that principal lifetime occupations are correlated with MMSE scores independently of the effect of age and education. As an example, formers have an average MMSE score 2.33 points lower than white collar workers in a multiple linear regression analysis (95% C.I. 0.46-4.21), even after adjustment for age, education, and financial dissatisfaction. PMID- 8228006 TI - Concerns regarding adult children's assistance: a comparison of young-old and old old parents. AB - This study describes differences between young-old (ages 60-74) and old-old (ages 75-90) parents' concerns regarding assistance provided by adult children. Old-old parents expressed more concern regarding amounts of help than did younger parents; there were no age differences in levels of concern regarding help outcomes. The percentage of children receiving parents' help and reliance on formal services predicted level of concern regarding help outcomes for the young old parents, but not the old-old parents. Familism values and the extent of help received predicted level of concern regarding help amounts for the old-old parents, but not for younger parents. Implications for interventions to minimize parents' concerns and for further study of family assistance to the older people are discussed. PMID- 8228007 TI - [Comparison of different types of ovarian wound closure in rats. Role of biological glue]. AB - Three techniques for closing the ovary have been compared. The test was carried out on 30 rats' ovaries. After the ovary had been cut with scissors it was closed with rapid acting Tissucol (a biological glue) or closed with interrupted stitches of 10/0 Vicryl, or left to close by itself. Sixty days later the ovaries were looked at macroscopically and histologically. The macroscopic score was established according to the presence of adhesions, the size of the ovary, the presence of cysts; and the histological score was carried out according to the presence of granulomatous macrophage lesions, the degree of fibrosis and the existence of germ cell cysts. The results were identifically the same as far as these five different criteria were concerned. All the same, Tissucol brought about less fibrosis and less atrophy of the ovary. Tissucol, therefore, is a good alternative for suturing the ovary as compared with stitching or no formal closure after the removal of ovarian cysts, particularly laparoscopically. PMID- 8228008 TI - [Severe oligospermia with normal FSH. Comparison between radioimmunologically and immunoradiometrically determined levels of LH]. AB - We have noted in a group of oligospermic patients with normal FSH levels and normal sized testes that there can be a significant drop in the LH levels estimated by IRMA, compared with a control group of fertile men. On the other hand, there is no correlation between IRMA and RIA levels of LH in oligospermic men when there is such a correlation in the control group. As oligospermia is usually testicular in origin preliminary results suggest that there may be a disturbance of gonadotrophin secretion in one type of oligospermia and we intend to continue this preliminary study. PMID- 8228009 TI - [Pregnancy by micromanipulation in a case of excretory azoospermia associated with a natural spermatocele]. AB - Microinjection of capacitated sperm into the perivitelline space of oocytes has been performed in a case of male infertility attributed to excretory azoospermia. In this particular case, a spermatocele detected five years ago was observed, which provide motile spermatozoa. These sperm collected by direct puncture into the cyst, were capacitated using discontinuous Percoll gradient. After ovarian stimulation, seventeen oocytes were collected. Nine of them were used for classical IVF whereas the other eight were submitted to microinjection. Only one oocyte of the second group fertilized and cleaved 48 hours later. A clinical pregnancy was achieved after embryo transfer and a normal, healthy boy delivered at term. PMID- 8228010 TI - [Cancer of the cervix, papillomavirus, contraception and tobacco]. AB - This survey started in May 1989 and deals with 2,436 patients treated by 10 gynecologists from the Brussels region. In addition to routine cytological analysis in cervical smears, an additional test for Human Papilloma Virus (HPV 16, 18 and 33) was advised for these women. Gynecologists filled in a questionnaire about these patients. In cases with benign cytology, virus prevalence was 2 times greater in woman who took contraceptive pills, over at least 6 months, than in each of the other groups using either diaphragm or male condoms or no contraception, or at menopausal age. It was strange to find that viral prevalence varied according to the pill trade mark, but not according to the pill hormonal content. No confounding factor in population recruitment could be detected. In contrast to viral infection, prevalence of cancer lesions was not increased in the oral contraception group. Squamous intraepithelial lesions of Low Grade and High Grade (SIL LG and HG) were on the contrary 2 times less frequent in women taking the pill, with or without HPV infection. During follow up of 223 women, the same HPV genotype persisted in SIL lesions but it disappeared within 4 to 8 weeks in 68% of normal cervical smears. However, beyond this period, the proportion of women with persisting virus remained nearly constant, indicating that a subgroup may be less able to mount a defence against the infection. Risk factor of cigarette smoking for SIL LG or HG was 1.6 in women with HPV and 4.0 in those with no virus. This factor was 3.3 in women with oral contraception and 4.5 in the other group. PMID- 8228011 TI - [Value of biological glue in CO2 laser conization. Experience with 32 cases]. AB - The effects of the use of biological glue in the crater that is left after CO2 laser conisation was studied in a series of 32 cases and compared with a previous study carried out using the same technical methods by the same surgeons. The use of biological glue has as its aim to improve capillary haemostasis after conisation; and so increasing the security of leaving these conisation craters in order to improve the chance of normal morphology later. Biological glue has furthermore never given a febrile reaction, nor has it ever increased the likelihood of scar stenosis. Biological glue however is just an extra method of helping normal morphological healing with a good transformation zone and retaining fertility. When conisation is carried most extensively (in particular as far as depth is concerned) and with the use of powerful laser beams, the application of biological glue seem to guarantee most forms of postoperative haemostasis and good healing. PMID- 8228012 TI - [Castleman's disease localized in the uterus or an inflammatory uterine pseudotumor. A case report and review of the literature]. AB - The authors report a case of Castleman's disease localised to the uterus. The treatment was surgical removal of the lesion with conservation of the uterus. Though in localised forms the prognosis is always favourable, multicentered lesions can suggest lymphoma. In our case that we report the anatomo-pathological appearances were not characteristic and this has led the authors to think of a differential diagnosis such as an inflammatory pseudo-tumour of the uterus of which only two cases have been previously reported. PMID- 8228013 TI - [Krukenberg's tumor: still a somber prognosis. 4 cases]. AB - The authors report four cases of Krukenberg's tumour. They point out its rarity and that the diagnosis can come as a surprise on histological examination. The tumour always has Signet ring cells. How the ovary comes to be involved is not known as yet. Treatment is mainly surgical with added chemotherapy but the prognosis is still very poor indeed. PMID- 8228014 TI - [HIV-1 seroprevalence in pregnant women in Libreville (Gabon)]. AB - The authors tried to find out the incidence of HIV positive women in 634 deliveries in the two large maternity units in Libreville. The prevalence was 1.26% which is a net rise compared with previous figures in the same city, but remains lower than figures for other large cities in Central Africa. Transmission is mainly by the sexual route and in particular by the heterosexual route and the virus particularly affects young women at their most fertile which makes the epidemic all the more serious because of congenital aids. PMID- 8228015 TI - [Medullary aplasia during treatment for congenital toxoplasmosis in a twin pregnancy]. AB - The authors report a case of a patient who in the 24th week of a twin pregnancy became sero-positive for toxoplasmosis. This was diagnosed by cordocentesis as being infected, and the treatment was therefore started with pyrimethamine and sulfadiazine and folic acid at the 28th week of pregnancy. At 35 weeks, the patient had an acute medullary aplasia due to the absence of the folates. The mother's state was improved rapidly by giving her folinic acid and the twins were normal haematologically. In this case, the authors point out how important the folates are in a pregnancy, especially in twin pregnancies, and point out the precautions that have to be taken when treatment with pyrimethamine and sulfadiazine is started for congenital toxoplasmosis. PMID- 8228016 TI - [Massive fetomaternal hemorrhage and prevention of fetomaternal Rhesus incompatibility. The failure of our present system of prevention]. AB - We had a case where risk of rhesus D feto-maternal immunisation occurred following failure to diagnose feto-maternal haemorrhage (HFM); and it was shown up by rhesus negative mother with a rhesus positive fetus being diagnosed as having has a massive HFM only three days after delivery. Giving the mother the standard dose of Anti-D immunoglobulin without a previous test to find out how serious the HFM was showed that we do not test for this normally. So it seems to us necessary when considering prophylaxis of rhesus D immunisation to go back to first principles and carry out Kleihauer's test particularly when neonatal anaemia is found in the child. PMID- 8228017 TI - [Jaundice and pregnancy. The role of viral hepatitis]. AB - Through a series of 76 cases of jaundice observed in pregnant women, the authors emphasize on the biggest frequency of viral hepatitis among the all of etiology. In fact that one represents the 2/3 of cases. They insist on the graves forms of the diseases which are responsible of a high maternal mortality and big foetal complications. PMID- 8228018 TI - [Intrahepatic cholestasis in pregnancy. The hepatologist's point of view]. AB - Intra-hepatic cholestasis of pregnancy (ICP) is a specific liver disease that occurs in the third trimester of pregnancy (less frequently in the second trimester) and disappears quickly after delivery. Cholestasis can occur in pregnancy in three situations: a chronic liver disease brought out during pregnancy, intercurrent liver disease or ICP. The serum levels of alanine aminotransferase (ALT) and total bile salts are the most sensitive tests for diagnosing cholestasis in pregnancy. Collaboration between the obstetric team and the liver doctor is needed to find a cause or a factor that increases the risk of cholestasis. Urinary tract infections should always be ruled out. Oral hormonal treatments in pregnancy have not been clearly established as a cause and further investigations are continuing. The maternal prognosis is excellent, but it is important to monitor the prothrombin time and treat any vitamin K deficiency. On the other hand, the fetal prognosis is less good and there is an increase in prematurity and intra-uterine fetal death. When a diagnosis of cholestasis has been confirmed we advise immediately cessation of hormone treatments including natural progesterone. We describe the principals of medical and obstetric management. PMID- 8228019 TI - [The effect of peridural analgesia on uterine contractions]. AB - OBJECTIVE: To study the impact of 0.25% bupivacaine on the frequency and intensity of uterine contractions. METHODOLOGY: 52 patients in labour. Continuous internal tokographic measurement of uterine activity at 10 minutes before instilling the bupivacaine and at 30 minutes after doing so. The true frequency and intensity of the uterine contractions were the two parameters that were studied. RESULTS: There was a significant variation (matched Student's test) between the two parameters that were studied: there was transitory hypokinesia (p = 0.003) and intense prolonged hyperkinesia (p < 0.001). CONCLUSION: 0.25% bupivacaine produces hyperkinetic increases in intensity of the contractions of which the mechanism has not been elucidated; therefore it is inadvisable to add oxytocics in the minutes after instilling bupivacaine. PMID- 8228020 TI - [Delivery after 2 previous cesarean sections. A series of 41 uterine trials]. AB - It has been possible to consider how delivery should be carried out in view of the progress that has been made handling scarred uteruses. Over 21 months 41 tests of uterine function have been authorized in our department out of 67 cases where there were two scars in the uterus (67%). 26 patients delivered vaginally (63.4%) and 5 had Caesarean sections because of failure of the test of the scar. When the relationship between the fetus and the pelvis was satisfactory, the fact that the cervix was not ripe and the presenting part was not engaged, did not prevent carrying out a trial of scar in 78% of cases. It is important to assess conditions continuously during labour and this assessment should include fetal heart monitoring, internal tokometry, fetal pH assessment and ultrasound of the scar at the onset of labour. Oxytocics had to be used in 96.2% of cases because there was at the outset marked dynamic dystocia. Epidural anaesthesia was used in 90.2% of cases. A full obstetric team must be present throughout the whole labour so that the conduct of the labour can be observed, and if necessary corrected quickly if anything in its progress is becoming abnormal. It has become reasonable to carry out tests of uterine scars even after two scars have been made in the uterus because of the absence of any maternal or fetal complications in this series or in the literature. PMID- 8228021 TI - [Diabetes insipidus and postpartum anterior hypophyseal insufficiency]. AB - There have only been thirty cases of total post-partum hypopituitarism published in the literature and these have nearly all been secondary to Sheehan's syndrome. The authors report a case of partial anterior hypopituitarism associated with diabetes insipidus which arose after an uneventful Caesarean operation and the origin of which seems to lie in auto-immune hypophysitis. The authors first describe the morphological and endocrine changes that the hypophysis undergoes during pregnancy and then point out that auto-immune hypophysitis seems to have been only recently recognised. This can be used to explain some cases of post partum hypophyseal insufficiency occurring almost silently without any history of third haemorrhage. Research has been made systematically for anti-hypophyseal antibodies and for specific antibodies of the organ, but has not always been positive. So the diagnosis of auto-immune hypophysitis is often made only after eliminating other reasons for it. A brief review of the physiopathological mechanisms of diabetes insipidus makes it possible to suggest that vasopressinase coming from the placenta together with prostaglandins could play a role. PMID- 8228022 TI - [Diabetes and pregnancy in Africa: a pathology often not recognized]. PMID- 8228023 TI - Why a neuro-oncologist? PMID- 8228024 TI - Brainstem and other malignant gliomas: II. Possible mechanisms of brain infiltration by tumor cells. AB - Gliomas that arise in the brain stem and other malignant gliomas constitute approximately 60% of all brain tumors and have eluded effective therapy, in part because they are able to infiltrate the normal brain. Histopathologic studies have confirmed the presence of infiltrating tumor cells very distant from the glioma mass. We review the neuroimaging and pathologic features of glioma-cell infiltration and some of the complex cellular and biochemical determinants of tumor-cell motility and invasiveness. Understanding how glioma cells become motile and invasive is pivotal to therapeutically targeting the machinery that enables gliomas to infiltrate the brain. PMID- 8228025 TI - Neoplastic meningitis as the presentation of occult primitive neuroectodermal tumors. AB - Seven children and young adults initially presented with subacute meningitis and/or increased intracranial pressure. The diagnosis of neoplastic meningitis secondary to a primitive neuroectodermal neoplasm was delayed by the absence of an obvious primary tumor. The neuroradiologic appearance was that of a basimeningeal infiltrative process, complicated by communicating hydrocephalus or "pseudotumor cerebri." Myelography was important in the diagnosis of disseminated meningeal malignancy in four cases. Cerebrospinal fluid cytologic diagnosis was insensitive but ultimately confirmed in five cases. All seven patients experienced progressive disease despite neuraxis radiotherapy and intensive chemotherapy; six have died. Systemic dissemination to bone and/or peritoneum occurred in three patients while on therapy. In two, a primary parenchymal brain or spinal cord tumor could not be identified at postmortem examination. The presentation of a primitive neuroectodermal tumor as subacute meningitis without an evident primary tumor heralds an aggressive and refractory neoplasm. PMID- 8228026 TI - A reversible case of Kluver-Bucy syndrome in association with shigellosis. AB - The Kluver-Bucy syndrome is characterized by psychic blindness or visual agnosia, blunted affect, hypermetamorphosis, hyperorality, bulimia, and sexual behavioral alterations. To date, there have been fewer than 10 reported cases of Kluver-Bucy syndrome in children, most of them irreversible. We describe here a reversible case of the complete Kluver-Bucy syndrome in a child suffering from Shigella encephalopathy. PMID- 8228027 TI - Neurophysiologic study of beta-thalassemia patients. AB - Neurophysiologic investigations were performed in 34 Chinese patients with beta thalassemia major maintained on long-term desferrioxamine treatment to look for subclinical toxicity in the auditory, visual, peripheral, or central neural pathways. In the auditory pathway study, four patients (12%) had mild sensorineural hearing impairment. Two patients (6%) had increased P 100 latencies in the visual evoked potential study, and nine patients (26%) had abnormal electroretinogram results. All had normal electrooculograms. Ophthalmoscopic examination was abnormal in three patients (9%), and three (9%) had a visual field defect. In the peripheral or central nervous pathways, seven patients (21%) had sensory neuropathy, of which three cases were probably related to diabetes mellitus. All had normal motor conduction velocities. Four patients (12%) had increased cortical latencies of median or posterior tibial somatosensory evoked potential. Abnormalities in multiple neural pathways were seen in four patients (12%). There was a significant association between subclinical toxicity to the peripheral or central nervous systems and serum ferritin level (P < .03) and the presence of diabetes mellitus (P < .002). There was no significant relationship between the age, dosage, or duration of desferrioxamine used and the increased risk of neurotoxicity to the auditory, visual, peripheral, or central nervous systems. There was also no association between the risk of neurotoxicity and the serum zinc, copper, or fructosamine levels. PMID- 8228028 TI - Electrophysiologic correlates of peripheral nervous system maturation in infancy and childhood. AB - Peripheral nervous system maturation in infancy and childhood varies with age, especially during the first 2 years of life. Electrophysiologic values therefore changes significantly between different age groups within these first 2 years and are different from adult values. Normal values of motor and sensory nerve conduction, distal motor latency, F-wave latency, and evoked response amplitude of peripheral nerves commonly tested are reported in 155 healthy children in seven age groups from 1 week to 14 years. Interval changes are clearly shown, and in comparison with adult values, the whole group has significantly slower nerve conduction velocities, with reduced muscle and nerve evoked response amplitudes. These differences are important to recognize when evaluating the peripheral nervous system of children. PMID- 8228029 TI - Attention deficit-hyperactivity disorder and asymmetry of the caudate nucleus. AB - The neurologic basis of attention deficit-hyperactivity disorder (ADHD) is poorly understood. Based on previous studies that have implicated metabolic deficiencies in the caudate-striatal region in ADHD, we employed magnetic resonance imaging to investigate patterns of morphology of the head of the caudate nucleus in normal and ADHD children. In normal children, 72.7% evidenced a left-larger-than-right (L > R) pattern of asymmetry, whereas 63.6% of the ADHD children had the reverse (L < R) pattern of asymmetry of the head of the caudate nucleus. This reversal of normal asymmetry in ADHD children was due to a significantly smaller left caudate nucleus. The reversal in asymmetry of the head of the caudate was most notable in ADHD males. These results suggest that normal (L > R) morphologic asymmetry in the region of the caudate nucleus may be related to asymmetries observed in neurotransmitter systems implicated in ADHD. The behavioral symptoms of ADHD may reflect disinhibition from normal levels of dominant hemispheric control, possibly correlated with deviations in asymmetric caudate-striatal morphology and deficiencies in associated neurotransmitter systems. PMID- 8228030 TI - Attentional disturbance after pediatric closed head injury. AB - The influence of severity of closed head injury and age on attentional functioning was prospectively investigated in 36 children (age range, 7 to 16 years) 6 months after injury. Children were placed into mild, moderate, and severe injury groups using established neurologic criteria. Each child received the Wechsler Intelligence Scale for Children-Revised Digit Span subtest and a continuous performance test. Children with severe closed head injury demonstrated significantly poorer continuous performance test scores than mildly or moderately injured children. Injury severity had no effect on Digit Span scores. Younger children exhibited more pronounced impairment on the continuous performance test relative to uninjured age peers. These results extend the persistence of attentional impairments beyond those of previous reports. Closed head injury is not associated with preferential sparing of sustained attention in younger children 6 months after injury. Brain injury earlier in life may result in delayed vulnerability of information processing skills. PMID- 8228031 TI - Intramedullary astrocytoma presenting as spinal muscular atrophy. AB - We present a 6-year-old patient with a spinal cord tumor who had been followed with the diagnosis of spinal muscular atrophy since the age of 23 months. Reasons for reevaluating the diagnosis of spinal muscular atrophy were the early onset of scoliosis, the slight asymmetry in weakness of extremities, and the appearance of urinary retention in the last 3 days. Magnetic resonance imaging revealed a very long, intramedullary tumor extending from the level of the seventh cervical segment to the conus medullaris, later reported to be a grade I astrocytoma. We therefore recommend that magnetic resonance imaging, a noninvasive and sensitive technique for intraspinal pathologies, be performed in every patient with an atypical form of spinal muscular atrophy. PMID- 8228032 TI - The rapid change to monotherapy with valproic acid in children. AB - A review of hospitalized epilepsy patients at Texas Tech University identified 20 children during a 4-year period who had undergone a rapid change from other anticonvulsants to valproate monotherapy. All patients had difficult-to-treat or intractable epilepsy syndromes. In each case, sodium valproate was introduced over a 2- to 4-day period as all other anticonvulsants were withdrawn quickly and sequentially. No acute side effects were seen during hospitalizations, and only one patient reported gastrointestinal complaints. At time of discharge, 12 of the 20 patients were seizure free. Fifteen of the 20 patients were maintained on monotherapy following discharge. By the end of the study period, 12 of the 20 patients showed improvement with valproate, with nine patients experiencing complete seizure control. A rapid switch to valproate monotherapy can be done safely in an inpatient setting. PMID- 8228033 TI - Myoclonus epilepsy with ragged-red fibers: a clinical and electrophysiologic follow-up study on two sibling cases. AB - We performed a 5-year clinical and electrophysiologic follow-up study on two sibling cases with myoclonus epilepsy with ragged-red fibers. Both had myoclonus, intention tremor, slight muscle weakness, slight mental disturbance, hearing impairment, and optic atrophy. Neither had epileptic attacks or truncal or gait ataxia. Biochemical activity of cytochrome c oxidase was at the lower limit of the normal range of values, and an adenine to guanine transition mutation at nucleotide 8344 in the transfer RNA specific for lysine of mitochondrial DNA was detected in both cases. The electroencephalograms showed slowing of basic patterns, diffuse spike-and-wave complexes, occipital dominant wave-and-spike phantoms, 6- and 14-Hz positive spikes, and photosensitivity. No definite deterioration of basic patterns was seen, and diffuse spike-and-wave complexes and photosensitivity gradually disappeared during the slowly progressive clinical course. P2 latencies of pattern-reversal visual evoked potentials throughout the clinical course and III through V interpeak latencies of auditory brainstem responses at follow-up were prolonged without giant sensory evoked potentials in both cases. PMID- 8228034 TI - Progressive encephalopathy associated with cytomegalovirus infection without immune deficiency. AB - Slowly progressive encephalopathy caused by cytomegalovirus is an unusual disorder, and its pathogenesis remains unknown except for cases associated with the acquired immune deficiency syndrome and organ transplantation. We report a case who showed clinical features of progressive encephalopathy. Cytomegalovirus was repeatedly isolated from urine, and cytomegalovirus-infected cells were detected in bone marrow. Serial computed tomographic head scan revealed periventricular calcification and its progression to the thalamus, cerebellum, and brain stem. On autopsy, there were multiple calcifications and diffuse glial proliferation in the gray and white matter. Perivascular inflammation was only minimal. There was no evidence of immune deficiency. This case suggests that progressive encephalopathy can be caused by cytomegalovirus infection without immune deficiency. This type of cytomegalovirus infection may be unusual, but its serious outcome should remind us to detect it accurately. PMID- 8228035 TI - Value of serum anti-herpes simplex viral IgM antibody testing in empirical antiviral treatment of herpes simplex encephalitis. AB - Early antiviral treatment is important for herpes simplex encephalitis. A reliable test for supporting empirical therapy with antiviral agents is urgently needed. Sixty-six children with fever, focal seizures, and consciousness disturbance had their sera examined for anti-herpes simplex viral immunoglobulin M (IgM) antibody with enzyme-linked immunosorbent assay. The result was positive in all seven patients confirmed to have herpes simplex encephalitis. Only two of the other 59 patients were positive on this test. The average period for the development of serologic positivity was 7.3 days from the onset of neurologic symptoms. Although most sera were not tested in the early stage, all were collected on admission day. Our data suggest that before further evidence indicates another diagnosis, this simple, but specific IgM antibody test could be used as a guide for deciding to continue the antiviral treatment for serologically positive cases or to discontinue antiviral treatment for serologically negative ones. Yet, we would not suggest diagnosing herpes simplex encephalitis with this test alone. PMID- 8228036 TI - Bilateral temporal arachnoid cysts in neurofibromatosis. AB - The occurrence of bilateral temporal arachnoid cysts has been considered as a rare event. Unilateral arachnoid pouches have been reported in a few instances associated with neurofibromatosis. The authors describe a 5-year-old girl with bilateral temporal arachnoid cysts who presented obvious stigmata of van Recklinghausen's disease. To the best of our knowledge, this is the first time that this association has been described in the literature. PMID- 8228037 TI - Epilepsia partialis continua due to cortical dysplasia. AB - We describe a child with progressive neurologic deterioration and epilepsia partialis continua of the right extremities. Magnetic resonance imaging suggested cortical dysplasia of the left frontal region including the central sensorimotor cortex. Subdural grid recordings showed widespread ictal activity and frequent interictal discharges from the motor cortex. A tailored resection was performed, and histologic analysis confirmed cortical dysplasia. This case demonstrates that cortical dysplasia may be the underlying cause in some cases of childhood epilepsia partialis continua of the progressive type. PMID- 8228038 TI - Report of congenital anaplastic astrocytoma discovered in a newborn. AB - Reports of congenital anaplastic astrocytoma have been rare, representing less than 2% of all brain tumors. We present a case of a congenital anaplastic astrocytoma discovered during the first 24 hours after delivery. A recent literature review found 12 additional cases since 1972. The presenting signs, most common findings on neuroimaging, treatment, and outcome are examined. PMID- 8228039 TI - Neurofibromatosis type 1: review of the first 200 patients in an Australian clinic. AB - Neurofibromatosis type 1 is a common multisystem disorder, best managed in a multidisciplinary clinic. In 1991, the first Australian neurofibromatosis clinic was established at the Children's Hospital, Camperdown, and the clinical characteristics of the first 150 families are reviewed. Two hundred individuals were assessed; there was an equal sex distribution, and 55% of cases were sporadic. Advanced paternal age appeared to predispose to new mutations in the neurofibromatosis gene. Cafe-au-lait spots and axillary freckling were important to the diagnosis of neurofibromatosis type 1 during childhood, and neurofibromas and Lisch nodules, although often not appearing until after puberty, were present in almost all patients over 30 years of age. Short stature (27%), macrocephaly (43%), scoliosis (20.5%), and learning disabilities (45%) were common associated features. The prevalence of disease complications was similar to the major US and European studies. PMID- 8228040 TI - Tests for juvenile myasthenia gravis: comparative diagnostic yield and prediction of outcome. AB - The clinical picture, family history, laboratory data, treatment modalities, and outcome in 27 juvenile myasthenics seen over a 25-year period are presented. Onset was after 10 years of age in the majority of patients. Half presented with ocular signs, the other half with generalized-onset myasthenia. Half of those with ocular-onset myasthenia progressed to generalized myasthenia. The female-to male ratio was 3.5:1. Myasthenia gravis was reported in the mother of one patient. Ptosis was the most common presenting sign. It was unilateral at onset in 33% of patients and remained unilateral in 11%. Pharmacologic tests (neostigmine methylsulfate and edrophonium chloride) were positive in 92% of patients. Serology was positive in 63%, whereas repetitive nerve stimulation was positive in 33% when distal nerves were stimulated and in 66% when proximal and distal nerves were stimulated. Seropositivity tended to increase with generalization of the myasthenic process. No statistically significant difference in seropositivity was noted between males and females. Anti-striated muscle antibodies were detected in two patients, neither of whom had thymoma. The yield of repetitive stimulation increased with generalization of the myasthenic process and when proximal nerves were stimulated. No statistically significant difference was noted in the decremental response between seropositive and seronegative patients. The majority of ocular myasthenics were treated with pyridostigmine bromide monotherapy. With generalization of the myasthenic process, other modes of medical and surgical therapies were needed. All patients given corticosteroids ultimately underwent thymectomy. The mean age at presentation and the duration of symptoms at presentation were longer in thymectomized patients than in those without thymectomy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228041 TI - Turner syndrome and occlusion of the internal carotid artery. AB - A 2-year-old girl with Turner syndrome was admitted with left hemiplegia and left facial palsy. Serial cranial computed tomographic scan demonstrated multiple cerebral infarctions in the right putamen and right medial cortical areas. Single photon emission computed tomographic scan revealed hypoperfusion from the right frontal to the right temporal area. Right carotid angiography showed narrowing and occlusion of the right internal carotid artery at the sphenoidal portion. Collateral circulation was not detected between the external and internal carotid arteries. Left carotid angiography revealed that the left anterior artery was narrow, and that the left internal carotid artery provided blood to the right internal carotid artery through the anterior communicating artery. These findings suggested that the cerebrovascular abnormality might be due to congenital hypoplasia of arteries in this patient. The unusual combination of cerebral infarction and Turner syndrome was reported. PMID- 8228042 TI - William Osler: a Philadelphia neurologist. PMID- 8228043 TI - Thoughts on the future of child neurology. PMID- 8228044 TI - Detection of Epstein-Barr virus genome in cerebrospinal fluid from a patient with acquired chorea by the polymerase chain reaction. PMID- 8228045 TI - Perilunate dislocations and fracture-dislocations: a multicenter study. AB - A series of 166 perilunate dislocations and fracture-dislocations from 7 centers was retrospectively studied. The diagnosis was missed initially in 41 cases (25%). A classification system was presented. The perilunate fracture dislocations were more frequent than the perilunate dislocations at a ratio of two to one. The displacement was dorsal in 161 cases (97%) and palmar in only 5 (3%). The dorsal transscaphoid perilunate fracture-dislocations represented 96% of the dorsal perilunate fracture-dislocations and 61% of the whole series. The clinical and radiologic outcome of 115 perilunate dislocations and fracture dislocations with at least 1 year and an average of 6 years 3 months of follow-up was studied. Open injury and delay of treatment had an adverse effect on clinical results, whereas anatomical type had less influence. In cases treated early, the clinical results were satisfactory but the incidence of post-traumatic arthritis was high (56%). In the dorsal perilunate dislocation group of pure ligamentous injuries and in the dorsal transscaphoid group, the best radiologic results were observed after open reduction and internal fixation. In the latter group, the fixation of the scaphoid alone was not always sufficient and left occasionally scapholunate dissociation, lunotriquetral dissociation, ulnar translation of the carpus, or other carpal collapse patterns. The initial appraisal of both the osseous and ligamentous pathology was very important. PMID- 8228046 TI - Scaphoid excision and capitolunate arthrodesis for radioscaphoid arthritis. AB - Eighteen patients with symptomatic radioscaphoid arthritis had scaphoid excision and capitolunate arthrodesis. Eight patients also had a silicone scaphoid replacement. The follow-up period averaged 3 years. Fusion was solid in 12 cases at an average of 8 weeks, and pain was significantly less at follow-up evaluation. Six patients had a pseudarthrosis and five had persistent pain. Immobilization in the pseudarthrosis group averaged 6 weeks, which was significantly less than the group that fused. Two patients underwent successful repeat fusions. Wrist extension averaged 26 degrees, flexion 34 degrees, radial deviation 11 degrees, and ulnar deviation 24 degrees. Grip strength averaged 25 kg. Presence of an implant had no significant effect on motion or strength. Pin track infection and pseudarthrosis were the main complications. Pain relief, functional motion, good strength, and patient satisfaction can be expected after scaphoid excision and solid capitolunate arthrodesis. Kirschner wires should be buried subcutaneously to avoid infection. The addition of a scaphoid implant offered no advantage over simple scaphoid excision. PMID- 8228047 TI - Wrist arthrodesis in post-traumatic arthritis: a comparison of two methods. AB - The Arbeitsgemeinschaft fur Osteosynthesefragen arthrodesis technique is compared with the proximal row carpectomy technique with radiocapitohamate arthrodesis for patients with traumatic and degenerative arthritis of the wrist. Both techniques gave satisfactory functional and x-ray film results in 85% of the patients. Grip strength measured on the Jamar dynamometer at settings 1-5 was 77-83% of the opposite in the dominant extremities and 56-82% of the opposite in the nondominant extremities. Despite the decrease in grip strengths after wrist fusion, grip strength curve patterns were similar for operated and nonoperated extremities. There were no statistically significant differences in grip strength between the two groups. Patients who did not have index and long ray carpometacarpal joints fused had pain at that level requiring a second procedure to arthrodese the joints. Index and long finger carpometacarpal joint arthrodesis at the time of wrist arthrodesis is recommended in patients with heavy labor jobs. PMID- 8228048 TI - Internal fixation of scaphoid injuries using the Herbert screw through a dorsal approach. AB - This prospective study reports on the use of the Herbert screw for fixation of fractures and nonunions of the scaphoid through a dorsal approach. In cases of delayed union and nonunion, screw fixation was combined with bone grafting. Twenty-five patients were treated in this manner, and union rates of 100% for acute fractures and 87% for delayed union and nonunion were achieved. We believe that the dorsal approach provides ready access to the scaphoid, enabling placement of the screw in the best possible position to provide fracture fixation. PMID- 8228049 TI - Revision total wrist arthroplasty. AB - Biaxial design total wrist arthroplasty was used to salvage 13 failed total wrist arthroplasties of various designs. After a follow-up period of 31 months, two wrists had undergone further prosthetic revision for loosening and one wrist was arthrodesed. In the remaining 10 wrists, 8 had no pain, 1 had mild pain, and 1 had moderate pain. Eight patients reported feeling much better, one reported feeling better, and one reported no change. Postoperative range of motion averaged 36 degrees of extension, 19 degrees of flexion, 6 degrees of radial deviation, and 15 degrees of ulnar deviation. X-ray films at follow-up evaluation showed that two revision arthroplasties still in place were loose. The majority of patients had a satisfactory clinical result after revision total wrist arthroplasty with a biaxial design. Loosening of the revision is a significant problem. PMID- 8228050 TI - Kinematics of the wrist and its ligaments. AB - Precise anatomic dissections of unembalmed physiologically intact cadaver specimens were carried out before proceeding with the kinematic investigations on further specimens. Carpal flexor and extensor tendons were used to move the wrist. The analysis of the carpal bone movements was performed according to the finite helical axis motion concept for increments of 10 degrees for each main carpal motion. Separate axes of rotation for each of the bones in the proximal carpal row were found; however, the axes for the lunate and triquetrum bones were close, and the magnitudes of rotation and translation almost equal. One finding was that the axes of rotation of the bones in the proximal carpal row often cross at some particular point. Because the axes of the scaphoid are differently oriented than those of the lunate and triquetrum, shear might occur during wrist motion. This indicates also that the scaphoid and lunate cannot be considered rigidly coupled elements. We also assume that the individual bones of the proximal carpal row self-align themselves as long as they are not constrained by torsion. Several potential mechanisms of flexion motion of the proximal carpal row during radioulnar deviations of the hand were considered. The ligament function is still an unsolved problem. Compensation mechanisms may mask clear correlations between a lesion and the instability pattern associated with it. The proximal carpal row should be studied as one system. PMID- 8228051 TI - Evaluation of chronic wrist pain by arthrography, arthroscopy, and arthrotomy. AB - Arthrography and arthroscopy of the wrist were compared as diagnostic tools in the evaluation of chronic wrist pain. Arthroscopy proved to be a more sensitive procedure than arthrography in 20 consecutive patients with confirmed wrist injuries. There was 86% correlation (the exception--one false negative arthroscopic procedure) between arthroscopy and arthrography in eight patients with triangular fibrocartilage tears. Compared with arthroscopy, there were five false negative and five false positive arthrograms when evaluating interosseous scapholunate and lunotriquetral ligament tears. Arthroscopy more readily identified the location and extent of triangular fibrocartilage and the significance of interosseous ligament tears. Wrist arthrotomy confirmed the findings of wrist arthrography and arthroscopy and demonstrated that arthrography is not as accurate in localizing specific wrist injuries. Furthermore, wrist arthroscopy is a more valuable technique in determining the location, size, and extent of ligament injuries within the wrist. PMID- 8228052 TI - Toe-to-hand transfer: an analysis of 14 failed cases. AB - Fourteen failed cases of 325 patients who had undergone the operation of toe-to hand transfer to reconstruct the thumb and/or the finger are reported. Five key factors were related to the survival of the transferred toe: two sets of arterial blood supply in the case of an arterial anomaly, the resection of diseased vascular segments, the observation of capillary blood refill for the judgment of circulatory crisis, and the prevention and treatment of a hypercoagulable state. PMID- 8228053 TI - Longitudinal growth after replantation of immature extremities on rats. AB - Replantation of right hindlimbs in 76 immature rats was performed to study the factors affecting longitudinal growth of replanted extremities. There was a significant difference in tibia length between the replanted limb and the contralateral normal limb. The difference was significantly influenced by the ischemic interval (2 vs 4 hours), but not by suture method (intact vessels, vascular anastomosis with 10-0 nylon or 11-0 nylon). The replantation of immature extremities should be promptly undertaken because of the vulnerability of the epiphysis to ischemic insult. PMID- 8228054 TI - Toe phalangeal grafts in congenital hand anomalies. AB - The results of 73 toe phalangeal grafts were reviewed in order to determine the effects of age and operative technique on subsequent growth of the transferred phalanx. Minimum length of follow-up care for inclusion in the study was 1 year (mean, 42 months). Physeal openness and growth of transferred phalanges were determined from standard x-ray films. Physeal openness rates were 94% for those operated on before 1 year of age, 71% for those 1-2 years of age, and 48% for those older than 2 years of age. Mean growth rates were 1.0 +/- .2 mm, 1.0 +/- 0.6 mm, and 0.5 +/- 0.5 mm per year, respectively, for these three age groups. Extraperiosteal dissection of the transferred phalanx, tendon and collateral ligament reattachment, and patient age under 12 months had a statistically beneficial effect on maintaining physeal openness. PMID- 8228055 TI - Acrosyndactyly with epidermoid inclusion cysts: evidence for the extrinsic theory. AB - A case of acrosyndactyly was treated with routine syndactyly release and full thickness skin grafts. Two epidermoid inclusion cysts were found within the syndactylyzed index and middle fingers. This association is discussed along with the mechanism that best explains this unusual occurrence. PMID- 8228056 TI - Patterns of microvascular response associated with reflex sympathetic dystrophy of the hand and wrist. AB - The diagnosis of reflex sympathetic dystrophy is based on clinical presentation and often the presence of a positive three-phase technetium bone scan result. This study compared vasomotor control patterns between reflex sympathetic dystrophy patients with positive bone scan results and those with negative results. Both groups showed similar vasomotor responses to isolated cold stress testing--warm, cold, and transitional patterns of response were identified in both groups. Thus, this study demonstrates that a positive technetium bone scan result does not correlate with vasomotor disturbances in patients with reflex sympathetic dystrophy of the hand. PMID- 8228057 TI - Segmental reflex sympathetic dystrophy: clinical and scintigraphic criteria. AB - Clinical and scintigraphic criteria are proposed for the diagnosis of segmental reflex sympathetic dystrophy. Eight patients met previously described clinical criteria for reflex sympathetic dystrophy with involvement limited to only a portion of the hand. The delayed phase of the three-phase radionuclide bone scan was found to be highly sensitive (100%) for this small group of patients. Consecutive bone scans (n = 127) performed during a 6-month period for a variety of upper extremity problems were reviewed, and a segmentally diffuse pattern of tracer uptake was found to be highly specific (98%) for segmental reflex sympathetic dystrophy. Recognition and documentation of a more localized form of reflex sympathetic dystrophy will allow earlier recognition and treatment, which is an important factor in a successful outcome for managing pain dysfunction disorders. PMID- 8228058 TI - Effect of sympathetic block demonstrated by triple-phase bone scan. AB - The triple-phase bone scan is frequently used in the diagnosis of reflex sympathetic dystrophy although the etiology of the abnormal findings is not well established. This study evaluated the effect of increased blood flow secondary to sympathetic block on the triple-phase bone scan. In 15 reflex sympathetic dystrophy patients, all of whom had significant relief with sympathetic block, baseline and postsympathetic block bone scans were analyzed. There was a significant linear relationship between change in counts from baseline to postsympathetic block on the blood pool image and bone uptake image. As the blood pool and bone uptake counts increased, the likelihood of typical reflex sympathetic dystrophy findings on visual inspection also increased. These results strongly support the concept of increased vascular flow as the cause of increased periarticular uptake. PMID- 8228059 TI - Acute forearm compartment syndrome associated with HIV-induced thrombocytopenia. PMID- 8228060 TI - Abscesses of the upper extremity from drug abuse by injection. AB - A 4-year retrospective review of 59 consecutive upper extremity abscesses associated with drug abuse by injection is reported. There were 57 patients, with the most common location being the forearm. All abscesses were treated with incision, drainage, and intravenous antibiotics. Seventeen patients required more than one debridement; nine were complicated by fasciitis, osteomyelitis, septic arthritis, or septic tenosynovitis. Thirty-one patients had human immunodeficiency virus testing, and nine results were positive. Bacteriology showed the most common organisms to be streptococcus, Staphylococcus aureus, and Eikenella corrodens. Nineteen percent of the abscesses had anaerobes cultured. Most of the organisms cultured were common oral or skin flora. PMID- 8228061 TI - Upper limb involvement in patients with Gaucher's disease. AB - Seventeen of 55 children and adolescents with Gaucher's disease presented with upper limb pain. Seven had nonspecific pain, 11 had bone crisis, and 2 had arthritis symptoms in the proximal interphalangeal joints. X-ray films of both hands showed osteopenia and thinning of the cortex in half of the patients. PMID- 8228062 TI - Wrist and hand injuries in platform diving. AB - To permit a smooth entry into the water, platform divers assume a special position with their hands. An older method consists of hitting the water with both fists closed while holding one thumb in the palm of the other hand. In a newer technique fingers and wrists are hyperextended, the forearms pronated, and the hands overlapped. In this series of 21 divers, the injuries included 2 fractures and 5 sprains treated by conservative methods. Two divers required surgery: one with loose bodies in the radiocarpal articulation, the other with chronic rupture of the trapeziometacarpal ligament and synovitis of both extensor pollicis brevis and longus. Eighteen divers complained of pain caused by the impact of the hand with the water. A kinetic analysis with a video imaging system explains the mechanism and forces of water penetration. PMID- 8228063 TI - Triggering of the finger secondary to partial flexor tendon tear after closed direct injury. PMID- 8228064 TI - Use of botulinum toxin in the treatment of hand dystonia. AB - Forty-six patients with hand dystonia, considered disabling despite optimal pharmacologic therapy, were injected in the forearm musculature with botulinum A toxin. Thirty of these patients were followed long enough to provide adequate data for analysis of 86 treatment sessions. There was a 63% female preponderance, with an average age at initial evaluation of 46 years and symptom duration of 7.9 years. Average baseline severity of dystonia was rated as 3.5 on a severity rating scale (0-4 rating; 4 = maximum severity). The average peak effect response for all injections (79 into wrist flexors and 29 into wrist extensors) was 2.2 for dystonia and 3.0 for pain (0-4 rating; 0 = no response, 4 = maximum benefit). The latency from injection to onset of effect averaged 5.6 days. Total response duration averaged 9.3 weeks and maximum improvement was 7.5 weeks. Only local complications occurred and consisted primarily of hand weakness (25 patients, 44 sessions). The results show that botulinum toxin injections effectively control hand dystonia in instances where other forms of therapy have failed. PMID- 8228065 TI - Ulnar artery palmar to palmaris brevis: cadaveric study and three case reports. AB - Three cases in which the ulnar artery was palmar to the palmaris brevis muscle at the level of the wrist are described and the results of a cadaveric study are presented. The anomaly was confined to the level of Guyon's canal with a normal position of the artery proximally in the forearm and distally in the palm. In spite of its superficial location, the artery was patent in all patients and no symptoms referable to the ulnar artery were present. The ulnar nerve was found dorsal to the palmaris brevis in Guyon's canal in all three patients. An accessory muscle was found in one patient, but no other anatomic anomalies were seen. A cadaveric study of 43 limbs revealed a similar anomaly of the ulnar artery in 1 cadaver. In the presence of this anomaly the ulnar artery may be at increased risk during surgical exploration of Guyon's canal. Exposure of the artery in the forearm prior to exploration of Guyon's canal is recommended. PMID- 8228066 TI - Ulnar nerve compression at the arch of origin of the adductor pollicis muscle. AB - The motor branch of the ulnar nerve can be compressed by the arch of origin of the adductor pollicis muscle. There can be a well-defined band of tissue either at the point where the nerve crosses the third metacarpal or where it penetrates the adductor muscle. In over half of the cadaver hands studied no arch was demonstrated. Clinically these patients present with obvious atrophy of the first dorsal interosseous and adductor muscle. Usually pain is not a prominent symptom. This type of ulnar nerve compression neuropathy is present in less than 1% of those ulnar neuropathies that occur at the wrist and hand. PMID- 8228067 TI - Ulnar nerve transection as a complication of two-portal endoscopic carpal tunnel release: a case report. PMID- 8228068 TI - Treatment of chronic post-traumatic dorsal subluxation of the distal ulna by hemiresection-interposition arthroplasty. AB - Twenty-four patients were evaluated for complaints of pain in the area of the distal radioulnar joint, decreased forearm rotation, and weakness of grip. All patients had a history of trauma to the involved wrist an average of 40 months prior to presentation. Six patients had undergone previous surgery that was unsuccessful. Twenty-three patients were available for follow-up evaluation at an average of 36 months following hemiresection-interposition arthroplasty. At that time, results were excellent in 13 patients, good in 4 patients, fair in 3, and poor in 3. Two patients developed symptoms of styloidcarpal abutment (poor results), one of whom was upgraded to an excellent result after undergoing a shortening osteotomy of the ulna. PMID- 8228069 TI - Precision oblique osteotomy for shortening of the ulna. AB - The obstacles of prolonged healing time and technically demanding osteotomy and plate fixation in the performance of ulnar shortening osteotomies have been overcome by a precision system that includes a 45 degrees osteotomy and 2.7 mm interfragmentary lag screw. In 23 transverse osteotomies healing time averaged 21 weeks with one nonunion. In 17 precision oblique osteotomies healing time averaged a substantially shorter 11 weeks. Biomechanical data obtained from cadaveric testing comparing these two constructs demonstrated a structural stiffness that was clearly greater in torsion testing for the oblique osteotomy. No biomechanical difference was identified in the anteroposterior and lateral bending tests. The system permits the reliable performance of two parallel osteotomy cuts, allowing the removal of a precise amount of bone. The compression device and specialized plate permit easy coaptation of the osteotomy surfaces, which are locked into position by a precise 22 degrees interfragmentary lag screw. The surgical procedure is more quickly completed, and the frustration of this previously challenging procedure is now completely removed. PMID- 8228070 TI - Strains in the articular disk of the triangular fibrocartilage complex: a biomechanical study. AB - The articular disk of the triangular fibrocartilage complex was studied using a video imaging system in a cadaveric laboratory experiment. Changes in disk configuration consistently occurred during pronation and supination and resulted in a nonuniform strain distribution that was dependent upon forearm position. Strains occurred primarily in the radioulnar axis of the disk, with dorsoanterior strains being negligible. Strains were concentrated in the radial portion of the disk and were highest with the forearm pronated. Application of a distraction load to the distal radioulnar joint to simulate the effect of axial wrist loading caused strains to increase the most in the radial portion. These findings suggest that joint distraction loading and forearm pronation are important components of the injury mechanism for the most common type of traumatic tear that occurs near the radial attachment of the disk. PMID- 8228071 TI - Power staple fixation in trapeziometacarpal arthrodesis. AB - Twenty-seven trapeziometacarpal arthrodeses were performed in 26 patients for basilar thumb joint arthritis utilizing power staple fixation. Short arm thumb spica cast immobilization was maintained for an average of 8 weeks, followed by part-time splintage for 6-12 weeks. Twenty patients were available for evaluation an average of 19 months after the operation. All of the patients were satisfied with the procedure. Thirteen had complete relief of pain, four had mild pain, and three had moderate pain after surgery. The two nonunions that occurred were in patients that refused postoperative casting and both had complete pain relief. There were no fixation-related complications in the remaining 18 patients that were casted postoperatively, and all developed a solid fusion. Power staple fixation in trapeziometacarpal arthrodesis has a 90% union rate with no long-term fixation related complications. PMID- 8228072 TI - Ligament replacement for chronic instability of the ulnar collateral ligament of the metacarpophalangeal joint of the thumb. AB - Static and dynamic procedures have been described for reconstruction of chronic instability of the ulnar collateral ligament of the thumb metacarpophalangeal joint. This study presents a technique of ligament replacement utilizing a free tendon graft passed through two gouge tracks in the proximal phalanx and one in the metacarpal in a manner that closely approximates normal anatomy. We retrospectively reviewed 26 patients who underwent replacement. The follow-up period averaged 4.5 years. In 24 of 26 cases the joint was rendered stable by the replacement and the patient was relieved of pain. Eighty-five percent of the arc of motion was maintained. Postoperative key pinch measured 20 lb. on the operated side compared to 21 lb. on the unoperated side. Results were excellent in 20 patients, good in 4, and fair in 2. This technique successfully restores stability to the ulnar collateral ligament of the thumb metacarpophalangeal joint, diminishes pain and weakness with minimal loss of motion, and holds up over time. PMID- 8228073 TI - Technique for bone approximation in ulna shortening. PMID- 8228074 TI - Simultaneous trapezium and Bennett's fractures. PMID- 8228075 TI - Molecular gastroenterology--implications for medical practice. AB - Molecular biology has revolutionized our understanding of the life of the cell. The development of concepts has been accompanied by the development of new technologies, many of which have clinical applicability. This new armamentarium includes methods for detection and amplification of minuscule amounts of DNA, automated methods for DNA and RNA sequencing, the ability to express a gene in cultured cells and harvest its product, approaches to identify genes whose product is unknown, methods for a detailed understanding of the regulation of gene expression, and innovative methods for gene therapy. These developments have already had an impact on gastroenterology, exemplified by the identification of hepatitis viruses and the agent causing Whipple's disease, development of a recombinant vaccine for hepatitis B, and isolation of the cystic fibrosis gene, which will lead to the diagnosis and treatment of this disease. This editorial outlines these recent changes in the conceptual and practical framework of gastroenterology. PMID- 8228076 TI - NSAID-induced ulcers and prophylaxis. A reappraisal. AB - The most common side effect of nonsteroidal anti-inflammatory drug (NSAID) therapy involves gastrointestinal damage that may involve hemorrhages and petechiae to erosions and ulcers. The NSAID usage increases the risk for ulcer complications such as bleeding and perforation. While prophylaxis against NSAID induced ulcers is an appealing clinical strategy, the studies to date concerning its effectiveness have been flawed by uneven experimental and statistical design. Important questions concerning management of NSAID-induced ulceration have yet to be answered. PMID- 8228077 TI - Cimetidine and ranitidine. Lack of effect on the pharmacokinetics of an acute ethanol dose. AB - The effect of H2-receptor antagonists on alcohol absorption appears to vary, depending on testing conditions and subject population. In this crossover study of cimetidine (400 mg b.i.d.) and ranitidine (150 mg b.i.d.), we evaluate alcohol pharmacokinetics in 10 subjects after ingestion of moderate doses (0.5 g/kg) of ethanol under four challenge conditions: (a) a single dose of placebo prior to alcohol ingestion in the morning, (b) a single dose of H2-antagonist prior to alcohol ingestion in the morning, (c) 14-day chronic dosing with H2-antagonist prior to alcohol ingestion in the evening, and (d) 28-day chronic dosing with H2 antagonist prior to alcohol ingestion in the morning. No significant increases were observed for alcohol in the area under the curve (AUC) or Cmax. Significant decreases in AUC (16 and 13%) were observed following administration of single- and multiple-dose cimetidine, and a significant decrease in Cmax (14%) was observed following a single dose of ranitidine--all given prior to alcohol ingestion in the morning. These results do not support any clinically meaningful effect of H2-antagonists on alcohol absorption following ingestion of a moderate dose of ethanol. PMID- 8228078 TI - Endoscopic injection therapy of bleeding ulcer: a prospective and randomized comparison of adrenaline alone or with polidocanol. AB - In a prospective randomized trial involving 63 patients with bleeding peptic ulcer, we assessed whether the addition of 1% polidocanol improved the results achieved by 1/10(4) adrenaline alone for injection therapy. The inclusion criterion was the presence of active arterial bleeding or a nonbleeding visible vessel at emergency endoscopy. Thirty patients were treated with 1/10(4) adrenaline (group A) and 33 with adrenaline plus 1% polidocanol (group B). Initial hemostasis was achieved in 97% of cases in both groups and permanent hemostasis in 87% patients in group A and in 76% in group B (p = NS). Mortality was 6% in group A and 3% in group B. There were no differences between the two groups regarding requirements for emergency surgery, the number of transfusions, or the length of hospital stay. One patient in group B had a perforation. No other relevant complications were noted. In conclusion, combined therapy does not improve the results achieved with adrenaline alone. PMID- 8228079 TI - Mucosal blood flow in erosive duodenitis. AB - Erosive duodenitis is a clinical entity whose pathogenesis is still obscure. Acid secretion is mostly normal, while the possible role of vascular factors has been suggested. We measured mucosal blood flow by means of laser Doppler velocimetry during endoscopy in the duodenal bulb of 10 subjects with erosive duodenitis, 10 duodenal ulcer patients, and 10 healthy controls. Duodenal blood flow in erosive duodenitis was significantly reduced (p < 0.001) compared with controls, whereas no changes were detected in duodenal ulcer. Reassessment of mucosal blood flow after a 6-week treatment with ranitidine failed to show any increase in the perfusion values even in patients with endoscopic healing. Our results suggest that impaired mucosal blood flow is a primary factor in the pathogenesis of chronic duodenal erosions. PMID- 8228080 TI - Is gastric metaplasia essential for duodenal ulcer? AB - Multiple biopsies were taken from the duodenum of 100 patients (46 active duodenal ulcer, 14 healed duodenal ulcer, 40 control subjects) to detect duodenitis, gastric metaplasia and Helicobacter pylori (Hp). Histamine-induced congo-red staining of the duodenal bulb was performed in additional 25 cases of active duodenal ulcer to determine the presence of functioning parietal cells in the duodenum. The incidence of duodenitis was 45% in control subjects and 76.7% in patients with duodenal ulcer (active or healed) (p < 0.01). Superficial gastric metaplasia was present in 2.5% of control subjects and 13.3% patients with duodenal ulcer (active or healed) (p > 0.05). Hp in the duodenum was detected in 10% of control subjects and 8.3% of patients with duodenal ulcer. Congo-red test was positive in two of 25 (8%) patients of active duodenal ulcer indicating "functioning" parietal cells in the duodenum. In Indian patients with duodenal ulcer, a low incidence of gastric metaplasia and Hp in the duodenum is observed in contrast to its high incidence in patients from Western countries. PMID- 8228081 TI - Proctitis and rectal stenosis induced by nonsteroidal antiinflammatory suppositories. AB - Anorectal ulceration eventually leading to rectal stenosis was observed in 10 patients who abused analgetic suppositories containing acetylsalicylic acid, acetaminophen, and codeine. Most patients were middle-aged women with a neurotic or psychiatric background. Perianal skin lesions were present in half of the patients. The endoscopic aspect of the sharply demarcated distal rectal lesions with squamous and transitional epithelium (anoderm) ascending from the anal canal was rather typical, but the appearances on biopsy were nonspecific. Possible pathophysiological mechanisms include mucosal prostaglandin depletion secondary to blockade of cyclooxygenase and shift to lipoxygenase products in the arachidonic acid cascade. Treatment options include drug cessation, topical steroids, and endoscopic dilation. Supporting psychological therapy often is necessary. PMID- 8228082 TI - Experience with Eudragit-S-coated mesalamine (Asacol) in inflammatory bowel disease. An open study. AB - We reviewed the courses of 63 patients with inflammatory bowel disease (IBD) treated in a private gastroenterology practice. All were allergic, unresponsive, or in some way intolerant of sulfasalazine and therefore were treated with Eudragit-S-coated mesalamine (Asacol). Thirty-eight patients had ulcerative colitis (UC) and 25 had Crohn's disease. In 22 and 14, respectively, mesalamine was used to treat active disease; in 16 and 11, respectively, it was introduced to maintain remission. In some patients, mesalamine was used along with other medications. Symptomatic and endoscopic response was seen in seven of nine (77%) with active UC treated with measalamine alone; remission was maintained in seven of eight (94%) when mesalamine was used alone. Therapeutic success was noted in eight of 10 (80%) with active Crohn's disease, and remission was maintained in seven of nine (78%) when mesalamine was used alone. Adverse reactions of varying severity occurred in 21 of 63 (33%), but the drug had to be terminated in only 9 of 63 (14%). Eudragit-S-coated mesalamine appears to be effective in the treatment and maintenance of remission of UC in 82% of those patients allergic or intolerant to the parent drug, sulfasalazine. Furthermore, contrary to what was expected, we found it effective in a much larger percentage of patients with Crohn's disease (79%) than the parent drug. PMID- 8228083 TI - Clinical implications of liver biopsy findings in collagen-vascular disorders. AB - We surveyed over a 5-year period liver abnormalities in all patients with collagen-vascular disorders in whom liver histology was obtained, including 46 patients with systemic lupus erythematosus, rheumatoid arthritis, primary Sjogren's syndrome, periarteritis nodosa, mixed cryoglobulinemia, Wegener's granulomatosis, systemic sclerosis, and other conditions. Histological appearances diagnostic of the primary condition were only found in three patients, each of whom had periarteritis nodosa. Significant chronic liver disease was found in 11 patients (24%), in five of whom a strong clinical suspicion of severe chronic liver disease already existed. Clinically inapparent but potentially significant chronic liver disease was found predominantly in patients with mixed cryoglobulinemia or sicca syndrome. Seventeen percent of all biopsies suggested drug induced hepatitis. In patients with collagen-vascular diseases and abnormal liver function tests, histological examination of the liver is most frequently of value in indicating drug-induced liver damage. Significant chronic liver disease is common but usually clinically apparent. In patients with periarteritis nodosa and mixed cryoglobulinemia, liver biopsy may be of value diagnostically, revealing serious liver disease with prognostic and therapeutic implications. PMID- 8228084 TI - Increased plasma endothelin-1 levels in patients with cirrhosis and esophageal varices. AB - We measured plasma levels of endothelin-1, a potent vasoconstrictor peptide, in 6 healthy controls and 20 patients with cirrhosis and correlated them with various clinical and laboratory parameters and other vasoactive substances. There was a trend toward increased endothelin-1 levels in patients with cirrhosis compared with controls (6.0 +/- 2.5 vs. 4.4 +/- 1.5 pg/ml, NS). The presence of ascites did not influence plasma endothelin-1 levels, and plasma endothelin-1 levels were not significantly correlated with serum albumin, serum bilirubin, or prothrombin time. Plasma endothelin-1 levels were significantly elevated in cirrhotic patients with esophageal varices compared with those without varices (7.5 +/- 2.5 vs. 4.5 +/- 1.6, p < 0.05). The elevation of endothelin-1 levels in patients with varices may represent a physiological compensation for the systemic vasodilation present in patients with liver cirrhosis and portal hypertension. PMID- 8228085 TI - Spontaneous reactivation of chronic hepatitis B infection leading to fulminant hepatic failure. Report of two cases and review of the literature. AB - Reactivation of chronic hepatitis B viral (HBV) infection, defined as conversion from the low level replicative phase (HBV-DNA negative) to the high level replicative phase (HBV-DNA positive) with continued hepatitis B surface antigen positivity, occurs after immunosuppressive therapy, chemotherapy, and rarely spontaneously. Development of hepatic failure after spontaneous reactivation of chronic HBV is rare. We report two patients with chronic HBV infection and stable cirrhosis in whom spontaneous reactivation lead to hepatic failure. In one, spontaneous reactivation lead to severe jaundice, ascites, and encephalopathy. Treatment with alpha-interferon resulted in transient improvement, but the patient ultimately died with hepatorenal syndrome. In the second, severe liver failure required orthotopic liver transplantation. Spontaneous reactivation is an unusual cause of hepatic decompensation in patients with chronic HBV infection and may lead to fulminant hepatic failure. The role of interferon or liver transplantation in the management of spontaneous reactivation remains to be clarified. PMID- 8228086 TI - Is there a role for antibiotics as primary therapy in Crohn's ileitis? AB - Although their use has been deemed plausible by previous investigators, the role of antibiotics as primary therapy in active Crohn's ileitis is still unclear. Here we detail the response of four patients with active ileitis to ciprofloxacin, a quinolone antibiotic. Each patient had a dramatic improvement in abdominal pain and diarrhea coincident with the institution of ciprofloxacin, despite the absence of any obvious enteric infection. It is uncertain whether ciprofloxacin is treating an undetected pathogen, bacterial overgrowth, an unsuspected microperforation, or even if it is having its effect through a mechanism entirely unrelated to its antimicrobial properties. Our experience suggests that there is a need for a prospective controlled trial of ciprofloxacin or other antibiotics in active Crohn's ileitis. PMID- 8228087 TI - MRI in the diagnosis of Crohn's disease in two pregnant women. AB - Two pregnant women considered symptomatic for inflammatory bowel disease were referred for magnetic resonance imaging (MRI) for assistance in establishing a diagnosis. Breath-hold gradient echo and contrast-enhanced T1-weighted fat suppressed spin echo sequences were used. The MR appearance of the terminal ileum was consistent with Crohn's disease in both patients. The diagnosis of Crohn's disease was subsequently confirmed histologically. MRI is a safe, noninvasive modality that may be useful in confirming a suspected diagnosis of inflammatory bowel disease, especially in patients who cannot be evaluated with standard techniques. PMID- 8228088 TI - Magnetic resonance imaging evaluation of the local extent of colorectal mass lesions. AB - Magnetic resonance imaging (MRI) was conducted in 21 patients with known or suspected colorectal mass lesions. Imaging was carried out at 1.5 T using Fast Low Angle Shot (FLASH) and fat-suppressed (T1FS) gadolinium enhanced sequences. The lesions studied included adenocarcinoma (n = 16), lipoma (n = 2), villous adenoma with foci of adenocarcinoma (n = 1), malignant melanoma (n = 1), and rectal lymphoma (n = 1). Correlation was made with surgical findings in 14 patients, 10 of whom had tumors resected. Bowel wall involvement, tumor size, and extension demonstrated on MR images was correlated with histopathology findings. No significant differences were found (p > 0.05) among those parameters when compared to MR information from T1FS images. Lymph nodes were demonstrated on MR images but distinction between benign and malignant nodes could not be made. PMID- 8228089 TI - Pancreatic disease in AIDS--a review. AB - Patients with the acquired immunodeficiency syndrome (AIDS) can develop pancreatic disease from causes unrelated to AIDS as well as AIDS-specific lesions. AIDS-specific causes include opportunistic infection, AIDS-associated neoplasia, and medications used to treat complications of AIDS. Reported pancreatic opportunistic pathogens include Mycobacterium tuberculosis, Mycobacterium avium intracellulare, Cryptococcus neoformans, Candida, Aspergillus, Toxoplasma gondii, Pneumocystis carinii, cytomegalovirus, herpes simplex, cryptosporidium, and microsporidium. Although cytomegaloviral pancreatic infection can occur without clinically evident pancreatic disease, cytomegalovirus can cause pancreatitis. Other opportunistic infections that can cause pancreatitis include Toxoplasma gondii, Cryptococcus neoformans, and Candida. Mycobacterial infection can produce a pancreatic abscess. Hepatobiliary or pancreatic duct infection by cytomegalovirus, cryptosporidium, and microsporidium causes irregular ductular narrowing and dilatation. This cholangiographic abnormality resembles the pattern found in idiopathic sclerosing cholangitis. Reported AIDS-associated pancreatic neoplasms include Kaposi's sarcoma and lymphoma. Pancreatic involvement is usually part of widely disseminated tumor and rarely produces clinical symptoms. Pentamidine, trimethoprim-sulfamethoxazole, and 2', 3'dideoxyinosine are medications commonly used in AIDS patients which can cause pancreatitis. Pentamidine also causes hypoglycemia or hyperglycemia. PMID- 8228090 TI - A case of squamous papilloma of the esophagus: spontaneous disappearance? PMID- 8228091 TI - The problem of chronic nonspecific gastritis: classification today. PMID- 8228092 TI - Giant malignant cystic leiomyoblastoma of the stomach imitating hemorrhagic ascites. PMID- 8228093 TI - Carcinoma of the sigmoid colon associated with Sjogren's syndrome. PMID- 8228094 TI - H. pylori in postcholecystectomy symptoms. PMID- 8228095 TI - Salmonellosis-associated pancreatitis and pancreas divisum? PMID- 8228096 TI - Hydrothorax complicating cirrhosis in the absence of ascites. PMID- 8228097 TI - Neurobiology of bowel habits in Israeli women. PMID- 8228098 TI - Airway management, penetrating neck trauma. AB - This study was performed to determine whether clinical presentation can accurately predict which victims of penetrating neck trauma require urgent airway management. An 8-year retrospective review of all patients with a diagnosis of penetrating neck trauma seen in the emergency department of an urban teaching hospital was conducted. Of the 114 patients reviewed, 69 (60%) were intubated at some point in their hospital course. Twenty-six (23%) met our predetermined criteria for urgent airway control; 25 of these patients were intubated in the emergency department. Forty-three patients (38%) did not meet the criteria and were electively intubated either in the operating room or in the emergency department. Forty-five patients (39%) were never intubated. None of the patients in this series developed complications as a result of their airway management. PMID- 8228099 TI - Fiberoptic intubation in the emergency department: a case series. AB - A prospective descriptive study was conducted to evaluate the feasibility of using fiberoptic bronchoscopy to replace blind nasotracheal intubation. The study was performed from November 1989 to August 1991 at a university teaching tertiary care hospital. Intubations were performed by three senior emergency physicians with no prior clinical experience in emergency fiberoptic intubation. Training in fiberoptic intubation was carried out on anesthetized patients, intubation manikins, and cadavers. All patients coming to the emergency department who would have been intubated in the blind nasotracheal manner were eligible for fiberoptic intubation, except for apneic patients and those intubated by residents learning other techniques. An independent observer collected the data. Forty-two patients were entered into the study, 22 males and 20 females, with an average age of 64 years. The success rate was 72% (30/42), with one of the investigators performing most of the intubations (22, 52%). His success rate was 82%. The other two investigators' success rates were 64% and 56%. Most failures were ascribed to coping with secretions. In conclusion, intubation with the fiberoptic bronchoscope can be a useful alternative to blind nasotracheal intubation. Success with this technique is dependent on the airway being free of secretions and blood. PMID- 8228100 TI - Reflex sympathetic dystrophy after a minor electric shock. AB - A 37-year-old right-handed female sustained a minor electric shock to her right hand while changing a bulb in a Kodak Reliant Intelligent Microfilmer 2000. She noted a progressive onset of parasthesias of her right hand that became associated with throbbing pain at rest, as well as swelling and profound weakness of her hand during the next two months. Plethysmography demonstrated arterial vasoconstriction of her right hand. Her reflex sympathetic dystrophy was successfully treated with cervicothoracic sympathetic blocks. PMID- 8228101 TI - Immersion scald burns: strategies for prevention. AB - Tap water scald burns are common injuries to persons with disabilities, young children, and the elderly. A case is reported of an elderly woman with a physical and neurological handicap who while bathing received partial and full thickness (tap water) scald burns covering 20% of her total body surface area. This life threatening injury could have been prevented with a Shower Safe, Inc. temperature controlling water valve. PMID- 8228102 TI - Embolization of illicit needle fragments. AB - Complications encountered with retained or embolized foreign bodies (FB) are mainly reported in association with invasive intravascular monitoring or diagnostic devices. Retrieval of these large FB is recommended. On the other hand, the embolization of hypodermic needle fragments in the intravenous drug abuser is infrequently reported. The natural history and management of this complication is controversial. To date, only ten cases have been reported in the literature. We present a case of a hypodermic needle fragment that embolized to the lung from a forearm vein of an intravenous drug abuser. The embolization occurred during an attempt at removal of the needle fragment from the vein at the injection site. The needle fragment lodged in the lung parenchyma and was left in situ. No complications related to the needle occurred during a 10-month follow up. PMID- 8228103 TI - Unrecognized pathologic fractures of the cervical spine: two case reports and review of the literature. AB - Two cases of undiagnosed pathologic fractures of the cervical spine are presented. A discussion of the pathogenesis, typical presentation, and the optimal emergency department evaluation of a possible pathologic fracture of the cervical spine follows the reports. PMID- 8228104 TI - Theophylline toxicity. AB - Theophylline toxicity continues to be a commonly encountered clinical problem. Patients may present with a vast array of toxic manifestations, including life threatening cardiovascular and neurologic toxicity. Despite the considerable attention this topic has received in the literature, there remain some important controversies regarding the identification of high risk patients and how best to manage them. This review attempts to summarize the current state of knowledge regarding theophylline toxicity with special emphasis on toxic manifestations and the role of elimination enhancing modalities. PMID- 8228105 TI - Theophylline toxicity from an over-the-counter preparation. AB - The clinical presentation of theophylline poisoned patients has been well described in the literature. These individuals may develop severe and potentially fatal cardiac, neurologic, and gastrointestinal manifestations. While patients may present following an intentional over-dose, a significant percentage become toxic accidentally or iatrogenically, as a result of theophylline's narrow therapeutic index. Another factor, not well known or described in the literature, is the availability of theophylline in a variety of over-the-counter formulations. We present a case of theophylline toxicity from a nonprescription combination product containing theophylline, ephedrine, and phenobarbital. Clinicians should be aware of the potential for serious toxicity from over-the counter medications, particularly those commonly thought of as "prescription only." PMID- 8228106 TI - Prehospital misidentification of tachydysrhythmias: a report of five cases. AB - Identification and treatment of tachydysrhythmias is an important element of prehospital care. Five cases of prehospital misidentification of tachydysrhythmias are presented to highlight the challenges and pitfalls of field management. The literature is reviewed and discussed in light of new therapies and technology. Guidelines for patient subgroups potentially benefitting from on line medical control and biotelemetry are explored. PMID- 8228107 TI - Acute spinal epidural hematoma. AB - A 17-year-old female presented to the Emergency Department with a history of recent onset of severe upper thoracic back pain, urinary retention, weakness and paraesthesia in the lower limbs. Magnetic Resonance Imaging (MRI) Scan demonstrated a spinal epidural hematoma at the C7-T2 level. This report highlights a rare, but eminently treatable condition; the use of MRI Scan as a specific diagnostic tool; and the need for urgent surgical decompression to ensure good recovery of function. PMID- 8228108 TI - The roentgenographic findings associated with air embolism in sport scuba divers. AB - Records on all patients with arterial gas embolism (AGE) presenting to UCSD from 1982-1989 and for whom chest radiographs were available were reviewed. Of the 31 patients, 13 roentgenograms (42%) showed evidence of pulmonary barotrauma demonstrated by pneumomediastinum (N = 8), subcutaneous emphysema (N = 3), pneumocardium (N = 2), pneumoperitoneum (N = 1), or pneumothorax (N = 1). Pneumopericardium was not seen. Sixteen (52%) of the 31 patients had pulmonary infiltrates. Radiographic evidence of barotrauma was on occasion subtle, and in four cases was overlooked. Evidence of barotrauma (i.e., extra-alveolar air) was often identified along the left cardiac border, aortic arch, descending aorta, and hilar vessels. Subtle findings of ectopic air can confirm the clinical diagnosis of AGE; however, radiographic evidence of concomitant near drowning occurs more frequently. PMID- 8228109 TI - Technical considerations in the selection of a personal emergency response system. AB - It is the purpose of this report to provide an internal review of the personal response system (PRS) in our hospital. Our system is coordinated by volunteers without emergency physician supervision. It uses antiquated equipment in the client's homes and emergency response center. Calls are answered by individuals who have not been trained as dispatchers, and who do not record pertinent performance data. The technical considerations involved in the selection and operation of an ideal PRS system are described. PMID- 8228110 TI - Electric shock, Part II: Nature and mechanisms of injury. AB - Virtually every part of the body can be injured by electric current. The extent of injury to any given tissue will depend on many factors, including the nature of the tissue and the amount and duration of the electric current. In addition, cardiac and respiratory arrest can be induced by a number of mechanisms with little or no immediate tissue damage. Burns can be caused by the heating of tissue by electric current and by other mechanisms. Secondary trauma may result from falls, explosions, and other events initiated by electric forces. Nervous tissue has the least resistance to current flow and is thus more easily damaged. Part II in this 3-part series will discuss nervous tissue damage first, followed by discussions of damage to tissues of increasingly greater electric resistance. These are blood vessels, muscle, skin, and bone. Less common injuries are discussed last. PMID- 8228111 TI - Alcohol-related seizures, Part I: Pathophysiology, differential diagnosis, and evaluation. AB - Alcoholism may be society's most devastating problem short of war and malnutrition. Perhaps the most complex and perplexing medical complication of alcoholism is alcohol-related seizures. This article is a collective review designed to provide emergency physicians with an overview of the topic that is pertinent to their clinical practice. Part 1 addresses the pathophysiology, differential diagnosis, and evaluation of alcohol-related seizures. Part 2 will concentrate on the clinical presentation, management, and disposition. In addition, a classification of alcohol-related seizures will be proposed. PMID- 8228112 TI - Fools rush in? Airway management in penetrating neck trauma. PMID- 8228113 TI - Theophylline toxicity. PMID- 8228114 TI - Examining the agreement between triage nurses and emergency physicians in ordering extremity X-rays in children. PMID- 8228115 TI - Objectives to direct the training of emergency medicine residents on off-service rotations: hyperbaric medicine. AB - Objectives for Emergency Medicine residents on Hyperbaric Medicine rotations is the 22nd article in this continuing series of "off-service" goals and objectives. The role of the emergency medicine physician in hyperbaric treatment is evolving as part of a multidisciplinary approach to patients with widely divergent disease states who may benefit from serum hyperoxia and compression. Therefore, emergency residents with interests in this area will benefit from clear goals and objectives during a 2 to 4-week Hyperbaric Medicine Elective. PMID- 8228116 TI - Hocus-pocus: the art of medicine. PMID- 8228117 TI - Bacterial infections in patients with cirrhosis. PMID- 8228118 TI - Autoimmune hepatitis: the lesson of the discovery of hepatitis C virus. PMID- 8228119 TI - Colon cancer seeding after percutaneous fine needle aspiration of liver metastasis. AB - Fine needle aspiration or biopsy is used for cytologic diagnosis of many intra abdominal tumors including focal liver lesions. The incidence of needle tract seeding is quite low. In this paper the first case of cutaneous seeding after percutaneous fine needle aspiration of liver metastasis is reported in a case of colon cancer. We suggest using this method for cytologic diagnosis in hepatic tumors when surgical resection is not possible and when patients will be treated with invasive therapies and to avoid fine needle aspiration biopsy in patients undergoing surgical resection or when there is a confident diagnosis of HCC by non-invasive procedures. PMID- 8228120 TI - Frequency and severity of HCV infection following orthotopic liver transplantation. Effect of donor and recipient serology for HCV using a second generation ELISA test. AB - HCV is the principal etiologic agent of post-transfusion (PTH) hepatitis. The incidence and course of HCV hepatitis in liver transplant recipients is not well established. To resolve this information deficit, all records of recipients of single liver transplant (OLTx) between March 1986 and March 1990 at the University of Pittsburgh in whom both the donor and recipients' pre-OLTx sera were available (n = 516) were reviewed. All sera were assayed for HCV antibody using a second generation ELISA method developed by Abbott Laboratories. On the basis of the anti-HCV status of the donor and recipient pre-OLTx sera, four groups could be classified: group I (donor-, recipient-) n = 375; group II (donor , recipient+) n = 111; group III (donor+, recipient-) n = 25; and group IV (donor+, recipient+) n = 5. Post OLTx liver biopsies were obtained for a clinical indication in 473 of these 516 patients. The prevalence of anti-HCV among recipients pre-OLTx was 22.5% (116/516) which is three times greater than the 5.8% (30/516) prevalence in the donors. Histologic hepatitis not ascribable to any cause other than HCV occurred in 76/516 (15%) recipients: 42 in group I; 28 in group II; 6 in group III and none in group IV. The overall risk of HCV hepatitis at 6 months, 1 year and 2 years post-OLTx was 4.8% (25/516), 7.6% (39/516) and 10.1% (52/516), respectively. At each of these time intervals, no significant difference between groups for the prevalence of HCV hepatitis was evident.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228121 TI - Listeria monocytogenes hepatitis in a liver transplant recipient: a case report and review of the literature. AB - Listeria is an uncommon cause of hepatitis in adults. We report the case of a liver transplant recipient who presented with a clinical picture of acute hepatitis, 8 months after grafting. Blood cultures yielded Listeria monocytogenes. The patient made a full clinical recovery after adequate antimicrobial therapy (ampicillin and gentamicin intravenously for 4 weeks). Hepatitis was attributed to the Listeria infection. We believe this is the first reported case of Listeria hepatitis in an organ transplant recipient. PMID- 8228122 TI - Infectious sequelae after endoscopic sclerotherapy of oesophageal varices: role of antibiotic prophylaxis. AB - In order to determine the incidence of infection following sclerotherapy and the role of antimicrobial prophylaxis, a prospective randomized control study was performed comparing i.v. imipenem/cilastatin, with an infusion of dextrose-saline as a control group. One hundred patients with bleeding esophageal varices were included. All episodes of infection were documented during admission to the unit. Ninety-seven patients were evaluable. Post-sclerotherapy bacteremia developed in six (5.6%) of 107 sclerotherapy sessions in the control group and one (1.1%) of the 88 sclerotherapy sessions in the imipenem/cilastatin group (P < or = 0.1, NS): six of these seven post-sclerotherapy bacteremias occurred after emergency sclerotherapy. Infection within 7 days of the procedure was documented after 43 (22.1%) of the 195 sclerotherapy sessions, 18 (20.5%) in the imipenem/cilastatin group and 25 (23.4%) in the control group (P = NS). These infections were significantly more common after emergency sclerotherapy, 40 (34.8%) of 115 sessions, than after elective sclerotherapy, three (3.8%) of 80 sessions (P < or = 0.0001). A short prophylactic antibiotic regime does not reduce the risk of early bacteremia or the frequency of infection after sclerotherapy. The higher risk of infection after emergency sclerotherapy may be therefore related more to the gastrointestinal hemorrhage and its associated effects than to sclerotherapy. PMID- 8228123 TI - Increased frequency of CD8+ CD45R0+ memory T lymphocytes in acute hepatitis B virus infection. AB - CD8+ lymphocytes of the memory subset (= CD45R0+) encompass antigen-specific effector cells, which are believed to be decisive for virus elimination in several viral infections. To determine whether this can be extended to HBV infection naive and memory T cells were studied among CD(4+)- and CD(8+) lymphocytes and used monoclonal antibodies in two-color flow cytometric analysis to quantitate functional T cell subsets in peripheral blood of patients with acute hepatitis B (n = 11), chronic hepatitis B (n = 24) and healthy individuals (n = 26). Compared to CD4+ populations of healthy individuals the number of total CD4+ lymphocytes in patients with both acute or chronic hepatitis was significantly reduced. In contrast CD8+ cells did not significantly change in either acute and chronic hepatitis. Analysis of naive and memory subsets demonstrated, however, a significant rise in CD45R0+ memory cells from 5 to 15% (70% of all CD8+ cells) in acute hepatitis. These changes within the CD8+ population were, however, restricted to the acute phase of hepatitis in that the frequency of CD8+CD45R0+ decreased within weeks post infection. Furthermore, patients with chronic hepatitis did exhibit normal values of CD8+ memory cells (30% of all CD8+ cells). These findings suggest that enrichment of CD8+CD45R0+ memory cells reflects an accumulation of functional effector cells, which may be specifically activated by viral antigens and determine the outcome of infection. PMID- 8228124 TI - Regulation of ferritin and transferrin receptor expression by iron in human hepatocyte cultures. AB - HepG2 cell cultures and human hepatocyte primary cultures were used to develop appropriate hepatocytic in vitro models of iron load in order to further understand the pathophysiological mechanisms occurring in the liver of patients with hemochromatosis. The first step of this study was to obtain an efficient iron supply in conditions of minimal toxicity. It was demonstrated that iron complexed to citrate entered efficiently into HepG2 cells and human hepatocytes. This iron load was obtained with minimal toxicity in both culture models as evaluated by the intracellular LDH activity and the total protein content. The second step was to study the effect of iron on ferritin and transferrin receptor expression. In HepG2 cell cultures, intracellular and extracellular ferritin concentrations were strikingly increased by iron in dose- and time-dependent manners. However, the relative amounts of H and L ferritin mRNAs were not significantly affected by iron, suggesting that ferritin regulation occurred at a translational level. On the other hand, in human hepatocyte cultures, the increase of intracellular and extracellular ferritin concentrations was accompanied by an increase in the amounts of H and L ferritin mRNAs. In this model, iron-induced ferritin biosynthesis seemed to be more complex than in HepG2 cells and to be governed by transcriptional and/or post-transcriptional regulatory mechanisms. However, an additional translational level of regulation could not be excluded. In contrast, transferrin receptor expression was decreased by iron in HepG2 cells as well as in human hepatocyte cultures. This decrease was associated with a decrease in the mRNA steady-state level. In both culture models, transferrin receptor regulation seemed to occur at a transcriptional or post-transcriptional level. These results demonstrate that normal human hepatocytes in primary culture respond to iron in a manner close to that observed in vivo and thereby provide a promising experimental model for further understanding pathophysiological mechanisms involved in human hemochromatotic liver. PMID- 8228125 TI - The molecular basis of alpha 1-antichymotrypsin deficiency in a heterozygote with liver and lung disease. AB - Alpha 1-antichymotrypsin (alpha 1-ACT) is a serine proteinase inhibitor (serpin) with cathepsin G, mast cell chymase and chymotrypsin as target enzymes. We present the case of a middle-aged man with low plasma levels of alpha 1-ACT, asthma with progression to emphysema, and chronic HCV positive liver disease with selective accumulation of alpha 1-ACT in hepatocytes. This secretory defect is analogous to that seen in Pi Z alpha 1-antitrypsin deficiency. The molecular basis of alpha 1-ACT deficiency in this patient has been characterized by direct sequencing of the alpha 1-ACT genes from the patient and his father. A C-->G transversion in exon III causing a 229Pro-->Ala substitution is proposed to cause a conformational change resulting in abnormal transport through the RER. This mutation was found in one of 20 additional tested patients with chronic obstructive lung disease, but in no control. Two additional polymorphisms of the gene have been identified in unrelated healthy individuals with normal plasma alpha 1-ACT levels. The alpha 1-ACT deficiency state may predispose to obstructive lung disease and influence the course of liver disease. Identification of a specific mutation allows identification of heterozygotes for this deficiency allowing future evaluation of its clinical significance. PMID- 8228126 TI - Serum collagenase activity in patients with chronic liver disease. AB - To examine the clinical significance of serum collagenase activity in chronic liver disease, serum collagenase activity was determined in 50 patients with chronic liver disease and in 24 healthy controls. Collagenase activity was measured after reactivation by denaturing and dissociating the inhibitors with potassium thiocyanate and aminophenylmercuric acetate. In patients with chronic persistent hepatitis, serum collagenase activity was 37% lower than controls, 50% lower in those with chronic active hepatitis, 66% lower in those with cirrhosis and 68% lower in those with hepatocellular carcinoma. Serum collagenase activity was significantly and inversely correlated with serum levels of the aminoterminal propeptide of type III procollagen and type IV collagen 7S domain, indicating that serum collagenase activity decreased as liver active fibrogenesis and/or fibrosis occurred. In contrast, serum levels of the metalloproteinase inhibitor was 30% higher than controls in patients with chronic active hepatitis, 50% higher in those with cirrhosis and 80% higher in those with hepatocellular carcinoma and was inversely correlated with serum collagenase activity. These results suggest that in this assay condition serum collagenase activity is influenced by the metallo-proteinase tissue inhibitor and thus does not reflect the amount of collagenase in the fibrotic liver. PMID- 8228127 TI - Immunogenicity of rat hepatocytes in vivo: effect of cholestasis-induced changes in major histocompatibility complex expression. AB - Hepatocytes normally express few major histocompatibility complex (MHC) class I and no MHC class II molecules, a phenomenon which could explain their low immunogenicity. However, in pathological situations, such as allograft rejection and cholestasis, hepatocytes strongly express MHC class I molecules and their immunogenicity could be different. The aim of this study was to assess the role of MHC expression on the immunogenicity of hepatocytes in vivo. Hepatocytes were obtained from normal and cholestatic DA rats by whole-liver perfusion with EDTA. Cholestasis was induced by ligation-section of the common bile duct. MHC expression on hepatocytes was assessed by cytofluorimetry after labelling with monoclonal antibodies against MHC class I and class II antigens. The percentage of hepatocytes expressing MHC class I was 9.8 +/- 2.2% in normal rats and 77.2 +/ 3.3% in cholestatic rats (P = 2 x 10(-4)); MHC class II expression was present on 1 +/- 0.5% of normal hepatocytes and 0.4% +/- 0.1% of cholestatic hepatocytes (P > 0.05). Lewis rats received a DA or Wistar-Furth heart allograft 7 days after intravenous injection of 2 x 10(7) hepatocytes from normal or cholestatic DA rats. The DA heart allograft was rejected in 6.3 +/- 0.4 days in Lewis controls, 8.8 +/- 1.1 days (N.S.) in Lewis recipients that received normal DA hepatocytes and 17.6 +/- 3.0 days (P = 2 x 10(-4)) in Lewis recipients that received hepatocytes from cholestatic DA rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228128 TI - Evidence against hepatitis viruses as important causes of severe autoimmune hepatitis in the United States. AB - To determine if the hepatitis viruses are important etiologic factors in autoimmune hepatitis, the clinical, immunoserologic, virologic and HLA phenotypes of 105 of the latter patients were assessed prospectively and compared to 45 patients with chronic viral hepatitis. Patients with autoimmune hepatitis were more often women with higher serum aspartate aminotransferase and immunoglobulin levels than patients with viral disease. Only eight patients (8%) were seropositive for anti-HBc and anti-HBs (four patients) or anti-HCV (four patients) and none with anti-HCV were reactive by second generation immunoassay or recombinant immunoblot assay. Smooth muscle (90 vs. 22%, P < 0.001) and antinuclear (70 vs. 22%, P < 0.001) antibodies were more common in patients with autoimmune hepatitis and the titers more frequently exceeded 1:80 (84 vs. 11%, P < 0.0001). Patients with autoimmune hepatitis were more often positive for HLA B8 (48 vs. 20%, P < 0.01) and DR3 (49 vs. 20%, P < 0.003) and they more frequently had the HLA A1-B8-DR3 phenotype (38 vs. 10%, P < 0.003). Only one of the 120 patients tested for anti-LKM1 was seropositive. We conclude that in an American referral population autoimmune hepatitis usually lacks virologic markers and has a distinctive clinical, immunoserologic and HLA phenotype. Hepatitis viruses are not important immunogenic stimuli for non-organ specific antibodies and they are unlikely to be important causes of this form of autoimmune hepatitis. PMID- 8228129 TI - A prospective study of bacterial infections in patients with cirrhosis. AB - One hundred and seventy hospitalized patients with cirrhosis were included in a prospective and sequential study, to verify the prevalence and most frequent causes of bacterial infection. The differences in clinical and laboratory data between the two groups were analyzed: group I--80 patients who developed bacterial infection and group II--90 patients without bacterial infection. The prevalence or cumulative frequency of the development of bacterial infection during one hospitalization was 47.06%. Among these, the most frequent types of infection were: spontaneous bacterial peritonitis (SBP): 31.07%, urinary tract infection (UTI): 25.24% and pneumonia: 21.37%. Community infections were more frequent (56.25%) than nosocomial infections (32.50%) and they occurred sequentially in 11.25% of the cases. The agents responsible were gram negative bacteria in 72.34% of the cases. Clinical and biochemical parameters in bacterial infection were generally correlated with the severity of liver disease. Child Pugh classification showed a predominance of class C in infected cirrhotic patients compared to non-infected ones. During hospitalization, the mortality rate of group I was 30% whereas in group II it was 5.55% (P = 0.0001). SBP and pneumonia were the most severe types of infection, with high mortality rates, 31.25% and 40.91%, respectively. These results indicate that bacterial infection is a severe complication in the course of cirrhosis. PMID- 8228130 TI - Non-organ specific autoantibodies associated with chronic C virus hepatitis. AB - Recently antibodies to hepatitis C virus were detected in sera of chronic active hepatitis patients, with anti-smooth muscle autoantibodies or with anti liver/kidney microsomal type 1 autoantibodies. As the latter were used to differentiate autoimmune chronic active hepatitis from chronic non-A, non-B virus hepatitis, it was mainly important to discover if autoantibodies were associated with chronic hepatitis C virus infection. The sera of 272 chronic hepatitis C patients were screened by indirect immunofluorescence for non-organ specific autoantibodies. Antinuclear antibodies and anti-smooth muscle autoantibodies were more frequent in chronic hepatitis C patients than in blood donors (n = 100). Anti-liver/kidney microsomal type 1 autoantibodies were not detected in the sera of the blood donors, in the 74 hepatitis B patients or in the 30 alcoholic hepatitis or cirrhotic patients' sera tested as controls. They were detected in 14 chronic hepatitis C patients. These antibodies were compared in immunodiffusion to anti-liver/kidney microsomal type 1 autoantibodies sera obtained from type-2 autoimmune chronic active hepatitis patients and an identity reaction was observed. Chronic hepatitis C patients without or with anti liver/kidney microsomal type 1 autoantibodies, did not differ in age, sex ratio, transaminases and gammaglobulin level, risk factors for hepatitis C virus infection, association with other autoimmune diseases. These patients differed significantly from type-2 autoimmune chronic active hepatitis patients. We conclude that: (i) in some chronic hepatitis C patients the pattern and the titer of autoantibodies may create confusion with an autoimmune chronic active hepatitis; (ii) There is no serological evidence for a hepatitis C virus infection in true type-2 autoimmune chronic active hepatitis. PMID- 8228131 TI - Platelet-activating factor antagonism enhances the liver's recovery from warm ischemia in situ. AB - Hepatic outcome after ischemia is compromised by anoxic injury and by release of oxygen free radicals or other pathological mediators at reperfusion. The platelet activating factor (PAF) is an endogenous lipid-mediator which plays a key role in catalysing various pro-inflammatory processes. In this study the possible influence of platelet-activating factor antagonism was investigated on the integrity of the vascular endothelium, on free radical-mediated peroxidation and on post-ischemic functional outcome of the liver in the rat. Animals under pentobarbital anesthesia were subjected to 60 min of ischemia of the left and median liver lobes followed by 30 min of reperfusion in vivo. Pre-ischemic injection of the platelet-activating factor antagonist BN 52021 resulted in a significant reduction of endothelial enzyme loss into the plasma (purine nucleoside phosphorylase 56.2 vs. 83.1 U/l), and lowered hepatic lipid peroxidation (malondialdehyde 830 vs 932 nmol/g), leading to a significant improvement of postischemic bile flow and higher tissue levels of ATP. This suggests that the platelet-activating factor may play an important role in hepatic ischemia-reperfusion injury and that PAF antagonists like BN 52021 may be useful for clinical treatment. PMID- 8228132 TI - Detection of precore/core-mutant hepatitis B virus genome in patients with acute or fulminant hepatitis without serological markers for recent HBV infection. AB - To confirm the possibility that some hepatitis B virus (HBV) variants do not induce HB s antigen (HBsAg), anti-HB core antibody (anti-HBc) and anti-HBc IgM in a transient infection, polymerase chain reaction (PCR) was performed in 20 patients with acute hepatitis and 7 patients with fulminant hepatitis. Patients were diagnosed with non-A, non-B hepatitis by serological markers at admission. PCR successfully amplified the precore/core gene in 5 (25%) of the patients with acute hepatitis and 2 (29%) of the patients with fulminant hepatitis. Subsequent sequencing revealed frequent mutations including precore-defects in the precore/core gene. PMID- 8228133 TI - Human fibrogenesis and transforming growth factor beta-1. PMID- 8228134 TI - Chronic intrahepatic cholestasis and Sicca syndrome of sarcoidosis. PMID- 8228135 TI - Laparoscopic treatment without cholecystectomy for gallbladder perforation after liver biopsy. PMID- 8228136 TI - The child's perception of the human energy field using Therapeutic Touch. AB - This is a descriptive, exploratory, phenomenological study on children's lived experiences of perceiving the human energy field using Therapeutic Touch (TT). Eleven children, 3 to 9 years of age, willingly participated in the study. The study's findings suggest that children can feel the human energy field with purpose or intent to help, thus supporting TT as an innate potential. PMID- 8228137 TI - Psychoneuroimmunological aspects of nursing: a course sampler. AB - Health care consumers are searching for ways to enhance health. Bookstores are filled with self-help literature about mental, physical, and emotional connections and ways to manage these relationships. In an innovative and challenging course, the authors discuss mind-brain-body interactions and illustrate the application of alternative modalities in nursing practice. PMID- 8228138 TI - Holistic Self-Care Model for permanent weight control. AB - Nearly one fourth of adult women in the United States are currently overweight, despite numerous approaches to weight control. Nurses are confused about appropriate interventions for overweight women who weight cycle--that is, experience repeated periods of weight loss followed by weight regain. Women continue to suffer the negative physical, psychological, and financial consequences of weight cycling, which is related to increased adiposity, an additional important health risk. Moderately successful, long-term weight control treatments are programs that have a combined intervention prescribing both controlled amounts of health food intake and aerobic exercise. Although such a physical program can promote weight loss, the psychological components often are not considered and are the reasons why so many diet/exercise programs fail. The Holistic Self-Care Model combines the successful physical, cognitive, and psychological essentials of healthy approaches for weight reduction as a means of stopping weight cycling and promoting weight control. PMID- 8228139 TI - Hospice and Medicare benefits: overview, issues, and implications. AB - Hospice care is based on a holistic philosophy that considers the physical, psychological, and spiritual dimensions of care for dying patients and their families. It is a family-centered approach and promotes death with dignity. The rapid growth of the hospice movement was in response to dissatisfaction with the care of the terminally ill and demands for death with dignity. Public support for the hospice was so strong that just 8 years after the first hospice was founded, it became a benefit under Medicare. The Medicare hospice benefit emphasizes home care and has shifted the burden of care to families. The restrictive nature of Medicare hospice rules influences who is eligible for hospice care. The Medicare benefit is not congruent with the holistic philosophy of hospice. PMID- 8228140 TI - Life satisfaction, life review, and near-death experiences in the elderly. AB - When 145 senior citizens living independently in the community were surveyed for incidence of near-death experiences (NDEs), 46 of the seniors believed they had had a "close call" with death. Twelve of those reported that they had had an unusual experience during their close call and were subsequently interviewed. Their stories were evaluated using research criteria to document sufficient number of characteristics of an NDE for inclusion in research data and if that set of characteristics of the NDE included a life review component. They were also compared to a non-NDE group on the Life Satisfaction Index, to determine if life review in the context of an NDE helped older people feel more satisfied with life. There was no correlation, because surprisingly, although older people do have NDEs at least as often as other groups, the life review does not seem to be a part of it. So the reasons for scores on the LSI-A are not attributable to the life review. Reasons for this finding are discussed. PMID- 8228141 TI - Patient and family perceptions of restraints. AB - Based on a holistic philosophy that aging persons' most primary need is that of autonomy and freedom, this qualitative descriptive study examined patients' and their family members' perceptions of the restraint procedure. Twenty-five restrained patients were interviewed, and 19 of their family members completed mailed surveys to describe their perceptions about restraints. Results indicated that the majority of patients (52%) expressed negative feelings about being restrained. Family members expressed a wide variety of feelings and were ambivalent about seeing a loved one restrained. Implications are that nursing needs to become committed to the philosophy of autonomy and freedom for elderly persons. PMID- 8228142 TI - Nurses cut health care costs. AB - Nurses are a value-added and cost-savings component of health care, yet others frequently impede nurse efforts. Nurses, coupled with business, can contribute to cutting health care costs by (a) increasing dialogue with business leaders on effective cost-cutting measures across health care, (b) supporting nurse leaders who are capable of administering key community positions, (c) involving whole communities in wellness/health promotion and/or disease prevention programs, (d) encouraging more home health care alternatives; and (e) supporting nurse-related entrepreneurial efforts. PMID- 8228143 TI - New ASTM barrier test methods. American Society for Testing Materials. PMID- 8228144 TI - Guaiac testing of i.v. lines. PMID- 8228145 TI - Port-a-Cath needlestick injuries. PMID- 8228146 TI - Biological indicators for a liquid chemical sterilizer. PMID- 8228147 TI - Legionella disinfection of water distribution systems: principles, problems, and practice. PMID- 8228148 TI - Prevention of Legionella infections in a bone marrow transplant unit: multifaceted approach to decontamination of a water system. AB - OBJECTIVE: To evaluate measures intended to reduce Legionella infections in patients undergoing bone marrow transplantation (BMT). DESIGN: Ongoing clinical and microbiological surveillance for Legionella colonization or infection was undertaken. All neutropenic patients with pulmonary infiltrates and fever unresponsive to broad-spectrum antibiotics were tested for Legionella organisms. SETTING: A 505-bed medical-surgical hospital with a designated BMT unit. PATIENTS: Two hundred twenty-five patients underwent BMT; 201 were treated on a new BMT unit. The incidence of Legionella infections was compared to that seen in an estimated 150 neutropenic patients treated on other units. INTERVENTION: A combined approach to decontamination of a hospital water supply was assessed. This included heating, particulate filtration, ultraviolet sterilization, and monthly pulse hyperchlorination of water supplied to the BMT unit. The incidence of Legionella infections was assessed on the BMT unit and compared with the frequency elsewhere in the hospital. RESULTS: There were only three cases of Legionella pneumonia among 201 patients undergoing transplantation on a new BMT unit. In contrast, 33 cases of Legionella infections were detected from approximately 150 patients treated on general medical floors. CONCLUSION: A multifaceted approach to decontamination of a hospital water system led to a marked reduction in Legionella infections. PMID- 8228149 TI - Infection with methicillin-resistant Staphylococcus aureus among hospital employees. AB - OBJECTIVE: To describe the spectrum of clinical infection caused by methicillin resistant Staphylococcus aureus (MRSA) in healthcare workers. DESIGN: Case series. SETTING: Two Veterans Affairs hospitals in which methicillin-resistant S aureus (MRSA) is endemic. PATIENTS: Five employees presenting to employee health or infectious disease clinic. RESULTS: All employees had had direct exposure to patients colonized with MRSA. Employee infections included cellulitis, impetigo, folliculitis, paronychia, and conjunctivitis. MRSA was isolated from all clinically infected sites and from the anterior nares of two employees. Three employees received a variety of ineffective oral antimicrobials before MRSA was recognized as the causative agent. All infections responded to appropriate therapy. CONCLUSIONS: Employees of hospitals with endemic MRSA may acquire MRSA infection. Presentation in our employees was that of relatively uncomplicated soft tissue infection, but several employees received inappropriate therapy before bacteriologic diagnosis. We recommend that culture and susceptibility testing be obtained prior to institution of therapy when hospital employees present with soft tissue infection. PMID- 8228150 TI - Association between climate and Enterobacter colonization in Swedish neonatal units. AB - OBJECTIVE: To study risk factors for the highly variable local colonization rates with unrelated Enterobacter species strains previously found in 22 Swedish neonatal units (0% to 32.4% of the infants). PATIENTS AND SETTING: The fecal Enterobacter species carriage rates among 953 infants in the 22 special-care neonatal units were correlated with variables related to the ward (size, crowding, staffing, work load, antibiotic usage, level of care, hygienic precautions), and the hospital (temperature of water supplied, geographical location). RESULTS: The average Enterobacter species carriage rate was highest at seven days of age (17% of the infants) and then declined to 3%. Only location of the hospital in an area with warmer climate according to horticultural zone showed an association with Enterobacter species carriage in multivariate analysis (P = 0.005). CONCLUSION: Although Enterobacter species mainly cause nosocomially acquired infections, the occurrence of the organism in special-care neonatal units seemed to be determined more by extrahospital than by intrahospital factors. PMID- 8228151 TI - Outbreak of Candida tropicalis fungemia in a neonatal intensive care unit. AB - OBJECTIVE: To describe an outbreak of Candida tropicalis fungemia in a neonatal intensive care unit (NICU), to evaluate the risk factors associated with this infection and the possible mode of nosocomial transmission. DESIGN: Descriptive and case-control study. PATIENTS AND METHODS: Surveillance cultures were taken from hospitalized patients, personnel, and inanimate objects in the NICU. Six patients with C tropicalis fungemia (cases) were compared with C tropicalis culture-negative patients matched for duration of exposure to the NICU (controls). RESULTS: During a five-month period, C tropicalis was isolated from 29 blood cultures of six premature infants. The same organism also was isolated from fingernail samples taken from the ward housekeeper, who had a mild onychomycosis, and an asymptomatic nurse. Other potential reservoirs of C tropicalis were not identified among all the other infants or in the hospital environment. The six patients with C tropicalis fungemia were more likely to have received a larger number of antibiotics (4.0 versus 1.8, P < 0.001) and to have been subjected to a longer duration of total parenteral nutrition (TPN) therapy (8.5 versus 2.67 days, P = 0.004) than the controls. CONCLUSIONS: The risk of fungemia in this outbreak can be attributed to a larger number of antibiotics and a longer period of TPN administered to the patients. Analysis of events suggests that the outbreak may have been the result of cross-infection between staff and patients. PMID- 8228152 TI - Tetanus and tetanus immunization in long-term care facilities. AB - Although tetanus is rare, elderly residents of LTCFs are at risk for this disease. Assessment of residents' immunization histories, followed by appropriate updating, can prevent tetanus in this population. Many questions, however, remain unanswered. PMID- 8228153 TI - Canada adopts nationwide needlestick surveillance system-EPINet. PMID- 8228154 TI - Infected dental student sued by patients. PMID- 8228155 TI - Barrier testing methods. PMID- 8228156 TI - Prevention of rubella: missed opportunities. PMID- 8228157 TI - A survey of hospital postpartum and postabortion rubella vaccination policies in Los Angeles County, 1992. AB - OBJECTIVE: To determine the proportion of Los Angeles County (LAC) hospitals offering obstetrical services that have postpartum and postabortion rubella vaccination policies. DESIGN: A survey was sent to the infection control practitioners (ICPs) of all operational acute care hospitals (N = 133) in LAC in 1992. A remainder and second survey was mailed to ICPs who did not respond to the first mailing. RESULTS: Of 75 hospitals with obstetrical departments, 56 (75%) responded. Thirty-four (61%) of the 56 respondent hospitals had post-partum rubella vaccination policies. Of the 34 hospitals with policies, 30 (88%) accepted only a written record of rubella seropositivity as proof of immunity, 30 (88%) screened women with unknown immunity status before hospital discharge, and 32 (94%) vaccinated susceptible women before hospital discharge. Of the 32 hospitals that performed induced abortions, only two (6%) provided screening and vaccination services for these women. CONCLUSION: Only 61% of hospitals in LAC offering obstetrical services had postpartum rubella vaccination policies while only minimal screening and vaccination occurred in association with abortion services. Widespread implementation of postabortion screening and vaccination, and more stringent compliance with Advisory Committee on Immunization Practices recommendations for postpartum screening and vaccination in hospitals offering obstetrical services would reduce the number of rubella-susceptible women who have been missed by other prevention strategies. PMID- 8228158 TI - Intervention to discontinue parenteral antimicrobial therapy in hospitalized patients with urinary tract infection, skin and soft tissue infection, or no evident infection. AB - OBJECTIVES: In a previous study, we found that unsolicited recommendations to physicians of medically stable patients with pneumonia to suspend parenteral antimicrobials shortened hospital length of stay (LOS) significantly. In this study, we made similar recommendations to physicians treating patients with different indications for parenteral antimicrobials, to examine the effect on LOS. METHODS: A nurse-interventionist presented randomly assigned physicians with nonconfrontational suggestions to discontinue parenteral antimicrobials by substituting comparable oral antimicrobials or stopping treatment. Patients were being treated for urinary tract infection, skin infection, or no evident infection. Blinded observers evaluated in-hospital and 30-day postdischarge patient courses. Methodologies were identical to the previous study. RESULTS: There were 70 physician-patient episodes (49 intervened episodes, 21 control episodes). In 44 episodes (90%), compliant physicians discontinued parenteral antimicrobials. Compared to a median postrandomization LOS of 2.5 days (range, 0 to 40.5) for 21 patients of control physicians, the corresponding LOS for 44 patients of compliant physicians was two days (range, 0 to 8; P = 1.0), and for five patients of noncompliant physicians, five days (range, 3 to 11; P = 0.04). The combined occurrence of all adverse events detected in this and the previous study was 11% for patients of control physicians, compared to 14% for patients of compliant physicians (P = 0.2), and 19% for patients of noncompliant physicians (P < 0.05). CONCLUSIONS: For patients of compliant physicians hospitalized with urinary tract infection, skin and soft tissue infection, or no evident infection, cessation of parenteral antimicrobials did not significantly shorten LOS, due to brief LOS of patients of control physicians. Patients of noncompliant physicians experienced more adverse events and prolonged LOS. The appropriateness of routine continuous use of parenteral antimicrobials in medically stable inpatients is questioned. PMID- 8228159 TI - Choosing appropriate criteria for tuberculin positivity and conversion in a long term care facility. AB - OBJECTIVES: The Centers for Disease Control and Prevention has issued new criteria for conversion of the tuberculin skin test; in persons over 35 years of age, an increase in induration of at least 15 mm is considered indicative of new tuberculous infection. We reviewed our experience in a tuberculosis control program in a long-term care facility to assess the applicability of the new criteria to our patient population. DESIGN: Retrospective review of seven years of tuberculosis control records and outbreak investigation. SETTING: Long-term care Veterans Affairs hospital. PATIENTS: All patients in the facility between 1985 and June 1992 who received routine admission and annual tuberculin skin testing or who were evaluated for possible exposure to active tuberculosis. A total of 2,342 skin tests were performed. RESULTS: Mean increase in skin test diameter in patients with at least two prior negative tests and known exposure to active tuberculosis was 13.9 +/- 4.7 mm. Frequency distribution histograms of skin test sizes of initial tuberculin testing in the entire population indicated 10 mm induration as a reasonable criterion for initial positivity. CONCLUSIONS: In our long-term care population, an increase in skin test induration of 10 mm may indicate new tuberculous infection. Criteria for skin test conversion derived from ambulatory populations in other geographic areas may not apply in all situations. Prevalence of infection with Mycobacterium tuberculosis and prevalence of skin test reactivity due to nontuberculous mycobacteria are likely to influence the predictive value of criteria for tuberculin conversion in a given population. PMID- 8228160 TI - Inactivation of human immunodeficiency virus by a medical waste disposal process using chlorine dioxide. AB - OBJECTIVE: To study the ability of a medical waste disposal process using chlorine dioxide to inactivate human immunodeficiency virus type 1 (HIV-1). DESIGN: Stock HIV-1 (HTLV-IIIB strain) was treated with chlorine dioxide under the following settings: cell culture medium alone, culture medium with 25% blood, culture medium with medical supplies treated by the Condor machine (Winfield Environmental Corp., Escondido, CA). MT-2 cells in 96-well tissue culture plates were inoculated with serial tenfold dilutions of treated and untreated HIV-1. Cytopathic effect was read on day five, and the TCID50 (50% tissue culture infectious dose) was calculated. RESULTS: Treatment of HIV-1 with chlorine dioxide in culture medium alone resulted in a 5.25 log10 reduction in TCID50. Treatment of HIV-1 with chlorine dioxide in the presence of 25% blood caused a 6.25 log10 reduction in HIV-1 infectivity. Treatment of HIV-1 with chlorine dioxide in the presence of medical supplies treated in the Condor machine resulted in a 4.75 log10 reduction in HIV infectivity. CONCLUSIONS: Chlorine dioxide inactivated HIV-1 in vitro. Chlorine dioxide inactivated HIV-1 in the presence of blood and in the presence of medical supplies under conditions that simulated the conditions existing in the Condor machine. PMID- 8228162 TI - CQI tools: sentinel events, warning, and action limits. PMID- 8228161 TI - Pharmacodynamics of antiretroviral chemotherapy. AB - It has been demonstrated that systemic exposure to antiretroviral agents drives both the therapeutic and the toxic effects seen in clinical trials. Consequently, it is necessary to determine plasma concentrations of drugs and their metabolites (and possibly intracellular concentrations) during the course of Phase 1 and 2 evaluations. It also has been demonstrated that having precise mathematical relationships between exposure and response as well as between exposure and toxicity allows rational selection of drug dose for evaluation in larger Phase 2 and 3 clinical trials. This can be seen with AZT, where the exposure-efficacy relationship is clearly at a plateau by 600 mg/day, while the exposure-toxicity relationship predicts increasing toxicity (and, because of the plateauing of the effect relationship, without better therapeutic response). It can be seen most clearly with ddI, where the exposure-efficacy relationship pointed to longer-term therapy with lower doses ultimately being as efficacious as higher dose therapy. When coupled with the long-term tolerance data for ddI, this led to a prospective prediction of superiority of lower-dose therapy, which was validated later. Finally, some problems with current Phase 1 and 2 trial design have been pointed out, in that bioavailability needs to be recognized explicitly as an important evaluation factor in oral antiretroviral therapy. Perhaps most notably, the importance of initial susceptibility of the virus to the study drug has been suggested by the data of Drusano et al. Clearly, the microbiology of the patient needs to be taken into consideration in the evaluation of Phase 1 and 2 antiretroviral trial results.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228164 TI - Clinton appoints first AIDS czar. PMID- 8228163 TI - C-section deliveries may reduce risk of HIV in newborns. PMID- 8228165 TI - Celadon: an eggshell color mutation in Japanese quail. AB - A new eggshell color mutant was discovered in the Japanese quail. This trait was named "celadon" and was shown to be controlled by an autosomal recessive gene. Therefore, the gene symbol ce was proposed. This gene and an autosomal recessive gene we controlling a white eggshell trait were shown to be nonallelic. Also, ce was shown to be hypostatic to we. A spectrophotometric analysis revealed that the celadon eggshell pigments are protoporphyrin and biliverdin as in the wild-type eggshell, but the protoporphyrin content is much lower than that of the wild type. The biliverdin content was 44% of the wild type. PMID- 8228166 TI - Screening for carriers of the avian cartilage proteoglycan core protein defect, nanomelia, by denaturing gradient gel electrophoresis. AB - The avian mutation, nanomelia, is an autosomal recessive embryonic lethal. Homozygous embryos show hypoplasia of the limbs and a parrot-like beak. Biochemical studies have associated this phenotype with the absence of the major cartilage-specific proteoglycan core protein. In a previous study, a DNA polymorphism was identified at the 3' end of the core protein gene by using denaturing gradient gel electrophoresis. This polymorphism is characterized by three electrophoretic variants (alleles), two of which are associated with the normal core protein gene and one of which segregates with the nanomelia mutation. This study documents that segregation of the latter electrophoretic variant can be used to substitute for progeny testing in the identification of carriers of the nanomelia mutation. Standard progeny phenotype tests were carried out in conjunction with genotype screening for the putative nanomelia-associated electrophoretic variant. We found the genotype data defining the nanomelia core protein gene locus to correlate with progeny phenotype test results. PMID- 8228167 TI - Linkage of the gene for uncoupling protein to esterase-1,2 and haptoglobin in the rat. AB - Genetic analysis of a (BN x ACI)ACI backcross for the biochemical markers Es-1 and Es-2 (linkage group V) and a microsatellite marker of Ucp (chromosome 19) revealed linkage between these three loci. This linkage relationship confirms that in the rat linkage group V genes are located on chromosome 19. Genetic analysis of a (BN x LEW)LEW backcross showed that Hp is linked to Ucp, thus indicating that the Hp locus is also located on chromosome 19. These results substantiate the linkage homology of RNO 19, MMU 8, and HSA 16. PMID- 8228168 TI - Studies in swine on inheritance and variation in expression of small intestinal receptors mediating adhesion of the K88 enteropathogenic Escherichia coli variants. AB - Small intestinal enterocyte preparations from 368 pigs were phenotyped by an in vitro adhesion test using six strains of K88 Escherichia (E.) coli, each expressing one of three K88 fimbrial antigenic variants: K88ab, K88ac, or K88ad. All pigs tested were classified into one of four adhesion brush border phenotypes: I (K88ab-, K88ac-, K88ad-); II (K88ab-, K88ac-, K88ad+); III (K88ab+, K88ac+, K88ad-); or IV (K88ab+, K88ac+, K88ad+). The segregation and adhesion affinity data suggest that there are two adhesion affinity receptors for K88ad+ E. coli: a high affinity (adH) and a low affinity (adL) receptor. The high affinity receptor cosegregates with receptors for K88ab and K88ac fimbrial antigens forming together the phenotype IV; the low affinity receptor is associated with the adhesion phenotype II, and its physiological expression is terminated by 16 weeks of age. In contrast, the K88adH receptor is expressed during the entire life cycle. The presence of a mixed adhesion phenotype, K88adM, assumed to be determined by K88ab(-),ac(-),adL(+)/K88ab(+),ac(+),adH(+) heterozygous genotype, is interpreted as an indication that each of the two types of brush border adhesion for the K88ad antigen is expressed on independent enterocytes. PMID- 8228169 TI - Genetic similarity using DNA fingerprinting in cattle to determine relationship coefficient. AB - The relation between relationship coefficient and genetic similarity was evaluated using DNA fingerprinting in Japanese Black cattle. We proposed a new method for evaluating heterozygous and homozygous minisatellite loci. Genetic similarity by DNA fingerprinting was estimated by two methods--band sharing (BS) and a new evaluation called genetic similarity (GS). Linear regressions of the relationship coefficient (y) on levels of BS (x1) and GS (x2) yielded the equations y = 0.049 + 0.684x1 and y = -0.206 + 1.149x2, with highly significant correlation coefficients of .631 and .813, respectively. It was suggested that DNA fingerprinting could be applied to estimate the relationship coefficient not only in cattle but also in other species. PMID- 8228170 TI - Expressivity of the Manx gene in cats. AB - New genetic data are presented which indicate that the assortment data for the mutant Manx gene, M, does not depart from normal expectation and does not enjoy a selective advantage at some stage of gametogenesis, as has been hypothesized. The variable expression of Manx taillessness is a remarkable and consistent feature of the Manx syndrome, encompassing the posterior skeleton, neural organization, and growth of soft tissues. The expression is partly genetic in origin, and the heritability is estimated to be in the region of h2 = 0.40 +/- 0.11. PMID- 8228171 TI - Analysis of the evolution and genetic diversity within and between Balearic and Iberian cat populations. AB - A study of genic diversity and spatial autocorrelation of some cat populations has been carried out at the microgeographic (cities of Palma Majorca and Ibiza) and the macrogeographic (Balearic Islands and total Spain) levels. In both cases, the cat populations show high levels of gene flow (Nm) and few stochastic effects (e.g., genetic drift). Thus, cat populations are closer to the viewpoint of Mayr (1954)--that is, a species composed of large effective populations very near to panmixia--than to the hypothesis of demic structuration with very little gene flow between populations. The genetic differentiation between cat populations seems to be lower than that found in other vertebrate species. Nevertheless, I have found the existence of significant genetic heterogeneity at both the micro- and macrogeographical levels. At the microgeographical level, the differences in genetic heterogeneity at each of the loci studied within two populations could be caused by the introduction of the diverse mutant alleles into the analyzed populations at different historical times. At the macrogeographical level, the interaction of a strong differential gene flow caused by historical factors (and demographic changes) and, perhaps, natural selection for some loci is a possible explanation. The cat population data are in agreement with the hypothesis of Schwartz and Armitage (1980), who argued that social structures promote and augment gene flow between groups or populations of the same species. Humans seem to be the dominant element that explains the observed high rates of gene flow between cat populations. PMID- 8228172 TI - Blood groups in a marsupial (Monodelphis domestica). AB - We produced alloimmune sera against erythrocytes of the marsupial, Monodelphis domestica. By absorption fractionation, six operationally monospecific blood typing reagents (hemolysins) were prepared. Using segregation analyses, we found that each reagent detects a blood group antigen controlled by an autosomal dominant gene. With the description of these genetic markers, the utility of M. domestica as an animal model is enhanced. PMID- 8228173 TI - Genetics and linkage mapping of Drosophila buzzatii. AB - Of 51 visible mutants isolated from natural or laboratory populations of Drosophila buzzatii, or X-ray induced, 42 have been assigned to chromosomes, and linkage maps have been constructed. About half of the autosomal mutants map to chromosome 2, with only two on chromosome 3 and none on chromosome 4. For the whole repleta group, chromosome 2 also exhibits much greater inversion variability than other chromosomes, which suggests variation among chromosomes in apparent mutability. The chromosomes of D. buzzatii are homologized to those of D. melanogaster and to the standard chromosomal elements of Drosophila. Sequence comparisons for six X chromosome mutant genes, whose homology is reasonably certain, in 13 Drosophila species confirm linkage group conservation but great variation among species in gene order. The linkage group conservation of single copy genes stands in contrast to observed transpositions between elements for tandem repeat genes. PMID- 8228174 TI - Light down--dominant plumage color mutation with homozygous lethality in Japanese quail. AB - A new plumage color mutation in Japanese quail (Coturnix japonica) was named light down (LD) from its external appearance at hatching; it was expressed as an incompletely dominant autosomal gene, for which the symbol L is proposed. In the heterozygous neonates, black stripes on the head and back were narrow and yellowish stripes were wide, creating a lighter external appearance than that of the wild type. One-third of the homozygotes, which had creamy down plumules all over the body, died before hatching. All homozygous chicks allowed to hatch showed abnormal behavior and died within 1 week. The LD adults showed slightly diluted pigmentation all over the body, along with an increased number of transversal bars on the back and wing feathers, as compared with the wild type. PMID- 8228175 TI - Allelism of panda and dotted white plumage genes in Japanese quail. AB - Panda and dotted white mutant plumage colors of Japanese quail (Coturnix japonica) were found to be controlled by the autosomal alleles. Both mutants showed white plumage with colored spots or patches, but they were easily distinguishable by size and location of the colored areas and the color and markings of the pigmented feathers. At hatching also they were easily distinguishable by the position, size, and colors of the spots or patches. The panda chicks possessed large blackish patches on the head, back, tail, cheek, and wing, whereas the dotted white chicks had small light brown spots on the head and/or back. Some dotted white chicks (12%) did not have any brown spots. In a testcross experiment, panda type and dotted white type progeny segregated in 1:0, 3:1, 1:0, and 1:1 ratios in the F1 and F2 generations and in back-crosses to the panda and dotted white parents, respectively, suggesting allelism and dominance of panda over dotted white. The new gene symbol sdw is proposed for the allele controlling the dotted white plumage. PMID- 8228176 TI - Homology within the X chromosomes of Boophilus microplus (Canestrini) and B. annulatus (Say). AB - Chromosome banding patterns of the X chromosomes of Boophilus annulatus (Say) and B. microplus (Canestrini) indicate considerable homology between the two species. All bands of the small X chromosome of B. microplus are represented in the large X chromosome of B. annulatus with the exception of one medially located dark band. A large region of material proximal to the centromere and a small segment located at the telomere of the large X chromosome of B. annulatus are not represented in the small X chromosome of B. microplus. Meiotic abnormalities and the potential differentiation/evolution of the two X chromosomes are discussed. The extensive homology between the two species supports the genic data and linkage analyses previously published for these species. PMID- 8228177 TI - Morphometric evaluation of neurophysin-immunoreactivity in the human brain: pronounced inter-individual variability and evidence for altered staining patterns in schizophrenia. AB - The neurohypophyseal peptides vasopressin and oxytocin have a variety of well documented behavioural effects. Accordingly, terminals and receptors that respectively contain or bind these peptides have been identified throughout the central nervous system (CNS) in experimental animals. In this study we have mapped the distribution of neurophysin I and II-immunoreactive structures in the human CNS. Neurophysins are portions of the precursor proteins for vasopressin and oxytocin, and are found specifically in structures that contain these peptides. In addition, we have quantitatively compared neurophysin neurons and fibers in the brains of individuals with no history of neurologic or psychiatric abnormality, and in brains of patients dying with psychotic illness categorized as schizophrenia. These latter brains were collected prior to the advent of pharmacologic treatment and were available from the Vogt collection. Our findings show wide variations of morphometric values obtained for neurophysin-stained structures in different CNS regions in normal subjects, but provide evidence for abnormal values in certain brain areas in untreated schizophrenia, in the hypothalamic paraventricular nucleus, internal pallidal segment and substantia nigra. These findings suggest that the function of vasopressin and/or oxytocin may be disturbed in these brain regions in schizophrenia. Further investigation will be required to establish whether these differences contribute to, or are a consequence of the disease mechanism. PMID- 8228178 TI - The effect of immunostimulation on thymic innervation in the rat. AB - In order to test whether the thymic innervation shows morphological plasticity in response to thymic functional changes, the density of thymic innervation was determined in sterile control animals and three groups of antigenically stimulated sterile rats. Thymic function was defined from the numbers of mitotic cells present in the thymic cortex, and these were reduced by antigenic stimulation. The innervation was determined separately for the various compartments of the thymus. Nerves were present in the capsule and the septa, but could not be found in either cortex or medulla of the thymic parenchyma. A statistically significant increase in the density of innervation after antigenic stimulation was found for the vessel and the non-vessel innervation of the septa. No significant changes were found in the innervation of the capsule. PMID- 8228179 TI - Energy metabolism and structural factors of long-term grafts in the CNS of rat correlate with graft induced recovery of function. AB - The oxidative energy metabolism and cytoarchitecture of fetal grafts in the neurotoxically damaged hippocampus of adult rats were investigated over a period of 6 months and correlated with adaptive behavioral effects on spatial memory in the Morris water-maze. Three to 6 months postoperatively grafted animals showed significantly lower latency scores in the water-maze compared to lesioned animals with sham grafts. The oxidative metabolism did not show predictory properties for functional efficiency during the first 3 months after grafting. Six months postoperatively the distribution of the cytochrome-c-oxidase reaction product highly correlated with the efficiency of the graft to compensate behavioral deficits as animals with grafts showing a high cytochrome-c-oxidase activity and a high degree of homotypic cytoarchitectural differentiation showed best recovery of functional deficits with only few errors in the maze. There was no significant difference between lesioned and grafted animals during the acquisition phase 1 week after grafting. The histological evaluation revealed that the distribution of cytochrome-c-oxidase was not homogeneous within the grafts. Patches of pyramidal neurons stained with cresylviolet in parallel slices correlated with clusters of high cytochrome-c-oxidase activity. PMID- 8228180 TI - Shifting distribution of autoradiographic label in cone outer segments and its implications for renewal. AB - To clarify how newly-synthesized membrane materials become distributed within the outer segments (OS) of cone photoreceptors during renewal, the retinas of Xenopus laevis were processed for autoradiography at short times after intraocular injection of radioactive glycoprotein precursors (3H-leucine or fucose). Autoradiograms were examined by light microscopy and the labelling density (number of silver grains divided by surface area) was analyzed as a function of COS height. At the first time (2 hours post-injection) that significant label was observed over COS, it was present at all COS heights but not homogeneously distributed. At this time the labelling density in the most basal compartment was significantly greater than that in the rest of the COS, and in some COS the silver grains seemed to form lines of label parallel to the lamellae. At later times, lines were not observed and the labelling density in the most basal compartment had decreased such that at 17.5 hours post-injection the labelling density in the basal and distal regions of COS was similar. At still later times the labelling density in the most basal compartment was significantly less than that in the rest of the COS. Thus, the initial labelling over COS was not diffuse, and during the first 23 hours after injection the location of highest labelling within COS shifted from the base towards the tip. These observations indicate that new membrane does not quickly diffuse so as to become homogeneously distributed throughout the entire COS, nor does it flow exclusively into new lamellae forming at the COS base. The findings are entirely consistent with, and thereby support, an alternative model, namely that new membrane entering COS flows into partial membrane folds that are known from ultrastructural studies of COS to occur both at the extreme base (basal evaginations) and distributed throughout more distal heights (distal invaginations). The diffuse distribution of label observed over COS at later times is used to derive a revised kinetics for COS renewal, which may have implications for cone sensitivity. PMID- 8228181 TI - The neuropathology of schizophrenia: past and present. AB - Historical aspects of neuroanatomical schizophrenia research and recent findings in limbic and basal ganglia structures of schizophrenia are briefly reviewed. Then, results of volume measurements of these brain regions from 26 schizophrenics and 28 controls belonging to our new brain collection are presented. Volumes of the hippocampus, amygdala, stria terminalis and caudate were bilaterally significantly reduced, the internal pallidum showed a trend towards smaller volumes, while brain weight, putamen, nucleus accumbens and external pallidum were unchanged. The results give further evidence of subtle focal brain tissue reductions in schizophrenics. Finally, gaps in our present knowledge in the neuropathology of schizophrenia and some basic problems in this research field are discussed. PMID- 8228182 TI - Regeneration in the peripheral and the central olfactory system: a review of morphological, physiological and behavioral aspects. PMID- 8228183 TI - Persistence of a high degree of structural organization of the telencephalon in brain homotransplants in adult Triturus carnifex (Urodele Amphibians). AB - Brains of adult Urodele Amphibians (Triturus carnifex Laur.) were homoplastically transplanted in conditions of complete morpho-functional isolation and fixed 100 and 365 days after the operation. Results show that in the post-telencephalic regions structural organization disappear in all cases, while, in some cases the telencephalon retain a fairly complex structural organization even after one year. The heterogeneous nature of the histological patterns observed may be attributed to the different size of the matrix areas, besides the different degree of immune response of the host to the graft. In the telencephalic district, where the matrix areas are particularly well developed, undifferentiated elements of these areas would be capable of opposing the degenerative phenomena in the transplants by neoformation of neurons. However, in post-telencephalic districts, where the matrix areas are less developed or absent, it would not be possible to make up for neuronal degeneration and these regions disappear or are formed only by some undifferentiated elements outside the surviving ependyma portions. PMID- 8228184 TI - Bibliography of the current world literature in hypertension. PMID- 8228185 TI - Myocardial cation transport. AB - BACKGROUND: Intracellular cations regulate a variety of functions in myocytes, and abnormalities in ionic homeostatic control have been implicated in several cardiac disease processes. These include cardiac hypertrophy, some of the cardiomyopathies and reperfusion injury following myocardial ischaemia. TOPICS UNDER REVIEW: Current understanding of the sarcolemmal transport mechanisms which generate transmembrane electrochemical gradients for Ca2+, H+ and K+ is reviewed. Both active, ATP-dependent membrane ion transport and secondary active transport are described. The importance of the sarcolemmal Na(+)-K+ pump in maintaining transmembrane gradients for Na+ and K+ is emphasized, and we describe how the electrochemical energy stored in the Na+ gradient generated by the pump is utilized by ion-exchange processes in which a tightly coupled exchange of extracellular Na+ for intracellular Ca2+ or H+ occurs. We also describe cotransport processes in which coupled obligatory transport of Na+, K+ and Cl- occurs in the same direction. Physiological and pharmacological properties of sarcolemmal ion transport mechanisms are reviewed and reference is given to possible clinical implications. PMID- 8228186 TI - The molecular biology of the kallikrein-kinin system: I. General description, nomenclature and the mouse gene family. PMID- 8228187 TI - Sex hormones and hemostatic risk factors for coronary heart disease in men with hypertension. AB - OBJECTIVE AND DESIGN: It has been hypothesized that risk factors for coronary heart disease in men are linked and that the underlying factor linking them may be an alteration in the sex hormone milieu. As a test of this hypothesis, sex hormones and fibrinogen, factor VII and plasminogen activator inhibitor (PAI-1), hemostatic factors recently shown to be risk factors for myocardial infarction, were measured in men with hypertension and in healthy control subjects. RESULTS: The fasting serum testosterone and free testosterone levels were decreased and the plasma factor VII and PAI-1 levels increased in the men with hypertension. CONCLUSION: These findings are consistent with the stated hypothesis. PMID- 8228188 TI - Chronic exogenous hyperinsulinaemia without sugar supplementation: acute salt sensitive hypertension without changes in resting blood pressure. AB - OBJECTIVE: To study the effects of chronic insulin administration without sugar supplementation on blood pressure and response to acute saline loading in normal rats. DESIGN: Comparison of blood pressure, insulin and glucose levels in 24 insulin-treated and 12 control rats on regular rat chow (not supplemented with sugar). METHODS: Sustained-release insulin implants (or sham implantation for the control rats) were administered subcutaneously. The sustained-release insulin implant size was gradually increased. Tail-cuff systolic blood pressure, insulin and glucose were measured twice a week for 8 weeks, after which intra-arterial blood pressure was recorded under resting conditions and 2 h after saline loading in seven insulin-treated and seven control rats. RESULTS: Insulin-treated rats had a 1.2- to twofold increase in insulin without hypoglycaemia, a small but significant increase in glucose levels being found at weeks 6 and 8. When the rats were killed (week 8) triglyceride and fructosamine levels were increased in the insulin-treated rats in comparison with controls. Neither tail-cuff systolic blood pressure nor resting intra-arterial blood pressure differed between the two groups. However, acute saline loading resulted in significantly higher blood pressure in the insulin-treated rats, without altering renal Na+ excretion. CONCLUSIONS: It is possible to produce mild hyperinsulinaemia without hypoglycaemia by gradually increasing subcutaneous sustained-release insulin administration without sugar supplementation. Such hyperinsulinaemia is associated with significantly higher glucose, fructosamine and triglyceride levels, and normal tail-cuff and resting intra-arterial blood pressure. Insulin may induce intolerance to acute volume loading that is not associated with Na+ retention. PMID- 8228189 TI - Effect of chronic and subacute parathyroidectomy on blood pressure and resistance artery contractility in the spontaneously hypertensive rat. AB - OBJECTIVE: Chronic parathyroidectomy (PTX) attenuates blood pressure and increases vascular force generation. The mechanism is not understood. We tested the hypothesis that PTX-induced changes in blood pressure and vessel contractility result from rapid changes in serum parameters induced by PTX, and the hypothesis that the endothelium-derived factors contribute to PTX-induced enhancement of force generation in the spontaneously hypertensive rat (SHR). DESIGN: Chronic PTX or sham surgery was performed on 5-week-old male SHR, and subacute PTX was performed on 13- to 14-week-old SHR. The following experimental parameters were measured 5-6 weeks after chronic and 2 days after subacute PTX: systolic blood pressure, serum Ca2+, 1,25-dihydroxyvitamin D3 (vitD) and mesenteric resistance artery contractility. RESULTS: Chronic PTX significantly reduced blood pressure, body weight and serum Ca2+ compared with sham surgery; subacute PTX reduced only serum Ca2+ and vitD. Compared with sham surgery, the mesenteric resistance artery following chronic PTX showed increased active stress responses to norepinephrine and serotonin that were not associated with changes in free intracellular Ca2+, but were abolished by endothelial denudation. Subacute PTX did not affect the active stress response to norepinephrine or endothelin. Low-dose acetylcholine induced relaxation that was attenuated in the chronic, but not the subacute, PTX subgroup compared with the sham-operated subgroup. High-dose acetylcholine induced contraction that was significantly greater in the chronic, but not subacute, PTX rats than in sham-operated rats. Indomethacin abolished the acetylcholine-induced contraction in all groups, but did not improve the impaired relaxation response of the chronic PTX subgroup. CONCLUSIONS: The PTX-induced decrease in blood pressure and increase in force generation do not result from acute changes initiated by reduction of serum parathyroid hormone, vitD or Ca2+. Moreover, the PTX-induced increase in force generation results from a decrease in the production of a non-cyclo-oxygenase endothelium-dependent relaxing factor. PMID- 8228190 TI - Chronic angiotensin II type 1 receptor antagonism in genetic hypertension: effects on vascular structure and reactivity. AB - OBJECTIVE AND DESIGN: The aim of the study was to assess the role of angiotensin II (Ang II) in the maintenance of cardiovascular hypertrophy and the abnormal vascular amplifier properties in spontaneously hypertensive rats (SHR) with established hypertension. Losartan, a type 1 Ang II receptor antagonist, was administered to SHR and Wistar-Kyoto (WKY) rats, and its effects on blood pressure, cardiac hypertrophy, vascular morphology and hindquarter vascular amplifier properties assessed at the end of treatment and 3 months later. METHODS: Losartan was administered for 6 weeks to 14-week-old SHR (60 mg/kg per day orally). A bio-equivalent dose (20 mg/kg per day orally) was administered to age-matched WKY rats. Systolic blood pressure (SBP) was measured in conscious rats by tail-cuff plethysmography. Morphological changes were assessed both in the heart, from the ratio of the weight of the left ventricular wall plus septum to body weight, and in blood vessels from the medial cross-sectional areas of the abdominal aorta and mesenteric arteries. Vascular amplifier properties were measured by perfusion of the rat hindquarters under conditions of full dilation (papaverine hydrochloride) and incremental constriction with methoxamine hydrochloride. RESULTS: Losartan lowered SBP in SHR to normotensive WKY rat levels during treatment. Left ventricular hypertrophy and aortic cross-sectional area were reduced at the end of treatment to WKY rat levels; mesenteric artery cross-sectional area was reduced to a lesser extent. The abnormal hindquarter vascular amplifier properties of the SHR were normalized by losartan. Three months after treatment ended, SBP had returned to untreated SHR levels. Left ventricular hypertrophy and the abnormal hindquarter vascular amplifier properties had also partially redeveloped. CONCLUSIONS: Our findings support the hypothesis that Ang II contributes to the maintenance of cardiovascular hypertrophy and the abnormal vascular amplifier properties in SHR with established hypertension. However, its role appears to be variable and to depend on the type of vascular bed. Other, pressure-independent, factors may also contribute to vascular hypertrophy. PMID- 8228191 TI - Urinary kallikrein excretion and blood pressure response to angiotensin converting enzyme inhibitors and calcium antagonists in hypertensive patients. AB - OBJECTIVE: To investigate whether the hypotensive effects of angiotensin converting enzyme (ACE) inhibitors in comparison with those of calcium antagonist might be predicted by urinary kallikrein activity, a marker of the activity of the renal kallikrein-kinin system. DESIGN: Seventy-five essential hypertensive patients were randomly assigned to treatment with ACE inhibitors (enalapril or lisinopril 20 mg once a day) or with calcium antagonists (nifedipine 20 mg twice a day or lacidipine 4 mg once a day). Fifty-four had normal (NK) and 21 low (LK) kallikrein activity. Blood pressure was measured after 2 weeks, and 3 and 6 months. Patients whose diagnostic blood pressure, 2 weeks after the first dose, decreased by at least 15 mmHg or was < or = 90 mmHg were defined as responders. The others were defined as non-responders. In non-responders a second drug was added and the patients were not considered for further analysis. METHODS: Urinary kallikrein activity was determined by a spectrophotometric assay using a synthetic chromogenic substrate. RESULTS: After 2 weeks therapy with ACE inhibitors 88% of NK patients were responders, whereas in the LK subgroup 40% were responders, a significant difference between subgroups. For the patients treated with calcium antagonists, conversely, 59% of NK patients were responders in comparison with 82% of the LK subgroup, a significant difference between drug groups. After 3 and 6 months of treatment blood pressure was significantly lower in NK patients treated with ACE inhibitors and in LK patients treated with calcium antagonists. In the NK group on ACE inhibitors the mean arterial pressure after the first dose was significantly related to that observed after 6 months (n = 0.71, P < 0.01). CONCLUSIONS: Our data indicate that urinary kallikrein activity may represent an index to predict the chronic antihypertensive effect not only of ACE inhibition but also of calcium antagonism, and support the concept that the renal kallikrein-kinin system might play some contributory role in modulating the hypotensive action of ACE inhibitors. PMID- 8228192 TI - Blood viscosity and peripheral vascular resistance in patients with untreated essential hypertension. AB - OBJECTIVES: The viscosity of blood is increased in patients with essential hypertension. The aim of the present study was to investigate the importance of the different variables of blood rheology to total peripheral resistance, and to elucidate whether inappropriate regulation of the formation of erythropoietin could be important. DESIGN: Nineteen consecutive patients with untreated essential hypertension were examined and compared with a group of matched healthy volunteers. METHODS: The haemorheologic variables were assessed by rotational viscometry and the haemodynamic variables by bioimpedance cardiography. The serum concentrations of erythropoietin were determined by radioimmunoassay. RESULTS: The whole blood viscosity and peripheral resistance index were elevated in the hypertensive group. The two variables were positively correlated with each other (r = 0.68, P = 0.0015). The plasma viscosity and erythrocyte aggregation tendency were increased and the erythrocyte deformability, measured as fluidity, was decreased in the hypertensive patients. In the male subpopulation (n = 12) the aggregation tendency was positively, and the deformability negatively, correlated with body mass index. The serum concentrations of erythropoietin were equal in the two groups. CONCLUSIONS: The increased total peripheral resistance in patients with essential hypertension may in part be explained by an increased blood viscosity, but the possibility of an opposite cause-effect relationship must also be taken into consideration. The haemorheological abnormalities observed in the present patients cannot be explained by high serum levels of erythropoietin. PMID- 8228193 TI - The effect of intravenous saline loading on plasma levels of brain natriuretic peptide in man. AB - OBJECTIVE: Recent evidence suggests the presence of a dual natriuretic peptide system consisting of atrial natriuretic factor (ANF) from the atrium and brain natriuretic peptide (BNP) from the ventricle. Discrete roles have been postulated for these two natriuretic peptides in the control of circulatory homeostasis. We have therefore compared the release of ANF and BNP in response to an acute saline load to explore a differential pattern of release for the two natriuretic peptides in man. DESIGN: The effects of an acute saline infusion on plasma ANF and BNP concentrations were studied in 10 normal male volunteers. METHODS: Subjects were studied on two study days in the semirecumbent position. An acute intravenous saline infusion (250 ml/min) of 18 ml/kg isotonic sodium chloride potassium chloride solution was administered on one of the two study days. No infusion was administered on the other day as a control. RESULTS: Plasma ANF concentrations increased significantly (P < 0.01) with saline loading without any detectable changes in plasma BNP concentrations up to 60 min following infusion. Heart rate and systolic and diastolic blood pressure were unchanged after saline loading. CONCLUSIONS: We have shown that an acute intravenous saline load causes an increase in plasma ANF concentrations with no detectable increase in plasma BNP at least up to 60 min after the acute saline load in man. These results support the view that the release of ANF and BNP may be regulated differently, especially with regard to the time required for the acute release of each peptide. PMID- 8228194 TI - Neurohormonal and metabolic effects of severe and moderate salt restriction in non-obese normotensive adults. AB - OBJECTIVE: The effects of severe and moderate sodium restriction on blood pressure, neurohormonal activity and serum lipids were studied in non-obese normotensive adults. METHODS: In the first part of the study, 163 subjects were randomly assigned to a diet of 20 or 300 mmol/day sodium for 1 week each. In the second part, 25 subjects were given a diet of 85 or 200 mmol/day sodium for 4 weeks each in random order. RESULTS: After severe salt restriction 19% of the subjects had a significant decline (salt-sensitive group), 15% showed a significant rise (counter-regulator group) and 66% exhibited no change in blood pressure (salt-resistant group). Severe sodium restriction increased plasma renin activity and noradrenaline concentration, as well as serum total and low-density lipoprotein-cholesterol and triglycerides. After correction for haematocrit, only the changes in total and low-density lipoprotein-cholesterol remained significant. The rise in plasma renin activity during salt restriction was steeper in the counter-regulator group than in the other groups, whereas the changes in plasma noradrenaline concentrations were similar in all groups. During moderate salt restriction, plasma renin activity and noradrenaline concentration were significantly increased, but serum lipid concentrations and blood pressure did not change. CONCLUSION: In non-obese normotensive adults, severe and moderate salt restriction stimulates neurohormonal activity. In contrast to severe salt restriction, a moderate reduction in dietary salt intake does not influence blood lipids in normotensive subjects. PMID- 8228195 TI - Control over work pace, job strain and cardiovascular responses in middle-aged men. AB - OBJECTIVE: To assess the effects of control over work pace on cardiovascular stress responses in healthy middle-aged men. DESIGN: The study involved administration in the laboratory of visual matrix and mirror drawing tasks, the pace of tasks being either under the control of the subject (self-paced) or determined externally (externally paced). The work demands in the two conditions were equated. METHOD: Forty men aged 55-65 years were randomly assigned to self paced or externally paced conditions. Blood pressure (recorded continuously using the Finapres), heart rate, cardiac baroreflex sensitivity, salivary cortisol, skin conductance and breathing pattern were monitored at rest, during task performance and at recovery following tasks. RESULTS: Blood pressure and heart rate responses were significantly greater under the externally paced than self paced conditions. The mean increase in blood pressure during the visual matrix task averaged 19.8/9.4 versus 34.1/15.5 mmHg for the self- and externally paced conditions, and 28.2/13.7 versus 41.8/19.5 mmHg in response to mirror drawing, respectively. Performance of the matrix task was less accurate under externally paced than self-paced conditions, but the two groups did not differ in mirror drawing. A reduction in baroreflex sensitivity and increases in cortisol, respiration rate, tidal volume and skin conductance responses were recorded during tasks, but these responses did not distinguish the two groups. Men were divided on the basis of reported job strain associated with their regular work, using the demand-control model. Blood pressure and heart rate responses were particularly pronounced among men reporting high job strain who were allocated to the externally paced condition. CONCLUSIONS: Middle-aged men showed greater stress-related cardiovascular responses when they performed tasks at a pace that they could not control. This pattern may be relevant to the mechanisms through which job strain (high demands associated with low control) influences cardiovascular disease risk. PMID- 8228197 TI - Bibliography of the current world literature in hypertension. PMID- 8228196 TI - Short report: accuracy of the Dinamap portable monitor, model 8100 determined by the British Hypertension Society protocol. PMID- 8228198 TI - Renorenal reflexes in hypertension. PMID- 8228199 TI - The molecular biology of the kallikrein-kinin system: II. The rat gene family. PMID- 8228200 TI - The kidney and essential hypertension: a link to osteoporosis? AB - INTRODUCTION: Abnormalities of calcium metabolism independent of changes in intracellular calcium have been described in patients with essential hypertension. These include increased urinary calcium excretion for a given salt intake, a raised parathyroid hormone level, an increase in urinary cyclic AMP, a tendency for a low serum ionized calcium level, a raised 1,25-dihydroxyvitamin D level and an increased intestinal calcium reabsorption. These changes have been seen as a consequence of a primary renal calcium leak. HYPOTHESIS: We propose that these changes are secondary to a genetic defect in the ability of the kidney to excrete sodium. On the high salt intake in most Western countries (i.e. approximately 170 mmol/day sodium) compensatory mechanisms occur to try to overcome this defect. These compensatory mechanisms are responsible for the rise in blood pressure, but also cause an increase in central blood volume which is the direct cause of the increase in urinary calcium excretion. This causes a slightly negative calcium balance, and the other abnormalities of calcium metabolism can then be seen as a compensatory response to try to restore calcium balance to normal. DISCUSSION: This hypothesis explains the increase in kidney stones in essential hypertension and predicts that hypertensives, in the long term, will be more likely to develop bone demineralization (osteoporosis), as has been demonstrated in some animal models of hypertension. Increases in salt intake will not only cause a further rise in blood pressure, but will also increase urinary calcium excretion and aggravate the other abnormalities. A moderate reduction in salt intake from 170 to 70 mmol/day will lower the blood pressure and will tend to correct the abnormalities of calcium metabolism. It should simultaneously reduce the incidence of renal stones and the long-term risk of bone demineralization. PMID- 8228201 TI - Gene expression of cardiac beta 1-adrenergic receptors during the development of hypertension in spontaneously hypertensive rats. AB - OBJECTIVE: To study adrenergic receptors in the heart tissues of genetically hypertensive rats by evaluating the gene expression and the membrane protein density of beta 1-adrenergic receptors using steady-state messenger RNA (mRNA) levels and a radioligand binding assay, respectively. DESIGN: We compared prehypertensive (5-week-old) and early-hypertensive (13-week-old) spontaneously hypertensive rats (SHR) with age-matched Wistar-Kyoto (WKY) normotensive control rats. METHODS: Polyadenylated RNA was extracted from individual hearts and analysed by the slot-blot technique using a beta 1-adrenergic receptor complementary DNA probe. beta-Adrenergic receptors in myocardial membranes were studied by radioligand binding assay using [125I]-cyanopindolol and the beta 1- and beta 2-selective antagonists CGP 207.12A and ICI 118.551, respectively. RESULTS: beta 1-Adrenergic receptor mRNA levels were slightly higher, and membrane protein density was similar in prehypertensive SHR and age-matched WKY rats. However, both beta 1-adrenergic receptor mRNA levels and beta 1-adrenergic receptor density were lower in the hypertensive SHR than in the control rats. beta 1-Adrenergic receptor mRNA was significantly reduced in older rats of both strains, and this reduction was most evident in the SHR. CONCLUSIONS: The absence of downregulation of beta 1-adrenergic receptors in young SHR, despite published data indicating a higher cardiac noradrenaline turnover than in WKY rats, may suggest that the cardiac hyperadrenergic activity observed in prehypertensive SHR is maintained, at least in part, by the participation of peripheral, postsynaptic component(s) involving beta 1-adrenergic receptor dysregulation. In addition, the present data suggest that the previously reported evidence of an age-related decrease in cardiac beta 1-adrenergic receptors in rats may be determined at the transcriptional level. PMID- 8228202 TI - Dietary salt excess unmasks blunted aldosterone suppression and sodium retention in the stroke-prone phenotype of the spontaneously hypertensive rat. AB - OBJECTIVE: The mechanisms underlying the accelerating effect of high salt intake on the development of vascular injury in the stroke-prone phenotype of spontaneously hypertensive rats (SHRSP) are still not clear. The aim of the present study was to determine whether young SHRSP can excrete a dietary excess of sodium and to characterize the associated hormonal responses. METHODS: Sodium balance and hormonal parameters were studied during a 1-week high-salt diet (4% NaCl) in 6-week-old SHRSP (n = 84), in age-matched spontaneously hypertensive rats (SHR; n = 73) and in normotensive Wistar-Kyoto (WKY) rats (n = 52). RESULTS: Baseline systolic blood pressure (SBP) was similar in SHR and SHRSP and did not change significantly during the high-salt diet. SBP also remained unchanged in WKY rats during the high-salt diet. Despite similar daily sodium intakes in the three groups during the diet, the response of urinary sodium excretion to sodium loading was reduced significantly in SHRSP compared with SHR or WKY rats (F = 4.09, P < 0.001). Plasma renin activity was suppressed significantly by high salt intake in each group to a comparable extent. Plasma aldosterone concentrations were also reduced significantly by sodium loading in all strains. However, a lesser degree of aldosterone suppression was observed in the SHRSP than in both SHR and WKY rats (F = 3.01, P < 0.01). CONCLUSIONS: Young SHRSP show a blunted suppression of plasma aldosterone and a defective sodium excretion during high salt intake. PMID- 8228203 TI - Rabbits with a genetic impairment in baroreceptor reflex sensitivity show abnormal renal haemodynamics and proximal tubular sodium reabsorption in response to a saline infusion. AB - OBJECTIVE: To compare renal haemodynamics and proximal tubular sodium reabsorption (PTSR) in response to an acute intravenous saline infusion in rabbits bred for genetic differences in cardiac baroreflex sensitivity (BRS). Rabbits with low BRS increase their blood pressure significantly on a high-salt diet, in association with an initial delay in sodium excretion. It was hypothesized that this could occur through an impaired baroreflex regulation of renal sympathetic nerve activity. This, in turn, would alter renal blood flow and PTSR. DESIGN: Experiments were performed in two groups of normotensive male rabbits (n = 10 per group), one of which had high BRS (> 5 beats/min per mmHg; group I) and one of which had low BRS (< 4 beats/min per mmHg; group II). Effective renal plasma flow (ERPF) was measured by para-aminohippuric acid clearance, and PTSR by the lithium clearance technique. Sodium, lithium, para aminohippuric acid and glomerular filtration rate were measured from urine samples collected every 30 min (for 90 min) via an indwelling bladder catheter, during a control infusion of glucose (30 mg/ml) NaCl (1.8 mg/ml), and for 2 h after a threefold increase in NaCl. RESULTS: Group I rabbits increased their ERPF by approximately 40%, in response to saline, and doubled their sodium and lithium clearances within the 2 h, but those in group II did not change their cation excretion or their ERPF significantly during this period. Blood pressure did not increase in either group. CONCLUSIONS: A genetic impairment in BRS may be responsible for the inadequate depression of renal sympathetic nerve activity, which results in a failure to increase ERPF and suppress sodium reabsorption in the proximal tubule in response to salt loading. PMID- 8228204 TI - Erythrocyte Na+,K+,Cl- cotransport and kidney function in essential hypertension. AB - OBJECTIVE: To determine whether essential hypertensive patients with high Na+,K+,Cl- cotransport (COT) display alterations of some indices of kidney tubular reabsorption similar to those observed in Milan hypertensive (MHS) rats, which have high COT in both erythrocytes and kidney tubular cells, and hypertension caused by a primary increase of tubular reabsorption. DESIGN: Two sets of experiments were performed. First, renal function in two subgroups of hypertensive patients (one with 'high' and one with 'normal' COT was compared with that in normotensive controls. Secondly, the natriuretic and diuretic effects of a single oral dose of frusemide (25 mg) were analysed in six high- and in six normal- COT hypertensive patients. RESULTS: Compared with normotensives and with normal-COT hypertensives, high-COT hypertensives had lower fractional uric acid excretion and plasma renin activity with similar glomerular filtration rate and urinary sodium and potassium excretion. COT was negatively correlated with fractional uric acid excretion in the essential hypertensive patients but not in the normotensives. The diuretic natriuretic response to frusemide was much higher in high- than in normal-COT hypertensives. CONCLUSION: These results are consistent with the hypothesis that patients with high COT have abnormal renal handling of sodium similar to that observed in MHS rats. PMID- 8228205 TI - The relationship of erythrocyte sodium-lithium countertransport to blood pressure and metabolic abnormalities in a sample of untreated middle-aged male workers. AB - OBJECTIVES: To investigate the relationship between erythrocyte sodium-lithium countertransport and blood pressure in a randomly selected sample of untreated male workers and to evaluate the influence of a set of metabolic abnormalities commonly associated with hypertension on this relationship. DESIGN: A cross sectional investigation of a randomly selected sample of untreated male workers (n = 216, age range 21-59 years) at the Olivetti factory in Pozzuoli, Naples. METHODS: Standardized measurements of anthropometric and metabolic parameters, blood pressure and Na(+)-Li+ countertransport were performed. RESULTS: In a simple linear correlation analysis Na(+)-Li+ countertransport was directly related to plasma triglycerides and uric acid concentrations, body mass index (BMI) and systolic and diastolic blood pressure. Significantly higher values of Na(+)-Li+ countertransport were observed in the two upper quintiles of the serum triglycerides and uric acid distributions, and of the BMI distribution. Na(+)-Li+ countertransport accounted for approximately 2% of the blood pressure variation in this study population, but its contribution to the effect of metabolic covariates was not statistically significant. Hypertensive individuals with one or more metabolic abnormality had a significantly higher mean level of Na(+)-Li+ countertransport than those hypertensives who were free of such alterations. CONCLUSIONS: The results of the present study suggest that a high level of Na(+) Li+ countertransport is more common in those hypertensive individuals who have concomitant metabolic abnormalities than in hypertensives who are free of such abnormalities. PMID- 8228206 TI - Erythrocyte Na(+)-H+ exchange activity in essential hypertensive and obese patients: role of excess body weight. AB - INTRODUCTION: Several authors have described increased Na(+)-H+ exchanger activity in essential hypertension, and an increase in activity of this transport system has also been postulated in situations of hyperinsulinism, such as obesity and essential hypertension. METHODS: We measured Na(+)-H+ exchanger activity in a group of 37 subjects with essential hypertension (18 obese, 19 non-obese), in a group of nine normotensive obese subjects and in a control group of 16 healthy volunteers. Plasma insulin and glucose values during an oral glucose tolerance test were evaluated, together with other variables such as plasma aldosterone, plasma renin activity and plasma potassium. RESULTS: Na(+)-H+ exchanger system activity did not appear to be abnormally raised in the hypertensive subjects, but was significantly increased in the normotensive obese group. Upon dividing the hypertensive subjects into two subgroups on the basis of body mass index, it was noted that, whereas the non-obese hypertensives showed Na(+)-H+ exchanger activity patterns similar to those in controls, the obese hypertensive subjects exhibited increased activity of the transport system. Na(+)-H+ activity correlates with body mass index and shows a significant inverse correlation with plasma potassium. No correlations were found between Na(+)-H+ exchanger activity and the sum of plasma insulin values during the oral glucose tolerance test. CONCLUSION: Na(+)-H+ exchanger overactivity appears to be characteristic in overweight subjects, but would not appear to be a specific feature of essential hypertension. The increased Na(+)-H+ exchanger activity observed in obese subjects may be postulated to be related to the hypermineralocorticoidism characteristic of this condition. PMID- 8228207 TI - Comparison of blood pressure and angiotensin responses to the renin inhibitor Ro 42-5892 and the angiotensin converting enzyme inhibitor enalapril in essential hypertension. AB - OBJECTIVE: To compare the responses of angiotensin II (Ang II) and blood pressure to the renin inhibitor Ro 42-5892 and the angiotensin converting enzyme (ACE) inhibitor enalapril. SUBJECTS: Eight non-sodium-restricted patients with mild-to moderate essential hypertension. DESIGN: A single-blind crossover study. Ro 42 5892 (600 mg orally, once a day) and enalapril (20 mg orally, once a day) were given for 8 days before detailed investigations were carried out. METHODS: Ambulatory blood pressure was measured directly for 24 h by the Oxford technique on three occasions. Off-treatment and on day 8 of treatment with Ro 42-5892 and with enalapril. Ang II was measured by radioimmunoassay after separation by high performance liquid chromatography. RESULTS: Plasma renin activity and Ang II were lowered by 83% [95% confidence interval (CI) 61-105] and 68% (95% CI 49-87), respectively, 0.5-1 h after Ro 42-5892, but after only 3 h values had returned to baseline. Unlike this rapid and short-term suppression of Ang II, the maximal antihypertensive response to Ro 42-5892 (fall in blood pressure 12.9/9.0 mmHg) occurred only after 6 h. Blood pressure returned to baseline after 8 h. In response to enalapril, Ang II was maximally suppressed by 63% (95% CI 32-94) after 2 h and by 83% (95% CI 76-90) after 8 h. Despite early maximal Ang II suppression, the maximal antihypertensive response to enalapril occurred only after 12 h (fall in blood pressure 25.3/16.3 mmHg). With this compound a significant antihypertensive effect was still present 24 h after dosing. CONCLUSIONS: Compared with enalapril at 20 mg once a day, repeated oral administration of a single dose of Ro 42-5892 at 600 mg caused only short-term suppression of Ang II and blood pressure. Suppression of Ang II and reduction in blood pressure were temporally dissociated, both with the ACE inhibitor and the renin inhibitor. This implies that the blood pressure lowering effect of these inhibitors is caused partly by Ang II suppression outside the circulation. PMID- 8228208 TI - Amlodipine and lisinopril in combination for the treatment of essential hypertension: efficacy and predictors of response. AB - OBJECTIVE: We have shown previously that the combination of captopril and nifedipine was effective at peak response but was very short acting. We therefore decided to study the longer-acting angiotensin converting enzyme inhibitor lisinopril and the long-acting calcium antagonist amlodipine, each alone and in combination, in a double-blind, randomized crossover study in which blood pressures were measured at peak and trough. This study provided the opportunity to investigate what parameters in these patients might possibly predict the fall in blood pressure with the individual drugs and with the combination. METHODS: Fifteen patients with essential hypertension (eight male, 10 Caucasian; mean age 53 years) were studied. After 1 month observation on no treatment they were entered into a single-blind run-in of placebo given once a day for 1 month. Patients were then allocated randomly to amlodipine (5 mg once a day), lisinopril (10 mg once a day) or their combination (once a day) for 1 month in a double blind crossover study. All patients were studied on their usual diet and no dietary advice was given. Blood pressure was measured by semi-automatic ultrasound sphygmomanometer both 24 h and 6 h (trough and peak) after the last dose. RESULTS: During the crossover part of the study there was a significant additional blood pressure-lowering effect (at trough) of the combination compared with either amlodipine or lisinopril alone. Similar results were observed for the blood pressures at peak. The fall in blood pressure with lisinopril was related to baseline plasma renin activity, whereas when amlodipine was given, either alone or in combination, the fall in blood pressure was independent of baseline renin activity. The Blacks (n = 5) appeared not to respond as well to lisinopril as the Caucasians (n = 10). Finally, the blood pressure response to amlodipine tended to be associated with the severity of hypertension. CONCLUSIONS: The results of the present study indicate that: amlodipine and lisinopril in combination have a marked additional effect on blood pressure compared with either given as a monotherapy; their potentiation of action is long-acting; Black patients tend not to respond to the monotherapy with lisinopril as well as Caucasian patients, although they respond similarly to the combination; the response to amlodipine tends to be greater the higher the initial blood pressure; and, finally, the response to lisinopril is greater the higher the plasma renin activity. PMID- 8228209 TI - Biochemical and neurohormonal responses to the introduction of a lacto ovovegetarian diet. AB - OBJECTIVE: To assess the mechanisms of the blood pressure-lowering effect of a vegetarian diet in the early and later stages of dietary intervention. DESIGN: After 2 weeks without intervention (baseline), 20 normotensive men were matched for age and body mass index and randomly allocated to an omnivorous (control) or a lacto-ovovegetarian diet for 6 weeks in a parallel trial. METHODS: Ambulatory blood pressures were recorded between 0800 and 1700 h on alternate days during the first week of intervention, twice in the second week and weekly thereafter. Blood samples collected after a standard breakfast were analysed for plasma noradrenaline, adrenaline, atrial natriuretic peptide (ANP), renin, aldosterone, glucose and insulin. Factor and multiple regression analyses were used to assess the association among neurohormonal factors, blood pressures and diet. Results were analysed for the first week of the diet and for the entire 6 weeks. RESULTS: Ambulatory blood pressures at work were lower on the vegetarian diet than in the controls. This blood pressure decrease was associated with a factor representing lower plasma catecholamine and renin activity levels throughout the study, and a factor representing reduced plasma glucose and insulin levels in week 1 of intervention only. Plasma ANP levels were significantly higher during week 1 of the vegetarian diet. CONCLUSIONS: A blood pressure lowering effect of a vegetarian diet during normal working activity was shown. Using factors derived from the biochemical variables, results were in keeping with the hypothesis that these effects may be mediated by reduced sympatho-adrenal activity consequent to altered glucose and insulin handling. An early increase in plasma ANP may contribute to the blood pressure reduction. PMID- 8228210 TI - Haptoglobin polymorphism and complications in established essential arterial hypertension. AB - OBJECTIVE: Salt sensitivity and the magnitude of systolic blood pressure have been linked to haptoglobin (Hp) polymorphism in normotensives. The aim of the present study was to investigate the indices of hypertension, the severity of complications and the occurrence of coronary and peripheral artery disease for the various haptoglobin phenotypes and their relation to the therapeutic needs (number and class of drugs) of established arterial hypertensives. DESIGN: Haptoglobin polymorphism was studied in 302 Caucasians with established essential arterial hypertension who had been treated for at least 1 year. METHODS: Haptoglobin polymorphism was studied using starch-gel electrophoresis of haemoglobin-supplemented serum. RESULTS: The relative allele frequencies of Hp 1 and Hp 2 (0.036 and 0.640, respectively) in established hypertensives were comparable with those of the control population. Logistic regression analysis confirmed that Hp 2-2 contributes to the therapeutic needs in hypertension. The most important factors determining therapeutic needs were coronary artery disease, Hp 2-2 phenotype, body mass index (BMI) and left ventricular hypertrophy. Although no contributive effect of serum haptoglobin concentration could be derived from the logistic regression approach, analysis of serum haptoglobin concentration demonstrated a concentration-related effect on therapeutic needs for the Hp 2-2 phenotype only. CONCLUSIONS: The present study suggests that hypertensives with an Hp 2-2 phenotype need more complex combinations of antihypertensive drugs to reduce blood pressure to the same level. The hypertensive patient carrying Hp 2-2 is more likely to accumulate atherosclerotic lesions of the coronary or peripheral arteries, despite comparable lipid levels, smoking habits and BMI. Hp 1-1 patients are characterized by a younger age at diagnosis and a lower complication rate. In view of the greater therapeutic needs and the higher complication rate, Hp 2-2 hypertensives need more careful follow-up. PMID- 8228211 TI - Twenty-four-hour ambulatory blood pressure measurement in a primigravid population. AB - OBJECTIVE: To establish the profiles of 24-h non-invasive ambulatory blood pressure measurement (ABPM) during the trimesters of pregnancy and the puerperium in normotensive healthy primigravidae. DESIGN: A prospective study in which 24-h ABPM was performed on five occasions in each subject: in the first trimester between 9 and 16 weeks' gestation; in the second trimester between 18 and 24 weeks; in the third trimester between 26 and 32 weeks and between 33 and 40 weeks; and finally at 6 weeks post partum. METHOD: One hundred and six Caucasian primigravid women who were normotensive at their first booking visit were recruited consecutively from the antenatal clinic and had 24-h ABPM performed with the SpaceLabs 90207 ambulatory system. RESULTS: Of the 106 women recruited, 98 completed 24-h ABPM on four of the five measurement occasions. Four women delivered prematurely before 33 weeks' gestation, thereby missing one ABPM measurement. Changes during pregnancy and the puerperium were assessed against the ABPM performed in the first trimester. There was no difference for daytime or night-time systolic blood pressure between 9 and 33 weeks, but it rose significantly from 33 to 40 weeks. At 6 weeks post partum, systolic blood pressure was not significantly different from the daytime pressure in the first trimester ABPM but was raised significantly at night. Diastolic blood pressure decreased significantly between 18 and 24 weeks for both daytime and night-time. From 33 to 40 weeks it increased in parallel with systolic blood pressure, and at 6 weeks post partum it was raised significantly compared with first-trimester values for daytime and night-time. The nocturnal fall in blood pressure was preserved throughout pregnancy with a significant difference between daytime and night-time measurements present on all measurement occasions for systolic, diastolic and mean blood pressures and heart rate. There were significant differences between daytime ABPM and clinic blood pressure for both systolic and diastolic blood pressure up to 33 weeks. From 33 weeks until 6 weeks post partum there was no significant difference between daytime ambulatory and clinic blood pressures. CONCLUSION: This study provides reference values for ABPM in healthy primigravidae with generally uncomplicated pregnancies. PMID- 8228212 TI - Evaluation of organ damage in hypertension. AB - INTRODUCTION: This document is the result of discussions held during a Scientific Workshop in Brescia, Italy, where the numerous methods, old and new, for evaluation of organ damage in hypertension were considered. Not all of the methods can be recommended as routine diagnostic procedures for every hypertensive subject. Current routine diagnosis of organ damage by clinic blood pressure, retinal appearances, the electrocardiogram, serum creatinine estimation and urinary analysis will probably suffice; of the more refined methods, only the echocardiogram is likely to be often included in the examination of hypertensive patients. All of the other techniques will be useful in the diagnostic approach to patients with particular problems, or for the purposes of special investigation. RECOMMENDATIONS: It is recommended that the validity of the methods, the clinical relevance of the abnormalities (including left ventricular hypertrophy, reduced coronary reserve, arterial changes in the eye grounds, reduced large artery compliance and microalbuminuria) and the potential benefits of therapeutically reverting these alterations should be assessed by including organ damage evaluation in future epidemiological and interventional trials in hypertension. PMID- 8228213 TI - Cross-linking of HLA class II antigens modulates the release of tumor necrosis factor-alpha by the EBV-B lymphoblastoid cell line JY. AB - In addition to their functional role as peptide-binding proteins HLA class II Ag can also act as signal-transducing molecules. The present study showed that cross linking of HLA class II Ag by the anti-HLA-DR mAb L243 or by the anti-HLA-DR,-DP mAb IVA12 significantly (p < 0.05) increased the release of TNF-alpha by the EBV B lymphoblastoid cell line JY. In contrast, the anti-HLA-DR mAb 2.06 or the superantigens staphylococcal exotoxin toxic shock syndrome toxin-1 and staphylococcal enterotoxin B that bind to HLA-DR,-DQ Ag did not affect the release of TNF-alpha by JY cells. The accumulation of TNF-alpha in the culture medium of JY cells peaked at 24 h, decreased thereafter, and was found to be dependent on the dose of mAb L243 or mAb IVA12 used to cross-link HLA class II Ag. mAb L243 or staphylococcal exotoxin toxic shock syndrome toxin-1 enhanced the spontaneous homotypic aggregation of JY cells and mediated a dose-dependent inhibition of JY cell proliferation. These phenomena were not mediated by TNF alpha released in response to cross-linking of HLA class II Ag; polyclonal anti TNF-alpha neutralizing antibody did not affect JY cell aggregation and the inhibition of JY cell proliferation mediated by mAb L243. In contrast, TNF-alpha secreted by JY cells enhanced a nuclear factor-kB-like activity through the binding to the 75-kDa TNF-alpha receptor. These results demonstrate an additional role of HLA class II Ag as signal-transducing molecules regulating the production of bioactive TNF-alpha by EBV-B cells. The release of TNF-alpha after the triggering of HLA class II molecules could be relevant to different aspects of B cell biology and might play a role in the pathogenesis of human diseases in which antibodies cross-reactive to HLA class II Ag have been identified. PMID- 8228214 TI - Regulation of CD4 and CD8 expression on mouse T cells. Active removal from the cell surface by two mechanisms. AB - Phorbol esters are known to induce a loss of CD4 from the surface of mouse and human T cells, presumably through activation of protein kinase C. Here we describe additional, calcium-dependent processes that remove CD4 and CD8 from the surface of T cells and thymocytes, and that differ from the protein kinase C mediated effect in that they require the expression of new gene products. Whereas PMA causes the disappearance of almost all CD4 from the surface of mouse CD4+8+ thymocytes, it induces only a partial (approximately 60%) and transient (10 to 12 h) loss of CD4 from the surface of mouse peripheral T cells, with no effect on CD8 expression. When T cells are exposed to a combination of PMA and the calcium ionophore, ionomycin (Cal), surface CD4 virtually disappears for a period of at least 24 h, and CD8 expression is also diminished. This additional, calcium dependent effect, on both CD4 and CD8 expression, is abrogated by either cycloheximide or actinomycin D, and so depends on new RNA and protein synthesis. There appear, therefore, to be two distinct mechanisms for the removal of CD4 and CD8 from mouse peripheral T cells: one induced by PMA alone, the second by the combination of PMA and Cal. The second mechanism, but not the first, depends on the expression of new gene products. In contrast to mouse peripheral T cells, mouse thymocytes and human peripheral blood T cells respond to PMA alone with virtually a complete loss of surface CD4, and partial loss of CD8 expression. The addition of Cal has no synergistic effect on CD4 expression in either of these populations, but augments the loss of CD8 in mouse thymocytes. PMID- 8228215 TI - Contrasting effects of transforming growth factor-beta and IL-1 on the regulation of plasminogen activator inhibitors in human synovial fibroblasts. AB - TGF-beta increased in a dose-dependent manner the production of plasminogen activator inhibitor-1 (PAI-1) in cultured human synovial fibroblast-like cells, as measured by ELISA. Significant increases in PAI-1 were first detected in cell supernatants within 4 h after cytokine addition. Increases were also observed in PAI-1 mRNA expression. IL-1 suppressed these increases in PAI-1 Ag and mRNA. In contrast, when PAI-2 levels were measured by ELISA, TGF-beta did not raise them but inhibited slightly the enhancement caused by IL-1 of PAI-2 Ag and mRNA. Therefore TGF-beta selectively stimulates the formation of one PAI; TGF-beta and IL-1 have opposing effects on PAI-1 and PAI-2 synthesis in the synovial cells. These findings are proposed to help define the control of fibrinolysis and tissue remodeling in the rheumatoid synovium. PMID- 8228216 TI - Evidence that peritoneal exudate cells cultured with eye-derived fluids are the proximate antigen-presenting cells in immune deviation of the ocular type. AB - Immune privilege in the anterior chamber of the eye results in part from a selective deficit in delayed hypersensitivity that is elicited by antigenic materials placed in this unique tissue site. This distinctive systemic immune response to intraocular Ag (termed anterior chamber-associated immune deviation, ACAID) is fashioned by indigenous, intraocular bone marrow-derived cells that capture Ag within the anterior chamber and carry an Ag-specific ACAID-inducing signal via the blood directly to the spleen. An identical form of immune deviation can be evoked by the i.v. injection of peritoneal exudate cells (PEC) pulsed in vitro with soluble Ag in the presence of transforming growth factor beta (TGF-beta). To determine whether eye-derived cells present Ag directly to responding splenic T cells or merely serve as vehicles to deliver Ag to the spleen, we have conducted MHC restriction experiments with PEC donors and recipients selected to differ at loci dictating MHC and/or minor histocompatibility Ag. When PEC were pulsed in vitro with soluble Ag (BSA) in the presence of culture fluid containing TGF-beta and injected into recipients with which they shared either class I or class II MHC molecules, BSA-specific ACAID was induced. By contrast, PEC pulsed with BSA and TGF-beta-containing culture fluid failed to induce ACAID when the cells were injected into recipients who were completely histoincompatible or were identical with the injected cells only at non-MHC loci. Using MHC class I-deficient transgenic mice, it was determined that intraocular injection of BSA failed to induce ACAID, and that class I deficient PEC were incapable of inducing either BSA-specific ACAID or splenic regulatory T cells that suppress BSA-specific delayed hypersensitivity. We conclude that cells that carry an ACAID-inducing signal to the spleen are: 1) restricted in their ability to induce ACAID by MHC-encoded molecules, 2) these cells are the proximate APC ACAID, and 3) class I MHC molecules play a central role in presenting exogenous protein Ag to splenic T cells in ACAID. PMID- 8228217 TI - Effects of purified dietary n-3 ethyl esters on murine T lymphocyte function. AB - To precisely determine the biologic role of fish oil-derived constituents (n-3 polyunsaturated fatty acids), it is imperative that information on highly purified n-3 polyunsaturated fatty acids be evolved. Therefore, we studied the effects of a low dose, short term dietary supplementation with highly purified (n 3) ethyl esters with regard to murine T lymphocyte function. A 10-day dietary supplementation with low dose, highly purified (n-3) fatty acid ethyl esters was examined for effects on murine splenic lymphocyte function and membrane composition. Mice were fed diets containing either 3% safflower oil (SAF) ethyl esters, 2% SAF plus 1% eicosapentaenoic acid ethyl esters (EPA) (99% pure), or 2% SAF plus 1% docosahexaenoic acid ethyl esters (DHA) (97% pure). Fatty acid analysis of the lymphocyte membranes showed that membrane compositions of the EPA and DHA-fed mice were subsequently enriched with these (n-3) fatty acids. Con A induced lymphoproliferative assays (Con A, 5 and 10 micrograms/ml) revealed that splenic lymphocytes from the EPA group had significantly higher mitogenic responses relative to lymphocytes from the DHA or SAF groups (p < 0.05). In contrast, DHA splenocytes had the highest level of 3H-labeled diradylglycerol as a percentage of total lipid. Macrophage-lymphocyte co-cultures demonstrated that the dietary effect on proliferation in response to Con A was influenced by lymphocyte source, but apparently not by macrophage source. In addition, cells from Mycobacterium bovis vaccine-vaccinated mice were placed in culture with purified protein derivative (PPD) (10, 20, and 40 micrograms/ml), as well as Con A (5 and 10 micrograms/ml) to allow comparison between mitogenic and antigenic stimulation. No statistically significant differences (p > 0.05) existed between the diet groups upon PPD stimulation. However, lymphocytes from the DHA group exhibited significantly higher proliferative responses to Con A than cells from the EPA and SAF groups (p < 0.05). M. bovis vaccine-immunized EPA and DHA fed mice also exhibited significantly reduced delayed-type hypersensitive reactivity to footpad testing with PPD in comparison to that demonstrated by SAF-fed mice (p < 0.05). EPA-fed mice demonstrated the most decreased response. Overall, this study demonstrates for the first time that: 1) low dose, short term dietary supplementation with highly purified EPA or DHA can modulate select functional and transmembrane signaling responses of murine splenic lymphocytes; and 2) that these dietary agents have differing effects on mitogen vs Ag receptors. PMID- 8228218 TI - Early mobilization of Ca2+ is not required for glucocorticoid-induced apoptosis in thymocytes. AB - Contradictory results have been reported on the question of the role of Ca2+ in glucocorticoid-induced apoptosis in thymocytes. To resolve this problem, we investigated the effect of dexamethasone, a synthetic glucocorticoid, on intracellular Ca2+ concentration ([Ca2+]i), by microscopic fluorometry that enables us to monitor real-time [Ca2+]i of cells loaded with fura-2, a fluorescent Ca2+ indicator, on a single cell basis. The results indicated that dexamethasone does not induce an increase in [Ca2+]i above control level both in murine and rat thymocytes at least for 1 h after the start of the culture. We also investigated whether the depletion of extracellular Ca2+ with EGTA or buffering intracellular Ca2+ with quin-2/AM inhibited glucocorticoid-induced apoptosis as reported on rat thymocytes. Dexamethasone-induced apoptosis in both murine and rat thymocytes, however, was not inhibited by EGTA. High concentrations (25 microM and over) of quin-2/AM inhibited DNA fragmentation, but failed to inhibit cytolysis. Calmodulin inhibitors, trifluoperazine and calmidazolium, also inhibited DNA fragmentation as reported, although they markedly enhanced cytolysis. Therefore, glucocorticoid-induced death is not inhibited by quin-2/AM or calmodulin inhibitors. Furthermore, we have previously found that a proper combination of the calcium ionophore, ionomycin, and the protein kinase activator, PMA, inhibits corticosterone-induced apoptosis. These results suggest that an early increase in [Ca2+]i is neither induced by glucocorticoids nor responsible for glucocorticoid-induced apoptosis in thymocytes. PMID- 8228219 TI - cAMP analogs prevent activation-induced apoptosis of T cell hybridomas. AB - Activation of T cell hybridomas through their TCR leads to secretion of IL-2, inhibition of proliferation, and apoptosis. The identification of various inhibitors that prevent activation-induced T cell death (AICD) has helped identify several essential events in apoptosis. For example, inhibition of AICD by dexamethasone indicates a connection between these two programmed death pathways. In this study, we have investigated the interaction between the cAMP signal transduction pathway and the activation- or glucocorticoid-induced cell death. cAMP induced DNA fragmentation in thymocytes. T cell hybridomas displayed different sensitivity to cAMP. Regardless of its cAMP sensitivity, programmed cell death promoted by anti-CD3 or Ag in hybridoma was prevented by the presence of cAMP analogs. In contrast, cAMP had no effect on glucocorticoid-induced T cell death. The inhibitory effect of cAMP on AICD was unlikely to be due to quenching of T cell activation signals, because cAMP added 1 h after T cell activation could still prevent cell death. In addition, the increased binding of AP-1, NF AT, and NF-kappa B during T cell activation was not significantly affected by cAMP. The presence of the inhibitory cAMP-mediated signals, together with the glucocorticoid-induced pathway, suggest there are at least two distinct mechanisms regulating AICD in immature lymphocytes. PMID- 8228220 TI - Alloantigen presentation by individual clones of mouse splenic macrophages. Selective expression of IL-1 alpha in response to CD8+ T cell-derived IFN-gamma defines the alloantigen-presenting phenotype. AB - Approximately one-third of mouse splenic macrophage (M theta) progenitors yield progeny that constitutively present MHC class I alloantigen to naive T cells, a response that is restricted to CD8+ T cells and is elicited in a CD4+ Th cell independent manner. In addition, both the alloantigen-presenting (alloAP+) and nonpresenting (alloAP-) M theta subsets constitutively express similar levels of MHC class I molecules, and their alloAP phenotypes are unaffected by IFN-gamma, which enhances the expression of both class I and II MHC molecules. We therefore postulated the restricted expression of costimulator molecules to account for the alloAP+ phenotype. Using cytokine-specific antibodies, recombinant mouse cytokines, and polymerase chain reaction analyses (to detect specific cytokine mRNA transcripts), we identified the putative costimulators as IL-1 alpha, IL-6, and TNF-alpha. TNF-alpha transcripts were present in both the alloAP+ and alloAP- M theta subsets, but IL-1 alpha and IL-6 were not constitutively expressed by the alloAP+ subset of M theta; rather, they were induced by IFN-gamma, which was released from naive CD8+ T cells only during coculture with alloAP+ M theta. Although IFN-gamma induced IL-6 gene transcription in both alloAP+ and alloAP- M theta subsets, it induced IL-1 alpha transcripts only in the alloAP+ subset. Finally, CD8+ T cells exposed to alloAP- M theta were unresponsive when subsequently cultured with alloAP+ M theta. We conclude that the ability of some M theta to elicit IFN-gamma from CD8+ T cells and to respond to this cytokine by producing IL-1 alpha defines the alloAP phenotype of the cell population, and that alloAP- M theta induce a state of alloantigen-specific tolerance in naive CD8+ T cells. PMID- 8228221 TI - Lymphocytes infiltrating rat cardiac allografts express a limited repertoire of T cell receptor V beta genes. AB - The T cell response to many self-MHC-restricted nominal Ag involves limited use of the TCR repertoire. The status of the TCR repertoire in allogeneic responses remains unclear. In this report, we have studied the TCR V beta gene repertoire involved in the rejection of cardiac allografts disparate for major and minor histocompatibility Ag in rats. Graft-infiltrating lymphocytes (GIL) were isolated from rejecting heart allografts and analyzed for the expression of the V beta repertoire using a cDNA library and a semiquantitative polymerase chain reaction (PCR). We report here that GIL isolated at early stages of the rejection reaction preferentially use the V beta 4 gene. First, V beta 4 comprised 36.4% (8/22) of randomly sequenced cDNA clones isolated from a TCR-beta chain-specific cDNA library established from GIL harvested 3 days posttransplantation. The V beta 4 gene in these clones was found in conjunction with several different J beta and N regions, suggesting a dominant role for the V beta 4 encoded domains in the recognition of allograft Ag. Second, the V beta 4 message comprised 56.6 to 65.7% of the transcripts expressed by the 20 rat V beta genes in three T cell lines established from GIL isolated 2 days posttransplantation. Third, fresh, unmanipulated GIL harvested at days 2 and 3 posttransplantation predominantly expressed the V beta 4 gene. Fourth, the expression of V beta 4 in naive splenocytes constituted only 5.4% of the V beta detected, suggesting that the predominant use of the V beta 4 gene by GIL was not a consequence of its high level of expression in the periphery. The limited use of the TCR repertoire in allograft rejection may provide the opportunity to interrupt the rejection process and induce donor-specific tolerance by targeting a select population of T cells for inactivation or elimination. PMID- 8228222 TI - Natural killer cells induce activated murine B cells to secrete Ig. AB - We previously demonstrated that dextran-conjugated anti-IgD antibodies (alpha delta-dex) induce proliferation of small, B cell-enriched murine spleen cells (Be cells), and in the presence of IL-2, stimulate Ig secretion in vitro. We have shown that alpha delta-dex-stimulated B cells provide an in vitro model for studying B cell activation by T cell-independent type 2 (TI-2) Ag, as exemplified by the bacterial polysaccharides. We now show that highly purified resting B cells, obtained by electronic cell sorting (Bsp cells), fail to secrete Ig in the presence of alpha delta-dex + IL-2. The alpha delta-dex + IL-2-induced Ig secretory response of Bsp cells is restored upon addition of splenic non-B, non-T cells or a pure population of in vitro-generated NK cells. Similarly, pretreatment of Be cells with anti-AsGm-1 plus complement inhibits Ig secretion in response to alpha delta-dex + IL-2. An IL-2-induced NK cell supernatant (NKSN) is equally potent at stimulating Ig secretion by alpha delta-dex-activated Bsp cells, indicating that cell contact between Bsp and activated NK cells is not required for this effect. IL-2 stimulates not only NK cells, but B cells as well, since addition of anti-IL-2 + anti-IL-2R antibodies to Bsp cell cultures, in the presence of alpha delta-dex + NKSN, inhibits Ig secretion. These data describe a novel animal model for NK cell-induced B cell maturation to Ig secretion and suggest a pathway for Ig production in response to T1-2 Ag. PMID- 8228223 TI - Cloning of mouse Ox40: a T cell activation marker that may mediate T-B cell interactions. AB - A cDNA library was prepared from the murine Th cell line Th2 D.10 and used to clone the murine homologue of Ox40 by polymerase chain reaction. Comparison of the mouse sequence with the rat revealed greater than 90% homology between the two sequences at both the DNA and protein level. Northern blot analysis found that, as in the rat, Ox40 expression appears to be restricted to activated T cells. A chimeric receptor globulin was prepared to include the mouse Ox40 extracellular domain coupled to the hinge-CH2-CH3 domains of human IgG1 (Ox40 Ig). This soluble form of the molecular was then used to identify cells bearing a ligand for Ox40. FACS analysis revealed that Ox40-Ig bound to a subset of peritoneal B cells as well as to a fraction of LPS-activated splenic B cells. Immunostaining of spleen sections using an Ag-specific conjugate and Ox40-Ig found a significant proportion of antibody-forming cells co-stained with Ox40-Ig. Immunoprecipitation of cell-surface radiolabeled peritoneal B cells suggests a specific interaction with a protein of 70 kDa. PMID- 8228224 TI - Length and sequence requirements of the cytoplasmic domain of the A beta molecule for class II-mediated B cell signaling. AB - APC use class II molecules of the MHC to present peptide Ag to Th cells. Interaction of the TCR and CD4 with the class II-peptide complex, together with co-stimulatory signals provided by the APC, activates the T cell. B lymphocytes express class II molecules and can also be induced to express co-stimulatory molecules, allowing them to act as APC to Th cells. In addition to T cell activation, class II binding by T cells has been shown to result in the transmission of signals to B cells. Signal transduction via MHC class II has been well documented in B cells of both mice and humans and is implicated in the processes of cellular adhesion, Ag presentation, and Ag-dependent B cell activation. The regions of the class II MHC molecule which are involved in signal transduction to the B cell are not clearly defined. However, previous studies have suggested that the beta chain of the alpha beta heterodimer has a predominant role in B cell signaling. To examine the role of the cytoplasmic domain of this molecule in class II-mediated signaling to a mouse B cell clone, we have prepared and analyzed a set of subclones expressing sequentially truncated forms of A beta b. Our results demonstrate that only the 8 membrane proximal amino acids of the cytoplasmic domain are required for signaling. However, specific conserved amino acids within this minimal length are required for successful signal transduction; length alone is not sufficient. Examination of the signaling ability of these truncated beta chains suggests that conserved residues at positions 227 and 228 of the cytoplasmic domain may have particularly important effects on signal transduction. A beta b chains from which the entire cytoplasmic domain have been removed are still capable of transmitting a detectable, although reduced, signal to B cells. Thus, the transmembrane and/or extracellular domains may also be involved in the signaling process. PMID- 8228225 TI - A majority of Ig H chain cDNA of normal human adult blood lymphocytes resembles cDNA for fetal Ig and natural autoantibodies. AB - Certain Ig VH gene segments, with few or no mutations, recur frequently in natural autoantibodies, fetal antibodies, and products of B cell tumors. The goal of this study was to determine whether similar Ig gene segment usage occurs in normal human adult PBL. Extending previous analyses, 105 randomly picked H chain V region clones of representative cDNA libraries from PBL were sequenced. Clones were from: IgM and IgG libraries from one RNA sample of a normal adult; a second IgM library from the same subject 11 mo later; and one IgM library from a second subject. Although some clones had clear evidence of mutation, 48 of 77 IgM clones (62%) shared 99% or more identity with known germline VH segments, and most of these had no mutations in the CDR3 portion of the JH segment. Certain VH gene segments, expressed in autoantibodies and fetal antibodies, occurred at high frequency in these libraries. Fourteen of the clones with 99% identity to known VH segments had CDR3 segments identical to portions of known germline DH gene sequences; two such clones had no N nucleotides at the VHDH or DHJH junctions. IgG-encoding sequences had more mutations than IgM-encoding sequences. JH and DH usage was not random. The circulating B cell population may represent a distinct compartment, with a large proportion of cells similar to those of the fetal and natural autoantibody repertoire. Polyreactive Ig products of these circulating cells may serve a screening function, binding and delivering diverse Ag to secondary lymphoid tissues where more highly selective antibodies are formed to foreign or self-Ag. PMID- 8228226 TI - Characterization of platelet-activating factor receptors on canine T lymphocytes. AB - The ability of purified canine T lymphocytes to selectively bind platelet activating factor (PAF) was characterized. Authentic radiolabeled PAF rapidly and selectively bound to T lymphocytes and reached saturation within 1 min. This binding was reversible and highly selective for (R) PAF because (S) PAF, lyso PAF, and diacyl PAF did not displace the bound (R) PAF probe. Only increasing quantities of chemically pure (R) PAF displaced the radiolabeled (R) PAF probe. The binding maximum of PAF was determined to be 35 pM per 2 x 10(6) lymphocytes. Competitive radioligand binding studies and Scatchard analysis indicated a single class of high affinity receptors with a dissociation constant of 0.077 nM and a receptor density of 6419 receptors per cell. The ability of purified canine T lymphocytes to hydrolyze PAF to the biologically inactive metabolite lyso-PAF was also studied. Over a 30-min incubation period, about 5% of PAF was metabolized to lyso PAF. This rate of PAF hydrolysis was the same as the rate observed with the media without cells, suggesting a small degree of nonenzymatic hydrolysis. The effects of varying concentrations of authentic PAF on intracellular free Ca2+ release in purified T lymphocytes was evaluated using the fluorescent probe Fura 2 and excitation-emission spectrofluorometry. PAF below the concentration of 1.0 nM did not significantly increase intracellular Ca2+ in T lymphocytes. More than 1 nM PAF, intracellular-free Ca2+ modestly, but significantly, increased in T lymphocytes. In other experiments, canine PBMC proliferated in response to Con A and in the one way MLR. These proliferative responses were abolished when the selective PAF receptor antagonist SC-47014A was added to the culture medium. In the MLR, this inhibitory effect was dependent on the length of time that the antagonist was in the culture. Specifically, inhibition of proliferation was incrementally reversed when the PAF antagonist was introduced progressively later into the 7-day MLR stimulation period, suggesting that PAF receptor blockade prevents an MLR response from occurring, but is unable to suppress an existing MLR response. Although the Con A-induced mononuclear cell proliferation was abolished with PAF receptor antagonists, the addition of authentic biologically active PAF or PAF analogs did not alter the proliferative response to Con A. In conclusion, canine T lymphocytes possess high affinity receptors for PAF. These binding sites are highly selective and reversible. PAF binding slightly increases intracellular free Ca2+ in T lymphocytes and appears to be involved in lymphocyte proliferation in response to soluble plant mitogen and alloantigen. PMID- 8228227 TI - T cell receptor V alpha gene segment with alternate splicing in the junctional region. AB - The locus encoding mouse TCR-alpha chain includes approximately 100 V alpha gene segments that can be organized in about 20 structural subfamilies. Southern blot analysis of a T cell line derived from the BALB/c strain, M5T, has indicated that both alpha loci were rearranged, as assessed by the deletion of the delta locus, and that the V alpha gene segment involved in one of the rearrangements did not belong to any of the V alpha subfamilies already described. Transcripts of TCR alpha chains from the M5T line were cloned after cDNA synthesis and anchored polymerase chain reaction, revealing a V alpha gene segment of an as yet unidentified subfamily, V alpha 5T. Molecular cloning of germ-line V alpha 5T gene segments has shown that this subfamily contained two members, one of them being a pseudogene. The two members were located to each extremity of the alpha locus associated with a member of the V alpha 13 and V alpha BWB subfamilies. Analysis of transcripts bearing the V alpha 5T gene segment in the M5T line as well as in thymocytes has revealed that J alpha are frequently absent. This is due to an alternate donor splice site generated at the V alpha 5T-J alpha junction that leads to a splicing from the end of V alpha 5T to C alpha instead of the J alpha to C alpha conventional splicing. The impact of J alpha spliced out transcripts on the allelic exclusion process is discussed. PMID- 8228228 TI - Cloning and characterization of the 2B4 gene encoding a molecule associated with non-MHC-restricted killing mediated by activated natural killer cells and T cells. AB - We have recently described a signal transducing molecule, 2B4, expressed on all NK and T cells that mediate non-MHC-restricted killing. The gene encoding this molecule was cloned and its nucleotide sequence determined. The encoded protein of 398 amino acids has a leader peptide of 18 amino acids and a transmembrane region of 24 amino acids. The predicted protein has eight N-linked glycosylation sites, suggesting that it is highly glycosylated. Comparison of 2B4 with sequences in the databanks indicates that 2B4 is a member of Ig supergene family, and it shows homology to murine and rat CD48 and human LFA-3. Northern blot analysis has shown at least three transcripts for 2B4 in adherent lymphokine activated killer cells of several mouse strains and TCR-gamma/delta dendritic epidermal T cell lines but not in allospecific T cell clones. These three mRNA are the products of differential splicing of heterogeneous nuclear RNA. Southern blot analysis of genomic DNA from several mouse strains revealed that 2B4 belongs to a family of closely related genes. The 2B4 gene has been mapped to mouse chromosome 1 by analysis of 2B4 expression in recombinant inbred mouse strains. PMID- 8228229 TI - Polymorphism in the mouse Tap-1 gene. Association with abnormal CD8+ T cell development in the nonobese nondiabetic mouse. AB - Tap-1 and Tap-2 genes code for a heterodimeric peptide transporter required for the normal maturation and surface expression of class I molecules. Polymorphic variants of these MHC encoded genes occur in rats and humans. After failing to amplify a 3' polymerase chain reaction (PCR) product from thymic and splenic cDNA of the nonobese nondiabetic (NON) strain, we considered it possible that Tap-1 polymorphism was present, since cDNA from CBA/J, C57BL/6, BALB/c, and NOD (nonobese diabetic) mice all yielded Tap-1 3' products. Overlapping PCR fragments spanning the highly conserved ATP-binding cassette (ABC) were generated for purposes of restriction endonuclease analysis, studies of IFN-gamma regulation, and sequencing. To avoid amplifying other members of the transporter family, we used a gel-purified 1670-bp Tap-1 PCR "long product" as template for nested PCR. Sequencing revealed three polymorphic alleles. The most divergent was for the NON strain and involved two non-conserved amino acid substitutions (Arg-->Cys397 and Leu-->Arg491) and three silent mutations. NON mice show an abnormal pattern of class I (Kb) expression and a sizeable reduction in the percentage of CD8+ cells in the blood and thymus. In F2 segregants, the low CD8 phenotype mapped to the MHC. Tap-1 genes of NON and C57BL/6 mice were equally sensitive to up-regulation by IFN-gamma. We conclude that the mouse Tap-1 transporter gene, like the Tap-2 of the rat and the Tap-1 and Tap-2 of the human, is polymorphic. The extensive variation and specific codon changes of Tap-1 in the NON mouse raise the possibility that this gene is the MHC locus responsible for altering the intrathymic development of CD8+ T cells. PMID- 8228230 TI - Homogeneous antigen receptor beta-chain genes in cloned CD4- CD8- alpha beta T suppressor cells. AB - The rearrangements of beta-chain genes of the T cell Ag receptor were examined in 12 CD4- CD8- alpha beta + T cell lines derived from the spleen or thymus of neonatal or adult BALB/c mice. Eleven of the lines were cloned and established from six independent cloning procedures from different mice. Five cloned lines used V beta 9, four cloned lines used V beta 15, and two cloned lines used V beta 7. Nucleotide sequencing of the beta-chain genes showed that clones that used a given V beta were identically rearranged even when they were derived from independent cloning procedures. In the case of V beta 7 and V beta 15 all nucleotides in the V-D-J joining region were in the germ line configuration without N region additions. Rearrangements of the V beta 7, V beta 9, and V beta 15 genes were functional. Each V beta 15 clone also had a homogeneous rearrangement of the V beta 13 gene, which was nonfunctional. The predicted amino acid sequence of the joining regions of the V beta 7, V beta 9, and V beta 15 rearrangements showed homology in four of seven amino acids in the peptide contact region. PMID- 8228231 TI - The utilization of individual VH exons in the primary repertoire of adult BALB/c mice. AB - The utilization of VH gene families is severely biased during fetal development, such that D-region proximal VH genes are overrepresented. After birth the VH repertoire becomes more equally representative of the total inherited set of functional VH genes. To investigate the extent of this normalization process, we have determined the relative utilization of individual VH exons in the pre-immune repertoire of adult BALB/c spleen cells. Large samples of IgM heavy chain transcripts from polyclonally activated B cells were captured in a cDNA phage library and screened by hybridization using highly specific oligonucleotide probes. These studies revealed that the utilization of particular VH exons can differ by an order of magnitude. Significantly, the VH genes most under represented in the pre-immune repertoire are located in the region of the Igh locus most distal to the DjhCh region. We suggest that the chromosomal position of a particular VH gene may influence its utilization and that the normalization of the adult repertoire is incomplete. PMID- 8228232 TI - Isolation of a classical MHC class I cDNA from an amphibian. Evidence for only one class I locus in the Xenopus MHC. AB - The amphibian Xenopus is an ectothermic vertebrate in which the MHC has been studied extensively at the functional, biochemical, and genetic levels. A cDNA clone corresponding to the MHC class la gene (Xela-UAA1f) of Xenopus laevis was isolated by screening a cDNA phage library with oligonucleotides based on NH2 terminal protein sequence. Three pieces of evidence support its status as a class la gene: 1) Previous biochemical data suggested that only one polymorphic class la molecule is expressed per MHC haplotype in X. laevis. NH2-terminal sequencing of the class I protein encoded by the f haplotype showed a single unambiguous sequence of the first 22 amino acids; the deduced protein sequence of the cDNA clone matches precisely to this peptide sequence; 2) Genes that hybridized to the cDNA clone segregated perfectly with the serologically typed MHC in two family studies; and 3) There is a strong conservation of amino acids in the peptide binding region that have been shown in mammals to dock peptides at their NH2- and COOH-termini. In contrast to all other species that have been examined, there appears to be only one class I locus present in the MHC of X. laevis. Xenopus speciates by allopolyploidization, and there are Xenopus species with different levels of ploidy (2n-12n). Functionally, the MHC has been shown to be "diploidized" in most Xenopus species. As in previous studies with MHC class II and HSP70 probes, there is a trend toward maintaining a diploid number of class la genes in all Xenopus species regardless of their chromosome number, probably accomplished through a deletional mechanism. Thus, there is a strong pressure in Xenopus to maintain very few MHC-linked class I genes, exemplified both by the number of class I genes per MHC haplotype and by the number of class la genes per organism. PMID- 8228233 TI - Kinetics of E2A basic helix-loop-helix-protein expression during myelopoiesis and primary B cell differentiation. AB - The DNA-binding factors encoded by the E2A gene, which include the basic helix loop-helix proteins E12 and E47, are thought to regulate Ig gene expression. In the rat these factors are products of the Pan locus and are referred to as Pan-2 and Pan-1, respectively. To determine when during B cell differentiation Pan (E2A) DNA-binding proteins are first expressed, cells from a newly developed rat long term lymphoid bone marrow culture system were analyzed molecularly and phenotypically using a newly defined anti-Pan mAb. The data indicate that Pan expressing cells appear in B cell progenitors before the appearance of the CD45R B lineage-associated Ag or microH chain protein. The results also indicate that Pan proteins are present in cells that had undergone D-JH rearrangements. Finally, although Pan (E2A) proteins are present in myeloid precursors, expression is extinguished as differentiation into mature myeloid cells occurs. PMID- 8228234 TI - Effect of granulocyte/macrophage colony-stimulating factor on human monocytes infected with influenza A virus. Enhancement of virus replication, cytokine release, and cytotoxicity. AB - The activating properties of granulocyte/macrophage (GM)-CSF were studied in vitro with human monocytes infected by influenza A virus. When monocytes were pretreated for 8 h with GM-CSF (100 U/ml) and then exposed to influenza A virus, de novo virus protein synthesis was enhanced, more virus particles were released, and cells were killed at a higher rate. In virus-infected monocytes, GM-CSF induced a more rapid IFN-alpha release and potentiated production of TNF-alpha, IL-1 beta, and IL-6. Although GM-CSF or influenza A virus were each capable of independently activating TNF-alpha, IL-1 beta, and IL-6 gene transcription, a combination of both induced a massive cytokine mRNA accumulation which was readily translated into bioactive protein. Thus, GM-CSF may display a Janus-like action by accelerating virus infection but also by priming monocytes for elevated cytokine production. Whether the facilitated influenza A virus replication caused by GM-CSF may be counterbalanced by an improved cytokine response remains to be studied under more complex in vivo conditions. PMID- 8228235 TI - IL-10 neutralization augments mouse resistance to systemic Mycobacterium avium infections. AB - In this contribution, we examined the involvement of the cytokine IL-10 in the progression of experimental murine Mycobacterium avium infections in susceptible BALB/c mice. Addition of anti-IL-10 antibodies in the supernatants of peritoneal macrophages infected with virulent M. avium resulted in a significantly enhanced mycobacteriostatic activity of macrophages. In BALB/c mice infected with the B101 or B102 virulent M. avium strains, examination of the cytokine release profile in splenocytes from infected mice showed that infection was associated with an initial copious release of both IFN-gamma and IL-10. IL-10 production increased as the infection progressed, whereas IFN-gamma levels diminished. Infected mice were given repeated infusions of a rat mAb against mouse IL-10 or rat IgM. Examination of IgM serum levels in anti-IL-10-treated mice (infected or not) showed that depletion of endogenous IL-10 resulted in much decreased IgM levels. Results showed that infusions of large dosages of the monoclonal anti-IL-10 resulted in a very significantly diminished bacterial growth in the spleens. These findings indicate that IL-10 may have a negative impact on resistance to M. avium infections, due, at least in part, to decreased macrophage activity. PMID- 8228236 TI - Complement receptor type 3 (CD11b/CD18) involvement is essential for killing of Listeria monocytogenes by mouse macrophages. AB - Recent work indicated that C receptor type 3 (CR3) mediates most phagocytosis of the facultative intracellular bacterium Listeria monocytogenes by mouse macrophages, which can kill it. In contrast, phagocytosis of Listeria by a population of nonlistericidal macrophages was largely CR3-independent. These findings suggested that CR3 binding during phagocytosis may be important in determining whether a macrophage kills Listeria, or is parasitized by the bacterium. The experiments reported here tested this hypothesis. When phagocytosis and killing were assayed separately, normally listericidal peritoneal macrophages still could phagocytose to some extent, but lost listericidal activity when CR3 was blocked by mAb. Anti-CR3 mAb inhibited killing in a dose-dependent fashion, and at high doses the cells became permissive hosts. Microbicidal function also was inhibited when active C components were absent during phagocytosis and during killing. Because Listeria are confined to phagosomes in listericidal macrophages but escape into the cytoplasm in nonlistericidal macrophages, we tested whether anti-CR3 mAb enhanced phagosomal escape. In fact, escape of Listeria into the cytoplasm was rare in both control and anti-CR3 mAb-treated macrophages. Moreover, electron microscopy of these cells demonstrated dividing intraphagosomal bacteria. Taken together, these results suggest that binding to CR3 during phagocytosis leads to bacterial killing, and that phagocytic pathways engaged when binding to CR3 is blocked do not trigger microbicidal activity. Furthermore, restriction of Listeria to the phagosome in the absence of CR3 engagement is not by itself sufficient for macrophage listericidal activity. PMID- 8228237 TI - Role of acyl residues in polyclonal murine B cell activation by acylpoly(1,3)galactosides from Klebsiella pneumoniae. AB - Several components of Klebsiella pneumoniae including a membrane proteoglycan (Kp MPG) were reported to activate macrophages and to induce T-independent polyclonal activation of mouse B cells. Chemically defined derivatives of Kp-MPG were prepared and characterized, enabling us to approach the molecular substructures involved in the binding to lymphocytes and the activation of B cells. Five derivatives were characterized: (i) an acylpoly(1,3)galactoside containing ester linked fatty acids (EFA-APG) which was obtained by mild alkaline hydrolysis, (ii) a polymer of EFA-APG (APG pol1), (iii) a preparation obtained by drastic alkaline hydrolysis and delipidation which removed the esterified fatty acids (APG), (iv) a polymer of the latter compound (APG pol2), and (v) an APG preparation submitted to mild acid hydrolysis which removed all fatty acids but left the galactose chain of APG (GC-APG) intact. The derivatives were studied for their capacity to bind to and to activate mouse splenocytes. Binding was investigated on BALB/c and C3H/HeJ splenocytes by indirect immunofluorescence using biotinylated F(ab')2 of anti-Kp-MPG antibodies and the streptavidin-phycoerythrin amplification system in flow cytometry and by competition of unlabeled APG with biotinylated APG. Activation was studied by measuring (i) [3H]thymidine incorporation into spleen cells from BALB/c, C3H/HeJ, nude (nu+/nu+) mouse strains, and purified B cells of BALB/c; (ii) immunoglobulin secretion in culture supernatants; and (iii) blastogenesis. The results demonstrate a specific uptake of EFA-APG and APG by T cells as well as by B cells and exclude a contribution of the polygalactose part of the APG molecule (GC-APG) to the binding to spleen lymphocytes. Unlike LPS from the same strain of K. pneumoniae, APG pol1 stimulated B cell activation in the LPS-resistant C3H/HeJ strain as well as in BALB/c mice. The compounds did not activate T cells and were T-independent B cell activators, stimulating nu+/nu+ spleen cells and inducing primarily IgM and IgG3 synthesis. Polymers were more potent activators than monomers and removal of ester-linked fatty acids completely abrogated B cell-activating properties. The monomer APG antagonized B cell activation by Kp-MPG, LPS from K. pneumoniae, and APG pol1. The data indicate that within the EFA-APG molecule, distinct substructures are required for binding and for triggering B cell response. PMID- 8228238 TI - Tumor necrosis factor plays a role in Schistosoma mansoni egg-induced granulomatous inflammation. AB - In murine schistosomiasis mansoni several proinflammatory lymphokines participate in the circumoval granulomatous inflammatory response. At the acute stage of the infection lymphokine secretion is maximal and coincides with the strong granulomatous reaction. With chronicity, Ag-elicited lymphokine production diminishes and the granulomatous inflammation is downmodulated. In this study the role of TNF in the granulomatous response was investigated. Macrophages isolated from vigorous liver granulomas of acute infection mice on LPS stimulation produced significantly more TNF than their counterparts obtained from the downmodulated granulomas of chronic infection animals. Endotoxin-injected, acutely infected mice had higher TNF serum levels than those with chronic infection. Repeated injections of polyclonal anti-TNF-alpha antiserum given to acutely infected mice significantly suppressed the size of developing liver granulomas. Administration of graded dosages of recombinant murine TNF-alpha to mice with chronic infection restored the size of the downmodulated liver and lung granulomas to the level of the vigorous lesions. Repeated injections of rIL-2 to chronically infected mice augmented the downmodulated liver granulomatous response, induced enhanced production of TNF by the liver granuloma macrophages, and elevated cytokine levels in the sera of treated mice. Enhanced TNF production was also observed in vitro in monolayers of murine rIFN-gamma-treated granuloma macrophages. These findings indicate that focally and systemically produced TNF influences the development of the schistosome egg-induced granulomatous response. Moreover, in vivo manipulation of cytokine levels can modify the intensity of the inflammatory response. PMID- 8228239 TI - T cell receptor V beta 2 and V beta 6 mediate tumor-specific cytotoxicity by tumor-infiltrating lymphocytes in ovarian cancer. AB - The interaction between T lymphocytes and the Ag-HLA complex on tumor cells is mediated by the TCR. The diversity and the specificity of the TCR are in part secondary to the gene rearrangement of the V region on the beta-chain (V beta). To determine whether a restricted number of TCR V beta genes are utilized in the recognition of ovarian cancer, tumor-infiltrating lymphocytes (TIL) were isolated from six consecutive untreated ovarian cancer patients. TIL were also cultured using repeated autologous tumor stimulation, and by 7 wk, five of six patients produced bulk cultures consisting of > 50% CD8+ T cells and demonstrating an autologous tumor-specific pattern of cytotoxicity. TCR V beta gene usage was analyzed in the five patients yielding fresh TIL and corresponding 7-wk cultured, tumor-specific TIL; 22 primers specific for 20 TCR V beta gene families were employed and amplified by polymerase chain reaction and then quantitated by HPLC. A heterogeneous pattern of V beta usage was seen in the fresh TIL; however, V beta 2, V beta 3, V beta 6, V beta 7, V beta 8, and V beta 13.1 were found in increased proportions in at least three of five patients. In the 7-wk tumor specific TIL, V beta analysis showed an increased usage of V beta 2, V beta 3, V beta 6, and V beta 7 in more than three of five patients. No significant change in V beta representation was seen in control populations that were not stimulated with tumor. Looking at the percent change in V beta usage between fresh and 7-wk tumor-specific cultures, V beta 2 and V beta 6 were augmented significantly in at least three of five patients (108% and 61%, respectively). To verify that the increase in representation of these V beta families was responsible for the increased cytotoxicity observed, mAb specific for V beta 2 and V beta 6 were used to block tumor lysis. Anti-V beta 6 and anti-V beta 2 significantly blocked cytotoxicity against autologous tumor cells in those TIL populations expressing increased levels of these V beta families. These data suggest that a selective repertoire of TCR V beta genes is used to recognize the Ag-HLA class 1 complexes on the surface of ovarian tumor cells, and specifically V beta 2 and V beta 6 appear to mediate antitumor activity. These findings may aid in the development of a more specific immunotherapy in ovarian cancer. PMID- 8228240 TI - HLA-A2 presents shared tumor-associated antigens derived from endogenous proteins in ovarian cancer. AB - Tumor-associated lymphocytes (TAL) from the malignant ascites and tumor infiltrating lymphocytes (TIL) from the solid tumor were isolated from six consecutive untreated ovarian cancer patients. Tumor-specific CTL were generated from both TAL and TIL using solid phase anti-CD3, low dose IL-2 (50 IU/ml), and repeated tumor stimulation. The specificity of TAL and TIL was tested in standard cytotoxicity assays using autologous tumor, several allogeneic ovarian tumors, and the NK-sensitive cell line, K562. Anti-HLA-A-B-C mAb, W6/32, was used to demonstrate that these tumor-specific TAL and TIL were HLA class I-restricted. The ability of the ascitic and solid tumor to present Ag by HLA class I was assessed using Brefeldin A, a fungal metabolite that blocks the endogenous Ag processing pathway in the viral model. Brefeldin A significantly inhibited tumor specific cytotoxicity as well as HLA class I expression on the cell surface, suggesting an endogenous source of tumor-associated Ag. Despite previous reports of antigenic heterogeneity in ovarian cancer, shared tumor-associated Ag were shown to exist in this disease as demonstrated by significant allogeneic recognition of HLA-A2-matched patients as opposed to unmatched controls. Specifically, CTL from HLA-A2+ patients lysed HLA-A2+ allogeneic targets significantly better than HLA-A2- allogeneic or HLA-A2+ melanoma targets. There was no such difference with HLA-A2- effectors. Furthermore, HLA-A2 was confirmed to be a major restriction element in ovarian cancer by the blocking of HLA-A2+ effectors against both autologous and allogeneic HLA-A2+ targets with the anti HLA-A2 mAb, BB7.2. These findings verify a similar lymphocyte/tumor interaction as has been documented in melanoma, suggesting a common mechanism of recognition of these human tumors by lymphocytes. PMID- 8228241 TI - Suppressor macrophages in African trypanosomiasis inhibit T cell proliferative responses by nitric oxide and prostaglandins. AB - Suppression of host T cell responses is one of the hallmarks of infection with the African trypanosomes. The cellular basis for immunosuppression includes the generation of suppressor macrophages that down-regulate T cell proliferative but not necessarily cytokine responses to both mitogen and trypanosome Ag. Since macrophages from infected animals display activation characteristics, we have asked whether products of activated cells, specifically nitric oxide (NO) and PG, may mediate the suppressor cell effects and immunosuppression observed. We demonstrate that cells isolated from B10.BR mice infected with Trypanosoma brucei rhodesiense exhibited transcriptional up-regulation of inducible NO synthase and released significant amounts of NO. The levels of NO released were elevated further after stimulation of cells with T cell mitogens or specific parasite Ag; antibody blocking experiments demonstrated that this up-regulation of NO synthesis was at least partially dependent upon IFN-gamma and TNF-alpha. The addition of inducible NO synthase substrate analogues such as NG-monomethyl-L arginine to cell cultures inhibited NO release and also partially reversed the suppressor cell activity and immunosuppression displayed by such cultures. PG levels also were elevated in cell cultures from infected mice, but the PG inhibitor indomethacin had no effect on suppressor cells or suppression when added alone to the cultures. However, the concurrent inhibition of NO and PG synthesis by the addition of both NG-monomethyl-L-arginine and indomethacin completely blocked suppressor cell activity associated with infected macrophages and also resulted in further recovery of infected cells from immunosuppression, thus revealing an epistatic effect between these two mediators. We conclude that macrophage activation in trypanosomiasis induces the release of reactive nitrogen intermediates and PG, which down-regulate proliferative responses by T cells during infection. PMID- 8228242 TI - 70 kDa heat shock cognate protein is a transformation-associated antigen and a possible target for the host's anti-tumor immunity. AB - We previously investigated a novel heat-inducible transformation-associated cell surface Ag that is expressed on the activated H-ras oncogene-transformed rat fibrosarcoma W31, but not its parental nontransformed fibroblast WFB. This Ag was detected by mAb 067. Herein, we characterized the molecular nature of the Ag by using anti-heat shock protein (HSP) mAb. The accumulated data indicated that the cell surface expression of Ag was clearly enhanced by several stressors, such as TNF, L-azetidine-2-carboxylic acid, and sodium arsenite. The immunoprecipitate made with mAb 067 and W31 cell lysates reacted with anti-rat 70 kDa heat shock cognate (HSC) mAb, TG5E, indicating that 067-defined Ag may be a rat 70 kDa HSC. Because this Ag seemed to be one of the transformation-associated Ag of WFB, we further studied whether it could play an important role in the host's anti-tumor immunity. Peripheral T cells of rats primed with live BCG showed cytotoxicity to W31 but not to WFB. Because the possibility existed that HSP may interact with certain populations of T cells, we focused on the reactivity of CD4-CD8- double negative T (DNT) cells against 067-defined molecule. DNT cells from spleen and PBL of live BCG-primed rats showed the cytotoxicity against W31 cells. This cytotoxicity was completely blocked by mAb 067 and anti-CD3 mAb. However, it was not blocked by mAb R48B1 and 109, which detect the MHC class I nonpolymorphic determinant and a target molecule of the cytolysis by poly I:C-induced NK cells, respectively. Furthermore, brefeldin A was able to block the cytotoxicity against W31 targets by DNT cells, but not by NK cells. These data suggest that 70 kDa HSC may be a tumor Ag and may act as a presenting molecule perhaps complexed with cellular peptides to certain DNT cells. PMID- 8228243 TI - Mechanisms of the inflammatory response induced by extracts of Schistosoma mansoni larvae in guinea pig skin. AB - Penetration of skin by cercariae of Schistosoma mansoni is associated with a local inflammatory response characterised by leukocyte accumulation and tissue swelling. The significance of the inflammation and its relevance to infection of the host is unknown. In this study, we have investigated the mechanisms of the local inflammatory response induced by injecting extracts of Schistosoma mansoni cercariae into guinea pig skin. Intradermal injection of cercarial homogenate or cercarial transformation fluid induced a dose-dependent increase in local edema formation and accumulation of intravenously injected 111In-labeled eosinophils and neutrophils. The responses were rapid in onset and independent of new protein synthesis. The capacity of extracts to induce edema formation and leukocyte accumulation correlated significantly with their proteolytic activity. In addition, extract-induced inflammation was reduced by co-injection with Trasylol, soybean trypsin inhibitor, PMSF, or heparin. A combination of PMSF and heparin virtually abolished extract-induced inflammation. The known inhibitory effect of Trasylol on kallikrein prompted an investigation into the role of kinins in inducing local edema formation. The bradykinin antagonist HOE 140 substantially reduced extract-induced edema whereas bradykinin itself was weak at inducing leukocyte accumulation. The cyclooxygenase inhibitor ibuprofen, the platelet activating factor antagonist WEB 2086, and the 5-lipoxygenase inhibitor PF5901 had no effect on edema formation. In contrast, extract-induced eosinophil accumulation was reduced by WEB 2086 and PF5901, suggesting that endogenous PAF and leukotriene B4 were involved in recruiting these cells to inflammatory sites. Thus, cercarial proteases induce a local inflammatory response in guinea pig skin that can be attributed, in part, to known mediators of inflammation. It remains to be established whether the response is protective for the host or whether it is beneficial for the parasite. Further understanding of the underlying mechanisms may provide a target for the pharmacologic control of infection in schistosomiasis. PMID- 8228244 TI - Requirement of thiol compounds as reducing agents for IL-2-mediated induction of LAK activity and proliferation of human NK cells. AB - Thiol-related compounds, such as L-cystine, 2-ME or reduced glutathione (GSH), are important in many lymphoid cell activation pathways. We investigated their role in IL-2-generated lymphokine-activated killer (LAK) activity in human NK (CD16+ and/or CD56+) cells. Depletion of GSH and L-cystine, but not L-leucine or glycine, from medium used during culture of NK cells with IL-2 inhibited LAK and proliferative activities, whereas IL-2-independent lytic function of NK cells remained intact. Addition of L-cysteine, 2-ME or GSH, but not methylated analogs of these compounds (which cannot function as proton donors) to L-cystine/GSH depleted medium restored proliferative response of NK cells and LAK generation. In the presence of L-buthionine-(S,R)-sulfoximine, an inhibitor of GSH synthesis, IL-2-induced LAK activity and proliferation of NK cells in medium without L cystine and GSH, could be restored, at least in part, by addition of GSH, but not 2-ME or L-cystine. Furthermore, intracellular GSH levels were depressed in cells cultured in L-cystine/GSH-depleted medium but could be restored by the addition of 2-ME. The results suggest that (1) L-cystine or thiol containing compounds such as L-cysteine, 2-ME, or GSH are necessary for effective IL-2-activation of human NK cells, (2) these compounds must be functional proton-donors, i.e., reducing agents, implying regulation of the IL-2 activation pathway by oxidation reduction, and (3) GSH synthesis is necessary for the activation. Experiments were performed to begin to dissect the redox-sensitive step(s) in LAK development. Depletion of reducing agents had no effect on internalization of rIL 2. In contrast, intracellular granzyme A activity was significantly depressed in NK cells cultured with rIL-2 in L-cystine/GSH-depleted medium compared with those cultured in medium in which L-cystine levels had been replenished. The findings suggest that step(s) in the transduction of the IL-2 signal in NK cells, between the internalization of IL-2 and the maturation of the lytic mechanism, are subject to regulation by oxidation-reduction. PMID- 8228245 TI - Characterization of factor H binding to human polymorphonuclear leukocytes. AB - Previous studies indicate that factor H (fH) binds to a number of cell types and may have functions other than C regulation. We have examined for fH binding to PMN using flow cytometry and radiolabeled binding assays. Binding of fH was demonstrated to be specific and saturable with approximately 6 x 10(4) binding sites/polymorphonuclear leukocytes (PMN) and a Ka value of 3.3 x 10(8) L/M. Binding of fH to PMN was ionic strength dependent, required divalent cations, and was enhanced by PMN stimulation with FMLP and calcium ionophore, A23187. The 38 kDa N-terminal tryptic fragment of fH bound to PMN and blocking experiments with mAb suggested a receptor binding site was located within the fifth SCR of fH. fH binding was not due to associations with surface-bound C3 or to CR3. Binding of fH to U937 and Raji cells, but not to T cells was also demonstrated. These studies provide presumptive evidence for a fHR on PMN. Binding of fH by fHR could enhance recognition of opsonized targets, trigger secondary intracellular events or contribute to intrinsic protection of cells against C. PMID- 8228246 TI - LPS-induced sTNF-receptor release in vivo in a murine model. Investigation of the role of tumor necrosis factor, IL-1, leukemia inhibiting factor, and IFN-gamma. AB - TNF, a primary mediator of the response to infection, can be injurious to the organism when present in excessive quantities. Circulating soluble TNF receptors (sTNFR) appear to represent a natural mechanism that protects against circulating TNF. Two soluble TNF receptors (sTNFR-P55 and sTNFR-P75) circulate in vivo and are up-regulated in response to endotoxin. In this study, we investigated the kinetics of LPS-induced sTNFR release and the role of the cytokines TNF, leukemia inhibiting factor, IFN-gamma, and IL-1 in this process. The results show that LPS injection results in a rapid increase in levels of both sTNFR. Although sTNFR-P55 decreases after a peak at 30 min, sTNFR-P75 levels show a peak after 4 to 8 h, after which they slowly diminish. Both human TNF and murine TNF are capable of increasing levels of both sTNFR. Blocking circulating TNF by administration of 3 different anti-TNF agents before LPS injection (mAb to murine TNF, sTNFR55-Fc or sTNFR75-Fc) results in a significant increase of sTNFR-P55 levels, whereas only both sTNFR-Fc constructs also significantly increase sTNFR-P75 levels. Although IL-1 receptor antagonist pretreatment before LPS has no effect on TNF or sTNFR levels, leukemia inhibiting factor pretreatment significantly increases sTNFR-P55 levels. Pretreatment with anti IFN-gamma mAb before LPS results in a significant reduction in TNF and sTNFR-P55 levels, but sTNFR-P75 levels are significantly increased. Our data show that both sTNFR can be up-regulated by LPS and TNF. The influence of TNF, leukemia inhibiting factor, IL-1, and IFN-gamma on the kinetics of LPS-induced circulating sTNFR is discussed in the context of the pathophysiology of LPS-induced disease. PMID- 8228247 TI - Dissociation of IL-1 beta synthesis and secretion in human blood monocytes stimulated with bacterial cell wall products. AB - Human blood monocytes synthesize but do not secrete IL-1 beta in response to low doses of bacterial cell-wall products. With this observation, we have developed a two-step staged release assay that separates IL-1 beta synthesis from secretion. This assay can be used to identify secretagogues or inhibitors of IL-1 beta secretion as well as the biochemical events leading to IL-1 beta release. Human blood monocytes are first treated with low (< 50 pg/ml) doses of LPS, which causes the synthesis of intracellular proIL-1 beta. Release of intracellular IL-1 beta can be induced by further treatment with 100 ng/ml of LPS or 1 x 10(6) CFU/ml of heat-killed Staphylococcus aureus. The amount and the efficiency of IL 1 beta secretion in the staged release assay was comparable with that of a standard method of treating blood monocytes with a single dose of 100 ng/ml LPS. Ongoing protein synthesis was not required for IL-1 beta secretion because mature IL-1 beta release occurred in the presence of the protein synthesis inhibitor cycloheximide. We have compared the effects of four different inhibitors of cytokine synthesis on IL-1 beta production in the standard and staged release assays. We find that dexamethasone or IL-10, when added together with 100 ng/ml LPS, inhibits IL-1 beta production with IC50 levels of 0.2 microM and 2.0 ng/ml, respectively. The IC50 levels increase greater than 50-fold when tested against monocytes pretreated with 50 pg/ml of LPS. These data suggest that dexamethasone and IL-10 have little effect on IL-1 beta secretion. Conversely, the IL-1 beta converting enzyme inhibitor, Ac-Tyr-Val-Ala-Asp-CHO (L-709,049) and the anti inflammatory agent [5-(4-pyridyl)6(4-fluorophenyl)-2,3-dihydroimidazo(2,1 b)thiazole] (SK&F 86002) inhibited IL-1 beta release in both the standard and staged release assays with IC50 of 1 microM. The data suggest that L-709,049 and SK&F 86002 interfere with steps involved in IL-1 beta secretion, as opposed to synthesis. The results also document a novel method for delineating separate events in the pathway required for IL-1 beta biosynthesis and further distinguish two classes of compounds capable of modulating IL-1 beta secretion. PMID- 8228248 TI - Antigen and carbachol mobilize calcium by similar mechanisms in a transfected mast cell line (RBL-2H3 cells) that expresses ml muscarinic receptors. AB - Because of unresolved questions about the mechanism of Ag-stimulated Ca2+ influx, Ca2+ mobilization in response to carbachol and Ag was compared in transfected rat basophilic RBL-2H3(ml) cells that expressed both Fc epsilon and ml muscarinic receptors. Although the stimulants activated phospholipase C via different coupling mechanisms, a G protein for carbachol or a tyrosine kinase for Ag, they released Ca2+ from the same intracellular pool and used the same or very similar mechanisms for influx of Ca2+ as indicated by the similar patterns of inhibition of uptake of 45Ca2+ by various cations. With both stimulants, influx and sustained increases in free cytosolic Ca2+ ([Ca2+]i) were associated with relatively small increases in inositol 1,4,5-trisphosphate (IP3). Blockade of Ca2+ influx resulted in rapid decline in [Ca2+]i to basal levels; resumption of influx caused a substantial "spike" in [Ca2+]i before [Ca2+]i reequilibrated at the same former steady-state levels but without perturbing levels of IP3. Thus, the refilling and discharge of Ca2+ from IP3-sensitive stores might occur synchronously on resumption of influx, or asynchronously during sustained influx and elevation of [Ca2+]i. Together, the results suggested that influx of Ca2+ in response to stimulation via Fc epsilon receptors occurred through a pathway, analogous to that observed with other types of stimulants, in which Ca2+ influx follows emptying of intracellular Ca2+ stores by IP3. Also, secretion was highly dependent on this IP3-dependent pathway. PMID- 8228249 TI - Tissue-specific regulation of IL-6 production by IL-4. Differential effects of IL 4 on nuclear factor-kappa B activity in monocytes and fibroblasts. AB - IL-4 inhibits production of certain proinflammatory cytokines, including IL-1 beta, TNF-alpha, and IL-6, by activated monocytes. Although monocytes are a major source of IL-6, other cell types such as fibroblasts and endothelial cells can also express this cytokine. To determine whether IL-4 inhibits IL-6 expression in non-hemopoietic cells, we investigated the effects of IL-4 on IL-6 production in both primary human fibroblasts and fibroblast lines. Rheumatoid synovial fibroblasts were evaluated in these studies because, like monocytes, they produce high levels of IL-6 when stimulated with IL-1. Although peripheral blood monocytes did not constitutively express IL-6 mRNA or protein, stimulation with IL-1 or LPS induced de novo IL-6 expression in these cells. In contrast, synovial fibroblasts displayed a significant basal level of IL-6 production, which was markedly increased after stimulation with IL-1. IL-4 suppressed IL-6 expression in monocytes, but did not inhibit IL-6 production in synovial fibroblasts. The inability of IL-4 to suppress IL-6 synthesis in rheumatoid synovial fibroblasts was not caused by a lack of IL-4R and was not unique to these cells because IL-4 also failed to inhibit IL-6 production in normal fibroblast lines derived from other tissues. Inhibition of IL-6 production by IL-4 in monocytes was associated with decreased nuclear NF-kappa B levels. However, IL-4 does not globally suppress the activity of all DNA-binding proteins because IL-4 treatment did not reduce the levels of NF-IL-6 or NF-IL-1 beta B in the same cells. Because NF kappa B activation is required for transcription of many cytokine genes, including IL-6, the ability of IL-4 to suppress NF-kappa B activity in monocytes suggests a potential mechanism by which this molecule may inhibit the expression of multiple cytokines. PMID- 8228250 TI - Increased synthesis and secretion of a 14-kDa phospholipase A2 by guinea pig alveolar macrophages. Dissociation from arachidonic acid liberation and modulation by dexamethasone. AB - The occurrence of a 14-kDa secretory phospholipase A2 (PLA2) in guinea pig alveolar macrophages (AM) and its relationship with the release of arachidonic acid (AA) were investigated. Freshly collected AM showed no detectable PLA2 activity as measured by the in vitro hydrolysis of phosphatidic acid. However, the PLA2 activity increased progressively when AM were maintained in culture to reach a level 60- to 100-fold greater than basal values within 20 h, with a parallel secretion into the incubation medium. By contrast, the activities of other phospholipid-hydrolyzing enzymes (platelet-activating factor acetylhydrolase and lysophospholipase) were modified only marginally. Both intra- and extracellular increases of PLA2 activity were abrogated with actinomycin D or cycloheximide. The enhanced PLA2 activity preferentially hydrolyzed negatively charged phospholipids in the order phosphatidic acid > phosphatidylglycerol > phosphatidylethanolamine > phosphatidylcholine, had an optimum pH of 7.5, and required a millimolar Ca2+ concentration for optimal activity and an apparent molecular mass of 14 kDa. Taken together, these results suggest that cultured AM elaborate an enzyme similar to the group II PLA2. On the other hand, our results show that AM hydrolyzed exogenous 2-arachidonoyl phosphatidylcholine and released AA and metabolites on FMLP stimulation. However, in contrast to the increase observed in the activity of the 14-kDa PLA2, the enzymatic activity involved in the hydrolysis of 2-arachidonoyl phosphatidylcholine and AA release remained constant with the culture duration of AM. Finally, dexamethasone markedly inhibited the increase of PLA2 activity, but only marginally inhibited the release of AA and metabolites from FMLP-stimulated AM. We conclude that guinea pig AM elaborate a 14-kDa PLA2 similar to the group II PLA2 through RNA- and protein synthesis-dependent processes. This elaboration appears to be induced by the adhesion of AM and is clearly dissociated from the liberation of AA. PMID- 8228251 TI - Regulation of eosinophil migration by adult T cell leukemia-derived factor. AB - Adult T cell leukemia-derived factor (ADF), originally defined as an IL-2 receptor alpha-chain (IL-2R alpha)/p55 (Tac) inducer, is a human thioredoxin homologue and has many cytokine-like activities. In this study, we examined the regulatory effect of ADF on eosinophil migration using human eosinophils and an eosinophilic subline of HL-60 human promyelocytic leukemia cells, YY-1. rADF induced migration of eosinophils from patients with hypereosinophilia, although rADF exhibited little activity on eosinophils from healthy donors. When human eosinophils were incubated with rADF (0.1-10 micrograms/ml) at 37 degrees C for 24 h, both chemotactic and chemokinetic activity of the complement anaphylatoxin peptide C5a on eosinophil migration was markedly enhanced in a dose-dependent manner. Similarly, this enhancing effect of rADF was observed in the migration assay using YY-1 cells. In contrast, rADF showed no modulation of migratory behavior of human eosinophils and YY-1 cells by IL-3, IL-5, nor granulocyte macrophage colony-stimulating factor. Scatchard analysis of C5a receptors on YY-1 cells using 125I-C5a showed that rADF modulated neither the density nor the affinity of the cell membrane significantly. Furthermore, mutant ADF (mADF), which had no reducing activity, had no enhancing effect on C5a-induced eosinophil migration. These results indicate a possible involvement of ADF in the recruitment of eosinophils through redox regulation by a dithiol reductase activity. PMID- 8228252 TI - Mechanism of release of soluble forms of tumor necrosis factor/lymphotoxin receptors by phorbol myristate acetate-stimulated human THP-1 cells in vitro. AB - The mechanism involved in the release of the soluble forms of 55 and 75 kDa TNF and lymphotoxin (LT) membrane receptors was studied in a continuous human monocytic cell line, THP-1, in vitro. THP-1 cells were found to spontaneously release soluble forms of both 55 and 75 kDa TNF/LT receptors. Release was up regulated by PMA, and optimal release was achieved at 10(-8) M PMA. Serine protease inhibitors such as PMSF,3,4 dichloroisocoumarin, N alpha-p-tosyl-L lysine chloromethyl ketone (TLCK), and N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) were found to inhibit the production of both soluble TNF/LT receptors. PMSF (2 mM) also blocked receptors shedding from paraformaldehyde fixed THP-1 cells coincubated with conditioned media from PMA-stimulated THP-1 cells. Colchicine at 1 and 10 microM stimulated the production of both soluble TNF/LT receptors, but the PMA-induced release of both soluble TNF/LT receptors was inhibited. It appears that the PMA-induced release of soluble TNF/LT receptors involves serine proteases in the extracellular space where the soluble parts of the TNF/LT receptors are cleaved directly off the cell membrane. PMID- 8228253 TI - Regulation of CD11b/CD18 expression in human neutrophils by phospholipase A2. AB - Recent evidence suggests that phospholipase A2 (PLA2)-derived lipid mediators may regulate a number of neutrophil responses including degranulation and adhesion. In view of the potential role of PLA2 in stimulus-secretion coupling, we examined the relationship between PLA2 activation and the surface expression of CD11b/CD18 (MAC-1) in human polymorphonuclear leukocytes (hPMNL), including the functional consequences of PLA2 inactivation on MAC-1-dependent adhesion. The selective inhibition of PLA2 by the marine natural products manoalide (MLD) and scalaradial (SLD) blocks [3H]arachidonic acid (AA) release in calcium ionophore A23187 stimulated neutrophils, and also inhibits secretion of specific and azurophilic granule constituents. Additional studies demonstrate that MLD, SLD, and other less potent PLA2 inhibitors such as 4-bromophenacylbromide and nordihydroguiaretic acid inhibit the surface expression of MAC-1 (IC50: MLD, 0.33 microM; SLD, 0.23 microM; 4-bromophenacylbromide, 2.8 microM; NDGA, 3.5 microM) at concentrations similar to those at which they inhibit [3H]AA release. Inhibitors of cyclooxygenase, 5-lipoxygenase, protein kinase C, or calcium channel antagonists have no effect on MAC-1 expression. PLA2 inactivation also prevents MAC-1 up-regulation in hPMNL stimulated with FMLP, IL-8, TNF-alpha, PMA, or platelet activating factor. In FMLP-stimulated hPMNL, under conditions in which no secondary granule constituents are secreted, MAC-1 and alkaline phosphatase up-regulation from intracellular granules is inhibited by MLD and SLD. Functional assays also demonstrate that MLD and SLD block MAC-1-dependent adhesion of activated neutrophils to keyhole limpet hemocyanin at concentrations that block the surface expression of MAC-1. [3H]AA release and MAC-1 expression in MLD and SLD-treated hPMNL could be recovered in the presence of 1 mM hydroxylamine in a time-dependent fashion, consistent with reported data that MLD and SLD inactivate PLA2 through Schiff base formation. In summary, these data emphasize the role of PLA2 as a key regulator of MAC-1 expression in models of neutrophil adhesion. PMID- 8228254 TI - Functional molecular complexes of human N-formyl chemoattractant receptors and actin. AB - When human neutrophils become desensitized to formyl peptide chemoattractants, the receptors (FPR) for these peptides are converted to a high affinity, GTP insensitive form that is associated with the Triton X-100-insoluble membrane skeleton from surface membrane domains. These domains are actin and fodrin-rich, but G protein-depleted suggesting that FPR shuttling between G protein-enriched and depleted domains may control signal transduction. To determine the molecular basis for FPR interaction with the membrane skeleton, neutrophil subcellular fractions were screened for molecules that could bind photoaffinity radioiodinated FPR solubilized in Triton X-100. These receptors showed a propensity to bind to a 41- to 43-kDa protein band on nitrocellulose overlays of SDS-PAGE-separated cytosol and plasma membrane fractions of neutrophils. This binding, as well as FPR binding to purified neutrophil actin, was inhibited 50% by 0.6 microM free neutrophil cytosolic actin. Addition of greater than 1 microM G-actin to crude or lectin-purified Triton X-100 extracts of FPR from neutrophil membranes increased the sedimentation rate of a significant fraction of FPR two to three fold as measured by velocity sedimentation in Triton X-100-containing linear sucrose density gradients. Addition of anti-actin antibodies to FPR extracts caused a concentration-dependent immunoprecipitation of at least 65% of the FPR. More than 40% of the immunoprecipitated FPR was specifically retained on protein A affinity matrices. Membrane actin was stabilized to alkaline washing when membranes were photoaffinity labeled. Conversely, when purified neutrophil cytosolic actin was added to membranes or their digitonin extracts, after prior depletion of actin by an alkaline membrane wash, photoaffinity labeling of FPR was increased two- to fourfold with an EC50 of approximately 0.1 microM actin. We conclude that FPR from human neutrophils may interact with actin in membranes to form Triton X-100-stable physical complexes. These complexes can accept additional G-actin monomers to form higher order molecular complexes. Formation of FPR-actin complexes in the neutrophil may play a role in the regulation of chemoattractant-induced activation or actin polymerization. PMID- 8228255 TI - Triggering of degranulation in mast cells by exogenous type II phospholipase A2. AB - We have previously shown the possibility that endogenous type II phospholipase A2 (PLA2) might participate in degranulation in mast cells (MC) (Murakami, M., et al. 1992. Eur. J. Biochem. 209:257). Now we have examined whether or not exogenously added type II PLA2 triggers MC degranulation. When rat peritoneal connective tissue MC (CTMC) were exposed to purified rat type II PLA2 at concentrations of more than 10 micrograms/ml, significant release of histamine was observed, whereas PGD2 was not generated under the same conditions. Mouse peritoneal CTMC as well as bone marrow-derived immature MC also responded to PLA2. Preincubation of CTMC with tyrosine kinase inhibitors, genistein, and herbimycin A, but not with pertussis toxin, resulted in abolition of the sensitivity to PLA2. The ability of type II PLA2 to induce histamine release was inhibited by an antibody or chemicals, both of which blocked the catalytic activity of type II PLA2. Heparin or an antibody recognizing the heparin-binding domain of type II PLA2 also suppressed the MC-degranulating activity, probably due to inhibition of binding of PLA2 to the cells. The interaction between heparan sulfate on cell surface and the heparin-binding domain of type II PLA2 may be important for the induction of exocytosis. The catalytic domain of the enzyme is also crucially important for the degranulation induction. Furthermore, we found that nerve growth factor, one of the potent regulators of MC function, significantly potentiated type II PLA2-induced histamine release from rat CTMC. These results suggest the possible role of extracellular type II PLA2 in activation of CTMC primed with nerve growth factor at inflamed sites. PMID- 8228256 TI - Signal transducing properties of the N-formyl peptide receptor expressed in undifferentiated HL60 cells. AB - Differentiated HL60 cells respond to challenge with ligand by mobilizing intracellular second messengers, resulting in superoxide production, degranulation, and actin polymerization with subsequent chemotaxis and phagocytosis. The functional capabilities of undifferentiated HL60 cells have not been similarly characterized due to the absence of the cell surface receptors required to initiate these processes. To investigate these properties, undifferentiated HL60 cells were transfected with one of the better characterized neutrophil chemotactic receptors, the N-formyl peptide receptor (FPR). Expression of the recombinant FPR gene product in FPR-transfected HL60 cells and the absence of the endogenous FPR in vector-transfected HL60 cells was demonstrated by Northern blot and flow cytometric analyses. FPR-transfected HL60 cells retained their ability to undergo granulocytic differentiation with dibutyryl cAMP, as determined by FMLP- and PMA-stimulated superoxide production. Furthermore, incubation of FPR-transfected HL60 cells for 5 days in the presence of FMLP resulted in limited differentiation as evidenced by the expression of functional C5a receptors. Binding studies of FPR-transfected HL60 cells demonstrated the presence of two binding affinities with dissociation constants of 0.6 and 33 nM, similar to dibutyryl cAMP differentiated HL60 cells and human neutrophils but contrasting the single high affinity state of the FPR expressed in mouse L cell fibroblasts. FPR-transfected HL60 cells displayed FMLP-dependent calcium mobilization with an EC50 of 3 nM and actin polymerization with an EC50 of approximately 10 nM. Actin polymerization was not observed in FPR-transfected L cell fibroblasts or undifferentiated vector-transfected HL60 cells. Both calcium mobilization and actin polymerization were sensitive to treatment with pertussis toxin, indicating the requirement for a Gi-like protein. Stimulation of either undifferentiated or differentiated HL60 cells with ATP resulted in pertussis toxin-insensitive calcium mobilization but was ineffective in producing actin polymerization. The results described herein show for the first time that undifferentiated HL60 cells can respond to chemoattractant receptor stimulation with many of the properties of the mature neutrophil. Transfected HL60 cells will provide an excellent system to study the characteristics of chemotactic receptors as well as the functional properties of myeloid cells. PMID- 8228257 TI - Activated eosinophils evoke chloride secretion in model intestinal epithelia primarily via regulated release of 5'-AMP. AB - Eosinophils may be prominent in intestinal diseases including allergic gastroenteritis, inflammatory bowel disease, enteritis associated with hypereosinophilic syndromes (HES), and parasitic diseases. Unlike normal blood eosinophils, those that circulate in HES and those that infiltrate inflamed tissue exhibit an "activated" phenotype. To model intestinal epithelial eosinophil interactions, we used peripheral blood eosinophils and human crypt like T84 epithelial cell-line monolayers. Eosinophils from normal, mildly atopic donors, only if activated by PMA or primed with granulocyte-macrophage-CSF for 48 h, as well as eosinophils from HES patients elicited a short circuit current when applied apically to T84 monolayers. This eosinophil-derived bioactivity, which was transferable in cell-free supernatants and in < 1000 m.w. ultrafiltrates, stimulated electrogenic Cl- secretion, as indicated by inhibition with basolateral bumetanide or gluconate substitution and by enhancement of the rate constant for 125I efflux from preloaded T84 cells. This secretagogue activity was blocked in both intact activated eosinophils and in eosinophil-conditioned supernatants, by 8-phenyl-theophylline, indicating involvement of an adenosine receptor. Ion exchange and reversed-phase HPLC analyses demonstrated that eosinophil supernatant ultrafiltrates contained elevated levels of 5'-AMP that was converted to adenosine after incubation with epithelium. Inhibition of epithelial apical membrane ecto-5'-nucleotidase ablated the conversion to adenosine. These studies establish that activated eosinophils elicit Cl- secretion from intestinal epithelial and that 5'-AMP released by eosinophils followed by its conversion to adenosine at the epithelial surface is the basis for this response. PMID- 8228258 TI - Murine recipients of fully mismatched donor marrow are protected from lethal graft-versus-host disease by the in vivo administration of rapamycin but develop an autoimmune-like syndrome. AB - We investigated the ability of the macrolide antifungal agent rapamycin (RAPA) to inhibit murine graft-vs-host disease induced across the MHC barrier. An optimum dose (1.5 mg/kg) given for 14 days beginning on the day of transplant (and then three times weekly until 1 mo) effectively and significantly (p < 0.001) protected 80% of irradiated B10.BR recipients of C57Bl/6 bone marrow/spleen grafts for over 90 days, whereas 80% of control mice died by day 37. Using a congenic model in which a mixture of Ly5.1+ bone marrow (T cell-depleted) and Ly5.2+ spleen cells allowed us to distinguish mature and immature cells, we found that RAPA inhibits the splenic expansion of mature donor-derived T cells in B10.BR recipients after bone marrow transplantation. In addition, phenotyping studies revealed that RAPA causes a massive reduction of immature CD4+CD8+ T cells in the thymus, indicating that RAPA probably interferes with maturation of immature CD3-CD4-CD8- T cells to CD4+CD8+ T cells. There was also a predilection toward development or intrathymic retention of the more mature CD3+CD4-CD8+ or CD3+CD4+CD8- cells in the thymus of long term survivors. These same observations were made in different experiments with mice given syngeneic bone marrow transplantation and RAPA. However, RAPA administration was associated with the occurrence of an autoimmune-like syndrome, consisting of ulcerative dermatitis, hepatic bile duct proliferation, and nondestructive lymphoid peribronchiolar infiltration of the lung. RAPA interfered with the deletion of potentially self reactive T cells that occurs in thymic development. The failure of clonal deletion was observed in allogeneic and syngeneic transplants given RAPA, although only the allografted mice experienced an autoimmune-like syndrome. Some, but not all, of the nondeleted V beta populations were functionally active. These new findings bear certain dissimilarities to the syndrome and lesions observed with cyclosporin A treatment, particularly in the observation of bile duct proliferation and ulcerative skin lesions. Nonetheless, because of the potent effect of RAPA in preventing lethal graft-vs-host induced across the MHC, further investigation of the immune consequences of this highly effective compound is warranted. PMID- 8228259 TI - Bullous pemphigoid and herpes gestationis autoantibodies recognize a common non collagenous site on the BP180 ectodomain. AB - Bullous pemphigoid (BP) and herpes gestationis (HG) are skin diseases characterized by subepidermal blisters and autoantibodies against two hemidesmosomal Ag, i.e., BP230 and BP180. Based on sequence analysis the BP180 Ag was predicted to be a transmembrane protein with a long extracellular collagenous domain. In the present investigation fusion proteins encompassing various segments of the BP180 Ag were expressed in a prokaryotic system and assayed by immunoblotting and immunoadsorption against a panel of BP, HG and control sera. One antigenic site, comprising 14 amino acids of the BP180 noncollagenous (NC) 16A domain, was shown to be recognized by 60% of BP sera and by 63% of HG sera tested. 73% (11/15) of BP sera and 100% (8/8) of HG sera reacted with at least one of three BP180 fusion proteins representing various portions of the NC16A domain. Immunoadsorption analysis identified this region of BP180 as an immunodominant site. Using an affinity purified rabbit antiserum raised against a recombinant form of BP180, this BP/HG autoantibody-reactive region was localized to the epidermal basal lamina immediately adjacent to the hemidesmosome. These findings confirmed the predicted type II transmembrane orientation of the BP180 Ag. Thus, the long, C-terminal collagenous domain of this basal keratinocyte protein projects into the basal lamina and may function as a site of interaction with an extracellular matrix component. It is proposed that autoantibodies directed against the well-defined antigenic site on the BP180 ectodomain may play an initiatory role in subepidermal blister formation in BP and HG patients. PMID- 8228260 TI - Clonal predominance of T cell receptors within the CD8+ CD45RO+ subset in normal human subjects. AB - Structural models for the TCR alpha/beta predict that the CDR1, CDR2, and CDR3 loops of both the alpha- and beta-chains contribute to specific interactions with the Ag/MHC complex. The CDR3 loops are constructed by joining events involving the V-(D)-J segments, and thus may vary in both sequence and length. We have developed a polymerase chain reaction assay to assess the length variation of the CDR3 loop in TCR derived from seven V beta segment families (V beta 2, V beta 3, V beta 4, V beta 9, V beta 14, V beta 16, and V beta 17). Peripheral blood T cells from 10 normal adults as well as five cord blood samples were studied. CD4+ and CD8+ T cells were analyzed separately. We observed extreme predominance of particular CDR3 lengths in half of the normal adults. These TCR were shown to be clonal by direct sequence analysis. This clonal dominance was found in the CD8+, CD45RO+ T cell population, and was observed in various V segment families. These patterns of TCR clonality were persistent over many months of observation in some individuals. In one subject, the new appearance of a predominant clone was associated with a booster vaccination for hepatitis B. These studies reveal a surprising degree of oligoclonality in the CD8+ cells of normal subjects which may be due to both environmental and genetic factors; the functional significance of persistent clonal dominance in the CD8 compartment remains to be determined. PMID- 8228261 TI - Recognition of human insulin in the context of HLA-DRB1*0406 products by T cells of insulin autoimmune syndrome patients and healthy donors. AB - Our recent study indicated that all the insulin autoimmune syndrome (IAS) patients had specific HLA class II alleles, the DRB1*0406, DQA1*0301, and DQB1*0302, which allowed T cells to proliferate when autologous APC were exposed to human insulin. The study implied that gene products of DRB1*0406, DQA1*0301, and/or DQB1*0302 may be involved in the presentation of human insulin to T cells. We therefore examined T cell response of healthy donors with different HLA phenotypes to human insulin using an autologous MLR system. The T cells from not only IAS patients but also healthy donors were able to proliferate after exposure of human insulin to autologous APC with DRB1*0406, DQA1*0301, and DQB1*0302 products. The class II molecules are considered to be involved in the recognition of human insulin by T cells. The proliferative response of T cells was completely blocked by anti-HLA-DR mAb and not by anti-HLA-DQ mAb or other mAb. Furthermore, human insulin-specific CD4-positive T cell clones were established from blast cells in autologous MLR of PBMC from two healthy donors with DRB1*0406 in the presence of human insulin. Using DRB1*0406-transfected L cells as APC, we confirmed that these T cells clones recognize human insulin in the context of gene products of DRB1*0406. These results provide the first evidence that HLA DRB1*0406 products act as the dominant restriction element for the presentation of human insulin to T cells, and suggest that this particular class II gene, HLA DRB1*0406, contributes to the development of IAS. PMID- 8228262 TI - Exacerbation of murine herpes simplex virus-mediated stromal keratitis by Th2 type T cells. AB - Corneal infection of susceptible mice with HSV-1 causes herpetic stromal keratitis (HSK), which serves as a model of human HSK. To study the properties of the T lymphocytes involved in HSK, susceptible mice were immunized with the synthetic peptide corresponding to the amino terminal of HSV-1-associated glycoprotein D (gD 5-23). A CD4+ long-term T cell line and a clone bearing V beta 8.2 TCR were derived from peptide-primed lymph node cells. These T cells recognize gD 5-23 peptide in the context of I-Ed and require CD4 and LFA-1 for Ag specific proliferation. Significantly, a truncated peptide gD 15-23 induced vigorous proliferation, indicating that these 9 amino acids constitute an epitope recognized by these T cells. The gD-specific T cells produced IL-4 and used it as the autocrine growth factor and hence belong to the Th2 subtype. Adoptive transfer of gD-specific Th2 cells into susceptible mice increased both the onset and severity of HSK after corneal HSV-1 infection. Injection of gD-specific Th2 cells without HSV-1 infection failed to cause eye damage. In addition, an irrelevant Ag-specific Th2 clone failed to induce similar tissue damage when the corresponding Ag was applied to the eye. These data indicate that the T cell mediated exacerbation of HSK in these studies is dependent on the specific recognition of gD after corneal HSV-1 infection. Finally, gD-specific Th2 cell transfer also rendered HSK-resistant mice susceptible for HSK, suggesting that the freedom from HSK in resistant mice may primarily be due to their inability to produce the pathogenic Th2 cells. The data collectively implicate an important role for Th2 cells in the induction of HSV-mediated keratitis in mice. PMID- 8228263 TI - Identification of human pre-T/NK cell-associated genes. AB - We have used a combination of subtractive cloning and differential screening techniques to identify genes preferentially expressed in early stages of human T/NK cell development compared with mature T and NK cells. A fetal liver-derived cytoplasmic (c) CD3+ membrane (m) CD3- clone, FL508, which expresses markers characteristic of pre-T and pre-NK cells served as a cell source for our cloning experiments. A cDNA library enriched for genes expressed in FL508 was constructed by removal of cDNA that hybridized to mRNA from a B cell line, JY. One-tenth of the resulting library of 5000 clones was screened by differential hybridization with cDNA probes from JY and a mature CD4+ T cell clone, B21. The relative expression levels of six selected clones were analyzed in 17 different cell/tissue types by semiquantitative polymerase chain reaction. Four of these clones are expressed at higher levels in thymocytes than in mature T or NK cells, and three clones are expressed at higher levels in fetal liver cells than in either mature T or NK cells. Partial and complete DNA sequence information suggests that these six cDNA correspond to previously unidentified genes. Genes identified in this study may be useful not only as markers for early stages of T/NK cell ontogeny, but also as tools for understanding novel developmental events. PMID- 8228264 TI - Molecular selection of human antibodies with an unconventional bacterial B cell antigen. AB - Unconventional Ag for B cells that are comparable to known superantigens for T lymphocytes have not been well characterized. However, the bacterial membrane protein, Staphylococcal protein A (SpA), has sites that interact with the Fab of many IgM, IgA, IgG, and IgE, and in recent reports we have provided evidence of VH restriction in Fab that bind SpA. To investigate the molecular basis for this Fab binding specificity, we have selected monoclonal Fab from a phage-display combinatorial Ig library, based on the ability to bind SpA. By this approach, in an unselected human polyclonal IgG Fab library, about 17% of antibody-expressing clones were found to bind SpA. SpA binding was completely restricted to Fab with VH3 H chains, and about 60% of VH3 Fab in the unselected library had SpA binding capacity. Analysis of 21 VH sequences and 6 VL sequences demonstrated that Fab that bind SpA use diverse VH3 genes, while the L chains derive from a variety of V kappa and V lambda gene families. By creation of antibodies with differential H L chain pairing, the global capacity to bind SpA was shown to be dictated by VH3 usage, but different L chain usage could result in up to a fourfold change in binding affinity. The apparent KD of the SpA binding by different Fab ranged from 2.5 x 10(-7) to > 10(-5) M. Furthermore, repeated rounds of in vitro panning selected for antibodies based on higher binding affinity. These data indicate that the pattern of VH family restriction of Ig reactive with SpA is comparable to known superantigens, and there is a hierarchy within the binding affinities of VH3 Fab based on V gene usage. PMID- 8228265 TI - Specific IgG autoantibodies are potent inhibitors of autoreactive T cell response to phytohemagglutinin-activated T cells. AB - PHA-activated human T cells express MHC class II molecules and have been shown to stimulate autoreactive T cells in autologous mixed lymphocyte reaction (T-T AMLR). We now demonstrate that normal human serum can dramatically suppress the proliferative response in T-T AMLR. The inhibitory factor was detected in the IgG fraction, and the activity was found to be mediated through the F(ab')2 portion of the IgG molecule, implying that the inhibitor could be specific. All homologous as well as autologous sera tested contained the inhibitory activity. Human serum was also found to suppress allogenic MLR stimulated by PHA-activated T lymphocytes but not allogenic MLR stimulated by nonactivated PBMC, indicating that the mechanism of inhibition is related to PHA activation of the stimulator cells. Moreover, pretreatment of the PHA-activated T lymphocytes with human serum resulted in a significant inhibition of T-T AMLR, as opposed to pretreatment of nonactivated PBMC, indicating that PHA-activated stimulators are functionally involved in the inhibition. Cytofluorometric analysis revealed that autologous IgG specifically binds to PHA-activated T lymphocytes and not to nonactivated CD3+ T cells. Furthermore, serum absorbed by PHA-activated T lymphocytes substantially lost its inhibitory activity whereas serum absorbed by nonactivated PBMC did not, suggesting that a surface molecule(s) expressed during activation of the stimulator cells is involved in the inhibition. The addition of human serum later in the culture period (> 3 days) resulted in a marked decrease in inhibition, implying that the presence of IgG in the early recognition phase of T T AMLR is essential for maximum inhibitory effect. These results raise the possibility that natural autoantibodies present in normal human IgG may play an important role in regulating immune response mediated by autoreactive T cells. PMID- 8228266 TI - Preparation of CD8bright and CD8dim lymphocyte populations using two positive selection methods in tandem. AB - Two positive selection methods were compared for the ability to capture both the bright and dim subsets of CD8 lymphocytes in mononuclear cell (MC) preparations from ten healthy individuals. The first method utilized anti-CD8-coated magnetic beads; captured cells were then recovered using a polyclonal sheep anti-mouse Fab reagent. At all bead: CD8 cell ratios tested (4:1, 8:1, 16:1), the selected cells were > 94% CD8+, and these CD8 cells were enriched for CD8bright cells (77-85%) when compared to CD8 cells in the starting MC preparation (68%). The second method utilized anti-CD8-coated culture flasks; captured cells were recovered by physical dislodgement. The recovered cells were > 90% CD8+, and these CD8 cells were modestly enriched for CD8dim cells (52%) compared to starting CD8 cells (32%). To further enrich for CD8dim cells, we used these two methods in tandem (n = 10). MC were first incubated with anti-CD8-coated magnetic beads (4:1 ratio) to obtain a CD8bright-enriched population (97% of all cells CD8+, 83% of all cells CD8bright). Uncaptured cells were incubated with anti-CD4-coated magnetic beads, and the uncaptured cells from this step were then placed in an anti-CD8-coated flask. The recovered flask-selected cell population was highly enriched for CD8dim cells (87% of all cells CD8+, 85% of all cells CD8dim). CD8 cells in the CD8bright population were 94% CD3+ and 6% CD16+, whereas those in the CD8dim population were 29% CD3+ and 66% CD16+. In proliferative studies, CD8bright cells were preferentially activated by immobilized anti-CD3, whereas CD8dim cells were preferentially activated by exogenous IL-2. In assays of natural killer activity, CD8dim cells were markedly more active than CD8bright cells. This method provides an alternative to cell sorting for obtaining enriched populations of CD8bright and CD8dim lymphocytes. PMID- 8228267 TI - Rapid transgene expression in lymphocyte and macrophage primary cultures after particle bombardment-mediated gene transfer. AB - A particle bombardment technique was used for gene transfer to human peripheral blood mononuclear cells, and murine splenocytes, thymocytes and peritoneal macrophages in primary culture. Significant expression of a luciferase marker gene was observed in these cell types within 8 h of gene transfer. Luciferase expression was readily detected in peritoneal macrophages 4 h after culture initiation and transfection. Same day determinations of transgene activity in fresh human peripheral blood mononuclear cell samples were feasible. Promoter preference and ballistic parameters were examined to optimize transgene expression. Up to 6% of bombarded human T lymphocytes expressed transgenic beta galactosidase activity. These results demonstrate that particle bombardment is an effective means for gene transfer and provides an attractive approach for rapid, quantitative analysis of transgene expression in various leukocyte primary culture systems. PMID- 8228268 TI - A comparison of the binding of biotin and biotinylated macromolecular ligands to an anti-biotin monoclonal antibody and to streptavidin. AB - A competitive enzyme immunoassay was used to study the binding of biotinylated macromolecular ligands and d-biotin to an anti-biotin monoclonal antibody and to streptavidin. Solid phase BSA-c-biotin competed with biotin or biotinylated macromolecular ligands in solution for receptor binding. The concentration of d biotin required to inhibit streptavidin binding to solid phase BSA-c-biotin by 50% was 11.5 pM. This streptavidin-biotin interaction was taken as having an affinity/avidity index of 100 and all other receptor-ligand interactions were calculated relative to this. The avidity indices calculated for streptavidin interactions with BSA-c-biotin and IgG-biotin were 17.6 and 6.6 respectively, whereas for anti-biotin the values for these ligands were 20.5 and 19.9 respectively. The interaction of anti-biotin with d-biotin had an affinity index of 0.001. Although streptavidin has the greatest binding affinity for d-biotin, its avidity for biotinylated ligands was considerably lower and comparable to that observed for anti-biotin-biotinylated macromolecule interactions. PMID- 8228269 TI - Kinetics of the autologous red cell agglutination test. AB - The kinetics of the autologous red cell agglutination test for detecting circulating antibodies to HIV-1 were studied. Two monoclonal anti-red blood cell antibodies (1C3/86 and 10F7MN) were used to construct Fab-peptide conjugates for the test. Both antibodies recognize glycophorin alpha on the surface of erythrocytes by immunoprecipitation or immunoblotting techniques. The number of binding sites, association and dissociation constants of 1C3/86 Fab and 10F7MN Fab' fragments were determined (n = 4.80 X 10(5) sites/erythrocyte, Ka = 0.43 X 10(7) M-1, Kd = 23 X 10(-8) M for 1C3/86, n = 4.66 X 10(5) sites/erythrocyte, Ka = 1.05 X 10(7) M-1, Kd = 9.5 X 10(-8) M for 10F7MN. The binding studies were performed under the same conditions as the autologous red blood cell agglutination test. When 0.9 microgram of anti-glycophorin Fab was added to 10 microliters of blood 0.25 microgram of 1C3/86 Fab was bound whereas 0.29 microgram for 10F7MN Fab' was bound. Antibody binding reached a plateau after 2 min and once bound did not exchange with unbound Fab over the time scale of the test. The binding of the anti-peptide antibody (cross-linking antibody) was also complete within 2 min. Addition of approximately 0.1 microgram of anti-peptide antibody gave half a maximal agglutination score. This is equivalent to 10 micrograms/ml circulating antibody. Under the agglutination test conditions, Fab peptide conjugate was bound to 14% of available glycophorin molecules. Half maximal agglutination occurred when approximately 1.1% of the bound Fab-peptide conjugates were cross-linked. A maximum agglutination score of four occurred in the presence of 1 microgram of anti-peptide antibody equivalent to 100 micrograms/ml circulating antibody whereas an agglutination score of 1+ was elicited by only 0.32 microgram anti-peptide antibody and involved the cross linking of approximately 160 glycophorin molecules per red cell. PMID- 8228270 TI - A simple and inexpensive high density dialysis tubing cell culture system for the in vitro production of monoclonal antibodies in high concentration. AB - This paper describes the construction and application of a low-cost roller bottle like culture appliance in which hybridoma cells can be cultivated in high density in dialysis tubing. The appliance facilitates the simultaneous culture of up to four cell lines yielding 50 ml culture volume of each. Samples for follow-up analysis of the cultures can easily be taken when needed through sample ports. In order to obtain high cell densities (at least 10(7) cells/ml), high cell viability (at least 50%) and high antibody yield (at least 1.0 mg/ml) the bottle is rolled at a speed of 4-6 rpm and is gassed continuously by a micropump driven by rechargeable NiCd batteries fixed to the culture flask. Depending on the individual properties of the hybridoma lines tested, the cells may be cultured for 1-2 weeks, and cell densities of up to 30 x 10(6) cells/ml with viabilities of approximately 50% and monoclonal antibodies in concentrations of up to 2.8 mg/ml may be obtained. In their properties the monoclonal antibodies produced by this in vitro procedure are indistinguishable from those prepared in the form of conventional stationary culture supernatant or of ascitic fluid. Specific antibody content is within the same range as in ascitic fluid. Consequently, the monoclonal antibodies can be purified in one step, e.g., by ion exchange chromatography from the culture supernatant. Therefore, the newly developed culture device and the culture method described is a useful alternative to ascites production in live mice. PMID- 8228271 TI - A simple technique for the rapid enrichment of class and subclass hybridoma switch variants. A 1000-fold enrichment in half the time, for half the cost. AB - Switching parental hybrids in vitro to downstream switch variant clones producing more desirable monoclonal antibodies (MoAbs) requires either labor intensive and time consuming subcloning techniques, or fluorescence activated sorting of the desired clones. We tested the hypothesis that enrichment of downstream switch variant clones might be achieved by selective lysis of upstream hybridoma cells followed by expansion of the enriched downstream clone. Using a parental hybridoma with surface and secretory IgM, we attempted to enrich downstream switch variant clones producing class (IgG) and subclass (IgG1 or IgG2a) MoAbs. Enrichment of downstream IgG, IgG1 and IgG2a MoAb-producing switch variants was achieved by single or repeated antibody-dependent, complement-mediated lysis of the upstream IgM-bearing parental hybridoma cells followed by limited subcloning. Two exposures of parental hybridoma cells to lysis followed by plating at 100 cells/well enriched the frequency of switch variants up to 1235-fold, enabling the development of IgG1 or IgG2a-producing subclones exhibiting high yield antibody production. Using this protocol, production time and costs were reduced by > 50% when compared to the standard technique. This novel technique for the rapid isolation and expansion of switch variant clones should be ideal for most laboratories, particularly those without access to cell sorting capabilities. PMID- 8228272 TI - Dot-blot immunodetection of antibodies against GM1 and other gangliosides on PVDF P membranes. AB - A simple and rapid assay for detection of antibodies against GM1 and other gangliosides (GM3, GM2, GD1a, GD1b, GT1b, GD3) is described. Purified gangliosides were applied individually in 1 microliter of methanol to polyvinylidene difluoride (PVDF) membranes. Anti-ganglioside antibodies in human sera were allowed to bind and were revealed with a second antibody coupled to peroxidase. The specificity of antibodies binding to gangliosides was confirmed using established techniques to detect anti-ganglioside antibodies such as immunostaining of gangliosides after high performance thin layer chromatography according to Derrington et al. (1989) and ELISA procedure according to Adams et al. (1991) or using the ability of cholera toxin beta subunit to remove GM1 bound antibodies. The dot-blot assay is the simplest and quickest method to run and it appears to be suitable for large routine screening detection of anti-ganglioside antibodies in sera of patients with neurological diseases. PMID- 8228273 TI - Accurate and rapid assessment of MHC antigen upregulation following cytokine stimulation. AB - Three human tumour cell lines, A431 (cervical), Scaber (bladder) and Fen (bladder), were studied using immunohistochemical staining (IC), radiobinding (RB), immunoprecipitation (IP), enzyme-linked immunosorbent assay (ELISA) and dot blot (DB) techniques in order to assess major histocompatibility antigen (MHC) induction in response to interferon-gamma (IFN-gamma). Induction of class II antigens by IFN-gamma was observed on all three cell lines using all techniques. Monoclonal antibody (Mab) staining showed that both Scaber and A431 lines were positive for intact class I (Mab W6/32), class I free heavy chain (Mab HC10) and beta 2 microglobulin (beta 2-m) (Mab BBM.1), while Fen cells were positive only with HC10. The IP technique demonstrated the presence of a 45 kDa band on precipitation of the Fen lysate with HC10 Mab, whereas no such band was observed when W6/32 was used. The DB technique confirmed the negative reaction with W6/32 and BBM.1 Mabs, while HC10 showed positive staining which was upregulated by IFN gamma. Transfection of the Fen cells with the beta 2-m gene resulted in the surface expression of fully assembled class I molecules. The DB technique showed upregulation of class I antigens following IFN-gamma stimulation, while RB detected no significant increase in cell surface expression (t test; p = 0.104). The binding values for transfected Fen cells before and after IFN-gamma stimulation were 2000 +/- 48 and 2161 +/- 156 cpm respectively. These results demonstrate that the DB technique facilitates an accurate assessment of cytokine induced antigens, corrected against a background of total cellular protein synthesis. The ease of execution, simplicity, non-radioactive nature and economy make it the method of choice for routine screening prior to the selection of suitable patients for cytokine therapy. PMID- 8228274 TI - Autoantibodies against topoisomerase I detected with the natural enzyme and overlapping recombinant peptides. AB - Antibodies against topoisomerase I (anti-topo I, anti-Scl-70) are regarded as a marker of systemic sclerosis. The various frequencies of anti-topo I detected in those patients depends at least in part on the test design and the kind of the antigen used. We therefore analyzed three overlapping recombinant topo I fragments (N-terminal, center and C-terminal part of the molecule) covering the full length of the enzyme for substitution of highly purified natural antigens (n topo I) in ELISA for antibody screening. 49 of 50 sera reacting with n-topo I in ELISA also recognized the recombinant C-terminal topo I fragment under identical test conditions, 37 sera recognized the recombinant center and two sera the recombinant N-terminal peptide. All sera reactive with the N-terminal and center peptide reacted with the recombinant C-terminus which therefore may substitute for the natural antigen. In immunoblot assays 92% (46/50) of the sera reacted with n-topo I and 86% (43/50) with the recombinant C-terminal peptide. Immunoblots therefore seem to be less sensitive for detecting anti-topo I antibodies than ELISA regardless the source of the antigen used. In a screening of 696 sera submitted for routine antibody tests the recombinant peptide ELISA on the other hand detected two sera which did not react with n-topo I in ELISA. Because of the high rate of agreement within the results obtained with the two antigens, n-topo I can be substituted by the recombinant peptide ELISA allowing better standardization and interlaboratory comparison of results. PMID- 8228275 TI - Comparison of two modified techniques for purifying blood eosinophils. AB - As there is much heterogeneity in the morphology and function of blood eosinophils, comparison of their properties between groups of subjects requires recovering the majority of these cells. In two currently used techniques to isolate eosinophils, blood granulocytes are processed either on Percoll gradients after an incubation of granulocytes with 10(-8) M N-formyl-methionyl-leucyl phenylalanine (fMLP) or on a magnetic cell sorter (MACS). In this study, these techniques were modified to increase the efficiency of eosinophil recovery. With the Percoll gradients, using 1.078 g/ml as the top gradient instead of 1.082 g/ml doubled the eosinophil recovery from 43 +/- 5.3% (mean +/- SEM) to 86.9 +/- 2.9%, without decreasing the purity (96.1 +/- 1.4% versus 96.2 +/- 0.9%). With a MACS, the neutrophils in granulocytes obtained on Ficoll-Paque (1.077 g/ml) instead of on Percoll gradient 1.082-1.094 g/ml, were tagged with anti-CD16 antibodies and eliminated by passing them through a magnetic field. When blood eosinophils of the same subjects were isolated using the two techniques, similar recovery and purity levels were obtained: Percoll gradients, 72.7 +/- 6.8% and 92.5 +/- 2.2%; MACS, 80.2 +/- 5.1% and 90.4 +/- 3.8%. Eosinophils isolated through the two techniques were also compared for their production of superoxide anion and leukotriene (LT) C4, with and without pre-incubation with cytokines interleukin 3, interleukin-5 and granulocyte-macrophage colony stimulating factor. The release of these products was similar between the two eosinophil preparations under all conditions tested except for interleukin-3 where eosinophils isolated with a MACS produced more LTC4. These results show that both techniques efficiently recover pure eosinophils. Furthermore, cell incubation with 10(-8) M fMLP did not enhance superoxide anion and LTC4 production nor modify the response to cytokines. The two modified techniques are therefore suitable for comparative studies of eosinophils from different groups of subjects. PMID- 8228276 TI - The use of chemiluminescence and the ELISA spot assay to identify and enumerate rare immunoglobulin switch variants. AB - The use of chemiluminescence and the ELISA spot assay for identifying rare immunoglobulin switch variants is described. The technique utilizes nitrocellulose membranes and allows rapid screening of a large number of cells. The number of spots can be recorded either manually or automatically by using a commercially available colony counting program. This modification of the ELISA spot assay makes it less labor intensive and time consuming and can be adapted for the search for rare cells secreting small amounts of Ig or other macromolecules. PMID- 8228277 TI - Processing proIL-1 beta decreases detection by a proIL-1 beta specific ELISA but increases detection by a conventional ELISA. AB - Recent investigations have noted that conventional IL-1 beta enzyme linked immunoassays (ELISA) may underestimate proIL-1 beta concentrations. In an attempt to circumvent this problem, we have devised a proIL-1 beta specific ELISA which sandwiches proIL-1 beta between a carboxy-terminus specific, capture antibody, and an amino-terminus specific, detection antibody. The amino-terminus specific antibody was generated against amino acids 3-21 of the intact proIL-1 beta molecule. This sandwich ELISA does not recognize mature 17 kDa IL-1 beta and is not inhibited by the coexistence of mature, 17 kDa IL-1 beta. We compared this proIL-1 beta specific ELISA (amino-terminus ELISA) to the conventional IL-1 beta ELISA (carboxy-terminus ELISA) on test samples that contained either proIL-1 beta or mature IL-1 beta. When purified proIL-1 beta is cleaved by increasing concentrations of IL-1 beta converting enzyme or porcine pancreatic elastase, there is a dose dependent increase in the signal from the conventional IL-1 beta ELISA and a concurrent decrease in signal from the proIL-1 beta specific ELISA. Furthermore, when the proIL-1 beta specific ELISA is used to quantify cellular sources of IL-1 beta, there is no detectable release of proIL-1 beta into the supernatants from either fresh blood monocytes or alveolar macrophages, despite detectable mature IL-1 beta. The cell associated compartment of IL-1 beta is largely proIL-1 beta and the relative amounts of IL-1 beta that remain intracellularly are quite large. Specifically, when assayed at 18 h, the proIL-1 beta ELISA detected 4.6 +/- 1.4 ng/ml per 10(6) monocytes and 13.8 ng/ml per 10(6) macrophages vs. 0.8 +/- 0.3 ng/ml/10(6) monocytes and 1.6 ng/ml/10(6) macrophages by conventional IL-1 beta ELISA. Thus, the 5-10-fold deficit in the detection of intracellular IL-1 beta by a conventional IL-1 beta ELISA, combined with the enhanced detection after enzymatic processing of proIL-1 beta, confirms that a proIL-1 beta specific ELISA is needed to accurately quantify proIL-1 beta. PMID- 8228278 TI - Detection and quantification of secreted soluble Fc gamma RIIA in human sera by an enzyme-linked immunosorbent assay. AB - Fc gamma RIIA can be produced in a soluble form that contains both the extracellular and intracellular regions of the receptor, due to an alternative splicing of the transmembrane domain-coding exon. We have developed an enzyme linked immunosorbent assay (ELISA) that permits the specific detection and quantification in human sera of this secreted soluble Fc gamma RIIA. It uses the monoclonal antibody (MAb) IV.3 as capture antibody and rabbit polyclonal IgG directed against the intracellular region of Fc gamma RIIA as detector antibodies. The enzymatic reaction was amplified using an NADH/NAD+ amplification system. As little as 0.8-1.5 ng/ml (20-38 pM) of purified recombinant secreted Fc gamma RIIA could be detected. The serum levels of secreted sFc gamma RIIA ranged from 0 to 30 ng/ml in sera from 51 healthy donors. The mean value was 11.9 ng/ml +/- 6.55 (297 pM +/- 163) and the median value was 10.6 ng/ml (265 pM) (range: 0 764 pM). PMID- 8228279 TI - A capillary electrophoresis-based assay for the binding of Ca2+ and phosphorylcholine to human C-reactive protein. AB - Affinity capillary electrophoresis was performed to quantitate the binding of Ca2+ and phosphorylcholine to human C-reactive protein (CRP). The assay requires no modifications of any of the molecules involved, uses minuscule amounts of protein (8.5 x 10(-15) mol per analysis, i.e., less than 1 pmol for 15 triplicate data points), and the binding could be examined under conditions of physiological ionic strength and pH. The values for the dissociation constants obtained here (KD = 59 microM for Ca(2+)-CRP and 18 microM for the phosphorylcholine-CRP interaction) were in close agreement with previous studies using gel filtration and equilibrium dialysis. As long as one of the reactants can be detected and recovered quantitatively in the capillary electrophoresis system, the method is generally useful to study interactions where complexed molecules display an electrophoretic mobility that is different from that of unbound molecules and where the rates of association and dissociation are sufficiently fast. PMID- 8228280 TI - Analysis of lymphocyte aggregation using digital image analysis. AB - We present the development and testing of a novel assay of lymphocyte adhesion based on time-resolved morphological measurements of intercellular aggregation. Homotypic lymphocyte aggregation is induced according to various protocols and monitored for several hours using video microscopy and time-lapse recording. Digital images of the aggregating cell population are acquired and analyzed to obtain the size distribution and the shape of cell aggregates. By following the temporal evolution of the size distribution of aggregates, the rates of aggregation events can be accurately quantified and compared. In addition, an analysis of the two- and three-dimensional structures of the aggregates using appropriately defined shape factors allows comparisons of mechanical binding strengths and cytoskeletal activity. To demonstrate the capabilities of the assay, we present results from a series of aggregation experiments with Jurkat cells treated with 33B6, 19H8, IC9, and 20E4 monoclonal antibodies. These monoclonal antibodies bind to various epitopes of known adhesion molecules and induce aggregation phenomena that proceed at different rates. Our results show that the assay has small repeatability error and is sensitive enough to compare aggregation events induced through distinct molecular epitopes. Used in conjunction with current biochemical detection assays and adhesion pathway modulation experiments, the developed assay will facilitate the study of cellular adhesion and aggregation mechanisms. PMID- 8228281 TI - Carrier determination for X-linked agammaglobulinemia using X inactivation analysis of purified B cells. AB - We report the development of a relatively quick and simple method for the assessment of X inactivation status for carrier determination in families affected by X-linked agammaglobulinemia (XLA). This method utilises an immunomagnetic separation technique for B cell purification and a polymerase chain reaction (PCR) based assay for the determination of methylation status at the androgen receptor (AR) gene locus to assess whether X inactivation is random or non-random at this locus. We report the results we have obtained using this assay to investigate females known to be carriers of various X-linked immunodeficiency disorders. In addition, we investigated four females from different families affected by XLA, two of whom were of unknown carrier status, and we discuss the results obtained with this and other X-inactivation assays. A similar assay has recently been described by Allen et al. (1992) and applied to members of one family affected by XLA. PMID- 8228282 TI - Purification and properties of immunotoxins containing one vs. two deglycosylated ricin A chains. AB - A method for the preparation of immunotoxins (ITs) containing one and two molecules of deglycosylated ricin A chain (dgA) bound to each molecule of anti human CD22 mouse IgG1 monoclonal antibody is described. The cross-linking of antibody and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-alpha methyl-(2-pyridyldithio) toluene. The two ITs were purified by successive chromatographies on Blue-Sepharose, Sephacryl S-200, Blue-Sepharose with NaCl gradient and Sephacryl S-300. The IT with two dgAs was seven-fold more cytotoxic to CD22+ Daudi cells than that containing one molecule of dgA. PMID- 8228283 TI - Rapid immunofiltration assay of Newcastle disease virus using a silicon sensor. AB - A rapid nonradioactive sandwich immunoassay which utilizes biotin-streptavidin mediated filtration capture of immune complexes in conjunction with a silicon sensor was developed for the detection of virus. Using purified Newcastle disease virus as a model, the lower limits of detection (LOD) were determined for a number of immunoassay configurations employing both monoclonal and polyclonal antibodies. The LODs ranged from 1.3 ng/ml (sample volume of 100 microliter) for an incubation of 60 min to 400 ng/ml for a 1 min incubation. The sandwich immune complexes were formed from one-step incubation of antibody and antigen. No 'hook' effects were observed over a wide range of analyte concentrations. The assays were easy to perform and required a total time equal to the incubation period plus about 5 min. The assay format is suitable for virus, bacteria and protein antigens. New assays can be developed and optimized readily, often within 1 day. PMID- 8228284 TI - Measurement of kinetic binding constants of viral antibodies using a new biosensor technology. AB - Association (ka) and dissociation (kd) rate constants of three monoclonal antibodies raised against tobacco mosaic virus were determined using a biosensor technique based on surface plasmon resonance (BIAcore, Pharmacia). Dissociation rates were constant over the 4-400 nM antibody concentration range whereas apparent association rates decreased over this range probably due to an increased saturation level of the antigen. Affinity constants K calculated from the ratio of ka/kd were in reasonable agreement with values obtained under equilibrium conditions by two standard methods based on enzyme immunoassay. PMID- 8228285 TI - Ca2+ response in single human T cells induced by stimulation of CD4 or CD8 and interference with CD3 stimulation. AB - A Ca2+ imaging method has been used to demonstrate simultaneously the magnitude and time course of the increase in the intracellular Ca2+ concentration ([Ca2+]i) in 10-30 individual human peripheral T cells following stimulation by anti-CD4 or anti-CD8 monoclonal antibody (MAb) as well as anti-CD3 MAb. The rise in [Ca2+]i began within 10 s of the introduction of the MAb and reached a peak of 240 nM (mean of 73 cells) in 20-40 s. The peak was followed by a slow decrease persisting for 6-8 min. Comparing Ca2+ responses in the presence and absence of external Ca2+, the rise in [Ca2+]i was found to be caused by both transient intracellular Ca2+ release and a long-lasting Ca2+ influx from outside the cell. Cross-linking of CD4 or CD8 using anti-IgG antibody augmented the response in individual cells, as seen in the higher peak (365-390 nM) and the longer duration (over 10 min). Simultaneous stimulation of CD3 and CD4 did not cause a summation of Ca2+ responses but caused a suppression in the CD3-mediated Ca2+ response. The results support the view that CD4 and CD8 play a role in signal transduction for T cell activation and that the CD4-derived signal interferes with the CD3-derived signal at some stage in the signalling pathway causing the Ca2+ response. PMID- 8228286 TI - Human IgG preparations isolated by ion-exchange or protein G affinity chromatography differ in their glycosylation profiles. AB - IgG from patients with rheumatoid arthritis (RA) is abnormally glycosylated in the Fc region, with sialic acid and galactose levels lower than normal. Protein G and DEAE purify populations which are differentially glycosylated. Significantly increased exposure of sialic acid was detected in normal IgG compared with that of RA IgG when ion exchange was used to prepare samples. However, when the same samples were prepared using protein G, no difference in the detection of sialic acid was seen between the two groups. When examining the heavy chain of IgG, more sialic acid, galactose and N-acetylglucosamine were detected in DEAE purified IgG compared with that prepared by protein G Detection of sialic acid and N acetylglucosamine was also increased on light chains from IgG prepared by ion exchange chromatography. Since this occurs notably on rheumatoid light chains it would appear that this arrangement would contribute to the overall glycosylation changes in IgG. In the case of molecules lacking galactose the discrimination between the RA and normal IgG is significantly improved when ion exchange chromatography is used. Since differentiation between disease and normal groups relies on the purification technique used, we recommend that more than one method is employed before undertaking an analysis of glycosylation changes. PMID- 8228287 TI - Rapid flow cytometric assay for the assessment of natural killer cell activity. AB - A new assay using flow cytometry has been established to assess natural killer (NK) lytic activity with common bench top instrumentation. This assay uses a cyanine membrane dye to stain live K562 target cells and an iodide nuclear dye to evaluate dead cells, and provides a method of reliably separating target and effector cell populations. Effector cells remain unstained (fluorescent negative) throughout the procedure. The damaged pre-labeled targets appear doubly stained as their membranes become permeable to the nuclear stain during incubation. Percent cytotoxicity of various effector:target cell ratios is discerned using flow cytometric analysis after a 2 h incubation in this new assay, as compared to 4 h with the 51Cr-release 'gold standard' assay for cell-mediated cytotoxicity. Comparisons of normal individuals tested in parallel with the fluorescent dyes and the 51Cr-release assay have shown direct correlations. This new two-color flow cytometric technique has proven to be uncomplicated and reproducible when used in the clinical setting. PMID- 8228288 TI - Covalent coupling of a steroid to microwell plates for use in a competitive enzyme-linked immunosorbent assay. AB - The covalent coupling of a model steroid, 17 alpha-hydroxypregnenolone, to the wells of the microtiter plate, CovaLink NH, for use in a competitive enzyme linked immunosorbent assay is described. This plate has secondary amino groups bound to its surface. A carboxylated derivative of the steroid was coupled to the amino group to form an amide bond in a single step using a water-soluble carbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (10 mM) as coupling reagent in the presence of N-hydroxysuccinimide (1 mM). After carrying out a competitive immune reaction, antibodies bound to immobilized steroids were estimated by means of a second antibody-enzyme conjugate. The non-specific background was reduced with blocking agents which did not interfere with the immune reaction between antibodies and the steroids coupled to the plastic surface. The following two procedures were effective for this purpose: pretreatment of wells with 0.01% Tween 20 solution followed by 0.5% bovine serum albumin in phosphate buffered saline, and addition of 0.01% Tween 20 to the assay buffer. With this method, the preparation of steroid-enzyme conjugates is unnecessary and optimization of conditions for ELISA procedures can be achieved in a simple manner. PMID- 8228289 TI - Analysis of active antibody concentration. Separation of affinity and concentration parameters. AB - Antibody binding to surfaces with differing amounts of immobilised antigen was measured in a biosensor system using surface plasmon resonance detection. Binding rates obtained during the initial binding phase on high density antigen surfaces were proportional to antibody concentration and independent of antigen-antibody affinity. One antibody calibration curve covering the range from 0.5 to 160 nM (0.08-25 micrograms/ml) antibody was valid for IgG antibodies with different antigen specificities. To illustrate the use of this methodology active antibody concentrations were analysed in culture media and in rabbit serum. PMID- 8228290 TI - The concentrations of glutamine and ammonia in commercially available cell culture media. AB - The amino acid glutamine is an essential nutrient for cells in culture. In aqueous solutions such as liquid culture media, glutamine spontaneously decomposes into ammonia. In this study, we examined the toxicity of ammonia for two different cell lines. In mouse hybridoma cell cultures, viable cell counts were reduced at exogenous ammonia concentrations of 1000 microM. In the human promyelocytic cell line however, viable cell counts were shown to be reduced at exogenous ammonia concentrations of 300 microM. Next, we determined ammonia and glutamine levels in 11 commercially available media on the day of delivery. It was found that all media contained significantly less glutamine than prescribed. Ammonia was found in all media with concentrations ranging up to 1000 microM. Storage at both 4 degrees C and 20 degrees C caused a further degradation of glutamine and significant accumulation of ammonia in all media. The degradation curves of the various media were used to calculate the first order degradation constant k, which can be used to determine the kinetics of the spontaneous decomposition in culture media. These results suggest that precautions must be taken to avoid the deterioration of commercially available culture media, because of the decay of glutamine. Long storage times lead to a rapid decay of glutamine and an accumulation of the toxic degradation product ammonia. PMID- 8228291 TI - Epidemic cholera due to a novel strain of V. cholerae non-01--the beginning of a new pandemic? PMID- 8228292 TI - Prevalence in Singapore of enterococci with high-level resistance to aminoglycosides and comparison of three methods for their detection. AB - The first study of the prevalence in Singapore of enterococci with high-level resistance to aminoglycosides has been made in the National University Hospital. From March to June 1992, 225 isolates were studied by means of three methods: (1) use of brain-heart infusion (BHI) agar incorporating the antibiotic; (2) microdilution of the antibiotic in broth; and (3) use of high content antibiotic discs. The BHI agar was the reference method. Of the total, 49 (22%) isolates were resistant to gentamicin, 86 (38%) to streptomycin, 82 (36%) to kanamycin and 93 (41%) to one or more of the agents. Results of the disc diffusion method correlated completely with the BHI agar method. The broth microdilution method showed a 100, 90 and 96% sensitivity in relation to gentamicin, streptomycin and kanamycin, respectively, and a specificity of 100% for all three aminoglycosides. Statistical significance (P < 0.01 to P < 0.001) was demonstrated in (1) a greater incidence of enterococci with high-level resistance to aminoglycosides among blood isolates; (2) gentamicin resistance predicting kanamycin high-level resistance; and (3) more resistance to gentamicin, streptomycin and kanamycin among Enterococcus faecium isolates compared to those of Enterococcus faecalis. Beta-lactamase activity was not detected. PMID- 8228293 TI - Blockage of urethral catheters by bacterial biofilms. AB - We report the case of a 63-year-old woman whose indwelling urethral catheter became blocked regularly at 4-5 day intervals over a period of 10 weeks. 'Worm like' structures 25-30 cm in length were found either in the catheter, completely occluding the lumen, or in the drainage tube thereby blocking the valve of the drainage bag. Electron microscopy showed that these structures were composed of bacteria, while culture revealed them to be mixed communities of Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecalis and Proteus mirabilis. Following treatment with ciprofloxacin, catheter drainage continued freely for a further period of 10 weeks. PMID- 8228294 TI - Use and costs of antimicrobial drugs in medical wards of a central hospital in Malawi. AB - Economic constraints experienced by many African countries make it vital that reliable information is collected about the cost of patient care in health institutions. Case files and treatment charts of 509 patients admitted to non-fee paying medical wards of Queen Elizabeth Central Hospital, Blantyre, Malawi for the month of September 1992 were analysed in order to determine the amount and costs of drugs and antimicrobial agents given to inpatients. Infectious diseases accounted for 76% of all admissions. A total of 59 different drug items were used (cost U.S. $732). In all, 18 antimicrobial agents were given (31% of all drug items) at a cost of U.S. $615 (84% of all drug costs). Calculated costs of antimicrobial agents for each patient and for annual admissions (10325 in 1992) were U.S. $1.21 and U.S. $12,558 respectively. Although the present study has its limitations, it provides a rational basis for planning departmental drug budgets. PMID- 8228295 TI - Immunisation of the preterm baby. AB - The study was designed to quantify the preterm baby's response to routine childhood immunisations. A total of 69 preterm babies were put into two groups according to their gestational age: less than 32 weeks and between 32 and 35 weeks. Within each group the babies were randomly placed into one of three schedules for immunisation with diphtheria, tetanus, pertussis and oral poliomyelitis vaccines at 3, 4 and 5 months; 3, 4, 5 and 18 months; and 3, 4 and 10 months. Antibodies were measured before immunisation, one month after the third vaccine dose and at age 19 months. Of these children, 30 had further antibody estimations when they were between 4 and 5 years of age. One month after the third immunisation and at 19 months of age all infants, independent of gestational age, had adequate immunity to the four infections. Children sampled before and after their pre-school booster, with few exceptions, showed persistence of protective antibody and an enhanced response to the booster dose respectively. No correction needs to be made for prematurity when initiating routine immunisation in premature infants. PMID- 8228296 TI - Family study of women showing development of antibody to human T-cell leukemia virus I and assessment of the risk of vertical transmission of the virus to their children. AB - When pregnant women were tested for antibody to human T-cell leukemia virus-I, some were found to be positive although they had been negative during the previous pregnancy. In these women, HTLV-I infection was found from pedigree studies to have been acquired from their mothers rather than from their husbands. Furthermore, some of them had apparently remained HTLV-I antibody-negative for long periods after infection. When the breast-fed children of these women, in whom HTLV-I was acquired from their mothers but who were in an HTLV-I antibody negative state, were also examined for evidence of HTLV-I infection, none was found. PMID- 8228297 TI - Antimicrobial susceptibility and serotype distribution of Streptococcus pneumoniae from Rwanda, 1984-1990. AB - A total of 383 clinical isolates of Streptococcus pneumoniae, obtained from an equal number of patients in Kigali, Rwanda, was tested for resistance to penicillin G with a 1 microgram oxacillin disc. Of these isolates, 99 (25.8%) showed reduced zones of inhibition. By means of an agar dilution method, 21% all isolates were confirmed as relatively resistant (MIC > or = 0.12- < or = 1.0 mg/l) strains of Streptococcus pneumoniae (RRSP). A high degree of resistance to penicillin G (MIC > or = 2 mg/l) was not observed. Resistance to chloramphenicol (MIC > or = 8 mg/l) was found in 31% RRSP and in 6% penicillin susceptible strains (PSSP). Doxycycline resistance was common in both RRSP and PSSP strains. All isolates remained fully susceptible to erythromycin. Children more often harboured a strain giving a reduced inhibition zone than did adults (74/230 versus 25/153; P = 0.0005). A total of 32 serotypes or serogroups were identified, seven of them relating to 64.8% all isolates typed. Of all the isolates 84% belonged to a serotype represented in the 23-valent vaccine or to a cross-reacting serotype. Serotype 25, not included in the vaccine, accounted for 10.7% typed isolates from adults but only for 2.0% typed isolates from children. Results of susceptibility testing and clinical experience suggest that penicillin G, ampicillin and chloramphenicol should not be used alone as empirical treatment for pneumococcal meningitis in patients in Rwanda. PMID- 8228298 TI - Brucella peritonitis. AB - We report the case of a patient with peritonitis caused by Brucella melitensis who also had chronic liver disease. At first the patient was treated for bacterial peritonitis but when a lymphocytic ascites was aspirated antituberculosis chemotherapy was given. However, the serological tests for brucellosis were strongly positive and subsequently B. melitensis was isolated from ascitic fluid. PMID- 8228299 TI - Phialophora richardsiae endocarditis of aortic and mitral valves in a diabetic man with a porcine mitral valve. AB - We report a case of Phialophora richardsiae endocarditis involving the aortic and a porcine mitral valve in a 52-year-old non-insulin dependent diabetic. Mitral valve replacement had been performed 4 years earlier. We believe this to be the first case of Phialophora richardsiae endocarditis to be reported. PMID- 8228300 TI - Treatment of recalcitrant actinomycosis with ciprofloxacin. AB - We describe a case of actinomycosis with several unusual features. The patient has been under medical supervision for more than 20 years; the disease has spread in an atypical manner and did not respond to standard therapy. Finally, there has been an unexpected response to a prolonged course of a member of the quinolone group of antibiotics. PMID- 8228301 TI - Hypoadrenal crisis caused by disseminated histoplasmosis. AB - We report a case of disseminated histoplasmosis presenting with acute renal impairment due to adrenal insufficiency. The finding of bilateral adrenal gland enlargement on abdominal ultrasound examination led to a diagnostic CT-guided adrenal biopsy. Prolonged therapy with amphotericin and itraconazole resulted in a clinical cure, although the patient still requires adrenal replacement therapy. PMID- 8228302 TI - Increasing incidence of cryptococcosis in the United Kingdom. AB - The incidence of infection with Cryptococcus neoformans in the United Kingdom during the years between 1953 and 1981 is compared with that between 1982 and 1991. The patients were those from whom samples were submitted to the PHLS Mycological Reference Laboratory (MRL), or those in whom the disease was confirmed elsewhere in the United Kingdom and reported to PHLS Communicable Disease Surveillance Centre (CDSC). In all, 83 cases were identified between 1953 and 1981 and 322 between 1982 and 1991, 201 of which were known to be HIV associated. The incidence of infection with Cryptococcus neoformans has increased four-fold in the last decade. It is an increasing cause of infection in immunosuppressed patients, most notably those with HIV infection. Currently, 4.0% patients with AIDS in the United Kingdom are known to have developed cryptococcosis. PMID- 8228303 TI - Surveillance of measles in Scotland 1983-1991; a comparison of statutory notification and laboratory reporting. AB - The epidemiology of measles in Scotland for the period 1983-1991, based on statutory notification and laboratory reporting, is described. There were 34,823 notifications compared to only 690 laboratory reports. This suggests that for less than 1 in 50 notifications of measles was the clinical diagnosis confirmed by laboratory testing. Both sets of data show a declining incidence of measles (in keeping with rising rates of immunisation) but a statistically significant shift in the age distribution of cases towards the over 15 years age group. There were differences, however, between the two sets of data in the sex ratio within various age groups. In particular, among notified cases, there was a highly significant but unexplained preponderance of males in the over 15 years age group (59.7% vs. 40.3%, P < 0.001). Discrepancies between the two sets of data were also evident from comparisons among Health Boards. Possible sources of bias are suggested for each set of data. The relevance of this surveillance to the current debate about the need for a two stage Measles/Mumps/Rubella (MMR) immunisation policy is briefly discussed. In order that this debate be reliably informed, it is highly desirable that the clinical diagnosis in each case of apparent vaccine failure is confirmed by laboratory testing. PMID- 8228304 TI - Erysipelothrix endocarditis affecting a porcine xenograft heart valve. PMID- 8228305 TI - Cutaneous abscesses caused by Salmonella enteritidis: an unusual presentation of salmonellosis. PMID- 8228306 TI - Relapsed infection due to Listeria monocytogenes confirmed by random amplified polymorphic DNA (RAPD) analysis. PMID- 8228307 TI - Recurrent coagulase-negative staphylococcal septic arthritis. PMID- 8228308 TI - Isolation of a Leuconostoc species from a retroareolar breast abscess. PMID- 8228309 TI - Streptococcus agalactiae meningitis in a previously healthy adult. PMID- 8228310 TI - High prevalence of serum antibodies to Verotoxins 1 and 2 among healthy adults in Japan. PMID- 8228311 TI - Ciprofloxacin-induced thrombocytopenia. PMID- 8228312 TI - Legionella pneumonia in a calf. PMID- 8228313 TI - Recurrent urinary tract infection with haematuria caused by Moraxella (Branhamella) catarrhalis. PMID- 8228314 TI - The present and future role of the consultant physician with an interest in infection. Council of the British Society for the Study of Infection. AB - The Council of the British Society for the Study of Infection (BSSI) has considered the present and future role of the Consultant Physician with an interest in infection. The BSSI is anxious to set standards and improve the quality of care delivered to patients with infection; to give advice on the prevention and management of infection and to provide such services economically to assist Health Managers in the purchase of infection-related medical services appropriate to the needs of the community they serve. New approaches to the care of infected patients are reviewed where the Clinical Specialist, Microbiologist and Consultant in Communicable Disease Control (CCDC)--in Scotland, Consultant in Public Health Medicine (CPHM)--work together in a team. As the U.K. moves closer to its European partners in 1992, it seems timely for each specialty to make a clear statement about its training programme and the requirements expected of a fully trained Consultant in each discipline. Comparisons between the ratio of Consultants with a special interest in infection and the population are made, where known, between U.K. and other European countries. A model job description for the U.K. clinical specialist is therefore being made available, the present inadequate numbers of consultants are stated and recommendations made to expand urgently the number of Consultant Physicians with an interest in infection. PMID- 8228315 TI - The consultant physician with an interest in infection--model job description. Council of the British Society for the Study of Infection. PMID- 8228316 TI - Purification and amplification of DNA from Penaeus monodon-type baculovirus (MBV). AB - The purpose of this paper was to purify and amplify the DNA fragment of Penaeus monodon-type baculovirus (MBV). Using 30-50% caesium chloride gradients, MBV virions and occlusion bodies with density parameters of 1.28-1.29 and 1.32-1.33 g/ml, respectively, were purified. Two oligonucleotide primers have been successfully designed and utilized for the amplification of a DNA fragment of MBV. After 35 amplification cycles of the MBV DNA fragment, a large amount of amplified product with an approximate molecular weight of 600 bp was obtained. This is the first successfully published work on the amplification of MBV using the polymerase chain reaction (PCR). Using the same primers, DNA extracted from MBV noninfected P. monodon, P. japonicus, and P. orientalis had a negative PCR response. However, a positive PCR response was obtained from DNA extracted from MBV-infected postlarval P. monodon. DIG-dot blot hybridization technique using PCR product obtained from the present study as a probe further confirmed that the product is originated from a portion of MBV polyhedrin gene. It is also suggested that PCR product may be beneficial for an accurate and early diagnosis of MBV infection in larval shrimp. PMID- 8228317 TI - Expression of full-length and truncated forms of crystal protein genes from Bacillus thuringiensis subsp. kurstaki in a baculovirus and pathogenicity of the recombinant viruses. AB - Full-length and truncated forms of the crystal protein gene cryIA(b) derived from Bacillus thuringiensis subsp. kurstaki HD-1 and full-length cryIA(c) gene of B. thuringiensis subsp. kurstaki HD-73 were introduced into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus, in place of the polyhedrin gene. All gene constructs were expressed at high levels in insect cells and insects upon infection with the recombinant viruses. The protein products were shown to be biologically and immunologically similar to the natural crystal protein. The expressed proteins formed crystals (in insects) up to 10 times bigger (in length) than their bacterial counterpart. The LT50 values for recombinant viruses were not significantly shorter than wild-type virus. PMID- 8228318 TI - Protection from ultraviolet irradiation by melanin of mosquitocidal activity of Bacillus thuringiensis var. israelensis. AB - A process for production, isolation, and purification of melanin produced by the fermentation of Streptomyces lividans 66 harboring a recombinant plasmid pIJ702 bearing tyrosinase gene has been developed. The efficacy of melanin in the protection of mosquito larvacidal activity of Bacillus thuringiensis var. israelensis against uv light has been studied. Results obtained by the live cell counts and the bioassay of residual mosquitocidal activity of B. thuringiensis var. israelensis after exposure to uv radiation showed that melanin is an excellent photoprotective agent. PMID- 8228319 TI - Life cycle of a new species of Duboscqia (Microsporida: Thelohaniidae) infecting the mosquito Anopheles hilli and an intermediate copepod host, Apocyclops dengizicus. AB - A new species of Microsporida, Duboscqia dengihilli, was found infecting larvae of the mosquito Anopheles hilli in northern Queensland, Australia. Laboratory experiments showed that binucleate spores formed within infected female mosquitoes were responsible for transovarial transmission to the next generation. Sporogony within the larval fat body was initiated by two diplokarya, one at each end of the cell, which undergo meiosis within a single sporophorous vesicle to form 16 meiospores. These spores are responsible for horizontal transmission to the copepod Apocyclops dengizicus. The microsporidium is transmitted back to the mosquito host via uninucleate pyriform spores formed within the copepod host which are infectious to larvae of A. hilli. The meronts within larvae infected by horizontal transmission ultimately develop into the binucleate spores within adult females to complete the life cycle. Thus, the development of this microsporidium involves vertical transmission between successive mosquito generations and horizontal transmission between mosquitoes and copepods similar to the life cycles of Amblyospora and Parathelohania. PMID- 8228320 TI - Phylogenetic interrelationships among baculoviruses: evolutionary rates and host associations. AB - The phylogenetic relationship of several baculovirus polyhedrin genes was investigated. Alignments of 18 polyhedrin gene amino acid sequences from which 12 were also available as DNA sequences were constructed based on positional homology and trees were calculated using character state and distance methods. Our results indicate that the Hymenopteran NsSNPV diverged from the Lepidopteran baculovirus before the separation of the NPV from the GV and subsequent radiation of these Lepidopteran groups. Moreover, this analysis indicates that lepidopteran NPV are clustered in at least two groups with distinct evolutionary rates. Group I includes the AcMNPV, BmMNPV, BmSNPV, OpMNPV, AgMNPV, and possibly the GmMNPV and AsMNPV. Group II includes the MbMNPV, PfMNPV, SfMNPV, SeMNPV, and possibly the OpSNPV. This grouping is also supported by a tree constructed from sequence data from the 5' untranslated leader region of the polyhedrin gene. Furthermore, the polyhedrin-based phylogeny was analyzed for its congruence to the genomic organization and other biological characteristics of the baculoviruses. This study shows that the SNPV and MNPV morphotypes have to be taken with caution as classificatory traits since they appear to be unpolarized. In addition, the comparison of the baculovirus phylogeny with the Lepidoptera phylogeny confirms the assumption that the evolutionary history of the baculoviruses is not entirely in agreement with the taxonomic organization of the Lepidoptera, implying that the common designation of baculoviruses based on the hosts may not be adequate for baculovirus classification since it does not reflect the genetic relatedness of the viruses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228321 TI - The antimicrosporidial activity of albendazole. AB - The antimicrosporidial activity of albendazole was tested on Nosema bombycis in vitro in Spodoptera frugiperda cells and in vivo in Heliocoverpa zea larvae and pupae. Significant reductions in the percentage of infected S. frugiperda cells were obtained using a concentration of 5.3 micrograms/ml albendazole in tissue culture medium but recrudescence occurred after the drug was withdrawn from the cultures. Significant reductions in the number of spores harvested from 6th instar larvae or pupae were obtained when doses of 0.2 to 4.0 mg were incorporated into the diet but, with the lower doses, some resurgence of infection occurred in pupae after cessation of drug intake. Established infections were almost eliminated from 6th-instar larvae and pupae after consumption of 2 or 4 mg albendazole and infections were not established at all when 4 mg was consumed concurrently with the infective spores. Even at the highest dose albendazole had no deleterious effect on the growth and viability of H. zea. Clumped chromatin in the nuclei of meronts, revealed by electron microscopy, reflected the selective anti-tubulin activity of albendazole and there was massive disorganization of sporogonic development. PMID- 8228322 TI - Human herpesvirus 6 is present in lesions of Langerhans cell histiocytosis. AB - Langerhans cell histiocytosis (LCH) is a disease characterized by Langerhans cell infiltration of skin and bone, with its most severe form manifested by multifocal infiltration of many organs. The etiology is unknown, although viral infection has been proposed as a potential pathogenic factor. Human herpesvirus 6 (HHV-6), a recently described member of the human herpesvirus family, has been associated with atypical or malignant lymphocytic processes, and immune disorders. Based on these observations, we suspected that HHV-6 may play a role in the pathogenesis of LCH. Lesional tissue of 30 patients with LCH was retrospectively examined for the presence of HHV-6 by using the polymerase chain reaction. Tissue specimens from 63 patients with other benign and malignant histiocytic and lymphocytic diseases served as controls. In addition, all specimens were examined with control primers specific for herpes simplex virus (HSV). HHV-6 DNA was detected in lesions of 14 of 30 patients with LCH (47%). On clinical subgroup analysis, HHV-6 DNA was found in 10 of 16 patients with extraosseous disease (63%) and in four of 14 patients with disease limited to bone (29%). In each case, the prevalence of HHV-6 in LCH lesions was statistically significant, when compared to the control population. HSV DNA was not found in any of the LCH or control specimens. Although the presence of a virus alone does not establish a causal role in the disease, it supports the possibility of an etiologic relationship. From this study, we emphasize the need for further investigation of the potential HHV-6-mediated pathogenesis of LCH. PMID- 8228323 TI - Direct effects of cutaneous neuropeptides on adenylyl cyclase activity and proliferation in a keratinocyte cell line: stimulation of cyclic AMP formation by CGRP and VIP/PHM, and inhibition by NPY through G protein-coupled receptors. AB - Many neuropeptides are present in the peripheral nerves of human skin and are distributed from the intraepidermis to subcutaneous appendages, and those peptides are considered to be involved in the pathogenesis of various inflammatory dermatoses. In this investigation, we determined the effects of various neuropeptides on intracellular cyclic adenosine-5'-monophosphate (AMP) formation in cultured human keratinocytes. Among the many peptides tested, calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), peptide histidine-methionine (PHM), and growth hormone releasing factor (GRF) stimulated a rapid and marked formation of intracellular cyclic AMP in keratinocytes in a dose-dependent manner. The direct association of the receptors for CGRP and VIP with adenylyl cyclase in keratinocytes was confirmed by the findings that CGRP and VIP stimulated the enzyme activity in membrane preparations derived from cultured keratinocytes in the presence of guanosine triphosphate (GTP). On the other hand, neuropeptide Y (NPY) showed an inhibitory effect on forskolin-induced cyclic AMP accumulation in keratinocytes. This inhibitory effect of NPY was completely eliminated by glucocorticoid pretreatment of cultured keratinocytes. Furthermore, the presence of peptides that substantially increase intracellular cyclic AMP accumulation also stimulated DNA synthesis and proliferation in a human keratinocyte cell line in a dose-dependent manner. These results suggest that neuropeptides work directly as biologic modulators of keratinocytes through the cyclic AMP cascade. PMID- 8228324 TI - Upper human hair follicle contains a subpopulation of keratinocytes with superior in vitro proliferative potential. AB - We and others have shown previously that corneal keratinocyte stem cells can proliferate in vitro better than their progeny cells. In this paper, we applied this approach to the identification of hair follicular stem cells. When human scalp hair follicles were placed in explant culture, the bulge area yielded best outgrowths. In another experiment, we isolated different subpopulations of human follicular keratinocytes by micro-dissection, dispersed them by trypsin/EDTA into single cells, and grew them in the presence of 3T3 feeder cells. The keratinocytes were then subcultured under identical conditions to compare their in vitro life span. Our results indicate that the life span of keratinocytes of the upper follicle (containing mainly the isthmus area) > sebaceous gland > lower follicle (between the bulge and bulb) > bulb (containing the matrix cells). The cultured upper follicular keratinocytes tend to be small and relatively uniform in size. The poor in vitro growth of matrix cells may reflect their non-stem cell nature and/or special growth requirement(s) satisfied in vivo by the neighboring dermal papilla cells. Unexpectedly, we found that the upper follicular keratinocytes grow even better than epidermal keratinocytes. The existence of a subpopulation of keratinocytes with an in vitro growth potential superior than other known keratinocytes of the skin supports the hypothesis that follicular stem cells reside in the upper follicle. Our data also raise the possibility that putative follicular stem cells are involved not only in forming the follicle, but also in the long-term maintenance of the epidermis. Finally, we discuss the possibility that keratinocyte stem cells, as defined by their in vivo slow cycling nature, are absent in culture. PMID- 8228325 TI - Identification and characterization of two isozymic forms of arylamine N acetyltransferase in Syrian hamster skin. AB - Arylamine N-acetyltransferase (EC 2.3.1.5) activity was examined using skin from Syrian hamster. Two isozymes of arylamine N-acetyltransferase, designated NAT-1 and NAT-2, were detected on anion-exchange high-performance liquid chromatography analysis. Both enzyme activities had indistinguishable molecular masses (30 kDa), but differed significantly in their specificity toward the aromatic amines including serotonin, dopamine, methoxytryptamine, tryptamine, para-phenetidine, para-aminobenzoic acid, and sulphamethazine. Specifically, NAT-2 but not NAT-1 catalyzed acetylation of dopamine to N-acetyldopamine and acetylation of serotonin to form N-acetylserotonin, a direct precursor of melatonin. The two isozymes were also distinguishable based upon their sensitivity toward methotrexate inhibition (50% inhibiting dose for NAT-1 = 380 microM; NAT-2 > 2 mM). The presence of these two activities in the skin raises new questions about the physiologic role of this enzyme in general and in the skin-specific functions in particular. PMID- 8228326 TI - Treatment of human melanocytes and S91 melanoma cells with the DNA repair enzyme T4 endonuclease V enhances melanogenesis after ultraviolet irradiation. AB - Tanning is a protective response of ultraviolet (UV)-irradiated skin that decreases damage from subsequent sun exposures by increasing the epidermal content of melanin, a brown-black pigment that absorbs light energy throughout the UV and visible portions of the electromagnetic spectrum. The melanin pigment is made by epidermal melanocytes and transferred to surrounding keratinocytes. The action spectrum, time course, and histologic features of tanning are well studied, but the initiating molecular events are unknown. Previous work has shown that T4 endonuclease V, a prokaryotic DNA repair enzyme that catalyzes the first and rate-limiting step in repair of UV-induced pyrimidine dimers, delivered in carrier liposomes (T4N5), enhances repair of UV-induced DNA damage in cultured human cells and protects against photocarcinogenesis in an animal model. We now report that T4N5 treatment enhances UV-induced melanogenesis, as measured by melanin content, tyrosinase activity, 14C-dopa incorporation, and visual assessment in both S91 murine melanoma cells and human melanocytes. T4N5 treatment also increases cell yields following UV irradiation. These data suggest that tanning can be stimulated through enhanced DNA repair. PMID- 8228327 TI - Inhibitory effect of melanin pigment on sensitization and elicitation of murine contact photosensitivity: mechanism of low responsiveness in C57BL/10 background mice. AB - We have shown that murine contact photosensitivity (CPS) to 3,3',4',5 tetrachlorosalicylanilide (TCSA) is genetically controlled mainly by the major histocompatibility complex. The H-2b,d haplotypes are closely associated with high responders, whereas mice with the H-2k are non-responders. Irrespective of their H-2 haplotypes, the C57BL/10 (B10) background strains, including B10, B10.D2, B10.A, and B10.BR, possessing black fur color, were low or nonresponders in CPS to TCSA. In B10 mice, however, high-sensitivity responses were induced when subcutaneous inoculation of epidermal cells (ECs) photomodified in vitro with TCSA was used for both immunization and challenge, suggesting that the epicutaneous route for induction and elicitation is defective in B10 background mice. F1 mice obtained by crossing high-responder BALB/c and low-responder B10 mice, possessing agouti fur color, were non-responders of CPS. The magnitude of CPS in the F2 mice derived from F1 (BALB/c X B10) siblings varied from low to high. When these F2 mice were divided into five groups with regard to fur color, the magnitude of reaction was correlated with the fur color and there was inverse relationship between the magnitude of CPS and the amount of melanin pigment in earlobe ECs. Furthermore, the in vivo formation of TCSA-EC photoadducts was negatively correlated to the melanin amount in earlobes. These observations suggested that the failure in CPS of the B10 background mice stems from inability of in vivo photocoupling of TCSA to ECs, presumably due to absorption of ultraviolet radiation by melanin pigment. PMID- 8228328 TI - Novel and known protein tyrosine kinases and their abnormal expression in human melanoma. AB - We have used the polymerase chain reaction and Northern blotting to identify protein tyrosine kinases that may play an important role in the process of melanoma initiation and progression. Degenerate primers from the conserved catalytic domain of tyrosine kinase genes were used to amplify and clone partial cDNA sequences from a human melanoma cell line (DX3-LT5.1) and normal human melanocytes. When the melanoma reaction products were sequenced, 13 distinct clones were found, of which one is novel to date and has provisionally been named MEK (for melanocytic kinase). Of the remaining 12 known kinases, only two, ERB-B2 and IGF1-R, have previously been reported in pigment cells. Reaction products from melanocytes included only eight of these 13 sequences. To test for quantitative differences in tyrosine kinase expression between normal and malignant cells, a panel of eight melanoma lines and normal melanocytes was analyzed by Northern blotting. Two tyrosine kinases (JTK-14/TIE and TYRO-9) were detected in some melanomas but were not found in normal melanocytes, whereas others, including MEK, appeared to be overexpressed in some malignant lines. A minority of kinases showed either no change or a reduction in the level of mRNA. Expression of tyrosine kinases varied independently, and individual lines contained various combinations of these enzymes. Our findings are consistent with an increased overall expression of these putative growth factor receptors during melanoma development. PMID- 8228329 TI - Melanin reduces ultraviolet-induced DNA damage formation and killing rate in cultured human melanoma cells. AB - Epidermal melanin pigment is believed to prevent development of ultraviolet (UV) induced skin cancer by shielding cell nuclei and reducing DNA damage formation. It has not been experimentally proved, however, whether melanin reduces UV induced DNA damage, because published experiments have been inconclusive. The present study was carried out to determine whether intracellular melanin protected cultured cells against UV-induced DNA damage and killing. Three human melanoma cell lines containing different amounts of melanin were used. Absorption spectrum, subcellular localization of melanin, and melanin concentration were examined in the three cell lines. Two types of DNA damage, cyclobutane pyrimidine dimers and (6-4)photoproducts, were detected by an enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies specific for these photolesions. We found that melanin reduced the induction rates of both types of DNA damage in pigmented cells irradiated with low doses of UV in a melanin concentration dependent manner. Almost no differences in repair capacity for the two types of photolesions were observed among the three melanoma cell lines. We also found that the more highly melanotic melanoma cell lines were more UV resistant than the less melanotic melanoma cell lines. These results suggest that intracellular melanin plays an important role in preventing UV-induced cell killing by reducing the formation of two types of DNA damage. PMID- 8228330 TI - IL-8 gene expression and production in human keratinocytes and their modulation by UVB. AB - Interleukin (IL)-8 is a member of the supergene family of proinflammatory and chemotactic cytokines recently termed chemokines. IL-8 has been implicated in the pathogenesis of inflammatory skin diseases such as psoriasis. In this study, IL-8 mRNA expression and protein production were determined in normal cultured human epidermal keratinocytes after ultraviolet-B (UVB) irradiation. Messenger RNA levels were determined by the reverse transcriptase-polymerase chain reaction (RT PCR) method. Total RNA was extracted from cultured keratinocytes at various time points post-irradiation, reverse transcribed to cDNA, and amplified by PCR using a labeled specific primer for the target gene. Amplified products were sized by electrophoresis, visualized by autoradiography, and quantitated by densitometry. Autoradiographs were normalized relative to glyceraldehyde-3-phosphate dehydrogenase (G3PDH) signals. Constitutive expression of IL-8 mRNA was seen in normal cultured keratinocytes. After 100 or 300 J/m2 UVB irradiation, a rapid increase in IL-8 mRNA level was observed within 1 h after irradiation. At 24 h after irradiation, the mRNA level was elevated 11-13 times compared with the control level. Production of IL-8 protein in culture supernatants was assayed by enzyme-linked immunosorbent assay (ELISA). Significant levels of IL-8 protein were observed at 24 h after irradiation. Cycloheximide treatment blocked this IL 8 protein induction. As IL-8 is known to be an inflammatory cell chemotactic factor, these results suggest a possible role for IL-8 in UVB-induced skin inflammation and diseases. PMID- 8228331 TI - T-lymphocyte clones initiated from lesional psoriatic skin release growth factors that induce keratinocyte proliferation. AB - To investigate whether growth factors derived from T cells in psoriatic lesions are able to stimulate keratinocyte growth, T-cell lines were initiated from lesional psoriasis skin and cloned by limiting dilution. Eight clones with good proliferative capacity out of 40 clones from one patient were stimulated. After 24 h, the conditioned medium was harvested and the growth modulatory effect of the conditioned medium on keratinocytes was assessed. Seven of the eight T-cell clones stimulated keratinocyte growth to an extent ranging from 22% +/- 19 to 64% +/- 9 (mean +/- SD of three experiments) of maximal inducible keratinocyte growth, and one T-cell clone had no effect (-5% +/- 2) on keratinocyte growth. Keratinocyte growth was also induced by T-cell clones obtained from two other patients. Several cytokines were tested in this system to determine which T-cell growth factor may induce the keratinocyte growth. None of the cytokines interferon-g, transforming growth factor-beta, interleukin (IL)-2, IL-3, IL-4, IL 6, IL-8, or granulocyte-macrophage colony stimulating factor alone was found to possibly be responsible for the T-cell-induced keratinocyte growth. Thus the nature of the T-cell keratinocyte growth-promoting stimulus remains to be elucidated. PMID- 8228332 TI - Vitamin D3 and calcipotriol decrease extracellular plasminogen activator activity in cultured keratinocytes. AB - Vitamin D3, 1 alpha,25(OH)2D3, and its metabolites regulate the growth and differentiation of several cell types. Vitamin D3 and its analogue, calcipotriol (MC 903), inhibit the proliferation of cultured human and mouse keratinocytes and induce keratinocyte differentiation. Calcipotriol is effective in the treatment of psoriasis in which increased plasminogen activator activity has been reported. We analyzed therefore the effects of calcipotriol and vitamin D3 on the production of plasminogen activator (PA) activity in human keratinocytes and a mouse keratinocyte cell line. Caseinolysis-in-agarose assays indicated that vitamin D3 decreases total PA activity in both keratinocyte culture systems. Zymographic analyses of the medium indicated that the secreted activator was of the urokinase type (u-PA). A decrease was observed also in extracellular matrix and membrane-associated u-PA activity of vitamin D3 and calcipotriol treated cells. Immunoblotting analysis of the conditioned medium from human keratinocytes revealed a decrease in the u-PA protein levels. Accordingly, Northern hybridization analysis of the respective mRNAs indicated a rapid decrease in urokinase mRNA levels. Calcipotriol decreased u-PA activity also in the presence of inducers of u-PA activity like transforming growth factor-beta, epidermal growth factor, and phorbol-12-myristate-13-acetate. Calcipotriol also caused a decrease in tissue type PA (t-PA) activity of the keratinocytes. Most t-PA activity was associated with the extracellular matrices and cell membranes as revealed by zymographic analysis. Paradoxically, the secretion and deposition of the matrix of plasminogen activator inhibitor type 1 decreased in calcipotriol treated cells. The results indicate that a major effect of vitamin D3 on cultured keratinocytes is a decrease of plasminogen activator activity. PMID- 8228333 TI - Regulation of keratinocyte growth, differentiation, and vitamin D metabolism by analogs of 1,25-dihydroxyvitamin D. AB - 1,25(OH)2D has numerous actions on many tissues. Analogs of 1,25(OH)2D are being sought that are selective, to further an understanding of the mechanisms of action of 1,25(OH)2D and to improve its therapeutic efficacy. Toward these ends we examined eight analogs of 1,25(OH)2D for their ability to regulate 25OHD metabolism by keratinocytes. Choosing the three most potent, we then examined their ability to inhibit keratinocyte proliferation, stimulate cornified envelope formation (a marker of differentiation), and bind to the 1,25(OH)2D receptor (VDR). 1,25(OH)2-24F2-D, 1,25(OH)2-delta 16-D, and 1,25(OH)2-delta 16,23yne-D proved the most potent in inhibiting 1,25(OH)2D production and stimulating 24,25(OH)2D production, being approximately 10-100 times more potent than 1,25(OH)2D itself. 1,25(OH)2-delta 16-D had the highest affinity for the VDR (fourfold higher than that for 1,25(OH)2D itself) and had the greatest ability both to inhibit proliferation and to stimulate differentiation. 1,25(OH)2-delta 16,23yne-D also had a higher affinity for the VDR but was of less or equal potency in stimulating cornified envelope formation and inhibiting proliferation. 1,25(OH)2-24F2-D, which was the most potent regulator of 25OHD metabolism, had a lower affinity for the VDR and was less potent than 1,25(OH)2D in inhibiting proliferation. Our results indicate that even in the same cell different analogs have different rank orders of potency for the various actions of 1,25(OH)2D. PMID- 8228334 TI - Keratinocyte differentiation is associated with changes in the expression and regulation of phospholipase C isoenzymes. AB - In murine keratinocytes, Ca(++)-induced terminal differentiation is accompanied by a rapid and sustained increase of inositol phosphates and diacylglycerol. Based on Western blotting analysis, basal keratinocytes cultured in 0.05 mM Ca++ medium express phospholipase C (PLC)-gamma 1 predominantly and no detectable PLC beta 1. Differentiating keratinocytes cultured in 1.4 mM Ca++ express two- to threefold more PLC-gamma 1 protein and PLC-delta 1, but no detectable PLC-beta 1. Although the amount of PLC-gamma 1 and -delta 1 protein increased, PLC-gamma 1 and -delta 1 mRNA decreased in differentiating cells. Thus the sustained rise of PLC activity induced by Ca++ in differentiating keratinocytes may be associated with higher amounts of both PLC-gamma 1 and -delta 1 in maturing cells, determined by a posttranscriptional mechanism. Tyrosine phosphate content in PLC gamma 1 was low in basal cells and did not change in cells exposed to 1.4 mM Ca++. However, genistein inhibited the increase in PLC activity induced by 1.4 mM Ca++. In contrast, transforming growth factor (TGF)alpha, which stimulates both PLC activity and growth in basal keratinocytes, increased tyrosine phosphorylation of PLC-gamma 1. These results suggest that tyrosine phosphorylation of PLC-gamma 1 by the epidermal growth factor (EGF) receptor is linked to stimulated proliferation, whereas stimulation of PLC activity by Ca++ is linked to keratinocyte differentiation and involves the action of a tyrosine kinase but not tyrosine phosphorylation of PLC-gamma 1. Based on studies using the intracellular free Ca++ chelator BAPTA, a rise in intracellular free Ca++ was not required for stimulation of PLC activity by raising extracellular Ca++. Phorbol esters inhibited PLC stimulation by 1.4 mM Ca++ medium and increased serine phosphorylation of PLC-gamma 1. Exogenous phosphatidylinositol-specific and phosphatidylcholine-specific bacterial PLC also inhibited endogenous inositol phosphate formation and increased endogenous diacylglycerol (DAG). Thus, direct serine phosphorylation of PLC-gamma 1 by protein kinase C is associated with the inhibition of Ca(++)-mediated PLC stimulation. These results show that keratinocytes have multiple mechanisms to regulate PLC activity in response to a specific signal. PMID- 8228335 TI - Characteristics of oligonucleotide uptake in human keratinocyte cultures. AB - Oligodeoxyribonucleotides have the potential to interfere selectively with cellular protein synthesis by sequence-specific hybridization to DNA or RNA molecules. We have investigated the properties of uptake and intracellular localization of fluorescently labeled oligonucleotides in cultured human keratinocytes using confocal laser scanning microscopy. Unlike many other cell types studied, keratinocytes can internalize oligonucleotides without apparent sequestration in endosomes or cell surface accumulation. Uptake is primarily nuclear and unaltered by sodium azide, monensin, or chloroquine pretreatment. We have verified our results with two different fluorophores, fluorescein and Bodipy, and found similar uptake and distribution patterns in both live and fixed cell populations. Surprisingly, we have found uptake to be heterogeneous within a population, with 15-30% of cells internalizing the oligonucleotides. This percentage is drastically increased to roughly 80% at cell population margins, and after release from M phase arrest. These results on uptake and intracellular localization suggest that keratinocytes may have increased sensitivity as target cells for oligonucleotide based gene regulation strategies. PMID- 8228336 TI - Annexin-1 localization in human skin: possible association with cytoskeletal elements in keratinocytes of the stratum spinosum. AB - Annexin-1 (also called lipocortin-1 or p35), a putative substrate of the epidermal growth factor/receptor kinase, protein kinase C, and transglutaminase, was immunolocalized in embryonic, neonatal, adult, and diseased human epidermis. In embryonic skin intense annexin-1 immunoreactivity was found in the periderm at 54 d estimated gestational age (EGA). Later (EGA = 91-143 d), annexin-1 immunoreactivity was restricted to basal keratinocytes. In neonatal skin, basal cells were often more heavily stained than were suprabasal keratinocytes, which were also stained. Only basal keratinocytes stained in adult plantar skin, but in thin skin annexin-1 was present in the basal, suprabasal, and sometimes even in the granular layers of the epidermis. Often, annexin-1 appeared concentrated around the perimeter of cells, especially tonofilament/desmosome-rich keratinocytes of the spinous-cell layer. At high magnification, annexin-1 appeared associated with distinct structures and was very granular in appearance in the intensely stained ductal keratinocytes of eccrine sweat glands, cells that are very highly enriched in keratin tonofilaments. This striking distribution in certain keratinocytes enriched in tonofilaments suggests a role for annexin-1 in cytoskeletal functions. PMID- 8228337 TI - Nicein (BM-600) in junctional epidermolysis bullosa: polyclonal antibodies provide new clues for pathogenic role. AB - We have raised polyclonal antibodies against each of three subunits of the new basement membrane component nicein (formerly BM-600), the antigen recognized by the monoclonal antibody GB3 (Biochem Biophys Acta 942:45-56, 1988). Preparation of such antibodies was achieved from gel electrophoresis purification of nicein isolated by immuno-affinity chromatography. These antibodies were reactive with each transblotted denatured nicein subunit and recognized the native protein both in cultured keratinocytes and in all normal human basement membranes where the GB3 antigen is located. A reciprocal immuno-cross-reactivity was detected with the antibodies directed against the 100-kD and 150-kD (sometimes resolved as a 146-150-kD doublet) subunits of nicein, showing that they share some identical epitopes. In tissues and keratinocyte cultures from patients with the Herlitz form of junctional epidermolysis bullosa (H-JEB), GB3 is unable to recognize nicein, and the question arises whether this is due to an absence of synthesis or a structural abnormality of the protein. We report here that the polyclonal antibody directed against the 150-kD subunit of nicein binds its antigen in H-JEB patients (although usually less intensely than in control skin), whereas the other two antibodies either do not recognize or recognize only weakly their respective antigen subunits. These data suggest that nicein is present but structurally altered in basement membranes from H-JEB tissues. Furthermore, in non-Herlitz junctional and dystrophic types of epidermolysis bullosa, all three polyclonal antibodies recognize their antigens normally. Consequently, such antibodies should serve as potentially useful molecular tools for studying the expression of nicein in H-JEB. PMID- 8228338 TI - Ultraviolet hypermutability of a shuttle vector propagated in xeroderma pigmentosum variant cells. AB - Patients with the variant form of xeroderma pigmentosum (XP) have clinical XP including a high frequency of skin cancer but, in contrast to the other forms of XP, have normal post-ultraviolet (UV) DNA excision repair and nearly normal post UV survival. However, like excision repair-deficient XP cells, the XP variant cells are UV hypermutable. We used a UV-treated plasmid shuttle vector, pZ189, to examine the DNA repair defect in lymphoblastoid cells from an XP variant patient, XPPHBE, and a normal control. Plasmid repair, mutagenesis, and replication occur within transfected cells in a process dependent on the cells' repair capacity. With the XP variant cells post-UV, plasmid survival was normal with but there was an abnormally increased post-UV plasmid mutation frequency. Sequence analysis of the mutated plasmids revealed an increased frequency of plasmids with single base substitution mutations with the XP variant cells. As in earlier studies with UV mutagenesis, there was a predominance of G:C-->A:T base substitution mutations with plasmids recovered from both cell lines. The frequency of G:C-->C:G transversions was significantly higher with plasmids recovered from the XP variant cells than from normal cells. The location of mutations in the marker gene was non-random with different mutagenic hotspots found in plasmids recovered from the XP variant cells and from the normal cells. This study suggests that plasmid UV hypermutability in the presence of normal UV survival may be related to the increased UV skin cancer susceptibility of XP variant patients. PMID- 8228339 TI - Transepidermal induction of contact hypersensitivity in mice with a water-soluble hapten. AB - An experimental analysis has been conducted on the capacity of a highly-reactive, water soluble hapten, trinitrobenzene sulfonic acid, to induce contact hypersensitivity when applied epicutaneously to body wall skin of normal mice. Application of plastic chambers containing the hapten to murine skin for as little as 1 h produced readily detectable sensitization in several genetically disparate inbred strains. Moreover, the efficiency of sensitization was found to be similar to that following epicutaneous application of this hapten's lipid soluble cogener, trinitrochlorobenzene. Using this approach, it has been determined that UVB radiation, intradermally injected tumor necrosis factor alpha, and epicutaneously applied cis-urocanic acid can impair contact hypersensitivity induction by transepidermally delivered trinitrobenzene sulfonic acid, and that animals that fail to become sensitized proceed to acquire hapten specific unresponsiveness. It is concluded that epicutaneous sensitization to chemically reactive, water-soluble molecules is experimentally attainable if precautions are taken to insure that contact between the hapten solution and the cutaneous surface is maintained for at least 1 h. Understanding the cellular and molecular mechanisms of sensitization and tolerance induction by epicutaneously applied water soluble haptens may prove to be important in understanding the pathogenesis of allergic contact dermatitis that develops to chemicals in the industrial setting, in the environment, and in the clinic. PMID- 8228340 TI - Differential mortality by measles vaccine titer and sex. AB - Mortality was evaluated in 1972 children who had received measles vaccines at 6 11 months of age that were 10-fold (medium titer) or 100-fold (high titer) greater than standard titer. Mortality among boys did not differ by vaccine titer and was similar to mortality in children who received standard-titer vaccine. Girl recipients of high-titer vaccine had somewhat greater mortality than girls who received medium-titer vaccine (risk ratio = 1.71, 95% confidence interval = 0.91-3.24). Increased mortality was associated with high-titer vaccine for girls but not for boys (P = .04). There was no evidence of selection bias or preferential health care by sex that might explain the differential mortality. This mortality pattern has been noted in two other populations with high background infant and childhood mortality. The biologic basis for this effect on mortality has not been determined. Data from this and other studies have resulted in discontinuation of the use of high-titer measles vaccines. PMID- 8228341 TI - Immunologic parameters 2 years after high-titer measles immunization in Peruvian children. AB - Immunization with high-titer measles vaccines has been associated with excess mortality in children 2-4 years after vaccination. In this study, immunologic parameters in 64 Peruvian children who had been immunized an average of 27 months earlier with high-titer vaccines were compared with parameters in 76 recipients of low-titer vaccines. Delayed-type hypersensitivity, lymphocyte phenotype distributions by flow cytometry, and lymphoproliferation after phytohemagglutinin (PHA) stimulation were assessed. High-titer recipients had smaller indurations to tetanus, diphtheria, and Proteus (P < .05) antigens, decreased PHA stimulation (P = .04), and a lower percentage of CD4+ lymphocytes (P = .04) than low-titer recipients. After adjustment for sex, concurrent illnesses, and other variables in regression analyses, high-titer recipients had a lower percentage of CD4+ lymphocytes (P = .025) and decreased lymphocyte proliferation to PHA (P = .058). These results may provide a clue to the pathogenesis of delayed excess mortality after high-titer measles vaccination in some developing countries. PMID- 8228342 TI - Shedding of virulent poliovirus revertants during immunization with oral poliovirus vaccine after prior immunization with inactivated polio vaccine. AB - Fecal shedding of virulent revertant polioviruses was examined in isolates from infants previously immunized with > or = 1 dose of orally administered live attenuated polio vaccine (OPV) alone, enhanced-potency inactivated polio vaccine (EIPV) alone, or a combination of both. After administration of OPV alone, vaccine poliovirus serotypes were recovered in feces within 1 week and for as long as 31-60 days in 30%-80% of subjects after 1 or 2 doses and in 30%-50% after immunization with > or = 3 doses. No revertant poliovirus shedding was observed after OPV challenge in subjects immunized previously with > or = 3 doses of OPV. However, fecal shedding of revertant poliovirus after OPV challenge was observed in 50%-100% of subjects previously immunized with > or = 3 doses of the EIPV. These findings suggest that prior immunization with EIPV does not prevent fecal shedding of revertant polioviruses after subsequent reexposure to OPV. PMID- 8228343 TI - Ultrastructural analysis of circulating cytomegalic cells in patients with active cytomegalovirus infection: evidence for virus production and endothelial origin. AB - The presence of cytomegalic inclusion cells in the peripheral blood of patients with an active cytomegalovirus infection has recently been demonstrated. Immunologic staining showed that these cells were of endothelial origin. Study of circulating cytomegalic cells by transmission electron microscopy showed the cells to be productively infected with cytomegalovirus. Viral capsids were present in the nucleus and virus particles and dense bodies were found in the cytoplasm. The results indicate that these circulating cytomegalic cells could disseminate cytomegalovirus throughout the body. In addition, the finding of a cluster of cytomegalic cells in the peripheral blood linked together by zonula adherens type cell junctions is further evidence that these cells are of endothelial origin and suggests that the endothelial damage may be extensive. PMID- 8228344 TI - Antibodies induced by a primary cytomegalovirus infection react with human herpesvirus 6 proteins. AB - After a primary human cytomegalovirus (HCMV) infection, antibody titer to human herpes-virus 6 (HHV-6) rises. To determine if this occurs because of simultaneous infection with both viruses, serologic responses to these viruses were investigated among healthy women who received a live HCMV vaccine or acquired HCMV from an infected child. Both vaccination and natural infection caused four fold or greater titer rises to HHV-6. Analysis of sera from 5 children with primary HHV-6 showed no serologic response to HCMV. Absorption of 7 sera with HCMV antigens reduced titers to HHV-6 by two- to fourfold. Immunoblot analysis of sera obtained from 10 persons before and after a primary HCMV infection revealed that after HCMV infection, 2 persons developed IgG reactivity to the 116-kDa protein (gp116) of HHV-6 and to two cleavage products of this protein. Reactivity to HHV-6 gp116 appeared after HCMV sero-conversion and was removed by absorption of sera with HCMV glycoprotein gB (gB), indicating that antibodies to HCMV gB cross-react with HHV-6 gp116, the likely gB homologue of HHV-6. The serologic diagnosis of HHV-6 infection requires excluding a primary HCMV infection. PMID- 8228345 TI - Direct contact with herpes simplex virus-infected cells results in inhibition of lymphokine-activated killer cells because of cell-to-cell spread of virus. AB - Natural killer (NK) and lymphokine-activated killer (LAK) cells are disarmed after contact with herpes simplex virus (HSV)-infected cells. Cells infected with HSV-1 mutants that lack glycoproteins essential for viral entry into cells (gB, gD, gK, gH, and gL) did not inhibit LAK cells; cells infected with HSV-1 mutants that lack glycoproteins not required for virus entry into cells (gE, gI, gG, and gJ) inhibited lysis. LAK cells became infected after contact with target cells infected with wild-type HSV-1 but not with a gD-HSV-1, which cannot spread from cell to cell. Because LAK cells were inhibited only by very high concentrations of cell-free preparations of HSV and because neutralizing antibodies did not prevent infection of LAK cells in contact with infected cells, infection of LAK cells is probably greatly enhanced by the apposition of the effector and target cell membranes during target recognition. Disarming of immune effector cells by infection may be a general strategy for immune evasion by HSV. PMID- 8228346 TI - Low frequency of cytotoxic liver-infiltrating T lymphocytes specific for endogenous processed surface and core proteins in chronic hepatitis B. AB - To investigate the role of hepatitis B virus (HBV)-specific CD8+ T cells in chronic hepatitis B, the lytic activity of peripheral blood mononuclear cells (PBMC) and liver-infiltrating T cell clones and cytotoxic T cell (CTL) lines stimulated by recombinant vaccinia virus-infected cells were analyzed. Autologous and allogeneic Epstein-Barr virus-transformed B cells infected with vaccinia vectors (VAC) that contain sequences of the surface (S), secretory core (E), cytoplasmatic core (C) VAC antigen of HBV, or the wild-type (WT) VAC served as target cells. ELISA and immunoblotting showed HBV antigen expression in infected cells. Neither PBMC nor C- or E-VAC-stimulated CTL lines showed specific lytic activity. However, S-VAC stimulated blood- and liver-derived CTL from 2 patients with hepatitis B lysed autologous S-VAC but not WT-VAC-infected target cells. One of 158 CD8+ T cell clones from 6 patients with active hepatitis B lysed autologous, but not allogeneic, S-VAC-infected targets. Thus, CTL are present among liver-infiltrating T cells in chronic hepatitis B that recognize endogenously processed hepatitis B surface antigen but not HBc or HBe antigen. The lack of core-specific CTL may contribute to failure in virus elimination in chronic inflammation. PMID- 8228347 TI - Lack of T cell dysfunction and programmed cell death in human immunodeficiency virus type 1-infected chimpanzees correlates with absence of monocytotropic variants. AB - In asymptomatic human immunodeficiency virus (HIV) infection in humans, disturbed T cell functions such as anergy and programmed cell death, thought to result from inappropriate signaling by antigen-presenting cells due to HIV infection, precede increase in virus load, decline in CD4+ T cell numbers, and subsequent disease progression. Here, in 3 long-term HIV-1-infected asymptomatic chimpanzees, antigen-presenting cell function was intact and T cells had normal proliferative capacity with no evidence of HIV-1-associated programmed cell death. Polymerase chain reaction analysis demonstrated low frequencies of cells harboring proviral DNA. Primary virus isolation from the infected animals demonstrated the absence of monocytotropic HIV-1 variants, in concordance with complete insusceptibility of chimpanzee monocytes for HIV-1 infection. Possibly, because of the incapacity of HIV-1 to infect monocytes, systemic immune dysfunction will not occur, contributing to controlled viral replication and maintenance of the asymptomatic state in HIV-infected chimpanzees. PMID- 8228348 TI - Inhibition of the protease of human immunodeficiency virus blocks replication and infectivity of the virus in chronically infected macrophages. AB - Because of the importance of monocytes/macrophages (M/M) as an in vivo reservoir of human immunodeficiency virus (HIV), a study was done to investigate whether viral replication in chronically infected macrophages (HIV M/M) could be inhibited by various drugs, including U-75875, an inhibitor of HIV protease. HIV replication in M/M and in chronically infected T cells was dramatically decreased by U-75875, while other drugs, including zidovudine, interferon-alpha, and an antisense oligodeoxynucleotide against the rev gene, were effective antiviral agents only in de novo-infected cells. Virus titer in HIV M/M was reduced approximately 10(5)-fold by nontoxic concentrations of U-75875, while no effect on HIV DNA or virus antigen expression on cell membrane was achieved in M/M infected either chronically or de novo. Thus, U-75875 essentially worked against late stages of viral replication. These data support the use of protease inhibitors, alone or in combination, in the therapy of HIV-infected patients. PMID- 8228349 TI - Human immunodeficiency virus type 1 replication in peripheral blood mononuclear cells in the presence of cocaine. AB - Previous studies have shown that cocaine consumption alters the normal immune functions of animals. Several epidemiologic studies have indicated an association between cocaine consumption and an increased risk for AIDS. In the present studies, unstimulated peripheral blood mononuclear cells (PBMC) isolated from 8 healthy, human immunodeficiency virus (HIV) type 1-seronegative volunteers were exposed to cocaine or one of its by-products, in vitro, at concentrations compatible with blood levels found during clinical abuse of cocaine. PBMC treated with cocaine had significantly increased levels of HIV-1 replication after infection in vitro compared with untreated PBMC. The major cocaine by-product, benzoylecgonine, did not appear to exert any significant potentiating effect on HIV-1 replication. Cocaine or its by-product did not significantly increase HIV-1 replication in persistently HIV-1-infected T lymphocytic or monocytoid cell lines. These data indicate that exposure of PBMC but not chronically HIV-1 infected cell lines to cocaine can result in increased HIV-1 replication. PMID- 8228350 TI - Safety, immunogenicity, and transmissibility of single-dose live oral cholera vaccine strain CVD 103-HgR in 24- to 59-month-old Indonesian children. AB - Recombinant A-B+ Vibrio cholerae O1 strain CVD 103-HgR is a safe, highly immunogenic, single-dose live oral vaccine in adults in industrialized countries. Safety, excretion, immunogenicity, vaccine transmissibility, and environmental introduction of CVD 103-HgR were investigated among 24- to 59-month-old children in Jakarta. In 81 households, 1 child was randomly allocated a single dose of vaccine (5 x 10(9) cfu) and another, placebo. Additionally, 139 unpaired children were randomly allocated vaccine or placebo. During 9 days of follow-up, diarrhea or vomiting did not occur more often among vaccines than controls. Vaccine was minimally excreted and was isolated from no controls and from 1 (0.6%) of 177 unvaccinated family contacts. A 4-fold or higher rise in serum vibriocidal antibody was observed in 75% of vaccines (10-fold rise in geometric mean titer over baseline). Of 135 paired placebo recipients or household contacts, 5 had vibriocidal seroconversions. Moore swabs placed in sewers and latrines near 97 households failed to detect vaccine. These observations pave the way for a large scale field trial of efficacy. PMID- 8228351 TI - Evaluation of an intervention program for the control of an outbreak of shigellosis among institutionalized persons. AB - After control measures were initiated to stop an outbreak of shigellosis in an institution for the developmentally disabled, there was a sharp decline in the number of cases of Shigella sonnei infection. Among ill residents, those treated with antibiotics had shorter mean duration of diarrhea (2.4 vs. 4.5 days, P < .01) and were less likely to have stool cultures positive for shigellae 2-4 weeks after onset of diarrhea (0/25 vs. 5/19; relative risk [RR] = undefined; P = .02). The attack rate was higher in villages where segregation of ill residents was not practiced (46/73 vs. 53/155; RR = 1.8; 95% confidence limits [CL], 1.4, 2.4). In individual housing units where ill residents were not segregated (preintervention), a correlation was found between mean duration of diarrhea and unit attack rates (r = .88; 95% CL, 0.29, 0.99). A study of all 305 residents 10 weeks after the intervention began revealed no positive stool cultures. PMID- 8228352 TI - Relapse in melioidosis: incidence and risk factors. AB - From 1986 to 1991, 602 patients with melioidosis were seen in Sappasitprasong Hospital, Ubon Ratchatani, Thailand. The in-hospital mortality was 42%. Of 118 adult patients followed long-term, 27 (23%) had culture-proven relapses of melioidosis (3 relapsed twice), a relapse rate of 15% (95% confidence interval [CI], 11-22) per year. The median time from discharge to relapse was 21 weeks (range, 1-290). In 44% of patients, relapses included septicemia, and 27% died. Patients with severe disease (multiple foci of infection or septicemia) relapsed 4.7 times (95% CI, 1.6-14.1) more frequently than patients with localized melioidosis. Underlying disease was not a risk factor, but initial parenteral treatment with ceftazidime reduced the risk of relapse 2-fold (95% CI, 1.1-3.4). Relapses were 3.3 (95% CI, 1.4-9.0) times more frequent following short-course (< or = 8 weeks) oral coamoxiclav than after the oral combination regimen of chloramphenicol, doxycycline, and cotrimoxazole. Longer oral treatment with either reduced relapse 1.6-fold (95% CI, 1.2-1.9). The optimum choice and duration of antibiotic treatment to prevent relapse in melioidosis remain to be determined. PMID- 8228353 TI - A mutation affecting expression of a major outer membrane protein of Moraxella catarrhalis alters serum resistance and survival in vivo. AB - A major outer membrane protein (CopB) of Moraxella catarrhalis is a target for antibodies that enhance clearance of this organism from the lungs of mice. A mini Tn10kan transposon was inserted into the cloned copB gene from M. catarrhalis O35E, and an isogenic mutant unable to express the CopB protein was constructed by transforming this mutated gene into the wild-type strain. The mutant grew at the same rate as the wild-type parent strain in broth. Unlike the serum-resistant parent strain, this mutant was sensitive to killing by normal human serum, and its ability to survive and grow in the lungs of animals was impaired. Genetic restoration of CopB protein expression resulted in the simultaneous acquisition of wild-type levels of serum resistance and the ability to resist pulmonary clearance in vivo. Thus, the CopB protein of M. catarrhalis may be important in the interaction between this organism and the defense mechanisms of the respiratory tract. PMID- 8228354 TI - Interleukin-8 production by human peritoneal mesothelial cells in response to tumor necrosis factor-alpha, interleukin-1, and medium conditioned by macrophages cocultured with Staphylococcus epidermidis. AB - Patients treated with continuous ambulatory peritoneal dialysis (CAPD) may suffer from recurrent peritonitis episodes caused by Staphylococcus epidermidis. Early recruitment of granulocytes from the peripheral blood is important for the peritoneal antibacterial defense of CAPD patients. In this study, human peritoneal mesothelial cells were shown to produce high levels of interleukin-8 (IL-8) in response to IL-1 beta and tumor necrosis factor-alpha (TNF alpha) but not lipopolysaccharide or S. epidermidis. Coculture of peritoneal macrophages with S. epidermidis induced high levels of IL-1 alpha, IL-1 beta, and TNF alpha in 24-h-conditioned medium. Preincubation of this medium with anti-TNF alpha, anti-IL-1 alpha, or anti-IL-1 beta partially blocked stimulation of IL-8 production by mesothelial cells. Added together, these antibodies abolished IL-8 production to a level just above background. Migration of granulocytes to the stimulated mesothelial cell-conditioned medium could be totally blocked with rabbit polyclonal anti-IL-8 antibody. Thus, mesothelial cells are important for the recruitment of granulocytes into the peritoneal cavity. PMID- 8228355 TI - Occurrence of capsular polysaccharide/adhesin among clinical isolates of coagulase-negative staphylococci. AB - Clinical isolates of coagulase-negative staphylococci were analyzed for elaboration of the capsular polysaccharide/adhesion (PS/A) and extracellular biofilm or slime. Of the 151 analyzed, 103 (68%) produced PS/A and 69 (46%) made extracellular slime; 87% of the slime-producing isolates made PS/A. Among isolates from all clinical infections examined except peritonitis, PS/A-positive isolates bound significantly (P < .001) more colony-forming units after 15 min to 1.5-cm segments of silicone-elastomer catheter than did PS/A-negative isolates. Slime-positive isolates were not more adherent than slime-negative isolates, because 42% of the PS/A-positive isolates were slime-negative. Thus, PS/A expression is common among clinical isolates of coagulase-negative staphylococci, accounting for most slime-positive and a proportion of slime-negative isolates. PMID- 8228356 TI - Evidence that Chlamydia pneumoniae causes pneumonia and bronchitis. AB - An epidemic often provides an opportunity to obtain evidence of the etiologic association of a microorganism with disease. Chlamydia pneumoniae TWAR is a newly recognized organism whose relationship to disease is not completely understood. An outbreak of C. pneumoniae infections from November 1990 to February 1991 was studied in University of Washington students. Twelve TWAR infections were identified serologically in 54 students with acute respiratory disease. The organism was isolated from 7 of the 12 and identified by the polymerase chain reaction (PCR) in 2 that were isolation-negative. The organism was not found in any of the 42 serologically negative patients or in 51 control student patients without respiratory illness cultured in 1991. There was no evidence of infection with Mycoplasma pneumoniae or respiratory viruses in the 12 patients with C. pneumoniae infection. During the 4-month outbreak, there was an increase in total pneumonia cases. These findings provide evidence for an etiologic association of C. pneumoniae with pneumonia and bronchitis. PMID- 8228357 TI - Onchocerca volvulus provides ligands for the stimulation of human gamma/delta T lymphocytes expressing V delta 1 chains. AB - Peripheral blood mononuclear cells (PBMC) from 8 onchocerciasis patients, treated or not with ivermectin, were analyzed for phenotypic cell surface markers. A significant increase (P < .05) in gamma/delta T cells expressing the V delta 1 chain compared with normal and endemic controls was detected in all patients. PBMC populations from onchocerciasis patients were not expanded after restimulation with Onchocerca volvulus antigens in vitro, but both V delta 1 and V delta 2 T cells from normal donors were increased significantly in response to O. volvulus and Mycobacterium tuberculosis (P < .05), respectively. Frozen sections of all 5 onchocerca nodules tested demonstrated an increased number of CD3+ cells in the vicinity of the adult worm, in all cases expressing the alpha/beta T cell receptor and in 2 patients also expressing the gamma/delta T cell receptor; 60% of T cells expressed the activation marker Ki67. These data suggest that O. volvulus provides ligands to V delta 1 T cells. PMID- 8228358 TI - Persistent intrathecal immune activation in patients with herpes simplex encephalitis. AB - Neopterin and beta 2-microglobulin (beta 2M) levels were analyzed in sequential cerebrospinal fluid (CSF) and serum samples from 20 patients with herpes simplex encephalitis (HSE) and 30 patients with acute febrile encephalopathy of other cause (non-HSE). Markedly elevated acute phase CSF levels of neopterin and beta 2M were found in 19 HSE patients, but the levels were only moderately increased in most of those with non-HSE. Neopterin levels were analyzed in an additional 15 HSE patients who died within a month of the onset of neurologic symptoms and correlated with the clinical severity of the HSE. After HSE, but not after non HSE, increased levels of neopterin and beta 2M persisted for a long time (> or = 13 years). Specific intrathecal IgG activity persisted in all but 2 HSE patients. These findings indicate that there is a vigorous acute inflammatory response and a long-term persistence of intrathecal cellular and humoral immune activity in HSE. PMID- 8228359 TI - Detection of varicella-zoster virus DNA in nasopharyngeal secretions of immune household contacts of varicella. AB - Persons immune to varicella-zoster virus (VZV) are not at risk for developing clinical infection after exposure to varicella. However, the extent to which they might serve as vectors for the transmission of VZV to others is not known. Information in this regard would be important in establishing hospital infection control policies, especially in relation to the care of immunocompromised hosts. A polymerase chain reaction-based detection system was used to detect the presence of VZV DNA in the nasopharyngeal secretions of household contacts of children with varicella. VZV DNA was identified in 4 of 5 immune adults and 1 susceptible sibling when sampled within 3 days of recognition of a household case of varicella. Further investigations are needed to determine whether this represents a limited window of VZV replication in the nasopharynx of immune persons during which they may serve as vectors of VZV. PMID- 8228360 TI - Inhibition of varicella-zoster virus (VZV) glycoprotein expression by a human monoclonal antibody against VZV glycoprotein III. AB - Effects of human monoclonal antibodies (MAbs) against varicella-zoster virus (VZV) glycoproteins (gp) on VZV glycoprotein expression were analyzed by flow cytometry. Human embryonic fibroblast (HEF) cells were inoculated with cell-free VZV and cultured in the presence of human MAbs. While human MAbs directed against gpI (V2) and -II (V1) did not affect VZV expression, a human MAb against gpIII (V3) inhibited expression of VZV gpI, -II, -III, and -IV. V3 also inhibited spread of VZV from infected to uninfected cells. Interferon-alpha, -beta, and gamma and tumor necrosis factor-alpha were able to synergize with V3 and inhibit VZV gp expression. These results suggest that human MAb V3 may be useful for passive prophylaxis against VZV infection and for treatment of severe VZV infection. PMID- 8228361 TI - Herpes zoster in patients with advanced human immunodeficiency virus infection treated with zidovudine. Zidovudine Epidemiology Study Group. AB - To determine the prevalence, incidence, and effects on disease progression and survival of herpes zoster in patients with advanced human immunodeficiency virus (HIV) disease, data from a multicenter observational cohort study of 1044 patients with AIDS or AIDS-related complex (ARC) and CD4 cell count < or = 0.25 x 10(9)/L treated with zidovudine were analyzed. Of 163 patients (16%) with a history of herpes zoster at enrollment, 22 (13%) had a recurrence during the 2 year follow-up. For those without prior herpes zoster, the probability of its development was 6.3% at 1 and 8.8% at 2 years. Progression to AIDS was not associated with herpes zoster. By proportional hazards analysis, an initial occurrence of herpes zoster was associated with prolonged survival independent of baseline CD4 cell count and disease stage; however, recurrence tended to be associated with death. Thus, herpes zoster is relatively common in advanced HIV infection and its initial occurrence late in disease may indicate improved prognosis. PMID- 8228362 TI - Detection of human T lymphotropic virus type II/b in human immunodeficiency virus type 1-coinfected persons in southeastern Italy. AB - A preliminary screening of 511 persons at risk for AIDS living in southeastern Italy disclosed 20 cases of seroreactivity to human T lymphotropic viruses (HTLV). To verify and type the HTLV infection among these subjects, confirmatory serologic tests, polymerase chain reaction (PCR), and virus culture were done. No evidence of HTLV-I infection was found. HTLV-II infection was confirmed in 8 cases by HTLV-specific, synthetic peptide EIAs and PCR on uncultured cells; restriction analysis of the PCR-amplified env regions revealed the presence of HTLV-II/b strains in all 8 cases. Four sera were nontypeable by EIA. The finding of such indeterminate reactivities in a geographic area in which HTLV variants were previously described indicates the need for more extensive surveys among the healthy population. HTLV-II was isolated in 5 cases, and virus isolation was mostly dependent on the presence of an actively replicating human immunodeficiency virus type 1 in culture. PMID- 8228363 TI - Presence of respiratory syncytial virus genomic sequences in middle ear fluid and its relationship to expression of cytokines and cell adhesion molecules. AB - The presence of respiratory syncytial virus (RSV) and several cytokines and cell adhesion molecules in middle ear effusions and mucosal tissues was evaluated using polymerase chain reaction. RSV genomic sequences were detected in 23 (52.7%) of 44 middle ear effusions tested. The sequences were detectable at an even higher rate (82.4%) in effusions of children in whom infectious virus was detected in the nasopharynx. All samples with the RSV genome contained the mRNA for interleukin-1 beta and -6 and tumor necrosis factor-alpha. The messages for these cytokines, together with intercellular adhesion molecule-1, endothelial leukocyte adhesion molecule-1, and vascular cell adhesion molecule-1, were detected in human middle ear mucosal organ cultures infected in vitro with RSV. Our results suggest that the enhanced synthesis of proinflammatory cytokines and cell adhesion molecules in the middle ear infected with RSV may contribute to the inflammatory processes in otitis media. PMID- 8228364 TI - Comparative trial of large-particle aerosol and nose drop administration of live attenuated influenza vaccines. AB - Healthy adult subjects (n = 198) were randomized to receive bivalent cold-adapted (ca) influenza A vaccine containing 10(7.4) TCID50 each of A/Kawasaki/9/86 (H1N1) and A/Los Angeles/2/87 (H3N2) by either nose drops (ND) or large-particle aerosol (LPA). All subjects had received monovalent inactivated influenza B vaccine intramuscularly in the previous year. Ninety percent of LPA recipients and 82% of ND recipients preferred intranasal administration to their previous experience with intramuscular vaccine. Twenty-six (27%) of 98 LPA recipients and 47 (49%) of 97 ND recipients reported swallowing vaccine (P = .008). Fever was observed uncommonly (< or = 5%), and incidence of respiratory symptoms was comparable between groups. Fourfold or greater hemagglutination antibody response to at least one of the influenza A vaccine viruses was significantly more frequent after LPA (64%) than ND vaccination (43%; P = .005). LPA administration of ca influenza to the nasopharynx was well tolerated, safe, and more immunogenic than ND delivery. PMID- 8228365 TI - Cerebrospinal fluid neopterin concentrations in central nervous system infection. AB - Cerebrospinal fluid (CSF) neopterin levels were determined by RIA in individuals with central nervous system (CNS) or human immunodeficiency virus (HIV) infections and in healthy controls. The mean CSF neopterin concentrations were 63.0 nmol/L in 15 patients with acute bacterial meningitis, 54.9 nmol/L in 15 patients with Lyme neuroborreliosis, 32.5 nmol/L in 10 patients with viral meningitis, 130.9 nmol/L in 8 patients with viral encephalitis, 13.9 nmol/L in 15 patients with asymptomatic HIV infection, 26.0 nmol/L in 11 patients with AIDS without dementia, 65.4 nmol/L in 4 patients with AIDS dementia, and 4.2 nmol/L in 24 healthy controls. Although patients with viral encephalitis had higher mean neopterin levels than any other patient category studied, the CSF neopterin concentrations cannot be used to discriminate between viral and bacterial infections. Analysis of CSF levels of neopterin may be useful as guidance in following clinical course and effect of treatment and can provide information of value in addition to CSF cell count as a measurement of CNS immune stimulation. PMID- 8228366 TI - Efficacy of azithromycin for treating Babesia microti infection in the hamster model. AB - Because of its prevalence and severity, Babesia microti infection is an important public health problem. The current treatment of choice is clindamycin plus quinine. However, in some cases other treatments are needed because of drug intolerance or relapse. The activity of azithromycin was investigated for treatment of babesiosis in the hamster model. All animals received vancomycin to prevent antibiotic-associated colitis. Quinine (250 mg/kg/day), azithromycin (150 mg/kg/day), and the combination of azithromycin and quinine were compared. A significant suppression of parasitemia was found in all treatment groups (combination had the greatest effect, followed by azithromycin, then quinine; P < .05). The mean survival was significantly prolonged in the combination group (P < .05). Azithromycin as monotherapy in a higher dose (300 mg/kg/day) also resulted in a significant prolongation of survival (P < .05). Spirogermanium and ciprofloxacin, which have been reported to have antimalarial activity, had no effect on parasitemia or survival in this experimental babesiosis model. PMID- 8228367 TI - Antibiotic treatment of experimental Pseudomonas aeruginosa pneumonia in guinea pigs: comparison of aerosol and systemic administration. AB - Pseudomonas aeruginosa pneumonia is associated with high mortality despite appropriate systemic antibacterial therapy. To examine the effects of aerosolized tobramycin on experimental P. aeruginosa pneumonia, guinea pigs were assigned to two main groups: a study group, in which P. aeruginosa was instilled into the tracheal lumen, and a control group, in which normal saline was instilled. Each group was then subdivided according to treatment: normal saline or tobramycin by intramuscular injection, normal saline or tobramycin by aerosol, or normal saline or tobramycin via both aerosol and intramuscular injection. Compared with either intramuscular or aerosolized tobramycin, the combination of aerosol plus intramuscular tobramycin achieved slightly higher survival, total eradication of P. aeruginosa from lungs, and a significant further reduction of lung inflammatory changes. Adding aerosolized tobramycin to systemic tobramycin in the treatment of experimental P. aeruginosa pneumonia enhanced eradication of P. aeruginosa and mitigated pulmonary inflammatory changes. PMID- 8228368 TI - Ciprofloxacin and trimethoprim-sulfamethoxazole versus placebo in acute uncomplicated Salmonella enteritis: a double-blind trial. AB - The role of ciprofloxacin and trimethoprim-sulfamethoxazole (TMP-SMZ) was evaluated in empiric treatment of uncomplicated Salmonella enteritis in a comparative, double-blind trial. Patients were randomized to receive ciprofloxacin (500 mg), TMP-SMZ (160/800 mg), or placebo orally twice daily for 5 days. There were 65 evaluatable patients with acute, uncomplicated, culture confirmed Salmonella enteritis. Duration of diarrhea, abdominal pain, or vomiting and time to defervescence were not significantly different for patients treated with ciprofloxacin, TMP-SMZ, or placebo; there also were no significant differences with respect to full resolution of symptoms for ciprofloxacin versus placebo (point estimate, 0.2 days; 95% confidence interval [CI], -0.5 to 0.9 days) or for TMP-SMZ versus placebo (point estimate, 0.2 days; 95% CI, -1.0 to 0.6 days). The rate of clearance of salmonellae from stools was not significantly different among the groups. PMID- 8228369 TI - Protective effect of a recombinant amino-terminal fragment of bactericidal/permeability-increasing protein in experimental endotoxemia. AB - Bactericidal/permeability-increasing protein (BPI), a cationic protein found in neutrophil granules, binds with high affinity to gram-negative bacterial lipopolysaccharide (LPS) and can inhibit its actions in vitro. The in vivo efficacy of a recombinant 23-kDa amino-terminal LPS-binding fragment of BPI (rBPI23) was assessed in a mouse model of lethal endotoxemia. Systemic administration of rBPI23 protected actinomycin D-sensitized mice from lethal LPS (Escherichia coli O111:B4) challenge in a dose-dependent manner, with almost complete protection at the highest dose (10 mg/kg; 93% survival vs. 13% in vehicle-treated controls). Surviving rBPI23-treated animals did not show histopathologic signs of tissue damage evident in control animals that had died after LPS challenge. rBPI23 also attenuated the LPS-induced elevation in serum levels of tumor necrosis factor-alpha and interleukin-1 alpha, mediators believed to be involved in the pathogenesis of endotoxemia and sepsis. Thus, rBPI23 may be a potential new therapeutic agent for the treatment of gram-negative bacterial infection and sepsis. PMID- 8228370 TI - Comparison of the efficacy of polyenes and triazoles against hematogenous Candida krusei infection in neutropenic mice. AB - Candida krusei is reported to cause serious infections in immunocompromised patients, particularly those receiving prophylaxis with antifungal azoles. Treatment of this infection can be very challenging. The efficacy of amphotericin B, liposomal amphotericin B (three dosages), fluconazole, and D0870 (a new experimental oral bis-triazole) was assessed in a CF1 mouse model of hematogenous C. krusei infection. Increased survival time and reduced kidney fungal burden were achieved with treatment with amphotericin B at 2 mg/kg/day and liposomal amphotericin B at 8 and 15 mg/kg/day. D0870 at 25 mg/kg/day increased survival time but had no effect on clearance from organs, while the survival and clearance from organs of mice treated with fluconazole at a dose of 100 mg/kg/day did not differ from those of untreated animals. These findings suggest that deoxycholate and liposome-encapsulated amphotericin B are active against disseminated C. krusei infection in neutropenic mice and confirm the in vitro and in vivo resistance of this species to fluconazole. PMID- 8228371 TI - Inhibition of Candida albicans translocation from the gastrointestinal tract of mice by oral administration of Saccharomyces boulardii. AB - Microbial translocation is defined as the passage of viable microbes from the gastrointestinal (GI) tract to extraintestinal sites, such as the mesenteric lymph node (MLN), spleen, liver, kidneys, and blood. The ability of orally administered viable Saccharomyces boulardii to inhibit Candida albicans translocation from the GI tract was tested in antibiotic-decontaminated, specific pathogen-free (SPF) mice, which were orally challenged with C. albicans to promote intestinal overgrowth and subsequent translocation of this organism. Oral S. boulardii treatment reduced the incidence of MLN cultures positive for C. albicans but did not decrease the numbers of C. albicans per gram of MLN in these immunocompetent mice. Prednisolone immunosuppression increased translocation of C. albicans to the MLN and allowed translocating C. albicans to spread systemically to the spleen, liver, and kidneys. In these immunosuppressed mice, orally administered S. boulardii decreased both the incidence of C. albicans translocation to the MLN, liver, and kidneys and the number of translocating C. albicans per gram of MLN, spleen, and kidneys. PMID- 8228372 TI - Degradation of human IgA by Entamoeba histolytica. AB - To determine whether the virulent enteric pathogen Entamoeba histolytica degrades human IgA molecules, serum and secretory IgA was exposed to viable axenic trophozoites (strain HM1:IMSS), a parasite sonicate, and medium conditioned by incubation with live trophozoites. IgA was completely degraded under all conditions, proteinase activity was maximal at a neutral pH, and there was a four to eightfold enrichment of amebic IgA proteolytic activity in a soluble fraction of amebic sonicate. Degradation of serum IgA by amebic sonicate was completely inhibited by the cysteine proteinase inhibitors trans-epoxysuccinyl-L-leucyl amino(4-guanidino)butane (E-64, 100 microM) and benzyloxycarbonyl-phenyl-alanyl alanyl-fluoromethyl (Z-Phe-Ala-CH2F, 12.5 microM). Secretion of degradative activity, the optimal pH, and the inhibition by E-64 and Z-Phe-Ala-CH2F indicates that cysteine proteinase activity is predominantly responsible for the degradation of human IgA by E. histolytica. PMID- 8228373 TI - Detection of Plasmodium vivax by polymerase chain reaction in a field study. AB - Detection and typing of Plasmodium vivax by the polymerase chain reaction (PCR) was evaluated in a prospective blinded comparative field study in Thailand. PCR amplification of the circumsporozoite (CS) gene was compared with microscopy for the detection of P. vivax in blood samples from 174 Thai Rangers and 50 malaria free Bangkok residents. For PCR analysis, filter paper specimens collected by finger prick were randomly processed and blindly interpreted for the presence of the CS gene of P. vivax. The VK210 and VK247 CS variants of P. vivax were detected by specific fluorescein or radiolabeled oligoprobes. Autoradiography with 32P-labeled probes and enhanced chemoluminescent detection with fluorescein labeled probes identified 91% and 96%, respectively, of 119 microscopically confirmed infections; both systems detected < 100 parasites/microL. Compared with microscopy, the specificity of PCR and radiometric or enhanced chemoluminescent detection was 96% and 90%, respectively. The ease of collection and transport of filter-paper specimens combined with the sensitive and specific detection of allelic genes of P. vivax by PCR suggests that this method may prove to be a valuable tool for epidemiologic and heterogeneity studies of P. vivax. PMID- 8228374 TI - Tumor necrosis factor-alpha in human immunodeficiency virus-infected patients: correlations with opportunistic infections and immunologic status. PMID- 8228375 TI - Hepatitis C virus genotypes in chronic hepatitis and response to interferon-alpha therapy. PMID- 8228376 TI - Cytomegalovirus resistant to foscarnet: clinicovirologic correlation in a patient with human immunodeficiency virus. PMID- 8228377 TI - Meningoradiculoneuritis due to acyclovir-resistant varicella-zoster virus in a patient with AIDS. PMID- 8228378 TI - Single-dose versus 7 days of fluconazole treatment for oral candidiasis in human immunodeficiency virus-infected patients: a prospective, randomized pilot study. PMID- 8228379 TI - Opportunistic infections and profound CD8+ T lymphocytopenia in mixed connective tissue disease. PMID- 8228380 TI - Promotion of transposon Tn916 excision via exposure to streptomycin. PMID- 8228381 TI - Detection of Leptospira DNA by polymerase chain reaction in aqueous humor of a patient with unilateral uveitis. PMID- 8228382 TI - The digestive tract is a major site for Acinetobacter baumannii colonization in intensive care unit patients. PMID- 8228383 TI - Rapid identification of Mycobacterium avium-intracellulare complex strains: clinical practice evaluation of DT6 and DT1 probes. PMID- 8228384 TI - Microbial cell walls and resistance of bacteria and fungi to antibiotics and biocides. PMID- 8228385 TI - Soluble tumor necrosis factor receptors in Plasmodium vivax malaria. PMID- 8228386 TI - Interferon production by and leukocyte phenotyping of rhesus monkey lymph node and peripheral blood cells. AB - The ability of peripheral blood leukocytes to produce interferon-gamma (IFN gamma) and be labeled with monoclonal antibodies against cell-surface markers was determined in this study. Both peripheral blood leukocytes and lymph node cells were able to produce IFN-gamma after challenge with mitogens. The rhesus monkey IFN-gamma was detectable by means of a biological assay but not by means of a radioimmunoassay for human IFN-gamma. Peripheral blood leukocytes and lymph node cells from rhesus monkeys (Macaca mulatta) were treated with fluoresceinated antibodies directed primarily against cell-surface antigens of humans. The degree of binding was determined by means of flow cytometry. Several of the anti-human antibodies did bind to the rhesus monkey peripheral blood leukocytes, as expected. In a novel study, the antibodies bound in a similar fashion to rhesus monkey lymph node cells. Binding of the antibodies was equivalent whether the cells came from inguinal or axillary lymph nodes. Rhesus monkey peripheral blood leukocytes incubated with recombinant human IFN-gamma showed enhanced expression of class II major histocompatibility complex antigens, as detected with anti-HLA DR antibodies. PMID- 8228387 TI - Effect of carbohydrates on the pharmacokinetics of human interferon-gamma. AB - Human interferon-gamma (IFN-gamma) has two N-linked glycosylation sites at positions 25 and 97 of the 143-amino-acid-long secretory form. To study the role of glycan residues in the pharmacokinetics of IFN-gamma, we produced recombinant IFN-gamma molecules lacking either one or both of the glycosylation sites (Asn mutated to Gln) by baculovirus expression in insect cells. In addition, we produced the fully glycosylated forms both in insect cells and in human leukocyte cultures. Two million units of each IFN were injected intravenously or intramuscularly into rabbits. The glycosylated IFN-gamma molecules from the insect cells were rapidly eliminated from the blood. This is probably due to the fact that their oligosaccharides are of a high mannose type that are rapidly taken up by the liver. The unglycosylated IFN-gamma persisted longer in the blood than the glycosylated recombinant forms. However, the natural IFN-gamma exhibited the longest survival in the blood. The results emphasize the importance of the carbohydrate groups in human IFN-gamma to its pharmacokinetic properties. PMID- 8228388 TI - N-acetyl cysteine enhances the response to interferon-alpha in chronic hepatitis C: a pilot study. AB - Hepatitis C virus (HCV) is an RNA virus that replicates in both the liver and lymphoid cells. Interferon-alpha (IFN-alpha) is a useful treatment of chronic hepatitis C (CHC) although resistance to this drug occurs frequently. The mechanisms underlying resistance to IFN remain unknown. In this work, we have measured the levels of glutathione in plasma and peripheral lymphoid cells from 15 healthy controls and 24 CHC patients, 10 of whom were without treatment and 14 showed high serum alanine aminotransferase (ALT) values despite therapy with lymphoblastoid IFN for more than 4 months. In all patients, glutathione levels in plasma and in mononuclear cells were depressed in comparison to controls. In IFN unresponsive patients, the addition of 600 mg tid of oral N-acetyl cysteine (NAC), a glutathione precursor, resulted in a steady decrease of ALT values in all patients, with complete normalization in 41% of cases after 5-6 months of combined therapy. Administration of NAC alone for 1 month was without effect in the 10 patients that were not receiving IFN. Supplementation of IFN with NAC induced a near normalization of intralymphocytic glutathione, but plasma levels were only moderately increased. HCV replication was markedly inhibited in lymphocytes and viremia was cleared in one of the 8 patients tested. In conclusion, NAC enhances the response to IFN in CHC. Controlled studies are needed to ascertain whether antioxidant therapy might act in synergy with IFN in chronic viral hepatitis. PMID- 8228389 TI - Interferon-alpha and interferon-gamma induced modulation of proteins in human corneal fibroblasts. AB - Little is known about the effects of interferon (IFN) on cell function in the eye. We have analyzed the effect of INF-alpha and IFN-gamma on the expression of proteins in cultured human corneal fibroblasts. Treatment with IFN-alpha increased the synthesis of proteins of 84, 76, 52, and 28 kD and decreased the synthesis of a 72-kD protein. Treatment with IFN-gamma increased the synthesis of proteins of 83, 66, 64, 54, and 47 kD. The effect of IFN-alpha and IFN-gamma were first detected at 5-9 h and 9 h, respectively, after the addition of the IFNs and were maximal at 17 and 24 h, respectively. Most of the changes were seen at doses of 1 x 10(1) to 1 x 10(2) U/ml of IFN-alpha or IFN-gamma and were maximal at 1 x 10(2) to 1 x 10(3) U/ml. Thus, each IFN induced distinct proteins based on apparent molecular weight and isoelectric point. These results show that IFN alpha and IFN-gamma affect the synthesis of small groups of distinct proteins in human corneal fibroblasts. PMID- 8228390 TI - Recombinant interferon-tau regulates secretion of two bovine endometrial proteins. AB - Conceptus-derived interferon-tau (IFN-tau) has been implicated in the process of maternal recognition of pregnancy in the bovine. This type I IFN interacts with a uterine receptor complex to elicit secondary maternal responses, one of which is secretion of uterine proteins. We investigated the effects of pregnancy and recombinant (r) bovine (bo) IFN-tau (10(7) antiviral units/mg) on secreted proteins by cultured bovine endometrial explants. Explants were cultured for 24 h with [3H]leucine and dialyzed medium analyzed by one- and two-dimensional SDS polyacrylamide gel electrophoresis (PAGE) and fluorography. In one-dimensional PAGE experiments, endometrium representing early pregnancy (days 16-21) and the estrous cycle (days 16-19) was cultured in the presence of 5 nM rboIFN-tau and showed an increase (two- to five-fold) in secretion of 12- and 28-kD proteins. Further examination of these proteins by using two-dimensional PAGE indicated that the 12-kD protein was basic (pI > or = 7.5), whereas the 28-kD protein was acidic (pI approximately 5.0). Isoelectric focusing in the acidic range revealed that the 28-kD protein was composed of several isoelectric variants (pI 4.5-5.5). Although the functions of these secretory proteins are currently unknown, they serve as useful markers for IFN action and may act as secondary signals to protect the early developing conceptus. PMID- 8228391 TI - Nasopharyngeal angiofibroma: a case study. AB - Juvenile angiofibromas, most commonly found in adolescent males, are vascular lesions that occur in the nasopharynx and extend into other areas of the skull. Presenting signs and symptoms include epistaxis, nasal obstruction and nasal drainage. Computed tomography and magnetic resonance imaging are the primary diagnostic tests. Angiography is used for vascular mapping and preoperative embolization. Total surgical resection is the goal of treatment. Nursing care includes developmental considerations with a focus on comfort, neurological assessment and patient and family education. PMID- 8228392 TI - Using a ventricular reservoir to instill amphotericin B. AB - Intrathecal administration of amphotericin B is the best method of eradicating intracranial fungal infections. The Ommaya reservoir provides an easy and practical method for fungicidal medication administration. Treatment of coccidioidomycosis with amphotericin B may be accomplished via an Ommaya reservoir. Astute nursing care is essential to prevent complications associated with this procedure. PMID- 8228393 TI - Glasgow scale relationships in pediatric and adult patients. AB - The purpose of this study was to examine the relationship of the Glasgow Coma Scale (GCS) and Glasgow Outcome Scale (GOS) scores in pediatric and adult patients having surgical entry into the cranium for diagnostic or therapeutic purposes after brain insult. In this retrospective record review, the predictor variable was the coma-associated GCS score (GCS score 3-8) obtained 24 hours after surgery. The outcome variable was the combined Glasgow Outcome category: recovery/moderate disability; severe disability/vegetative state; and death; ascertained 14 days postoperatively. Using discriminant analysis, the unweighted GCS scores for the total sample significantly discriminated two of the three GOS outcomes (p < .001); the weighted scores discriminated all three outcomes (p < .001). Considering the GCS components, the eye scores significantly discriminated one outcome, the motor scores two outcomes, and the verbal scores two outcomes (p < .001 to .005). The unweighted GCS scores in adults significantly discriminated all three outcomes and in children two outcomes; the weighted GCS scores in both children and adults significantly discriminated all three outcomes. The approximately 25% misclassification rate for outcomes, however, would preclude using the scales in a predictive or prescriptive manner. PMID- 8228394 TI - Paraneoplastic syndromes: a challenge for neuroscience nursing. AB - Neurologic paraneoplastic syndromes are medical and nursing challenges. Although neurologic paraneoplastic syndromes are relatively rare in the total cancer population, they may be more commonly seen in the acute neurologic setting. O'Neill et al estimate that 60% of persons with Eaton-Lambert syndrome and 50% of persons with subacute cerebellar degeneration have an underlying cancer. The slow and subtle development of neurologic paraneoplastic disorders underscores the need for careful evaluation of persons with neurologic deficits. Once the diagnosis is made, nursing care must be based on the problems associated with paraneoplastic manifestations of the disease, as well as effects of the cancer and its treatment. PMID- 8228395 TI - Case study of two methods for enteral phenytoin administration. AB - Phenytoin is a medication administered for the treatment or prophylaxis of seizures in head-injured patients. When enterally administered, serum phenytoin levels fall dramatically and are often subtherapeutic. The exact cause for this phenomenon has not been determined. The purpose in this case study was to evaluate the effect of two methods of administering phenytoin suspension in head injured patients receiving concurrent enteral feedings by measuring subsequent serum phenytoin levels. Purposive sampling was used which included two female participants. For participant 1, data revealed dilution of the suspension and irrigation of the tube produced higher phenytoin levels than administering the suspension undiluted. Both serum phenytoin levels were profoundly subtherapeutic in participant 2. Factors such as simultaneous administration of other enteral medications to the second participant might have interfered with phenytoin absorption. PMID- 8228396 TI - Issues in data management and storage. AB - Safeguarding the integrity of the scientific enterprise involves many elements. In this article, issues related to proper management and storage of data have been explored. Such issues have assumed increased prominence in the biomedical research community in the wake of new PHS regulations. Guidelines were proposed to assist nurse researchers, and a national data archive has been recommended for the future. PMID- 8228397 TI - Oral and facial movements in the aged. AB - Orofacial movements are often outward manifestations of many generalized organic processes. When muscle weakness and hyperactivity are present, tumors and lesions at the base of the skull should be ruled out. Treatments involving myotomies and denervations are drastic, but can be effective even if disfiguring. Botulinum toxin offers short-term (weeks to months) relief, and when used sparingly the muscles involved are only modestly impaired. Since the etiology of many of these movements is unknown, treatments are usually only palliative and remissions rare. PMID- 8228398 TI - The year after: post-concussion syndrome. PMID- 8228399 TI - Intrathecal baclofen for spinal spasticity: implications for nursing practice. AB - Treating severe spasticity with the continuous infusion of intrathecal baclofen through an implantable delivery system is a new and successful alternative to oral medications and ablative surgical procedures. Nurses have a vital role in patient care throughout all phases of therapy. Advances in this technology hold promise for our future in treating other neurologic disorders. PMID- 8228400 TI - [Blalock-Taussig shunts in infants less than 6 months of age with special reference to the long-term administration of prostaglandin E1]. AB - Classical Blalock-Taussig (B-T) shunt operation was performed in 14 neonates (group N) and 22 infants (group P). A mean age of group N patients was thirteen days and that of group P patients was two months. Preoperatively prostaglandin E1 (PGE1) was administered in all patients and was continued for a mean of 7 days (range 2 hours to 21 days) in group N, and a mean of 78 days (range 42 to 158 days) in group P. Body weight (4359 +/- 339 g), size of subclavian artery (3.7 +/ 0.5 mm in diameter) and size of pulmonary artery (6.4 +/- 1.2 mm in diameter) at operation of group P patients were larger than those (3212 +/- 490 g, 3.4 +/- 0.4 mm, 5.1 +/- 1.3 mm) of group N. But there were no differences in operation time (165 +/- 22 minutes in group N vs 174 +/- 25 minutes in group P) and clamp time of pulmonary artery (31 +/- 10 minutes in group N vs 30 +/- 5 minutes in group P) between 2 groups. There was no operative death in both groups. There were one late death in group N and three late death in group P. Five patients in group N and nine patients in group P required additional B-T shunt at the mean follow up interval of two years.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228401 TI - [A clinical study on respiratory management using a respirator with nasal intubation in patients undergoing surgery for esophageal cancer]. AB - We analyzed a postoperative respiratory management using a respirator with nasal intubation in 55 patients who underwent subtotal esophagectomy through thoracic and abdominal approach between April 1984 and December 1989. In 21 cases (38%) the period using a respirator was within 3 days, in 24 cases (44%) during 4-7 days, and in 10 cases (18%) over 8 days. Postoperative pulmonary complications occurred in 20 cases (36%); pneumonia in 7 cases (13%), lung edema in 12 cases (22%), atelectasis in one case (2%). Two patient died after surgery, one from acute myocardial infarction, another from multiple organ failure after anastomotic leakage. The period using a respirator was positively correlated with the operative blood loss, transfusion and post-operative infusion, postoperative pulmonary complications positively correlated with the operative time, the anesthetic time, operative blood loss, transfusion and postoperative infusion as well as with preoperative complications and renal dysfunction too. In 120 cases before 1984, postoperative pneumonia occurred in 42 cases (36) and 19 cases (16%) of them died. These results suggest that our respiratory management using a respirator is effective in perioperative period of esophageal cancer. PMID- 8228402 TI - [Noninvasive assessment of left internal thoracic artery graft patency using transthoracic echocardiography]. AB - To evaluate the utility of transthoracic echocardiography (TTE) for assessing the long-term patency of left internal thoracic artery (LITA) grafts, 36 patients who had coronary artery bypass grafting (CABG) were examined simultaneously for graft flow velocity by TTE and by graft angiogram. The flow velocity in the LITA graft was clearly visualized by TTE in 25 patients from the left supraclavicular echo view. On the basis of the angiogram, the sensitivity and specificity of TTE in the diagnosis of LITA graft patency were 71.4% and 100%, respectively. These 25 patients (mean follow-up time after CABG-29.6 months) were divided into 3 groups according to their preoperative diagnosis and the presence of grafts stenosis by angiogram as follows: APS(-), angina pectoris without graft stenosis in 15 cases; APS(+), angina pectoris with graft stenosis in 2 cases; OMI, old myocardial infarction without graft stenosis in 8 cases. The peak flow velocity D/S ratio and time velocity integral D/S ratio in the APS(-) group (2.48 +/- 0.73, 3.21 +/- 0.89) were significantly (p < 0.01) higher than those in the APS(+) group (0.30 +/- 0.23, 0.35 +/- 0.21) or in the OMI group (0.96 +/- 0.44, 0.87 +/- 0.51). Peak flow velocity D/(S+D) and time velocity integral D/(S+D) ratios in the APS(-) group (0.70 +/- 0.05, 0.75 +/- 0.06) were significantly (p < 0.01) higher than those in the APS(+) group (0.22 +/- 0.14, 0.25 +/- 0.11) or in the OMI group (0.46 +/- 0.17, 0.43 +/- 0.18).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228403 TI - [Surgery for thoracic aortic aneurysm by retrograde total body perfusion and deep hypothermia]. AB - We performed retrograde total body perfusion and deep hypothermia for DeBakey III aortic dissection and aneurysm of the distal arch. We could operate with only posterolateral skin incision using a long venous cannula, which is originally a part of percutaneous cardiopulmonary support (PCPS). The duration of retrograde total body perfusion was 31-101 minutes, the perfusion volume 0.6-1.4 L/min, the central venous pressure 5-16 mmHg. All five cases were successfully operated. As a postoperative cerebral accident, one case showed transient dysfunction. There was no postoperative liver and renal dysfunction. The perfusion volume was maintained in 2.5-3.0 L/min during ventricular fibrillation, which is occurred in cooling and rewarming process. So, this long venous cannula worked well to assist circulation. We think retrograde total body perfusion is a useful assisting device for DeBakey III aortic dissection and aneurysm of the distal arch. PMID- 8228404 TI - [The evaluation of interval period to prepare the left ventricle for Jatene operation for simple transposition of the great arteries]. AB - Laboratory studies indicate that it can be as short as one week after a first stage operation to prepare the left ventricle for a subsequent second-stage arterial switch (Jatene operation) for simple transposition of the great arteries (TGA). In order to ascertain it, we evaluated the left ventricular (LV) function immediately after the first-stage procedure and late postoperatively by echocardiography and catheterization. There were 6 cases with simple TGA. The age at palliation was 6.6 +/- 4.5 (mean +/- SD) months, and the interval between the first-stage and Jatene operation was 16 days-16 months (6.7 +/- 6.4 months). All patients survived after arterial switch. In 2 early infant cases, arterial switch operation was performed with ventilatory support due to cardiac and respiratory failures 16 and 21 days after the first-stage operation respectively. LV end diastolic dimension increased by 1 week and showed no change after 1 week (n = 4). LV ejection fraction was depressed from 0.84 +/- 0.03 preoperatively to 0.61 +/- 0.08 at 1-4 days after the first-stage operation but improved to 0.83 +/- 0.06 by 7-9 days after that (n = 4). LV end-diastolic posterior wall thickness increased to more than 4 mm in 3 infant cases, more than 3 mm in 2 early infant cases by 7 days. The results of catheterization after the first-stage operation were as follows (n = 5); left ventricular right ventricular peak systolic pressure ratio increased from 0.53 +/- 0.16 to 0.94 +/- 0.04 (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228405 TI - [The relation between the time and the fatigue of skeletal muscle ventricle]. AB - It is generally recognized that if skeletal muscle is stimulated corresponding with the heart rate to assist the cardiac performance, the contraction power of the skeletal muscle decreases in a few minutes. It becomes necessary that the skeletal muscle is electrically stimulated in 6-9 weeks to lead into the transformation or that the muscle is laid without working after operation in 6-9 weeks to wait for growing of the collateral vessels. The observation time of the fatigue phenomenon in many studies is, however, short and it is not understandable that the contraction power decreases near to zero although the type I fibers remain. So we studied the relationship between the time and the pressure assist of the skeletal muscle ventricle (SMV). It was concluded that 1) the cause of the power decrease is not only the fatigue but the operative invasion. 2) in our setting, when SMV was stimulated in the ratio of 1:4 or 1:10 to the heart rate, the power recovered within 3 hours. 3) if the initial setting of the drive is light enough, the preconditioning or the vascular delay will be unnecessary and the working transformation or the in situ training will practicable. PMID- 8228406 TI - [Surgical results in atherosclerotic distal aortic arch aneurysm]. AB - Results of surgical treatment of atherosclerotic aneurysm of the distal aortic arch, reported to date, are not satisfactory. We recently analyzed the surgical results in 119 patients with this condition who were treated at our department. There were 104 males and 15 females, ranging in age from 38 to 81 years (mean: 66 years). The arch was reconstructed by total replacement in 29 cases, partial replacement in 49 cases, patch reconstruction in 34 cases and extra-anatomic bypass in 7 cases. The lesion was approached by median sternotomy in 75 cases, median sternotomy+left thoracotomy in 9 cases and left thoracotomy in 35 cases. In cases where median sternotomy was used, selective cerebral perfusion was generally employed as an auxiliary procedure. After surgery, cerebral complications were observed in 19 cases (16.0%). In 15 of these 19 cases, postoperative cerebral damage was attributed to atheromatous emboli. Twenty-six patients (21.8%) died in the hospital. Major causes of death included respiratory insufficiency, multiple organ failure and low cardiac output syndrome. To improve surgical results for this type of aneurysm, the operative procedure needs to be modified so that these postoperative complications are prevented. Especially it is essential to prevent intraoperative embolisms of brain and coronary arteries, and a reconstruction by total arch replacement with median sternotomy has improved recent surgical results. PMID- 8228407 TI - [An experimental study of coronary arteriosclerosis after heart transplantation]. AB - The purpose of this study is to evaluate the effects of Cyclosporine (CsA), FK 506 (FK) and 15-Deoxyspergualin (DOS) on coronary arteriosclerosis after rat heart transplantation. Three groups of Lewis rats (n = 7, each) received heterotopic heart transplants from F-344 donors and were treated with CsA (Group Cs), FK (Group FK) and DOS (Group DOS) intraperitoneally. All rats were sacrificed 60 days later and assessed microscopic grading score (GS) of rejection and arteriosclerosis, and measured serum lipid. There was no significant difference in the GS of rejection but the GS of arteriosclerosis in the groups Cs was significantly higher than the group DOS (1.71 +/- 0.24 versus 1.11 +/- 0.34, p < 0.01). Triglyceride in the groups Cs was significantly higher than the group FK and group DOS (99 +/- 23 versus 66 +/- 21, 56 +/- 30 mg/dl, p < 0.02), and LDL in the group Cs and group FK was significantly higher than group DOS (104 +/- 17, 81 +/- 23 versus 31 +/- 17 mg/dl, p < 0.01). We concluded that DOS had a superior protective effect against coronary arteriosclerosis after heart transplantation and it may depend on the different mechanism of immunosuppression and lipid metabolism abnormality causing by immunosuppressants. PMID- 8228408 TI - [Post operative embolism and left atrial enlargement in the old-aged ASD (atrial septal defect) patients]. AB - As the operation risk of ASD is little today, the unexpected postoperative complication such as embolism is a miserable concern. We experienced two cases of embolism, which were cerebral infarction, in old-aged patients after ASD correction. It has been reported that low cardiac function, atrial fibrillation and foreign body in the left atrium should be risk factors of the embolism after ASD correction. However, blood stasis in the left atrium due to its enlargement may be an essential risk factor of the embolism. Accordingly we evaluated the left atrial volume in 87 ASD patients without complicated cardiac disorder. There was a significant correlation between the age (X) and left atrial volume index (Y). (Y = 16.5 + 1.1X, r = 0.668, p < 0.001). Left atrial volume index in our two cases of cerebral infarction indicated 171.7, 190.8 ml/m2 respectively. The former case had sinus rhythm and its defect was directly closed, and the latter had atrial fibrillation and its defect was closed with teflon patch. In addition pulmonary artery pressure and cardiothoracic ratio correlated with left atrial volume index significantly. Although some investigators reported that pulmonary hypertension and cardiomegaly were the risk factors of embolism, the left atrial enlargement by aging was thought to be the main risk factor. Thus, we concluded that anticoagulant therapy was necessary for the ASD patients with marked left atrial enlargement. PMID- 8228409 TI - [Angiographic evaluation of reconstructed spinal arteries in thoracic aortic aneurysm surgery]. AB - Revascularization of the spinal arteries with thoracic aortic aneurysm were performed on nineteen patients using partial extracorporeal bypass. They were 16 men and 3 women. Age range were from 33 to 70 years (mean 49.9 +/- 10.3 years). There were eleven patients of dissecting aneurysm (DeBakey type IIIb in eight patients, type I in two patients and type IIIa in one patient), and eight patients of non-dissecting thoracoabdominal aneurysm (including two patients with ruptured aneurysm). The number of revascularized spinal arteries were 60 pairs (average 3.2 pairs per each patient). The revascularized spinal arteries were localized between levels T4 and L5.36 pairs of the 60 existed between levels T8 and L2 from where the artery of Adamkiewicz arises. Seven patients (eleven spinal arteries) underwent selective angiography of the revascularized spinal arteries postoperatively, and the anterior spinal artery and the artery of Adamkiewicz was identified in three patients. Two patients died within one month, one from MOF and the another from intestinal perforation respectively (operative mortality 11.1%). One patient, with ruptured thoracoabdominal aortic aneurysm showed paraparesis postoperatively, but no paraplegia was found in any patients. We recommend that not only the artery of Adamkiewicz but also the spinal arteries at the midthoracic area from T4 to T8 should be revascularized, to prevent postoperative paraplegia. Replacing of extended thoracic aneurysm, our method (using partial extracorporeal circulation and segmental aortic clamping) was thought to prevent spinal cord ischemia. PMID- 8228410 TI - [Experimental study of ventricular assist device incorporated with a membrane oxygenator]. AB - We designed a new device that incorporated a VAD out let with a membrane oxygenator, and examined the result. In the mock circulation study, a sufficient inlet flow from the membrane oxygenator for oxygenation and removal of carbon dioxide was obtained. Sufficient flow and pressure to maintain systemic circulation were also obtained. In the hypoventilation and heart failure models' study, with this device, hemodynamics were improved and recovered from heart failure. This device was also able to maintain systemic circulation. And the data of blood gas analysis were also improved. These results suggest that this new assisted-circulation system is possibly a superior circulation assist method which has circulatory assist effects and respiratory assistance capabilities. Moreover, we suspect that these findings can be put to use in clinical cases that include heart failure associated with respiratory failure. PMID- 8228411 TI - [Clinicopathologic studies on esophageal carcinoma with basaloid features]. AB - Eight unusual esophageal carcinomas chiefly composed of basaloid cells were defined as "esophageal carcinomas with basaloid features", and analyzed by clinicopathologic findings and immunohistochemical staining with antibodies against S-100, actin, vimentin, and cytokeratin. These tumors were classified into five basaloid-squamous cell carcinomas (BSC), 2 adenoid cystic carcinomas (ACC), and 1 non-small cell type-undifferentiated carcinoma based on the Guidelines for the Clinical and Pathologic Studies on Carcinoma of the Esophagus edited by the Japanese Society for Esophageal Disease. Some histologic patterns of BSC, ACC, or undifferentiated carcinoma were resembled to each others. However, BSC was characterized by solid growth of basaloid cells with microcystic pattern or peripheral palisading arrangement, and it showed simultaneous presence of squamous cell carcinoma or squamous differentiation. In immunohistochemical staining, immunoreactivity for cytokeratin alone was positive in one of five BSCs. On the other hand, ACC showed cribiriform pattern and high mitotic activities, excluding typical cribriform pattern in ACC of the salivary gland. In addition to these features, the presence of foci of squamous cell carcinoma was occasionally found and positive immunoreactivities for S-100 and actin were recognized. Therefore, the immunohistochemical staining was useful for distinction among ACC and other tumors. As to the prognosis, BSC and ACC have been reported that both tumors are aggressive neoplasm with poor prognosis. In our studies, all of 5 patients with BSC were survived more than three years after surgery. In contrast, the patients with ACC or undifferentiated carcinoma died within one year.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228412 TI - [Cardiac function following left ventricular aneurysm repair--comparison between patch reconstruction and direct closure methods]. AB - Left ventricular function was compared between two groups of patients who underwent patch reconstruction or direct closure of the ventricular wall following resection of a post-infarction left ventricular aneurysm (LVA). There were 15 and 9 patients in the respective groups. Coronary artery bypass grafting was additionally performed in 8 patients (53%) in the patch reconstruction group and 5 patients (56%) in the direct closure group. The size of the patch used for patch reconstruction (including the suture line) was 57 +/- 19% of the resected area. Before and 1-2 months after surgery, equilibrium RI angiography and cardiac catheterization were performed to assess the following hemodynamic parameters: ejection fraction (EF) at rest and during exercise, left ventricular end diastolic pressure (LVEDP), left ventricular end-diastolic volume index (LVEDVI), cardiac index (CI) and mean pulmonary artery pressure (m-PAP). Preoperatively, there were no differences in each parameter between the two groups. Postoperatively, there were significant improvements in the EF at rest and during exercise (p < 0.01), LVEDVI (p < 0.01) and the LVEDP (p < 0.05). However there no significant changes in CI or m-PAP following the operation. There were also no significant differences observed in the postoperative values between the two groups. However, based upon postoperative left ventriculograms, patch reconstruction was significantly superior to direct closure for reconstruction of a septal aneurysm. PMID- 8228413 TI - [Post-sternotomy mediastinitis treated by omental transposition]. AB - Mediastinitis after cardiac surgery is a serious complication with high morbidity and mortality rates. Between December 1989 and April 1992, of 264 patients who underwent cardiac or aortic surgery with median sternotomy, seven (2.6%) developed deep sternal infections with mediastinitis. Six of these patients were treated with debridement, irrigation and delayed wound closure with mediastinal transposition of the greater omentum. Most of the infections were associated with several predisposing factors: cardiac cachexia in the preoperative state, prolonged perfusion time, and respiratory insufficiency in the postoperative period. One of the six patients died of adult respiratory distress syndrome in the postoperative course, but the remaining five patients were discharged with their wounds well healed. Acute and deep mediastinal infection occurred in one patient on the fifth day after cardiac surgery. The infection was successfully treated with radical debridement, two days of intermittent mediastinal irrigation and staged wound closure with omental transposition into the mediastinum. The omental transfer was achieved as follows: after the greater omentum was divided into two pedicles one pedicle was used to fill in the pericardial space and the other was placed between the sternum and the undermined skin. We conclude that early debridement and omental flap transposition are effective in the management of mediastinitis after cardiac surgery. PMID- 8228414 TI - [Effect of retrograde cerebral perfusion in preserving cerebral function during circulatory arrest]. AB - The efficacy of retrograde cerebral perfusion (RCP) method was evaluated in 26 patients who underwent aortic arch reconstruction. Surgical procedures were; ascending-proxymal arch replacement involving Bentall type procedures (17 cases), total arch replacement (4 cases), and distal arch replacement (5 cases). The RCP were performed through selective cannulation on the superior vena cava at 18.4 degrees C of the average rectal temperature. The perfusion pressure was 28.3 (+/- 5.9) cmH2O and flow was 360 (+/- 15.4) ml/min. RCP time was 74.1 (+/- 26.2) min. Twenty-four patients survived among 26 patients and 2 patients died after operation. Postoperative consciousness was confirmed in 25 patients. Postoperative neurological complications were 1) delayed arousal 5 cases, 2) delirium 3 cases, 3) respiratory disorders 5 cases, 4) double vision 2 cases, 5) cerebral infarction (CI) 1 case. These complications except-CI were resolved in early postoperative period. And regarding effective time period of RCP, incidence of neurological complications were significantly higher in cases which received RCP over 80 min. Therefore we consider that 80 min is a safety time period. In conclusion, RCP is a useful step for preserving the brain during the operation of graft replacement of aorta involving aortic arch. PMID- 8228415 TI - [Rastelli procedure for 32-year-old male with truncus arteriosus--a case report]. AB - A 32-year-old male with a diagnosis of truncus arteriosus (Van Praggh Type A3) underwent successfully a total correction by the modified Rastelli procedure. The bilateral pulmonary arteries (PA) were disconnected from the truncus, and the ventricular septal defect was rerouted with a Dacron patch from the left ventricle to the true aorta. A new outflow from the right ventricle to the bilateral PA was established by the use of a T tube graft which consisted of a Rygg's valved conduit and a Rygg's pericardium. The patient had recent cardiac catheterization with slight elevated pulmonary hypertension (Rt-PA 40/10 mmHg, Lt PA 30/10 mmHg), but is doing well at 5 years postoperatively. PMID- 8228416 TI - [Reconstruction of the pulmonary outflow tract without prosthetic conduit for correction of tetralogy of Fallot with pulmonary atresia]. AB - We reconstructed right ventricular outflow tract without prosthetic conduit for 2 year 11 month and 59 day old Tetralogy of Fallot with pulmonary atresia patients. Left appendage anastomosed between pulmonary trunk and right ventricle was utilized as the posterior wall of the tract. Pericardial patch covered the tract the beneficial methods for patients with Tetralogy of Fallot with pulmonary atresia to avoid late postoperative deleterious complications of prosthetic conduit. PMID- 8228417 TI - [Anatomic correction without extracardiac conduit for Taussig-Bing malformation (REV procedure)]. AB - A 2-year-old boy with Taussig-Bing malformation with mild pulmonary stenosis underwent anatomical repair without extracardiac conduit (REV procedure). He had pulmonary arterial banding at 4 months of age, and REV procedure was performed at 2 years and 4 months of age. Under cardiopulmonary bypass, enlargement of ventricular septal defect were performed with an internal conduit. Pulmonary arterial trunk was divided and the proximal end was closed. Direct anastomosis of the posterior wall of the distal pulmonary arterial trunk to the right ventriculotomy was carried out without transection of aorta or translocation of pulmonary artery. Right ventricular outflow tract (RVOT) was reconstructed with a patch bearing a monocusp. The postoperative course was very smooth and postoperative cardiac catheterization revealed no stenosis in both left and right ventricular outlfow tract and good cardiac performance. PMID- 8228418 TI - [A case of constrictive pericarditis following graft replacement of the descending aorta under left ventricular bypass using a centrifugal pump]. AB - We experienced a case of constrictive pericarditis following replacement of the descending aorta. It has been pointed out that constrictive pericarditis can be developed following LV bypass using centrifugal pump due to the cardiotomy. But no actual case has reported so far. The case was a 61-year-old man for whom graft replacement of the descending aorta through left thoracotomy was performed. Fever and heart failure was elicited 3 weeks after the operation, and he was diagnosed as constrictive pericarditis by chest CT and echocardiography. The reoperation was performed 39 days after the graft replacement. Anterior wall of the both ventricle and postero-lateral wall of the left ventricle were strongly adhered to the pericardium. The adhesion was entirely dissected and the thickened pericardium was resected. The post-operative course was uneventful and the patient was discharged 42 days after the reoperation. PMID- 8228419 TI - [A case of papillary fibroelastoma of the heart found 13 years after open mitral commissurotomy]. AB - A patient, 68-year-old female, underwent open mitral commissurotomy (OMC) for rheumatic mitral stenosis in April, 1979. In July of 1992, during the postoperative follow-up echocardiogram showed 8 x 7 mm mitral subvalvular tumor mass and mitral restenosis. Then, mitral valve replacement was performed using prosthetic valve (Carbomedics #25 mm). The tumor was located on the top of postero-medial papillary muscle and its chordae, and histologically was diagnosed as benign papillary fibroelastoma. Based on the findings of periodic echocardiogram, the tumor doubling time was estimated to be 283 days. The postoperative course was uncomplicated and she was discharged on the 34th post operative day. Papillary fibroelastoma is a rare benign cardiac tumor. Pathological origin of this tumor is still unknown. However, Lambl's excrescence, long-term inflammatory stimulation and/or thrombi are supposed to be etiologic factors. This tumor is generally found incidentally at the time of autopsy or operation because the patients are often asymptomatic. However, because the tumor causes embolism at times, periodic examination by echocardiogram after operation preserving valves is recommended, and surgical resection is required when the diagnosis is established. PMID- 8228420 TI - [A report of two cases of coronary bypass surgery after renal transplantation]. AB - Coronary artery bypass surgery was performed in two patients who had had renal transplantation. They had been taking immunosuppressive therapy. There were some problems in the management of these patients including prevention of postoperative infection, renal damage due to cyclosporin, and rejection of the transplanted kidney. Immunosuppressive therapy was continued under the meticulous monitoring of blood cyclosporin levels, blood T-cell subsets, markers of infection such as antiviral titers. The anticipated problems were not encountered in both cases. This is the first report in Japan which describes successful coronary artery bypass surgery for kidney transplant recipients. PMID- 8228421 TI - [A case of traumatic hernia of diaphragm showing prolapse of liver with abnormal lobulation into pleural cavity]. AB - A 48-year-old male was admitted to our hospital in 1988 with injury by a traffic accident, resulting in fracture of the right rib, right clavicle, and pelvic. The patient came to us again with a chief complaint of abdominal pain in May, 1991 when digestive tract gas was observed in the right pleural cavity casting a doubt of diaphragmatic hernia. Chest CT scan and liver scintigraphy disclosed prolapse of the colon into the pleural cavity together with a picture on the diaphragm being supposed as parenchyma of the liver. Operation findings showed prolapses of the colon and the omentum through the opening of diaphragm into the pleural cavity and also exhibited prolapse of hepatic parenchyma, supposedly S4, together with gallbladder into the pleural cavity. The case, which had showed no abnormality in the Chest X-ray film before the injury, was conceivably an extremely rate case of traumatic hernia with the right diaphragm showing; abnormality of hepatic lobulation accompanied by diaphragmatic rupture, and prolapse of the liver with abnormal lobulation. PMID- 8228422 TI - [A case of impending rupture of acute aortic dissection with thrombosed false lumen in early phase]. AB - A 73-year-old patient with acute aortic dissection in delayed shock state following sudden onset of chest-back pain and left sided hemothorax was successfully operated. Contrast enhanced CT demonstrated a crescent-shaped low density area in the descending aorta. Aortography showed ulcer-like projection (ULP) with minor leakage into left thorax, however, a flow in false lumen was not visualized. These findings suggested that the false lumen of type III aortic dissection was thrombosed in its early phase. The aneurysm was transected on left heart bypass through left thoracotomy. The entry of false lumen was found in proximal portion of descending aorta, in which fresh thrombus was filled. A graft was anastomosed and reinforced with felt strips in end-to-end. The postoperative course was uneventful. The ULP by aortography provides a possible sign of rupture in clinical course of aortic dissection. PMID- 8228423 TI - [A case report of malignant mesothelioma]. AB - A case of successful surgical removal of a primary malignant pericardial mesothelioma was reported. The patient was a 79-year-old man with a complaint of dyspnea at rest. Chest X-ray study showed enlargement of upper mediastinum. Echocardiogram, computed tomography and MRI revealed a giant mass occupying the pericardial space anterior to the heart with marked pericardial effusion. The right ventricle was severely compressed with this tumor. Total resection of the tumor was achieved because the tumor was localized without invading surrounding tissue. The tumor was 17 x 12 x 10 cm in size and 900 g in weight. Rare growth fashion as a distinct isolated mass was a key of successful outcome. PMID- 8228424 TI - [A case of benign clear cell tumor of the lung--a review of 17 cases in the Japanese literature]. AB - We experienced a case of benign clear cell tumor of the lung. It is a rare and most unusual pulmonary neoplasm. Only 40 cases have been reported in the foreign literatures and 16 cases in the Japanese literatures. A 30-year-old man, who revealed a coin lesion in the chest roentogenogram at the health screening, underwent resection of the right lower lobe. Microscopically, the tumor cells are similar to clear cell carcinoma of the kidney and have glycogen in the cytoplasm. Ultrastructually, fibrous banded collagen structures were found in the intercellular space. These findings suggest possibility of neural tissue origin of the clear cells. PMID- 8228425 TI - [A five-year survivor of small cell lung carcinoma stage IIIB treated with surgical resection and an adjuvant chemoradiotherapy--a case report]. AB - A 56-year-old woman was admitted to our hospital with the complaint of cough. She had a rt. supraclavicular lymph node swelling, and her chest X-ray showed masses at the rt. hilum and the mediastinum. On bronchofiberscopy, a tumor was detected at the orifice of the lt. B3b, and the biopsy revealed small cell lung cancer (SCLC). With the peroperative diagnosis of stage IIIB SCLC, a neoadjuvant chemotherapy followed by surgical resection was performed. Because of the presence of malignant cells in the pericardial effusion, the operation turned out to be an absolutely non-curative one. As the recurrence of mediastinal lymph node swelling occurred after the surgery, an intensive chemoradiotherapy was performed successfully. Then the patient has been free from disease for five years. This case indicates the possibility that some of SCLC patients can be long-term survivors by the treatment of an appropriate adjuvant therapy combined with radical resection, even if the clinical stage is advanced. Therefore an extended operative indication for SCLC should be considered. PMID- 8228426 TI - [Congenital bicuspid aortic valve with regurgitation--a rare case showing a fibrous band between the conjoined cusp and the ascending aorta]. AB - It is quite unusual that the congenital bicuspid aortic valve shows aortic valve regurgitation except postinflammatory event. In a case of aortic regurgitation with bicuspid valve, a fibrous band was recognized between the conjoined cusp and the ascending aorta at the operation. Since a degenerative change alone was discerned histologically in the resected specimen, this abnormal band was supposed to be an embryogenic remnant of the aortic valve. Six cases were reported previously and all of them were male, and five were Japanese. PMID- 8228427 TI - Diagnosis and treatment planning of hypodivergent skeletal pattern with clockwise occlusal plane rotation. AB - Diagnostic treatment planning of Class III malocclusion is complicated due to the interaction of development, timing, dental pattern, and skeletal relationships. Historically, the treatment of adult Class III malocclusions has tended to be seen primarily as cases of mandibular prognathism. As diagnostic acumen and surgical skills improve and merge, orthodontic and surgical treatment plans for Class III malocclusions have become more common. Class III malocclusions now tend to be described in terms of mandibular prognathism, horizontal maxillary hypoplasia, maxillary vertical hypoplasia, and mandibular dentoalveolar retrusion. The purpose of this paper is to present a combined surgical orthodontic treatment approach involving clockwise rotation of the occlusal plane for treatment of hypodivergent Class III skeletal pattern and dental malocclusion. PMID- 8228428 TI - Changes in tongue and hyoid bone positions following anterior mandibular subapical osteotomy in patients with Class III malocclusion. AB - In a sample of 28 Chinese adults (aged 24.2 +/- 3.1 years) who underwent anterior mandibular subapical osteotomy for the correction of Class III malocclusions, the interaction among intermaxillary space, tongue position, and hyoid bone variables were quantified retrospectively using pretreatment and posttreatment lateral cephalograms. Lateral cephalograms taken at the early (5 to 7 days) postsurgical period showed that the intermaxillary space decreased. The hyoid bone also moved inferiorly, while the tongue was displaced posteriorly and superiorly. The late (2 years) postsurgical lateral cephalograms showed that the hyoid bone and tongue position normalized to the pretreatment positions. These changes demonstrate the ability of the hyoid bone and tongue structures to adapt to changes in the intermaxillary space following osteotomy. PMID- 8228429 TI - Comparison between uniaxial pull-out tests and torque measurement of 2.0-mm self tapping screws. AB - This study investigates the retentive capacity of 2.0-mm self-tapping screws in porcine rib utilizing axial pull-out tests as well as torque measurements of insertion and stripping. These techniques have been performed independently; this analysis assesses the association between pull-out and torque values. The Goodman Kruskel correlation coefficient between pull-out and insertional torque is 0.13, while the correlation between pull-out and stripping torque is 0.21. Partial correlation indexes control the thickness of bone, a confounding factor. Nonparametric analysis of covariance showed no significant difference between insertion of screws and pull-out of the screws, but stripping versus pull-out test was significant. Given these findings, it appears that insertional torque may be used as an alternative technique for screw retention analysis. Because insertional torque tests can be conducted in the operating room, results of clinical studies can be compared to laboratory analyses. PMID- 8228430 TI - A conservative orthodontic-prosthetic approach for a difficult clinical situation: a case report. AB - The accidental loss of maxillary teeth, without replacement, led to an overeruption of the opposing mandibular teeth. The resulting reduction of interarch space created a severe prosthetic problem that was resolved by orthodontic management. PMID- 8228431 TI - Rigid fixation: where do we go from here? PMID- 8228432 TI - Quantitative evaluation of the soft tissue profile in the planning of orthognathic surgery. AB - The clinical application of a proportional soft tissue profile analysis based on natural head position is presented. The analysis provides data concerning the morphological basis of facial disharmony in individual cases. The use of this information during treatment planning and evaluation of treatment results after orthognathic surgery is demonstrated in four patients with severe deviation from midnormal profiles. PMID- 8228433 TI - Skeletofacial changes by mandibular advancement using sagittal split osteotomies. AB - Forty-five patients with mandibular retrognathism treated by surgical mandibular advancement by sagittal split osteotomies were evaluated cephalometrically before treatment at the intermaxillary fixation release, and 6 months postoperatively. The treatment resulted in appreciable improvement of the mandibular retrognathism, and the facial morphology was changed positively toward normal values. The mandibular prognathism was accomplished by an anterior displacement of the mandibular corpus and by a slight increase of the mandibular corpus length. The anterior facial height and the mandibular plane angle were increased by a clockwise rotation of the anterior segment and a counterclockwise rotation of the posterior segment, which consequently increased the gonial angle. These changes partly reversed during the postfixation period. Dentoalveolar changes were found in the position of the maxillary and mandibular incisors; both were uprighted, the mandibular incisors more than the maxillary incisors. In general, the long-term positive effects of the surgical mandibular advancement revealed great stability. PMID- 8228434 TI - A validity test of cephalometric variables as a measure of clinical applicability in anteroposterior profile assessment. AB - Twenty-six profile silhouettes are divided by five judges into three categories: retrognathic, orthognathic, and prognathic. On the corresponding lateral cephalograms, 51 cephalometric variables are tested for profile recognition. Discriminant analysis indicates that measurements based on natural head position perform better than comparable variables based on an intracranial reference line. The soft tissue variables (SnPo'-SnTV, Po'-SnTV, ILS-SnTV) are most successful, but no single variable is totally accurate. Skeletal variables relating the maxilla to the mandible (APo, AB, NA-PoA) are more successful than those variables relating one arch to the remainder of the skull (A-FHP, B-NTV, SNA, SNB). A combination of soft tissue and dental variables (SnPo'-SnTV, SnPo'-TMe', 1u-Npo, 1l-NPo) is found to be completely successful. A methodological approach, as applied in this study, can pinpoint the cephalometric variables that are clinically more effective in describing certain facial features. PMID- 8228435 TI - Relationship between host histones and armadillo-derived Mycobacterium leprae. AB - A major protein previously recognized as being primarily associated with the cell walls of Mycobacterium leprae, major wall protein (MWP), is now identified as histoprotein H2b based on N-terminal amino-acid sequencing, electrophoretic comparisons, and several other properties. An avid association between several host/armadillo-derived histones and M. leprae was demonstrated. Since such armadillo-derived M. leprae are the basis of several ongoing vaccine trials, a simple procedure that permits the prompt solubilization and quantification of histones in M. leprae preparations is described. The quantity of histones associated with M. leprae is significant, ranging from 0.6 to 4.8 micrograms of histoprotein H2b per mg of bacteria. PMID- 8228436 TI - Detection of Mycobacterium leprae by the polymerase chain reaction in nasal swabs of leprosy patients and their contacts. AB - Nose swabs from 4 paucibacillary (PB) and 8 multibacillary (MB) leprosy patients and their contacts were tested for the presence of Mycobacterium leprae by two polymerase chain reactions (PCR); 30% of the samples contained inhibitors for the PCR, 1 of 52 (1.9%) swabs and 13 of 164 (7.9%) swabs were positive for M. leprae among contacts of PB and MB patients, respectively. Since this difference is not significant, and some positives were found among contacts of MB patients treated and cured of their infection, it is concluded that the observed infections are community acquired. PMID- 8228437 TI - Armadillos (Dasypus novemcinctus) as a model to test antileprosy vaccines; a preliminary report. AB - The efficacy of two candidate leprosy vaccines, BCG and a mixture of BCG and killed Mycobacterium leprae, was tested in 62 armadillos caught in the wild. The abilities of the vaccines to convert lepromin-negative armadillos to a positive reaction were compared with a group of control animals. Both vaccines upgraded subsequent lepromin skin-test histopathology. The conversion results parallel the protection values obtained in some BCG vaccine trials against leprosy in humans. Before conducting expensive human trials with new antileprosy vaccines, it would be worthwhile first to evaluate them in the armadillo model. PMID- 8228438 TI - Reconstitution of Mycobacterium leprae immunity in severe combined immunodeficient mice using a T-cell line. AB - To test whether Mycobacterium leprae-immune T cells can confer protection against infection with leprosy bacilli, severe combined immunodeficient (SCID) mice were reconstituted with a BALB/c-derived, M. leprae-responsive, T-cell line. Flow cytometric analysis of spleen and peripheral blood cells confirmed reconstitution with T cells. In vitro lymphokine production and the proliferation of spleen cells from the reconstituted animals established that the donor cells had maintained their functional activity for the duration of the study (275 days). The transfer of immune T cells 24 hr before foot pad infection with leprosy bacilli resulted in a profound reduction in M. leprae multiplication, as compared to the nonreconstituted SCID mice. The yield of acid-fast bacilli in the foot pads of SCID mice reconstituted with the M. leprae-immune T cells also was significantly lower than that found in naive BALB/c mice, and at levels previously found only in BALB/c mice that had been immunized effectively. These experiments demonstrate that M. leprae-immune T cells home effectively and control M. leprae infection in SCID mice. PMID- 8228439 TI - Activity of phenazine analogs against Mycobacterium leprae infections in mice. AB - Twenty-five compounds structurally related to clofazimine were tested for their ability to inhibit the growth of Mycobacterium leprae using the kinetic method of drug evaluation in the mouse foot pad model of leprosy. Seven of the phenazine derivatives displayed anti-M. leprae activity comparable to that of clofazimine when administered at a concentration of 0.01% (w/w) in the diet. Three of the compounds, B746, B4087, and B4101, were active when administered at 0.001% in the diet. At a dietary concentration of 0.0001%, B4087 and B4101 were slightly more active than clofazimine, while B746 was less active. In the kinetic method of drug evaluation, greater anti-M. leprae activity of phenazine derivatives was generally associated with greater pigmentation of abdominal fat. Of the compounds which did not cause pigmentation when fed at a concentration of 0.01% in the diet B4090 was the most active. This compound also inhibits the growth of a clofazimine-resistant M. smegmatis strain. PMID- 8228440 TI - Preservation of Mycobacterium leprae in vitro for four years by lyophilization. AB - Although the viability of Mycobacterium leprae suspended in distilled water with or without 10% fetal calf serum was reduced approximately 10(-2) to 10(-4) from that of the starting material during the process of lyophilization, bacilli capable of multiplication in nude mouse foot pads were found in the lyophilized samples stored for 4 years at 4 degrees C. The multiplication rate of the lyophilized bacilli which were suspended in 10% serum-water was much higher than that of the bacilli suspended in water only. On the other hand, no reduction of the viability of M. leprae suspended in 10% skim milk-water was demonstrated during the process of lyophilization as well as storage for 2 years at 4 degrees C. From the results obtained here, it could be suggested that M. leprae might be preserved in vitro by means of lyophilized M. leprae was extremely stable during cryopreservation when the bacilli were suspended in 10% skim milk-water. Therefore, the composition of the solution for suspending the bacilli is definitely critical for the maintenance of M. leprae viability by means of lyophilization. PMID- 8228441 TI - Immunoreactivity of mycobacterial strain ICRC and Mycobacterium leprae antigens with polyclonal and monoclonal antibodies. AB - ICRC, a cultivable mycobacterium, is undergoing clinical trials as an antileprosy vaccine in India. In the present study, we have investigated the crossreactivity between antigens of the mycobacterial strains of ICRC and Mycobacterium leprae using polyclonal and monoclonal antibodies in a radioimmunoprecipitation assay. It was observed that polyclonal anti-ICRC and anti-M. leprae antibodies showed predominant reactivity to a 21-kDa protein of the mycobacterial strain ICRC and the 21- and 14-kDa proteins of M. leprae. Crossreactivity between the antigens of the mycobacterial strains ICRC and M. leprae was established further by using M. leprae-specific monoclonal antibody WML06 (reacting with the 14-kDa protein of M. leprae), which identified the 21- and 14-kDa proteins of the mycobacterial strain ICRC. Thus, our studies demonstrate that the 14-kDa protein of M. leprae, which is known to harbor T- and B-cell epitopes, shares crossreactive antigenic determinants with the 21- and 14-kDa proteins of the mycobacterial strain ICRC. We believe that such proteins may provide important reagents for designing subunit vaccines and for determining skin-test reagents. PMID- 8228442 TI - Ambiguities in leprosy histopathology. AB - This paper presents the percentage of definite or suggestive evidence present in 482 biopsies from different types of leprosy. The presence of acid-fast bacilli (AFB) and nerve involvement were taken as definite features for a diagnosis of leprosy, and infiltration of the dermal appendages, neurovascular bundles and dermis by granuloma cells and lymphocytes were regarded as suggestive signs of leprosy. Using these criteria, all cases were categorized into three groups having definite, suggestive, or no signs of leprosy. The results showed definite and suggestive features in 72.2% and 14.1% of the cases, respectively. The remaining 13.7% had none of these signs. These cases were mostly healed lesions. Large, epithelioid cell granulomas without any nerve element present and healed cases proved difficult for a definite diagnosis. Emphasis is placed on searching for residual nerve elements in AFB-negative sections because this increases the certainty level of the diagnosis. Also, it is suggested that for uniformity of understanding and reporting, terminologies need to be narrowed down and restricted to only definite, suggestive, or no diagnosis of leprosy. PMID- 8228443 TI - Clinical and histopathological correlation in the classification of leprosy. AB - This study reports our observations on the correlation between clinical and histopathological diagnoses of the classification of leprosy. The histopathological classification of leprosy in 1351 cases was done per Ridley Jopling criteria and was compared with the clinical diagnoses of the same cases. These 1351 cases included 79 cases diagnosed clinically as having a "reaction." However, the histopathologists could not detect any evidence of reaction in 16 of these 79 cases (20%). Of the remaining 1272 cases, 68 (5%) were reported as "no evidence of leprosy" by the histopathologists; 37 of these 68 were found to be from the clinically indeterminate type of leprosy. Histopathological and clinical diagnoses of the classification of leprosy coincided in 69% of the cases. Concordance between the clinical and histopathological diagnoses for different types of leprosy was: indeterminate (I) = 36%, tuberculoid (TT) = 50%, borderline tuberculoid (BT) = 77%, borderline (BB) = 26%, borderline lepromatous (BL) = 43%, and lepromatous (LL) = 91%. When some of the types were combined (BT with TT, BL with LL), the overall concordance figure was 76%; concordance for the TT/BT group was 80%, for the BL/LL group it was 93%. Since both TT and BT are considered paucibacillary and LL or BL are considered multibacillary for treatment purposes, differentiating TT from BT or BL from LL is, perhaps, therapeutically irrelevant. However, for classification purposes it appears that the weight given to different signs and/or histopathological parameters for classifying leprosy cases (especially TT, BB and I) needs to be reassessed. PMID- 8228444 TI - Cell-mediated immunity in leprosy; an update. PMID- 8228445 TI - Cutaneous sarcoidosis masquerading as relapsed borderline tuberculoid leprosy? AB - A patient with cutaneous sarcoidosis is presented. The patient was diagnosed initially as borderline tuberculoid leprosy downgrading to borderline lepromatous leprosy and received a full 2 years of World Health Organization multibacillary drug treatment (MDT). Bilateral hilar lymphadenopathy on chest X-ray and cutaneous anergy manifested by negative Mantoux, candidin and trichophytin skin tests suggested the diagnosis of sarcoidosis. Histopathology confirmed this diagnosis. The subcutaneous nodule seen in this patient is a rare type of cutaneous sarcoidosis. PMID- 8228446 TI - Assessment of BCG protective efficacy by case-control studies. PMID- 8228447 TI - Interleukin-1 beta production by peripheral blood mononuclear cells from leprosy patients. PMID- 8228448 TI - Higher specificity in the serodiagnosis of leprosy by combined titration of antiphenolic glycolipid-I and antiphospholipid antibodies. PMID- 8228449 TI - Autoaggressive lepromatous leprosy. PMID- 8228450 TI - Electrocardiographic alterations in lepromatous leprosy patients with concomitant Trypanosoma cruzi infection. PMID- 8228451 TI - Histoid leprosy in early macular lepromatous leprosy; incidental finding or sign of augmented local immunity? PMID- 8228452 TI - Myokymia in leprosy patients. PMID- 8228453 TI - Reduced fingertip blood flow and peripheral dysautonomia in leprosy patients in Iranian Azerbaijan and in Maharashtra, India. PMID- 8228454 TI - [Allergic diseases: progress in diagnosis and therapy. I. New understanding of etiology and physiopathology. 1. Changes in the life environment and allergy. 1) Allergens (effects of urbanization)]. PMID- 8228455 TI - [Allergic diseases: progress in diagnosis and therapy. I. New understanding of etiology and physiopathology. 1. Changes in the life environment and allergy. 2) Environmental factors]. PMID- 8228456 TI - [Allergic diseases: progress in diagnosis and therapy. I. New understanding of etiology and physiopathology. 2. Immune responses and allergic inflammation. 1) Allergic responses and their control]. PMID- 8228457 TI - [Allergic diseases: progress in diagnosis and therapy. I. New understanding of etiology and physiopathology. 2. Immune responses and allergic inflammation. 2) Mast cells and chemical mediators]. PMID- 8228458 TI - [Allergic diseases: progress in diagnosis and therapy. I. New understanding of etiology and physiopathology. 2. Immune responses and allergic inflammation. 3) T cell activation and cytokines]. PMID- 8228459 TI - [Allergic diseases: progress in diagnosis and therapy. I. New understanding of etiology and physiopathology. 2. Immune responses and allergic inflammation. 4) Eosinophils and bronchitis]. PMID- 8228460 TI - [Allergic diseases:progress in diagnosis and therapy. II. Approach to diagnosis and treatment. 1. Selection and application of allergy tests (IgE levels, allergy specific IgE antibodies, skin reactions, provocation tests, and others)]. PMID- 8228461 TI - [Allergic diseases: progress in diagnosis and therapy. II. Approach to diagnosis and treatment. 2. New mode of application of anti-allergy preparations. 1) Anti allergy preparations (chemical mediator release suppressing agents and antagonists)]. PMID- 8228462 TI - [Allergic diseases: progress in diagnosis and therapy. II. Approach to diagnosis and treatment. 2. New mode of application of anti-allergy preparations. 2) Adrenal cortex steroid preparations]. PMID- 8228463 TI - [Allergic diseases: progress in diagnosis and therapy. III. Primary care in allergic diseases: therapeutic guidelines. 1. Bronchial asthma]. PMID- 8228464 TI - [Allergic diseases: progress in diagnosis and treatment. III. Primary care in allergic diseases: therapeutic guidelines. 2. Allergic rhinitis--therapeutic guideline]. PMID- 8228465 TI - [Allergic diseases: progress in diagnosis and treatment. III. Primary care in allergic diseases: therapeutic guidelines. 3. Atopic dermatitis]. PMID- 8228466 TI - [Allergic diseases: progress in diagnosis and treatment. III. Primary care of allergic diseases: therapeutic guidelines. 4. Food allergy]. PMID- 8228467 TI - [Allergic diseases: progress in diagnosis and treatment. III. Primary care of allergic diseases: Therapeutic guidelines. 5. Emergency treatment of allergic diseases]. PMID- 8228468 TI - [Therapeutic guidelines for allergic diseases: discussion]. PMID- 8228469 TI - [Marked efficacy of double filtration plasmapheresis in 3 cases of adult-onset Still's disease]. PMID- 8228470 TI - [Case of Carney's syndrome with primary pigment nodular adrenocortical disease (PPNAD) and pigmented spots of the lips]. PMID- 8228471 TI - [Case of post-transfusion hepatitis B complicated with Guillain-Barre syndrome]. PMID- 8228472 TI - [Case of paragonimiasis developing following consumption of kimuchi- pickled crabs]. PMID- 8228473 TI - [Medical application of transgenic mice]. PMID- 8228474 TI - [Apoptosis and autoimmune diseases]. PMID- 8228475 TI - [Diagnostic tests of blood platelets in patients with platelet disorders]. PMID- 8228476 TI - [Blood platelet disorders]. PMID- 8228477 TI - [Hemorrhagic diathesis due to blood platelet disorders]. PMID- 8228478 TI - [Diagnostic procedures in patients with platelet disorders]. PMID- 8228479 TI - [Thrombocytopenia due to therapeutic drugs]. PMID- 8228480 TI - [Blood platelets and arteriosclerosis]. PMID- 8228481 TI - [Diet, drugs and blood platelet function]. PMID- 8228482 TI - [Congenital blood platelet disorders]. PMID- 8228483 TI - [Application of platelet transfusion and their problems]. PMID- 8228484 TI - [Diagnosis and therapy of megakaryoblastic leukemia]. PMID- 8228485 TI - [Recent trends in therapy in patients with idiopathic thrombocytopenic purpura]. PMID- 8228486 TI - [Clinical study of blood platelet disorders. Discussion]. PMID- 8228487 TI - [A case of severe malnutrition fatty liver due to portacaval shunt]. PMID- 8228488 TI - [A case of symmetric cerebral atrophy seen in both side of temporal lobe following spontaneous remission of encephalitis due to herpes simplex]. PMID- 8228489 TI - [A case of sterno-costo-clavicular hyperostosis associated with Schoenlein-henoch purpura found at the time of surgical treatment of tooth-root cyst]. PMID- 8228490 TI - [A case of Behcet disease with coronary aneurysm, aortic valve insufficiency, and abdominal aortic aneurysm]. PMID- 8228491 TI - [A case of systemic cytomegalovirus infections associated with pathologic bone fracture]. PMID- 8228492 TI - [Recent progress of therapy in patients with Parkinson's disease]. PMID- 8228493 TI - [Morphology and function of blood platelets]. PMID- 8228494 TI - [Production and breakdown of blood platelets in various platelet disorders]. PMID- 8228495 TI - [Recent changes in cross infections]. PMID- 8228496 TI - [Control in use of antibiotics for prevention of cross infections]. PMID- 8228497 TI - [Cross infections in neonatal intensive care units]. PMID- 8228498 TI - [Cross infections in intensive care units and the countermeasure]. PMID- 8228499 TI - [Cross infections in geriatric hospitals and nursing homes and the countermeasures]. PMID- 8228500 TI - [Cross infections due to MRSA and the countermeasures]. PMID- 8228501 TI - [Cross infections due to Staphylococcus epidermidis and the countermeasures]. PMID- 8228502 TI - [Cross infections due to Mycobacterium tuberculosis and the countermeasure]. PMID- 8228503 TI - [Cross infections due to fungi and the countermeasure]. PMID- 8228504 TI - [Hepatitis as cross infections and the countermeasures]. PMID- 8228505 TI - [Countermeasure for AIDS cross infection]. PMID- 8228506 TI - [Management of patients with tracheotomy and prevention of cross infections]. PMID- 8228507 TI - [Management of patients with bedsore and prevention of cross infections]. PMID- 8228508 TI - [Management of patients with urinary catheter and prevention of cross infections]. PMID- 8228509 TI - [Prevention of cross infections using disinfectants]. PMID- 8228510 TI - [Cross infections. Discussion]. PMID- 8228511 TI - [A case of ACTH deficiency with periodic fever]. PMID- 8228512 TI - [A case of farmer's lung seen in north Kyushu for the first time]. PMID- 8228513 TI - [Preoperative diagnosis of mesenteric vein thrombosis; a case report]. PMID- 8228514 TI - [A case of autoimmune hemolytic anemia following ovarian dermoid cyst]. PMID- 8228515 TI - [Two cases of aplastic anemia with hand tremor during treatment with cyclosporin A]. PMID- 8228516 TI - [New coronary angioplasties]. PMID- 8228517 TI - [Acute coronary syndromes--coronary angioscopy]. PMID- 8228518 TI - [Neural mechanisms involved in generation of masticatory rhythm]. PMID- 8228519 TI - [Crossed inhibition of lateral pterygoid motoneurons in guinea pig]. AB - The effects of stimulation of the lateral pterygoid (LP) nerve on the contralateral LP motoneurons (Mns) were investigated in the decerebrate guinea pig. 1. Stimulation of the LP nerve induced a hyperpolarizing potential (HP) in the contralateral LP Mn. The onset and peak latency of this HP were 2.14 ms and 4.10 ms in 11 LP Mns on the average, respectively. The HP was reversed to a depolarizing potential after intracellular Cl- injection, indicating that it mainly consisted of IPSPs. 2. The antidromically evoked field potential (AFP) in the LP Mn pool was depressed by the stimulation of the contralateral LP nerve. The threshold intensity was about 1.2 times the LP nerve threshold. 3. The crossed depression of AFP was significantly diminished after the destruction of the caudal part of the contralateral trigeminal mesencephalic nucleus, but not by the lesion of the trigeminal spinal tract. 4. The most rostral level in the trigeminal spinal tract nucleus, where the HRP-labelled axon terminals were found after its injection into the LP muscle, was located caudally to the lesion of the spinal tract. It is concluded that the muscle spindle afferents from the LP muscle are involved in the crossed inhibition of the LP Mns evoked by the stimulation of the LP nerve. PMID- 8228520 TI - [The expression of osteoblastic features in a SV40 large T antigen immortalized mouse calvarial cell line]. AB - A new cell line (VMCs) was established from the mouse calvarial cells transfected with the plasmid (pMT1-neo) containing a SV40 large T antigen gene and neomycin resistance gene. The VMCs cell has been subcultured weekly for over 100 passages at the ratio of 1:20-1:60. Their population doubling time was about 15 hours. The VMCs cell seemed to be fibroblast-like monomorphic cells by a phase-contrast microscope. The cells were examined and characterized as osteoblastic features by alkaline phosphatase activity, bone formation in diffusion chamber and the expression of mRNAs of type I collagen and osteopontin. The features were as follows: 1. Alkaline phosphatase activity of the VMCs cell was low in growth phase but was increased by the treatment with retinoic acid. 2. Differentiation of the VMCs cell was examined by diffusion chamber implanted intraperitoneally into Balb/c mouse. Implants were sectioned and stained with hematoxylin-eosin, alizarin red S, von Kossa, alcian blue or Azan. In the chambers formation of mineralized matrices and collagenous fibrils were observed. 3. Northern analysis demonstrated the expression of type I collagen and osteopontin mRNAs. Osteopontin mRNA was increased by the treatment with 1 alpha,25(OH)2D3 or retinoic acid. These results suggest that the VMCs cell have some properties of the osteoblast and consider to be useful for the study of bone formation and the biocompatibility test for the dental or medical materials. PMID- 8228521 TI - [Corrosive changes of dental alloys in oral environment--scanning electron microscopic observation and electron probe microanalysis on crown surfaces]. AB - In order to study the changes of the intraoral metals, the surface condition and composition of the crowns removed from mouths were analyzed by SEM and EPMA. Thirty-four samples were categorized into four types of alloys. Corrosive changes were evaluated by comparing the analysis results between the crown surface and its fresh sectioned surface, and also with the data from former studies. 1. Changes observed in the Au alloys were slight compared to the other three types, both in surface feature and composition, except for a slight decrease in Cu. 2. In some Au-Ag-Pd alloys, nearly no dissolution was evident. Composition changes were small, with a slight decrease in Ag and relative increase in Pd. 3. The greatest changes were observed in the Ag-Sn-Zn alloys and Ag-In alloys, even with short intraoral usage period. Generally, their surfaces were rough, with thick layers of organic materials. The dendritic structure was distinct, and dissolution between each dendrite was observed. 4. Many of the Ni-Cr alloys presented dendritic structure with dissolution. Decrease in Ni and relative increase in Si and Cr were observed. 5. Corrosive changes showed a close resemblance to the results obtained by the conventional basic tests. However, there were some features which did not coincide, depending on the types of alloys. PMID- 8228522 TI - [Observations of bone and incisor dentin of rabbits injected with rhodamine labeled albumin by confocal laser scanning microscope]. AB - The confocal laser scanning microscope (CLSM) is the most recently developed microscope using the confocal system and laser light source. This microscope is possible to create optical sectioning images from any depth of the specimen. In this study, the incorporation of rhodamine-labeled albumin (R-ALB) into the bone and incisor dentin was observed by CLSM. Male Japanese white rabbits were given an intravenous injection of R-ALB every two days for 10 days and sacrificed 3 days after the last injection. Transverse ground section of the tibia about 80 microns thickness showed the incorporation of orange-red fluorescence labeling into the endosteal surface as well as the Haversian system of bone. Each labeling line of R-ALB was clearly visible by CLSM compared with the normal fluorescence image. Three-dimensional reconstruction of the sectioning images showed the process of the incorporation of R-ALB into the bone and the complexity of the labeled pattern. R-ALB was incorporated into the incisor dentin and showed the periodic labeling pattern. The high resolution image of the periodic labeling pattern was observed by CLSM. These results indicated that CLSM is a valuable apparatus for studying the fluorescence labeling images of hard tissues. PMID- 8228523 TI - [Cytotoxicity of anaerobic bacteria isolated from infected root canal]. AB - Bacteria in the infected root canal play an important role in the progression of the periapical lesion. The purpose of the present study was to investigate the cytotoxic effects of bacteria isolated from an infected root canal on the periapical lesion. Four obligately anaerobic bacteria were isolated and their sonic extracts (SE) were prepared. The cytotoxic assay of each SE for MC3T3-E1 cell was used with respect to the morphological cell changes and [3H]-thymidine uptake. The following results were obtained: 1. Four obligately anaerobic bacteria were identified as follows: Porphyromonas asaccharolytica (P. a.), Fusobacterium nucleatum (F. n.), Eubacterium lentum (E. l.), and Peptostreptococcus micros (P. m.). 2. The protein concentrations which induced the morphological cell changes differed in each SE. The SE from P. m. caused cell abnormalities at 10 micrograms protein/ml, F. n. and E. l., at 30 micrograms protein/ml, and P. a., at 100 micrograms protein/ml. 3. The inhibition of the incorporation of [3H]-thymidine into DNA was concentration-dependent. The SE of P. a. showed less cytotoxicity than the others at lower concentrations; 3, 10, 30 micrograms protein/ml. The SE from F. n. had more cytotoxicity at higher concentrations; 100, 300 micrograms protein/ml. These results suggest that these bacterial sonic extracts have a potential to cause morphological cell changes and inhibit the cell proliferation in different concentrations. PMID- 8228524 TI - [Drugs used at our department to prevent infective endocarditis]. PMID- 8228525 TI - [Ecology and growth of oral spirochetes]. PMID- 8228526 TI - [Study on an autocrine motility factor-like substance produced by epidermoid carcinoma of the human oral cavity]. PMID- 8228527 TI - [Effect of thermal stress on metal adhesion and adhesive agents to reduce the stress]. PMID- 8228528 TI - [Structure of the origin of the mylohyoid muscle]. PMID- 8228529 TI - Allicin-induced hypotension in rabbit eyes. AB - The intent of this work was to examine the actions of allicin on 1) intraocular pressure (IOP) in normal and unilaterally sympathectomized (SX) rabbits; 2) cAMP accumulation in the rabbit iris-ciliary body (ICB) and cultured nonpigmented epithelial (NPE) ciliary body cells; and 3) 3H-norepinephrine (NE) release by calculating fractional tritium overflow in response to electrical field stimulation (EFS, 5 Hz, 12 V/cm) in isolated, perfused rabbit ICBs. Allicin, one of the active compounds produced by garlic, was evaluated on IOP and it was determined that allicin (1, 2.5, or 10 micrograms), topically, but not the precursor, alliin (10 micrograms), lowered the IOP unilaterally in normal rabbits. Allicin (10 micrograms) reduced the IOP by 6 +/- 1 mmHg (n = 4) in normal rabbits at 2 hrs (maximum response) whereas no change occurred in sympathectomized rabbit eyes. Moreover, allicin (0.01, 0.1, or 1 microM) caused 40, 40, or 52% inhibition, respectively, of 3H-NE overflow in response to EFS. Isoproterenol (ISO, 1 microM) stimulated cAMP accumulation by 3.6 and 9 fold in isolated rabbit ICB and cultured NPE cells, respectively. Allicin (1 microM) had no effect on basal cAMP level while it inhibited ISO-stimulated cAMP accumulation by 40% and 23% in ICB and NPE cells, respectively. This study suggests that allicin lowered IOP, in part, by dual actions at the neuroeffector junction. PMID- 8228530 TI - Lacrimal secretion stimulants: sigma receptors and drug implications. AB - 3H-DTG (1.3-di(2-[5-3H]tolyl)guanidine) or 3H-haloperidol was added to sigma receptors (25 nM) in the presence of 25 nM spiperone and incubated with increasing concentrations of bromhexine derivatives (phenylalkylamines; 10(-9) to 10(-2)M) in membrane homogenate suspensions. IC50 values for two derivatives ranged from 3.2 to 8.8 nM for both radioligands. A preferred derivative, 7A (N,N' dimethyl-2-phenyl-ethylamine), yielded an IC50 of 7.8 nM for 3H-haloperidol but showed much less affinity in displacing 3H-DTG (IC50 = 900 nM). Applying the technic of Bromberg [Exp. Eye Res., 40:313-320, 1985], in vitro protein secretion rates were measured following stimulation of either lacrimal gland slices or isolated, intact lacrimocytes with the compounds. In vitro protein secretion rates exhibit a dose-response relationship with increases in protein release up to a concentration of 10(-8) to 10(-4) M for various derivatives of bromhexine and 10(-4) M for carbachol. By means of Schirmer strips, tear fluid was collected over a five minute period at 10 and 60 minutes post-dosing following the topical application (50 microliters) to the right eye of New Zealand white rabbits (n = 20-24) of 7A at various concentrations. Incubation of lacrimocytes with 7A alone (10(-4) M), with haloperidol (10(-4) M) alone or in combination show that 7A is acting as an agonist to stimulate protein release, whereas haloperidol is acting as an antagonist to inhibit release. In vivo protein secretion rates also show a dose-response curve (at both 10 and 60 minutes post-dosing) for 7A that reach a statistically significant maximum in the dosed eye at a concentration of 0.15% w/v. Analysis of protein extracts using size exclusion HPLC shows an increase in secretory proteins, particularly tear-specific prealbumin. PMID- 8228531 TI - Theoretical corneal permeation model for ionizable drugs. AB - The primary route into the eye for many drugs is transcorneal permeation. A better understanding of the mechanisms involved in transcorneal permeation could lead to improvements in drug dosage forms or the development of drug delivery devices which enhance the ocular bioavailability of drugs. A corneal permeation model has been developed which can be used to study the mechanisms involved in corneal permeation. The model uses five compartments in series to simulate the tear film, epithelium, stroma, endothelium and aqueous humour. These tissues were assumed to be adequately represented by plane sheet barriers of physiological thickness. The tear film was assumed to be perfectly mixed and the stroma completely stagnant. Due to inadequate knowledge of the hydrodynamics of the aqueous humour, both stagnant and perfectly mixed extremes were studied. The four routes of drug loss which were considered the most significant and therefore included in the model were lacrimal drainage, conjunctival absorption, aqueous drainage and iris-ciliary body absorption. The equilibrium that can exist between the ionic and non-ionic forms of a drug was found to be an important step in the mechanism of transcorneal permeation. Including the equilibrium condition in the model resulted in aqueous humour drug levels that were over 50 times higher than the levels predicted by a model which did not use the equilibrium mechanism. A relationship between the lipophilicity of each of the two drug forms and its permeability in each layer of the cornea was used in the model. The model was used to predict aqueous humour drug concentrations resulting from a constant release of timolol into the tear film or from the application of timolol, levobunolol and pilocarpine eyedrops. The model produced transient aqueous humour drug levels that closely followed experimental in vivo data from literature. Using the model, it was also possible to predict the amount of instilled drug that is lost through each of the four elimination routes of the eye. PMID- 8228532 TI - The effects of enflurane on ocular blood flow. AB - Although general anesthesia is frequently chosen for eye surgery or experimental studies of ocular blood flow, there are few data available describing its effects on ocular blood flow. In a previous study in cats, we reported that enflurane produced significant increases in preretinal oxygen tension, indicating an increase in oxygen availability in the retina. To examine whether this effect was due to an increase in retinal or choroidal blood flow, we used radioactively labeled 15 microns microspheres of Ce 141, Sn 113, Ru 103, or Nb 95, to measure ocular blood flow in cats during enflurane anesthesia. In 10 adult cats, retinal blood flow measured 75 +/- 13, 90 +/- 9 and 88 +/- 11 ml x 100 g-1 x min-1 (mean +/- S.E.M.) at 0.5, 1.0, and 1.5 MAC enflurane, respectively (1 MAC is the concentration at which 50% of subjects do not move in response to a standardized stimulus). Corresponding values for choroidal blood flow were 1275 +/- 124, 876 +/- 106 and 842 +/- 102 ml x 100 g-1 x min-1 (mean +/- S.E.M.) at 0.5, 1.0, and 1.5 MAC enflurane, respectively. The decrease in choroidal blood flow was significant between 0.5 and 1.0 MAC. These results differ from those in our previous investigation of the effects of halothane on ocular blood flow. With halothane, retinal blood flow increased and choroidal blood flow decreased throughout the entire dose range (0.5, 1.0, and 1.5 MAC). We conclude that inhalational anesthetic agents produce significant but different effects upon ocular blood flow. PMID- 8228533 TI - Quantitative analysis of retinal vessel changes in galactose-fed dogs. AB - Retinal vessel changes in 36-month 30% galactose-fed beagles treated with or without aldose reductase inhibitors were quantified using an Olympus Cue-3 color image analysis system. Individual maps of the intact retinal vasculature, isolated by trypsin-digestion, were divided into 24 distinct subregions and measurements of either the endothelial cell to pericyte (E/P) ratio or cell densities, expressed as pericytes per mm capillary length or endothelial cells per mm capillary length, were conducted in 0.1 mm2 areas surrounding the midpoints of 12 subregions associated with the highest incidence of microaneurysms. Significantly increased E/P ratios and decreased pericyte densities were observed with the duration of galactose-feeding. These retinal changes were reduced by aldose reductase inhibitor treatment. Correlations between the E/P ratio and either number of microaneurysms or cataract severity were also observed. These data support the dose-dependent effects of aldose reductase inhibitors in preventing pericyte degeneration and subsequent formation of microaneurysms (Archives Ophthalmol. 108:1301, 1990). PMID- 8228534 TI - Continuous flow perfusion of gentamicin with a scleral shell reduces bacterial colony counts in experimental Pseudomonas keratitis. AB - We have previously shown the pharmacokinetic value of delivering gentamicin to the rabbit anterior segment using the Morgan Therapeutic Lens. The present study utilized an intrastromal injection model of Pseudomonas keratitis to test the therapeutic efficacy of continuous flow delivery of gentamicin with the Morgan therapeutic lens. All eyes (n = 52) received an intrastromal injection of approximately 1800 colony forming units (CFU) of Pseudomonas aeruginosa. At 22 hours after injection, eyes were perfused for 6 hours with saline or gentamicin (1, 2.5 or 5 mg/ml), or received gentamicin drops (13.6 mg/ml) at 15 minutes for four doses, then hourly for 6 hours. Corneas were homogenized and plated to determine bacterial survival, and expressed as log colonies (CFU). Log CFU recovered were 7.37 +/- 0.04, 6.64 +/- 0.20, 5.64 +/- 0.31, and 3.56 +/- 0.50 log CFU for saline perfusion, 1, 2.5, 5 mg/ml gentamicin perfusion respectively. Following six hours of treatment with topical fortified gentamicin drops, 5.93 +/ 0.34 log CFU were recovered. Gentamicin perfusion (5 mg/ml) was significantly different from saline or the other treatment groups (P < 0.05). Continuous corneal perfusion with the Morgan Therapeutic Lens demonstrated an increasing dose response curve with increasing perfusate concentration. It was effective in the treatment of experimental Pseudomonas keratitis. PMID- 8228535 TI - Iontophoresis of methylene blue for gonioscopic pulsed dye laser sclerostomy. AB - Iontophoresis of methylene blue using pipette tip probes is an effective means of scleral staining in preparation for gonioscopic pulsed dye laser sclerostomy. The effects of the surface area of the pipette tip and of the duration of iontophoresis on the concentration and distribution of stain were investigated in rabbit eyes. Iontophoresis of 1% methylene blue was done using currents from 0.2 to 2.0 mAmp, pipette tip probes with surface areas from 0.02 to 7.1 mm2, and durations of iontophoresis from 0.5 to 10 minutes. Optimum iontophoresis parameters for delivering methylene blue to the eye were a current of 0.4 mA, a probe tip surface area of 0.19 mm2, and a duration of five minutes. The maximal scleral stain concentration, approximately 0.4%, was achieved using these optimal parameters. Microscopic analysis of frozen tissue sections demonstrated the stain to have penetrated the full thickness of the sclera after five minutes. Tissue samples were analyzed for dye 0.5, 2, 5, 12, and 24 hours after iontophoresis was completed. Decreased dye concentration by over 50% within 2 hours and complete disappearance of dye within 24 hours were demonstrated. Methylene blue concentration thresholds for laser ablation were also examined; the threshold of ablation of methylene blue was 0.0625% when an energy level of 75 mJ was used. We propose that iontophoresis of methylene blue is a useful adjunct to pulsed dye laser sclerostomy formation. PMID- 8228536 TI - Craniocervical necrotizing fasciitis. AB - Craniocervical necrotizing fasciitis is a serious, life-threatening infection affecting fascial planes within the head and neck. Frequently the result of odontogenic origin, these polymicrobial infections, often involving group A beta hemolytic streptococcus acting in synergy with an anaerobe, can spread rapidly, causing significant morbidity and mortality, and must be diagnosed as early as possible. Treatment consists primarily of appropriate intravenous antibiotics and aggressive surgical debridement. PMID- 8228537 TI - Radiosurgery for acoustic neuroma. AB - Radiosurgery has been used for many years with great success for the treatment of acoustic neuroma. We present three cases in Louisiana using the E L Fisher system, the results of treatment, and a discussion of acoustic neuroma. PMID- 8228538 TI - Early prenatal diagnosis: the first 100 cases of chorionic villus sampling at Ochsner Foundation Hospital. AB - Chorionic villus sampling is an exciting addition to prenatal diagnosis. With this procedure, accomplished quickly and with minimal discomfort at 9 to 11 weeks of pregnancy, information can be obtained about the fetal chromosomal complement. In addition, the fetus can be tested for a variety of disorders, such as sickle cell disease, Tay-Sachs, and cystic fibrosis. The majority of fetal studies are normal, and such early prenatal testing provides relief of anxiety and opportunity for early maternal-fetal bonding. PMID- 8228539 TI - Response of automatic atrial tachycardia to intravenous adenosine. AB - The electrophysiologic responses to intravenous adenosine observed in a patient with automatic atrial tachycardia are presented. Although the development of advanced AV block was not unexpected, prolongation of atrial cycle lengths before the abrupt termination of the arrhythmia, and the subsequent transient, but total, suppression of all electrical cardiac activity were. The atrial arrhythmia reappeared when adenosine effects dissipated. The current tendency to use intravenous adenosine as the drug of first choice in the acute management and diagnosis of paroxysmal supraventricular arrhythmias, despite its lack of superiority over verapamil, will probably result in many more reports of unexpected adverse effects following its use. PMID- 8228540 TI - Health care for the homeless: a public concern. AB - The number of homeless people in the nation and New Orleans continues to increase. It is an important problem for all citizens but is especially so for the social and medical care agencies. As physicians we are morally bound to help the less fortunate of our communities, and we must also protect the community from the illnesses to which the homeless may be more susceptible. However, because very little is accurately and reliably known about the homeless in New Orleans, little can be efficiently done to aid them. We report the results of our brief surveys to stimulate concern and action in the medical community for these unfortunate people. PMID- 8228541 TI - ECG of the month. In sickness and in health. Atrial flutter. PMID- 8228542 TI - Herpes zoster oticus: diagnosis and management. AB - Herpes zoster oticus (Ramsay Hunt syndrome) is recognized as a polycranial neuritis caused by the DNA virus Herpes zoster and characterized by damage to sensory and motor nerves, including the audio-vestibular apparatus. Common presenting symptoms include cutaneous auricular vesicles, severe otalgia, inflammation of the pinna, and occasionally unilateral sudden facial paralysis. This article reviews the medical management of this disease, including the efficacy of antibiotics, corticosteroids, and acyclovir, along with the role of surgical decompression of the facial nerve. PMID- 8228543 TI - Ethical considerations in the management of terminally ill patients. PMID- 8228544 TI - Adolescent suicide. PMID- 8228545 TI - Cholesterol levels in 1,084 healthy New Orleans males. AB - It is widely recognized that elevated cholesterol levels constitute a major risk factor for the development of atherosclerosis and coronary heart disease. Most of the previous surveys conducted in an effort to learn more about incidence of hypercholesterolemia involved patients who had other concurrent risk factors such as hypertension, obesity, diabetes mellitus, cigarette smoking, or a history of myocardial infarction. Relatively few studies have been conducted in younger populations or in healthy individuals. Because we had access to baseline cholesterol data on 1,084 relatively young, otherwise completely healthy, nonobese males, we elected to determine the prevalence of hypercholesterolemia in this population. Elevated cholesterol levels (> 200 mg/dL) were found in 25.2% of our healthy subjects. These findings help to confirm the presence of a potentially serious public health problem existing among otherwise healthy, relatively young men in our community. PMID- 8228546 TI - Male breast cancer. AB - Male breast cancer is a rare tumor consisting only 1% of all breast cancers. Although 90% of breast masses in men are malignant, delay in diagnosis often occurs with resultant advanced local or systemic disease common. The few clinical studies on male breast cancer suggest that the disease behaves similarly to female breast cancer. Management, therefore, should be similar to the management of female breast cancer. Stage I, II, and III disease require a multimodal approach with regional therapy, adjuvant chemotherapy, and hormonal therapy. Most male breast cancers are estrogen or progesterone receptor positive and responses to hormonal therapy are high. A trial of hormonal therapy is warranted in most patients with stage IV disease. PMID- 8228548 TI - Futures in your future? PMID- 8228547 TI - The Family Practice Residency in Baton Rouge: practice patterns of graduates. AB - One hundred twenty-four physicians have completed the Baton Rouge Family Practice Residency since 1973. Eighty-two percent of the graduates who are currently active physicians are practicing medicine in Louisiana. Most are practicing family medicine. The graduates were sent questionnaires asking about practice characteristics, office procedures performed, hospital privileges held, and career satisfaction. Responses were received from 74 family physicians serving small, medium, and large communities. Most reported they are satisfied with their career choices. The respondents perform a variety of procedures in their offices. This finding was not affected by community size. Hospital privileges were affected by community size. The residency is meeting its principal mission: (1) training family practitioners for Louisiana, and (2) training doctors to practice in any sized community. PMID- 8228549 TI - Risks in sickle cell trait. PMID- 8228550 TI - Molecular engineering of the pancreatic beta-cell. PMID- 8228551 TI - New anticoagulant strategies. AB - The limitations of standard heparin have prompted the development of a variety of newer antithrombotic agents. In fact, a LMWH preparation has recently been approved for clinical use in North America. Of these novel preparations, LMWH, the direct thrombin inhibitors, and inhibitors of GPIIb-IIIa have been used clinically and are in advanced stages of evaluation. Not only is LMWH effective in the prevention of venous thromboembolic disease in high-risk patients, but its more predictable dose response makes it an ideal candidate for the treatment of venous thrombosis. Further studies are needed to determine whether LMWH is superior to standard heparin as adjunctive therapy in patients undergoing coronary thrombolysis or angioplasty. Particularly promising in the setting of arterial thrombosis are hirudin, hirulog, and 7E3. With the encouraging results reported to date, it is likely that these agents will soon find their way into the treatment armamentarium of arterial thrombosis. PMID- 8228552 TI - Cardiopulmonary function in men with sickle cell trait who reside at moderately high altitude. AB - Sickle cell trait has been associated with an unexplained increased risk of sudden death during military basic training. Previous studies of cardiopulmonary function in persons with sickle cell trait at sea level or after brief exposure to moderately high altitude have not shown significant abnormalities. To determine whether cardiopulmonary function is impaired in men with sickle cell trait who chronically reside at moderately high altitude, we prospectively studied 17 men with sickle cell trait and 25 men with normal hemoglobin. All subjects had resided at > or = 1609 m above sea level for at least 10 years. Resting pulmonary function and cardiopulmonary performance during exercise to exhaustion were evaluated. No statistically significant differences were observed between subjects with sickle cell trait and controls for any of the resting pulmonary function variables measured. All parameters of exercise performance were not statistically different between the groups. Increasing age was associated with a similar decrease in work capacity and oxygen consumption in both groups. Electrocardiogram findings were not statistically different between groups. We conclude that chronic exposure to moderately high altitude does not impair cardiopulmonary function in men with sickle cell trait. PMID- 8228553 TI - Angiotensin-converting enzyme and myocardial fibrosis in the rat receiving angiotensin II or aldosterone. AB - Angiotensin-converting enzyme (ACE) is present in tissues composed largely of fibrillar collagen such as heart valves, the adventitia of great vessels and intramyocardial coronary arteries, and the scar that follows myocardial infarction. We tested the hypothesis that such tissue ACE is related to fibrous tissue formation and that its appearance is independent of circulating renin and angiotensin peptides. For this purpose we selected experimental models that simulate primary and secondary hyperaldosteronism, each of which are associated with the appearance of myocardial fibrosis. ACE in the excised heart and aorta was localized by in vitro autoradiography with [125I]351A, while fibrosis was identified by light microscopy in sections stained with collagen specific picrosirius red. Tissue was obtained at 2, 4, and 6 weeks from various experimental groups: unoperated, untreated, age- and sex-matched controls; age- and sex-matched uninephrectomized control rats on a high sodium diet; and rats that had received either aldosterone (ALDO) or angiotensin II (AII). Compared with controls, we found ACE binding (1) unchanged after 2 weeks of ALDO, but increased in the adventitia of intramural coronary arteries after 4 weeks, in keeping with the perivascular fibrosis that appeared in each ventricle; (2) markedly increased after 6 weeks of ALDO, where it not only involved coronary vessels but also microscopic scars that appeared in atria and ventricles; (3) increased in the coronary adventitia and sites of scarring, each of which were present 2 weeks after AII; and (4) markedly increased after 4 and 6 weeks of AII as fibrosis became more extensive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228554 TI - Angiotensin II and aldosterone receptor binding in rat heart and kidney: response to chronic angiotensin II or aldosterone administration. AB - Angiotensin II (AII) and mineralocorticoid receptors (MRs) have been identified in the mammalian heart and kidney. Their response to chronic elevations in circulating AII or aldosterone (ALDO) (or both) is unknown in either primary or secondary hyperaldosteronism. Nonendothelial angiotensin-converting enzyme (ACE) activity has been found in the fibrous tissue response that involves intramyocardial coronary arteries and microscopic scarring. Whether this tissue ACE could affect AII receptors and MRs in the area of myocardial fibrosis is likewise unknown. To address these questions, we monitored AII and MR binding in the rat heart and kidney after chronic AII or ALDO infusion. All receptor and MR binding were localized by in vitro autoradiography with 125I-[Sar1, Ile8]-labeled AII and [1,2,6,7 3H]-labeled ALDO, respectively. To characterize AII receptor subtypes, a type I antagonist (DuP753) or type II antagonist (PD 123177) was used. Four experimental groups were examined: unoperated, untreated, age- and sex matched controls; age- and sex-matched uninephrectomized control rats receiving a high sodium diet; animals that received AII (9 micrograms/hr sc) for 2, 4, 6, or 8 weeks; and uninephrectomized rats on a high sodium diet that received ALDO (0.75 microgram/hr sc) for similar periods of time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228555 TI - Endogenous testosterone, fibrinolysis, and coronary heart disease risk in hyperlipidemic men. AB - To assess relationships of endogenous testosterone with fibrinolysis and coronary heart disease risk factors in 55 newly referred hyperlipidemic men, we studied the relationships of testosterone to basal fibrinolytic activity, lipids, lipoproteins, and apolipoproteins. Testosterone correlated positively with the major stimulator of fibrinolysis, tissue plasminogen activator activity (r = 0.30; p = 0.02) and correlated inversely with two independent coronary heart disease risk factors, plasminogen activator inhibitor activity, the major fibrinolysis inhibitor (r = -0.33; p = 0.01), and fibrinogen (r = -0.39; p = 0.004). Testosterone correlated inversely with plasma triglycerides (r = -0.33; p = 0.01). Stepwise multiple regression was done with fibrinolytic activities as the dependent variables, and age, Quetelet Index (relative ponderosity), apolipoprotein A-I, apolipoprotein B, triglyceride, testosterone, time of blood sampling, and lipoprotein (a) as explanatory variables. Testosterone was an inverse, independent predictor of fibrinogen (p = 0.002); 53% of the variance of fibrinogen could be accounted for by age and triglyceride level (positive; p = 0.001, p = 0.01), and by apolipoprotein A-I and testosterone (negative; p = 0.02, p = 0.002). Testosterone was an independent inverse predictor of tissue plasminogen activator antigen (p = 0.0008), with tissue plasminogen activator antigen correlating inversely with tissue plasminogen activator activity. Quetelet index and apolipoprotein B were independent negative predictors of tissue plasminogen activator activity (p = 0.02, p = 0.03); Quetelet index and triglycerides were independent positive predictors of plasminogen activator inhibitor activity (p = .0001, p = .0001) and alpha 2-antiplasmin (p = 0.0003, p = 0.009).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228556 TI - Regulation of hepatic insulin receptor tyrosine kinase in rat models of mild insulin resistance. AB - Hepatic insulin receptor tyrosine kinase activity (IR-TKA) correlates with plasma insulin levels in severely hyperinsulinemic Zucker rats (Diabetes 1990; 39: 619 625). We tested such a correlation in rat models of insulin resistance with plasma insulin concentrations in the physiologic range. Female Sprague Dawley rats drank either 10% glucose (G; n = 7) or 10% fructose (F; n = 5) in their water for two months and were compared to a control group (C; n = 6). The rats in both experimental groups developed hyperglycemia (G = 8.4 +/- 0.3, F = 8.5 +/- 1.1, C = 4.9 +/- 0.3 mM) but plasma insulin levels did not differ significantly (G = 144 +/- 30, F = 162 +/- 12, C = 108 +/- 6 pmol/L). There were no differences observed in any function of the hepatic insulin receptors between the rat groups. 125I-insulin binding to purified liver receptors was normal. However, for all groups, there was a significant correlation between in vivo plasma insulin levels and IR-TKA measured both in the absence (r = 0.47, p < 0.05) and presence (r = 0.57, p < 0.02) of added insulin in in vitro experiments. The same correlation was seen for the insulin/glucose ratio (as a measure of in vivo insulin resistance) vs. IR-TKA (r = 0.62, p < 0.01; r = 0.49, p < 0.05, in the basal and insulin-stimulated states, respectively). Thus, insulin in the physiologic range regulates TKA of rat liver IR. The TKA of hepatic IR isolated from such rats maintains the ability to be further stimulated by exogenous insulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228557 TI - Do patients with antiphospholipid syndrome have autoantibodies to beta 2 glycoprotein I? AB - High positive anticardiolipin antibody tests have been associated with recurrent thrombosis and pregnancy loss. Although these antibodies were believed to bind negatively charged phospholipids, recent reports have suggested that a serum protein, beta 2-glycoprotein I (beta 2-GPI), may be the true antigen for these antibodies. To resolve this issue, we compared binding of 75 anticardiolipin positive and 71 anticardiolipin-negative serum samples from patients with rheumatic diseases to beta 2-GPI by using an enzyme-linked immunosorbent assay (ELISA). Serum samples from 30 healthy blood donors and 10 laboratory personnel were used as normal controls. We found no difference in binding between the three groups of serum samples. In addition, when binding to beta 2-GPI coated plates was compared with binding to ELISA plates without beta 2-GPI (blank), no difference was observed. Finally, binding of anticardiolipin-positive serum samples to plates coated with cardiolipin-beta 2-GPI mixture varied directly with the cardiolipin concentrations. Based on these findings, we conclude that anticardiolipin-positive serum samples do not bind beta 2-GPI. PMID- 8228558 TI - Effects of magnesium on the renin-angiotensin-aldosterone system in human subjects. AB - Recent prospective study of intravenous magnesium sulphate administration into patients with acute myocardial infarction revealed the improvement of survival rate of these patients. This study was designed to determine the effect of intravenous administration of magnesium on the renin-angiotensin-aldosterone (RAA) system. MgSO4 (0.72% in 3.24% glucose solution) at 5 mEq/hr was infused intravenously into six healthy normotensive men for six hours. MgSO4 infusion increased plasma renin activity (PRA) at 60 minutes (4.1 +/- 1.4 ng/ml/hr vs 2.7 +/- 0.8 ng/ml/hr, p < 0.05) and decreased plasma aldosterone concentration (PAC) at 180 minutes (40 +/- 8 pg/ml vs 60 +/- 12 pg/ml, p < 0.05). The maximum level of serum magnesium reached 2.32 +/- 0.09 mEq/L, from the control value of 1.58 +/ 0.07 mEq/L, at -3 hours. Analysis of both serum levels and urinary excretion of electrolytes revealed significant increases in urinary excretion of calcium and magnesium, whereas no significant changes in other electrolytes in serum or urinary excretion were found. To clarify the mechanism by which magnesium increased PRA and suppressed PAC, either prostaglandin synthesis inhibitor, indomethacin (75 mg/day), or a calcium channel blocker, diltiazem (90 mg/day), was administered. Indomethacin completely blocked the increase in PRA but did not induce any changes in PAC. Diltiazem inhibited the decrease in PAC but did not induce any changes in PRA. These data indicate that magnesium stimulates renin release through the elevation of prostaglandins and suppresses aldosterone production through the intracellular calcium mobilization. From these results it is suggested that a favorable effect of magnesium administration to patients with myocardial infarction relates to the alterations of the RAA system. PMID- 8228559 TI - Failure to mobilize intracellular calcium in response to thrombin in a patient with familial thrombocytopathy characterized by macrothrombocytopenia and abnormal platelet membrane complexes. AB - We report a mother and son who were found to have macrothrombocytopenia, prolonged bleeding time, and abnormal platelet responses to thrombin. Transmission electron microscopy performed on the son's platelets demonstrated an unusual arrangement of membrane complexes formed by association of the open canalicular and dense tubular systems. Number and appearance of platelet alpha granules, dense bodies, and mitochondria were normal. These platelets demonstrated normal agonist-induced Ca2+ flux in response to collagen and supranormal responses to arachidonic acid but displayed no increase in intracellular free Ca2+ in response to thrombin. Platelet surface glycoproteins IIb-IIIa, Ib, and granular membrane protein-140 measured by fluorescence activated flow cytometry, along with platelet content of von Willebrand factor and fibrinogen, were normal. The von Willebrand factor binding function of GP-Ib on these platelets was also normal. We believe that this family demonstrates a unique macrothrombocytopenia syndrome characterized by deficient Ca2+ mobilization in response to thrombin that is not related to a defect in GP-Ib. PMID- 8228560 TI - Octreotide and cholecystokinin antagonist reduce edema in obstruction-induced acute pancreatitis. AB - Obstruction-induced acute pancreatitis in rats is associated with increased plasma cholecystokinin (CCK) levels. Duodenal replacement of bile reduces severity of pancreatitis and limits CCK increase. We investigated the role of CCK in the pathogenesis of obstruction-induced acute pancreatitis by pretreating rats with the somatostatin analog octreotide and the CCK antagonist L-364,718. Octreotide inhibits duodenal CCK release, and L-364,718 competitively blocks CCK receptors. We studied 31 rats after (1) sham operation (n = 7), (2) bile and pancreatic duct obstruction (BPDO) (n = 12), (3) BPDO plus octreotide (20 micrograms/kg IP and then 5 micrograms/kg/hr IV) (n = 6), and (4) BPDO plus L 364,718 (1 mg/kg IP and then 0.25 mg/kg/hr IV) (n = 6). Rats were killed after 18 hours. Pancreas weight, acute pancreatitis histology score, and plasma amylase and CCK levels were determined. Octreotide and L-364,718 limited the increase in pancreas weight. Octreotide also limited the rise in plasma CCK levels. These findings suggest that CCK may play a role in the pathogenesis of obstruction induced acute pancreatitis. PMID- 8228561 TI - Influence of heat on platelet biochemistry, structure, and function. AB - The present investigation has evaluated the influence of temperatures ranging from 37 degrees C to 45 degrees C for intervals of 30, 60, and 90 minutes on the biochemistry, morphology, and function of human platelets. Exposure to temperatures up to 43 degrees C for an hour did not significantly alter platelet morphology or physiologic response to aggregating agents. Samples of platelets heated at 43 degrees for 60 minutes lost their ability to aggregate in response to arachidonate, but sensitivity was restored by pretreatment with epinephrine. Platelets heated to 45 degrees C for 90 minutes were converted from discs to spheres and failed to aggregate in response to all agonists, whether or not they were pretreated with epinephrine. The platelets heated at 45 degrees C for 90 minutes could adhere to formvar or denuded subendothelium but were unable to extend pseudopods or to spread. They maintained normal levels of adenine nucleotides and serotonin but failed to secrete these products on stimulation. Studies with fibrinogen coupled to gold (Fgn/Au) revealed only a few particles bound to glycoprotein IIb-IIIa (GPIIb-IIIa) on platelets heated at 43 degrees C for 90 minutes. Ligands that did bind to GPIIb-IIIa were transported to the open canalicular system. Biochemical studies demonstrated normal synthesis of thromboxane B2 and calcium flux by platelets heated at 45 degrees C for 90 minutes. Polyacrylamide gels of platelets heated at 45 degrees C for 90 minutes showed an increase in talin incorporation into heated platelet cytoskeletons but no increase in filamentous actin. The findings indicate that impaired function of heated platelets is due to the influence of heat on cytoskeletal proteins important for pseudopod extension, shape change, expression of GPIIb-IIIa, or other surface membrane receptors and secretion, but that impaired function is not due to inhibition of biochemical systems involved in activation events. PMID- 8228562 TI - Enhanced expression of mRNA for transforming growth factor-beta, type I and type III procollagen in human post-burn hypertrophic scar tissues. AB - To explore the possible role of locally synthesized transforming growth factor beta-1 (TGF-beta 1) and procollagen gene expression in postburn hypertrophic scars, we have compared mRNA levels for type I and type III procollagen and TGF beta 1 in human hypertrophic scar tissue with normal dermis obtained from the same patients as a control. Northern blot analysis of total RNA extracted from hypertrophic scar tissue and normal skin demonstrated two transcripts for the pro alpha 1(I) chain (5.8 kb and 4.8 kb) and for the pro-alpha 1(III) chain (5.4 kb and 4.8 kb) and one transcript (4.9 kb) for TGF-beta 1. Quantitative analysis of dot blot autoradiograms of mRNA from three samples of hypertrophic scar tissue and normal skin showed average increases of 102% (p < 0.05) for pro-alpha 1(I), 91% (p < 0.06) for pro-alpha 1(III), and 61% (p < 0.05) for TGF-beta 1. Three additional hypertrophic scar samples were quantitatively analyzed on Northern blots and showed increases of 246%, 102%, and 250% of the specific messages for pro-alpha 1(I), pro-alpha 1(III), and TGF-beta 1 relative to a normal skin control. Two transcripts (4.9 kb and 2.5 kb) for TGF-beta 1 were identified in cultured fibroblasts. In contrast to the results from tissue, the level of these transcripts in fibroblasts cultured from hypertrophic scar tissue and normal skin were not significantly different, suggesting that the synthesis of this growth factor is stimulated in tissue by a presently unknown mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228563 TI - Ozone in all its reactive splendor. PMID- 8228564 TI - The reticuloendothelial system and bloodstream clearance. PMID- 8228565 TI - Eicosanoids and human colon cancer. PMID- 8228566 TI - The third component of complement: new functions for an old friend. AB - The present review has evaluated four biologic processes in which the participation of C3 as a primary mediator deserves more extensive investigation. These novel functions of C3 derive in part from the biochemistry ascribable to a thiolester-containing protein and are underscored by its remarkably high concentration in plasma and widespread presence in tissues. Such physiologic characteristics argue that we have only begun to understand the functions of this most versatile complement protein. The availability of full-length cDNA and the ability to manipulate the C3 gene in tissue culture or embryonic stem cells should help to illuminate the pluripotent nature of this fascinating protein. PMID- 8228567 TI - Oxidation of biologic molecules by ozone: the effect of pH. AB - Ozone (O3) is a powerfully oxidizing pollutant gas. Its toxic effects to animals appear to be worsened by coexposures to acid-generating compounds such as oxides of nitrogen and sulfur. Ozone (16 ppm) oxidizes ascorbic acid and uric acid (two important antioxidants in lung lining fluids) at equal rates at pH 5.0 or pH 7.4. Loss of intrinsic fluorescence and formation of carbonyls in albumin exposed to O3 are similar at both pH values. However, albumin-SH groups are lost much faster on exposure to O3 at pH 7.4 than at acidic pH values. A similar slower rate of SH group disappearance at acidic pH is seen when cysteine or reduced glutathione are exposed to O3. We suggest that the ability of reduced glutathione, albumin, and other proteins containing -SH groups to scavenge O3 in the respiratory tract is impaired at low pH and that this effect could contribute to the aggravation of O3 toxicity. PMID- 8228568 TI - Effect of ethanol on immune clearance in mice: biphasic alteration of complement mediated clearance with chronic ethanol ingestion. AB - Rate constants (k1 through k4) describing complement-mediated and Fc gamma receptor-mediated components of immune clearance were serially determined in BALB/c mice fed ethanol, 10%, in drinking water, for 24 weeks. A branched series, first order reaction sequence model of immune clearance was used to obtain the rate constants from measurements of the clearance of radiolabeled immunoglobulin G-opsonized, murine erythrocytes. A > 50% decrease in complement-mediated clearance occurred, with a nadir of complement-mediated sequestration (k1) and complement-dependent phagocytosis (k4) at 2 weeks (p < 0.003). Mean k1 and k4 rate constant values returned to control levels by week 6, and k1 increased to elevated values in weeks 10 through 20 (p < 0.05). The rate constant governing C3b deactivation and return of deactivated, sensitized cells back to the circulation (k2) was initially normal but decreased in weeks 6 through 24 (p < 0.05). Neither immunoglobulin G Fc gamma receptor-mediated clearance nor the survival of nonsensitized cells were decreased by ethanol. Mice fed ethanol had a mean blood alcohol level of 14.9 +/- 7.2 mmol/L, and their mean weight and serum complement levels did not differ from untreated controls. Complement-dependent sequestration and phagocytosis did not decrease significantly when rechallenged with 10% ethanol, but the decrease in k2 and increase in k1 did occur on rechallenge. Thus, chronic ethanol ingestion in mice is associated with an initial decrease followed by a small rebound increase in complement-mediated clearance of opsonized cells. Fc gamma receptor-mediated clearance is not decreased, and only the rebound increase in complement-mediated clearance is observed on rechallenge. This model provides a unique opportunity to study selective in vivo effects of ethanol on an important function of the immune system as well as to explore the mechanisms of ethanol tolerance in mice. PMID- 8228569 TI - Altered eicosanoid levels in human colon cancer. AB - Eicosanoids may participate in colon carcinogenesis, as evidenced from work in animal tumor models showing prevention of colon cancer by inhibitors of their synthesis and epidemiologic studies demonstrating reduced risk of colon cancer in long-term users of aspirin and other nonsteroidal antiinflammatory drugs (NSAIDs). The levels of prostaglandin E2 (PGE2), PGF2 alpha, PGI2, thromboxane A2 (TXA2), and leukotriene B4 (LTB4), which represent the cyclooxygenase and 5 lipoxygenase pathways, were determined in 21 pairs of surgically excised human colon cancer and histologically normal mucosa samples 5 to 10 cm away from the tumor. The levels of PGE2 were elevated in colon cancer samples as compared with histologically normal mucosa samples distant from the cancer (p < 0.01), whereas levels of prostacyclin (PGI2) were decreased (p < 0.05). The differences in the levels of PGF2 alpha, TXA2, and LTB4 between normal and malignant tissue were not statistically significant. No statistically significant association was found between the level of each of the eicosanoids assayed and Dukes' stage of colon cancer. These findings, confirming and extending earlier work from tumors and cell culture, suggest that the protective effect of aspirin and other NSAIDs in the development of human colon cancer may be mediated, at least in part, through their inhibition of arachidonic acid metabolism by cyclooxygenase. PMID- 8228570 TI - Lung structure and elastic recoil properties in hereditary diabetes mellitus in KK-mice, C57 black mice, and F1 hybrids. AB - To separate the effects of diabetes and inheritance on morphometric characteristics of the lung and on the variables of the pressure-volume relationship, the lungs of KK mice--a genetic model of adult-onset diabetes--were compared with the lungs of normal C57 black mice and of F1 hybrids by multivariate analysis. Hybrid mouse lungs contain more air per gram of lung tissue at total lung capacity (TLC) than the lungs of other mice (p < 0.0002) and have a lower recoil pressure at 30% to 70% of TLC. Morphometric data revealed that both the mean linear intercept and the number of alveoli per unit volume in hybrid mice fell between those in KK and C57 mice. However, the total number of alveoli per lung in hybrid mice was 155% of that in KK mice and 148% of that in C57 mice, reflecting the larger lung volume of hybrid mice. The alveolar wall thickness of the hybrids was similar to that of C57 mice, whereas KK mice had thicker alveolar walls than the other two groups. Multivariate analysis revealed that body weight, wet lung weight, volume density of blood vessels, mean linear intercept, and transpulmonary pressure at 90% TLC were affected by diabetes alone; however, the following parameters were affected by both inheritance and diabetes: lung volume, specific lung volume, femur length, volume density of alveolar air, surface area, surface to volume ratio, number of alveoli per lung, lung air per gram of lung tissue, and transpulmonary pressure at 20% to 80% TLC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228571 TI - Developmental changes of protein kinase C and Gs alpha in hypertrophic cardiomyopathic hamster hearts. AB - Protein kinase C (PKC) and GTP-binding proteins (G-proteins) are known to be major determinants in the modulation of cardiac function. In this study, we examined the developmental changes of PKC and the alpha-subunit of the stimulatory guanosine triphosphate binding protein (Gs alpha) in hypertrophic cardiomyopathic Syrian hamster (BIO 14.6) hearts, before the onset of hypertrophy (30 days old) and at the peak of hypertrophy (6 months old) and compared these with age-matched control hamster (BIO RB) hearts. At 30 days, cardiac PKC activity was similar between BIO 14.6 and BIO RB both in the membrane (117.1 +/- 9.9 pmol/min/mg vs 131.2 +/- 12.7 pmol/min/mg in controls, n = 8) and in the cytosolic fractions (213.1 +/- 22.0 pmol/min/mg vs 186.6 +/- 23.9 pmol/min/mg in controls, n = 9). At 6 months, PKC activity was significantly higher in BIO 14.6 than in controls, both in the cardiac membrane (131.9 +/- 7.1 pmol/min/mg vs 40.7 +/- 4.7 pmol/min/mg in controls, n = 8, p < 0.00001) and cytosol (77.9 +/- 2.1 pmol/min/mg vs 54.6 +/- 3.3 pmol/min/mg in controls, n = 6, p < 0.0005). In BIO RB hearts, membrane and cytosolic PKC activities were significantly reduced at 6 months compared with those at 30 days of age (p < 0.001). However, the membrane PKC activity in 6-month-old BIO 14.6 was maintained at the level of the 30-day old hearts. On the other hand, the relative immunoreactive amounts of Gs alpha were similar between BIO RB and BIO 14.6 hearts at 30 days and at 6 months of age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228572 TI - Quantitative analysis of vascular prostaglandin H synthase mRNA levels by competitive polymerase chain reaction. AB - Prostaglandin H synthase (PGHS) occupies a central position in the biosynthesis of prostaglandins, thromboxane, and prostacyclin. Expression of this enzyme is enhanced by inflammatory cytokines and phorbol esters. Because of the low abundance of PGHS, PGHS mRNA levels in tissues and cells were difficult to measure accurately. In this study, we quantified vascular PGHS-1 mRNA levels by a competitive polymerase chain reaction assay. The mean basal PGHS-1 level in cultured human umbilical vein endothelial cells (HUVECs) was 24.3 +/- 10.6 amol/microgram RNA. Treatment of the HUVECs with recombinant human interleukin-1 beta at 37 degrees C for 4 hours increased the mRNA to 49.1 +/- 9.1 amol/microgram RNA, a 2.1-fold increase. Treatment of the cells with phorbol 12 myristate 13-acetate at 37 degrees C for 4 hours caused a 1.7-fold increase in the mRNA level. This procedure was utilized to measure PGHS-1 mRNA levels in rabbit abdominal aorta, inferior vena cava, liver, kidney, and lung. The aortic mRNA level (9.2 +/- 1.9 amol/microgram RNA) was about 5 times higher than the inferior vena caval level (2.4 +/- 1.3 amol/microgram RNA) but only slightly higher than the level of other organs. These results reveal a significant difference in PGHS-1 mRNA levels between arterial and venous tissues. The synthase mRNA levels in organs such as liver, kidneys, and lungs do not differ significantly. The PCR assay should be valuable for studying the role of PGHS expression in disease states. PMID- 8228573 TI - Homeostasis of zinc in marginal human zinc deficiency: role of absorption and endogenous excretion of zinc. AB - Although biochemical and immunologic aspects of mild or marginal human zinc deficiency have been well characterized, there is a paucity of data regarding the effects of prolonged marginal zinc deficiency on zinc homeostasis. It appears that human beings are able to maintain zinc homeostasis by increasing efficiency of zinc absorption and decreasing endogenous excretion of zinc when they are subjected to short-term dietary zinc restriction. However, a mild deficiency of zinc in human beings under usual circumstances is an outcome of chronic exposure to diets low in zinc lasting for many months and years. Therefore, it is important to determine whether or not the adapted zinc homeostasis during the short duration of dietary zinc deprivation is also maintained during a prolonged period of dietary zinc restriction. We assessed the efficiency of zinc absorption as well as endogenous zinc excretion during a 6-month period of dietary zinc restriction (63.1 mumol/day) in human volunteers by using a stable zinc (70Zn). Prolonged marginal zinc deficiency did not impair the functional role of endogenous zinc excretion in zinc homeostasis, but efficiency of zinc absorption was not sustained and decreased in the majority of our volunteers when the zinc restricted diet was continued for 6 months. Such altered mechanisms of zinc homeostasis suggest that chronic human zinc deficiency warrants reassessment of dietary zinc requirement to preserve normal zinc status in the subsets of population that are vulnerable to zinc deficiency. PMID- 8228574 TI - Flow cytometric analysis of residual white blood cell concentration and platelet activation antigens in double filtered platelet concentrates. AB - Reduction of contaminating leukocytes in platelet products by filtration has been shown to decrease the incidence of human leukocyte antigen (HLA) alloimmunization. Nonetheless, prevention is not complete when using current techniques, and a significant number of patients continue to exhibit clinical refractoriness and to produce alloantibodies. Interest in preventing HLA alloimmunization and other complications of white blood cell (WBC) contamination of transfused cellular products has resulted in ongoing efforts to increase the efficiency of leukodepletion filters. As the efficiency of these filters increases, more accurate and precise methods for counting extremely low numbers of WBCs must be instituted to ensure quality control. We have validated a simple, rapid flow cytometric assay for quantitating low numbers of WBCs in platelet products. The assay is sensitive to a lower limit of 0.1 WBC/microliter without concentration of the platelet product sample and has an excellent correlation (R2 = 1.00) between calculated and expected WBC concentration over a range of 0.1 to 100.0 WBC/microliter. (R2 values over the concentration ranges of 0.1 to 1.0 WBC/microliter and 1.0 to 10.0 WBC/microliter were 0.988 and 0.996, respectively.) The intraassay coefficients of variation at WBC concentrations of 50.4/microliter, 0.9/microliter, and 0.1/microliter were 4%, 8%, and 18%, respectively. The flow cytometric counting technique was applied, in concert with a Nageotte chamber manual counting method, to the enumeration of residual WBCs in 20 apheresis and random donor platelet concentrates filtered through two leukodepletion filters sterile docked in series. A greater than four log10 WBC reduction capability was demonstrated when utilizing this double filtration procedure, and its clinical applicability is underscored by data that showed no statistically significant change in expression of activation-specific platelet antigens before versus after filtration. PMID- 8228575 TI - Interleukin-4 induces tissue eosinophilia in mice: correlation with its in vitro capacity to stimulate the endothelial cell-dependent selective transmigration of human eosinophils. AB - Interleukin-4 (IL-4) has been shown to play a crucial role in the pathogenesis of allergic disease. In this study, intraperitoneal administration of IL-4 in mice led to selective accumulation of eosinophils, and intradermal injection induced marked eosinophil infiltration. However, IL-4 had no detectable effect on neutrophil accumulation. Preincubation of mouse IL-4 with the neutralizing mAb 11B11 abolished this peritoneal and dermal eosinophilia. These in vivo data correlate with the in vitro capacity of IL-4 to specifically promote the selective transendothelial migration of eosinophils. Supernatants of antigen stimulated T cell clones derived from hypersensitized individuals induced significant eosinophil transmigration that was inhibited by the neutralizing mAb 8F12 against human IL-4. These experiments impressively demonstrate a link between specific antigenic recognition and the selective recruitment of eosinophils by the endothelial barrier. Furthermore, data are presented supporting our previous evidence that eosinophils need initial priming to transmigrate across IL-4-activated monolayers. Whereas freshly isolated eosinophils from nonallergic individuals failed to transmigrate, the eosinophils from a group of patients with allergic asthma showed spontaneous layer penetration. These data further support the evidence that eosinophils from allergic patients undergo in vivo priming and are functionally different with respect to their capacity to transmigrate. PMID- 8228576 TI - Effect of ganciclovir on murine cytomegalovirus-induced reactivation of toxoplasma pneumonia. AB - Active cytomegalovirus (CMV) infection induces immunosuppression and predisposes to the development of life-threatening superinfections in immunocompromised patients. We have described a mouse model in which acute murine CMV (MCMV) infection reactivates previously dormant Toxoplasma gondii infection, manifested as pneumonia. To determine whether therapy with ganciclovir might prevent MCMV induced reactivation of T. gondii pneumonia, we administered ganciclovir to mice starting 1 day before to 2 days after MCMV infection and continuing for 14 days. When ganciclovir was begun early (before, on the day of, or 1 day after MCMV infection), the severity of T. gondii pneumonia reactivated by MCMV was significantly reduced. However, when therapy was delayed, no beneficial effect of ganciclovir was observed and the severity of MCMV-induced reactivation of T. gondii pneumonia was comparable to that seen in untreated animals. We conclude that ganciclovir therapy can attenuate but not eliminate MCMV-induced reactivation of T. gondii pneumonia and that prophylactic or very early administration of the drug is necessary to achieve protection. PMID- 8228577 TI - Membrane rigidity of red blood cells parasitized by different strains of Plasmodium falciparum. AB - Changes in the structure of parasitized red blood cells may influence their ability to circulate. We have used a micropipette technique to examine the effects of invasion and maturation of Plasmodium falciparum on the membrane rigidity of red blood cells. In the presence of immature, ring form parasites from different laboratory strains, membrane rigidity remained unchanged as compared with uninfected red cells. However, development of more mature pigmented trophozoites caused a marked increase in membrane rigidity. Parasites from knobless strains caused a less-pronounced increase than parasites from knob positive strains. Using closely synchronized cultures, the dependence of membrane rigidity on parasite maturation was studied in more detail for selected knob positive and knobless strains. Over a period of 12 hours, while trophozoites developed into schizonts, no further rigidification of the red cell membrane occurred. The increase in membrane rigidity, occurring with the initial development of pigmented trophozoites, may be related to insertion of neoantigens into the red cell surface or modification of native membrane proteins that also occur at this time. In contrast to others, we found no effect of parasite-culture supernatant, harvested at different stages, on the rigidity of uninfected cells exposed to it. Interstrain variation of membrane rigidity could influence pathophysiology in several ways: by promoting margination and cytoadherence of knob-positive strains in the microcirculation, by modulating clearance of parasitized cells by the reticuloendothelial system, and by influencing ischemic complications of severe falciparum malaria. PMID- 8228578 TI - Autoradiographic determination of permeation pathway of permeability probes across intestinal and tracheal epithelia. AB - Mucosal permeability studies are used to assess intestinal and respiratory barrier functions. Our ability to interpret results of permeability studies are hampered by our lack of understanding of absorptive pathways of permeability markers. The aim of this study was to visually trace the pathway of permeability probes across the small intestinal and tracheal epithelia by using electron microscopic autoradiography and cytochemistry. We saw a constant rate of mucosal to serosal permeation of intestinal permeability probes polyethylene glycol 400 (PEG-400) and mannitol across the everted small intestinal sac and of the pulmonary permeability probe bovine serum albumin (BSA) across the tracheal epithelia. Electron microscopic tracing of the permeation pathways of tritiated PEG-400 and tritiated mannitol revealed that the majority of the probes traversed the intestinal epithelium paracellularly within 1 half distance (1650 A) of the intercellular space. It is interesting that we also found a small but significant transcellular transport of permeability probes. Goblet cells also absorbed permeability probes transcellularly, but in an "all or none" fashion. Similar pathways were identified in studies utilizing the commonly used pulmonary permeability probes iodine 125-labeled BSA and horseradish peroxidase to determine the routes of transfer in the airway epithelia. In the normal unperturbed trachea, these large permeability probes traversed the pulmonary epithelia transcellularly via endocytosis. On barrier disruption by cytochalasin D, the probes permeated predominantly via the paracellular pathway. This study also demonstrates some of the similarities and differences in transmucosal pathways shared by intestinal and pulmonary epithelia. PMID- 8228579 TI - Plasma reactivity to hexagonal II phase phosphatidylethanolamine is more frequently associated with lupus anticoagulant than with antiphosphatidylethanolamine antibodies. AB - In membranes, phosphatidylethanolamine (PE) is usually in a lamellar phase, but it can also adopt a hexagonal II phase (HexII PE), which has been demonstrated to be immunogenic in mice and has recently been used in a new clotting assay to detect lupus anticoagulant (LA). A population of 58 patients susceptible of having LA or antiphospholipid antibodies (aPLA), was screened for reactivity to HexII PE using the new clotting assay and for the presence of anti-PE antibodies (aPEA) using ELISA. The patients could be classified as follows: 26 systemic lupus erythematosus, 14 primary antiphospholipid syndromes and 18 miscellaneous non autoimmune disorders. HexII PE reactivity was detected in 22 patients. It was significantly more often associated with LA-positivity (20/29) than with LA negativity (2/29), (p < 0.0001). However, 9 patients with LA had negative HexII PE assays. Anti-PE antibodies were detected by ELISA in only 12% of the cases. No correlation was observed between the distribution of aPEA and that of HexII PE reactivity. Interestingly, 2 LA- and aPLA-negative patients exhibited HexII PE reactivity. The absence of identity between the distribution of HexII PE reactivity and that of either LA or aPEA indicates that this new assay has its own specificity. It might detect antibodies to the immunogenic hexagonal PE and may therefore have a broader scope than that proposed initially. Although in the studied populations, no significant relationship could be found between HexII PE positivity and clinical complications (thrombosis and/or recurrent fetal losses), it might be associated with other complications of the antiphospholipid syndrome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228580 TI - Plasma fibrinolytic activators and their inhibitors in women suffering from early recurrent abortion of unknown etiology. AB - The fibrinolytic response to a venous occlusion test was evaluated in 116 consecutive patients who had each had early recurrent miscarriages of unknown origin. A normal response to venous occlusion was defined from the statistical analysis of data obtained in a matched group of 90 women with no past abortion. Seventy-four patients had at least one abnormal fibrinolysis-related test, whereas no member of the control group had any (p < 10(-6)). Fifty-six of the patients had a stasis-induced shortening of the euglobulin clot lysis time that was less than normal (median decrement 3.5% [normal > or = 28%] [p < 10(-6)]). In this subgroup, 17 women also had a poor tissue-type plasminogen activator (tPA) response (median increment 18% [normal > or = 34%] [p < 0.001]). Twenty-one of the 56 had high plasma levels of type 1 plasminogen activator inactivator (PAI-1) as well as a high PAI activity level (p < 0.001 for comparison with normal controls). Sixteen patients had both abnormalities (tPA response blunted and high PAI values). When retested in the same patients, the patterns persisted over time. Nine other habitual aborters had abnormally high type 2 plasminogen activator inhibitor plasma levels (> 4.2 ng/ml). Six other patients had a poor urokinase-type plasminogen activator reactivity to venous occlusion (post-stasis levels lower than 0.18 ng/ml). Thus activators and inhibitors of the fibrinolytic system are frequently abnormal in primary habitual aborters. The current pathogeneses of high PAI-1 concentrations and of impaired tPA reactivity to venous occlusion were not found in these patients. The role of the fibrinolytic components in trophoblast invasiveness has been suspected in animals, and an impaired plasmin-dependent proteolysis in women might favor recurrent abortion by promoting fibrin deposition in early placental circulation or by limiting trophoblast development, or both. PMID- 8228581 TI - Factors influencing the use of primary care physicians and public health departments for childhood immunization. AB - The purpose of the study was to examine factors influencing the use of primary care physicians and public health departments for childhood immunization for patients in rural and urban areas. A telephone survey employing probability sampling (random digit dialing) was conducted to obtain data from a sample of adults (> or = 18 years) living in Kentucky. Data are from 97 households with children under age 5 living in the home. The majority of the respondents (95%) reported that their children had received immunizations. The primary locations for receipt of immunizations were the health department (51%) and a primary care physician's office (37%). Sixty-five percent of those who used the health department for childhood immunizations reported that they did so for financial reasons. Individuals who received immunizations from the health department were more likely than those who received them at a primary care physician's office to have incomes at or below the poverty level and live in a rural area. The results of a logistic regression computed on use of the health department or primary care physician for immunizations indicated rural/urban residence as the only significant predictor, with urban residents 3.7 times more likely than rural residents to receive immunizations from a primary care physician. These results suggest that many families in rural areas have primary care physicians, but use the health department for their routine childhood immunizations. The results support previous data which indicate that delivery of childhood immunizations by primary care physicians is less available to rural than urban individuals. PMID- 8228582 TI - Apnea of infancy associated with Epstein-Barr virus infection. AB - The Epstein-Barr virus is one of the most common causes of communicable disease. The clinical manifestations of this virus may vary widely depending on a child's age when primary infection occurs. I describe in this report a child with apnea associated with Epstein-Barr infection. PMID- 8228583 TI - Fetal sex prediction: the heart of the matter. PMID- 8228584 TI - "Push the envelope". PMID- 8228585 TI - A lion in a den of Daniels. PMID- 8228586 TI - Results of the Maternal and Child Health Committee survey of family practitioners and obstetrics and gynecology specialists. PMID- 8228587 TI - KMA releases domestic/interpersonal violence educational packet. PMID- 8228588 TI - Paranasal aspergillosis--a spectrum of disease. PMID- 8228589 TI - Informed consent: using a structured interview changes patients' attitudes towards informed consent. AB - Patients want to know more about their condition and its proposed treatment. Gaining patients' confidence before treatment reduces the changes of their seeking legal redress for an unexpected outcome. As part of a prospective study of informed consent for surgery we have assessed the attitudes of patients towards informed consent when different types of consent interview are used. We found that most patients are happy to do as their doctor advises but think the informal consent interview is important because it gives them information; they also want to know about most, but not all, complications of the procedure. One quarter worried about the anaesthetic, about one eighth worried about 'not waking up' and similar proportions worried about complications and other things such as pain and nausea. Most patients think that the consent form is a legal document. In addition patients who had an informal interview felt obliged to sign the consent form and thought it had medicolegal implications. In contrast those who had a structured interview felt less obliged to sign the consent form and more involved in the decision to operate. PMID- 8228590 TI - Cortical evoked potential criteria in the objective assessment of auditory threshold: a comparison of noise induced hearing loss with Meniere's disease. AB - Amplitude of the N1 component of the cortical response was used to objectively determine threshold of hearing at 1 kHz and 4 kHz in a series of consecutively referred medicolegal cases with alleged occupational noise induced hearing loss and a control group of patients with Meniere's disease who were not seeking compensation for their hearing loss. The cortical response thresholds were compared with the subjective pure tone audiometric (PTA) thresholds at the same frequencies. The cortical and PTA thresholds were 'within 10 dB' for 84 and 92 per cent of the cases of noise induced hearing loss (NIHL) and Meniere's disease respectively, confirming the validity of CERA as a means of defining accurately the frequency specific thresholds and the audiometric configuration. Of the remaining 16 per cent of NIHL, 13 per cent exaggerated their PTA thresholds at 1 kHz and 10 per cent at 4 kHz whilst the error in cortical threshold estimation was beyond the 10 dB level for three and six per cent of cases at those frequencies respectively. The median exaggeration of threshold was 25 dB. For eight per cent of the Meniere's patients, thresholds exceeded 10 dB at both 1 and 4 kHz, four per cent of whom exaggerated their PTA thresholds and four per cent had a test error greater than 10 dB. A similar percentage (four per cent) of both groups revealed a cortical test error greater than 10 dB whereas three times as many cases of noise induced hearing loss (13 per cent) revealed exaggeration of their subjective audiometric thresholds compared with the Meniere's group (four per cent).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228591 TI - A comparison of speech audiometry and pure tone audiometry in patients with secretory otitis media. AB - Speech audiometry was performed on 15 children with secretory otitis media pre- and post-operatively and the findings compared with the pre- and post-operative pure tone audiograms. The results showed that in 30 per cent of cases pre operatively the speech audiogram suggested a significantly worse hearing ability than that suggested by the pure tone audiogram (PTA). Where a PTA gives a borderline threshold disproportionate parental concern should be taken seriously as the hearing ability may well be worse than the PTA suggests. PMID- 8228592 TI - Sensorineural hearing loss in sickle cell anaemia--a United Kingdom study. AB - Sensorineural hearing loss (SNHL) has been a well-documented complication of sickle cell disease in the literature from West Africa, West Indies, United States of America and the Middle East. We present a study of 52 patients with homozygous sickle cell disease and 36 control patients with haemoglobin genotype AA, matched for age and sex. Seven patients with sickle cell disease (13.5 per cent) were found to have sensorineural hearing loss i.e. > 20 dB at two or more frequencies, while all the patients in the control group had normal hearing (p < 0.05). Our study shows the incidence of SNHL in the UK to be similar to that reported in the USA and much lower than that found in malaria endemic areas of the tropics. We highlight the factors which we consider responsible for these differences and suggest that the crucial period in the development of SNHL in sickle cell disease may be intra-uterine or during the first few years of life. All sickle cell patients should be encouraged to have regular hearing assessment. PMID- 8228593 TI - Posterior crus stapedectomy. AB - The results of stapedectomy were compared between 162 otosclerotic ears operated on using the posterior crus technique and 182 otosclerotic ears undergoing Teflon piston stapedectomy. The large fenestra technique with fascia seal to the oval window was used in all cases. Mean follow-up period was 9.6 years. Neither short nor long-term hearing results showed any significant differences between the two surgical technique groups. Complications of surgery were more common in patients undergoing Teflon piston stapedectomy. On the other hand, a re-operation for recurring conductive deafness was performed significantly more often in patients undergoing posterior crus stapedectomy. PMID- 8228594 TI - The management of lateral sinus thrombosis. AB - The standard recommended treatment for lateral sinus thrombosis is intravenous antibiotics. For those that fail, some authors recommend anticoagulant therapy, others internal jugular vein ligation. Despite these recommendations, there is still a significant mortality in all reported large series and the main reason this is that the cause of the failure has not been found or investigated. Over a six-year-period, from January 1985 to December 1990, 36 patients were admitted with lateral sinus thrombosis. The initial treatment consisted of intravenous antibiotics and surgery. The surgery included cortical mastoidectomy for non cholesteatomatous ears and modified radical mastoidectomy for cholesteatomatous ears and drainage of the intracranial collection of pus in patients with subdural empyema and brain abscess. There were nine failures (25 per cent) with this treatment regimen. Blood cultures and pus swab of the ears were taken from these patients. The blood cultures were positive in four patients and the organisms isolated included beta-haemolytic Streptococcus sp. in two, Proteus mirabilis and Streptococcus pneumoniae in one each. From the ear swab mixed cultures were found in eight and a single organism in one. Gram-negative organisms were isolated in nine (100 per cent), Proteus mirabilis in eight (89 per cent) and Enterobacter sp. in two. Both these organisms were resistant to ampicillin and penicillin but were sensitive to amikacin. Amikacin was then added to the treatment regimen and a dramatic response with a fall in the temperature, and improvement in the patients' general condition was noted within 24 hours, in all patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228595 TI - Medical therapy of rhinosporidiosis with dapsone. AB - Clinical, histopathological and ultrastructural studies in three cases of rhinosporidiosis show complete remission of infection within one year of therapy with dapsone. Light and electron microscope studies confirmed arrest of the maturation of the spores and accentuated granulomatous response with fibrosis following dapsone therapy. PMID- 8228596 TI - Supraglottic laryngectomy: experience with 66 patients over 20 years. AB - We reviewed the records of 66 consecutive patients with previously untreated supraglottic laryngeal carcinoma who underwent a supraglottic laryngectomy in our department between 1970 and 1989. There were 23 T1, 30 T2 and 13 T3 cases. Eight patients had neck metastases and underwent neck dissection also. Twenty-four (36 per cent) patients received post-operative radiotherapy. Two-year and five-year survival rates were 88.9 per cent and 82.2 per cent respectively. When the survival rates of the patients with T1, T2 and T3 lesions were calculated separately and compared with each other the differences were not statistically significant. Recurrences, post-operative complications, time to progression and indications for supraglottic laryngectomy are discussed. PMID- 8228597 TI - Near-total laryngopharyngectomy with pectoralis major myocutaneous flap in advanced pyriform carcinoma. AB - Successful restoration of phonation, as well as swallowing in laryngopharyngeal surgery for patients with advanced pyriform carcinoma still remains a major challenge. In a five-year period, near-total laryngectomy operations were performed on 21 patients with laryngopharyngeal carcinoma. This report particularly emphasizes the value of near-total laryngopharyngectomy with a pectoralis major myocutaneous flap for four extensive hypopharyngeal carcinoma cases (three of which were of pyriform origin). All four patients regained intelligible shunt speech and smooth swallowing. The follow-up period ranged from 19 to 44 months. They have had no recurrence of disease since surgery. The authors suggest that an advanced pyriform carcinoma case, with a normal contralateral hemilarynx and pyriform sinus, is a candidate for a near-total excision of the laryngopharynx. PMID- 8228598 TI - Palpation compared with ultrasound in the assessment of malignant cervical lymph nodes. AB - This prospective study compares the results of pre-operative clinical examination and simple ultrasound examination of the neck in 18 patients with proven head and neck primary tumours and palpable cervical lymphadenopathy, who then underwent 21 radical neck dissections. Neck palpation and ultrasound examination were compared with histological examination. Ultrasound did not add significantly to the information obtainable by simple neck palpation in this group of patients. PMID- 8228599 TI - The tracheostomal washer: a simple aid for tracheo-oesophageal speakers with a large tracheostome. PMID- 8228600 TI - Laryngeal closure at the level of the false cord for the treatment of aspiration. AB - A new procedure for aspiration which closes the larynx at the level of the false cords is reported. This method is not harmful to the vocal folds and maintains arytenoid movement, thus preserving phonatory function. With the advance of the operative technique for aspiration and swallowing rehabilitation, patients who have recovered from aspiration can be helped. Our procedure can be recommended for such cases. PMID- 8228601 TI - External ear canal schwannoma: an unusual case report. AB - To the best of our knowledge, this is the first report of external ear canal schwannoma in the English literature. Several detailed clinical and pathological features were demonstrated. We suggest that if a tender, encapsulated mass is found in the external ear canal, the diagnosis of schwannoma should be taken into consideration. PMID- 8228602 TI - Sudden sensorineural hearing loss in haemoglobin SC disease. AB - Presented herein is a case report of sudden sensorineural hearing loss in the setting of haemoglobin SC disease. The relationship of the two is rare; the authors have found that this is only the second report in the literature. In this instance, partial exchange transfusions were performed in an attempt to decrease viscosity and improve blood flow. Thereafter, hearing stabilized and then slowly improved. The evidence for the beneficial role of these transfusions in this setting is, at best, circumstantial, but it is theoretically sound and worthy of further study. PMID- 8228603 TI - Nasal leishmaniasis. AB - Leishmaniasis is an uncommon condition in Western Europe, except around the Mediterranean coast. However, it may occasionally be seen in the United Kingdom, in patients who acquired the infection in foreign lands. An unusual case of localized cutaneous leishmaniasis recidiva affecting the nose after septal surgery is presented. PMID- 8228604 TI - Transnasal endoscopic surgery of sphenoid sinus aspergillosis. AB - Aspergillus sp. are the most common contaminants of the paranasal sinuses with the maxillary sinus as the site most frequently involved. Fungal sphenoid sinusitis can be a life-threatening situation, thus aggressive therapy is indicated. The treatment is primarily surgical. The transnasal endoscopic technique offers excellent visualization and an atraumatic approach to the sphenoid sinus. Surgical exteriorization and aeration of the involved sinus, without using antifungal chemotherapeutic agents is curative. A rare case of sphenoid sinus aspergillosis which was successfully treated by transnasal endoscopic sphenoidotomy is reported. PMID- 8228605 TI - Acute visual loss in association with sinusitis. AB - Acute visual loss may occur in association with sinusitis either as a complication of orbital cellulitis or, less frequently, as a part of the orbital apex syndrome. We describe two cases of temporary monocular visual loss caused by sinusitis. In one case the visual loss was due to orbital cellulitis; while in the other patient it was due to incompletely developed orbital apex syndrome. This later mode of presentation is called 'partial orbital apex syndrome' by some authors and 'posterior orbital cellulitis' by others. The relationship between sinusitis and blindness is discussed. PMID- 8228606 TI - Melanotic neuro-ectodermal tumour of infancy arising from the squamous and occipital bone. AB - The melanotic neuro-ectodermal tumour of infancy is an uncommon benign tumour of neural crest origin occurring in infants with a predilection for the anterior maxilla. This lesion shows a good response to conservative surgical excision with few recurrences but it should be followed up for long periods of time because of the rare malignant change possible in the tumour. PMID- 8228607 TI - Tuberculosis of the larynx in a lepromatous patient. AB - Within the black population of South Africa tuberculosis and leprosy are endemic. There is also a significant incidence of laryngeal carcinoma. A patient who presented in acute respiratory stridor to a rural hospital with limited resources is reported. The differential diagnosis and management is discussed in the light of the available literature. PMID- 8228608 TI - Malignant schwannoma of the parapharyngeal space in von Recklinghausen's disease: a case report and review of the literature. AB - Neurofibromatosis (NF) or von Recklinghausen's disease is frequently accompanied by malignant tumours, which can occur at any site in the body. These malignancies are mainly of soft tissue origin. Of all head and neck malignancies, the number of soft tissue sarcomas is limited and in combination with NF this type of tumour is a rare event. In this report we describe the clinical course of a young female patient with NF, who presented with a massive malignant schwannoma in the parapharyngeal space, and review the pertinent literature. PMID- 8228609 TI - Pigmented villonodular synovitis of the temporomandibular joint. AB - The first case of pigmented villonodular synovitis of the temporomandibular joint in a Chinese patient is reported. The clinicopathological features are described and the presentation as a parotid mass is emphasized. This rare tumour requires a high clinical suspicion for diagnosis. For removal, meticulous dissection of tumour and facial nerve is necessary. PMID- 8228610 TI - Cystic hygroma of the parotid gland. AB - Cystic hygroma of the parotid gland is an uncommon benign congenital neoplasm that presents as an asymptomatic soft fluctuant mass. Cervical ultrasonography, computerized tomography (CT) and magnetic resonance imaging (MRI) are used to assess the size and extent of the lesion and to assist in planning the surgical approach. Ultrasonography may demonstrate a multilocular cystic mass containing septa of variable thicknesses. Extension of the mass into deeper structures is more accurately assessed by CT or MRI. A CT scan may show thin-walled cystic masses filled with material of density close to that of water. The use of contrast medium can demonstrate the relationship of the lesion to the surrounding structures. Magnetic resonance imaging is superior to CT in defining the nature and extent of the lesion. PMID- 8228611 TI - Salivary neoplasms of the palate: a flow cytometric and clinicopathological analysis. AB - In order to test the clinical and prognostic significance of flow cytometrically assessed DNA content in minor salivary gland tumours we evaluated 75 neoplasms of the palate, 55 of which were carcinomas. Benign neoplasms were exclusively DNA diploid with low S-phase fractions while 22 per cent of malignant tumours manifested a DNA aneuploidy and 23.5 per cent high S-phase fractions (> 5 per cent). Significant statistical correlations between DNA content and tumour size, histological grade, lymph node metastasis and lethality were observed. Our findings suggest a potentially important role for flow-cytometry in the evaluation of these neoplasms. PMID- 8228612 TI - Basal cell adenocarcinoma of the parotid gland: a case report and review of the literature. AB - Basal cell adenocarcinoma is a rare salivary gland tumour. A DNA diploid parotid gland basal cell adenocarcinoma, with a low mitotic rate, in a 78-year-old female patient is described. Total removal of the affected salivary gland without sacrificing the facial nerve gave a good therapeutic result. PMID- 8228613 TI - Blessed be the tie that binds? Antitrust perils of physician investment and self referral. PMID- 8228614 TI - Neonatal HIV seroprevalence studies. A critique of national and international practices. PMID- 8228615 TI - Federal income taxation of nonprofit hospital joint ventures. New approach yields inconsistent results. PMID- 8228616 TI - NADPH-oxidase activity: the probable source of reactive oxygen intermediate generation in hemocytes of the gastropod Lymnaea stagnalis. AB - Macrophage-like defense cells (hemocytes) of the pond snail Lymnaea stagnalis generate reactive oxygen intermediates (ROIs) upon contact with non-self, following kinetics similar to those of ROI production by mammalian leukocytes during respiratory burst. In this study, several inhibitors of NADPH-oxidase, the key enzyme of the respiratory burst in mammalian phagocytes, were tested for their effect on oxidative activities [as demonstrated by nitroblue tetrazolium (NBT) reduction and luminol-dependent chemiluminescence (LDCL)] of phagocytosing snail hemocytes. In the presence of diphenylene iodonium, zymosan-stimulated hemocytes of L. stagnalis failed to reduce NBT and showed a markedly reduced LDCL response. Also, compounds that prevent assembly of functional NADPH-oxidase complexes in activated mammalian cells were effective; preincubation of hemocytes with 1,4-naphthoquinone inhibited the LDCL response and NBT reduction upon phagocytic stimulation. Furthermore, coincubation but not preincubation with five different catechol-like phenols inhibited oxidative activities of zymosan stimulated hemocytes. These results imply similarities in composition and regulation of the ROI-generating mechanisms of both mammalian and snail defense cells. It is postulated that in L. stagnalis hemocytes, (1) NADPH-oxidase activity is responsible for ROI production, (2) an active NADPH-oxidase enzyme complex has to be assembled from putative cytosolic and membrane-associated components, and (3) continuous replacement of active NADPH-oxidase enzyme complexes is necessary to sustain respiratory burst-like oxidative activities during interactions with non-self. PMID- 8228617 TI - Lipopolysaccharide and a phorbol ester stimulate secretion of tumor necrosis factor-alpha from alveolar macrophages through action on overlapping subsets of cells. AB - A cell-by-cell analysis of the secretory ability of stimulated, individual alveolar macrophages (AMs) was performed through use of a tumor necrosis factor alpha (TNF-alpha) reverse hemolytic plaque assay. Two functional end points were measured: the percentage of AMs that were TNF-alpha secretors and the cumulative amount of TNF-alpha secreted by AMs (average plaque area, microns 2). Lipopolysaccharide (LPS; 100 micrograms/ml) increased cumulative TNF-alpha release at both 7 and 20 h of incubation. On the other hand, a phorbol ester (phorbol myristate acetate, PMA) stimulated TNF-alpha release at 20 h of incubation but not at 7 h. Under nonstimulated culture conditions, 5-10% of all AMs released detectable TNF-alpha PMA (but not LPS) induced a significant increase in the fraction of AMs capable of releasing TNF-alpha (15.1 +/- 1.1% vs. 9.0 +/- 1.6%, PMA vs. control, P < .05). Differences in the time course of secreted TNF-alpha, together with the recruiting effect of PMA, suggest that LPS and PMA target TNF-alpha-secretory subpopulations of AMs that differ in number and secretory characteristics. PMID- 8228618 TI - Immunoglobulin G-mediated phagocytosis activates a calcium-independent, phosphatidylethanolamine-specific phospholipase. AB - Inhibition of arachidonate release down-regulates immunoglobulin G-mediated phagocytosis. This arachidonate requirement is selective for IgG-opsonized targets, suggesting that arachidonate may act as a second messenger for Fc gamma receptor-mediated phagocytosis. Here we report the characterization of a phospholipase, activated during phagocytosis, that releases arachidonate from phosphatidylethanolamine in the absence of intracellular calcium ([Ca]i < or = 2 nM). In vitro, a phospholipase with these characteristics was detected in soluble and particulate fractions of human monocyte homogenates. (E)-6 (Bromomethylene)tetrahydro-3-(1-naphthalenyl)2H-pyran-2-one, a drug that selectively inhibits Ca-independent phospholipase A2s, is shown to inhibit IgG mediated phagocytosis and its associated arachidonate release in intact monocytes as well as the in vitro enzyme activity. These findings provide a link between the whole-cell and in vitro data and present the initial characterization of a receptor-activated, calcium-independent phospholipase from human monocytes. PMID- 8228619 TI - Silica induces apoptosis in macrophages and the release of interleukin-1 alpha and interleukin-1 beta. AB - Resident adherent peritoneal cells selectively released high amounts of interleukin-1 (IL-1) activity when treated with silica. The use of anti-IL-1 antisera showed that both IL-1 alpha and IL-1 beta were present in supernatants of silica-treated macrophages. In contrast, intracellular IL-1 activity was totally neutralized by anti-IL-1 alpha antibodies and was easily converted into the mature IL-1 alpha form by autolysis in cytoplasmic extracts. Anion exchange chromatography clearly separated the two IL-1 species present in supernatants of silica-stimulated macrophages. Natural IL-1 beta was further characterized by chromatofocalization; it had an apparent isoelectric point, pI, in the range 8.3 8.6. In agreement with previous findings showing that IL-1 beta was released only by apoptotic cells, we have found that silica-treated macrophages underwent apoptosis. This was demonstrated by the characteristic laddering electrophoretic pattern of DNA extracted from silica-treated cells and by the morphology of macrophage nuclei stained with the DNA-specific dye DAPI. In addition, quantification of apoptotic cells was performed by a flow cytometric analysis based on the reduction of cellular DNA content exhibited by apoptotic cells. Treatment of macrophages with silica, therefore, results in an active process that promotes the processing and liberation of IL-1 beta. PMID- 8228620 TI - Transforming growth factor beta 1 regulation of macrophage activation depends on the triggering stimulus. AB - We have examined the effects of transforming growth factor beta 1 (TGF-beta 1) on the regulation of murine bone marrow-derived macrophage function. TGF-beta, added simultaneously with or up to 4 h before interferon-gamma (IFN-gamma) plus lipopolysaccharide (LPS), inhibited macrophage leishmanicidal activity, nitrite (NO2-) production, and secretion of prostaglandin E2. In contrast, no effect of TGF-beta could be demonstrated on macrophages stimulated with IFN-gamma plus tumor necrosis factor-alpha (TNF-alpha) under the same conditions. These results suggested that TGF-beta inhibited LPS-induced triggering of macrophage activation, which was confirmed by studies with IFN-gamma-primed cells. Interestingly, when macrophages were pretreated with TGF-beta for 24 h, NO2- production in response to IFN-gamma plus TNF-alpha was also inhibited. Although control and IFN-gamma/LPS-stimulated macrophages were found to secrete latent TGF beta, only the IFN-gamma/LPS cultures produced biologically active TGF-beta. Significantly, active TGF-beta was present at concentrations shown earlier to inhibit macrophage function. PMID- 8228621 TI - Preincubation with anti-CD4 influences activation of human T cells by subsequent co-cross-linking of CD4 with CD3. AB - Under physiological conditions, T cell activation by major histocompatibility complex (MHC)-antigen complexes requires engagement of both the T cell receptor (TcR) and the CD4 (or CD8) accessory molecules. It has been shown, however, that ligation of CD4 and CD8 can also inhibit T cell activation in an MHC-independent way. Therefore, the role of CD4 in T cell activation and the mechanism of the suppression of T cell functions by anti-CD4 are as yet unclear. We activated T cells by CD4/CD3 co-cross-linking and studied the effect of preincubation with anti-CD4 on this activation. We show here that anti-CD4 effects T cell activation in a complex, time-dependent manner. Whereas short preincubations with anti-CD4 usually enhanced T cell proliferation in response to subsequent co-cross-linking of CD3 with CD4, longer preincubations led to its decrease. The observed suppression of proliferation after a long preincubation with anti-CD4 was apparently due to impairment of TcR signaling, as assessed by measurement of Ca2+ mobilization and tyrosine phosphorylation in T cells. These results add a temporal element to the previously observed synergism between the TcR and CD4 in T cell activation. PMID- 8228622 TI - Particle-induced in vivo priming of alveolar macrophages for enhanced oxidative responses: a novel system of cellular immune augmentation. AB - A novel system for priming adult rabbit alveolar macrophages (AMs) in vivo for markedly enhanced oxidative responses is described. When adult rabbits were injected intravenously (i.v.) with 1- to 5-microns particles such as zymosan, latex particles, or heat-killed bacille Calmette-Guerin, AMs were primed in 1-3 days for greatly enhanced phorbol myristate acetate (PMA)- or opsonized zymosan (Op-zym)-elicited chemiluminescent (CL) responses. Intratracheal (i.t.) injection of zymosan particles also primed AMs for enhanced PMA- or Op-zym-elicited CL responses. AMs obtained from particle-injected rabbits showed up to 100-fold higher levels of PMA-elicited CL responses than AMs from normal rabbits. In contrast, Op-zym failed to prime normal AMs in vitro for enhanced CL responses. Whereas AMs could not be primed in vivo with an i.v. injection of particles of approximately 24 microns diameter. AMs could be primed if the particles were administered by the i.t. route. The priming appears to be independent of particle types. The priming effect was of short duration and declined after 5 to 7 days. The possibility that this system represents the primitive cellular immune response found in invertebrates is discussed. The potential use of this system as a means of immune augmentation prompts further investigation. PMID- 8228623 TI - Lipopolysaccharide structure-function relationship in activation versus reprogramming of mouse peritoneal macrophages. AB - Lipopolysaccharide (LPS) is one of the most potent stimuli for macrophages. The activities of LPS have been attributed to the lipid A region of the molecule. We have previously shown that pretreatment of macrophages with very low doses of LPS can selectively "reprogram" these cells to respond differentially to subsequent activation, as assessed by tumor necrosis factor-alpha and nitric oxide (NO) production. Here we demonstrate that the relative capacity of various LPS preparations for induction of down-regulation of subsequent LPS-activated NO production correlates well with their relative potency for initiation of NO formation. Although LPS-dependent activation can be regulated by pertussis toxin (PT)-sensitive factor, the LPS pretreatment-induced reprogramming is shown here to be refractory to regulation by PT. These results suggest that, although the structural components of LPS dictating the relative activities of the molecule for activation versus reprogramming are similar, there may exist different pathways in initiation of LPS-induced activation versus reprogramming. PMID- 8228624 TI - Differential effects of interferon-gamma and glucocorticoids on Fc gamma R gene expression in murine macrophages. AB - Interferon-gamma (IFN-gamma) was shown previously to increase Fc gamma receptor (Fc gamma R)-mediated binding and phagocytosis of immunoglobulin G-opsonized erythrocytes in mouse peritoneal exudate macrophages. Glucocorticoids potentiated this effect. We have extended these observations to an investigation of the effects of IFN-gamma and glucocorticoids on steady-state mRNA levels of the three Fc gamma R genes expressed on murine macrophages: Fc gamma RI, Fc gamma RII, and Fc gamma RIII alpha. Fc gamma RI mRNA was present at a barely detectable level in unstimulated cells but was induced 5- to 10-fold by IFN-gamma. Fc gamma RIII alpha mRNA was expressed constitutively and was induced significantly but very modestly (1.5-fold) by IFN-gamma. Fc gamma RII mRNA also exhibited high constitutive expression, but it was not altered by IFN-gamma treatment. Dexamethasone (DEX) significantly increased the expression of IFN-gamma-induced Fc gamma RI mRNA but had no effect on the expression of Fc gamma RII or Fc gamma RIII alpha transcripts in untreated or IFN-gamma-treated cells. The effect of DEX on Fc gamma RI mRNA was seen after about 10 h of simultaneous treatment with IFN gamma, was dose-dependent, and was glucocorticoid specific. DEX did not modulate the rate of decay of the Fc gamma RI mRNA, suggesting that the up-regulatory effect of DEX may occur, in part, at the level of transcription. In the same culture system, the steady-state level of IFN-gamma-induced Ia mRNA was concurrently inhibited by DEX. The up-regulation of the IFN-gamma-induced high affinity Fc gamma RI mRNA, the failure to modulate the expression of the two low affinity Fc gamma R mRNA species, and the down-regulation of IFN-gamma-induced Ia mRNA by DEX in the same cell population illustrate the diverse, gene-specific influence of glucocorticoids on the expression of IFN-gamma-inducible genes. PMID- 8228625 TI - Cytotoxic cell proteinase gene expression and cytolytic activity by anti-CD3 activated cytotoxic T lymphocytes is sensitive to cyclosporin A but is not dependent on interleukin-2 synthesis. AB - We have examined the role of interleukin (IL) 2 in the expression of cytotoxic cell proteinases (CCP) 1 and 2, as well as in the induction of major histocompatibility complex (MHC)-unrestricted cytotoxic activity in murine T cell cultures following stimulation with anti-CD3 monoclonal antibody. A dramatic reduction in CCP-1 and CCP-2 gene expression and near absence of cytolytic activity was shown to occur in these cultures when the expression of IL-2 was inhibited by 10(-6) M cyclosporin A (CsA). The inhibitory effect of CsA could not be eliminated by the addition to culture of recombinant IL-2 at concentrations typically present in anti-CD3-stimulated T cell culture supernatants. Furthermore, when endogenous IL-2 (45-60 U/ml) present in anti-CD3-stimulated T cell cultures was neutralized with anti-mouse IL-2 antibody there was no effect on CCP-1 and CCP-2 mRNA expression and only a slight decrease in cytolytic activity. The expression of CCP-1 and CCP-2 gene products and the induction of MHC-unrestricted cytotoxic activity in anti-CD3-stimulated T cell cultures therefore occur independently of IL-2 synthesis but are regulated by a CsA sensitive mechanism. PMID- 8228626 TI - Random depletion of T cells that bear specific T cell receptor V beta sequences in AIDS patients. AB - A cross-sectional PCR analysis of the TCR V beta repertoires in HIV-1 seronegative controls and HIV-1 infected individuals with either clinically or immunologically defined AIDS [1] was performed to examine the proposed superantigen model for HIV-1 pathogenesis. In contrast to previous reports, we find neither uniform specific losses nor uniform clonal expansions of particular TCR V beta gene families in subjects with AIDS. Instead our study, which was designed specifically to qualitatively determine the presence or absence of TCR V beta families in both subject populations, indicates an overall diminution in the expression of TCR V beta gene families in HIV-1 infected individuals with AIDS compared with controls. This is commensurate with the decrease in CD4 T cells in the AIDS population. Our data are therefore not directly suggestive of a common superantigen model of HIV-1 induced T cell clonal depletion or anergy, but instead emphasize a broad decrease in signals throughout the TCR V beta repertoire in AIDS versus control groups. This random depletion in the TCR V beta repertoire is most likely caused by aspects of HIV-1 pathogenesis other than virus-encoded superantigens. PMID- 8228627 TI - Receptor-receptor interactions of complement receptor type 3 in neutrophil membranes. AB - The leukocyte integrin CR3 (CD11b/CD18) is known to participate in a variety of cell functions. Recent studies have indicated that CR3 may communicate with other plasma membrane receptors to carry out several cell functions. In this review we discuss these potential receptor-receptor interactions of CR3 and present a unifying model of CR3's diverse functions. PMID- 8228628 TI - Interplay between superantigens and the immune system. AB - Superantigens interact with the immune system by binding to major histocompatibility complex (MHC) class II proteins and activating T cells through the variable region of the T cell receptor beta-chain. Through this means they can cause massive proliferation and then death of a large proportion of T cells. Superantigens are produced by bacteria, mycoplasmas, retroviruses, and probably by other organisms. In some cases, the superantigen is crucial to the organism's life cycle. Mouse mammary tumor virus disseminates by activating T cells which stimulate the proliferation of B cells harboring the virus. In other cases, the superantigen may be responsible for the pathogenesis of the infection, such as in the case of Toxic Shock Syndrome. In this article, we review information on the diseases in which superantigens are involved, and the mechanisms by which the superantigens interact with T cell receptor and class II molecules. PMID- 8228629 TI - Soluble Fc gamma receptors. AB - Soluble Fc gamma receptors have been identified in biological fluids of mice and humans. They are produced either by alternative splicing of the exon encoding the transmembrane region of the receptor (Fc gamma RII) or by proteolytic cleavage at the cell membrane (Fc gamma RII and Fc gamma RIII). They inhibit B cell proliferation and immunoglobulin production. Their concentrations in plasma seem to be modified during the development of certain diseases, as for instance in multiple myeloma, where plasma concentrations of soluble Fc gamma RIII are correlated with the stage of the disease. PMID- 8228630 TI - Effect of phospholipids and bile acids on cholesterol nucleation time and vesicular/micellar cholesterol in gallbladder bile of patients with cholesterol stones. AB - Supersaturation and rapid nucleation of cholesterol in bile are of key importance in the pathogenesis of cholesterol gallstones. While the effects of bile acids and phospholipids on cholesterol saturation of bile have been extensively studied, their influence on the cholesterol nucleation time has not been compared. We, therefore, investigated whether increases of bile acid or phospholipid concentrations in bile by in vitro supplementation affect the cholesterol nucleation time. Bile samples were obtained at surgery from patients with cholesterol gallstones. Prior to the nucleation assay the bile samples were divided into 0.5-ml aliquots and supplemented with 1.25, 2.5, 5.0, and 10.0 mumol/ml of different phosphatidylcholines (PC-dimyristoyl, PC-dipalmitoyl, PC distearoyl, and extracted biliary PCs) or with 5.0, 10.0, and 20.0 mumol/ml of bile acids (glycine or taurine conjugates of cholic acid, deoxycholic acid, or chenodeoxycholic acid). The increase of phosphatidylcholine or bile acid concentration decreased the mean cholesterol saturation index to a similar extent (PC: 0.1-0.3; BA: 0.1-0.2). Supplementations of bile with increasing amounts of synthetic or biliary PCs caused a marked prolongation of the nucleation time in bile from 1.5 +/- 0.2 up to > or = 21 days or 2.5 +/- 0.7 up to > or = 21 days. Concurrently, biliary cholesterol was shifted from vesicles to mixed micelles and the cholesterol/phospholipid ratio of the remaining vesicles was progressively lowered. In contrast, the addition of bile acids to gallbladder bile did not affect the cholesterol nucleation time (2.2 +/- 0.3 days), the percentage of vesicular cholesterol, or the cholesterol/phospholipid ratio of vesicles and micelles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228631 TI - Lipoprotein structure in male subjects during in vivo lipolysis: effect of an anti-lipolytic treatment with acipimox. AB - Plasma free fatty acid (FFA) levels were raised in healthy volunteers by the administration of a fatty meal and epinephrine infusion (0.15 mg/kg per min), to test the hypothesis that enhanced lipolysis might lead to changes in lipoprotein distribution and to the formation of lipoprotein complexes, also impairing the interconversion of high density lipoproteins (HDL). The study was carried out in double-blind conditions in volunteers pre-treated with either placebo or with acipimox, a nicotinic acid analogue with a long-lasting activity. Lipolysis was effectively induced; the treatment with acipimox prevented the rise of free fatty acids (FFA), and it also blunted the triglyceride (TG) increase occurring during the test. Whereas the mean low density lipoprotein (LDL) particle size did not change, the HDL particle distribution showed a progressive shift to smaller particles, both after placebo and after acipimox, the changes in size being maximal 3-7 h after the meal. Evaluation of HDL interconversion in plasma samples incubated at 37 degrees C for 6 h showed the expected accumulation of HDL2a particles, with a parallel decrease of HDL3a; however, this conversion was not affected by the presence of elevated FFA levels and no difference was noted in subjects taking either placebo or acipimox. These clinical data fail to confirm the hypothesis that enhanced lipolysis may lead to dramatic changes in plasma lipoprotein distribution and/or in aggregation or fusion of lipoprotein particles, as reported from in vitro experiments. This study, however, successfully achieved a useful model of exaggerated lipolysis and confirmed the important activity of a low dose nicotinic acid analogue in inhibiting lipolysis. PMID- 8228632 TI - Synthesis of monohydroxylated fatty acids from linoleic acid by rat aortic smooth muscle cells and tissues: influence on prostacyclin production. AB - We have investigated whether cellular metabolism of linoleic acid (18:2) can influence prostacyclin (PGI2) production by cultured rat aortic smooth muscle cells (SMC) and tissues. Incubation of rat SMC homogenates with [1-14C]18:2 results in the enzymatic synthesis of [14C]13-HODE (hydroxyoctadecadienoic acid) and to a lesser extent [14C]9-HODE as defined by gas-liquid chromatography-mass spectrometry (GLC-MS). The observed changes, in percent enzymatically synthesized 13-HODE in the presence of indomethacin, aspirin, metyrapone, 15-HPETE (hydroperoxyeicosatetraenoic acid), and NDGA, suggest that it is formed from the PGH (prostaglandin endoperoxide) synthase pathway. Incubation of intact adherent SMC with [14C]linoleic acid demonstrates that the monohydroxylated compounds are predominantly esterified within the membrane phospholipids and not released into the incubation medium. The simultaneous incubation or a short-term preincubation of 18:2 and arachidonic acid (20:4) do not modify the enzymatic profile of 20:4 transformation. By contrast, long-term preincubation of cells with 18:2 or 13 HODE stimulates the transformation of exogenously added [14C]20:4 to [14C]6-keto PGF1 alpha. However, exogenous 13-HODE does not enhance [14C]6-keto PGF1 alpha recovery from [14C]20:4 prelabeled SMCs. Our results demonstrate that 18:2 is a substrate for PGH-synthase in rat aortic SMC and tissues. The 13-HODE formed is essentially esterified in cell phospholipids and remains without any significant effects on the release of [14C]6-keto PGF1 alpha from [14C]20:4 prelabeled SMC. PMID- 8228633 TI - Radioiodination of low density lipoprotein initiates lipid peroxidation: protection by use of antioxidants. AB - It is now apparent that low density lipoprotein (LDL) is very susceptible to lipid peroxidation and that the resulting oxidized LDL has altered biological properties. Radiation, particularly of longer duration and lower intensities, initiates lipid peroxidation, yet radioiodination with 125I and 131I is a frequently used method to label LDL for biological studies. To test the possibility that this procedure alters the biological properties of LDL, native LDL was radioiodinated with 125I/131I using ICl to average specific activities of approximately 300 and approximately 100 cpm/ng protein, respectively. Lipid peroxidation was monitored by TBARS and conjugated diene formation. Biological properties were monitored by fibroblast and macrophage uptake of LDL as well as by rate of plasma clearance (FCR) in guinea pigs. 131I-labeled LDL showed enhanced indices of lipid peroxidation compared to 125I-labeled LDL and both were greater than native LDL. The FCR of 131I-labeled LDL was greater than that of 125I-labeled LDL (by 20-40%) and both increased progressively (by > 250%) when measured at 2, 6, and 13 days after iodination. The radioiodinated LDL samples were also more susceptible to pro-oxidant conditions. Thus, after exposure to Cu2+, 131I-labeled LDL showed greatly enhanced lipid peroxidation, decreased uptake by fibroblasts, increased uptake by macrophages and greatly accelerated FCR in guinea pigs. Exposure of LDL to 131I-labeled albumin produced similar changes. Protecting LDL with antioxidants such as BHT and ascorbate immediately after radioiodination generally ameliorated the adverse effects. PMID- 8228634 TI - Fatty acid oxygenase activity of human hair roots. AB - The extent to which fatty acid oxygenases are activated in the normal epidermis is not known. Characterization of the regio- and stereospecificity of the monohydroxylated derivatives of arachidonic and linoleic acid produced by human hair roots is needed to define the enzymatic origin of these compounds and to define a possible role for fatty acid oxygenases in growth, differentiation, and pathology of human hair. Hair roots epilated from normal human volunteers were incubated with radiolabeled arachidonic acid or linoleic acid and the monohydroxylated derivatives produced in vitro were characterized. Incubation of hair roots with 14C]arachidonic acid resulted in the production of 15(S) [14C]hydroxyeicosatetraenoic acid and 12(S,R)-[14C]hydroxyeicosatetraenoic acid (mean S/R ratio, 2.5). 13(S)-[14C]hydroxyoctadecadienoic acid was the principal product of incubations with [14C]linoleic acid. No radiolabeled products were derived from incubations with heat-denatured hair roots. The fatty acid oxygenase activity of anagen hair roots was inhibited by nordihydroguaiaretic acid and was greatest in the hair root bulb. The strict S-stereospecificity and the regiospecificity of the n-6 oxygenase are strong evidence for the presence of a 15-lipoxygenase in human hair roots, similar to that identified in cultured human keratinocytes. The stereospecificity of the 12-HETE produced by human hair roots is not compatible with the sole action of 12-lipoxygenase. PMID- 8228635 TI - Differential mobilization of white adipose tissue fatty acids according to chain length, unsaturation, and positional isomerism. AB - The present study aims at determining whether, and how, the molecular structure of fatty acids influences their mobilization from fat cells. The in vitro mobilization of 52 fatty acids ranging in chain length from 12 to 24 carbon atoms, in unsaturation from 0 to 6 double bonds, and including 23 pairs of positional isomers was examined. Fat cells were isolated from adipose tissue of rats fed a fish-oil diet and treated with norepinephrine to stimulate lipolysis. Fatty acid composition of free fatty acids (FFA) released from these cells was compared to that of the triacylglycerols (TAG) from which they originated. The percentage weight of most fatty acids, and especially that of very long-chain fatty acids, was significantly different between FFA and TAG. The percentage of 20:5n-3 (eicosapentaenoic acid) and of 20:4n-6 (arachidonic acid) was 2.7 and 1.7 times higher in FFA than in TAG, respectively, whereas that of 20:1(n-11, 9 or 7), 22:1(n-11, 9 or 7) and 24:1n-9 was 1.7-2, 2.4, and 3.5 times lower, respectively. The relative mobilization (% in FFA/% in TAG) of the least readily mobilized fatty acid (24:1n-9) was 15-fold lower than that of the most readily mobilized (18:5n-3). For a given chain length, the relative mobilization increased exponentially with unsaturation, e.g., increasing from 0.45 to 2.7 in C 20 fatty acids when the number of double bonds increased from 0 to 5. Amongst the fatty acids with 18 to 22 carbon atoms, the shorter the chain was, the more steeply relative mobilization increased with unsaturation. On the other hand, for a given unsaturation the relative mobilization decreased with increasing chain length, e.g., decreasing from 1.15 to 0.3 in monoenes when the chain length increased from C 14 to C 24. A two-carbon-atom shortening of the chain length was on average equivalent to inserting one double bond for increasing the relative mobilization, i.e., by about 40%. The relative mobilization was also affected by the position of the double bond(s); increasing on average by 10% when there was a two-carbon-atom displacement towards the methyl end of the chain. These results demonstrate that under conditions of stimulated lipolysis individual fatty acids are more readily mobilized from fat cells when they are short and unsaturated, and when their double bonds are closer to the methyl end of the chain.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8228636 TI - Serum cholesterol, cholesterol precursors, and plant sterols in hypercholesterolemic subjects with different apoE phenotypes during dietary sitostanol ester treatment. AB - A randomized double-blind study was made in 67 modestly hypercholesterolemic subjects by replacing 50 g of daily dietary fat by the same amount of a rapeseed oil preparation without and with fat-soluble sitostanol esters. The diet became relatively rich in dietary fat (37%) especially in subjects with a low basal calorie intake. The esters were prepared by transesterification of sitostanol with rapeseed oil fatty acids. The effects of sitostanol esters were studied on serum cholesterol and cholesterol synthesis (measuring cholesterol precursors in serum) and absorption (measuring serum plant sterols). The results were related to different apoE phenotypes. A 6-week regimen of about 3.4 g/day of sitostanol lowered total and low density lipoprotein (LDL) cholesterol levels by 7.5% and 10%, respectively, over that due to rapeseed oil alone. High density lipoprotein (HDL) cholesterol and triglyceride concentrations were unchanged. Thus, the HDL/LDL cholesterol ratio was significantly increased. The decrease in LDL cholesterol level was more consistent in subjects with the epsilon 4 allele than in those with homozygous epsilon 3 alleles. Sitostanol markedly decreased serum campesterol (-46%) and sitosterol (-30%), especially in subjects with the epsilon 4 alleles known to have high cholesterol absorption . The decreases of LDL cholesterol and plant sterols were interrelated, suggesting that reduced cholesterol absorption contributed to the lowering of LDL cholesterol. Serum sitostanol was unchanged, while the serum cholesterol precursors, delta 8 cholestenol, desmosterol, and lathosterol, were compensatorily increased by 10% (P < 0.05), most consistently in the subjects with epsilon 4 alleles, indicating an increase in cholesterol synthesis. The study demonstrates that sitostanol esters dissolved in dietary fat can be recommended for treatment of modest primary hypercholesterolemia and are apparently practical and suitable for cholesterol lowering in a general population. PMID- 8228637 TI - Intestinal vitamin A metabolism: coordinate distribution of enzymes and CRBP(II). AB - In the mucosal layer of the small intestine, we found nearly identical gradients of CRBP(II), retinal reductase, and LRAT levels down the duodenal-ileal axis, suggesting coordinate regulation of these three proteins. In all cases the level of binding protein or enzyme activity was greatest in the proximal intestine and then decreased sharply in the distal half. This pattern fits with the known capacity of the intestine to absorb vitamin A. In addition, the retinal reductase activity was found predominantly in the intestinal mucosa, while LRAT activity was found in both the intestinal mucosa and muscle. An even distribution of LRAT activity along the longitudinal axis of the intestinal muscle was consistent with an even distribution of CRBP in that tissue. In conjunction with LRAT activity and CRBP, we found endogenous retinyl ester stores in the intestinal muscle layer. The patterns of retinyl ester produced by LRAT in vitro and found in vivo were similar, with retinyl palmitate predominating and a high percentage comprised of retinyl stearate. We also observed a bile salt-independent retinyl ester hydrolase activity in intestinal muscle whose distribution paralleled the retinyl ester stores and LRAT levels. This hydrolase appears to be distinct from retinyl ester hydrolases described from other organs as its activity was insensitive to retinyl ester chain length, the presence of bile salts, or the addition of apo-CRBP. This activity was inhibited by diethyl-p-nitrophenyl phosphate (IC50 100 microM) and diethylpyrocarbonate (IC50 10 microM), demonstrating a requirement for active serine and histidine residues. In addition, we describe an activity present in some intestinal microsomal preparations that can perturb determinations of reductase and LRAT activity and must be avoided. PMID- 8228638 TI - Lipoprotein lipase in human plasma is mainly inactive and associated with cholesterol-rich lipoproteins. AB - This study was designed to further ascertain the presence in plasma of lipoprotein lipase (LPL) bound to circulating lipoproteins. Lipoprotein lipase mass and activity values in preheparin plasma from 20 volunteers were 69.8 +/- 6.6 ng.ml-1 and 1.54 +/- 0.15 mU.ml-1, respectively, and no significant correlation between mass and activity was observed. Fifteen min after heparin injection, LPL mass had increased to 536 +/- 60 ng.ml-1 and LPL activity to 261 +/- 34 mU.ml-1 and a highly significant correlation between the increments in mass and activity was observed. The released material had a specific activity of 0.57 +/- 0.03 mU.ng-1. The LPL mass in preheparin plasma eluted early from heparin-Sepharose, in the position expected for inactive LPL monomers. Western blot analysis showed that the eluted material had the size expected for the LPL subunit (55 kDa). The increment of mass and activity after heparin eluted later from heparin-Sepharose, in the position expected for active LPL dimers. It is concluded that preheparin plasma contains substantial amounts of inactive LPL protein, and that heparin releases mainly active LPL into circulation. On gel filtration LPL activity and mass in postheparin plasma eluted mainly in the positions of LDL and HDL. Electron microscopy of immunostained fractions showed reaction for LPL and apolipoprotein B, or apolipoprotein A-I, on the same particles. LPL mass in preheparin plasma eluted in a similar pattern, associated with LDL and HDL. In postprandial plasma substantial amounts of LPL protein eluted with the triglyceride-rich lipoproteins. When 125I-labeled bovine LPL was added to plasma or to ultracentrifugally isolated lipoproteins and then analyzed by gradient gel electrophoresis, the labeled lipase moved with the lipoproteins. The presence of substantial amounts of inactive LPL protein associated with lipoproteins in plasma may have important implications for the metabolism of the particles in view of recent reports on avid binding of LPL-lipoprotein complexes to cell surfaces and receptors. PMID- 8228639 TI - Characterization of a hamster melanoma-associated ganglioside antigen as 7-O acetylated disialoganglioside GD3. AB - We previously reported a hamster animal model of melanoma in which the tumor tissue expresses gangliosides GM3, GD3, and O-acetyl GD3. This ganglioside pattern is similar to that in human melanomas (Ren, S., A. Slominski, and R. K. Yu. 1989 Cancer Res. 49: 7051). In this study, we isolated and purified these gangliosides using chloroform-methanol extraction, Folch partition, chromatographies on DEAE-Sephadex A-25, and Iatrobeads columns. The yields of gangliosides GM3, GD3, and O-acetyl GD3 were 44.1 mg, 19.6 mg, and 9 mg per 100 g of Ma melanotic melanoma tissues, respectively. The structures of these gangliosides were characterized by periodate oxidation, gas chromatographic (GC) analysis, fast-atom bombardment-mass spectrometry (FAB-MS), and nuclear magnetic resonance (NMR) studies. The structure of hamster melanoma O-acetyl GD3 is different from the 9-O-acetyl GD3 previously reported in human melanoma. The major fatty acids of this ganglioside are C16:0, C18:0, C20:0, C22:0, and C24:0 and the long-chain base is C18-sphingosine. PMID- 8228640 TI - Lipid accumulation in isolated perfused rat hearts has no apparent effect on mechanical function or energy metabolism as measured by 31P NMR. AB - Male Sprague-Dawley rats were fed diets that contained 20% by weight soybean oil or rapeseed oil (21% and 43% erucic acid) for 7 days. The rapeseed oil diets increased the cardiac triacylglycerol content 5-fold and 25-fold, respectively, above control values. Hearts were removed from the animals and perfused with modified Krebs-Henseleit buffer at 37 degrees C. The calculated rate-pressure product was used as a measure of contractile function. 31P NMR spectra were acquired throughout a protocol that consisted of 12 min control perfusion, followed by 12 min perfusion with 20 microM isoproterenol, 12 min washout, 12 min total global ischemia, and 28 min reperfusion. The steady state levels of creatine phosphate, ATP, intracellular pH, contractile function, and the free energy of ATP hydrolysis (delta GATP) were determined for all three groups of hearts. Isoproterenol more than doubled the rate-pressure product of the hearts on all diets and decreased the concentrations of creatine phosphate and ATP with a concomitant rise in Pi. After global ischemia, creatine phosphate levels recovered fully, ATP levels remained low, and most hearts developed ventricular fibrillation. Changes in intracellular pH were the same for all groups: pH was 7.1 throughout the equilibration and isoproterenol perfusion period, decreased to pH approximately 6.4 during ischemia, and returned to 7.0 during reperfusion. The results indicate that the fat accumulation that occurs in the hearts of rats fed diets rich in high erucic acid rapeseed oil does not interfere with the cardiac high energy phosphate metabolism or contractile function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228641 TI - A systematic analytical chemistry/cell assay approach to isolate activators of orphan nuclear receptors from biological extracts: characterization of peroxisome proliferator-activated receptor activators in plasma. AB - Using a novel combination of analytical chemical and molecular biological techniques, lipophilic components of human plasma separated according to their physico-chemical properties were screened for their ability to activate the rat peroxisome proliferator-activated receptor (rPPAR). Activation of an rPPAR/glucocorticoid receptor chimera stably expressed in CHO cells by fractions in the initial screening guided further subfractionation. Characterization of an active subfraction by gas chromatography alone and in combination with mass spectrometry (GC-MS), indicated the presence of free fatty acids. Individual active components in this mixture were isolated by a final fractionation using high performance liquid chromatography (HPLC). GC-MS analyses of HPLC fractions able to activate the chimeric receptor identified palmitic acid, oleic acid, linoleic acid, and arachidonic acid as endogenous activators of rPPAR. No other activators were identified. This approach is able to specifically extract and identify endogenous activators of PPAR from a complex biological extract and as such may be valuable in the identification of activators of other orphan receptors in the steroid hormone receptor superfamily. PMID- 8228642 TI - Structure-function relationships of lipoprotein lipase: mutation analysis and mutagenesis of the loop region. AB - The molecular models of two microbial lipases and human pancreatic lipase (PL) have suggested the existence of common structural motifs including a buried active site shielded by an amphipathic surface loop. In an effort to explore the role of residues comprising the loop of lipoprotein lipase (LPL), we have used site-directed mutagenesis to generate three new LPL variants. In variant LPLM1 we deleted 18 amino acids leaving a loop of only 4 residues which resulted in an LPL protein inactive against triolein substrates. In contrast, two other LPL variants with only partial deletions, involving the apical section of the loop [LPLM2 (-8 amino acids) and LPLM3 (-2 amino acids)] manifested normal lipolytic activity. These findings indicate a critical requirement for the maintenance of charge and periodicity in the proximal and distal segments of the LPL loop in normal catalytic function. This is further highlighted by the detection of a mutation in the proximal section of the loop in a patient with LPL deficiency at position 225 which results in a substitution of threonine for isoleucine. The intact catalytic activity of the partial deletion variants (LPLM2 and LPLM3) further suggests that the apical residues of the loop contribute minimally to the functional motifs of the active site. We support this postulate by showing that the conserved glycine in the apical turn section (G229) can be substituted by glutamine, lysine, proline, or threonine without significantly affecting catalytic activity. PMID- 8228643 TI - Presence of platelet-activating factor-acetylhydrolase in milk. AB - Human milk contains numerous factors such as immunoglobulins, lactoferrin, lysozyme, macrophages, etc., which serve an immunoprotective role. Platelet activating factor (PAF) is one of the most proinflammatory agents thus far described. PAF is metabolized to the biologically inactive lysoPAF by the enzyme PAF-acetylhydrolase (PAF-AH). In the present study we have demonstrated that PAF AH activity is present in human milk. The activity was associated with aqueous phase and was not stimulated by the addition of bile salts or Ca2+. The activity of PAF-AH in human milk was not affected by the addition of propranolol or NaCl. PAF, and 1-acyl-2-acetyl-glycerophosphocholine were the only substrates cleaved by the enzyme. Based on these properties it is concluded that the milk PAF-AH is not the lipoprotein or bile salt-stimulated lipase known to be present in milk. Inhibitor studies revealed that the enzyme in human milk was the plasma type PAF AH. The activity of PAF-AH was stable at pH 4.0 at 37 degrees C and the activity varied in milk samples obtained from various species. The enzyme was secreted by milk macrophages. The presence of PAF-AH in human milk may explain, in part, the beneficial effects of breast feeding in the prevention of necrotizing enterocolitis by inactivating the potent proinflammatory autacoid, PAF. PMID- 8228644 TI - Physical behavior of the mixed chain diacylglycerol, 1-stearoyl-2-oleoyl-sn glycerol: difficulties in chain packing produced marked polymorphism. AB - Diacylglycerols (DAG) play an important role in metabolism, signal transduction and protein kinase activation. Naturally occurring DAGs usually contain a saturated chain in the 1-position and an unsaturated chain in the 2-position. We have investigated the physical behavior of 1-stearoyl-2-oleoyl-sn-glycerol (sn SODG) both in the dry and hydrated states by means of differential scanning calorimetry, X-ray diffraction, and NMR. In the dry state the saturated stearate and unsaturated oleate chains have difficulties in packing. As a result marked polymorphism occurs as the chains try to find a suitable packing. Eight phases were found in the dry state: alpha (transition temperature = 16.4 degrees C; delta H = 6.8 kcal/mol); beta 4 (20.7 degrees C; 13.8); beta 3 (21.5 degrees C; 13.8); beta 2 (22.2 degrees C; 14.4); beta 1 (23.1 degrees C; 12.3); beta' (25.7 degrees C; 11.9); gamma 1 (-2.9 degrees C; 0.5), and gamma 2 (-5.9 degrees C; 1.2), all of relatively low stability compared to 1,2 distearoyl-sn-glycerol (beta', 77.2 degrees C; 30.6). gamma 1 and gamma 2 are metastable low temperature phases. beta 1-beta 4 are bilayers (d001 = 3.4, 43.4, 44.7, 46.1 A, respectively) with elements of triclinic parallel chain packing, while beta' is a bilayer (d001 = 47.1 A) with orthorhombic perpendicular chain packing. The metastable alpha phase has hexagonal chain packing and an unusual eight-layer structure (d001 = 174 A). Hydrated sn-SODG contains about one-half of a water molecule per diacylglycerol. Three phases can be distinguished gamma w, alpha w (15.1 degrees C; 6.7) and beta w (19.9 degrees C, 14.3). Both alpha w and beta w are bilayers but alpha w has hexagonal chain packing and beta w is predominantly triclinic parallel packing. Thus, when saturated and unsaturated chains must pack side by side, complex chain conformation, disorder, and instability result giving rise to marked polymorphism. Hydration appears to partly stabilize the interactions. PMID- 8228645 TI - Use of fluorescent cholesteryl ester microemulsions in cholesteryl ester transfer protein assays. AB - In the present report we describe a simple and practical method to assess CETP activity in a defined system by use of microemulsions containing a fluorescent cholesteryl ester analog. The microemulsions are stable, simple to prepare, and can be made to defined composition. Initial transfer rates are easily determined by monitoring changes in fluorescence. We have used the fluorescent cholesteryl ester analog, cholesteryl 4,4-difluoro-5,7-dimethyl-4-boro-3 alpha, 4 alpha-diaza 3-indacenedodecanoate (BODIPY-CE), to demonstrate the utility of this assay. The assay takes advantage of the concentration-dependent self-quenching of BODIPY-CE, when this analog is incorporated into microemulsions. We have used this new assay to demonstrate fluorescent lipid transfer facilitated by rabbit and human d > 1.21 g/ml plasma fraction and recombinant human CETP. A known inhibitory monoclonal antibody (Mab) to human CETP blocked BODIPY-CE transfer in a dose dependent manner. We have also used BODIPY-CE microemulsions to measure CETP activity in whole plasma. PMID- 8228646 TI - Effect of loading frequency on response of the spine to lumbar posteroanterior forces. AB - OBJECTIVE: The purpose of this investigation was to study the spinal deformations produced by applying a force to a lumbar vertebra and to compare the deformation responses at three different loading rates. SETTING: The study was conducted in a laboratory setting using a mechanical device to simulate the manipulative procedure being studied. SUBJECTS: Twelve volunteers (mean age 28.1 yr) formed a convenience sample for testing. All subjects had no significant history of low back pain. INTERVENTIONS: During loading the displacements of the skin were measured at the point of loading, as well as over the spinous processes of L1, T11, T8, and S1 and 30 mm caudad to S1. The loading rates were 1.0 Hz, 0.5 Hz and a very slow "quasistatic" rate. The order of presentation of the loading rates was systematically varied between subjects to minimize the effect of order on the responses. Between 20 and 100 N, the force-displacement relations were highly linear and were characterized by the coefficients of the equation of the line of best fit. MAIN RESULTS: At all three loading rates there were displacements throughout the entire region studied. Changing from quasistatic loading to cyclical loading produced a significant reduction in displacements. CONCLUSIONS: This study found that when a force was applied over L3 there was generalized movement of the whole lumbar and lower thoracic spine. Therefore, a diversity of tissues would be expected to contribute to the resistance to lumbar posteroanterior movements. PMID- 8228647 TI - Comparison of the sacral base angle in females with and without spondylolysis. AB - OBJECTIVE: The aim of this study was to determine if there was a difference in the sacral base angle in females with spondylolysis compared with females without spondylolysis. This is a follow-up study to an earlier pilot program, in which the female sample size was too small. DESIGN: Retrospective study utilizing upright neutral lateral lumbar radiographs of females with evidence of pars defects (test group) and upright neutral lateral radiographs of females with no evidence of pars defects (control group). The control group was age matched to the test group. SETTING: Chiropractic college clinic. INTERVENTIONS: None. MAIN OUTCOME MEASURES: The sacral base angle was measured on each radiograph. RESULTS: The mean sacral base angle was found to be 52.7 +/- 6.8 degrees (mean +/- SD) (n = 22) and 38.6 +/- 7.6 degrees (mean +/- SD) (n = 47) in the test and control groups respectively. This difference was found to be statistically significant (p < .001). CONCLUSIONS: The study has determined that females with spondylolysis have a steeper sacral base angle than females without spondylolysis. It does not, however, indicate whether a larger sacral base angle is a cause or an effect of the spondylolysis. This study also suggests that there is virtually no difference in the sacral base angles between males and females with and without pars defects. PMID- 8228648 TI - The interexaminer reliability of a galvanic skin response instrument. AB - OBJECTIVE: To determine the interexaminer reliability of a protocol of use of a galvanic skin resistance device for detection of low resistance areas along the spinal column, in relatively pain-free subjects. DESIGN: A blinded investigation of concordance of skin resistance examination findings over the spinal column using two clinicians experienced in the use of the instrument. SETTING: A private practice chiropractic outpatient clinic. PATIENTS: Sixty-four male and female chiropractic college students (mean age: 35 yr). The Visual Analog Pain Scale indicated a mean response of 7.6 mm on a 100-mm range. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Concordance of examiners evaluated with the kappa statistic. RESULTS: The results indicated modest levels of concordance for the first study sample (n = 46). The average kappa was 0.37. The second group assessed (n = 18) also demonstrated only modest levels of interexaminer concordance. The average kappa value for this sample was 0.36. CONCLUSION: The use of the Electrical Conductor Scanner instrument for evaluating putative spinal pathology through manifestations in skin resistance in relatively asymptomatic subjects is not supported by the results of this experiment. The unevenness of data generated from this experiment in certain spinal regions necessitates further investigation prior to making any strong conclusions regarding the usefulness of this instrument in the clinical setting. PMID- 8228649 TI - Full-spine radiography: a review. AB - OBJECTIVE: The literature pertaining to full-spine radiography was reviewed to clarify its current role in clinical practice, to identify appropriate parameters for patient selection criteria, to evaluate the validity of spinographic analysis procedures, and to review the latest technological advancements in radiation protection and radiographic technology. DATA SOURCES: The English language literature search included chiropractic textbooks, technique manuals and position papers as well as references from MEDLINE, Chiropractic Research Archives Collection (CRAC) and Chiropractic Literature Analysis and Retrieval System (CHIROLARS). References from non-indexed sources were reviewed from sources obtained in the indexed literature. Current texts on radiographic technology, radiation biology and radiation physics also were reviewed. STUDY SELECTION: The literature was reviewed by a single reviewer. All chiropractic references obtained were referenced. Nonchiropractic references that addressed relevant related topics were also included. DATA EXTRACTION: This study was primarily a qualitative review of the literature. A single observer extracted quantitative data only from controlled studies that examined validity, inter- and intraobserver reliability, and clinical relevance. DATA SYNTHESIS: The conclusions and recommendations in this review are based primarily on the work of experts in the field and well-established principles of radiation biology, radiation protection and radiation physics. CONCLUSIONS: Full-spine radiography and spinographic analysis are controversial procedures in chiropractic. An extensive review of the literature reveals that the role of full-spine radiography is well established. With proper patient selection, careful attention to technical detail and utilization of several technological advancements, full spine radiography is an effective diagnostic and analytic procedure with an acceptable risk/benefit ratio. Although the reliability of certain spinographic parameters has been established, many questions remain regarding the validity and clinical relevance of those parameters. PMID- 8228650 TI - Os odontoideum: chronic neck pain after car accident; failure of two posterior atlantoaxial arthrodeses--medicolegal issues in the occupational setting. AB - OBJECTIVE: To present the case of a woman with previously asymptomatic os odontoideum (OO) who developed chronic neck pain after a car accident. CLINICAL FEATURES: A 32-yr-old woman developed slight constant neck pain following a car accident. Radiographs revealed OO with 12-mm atlantoaxial subluxation on neck flexion. There was no spinal cord compression clinically or on magnetic resonance imaging. INTERVENTION AND OUTCOME: Posterior atlantoaxial arthrodesis was performed twice but the bone graft resorbed for no apparent reason. The patient was treated with analgesics, physiotherapy and a Philadelphia collar to prevent accidental spinal cord compression. CONCLUSION: OO patients may remain asymptomatic if "space available for cord" is sufficient. Even low velocity rotational injury can precipitate chronic neck pain in adults with OO. Posterior atlantoaxial arthrodesis can fail inexplicably even under the best circumstances. Medicolegal issues in the occupational setting are discussed. PMID- 8228651 TI - Automated static perimetry in chiropractic. AB - OBJECTIVE: To demonstrate the value of quantitative static perimetry, in research and practice of spinal manipulation therapy, by reference to the recovery of uniocular loss of vision. CLINICAL FEATURES: A 44-yr-old housewife presented with nonspecific bilateral visual field loss. This visual disability disappeared immediate to pan-spinal manipulation under anesthetic. Later, on review, the presence of a uniocular visual defect was detected by quantitative static perimetry. Further examination revealed no pathology in the eye or brain to explain its occurrence, suggesting that it was due to microischemia of the optic nerve. INTERVENTION AND OUTCOME: The uniocular scotoma recovered immediate to further spinal manipulation under anesthetic, only to recur on two further occasions, each time to disappear immediate to spinal treatment. CONCLUSIONS: This case history demonstrates that spinal manipulation may dissipate microvascular spasm in the brain: even in branches of the carotid arterial system, which is not directly related to the spine. The author feels that the underlying pathology was interictal migraine, which is now known to produce prolonged hypertonic changes in the cerebral microcirculation; and he suggests that spinal manipulation is a treatment to be considered for interictal migraine. PMID- 8228652 TI - Management of cervical disk syndrome utilizing manipulation under anesthesia. PMID- 8228653 TI - Chiropractic magazines. PMID- 8228654 TI - The new Georgia lottery. PMID- 8228655 TI - Dead on arrival. PMID- 8228656 TI - The litigation juggernaut. Part I: The dimensions of the devastation. PMID- 8228657 TI - The key to successful litigation: a strong defense team. PMID- 8228658 TI - When patients cry foul. PMID- 8228659 TI - Who will speak for doctors in the debate over health care? PMID- 8228660 TI - Medical poll-bearers and statistical malpractice. PMID- 8228661 TI - Beneficiary complaints. PMID- 8228662 TI - Attorney survey reveals causes of litigation. PMID- 8228663 TI - Minimizing hospital liability for medical staff physicians. PMID- 8228664 TI - Fear, anxiety, and resistance: those old familiar hallmarks of change. PMID- 8228665 TI - Re: June journal. PMID- 8228666 TI - Re: June journal. PMID- 8228667 TI - Re: Editor's corner, "On guns and violence" (July 1993) PMID- 8228668 TI - The litigation juggernaut. Part II: Strategies and tactics for victory. PMID- 8228669 TI - A country named Sue. PMID- 8228670 TI - Letting people control health care costs. PMID- 8228671 TI - Health system reform: our responsibilities and obligations. PMID- 8228672 TI - Diagnosis and treatment of depression in medical practice. PMID- 8228673 TI - Do the "right" thing. PMID- 8228674 TI - Extracorporeal membrane oxygenation service at Egleston: two years' experience. AB - Extracorporeal membrane oxygenation (ECMO) is a perfusion support procedure that has been used to treat more than 7,000 patients with life threatening cardiac and/or respiratory failure. After 6 months of training and preparation, an ECMO service was opened on January 2, 1991, in Egleston Children's Hospital at Emory University. During the first 2 years, 96 neonatal, 31 pediatric, and 8 cardiac patients have been referred for possible ECMO. Of these 135 patients, 21 had disqualifying conditions. Sixty-four were considered candidates for ECMO but were able to be supported using less invasive therapies; only one of these died. Fifty patients were treated with ECMO of whom 39 survived (78%). Survival rates for neonatal, pediatric, and cardiac cases as separate groups as well as for each diagnostic category within these groups compare favorably with those reported by the international ELSO Registry. Notable in this series is the fact that 26/35 neonatal patients and 7/10 pediatric patients were successfully supported using venovenous (VV) rather than venoarterial (VA) perfusion, with the major indication for venoarterial ECMO being inability to introduce the 14F venovenous catheter into the patient's internal jugular vein. No patient initially managed with VV ECMO required conversion to VA. It is anticipated that avoidance of carotid ligation along with other innovations, such as the impending commercial availability of heparin-coated ECMO circuits, will make ECMO a highly attractive and appropriate therapy for an increasing number of high risk neonatal and pediatric patients in our state and region. PMID- 8228675 TI - God, Profit, and the Newtonian laws of motion: a declaration of independence. PMID- 8228676 TI - Of our uncertainty and our resolve. PMID- 8228677 TI - Preparing for deposition: quality doesn't cost, it pays. PMID- 8228678 TI - A legal labyrinth: lawyer discipline in Georgia. PMID- 8228679 TI - What is this thing called "malpractice"? PMID- 8228680 TI - Medical liability: an ounce of prevention. PMID- 8228681 TI - Alzheimer's disease and dementia. AB - Dementia is frequent in the elderly and, given the expansion of the aged in our society, is a health problem of increasing importance. The most common cause of dementia in the elderly is Alzheimer's disease. Over the past 15 years, great strides have been made in expanding our knowledge of this disease. Some putative risk factors have been defined, the molecular aspects of the pathologic process are being explored, and numerous experimental therapies are being investigated, but to date, clinical studies have demonstrated only mild symptomatic improvement. However, recent advances offer some hope of discovering interventions which might halt or slow the progression of this devastating disease. The clinical evaluation of a patient with dementia should focus primarily on the discovery of treatable causes or contributing factors which may be ameliorated. The primary care physician plays an important role in diagnosis, in management of psychiatric complications, and in providing guidance and counsel on psychosocial and legal issues. PMID- 8228682 TI - Moral authority, moral standing, and moral controversy. PMID- 8228683 TI - Persons and death: what's metaphysically wrong with our current statutory definition of death? AB - This paper challenges the recommendations of 1981 President's Commission for the Study of Ethical Problems in Medicine and Biomedical and Behavioral Research that all jurisdictions in the United States should adopt the Uniform Determination of Death Act, which endorses a whole-brain, rather than a higher-brain, definition of death. I argue that the Commission was wrong to reject the "personhood argument" for the higher-brain definition on the grounds that there is no consensus among philosophers or the general population as to what constitutes "personhood". I claim that philosophers agree that some potential for cognitive function is necessary for personhood and that, when this is absent in cases of anencephaly and persistent vegetative state (PVS), the individual should be considered dead. I further argue that the lack of consensus among the general population is due in large measure to misunderstandings about the medical reality of PVS and beliefs influenced by feelings for a specific individual in PVS. I also examine and reject two tutorist arguments which have been used to support the Commission's position: that the higher-brain definition would threaten the severely senile and severely retarded, and that there are not currently adequate medical techniques for determining when all higher-brain activities have ceased. PMID- 8228684 TI - Deficiencies in the National Institute of Health's guidelines for the care and protection of laboratory animals. AB - This paper is a critique of NIH guidelines for the care and protection of laboratory animals. It exposes four serious deficiencies in these guidelines: (1) failure to make it clear that the mere pursuit of knowledge does not justify using animals; (2) failure to give any guidance concerning what constitutes human benefit or well-being; (3) failure to countenance trade-offs between human benefit or well-being and animal well-being; (4) failure to clearly specify what constitutes keeping animals in an 'environment appropriate to the species and its life history.' It concludes with the suggestion that the construction and revision of these guidelines is too important to be left to the professionals. PMID- 8228685 TI - Economic perspectives on bioethics. AB - Wendell Stephenson argues in this issue of The Journal that the National Institutes of Health's standards for the treatment of laboratory animals fail to give any guidance concerning human well-being nor do they balance human well being and animal well-being. Stephenson fails, however, to demonstrate how such a balance is to be known. In arguing for reform he implies greater state control without showing that such control would improve the situation. Indeed there are good reasons to think that such control may be harmful. PMID- 8228686 TI - The personhood of the human embryo. AB - My interlocutor is anyone who denies personhood to the embryo on the grounds that a human person can exist only in conscious activity and that in the absence of consciousness a person cannot exist at all. I probe personal consciousness to the point at which the distinction between the being and the consciousness of the human person appears, and argue on the basis of this distinction that the being of a person can exist in the absence of any consciousness. I proceed to argue that it is not only entirely possible for the embryo to be a human person, but that, given the embodied personhood of us human beings, this is the only reasonable assumption which we can make. PMID- 8228687 TI - Personhood, moral strangers, and the evil of abortion: the painful experience of post-modernity. AB - The epistemological and sociological consequences of post-modernity include the inability to show moral strangers, in terms they can see as binding, the moral wrongness of activities such as abortion. Such activities can be perceived as morally disordered within a content-full moral narrative, but not outside of the context it brings. Though one can salvage something of the Enlightenment project of justifying a morality that can bind moral strangers, one is left with moral and metaphysical views that can be recognized as impoverished and incomplete by those who live their lives within the embrace of a content-full moral narrative. The cardinal dualism of post-modernity is not that which separates mind from body, but the gulf between the morality binding moral strangers and that binding moral friends. PMID- 8228688 TI - Mechanical and non-mechanical ventilation of respiratory failure in chronic obstructive pulmonary disease. AB - Among 182 episodes with ARF (PaCO2 > 50 torr) in 400 episodes of COPD patients who were admitted to Chulalongkorn Hospital during the period 1982 to 1986, despite conservative treatment, 66 developed severe acute respiratory failure requiring assisted ventilation. Patients with a history of chronic cough, pneumonia as a precipitating factor and more severe ARF on admission, as indicated by palpitation, headache, cyanosis, alteration of consciousness, cor pulmonale and decompensated acidosis (pH < 7.30), were likely to require mechanical ventilation. Indications for mechanical ventilation were carbon dioxide narcosis (43 episodes), severe hypoxemia despite on a high FIO2 (one episode), various combination parameters of respiratory muscle fatigue, cardiovascular instability (22 episodes). The major complications of mechanical ventilation were pneumonia, sepsis, pneumothorax, UGI bleeding of 16, 8, 5 and 9 episodes, respectively. The average duration of assisted ventilation and hospitalization were 15.8 and 19.02 days, respectively. The mortality rate was 50 per cent in the mechanical ventilation group compared with 9.8 per cent in the non-mechanical ventilation group. Increased mortality rate was found in those with pneumonia as the precipitating factor (68.4 vs 14.3%, respectively, in comparing the two groups). Complications of mechanical ventilation, which included pneumonia, sepsis, fluid overload, hyponatremia and persistent acidosis, were high-risk factors for the non-surviving group. PMID- 8228689 TI - Collodion baby: the out-come of long-term follow-up. AB - About 10 Thai collodion babies were treated at the Department of Pediatrics, Faculty of Medicine, Chulalongkorn University from 1970 to 1990. Most of them were premature and small for gestational age infants. The sex incidence of males to female was 3:2. Only half of the cases had a family history of ichthyosis. The severity of thickness and tightness of the collodion membranes varied from mild to severe. The clinical and histopathologic findings revealed 5 cases of lamellar ichthyosis (50%), of which one case had episodes of pustular psoriasis. One case evolved to be ichthyosis vulgaris 10 per cent, one case had palmoplantar hyperkeratosis (10%), one case had been apparently normal up to 18 years old, one died of respiratory infection and another case was lost from follow-up. All cases of lamellar ichthyosis were associated with mental retardation with some degree of slow physical development. One severe collodion baby suffered from congenital absence of distal phalanges of toes with increased thickness of the finger nails. PMID- 8228690 TI - Posterior sagittal anorectoplasty for imperforate anus: a preliminary result. AB - Posterior Sagittal Anorectoplasty (PSARP) is a new technique which was pioneered by Pena and de Vries in 1982 for the repair of high anorectal malformations. It is based upon complete exposure of the anorectal region by means of a median sagittal incision that runs from the sacrum to the anal dimple cutting all muscle structures behind the rectum. The incision is kept in the median plane using a muscle or nerve stimulator as a guide. Between March 1988 and April 1989, we operated on 10 patients who had high and intermediate types of imperforate anus according to the "Wingspread Classification". All of the patients had PSARP as the definitive operation. The age of the patients were between 1.5 months and 15 years at the time of operation, four were under six months. Nine patients had this operation as the primary procedure, one patient had re-pullthrough after failure to gain continence by a previous conventional abdomino-perineal pullthrough operation. Six out of 8 patient had good results, two patients had fair results according to Kiesewetter's clinical criterion for fecal continence. Two patients are waiting for colostomy closure and are yet to be evaluated. Two of the good results included a 15-year-old girl who had high type with rectovaginal fistula, and a 2-month-old male infant with a recto-urethral fistula. In a re-pullthrough patient, the continence, as evaluated by loopogram and defecogram through the colostomy before closure of the colostomy, is good.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228691 TI - An analytic cross sectional study of anemia in rural area, Amphoe Phon, Khon Kaen. AB - Anemia is a major public health problem in Thailand with the estimate prevalence of 25 per cent. The last survey of the prevalence of anemia was done in 1979, so we performed another survey to observe the trend of change in prevalence of anemia in Khon Kaen. We conducted a community survey by multistage random sampling. In a population aged 30-65 years in 13 villages in Amphoe Phon. The mean hematocrit level was 40.1 +/- 5.1 per cent (42.8 +/- 4.6% in males, 38.1 +/- 4.5% in females). The prevalence of anemia (Hct < 39% in males, < 36% in females) was 22.2 per cent (16.4% in males, 26.4% in females). The mean level of hematocrit of patients admitted for elective surgery in Srinagarind Hospital was 38.6 +/- 5.5 per cent which was significantly lower than in the rural area (P < 0.001) and the prevalence of anemia was 25.9 per cent. The prevalence of anemia increased with age and decreased with high education and body mass index. Comparing to the data in 1979, the prevalence in males increased from 13.9 to 16.4 per cent and from 23.3 to 26.4 per cent in females. The prevalence of anemia in Khon Kaen is still high and has increased compared to the data in 1979, and thus, remains a major public health problem. PMID- 8228692 TI - Sequential intravenous/oral ciprofloxacin in the treatment of severe multiresistant gram-negative infections. AB - Twenty-one patients with severe multiresistant gram-negative bacillary infections were treated with ciprofloxacin, intravenously followed by oral. The mean duration of therapy was 13 days. Causative organisms were Klebsiella pneumoniae (18 patients), Pseudomonas aeruginosa (2) and Salmonella enteritidis (1). The overall clinical improvement was 85 per cent, with a bacteriologic improvement of 90 per cent. Three patients died, one had fungemia, another had persistent bacteremia, and a third had progressive lung infiltration despite eradication of bacteremia. Superinfections occurred in 2 patients, and the other 2 had colonization of the wounds. It is shown that this treatment is effective and safe for the treatment of severe multiresistant gram-negative infections. PMID- 8228693 TI - Detection of human immunodeficiency virus type 1 (HIV-1) proviral DNA in breast milk and colostrum of seropositive mothers. AB - There is increasing evidence of vertical transmission of HIV-1 to infants through breast feeding of milk from HIV-1 infected mothers. It has been postulated that transmission occurs mainly via ingestion of infected cells in breast milk and colostrum. In this study, detection of HIV-1 proviral DNA was used to prove that cells from colostrum and milk do contain HIV. DNA were extracted from these cells of colostrum and milk of 18 seropositive mothers and amplified by nested PCR for HIV-1 gag and pol and 44 per cent were positive mostly by two primers. All ten negative control samples from seronegative mothers were negative. This study demonstrated the infectivity of breast milk and colostrum. Nevertheless, recommendation against breast-feeding should be weighed against poor alternatives in low socioeconomic families. PMID- 8228694 TI - Knowledge and attitudes about AIDS: a comparative study of Thais involved in sexual occupations, university students and immigrants living in the United States. AB - Knowledge about AIDS among: (a) 89 individuals involved in prostitution related occupations, (b) 465 university students, and (c) 220 Thais living in the United States was assessed to further evaluate the efficacy of Thailand's AIDS educational programs. While the average number of items answered correctly by the three groups were not equivalent (p < 0.001), all participants possessed substantial factual AIDS knowledge. As predicted, those involved in prostitution had the least accurate information (92% correct). Contrary to initial hypothesizing, however, AIDS knowledge was identical (95%) for the samples of university students living in Thailand and Thais who have been exposed to intensive AIDS information campaigns in the United States. Taken together, these findings suggest that Thailand's AIDS educational efforts have achieved considerable success. Television, newspapers, and radio provided information for most participants while a few learned about AIDS from their physician or in educational institutions. A majority expressed interest in improving their understanding about AIDS. The extent of infection among prostitutes appears to be high with nearly one-quarter of those in the sexual occupations group reporting that they knew someone with the disease. Techniques to better inform prostitutes about AIDS are discussed and a common outcome instrument to measure AIDS knowledge and attitudes is recommended. PMID- 8228695 TI - Hereditary coproporphyria: a case report. AB - A case of hereditary coproporphyria was reported, he was a 21-year-old farmer, presenting with abdominal pain and fever. His manifestations were composed of all classical symptoms of acute hepatic porphyrias i.e. convulsions, psychosis, hypertension and respiratory failure as well as dark red urine with positive Watson-Schwartz test. Because of lack of cutaneous photosensitivity and strikingly increased urinary coproporphyrin, diagnosis of hereditary coproporphyria was most likely. Precipitating factor could not be identified. He responded well to glucose and other symptomatic treatment during the first admission but not in the second. He died from respiratory failure. PMID- 8228696 TI - Precipitous hypotension after nifedipine in pheochromocytoma: report of 2 cases. AB - Two patients with pheochromocytoma initially responded to oral or sublingual nifedipine with marked reduction in systolic blood pressure (range 50-160 mmHg) without concomitant increment in the heart rate or clinical consequences. This could possibly be due to the combination of hypovolemia and negative inotropism. PMID- 8228697 TI - Hemodynamic effects of a beta adrenergic blocking agent-atenolol in patients with severe mitral stenosis. AB - The hemodynamic effects of atenolol, a cardioselective adrenergic blocking drug, were studied in 10 patients with clinically severe mitral stenosis during cardiac catheterization to evaluate the severity of disease before cardiac surgery. At 5 minutes after intravenous injection of 0.1 mg/kg of atenolol, there were significant reductions of heart rate, cardiac index, pulmonary pressure, pulmonary capillary wedge pressure and mitral valve gradient. The right and left ventricular filling pressures were increased slightly. The pulmonary and systemic vascular resistance did not change after atenolol. The PR interval although prolonged significantly was still within normal range. No serious side effects occurred during the post-catheterization 24 hour observation period except for one patient who developed slow ventricular rate (48/min) during the study. The hemodynamic effects of atenolol observed in this study appear to benefit the patients with mitral stenosis by reduction of pulmonary pressure, pulmonary capillary wedge pressure and mitral valve gradient. These hemodynamic changes may, therefore, lead to improvement of pulmonary congestive symptoms. PMID- 8228698 TI - A prevalence survey of lipids abnormalities of rural area in Amphoe Phon, Khon Kaen. AB - A population survey in 13 villages and 3 sub-districts of a rural area in Amphoe Phon, Khon Kaen showed that mean cholesterol and high density lipoprotein cholesterol (HDL-C) level were 163.0 +/- 35.3 and 29.6 +/- 9.1 mg/dl respectively which is very low compared to Bangkok and other affluent countries. The cholesterol level in females was higher than in males (male 158.0 +/- 33.2, female 166.6 +/- 36.4) and the HDL-C level in males was higher (30.2 +/- 9.0 vs 29.1 +/- 9.2). The cholesterol level increased with age and obesity while HDL-C decreased with obesity. The prevalence of hypercholesterolemia and low HDL-C was 3.0 and 74.9 per cent respectively. The diet of the survey population was high in carbohydrate (80% of calories) and fat intake was very low (8% of calories). The factors which were associated with high cholesterol levels were age, female sex, obesity and increased fat intake while those of low HDL-C were obesity and being a farmer. The low fat intake may explain for the low total cholesterol, HDL-C HDL C/cholesterol ratio observed in our population compared to other studies. PMID- 8228699 TI - Blood lipids, the risk factor of cerebral infarction. AB - A hospital-based case-control study was undertaken at Prasat Neurological Hospital and Institute, Bangkok, in 1990 to determine the association between blood lipid variables (i.e., total cholesterol and triglycerides) and cerebral infarction. To alleviate the unwanted effects of sample selection a number of inclusion and exclusion criteria were also made. A total of 193 newly diagnosed cerebral infarction patients together with an equal number of age-sex-matched controls, male and female 45 to 84 years of age, were studied. The association was assessed by logistic regression analysis. Other potential risk factors for cerebral infarction were entered in the logistic models as covariates to be adjusted. The results showed that, total cholesterol and triglycerides were statistically significant related to cerebral infarction (p < 0.0001) in univariated analysis. After adjusting for the covariates through multivariate logistic regression analyses, total cholesterol remained significantly related to cerebral infarction (p = 0.0430) but triglycerides was not significant (p = 0.3953). Other factors, namely, hypertension, cardiac impairment, diabetes mellitus and hemoglobin concentration were strongly related to cerebral infarction in both the univariate and multivariate analyses. The interaction between blood lipids variables and potential risk factors was also assessed. The finding showed significant multiplicative interaction between triglyceride levels and diabetes mellitus status (p = 0.0026). Other interaction terms did not approach statistical significance. PMID- 8228700 TI - Intestinal anisakiasis: first reported case in Thailand. AB - A case of intestinal anisakiasis is reported. The patient came with the symptoms of acute abdominal obstruction. The diagnosis was obtained by identification of the parasite in the tissue sections of the resected segment of the small intestine. This case appears to be the first reported case in Thailand. PMID- 8228701 TI - Clinical comparison of patients with ankylosing spondylitis, Reiter's syndrome and psoriatic arthritis. AB - Between 1976 and 1989, 160 cases of seronegative spondyloarthropathy (SNSA) were admitted to the Department of Medicine at Chulalongkorn Hospital. The prevalence of idiopathic ankylosing spondylitis (IdAS) was 52 cases (32.5%), Reiter's syndrome (RS) 68 cases (42.5%), psoriatic arthritis (PsA) 28 cases (17.5%), reactive arthritis eight cases (5.0%) and Behcet's disease four cases (2.5%). Clinical comparison of the patients with IdAS, RS and PsA showed a male predominance in IdAS (90.2%), RS (97.1%) and PsA (71.4%). There was a significant difference (p < 0.01) between IdAS and PsA, and RS and PsA. The initial articular manifestation usually occurred in the younger age group (IdAS, 22.15; RS, 22.91; and PsA, 30.86 years); however, there was a significant difference (p < 0.05) between IdAS and PsA, and RS and PsA. Initial peripheral arthritis was found in IdAS (51.9%), RS (91.2%) and PsA (92.6%); there was a significant difference (p < 0.001) between IdAS and RS, and IdAS and PsA. The symptom of back pain was found in IdAS (78.8%), RS (38.2%) and PsA (21.4%); there was a significant difference (p < 0.001) between IdAS and RS, and IdAS and PsA. During physical examination, peripheral arthritis was evident in the IdAS (42.2%), RS (88.2%) and PsA (92.2%) patients; likewise sacroiliitis was found in the IdAS (100%) RS (54.4%), and PsA (57.2%) patients. Evidence of ankylosing spondylitis was found in the IdAS (100%), RS (22.1%) and PsA (46.4%) patients. These findings show a significant difference (p < 0.001) between patients with IdAS and RS, IdAS and PsA. Other associated symptoms were similar, particularly evidence of enthesopathy (tendonitis, heel pain, plantar fasciitis), the polyarticular pattern was more common than the mono-articular pattern. Hip joint was significantly (p < 0.05) more commonly involved in patients with IdAS than in those with RS and PsA. Associated symptoms, particularly genital lesion or skin lesion, are specific symptoms for RS and PsA, respectively. PMID- 8228702 TI - Vitamin B12, folic acid and haematological status in elderly Thais. AB - Serum vitamin B12, folic acid and haematological data from 147 elderly people (55 males and 92 females) who visited the special clinic for the elderly at Rajvithi Hospital, Bangkok between July and November 1989 were investigated. The individuals studied came from a health-conscious group of the middle socio economic class in Bangkok. All of them were fairly well except for minor ailments and typical diseases of elderly people such as hypertension, mild to moderate degree coronary heart diseases and non-insulin dependent diabetes mellitus. There was a statistically significant difference in haemoglobin concentrations between males and females. According to the standard haemoglobin cut-off point values of 13 g/dl for males and 12 g/dl for females, anaemia was detected in 22 (15%) of the 147 subjects. The percentage of folic acid deficiency was found to be 20.6 per cent (30 of the 147 cases). Vitamin B12 insufficiency was found in only 6.9 per cent (10 of the 147 cases). No statistically significant correlation between haemoglobin, folic acid and vitamin B12 was found. However, when the data were grouped according to different intervals of increasing haemoglobin concentrations, for females there was a tendency for serum vitamin B12 to decrease, and serum folic acid to increase in both males and females. The results of this study suggest that folate deficiency may play a role in the occurrence of anaemia in elderly people, and therefore, dietary counselling and supplementation of folic acid are recommended. PMID- 8228703 TI - Ultrasonic fetal transverse trunk diameter in normal northern Thai women. AB - Fetal transverse trunk diameter (TTD) were measured by two obstetric sonographers, using Aloka model SSD 630 and 650. The measurements were performed on 1,009 occasions in 235 women during normal pregnancies between the 14th and 40th weeks (age range 17-37 years) (4.5 measurements per pregnancy, on average). The relation between TTD and menstrual age was determined and mathematical modeling of the data demonstrated that the linear quadratic function was optimal (r2 = 0.973). The most fitted regression equation was obtained by regression analysis. 1.) TTD = -2.69055 + 0.38326 G.A - 0.00140137 GA2 2.) GA = 7.222585 + 2.82632 TTD + 0.02605 TTD2. Interestingly, TTD value for each gestational week in this study is not so different from that of the European data. Predicted TTD values for a given gestational week were determined and presented in tabular form. The values from this study may be important baseline data for evaluation of fetal growth in our population. PMID- 8228704 TI - Clinical and histopathological features of secondary syphilis. AB - Fifty-four patients with secondary syphilis were studied in regard to the clinical manifestations, response to treatment and histopathology of the skin lesions. The correlation between the skin lesions and histopathology, between the duration of skin lesions and VDRL titer, between type of skin lesions and VDRL titer were also determined. The clinical manifestations varied from macular, maculopapular, papular, papulosquamous and urticarial lesions. The hair loss usually occurred on the scalp, but the eyebrows or even total body hair loss could occur. The response to treatment was good, only one patient relapsed. The histopathology was related to the clinical manifestations, there was sparse inflammatory cell infiltration in the dermis in macular lesions, but more dense infiltration as well as more epidermal change in papular and papulosquamous lesions. There were significant correlation between the duration of skin lesions and VDRL titer, but no correlation between types of skin lesions and VDRL titer. PMID- 8228705 TI - Growth and development of infants conceived by in vitro fertilization: a preliminary report. AB - This is a preliminary report on the first 24 babies born during the period of 1 year follow-up after IVF-ET in Chulalongkorn Hospital. Based on this study, our data suggests that these children have a normal range of growth and development. There was also no evidence of major congenital anomalies identified in spite of some minor ones. However, a need for more detail and on-going long-term follow-up at least until school age is being planned. PMID- 8228706 TI - Chemical components of gallstone in southern Thailand. AB - The gallstones from 55 patients in Southern Thailand were analyzed to disclose the chemical components using infrared grating spectrophotometry. The main components were calcium bilirubinate, calcium phosphate, calcium carbonate, choleterol and protein inproportions 52.6, 5.5, 5.5, 30.9 and 5.5 per cent of stones, respectively. These proportions are not different from those of gallstones in the northern or central regions of Thailand. PMID- 8228707 TI - Situation of hypertension in some Bangkok slums. AB - A cross-sectional epidemiologic survey was undertaken in slums in the catchment areas of two health centers in Bangkok. Two hundred and sixty representative households were randomly selected. All adults (993) aged 20 years and over who lived in the households were asked to participate. The results were, however, based on 809 subjects (89%) who responded and whose data were completed. It was found that prevalence of hypertension (blood pressure > or = 160/95 mmHg) was 14 per cent with a 95 per cent confidence interval of 11.8 and 16.4 per cent. Nearly half of the hypertensives were found to be mild cases (diastolic blood pressure = 95-105 mmHg). Less than half of the hypertensives were aware of their condition. Even a lower proportion was under treatment and had their blood pressure controlled. The results confirmed earlier studies that slum inhabitants seem to be at high risk of hypertension. Furthermore, the status of hypertensives in terms of awareness, treatment and control was unacceptably poor. This prompts a need to develop community programmes to cope with the problems. PMID- 8228708 TI - Community- and hospital-acquired pneumonia in medical intensive care unit. AB - Seventy-four cases of pneumonia admitted to the Medical Intensive Care Unit of Chulalongkorn Hospital from January 1986 to December 1988 were analyzed. There were 52 males and 22 females with 46, 26 and 2 cases of community-, hospital- and combined-acquired pneumonia, respectively; the average age on admission was 58 +/ 18.1 years. The majority of the patients had abnormal host or underlying pulmonary diseases. Most of the cases with community-acquired pneumonia were admitted to the ICU because of respiratory failure (85%) and most of the cases with hospital-acquired pneumonia had ventilator-associated pneumonia (71.4%). The dominating pathogens in community- and hospital-acquired pneumonia were Gram positive and Gram-negative organisms, respectively. There were no significant differences in initial clinical manifestations, laboratory data, complications, between the two groups. However, the duration on antibiotic, respirator, leukocytosis and stay in hospital were significantly longer in hospital acquired than community acquired pneumonias. Overall mortality was 63.5 per cent, the causes of which were progressive or uncontrolled pneumonia and mechanical ventilation complications in 37.8 and 25.7 per cent of the cases, respectively. High-risk factors of the non-surviving groups included pneumonia on assisted ventilation, mechanical ventilation complications, shock and the presence of Gram negative bacilli. Our study indicates that the patients required assisted ventilation because of the severity of the underlying pulmonary conditions rather than the extent of the pneumonia itself. Gram-negative pneumonia predisposed the patients requiring mechanical ventilation to develop complications. The duration on mechanical ventilation, ICU admission and hospitalization were 13.84, 13.21 and 29.08 days, respectively. PMID- 8228709 TI - Nutritional health and parasitic infection of rural Thai women of the child bearing age. AB - The nutritional and health status of non-pregnant women of child bearing age from 20 villages of two districts in Maha Sarakham Province, approximately 500 km northeast of Bangkok, were investigated in April 1987. About 12 per cent of the studied women had a body mass index below 18.7 which is used as a cut-off point of being undernourished. Fat stores and muscle mass were smaller when compared to western females. Only two per cent of the women investigated had low serum albumin, indicating a severe deprivation in the nutritional status. A relatively high prevalence of anaemia was also found. Iron deficiency and haemoglobinopathy are common causes of anaemia. In addition, vitamin B2 depletion also contributes to the high rate of anaemia. Gastrointestinal parasitic infection rates were high with liver fluke, hookworm and echinostomiasis. Hookworm infection had no effect on the rate of anaemic, most probably the worm load in the women infected was rather low. It is recommended that the nutritional health of rural adolescents and young women should be enhanced by generally improving dietary habits and the quality of nutrient intake especially through protein, vitamins and micronutrient. Public health programmes focused on preventive activities should be aimed at this group in order to reduce the prevalent rate of undernutrition, anaemia and parasitic infection. PMID- 8228710 TI - Cervical cancer incidence in the south of Thailand 1988-1989. AB - In order to assess the incidence of cervical cancer in Southern Thailand, the data of all diagnosed cases at or above the level of the community hospital in each province were collected during the period January 1, 1988 to December 31, 1989. The crude annual incidence was 9.13 and 8.67 per 100,000 women in 1988 and 1989 respectively. The age-standardized annual incidence in these years was 13.42 and 13.07 per 100,000. Phuket had the highest incidence and Narathiwat the lowest in both years. In upper and lower subregions the crude annual incidence was 7.36 and 10.76 in 1988 and 8.95 and 8.41 in 1989. These rates are slightly lower than the incidence in Khon Kaen province collected between 1985 and 1987 from a population-based cancer registry. PMID- 8228711 TI - Ultrasonic measurement of the fetal head to abdominal circumference ratio in normal pregnancy. AB - The relation between fetal HC/AC ratio and menstrual age was determined from 14th 40th weeks gestations by analysis of 1,211 measurements examined with real-time ultrasound scanner Aloka 630, 650. The mean HC/AC ratio with +/- 2SD and percentile ranks were determined and presented in both graphic and tabular forms. The mean ratio was 1.207 at 14 week's but decreased slowly until 30 week's when the ratio was 1.110 thereafter there was a rather sharp fall in the mean ratio 1.010 at 36 weeks and 0.967 at 40 weeks and then the variability decreases. Interestingly, HC/AC ratio value for each gestational week in this study was not so different from those of western studies. We hope that the value from this large series is important baseline data for evaluation of fetal growth in our population and is more appropriate for application with Thai women than caucasian data. PMID- 8228712 TI - Biliary stent: a report of experience in 25 Thai patients. AB - Biliary stent insertion was done in 25 patients. Eighteen were successful, 14 by endoscopic method and 4 by combined endoscopic percutaneous method. Twelve with malignant diseases and 4 with benign diseases had satisfactory drainage. Failed stent insertion occurred in 7 cases. Stent malfunction was found in 30 per cent and occlusion was the most common problem. Complications occurred in 32 per cent and most were mild. The 4 per cent mortality of the procedure is acceptable. Biliary stent should be considered as an alternative or as an adjunctive treatment to surgery. PMID- 8228713 TI - An efficacy study of ampicillin versus cefazolin prophylaxis in patients undergoing cesarean section. PMID- 8228714 TI - Prenatal sonographic diagnosis of meconium peritonitis: a case report. AB - A case of meconium peritonitis that was diagnosed ultrasonographically in the third trimester is presented. Fetal ascites, intra-abdominal calcification, left communicating hydrocele, and polyhydramnios were detected on antenatal ultrasonography. Specks of calcification were also demonstrated on abdominal radiography postnatally. Laparotomy confirmed the diagnosis of perforated terminal ileum with meconium peritonitis. The obstetric and neonatal implications of meconium peritonitis are discussed with literature review. PMID- 8228715 TI - A computer heart model incorporating anisotropic propagation. I. Model construction and simulation of normal activation. AB - Present-day computer models of the entire heart, capable of simulating the activation isochrones and subsequently the body surface potentials, focus on considerations of myocardial anisotropy. Myocardial anisotropy enters into play at two levels, first by affecting the spatial pattern of activation owing to faster propagation along cardiac fibers and second by altering the equivalent dipole sources used to calculate the surface potentials. The construction of a new and detailed model of the human heart is described, based on 132 transverse sections obtained following a computed tomography scan of a frozen human heart whose chambers were inflated with pressurized air. The entire heart anatomy was reconstructed as a three-dimensional array of approximately 250,000 points spaced 1 mm apart. Conduction in the thin-walled atria was assumed isotropic from the sinus node region to the atrioventricular node, where it was subject to a 50 ms delay. A two-tier representation of the specialized conduction system was used, with the initial segments of the left and right bundles represented by a system of cables that feeds to the second tier, which is a sheet of conduction tissue representing the distal Purkinje system. Approximately 1,120 "Purkinje myocardium" junctions present at the terminations of the cables and sprinkled uniformly over the sheet, transmit the excitation to the ventricles. A stylized representation of myocardial fiber rotation was incorporated into the ventricles and the local fiber direction at each model point used to compute the velocity of propagation to its nearest neighbors. Accordingly, the activation times of the entire ventricular myocardium could be determined using the 1,120 or so Purkinje myocardium junctions as start points. While myocardial anisotropy was considered in the ventricular propagation process, it was ignored in the computation of the equivalent dipole sources. Nevertheless, the computed electrocardiogram, vectorcardiogram, and body surface potential maps obtained with the new heart model properly positioned inside an inhomogeneous torso model were all within normal limits. PMID- 8228716 TI - A computer heart model incorporating anisotropic propagation. II. Simulations of conduction block. AB - This study describes the simulation of the more common types of conduction blocks with a computer model of the heart incorporating anisotropic propagation. The rationale was to test the model as to its ability to simulate these blocks by physiologically justifiable adjustments of the conduction system alone. The complete blocks were generated by simply blocking conduction totally at selected sites in the proximal conduction system, and the incomplete blocks by slowing down the conduction velocity in the proximal system. Also simulated were the left fascicular blocks and the bilateral blocks. All simulated electrocardiograms, vectorcardiograms, body surface potential maps, and epicardial isochrones for these blocks were similar to clinically observed data, with the exception of the left posterior hemiblock, which was slightly atypical. This could be because such blocks are usually accompanied by other cardiac pathologies not included in our simulations. The model also supports van Dam's observation that during left bundle branch block the passage of activation from right to left occurs via slow myocardial activation with no evidence of a local delay due to a septal barrier. Finally, the model suggests that a left bundle branch block with a normal frontal plane QRS axis may simply represent a case of an incomplete left bundle block, whereas one that exhibits a left axis QRS deviation in the frontal plane represents a more severe complete left bundle branch block. PMID- 8228717 TI - Assessment of reperfusion in myocardial infarction by body surface electrocardiographic mapping. AB - To determine the efficacy of body surface potential mapping to detect and quantify reperfusion in acute infarction, 66 patients were studied by repeated body surface potential mapping before and after administration of the thrombolytic agent. The QRS and ST-segment were analyzed and compared to the arterial patency as assessed by arteriography within 10 days. The infarct-related vessel was patent in 50 patients and occluded in the remaining 16. In 6 of the 15 patients in whom thrombolytic therapy was started within 2 hours of the onset of chest pain the ST-segment changed from that of an acute infarction pattern to that of a normal pattern, and the QRS pattern either remained normal or recovered prior to discharge. In two additional patients the QRS pattern returned to normal prior to discharge from the hospital. In the 51 patients with later thrombolytic therapy (> or = 2 hours) the degree of ST elevation and depression decreased more than either the control infarction group (36 inferior and 73 anterior patients) or the group in whom reperfusion attempts were unsuccessful, but the pattern of the map remained that of an infarction. The QRS maps showed that in the first 48 hours recovery of potential was insufficient to distinguish those with successful thrombolysis. Early reperfusion could be detected by body surface potential mapping and the eventual damage predicted from the degree of change in the QRS map. Later reperfusion could be surmised but not quantified. PMID- 8228718 TI - Electrical current paths in acute pericarditis. AB - The electrocardiographic changes accompanying pericarditis consist of ST elevation in most of the leads of the 12-lead electrocardiogram. The source of this ST elevation is thought to be local inflammatory changes in the epicardium underlying the inflamed pericardium. The current from this area of ST elevation must return to some unaffected region of the heart and this should be associated with a region of ST depression. This current path from the external epicardial surface has been postulated to flow back into the endocardium through the great vessels and atria. To test this hypothesis, 18 patients with pericarditis were studied by body surface potential mapping and inverse epicardial potential distributions were computed. The resultant maps were compared to those of normal people and patients with acute anterior infraction. Epicardial maps from patients with pericarditis showed a region of current flow into the heart over the great vessels and atria in all 18 patients. This pattern was not seen in normal patients or infarction patients and was consistent with the mechanism resulting in ST elevation in pericarditis being one of current flowing from the epicardium out into the thorax and back into the heart through the great vessels and atria. PMID- 8228719 TI - Body surface potential maps in patients with familial amyloid polyneuropathy. AB - The purpose of this study was to evaluate the characteristics of body surface potential maps in patients with cardiac amyloidosis. The study population consisted of 30 patients with familial amyloid polyneuropathy and 50 age-matched normal volunteers. The patients were classified into one of the following three stages: stage I, peripheral neuropathy limited to the lower limbs; stage II, neuropathy involving both the lower and upper limbs; and stage III, bedridden because of extensive progressive neuropathy. Electrodes for the body surface potential maps were placed at 87 points (59 anterior and 28 posterior) on the chest. To analyze these body surface electrocardiograms, isopotential maps, isochrone maps, and isointegral maps were used. The mean values of the positive potential were significantly lower in the advanced stage (1.9 +/- 0.2 mV in stage I, 1.0 +/- 0.2 mV in stage II, and 0.7 +/- 0.2 mV in stage III). Prolongation of ventricular activation time was observed on the anterior and lateral chest. The mean QRST isointegral maps of the patients in the advanced stage of cardiac amyloidosis showed a large negative area over the anterior and left lateral chest, the positive areas were small and their potentials were very low. In addition, 18 (60%) of the 30 patients had a multipolar pattern in the QRST isointegral maps. The changes of the body surface potential maps correlated with clinical staging and echocardiographic findings. PMID- 8228720 TI - Wavelet analysis of high-resolution signal-averaged ECGs in postinfarction patients. AB - The authors present an original method for the discrimination of patients prone to ventricular tachycardia. The wavelet transform, which is a new time-scale technique suitable for transient signal detection, was applied to bipolar unfiltered X, Y, Z signal-averaged electrocardiograms in 20 postinfarction patients with sustained ventricular tachycardia, in 20 myocardial infarction patients without ventricular tachycardia, and in 10 healthy subjects. An improved automated algorithm for the detection and localization of sharp variations of the signal, based on coherent detection of the local maxima of the wavelet transform, was developed. A risk stratification method, based on the detection of at least one singularity at or after a point defined with reference to the QRS onset, was assessed. The optimum cutoff point, found 98 ms after the onset of QRS, provides a specificity of 90% and a sensitivity of 85%. The authors conclude that wavelet analysis makes it possible, in this group of patients, to discriminate those with ventricular tachycardia. It yields better results than those obtained from the conventional time-domain approach. PMID- 8228721 TI - Automated measurement of QT interval dispersion from hard-copy ECGs. AB - Increased "dispersion" of the QT interval of the electrocardiogram has been proposed as a marker for increased risk of cardiac arrhythmias, but definitive identification of its independent predictive value requires accurate and reproducible measurement in large numbers of cases. A personal computer-based technique for (1) converting hard-copy electrocardiograms to digital records and (2) automatically measuring QT interval dispersion from the digitized records has been developed and validated. Hand measurements of the RR interval from the original tracing and cursor or automated measurements from digitized waveforms correlated to within 1%. QT intervals measured by cursor on digitized waveforms were a mean of 14 ms (95% confidence interval, 10-19 ms) longer than manual measurements on original tracings. Automatic QT interval measurements were a mean of 5 ms longer than cursor measurements (95% confidence interval, 3-7 ms). Automated measurements were observer independent and repeatable (coefficient of variation for repeat measurements, 0.137% RR and 0.370% QT). Estimates of QT dispersion (expressed as coefficient of interlead QT variation) were made for 14 patients with documented recurrent ventricular and 15 control subjects. The median coefficient of interlead QT variation was 8.8% (range, 4.4-12.4%) for arrhythmia patients and 3.6% (range, 2.7-6.3%) for the control group (P < .001). The automatic measurements were more conservative and less likely to give spuriously large values for QT dispersion than manual measurements. Automated QT dispersion measurements should facilitate future studies on predicting the risk of ventricular arrhythmias. PMID- 8228722 TI - Correlation between QTc interval duration and left ventricular systolic dysfunction in patients with acute myocardial infarction. AB - QTc interval and left ventricular regional wall motion abnormality score (WMS) were assessed in 61 consecutive patients with 4-day-old acute myocardial infarction (AMI). There were 45 men and 16 women, aged 60.6 +/- 8.7 years; 24 had anterior wall AMI and 37 had inferior wall AMI. Twenty-nine patients received thrombolytic treatment, which was presumably successful in 19. The longest QTc interval from a standard 12-lead electrocardiogram, corrected by Bazett's formula, was considered for the study. The WMS was evaluated echocardiographically. The authors found a statistically significant correlation between WMS and QTc interval duration; the correlation was stronger in patients with anterior AMI (r = .9, P < .001) than in those with inferior AMI (r = .66, P < .001). The WMS and QTc values were significantly lower in patients with successful thrombolytic treatment compared to those with unsuccessful thrombolysis (P < .003 and P < .002, respectively). The authors could demonstrate no significant correlation between serum potassium concentration and the QTc interval. In patients with anterior AMI, QTc interval duration might represent an additional marker of left ventricular systolic dysfunction. It could also be used as an additional noninvasive criterion of coronary artery reperfusion in patients with AMI. PMID- 8228723 TI - Intraatrial conduction delay associated with an epicardial implantable cardioverter-defibrillator. AB - The implantable cardioverter-defibrillator (ICD) has become an important mode of therapy for patients at risk for sustained ventricular arrhythmias. While the survival benefit of these devices is clear, adverse consequences are coming to light. The authors report a case of dramatic intraatrial conduction delay associated with repeated ICD shocks, resulting in pacemaker syndrome in a patient who had received both an ICD and a dual-chamber pacemaker. This and other conduction disturbances may become more common as experience with ICDs grows, and may demand adjunctive or alternative therapies to prevent frequent shocks. PMID- 8228724 TI - Theoretic effect of histamine on myocardium. PMID- 8228725 TI - Placental radar. PMID- 8228726 TI - Human recombinant inhibin A and testosterone act directly at the pituitary to suppress plasma concentrations of FSH in castrated rams. AB - The roles of inhibin and testosterone in the negative feedback control of the secretion of FSH were explored in experiments using castrated rams administered human recombinant inhibin A (hr-inhibin) and testosterone propionate (TP). Two experiments were conducted in the non-breeding season. In experiment 1, two groups of long-term castrated rams (wethers) were treated with an i.v. injection of either vehicle or hr-inhibin in two doses (25 and 50 micrograms) given 2 weeks apart. Plasma concentrations of FSH, measured by radioimmunoassay, were suppressed significantly (P < 0.01) and equally by both doses of hr-inhibin with a mean (+/- S.E.M.) maximal suppression of FSH of 19.9 +/- 2.60% occurring 6-10 h after injection. In experiment 2, hypothalamo-pituitary disconnected (HPD) wethers given 125 ng gonadotrophin-releasing hormone (GnRH) every 2 h, were treated with vehicle or 25 or 50 micrograms hr-inhibin before or after treatment (32 mg/day) with TP. A cross-over design was used so that each wether was treated with vehicle and hr-inhibin. Treatment with TP significantly (P < 0.001) suppressed plasma concentrations of FSH by 56%. Both doses of hr-inhibin were similarly effective in significantly (P < 0.05) suppressing plasma concentrations of FSH causing a mean suppression of 31.1 +/- 5.60% 6-10 h after injection. The suppressive effect of hr-inhibin was significantly (P < 0.05) increased when the wethers were treated with TP to a mean suppression of 50.7 +/- 5.6% 6-10 h after injection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228727 TI - Induction of the gonadotrophin surge-inhibiting factor by FSH and its elimination: a sex difference in the efficacy of the priming effect of gonadotrophin-releasing hormone on the rat pituitary gland. AB - This study was designed to explore the efficacy of gonadotrophin-releasing hormone (GnRH) to antagonize the effect of gonadotrophin surge-inhibiting factor (GnSIF) on the timing of the induction by GnRH of the maximal self-priming effect on pituitary LH responsiveness. The GnSIF levels were increased by FSH treatment and reduced after gonadectomy. Female rats were injected s.c. with 10 IU FSH or saline (control) on three occasions during the 4-day cycle. Serial i.v. injections of GnRH (500 pmol/kg body weight) were administered to intact rats on the afternoon of pro-oestrus or 15-30 min after ovariectomy. Intact male rats were given 10 IU FSH and 500 or 2000 pmol GnRH/kg body weight on an equivalent time-schedule. Endogenous GnRH release was suppressed with phenobarbital. In intact female control rats, the timing of the maximally primed LH response was delayed as the GnRH pulse-interval increased. FSH treatment of female rats induced a suppression of the initial unprimed LH response and delayed the maximally primed LH response, which showed further delay as the GnRH pulse interval was increased. When the pulsatile administration of GnRH was started 15 30 min after ovariectomy, the priming effect of GnRH did not change as the GnRH pulse-interval was increased in the saline-treated rats. However, FSH treatment caused a suppression of the unprimed LH response, a delay in the primed LH response and decreased the delay of the maximally primed LH response to GnRH when the GnRH pulse-interval was decreased. Increasing the interval between ovariectomy and the first GnRH pulse to 4 h diminished the efficacy of the FSH treatment: GnRH-induced priming was delayed by only one pulse instead of the two pulses in control rats. In intact males but not in orchidectomized rats, a self priming effect was demonstrated during GnRH pulses which were 1 h apart. The effect of 2 nmol GnRH/kg body weight was the most pronounced. Compared with intact female rats, the timing of the maximally primed LH response was delayed by 1 h. FSH treatment did not affect the pituitary LH response to both dose levels of GnRH. It is concluded that FSH treatment increased the release of GnSIF by the ovary, then induced a state of low responsiveness of the pituitary gland to GnRH and subsequently delayed GnRH-induced maximal self-priming. The efficacy of GnRH to prime the pituitary gland was higher when GnSIF levels were decreasing after removal of the ovaries.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8228728 TI - Antiuterotrophic effects of the pure antioestrogen ICI 182,780 in adult female monkeys (Macaca nemestrina): quantitative magnetic resonance imaging. AB - The antiuterotrophic efficacy of the pure antioestrogen ICI 182,780 has been demonstrated previously by magnetic resonance imaging (MRI) in ovariectomized oestrogen-treated monkeys (Macaca nemestrina). Further characterization of the effects of ICI 182,780 in intact adult female monkeys with normal menstrual cycles was undertaken to provide an indication of its potential actions in premenopausal women. Changes in the volume of uterine tissues were measured by MRI in early, mid and late cycle. The volume of the uterus varied up to fivefold between individual monkeys but serial observations in individuals provided sufficient precision to allow accurate assessments to be made of changes in the endometrium and myometrium during the course of the menstrual cycle and following ICI 182,780 administration. In comparison with its initial size in untreated monkeys, the endometrium increased in volume by 60% and 125% in the mid and late cycle respectively. In contrast, the size of the myometrium decreased significantly, by 16% from early to mid cycle and then recovered to near its initial volume in the late cycle. Treatment with ICI 182,780 beginning in the early part of the menstrual cycle prevented the growth of the uterus. The magnitude and duration of the response was dependent on whether or not ovulation occurred during treatment with ICI 182,780. In animals rendered anovulatory, growth of the endometrium was blocked completely by ICI 182,780 and the volume of the tissue declined below that present at the start of the menstrual cycle. Antiuterotrophic efficacy was significantly less in monkeys which ovulated during treatment with ICI 182,780.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228729 TI - Influence of endogenous cholinergic tone and alpha-adrenergic pathways on growth hormone responses to His-D-Trp-Ala-Trp-D-Phe-Lys-NH2 in the dog. AB - His-D-Trp-Ala-Trp-D-Phe-Lys-NH2 (GHRP-6) is a synthetic peptide unrelated to any known hypothalamic-releasing hormone including growth hormone-releasing hormone (GHRH). Interestingly, this peptide induces a dose-related increase in plasma GH levels in all species tested so far. The aim of this study was to investigate the action of GHRP-6 alone or in combination with GHRH on GH release in dogs. In addition, the activation or blockade of endogenous cholinergic tone and alpha-1 adrenoceptors on GHRP-6-stimulated GH secretion was assessed. In adult Beagle dogs (n = 10), GHRP-6 (90 micrograms i.v.) increased basal GH levels from 2.6 +/- 1.5 to 14.4 +/- 3.1 micrograms/l (mean +/- S.E.M.) after 15 min. GHRH (50 micrograms i.v.) induced a GH peak of 9.7 +/- 2.2 micrograms/l at 15 min. The combined administration of GHRP-6 and GHRH strikingly potentiated canine GH release with a peak of 54 +/- 9.0 micrograms/l (P < 0.01). Pretreatment with the cholinergic agonist pyridostigmine (30 mg per os) increased GHRP-6-stimulated GH secretion (37.9 +/- 10.1 micrograms/l P < 0.05), while the muscarinic blocker atropine (100 micrograms i.v.) completely abolished (GH peak lower than 2 micrograms/l) the stimulatory action of GHRP-6. On the other hand, administration of the alpha-2 adrenergic agonist clonidine (4 micrograms/kg i.v.) increased basal plasma GH levels without affecting GH responses to GHRP-6.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228730 TI - Changes induced by cadmium administration on thyroxine deiodination and sulfhydryl groups in rat liver. AB - The effects of cadmium on 5'-deiodination of thyroxine (T4) by rat liver and on the hepatic concentration of non-protein sulfhydryl groups (NPSH) were studied in Wistar rats of 200-250 g body weight. A group of ten rats was injected with cadmium chloride (300 micrograms/100 g body weight i.p.) daily for 4 days. Another group of six rats received, in addition, dithiothreitol (DTT; 1 mg/100 g body weight i.p.) daily for the same period. A group of eight normal untreated rats served as control. T4 deiodination was also determined in aliquots of liver from untreated rats, with cadmium (2 or 5 mmol/l) and with or without DTT (0, 2.5, 5 or 10 mmol/l) plus 1 microCi 125I-labelled T4. Hepatic NPSH were measured by a colorimetric method employing dithioldinitrobenzoic acid. Homogenates were incubated for 90 min at 37 degrees C and chromatographed in a tertiary amyl alcohol:hexane:ammonia (2 mol/l) (10:1:12) system. Cadmium-injected rats showed a significant (P < 0.01) decrease in T4 deiodination and in the generation of 125I (P < 0.01) and tri-iodothyronine (T3) (P < 0.02). NPSH were also decreased (P < 0.02). Administration of DTT restored T4 deiodination and NPSH to normal. In vitro addition of cadmium or DTT to normal rat liver homogenates induced similar effects on the degradation of T4. Serum concentrations of T4 (P < 0.01) and T3 (P < 0.01) declined significantly in cadmium-injected rats, whereas DTT administration failed to normalize serum hormone levels. The data suggest that cadmium may have decreased 5'-deiodinating activity through binding to sulfhydryl groups of 5'-deiodinase as it does in other enzymes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228731 TI - Thyrotrophin-releasing hormone-induced growth hormone secretion in domestic fowl: concomitant stimulation of dopamine turnover in the medial basal hypothalamus. AB - Thyrotrophin-releasing hormone (TRH) stimulates GH secretion in domestic fowl by actions at pituitary and central nervous system sites. The possibility that this central action might be mediated by hypothalamic catecholamines or indoleamines was therefore investigated. When TRH was administered into the lateral ventricles of anaesthetized fowl the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC, a metabolite of dopamine (DA)) in the medial basal hypothalamus (MBH) was increased within 20 min. The concentrations of MBH noradrenaline (NA), DA, serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were, however, unaffected by the intra-cerebroventricular (i.c.v.) administration of TRH, although the MBH concentrations of somatostatin and TRH were concomitantly reduced. A rapid increase in DA release into MBH extracellular fluid and its metabolism to DOPAC was also observed after i.c.v. or i.v. administration of TRH, in birds in which the MBH was perfused in vivo with Ringer's solution. Microdialysate concentrations of NA, 5-HT and 5-HIAA were not, however, affected by central or peripheral injections of TRH. Diminished GH responses to i.v. TRH challenge occurred in birds pretreated with reserpine (a catecholamine depletor), alpha-methyl-paratyrosine (a DA synthesis inhibitor) and pimozide (a DA receptor antagonist). These results therefore provide evidence for the involvement of a hypothalamic dopaminergic pathway in the induction of GH release following the central or peripheral administration of TRH. In contrast with its inhibitory actions at peripheral sites, DA would appear to have a central stimulatory role in regulating GH release in birds. PMID- 8228732 TI - Receptors for insulin-like growth factor-II in the growing tip of the deer antler. AB - Insulin-like growth factor-II (IGF-II) binding in the growing tip of the deer antler was examined using autoradiographical studies, radioreceptor assays and affinity cross-linking studies. Antler tips from red deer stags were removed 60 days after the commencement of growth, and cryogenically cut into sections. Sections were incubated with radiolabelled IGF-II, with or without an excess of competing unlabelled IGF-II and analysed autoradiographically. Radiolabelled IGF II showed high specific binding in the reserve mesenchyme and perichondrium zones, which are tissues undergoing rapid differentiation and cell division in the antler. Binding to all other structural zones was low and significantly (P < 0.001) less than binding to the reserve mesenchyme/perichondrium zones. Radioreceptor assays on antler microsomal membrane preparations revealed that the IGF-II binding was to a relatively homogeneous receptor population (Kd = 1.3 x 10(-10) mol/l) with characteristics that were not entirely consistent with those normally attributed to the type 2 IGF receptor. Tracer binding was partly displaceable by IGF-I and insulin at concentrations above 10 nmol/l. However, affinity cross-linking studies revealed a single band migrating at 220 kDa under non-reducing conditions, indicative of the type 2 IGF receptor. These results indicate that, in antler tip tissues, IGF-II binds to sites which have different binding patterns and properties from receptors binding IGF-I. This may have functional significance as it appears that, whilst IGF-I has a role in matrix development of cartilage, IGF-II may have a role in the most rapidly differentiating and proliferating tissues of the antler. PMID- 8228733 TI - Bioactive and immunoreactive variants of prolactin in milk and serum of lactating rats and their pups. AB - Prolactin (PRL)-like bioactivity (in Nb2 lymphoma assay) and immunoreactivity (in radioimmunoassay (RIA)) in rat milk, maternal and neonatal sera and in neonatal rat pituitary cultures were investigated. The PRL-like bioactivity in the water soluble fraction of rat milk was high and exceeded its immunoreactivity 5.8-, 4.0 and 2.1-fold, on days 2, 12 and 22 of lactation respectively. The elevated bioactivity to immunoreactivity (B/I) ratio of PRL in milk was not due to the presence of interleukin-2 (IL-2) in milk, since the proliferation of the CTLL-2 murine T cells, which are not sensitive to PRL, was promoted by IL-2 but not by milk. Serum levels of immunoreactive PRL were low in sera of non-weaned rat pups on days 2, 12 and 22 postpartum. Similar to milk, the B/I ratio of PRL in sera of rat pups was high and decreased with time postpartum. Pituitary glands of pups obtained on days 2, 12 and 22 secreted progressively increasing amounts of PRL in vitro; the B/I ratio ranged between 1.2 and 2.1 without a significant change. The relative concentrations of size variants in milk were not proportional to those in serum of lactating rats on day 2 postpartum as assessed by Sepharcryl S-100 HR gel permeation chromatography and Nb2 bioassay or RIA. Size variants of biologically active PRL were abundant in early milk and gradually diminished as lactation progressed: a partially resolved peak representing monomeric to dimeric PRL variants (relative molecular weights ranging between 18 k and 42 k) became progressively narrower between days 2 and 22. Biologically active and immunoreactive PRLs displayed disparate elution profiles. The elution profile of PRL in sera of neonatal rats on day 2 post-partum was different from that of maternal serum or milk. The major immunological (and possibly biological) PRL like activity eluted as two adjacent peaks at 2.2 k and 1.5 k, raising the possibility that fragments of milk-borne PRL were absorbed from the gut after partial proteolytic degradation. In contrast with PRL, GH (which is present in rat milk only in minute concentrations) did not show heterogeneity in sera of 2 day-old rat pups in gel permeation chromatography. The present results demonstrate that the concentrations of PRL-like activity in rat milk and newborn rat serum have been grossly underestimated because levels have been measured by RIA. The high B/I ratio of PRL in milk and neonatal sera is due to the presence of PRL-related compounds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228734 TI - Characterization of a novel non-peptide vasopressin V1 receptor antagonist (OPC 21268) in the rat. AB - A non-peptide, orally effective, vasopressin (AVP) V1 receptor antagonist 1-(1-[4 (3-acetylaminopropoxy) benzoyl]-4-piperidyl)-3,4-dihydro-2(1H)-quinolinone (OPC 21268) has recently been described. This paper reports the in-vitro and in-vivo characterization of OPC-21268 binding to vasopressin receptors in rat liver and kidney. OPC-21268 caused a concentration-dependent displacement of the selective V1 receptor antagonist radioligand, 125I-labelled [d(CH2)5,sarcosine7]AVP to V1 receptors in both rat liver and kidney medulla membranes. The concentration of OPC-21268 that displaced 50% of specific AVP binding (IC50) was 40 +/- 3 nmol/l for liver V1 and 15 +/- 2 nmol/l for kidney V1 receptors (mean +/- S.E.M.; n = 3). OPC-21268 had little effect on the selective V2 antagonist radioligand [3H]desGly-NH2(9)]d(CH2)5,D-Ile2,Ile4] AVP binding to V2 receptors in renal medulla membranes (IC50 > 0.1 mmol/l). After oral administration to rats, OPC 21268 was an effective V1 antagonist in a time- and dose-dependent manner. Binding kinetic studies showed that OPC-21268 acted as a competitive antagonist at the liver V1 receptor in vitro and in vivo, in addition to its in-vitro competitive effects at the renal V1 receptor. OPC-21268 shows promise as an orally active V1 antagonist. PMID- 8228735 TI - Growth hormone receptor regulation in growth hormone-deficient dwarf rats. AB - In the rat, many actions of GH depend upon the sexually dimorphic pattern of exposure to GH. Hepatic human GH (hGH) receptor binding differs between the sexes and is sensitive to GH deficiency, but this has mostly been studied in acutely hypophysectomized rats, which lack all pituitary hormones. We have used a strain of GH-deficient dwarf (Dw) rats to determine whether chronic GH deficiency alters the normal developmental pattern and sexually dimorphic expression of hepatic GH receptors. Adult female Dw rats had lower levels of 125I-labelled hGH binding (reflecting predominantly lactogenic receptors) than their normal counterparts whereas there was no difference between adult Dw and normal males; binding capacity increased from 25 days of age, becoming sexually dimorphic from 40 days to adulthood in both strains (% specific binding/mg protein: normal males 1.6 +/- 0.3, normal females 13.2 +/- 1.1, Dw males 2.1 +/- 0.4, Dw females 10.0 +/- 0.6). In contrast, hepatic 125I-labelled bovine GH (bGH) binding (somatogenic receptors) was much lower, and similar in both Dw and normal animals. A sex difference in 125I-labelled bGH binding was only seen in adult animals, and was considerably less marked in Dw rats compared with normal animals (normal males 1.3 +/- 0.1, normal females 2.5 +/- 0.2, Dw males 1.9 +/- 0.2, Dw females 2.4 +/- 0.2%/mg protein). Continuous hGH infusion stimulated growth in female Dw rats, and raised somatogenic and lactogenic GH binding (3.2 +/- 0.4 and 19.6 +/- 2.5%/mg protein) compared with sham-infused controls (2.4 +/- 0.2 and 7.9 +/- 0.6%/mg protein).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228736 TI - The effect of dietary protein restriction on the secretory dynamics of 1 alpha hydroxycorticosterone and urea in the dogfish, Scyliorhinus canicula: a possible role for 1 alpha-hydroxycorticosterone in sodium retention. AB - The putative osmoregulatory role of the unique elasmobranch corticosteroid, 1 alpha-hydroxycorticosterone (1 alpha-OH-B), was investigated using dietary protein restriction as a means of limiting urea biosynthetic ability. Groups of dogfish (Scyliorhinus canicula) were adapted to either a high or a low protein diet (HPD and LPD respectively) and the secretory dynamics of urea and 1 alpha-OH B were determined following acclimation to normal (100%), 130% and 50% sea water. In normal sea water, LPD fish showed significantly decreased blood production of urea compared with fish fed a HPD (P < 0.05), and the plasma urea concentration required to maintain iso-osmolality was achieved only by a substantial decrease in urea clearance from the plasma. Unlike HPD fish, LPD fish in 130% sea water had no apparent ability to increase plasma urea concentration. An alternative strategy adopted by these animals was the retention of high plasma concentrations of Na+ and Cl-, which increased plasma osmolality and tended to decrease osmotic water loss. Concomitant with the increased ion concentrations, plasma 1 alpha-OH B concentration was also greatly elevated in LPD fish indicating that the steroid may be acting to minimize Na+ (and Cl-) excretion at osmoregulatory sites such as the rectal gland, kidney and gills. This and a previous study have also demonstrated that 1 alpha-OH-B concentration is elevated in 50% sea water. Decreases in plasma Na+ concentration are tolerated down to 75% sea water, whereafter Na+ is preferentially retained and further decreases in osmolality are achieved by reductions in plasma urea concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228737 TI - Distinct regional localization of neurocalcin, a Ca(2+)-binding protein, in the bovine adrenal gland. AB - Neurocalcin (molecular weight 23,000 and 24,000) is a Ca(2+)-binding protein with three putative Ca(2+)-binding domains and is present in large amounts in nervous tissues. Neurocalcin isoproteins separated by C18 reverse-phase column chromatography are insoluble in buffer solution and it is impossible to determine the dissociation constant of neurocalcin with Ca2+. To overcome this difficulty, recombinant neurocalcin was synthesized, based on one of the cDNAs of the neurocalcin isoproteins. Stoichiometric titration experiments, using recombinant neurocalcin, indicated that this protein bound 2 mol Ca2+/mol protein and that the apparent dissociation constant for Ca2+ was 2.2 mumol/l, suggesting that neurocalcin plays a physiological role in cellular function. Immunoblotting showed that neurocalcin is present in the bovine adrenal gland in addition to the nervous tissues. Neurocalcin, identified by immunoblotting, was purified from the bovine adrenal gland. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of neurocalcin from the bovine brain showed 23 kDa and 24 kDa double bands, while SDS-PAGE of neurocalcin from the adrenal gland showed a single band of apparently 24 kDa, suggesting that the expression of neurocalcin isoproteins differs from tissue to tissue. The content of neurocalcin in the adrenal gland was 10 micrograms protein/100 g wet tissue. Immunohistochemical analysis showed the occurrence of neurocalcin in zona glomerulosa and adrenal medulla but not in zona fasciculata or zona reticularis. The restricted localization of neurocalcin in the adrenal gland suggests that a similar Ca2+signal pathway may be present in zona glomerulosa a nd the adrenal medulla. PMID- 8228738 TI - The luteotrophic actions of prostaglandins E2 and F2 alpha on dispersed marmoset luteal cells are differentially mediated via cyclic AMP and protein kinase C. AB - Progesterone production by dispersed luteal cells obtained from the marmoset monkey on day 14 after ovulation can be stimulated by both prostaglandin F2 alpha (PGF2 alpha) and its structural analogue, cloprostenol. To establish whether these responses can be attributed to cross-reaction with the prostaglandin E2 (PGE2) receptor, this study compared the involvement of cyclic adenosine-3',5' monophosphate (cAMP) and protein kinase C (PKC) in the luteotrophic responses to PGE2, PGF2 alpha and cloprostenol. While all three prostaglandins stimulated similar increases in progesterone production (239.5 +/- 7.9% of control; P < 0.01), only PGE2 stimulated a significant increase in cAMP accumulation (373.2 +/ 28.4% of control; P < 0.01). This study is the first to demonstrate PKC activity in the marmoset ovary. Following down-regulation of PKC with a tumour-promoting phorbol ester, 4 beta-phorbol 12-myristate 13-acetate (4 beta-PMA), basal progesterone production was significantly increased (150.9 +/- 8.2% of control; P < 0.05) and the luteotrophic effects of PGF2 alpha and cloprostenol were no longer evident, whereas the response to PGE2 was unaffected. These observations are consistent with the differential involvement of cAMP and PKC in the luteotrophic responses to PGE2 and PGF2 alpha/cloprostenol respectively. Hence, we conclude that the luteotrophic actions of prostaglandins E2 and F2 alpha on dispersed marmoset luteal cells are mediated via different receptors and signal transduction pathways. PMID- 8228739 TI - Expression of mRNA coding for pituitary hormones and pituitary-specific transcription factor in the pituitary gland of the rdw rat with hereditary dwarfism. AB - The rdw rat (gene symbol: rdw) with hereditary dwarfism has been shown immunohistochemically to have subnormal numbers not only of GH- but also of prolactin- and thyrotrophin-positive cells. To characterize the dwarfism of this strain, the expression of pituitary hormone mRNAs was examined by Northern hybridization. The pituitary gland in the rdw rat expressed 30-100 times less GH and prolactin mRNAs than normal controls, whereas mRNAs for pro-opiomelanocortin and the alpha subunit of rat glycoprotein hormone revealed a significant increase. There was a non-significant difference in rat LH-beta subunit and FSH beta subunit between normal and rdw rats. The suppressed expression of a pituitary-specific transcription factor, Pit-1, is considered to cause hereditary dwarfism in mouse strains Snell and Jackson, whose phenotypes resemble those of the rdw rat. In this study, however, no difference in mRNA expression for Pit-1 was found between rdw rats and controls. This work indicates that the rdw rat may not have the same genotype as the phenotypically similar dwarf mice, Snell, Jackson and Ames. PMID- 8228740 TI - Effect of chronic FSH administration on ovarian follicular development, ovulation rate and corpora lutea formation in sheep. AB - This study in ewes examined the effects on ovarian function of a pulsatile regimen of ovine FSH (NIADDK-oFSH-17) administered over a 24- to 28-day period beginning on day 1 of the oestrous cycle (day 0 = oestrus). The FSH (1.66 micrograms or 5.00 micrograms) was administered i.v. over a 1-min interval once every hour throughout the treatment period. In other ewes ovine LH (NIDDK-oLH-23) was administered (10 micrograms once every 2 h) for 24-28 days together with oFSH (1.66 micrograms/h). Compared with untreated controls (n = 19 ewes), FSH alone at both doses (n = 10 ewes/dose) as well as the FSH + LH treatment (n = 10) led to significant increases in the plasma concentrations of FSH (P < 0.01), ovarian weight (P < 0.05) and ovulation rate (P < 0.01) but there was no change in the mean weight of individual corpora lutea (CL). Exogenous FSH at the high but not the low dose alone or with LH stimulated a significant overall increase in plasma inhibin concentrations (P < 0.05). The geometric mean (and 95% confidence limits) ovulation rates in the high FSH (i.e. 5.00 micrograms/h), low FSH (i.e. 1.66 micrograms/h), low FSH + LH, and control treatment groups were 15.3 (9.3, 24.8), 3.7 (2.1, 6.0), 3.7 (2.5, 5.8) and 1.4 (1.2, 1.7) respectively. The FSH or FSH + LH treatments did not alter the total numbers of antral follicles (> or = 1 mm diameter). However, the high but not the low FSH or low FSH + LH treatment led to significant increases in the mean numbers of large follicles (i.e. > 4.5 mm diameter; P < 0.01) and a higher proportion of non-atretic antral follicles. Highly significant linear relationships were found between the mean plasma concentrations of FSH or inhibin and the ovulation rate (FSH: r = 0.74, P < 0.0001; inhibin: r = 0.93, P < 0.0001). Highly significant linear relationships were also found between the plasma concentrations of FSH or inhibin and the number of large follicles (i.e. > 4.5 mm diameter; FSH, r = 0.78, P < 0.0001; inhibin, r = 0.80, P < 0.0001) and between the plasma concentrations of inhibin and the number of granulosa cells in large follicles (r = 0.78, P < 0.0001). After the high FSH but not the low FSH treatment there were significant increases in both FSH- and LH-induced responsiveness in granulosa cells with respect to cyclic AMP synthesis in vitro.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8228741 TI - Variability in the urinary excretion of growth hormone in children: a comparison with other urinary proteins. AB - As a basis for assessment of the clinical validity of urinary GH (uGH) measurements in children, the day-to-day variability in renal handling of GH has been compared with that of albumin, N-acetylglucosaminidase (NAG) and creatinine. Five overnight urine specimens were collected over a 2-week period from 78 healthy children (aged 5-16 years), 20 of normal stature and 58 with growth disorders; ten children were classified as GH-deficient (GHD) and 48 were designated short normal (SN). The variability of excretion of each substance was expressed as a coefficient of variation (C.V.) which was not influenced by expressing the urine results as total mass excreted, concentration, excretion rate or as a ratio to creatinine. There was considerable night-to-night variability in the excretion of all substances (mean C.V. values for all groups: 56% for albumin, 41% for GH, 33% for NAG and 27% for creatinine). No differences were found in the variability of GH excretion between males and females, nor between prepubertal and pubertal subjects. The mean C.V. for uGH excretion ranged from 37% in normal and 35% in SN children to 52% in those with GHD (P < 0.05). Assay variation rather than a change in renal protein handling accounted for the large variations in uGH concentrations of < 5 pg/ml, thus contributing to the high uGH C.V. of the GHD group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228742 TI - The Second International Standard for Human Pituitary LH: its collaborative study by bioassays and immunoassays. AB - The second International Standard for Human Pituitary LH (in ampoules coded 80/552; 2nd IS) and LH 81/535 (prepared in the same way as the 2nd IS from the same LH preparation) were compared with the International Reference Preparation of Human Pituitary LH for Immunoassay (IRP 68/40) by 19 laboratories in 11 countries, using in-vivo and in-vitro bioassays, a receptor assay and immunoassays. Geometric mean estimates of the LH content of the 2nd IS (with 95% fiducial limits) in terms of IRP 68/40 were: 34.6 (29.1-41.0) IU/ampoule by in vivo bioassays; 35.8 (27.0-47.4) IU/ampoule by in-vitro bioassays; 58.6 IU/ampoule by one receptor assay; and 36.8 (35.5-38.1) IU/ampoule by immunoassays. The close agreement between the relative activities of the 2nd IS and IRP 68/40 in the wide range of assay systems studied appears to reflect the fact that both standards contain highly purified LH with similar isoform compositions as judged by isoelectric focusing. Estimates of the LH content of LH 81/535 in terms of IRP 68/40 and in terms of the 2nd IS tended to be lower than those for the 2nd IS across all methods, but the differences were not statistically significant. The 2nd IS was found to be as suitable as IRP 68/40 as a standard for the in-vitro bioassay and immunoassay of LH in the two serum samples studied. However, the mean estimates of serum LH in terms of either of these standards were more than 150% larger by in-vitro bioassays than by immunoassays and more than 50% larger by one-site than by two-site immunoassays.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228743 TI - Adaptation of thermafil components to canal walls. PMID- 8228744 TI - Sealing of human dentinal tubules by XeCl 308-nm excimer laser. AB - Root hypersensitivity occurs as a result of exposed dentinal tubules. Various methods and materials have been tried in an attempt to occlude these tubules. The purpose of this investigation was to study by scanning electron microscope the effects of XeCl excimer laser on exposed dentinal tubules of human extracted teeth. Fifteen 3-mm-thick slices were cut at the cementoenamel junction from 15 extracted human teeth by an electric saw. By using a diamond bur to remove the cementum layer the dentinal tubules were exposed. Each slice was scored by a permanent marker into four equal quadrants. Three of the quadrants were lased for 4 s by XeCl excimer laser with fluences ranging from 0.5 to 7.0 J/cm2 and pulse repetition of 25 Hz. The unlased quadrant served as control. The specimens were mounted on a stub, sputter coated by gold, and examined by scanning electron microscope. Nonlased surfaces showed numerous exposed dentinal tubules. In contrast, all specimens lased at fluences of up to 1 J/cm2 showed the presence of melted dentin which closed the dentinal tubules. At fluences of 4 J/cm2 and higher, rupture of molten materials and exposure of dentinal tubules were noted. The results indicate the application of XeCl excimer laser at specific fluences can cause melting of dentin and closure of exposed dentinal tubules. PMID- 8228745 TI - Preliminary report on the application of pulsed CO2 laser radiation on root canals with AgCl fibers: a scanning and transmission electron microscopic study. AB - Fifteen maxillary central incisors were treated in vitro with pulsed CO2 laser radiation (wavelength:9.6-microns pulse duration:135-microseconds pulse energy:60 mJ energy density:12 J/cm2) delivered by an AgCl fiber into the root canal. Preliminary results show opening of dentin tubules as well as fused areas of hydroxyapatite in the root canal after laser treatment. Temperature measurement at the root surface showed that 40 degrees C was not exceeded. These preliminary results show the ability of this laser system to open dentin tubules and to fuse hydroxyapatite but further development in fiber technology is necessary to achieve predictable results. PMID- 8228746 TI - Effect of moisture on the apical seal of laterally condensed gutta-percha. AB - Eighty-five single-rooted human teeth were cleansed and shaped for obturation with gutta-percha and Proco-Sol root canal cement. Two of the three experimental groups had their canals contaminated with either 0.05 ml of NaOCl or saliva immediately prior to obturation. The remaining group of 25 teeth had dry canals prior to obturation. All teeth were obturated by the lateral condensation technique, covered with two coats of nail polish, immersed in India ink, and subjected to a vacuum for 15 min. The teeth were then cleared to allow visual assessment of the dye penetration through the apical seal. Positive and negative controls verified the experimental method. The recorded measurements were statistically analyzed and no significant differences were found among the groups. PMID- 8228747 TI - Comparison by means of a computer-supported device of the enlarging characteristics of two different instruments. AB - Numerous studies report different results when testing endodontic instruments with similar and/or different research protocols. However, objective studies that could eliminate the factors inherent to the operator have not yet been developed. Simulated root canals in clear casting resin were used as models. The Canal Master U and Flexogate were tested by using a double exposure photographic technique. A computer-aided device capable of reproducing three-dimensional enlarging movements and irrigation of the simulated canals was developed. The enlarging movements were programmed according to the step-back and step-down techniques. The enlarging of simulated root canals with this device proved to be constant and reproducible. Standard deviations were kept low and were dependent on the degree of canal curvature. The enlarged simulated root canals showed that both instruments could be kept centered in the path of the root canal. Results showed only a few significant differences between the effective material removed and a theoretical ideal amount of removed material. PMID- 8228748 TI - Cytotoxicity of retrofill materials. AB - Dentin bonding agents reduce microleakage and enhance marginal adaption of composite resin restorations. These characteristics are advantages for their use as an endodontic retrofilling material. Because these materials will be in direct contact with vital tissues, their cytotoxic potential must be evaluated before clinical use. It was the purpose of this study to evaluate the cell cytotoxicity of amalgam, Caulk Universal Bond, Gluma, 35% HEMA, Morita Clearfil, Scotchbond 2, Super EBA, Tenure, and Tenure 5-4. VERO cells were grown in RPMI-1640 medium and cell monolayers were prepared by incubating 15 ml of the cell suspension in 60-mm culture dishes at 37 degrees C in 5% CO2. Twelve milliliters of a medium-agarose mixture containing 1% neutral red vital stain were overlayed onto the cell layer and allowed to solidify. The materials were directly exposed to the agarose overlays by inverting 6.0-mm diameter polypropylene capsules containing the cured and liquid sample materials either immediately (0 time) or after placement in phosphate-buffered saline with 1% gentamicin for 7, 15, or 30 days. Cytotoxicity was determined by measuring the zone of killed cells around the sample 24 h after placement on the agarose. Cytotoxicity was determined by measuring the zones of cell inhibition at 24 h and at 7, 15, and 30 days. Initially, all of the materials were found to be cytotoxic, except amalgam and the Tenure components. The dentin bonding primers showed a mean zone of inhibition of 13.2 mm and the cleansers a 40.0-mm zone. Amalgam demonstrated increasing cytotoxicity: 0.0 mm at 24 h to 12.0 mm at 30 days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228749 TI - Dye ingress in molars: furcation to chamber floor. AB - An in vitro dye leakage study was performed to determine the incidence of patent furcal accessory canals following exposure of the pulp chamber to 5.25% sodium hypochlorite. One hundred extracted molars were labeled, endodontically opened, and irrigated for 1 h at 5-min intervals. The external furcations were exposed to 0.5% basic fuchsin for 24 h. Patency was determined by dye presence on the chamber floor. Statistical analysis revealed that first and second molars, regardless of arch position or location, demonstrated a patent accessory canal at a rate of 57% +/- 19.6% in the furcal area. It was concluded that accessory canal exposure to 5.25% sodium hypochlorite in the furcal area of molars produced patency which was demonstrable via passive methods--no vacuum or injection pressures were utilized. PMID- 8228750 TI - Migration of a Streptococcus sanguis strain through the root dentinal tubules. AB - The persistent presence of bacteria in the root canal system often leads to the failure of treatment. The aim of this study was to study the in vitro penetration of root dentinal tubules by bacteria. Bovine dentin was inoculated with a strain of Streptococcus sanguis, NCTC 7863; samples were taken at 10, 20, and 28 days, and bacterial penetration of the tubules was demonstrated using two techniques, scanning electron microscopy and light microscopy after Brown and Brenn staining. The results showed that S. sanguis can penetrate deep inside the tubules (792 microns). The maturity of the dentin blocks seems to be a major factor in tubule penetration. PMID- 8228751 TI - pH changes in root dentin over a 4-week period following root canal dressing with calcium hydroxide. AB - Root canals in extracted human teeth were cleaned and shaped and subsequently dressed with a calcium hydroxide root canal dressing. pH Changes in the root dentin were measured over a 4-wk period with microelectrodes in small cavities at apical and cervical levels in inner and outer dentin. The pH increased within hours in the inner dentin, peaking at pH 10.8 cervically and 9.7 apically. However, 1 to 7 days elapsed before the pH began to rise in the outer root dentin, reaching peak levels of pH 9.3 cervically and 9.0 apically after 2 to 3 wk. The results show that hydroxyl ions derived from a calcium hydroxide dressing do diffuse through root dentin. They diffuse faster and reach higher levels cervically than apically. Surface pH measurements showed that hydroxyl ions do not diffuse in more than a minor way through the intact root surface. PMID- 8228752 TI - Postsurgical endodontics. AB - Apical surgery is known to be an effective and practical way of treating endodontic failures. However, in spite of seemingly proper root end management, surgical failures may occur as a result of lateral canal, dentinal tubule, end retrograde leakage. Since surgical retreatment of such failures would likely result in failure, the alternative of a nonsurgical retreatment should be carefully considered. An understanding of the relationship which exists between the quality of the orthograde treatment and the surgical prognosis is necessary to properly direct the course of treatment. PMID- 8228753 TI - Reliability of electric pulp testing after pulpal testing with dichlorodifluoromethane. AB - The purpose of this study was to determine if the sequence and interval between electric pulp testing and cold vitality testing with dichlorodifluoromethane affects the reliability of pulpal diagnostic testing. Sixty vital teeth in 15 volunteers were tested. Ten endodontically treated teeth were used as negative controls. After isolation and asepsis techniques, baseline threshold responses from a digital electric pulp tester were recorded from the maxillary incisors. A dichlorodifluoromethane-saturated cotton pellet was applied to teeth 8, 9, and 10. Electric pulp testing was repeated at 30-s, 1-min, and 2-min intervals on all test teeth after the cold test. The level of responses were recorded and statistically analyzed. The results of this study indicate that electric pulp testing is not adversely affected by the use of dichlorodifluoromethane. PMID- 8228754 TI - Incidence of periradicular pathoses in endodontic treatment failures. AB - Biopsy reports from 150 periradicular tissue specimens obtained from teeth refractory to nonsurgical endodontic therapy were reviewed. The specimens were submitted by postdoctoral dental students in the Department of Endodontics, and the biopsy reports were prepared by oral pathologists at the University of Texas Health Science Center at San Antonio. The study found that 59.3% of the periradicular lesions were granulomas, 22% cysts, 12% scars, and 6.7% other pathoses. The majority (56%) of endodontically treated cases which failed to heal were recognized within 2 yr after the completion of therapy. The most common location for surgical retreatment was the anterior maxilla, followed by the posterior maxilla, the posterior mandible, and the anterior mandible. The periapical granuloma was the predominant pathosis at each location. PMID- 8228755 TI - Safe and easy way to use calcium hydroxide as a temporary dressing. PMID- 8228756 TI - Research and development in internal medicine in the United Kingdom. Interview by Stuart Donnan. PMID- 8228757 TI - Thinness at birth in a northern industrial town. AB - OBJECTIVE: To determine whether babies in an area of Britain with unusually high perinatal mortality have different patterns of fetal growth to those born elsewhere in the country. DESIGN: Measurement of body size in newborn babies. SETTING: Burnley (perinatal mortality in 1988 15.9/1000 total births) and Salisbury (perinatal mortality 10.8/1000 total births), England. SUBJECTS: Subjects comprised 1544 babies born in Burnley, Pendle, and Rossendale Health District, and 1025 babies born in Salisbury Health District. MAIN OUTCOME MEASURES: Birthweight, length, head, arm and abdominal circumferences, and placental weight were determined. RESULTS: Compared with babies born in Salisbury, Burnley babies had lower mean birthweight (difference 116 g, 95% confidence interval (CI) 77,154), smaller head circumferences (difference 0.3 cm, 95% CI 0.2, 0.4), and were thinner as measured by arm circumference (difference 0.3 cm, 95% CI 0.3, 0.4), abdominal circumference (difference 0.5 cm, 95% CI 0.4, 0.6) and ponderal index (difference 0.8 kg/m3, 95% CI 0.6, 1.0). The ratio of placental weight to birthweight was higher in Burnley (difference 0.6%, 95% CI 0.4, 0.9). These differences were found in boys and girls and did not depend on differences in duration of gestation or on the different ethnic mix of the two districts. Mothers in Burnley were younger, shorter in stature, had had more children, were of lower social class, and more of them smoked during pregnancy than mothers in Salisbury. These differences did not explain the greater thinness of their babies. CONCLUSIONS: Babies born in Burnley, an area with high perinatal mortality, are thin. The reason is unknown. Poor maternal nutrition is suspected because Burnley babies have a higher ratio of placental weight to birthweight. The greater thinness at birth of Burnley babies could have long term consequences, including higher rates of cardiovascular disease. PMID- 8228758 TI - Influence of socioeconomic and health care development on infant and perinatal mortality in Spain 1975-86. AB - STUDY OBJECTIVE: This study aimed to analyse the influence of social, economic, and health development on infant and perinatal mortality in Spain between 1975 and 1986, and to identify possible changes in these relationships over time. DESIGN: Study of the association between mortality and a range of variables. SETTING: 50 Spanish provinces. MEASUREMENTS AND MAIN RESULTS: Mean infant and perinatal mortality were estimated for two periods--1975-8 and 1983-6. Social, economic, and health care indicators were collected as independent variables for these two periods. The rates of variation between periods were estimated for each variable. Multiple linear regression models were used to define the association between infant and perinatal mortality and their respective rate of variation with the former indicators. Mean familial income was the main predictive factor for infant and perinatal mortality in the first period but in the second period health care indicators were more relevant. CONCLUSIONS: The reduction in Spanish infant and perinatal mortality over the period can be attributed mainly to the improvement in prenatal and neonatal health care in Spain in recent years, while economic factors seem less important. PMID- 8228759 TI - Video display terminal use during pregnancy and reproductive outcome--a meta analysis. AB - STUDY OBJECTIVE: The aim was to obtain quantitative information from published data on the potential association between video display terminal (VDT) use during pregnancy and the outcome. DESIGN: Results of nine published case-control studies (or cohort studies analysed as case-control) on the relation between VDT exposure during pregnancy and the outcome were sought by reviewing reference lists in relevant reports and by conducting manual and computer searches of the reports published in English. MEASUREMENTS AND MAIN RESULTS: The nine reviewed reports included information on about 9000 cases of spontaneous miscarriages, 1500 of low birth weight, 2000 of congenital malformation, and 50,000 controls. The results of these studies on each outcome of pregnancy examined were reduced to a single 2 x 2 table (cases/controls--exposed/unexposed). Pooled odds ratio (OR) estimates were computed separately for miscarriage, low birth weight, and congenital malformation. Seven studies analysed the relation between VDT exposure in pregnancy and the risk of miscarriage: the estimates' crude OR of spontaneous abortion ranged from 0.9-1.2 and the pooled OR was 1.0 (95% confidence interval (CI) 0.9, 1.0). No consistent evidence of increasing risk with duration of exposure to VDT was found. Two studies analysed the relation between VDT use and risk of having a low birth weight infant: the OR estimates in the individual studies were 1.0 and 1.1. Likewise, no relation emerged from the five studies providing information on congenital malformations and VDT use: the pooled OR was 1.0 (95% CI 0.9, 1.2). No specific malformation pattern emerged. CONCLUSIONS: This meta-analysis provides reassuring evidence on the absence of any major risk of adverse pregnancy outcome as a result of exposure to a VDT. With the number of cases reviewed, it was possible to exclude excess risk of 20% for spontaneous abortion, low birth weight, and congenital malformations. PMID- 8228760 TI - Community wide outbreak of hepatitis A linked to children in day care centres and with increased transmission in young adult men in Florida 1988-9. AB - STUDY OBJECTIVE: To investigate a community wide outbreak of hepatitis A (HA). DESIGN: Description of the outbreak, with a case-control study to assess transmission. SETTING: A Florida county, USA, 1988-9. SUBJECTS: A total of 311 cases of HA. MEASUREMENTS AND MAIN RESULTS: A 13 month outbreak of HA is described. Most of the 311 cases (95%) were residents of a large metropolitan area (attack rate per 10,000 population (AR) = 3.7) and two smaller cities (AR = 61.5 and AR = 6.4). The ARs were greater for males than females and for residents aged 25-34 years (9.7) and < 5 years (8.3). Altogether 37% of cases were linked to day care centres, independent of the city of residence. A household case control study showed an increased risk of HA in households in which a child attended a day care centre (p = 0.02), and centres that could take more than 50 children had an increased risk of HA introduction than smaller ones (p = 0.05). CONCLUSIONS: Day care centres were an important source of HA in the community, and the need for timely surveillance and immunoglobin prophylaxis is emphasised. Homosexual transmission may have played an important role in this outbreak. PMID- 8228761 TI - Social epidemiology of chickenpox in two British national cohorts. AB - STUDY OBJECTIVE: To provide a quantitative description of factors independently predictive of reported chickenpox infections in two national cohorts of British children. DESIGN: Longitudinal cohort study design employing logistic regression analysis of data obtained in the 1970 British Births Survey (later to become the Child Health and Education Study, CHES), and the 1958 British Perinatal Mortality Survey (later to become the National Child Development Survey, NCDS). SETTINGS: One-week birth cohorts covering the whole of the United Kingdom. PARTICIPANTS: Data were obtained from questionnaires administered to the carers of 10,196 children born in the UK between 5 and 11 April 1970 (CHES) and 10,927 children born in the UK between 3 and 9 March 1958 (NCDS). These numbers consist of the whole of the surviving cohorts excluding those for whom data were incomplete. MEASUREMENTS: Biological, social, and medical factors in the parents and children, as recorded by the child's principle carer or from clinical notes. MAIN RESULTS: Chickenpox by the age of 10 years was reported to be more common in the children of advantaged families (higher social class, higher parental education levels), with a higher prevalence in those parts of the United Kingdom normally associated with affluence, such as the South East and South West of England, and lower rates in Wales and Scotland. Chickenpox by 10 years was also associated with more crowding in the home. A similar but less marked pattern occurred for chickenpox by the age of 11 years in the 1958 NCDS cohort. This social distribution apparently reflected overall rather than age-specific susceptibility. CONCLUSIONS: The national and international pattern of chickenpox epidemiology indicate that both social and climatological factors may be important in defining groups at risk. Further research is indicated if a vaccination service is to be implemented in this country. PMID- 8228762 TI - Increased wheeze but not bronchial hyperreactivity near power stations. AB - STUDY OBJECTIVE: In a previous study a higher than expected prevalence of asthma was found in Lake Munmorah, a coastal town near two power stations, compared with another coastal control town. This study aimed to compare atopy, bronchial hyperreactivity, and reported symptoms of asthma in the power station town and a second control area with greater socioeconomic similarity. DESIGN: A cross sectional survey was undertaken. SETTINGS: Lake Munmorah, a coastal town near two power stations, and Dungog, a country town in the Hunter Valley, NSW, Australia. PARTICIPANTS: All children attending kindergarten to year 6 at all schools in the two towns were invited to participate in 1990. The response rates for the questionnaire for reported symptoms and associated demographic data were 92% in Lake Munmorah and 93% in Dungog, with 84% and 90% of children respectively being measured for lung function, atopy, and bronchial reactivity. There were 419 boys and 432 girls aged 5 to 12 years. MEASUREMENTS AND MAIN RESULTS: Main outcome measures were current wheeze and bronchial hyper-reactivity, defined as a fall in forced expiratory volume in 1 second (FEV1) or peak expiratory flow (PEF) of 20% or more. Current wheeze was reported in 24.8% of the Lake Munmorah children compared with 14.6% of the Dungog children. Bronchial hyper-reactivity was similar for both groups--25.2% in Lake Munmorah and 22.3% in Dungog. The mean baseline FEV1 was lower in Lake Munmorah than in Dungog (p < 0.001). Dungog children were more likely to have positive skin test to house dust mite (Dungog 27.0%, Lake Munmorah 20.2%, p = 0.028) but there were no other differences in skin test atopy in the two towns. After adjusting for age, gender, any smoker in the house, and positive dust mite skin test, the odds of current wheeze in Lake Munmorah compared with Dungog was 2.16 (95% confidence interval 1.45, 3.15). CONCLUSIONS: Baseline lung function was lower and reported symptoms of asthma were higher in the power station town, but bronchial hyper-reactivity and skin test defined atopy were similar in the two communities. These results are consistent with the previous study and confirm the increased presence of reported symptomatic illness in the town near power stations. PMID- 8228763 TI - Ovarian cancer and ABO blood groups. AB - OBJECTIVE: To determine whether the distribution of ABO blood groups in women with ovarian cancer differs from that in the general population in a large, defined English region. DESIGN: Analysis of record abstracts of hospital care held in the Oxford record linkage study supplemented with data from the Oxford cancer registry. SETTING: Oxford Regional Health Authority area. SUBJECTS: A total of 1261 women who had ovarian cancer between 1968 and 1986 with ABO blood groups recorded on the Oxford Record Linkage Study and cross checked against the cancer registry comprised the study group. MEASUREMENTS AND MAIN RESULTS: The relative incidence of A:O and B:O blood groups in women with ovarian cancer were compared with the general population in the same region. Ovarian cancer was more common in women of blood group A than in others, with a relative incidence of 1.17. In particular, adenocarcinomas were the most common type of tumour and were associated with blood group A. The association was more striking in married women than in single women probably reflecting differences associated with parity. CONCLUSION: The association between ABO blood groups and ovarian cancer found in this English population is similar in size to that reported from several other populations. Childbearing is known to reduce the risk of ovarian cancer and our findings suggest that the blood group association may be most apparent in married, parous (that is, relatively low risk) women. PMID- 8228764 TI - Epidemiological trend in the distribution of cancer in Kashmir Valley. AB - STUDY OBJECTIVE: To study the distribution of cancers, with particular emphasis on oesophageal cancer. DESIGN: Patient records for 1986-88 in the Department of Oncology, SK Institute of Medical Sciences, were searched to identify cases and types of cancer. SETTING: The Valley of Kashmir, India. SUBJECTS: Patients with cancer resident in the valley. MAIN RESULTS: The seven most common types of cancer in men were those of the oesophagus, lungs, stomach, skin, upper respiratory tract, and urinary tract. In women the most common types of cancer were those affecting the oesophagus, breast, cervix, stomach, skin, colon/rectum, and lungs. Cancer of the oesophagus was the most frequent type in both sexes, accounting for 42.9% of all types of cancer in the valley. This distribution of cancer types is strikingly different from that in the rest of India where oropharyngeal cancer is the most common form. Again, contrary to the trend in India as a whole, cervical cancer is not a leading type of cancer in the valley and is less frequent even than cancer of the breast in the women. CONCLUSIONS: The preponderance of oesophageal cancer was attributable to the local practice of drinking boiling hot salt tea. Universal male circumcision in the majority community in the valley was considered to be partly responsible for low cervical cancer frequency. PMID- 8228765 TI - Death certification after a diagnosis of presenile dementia. AB - STUDY OBJECTIVES: To assess the value of death certification for the epidemiological study of dementia, the frequency with which the condition was recorded on death certificates of patients diagnosed with some form of dementia before the age of 65 years was studied. A further objective was to identify variables associated with failure to record dementia on the certificate. DESIGN: A cohort of patients with presenile dementia, differentiated by a clinical algorithm applied to hospital case records, was traced through the National Health Service Central Registry and details of certified causes of death were obtained. SETTING: The Northern Regional Health Authority in England. SUBJECTS: Prevalent cases of presenile dementia resident in the northern health region during 1986 traced up to April 1992. MEASUREMENTS AND MAIN RESULTS: The underlying cause of death was recorded as dementia or as Alzheimer's disease in 53% of cases of clinically diagnosed presenile Alzheimer's disease, 33% of cases of presenile vascular dementia, and 10% of cases of presenile dementia secondary to another neurological condition. Dementia or Alzheimer's disease was recorded in any part of the certificate in 75% of cases of Alzheimer's disease, 52% of vascular dementia, 33% of other dementias, and in 65% of cases overall. Dementia or a cerebral condition of a kind that can result in dementia was recorded in 80% of all cases. Failure to mention dementia was related to the clinical type of dementia, shorter duration of illness, and earlier period of study. CONCLUSIONS: The underlying cause of death seriously understates the frequency of dementia, but when the recording of other brain disease is taken into account the presence of potentially dementing brain disease is recorded much more frequently. It is suggested that coding chronic conditions present at death, such as dementia, in addition to those causing or contributing to death would improve the value of death certificates for epidemiological purposes. PMID- 8228766 TI - Mortality of frail elderly people living at home in relation to housing conditions. AB - STUDY OBJECTIVE: The study was undertaken to identify any relationship between the mortality of frail elderly people living at home and their housing conditions. DESIGN AND SETTING: A cohort of frail elderly people living in Yao City, Osaka Prefecture, Japan was followed for six years. Data on demographic variables, activities of daily living, the disease that was the main cause of the disability, medical treatment, the feasibility of continuing family care, and some housing conditions were collected in December 1983 by interview during home visits. Deaths were checked for each year for five years. SUBJECTS: Altogether 423 people aged 65 years or more living at home in Yao City who were disabled and housebound for at least three months were studied. MEASUREMENTS AND MAIN RESULTS: Life table analysis and multivariate logistic regression analysis were done. The cumulative survival rates of old people with good housing conditions were higher than those with poor housing conditions in each group stratified by age, sex, or the activities of daily living level. The odds ratio for poor housing conditions compared with good housing conditions for five year mortality remained highly significant after adjustment for potential confounding factors including age, sex, and the activities of daily living level by logistic regression analysis. CONCLUSIONS: The mortality of frail elderly people living at home is affected by housing conditions. Programmes to improve housing conditions of the disabled are important for home care of the elderly. PMID- 8228767 TI - Coronary heart disease risk factors in sub-Saharan Africa: studies in Tanzanian adolescents. AB - STUDY OBJECTIVE: To assess the level of cardiovascular risk factors in young people in sub-Saharan Africa living in rural and urban settings. DESIGN: Cross sectional survey of the population aged 15 to 19 years. SETTING: Eight rural Tanzanian villages in three regions, and two districts in Dar es Salaam. PARTICIPANTS: 664 males and 803 females in rural villages and 85 males and 121 females in the city. Response rates for total population were 74% to 94% in the rural areas and 60% in the city. MEASUREMENTS AND RESULTS: Measurements included blood pressure, body mass index, serum lipids, and blood glucose concentrations (fasting and two hours after 75 g glucose). Blood pressure was slightly but significantly higher in young women than in young men (115/67 mmHg versus 113/65 mmHg) and increased significantly with age. Only 0.4% subjects had blood pressure greater than 140 and/or 90 mmHg. There were no urban-rural differences. Body mass index was higher in females (mean (SD) 20.3 (2.8) kg/m2) than males (18.5 (2.1)). Overweight was found in only 0.6% at age 15 years but 5.4% at age 19 years. Serum cholesterol concentrations were low at 3.5 mmol/l in males and 3.7 mmol/l in females. Only 7% had values above 5.2 mmol/l. The highest concentrations were found in the city and in Kilimanjaro, the most prosperous rural region. Serum triglycerides were 1.0 (0.5) mmol/l in males and 1.1 (0.5) mmol/l in females, and were highest in the city dwellers. Diabetes was rare (0.28% males, 0.12% females) but impaired glucose tolerance was present in 4.7% and 4.1% respectively. Drinking alcohol was equally prevalent in males and females, reaching 30% at age 19 years. Only 0.4% of females smoked compared with 7.3% of males. Smoking was commoner in rural areas that in the city. CONCLUSIONS: Several risk factors for cardiovascular disease were found in Tanzanian adolescents, but levels were much lower than in studies reported from developed nations. The challenge is to maintain these low levels as the population becomes more urbanised and more affluent. PMID- 8228768 TI - Alcohol consumption and the risk of acute myocardial infarction in women. AB - STUDY OBJECTIVE: To investigate the relationship between alcohol consumption and the risk of acute myocardial infarction in women. DESIGN: This was a hospital based, case-control study carried out between 1983 and 1990. Main outcome measures were average daily number of drinks of various alcoholic beverages consumed and corresponding multivariate relative risk estimates and 95% confidence intervals (CI). SETTING: A network including major teaching and general hospitals in northern Italy. SUBJECTS: Cases were 298 women with acute myocardial infarction but no history of ischaemic heart disease and controls 685 women admitted to hospital for acute conditions, unrelated to alcohol consumption or to known or suspected risk factors for ischaemic heart disease. MEASUREMENTS AND MAIN RESULTS: Compared with non-drinkers, the estimated relative risks (RR) were 0.7 (95% CI 0.5, 1.0) for one drink or less per day, 0.8 (95% CI 0.6, 1.2) for more than one to two drinks per day, 1.4 (95% CI 0.8, 2.3) for more than two to three, and 2.6 (95% CI 1.5, 4.6) for more than three drinks per day. These estimates were consistent across strata of selected covariates, including age, education, and smoking. Allowance for major identified risk factors for myocardial infarction did not materially modify the risk estimate for light drinkers (RR 0.7, 95% CI 0.5, 1.1), but reduced the RR in heavy drinkers to 1.8 (95% CI 0.9, 3.5). CONCLUSIONS: This study indicates that women who do not drink alcohol have a risk of myocardial infarction that is higher than that of light drinkers, although the protection of light drinking was not significant. Among drinkers, however, there was a significant direct trend in risk with dose. The raised risks in heavy drinkers may reflect a real association or result from other unfavourable characteristics or habits associated with high alcohol consumption. PMID- 8228769 TI - Differences in priorities assigned to patients by triage nurses and by consultant physicians in accident and emergency departments. AB - STUDY OBJECTIVES: To investigate whether the greater urgency assigned to accident and emergency patients by triage nurses than by accident and emergency doctors was uniform across all patient groups. DESIGN: Patients attending an accident and emergency department between 8.00 am and 9.00 pm over a six week period were assessed prospectively for degree of urgency by triage nurses, and retrospectively for urgency by one of two consultant accident and emergency doctors. Patients were grouped according to their clinical mode of presentation. SETTING: An accident and emergency department of a district general hospital in the Midlands, UK, in 1990. PATIENTS: 1213 patients who presented over six weeks. MEASUREMENTS AND MAIN RESULTS: As might be expected, patients' conditions were assessed as being more urgent prospectively than retrospectively. This finding, however, was not uniform across all patient groups. Nurses' assessments of urgency tended to favour children and patients who presented with eye complaints and gave less priority to medical cases, particularly those with cardiorespiratory symptoms. CONCLUSIONS: These findings have implications for all those involved in the organisation of triage systems and in the training of nurses in accident and emergency departments. It is essential that judgements on how urgently patients need to be seen are made in a completely objective manner. PMID- 8228770 TI - Computerised linking of medical records: methodological guidelines. AB - OBJECTIVES: To report on the development of computer assisted methods for linking medical records and record abstracts. DESIGN: The methods include file blocking, to put records in an order which makes searching efficient; matching, which is the process of comparing records to determine whether they do or do not relate to the same person; linkage, which is the process of assembling correctly matched records into a time sequenced composite record for the individual; and validation checks and corrections, in which any inconsistencies between different records for the same person are identified and corrected. SETTING: The dataset comprising the Oxford record linkage study which includes hospital inpatient records and vital records. RESULTS AND CONCLUSIONS: Probability matching, using an array of identifiers, achieves much higher levels of correct matching than is generally achievable by exact character by character comparisons. The increasing use of information technology to store data about health and health care means that there is increasing scope to link records for research and for patient care. Sophisticated methods to achieve this on a large scale are now available. PMID- 8228771 TI - Case-control studies in cancer patients as a surveillance system of occupational exposure in the European Community. European Community Working Party. AB - STUDY OBJECTIVE: The main aim was to detect known relationships between lung and blood cancers and various occupational exposures (using job titles as proxies) using a case-control design. The suitability of this system for routine surveillance could then be assessed. DESIGN: A case-control study was carried out in 1989. SETTING: Hospitals in eight European Community countries. SUBJECTS: Men aged 25 to 75 years with incident and prevalent cancer of the lung (190 cases), haematopoietic system (210 cases), or gastrointestinal tract (245 controls) were studied. MEASUREMENTS AND MAIN RESULTS: The crude estimate of the overall odds ratio exposure (OR) for relevant occupational exposure of lung cancer relative to gastrointestinal cancer was 1.20 (95% confidence interval (CI) 0.82, 1.77). In a logistic regression analysis adjusting for country, age at diagnosis, smoking, and alcohol consumption, the overall OR was not greatly changed. A significant interaction of occupational exposure and age at diagnosis showed that lung cancer patients diagnosed at a younger age had a higher OR than patients diagnosed at an older age. Thus, the overall, insignificant result may have been due to a low reliability of occupational history in older age or to a selective mechanism related to age. The overall OR for occupational exposure of cancer of the blood relative to gastrointestinal cancer was 0.88 (95% CI 0.60, 1.31). The logistic regression analysis did not alter these results. CONCLUSION: A surveillance based on a case-control design using job titles would not be sensitive enough to detect possible occupational risks. PMID- 8228772 TI - Variability of self reported measures of alcohol consumption: implications for the association between drinking in pregnancy and birth weight. AB - STUDY OBJECTIVE: To describe the intra-subject variability of self reported maternal alcohol consumption using different ways of collecting information and to analyse the implications of this variability for research into the effect of low to moderate maternal alcohol consumption on birth weight. DESIGN: This was a longitudinal study. Self reported maternal alcohol consumption before, during, and after pregnancy was assessed on four occasions over two years. The data were collected by two self administered questionnaires and during two personal interviews (one by phone and another face to face). SETTINGS: The Obstetrics Department, Odense University Hospital, Odense, Funen, Denmark. PARTICIPANTS: A total of 2880 pregnant women were recruited consecutively from the hospital catchment area. Altogether 328 pregnant women and their babies were selected. All women who reported an average alcohol consumption of five drinks or more per week were recruited to the study (164 women) and a 1:1 control group was selected from the remaining women based upon two matching criteria: expected date of delivery and the women's year of birth. Some 279 women (85%) completed the study. MEASUREMENTS AND MAIN RESULTS: Self reported alcohol consumption (number of drinks per week) and birth weight (g) were the main outcomes. Women's self reported alcohol consumption varied over time and according to the data collection method. When different methods of data collection were used to assess alcohol intake in similar periods of time, significant differences in reporting were found despite the relatively high correlations between the measurements. Although a consistent reduction in birth weight with increasing consumption of alcohol was found, there were differences in the shape and strength of this association when comparing the six available alcohol measurements. CONCLUSIONS: The type of questions used, the way the data were collected, the period of time referred to, and the time the questions were asked, should be taken into consideration when describing the drinking pattern of pregnant women. Furthermore, birth weight results from studies that have used different alcohol measures should be interpreted or compared with caution because of possible large differences resulting from the differing methods of assessing fetal exposure to alcohol. PMID- 8228773 TI - Intestinal transit time in the population calculated from self made observations of defecation. AB - STUDY OBJECTIVES: To assess the feasibility of estimating intestinal transit time in the general population using self recorded data on stool form, frequency of defecation, and the interdefecatory time interval. DESIGN: Prospective measurement of bowel function. SETTING: Bristol, Avon, UK between 1987 and 1989. SUBJECTS: Subjects were drawn from 1897 people who comprised 72.2% of a stratified random sample of all men aged 40-69 years and women aged 25-69 years on the lists of 19 general medical practitioners. Altogether 1561 subjects (59.4%) recorded bowel function and a subsample of 98 (50 women and 48 men) had intestinal transit time measured. MEASUREMENTS AND MAIN RESULTS: The interdefecatory time interval and stool form (on a validated 1-6 scale sensitive to transit time) were recorded prospectively from three consecutive defecations. In the subsample the mean intestinal transit time was measured simultaneously using a four marker, two stool x ray technique. Multiple regression analysis was used to assess the extent to which intestinal transit time could be predicted from the defecatory data. The formulas obtained were then applied to the whole study population. In women, intestinal transit time was best predicted by the formula 103-1.23 (DF)--4.69 (SFS)+0.638 (IDTI), where DF is the stated defecation frequency per week, IDTI is the interdefecatory time interval, and SFS is the sum of the three stool form scores, for which the correlation coefficient r = 0.736. For men the intestinal transit time = 79-1.33 (DF)--1.88 (SFS)+0.329 (IDTI), for which the correlation coefficient r = 0.541. The predicted intestinal transit time was longer in women than men at equivalent ages. Women of childbearing age had longer transit times than older women. CONCLUSIONS: Observations made by untrained subjects can be used to estimate intestinal transit time in epidemiological studies. A gender related difference in transit time exists. PMID- 8228774 TI - Impact of a postcard versus a questionnaire as a first reminder in a postal lifestyle survey. AB - STUDY OBJECTIVE: The study aimed to consider the impact of two different types of reminder on response rates and costs in a postal survey. DESIGN: The study was a cross sectional survey. A self-completion lifestyle questionnaire was used. Those who did not respond after the initial mailing were randomly allocated to receive either a postcard or questionnaire as a first reminder. All outstanding non responders received a questionnaire as a second reminder. SUBJECTS: A representative sample of 698 adults aged 16-70 was used, drawn from a family health services authority register. MAIN RESULTS: Postcard reminders were as effective as questionnaire reminders in increasing response whether one or two reminders are sent. The costs per response were calculated. Two questionnaires as reminders were found to be 1.7 times more expensive than a postcard plus questionnaire. Including the initial mailing, the cost per response using all questionnaires was 1.3 times the cost when a postcard was used for the first reminder. CONCLUSIONS: To increase the response to a postal survey effectively and economically, two reminders should be sent--first a postcard and then a questionnaire. PMID- 8228775 TI - Mortality among drug injectors and notified addicts. PMID- 8228776 TI - Selective migration by birthweight. PMID- 8228777 TI - Ion channel activity in lobster skeletal muscle membrane. AB - Ion channel activity in the sarcolemmal membrane of muscle fibers is critical for regulating the excitability, and therefore the contractility, of muscle. To begin the characterization of the biophysical properties of the sarcolemmal membrane of lobster exoskeletal muscle fibers, recordings were made from excised patches of membrane from enzymatically induced muscle fiber blebs. Blebs formed as evaginations of the muscle sarcolemmal membrane and were sufficiently free of extracellular debris to allow the formation of gigaohm seals. Under simple experimental conditions using bi-ionic symmetrical recording solutions and maintained holding potentials, a variety of single channel types with conductances in the range 32-380 pS were detected. Two of these ion channel species are described in detail, both are cation channels selective for potassium. They can be distinguished from each other on the basis of their single channel conductance and gating properties. The results suggest that current flows through a large number of ion channels that open spontaneously in bleb membranes in the absence of exogenous metabolites or hormones. PMID- 8228778 TI - Kinematics of birdsong: functional correlation of cranial movements and acoustic features in sparrows. AB - The movements of the head and beak of songbirds may play a functional role in vocal production by influencing the acoustic properties of songs. We investigated this possibility by synchronously measuring the acoustic frequency and amplitude and the kinematics (beak gape and head angle) of singing behavior in the white throated sparrow (Zonotrichia albicollis) and the swamp sparrow (Melospiza georgiana). These birds are closely related emberizine sparrows, but their songs differ radically in frequency and amplitude structure. We found that the acoustic frequencies of notes in a song have a consistent, positive correlation with beak gape in both species. Beak gape increased significantly with increasing frequency during the first two notes in Z. albicollis song, with a mean frequency for note 1 of 3 kHz corresponding to a gape of 0.4 cm (a 15 degrees gape angle) and a mean frequency for note 2 of 4 kHz corresponding to a gape of 0.7 cm (a 30 degrees gape angle). The relationship between gape and frequency for the upswept third note in Z. albicollis also was significant. In M. georgiana, low frequencies of 3 kHz corresponding to beak gapes of 0.2-0.3 cm (a 10-15 degrees break angle), whereas frequencies of 7-8 kHz were associated with flaring of the beak to over 1 cm (a beak angle greater than 50 degrees). Beak gape and song amplitude are poorly correlated in both species. We conclude that cranial kinematics, particularly beak movements, influence the resonance properties of the vocal tract by varying its physical dimensions and thus play an active role in the production of birdsong. PMID- 8228779 TI - The active control of wing rotation by Drosophila. AB - This paper investigates the temporal control of a fast wing rotation in flies, the ventral flip, which occurs during the transition from downstroke to upstroke. Tethered flying Drosophila actively modulate the timing of these rapid supinations during yaw responses evoked by an oscillating visual stimulus. The time difference between the two wings is controlled such that the wing on the outside of a fictive turn rotates in advance of its contralateral partner. This modulation of ventral-flip timing between the two wings is strongly coupled with changes in wing-stroke amplitude. Typically, an increase in the stroke amplitude of one wing is correlated with an advance in the timing of the ventral flip of the same wing. However, flies do display a limited ability to control these two behaviors independently, as shown by flight records in which the correlation between ventral-flip timing and stroke amplitude transiently reverses. The control of ventral-flip timing may be part of an unsteady aerodynamic mechanism that enables the fly to alter the magnitude and direction of flight forces during turning maneuvers. PMID- 8228780 TI - The cost of hovering and forward flight in a nectar-feeding bat, Glossophaga soricina, estimated from aerodynamic theory. AB - Energy expenditure during flight in animals can best be understood and quantified when both theoretical and empirical approaches are used concurrently. This paper examines one of four methods that we have used to estimate the cost of flight in a neotropical nectar-feeding bat Glossophaga soricina (Phyllostomidae), namely the use of kinematic and morphological data and aerodynamic theory to estimate the mechanical power requirements (power output) for hovering and horizontal forward flight. A hot-wire anemometer was used to measure induced velocity (the velocity of air accelerated by the wings) during hovering in order to estimate induced power. Our estimate of aerodynamic power (the sum of induced, profile and parasite powers) required for a 0.0105 kg G. soricina to hover is 0.15 W and our estimate of the inertial power (the power required to oscillate the wings) is 0.19 W. Thus, the total mechanical power for hovering is 0.34 W or 32.4 W kg-1. The mechanical power required for horizontal forward flight, near the minimum power flight speed (4.2 ms-1) for a 0.0117 kg bat is 0.14 W (12.3 W kg-1), of which 0.10 W is aerodynamic power and 0.042 W is inertial power. Comparison with our results on metabolic power requirements estimated from nectar intake gives a mechanical efficiency of 0.15 for hovering flight and 0.11 for forward flight near the minimum power speed. PMID- 8228781 TI - Actions of vesamicol on an alpha-bungarotoxin-sensitive neuronal nicotinic acetylcholine receptor. AB - Electrophysiology and binding studies were used to determine the actions of vesamicol [2-(4-phenylpiperidino)cyclohexanol (AH5183)] on an alpha-bungarotoxin sensitive, neuronal nicotinic acetylcholine receptor in the nervous system of the cockroach, Periplaneta americana. Electrophysiological studies on an identified motor neurone revealed a reversible blocking action of (+/-)-vesamicol on the response to ionophoretically applied acetylcholine with an IC50 value of 8.0 x 10(-6) mol1(-1). The block was weakly voltage-dependent over the membrane potential range of -50 mV to -90 mV, and appeared to be non-competitive. No difference in potency was observed between the resolved stereoisomers. (+/-) Vesamicol was found to suppress specific binding of 125I-labelled alpha bungarotoxin to cockroach nervous tissue with an IC50 value of 5.1 x 10(-3) mol1( 1) and an estimated Hill coefficient of 0.73. Differences in the Hill coefficients were found when the resolved stereoisomers were tested separately. These data provide the first demonstration of a blocking action by vesamicol of a neuronal nicotinic acetylcholine receptor. PMID- 8228782 TI - The behavioural state of satiation in the leech is regulated by body distension and mimicked by serotonin depletion. PMID- 8228783 TI - Glibenclamide binding in vertebrate cardiac membranes. PMID- 8228784 TI - Maximum metabolism and the aerobic factorial scope of endotherms. AB - Minimum and maximum metabolism in response to cold were measured in 30 species of Australian monotremes, marsupials, eutherians and birds. In marsupials and the echidna, maximum metabolism was also determined during treadmill locomotion. These data were used to determine, for the first time, the relationships between maximum metabolism and body mass in the four endothermic groups and to compare aerobic factorial scopes (the ratio of maximum to minimum metabolism) elicited by cold and locomotion. The effect of body mass on maximum metabolism is the same in marsupials and eutherians (the therians) but is significantly less in birds. At the same body mass, there is no difference between the two therian groups for either minimum or maximum metabolism induced by either cold or locomotion. Aerobic scope during cold is significantly higher in marsupials (8.3) than in eutherians (5.1), birds (5.4) and monotremes (5.4). Aerobic scope during locomotion in all groups is almost twice that observed in cold conditions. PMID- 8228785 TI - Sensory and neurosecretory innervation of leech nephridia is accomplished by a single neurone containing FMRFamide. AB - The neural control of the excretory system of the medicinal leech Hirudo medicinalis has been characterized morphologically and chemically using light and electron microscopy, immunocytochemistry and biochemistry. Immunoreactivity against RFamide-like peptides revealed elaborate neuronal aborizations of a neurone in the nephridium, around the urinary bladder sphincter and in the central nervous system. The processes arose from the nephridial nerve cell (NNC), a previously identified receptor neurone. Using a combination of reverse-phase high pressure liquid chromatography, radioimmunoassay and subsequent Edman degradation and mass spectrometry, authentic FMRFamide has been identified as the major peptide of the NNC. Sensory and neurosecretory innervation of the nephridia is thus accomplished by a single neurone, which is thought to modulate nephridial performance. PMID- 8228786 TI - Genetics of non-insulin-dependent diabetes mellitus (NIDDM) PMID- 8228787 TI - Hepatitis B virus precore and core gene mutations in chronic hepatitis B and hepatocellular carcinoma patients. PMID- 8228788 TI - Lipoprotein(a) and treatment of chronic renal disease. AB - OBJECTIVES: To compare lipoprotein(a) [Lp(a)] and albumin concentrations in patients with chronic renal disease receiving different forms of treatment and to determine, if any, the relationship between these variables. DESIGN: A prospective cross-sectional, case-controlled study. SETTING: A tertiary referral nephrology and dialysis unit. SUBJECTS: Forty-four consecutive non-diabetic patients with chronic renal failure treated by renal transplantation (n = 18), haemodialysis (n = 18), continuous ambulatory peritoneal dialysis (CAPD; n = 8), and 30 healthy controls from subjects drawn from University personnel were studied. INTERVENTIONS: Fasting morning venous blood was analysed for Lp(a), albumin, total cholesterol and glucose concentrations. MAIN OUTCOME MEASURES: Comparison of plasma levels of these variables between the sub-groups. RESULTS: Concentrations (median; 95% CI) of Lp(a) were significantly (P < 0.05) higher (38.4 mg dl-1; range 15.4-72.0) and of albumin lower (31.6 g l-1; range 28-35.2) in the CAPD group compared with both control subjects and other groups of chronic renal disease patients. CONCLUSIONS: The elevated Lp(a) concentrations seen only in association with reduced albumin concentrations in CAPD patients suggest a regulatory role for albumin with albumin losses stimulating production of Lp(a). PMID- 8228789 TI - Follow-up study of patients with clinically suspected deep venous thrombosis and a normal venogram. AB - OBJECTIVES: To evaluate the clinical course in patients with clinically suspected deep venous thrombosis (DVT) of the leg and a normal venogram. DESIGN: Prospective study over 15 months with a follow-up of 4-12 (median 8.6) months after a normal venogram. A questionnaire survey was performed at follow-up. Information from general practitioners and medical records was reviewed. An alternative diagnosis was established at presentation and at the time of follow up. SETTING: The Department of Internal Medicine in a Danish university hospital. SUBJECTS: A total of 133 consecutive out-patients referred with clinical suspicion of DVT and a normal venogram. MAIN OUTCOME MEASURES: The state of symptoms at follow-up. The frequency of referrals to hospitals and contacts with general practitioners or medical specialists in the follow-up period. Clinical diagnoses provided at presentation and at follow-up. RESULTS: The follow-up response rate was 78% (n = 104). The symptoms were still present at follow-up in 53 (51%) patients. More than half of the patients had been referred to medical facilities for the same disorder. Diagnoses could be established in 93 (70%) of the 133 patients at presentation and in 119 (89%) at follow-up. CONCLUSIONS: The majority of patients with clinical signs and symptoms of a DVT and a normal venogram may require a follow-up surveillance programme to ensure correct diagnosis and adequate treatment. Further studies are recommended to confirm our results and to assess the cost-effectiveness. PMID- 8228790 TI - Altered adrenocorticotropin and cortisol secretion in abdominal obesity: implications for the insulin resistance syndrome. AB - OBJECTIVES: To investigate the relationship between the pituitary-adrenocortical function, abdominal obesity, and insulin resistance syndrome. DESIGN: A prospective study. SETTING: Helsinki University Hospital, Finland. SUBJECTS: Sixty-six healthy males aged 30-55 years. MAIN OUTCOME MEASURES: Insulin, C peptide, cortisol and ACTH responses during the oral glucose tolerance test (OGTT), and the cortisol response to dexamethasone suppression and intravenous adrenocorticotrophic hormone (ACTH) stimulation. RESULTS: The subjects in the highest tertile of the waist-to-hip ratio (WHR) had lower high-density lipoprotein cholesterol (HDLC) (P < 0.05), but higher triglyceride (TG), insulin, and C-peptide levels, ACTH response to glucose at 2 h, and cortisol response to ACTH (P < 0.01) than those in the lowest tertile. The cortisol response to ACTH correlated positively, but cortisol levels during the OGTT correlated negatively with WHR. The ratio of these cortisol determinations correlated positively with the body-mass index (BMI) (r = 0.554; P < 0.001), WHR (r = 0.536; P < 0.001), TG (r = 0.397; P = 0.001), fasting insulin (r = 0.534; P < 0.001) and C-peptide (r = 0.458; P < 0.001), and negatively with HDLC (r = 0.353; P = 0.004). In multiple regression analyses, BMI and the 2-h ACTH response to glucose were significant predictors of WHR and, in addition, the cortisol ratio, WHR, and BMI of insulin. CONCLUSIONS: Abdominal obesity may be associated with subtle central adrenal insufficiency, which might also affect insulin and lipoprotein metabolism. PMID- 8228791 TI - Lipids and lipoproteins as risk factors for coronary heart disease in men with abnormal glucose tolerance: the Honolulu Heart Program. AB - OBJECTIVES: To evaluate lipids and lipoproteins as risk factors for coronary heart disease (CHD) in older men with non-insulin-dependent diabetes (NIDDM) or abnormal glucose tolerance compared with normoglycaemic men. DESIGN: A prospective, population-based cohort study based on the lipoprotein examination (1970-72) of the Honolulu Heart Program. Follow-up was through to December 1988. SETTING: Honolulu, Hawaii. SUBJECTS: Japanese-American men, ages 51-72 at baseline: 2042 with 1 h glucose < 12.5 mmol l-1 (normal group); 376 on oral hypoglycaemic agents or with 1 h glucose > or = 12.5 mmol l-1 after 50 g oral glucose challenge (abnormal glucose tolerance group). None had prevalent coronary heart disease (CHD) or stroke at baseline. MAIN OUTCOME MEASURES: Incident CHD: definite non-fatal myocardial infarction (MI) or fatal CHD. RESULTS: There were 221 incident cases in the normal group, and 65 in the abnormal glucose tolerance group. Total and high-density lipoprotein (HDL) cholesterol were significant predictors of incident CHD in men with NIDDM or abnormal glucose tolerance after controlling for age, body-mass index, systolic blood pressure, pack-years of cigarettes and alcohol consumption (P < 0.05). Total, low-density lipoprotein (LDL) and very-low-density lipoprotein (VLDL) cholesterol were significant predictors in normal men, and HDL cholesterol was of borderline significance. CONCLUSIONS: Abnormal lipids and lipoproteins are significant, independent predictors of CHD in subjects with NIDDM or abnormal glucose tolerance. Attention to lipid and lipoproteins as CHD risk factors should be part of clinical management of these patients. PMID- 8228792 TI - Different effects of two methods of low-density lipoprotein apheresis on the coagulation and fibrinolytic systems. AB - OBJECTIVE: Immunoadsorption (IMA) and dextran sulfate adsorption (DSA) are two methods for selective extracorporeal elimination of low-density lipoproteins which are known as LDL apheresis. Their influence on haemostasis until now is widely unknown. DESIGN: The effects of both LDL apheresis procedures on the coagulation and fibrinolytic systems were compared amongst five patients treated with IMA and four patients who received a DSA therapy. SUBJECTS: All patients with severe heterozygous familial hypercholesterolaemia were participants in a long-term LDL apheresis programme with treatments every 1-2 weeks. INTERVENTION: Combined anticoagulation with heparin and citrate in IMA, and also heparin exclusively in DSA were used for the extracorporeal circulation. MEASURES: Blood samples were taken immediately before and after a single LDL apheresis and five times during the weekly interval until the next therapy. RESULTS: DSA had a significantly greater effect on standard clotting tests than IMA at the end of plasma therapy despite identical dosages of heparin. DSA caused a considerable reduction of the coagulation factors V, VIII:C, vWF:Ag, XI, XII, and prekallikrein by 48-99% at the end of apheresis treatment whereas only factor VIII:C showed a marked decrease of 72% after IMA. All abnormalities of the global coagulation tests and of most clotting factors were restored 1 day after treatment in both procedures followed by a moderate rebound phenomenon of single coagulation factors during the next few days in IMA-treated patients. CONCLUSION: DSA exerts a more profound effect on the coagulation system than IMA by a substantial co-elimination of various clotting factors in addition to the desired removal of atherogenic lipoproteins. PMID- 8228793 TI - Diabetic lipohypertrophy treated with suction-assisted lipectomy. AB - OBJECTIVES: To investigate the effectiveness of liposuction as treatment for lipohypertrophy in insulin-treated diabetic patients. DESIGN: Open clinical study. SETTING: Norwegian National Hospital, Oslo. SUBJECTS: Five consecutive diabetic patients operated on for insulin-induced lipohypertrophy. INTERVENTIONS: Syringe-assisted liposuction under local anaesthesia. MAIN OUTCOME MEASURES: Change in contours assessed by pre- and postoperative photographs, and by the patients' and surgeon's evaluations according to a four-point graded scale. RESULTS: Good or excellent results were obtained in all patients. Small surface irregularities were seen in two patients who had large volumes of fat removed from the proximal anterior thighs. Apart from this no side-effects or complications occurred. CONCLUSIONS: Insulin-induced lipohypertrophy ('insulin tumours') can be treated successfully with liposuction. PMID- 8228794 TI - Haematocrit: a predictor of cardiovascular mortality? AB - OBJECTIVES: The main purpose of the study was to assess a possible association between haematocrit (Hct) and long-term cardiovascular disease (CVD) mortality. DESIGN: An extensive examination programme was carried out in 2014 men, defined as apparently healthy, during the period 1972 to 1975, including Hct measurements in a 25% random subsample. Sequential, cause-specific mortality was followed prospectively over a period of 16 years. SETTING: The survey was conducted at Medical Department B, Rikshospitalet, Oslo, Norway. SUBJECTS: The participants represented 86% of all eligible apparently healthy men working in five preselected companies in Oslo. INTERVENTIONS: No intervention was given by the study group during follow-up. MAIN OUTCOME MEASURES: Complete, cause-specific mortality figures after 8-16 years were obtained from the Norwegian Central Bureau of Statistics. RESULTS: Hct measures were obtained in 488 men (24.2%). Mean Hct was 47.2% (SD 2.9%). After correcting for differences in age, plasma cholesterol, systolic blood pressure, erythrocyte sedimentation rate and smoking habits (Cox proportional hazards model), an increase in Hct by 2 SDS was associated with an increase in CVD mortality by a factor ranging between 2.9 at 10, and 1.9 at 16 years (P < 0.05). A similar increased risk was observed earlier during follow-up but the number of deaths was too small for meaningful statistical analysis. No association was found between Hct and non-CVD mortality. CONCLUSIONS: Our data show that increased Hct is associated with an increased risk of dying from CVD--independent of conventional CVD risk factors. PMID- 8228795 TI - Complete membranous obstruction of the inferior vena cava: clinical characteristics of Chinese patients. AB - OBJECTIVES: To analyse the clinical presentations of membranous-type Budd-Chiari syndrome and the long-term outcome of balloon angioplasty. DESIGN: A case-control study. Patients who received abdominal sonography and were suspected of having hepatic venous outflow obstruction were advised to receive cardiac catheterization and angiography. SETTING: Patients were collected from Chung Hisao Hospital and Chang Gung Medical Centre. Angioplasty was done at Chang Gung Medical Centre. SUBJECTS: Seventeen patients who had the symptoms and signs of portal hypertension were suspected of having hepatic venous outflow obstruction after abdominal sonography and a radionuclide liver scan and eight of them were confirmed as having membranous-type obstruction. INTERVENTIONS: Simultaneous catheterization and contrast angiography was performed via the right brachial and femoral veins. Angioplasty was performed afterwards. MAIN OUTCOME MEASURES: The remission of clinical symptoms and signs and a decrease of the pressure gradient between the right atrium and inferior vena cava were used as indicators of successful intervention. RESULTS: All patients had remarkable remission of symptoms and signs. The average pressure gradient between right atrium and inferior vena cava dropped from 16.5 +/- 1.6 to 6.5 +/- 1 mmHg (mean +/- SD). CONCLUSION: The clinical presentations of membranous-type Budd-Chiari syndrome are somewhat different from hepatic vein occlusion. Balloon angioplasty may be a modality for curing this disease. A larger patient population is necessary to confirm this study. PMID- 8228796 TI - Distinguishing acyclovir neurotoxicity from encephalomyelitis. AB - OBJECTIVES: To define the clinical characteristics of acyclovir neurotoxicity and to outline how to distinguish it from viral encephalitis. DESIGN: Case series of acyclovir neurotoxicity. SETTING: All cases reported in Index Medicus or in bibliographic reviews of acyclovir neurotoxicity plus two representative studies of Varicella zoster and Herpes simplex virus encephalitis. SUBJECTS: Thirty-five patients who developed neuropsychiatric symptoms during acyclovir therapy. INTERVENTIONS: Analysis of the patients' demographics, risk factors, acyclovir dosages and duration, clinical and laboratory findings and outcome. MAIN OUTCOME MEASURES: All clinical and laboratory findings that were statistically significantly different from viral encephalitis. RESULTS: The median age was 53.3 years. The most common predisposing factors were the use of other potentially neurotoxic medications (17 cases) and acute or chronic renal failure (15 cases). Acyclovir levels were frequently found above the therapeutic range. The characteristic manifestations were confusion (15 cases), hallucination or delirium (9 cases), agitation (8 cases) and lethargy (10 cases). Few patients had associated tremors (11 cases). Fever, headache, seizures and focal neurologic findings were distinctly rare. Cerebrospinal fluid and computed tomography were normal except in patients with other central nervous system disorders. Symptoms appeared within 2 days of therapy in the majority and resolved completely within several days of discontinuing acyclovir. CONCLUSIONS: Acyclovir neurotoxicity is a self-limiting, dose-dependent phenomenon which is more common in the elderly, in patients with renal failure or in association with other neurotoxic medications. It is distinguished from viral encephalitis by its sudden onset, absence of fever or headache, lack of focal neurologic findings and normal cerebrospinal fluid. PMID- 8228797 TI - Streptokinase, but not tissue plasminogen activator, attenuates platelet aggregation in patients with acute myocardial infarction. AB - OBJECTIVES: To investigate if tissue plasminogen activator (tPA) and streptokinase given during acute myocardial infarction (AMI) have different effects on platelet aggregation which could contribute to the higher reocclusion rate observed after tPA. DESIGN: Open labelled on consecutive patients. SETTING: Coronary care unit. SUBJECTS: Twenty patients with chest pain and ST elevations on an electrocardiogram suggestive of AMI. INTERVENTIONS: Ten patients were treated with tPA (100 mg 3 h-1), 10 patients with streptokinase (1.5 x 10(6) IU 1 h-1). MAIN OUTCOME MEASURES: Before, immediately after and 24 h after fibrinolytic therapy, platelet aggregation was estimated with filtragometry and whole blood aggregometry. Fibrinogen, beta-thromboglobulin, elastase and the fibrinogen-derived peptide B beta 30-43 were also measured. RESULTS: The groups were comparable at baseline. Directly after treatment, streptokinase prolonged aggregation time in filtragometry with 112 +/- 140 s (P < 0.03) and reduced conductance in whole blood aggregometry by 6.2 +/- 6.1 omega (P < 0.03), both tests indicating inhibited platelet function. Fibrinogen decreased 2.5 +/- 1.0 g l-1 (P < 0.02). In the tPA-treated group corresponding changes were 68 +/- 225 s (NS) and 2.5 +/- 7 omega (NS) with no significant reduction in fibrinogen. After 24 h, at which time every patient was on acetylsalicylic acid, aggregation was inhibited in both groups as measured by aggregometry. Directly after fibrinolytic treatment, neutrophils were similarly activated in both groups with increments of elastase and B beta 30-43 by 26 +/- 46 micrograms l-1 (P < 0.03) and 280 +/- 381 pmol l-1 (P < 0.03) respectively (streptokinase) and by 12 +/- 6 micrograms l-1 (P < 0.02) and 919 +/- 856 pmol l-1 (P < 0.02) respectively (tPA). CONCLUSIONS: Despite similar degrees of platelet and leucocyte activation, streptokinase but not tPA treatment appears to inhibit platelet aggregation. One possible reason could be a streptokinase-induced pronounced decrease of fibrinogen and increase of fibrinogen split products. Therefore, further development of adjuvant antiplatelet therapy could be of clinical importance. PMID- 8228798 TI - Paraneoplastic polymyositis associated with a renal carcinoma. AB - A 56-year-old patient was admitted to hospital because of difficulties in swallowing and in opening his mouth. The serological transaminases were increased. Over the next 6 weeks a bioptically proven polymyositis (more proximally located) caused a respiratory insufficiency which had to be treated by artificial respiration. A tetraplegia connected with a large increase of creatine kinase was also seen. The general investigations were focused on malign processes with a particular search for bronchial carcinoma. At first a kidney cyst detected by ultrasound but was classified as harmless by several investigators. Finally, a computer tomogram of the abdomen demonstrated a cystic degenerated renal carcinoma, which had been cytologically tested also. The clinical course showed a typical aspect of a paraneoplastic syndrome: after extended nephrectomy a distinct regression of the musculary symptoms developed, but with the development of metastases the symptoms increased again. The interdependence of polymyositis and primary renal carcinoma would appear to be proven. PMID- 8228799 TI - Polyautoimmune syndrome in common variable immunodeficiency. AB - Common variable immunodeficiency (CVI) is a heterogenous disorder with hypogammaglobulinaemia and multiple bacterial infections primarily involving the sinopulmonary tract. CVI patients have been known to have an increased tendency to develop autoimmune manifestations. Commonly associated autoimmune diseases include immune thrombocytopenic purpura, autoimmune haemolytic anaemia, and rheumatoid arthritis. In this paper we report a case of CVI presenting with multiple unusual autoimmune diseases including parotitis, vitiligo, atrophic gastritis, pernicious anaemia, and primary biliary cirrhosis. To our knowledge, this is the first case of CVI with polyautoimmunity and antimitochondria antibody. Recognition of this association is important because early diagnosis and treatment can greatly influence the prognosis. PMID- 8228800 TI - Emergence of NK1.1+ cells as effectors of IFN-gamma dependent immunity to Toxoplasma gondii in MHC class I-deficient mice. AB - CD8+ T lymphocytes have been reported to play a major role in the protective immune response against acute infection with Toxoplasma gondii. In order to further assess the role of CD8+ cells in resistance against this protozoan we examined the ability of beta 2m-deficient mice, which fail to express MHC class I molecules and peripheral CD8+ lymphocytes, to survive tachyzoite challenge following vaccination with an attenuated parasite mutant. Surprisingly, vaccination of beta 2m-deficient mice induced strong resistance to lethal challenge, with > 50% surviving beyond 3 months. Vaccinated beta 2m-deficient mice, but not control heterozygotes, showed a five- to six-fold expansion in spleen cell number and approximately 40% of the splenocytes were found to express the NK markers NK1.1 and asialo GM1. Spleen cells from the vaccinated beta 2m deficient animals failed to kill either infected host cells or the NK target YAC 1. However, high levels of IFN-gamma were secreted when the cells were cultured in vitro with soluble T. gondii lysate, and this response was abolished by NK1.1+ but not CD4+ and CD8+ lymphocyte depletion, implicating the NK1.1+ population as the major source of IFN-gamma. More importantly, vaccine-induced immunity in beta 2m-deficient mice was completely abrogated by in vivo administration of antibody to NK1.1, asialo GM1, or IFN-gamma. Together, the data suggest that in class I deficient mice vaccinated against T. gondii, the absence of CD8+ effector cells is compensated for by the emergence of a population of NK1.1+ and asialo GM1+ cells which lack cytolytic activity, and that the protective action of these cells against the parasite is attributable to IFN-gamma production. The induction of this novel NK population may provide an approach for controlling opportunistic infections in immunocompromised hosts. PMID- 8228801 TI - Interleukin 10 (IL-10) upregulates functional high affinity IL-2 receptors on normal and leukemic B lymphocytes. AB - Interleukin 10 (IL-10) has recently been shown to induce normal human B lymphocytes to proliferate and differentiate into immunoglobulin (Ig)-secreting cells. Herein, we show that IL-10 also promotes DNA synthesis and IgM production by anti-CD40 activated B cell chronic lymphocytic leukemia (B-CLL). Most strikingly, IL-2 and IL-10 were found to synergize to induce the proliferation and differentiation of B-CLL cells. This synergy between IL-2 and IL-10 was also observed with normal B cells which proliferated strongly and secreted large amounts of IgM, IgG, and IgA. The observed synergy is likely to be due to the IL 10-induced increase of high affinity IL-2 receptors on both normal and leukemic B cells. This increase of high affinity receptor is associated to an increase of Tac/CD25 expression that can be detected by flow cytometric analysis. Taken together, these results indicate that IL-10 permits anti-CD40 activated B cells to respond to IL-2 through an induction of high affinity IL-2 receptors. This effect of IL-10 may partly explain how T cells, which activate B cells in a CD40 dependent fashion, induce B cell proliferation and differentiation mostly through IL-2. PMID- 8228802 TI - Two essential regulatory elements in the human interferon gamma promoter confer activation specific expression in T cells. AB - Like interleukin 2 (IL-2), interferon gamma (IFN-gamma) is an early response gene in T cells and both are prototypical T helper cell type 1 (Th-1) lymphokines. Yet IL-2 and IFN-gamma production are independently regulated, as demonstrated by their differential expression in certain T cell subsets, suggesting that the regulatory elements in these two genes must differ. To explore this possibility, the 5' flank of the human IFN-gamma gene was analyzed. Expression of IFN-gamma promoter-driven beta-galactosidase reporter constructs containing 538 bp of 5' flank was similar to that by constructs driven by the IL-2 promoter in activated Jurkat T cells; expression nearly as great was observed with the construct containing only 108 bp of IFN-gamma 5' flank. These IFN-gamma promoter constructs faithfully mirrored expression of the endogenous gene, in that expression required activation both with ionomycin and PMA, was inhibited by cyclosporin A, and was not observed in U937 or THP-1 cells. The region between -108 and -40 bp in the IFN-gamma promoter was required for promoter function and contained two elements that are conserved across species. Deletion of 10 bp within either element reduced promoter function by 70%, whereas deletions in nonconserved portions of this region had little effect on promoter function. The distal conserved element (-96 to -80 bp) contained a consensus GATA motif and a potential regulatory motif found in the promoter regions of the GM-CSF and macrophage inflammatory protein (MIP) genes. Factors binding to this element, including GATA-3, were found in Jurkat nuclear extracts by electromobility shift assays and two of the three complexes observed were altered in response to activation. One or both of these motifs are present in the 5' flank of multiple, other lymphokine genes, including IL-3, IL-4, IL-5, and GM-CSF, but neither is present in the promoter of the IL-2 gene. The proximal conserved element (-73 to 48 bp) shares homology with the NFIL-2A element in the IL-2 promoter; these elements compete for binding of factors in Jurkat nuclear extracts, although the NFIL-2A element but not the IFN-gamma element binds Oct-1. Factors binding to this element in the IFN-gamma gene were present in extracts from resting and activated Jurkat T cells. However, by in vivo footprinting of intact cells, this element was protected from methylation only with activation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8228803 TI - Evidence for two-stage binding by the 175-kD erythrocyte binding antigen of Plasmodium falciparum. AB - Plasmodium falciparum malaria merozoites invade human erythrocytes bearing sialic acid in a multistage process involving the sialic acid-dependent binding of a malaria molecule, the 175-kD erythrocyte binding antigen (EBA-175). We show here that after the initial interaction of EBA-175 with its sialic acid-containing erythrocyte determinant, endogenous proteases can cleave EBA-175 to 65-kD fragment(s), whose binding to erythrocytes is sialic acid independent. A 65-kD fragment was immunoprecipitated by antibodies against peptides between residues 354 and 1061 but not beyond residue 1062. Binding experiments utilizing combinations of native protein, expression-PCR-synthesized EBA-175 polypeptides, peptide synthesis, and antibodies, demonstrated that sialic acid-independent binding could be further mapped to a small (about 40-amino acid) homologous part of the dimorphic allelic region of EBA-175, residues 898-938 (Camp strain numbering). These data support a two-step binding hypothesis and are discussed in relation to the formation of a junction between the merozoite and the erythrocyte, and the finding that after the interaction of some viruses with specific cellular receptors, they undergo conformational changes or cleavage permitting membrane fusion with the host cell. PMID- 8228804 TI - Suppression of B lymphopoiesis during normal pregnancy. AB - We describe a dramatic reduction in numbers and activity of committed B lymphocyte precursors in the bone marrow of normal pregnant mice. Changes in cells responsive to IL-7 were evident as early as 6.5 d of pregnancy and values were < 10% of normal at parturition. B lineage precursors, identified by display of CD45R and absence of surface IgM, were also substantially depressed, and subpopulations representing different stages in the B lineage were assessed by three-color flow cytometry. Early pro-B cells are medium to large in size and have been previously characterized by low expression of the heat-stable antigen (HSA). This category of cells was not reduced, and in fact may have been slightly elevated, during pregnancy. In contrast, all subsequent populations of B lineage precursors, defined by patterns of expression of heat-stable and CD43 antigens, were substantially depressed. The immediate precursors of B cells (small pre-B cells) were identified by small size, expression of CD45R, absence of CD43, and lack of surface IgM. These were the most reduced of any phenotypically defined population in bone marrow. Numbers of newly formed B cells, characterized by the presence of sIgM, but not sIgD, were also diminished. However, B cells with a mature phenotype (sIgM+, sIgD+) were present in normal to somewhat elevated numbers. Mitogen-responsive B cells clonable in a semisolid agar assay were not significantly affected. A bromodeoxyuridine (BrdU) labeling technique was used to evaluate mitotic activity, which revealed an increased proportion of long-lived lymphocytes in the bone marrow of pregnant mice. These observations indicate that B lymphopoiesis is markedly downregulated during pregnancy and that all precursor populations beyond the early pro-B cell stage are affected. The pregnancy-related changes in bone marrow were selective for B lineage precursors, as cells expressing myeloid and erythroid markers were not reduced. In spleen, evidence was obtained for partial depletion of one subset of B cells. These cells, which have been reported to be recent immigrants from marrow, are characterized as having high levels of sIgM and HSA. Changes in other major B lymphocyte subsets in the spleen were less remarkable. When considered with results from the BrdU labeling procedure, the findings indicate that both production and export of lymphocytes from marrow may be substantially decreased. Numbers of B cell precursors were higher in postpartum animals whose litters were removed at birth, suggesting that lactation may prolong regeneration of lymphocyte production.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8228805 TI - p21ras and calcineurin synergize to regulate the nuclear factor of activated T cells. AB - In T lymphocytes, triggering of the T cell receptor (TCR) induces several signaling cascades which ultimately synergize to induce the activity of the nuclear factor of activated T cells (NFAT), a DNA binding complex critical to the inducibility and T cell specificity of the T cell growth factor interleukin 2. One immediate consequence of T cell activation via the TCR is an increase in cytosolic calcium. Calcium signals are important for NFAT induction, and recent studies have identified calcineurin, a calcium-calmodulin dependent serine threonine phosphatase, as a prominent component of the calcium signaling pathway in T cells. A second important molecule in TCR signal transduction is the guanine nucleotide binding protein, p21ras, which is coupled to the TCR by a protein tyrosine kinase dependent mechanism. The experiments presented here show that expression by transfection of mutationally activated calcineurin or activated p21ras alone is insufficient for NFAT transactivation. However, coexpression of the activated calcineurin with activated p21ras could mimic TCR signals in NFAT induction. These data identify calcineurin and p21ras as cooperative partners in T cell activation. PMID- 8228806 TI - Distribution and kinetics of superantigen-induced cytokine gene expression in mouse spleen. AB - The polyclonal stimulation of T cells by bacterial superantigens is involved in the pathogenesis of the toxic shock syndrome in certain staphylococcal and streptococcal infections. Here we describe the onset and kinetics of superantigen induced cytokine production in situ in spleens of normal BALB/c mice monitored at the level of cytokine mRNA expression by in situ hybridization. Messenger RNAs for interleukin 2 (IL-2), interferon gamma, and tumor necrosis factors (TNF) alpha and beta were not expressed at detectable levels in spleens of unstimulated animals but became visible already 30 min after intraperitoneal application of 50 micrograms staphylococcal enterotoxin B. All mRNA levels showed peak expression approximately 3 h after injection and a slow decrease up to 24 h after injection. Expression of the mRNAs was restricted to the T cell-dependent area of the periarteriolar lymphatic sheets of the spleen. Interestingly, TNF-alpha mRNA showed a biphasic response, the early appearing mRNA had the same localization as the other mRNAs, whereas after 3 h TNF-alpha mRNA showed a broader distribution indicating a second cell population producing TNF-alpha. The expression of IL-2 and TNF proteins in the serum increased in parallel to the observed mRNA changes with a slight delay. The presence of macrophages was not required for the expression of the cytokine mRNAs in the spleen as the expression was unchanged in macrophage-depleted mice. Only the second phase of TNF-alpha mRNA expression was abrogated in such animals. The expression of all mRNAs was completely suppressed by prior administration of cyclosporin A. These data show that nonphagocytic cells are the essential superantigen-presenting cells in vivo and indicate that at least part of the pathogenetic TNF-alpha is T cell derived. PMID- 8228807 TI - Mechanisms of class I restricted immunopathology. A transgenic mouse model of fulminant hepatitis. AB - The molecular and cellular mechanisms responsible for cytotoxic T lymphocyte (CTL)-induced immunopathology are not well defined. Using a model in which hepatitis B surface antigen (HBsAg)-specific CTL cause an acute necroinflammatory liver disease in HBsAg transgenic mice, we demonstrate that class I-restricted disease pathogenesis is an orderly, multistep process that involves direct as well as indirect consequences of CTL activation. It begins (step 1) almost immediately as a direct antigen-specific CTL-target cell interaction that triggers the HBsAg-positive hepatocyte to undergo programmed cell death (apoptosis). It progresses (step 2) within hours to a focal inflammatory response in which antigen-nonspecific lymphocytes and neutrophils amplify the local cytopathic effect of the CTL. The most destructive pathogenetic function of the CTL, however, is to secrete interferon gamma when they encounter antigen in vivo, thereby activating the intrahepatic macrophage and inducing a delayed-type hypersensitivity response (step 3) that destroys the liver and kills the mouse. We propose that the principles illustrated in this study are generally applicable to other models of class I-restricted, CTL-induced immunopathology, and we suggest that they contribute to the immunopathogenesis of viral hepatitis during hepatitis B virus infection in humans. PMID- 8228808 TI - Interleukin 5 induces S mu-S gamma 1 DNA rearrangement in B cells activated with dextran-anti-IgD antibodies and interleukin 4: a three component model for Ig class switching. AB - The cellular signals required for induction of immunoglobulin (Ig) class switching are only partially understood. Two processes that are considered to be necessary for such induction are DNA synthesis and germline constant heavy (CH) gene transcription. We now show that an additional signal, as mediated by interleukin 5 (IL-5), is also required. To induce proliferation of resting B cells, but not Ig secretion, we utilized anti-IgD antibodies conjugated to dextran (alpha delta-dex). The addition of IL-4, a well-established switch factor for the IgG1 subclass, to alpha delta-dex-activated cell cultures failed to induce IgG1 secretion or mIgG1+ cells unless IL-5 was also present. While IL-4 stimulated an increase in germline gamma 1 RNA in alpha delta-dex-activated cells, this effect could neither be induced nor enhanced by IL-5. By contrast, IL 5 strongly enhanced steady-state levels of productive gamma 1 RNA induced by alpha delta-dex and IL-4, suggesting that IL-5 stimulated IgG1 switch rearrangement. To test this possibility we measured switch (S) mu-S gamma 1 DNA recombination events using a newly developed assay, digestion circularization polymerase chain reaction (DC-PCR). We demonstrated that IL-5 was necessary for induction of S mu-S gamma 1 DNA rearrangement in alpha delta-dex plus IL-4 activated cells but that it had little effect on rearrangement in the absence of IL-4. Our data strongly suggest, therefore, a three-component model for induction of Ig class switching. This model includes germline CH gene transcription, DNA synthesis, and a third component that is necessary for recombination. PMID- 8228809 TI - Transfer of the inflammatory disease of HLA-B27 transgenic rats by bone marrow engraftment. AB - We have previously produced lines of rats transgenic for HLA-B27 and human beta 2 microglobulin (h beta 2m) that develop a progressive inflammatory disease sharing many clinical and histologic features with the B27-associated human spondyloarthropathies, including gut and male genital inflammation, arthritis, and psoriasiform skin lesions. Other transgenic lines that express lower levels of B27 and h beta 2m remain healthy. To investigate the cellular basis for the multisystem inflammatory disease in these rats, we transferred lymphoid cell populations from disease-prone transgenic lines to irradiated disease-resistant transgenic and nontransgenic recipients. In recipients of cells from two different disease-prone lines, successful transfer required engraftment of bone marrow cells. Transfer of disease with fetal liver cells suggested that neither mature effector cells nor active disease in the donors was necessary for induction of disease in the recipients. Remission of the spontaneous disease in irradiated transgenic rats was induced by engraftment of nontransgenic bone marrow. These results suggest that the expression of HLA-B27 in bone marrow derived cells alone is sufficient for the development of B27-associated disease, and that disease transfer requires engraftment of a bone marrow precursor cell for which mature cells in spleen or in lymph node cannot substitute. PMID- 8228810 TI - Recombinant soluble human Fc gamma RII: production, characterization, and inhibition of the Arthus reaction. AB - A recombinant soluble form of human Fc gamma RII (rsFc gamma RII) was genetically engineered by the insertion of a termination codon 5' of sequences encoding the transmembrane domain of a human Fc gamma RII cDNA. Chinese hamster ovary cells were transfected with the modified cDNA and the secreted rsFc gamma RII purified from the tissue culture supernatant (to > 95%, assessed by SDS-PAGE) using heat aggregated human immunoglobulin G (IgG) immunoaffinity chromatography. The IgG purified rsFc gamma RII was relatively homogeneous (approximately 31,000 M(r)) whereas the total unpurified rsFc gamma RII secreted into the tissue culture supernatant was heterogeneous relating to N-linked glycosylation differences. Functional in vitro activity of the rsFc gamma RII was demonstrated by: (a) ability to bind via the Fc portion of human IgG and mouse IgG (IgG2a > IgG1 > > IgG2b); (b) complete inhibition of binding of erythrocytes sensitized with rabbit IgG to membrane-bound Fc gamma RII on K562 cells; and (c) inhibition of the anti Leu4-induced T cell proliferation assay. Blood clearance and biodistribution studies show the rsFc gamma RII was excreted predominantly through the kidney in a biphasic manner, with an alpha-phase (t1/2 approximately 25 min) and a beta phase (t1/2 approximately 4.6 h); the kidneys were the only organs noted with tissue-specific accumulation. In vivo, the administration of rsFc gamma RII significantly inhibited the immune complex-mediated inflammatory response induced by the reversed passive Arthus reaction model in rats. There was a specific and dose-dependent relationship between the amount of rsFc gamma RII administered, and the reduction in the size and severity of the macroscopic inflammatory lesion. Histological analysis of the skin showed a diffuse neutrophil infiltrate in both control and rsFc gamma RII-treated rats, however the perivascular infiltrate and the red cell extravasation was less intense in the rsFc gamma RII treated group. It is likely that complement activation leads to neutrophil chemotaxis, but neutrophil activation via Fc gamma RII, which results in inflammatory mediator release, is inhibited. The data indicate that rsFc gamma RII is a potential therapeutic agent for the treatment of antibody or immune complex-mediated tissue damage. PMID- 8228811 TI - CD28-induced costimulation of T helper type 2 cells mediated by induction of responsiveness to interleukin 4. AB - Type 1 and type 2 cloned T helper (Th) cells are believed to require different antigen-presenting cell (APC)-derived costimuli for proliferation. In the case of Th1-cloned T cells, CD28 signaling costimulates production of autocrine interleukin 2 (IL-2). Th2 cells produce their autocrine growth factor, IL-4, without costimulation, but require APC-derived costimuli, or IL-1, to respond to IL-4. Here we demonstrate that engagement of CD28 on Th2 cells with anti-CD28 antibody or with APC-associated B7 costimulates Th2 responsiveness to IL-4 but does not affect IL-4 or IL-2 production by Th2 cells. Costimulation of Th2 cells via CD28 appears to involve the induction of IL-1 production by Th2 cells, which acts in an autocrine fashion to induce IL-4 responsiveness. These results suggest that CD28-induced costimulation plays an important role in responses mediated by both types of Th cells. PMID- 8228812 TI - Interferon alpha increases the frequency of interferon gamma-producing human CD4+ T cells. AB - An increased ratio of T helper type 2 (Th2)- vs Th1-like cells contributes to the immune dysregulation in allergic disease situations and in many chronic infections, including AIDS. Th2-type immune responses are characterized by Th cells that produce increased levels of interleukin-4 (IL-4) and decreased levels of interferon gamma (IFN-gamma). The induction of either a Th1- or a Th2-like phenotype may be critically controlled by the antigen-presenting cells and their cytokines, e.g., IFN-alpha. In this study we have determined the frequencies of potential IL-4- and/or IFN-gamma-producing T cells in the peripheral blood of randomly selected healthy individuals, and analyzed whether IFN-alpha controls IL 4 and/or IFN-gamma production. Purified CD4+ or CD8+ T cells were stimulated for 24 h via the T cell receptor/CD3 complex in the presence or absence of IFN-alpha, and single IL-4- and IFN-gamma-secreting cells were detected in enzyme-linked immunospot assays. In the absence of IFN-alpha, CD4 cells produced IFN-gamma at frequencies of 1:50-300, and produced IL-4 at frequencies of 1:110-<1:100,000. Addition of IFN-alpha during the activation of CD4 cells increased the levels of IFN-gamma mRNA. As a consequence, the numbers of IFN-gamma-producing CD4 cells and the amounts of secreted IFN-gamma increased 10-fold. In contrast, IFN-alpha did not increase the frequency of IL-4-secreting CD4 cells. In the absence of IFN alpha, addition of exogenous IL-4 to cultures of CD4 cells suppressed IFN-gamma secretion by 70%. However, in the presence of IFN-alpha, IL-4 did not display any suppressive effect. Compared with CD4 cells, CD8 cells produced IFN-gamma more frequently (1:5-10) but IL-4 less frequently (1:5,300 to < 1:100,000). IFN-alpha did not display any effect on the frequency of either IFN-gamma or IL-4 production by CD8 cells. Taken together the results indicate that IFN-alpha increases the frequency of IFN-gamma-secreting CD4 Th cells and antagonizes the suppressive effect of IL-4 on IFN-gamma production. As a consequence, IFN-alpha may favor the induction and maintenance of Th1-like cells and thereby counteract Th2-driven allergic immune responses. PMID- 8228813 TI - Pim-1 levels determine the size of early B lymphoid compartments in bone marrow. AB - The mouse proto-oncogene Pim-1, which encodes two cytoplasmic serine-threonine specific protein kinases, is frequently activated by proviral insertion in murine leukemia virus-induced hematopoietic tumors. Transgenic mice overexpressing Pim-1 show a low incidence of spontaneous T cell lymphomas, whereas null mutant mice lack an obvious phenotype. We have analyzed the early B lymphoid compartment from both null mutant and E mu-Pim-1 transgenic mice. The level of Pim-1 expression appears to be a determining factor in the ability of these cells to respond to the growth factors interleukin 7 (IL-7) and SF (steel factor). The impaired response in null mutant mice could be rescued by introduction of a functional Pim 1 transgene. Moreover, overexpression of Pim-1 facilitates the derivation of primitive lymphoid cell lines that are dependent on combined stimulation with IL 7 and SF or insulin-like growth factor 1. These results for the first time identify the involvement of Pim-1 in a normal cellular function, as an important regulator of early B lymphopoiesis in mice. PMID- 8228814 TI - Determinant capture as a possible mechanism of protection afforded by major histocompatibility complex class II molecules in autoimmune disease. AB - How peptide-major histocompatibility complex (MHC) class II complexes are naturally generated is still unknown, but accumulating evidence suggests that unfolding proteins or long peptides can become bound to class II molecules at the dominant determinant before proteolytic cleavage. We have compared the immunogenicity of hen egg-white lysozyme (HEL) in nonobese diabetic (NOD), (NOD x BALB/c)F1, and E(d) alpha transgenic NOD mice. We find that a response to the subdominant ANOD-restricted determinant disappears upon introduction of an E(d) molecule, and is restored when scission of HEL separates this determinant from its adjoining, competitively dominant, E(d)-restricted determinant. This suggests that the E(d) molecule binds and protects its dominant determinant on a long peptide while captured neighboring determinants are lost during proteolysis. These results provide clear evidence for "determinant capture" as a mechanism of determinant selection during antigen processing and a possible explanation for MHC-protective effects in insulin-dependent diabetes mellitus. PMID- 8228815 TI - Molecular regulation of the human IL-3 gene: inducible T cell-restricted expression requires intact AP-1 and Elf-1 nuclear protein binding sites. AB - Interleukin 3 (IL-3) is a hematopoietic stem-cell growth and differentiation factor that is expressed solely in activated T and NK cells. Studies to date have identified elements 5' to the IL-3 coding sequences that regulate its transcription, but the sequences that confer T cell-specific expression remain to be clearly defined. We have now identified DNA sequences that are required for T cell-restricted IL-3 gene transcription. A series of transient transfections performed with human IL-3-chloramphenicol acetyltransferase (CAT) reporter plasmids in T and non-T cells revealed that a plasmid containing 319 bp of 5' flanking sequences was active exclusively in T cells. Deletion analysis revealed that T cell specificity was conferred by a 49-bp fragment (bp -319 to -270) that included a potential binding site for AP-1 transcription factors 6 bp upstream of a binding site for Elf-1, a member of the Ets family of transcription factors. DNaseI footprint and electrophoretic mobility shift assay analyses performed with MLA-144 T cell nuclear extracts demonstrated that this 49-bp region contains a nuclear protein binding region that includes consensus AP-1 and Elf-1 binding sites. In addition, extracts prepared from purified human T cells contained proteins that bound to synthetic oligonucleotides corresponding to the AP-1 and Elf-1 binding sites. In vitro-transcribed and -translated Elf-1 protein bound specifically to the Elf-1 site, and Elf-1 antisera competed and super shifted nuclear protein complexes present in MLA-144 nuclear extracts. Moreover, addition of anti-Jun family antiserum in electrophoretic mobility shift assay reactions completely blocked formation of the AP-1-related complexes. Transient transfection studies in MLA-144 T cells revealed that constructs containing mutations in the AP-1 site almost completely abolished CAT activity while mutation of the Elf-1 site or the NF-IL-3 site, a previously described nuclear protein binding site (bp. -155 to -148) in the IL-3 promoter, reduced CAT activity to < 25% of the activity given by wild-type constructs. We conclude that expression of the human IL-3 gene requires the AP-1 and Elf-1 binding sites; however, unlike other previously characterized cytokine genes such as IL-2, the AP-1 and Elf-1 factors can bind independently in the IL-3 gene.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8228816 TI - Protease inhibitors selectively block T cell receptor-triggered programmed cell death in a murine T cell hybridoma and activated peripheral T cells. AB - The hypothesis that cytoplasmic proteases play a functional role in programmed cell death was tested by examining the effect of protease inhibitors on the T cell receptor-mediated death of the 2B4 murine T cell hybridoma and activated T cells. The cysteine protease inhibitors trans-epoxysuccininyl-L-leucylamido-(4 guanidino) butane (E-64) and leupeptin, the calpain selective inhibitor acetyl leucyl-leucyl-normethional, and the serine protease inhibitors diisopropyl fluorophosphate and phenylmethylsulfonyl fluoride, all showed dose-dependent blocking of the 2B4 death response triggered by the T cell receptor complex and by anti-Thy-1. These protease inhibitors enhanced rather than inhibited IL-2 secretion triggered by T cell receptor cross-linking, showing that they did not act by preventing signal transduction. Growth inhibition induced by cross-linking the 2B4 T cell receptor, measured by inhibition of thymidine incorporation, was not generally blocked by these protease inhibitors. All five of these protease inhibitors enhanced rather than blocked 2B4 cell death triggered by dexamethasone, an agent previously shown to have a death pathway antagonistic with that of the TCR. 2B4 cytolysis by the cytotoxic agents staphylococcal alpha toxin and dodecyl imidazole, and that caused by hypotonic conditions, was not significantly affected by the five protease inhibitors tested. The selected protease inhibitors blocked both the apoptotic nuclear morphology changes and DNA fragmentation induced by T cell receptor cross-linking, and enhanced both these properties induced by dexamethasone in 2B4 cells. The T cell receptor-induced death of activated murine lymph node T cells and human peripheral blood CD4+ T cells was blocked by both cysteine and serine protease inhibitors, showing that the protease-dependent death pathway also operates in these systems. PMID- 8228817 TI - Regulation of T cell receptor expression in immature CD4+CD8+ thymocytes by p56lck tyrosine kinase: basis for differential signaling by CD4 and CD8 in immature thymocytes expressing both coreceptor molecules. AB - Signals transduced through the T cell antigen receptor (TCR) are modulated by the src family tyrosine kinase p56lck (lck), which associates in mature T cells with the coreceptor molecules CD4 and CD8. Here we describe a novel function of lck in immature CD4+CD8+ thymocytes, that of regulating TCR expression. Activation of lck in immature CD4+CD8+ thymocytes by intrathymic engagement of CD4 maintains low TCR expression by causing most TCR components to be retained and degraded within the endoplasmic reticulum. Importantly, activation of lck in immature CD4+CD8+ thymocytes results from engagement of surface CD4 molecules, but not surface CD8 molecules, despite the nearly fourfold greater surface expression of CD8 than CD4. The competence of CD4 to activate lck in CD4+CD8+ thymocytes relates to the fact that a relatively large fraction of surface CD4 molecules (25 50%) are associated with intracellular lck molecules, whereas only 2% of surface CD8 molecules are associated with lck. The amount of lck associated with CD4 in CD4+CD8+ thymocytes is diminished by chronic CD4 engagement in the thymus, as activated lck molecules subsequently dissociate from CD4. Indeed, the amount of lck associated with CD4 in CD4+CD8+ thymocytes is markedly increased in major histocompatibility complex (MHC) class II- mice that lack the intrathymic ligand for CD4 and in which surface CD4 molecules are consequently not engaged. Thus, the present study demonstrates that (a) activation of lck in CD4+CD8+ thymocytes regulates distribution and expression of TCR components; (b) unlike CD4 molecules, CD8 molecules on CD4+CD8+ thymocytes cannot efficiently activate lck despite their significantly greater surface expression; and (c) the amount of lck associated with CD4 in the CD4+CD8+ thymocytes is inversely related to the extent of CD4 engagement by MHC class II molecules in the thymus. PMID- 8228818 TI - Response to influenza infection in mice with a targeted disruption in the interferon gamma gene. AB - Interferon gamma (IFN-gamma) is a pleiotropic cytokine secreted by T lymphocytes and natural killer (NK) cells and has been noted to be a first line of host defense in the control of viral infections. To examine further the role of this cytokine in the control of viral infections, mice with a targeted mutation in the IFN-gamma gene were infected with influenza virus, and the in vivo antibody and cell-mediated immune response to viral infection were examined. In addition, cell lines and clones were derived from the immunized animals and the in vitro cytokine production and cytotoxic T lymphocyte (CTL) response were analyzed. The absence of IFN-gamma led to increased production of influenza-specific IgG1, IL 4, and IL-5 as compared to wild-type littermate control animals. In contrast, there was no difference noted in the development of an effective CTL response between IFN-gamma-deficient and wild-type animals. In this model of experimental influenza infection, IFN-gamma is not necessary for the development of an effective humoral or cellular immune response to challenge with this respiratory virus. PMID- 8228819 TI - Macrophage colony-stimulating factor stimulates survival and chemotactic behavior in isolated osteoclasts. AB - Macrophage colony-stimulating factor (M-CSF) is known to play an important role in osteoclast formation. However, its actions on mature cells have not been fully characterized. We now report that M-CSF dramatically stimulates osteoclastic motility and spreading; osteoclasts responded to a gradient of M-CSF with orientation, and random cell polarization occurred after isotropic exposure. M CSF also supported the survival of osteoclasts by preventing apoptosis. Paradoxically, M-CSF inhibits bone resorption by isolated osteoclasts. We found that this was effected predominantly by reduction in the number of excavations. Thus, M-CSF showed a propensity to suppress resorption through a reduction in the proportion of cells that were resorbing bone. Our data suggest that apart from the established role of M-CSF in the provision of precursors for osteoclastic induction, a major role for M-CSF in bone resorption is to enhance osteoclastic survival, migration, and chemotaxis. It seems appropriate that during these processes resorptive functions should be suppressed. We suggest that M-CSF continues to modulate osteoclastic activity once osteoclasts are on resorptive sites, through regulation of the balance between resorption and migration, such that not only the quantity, but the spatial pattern of resorption can be controlled by adjacent M-CSF-secreting cells of osteoblastic lineage. PMID- 8228820 TI - Immature thymocytes become sensitive to calcium-mediated apoptosis with the onset of CD8, CD4, and the T cell receptor expression: a role for bcl-2? AB - During intrathymic negative selection by clonal deletion, crosslinking of the T cell receptor (TCR) induces cell death by delivering an apoptotic signal(s) to the nucleus along a calcium-dependent pathway. We investigated the reactivity of early precursor-containing thymocytes to Ca(2+)-induced signals, and discovered a breakpoint in their sensitivity to calcium-mediated cell death (CMCD). CD25+CD8-4 TCR- (triple negative [TN]) thymocytes stimulated with a calcium ionophore maintain their viability and precursor activity. By contrast, their immediate progeny, CD25-CD8lo4loTCR alpha beta lo (triple low [TL]) cells react to calcium elevation by abrogation of precursor activity and apoptotic cell death. This developmental difference is specific for CMCD, since both CD25+TN and CD25-TL cells are susceptible to steroid-induced apoptosis. The presence of bcl-2 mRNA correlates directly to the resistance to CMCD-CD25+ TN cells express it and CD25 TL cells do not. These experiments show that thymocytes become sensitive to Ca(2+)-induced apoptosis as soon as they begin to express molecules that mediate thymic selection, and suggest that a concomitant downregulation of bcl-2 may mediate this phenomenon. PMID- 8228821 TI - Antigen-induced B lymphocyte activation involves the p21ras and ras.GAP signaling pathway. AB - Ligation of a B lymphocyte surface immunoglobulin (sIg) antigen receptor (AgR) by its specific Ag ligand initiates a signaling pathway that culminates in B cell activation. However, many events of this pathway have not been elucidated. Here we present three novel findings that demonstrate directly that AgR-mediated signaling in B cells functions by the p21ras/ras.GAP-dependent pathway. First, stimulation of TA3 7.9 Ag-specific murine B lymphoma cells for 2 min with either Ag or F(ab')2 anti-IgM induces p21ras activation as measured by an increase in the GTP/GDP ratio of its bound nucleotides. This activation of p21ras does not occur via a change in its guanine nucleotide exchange rate. Second, Ag stimulation results in the inhibition of activity of p120 ras.GAP, a protein that regulates p21ras activation. Tyrosine phosphorylation of ras.GAP occurs within 1 min after Ag stimulation but is no longer detectable at 20 min after stimulation, at which time ras.GAP activity remains inhibited. Thus, tyrosine phosphorylation of ras.GAP is not required for the inhibition of its activity. Third, despite the role proposed for a ras.GAP-associated p190 protein in the control of ras.GAP activity in B cells, p190 was not detectable either in anti-ras.GAP immunoprecipitates of [35S]methionine labeled lysates of Ag-stimulated or unstimulated 7.9 cells or as a tyrosine phosphoprotein in Western blots of anti ras.GAP immunoprecipitates of Ag-stimulated 7.9 cell lysates. Inasmuch as the TA3 7.9 B lymphoma is representative of a mature, sIgM-bearing B cell, our observations raise the intriguing possibility that the capacity of p190 to associate with ras.GAP and regulate the activities of ras.GAP and p21ras in a B cell is dependent on the stage of differentiation of the B cell. PMID- 8228822 TI - A bacterium lipopolysaccharide that elicits Guillain-Barre syndrome has a GM1 ganglioside-like structure. AB - There is a strong association between Guillain-Barre syndrome (GBS) and Penner's serotype 19 (PEN 19) of Campylobacter jejuni. Sera from patients with GBS after C. jejuni infection have autoantibodies to GM1 ganglioside in the acute phase of the illness. Our previous work has suggested that GBS results from an immune response to cross-reactive antigen between lipopolysaccharide (LPS) of the Gram negative bacterium and membrane components of peripheral nerves. To clarify the pathogenesis of GBS, we have investigated whether GM1-oligosaccharide structure is present in the LPS of C. jejuni (PEN 19) that was isolated from a GBS patient. After extraction of the LPS, the LPS showing the binding activity of cholera toxin, that specifically recognizes the GM1-oligosaccharide was purified by a silica bead column chromatography. Gas-liquid chromatography-mass spectrometric analysis has shown that the purified LPS contained Gal, GalNAc, and NeuAc, which are sugar components of GM1 ganglioside. 1H NMR methods [Carr-Purcell-Meiboom Gill (CPMG), total correlation spectroscopy (TOCSY), and nuclear Overhauser effect spectroscopy (NOESY)] have revealed that the oligosaccharide structure [Gal beta 1-3 GalNAc beta 1-4(NeuAc alpha 2-3)Gal beta] protrude from the LPS core. This terminal structure [Gal beta 1-3GalNAc beta 1-4(NeuAc alpha 2-3)Gal beta] is identical to the terminal tetrasaccharide of the GM1 ganglioside. This is the first study to demonstrate the existence of molecular mimicry between nerve tissue and the infectious agent that elicits GBS. PMID- 8228823 TI - Inhibition of T cell and antibody responses to house dust mite allergen by inhalation of the dominant T cell epitope in naive and sensitized mice. AB - Antigen-specific CD4+ T cells play an important role in the allergic immune response to house dust mite (HDM) allergens in humans. The group 1 allergen of Dermatophagoides spp. is a major target antigen in both B and T cell recognition of HDM. In vitro studies have shown that the presentation of peptides to human T cells under appropriate conditions may lead to a state of specific nonresponsiveness. Therefore, to determine if peptides are able to modulate the function of allergen-reactive T cells in vivo, we have used a murine model of T cell recognition of the HDM allergen Der p 1. The results demonstrate that inhalation of low concentrations of peptide containing the major T cell epitope of Der p 1 (residues 111-139), induces tolerance in naive C57BL/6J mice such that they become profoundly unresponsive to an immunogenic challenge with the intact allergen. When restimulated in vitro with antigen, lymph node T cells isolated from tolerant mice secrete very low levels of interleukin 2, proliferative poorly, and are unable to provide cognate help to stimulate specific antibody production. Furthermore, intranasal peptide therapy was able to inhibit an ongoing immune response to the allergen in mice and this has potential implications in the development of allergen-based immunotherapy. PMID- 8228824 TI - A major costimulatory molecule on antigen-presenting cells, CTLA4 ligand A, is distinct from B7. AB - CTLA4 ligands are important costimulatory molecules because soluble CTLA4Ig blocks the induction of T cell responses and induces T cell tolerance. As CTLA4 immunoglobulin (CTLA4Ig) binds B7 when the latter is expressed on fibroblasts, it was widely assumed that CTLA4Ig blocks T cell costimulation by blocking the function of B7. Here we show that the major costimulatory ligand bound by CTLA4Ig (which we term CTLA4 ligand A) on antigen-presenting cells are not encoded by the B7 gene. CTLA4 ligand A also differs from B7 in cellular distribution and in the respective levels of expression. Both B7 and CTLA4 ligand A are critically involved in T cell costimulation. PMID- 8228825 TI - Efficient loading of identical viral peptide onto class II molecules by antigenized immunoglobulin and influenza virus. AB - Several prior reports have identified peptides that are naturally associated with major histocompatibility complex (MHC) class II molecules on presenting cells. We have examined the delivery of a peptide from exogenous sources to MHC class II molecules. The peptide derives from the influenza virus hemagglutinin (HA) and activates a CD4+ T cell hybridoma. In functional assays of antigen presentation, this epitope is delivered effectively to T cells either in the context of influenza virus or chimeric immunoglobulin (Ig) molecules (Ig-HA) in which the peptide has replaced the CDR3 loop of the heavy chain. We find that the identical 11-mer peptide can be isolated from mouse MHC class II antigens whether the exogenous source of peptide is free HA peptide, the Ig-HA chimera, or ultraviolet inactivated PR8 influenza virus. The Ig-HA chimera proves to be the most efficient vehicle for charging class II molecules via the exogenous route. Given the fact that self Igs represent natural long-lived carriers, we suggest that antigenized Igs have considerable potential for peptide delivery to MHC molecules in situ. PMID- 8228826 TI - Long-term acceptance of major histocompatibility complex mismatched cardiac allografts induced by CTLA4Ig plus donor-specific transfusion. AB - Allograft rejection is a T cell-dependent process. Productive T cell activation by antigen requires antigen engagement of the T cell receptor as well as costimulatory signals delivered through other T cell surface molecules such as CD28. Engagement of CD28 by its natural ligand B7 can be blocked using a soluble recombinant fusion protein, CTLA4Ig. Administration of CTLA4Ig blocks antigen specific immune responses in vitro and in vivo, and we have shown that treatment of rats with a 7-d course of CTLA4Ig at the time of transplantation leads to prolonged survival of cardiac allografts (median 30 d), although most grafts are eventually rejected. Here, we have explored additional strategies employing CTLA4Ig in order to achieve long-term allograft survival. Our data indicate that donor-specific transfusion (DST) plus CTLA4Ig can provide effective antigen specific immunosuppression. When DST is administered at the time of transplantation followed by a single dose of CTLA4Ig 2 d later, all animals had long-term graft survival (> 60 d). These animals had delayed responses to donor type skin transplants, compared with normal rejection responses to third-party skin transplants. Furthermore, donor-matched second cardiac allografts were well tolerated with minimal histologic evidence of rejection. These data indicate that peritransplant use of DST followed by subsequent treatment with CTLA4Ig can induce prolonged, often indefinite, cardiac allograft acceptance. These results may be clinically applicable for cadaveric organ and tissue transplantation in humans. PMID- 8228827 TI - Expression of two alpha chains on the surface of T cells in T cell receptor transgenic mice. AB - CD8+ T cells taken directly from mice expressing a Kb-specific T cell receptor (TCR) transgene expressed the transgenic TCR in a bimodal profile as detected by flow cytometric analysis using a clonotype-specific monoclonal antibody. Those cells expressing the lower density of the transgenic TCR expressed the transgenic beta chain and two different alpha chains on their surface. One alpha chain was the product of the alpha transgene, whereas the other was derived by endogenous rearrangement. This report provides the first demonstration that T cells isolated directly from mice may express two different TCR clonotypes on their surface. The potential consequences of this finding for studies using TCR transgenic mice and for the induction of autoimmunity are discussed. PMID- 8228828 TI - The presence of an endogenous murine leukemia virus sequence correlates with the peripheral expansion of gamma delta T cells bearing the BALB invariant delta (BID) T cell receptor delta. AB - gamma delta T cells participate in immune responses during viral, bacterial, and parasitic infections. However, it is not clear whether they recognize antigens produced by pathogens, or are actually reactive to self-ligands generated during the course of infection. In this paper, we report that the presence of the self ligand that selectively expands a subset of gamma delta T cells correlates with the presence of an endogenous murine leukemia virus (MuLV) in inbred strains of mice. The implications of this observation for gamma delta T cell specificity and function is discussed. PMID- 8228829 TI - Thromboxane A2 receptor is highly expressed in mouse immature thymocytes and mediates DNA fragmentation and apoptosis. AB - We have recently revealed that the thymus is the organ showing the highest expression of thromboxane (TX) A2 receptor in mice. In this study, thymic cell populations expressing the receptor were identified, and the effects of a TXA2 agonist on these cells were examined. Radioligand binding using a TXA2 receptor specific radioligand revealed a single class of binding sites in the thymocytes with an affinity and specificity identical to those reported for the TXA2 receptor. The receptor density in these cells was comparable to that seen in blood platelets. This receptor is most highly expressed in CD4-8- and CD4+8+ immature thymocytes, followed by CD4+8- and CD4-8+ cells. The receptor density in splenic T cells was less than one fifth of that in CD4+8+ cells and no binding activity was detectable in splenic B cells. The addition of a TXA2 agonist, STA2, to thymocytes induced the disappearance of the CD4+8+ cells in a time- and concentration-dependent manner and caused DNA fragmentation. These changes were blocked by a specific TXA2 antagonist, S-145. These results demonstrate that TXA2 induces apoptotic cell death in immature thymocytes by acting on the TXA2 receptor on their cell surface and suggest a role for the TXA2/TXA2 receptor system in the thymic micro-environment. PMID- 8228830 TI - The induction of experimental autoimmune myocarditis in mice lacking CD4 or CD8 molecules [corrected]. AB - Experimental induction of most autoimmune diseases appears to depend on the activation of CD4+ T helper cells, while CD8+ lymphocytes may have a role in disease progression. To study the role of CD4+ and CD8+ T cell subsets in T cell dependent autoimmunity, mice lacking CD4 or CD8 molecules after gene targeting were injected with cardiac myosin to induce organ specific autoimmune myocarditis. Mice homozygous for the CD8 mutation (CD8-/-) developed significantly more severe disease as compared to CD4+/-CD8+/- controls. Surprisingly, CD4-/- mice developed autoimmune myocarditis with infiltration of TCR alpha beta +CD4-CD8- T cells in the heart tissue and appearance of autoantibodies. These data demonstrate that the lack of CD4+ or CD8+ T cells has no significant influence on the initiation of autoimmune myocarditis. CD4+ and CD8+ cells regulate disease severity and these results may explain the occurrence of autoimmunity in CD4 immunodeficiencies. PMID- 8228831 TI - List item memory in rats: effects of delay and delay task. AB - The Serial Position Effect (SPE) was studied in rats using 2 manipulations analogous to those that have been shown to decrease the recency effect but leave the primacy effect intact in human Ss. In Part 1, delays (5 s to 60 s) were imposed between exposure to a sequence of arms presented in a 12-arm radial maze and a subsequent test phase. In Part 2, the effect of free access to food in a short (10-s) delay was examined. The results from Parts 1 and 2 showed that the primacy and recency effects were differentially sensitive to the delay and events within it. In particular the recency effect was found to be more sensitive to disruption from these sources. The present demonstration of a reduction in recency with procedures analogous to those used with humans extends the evidence, suggesting that the SPE obtained in rats and humans is a similar phenomenon. PMID- 8228832 TI - Temporal encoding as a determinant of blocking. AB - A blocking paradigm with rats was used to evaluate whether different temporal information is encoded in simultaneous rather than forward associations. During Phase 1, the blocking conditioned stimulus (CS) was simultaneously or forward paired with an unconditioned stimulus (US). During Phase 2, the pretrained CS occurred in compound with a novel target CS that was paired in a simultaneous or forward manner with the US. Forward pretraining resulted in more blocking of a forward than a simultaneously trained target CS, and simultaneous pretraining resulted in more blocking of a simultaneously than a forward trained target CS. Thus, greater blocking occurred when the blocking and blocked CSs had the same temporal relation to the US. The results support the temporal coding hypothesis and question the necessity of predictive information in blocking. PMID- 8228833 TI - The two-test strategy in the study of inhibitory conditioning. AB - It is commonly believed that both a summation test and a retardation test should be used to determine whether a stimulus becomes inhibitory in consequence of some specified treatment, because the 2 tests together rule out alternative interpretations. Depending, however, on the choice of control treatments, a single test may provide credible evidence of inhibition or both together may not. A comprehensive review of the 2-test literature shows that suitable controls have been used only rarely and that compelling evidence of inhibition is correspondingly rare. The only such evidence now available is provided by retardation tests in experiments with some variation of A+/AB- training as the putatively inhibitory treatment. PMID- 8228834 TI - The stimulus movement effect: allocation of attention or artifact? AB - In previous reports--including one by the author--learning has been shown to benefit by having discriminanda move rather than remain stationary. This stimulus movement effect might be attributed to several theoretical mechanisms, including attention, topological memory, and exposure duration. The series of experiments reported in this article was designed to contrast these potential explanatory factors. Ten rhesus monkeys (Macaca mulatta) were tested on a variety of computerized tasks in which the stimuli remained stationary, flashed, or moved at systematically varied speeds. Performance was significantly best when the sample stimulus moved quickly and was poorest when the stimulus remained stationary. Further analysis of these data and other previously published data revealed that the distribution of the stimulus movement effect across trials supported an attention allocation interpretation. PMID- 8228835 TI - Acquired equivalence between cues trained with a common antecedent. AB - In Experiment 1, rats experienced 2 stimuli (A and N) each preceded by the same event (food) or by different events (food preceded 1 but not the other). N was then paired with shock, and the generalization of conditioned suppression to A was assessed. Generalization was more marked when A and N had been experienced along with a common antecedent. In Experiment 2, 3 stimuli (A, B, and N) were presented in initial training. For 1 group, A and N were preceded by food and B was not; for a 2nd group A alone was preceded by food. In each group, suppression generalized more readily from N to the stimulus that had received the same initial training as had been given to N. Experiment 3 found that generalization was not enhanced between stimuli when 1 had preceded food in initial training and 1 had followed it. These results demonstrate that stimuli that have shared a common antecedent will come to be treated as equivalent. PMID- 8228836 TI - Segregation of spatially superimposed optic flow components. AB - The transparency phenomenon, which arises when a radial and a rotational motion pattern are spatially superimposed, suggests that these motion patterns are processed independently. Indeed, for unrestricted stimulus durations, observers could identify the rotational pattern as clockwise or counterclockwise, and the radial pattern as expansion or contraction, even under uncertainty. However, when the time available to process the compound stimulus was equal to the minimum duration required to identify each of the patterns when presented in isolation, identification was impaired. Whereas for spirallike motion patterns the radial and rotational components could be identified, radial and rotational motion patterns were not processed independently when superimposed. Although radial and rotational transformations could not be identified simultaneously, a coherent optic flow pattern could be segregated from another superimposed optic flow component given definite foreknowledge. PMID- 8228837 TI - Optical specification of time-to-passage: observers' sensitivity to global tau. AB - Despite its general mathematical formulation, most empirical work on the visual perception of tau (defined as a quantity divided by its temporal derivative) has focused on the case of direct approach, with tau defined as image angle/rate of expansion. Empirical investigators tend to generalize image size analyses to off axis approaches. However, this generalization is inappropriate for all but a few classes of objects. After mathematically reestablishing the appropriate optical cues specifying time to passage for noncollision cases, we report a series of studies in which we examined observers' sensitivities to this information in both relative- and absolute-judgment paradigms. In general, we found observers' judgments to be accurate and robust. PMID- 8228838 TI - Visual information about time-to-collision between two objects. AB - In a forced-choice paradigm, human observers' sensitivity to visual information specifying a moving object's future time of arrival at a designated position in the field of view was evaluated. A geometrical analysis demonstrated that information specifying a first-order temporal relationship (i.e., without taking changes in velocity into account) is available in the combination of the relative rate of dilation of the optical contour of the moving object and the relative rate of constriction of the optical gap separating the moving object from the target position. Observers were sensitive to information contained in the relative rate of constriction of the optical gap if no contour dilation component was present and to the combination of information contained in the relative rates of dilation of the optical contour of the moving object and constriction of the optical gap if both were present albeit with a differential weighting of the 2 components. PMID- 8228839 TI - Time-to-contact judgment in the locomotion of adults and preschool children. AB - The hypothesis of Lee (1976)--that approach and deceleration toward a surface can be controlled through the rate of change of the optic variable tau--was examined for natural human locomotion. In Experiment 1, 12 adults were asked to perform locomotor tasks that required running at speed and then decelerating so that either the hand or head made a controlled contact with a door. In Experiment 2, 12 preschool children performed a relay-running task that required similar control. In Experiment 3, 12 children and 12 adults ran with a stick as an extension to their arm length and performed the same task. The results supported Lee's hypothesis for the initial phase of approach, but subjects switched to a separate adjustment phase 2 to 3 arm lengths from the target. Children did not adopt an appropriate tau strategy for collision avoidance and appeared unable to modify their approach strategy to allow for a hand-held stick. PMID- 8228840 TI - Perceiving scene layout through an aperture during visually simulated self motion. AB - This study tested scene perception during depicted self-movement. In Experiment 1, viewers reproduced angular scene configurations, revealed over time by camera motions in depth. Viewers used visible flow to update off-screen locations of landmarks that had been drawn from view and thereby perceived scenes in depth beyond the display's boundaries. Experiments 2 and 3 tested whether offscreen space is perceived as a function of depicted velocity and time and whether information from prior views is used. Viewers predicted when lateral movement would reveal a peripheral landmark whose location was shown in a prior panoramic view. Accurate responses, observed under certain conditions, would result if viewers perceived space according to S = integral of V dt (D. Algom & L. Cohen Raz, 1984, 1987; R. Jagacinski, W. Johnson, & R. Miller, 1983). When required to picture wide spans in space yet to emerge, however, viewers responded as if these spans were compressed. The theoretical implications of the ability to retrieve, transform, and apply information after a discontinuous transition from a prior view are discussed. PMID- 8228841 TI - Primacy of dimensions in color perception. AB - In this study, we used a procedure called selective/divided rotation to investigate the role of dimensions in the perception of color. Ss performed either selective-attention or divided-attention tasks to paired dimensions created from each of 3 orientations of axes in color space: 0 degree, 22.5 degrees, and 45 degrees. We evaluated a Euclidean hypothesis, namely, that speeded classification of interacting dimensions is invariant to rigid rotation of stimulus axes. All experiments obtained evidence against this Euclidean hypothesis. Experiments 1 to 4 showed that selective attention was best at the orientation corresponding to saturation and brightness, suggesting primacy of these dimensions. The results were replicated with the pairs hue-saturation (Experiment 7) and hue-brightness (Experiment 8). We conclude that interacting dimensions can be primary and that dimensional primacy characterizes much of perceptual experience. PMID- 8228842 TI - Processing integral dimensions: the whole view. AB - In the traditional view, integral dimensions are said to be processed as unitary whole and only occasionally analyzed. Converging operations establish that (a) pitch and loudness and (b) hue, saturation, and brightness are true psychological dimensions and yet constitute integral dimensions in just this sense. Recent challenges provided by R. D. Melara, L. E. Marks, and their colleagues are shown to be based on narrow and faulty interpretations of evidence for privileged axes. They are also undermined by strong evidence of the holistic processing of pitch and loudness and of the dimensions of color that emerge within both their own data and the larger literature. The traditional view--including the strong claim that integrality entails mandatory holistic processing--continues to fare very well as an account of a substantial and varied set of findings. PMID- 8228843 TI - Destination compatibility, affordances, and coding rules: a reply to Proctor, Van Zandt, Lu, and Weeks. AB - Proctor, Van Zandt, Lu, and Weeks (1993) argued that the invocation of affordances to explain stimulus-response (S-R) compatibilities in reaction time is not needed because left-right direction compatibility, from the coding rules approach, explains the apparent "destination compatibility" effect of Michaels (1988). In this reply, an experiment demonstrates that destination compatibility can be shown even when contradicted by relative left-right motion. The second half of the article addresses theoretical issues separating and joining these two approaches. It is argued that the domain of the affordance approach in S-R compatibility is the guidance of action by information, whereas the domain of coding rules is S-R incompatibility and noncompatibility, situations in which required responses are not afforded and rules must be invoked. The manipulation of rules can mimic some of the consequences of more fundamental perception-action couplings, but principles of the latter sort are needed. PMID- 8228844 TI - Bidirectional grapheme-phoneme activation in a bimodal detection task. AB - A divided attention paradigm was used to investigate whether graphemes and phonemes can mutually activate or inhibit each other during bimodal processing. In 3 experiments, Dutch subjects reacted to visual and auditory targets in single channel or bimodal stimuli. In some bimodal conditions, the visual and auditory targets were nominally identical or redundant (e.g., visual A and auditory /a/); in others they were not (e.g., visual U and auditory /a/). Temporal aspects of cross-modal activation were examined by varying the stimulus onset asynchrony of visual and auditory stimuli. Cross-modal facilitation--but not inhibition- occurred rapidly and automatically between phoneme and grapheme representations. Implications for current models of bimodal processing and word recognition are discussed. PMID- 8228845 TI - Masked partial-word priming in visual word recognition: effects of positional letter frequency. AB - In 4 primed lexical decision experiments, it was found that the positional frequencies of letters shared by the prime and target relative to the overall positional letter frequency of the target determined the magnitude of priming effects. The lower the positional frequency of shared letters, the stronger the facilitatory effect observed. Both an interactive and noninteractive semistochastic version of the interactive activation model captured the principal trends in the data. It is argued that masked partial-word priming arises from a tradeoff between the facilitation generated by prime-target letter overlap and the inhibition generated from all lexical representations activated by letters in the prime that receive further support on target presentation. PMID- 8228846 TI - Multiple bottlenecks in overlapping task performance. AB - A dual-bottleneck model for overlapping-task performance is proposed. A central bottleneck prevents central processes in overlapping tasks from operating in parallel. A response-initiation bottleneck prevents 2 responses from being initiated in close succession. Response requirements of the overlapping tasks were manipulated in order to demonstrate the existence of the latter bottleneck and to assess its role in delimiting overlapping-task performance. In several experiments, elimination of this bottleneck resulted in a strong reduction of underadditive interactions that serve as key indicators of parallel processing in overlapping-task performance. Results indicate that these interactions were caused primarily by the response initiation bottleneck instead of the central bottleneck and suggest that perceptual identification and categorization require the central bottleneck. PMID- 8228847 TI - Response time and accuracy revisited: converging support for the interactive race model. AB - The interactive race model embodies 2 central claims: that divided attention is best described as a race between separately processed codes and that the 2 types of design contingency to which the model is sensitive affect different processing stages. Previous support for the model has come from a series of redundant-target tasks examining reaction time (RT) (J.T. Mordkoff & S. Yantis, 1991). We tested both central claims using near-threshold, accuracy tasks. This approach capitalizes on a known difference between RT and accuracy measures: that (in simple tasks) accuracy is sensitive only to perceptual manipulations, whereas RT is affected by both perceptual and postperceptual factors (J.L. Santee & H.E. Egeth, 1982). The results from 3 experiments provide converging support for the proposed loci of the 2 contingency-sensitive mechanisms within the interactive race model, as well as additional evidence concerning the differential sensitivities of RT and accuracy measures. PMID- 8228848 TI - Orientation-dependent priming effects in the perception of biological motion. AB - In a serial 2-choice reaction time task, subjects discriminated between a biological motion walker and a similar distractor. The point-light walker appeared in 1 of 2 possible in-depth orientations: The figure was walking either to the right or to the left in the sagittal plane. Reliable priming effects were established in consecutive trials but only when priming and primed walkers had the same in-depth orientation. This orientation-dependent priming effect was not tempered when priming and primed figures had different directions of articulatory motion (Experiments 1 to 6), different starting positions in the step cycle (Experiment 2), and different point-light localizations (Experiment 3) or when the figures were translating (Experiments 4 to 6). The data converge with neurophysiological findings that suggest that object recognition is accomplished by accessing high-level, orientation-dependent representations. PMID- 8228849 TI - Insomnia. PMID- 8228850 TI - Mediplan Act. PMID- 8228851 TI - Cervicography. PMID- 8228852 TI - Patient expectations. PMID- 8228853 TI - Endometriosis. PMID- 8228854 TI - Primary care physicians' views on access and health care reform: the situation in North Carolina. AB - BACKGROUND: This cross-sectional study assessed physicians' satisfaction with the current insurance-based reimbursement system and preferences for the two most frequently discussed health care reform proposals, and estimated the association between demographic and practice characteristics and attitudes toward health policy issues and reform plans. METHODS: A random sample of 300 physicians was drawn from state licensure files of general practitioners, family physicians, and pediatricians practicing in North Carolina. All sample physicians were sent a schematic outline of the two major health reform alternatives and a 1-page self administered questionnaire to determine their attitudes toward the current health care system and their preferences for health reform alternatives. RESULTS: Sixty nine percent of physicians responded to the survey. The responses indicated dissatisfaction with the current system and strong beliefs that access to care is inadequate in this diverse state with a large poor and rural population. Nearly one third of the physicians reported having insufficient information to choose between plans. Among physicians expressing a preference, 37% preferred managed competition, 38% preferred continuing the current system, and 25% preferred a single-payer system. CONCLUSIONS: A uniform opinion about health care policy is a thing of the past for American medicine. Because terms used in the health reform debate (especially "managed competition") are ambiguous and set in the context of an increasingly diverse medical profession, no single direction of health reform (much less a specific plan) secures widespread understanding or support from a large proportion of physicians. None of the plans will please all of the doctors all of the time. PMID- 8228855 TI - Physicians' strategies for safeguarding confidentiality: the influence of community and practice characteristics. AB - BACKGROUND: Confidentiality is an issue of particular importance in rural communities where it seems everyone knows everyone else. Privacy may be compromised when professional relationships are interwoven with personal ones. We conducted a study to determine the influence of practice characteristics on safeguarding confidentiality. METHODS: A survey was mailed to all family and general practice physicians in Kansas. Differences in confidentiality strategies were studied to determine if they were related to characteristics of the respondents and their practice environments. RESULTS: A total of 510 physicians (55%) responded to the survey. Only in practices in which more than 5% of the patients were personal acquaintances was a strategy for preserving confidentiality affected: of seven specific strategies, only one, speaking with office personnel about confidentiality, was statistically associated with this practice characteristic. CONCLUSIONS: Physicians practicing in smaller communities and areas with more extensive social interconnectedness may need to address the issue of privileged communication to a greater extent. If other options are exhausted, physicians may need to send patients with particular needs for confidentiality outside their communities to obtain care. PMID- 8228856 TI - A comparison of labor and delivery management between nurse midwives and family physicians. AB - BACKGROUND: Practice associations between family physicians and nurse midwives have been suggested as a means to increase the availability of obstetric care in rural areas. No evidence exists, however, that family physicians and midwives have comparable practice styles or achieve similar outcomes in obstetric patients. METHODS: The study examines patients cared for by a co-practice of nurse midwives and family physicians at a rural hospital. Data were collected through a retrospective chart audit for all patients whose prenatal care, labor, or delivery was managed by members of the practice in 1990 and 1991. RESULTS: Few differences were noted between nurse midwives and family physicians in the management of labor or delivery. The only consistent finding was that family physicians were more likely than midwives to use an episiotomy for delivery (40% vs 30% in primiparous women, P = .02; and 20% vs 10% in multiparous women, P = .007). Despite seemingly similar management styles, primiparous women managed by family physicians were more likely to undergo cesarean section (14% vs 8%, P = .05) resulting from the diagnosis of dystocia. When practice specialty was included in a logistic regression model with parity and the number of preexisting risk factors, the effect of specialty on cesarean sections remained significant with a relative risk of 2.79 for cesarean section if patients had their labor managed by a family physician (P < .001). CONCLUSIONS: Family physicians and nurse midwives managed patients in labor similarly, but nurse midwives were more likely to achieve a vaginal delivery in primiparous women and do so without an episiotomy. Although the differences found would not interfere with a collaborative practice, subtle differences in patient management do exist. Further exploration of these differences may be helpful in understanding the impact of these differences on mixed-specialty practices. PMID- 8228857 TI - A comparison of family physicians' and obstetricians' intrapartum management of low-risk pregnancies. AB - BACKGROUND: We hypothesized that family physicians' style of intrapartum management was less interventional than the management style of obstetricians, and that this would not adversely affect maternal or neonatal outcomes. METHODS: A retrospective, matched-pair study design was used to compare low-risk women cared for by community family physicians with those cared for by obstetricians at a small teaching hospital. The subjects were matched on the basis of age and parity. We compared the rates of intervention between family physicians and obstetricians. RESULTS: We studied 351 matched pairs of women. The demographic characteristics of patients were similar as were the rates for most labor and delivery procedures. Family physicians had lower rates for induction, external and internal fetal monitoring, narcotic analgesia use, and postpartum oxytocin use. Women cared for by family physicians spent less time in the hospital, both during labor and postpartum. CONCLUSION: This study supports the hypothesis that at our center family physicians intervene less than obstetricians in intrapartum management. Comparisons with similar studies conducted at other academic centers illustrate differences in styles of practice between institutions, not just between specialties. PMID- 8228858 TI - Cervicography: an intermediate triage test for the evaluation of cervical atypia. AB - BACKGROUND: A substantial number of abnormal Papanicolaou (Pap) smears are reported as demonstrating "cytologic atypia." This finding may actually represent premalignant cervical disease. Some of these patients may not be able to afford definitive colposcopic examinations, and simply repeating cytologic testing may result in missed treatable disease. The purpose of this study was to evaluate the use of cervicography as an intermediate triage test for women with atypical cervical cytology. METHODS: Women with a recent smear demonstrating cytologic atypia were evaluated using colposcopy, biopsy, and cervicography. RESULTS: Colposcopically directed cervical biopsies were obtained from 224 of 685 women with cytologic atypia. The histologic specimens confirmed evidence of cervical dysplasia for 166 women. Of these women, cervicography detected 74.7% of those who had mild dysplasia, 87.5% of those who had moderate dysplasia, 75% of those who had severe dysplasia, and the one patient who had cervical cancer. Most (93%) of the women with dysplasia that was undetected by cervicography had mild dysplasia. CONCLUSIONS: Cervicography may be an effective intermediate triage test for the evaluation of young women with Pap smears demonstrating cytologic atypia. PMID- 8228859 TI - Physicians' predictions of elderly outpatients' preferences for life-sustaining treatment. AB - BACKGROUND: Research has shown that physicians are poor predictors of patients' life-sustaining treatment preferences. Our study examined the association between three aspects of physician experience and their ability to accurately predict patients' preferences for two different life-sustaining treatments in the event of two serious medical conditions. METHODS: Seventeen physicians predicted the treatment preferences of 57 patients and then interviewed patients regarding their actual treatment preferences. Physicians' professional experience, length of their relationship with the patient, and experience with direct feedback were measured to determine the association of these factors with the accuracy of the physicians' predictions. RESULTS: Physicians became more accurate predictors as they interviewed more patients and received direct feedback regarding the accuracy of their predictions (P < .001). Residents were more accurate than faculty in predicting patients' preferences (P < .05). CONCLUSIONS: Increased experience with life-sustaining treatment discussions improved the physicians' abilities to accurately predict patient preferences. Although possibly resulting from small sample size, neither greater professional experience nor longer relationship with a patient improved the accuracy of physicians' predictions. Future research should examine whether discussing end-of-life issues with patients more often makes physicians more sensitive predictors of patients' life sustaining treatment preferences. PMID- 8228860 TI - A five-minute psychiatric screening interview. AB - Recent studies indicate that as many as 30% of primary care patients have some type of psychiatric disorder. Although common, these disorders are often neither detected nor treated by primary care physicians. A brief interview is described that can be used as an effective screening tool to identify patients who require further evaluation. PMID- 8228861 TI - CD-ROM: a primer for physicians. AB - Compact disks with read-only memory (CD-ROM) are a powerful storage medium and are rapidly becoming standard equipment on personal computers. Each disk has the capacity to store 680 million bytes (megabytes or MB) of text, graphics, photographic images, audio, animation, and full-motion video. Many medical reference texts are currently available in CD-ROM format, with the advantage of improved search, retrieval, cross-referencing, and printing capabilities. PMID- 8228862 TI - Prurigo nodularis. AB - Prurigo nodularis is a chronic skin disorder that is frequently misdiagnosed. It is characterized by multiple pea-sized nodules that develop on the skin, particularly on the anterior leg and thigh. These lesions are intensely pruritic and frequently become excoriated and infected. The cause of prurigo nodularis is unknown, and treatment is directed at reducing inflammation and controlling symptoms. The condition is typically of long duration and is refractory to treatment. PMID- 8228863 TI - Association of penicillin allergy with idiopathic anaphylaxis. AB - Idiopathic urticaria is a common and usually self-limited illness that is occasionally associated with life-threatening idiopathic anaphylaxis. A case is presented in which urticaria existed for 30 years before tests that revealed penicillin allergy as the probable cause. Our findings suggest that evaluation for penicillin allergy may be useful in some cases of urticaria. PMID- 8228864 TI - Preventing substance abuse. An interview paradigm. AB - In summary, substance use and abuse is a major health problem in the United States affecting adolescents of both sexes and of every socioeconomic level. Family physicians have numerous opportunities to improve the health of future generations by using specific skills and expertise to prevent, identify, and intervene with adolescents' abuse of alcohol and other chemical substances. The first step is taking an effective alcohol and drug-use history. PMID- 8228865 TI - Infection with the avian polyomavirus, BFDV, selectively affects myofibril structure in embryonic chick ventricle cardiomyocytes. AB - Embryonic cardiomyocytes can both beat and divide. They assemble cardiac muscle specific proteins into sarcomeric myofibrils and contract. In addition, they periodically synthesize DNA, complete mitosis, disassemble sarcomeric myofibrils in the area of the mitotic spindle, assemble cytoplasmic isoform-specific proteins into a cleavage furrow contractile ring, undergo cytokinesis, and then reform sarcomeric myofibrils in daughter cells. Little is known about how embryonic cardiomyocytes disassemble their myofibrils as they traverse the cell cycle and divide. In the present study, beating embryonic avian ventricular cardiomyocytes in primary culture were stimulated to initiate DNA synthesis without subsequent mitosis or cytokinesis by infection with the lytic avian polyomavirus, Budgerigar Fledgling Disease Virus (BFDV). Within 48 hours, infected, adherent cardiomyocytes disassemble most of their sarcomeric myofibrils, retaining cardiac myosin only in thin myofibrils with disrupted sarcomeric periodicity and in amorphous nonfibrillar pools. By 72 hours, infected cardiomyocytes contain no myofibrils and no longer react with antibodies to cardiac myosin. In contrast, infected cardiomyocytes continue to display cytoplasmic myosin localized in stress-fiber-like-structures in adherent cells, or in disrupted fibers and dispersed pools in detaching cells. Infected cardiomyocytes also continue to display interphase-like arrays of polymerized microtubules, even when rounded-up just prior to lysis. These results suggest that polyomavirus infection may provide a useful model system for further study of the regulation of myofibrils disassembly in embryonic cardiomyocytes. PMID- 8228866 TI - Steroidogenic shift by cultured ovarian follicles of Rana dybowskii at breeding season. AB - The steroid secretory activity of cultured ovarian follicles of Rana dybowskii and the activities of relevant steroidogenic enzymes were examined during the natural hibernation period (October-February). Enzyme activities were measured indirectly by monitoring the conversion of exogenous substrates to products by isolated follicles. Follicles were incubated for 6 h in amphibian Ringer in the presence or absence of frog pituitary homogenate (FPH, 0.1 pituitary/2 ml) and/or various steroid precursors. Progesterone (P4), 17 alpha-hydroxyprogesterone (17 alpha-OHP) or testosterone (T) secreted by the follicles into the medium were measured by RIA. In the presence of FPH, high levels of P4 were produced by follicles at the early and mid-hibernation period (695 and 898 pg/follicle, respectively) whereas markedly elevated levels of P4 were produced during late hibernation (1,393 pg/follicle) (just prior to or breeding season, February). In contrast, high levels of T were produced by the follicles early in hibernation (1,206 pg/follicle) while negligible levels were produced in late hibernation (69 pg/follicle). Higher levels of 17 alpha-OHP were produced by follicles at early and mid hibernation (594 and 705 pg/follicle, respectively) than in later hibernation (221 pg/follicle). Addition of exogenous pregnenolone markedly increased P4 levels in a dose-dependent manner when added to follicles at early and late stages of hibernation: FPH addition further enhanced conversion of pregnenolone. Similarly, addition of androstenedione (AD) increased T levels in a dose-dependent manner by these follicles. However, following addition of exogenous P4, less 17 alpha-OHP was produced by follicles collected in late hibernation as compared to those collected earlier.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228867 TI - Effects of an antiandrogen treatment on morphological characters and physiological functions of male fallow deer (Dama dama L.). AB - From mid-April to late November, i.e., during antler growth phase and main breeding season, a group of four (later three) male fallow deer was treated with increasing doses of the antiandrogen cyproterone acetate (CA). Throughout the treatment period, plasma testosterone levels of the bucks were either undetectable or only slightly above the detection limit of 0.1 ng/ml. In marked contrast to controls, testicular volumes and neck girths of the CA treated bucks were basal during the application period, proving the strong antigonadotropic effect of CA (testicular volume) and the absence of androgen action at the normal target organs (neck girths), respectively. Antler growth in the CA treated bucks occurred during the normal time span for fallow deer and the antlers produced were of species specific shape. From this it is concluded that in fallow deer, onset, duration, and termination of antler growth are independent of androgen action. Velvet shedding in the CA treated bucks however did not occur at the normal time (late August/early September), but was postponed until late December/early January, i.e., about 5 weeks after the termination of CA application. Activity of alkaline phosphatase in the experimental animals increased during the antler growth phase, but was markedly lower than in controls. Maximum body weight in the CA treated bucks was reached already in June, i.e., about 3 months prior to controls. Furthermore, the rut associated, drastic decline in body weight observed in controls did not occur in the experimental animals. PMID- 8228868 TI - Energy metabolism of the trophectoderm and inner cell mass of the mouse blastocyst. AB - The two cell populations of the day 5 mouse blastocyst--the inner cell mass (ICM) and trophectoderm (TE)--were isolated by immunosurgery and TE biopsy. The uptake of glucose and pyruvate and the production of lactate were measured in the isolated cell populations and in single day 5 blastocysts, using a non-invasive ultramicrofluorescence technique. The number of cells in isolated ICMs, TE vesicles, and intact blastocysts were counted after differential labeling with fluorochromes. The uptake of glucose and the formation of lactate were approximately three and five times greater, respectively, in the ICM cells compared with the TE cells. All the glucose consumed by the ICM could be accounted for by lactate formation, whereas the value for TE was 55%. The uptake of pyruvate was low in both cell types. The results are consistent with the proposition that the TE acts as a transporting epithelium, sparing nutrients for metabolism by the ICM. PMID- 8228869 TI - Influence of the mucin coat on the survival of rabbit and mouse embryos stored at 0 degrees C. AB - To examine the factors affecting the survival of refrigerated embryos, rabbit and mouse morulae were stored at 0 degrees C in modified phosphate-buffered saline (PB1) or in PB1 containing 0.75 M sucrose. Survival was defined as the ability to develop into an expanded blastocyst in culture. The data was analyzed with special reference to the presence of a mucin coat around the embryos. When rabbit morulae were stored in isotonic PB1 for 2, 4, 6, and 8 days, survival rates were 98%, 88%, 85%, and 50%, respectively. However, if the mucin coat had been removed before storage, the rates were lower (95%, 75%, 36%, and 3%, respectively). Sucrose impaired the survival of rabbit morulae irrespective of the presence of the mucin coat. Only 11% of mouse morulae survived 2 days of storage in PB1 medium, but if the medium contained sucrose, survival rates after storage for 2, 3, 4, and 5 days were higher (83%, 55%, 31%, and 7%, respectively). To provide them with a mucin coat around the zona pellucida, mouse embryos were incubated in a rabbit oviduct. Survival rates of these embryos after storage in the presence of sucrose did not decrease over 4 days of refrigeration (98-92%), and the rates after storage for 5, 6, and 7 days were 65%, 40%, and 30%, respectively; embryos that had been stored for 5 days were transferred to recipient mice, and live young were born. Agar embedding of mouse morulae did not have the same effect as the mucin coat.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228870 TI - Initiation of the human sperm acrosome reaction by thapsigargin. AB - Previous studies have established that the mammalian sperm acrosome reaction (AR) is dependent upon an influx of extracellular Ca2+, but the involvement of a mobilizable store of intracellular Ca2+ has not been shown. In many other cells, the endoplasmic reticulum is the site of such a Ca(2+)-store. Here, we show that thapsigargin, a highly specific inhibitor of the endoplasmic reticulum Ca(2+) ATPase Ca(2+)-pump (and thus a mobilizer of intracellular Ca2+) in other cells, can initiate the AR in capacitated human sperm. Thapsigargin at concentrations from 50-500 nM significantly increased the AR to the same extent when incubated with capacitated sperm for 1 min (assayed by indirect immunofluorescence). Transmission electron microscopy confirmed the occurrence of normal morphology in the AR initiated by thapsigargin. Thapsigargin (200 nM) did not initiate the AR in noncapacitated sperm. Initiation of the AR by thapsigargin apparently requires an influx of Ca2+ since 1 min preincubation with the calcium channel blockers La3+ (250 microM) or Ni2+ (250 microM) prior to addition of thapsigargin completely inhibits AR-initiation. Mobilization of an intracellular Ca(2+)-store by thapsigargin in capacitated human sperm may lead to an influx of extracellular Ca2+ and subsequently the AR. Putative sites for thapsigargin-sensitive intracellular Ca(2+)-stores in human sperm include the cytoplasmic droplet, the sperm nucleus and the acrosome. PMID- 8228871 TI - Identifying "does" with help of dentures. PMID- 8228872 TI - Invasive Hemophilus influenzae type B disease. PMID- 8228873 TI - The distribution of ethanol in postmortem blood samples. PMID- 8228874 TI - The effects of specific latent fingerprint and questioned document examinations on the amplification and typing of the HLA DQ alpha gene region in forensic casework. AB - The apparent stability of DNA in forensic samples has permitted the successful application of several techniques such as polymerase chain reaction (PCR)-based and restriction fragment length polymorphisms (RFLP) analysis to forensic cases. PCR-based typing of the HLA-DQ alpha region in forensic casework has been shown to be a valid and reliable technique. This inherent stability of DNA in forensic evidence has led us to address the question of whether DNA could successfully withstand certain evidence processes such as latent fingerprint and electrostatic detection apparatus (ESDA) processing and still yield a sufficient quantity and quality of DNA for PCR HLA DQ alpha typing. Through testing done with biological material on simulated and casework envelope, stamp, and cigarette butt type evidence, it was determined that samples could be processed for specific latent fingerprint and ESDA examinations and still yield sufficient DNA for conclusive HLA DQ alpha typing results. PMID- 8228875 TI - A comparison of the fixed bin method with the floating bin and direct count methods: effect of VNTR profile frequency estimation and reference population. AB - When the results of a forensic comparison of highly polymorphic variable number tandem repeat (VNTR) loci fail to exclude a suspect as a possible contributor of biological evidence, it is desirable to convey to the trier of fact the significance of the match. Furthermore, in a forensic context, it is desirable that the estimated frequency of occurrence be conservative, that is, that any uncertainty in the estimate will favor the accused. Using an empirical approach with a data base of 2046 individuals belonging to one of four population groups, this study examined the effect of the method used to estimate frequency of occurrence of a VNTR profile from a reference data base, and the consequences of using a data base that may not represent the circumstances of the crime. The fixed bin method was at least as conservative as the floating bin and genotype counting (direct counting) methods. Secondly, for forensic purposes, profile frequency estimates from different reference populations do not deviate greatly. VNTR profiles are rare in any of the data bases. PMID- 8228876 TI - Deoxyribonuclease I (DNase I) typing from semen stains: low enzyme activity in vaginal fluids does not interfere with seminal DNase I typing from mixture stains. AB - We describe the use of deoxyribonuclease I (DNase I) polymorphism for individualization of semen in body fluid stain mixtures, as a means of providing new and more useful information to practicing forensic biologists as a genetic marker. We have already reported that human DNase I isozyme patterns from different subjects are classifiable into ten groups. Isoelectric focusing of DNase I isozymes on polyacrylamide gel (IEF-PAGE, pH 3.5 to 5) was accomplished using a 0.5 mm thick gel. Pretreatment of semen samples with neuraminidase enhanced the isozyme band resolution and sensitivity. Activity detection using the dried agarose film overlay (DAFO) procedure was reliable, sensitive and simple, with high resolution, and the phenotypes of DNase I were determined in semen stains of about 0.3 microL stored at room temperature for up to a year in most of the samples tested. The DNase I types in semen stains were correlated with the types found in the corresponding blood and urine samples, although most of the vaginal fluid samples had no typable DNase I activity. This is considerably advantageous for seminal individualization from body fluid mixture stains in criminal cases. An evaluation of DNase I typing by IEF-PAGE and DAFO was also performed on casework samples submitted to our laboratory, and the results showed that DNase I was expected to be one of the most useful individualization marker of semen in practical application. PMID- 8228877 TI - Transferrin subtyping in dental pulps. AB - Serum transferrin (TF) subtypes were also found in dental pulps by isoelectric focusing and immunoblotting. The types observed in dental pulps completely agreed with those in serum samples from the same individuals. The allele frequencies in 105 samples were TF*C1 = 0.757 and TF*C2 = 0.243. Reliable subtyping was possible for 4 weeks following extraction of the teeth. The TF system can provide a useful genetic marker for the medicolegal individualization of teeth. PMID- 8228878 TI - Correlates of sex offender and offense traits by victim age. AB - The authors studied offender, offense, and victim characteristics according to victim age cohort among a sample of over 800 incarcerated sex offenders. Their findings were somewhat different from those previously reported, and suggest that rapists of adults were more psychopathic, sexual assailants against adolescents appeared to be more typical family men, and child molesters were more opportunistic chronic sex offenders. The authors believe their findings begin to shed light on the nature of sex crimes committed against victims of different age. PMID- 8228879 TI - Seroprevalence of parenterally transmitted viruses (HIV-1, HBV, HCV, and HTLV I/II) in forensic autopsy cases. AB - HIV, HBV, HCV, and HTLV show similar modes of transmission and infection is frequently identified in certain population groups. The seroprevalence of these infectious agents and interrelation in forensic autopsy populations are described here. A total of 414 serum samples were collected sequentially from autopsy cases at the Office of the Chief Medical Examiner for the State of Maryland. All samples were tested for the presence of antibodies to HIV-1, HCV, HTLV-I/II, and hepatitis B core antigen, using Enzyme-linked Immunosorbent Assays (ELISA). Samples yielding repeatedly reactive results were confirmed by Western Blots (WB) for HIV-1 and HTLV-I/II, while for HCV confirmation, the second generation recombinant immunoblot (RIBA-2) was used. Of the 414 cases, 32.6% (135/414) were infected with at least one of the four viruses. 5.6% were seropositive for HIV-1, 23.2% for HBV, 19.1% for HCV, and 1.0% for HTLV-I/II. Intravenous drug users (IVDU) showed significant increased prevalences of HIV-1, HBV, and HCV. 83.6% of the IVDU were infected with at least one of the three viruses, 25.5% with HIV-1, and 47.3% with both HBV and HCV. The data indicate that the overall prevalence of HIV, HBV, and HCV seropositivity in the autopsy population of an inner city medical examiner office is much higher than that of the general population. Routine testing only for HIV-1 would have missed 86% of the infections with HCV or HBV. Universal precautions should be strictly employed by all personnel working in this environment. PMID- 8228880 TI - Sudden out-of-hospital coronary death in patients with no previous cardiac history. An analysis of 221 patients studied at autopsy. AB - Many epidemiological, clinical, and autopsy studies have demonstrated that the risk of sudden coronary death (SCD) is linked to the severity of coronary atherosclerotic lesions and to their thrombotic potential. However it remains unknown why some individuals manifest no clinical evidence of coronary disease until the onset of sudden death. The aim of this autopsy study has been to analyze the sociodemographic, clinical, and pathological features in case of sudden out-of-hospital coronary death where the death was the first manifestation of coronary disease. The results have been compared with those obtained from victims of SCD with known cardiac history. A total of 377 autopsies of sudden out of-hospital cardiovascular deaths were performed at the Forensic Institute of Paris between 1989 and 1991. A total of 221 were SCD. A total of 160 of these subjects (72.4%) had no cardiac history (group A). The other 61 victims of SCD had cardiac antecedents (group B). Men account for 63% and 77% of the SCD in groups A and B respectively and are 12 years younger than women at the onset of SCD in both groups (65/77 years). Our study showed that in both groups SCD occurs in 83% of cases at home (and at rest) and in 30% of these cases while sleeping. Autopsy showed that SCD be it with or without antecedent occurs on a background of severe coronary disease with multivessels stenoses but coronary thrombosis was rarely observed (15%). SCD occurred in the context of underlying cardiomegaly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228881 TI - Blows to the maternal abdomen causing fetal demise: report of three cases and a review of the literature. AB - Traumatic abruption results in 6% of third-trimester gravidas who are hit or kicked in the abdomen during assaults. Three cases are presented in which blows inflicted to the abdomen of pregnant women in their third trimester resulted in the death of the fetus due to abruptio placentae. Two cases were domestic altercations while one was a third-party criminal assault. In all cases the gravida herself escaped significant intra-abdominal injury, and external abdominal findings were minimal. The clinical signs were a history of loss of fetal movements shortly after the assault and loss of fetal heart tones within hours after the assault. One patient had vaginal bleeding; one had uterine contractions. In the cases of domestic abuse, both women initially gave false histories of how the injury occurred. PMID- 8228882 TI - Neck injuries: III. Ligamentous injuries of the craniocervical articulation without occipito-atlantal or atlanto-axial facet dislocation. A pathologic study of 21 traffic fatalities. AB - Craniocervical ligamentous injuries without dislocation of the occipito-atlantal or atlantoaxial motion segments have been described rarely if at all. Among 155 traffic fatalities were 21 persons with such injuries. Among the 21 fatalities were 20 alar ligament injuries, eight tectorial membrane injuries, and ten injuries of the subaxial cervical spine. All 21 had craniocerebral trauma, including twelve with skull fractures, seven with facial or mandibular fractures, eight with subdural hemorrhages, twelve with subarachnoid hemorrhages, seven with cerebral contusions, five with cerebral lacerations, two with midbrain lacerations, nine with pontomedullary lacerations, and four with spinomedullary injuries. The mechanisms of death was acute neurogenic shock in seven victims, acute neurogenic shock in combination with other physiological derangements in six, acute neurogenic respiratory arrest in three, brain swelling in two, and mechanisms unrelated to head and neck trauma in three victims. In six victims, the neck injury did not contribute to death. No conclusions regarding the biomechanical mode of injury are drawn. PMID- 8228883 TI - Study of the extraction of LSD from illicit blotters for HPLC determination. AB - The effect of different parameters (extraction method, temperature, time, solvent) on the extraction efficiency of LSD from impregnated papers has been investigated. Designed experiments have been applied according to the Plackett Burman method. For the quantitative determination of LSD reversed phase ion-pair chromatography has been used with UV detection. The experimental conditions assuring maximal LSD recovery have been determined, as well as the constant and proportional bias of the extraction procedure elaborated. PMID- 8228884 TI - Microbial DNA challenge studies of variable number tandem repeat (VNTR) probes used for DNA profiling analysis. AB - DNA probes commonly used for forensic DNA profiling analysis were hybridized to Hinfl digested DNA isolated from various common microbial species. Extended exposures to light sensitive film failed to detect any DNA fragments of a microbial origin following hybridizations to radio-labeled DNA insert single locus probes (SLPs) and nonisotopically labeled oligonucleotide SLPs. PMID- 8228885 TI - Multigenerational amplification of a reference ladder for alleles at locus D1S80. AB - The interlaboratory typing of DNA specimens that have been amplified at the D1S80 locus necessitates the use of a standard allelic reference ladder. This communication describes a technique in which individual, amplified alleles are isolated, combined, and amplified by PCR to produce a functional reference ladder composed of many of the alleles that occur at this locus. The amplified ladder can serve directly as a template source for production of the next generation of reference ladder. This process, in which each amplified ladder serves as the template for the next has been carried through multiple generations. PMID- 8228886 TI - A simplified method for phenotyping alpha-2-HS-glycoprotein. AB - Alpha-2-HS-glycoprotein is considered a reliable genetic marker for paternity testing and forensic investigation. We describe a simple, semiautomated method for phenotyping this protein in serum samples, bloodstains and dental pulp using isoelectric focusing methods followed by immunofixation techniques. This method produces clear results even with very old biological evidence. PMID- 8228887 TI - Comparison of postmortem autolysis in cardiac and skeletal muscle. AB - To understand the mechanism in postmortem autolysis better, processes in the postmortem degradation of myofibril proteins in the presence of protease inhibitors were studied. Male Wistar rats were given injections of the carboxyl-, thiol-, and serine-protease inhibitors, pepstatin, Ep-475[L-transepoxysuccinyl leucylamide(3-methyl) butane; E-64-C], and chymostatin, via the femoral vein. Control rats were similarly treated with saline. Then, myofibril proteins were isolated from their cardiac and femoral muscles and from those of control animals at various times after death, and degradation of these myofibril proteins with time was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In cardiac muscle, alpha-actinin was degraded rapidly, followed by the heavy chain of myosin and light chain of myosin (L2). Actin and the light chain of myosin (L1) were degraded slowly. the degradations of the heavy chain of myosin, alpha-actinin, tropomyosin and L2 after 14 days were not inhibited by pepstatin, but were inhibited by Ep-475 and chymostatin. In skeletal muscle, L1 and L2 were degraded rapidly, followed by the heavy chain of myosin and alpha-actinin. Actin was degraded slowly and was still unchanged 2 weeks after death. The degradations of protein components were inhibited by pepstatin, Ep-475 and chymostatin. These results indicated that after death the components of myofibrils are degraded by various proteases at various rates depending on their properties or structures. This degradation is fundamentally the same in cardiac and skeletal muscles, but inhibitors have somewhat different effects on the postmortem degradation processes after death in the two types of muscle. PMID- 8228888 TI - A systematic approach to the sampling of dental DNA. AB - As investigations into the forensic aspects of DNA analysis continue, the human tooth will play a dual role in identification. Dentin and enamel provide a protective enclosure for genomic and mitochondrial DNA as well as providing the basis for radiographic, biochemical, and ultrastructural forensic studies. The purpose of this investigation is to establish technical guidelines, based on histology and experimental evidence, for the management and sampling of dental DNA. The anatomic location of dental DNA is discussed with emphasis on the conservation of tooth structure during sampling. Ten pairs of maxillary right and left third molars were sampled for DNA following storage for 18 weeks at ambient temperature and humidity. Right third molars were crushed, whereas the left third molars were sectioned conservatively prior to sampling the DNA. The quantity and quality of human DNA obtained from each tooth was compared, as well as the radiographic appearance of remaining hard tissue and the overall simplicity of each approach. DNA typing was performed, both sequence and length based analyses, comparing teeth from the same individual and teeth from different donors. The results of this study suggest that the odontologist will maximize the dental DNA yield by crushing the entire specimen but that substantial yields of human DNA can be obtained by using a conservative technique that preserves the tooth structure. In addition, the method of sampling does not affect the ability to perform DNA typing analyses. PMID- 8228889 TI - GC/MS quantitation of benzoylecgonine following liquid-liquid extraction of urine. AB - An analytical procedure for the determination of benzoylecgonine in urine by GC/MS using a two step liquid-liquid extraction scheme has been developed. Benzoylecgonine is one of the principle metabolites of cocaine. The extraction uses a wash step prior to extraction and derivatization and requires 0.5 mL of urine. Benzoylecgonine is extracted in good % recovery at a basic pH by using 4 mL of dichloromethane. The method is accurate and sensitive and allows the quantitation of benzoylecgonine in urine at or below the National Institute on Drug Abuse confirmation cutoff level of 150 ng/mL. PMID- 8228890 TI - Endomyocardial fibrosis in a scuba diving death. AB - A death of an amateur SCUBA diver with undiagnosed late phase endomyocardial fibrosis is reported. This 27-year-old man found himself in difficulties during a Sub-Aqua Club outing and drowned. Autopsy revealed features of drowning in addition to severe endomyocardial fibrosis (EMF) with collagenization of the left ventricle and interventricular septum with foci of chronic myocarditis. Other organs were essentially normal and toxicological studies were negative. Examination of the air tank revealed a normal oxygen circulation with elevation of the carbon dioxide content above recommended limits. Sudden ventricular arrhythmia underwater due to EMF is believed to have precipitated drowning. Stricter medical monitoring of amateur divers should prevent similar accidents in the future. PMID- 8228891 TI - The incredible bouncing bullet: projectile exit through the entrance wound. AB - A case of a suicidal gunshot wound of the head where the projectile ricocheted within the calvarium and exited through the entrance wound is reported. Findings at the scene and from examination of the victim and the suspect weapon are presented and discussed. PMID- 8228892 TI - A condom--the critical link in a rape. AB - An intact condom, reputedly used during a rape, was submitted for forensic examination. Conventional biochemistry results indicated that blood found on one side of the condom may have originated from the victim. Semen from the other side of the condom was not characterizable by conventional biochemical methods. Pubic hairs recovered from the condom matched those of the victim and not those of the suspect. Testing the blood and semen from the condom by DNA analysis gave the profile of the victim from the blood and the profile of the suspect from the semen. PMID- 8228893 TI - Road to PRO contract fraught with twists and turns. PMID- 8228894 TI - Health-care reform will also re-form academic medical centers. PMID- 8228895 TI - Clinical diagnosis. Brilliant and not so. PMID- 8228896 TI - Politics and medicine. Inseparable and dependent. PMID- 8228897 TI - Dr. Jacob Ernst Brecht. Missionary, physician and friend to the Seminoles. PMID- 8228898 TI - Anesthesiology in the forgotten war. PMID- 8228899 TI - Lincoln's last physician. Lesson from history for today. PMID- 8228900 TI - Politics, poison and plague. Mariel boat lift legacy. PMID- 8228901 TI - Yellow fever and hookworm disease. Impetus for public health in the southern United States. PMID- 8228902 TI - Polymerase chain reaction from bench to bedside. Applications for infectious disease. AB - The past decade has seen the rapid advancement of molecular biology and its application in the field of infectious diseases. The polymerase chain reaction (PCR) is a technique which allows for the in vitro amplification of DNA. The ability of this method to recognize and amplify small quantities of foreign genome from unpurified samples will revolutionize the diagnoses of microbial diseases which have previously been cumbersome or impossible. It has already been widely applied in this area, and clinical labs will soon be offering this test for selected pathogens. There are ongoing studies evaluating its use in CNS infections and for the identification of viruses in the immunocompromised host. The PCR is not without problems, particularly false positive results from contamination. Because it is so sensitive, the significance of a positive result in patients with latent or chronic infections is yet to be determined. As our knowledge and familiarity with PCR expands, it will become a powerful tool for the clinician to use to identify various infectious diseases. PMID- 8228903 TI - Should physicians have a voice in insurer settlements of medical malpractice claims? PMID- 8228904 TI - Funds for physician families in crisis. PMID- 8228905 TI - Malpractice attorneys will be our heroes. PMID- 8228906 TI - Significant changes in doctor-patient relationship. PMID- 8228907 TI - cAMP-activated apical membrane chloride channels in Necturus gallbladder epithelium. Conductance, selectivity, and block. AB - Elevation of intracellular cAMP levels in Necturus gallbladder epithelium (NGB) induces an apical membrane Cl- conductance (GaCl). Its characteristics (i.e., magnitude, anion selectivity, and block) were studied with intracellular microelectrode techniques. Under control conditions, the apical membrane conductance (Ga) was 0.17 mS.cm-2, primarily ascribable to GaK. With elevation of cell cAMP to maximum levels, Ga increased to 6.7 mS.cm-2 and became anion selective, with the permeability sequence SCN- > NO3- > I- > Br- > Cl- >> SO4(2-) approximately gluconate approximately cyclamate. GaCl was not affected by the putative Cl- channel blockers Cu2+, DIDS, DNDS, DPC, furosemide, IAA-94, MK-196, NPPB, SITS, verapamil, and glibenclamide. To characterize the cAMP-activated Cl- channels, patch-clamp studies were conducted on the apical membrane of enzyme treated gallbladders or on dissociated cells from tissues exposed to both theophylline and forskolin. Two kinds of Cl- channels were found. With approximately 100 mM Cl- in both bath and pipette, the most frequent channel had a linear current-voltage relationship with a slope conductance of approximately 10 pS. The less frequent channel was outward rectifying with slope conductances of approximately 10 and 20 pS at -40 and 40 mV, respectively. The Cl- channels colocalized with apical maxi-K+ channels in 70% of the patches. The open probability (Po) of both kinds of Cl- channels was variable from patch to patch (0.3 on average) and insensitive to [Ca2+], membrane voltage, and pH. The channel density (approximately 0.3/patch) was one to two orders of magnitude less than that required to account for GaCl. However, addition of 250 U/ml protein kinase A plus 1 mM ATP to the cytosolic side of excised patches increased the density of the linear 10-pS Cl- channels more than 10-fold to four per patch and the mean Po to 0.5, close to expectations from GaCl. The permeability sequence and blocker insensitivity of the PKA-activated channels were identical to those of the apical membrane. These data strongly suggest that 10-pS Cl- channels are responsible for the cAMP-induced increase in apical membrane conductance of NGB epithelium. PMID- 8228908 TI - Two types of sugar-binding protein in the labellum of the fly. Putative taste receptor molecules for sweetness. AB - Flies have taste cells specifically sensitive to sweetness. It has been suggested that the cells possess two types of receptor sites covering the receptive field of sweetness. By affinity electrophoresis with the site-specific inhibitory polysaccharides, two types of sugar-binding protein were isolated from the labellar extract of the blowfly. These proteins showed consistent sugar-binding specificities and affinities with the two types of receptor sites for sweetness, respectively. The dissociation constant of the protein-sugar complex varies 100 400 mM and the molecular weight of one type of the protein is 27,000, while that of the other is 31,000 or 32,000. Both proteins were water insoluble and were also detected in the isolated chemosensilla. Thus they are probably located on the taste receptor membrane, and the proteins are likely to act as the taste receptor molecules for sweetness in the fly. PMID- 8228909 TI - Voltage-gated transient currents in bovine adrenal fasciculata cells. I. T-type Ca2+ current. AB - The whole cell version of the patch clamp technique was used to identify and characterize voltage-gated Ca2+ channels in enzymatically dissociated bovine adrenal zona fasciculata (AZF) cells. The great majority of cells (84 of 86) expressed only low voltage-activated, rapidly inactivating Ca2+ current with properties of T-type Ca2+ current described in other cells. Voltage-dependent activation of this current was fit by a Boltzmann function raised to an integer power of 4 with a midpoint at -17 mV. Independent estimates of the single channel gating charge obtained from the activation curve and using the "limiting logarithmic potential sensitivity" were 8.1 and 6.8 elementary charges, respectively. Inactivation was a steep function of voltage with a v1/2 of -49.9 mV and a slope factor K of 3.73 mV. The expression of a single Ca2+ channel subtype by AZF cells allowed the voltage-dependent gating and kinetic properties of T current to be studied over a wide range of potentials. Analysis of the gating kinetics of this Ca2+ current indicate that T channel activation, inactivation, deactivation (closing), and reactivation (recovery from inactivation) each include voltage-independent transitions that become rate limiting at extreme voltages. Ca2+ current activated with voltage-dependent sigmoidal kinetics that were described by an m4 model. The activation time constant varied exponentially at test potentials between -30 and +10 mV, approaching a voltage-independent minimum of 1.6 ms. The inactivation time constant (tau i) also decreased exponentially to a minimum of 18.3 ms at potentials positive to 0 mV. T channel closing (deactivation) was faster at more negative voltages; the deactivation time constant (tau d) decreased from 8.14 +/- 0.7 to 0.48 +/- 0.1 ms at potentials between -40 and -150 mV. T channels inactivated by depolarization returned to the closed state along pathways that included two voltage-dependent time constants. tau rec-s ranged from 8.11 to 4.80 s when the recovery potential was varied from -50 to -90 mV, while tau rec-f decreased from 1.01 to 0.372 s. At potentials negative to -70 mV, both time constants approached minimum values. The low voltage-activated Ca2+ current in AZF cells was blocked by the T channel selective antagonist Ni2+ with an IC50 of 20 microM. At similar concentrations, Ni2+ also blocked cortisol secretion stimulated by adrenocorticotropic hormone. Our results indicate that bovine AZF cells are distinctive among secretory cells in expressing primarily or exclusively T-type Ca2+ channels.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8228910 TI - Voltage-gated transient currents in bovine adrenal fasciculata cells. II. A-type K+ current. AB - In whole cell patch clamp recordings on enzymatically dissociated adrenal zona fasciculata (AZF) cells, a rapidly inactivating A-type K+ current was observed in each of more than 150 cells. Activation of IA was steeply voltage dependent and could be described by a Boltzmann function raised to an integer power of 4, with a midpoint of -28.3 mV. Using the "limiting logarithmic potential sensitivity," the single channel gating charge was estimated to be 7.2 e. Voltage-dependent inactivation could also be described by a Boltzmann function with a midpoint of 58.7 mV and a slope factor of 5.92 mV. Gating kinetics of IA included both voltage-dependent and -independent transitions in pathways between closed, open, and inactivated states. IA activated with voltage-dependent sigmoidal kinetics that could be fit with an n4h formalism. The activation time constant, tau a, reached a voltage-independent minimum at potentials positive to 0 mV. IA currents inactivated with two time constants that were voltage independent at potentials ranging from -30 to +45 mV. At +20 mV, tau i(fast) and tau i(slow) were 13.16 +/- 0.64 and 62.26 +/- 5.35 ms (n = 34), respectively. In some cells, IA inactivation kinetics slowed dramatically after many minutes of whole cell recording. Once activated by depolarization, IA channels returned to the closed state along pathways with two voltage-dependent time constants which were 0.208 s, tau rec-f and 10.02 s, tau rec-s at -80 mV. Approximately 90% of IA current recovered with slow kinetics at potentials between -60 and -100 mV. IA was blocked by 4 aminopyridine (IC50 = 629 microM) through a mechanism that was strongly promoted by channel activation. Divalent and trivalent cations including Ni2+ and La3+ also blocked IA with IC50's of 467 and 26.4 microM, respectively. With respect to biophysical properties and pharmacology, IA in AZF cells resembles to some extent transient K+ currents in neurons and muscle, where they function to regulate action potential frequency and duration. The function of this prominent current in steroid hormone secretion by endocrine cells that may not generate action potentials is not yet clear. PMID- 8228911 TI - The transduction system in the isoproterenol activation of the Ca(2+)-activated K+ channel in guinea pig taenia coli myocytes. AB - In freshly dispersed guinea pig taenia coli myocytes the activity of the large conductance Ca(2+)-activated K+ channel (maxi-K+ channel) predominates. The open probability (Po) of this channel is increased by micromolar concentrations of the beta-adrenergic agonist isoproterenol (ISO). Low concentrations of cholera toxin (CTX, 1 pM) and guanosine 5'-O-2-thiodiphosphate (GDP beta S, 0.5 mM) suppress the ISO-induced increase of Po. Higher concentrations of CTX (e.g., 0.5 nM) as well as forskolin and dibutyryl cAMP increase the Po. 1,9-Dideoxyforskolin, the forskolin analogue, which lacks the adenylate cyclase-stimulating effect, does not. A specific protein kinase A inhibitor (Wiptide), applied intracellularly via diffusion from the patch electrode, suppresses the ISO-induced increase of whole cell outward K+ current during step depolarization. In contrast, intracellularly applied protein kinase C (19-36), a specific protein kinase C inhibitor, has no effect on the whole-cell current. TMB-8, an inhibitor of intracellular calcium mobilization, does not affect either the whole-cell outward K+ current during step depolarization or the Po. These observations show that ISO increases the Po of the maxi-K+ channels in the guinea pig taenia coli myocytes through the G protein-adenylate cyclase-protein kinase A system. PMID- 8228912 TI - Dopamine modulates in a differential fashion T- and L-type calcium currents in bass retinal horizontal cells. AB - White bass (Roccus chrysops) retinal horizontal cells possess two types of voltage-activated calcium currents which have recently been characterized with regard to their voltage dependence and pharmacology (Sullivan, J., and E. M. Lasater. 1992. Journal of General Physiology. 99:85-107). A low voltage-activated transient current was identified which resembles the T-type calcium current described in a number of other preparations, along with a sustained high threshold, long-lasting calcium current that resembles the L-type calcium current. Here we report on the modulation of horizontal cell calcium channels by dopamine. Under whole-cell voltage clamp conditions favoring the expression of both calcium currents, dopamine had opposing actions on the two types of voltage sensitive calcium currents in the same cone-type horizontal cell. The L-type calcium current was significantly potentiated by dopamine while the T-type current was simultaneously reduced. Dopamine had no effect on calcium currents in rod-type horizontal cells. Both of dopamine's actions were mimicked with the D1 receptor agonist, SKF 38393, and blocked by application of the D1 specific antagonist, SCH 23390. Dopamine's actions on the two types of calcium currents in white bass horizontal cells are mimicked by the cell membrane-permeant cyclic AMP derivative, 8-(4-chlorophenylthio)-cyclic AMP, suggesting that dopamine's action is linked to a cAMP-mediated second messenger system. Furthermore, the inhibitor of cAMP-dependent protein kinase blocked both of dopamine's actions on the voltage-dependent calcium channels when introduced through the patch pipette. This indicates that protein phosphorylation is involved in modulating horizontal cell calcium channels by dopamine. Taken together, these results show that dopamine has differential effects on the voltage-dependent calcium currents in retinal horizontal cells. The modulation of these currents may play a role in shaping the response properties of horizontal cells. PMID- 8228913 TI - Effect of fura-2 on action potential-stimulated calcium release in cut twitch fibers from frog muscle. AB - Cut fibers (striation spacing, 3.6-4.2 microns) were mounted in a double Vaseline gap chamber and studied at 14-15 degrees C. One or both of the Ca indicators fura 2 and purpurate-3,3' diacetic acid (PDAA) were introduced into the optical recording site by diffusion from the end pools. Sarcoplasmic reticulum (SR) Ca release was elicited by action potential stimulation. With resting [fura-2] = 0 mM at the optical site, the [Ca] transient measured with PDAA was used to estimate SR Ca release (Baylor, S.M., W.K. Chandler, and M.W. Marshall. 1983. Journal of Physiology. 344:625-666). With resting [fura-2] > 0 mM, the contribution from Ca complexation by fura-2 was added to the estimate. When resting [fura-2] was increased from 0 to 0.5-2 mM, both the amount of SR Ca release and the maximal rate of release were increased by approximately 20%. These results are qualitatively similar to those obtained in intact fibers (Baylor, S.M., and S. Hollingworth. 1988. Journal of Physiology. 403:151-192; Hollingworth, S., A. B. Harkins, N. Kurebayashi, M. Konishi, and S. M. Baylor. 1992. Biophysical Journal. 63:224-234) and are consistent with a reduction of Ca inactivation of SR Ca release produced by 0.5-2 mM fura-2. With resting [fura-2] > or = 2 mM, the PDAA [Ca] transient was reduced to nearly zero and SR Ca release could be estimated from delta [Cafura-2] alone. When resting [fura-2] was increased from 2-4 to 5-6 mM, both the amount of SR Ca release and the maximal rate of release were decreased by approximately half, consistent with a possible reduction of Ca-induced Ca release (Jacquemond, V., L. Csernoch, M. G. Klein, and M. F. Schneider. 1991. Biophysical Journal. 60:867-873) or a possible pharmacological effect of fura-2. PMID- 8228914 TI - Reduction of calcium inactivation of sarcoplasmic reticulum calcium release by fura-2 in voltage-clamped cut twitch fibers from frog muscle. AB - Cut fibers from Rana temporaria and Rana pipiens (striation spacing, 3.9-4.2 microns) were mounted in a double Vaseline-gap chamber and studied at 14 degrees C. The Ca indicator purpurate-3,3' diacetic acid (PDAA) was introduced into the end pools and allowed to diffuse into the optical recording site. When the concentration at the site exceeded 2 mM, step depolarizations to 10 mV were applied and the [Ca] transient measured with PDAA was used to estimate Ca release from the sarcoplasmic reticulum (SR) (Baylor, S. M., W. K. Chandler, and M. W. Marshall. 1983. Journal of Physiology. 344:625-666). With depolarization, the rate of SR Ca release increased to an early peak and then rapidly decreased several-fold to a quasi-steady level. The total amount of Ca released from the SR at the time of peak rate of release appeared to be independent of SR Ca content, consistent with the idea that a single activated channel might pass, on average, a fixed number of ions, independent of the magnitude of the single channel flux. A possible explanation of this property is given in terms of locally induced Ca inactivation of Ca release. The solution in the end pools was then changed to one with PDAA plus fura-2. SR Ca release was estimated from the [Ca] transient, as before, and from the delta [Cafura-2] signal. On average, 2-3 mM fura-2 increased the quasi-steady level of the rate of SR Ca release by factors of 6.6 and 3.8, respectively, in three fibers from Rana temporaria and three fibers from Rana pipiens. The peak rate of release was increased in five of the six fibers but to a lesser extent than the quasi-steady level. In all fibers, the amplitude of the free [Ca] transient was markedly reduced. These increases in the rate of SR Ca release are consistent with the idea that Ca inactivation of Ca release develops during a step depolarization to 10 mV and that 2-3 mM fura-2 is able to reduce this inactivation by complexing Ca and thereby reducing free [Ca]. Once the concentration of fura-2 becomes sufficiently large, a further increase reduces the rate of SR Ca release. On average, 5-6 mM fura-2 increased the quasi-steady rate of release, compared with 0 mM fura-2, by 6.5 and 2.9, respectively, in four fibers from Rana temporaria and three from Rana pipiens.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8228915 TI - Absence of human papillomavirus type 16 E6 transcripts in HPV 16-infected, cytologically normal cervical scrapings. AB - By a combination of reversed transcription and subsequent polymerase chain reaction (RNA-PCR), 23 cytologically normal cervical scrapings, positive for the presence of human papillomavirus type 16 (HPV 16) DNA, were analyzed for the presence of transcripts originating from the E6 region of the viral genome. This region is thought to be involved in transformational, tumorigenic events. No mRNAs of the E6 region were detectable using the most sensitive PCR-mediated procedure currently available. Since it was previously shown that in cytological abnormal cervical scrapings about one-half of the samples positive for HPV 16 DNA express mRNAs of the E6 region, a difference between normal and abnormal cervical scrapings, when the HPV 16 is present, exists. The observed difference between cytologically normal and abnormal, HPV-DNA-positive cervical scrapes may eventually be used as a prognostic marker for screening of women at risk for the development of cervical carcinoma. However, firm establishment of the putative correlation between tumor progression and the presence of E6 transcripts requires extensive follow-up analysis of HPV-positive patients. PMID- 8228916 TI - Hepatitis C virus infection in hemodialysis patients in southern Sweden: epidemiological, clinical, and diagnostic aspects. AB - A prevalence of hepatitis C virus (HCV) antibodies of 12% was found in 276 patients from 11 dialysis units. Between zero and 22% of the patients in the different units were anti-HCV positive. The epidemiology of HCV was studied in two units during a 2 year period by antibody assays and the polymerase chain reaction and correlated with clinical manifestations. Two types of epidemiologic patterns were found that may explain the wide difference of HCV prevalence described in different dialysis units. In one unit there was no evidence of spread within the unit, and the prevalence of HCV was dependent on the status of the patients entering for treatment. In the other unit, a clustering of infected patients could be seen in which 13 of 36 were infected during a 3 year period. Some patients who had not received blood transfusions were among the infected. Hepatitis C infection was the most common explanation for repeated abnormal transferase levels. Most of the HCV-infected patients reacted both for anti-HCV and HCV RNA. HCV RNA was in general detected earlier than anti-HCV seroconversion. Among 20 HCV RNA-positive serum samples that were anti-HCV ELISA positive 18 had indeterminate and two negative reactions by immunoblot (RIBA 2). Thus the RIBA 2 test should be used with caution as a confirmatory antibody test in this group of patients. PMID- 8228917 TI - Diagnosis of enteroviral meningitis with IgG-EIA using heat-treated virions and synthetic peptides as antigens. AB - Two recently developed enzyme immunosorbent assays (EIA) for the detection of significant titre increases in enteroviral IgG-antibodies were evaluated as diagnostic tools in 127 etiologically well-characterized patients with aseptic meningitis. One assay was based on heat-treated virions (H-EIA) and one on synthetic peptides (P-EIA) as antigens. The sensitivities, with virus isolation as reference method, were 0.67 by H-EIA and 0.62 by P-EIA, which were higher than by a routinely used complement fixation test (CFT, 0.51) but somewhat lower than the sensitivities found by two previously presented IgM-assays, mu-capture EIA, and solid-phase reverse immunosorbent test (SPRIST). The specificities of the two IgG-EIA techniques and CFT were apparently high, whereas the two IgM-assays showed positive reactions in some non-enteroviral cases. A relatively rapid increase in enteroviral IgG-antibodies was apparent using H-EIA and P-EIA. The two IgG-EIA tests contributed with considerable additional etiological information since significant IgG-rises were obtained in 13 patients by H-EIA and in 19 by P-EIA, respectively, out of the 56 individuals in whom enterovirus isolation was negative and a non-enteroviral diagnosis was not found. Thus, detection of enteroviral IgG-antibodies by H-EIA and P-EIA seems to be a valuable alternative to CFT for the routine diagnosis of enteroviral meningitis. The IgM assays, mu-capture EIA, and SPRIST, may allow a relatively rapid report of an enteroviral infection. However, since both the IgM-tests are hampered by incomplete specificities, a confirmation of positive results by an IgG-assay should be carried out. PMID- 8228918 TI - Epstein-Barr virus genomic sequences and specific antibodies in cerebrospinal fluid in children with neurologic complications of acute and reactivated EBV infections. AB - Four children with infectious mononucleosis (IM) and one with reactivated Epstein Barr virus (EBV) infection had concomitant central nervous system disorders. Cerebrospinal fluid (CSF) samples from all five patients contained EBV genomic sequences and EBV-specific antibodies in the neurologic stage, but not during convalescence. Cerebrospinal fluid from two non-neurologic IM patients had neither EBV DNA nor EBV antibodies. The EBV-positive CSF of the five with neurological disorders were aseptic in culture and all negative for other human herpesvirus DNAs and antibodies: herpes simplex virus types 1 and 2, cytomegalovirus, varicella-zoster virus, and human herpesvirus 6. Epstein-Barr virus DNA and EBV antibodies were not detected in the CSF of 17 EBV-seropositive patients with mumps meningitis, rubella encephalitis, unknown febrile convulsion, or partial epilepsy. It is suggested that EBV plays a causal role in neurologic manifestations in patients with acute and reactivated EBV infections, through direct viral invasion and immunopathological reactions. PMID- 8228919 TI - Diagnostic significance of antibodies to hepatitis B virus polymerase in acutely and chronically HBV-infected individuals. AB - The prevalence and time course of the occurrence of antibodies to the hepatitis B virus polymerase (anti-HBpol) were investigated in acutely and in chronically HBV infected individuals by using recombinant HBpol protein for Western blot analysis. One group consisted of 19 patients who were acutely infected and recovered completely. Five of these patients (26%, 69 serum samples examined) exhibited anti-HBpol. Among those anti-HBpol positive patients, recovery from the disease was combined with a complete loss of this antibody. In contrast, in a second group of 15 individuals who developed chronic hepatitis B, 13 (87%, 102 serum samples examined) had anti-HBpol during the acute phase of the disease. The difference between the anti-HBpol prevalence rates of the two patient groups is statistically significant (Exact Fisher test, P < .002), implying that the occurrence of anti-HBpol may be indicative of a potential chronic course of hepatitis B. Remarkably, anti-HBpol was found in one case of a clinically suspected hepatitis B in which no other serological HBV parameters were found. This serum sample was positive in HBV PCR, supporting a possible diagnostic value of anti-HBpol. PMID- 8228920 TI - Genetic variability of German hepatitis C virus isolates. AB - Heterogeneity of hepatitis C viral (HCV) genomes of several isolates from different countries has been reported, but there is little information on HCV isolates for the Federal Republic of Germany. Therefore, the nucleotide (nt) and deduced amino acid (aa) sequences of interesting parts of the viral genome derived from different human isolates in Germany were compared with each other and with the nt and predicted aa sequences of recently published isolates. HCV sequences were obtained by reverse transcription of viral RNA extracted from serum followed by polymerase chain reaction (PCR) amplification. Within the 5' nontranslated region we found only 3 single nucleotide exchanges among 2 of our isolates, and in comparison to sequences of Japanese isolates 2 to 3 exchanges, and to U.S. isolates 1 to 5 exchanges (homologies 98% to > 99%). Determination of a 249-bp core sequence from two German isolates exhibited 3% sequence divergence. The sequence of the core region (nt 342-911) showed a homology of about 88-91% on nt level and 96-97% on aa level as compared to U.S. isolates and other German isolates, and a homology of 95-96% (nt) and 96-98% (aa), respectively, to Japanese isolates. Less homologies were noticed for the E1 and E2/NS1 genes, especially in the N-terminal E2/NS1 hypervariable domain. Our isolates HD1 and HD2 showed nt sequence homologies of about 72-81% and aa homologies of 76-88% to U.S., German, and French isolates, and 89-91% (nt) and 88-96% (aa), respectively, to Japanese isolates. These results indicate that various German isolates are more closely related to Japanese isolates and differ from other European isolates as reported so far. Because of a nucleotide sequence heterogeneity of up to 10% among the tested isolates, we conclude that more than one closely related but distinct viral genotype of HCV exists in Germany. Furthermore, heterogeneous sequences of HCV can be detected in a single patient suggesting multiple infection with different genomic variants or, alternatively, a genetic drift forced by mutational events as a consequence of host immune selection. PMID- 8228921 TI - Quantitative assessment of hepatitis B virus DNA in chronic hepatitis B: comparison of two solution hybridization assays. AB - We compared two solution hybridization assays, the AffiProbe assay (Orion Corporation) and the Abbott HBV-DNA assay (Abbott), for quantitative measurement of hepatitis B virus (HBV) DNA in serum samples obtained from chronic hepatitis B surface antigen (HBsAg) carriers. Forty transversally collected (group 1) and 83 serially collected (group 2) serum samples from chronic hepatitis B patients were tested with both assays. The serial serum samples were obtained from 6 patients who underwent alpha-interferon therapy with different outcomes (nonresponse, hepatitis B e antigen [HBeAg] seroconversion, HBeAg and HBsAg seroconversion). In group 1 a good correlation (r = 0.91; P < 0.001) was found between the HBV-DNA results of the two assays. Two samples (5%) were HBV-DNA positive according to the Abbott but negative according to the AffiProbe assay; for all other samples the HBV-DNA status corresponded. In group 2 the assays gave colinear HBV-DNA results during follow-up of 5 of the 6 patients (correlation for the total group: r = 0.90; P < 0.001). Nevertheless, in both groups the AffiProbe assay yielded about 5-10 times higher HBV-DNA levels than the Abbott HBV-DNA assay (P < 0.001). These discordant results were most probably due to standardization differences of the positive control samples of the two test systems. This observation underlines the need for international standardization of HBV-DNA and uniform reference panels. PMID- 8228922 TI - Prevalence of canine distemper antibodies in the pagetic population. AB - Recent molecular evidence has implicated canine distemper virus (CDV) as a possible aetiologic agent in Paget's disease. However, previous serological studies have shown no differences in levels of anti-CDV antibodies between Paget's patients and controls. In this study, the technique of enzyme linked immunosorbent assay was used to measure anti-CDV antibodies in a group of Paget's patients from the North West of England. Some patients were undergoing treatment with 3-amino-hydroxypropylidene (APD), and the pre-treatment levels of antibody were compared with those following treatment. With several patients, it was also possible to compare the antibody levels with results from in situ hybridisation studies. No significant difference was found between the levels of anti-CDV antibodies in patients and controls. However, several patients and some of the controls did have markedly elevated levels of anti-CDV antibody. Antibody levels remained fairly constant following treatment with APD, except for two patients who showed marked changes. The patients positive for CDV by in situ hybridisation had significantly lower levels of anti-CDV antibodies when compared with those that were negative by in situ hybridisation. These results suggest that if CDV does cause Paget's disease, anti-CDV antibodies play little or no part in the disease pathogenesis. High levels of anti-CDV antibodies in both Paget's patients and controls suggest that a canine virus can infect humans. The fact that those patients that had CDV transcripts detectable in their bone cells had low levels of anti-CDV antibodies might suggest failure to clear the virus during an initial infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228923 TI - Hepatitis C antibodies in dialysis patients and patients with leukaemia. AB - The prevalence of hepatitis C antibodies (anti-HCV) among multitransfused patients was studied and compared with predicted values obtained from a post transfusion hepatitis study and from data on the prevalence of anti-HCV among blood donors. The prevalence of hepatitis B core antibodies (anti-HBc) was also studied to determine the routes of transmission of hepatitis C virus. The patients consisted of 65 dialysis patients (57 on haemodialysis and 8 on continuous ambulatory peritoneal dialysis) and 71 leukaemia patients in long-term remission [49 with acute myeloid leukaemia (AML) and 22 with acute lymphatic leukaemia (ALL)]. The presence of anti-HCV was investigated using a second generation enzyme-linked immunosorbent assay. Reactive samples were confirmed by a second generation recombinant immunoblot assay. Anti-HBc was studied in the 65 dialysis patients and in 40 of the leukaemia patients. Three (4.6%) of the 65 dialysis patients and 12 (24.5%) of the 49 AML patients were anti-HCV positive whereas all of the ALL patients were seronegative. The total number of blood units transfused to 134 patients (data on two dialysis patients were not available) was 18,148, out of which 17,575 units had been transfused prior to the initiation of anti-HCV screening of blood donors. On the basis of the anti-HCV prevalence among blood donors and the incidence of post-transfusion hepatitis, the predicted number of seropositive patients was 11 and 18, respectively. Five of the 65 dialysis patients were anti-HBc positive, compared with only one of the 40 leukaemia patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228924 TI - Minimal role of hepatitis C virus infection in childhood liver diseases in an area hyperendemic for hepatitis B infection. AB - To investigate the role of hepatitis C virus (HCV) in childhood liver disease in Taiwan, an area hyperendemic for hepatitis B, we studied antibody to HCV (anti HCV) with a second generation enzyme immunoassay in 195 infants and children, including 96 hepatitis B surface antigen (HBsAg) positive children (66 with chronic hepatitis B, 23 children with hepatocellular carcinoma, and 7 with fulminant hepatitis B), 6 children with fulminant non-A, non-B hepatitis, 42 infants with neonatal hepatitis, 11 with biliary atresia, and 40 prospectively followed blood recipients. For comparison, another 748 apparently healthy children (from neonates to 12 years) were also screened for anti-HCV. The positive rate of anti-HCV was low in both apparently healthy children (0.13%) and patients with various liver disorders (0 to 4.4%) except fulminant hepatitis. The seropositive rate in 6 cases of non-A, non-B fulminant hepatitis was higher (16.7%) although the case number was too small. We conclude that HCV is generally not a major etiologic factor in the liver diseases of Taiwanese children. PMID- 8228925 TI - Persistence of Japanese encephalitis virus in the human nervous system. AB - Immunological and virological evidence for persistence of Japanese encephalitis virus (JEV) in the human nervous system is described in 16/323 (5%) laboratory confirmed cases of Japanese encephalitis. In 9/16 patients, JEV specific IgM antibodies were detected in the CSF even at 50-180 days after the onset of symptoms. Similarly, in 7/16 patients, apart from IgM antibodies, viral antigen was also present in the CSF beyond the third week of illness and in one patient it could be detected even at 117 days. Infectious virus could be isolated from the CSF beyond the third week of illness in 3/16 patients. In one patient, JEV was isolated from the CSF on three consecutive occasions at 90, 110, and 117 days after onset of clinical symptoms. These findings suggest that JEV persists in the nervous system of a small proportion of patients. PMID- 8228926 TI - Prevalence of hepatitis B, C, and D virus markers in Yemeni patients with chronic liver disease. AB - A serological survey for hepatitis B, C, and D markers was carried out in the Yemen Republic. Serum samples from 243 pregnant females, 294 male blood donors, and 108 patients with chronic liver disease were examined. Hepatitis B surface antigen (HBsAg) was found in 18.5% healthy individuals and 24.1% patients with chronic liver disease (P = 0.03). Evidence of any marker for hepatitis B virus (HBV) infection was found in 59.8% healthy individuals and 75.9% of patients with chronic liver disease (P = 0.0016). HBeAg was detected in 32.1% of the HBsAg positive pregnant females, indicating that vertical transmission probably plays a part in forming the pool of HBV carriers. Vaccination against HBV as part of the extended programme of immunisation (EPI) is recommended. Antibodies to hepatitis D were found in only 2 of 100 HBsAg-positive sera. Antibodies to hepatitis C (anti-HCV) were found in 2.1% healthy individuals and 21.5% patients with chronic liver disease (P = 0.0001). These results indicate that hepatitis B is hyperendemic in the Yemen Republic but that hepatitis D is very uncommon. The prevalence of anti-HCV is higher than in Europe and similar to neighbouring Arab countries. Infection with both HBV and HCV are important causes of chronic liver disease in the Yemen Republic. PMID- 8228927 TI - Detection of IgA class antibody to hepatitis E virus in serum samples from patients with hepatitis E virus infection. AB - A newly developed assay for IgA class antibody to hepatitis E virus (IgA anti HEV) was used to study 145 serum samples collected during an outbreak of an enterically transmitted hepatitis that occurred in 3 villages in the lower Shebeli region of Southern Somalia between January, 1988 and November, 1989. A total of 52.4% of the afflicted patients were found positive for IgA anti-HEV, and 73.1% of these were also positive for IgM. Both antibodies disappeared during the convalescence period. Similar results were also seen in serum obtained from sporadic cases of acute waterborne hepatitis in Pakistan. PMID- 8228928 TI - Comparative analysis of the restriction endonuclease profiles of the Dumas and Singapore strains of varicella-zoster virus. AB - The incidence of varicella in Singapore has been increasing since 1984. In 1991, 17,930 cases were reported in a population of about 3 million. A serological survey completed in 1990 demonstrated that only 43% of the cohort had antibodies to varicella-zoster virus (VZV), indicating inadequate herd immunity. To exclude novel VZV strains, representative VZV isolates from 9 chicken pox and 4 zoster patients were characterised by restriction endonuclease analysis. DNAs were extracted from viral isolates propagated in MRC5 human embryo lung cells and were digested separately with BglII, EcoRI, PstI, SalI, and XbaI enzymes. The cleavage profiles of these VZV strains derived from both chicken pox and zoster lesions revealed no distinct differences. This observation implies that the current upsurge of chicken pox most likely stems from closely related VZV genotypes infecting a susceptible population with insufficient herd immunity. Comparison of the restriction fragments of the Singapore and the Dumas strains revealed polymorphisms of the SalI-D, SalI-E, and XbaI-I fragment lengths, which correlated with variable regions I, II, and III of the VZV genome, thereby representing geographically distinct genotypic variants of VZV. PMID- 8228929 TI - Frequent isolation of human herpesvirus 7 from saliva samples. AB - Human herpesvirus 7 (HHV-7) was isolated frequently from saliva specimens. The isolation rates were 81% (13/16) in adults, 70% (7/10) in children over 1 year old, and none (0/7) in children less than 1 year old, respectively, indicating that infection of HHV-7 occurs during early infancy and the virus shedding rate after infection is very high. Human herpesvirus 6 (HHV-6) was not isolated from saliva specimens although some studies on isolation of HHV-6 from saliva was reported previously. PMID- 8228930 TI - The E2 region of HPV 16 in relation to different types of cervical lesions. AB - The presence of the E2 region of HPV 16 was determined by the polymerase chain reaction in biopsy samples with different histopathological diagnoses. The absence of the E2 gene was observed in 17 of 30 cases examined. The E2 deletions were not confined exclusively to invasive carcinomas or to integrated viral forms. In the majority of cases, the course of cervical lesions was consistent with the state of the E2 gene. PMID- 8228931 TI - Genomic typing of BK virus in clinical specimens by direct sequencing of polymerase chain reaction products. AB - Two hundred and twelve urine specimens, from several clinical groups, were examined for BK virus (BKV) using the polymerase chain reaction (PCR) to detect the VP1 region of BKV DNA. Positive results were obtained on 14 specimens from 44 post-transplant patients (31.8%), 10 specimens from 39 pregnant women (25.6%), and 5 specimens from 100 children (5%) but not on any specimens from 29 laboratory staff. Twenty-eight of the amplified BKV genomes, 19 from urine specimens, eight from culture fluid of inoculated tissue, and also one from a throat washing were directly sequenced from single-stranded templates immobilized via a biotinylated primer; it was possible to assign all to one of the four subtypes of BKV which had previously been identified on the basis of variation in nucleotide sequence of the VP1 region. Serological subgroup classification correlated with the genomic subtyping results in 21 of the isolates. The distribution of the BKV subtypes and the clinical status of the infected individuals are discussed. PMID- 8228932 TI - Two successive outbreaks of SRSV-associated gastroenteritis in South Africa. AB - Two successive outbreaks of gastroenteritis in South Africa were investigated to identify the aetiological agents. Some patients were involved in both outbreaks. Enteropathogenic bacteria or parasites were not evident in either outbreak. Small round structured viruses (SRSVs) were demonstrated in both outbreaks by direct electron microscopy. SRSV UK3/Hawaii virus was identified by immune electron microscopy as the causative agent in the first outbreak. Using new recombinant Norwalk virus (rNV) immunoassays for antibodies and antigen, Norwalk virus was implicated in the second outbreak. Preexisting antibodies to Norwalk virus were not protective and there was no cross protection between Hawaii and Norwalk viruses. There was no anamnestic response to Norwalk virus following the SRSV UK3/Hawaii outbreak although those affected had preexisting antibodies to Norwalk virus. To our knowledge, this is the first definitive diagnosis of SRSV associated gastroenteritis in South Africa. PMID- 8228933 TI - Cytomegalovirus recurrence in seropositive pregnant women attending obstetric clinics. AB - To understand reasons for cytomegalovirus (CMV) recurrence, a cohort of 350 CMV seropositive pregnant women attending obstetric clinics in Taiwan was examined for cervical or urinary CMV shedding. Urine specimens were collected from 350 women and cervical secretion specimens were collected from 220 women. We measured the association of various factors with CMV recurrence, which was defined as viral shedding identified by the presence of a CMV-specific gene sequence amplified by the polymerase chain reaction in seropositive individuals. CMV recurrence status was independently associated with a sexual activity composite variable, which was defined by three sexual activity indicators: age at first sexual intercourse, total years of sexual experience, and average frequency of sexual intercourse prior to pregnancy. Pregnant women with a history of genital tract infection were more likely than women without such history to experience cervical CMV recurrence. Similarly, pregnant women with previous urinary tract infections were more likely to experience urinary CMV recurrence. The findings indicate that multiple exposure to CMV by sexual activity prior to pregnancy is an important determinant of CMV recurrence during pregnancy. PMID- 8228934 TI - Immunogenicity of 20 micrograms of recombinant DNA hepatitis B vaccine in healthy neonates: a comparison of three different vaccination schemes. AB - The immunogenicity of a full dose (20 micrograms) of recombinant DNA yeast derived hepatitis B vaccine (Engerix-B) was assessed in healthy neonates in order to compare three candidate vaccination schemes. After randomization 162 newborns of hepatitis B surface antigen (HBsAg) negative mothers entered the study. Neonates received hepatitis B vaccine according to a four-dose vaccination scheme starting either at month 3 (scheme I: months 3, 4, 5, and 11) or at birth (scheme III: months 0, 1, 2, and 11). Another group of neonates received hepatitis B vaccine according to a three-dose scheme starting at birth (scheme II: months 0, 1, and 6). Serious adverse reactions were not observed; 2.5% of the vaccinated newborns suffered mild transient local symptoms. The vaccine was highly immunogenic irrespective of vaccination scheme; all infants developed anti-HBs levels > or = 10 IU/L, 97% > or = 100 IU/L. The immunogenicity of hepatitis B vaccine after primary and booster vaccinations, administered in the four-dose scheme started at birth, was significantly higher (P < 0.05) than in the three dose scheme started at birth. Hepatitis B vaccination according to the four-dose scheme started at month 3 produced significantly higher (P < 0.05) antibody levels in comparison to the four-dose scheme started directly after birth. This study showed that a four-dose hepatitis B vaccination scheme starting at month 3 resulted in the highest antibody levels of the three schemes investigated and can be recommended for incorporation in the Expanded Programme on Immunization in The Netherlands. PMID- 8228935 TI - Is there a relationship between malnutrition, inflammation, and post-vaccinal antibody response to influenza viruses in the elderly? AB - This study was designed to explore the relationship between malnutrition, inflammation, and the specific antibody response after influenza vaccination in the elderly. Eighty-two aged subjects, immunized annually against influenza with a trivalent inactivated vaccine, were evaluated for 9 protein markers (albumin, thyroxin-binding prealbumin, transferrin, immunoglobulins (Ig) G, M, and A, orosomucoid, haptoglobin, and C reactive protein) and for their antibody response to influenza viruses in comparison to 29 younger adults who received the same vaccine and 21 unvaccinated adults. IgM and nutritional markers were significantly reduced in the aged as compared to controls, while the opposite pattern was seen for IgA and inflammatory markers. No difference was observed between the elderly and the controls with regard to the antibody response to influenza virus after vaccination. Reciprocally, influenza immunization had no influence on the levels of the protein variables. These results suggest that the protein status does not play an important role in the antibody response to influenza vaccination in the elderly, a fact which could be related to the slight involvement of cellular immunity in the defense against influenza reinfection. PMID- 8228936 TI - Immunoglobulin M antibody response to measles virus following primary and secondary vaccination and natural virus infection. AB - The use of IgM antibody detection for the classification of the primary and secondary measles antibody response in persons following primary and secondary vaccination and natural measles virus infection was examined. Of 32 nonimmune children receiving primary measles vaccination, 31 (97%) developed IgM antibodies, consistent with a primary antibody response. Of 21 previously vaccinated children with low levels of preexisting IgG antibodies who responded to revaccination, none developed detectable IgM antibodies, whereas 33 of 35 (94%) with no detectable preexisting IgG antibodies developed an IgM response. Of a sample of 57 measles cases with a prior history of vaccination, 55 (96%) had detectable IgM antibodies. Of these, 30 (55%) were classified as having a primary antibody response and 25 (45%) a secondary antibody response based on differences in their ratios of IgM to IgG antibodies. Differences in the severity of clinical symptoms between these 2 groups were consistent with this classification scheme. These findings suggest that 1) an IgM response follows primary measles vaccination in the immunologically naive, 2) an IgM response is absent on revaccination of those previously immunized, and 3) an IgM response may follow clinical measles virus infection independent of prior immunization status. PMID- 8228937 TI - Human papillomavirus type 16 variants isolated from vulvar Bowenoid papulosis. AB - Tissues from two cases of Bowenoid papulosis of the vulva were found to contain human papillomavirus (HPV) 16 DNA by Southern blot hybridization. Analysis of the hybridization pattern revealed differences in a restriction fragment of one specimen as compared to the HPV 16 DNA prototype. To investigate if these differences could interfere with the expression of such oncogenic viral genomes, the corresponding DNA fragments were cloned and further analyzed. After amplification by PCR and DNA sequencing, a 213 base pairs duplication was mapped in the long control region (LCR) of this HPV 16 variant. One single PCR fragment was obtained from the other Bowenoid papulosis, which is identical in size with the same region in the HPV-16 prototype. The duplication in the HPV-16 LCR analyzed in this study maps upstream of a region containing several regulatory elements. PMID- 8228938 TI - Detection of hepatitis C virus RNA in peripheral blood mononuclear cells and in saliva. AB - The nested polymerase chain reaction (PCR) technique was applied to investigate hepatitis C virus (HCV) RNA in the peripheral blood mononuclear cells (PBMCs), saliva, and serum of patients with chronic type C hepatitis. The specificity of the amplified products was analyzed and confirmed by agarose gel electrophoresis, Southern blot hybridization, and restriction endonuclease pattern analysis. HCV RNA was detectable in the PBMCs of 24% (12/50) of the patients. The HCV RNA detected in PBMCs was not due to the contamination from plasma, since no viral sequences could be detected in the third washing of PBMCs. Of the 12 patients with HCV RNA in PBMCs, five were negative for HCV RNA sequences in the serum. Thus the presence of HCV RNA in PBMCs was not strictly correlated to the results for sera. Among 25 patients with HCV RNA in their saliva, 18 were negative for PBMCs. Among 25 patients without HCV RNA in their saliva, five had HCV RNA in PBMCs. In conclusion, PBMCs are an extrahepatic target for chronic HCV infection. However, we do not suggest that PBMCs act as a vehicle for carrying HCV to saliva, since the presence of HCV RNA in PBMCs and in saliva was not closely correlated. PMID- 8228939 TI - Detection of hepatitis C virus genome in human serum by multi-targeted polymerase chain reaction. AB - A multi-targeted "hemi-nested" PCR (M-PCR) assay in which the primer pairs derived from the 5' non-coding (5'NC) and the nonstructural protein 3 (NS3) regions of HCV genome were concurrently used for amplification in order to compare the sensitivity and specificity of polymerase chain reaction (PCR) with different primer pairs in detecting hepatitis C virus (HCV) genome. Sera from patients with virus-associated liver diseases were examined for the presence of HCV RNA by the M-PCR method following reverse transcription to cDNA. The amplified products derived from both the 5'NC and the NS3 regions were detected in 28 (70%) of the 40 HCV RNA-positive samples. However, 12 samples (30%) were devoid of the signal of NS3-derived product. Sensitivity tests using serial dilutions of HCV RNA revealed that the 5'NC-derived band was still detectable in the 10(5)-fold diluted sample by the M-PCR method, yet the NS3-derived band could hardly be detected in the 10(4)-fold diluted sample. Thus, as previously demonstrated by a single-targeted "nested" PCR assay, the present study using the M-PCR assay has clearly shown that the 5'NC-derived primers are more sensitive and specific than the NS3-derived primers in detecting HCV RNA. PMID- 8228940 TI - Inactivation of hepatitis A virus by heat treatment in aqueous solution. AB - Hepatitis A virus infections have been reported recently among hemophilic patients in Italy and Germany, leading to speculation that infectious hepatitis A virus (HAV) might have been present in some factor VIII concentrates. In both cases, the implicated factor concentrates had been treated by a solvent/detergent method, which inactivates enveloped viruses but which would not be expected to inactivate HAV, a nonenveloped picornavirus. To determine whether HAV would be inactivated during pasteurization of factor VIII concentrate, an alternative method employed for virus inactivation, we determined the extent to which the infectivity of cell culture-adapted HAV, suspended either in cell culture medium or in a proprietary stabilizing buffer, was reduced by heat treatment at 60 degrees C for 10 hr. The titer of infectious HAV declined rapidly at 60 degrees C, but the stabilizer considerably delayed HAV inactivation. In cell culture medium, HAV was inactivated by > 3.6 log10 within 30 min, but 3.6 log10 inactivation of HAV was reached only after 6 hr in the presence of the stabilizer. Residual infectious HAV was present after even 10 hr of heat treatment in the stabilizer, indicating that < 5.2 log10 infectious HAV particles are inactivated under these conditions. In the presence of the stabilizer, HAV was significantly more stable than poliovirus type 1, which has been used to validate virus inactivation by pasteurization. We conclude that pasteurized factor VIII concentrate should pose little if any risk for transmission of HAV if pooled plasma used for its manufacture contained low levels of the virus. PMID- 8228941 TI - Long-term carriage of hepatitis C virus with normal aminotransferase after interferon treatment in patients with chronic hepatitis C. AB - Studies were undertaken to investigate whether interferon therapy could induce hepatitis C virus (HCV) carriage with normal serum alanine aminotransferase (ALT) values using an assay that combined reverse transcription and polymerase chain reaction. The subjects studied were 53 patients with chronic active hepatitis C who received interferon (alpha, 33 cases; beta, 20 cases) therapy. All were seropositive for HCV RNA prior to therapy. In all 22 complete responders, whose ALT levels fell to normal during therapy and for at least 24 weeks after therapy, HCV RNA became persistently negative except in two cases. The two had sustained viremia on treatment and for 1.0-1.5 years of follow-up, although their biochemical tests were normal. In 15 patients with a transient response in whom the disease recurred when interferon was stopped, HCV RNA was undetectable in 80% of the cases at the end of therapy, but the virus reappeared with subsequent elevation of ALT in all patients. However, 3 patients in this group had normal enzyme levels with viremia for 2.1-2.8 years of follow-up after acute deterioration of illness. In 16 patients who did not respond to interferon, HCV RNA was persistently positive during and after therapy. These findings suggest that interferon therapy induces a long-term carrier state of HCV infection with normal ALT levels in some patients. PMID- 8228942 TI - Detection of human papillomavirus L1 protein in condylomata acuminata from adults with defects in cell-mediated immunity. AB - Immunohistochemical assays for human papillomavirus (HPV) L1 protein, using antiserum directed against the L1 major capsid protein of bovine papillomavirus (anti-BPV serum), were performed on 101 condylomata acuminata biopsy samples from 47 men (40 of whom had intact cell mediated immunity [CMI], and 7 with conditions known to cause CMI defects), and 54 women (48 with normal CMI, and 6 with CMI defects). L1 protein was detected in 28% of all biopsies, including 20.5% of samples from patients with normal CMI and 76.9% of patients with CMI defects (P = .00002). For both males and females, L1 protein was detected significantly more often in samples from patients with CMI defects than those with normal CMI. Immunohistochemical assays of HPV 11-infected human foreskin implants grown in athymic mice were performed to optimize the conditions of the assay. Three dilutions of anti-BPV serum or preimmune rabbit serum were used. A 1:500 dilution provided readily interpretable results, while preimmune serum at this dilution did not stain to any significant degree. However, at 1:10 and 1:100 dilution, both the anti-BPV serum and preimmune serum caused an unacceptable amount of nuclear staining, making results uninterpretable. Defects in CMI may allow active viral particle assembly at an increased rate, as judged by detection of L1 protein in condylomata acuminata from these patients. PMID- 8228943 TI - Administration of inactivated and detergent-treated influenza virus to mice before virus challenge reduces mortality. AB - Formalin-inactivated virus (FIV) and the detergent-treated virus (DTV) preparations were tested for their ability to enhance the resistance of mice to experimental influenza infection. FIV (100 micrograms) was administered intravenously to mice. After 24 hr, animals were challenged with 5 LD50 dose intranasally. FIV-treated and non-treated (control) mice had 10% and 100% mortality, respectively. Similar results were obtained with the DTV (40 micrograms) preparation. The pulmonary virus titer of FIV-treated mice was lower when compared with the control. Mechanisms other than acquired immunity may have conferred the early resistance to virus infection in mice. PMID- 8228944 TI - Preliminary evidence that a trpE-HEV fusion protein protects cynomolgus macaques against challenge with wild-type hepatitis E virus (HEV). AB - Immunization of two cynomolgus macaques (cynos) with trpE-C2 protein, a trpE-HEV fusion protein that represents the carboxyl two thirds of the putative capsid protein, prevented development of biochemical evidence of viral hepatitis in these primates after challenge by wild-type HEV from either a Burmese or Mexican stool isolate. Neither of the immunized animals showed any elevation of alanine aminotransferase activity after challenge with wild-type HEV in marked contrast with the unimmunized (control) cynos. In the case of the Burmese HEV challenged cyno, the protective effect was complete with the animal failing to demonstrate any evidence of HEV infection. The immunized cyno challenged with Burmese HEV did not exhibit any HEV RNA in its stools or HEV antigen in its liver. The immunized cyno (#8902) challenged with Mexican virus exhibited HEV RNA in its stools and HEV antigen in its liver; however, microscopic examination of liver biopsy specimens from this cyno failed to detect histopathologic evidence of viral hepatitis. All of the animals (naive and immunized) developed anti-HEV IgM and IgG responses after HEV challenge. Our preliminary studies indicate that the trpE C2 protein is a promising candidate HEV vaccine. PMID- 8228945 TI - Personality and depression in Parkinson's disease. AB - The Parkinson's disease (PD) patient has been characterized as having a distinctive personality with introverted features. These personality traits are said to predate motor symptoms and are theorized to serve as a subtle clue to latent PD. To examine this hypothesis, we compared remote and current personality features in 35 PD subjects and 35 controls. Subjects' spouses completed a personality inventory (PI) characterizing patients' premorbid and current status. The premorbid PI of PD subjects differed from that of controls in being more "quiet," "generous," "cautious," and "even-tempered," and less "flexible." The characterization of the PD subjects' current personality differed greatly from reported premorbid personality features, i.e., significant change in 13 of 24 PI items. Personality inventory responses regarding both the PD subjects' premorbid and current personality correlated to symptoms of depression and disease severity. Cognition, tobacco use, alcohol consumption, and rural versus urban residency did not correlate with PI responses. We conclude that PD patients are apt to be viewed as introverts premorbidly, and, with disease onset, more striking personality changes are recognized. These perceptions appear to be closely linked to depressed affect and correlate with motor impairment to a lesser extent. PMID- 8228946 TI - Positive associations among dichotic listening errors, complex partial epileptic like signs, and paranormal beliefs. AB - The numbers of complex partial epileptic-like signs were moderately (rs = .50) associated with strength of paranormal beliefs but not conservative (religious) beliefs in a population of young males and females (13 to 20 years). Dichotic listening errors were also significantly correlated with both complex partial epileptic-like signs and paranormal, but not conservative, beliefs; the females demonstrated the strongest intercorrelations (rs between .54 and .63). These results support the hypotheses that dichotic listening errors reflect a continuum of complex partial epileptic phenomenology in the normal population and this process encourages the acquisition of paranormal but not traditional beliefs. PMID- 8228947 TI - Positive associations among dichotic listening errors, complex partial epileptic like signs, and paranormal beliefs. PMID- 8228948 TI - Birth weight and CT scan findings in chronic schizophrenic patients. AB - The relations of birth weight to CT scan measures of lateral ventricular size, third ventricular size, frontal cortical sulcal prominence, and parieto-occipital cortical sulcal prominence were examined in 24 chronic schizophrenic patients. In contrast to the inverse correlation between birth weight and ventricular size found among offspring of schizophrenic parents in the Danish High-Risk Study, and the predictions of the subsequent hypothesis, no significant relations between birth weight and CT scan measures were observed in this study. PMID- 8228949 TI - Evaluation of the ratings comprising the associated symptom criterion of DSM-III R generalized anxiety disorder. AB - In light of ongoing considerations for DSM-IV, the 18 ratings comprising the associated symptom criterion of DSM-III-R generalized anxiety disorder were evaluated. The interrater reliability and endorsement rates of each symptom were calculated from interview-based ratings of generalized anxiety disorder patients at four sites (total N = 204). Interrater reliability analyses indicated high agreement for the majority of the 18 symptoms. Regarding the frequency of symptom endorsement, significant Spearman correlations (range = .69 to .94) revealed marked consistency in the rates of symptom endorsement across the four sites. A list of "satisfactory" symptoms was generated from those meeting certain reliability and endorsement criteria. Seven symptoms were identified: irritability, restlessness, muscle tension, difficulty concentrating, sleep difficulties, feeling keyed up, and easy fatigability. Interestingly, all of the above belong to either the motor tension or vigilance and scanning clusters of the DSM-III-R associated symptom ratings. In contrast, symptoms belonging to the autonomic hyperactivity cluster were endorsed infrequently at all sites. Although additional research is needed (e.g., examination of the symptoms' discriminant validity), these findings essentially support revising the associated symptom criterion of generalized anxiety disorder in accord with option E3 in the DSM-IV Options Book. PMID- 8228950 TI - Posttraumatic stress disorder symptoms and precombat sexual and physical abuse in Desert Storm veterans. AB - The purpose of this research was to study the association between precombat sexual and physical abuse and combat-related posttraumatic stress disorder (PTSD) symptoms in a clinical sample of male and female Desert Storm veterans. Two hundred ninety-seven veterans provided data on precombat sexual and physical abuse, precombat psychiatric problems, sociodemographics, Desert Storm combat exposure, and PTSD symptomatology using the Mississippi Scale. Men reported significantly higher levels of combat exposure, and women described significantly more frequent precombat abuse. Precombat-abused veterans reported more frequent precombat psychiatric histories. Analysis of covariance revealed that gender significantly modified the impact of precombat abuse on combat-related and other PTSD symptomatology after adjusting for precombat psychiatric history and level of combat exposure. Specifically, females describing precombat abuse reported much greater PTSD symptomatology than did females denying precombat abuse. These results in conjunction with previous research suggest that a relationship between precombat abuse and combat-related PTSD may exist. Prospective, longitudinal studies of both men and women are needed. PMID- 8228951 TI - Assessing alcoholic denial. Further examination of the Denial Rating Scale. AB - The reliability, validity, and heuristic value of Goldsmith and Green's (J. Nerv. Ment. Dis. 176:614-620, 1988) Denial Rating Scale (DRS) are examined in an inpatient treatment setting. This replication study includes 647 cases. The same strong relationship between clinical change and program completion as found by Goldsmith and Green is identified. An ordinal predictive validity for the DRS regarding program completion that was not found by Goldsmith and Green is also identified. Utility for improving patient treatment plans through DRS results is identified. The heuristic value of the DRS in helping an alcoholism counseling staff maintain treatment focus is discussed. It is recommended that treatment centers incorporate systematic assessment of denial over the course of treatment as a means to facilitate staff focus on the primacy of addiction. PMID- 8228952 TI - Insight, overvalued ideation, and delusional thinking in body dysmorphic disorder: theoretical and treatment implications. PMID- 8228953 TI - A comparison of weight changes with fluoxetine, desipramine, and amitriptyline: a retrospective study of psychiatric inpatients. PMID- 8228954 TI - Bulimia nervosa in outpatients with migraine: a pilot study. PMID- 8228955 TI - Naltrexone abuse and risk of suicide. PMID- 8228956 TI - Nocturnal limb movements in conversion paralysis. PMID- 8228957 TI - Discrimination between anxiety and depression. PMID- 8228958 TI - Putnam, Jackson, and posttraumatic stress disorder. PMID- 8228959 TI - Planar and vertical induction of anteroposterior pattern during the development of the amphibian central nervous system. AB - In amphibians and other vertebrates, neural development is induced in the ectoderm by signals coming from the dorsal mesoderm during gastrulation. Classical embryological results indicated that these signals follow a "vertical" path, from the involuted dorsal mesoderm to the overlying ectoderm. Recent work with the frog Xenopus laevis, however, has revealed the existence of "planar" neural-inducing signals, which pass within the continuous sheet or plane of tissue formed by the dorsal mesoderm and presumptive neurectoderm. Much of this work has made use of Keller explants, in which dorsal mesoderm and ectoderm are cultured in a planar configuration with contact along only a single edge, and vertical contact is prevented. Planar signals can induce the full anteroposterior (A-P) extent of neural pattern, as evidenced in Keller explants by the expression of genes that mark specific positions along the A-P axis. In this review, classical and modern molecular work on vertical and planar induction will be discussed. This will be followed by a discussion of various models for vertical induction and planar induction. It has been proposed that the A-P pattern in the nervous system is derived from a parallel pattern of inducers in the dorsal mesoderm which is "imprinted" vertically onto the overlying ectoderm. Since it is now known that planar signals can also induce A-P neural pattern, this kind of model must be reassessed. The study of planar induction of A-P pattern in Xenopus embryos provides a simple, manipulable, two-dimensional system in which to investigate pattern formation. PMID- 8228960 TI - Induction and axial patterning of the neural plate: planar and vertical signals. AB - In this review I summarize recent findings on the contributions of different cell groups to the formation of the basic plan of the nervous system of vertebrate embryos. Midline cells of the mesoderm--the organizer, notochord, and prechordal plate--and midline cells of the neural ectoderm--the notoplate and floor plate- appear to have a fundamental role in the induction and patterning of the neural plate. Vertical signals acting across tissue layers and planar signals acting through the neural epithelium have distinct roles and cooperate in induction and pattern formation. Whereas the prechordal plate and notochord have distinct vertical signaling properties, the initial anteroposterior (A-P) pattern of the neural plate may be induced by planar signals originating from the organizer region. Planar signals from the notoplate may also contribute to the mediolateral (M-L) patterning of the neural plate. These and other findings suggest a general view of neural induction and axial patterning. PMID- 8228961 TI - Mechanisms of early neurogenesis in Drosophila melanogaster. AB - The neuroectoderm of insects contains an initially indifferent population of cells which during later development will give rise to the progenitor cells of the neural and epidermal lineages. Experimental evidence indicates that cellular interactions determine which cells will adopt each one of these fates. Transplantation experiments suggest that a signal with neuralising character is required to stabilize the primary neural fate in 25% of all the neuroectodermal cells, which will develop as neuroblasts, and that an epidermalising signal contributes to suppress the neural fate in the remaining 75% of the cells, allowing in this way their development as epidermal progenitor cells. The invoked cell interactions are assumed to be mediated by the products of several genes forming a complex, not yet well understood network of interrelationships. Elements of this network are the proteins encoded by Delta and Notch, which appear to convey the regulatory signals between the cells; the proteins encoded by the achaete-scute gene complex, which regulate neural development; and the proteins encoded by the Enhancer of split gene complex, which give neuroectodermal cells access to epidermal development. PMID- 8228962 TI - Fate and potentialities of the avian mesencephalic/metencephalic neuroepithelium. PMID- 8228963 TI - Pax: gene regulators in the developing nervous system. AB - In recent years, the discovery of Pax genes in mouse has played an invaluable role in furthering our understanding in mouse developmental processes and disorders. To date, eight murine paired box-containing genes have been cloned. Seven of these exhibit a distinct spatiotemporal expression pattern in the developing nervous system implying a role in the regional specification of the developing spinal cord and brain. The Pax genes encode for sequence-specific DNA binding transcription factors that play a key role in embryonic development. Three of these developmental control genes are altered in mutant mice and two are associated with human diseases. Disruption of these Pax genes leads to abnormalities in neural crest derivatives, neuroectoderm, sclerotome or myotome derived tissues. Disruption of the Pax-3 gene causes the Splotch phenotype in mice and Waardenburg syndrome in humans. Pax-6 mutations result in Small eye mice and the human genetic disorder aniridia. The Pax-1 gene is mutated in undulated mice. Pax proteins can transform cells in culture which then form tumours following injection in nude mice. Consistent with this activity, PAX3 has been recently implicated in the generation of the tumour alveolar rhabdomyosarcoma. PMID- 8228964 TI - Position, guidance, and mapping in the developing visual system. AB - Positional identity in the visual system affects the topographic projection of the retina onto its central targets. In this review we discuss gradients and positional information in the retina, when and how they arise, and their functional significance in development. When the axons of retinal ganglion cells leave the eye, they navigate through territory in the central nervous system that is rich in positional information. We review studies that explore the navigational cues that the growth cones of retinal axons use to orient towards their target and organize themselves as they make this journey. Finally, these axons arrive at their central targets and make a precise topographic map of visual space that is crucial for adaptive visual behavior. In the last section of this review, we examine the topographic cues in the tectum, what they are, when, and how they arise, and how retinal axons respond to them. We also touch on the role of neural activity in the refinement of this topography. PMID- 8228965 TI - Touch receptor development and function in Caenorhabditis elegans. AB - Mutations causing a touch-insensitive phenotype in the nematode Caenorhabditis elegans have been the basis of studies on the specification of neuronal cell fate, inherited neurodegeneration, and the molecular nature of mechanosensory transduction. PMID- 8228966 TI - The history of the discovery of the nerve growth factor. AB - The Nerve Growth Factor (NGF) is the progenitor of a family of growth factors which is still expanding. The history of its discovery is very colorful; it is a rare combination of scientific reasoning, intuition, fortuities, and good luck. In addition, I believe that the collaboration of three scientists with very different backgrounds contributed to the success: I had grown up in a laboratory of experimental embryology, Dr. Levi-Montalcini came from neurology, and Dr. Stanley Cohen was from biochemistry. The decision where to begin the history of a discovery is always arbitrary. I shall give my reasons why I begin this story with my wing bud extirpations on chick embryos and the analysis of the effects of the operation on the development of spinal nerve centers, published in 1934. Of course, I am aware of the fact that the analysis of neurogenesis had been pioneered by Dr. R. G. Harrison and his students at Yale University since the beginning of this century. It should be mentioned that their experiments had been done on amphibian embryos. My own interest in problems of neurogenesis dates back to my Ph.D. thesis in the Zoology Department of Professor H. Spemann at the University of Freiburg in (the Federal Republic of) Germany; it dealt with the influence of the nervous system on the development of limbs in frog embryos. After I had obtained some inconclusive results I did the crucial experiment of producing nerveless legs. I removed the lumbar part of the spinal cord and the spinal ganglia before the outgrowth of nerve fibers. The nerveless legs developed normally in every respect, but the muscles atrophied eventually. PMID- 8228967 TI - Regenerating retinal fibers display error-free homing along undamaged normal fibers. AB - After crushing one optic nerve in a bony fish, retinal fibers regenerate to both tecta. Anterograde labelling indicates that the ipsilaterally regenerating fibers have a rather straight growth, apparently along the undamaged fibers of the contralateral retina. In contrast, the contralaterally regenerating fibers deviate widely from a straight course. Retrograde labelling shows a mirror symmetric distribution of regenerated ipsilateral and resident contralateral ganglion cells in a comparable annulus. In contrast, ganglion cells in the regenerated contralateral retina show no topological order after comparable small Dil applications to the ventrolateral tectum. These data suggest that regenerating fibers can orient on the undisturbed, contralateral fibers. PMID- 8228968 TI - Temporal patterning of song production: participation of nucleus uvaeformis of the thalamus. AB - Birdsong is a learned vocal behavior used in intraspecific communication. The motor pathway serving learned vocalizations includes the forebrain nuclei NIf, HVC, and RA; RA projects to midbrain and brain stem areas that control the temporal and acoustic features of song. Nucleus Uvaeformis of the thalamus (Uva) sends input to two of these forebrain nuclei (NIf and HVC) but has not been thought to be important for song production. We used three experimental approaches to reexamine Uva's function in adult male zebra finches. (1) Electrical stimulation applied to Uva activated HVC and the vocal motor pathway, including tracheosyringeal motor neurons that innervate the bird's vocal organ. (2) Bilateral lesions of Uva including the dorso-medial portion of the nucleus affected the normal temporal organization of song. (3) Chronic multiunit recordings from Uva during normal song and calls show bursts of premotor activity that lead the onset of some song components, and also larger bursts that mark the end of complete song motifs. These results implicate Uva in the production of learned vocalizations, and further suggest that Uva contributes more to the temporal structure than to the acoustic characteristics of song. PMID- 8228969 TI - Natural fluctuation and gonadal hormone regulation of astrocyte immunoreactivity in dentate gyrus. AB - The number and the surface density of cells immunoreactive for the specific astrocytic marker glial fibrillary acidic protein (GFAP), were evaluated in both the hilus of the dentate gyrus and the granular layer of the vermis of the cerebellar cortex of adult female rats during the different phases of the estrous cycle, after ovariectomy and after the pharmacological administration of estradiol and/or progesterone to ovariectomized rats. Although no significant differences were detected in the number of immunoreactive cells among the different experimental groups studied, their surface density showed significant changes in the hilus of the dentate gyrus. The surface density of immunoreactive cells was increased in the afternoon of proestrus and on the morning of estrus compared to the morning of proestrus, diestrus, and metestrus, was decreased after ovariectomy, and showed a dose-dependent increase in ovariectomized rats injected with 17 beta estradiol (1, 10, or 300 micrograms/rat), alone or in combination with progesterone (500 micrograms/rat). In contrast, it was not affected by the administration of 17 alpha estradiol (300 micrograms/rat). The surface density of immunoreactive cells was significantly increased over control values by 5 h after the injection of 17 beta estradiol (300 micrograms/rat) and as early as 1 h after the administration of progesterone. The separate injection of either 17 beta estradiol or progesterone had smaller effects on the surface density of immunoreactive cells than did the administration of both hormones together. The surface density of GFAP-immunoreactive cells reached maximal values by 24 h after the administration of 17 beta estradiol and/or progesterone and returned to control levels by 48 h after the combined injection of progesterone and 17 beta estradiol, while in the rats that were injected with only one of the two hormones, the surface density of immunoreactive cells remained over control values for at least 9 days. No such hormonal effects on GFAP-immunoreactive cells were observed in the cerebellar cortex. PMID- 8228970 TI - Intracellular partitioning of androgen receptor immunoreactivity in the brain of the male Syrian hamster: effects of castration and steroid replacement. AB - The effect of castration and steroid replacement on the intracellular partitioning of the androgen receptor in the brain of the male Syrian hamster was determined using immunocytochemistry. Androgen receptors were visualized using the PG-21 antibody (G. S. Prins) on 40-microns coronal brain sections from hamsters perfused with 4% paraformaldehyde with or without 0.4% glutaraldehyde. Control studies confirmed antibody specificity in gonad-intact and castrate males. In the normal adult male, androgen receptor immunocytochemistry reveals intense staining confined to the cell nucleus. Castration caused a gradual increase in cytoplasmic labelling within 2 weeks, accompanied by a reduction in nuclear staining intensity in androgen receptor-containing neurons throughout the brain. Cytoplasmic androgen receptor staining was eliminated after treatment of orchidectomized males for only 8 h with exogenous testosterone. Likewise, long term exposure to testosterone and dihydrotestosterone, a nonaromatizable androgen, maintained nuclear androgen receptor immunoreactivity. However, exposure to low physiologic concentrations of estrogen was not effective in this regard. In addition, we determined that nuclear androgen receptor immunoreactivity decreases in response to inhibitory short-day photoperiod, but without an increase in cytoplasmic immunostaining. This appears to be due to the decrease in androgen production by the testis, rather than a direct photoperiodic effect, because testosterone supplementation to short-day males restored the intensity of nuclear androgen receptor immunoreactivity to levels comparable to those in the intact male. These findings are compatible with a new model for the intracellular localization of androgen receptors, in which a subset of unoccupied receptors is located in the cell cytoplasm in the absence of ligand. They further demonstrate the repartitioning of such cytoplasmic receptors, thereby confirming and extending previous observations using biochemical techniques on the regulation of neuronal androgen receptors. PMID- 8228971 TI - Immunocytochemical localization of sodium channels in an insect central nervous system using a site-directed antibody. AB - Antibodies to channel proteins and specific peptide sequences have been previously used to localize voltage-activated sodium channels in the rat brain. Here we describe the first localization of sodium channels in an insect nervous system using a site-directed antibody. The mesothoracic ganglion of the cockroach was stained with an antibody to the highly conserved SP19 sequence. Antibody labelling was visualized by light microscopy using the avidin/biotin method on wax sections, and transmission electron microscopy of immunogold-labelled thin sections. Central ganglia of insects contain clearly separated regions of cell bodies, synaptic neuropil, axon tracts, and nerves. Antibody staining by light microscopy was limited to neurons, and was intense in axons throughout the ganglion and nerves. Staining was also strong in the cytoplasm, but not the nuclei, of many neuronal cell bodies. Neuropil regions were relatively lightly labelled. These findings can be correlated with the known electrophysiology of the ganglion. Electron microscopy detected sodium channels in areas surrounding axons, probably including axon membranes and enveloping glial cell membranes. Axonal mitochondria were also heavily labelled, suggesting a sodium channel transport function for these organelles. PMID- 8228972 TI - Developmental study of the expression of B50/GAP-43 in rat retina. AB - B50/GAP-43 has been implicated in neural plasticity, development, and regeneration. Several studies of axonally transported proteins in the optic nerve have shown that this protein is synthesized by developing and regenerating retinal ganglion cells in mammals, amphibians, and fish. However, previous studies using immunohistochemistry to localize B50/GAP-43 in retina have shown that this protein is found in the inner plexiform layer in adults. Since the inner plexiform layer contains the processes of amacrine cells, ganglion cells, and bipolar cells to determine which cells in the retina express B50/GAP-43, we have now used in situ hybridization to localize the mRNA that codes for this protein in the developing rat retina. We have found that B50/GAP-43 is expressed primarily by cells in the retinal ganglion cell layer as early as embryonic day 15, and until 3 weeks postnatal. Some cells in the inner nuclear layer, possibly a subclass of amacrine cells, also express B50/GAP-43 protein and mRNA; however, the other retinal neurons-bipolar cells, photoreceptors, and horizontal cells express little, if any, B50/GAP-43 at any stage in their development. Early in development, the protein appears in the somata and axons of ganglion cells, while later in development, B50/GAP-43 becomes concentrated in the inner plexiform layer, where it continues to be expressed in adult animals. These results are discussed in terms of previous proposals as to the functions of this molecule. PMID- 8228973 TI - Injury-associated induction of GAP-43 expression displays axon branch specificity in rat dorsal root ganglion neurons. AB - Peripheral nerve injury results in the increased synthesis and axonal transport of the growth-associated protein GAP-43 in dorsal root ganglion (DRG) neurons, coincident with regenerative growth of the injured peripheral axon branches. To determine whether the injury-associated signalling mechanism which leads to GAP 43 induction also operates through the central branches of DRG axons, we used immunocytochemistry to compare the expression of GAP-43 in adult rat DRG neurons 2 weeks after dorsal root crush lesions (central axotomy) or peripheral nerve crush lesions (peripheral axotomy). In uninjured ganglia, a subpopulation of smaller DRG neurons expresses moderate levels of GAP-43, whereas larger neurons generally do not. At 2 weeks following peripheral axotomy, virtually all axotomized neurons, large and small, express high levels of GAP-43. At 2 weeks following dorsal root lesions, no increase in GAP-43 expression is detected. Thus, the injury-associated up-regulation of GAP-43 expression in DRG neurons is triggered by a mechanism that is responsive to injury of only the peripheral, and not the central, axon branches. These findings support the hypothesis that GAP-43 induction in DRG neurons is caused by disconnection from peripheral target tissue, not by axon injury per se. PMID- 8228974 TI - Dendritic reduction in Passover, a Drosophila mutant with a defective giant fiber neuronal pathway. AB - The jump response to a light-off startle stimulus in Drosophila melanogaster occurs when the Giant Fiber (GF), a neuron descending from the brain to the thorax, drives the jump (tergotrochanteral) muscle motorneuron (TTMn). Nonjumping mutants have been isolated in which this response is disrupted. Flies bearing the X-chromosome mutation Passover (Pas) fail to jump in response to a light-off stimulus, and electrical stimulation of the GF in the brain no longer elicits the normal response in the TTM. We have used retrograde HRP labelling to examine the TTMn motorneuron in wild-type flies and in a variety of newly identified Pas alleles. In wild type the medial branch (MB) of the TTMn has an extensive region of apposition with the GF. In Pas alleles, there is a general reduction in anterior-posterior (A-P) extent of the medial branch but not of the posterior branch. Nevertheless, Pas alleles usually leave the TTMn close enough to the GF so that contact would not be precluded. In flies carrying a particular deficiency of Pas, Df(1) 16-3-22, including Pas/Df(1) 16-3-22 heterozygotes, there can be extensive growth of the medial-branch including a contralateral projection; these heterozygotes have more than the normal amount of overlap between the GF and the TTMn. This phenotype, originally ascribed to Pas mutants, is associated with Df(1) 16-3-22, but not with other deletions of the Pas gene. The driving of the TTMn by the GF is defective in mutant genotypes with extensive medial branches as well as in mutants where GF-TTMn contact is reduced. The fact that the TTMn grows into its normal synaptic region in mutant genotypes, but the GF pathway functions abnormally suggests that pathfinding by the TTMn is not impaired. It is more likely that the Pas mutation disrupts cell recognition, synaptogenesis, or synaptic function in the TTMn or its presynaptic partners. PMID- 8228975 TI - Synaptic repression at crayfish neuromuscular junctions. I. Generation after partial target area removal. AB - Synaptic repression, the inability of synaptic junctions to generate normal-sized postsynaptic potentials under normal physiological conditions, is reported here for crayfish neuromuscular synapses. The synapses in the superficial flexor muscle system of the crayfish change their efficiency in generating a postsynaptic response as a result of a specific alteration in their immediate environment. When the superficial flexor nerve is cut halfway into the target muscle field and the lateral muscle fibers are removed, the intact medial synapses do not generate normal-sized junction potentials (JP) at the 17 degrees 19 degrees C temperature of the Ringers solution. JPs cannot be recorded in 83% of the muscle fibers at 2 weeks after the operation and of the few JPs that can be detected, 80% are smaller than 1 mV in size. By 8 weeks after the operation, JPs were detected in 55% of the muscle fibers, and now only 46% of these are smaller than 1 mV. When the lateral muscle fibers are left in place during the original operation, providing a target area for the cut nerve to grow into, JPs were then detected in 60%-80% of all medial fibers at all time periods after the operation; their size profile, with 10%-25% of the muscle fibers having JP's less than 1 mV, was similar to control values. These results suggest that the efficiency of these synaptic contacts become affected as a result of partial axotomy and removal of the target area of the cut branches of the axons. PMID- 8228976 TI - Kinetic and thermodynamic studies of the copper (II) and nickel(II) complexes of glycylglycyl-L-histidine. AB - The protonation constants of the tripeptide glycylglycyl-L-histidine (L-) have been determined at 25 degrees C and I = 0.1 mol dm-3 as log K 8.06, 6.82, and 2.80. Complexation with copper(II) can be represented by the series of equilibria [formula: see text] in the case of nickel(II) only the species [NiLH]2+, [NiL]+, and [NiLH-2]- are of importance with log beta 111 = 11.33(2); log beta 110 = 4.74(6), and log beta 11-2 = -6.93(1). The tripeptide acts as a quadridentate ligand to give complexes with copper and nickel with an amino group, two deprotonated amide groups and an imidazole pyridine nitrogen (Im-N3) as donors. At 1:1 ligand-to-metal ratios the purple copper(II) complex [CuLH-2]- is essentially 100% abundant above pH 7 and the planar yellow [NiLH-2]- above pH 8. The displacement of the tripeptide ligand from the nickel(II) complex by L histidine has been studied kinetically over the pH range 7-8. There is a small solvolytic reaction and a reaction which is first-order in the hydrogen ion concentration. Under the experimental conditions employed, the reaction is essentially independent of the L-His concentration and displacement occurs by a proton-assisted nucleophilic pathway with rate-determining cleavage of the first nickel(II)-N(peptide) bond. PMID- 8228977 TI - Palladium (II) compounds of putrescine and spermine. Synthesis, characterization, and DNA-binding and antitumor properties. AB - The reaction of putrescine (Put) with K2PdCl4 and PdCl2 resulted in the synthesis of compounds of formula [PutH2][PdCl4] and [Pd2Cl4(Put)2]. Compounds of formula [PdCl2(SpermH2)][PdCl4] and [Pd2Cl4(Sperm)] have been also synthesized by reaction of spermine (Sperm) with K2PdCl4. The structure of all these compounds has been analyzed by IR and 1H NMR. UV and CD spectroscopic data have shown that all the Pd(II)-polyamine compounds synthesized induce conformational changes in the circular forms of plasmid DNA. Determinations by electrophoresis in agarose gels of the mobility of the DNA in drug:DNA complexes indicated that only the Pd(II)-putrescine compounds have the ability to induce significant conformational changes in the covalently closed circular (ccc) form of the pUC8 plasmid DNA. The Pd(II)-putrescine and Pd(II)-spermine compounds were also assayed for in vitro antiproliferative activity against MDA-MB 468 and HL-60 human cancer cells. The results suggest that the putrescine complexes may be regarded as potential antitumor agents because the ID50 value of all of the Pd(II)-putrescine complexes is twofold lower than the ID50 of cis-DDP. Our data also show that, on the other hand, the Pd(II)-spermine compounds have low antiproliferative activity. PMID- 8228978 TI - X-ray crystal structure of tetrachloro(trans-d,l-1,2-diaminocyclohexane) platinum(IV) complex: a potential anticancer agent. AB - Tetrachloro(trans-d,l-1,2-diaminocyclohexane)platinum(IV) complex (tetraplatin or ormaplatin) has been recognized as a potential anticancer agent. We have determined the crystal structure of this compound by x-ray single crystal diffraction. It has a unique space group and rare molecular packing. Parameters are as follows: space group R3c (rhombohedral), a = 26.425(4) A, alpha = 54.50(1) degrees; V = 11375A3, Z = 36. The coordination about Pt atoms is a slightly distorted octahedron owing to the presence of the geometrically strained five membered ring (avg. N-Pt-N angle = 82.9 degrees). An intricate network of intermolecular hydrogen bonds holds the crystal lattice together. Three independent ormaplatin molecules surround each water molecule, forming the three shortest A...B contacts and thus the three strongest hydrogen bonds. PMID- 8228979 TI - Purification, composition, charge, and molecular weight of the FeMo cofactor from Azotobacter vinelandii nitrogenase. AB - A procedure has been developed for purifying NMF and NMF/DMF solutions of the FeMo cofactor (FeMoco) derived from the molybdenum iron protein of nitrogenase. This procedure consists of anaerobic chromatography of FeMoco solutions on two consecutive anaerobic molecular sizing columns followed by electrophoretic migration through a third sizing column. FeMoco prepared by this procedure is homogeneous as evidenced by chromatographic, electrophoretic, and compositional criteria. The minimal elemental composition was found to be MoFe6S6 using chemical colorimetric, inductively coupled plasma (ICP), and proton induced x-ray emission (PIXE) analytical procedures. Molecular weight measurements of NMF and DMF solutions of FeMoco using calibrated columns containing various molecular sizing matrices gave values of 1395 +/- 130 daltons for the molecular weight of FeMoco. The measured MW of FeMoco is about twice the value expected from the minimal stoichiometry, suggesting that FeMoco may exist as Mo2Fe12S12 in NMF and DMF solutions. The charge of FeMoco in its EPR silent state was determined to be 2- per Mo by passing NMF solutions of FeMoco containing excess salts of Na+, K+, Rb+, and Mg2+ through long columns equilibrated with pure NMF and then measuring the M/Mo ratio of the emerging FeMoco. Decomposition of purified FeMoco by acid or O2-exposure followed by exhaustive methylation or silanation of the resulting mixture failed to yield any methylated or silanated homocitric acid as measured by tandem gas chromatography-mass spectrometry (GC-MS) analysis. The GC-MS procedure applied to standard homocitric acid samples and various controls readily detects methylated homocitric acid at the sub-nanomole level. We conclude that the minimum molecular formula for active oxidized (EPR silent) FeMoco in NMF and in NMF-DMF mixtures is [Mo2Fe12S12]4-, but that other small organic anions such as NMF- may be present. PMID- 8228980 TI - Differential effects of the alkylating agent N-ethoxycarbonyl-2-ethoxy-1,2 dihydroquinoline on brain alpha 2-adrenoceptors and I2-imidazoline sites in vitro and in vivo. AB - The alkylating agent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) is a peptide-coupling agent that is being used to inactivate irreversibly alpha 2 adrenoceptors and other receptors. The aim of the present study was to assess the in vitro and in vivo effects of EEDQ on the newly discovered brain I2-imidazoline sites, located mainly in mitochondria. Preincubation of rat cortical membranes with EEDQ (10(-8)-10(-5) M) markedly decreased (20-90%) the specific binding of the selective antagonist [3H]RX821002 to alpha 2-adrenoceptors without affecting that of [3H]idazoxan (in the presence of adrenaline) to I2-imidazoline sites. In EEDQ-pretreated membranes (10(-5) M, 30 min at 25 degrees C), the density of I2 imidazoline sites (Bmax = 80 +/- 4 fmol/mg of protein) was not different from that determined in untreated membranes in the presence of (10(-6) M (-) adrenaline (Bmax = 83 +/- 4 fmol/mg of protein), and both densities were lower (24%, p < 0.05) than the total native density of [3H]idazoxan binding sites (Bmax = 107 +/- 6 fmol/mg of protein) (I2-imidazoline sites plus alpha 2 adrenoceptors). Treatment of rats with an optimal dose of EEDQ (1.6 mg/kg, i.p., for 2 h to 30 days) reduced maximally at 6 h (by 95 +/- 1%) the specific binding of [3H]-RX821002 to alpha 2-adrenoceptors, but also the binding of [3H]idazoxan to I2-imidazoline sites (by 44 +/- 5%). Pretreatment with yohimbine (10 mg/kg, i.p.) fully protected against EEDQ-induced alpha 2-adrenoceptor inactivation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228981 TI - Antibodies specific for GABAA receptor alpha subunits reveal that chronic alcohol treatment down-regulates alpha-subunit expression in rat brain regions. AB - Chronic administration of ethanol results in the development of tolerance and dependence. The molecular mechanism underlying these behavioral actions of ethanol is poorly understood. Several lines of evidence have suggested that some of the pharmacological actions of ethanol are mediated via a potentiation of GABAergic transmission. Chronic ethanol administration results in a reduction in the GABAA receptor-mediated 36Cl- uptake in cortical synaptoneurosomes and primary cultured neurons. We and others have shown that it also results in a 40 50% reduction in GABAA receptor alpha-subunit mRNA levels in the rat cerebral cortex. In the present study, we investigated the expression of alpha 1, alpha 2, and alpha 3 subunits of the GABAA receptor in the cerebral cortex and the alpha 1 subunit in the cerebellum by immunoblotting using polyclonal antibodies raised against alpha 1-, alpha 2-, and alpha 3-subunit polypeptides following chronic ethanol treatment. These results reveal that chronic ethanol administration to rats results in a 61 +/- 4% reduction in level of the GABAA receptor alpha 1 subunit (51 kDa), 47 +/- 8% reduction in level of the alpha 2 subunit (53 kDa), and 30 +/- 7% reduction in level of the alpha 3 subunit (59 kDa) in the cerebral cortex and a 56 +/- 5% reduction in content of the alpha 1 subunit in the cerebellum. In summary, this ethanol-induced reduction in content of the GABAA receptor alpha subunits may underlie alterations in the GABAA receptor function and could be related to cellular adaptation to the functional disturbance caused by ethanol. PMID- 8228982 TI - Interacting presynaptic kappa-opioid and GABAA receptors modulate dopamine release from rat striatal synaptosomes. AB - The presynaptic regulation of stimulated dopamine release from superfused rat striatal synaptosomes by opioids and gamma-aminobutyric acid (GABA) was studied. It was found that in addition to dopamine D2 autoreceptors, calcium-dependent K(+)-stimulated [3H]dopamine release was inhibited through activation of a homogeneous population of kappa-opioid receptors in view of the potent inhibitory effect of the kappa-selective agonist U69,593 (EC50 0.2 nM) and its antagonism by norbinaltorphimine. Neither mu- nor delta-selective receptor agonists affected release of [3H]-dopamine. In addition, GABA potently inhibited the evoked [3H]dopamine release (EC50 0.4 nM) through activation of GABAA receptors in view of the GABA-mimicking effect of muscimol, the sensitivity of its inhibitory effect to picrotoxin and bicuculline, and the absence of an effect of the GABAB receptor agonist baclofen. In the presence of a maximally effective concentration of GABA, U69,593 did not induce an additional release-inhibitory effect, indicating that these receptors and the presynaptic D2 receptor are colocalized on the striatal dopaminergic nerve terminals. The excitatory amino acid agonists N-methyl-D-aspartate and kainate, as well as the cholinergic agonist carbachol, stimulated [3H]dopamine release, which was subject to kappa-opioid receptor mediated inhibition. In conclusion, striatal dopamine release is under regulatory control of multiple excitatory and inhibitory neurotransmitter by activation of colocalized presynaptic receptors for excitatory amino acids, acetylcholine, dopamine, dynorphins, and GABA within the dopaminergic nerve terminals. Together, these receptors locally control ongoing dopamine neurotransmission. PMID- 8228983 TI - Age-related regional changes in hydroxyl radical stress and antioxidants in gerbil brain. AB - The levels of hydroxyl radicals and oxidized GSH have been examined as indices of oxidative stress in young (3 months), middle-aged (15 months), and old (20-24 months) gerbil brain hippocampus, cortex, and striatum. The hydroxyl radical stress was estimated by measuring the salicylate hydroxyl radical trapping products 2,5- and 2,3-dihydroxybenzoic acid. The stress was significantly higher in all three brain regions in middle-aged and old gerbils versus young animals (< or = 66.0%). Regional comparisons showed that the stress was significantly higher in cortex than in either the hippocampus or striatum of the middle-aged and old gerbils (< or = 32.0%). The ratio of oxidized to total GSH also increased progressively in middle-aged and old animals in all three brain regions (p < 0.05, < or = 41.1%), further indicating a general age-related increase in oxidative stress. Parallel to this age-related increase in oxidative stress, a significant, albeit slight (8%), decrease in neuronal number in hippocampal CA1 region was observed in both the middle-aged and old animals. Possible differences in antioxidant levels were also examined. Total GSH levels were similar across age groups (variance < 12%). However, the regional comparison showed that it was highest in striatum in all age groups. The levels of alpha-tocopherol (vitamin E) were significantly higher in the middle-aged and old animals in all three regions (< or = 70.4%). Vitamin E was highest in the hippocampus and the differences between the hippocampus and the cortex and striatum increased with age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228984 TI - Messenger RNA levels of chromogranin B, secretogranin II, and VGF in rat brain after AF64A-induced septohippocampal cholinergic lesions. AB - The mRNA levels of secretogranin II, chromogranin B, and VGF were compared in brains of control and AF64A-treated rats. This toxin induces specific lesions of the septohippocampal cholinergic pathway. As a consequence of this treatment, the chromogranin B message was elevated in the dentate gyrus granule cells of the hippocampus. In the paraventricular nucleus of the hypothalamus, a concomitant elevation of the messages of secretogranin II and corticotropin-releasing factor occurred in the parvocellular neurons, and an increase of those of secretogranin II and VGF occurred in a subgroup of magnocellular neurons. Further increases for secretogranin II were seen in the amygdaloid nuclei and the reticular thalamic nuclei and increases for chromogranin B in the temporal cortex, substantia nigra compacta, and ventral tegmental area. These results indicate that the toxin induced lesion of the cholinergic pathway innervating the hippocampus apparently leads to the stimulation of several defined groups of neurons that react with an increase in the mRNA levels of their secretory peptides. We suggest that changes in mRNA expression of these peptides are useful parameters for defining neurons under chronic stimulation. PMID- 8228985 TI - ATP-induced secretion in PC12 cells and photoaffinity labeling of receptors. AB - Secretion of catecholamines by rat PC12 cells is strongly stimulated by extracellular ATP via a P2-type purinergic receptor. ATP-induced norepinephrine release was inhibited 80% when extracellular Ca2+ was absent. Only four nucleotides, ATP, ATP gamma S, benzoylbenzoyl ATP (BzATP), and 2-methylthio-ATP, gave substantial stimulation of norepinephrine release from PC12 cells. ATP induced secretion was inhibited by Mg2+, and this inhibition was overcome by the addition of excess ATP suggesting that ATP4- was the active ligand. ATP-induced secretion of catecholamine release was enhanced by treatment of cells with pertussis toxin or 12-O-tetradecanoylphorbol 13-acetate. The stimulatory effects of 12-O-tetradecanoylphorbol 13-acetate and pertussis toxin on norepinephrine release were additive. After brief exposure of intact cells to the photoaffinity analog, [alpha-32P]BzATP, two major proteins of 44 and 50 kDa and a minor protein of 97 kDa were labeled. An excess of ATP gamma S and BzATP but not GTP blocked labeling of the proteins by [32P]BzATP. Labeling of the 50-kDa protein was more sensitive to competition by 2-methylthio-ATP than the other labeled proteins, suggesting that the 50-kDa protein represents the P2 receptor responsible for ATP stimulated secretion in these cells. PMID- 8228986 TI - Maintenance of neuronal glutathione by glial cells. AB - Glutathione levels in neurons and glial cells were investigated in a neuronal glial coculture and in separate cultures. Brain cell suspensions obtained from cerebral hemispheres of fetal rats were cultured, and after 5 days the glutathione content of this cell population, consisting mainly of neurons and astroglial cells, was 23.0 nmol/mg of cell protein, with a significantly high content in glial cells (28.0 nmol/mg of protein) in comparison with neurons (18.8 nmol/mg of protein). When the neurons and glial cells were separated and recultured in fresh medium, neuronal glutathione rapidly decreased, whereas glial glutathione remained unchanged. Cysteine is a rate-limiting precursor for glutathione synthesis, and its level was also decreased in neurons, but not in glial cells. Cysteine was taken up rapidly by both neurons and glial cells, but cystine was taken up only by glial cells. This accounts for the rapid decrease of glutathione in the cultured neurons, because the culture medium contains cystine, but not cysteine. It was also found that the cultured glial cells released cysteine into the medium. These results suggest that neurons maintain their glutathione level by taking up cysteine provided by glial cells. PMID- 8228987 TI - Coupling among energy failure, loss of ion homeostasis, and phospholipase A2 and C activation during ischemia. AB - The objective of the present experiments was to correlate changes in cellular energy metabolism, dissipative ion fluxes, and lipolysis during the first 90 s of ischemia and, hence, to establish whether phospholipase A2 or phospholipase C is responsible for the early accumulation of phospholipid hydrolysis products. Ischemia was induced for 15-90 s in rats, extracellular K+ (K+e) was recorded, and neocortex was frozen in situ for measurements of labile tissue metabolites, free fatty acids, and diacylglycerides. Ischemia of 15- and 30-s duration gave rise to a decrease in phosphocreatine concentration and a decline in the ATP/free ADP ratio. Although these changes were accompanied by an activation of K+ conductances, there were no changes in free fatty acids until after 60 s, when free arachidonic acid accumulated. An increase in other free fatty acids and in total diacylglceride content did not occur until after anoxic depolarization. The results demonstrate that the early functional changes, such as activation of K+ conductances, are unrelated to changes in lipids or lipid mediators. They furthermore suggest that the initial lipolysis occurs via both phospholipase A2 and phospholipase C, which are activated when membrane depolarization leads to influx of calcium into cells. PMID- 8228988 TI - Lithium modulation of phosphoinositide signaling system in rat cortex: selective effect on phorbol ester binding. AB - Recent work indicates that the therapeutic action of lithium may be mediated through perturbation of postreceptor second messenger systems. To elucidate further the postreceptor cellular sites of action(s) of lithium, the effect of chronic lithium treatment on various components of the receptor-activated phosphoinositide pathway was investigated. We found that chronic administration of lithium (0.2% LiCl, 21 days) to adult male rats did not significantly affect phosphoinositide hydrolysis in cerebral cortical slices induced by carbachol (1 mM) or NaF (10 mM). Nor did the same treatment alter the carbachol (1 mM) potentiation of guanosine 5'-(gamma-thio)triphosphate (30 microM) stimulation of phosphoinositide hydrolysis (an index of receptor/G protein coupling) in cortical membranes. Immunoblotting studies revealed no changes in the levels of G alpha q/11 immunoreactivity in the cortex after chronic lithium treatment. The levels of protein kinase C, as revealed by specific binding of [3H]phorbol dibutyrate ([3H]PDBu), were significantly reduced in the cytosolic fraction and increased in the particulate fraction of rat cortex after chronic lithium, whereas the KD of [3H]PDBu binding remained relatively constant. A small and insignificant decrease in the density of [3H]inositol 1,4,5-trisphosphate binding was also found in the cortex. The above data suggest that chronic lithium treatment affects neither the muscarinic cholinergic-linked phosphoinositide turnover nor the putative G protein alpha subunit (G alpha q/11) responsible for phospholipase C activation. However, a possible translocation and activation of protein kinase C activity may be significant in the therapeutic effect of this mood-stabilizing agent. PMID- 8228989 TI - High activities of glutamine transaminase K (dichlorovinylcysteine beta-lyase) and omega-amidase in the choroid plexus of rat brain. AB - Certain halogenated hydrocarbons, e.g., dichloroacetylene, are nephrotoxic to experimental animals and neurotoxic to humans; cysteine-S-conjugate beta-lyases may play a role in the nephrotoxicity. We now show that with dichlorovinylcysteine as substrate the only detectable cysteine-S-conjugate beta lyase in rat brain homogenates is identical to glutamine transaminase K. The predominant (mitochondrial) form of glutamine transaminase K in rat brain was shown to be immunologically distinct from the predominant (cytosolic) form of the enzyme in rat kidney. Glutamine transaminase K and omega-amidase (constituents of the glutaminase II pathway) activities were shown to be widespread throughout the rat brain. However, the highest specific activities of these enzymes were found in the choroid plexus. The high activity of glutamine transaminase K in choroid plexus was also demonstrated by means of an immunohistochemical staining procedure. Glutamine transaminase K has a broad specificity toward amino acid and alpha-keto acid substrates. The omega-amidase also has a broad specificity; presumably, however, the natural substrates are alpha-ketoglutaramate and alpha ketosuccinamate, the alpha-keto acid analogues of glutamine and asparagine, respectively. The high activities of both glutamine transaminase K and omega amidase in the choroid plexus suggest that the two enzymes are linked metabolically and perhaps are coordinately expressed in that organ. The data suggest that the natural substrate of glutamine transaminase K in rat brain is indeed glutamine and that the metabolism of glutamine through the glutaminase II pathway (i.e., L-glutamine and alpha-keto acid-->alpha-ketoglutarate and L-amino acid + ammonia) is an important function of the choroid plexus. Moreover, the present findings also suggest that any explanation of the neurotoxicity of halogenated xenobiotics must take into account the role of glutamine transminase K and its presence in the choroid plexus. PMID- 8228990 TI - Protein kinase FA/GSK-3 phosphorylates tau on Ser235-Pro and Ser404-Pro that are abnormally phosphorylated in Alzheimer's disease brain. AB - Previously, we identified protein kinase FA/glycogen synthase kinase-3 (GSK-3) as a microtubule-associated protein tau kinase that can incorporate 4 mol of phosphates into 1 mol of tau protein and cause its electrophoretic mobility shift in sodium dodecyl sulfate gels, a unique property characteristic of paired helical filament-associated pathological tau (PHF-tau) in Alzheimer's disease brains. In this report, we identified TPPKS(p)PSAAK and SPVVSGDTS(p)PR as two phosphorylation site sequences phosphorylated by kinase FA/GSK-3 in tau using peptide sequence analysis and sequential manual Edman degradation for radiosequencing. When mapping with human brain tau sequence, we further identified Ser235-Pro and Ser404-Pro as the two major phosphorylation sites according to the numbering of the longest tau isoform. Ser235 and Ser404 have been reported as two of the major abnormal phosphorylation sites in PHF-tau. Taken together, the results provide initial evidence that protein kinase FA/GSK-3 may represent one of the Ser-Pro motif-directed tau kinases involved in the abnormal phosphorylation of pathological PHF-tau in Alzheimer's disease brain. PMID- 8228991 TI - Role of the cellular stress response in the biogenesis of cysteamine-induced astrocytic inclusions in primary culture. AB - Cysteamine (CSH; 2-mercaptoethylamine) stimulates the accumulation of peroxidase positive inclusions in cultured astroglia akin to those observed in the aging periventricular brain. Because CSH induces the synthesis of a stress protein (heme oxygenase) in rat liver, we hypothesized that aspects of the cellular stress response may play a role in the biogenesis of CSH-induced astrocyte granules. In the present study, we performed indirect immunofluorescent staining and immunoblotting for various stress proteins in rat neuroglial cultures. Exposure of astrocyte cultures to CSH enhanced immunostaining for heme oxygenase 1 (HO-1) and heat-shock proteins 27, 72, and 90, but not glucose-regulated protein 94, relative to untreated cultures. CSH-pretreated astrocytes exhibited enhanced tolerance to H2O2 toxicity relative to untreated cells, providing physiological evidence of an antecedent stress response in the former. In addition, exposure for 12 days to H2O2, a known inducer of the stress response, elicited astrocyte granulation similar to that observed with CSH. Chronic induction of HO-1 and other stress proteins may participate in the biogenesis of metalloporphyrin-rich inclusions in CSH-treated astroglial cultures and in astrocytes of the aging periventricular brain. PMID- 8228992 TI - Glial fibrillary acidic protein and the mesolimbic dopamine system: regulation by chronic morphine and Lewis-Fischer strain differences in the rat ventral tegmental area. AB - In this study we demonstrate that a 51-kDa phosphoprotein, previously identified as morphine regulated and showing different basal levels among rat strains, is glial fibrillary acidic protein (GFAP). Chronic morphine increased levels of GFAP immunoreactivity by > 70% in the ventral tegmental area (VTA) of outbred Sprague Dawley rats. This increase in GFAP content was not observed in rats that were treated concomitantly with morphine and naltrexone, an opiate receptor antagonist, and did not occur in response to a single acute injection with morphine. No alterations in GFAP levels were observed in response to chronic morphine in several other regions of the CNS studied, including the substantia nigra, locus coeruleus, cerebral cortex, and spinal cord. There were also inherent differences in levels of GFAP immunoreactivity in the VTA of drug-naive Fischer 344 and Lewis rats, two inbred rat strains that differ in their relative preference for morphine and other drugs of abuse. The VTA of drug-naive Lewis rats contained more than twofold higher levels of GFAP compared with drug-naive Fischer rats. This strain difference was also apparent in the locus coeruleus but not in several other brain regions or in spinal cord. Because the mesolimbic dopamine system is thought to play a critical role in mediating the reinforcing properties of opiates and other drugs of abuse, it is possible that the opiate induction of GFAP and inherent Lewis versus Fischer strain differences in GFAP levels in the VTA may be related to the reinforcing and/or addictive properties of opiates mediated by this brain region, as well as to genetic differences in drug preference. PMID- 8228993 TI - Phosphorylation of rat brain choline acetyltransferase and its relationship to enzyme activity. AB - Choline acetyltransferase catalyzes the formation of acetylcholine from choline and acetyl-CoA in cholinergic neurons. The present study examined conditions for modulation of kinase-mediated phosphorylation of this enzyme. By using a monospecific polyclonal rabbit anti-human choline acetyltransferase antibody to immunoprecipitate cytosolic and membrane-associated subcellular pools of enzyme from rat hippocampal synaptosomes, we determined that only the cytosolic fraction of the enzyme (67,000 +/- 730 daltons) was phosphorylated under basal, unstimulated conditions. The quantity of this endogenous phosphoprotein was dependent, in part, upon the level of intracellular calcium, with 32Pi incorporation into the enzyme in nerve terminals incubated in nominally calcium free medium only 43 +/- 7% of control. The corresponding enzymatic activity of cytosolic choline acetyltransferase did not appear to be altered by lowered cytosolic calcium, whereas membrane-associated choline acetyltransferase activity was decreased to 58 +/- 11% of control. Depolarization of synaptosomes with 50 microM veratridine neither altered the extent of phosphorylation or specific activity of cytosolic choline acetyltransferase, nor induced detectable phosphorylation of membrane-associated choline acetyltransferase, although the specific activity of the membrane-associated enzyme was increased to 132 +/- 5% of control. In summary, phosphorylation of choline acetyltransferase does not appear to regulate cholinergic neurotransmission by a direct action on catalytic activity of the enzyme. PMID- 8228994 TI - Extracellular ATP stimulates calcium influx in neuroblastoma x glioma hybrid NG108-15 cells. AB - ATP-induced changes in the intracellular Ca2+ concentration ([Ca2+]i) in neuroblastoma x glioma hybrid NG108-15 cells were studied. Using the fluorescent Ca2+ indicator fura-2, we have shown that the [Ca2+]i increased in response to ATP. ATP at 3 mM caused the greatest increased in [Ca2+]i, whereas at higher concentrations of ATP the response became smaller. Two nonhydrolyzable ATP analogues, adenosine 5'-thiotriphosphate and 5'-adenylyl-beta, gamma imidodiphosphate, could not trigger significant [Ca2+]i change, but they could block the ATP effect. Other adenine nucleotides, including ADP, AMP, alpha beta methylene-ATP, beta, gamma-methylene-ATP, and 2-methylthio-ATP, as well as UTP and adenosine, all had no effect on [Ca2+]i at 3 mM. In the absence of extracellular Ca2+, the effect of ATP was inhibited totally, but could be restored by the addition of Ca2+ to the cells. Upon removal of Mg2+, the maximum increase in [Ca2+]i induced by ATP was enhanced by about 42%. Ca(2+)-channel blockers partially inhibited the ATP-induced [Ca2+]i rise. The ATP-induced [Ca2+]i rise was not affected by thapsigargin pretreatment, though such pretreatment blocked bradykinin-induced [Ca2+]i rise completely. No heterologous desensitization of [Ca2+]i rise was observed between ATP and bradykinin. The magnitude of the [Ca2+]i rise induced by ATP increased between 1.5 and 3.1 times when external Na+ was replaced with Tris, N-methyl-D-glucamine, choline, or Li+. The addition of EGTA or verapamil to cells after their maximum response to ATP immediately lowered the [Ca2+]i to the basal level in Na(+)-containing or Na(+) free Tris solution.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8228995 TI - Depression of neuronal protein synthesis initiation by protein tyrosine kinase inhibitors. AB - Growth factors stimulate cellular protein synthesis, but the intracellular signaling mechanisms that regulate initiation of mRNA translation in neurons have not been clarified. A rate-limiting step in the initiation of protein synthesis is the formation of the ternary complex among GTP, eukaryotic initiation factor 2 (eIF-2), and the initiator tRNA. Here we report that genistein, a specific tyrosine kinase inhibitor, decreases tyrosine kinase activity and the content of phosphotyrosine proteins in cultured primary cortical neurons. Genistein inhibits protein synthesis by > 80% in a dose-dependent manner (10-80 micrograms/ml) and concurrently decreases ternary complex formation by 60%. At the doses investigated, genistein depresses tyrosine kinase activity and concomitantly stimulates PKC activity. We propose that a protein tyrosine kinase participates in the initiation of protein synthesis in neurons, by affecting the activity of eIF-2 directly or through a protein kinase cascade. PMID- 8228996 TI - Detection of tau proteins in normal and Alzheimer's disease cerebrospinal fluid with a sensitive sandwich enzyme-linked immunosorbent assay. AB - Alzheimer's disease is a progressive degenerative dementia characterized by the abundant presence of neurofibrillary tangles in neurons. This study was designed to test whether the microtubule-associated protein tau, a major component of neurofibrillary tangles, could be detected in CSF. Additionally, we investigated whether CSF tau levels were abnormal in Alzheimer's disease as compared with a large group of control patients. We developed a sensitive sandwich enzyme-linked immunosorbent assay using AT120, a monoclonal antibody directed to human tau, as a capturing antibody. With this technique, the detection limit for tau was less than 5 pg/ml of CSF. Using AT8, which recognizes abnormally phosphorylated serines 199-202 in tau, the detection limit was below 20 pg/ml of CSF. However, with AT8, we found no immunoreactivity in CSF, suggesting that only a small fraction of CSF tau contains the abnormally phosphorylated AT8 epitope. Our results indicate that CSF tau levels are significantly increased in Alzheimer's disease. Also, CSF tau levels in a large group of patients with a diversity of neurological diseases showed overlap with CSF tau levels in Alzheimer's disease. PMID- 8228997 TI - Free fatty acid and diacylglycerol accumulation in the rat brain during recurrent seizures is related to cortical oxygenation. AB - Cerebral blood flow and oxygenation increase during the early seizures of a series, but the increase in cerebral blood flow attenuates during late seizures, sometimes resulting in decreased cortical oxygenation. Cortical free fatty acids (FFA) and diacylglycerols also increase during early seizures and the increase attenuates during late seizures. We analyzed the correlation between lipid accumulation and cortical O2 during periodic pentylenetetrazol-induced seizures. During early seizures, both FFA and diacylglycerols increased in the cerebral cortex, particularly arachidonate (20:4) and stearate (18:0). Changes in lipids were different during late seizures, depending on cortical O2 levels. An increase in cortical O2 during late seizures was associated with lower FFA levels compared with early seizures, and FFA levels recovered to basal levels during interictal periods. A decline in cortical O2 was associated with a further increase in FFA, which remained elevated during interictal periods. Our results indicate that periseizure lipid accumulation is related to cortical oxygenation. PMID- 8228998 TI - Altered composition of cerebral microvessel membrane phosphoglycerides from senescent mouse. AB - Phosphoglyceride and fatty acid composition was determined in the cellular membranes of isolated cerebral microvessels and brain parenchymal cells (neurons and glia) taken from 10-, 20-, and 27-30-month-old C57BL6/NNIA mice. Lipids were extracted from each fraction and the fatty acid profiles of ethanolamine, choline, serine, and inositol phosphoglycerides analyzed by gas chromatography. The results suggest that membrane phosphoglycerides from cerebral microvessels are significantly more affected by the aging process than are those of the brain parenchyma. Relative percentage for fatty acids in cerebral microvessels indicate an overall decline in membrane unsaturation with a concomitant elevation in the level of saturation. The decline in unsaturation is reflected primarily in the loss of precursor fatty acids for arachidonic (18:2n-6 and 20:3n-6) and docosahexaenoic (20:5n-3 and 22:5n-3) acids. Levels of arachidonic (20:4n-6) and docosahexaenoic (22:6n-3) acids in each phosphoglyceride remained unchanged with age; however, mol% for ethanolamine plasmalogen, a major source of these fatty acids, was significantly reduced in 27-30-month-old mice. Conversely, mol% for choline phosphoglyceride increased with age. The age-related changes in fatty acid profile for microvessel membrane phosphoglycerides are reflected by increased saturation/unsaturation ratios and decreased unsaturation indices. These parameters were not affected by aging in parenchymal membranes. PMID- 8228999 TI - Influence of potassium concentration in microdialysis perfusate on basal and stimulated striatal dopamine release: effect of ceruletide, a cholecystokinin related peptide. AB - The in vivo microdialysis method was used to study the effect of the cholecystokinin-related peptide, ceruletide, on extracellular levels of dopamine (DA) in the striatum following perfusion with various K+ concentrations. Increasing the K+ concentration in the perfusate from 4 to 15 or 17.5 mM did not change basal DA release or release evoked by electrical stimulation of the medial forebrain bundle (MFB). However, when the perfusing solution contained 20 or 30 mM K+, dose-dependent reductions of both basal and MFB-stimulated DA release occurred. Subcutaneous administration of ceruletide at 160 micrograms/kg had no influence on the basal or MFB-stimulated DA release with 4 or 15 mM K+ in the perfusate. However, after perfusion with 17.5 mM K+, ceruletide significantly attenuated the basal and MFB-stimulated DA release. Carbachol (10 microM) locally applied via the dialysis probe also attenuated MFB-stimulated DA release after perfusion with 17.5 mM K+. From these results, we conclude that under appropriate depolarization of striatal DA terminals, ceruletide induces further depolarization and inactivation of nigrostriatal DA terminals. The present data suggest that this effect may be mediated via intrinsic cholinergic neurons in the striatum. PMID- 8229000 TI - A possible pathway of phosphoinositide metabolism through EDTA-insensitive phospholipase A1 followed by lysophosphoinositide-specific phospholipase C in rat brain. AB - Incubation of [2-3H]glycerol-labeled phosphatidylinositol with a crude cytosol fraction of rat brain in the presence of EDTA yielded [3H]lysophosphatidylinositol predominantly without accumulation of labeled monoacylglycerol and diacylglycerol. The pH optimum of this phospholipase A activity was 8.0. The activity for phosphatidylinositol was twofold higher than for phosphatidylethanolamine, whereas phosphatidylcholine, phosphatidylserine, and phosphatidic acid were not hydrolyzed significantly under the conditions used. The phospholipase A activity for phosphatidylethanolamine was resolved in part from that for phosphatidylinositol by ammonium sulfate fractionation of the cytosol, indicating the existence of at least two forms of EDTA-insensitive phospholipase A. The positional specificity of the phosphatidylinositol hydrolyzing activity was found to be that of a phospholipase A1, as radioactive lysophosphatidylinositol was produced from 1-stearoyl-2-[1-14C]arachidonyl-sn glycero-3-phosphoinositol without release of free arachidonate. A phospholipase C activity specific for lysophosphoinositides was found in a membrane fraction from rat brain, which was similar to that characterized in porcine platelets. The phospholipase C was demonstrated to hydrolyze the 2-acyl isomer as well as the 1 acyl isomer of lysophosphatidylinositol. Taken together, our results suggest a possible pathway through which phosphatidylinositol is selectively degraded to the 2-acyl isomer of lysophosphatidylinositol in a Ca(2+)-independent manner, and subsequently converted to a 2-monoacylglycerol in rat brain. PMID- 8229001 TI - Prenatal ethanol exposure: changes in regional brain catecholamine content following stress. AB - Previous studies have shown that fetal ethanol exposure (FEE) may have long-term effects on the function of catecholaminergic neurons in different regions of the CNS. The present study is the first to examine the effects of FEE on regional brain catecholamine responses following acute stress (a single 60-min restraint stress), repeated stress (single periods of restraint stress on 1, 5, or 10 consecutive days), and recovery from stress (recovery for up to 60 min in the home cage following a single 60-min period of restraint stress). Both male and female offspring from FEE, pair-fed (PF), and ad libitum-fed control (C) groups were tested in adulthood to determine catecholamine content in the cortex, hypothalamus, and hippocampus. A single period of restraint reduced cortical norepinephrine (NE) content in FEE and PF animals compared with that in the cortex of C animals, and reduced hypothalamic NE content in FEE female offspring below that found in animals in all other groups. In contrast, hippocampal NE content was higher in FEE than in C animals following a single period of restraint; PF animals had intermediate levels of hippocampal NE and did not differ significantly from either FEE or C animals. Following repeated periods of restraint, cortical NE content was lower in FEE than in C animals; PF animals once again had intermediate levels of NE. Importantly, basal (nonstressed) NE content did not differ among groups in any brain area examined. In addition, several significant changes in regional brain catecholaminergic responses to acute stress were observed in animals across all treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229002 TI - Morphological and biochemical analyses of amyloid plaque core proteins purified from Alzheimer disease brain tissue. AB - Amyloid plaque cores were purified from Alzheimer disease brain tissue. Plaque core proteins were solubilized in formic acid which upon dialysis against guanidinium hydrochloride (GuHCl) partitioned into soluble (approximately 15%) and insoluble (approximately 85%) components. The GuHCl-soluble fraction contained beta-amyloid1-40, whereas the GuHCl-insoluble fraction was fractionated into six components by size exclusion HPLC: S1 (> 200 kDa), S2 (200 kDa), S3 (45 kDa), S4 (15 kDa), S5 (10 kDa), and S6 (5 kDa). Removal of the GuHCl reconstituted 10-nm filaments composed of two intertwined 5-nm strands. Fractions S5 and S6 also yielded filamentous structures when treated similarly, whereas fractions S1-S4 yielded amorphous aggregates. Chemical analysis identified S4-S6 as multimeric and monomeric beta-amyloid. Immunochemical analyses revealed alpha 1-antichymotrypsin and non-beta-amyloid segments of the beta-amyloid precursor protein within fractions S1 and S2. Several saccharide components were identified within plaque core protein preparations by fluorescence and electron microscopy, as seen with fluorescein isothiocyanate- and colloidal gold-conjugated lectins. We have shown previously that this plaque core protein complex is more toxic to neuronal cultures than beta-amyloid. The non-beta-amyloid components likely mediate this additional toxicity, imposing a significant influence on the pathophysiology of Alzheimer disease. PMID- 8229003 TI - Pharmacological characterization of 5-hydroxytryptamine3 receptors in murine brain and ileum using the novel radioligand [3H]RS-42358-197: evidence for receptor heterogeneity. AB - Previous studies have demonstrated species-specific differences in 5 hydroxytryptamine3 (5-HT3) receptors, but unequivocal evidence of 5-HT3 receptor subtypes, within a species, has not yet been obtained. The purpose of the current study was to test for heterogeneity in 5-HT3 receptors in murine tissues. 5-HT3 receptors in membranes derived from brain cerebral cortex of CD-1, C57Bl/6, and Swiss Webster mice and ileum of CD-1 mice were labeled with the 5-HT3 receptor antagonist [3H]RS-42358-197. Structurally diverse competing ligands were then used to characterize the binding site. [3H]RS-42358-197 bound with similar affinity in each of the cortical tissues (mean KD = 0.14 nM; range, 0.06-0.32 nM) but bound with lower affinity in ileal tissue (2.5 nM). The density of sites labeled with [3H]RS-42358-197 ranged from 10.4 fmol/mg of protein in Swiss Webster mouse cortex to 44.2 fmol/mg of protein in Sprague-Dawley rat cortex. Displacing ligands produced a pharmacologic profile of the [3H]RS-42358-197 binding site consistent with it being a 5-HT3 receptor: (R)-YM060 > (S)-zacopride > (R)-zacopride > MDL 72222 > 2-methyl-5-HT. However, > or = 10-fold differences in the affinity of certain ligands were found when comparing 5-HT3 binding sites in membranes from cerebral cortex of the different strains of mice and when comparing 5-HT3 binding sites in brain and ileal membranes prepared from the CD-1 mouse strain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229004 TI - Characterization of beta-amyloid peptide from human cerebrospinal fluid. AB - beta-Amyloid peptide (A beta) is one of the main components of senile plaques in the brain tissue of Alzheimer's disease (AD) patients. A beta is proteolytically cleaved from the amyloid precursor protein (APP), an integral membrane protein possessing a large extracellular N-terminal domain followed by a single membrane spanning region and a short cytoplasmic C-terminal tail. A beta has been isolated from senile plaques and cerebral vascular tissue of AD brain and characterized as a heterogeneous peptide containing 28-43 amino acids whose sequence begins in the extracellular domain of APP and extends into the putative transmembrane sequence. It has long been speculated that A beta may also be present in body fluids, such as CSF, that contact neuritic plaques. Recently using a specific enzyme-linked immunosorbent assay we were able to quantify one form of A beta in CSF. In this report, using one of these antibodies covalently bound as an affinity matrix, multiple complex forms of A beta have been isolated and characterized from CSF derived from patients with either meningitis or other neurological disorders. Amino acid sequencing reveals A beta species with N-termini of Asp1, Glu3, His6, Glu11, and Val12, although on a molar basis, Asp1 represents the predominant aminoterminus. Laser desorption mass spectrometry confirmed the presence in CSF of A beta species containing 27, 28, 30, 34, 35, 40, 42, and 43 amino acids, all beginning at Asp1; two stable trimers, (Asp1-Met35)3 and (His6-Ala42)3; and one stable dimer containing (Asp1-Val40)2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229005 TI - GM3 regulates protein kinase systems in cultured brain microvascular endothelial cells. AB - The barrier function of endothelial cells is known to be positively regulated by protein kinase A (PKA) and negatively regulated by protein kinase C (PKC). We found that exogenously administered GM3(NeuAc) promoted PKA activity in cultured brain microvascular endothelial cells (BMECs). Other glycolipids, including GM1, sulfoglucuronyl paragloboside, and GM3(NeuGc), did not have any effect on the PKA activity of BMECs. PC12 cells did not respond to exogenously applied GM3(NeuAc). GM3(NeuAc) also suppressed the PKC activity of BMECs. Thus, GM3(NeuAc) may function as a modulator of blood-brain barrier function via the two different kinase systems. PMID- 8229006 TI - Regional variability in DNA fragmentation after global ischemia evidenced by combined histological and gel electrophoresis observations in the rat brain. AB - We have studied whether the delayed cell death induced by transient forebrain ischemia is associated with an internucleosomal cleavage of DNA into oligonucleosome-sized fragments. The integrity of genomic DNA in various brain regions after a 20-min four-vessel ischemia was examined using gel electrophoresis. We found typical ladders of oligonucleosomal DNA fragments in the striatum and in the Ammon's horn. In the latter we also often found a random DNA degradation as a smear pattern. These findings were reinforced by a specific in situ labeling of DNA breaks in tissue sections. A dark staining of nuclei was observed in the cell bodies of neurons--in particular in the head of the caudate and in the vulnerable CA1 hippocampal area. With biochemical and histological approaches, there was no evidence of DNA degradation in regions that are resistant to the injury. We conclude that the association of multiple mechanisms of cell damage may occur after a global ischemia. The regional variability in DNA fragmentation stresses the importance of using histological approaches in parallel with gel electrophoresis. PMID- 8229007 TI - ras-independent induction of rat brain type II sodium channel expression in nerve growth factor-treated PC12 cells. AB - Nerve growth factor (NGF) plays an important role in the development of the nervous system, and there is considerable interest in understanding the molecular mechanisms underlying its effects on neuronal differentiation. To determine if the activity of proteins of the ras gene family is necessary for the NGF-mediated induction of sodium channel expression in pheochromocytoma (PC12) cells, sodium channel expression was analyzed in PC12 sublines stably overexpressing the dominant inhibitory mutant c-Ha-ras(Asn-17). Northern blot analysis, RNase protection assays, and whole-cell patch clamp recordings indicate that the NGF mediated increase in type II sodium channel mRNA and sodium current density can occur independent of ras activity and by doing so provide strong evidence for the importance of ras-independent mechanisms in NGF-mediated neuronal differentiation. PMID- 8229008 TI - Differential enhancement of benzophenanthridine alkaloid content in cell suspension cultures of Sanguinaria canadensis under conditions of combined hormonal deprivation and fungal elicitation. AB - An elicitation protocol, resulting in the accumulation of sanguinarine in suspension cultures of Papaver bracteatum, was assessed for induction of the same alkaloid in Sanguinaria canadensis. Although only a trace constituent of P. bracteatum plants, sanguinarine is a major alkaloid (1-3% dry wt) of S. canadensis rhizomes. By combining hormonal deprivation for various intervals and a 3-day fungal (Verticillium dahliae) elicitation, benzophenanthridine alkaloid accumulation was induced in S. canadensis cell suspensions. Chelirubine content increased (0.1-1.3% dry wt) consistently in elicited cell cultures while chelerythrine (0.01-0.10% dry wt) and sanguinarine (0-0.02% dry wt) levels were considerably less. Alkaloid accumulation always occurred upon removal of hormone but only at certain time intervals in the log phase upon fungal elicitation. Levels of dopamine, a precursor of the alkaloids, fluctuated over the incubation period, but displayed a 2- to 6-fold increase in cell suspensions grown without hormone. In some experiments dopamine accumulated to levels > 20% dry wt, and these increases were enhanced by the addition of fungal elicitor. Although the same fungal elicitor induces benzophenanthridines in taxonomically related S. canadensis and P. bracteatum, it did not elicit the accumulation of the same alkaloid in the two different plant cultures. PMID- 8229009 TI - New cyanocyclines from a cyanide-treated broth of Streptomyces lusitanus. AB - Organic extracts of Streptomyces lusitanus, the producer of the anticancer antibiotic naphthyridinomycin [1], were found to contain two additional compounds active in an antibiotic screen. As with 1, these reacted with NaCN added at the end of the fermentation. One of the addition products has been named cyanocycline B [3] and is derived from N-desmethylnaphthyridinomycin [4], while the other has been named cyanocycline C [5], and is derived from the hydroquinone 6 of 1. Cyanocycline C is unstable and was characterized after conversion to the dimethyl derivative with CH2N2 in the presence of TFA. The implications of these metabolites for the biosynthesis of 1 are discussed. A third new antibiotic, cyanocycline D [8] was isolated from the cyanide-treated broth and proved to be an artifact in which the oxazolidine ring had been opened by cyanide. The potential relevance of the formation of 8 to the reaction of 1 with DNA is also discussed. PMID- 8229010 TI - The identification of 5-hydroxy-L-norvaline in cultures of pyridoxine auxotrophs of Escherichia coli B. AB - Gc-ms and hplc have been employed to identify 5-hydroxy-L-norvaline (L-2-amino-5 hydroxypentanoic acid) in the culture fluids of pyridoxine-starved cultures of two pyridoxine auxotrophs of Escherichia coli B. The production of 5 hydroxynorvaline is shown to be secondary to pyridoxine starvation of the cultures, and the quantities of 5-hydroxynorvaline are similar to those of the common protein amino acids present in these cultures. This is the first time that 5-hydroxynorvaline has been identified as a product of bacterial metabolism. PMID- 8229011 TI - Structure of fischerin, a new toxic metabolite from an ascomycete, Neosartorya fischeri var. fischeri. AB - A new toxic metabolite named fischerin [1] from an Ascomycete. Neosartorya fischeri var. fischeri, which caused lethal peritonitis in mice, was deduced to have a structure including a 1,4-dihydroxy-3,5-disubstituted-2(1H)-pyridone moiety by chemical and spectral data. PMID- 8229012 TI - New sesquiterpene alpha-methylene lactones from the Egyptian plant Jasonia candicans. AB - Extracts of the plant Jasonia candicans possess antimicrobial activity. ET2O/MeOH extracts were subjected to liquid chromatography, and several sesquiterpenes were isolated and identified including the known compounds confertin [1], 4,11(13) eudesmadien-12-oic acid [2], and 11-eudesmen-4-ol [3]. Two new diol alpha methylene lactone antimicrobial agents were identified from nmr and mass spectral data and X-ray crystallography as (4 alpha, 5 alpha, 8 beta, 10 beta)-4,10 dihydroxy-1,11(13)-guaidien-12,8-olide [4] and (4 alpha, 5 alpha, 8 beta, 10 alpha)-4,10-dihydroxyl-1,11 (13)-guaidien-12,8-olide[5], which differ in stereochemistry at the C-10 tertiary alcohol center. PMID- 8229013 TI - Isolation and structural elucidation of tetracenomycin F2 and tetracenomycin F1: early intermediates in the biosynthesis of tetracenomycin C in Streptomyces glaucescens. AB - This report describes the fermentation, isolation, and structural elucidation of tetracenomycin (Tcm) F2 [2], a metabolite produced by a blocked mutant strain WMH1092 of the Tcm C [1] producer Streptomyces glaucescens and by the recombinant strain S. glaucescens WMH1077 (pWHM722). Elucidation of the Tcm F2 structure shows that 2 is the earliest intermediate identified to date in the biosynthesis of 1. This is supported by the fact that 2 is very efficiently biotransformed to 1 by the S. glaucescens WMH1068 strain and is enzymatically converted to Tcm F1 [3] and to Tcm D3 [4], a known intermediate of Tcm C biosynthesis. PMID- 8229014 TI - Properties of isochorismate hydroxymutase from Flavobacterium K3-15. AB - Isochorismate hydroxymutase (isochorismate synthase, E.C. 5.4.99.6) catalyzes the interconversion of chorismic acid [1] and isochorismic acid [2]. The enzyme was extracted from a Flavobacterium K3-15 that overproduces vitamin K2 (i.e., menaquinones) and was purified to homogeneity. The N-terminal amino acid sequence and the mol wt (36,240 +/- 100 daltons) were determined by ms following SDS PAG electrophoresis. The enzyme was characterized (stability, cofactor requirement, isoelectrical point), and antibodies were raised which showed no cross reactivity with isochorismate hydroxymutase from Escherichia coli and Enterobacter aerogenes 62-1. The kinetic data of the enzyme are different from those observed for the corresponding enzyme from Escherichia coli and Galium mollugo. PMID- 8229015 TI - Immobilization of isochorismate hydroxymutase. Comparison of native versus immobilized enzyme. AB - Partially purified isochorismate hydroxymutase (isochorismate synthase, E.C. 5.4.99.6) from Flavobacterium K3-15, a vitamin K overproducer, was immobilized on CNBr-activated Sepharose 4B, alkylamine glass substituted with glutardialdehyde, and aminohexyl Sepharose 4B substituted with glutardialdehyde. The immobilized enzyme exhibited a lower specific activity but a broader pH tolerance and a higher thermostability than the soluble enzyme. The stability of the enzyme was greatly increased by immobilization. Isochorismic acid, which is not commercially available, was prepared by a constant flow incubation. PMID- 8229016 TI - Cytotoxic and antimalarial alkaloids from the bulbs of Crinum amabile. AB - From the bulbs of Crinum amabile (Amaryllidaceae), a new alkaloid (-)-amabiline [1], together with the known alkaloids (-)-lycorine [2], (-)-buphanisine [3], (-) augustine [4], and (+)-crinamine [5], were isolated. The structural characterization of 1 and the revised 1H- and 13C-nmr assignments of 2 are discussed. Alkaloids 2, 4, and 5 were found to be the principal cytotoxic and antimalarial constituents. PMID- 8229017 TI - 2-Hydroxymethyl-3,4-dihydroxy-6-methyl-pyrrolidine (6-deoxy-DMDP), an alkaloid beta-mannosidase inhibitor from seeds of Angylocalyx pynaertii. AB - A polyhydroxy alkaloid has been isolated from the seeds of the African legume Angylocalyx pynaertii and identified as a 2-hydroxymethyl-3,4-dihydroxy-5 methylpyrrolidine by ms and 1H- and 13C-nmr spectroscopy. The absolute stereochemistry was established, by a stereochemically unambiguous synthesis from diacetone glucose, as 2,5-imino-1,2,5-trideoxy-D-mannitol, which may also be regarded as 2R,5R-dihydroxymethyl-3R,4R-dihydroxypyrrolidine (DMDP) [2] in which a hydroxymethyl group is deoxygenated, i.e., 6-deoxy-DMDP [1]. Whereas the structurally related polyhydroxypyrrolidine alkaloids which have previously been discovered are inhibitors of alpha- and beta-glucosidase, 6-deoxy-DMDP is unique in inhibiting beta-mannosidase. In addition to this novel alkaloid and 2 hydroxymethyl-3,4-dihydroxypyrrolidine [3], previously shown to be present in several Angylocalyx species, the known piperidine alkaloids deoxymannojirimycin [4] and fagomine [5] were identified for the first time as constituents of An. pynaertii seeds. PMID- 8229018 TI - Synthesis and reaction of potential alternate substrates and mechanism-based inhibitors of clavaminate synthase. AB - Clavaminate synthase is an FeII/alpha-ketoglutarate-dependent enzyme central to the biosynthesis of the beta-lactamase inhibitor clavulanic acid. In the presence of dioxygen it catalyzes the oxidative cyclization/desaturation of proclavaminic acid to clavaminic acid in a two-step process. Samples of (4'R)- and (4'S)-D,L [4'-2H]proclavaminic acid have been prepared and used to demonstrate that oxazolidine ring formation occurs with retention of configuration. The stereochemical course of oxygen insertion from substrate that takes place in this oxidative cyclization is the same as that observed from molecular oxygen in several hydroxylation reactions catalyzed by other FeII/alpha-ketoglutarate dependent enzymes. The ferryl (FeIV = O) species thought to be transiently involved in each of these processes was investigated in the present work with clavaminate synthase and three structural analogues of proclavaminic acid bearing vinyl or ethynyl groups at C-4' or a cyclopropyl at C-4. In the synthesis of the former two derivatives and proclavaminic acid stereoselectively labeled with deuterium at C-4', introduction of the unsaturated substituents in a stereochemically defined manner at C-4' relied upon ready access to (4R)-4 thiophenyl-2-azetidinone. Trimethylsilyl substitution could be easily achieved at C-3 of the optically pure starting material to give the readily separable cis and trans diastereomers. In radical chain reactions in which the thiophenyl was replaced by deuterium or in anionic reactions in which the thiophenyl was eliminated as its sulfone and replaced by addition of carbanions, the steric bulk of the trimethylsilyl group at C-3 governed the approach of incoming reagents to give the trans product. The enzymatic fate, however, of these derivatives was disappointing, yielding neither detectable reaction nor hoped-for inactivation of clavaminate synthase. Finally, as mixed competitive/noncompetitive inhibitors of catalysis, they gave unexceptional inhibition constants in the range 2-10 mM. PMID- 8229019 TI - Isocladosporin, a biologically active isomer of cladosporin from Cladosporium cladosporioides. AB - Extraction of the fungus Cladosporium cladosporioides yielded the known isocoumarin, cladosporin [1], and a new compound. This metabolite, which inhibited the growth of etiolated wheat coleoptiles slightly more than did cladosporin, was characterized as a diastereoisomer of cladosporin at C-14 and was named isocladosporin [2]. PMID- 8229020 TI - Cytotoxicity of Hymenocallis expansa alkaloids. AB - From the bulbs and leaves of Hymenocallis expansa (Amaryllidaceae), three alkaloid constituents were identified: (+)-tazettine, (+)-hippeastrine, and (-) haemanthidine. These alkaloids demonstrated significant cytotoxicity when tested against a panel of human and murine tumor cell lines. PMID- 8229021 TI - Inhibitory effect of carnosic acid on HIV-1 protease in cell-free assays [corrected]. AB - In order to find new effective HIV protease inhibitors, two diterpenes (carnosic acid [1] and carnosol [5]) were isolated from rosemary (Rosmarinus officinalis L.), and rosmanol [2] and semisynthetic derivatives (7-O-methylrosmanol [3], 7-O ethylrosmanol [4], and 11,12-O,O-dimethylcarnosol [6]) were prepared. The inhibitory activity of all six compounds against HIV-1 protease was tested. The carnosic acid [1] showed the strongest inhibitory effect (IC90 = 0.08 micrograms/ml). The same compound was also assayed against HIV-1 virus replication (IC90 = 0.32 micrograms/ml). The cytotoxic TC90 on H9 lymphocytes was 0.36 micrograms/ml, which is very close to the effective antiviral dose. Additionally, the tested compounds did not inhibit cellular aspartic proteases cathepsin D and pepsin at the concentration range up to 10 micrograms/ml [corrected]. PMID- 8229022 TI - Perfragilins A and B, cytotoxic isoquinolinequinones from the bryozoan Membranipora perfragilis. AB - Detailed spectroscopic and cytotoxicity data are reported for two new isoquinolinequinones, perfragilins A [1] and B [2], which were isolated from the bryozoan Membranipora perfragilis collected in South Australia. Both compounds, which are speculated to be of microbial origin, contain a relatively uncommon thiomethyl ether functionality. PMID- 8229023 TI - Threats to academic neurosurgery in the United Kingdom: a personal view. AB - Money, or lack of it, is but one, albeit very important, threat to academic neurosurgery. This review has rambled over some of the other minefields and self inflicted hurdles. The path of academic progress is long and tortuous. Time has to be found in neurosurgery to examine critically the opportunities and threats posed by a changing world to our goals of establishing standards of patient care, research, and the flexible training of the next generation. PMID- 8229024 TI - Johann Wolfgang von Goethe (1749-1832). PMID- 8229026 TI - A randomised clinical trial comparing prednisone and azathioprine in myasthenia gravis. Results of the second interim analysis. Myasthenia Gravis Clinical Study Group. AB - From January 1983 to October 1990, 41 patients with generalised myasthenia gravis were randomly given either prednisone or azathioprine. The main goal was to record the time to the occurrence of the first episode of deterioration. During a mean follow-up of 30 months, 21 patients showed deterioration, 12 in the prednisone group and nine in the azathioprine group (p = 0.40). No difference was observed between the two groups in muscular score and functional grade, assessed at the end of each treatment year, or in tolerance. Treatment failure occurred in 17 patients, 12 in the prednisone group and five in the azathioprine group (p = 0.02); even after adjustment for imbalances in prognostic features, the failure rate remained 2.8 times higher in the prednisone group than in the azathioprine group (p = 0.5). In the patients in whom treatment failed, symptoms were initially more severe than in the others, but the combination of prednisone and azathioprine resulted in clinical improvement, consisting of remission or only minor deficits in half of the patients after two years of treatment. These findings indicate that azathioprine increases treatment response compared with prednisone, although no difference in the duration of improvement was demonstrated. Nevertheless, it appears that the most severe forms of the disease, often resistant to prednisone or azathioprine alone, could benefit from the combination of both drugs. PMID- 8229025 TI - Acute behaviour disturbances. PMID- 8229027 TI - Anti-sulphatide antibodies in peripheral neuropathy. AB - A study was carried out on 135 patients with chronic idiopathic neuropathy (63), neuropathy associated with monoclonal gammopathy (51, including eight with anti MAG antibody activity) and the Guillain-Barre syndrome (GBS) (21). Serum IgM, IgG and IgA anti-sulphatide antibody titres were compared with titres in 304 patients with other neurological or immunological diseases and in 50 normal subjects. Titres were presented a) as the highest serum dilution at which reactivity could be detected, and b) in the linear region of the optical density curve. A substantial number of patients with neurological or immunological diseases had higher titres than normal subjects. Compared with normal and disease controls, five patients with neuropathy associated with IgMk monoclonal gammopathy had raised titres of IgM anti-sulphatide antibodies and one patient with GBS had raised IgM, IgG and IgA anti-sulphatide antibodies in the acute phase of the disease. Two patients had a predominantly axonal sensory neuropathy with presenting symptoms of painful paresthesiae and minimal neurological deficit. Three patients had a predominantly demyelinating sensorimotor neuropathy associated with anti-MAG antibody activity. The patient with GBS had extensive sensory loss and antibody titres returned to normal within three weeks. Raised titres of anti-sulphatide antibodies occurred in several types of neuropathy, but all had some degree of sensory impairment and associated immunological abnormality. PMID- 8229028 TI - A comparative study of simple and choice reaction time in Parkinson's, Huntington's and cerebellar disease. AB - The aim of the study was to compare the quantitative and qualitative similarities and differences in the performance of patients with Parkinson's disease, Huntington's disease and cerebellar disease on a number of reaction time tasks. Simple reaction time (SRT), uncued and fully cued four choice (CRT) tasks were performed by eight patients with Parkinson's disease after withdrawal of dopaminergic medication for an average of 14.4 hours; by seven non-demented patients with Huntington's disease and by eight patients with cerebellar disease. An S1 (warning signal/precue)-S2 (imperative stimulus) paradigm was used in all tasks, with the S1-S2 interval randomly varying between 0, 200, 800, 1600 and 3200 ms across trials. The patients with Huntington's disease had a significantly longer SRT than those with Parkinson's disease. None of the other group differences in uncued and unwarned SRT and CRT was significant. For the patients with Parkinson's disease and those with cerebellar disease, unwarned SRT was faster than uncued and unwarned CRT. For the patients with Huntington's disease, this CRT/SRT difference was not significant. A warning signal before the imperative stimulus resulted in a reduction of reaction time in all three groups. Advance information provided by S1 about the response that would be required by S2 was used by patients in all three groups, evident from reaction times in the fully cued CRT task being faster than those in the uncued CRT condition. Patients with cerebellar disease had slower movement times in the SRT and CRT conditions compared with the patients with Parkinson's disease and Huntington's disease, whose times did not differ. In one SRT condition, when the absence of a warning signal was predictable, patients with cerebellar disease, and to a lesser extent those with Huntington's disease, were able to maintain a general motor readiness before the imperative stimulus. This was not the case for the patients with Parkinson's disease who seemed more dependent on the presence of a warning signal to reduce their reaction time. With a few exceptions, the pattern of results of the three groups were qualitatively similar. It may be concluded that similar reaction time deficits are found in Parkinson's disease, in patients with other disorders of the basal ganglia (Huntington's disease), as well as those with a disease sparing the basal ganglia (cerebellar disease). The non-specific slowness observed at the behavioural level may, however, have diverse central mechanisms. PMID- 8229029 TI - Transient epileptic amnesia--a clinical update and a reformulation. AB - While absence attacks and complex partial seizures have been well documented in patients with epilepsy, the delineation of pure episodes of memory loss without additional clinical manifestations remains poorly characterised. The recently described condition of transient epileptic amnesia (TEA) is critically examined, and four new cases are described, in each of which there were episodes of pure memory loss which subsequently proved to be epileptic in origin. The anatomical and pathophysiological basis of TEA is presumed to be similar to transient global amnesia (TGA), that is, it is likely to be primarily hippocampal in origin, but with more variable involvement of limbic and adjacent temporal lobe neocortical structures. PMID- 8229030 TI - Cranial nerve palsies in spontaneous carotid artery dissection. AB - Two patients had isolated unilateral cranial nerve palsies due to spontaneous internal carotid artery (ICA) dissection without ischaemic cerebral involvement. One had acute glossopharyngeal and vagal, the other isolated hypoglossal nerve palsy. Reviewing all reported cases of angiographically confirmed ICA dissection in the literature, 36 additional cases with unequivocal ipsilateral cranial nerve palsies were analysed. While an isolated palsy of the IXth and Xth has not been reported previously, palsies of the XIIth nerve or the IXth to XIIth nerves were most frequently found. In these patients, lower cranial nerve palsies are probably the result of compression by an enlarging ICA due to mural haematoma. Symptoms and signs indicative of carotid dissection were concurrently present only in some reported cases. This raises the question of unrecognised carotid dissection as a cause of isolated cranial nerve palsies. When the dissection occurs in the subadventitial layer without relevant narrowing of the arterial lumen and when an aneurysm is thrombosed, angiography does not reliably yield the diagnosis. Therefore, carotid dissection might have been underestimated as a cause of isolated lower cranial nerve palsies before the advent of MRI. MRI demonstrates directly the extension of the wall haematoma in the axial and longitudinal planes. Some arteriopathies such as fibromuscular dysplasia and tortuosity make a vessel predisposed to dissection. PMID- 8229031 TI - Risk factors for motor neuron disease: a case-control study based on patients from the Scottish Motor Neuron Disease Register. AB - In order to identify risk factors for the subsequent development of motor neuron disease (MND) we have carried out a case-control study of incident patients in Scotland, identified using the Scottish Motor Neuron Disease Register. A standard questionnaire was given to 103 patients and the same number of community controls matched on a one to one basis using the general practitioner's (GP) age and sex register. Recall bias was minimised by using GP records to verify the subject's report. There was an overall lifetime excess of fractures in patients, odds ratio (OR) = 1.3 (95% confidence interval (CI), 0.7-2.5) and this was highest in the 5 years before symptom onset (OR = 15, 95% CI, 3.3-654). There was no association with non-fracture trauma but the OR for a manual occupation in patients was 2.6 (95% CI, 1.1-6.3). Both occupational exposure to lead (OR = 5.7, 95% CI, 1.6-30) and solvents/chemicals (OR = 3.3, 95% CI 1.3-10) were significantly more common in patients. No consistent association was found between MND and factors reflecting socioeconomic deprivation in childhood; childhood infections or social class. Our results identify a number of different factors which may contribute to the aetiology of MND. PMID- 8229032 TI - Delirium associated with Joseph disease. AB - Three Japanese patients with Joseph disease from different families developed sleep disturbance, followed by delirium at the middle to end stage. Brain CT scans of the three patients showed brainstem tegmental atrophy. EEG revealed slowing of background activity. Two necropsy cases showed degeneration of the reticular formation, raphe nuclei and locus ceruleus in the brainstem tegmentum in addition to the common pathological findings of Joseph disease. The clinicopathological correlation between the delirium and the brainstem tegmental atrophy in Joseph disease is discussed. PMID- 8229033 TI - Intermittent high-dose vitamin C therapy in patients with HTLV-I associated myelopathy. AB - The efficacy of intermittent high-dose vitamin C therapy was evaluated in seven patients with HTLV-I-associated myelopathy (HAM). All HAM patients responded well to this therapy without serious side effects. Grade of disability score improved at 9.7 (SD 5.8) months after the therapy from 7.1 (3.3) to 3.6 (2.0) (p < 0.01). Serum immunosuppressive acidic protein was elevated before and decreased after the therapy from 747 (316) to 398 (86) micrograms/ml (p < 0.05), suggesting favourable immunomodulatory action of vitamin C therapy in HAM patients. PMID- 8229034 TI - Low dose oral methotrexate treatment of multiple sclerosis: a pilot study. AB - An 18-month double-blind treatment of multiple sclerosis with low dose oral methotrexate showed it to be well tolerated and suggested effectiveness in exacerbating-remitting MS but not in the exacerbating progressive and chronic progressive stages. PMID- 8229035 TI - Methylprednisolone in multiple sclerosis: a comparison of oral with intravenous therapy at equivalent high dose. AB - A randomised double-blind placebo-controlled trial of intravenous methylprednisolone versus oral methylprednisolone at equivalent high dose was carried out on 35 patients with an acute relapse of multiple sclerosis (MS). After baseline evaluation each was randomly allocated to oral treatment and intravenous placebo or intravenous treatment and oral placebo, receiving 500 mg of methylprednisolone for five consecutive days and with reassessment at days five and twenty-eight. There was no significant difference in response when disability or functional scores were compared in the two groups. Adverse effects were minor and equally distributed. In this study oral treatment with methylprednisolone was as effective as intravenous treatment in acute relapse of MS. PMID- 8229036 TI - Dystonia in central pontine myelinolysis without evidence of extrapontine myelinolysis. AB - A 44-year-old female is described who developed persistent upper extremity and orolingual dystonia several weeks after suspected onset of central pontine myelinolysis (CPM), later confirmed by characteristic pontine lesions on MRI. No foci of the extrapontine myelinolysis were evident. This case confirms that dystonia may be a late and persistent sequela of CPM and may occur in the absence of visible lesions outside the brainstem. PMID- 8229037 TI - Thalamic experiential hallucinosis. AB - Two patients with an infarct limited to the thalamus developed auditory and visual experiential hallucinations. Neuropathological studies in one patient showed a small cavity in the right intralaminar nuclei surrounded by focal spongiform change, partly involving the right dorsomedial nucleus. Neuroradiological data in another patient indicated that the same nuclei in the left thalamus were also affected. It was concluded that a unilateral thalamic lesion could cause experiential hallucinations and the intralaminar and dorsomedial nuclei might be important structures to explain the phenomenon. PMID- 8229038 TI - Neuropsychological and SPECT scan findings during and after transient global amnesia: evidence for the differential impairment of remote episodic memory. AB - A patient had neuropsychological testing during, and at two days and seven weeks after a transient global amnesia (TGA) attack. During the attack she exhibited a characteristically profound anterograde amnesia but a limited remote memory loss; the most striking impairment was a deficit in personal episodic memory revealed by her performance on the Autobiographical Memory Interview. Personal and general semantic information was less impaired although there were indications of a temporal gradient in the impairment. When tested after the attack, she demonstrated normal anterograde and retrograde memory. A SPECT scan performed during TGA showed a focal reduction in cerebral perfusion in the postero-medial temporal lobes bilaterally which had resolved after seven weeks. PMID- 8229039 TI - Low incidence of seizures in patients with chronic subdural haematoma. AB - A total of 129 patients treated for chronic subdural haematoma were studied retrospectively to evaluate the incidence of seizures. None of 73 patients who were given prophylactic antiepileptic drug treatment developed seizures. Only two of 56 patients not given prophylaxis, developed early postoperative seizures. In these two, surgical technique was thought to be responsible. One patient developed complex partial seizures preoperatively. The incidence of seizures was therefore low, and similar to that previously reported for minor head injury. This study suggests that routine use of antiepileptic prophylaxis is not justified in patients with chronic subdural haematoma caused by minor head injuries, or other causes when there are no additional lesions present on CT scans. PMID- 8229040 TI - Transient partial verbal amnesia. PMID- 8229041 TI - Systemic effect of local botulinum toxin injections unmasks subclinical Lambert Eaton myasthenic syndrome. PMID- 8229042 TI - Electrophysiological improvement after intravenous immunoglobulin in motor neuropathy with multifocal conduction block. PMID- 8229043 TI - Glial cell abnormalities in the CNS of the carbonic anhydrase II deficient mutant mouse. AB - The Car-2n/Car-2n mutant mouse is entirely lacking in carbonic anhydrase II (CA), which is normally found in myelin and glial cells in the CNS. CNS tissue from CA deficient mutant mice was examined to see whether abnormalities could be detected which might help in understanding the function(s) of CA in the normal brain. Analyses of myelin yields and myelin proteins showed no differences between mutants and normal littermates. To visualize the oligodendrocytes and astrocytes in tissue sections, immunocytochemical staining was performed using antibodies against the Pi form of glutathione-S-transferase and glial-fibrillary acidic protein, respectively. In both gray and white matter from the mutants' brains the oligodendrocytes appeared to be shrinking and, possibly, degenerating. Hypertrophy of astrocytes occurred in the white matter, and the astrocytes in gray matter appeared swollen. It is suggested that imbalances in the HCO3-/CO2 ratios in various glial-cell compartments may produce abnormal distributions of water and ions in the brains of the CAII-deficient mice. PMID- 8229044 TI - Posthypoxic amnesia: regional cerebral glucose consumption measured by positron emission tomography. AB - Using positron emission tomography (PET), the regional cerebral metabolic rate of glucose consumption (rCMRGlc) was measured in 7 subjects suffering from chronic posthypoxic amnesia and in 12 controls. In 6 of the 7 patients regional decreases in rCMRGlc below normal limits were found involving the thalamus in 3 cases, the medial temporal cortex in 2 cases, and the caudate nucleus and cerebellum in one case each. No significant decrease in rCMRGlc was observed in any of the neocortical regions in any of the patients studied. Significant differences between group means of rCMRGlc were found only for the thalamus and the medial temporal cortex. These data suggest that posthypoxic amnesia stems from damage to the medial temporal cortex and its thalamic projection areas and not from neocortical dysfunction, confirming recent theories on the genesis of amnesia in man. PMID- 8229045 TI - Disorders of neuromuscular transmission due to natural environmental toxins. PMID- 8229046 TI - Effect of spatial frequency on transient and steady-state VEPs: stimulation with checkerboard, square-wave grating and sinusoidal grating patterns. AB - We recorded VEPs to the alteration of checkerboard, square-wave grating and sinusoidal grating patterns to evaluate the contribution of the fundamental spatial frequency and higher harmonic components in 12 normal subjects. Their fundamental spatial frequencies were equated and ranged from 0.5 to 8.0 c/deg. Both the transient VEP (T-VEP) and steady-state VEP (S-VEP) were obtained. The latency and amplitude of P100 of T-VEPs were measured. S-VEPs were Fourier analyzed, and phase and amplitude of the second harmonic response were measured. The mean P100 latency and the mean phase had a U-shaped function with a peak at a fundamental spatial frequency of 2.0 c/deg irrespective of the stimulus patterns, while the mean P100 amplitude and the mean amplitude of S-VEPs did not show such spatial selectivity. At low and medium spatial frequencies, differences in P100 latency, phase and amplitude between pattern types were more pronounced. However, this difference became insignificant at high spatial frequencies. These results indicate that VEP responses are predominantly determined by the fundamental spatial frequency, and that the contribution of higher harmonics to VEP responses is not negligible. Our results are consistent with the concept that the human visual system may use spatial frequency-domain information. PMID- 8229047 TI - Serum levels of ascorbic acid (vitamin C) in patients with Parkinson's disease. AB - To elucidate the possible role of vitamin C in the risk for developing Parkinson's disease (PD), we compared serum levels of ascorbic acid (vitamin C), measured by a fluorometric method, of 63 PD patients using their spouses as the control group. The serum levels of vitamin C did not differ significantly between the groups (47.13 +/- 0.89 micrograms/ml for PD and 47.60 +/- 0.60 micrograms/ml for controls). There was no influence of antiparkinsonian therapy on vitamin C. Serum levels of vitamin C did not correlate with age, age at onset and duration of the disease, scores of the Unified PD Rating Scale or the Hoehn and Yahr staging in the PD group. These results suggest that serum vitamin C concentrations are apparently unrelated to the risk of developing PD. PMID- 8229048 TI - Cerebrospinal fluid 28-kDa calbindin-D as a possible marker for Purkinje cell damage. AB - To examine the clinical value of 28-kDa calbindin-D (CaBP) in cerebrospinal fluid (CSF) as a marker for the damage to Purkinje cells, we measured CSF CaBP levels using an enzyme immunoassay method in 107 patients with cerebellar and other neurological diseases, and 26 controls. The mean CaBP level was markedly elevated in patients with cerebellar diseases, and the elevation of CaBP level was more frequent in the diseases involving Purkinje cells, such as multiple system atrophy (MSA) and subacute cerebellar degeneration in association with lung cancer. Further, in MSA patients, the CaBP levels decreased with duration of illness. The mean levels of CaBP were also elevated in some of the other diseases. We conclude that the elevations of CaBP levels are not specific for cerebellar diseases, but CSF CaBP may be a useful marker for examining the Purkinje cell involvement in cerebellar diseases. PMID- 8229049 TI - Mercury and selenium contents in amyotrophic lateral sclerosis in Hokkaido, the northernmost island of Japan. AB - We evaluated the pathogenicity of mercury (Hg) and selenium (Se) which are supposed to be one of the risk factors in the development of amyotrophic lateral sclerosis (ALS). Hg and Se contents were measured in plasma, blood cells, scalp hair samples of 21 sporadic ALS patients and 36 controls, who included 19 patients with other neurological diseases, in Hokkaido, the northernmost island of Japan. Hg and Se levels in plasma and blood cells of ALS patients were significantly lower in advanced staged ALS patients than controls. Low Hg and Se contents in ALS, being correlated with their disabilities and nutritional conditions, would rather reflect the disease contracted states than the pathogenic roles in ALS. PMID- 8229050 TI - Onset, natural history and outcome in idiopathic adult motor neuron disease. AB - Cases of adult-onset idiopathic motor neuron disease (MND) identified from January 1970 through December 1986 were studied in a defined area of California. The patients were followed prospectively throughout the illness in 99% of cases. Among 708 cases aged 25-74 years at onset, the most common type (86%) was typical, sporadic amyotrophic lateral sclerosis (SporALS). The risk of bulbar onset and shorter survival times increased with age in both men and women. About 4%, mainly younger men, experienced unusually long courses with milder paralysis, but could not be identified early in the illness. They probably represent one extreme of the ALS spectrum rather than a distinct subtype. Familial ALS (FamALS) was diagnosed in 7%. It developed earlier in life but ran a slightly longer course, which suggests a different disease process. Overall there was a statistically significant predominance of males, especially in 17 cases (2%) of progressive muscular atrophy (PMA). There were 26 cases (4%) classified as primary lateral sclerosis (PLS). Progressive bulbar palsy was not found; that diagnosis usually denotes merely the bulbar onset of ALS. PMID- 8229051 TI - Immunohistological evidence for second or somatic mutations as the underlying cause of dystrophin expression by isolated fibres in Xp21 muscular dystrophy of Duchenne-type severity. AB - Using five monoclonal antibodies against different parts of the dystrophin molecule, we have studied the dystrophin composition of 17 dystrophin-positive fibres in a muscle biopsy from a boy with Xp21 muscular dystrophy of Duchenne type severity. The fibres showed five distinct, reproducible, immunoreactive dystrophin profiles. All the profiles included both the N-terminal and the C terminal domains, but between these domains, different fibres were negative for different antibodies, suggesting the somatic loss of certain exons. We interpret this as the first in situ evidence of an individual having different patterns of missing exons leading to restoration of the reading frame in various ways in the original germline frame-shifting deletion of exons 35-43. It follows that various somatic mutations had taken place in different fibres. PMID- 8229052 TI - Intrathecal synthesis of specific antibodies in neuroborreliosis. Comparison of different ELISA techniques and calculation methods. AB - The sensitivity of six different ELISA techniques and calculation methods for the determination of intrathecal synthesis of IgG antibodies specific to Borrelia burgdorferi was investigated in paired CSF and serum specimens from 33 patients with neuroborreliosis. The diagnostic value of the Antibody Index (AI) was compared with the meaningfulness of serum antibodies to B. burgdorferi (Bb), established by immunofluorescence assay (IFA). The AI, as a standard for intrathecal antibody synthesis was determined from specific antibody ratios (QBb) in the CSF and serum and the CSF/serum ratio of IgG (QIgG) or albumin (QAlb). Using Western blotting with identical concentrations of IgG in the CSF and serum all patients displayed intrathecal synthesis of specific antibodies to at least two B. burgdorferi proteins. The different ELISA methods and calculation procedures were almost equivalent in demonstrating intrathecal synthesis of specific antibodies (32 and 33/33). Calculation of AI from IFA titers was somewhat less sensitive (30/33). In 5 patients titers of serum IgG- and IgM antibodies to B. burgdorferi determined by IFA were within the normal range or borderline, while elevated AIBb values indicated an autochthonous immune response to B. burgdorferi in the CSF. In uncertain cases of neuroborreliosis calculation of AI from ELISA titers will be useful in clarifying the diagnosis. PMID- 8229053 TI - Decreased IL-3 production by peripheral blood mononuclear cells in patients with multiple sclerosis. AB - The production of interleukin-3 by peripheral blood mononuclear cells (MNC) was assessed in patients with relapsing multiple sclerosis (MS) in both the active and the stable state, and in healthy controls. IL-3 levels were compared to levels of production of interleukin-2 (IL-2), tumor necrosis factor (TNF) and gamma-interferon (gamma-IFN). No significant differences in IL-3 levels were observed between stable-state patients and controls. When levels of cytokine production of patients in the inactive phase were compared to those of the same patients during relapse a significant decrease in IL-3 levels was observed, as opposed to significant increases in gamma-IFN and TNF levels, and an increase, though a non-significant, in IL-2 levels. The functional significance of lowered IL-3 production is unknown. However, the findings support the hypothesis of a highly complex interaction of overlapping regulatory influences within the cytokine network which parallels MS disease activity. PMID- 8229054 TI - An immunologic abnormality common to Bickerstaff's brain stem encephalitis and Fisher's syndrome. AB - The nosological position of Bickerstaff's brain stem encephalitis (BBE) has yet to be established, and its etiology is not clear. Because anti-GQ1b antibody frequently occurs in patients with Fisher's syndrome (FS) and there are clinical similarities between FS and BBE, we investigated anti-ganglioside antibodies in sera from 3 BBE patients who had transient long tract signs in addition to acute ophthalmoplegia and cerebellar-like ataxia in order to clarify the etiology and nosological position of BBE. High IgG anti-GQ1b antibody titers were present in all 3 sera samples but decreased with the clinical course of the illness. In contrast, no anti-GQ1b antibody was found in sera from patients with other neurologic diseases which were able to produce transient brain stem disturbance: multiple sclerosis, neuro-Behcet's disease, brain stem infarction, herpes simplex virus encephalitis, and Wernicke's encephalopathy. The finding that BBE and FS shared common autoantibody suggests that autoimmune mechanism common to FS is likely in BBE, and that both conditions represent a distinct disease with a wide spectrum of symptoms that include ophthalmoplegia and ataxia. PMID- 8229055 TI - Sympathetic skin response in multiple sclerosis. AB - Somatosympathetic reflex was studied in 29 patients with definite multiple sclerosis (MS) by the non-invasive sympathetic skin response (SSR) method. Abnormal SSRs in 1 or more limbs were noted in 17 patients. Good correlation between the number of absent SSRs and the severity of the disability caused by MS was observed. Delayed or absent SSRs correlated with leg weakness, spinothalamic sensory deficits, and neurogenic bladder. No correlation was noted between signal abnormalities seen on magnetic resonance imaging studies of the hypothalamus or brain stem and absent or delayed SSRs. It is suggested that damage to the central sympathetic fibers in the spinal cord accounts for the absent SSR. The damaged sympathetic fibers are probably located in the lateral columns of the spinal cord. PMID- 8229056 TI - A new neurocutaneous syndrome possibly related to Ota's nevus. AB - The case of a 42-year-old Sudanese man is reported who had developed hyperpigmentation of the left face and rapidly progressive right hemiparesis followed by myoclonus at the age of 30. Blue-grey hyperpigmentation was noted in the territory of the left ophthalmic, maxillary and mandibular trigeminal nerve branches, with alopecia of the right face and axilla. CT of the brain revealed left-sided paraventricular calcifications and MRI showed atrophy of the left cerebral hemisphere and mesencephalon, a calcified haemangioma in the left basal ganglia, an old infarct in the right central pons, and cerebellar atrophy. This case resembles Ota's syndrome; however, atypical distribution of the facial nevus, the epidermal location of hyperpigmentation, and the combination of vascular and degenerative cerebral malformations indicate that this is a new hitherto unreported neurocutaneous vascular syndrome. PMID- 8229057 TI - Effect of torbafylline on mitochondrial calmitine in mouse skeletal muscle regeneration after injection of a myotoxic drug. AB - The effect of torbafylline, a xanthine derivative (Hoechst, Werk Albert, Wiesbaden, Germany), was tested relative to changes in degeneration and subsequent regeneration processes in mouse gastrocnemius muscle induced by a single injection of chlorpromazine, a myotoxic drug. These processes were monitored by measuring changes in calmitine, a mitochondrial protein. We determined in our previous work that calmitine concentration decreases during degeneration and progressively increases during regeneration. In this study, we compared effects in torbafylline-treated mice with those in control mice treated with saline solution. The results show that regeneration is much faster with torbafylline treatment. Calmitine is decreased and returns quickly to normal in torbafylline-treated mice as compared to those treated with saline solution. Torbafylline might thus prove effective in stimulating muscle regeneration in myopathy. PMID- 8229058 TI - Sequential neuronal and astrocytic changes after transient middle cerebral artery occlusion in the rat. AB - The temporal evolution and spatial distribution of ischemic cell injury was investigated after transient middle cerebral artery (MCA) occlusion. Male Wistar rats (n = 61) were subjected to 2 h of MCA occlusion induced by advancing a nylon monofilament into the right internal carotid artery. Animals were killed after different durations of reperfusion, ranging from 4 to 166 h (n = 6-11 for each group). Neuronal injury and astrocytic reaction were evaluated using hematoxylin and eosin (H & E) and glial fibrillary acidic protein (GFAP) immunohistochemistry, respectively. Eosinophilic neurons were detected at 4 h of reperfusion in the basal ganglia, and at 10 h of reperfusion in the cortex. Focal brain infarct developed by 46 h of reperfusion, both in the cortex and the basal ganglia, and the volume remained constant between 46 and 166 h of reperfusion. Significant differences in astrocytic reaction were detected between the lesion and the periphery of the lesion at reperfusion times from 46 to 166 h; GFAP staining decreased in the core of the lesion and increased in the peripheral areas. Our data suggest that, after 2 h of MCA occlusion, brain tissue progresses from isolated neuronal injury to infarct with a time course dependent on anatomical site; and astrocytic reactivity, expressed by GFAP staining, reflects the outcome of the ischemic injury. PMID- 8229059 TI - Origin of macrophages in central nervous tissue. A study using intraperitoneal transplants contained in Millipore diffusion chambers. AB - In order to determine the origin of brain phagocytes brain slices and optic nerve segments from adult Lewis rats were transplanted into the peritoneal cavity of syngenic recipients. The specimens were contained in Millipore diffusion chambers fitted with membranes of either 0.22 or 5.0 microns pore size. The either blocked or allowed the access of non-resident cells. Each recipient rat received both a 0.22 and 5.0 microns pore chamber. Later (3-16 days), the specimens were recovered and analyzed by monoclonal antibody techniques and electron-microscopy. Endothelia, GFAP+ astrocytes, ED1-/ED2+/RCA-1+/OX-6-perivascular cells and ED1 /ED2-/RCA-1+/lysozyme--microglia were found to have survived the procedure. Cells of the macrophage phenotype (ED1+/ED2+/RCA-1+/lysozyme+/vimentin+ with phagocytic vacuoles), however, were only found in large numbers in specimens kept within 5.0 microns pore size chambers, giving access to non-resident cells, and were exceedingly rare in specimens from 0.22-micron pore chambers. It has been concluded that the majority of brain phagocytes found after lesions do not originate from microglia or perivascular monocytic cells, but rather from invading cells. PMID- 8229060 TI - A multiparametric data analysis showing the potential of localized proton MR spectroscopy of the brain in the metabolic characterization of neurological diseases. AB - We conducted an extended clinical evaluation of localized proton magnetic resonance spectroscopy (MRS) of the brain, performed on various brain diseases using short stimulated echo times. Pathologies studied were mainly multiple sclerosis, stroke, leukoaraiosis, AIDS-related leukoencephalopathies and glial tumors. Other miscellaneous pathologies were also studied. Magnetic resonance examination of the brain was conducted on a Siemens Magnetom SP63 (equipped with a 1.5 T magnet). Localized proton MRS was performed on a routine basis immediately after imaging, using the STEAM (stimulated echo acquisition mode) with a short echo time (20 ms) combined with a CHESS (chemical shift selective excitation) sequence. One or two VOI (8 ml) were examined. Data on 125 spectra were processed by principal component analysis (PCA) and conventional variance analysis. The following metabolite resonances were studied: inositol-glycine, taurine-scyllo-inositol, choline derivatives, phosphocreatine-creatine, aspartate, glutamine glutamate, N-acetylaspartate, acetate and lactate. PCA demonstrates that the different metabolic variables are independent. The analysis of groups of spectra clearly demonstrates that the metabolic profiles detected by localized MRS in various pathologies (i) differ significantly from controls, and (ii) allow a metabolic discrimination between groups of pathologies. Results of PCA are confirmed by variance analysis. Strokes are characterized by an increase in lactate concentration and leukoaraiosis by a decrease in inositol-glycine resonance. AIDS-related leukodystrophies are characterized by increases in lactate and choline concentrations. Reduction in N-acetylaspartate which is observed in most pathologies is not significant in the small lesions of white matter. Lactate has often been found in MS plaques, but no variation in the choline/phosphocreatine ratio was observed. GABA was tentatively assigned in the spectrum of a patient with epilepsy under sodium valproate treatment. This study illustrates the clinical feasibility of the technique, the value of a multiparametric data analysis in the definition of the pertinent variables characterizing the metabolic impairment, and the impact of localized proton MR spectroscopy of the brain in the assessment of cerebral suffering. PMID- 8229061 TI - Transcranial magnetic stimulation mapping of the motor cortex in normal subjects. The representation of two intrinsic hand muscles. AB - The TMS-mapped representations of two intrinsic hand muscles, abductor pollicis brevis (APB) and abductor digiti minimi (ADM), were quantified using a transcranial magnetic stimulation (TMS) mapping technique in 10 right handed and 6 left handed subjects. A 50 mm diameter figure eight coil was used. Stimulus sites were located using a latitude/longitude based coordinate system, stimulus intensity was threshold-adjusted and stimuli were applied during controlled low level (10%) voluntary contraction of the target muscles. Maps of the corticomotor representation were generated by fitting a continuously defined three dimensional function to the data obtained from stimulation at specific scalp sites, and projecting this function onto a two dimensional surface using a radial projection. It was found that the mapped representations of APB and ADM were large and overlapping but that there was a statistically significant separation of the two areas, the APB area being more laterally placed than the ADM area. The TMS-mapped representations of the two muscles showed no significant interhemispheric differences and were similar in left and right handed subjects. Rotation of the magnetic coil through 90 degrees resulted in medial shift and elongation of the TMS-mapped representations but there was no change in the relative positions of the two areas. The TMS-mapped representations were found to be very reproducible when mapping was repeated after intervals of up to 181 days. The present technique of TMS mapping allows the representation of individual hand muscles in the primary motor cortex to be reliably and reproducibly mapped and should prove useful for further studies of the physiology and pathophysiology of the motor cortex in man. PMID- 8229062 TI - Three cases of juvenile onset HTLV-I-associated myelopathy with pseudohypoparathyroidism. AB - Among 201 patients diagnosed with HAM/TSP at Kagoshima University, 21 juvenile onset patients had manifested clinical signs and symptoms at less than 15 years of age. They appeared to have common characteristics such as short stature and slight mental retardation. These signs prompted us to investigate five of them endocrinologically; and three patients with pseudohypoparathyroidism (PHP) were confirmed. Serum calcium levels were low, and human parathyroid hormone (PTH) infusion (Ellsworth-Howard test) caused low response in urinary cyclic AMP and phosphorus excretion. The first case had IgA nephropathy, which is generally associated with infectious diseases, while the second case had muscular lymphocytic infiltration. The mothers of cases 1 and 2, who were both seropositive for HTLV-I, were suspected to have abnormal calcium metabolism based on Ellsworth-Howard test. A brother of case 1 and two sisters of case 3 had also HAM/TSP and short stature. The early clinical onset of HAM/TSP may be due to PTH receptor anomaly and a low level of 1,25-dihydroxyvitamin D, which is deficient in PHP and is involved in the regulation of the immune response. The association with IgA nephropathy or myositis may result from progressive HTLV-I infection. PMID- 8229063 TI - Stimulation frequency-dependent neuromuscular junction transmission defects in patients with prior poliomyelitis. AB - Generalized fatigue and muscle fatiguability are major symptoms of post poliomyelitis syndrome (PPS), and may be due to neuromuscular junction transmission defects, as suggested by increased jitter on single fiber electromyography (SFEMG). To determine the etiology of this defect, we studied jitter at low (1, 5 Hz) and high (10, 15, 20 Hz) frequency stimulation with stimulation SFEMG in 17 post-polio patients with muscle fatiguability, and in 9 normal controls. In 5 of 17 PPS patients and in 1 of 9 controls, jitter was significantly higher (unpaired t-test, P < 0.05) at high frequency stimulation (HFS). In the remaining PPS patients and controls there was no significant difference in jitter at high and low stimulation frequencies. PPS patients with increased jitter at HFS had a significantly longer time interval since acute polio (mean 48.5 years) than PPS patients without increased jitter at HFS (mean 40 years; P < 0.05), but were not distinguished by other historical or clinical criteria. We conclude that the neuromuscular junction defect in post-polio patients is similar to that observed in amyotrophic lateral sclerosis, and is probably due to ineffective conduction along immature nerve sprouts and exhaustion of acetylcholine stores. The appearance of an increase in jitter with HFS in post-polio patients may be dependent upon time after acute polio. PMID- 8229064 TI - Human peripheral nerve macrophages in normal and pathological conditions. AB - We investigated, by immunocytochemistry and immune electron microscopy, the immunophenotype, morphology and functional properties of human peripheral nervous system (PNS) macrophages (M phi) under normal and pathological conditions. Endoneurial M phi disclosed an elongated, ramified morphology, with the main processes oriented along the major axis of nerve fibers; they shared several lineage-related and functional markers with monocyte/macrophages and central nervous system (CNS) microglia, including CD4, CR3, CR4 and FcRIII. In addition, basal expression of HLA-DR antigens was exclusively confined to M phi in normal PNS. In the course of unrelated pathological conditions, resident M phi underwent activation with transformation to hypertrophic cells or foamy phagocytes and up regulation of the markers expressed in normal conditions; new expression of a macrophagic antigen was detected on activated M phi. In different neuropathies, HLA-DR expression was also detected on non-myelin forming Schwann cells with ultrastructural features indicative of denervation. The present results demonstrate that the human PNS is provided with an intrinsic population of immunocompetent and potentially phagocytic M phi, which represent the peripheral counterpart of CNS microglia. PMID- 8229065 TI - Evidence for autoimmunity in amyotrophic lateral sclerosis. AB - Although the etiology and pathogenesis of ALS is unknown, increasing evidence supports a role for autoimmune mechanisms in motoneuron degeneration and death. An animal model, experimental autoimmune gray matter disease, can be induced by the inoculation of spinal cord gray matter. The experimental disease is characterized by weakness secondary to the loss of upper and lower motoneurons, accompanied by inflammatory foci within the spinal cord, and IgG at the neuromuscular junction and within UMN and LMN. In human ALS, IgG is present within the UMN and LMN, and T-lymphocytes and activated microglia have been identified within spinal cord gray matter and motor cortex. ALS IgG can passively transfer physiological changes of the neuromuscular junction to mice resulting in enhanced release of acetylcholine. The ALS IgG selectively interact with calcium channels and alter channel function. These data suggest a potential role for autoimmune mechanisms in the destruction and loss of motoneurons in ALS. PMID- 8229066 TI - Proto-oncogene c-fos induction in thiamine-deficient encephalopathy. Protective effects of nicardipine on pyrithiamine-induced lesions. AB - Treatment of rats with the central thiamine antagonist, pyrithiamine, results in severe neurological symptoms such as ataxia and convulsions. Induction of proto oncogene c-fos expression, often related to seizure activity, has been detected in the brains of thiamine-deficient rats by means of Northern blot analysis and in situ hybridization. Region-selective increases of lactate observed following thiamine deficiency development are largely coincident with histologically vulnerable regions. When thiamine-deficient rats were treated with the calcium channel blocker, nicardipine, lesions associated with thiamine deficiency did not appear and there was no induction of c-fos mRNA expression. This suggests a neurocytoprotective role of nicardipine to neuronal cell damage in thiamine deficient encephalopathy. PMID- 8229067 TI - Treatment of complex I deficiency with riboflavin. AB - We have evaluated the effects of treatment with riboflavin in five patients with a mitochondrial myopathy, associated with a complex I (NADH dehydrogenase) deficiency. Two patients suffered from a clinically pure myopathy and the other patients presented with encephalomyopathic features. Treatment with riboflavin resulted in a clear clinical improvement in the two patients with the myopathic form of complex I deficiency. However, only one of the patients with the encephalomyopathic form improved during therapy. In three of the four patients in whom complex I activity in muscle tissue has been determined again during therapy, complex I activity appeared to be normalized. The clinical effects of treatment in this group of patients do not correlate well with normalization of complex I activity. PMID- 8229068 TI - Blood-brain barrier permeability and the brain extracellular space in acute cerebral inflammation. AB - The diffusion properties of the brain cortical extracellular space have never been examined in models of inflammation, even though inflammation can cause increased blood-brain barrier permeability. Uptake of intravascular 125I-labelled albumin and the diffusion of the tetramethylammonium ion within the brain extracellular space was measured in an experimental brain abscess to determine the effect of acute inflammation upon blood-brain barrier permeability and diffusion properties of the cortical extracellular space. The blood-brain transfer constant for albumin was increased in the abscess region, indicating that an increase in blood-brain barrier permeability occurred in animals inoculated with a weakly pathogenic strain of Staphylococcus aureus. The volume fraction of the extracellular space, as measured by the diffusion of tetramethylammonium ion, ranged from 0.19 to 0.23 in bacteria inoculated subjects and from 0.21 to 0.22 in controls. The tortuosity of the extracellular space ranged from 1.40 to 1.42 in bacteria inoculated subjects and was 1.39 in controls. These results showed that the volume fraction and tortuosity of the cortical extracellular space were not affected by inflammation even though vascular permeability was increased. This result was supported by the finding that brain water content, measured in the same animals, was increased to a non significant extent in the bacteria inoculated subjects. These findings lead to the conclusion that acute inflammation induced by a weak pathogen can cause increased blood-brain barrier permeability without a significant change in the diffusion properties of the brain cortical space. PMID- 8229069 TI - Serotonergic regulation of the spinal cord content of thyrotropin releasing hormone in the cerebellar ataxia mutant mouse. AB - The effects of serotonin (5-HT) and various serotonin receptor antagonists on the spinal cord thyrotropin releasing hormone (TRH) content were studied in the rolling mouse Nagoya (RMN) and in the unaffected C3H mouse. TRH was extracted from the cervical, thoracic, lumbar, and sacral cord, at 1 h after the intraperitoneal injection of a serotonin precursor, 2 serotonin agonists, and 5 serotonin receptor antagonists. Administration of 5-hydroxytryptophan and 2 methyl-5-hydroxytryptamine resulted in an increase of the spinal cord TRH content in C3H mice, but not in RMN. Parachlorophenylalanine decreased the spinal cord TRH content in C3H mice, while it increased TRH levels in all regions of the RMN spinal cord. The TRH contents were decreased in all regions of the spinal cord after 5,7-dihydroxytryptamine administration in both C3H mice and RMN. In C3H mice, methysergide, mianserin, ketanserin, and spiperone significantly decreased the TRH content in all regions of the spinal cord, while 3 alpha-tropanyl-1H indole-3-carboxylic acid ester (ICS205-930) did not affect it. These antagonists paradoxically increased TRH levels in the cervical cord in RMN. The degradation of synthetic TRH by cord homogenates and the number and affinity of spinal cord serotonergic receptors (5-HT1 and 5-HT2) showed no significant difference between C3H mice and RMN. These results suggest that TRH turnover is abnormally regulated by serotonergic neurons in the RMN and that the dysfunction of the serotonergic nerves is attributable to the serotonin autoreceptor. PMID- 8229070 TI - Expression of the human groEL stress-protein homologue in the brain and spinal cord. AB - A monoclonal antibody (ML30), previously shown to identify a human mitochondrial protein epitope homologous with the groEL heat-shock protein of bacteria (hsp60), was used in an immunohistochemical survey of the central nervous system in patients dying with no evidence of neurological disease and in tissue from patients dying with various neurological disorders. Staining was performed on frozen tissue sections and on formalin fixed, paraffin embedded tissue. Astrocytes in all areas showed a strong pattern of punctate granular staining, which was increased in astrocytes showing reactive changes. Oligodendrocytes stained lightly in a diffuse granular pattern as did most neurons. Ependymal cells showed apical granular positivity. Expression of the hsp60 epitope recognised by ML30 was not seen in ubiquitinated inclusion bodies in motor neuron disease, neurofibrillary tangles in Alzheimer's disease or Lewy bodies in Parkinson's disease. The epitope recognised by ML30 was stable after formalin fixation and in post mortem tissue up to 96 h after death. Expression of the human groEL stress-protein homologue in brain and spinal cord is consistent with a mitochondrial location and may provide a morphological indicator of the functional or metabolic state of cells, especially glial cells. PMID- 8229071 TI - Myositis in Lyme borreliosis: an immunohistochemical study of seven patients. AB - Myositis is a rare complication of Lyme disease. In order to get information about the pathogenesis of this disorder, muscle specimens of 7 patients suffering from myositis as a manifestation of Lyme borreliosis were examined by immunohistology. Lyme spirochetes could be found in muscle biopsies of 6 patients. Infiltrates consisted mainly of macrophages and T helper/inducer cells. The T4/T8 ratio was 1.7 in the endomysium and 2.1 in the perimysium. Increased expression of MHC-I molecules by several muscle fibers was observed in 2 subjects only. No MHC-II molecules were expressed by muscle fibers. Lymphocytes expressing the interleukin-2 receptor were detected in 2 patients. Leu-15+ and Leu-11+ cells were found only to a slight extent in 2 patients. In conclusion the immunohistochemical findings in myositis due to Lyme borreliosis are different from other manifestations of this disease, and also from other forms of myositis. PMID- 8229072 TI - Detection of herpes simplex virus in the cerebrospinal fluid of patients with encephalitis using the polymerase chain reaction. AB - Herpes simplex encephalitis is a neurologic emergency demanding immediate institution of specific therapy in order to prevent mortality. Diagnosis, however, is a complex matter with controversy existing over the appropriateness of brain biopsy. We report the demonstration of herpes simplex virus DNA by means of the polymerase chain reaction (PRC) in the cerebrospinal fluid of 3 patients with herpes simplex encephalitis. One of the patients suffered from brain-stem encephalitis with high intensity signals in the pons on magnetic resonance imaging, the second reported case of this entity. The PCR for herpes simplex on a control series of cerebrospinal fluid of 20 patients with other central nervous system infections was negative. PCR of cerebrospinal fluid offers a sensitive, specific and rapid diagnosis of herpes simplex encephalitis, making brain biopsy unnecessary. Still, the importance of strict measures to prevent contamination cannot be stressed enough. It is possible that due to the high sensitivity of the PCR, herpes simplex may be found in other infectious syndromes of the central nervous system. PMID- 8229073 TI - Embolic infarcts of carotid origin differ in size and site with contralateral carotid patency. AB - Carotid emboli cause small infarcts in the ipsilateral hemisphere in rats when the contralateral carotid artery is fully patent. With contralateral carotid occlusion, embolic infarcts, both large and small, occur with equal proportions in both hemispheres. To determine if emboli also cross to the territory of a stenotic carotid artery, we combined high grade (78-96%) stenosis of the left common carotid artery with photochemically-induced (laser irradiation 632 nm, 200 mW/cm2, 15 min; intravenous Photofrin II, 12.5 mg/kg) embolism from the right common carotid artery in 12 rats. Ninety-eight cerebral infarcts occurred in 9 experimental animals, with eight infarcts being large (> 2.5 mm). The mean proportions of infarcts and emboli on the left were 25% and 19%, respectively. These results suggest that contralateral carotid artery stenosis, like occlusion, will influence the site and size of embolic infarcts and that the "symptomatic" carotid artery cannot always be determined by the side of the cerebral infarct. PMID- 8229074 TI - A review of neuronal damage in human immunodeficiency virus infection: its assessment, possible mechanism and relationship to dementia. AB - Over the past decade it has been realized that HIV affects the central nervous system, and various investigations have illuminated the spectrum of neuropathology in AIDS. One major advance has been the demonstration that there is substantial neuronal loss, which appears independent of the HIV-associated inflammatory lesions. Quantitative studies on neuronal populations, while fraught with methodological difficulties, are essential to the understanding of the mechanism of this neurotoxic damage. This article will review, firstly, the modern stereological procedures available for quantitative investigations; secondly, the pattern, degree and time scale of HIV-associated neuronal loss; thirdly, other morphological evidence of neuronal damage; and finally, the pathological and clinical implications of these findings. PMID- 8229075 TI - Brainstem serotonergic neurons in chronic alcoholics with and without the memory impairment of Korsakoff's psychosis. AB - There are several lines of evidence to suggest that serotonergic neurons in the brain are detrimentally affected by chronic alcohol consumption. The present study aims to quantify pathological changes in brainstem regions containing serotonergic neurons in chronic alcoholics compared to age-matched non-alcoholic controls. An antibody specific for tryptophan hydroxylase was used to immunohistochemically demonstrate serotonergic neurons in serial sections of postmortem brainstem. The cases analyzed were divided into four groups on the basis of their clinical and pathological presentation; chronic alcoholics with Wernicke's encephalopathy, chronic alcoholics with additional Korsakoff's psychosis, non-alcoholic controls, and a single chronic alcoholic without neurological complications. There was an overall reduction in the number of serotonergic neurons in all alcoholic cases when compared with controls. All brainstem regions were affected, but the largest neuronal loss was found in areas of the medullary and caudal pontine reticular formation (reduced by 80-90%). Alcoholics with Korsakoff's psychosis did not differ in the amount or extent of pathology from the other alcoholic cases analyzed. The data indicate that significant numbers of serotonergic neurons degenerate in chronic alcoholics. Such a loss is likely to have significant clinical consequences. PMID- 8229076 TI - Functional alterations in Alzheimer's disease: diminution of cytochrome oxidase in the hippocampal formation. AB - In Alzheimer's disease, the relationship between structural alterations such as neurofibrillary tangles and senile plaques and neuronal function is unknown. Cytochrome oxidase, the terminal enzyme of the electron transport, is a marker of neuronal functional activity. Its activity is diminished in experimentally deafferented neurons. Based on evidence that the molecular layer of the dentate gyrus is deafferented in the brains of individuals with Alzheimer's disease, we hypothesized that cytochrome oxidase activity would be diminished in this region secondary to reduced glutamatergic input. Using cytochrome oxidase histochemistry, we found a change in the distribution of cytochrome oxidase in the molecular layer of the dentate gyrus and a decrease in activity in both the dentate gyrus and hippocampal subfields in Alzheimer's disease. In contrast, we found relatively little structural pathology in the dentate gyrus, CA4, and CA3 in these individuals. These results suggest that neurons that remain structurally intact in Alzheimer's disease may nonetheless undergo changes in metabolic function as neural systems fail. PMID- 8229077 TI - Neuronal changes in the cerebral cortex of the rat following alcohol treatment and thiamin deficiency. AB - The contribution of thiamin deficiency to the pathology of alcohol-related brain damage is still unclear. This study used a model of prolonged alcohol abuse in which animals were subjected to a brief period of mild thiamin deficiency. The episode of thiamin deficiency was early (after 4 weeks), in the middle (after 15 weeks) or late (after 26 weeks) in their 28 week alcohol treatment period. A control group of animals fed no alcohol and maintained on a thiamin-replete diet was used for comparison. The brains were removed and sectioned in the coronal plane at 50 microns intervals. Successive serial sections were stained with cresyl violet for Niss1 substance and immunohistochemically with antibodies to the calcium-binding proteins parvalbumin and calbindin. These calcium-binding proteins identify the majority of GABA-containing neurons in the cerebral cortex. The number of cells in the Fr1 region of the cerebral cortex was quantitated. A significant loss of Niss1-stained neurons was identified from the early group, while a loss of parvalbumin-immunoreactive neurons was seen in the early and middle groups. No loss of neurons was identified from the late group. In addition, no loss of calbindin-immunoreactive neurons was seen. This study represents the first report of cortical neuronal loss in an animal model of alcohol abuse and thiamin deficiency. Moreover, the results imply that thiamin deficiency is integrally involved in the pathogenesis of alcohol-related cortical neuronal loss. PMID- 8229078 TI - The lack of accumulation of senile plaques or amyloid burden in Alzheimer's disease suggests a dynamic balance between amyloid deposition and resolution. AB - A beta, a nearly insoluble peptide, is generally assumed to irreversibly deposit and accumulate as senile plaques (SP) during the course of Alzheimer's disease (AD). We have studied temporal neocortex of normal elderly subjects, AD patients, and elderly Down syndrome (DS) patients to determine whether A beta accumulates with age or with increasing duration of illness. We measured the number, size distribution, and total area (amyloid burden) of A beta immunoreactive deposits using computerized image analysis techniques. We found far fewer SP in normal control subjects than in AD patients, who in turn have fewer SP than elderly DS patients. No measure of A beta correlated with age in the control subjects, nor duration of illness in AD or DS patients. These data indicate that A beta may not continue to accumulate during these disease processes and support the view that the amount of A beta observed at autopsy may reflect competing processes of deposition and resolution of amyloid. PMID- 8229079 TI - Morphometric analysis of microglia in Alzheimer's disease. AB - HLA-DR-immunoreactive microglia were quantitated in the middle temporal gyrus of five late-stage patients with Alzheimer's disease (AD) and five age-matched control subjects using LN3 immunocytochemistry and computerized morphometric image analysis. The grand mean numerical density of HLA-DR-immunoreactive microglia in AD (403.86/mm2) was significantly larger than in the five control subjects (193.33/mm2) (p < 0.0001). The grand mean cross-sectional area of HLA-DR immunoreactive microglia in AD subjects (222.90 microns2) was significantly larger than in control subjects (121.59 microns2) (p < 0.0001). The percentage of total microglia that were large activated microglia was much greater in AD (47.9%) than in control subjects (15.2%). The grand mean percent of the cortical area occupied by HLA-DR-immunoreactive microglia in AD (9.02%) was significantly greater than in controls (2.35%) (p < 0.0001). Microglia were present in greater numbers in the outer three cortical laminae in both AD and controls. The striking increase in activated microglia in the neocortex and their role in the inflammatory response and possible secretion of neurotoxins could be important in neuron degeneration in AD. PMID- 8229080 TI - Regional heterogeneity in the proliferative activity of human gliomas as measured by the Ki-67 labeling index. AB - The effects of regional heterogeneity on the accuracy of histological grading of gliomas are well known, but little has been reported about its implications for other diagnostic modalities. This study investigated the relationships of regional heterogeneity in tumor proliferative activity, measured by Ki-67 labeling indices (LI), and histological grades for 16 regionally sampled glioma resections. There was a strong correlation between histological grades and Ki-67 LI in individual regions (p < 0.001), and both methods demonstrated comparable heterogeneity. Heterogeneity increased with tumor grade, probably as an expression of the increased genetic instability that accompanies tumor progression. Similarly, regions with comparable proliferative activity tended to cluster, paralleling clonal expansion. Thus, both histological grading and Ki-67 LI are subject to heterogeneity-induced sampling errors that limit their diagnostic accuracy, particularly in small biopsies. However, fewer grading errors occurred when using both methods together than when using either method alone, suggesting that the use of multiple techniques may reduce the adverse effects of regional heterogeneity on diagnostic accuracy. Regional heterogeneity appears to be a ubiquitous feature of gliomas: it also has been reported in karyotype, p53 oncogene mutations, and PDGF and EGFR expression. The effects of regional heterogeneity on new methods for studying gliomas need to be considered. PMID- 8229081 TI - Re-evaluation of the structural organization of neuritic plaques in Alzheimer's disease. AB - We re-examined the relationship among synaptic pathology, subcellular abnormalities within neurites in the plaques and beta-amyloid deposits of Alzheimer's disease (AD) using laser confocal imaging and computer-aided serial section reconstruction techniques. Analysis of serial optical sections of neuritic plaques double-immunolabeled for anti-beta-amyloid/anti-tau-2 revealed that 35% of this type of plaque contained a dense amyloid core with clusters of peripheral abnormal neurites. The other 65% were without a dense core and were mainly composed of abundant abnormal neuritic clusters with bundles of amyloid distributed throughout the neuritic plaque. While two-dimensional (2-D) analysis of the plaques has suggested that neurites are distributed in the plaque periphery with beta-amyloid localized in its center, serial section analysis showed the opposite arrangement can also be true. Three-dimensional (3-D) reconstructions of serial optical sections showed that the tau-positive tortuous axons clustered in the neuritic plaques were often continuous with synaptophysin positive distended terminals. Analysis of electron micrographs from serial sections showed continuity among the different segments of the neurites. Further analysis of the computer generated 3-D reconstructed neuritic plaques (both from serial electron micrographs and serial optical sections), viewed as continuous rotating loops, confirmed that a great majority of the plaque volume was occupied by the clustered and continuous abnormal neurites, while the amyloid fibrils were compressed and displaced to the periphery of the plaque. The 3-D imaging of the neuritic plaques in AD suggests a more widespread and active neuritic damage than that predicted from 2-D observations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229082 TI - Disorder of cerebellar foliation in Walker's lissencephaly and neu-laxova syndrome. AB - A diffuse disorder of cerebellar foliation was found in eight infants and one fetus with Walker's lissencephaly. The cerebellar cortex consisted of fused and irregularly distorted folia. In the white matter, trilaminated rings of cortex were concentrically arranged around blood vessels and mesenchymal tissue. The normal relative position of the different classes of cortical nerve cells was preserved. Cells of the external granular layer invaded the meninges and migrated along penetrating blood vessels. We believe that this foliation disorder is caused by a defect in the external basal lamina that allows adjacent folia to be fused and sulci obliterated by intrameningeal ectopias of external granule layer cells. Physical forces applied during development probably contribute to the distortion of the gyral pattern. There was a volumetric reduction of the neocerebellum, which might also be a consequence of the basal lamina defect. The cerebellum of a fetus with the Neu-Laxova syndrome showed the same abnormalities as in Walker's lissencephaly. It is postulated that these two conditions belong to a class of prenatal developmental disorders that involves a defect of the extracellular matrix. PMID- 8229083 TI - beta-Amyloid protein is higher in Alzheimer's disease brains: description of a quantitative biochemical assay. AB - Deposition of beta-amyloid protein (A beta) in senile plaques and in the walls of cerebral vessels is a pathologic hallmark of Alzheimer's disease (AD). The current diagnostic criteria for AD requires the presence of neurofibrillary tangles and a minimum number of senile plaques in cortex. Senile plaques are readily visualized by silver staining or immunocytochemistry using antibodies raised to A beta. Available histochemical and immunocytochemical methods are sensitive but the results may occasionally be variable and sampling from many brain regions is difficult and impractical. This study describes a simple biochemical method for quantifying the A beta load in unfixed brain homogenates. The immunoassay recognizes all forms of A beta deposits (neuritic and diffuse plaques, and cerebrovascular amyloid) and has a sensitivity and specificity comparable to immunocytochemistry. In direct comparisons, results from the dot blot method correspond well with both Western blot analysis of partially purified A beta and plaque counting by immunocytochemistry. In a retrospective series of 39 postmortem AD and control cases, the amount of A beta in brain by dot blot immunoreactivity effectively separated the two groups. Therefore, this method provides a rapid, sensitive, and accurate quantitation of A beta in postmortem brain tissue and represents an alternative approach for studying A beta deposition in aging and AD. PMID- 8229084 TI - Plaque-only Alzheimer disease is usually the lewy body variant, and vice versa. AB - A minority of neuropathologically confirmed Alzheimer disease (AD) brains lack neocortical neurofibrillary tangles or have very few, constituting a form of "plaque-only AD." A significant percentage of clinically diagnosed AD patients are found at autopsy to have both AD and brainstem and neocortical Lewy bodies. Many of these Lewy body variants of AD (LBV) have numerous senile plaques but no neocortical neurofibrillary tangles, and so resemble plaque-only AD. In this study, we sought to determine if plaque-only AD was usually LBV, and, conversely, if LBV was usually plaque-only AD. We analyzed 147 consecutively accessioned cases of neuropathologically confirmed AD, diagnosed according to criteria from the National Institute on Aging and the Consortium to Establish a Registry for Alzheimer's Disease. Twenty-five percent of all AD cases in this series were plaque-only AD, and 75% were plaque and tangle AD. Twenty-eight percent of AD cases in this series were LBV, and 72% were pure AD. Of the plaque-only AD cases, 75% were LBV and only 25% were pure AD. Of the LBV, 66% were plaque-only AD and only 33% were plaque and tangle AD. These results indicate that most plaque-only AD is LBV, and, conversely, that most LBV is plaque-only AD. PMID- 8229085 TI - Differential localization of class III, beta-tubulin isotype and calbindin-D28k defines distinct neuronal types in the developing human cerebellar cortex. AB - This immunohistochemical study compares the localization of the neuronal class III beta-tubulin isotype (beta III) to that of calbindin-D28k in 40 human fetal and postnatal cerebella ranging from 12 weeks gestation to adulthood. In the external granule layer of the developing cerebellar cortex, beta III staining was present in the premigratory (postmitotic) zone of horizontal neurons but was absent in "epithelioid" cells of the subpial proliferative mitotic zone. In the molecular layer, intense beta III staining was associated with parallel fibers, stellate/basket neurons and migrating fusiform granule neurons. beta III staining was also present in internal granule neurons. In contrast, beta III was not detectable in fetal and neonatal Purkinje neurons and Golgi II neurons, but was evident in these neurons from juvenile and adult cerebella. Calbindin-D28k staining was present in Purkinje neurons also delineating their somatic spines ("pseudopodia"), lateralizing and apical dendrites (including dendritic spines), subpopulations of small to intermediate-sized Golgi II neurons in the internal granule layer ("synarmotic cells" of Landau), large to medium-sized subcortical Golgi II neurons and neurons of cerebellar roof nuclei, at various gestational stages and postnatally. It was absent in the external granule layer, parallel fibers, stellate/basket and internal granule neurons. Variable degrees of beta III and calbindin-D28k staining were detected in subpopulations of immature neuroepithelial cells of the ventricular matrix at the roof of the fourth ventricle. Glial (including Bergmann glia) and mesenchymal cells were not stained for either antigenic determinants. The differential expression of calbindin-D28k and beta III defines distinct populations of neurons in the developing human cerebellar cortex and supports the ontogenetic concept of Ramon y Cajal. PMID- 8229086 TI - In vitro studies of axonally-regulated Schwann cell genes during Wallerian degeneration. AB - Wallerian degeneration in vivo is associated with marked downregulation of myelin protein genes such as P(o) and upregulation of other genes such as nerve growth factor receptor (NGF-R), glial fibrillary acidic protein (GFAP) and neural cell adhesion molecule (N-CAM). This study examines the expression of these genes during Wallerian degeneration in vitro and how manipulating Ca2+ affects this response. Small explants of sciatic nerve from normal young adult rats cultured for five days show similar reversal of the myelinating phenotype as found in vivo. If Ca++ is removed from the culture medium through the addition of EGTA, expression of the nerve growth factor receptor and glial fibrillary acidic protein genes is inhibited but downregulation of the P(o) gene still occurs. Explants cultured in medium containing EGTA are still capable of expressing nerve growth factor receptor if the medium is replaced by one containing Ca2+. Supplementation of normal medium with drugs modulating Ca2+, such as Bepridil which blocks the Na+Ca2+ exchanger or compound 48/80 which inhibits calmodulin, also prevent the expression of the nerve growth factor receptor gene during Wallerian degeneration in vitro. Treatment of the cervical sympathetic trunk with Bepridil leads to loss of the nerve growth factor receptor immunoreactivity which is normally present. The results indicate that Ca2+ may play a role in the expression of the nerve growth factor receptor gene during Wallerian degeneration and provide some indication that this effect may be directly on the Schwann cell rather than operating indirectly via the axon. PMID- 8229087 TI - The effects of exercise training of different intensities on neuromuscular junction morphology. AB - Little is known about the effects of exercise training on neuromuscular junction morphology in skeletal muscle. The objectives of this investigation were: 1) to determine if exercise training would elicit changes in neuromuscular junction morphology, 2) to determine if exercise training of different intensities would evoke specific changes in neuromuscular junction morphology, and 3) to determine whether changes in neuromuscular junction structure occur independently of changes in muscle fibre type and size. Twenty-four age and size matched male Sprague-Dawley rats were randomly assigned to three groups: high-intensity trained (HIT), low-intensity trained (LIT), or untrained. Neuromuscular junction morphology of the soleus muscle was determined via immunofluorescent staining. Presynaptic acetylcholine vesicles were visualized with SV-2 antibody in conjunction with fluorescein isothiocyanate labelled secondary secondary antibody. Postsynaptic acetylcholine receptors were identified with rhodamine labelled alpha-bungarotoxin. Laser scanning microscopy was used to produce images of synapses, which were used to quantitate the following: total area of SV-2 and alpha-bungarotoxin staining, density of acetylcholine vesicles and receptors, structural complexity, and synaptic coupling. To visualize nerve terminal branching, a smaller number of neuromuscular junctions were stained with C-2 antibody, which reacts with a neurofilament epitope, in conjunction with fluorescein isothiocyanate labelled secondary antibody. Total length of branching, number of branches, average length of branches, and ratio of secondary to primary branches per neuromuscular junction were determined. Citrate synthase activity, fibre type composition and fibre cross-sectional areas of the soleus muscle were assessed to determine the presence of a training effect in that muscle. Results indicate that training did induce hypertrophy of the neuromuscular junction that was independent of muscle hypertrophy. Although the HIT and LIT groups exhibited similar hypertrophic responses of the neuromuscular junction, the HIT group displayed more dispersed synapses than the LIT group. Neither exercise training program, however, resulted in altered densities of acetylcholine vesicles or receptors, nor did training significantly change synaptic coupling. Nerve terminal branching was also affected by exercise training. Neuromuscular junctions from the HIT group demonstrated a greater total length of branching, average length per branch, and number of finer, or secondary, branches than those of the LIT group. PMID- 8229088 TI - The endoplasmic reticulum-associated VP7 of rotavirus is targeted to axons and dendrites in polarized neurons. AB - Rotavirus, which matures and is retained in the endoplasmic reticulum, was used to examine how polarized dorsal root ganglion and spinal cord neurons distributed cytoplasmic and endoplasmic reticulum-associated proteins. A remarkable observation was that NS28, a trans-endoplasmic reticulum-membrane protein which functions as a receptor for budding particles, remained in the cell body during the whole course of infection (48 h) while the VP7 glycoprotein, which is endoplasmic reticulum associated and usually retained in the endoplasmic reticulum, was targeted to axons already 4 h post infection. VP7 was furthermore transported in an endo-beta-N-acetylglucosaminidase H sensitive form through the secretory pathway. The segregated appearances of NS28 and the endo-beta-N acetylglucosaminidase H sensitive VP7 indicate that VP7 enters a transport compartment separate from NS28. Brefeldin A treatment rapidly disintegrated the Golgi apparatus of the neurons and rapidly blocked axonal transport of Sendai virus glycoproteins, while axonal transport of rotavirus VP7 was not blocked, suggesting that VP7 uses an intracellular pathway in neurons which does not involve the Golgi apparatus. PMID- 8229089 TI - Formation of synapses between basal forebrain afferents and cerebral cortex neurons: an electron microscopic study in organotypic slice cultures. AB - Co-cultures of rat basal forebrain and cerebral cortex were maintained from 1 to 5 weeks in vitro with serum-free defined medium. The formation of synaptic connections between basal forebrain afferent fibres and cortical neurons was studied by specific labelling with three staining techniques, including (i) neuronal tract tracing with the fluorescent dye 1,1'-dioctodecyl-3,3,3'3'- tetramethylindocarbocyanine perchlorate, (ii) acetylcholinesterase histochemistry, and (iii) choline acetyltransferase immunocytochemistry. Both basal forebrain and cerebral cortex tissue displayed organotypic characteristics in culture. Cerebral cortex revealed a dense innervation by axonal projections from the basal forebrain. All three labelling techniques produced similar results at the light microscopic level, with densest innervation located in the marginal zone. At the fine structural level, the 1,1'-dioctodecyl-3,3,3'3' tetramethylindocarbocyanine perchlorate-, acetylcholinesterase- and choline acetyltransferase-stained basal forebrain afferents all revealed a number of synaptic contacts with cortical neurons. The contacts displayed consistent synaptic features, including presynaptic accumulation of small round vesicles, cleft widening, and postsynaptic densities forming symmetric synapses. These morphological characteristics of connections formed in vitro are similar to basal forebrain cholinergic projections to cerebral cortex in normal brain. Based on these results, this tissue culture model appears to be an useful tool for investigations of the development of cholinergic innervation of cerebral cortex. PMID- 8229090 TI - Effects of excitatory amino acids on immunocytochemically identified populations of neurons in turtle retina. AB - Excitatory amino acids play an important role in visual processing in the retinas of many species, but little is known about the identity of the specific postsynaptic cell types and the pharmacology of their receptors. To investigate which specific cell types were affected by excitatory amino acids, we examined the effects of exogenous aspartate, glutamate, kainic acid, N-methyl-D-aspartate, and MK-801 on retinal neurons. Specific populations of neurons were labelled using antibodies directed against glucagon, enkephalin, neurotensin, gamma aminobutyric acid, glutamic acid decarboxylase, serotonin, glycine, glutamate or aspartate. We analyzed a combination of long-term in vivo injections (seven days following an intraocular injection of kainic acid) and short term in vitro incubations. There were changes in the labelling intensity and sometimes in the relative localization of all of the antigens in the drug treated retinas. Some observations suggested that the drugs were altering neurotransmitter metabolism. Differential responses were seen in specific cell types within the populations of neurons with neurotensin-, glutamate-, aspartate-, glycine, gamma-aminobutyric acid-, and glutamic acid decarboxylase-like immunoreactivity. The immunocytochemical approach used in these studies was able to determine specific retinal cell types which were influenced by particular excitatory amino acids. The broad extent of cell types influenced and the potential metabolic effects suggest that excitatory amino acids and their receptors play a complex role in visual processing. PMID- 8229091 TI - Close apposition among neighbouring axonal endings in a neuroma. AB - Axons in intact peripheral nerve trunks constitute independent afferent and efferent communication channels. However, when nerves are severed, several different forms of axon-axon cross-excitation develop in association with the injury site. In this study we have examined experimental sciatic nerve-end neuromas in rats with special interest in the compartmentalization of individual axons, and the barriers that separate close neighbours. At postinjury times at which functional coupling is known to occur, neuromas were found to contain many examples of axons in which adjacent membrane faces come into close contact without an intervening Schwann cell process. These occur in bundles containing from two to as many as 30 individual nerve fibres wrapped in a common Schwann cell sheath. The surface area of close apposition between axon pairs ranges up to several tens of micron2. Closely apposed axon profiles may be outgrowing branches of a single parent axon, but anterograde tracer data indicate that many belong to independent neurons. Closely apposed axons are separated from one another, and from associated Schwann cell processes, by a cleft about 130 A wide. No synapses, gap junctions or tight junctions were observed. Extracellular tracer studies using La3+ and Ruthenium Red indicated that the cleft system is patent, permitting the free diffusion of small molecules between the space adjacent to the axolemma and the bulk extracellular compartment. Together, these data provide a structural basis for interfibre interactions based on local electrical current flow (ephaptic crosstalk), as well as coupling mediated by K+ ions and neurotransmitter molecules. PMID- 8229093 TI - De novo and inverse folding predictions of protein structure and dynamics. AB - In the last two years, the use of simplified models has facilitated major progress in the globular protein folding problem, viz., the prediction of the three-dimensional (3D) structure of a globular protein from its amino acid sequence. A number of groups have addressed the inverse folding problem where one examines the compatibility of a given sequence with a given (and already determined) structure. A comparison of extant inverse protein-folding algorithms is presented, and methodologies for identifying sequences likely to adopt identical folding topologies, even when they lack sequence homology, are described. Extension to produce structural templates or fingerprints from idealized structures is discussed, and for eight-membered beta-barrel proteins, it is shown that idealized fingerprints constructed from simple topology diagrams can correctly identify sequences having the appropriate topology. Furthermore, this inverse folding algorithm is generalized to predict elements of supersecondary structure including beta-hairpins, helical hairpins and alpha/beta/alpha fragments. Then, we describe a very high coordination number lattice model that can predict the 3D structure of a number of globular proteins de novo; i.e. using just the amino acid sequence. Applications to sequences designed by DeGrado and co-workers [Biophys. J., 61 (1992) A265] predict folding intermediates, native states and relative stabilities in accord with experiment. The methodology has also been applied to the four-helix bundle designed by Richardson and co-workers [Science, 249 (1990) 884] and a redesigned monomeric version of a naturally occurring four-helix dimer, rop. Based on comparison to the rop dimer, the simulations predict conformations with rms values of 3-4 A from native. Furthermore, the de novo algorithms can assess the stability of the folds predicted from the inverse algorithm, while the inverse folding algorithms can assess the quality of the de novo models. Thus, the synergism of the de novo and inverse folding algorithm approaches provides a set of complementary tools that will facilitate further progress on the protein-folding problem. PMID- 8229092 TI - Prediction and analysis of structure, stability and unfolding of thermolysin-like proteases. AB - Bacillus neutral proteases (NPs) form a group of well-characterized homologous enzymes, that exhibit large differences in thermostability. The three-dimensional (3D) structures of several of these enzymes have been modelled on the basis of the crystal structures of the NPs of B. thermoproteolyticus (thermolysin) and B. cereus. Several new techniques have been developed to improve the model-building procedures. Also a 'model-building by mutagenesis' strategy was used, in which mutants were designed just to shed light on parts of the structures that were particularly hard to model. The NP models have been used for the prediction of site-directed mutations aimed at improving the thermostability of the enzymes. Predictions were made using several novel computational techniques, such as position-specific rotamer searching, packing quality analysis and property profile database searches. Many stabilizing mutations were predicted and produced: improvement of hydrogen bonding, exclusion of buried water molecules, capping helices, improvement of hydrophobic interactions and entropic stabilization have been applied successfully. At elevated temperatures NPs are irreversibly inactivated as a result of autolysis. It has been shown that this denaturation process is independent of the protease activity and concentration and that the inactivation follows first-order kinetics. From this it has been conjectured that local unfolding of (surface) loops, which renders the protein susceptible to autolysis, is the rate-limiting step. Despite the particular nature of the thermal denaturation process, normal rules for protein stability can be applied to NPs. However, rather than stabilizing the whole protein against global unfolding, only a small region has to be protected against local unfolding. In contrast to proteins in general, mutational effects in proteases are not additive and their magnitude is strongly dependent on the location of the mutation. Mutations that alter the stability of the NP by a large amount are located in a relatively weak region (or more precisely, they affect a local unfolding pathway with a relatively low free energy of activation). One weak region, that is supposedly important in the early steps of NP unfolding, has been determined in the NP of B. stearothermophilus. After eliminating this weakest link a drastic increase in thermostability was observed and the search for the second-weakest link, or the second-lowest energy local unfolding pathway is now in progress. Hopefully, this approach can be used to unravel the entire early phase of unfolding. PMID- 8229094 TI - Protein fold recognition. AB - An important, yet seemingly unattainable, goal in structural molecular biology is to be able to predict the native three-dimensional structure of a protein entirely from its amino acid sequence. Prediction methods based on rigorous energy calculations have not yet been successful, and best results have been obtained from homology modelling and statistical secondary structure prediction. Homology modelling is limited to cases where significant sequence similarity is shared between a protein of known structure and the unknown. Secondary structure prediction methods are not only unreliable, but also do not offer any obvious route to the full tertiary structure. Recently, methods have been developed whereby entire protein folds are recognized from sequence, even where little or no sequence similarity is shared between the proteins under consideration. In this paper we review the current methods, including our own, and in particular offer a historical background to their development. In addition, we also discuss the future of these methods and outline the developments under investigation in our laboratory. PMID- 8229095 TI - The importance of short structural motifs in protein structure analysis. AB - Proteins tend to use recurrent structural motifs on all levels of organization. In this paper we first survey the topics of recurrent motifs on the local secondary structure level and on the global fold level. Then, we focus on the intermediate level which we call the short structural motifs. We were able to identify a set of structural building blocks that are very common in protein structure. We suggest that these building blocks can be used as an important link between the primary sequence and the tertiary structure. In this framework, we present our latest results on the structural variability of the extended strand motifs. We show that extended strands can be divided into three distinct structural classes, each with its own sequence specificity. Other approaches to the study of short structural motifs are reviewed. PMID- 8229096 TI - Boltzmann's principle, knowledge-based mean fields and protein folding. An approach to the computational determination of protein structures. AB - The data base of known protein structures contains a tremendous amount of information on protein-solvent systems. Boltzmann's principle enables the extraction of this information in the form of potentials of mean force. The resulting force field constitutes an energetic model for protein-solvent systems. We outline the basic physical principles of this approach to protein folding and summarize several techniques which are useful in the development of knowledge based force fields. Among the applications presented are the validation of experimentally determined protein structures, data base searches which aim at the identification of native-like sequence structure pairs, sequence structure alignments and the calculation of protein conformations from amino acid sequences. PMID- 8229097 TI - Preoperative HIV testing: is it less expensive than universal precautions? AB - Universal precautions are officially recommended to prevent HIV transmission in health care settings but for elective surgery some advocate routine preoperative HIV testing. These strategies have not been tested in clinical trials but universal precautions are very expensive and not cost-effective. Thus, for elective surgery, routine testing might save resources by permitting selective use of additional barrier precautions. We performed an economic evaluation to compare both strategies, using a simple approach to determine if routine testing (RT) is less expensive than universal precautions (UP). Conservatively assuming equal effectiveness in preventing HIV transmission, we compared a minimized estimate for the average cost of RT with a maximized estimate for the average cost of UP per elective operation. The minimized estimate for RT (US$57) was greater than the maximized estimate for UP (US$36) per procedure. Results were stable or strengthened by sensitivity analysis. Routine HIV testing is not a valid economic alternative to UP for elective surgery. The simple methodology used in this study can be a preliminary strategy to review other strategies for preventing HIV transmission. This method is particularly useful when data are inadequate for a formal economic evaluation to determine the utility of collecting the detailed information necessary for a full comparison. PMID- 8229098 TI - The case for a full cost-benefit analysis of preoperative HIV screening. PMID- 8229099 TI - Evidence-based vs emotion-based medical decision-making: routine preoperative HIV testing vs universal precautions. PMID- 8229100 TI - Thrombolytic therapy in acute myocardial infarction: some reservations. PMID- 8229101 TI - Knowledge of medical history information among proxy respondents for deceased study subjects. AB - Proxy respondents were interviewed for 96 decedents in an occupational cohort. A second respondent was interviewed for 59 decedents. Medical records were reviewed to validate questionnaire information. The percentage of respondents who answered "don't know" (non-response) to questions about medical condition ranged from 5% (cancer and heart disease) to 17% (ulcers). Non-response rates were lowest among spouses, intermediate among children, parents, and siblings, and highest among other relatives and friends. Among 41-55 pairs, depending on the condition, agreement between paired respondents was excellent (kappa > 0.75) for ulcers, cancer, diabetes, and lung disease. A higher percentage of medical records was obtained for decedents with spouse respondents and for decedents with more recent dates of death. Sixty percent or more of the medical records were obtained for patients with cancer (n = 30), heart disease (n = 26), stroke (n = 9), and liver disease (n = 10). The positive predictive value of the proxy respondent information for these conditions was 93, 81, 78, and 60%, respectively. PMID- 8229102 TI - A self-administered quality-of-life questionnaire after acute myocardial infarction. AB - A slightly modified version of the Quality-of-Life after Myocardial Infarction (QLMI) questionnaire developed by Oldridge and colleagues was applied in a self administered mode to patients with suspected acute myocardial infarction (AMI) in a randomized controlled trial of secondary prevention. Acceptability of the questionnaire was good, with 93% of responders answering all items. Factor analysis suggested three quality-of-life (QL) dimensions which we called "emotional", "physical" and "social". These differed somewhat from the dimensions proposed by Oldridge and colleagues. However, a sensitivity analysis showed relative invariance of results to weighting schemes. Scores on our three dimensions were responsive to differences between the treatment groups, and demonstrated construct validity based on associations between the measured QL and variables expected to affect QL. We conclude that the QLMI questionnaire has good potential as an instrument for assessing QL in post-AMI patients and that it can be successfully self-administered. PMID- 8229103 TI - Relationship between hemoglobin and cardiovascular risk factors in young adults. AB - To understand mechanisms of association between hemoglobin and cardiovascular disease (CVD), the relationships between hemoglobin and CVD risk factors were examined in 5115 black and white men and women who participated in the Coronary Artery Risk Development in Young Adults (CARDIA) Study. Hemoglobin was higher in men than women, whites than blacks, and smokers than non-smokers (p < 0.001). After adjusting for age, body mass index, current smoking status, and clinical center, hemoglobin correlated with diastolic blood pressure (0.11 < or = r < or = 0.22, p < 0.001) and plasma total cholesterol (0.08 < or = r < or = 0.11, p < 0.01) in all four race-sex groups and with systolic blood pressure in all but black women (0.07 < or = r < or = 0.13, p < 0.05). Among other factors possibly related to CVD risk, only serum albumin and white blood cell count showed significant correlations with hemoglobin in all groups (0.19 < or = r < or = 0.27, 0.07 < or = r < or = 0.18, respectively). These findings suggest that an association of hemoglobin with CVD risk factors may explain the association of hemoglobin with CVD. PMID- 8229104 TI - Segmental arterial disease in the lower extremities: correlates of disease and relationship to mortality. AB - The purpose of this study was to identify the correlates of disease and mortality risk associated with the presence of stenosis in the major arterial segments of the lower limb in men and women evaluated in a peripheral vascular laboratory. A total of 326 men and 249 women had unisegmental stenosis; 312 men and 275 women had multisegmental disease; 132 men and 111 women had no apparent disease. Multivariate analysis indicated that current smoking and elevated systolic blood pressure were the key risk factors associated with isolated aortoiliac and femoropopliteal arterial disease in both men and women. A history of diabetes was significantly associated with tibioperoneal disease in men while elevated systolic blood pressure was the major correlate of distal disease in women in this patient population. The relative risk of mortality was elevated 2- to 7-fold in men and women with multi- and unisegmental disease involving the aortoiliac and femoropopliteal segments; presence of tibioperoneal disease did not significantly increase mortality relative to those who were free of disease. These results suggest that the etiology and mortality risk associated with atherosclerosis in the lower extremity may vary with the anatomic site and/or severity of the lesion. PMID- 8229105 TI - Selection of neighborhood controls for a study of factors related to the diagnosis of cervical cancer. AB - The level of effort required to generate neighborhood controls for a statewide matched case-control study of cervical cancer was investigated, with the aim of identifying hard-to-reach demographic subgroups. Cross reference telephone directories were used to identify households on the same street as the case. Letters were then sent to the households, followed by 'phone calls. A total of 2920 households were contacted to obtain 147 controls. Overall, 63.6% of age eligible contacts participated in the study. In 49.3% of all households the major reason for not obtaining a control was "no age-eligible women". Level of effort required to obtain a matched control was greater for black women than for white women--on average 24 letters and 40 'phone calls for black women vs 12 letters and 20 calls for white women. Fewer eligible younger women refused to be interviewed than older. No marked differences were noted when the data were stratified by urban-rural area of residence. PMID- 8229106 TI - Stone-age genes and modern lifestyle: evolutionary mismatch or differential survival bias. PMID- 8229107 TI - The Tromso Study: frequency and predicting factors of analgesic drug use in a free-living population (12-56 years). AB - In a cross-sectional survey carried out in Tromso in 1986-7, 19,137 men and women aged 12-56 years from the general population were asked about their use of drugs during the preceding 14 days. Use of analgesics was very common. On average 28% of the women and 13% of the men had used analgesics. Drug use due to menstruation discomfort contributed only partly to the gender difference. Drug use was independent of age from 20 years of age. Factors having an impact on analgesic drug use were analyzed by logistic regression. The most significant predictors of analgesic use were suffering from headache (OR = 14.2(women) OR = 24.4(men)) and infections (OR = 2.0(women) OR = 2.4(men)). Drug users also tended to suffer from symptoms of depression (women) and sleeplessness (men). Lifestyle and sociodemographic factors were also significant predictors, but were of marginal importance (OR < 1.5) compared with occurrence of pain and infections. PMID- 8229108 TI - The impact of age on risk of adverse drug reactions to digoxin. For The Gruppo Italiano di Farmacovigilanza nell' Anziano. AB - To assess the association of age and other potential risk factors with digoxin toxicity, adverse drug reactions to digoxin (ADRDIG) were studied in all patients (n = 1338) on digoxin therapy consecutively admitted to 41 clinical wards throughout Italy during 4 months in 1988. At the time of admission, 28 patients (2.1%) had evidence of ADRDIG. In multivariate logistic regression analysis, significant associations with ADRDIG were found for age > or = 80 years compared to age 65-79 years (OR = 2.75, 95% CI = 1.17-6.45), daily digoxin dosage of > or = 0.25 mg (OR = 2.51, 95% CI = 1.16-5.47), serum creatinine > or = 120 mumol/L (OR = 3.75, 95% CI = 1.69-8.32), and for treatment with amiodarone, propafenone, quinidine or verapamil (OR = 2.60, 95% CI = 1.07-6.30). Those aged < 65 years had a similar risk of digoxin toxicity as those aged 65-79 years (OR = 1.07, 95% CI = 0.28-4.12). Adverse drug reactions to digoxin were found in 1 in 50 patients hospitalized on digoxin therapy. Patients aged 65-79 years were not at increased risk for digoxin toxicity compared to younger patients, while advanced age (> or = 80 years) was an independent risk factor for this outcome. PMID- 8229109 TI - The use of tranquilizers, hypnotics and analgesics among 18,592 Finnish adults: associations with recent interpersonal conflicts at work or with a spouse. AB - The use of tranquilizers, hypnotics, and analgesics was assessed by a postal questionnaire in a nationwide sample of 18,592 adults aged from 24 to 65 years: 14,800 employees, 2105 housewives and 687 unemployed persons. The use of tranquilizers was most frequent among unemployed persons whereas housewives did not use the studied medicines more than the employed women. In the multivariate analyses (adjusted for age, social class and neuroticism) unmarried status was associated with the use of tranquilizers and hypnotics among both genders. Reported recent interpersonal conflicts both at work and with a spouse were associated with the use of tranquilizers among men, whereas among women significant associations were found only for conflicts with a spouse. These results indicate that domestic factors are more likely to predict women's tranquilizer use. PMID- 8229110 TI - Causality assessment of adverse reactions to drugs--I. A novel method based on the conclusions of international consensus meetings: application to drug-induced liver injuries. AB - Despite the great number of methods proposed, assessing the causal role of a drug in the occurrence of an adverse medical event remains one of the most controversial issues. Qualifying terms for criteria, such as "compatible", "suggestive" of "inconclusive", have never been strictly defined, leading to low reproducibility. Weights of the criteria are usually not adapted to the injured organ, decreasing the specificity of the method. In this paper, a new method for drug causality assessment is described. Contents and limits of the criteria have been defined by experts convened to organ-oriented international consensus meetings. Additional criteria have been introduced and weights attributed. The method was applied to reports of acute liver injuries. The reproducibility was tested by an independent team. The validity of this novel method is studied in the following paper, based on an original approach using reports with positive rechallenge as external standard. PMID- 8229111 TI - Causality assessment of adverse reactions to drugs--II. An original model for validation of drug causality assessment methods: case reports with positive rechallenge. AB - Standards are lacking for validation of drug causality assessment methods. An original model is proposed using a positive rechallenge as an external standard. This model was used to validate the novel causality assessment method (RUCAM) described in the previous article (Part I; J Clin Epidemiol 1993; 46: 1323). Seventy seven reports of drug-induced acute liver injuries with positive rechallenge were collected from the medical literature and divided into 49 cases and 28 controls. The RUCAM was applied to information obtained prior to readministration. The score was significantly higher (p < 10(-4)) in cases than in controls with high levels of sensitivity, specificity and predictive values. It is concluded that (1) adverse drug reaction reports with a positive rechallenge can provide a standard for validation of causality assessment methods, (2) RUCAM applied to drug-induced liver injuries has been validated. PMID- 8229112 TI - Multiple sclerosis: stressors and coping strategies in spousal caregivers. AB - Community health nurses, who have frequent and prolonged contact with patients afflicted with chronic disease, play a central role in the assessment and care of patients and their families who must deal with the many ramifications of long term disease. The importance of multiple sclerosis in health care is a result of its worldwide prevalence rate of 57.9 per 100,000 population, its tendency to occur in the young adult, and its chronicity. This exploratory study was designed to describe the stressors of caregivers of individuals with multiple sclerosis, to identify coping behaviors used by caregivers, and to explore the relation between caregiver stress and caregiver coping behavior. The study sample included 20 caregiving spouses of individuals with multiple sclerosis. The data were collected through a semistructured interview of caregiving spouses. Caregiver coping behavior was measured with the Ways of Coping Checklist (Folkman & Lazarus, 1980), and caregiver stress was measured with the Caregiver Strain Index (B. C. Robinson, 1983). Descriptive statistics were calculated for each of the demographic and disability variables. Pearson product-moment correlations were employed to assess the relation between caregiver coping behavior and caregiver stress. Findings revealed significant correlations between caregiver stress and caregiver coping behavior. These findings indicate that as stress in the caregiving role increases, there is an increase in the use of various forms of coping behaviors, both problem focused and emotion focused. Types of stressors and coping strategies used by caregivers are described, and nursing implications are discussed. PMID- 8229113 TI - Information transmission in bilingual, bicultural contexts: a field study of community health nurses and interpreters. AB - The purpose of this article is to present the findings made in a field study of nurses and interpreters who were working in a county health department. Informed by the qualitative research method of grounded theory, analysis of 37 semistructured interviews revealed three types of interactions among nurses, interpreters, and clients. In these situations, the interpreter worked as a voice box, an excluder, or a collaborator. Collaborative interactions allowed both interpreters and community health nurses to establish the most effective rapport with clients. Rapport, in turn, facilitated client assessment and the planning and delivery of nursing services. Implications for community health nursing practice include providing opportunities for interpreters to establish a rapport with clients. Research implications include the development of strategies that promote collaboration between community health nurses and interpreters. Theoretically, such strategies would enhance client outcomes. PMID- 8229114 TI - Managing uncompensated home care. AB - The purpose of this study was to determine the strategies used by managers of public health nursing and visiting nurse associations to cope with increased demand for uncompensated home care. A questionnaire was mailed to all public health nursing and visiting nurse associations in the upper midwest. Of the 161 agencies reporting their experience with uncompensated care demand, 129 experienced an increased demand for uncompensated care. Public health nursing and small agencies were more likely to experience increased demand for uncompensated care. Use of cost reduction, revenue enhancement, and quality assurance strategies varied by demand for uncompensated care status as well as by agency size and type. Managers of large and medium agencies and visiting nurses associations were more likely to use the identified strategies. Six strategies were used by more than 50% of the responding agencies. PMID- 8229115 TI - Types of health problems and satisfaction with services in a rural nurse-managed clinic. AB - The purpose of this study was to investigate the satisfaction of clients with care provided in a rural nurse-managed health center by family nurse practitioners and to describe common health problems for which family nurse practitioner services were sought. The results indicated that of the 2,106 visits from a case load of 1,350 clients, acute health problems were the most common (95.7%), and chronic health problems were the least common (3.9%). Respiratory and ear infections were the most frequent acute illness (55%), and hypertension (29%), respiratory (27%), and musculoskeletal complaints (27%) were the most frequent chronic conditions. Children up to 6 years of age constituted the largest client population (30%), followed by young adults (23%). Older adults were the smallest numbers of clients (3%). Client satisfaction with nurse practitioner-managed care was highly rated by 97% (n = 101) of the subjects in a random sample telephone survey. The results suggested that family nurse practitioners were providing effective and satisfactory healthcare for this rural community. Issues for further investigation are also discussed. PMID- 8229116 TI - Patient education: bridging the gap between inpatient and ambulatory care. AB - This article describes a model curriculum of perinatal education designed to bridge the gap between prenatal ambulatory care education and labor and delivery, postpartum and well child care in a high-risk population of inner city women. The curriculum, which consists of broad-based, maternal-child health information, was developed, implemented, and evaluated through an interdisciplinary approach. The initiative narrowed the preexisting gap between departments, increased clients' knowledge, and promoted positive changes in clients' behavior. The breadth of the curriculum content addresses the unquestionable need for perinatal education that will enhance compliance and empower clients toward health maintenance. PMID- 8229117 TI - When the population is a congregation: the emerging role of the parish nurse. PMID- 8229118 TI - Interleukin-3: a potentially useful agent for treating chemotherapy-related thrombocytopenia. PMID- 8229119 TI - Dietary fat reduction among women with early breast cancer. PMID- 8229120 TI - Dose-dependent interleukin-3 stimulation of thrombopoiesis and neutropoiesis in patients with small-cell lung carcinoma before and following chemotherapy: a placebo-controlled randomized phase Ib study. AB - PURPOSE: To evaluate the safety, tolerance, and hematologic effects of recombinant human interleukin-3 (IL-3) in patients with small-cell lung cancer (SCLC) before and following multiagent antineoplastic therapy in a placebo controlled, randomized, double-blind study. PATIENTS AND METHODS: Twenty-eight patients (22 men and six women; median age, 60 years) with previously untreated SCLC entered the study. Patients were assigned to six groups of escalating-dose IL-3 ranging from 0.25 to 10 micrograms/kg/d administered by continuous infusion for 7 days, with one patient in each group receiving placebo. After a 1-week interval, the first of three cycles of carboplatin, etoposide (VP16), and epirubicin (CVE) given every 3 weeks was administered. The second cycle of CVE was followed by 7 days of IL-3 administered at the same daily dose as administered during the first infusion. RESULTS: The maximum-tolerated dose was not encountered in this study. Fever was the most frequently observed side effect. Before any chemotherapy, World Health Organization (WHO) grade II fever only appeared at doses > or = 2.5 micrograms/kg/d. Other side effects included rash, headache, and myalgia. During the first infusion of IL-3, before administration of chemotherapy, dose-dependent increases in peripheral-platelet counts (r = .613; P < .001) and neutrophil counts (r = .505; P = .007) were observed. Following the second cycle of CVE, recovery of peripheral platelet counts was faster as compared with the first cycle of CVE for patients treated with 7.5 and 10 micrograms/kg of IL-3 (P = .021). Chemotherapy postponements due to myelotoxicity were also less frequent following the second cycle of CVE as compared with the first for patients treated with > or = 2.5 micrograms/kg of IL 3 (P = .036). Compared with an age-matched historical group receiving identical chemotherapy (n = 191), administration of IL-3 did not modify either disease-free survival or overall patient survival rates. CONCLUSION: IL-3 is well tolerated at doses up to 10 micrograms/kg/d. In the absence of chemotherapy, biologic effects on both neutrophils and platelets were seen at doses > or = 2.5 micrograms/kg/d. IL-3 infusion following the second cycle of CVE appears to reduce chemotherapy induced myelosuppression, but does not alter tumor response or patient survival rates. PMID- 8229121 TI - Adherence to a dietary fat intake reduction program in postmenopausal women receiving therapy for early breast cancer. The Women's Intervention Nutrition Study. AB - PURPOSE: To evaluate the feasibility of integrating a program based on dietary fat intake reduction into adjuvant treatment strategies for postmenopausal women receiving therapy for early breast cancer. PATIENTS AND METHODS: Two hundred ninety postmenopausal women with localized (stage I to IIIa) breast cancer receiving conventional systemic therapy provided informed consent and were randomized in a multicenter trial to either a dietary intervention group receiving a program of individualized instruction for reducing total fat intake or a dietary control group with minimal dietary counseling. RESULTS: Significantly reduced (P < .001) fat intake (in terms of percent calories derived from fat) was observed in the intervention group versus the control group at 3 months (20.3% +/- 2.4% v 31.5% +/- 2.6%, mean +/- SD, respectively) and maintained throughout 24 months of observation. The 50% reduction in daily fat gram intake (from 66 +/- 23 to 33 +/- 14 g, P < .001) seen at 6 months was associated with reduced saturated fat, monounsaturated fat, polyunsaturated fat, and linoleic acid (P < .001). Significantly lower body weight was also seen in intervention compared with control patients at all observation periods, resulting in a 3.3-kg weight difference 18 months after randomization (P < .001). CONCLUSION: Substantial and sustained dietary fat reduction with associated weight change can be achieved at relatively low cost within the context of conventional multimodality clinical management of postmenopausal women with localized breast cancer. This result supports the feasibility of conducting a full-scale evaluation of the influence of dietary fat intake reduction on the clinical outcome of breast cancer patients. PMID- 8229122 TI - Effect of megestrol acetate on quality of life in a dose-response trial in women with advanced breast cancer. The Cancer and Leukemia Group B. AB - PURPOSE: The impact of the side effects of megestrol acetate on the quality of life of noncachectic women with advanced breast cancer was studied in a dose response clinical trial of the Cancer and Leukemia Group B (CALGB 8741). Side effects of appetite increase and weight gain at higher doses were predicted to have a negative effect on quality of life. PATIENTS AND METHODS: Stage IV breast cancer patients were randomized to receive either 160, 800, or 1,600 mg/d of megestrol acetate. Quality of life was assessed in 131 patients at trial entry and at 1 and 3 months while on treatment, by telephone interview, using the following measures: the Functional Living Index-Cancer (FLIC), Rand Functional Limitations Scale, Rand Mental Health Inventory (MHI), the Body Image Subscale, and linear analog scales of drug side effects. RESULTS: At 3 months, women treated with 160 mg/d reported less severe side effects (P < .0005), better physical functioning (FLS, P < .0005), less psychologic distress (MHI, P = .008), and an improvement in overall quality of life (FLIC, P = .003) from the time of study entry as compared with those treated with 1,600 mg/d. Patients who received the 800-mg/d dose fell between the low- and high-dose arms in reported intensity of drug side effects, but responded similarly to those in the 160-mg/d group in terms of physical functioning, psychologic distress, and overall quality of life. CONCLUSION: Unless additional follow-up data demonstrate a survival advantage at higher doses, the 160-mg/d dose is optimal, achieving maximal treatment effect with the fewest side effects and better quality of life. PMID- 8229123 TI - Factors influencing prognosis in node-negative breast carcinoma: analysis of 767 T1N0M0/T2N0M0 patients with long-term follow-up. AB - PURPOSE: This study was undertaken to define prognostically favorable and unfavorable subgroups of node-negative breast carcinoma patients by employing conventional pathologic data. PATIENTS AND METHODS: Seven hundred sixty-seven women with T1N0M0/T2N0M0 breast carcinoma treated consecutively from 1964 through 1970 by modified or radical mastectomy without systemic adjuvant therapy were analyzed at a median follow-up duration of 18 years. RESULTS: Size and histologic type of the carcinoma were crucial discriminants of prognosis. We defined a prognostically favorable group of 219 patients (29%) with infiltrating duct or lobular carcinoma < or = 1.0 cm in diameter or special tumor types < or = 3.0 cm. This group had a relapse-free survival rate of 91% at 10 years and 87% at 20 years. The less favorable group (548 patients, 71%) with infiltrating duct or lobular carcinoma greater than 1.0 cm and special tumor types greater than 3.0 cm had relapse-free survival rates of 73% and 68% at 10 and 20 years, respectively. The frequency of nonmammary malignant neoplasms (NMMN) was similar to that of contralateral carcinoma. Deaths due to NMMN were seven times more frequent than deaths due to contralateral carcinoma. CONCLUSION: Nearly 30% of these node negative patients, identified on the basis of tumor size and type, had an extremely favorable prognosis. There is insufficient evidence to warrant the routine use of adjuvant therapy in this group unless new forms of treatment prove to be less toxic and/or more effective in enhancing relapse-free survival. Early detection of NMMN should be an important part of the follow-up of node-negative breast carcinoma patients. PMID- 8229124 TI - Metabolic monitoring of breast cancer chemohormonotherapy using positron emission tomography: initial evaluation. AB - PURPOSE: We assessed the feasibility of noninvasive metabolic monitoring of cancer chemohormonotherapy using sequential quantitative positron emission tomographic (PET) scans of tumor glucose metabolism with the glucose analog 2 [18F]-fluoro-2-deoxy-D-glucose (FDG). PATIENTS AND METHODS: Eleven women with newly diagnosed primary breast cancers larger than 3 cm in diameter beginning a chemohormonotherapy program underwent a baseline and four follow-up quantitative PET scans during the first three cycles of treatment (days 0 to 63). Tumor response was sequentially determined clinically, radiographically, and then pathologically after nine treatment cycles. RESULTS: Eight patients had partial or complete pathologic responses. Their maximal tumor uptake of FDG assessed by PET decreased promptly with treatment to the following: day 8, 78 +/- 9.2% (P < .03); day 21, 68.1 +/- 7.5% (P < .025); day 42, 60 +/- 5.1% (P < .001); day 63, 52.4 +/- 4.4% (P < .0001) of the basal values. Tumor diameter did not decrease significantly during this period through 63 days. Prompt decreases in the FDG influx rate (K) from basal levels (from .019 to .014 mL/cm3/min) after 8 days of treatment (P < .02) and in the estimated rate of FDG phosphorylation to FDG-6 phosphate (k3) from .055 to .038 min-1 after 8 days of treatment (P < .02) to .029 +/- .004 min-1 at 21 days) (P < .02) were observed. Three nonresponding patients had no significant decrease in tumor uptake of FDG (81 +/- 18% of basal value), influx rate (.015 to .012 mL/cm3/min), or tumor size (81 +/- 12% of basal diameter) comparing basal versus 63-day posttreatment values. CONCLUSION: Quantitative FDG PET scans of primary breast cancers showed a rapid and significant decrease in tumor glucose metabolism after effective treatment was initiated, with the reduction in metabolism antedating any decrement in tumor size. No significant decrease in FDG uptake (SUV) after three cycles of treatment was observed in the nonresponding patients. FDG PET scanning has substantial promise as an early noninvasive metabolic marker of the efficacy of cancer treatment. PMID- 8229125 TI - Carboplatin in the treatment of advanced breast cancer: a phase II study using a pharmacokinetically guided dose schedule. AB - PURPOSE: We performed a phase II study of single-agent carboplatin against metastatic/locally advanced breast cancer using a pharmacokinetically guided dose schedule, to define further the potential role for this agent in combination and high-dose therapy. PATIENTS AND METHODS: Forty patients with metastatic/locally advanced breast cancer were treated with carboplatin based on glomerular filtration rate (GFR) to achieve an area under the concentration-versus-time curve (AUC) of 7 mg/mL.min, with each course repeated at 4-week intervals. The median age was 57 years (range, 37 to 71). RESULTS: Ten patients achieved a partial response (PR), for an overall response rate of 25% (95% confidence interval, 13% to 41%). One of 13 (8%) previously treated patients responded compared with nine of 27 (33%) patients who had not received previous chemotherapy. Median response duration was 18 weeks (range, 10 to 68). World Health Organization (WHO) grade 2 or greater toxicity was as follows: anemia, 42%; leukopenia, 20%; thrombocytopenia, 35%; nausea/vomiting, 39%; and infection, 9%. CONCLUSION: This study confirms other reports indicating that carboplatin has moderate activity in previously untreated patients, but not in previously treated patients. In our view, carboplatin is a more appropriate agent than cisplatin for inclusion in high-dose chemotherapy schedules with autologous bone marrow rescue, and our results support the concept of calculating dose escalation on the basis of the area under the dose-response curve using the Calvert formula, rather than on surface area. PMID- 8229126 TI - High-dose carboplatin and recombinant granulocyte-macrophage colony-stimulating factor in advanced-stage recurrent ovarian cancer. AB - PURPOSE: We investigated whether carboplatin myelosuppression could be favorably modulated by the administration of recombinant granulocyte-macrophage colony stimulating factor (rGM-CSF) in patients with advanced-stage ovarian cancer. PATIENTS AND METHODS: Thirty-four patients with advanced-stage recurrent ovarian cancer were treated with high-dose carboplatin (800 mg/m2 per 35-day cycle) and rGM-CSF. rGM-CSF was administered as a daily subcutaneous injection starting 72 hours after the carboplatin dose and continuing until 7 days beyond the WBC nadir. rGM-CSF was administered in a phase I fashion. Seven patients were treated at an rGM-CSF dose of 3 micrograms/kg, 11 at 5 micrograms/kg, 10 at 10 micrograms/kg, and six at 20 micrograms/kg. RESULTS: rGM-CSF-related toxicities that were not dose-related included nonneutropenic fever, rib pain, acute hypersensitivity reaction, and pericarditis. At the rGM-CSF dose of 20 micrograms/kg, debilitating malaise was seen in four of six patients and this was the dose-limiting toxicity. Patient tolerance of the 3-micrograms/kg and 5 micrograms/kg doses was good, but tolerance was limited for the 10-micrograms/kg dose. Febrile neutropenia was seen in four of seven patients at 3 micrograms/kg, two of 11 at 5 micrograms/kg, two of 10 at 10 micrograms/kg, and one of six at 20 micrograms/kg. Cumulative carboplatin myelotoxicity was blunted only in respect to WBC count, and not for platelets or RBCs. Gastrointestinal bleeding was seen in seven patients. The administered dose-intensity of carboplatin averaged 134 mg/m2/wk for the cohort, or 670 mg/m2 per 35-day cycle. There were two clinical complete responses and eleven partial responses, for a response rate of 38%. CONCLUSION: rGM-CSF appears to be effective and tolerable at 5 micrograms/kg/d administered subcutaneously, if given with carboplatin doses up to approximately 600 mg/m2 over 35 days. The use of rGM-CSF with high-dose carboplatin is associated with a substantial response rate in poor-prognosis ovarian cancer patients. PMID- 8229127 TI - Concurrent radiation therapy and chemotherapy for locally unresectable squamous cell head and neck cancer: an Eastern Cooperative Oncology Group pilot study. AB - PURPOSE: The feasibility and success of an intensive chemoradiotherapeutic protocol for patients with locally advanced, unresectable squamous cell head and neck cancer was tested in this limited-institution, Eastern Cooperative Oncology Group phase II pilot study. MATERIALS AND METHODS: Between December 1987 and September 1989, 57 patients were entered onto this trial. The treatment protocol consisted of three courses of a 4-day continuous fluorouracil infusion, a single cisplatin bolus injection, and concurrent split-course radiotherapy. After 30 Gy of radiation and two chemotherapy courses, patients were evaluated for response and for the possibility of surgical resection. RESULTS: Fifty-five of 57 registered patients are assessable for toxicity and 52 are assessable for response and survival. Toxicity was significant, but tolerable, although there were three toxic deaths. A complete response to this treatment was ultimately achieved by 77% of patients. Twenty-four patients remain relapse-free. The projected Kaplan-Meier 4-year relapse-free survival rate is 45% and the overall survival rate is 49%. Median relapse-free and overall survival durations are 26 and 37 months, respectively. Of the 28 treatment failures, 79% were locoregional. Fourteen patients underwent surgery. Six remain relapse-free. CONCLUSION: This aggressive concurrent chemoradiotherapy protocol appears feasible within a cooperative group. Treatment results are promising and appear durable. A randomized phase III clinical trial is currently underway. PMID- 8229128 TI - Clinical implications of the serum level of CD23 in patients with undifferentiated nasopharyngeal carcinoma. AB - PURPOSE: In contrast with other carcinoma cells, cells from nude mice transplanted undifferentiated carcinoma of nasopharyngeal type (UCNT) release the soluble fragment of the CD23 antigen (sCD23). We sought to study the level of sCD23 in sera of untreated UCNT patients. PATIENTS AND METHODS: Pretherapeutic sera from 65 consecutive, locally advanced, initially nonmetastatic UCNT patients were assayed for sCD23. Patients were treated with a neoadjuvant chemotherapy/full-dose radiotherapy sequence. The mean follow-up duration is 50.5 months (range, 28 to 77). The Cox proportional hazards model was used to study the association between sCD23 levels and clinical signs and disease evolution. RESULTS: sCD23 levels showed an association with disease-free survival (DFS; P = .08) and overall survival (OVS; P = .08). Patients with sCD23 levels greater than a cutoff value of 0.6 ng/mL (greater cutoffs were found to be equally significant, but less sensitive), have a relative risk (RR) of relapse of 3.3 (95% confidence interval, 1.6 to 6.9; P = .002), and an RR of death of 2.9 (95% confidence interval, 1.2 to 7.3; P = .02), when taking other prognostic factors into account. CD23 does not correlate with either the response to treatment or the development of metastases, but appears to be related to local control (cutoff, 0.6 ng/mL; RR = 5.1 [95% confidence interval, 1.2 to 21.7]; P = .02). CONCLUSION: The serum level of sCD23 appears to be an independent prognostic factor for initially nonmetastatic, locally advanced UCNT patients, treated with chemotherapy and radiotherapy. Our data indicate an association between this marker and local relapses. Thus, a simple enzyme-linked immunoadsorbent assay (ELISA) could help to identify a high-risk group among nonmetastatic UCNT patients. CD23 could be a marker for two groups of UCNT tumors, with distinct biologic characteristics and clinical behaviors. PMID- 8229129 TI - Combined radiation and chemotherapy for invasive transitional-cell carcinoma of the bladder: a prospective study. AB - PURPOSE: To improve the results obtained by cystectomy alone and to determine the possibilities of conservative treatment in invasive bladder cancer, we designed a prospective study using a combination of fluorouracil (5-FU) plus cisplatin and concomitant radiation therapy, followed by either cystectomy or additional chemoradiotherapy. PATIENTS AND METHODS: Fifty-four patients with stage T2 to T4 operable untreated invasive bladder cancer were entered onto the study. Treatment was begun in all patients by transurethral resection (TUR) and followed by the 5 FU-cisplatin combination with concomitant bifractionated split-course radiation therapy. A control cystoscopy was performed 6 weeks after completion of the neoadjuvant program. Patients with persistent tumor underwent cystectomy. Complete responders were treated by either additional chemoradiotherapy (group A) or cystectomy (group B). RESULTS: At control cystoscopy, 40 of 54 patients (74%) had a histologically documented complete response. Four responders developed recurrent pelvic disease after a mean follow-up time of 27 +/- 12 months (three in group A and one in group B). Metastatic disease, which developed in 16 patients, occurred more frequently in the nonresponders (71%) than in responders (15%). The disease-free survival rate at 3 years was 62%; it was significantly better in responders (77%) than in nonresponders (23%). There was no difference in survival between groups A and B. CONCLUSION: This neoadjuvant chemoradiotherapy combination, easy to implement and well tolerated even in elderly patients, provides a high complete response rate. It may prove to be effective in inoperable patients and may be proposed as conservative treatment in patients with a complete response to the initial course of chemoradiation. PMID- 8229130 TI - Importance of continued testicular suppression in hormone-refractory prostate cancer. AB - PURPOSE: Patients in whom prostate cancer progresses despite testicular androgen ablation are generally said to have cancers that have become resistant to hormonal maneuvers. If androgen suppression has been pharmacologic, this therapy is often stopped before consideration of other systemic treatments. This exploratory study sought clinical correlates of experimental evidence that there may be substantial acceleration of tumor growth after cessation of androgen suppression. MATERIALS AND METHODS: A retrospective multivariate analysis was performed on survival data for 341 patients treated on four clinical trials of secondary therapy for hormone-refractory prostate cancer. Factors included in the model were recent weight loss, age, performance status, disease site (soft tissue v bone-dominant), prior radiotherapy, and continued androgen suppression v discontinued exogenous endocrine therapy. RESULTS: Recent weight loss, age, performance status, and disease site were important prognostic factors for survival duration in hormone-refractory prostate cancer. Correcting for these factors, continued testicular androgen suppression was also an important predictor of survival duration in all data sets examined. CONCLUSION: This retrospective study showed a modest advantage in survival duration for men with hormone-refractory prostate cancer who continued to receive testicular androgen suppression. The hypothesis that continued hormonal maneuvers can still affect survival in this group warrants examination in prospective trials. PMID- 8229131 TI - Sequential chemoimmunotherapy with cisplatin, interleukin-2, and interferon alfa 2a for metastatic melanoma. AB - PURPOSE: To evaluate the activity and the toxicity of the combination of cisplatin (CDDP)/recombinant interleukin-2 (rIL-2) and interferon alfa-2a (IFN alpha) in disseminated malignant melanoma (DMM). PATIENTS AND METHODS: Between December 1990 and March 1992, 39 patients with biopsy-proven metastatic malignant melanoma (MM), bidimensionally measurable lesions and an Eastern Cooperative Oncology Group (ECOG) performance status < or = 2 entered this protocol. Seventy nine percent had received previous chemotherapy including platinum complex (15%) and alpha interferon (44%). They received CDDP (100 mg/m2 on day 0) followed by IL-2 18.10(6) IU/m2/d continuous intravenous (IV) infusion from day 3 to day 6 and from day 17 to day 21. The cycle was repeated on day 28. Subcutaneous IFN alpha 9.10(6) IU three times weekly was administered throughout the treatment period. From day 66 or 94, patients were administered a maintenance cycle with CDDP 100 mg/m2, subcutaneous IL-2 5.10(6) IU/m2/d from day 15 to day 19 and from day 22 to day 26 and IFN alpha 9.10(6) IU three times weekly repeated every 5 weeks (maximum four cycles). RESULTS: Among 39 assessable patients, five patients achieved complete responses (CRs). Sixteen patients had partial responses (PRs). The overall objective response rate was 53.8%. The number of metastatic sites was the only response-predictive factor. Toxicity was manageable in a routine patient setting and there was no life-threatening toxicity. CONCLUSION: These results seem to indicate a possible synergy between CDDP/rIL-2 and IFN alpha in MM. PMID- 8229132 TI - A new peripherally implanted subcutaneous permanent central venous access device for patients requiring chemotherapy. AB - PURPOSE: A peripherally implanted central venous access device (P.A.S. Port; Pharmacia Deltec Inc, St Paul, MN) was evaluated for ease of insertion, functionality, acceptance, and complications in patients who required long-term venous access. A hand-held tracking system (Cath-Finder; Pharmacia Deltec Inc) used to determine catheter tip location was also evaluated. PATIENTS AND METHODS: A P.A.S. Port was placed in 47 patients who required long-term intravenous access. The median follow-up duration has been 32 weeks (range, 2 to 112). Total usage has been 2,028 catheter-weeks. The Cath-Finder was used to determine catheter tip location during insertion. Nursing staff and patient satisfaction were polled and functionality and complications were recorded. RESULTS: The device was found to be simple to insert, the procedure well tolerated, and, with one exception, the Cath-Finder accurately predicted catheter tip location. There was a 6.4% incidence of transient phlebitis and a 6.4% incidence of symptomatic axillary or subclavian vein thrombosis. There were no infectious complications. Access was simple in all but two obese patients. The device functioned well in all patients, except three in whom blood aspiration was difficult and two in whom fluid administration was slow. The device was well tolerated by all patients and nursing staff satisfaction was high. CONCLUSION: This device provides a highly acceptable, additional method of implantable, permanent central venous access for chemotherapy patients with a low complication rate. The successful use of the Cath-Finder and minor extent of the procedure may allow this device to be inserted in a clinic procedure room without sedation and fluoroscopy. PMID- 8229133 TI - Extended CSF cytarabine exposure following intrathecal administration of DTC 101. AB - PURPOSE: The aims of the present study were to determine ventricular and lumbar CSF pharmacokinetics and the maximum-tolerated dose (MTD) of DTC 101 (DepoFoam; DepoTech Corp, La Jolla, CA) following intraventricular administration. PATIENTS AND METHODS: Twelve patients with neoplastic meningitis were treated with escalating doses of DTC 101. CSF samples were obtained from the right lateral ventricle or from the lumbar subarachnoid space and cytarabine concentrations were determined by high-performance liquid chromatography. RESULTS: Therapeutic ventricular CSF concentrations were maintained for 9 +/- 2 days following administration of a single dose of DTC 101 into the lateral ventricle. Lumbar cytarabine concentrations became equal to those in the ventricle within the first 6 hours after intraventricular injection, and the subsequent decay in concentrations of free and total cytarabine were the same at both sites. Following intralumbar administration, the peak ventricular concentration of free cytarabine was reached within 1 day, and therapeutic ventricular CSF levels were maintained for several days. Therapeutic intralumbar concentration of free cytarabine was maintained for up to 14 days. The MTD was 75 mg of DTC 101, and seven of nine patients manifested cytologic responses. CONCLUSION: Extended CSF exposure to therapeutic cytarabine concentrations was achieved after a single intraventricular or intralumbar dose of DTC 101, permitting drug administration once every 2 weeks. PMID- 8229134 TI - Phase I and pharmacokinetic trial of weekly CPT-11. AB - PURPOSE: We conducted a phase I and pharmacokinetic trial of CPT-11 (irinotecan) to characterize the maximum-tolerated dose (MTD), toxicities, pharmacokinetic profile, and antitumor effects in patients with refractory solid malignancies. PATIENTS AND METHODS: We treated 32 patients with CPT-11 administered as a 90 minute intravenous infusion every week for 4 consecutive weeks followed by a 2 week rest period. Dose levels ranged from 50 to 180 mg/m2/wk. We determined concentrations of the lactone (active) and total (lactone plus carboxylate) forms of CPT-11 and its metabolite, SN-38, in the plasma and urine of selected patients during and after drug infusion. RESULTS: Grade 4 diarrhea was the dose-limiting toxicity (DLT) at the 180-mg/m2/wk dose level. Other toxicities attributed to CPT 11 included dehydration, nausea, vomiting, and asthenia. Hematologic toxicity was mild in most patients. The terminal plasma half-life for CPT-11 (total) was 7.9 +/- 2.8 hours, for CPT-11 (lactone) 6.3 +/- 2.2 hours, for SN-38 (total) 13.0 +/- 5.8 hours, and for SN-38 (lactone) 11.5 +/- 3.8 hours. We observed significant correlations between drug dose and peak plasma concentration (Cpmax) and between drug dose and area under the concentration curve (AUC) for CPT-11, but not for SN 38. CONCLUSION: The MTD for CPT-11 in this patient population was 150 mg/m2/wk when administered on a weekly-times-four schedule repeated every 6 weeks. At dose levels greater than 150 mg/m2/wk, diarrhea is dose-limiting. PMID- 8229135 TI - Phase I clinical trial of recombinant human tumor necrosis factor in children with refractory solid tumors: a Pediatric Oncology Group study. AB - PURPOSE: A phase I study was undertaken to determine the toxicity and maximum tolerated dose (MTD) of recombinant human tumor necrosis factor (rTNF) in children. PATIENTS AND METHODS: Twenty-seven patients with recurrent or refractory solid tumors were enrolled on the study. rTNF was administered daily for 5 days by 30-minute intravenous (IV) infusion, and doses were escalated in cohorts of three to six patients. Courses were repeated after a 9-day rest period, if toxicity was tolerable. Daily doses ranged from 100 to 350 micrograms/m2. RESULTS: Most courses were associated with grade I/II fever, rigors, nausea, or vomiting. Three patients experienced moderate dyspnea that responded to supplemental oxygen. All abnormalities resolved on discontinuation of the infusion. One patient had a cardiac arrest 90 minutes after receiving the first dose of rTNF and died 10 days later of related complications. In two other patients, rTNF was discontinued due to persistent grade IV hypotension. Toxicities were not consistently related to dose and no cumulative effects were noted. The dose-limiting toxicity was transient hepatic dysfunction, which occurred in three of six patients receiving 350 micrograms/m2; this toxicity was rapidly reversed on discontinuation of the rTNF. One patient, whose non-Hodgkin's lymphoma had recurred after bone marrow transplantation, had a partial response. Disease was stabilized in two patients. CONCLUSION: We recommend that phase II testing proceed at a dose of 300 micrograms/m2/d on the schedule described. PMID- 8229136 TI - Relation of tumor-cell ploidy to survival in children with medulloblastoma. AB - PURPOSE: To assess the value of tumor-cell ploidy as a predictor of survival in medulloblastoma. PATIENTS AND METHODS: Ploidy determinations were based on the flow-cytometric analysis of cellular DNA content in fresh tumor specimens taken from 34 consecutively treated children with newly diagnosed medulloblastoma. Patients were assigned a high or low risk of failure depending on tumor size and invasiveness, and the presence or absence of metastatic disease. Treatment consisted of radiotherapy, with or without chemotherapy, according to institutional or cooperative group protocols. RESULTS: Univariate analysis of candidate prognostic factors showed that only tumor-cell ploidy and clinical risk group had a statistically significant influence on survival. Patients with hyperdiploid stem lines (n = 9) had significantly longer survival times (P = .04) than did those with diploid lines (n = 20). The estimated 5-year survival probabilities (+/- SE) for these two subgroups were 89% +/- 11% and 48% +/- 13%, respectively. Although clinical risk status (high v low) showed essentially the same predictive strength as ploidy, the two features identified largely nonoverlapping subgroups. Thus, within the clinical high-risk group, it was possible to distinguish hyperdiploid patients whose 5-year survival rate (83% +/- 15%) was comparable to that of patients with localized, low-risk tumors. CONCLUSION: This prospective study indicates that both ploidy and clinical risk group are important prognostic factors in medulloblastoma. Their combined use at diagnosis would distinguish patients who require more aggressive therapeutic intervention (diploid, clinical high-risk group) from those who could be expected to benefit most from standard treatment. PMID- 8229137 TI - With modern imaging techniques, is staging laparotomy necessary in pediatric Hodgkin's disease? A Pediatric Oncology Group study. AB - PURPOSE: To determine whether the information gained from staging laparotomy can be predicted by imaging and/or clinical factors in children with Hodgkin's disease. PATIENTS AND METHODS: Between 1986 and 1991, 216 consecutive pediatric patients with Hodgkin's disease underwent laparotomy and were treated on two concurrent protocols in a multiinstitutional cooperative group. All patients had computed tomography (CT) of the chest, abdomen, and pelvis. Clinical factors studied included sedimentation rate, B symptoms, histology, number and location of involved sites, sex, mediastinal involvement, and age. Pretreatment CTs were centrally reviewed in 88 cases for the presence and size of both supradiaphragmatic and infradiaphragmatic lymph nodes, intrinsic spleen lesions, and splenic size. Models were generated that were predictive of any abdominal disease, splenic involvement, extensive splenic involvement, and upstaging at the laparotomy. False-positive and false-negative rates were calculated. RESULTS: For the end point of any abdominal disease, a model based on B symptoms, histology, sedimentation rate, and number and location of involved sites was highly significant (P < .0001). However, the success in predicting abdominal disease in an individual patient was limited: false-negative rate, 26%; false-positive rate, 32%. Highly significant models based on clinical factors and/or radiographic findings were also generated to predict splenic involvement, extensive splenic involvement, and upstaging with laparotomy, but they also had high false-positive and false-negative rates. CONCLUSION: Laparotomy findings cannot be predicted accurately in the majority of patients based on knowledge of CT findings and clinical factors. PMID- 8229138 TI - Patterns of relapse after autologous purged bone marrow transplantation for neuroblastoma: a Childrens Cancer Group pilot study. AB - PURPOSE: The goal of this investigation was to determine if comparing sites of neuroblastoma at relapse after myeloablative chemoradiotherapy and purged autologous bone marrow transplantation (ABMT) with sites of disease at diagnosis and before ABMT could provide insight to the reasons for treatment failure. PATIENTS AND METHODS: Ninety-nine patients with high-risk neuroblastoma underwent ABMT after induction chemotherapy, surgery +/- local radiation (RT) and then myeloablative therapy with teniposide (or etoposide), melphalan, doxorubicin, cisplatin, and total-body irradiation (TBI). RESULTS: Forty-one of 84 assessable patients (15 toxic deaths) developed progressive disease 1 to 44 months after ABMT. The overall probability of relapse 36 months after ABMT was 49%. Tumor recurred in primary (n = 22), bone (n = 20), bone marrow (n = 18), lung (n = 3), and other sites (n = 9). Eight patients relapsed in the primary site alone, 14 in primary and distant sites, and 19 in distant sites only. Of 41 patients with progressive disease, 33 have died, with a median interval from relapse to death of 4 months. Both bone and bone marrow involvement at diagnosis correlated with specific relapse in that site (P < .05). Bone marrow tumor content at harvest greater than 0.1% also correlated with bone marrow relapse (P = .001). There was an association between incomplete resection of the primary tumor at diagnosis and relapse in that site (P = .06). CONCLUSION: Neuroblastoma normally recurs in multiple sites after ABMT, particularly in areas of previous disease. More intensive treatment to known areas of disease (aggressive early surgery, effective myeloablative consolidation therapy) and post-ABMT therapy for minimal residual disease should be studied for their potential to decrease the frequency of relapse. PMID- 8229139 TI - Improved therapy for children with acute lymphoblastic leukemia and unfavorable presenting features: a follow-up report of the Childrens Cancer Group Study CCG 106. AB - PURPOSE: On past Childrens Cancer Group (CCG) trials, children with acute lymphoblastic leukemia and unfavorable presenting features had obtained an event free survival (EFS) rate of no better than 50%. Following promising pilot experience, this study was conducted to determine the benefit and morbidity of two intensive experimental regimens, Reg A, based on the Berlin-Frankfurt-Munster (BFM) 1976 regimen, and Reg B, the New York regimen. PATIENTS AND METHODS: Between February 1983 and November 1984, 217 eligible children with acute lymphoblastic leukemia and unfavorable presenting features were entered and randomly assigned to receive Reg A, Reg B, or Reg C, the control regimen. Assignment to Reg C was halted in November 1984 after interim analyses showed an inferior outcome. Subsequently, between November 1984 and March 1987, an additional 328 patients were randomly allocated to receive Reg A or Reg B. RESULTS: The 7-year EFS rate was 63% (+/- 6%, 1 SD) for Reg A, 61% (+/- 6%) for Reg B, and 40% (+/- 6%) for Reg C (P < .006). The difference between Reg A or Reg B and Reg C remained greater than 20 percentage points for EFS at 7 years and 15 percentage points for survival. Relative to Reg C, patients on Reg A accrued 16.3 additional days of hospitalization on average and, on Reg B, 20.2 days. EFS and survival were similar on Reg A and Reg B, but Reg B required more days of parenteral therapy and greater exposure to anthracyclines and alkylating agents. CONCLUSION: Both Reg A and Reg B provided a better outcome than Reg C for children with acute lymphoblastic leukemia and unfavorable presenting features. Outcomes on Reg A and Reg B were similar. Use of the more effective but more toxic regimens resulted in 78 additional hospital days per relapse prevented on Reg A and 101 days on Reg B. The current CCG trial for this population builds on Reg A. PMID- 8229140 TI - Autologous bone marrow transplantation for pediatric Hodgkin's disease: a case matched comparison with adult patients by the European Bone Marrow Transplant Group Lymphoma Registry. AB - PURPOSE: We analyzed the outcome of autologous bone marrow transplantation (ABMT) in pediatric and adult patients with Hodgkin's disease (HD) by means of a case controlled study. PATIENTS AND METHODS: Eighty-one pediatric HD patients who had undergone ABMT and who had been reported to the European Bone Marrow Transplant Group (EBMTG) Lymphoma Registry were compared with a case-matched group of 81 adult patients who had undergone the same procedure. The case-matching was performed following selection of the main prognostic factors for progression-free survival (PFS) by multivariate analysis. RESULTS: The PFS rate was not significantly different between the groups (39% for the pediatric group v 48% for the adult group). The overall relapse/progression rates for the groups were 52% and 40%, respectively. This was not a statistically significant difference and the sites of relapse were similar. There was no significant difference in the incidence or causes of procedure-related morbidity or mortality of the two groups. CONCLUSION: This study suggests that patients with pediatric HD have the same outcome at ABMT as their adult counterparts; however, long-term complications will need to be carefully monitored. PMID- 8229141 TI - In search of an optimal regimen for elderly patients with advanced-stage diffuse large-cell lymphoma: results of a phase II study of P/DOCE chemotherapy. AB - PURPOSE: The results of a prospective, phase II trial of an 8-week treatment program consisting of epirubicin or doxorubicin, vincristine, cyclophosphamide, etoposide, and prednisone (P/DOCE) for elderly patients with advanced large-cell lymphoma are reported and compared with previous phase II studies conducted in similar patients at the same institution. PATIENTS AND METHODS: Between March 1988 and September 1991, 63 previously untreated patients aged 65 to 85 years (median, 75) with advanced-stage diffuse large-cell lymphoma, defined as Ann Arbor stage III or IV or stage I or II with B symptoms or bulky disease, were enrolled on a brief, 8-week protocol consisting of five outpatient chemotherapy treatments. RESULTS: The complete response (CR) rate was 62%. The treatment related mortality rate was 8%, the actuarial 4-year failure-free survival (FFS) rate was 41%, and the overall survival (OS) rate was 45%. These results were compared with two earlier, 12-week protocols, low-dose doxorubicin, cyclophosphamide, vincristine, bleomycin, and prednisone (LD-ACOB-B) and etoposide, doxorubicin, bleomycin, and prednisone (VABE), performed at the same center. There was no difference in outcome among the three regimens. If all 133 patients treated on any one of these three specially designed regimen for elderly patients are combined, the projected 5-year OS rate is 38%. CONCLUSION: The 8 week P/DOCE chemotherapy regimen is equal in efficacy and similar in toxicity to 3 months of chemotherapy administered on a weekly schedule and similar to the results reported in the literature for longer, anthracycline-based chemotherapy treatments. There does not appear to be any improvement in outcome from more protracted treatment programs compared with the 8-week P/DOCE protocol. PMID- 8229142 TI - Role of magnetic resonance imaging in predicting relapse in residual masses after treatment of lymphoma. AB - PURPOSE: This prospective study of patients treated at the Royal Marsden Hospital Lymphoma Unit was designed to evaluate the role of magnetic resonance imaging (MRI) in the assessment of residual masses evident on computed tomographic (CT) scanning following treatment of lymphoma. PATIENTS AND METHODS: All patients had MRI, gallium-67 single-photon emission CT (67Ga SPECT), and erythrocyte sedimentation rate (ESR) performed within 3 months of completing therapy. Patients were monitored for 1 year posttreatment and observed for signs of relapse. Investigation results were correlated with disease status, and the sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) calculated. Time-to-relapse curves were derived and the log-rank test used to determine whether patients with a positive result were more likely to have a relapse within the mass than those with a negative result. RESULTS: Thirty-four patients were studied, 14 of whom relapsed, 11 within the area of residual mass. Overall, MRI had a high specificity (90%), PPV (71%), and NPV (75%), but poor sensitivity (45%). The results for 67Ga SPECT were similar, apart from lower sensitivity (33%). ESR had inferior performance in predicting relapse compared with the other tests. MRI was the only investigation to show statistical significance (P = .14) in predicting relapse, and this was particularly evident in Hodgkin's lymphoma (P = .003). Combining results of 67Ga SPECT and MRI did not improve predictive power. CONCLUSION: These data demonstrate that MRI is a valuable tool in the setting of a residual mass after treatment, giving clinically useful prognostic information. 67Ga SPECT also has a role, but is less effective in predicting relapse than MRI. PMID- 8229143 TI - General information tapes inhibit recall of the cancer consultation. AB - PURPOSE: Studies of tape recordings of cancer consultations have produced conflicting results. At the same time, audiotapes containing general information about cancer are poorly evaluated and are distributed to patients in an ad hoc manner. We compared the effects of both interventions on patient satisfaction, psychologic adjustment, and recall of information following their first consultation with a medical oncologist. PATIENTS AND METHODS: Patients (n = 142) were randomized to receive (1) an audiotape of their consultation, (2) an audiotape describing cancer in general terms, or (3) no tape. Recall of information was assessed in a structured interview 4 to 20 days after the consultation. RESULTS: Satisfaction with the consultation increased linearly from no tape to general tape to consultation tape. Satisfaction with the tape itself was higher in patients who received the consultation tape (satisfaction score, 61%) compared with those who received the general tape (43%). Average recall for all groups was 6.4 of the 25 items of information presented, and 2.4 of the six points identified as particularly important by the oncologist. The consultation tape did not improve recall over the no tape control, but the general tape caused a decrease of almost two items in total recall. Spontaneous (ie, unprompted) recall was significantly poorer with the general information tape. Psychologic adjustment to cancer was unaffected. CONCLUSION: We conclude that individual audiotapes have a limited potential to increase recall of information from the oncology consultation. General information tapes about cancer appear to inhibit recall actively. PMID- 8229144 TI - Secondary acute myelogenous leukemia in children with Latin-American ancestry. PMID- 8229145 TI - Second tumors after radiation treatment of testicular germ cell tumors. PMID- 8229146 TI - Splenectomy and the risk of developing leukemia. PMID- 8229147 TI - Salmonella arizonae peritonitis secondary to ingestion of rattlesnake capsules for gastric cancer. PMID- 8229148 TI - Different kinetic of enzymatic inactivation of lincomycin and clindamycin in Staphylococcus aureus. AB - Lincosamide inactivation nucleotidylation (Lin) enzyme determined by the pBI109PGL plasmid of Staphylococcus epidermidis exhibits high level resistance to lincomycin but sensitivity to clindamycin by standard susceptibility methods. Substrate profile determination showed clindamycin to be a better substrate for the enzyme than lincomycin. In cultures of the plasmid-harboring strain, the level of clindamycin decreased below the inhibitory concentration in the first 4 hours of incubation but the level of lincomycin persisted longer. The initial extended inhibitory effect of clindamycin is due to better membrane penetrating ability, resulting in a higher intracellular concentration than that of lincomycin. Moreover, energy-dependent reduction in clindamycin uptake, probably due to active efflux of clindamycin but not of lincomycin, was observed. A therapeutic effect of clindamycin is not expected in infections caused by Lin producer strains because the bacteriostatic effect of the drug is rapidly eliminated after administration. PMID- 8229149 TI - In vivo potentiation of polymorphonuclear leukocyte function by ciprofloxacin. AB - Ciprofloxacin was administered to 10 healthy volunteers at a dose of 250 mg orally. Serum and polymorphonuclear leukocytes (PMNLs) were obtained from all subjects before the administration of the drug and 12 hours after the administration. In addition serum was obtained from all subjects at 24 and 48 hours after ciprofloxacin administration. All sera and PMNLs were used for the chemotactic, phagocytic and killing determinations of the PMNLs. The results demonstrated that serum obtained 12 hours after the administration of ciprofloxacin potentiates PMNL chemotactic activity (chemotactic index (CI) = 33.0 +/- 4.2 microns (means +/- S.D.)) as compared to the chemotactic activity generated by serum obtained prior to administration of the drug, CI = 20.4 +/- 4.4 microns (p < 0.01). Serum obtained 24 and 48 hours after ciprofloxacin administration did not stimulate PMNL function. The administration of ciprofloxacin did not have any direct influence on the PMNLs in terms of their chemotactic response. Furthermore, serum obtained after the administration of ciprofloxacin markedly enhanced PMNL phagocytosis and killing of all organisms tested. Ciprofloxacin also acted directly on the PMNLs and increased their bactericidal activity. These results demonstrate that ciprofloxacin potentiates PMNL function in vivo which may be of potential clinical benefit. PMID- 8229150 TI - Immunostimulation by clarithromycin in healthy volunteers and chronic bronchitis patients. AB - The immune response to infecting pathogens may be either enhanced or depressed by therapeutic antimicrobial agents. Some macrolides have been shown to enhance aspects of the immune response. This study evaluates the effects of clarithromycin, a new broad-spectrum macrolide antibiotic, on leukocyte function in both healthy volunteers (single 500 mg dose) and chronic bronchitis patients (500 mg b.i.d.) by testing blood samples collected at baseline and at 2, 4, 8 and 12 hours after clarithromycin administration. Clarithromycin did not affect leukocyte chemotaxis but did significantly increase (P < 0.01) phagocytosis (both frequency and index), intracellular killing and natural killer activity, in healthy volunteers. In patients, clarithromycin enhanced phagocytosis frequency, index and intracellular killing. Clarithromycin appears to enhance the human immune response; the mechanism, pharmacodynamics, and clinical significance of this enhancement remain unknown. PMID- 8229151 TI - Clinical pharmacokinetics of theophylline during co-treatment with ticarcillin plus clavulanic acid in patients suffering from acute exacerbation of chronic bronchitis. AB - The effect of ticarcillin/clavulanic acid, taken for 7 days as vials containing 1000 mg of ticarcillin and 200 mg of clavulanic acid twice daily intramuscularly, on the steady-state pharmacokinetics of theophylline was studied in 12 patients suffering from acute exacerbation of chronic obstructive pulmonary disease. Initially, patients were treated for four days with theophylline as sustained release formulation in the amount of 600 mg daily; on the last day, blood samples were taken for theophylline determination. Theophylline concentrations were measured serially for 12 hours by the method of polarized immunofluorescence (Abbott TDx system). Subsequently, while theophylline was continued at the same dosage, each patient received in addition ticarcillin/clavulanic acid vials every 12 hours. After seven days of this combined medication, the serial assays of plasma were repeated at the same time intervals as before. No influence of ticarcillin/clavulanic acid was detectable on the steady-state theophylline pharmacokinetics. It is concluded that both drugs can be administered concomitantly without any dosage adjustment of theophylline. PMID- 8229152 TI - Influence of adriamycin dose in the outcome of patients with osteosarcoma treated with multidrug neoadjuvant chemotherapy: results of two sequential studies. AB - The results of two sequential studies of neoadjuvant chemotherapy for osteosarcoma of the extremities performed at Rizzoli Institute between 1986 and 1991 in 228 patients are presented. In both studies preoperative chemotherapy consisted of two cycles of high dose methotrexate (HDMTX), cisplatinum (CDP) and adriamycin (ADM). Postoperatively the good responder patients were treated with the same drugs used before surgery while in the poor responder patients ifosfamide was added to these three drugs. The preoperative treatment was the same in both studies while after surgery in the second protocol either the cumulative dose of ADM (270 mg/m2 instead of 360 mg/m2) or the single dose per cycle of this drug (60 mg/m2 instead of 90 mg/m2) was reduced. These changes in the last protocol were done to reduce the cardiotoxicity of ADM that was high in the first study (2 deaths and 1 heart transplantation). Since in the last protocol--in comparison with the first protocol--after surgery chemotherapy was restarted earlier and ADM was administered not as a single drug but in combination with the CDP the dose intensity of ADM was unchanged while the dose intensity of MTX, CDP and ifosfamide was higher than in the first study. The preliminary results of the 84 patients treated in the second study show a 2-year disease free survival significantly lower than that achieved in the 144 patients treated in the first study (37/51--73% vs 123/144--85%: P < 0.008). In addition, even if in the last study there were no cases of clinical cardiotoxicity due to ADM, there was a significantly higher percentage of severe myelodepression that led to two deaths for infectious complications. These results suggest that in neoadjuvant treatment of osteosarcoma the total dose of ADM and/or the single dose per cycle of the same drug are an important determinant of outcome and that increasing the dose-intensity of less toxic but less active agents, MTX, CDP and ifosfamide, at the expense of the more active and more toxic agent, ADM, can lead to a poorer outcome without reducing toxicity. PMID- 8229153 TI - No advantages in the addition of ifosfamide and VP-16 to the standard four-drug regimen in the maintenance phase of neoadjuvant chemotherapy of Ewing's sarcoma of bone: results of two sequential studies. AB - Between January 1988 and December 1990, 74 patients with localized Ewing's sarcoma of bone were treated with a new protocol that consisted of an initial 6 week period of chemotherapy with vincristine (VCR), adriamycin (ADM) and cyclophosphamide (EDX) followed by local therapy and additional chemotherapy with the same drugs previously indicated plus ifosfamide and VP-16. The rationale for the addition of ifosfamide and VP-16 to the four drugs of the standard chemotherapy of this tumor was that this drug combination was previously very effective in the treatment of metastases from Ewing's sarcoma even in patients who did not respond to cyclophosphamide. As local treatment all patients were offered surgery, when feasible (70 cases). Forty-three patients accepted and 27 refused. These patients, as the 4 patients in whom surgery was not considered feasible, were treated with radiation therapy alone (50-60 Gy). In the remaining patients amputation was performed in 4 cases, rotationplasty in 3 and resection in 36. Where conservative surgery was marginal or intralesional (30 cases), radiotherapy at lower doses (40-45 Gy) was also delivered. At a mean follow-up of 3.5 years (2-7), 43 patients (58%) remained continuously disease-free and 31 relapsed (29 with metastases and 2 with both metastases and local recurrences). These results do not differ from those obtained at our Institution in 98 patients treated between 1983 and 1988 with a neoadjuvant protocol in which only VCR, ADM, EDX and dactinomycin (DAC) were used (3-year continuously disease-free survival (CDFS) respectively of 54% and 55%). Despite the fact that these results came from a nonrandomized study, the Authors conclude that the addition of ifosfamide and VP-16 to the four-drug standard regimen did not improve the outcome of the patients with Ewing's sarcoma of bone which remains a lethal disease in about 50% of the cases. These findings stress the need to find more effective chemotherapeutic regimens for the associated treatment of this tumor. PMID- 8229154 TI - Lymphoblastoid interferon in advanced breast cancer: a phase II study. AB - Thirty-two metastatic breast cancer patients entered a phase II study in order to evaluate the toxicity and the clinical activity of lymphoblastoid interferon (IFN). The treatment was divided into two phases, induction and maintenance. During the first, patients were submitted to lymphoblastoid IFN at the dose of 3 MU die intramuscularly for 4-8 weeks; during maintenance treatment IFN was given at the same dosage, intramuscularly, every other day until progression. Five patients with inadequate follow-up were excluded from disease evaluation, thus 27 patients were evaluable for response. No complete response was observed; one patient achieved a partial response and 15 patients disease stabilization. Median time to progression in patients with partial response or stable disease was 11 weeks (range 2-32). Five patients with soft tissue metastases were biopsied before and after 1-2 months of treatment in order to perform Labelling Index and hormonal receptor status determinations. No significant modification was observed. PMID- 8229155 TI - Cisplatin and 5-fluorouracil as induction chemotherapy followed by radiation therapy in metastatic squamous cell carcinoma of an unknown primary tumor localized to the neck. A phase II study. AB - 12 patients with metastatic squamous cell carcinoma of an unknown primary tumor localized to the neck were treated with induction chemotherapy and radiotherapy. Median age was 54 years (range, 39-68 years) and male to female ratio was 2:1. Five (42%) patients had N3 disease and 7 (58%) had N2 disease. Induction chemotherapy consisted of cisplatin 60 mg/m2, days 1 and 2 and 5-fluorouracil (5 FU), 500 mg/m2, days 3 and 4. Cycles were repeated every 4 weeks to a total of 3 cycles. After 4 weeks rest, patients underwent radiation therapy. It was given to all possible sites of the primary tumor (nasopharynx, pyriform sinus, and the base of the tongue). The radiation therapy dosage planned to the whole neck, nasopharynx, and supraclavicular area was 45 Gy in 24 daily fractions in 5 weeks, increasing to 60-70 Gy to the metastatic site. We observed 9 (75%) complete responses and 1 (8%) partial response. 7 (58%) patients are currently with no evidence of disease. Median survival time for all patients is 36 months (range, 14+ to 56 months), while median survival time for N3 patients was 32 months (range, 26-56 months). Toxicities were predominantly mild to moderate nausea and vomiting and mucositis. This combined modality appears to be effective and feasible in treatment of metastatic squamous cell carcinoma of an unknown primary tumor localized to the neck. PMID- 8229156 TI - Carboplatin and etoposide in patients with advanced gastric cancer. AB - Thirty-six patients with advanced gastric cancer were treated with combination chemotherapy following surgery. Chemotherapy consisted of carboplatin (CBDCA) 300 mg/m2, days 1-3, and etoposide (VP 16) 100 mg/m2, days 1-5. 10/36 (28%) patients had objective response. The median duration of response was 8 months. Currently 26/36 patients have shown progressive disease at a median of 4 months. Three patients remain progression-free 7, 14, and 15 months after study entry, respectively. 26/36 patients have died and 10/26 remain alive at a median time of 16 months. Estimated survival for all patients is 7 months. Two groups of toxicities were observed: hematological and gastrointestinal. They were generally assessed as mild with only two grade 3 (ECOG) toxicities and with no grade 4 toxicity observed during this study. PMID- 8229157 TI - Reversal of the influence of group Ib afferents from plantaris on activity in medial gastrocnemius muscle during locomotor activity. AB - 1. Rhythmic locomotor activity was evoked in clonidine-treated acute and chronic spinal cats, and the effect of stimulating group I afferents from the plantaris muscle on the timing and magnitude of bursts in medial gastrocnemius (MG) motoneurons was examined. 2. The locomotor rhythm was entrained when group I afferents in the plantaris nerve were electrically stimulated with trains of stimuli presented at rates above and below the intrinsic frequency of the rhythmic activity. During entrainment at rates higher than the intrinsic frequency, a burst of activity in ipsilateral MG motoneurons was initiated approximately 40 ms after the onset of each stimulus train. At lower rates of entrainment the onset of MG bursts preceded the onset of the stimulus trains, and each stimulus train had an excitatory effect on the MG burst with a latency in the range of 30-50 ms. A similar excitatory effect was observed when the stimulus trains were triggered at a preset delay after the endogenous generation of the MG bursts. 3. The excitatory action of plantaris group I afferents on the MG motoneurons was only seen during periods of locomotor activity. In the absence of rhythmic activity, the same stimulus trains reduced any ongoing tonic activity in MG motoneurons. 4. Vibration of the plantaris muscle to preferentially activate group Ia afferents neither entrained the locomotor rhythm nor increased the magnitude of the MG bursts. 5. We conclude that during locomotor activity, input from group Ib afferents of the plantaris muscle has an excitatory action on the system of interneurons generating the extensor bursts, i.e., on the extensor half center of the central rhythm generator.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229158 TI - Modeling the gastric mill central pattern generator of the lobster with a relaxation-oscillator network. AB - 1. The gastric mill central pattern generator (CPG) controls the chewing movements of teeth in the gastric mill of the lobster. This CPG has been extensively studied, but the precise mechanism underlying pattern generation is not well understood. The goal of this research was to develop a simplified model that captures the principle, biologically significant features of this CPG. We introduce a simplified neuron model that embodies approximations of well-known membrane currents, and is able to reproduce several global characteristics of gastric mill neurons. A network built with these neurons, using graded synaptic transmission and having the synaptic connections of the biological circuit, is sufficient to explain much of the network's behavior. 2. The cell model is a generalization and extension of the Van der Pol relaxation oscillator equations. It is described by two differential equations, one for current conservation and one for slow current activation. The model has a fast current that may, by adjusting one parameter, have a region of negative resistance in its current voltage (I-V) curve. It also has a slow current with a single gain parameter that can be regarded as the combination of slow inward and outward currents. 3. For suitable values of the fast current parameter and the slow current parameter, the isolated model neuron exhibits several different behaviors: plateau potentials, postinhibitory rebound, postburst hyperpolarization, and endogenous oscillations. When the slow current is separated into inward and outward fractions with separately adjustable gain parameters, the model neuron can fire tonically, be quiescent, or generate spontaneous voltage oscillations with varying amounts of depolarization or hyperpolarization. 4. The most common form of synaptic interaction in the gastric CPG is reciprocal inhibition. A pair of identical model cells, connected with reciprocal inhibition, oscillates in antiphase if either the isolated cells are endogenous oscillators, or they are quiescent without plateau potentials, or they have plateau potentials but the synaptic strengths are below a critical level. If the isolated cells have widely differing frequencies (or would have if the cells were made to oscillate by adjusting the fast currents), reciprocal inhibition entrains the cells to oscillate with the same frequency but with phases that are advanced or retarded relative to the phases seen when the cells have the same frequency. The frequency of the entrained pair of cells lies between the frequencies of the original cells. The relative phases can also be modified by using very unequal synaptic strengths.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229159 TI - Neural encoding of single-formant stimuli in the cat. I. Responses of auditory nerve fibers. AB - 1. We have studied auditory responses to a set of speech-related narrowband sounds, single-formant stimuli (SFSs), in populations of auditory nerve fibers (ANFs). An analytic method was developed to extract the envelope of temporal discharge patterns of the ANF responses to nonsinusoidally modulated stimuli, whose spectra have multiple clusters of components. Such responses are often encountered in the auditory system when complex stimuli are used and have traditionally been studied by analyzing the fundamental component of the responses. 2. The envelope modulation in the SFSs is shown to be represented by the response patterns of ANFs. When the whole ANF population is considered, the information on modulation in stimulus envelope does not disappear at the highest sound level tested at all best frequencies (BFs) we studied (1-10 kHz). The representation is the best at medium sound levels and degrades at high sound levels. Low/medium-spontaneous rate (SR) ANFs showed greater envelope modulation in their responses at high sound levels than do high-SR ANFs. The quality of the representation at high sound levels is, on average, proportional to BF threshold of an ANF. On the basis of populations of ANFs with all SRs, the envelope modulation in the SFSs is represented over a wide range of sound levels. 3. We found that low-BF ANFs differ from high-BF ANFs in representing envelope modulation in the SFSs. For ANFs with BFs less than approximately 6 kHz, information on stimulus envelope is not only contained in spectral components near direct current but also in components at the vicinities of frequencies equal to BF and its multiples. In fact, for ANFs with BFs < 3 kHz, the contribution from spectral components centered at BF to overall response modulation is greater than that from spectral components near direct current. These findings indicate that, by using measures solely based on the fundamental component, the amount of modulation in the responses to narrowband stimuli is underestimated for low-BF ANFs. 4. Off-BF stimulation of ANFs with SFSs was found to result in increased envelope modulation in responses at high sound levels. The further away the stimulus is centered relative to unit BF, the greater the modulation it induces, provided that the stimulus is capable of exciting the unit. An SFS centered as close as 15% off unit BF can produce a significant increase in the modulation of responses at very high sound levels.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229160 TI - Synaptic integration in an excitable dendritic tree. AB - 1. Compartmental modeling experiments were carried out in an anatomically characterized neocortical pyramidal cell to study the integrative behavior of a complex dendritic tree containing active membrane mechanisms. Building on a previously presented hypothesis, this work provides further support for a novel principle of dendritic information processing that could underlie a capacity for nonlinear pattern discrimination and/or sensory processing within the dendritic trees of individual nerve cells. 2. It was previously demonstrated that when excitatory synaptic input to a pyramidal cell is dominated by voltage-dependent N methyl-D-aspartate (NMDA)-type channels, the cell responds more strongly when synaptic drive is concentrated within several dendritic regions than when it is delivered diffusely across the dendritic arbor. This effect, called dendritic "cluster sensitivity," persisted under wide-ranging parameter variations and directly implicated the spatial ordering of afferent synaptic connections onto the dendritic tree as an important determinant of neuronal response selectivity. 3. In this work, the sensitivity of neocortical dendrites to spatially clustered synaptic drive has been further studied with fast sodium and slow calcium spiking mechanisms present in the dendritic membrane. Several spatial distributions of the dendritic spiking mechanisms were tested with and without NMDA synapses. Results of numerous simulations reveal that dendritic cluster sensitivity is a highly robust phenomenon in dendrites containing a sufficiency of excitatory membrane mechanisms and is only weakly dependent on their detailed spatial distribution, peak conductances, or kinetics. Factors that either work against or make irrelevant the dendritic cluster sensitivity effect include 1) very high resistance spine necks, 2) very large synaptic conductances, 3) very high baseline levels of synaptic activity, and 4) large fluctuations in level of synaptic activity on short time scales. 4. The functional significance of dendritic cluster sensitivity has been previously discussed in the context of associative learning and memory. Here it is demonstrated that the dendritic tree of a cluster-sensitive neuron implements an approximative spatial correlation, or sum of products operation, such as that which could underlie nonlinear disparity tuning in binocular visual neurons. PMID- 8229161 TI - Adaptive control for backward quadrupedal walking. III. Stumbling corrective reactions and cutaneous reflex sensitivity. AB - 1. Four cats were trained to walk backward (BWD) and forward (FWD) on a motorized treadmill. Mechanical (taps) or electrical (pulses) stimuli were applied to the dorsal or ventral aspect of the hind paw during swing or stance. Hindlimb kinematic data, obtained by digitizing 16-mm high-speed film, were synchronized with computer-analyzed electromyograms (EMG) recorded from anterior biceps femoris (ABF), vastus lateralis (VL), lateral gastrocnemius (LG), tibialis anterior (TA), and semitendinosus (ST). Responses to taps and pulses, as well as the modulation in cutaneous reflex sensitivity to pulses, were described for both walking directions and stimulus locations. 2. After dorsal taps that obstructed FWD swing, the hindlimb initially drew back away from the obstacle with knee flexion and ST activation, ankle extension with TA suppression and LG activation, and hip extension with ABF facilitation. Next, the limb was raised over the obstacle with resumed TA activity and enhanced knee and ankle flexion, and then compensatory knee and ankle extension positioned the limb for the ensuing stance phase. 3. For ventral taps that obstructed BWD swing, the initial response also tended to draw the limb away from the obstacle with hip and ankle flexion and TA facilitation and reduced knee flexion with weak VL facilitation and suppression of ST activity. Next, ST activity resumed as knee and ankle flexion raised the limb over the obstacle, and then compensatory extension completed the swing phase for BWD walking. Thus the initial kinematic and EMG responses to obstacles were opposite for BWD versus FWD swing, and these responses were consistent with active avoidance of the obstacles. Responses during BWD walking were subtle, however, compared with those for FWD. 4. After nonobstructing taps (ventral FWD, dorsal BWD), ST and TA activation and knee and ankle flexion were coincident, demonstrating that the aforementioned differences in responses to obstructing obstacles were not simply location dependent. Regardless of the direction of walking or the location of stimulation, taps applied during stance had little immediate kinematic effect, but the subsequent swing phase was usually exaggerated, as if the response was programmed to avoid any lingering obstacle. 5. Electrical pulses did not elicit the full-blown responses typically evoked by taps. The sequencing in activation of ST and TA characteristic after laps was absent after pulses, and there were rarely dramatic kinematic responses to pulses like those easily elicited by taps. There were, in fact, few differences in responses to electrical stimulation for BWD versus FWD walking.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229162 TI - Response properties of neurons in temporal cortical visual areas of infant monkeys. AB - 1. Inferior temporal cortex (IT) is a "high-order" region of primate temporal visual cortex implicated in visual pattern perception and recognition. To gain some insight into the development of this area, we compared the properties of single neurons in IT in infant monkeys ranging from 5 wk to 7 mo of age with those of neurons in IT in adult animals. Both anesthetized and awake behaving paradigms were used. 2. In immobilized infant monkeys under nitrous oxide anesthesia, the incidence of visually responsive cells was markedly less than in adult monkeys studied under similar conditions. In infants 4-7 mo of age, only half of IT neurons studied were visually responsive, compared with > 80% in adult monkeys. In monkeys < 4 mo old, even fewer (< 10%) could be visually driven. "Habituation" of IT cells to repeated stimulus presentation appeared more pronounced in infant monkeys under nitrous oxide anesthesia than in adult animals. 3. IT cells in the anesthetized infant monkeys that did respond showed receptive field properties similar to those of responsive adult IT neurons studied under similar conditions. Two thirds of the receptive fields plotted in the anesthetized 4 to 7-mo-old group were bilateral, and median field size did not differ between the infants and comparable adult groups, being approximately 20 degrees on a side in each case. 4. In contrast to the results obtained under anesthesia, most IT cells in alert infant monkeys 5 wk-7 mo of age (80%) were responsive to visual stimuli, and this incidence of visually responsive IT neurons did not differ from that obtained in awake adult macaques. However, response magnitude, measured as spikes per second above baseline rate, was significantly lower in the infant alert sample than in the adult control (5.2 vs. 12.6 spikes/s, mean +/- SE, deviation from spontaneous rate, respectively). 5. In addition to having lower magnitudes of visual response, IT cells in the awake infants also tended to have longer and more variable latencies. The overall mean for the infant cells was 196 ms, compared with an overall mean of 140 ms for IT neurons in the alert control adult. 6. Although the magnitude of response of neurons in alert infant IT cortex was lower overall, the incidence and features of stimulus selectivity shown by alert infant IT neurons were strikingly similar to those of IT cells of both anesthetized and unanesthetized adult monkeys.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229163 TI - Parallel, interdependent channels for location and orientation in sensorimotor transformations for reaching and grasping. AB - 1. Subjects were presented with a cylinder, whose orientation with respect to the vertical and location in space varied from trial to trial. They grasped a similar cylinder in their hands and were instructed to align the grasped cylinder with the target cylinder. In some experiments the task was performed from memory, and subjects attempted to reproduce both location and orientation of the cylinder. In others, they attempted to reproduce only its orientation, either from memory or while the cylinder was in view. 2. Multivariate linear regression analysis was used to determine persistent and variable errors in performance. This analysis related the subjects' performance (reproduced orientation) to target parameters (location and orientation). 3. We have interpreted the experimental results starting from the assumption that there are two parallel neural processes underlying reaching and grasping: one relating proximal arm motion to target location and the other relating distal hand motion to target orientation. 4. Variable errors did not vary with task conditions, even when subjects were asked to dissociate target orientation from target location by matching target orientation with the arm at the side, irrespective of the location of the target. This finding suggests that the neural transformations involving target location and target orientation are performed in parallel. 5. Persistent errors did vary with task condition. The subjects made the largest errors in matching target orientation when the target was in view, but they were asked to match its orientation at a location that differed from that of the target. These errors depended mostly on the elevation of the target and on its slant (inclination relative to the vertical). They were related to the posture of the arm in a manner that suggested that matching orientation is influenced by both extrinsic (spatial) and intrinsic (arm posture) parameters. 6. The fact that persistent errors depend on target location and on arm posture as well as on target orientation implies that the neural transformation from target orientation to hand orientation is not independent of the transformation dealing with target location. PMID- 8229164 TI - Modulation of monosynaptic excitation in the neonatal rat spinal cord. AB - 1. The effects of high-frequency (5-50 Hz) stimulation of dorsal root afferents on monosynaptic excitation of alpha motoneurons was studied in the in vitro spinal cord preparation of the neonatal rat, using sharp-electrode intracellular recordings. 2. Double pulse stimulation of dorsal root afferents induced severe depression of testing excitatory postsynaptic potentials (EPSPs) at each of the tested interstimulus intervals (15 ms-5 s). After perfusion of the preparation with low-calcium, high-magnesium Krebs saline, the amplitude of the conditioning EPSPs was markedly decreased and the testing EPSPs exhibited substantial facilitation that was maximal at the 20-ms interval and that was accompanied by depression at intervals > or = 60-100 ms. 3. Short-duration stimulus trains applied to dorsal root afferents normally induced tetanic depression of the intracellularly recorded monosynaptic EPSPs. Switching the bathing solution to low-calcium, high-magnesium saline decreased the control EPSP and induced facilitation and then tetanic potentiation (TP) of the EPSPs within the applied train. The magnitude of potentiation (% potentiation) of these EPSPs depended on the interpulse interval of the short stimulus train and on the degree of attenuation of the unpotentiated control EPSP after the solution was changed from normal- to low-calcium Krebs solution. 4. Long-duration stimulus trains applied to dorsal root afferents at 5-10 Hz induced marked depression of monosynaptic EPSPs during the train. The depression was alleviated after cessation of the tetanic stimulation and was followed in some cases by slight posttetanic potentiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229165 TI - Presynaptic glutamate receptors depress inhibitory postsynaptic transmission in lobster neuromuscular synapse. AB - 1. We examined the functional role of GTP-coupled glutamate receptor (GluB-R) in the presynaptic membrane of lobster neuromuscular synapse. 2. Injection of guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), a hydrolysis-resistant analogue of GTP, into the excitatory axon mimicked the presynaptic glutamate response and effectively suppressed excitatory postsynaptic potentials or excitatory postsynaptic currents (EPSCs). 3. Statistical analysis revealed that the coefficient of variation (standard deviation divided by the mean of EPSC amplitude) was increased after injection of GTP gamma S into the excitatory axon, indicating a presynaptic inhibition of transmitter release. 4. The effect of glutamate on inhibitory postsynaptic potentials (IPSPs) or inhibitory postsynaptic currents (IPSCs) was studied when the postsynaptic glutamate receptors were blocked by the Joro spider toxin (JSTX). Glutamate depressed IPSPs or IPSCs in the JSTX-treated preparation. Furthermore, repetitive stimulation of the excitatory nerve produced effective inhibition of IPSCs. 5. Quisqualate and kainate suppressed IPSCs in a similar way to glutamate. In contrast, N-methyl-D aspartate, ibotenic acid, trans-D,L-1-amino-1,3-cyclopentanedicarboxyloc acid, and 2-amino-4-phosphonobutanate had no effect on GluB-R. 6. Our results indicate that GluB-R, which exists in both excitatory and inhibitory nerve terminals, regulates transmitter release by a presynaptic inhibitory mechanism. PMID- 8229166 TI - Response characteristics of lamb pontine neurons to stimulation of the oral cavity and epiglottis with different sensory modalities. AB - 1. To better understand sensory information processing in pontine neurons that receive afferent fiber terminations from oral cavity and upper airway receptors, we investigated the response characteristics of single neurons to stimulation of the oral cavity and epiglottis with different stimulus modalities. These response characteristics were then compared with previously recorded response properties of neurons located in other brain stem regions that receive oral cavity and upper airway sensory inputs. 2. Receptive field sizes of pontine neurons were mapped, and responses to mechanical, thermal, and chemical stimuli were determined. A total of 47 neurons were isolated and most neurons were located near the dorsomedial border of the rostral trigeminal subnucleus oralis and caudal principal trigeminal nucleus. The likelihood that a particular stimulus modality would elicit a response was somewhat dependent on a neuron's location. Neurons that responded to chemical stimuli were always located outside the trigeminal nucleus, whereas neurons that responded exclusively to mechanical or thermal stimuli were more frequently located in the trigeminal nucleus. Receptive fields were mapped for 45 of the 47 neurons. Forty-three of the neurons had a single ipsilateral receptive field and > 80% of the receptive fields were > 100 mm2. The majority of neurons responded to only one of the three stimulus modalities. The remaining neurons were multimodal and the combination of stimulus modalities most frequently observed was mechanical and chemical. 3. Mechanical stimuli were the most effective of the three stimulus modalities, eliciting responses in > 65% of the neurons. Neurons that responded to mechanical stimuli were generally rapidly adapting and a moving stimulus was more effective than a punctate stimulus. Mechanosensitive neurons that also responded to chemical stimuli exhibited larger mean response frequencies than mechanosensitive neurons that did not respond to chemical stimuli. Chemical stimuli elicited responses in about half the neurons. A greater percentage of neurons with receptive fields on the epiglottis than neurons with oral cavity receptive fields responded to chemical stimuli. The effectiveness of a chemical stimulus was dependent on a neuron's receptive field. NH4Cl was the most effective stimulus for neurons with receptive fields located in the oral cavity, whereas KCl was more effective for neurons with receptive fields on the epiglottis. Thermal stimuli were relatively ineffective whatever the location of a neuron's receptive field. The majority of neurons showed an increase in response frequency to cooling the receptive field and in all thermosensitive neurons the response was restricted to the dynamic phase of thermal stimulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229167 TI - Rapid amplification and facilitation of mechanosensory discharge in Aplysia by noxious stimulation. AB - 1. Little is known about modulation of action potential discharge in Aplysia mechanosensory neurons during defensive responses. The present studies examined rapid effects of noxious stimulation (occurring within 0.5-10 s) on the number of action potentials evoked by test stimuli delivered to the tail. Responses were monitored in the somata of mechanonociceptors in the pleural ganglion. A major hypothesis to be tested was that an important function of previously described alterations of membrane conductances in the sensory neuron soma is to generate an after-discharge that amplifies sensory signals during severe noxious stimulation of the cell's receptive field. 2. Discharge of spikes evoked by a moderate tap to one part of a sensory neuron's receptive field on the tail was enhanced by strong shock delivered 10 s earlier to another part of the field. Part of this enhancement appears to be due to a decrease in conduction block in central regions of the sensory neuron. 3. Repeated delivery of innocuous, moderately intense tail shock at 5-s intervals caused a progressive increase ("wind-up") of discharge, whereas repeated delivery of weak tail shock had no significant effect on discharge. In some cases the increase in action potential number involved a buildup of afterdischarge. 4. A single strong tail pinch sometimes induced an afterdischarge lasting < or = 2 s. Afterdischarge could also be induced in the isolated nervous system by intense electrical stimulation of the nerve containing the sensory neuron's main axon. 5. Several observations suggest that afterdischarge requires cooperative effects of a relatively large number of coactivated fibers in the test pathway. In contrast to pinching stimuli (which stimulated a larger part of the tail), intense, punctate stimulation with von Frey hairs failed to produce afterdischarge. Weaker tail or nerve stimulation failed to produce afterdischarge, even when short-latency, high-frequency discharge was evoked in the sensory neuron. 6. Cooperative effects on afterdischarge may differ from those involved in activity-dependent enhancement of presynaptic facilitation because simultaneous pairing of high-frequency activation of a single test sensory neuron with strong stimulation of a peripheral nerve lacking an axon of the tested sensory neuron was not sufficient to produce afterdischarge. The cooperative effects on afterdischarge may function to encode information about both the severity and spatial extensiveness of an injury. 7. Artificial hyperpolarization of the soma often reversibly reduced or abolished afterdischarge evoked by stimulating the nerve or tail. Thus the afterdischarge is often generated in or near the sensory neuron soma.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229168 TI - Activity-dependent depression of mechanosensory discharge in Aplysia. AB - 1. Inhibition of action potential discharge in Aplysia mechanosensory neurons after noxious stimulation has not been described previously. The present studies investigated depressive effects of prolonged noxious stimulation and repetitive intracellular activation on the number and latency of action potentials evoked by test stimuli applied to the tail or the nerve innervating the tail. Action potential discharge was monitored in the somata of mechanonociceptors in the pleural ganglia. 2. Repeated brief pinches delivered at 5-s intervals to a sensory neuron's receptive field on the tail initially caused intense activation (10-25 spikes recorded in the soma) followed by a progressive decrease or "wind down" of spike number during subsequent pinches. 3. Repeated application to the tail of noxious shock that caused intense activation of sensory neurons (10-22 spikes during the initial shock) produced progressive wind-down of discharge similar to that produced by repeated tail pinch. However, sensory neurons that showed lower activation (1-9 spikes) to the same shock displayed wind-up of discharge during the 10 shocks. These results suggested that prolonged, intense activation depresses subsequent action potential discharge. 4. Changes in the time required for spikes evoked in the tail to reach the central soma were used as an indicator of changes in the excitability and/or conduction velocity of peripheral branches. Repeated pinch within a sensory neuron's receptive field caused an increase in the latency of discharge elicited by test shocks within the receptive field that lasted > or = 10 min. Repetitive intracellular stimulation of the sensory neuron soma caused a similar increase in latency. 5. Repetitive soma activation decreased the number of spikes evoked 10 s later by a test shock in the sensory neuron's receptive field, indicating that spike activity depresses the initiation and/or conduction of spikes in peripheral branches. Surprisingly, repeated pinch to the receptive field caused no significant change in the number of spikes evoked by the same test shock. This difference suggests that tail pinch produces concomitant facilitatory effects that oppose the depressive effects of intense spike activity. 6. Depressive effects of repeated pinch and repetitive soma activation were expressed in the axon between the receptive field and the CNS. Spikes evoked by brief test shocks delivered to the nerve containing the axon of the recorded sensory neuron showed a transient increase in latency (perhaps due to a decrease in conduction velocity) after either procedure. Repeated pinch, but not repetitive soma activation, also caused an increase in spike threshold in the nerve.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229169 TI - Sensitization of the gill and siphon withdrawal reflex of Aplysia: multiple sites of change in the neuronal network. AB - 1. Recent studies have emphasized the major contribution of interneuronal transmission to the mediation and learning-associated modulation of the gill and siphon withdrawal (GSW) reflex of Aplysia. We wish to provide more direct support for the hypothesis that inhibitory junctions are crucial sites of plasticity. 2. In parallel experiments we investigated modulation at five major sites of synaptic transmission in the GSW network: 1) from sensory neurons to motor neurons, 2) from sensory neurons to excitatory interneurons (INTs+) 3) from INTs+ to motor neurons (MNs), 4) from inhibitory interneurons (INTs-) to INTs+, and 5) from INTs+ to INTs-. 3. While recording simultaneously from a single sensory neuron of the LE cluster, an INT+, and a MN, we found that both LE-MN and LE INTs+ synapses were facilitated by the activation of modulator neurons by stimulation of the left pleuroabdominal connective (185 and 93%, respectively) as well as by serotonin (5-HT) (191 and 84%). Junctions of the second type were therefore less facilitated. The difference in the magnitude of facilitation at these two sites is an indication of a branch-specific, differential efficacy in the modulation of different central synapses made by a single neuron. 4. Although INT(+)-MN junctions have the capacity to display marked posttetanic potentiation, they are not significantly potentiated after connective stimulation. Sensitization of the GSW reflex is therefore not necessarily accompanied by a modification of transmission at these synapses. 5. Inhibitory transmission to INTs+ is significantly reduced by connective stimulation (36%) and by 5-HT (71%). This supports the hypothesis that a reduction of feedback inhibition into INTs+ is a major mechanism of reflex sensitization and may account for the increased evoked firing of INTs+ that is observed after connective stimulation. 6. The excitatory input to INTs- is selectively decreased by 5-HT (50%) and by the molluscan neuropeptide small cardioactive peptide B (38%). This latter effect, which could produce disinhibition of INTs+, may explain the previous observation that this peptide is able to potentiate the evoked input to MNs of the reflex at a concentration (1 microM) that fails to modify monosynaptic sensory-motor transmission. 7. These results indicate that transmission through a small neuronal network that mediates a withdrawal reflex in Aplysia may be modulated at multiple sites and by different mechanisms. These mechanisms include: 1) branch specific facilitation of sensory neuron outputs and 2) inhibition of INT(-)-INT+ inhibitory postsynaptic potentials by endogenous modulatory neurons and by 5 HT.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229170 TI - Effects of cerebral neuron C-PR on body postural muscles associated with a food induced arousal state in Aplysia. AB - 1. Firing of cerebral neuron, C-PR, produced complex bilateral movements of various regions of the body of the marine mollusc Aplysia californica. The movements were similar to those seen when the animal assumes the head-up feeding posture during food-induced arousal. Muscles of the neck largely contracted in transverse and longitudinal directions, and large transverse movements were also induced in the middle part of the foot. On the other hand, firing of C-PR appeared to relax the anterior part of the foot in transverse and longitudinal directions. 2. We identified pedal-ganglion motor neurons that innervate various regions of the animal, and explored the synaptic connections of C-PR with these neurons. Firing of C-PR produced synaptic potentials bilaterally in most of the identified motor neurons. 3. Motor neurons for the neck were largely excited by C PR firing. C-PR firing also excited the motor neurons that produce transverse movements of the middle part of the foot. On the other hand, C-PR inhibited the spontaneous spike activity of the motor neurons for the anterior part of the foot. 4. One neck motor neuron was found to receive a monosynaptic excitatory postsynaptic potential (EPSP) from C-PR, but the postsynaptic potentials (PSPs) induced by C-PR in the other identified motor neurons were mediated polysynaptically. 5. We also found that the C-PR can modulate movements evoked by firing of the motor neurons for the ipsilateral neck and anterior foot. C-PR enhanced both transverse and longitudinal contractions of the neck. 6. For the anterior foot region, C-PR had different modulatory effects on the longitudinal and the transverse contractions. C-PR largely enhanced or initially depressed and then enhanced longitudinal contractions, whereas C-PR depressed transverse contractions. 7. The overall results support the hypothesis that C-PR is involved in controlling the head-up posture when the animal is aroused by food. PMID- 8229171 TI - Membrane properties of rat subicular neurons in vitro. AB - 1. Conventional intracellular recordings were performed in rat hippocampal slices to investigate the electrophysiological properties of subicular neurons. These cells had a resting membrane potential (RMP) of -66 +/- 7.2 mV (mean +/- SD; n = 50), input resistance of 23.6 +/- 8.2 M omega (n = 51), time constant of 7.1 +/- 1.9 ms (n = 51), action potential amplitude of 85.8 +/- 13.8 mV (n = 50), and duration of 2.9 +/- 1.2 ms (n = 48). Analysis of the current-voltage relationship revealed membrane inward rectification in both depolarizing and hyperpolarizing direction. The latter type was readily abolished by Cs+ (3 mM; n = 6 cells). 2. Injection of depolarizing current pulses of threshold intensity induced in all subicular neurons (n = 51) recorded at RMP a burst of two to three fast action potentials (frequency = 212.7 +/- 90 Hz, n = 13 cells). This burst rode on a slow depolarizing envelope and was followed by an afterhyperpolarization and later by regular spiking mode once the pulse was prolonged. Similar bursts were also generated upon termination of a hyperpolarizing current pulse. 3. The slow depolarization underlying the burst resembled a low-threshold response, which in thalamic cells is caused by a Ca2+ conductance and is contributed by the Cs(+) sensitive inward rectifier. However, bursts in subicular cells persisted in medium containing the Ca(2+)-channel blockers Co2+ (2 mM) and Cd2+ (1 mM) (n = 5 cells) but disappeared during application of TTX (1 microM; n = 3 cells). Hence they were mediated by Na+. Blockade of the hyperpolarizing inward rectification by Cs+ did not prevent the rebound response (n = 3 cells). 4. Our findings demonstrate that intrinsic bursts, presumably related to a "low-threshold" Na+ conductance are present in rat subicular neurons. Similar intrinsic characteristics have been suggested to underlie the rhythmic activity described in other neuronal networks, although in most cases the low-threshold electrogenesis was caused by Ca2+. We propose that the bursting mechanism might play a role in modulating incoming signals from the classical hippocampal circuit within the limbic system. PMID- 8229172 TI - Synaptic noise and multiquantal release at dendritic synapses. AB - 1. The quantal nature of inhibitory synaptic noise recorded intracellularly from the lateral dendrite of the goldfish Mauthner cell was studied, using new detection and measurement procedures that eliminate operator intervention. In addition, we employed an analytical algorithm, not previously applied to this problem, which treats composite amplitude distributions as mixtures of gaussians of unknown separations and variances. 2. As in the soma of this neuron, the dendritic inhibitory noise is quantal, with the exception that in the dendrite multiple equally spaced classes may persist in the presence of tetrodotoxin (TTX), an observation that may be correlated with the finding that the inhibitory afferents at this level often contain more than one release site. The validity of the analysis was confirmed by superfusing with saline containing low calcium and high magnesium, which reduces composite histograms that are gaussian mixtures to a single class, equal in amplitude to that of the first component detected in the control. 3. These results suggest that spontaneous exocytotic events may be synchronized at adjacent active zones within single terminals and that lowering the probability of release by reducing calcium may then be a more effective method for isolating single miniature events than is TTX. PMID- 8229173 TI - Zinc enhances GABAergic transmission in rat neocortical neurons. AB - 1. Intracellular recordings were made in layer II-III neurons of rat neocortical slices maintained in vitro. The effect of bath application of zinc (50-300 microM) on evoked synaptic activity and passive membrane properties was examined. 2. Excitatory postsynaptic potentials (EPSPs) mediated by N-methyl-D-aspartate (NMDA) and non-NMDA receptors were recorded in response to electrical stimulation. Zinc did not affect either type of EPSP. Resting membrane potential, repetitive firing properties, and input resistance were not altered by zinc. 3. Inhibitory postsynaptic potentials (IPSPs) were enhanced after zinc application. Zinc also induced generation of large amplitude spontaneous gamma-aminobutyric acid-A (GABAA)- and GABAB-mediated IPSPs. Postsynaptic responses to iontophoretically applied GABA were unaffected. In the presence of zinc, GABAergic synaptic potentials could result in generation of action potentials. 4. Directly evoked IPSPs recorded in the presence of the excitatory amino acid receptor blockers 6-cyano-7-nitroquinoxaline-2,3-dione and 2-amino-5 phosphonovaleric acid were enhanced by zinc. Under these conditions spontaneous IPSPs with superimposed action potentials were present. Baclofen, in the presence of zinc, reduced the amplitude of evoked IPSPs. 5. These results indicate that zinc may be an endogenously occurring neuromodulator. Zinc appears to enhance GABAergic IPSPs by increasing the excitability of inhibitory interneurons, thus resulting in increased GABA release. PMID- 8229174 TI - Estimation of passive horizontal linear whole-body displacement in humans. AB - 1. Passive linear self-motion estimation along the X and Y axes was investigated in human subjects. 2. A target was viewed from a distance of 0.8 or 2.4 m from the starting point. Subjects were then blindfolded and transported toward the target on a distance of 3.2 m and back to the start. Acceleration was constant: +/- 0.2 or +/- 1 m/s2. The subjects pushed a button on both outward and return paths, when they passed the previously seen target. 3. The results showed anticipation of the target on the outward path, and not on the return. This was identical for both axes and both accelerations. 4. The data are in accord with a model of double integration of the otolith signal, suggesting that linear path integration is a basic sensory mechanism. PMID- 8229175 TI - Regular oscillations of synaptic activity in spinal networks in vitro. AB - 1. Spontaneous synaptic potentials were recorded in motoneurons grown in organotypic slice cultures of embryonic rat spinal cord. In 71 of 85 cells these potentials appeared without obvious temporal structure (random patterns); in the remaining 14 cells they appeared in bursts (rhythmic patterns). 2. Random activity patterns could be converted into rhythmic patterns by treating the cultures with strychnine, bicuculline, or both. The excitatory amino acid N methyl-D-aspartate (NMDA) transiently increased the rate of spontaneous synaptic activity without inducing rhythmic patterns. The NMDA antagonist 7-chloro kynurenate reduced the burst rate while leaving the burst length unchanged in rhythmic patterns. In random patterns it reduced the rate of spontaneous synaptic activity by 68%. 3. Histograms of interevent times of the random patterns were best fitted by the sum of two expontentials, suggesting that the random type of activity could not be described simply as a Poisson process but involved at least one additional mechanism. 4. Rhythmic patterns consisted of bursts of activity with a mean burst length of 2.2 s that were separated by interburst intervals with a mean length of 6.6 s. Within the bursts autocorrelograms revealed regular oscillations with a mean period of 226 ms in 6 of 11 experiments with rhythmic patterns. The period showed little variation between individual experiments (202 288 ms). In random patterns no oscillations were detected. 5. Within the spontaneous bursts the excitatory postsynaptic potentials (EPSPs) progressively declined in amplitude. A corresponding depression of EPSPs was observed when trains of electrical stimuli were applied at 5 Hz to the dorsal horns of the spinal cord slices.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229176 TI - Taste responses of neurons in the nucleus of the solitary tract of awake rats: an extended stimulus array. AB - 1. Fifty-seven taste neurons were isolated in the nucleus solitary tract (NST) and tested with 15 sapid chemicals. On average, NST neurons responded well to NaCl, sucrose, monosodium L-glutamate (MSG), NaNO3, and glycine (mean = 8.2-11.0 spikes/s). Mean responses to KCl, NH4Cl, HCl, malic acid, and quinine HCl (QHCl) were low (mean = 0.7-2.9). The average responses to the other stimuli (citric acid, MgCl2, fructose, maltose, and polycose) fell between these extremes (mean = 4.3-5.1). 2. On the basis of the largest response to the four standard stimuli, the neurons were classified as follows: 15 NaCl-best, 23 sucrose-best, 17 citric acid-best, and 2 QHCl-best. 3. The NaCl-best neurons responded robustly and nearly equally to the three sodium salts (mean = 15.7-20.8) but much less so and more variably to the nonsodium, chloride salts (mean = -0.1-4.6). Sucrose-best neurons responded strongly to sucrose, glycine, and MSG (mean = 13.7-17.8), but only moderately to the other sugars (fructose and maltose) and to polycose (mean = 8.4, 9.8, and 8.8, respectively). 4. Citric acid-best neurons responded moderately to citric and malic acid (mean = 9.4 and 4.7), but less so to HCl (mean = 3.1). The two QHCl-best neurons responded moderately to QHCl and MgCl2 (mean = 12.0 and 9.5), but weakly or not at all to the other stimuli (mean = -1.1 3.1). 5. Unlike parabrachial taste neurons, none of the medullary taste cells responded specifically to Cl(-)-containing chemicals. The responses that did occur to nonsodium salts were weak and variable and often occurred in either citric acid-best or QHCl-best neurons, rather than in those that responded vigorously to sodium salts. Similar relationships have been observed in anesthetized preparations. 6. A hierarchical cluster analysis for 57 neurons across 15 stimuli produced four second-order clusters that consisted primarily of NaCl-best, sucrose-best, citric acid-best, and QHCl-best neurons, respectively. Although the analysis for neurons produced only four such clusters, a similar analysis for the 15 stimuli separated the sodium salts (NaCl and NaNO3), nonsodium salts (KCL, NH4Cl, and MGCl2, sweeteners (sucrose, maltose, fructose, and glycine), acids (citric acid and malic acid), and QHCl. 7. Monosodium glutamate activated both NaCl-best and sucrose-best neurons, but the stimulus analysis clumped it with the sodium salts.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229177 TI - Thalamocortical interactions in the somatosensory system: interpretations of latency and cross-correlation analyses. AB - 1. Isolated extracellular neuronal responses to cutaneous stimulation were simultaneously recorded from corresponding peripheral representations in the ventrobasal nucleus and primary somatosensory cortex of intact, halothane anesthetized rats. Thalamic and cortical neurons representing hairy skin on the forelimb were activated by hair movements produced by a series of 50 or 100 discrete air jets. A corresponding set of neurons representing the glabrous pads of the hind paw were activated by a similar number of punctate mechanical displacements. 2. Cortical electrode penetrations were histologically reconstructed, and 118 neurons in the glabrous skin representation exhibited cutaneous responses that were categorized into supragranular, granular, or infragranular groups according to their laminar position. Minimum latencies of cortical neurons responding to glabrous skin displacement were analyzed, and significant differences were found in the distribution of minimum latencies for the different cortical layers. Mean values for minimum latencies in the infragranular and granular layers were 15.8 and 16.3 ms, respectively, whereas supragranular neurons were characterized by minimum latencies having a mean of 20 ms. The differences between these groups suggests that stimulus-induced afferent activity reaches infragranular and granular layers before contacting supragranular neurons. Average latencies were also calculated on responses occurring during the 1st 20 trials, but the cortical distributions of these values overlapped considerably, and differences between the laminar groups were not statistically significant. 3. In several recording sites, two cortical neurons were recorded simultaneously, and the response latencies of these matched pairs were often substantially different despite the similarity in laminar position. This result indicates that laminar location is not the only determinant of response latency and that serially organized circuits are distributed within, as well as between, cortical layers. 4. From a sample of 302 neurons exhibiting cutaneous responses within histologically identified regions of thalamus or cortex, a set of 143 pairs of neurons recorded simultaneously from both regions was available for cross-correlation analysis. Significant thalamocortical interactions were found in 38 neurons pairs. Analysis of these significant interactions revealed that thalamocortical connection strength, as measured by neuronal efficacy, was two to four times larger for neuron pairs having the cortical cell in granular layer IV than for neuron pairs having an extragranular layer cortical neuron. There was no difference in thalamocortical connection strength between neuron pairs containing supra- or infragranular cortical neurons. 5. Summed peristimulus time histograms revealed stimulus-locked inhibition of spontaneous activity in 4% (8/195) or cortical and 18% (20/107) of thalamic neurons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229178 TI - Focal attention produces spatially selective processing in visual cortical areas V1, V2, and V4 in the presence of competing stimuli. AB - 1. The activity of single neurons was recorded in Macaca mulatta monkeys while they performed tasks requiring them to select a cued stimulus from an array of three to eight stimuli and report the orientation of that stimulus. Stimuli were presented in a circular array centered on the fixation target and scaled to place a single stimulus element within the receptive field of the neuron under study. The timing of the cuing event permitted the directing of visual attention to the spatial location of the correct stimulus before its presentation. 2. The effects of focal attention were examined in cortical visual areas V1, V2, and V4, where a total of 672 neurons were isolated with complete studies obtained for 94 V1, 74 V2, and 74 V4 neurons with receptive-field center eccentricities in the range 1.8 8 degrees. Under certain conditions, directed focal attention results in changes in the response of V1, V2, and V4 neurons to otherwise identical stimuli at spatially specific locations. 3. More than one-third of the neurons in each area displayed differential sensitivity when attention was directed toward versus away from the spatial location of the receptive field just before and during stimulus presentation. Both relative increases and decreases in neural activity were observed in association with attention directed at receptive-field stimuli. 4. The presence of multiple competing stimuli in the visual field was a major factor determining the presence or absence of differential sensitivity. About two-thirds of the neurons that were differentially sensitive to the attending condition in the presence of competing stimuli were not differentially sensitive when single stimuli were presented in control studies. For V1 and V2 neurons the presence of only a few (3-4) competing stimuli was sufficient for a majority of the neurons studied; a majority of the V4 neurons required six to eight stimuli in the array before significant differences between attending conditions occurred. 5. For V1 and V2 neurons the neuronal sensitivity differences between attending conditions were observed primarily at or near the peak of the orientation tuning sensitivity for each neuron; the differences were evident over a broader range of orientations in V4 neurons. 6. In conclusion, neural correlates of focal attentive processes can be observed in visual cortical processing in areas V1 and V2 as well as area V4 under conditions that require stimulus feature analysis and selective spatial processing within a field of competing stimuli.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229179 TI - Respiratory modulation of the activity in postganglionic neurons supplying skeletal muscle and skin of the rat hindlimb. AB - 1. The respiratory modulation of activity in postganglionic neurons of the lumbar sympathetic outflow to skeletal muscle and hairy skin of the hindlimb was studied in anesthetized rats, either breathing spontaneously or paralyzed and artificially ventilated, using single- and multifiber recordings. The activity of the neurons was analyzed with respect to the phrenic nerve discharges or with respect to the cycle of artificial ventilation under various experimental conditions. 2. In total, 19 single and 62 multiunit preparations supplying skeletal muscle and 42 single and 95 multiunit preparations supplying skin were analyzed. Qualitatively, both populations and single- and multiunit preparations exhibited similar patterns. The majority (187/218 preparations) exhibited a depression of activity during inspiration and a peak of activity during expiration that was mostly accentuated during early expiration. The remainder exhibited a peak of activity at the transition between expiration and inspiration (n = 15) or showed no respiratory modulation (n = 16). 3. Respiratory modulation in the postganglionic neurons was similar in animals breathing spontaneously and in those that were paralyzed and artificially ventilated. 4. Systemic hypercapnia and asphyxia in most cases enhanced both the peak of activity during early expiration and the depression of activity during inspiration. 5. No peripheral (reflex) component mediated by arterial baroreceptors was found in vagotomized animals that was related to the cycle of artificial ventilation using a tidal volume of 1-2 ml and a ventilation frequency of 70 +/- 7 (SD) strokes, min-1. However, a small ventilation-related rhythm of arterial baroreceptor activity recorded from the aortic nerve was found under these conditions. 6. Respiratory modulation of postganglionic activity was similar before and after bilateral vagotomy. 7. It is concluded that respiratory modulation of sympathetic activity in rats is less differentiated than in cats. The observed differences between neurons supplying different targets are only quantitative. Possible mechanisms involved in the generation of respiratory modulation are discussed. PMID- 8229180 TI - Responses of caudal medullary raphe neurons to natural vestibular stimulation. AB - 1. Over two thirds of caudal medullary raphespinal neurons respond to electrical stimulation of the vestibular nerve, and it has been suggested that these neurons may participate in the generation of vestibulospinal and vestibulosympathetic reflexes. The objective of the present study was to determine which vestibular endorgans (semicircular canals or otolith organs) provide inputs to these cells. 2. Experiments were conducted on decerebrate cats that were baroreceptor denervated and vagotomized, and that had a cervical spinal cord transection so that inputs from tilt-sensitive receptors outside of the labyrinth did not influence the units we recorded. 3. In most experiments, vertical vestibular stimulation was used to stimulate the anterior and posterior semicircular canals and the otolith organs. The plane of whole body rotation that produced maximal modulation of a neuron's firing rate (response vector orientation) was measured at one or more frequencies between 0.1 and 0.5 Hz. Neuron dynamics were then studied with sinusoidal (0.02-1 Hz) stimuli aligned with this orientation. Alternatively, in two animals horizontal rotations at 0.5 and 1.0 Hz were employed to stimulate the horizontal semicircular canals. 4. The properties of raphespinal neurons were similar to those of a larger sample of raphe neurons studied that either could not be antidromically activated from the cervical spinal cord or were not tested for a spinal projection. In response to vertical vestibular stimulation, > 85% of caudal medullary raphe neurons had response gains that remained relatively constant across stimulus frequencies, like regularly firing otolith afferents.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229181 TI - Muscle synergies and isometric torque production: influence of supination and pronation level on elbow flexion. AB - 1. Twenty normal subjects performed a series of isometric elbow flexion (F) maximum voluntary contractions (MVC) while simultaneously maintaining one of seven targeted torque levels in the supination/pronation (S/P) degree of freedom (df). Experimental measures were torque in both df s and surface electromyograms (EMG) from brachioradialis (BRAD), triceps brachii (TB), biceps brachii (BB) short head (BBSH), and a medial and lateral site on biceps brachii long head (MED BB and LAT BB). Task effects were tested for significance using analysis of covariance models for the torque and EMG variables. Polynomial multiple regression models were developed for significant effects. The synergism among muscles was examined by statistically testing the EMG data for differing responses to the S/P torque changes across the five electrode sites. 2. The magnitude of the S/P target torque had a statistically significant effect on flexion MVC (F MVC) torque. Changes in S/P torque markedly influenced the F MVC torque magnitude, with as much as a 25% F torque decrement relative to an F MVC with an S/P torque target of 0. This suggests that the second df task affects some aspect of joint function that causes the CNS to reduce F torque capacity. 3. The S/P torque had a significant effect on EMG amplitude at all electrode sites other than TB. The EMG amplitude at the BB sites responded strongly to both F and S/P torque changes. The F+S tasks tended to facilitate BB EMG, whereas the F+P tasks tended to diminish it. The BRAD EMG, although primarily related to F torque amplitude, also was influenced by the S/P torque changes. The trends for BRAD EMG were opposite those for the BB in that BRAD EMG tended to be enhanced by the F+P tasks and reduced by the F+S tasks. 4. The synergistic pattern of stimulation (i.e., the relationship among the 5 EMG amplitude measures) was also significantly influenced by the S/P df task. Significant differences in the EMG behavior between BBSH and LAT BB were detected; however, no statistically significant differences were found between LAT BB and MED BB. The EMG behaviors at the BRAD and TB electrode sites were significantly different from those at the BB sites. 5. These inversely related responses from the BB and BRAD stress the importance of understanding the relationship between muscular activity and the function of muscles in more than one df. This finding further suggests that the synergistic relationships between muscles are dynamically related to task in all applicable dfs. It is suggested that this dynamic synergism is a natural consequence of the redundant musculoskeletal system.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229182 TI - Axon terminal hyperexcitability associated with epileptogenesis in vitro. I. Origin of ectopic spikes. AB - 1. Intracellular and extracellular recording techniques were used to study the increase in ectopic (i.e., nonsomatic) action-potential generation occurring among CA3 pyramidal cells during the kindling-like induction of electrographic seizures (EGSs) in this subpopulation of the hippocampal slice. Kindling-like stimulus trains (60 Hz, 2 s) were delivered to s. radiatum of CA3 at 10-min intervals. As EGSs developed, the frequency of ectopic firing increased markedly (by 10.33 +/- 3.29 spikes/min, mean +/- SE, P << 0.01). Several methods were applied to determine the initiation site for these action potentials within the cell (axons vs. dendrites). 2. Collision tests were conducted between known antidromic and orthodromic action potentials in CA3 cells to determine the critical period, c, for collision. Attempts were then made to collide ectopic spikes with known antidromic action potentials. At intervals less than c, ectopic spikes failed to collide with antidromic ones, in 5 of 10 cases. In these cells, this clearly indicates that the ectopic spikes were themselves of axonal origin. In the remaining five cases, ectopic spikes collided with antidromic action potentials at intervals approximately equal to c, most likely because of interactions within the complex system of recurrent axon collaterals in CA3. 3. Action potentials of CA3 pyramidal cells were simulated with the use of a compartmental computer model, NEURON. These simulations were based on prior models of CA3 pyramidal neurons and of the motoneuron action potential. Simulated action potentials generated in axonal compartments possessed a prominent inflection on their rising phase (IS-SD break), which was difficult to appreciate in those spikes generated in somatic or dendritic compartments. 4. An analysis of action potentials recorded experimentally from CA3 pyramidal cells also showed that antidromic spikes possess a prominent IS-SD break that is not present in orthodromic spikes. In addition to identified antidromic action potentials, ectopic spikes also possess such an inflection. Together with the predictions of computer simulations, this analysis also indicates that ectopic spikes originate in the axons of CA3 cells. 5. Tetrodotoxin (TTX, 50 microM) was locally applied by pressure injection while monitoring ectopic spike activity. Localized application of TTX to regions of the slice that could include the axons but not the dendrites of recorded cells abolished or markedly reduced the frequency of ectopic spikes (n = 5), further confirming the hypothesis that these action potentials arise from CA3 axons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229183 TI - The effect of bilateral cold block of the primate face primary somatosensory cortex on the performance of trained tongue-protrusion task and biting tasks. AB - 1. Studies using ablation, intracortical microstimulation (ICMS) and surface stimulation, and single-neuron recordings have suggested that the primate primary somatosensory cortex (SI) may play an important role in movement control. Our aim was to determine whether bilateral inactivation of face SI would indeed interfere with the control of tongue or jaw-closing movements. 2. Effects of reversible inactivation by cooling of face SI was investigated in two monkeys trained to perform both a tongue-protrusion task and a biting task. The cooling experiments were carried out after the orofacial representation within SI was identified by systematically defining the mechanoreceptive field of single neurons recorded in face SI. The deficits in the tongue or jaw-closing movement were evaluated by the success rates for the monkeys' performance of both tasks and by the force and electromyographic (EMG) activity recorded from the masseter, genioglossus, and digastric muscles associated with the tasks. 3. During bilateral cooling of face SI, there was a statistically significant reduction in the success rates for the performance of the tongue-protrusion task in comparison with control series of trials while the thermodes used to cool face SI were kept at 37 degrees C. Detailed analyses of force and EMG activity showed that the principal deficit was the inability of the monkeys to maintain a steady tongue-protrusive force in the force holding period during each trial and to exert a consistent tongue protrusion force between different trials. The task performance returned to control protocol levels at 4 min after commencement of rewarming. 4. Identical cooling conditions did not significantly affect the success rates for the performance of the biting task. Although the extent of the deficit was not severe enough to cause a significant reduction in successful rates for the biting task, cooling did significantly affect the ability of the monkeys to maintain a steady force in the holding period during each trial and to exert a consistent force between different trials. In one monkey the success rate of the biting task was also not affected by bilaterally cooling of face SI with a pair of larger thermodes placed on the dura over most of the face SI, face primary motor cortex (face MI), and adjacent cortical regions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229184 TI - Mechanical properties of cat soleus muscle elicited by sequential ramp stretches: implications for control of muscle. AB - 1. Force changes in areflexive cat soleus muscle in decerebrate cats were recorded in response to two sequential constant velocity (ramp) stretches, separated by a variable time interval during which the length was held constant. Initial (i.e., prestretch) background force was generated by activating the crossed-extension reflex, and stretch reflexes were eliminated by section of ipsilateral dorsal roots. 2. For the initial 400-900 microns of the first stretch, the muscle exhibited high stiffness, classically termed "short-range stiffness." This high stiffness region was followed by an abrupt reduction in stiffness, called muscle "yield," after which force remained at a relatively constant level, achieving a plateau in force. This plateau force level depended largely on stretch velocity, but this dependence was much less than proportional to the increase in stretch velocity, in that a 10-fold increase in velocity produced < 2-fold increase in plateau force. 3. In experiments where the velocities of the two sequential ramp stretches were identical, the force plateau level was the same for each stretch, regardless of the time elapsed before the second stretch (varied from 0 to 500 ms). In contrast, measures of stiffness during the initial portion of the second stretch showed time-dependent magnitude reductions. However, stiffness recovered quickly after the first stretch was completed, returning to control values within 30-40 ms. 4. In one preparation, in which the velocities of the two sequential ramp stretches were different, the force plateau elicited during the second stretch exhibited velocity dependence comparable with that recorded in the earlier single velocity studies. Furthermore, muscle yield was still evident in the case where the force change was due solely to the change in velocity and where short-range stiffness had not yet recovered fully from the initial stretch. On the basis of these findings, we argue that the classical descriptions of short-range stiffness and yield are inadequate and that the change in force that has typically been called the muscle yield reflects a transition between short-range, transient elastic behavior to steady-state, essentially viscous behavior. 5. To examine changes in the muscle's mechanical stiffness during single ramp stretches, a single pulse perturbation was superimposed at various times before, during, and subsequent to the constant velocity stretch. The force increment elicited in response to each pulse decreased relative to the initial isometric value, remained essentially constant until the end of the ramp, and then returned to its prestretch magnitude shortly (30-40 ms) after stretch termination.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229185 TI - A look into the cockpit of the fly: visual orientation, algorithms, and identified neurons. PMID- 8229186 TI - Experience-dependent plasticity in the inferior colliculus: a site for visual calibration of the neural representation of auditory space in the barn owl. AB - The optic tectum (homolog of the superior colliculus) contains mutually aligned neural maps of auditory and visual space. During development, the organization of the auditory map is guided by spatial information provided by vision: barn owls raised wearing prismatic spectacles, which optically shift the visual field and the visual map in the optic tectum, develop an auditory map that is shifted by an approximately equivalent amount, such that alignment between the two maps is preserved (Knudsen and Brainard, 1991). In this study we investigated whether this shift in the auditory map is intrinsic to the optic tectum or whether it reflects plasticity at an earlier stage in the auditory pathway. Owls were raised wearing prismatic spectacles that displaced the visual field by 23 degrees to the left or right. This manipulation alters the normal correspondence between locations in the visual field and interaural time difference (ITD), the primary cue for the azimuth of a sound source. In normal owls and in owls with at least 150 d of prism experience, extracellular unit recordings were used to assess the representations of ITD at anatomically and physiologically defined sites in the optic tectum and in the two prior stages of the auditory pathway, the external and central nuclei of the inferior colliculus (ICx and ICc). In the optic tectum of normal owls, the values of ITD to which units responded most strongly (best ITDs) varied systematically with the azimuths of unit visual receptive fields (VRFs). In the prism-reared owls, best ITDs were shifted from normal toward the values of ITD produced by sounds at the locations of the units' optically displaced VRFs. In the ICx of prism-reared owls, the representation of ITD also was shifted from normal, by an amount and in a direction that could completely account for the shift in ITD measured in the optic tectum. At some sites in the ICx, the shift in ITD tuning was apparent within the first 7-8 msec of the response; shifted tuning at such short latencies argues that the altered representation of ITD in the ICx reflects plasticity in the ascending auditory pathway, and is not the result of descending activity from higher auditory centers. In the ICc, which immediately precedes the ICx in the ascending pathway, the representation of ITD was normal. The results indicate that the visual instruction of auditory spatial tuning of neurons in the optic tectum reflects plasticity at the level of the ICx, the site where the auditory map of space is first synthesized. PMID- 8229187 TI - Stepwise repolarization from Ca2+ plateaus in neocortical pyramidal cells: evidence for nonhomogeneous distribution of HVA Ca2+ channels in dendrites. AB - Although cortical dendrites have classically been thought of as passive structures, recent evidence suggests that active conductances, including Ca2+ conductance, are also present in the dendritic membrane. To investigate this, we have recorded intracellularly in slices of rat neocortex bathed in 24 mM tetraethylammonium chloride and 1 microM TTX. Under these conditions, pyramidal neurons generated prolonged Ca2+ spikes. In computer simulations, the breakpoint voltage from which the plateau level began to repolarize was closely related to a specific region on the voltage/activation curve of the high-voltage-activated Ca2+ conductance underlying the spike. This modeling result was supported by the experimental observation that substituting Ba2+ for Ca2+ caused a hyperpolarizing shift in breakpoint voltage by 8-10 mV. Often there was stepwise repolarization from the Ca2+ spike to one or more additional plateau levels. In compartmental computer models, this could be simulated by two different mechanisms: (1) the presence of multiple, electrotonically separated sites of Ca2+ spike electrogenesis in the dendritic tree, and (2) the presence of Ca2+ channels with different voltage dependencies in the same compartment. In experiments, brief hyperpolarizing pulses could cut short the high-amplitude plateau without terminating the smaller "steps." This result could be simulated by both computer models. However, only the multicompartmental model could simulate effects of prolonged depolarizing and hyperpolarizing currents on the breakpoint. Thus, the more depolarized the breakpoint, and hence the closer the spike initiation zone to the recording site, the less it was affected by the injected current. In experiments, the ratio of the breakpoint voltages for the different plateau levels was equal to the ratio of the highest repolarization rates. These data indicate that the breakpoint voltage and the time course of repolarization were the same at all the sites of Ca2+ electrogenesis. Our findings provide strong evidence that Ca2+ spike initiation occurs at electrotonically separated "hot spots" in the dendrites, and that voltage dependence of the Ca2+ channels that underlie the spikes is the same at all sites. PMID- 8229188 TI - Developmental analysis of hippocampal mossy fiber outgrowth in a mutant mouse with inherited spike-wave seizures. AB - Neural firing patterns are an essential determinant of normal axon terminal growth and synaptic connectivity in developing afferent pathways, but the trophic role of synchronous activity in associative neural networks is less well defined. We examined the ontogeny of inherited synchronous hippocampal network discharges and mossy fiber innervation patterns at sequential stages of development in the stargazer (stg) mutant, a single-locus mouse mutation expressing generalized spike-wave epilepsy. Brief bursts of 6/sec repetitive discharges arise spontaneously on postnatal days 17-18 and persistently activate neocortical and hippocampal networks throughout adulthood. We found a striking pattern of mossy fiber recurrent axon collateral sprouting in the inner molecular layer of dentate gyrus in the adult stg hippocampus. Sprouting is not apparent until 4-6 weeks following seizure onset, but then steadily intensifies with continued synchronous activation. In the adult mutant, axon outgrowth is accompanied by a mild selective loss of hilar interneurons without gliosis. These data indicate that hypersynchronous stimulation during late postnatal brain development is linked, following a prolonged latent period, to significant fiber outgrowth and synaptic reorganization within the hippocampal formation. Since the pattern of synchronous activation in the stg mutant strongly resembles that seen in human spike-wave absence epilepsy, the synaptic plasticity described in this model has important implications for normal brain development in this common disorder. PMID- 8229189 TI - Three-dimensional visualization of the smooth endoplasmic reticulum in Purkinje cell dendrites. AB - The three-dimensional organization of the smooth endoplasmic reticulum (SER) in Purkinje cell dendrites in the chick cerebellum was investigated to assess the connectivity between its various components. Three-dimensional reconstructions of the SER within portions of Purkinje cell dendrites were performed from serial sections through the cerebellar molecular layer. In addition, semithick and thick sections (0.25-1 micron) of chick cerebellum in which the endomembrane system had been selectively stained were examined with an intermediate high-voltage electron microscope. Three-dimensional information was extracted from thick sections using stereo pairs and tomographic reconstructions from single axis tilt series. In contrast to its discontinuous appearance in thin sections, in both the three dimensional reconstructions and selectively stained thick sections the endoplasmic reticulum formed a highly interconnected network of tubules and cisterns extending throughout the dendritic shaft and into the spines. Several distinct morphological domains of endoplasmic reticulum were noted, including the hypolemmal cisternae, the endomembranes associated with the dendritic spines, and the tubular and cisternal endoplasmic reticulum in the dendritic shaft. In older chicks (aged 2 weeks), stacks of endoplasmic reticulum were also noted within the dendritic shaft. All elements of the SER within the dendritic shaft and spine appeared to be continuous with one another. The results of this study strongly suggest that the endoplasmic reticulum forms a complicated network that may be part of a single endomembrane system within Purkinje cell dendrite. PMID- 8229190 TI - Optokinetically evoked expression of corticotropin-releasing factor in inferior olivary neurons of rabbits. AB - We investigated the influence of both binocular and monocular optokinetic stimulation on the expression of corticotropin-releasing factor (CRF), a neuropeptide, in inferior olivary neurons. Rabbits were placed at the center of a cylindrical optokinetic drum that rotated at a constant velocity of 5 degrees/sec, stimulating one eye in the posterior-->anterior direction and the other eye in the anterior-->posterior direction. After 48 hr of optokinetic stimulation, rabbits were killed and the brains were prepared for immunohistochemistry. An antiserum to rat/human CRF was used to label brain-stem sections of optokinetically stimulated rabbits. The density of CRF-specific reaction product within individual olivary neurons was analyzed with a computer based densitometer. Binocular optokinetic stimulation for 48 hr caused a 20-70% increase in the optical density of "stimulated" dorsal cap neurons, measured relative to the optical density of control ("unstimulated") olivary neurons in the same histological section. This elevated expression of CRF lasted approximately 48 hr. When a delay of 18 hr was interposed between optokinetic stimulation and death of the rabbit, during which the rabbits were deprived of pattern vision, then both dorsal caps were densely labeled. Monocular optokinetic stimulation for 48 hr in the anterior-->posterior direction caused no change in CRF expression in either dorsal cap if the rabbit was immediately killed. However, following monocular anterior-->posterior stimulation, the dorsal cap contralateral to the "null-stimulated" eye was densely labeled if the rabbit was deprived of pattern vision for 18 hr prior to death. We infer that stopping prolonged "null stimulation" causes a rebound in activity in the stimulated dorsal cap. This "rebound" must have its origin at the level of the dorsal cap or more peripherally in the optokinetic pathway. PMID- 8229191 TI - Postsynaptic potentials mediated by GABA and dopamine evoked in stellate glial cells of the pituitary pars intermedia. AB - Studies have shown that many glial cells in the CNS possess receptors for neurotransmitters and that synapse-like contacts exist between glial cells and axonal terminals. Although synapse-like contacts are present between the glial cells (stellate cells) of the pituitary pars intermedia and the axons from the arcuate nucleus, it is not known whether these cells are under synaptic control. The objective of the present study was to determine whether transmitter-mediated postsynaptic potentials occurred in the stellate cells of the rat pituitary pars intermedia. Whole pituitaries were maintained in vitro, and a stimulating electrode was placed on the stalk to activate afferent fibers. Intracellular recordings were obtained with sharp microelectrodes. Stellate cells showed electrophysiological characteristics of macroglia including a resting potential more negative than -65 mV, low input resistance (< 50 M omega), and no detectable voltage-activated conductances. Single-pulse afferent nerve (stalk) stimulation evoked a [Ca2+]o-dependent postsynaptic response in the stellate cells consisting of a depolarization (< 500 msec) and a long-lasting hyperpolarization (45-75 sec). The depolarization was mimicked by GABA application and blocked by the GABAA antagonist bicuculline (100 microM). Repetitive stimulation of the stalk increased the amplitude and prolonged the GABA-mediated depolarization, during which a decrease in input resistance was observed. The hyperpolarization was mimicked by dopamine and blocked by the D2 antagonists sulpiride (2 microM) and domperidone (10 microM). Nipecotic acid (100 microM; an inhibitor of GABA uptake) or GBR 12909 (15 microM; an inhibitor of dopamine uptake) had minimal effects on the synaptic responses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229192 TI - Modulation of different K+ currents in Drosophila: a hypothetical role for the Eag subunit in multimeric K+ channels. AB - We examined the role of the ether a go-go (eag) gene in modulation of K+ currents and the possibility of its protein product Eag as a subunit in the heteromultimeric assembly of K+ channels by voltage-clamp analysis of larval muscle membrane currents. Previous DNA sequence studies indicate that the eag gene codes for a polypeptide homologous to, but distinct from, the Shaker (Sh) K+ channel subunits (Warmke et al., 1991), and electrophysiological recordings revealed allele-specific effects of eag on four identified K+ currents in Drosophila larval muscles (Zhong and Wu, 1991). Further studies of eag alleles indicated that none of the eag mutations, including alleles producing truncated mRNA messages, eliminate any of the four K+ currents, and that the mutational effects exhibit strong temperature dependence. We found that both W7, an antagonist of Ca2+/calmodulin, and cGMP analogs modulated K+ currents and that their actions were altered or even abolished by eag mutations. These results suggest a role of eag in modulation of K+ currents that may subserve integration of signals at a converging site of the two independent modulatory pathways. The Sh locus is known to encode certain subunits of the IA channel in larval muscle. The existence of multiple eag and Sh alleles enabled an independent test of the idea of Eag as a K+ channel subunit by studying IA in different double-mutant combinations. An array of allele-specific interaction between eag and Sh was observed, which reflects a close association between the Sh and eag subunits within the IA channel. Taken together, our data strengthen the possibility that the eag locus provides a subunit common to different K+ channels. The role of the eag subunit for modulating channels, as opposed to that of Sh subunits required for gating, selectivity, and conductance of the channel, suggest a combinatorial genetic framework for generating diversified K+ channels. PMID- 8229193 TI - A neurobiological model of visual attention and invariant pattern recognition based on dynamic routing of information. AB - We present a biologically plausible model of an attentional mechanism for forming position- and scale-invariant representations of objects in the visual world. The model relies on a set of control neurons to dynamically modify the synaptic strengths of intracortical connections so that information from a windowed region of primary visual cortex (V1) is selectively routed to higher cortical areas. Local spatial relationships (i.e., topography) within the attentional window are preserved as information is routed through the cortex. This enables attended objects to be represented in higher cortical areas within an object-centered reference frame that is position and scale invariant. We hypothesize that the pulvinar may provide the control signals for routing information through the cortex. The dynamics of the control neurons are governed by simple differential equations that could be realized by neurobiologically plausible circuits. In preattentive mode, the control neurons receive their input from a low-level "saliency map" representing potentially interesting regions of a scene. During the pattern recognition phase, control neurons are driven by the interaction between top-down (memory) and bottom-up (retinal input) sources. The model respects key neurophysiological, neuroanatomical, and psychophysical data relating to attention, and it makes a variety of experimentally testable predictions. PMID- 8229194 TI - Ciliary neurotrophic factor maintains motoneurons and their target muscles in developing rats. AB - Ciliary neutrophic factor (CNTF) can enhance motoneuron survival during naturally occurring cell death in the chick (Oppenheim et al, 1991). Because receptors for CNTF are expressed in both motoneurons and their target muscles (Davis et al., 1991; lp et al., 1993), both tissues are potential sites of CNTF action in development. We examined the ability of CNTF to prevent the degeneration of a neuromuscular system in developing female rats. The death of motoneurons in the spinal nucleus of the bulbocavernosus (SNB) extends postnatally and is sexually dimorphic, with many more motoneurons dying in females than in males. The bulbocavernosus (BC), a target muscle of SNB motoneurons, also degenerates postnatally in females. Female rats treated with daily injections of 1 microgram CNTF from embryonic day 22 through postnatal day 3 (P3) had 70% more SNB motoneurons on P4 than did control animals, and the number of pyknotic profiles in the SNB area was markedly reduced by CNTF. In addition, the degeneration of the BC was completely prevented by CNTF treatment of perinatal female rats. These results demonstrate that CNTF can preserve mammalian motoneurons from developmental death, but also suggest that the sparing effect of CNTF on motoneurons in vivo may be a secondary consequence of effects on target muscles. PMID- 8229195 TI - Omega-conotoxin prevents myelin-evoked growth cone collapse in neonatal rat locus coeruleus neurons in vitro. AB - The response of neonatal rat locus coeruleus neurons to contact with myelin extracts prepared from the CNS and PNS was examined. The growth cones of these neurons collapsed following contact with central myelin, but continued to elongate on contact with peripheral myelin. Central myelin elicited an increase in the intracellular free calcium concentration in these growth cones, while peripheral myelin did not. This increase appeared to require transmembrane calcium flux, since it was blocked by extracellular EGTA, and also by extracellular cobalt. These neurons express N- and L-type calcium channels. Exposure to 5 microM omega-conotoxin GVIA, a specific blocker of N-type channels, prevented both the myelin-evoked increase in growth cone calcium concentration and the collapse of growth cones. PMID- 8229196 TI - Development of neural selectivity for birdsong during vocal learning. AB - Juvenile white-crowned sparrows learn to sing by first memorizing an adult's song and then progressively matching their vocalizations to this model during plastic song. Previous studies have shown that neurons in the song-system nucleus HVC of adult sparrows respond preferentially to a bird's own song. In this study, the auditory selectivity of HVC neurons in subadult birds was examined. In young, nonsinging birds who had been song tutored, these cells responded to song stimuli, and at some recording sites had distinct preferences for one song or another. As a population, however, HVC neurons in these birds showed no preference for familiar song. They were no more likely to prefer normal tutor song to reversed tutor song or to the song of another white-crowned subspecies. By contrast, in birds producing plastic song, HVC neurons were selective for the bird's own songs, even in preference to their tutor song. Therefore, during song learning the response properties of HVC neurons appear to be dynamically modified, perhaps by auditory feedback from the bird's own vocalizations. The emergence of song selectivity during plastic song may be significant both for song learning and for song perception in adult birds. PMID- 8229197 TI - Several extracellular domains of the neural cell adhesion molecule L1 are involved in neurite outgrowth and cell body adhesion. AB - The neural cell adhesion molecule L1 is a multidomain protein that plays important roles in cell adhesion, migration, and neurite outgrowth. To analyze structure-function relationships of L1 in neurite outgrowth and cell body adhesion, we have expressed and purified a set of different fragments of the extracellular part of this glycoprotein in CHO cells and in Escherichia coli. When neurite outgrowth from small cerebellar neurons was measured on substrate coated L1 or L1 fragments, neurite outgrowth was promoted by the immunoglobulin like domains I-II, III-IV, and V-VI, and by the fibronectin type III homologous repeats 1-2, while the fibronectin type III homologous repeats 3-5 were ineffective. In contrast, cell bodies of small cerebellar neurons adhered mostly to the immunoglobulin-like domains I-II and V-VI, and to the fibronectin type III homologous repeats 3-5, but less to the immunoglobulin-like domains III-IV and fibronectin type III homologous repeats 1-2. In both assays, the neuronal cell surface receptor for all active protein fragments was identified as L1. No significant differences in functional activities were found between fragments with and without carbohydrate structures. These findings indicate that L1 uses several domains for homophilic interactions overlapping for the two functions analyzed here, but also showing some regional specialization. Furthermore, we show that a homophilic molecule uses several domains in one function, with neurite outgrowth requiring more domains than adhesion for maximal activity. PMID- 8229199 TI - The functional anatomy of limbic status epilepticus in the rat. I. Patterns of 14C-2-deoxyglucose uptake and Fos immunocytochemistry. AB - Limbic status epilepticus was induced in awake, unrestrained rats by electrically stimulating the olfactory cortex or the basal amygdaloid nucleus for about 40 min. One of four stable forms of status was induced, which were distinguished on the basis of their behavioral and EEG manifestations, and their distinct patterns of 14C-2-deoxyglucose uptake and Fos-like immunoreactivity. Type I status was characterized by sporadic EEG discharges and the activation of the amygdalohippocampal area, but had no overt behavioral manifestation. Type II status involved incessant exploratory behaviors, single EEG discharges, and the additional activation of the basal amygdaloid nucleus, some of its efferent projections, and parts of the olfactory cortex. Type III status included all of these same patterns, plus the episodic development of ictal EEG activity associated with facial and forelimb clonus, and the concurrent recruitment of the entire amygdala, ventral hippocampal formation, prefrontal, insular, and olfactory cortices, and related subcortical structures. Type IV status was characterized by generalized clonus, unremitting ictal EEG discharges, and the additional activation of most of the dorsolateral neocortex, neostriatum, and thalamus. In each case of status type I, II, or III, the same anatomical structures that displayed high levels of 14C-2-deoxyglucose uptake also contained many cells that were immunoreactive for Fos, with the exception of the parataenial and mediodorsal thalamic nuclei and the substantia nigra pars reticularis. Thus, the overall patterns of 14C-2-deoxyglucose uptake and Fos-like immunoreactivity from the same animals displayed a remarkable degree of correspondence. The major results indicate that different levels of status are related to the activation of discrete epileptogenic foci, and the capacity of such foci to interact with a distinct set of interconnected anatomical structures. It is suggested that the behavioral manifestations of limbic status epilepticus may be explained by influences of limbic structures in the ventral forebrain upon lower motor elements in the brainstem and spinal cord, without the participation of the "pyramidal" motor system. PMID- 8229198 TI - Thresholds for synaptic activation of transcription factors in hippocampus: correlation with long-term enhancement. AB - Recent studies suggest a role for rapid induction of transcription factors in stimulus-induced neuronal plasticity in the mammalian brain. Synaptic activation of transcription factors has been analyzed in the hippocampus using the long-term potentiation or enhancement (LTP/LTE) paradigm. Using this approach, several studies have identified transcription factors that are induced in hippocampal granule cells by NMDA receptor-dependent mechanisms; however, the link between long-term plasticity and activation of these genes has been called into question by reports suggesting that the thresholds for LTE and gene activation differ. To address this issue, we have used a chronic in vivo recording technique to monitor mRNA responses of several transcription factor genes to two different patterns of LTE-inducing electrical stimulation of entorhinal cortical afferents to hippocampus. One pattern consisted of 10 repetitions of a 20 or 25 msec train of pulses at 400 Hz (80 or 100 pulses total). This "10-train" pattern has been used in previous studies of LTE and produces robust synaptic enhancement lasting at least 3 d (Barnes, 1979). The other stimulation pattern consisted of 50 repetitions of a 20 msec train delivered at 400 Hz (400 pulses total), which is similar to parameters used in other studies reporting induction of c-fos in association with LTE (Dragunow et al., 1989; Jeffery et al., 1990; Abraham et al., 1992). Our results indicate that expression of zif268, monitored by in situ hybridization and immunostaining, is strongly induced by the 10-train stimulus pattern to levels similar to those induced by seizure activity. JunB mRNA levels are also modestly increased by the 10-train stimulus pattern; however, increases in JunB immunostaining were not detected. Neither c-fos nor c-jun mRNA were detectably induced by this stimulus. In contrast, the 50-train stimulus pattern resulted in a robust induction of c-fos and c-jun mRNA, in addition to zif268 and junB. Transcription factor responses to either stimulus pattern were blocked by the noncompetitive NMDA receptor antagonist MK-801. Identical transcription factor responses were observed in adult (6-12-month-old) and aged (23-26-month old) rats, suggesting that synaptic mechanisms involved in these responses are preserved in aged animals. Analysis of LTE following either the 10- or 50-train stimulus patterns revealed identical magnitudes of initial induction and decay kinetics (approximately 3 d) and indicates that the 10-train stimulus pattern is sufficient to produce maximal synaptic enhancement.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229200 TI - The functional anatomy of limbic status epilepticus in the rat. II. The effects of focal deactivation. AB - Limbic status epilepticus was induced in awake, unrestrained rats by electrically stimulating the anterior piriform cortex or the basal amygdaloid nucleus for about 40 min. As described in the preceding article (White and Price, 1993), one of four stable forms of status may be induced. Each form is characterized on the basis of its behavioral and electroencephalographic manifestations, and its distinct patterns of 14C-2-deoxyglucose uptake and Fos-like immunoreactivity. This study was directed at identifying the epileptogenic foci of the two major forms of status, types II and III, by deactivating the basal amygdaloid nucleus, ventral hippocampal formation, amygdalohippocampal area, or anterior piriform cortex during these seizure states. Infusions of the local anesthetic lidocaine, the GABA agonist muscimol, or a vehicle solution alone were made into each of these structures during ongoing type II or type III status. The major finding is that deactivation of the basal amygdaloid nucleus terminated both types of status. This indicates that the basal nucleus is primarily responsible for the generation of widespread status epilepticus activity. Deactivation of the ventral hippocampal formation did not terminate the subconvulsive levels of status, but did prevent the recurrent development of sustained seizures with facial and forelimb clonus that characterize type III status. These models of status epilepticus may be particularly important for understanding seizure mechanisms that are not dependent upon the hippocampal formation. The possible clinical relevance of these findings is discussed in relation to temporal lobe epilepsy. PMID- 8229201 TI - Two structural adaptations for regulating transmitter release at lobster neuromuscular synapses. AB - The distal accessory flexor muscle (DAFM) in the lobster (Homarus americanus) walking leg consists of 5 muscle fiber bundles. All five bundles, one proximal, one distal, and 3 medial, are innervated by one excitatory and one inhibitory motor neuron. Both neurons release more transmitter on the distal bundle than on the proximal bundle. The aim of our studies was to investigate the structural basis of this differentiation. Thin sections cut at 50 microns intervals showed a similar number of excitatory synapses on the two bundles. Freeze-fracture views of excitatory synapses showed that synapse size, active zone number per synapse, and intramembrane particle density in the postsynaptic membrane are similar proximally and distally. Active zones at synapses on the distal bundle are larger and contain about 50% more large intramembrane particles, which are thought to include the voltage-gated Ca2+ channels that couple the action potential to transmitter release, than their counterparts on the most proximal bundle. This difference in channel number appears to produce a disproportionate increase in the probability of transmitter release sufficient to account for most of the proximal-distal disparity in the amplitude of the excitatory postsynaptic potential. In contrast, staining the inhibitor for antibodies to the inhibitory neurotransmitter, GABA, showed that it forms more varicosities on the distal bundle than on the proximal bundle. Because most of the synapses are located in the varicosities, differences in synapse number likely regulate the proximal distal disparity in the amount of inhibitory transmitter released. Therefore, the regional differentiation in the amount of transmitter released in the DAFM appears to be based on two distinct mechanisms. In the inhibitor, transmitter release appears to be regulated differentially by differences in synapse number. In the excitor, transmitter release appears to be regulated differentially from a similar number of synapses by differences in active zone structure. PMID- 8229202 TI - Intracellular calcium levels and calcium fluxes in the CA1 region of the rat hippocampal slice during in vitro ischemia: relationship to electrophysiological cell damage. AB - Five minutes of oxygen and glucose deprivation (termed "in vitro ischemia") causes long-term synaptic transmission failure (LTF) in the CA1 region of the rat hippocampal slice. Dependence of LTF on cell calcium was tested by generating graded reductions in cell Ca. There was a strong correlation between the average level of exchangeable cell Ca in CA1 during ischemia, and the extent of LTF. In standard buffer, exchangeable cell Ca in CA1 increased by 35% after 3 min of ischemia and remained elevated for the entire 5 min of ischemia. Unidirectional Ca influx increased by 35% during the first 2.5 min of ischemia and remained at that level for the next 2.5 min. There were no changes in unidirectional Ca efflux during this period. Thus, the accumulation results from increased influx of Ca. Ca influx during the first 2.5 min of ischemia depended entirely on NMDA channels; it was completely blocked by the noncompetitive NMDA receptor antagonist MK-801. However MK-801 had no effect during the second 2.5 min. This inactivation of NMDA-mediated influx during ischemia appears to result from dephosphorylation. Okadaic acid increased Ca influx during the second 2.5 min of ischemia and this increase was blocked by MK-801. The ischemia-induced Ca influx during the second 2.5 min of ischemia was attenuated 25% by nifedipine (50 microM) and an additional 35% by the Na/Ca exchange inhibitor benzamil (100 microM). The AMPA/kainate antagonist DNQX had no effect on the Ca influx. Antagonists were used to relate Ca influx to LTF. Blockade of enhanced Ca entry during ischemia in standard buffer (2.4 mM Ca) had no effect on LTF, consistent with total cell Ca prior to ischemia being adequate to cause complete LTF. However, MK-801 strongly protected against LTF when the buffer contained 1.2 mM Ca, a more physiological level. MK-801 combined with DNQX prevented transmission damage in standard buffer. Thus, AMPA/kainate receptor activation contributes to ischemic damage, although not by enhancing Ca entry. PMID- 8229203 TI - Differential modulation of TTX-sensitive and TTX-resistant Na+ channels in spinal cord astrocytes following activation of protein kinase C. AB - TTX-sensitive (TTX-S) and TTX-resistant (TTX-R) Na+ currents are expressed in high densities (2-8 channels/microns2) in astrocytes cultured from neonatal rat spinal cord. The two Na+ current types differ up to 1000-fold in their TTX sensitivity and additionally have different steady-state activation (g-V) and inactivation (h infinity) curves. Expression of TTX-S and TTX-R Na+ currents is confined to morphologically distinguishable subtypes of astrocytes, allowing characterization of the two types of Na+ currents in isolation: stellate cells express TTX-S Na+ currents and flat pancake cells express TTX-R Na+ currents. Activation of protein kinase C (PKC) by phorbol 12-myristate 13-acetate (PMA) exhibited different effects on TTX-S and TTX-R Na+ currents. PMA reduced peak TTX S Na+ currents by 25-60%; in contrast, PMA potentiated peak TTX-R Na+ currents by 60-150%. These effects developed within minutes, and were typically not reversible. PMA effects were voltage dependent, and shifted steady-state Na+ current activation of TTX-R and TTX-S currents by 6 and 18 mV, respectively, but without affecting their steady-state current inactivation (h infinity). PMA treatment also changed Na+ current kinetics. TTX-R current activation (tau m) was faster and current inactivation (tau h) changed from a single- to a bi exponential after PMA exposure, suggesting that PKC phosphorylation may have activated formerly quiescent Na+ channels. In contrast, TTX-S current activation (tau m) was unchanged, and current inactivation (tau h), on average, decreased by 50% following PMA exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229204 TI - Preganglionic autonomic motor neurons display normal translocation patterns in slice cultures of embryonic rat spinal cord. AB - Phenotypic differences between somatic and autonomic motor neurons (SMNs and AMNs, respectively) may be modulated by epigenetic factors during the histogenic migrations of these cells. In order to study this problem experimentally, we have developed an in vitro, organotypic slice preparation of embryonic rat spinal cord. Our main objectives for this preparation were to determine whether in vivo patterns of motor neuronal translocations were mimicked in vitro, and, if they were, to begin to analyze such movements with experimental procedures that cannot be applied to the study of mammalian spinal cord development in vivo. Using a modification of existing organotypic slice procedures, we have shown that ChAT, an axonal surface glycoprotein and a low-molecular-weight neurofilament protein are expressed in slices cultured for up to 21 d, thus indicating that spinal neurons remained viable in vitro for relatively long periods. Most importantly, retrograde labeling and subsequent confocal microscopy have shown that the SMNs and AMNs of the slice preparations become segregated ventrodorsally into two distinct subcolumns as seen in vivo. The formation of separate AMN and SMN subcolumns appears to result from a dorsal translocation of AMNs. The fact that this cellular movement occurs in the slice preparation has allowed us to follow this phenomenon directly within the same specimen over a period of days. In addition, we have been able to observe the translocation of AMNs following the removal of their peripheral synaptic targets. The results of these experiments provide further evidence that AMNs undergo a dorsal translocation during the course of spinal cord development, and that this cellular movement may be due to an active migration. They also indicate that AMN movement is not dependent upon continual connection of these neurons with the paravertebral sympathetic ganglia. PMID- 8229205 TI - Identification and characterization of Drosophila genes for synaptic vesicle proteins. AB - Proteins associated with synaptic vesicles are likely to control the release of neurotransmitter. Because synaptic transmission is fundamentally similar between vertebrates and invertebrates, vesicle proteins from vertebrates that are important for synaptic transmission should be present in Drosophila as well. This investigation describes Drosophila homologs of vamp, synaptotagmin, and rab3 that are expressed in a pattern consistent with a function in Drosophila neurotransmission. One previously reported candidate (syb), a Drosophila homolog of the vamp or synaptobrevin proteins, has been shown to be expressed at very low levels in neurons and is most abundant in the gut. A neuronal Drosophila vamp (n syb) is described here and is localized to chromosome band 62A. Northern analysis and in situ hybridizations to mRNA indicate that the novel vamp, as well as the genes for synaptotagmin (syt) and rab3 (drab3), is expressed in the Drosophila nervous system. These genes are widely (perhaps ubiquitously) expressed in the nervous system and we have no evidence of additional neuronal isoforms of synaptotagmin, vamp, or rab3. Immunoreactivity for synaptotagmin and vamp is located in synaptic regions of the nervous system. This distribution suggests that these molecules are components of synaptic vesicles in Drosophila. The conserved structure and neuronal expression pattern of these genes indicate that they may function in processes that are required for both vertebrate and invertebrate synaptic transmission. Because of their distribution in the nervous system and because n-syb, synaptotagmin, and drab3 do not appear to be in a family of functionally redundant homologs, we predict that mutation of these genes will have a profound neurological phenotype and that they are therefore good candidates for a genetic dissection in Drosophila. PMID- 8229206 TI - Low-threshold Ca2+ current and its role in spontaneous elevations of intracellular Ca2+ in developing Xenopus neurons. AB - Amphibian spinal neurons exhibit spontaneous elevations of intracellular calcium at early stages of development. The underlying calcium influx involves high voltage-activated (HVA) currents. To begin to understand how they are triggered, we have studied the biophysical properties and developmental function of low voltage-activated (LVA) T-type calcium current of neurons cultured from the embryonic neural plate. T current was recorded from young neurons (6-9 hr in vitro) and from mature neurons (18-48 hr in vitro) using whole-cell voltage clamp. For both young and mature neurons, T current has a low threshold and is activated at membrane potentials positive to -60 mV in 2 mM extracellular calcium. The current is maximal at -35 mV with a mean peak amplitude of approximately 50 pA. Nickel blocks both LVA and HVA currents, but the former are 20-fold more sensitive. Amiloride also blocks T current selectively. T current is recorded in 87% of young neurons. This percentage drops to 67% in mature neurons after 1 d in culture and to 35% in mature neurons after 2 d in culture. There are no significant developmental changes in T current threshold, peak density, time course of activation and inactivation, and pharmacological sensitivity to blockers from 6 to 48 hr in culture. Spontaneous transient calcium elevations in young neurons assayed by fluo-3 fluorescence are blocked by nickel or amiloride at concentrations that specifically block T current. T current has the lowest threshold among other inward currents in young neurons. Moreover, mathematical simulations show that T current lowers the threshold of the action potential by 15 mV. We conclude that T current can depolarize cells and trigger action potentials, and constitutes part of the cascade of events leading to spontaneous elevations of intracellular calcium in cultured neurons at early stages of differentiation. PMID- 8229207 TI - Fine-structural alterations and clustering of developing synapses after chronic treatments with low levels of NMDA. AB - In the visual pathway of frogs it is possible to apply low levels of NMDA chronically to the optic tectum and study the mechanisms underlying the stabilization of synapses developing within the CNS. Earlier studies (Cline and Constantine-Paton, 1990) found that chronic NMDA treatment of tecta innervated by two retinas results in a reduction of branching within the terminal arbors of retinal ganglion cells (RGCs). We now report that this same chronic NMDA treatment produces fine-structural changes in synaptic morphology as well as local synaptic rearrangements within the retinotectal neuropil. Chronic NMDA treatment of doubly innervated tecta was associated with a thickening or darkening of both pre- and postsynaptic densities. These changes in synapse morphology were restricted to the superficial neuropil of tecta in regions where reductions in branches of RGC axonal arbors were observed at the light microscopic level. The fine-structural effects were absent from similarly treated tecta innervated by only one eye, where RGC axonal arbor pruning was not observed. Stereological analyses indicated that the incidence of two or more presynaptic profiles converging on the same postsynaptic process was significantly increased in the NMDA-treated, doubly innervated tecta. This observed increase in synaptic clustering was not associated with a larger synaptic active zone, or with an increase in the number of synapses per unit volume. These data are discussed in the context of the hypothesis that chronic NMDA treatment raises the threshold for synapse stabilization in tectal neurons, causing the selective loss of poorly correlated synapses of both retinal and non retinal origin from tectal neuropil that is innervated by two retinas: increased pre- and postsynaptic thickening could reflect greater efficiency in the remaining synaptic contacts and their closer spatial proximity on the same postsynaptic process is consistent with greater cooperativity and less competition. PMID- 8229208 TI - Neurotrophin-4/5 is a mammalian-specific survival factor for distinct populations of sensory neurons. AB - We have studied the effect of human recombinant neurotrophin-4/5 (NT-4/5) on the survival of developing PNS neurons from embryonic mice and chickens. NT-4/5 transiently supported mouse NGF-dependent trigeminal and jugular neurons at early stages of target field innervation and mouse brain-derived neurotrophic factor (BDNF)-dependent no-dose neurons during the phase of naturally occurring cell death. NT-4/5 was as potent as BDNF in supporting the survival of these neuronal populations. Surprisingly, NT-4/5 was 3 orders of magnitude less potent than BDNF as a survival factor for early chick dorsomedial trigeminal sensory neurons and did not support the survival of chick BDNF-dependent trigeminal mesencephalic or ventrolateral trigeminal sensory neurons at any of the developmental stages tested. Thus, NT-4/5 is a survival factor for certain embryonic mouse cranial sensory neurons. It is the first species-specific neurotrophin to be identified and it can discriminate at high concentrations between different BDNF-responsive chick neurons. PMID- 8229209 TI - Protein tyrosine phosphatases expressed in the developing rat brain. AB - Previous studies of the developing nervous system have shown that cell-cell and cell-matrix interactions are involved in a variety of processes such as the proliferation, migration, and differentiation of neurons. While many cell-surface molecules have been identified, the signal transduction mechanisms through which they modify cellular responses are poorly understood. Recent studies have described a new and large family of enzymes, protein tyrosine phosphatases (PTPases), that may play a key role in transduction of cell surface events. Opposing the actions of protein tyrosine kinases (PTKs), PTPases can determine the state of tyrosine phosphorylation of a protein and regulate its function. Within the family of PTPases, two subgroups have been characterized: low molecular-weight cytoplasmic (nonreceptor) PTPases and high-molecular-weight transmembrane (receptor) PTPases. Many receptor PTPases have fibronectin type III and/or Ig-like domains in their extracellular domains, suggesting that they have dual functions: cell adhesion and signal transduction. Such molecules may play a role in cellular recognition events that mediate the accurate assembly of the nervous system. Using polymerase chain reaction with degenerate primers and a neonatal rat cortex cDNA library, we have identified a number of putative PTPase domains expressed in brain. Three are characterized here. These three sequences are most abundantly expressed in the developing cortex and so are named cortex enriched protein tyrosine phosphatases (CPTPs) 1, 2, and 3. CPTP1 and CPTP3 show sequence homology to receptor PTPases and detect multiple high-molecular-weight mRNAs that are expressed preferentially in the developing CNS. Analysis of a longer cDNA indicates that CPTP1 and CPTP3 are the first and second phosphatase domains of a single receptor PTPase. CPTP2 identifies a single, smaller mRNA species with sequence homology to nonreceptor PTPases. Within the CNS, mRNAs detected by all three CPTPs are expressed at highest levels during prenatal and early postnatal days and are downregulated in the adult. In situ hybridization demonstrates that the CPTPs are expressed by progenitor cells and developing neurons. The spatial and temporal regulation of CPTPs suggests that they may play a role in neuronal development. PMID- 8229210 TI - Primary sensory neurons express a Shaker-like potassium channel gene. AB - Developmentally regulated action potentials are a hallmark of Rohon-Beard cells, a class of sensory neurons. In these neurons as well as other primary spinal neurons of Xenopus laevis, the functional differentiation of delayed-rectifier potassium current regulates the waveform of the action potential during the initial day of its appearance. Later, the acquisition of another voltage dependent potassium current--the A current--plays a major role in regulating excitability. In order to understand the molecular basis of this functional differentiation, genes encoding voltage-dependent potassium currents expressed in the embryonic amphibian nervous system are being cloned. Here, we report the functional properties and developmental localization of a second Xenopus Shaker like gene (Xenopus Kv 1.1; XSha1; GenBank accession number M94258) encoding a potassium current. Homology screening with the mouse gene MBK1 led to its isolation. Functional expression in oocytes identifies it as a delayed-rectifier current when assembled as a homooligomeric structure. Specific transcripts corresponding to XSha1 and to the previously cloned gene XSha2 are both detectable by RNase protection in RNA isolated from the embryonic nervous system. However, whole-mount in situ hybridization reveals the temporal pattern and cellular localization of XSha1 but not XSha2 mRNA, suggesting that the concentration of XSha2 transcripts in individual cells is lower than the threshold for detection by this method. Of particular interest, Rohon-Beard cells express XSha1 mRNA. In addition, XSha1 mRNA is detected in several structures containing neural crest derivatives including spinal ganglia, the trigeminal ganglion, and branchial arches; its presence in motor nerves and lateral spinal tracts suggests that both CNS and PNS glia express the mRNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229211 TI - Relative properties and localizations of synaptic vesicle protein isoforms: the case of the synaptophysins. AB - Synaptophysins are abundant synaptic vesicle proteins present in two forms: synaptophysin, also referred to as synaptophysin I (abbreviated syp I), and synaptoporin, also referred to as synaptophysin II (abbreviated syp II). In the present study, the properties and localizations of syp I and syp II were investigated to shed light on their relative functions. Our results reveal that syp II, similar to syp I, is an abundant, N-glycosylated membrane protein that is part of a heteromultimeric complex in synaptic vesicle membranes. Cross-linking studies indicate that syp II is linked to a low-molecular-weight protein in this complex as has been observed before for syp I. Furthermore, after transfection into CHO cells, syp II, similar to syp I, is targeted to the receptor-mediated endocytosis pathway. Immunocytochemistry of rat brain sections reveals that syp II expression is highly heterogeneous, with high concentrations of syp II only in selected neuronal populations, whereas syp I is more homogeneously expressed in most nerve terminals. In general, nerve terminals expressing syp II also express syp I. In addition to high levels of syp II observed in selected neurons, a rostrocaudal gradient of syp II expression was observed in the cerebellar cortex. Immunoelectron microscopy confirmed that syp II is localized to synaptic vesicles. Immunoprecipitations of synaptic vesicles from rat brain with antibodies to syp I demonstrated that syp II is colocalized with syp I on the same vesicles. However, after detergent solubilization, no coimmunoprecipitations of the two proteins were observed, suggesting that they are not complexed with each other although they are on the same vesicles. Together our results demonstrate that syp I and syp II have similar properties and are present on the same synaptic vesicles but do not coassemble. The presence of the two proteins in the same nerve terminal suggests that they have similar but nonidentical functions and that the relative abundance of the two proteins may contribute to the functional heterogeneity of nerve terminals. PMID- 8229212 TI - Carpal tunnel syndrome: treatment and rehabilitation therapy for the dental hygienist. PMID- 8229213 TI - The important role of dental hygienists in the identification of child maltreatment. PMID- 8229214 TI - Children and bereavement: the dental hygienist's role. PMID- 8229215 TI - Oral healthcare and cultural barriers. PMID- 8229216 TI - The ADHA framework for theory development: a white paper requested by the 1992 house of delegates. PMID- 8229217 TI - The multicultural knowledge of registered dental hygienists: a pilot study. AB - PURPOSE: In a 1990 report to the nation (Healthy People 2000), the United States Public Health Service identified members of certain racial and ethnic minority groups as experiencing above average incidences of caries, periodontal disease, and oral cancer. To help improve healthcare offered these populations, it is now recommended that it be planned and presented in a manner that is compatible with their existing cultural values and beliefs. The purpose of this study was to examine (1) dental hygienists' knowledge of the values, beliefs, lifestyles, and health practices of four ethnic minority groups--African Americans, American Indians, Asian Americans, and Hispanic Americans; and (2) the effects of dental hygienists' age, education, and amount of professional experience level on such knowledge. METHODS: A total of 30 registered dental hygienists completed a 20 item Knowledge of Cultures Test and a demographic questionnaire during the summer of 1991. The data were analyzed by means of the independent t-test. RESULTS: The findings indicate that the respondents possess a low level of multicultural knowledge of the values, beliefs, lifestyles, and health practices of ethnic minority groups. The dental hygienists' age, education, and amount of professional experience had no significant effects on the level of multicultural knowledge. CONCLUSIONS: The findings suggest that curricula within dental hygiene programs need to be planned and taught from a transcultural perspective to help students develop the knowledge necessary to plan beneficial care for clients from ethnic minority groups. PMID- 8229218 TI - Pennsylvania dental hygienists' knowledge, attitudes, and infection control practices in relation to AIDS and AIDS patients. AB - PURPOSE: Limited documentation is found on dental hygienists' attitudes toward acquired immunodeficiency syndrome (AIDS) patients and their knowledge and practice of clinical infection control to prevent disease transmission to themselves and their patients. The purpose of this paper was to survey practicing Pennsylvania dental hygienists to document 1) their infection control practices; 2) their attitudes towards AIDS patients; and 3) their knowledge of clinical infection control practices. METHODS: A self-administered survey questionnaire, with fixed-alternative responses, was mailed in January 1991 to a random sample (N = 300) of licensed, practicing Pennsylvania dental hygienists. The questionnaire comprised eight multiple-choice questions for demographic purposes and 89 Likert-type questions eliciting information in five areas: AIDS-related knowledge, attitudes toward AIDS patients, knowledge of recommended Centers for Disease Control and Prevention (CDC) infection control measures, individual infection control measures, and individual laboratory infection control measures. Means and modes for individual questions and for specific categories were determined and analyzed utilizing Spearman rho correlation coefficients (p < .05). Mean scores were also tabulated for actual operator/laboratory infection control practices for both a routine patient and an AIDS patient. Those scores were analyzed utilizing the Wilcoxen signed-ranks test. RESULTS: Two hundred twenty questionnaires were returned for a 73.3% initial response rate. One hundred fifty-four of those returned were usable, for a 64% response rate. Results indicated that 94.2% of surveyed dental hygienists had comprehensive knowledge about AIDS and 92% had comprehensive knowledge of CDC-recommended infection control procedures. Eighty-five percent of respondents possessed a moderate or high feeling of worry concerning treatment of AIDS patients. The majority of surveyed dental hygienists routinely practiced the use of glasses, masks, and gloves; the use of disposable items; and surface disinfection of light handles, instrument bracket trays, and patient chair switches. Knowledge of recommended infection control procedures for dentistry was found to be associated (r = .22) with adherence to recommended infection control practices. Accurate knowledge about AIDS showed a weak (r = -.088) and nonsignificant relationship with dental hygienists' attitudes toward AIDS patients. Use of recommended infection control practices was found to be associated (r = -.20) with less fear concerning the treatment of AIDS patients. In addition, dental hygienists' infection control practices varied according to their perception of patient HIV status. CONCLUSIONS: Since Pennsylvania dental hygienists, within the limitations of this study, appear not to follow CDC guidelines on proper and responsible operatory/laboratory aseptic techniques stringently, and to differentiate infection control procedures based on perceived patient HIV status, recommendations are that 1) the Commonwealth of Pennsylvania should mandate that all Pennsylvania-licensed dental hygienists take at least one state-approved course on operatory/laboratory infection control every two years to qualify for relicensure; 2) all dental and dental hygiene education institutions and professional organizations should place more emphasis on strict adherence to the various agency recommended clinical guidelines for infection control; and 3) all dental hygienists should continually strive to update their own knowledge of current infection control practices. PMID- 8229219 TI - Human immunodeficiency virus-associated periodontal diseases: a review. PMID- 8229220 TI - Tuberculosis: a reemerging hazard for oral healthcare workers. PMID- 8229221 TI - Samarium-153-EDTMP in bone metastases of hormone refractory prostate carcinoma: a phase I/II trial. AB - Samarium-153-ethylenediaminetetramethylene phosphoric acid (EDTMP), a bone seeking radiopharmaceutical, was given to prostate cancer patients in a dose escalation protocol for pain palliation to determine the maximally tolerated dose. Fifty-two patients with hormone refractory prostate cancer with bony metastases were treated with doses beginning at 0.5 mCi/kg (18.5 MBq/kg), escalating in 0.5-mCi (18.5 MBq) increments to 3.0 mCi/kg (111 MBq/kg). Pain response after treatment was assessed as well as hematologic and serum chemistry parameters. Pain palliation with a mean duration of 2.6 mo was present in 74% of the patients. Toxicity was exclusively hematologic at the highest dose levels. No infectious or bleeding complications occurred, with 45 of the 52 (86%) patients demonstrating complete hematologic recovery. Patients receiving higher doses had significantly greater reductions in serum prostate specific antigen and serum prostatic acid phosphatase levels. The patients receiving greater doses also showed a trend toward improved survival. PMID- 8229222 TI - Effect of motion on thallium-201 SPECT studies: a simulation and clinical study. AB - Although patient motion on 201Tl SPECT studies has been reported as a source of artifacts, systematic studies on motion patterns and resultant artifacts are lacking. Accordingly, we simulated 74 motion patterns upon a normal study. The tomograms were assessed for presence of defects: The "motion pixel area index" ranged from 1 to 83; 26 of 30 (87%) simulations with an index > or = 21 had defects, whereas 38 of 44 (86%) simulations with an index < 21 were normal. Defect location was dependent on motion direction; defect intensity was dependent on its magnitude and timing. Review of data acquisition in 164 recent normal patient studies revealed motion in 42 (26%). Motion was generally minimal and caused defects in only seven (4%). Thus, mild motion is unlikely to produce defects. In our laboratory, motion is now an infrequent source of artifacts; severe motion produces recognizable patterns that depend on its direction, magnitude and timing. PMID- 8229223 TI - Detailed analysis of patients with matched ventilation-perfusion defects and chest radiographic opacities. AB - We performed a retrospective evaluation of the data obtained from the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) study to determine the prevalence and distribution of pulmonary embolism (PE) in patients with matching ventilation-perfusion (V/Q) defects and chest radiographic opacities (triple matches). From the 1,487 patients in whom diagnostic V/Q scans were obtained, 247 (17%) patients were recorded as having matching V/Q defects and chest radiograph opacities in at least one lung zone. Diagnostic angiograms were available for 275 lung zones (233 patients) which demonstrated matching V/Q defects and chest radiograph opacities. The overall prevalence of PE in all lung zones with triple matches was 26%. The prevalences of PE in the upper, middle and lower lung zones were 11%, 12% and 33%, respectively. Pulmonary embolism was significantly more common in lower lung zone triple matches compared with the upper and middle lung zone triple matches (p < 0.005). There was no correlation between the size of the V/Q and chest radiographic abnormalities and the prevalence of PE. We conclude that in patients with matching V/Q defects and chest radiographic opacities isolated to the upper and middle zones the V/Q scan can be interpreted as representing a low probability of PE. Similar findings in the lower lung zones represent an intermediate probability of PE. The application of this modification to V/Q scan interpretation criteria will aid in reducing the number of intermediate V/Q lung scan interpretations. PMID- 8229224 TI - Abnormalities on ventilation/perfusion lung scans induced by bronchoalveolar lavage. AB - Studies were performed before and at varying times after lavage in 10 normal volunteers to assess whether bronchoalveolar lavage results in significant abnormalities on ventilation/perfusion lung scans and chest x-rays. Abnormal lung scans were obtained in six subjects, interpretable as intermediate (three scans), low (one scan) and very low (two scans) probability for pulmonary emboli. Defects varied from multisegmental to subsegmental in size, while chest x-rays were normal in all but one. Both the extent and frequency of defects tended to decrease with time; 24 hr after bronchoalveolar lavage only one of four subjects had a minimally abnormal scan. It is recommended that ventilation/perfusion lung scanning be delayed at least 24 hr following bronchoalveolar lavage to avoid problems in interpretation of defects which may merely be the result of the lavage. PMID- 8229225 TI - Controlled delivery of krypton-81m boli in normal subjects: results and implications. AB - Interregional sequential filling potentially affects lung ventilation imaging, depending on the distribution of the tracer within the inspired volume. We investigated its importance quantitatively under near tidal breathing conditions in the upright lung using a short-lived radioactive tracer. Ten normal volunteers performed two runs of 900-ml breaths (from functional residual capacity) in which 100 ml of 81mKr boli were delivered "early" or "late" in inspiration, i.e., 50 ml or 450 ml volumetric depth. Apex-to-base gradients in the vertical profile were 106 +/- 22 (s.e.) counts/cm (early) and -187 +/- 24 (s.e.) counts/cm (late). Ratios of upper-to-lower regional ventilation (U/L) were 0.88 +/- 0.01 (s.e.) (early) and 0.81 +/- 0.01 (s.e.) (late). Simulations with a compartment model show that a simple pattern of sequential filling can by itself account for the experimental results observed. Control over 81mKr delivery can be important to physiologically accurate assessment of ventilation-perfusion matching. Controlled delivery techniques could also modify effectiveness and targeting of other inhaled agents including therapeutic aerosols. PMID- 8229226 TI - Cholescintigraphy in the diagnosis of acute cholecystitis: morphine augmentation is superior to delayed imaging. AB - Morphine-augmented radionuclide hepatobiliary imaging has been used as an alternative to delayed imaging for the diagnosis of acute cholecystitis. Previous studies have indicated that the morphine-augmentation is as useful as, or more useful than, delayed imaging. A careful comparison of the efficacy of the two techniques appears warranted because: (1) most early studies did not compare the efficacy of the two techniques in a single report using comparable patient populations; (2) the reported efficacy of morphine-augmentation is based primarily on study designs which excluded cases of early gallbladder visualization without morphine, while most delayed imaging protocols included these cases; and (3) there were concerns about the potential consequences of a false-negative morphine examination. This study compared the efficacy of morphine augmentation with delayed imaging in those cases in which the gallbladder was not visualized during the first hour of study. Of 306 consecutive patients who were scanned to rule out acute cholecystitis, the gallbladder was visualized within 1 hr in 215 cases. In the remaining 91 cases, 46 patients had delayed imaging (17 true-positive, 10 true-negative, 19 false-positive and 0 false-negative), and 45 had morphine-augmentation (24 true-positive, 15 true-negative, 4 false-positive and 2 false-negative). The data indicate that delayed imaging has a significantly lower specificity and positive-predictive value for acute cholecystitis than morphine-augmentation and a slightly higher (statistically insignificant) sensitivity and negative-predictive value. These results appear to be supported by a reanalysis of the data that has already been reported in the literature. PMID- 8229227 TI - Left ventricular ejection fraction assessed from gated technetium-99m-sestamibi SPECT. AB - By means of ECG gating of tomographic (SPECT) 99mTc-sestamibi (MIBI) images, myocardial perfusion and wall thickening have been evaluated after a single tracer injection. To determine if left ventricular ejection fraction (LVEF) can also be measured from gated MIBI SPECT, 30 patients 1 wk to 6 mo after myocardial infarction (MI) received 22-30 mCi 99mTc-MIBI during treadmill exercise. Eight frame per cardiac cycle gated MIBI 180 degrees SPECT was performed 60 min thereafter. Using 6.4-mm thick mid-ventricular vertical and horizontal long-axis slices from R-wave triggered end-diastolic and end-systolic frames, two independent observers manually drew endocardial borders at a count level of 34% of the maximum. LVEF was computed by the Simpson's rule method, corrected for the average point spread function of the SPECT camera. Results were correlated with LVEF determined from planar gated 99mTc-blood-pool studies performed within 4 days. LVEFs calculated from gated MIBI SPECT ranged from 0.21 to 0.73 and correlated linearly with gated blood-pool values (correlation coefficients ranged from 0.79 to 0.88; interobserver variability r = 0.75; intraobserver reproducibility r = 0.75). We conclude that in patients with MI resting LVEF can be determined from gated MIBI SPECT, thereby considerably augmenting the technique's diagnostic and prognostic value. PMID- 8229228 TI - Left ventricular peak power during exercise: a noninvasive approach for assessment of contractile reserve. AB - Cardiac peak power, a contractility index based upon instantaneous changes in intracavitary pressure and systolic peak flow, was measured at rest and during supine exercise in 26 patients with coronary artery disease and 8 healthy subjects. The pathophysiological significance of this index was compared with left ventricular ejection fraction (LVEF) during exercise. Cardiac peak power, ejection fraction, end-diastolic volume, stroke volume, cardiac output and systemic vascular resistance were measured at rest, during three stages of supine bicycle ergometry and two stages of recovery. Cardiac peak power increased continuously in healthy subjects, from 5.4 +/- 0.8 W/ml at rest to 11.4 +/- 3.1 W/ml at peak exercise, p < 0.001. In patients, peak power increased initially, reached a plateau in stage 2, and subsequently remained unchanged in stage 3 (5.6 +/- 2 versus 5.6 +/- 1.6 W/ml, p = ns). Ejection fraction demonstrated a flat response during exercise in patients, contrasting with a 42% increase in cardiac peak power. The lack of increase in ejection fraction was attributed to its dependence on afterload. Peak power showed no correlation with systemic vascular resistance (r = 0.01, p = ns). In a subgroup of patients with low resting LVEF (LVEF = 26% +/- 7%), peak power increased 70% during exercise, from 2.0 +/- 0.7 to 3.5 +/- 1.7 W/ml, p < 0.05, in contrast to a flat ejection fraction response. Thus, cardiac peak power, a relatively afterload-independent index of left ventricular performance and contractility can be obtained noninvasively during exercise. PMID- 8229229 TI - PET imaging of non-small-cell lung carcinoma with carbon-11-methionine: relationship between radioactivity uptake and flow-cytometric parameters. AB - Carbon-11-methionine PET scans were obtained from 24 patients with non-small-cell lung carcinoma for whom surgical treatment was considered. The tumor mass was visualized with clear delineation. After PET scanning, the tumor was removed by lobectomy or pulmonectomy. The tumor tissue was first processed to yield tumor cell suspensions and then subjected to DNA flow cytometry. Comparison between 11C uptake rate and flow-cytometric data gave the following results: 11C uptake rate in the tumor correlated well with the cellular DNA content (DNA index) of tumor cells at the resting state of cell division (G0 + G1-phase) (r = 0.67). The correlation between 11C uptake rate and S-phase cell percentage was markedly high (r = 0.76), and the correlation between 11C uptake rate and S+G2/M-phase cell percentage was extremely high (r = 0.86). It was concluded that the tumor uptake rate of 11C-methionine was representative of tumor growth rate in this tumor type. PMID- 8229230 TI - Clinical evaluation of interictal fluorine-18-fluorodeoxyglucose PET in partial epilepsy. AB - Interictal [18F]fluorodeoxyglucose (FDG) positron emission tomography (PET) is useful in presurgical evaluation of medically refractory partial epilepsies. Limited replicability of image interpretation may restrict this application. We investigated interpretation replicability in 241 18F-FDG studies performed with three different tomographs in partial epilepsy patients. Two investigators independently interpreted the studies with a standardized evaluation protocol and without knowledge of the subjects. Replicability of these unbiased interpretations in detection of regional hypometabolism was best for studies performed with the highest performance tomograph. Interictal 18F-FDG studies performed with this tomograph revealed regional hypometabolism in 62% of patients who had normal cerebral magnetic resonance imaging (MRI). Replicability of interpretations in detecting regional hypometabolism was adequate for clinical application of interictal 18F-FDG studies performed with any of the tomographs. PMID- 8229231 TI - Assessment of coronary reserve in man: comparison between positron emission tomography with oxygen-15-labeled water and intracoronary Doppler technique. AB - This study compared positron emission tomography (PET) using oxygen-15-labeled water for measurement of coronary reserve with intracoronary Doppler in patients with left anterior descending artery stenosis and patients with no coronary lesion and a coronary reserve 3 as assessed by the invasive technique. To determine whether PET measurement of coronary reserve is altered by partial volume effect, patients with left ventricular dysfunction due to idiopathic cardiomyopathy were studied with both techniques. Direct ultrasonic measurement of coronary reserve was performed the day prior to the PET study: a Doppler catheter was placed in the proximal left anterior descending artery; mean velocity was recorded at baseline and after dipyridamole administration. Using a time-of-flight PET system, patients underwent: (1) an intravenous bolus of oxygen 15-labeled water at baseline and 4 to 6 min after intravenous infusion of dipyridamole using the same protocol as for Doppler study and (2) a 18F fluorodeoxyglucose (FDG) myocardial imaging. Oxygen-15 time-activity curves were recorded in myocardial regions of interest (ROIs) drawn on a static FDG image. Using the left ventricular time-activity curve as an input function, a standard model with a single-tissue compartment was fitted to the PET data; myocardial blood flow was estimated as the blood-to-tissue transfer rate constant. Coronary reserve measured by PET was well correlated with the measured by intracoronary Doppler (r = 0.98, p < 0.001 for global population). This PET method is an accurate and reliable tool to noninvasively measure coronary reserve in patients, even in those with left ventricular dysfunction. PMID- 8229232 TI - Comparison of methods for identifying early methotrexate-induced hepatotoxicity in patients with rheumatoid arthritis. AB - Hepatotoxicity may complicate therapy with methotrexate in patients with rheumatoid arthritis. Prevention of cirrhosis may depend upon early identification of liver damage, usually accomplished by serial biopsy. To determine the adequacy of noninvasive methods for identifying hepatotoxicity, 22 sets of data were obtained in patients undergoing therapy with methotrexate for rheumatoid arthritis. Comparisons were made between liver biopsy, hepatocellular enzymes and two noninvasive radioisotopic methods that have been shown to be abnormal in hepatocellular disease: the rate constant of excretion of the 14C aminopyrine and the time from injection to peak hepatic activity of 99mTc diisopropylimidodiacetic acid. The hepatocellular enzymes and the time-to-peak activity of diisopropylimidodiacetic acid were not useful predictors of methotrexate-induced hepatotoxicity. The aminopyrine breath test was abnormal in approximately half the patients with hepatotoxicity but showed poor specificity. Noninvasive methods remain inferior to biopsy for the detection of mild to moderate methotrexate-induced hepatotoxicity in patients with rheumatoid arthritis. PMID- 8229233 TI - Biodistribution and dosimetry of iodine-123-IBF: a potent radioligand for imaging the D2 dopamine receptor. AB - Iodine-123-labeled iodo-benzofuran (IBF) is a potent D2 dopamine receptor antagonist that has been developed as a potential SPECT imaging agent. This report documents its biodistribution and radiation dosimetry in seven healthy humans. Approximately 100 MBq of IBF were administered to each volunteer. Urine was collected to measure the fraction of the activity that was voided by the renal system. Conjugate images were serially acquired over 24 hr to determine the fraction of activity in the other organs. Standard image analysis techniques were used to measure the geometric mean count rates in the brain, GI tract, heart, liver, lungs and thyroid at each time point. Corrections for attenuation were made with 123I transmission scans. Multicompartmental modeling was used to stimulate and predict the biokinetic behavior of 123I-IBF in the rest of the body. The absorbed doses for 24 organs were then estimated with the MIRD formalism. Rapid biological washout minimized the absorbed dose to most tissues. The excretory organs were exposed to the most radiation. The lower large intestine received about 0.13 mGy/MBq (0.48 rad/mCi), and the urinary bladder received 0.11 mGy/MBq. This low radiation burden will allow more than 370 MBq (10 mCi) to be administered to healthy research subjects during each study of the D2 receptor. Since high quality images of the brain can be obtained with half this amount, the findings suggest that 123I-labeled IBF has a large margin of radiation safety in humans. Its stability in vivo and its high target-to background contrast ratio in the human brain may make it a useful SPECT imaging agent. PMID- 8229234 TI - Usefulness of gallium-67-citrate scans in patients with acute disseminated tuberculosis and comparison with chest x-rays. AB - Sixteen patients (4 females, 12 males; ages: 15-80 yr) were admitted to our hospital and diagnosed as having acute disseminated tuberculosis (TB). Gallium-67 citrate scans and chest x-rays were used to localize the TB. The results revealed that: (1) most patients had an underlying disease (malignancy or immunocompromise); (2) gallium lung scans demonstrated multiple patterns from a negative picture to greater than liver uptake (localized or diffuse); (3) the extrapulmonary TB focus could be clearly visualized by whole-body gallium scans; (4) most chest x-rays had a miliary pattern and few cases were negative or localized. A combination examination, including gallium scans and chest x-rays, is needed to avoid missing early diagnosis of acute disseminated tuberculosis, especially in high risk group patients. PMID- 8229235 TI - Optimized diagnostic strategy for neuroblastoma in opsoclonus-myoclonus. AB - Infantile myoclonic encephalopathy (opsoclonus-myoclonus or IME) is a rare clinical syndrome associated with occult neuroblastoma in 20%-50% of all cases. IME is the initial presentation of neuroblastoma in 1%-3% of children. Imaging approaches including chest radiography and abdominal computed tomography (CT) have been proposed to detect neuroblastoma in IME. Metaiodobenzylguanidine (MIBG) is highly effective in the detection of neuroblastoma. These scans can identify both soft-tissue and skeletal lesions anywhere in the body. Our purpose was to attempt to determine the best screening method for detection of occult neuroblastoma in patients with IME. Records of all neuroblastoma patients from 1983 to May 1991 were reviewed. Four cases of IME with neuroblastoma were identified in which imaging studies included an MIBG scan. All four patients had positive MIBG scans (100%) while only two had masses on initial CT (50%). In the three patients initially evaluated by traditional methods, the mean time to diagnosis and the mean number of advanced radiologic studies were 7.5 mo and 7.3 studies respectively. The patient screened with MIBG had only cranial and abdominal CT prior to surgery. Although based on a limited number of patients, results suggest that MIBG may prove to be a useful screening procedure in patients with IME. Traditional imaging modalities can then be directed to evaluate sites of disease identified by MIBG scans. PMID- 8229236 TI - Assessment of the efficacy of iodine-131 for thyroid ablation. AB - It is customary to ablate residual tissue after near-total thyroidectomy for thyroid carcinoma by administering 131I. A recent trend has been to use lower 131I doses. This study was designed to assess the efficacy of thyroid ablation by 1110 MBq of 131I (30 mCi) in patients who had near-total thyroidectomy for papillary, mixed or follicular thyroid carcinoma. Four months after surgery, a whole-body scan was done using 185 MBq (5 mCi) of 131I after withdrawal of L thyroxine for 5-6 wk. Residual thyroid area was then measured by planimetry of the thyroid scan. Patients received ablation therapy within 5 days after scanning and one or more subsequent scans were performed 6 mo later. Forty-four patients were treated to ablate residual functional thyroid tissue. Of these, 12 (27%) had successful ablation. Total body areas (1.63 +/- 0.16 versus 1.83 +/- 0.30, p < 0.03) and residual thyroid tissue (1.4 +/- 1.4 versus 2.0 +/- 1.2 cm2, p < 0.05) were less in patients with total thyroid ablation while there was a trend for a smaller incidence of associated goiter in those patients (1/12 versus 13/32, p < 0.07). Nine of the 17 (53%) patients with a total body area less than 1.9 m2 and/or with a residual thyroid tissue less than 2.1 cm2 and/or without associated previous associated diffuse or multinodular goiter had a total thyroid ablation, while 3 of the 27 (11%) patients who did not have these characteristics had a successful therapy (p < 0.005). Our data suggest that 1110 MBq (30 mCi) of 131I can achieve total ablation of residual thyroid tissue after near-total thyroidectomy particularly in patients with lower total body area and smaller residual thyroid tissue without associated previous diffuse or multinodular goiter. PMID- 8229237 TI - Detection of postural cerebral hypoperfusion with technetium-99m-HMPAO brain SPECT in patients with cerebrovascular disease. AB - Seventeen of 19 patients (67 +/- 8 yr, 17 males and 2 females) had more than 75% unilateral stenosis or occlusion of the internal carotid or middle cerebral artery and two patients had carotid endarterectomy that previously had 90% stenosis of the internal carotid artery. They were studied during upright 99mTc HMPAO brain SPECT. HMPA was injected immediately after arising from a supine position. Patients were classified into Group A (n = 10) with occlusion of the internal carotid or the middle cerebral artery or Group B (n = 9) with more than 75% unilateral stenosis of the internal carotid or the middle cerebral artery and with carotid endarterectomy. Additional cerebral blood flow perfusion abnormalities between upright and supine 99mTc-HMPAO brain SPECT were detected in seven patients in Group A and in only one patient in Group B. Semiquantitative analysis showed that the asymmetric ratios between upright and supine positions changed significantly in Group A from 0.82 +/- 0.15 to 0.89 +/- 0.10 (p < 0.01), but not in Group B, from 0.89 +/- 0.11 to 0.92 +/- 0.12 (ns). Additional perfusion abnormalities were relevant to occlusion of the internal carotid or middle cerebral artery indicated postural cerebral hypoperfusion. We conclude that upright 99mTc-HMPAO brain SPECT visualizes postural cerebral hypoperfusion possibly related to silent cerebral ischemia. PMID- 8229238 TI - Re-evaluation of amino acid PET studies: can the protein synthesis rates in brain and tumor tissues be measured in vivo? AB - The significance of L-[methyl-11C]methionine (11C-Met), L-[1-11C]leucine (11C Leu) and L-2-[18F]fluorotyrosine (18F-Tyr) for measuring protein synthesis rates in the brain and in tumors by PET was re-evaluated. Tissue uptake and protein incorporation of 3H-Met, 14C-Leu and 18F-Tyr were investigated in mice bearing the FM3A mammary carcinoma. In the control group, the uptake of all three tracers in the brain and FM3A and their incorporation into the acid-precipitable fraction (APF) increased over 60 min. When the protein synthesis in vivo was inhibited by cycloheximide, the incorporation of all three tracers into the APF was significantly reduced to 6%-32% and 3%-11% of the control in the brain and FM3A, respectively. Under these conditions, total uptake of 14C-Leu in the brain and FM3A decreased rapidly, and most of the 14C in the APF was detected as proteins. On the other hand, 3H-Met and 18F-Tyr uptake continued to increase, and significant amounts of radioactivity were incorporated into nonprotein materials. In mice given ouabain to inhibit amino acid transport, total uptake of all three amino acids by FM3A was reduced to 67%-74% of the control 5 min postinjection. These results demonstrate that uptake of the three amino acids is affected by alterations in the amino acid transport system in the brain and tumor tissues, but that only 14C-Leu uptake reflects protein synthesis rates. PMID- 8229239 TI - Carbon-11-forskolin: a ligand for visualization of the adenylate cyclase-related second messenger system. AB - To visualize the adenylate cyclase (AC)-related second messenger system, [11C]forskolin, [11C]1-acetyl-7-deacetylforskolin, [11C]1,9-dideoxyforskolin and [11C]1-deoxyforskolin were synthesized by acetylation of the respective deacetyl precursors using [11C]acetylchloride and dimethylaminopyridine. The radiochemical yield of [11C]forskolin, [11C]1-acetyl-7-deacetylforskolin, [11C]1,9 dideoxyforskolin and [11C]1-deoxyforskolin calculated from trapped [11C]CO2 were 5%, 10%, 15% and 18%, respectively. Since the 1- and 9-OH groups on the forskolin structure are critical for specific binding to AC (active type), we considered [11C]1-acetyl-7-deacetylforskolin, [11C]1,9-dideoxyforskolin and [11C]1 deoxyforskolin to be nonspecific forskolin analogs. A comparative study of [11C]forskolin and its analogs on the n-octanol/phosphate buffer (pH 7.4) partition ratio showed that [11C]1-acetyl-7-deacetylforskolin has similar physical properties to [11C]forskolin. In the mouse heart, kidneys, liver and lungs, more [11C]forskolin accumulated than [11C]1-acetyl-7-deacetylforskolin. Moreover, simultaneous [11C]forskolin with forskolin (10 micrograms) administration reduced the accumulation of [11C]forskolin particularly in the heart to the level of [11C]1-acetyl-7-deacetylforskolin. These results indicate that [11C]forskolin would be a useful imaging agent for the AC-related second messenger system. PMID- 8229240 TI - In vitro uptake of technetium-99m-teboroxime in carcinoma cell lines and normal cells: comparison with technetium-99m-sestamibi and thallium-201. AB - Since 201Tl, 99mTc-sestamibi and 99mTc-teboroxime concentrate in cardiac cells through different mechanisms, we compared their uptake in cultured normal cells and carcinoma cell lines in order to define their possible use for tumor evaluation in vivo. Four lines of normal cells from animals, including myocytes from newborn rats, and four lines of human carcinoma cell lines were incubated for 1 hr with 37 kBq of either tracer. Results, expressed in percent of the total activity taken up by 1 million cells, showed a 9% difference between the uptake of teboroxime by normal and carcinoma lines (24.6% +/- 2.8% versus 22.5% +/- 2.1%, respectively, p < 0.05). Mean uptake was 80% higher in tumor than in normal cells for 201Tl (5.39% +/- 1.33% versus 3.00% +/- 1.08%, respectively, p < 0.001) and nearly 4 times higher for sestamibi (5.37% +/- 2.34% versus 1.44% +/- 1.88%, p < 0.001). For both agents, uptake by the myocytes and carcinoma cells was comparable (5.14% +/- 0.11% for 201Tl and 5.28% +/- 1.03% for sestamibi). When the myocytes are excluded from the group of normal cells, the uptake is 112% higher in tumor than in normal cells for 201Tl (5.39% +/- 1.33% versus 2.54% +/- 0.44%, p < 0.001) but it becomes nearly nine times higher for sestamibi (5.37% +/ 2.34% versus 0.60% +/- 0.23%, p < 0.001). It is concluded that these experiments show that the uptake of sestamibi was the most discriminant to separate between normal and malignant cells, while teboroxime was the less discriminant. Potential clinical applications for tumor visualization based on differences in sestamibi and teboroxime uptake could be envisioned. PMID- 8229241 TI - Labeling of monoclonal antibodies with rhenium-186 using the MAG3 chelate for radioimmunotherapy of cancer: a technical protocol. AB - A detailed technical protocol is provided for reproducible and aseptical production of stable 186Re-monoclonal antibody conjugates. Labeled Mab E48 IgG and its F(ab')2 fragment which are promising candidates for radioimmunotherapy of squamous cell carcinoma of the head and neck were used for evaluation. S benzoylmercaptoacetyltriglycine (S-benzoyl-MAG3) was used as a precursor. Rhenium 186-MAG3 was prepared via a unique solid-phase synthesis, after which known strategies for esterification and conjugation to Mab IgG/F(ab')2 were applied. The biodistribution of 186Re-E48 F(ab')2 in tumor-bearing nude mice was found to be comparable to that of analogously labeled 99mTc-E48 F(ab')2 or 131I-E48 F(ab')2, indicating that the intrinsic behavior of the antibody remains preserved when using this labeling technique. Radiolytic decomposition of 186Re-E48 IgG/F(ab')2 solutions of 10 mCi.ml-1 was effectively reduced by the antioxidant ascorbic acid. Upon increase of the Re-MAG3 molar amount, a conjugation of seven to eight Re-MAG3 molecules per Mab molecule was generally the maximum ratio that could chemically be obtained. Such a ratio did not impair the immunoreactivity or alter the in vivo biodistribution characteristics of the immunoconjugate, making this labeling procedure suitable for general clinical application. PMID- 8229242 TI - Technetium-99m-labeled hydrazino nicotinamide derivatized chemotactic peptide analogs for imaging focal sites of bacterial infection. AB - We synthesized and evaluated four hydrazino nicotinamide (HYNIC) derivatized chemotactic peptide analogs: For-NleLFK-HYNIC (HP1), For-MLFK-HYNIC (HP2), For MLFNH(CH2)6NH-HYNIC (HP3), and For-MLF-(D)-K-HYNIC (HP4), for in vitro bioactivity and receptor binding. The peptides were radiolabeled with 99mTc via a glucoheptonate co-ligand and their biodistribution determined in rats (n = 6/time point) at 5, 30, 60 and 120 min after injection. Localization of the peptides at sites of deep thigh Escherichia coli infection was determined by radioactivity measurements on excised tissues in rats (n = 6/time point) and rabbits as well as scintillation camera imaging in rabbits (n = 6). All peptides maintained biological activity (EC50s for O2 production by human PMNs: 12-500 nM) and the ability to bind to the oligopeptide chemoattractant receptor on human PMNs (EC50s for binding: 0.12-40 nM). After incubation with 99mTc-glucoheptonate, radiolabeled peptides were isolated by HPLC at specific activities of > 10,000 mCi/microM. Technetium-99m-labeled peptides retained receptor binding with EC50s < 10 nM. Blood clearance of all four peptides was rapid. Biodistributions of the individual peptides were similar, with low levels of accumulation in most normal tissues. In rats, all of the peptides concentrated at the infection sites (T/B ratio: 2.5-3:1) within 1 hr of injection. In rabbits, outstanding images of the infection sites were obtained, with T/B ratios of > 20:1 at 15 hr after injection. This study demonstrates that 99mTc-labeled chemotactic peptide analogs are effective agents for the external imaging of focal sites of infection. PMID- 8229243 TI - Localization of indium-111-immunoglobulin G, technetium-99m-immunoglobulin G and indium-111-labeled white blood cells at sites of acute bacterial infection in rabbits. AB - Biodistribution and infection imaging properties of 111In-DTPA-IgG, 99mTc hydrazino nicotinamide-IgG and 111In-WBC were compared in rabbits with E. coli infection. Groups of six rabbits were injected with 10 mCi of 99mTc-IgG plus 0.5 mCi of 111In-IgG or 1 mCi of 99mTc-IgG plus 0.05 mCi of 111In-WBC. At 4-5 and 18 20 hr, dual photon scintigrams were acquired. At both times, the distributions of 99mTc and 111In-IgG were nearly identical. The sites of infection were well visualized with all three radiopharmaceuticals. In the early images, the target to-background ratios (T/B) for 111In and 99mTc-IgG determined by ROI analysis were 1.95 +/- 0.26 and 2.57 +/- 0.38 (p = NS). In the delayed images, the T/B ratios increased (p < 0.01) to 3.56 +/- 0.49 and 4.90 +/- 0.98. At both times, the T/B ratios for 111In-WBC were higher (p < 0.01); 4.17 +/- 0.78 at 4-5 hr and 8.52 +/- 1.52 at 18-20 hr. These results indicate that all three agents yield excellent images of infection sites. Although 111In-WBC had higher T/B ratios, the ease of preparation of the radiolabeled proteins makes them attractive alternatives for infection imaging. PMID- 8229244 TI - Lobar primary pulmonary lymphoma: iodine-123-iodoamphetamine and thallium-201 chloride scintigraphic findings. AB - A case of primary pulmonary malignant lymphoma (PML) was evaluated by 123I-IMP scintigraphy and 201TlCl SPECT. Radiography and CT showed a so-called lobar consolidation on the chest that was difficult to differentiate from other benign conditions. However, 123I-IMP scintigraphy showed a defect corresponding to the lesion both in the early and delayed images, and 201TlCl SPECT showed a higher retention index than that of benign disorders. Both results were highly suggestive of malignancy. These scintigraphies can help differentiate PML from benign lesions and other malignant pulmonary tumors. PMID- 8229245 TI - Cerebral vasodilatory capacity mapping using technetium-99m-DTPA-HSA SPECT and acetazolamide in moyamoya disease. AB - A cerebral vasodilatory capacity map, consisting of a 99mTc-DTPA-HSA SPECT image obtained after acetazolamide injection minus the baseline image, was produced in a patient with moyamoya disease. The map demonstrated diminished capacity in the posterior region of the right cerebral hemisphere. Subsequently, cerebral infarction occurred in the corresponding area. This observation suggests that regions of low vasodilatory capacity identified by cerebral vasodilatory capacity mapping may be at high risk of ischemic stroke. PMID- 8229246 TI - Increased cerebral perfusion after detachable balloon embolization of carotid cavernous fistula on technetium-99m-HMPAO brain SPECT. AB - Most symptoms and signs associated with a carotid cavernous fistula (CCF) are thought to be related to regurgitation of flow into cortical veins and to venous congestion. Arteriovenous shunting and secondary perfusion insufficiency is regarded as less important in causing symptoms. We describe a 27-yr-old male patient who had improvement of neurologic symptoms and signs after detachable balloon embolization of traumatic CCF. The pre- and postocclusion 99mTc-HMPAO brain SPECT scan showed improved cerebral blood flow after occlusion. The CCF had shown marked arteriovenous shunting without significant venous congestion on pre occlusion cerebral angiogram. The postocclusion cerebral angiogram revealed complete occlusion of the CCF with increased blood flow in the ipsilateral middle cerebral artery distribution. These findings suggest that cerebral dysfunction may be related to perfusion insufficiency from the CCF. Brain SPECT scanning can assess the functional status of cerebral perfusion and may be a useful, noninvasive adjunct to angiography. PMID- 8229247 TI - Osteosarcoma presenting as intestinal obstruction. AB - Osteosarcoma commonly presents with osseous and pulmonary metastases. We present an unusual case of extraosseous metastatic abdominal chondroblastic osteosarcoma presenting as intestinal obstruction. PMID- 8229248 TI - Pancoast tumor as a cause of reflex sympathetic dystrophy. AB - Reflex sympathetic dystrophy of the upper extremity can be triggered by a wide variety of factors. Pancoast tumor should be added to the list of precipitating conditions which can induce this syndrome. PMID- 8229249 TI - Quantitative perfusion imaging assessing acquired discrete peripheral pulmonary artery stenosis. AB - We present the case an 8-yr-old boy evaluated for anastomotic stenosis of the right pulmonary artery after surgical repair of hemitruncus at 6 wk of age. Pulmonary angiography revealed only mild narrowing and a 10-mm pressure gradient across the anastomosis, but quantitative perfusion imaging demonstrated that the right lung only received 16% of pulmonary blood flow. Subsequently, balloon angioplasty of the anastomotic site was performed, resulting in complete resolution of the stenosis and gradient. Early postangioplasty perfusion imaging demonstrated increased perfusion of the right lung to 35% of total pulmonary blood flow. It is theorized that initially a chronically hyperperfused lung may develop more capacious vessels and recruit new capillaries during the years of hyperperfusion such that a "perfect" angioplasty may result in less than symmetric perfusion. The inexpensive, noninvasive quantitative perfusion study is more sensitive and accurate in evaluating acquired (postsurgical) pulmonary artery stenoses. PMID- 8229250 TI - Cardiac phantom evaluation of simultaneously acquired dual-isotope rest thallium 201/stress technetium-99m SPECT images. AB - Simultaneously acquired dual-isotope 201Tl/99mTc SPECT studies were performed using cardiac and thoracic phantoms to evaluate the dual-isotope myocardial perfusion technique. Cardiac phantom images representing infarction, viable myocardium and various levels of ischemia were analyzed. Studies with and without attenuating media were performed, and myocardium-to-defect count ratios and defect sizes from dual-isotope SPECT images were compared to myocardium-to-defect count ratios and defect sizes from single-isotope (201Tl and 99mTc) SPECT images. Dual-isotope studies also were interpreted qualitatively. Studies with background activity simulating clinical conditions were performed and interpreted qualitatively. Myocardium-to-defect count ratios from both 99mTc and 201Tl were similar in single-isotope and dual-isotope SPECT images. Thallium-201 and 99mTc defect sizes were decreased slightly (mean +/- s.d., 1.0 +/- 1.7 cc for 201Tl and 0.7 +/- 1.0 cc for 99mTc) on dual studies when compared to single studies but were not statistically significant. Dual-isotope image simulations of normal, ischemic and infarcted and viable myocardium were correctly identified by experienced clinicians in 95% of the cases (21/22). Simultaneous dual-isotope 201Tl/99mTc SPECT imaging of cardiac phantoms produced images that had similar myocardium-to-defect count ratios to those produced using single-isotope techniques and were correctly evaluated on qualitative analysis. Changes in defect size related to dual-isotope imaging were minimal and not qualitatively important. PMID- 8229251 TI - Simultaneous thallium-201/technetium-99m dual-isotope cardiac SPECT: ready for prime time? PMID- 8229252 TI - Automated iterative three-dimensional registration of positron emission tomography images. AB - Two types of image similarity measures, the sum of absolute differences (SAD) and the stochastic sign change (SSC), were compared for three-dimensional registration of images from PET. To test the accuracy of both registration methods, 30 FDG brain studies, 40 13N-ammonia cardiac studies and 20 FDG liver tumor studies (where each image set contained 15 image planes, 128 x 128 pixels per plane) were made into worse case conditions by creating image sets of low counts and extreme defects. These images were then registered to the reference images that had been moved in three dimensions into a random set of known translations, rotations and normalization factors (x, y, z, theta, rho, sigma, nf). Neither method required any external fiduciary markers or operator interventions to register a set of images. The optimization of the image similarity (using the SAD or SSC) was performed with the simplex method and registration was completed within 10 min of computation time on a low-end workstation. Overall, the SAD method had an average inplane (x, y) registration error of 0.5 +/- 0.5 mm, a z-axis registration error of 1.1 +/- 1.1 mm, an inplane rotational error of 0.5 +/- 0.4 degrees, an out-of-plane rotational error of 1.1 +/- 1.2 degrees and a normalization factor error of 0.015 +/- 0.016. The SSC method had an average inplane (x, y) registration error of 0.6 +/- 0.5 mm, a z-axis registration error of 1.1 +/- 1.1 mm, an inplane rotational error of 0.7 +/- 0.5 degrees, an out-of-plane rotational error of 1.0 +/- 1.2 degrees and a normalization factor error of 0.014 +/- 0.014. This study demonstrates that either the SAD or SSC method for measuring image similarity, combined with the simplex method for function optimization, are accurate methods for registration of a wide variety of PET images including low count studies and those with marked interval changes in the pattern of count distribution. PMID- 8229253 TI - Principal axes and surface fitting methods for three-dimensional image registration. AB - We evaluated the effect of the image acquisition parameters on the accuracy of the principal axes and surface-fitting techniques for three-dimensional image registration. Using two types of phantom objects, MR brain image and a mathematically defined ellipsoid, we simulated pairs of scans with known acquisition parameters, including longitudinal coverage, magnitude of mis registration, number of sections and section thickness. Both methods are sensitive to the systematic deformation of contours. The principal axes method is also sensitive to incomplete scan coverage and to the x-axis and y-axis misangulation. Both methods are insensitive to the number of sections, section thickness and the number of points per section. Surface fitting performed well without user supervision. There is no need for routine inclusion of the scaling factors as search parameters. The results confirm the feasibility of three dimensional multimodality registration of brain scans with accuracy 1-2 mm, with surface fitting being the method of choice. PMID- 8229254 TI - Hepatobiliary handling of iodine-125-Tyr3-octreotide and indium-111-DTPA-D-Phe1 octreotide by isolated perfused rat liver. AB - Radiolabeled bioactive peptides may show receptor-mediated binding to tumors, making them suitable for scintigraphic imaging. The liver is an important organ for peptide clearance. To gain insight into the uptake and intracellular processing of somatostatin analogs, we compared the hepatobiliary handling of 125I-Tyr3-octreotide and 111In-DTPA-D-Phe1-octreotide, which are successfully used to image somatostatin receptor-positive tumors in vivo in isolated recirculating perfused rat livers. Sixty minutes following administration of the radiolabeled peptides, perfusion medium and biliary radioactivity were analyzed. Radioiodinated Tyr3-octreotide was rapidly cleared by the liver and 60% of the dose was excreted intact into the bile after 60 min. In contrast, 111In-DTPA-D Phe1-octreotide was not cleared by the liver; medium radioactivity levels remained about constant and only 2% of the dose was found in the bile. These results are in agreement with in vivo findings in rats and humans. We concluded that isolated rat liver perfusion is a good system to rapidly gain insight into the hepatic handling of radiopharmaceuticals. PMID- 8229255 TI - Chromatographic identification in serum of endogenously radioiodinated thyroid hormones after iodine-131 whole-body scintigraphy in the follow-up of patients with differentiated thyroid carcinoma. AB - Patients with differentiated thyroid cancer (DTC) are conventionally followed with serial 131I whole-body scintigraphy (WBS) and serum thyroglobulin (hTg) assay. Given the 15%-20% incidence of discordant results, we developed a sensitive and specific procedure for monitoring such patients, based on the assumption that 131I uptake, even if too low to be detected by 131I WBS, could be assayed in serum as thyroid products (hTg, T3 and T4) endogenously labeled with 131I. Our study included 125 patients routinely monitored for tumor recurrence or for the persistence of functioning thyroid tissue after complete primary treatment for DTC (surgery and 131I ablation of remnants). A plasma sample, taken 72 hr after administering 131I for WBS was fractionated on a Sephadex-G25 superfine column by first eluting all of the radioactive species except the thyroid hormones and then the radioiodothyronines. The sensitivity and specificity of chromatography in detecting functioning thyroid tissue after primary treatment for DTC were 98.4% and 100% (accuracy 99.2%), respectively, versus 90.6% and 95.1% for 131I WBS (accuracy 92.8%) and 60.9% and 100% for hTg (accuracy 80%). Combining chromatography with serum hTg gave the highest gains in diagnostic performance (100% for all parameters). This chromatographic method can be used in addition to conventional procedures in the follow-up of patients with DTC and represents a highly sensitive test for assessing the results of 131I ablation of postsurgical remnants. PMID- 8229256 TI - A refined method for quantification of myocardial oxygen consumption rate using mean transit time with carbon-11-acetate and dynamic PET. AB - The utility of the mean transit time equation was investigated for estimation of the myocardial clearance rate constant of 11C-acetate, which is proportional to myocardial oxygen consumption rates. The mean transit time approach was also employed to generate parametric images of the clearance rate constant of 11C acetate with dynamic PET imaging in 20 normal human studies. Input function delays and cutoff errors due to the truncation of the myocardial tissue time activity curve at a finite time were corrected. The clearance rate constants estimated by mean transit time correlated well with the estimates by conventional monoexponential fitting (15 min (truncation time): Y = 0.01 + 0.94X, correlated coefficient (r) = 0.99; 16 min: Y = 0.03 + 0.94X, r = 0.98; 20 min: Y = 0.03 + 0.84X, r = 0.99). The clearance rate constants estimated by the mean transit time approach also correlated well (r = 0.94) with the measured rate-pressure products. The quality and noise level of parametric images of the clearance rate constants generated by mean transit time are improved over those generated by monoexponential fitting. Additional advantages of the mean transit time approach compared to the standard monoexponential fitting method for estimating myocardial clearance rate constant of 11C-acetate include ease of input function delay correction, less sensitivity to the shape of the input function and elimination of subjective data selection of the linear portion of the clearance data on a semilog plot. Thus, this approach is expected to facilitate objective quantitative analysis of indices of myocardial oxygen consumption. PMID- 8229257 TI - Quantitative brain SPECT in Alzheimer's disease and normal aging. AB - To improve the diagnostic utility of brain single-photon emission computed tomography (SPECT) in Alzheimer's disease (AD), we have developed and evaluated an objective method of differentiating patients and healthy elderly controls using a quantitative image analysis protocol. HMPAO-SPECT image datasets from 29 patients with probable AD and 78 age-matched controls were registered to a common anatomic frame of reference. Activity levels within 120 standardized cortical volumes were determined by an automated procedure. Subjects were classified into normal and AD groups by quadratic discriminant analysis using two features: global average activity level and average normalized activity levels within the two clusters of standardized volumes identified as most significantly different in AD by analysis of covariance. The classification used split-half replication to ensure valid results. Classification performance quantified by the area under a binormal ROC curve fitted to the data was 0.923 +/- 0.036; at a threshold likelihood ratio of 1, the sample sensitivity was 91% and specificity was 86%. We conclude that quantitative SPECT accurately distinguishes AD patients from elderly controls. PMID- 8229258 TI - Noninvasive diagnosis of cardiac transplant arteriopathy with dipyridamole thallium scintigraphy. PMID- 8229259 TI - Nuclear medicine and health care reform. PMID- 8229260 TI - Critical thinking and clinical judgment of professional nurses in a career mobility program. AB - This study's purpose was to establish the impact of a Career Advancement Program on critical thinking and clinical judgment of Registered Nurses. Findings of the study suggest that critical thinking and clinical judgment are not influenced by patterns of upward mobility. Additionally, an analysis did not indicate significant predictors in education, specialty area, or years of nursing experience related to scores. Whereas this study questions the efficacy of career mobility programs on critical thinking and clinical judgment, it does not measure the impact of career advancement programs on other key outcome variables. The study suggests nurse educators emphasize critical thinking and clinical judgment rather than transmission of information in educational offerings. PMID- 8229261 TI - A survey of the qualifications, special training, and levels of personnel working emergency department triage. AB - Emergency department triage is a challenging position for the Registered Nurse, and one that requires special qualifications, training, and skills to adequately assess, prioritize, and intervene in a variety of client presentations. This survey, conducted throughout the United States in 1991, reports the varied qualifications and special training required by hospitals for nurses to work triage. The categories of personnel who work triage are also reported. The reasons for the variability and diversity in qualifications and training between geographical zones are discussed, along with some creative suggestions for providing additional education and training. PMID- 8229262 TI - The role of the nurse educator in the development of critical pathways. AB - This article focuses on the role of the nurse educator in developing critical pathways. It serves as a guide for the nurse educator by defining how critical pathways support managed care, identifying the goals of critical pathway education, and reviewing the importance of organizational commitment to the critical pathways before their use. The nurse educator serves as instructor, facilitator, and consultant in critical pathway development. PMID- 8229263 TI - A two-level approach to nursing process. AB - Nurses have problems integrating the nursing process and care plans into their nursing practice. This problem affects patient outcomes and nursing practice issues. Encouraging trends have been observed when nurses adopted a two-level approach to relearn and implement the nursing process. Assessments were more comprehensive, nursing diagnoses were more specific, precise, and accurate, documentation was improved, and care plans were individualized. This article offers step by step guidelines to learn the two-level approach while recommending methods for teaching. PMID- 8229264 TI - Decentralization of the staff development department in home care. AB - An easy transition to home care and improved quality of orientation are important factors for home care agencies considering decentralization of the staff development department. A Visiting Nurse Association found that the positive advantages of decentralization outweighed the initial adjustment problems. PMID- 8229265 TI - Taking the fear out of the JCAHO Survey. PMID- 8229266 TI - Communicating float staff competency validation. PMID- 8229267 TI - Personal information managers. Is there a PIM in your future? PMID- 8229268 TI - Development needs of advance practice nurses in a managed care environment. AB - The authors describe an organizational change process driven by a critical cost containment effort in a teaching hospital in the northeastern United States. This process resulted in a major shift in the role of the clinical nurse specialist. Development priorities of clinical nurse specialists before and after redesign of the role are described. PMID- 8229269 TI - The hospital-Visiting Nurse Association partnership. A continuous quality improvement program. AB - The authors present a model for a joint continuous quality improvement program between a home health agency and a referring hospital. The program evaluates the patient's healthcare continuum and provides feedback to both organizations regarding the effectiveness of discharge planning, referral, and home care services. A program such as this, which fosters the sharing of information and collaboration on common goals, is essential to care coordination. Information criteria used to design the model, methodologies for implementing evaluation activities, and mutual benefits realized by the participating organizations are described. PMID- 8229270 TI - Changing the delivery of nursing care. Implementation issues and qualitative findings. AB - Faced with a shortage of professional nurses, 37 hospitals in New Jersey implemented a variety of approaches to changing the delivery of nursing care. Implementation issues, including the positive and negative effects of the changes, are discussed. Most project coordinators reported that implementation was more difficult than they had anticipated. PMID- 8229271 TI - Relationship of family satisfaction to employee job satisfaction. PMID- 8229272 TI - Planning and program development for psychiatric home care. AB - The psychiatric home care program of the Visiting Nurse Association and Hospice of Northern California (VNAHNC) was developed in March of 1984. The largest such program in the Bay area, the program generated a revenue of more than $1 million, made 9,756 visits, and served more than 500 patients in 1992. When discussing the program at two national conferences, members of the home care community asked many questions about the feasibility of starting up their own psychiatric home care programs. This article addresses the most frequently asked questions. PMID- 8229273 TI - Administrative delays. Is there a difference between for-profit and nonprofit hospitals? AB - Unnecessary administrative delays can account for significant costs if not identified and managed properly. This study compares the types of delays, the length of stay, and the estimated costs at a for-profit hospital and a nonprofit hospital. There was no significant difference between the two hospitals in terms of length of stay. A significant difference in cost was found, however. The major delay problems were related to unavailable outside resources, discharge planning, and physician involvement. PMID- 8229274 TI - A managed care system. Financial and clinical evaluation. AB - The ever-changing healthcare environment requires that nurse executives explore alternative ways to deliver quality patient care, manage resources, and reduce length of stay. Care paths provide one such alternative, incorporating multidisciplinary care planning, discharge planning, and documentation in a patient-centered design. This model includes a clinical and financial system for evaluating the impact of the care path on resource utilization, length of stay, and patient outcomes. PMID- 8229275 TI - Identifying nursing research priorities in an acute care hospital. AB - The purpose of this study was to identify nursing research priorities for an acute care hospital. Using the Delphi technique, data were obtained from a panel of 52 nurses who held administrative, staff, or support positions. After three rounds of data collection, a prioritized listing of potential research topics was obtained. The study results have helped motivate and guide research efforts, stimulate formation of research interest groups, and attract local researchers to the facility. PMID- 8229276 TI - A study of hospital based nursing case management. PMID- 8229277 TI - A collaborative approach to home OKT3 infusion therapy. AB - Hospitalization of patients who have undergone renal transplantation is often extended because of the need to complete complex antirejection infusion therapy. Because of the costs associated with prolonged hospitalization, medical professionals who provide care to these patients are developing programs that will facilitate attainment of quality care in the outpatient setting. The authors describe the effective implementation of a home Muromonab-CD3 (OKT3) intravenous administration program among renal transplant recipients based on thorough planning and close collaboration between the transplant service and home care staff. PMID- 8229278 TI - Nursing leadership and healthcare reform. Part II: Nursing practice in a reformed healthcare system. PMID- 8229279 TI - Nursing education and violence against women. PMID- 8229280 TI - Student clinical experiences in shelters for battered women. AB - This article describes the critical social problem of wife battering and clinical experiences for student nurses in shelters for battered women. The argument is presented that due to increasing frequency and severity, nurses must become more aware of and effective in caring for battered women and their children. In keeping with the call from nursing leaders and health care reformers, the shelter clinical experience provides students with an opportunity to shift from a focus on high technology to community-based primary care. Students are able to function as advocates, advisers, case managers, and health care teachers, and to focus on broad concerns of daily living problems. Lindeman's four recommendations for teaching in clinical settings were used as a framework for this article. PMID- 8229281 TI - A model for the integration of HIV/AIDS content into baccalaureate nursing curricula. AB - The HIV/AIDS epidemic has posed unprecedented challenges for nurse faculty. The purpose of this article is to describe a model for the integration of HIV/AIDS content into health professions' curricula over time, specifically into baccalaureate nursing programs. The model was developed through nurse faculty experience with a professional AIDS education project over five years, with over 16,000 trainees. The model includes three stages: the imported guest speaker, mainstreaming, and integration. In the last stage, it is noted that HIV/AIDS serves as a vehicle for curricular development that prepares nurses to meet care problems beyond those limited to HIV. It is hoped that this model will be useful for nurse educators now facing the complexities of curriculum development necessary in response to HIV/AIDS. PMID- 8229282 TI - Community health learning experiences and political activism: a model for baccalaureate curriculum revolution content. AB - Nursing education is involved in a curriculum revolution that decries behaviorist theory. One model for the new educational approaches is political activism that involves faculty and students in experiences of caring client advocacy. Content, theory, and experiences in community-health nursing integrate community activism and sociopolitical concepts. Sample approaches that blend curriculum revolution and political activism concepts are included. Such approaches can easily be used as part of the lived experience of community-health nursing education. PMID- 8229283 TI - Effect of a program to increase the cognitive level of questions asked in clinical postconferences. AB - This study examined the effect of a program to increase the percentage of cognitively high-level questions asked in clinical postconference. Fourteen faculty and the students in their clinical groups comprised the convenience sample. Pre- and postintervention data on the cognitive level of questions asked by the treatment (N = 10) and control (N = 4) groups were collected by way of audiotapes. Treatment group faculty participated in an intervention designed to improve their use of questions. Prior to the intervention, there was no statistically significant difference in the cognitive level of questions asked by faculty or students in the treatment and control groups. Following the intervention, faculty in the treatment group asked a statistically significant higher percentage of cognitively high-level questions (p = 0.012 [one-tailed]), but there was no statistically significant difference between the students in the treatment and control groups. PMID- 8229284 TI - The teacher as midwife: a New Zealand narrative. PMID- 8229285 TI - A study of professional health care students' attitudes toward older adults. PMID- 8229286 TI - Teaching novice to expert: the transformed teacher-student relationship. PMID- 8229287 TI - Community participation in curriculum development: a three-pronged approach. PMID- 8229288 TI - The Honors Program: pathway to excellence. PMID- 8229289 TI - Greening the curriculum. PMID- 8229290 TI - Amino acids suppress intracellular protein degradation in rat liver during parenteral nutrition. AB - The effects of variations in the amino acid supply on the rates of synthesis and degradation of liver proteins and on the rate of synthesis and secretion of plasma proteins were investigated. Rats were nourished by infusion of total parenteral nutrition solutions containing four different levels (0, 1.65, 3.3 and 6.6%) of amino acids for 7 d. The fractional rate of total protein synthesis in the liver was determined by injecting a flooding dose of [3H]phenylalanine. The proportion of newly synthesized proteins retained and exported by the liver was estimated by injecting a tracer dose of [14C]leucine and then measuring the protein radioactivity remaining in the liver and present in the plasma after secretion was completed. The rate of plasma albumin synthesis was significantly lower in the 0 and 1.65% amino acid groups than in the other groups. The fractional synthesis rates of liver domestic proteins, however, were essentially the same in rats administered all levels of amino acids except for the 0% amino acid group, which showed a slightly higher value than the other groups. The fractional degradation rates of liver domestic proteins, calculated as the difference between the fractional synthesis rate of liver domestic proteins and the net gain of liver proteins, were found to be inversely related (r = -0.999, P < 0.05) to the level of amino acids in infusion solutions up to 3.3% amino acids. It was concluded that protein degradation plays the predominant role in the regulation of liver protein mass. PMID- 8229291 TI - 1,25-dihydroxycholecalciferol analogs cannot replace vitamin D in normocalcemic male rats. AB - A possible role of vitamin D in the growth and development of rats was investigated. Impaired development was observed in normocalcemic, vitamin D deficient male and female rats, as revealed by low intestinal calcium transport, low renal vitamin D receptor levels and poor bone mineralization. Analogs of 1,25 dihydroxycholecalciferol, possessing reduced calcium-mobilizing activity in intestine and bone but retaining differentiation activity in cultured cells, were unable to support normal development of normocalcemic, vitamin D-deficient male rats. These results suggest that either the calcium-mobilizing activity alone or both the calcium-mobilizing activity and differentiating activity of vitamin D are required for normal development or that the analogs are inactive in vivo. We also demonstrated sex-related differences in intestinal calcium transport, renal vitamin D receptor regulation and bone mineralization that were independent of vitamin D status. PMID- 8229292 TI - Dietary aluminum chloride reduces the amount of intestinal calbindin D-28K in chicks fed low calcium or low phosphorus diets. AB - The mechanism of aluminum (Al) toxicity may involve disturbances in calcium (Ca) metabolism. Aluminum compounds have been reported to reduce vitamin D-dependent Ca absorption in chicks, rats and humans. To investigate the mechanism by which Al reduces Ca absorption, we studied the effect of dietary aluminum chloride (AlCl3) on the relative amounts of intestinal calbindin D-28K in chicks fed diets varying in Ca and phosphorus concentration. AlCl3 was added so that Al constituted 0, 0.15 or 0.3 g/100 g of diets that were either adequate, low in Ca, low in P, or contained supplemental P. Diets were fed for 2 wk. Intestinal calbindin D-28K levels were assayed using SDS-PAGE and 45Ca binding to Western blots. Added dietary Al greatly reduced the amount of intestinal calbindin in chicks fed adequate diets, low Ca diets, or low P diets. When diets with supplemental P were fed, little calbindin was evident with or without added Al. Tibia ash, body weight and food intake were also reduced (P < 0.05) by added Al. These results indicate that dietary AlCl3 inhibits vitamin D-dependent Ca absorption by reducing the amount of intestinal calbindin D-28K. Aluminum, therefore, may interfere with the body's ability to regulate intestinal calbindin D-28K levels. This could have implications for other tissues that contain substantial levels of calbindin D-28K. PMID- 8229293 TI - Isolated hearts from copper-deficient rats exhibit improved postischemic contractile performance. AB - Dietary copper deficiency affects a number of enzymes, the function of which may influence the outcome of myocardial ischemia-reperfusion injury. Male weanling rats were fed diets that were adequate (> 5 mg/kg) or deficient (< 1 mg/kg) in copper. After 4 wk, the rats' hearts were isolated and used to study the effects of ischemia-reperfusion on intraventricular developed pressure (DevP), positive and negative rates of intraventricular pressure change (+dp/dt and -dp/dt) and release of lactate dehydrogenase and creatine kinase from the heart. The ischemia perfusion protocol included a 15-min equilibration period, 30 min of warm, total ischemia and reperfusion for 30 min. Preischemic hearts from copper-deficient rats produced lower DevP than hearts from copper-adequate rats at all levels of preload. However, postischemic recovery of DevP was significantly greater in the hearts of the copper-deficient group. Furthermore, the postischemic patterns of lactate dehydrogenase and creatine kinase release in the two groups were significantly different. These findings indicate that, although dietary copper deficiency adversely affects a number of enzymatic systems, the functional recovery of hearts subjected to ischemia-reperfusion injury is improved when the diet is restricted in copper. PMID- 8229294 TI - Hydroxycobalamin[c-lactam] increases total coenzyme A content in primary culture hepatocytes by accelerating coenzyme A biosynthesis secondary to Acyl-CoA accumulation. AB - Hydroxycobalamin[c-lactam] (HCCL) treatment in rats results in decreased hepatic L-methylmalonyl-CoA mutase activity and increased hepatic total CoA content. To test the hypothesis that HCCL increases hepatic CoA biosynthesis secondary to propionyl- and methylmalonyl-CoA accumulation. CoA homeostasis was studied in primary culture rat hepatocytes. Conversion of [14C]pantothenic acid to [14C]CoA in the primary culture system was accelerated to rates 3-5 times control by acute incubation with the acyl-CoA-generating carboxylic acids pivalate (10 mmol/L) or propionate (10 mmol/L). HCCL (1 mg/L included from 24 to 72 h of culture) had no affect on the distribution of the hepatocyte CoA pool or total CoA content. However, culture in the presence of HCCL and propionate (2 mmol/L) resulted in accumulation of methylmalonyl-CoA and a 59% increase in total CoA content. The combination of HCCL (1 mg/L) and propionate (0.5 mmol/L), but not HCCL alone, increased the rate of [14C]pantothenic acid conversion to [14C]CoA by 150%. Degradation of [14C]CoA in the primary culture hepatocyte system was unaffected by chronic exposure to 2 mmol/L propionate, but was increased 90% by the combination of HCCL and 2 mmol/L propionate. Thus, in the presence of a source of propionyl-CoA, HCCL treatment results in methylmalonyl-CoA accumulation and accelerated CoA biosynthesis. The increased CoA biosynthesis leads to increased hepatocyte total CoA content, which may contribute to cellular metabolic homeostasis under conditions of acyl-CoA accretion. PMID- 8229295 TI - Dietary fat and fiber alter rat colonic mucosal lipid mediators and cell proliferation. AB - To better understand the biochemical mechanisms by which dietary fat and fiber modulate colonic cell proliferation, we determined the effect of dietary fats and fibers on rat colonic epithelial cell phospholipid mass and composition and on two metabolic products of phospholipids, prostaglandins and diacylglycerol (DAG). In a 3 x 3 factorial design, groups of 10 male Sprague-Dawley rats were fed one of nine experimental diets for 3 wk: three types of fat at 15 g/100 g (beef tallow, corn oil or fish oil) x two types of fiber (pectin or cellulose) or fiber free as a control group. Dietary treatment did not alter phospholipid or DAG mass, although the fatty acid compositions of membrane phospholipids and DAG were altered by dietary treatment. Arachidonic acid [20:4(n-6)] and eicosapentaenoic acid [20:5(n-3)] in colonic mucosal phospholipid and DAG were associated with higher and lower indices of cell proliferation, respectively. These correlations were specific for the distal colon, which was the principle site of dietary fat effects on cell proliferation. Prostaglandin E and prostacyclin synthesis in colonic mucosa and muscle was significantly lower in fish oil-fed compared with beef tallow- and corn oil-fed animals (by 46-90%, P < 0.001), in both the proximal and distal colon. Correlations between prostaglandin production and cell proliferation, however, were significant only in the distal colon. These data raise the possibility that dietary fat and fiber may modulate intracellular events related to cell proliferation via their effects on epithelial cell phospholipid fatty acid composition, and subsequently on prostaglandin production and DAG composition. PMID- 8229296 TI - Protein restriction during pregnancy affects postnatal growth in swine progeny. AB - Protein deficiency during pregnancy affects fetal development. The critical period, when the fetus is most susceptible to maternal protein deficiency and its effect on neonatal growth, is unknown. Therefore, we studied the effect of a protein-restricted diet during early and late pregnancy and throughout pregnancy on growth of pigs from birth to market weight. Sows were fed a control (13% protein) or protein-restricted (0.5% protein) diet throughout pregnancy or protein-restricted diet from d 1 to 44, then control diet to term or control diet from d 1 to 81, then the protein-restricted diet to term. In Experiment 1, birth weights were measured, and 12 pigs/diet group were weaned at 4 wk and raised to market weight. Feeding the protein-restricted diet throughout pregnancy reduced birth and slaughter weights, whereas the control followed by protein-restricted and protein-restricted followed by control diets reduced only birth weight relative to controls. Indices of carcass lean were reduced in the protein restricted piglets, with carcass fat not affected. In Experiment 2, control and control-protein-restricted litters were reduced to six piglets and 3/litter cross fostered to a sow of the other treatment group. After weaning at 4 wk, 4 piglets/group were individually fed to 8 wk. The control and control followed by protein-restricted diet fed piglets had similar weights at birth, but piglets raised by a control-protein-restricted sow tended to weight less at weaning than their littermates raised by a control sow. After weaning, these piglets had greater feed intakes relative to other groups and there were no weight differences by 8 wk.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229297 TI - 45Ca kinetics in growing miniature poodles challenged by four different dietary levels of calcium. AB - In 21 miniature poodles the 45Ca metabolism was studied at 13, 19 and 25 wk of age. The animals were divided in four groups and fed dry dog food differing only in Ca concentration, i.e., 0.05 (LLC), 0.33 (LC), 1.1 (NC) or 3.3 g/100 g (HC) of Ca on a dry matter basis. The percentage of Ca absorption was significantly higher in the LLC and LC groups (70-96%) than in the NC group (28-53%). True Ca absorption was low and retention almost zero in the LLC dogs, whereas in the HC dogs absorption and retention were more than three times greater than in the NC dogs. The higher retention in the HC group compared with the NC group was primarily the result of a low level of bone removal, the deposition levels being similar in the two groups. We conclude that 1) the 0.33 g/100 g of Ca diet did not meet the minimal requirements for growth in small breed dogs, 2) both low and high dietary Ca caused effects on Ca metabolism similar to those previously observed in giant breed dogs, and 3) small and giant breed dogs require different minimal Ca:energy ratios in the diet during growth. PMID- 8229298 TI - Baked rye products modify cholesterol metabolism and crypt cell proliferation rates in rats. AB - Young adult male rats were made hypercholesterolemic by feeding cholesterol (+ cholic acid). The effect of rye crispbread on hepatic and plasma cholesterol concentrations, hepatic cholesterol synthesis, small intestinal cell turnover and fecal output was investigated. Diets containing 50 and 75% rye crispbread (g dry wt) were compared with positive control diets of similar gross composition, in which the insoluble component of rye was matched with cellulose and the soluble component with guar gum. A negative control diet containing no non-starch polysaccharide was also included. Cholesterol supplementation was maintained in half the rats on each diet. Rye caused a marked increase in fecal output that was greater than that seen in the positive control groups. However, crypt cell proliferation in the small intestine was less than that seen in the high fiber control groups. Rye reduced total plasma cholesterol concentrations only in those rats that continued to receive the high cholesterol diet. However, whether or not cholesterol was fed, the presence of nonstarch polysaccharide caused a decrease in liver cholesterol concentrations. Rye caused a marked increase in hepatic cholesterol synthesis over both the positive controls and the rats fed a fiber free diet. This implies that rye causes a loss of cholesterol from the body, probably due to malabsorption of bile acids and cholesterol. PMID- 8229299 TI - Biotin and biotin analogs in human urine: biotin accounts for only half of the total. AB - In studies using avidin-binding assays to measure the urinary excretion of biotin, biotin is sometimes assumed to be equal to the detectable avidin-binding substances present. High performance liquid chromatography was used to separate avidin-binding substances in human urine, and the chromatographic fractions were assayed for avidin-binding substances (biotin and biotin analogs) by a sensitive, specific assay based on binding of biotin to [125I]avidin. In a study of ten normal adults, substantial amounts of avidin-binding substances other than biotin were detected, two of which were bisnorbiotin and biotin sulfoxide. These biotin analogs were initially identified by their chromatographic properties, and identities were confirmed by chemical conversion. The presence of avidin-binding substances in addition to biotin may have confounded previous measurements of the urinary excretion of biotin using avidin-binding assays. Because bioassay methods for biotin often use organisms for which one or more of these biotin analogs are growth factors, measurements of biotin in urine using some bioassay methods are likely to overestimate the concentrations of biotin. PMID- 8229300 TI - Coconut oil and sesame oil affect lymphatic absorption of cholesterol and fatty acids in rats. AB - Five groups of male Wistar rats weighing approximately 200 g consumed 12 or 24% sesame oil or coconut oil diets or a control diet (14% corn oil) ad libitum for 4 wk. The thoracic ducts of these rats were cannulated, and a lipid emulsion containing [3H]cholesterol and [14C]oleic acid was given through a duodenal catheter. Lymph was collected for 24 h and the isotopic tracers for cholesterol and fatty acid were measured. Rats fed the 24% sesame oil diet had significantly lower lymphatic cholesterol and fatty acid compared with the control group. Absorption of oleic acid in rats fed 24% coconut oil was significantly greater than in controls during 0-8 h but was not significantly different during 0-24 h. There were no differences among groups in the distribution of cholesterol and oleic acid either in the lymph lipoproteins or in the lipid classes. The significant reduction in lymph cholesterol and fatty acids due to sesame oil feeding may be an important factor in reducing hypercholesterolemia. PMID- 8229301 TI - Corandomization of fats improves absorption in rats. AB - Human milk fat is well absorbed despite its large concentration of long-chain saturated fatty acids (LCSFA), particularly palmitic acid. The superior absorption has been ascribed in part to the uncommonly high proportion of the palmitic acid in the sn-2 position of the triglycerides, 70% in human milk triglycerides compared with 10-15% in the fats and oils commonly used in infant formula blends. Colipase-dependent pancreatic lipase selectively hydrolyzes the fatty acids at the sn-1 and 3 positions, yielding free fatty acids and the 2 monoglyceride, which are absorbable. Free palmitic acid, but not monopalmitin, can be lost as calcium soaps in the feces. The present study demonstrated that mixtures of coconut oil and palm olein are better absorbed by rats if the proportion of LCSFA in the sn-2 position is increased by the process of chemical randomization, in which the fatty acids of the native oils are redistributed equally to all three positions in the rearranged triglycerides. The fecal excretion of total fatty acids, essentially LCSFA, from the mixtures in which the oils were randomized together (corandomized) was 30 to 60% less than from the mixtures of native oils. PMID- 8229302 TI - Mitochondrial NADPH-linked aquacobalamin reductase is distinct from the NADPH linked enzyme from microsomal membranes in rat liver. AB - Mitochondrial NADPH-linked aquacobalamin reductase was purified and characterized to clarify its enzymatic properties. The enzyme was purified about 360-fold over rat liver mitochondrial membranes in a yield of 7.5%. The purified enzyme was homogenous in SDS-PAGE. The molecular mass (M(r)) of the enzyme was calculated to be 65 kDa by SDS-PAGE and by Toyopearl HW55 gel filtration, indicating that the enzyme is a monomeric polypeptide with M(r) of 65 kDa. The enzyme was a flavoprotein containing 1 mol of FAD and FMN per mole of the enzyme. The enzyme was specific for NADPH as electron donor and had the ability to reduce cytochrome c (15.4 mumol.min-1 x mg protein-1), potassium ferricyanide (4.9 mumol.min-1 x mg protein-1) and 2,6-dichlorophenolindophenol (16.8 mumol.min-1.mg protein-1) as well as aquacobalamin (6.4 mumol.min-1 x mg protein-1). Although the enzyme immunoreacted with an antibody against NADPH-cytochrome P-450 reductase, which had the activity of the NADPH-linked aquacobalamin reductase in rat liver microsomes, the mitochondrial enzyme and the microsomal enzyme had different enzymological properties. PMID- 8229303 TI - Pyridoxine-5'-beta-D-glucoside: metabolic utilization in rats during pregnancy and availability to the fetus. AB - Pyridoxine-5'-beta-D-glucoside (PNG), which is abundant in various plant-derived food products, has been shown to be poorly utilized in male rats and adult men. To assess the metabolic utilization of PNG during pregnancy and the availability to the fetus, [3H]PNG and [14C]pyridoxine (PN) were orally administered to 15-d pregnant rats for direct assessment of the intestinal absorption, tissue distribution, and urinary excretion of the radioisotopes. The level of 3H was significantly greater in the liver, and urinary excretion of 3H tended to be lower than that of 14C. The relative distribution of 3H among the groups of B-6 vitamers within the liver was similar to that for 14C. There was greater uptake by the fetus of 3H than of 14C. There was no difference in the relative concentrations of 3H and 14C among the groups of B-6 vitamers within the fetus. Results of this study suggest that the metabolic utilization of PNG is similar to that of PN during pregnancy. Therefore, the availability of vitamin B-6 derived from maternal PNG seems to be similar to that of vitamin B-6 derived from maternal PN. PMID- 8229304 TI - High protein intake raises apparent but not true magnesium absorption in rats. AB - Earlier studies with rats have shown that greater protein intake raises apparent magnesium absorption (ingested magnesium minus fecal magnesium). We addressed the question of whether high protein intake affects true magnesium absorption. Rats were fed either a normal (175 g casein/kg) or a high (525 g casein/kg) protein diet. Extra protein was added at the expense of the glucose component of the diet; the diets were balanced for magnesium, calcium and phosphorus. The high protein diet enhanced apparent magnesium absorption. True magnesium absorption, as measured with the use of oral and intraperitoneal administration of tracer doses of 28Mg, was found to be unaffected by high protein intake. Endogenous magnesium excretion in feces was significantly depressed in rats fed the high protein diet, which was associated with elevated urinary magnesium excretion. High protein intake reduced the amount of magnesium in whole ileal digesta but raised the amount and concentration of magnesium in the liquid phase. We conclude that improvement of apparent magnesium absorption in rats fed high protein diets is due to depressed fecal excretion of endogenous magnesium. PMID- 8229305 TI - Dietary folate requirement determined for channel catfish, Ictalurus punctatus. AB - Juvenile channel catfish, Ictalurus punctatus, were fed semipurified basal diets containing 0, 0.2, 0.5, 1.0, 4.0 or 10.0 mg/kg of folic acid or 10 g/kg of succinylsulfathiazole in aquaria for 15 wk. Fish fed the sulfonamide showed higher mortality, lower weight gain, lower thrombocyte counts, higher hemocytoblast and neutrophil counts, and lower liver folate concentrations than did control fish (0 folic acid), indicating that significant intestinal bacterial synthesis of folate occurs in channel catfish. There were positive quadratic regressions of weight gain, hematocrit, erythrocyte and leukocyte numbers, and positive linear regressions of plasma and liver folate on dietary folic acid concentrations. Broken-line analysis showed that the dietary requirements for folic acid for optimum weight gain, hematocrit, and erythrocyte and leukocyte numbers were 1.01, 1.17, 1.12 and 1.15 mg/kg, respectively. Plasma and liver concentrations of folate associated with normal growth and hematopoiesis were 22.9 nmol/L and 20.0 nmol/g, respectively. Ratios of leukocytes and lymphocytes to erythrocytes were maximal in fish fed 4.0 mg folic acid/kg, indicating that immunocompetence may increase as the dietary dose exceeds that required for normal growth. Anemia in folate-deficient channel catfish was characterized by pale livers, spleens, gills and kidneys, and by poikilocytosis, anisocytosis, pyknosis, cytoplasmic clearing, increased numbers of hemocytoblasts, macrocytosis, and binucleated erythrocytes or "spectacle" cells. PMID- 8229306 TI - Dietary fiber viscosity and endogenous protein excretion at the terminal ileum of growing rats. AB - The effect of dietary fiber viscosity on the excretion of endogenous nitrogen and amino acids from the small intestinal lumen of growing rats was investigated. Rats were fed for 12 d protein-free diets containing 5% cellulose (negligible viscosity) or 5% carboxymethylcellulose of low (20 cP), medium (800 cP) or high (2000 cP) viscosity, as the sole dietary fiber source. As dietary fiber viscosity increased from 0 to 2000 cP, there was a significant (P < 0.05) linear increase in the flow of endogenous nitrogen, aspartic acid, serine, glutamic acid, proline, threonine, glycine, alanine, isoleucine, histidine, valine, methionine, leucine and lysine at the terminal ileum. The amino acid composition of the ileal digesta was unaffected by changes in fiber viscosity. It seems that either endogenous protein of a similar origin was secreted in larger amounts or that the digestion and absorption of endogenous amino acids was inhibited as dietary fiber viscosity increased. There was a significant (P < 0.05) linear increase in the concentration of sialic acids relative to chromic oxide in the small intestinal contents with increasing fiber viscosity, indicating an increase in mucoproteins. PMID- 8229307 TI - Dietary soybean protein compared with casein retards senescence in the senescence accelerated mouse. AB - The effects of replacing dietary casein with soybean protein on mean life span, mean life span of the last one-tenth of a group, grading scores of senescence and deposition of senile amyloid were investigated in senescence accelerated mice (SAM-P/1) compared with a control strain (SAM-R/1). SAM-R/1 mice fed the soybean protein-containing diet had mean life spans of 618 +/- 42 d (males) and 578 +/- 62 d (females), 58% (males) and 44% (females) longer than those of corresponding casein fed mice (P < 0.01). Similarly, in SAM-P/1 mean life-spans were 265 +/- 16 d (males) and 307 +/- 23 d (females) in the soybean diet group, 27% (males) and 30% (females) longer than in the casein diet groups (P < 0.01). The mean life span of the last one-tenth of each group fed soybean protein was significantly longer than the corresponding group fed casein. In SAM-R/1 mice, pathological studies revealed that severe secondary amyloid deposition (amyloid A protein) in the kidneys, spleen, stomach and liver was significantly suppressed, in males only, by replacing casein with soybean protein (P < 0.01). The occurrence of contracted kidneys caused by the infiltration of amyloid A protein was suppressed in SAM-R/1 mice fed the soybean protein-containing diet (P < 0.05). The deposition of senile amyloid in SAM-P/1 mice with aging was retarded by replacing casein with soybean protein (P < 0.01). These results indicate that dietary protein source is important in modulating the advance of senescence in SAM mice. PMID- 8229308 TI - Porcine somatotropin affects the dietary lysine requirement and net lysine utilization for growing pigs. AB - This study was conducted to determine the effects of exogenous porcine somatotropin (pST) on the dietary lysine requirement and efficiency of absorbed lysine utilization for pigs during the 20- to 60-kg phase of growth. Seventy-two crossbred pigs (20 +/- 0.7 kg body wt) received daily intramuscular injections of either excipient (0 dose) or pST (150 micrograms/kg body wt) and were fed diets in which protein and lysine concentrations ranged from 6.4 to 23.5 g/100 g diet and from 0.40 to 1.48 g/100 g diet, respectively. Nutrient density was altered to compensate for reduced feed intake with pST, but diets were approximately isocaloric. Rate and efficiency of gain and whole-body protein accretion rate exhibited a dose-response improvement (P < 0.01) to increases in dietary protein for both excipient and pST-treated pigs. Pigs receiving pST grew more rapidly and more efficiently than control counterparts (P < 0.01). Treatment with pST improved the rate of protein accretion (P < 0.01) at all but the lowest level of dietary protein. The net lysine utilization for lysine accretion and maintenance was 0.46 for control pigs and 0.57 for pigs receiving pST, a 24% improvement in the efficiency. Thus, treatment with pST increased the maximum rate of protein accretion as well as the partial efficiency with which dietary lysine is used for protein accretion. Consequently, only a 9% increase in dietary lysine was required to maximize protein deposition in pST-treated pigs, because the metabolic efficiency of lysine utilization was improved. PMID- 8229309 TI - Development and testing of the AIN-93 purified diets for rodents: results on growth, kidney calcification and bone mineralization in rats and mice. AB - Because of nutritional and technical problems the AIN-76A rodent diet was revised. One of the new formulations was designated AIN-93G and was suggested for use during growth, pregnancy and lactation studies. Some major differences in this new formulation compared with the AIN-76A diet are as follows: 7 g soybean oil was substituted for 5 g corn oil/100 g diet to increase the amount of linolenic acid; the amounts of vitamins E and K were increased; cornstarch was substituted for most of the sucrose; the amount of phosphorus was reduced to eliminate the problem of nephrocalcinosis in female rats; L-cystine was substituted for DL-methionine; and the manganese concentration was reduced. Various developmental modifications of the AIN-93G diet were fed to weanling rats and mice to determine effects on growth and tissue mineralization. After rats were fed the developmental version of AIN-93G for 16 wk, body weights in both male and female rats were not different from those of rats fed a cereal-based nonpurified diet. After 13 wk, male mice fed this diet weighed 13% more than those fed the nonpurified diet. Body weights of female mice were not affected. The new diet formulation prevented kidney calcification in female rats and mice during 16 wk of feeding. PMID- 8229310 TI - Taurine utilization by cats. PMID- 8229311 TI - Comment on the papers by Gonzales-Pacheco et al. (1993) and Lynn and Wallwork (1992): energy restriction and metabolic rate. PMID- 8229312 TI - AIN-93 purified diets for laboratory rodents: final report of the American Institute of Nutrition ad hoc writing committee on the reformulation of the AIN 76A rodent diet. AB - For sixteen years, the American Institute of Nutrition Rodent Diets, AIN-76 and AIN-76A, have been used extensively around the world. Because of numerous nutritional and technical problems encountered with the diet during this period, it was revised. Two new formulations were derived: AIN-93G for growth, pregnancy and lactation, and AIN-93M for adult maintenance. Some major differences in the new formulation of AIN-93G compared with AIN-76A are as follows: 7 g soybean oil/100 g diet was substituted for 5 g corn oil/100 g diet to increase the amount of linolenic acid; cornstarch was substituted for sucrose; the amount of phosphorus was reduced to help eliminate the problem of kidney calcification in female rats; L-cystine was substituted for DL-methionine as the amino acid supplement for casein, known to be deficient in the sulfur amino acids; manganese concentration was lowered to one-fifth the amount in the old diet; the amounts of vitamin E, vitamin K and vitamin B-12 were increased; and molybdenum, silicon, fluoride, nickel, boron, lithium and vanadium were added to the mineral mix. For the AIN-93M maintenance diet, the amount of fat was lowered to 40 g/kg diet from 70 g/kg diet, and the amount of casein to 140 g/kg from 200 g/kg in the AIN-93G diet. Because of a better balance of essential nutrients, the AIN-93 diets may prove to be a better choice than AIN-76A for long-term as well as short-term studies with laboratory rodents. PMID- 8229313 TI - Determination of alpha-tocopherol stereoisomers in biological specimens using chiral phase high-performance liquid chromatography. AB - The alpha-tocopherol stereoisomers in biological specimens were investigated using high-performance liquid chromatography. All-rac-alpha-Toc acetate was separated into four peaks (peak area ratio: 4:2:1:1) by Chiralpak OP(+)HPLC. 2R isomers constituted the first peak and 2S-isomers were separated into three peaks (peak area ratio: 2:1:1). 2-Ambo-alpha-Toc acetate was completely separated into RRR- and SRR-alpha-Toc acetate by this method. The present HPLC method was used for the separation of all-rac-alpha-Toc in blood and tissues of rat. The analytical recoveries of RRR- and SRR-alpha-Toc acetate added to blood and tissues were 90.5-98.2% for RRR-alpha-Toc and 93.7-100.5% for SRR-alpha-Toc. The distribution of alpha-Toc stereoisomers in the blood and tissues from rats administered all-rac-alpha-Toc acetate was investigated by HPLC. The concentrations of the 2R-isomers in the blood and tissues were markedly higher than the concentrations of the 2S-isomers, and the levels of alpha-Toc stereoisomers showed marked differences between blood and tissues. PMID- 8229314 TI - New retinoids inhibiting the growth of human acute promyelocytic leukemia cell line HL-60 without inducing differentiation. AB - A series of acid homologs with longer side chain length was synthesized and their biological activities to inhibit growth and to induce differentiation of human acute promyelocytic leukemia cell line HL-60 were analyzed. It was found that ethyl alpha-retinylidene propionate (E) shows interesting activity. This compound inhibited growth of HL-60 cells with almost the same potency of retinoic acid. However, unlike retinoic acid, the potency of this compound to induce differentiation of HL-60 cells into neutrophiles was almost negligible. Ethyl alpha-retinylidene propionate (E) is an unprecedented retinoid which inhibits growth of HL-60 cells without inducing cell differentiation. The Z isomer [ethyl alpha-retinylidene propionate (Z)] also showed similar unique effects on HL-60 cells. Interestingly, both of the E and Z isomers of alpha-retinylidene propionic acid showed only very weak activities to inhibit growth and to induce differentiation of HL-60 cells. It was suggested that ethyl alpha-retinylidene propionate acts on HL-60 cells in the intact form without suffering hydrolysis, and also that the ester group plays important role for the unique activity. Biological activities of retinylidene acetic acid (E and Z) which lacks methyl group at alpha-position of alpha-retinylidene propionic acid, on HL-60 cells were weaker than those of the isomers of alpha-retinylidene propionic acid. Esterification of the isomers of retinylidene acetic acid resulted in further decrease of biological activities. Biological activities of ethyl retinoate on HL 60 cells were also very weak. It was indicated that methyl group present at alpha position of ethyl alpha-retinylidene propionate plays an important role for the unique biological activity. It was further indicated that the effect of esterification of carboxyl group of retinoids on the biological activity is not constant, but varies depending on the structure. Pretreatment of HL-60 cells with ethyl alpha-retinylidene propionate (E) apparently did not influence the process of induction of differentiation of HL-60 cells into neutrophiles by retinoic acid. It was further shown that ethyl alpha-retinylidene propionate (E) prolongs the time necessary for induction of differentiation of HL-60 cells by retinoic acid, but does not influence the level of final rate of differentiation. PMID- 8229315 TI - Drinking habits influence the relationship between apolipoprotein AII and body mass index. AB - The relationships of cigarette smoking, exercise, and body mass index (BMI) to the serum levels of high-density lipoprotein (HDL) cholesterol, apolipoprotein AI (Apo AI), apolipoprotein AII (Apo AII), and % of HDL cholesterol to total cholesterol (%HDL-C) were assessed in 495 employees of a telephone company in Japan. They were subdivided into four groups by frequency of alcohol intake: nondrinkers (n = 28), those who drank once per week (n = 78), those who drank 2 to 5 times per week (n = 245), and those who drank 6 to 7 times per week (n = 144). Although univariate analysis before grouping showed a positive correlation between Apo AII and BMI, multiple analysis examined in each group showed the positive relationship only in the nondrinking group. The relationship between BMI and Apo AII would thus appear to depend on drinking habits. PMID- 8229316 TI - Growth and essential fatty acid levels of plasma and liver of suckling rats from mothers fed on fat-free or various fat diets. AB - The milk fatty acid compositions of mothers fed on a fat-free or various fat diets, and the effects on growth and fatty acid compositions of their pups were studied. Even the milk of essential fatty acid-deficient mothers fed on a fat free or hydrogenated fat diet contained about 3 and 1.7%, n-6 and n-3 fatty acids, respectively. In the plasma of the suckling pups, however, the proportions of n-6 and n-3 fatty acids rapidly increased to about 20 and 3-5%, respectively, at 1 week after birth. In particular, the PUFAs markedly increased in the liver PC and PE, and the high levels were maintained until weaning. Although the PUFA compositions of suckling pups were influenced with those of maternal diet, small amounts of n-6 and n-3 fatty acids were usually maintained in the plasma and liver. After weaning to the same diets (without PUFAs) as the mothers, however, the n-3 and n-6 fatty acids rapidly decreased and endogenous n-9 eicosatrienoic acid appeared. On the other hand, the growth during suckling was not significantly different among the litters of mothers fed on diets with or without n-3 or n-6 fatty acids. After the weaning, however, the growth was improved in the following order: corn oil, perilla oil > fish oil > fat-free, hydrogenated fat diet group. n-3 fatty acids appeared to be used partially as substitutes for n-6. However, the essentiality was not clear, as the n-3 fatty acids always coexisted with the n-6. Thus, it appeared that small amounts of n-3 and n-6 fatty acids in milk were supplied to the suckling animals regardless of maternal diet and supported growth. PMID- 8229317 TI - Effects of endogenous and exogenous n-3 and n-6 fatty acids on microsomal synthesis of docosahexaenoic acid in vitro. AB - Effects of endogenous and exogenous polyunsaturated fatty acids (PUFA) on microsomal synthesis of docosahexaenoic acid (DHA) from eicosapentaenoic acid (EPA) in vitro were studied using rat livers. Liver microsomes prepared from three groups of rats which had been fed n-3/n-6 ratios of 0.01, 0.39, and 2.70, respectively, for 1 month, were incubated with [1-14C]EPA at 37 degrees C for 30 min. There were no significant differences between the formations of docosapentaenoic acid (DPA) or DHA in the groups. Liver microsomes from rats fed a diet with an n-3/n-6 ratio of 2.70 for 3 weeks were then incubated with [1 14C]EPA in the presence of unlabeled DHA or arachidonic acid (AA). The additional DHA did not have any effect. However, a significant inhibition of DHA synthesis was observed by addition of AA (EPA/AA = 5:2). These results suggest that the microsomal biosynthesis of DHA in rat liver is not susceptible to feedback inhibition. PMID- 8229318 TI - Dietary fiber intake of Japanese younger generations and the recommended daily allowance. AB - A method was introduced for the estimation of total dietary fiber (TDF) intake of a population using a menu-oriented questionnaire and a menu-based calculation table. TDF intake correlated well with age in a population investigated, and in younger generations TDF consumption was very low (less than 11.5 g/day in teenagers). The similar results were obtained from the calculation using data of National Nutrition Survey (10.7 g/day). The foodstuffs they consumed were more processed and refined. This fact suggested that in younger generations a future resumption of their present eating habits might produce a serious lack of TDF intake in later years. To clarify the optimal level of TDF intake for the upper limit of recommended daily allowance (RDA) of an average Japanese, the following were measured and calculated. (I) Estimation of recent TDF intake (1990) and of 30 and 50 years ago (1955, 1935), based on TDF data of foodstuffs by the enzymatic-gravimetric method. (II) Measurement of the TDF of model duplicate meals and model composite diets for the average Japanese in 1985 using the same assay method. (III) Conversion of a recommendation of 20-35 g/day for American into RDA for Japanese considering energy consumption and lower fat intake. (IV) Re-estimation of the literature data on the adverse effects of DF on the human mineral balance considering the insufficient calcium intake of Japanese. The results indicated an RDA of 10-12 g TDF/1,000 kcal fit better for an average Japanese. PMID- 8229319 TI - Effects of transgalactosylated disaccharides on the human intestinal microflora and their metabolism. AB - The effects of transgalactosylated disaccharide (TD) intake on human fecal microflora and their metabolism were investigated in 12 Japanese males. TD is a mixture of sugars, galactosyl galactose, and galactosyl glucose, synthesized from lactose through the transgalactosylation reaction of Streptococcus thermophilus beta-galactosidase. Volunteers took 15 g of the test sugar daily for 6 days. The TD ingestion increased the number of bifidobacteria and lactobacilli, but decreased the number of Bacteroidaceae and Candida spp. in the feces. The ratio of bifidobacteria to total bacteria increased from 0.28 to 0.51. TD decreased the fecal concentrations of propionic acid, isobutyric acid, isovaleric acid, and valeric acid. This sugar also lowered the fecal pH, and the concentrations of fecal ammonia, p-cresol, and indole. Moreover, a positive correlation was found between the concentration of ammonia, and that of branched-chain fatty acids (isobutyric acid and isovaleric acid), p-cresol, and indole. All of these compounds are produced from amino acids through deamination by the intestinal bacteria. The depression of amino acid fermentation by intestinal bacteria may be involved in the reduction of fecal ammonia. These results suggest that a part of the transgalactosylated disaccharides passes into the colon, inducing changes in the colonic microflora composition, hastening carbohydrate fermentation, and depressing amino acid fermentation in the human gut. PMID- 8229320 TI - What is the Committee's response to management personnel who wish to ease first aid and CPR training in a small plant (100 employees) because of the potential cost of complying with OSHA's blood-borne pathogen standard? The facility primarily employs engineers and technicians involved in research development. PMID- 8229321 TI - Our plant's employees are potentially exposed to elemental mercury. What are signs and symptoms of potential exposure and what is the significance of biological monitoring? PMID- 8229322 TI - "Sick building syndrome": an inappropriate term. PMID- 8229323 TI - Occupational physicians and environmental medicine. PMID- 8229324 TI - Electrocutions involving hand-held power tools. PMID- 8229325 TI - Concurrent validity of the ERGOS work simulator versus conventional functional capacity evaluation techniques in a workers' compensation population. AB - A prospective blinded cohort study was performed in an interdisciplinary vocational evaluation program to investigate the concurrent validity of the ERGOS work simulator in comparison to current methods of evaluation. Seventy men and eight women, aged 22 to 64 years, who attended for a 2-week physical capacity assessment participated in the study. Physical activity factors as defined by the Canadian Classification and Dictionary of Occupations and the American Dictionary of Occupational Titles were assessed for all subjects under three evaluation conditions: the ERGOS work simulator, an exercise-oriented physical evaluation by a rehabilitation therapist, and performance of project-format industrial tasks. In addition, 17 men and 7 women were assessed with VALPAR standardized work sample tests. The statistical significance of the relationships between results obtained by the various evaluation methods was examined. There was a strong correlation between the ERGOS dynamometry and the clinical assessment of strength for all standard movements tested (P < .001). The Methods Time Measurement rating by the ERGOS for dexterity variables, according to industrial engineering standards, tended to rate subjects as more restricted than did the clinical evaluators. There was a significant relationship (P < .001) between the "overall physical activity rating" from ERGOS dynamometry, clinical evaluation, and performance in an industrial workshop setting. There was also a significant relationship (P < .001) between the "overall physical activity rating" for endurance of a full workday produced by the 4-hour ERGOS evaluation and by the 2 week functional capacity evaluation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229326 TI - Occupational medicine training program surveys. AB - Although the quantity of Occupational Medicine (OM) residents in training programs has risen in the past 15 years, there remains a significant shortage of OM physicians in the United States. A survey of OM residency program directors and residents and fellows (trainees) was accomplished to answer questions relevant to recruiting and supplying OM trained physicians. Twenty-six of 29 program directors (89.7%) replied. One hundred forty-three of 174 (82.2%) trainees responded. Fifty percent of responding program directors were satisfied with the current quantity of program applicants. Medical students were provided a median of 3 hours (range, 0 to 20 hours; mean, 5.35 hours) of formal OM teaching by the responding programs. Almost half of trainees, 68 (48.6%), did not receive formal OM instruction in medical school. An average of 5.4 +/- 13.3 hours of OM was taught to OM trainees in medical school. Disproportionate numbers of trainees were taught OM as fourth-year medical students and in other residencies. Reasons for pursuing OM training were diverse and often related to postgraduate clinical experience with OM. Only a total of 84 primary care residents (0.32%) rotate through the OM residency programs in an average year. We conclude that an insufficient quantity of qualified applicants, combined with limited exposure to OM in medical schools and low levels of contact with residents in primary care programs, will continue to hinder efforts to reduce the shortage of OM residents and physicians. Further training specifically targeting the fourth year of medical school and the primary care residencies may have the most impact on recruitment. PMID- 8229327 TI - Hematologic effects of benzene. Job-specific trends during the first year of employment among a cohort of benzene-exposed rubber workers. AB - Hematologic surveillance data from 1940 to 1975 were analyzed for a benzene exposed cohort of 459 rubber workers. The present analyses are restricted to 161 workers with "preemployment" counts done before exposure and rely on their subsequent counts from the first 12 months of employment. While blood cell counts declined approximately 1000 cells/mm3 over the first 4 months of exposure. Using repeated-measures analysis of variance, workers exposed above the median benzene exposure at the plant had significantly lower average white and red blood cell counts at each month during the first year of work when compared with workers exposed below the median. These decreased counts suggest that clinically detectable bone marrow depression accompanied the onset of work in this plant during the 1940s and support exposure assessments that favor higher benzene levels in the 1940s when compared with subsequent decades. The general utility of repeated-measures analytic techniques for medical surveillance data is also demonstrated by this analysis. PMID- 8229328 TI - Actions and reactions. Effects of employee theft on worker health. AB - The health and safety problems that arise from employee theft are considerable and should be recognized. Effective prevention of employee theft should focus on employee selection, education, and the factors associated with theft such as drug and alcohol abuse. When physical barriers are used at work sites to prevent theft, work site egress should be safe. Interaction between management and occupational health professionals can focus attention on the relationship between work site theft and employee health. PMID- 8229329 TI - Consistency of sincere and feigned grip exertions with repeated testing. AB - Our purpose was to test the hypothesis that feigned exertions would show greater variability in both force and electromyograms than would sincere exertions over repeated testing sessions. Eleven normal subjects made sincere and feigned exertions while performing the five-handle position Jamar grip test. The subjects were tested 6 times in a 3- to 5-week period. Contrary to the hypothesis, feigned exertions did not show greater variability than sincere exertions. Both sincere and feigned exertions were highly consistent over the six sessions. However, the sincere and feigned exertions did differ in the patterns of force and electromyogram on the five-handle position test. These results and those of previous studies suggest that clinicians should use more than one type of test when testing suspected malingerers. PMID- 8229330 TI - Effectiveness of self-help smoking cessation guides for firefighters. AB - Most smokers would prefer to quit smoking on their own. However, among specific blue-collar occupations it is not known whether self-help techniques will be effective or even used to help smokers quit smoking. In this study we evaluated the effectiveness of a self-help smoking cessation guide designed specifically for firefighters (FF) and compared it with the American Lung Association (ALA) guide designed for the general public. We hypothesized that a shorter guide (FF), written in the jargon of the fire service, and formatted as a workbook would be more effective. One hundred five firefighters randomly received either the ALA or FF guide after having an annual physical examination. Twelve percent of the ALA group and 11% of the FF group quit smoking after the physical examination but before receiving the guides. We found no relationship between the specific guide that the firefighters received and subsequent smoking status. Few firefighters reported filling out pages of the ALA guide (11.9%) or the FF guide (17.8%). For smokers who have not reached the active stage of quitting smoking, guides may not be a factor in quitting. Screening smokers to determine their "stage of change" at the physical examination may increase the effectiveness of self-help guides. PMID- 8229331 TI - A work-site weight management program to reinforce behavior. AB - This pilot weight management project addresses the efficacy of reinforcing dietary behavior change versus weight loss. This 6-month program served professional and support staff participating in the work-site wellness program at a midwestern university. Behavior-contingent program data were compared with data from the previous model where contracts were made for weight loss. In the behavior-contingent program, dropout rate and satisfaction with the program compared favorably with the old model weight loss-contingent program. Contract adherence was 93% compared with 74% in the weight loss-contingent program. Actual pounds of weight lost were lower in the behavior-contingent program, however, long-term weight management must still be studied with this population. PMID- 8229332 TI - Supermarket checker motions and cumulative trauma risk. AB - The relationship between specific motions and symptoms consistent with upper extremity cumulative trauma disorders (UECTDs) was investigated in 50 supermarket checkers. Each completed a questionnaire concerning UECTD symptoms; four composite symptom indices were derived: any arm symptoms, hand-wrist-lower arm (S DIST), upper arm, and symptoms specifically associated with carpal tunnel syndrome (S-SPC). Each participant was videotaped on at least two occasions while performing checking work. Analysis of these tapes assigned each subject a composite motion index for each of the following motions: wrist flexion, wrist extension, body (lumbar) flexion, pronation, grip type, and tendency to lift objects. "Positive" symptom status was defined by a score in the upper quartile for the symptom index. Relationships between an individual's motion indices and symptom indices were analyzed by determining the percent of subjects "positive" for symptoms in each quartile of motion index, by rank correlation, and by regression of symptom scores on principal components of motions. Trends toward relationship of wrist flexion and extension, lumbar flexion, and pronation with S DIST and S-SPC were noted. Principal components regression confirmed that extension and flexion were related to these two symptom outcomes. This study suggests that postural loading can be determined on an individual basis in a meaningful fashion and that interventions that decrease such loading may be beneficial. It supports the role of certain repetitive motions as causes of UECTD symptoms. PMID- 8229333 TI - Measuring the climate for health at organizations. Development of the worksite health climate scales. AB - Worksite health promotion research has overemphasized the impact of individual behaviors on employee well-being and neglected the important influence of the work environment. In the present research effort, measures of the health climate at the worksite were developed, administered to employees at a newspaper company, and then tested for their psychometric properties. After revising the original scales, several health outcome variables and an improved version of the measure were administered to employees at seven small worksites. The results indicated that the health climate differed significantly across worksites and that health climate perceptions were significantly related to measures of physical symptoms; exercise, nutrition, and smoking habits; job stress; and job satisfaction. PMID- 8229334 TI - Disability: a new psychosocial perspective. AB - A previously disabled family member is presented as a psychosocial determinant in the subsequent disability of the patient. Eighty-three disabled patients presented for an independent medical examination of the upper extremity. All were seeking a workers' compensation settlement or continued workers' compensation benefits. Patients were questioned regarding the current disability of any family member(s). Forty-four of the 83 had at least one family member who was disabled and 36 had experienced a traumatic incident or injury resulting in workers' compensation. Forty-seven had no objective impairment. These findings suggest a significant correlation between the "disabled" support system and subsequent disability of the patient (P < .001). PMID- 8229335 TI - What is your recommendation concerning the vaccination of an employee who will be traveling for 3 months in a country that is considered a rabies-endemic area? PMID- 8229336 TI - What is the appropriate treatment regimen for a lead-exposed employee with a blood lead level of 60 micrograms/dl? PMID- 8229337 TI - Primary cutaneous carcinoma of the scrotum. PMID- 8229338 TI - Where do occupational medicine residencies fit in the institution? PMID- 8229339 TI - Definitions, sources, magnitude, effect modifiers, and strategies of reduction of the healthy worker effect. PMID- 8229340 TI - Epidemiology update. PMID- 8229341 TI - Coal, lead, asbestos, and HIV. The politics of regulating risk. PMID- 8229342 TI - Cancer risk assessment and management. An ethical perspective. AB - We explored ethical issues in cancer risk assessment and management. Although literature about risk assessment and management is increasing, few attempts have been made to address directly either their ethical aspects or implications. We have compared ethical considerations raised at the community level with those classically considered in the physician-patient setting. Established principles of autonomy, beneficence, nonmaleficence, and distributive justice are shown not to be easily applicable at the societal level. Available information about the safety of exposures often is too scanty to allow decisional autonomy; beneficence is considered from the societal viewpoint rather than from that of the individual; and equity is frequently violated. Ethics require careful consideration when defining what is acceptable to society; the simple mathematical formulations used for risk-benefit evaluations cannot be applied directly to individuals. Further development and integration of these ideas are necessary. PMID- 8229343 TI - Asthma related to occupational and ambient air pollutants in nonsmokers. AB - We attempted to determine the association between occupational and air pollutant exposure with the development of adult asthma through the analysis of a standardized respiratory questionnaire administered to a cohort of 3914 nonsmoking adults in 1977 and again in 1987. Ambient air pollution concentrations were estimated over a 20-year period using monthly interpolations from fixed-site monitoring stations applied to zip code locations by month of residence and work site. Second-hand smoke exposure was significantly associated with the development of asthma (related risk [RR] = 1.45, confidence interval [CI] = 1.21 to 1.75). Airways obstructive disease before age 16 was related to a marked increased risk (RR = 4.24, CI = 4.03 to 4.45). An increased risk of asthma was significantly associated with increased ambient concentrations of ozone exposure in men (RR = 3.12, CI = 1.61 to 5.85). PMID- 8229344 TI - Fatal occupational injuries in the New Jersey construction industry, 1983 to 1989. AB - Work in the construction industry involves about a threefold increased risk of fatal injury compared with all industries combined. The purpose of this study was to identify potential risk factors for fatal injury in the construction industry in New Jersey. Multiple data sources including death certificates, medical examiner reports, Occupational Safety and Health fatality files, and Workers' Compensation reports were used to identify 200 construction-related fatalities in New Jersey during the years 1983 to 1989. All deaths were in men. The death rate was 14.5 per 100,000 employed person-years over the study period. Death rates tended to diminish with increasing age after 34 until age 65 when the death rate was the highest (27.7). Death rates were higher for Hispanics (34.8) and African Americans (24) than whites (10.6). Ironworkers and roofers had highest rates (109.0 and 56.2, respectively) among specific occupational groups within the construction industry. The leading cause of death was falls (46%). These data suggest that intervention efforts directed toward workers at heights is needed. Further research is warranted to elucidate the factors contributing to the elevated fatality rate of workers over age 65, and to Hispanic and African American workers. PMID- 8229345 TI - Mobile Work Site Health Promotion Programs can reduce selected employee health risks. AB - This study examined the effect of participation in a mobile work site health promotion program (MWHPP) on selected health risks. Measures of blood pressure, cholesterol total and total/high-density lipoprotein ratio, percent body fat, and submaximal fitness were taken at baseline, 6-, 12-, and 18-month intervals from 113 employed adults. Significant reductions (P < or = .01) were observed in all variables measured. Further analysis of the number of employees who went from high risk to low risk demonstrated a significantly reduced risk of hypercholesterolemia, cholesterol total/high-density lipoprotein ration, and obesity (average significant chi 2 = 9.3, P < or = .05). It was concluded that participation in MWHPPs can significantly reduce several major cardiovascular risk factors. MWHPPs are relatively inexpensive compared with in-house health promotion programs and are one of the most cost-effective methods of reducing employee health risk. PMID- 8229346 TI - Relationship between occupation and episodes of headache that match cervical origin pain patterns. AB - The relationship between headache that matches cervical origin pain patterns, occupation, gender, age, and hours of work is examined in a randomly sampled, never-injured population. Although gender was established as a significant factor in the association between occupation and headache, age and hours of work were shown to have no effect. Women working in managerial and professional occupations had a significantly higher risk of cervical origin headache when compared with women working in either clerical or blue collar occupations. A similar association for men was not noted. PMID- 8229347 TI - Hand-arm vibration syndrome: a guide to medical impairment assessment. AB - Hand-arm vibration syndrome is a complex condition with vascular, sensorineural, and musculoskeletal components. Workers who handle vibratory tools suffer from it, and the severity of this syndrome is now graded internationally using the Stockholm classification. For compensation purposes the severity must be translated into impairment and this paper proposes how this may be done using the AMA Guides to the Evaluation of Permanent Impairment. The case history of a typical claimant is presented to demonstrate how an impairment rating might be derived. PMID- 8229348 TI - Pregnancy outcome among women exposed to pesticides through work or residence in an agricultural area. AB - The study population consisted of women enrolled in a southern California community clinic perinatal program. All women entering the program between January 1987 and December 1989 were asked to participate, and a cohort of 535 women was included in the study. Primarily Hispanic, of low income and educational level, many had recently immigrated from Mexico. They were potentially exposed to pesticides and other agricultural chemicals occupationally and/or environmentally because agricultural production in San Diego County is among the highest in the state. Study methods consisted of biologic assay of maternal blood samples for determination of cholinesterase activity and concurrent patient interviews to determine exposure history by self report. These assessments were conducted on each participant approximately once each trimester. Self-report and blood assay data were analyzed using chi 2, the Mantel-Haenszel extension of chi 2, and risk ratios to determine the association between pesticide exposure and spontaneous abortion, preterm birth, low birth weight and toxemia. No difference between exposed and unexposed women was noted for risk of preterm birth or toxemia. Subjects who experienced spontaneous abortion were all unexposed, and the rate of spontaneous abortion was 2.1%, less than generally expected. A greater incidence of low birth weight among unexposed women indicates that exposure may have had a "protective" effect. PMID- 8229349 TI - Mortality among employees of a perfluorooctanoic acid production plant. AB - Perfluorooctanoic acid (PFOA) has been found at low levels (10 to 100 parts per billion) in sera of the general population and at higher levels in occupationally exposed workers. Although PFOA has been reported to be a promoter of rodent hepatocarcinogenesis and to alter reproductive hormones in humans and rodents, there is little information on human health effects associated with PFOA exposure. The present study examined the relationship between PFOA and mortality using a retrospective cohort mortality design. The cohort consisted of 2788 male and 749 female workers employed between 1947 and 1983 at a plant that produced PFOA. The all-causes standardized mortality ratio was .75 (95% confidence interval [CI], .56 to .99) for women and .77 (95% CI, .69 to .86) for men. Among men the cardiovascular standardized mortality rate was .68 (95% CI, .58 to .80) and the all-gastrointestinal diseases was .57 (95% CI, .29 to .99). There was no significantly increased cause-specific standardized mortality ratio for either men or women. Ten years of employment in exposed jobs was associated with a 3.3 fold increase (95% CI, 1.02 to 10.6) in prostate cancer mortality compared to no employment in PFOA production. There were only six prostate cancer deaths overall and four among the exposed workers; thus, the results must be interpreted cautiously. If prostate cancer mortality is related to PFOA, PFOA may increase prostate cancer mortality by altering reproductive hormones in male workers. PMID- 8229350 TI - Aberration-free measurements of the visibility of isoluminant gratings. AB - We developed a new apparatus and psychophysical technique to extend isoluminant contrast-sensitivity measurements to high spatial frequencies. The apparatus consists of two identical laser interferometers that are designed to produce phase-locked two-color interference fringes on the retina without the influence of diffraction and most aberrations in the eye. However, even with interferometry, transverse chromatic aberration of the eye can produce a wavelength-dependent phase shift in the interference fringes, which can be exaggerated by head movements. To reduce the effect of head movements, isoluminant red and green interference fringes of equal spatial frequency and orientation were drifted slowly in opposite directions to guarantee a purely isochromatic (in phase) and a purely isoluminant (out of phase) stimulus during each cycle of stimulus presentation. With this technique we found that observers could resolve red and green stripes at spatial frequencies higher than 20 cycles per degree (c/deg) (20-27 c/deg), substantially higher than has previously been reported. This places a lower bound on the sampling density of neurons that mediate color vision. At all spatial frequencies, even those above the isoluminant resolution limit, a relative phase of the red and the green components could be found that obliterated the appearance of luminance modulation at the fringe frequency. Above the resolution limit, red-green-isoluminant interference fringes are seen as spatial noise, which may be chromatic aliasing caused by spatial sampling at some stage in the chromatic pathway. PMID- 8229351 TI - Efficiency in detection of isoluminant and isochromatic interference fringes. AB - We examined the limitations imposed by neural factors on spatial contrast sensitivity for both isochromatic and isoluminant gratings. We used two strategies to isolate these neural factors. First, we eliminated the effect of blurring by the dioptrics of the eye by using interference fringes. Second, we corrected our data for additional sensitivity losses up to and including the site of photon absorption by applying an ideal-observer analysis described by Geisler [J. Opt. Soc. Am. A 1, 775 (1984)]. Our measurements indicate that the neural visual system modifies the shape of the contrast-sensitivity functions for both isochromatic and isoluminant stimuli at high spatial frequencies. If we assume that the high-spatial-frequency performance of the neural visual system is determined by a low-pass spatial filter followed by additive noise, then the visual system has a spatial bandwidth 1.8 times lower for isoluminant red-green than for isochromatic stimuli. On the other hand, we find no difference in bandwidth or sensitivity of the neural visual system for isoluminant red-green and S-cone-isolated stimuli. PMID- 8229352 TI - How much does illuminant color affect unattributed colors? AB - Does the light coming from a surface (as opposed to the surface color) appear the same after adaptation to a new illumination as it did before the illumination changed? Many answers have been proposed over the past century, but all have been unsatisfactory. The main measurement problem is to provide a comparison stimulus that is unaffected by the adaptation being tested. My observers used a mental standard. Under 4000-, 6500-, and 10,000-K adaptations (the extremes and the average of daylight) they produced on a CRT unique hues with a constant saturation that was memorized during training. The main evaluation problem is how to determine the theoretical chromaticity shifts that represent illumination invariance for comparison with the data. Like most previous investigators, I used light sources rather than actual surfaces and illuminants. Using a new technique, I determined theoretical surfaces that would have unique hues under the test illuminants. Using Cohen's basis vectors, I derived theoretical reflectances that under 6500 K would produce the chromaticities that the observer chose as unique hues. The chromaticities of those same reflectances under 4000 and 10,000 K are theoretical points representing illuminants-invariant appearance of the light coming from the surfaces. Even for this small range of illuminants the adaptive shifts were too small for invariance, i.e., the appearance of the light was different even after full adaptation. This result sharpens the question of the basis for humans' concept of color as a stable property of surfaces. PMID- 8229353 TI - Color constancy. I. Basic theory of two-stage linear recovery of spectral descriptions for lights and surfaces. AB - Changing a scene's illuminant causes the chromatic properties of reflected lights to change. This change in the lights from surfaces provides spectral information about surface reflectances and illuminants. We examine conditions under which these properties may be recovered by using bilinear models. Necessary conditions that follow from comparing the number of equations and the number of unknowns in the recovery procedure are not sufficient for unique recovery. Necessary and sufficient conditions follow from demanding a one-to-one relationship between quantum catch data and sets of lit surfaces. We present an algorithm for determining whether spectral descriptions of lights and surfaces can be recovered uniquely from reflected lights. PMID- 8229354 TI - Color constancy. II. Results for two-stage linear recovery of spectral descriptions for lights and surfaces. AB - Our analysis of color constancy in a companion paper [J. Opt. Soc. Am A 10, 2148 (1993)] provided an algorithm that lets one test how well linear color constancy schemes work. Here we present the results of applying the algorithm to a large parametric class of color constancy problems involving bilinear models that relate photoreceptoral spectral sensitivities, surface reflectance functions, and illuminant spectral power distributions. These results, supported by simulation and further analysis, provide a detailed classification of two-stage linear methods for recovering the spectral properties of reflectances and illuminants from reflected lights. PMID- 8229355 TI - On wheezing insurance. PMID- 8229356 TI - Classification of autogenous skull grafts in cranial reconstruction. AB - We have presented a classification of autogenous skull grafts in cranial reconstruction: transpositional, sliding, full thickness split, shave, bone dust, and vascular pedicle. The advantages, uses and disadvantages are discussed. It is shown that each type has characteristics that make it the treatment of choice for cranial defects in specific areas. However, a combination of these techniques also can be used in dealing with some cranial defects. PMID- 8229357 TI - Diseases associated with pet and wild birds. PMID- 8229358 TI - Symposium on antimicrobial therapy. X. Chloramphenicol, clindamycin, and metronidazole. PMID- 8229359 TI - Laparoscopic gastropexy as treatment for incarcerated and obstructing sliding hiatal hernia. PMID- 8229360 TI - 'Tis the season for venomous bites, stings. PMID- 8229361 TI - Patient details his life-and-death struggle with health care regs. PMID- 8229362 TI - Awesome duty. PMID- 8229363 TI - Symposium on antimicrobial therapy. XI. Antituberculosis agents. PMID- 8229364 TI - Laparoscopic repair of perforated duodenal ulcer by Graham omentopexy. PMID- 8229365 TI - But who will guard the guardians themselves? PMID- 8229366 TI - Remarks of the First Lady to the American Medical Association. PMID- 8229367 TI - Leaders in medicine. John A. McIntyre, MD. PMID- 8229368 TI - Rapid AIDS test still controversial. PMID- 8229369 TI - Who does this patient belong to? PMID- 8229370 TI - Clinton's Tulsa disclosures. PMID- 8229371 TI - Methicillin-resistant Staphylococcus aureus: long-term routes of acquisition in hospital patients. AB - Five hundred fifty three patients and two personnel (1986-1991) were categorized according to acquisition route of methicillin-resistant Staphylococcus aureus (MRSA) and initial culture date. Of the 555, there were 218 with nosocomial infections, 100 with nosocomial colonizations, 92 with community-acquired infections, and 145 with community-acquired colonizations. Nosocomial infections occurred significantly more often than nosocomial colonizations, community acquired infections, or colonizations (p < 0.001). Results of this study show that over a long period, our patients acquired MRSA by different routes, none of which were related to seasons of the year. Although seasonal outbreaks of epidemic MRSA are important, an awareness of the occurrence of endemic MRSA is also important because it serves as a reminder that MRSA reservoirs are continuously present in some institutions. PMID- 8229372 TI - Wilson's disease: a diagnosis made in two individuals greater than 40 years of age. AB - Wilson's disease is an autosomal recessive disease of copper metabolism which is widely recognized as a disease occurring clinically in children, adolescents, and young adults. Unrecognized and therefore untreated Wilson's disease in patients over age 40 is thought to occur either rarely or not at all. Two cases of Wilson's disease presenting at an age greater than 40 years are presented. The first is a 42-year-old Israeli women who presented with fulminant hepatic failure. The serologic and biochemical investigations obtained at the time of her fulminant hepatic failure included copper studies which suggested the diagnosis of Wilson's disease, which was confirmed by an examination of the native liver following successful orthotopic liver transplantation. The second case is that of a 56-year-old white male who presented to the hospital with a three-year history of neurological dysfunction, pancytopenia, and mild splenomegaly. A battery of serologic and biochemical investigations suggested a diagnosis of Wilson's disease. The diagnosis was confirmed by quantitative hepatic copper estimation and the demonstration of Wilson's disease in three of his siblings, all of whom were diagnosed after the proband case had been identified. This man and his siblings have been treated with d-penicillamine, with remarkable improvement in their neurologic and hepatic function. The proband is currently well 11 years after his diagnosis was established. These two cases demonstrate that a diagnosis of Wilson's disease should be considered as part of the differential diagnosis of individuals in the fourth and fifth decades of life who present with unexplained liver disease. PMID- 8229373 TI - Domestic violence--battered women syndrome. AB - Domestic violence is extremely widespread and often goes unrecognized by physicians. The problem is discussed and ideas for dealing with it are outlined. PMID- 8229374 TI - National Network of Libraries of Medicine and health science libraries in Oklahoma. AB - Rural physicians have access to a considerable variety of medical library resources, despite their outlying practice locations. This article outlines some of the sources available and how they may be reached. PMID- 8229375 TI - Patient-oriented functional outcome after unilateral lower extremity fracture. AB - To determine patient-perceived functional outcome after lower extremity fracture (LEF), a prospective, follow-up study of patients managed at three level I trauma centers was conducted. Patients with unilateral LEF involving the acetabulum and distally were eligible for the study. A total of 444 patients were enrolled. Of these, 363 (82%) were interviewed at 6 months postdischarge. Study patients were predominantly young (mean age 34 years), white (72%) men (71%) who had been working preinjury (78%). Their injuries resulted primarily from motor vehicle crashes (73%); 30% had more than one fracture to the same extremity. Functional status was measured using the Sickness Impact Profile (SIP), a well-validated, general health status instrument. Mean 6-month SIP scores were significantly worse (higher) than those based on preinjury activities (9.8 vs. 2.5) (p < 0.01). Overall disability levels were moderate compared with other health conditions. Analysis of the 12 subscores comprising the SIP indicated particularly high scores in ambulation (16.2 postdischarge vs. 1.1 preinjury), sleep/rest (13.1 vs. 5.1), household management (14.5 vs. 2.6), recreation (17.6 vs. 4.2), emotional well-being (9.9 vs. 2.1), and most significantly work (33.2 vs. 8.8). Of those working preinjury, only 49% had returned by 6 months. SIP scores were highest for persons with three or more fractures to the same extremity and for fracture patterns typical of high-energy forces. PMID- 8229376 TI - The computerized tomography subchondral arc: a new method of assessing acetabular articular continuity after fracture (a preliminary report). AB - The criteria to treat acetabular fractures operatively versus nonoperatively continue to evolve. The technique of roof arc measurements was developed to identify the extent of superior acetabulum left intact after fracture. Computerized tomography (CT) of the superior 10 mm of the acetabular articular surface evaluates the area equivalent to roof arc measurements of 45 degrees. CT provides increased detail of the superior acetabulum involved with the fracture. Our current criteria for selecting cases for nonoperative treatment are as follows: 1. The acetabular articular surface is intact in the superior 10 mm of the joint on CT evaluation. (Fractures that enter the acetabular fossa, but not the articular surface in the superior 10 mm are included in this group.) 2. The femoral head remains congruent with the superior acetabulum out of traction on the anteroposterior and 45 degree oblique radiographic views of the pelvis. 3. When a posterior wall fracture is part of the injury pattern, a minimum of 50% of the posterior wall articular surface is intact at the most involved level as determined by CT. PMID- 8229377 TI - Intracompartmental pressure measurements in the normal forearm. AB - In 20 normal subjects, intracompartmental pressure measurements were made at three different sites in the volar forearm: half the distance between the medial epicondyle and ulnar styloid and at points 4 cm proximal and 4 cm distal. The pressure measurements were made using a hand-held digital compartment pressure monitor. The study demonstrated that in the uninjured volar compartment, clinically significant (5 mm Hg) intracompartmental pressure differences exist over distances as little as 4 cm. PMID- 8229378 TI - Fracture blisters: clinical and pathological aspects. AB - Fracture blisters are tense vesicles or bullae that arise on markedly swollen skin directly overlying a fracture. There is very little objective data in the literature detailing their characteristics and management. All fracture blisters that occurred over a 3 1/2-year period were studied retrospectively at four hospitals, of which three were level I trauma centers. A total of 53 blisters developed in 51 patients. They occurred in characteristic locations along the human musculoskeleton, most commonly overlying the tibia, ankle, and elbow. They arose within 24-48 h of acute injury in most instances. The timing of surgical intervention affected the occurrence of fracture blisters. Those patients with acute fractures who underwent open reduction internal fixation (ORIF) within 24 h of injury had the lowest incidence of fracture blisters (2.0%) compared with those delayed for > 24 h (8.0%) (p < 0.001). In those patients with fracture blisters present at time of surgery, patient care was affected in 10 of 13 cases (71%). Two of these were major complications occurring as postoperative wound infections. Other management problems consisted of delaying surgery, and changing in the operative plan. There were no adverse affects on patient care when the fracture blister developed postoperatively. Twenty-one fractures with blisters were treated by closed means, with the presence of fractures blisters delaying closed reduction and casting in two. Biopsy examination of 15 blisters supported the clinical impression that fracture blisters are subepidermal vesicles. The blister fluid was found to be a sterile transudate. Microbial evaluation of 11 ruptured fracture blisters demonstrated colonization (primarily with skin pathogens), occurred soon after blister rupture, and continued until reepithelialization. PMID- 8229379 TI - Elderly patients with hip fractures: improved outcome with the use of care maps with high-quality medical and nursing protocols. AB - In a prospective cohort study, 51 patients were treated with standard nursing and medical treatment after sustaining a hip fracture. A second group of 55 patients was treated with high-quality medical and nursing protocols outlined on our care map. The patient groups showed no significant differences preoperatively in terms of the important variables of age, mental status, marital status, accommodation, ambulation, fracture type or fracture treatment. Postoperatively the patients were followed for a 6-month outcome. The outcome of return to their place of accommodation and their previous level of function was considered grade 1. In grade 2, the patients lost one level of function or one level of accommodation. In grade 3 they lost one level of accommodation and one level of function. In grade 4 they were dead. Overall we were able to show that the patients in the study group had a statistically significantly better outcome (p = 0.036). In addition, they had significantly fewer postoperative complications (p = 0.01) and a significantly greater number of the patients returned home within 14 days of their admission. We feel that the medical and nursing protocols outlined are generally applicable and could significantly improve the outcome overall for elderly patients with fractured hips. PMID- 8229380 TI - Healing complications after internal fixation of trochanteric hip fractures: the prognostic value of osteoporosis. AB - In a 3-year prospective randomized trial, the prognostic value of osteoporosis in terms of predicting healing complications due to implant failures or loss of fracture reduction has been studied in 113 patients with inter-trochanteric hip fractures treated with either a sliding screwplate or Ender nailing. According to the Barnet and Nordin's osteoporosis index, 66 patients were considered to have moderate or severe osteoporosis. There were no differences between patients with or without osteoporosis as regards the number of fractures with collapse in varus > 10 degrees, redislocation > 10 mm, or compression exceeding 10 mm. Six months after surgery, 48 patients failed in healing or healed with complications. These cases disclosed increased osteoporosis in comparison with those showing noncomplicated healing. Patients with osteoporosis who had unstable fractures treated by Ender nailing were found to have the worst prognosis for healing complications (p = 0.006). Of the 33 fractures with implant failures, 25 (76%) occurred in patients with osteoporosis (p = 0.04). In patients with osteoporosis, the frequency of mechanical failures was significantly higher in fractures with the device positioned incorrectly. Independently of the device used, patients with unstable trochanteric hip fractures and osteoporotic bone were the group with the highest risk for failures of the implant. PMID- 8229381 TI - Clandestine femoral neck fractures with ipsilateral diaphyseal fractures. AB - We present a protocol for diagnosis of all femoral neck fractures associated with ipsilateral femoral diaphyseal fractures. A 30% incidence of delayed diagnosis has been reported by other investigators. Between 1982 and 1990, we have treated 32 patients with ipsilateral femoral neck and shaft fractures due to blunt trauma. Only 22 femoral neck fractures were diagnosed on prediaphyseal fixation radiographs. This left the 10 patients in this study who had normal prediaphyseal fixation radiographs and were subsequently found to have femoral neck fractures. The ipsilateral femoral neck fractures were found through a retrospective chart and radiographic review of all 555 femoral diaphyseal fractures identified through our trauma and fracture registries. The clinical and radiologic techniques for diagnosing the femoral neck fractures were presented. The time to union of the femoral shaft and neck was determined, and a preliminary radiologic assessment of the vascularity of the femoral head was made. Ten femoral neck fractures (31%) with normal preoperative radiographs were diagnosed after femoral diaphyseal fixation. One patient did not have a post-diaphyseal fixation radiograph. An incidental radiograph at 6 weeks showed a mildly displaced femoral neck fracture in an asymptomatic patient. At 16 weeks the patient became symptomatic, and a repeat radiograph showed the fracture. Five fractures were diagnosed in asymptomatic patients on routine post-femoral fixation radiographs. Two patients had normal post-femoral fixation radiographs, became symptomatic, and had their femoral neck fractures diagnosed on repeat radiographs at 3 and 7 days. One patient had normal pre- and postfixation radiographs, and on a 25-day routine femoral radiograph, the femoral neck fracture was diagnosed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229382 TI - The holding strength of cannulated screws compared with solid core screws in cortical and cancellous bone. AB - The comparative holding strength of cannulated screws (CS) versus solid core screws (SCS) has not been reported, although differences exist in the respective diameters of their outer thread and pilot drill holes. Our objectives were to characterize these differences and determine the holding power of CS compared with SCS in cortical and cancellous bone. The dimensions of the Synthes (Paoli, PA) 3.5-mm SCS, 3.5-mm CS, 6.5-mm SCS, and 7.0-mm CS were measured, and the cross sectional area for thread purchase was calculated. Using adult canine femurs, small-fragment 3.5-mm SCS were inserted in cortical (midshaft) and cancellous (condyle) bone of one limb, and CS were placed in similar locations in the contralateral limb. The same technique was used for large-fragment CS and SCS. Pull-out testing was performed using an MTS machine (MTS Systems, Minneapolis, MN) with axial loads applied at 5 mm/s, and data were analyzed to determine the effects of screw type, location, and size. Differences in CS versus SCS design result in higher cross-sectional areas available for SCS thread purchase. Yet no significant differences exists between screw types (SCS vs. CS) in either cortical or cancellous bone. In cancellous bone, large-fragment screws required more force to pull out than did small screws (p = 0.000). The mean force required to pull out small-fragment screws was higher in cortical bone than in cancellous bone (p = 0.000). These data suggest that the clinical decision to use CS versus SCS should not be based on pull-out strength. PMID- 8229383 TI - An experimental model of femoral condylar defect leading to osteoarthrosis. AB - The acute treatment of articular step-off injuries is based largely on reduction criteria, because the presence of residual incongruity has been correlated with the development of posttraumatic arthrosis (PTA). However, this association has not been demonstrated on a prospective basis. Using the rabbit femoral condyle, we developed a surgical model of articular condylar defect without sacrificing the axial alignment or inherent stability of the knee joint. Twenty weeks after the creation of 5-mm femoral condylar defects, progressive osteoarthritic changes were confirmed by radiographic, histological, and biochemical parameters. Osteophytes were observed on the medial aspect of operated knee joints in 67% of cases. Femoral and tibial articular cartilage at the site of the condylar defect exhibited fibrillation, hypocellularity, and severe loss of safranin-O staining. Focal areas of cartilage were denuded or replaced by pannus. In no case was femoral congruity restored by cartilage repair. Statistically significant decreases in proteoglycan content were demonstrated for cartilage sampled from the weight-bearing region of the condylar defect and from the tibial surface directly beneath it. These changes resemble those arising from previously reported models of osteoarthrosis. We present the model as a valid tool for the study of articular condylar defect and its role in the development of PTA. PMID- 8229384 TI - Segmental tibial shaft fractures treated with interlocking nailing. AB - Thirty-eight segmental tibial shaft fractures were treated with closed Grosse Kempf interlocking nails and retrospectively followed up for > or = 1 year (average 32 months). There was a 97% union rate and a 4.5 +/- 1.6 month union period. Knee as well as ankle range of motion was satisfactory. Significant complications included one aseptic nonunion (3%), which healed after the static locked nail was dynamized. We recommend that whenever possible, closed interlocking nailing should be used to treat closed or mild open (wound size < 1 cm) segmental tibial shaft fractures. PMID- 8229385 TI - The incidence of osteitis in open fractures: an analysis of 948 open fractures (a review of the Hannover experience). AB - Even though treatment protocols of open fractures have been improved in the past two decades, osteitis is still a major complication in these injuries. To investigate the primary factors responsible for posttraumatic osteitis, 19 cases of osteitis out of 297 open fractures (retrospective series from 1981 to 1983) and nine cases of osteitis out of 651 open fractures (prospective series from 1984 to 1989) were analyzed. The Hannover fracture scale was used for quantitative evaluation of the injury. A high prognostic index for bone infections was found for the amount of bone loss, the fracture type, the type of bacteriologic contamination, deep soft-tissue defects, compartment syndromes, vascular injuries, and soft-tissue infections. PMID- 8229386 TI - Distal locking screw insertion using a cannulated drill bit: technical note. PMID- 8229387 TI - Addressing the problems of minorities in oral and maxillofacial surgery. PMID- 8229388 TI - Treatment of fractures of the edentulous mandible, 1943 to 1993: a review of the literature. PMID- 8229389 TI - Use of homologous bone in maxillofacial surgery. AB - Homologous bone grafts were used in 135 maxillofacial surgical procedures, including acute midfacial fracture repair (n = 77), elective osteotomies of the facial bones (n = 35), secondary correction of traumatic deformities (n = 6), mandibular reconstruction (n = 10), facial bone augmentation (n = 5), and reconstruction of maxillary tumor defects (n = 2). Postsurgical complications occurred in five of the patients. This article reviews the rationale for using homologous bone grafts, their immune response, how they heal, and the risk of transmission of disease. PMID- 8229390 TI - Timing and dosage of postoperative radiotherapy for squamous cell carcinoma of the upper aerodigestive tract. AB - Seventy patients who received postoperative irradiation (PI) after curative surgery for stage III or IV squamous cell carcinoma of the upper aerodigestive tract were studied retrospectively to compare the rate of local and regional recurrence (LRR) and the effect of total dose on LRR rate in patients irradiated timely (n = 40) with those who were not (n = 30). Overall, the LRR rate was higher when PI was delayed than when timely (37% vs 20%). No advantage resulted from increasing total dose when PI was delayed; the LRR rate in the primary site and upper neck was 18% with less than 60 Gy and was 26% with 60 Gy or more; the LRR rate in the lower neck was 13% with 50 Gy or less and was 14% with more than 50 Gy. These data seem to corroborate the findings of other investigators regarding the importance of initiating timely PI, but not the observation that a pronounced delay is not detrimental provided higher tumoricidal doses are used. PMID- 8229391 TI - Augmentation of the maxillary sinus floor with autogenous bone for the placement of endosseous implants: a preliminary report. AB - Placement of endosseous implants in the atrophic maxilla is often restricted because of lack of supporting bone. In this article, experience with augmentation of the maxillary sinus floor with autogenous bone grafts to enable insertion of endosseous implants is described. The technique is aimed at providing a cortical layer on top of the graft to ensure a reliable seal of the maxillary sinus and to achieve optimal stability of the bone graft in case of simultaneously placement of dental implants. The procedure was used in 25 patients, using iliac crest grafts (22 patients, 86 implants), symphyseal bone grafts (two patients, six implants), or a maxillary tuberosity bone graft (one patient, one implant). Ninety-three Branemark implants (Nobelpharma, Gotenburg, Sweden) were inserted in 47 grafted maxillary sinuses. The mean follow-up was 16 months (range, 6 to 36 months). No inflammation of the bone grafts nor of the maxillary sinus occurred. The sinus membrane was perforated accidentally in eight cases during the surgical procedure. Five implants (5.4%), all inserted in iliac crest grafts, were lost during the healing period. The patients received implant supported overdentures (16 patients) or bone-anchored bridges (nine patients). From this preliminary study it is concluded that augmentation of the maxillary sinus floor with bone grafts for the insertion of endosseous implants is a promising solution for patients with atrophic maxillae and functional problems with their partial or full dentures. PMID- 8229392 TI - The fixed mandibular implant: a 6-year review. AB - In a review of 102 consecutive cases of the fixed mandibular implant there were no failures; all appeared clinically to be osseointegrated and functioning well. There were an unexpected number of gingival infections and instances of inflammatory gingival hyperplasia, but these have been reversible and manageable. A change in sleeve nut design has improved gingival health. The Fixed Mandibular Implant (Hall Reconstructive Systems, Carpinteria, CA) offers immediate denture stability and function, and provides the opportunity for a fixed, fixed removable, or overdenture prosthetic reconstruction. PMID- 8229393 TI - Prevention of postsurgical bleeding in oral surgery using tranexamic acid without dose modification of oral anticoagulants. AB - The hemostatic effect of tranexamic acid solution (4.8%) used as a mouthwash was compared with a placebo solution in 93 patients on continuous, unchanged, oral anticoagulant treatment undergoing oral surgery. The investigation was a randomized, double-blind, placebo-controlled, multicenter study. Before suturing, the surgically treated region was irrigated with 10 mL of tranexamic acid (46 patients) or placebo (47 patients) solution. The patients then performed mouthwash with 10 mL of the solution for 2 minutes four times daily for 7 days. The treatment groups were comparable regarding age, smoking habits, and surgery. Laboratory variables measuring vitamin K-dependent coagulation factors were within therapeutic ranges (international normalized ratio 4.00 to 2.10). One of the clinics used a different method for these measurements and therefore the levels of coagulation factor X in plasma obtained for the three clinics were compared. No significant difference in the range at which surgery was performed was found between clinics. In the placebo group, 10 patients developed bleeding requiring treatment, while none of the patients treated with tranexamic acid solution had bleeding. This difference was statistically significant (P < .01). The treatment with mouthwash was well tolerated. It was concluded that patients on oral anticoagulants can undergo oral surgery within the therapeutic range without reducing the dosage of anticoagulants, provided that local antifibrinolytic treatment with tranexamic acid solution is instituted. PMID- 8229394 TI - A comparison of microscrew and suture fixation for porous high-density polyethylene orbital floor implants. AB - Twenty oral and maxillofacial surgeons compared the surgical features of microscrew fixation technique with a suture technique for fixation of porous high density polyethylene orbital floor implants. One investigator then compared material features of these two fixation techniques using recently procured bovine ribs. The microscrew technique was found to be quicker (P > .01) and easier (P > .001), and it provided a higher quality of fixation (P > .001) than the suture technique. The microscrew/implant system had a greater pullout strength (P > .001) and shear strength (P > .001) than the suture/implant system. PMID- 8229395 TI - Changes in temporomandibular joint after mandibular subcondylar osteotomy: an experimental study in rats. AB - The objective of this study was to analyze the changes in the different components of the temporomandibular joint and their relation with age after subcondylar osteotomy. For this purpose 149 Wistar rats were divided into three groups: osteotomy, sham operation, and control. Two experimental ages (30 and 70 days) and radiologic, morphometric, macroscopic, and histologic methods of analysis were used. Different changes were observed in young and adult animals. Subcondylar mandibular osteotomy in growing rats caused anteroinferior displacement of the temporal and mandibular component of the joint. At the end of the experimental period only condylar displacement was maintained. Other changes were flattening of the mandibular condyle, which was transient, and lateral thickening of the articular disc. Subcondylar mandibular osteotomy in adult rats caused slight anterior displacement of the joint components at the end of the experimental period. At this age, although changes similar to those in growing animals were observed in some cases, in other cases the presence of pathologic findings, such as deformation of the condylar cartilage, thickening of the disc, intra-articular adherences, and osteolysis of the temporal fossa, were observed. Changes in the joint components were more marked in growing rats than in adults. In growing rats they affected the form of the condylar process to a greater extent, but the majority of these changes were transient. In adult rats, alterations in the joints were less pronounced, but they affected a greater number of joint components and were permanent. PMID- 8229396 TI - Fetal bone gap healing in utero. AB - The healing of fetal tibial bone after osteotomy with and without stable fixation has been reported previously. The present study was designed to evaluate fetal bone gap healing using a tibial ostectomy model in fetal sheep. Eighteen time dated pregnant ewes (20 fetuses, 34 experimental hind limbs) underwent intrauterine surgery at 95 days gestation (term, 145 days). A titanium miniplate was applied to the anterior aspect of the tibia and a longitudinal length of bone approximately 1.5 times its diameter was removed and the incision closed. The pregnancies were then allowed to progress until the ewe was killed at postoperative weeks 1, 2, 4, or 7. Assessments at that time included evaluation of gross morphology, histologic and radiologic appearance, and collagen analysis and hydroxyproline determination of the tissue within and at the borders of the gap. At 7 weeks, seven of nine bone gap specimens exhibited radiographic and histologic evidence of union with woven and lamellar bone. Hydroxyproline concentrations gradually increased within the bone gap over the period of the study. At all intervals, type I collagen composed over 90% of the collagen within the healing bony gap. Histologically and biochemically, the process appears to be similar to postnatal bone healing, albeit occurring at an accelerated rate. PMID- 8229397 TI - An animal model for subperiosteal tissue expansion. AB - An animal model for subperiosteal tissue expansion is described. Small tissue expanders were placed bilaterally and inflated in the buccal subperiosteal pockets of the mandible. Additionally, subcutaneous expansion was performed in the breast region. Twenty-one dogs were used in three experimental groups to evaluate the optimal size of an expander for this model, the optimal inflation regimen, and the histologic changes following subperiosteal expansion. Hemicylindrical expanders, 20 mm in width, 10 mm in height, and 30 mm in length, inflated with 1 mL saline every four days, were used for this model. It was found that the periosteum was replaced by fibrous connective tissue during expansion. Capsule formation following subperiosteal expansion was much more rapid than that following subcutaneous expansion. A greater increase in vascularity was noted in the subperiosteal expansion group and the subperiosteal control group than in the subcutaneous expansion group. In the subperiosteal expansion group, a thick fibrous capsule, with minimal inflammatory response, was observed 1 week after full inflation. Leaving the fully inflated subperiosteal expanders in place more than 1 month accelerated the resorption of the underlying bone. These results support the previously reported empirical regimens of subperiosteal tissue expansion. Furthermore, the excellence of the subperiosteal expanded bed as a recipient site for onlay grafting was confirmed. PMID- 8229398 TI - Research in oral and maxillofacial surgery, 1943 to 1993. PMID- 8229399 TI - Erosion of a Teflon-Proplast implant into the middle cranial fossa. PMID- 8229400 TI - Temporomandibular joint clicking only on closure: report of a case and explanation of the cause. PMID- 8229401 TI - Surgical treatment of Robin sequence and sleep apnea syndrome: case report and review of the literature. PMID- 8229402 TI - Non-Hodgkin's lymphoma presenting as submandibular swelling and palatal ulceration. PMID- 8229403 TI - The use of mandibular buccal cortical grafts in bimaxillary surgery. PMID- 8229404 TI - Avoiding fracture of titanium screws. PMID- 8229405 TI - Avoiding fracture of titanium screws. PMID- 8229406 TI - Assuming our responsibilities. PMID- 8229407 TI - Osteomyelitis of the jaws: a 50-year perspective. AB - The incidence of osteomyelitis of the jaws has decreased dramatically, except for a few subsets of individuals. This has been due, in no small part, to the availability of bacteriocidal antimicrobial therapy. The pathogenesis of osteomyelitis of the jaws is predominately due to odontogenic microorganisms rather than the classic skin contaminant, Staphylococcus. This causative relationship relegates the classification of osteomyelitis of the bimaxillary skeleton to predominately that of contiguous foci. These may be regionally progressive, secondary to microvascular compromise brought about by inherent flaws in regional anatomic calcified tissue vascular perfusion as well as by inflammatory metaplastic processes. Diagnosis is based on the presence of painful sequestra and suppurative areas of tooth-bearing jaw bone unresponsive to debridement and conservative therapy. This is usually accompanied by regional or systemic compromise of the immune response, microvascular decompensation, or both. Treatment of both acute and chronic forms of the disease, as outlined in Table 5, is successful if surgically supported. Sustained bacteriocidal antibiotic therapy is pertinent, especially in the face of potentially refractory virulent microorganisms and compromised regional vascular penetrance. The use of adjunctive hyperbaric oxygen therapy also may be included in the more refractory forms of osteomyelitis of the jaws to enhance the local and regional immune response of the jaws as well as to produce microvascular neoangiogenesis for reperfusion support. With resolution of infection, hard and soft tissue reconstruction may be necessary to augment the reparative process. PMID- 8229408 TI - Pediatric condylar fractures: a long-term follow-up study of 55 patients. AB - Fifty-five patients between 5 and 20 years old who were previously treated conservatively for a fracture of the mandibular condyle were recalled for follow up examination with a mean postoperative observation time of 10.1 years. Anamnestic information was obtained from a questionnaire, and clinical and radiologic examinations were performed. Anamnestic and clinical dysfunction indices were obtained. When the patients were divided into four different age groups it was found that the dysfunction index values increased significantly with increasing age at the time of trauma. Radiologic abnormalities such as reduced ramus height, deviation of the mandibular midline, and irregular shape of the condyle were seen frequently, but could not be correlated with the severity of dysfunction. No cases of ankylosis or serious asymmetry were found. The results support the opinion that conservative treatment is sufficient in pediatric patients, but in older age groups the results of conservative treatment are less satisfactory. PMID- 8229409 TI - Electromyographic evaluation of continuous passive motion versus manual rehabilitation of the temporomandibular joint. AB - There have been a myriad of rehabilitation techniques for patients after temporomandibular joint (TMJ) surgery, varying from professionally delivered physical therapy to self use of tongue blades. Most recently continuous passive motion (CPM) has been introduced for postsurgical TMJ patients. The benefits of CPM have been attributed both to its continuous and its passive nature. While the continuous nature of the therapy is accepted, questions remain regarding the passivity of the motion. The purpose of this study was to electromyographically evaluate the degree of activity that occurs with CPM compared with simple mandibular exercises in TMJ rehabilitation. Eight patients who had arthroscopy or arthrotomy were begun on a CPM device postsurgically. Electromyographic data were obtained for jaw closing and opening activity. Patients were measured using CPM as well as tongue depressors. Three different exercises were performed: 1) activity at rest was compared with rest with CPM; 2) opening and closing with CPM versus use of tongue depressors was compared; and 3) maximum opening to a comfortable position was compared using tongue depressors versus the CPM device in a fixed open position. The degree of opening was matched for both groups. A significant difference in electromyographic activity was shown between patients using CPM versus tongue depressors during the open/closing cycles. This suggests that TMJ CPM is truly passive. PMID- 8229410 TI - Evaluation of rigid internal fixation of mandible fractures performed in the teaching laboratory. AB - Rigid internal fixation of mandible fractures is advocated as an effective means of providing undisturbed healing and immediate function. However, its application in the clinical setting has resulted in many technique-related failures. To determine the reasons for clinical failure and to help develop improved means of providing successful fixation, the results of plate application in the teaching laboratory were evaluated. Seventy-four surgeons taking the AO/Association for the Study of Internal Fixation (AO/ASIF) maxillofacial course in Davos, Switzerland and 56 surgeons taking the advanced AO/ASIF course in Naples, FL applied rigid internal fixation devices to a mandibular fracture model. Treatment was evaluated for anatomic morbidity and effectiveness. In Davos, 92 of 121 subcondylar, body, symphysis, and ramus fractures (76%) were treated effectively. This included 22 of 34 symphysis fractures (65%). In Naples, 11 of 28 symphyseal fractures (39%) were fixed effectively with lag screws without anatomic morbidity. Technique failure is a frequent event in the application of rigid internal fixation devices to mandible fractures. Lag screw fixation of symphyseal fractures had a particularly high rate of technique-related failure. PMID- 8229411 TI - Rigid fixation of comminuted mandibular fractures. AB - Treatment of comminuted mandibular fractures has traditionally involved closed reduction with external fixation in an effort to avoid stripping periosteum from the bony segments. The purpose of this study was to evaluate retrospectively the success rate of rigid fixation used to treat 16 consecutive comminuted fractures of the mandible in 15 patients. The comminuted fractures were as follows: symphysis and body, 10; angle, 3; and ramus, 3. In 13 cases, AO stainless steel reconstruction plates were applied, with a minimum of three 2.7-mm tapped bone screws in each of the two stable segments. In most cases additional screws were placed into the comminuted fragments to stabilize them. In two patients, the mandibular ramus was comminuted (one case bilaterally), which made it impossible to place a large plate. These fractures were stabilized with multiple titanium miniplates (2.0 mm, tapped). All patients' fractures healed to a bony union without bone grafting. The mean maximum incisal opening at longest follow-up was 40 mm (range, 20 to 50 mm). All patients had a satisfactory facial form and none required further surgery for facial recontouring or malocclusion. Complications were observed in three patients. Two patients (13%) developed infections. Both infections were caused by loose hardware and responded to removal of the hardware and did not require any other treatment. One mandible refractured during manipulation after coronoidectomy to treat mandibular hypomobility, a sequellae of a gunshot wound that traversed both mandibular rami. This fracture healed after plate removal and a course of maxillomandibular fixation without bone grafting. This study suggests that rigid fixation of comminuted mandibular fractures is a viable treatment that satisfies the necessity for reestablishment of form and function with minimal morbidity. PMID- 8229412 TI - Mandibular reconstruction using hydroxylapatite granules, autogenous bone, and a cervical island skin flap. AB - Ten patients with gingival carcinoma who were treated by marginal mandibulectomy were reconstructed using hydroxylapatite granules, autogenous bone chips, and a cervical island skin flap. The cervical flap was used to produce a cover and partition, designed to prevent the displacement of the mixture of hydroxylapatite and bone. Reconstruction was successfully achieved in all patients. Loss of height of the reconstructed mandible over an average 18-month follow-up ranged from 8% to 22%. One patient has worn a full denture and five patients wear partial dentures. PMID- 8229413 TI - Metastatic tumors to postextraction sites. AB - Review of the literature revealed 55 cases where tooth extraction preceded the discovery of metastases. The lung and breast were the most common sources of the metastasis, and the mandibular premolar area was the most common site. A soft tissue mass extruding from a recent extraction wound, and accompanied by pain, were the main symptoms in most patients. The mean time from discovery of the metastasis to death was 3.8 months. Tooth extraction appears to serve as a promoting factor in the metastatic process. A case of metastatic transitional cell carcinoma of the urinary bladder involving the area of a recently extracted mandibular third molar is reported. PMID- 8229414 TI - Long-term observation of the relation between pain intensity and serum carbamazepine concentration in elderly patients with trigeminal neuralgia. AB - The relation between changes in pain intensity and serum carbamazepine (CBZ) concentration was studied in five elderly patients with trigeminal neuralgia. Measurement of pain intensity and serum CBZ concentration, and blood studies, were performed approximately every 2 or 3 months after initiation of CBZ therapy. Fluctuations in pain intensity were observed in four patients. In these patients, even when the serum CBZ concentration was maintained within a relatively high range, there were periods in which pain intensity increased. On the other hand, there were periods in which pain was relatively well controlled even though the CBZ concentration was relatively low. There were considerable interindividual differences in response to CBZ. The fluctuations in pain showed that the CBZ response curve varies from period to period even within an individual. It was concluded that it is essential to observe spontaneous variations in pain intensity and monitor serum CBZ during long-term administration. PMID- 8229415 TI - Osteoinductive, morphologic, and biomechanical properties of autolyzed, antigen extracted, allogeneic human bone. AB - Autolyzed, antigen-extracted, allogeneic (AAA) bone was prepared from human cortical bone and its morphologic, biomechanical, and osteoinductive properties were compared with untreated (frozen) as well as lyophilized human bone. Scanning electron microscopy revealed removal of inorganic calcium phosphates and persistence of shrunken collagen fibrils on the surface of AAA bone matrix. Biomechanical testing of differently prepared bone samples showed that lyophilization increased both the modulus of elasticity (P < .00001) and the compressive strength (P < .00001). Depending on the depth of decalcification in the preparation of AAA bone, both measured values decreased in rehydrated AAA bone compared with untreated bone (P < .00001). Completely demineralized and rehydrated AAA bone was soft, flexible, and showed very little compressive strength. Differences in biomechanical behavior between samples drilled longitudinally or perpendicularly to the diaphyseal bone axis were observed. Xenogeneic human bone samples were implanted in muscle pouches of Sprague-Dawley rats for 6 weeks. AAA bone implants showed chondrogenesis and osteogenesis in 50% of the cases, while untreated or lyophilized bone implants induced no new cartilage or bone formation. As decalcification exposed xenogeneic organic matrix components, AAA bone implants provoked the highest inflammatory reaction. When AAA bone samples were implanted in immunosuppressed rats, the inflammatory reaction was suppressed and 94% of the implants showed endochondral bone formation. The chondroinductivity of the bone samples also was tested in vitro using neonatal rat muscle tissue to avoid interference with inflammatory cells and secreted cytokines. In this assay, 68% of AAA bone samples induced chondroneogenesis, while untreated as well as lyophilized bone samples failed to induce any cartilage formation. The results clearly demonstrate that AAA bone has osteoinductive properties. Biomechanical stability of AAA bone implants depends on the degree of demineralization. Thus, they can be prepared in an appropriate manner for different indications in oral and maxillofacial surgery. PMID- 8229416 TI - Development of chronic mandibular osteomyelitis in a miniswine model. AB - Previous attempts to develop a reproducible model of chronic mandibular osteomyelitis have met with limited success. In this study, osteomyelitis was produced in the mandibles of eight adult Yucatan miniswine by the intramedullary application of sodium morrhuate, Staphylococcus aureus, and either polymethylmethacrylate bone cement or bone wax. At 8 weeks' postinfection, the mandibles were surgically debrided and specimens were obtained for culture. Although all of the animals developed clinical evidence of osteomyelitis that was supported by positive cultures, the original organism (S aureus) was recovered only from those animals where bone wax had been used to seal the cortical defects. This animal model may be useful for evaluating newer treatment modalities for chronic osteomyelitis. PMID- 8229417 TI - Evaluation of new high-performance calcium polyphosphate bioceramics as bone graft materials. AB - The purpose of this study was to evaluate the ability of a recently developed porous calcium polyphosphate bioceramic (CPB) to function as a bone graft substitute. After six weeks, postsurgical extraction of the mandibular first and second molars, alveolar ostectomies were performed bilaterally in five dogs. The ridge forms were then restored using the CPB implant material on one side and the autogenous bone obtained from the contralateral ostectomy site on the other. The graft and implant sites were retrieved after 4 months and decalcified and undecalcified sections were prepared for special staining (modified Attwood) and subsequent light microscopy and histomorphometry. In addition, the undecalcified sections were prepared for histometry using backscattered electron imaging (BSEI). Histologically, the CPB implants showed extensive vascularization and cellularity within an "invading" loose connective tissue matrix. On the opposite side, the loose connective tissue of the autografts showed hypovascularity and hypocellularity. Neither the implants nor the autografts showed any histologic evidence of an inflammatory reaction. Using light microscopic histomorphometry, the implants showed a higher incidence of union than the autografts. On BSEI histometry, the CPB implants showed significantly greater new bone formation than the autografts. This study reveals that porous CPB possesses certain characteristics essential for the "ideal" implantable bone substitute necessary for the repair of craniofacial and other bony defects. PMID- 8229418 TI - Tissue banking safety: caveats and precautions for the oral and maxillofacial surgeon. AB - Oral and maxillofacial reconstructive surgeons using allogeneic tissues have expressed justifiable concern over the safety of these tissues as they relate to the transmission of infectious disease. This report reviews cases of infectious disease transmission from inadequately screened donors of allogeneic tissues, as well as those related to improper sterilization and cataloging of these tissues. It is concluded that good judgment and attention to good science on the part of the tissue bank as well as the surgeon can maximize the ability to place contamination-free specimens, thereby avoiding complications similar to those described. PMID- 8229419 TI - Changing distribution of oral and maxillofacial surgeons: 1982-1991. AB - This article reviews the 1991 and past two American Dental Association studies on the distribution of dentists, providing specific information on the evolving numbers of oral and maxillofacial surgeons at the national, regional, and state levels. PMID- 8229420 TI - A mass in the floor of the mouth. PMID- 8229421 TI - Joint infection as a complication of temporomandibular joint arthroscopy: a case report. PMID- 8229422 TI - Hyperplasia of a costochondral graft in an adult. PMID- 8229423 TI - Removal of a maxillary third molar from the infratemporal fossa by a temporal approach and the aid of image-intensifying cineradiography. PMID- 8229424 TI - Reconstruction of the lower lip with an upper lip island flap. PMID- 8229425 TI - A refined technique for oral commissurotomy. PMID- 8229426 TI - A modification of the interim splint for large maxillomandibular discrepancies. PMID- 8229427 TI - Injury to tooth buds from rigid fixation screws. PMID- 8229428 TI - Is the mandibular condyle necessary? PMID- 8229429 TI - Differences among intravenous extravasations using four common solutions. AB - A frequent complication of intravenous therapy is extravasation (infiltration) of the infused fluid into the interstitial tissues. This study compares infiltrates intentionally made using different IV solutions regarding surface assessment and the volume of infiltrate as quantified by magnetic resonance imaging. Solutions differed significantly concerning pain, surface area of induration, and volume at the site of infiltration. PMID- 8229430 TI - Treatment of infusion-related phlebitis: review and nursing protocol. AB - Infusion-related phlebitis is a frequently occurring, but often preventable complication associated with intravenous therapy. Early detection with prompt, consistent nursing intervention decreases the occurrence and severity of cannula related phlebitis. This article reviews the clinical aspects of phlebitis and describes a comprehensive approach for the treatment of infusion-related phlebitis. PMID- 8229431 TI - An innovative preceptor program for intravenous home care nursing. AB - The demands of intravenous home care nursing require a comprehensive orientation of newly hired staff. This article describes a model preceptor development program to promote collaboration in the orientation experience. Nurses are selected by their managers to attend a 3-day training program. An evaluation questionnaire is given to participants at the end of the program. Results indicate that at the conclusion of the program, the attendees feel prepared to assess learning needs, to plan instructional activities, and to evaluate learning outcomes of their future preceptees. This program can easily be adapted for use by other home care agencies providing IV nursing. PMID- 8229432 TI - Competency for intravenous nursing practice. AB - A definition of intravenous nursing competency is performing IV therapy in an exact and effective manner using the appropriate nursing, technical, and specialized skills. A competency-based evaluation tool has been developed to establish guidelines for orientation and evaluation in coordination with the Intravenous Nursing Standards of Practice, published by the Intravenous Nurses Society (1990). PMID- 8229433 TI - A process for selecting a safety needle system in a community teaching hospital. AB - Healthcare workers, especially intravenous nurses, are wary of the threat of bloodborne pathogens. In light of the Bloodborne Pathogen Standard issued by the Occupational Safety and Health Administration (OSHA), healthcare institutions must examine occupational exposures to blood and other infectious materials and investigate ways to minimize risks. The author describes the experiences of a community teaching hospital and the progressive steps involved in selecting a safety needle system. PMID- 8229434 TI - Effectiveness of disinfectant techniques on intravenous tubing latex injection ports. AB - This randomized, experimental-design study compared the effectiveness of two different disinfectant techniques for eliminating bacteria from intravenous tubing latex injection ports. The two treatment groups received a one-minute preparatory scrub of 70% isopropyl alcohol or povidone-iodine; the control group did not receive any scrub. Results showed approximately equal effectiveness of 70% isopropyl alcohol or povidone iodine on the growth of all micro-organisms. The control group ports showed significant growth of all microbes studied. PMID- 8229435 TI - Shared vision. PMID- 8229436 TI - [Clinical views of cases with discontinuity of the ossicular chain]. AB - It is well known that reconstruction for discontinuity of the ossicular chain, without inflammatory disease, has a good prognosis. Eleven cases of congenital ossicular chain malformation without ossicular fixation and 6 cases of ossicular chain injury due to the head trauma or an earpick , were treated surgically. The method of reconstruction and the prognoses, according to short-term and long-term follow up, were studied in these 17 cases. The following results were obtained: 1) In almost all cases, the pure tone audiograms were flat or slightly rising types, and the mean air conductive hearing loss was about 60 dB. Only 2 cases showed the falling type with conductive hearing loss. Connective tissue was observed at the incudo-stapedial joint intraoperatively in these two cases. 2) Mean hearing improvement was in 31.8 dB in the low frequency region (125, 250, 500 Hz), 22.7 dB in the middle frequency region (500, 1000, 2000 Hz), and 12.9 dB in the high frequency region (2000, 4000, 8000 Hz), within 3 weeks after operation, in all 17 cases. 3) No clear changes in hearing level were seen postoperatively, for the four periods evaluated; 1-3 weeks, 1-3 months, 4-6 months and over 7 months in all cases. 4) The cases in whom the reconstruction was performed between the incus and stapes, especially the foot plate of the stapes, showed poor hearing recovery. 5) There was no relation between hearing recovery and the prostheses used in reconstructions. However, it was thought that pieces of the patient's own cartilage or bone should be used whenever possible. PMID- 8229437 TI - [Study on consonant confusion in Japanese patients with sensorineural hearing loss]. AB - The speech discrimination scores of a 57S word list were analyzed in 180 patients with sensorineural hearing loss. Confusion matrices were made on consonant groups of voiceless, voiced and nasal consonants. In succession, the incidence of consonants was compared between the 57S word list and Japanese conversation. Consonants with high incidence in Japanese conversation were t, h, d, g, m, and n. Patients with sensorineural hearing loss are apt to confuse t and h to k, d to b or r, g to b, d or r, m to n or r, and n to m or r. It is effective to correct these instances of consonant confusion, for improving conversational ability in Japanese patients with sensorineural hearing loss. PMID- 8229438 TI - [Magnetic stimulation of the facial nerve]. AB - Intracranial activation of the facial nerve and the face-associated motor cortex are now possible with noninvasive magnetic stimulation techniques. Compound muscle action potentials (CMAPs) and the Blink reflex, in response to magnetic stimulation, were investigated. Subjects were 10 normal controls and 2 Bell's palsy patients. CMAPs were elicited in the orbicularis oris muscle by magnetic stimulation at the parieto-occipital skull and stylomastoid foramen. Furthermore, CMAPs were evoked by a magnetic coil oriented over the cortex. CMAP recording was possible with magnetic stimulation and the latencies of CMAPs at the parieto occipital skull were slightly greater than those at the stylomastoid foramen. In 10 normal subjects, the mean onset latency following transcranial magnetic stimulation of the facial nerve at the parieto-occipital skull was 5.07 msec (SD = 0.40), while transcutaneous latency at the stylomastoid foramen was 2.77msec (SD = 0.539). In the blink reflex, R1 latency was 10.99 msec (SD = 1.27), ipsilateral-R2 latency was 37.46 msec (SD = 2.57), and contralateral-R2 latency was 38.925 msec (SD = 3.20). The blink reflex thus had a configuration similar to that evoked by conventional electrical stimulation. In the patients with Bell's palsy, CMAPs elicited by magnetic stimulation were of low amplitude with normal latency. However, in the blink reflex, only a contralateral R2 response could be recorded, and R1 and ipsilateral-R2 showed no response to stimulation at the affected side. Investigation of patients with Bell's palsy using this technique may therefore prove useful in the evaluation of peripheral facial nerve disorders. PMID- 8229439 TI - [Patient selection for adult cochlear implantation and its experience]. AB - From 1986 to 1992, 61 adults with bilateral profound hearing loss visited our hospital for assessment as candidates for cochlear implantation. The selection process consists of the following steps: 1) taking a clinical history, 2) general otological examinations, 3) pure tone audiometry 4) hearing aid evaluation, 5) promontory stimulation testing and 6) CT scans and MR Imaging of the temporal bone. We informed patients and their families of cochlear implantation as well. Consequently 7 out of 61 cases (11.7%) were selected and underwent multi-channel cochlear implantation. Twenty-eight cases were judged to have better hearing with a hearing aid because of a residual hearing. Twenty-one cases were considered to be inappropriate candidates after informing of the cochlear implant procedure. Needless to say, the promontory stimulation testing and imaging studies are significant in patient selection, however, there was only a few who were excluded by these examinations. PMID- 8229440 TI - [Evoked oto-acoustic emissions in patients with secretory otitis media]. AB - In many studies, Evoked Otoacoustic Emissions (EOAEs) have been routinely documented in subjects with normal middle ear function showing tympanogram (TG) A type. However, middle ear dysfunction may influence the measurements of EOAEs. We investigated the EOAEs in ears with secretory otitis media (SOM) in order to understand the effects of middle ear dysfunction on EOAE measurements. We present data from 73 SOM ears of 38 subjects aged 3 to 58 (mean = 9.4 years) and data from 61 ears of 54 subjects with sensorineural hearing loss as a control. Generally, the EOAE thresholds of ears with mild sensorineural hearing loss have a good correlation with the audiometric hearing threshold level. On the other hand, the EOAE thresholds of ears with SOM were on the average higher than those expected from the audiometric hearing threshold level. In this paper, we classified TG B type further into TG B1 type and TG B2 type according to our criteria. Though neither TG B1 type nor TG B2 type revealed peaks in the tympanogram (-300mmH2O < or = < or = 200mmH2O), the 2 groups differ in that TG B1 type seems to have its peak at pressures under -300mmH2O on tympanography while TG B2 type seems to have no peaks in that pressure range. As a consequence of this classification, most of the EOAE thresholds of SOM ears showing TG B2 type were scaled out (> or = 40 or 50 dB nHL).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229441 TI - [Cholesteatoma in children--sex differences]. AB - It has been reported that cholesteatoma is more aggressive in children than in adults and that it affects boys more often than girls. We found particularly prominent sex differences in the clinical characteristics and behavior of cholesteatoma in children, by comparing them with those of acute otitis media (AOM), otitis media with effusion (OME) and adult cholesteatoma. We retrospectively analyzed the clinical records of children diagnosed as having cholesteatoma by comparing their courses with those of AOM, OME, chronic otitis media, microtia and congenital malformation of the ossicles. The boy-dominant tendency were observed in OMA, OME, pediatric cholesteatoma, microtia and congenital malformation of the ossicles. Cholesteatoma has a tendency to fill the middle ear cleft in boys, in whom the most extensive cholesteatomas were observed. If OMA, OME, and cholesteatoma in children are considered to be a series of inflammatory middle ear diseases, a common factor must be involved in the boy-dominant tendency for this series of inflammatory middle ear diseases. We have proposed six factors contributing to sex differences in pediatric cholesteatoma. Among these, immunological, environmental and congenital factors were thought to be responsible for the sex differences. PMID- 8229442 TI - [Carcinoma of the nasopharynx: analysis of treatment results in 51 patients]. AB - The outcomes of 51 cases of nasopharyngeal carcinoma, treated in our hospital between July 1971 and June 1991, were evaluated. Factors which appeared to influence prognosis were assessed by the Kaplan-Meier method. The ages of the patients ranged from 18 to 74 years. There were 38 males and 13 females. The cumulative five-year survival rate for the whole study population was 60.6%. The UICC classification (1987) was used for disease staging. The percentages of patients surviving after five years were: 100% (n = 1) for stage II, 75% (n = 4) for stage III, and 58.1% (n = 46) stage IV. The histological type with the highest survival rate was poorly differentiated squamous cell carcinoma, which had a 69.8% (n = 37) survival rate. The anatomical sites of primary tumors were the lateral wall (62.4%, n = 33) and the posterior wall (54%, n = 33). Percentages of patients according to classification based on initial therapy were as follows: Combined radiotherapy and chemotherapy group, 61.5% (n = 22); radiotherapy alone group, 56.5% (n = 22). The incidences of local recurrence according to initial therapy were 36% in the combined therapy group and 50% for the radiotherapy alone group. There were no metastases to the cervical lymph nodes after initial therapy in stage II and III. In stage IV, the rate of metastasis was lower when combined therapy was given. When the relationship between initial therapy and distant metastasis was evaluated, the rate of metastasis was lower in the combined therapy group. We conclude that combined therapy is necessary in the treatment of nasopharyngeal carcinoma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229443 TI - [Skull base surgery using the degloving technique--an approach without facial scarring]. AB - Skull base surgery has been performed in 16 cases for en bloc resection of the nasal cavity and paranasal sinus carcinoma invading the anterior and middle skull base in our department. The surgical approach to skull base surgery is mainly a combined craniofacial approach, which includes craniotomy via a coronal incision intracranially and a transfacial or transoral approach extracranially. The direct transfacial approaches, such as lateral rhinotomy, Weber-Furguson incision, and midfacial split, resultin facial scarring. The midface degloving procedure was reported by Casson in 1974. This method can expose the midfacial skeleton by lifting the soft tissue from the face through an intraoral approach without external scarring. The authors have modified Casson's facial degloving technique and applied it to skull base surgery. The entire cranium and facial skeleton can be extensively degloved by using coronal, gingival and conjunctival incisions. Then, en bloc resection of the craniofacial compound segment can be accomplished through this degloving approach without external scarring of the face in anterior and middle skull base surgery. The authors have performed this procedure in 2 cases, and proved it to be an extremely valuable technique, displaying excellent cosmetic and functional results with minimal complications. PMID- 8229444 TI - [Clinical study of intractable recurrent ulcer of the oral cavity and pharynx]. AB - The etiology and clinical definition of intractable recurrent ulcer of the oral cavity and pharynx were long unknown. Then, intractable recurrent ulcer of the oral cavity and pharynx was defined as irregular ulcerative lesions occurring only in the oral cavity and pharynx, showing no specific findings in clinicohematological examinations, which recurred easily and were resistant to various treatments for at least one month. Twenty-five cases of intractable recurrent ulcer of the oral cavity and pharynx were treated in our hospital. Patient age, sex, chief complaint, past history, duration of illness, the locations and clinical findings of the ulcers, clinico-hematological examinations, the pathology of the ulcers, therapy and prognosis were analysed. Disease specificity was found in age, sex, past history, locations and clinical findings of the ulcers, and prognosis. Our results suggest that intractable recurrent ulcer of the oral cavity and pharynx is a characteristic disease. The possibility that HLA-linked genetic factors play a role in the development of intractable recurrent which ulcers, which in addition to other factors, induce local immunoreaction which then prolongs the course of this disease, was suggested. PMID- 8229445 TI - [A three-dimensional reconstruction of the temporal bone by the helical scanning CT and its clinical application]. AB - The current availability of 3 dimensional (3-D) imaging from Computed Tomography (CT) has yielded new anatomical information and pre-and postoperative evaluations. However, little discussion as to the 3-D structural image of the temporal bone has been reported because conventional CT does provide sufficient data to produce such images. The Helical scanning CT gathers continuous and multiple slice image data since it consists of an X-ray tube that continuously rotates around the patient while the patient moves continuously into the CT scanner. Thus, application of the Helical scanning CT has made it possible to reconstruct 3-D images of the minute and complicated structure of the temporal bone. We evaluated 3-D images from 9 typical cases, examined from February to October 1992. As a result, we found that the 3-D images reconstructed with this system are useful for evaluation of the postoperative state of tympanoplasty, the diagnosis of anomalies of the bony labyrinth, and examining the extent of bone destruction induced by trauma, cholesteatoma, etc. PMID- 8229446 TI - [Observation of calcification of thyroid tumors determined by CR and CT]. AB - Detection of calcification of the thyroid gland is very useful for the clinical diagnosis and therapy of thyroid cancer. In this study, we evaluated the usefulness of FCR (Fuji computed radiography) and CT by analyzing the correlation between the imaging findings of calcification and histological diagnosis. Fifty one cases of follicular adenoma and 63 cases of differentiated carcinoma underwent FCR and CT. All cases undergoing operation received histological examination. Calcification was detected in 19.6% of follicular adenomas and in 46.0% of differentiated carcinomas by FCR, and 37.3% of follicular adenomas and in 63.5% of differentiated carcinomas by CT. Among cases without calcification on FCR, CT detected calcification in 26.7%. These differences between the results of FCR and CT were found in cases with small calcifications, low thyroid gland position and in whom differentiation between tracheal cartilage calcification and thyroid carcinoma calcification was difficult. In these cases, calcification was difficult to detect by FCR alone. Patients with follicular adenoma aged > or = 60 years showed a higher prevalence of calcification than younger patients. In cases of differentiated carcinoma, no relationship was seen between age and the prevalence of calcification. The classification of calcification shape by FCR showed that the majority of psammoma bodies, as well as sharp and irregular shapes, were found in differentiated carcinoma while round calcifications were found in follicular adenomas. On CT, calcification inside the tumor was found in 73.7% of follicular adenomas and in 92.5% of differentiated carcinomas. FCR and CT are more useful for the detection of calcification than conventional radiography and are of great value in screening for thyroid tumors. PMID- 8229447 TI - [Relationships between ototoxicities and chemical structures of ototoxic drugs]. AB - The relationships between degrees of ototoxicity and the chemical structures of ototoxic drugs were investigated using isolated bullfrog semicircular canals. Thirteen derivatives of tuberactinomycin (Tum), a peptide antibiotic with pharmacological characteristics similar to those of kanamycin, were prepared by replacing the R1 branch with various amino acids. The degrees of ototoxicity were measured in these derivatives as well as in different kinds of aminoglycoside (AGs) antibiotics (TOB, GM, NTL, ISP, AMK). In order to measure the degree of ototoxicity, the ampullary nerve action potential of isolated bullfrog posterior semicircular canal, in response to mechanical endolymphatic flow, was recorded in Frog Ringer's solution and in the presence of different amounts of the various drugs. The degree of ototoxicity was determined by the amount of decrease in the maximal spike account. The degrees of ototoxicity of AGs were calculated to be in the order TOB > GM > ISP > AMK > NTL. This result was in agreement with those of previous morphological investigations. The derivatives of Tum with an R1 branch containing an acidic or a basic amino acid showed greater ototoxicities than those containing aliphatic side chains. The degree of ototoxicity of each drug tended to differ with each concentration, in both AG and Tum derivatives. Thus, the result indicate that chemical structure may be closely related to the degree of ototoxicity. PMID- 8229448 TI - [Clinical study on perforation of the tympanic membrane and discussion based on experimentally induced tympanic rupture]. AB - Traumatic perforation of the tympanic membrane (TM) was clinically studied by analyzing photographs of 231 ears treated in the Department of Otolaryngology of Tokyo Women's Medical College from May 1983 to December 1991. The dynamic features of TM and the mechanism of perforation are discussed. RESULTS AND DISCUSSION: 1) Direct and indirect injuries of the TM through the external auditory canals were observed in 101 and 130 ears, respectively. One hundred and twenty-seven injured ears were seen in males and 104 in females. Subjects ages ranged from 1 to 64 years. One hundred and thirty ears were on the left side and 101 on the right. 2) Approximately 80% of the perforated regions were observed in the antero-inferior or postero-inferior quadrant. "Triangular-type" perforation was predominant among direct injuries and "slit type" among indirect injuries. 3) Approximately 80% of the cases had a TM defect area smaller than 20%. 4) One hundred and twenty of 127 ears which were treated within 7 days after injury were healed by conservative treatment. One hundred and four ears among these 120 ears healed spontaneously. The average healing period for these 104 ears was 25 days. 5) It was suggested that the age of patients and the defect area of TM were factors that delayed healing. 6) The minimum breakage stress of human normal TM (the right angle to the radial fibers of TM) was 13.7 gf/mm2. 7) It was presumed that the strain stress and shear stress of TM contributed to traumatic TM perforation in indirect injuries. PMID- 8229449 TI - [Multiple primary cancers in patients with head and neck cancer and history of tobacco smoking and drinking]. AB - In a total of 2083 cases of hollow organ cancer of the head and neck region treated in our institute in the past 20 years, the relative risk of second primary cancers was statistically studied by the person-year approach. The organ association in multiple primary cancers and the relationship between tobacco and alcohol consumption and the development of second primary cancers were also reviewed. The incidence of second primary cancers was significantly (p < 0.05) higher in patients having a lesion at the tongue (n = 502), mesopharynx (n = 188), hypopharynx (n = 224) and larynx (n = 621) and in the oral cavity (n = 203) than in the general population. With regard to organ association in multiple primary cancers, cancers in the oral cavity, esophagus and at the pharynx were found to occur as second primary cancers of the tongue, oral cavity, meso hypopharynx and larynx at significantly high rates in males. Male laryngeal cancer was related to the lung. No relation of female lingual cancer to the esophagus was noted. The incidence of second primary cancer was very high in persons who had been heavy smokers and/or heavy drinkers before the occurrence of their primary cancer in the head and neck region. In these people, second primary cancers were found to occur at high rates as cancers related to tobacco smoking and drinking. Otherwise, in cases with head and neck cancers less related to tobacco smoking and drinking, the incidence of second primary cancers was low. From the above results, tobacco smoking and drinking may be closely related to a high incidence of second primary cancer following the first primary cancer in the head and neck region. PMID- 8229450 TI - Integrins and tumour progression. PMID- 8229451 TI - Ulcer-associated cell lineage ('pyloric metaplasia') in Crohn's disease: a lectin histochemical study. AB - Chronic intestinal ulceration in Crohn's disease is associated with the development of an epidermal growth factor-secreting cell lineage, or 'ulcer associated cell lineage' (UACL). Expression of oligosaccharides by UACL was studied using a panel of 25 biotinylated lectins with an avidin peroxidase revealing system and compared with that of adult and fetal Brunner's glands, gastric antral mucosa, and 'gastric metaplasia' within the duodenum, in order to clarify further the interrelationships of these lineages. UACL was obtained from ileal resections performed for Crohn's disease. Lectin binding of the glandular component of UACL closely resembled that of antral mucosal glands and also that of fetal and adult Brunner's glands. Lectin binding of the ductal component of immature UACL, in which a surface component had not developed, resembled that of the gland. The surface and ductal components of mature UACL showed a distinct lectin-binding profile, which was very different from that of the gland, but closely resembled that of antral foveolar epithelium and 'gastric metaplasia' within the duodenum. It is concluded that there is differentiation of UACL from the glandular to surface components and that oligosaccharide expression of the lineage reflects that of normal Brunner's gland and gastric antral mucosa. PMID- 8229452 TI - p53 antigen in cervical condylomata, intraepithelial neoplasia, and carcinoma: relationship to HPV infection and integration. AB - It has been proposed that wild-type p53 cell-regulating functions are annulled in human cervical carcinomas, either by mutations in the human papillomavirus (HPV) negative cases or as a consequence of their complexing with HPV E6. The aim of this study was to test this hypothesis on 39 fresh cervical biopsies by p53 immunocytochemistry (ICC) with antibody PAb 240 and with NISH (non-isotopic in situ hybridization) and PCR (polymerase chain reaction) for HPV detection. p53 protein was present in the basal layer of pure wart virus infection; the basal to middle third of CIN (cervical intraepithelial neoplasia); in 19/22 (86 per cent) HPV-positive cervical carcinomas, ten of which contained integrated HPV; and in 4/8 (50 per cent) HPV-negative cervical carcinomas. Dual detection of p53 antigen and HPV 16 DNA in the same sections demonstrated either p53 protein or integrated HPV 16 alone in the majority of cells. Co-localization of both signals was only evident in isolated cells. These data suggest that PAb 240 immunoreactivity is not mutant-specific. They are, however, consistent with the conformation hypothesis which proposes that wild-type p53 changes from a suppressor (PAb 240 negative) to a promoter (PAb 240-positive) form during cell growth response. Hence, according to this hypothesis, p53 protein expression may represent either the wild-type promoter form or mutant p53 protein, both of which share the same conformation. This may explain co-localization of p53 and HPV in some tumours. However, the absence of p53 protein in 50 per cent HPV-negative squamous cell carcinomas suggests that not all HPV-negative tumours accumulate p53 protein. PMID- 8229453 TI - The ulcer-associated cell lineage: the gastrointestinal repair kit? PMID- 8229454 TI - Papillomavirus screening in cervical cell samples using dual-label dot-blot analysis. AB - The presence of human papillomavirus (HPV) in cervical cells is closely related to the development of cervical carcinoma. Detection of virus may be by Southern blot, dot blot or the highly sensitive polymerase chain reaction. Whatever method is employed, there are problems of false negatives due to poor clinical samples in which the DNA may be degraded or is absent altogether. Here we describe a new method of dual labelling for dot blots using a 32P-labelled probe for HPV and a 35S-labelled probe for human actin genes. The samples were counted on a Beta plate flat-bed scintillation counter and the data analysed to separate the activities of the two isotopes. The counts from the actin probe show whether human DNA is present or not and false negatives from this cause may thereby be eliminated. The counts due to HPV when compared with those for actin give a quantitative measure of HPV abundance for the particular sample and this may have clinical relevance. PMID- 8229455 TI - Proliferative state of the urothelium with benign and atypical changes. Correlation with transferrin and epidermal growth factor receptors and blood group antigens. AB - We attempted to investigate how the proliferative state of the urothelium correlates with the reactivity for transferrin (Tf) and epidermal grown factor (EGF) receptors and the blood group (BG) antigen. We examined morphologically normal urothelium (34 cases), benign inflammatory and reactive conditions (24 cases), and atypical changes (20 cases) without exophytic or invasive neoplasia. The Ki67 nuclear antigen was used as the proliferation index and was complemented with the in vitro BrdU incorporation assay in 32 cases. The immunohistochemical reactions for Tf and EGF receptors and for the appropriate BG antigen were scored semi-quantitatively on frozen sections. We found that normal urothelium has very low Ki67 and BrdU indices as well as low reactivity for the two receptors and is almost invariably positive for the BG antigen. Benign conditions such as inflammation and metaplasia significantly augment the proliferation indices and the Tf receptor with little change in the EGF receptor and no significant effect on the BG antigen. Moderate atypia includes biologically heterogeneous cases which vary widely in proliferation and receptor expression. Severe atypia carcinoma in situ is consistently associated with markedly elevated proliferation indices, strong Tf receptor reactivity, and depressed BG antigen. The reactivity for EGF receptor is less consistently increased. Cases with a combination of strong EGF receptor reactions and absence of the expected BG antigen have a poor prognosis with progression to invasive cancer. PMID- 8229456 TI - Cytochrome P450 expression is a common molecular event in soft tissue sarcomas. AB - Two major xenobiotic metabolizing sub-families of cytochrome P450 (cytochrome P450 1A and cytochrome P450 3A) have been identified in soft tissue sarcomas. Cytochrome P450 1A was present in 70 per cent and cytochrome P450 3A was present in 78 per cent of tumours, respectively. A high proportion (86 per cent) of those tumours which contained cytochrome P450 1A or cytochrome P450 3A demonstrated co expression of both sub-families. In each tumour, cytochrome P450 immunoreactivity was identified in all tumour cells and there was no intra-tumour heterogeneity. These results indicate that expression of cytochrome P450 is a common molecular event in soft tissue sarcomas and that the presence of different sub-families of cytochrome P450 has implications both for the pathogenesis and for the treatment of these tumours. Cytochrome P450 expression may influence the intrinsic drug resistance of these tumours and also provide a molecular target for anti-cancer drugs which can be activated by cytochrome P450. PMID- 8229457 TI - Ultrastructural heterogeneity in undifferentiated bronchial carcinoma. AB - Electron microscopy is often suggested as a useful aid to the classification of light microscopically undifferentiated bronchial malignancies, features such as dense-core vesicles, desmosomes or tonofilaments, and microacini, allowing their designation as endocrine, squamous, or adenocarcinomas respectively. However, there is no reason to suppose that the heterogeneity of malignant bronchial tumours so often apparent by light microscopy or on immunolabelling might not occur at the ultrastructural level too. Extensive sampling of all deposits from eight subjects coming to necropsy with undifferentiated bronchial carcinoma revealed ultrastructural features of glandular and squamous differentiation to be widespread and often to occur together, although dense-core vesicles were not seen in any of the tumours studied. Heterogeneity was present within individual tumour deposits and particularly between different deposits of those tumours which had disseminated, such that any ultrastructural diagnosis would have been significantly influenced by sampling. Such variation should be borne in mind when ultrastructural features are used to classify bronchial malignancies. PMID- 8229458 TI - Three-dimensional reconstruction of human embryonic notochords: clue to the pathogenesis of chordoma. AB - Three-dimensional reconstruction experiments performed on serial sections of human embryos showed that the anatomy of the caudal and rostral ends of the notochord was complex. Forking of the ends, with separate fragments of chordal tissue, was demonstrated and these provide a way by which notochordal cell rests could be left behind in the basicranial and sacral regions when the notochord involutes elsewhere. Assuming the histogenesis of chordomas from notochordal cell rests, this would furnish an explanation for the observed skeletal distribution of chordomas. PMID- 8229459 TI - Clinical necropsy rates during the 1980s: the continued decline. AB - The trends in necropsy rates during the 1980s in three groups of British teaching hospitals in Leicester, Manchester, and Sheffield were compared in a retrospective study. The clinical necropsy rates declined in all three cities: in Leicester, from 16 to 10 per cent; in Manchester, from 14 to 8 per cent; and in Sheffield from 18 to 11 per cent. The medico-legal and overall necropsy rates showed variable trends between the cities. Specific events and changes in organization during the review period were correlated with the changing trends. The observed trends emphasize the continued decline in clinical necropsy rates over the last decade and illustrate the importance of monitoring differential necropsy rates. PMID- 8229460 TI - Electron microscopic study of the attachment and penetration of rabbit intestinal epithelium by Providencia alcalifaciens. AB - Ilea of adult rabbits with removable ileal ties (RITARD model) that developed diarrhoea and ileal loops of adult rabbits after inoculation with Providencia alcalifaciens, isolated from two patients with diarrhoea, were studied by transmission electron microscopy for enterocyte-bacterial interactions. Two modes of entry of bacteria into epithelial cells were seen: one directly by endocytosis associated with polymerization of cytoskeletal components and the other by disruption of tight junctions with entry into and proliferation in intercellular spaces. PMID- 8229461 TI - Apoptotic bodies in a murine model of alcoholic liver disease: reversibility of ethanol-induced changes. AB - The aim of this study was to investigate the effects of ethanol exposure on the location and number of apoptotic bodies in murine liver. Livers from 21 control mice were compared with those of 41 animals exposed to ethanol vapour by inhalation for up to 23 days. In the control livers, apoptotic bodies were identified in association with 36 per cent of the terminal hepatic venules examined and 88 per cent of these bodies were located in the first two rows of perivenular hepatocytes. By comparison, in the ethanol-exposed group, apoptotic bodies were observed in association with 72 per cent of the terminal hepatic venules examined and more were found further from them. This increase in the number of apoptotic bodies was shown to depend on the duration of ethanol exposure and, furthermore, the effects of ethanol were shown to be completely reversed by a period of abstinence. These results indicate that ethanol exposure has time-dependent, reversible effects on both the location and the number of apoptotic bodies in murine liver. This extends knowledge gained from previous work on rat livers. PMID- 8229462 TI - Lipofuscin is a photoinducible free radical generator. AB - Despite the association between high concentrations of the age pigment lipofuscin and various age-related pathologies, there is contention as to whether or not the intracellular accumulation of such granules has an adverse effect on cell function. In this paper, we present evidence to indicate that lipofuscin is a photoinducible free radical generator. White light irradiation of lipofuscin granules resulted in the production of superoxide anions, their rate of production increasing with increased light intensity. This effect appeared to be maximal in the blue light region of the spectrum. We speculate that lipofuscin could be detrimental to bodily functions through free radical generation initiated either by light or interaction with endogenous free radicals. PMID- 8229463 TI - Antioxidant activity of flavonoids: efficiency of singlet oxygen (1 delta g) quenching. AB - Flavonoids, polyphenolic pigments widely present in plants, have been reported to act as scavengers of various oxidizing species. However, most often an overall antioxidant effect was measured. In this paper we report the results of a systematic study of the reactivity of 13 selected flavonoids (from the flavonol, flavone, flavanone and flavane families) with singlet oxygen (1O2(1 delta g)) in order to establish a structure-activity relationship. The rate constants of the chemical reaction of these flavonoids with 1O2(k r) and their rate constants of 1O2 physical quenching (kq) have been determined by kinetic measurements and near IR singlet oxygen luminescence. The efficiency of the physical quenching is mainly controlled by the presence of a catechol moiety on ring B, whereas the structure of ring C (particularly the presence of a hydroxyl group activating the double bond) is the main factor determining the efficiency of the chemical reactivity of these compounds with 1O2. The total reactivity factor determining the efficiency of the chemical reactivity of these compounds with 1O2. The total reactivity scale is dominated by kq, which is in general higher than kr. (+) Catechin is the most efficient quencher of the series. PMID- 8229464 TI - Investigation of the usefulness of the photodynamic reaction for the purging of bone marrow from metastatic neuroblastoma cells. AB - The neuroblastoma cell lines SK-N-LO, SK-PN-DW and IMR 5 were stained with Merocyanine 540 and exposed to white light. 99% of clonogenic tumour cells were destroyed while 60% of bone marrow progenitor cells survived this treatment. PMID- 8229465 TI - Effects of low power 904 nm radiation on rat fibroblasts explanted and in vitro cultured. AB - In this study, 904 nm radiation was used on rat fibroblasts, explanted and in vitro cultured, to verify the action of various factors on cell growth. Parameters which can modify the behaviour of cultured cells, including the pulse repetition rate and the intensity of the radiation, the distance between the source and the irradiated dishes, the daily duration of exposure and the length of treatment, were studied. The most effective values of the intensity and pulse repetition rate which stimulate cell growth were 3 x 10(-4) W m-2 and 1.6 kHz respectively; the best stimulation was obtained at a sample-source distance of 0.1 m. The duration of daily exposure had no significant effect, whereas the best stimulus of cell growth was obtained by extending the treatment to 12 days. PMID- 8229466 TI - Lysosomes, a key target of hydrophobic photosensitizers proposed for photochemotherapeutic applications. AB - Despite their important biological activity, lysosomes have been generally neglected as important primary targets of photosensitizers, because they are not easily accessible for experiments. This paper reviews factors favoring the localization of photosensitizers in lysosomes and the various experimental approaches which have been used so far for the characterization of the lysosomal staining by various photosensitizing dyes, including porphyrins, chlorins and phenoxazines. The experimental difficulties observed in combining several in vitro techniques for the unambiguous demonstration of lysosomal targeting are examined. New data on tetraphenylporphine derivatives and a pyropheophorbide, as well as previous data on photofrin II, are presented to illustrate the advantages and possibilities of microspectrofluorometry in the study of photosensitizer localization in single living cells. Both spectral and topographic information available from areas smaller than 1 microns2 make it possible to characterize fairly specific sites of localization through the use of specific and vital fluorescent probes of lysosomes, such as Lucifer Yellow. It is also shown by microspectrofluorometry on single living cells that the chronology of the photosensitized reactions induced by specific or unspecific lysosomal photosensitizers can be easily followed. The photosensitized lipofuscin formation observed at the plasma membrane level with the lysosomotropic tetraphenylporphine supports the contention that it is very rare to find a truly specific lysosomal photosensitizer. PMID- 8229467 TI - Photobiological properties of hematoporphyrin diesters: evaluation for possible application in photochemotherapy of cancer. AB - The dimethyl, diethyl, dipropyl, dibutyl, diamyl, dihexyl and diheptyl esters of hematoporphyrin (Hp) were synthesized and shown to be more strongly retained on a reverse phase (C18) high performance liquid chromatography column than most components of Photofrin II (PII) - the sensitizer used for photochemical treatment of cancer in the clinic. The Hp diesters were found to be less efficient than PII in sensitizing cells to photoinactivation. This was partly due to de-esterification of the Hp diesters by esterase activity in the serum. The de esterification of the Hp diesters was highly dependent on the ester group, with Hp dimethyl ester (t1/2 for conversion to Hp monomethyl ester was 6 min) being de esterified with a rate 500 times faster than that for Hp diheptyl ester. Incubation of NHIK 3025 cells with these dyes showed that the Hp diesters were all partly located in extranuclear spots and partly diffusely distributed in the cytoplasm. The fluorescing spots may be due to lysosomally located Hp diesters, since the lysosomal marker enzyme beta-Nacetyl-D-glucosaminidase was partly inactivated by Hp diesters and light. PMID- 8229468 TI - Endogenous porphyrin distribution induced by 5-aminolaevulinic acid in the tissue layers of the gastrointestinal tract. AB - The accumulation of endogenous porphyrins in rats following systemic administration of 5-aminolaevulinic acid (ALA) has been examined to assess the photosensitization characteristics of this technique for photodynamic therapy (PDT) and chemical extraction assays with fluorescence and absorbance detection of the porphyrin content have been carried out. We compared the results obtained using quantitative microfluorimetry on normal gastric and colonic tissues in rats at 0.5, 1, 2, 4 and 6 h and chemically induced duodenal tumours 2 and 4.5 h after intravenous administration of ALA at a dose of 200 mg kg-1. With chemical extraction followed by high performance liquid chromatography analysis, protoporphyrin IX (PpIX) was found to be the predominant porphyrin present, reaching peak levels of several microgrammes per gramme at 2-4 h in each type of tissue; a small amount of coproporphyrin was detected at 0.5 and 2 h in normal gastric mucosa and duodenal tumour respectively. Both the extraction assay and quantitative microfluorimetry showed that the porphyrin fluorescence builds up rapidly in the mucosal layers of the colon and stomach, reaching a maximum at 2 h, whereas lower fluorescence levels were found with a slower rate of accumulation in the corresponding muscularis layers. A significant PpIX content was found in the duodenal tumour, with a maximum of 7.1 micrograms g-1 4.5 h after ALA administration. We conclude that systemic administration of ALA can induce effective tissue sensitization with protoporphyrin IX and appears to be a promising technique for PDT. PMID- 8229469 TI - Porphyrins, porphyrias, cancer and photodynamic therapy--a model for carcinogenesis. AB - Porphyrins are the only and most powerful photosensitizers synthesized internally. To understand better the involvement of porphyrins in photosensitization reactions, the heme biosynthetic pathway is first described, as well as the main features of its regulation in both erythroid and hepatic cells. Most disorders of porphyrin metabolism, known as porphyrias, are characterized by porphyrin accumulation. A full discussion of these diseases, their classification and relevant biochemical and clinical signs are presented. Abnormalities in heme biosynthesis in disorders other than porphyrias, such as iron-deficient and sideroblastic anemias, lead poisoning, hereditary tyrosinemia, chronic renal disease and alcoholism, are briefly considered. A complete survey of the experimental research on the biosynthesis of porphyrins in tumors and of the important association between cancer and porphyrias is dealt with. The link to photodynamic therapy (PDT) emerges naturally and this is treated from the point of view of using porphyrins endogenously formed by the tumors for their localization and PDT. Finally, considering the nature of the alterations occurring in heme metabolism in tumors, and porphyrias and their ubiquity, a model is discussed where the abnormality of heme synthesis is involved in the initiating lesion of carcinogenesis. The model strongly predicts that the incidence of cancer will be high in cells with abnormal heme metabolism, suggesting that porphyric patients may be at greater risk of the development of cancer. PMID- 8229470 TI - Murine pharmacokinetics and antitumor efficacy of the photodynamic sensitizer 2 [1-hexyloxyethyl]-2-devinyl pyropheophorbide-a. AB - The combination of the new photodynamic sensitizer 2-[1-hexyloxyethyl]-2-devinyl pyropheophorbide-a (HPPH) and laser light of wavelength 665 nm showed antitumor activity against two s.c.-implanted murine tumors. HPPH also sensitized normal mouse foot tissue to light but photosensitivity decreased rapidly with time after HPPH administration. Mechanistic studies revealed that HPPH induced little direct tumor cell toxicity but was an effective mediator of vascular photodamage. Pharmacokinetic studies following intravenous injection of 1 mg [14C]HPPH per kilogram revealed a biexponential decay with time, with plasma alpha and beta half-lives of 0.69 and 21 h respectively. Fecal excretion was the primary route of elimination. The highest levels of [14C]HPPH were found in the liver, which also showed the greatest long-term retention. The sequence of decreasing uptake levels was the liver, adrenals, lung, spleen, kidney, urinary bladder, heart, eye, skin, pancreas, muscle, testes, fat and brain. This distribution correlated with the relative blood perfusion rates in the tissues. PMID- 8229471 TI - Biodistribution of a methylene blue derivative in tumor and normal tissues of rats. AB - By using a chemical extraction procedure and confocal laser scanning fluorescence microscopy we have investigated the kinetic patterns of uptake and biolocalization of a methylene blue derivative (MBD) in tumors and various normal tissues of Wistar rats bearing fibrosarcoma (Leeds ovarian tumor) after intravenous injection of MBD (10 mg kg-1 body weight). Similar kinetics of accumulation and elimination of MBD fluorescence were found in tumor tissue and surrounding normal skin and muscle tissues. However, the tumor:skin and tumor:muscle ratios of the MBD fluorescence intensity were found to be 9 and 4, respectively, 4 h after intravenous injection, indicating selective uptake of MBD by the tumor tissue. MBD was localized on the walls of all the vessels and extensively in the area of neoplastic cellular and tumorigenic fibrous components in the tumor tissue. Interestingly, no MBD fluorescence could be detected in the metastatic neoplastic cells in the remote lymph nodes. In the skin, MBD was mainly distributed in the keratinized epithelium of the epidermis, hair follicles and their accessories, while little was found both in the epidermis and dermis. In most other tissues, the maximal fluorescence intensity of MBD was found 1-4 h after injection, after which it decreased dramatically to almost undetectable levels 120 h postinjection. Strong fluorescence of MBD was seen in the tracheal mucosal epithelium, while little fluorescence was noted in the transitional epithelium of bladder. The kinetics of biolocalization of MBD in some other tissues (liver, spleen, kidney, brain, muscle, lung, heart) were also studied. PMID- 8229472 TI - Prevalence of cerebral palsy among ten-year-old children in metropolitan Atlanta, 1985 through 1987. AB - The Metropolitan Atlanta Developmental Disabilities Study was a population-based study (1985 through 1987) to determine the prevalence of five developmental disabilities among 10-year-old children. The disabilities included cerebral palsy, mental retardation, visual impairment, hearing impairment, and epilepsy. The prevalence of cerebral palsy (CP) and a description of the children with CP are reported here. Using a record review approach, we identified 204 10-year-old children with CP (resulting in a prevalence of 2.3 per 1000). The rate of CP was significantly higher among boys (prevalence odds ratio = 1.5; 95% confidence interval = 1.1, 2.0), and the rate was also higher among black children than white children (prevalence odds ratio = 1.3; 95% confidence interval = 1.0, 1.7). Thirty-three of the children (16%) acquired CP postnatally; these children were more likely to be black or male. The gender and racial differences found for acquired CP were greater than those for congenital CP. Approximately 75% of the children had one of the other four disabilities studied; 65% of the children were mentally retarded, 46% had epilepsy, and 15% had a sensory impairment. Our multiple-source method of identifying children with CP gave us a population-based sample from which to determine the prevalence of the condition and to study factors that are associated with CP. PMID- 8229473 TI - Denys-Drash syndrome: relating a clinical disorder to genetic alterations in the tumor suppressor gene WT1. PMID- 8229474 TI - Treatment of streptococcal pharyngotonsillitis: reports of penicillin's demise are premature. PMID- 8229475 TI - Impaired ventilation in infants sleeping facedown: potential significance for sudden infant death syndrome. AB - Although accidental suffocation has been suggested as a cause of sudden infant death syndrome in infants who are found prone with their faces straight down, the occurrence and effects on breathing of this position in living infants are unknown. We studied prone sleeping infants to see whether they will sleep with their faces straight down; whether they can spontaneously change from this position; the way that thermal stimuli or bedding softness influences this change; and whether rebreathing or airway obstruction occurs. We studied 11 healthy infants, aged 0.2 to 6 months, while they slept on soft and on firm bedding. Most infants slept facedown spontaneously or when turned to this position. All could turn their heads readily but slept facedown for variable periods. All infants put their faces straight down more often after cold than after warm stimuli. Obstructive apnea occurred only once, but rebreathing occurred in all subjects. When the infants were in the facedown position, inspired carbon dioxide was three times greater on soft than on hard bedding. End tidal carbon dioxide partial pressure rose in all subjects while they were in the facedown position; in one infant, high end-tidal carbon dioxide partial pressure and desaturation occurred without signs of arousal. We conclude that infants sleeping facedown may occasionally have significant asphyxia; this sleep position may have a role in some infant deaths diagnosed as sudden infant death syndrome. PMID- 8229476 TI - Home event recordings of oxygenation, breathing movements, and heart rate and rhythm in infants with recurrent life-threatening events. AB - For an evaluation of the usefulness of event recording in identifying mechanisms for apparent life-threatening events, 94 infants (median age, 3.1 months; range, 0.5 to 12 months) with a history of two or more apparent life-threatening events of hitherto unknown cause underwent event recording of transcutaneous oxygen pressure, arterial oxygen saturation, (not recorded in all patients), breathing movements, pulse waveforms, electrocardiogram, and instantaneous heart rate. Recordings were triggered by a fall in transcutaneous oxygen pressure to less than 20 mm Hg. During a median duration of event recording of 1.3 months (0.1 to 10 months), 52 events were recorded in 34 patients; 7 of the events had to be excluded because of uninterpretable signals. The following mechanisms were identified in the remaining 30 patients (45 events): a sudden change in skin perfusion but without hypoxemia (6 in 5 patients), hypoxemia induced by an epileptic seizure (6 in 5 patients), hypoxemia induced by suffocation by a parent (4 in 4 patients), and parental fabrication of events and medical history (7 in 6 patients). In the remaining 22 events in 12 patients, the precise mechanism of events could not be identified. Analysis of these as-yet-unexplained events showed prolonged abnormal hypoxemia (a fall in transcutaneous oxygen pressure to between 4 and 18 mm Hg and a fall in arterial oxygen saturation to 5% to 75%), lasting for 40 to 500 seconds in all. Only five of these events involved prolonged (> 20 seconds) apneic pauses, and only four an episode of bradycardia (heart rate < 80 or 60 beats/min). Thus event recording identified various mechanisms of apparent life-threatening events. In the events that remained unexplained, prolonged apneic pauses or episodes of bradycardia were found in only a minority. The identification of hypoxemia therefore may be more relevant to the early detection of these events than the identification of apnea or bradycardia or both. The relevance of these findings with regard to sudden infant death syndrome remains to be determined. PMID- 8229477 TI - Relation of infant feeding practices, cigarette smoke exposure, and group child care to the onset and duration of otitis media with effusion in the first two years of life. AB - The relation of infant feeding practices, cigarette smoke exposure, and group child care to the onset and duration of otitis media with effusion (OME) was evaluated in a cohort of 698 healthy infants prospectively monitored by tympanometry in the home every 2 to 4 weeks until 2 years of age. Except for an experimental group of children who were offered early tube placement, the study children received conventional care from their personal physician or clinic. We used LISREL, a structural equation modeling procedure (computer software), to explore associations between environmental variables and OME onset and duration while controlling for interrelations among the variables. Supine feeding position and early initiation of group child care were associated with earlier onset of OME. Shorter duration of breast-feeding, increased packs of cigarettes smoked per day in the home, and increased hours per week in group child care were associated with an increase in the amount of time with OME during one or more of the age blocks studied (birth to 6, 6 to 12, 12 to 18, and 12 to 24 months). For a decrease in the amount of time with OME during the first 2 years of life, prolonged breast-feeding and upright feeding position should be encouraged, and cigarette smoke exposure should be minimized. Limiting early child care in large groups might also be advisable. PMID- 8229478 TI - Neuropsychologic effects of stroke in children with sickle cell anemia. AB - We examined the nature and extent of neuropsychologic impairment in children with sickle cell anemia (SCA) with or without stroke. Twenty-nine children with SCA received cranial magnetic resonance imaging. Strokes were classified into three groups: diffuse cortical stroke (n = 11), anterior stroke (n = 6), or none (n = 12). Children with SCA and 20 age-matched sibling control subjects then received a neurologic examination and a neuropsychologic battery of tests that included motor, verbal, spatial, attentional, and memory measures. Tests of spatial function showed that children with diffuse cortical strokes were impaired, whereas children with anterior lesions had intrusions of irrelevant material during list recall. There were no significant differences between children with stroke and sibling control subjects on motor, verbal, or memory measures. Six children had evidence of stroke on magnetic resonance imaging without any history of a damaging neurologic event. These children had impaired neuropsychologic performance relative to that of sibling control subjects in a pattern similar to that of children with overt stroke. Children with SCA without stroke did not differ from sibling control subjects on any measure. Our results indicate that overt and silent strokes result in lesion-specific neuropsychologic deficits in children with SCA. PMID- 8229479 TI - Transient ischemic cerebral lesions during induction chemotherapy for acute lymphoblastic leukemia. AB - Ninety children were treated for acute lymphoblastic leukemia or non-Hodgkin lymphoma during 1986 through 1992 in the Children's Hospital, University of Helsinki, in Finland. During induction chemotherapy, nine of the children had visual hallucinations progressing to confusion and seizure. The symptoms were often preceded by severe constipation and significantly elevated blood pressure. Neuroradiologic examinations showed bilateral cortical or subcortical white matter lesions. Despite the stroke like manifestations, the lesions were reversible. The triangular shape and location of the lesions in the watershed areas between the major cerebral arteries suggest vascular ischemia as the cause. PMID- 8229480 TI - Outbreaks of astrovirus gastroenteritis in day care centers. AB - OBJECTIVE: This study evaluated astrovirus as a cause of diarrhea outbreaks among infants and toddlers in day care centers. DESIGN: Stool specimens were collected weekly during four periods (from January 1986 through December 1991) from children 6 to 30 months of age who were enrolled in prospective studies of diarrhea in day care centers. All diarrheal stool specimens were tested for bacterial enteropathogens, rotavirus, enteric adenovirus, and Giardia lamblia. A total of 1365 stool specimens from 70 outbreaks in which no etiologic agent was identified and from another 11 outbreaks with a known cause were tested for astrovirus, by means of a monoclonal antibody-based enzyme immunoassay. Confirmatory testing was performed by reverse transcriptase-polymerase chain reaction with primers designed to produce an 89 base-pair product. RESULTS: Astrovirus was detected in 6 (7%) of the 81 outbreaks. Of 217 children tested, 73 (34%) were infected with astrovirus; infections in 35 (48%) were symptomatic and in 38 (52%) asymptomatic. The six outbreaks lasted 11 to 44 days (median 22 days). Astrovirus excretion was detected for a duration of 2 to 30 days, with excretion occurring from 1 to 8 days (median 2 days) before diarrhea began to 1 to 20 days (median 2 days) after diarrhea ceased. Younger children (< or = 12 months) were at greater risk than older children (p = 0.011) of becoming infected with astrovirus during an outbreak and were more likely (p = 0.015) to have symptoms when infected. Of 24 specimens with astrovirus by enzyme immunoassay, 20 (83%) were confirmed to have the virus by reverse transcriptase-polymerase chain reaction. CONCLUSION: Astrovirus was an important cause of outbreaks of diarrhea among children attending day care centers, more frequently infected younger children, and often produced asymptomatic infections. PMID- 8229481 TI - First-phase insulin release in normal children. AB - Normal values for the first-phase insulin release during an intravenous glucose tolerance test are not yet well defined for children and adolescents. In this study, 69 normal subjects (aged 7 to 22 years) who had no family history of type I diabetes, a normal glycohemoglobin value, and a negative islet cell antibody test result underwent a standard intravenous glucose tolerance test. The mean (+/ SEM) first-phase insulin release increased with age and pubertal status: 7 to 10 years, 93 +/- 10.1 mIU/L; 11 to 15 years, 172.7 +/- 22.3 mIU/L; and 16 to 22 years, 163 +/- 28.5 mIU/L. The mean intraindividual variability in 11 subjects who underwent a second test was 23.6%. Acute stress, as estimated by observer assessment or by blood catecholamine levels, did not significantly correlate with first-phase insulin release. We conclude that first-phase insulin release is markedly lower in prepubertal children than in adolescents and young adults. PMID- 8229482 TI - Immune globulin administration as an approach to prevention of acute otitis media. PMID- 8229483 TI - Growth hormone for children with Down syndrome. PMID- 8229484 TI - Determination of the urinary D/L trans-3-hydroxyproline ratio: a noninvasive screening test for Alport syndrome. AB - Because disturbed conformation of connective tissue proteins can be accompanied by increased racemization (i.e., an increased ratio of dextrorotatory (D) to levorotatory (L) amino acid molecules), we studied by high-performance liquid chromatography the renal excretion of the D-form of the basement membrane specific trans-3-hydroxyproline in patients with Alport syndrome. The D/L ratio was significantly higher in patients with Alport syndrome than in patients with other renal diseases or in healthy control subjects. We therefore suggest that this factor may be a simple noninvasive (screening) test for Alport syndrome. PMID- 8229485 TI - Chronic intestinal pseudo-obstruction with meconium ileus at onset. AB - Cystic fibrosis is most often the underlying cause of meconium ileus. We describe the diagnosis and treatment of a patient with chronic intestinal pseudo obstruction, and not with cystic fibrosis, whose initial manifestation was meconium ileus. PMID- 8229486 TI - Atrophie blanche with onset in childhood. AB - Atrophie blanche (livedo vasculitis) is a superficial thrombotic condition characterized by grouped and reticulated erythematous and purpuric macules, painful ulcers, and atrophic scars, and is usually found in middle-aged women. We describe an 8-year-old boy with atrophie blanche. Therapy with antiplatelet medications seemed to alleviate pain and decrease the ulceration. PMID- 8229487 TI - A multicenter randomized trial comparing two surfactants for the treatment of neonatal respiratory distress syndrome. National Institute of Child Health and Human Development Neonatal Research Network. AB - OBJECTIVE: To compare the efficacy of two surfactants, Exosurf Neonatal (Burroughs Wellcome Co.) and Survanta (Ross Laboratories), for the treatment of neonatal respiratory distress syndrome. DESIGN: Multicenter randomized trial. SETTING: Eleven tertiary care university neonatal intensive care units participating in the National Institute of Child Health and Human Development Neonatal Research Network. PATIENTS: Newborn infants (n = 617) weighing 501 to 1500 gm with respiratory distress syndrome who were receiving assisted ventilation with 30% oxygen or more within 6 hours of birth were enrolled between January 1991 and January 1992. INTERVENTIONS: Infants were randomly assigned to receive up to four intratracheal doses of either Exosurf Neonatal (n = 309) or Survanta (n = 308). MAIN OUTCOME MEASURES: The occurrence of death or bronchopulmonary dysplasia 28 days after birth and the average fraction of inspired oxygen (FIO2) and mean airway pressure (MAP) during the first 72 hours after treatment. RESULTS: Death or bronchopulmonary dysplasia occurred in 67% of the infants in the Exosurf group and 62% of those in the Survanta group (adjusted relative risk, 1.07; 95% confidence interval, 0.96 to 1.20). During the 72 hours after the first surfactant dose, the average FIO2 (+/- SEM) was 0.50 +/- 0.01 for Exosurf and 0.42 +/- 0.01 for Survanta (difference, 0.08; 95% confidence interval, 0.05 to 0.11); the average MAP (+/- SEM) was 7.64 +/- 0.21 cm H2O for Exosurf and 6.93 +/- 0.21 cm H2O for Survanta (difference, 0.71 cm H2O; 95% confidence interval, 0.13 to 1.29 cm H2O). There was no difference between the groups in the incidence of other neonatal morbidities or in the duration of hospitalization, assisted ventilation, or supplemental oxygen administration. CONCLUSION: We found no difference between treatment groups in the incidence of death or bronchopulmonary dysplasia, although we did observe a difference in the initial response to treatment as measured by FIO2 and MAP. PMID- 8229488 TI - Arterial oxygenation and pulmonary arterial pressure in healthy neonates and infants at high altitude. AB - We sought to document arterial oxygen saturation relative to changes in the right ventricular pressure/left ventricular pressure ratio (RVP/LVP ratio), an index of pulmonary arterial pressure, in infants born at high altitude. We performed pulse oximetry and echocardiography in 15 healthy infants born in Leadville, Colo. (3100 m), at 6 to 24 hours, 24 to 48 hours, 1 week, 2 months, and 4 months of age. Pulse oximetry was done under conditions of wakefulness, feeding, and active and quiet sleep. All infants received supplemental O2 at delivery and during postnatal transition; all oximetry measurements were performed with infants breathing room air. The mean arterial O2 saturation ranged from 80.6% +/- 5.3% to 91.1% +/- 1.7% during the 4 months. Values fell during the first week after birth and then rose gradually to attain near-birth values at 2 and 4 months of age. Arterial O2 saturation was uniform among behavioral states at 6 to 24 hours and 24 to 48 hours of age. After 1 week of age, values were highest during wakefulness, intermediate during feeding and active sleep, and lowest during quiet sleep. The RVP/LVP ratio remained in the normal or mildly elevated range throughout the study period. We conclude that the RVP/LVP ratio promptly becomes normal at high altitude, and despite low arterial O2 saturation in the first weeks to months after birth, healthy newborn infants at 3100 m show little evidence of acute pulmonary hypertension. PMID- 8229489 TI - Breast-feeding of very low birth weight infants. AB - Twenty very low birth weight infants (birth weight < 1500 gm) were assessed to compare the clinical effects of breast and bottle feedings. The infants started breast-feeding during the same week that they started bottle feedings. Five breast-feedings and five bottle feedings for each infant were observed. Axillary temperature and weight before and after feedings were measured, and oxygen saturation, respiratory rate, and heart rate were monitored and recorded every 2 minutes during the feeding periods. The results showed no difference in oxygen saturation during breast-feeding (p = 0.056) but a lower incidence of oxygen desaturation (< 90%) (21% vs 38% in breast-feeding vs bottle feeding, respectively; p < 0.025). Infants with bronchopulmonary dysplasia had higher oxygen saturation during breast-feeding than during bottle feeding (p < 0.025), but weight gain during breast-feeding sessions was less (median, no gain vs 31 gm, p < 0.001). We conclude that (1) very low birth weight infants can tolerate both breast and bottle feedings at the same postnatal age; (2) very low birth weight infants are less likely to have oxygen desaturation to less than 90% during breast-feeding; and (3) weight gain is less during breast-feeding, probably because of lower intake, and may require more lactation counseling or supplementation of the feedings. PMID- 8229490 TI - Congenital cytomegalovirus infection and neonatal auditory screening. AB - Auditory screening of newborn infants has been recommended on the basis of the presence of risk criteria, including congenital infection. We assessed the ability of risk criteria-based neonatal auditory brain stem response to identify infants with hearing loss resulting from congenital cytomegalovirus (CMV) infection. Data from 6 1/2 years of risk criteria-based neonatal auditory screening were compared with the results of screening of all newborn infants for congenital CMV infection. Infants with congenital CMV infection received follow up hearing evaluations. Congenital CMV infection was found in 167 (1.3%) of 12,371 infants; 134 had follow-up hearing evaluations, and 14 (10.4%) had confirmed sensorineural hearing loss. The rate of sensorineural hearing loss resulting from congenital CMV infection was 14 per 12,371 infants, of 1.1 per 1000 live births; the rate of bilateral loss > or = 50 dB was 0.6 per 1000. Although 2036 infants received auditory screening because of risk criteria, only 34 (20%) of 167 infants with congenital CMV infection were included. Only 2 (14%) of 14 children with sensorineural hearing loss caused by CMV were identified by risk criteria-based screening. We conclude that congenital CMV infection is an important cause of hearing impairment. Neonatal auditory screening based on the presence of risk criteria will fail to identify the majority of cases of sensorineural hearing loss caused by congenital CMV infection. PMID- 8229491 TI - Comparison of two preservation solutions for erythrocyte transfusions in newborn infants. AB - To determine whether one of the newer preservation solutions for packed red blood cells (PRBC) is safe and effective in the transfusion of the very low birth weight infant, we conducted a randomized trial comparing PRBC preserved with the anticoagulant solution mannitol-adenine-dextrose (AS-1) and PRBC preserved with citrate-phosphate-dextrose-adenine (CPDA-1). Sixteen infants (birth weight 863 +/ 218 gm) with a gestational age of 26 +/- 3 weeks received one to three small volume replacement transfusions with PRBC, 17 ml/kg, preserved with either AS-1 or CPDA-1 in a double crossover design. Transfusion with AS-1-preserved PRBC resulted in an equivalent increase in hemoglobin concentration when adjustment was made for the difference in the hemoglobin concentration of the transfused PRBC. During the transfusion, the percentage decrease in serum glucose values was greater with the CPDA-1 preservative than with the AS-1 preservative (54% +/- 13% vs 42% +/- 11% at 1 hour; p < 0.001). No other significant difference in blood chemistry values was found. Urine output was unaffected by AS-1 in the posttransfusion period. We conclude that (1) small-volume PRBC transfusions with AS-1 can be used in the very low birth weight infant without apparent detriment, (2) AS-1-preserved cells are as effective as cells preserved with CPDA-1 for increasing hemoglobin concentration, and (3) the higher dextrose content of the AS-1-preserved blood allows for improved glucose homeostasis during transfusion. PMID- 8229492 TI - Ultrasound guidance of suprapubic bladder aspiration in neonates. AB - We performed a prospective, randomized trial comparing ultrasound guidance of suprapubic bladder aspiration with unguided aspiration. The acquisition of urine improved with ultrasound guidance from 60% to 96.4%. The number of needle insertions decreased and the amount of urine obtained increased. Ultrasonography is a useful tool for bladder aspiration. PMID- 8229493 TI - Response to Haemophilus influenzae type b conjugate vaccine in chronically ill premature infants. AB - Twenty-two premature infants with chronic lung disease (median gestational age 28 weeks) received polyribosylribitol phosphate-outer membrane protein conjugate Haemophilus vaccine at 2 and 4 months of chronologic age. The proportions with antibodies to polyribosylribitol phosphate at levels > 1 microgram/ml after doses 1 and 2 were 27% and 55%; geometric mean titers were 0.43 and 0.73 microgram/ml, significantly lower than values for term infants. PMID- 8229494 TI - Comparison of arterial and capillary bilirubin values in neonates with arterial lines. AB - To determine whether arterial and capillary values for bilirubin agree in neonates, we analyzed 35 simultaneously sampled capillary and arterial specimens from 13 infants. We found a high correlation between arterial and capillary bilirubin values (r = 0.993). We conclude that arterial bilirubin samples may be substituted for capillary samples in newborn infants. PMID- 8229495 TI - Plasma ammonia levels in very low birth weight preterm infants. AB - The mean plasma ammonia level at birth of 36 very low birth weight infants (< or = 32 weeks of gestation) was 71 +/- 26 mumol/L (121 +/- 45 micrograms/dl), which is similar to the mean level in preterm infants born at > or = 32 weeks of gestational age. Plasma ammonia levels declined to 42 +/- 14 mumol/L (72 +/- 24 micrograms/dl) at 7 days of age; mean ammonia levels at 14, 21, and 28 days of age were similar to that at 7 days of age and to the mean plasma ammonia level of 14 healthy term infants at birth (45 +/- 9 mumol/L (77 +/- 16 micrograms/dl)). PMID- 8229496 TI - Bacterial polysaccharide immune globulin for prophylaxis of acute otitis media in high-risk children. AB - We evaluated the prevention of recurrences of acute otitis media (AOM) by bacterial polysaccharide immune globulin (BPIG), a hyperimmune human immune globulin prepared by immunizing donors with bacterial polysaccharide vaccines. We used a randomized, stratified, double-blind, placebo-controlled design. Children < or = 24 months of age with 1 to 3 prior episodes of AOM received BPIG, 0.5 ml/kg, or saline placebo intramuscularly at entry and 30 days later. During the 120-day follow-up period, AOM was diagnosed by using clinical criteria and was confirmed with tympanocentesis and culture of the middle ear exudates. Eighty eight episodes of AOM were observed in 76 patients who completed the study. The incidence of AOM during the entire 120-day study period was similar in BPIG and placebo recipients. Pneumococcal AOM was significantly less frequent in BPIG recipients (0.21 episode per patient) than in placebo recipients (0.45 episode per patient; p = 0.05). Time spent free of AOM was significantly prolonged in recipients of BPIG, in comparison with placebo recipients (51 vs 35 days; p = 0.034). This study demonstrated that circulating antibody, even without stimulation of specific local immunity, may prevent infection of the middle ear. The use of immune globulin preparations for longer periods or at a higher dosage might decrease the incidence of recurrent AOM in otitis-prone children, and deserves further evaluation. PMID- 8229497 TI - Chlorpromazine with and without lorazepam as antiemetic therapy in children receiving uniform chemotherapy. AB - We prospectively studied the efficacy and adverse effects of chlorpromazine (30 mg/m2 given intravenously) plus lorazepam (0.04 mg/kg given intravenously) versus chlorpromazine alone in a controlled, double-blind, randomized, parallel-design investigation in 25 children (1.5 to 17.3 years of age) with acute lymphoblastic leukemia. Response to other antiemetics in eight children refusing random assignment to treatment was also evaluated. All children were receiving intravenous infusions of teniposide plus cytarabine, the pharmacokinetics of which were characterized for each of the one to four courses. There were no differences between the 11 patients randomly assigned to receive chlorpromazine alone and the 14 randomly assigned to receive lorazepam plus chlorpromazine in the number of emesis episodes (6.0 vs 5.9; p = 0.53), frequency of dystonic reactions (3% vs 5%), or akathisia (13 vs 10%). The only serious adverse event, symptomatic hypotension, occurred in a boy receiving chlorpromazine plus lorazepam. An exploratory pharmacodynamic analysis revealed that the only variable that correlated with vomiting was cytarabine 1 1/2-hour plasma concentration (p = 0.007). Children who received either chlorpromazine plus lorazepam or chlorpromazine alone had fewer episodes of vomiting than those who received "conventional" antiemetic therapy (6.0 vs 8.6; p = 0.01). We conclude that the severity of emesis is related to the plasma concentration of cytarabine; that intravenously administered chlorpromazine is as effective as chlorpromazine plus lorazepam in preventing chemotherapy-induced vomiting; and that the potential for adverse effects with the addition of lorazepam may be a disadvantage. PMID- 8229498 TI - A randomized, double-blind study comparing cefixime and trimethoprim sulfamethoxazole in the treatment of childhood shigellosis. AB - We compared the clinical and bacteriologic response of 5-day treatment with cefixime, 8 mg/kg per day, with the response to trimethoprim-sulfamethoxazole (TMP-SMX), 10-50 mg/kg per day, the currently recommended therapy. Of the assessable children with acute, culture-proven shigellosis, 38 received cefixime and 39 received TMP-SMX. Pretreatment data on the two study groups were similar. In the first group, all isolates were susceptible to cefixime; in the TMP-SMX group, 32 isolates were resistant and 7 were susceptible to TMP-SMX. Clinical response (day 5) showed cure, improvement, and failure in 89%, 8%, and 3%, respectively, of the cefixime group, and in 25%, 44%, and 31%, respectively, of the TMP-SMX-resistant group (p < 0.001). Bacteriologic cure (day 3) occurred in 78% and 23% of the cefixime and TMP-SMX-resistant groups, respectively (p < 0.001). Clinical or bacteriologic relapse (day 12) was infrequent in both groups. The response to treatment of the cefixime and the TMP-SMX-susceptible groups was similar. No significant side effects were noted. We conclude that cefixime is superior to TMP-SMX in the treatment of suspected shigellosis in areas with a high rate of resistance to TMP-SMX. PMID- 8229499 TI - Comparative efficacy of two- and five-day courses of ceftriaxone for treatment of severe shigellosis in children. AB - Administration of ceftriaxone for 5 days has been shown to be highly efficient in the treatment of severe shigellosis in children. Our study, involving 40 children, demonstrated that a 2-day course of ceftriaxone was as beneficial as a 5-day course and constituted effective therapy for children with severe shigellosis. PMID- 8229500 TI - Copper-histidine therapy for Menkes disease. AB - Menkes disease is an X-linked genetic disorder of copper transport that results in death from severe progressive neurodegeneration by the age of 3 years. We report here our 17 years' experience with the treatment of Menkes disease with subcutaneous administration of copper-histidine. Two patients (16 and 6 years of age) whose therapy was begun within 1 month of birth have done well neurologically. The other five patients have done poorly despite treatment initiated at 2 to 7 months of age. Copper-histidine therapy may be an effective treatment if started early. PMID- 8229501 TI - Daniel C. Darrow. PMID- 8229502 TI - Effect of filtration on fluoride concentrations in water. PMID- 8229503 TI - Neonatal sepsis caused by a new group B streptococcal serotype (type V) PMID- 8229504 TI - Interleukin-8 in pulmonary effluent fluid of preterm infants. PMID- 8229505 TI - Behavioral manifestations of hypothyroidism versus thyroxine effects. PMID- 8229506 TI - Efficacy of cough suppressants. PMID- 8229507 TI - Alpha interferon therapy in children with chronic active hepatitis B and delta virus infection. AB - Two children with chronic hepatitis B virus and delta virus infection were treated with alpha interferon. Both tolerated the therapy without complications, and one demonstrated clearance of the infection and development of immunity. PMID- 8229508 TI - Galactosemia: the uridine diphosphate galactose deficiency-uridine treatment controversy. PMID- 8229509 TI - Measurements of uridine diphosphate hexoses in galactosemia. PMID- 8229510 TI - Changes of serum lactate concentration, cardiac output, and heart rate induced by blood transfusions in preterm infants. PMID- 8229511 TI - Aminotransferase activity in celiac disease. PMID- 8229512 TI - Aminotransferase activity in celiac disease. PMID- 8229513 TI - Insulin dependency after transient neonatal diabetes. PMID- 8229514 TI - Prognostic factors in childhood epilepsy. PMID- 8229515 TI - The pineal gland in human beings: relevance to pediatrics. AB - Several facts suggest that the pineal gland must have a significant role in human beings: the presence of melatonin secretion from infancy to old age; a circadian secretory pattern similar to that found in animal species in which this gland has well-defined functions; its responsiveness to light; and the presence of melatonin receptors in the hypothalamus. Despite the importance of the pineal gland and melatonin in the reproductive activity of all nonprimate vertebrate species studied, the relationship of melatonin secretion to the activity of the hypothalamic-pituitary-gonadal axis in human beings remains presumptive. Some data suggest a possible pineal-reproductive connection throughout the human lifespan, but new research approaches are needed for a better definition of this connection during normal development and in disorders of the reproductive axis. In addition, recent studies that have applied strictly defined techniques to unravel the masking effects of various behavioral and environmental factors suggest that the pineal gland and melatonin have a fundamental role in the regulation of the human biologic clock. Melatonin concentrations in blood or urine may become a useful marker of the circadian rhythm in disorders of rhythms. Moreover, administration of melatonin in physiologic or pharmacologic doses may have an important application in suppression of the hypothalamic-pituitary gonadal axis as a contraceptive, and in therapy for disorders of biologic rhythms. Among the latter, of particular interest to the pediatric population will be the potential application of melatonin treatment in establishing or reestablishing circadian rhythms in infants and children maintained for long periods under artificial light conditions, as encountered in intensive care units, and in the treatment of sleep and other rhythm disorders associated with developmental delay or blindness. Further research and approval by the U.S. Food and Drug Administration will be required before these applications will be available to the practicing pediatrician. The identification of melatonin receptors in the human hypothalamic fetal and adult tissues and the development of potent melatonin agonists and antagonists have paved the way for new research approaches that are needed to further investigate this intriguing, long-forgotten gland in human beings. PMID- 8229516 TI - A policy regarding research in healthy children. PMID- 8229517 TI - School lunch: a comparison of the fat and cholesterol content with dietary guidelines. AB - OBJECTIVE: To compare the fat and cholesterol content of the foods offered and selected in an elementary school lunch program with current dietary guidelines. DESIGN: For 105 school days we recorded the food items selected by elementary school students in an entire school district (262,851 meals) who were given a choice between two entrees. The nutrient content of foods was assessed with a computerized nutrient data base supplemented by the food manufacturers' data. SETTING: Sixteen elementary schools in the Bellevue (Washington) School District. PARTICIPANTS: The number of students eating school lunch averaged 2500 per day, of whom 25% were from households with incomes less than 185% of poverty. INTERVENTION: None. MAIN OUTCOME MEASURES: We determined the nutritional content of the average meal selected; the proportion of days when one of the two offered entrees met fat and cholesterol guidelines; and the proportion of children selecting the entrees that met the guidelines. RESULTS: The average lunch selected had 35.9% of calories from total fat and 12.6% from saturated fat, exceeding the guidelines of 30% and 10%, respectively. Lunch contained an average of 57 mg cholesterol (106 mg/1000 kcal) and met guidelines. One of the two daily entree choices met guidelines for both total fat and saturated fat on 20% of days, and met both fat and cholesterol guidelines on 14% of days. When available, entrees meeting the fat guidelines were chosen by 37% of students, and entrees meeting both fat and cholesterol guidelines were chosen by 34% of students. CONCLUSIONS: In this school district the average lunch selected did not meet the current guidelines for dietary fat; when given the choice, more than one third of students selected the entrees that met these guidelines. PMID- 8229518 TI - Predictors of axial and peripheral bone mineral density in healthy children and adolescents, with special attention to the role of puberty. AB - The role of various physical and lifestyle factors in determining axial bone mineral density (BMD) at the lumbar segment of the spine, as measured by dual photon absorptiometry, and peripheral BMD at the distal third of the radius, as measured by single-photon absorptiometry, was assessed in 299 healthy white children of both sexes, aged 6 to 18 years. The BMD measurements were correlated with age, height, weight, body mass index, and pubertal status. Peripheral and axial BMD were highly correlated with age, height, weight, and pubertal stage, and more weakly with body mass index. Approximately 76% of the observed changes in peripheral BMD were accounted for by age, height, weight, and pubertal stage, with weight being the single strongest predictor. Up to 80% of the variation in axial BMD was explained by weight and pubertal stage, with pubertal stage being the strongest single predictor. After adjustment for weight, the effect of puberty on axial BMD in both sexes was greatest between middle and late puberty. These data indicate that a large amount of the observed changes on BMD is accounted for by standard measures of growth and development, which are largely genetically determined. Peripheral BMD rose steadily with age. Axial BMD increased steadily before puberty, followed by accelerated increases during puberty, beginning at 10 years of age in girls and 13 years of age in boys. A significant positive effect of dietary calcium intake on peripheral BMD and of physical activity on axial BMD indicated a potentially important impact of physical activity and calcium intake on peak bone mass. PMID- 8229519 TI - Blood pressure nomograms for children and adolescents, by height, sex, and age, in the United States. AB - Because height is a more appropriate index of maturation than weight for use with normative blood pressure (BP) data, we developed normative BP levels for children, by sex, while accounting for age and height simultaneously. Eight U.S. studies used in the Report of the Second Task Force on Blood Pressure Control in Children and one additional study of BP in U.S. children were reanalyzed to develop age-sex-height-specific values for normative BP values among 56,108 children, aged 1 to 17 years, seen at 76,018 visits. Height percentiles were computed on the basis of standard National Center for Health Statistics growth charts. When height is taken into account, more short children (10th age-sex specific height percentile) and fewer tall children (90th age-sex-specific height percentile) are likely to be classified as hypertensive than when the current age sex-specific percentiles of BP alone are used. Tables are provided for boys and girls separately, by single year of age (1 to 17 years) and by the 90th and 95th percentiles of systolic blood pressure and diastolic blood pressure (fifth phase of Korotkoff sounds) for selected age-sex-specific height percentiles based on standard U.S. growth charts. PMID- 8229520 TI - Decision rules for predicting vaccination status of preschool-age emergency department patients. AB - We produced and tested rules to predict undervaccination among preschool-age emergency department (ED) patients. Data were gathered on demographics, vaccination status, health status, and health care utilization from parents, ED physicians, and ED charts at an urban teaching hospital in Rochester, N.Y. Primary care charts were reviewed to verify vaccination status. Using recursive partitioning, we developed decision rules to predict undervaccination. Decision rules were developed on a sample of 602 ED patients 4 to 48 months of age and then prospectively tested on 1832 ED patients aged 6 to 36 months. Factors associated with undervaccination for any vaccine included parental report of vaccination delay (odds ratio = 8.1; p < 0.001), inability to report the receipt of the appropriate number of vaccines (odds ratio = 4.5; p < 0.001), lack of health insurance (odds ratio = 3.6, p < 0.001), elapsed time since the last visit to primary care provider (p < 0.001), household size (p < 0.001), and maternal age (p < 0.01). Eight decision rules were produced that varied in their number of questions (one to six), sensitivity (0.27 to 0.87), and specificity (0.54 to 0.98). No single rule was both highly sensitive and highly specific. The rules' sensitivities and specificities were similar for the validation sample of 1832 patients. Thus a decision rule could not be produced that was both sensitive and specific. Identification of undervaccinated children by means of information available at an ED visit is inherently difficult. Interventions in the ED may be inefficient unless better methods of assessing vaccination status can be developed. PMID- 8229521 TI - Prevalence of von Willebrand disease in children: a multiethnic study. AB - OBJECTIVE: Von Willebrand disease (vWD) was thought to be a rare disorder until a recent survey reported a prevalence of 0.8% in an ethnically homogenous community in northern Italy. The purpose of this study was to determine the prevalence of vWD in an ethnically heterogenous population. METHODS: Von Willebrand factor (vWF) was measured by the ristocetin cofactor method in 600 healthy children, aged 2 to 18 years, seen for routine school physical examinations in a three state region. Personal and family bleeding symptoms were determined by questionnaire. The diagnosis of vWD required a personal history of bleeding symptoms, an abnormal vWF activity concentration, and a family history of at least one immediate family member with bleeding symptoms. RESULTS: A total of 315 subjects were white, 212 were black, 16 were Hispanic, 10 were from other groups, and 47 were biracial. Eight subjects (four black, four white) met the criteria for vWD, for a prevalence of 1.3%. Seven subjects with vWD had blood group O (mean vWF = 32 U/dl; range, 10 to 42 U/dl), and one had blood group A (vWF = 41 U/dl). Children who had blood group O had significantly (p < 0.001) lower vWF activities (median, 83 U/dl, range, 43 to 162 U/dl) than those from non-O blood groups (median, 98 U/dl; range, 51 to 190 U/dl). There were no significant differences in vWF activity by ethnicity. The vWF activity was significantly (p < 0.02) greater for boys than girls in both blood groups. CONCLUSION: Von Willebrand disease is the most common congenital hemostatic disorder; its high prevalence is not limited to one ethnic group. PMID- 8229522 TI - Age-related changes in serum free thyroxine during childhood and adolescence. AB - The purposes of this study were to reevaluate age-related changes in the concentration of serum free thyroxine (T4) between early infancy and adulthood, and to study the relationship of serum thyrotropin to these changes in free T4. Five hundred thirty-six healthy infants, children, adolescents, and adults between 1 day and 45 years of age were studied. Serum free T4 was determined by direct equilibrium dialysis, thyrotropin by a sensitive immunoassay, and total T4 by radioimmunoassay. Free T4 concentrations were age dependent and differed significantly (p < 0.001) at 1 to 4 days, 2 to 20 weeks, 5 to 24 months, 2 to 7 years, 8 to 20 years, and 21 to 45 years of age. Corresponding free T4 concentrations (mean +/- SEM) were 48.1 +/- 1.5, 20.3 +/- 0.6, 17.0 +/- 0.4, 19.9 +/- 0.4, and 21.2 +/- 0.4 pmol/L (3.74 +/- 0.12, 1.58 +/- 0.05, 1.32 +/- 0.03, 1.55 +/- 0.03, 1.32 +/- 0.03, and 1.65 +/- 0.03 ng/dl), respectively. Age-related reference ranges were determined. Changes in the relationship between serum thyrotropin and free T4 were complex, indicating age-related changes in pituitary thyroid regulation. No correlation existed between concentrations of free T4 and total T4 after the first 13 days of life (r2 < 0.01). We conclude that (1) pediatric free T4 concentrations and free T4-thyrotropin relationships differ from those in adults and (2) age-related changes in total T4 do not indicate age related changes in either free T4 or thyroid function, after 13 days of age. PMID- 8229523 TI - Uridine diphosphoglucose content of human erythrocytes: assessment by conversion to uridine diphosphoglucuronate. AB - To settle the ongoing controversy regarding differential uridine diphosphoglucose (UDPG) and uridine diphosphogalactose (UDPGal) content of erythrocytes, which may be important in evaluating the metabolic abnormality in patients with galactosemia, we derived a combined enzymatic-high-performance liquid chromatography (HPLC) assay. Uridine diphosphoglucuronate (UDPGA), the unique product of UDPG dehydrogenase activity, was separated and quantified by HPLC in extracts of human erythrocytes. The quantity of UDPGA produced in cell filtrates incubated with the enzyme corresponds to the amount of UDPG directly determined by HPLC. The amount of UDPGA produced was independent of the enzyme purity or activity used. On the other hand, the amounts of UDPG estimated by fluorometric measurement of the production of reduced nicotinamide adenine dinucleotide varied with the enzyme purity and activity. The combined enzymatic-HPLC method confirms the direct determinations of UDPG content of normal erythrocytes. The results indicate that, under appropriate conditions, the fluorometric-based assay will give accurate estimates of UDPG, but the direct HPLC method yields consistent and correct UDPG and UDPGal determinations. PMID- 8229524 TI - Defect in the lipoyl-bearing protein X subunit of the pyruvate dehydrogenase complex in two patients with encephalomyelopathy. AB - Among the many metabolic encephalomyelopathies caused by deficiencies in the pyruvate dehydrogenase complex (PDHC), nearly all involve its E1 subunit. We describe two new familial cases of PDHC deficiency with encephalomyelopathy, chronic lactic acidemia, and a normal E1 subunit of PDHC but deficiency in another component. Activity of PDHC was measured in cultured skin fibroblasts and skeletal muscle, and immunoblot studies were performed on mitochondrial extracts from skin fibroblasts. Spectra of muscle tissue, obtained in vivo with phosphorus 31 nuclear magnetic resonance, were recorded both at rest and with exercise. The PDHC activity was markedly reduced to 10% to 20% of normal values in both cultured skin fibroblasts and skeletal muscle. Immunoblotting of skin fibroblast mitochondrial extracts showed a specific deficiency in the protein X component of PDHC but normal E1, E2, and E3 components. Spectra obtained with 31P nuclear magnetic resonance showed alterations compatible with those found in mitochondrial myopathies. This is the second description of an encephalomyelopathy associated with a specific absence of the lipoyl-containing protein X component, which has a structural role in the formation of a functional PDHC. PMID- 8229525 TI - Major histocompatibility complex class II deficiency: clinical manifestations, immunologic features, and outcome. AB - Major histocompatibility complex class II deficiency (bare lymphocyte syndrome) is a rare primary immunodeficiency disorder characterized by profound defects in human leukocyte antigen class II expression, inconsistent and incomplete expression of human leukocyte antigen class I molecules, and a complete lack of cellular and humoral immune responses to foreign antigens. To define the clinical and immunologic characteristics, outcome, and natural history of major histocompatibility complex class II deficiency, we retrospectively analyzed 30 consecutive patients. Clinical onset occurred in the first year of life, usually involving recurrent bronchopulmonary infections and chronic diarrhea. The clinical course was complicated by viral meningoencephalitis, hepatitis, cholangitis, and various autoimmune phenomena. Prognosis was very poor: the mean age at the time of death was 4 years. The main cause of death was overwhelming viral infection. Recent advances in bone marrow transplantation have raised hopes of curative treatment: 6 of 14 patients who underwent bone marrow transplantation were cured. Long-term survival after human leukocyte antigen-identical and haploidentical bone marrow transplantation seemed to depend primarily on the presence of preexisting viral infections. PMID- 8229526 TI - Hypoprothrombinemia and severe hemorrhage associated with a lupus anticoagulant. AB - We recently encountered a previously healthy 3-year-old girl who had severe bleeding resulting from a severe deficiency of prothrombin. A lupus anticoagulant was identified by several different methods. The patient was successfully treated with glucocorticoids. This rare complication of a lupus anticoagulant should be considered in the differential diagnosis of a previously well child who suddenly has hemorrhage. PMID- 8229527 TI - Membranous nephropathy in two human leukocyte antigen-identical brothers. AB - We describe two Hispanic brothers with membranous nephropathy who had nephrotic syndrome in the first 2 years of life. Secondary causes of membranous nephropathy were excluded by clinical history and laboratory data. The occurrence of membranous nephropathy in these two young brothers, as well as in other familial cases reported to date, suggests an X-linked mode of inheritance. PMID- 8229528 TI - Autosomal dominant transmission of the Pallister-Hall syndrome. AB - We describe a 9-year-old boy and his 34-year-old father with the Pallister-Hall syndrome. The proband had precocious puberty, imperforate anus, postaxial polydactyly, hypospadias, a hypothalamic mass, and a displaced pituitary gland. The father had polydactyly, a hypothalamic mass, and a flattened pituitary gland. We conclude that the most likely cause of the Pallister-Hall syndrome is a mutation in a gene inherited in an autosomal dominant manner. PMID- 8229529 TI - Effect of nitric oxide on the survival rate and incidence of lung injury in newborn lambs with persistent pulmonary hypertension. AB - We previously showed that inhaling nitric oxide (NO) for up to 30 minutes selectively dilates the pulmonary circulation and improves oxygenation in newborn lambs with persistent pulmonary hypertension. In the current study we determined whether inhaling NO for 23 hours increased the survival rate of newborn lambs with persistent pulmonary hypertension, oxidized hemoglobin to methemoglobin, or damaged the lungs. Persistent pulmonary hypertension was created in newborn lambs by ligating the ductus arteriosus 13 days before delivery. Six lambs were randomly selected to breathe NO at 80 parts per million for 23 hours, and 7 control lambs were untreated. Each lamb was delivered at 135 days of gestation (term is 146 days), and the lungs were ventilated at a fraction of inspired oxygen of 0.92. Each of the control lambs died before the end of the study, whereas only one of the NO-treated lambs died (p < or = 0.05). Arterial oxygen tension was greater in the NO-treated lambs by 15 minutes after delivery (63 +/- 17 vs 14 +/- 4 mm Hg). Oxygen tension increased with time in the NO-treated lambs. Inhaled NO increased the concentration of methemoglobin, but this concentration reached a plateau at 3.0% +/- 0.4%. There was evidence of early airway damage in both groups of lambs but no difference between the groups. We conclude that inhaled NO increased survival rates without increasing the incidence of acute lung injury in newborn lambs with persistent pulmonary hypertension. PMID- 8229530 TI - Cost effects of surfactant therapy for neonatal respiratory distress syndrome. AB - OBJECTIVE: To examine the cost effects of a single dose (5 ml/kg) of a protein free synthetic surfactant (Exosurf) as therapy for neonatal respiratory distress syndrome, for both rescue and prophylactic therapy. RESEARCH DESIGN: Nonblinded, randomized clinical trials of both rescue and prophylactic therapy. Regression analyses were used to control for the independent effects of sex, multiple birth, delivery method, birth weight, and surfactant therapy. SETTING: The prophylactic trial was conducted at a university medical center only; the rescue trial also included a tertiary community hospital. PATIENTS: Prophylaxis was administered immediately after birth to 36 infants (38 control subjects) with birth weights between 700 and 1350 gm. Rescue therapy was administered at 4 to 24 hours of age to 53 infants (51 control subjects) with established respiratory distress syndrome and birth weights > or = 650 gm (no upper limit). Infants in the prophylactic trial were not eligible for the rescue trial. RESULTS: For the rescue trial, there was a $16,600 reduction in average hospital costs (p = 0.18), which was larger than the cost of the surfactant ($450 to $900), yielding a probable net savings. For the prophylactic trial, hospital costs were larger for treated infants versus control subjects who weighed less than about 1100 gm at birth and lower for treated infants versus control subjects who weighed more than 1100 gm at birth (p < 0.05). For the prophylactic sample, the result was an average cost per life saved of $71,500. CONCLUSIONS: Single-dose rescue surfactant therapy is probably a cost-effective therapy because it produced a lower mortality rate for the same (and probably lower) expenditure. Single-dose prophylactic therapy for smaller infants (< or = 1350 gm) appeared to yield a reduction in mortality rate for a small additional cost. The use of multiple-dose therapy in infants who do not respond to initial therapy may alter the effects described above to either increase or decrease the observed cost-effectiveness of surfactant therapy. Regardless, surfactant therapy will remain a cost-effective method of reducing mortality rates, relative to other commonly used health care interventions. PMID- 8229531 TI - Arterial oxygen saturation in preterm neonates without respiratory failure. AB - To obtain normal data on arterial oxygen saturation as measured by pulse oximetry (SpO2; Nellcor N200), we obtained 12-hour tape recordings of SpO2, photoplethysmographic waveforms, instantaneous pulse rate, and observations of breathing movements on 55 preterm neonates (25 girls) who had been admitted to one of four special care baby units but had no signs of respiratory distress and were breathing room air at 24 hours of age. Their median gestational age at birth was 35 weeks (range, 30 to 36), and their median age at the time of study 1 day (range, 1 to 7). Median baseline SpO2, measured only during regular breathing, was 99.4% (range, 90.7 to 100; 5th percentile, 95.5). Ten recordings (18%) contained a total of 83 episodes of desaturation (defined as a fall in SpO2 to < or = 80% for > or = 4 seconds). The 95th percentile for desaturation frequency was eight per recording. One infant had 55 episodes of desaturation and thus accounted for two thirds of all episodes observed. Only one of the episodes of desaturation in this infant, and none of those in the other nine infants, had been noted clinically, nor had the abnormally low baseline SpO2 (90.7%) in one infant. Baseline SpO2 in these nondistressed preterm neonates was higher than might be expected, given the SpO2 levels currently recommended for preterm infants with respiratory failure. A minority of infants, however, had a low baseline SpO2 or a high frequency of episodes of desaturation, the potential effects of which remain to be determined. PMID- 8229532 TI - Transient severe respiratory distress mimicking pulmonary hypoplasia in preterm infants. AB - Three preterm infants are described who had respiratory failure immediately after birth. For each, historical factors (extrauterine abdominal pregnancy; ruptured membranes with oligohydramnios for 14 weeks; congenital diaphragmatic hernia) and the initial postnatal course were suggestive of pulmonary hypoplasia. However, with vigorous resuscitation each infant showed rapid improvement in pulmonary status for several hours. Two infants survived with no apparent pulmonary sequelae; the third had almost complete resolution of pulmonary disease but died from gastrointestinal complications. Thus some infants with historical and clinical indicators of severe pulmonary hypoplasia do not have hypoplastic lungs and respond favorably to resuscitation. The causes of the respiratory distress and of its rapid resolution are not known. On the basis of this experience, even infants with historical factors suggesting pulmonary hypoplasia and with postnatal respiratory distress should receive vigorous resuscitation. PMID- 8229533 TI - Effect of early initiation of intravenous lipid administration on the incidence and severity of chronic lung disease in premature infants. AB - OBJECTIVE: To investigate whether intravenous administration of lipid nutrition (Intralipid) within 12 hours of birth to ventilator-dependent premature infants would decrease the incidence or severity, or both, of chronic lung disease. METHOD: We randomly assigned 133 infants to Intralipid or control groups, using two weight strata: 42 Intralipid versus 37 control subjects in the 600 to 800 gm stratum, and 28 Intralipid versus 26 control subjects in the 801 to 1000 gm stratum. The Intralipid group received 20% Intralipid at < 12 hours after birth, starting at a dose of 0.5 gm/kg and increasing to a maximum of 1.5 gm/kg, maintained through day 7. Control infants received no lipid until after day 7. Neither group received enteral feedings until after day 7. RESULTS: No significant differences in mortality rates were present in the total population (23/70 = 32.9% vs 16/63 = 25.4%; p = 0.35, Intralipid vs control); however, the mortality rate increased significantly in 600 to 800 gm infants receiving Intralipid versus the control infants (20/41 = 47.5% vs 9/37 = 24.3%; p = 0.032). No significant differences were found in the number of infants in whom chronic lung disease developed (requiring oxygen for > or = 28 days), proportion requiring oxygen for > or = 60 days, number of survivors without chronic lung disease, or total oxygen and ventilation requirements. However, 600 to 800 gm infants receiving Intralipid had significantly more pulmonary hemorrhage, and greater numbers of infants receiving Intralipid in both weight categories required supplemental oxygen at day 7. The incidence of other complications of prematurity, time required to regain birth weight, and duration of hospital stay did not differ between groups. CONCLUSION: Intralipid administration initiated at < 12 hours after birth failed to protect very low birth weight premature infants from chronic lung disease. Some of the results raise questions about possible deleterious effects of Intralipid when administered early in the first week of life. PMID- 8229534 TI - Neonatal hypoglycemia caused by hypopituitarism in infants with congenital syphilis. AB - Two infants with congenital syphilis and persistent hypoglycemia were found to have hypopituitarism. Hypopituitarism should be recognized as a potential complication of congenital syphilis; affected infants with persistent hypoglycemia should receive a prompt evaluation of pituitary function. PMID- 8229535 TI - Feeding premature infants banked human milk homogenized by ultrasonic treatment. AB - Premature neonates fed ultrasonically homogenized human milk had better weight gain and triceps skin-fold thickness than did a control group given untreated human milk (p < 0.01) and also had lower fat loss during tube feeding (p < 0.01). Ultrasonic homogenization of human milk appears to minimize loss of fat and thus allows better growth of premature infants. PMID- 8229536 TI - A prospective, randomized trial of high-dose intravenous immune globulin G therapy, oral prednisone therapy, and no therapy in childhood acute immune thrombocytopenic purpura. AB - Fifty-three children, aged 7 months to 14.4 years and with typical acute immune thrombocytopenic purpura and platelet counts < or = 20 10(9)/L, were randomly assigned to receive intravenously administered immune globulin G (IVIG), 1 gm/kg per day for 2 consecutive days (n = 19); orally administered prednisone, starting at a dose of 4 mg/kg per day, with tapering and discontinuation of corticosteroids by day 21 (n = 18); or no therapy (n = 16). Both IVIG and prednisone resulted in significantly fewer days with platelet counts < or = 20 x 10(9)/L in comparison with no therapy (median, 1 and 2 days vs 4 days; corresponding ranges, 1 to 20 and 1 to 11 days vs 1 to 132 days; p < 0.01). Reversal of clinically important thrombocytopenia assessed by the number of days taken to achieve a platelet count of > or = 50 x 10(9)/L was significantly faster in children randomly assigned to receive IVIG (median, 2 days; range, 1 to 34 days) than in those receiving prednisone (median, 4 days; range, 2 to 13 days; p < 0.001) or no therapy (median, 16 days; range, 2 to 132 days; p < 0.001). Because the risk of intracranial hemorrhage in children with acute immune thrombocytopenic purpura is highest in the group with severe thrombocytopenia, and appears to be restricted to children with platelet counts < or = 20 x 10(9)/L, these results support the use of IVIG or high doses of prednisone as initial therapy in children with acute immune thrombocytopenic purpura and severe thrombocytopenia (platelet counts < or = 20 x 10(9)/L). PMID- 8229537 TI - A randomized, double-blind, placebo-controlled trial of recombinant erythropoietin in treatment of the anemia of bronchopulmonary dysplasia. AB - Because anemia in patients with bronchopulmonary dysplasia is characterized by inappropriately low serum concentrations of erythropoietin but increased in vitro sensitivity of erythroid progenitors to erythropoietin, we speculated that administration of human recombinant erythropoietin would correct this anemia. Fifteen infants with the anemia of bronchopulmonary dysplasia were randomly assigned to receive erythropoietin or placebo subcutaneously for 10 days. Changes in reticulocyte count, hematocrit, blood lactate concentration, neutrophil count, platelet count, heart rate, oxygen requirement, weight gain, and number of transfusions were assessed. In the 10 erythropoietin recipients (99 +/- 12 days of age), hematocrit values increased from 0.325 +/- 0.006 to 0.381 +/- 0.013 (mean +/- SEM; p < 0.005) and reticulocyte counts from 122 +/- 20 to 446 +/- 48 x 10(3)/microliters (p < 0.005); lactate values remained unchanged. In the five placebo recipients (91 +/- 12 days of age), hematocrits and reticulocyte counts remained unchanged, and lactate values increased from 0.73 +/- 0.14 to 1.34 +/- 0.25 mumol/gm (p < 0.05). During the 30 days after the treatment period, one erythropoietin recipient and four placebo recipients were given transfusions. Other measured variables remained unchanged in both groups. We conclude that erythropoietin is effective in treatment of the anemia of bronchopulmonary dysplasia. PMID- 8229538 TI - Current surgical management of patients with chronic ulcerative colitis. AB - This report consists of a review of an experience with total colectomy with mucosal proctectomy and neorectal reservoir for pediatric patients with ulcerative colitis. The entire experience includes 221 patients, with 73 patients < 21 years of age. Fifty-six consecutive surviving pediatric patients, all > 1 year after operation, have been studied with 100% follow-up evaluation for up to 17 years after operation. Fifty (89%) are leading normal lives with an acceptable stool frequency and complete continence. A number of postoperative complications have been encountered that are of particular concern to the pediatric gastroenterologist. The most distressing finding has been the subsequent diagnosis of Crohn's disease in most of the patients whose postoperative course was unsatisfactory. This underscores the need for a more accurate means of differentiating ulcerative colitis from Crohn's colitis before advising surgery. PMID- 8229539 TI - Malnutrition among weanlings of developing countries: still a problem begging for solutions. PMID- 8229540 TI - Quality-protein maize with a high fat content as a weaning food. AB - Few advances would contribute more to the growth and well-being of infants and children in the developing world than the making available of acceptable staple foods that are easy to grow and process, nutritionally complete enough to serve as the major weaning food, and safe without refrigeration. High-fat quality protein maize (QPM), grown in Mexico, had > 10% of its energy as well-balanced protein and > 15% as fat. When it was fed to six recovering malnourished infants to provide 85-88% of dietary energy, all of 9.8% protein calories, and all of 13.5% fat calories, it was easily consumed and tolerated. In comparison with a fiber-free, casein-based diet, fecal wet and dry weights of children consuming two varieties of high-fat QPM were three to four times greater, and apparent digestibilities of energy were 86 +/- 3 and 86 +/- 4 versus 97 +/- 1%, of carbohydrate 88 +/- 3 and 89 +/- 5 versus 100%, and of nitrogen (N) 77 +/- 4 and 75 +/- 4 versus 89 +/- 2% of intake. High "biological values," 43 +/- 10 and 46 +/- 13 versus 43 +/- 10% of absorbed N, led to comparable apparent N retentions: 33 +/- 8 and 35 +/- 9 versus 38 +/- 9% of intake. Results were similar to those from earlier studies with Peruvian QPM with a typical fat content. Fat absorptions from QPMs were nearly complete, equal to those from separated vegetable oils in the casein diet. Breath H2s were suggestive of significant colonic salvage of incompletely digested maize carbohydrates. PMID- 8229541 TI - Gastrointestinal tolerance, fat absorption, plasma ketone and urinary dicarboxylic acid levels in low-birth-weight infants fed different amounts of medium-chain triglycerides in formula. AB - This study was conducted to evaluate the effect of medium-chain triglycerides (MCT) in a formula for low-birth-weight (LBW) infants on gastrointestinal tolerance, fat absorption, plasma ketone levels, and urinary dicarboxylic acid (DCA) excretion. At the start of enteral feedings, 64 LBW infants (< or = 1500 g) were randomly assigned to one of four experimental formulas. The formulas contained either 0, 17, 34, or 50% of the total fat as MCT oil. The nonfat constituents of all four formulas were the same and identical to Similac Special Care 24 (SCF). Infants were studied from the start of enteral feeding until approximately 7 days after reaching full feeds. Growth and tolerance were assessed in all infants over the entire feeding period. A 48-h balance study was conducted after enteral intake exceeded 100 kcal/kg/day for 3 days. Stool fat, plasma D-(-)-3-hydroxybutyrate (3HB) and carnitine, serum glucose, and urinary DCA levels were determined. Groups did not differ in growth, formula intake, fat absorption (76-84%), serum glucose, or plasma carnitine levels. Gastrointestinal tolerance was excellent and did not differ among groups. Plasma 3HB was significantly different (p < 0.05) only between the 0 and 50% MCT groups, 50 +/- 10 versus 120 +/- 20 microM, respectively. The excretion of urinary DCAs increased with increasing amounts of MCT in the formula. In conclusion, fat absorption and gastrointestinal tolerance were not affected by different MCT levels (0 to 50% of the total fat), but higher levels of plasma 3HB and urinary DCAs were associated with higher levels of MCT in the LBW formulas studied. PMID- 8229542 TI - Newborn rabbit gastric smooth muscle cell culture: EGF and TGF-alpha are potent mitogens. AB - Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) are two in a family of growth-promoting peptides for many gastrointestinal epithelia. This study was designed to assess their mitogenic effect on cultured gastric myocytes and to characterize specific EGF receptors on these cells. Single myocytes were isolated from newborn rabbit gastric fundus and placed into tissue culture. The composition of the culture at confluence as assessed by immunostaining with smooth muscle actin-specific monoclonal antibody (CGA7) was > 95% myocytes. To assess the effect of putative growth factors, freshly isolated myocytes were incubated in Dulbecco's modified Eagle's (DME) medium containing 1% fetal bovine serum in the presence or absence of growth factors. After 6 days, cells were incubated in serum-free medium with [3H]thymidine (1 microCi/ml) in the continued presence or absence of growth factors. After 24 h, EGF and TGF alpha but not insulin-like growth factor I (IGF-I) induced a dose-dependent increase in [3H]thymidine incorporation. Ten nanomolar EGF or TGF-alpha increased [3H]thymidine incorporation more than sixfold over control. EGF was more potent than was TGF-alpha, with apparent median effective dose (ED50) values of 64 +/- 14 pM and 166 +/- 62 pM (p < 0.05), respectively. EGF bound to cultured myocytes with Kd = 7.6 +/- 1.8 nM and Bmax = 27 +/- 11 pmol/mg DNA or 440,000 receptors/cell. TGF-alpha competed for binding at these receptors. Although IGF-I did not stimulate thymidine incorporation, specific high-affinity receptors for IGF-I were detected on gastric myocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229543 TI - Conversion of cholesterol to coprostanol by the intestinal microflora during the first two years of human life. AB - The establishment of the conversion of cholesterol to coprostanol by the intestinal microflora was followed in 25 healthy Swedish children by gas chromatographic analysis of fecal samples taken at 0, 1, 3, 6, 9, 12, 15, 18, 21, and 24 months of age. In 15 children, the microbial conversion of cholesterol to coprostanol started during the second half of the first year of life. During the second year of life, a sequestered distribution of the conversion rate could be distinguished. At the end of the study, 13 of the children were stable high converters, 3 were stable low converters, and 6 were unstable converters while the conversion pattern of three children could not be determined. The initiation of the establishment of conversion was delayed by breast-feeding, and only one child had coprostanol in feces while still being breast-fed. Those children exclusively breast-fed for more than 4 months or weaned after 6 months of age had significantly lower conversion rates at 12 months of age than did the other children (p < 0.05). Once conversion started, previous breast-feeding tended to result in a stable, later high rate of conversion of cholesterol. Between 6 and 15 months of age, the conversion rate increased with age, but after standardization for the period of exposure to foods other than breast milk, no age factor could be identified. PMID- 8229544 TI - Smectite in acute diarrhea in children: a double-blind placebo-controlled clinical trial. AB - Dioctahedral smectite (DS) a natural adsorbent clay capable of adsorbing viruses, bacteria, and other intestinal irritants in vitro, is claimed to possess beneficial "antidiarrheal" properties. This study tested the effect of DS on the duration of diarrhea and the frequency and amount of liquid stools. Ninety well nourished boys, aged 3-24 months, with acute watery diarrhea and mild, moderate, or severe dehydration were included in a randomized double-blind, placebo controlled trial. After initial rehydration, they received DS or placebo (1.5 g freshly dissolved in 50 ml of water, four times daily for 3 days) along with oral rehydration solution (ORS) and adequate feeding. The clinical characteristics of both groups were comparable on admission. Patients in the smectite group had a significantly shorter duration of diarrhea (mean +/- SD, 54 +/- 16 vs. 73 +/- 13 h) and significantly fewer stools (2.6 +/- 0.8 vs. 3 +/- 0.7 on second day; 1.9 +/- 0.7 vs. 2.4 +/- 0.7 on third day; and 11.3 +/- 3.2 vs. 13.8 +/- 3 overall). The amount of liquid stools was not significantly reduced. Weight gain at 24, 48, and 72 h and on recovery was significantly higher in the smectite group despite the comparable fluid and food intake in both groups. These results suggest a beneficial effect of DS in shortening the duration of diarrhea and reducing the frequency of liquid stools in children rehydrated with ORS. PMID- 8229545 TI - Absorption in the isolated bowel segment. AB - An isolated bowel segment (IBS) is a viable mesenteric segment of intestine. This study was undertaken to investigate the effects of mesenteric vascular and neural supply on intestinal absorption. Ten rats were used for leucine absorption and another 10 for glucose absorption experiments. L-leucine, 12 ml of 15.0 mM/L, or 3.0 mM/L of D-glucose solution were recirculated through the IBS (n = 5) and rat jejunum that underwent sham operation (n = 5) at 2.56 ml/min for 90 min. Absorption was expressed as millimoles of leucine or glucose per gram of mucosal protein. The Student's t test was used for statistical analysis; a p < or = 0.05 was considered significant. The net absorption of L-leucine was 36.20 +/- 3.31 mM/g of mucosal protein in the IBS and 15.52 +/- 1.48 mM/g of mucosal protein in the control group (p < 0.001). The net absorption of D-glucose was 3.82 +/- 0.26 mM/g of mucosal protein in the experimental group and 4.34 +/- 0.31 mM/g of mucosal protein in the control group (p = 0.02). This study concludes that absorption of leucine and glucose in the IBS is preserved after mesenteric division. PMID- 8229546 TI - Slow-release 5-aminosalicylic acid therapy in children with small intestinal Crohn's disease. AB - Pharmacologic agents effective in the treatment of Crohn's disease confined to the small intestine are limited. The therapeutic efficacy of oral mesalazine in small bowel inflammation, although theoretically promising, remains unproven. In an open-labeled initial trial, timed-release 5-aminosalicylic acid (5-ASA), administered at a daily dosage of 30.6 +/- 9.0 mg/kg (mean +/- SEM) to children with active Crohn's disease involving the small intestine, was associated with improvement on the Harvey index in six of 12 patients treated for 8.1 +/- 3.9 weeks. In a subsequent prospective, double-blind study 14 children, ages 9.3 to 16.1 years, with active Crohn's disease limited radiologically in the small intestine were randomized to receive either timed-release 5-ASA [50 mg/kg/day (maximum 3 g/day)] or placebo for 8 weeks. Following a 4-week washout period, patients crossed over to receive the other study drug for a further 8 weeks. Six children completed the entire 20-week trial. The van Hees index improved among patients receiving 5-ASA for 8 weeks (delta = -18 +/- 6.4) but deteriorated among patients given placebo (delta = +14 +/- 4.1) (p < 0.05). Mean Crohn's Disease Activity Index (CDAI) decreased marginally after 8 weeks of 5-ASA treatment (delta = -48 +/- 38.2) but not with placebo (delta = -3.0 +/- 7.9) (p = 0.31). Of the eight noncompleters, more patients dropped out of the study because of lack of therapeutic response to placebo (n = 5) than to 5-ASA (n = 2).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229547 TI - Examination of anti-neutrophil cytoplasmic antibodies in childhood inflammatory bowel disease. AB - The detection of anti-neutrophil cytoplasmic antibodies (ANCA), in a perinuclear fluorescence pattern, in the serum of adults with inflammatory bowel disease has recently been described to be sensitive and specific for a diagnosis of ulcerative colitis in comparison to Crohn's disease and other colitides. We have examined the sera of 41 children and adolescents with ulcerative colitis, 27 with Crohn's disease, and a control group for the presence of ANCA. Anti-neutrophil cytoplasmic antibodies were detected in the serum of 27 of 41 patients with ulcerative colitis (66%), five of 27 with Crohn's disease (19%), and in none of our control subjects or patients with functional abdominal pain. Overall, the presence of ANCA was 66% sensitive and 84% specific for a diagnosis of ulcerative colitis when compared to Crohn's disease. There was no relationship between a positive ANCA value and disease activity or other clinical indicators. We conclude that evaluation for the presence of ANCA may be a useful adjunct for the clinical assessment of patients with inflammatory bowel disease. The presence of ANCA in children and adolescents, however, will not definitively distinguish between patients with ulcerative colitis and Crohn's disease. PMID- 8229548 TI - Sonographic findings in celiac disease. AB - In 43 children who had been admitted to our department due to symptoms of malabsorption, sonography of the small bowel was performed before biopsy. Seventeen of the patients were shown to have celiac disease. Twenty-six patients had gastroenteritis, postenteritis syndrome, and enteropathic cow's milk allergy, and one patient had sarcoidosis. At the time of diagnosis, 16 children with celiac disease showed a sonographically abnormal appearance of the small-bowel wall structure. In addition to hyperperistalsis, slight ascites, pericardial fluid, or liver tissue texture changes could be found in some of the patients. Relating the findings of abnormal small-bowel wall structures to the data of the small-bowel biopsy, we found a sensitivity of 94% and a specificity of 88% for sonographic detection of changes related to celiac disease. Our findings indicate that sonographic data can be of great help in exploring children with signs of malabsorption. These data provide a further argument for the decision to perform a biopsy of the small bowel. PMID- 8229549 TI - Epigastric impedance recording for measuring gastric emptying in children: how useful is it? AB - Epigastric impedance recording was used to measure gastric half-emptying time (t1/2) of orange squash in 45 healthy infants and children. The relationship between age, body size, and composition and the impedance increase after gastric filling was investigated, together with the effects on half-emptying time of adding fat, increasing viscosity, and increasing osmolality in the test meal. The mean t1/2 for orange squash was 13.5 min. The increase in impedence per unit volume ingested showed a significant inverse relationship with age (r = -0.62, p < 0.001), and with various anthropometric variables (r ranged from -0.48 to 0.80, p < 0.01). No relationship was observed between t1/2 and age, sex, body size, or composition. Addition of 1.25% triglyceride or a 600-fold increase in viscosity of the test meal were both accompanied by a significant lengthening of t1/2 (p = 0.005 and p < 0.001, respectively), but t1/2 did not alter with a fourfold increase in test meal osmolality. The test was well tolerated by all subjects, but its usefulness is limited by susceptibility to movement artefact, and also the necessity to use simple, liquid test meals. PMID- 8229550 TI - Hepatitis B virus in the colostra of HBeAg-positive carrier mothers. AB - To investigate the amount of hepatitis B virus (HBV) in colostra of hepatitis B e antigen (HBeAg)-positive carrier mothers and the relationship of HBV amount between colostrum and maternal blood, 50 HBeAg-positive carrier mothers were recruited and studied for hepatitis B surface antigen (HBsAg) titer, HBeAg titer, and HBV-DNA concentration in their sera and colostra. According to the presence or absence of seral HBV-DNA determined by dot hybridization, these 50 HBeAg positive carrier mothers could be divided into two groups: group 1 (n = 28, HBV DNA < 0.04 ng/ml), and group 2 (n = 22, HBV-DNA > or = 0.04 ng/ml). The colostral HBsAg and HBeAg titers were both significantly higher in group 2 than in group 1. In addition, both colostral HBsAg and HBeAg titers had positive correlation with each corresponding maternal blood. Although the colostral HBV-DNA was undetectable by dot hybridization, all were positive by polymerase chain reaction with Southern blot hybridization. Because HBV-DNA can be detected in all HBeAg positive carrier mother's milk, it reinforces the necessity of hepatitis B vaccination for the neonates born to these carrier mothers, particularly in countries with a high carrier rate. PMID- 8229551 TI - Successful treatment of fulminant echovirus 11 infection in a neonate by orthotopic liver transplantation. PMID- 8229552 TI - Acute acalculous cholecystitis in infective endocarditis. PMID- 8229553 TI - Association of cytomegalovirus with Menetrier's disease in childhood: report of two new cases with a review of literature. PMID- 8229554 TI - Helicobacter pylori gastric infection and sideropenic refractory anemia. PMID- 8229555 TI - Binding of rotavirus by a 46 kD milk-glycoprotein may prevent gastroenteritis. PMID- 8229556 TI - Transforming growth factor alpha in Menetrier's disease. PMID- 8229557 TI - Tracheostomy: acute and long-term mortality and morbidity in very low birth weight premature infants. AB - Thirty-six very low birth weight premature infants (VLBW-PT) born at 24 to 32 weeks gestation and with birth weights 635 to 1,360 g who had tracheostomies performed for acquired subglottic stenosis or for prolonged mechanical ventilation were followed in relation to acute and long-term mortality and morbidity. Mortality due to the tracheostomy occurred in 4 patients (11%); mortality from all other causes was 25%. Death after hospital discharge was associated with the nonuse of prescribed cardiorespiratory monitors. Complications < 1 week postsurgery occurred in 31% of infants and complications > or = 1 week postsurgery occurred in 64% of infants. Fifty percent of infants required tracheostomy for > 2 years and/or extensive reconstructive surgery of the airway. Parents should be counselled that VLBW-PT infants with a tracheostomy may require extended medical and home care. An effective home care program requires parental training in tracheostomy care, the use of ancillary equipment, and infant cardiopulmonary resuscitation. PMID- 8229558 TI - Surgical resection for pyriform sinus fistula. AB - There has been no established operative procedure for pyriform sinus fistula because of its rarity. Recurrence may occur even after neck exploration. We successfully managed six pediatric cases with this disease without any complications or recurrences. Our experience indicates that exposing the inferior cornu of thyroid cartilage with its covering muscles facilitates resection of the proximal portion of this fistula. PMID- 8229559 TI - Anorectal anomaly: neuropathological changes in the sacral spinal cord. AB - The study was carried out on the bodies of five full-term newborn babies and 10 newborn patients with anorectal anomalies (four of whom had the high deformity, one had the intermediate, and five had the low). Serial transverse sections of the sacral spinal cords at 30 microns were made and stained with Nissle's. It was found that in the medial column of the sacral spinal cord there were two groups of motoneurons: posteriomedian group (group A) and anteriomedian group (group B). The former was found mainly in S3 and S4 and the latter extended from caudal S4 to caudal S2. In six of the 10 deformitive cases the cross-sections of the S5 showed a large diamond-shaped central canal and a thin conus medulla which could be distinguished as the basal and alar laminae; in one case, the lower part of the central canal and the anterior median fissure appeared undeveloped; in one case, the lower part of the central canal was divided into two large canals by a sagittal septum; and in the other two cases, their central canals appeared widened and flattened. Additionally, the average numbers of motoneurons in the group B of the high or intermediate deformity and the low deformity were 13.4% and 61.1%, respectively, of the normal number, and the corresponding numbers of group A were 61.3% and 77.7%, respectively, of the normal number. The observations suggest that sacral myelodysplasia is one of the neuropathological features in anorectal anomaly and may be responsible for the pathological changes in its peripheral nerve system. PMID- 8229560 TI - An immunohistochemical study of neurofilament and microtubule-associated Tau protein in the enteric innervation in Hirschsprung's disease. AB - The distribution and localization of neurofilament (NF) and microtubule associated tau protein (Tau) in the colon from Hirschsprung's disease were examined by immunohistochemistry. Specimens of the normoganglionic, oligoganglionic, and aganglionic segments of colons from nine patients with Hirschsprung's disease were used in this study. Normal colon specimens obtained at the time of colostomy closure from two patients with anorectal malformations were also examined as controls. In normoganglionic segments, anti-NF-H and anti NF-M immunoreactivity appeared within the nerve fibers of both the myenteric and submucosal plexuses. These findings were also observed in the oligoganglionic segments. In the aganglionic segment, hypertrophic nerve fascicules and the nerve fibers in circular muscle were positively stained with anti-NF-H and anti-NF-M antibodies. Anti-Tau staining appeared in the ganglion cell bodies of both myenteric and submucosal plexuses and in nerve fibers distributed among the circular muscles of the normal control colons, and the normoganglionic and oligoganglionic Hirschsprung's specimens. Nerve fibers in the circular muscle layer of aganglionic segments were also stained with anti-Tau serum, although the hypertrophic nerve fascicules in the intermuscular and submucosal layers did not stain. The so-called nerve fascicules distributed in the subserosal layer also did not stain with anti-Tau. These results suggest that Tau may be used as a specific marker to identify ganglion cells and intrinsic nerve fibers in colons affected by Hirschsprung's disease. PMID- 8229561 TI - Lingual thyroglossal duct cyst presenting in infancy. AB - Lingual thyroglossal duct cysts are a rare form of thyroglossal cysts. We present two infants, 10 and 12 weeks of age, with midline intraoral cystic swellings stretching from the base of the tongue to the thyroid cartilage. Complete excision of the cysts by Sistrunk's procedure were carried out. Both infants are well on follow-up, at 6 months postoperatively. Their unique presentation with regard to age, location, and symptomatology is discussed. Lingual thyroglossal duct cysts large enough to cause dysphagia, stridor, respiratory distress, and failure to thrive in infants have not been previously reported in the literature. PMID- 8229562 TI - Central line perforation associated with Staphylococcus epidermidis infection. AB - We present detailed case reviews of four very low birth weight (VLBW) infants in whom central venous silastic catheter perforation was associated with Staphylococcus epidermidis sepsis. The diagnostic and therapeutic dilemma presented by the intracavitary fluid collections occurring in all four of these cases proved to be of clinical interest. Additionally, we propose a model that may account for the etiology of catheter displacement--localized phlebitis as a result of S epidermidis infection with resultant extralumenal migration of the central venous catheter. PMID- 8229563 TI - A safe and cost-effective protocol for the management of esophageal coins in children. AB - Foreign bodies of the esophagus are a common problem in children, with the majority of these foreign bodies being coins. Because there are various methods for managing esophageal coins, we retrospectively reviewed our experience of the past 4 years to establish a safe and cost-effective protocol for their management. From January 1987 to December 1990, 57 children were managed for esophageal coins in our institution. Nineteen children were managed endoscopically under general anesthesia, with a mean patient cost of $1,982 (excluding the surgeon and anesthesiologist charge). Thirty-eight children were managed with Foley balloon extraction or esophageal bougienage in the emergency department with a mean patient cost of $149 (excluding surgeon charge). Both methods were 100% effective, and there were no complications in either group. Rigid criteria were used in selecting patients for balloon extraction and bougienage. These were: (1) duration less than 24 hours; (2) no respiratory distress; and (3) no history of esophageal disease or surgery. Using these criteria, 16 of the 19 patients managed endoscopically could have safely and effectively been managed in the emergency department with a net savings of $1,833 per patient. We conclude that Foley balloon extraction and bougienage of esophageal coins in selected children is both safe and cost-effective. A protocol for managing children with esophageal coins is presented, and the techniques of Foley balloon extraction and esophageal bougienage are reviewed. PMID- 8229564 TI - Evaluation of the right atrial venous oxygen saturation as a physiologic monitor in a neonatal model. AB - Pulmonary artery (PA) mixed venous saturation (SvO2) has become a crucial monitor in the adult intensive care unit, but is not used in neonates because of the difficulty in PA catheterization. We evaluated the possibility of utilizing the right atrial venous oxygen saturation (RAvO2), which is easily accessed in the neonate, as a monitor of the effects of mechanical ventilation and intravascular volume in an animal model selected to be the size of the human neonate. A continuous RAvO2 monitoring catheter was placed into the right atrium of 16 normal rabbits (2.2 to 4.1 kg). Oxygen delivery was manipulated by alterations in peak inspiratory pressure (PIP) (n = 6), positive end-expiratory pressure (PEEP) (n = 6), or by progressive hypovolemia (n = 4). RAvO2 decreased with onset of mechanical ventilation alone from 69% +/- 6% to 61% +/- 5% (P < .01). As the PIP was increased from 12 to 21 cm H2O, the RAvO2 progressively decreased from 59% +/ 4% to 49% +/- 6% (P < .05). As the PEEP was increased from 3 to 9 cm H2O, the RAvO2 progressively decreased from 64% +/- 5% to 33% +/- 16% (P < .01). RAvO2 approached baseline after return to continuous positive airway pressure (CPAP) of 3 cm H2O. Progressive phlebotomy to a total of 10 mL/kg resulted in a decrease in RAvO2 from 70% +/- 6% to 27% +/- 5% (P < .001). Volume resuscitation resulted in an increase in RAvO2 to near baseline. Peripheral arterial oxygen saturation remained at a constant 100% throughout each protocol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229565 TI - Nitrogen balance in patients treated with extracorporeal membrane oxygenation. AB - Extracorporeal membrane oxygenation (ECMO) is a commonly used technique in the management of newborn respiratory failure. However, few studies have addressed the optimal nutritional support of these critically ill neonates. Eleven newborns undergoing ECMO for respiratory failure were studied at various levels of intravenous caloric and nitrogen intake, using nitrogen balance techniques, to assess optimal nutritional support necessary to achieve positive nitrogen balance. Nonprotein nitrogen calories > 60 kcal/kg/d, and nitrogen > 240 mg/kg/d were necessary to achieve positive nitrogen balance, while maximum positive balance was seen with nitrogen intake > 400 mg/kg/d. These data suggest that newborns treated with ECMO can achieve positive nitrogen balance with modest amounts of caloric and nitrogen intake. PMID- 8229566 TI - Long-term results of limited thoracic sympathectomy for palmar hyperhidrosis. AB - Eighteen children (15 females, 3 males) aged 7 to 15 years underwent resection of the thoracic sympathetic chain for severe palmar hyperhidrosis. A localized section of chain immediately below the first thoracic ganglion and including the second thoracic sympathetic ganglion was removed. Patients were followed for 24 to 136 months. All patients had immediate and permanent abolition of palmar hyperhidrosis. There was no mortality, one patient developed intermittent ptosis and myosis, three patients reported compensatory hyperhidrosis and one girl was unhappy with the cosmetic results. We conclude that thoracic sympathectomy is a safe and permanent treatment for severe palmar hyperhidrosis in children. In addition, limited sympathetic resection is associated with a lower incidence of compensatory hyperhidrosis than conventional more radical sympathectomy. PMID- 8229567 TI - Dog bites in children. AB - Dog bites, a common cause of traumatic injury, tend to be a greater source of morbidity and mortality in children than in adults. We evaluated 13 children, who, during a 10-year period, required surgical intervention for dog bites. The greatest cause of morbidity in these children was the penetrating component of the dog bite. Most initial evaluations focused on the crushing component of the bite, but in children with serious dog bites, wounds must be evaluated with respect to both the crushing and penetrating injury components. PMID- 8229568 TI - Abdominal injuries in restrained pediatric passengers. AB - A statewide experience with pediatric abdominal visceral injury in restrained automobile passengers was compiled from the trauma registries of two academic institutions. Retrospective analysis of motor vehicle passenger injuries from 1987 to 1991 included age, sex, mechanism of injury, prehospital care, type of injury, therapeutic interventions, complications, and ultimate outcome. The records of over 2,000 patients evaluated for blunt trauma were reviewed, with 42 children fulfilling the following inclusion criteria: 15 years of age or younger, restrained in an automobile at the time of the accident, and diagnosed with an abdominal injury. Of the 42 patients studied, there were 20 boys and 22 girls; ages ranged from 2 months to 15 years (mean, 7.02 years). Six of 42 patients (14%) required extrication from the vehicle at the scene. Nineteen of 42 patients (45%) sustained belt-related abdominal wall bruising or erythema. The specific blunt visceral injuries noted were as follows: splenic 5, hepatic 5, bowel 6, renal 3, combined 6 (stomach, diaphragm, pancreas, or retroperitoneum). Twenty three children (55%) had abdominal visceral injuries without external seat belt marks. Operative intervention was necessary in seven patients. A delay in diagnosing visceral injury occurred in 4 of 42 (10%) cases. One patient developed abdominal symptoms 72 hours after the accident. Length of hospital stay ranged from 1 to 45 days. Complications occurred in 4 (10%) of patients. There were two deaths due to injuries. Hollow and solid visceral injuries can occur in belted pediatric passengers during vehicular accidents. Both are a source of significant morbidity, and the patient should be evaluated carefully.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229569 TI - Injuries resulting from motorized personal watercraft. AB - To determine if there is a widespread problem with personal watercraft (jet ski) injuries throughout the United States, we reviewed the hospital records of patients who were treated at this institution for injuries incurred while they were operating a motorized personal watercraft or jet ski. All of the patients were under the age of 19 and suffered severe fractures or lacerations. To assess the extent of the problem with these injuries regionally, we collected data from 8 midwestern states for 1989. Sixty-four personal watercraft accidents involving 90 victims were reported in the survey. Fifty-three of 90 patients sustained fractures, lacerations, or head injuries. Seventy-nine of 90 were under age 25, and 24 patients were under 16 years of age. The need for supervision and the potential for serious injury while operating personal watercraft is supported by these findings. PMID- 8229570 TI - Revascularization technique for reduced-size liver transplantation for infants weighing less than 10 kg. AB - Reduced-size liver transplantation has been recognized as a powerful modality in alleviating the global donor shortage in pediatric liver transplantation. We describe, for the first time, a technique for revascularizing reduced-size grafts which has not been patterned after adult revascularization techniques. This revascularization method for reduced-size liver transplantation is particularly suitable for infants weighing < 10 kg. This technique differs from adult revascularization techniques in that the supraceliac aorta is always used as the origin for graft arterialization, and that the anastomoses are always performed in the following order: end-to-side donor celiac artery to supraceliac aorta anastomoses first, followed by the suprahepatic vena caval anastomoses, infrahepatic vena caval anastomoses, and then portal vein anastomoses. Hepatic artery thrombosis in infants weighing < 10 kg has occurred in 4 of 32 nonreduced versus 0 of 21 reduced transplantations (P = .05616, Z test, one tail). Adult revascularization was primarily used in the nonreduced group, whereas our proposed revascularization method was primarily used in the reduced group. We conclude that, for infants weighing < 10 kg receiving reduced grafts, this proposed technique should be used to decrease hepatic artery thrombosis. PMID- 8229571 TI - Ovarian masses in children: a review of 91 cases of malignant and benign masses. AB - Ovarian masses in children are uncommon. We reviewed all cases of ovarian masses presenting to this hospital from 1979 to 1990. Ninety-one patients fulfilled the criteria and had medical records available. All patients were less than 18 years old. Four were diagnosed antenatally. Thirty-four tumors presented prior to 8 years of age and 1 (2.9%) was malignant. Fifty-eight tumors presented after 8 years of age and 18 (33%) were malignant. Seventy-two patients had benign disease and 19 had malignant tumors. Of those with benign disease 22 had simple or epithelial cysts, 25 had teratomas, 13 had torsion with cyst formation, 3 had granulosa cell tumors, and 9 had other less common lesions. Analysis of symptoms could not distinguish between benign and malignant lesions; however, age was less (P < .03) and tumor size smaller (P < .001) in patients with benign lesions. Benign lesions presented at a mean age of 8.8 years. Fifty-four patients had an ultrasound, all were diagnostic: simple mass (14), complex mass (8), or cyst (32). Mean size of the masses was 9.5 x 7.7 cm. Fourteen patients had a contralateral ovarian cyst. The malignant lesions included 14 germ cell tumors (4 endodermal sinus, 4 teratoma, 2 choriocarcinoma, 2 dysgerminoma, 1 embryonal, and 1 mixed), 4 epithelial tumors (1 mucinous cystadenocarcinoma, papillary cystadenocarcinoma, papillary serous cystadenocarcinoma, and endometrioid adenocarcinoma), and one patient with leukemic infiltration (ALL). Germ cell tumors presented at a mean age of 11.8 years. Eight of these patients had an ultrasound and all showed a mass (7) or cyst (1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229572 TI - Anterior sagittal anorectoplasty as a redo operation for imperforate anus. AB - Based on the idea of transsphincteric approach by Pena, we applied anterior sagittal anorectoplasty (ASARP) as a redo operation in 10 patients, who present moderate to severe fecal incontinence postoperatively. Those patients exhibited anal opening located anteriorly to the center of contraction of external sphincter muscles. Operation is begun with making a circumferential skin incision in the mucocutaneous junction around the anal opening, extending posteriorly along the median line to the center of the external sphincter muscles. Upon confirming the vertical muscle and the external sphincter muscles, the rectal tube, being dissected free, is mobilized backward to be placed at the center of the vertical muscles and enclosed by the muscle. After the operation, an improvement was noted in either clinical symptoms or scores, being particularly marked for incontinence and staining scores. Anorectal manometry and barium enema studies also showed a significant improvement. These results provide ample justification of ASARP as a redo operation for imperforate anus being worthwhile trying in properly selected patients with poor anorectal function following the primary operation. PMID- 8229573 TI - A simple and rapid method for staining the enteric ganglia: application for Hirschsprung's disease. AB - A simple and reliable method is described for staining the nerve plexuses of the human gastrointestinal tract in whole-mounts. The staining does not require fixation and consists of a single step of 6 to 10 minutes incubation in a solution of the fluorescent dye 4-(4-diethylaminostyryl)-methylpyridinium iodide. Aganglionic bowel specimens from 6 patients with Hirschsprung's disease were stained by this method and displayed the absence of ganglion cells and an abundance of nerve fibers in the myenteric and submucosal plexuses. This staining may be suitable for the diagnosis of Hirschsprung's disease. PMID- 8229574 TI - Ventral preputial onlay island flap and proximal flip-flap technique for the repair of epispadias: a case report. AB - Use of the ventral preputial island flap technique for repair of epispadias has yielded satisfying functional and cosmetic results. We report a modification of this technique in a patient with proximal penile epispadias without exstrophy. We used the Duckett modification of the onlay island flap using ventral preputial skin and coupled it with a proximal flip-flap of perimeatal skin without division of the urethral plate. This achieved an excellent result. PMID- 8229575 TI - Pyelic benign fibroepithelial polyp in childhood: a case report. AB - Intraluminal masses within the upper urinary tracts are rare, are almost always benign, and most commonly take the form of fibroepithelial polyps within the ureters. Only exceptionally are these lesions found within the renal pelvis and for that reason may cause diagnostic difficulty. We present a new case of fibroepithelial polyp of the renal pelvis treated successfully in our hospital. PMID- 8229576 TI - Idiopathic perforation of the extrahepatic bile duct in infancy: pathogenesis, diagnosis, and management. AB - Three infants with idiopathic perforation of the extrahepatic bile ducts were treated at this hospital during the last 6 years. Two patients presented with jaundice and biliary ascites, with high bilirubin in the ascitic fluid, and one patient with peritonitis. The site of perforation was repaired and abdomen drained in one patient. Simple drainage was performed in two others, with good overall results. Perforation of the bile ducts should be suspected in every neonate and infant with ascites and jaundice. Confirmation is obtained by paracentesis and bilirubin measurement in the ascitic fluid or by combination of sonography and scintigraphy. Simple drainage is the treatment of choice provided the distal obstruction is excluded. PMID- 8229577 TI - Biliary and anorectal atresia. AB - This is a report of a unique case of biliary and rectal atresia. PMID- 8229578 TI - Intraluminal milk curd obstruction in a giant Meckel's diverticulum. PMID- 8229579 TI - Megacystis microcolon intestinal hypoperistalsis syndrome: late sequelae and possible pathogenesis. AB - Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is characterized by hypoperistalsis in the presence of ganglion cells, malrotation, microcolon, bladder distension, and female predilection. We draw attention to the long-term management of a patient with MMIHS and propose a pathogenetic mechanism to account for this syndrome. We propose that the initial event in the pathogenesis of MMIHS is an intramural inflammatory process that affects the gastrointestinal and urinary tracts. This leads to extensive fibrosis which destroys the intestinal neural network, producing hypoperistalsis. The same process causes neuromuscular incoordination in the bladder wall, resulting in irregular bladder contractions against a "closed sphincter" leading to bladder distension. The enlarged bladder then interferes with the rotation of the intestine causing malrotation. PMID- 8229580 TI - Elective surgery for cystic meconium peritonitis: report of two cases. AB - The cases of two infants with prenatally diagnosed cystic meconium peritonitis, who underwent minimal surgical procedure (drainage) first and elective surgery later, are reported. Just after birth tube drains were inserted under local anesthesia with ultrasonographic guidance. They underwent laparotomy, at 16 days in case 1 and at 20 days in case 2. The adhesions were quite easily dissected, ileal atresia was found, and the intestines were safely primarily anastomosed. In cystic meconium peritonitis, radical surgery in the early neonatal period is usually difficult, so many patients require second or third operations. We recommend cyst drainage just after birth and elective surgery later. PMID- 8229581 TI - Imperforate anus, delayed presentation in a 7-year-old girl. AB - Most of the cases of imperforate anus should be discovered at birth, during the initial physical examination. This case represents the status of Third World rural perinatal care, wherein delivery of an infant by a midwife in poverty stricken area of mountainous Jipijapa, Ecuador, failed to reveal the anomaly during the cursory examination. Moreover, the cultural taboos of the Jipijapan agricultural community further delayed the diagnosis. A 7-year-old girl with imperforate anus presenting with a rectovaginal fistula is reported. Specific findings in the case, therapy, and an approach to the diagnosis are discussed. PMID- 8229582 TI - Successful separation of ischiopagus tripus conjoined twins with one twin suffering from brain damage. AB - To our knowledge, until 1988, the year we performed our operation, only six other cases of ischiopagus tripus conjoined twins had been separated in the world. In our case, operated on in October 1988, one twin remained in a vegetative state after an episode of acute and dramatic encephalitis. However, the separation was successfully performed and now, 3 years after the operation, the two continue to live. The particularities of this case and the factors that made the separation successful are reported. PMID- 8229583 TI - Do we use the right entry criteria for extracorporeal membrane oxygenation in congenital diaphragmatic hernia? AB - In a retrospective review we analysed alveolar-arterial oxygen difference (AaDO2) as an entry criterion for extracorporeal membrane oxygenation (ECMO) in neonates with several forms of acute respiratory insufficiency. Although for meconium aspiration syndrome, respiratory distress syndrome, sepsis, and idiopathic pulmonary hypertension of the newborn we found values in accordance with the literature, patients with congenital diaphragmatic hernia (CDH) met 80% mortality criteria with significant lower AaDO2 values. Several patients died before ever reaching usual entry criteria for ECMO, because serious lung deterioration makes AaDO2 values unreliable. Awaiting classical ECMO entry criteria for patients with CDH may at least partially explain the lower survival rate for ECMO in CDH. PMID- 8229584 TI - Intrathoracic silo for fetal diaphragmatic hernia: lung growth and slow reduction of abdominal viscera. AB - Correction of a left congenital diaphragmatic hernia in a human fetus with a large volume of liver in the chest requires reduction of liver and viscera into the abdomen. This can kink the ductus venosus and cause the death of the fetus. Therefore, we have repaired surgically created diaphragmatic hernias in fetal lambs by leaving viscera in the chest wrapped in a silastic chimney. With fetal growth there is a relative reduction of hernia volume over weeks, potentially avoiding kinking the ductus venosus. In four groups of lambs lung size and static respiratory system compliance at birth were compared. Lambs treated by this new technique (silo, n = 7) were compared with lambs that had undergone immediate complete correction with a flat silastic patch in the diaphragm plus an abdominal patch (patch, n = 8), with lambs with uncorrected hernias (n = 6), and with normals (n = 8). There was no significant difference between total lung weights (131 +/- 6 g v 157 +/- 13 g, mean +/- SEM, silo v patch) and lung displacement volumes (142 +/- 7 mL v 162 +/- 14 mL) in either surgically corrected group. Lungs from those corrected by silo were significantly heavier than those with uncorrected herniae (131 +/- 6 g v 56 +/- 5 g, P < .01), but were not as heavy as normal lungs (131 +/- 6 g v 257 +/- 16 g, P < .01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229585 TI - Congenital cystic adenomatoid malformation of the lung: prenatal management and prognosis. AB - Thirty-two cases of congenital cystic adenomatoid malformation of the lung diagnosed antenatally are reported. Antenatal diagnosis has made it possible to document the progress of the condition in utero and the postnatal prognosis. It has been possible to advise on termination and intrauterine intervention, to counsel the parents, and to plan arrangements for delivery and postnatal care among obstetricians, neonatologists, and pediatric surgeons. According to Stocker's classification there were 12 cases of type I, 15 of type II, and 5 of type III. Five pregnancies were terminated. Antenatal drainage of a cyst was performed in four patients with two survivors. Thirteen babies showed relative regression of the cyst as pregnancy progressed. After delivery the extent of the cystic malformation was assessed by chest x-rays and computed tomography scanning in 25 and angiography in 6. Treatment consisted of observation in 4, embolization in 2, operation as an emergency in 3, and electively around 4 months in 15. PMID- 8229586 TI - The clinical application of the Malone antegrade colonic enema. AB - Continent, catheterizable enterostomies (Malone stomas) have been formed in 25 patients. A reversed appendicocecostomy was used for the majority, although experience has led to modifications of this procedure. Antegrade enemas are regularly administered via these stomas in the treatment of fecal incontinence and intractable constipation. There have been only two important surgical complications, and the technique has proved to be highly successful. Follow-up of the patients has shown that individual requirements can be met with variations in the administration of the enemas. PMID- 8229587 TI - Conservative surgery of bladder-prostate rhabdomyosarcoma in children: results after long-term follow-up. AB - Between 1968 and 1982, 10 children (7 boys and 3 girls) with a mean age of 3.2 years affected by bladder (8 cases) and prostate (2 cases) rhabdomyosarcoma (RMS) -all histologically embryonal--were seen. Three (1 with bone metastases at diagnosis and 2 with stage III tumor) underwent radical surgery; all died because of progression of the disease. Seven children were treated with conservative surgery: 3 solid type bladder tumors were removed together with a narrow strip of apparently healthy bladder wall, sparing 30% to 70% of the bladder; 3 botryoid type bladder tumors were removed bluntly and implantation sites fulgurated; in 1 prostate tumor osmotic cytolysis of the neoplastic mass was achieved with multiple injections of hypertonic saline solution (NaCl 30%). Surgery was followed by chemotherapy with a vincristine actinomycin D and cyclophosphamide protocol and radiotherapy (20 to 39.5 Gy). Six of the seven are alive and disease free with a normal functional bladder after 10 to 21 years. The validity of a primary conservative surgical approach with preservation of the bladder, followed by adjuvant therapy, is emphasized. PMID- 8229588 TI - Bladder management in children with genitourinary sarcoma. AB - Between 1977 and 1991, 14 patients were treated for genitourinary sarcoma (mean age, 7.4 years). The primary site was bladder in six patients, prostate in five, and vagina in three. Histological study showed embryonal rhabdomyosarcoma in 12 and leiomyosarcoma in 2 children. Initial therapy included biopsy followed by chemotherapy in all patients. Subsequently, five patients had anterior exenteration, four underwent partial cystectomy, and one patient had a radical prostatectomy; four patients were treated with chemotherapy and external beam pelvic irradiation (4,000 to 6,000 rads) alone. Overall survival for the group is 100% (follow-up 6 to 168 months). Two of four patients undergoing partial cystectomy had bladder augmentation at the time of surgery. All patients having partial cystectomy had negative surgical margins, are tumor free, and have volitional voiding. Two of four patients (50%) initially treated with chemotherapy and radiotherapy alone have had significant bladder deterioration requiring bladder reconstruction. There has been an evolution toward less radical, initial surgical intervention in pediatric genitourinary sarcoma; however, surgical resection continues to be the primary curative modality. Partial cystectomy with or without primary reconstruction may be preferable to exenteration for selected patients. Primary reconstruction at the time of partial cystectomy leaves a functional bladder and excellent long-term results. Children treated with chemotherapy and radiotherapy protocols alone must be monitored for late bladder deterioration. PMID- 8229589 TI - The desmoplastic round cell tumor: a new solid tumor of childhood. AB - Three patients with a new, pathologically distinct solid tumor of childhood have been treated recently. The disease is characterized by male predominance, adolescent onset, an extensive abdominal primary tumor, and aggressive metastases to regional lymph nodes, liver, and lung. Two patients presented with vague abdominal pain and the third with testicular pain. All three noted fatigue and malaise of less than two months' duration with minimal associated weight loss. Computed tomography (CT) scans of the abdomen and chest were obtained for initial preoperative staging, and then all three underwent surgical exploration. Widespread disease was found in each case. In no instance was complete tumor extirpation possible because of extensive peritoneal spread and lymphatic and hepatic metastases. Histologically, all three tumors consisted of round blue cells with a dense desmoplastic reaction and focal rhabdoid features. Immunohistochemical markers for epithelial, neural, and muscle elements were positive. Aggressive multidrug chemotherapeutic regimens were used in each case, and all three patients are alive and well but with known residual disease. We conclude that in cases of the desmoplastic round cell tumor of childhood, CT scans underestimate the extent of disease, and exploratory laparotomy is necessary for diagnosis and appropriate staging. Surgery is usually palliative because of extensive spread. Awareness of this newly recognized aggressive solid tumor of childhood is essential to define its natural history and guide the development of effective multidisciplinary therapeutic regimens. PMID- 8229590 TI - Is contralateral exploration necessary in infants with unilateral inguinal hernia? AB - Between 1973 and 1985, 165 infants (aged 1 week to 6 months) underwent unilateral inguinal herniotomy at our hospital. An attempt was made to trace these 165 children. It proved possible to contact and examine 116 children (104 boys and 12 girls). Age at follow-up ranged from 5 to 17 years. Parents were asked whether their children had attended any hospital for the treatment of contralateral hernia. All children were examined for the evidence of contralateral hernia. Boys were also examined for the position and size of the testis. Testicular volume was assessed with the help of Prader's Orchidometer. Twelve children (10.3%), 11 boys and 1 girl, subsequently developed contralateral inguinal hernia. The mean time interval between initial hernia operation and subsequent development of contralateral hernia was 164 days (range, 7 days to 18 months). Diminished size of testes was observed on the side of the operation in six patients and one patient had complete testicular atrophy. Three boys had a testis in the groin, presumably hitched up at operation and all three required orchidopexy. In view of the relatively low incidence of contralateral hernia and increased risk of damage to testes, we feel that routine contralateral exploration is not justified. PMID- 8229591 TI - Management of primary megaureter in infancy. AB - The natural history of primary megaureter was evaluated in 26 infants (20 boys, 6 girls) with 38 nonrefluxing primary megaureters. Prenatal ultrasound diagnosis had been made in 21 infants. In all patients, primary megaureter was associated with pelvicaliceal dilatation. Two patients with urinary tract infection and two with agenesis of the contralateral kidney underwent surgical treatment. Asymptomatic infants with good differential renal function on DTPA scanning were managed conservatively regardless of their postfurosemide drainage curve. One of these infants later developed a slight deterioration in function and was submitted to surgery. One patient was lost to follow-up. Twenty patients, after a period ranging from 4 months to 8 years, showed partial or complete regression of ureteral dilatation with stable good renal function. In conclusion, many asymptomatic primary megaureters in infancy improve spontaneously and do not require surgical treatment. PMID- 8229592 TI - A long-term follow-up of surgically treated vesicoureteric reflux in girls. AB - Ninety-six girls with surgically treated vesicoureteric reflux (VUR) operated on between 1970 and 1975 were studied to determine their post mature growth (height and weight); the incidence of hypertension; the incidence, progression, and development of new renal scars; the incidence of urinary infection; and the outcome of any pregnancies. The average age at follow-up was 20 years 2 months (range, 16 years to 25 years). The final "post mature" height was significantly greater than that of a group of controls from all regions of Great Britain; the incidence of hypertension was 4.5% and the incidence of urinary infections was 37.6%. The majority of subsequent pregnancies were uneventful but urinary infections occurred in 36.4% and 3 of the 20 infants born had VUR. PMID- 8229593 TI - Limitations of extracorporeal shock wave lithotripsy for urinary tract calculi in young children. AB - Despite success rates with a variety of urinary tract calculi, there is growing concern that extracorporeal shock wave lithotripsy (ESWL) has limitations and that its role needs to be redefined. We report the outcome of 28 consecutive children (age range, 6.5 months to 7 years; mean, 3.6 years) with urinary calculus disease, treated over a 5-year period. Thirteen patients had ESWL monotherapy, and 8 achieved stone clearance. The other 5 children in the ESWL monotherapy group, all with multiple calculi, required surgery to render them stone free. A further 14 patients (6 staghorn calculi, 6 multiple calculi, 1 solitary renal, and 1 child with multiple bladder calculi) were considered unsuitable for ESWL and had primary surgery. Twelve of those 14 were cleared by open surgery, one had residual fragments successfully treated by ESWL, and one still awaits adjuvant ESWL. One child had a solitary renal calculus (5 mm) which passed spontaneously. This study demonstrates that ESWL monotherapy cleared stones in only 8 of 28 patients and clearance in a further 6 was achieved with surgery. Surgery will continue to play an important role in the management of paediatric urolithiasis for large staghorn, multiple urinary tract calculi and lithotripsy failures. PMID- 8229594 TI - Liver transplantation in babies and children with extrahepatic biliary atresia. AB - Orthotopic liver transplantation (OLT) is a life-saving procedure for end-stage liver failure. We reviewed 39 children (24 girls, 15 boys) who received OLT for biliary atresia from 1987 to 1991. Twenty had unsuccessful portoenterostomy, 6 were referred too late for a drainage operation, and the remaining 13 achieved bile drainage but developed portal hypertension. At transplant 37 had decompensated liver disease with varices (28), ascites (24), encephalopathy (17), and gastrointestinal bleeding (12). The median weight and age at transplant were 8 kg and 12.6 months, respectively. The median waiting time was 65 days. Forty eight grafts (30 reduced and 18 whole) were performed; graft loss was 33% and 27%, respectively. Of the 30 segmental grafts, 15 were reduced conserving the left lateral segment and hepatic vein (Brisbane technique)--13 were from the left lobe and 2 from the right lobe. The overall subject survival rate is 72%. Eleven deaths occurred: primary nonfunction (3), sepsis (3), perioperative bleed (3), and other causes (2). Early complications included: hepatic artery thrombosis (5), hepatic vein thrombosis (2), bowel perforation (3), biliary leak (3), and acute rejection (8). Later complications were chronic rejection (4) and biliary stricture requiring reconstruction (3). Follow-up at 12 months confirms good quality of life for both child and family with catch up growth and normal development. Technical advances in reduction hepatectomy have allowed us to treat small babies under 1 year with an urgent requirement for OLT, with comparable results to those obtained with whole grafts. In conclusion, in the future size and age need not be a contraindication to OLT in children with biliary atresia. PMID- 8229595 TI - Is age less than 1 year a high-risk category for orthotopic liver transplantation? AB - The aim of this study was to determine if age less than 1 year is a high risk group for orthotopic liver transplantation (OLT). Retrospective analysis was done of patients with liver failure who received OLT. Comparison was made between patients aged < 1 year and > 1 year with regard to survival, allograft survival, hepatic artery thrombosis, and medical status at OLT. Between January 1, 1987 and September 30, 1991, 46 children received OLT. Fifteen (35%) were < 1 year (average age, 7.93 months). Survival in children < 1 year was 80% and children > 1 year was 91%. Allograft survival in children < 1 year was 57% (21 allografts required for 12 survivors) and 78% in children > 1 year (37 allografts required for 29 survivors). Retransplantation was required in 5 of 15 children < 1 year (33%) and in 5 of 29 children (17%) > 1 year. Medical status in children < 1 year was similar to medical status in children > 1 year at the time of transplant. Children with chronic stable liver disease represented 60% of children < 1 year and 60.1% of children > 1 year. Children requiring hospitalization represented 26% of children < 1 year and 29% of children > 1 year. Children in intensive care represented 13% of children < 1 year and 11% of children > 1 year. Survival for all status groups was similar. Hepatic artery thrombosis occurred in one child < 1 year and in 2 children > 1 year. No statistical difference (chi 2 analysis) was found by age between the categories evaluated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229596 TI - Liver transplantation in small babies. AB - Pediatric liver transplantation is an effective treatment for end-stage liver disease with 1- and 5-year survivals approaching 90% and 70%, respectively. Survival is influenced by the recipient's age, weight, primary disease, vascular malformations, and nutritional status. Younger patients weighing less than 13 kg are considered to be a high-risk group. The aim of this article is to evaluate the impact of this group of patients on the overall results of our pediatric liver transplant program. From January 1986 through January 1992 we performed 76 liver transplants in 59 pediatric patients. Sixteen received a second graft and a third was required in one. Fourteen patients weighed less than 13 kg (mean, 11 kg; range, 6 to 13 kg). Their mean age was 12 months, with a range of 8 to 36 months. Indications for transplantation were: biliary atresia (9), Byler's disease (1), tyrosinemia (3), and alpha 1-antitrypsin deficiency (1). The incidence of rejection in this group (52%) was not significantly different from that in other patients (61%). Ten episodes of acute rejection required only steroids: in one monoclonal antibodies were added. Five patients had a new graft implanted, four for hepatic artery thrombosis and one for primary liver nonfunction. Nine patients are alive (64%) with the follow-up time ranging from 2 to 56 months (mean, 31). Five patients died of multiorgan failure (3), portal vein thrombosis (1), and primary liver nonfunction (1). Four-year graft and patient survival rates were 47% and 64%, respectively. Small babies are a high risk group in a pediatric liver transplant program.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229597 TI - Constipation is a major functional complication after internal sphincter-saving posterior sagittal anorectoplasty for high and intermediate anorectal malformations. AB - Anorectal function was clinically, manometrically, and radiologically followed-up in 40 patients who underwent an internal sphincter-saving posterior sagittal anorectoplasty in 1984 to 1989. The functioning internal sphincter was manometrically verified in 83% (33/40) of the patients. Soiling related to sphincter insufficiency was found in only 4 of the 33 patients with a functioning internal sphincter; 5 of the 7 patients without an internal sphincter had soiling. Symptomatic constipation was found in 73% (24/33) of the patients with a functioning internal sphincter, but in only 28% (2/7) of the patients without this structure. Constipation was not associated with a stenotic anal outlet in any patient. On anorectal manometry, there was no statistical difference in basal and pressures and internal sphincter reflex threshold values between constipated and nonconstipated patients. Histologically, ganglion cells were found in the proximal anal canal in all cases. The radiological size of the rectal pouch before closure of the protecting colostomy had a positive correlation with the severity of constipation. Medical treatment with diet and bulk-laxatives (13 patients) or with stimulant laxatives and occasional enemas (11 patients) successfully relieved constipation in 24 patients. In 2 patients, the symptoms were refractory to medical treatment and a resection of the megarectum was required. In conclusion, the preservation of the internal sphincter in patients with high or intermediate anorectal anomalies gives a good fecal continence outcome, but is associated with a high incidence of symptomatic constipation. PMID- 8229598 TI - Adaptation in short-bowel syndrome: reassessing old limits. AB - The improving survival of patients with severe short-bowel syndrome along with the advent of successful intestinal transplantation have accentuated the need to answer two questions. (1) Is there an intestinal length below which adaptation to full enteral nutrition can not be expected to occur? (2) How much time is necessary to complete intestinal adaptation? We reviewed the outcome of 21 infants with less than 50 cm of small intestine to answer these questions. Patients were divided into three groups based on intestinal length, regardless of ileocecal valve status: group I, < 10 cm (n = 3); group II, 10 to 30 cm (n = 11); and group III, 30 to 50 cm (n = 7). Data were collected to assess survival, incidence of adaptation, time to adaptation, and causes of mortality. Infants in group I did not achieve intestinal adaptation to full enteral nutrition. One survived and 2 died, one from varicella pneumonia and the other after intestinal transplantation. Eight of the 11 (73%) patients in group II survived and 5 of 8 (63%) survivors achieved full intestinal adaptation after a mean interval of 320 days (range, 148 to 506 days) on parenteral nutrition. Six of the seven patients (86%) in group III survived and all survivors (100%) achieved complete enteral adaptation after an average of 376 days (range, 58 to 727 days). The overall survival was 71% (15/21), but survival in patients with > 10 cm was 78%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229599 TI - Trauma scores, accident deformity codes, and car restraints in children. AB - The importance and effectiveness of the appropriate use of automobile restraints by young children has been emphasized in several studies. Once the child has entered the emergency care system, however, restraint use may not be the best predictor of injury severity. This study was undertaken to investigate the relationship of restraint status to morbidity and mortality in children examined in a hospital emergency facility following involvement in a motor vehicle crash (MVC). The emergency room charts of 101 children under 18 years of age, who were victims of MVCs, were reviewed and the following trauma scores were calculated: Glascow Coma Scale, Pediatric Trauma Score, Revised Trauma Score, Injury Severity Score, and Maximum Abbreviated Injury Score. In addition, the Traffic Accident Damage (TAD) score, an estimate of crash severity determined by the police at the accident scene, was recorded. The patients were age stratified as follows: 0 to 4 years (n = 24), 5 to 11 years (n = 29), and 12 to 17 years (n = 48). Fifty patients were appropriately restrained at the time of the crash. There was a significant correlation between mean trauma scores and mean TAD codes (P < .05). There were no significant differences in mean trauma scores between improperly restrained (n = 11) and unrestrained (n = 40) children (P > .05) across all age groups, and these children were grouped together as "unrestrained" in further analyses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229600 TI - The premature infant with esophageal atresia and respiratory distress syndrome. PMID- 8229601 TI - Pilomatrixomas in two children. PMID- 8229602 TI - Recurrent necrotizing enterocolitis. AB - In the decade 1981 to 1991, 16 infants developed recurrent necrotizing enterocolitis (NEC). They comprised 12 (6%) of 196 neonates referred for further management of NEC and four others referred with major congenital anomalies. Their median gestational age was 32 weeks (range, 27 to 40), median birth weight was 1,260 g (range, 790 to 3,230), and the sex distribution was equal. Recurrent NEC occurred after a median interval of 37 days (range, 11 to 163) from the onset of the initial episode. All but one of the 16 infants were either premature (n = 10) or mature with major congenital anomalies (n = 5). Nine patients had previously undergone surgery for NEC. Medical treatment was successful in 11 patients with recurrent NEC. Two infants died, both of liver failure related to parenteral nutrition; one of whom had suffered four separate episodes of NEC and was found to have a superior mesenteric artery occlusion. There was no consistent association between recurrent NEC and the type or timing of enteral feeds or the anatomical site or method of management of the original attack. The mortality of 12.5% was similar to that for primary NEC. PMID- 8229603 TI - Necrotizing enterocolitis complicating the management of posthemorrhagic hydrocephalus. AB - The case notes of all newborn infants with posthemorrhagic hydrocephalus (PHH) operated over the past 10 years at our institution were reviewed to establish the incidence and the effect of necrotizing enterocolitis (NEC) on morbidity and mortality following cerebrospinal fluid (CSF) shunting. Thirteen neonates had both PHH and NEC (group A); 7 of these patients were initially treated by ventriculoatrial (VA) shunt and six by ventriculoperitoneal (VP) shunt. Seventy five patients had PHH alone (group B); all were treated by VP shunt. Eight patients in group A required an abdominal operation for NEC. The two groups were comparable for birth weight, gestational age, and other complications of prematurity. Episodes of shunt malfunction (infection and/or obstruction) and deaths occurring within 12 months from shunt insertion, in the two groups were compared. Shunt malfunction was more frequent in group A (72%) than in group B (27%) (P < .001). Shunt infection was observed in 39% of group A patients versus 14% in group B (P = .03). Distal shunt obstructions occurred in 28% of group A patients and only 3% of group B patients (P = .001). There were more deaths in group A (62% v 9%; P < .001). Thirty-one percent of group A patients and 4% of group B patients died following shunt complications (P = .006). In group A, there was no significant difference in mortality and shunt malfunction between patients with VA or VP shunts. The method of treatment and the stage of NEC did not influence morbidity and mortality after internal drainage for PHH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229604 TI - Bridging of esophageal defects with lyophilized dura mater: an experimental study. AB - The aim of this study was to investigate the use of a prosthetic biologic material-lyophilized dura mater (Lyodura) in patching esophageal defects in dogs, having in view its potential use in bridging long gap congenital esophageal atresia and patching acquired esophageal defects. To follow the incorporation process, 20 mongrel dogs had full-thickness, 6 cm2 cervical esophageal defects, patched with Lyodura. The dogs were sacrificed at different postoperative periods and the patched esophagus removed for gross and microscopic examination. In a second group of 10 dogs, a segment of the esophagus was excised and replaced by 3 cm in length and 2 cm in diameter, Lyodura tubes. In this group, prolonged follow up was undertaken including radiologic, endoscopic, and histological assessment. Special attention was given to the swallowing function and to growth and development. This study shows that: (1) the area of prosthetic replacement was characterized by a narrowing of the esophageal wall and the histopathologic study showed that the epithelialization process was complete in about 2 months; and (2) muscle reconstruction did not take place at the patched areas. We conclude that, pending further studies and improvement, lyophilized dura mater can be considered as a successful alternative for bridging esophageal defects. PMID- 8229605 TI - Development of a technique for jejunal interposition in long-gap esophageal atresia. AB - Primary anastomosis is the treatment of choice in esophageal atresia. There remains a small number of infants in whom establishment of primary esophageal continuity fails or is unrealistic due to a long gap. Esophageal reconstruction then necessitates an interposition/transposition procedure. The most widely used tissues are colon and stomach but each has significant limitations. Jejunum is theoretically the ideal esophageal substitute being of appropriate diameter and having good peristaltic activity. Its use in the past has been limited by the precarious blood supply and restricted length which result from the short mesenteric pedicle. We have attempted to resolve these limitations by using a microvascular anastomosis to augment the blood supply to the proximal jejunum in a series of 5 cases (2 pure atresias, 1 esophageal atresia with proximal fistula, and 2 atresias with distal fistula). In a sixth case (atresia with distal fistula) a free jejunal graft was used. In one case initial surgery was confined to cervical esophagostomy and feeding gastrostomy, in the remaining 5 interposition was necessitated by failure of a primary repair. The age at surgery ranged from 8 to 16 months. The development of the technique and outcome in each patient is described. We conclude that a free jejunal graft is preferable to augmenting the native blood supply and intend to continue with this latter technique. PMID- 8229606 TI - Airway obstruction and gastroesophageal reflux: an experimental study on the pathogenesis of this association. AB - Gastroesophageal reflux (GER) frequently complicates the clinical course of children suffering from conditions leading to upper airway obstruction (UAO) (choanal atresia, tracheomalacia, esophageal atresia, vascular rings, etc). In an attempt to explore whether partial airway obstruction causes changes in the normal thoracoabdominal pressure gradients, we measured end-inspiratory intrathoracic and intraabdominal pressures in anesthetized rats under spontaneous breathing conditions, after tracheostomy, and under upper airway obstruction induced by tracheal intubation with three progressively narrower cannulae (inner diameters 1.0 mm, 0.5 mm, and 0.2 mm). We also measured the lower esophageal sphincter pressure (LESP) and length (LESL) and calculated the thoracoabdominal end-inspiratory pressure gradient (TAEIPG). Neither LESP nor LESL changed significantly before or after maximal tracheal obstruction (14.3 +/- 6.2 v 18 +/- 7.6 cm H2O [P > .05] and 0.34 +/- 0.09 v 0.41 +/- 0.1 cm H2O [P > .05] respectively) but TAEIPG significantly increased from 5.58 +/- 1.34 cm H2O to 17.62 +/- 4.27 cm H2O (P < .01) under the same conditions, mainly as a result of progressively increasing negative intrathoracic pressures during inspiration. These experiments prove that the powerful thoracoabdominal pressure gradients developed after partial UAO may contribute to the pathogenesis of GER by overcoming the antireflux barrier function. This study points out the convenience of routinely screening for GER all children with airway obstructive conditions, bearing in mind that the reestablishment of normal respiratory conditions should be the primary goal of treatment. PMID- 8229607 TI - An association of pulmonary hypoplasia with unilateral agenesis of the diaphragm. AB - During a period of 5 years, 33 newborns with congenital diaphragmatic hernia were treated. Three groups presenting with respiratory distress in the delivery room were identified. These included 8 newborns with agenesis (group 1) and 4 newborns with nonagenesis (group 2), all of whom died. There were 19 nonagenesis survivors (group 3), giving an overall survival rate of 61%. Two newborns who presented beyond 6 hours of life were excluded. No one specific arterial blood gas value or ventilation parameter obtained preoperatively could predict survival. Postmortem right and left lung weights, lung/body weight ratio, and radial alveolar counts demonstrate that agenesis is a unique subgroup with profound pulmonary hypoplasia and a dismal prognosis. PMID- 8229608 TI - Intravenous immunoglobulin therapy in neonatal immune hemolytic jaundice. AB - Nine cases of newborn patients who developed hyperbilirubinemia due to blood group incompatibility and were treated with high dose (1 gram/Kg) intravenous immunoglobulin (IVIG) are described. In 7 of the 9 patients the rise of bilirubin level was attenuated and exchange transfusion was not required following treatment with IVIG. Of the two patients who did require an exchange transfusion despite IVIG treatment, one had the exchange performed immediately following the IVIG infusion, allowing no time for observation of the therapy effect. Our results suggest that IVIG administration may be efficacious in the treatment of immune hemolytic hyperbilirubinemia of the newborn but further studies are required to validate the efficacy of this treatment. PMID- 8229609 TI - Treatment of neonatal hyperbilirubinemia with repetitive oral activated charcoal as an adjunct to phototherapy. AB - The efficacy of multiple dose oral activated charcoal (OAC) therapy for neonatal hyperbilirubinemia was prospectively studied in 30 jaundiced newborns receiving phototherapy, randomly assigned to a study group (n = 14) or control group (n = 16). The study group received OAC before meals with a total amount of 8.5 +/- 0.85 gms (M +/- SEM). Serum bilirubin levels upon initiation of phototherapy were (M +/- SEM) 265 +/- 8 and 253 +/- 4 mumol/L respectively. After 24 hours there was no significant decrease in serum bilirubin levels in the control group (M +/- SEM = 240 +/- 8 mumol/L) but bilirubin levels of the study group decreased (M +/- SEM = 235 +/- 7 mumol/L, p < 0.02). At 48 hours serum bilirubin levels were significantly lower than baseline values in both groups. However, the decline in bilirubin levels in the study group (M +/- SEM = 56 +/- 10 mumol/L) was greater than that of the controls (M +/- SEM = 21 +/- 10 mumol/L p < 0.02). Oral activated charcoal seems to be an effective adjunct to phototherapy in the treatment of neonatal hyperbilirubinemia. PMID- 8229610 TI - Infant neurodevelopmental outcome in pregnancies complicated by gestational hypertension and intra-uterine growth retardation. AB - The neonatal outcome of 78 consecutive singleton pregnancies complicated by intrauterine growth retardation (IUGR) and gestational hypertension were compared with the outcome of 78 adequately matched pregnancies complicated by idiopathic IUGR. The rate of low (< 5) 1-minute Apgar scores was higher in infants born to hypertensive mothers (12.8% vs 2.6% p = .035). No differences in the prevalence of other perinatal factors such as acidosis, respiratory distress syndrome, hypoglycemia, pneumothorax, bronchopulmonary dysplasia, intracranial hemorrhage, requirement for assisted ventilation or survival were found between cases and controls. After two years' follow-up, the rate of major neurological neonatal handicaps, was 2.8% in the cases and 1.4% in the controls (p = 0.56). Mild neurodevelopmental abnormalities were more frequent in infants born to hypertensive mothers (14.3% vs 2.9% p = .025). After adjustment by multiple logistic regression, to eliminate the effect of confounding factors, the probability of normal neurodevelopmental outcome was reduced by 82% in infants born to hypertensive mothers as compared to controls (Odds Ratio = 0.18; 95% confidence interval 0.05 to 0.82 p = .028). These findings suggest that pregnancies complicated by IUGR and gestational hypertension are associated with a high prevalence of subsequent neurodevelopmental problems among infants. PMID- 8229611 TI - Vitamin A levels and feeding practice in neonates with and without chronic lung disease. AB - Infants with chronic lung disease (CLD) have been demonstrated to have low vitamin A levels. The aim of this study was to assess factors which influenced this association. Vitamin A levels of infants born at less than 32 weeks gestational age with and without CLD were determined and related to feeding practice. The infants with CLD had statistically significantly lower vitamin A levels on days 21 to 30 and days 31 to 40. From day 11 onwards, the median Vitamin A levels were less than 60% of the minimum acceptable level for a healthy child or adult. Infants with CLD required significantly longer mechanical ventilation and parenteral nutrition than controls and there was a significant positive correlation between duration of mechanical ventilation and parenteral nutrition (r = 0.83). Infants with CLD received significantly less vitamin A supplementation than the infants without CLD. Our results thus suggest that feeding practice explains the association of CLD and vitamin A deficiency in preterm infants. PMID- 8229612 TI - Transport of hypoxanthine from plasma to cerebrospinal fluid and vitreous humor in newborn pigs. AB - To determine whether an elevated level of hypoxanthine in cerebrospinal fluid or vitreous humor might reflect a high plasma hypoxanthine concentration, or whether it necessarily represents local tissue hypoxia, we infused hypoxanthine intravenously to normoxemic and normotensive piglets (n = 6). Hypoxanthine was measured in different body fluids using HPLC. During the 8 hours of infusion hypoxanthine increased in plasma (from 30 +/- 6 mumol/l (mean +/- SD) before the infusion to 68 +/- 20 mumol/l at the end of the infusion, p < 0.01), cerebrospinal fluid (CSF) (19 +/- 8 to 43 +/- 9 mumol/l, p < 0.05) and vitreous humor (15 +/- 5 to 30 +/- 6 mumol/l, p < 0.05). After infusion, hypoxanthine values in all three fluids were similar to those seen in pigs after severe hypoxia. Hypoxanthine in vitreous humor and plasma were significantly correlated (r = 0.80, 95% confidence interval 0.47-0.93, p < 0.001). Urinary excretion of hypoxanthine increased almost 40 times from 0.12 +/- 0.14 to 4.6 +/- 2.9 mumol/kg/h indicating that renal excretion of hypoxanthine is not achieved just by passive filtration. We conclude that in newborn piglets hypoxanthine can pass from plasma to CSF and vitreous humor. Thus an increased CSF hypoxanthine concentration is not definite proof that significant cerebral hypoxia has occurred. PMID- 8229613 TI - Steroid usage in pregnancies complicated by preterm premature rupture of the membranes. AB - The aim of this study was to determine if increased antenatal steroid usage might account for the apparent protective effective of preterm premature rupture of the membranes (PPROM) against neonatal respiratory distress. Comparison was made of the outcome of 36 preterm infants consecutively admitted to the neonatal unit whose mothers had PPROM (study patients) and 36 matched controls whose mothers had intact membranes. The control infants were matched for gestational age at delivery, gender, mode of delivery and approximate date of birth. We found that significantly more study patients had received antenatal steroid therapy, 14 or 36 compared to five of 36 controls, p < 0.03. The study group tended to require a shorter duration of mechanical ventilation and a lower maximum inspired oxygen concentration. However, when matched infants were compared, neither of whose mothers had received antenatal steroids, this trend was reversed with the study patients requiring more respiratory support. We conclude these results suggest that increased antenatal steroid usage may account for the apparent protective effect of PPROM against the development of neonatal respiratory distress. PMID- 8229614 TI - Erythropoietin concentrations in amniotic fluid and umbilical venous blood from Rh-immunized pregnancies. AB - We set out to investigate prospectively the levels of erythropoietin in amniotic fluid and umbilical venous blood, and to attempt to relate these to fetal haemoglobin and lactate concentrations and to pCO2 and PO2 in Rh immunised patients studied before the onset of labor. Fetal blood was obtained by cordocentesis, and amniotic fluid by amniocentesis from a consecutive series of 36 Rh immunized patients at the time of fetal blood sampling. There was a close correlation (tau = 0.357, P = 0.0001) between the concentrations of erythropoietin in umbilical venous blood and those in amniotic fluid. Erythropoietin in umbilical venous blood correlated inversely with hemoglobin (tau = 0.453, P = 0.0001), and directly with lactate concentrations (tau = 0.450, P = 0.0005). When all other variables were considered, multiple regression analysis demonstrated hemoglobin concentration to be the only variable to be related to the level of erythropoietin in umbilical venous blood taken before transfusion. When the same analysis was performed on the same variables, adding erythropoietin concentration in amniotic fluid as the dependent variable, only erythropoietin in umbilical venous blood was found to be related to the level of erythropoietin in amniotic fluid. We conclude that the erythropoietin concentration in umbilical venous blood from Rh-immunized patients before the onset of labor, is related to fetal anemia. We also conclude that erythropoietin concentration in amniotic fluid is related to that in fetal blood, thereby indicating that the fetus is an important source of amniotic fluid erythropoietin in non laboring patients. PMID- 8229615 TI - Low dose aspirin in pregnancy: a clinical and biochemical study of effects on the newborn. AB - Low dose Aspirin in pregnancy reduces the incidence of intra uterine growth retardation (IUGR) and pregnancy induced hypertension (PIH) in women at risk for these complications. To investigate if this drug, even in a low dose, could expose the newborn to hemorrhagic complications, we studied ten neonates whose mothers had been taking 50 mg/day of Aspirin from the 12th week of pregnancy until delivery and compared them with eight newborns whose mothers didn't take the drug. No hemorrhagic complications (emathemesis, ecchymoses or petechiae, subconjunctival hemorrhage, cephaloematomas etc.) were observed in the fetuses exposed to Aspirin or in the control group. No hemorrhagic lesions were found by ultrasound brain scan on the fourth day of life. Newborns exposed to Aspirin showed a significantly lower thromboxane concentration on the first day of life (median 73 ng/ml versus 217 ng/ml); however on the fourth day the level of serum thromboxane in the cases exposed reached the values of the unexposed ones (median 146 ng/ml versus 143 ng/ml). In conclusion low dose Aspirin in pregnancy can be considered a safe drug without and adverse effect on the newborn. PMID- 8229616 TI - Prenatal ultrasonographic diagnosis of acardiac twin. AB - Acardiac twinning is a rare complication of monozygotic twin gestation occurring in 1/35,000 pregnancies or in 1% of all monozygotic twins. This condition is characterized by partial or complete lack of development of the heart in one of the twins, and requires that the normal twin (pump twin) provides circulation for itself as well as the acardiac sibling (perfused twin) by means of reverse circulation through large artery-to-artery and vein-to-vein anastomoses. The acardiac anomaly is uniformly fatal for the perfused twin, and the perinatal mortality for the normal twin is about 50%, usually as the result of congestive heart failure, polyhydramnios, and preterm delivery. In this report we present two cases of acardiac twin prenatally diagnosed by ultrasound. The principal sonographic features for prenatal diagnosis and the clinical management are briefly discussed. PMID- 8229617 TI - Pregnancy complicated by the May-Hegglin anomaly. AB - May-Hegglin anomaly is a prepotent autosomal hereditary disease characterized by giant thrombocytes, thrombocytopenia and continuous appearance of inclusion bodies (Dohle like corpuscle) in the cytoplasm of granulocytes. In this report we report our experience of a case whose pregnancy was complicated by this disease. This anomaly is rare and there are only 15 families reported in this country. The only clinical symptom is the tendency to bleed, but since this is not rare, it may be necessary to consider examination of peripheral blood in every case of thrombocytopenia. If this disease complicates pregnancy, it is important to plan the mode of delivery to consider both the maternal and fetal prognosis. PMID- 8229618 TI - A case of occipital encephalocele combined with right lung aplasia in a twin pregnancy. AB - A case of occipital encephalocele combined with right-sided pulmonary agenesis in one sibling of a twin gestation is presented. Both, encephalocele and lung aplasia, are rare conditions, the former occurring with a frequency of 1:3000 1:30,000, the latter in 1:10,000-1:30,000. Although both conditions may show concomitant malformations, to our knowledge this is the first case of a combination of these two malformations to be reported. PMID- 8229619 TI - Small versus large particles of demineralized freeze-dried bone allografts in human intrabony periodontal defects. AB - Various particle sizes of demineralized freeze-dried bone allograft (DFDBA) are currently used to treat patients with periodontal osseous defects. However, the effect of particle size on the healing of human intrabony periodontal defects is unknown since there have been no direct clinical comparisons. The purpose of this study was to compare the bony defect resolution obtained using two different particle size ranges of DFDBA. Cortical bone from a single donor was processed and ground to final particle sizes of 250 mu to 500 mu or 850 mu to 1,000 mu using an analytic mill. Paired interproximal intrabony periodontal defects in 11 patients were grafted with DFDBA. Soft and hard tissue measurements were made using an electronic constant-force probe at the initial and reentry surgeries. Treated sites in 10 patients were reevaluated by reentry approximately 6 months postoperatively. Mean bony defect fill was 1.66 mm for the large particle group and 1.32 mm for the small particle group. There was no statistically significant difference in bony fill between defects grafted with the different particle sizes of DFDBA when used in humans. PMID- 8229620 TI - The effects of incubation length and temperature on the specificity and sensitivity of the BANA (N-benzoyl-DL-arginine-naphthylamide) test. AB - A previous multi-center study examining patients diagnosed as having at least four periodontally diseased teeth showed that when BANA (N-Benzoyl-DL-Arginine Naphthylamide) hydrolysis by periodontal pathogens such as Treponema denticola, Porphyromonas gingivalis, and Bacteroides forsythus was evaluated versus clinical parameters such as clinical judgment of disease, bleeding on probing, and pocket depth, the sensitivity of the test was 84%, 82%, and 87%, respectively, while the specificity was only 42%, 41%, and 32%, respectively. The purpose of the present investigation was to improve the specificity of the test while retaining a high level of sensitivity in both gingivally healthy and periodontally diseased groups. One hundred forty-nine patients participated in this study providing 3,497 interproximal plaque samples. Gingival health was measured using the papillary bleeding score and this was compared with the presence or absence of detectable trypsin-like activity, as determined by the BANA hydrolysis of interproximal plaque samples, using a commercially-available test. Sensitivity and specificity were measured by varying the incubation time and temperature of the enzymatic assay. Using the correlated binomial model to analyze site-specific data within a patient, the specificity was highest at 35 degrees C and 5 minutes incubation (94%), and lowest at 45 degrees C and 15 minutes incubation (33%). Sensitivity was highest at 45 degrees C or 55 degrees C and 15 minutes incubation (90%) and lowest at 35 degrees C and 5 minutes incubation (47%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229621 TI - Prevalence of periodontal disease in a health maintenance organization and comparisons to the national survey of oral health. AB - The purposes of this study were to: 1) characterize the demographics, oral health behavior, and periodontal status of a health maintenance organization sample; 2) investigate the relationship between the location of posterior proximal measurement sites and prevalence estimates for periodontal disease; and 3) compare the prevalence of persons with pockets > or = 4 mm in the present sample to the 1985 NIDR Survey of Oral Health. The sample consisted of 1,090 adults attending a large health maintenance organization. All proximal sites in one randomly selected posterior dental sextant were examined for probing depth using a constant force probe. Demographic, medical, and behavioral factors were determined by questionnaire. Results indicated that the sample consisted primarily of medically and periodontally-healthy middle-aged adults with good oral hygiene habits. Overall, the mean probing depth was 2.95 mm with 10.1% of sites/subject > or = 4 mm. A larger percent of subjects had probing depths with pockets > or = 4 mm at lingual proximal sites than buccal proximal sites. Prevalence of subjects with pockets > or = 4 mm at mesio-buccal sites in the present study was similar to NIDR Region III data (15.3% vs. 17.4% respectively). However, when data from all posterior sites were included, the overall prevalence rate in the present sample increased to 36.8%. These findings indicate that disease prevalence is dependent on the location of surfaces measured and conservatively indicate that NIDR survey data may have underestimated the prevalence of persons with periodontal pockets > or = 4 mm by at least 20%. PMID- 8229622 TI - Evaluation of a counter-rotational powered brush in patients in supportive periodontal therapy. AB - In order to evaluate the effectiveness of a counter-rotational powered brush (CRPB) during the supportive periodontal therapy (SPT) phase of periodontal treatment, 40 treated patients in SPT but with insufficient plaque control were randomly divided into equal experimental or control groups. All subjects used the same toothpaste, but the CRPB group did not use any interproximal aids. Gingivitis (MGI), plaque, and bleeding on probing (BOP) were scored at baseline and 1, 3, and 6 months prior to prophylaxis in conjunction with regular SPT visits. While both groups improved from baseline, CRPB use achieved significantly lower mean plaque scores and BOP at 6 months as analyzed by repeated measures ANOVA. The CRPB also showed consistent statistically superior percentage changes from baseline resulting in a general 50% improvement in clinical conditions compared to a 20 to 25% improvement for control oral hygiene methods. CRPB use resulted in at least a 50% improvement from baseline twice as often as did the control. The results of this study demonstrate more substantial and consistent improvement in periodontal conditions and plaque control effectiveness with the CRPB than the control methods that included interproximal hygiene aids. It appears that the CRPB may be a useful adjunct in maintaining reduced plaque levels and favorable gingival conditions in patients in the SPT phase of periodontal therapy. PMID- 8229623 TI - A cross-sectional study on osteocalcin levels in gingival crevicular fluid from periodontal patients. AB - The purpose of the present study was to determine the levels of osteocalcin, a bone specific matrix protein, in gingival crevicular fluid (GCF) from periodontal disease patients and to investigate the relationship between GCF osteocalcin levels and clinical parameters. Nineteen initial visit patients, 5 patients with gingivitis and 14 patients with adult periodontitis, participated in this study. The clinical parameters including probing depth, attachment level, gingival index, and tooth mobility were recorded following careful sampling of GCF with a filter paper strip harvested for 3 minutes. Osteocalcin adsorbed on a strip was extracted in a plastic tube containing 150 microliters of 10 mM sodium phosphate buffer (pH 6.5). GCF osteocalcin was determined by a newly-developed, high sensitive enzyme immunoassay which could recognize the N-terminal 20 residue peptide. In gingivitis patients, no significant amounts of osteocalcin were detected. In periodontitis patients, on the other hand, osteocalcin levels were detected, ranging between 0 and 540 pg/tube and positively correlated with these clinical parameters (P < 0.01). Moreover, in several sites in GI = 3 group, extremely higher levels of GCF osteocalcin were detected. These results strongly suggest that in addition to the presence of GCF osteocalcin the levels of osteocalcin may reflect the degree of the periodontal inflammation at the sampled sites. PMID- 8229624 TI - Effect of a collagen matrix on healing in periodontal fenestration defects in dogs. AB - Contralateral periodontal fenestration defects in seven beagle dogs were used to evaluate influence of a collagen matrix on periodontal wound healing. The defects (6 x 4 mm) were created through the buccal cortical plates of the maxillary canine teeth following elevation of mucoperiosteal flaps. The collagen was fitted to the defects on one side. Contralateral defects served as controls. Flaps were repositioned and sutured. Dogs were sacrificed 4 weeks after surgery and block sections including teeth and surrounding structures were prepared for histometric analysis. No meaningful differences in cementum and bone regeneration were observed between treatments. There was seemingly more bone regeneration in the apical than in the coronal aspect of the defects and significantly more cementum regeneration. Root resorption was observed in one collagen and one control defect. Ankylosis was not observed. The results suggest that the maxillary canine periodontal fenestration defect can be used as a model to evaluate factors that may enhance cementum and bone regeneration. The collagen matrix neither enhanced nor inhibited periodontal wound healing in this model. PMID- 8229625 TI - Periodontal repair in dogs: expanded polytetrafluoroethylene barrier membranes support wound stabilization and enhance bone regeneration. AB - A wound stabilizing effect of expanded polytetrafluoroethylene (ePTFE) membranes was evaluated in supra-alveolar periodontal defects in 5 beagle dogs. The defects, 5 to 6 mm in height, were surgically created around the 2nd, 3rd, and 4th mandibular premolar teeth in contralateral jaw quadrants. The root surfaces were conditioned with heparin, which, in this model, has been demonstrated to compromise periodontal healing and result in formation of a long junctional epithelium. Wound closure included application of ePTFE membranes around each premolar tooth in one jaw quadrant in each dog and flap positioning coronal to the cemento-enamel junction in both jaw quadrants. Healing progressed uneventfully except for 3 teeth in 2 dogs, which experienced membrane exposure. The dogs were sacrificed after a 4-week healing period and tissue blocks were prepared for histometric analysis. Connective tissue repair in heparin+membrane treated teeth averaged 98% of the defect height compared to 84% in control heparin-treated teeth (P < or = 0.05). Junctional epithelium formation was smaller in membrane-treated teeth than in control teeth (P < or = 0.05) and was usually terminated coronal to the membrane. Bone regeneration was enhanced in membrane-treated teeth compared to controls (P < or = 0.01) and was strongly correlated to the area under the membrane in teeth without membrane exposure (r2 = 0.993; P = 0.002). This correlation was reduced when teeth with membrane exposure were included in the analysis (P < or = 0.05). Cementum regeneration was minimal under both treatment conditions. Root resorption was increased in membrane-treated compared to control teeth (P < or = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229626 TI - Six-year progression of destructive periodontal disease in 2 subgroups of elderly Chinese. AB - Two groups of elderly chinese were selected from a large epidemiological sample on the basis of a low ("best" group) or a high ("worst" group) number of sites with attachment loss levels > or = 6 mm and/or pocket depth > or = 4 mm and at least 16 teeth present. Six years later the patients were clinically reexamined and the subgingival microflora was assessed. This paper presents the clinical characteristics of destructive periodontal disease progression among the two subgroups. The "best" group lost an average of 1.8 teeth, contrasting the average loss of 5.3 teeth among the "worst" group. Virtually all teeth lost among the "worst" group had a baseline attachment loss level > or = 4 mm, in contrast to 48% among the "best" group. While dental caries could be identified as a cause of tooth loss in both groups, the excess tooth loss among the "worst" group seems attributable to periodontal destruction. The average of 1.21 mm attachment/site lost among the "best" group was not statistically significantly different from the 1.36 mm/site lost among the "worst" group during the 6 years. Individual mean losses of attachment ranged from a gain of 0.03 mm to a loss of 3.19 mm. An attachment loss > or = 2 mm at a site was highly positively associated with a high initial attachment loss level (> or = 4 mm) at that site among the "best" group, whereas a highly negative association was seen among the "worst" group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229627 TI - Gingival recession in relation to history of hard toothbrush use. AB - Gingival recession studies in the U.S. have related primarily to sex and age with little consideration of toothbrush hardness. This preliminary study examined the relation between a history of hard toothbrush use and gingival recession. A total of 182 subjects, male and female, between 18 and 65 years of age, with a minimum of 18 natural teeth, no advanced periodontitis or history of periodontal surgery were examined. Gingival recession was scored as present whenever the free gingival margin was apical to the cemento-enamel junction and root surface was exposed. History of hard toothbrush use was ascertained. Eighty-two subjects had a history of hard toothbrush use, 77 did not, and 23 did not know. The percentage of subjects with recession increased with age from 43% to 81%, with a figure of 63% for all age groups combined. Males tended to show slightly greater levels of recession than females. Regression analysis showed that females had about 4 percentage points less receded surfaces than males. Recession was also found to be more pronounced for subjects with a history of hard toothbrush use, with a mean of 9.4% receded surfaces versus 4.7% for those who had never used a hard brush. For users of hard toothbrushes, the percent of surfaces with recession showed a significant and dramatic increase with increasing brushing frequency; this effect did not exist for those without a history of hard brush use. The relation with age was highly significant, with regression analysis showing that the percent of surfaces with recession tends to increase about 3.5 percentage points per decade.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229628 TI - Microscopic and chemical analysis of bone-hydroxyapatite interface in a human retrieved implant. A case report. AB - The aim of the present study was a histological and chemical analysis of the bone hydroxyapatite interface in a retrieved human implant. The implant was cut in two: one half was processed for light microscopy with the cutting grinding system, and the other for SEM analysis with EDX and a cathodoluminescence system. Bone maintained an intimate contact with the hydroxyapatite and the bonding of the hydroxyapatite with the metal was very strong. An analysis of the interface from a double staining (basic fuchsin and von Kossa) showed a non-mineralized matrix, interposed between hydroxyapatite and bone in many areas. It is possible that different microscopic features are present in different areas of the interface, however, intimate contact was observed 6 months after insertion of the implant. PMID- 8229629 TI - Erythema multiforme involving gingiva. AB - Erythema multiforme is a vesiculobullous condition that may affect skin and/or mucosa. Oral lesions are characterized by hemorrhagic crusting of the lips and ulceration mainly of the non-keratinized mucosa. This paper describes a patient who presented with gingival lesions as well as the more typical oral signs of erythema multiforme. PMID- 8229630 TI - Re: Localized osteomyelitis secondary to endodontic implant pathosis (J Periodontol; 1993;64:306-310) PMID- 8229631 TI - Re: Periodontal terminology. PMID- 8229632 TI - Re: Treatment of periodontal diseases with metronidazole (J Periodontol 1993;64:1 15) PMID- 8229633 TI - Re: Biologic width and crown lengthening,. PMID- 8229634 TI - The meaning of low scores on MMPI-2 clinical scales of normal subjects. AB - Low scores often have been ignored in validity research on the MMPI, yet these statistically significant deviations from the norm may be valuable sources of information and occur frequently enough to merit further investigation. The meaning of low scores on the MMPI-2 was examined for 822 male-female partner pairs in the MMPI-2 restandardization sample. Subjects were divided by MMPI-2 scale scores into high-, medium-, and low-score groups. Analyses of variance were performed with score level as the independent variable and partner ratings as dependent variables. Significant differences were found between the low- and medium-score groups with low scorers rated as better adjusted that medium scorers. More significant differences were found between the high-score and medium-score groups than between the low-score and medium-score groups; this suggested that high scores on MMPI-2 clinical scales are associated with poorer adjustment. PMID- 8229635 TI - Personality variables in cocaine- and marijuana-dependent patients. AB - Khantzian (1985) hypothesized that the drug of choice of patients with substance dependence disorders reflects an attempt at self-medication. Cocaine- and marijuana-dependent inpatients were compared using the MMPI to test this hypothesis. These two groups of patients did not differ on the standard validity and clinical scales of the MMPI, and their scores were basically similar to a group of alcohol-dependent inpatients and a group of psychiatric patients of similar age. There was no single MMPI code type that was characteristic of either group of substance-dependent patients. It appears that drug of choice had little effect on the MMPI scores of these two groups of substance-dependent patients. These results are not consistent with the theory of self-medication. PMID- 8229636 TI - The relationship between MCMI personality scales and clinician-generated DSM-III R personality disorder diagnoses. AB - This study examined the relationship between elevations on the personality scales of the Millon Clinical Multiaxial Inventory (MCMI) and clinician-generated Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev. [DSM-III-R]; American Psychiatric Association, 1987) diagnoses for 101 psychiatric patients at a VA medical center/psychiatric hospital in the Southeastern United States. Personality disorder diagnoses were made by employing a personality symptom checklist that consisted of all the verbatim criteria for personality disorders contained in the DSM-III-R. Clinicians who completed the checklists were required to have had at least 5 hr of direct contact with the patients who completed the MCMI. The results indicated that only the Schiozotypal scale of the MCMI was related to its respective DSM-III-R personality disorder in the simple correlation. An examination of the diagnostic efficiency statistics for each of the MCMI personality disorder scales revealed overall low sensitivity, poor specificity, poor positive predictive power, and low diagnostic power, which suggests that the MCMI may have only limited utility in identifying personality disorders. PMID- 8229637 TI - Hand test characteristics of dual diagnosed mentally retarded older adults. AB - The projective Hand Test (Wagner, 1962/1983) was administered to 17 older adults with a dual diagnosis (mental retardation existing concurrently with some form of mental illness) residing in a Midwestern institution for the mentally retarded. Each of these individuals was matched with another resident in the same institution with a single diagnosis of mental retardation on the variables of age, sex, and level of mental retardation to determine differences between the groups on the Hand Test. The dual diagnosis group gave significantly more bizarre (BIZ) responses, whereas the single diagnosis group gave more descriptive (DES) responses. It was contended that these two variables accurately reflect the essential difference between dual and single diagnosed mentally retarded individuals. PMID- 8229638 TI - Psychodiagnosis of personality structure. II: Borderline personality organization. AB - The borderline conditions have been the focus of considerable debate and controversy for over 30 years. This article, second in a series of three articles focusing on Kernberg's (1975, 1984) psychostructural diagnosis of personality organization, examines Rorschach contributions to the description and diagnosis of borderline personality organization. Rorschach approaches integrating nomothetic and idiographic data are applied to borderline personality functioning and appraised in light of the Rorschach's contribution to the clarification of the controversial and poorly delineated borderline diagnosis. PMID- 8229639 TI - A borderline psychopath: "I was basically maladjusted...". AB - The case of a 21-year-old White man who committed a robbery and homicide is presented. His Rorschach is analyzed and interpreted using both psychostructural (Exner, 1986a) and psychodynamic (Cooper, Perry, & Arnow, 1988; Gacono & Meloy, 1992; Kwawer, 1980) methodologies. Findings are used to understand the presence and interaction of both psychopathic character and borderline personality organization in this explosive and sadistic young man. PMID- 8229640 TI - Melatonin and sleep in humans. AB - Early studies on the physiological effects of melatonin typically reported hypnotic 'side-effects'. Later studies, specifically addressing this action, failed to reliably replicate hypnotic effects using standard polysomnography. This difference may be related to differences in the basic physiological action of melatonin compared with more conventional hypnotics. It is suggested that melatonin exerts a hypnotic effect through thermoregulatory mechanisms. By lowering core body temperature, melatonin reduces arousal and increases sleep propensity. Thus, in humans, one role of melatonin is to transduce the light-dark cycle and define a window-of-opportunity in which sleep-propensity is enhanced. As such, melatonin is likely to be an effective hypnotic agent for sleep disruption associated with elevated temperature due to low circulating melatonin levels. The combined circadian and hypnotic effects of melatonin suggest a synergistic action in the treatment of sleep disorders related to the inappropriate timing of sleep and wakefulness. Adjuvant melatonin may also improve sleep disruption caused by drugs known to alter normal melatonin production (e.g., beta-blockers and benzodiazepines). If melatonin is to be developed as a successful clinical treatment, differences between the pharmacological profile following exogenous administration and the normal endogenous rhythm should be minimized. Continued development as a useful clinical tool requires control of both the amplitude and duration of the exogenous melatonin pulse. There is a need to develop novel drug delivery systems that can reliably produce a square-wave pulse of melatonin at physiological levels for 8 10 hr duration. PMID- 8229641 TI - Immunohistochemical study of the pineal astrocytes in the postnatal development of the cat and dog pineal gland. AB - The expression of glial antigens vimentin (VIM) and glial fibrillary acidic protein (GFAP) is described in the pineal gland of cats and dogs from the first postnatal days to adulthood. VIM immunopositive cells were observed from the first postnatal days in both species. GFAP expression starts from the second postnatal week. In adults, a notable population of stellate cells immunopositive for GFAP and VIM was found dispersed throughout the gland. According to their immunocytochemical profile, these cells could be identified as astrocytes. PMID- 8229642 TI - One-hour exposure to moderate illuminance (500 lux) shifts the human melatonin rhythm. AB - Salivary melatonin levels were measured in 12 healthy volunteers in order to determine whether a moderate light intensity, which suppresses the nocturnal rise of melatonin, was able to shift the melatonin rhythm. The samples were collected at 1-hr intervals under lighting of < 100 lux (experiment 1) or < 10 lux (experiment 2). The control melatonin profiles were determined during the first night. In the second night the subjects were exposed to light of 500 lux for 60 min during the rising phase of melatonin synthesis. The third series of samples was collected during the third night. The mean decrease of melatonin levels by the exposure to light was 56% of the prelight concentrations. The melatonin onset times were delayed significantly (about 30 min) the night after the exposure to light. The melatonin offset times tended to be delayed in experiment 2. The shifts of the melatonin offset correlated positively with the amount of the melatonin suppression. The results suggest that a relatively small and short lasting light-induced interruption of melatonin synthesis may affect the melatonin rhythm in humans. PMID- 8229643 TI - Effect of pinealectomy and plane of nutrition on wool growth in Merino sheep. AB - The effects of surgical pinealectomy and plane of nutrition on wool growth and plasma prolactin concentrations in young Merino wethers were investigated. In young pinealectomized wethers maintained at a low live weight under conditions of minimal stress, the decline of conditioned clean wool production on midside patches was slowed when compared to pineal-intact controls. This difference appeared to be due in part to the observed greater secondary wool follicle density in the pinealectomized wethers; mean fiber diameter was affected to a smaller extent, while staple length growth rate was not significantly altered. Circulating prolactin profiles showed a seasonal variation (high in summer, low in winter) in both pinealectomized and control wethers. There was no difference in wool production between pinealectomized and control wethers when the diet of the same wethers was subsequently supplemented with formaldehyde-treated cottonseed meal. While the role of melatonin in the regulation of wool growth remains to be determined, it is suggested that the hormone may have a transient effect on wool production in young wethers under conditions of limited nutrition, but not at higher nutritional levels, and that melatonin may be involved in partitioning of nutrients to the wool follicle. PMID- 8229644 TI - Hydroxyindole-O-methyltransferase activity in ocular and brain structures of rabbit and hen. AB - Relative activities of hydroxyindole-O-methyltransferase (HIOMT) of some brain and ocular structures of the rabbit and hen were analyzed using different 5 hydroxyindoles, i.e., N-acetylserotonin (NAS), 5-hydroxytryptophol (HTOL), 5 hydroxytryptophan (HTP), 5-hydroxytryptamine (HT), and 5-hydroxy-3-indoleacetic acid (HIAA), as enzyme substrates. Pineal glands of both species, as well as hen retina, are capable of producing, to varying degrees, melatonin, 5 methoxytryptophol, and 5-methoxytryptamine. Hen choroid and iris-ciliary body O methylated NAS and HTOL, whereas rabbit choroid and, to a much lesser extent, hypothalamus and cerebral cortex all O-methylated only NAS. No measurable HIOMT activity was found in hen brain. NAS was a preferred substrate for HIOMT in the hen tissues, whereas in the rabbit pineal gland NAS and HTOL were equally good substrates for HIOMT. Other tested 5-hydroxyindoles, i.e., HTP, HT, and HIAA, were poor methyl acceptors. Of the tissues examined, the highest HIOMT activity was found in the hen pineal gland, followed by the rabbit pineal gland and hen retina. No significant differences between day and nighttime enzyme activities were observed in the pineal gland and retina of either species. The data suggest that in vertebrates some nervous and ocular tissues possess the potential to produce 5-methoxyindole compounds; however, the HIOMT-catalyzed process shows remarkable substrate-, tissue- and species-dependent variations. PMID- 8229645 TI - Vasopressin and vasoactive intestinal peptide infused in the paraventricular nucleus of the hypothalamus elevate plasma melatonin levels. AB - The connection between the suprachiasmatic nucleus (SCN) and the paraventricular nucleus of the hypothalamus (PVN) forms an important component of the melatonin rhythm-generating system. However, the chemical identity of this projection is not known. To test the possible implication of the SCN peptides vasopressin (VP) and vasoactive intestinal peptide (VIP) in this projection, we performed microinfusions in the PVN during the first half of the dark period and subsequently monitored resulting plasma melatonin levels. Infusions for 7 hr of either VP or VIP, but not oxytocin, caused increased plasma melatonin levels in the middle of the dark period. These observations confirm the role of the PVN in the melatonin rhythm-generating pathway and indicate that both VP and VIP released at the level of the PVN, and probably derived from the SCN, are able to influence peripheral plasma melatonin levels. PMID- 8229646 TI - An immunohistochemical study of neuropeptide Y in the bovine pineal gland. AB - An immunohistochemical study of the bovine pineal gland was performed using rabbit polyclonal antibodies raised against neuropeptide Y (NPY) or against the C terminal flanking peptide of proNPY (CPON). A large number of NPY/CPON immunoreactive (IR) nerve fibers were demonstrated throughout bovine pineal gland. The IR-fibers were located in the capsule of the gland, usually piercing into the gland together with blood vessels. In the gland itself, the fibers were also located intraparenchymally between the pinealocytes. Within the rostral and caudal areas of the pineal stalk, NPY-IR fibers were also observed, and these fibers could be followed not only into the gland but also to the habenular and posterior commissures. The morphological localization of the NPY-IR nerve fibers in the bovine pineal gland indicate that the majority of fibers originate from the superior cervical ganglion. However, some fibers probably originate from the brain itself. PMID- 8229647 TI - Selective inhibition of NS-398 on prostanoid production in inflamed tissue in rat carrageenan-air-pouch inflammation. AB - NS-398 (N-(2-cyclohexyloxy-4-nitrophenyl) methane sulphonamide), a newly synthesized potent non-steroidal anti-inflammatory drug (NSAID) has a much lesser degree of toxicity, as compared with presently available NSAIDs. We have investigated the inhibition of prostanoid production in inflammatory exudate, gastric mucosa and renal papillary tissue, following oral administration to carrageenan-air-pouch rats. The ID50 values of NS-398 in the inflammatory exudate, gastric mucosa and renal papillary tissue were 0.18, 62.2 and 261.7 mg kg-1, respectively. In contrast, indomethacin decreased the PGE2 concentration in the inflammatory exudate, gastric mucosa and renal papillary tissue, with the same dose range, the ID50 values being 0.23, 0.14 and 0.15 mg kg-1, respectively. The same tendency was seen for 6-keto-prostaglandin F1 and thromboxane B2. Moreover, NS-398 inhibited excess PGE2 production in inflamed tissue but did not affect physiological production of PGE2 in non-inflamed tissue. Indomethacin, in both inflamed and non-inflamed tissues, inhibited PGE2 production to the same degree. These results indicated that NS-398 has some specificity for inflamed tissue, by inhibiting prostanoid synthesis, and this effect may explain the decreased side-effects of this drug. PMID- 8229648 TI - Affect, culture, and morality, or is it wrong to eat your dog? AB - Are disgusting or disrespectful actions judged to be moral violations, even when they are harmless? Stories about victimless yet offensive actions (such as cleaning one's toilet with a flag) were presented to Brazilian and U.S. adults and children of high and low socioeconomic status (N = 360). Results show that college students at elite universities judged these stories to be matters of social convention or of personal preference. Most other Ss, especially in Brazil, took a moralizing stance toward these actions. For these latter Ss, moral judgments were better predicted by affective reactions than by appraisals of harmfulness. Results support the claims of cultural psychology (R.A. Shweder, 1991a) and suggest that cultural norms and culturally shaped emotions have a substantial impact on the domain of morality and the process of moral judgment. Suggestions are made for building cross-culturally valid models of moral judgment. PMID- 8229649 TI - "Who am I? Why am I here?": self-esteem, self-clarity, and prototype matching. AB - Recent research has demonstrated that individuals with low self-esteem lack self clarity; they have less certain and less stable self-concepts than do those with high self-esteem (A. H. Baumgardner, 1990; J. Campbell, 1990). Self-concept confusion should mitigate against the use of a decision-making strategy that involves using the self to guide choice behavior (i.e., prototype matching, P. M. Niedenthal, N. Cantor, & J. F. Kihlstrom, 1985). Two correlational studies demonstrated that people with high self-esteem, but not low self-esteem, made use of prototype matching in forming preferences. In a 3rd study, the self-concept was made more clear or made more confused. Clarity was associated with the use of prototype matching regardless of level of self-esteem. Self-concept confusion was associated with a failure to use the strategy regardless of level of self-esteem. PMID- 8229650 TI - What motivates children's behavior and emotion? Joint effects of perceived control and autonomy in the academic domain. AB - This study examined the contribution of perceived control and autonomy to children's self-reported behavior and emotion in the classroom (N = 246 children ages 8-10 years). Multiple regression analyses revealed unique effects of autonomy over and above the strong effects of perceived control. In addition, both sets of perceptions (and their interaction) were found to distinguish children who were active but emotionally disaffected from those who were active and emotionally positive. Specific predictions were also tested regarding the effects of (a) control attributions to 5 causes and (b) 4 reasons for task involvement that differed in degree of autonomy on children's active (vs. passive) behavior and 4 kinds of emotions: boredom, distress, anger, and positive emotions. Implications of the findings for theories of children's motivation are discussed, as well as for diagnostic strategies to identify children at risk for motivational problems PMID- 8229651 TI - Ego development and individual differences in personality. AB - The relation between individual differences in personality and differences in developmental maturity was studied by relating observations of personality by multiple, independent judges to level of ego development. The personality characteristics of longitudinally followed Ss (104 at age 14; 98 at age 23) were evaluated by the California Adult Q-Set (CAQ); ego level was evaluated by the Washington University Sentence Completion Test of Ego Development (SCT). A priori personality dimensions--consisting of CAQ items for which a common developmental pathway was expected--were constructed and related to the SCT: (a) Ego-resiliency and interpersonal integrity were associated with increasing ego development, (b) conformity was associated with the Conformist level and, unexpectedly, to the Conscientious level, (c) need regulation was associated with the Conscientious level, and (d) self-ease and expressiveness-playfulness were not associated with ego level. PMID- 8229652 TI - Religion's role in adjustment to a negative life event: coping with the loss of a child. AB - Parents (N = 124) who had lost an infant to sudden infant death syndrome were interviewed 3 weeks and 18 months postloss. Two components of religion (religious participation and religious importance) were assessed, and their relations with 3 coping-process variables (perceived social support, cognitive processing of the loss, and finding meaning in the death) were examined. Greater religious participation was related to increased perception of social support and greater meaning found in the loss. Importance of religion was positively related to cognitive processing and finding meaning in the death. Furthermore, through these coping-process variables, religious participation and importance were indirectly related to greater well-being and less distress among parents 18 months after their infants' deaths. Results suggest that further study of the social and cognitive aspects of religion would be profitable. PMID- 8229653 TI - Inhibition of murine retrovirus-induced immunodeficiency disease by dideoxycytidine and dideoxycytidine 5'-triphosphate. AB - The antiretroviral activity of many nucleoside analogues depends not only on their ability to inhibit the virus reverse transcriptase but also on the specific cellular pools of natural deoxynucleosides and on the level of the enzymes responsible for their phosphorylation. In an attempt to overcome these limitations, we have tested the efficacy of the oral administration of 2',3' dideoxycytidine (DDC) and the administration of its phosphorylated derivative, 2',3'-dideoxycytidine 5'-triphosphate (DDCTP) encapsulated into autologous red blood cells in a murine retrovirus-induced immunodeficiency model of AIDS (MAIDS). The results obtained showed that both single treatments are quite effective in preventing the typical signs of MAIDS. Combined treatment with both oral DDC and encapsulated DDCTP yields an additive response in some, but not all the parameters investigated. Furthermore, animals receiving the simultaneous administration of DDC and DDCTP show a reduction of animal body weight, a persistent high concentration of IgM, and a high titer of anti-LP-BM5 gag immunoglobulins. Thus, the administration of the same drug in different molecular forms and/or with different delivery systems should be carefully evaluated in preclinical animal models because of the unpredictability of the effects of these treatments from the conclusion drawn by studies on single treatment. PMID- 8229654 TI - Expansion of a CD8+CD28- cell population in the blood and lung of HIV-positive patients. AB - CD8+ cytotoxic T lymphocytes (CTLs) in human immunodeficiency virus (HIV) patients have defects in cytolytic ability and proliferative potential. The surface receptor CD28 is important in regulating antigen-specific responses to T cells. We hypothesized that activated CD8+ CTLs in HIV patients would have altered expression of CD28. We examined surface expression of D44, a CD8+ CTL marker, and of CD28 on T cells from blood and bronchoalveolar lavage from HIV+ patients and normal volunteers. We found no significant difference between normal volunteers and HIV+ individuals in percentage of CD8+D44+ CTLs in blood or the lung. In contrast, CD8+CD28- T cells in the blood of HIV patients constituted 74% of CD8+ cells compared to 25% in normal subjects (p = 0.001), findings exaggerated in both normal and HIV+ lung. CD4+CD28- blood T cells were significantly increased in HIV+ patients compared to normal subjects (24 vs. 1.5%, p = 0.004). The HIV infection itself did not directly downmodulate CD28 expression, demonstrated in the CD28+ SUPT1 cell line. Increased numbers of CD28- T cells may be the result of immunologic activation or of expansion of a preexisting CD28- subset. These findings have immunologic consequences for the antigen-specific response of T cells in HIV+ patients. PMID- 8229655 TI - Fusogenicity of mutant and chimeric proviruses derived from molecular clones of cytopathic and noncytopathic human immunodeficiency virus type 2. AB - Analysis of the phenotype of the molecular clones of cytopathic and fusogenic, noncytopathic and nonfusogenic, and chimeric proviruses of human immunodeficiency virus type 2 (HIV-2) suggests that the major determinant of the attenuated fusogenicity and cytopathicity of HIV-2 was located in the 3'-half of the genome, with envelope playing the more dominant role. However, no single linear domain within the envelope, including the major CD4 binding domain and fusogenic domain, was sufficient by itself for syncytia induction and cytopathic effects. Truncation of the transmembrane envelope glycoprotein downstream of the transmembrane region was not a major factor in this regard. However, truncation within the transmembrane region rendered the provirus replication incompetent. The regulatory genes (tat, rev) and auxiliary gene (nef) did not seem to play a critical role in determining HIV-2 fusogenicity in vitro. The results suggest the importance of the overall conformation of the envelope in the divergent phenotypes of HIV-2. PMID- 8229656 TI - Uric acid and didanosine compliance in AIDS clinical trials: an analysis of AIDS Clinical Trials Group protocols 116A and 116B/117. AB - An analysis is presented that promotes the use of uric acid levels as an indicator of patients didanosine (ddI) use. Logistic regression techniques were used on data from two test groups that best represent patients entered on ACTG protocols 116A and 116B/117. Two classification functions resulted that are based on serial uric acid measurements and were used to classify patients to one of two groups: those treated with zidovudine (ZDV) and those treated with ddI. These functions correctly classified well over 70% of the patients in each of these two studies. Implications of these results with respect to the assessment of ddI compliance and limitations in the use of these classification functions are discussed. PMID- 8229657 TI - Circadian rhythm of lymphocytes and their glucocorticoid receptors in HIV infected homosexual men. AB - Glucocorticoids (Gc) are known to modulate protein synthesis by immune cells through binding to a specific receptor (GcR). We outlined the circadian rhythm of plasma cortisol, ACTH, of the peripheral blood mononuclear cells (PBMC isolated by Ficoll-Hypaque technique), and of their subsets CD4, CD8 in 14 asymptomatic HIV+ homosexual men and in nine controls. We also estimated the GcR of the PBMC at 0700 and at 2300 hours, near the peak and nadir of the cortisol rhythm. In the HIV+ subjects, the PBMC circadian rhythm is abolished, an observation that confirms previous reports; in more than half of these patients, the GcR dissociation constant is larger than that of the controls. The circadian rhythms of plasma cortisol and ACTH levels do not differ from those of the controls. These changes may impair the function of the hypothalamic pituitary-adrenal axis in the HIV-infected subject. PMID- 8229658 TI - Geographic variation in human immunodeficiency virus seroconversion rates in the U.S. Navy. AB - The Navy population is centered around 19 U.S. home ports and several inland duty stations. This is a study of variation in human immunodeficiency virus (HIV) seroconversions in Navy enlisted men by location in the United States, based on 949,570 enzyme-linked immunosorbent assays and 812 seroconversions during 1987 90. Seroconversion rates were higher in personnel in San Francisco (p = 0.0004), Washington, D.C. (p = 0.001), and Orlando, FL (p = 0.04) than in other areas. They were lower in Charleston, SC, New London, CT, Seattle; and Brunswick, ME (p < 0.05). Black men had triple the seroconversion rate of all other men (p < 0.0001). After adjustment for race, age, marital status, and occupation, risk of seroconversion remained high in San Francisco (p = 0.02) and Orlando (p = 0.03). The risk of seroconversion in San Francisco was twice that of other areas in a cohort that did not change location (p = 0.01). In contrast to declining trends overall in the Navy, rates did not decrease in Washington, D.C., a result consistent with previous calculations indicating a delayed second wave of HIV infection in the region. PMID- 8229659 TI - Risk factors for HIV infection among injection drug users: results of blinded surveys in drug treatment centers, King County, Washington 1988-1991. AB - Among injection drug users (IDUs) entering drug treatment in King County, Washington between 1988 and 1991, we investigated HIV seroprevalence in relationship to demographic, sexual, and drug-use characteristics. Eighty-two of 3,039 (2.7%) IDUs tested HIV positive. Gay or bisexual men had the highest HIV prevalence (37.1%), followed by lesbian or bisexual women (8.3%), heterosexual men (2.3%), and heterosexual women (1.5%). American Indians were more likely to be infected with HIV than were whites. Those with no permanent address were more likely to be infected than those with an address. Unexpectedly, the prevalence of HIV infection among amphetamine injectors (13.1% of 168) was higher than among those who did not report using amphetamines. After adjustment for sexual orientation, HIV prevalence was four times higher among primary amphetamine injectors and three times higher among secondary amphetamine injectors than among injectors of other drugs. The basis for the strong association observed between HIV infection and a history of injection of amphetamines is not known and should be clarified through further research that obtains more detailed information on IDUs. PMID- 8229660 TI - HIV/AIDS seroincidence/seroprevalence studies from Africa. PMID- 8229661 TI - Virological diagnosis of mixed HIV-1/HIV-2 infection. PMID- 8229662 TI - [Research and development of new oral cephems, cefixime and cefdinir]. AB - This article reviews the structure-activity relationships, biological properties and synthesis of two new oral cephalosporin antibiotics, cefixime (CFIX) and cefdinir (CFDN). Our research into new oral cephalosporin antibiotics began in the late 1970's. The first goal of our research was to discover a new oral cephem possessing similar antibacterial activity and resistance to beta-lactamase as the 3rd generation injectable cephalosporins. We focused our attention on searching for a non-prodrug type cephem, that is, a new parent structure with high intrinsic absorption. We selected ceftizoxime (CZX) as a seed compound due to its relatively high excretion rate (8.5%) in the urine after oral administration to rats. We concentrated our research on the chemical modification of the oxime moiety in CZX based on our hypothesis for oral absorption, and discovered a lead compound with a carboxymethoxyimino group which displayed better urinary excretion (41.0%). Optimization studies led to a new oral cephem, CFIX. However, CFIX shows only low to moderate antibacterial activity against gram-positive bacteria such as S. aureus. Hence, the second goal of our research was to discover a new oral cephem with enhanced activity against gram-positive bacteria. From a consideration of structure-absorption relationships, we studied the activity and absorbability of cephems bearing other acidic functional groups at the oxime moiety of CFIX. As a result, we found a new oral cephem, CFDN with a hydroxyimino group at the 7-position. CFIX has excellent biological properties, displaying potent antibacterial activity against a wide range of gram-positive bacteria except S. aureus and gram-negative bacteria including opportunistic pathogens, high stability towards beta-lactamases and long acting efficacy. CFDN exhibits excellent and well balanced antibacterial activities against gram positive and gram-negative bacteria. The pharmacokinetic of CFDN in healthy volunteers showed that serum levels were high enough to make CFDN as an effective antibacterial agent. The proposed mechanisms of intestinal absorption of CFIX and CFDN are briefly described. The efficient synthetic methods to CFIX and CFDN were achieved via a common intermediate, 7-amino-3-vinylcephalosporanic acid diphenylmethyl ester from 7-ACA. PMID- 8229663 TI - [Study of iridaceae plants with the capability to induce differentiation of HL-60 promyelocytic leukemia cells]. AB - Thirteen species of Iridaceae plants were studied with the capability to induce HL-60 promyelocytic leukemia cells into macrophages. The neutrophil stimulating activity in these plants was also examined by measuring superoxide production. Both activities are known to be possessed by tumor promoters such as 12-O tetradecanoylphorbol-13-acetate (TPA). Dried parts of Iridaceae plants were extracted with methanol. Then, each extract was added to a culture medium of HL 60 cells and cultured for two days. A morphological change of the cells from suspended to adherent state, which was reported to occur in the process of differentiation, was observed. Nine species, most of which belong to the Iris genus, had the activity to cause cell adhesion. Each of the cell adhesion positive extract was applied to a silica gel column and eluted with chloroform, ethyl acetate and methanol in order. The activity was recovered only in the ethyl acetate eluate in all the cases. Those partially purified fractions also activated superoxide generation of neutrophils. When ethyl acetate fractions of Iris tectorum and Iris japonica were analyzed by HPLC, three active peaks different from TPA and teleocidins in UV spectra were detected. From these results, it was suggested that several plants of Iridaceae may contain TPA-type and yet structurally-different tumor promoters. PMID- 8229664 TI - [Synthesis and antiulcer effects of 5-hydroxy-4-[2-substituted-(E)-ethenyl]-2(5H) furanone. (1)]. AB - gamma-Hydroxybutenolides possessing conjugated substituents at the beta-position and their related compounds have been synthesized by the previously reported procedure with minor modifications and their antiulcer activities have been examined in the HCl-ethanol induced ulcer model often used for the evaluation of gastric mucosal protective factor enhancing effect. The compound A-1 5-hydroxy-4 [2-(2,6,6-trimethyl-1-cycohexenyl-1-yl)-(E)-ethenyl]-2 (5H)-furanone showed a pronounced effect at a low dosage of 5 mg/kg p.o. and some analogues compounds also exhibited potent inhibitory activity as compared with the reference drugs. The relationship between the structure of gamma-hydroxybutenolides and the antiulcer effect has been also examined and then the 5-hydroxyl group has been found to be essentially functional one to have antiulcer activity. PMID- 8229665 TI - [Inhibitory effect of bithionol on NADH-fumarate reductase in ascarides]. AB - To elucidate the mechanism of anthelmintic action of bithionol, the inhibitory effect of the drug on NADH-fumarate reductase (NADH-FR) of Ascaris lumbricoides suum was examined. NADH-FR, an enzyme of anaerobic carbohydrate metabolic pathway was solubilized from the mitochondria of the worm's muscle with deoxycholate, and then partially purified with the monoethanolamine-Sepharose 4B column chromatography. Rhodoquinone (RQ), which is required for the electron transfer from NADH to fumarate, was separated from the enzyme protein and phospholipids. Although the enzyme protein fraction eluted from the above column did not show NADH-FR activity, this enzyme was reactivated by the addition of purified RQ and phosphatidylcholine. The IC50 value of bithionol for reconstituted NADH-FR was 18 +/- 2 microM. The inhibition type was competitive to RQ. Bithionol inhibited at most 30% NADH-ferricyanide reductase, which did not require RQ, even at high concentration of 150 microM. These results suggest that the pharmacological action of bithionol, a phenolic anthelmintic, depends on the inhibition of the electron transport system by the competition with RQ. PMID- 8229666 TI - [Constituents of the leaves of Daphne pseudo-mezereum]. AB - Three new phenolic compounds were isolated from methanol extracts of the leaves of Daphne pseudo-mezereum (Thymelaeaceae) and characterized as (-)-pinoresinol di O-glucoside (1), 5-hydroxy-7-methoxy coumarin 8-O-beta-D-glucoside (2), diosmetin 7-O-beta-D-xylopyranosyl (1-->6) beta-D-glucopyranoside (3), respectively, together with six known compounds, using 1H- and 13C-NMR spectra. PMID- 8229667 TI - [Studies on pyridazinone derivatives. XV. Oxidative reactions and antibacterial antifungal activity of sulfur derivatives of pyridazinone]. AB - Oxidative reactions of o-nitrophenylthio substituted 2-phenyl-3(2H)-pyridazinones (3, 4, 13) with hydrogen peroxide were examined and o-nitrophenylsulfinyl compounds (5, 7, 14) and o-nitrophenylsulfonyl compound (6) were obtained. As the results of antibacterial antifungal screening tests in vitro, 3, 5 and 6 revealed strong antitrichophyton activities. PMID- 8229668 TI - Dodecylguanidine monoacetate (dodine) causes severe membrane damage in Pseudomonas syringae above the critical micelle concentration. AB - The release of K+ from Pseudomonas syringae cells treated with dodecylguanidine monoacetate (dodine) was followed with a K(+)-selective glass electrode. Treatment of the cells with 5-15 mumol/l dodine resulted in low levels of K+ release, but higher surfactant concentrations caused extensive and rapid K+ efflux. Dodine concentrations that caused high K+ release also induced significant leakage of inorganic phosphate. The addition of 5-10 mumol/l dodine also caused an increase in the rate of oxygen consumption in the presence of glycerol or succinate, but an increase in concentration from 10 to 40 mumol/l resulted in a concomitant decrease in O2 consumption. The results from this and previous work suggest that dodine inhibits respiration firstly by causing drainage of coenzymes, and then by a direct interaction with the components of the respiratory chain. Previous work showed that above 25 mumol/l, dodine molecules aggregate to form micelles. The results therefore suggests that, in contrast with other cationic amphiphiles, the micellar form of dodine is more damaging to the cytoplasmic membrane than the free molecules. PMID- 8229669 TI - Utilization of aromatic compounds by the Penicillium strain Bi 7/2. AB - The Penicillium strain Bi 7/2 utilized phenol, catechol, resorcinol, hydroquinone, pyrogallol, hydroxyhydroquinone, phloroglucinol, m- and p-cresol, orcinol, 4-methylcatechol, 4-methoxyphenol, 4-aminophenol, benzyl alcohol, benzoic acid, 2-, 3- and 4-hydroxybenzoic acid, anthranilic acid, protocatechuic acid and gallic acid as sole sources of carbon and energy. The central metabolites catechol, protocatechuic acid and hydroxyquinone could be determined by HPLC with diode-array detection. Pathways for the degradation of aromatic substances were proposed. PMID- 8229670 TI - Metabolism of PAH by fungi and correlation with extracellular enzymatic activities. AB - The activity to metabolize the polycyclic aromatic hydrocarbons (PAH) phenanthrene, anthracene, pyrene, fluorene and fluoranthene by Trametes versicolor, Pleurotus ostreatus (white rot fungi), Laetiporus sulphureus, Daedaela quercina, Flamulina velutipes (brown rot fungi), Marasmiellus sp. (litter decaying fungus) and Penicillium sp. M 1 (isolated from a PAH contaminated soil sample) were compared. Screening methods for the presence of exoenzymes (peroxidases, polyphenoloxidases, "radical generating" enzymes) were evaluated for their use in screenings for fungi degrading PAH. Laetiporus sulphureus and Penicillium sp. M 1 cometabolize several PAH with rates comparable to white rot fungi. In most of the cases the patterns of extracellular peroxidases indicate the potential of fungi to degrade PAH. PMID- 8229671 TI - Survey of mycoflora and mycotoxins of some dried fruits in Egypt. AB - Fifty-five species and two varieties appertaing to 23 genera were collected and identified from 4 samples of dried fig and 3 dried samples of each of apricot, plum and raisin. Forty-nine species and two varieties belonging to 20 genera were isolated on 1% glucose-Czapek's while 31 species and one variety belonging to 16 genera were isolated on 40% sucrose-Czapek's agar medium at 28 +/- 2 degrees C. Penicillium, Aspergillus and cladosporium were the common genera on the two types of media used. Alternaria and Pleospora were common on 1% glucose-Czapek's agar only while the genus of Eurotium was isolated with high occurrence on 40% sucrose Czapek's agar. The common species on the two types of media used were Penicillium chrysogenum, Aspergillus niger and cladosporium cladosporioides. On 1% glucose Czapek's agar, P. aurantiogriseum, A. fumigatus, A. flavus, Alternaria tenuissima and Pleospora herbarum were isolated with high occurrence while A. versicolor, A. wentii, Eurotium amstelodami and E. chevalieri were common on 40% sucrose Czapek's agar. The different dried fruit samples were assayed for the natural occurrence of aflatoxins B1, B2, G1 & G2, citrinin, ochratoxins, patulin, sterigmatocystin, diacetoxyscirpenol, T-2 toxin and zearalenone by thin layer chromatographic analysis. Ochratoxin A was detected in all samples tested of apricot (50-110 micrograms/kg), fig (60-120 micrograms/kg) and plum (210-280 micrograms/kg). The other mycotoxins under investigation were not detected. All samples examined of raisin proved to be mycotoxin free. PMID- 8229672 TI - Periplasmic aminopeptidases in Acinetobacter calcoaceticus and Pseudomonas aeruginosa. AB - The greater part of the intracellular aminopeptidases in Pseudomonas aeruginosa and Acinetobacter calcoaceticus is soluble. The localization of aminopeptidases in the cells was examined using the osmotic shock method with some modifications. When the cells of A. calcoaceticus and P. aeruginosa of the logarithmic phase were subjected to an osmotic shock, all aminopeptidases investigated were mainly localized in the sucrose supernatants and in the periplasm. Acid phosphatase as marker enzyme for periplasm showed a similar distribution between the fractions as the aminopeptidases. The periplasmic aminopeptidases of both microorganisms were separated by FPLC on Superose 12 and their molecular masses were determined. The results obtained show that at least four different aminopeptidases occur in the periplasm, a leucyl aminopeptidase (LAP, cleaving Leu-NH-NH2, 400 kDa), a glutamyl aminopeptidase (GAP, 200 kDa), an alanyl aminopeptidase (AAP, 80 kDa) and a prolyl aminopeptidase (PAP, 65 kDa). The results are in agreement for both species. Our results show clearly that aminopeptidases of these typical members of Gram-negative bacteria are mainly periplasmic like degrading enzymes (alkaline and acid phosphatases, 5'-nucleotidase, cyclic phosphodiesterase), detoxifying enzymes and binding proteins for amino acids and sugars. PMID- 8229673 TI - Investigations of the oxygen supply in Ca-alginate beads and microcapsules loaded with Penicillium raistrickii using a microelectrode. AB - The oxygen supply of free, Ca-alginate entrapped and microencapsulated mycelia of Penicillium raistrickii i 477 capable of 15 alpha-hydroxylation of 13-ethyl-gon-4 en-3,17-dione was investigated. Using an oxygen microelectrode distinct gradients of oxygen within the Ca-alginate beads as well as the microcapsules were detected. Slope and width of the gradients were investigated in dependence on the kind of immobilization, the culture age and the cell density on or in the carrier as well as the different forms of the oxygen supply in the medium. So it could be shown that large parts of immobilizates, approximately 96% of the diameter of both types, were oxygen-free. In comparison with free mycelia, the lower oxygen supply of the immobilized mycelia led to a metabolic shift to fermentative catabolism. PMID- 8229674 TI - Effect of phenol on the mycelial growth and fructification in some of basidiomycetous fungi. AB - Cometabolic growth studies with phenol were undertaken to screen 32 strains of white and brown rot fungi. All the cultures studied grew well up to 4 mM of phenol on Czapekdox agar except Agaricus bisporus (white button mushroom) and Pleurotus cystidiosus. Most of them could grow even up to 6 mM of phenol. Phenol induced a brown pigmentation of the culture medium. P. flabellatus and P. pulmonarius metabolized 67 and 64 mg/l phenol in 10 days. Studies have indicated that phenol (0.1 to 1.0 mM) incorporated in malt-extract agar has no inhibitory effect on fruitbody formation. Preliminary studies indicate that soaking of wheat straw with phenol solution up to 1600 mg/l give better mycelial growth and fructification of P. cornucopiae, P. ostreatus Z-15 and Calocybe indica than water soaked. Soaking of wheat straw in phenol inhibited the growth of common competitor weed fungi like Stachybotrys sp. and Coprinus sp. PMID- 8229675 TI - Occurrence of antimicrobial activities of bacteria from soybean leaf spots. AB - Bacteria were isolated from leaf spots of field grown soybeans during two growing seasons. The leaf spots yielded up to 4 different species and a total population size of about 10(7)-10(8) bacteria/cm2. The majority of the 192 isolates belonged to the species Pseudomonas syringae pv. glycinea (55%), causing leaf spots of bacterial blight on soybeans, and Erwinia herbicola (22%). The remaining isolates included bacteria from other genera, but occurred occasionally. The determination of biological activity of the isolates demonstrated that a high percentage of strains from the group Erwinia/Enterobacter produced biological active substances against Escherichia coli (69%) and against Chlorella pyrenoidosa (88%). The majority of P. syringae pv. glycinea strains failed to do so. None of the isolates affected the growth of Geotrichum candidum. The E. herbicola strains showed clear antagonistic properties against a wide range of isolated bacteria. Four E. herbicola strains inhibited the growth of nearly all other E. herbicola isolates and 6 other strains were active against most of the P. syringae pv. glycinea isolates. However, antagonistic interactions among strains isolated from a distinct leaf spot were very rarely. PMID- 8229676 TI - Phosphorylated nutrient uptake by Treponema denticola and other potential periodontopathogens. AB - Nutrient uptake may contribute to the survival of certain potential periodontopathogens in the competitive environment of the gingival sulcus. This study was conducted to assess the uptake of commercially available, key radiolabelled phosphorylated compounds by Treponema denticola. There was a linear relationship in the uptake of ATP and G-1-P with cell concentrations of 0.1-2.5 mg of cell dry weight. This uptake reached a steady state within 15 min, and at temperatures of 25-37 degrees C the uptake ranged between 13-21%. At 50 degrees C, or above 50 degrees C the assay cells took up 0-1% of the assay nutrients. The maximum uptake of ATP was 47.5, 38 and 47.6 pmoles for serovars a, b and c. G-1-P yielded maximum uptake values of 306, 304, and 306 pmoles respectively. Significant uptake of ATP, or G-1-P occurred at pH 4.8 for serovars a, or c, while serovar b showed a broad pH range for the uptake of G-1-P. Stannous fluoride at a concentration of 0.17 mM enhanced the uptake of ATP while chlorhexidine digluconate inhibited the uptake of ATP. At concentrations usually employed (0.05-0.5 mM) the metabolic poisons 2,4-dinitrophenol, sodium azide, sodium arsenite and the ATPase inhibitor N,N1-dicyclohexylcarbodiimide did not alter significantly the uptake of ATP. The uptake of ATP, by Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Fusobacterium nucleatum and T. denticola at pH 4.8 was 13.4, 1.5, 6.9, and 18-20.4%, respectively. At pH 6.5 the uptake for the microorganisms indicated above was 22, 1.3, 6.6 and 6.2%, respectively. G-1-P showed the same trend in its uptake by A. actinomycetemcomitans, P. gingivalis, F. nucleatum and T. denticola. This study provides for the first time specific examples of phosphorylated nutrient uptake by potential periodontopathogens, and that the uptake of phosphorylated nutrients can be suppressed by oral hygienic compounds. PMID- 8229677 TI - Stereoselective, competitive, and nonlinear plasma protein binding of ibuprofen enantiomers as determined in vivo in healthy subjects. AB - The plasma protein binding and competitive inhibition parameters of R(-)- and S(+)-ibuprofen were determined in vivo in 12 healthy subjects. Subjects participated in a 4 x 4 Latin square design in which oral solutions of drug were administered as 300 mg R(-)-ibuprofen, 300 mg S(+)-ibuprofen, 300 mg R(-)- + 300 mg S(+)-ibuprofen, and 300 mg R(-)- + 600 mg S(+)-ibuprofen. Unlabeled ibuprofen enantiomers were quantitated using a stereospecific reversed-phase HPLC assay, and plasma protein binding experiments were performed using radiolabeled 14C enantiomers and an ultrafiltration method at 37C. At therapeutic drug concentrations, the protein binding of each enantiomer was greater than 99%. Furthermore, the binding of ibuprofen enantiomers was stereoselective and mutually competitive, as well as nonlinear. The bound-free data were fitted to a model in which the non-linearity of plasma protein binding and competition between enantiomers for binding sites could be accommodated. There were substantial differences in the affinity of ibuprofen enantiomers for protein binding sites (RP2 = 0.358 +/- 0.185 vs. SP2 = 0.979 +/- 0.501 micrograms/ml; mean +/- SD) but no differences in their binding capacity (RP1 = 160 +/- 86 vs. SP1 = 161 +/- 63 micrograms/ml). Although statistically significant, the differences in competitive inhibition parameters were more modest (SKI = 0.661 +/ 0.363 vs. RKI = 0.436 +/- 0.210 micrograms/ml). As a result, the intrinsic binding (i.e., P1/P2) of R(-)-ibuprofen was greater than S(+)-ibuprofen, and the unbound fraction was significantly greater for S-enantiomer vs. R-enantiomer after a given dose of R-ibuprofen or racemate. PMID- 8229678 TI - Nitroglycerin dinitrate metabolites do not affect the pharmacokinetics and pharmacodynamics of nitroglycerin in the dog: a preliminary report. AB - Studies were carried out in conscious dogs to determine the effects of 1,2 glyceryl dinitrate (1,2-GDN) and 1,3-glyceryl dinitrate (1,3-GDN) on nitroglycerin (GTN) pharmacokinetics and pharmacodynamics. In the first set of experiments, steady state plasma levels (Css) of either 1,2-GDN or 1,3-GDN in three dogs were rapidly achieved by giving an iv bolus (77 micrograms/kg), followed immediately by an infusion (50 micrograms/min) of the same GDN. A single iv bolus dose of GTN (0.025 micrograms/kg) was given 50 min after beginning the GDN infusion and compared with plasma concentrations following a similar GTN dose in the absence of dosed GDNs. No significant differences in GTN AUC (p > 0.9) and CL(app) (p > 0.7) were found. In a second set of experiments, an infusion of nitroglycerin was begun in each of 4 dogs and continued for 160 min at an infusion rate of 100 micrograms/min. Steady state concentrations of GTN were achieved within 100 min, at which time the dog received, simultaneously, an iv bolus dose (5.14 mg) of one of the GDNs and an infusion dose (100 micrograms/min) of the same GDN. For both dinitrate metabolites no significant differences (p > 0.5) were found between control and interaction arterial and venous clearances, although venous GTN clearances tended to decrease in the presence of dosed GDNs. Steady state systolic blood pressure during GDN infusions could be further reduced when GTN doses were administered; however, the steady state systolic blood pressure decrease caused by GTN could not be further reduced by the GDN infusions. Results suggest that the GDNs do not inhibit nitroglycerin metabolism or hemodynamics at the dose levels studied here. PMID- 8229679 TI - Effect of phenobarbital and p-hydroxyphenobarbital glucuronide on acetaminophen metabolites in isolated rat hepatocytes: use of a kinetic model to examine the rates of formation and egress. AB - Conventional analysis of initial uptake and egress rates in isolated hepatocytes is limited in the ability to distinguish between rates of metabolite formation and egress, and to separate basolateral and canalicular transport processes. The present study examined the applicability of kinetic modeling in describing acetaminophen glucuronide (AG) and acetaminophen sulfate (AS) formation and egress in hepatocytes after acute exposure to phenobarbital or p hydroxyphenobarbital glucuronide (p-OHPBG) in vitro, or in vivo phenobarbital pretreatment. A significant pretreatment effect on AG and AS disposition was seen based on initial rates of egress. In vivo phenobarbital pretreatment decreased the initial egress rate of AG compared to vehicle pretreatment, and the initial egress rate of AS compared to all other treatments. A pharmacokinetic model incorporating AG and AS formation in hepatocytes as well as egress processes (including diffusional and active transport components) was fit to the data. Parameter estimates derived from model fits to the data showed the expected increase in acetaminophen glucuronidation and decrease in sulfation after phenobarbital pretreatment; in addition, an increase in the AG diffusional rate constant and a decrease in the AS diffusional rate constant was apparent. The excretion Vmax for AG was decreased statistically after acute phenobarbital exposure in vitro, and in vivo phenobarbital pretreatment, with a concomitant statistical increase in the Km for AG excretion. In vitro acute p-OHPBG exposure also decreased significantly the excretion Vmax for AG. These data are consistent with the hypothesis that phenobarbital-impaired biliary excretion of AG is a function of impaired canalicular transport due to the presence of p-OHPBG. They further suggest that the mechanism may not be simple competitive inhibition. This work demonstrates the utility of a kinetic modeling approach to differentiate metabolic and transport processes when analyzing data from isolated hepatocyte studies. Additional information may be gained that would not be apparent by conventional methods of analysis. PMID- 8229680 TI - Application of a simplified method to determine bioavailability of an oral dose of phenytoin. AB - The bioavailability of capsules of phenytoin was determined by two methods: a method involving the numerical integration of the Michaelis-Menten equation and an alternative method involving fitting the time course of plasma concentrations, following the administration of the reference intravenous dosage, to an empirical quadratic function of time. The latter procedure requires much simpler computations. The two methods yielded very similar estimates of the rate and extent of absorption of phenytoin. Total absorption was 0.90 +/- 0.05 and 0.89 +/ 0.05 (mean +/- SE, n = 6) using the methods of numerical integration and quadratic curve fitting, respectively. Both methods indicated that the rate of absorption of phenytoin was inconsistent and slow. Half the total absorption of phenytoin occurred over 2.5 +/- 0.3 hr but the remainder was absorbed very slowly over a period of about 30 hr. Empirical functions may be more generally useful in the determinations of the bioavailability of drugs, particularly if some aspects of the disposition are saturable. PMID- 8229681 TI - Effect of misspecification of the absorption process on subsequent parameter estimation in population analysis. AB - A prospective simulation study has been carried out to evaluate the effect of potential misspecification of the absorption rate constant (ka) in population pharmacokinetic analysis when few to no concentration-time data were available in the absorption phase and estimation of ka was not possible. Data were simulated for 100 subjects using a one-compartment model at steady state with first-order input. Data were generated over a range of ka values: ka was misspecified in the NONMEM analysis by factors of 0.25, 0.5, 1, 2, 3, and 4. In general, clearance (CL) was typically estimated with a small, constant underprediction, regardless of the range of misspecification of ka or whether data were present in the absorption phase. The same was not true for volume of distribution (V), values were biased and sensitive to the degree of misspecification, but only when the data contained even a little information about absorption. If studies are to be designed in which information absorption is either not required or is of no therapeutic use, then blood samples could be concentrated in the postabsorption phase and the absorption input fixed according to the best a priori information available. PMID- 8229682 TI - Targeting the systemic exposure of teniposide in the population and the individual using a stochastic therapeutic objective. AB - A stochastic control approach for dose regimen design is developed and applied to the problem of targeting the systemic exposure, defined as the area under the blood concentration-time curve (AUC), of the anticancer drug teniposide in both the population and individual patients. The control objective involves maximizing the probability that AUC is within a selected target interval given either the population distribution for the kinetic model parameters (a priori control) or the posterior distribution for an individual patient (feedback control). Results of a detailed simulation study are presented, illustrating the feasibility of applying stochastic control principles to the design of dose regimens. The predictive ability of the calculated distributions of AUC for the population and for individuals is evaluated in part by determining the percentage coverage of the computed 95% uncertainty intervals using the simulation results. For the a priori control phase, 94% of the simulated subjects had values of systemic exposure within the computed 95% uncertainty interval, while 93.4% of the simulated subjects had feedback control phase systemic exposure values within their computed 95% uncertainty intervals. Similar evaluation of the uncertainty intervals calculated for plasma concentrations further document the ability of the proposed stochastic control method to predict the uncertainty associated with future therapy. PMID- 8229683 TI - Blood flow measurements in skin and underlying tissues by microsphere method: application to dermal pharmacokinetics of polar nonelectrolytes. AB - Blood flow rates to skin and underlying tissues in rats were determined by the radioactive microsphere technique. The dermal and local tissue clearance of two polar solutes, tritiated water and 14C-labeled sucrose, was assessed in anesthetized and sacrificed rats. The dermal clearance of tritiated water in the presence of viable dermal blood supply (6.17 +/- 1.41 mL/min/100 g) was similar in magnitude but more variable than the blood flow to skin estimated by the microsphere technique (5.18 +/- 0.25 mL/min/100 g). The dermal clearance of sucrose (1.66 +/- 0.32 mL/min/100 g) was approximately one quarter the dermal clearance for water, the difference corresponding to the free diffusion coefficients of the two solutes. The permeability coefficients of tritiated water in various tissues were estimated by both a pseudo-steady-state mass balance approach and numerical integration of differential equations (for dermis only) with a "series compartmental" model, from the tissue concentrations obtained in sacrificed rats. The estimates from numerical integration were comparable to those obtained by mass balance, confirming the general structure of the model. The observed tissue concentrations of tritiated water and sucrose in the anesthetized animal compared well with values predicted from the model. PMID- 8229684 TI - Physiologic models of hepatic drug clearance: influence of altered protein binding on the elimination of diclofenac in the isolated perfused rat liver. AB - The single-pass perfused rat liver preparation was used to assess the influence of binding to human serum albumin on the steady-state hepatic extraction of diclofenac (n = 8). In the absence of binding protein, the extraction ratio of diclofenac approached unity (range, 0.975-0.992), such that its clearance was perfusion-rate limited. As the binding of diclofenac to protein was increased by the addition of human serum albumin to the perfusion medium, its extraction ratio decreased dramatically, and clearance eventually became capacity limited. The relationship between diclofenac availability and fraction unbound was analyzed with various physiologic models of hepatic drug clearance. The dispersion model, which contains a parameter (the dispersion number) that quantifies the axial spreading of a substrate as it passes along the liver length, provided a significantly better description of the data (p < 0.05) than the undistributed parallel-tube model, which assumes that an eliminated substrate travels through the liver as an undispersed plug, and the well stirred (venous equilibrium) model, which assumes that substrate undergoes infinite mixing as soon as it enters the liver. The dispersion number estimated for diclofenac (mean, 3.03; range, 0.89-7.56) was significantly greater than that predicted from considerations of the transverse heterogeneity of blood flow within the hepatic sinusoidal bed, suggesting that additional factors influenced the relationship between availability and fraction unbound for this compound. Such factors may include transverse heterogeneity of the metabolizing enzyme system(s), axial flux of substrate created by diffusion within hepatic tissue, and protein-facilitated transfer of substrate across an unstirred fluid layer adjacent to the hepatocyte surface. PMID- 8229685 TI - Solid-state stereochemistry and activity of 3-methylnefopam diastereomers: manipulation of eight-membered ring conformations in analogues of the non narcotic analgesic drug. AB - The solid-state structures of (+/-)-(1R,3S,5S)/(1S,3R,5R)- and (+)/(-) (1R,3R,5R)/(1S,3S,5S)-3-methylnefopam hydrochloride, epimeric 3-methyl derivatives of the non-narcotic analgesic drug, were determined by single-crystal X-ray diffraction analyses. (+/-)-(1R,3S,5S)/(1S,3R,5S)-3-Methylnefopam hydrochloride gave crystals belonging to the monoclinic space group P2(1)/c, and at ambient temperature, a = 7.993(2), b = 34.376(4), c = 11.785(2) A, beta = 93.06 degrees, V = 3234(2) A3, Z = 8, R(F = 0.070, and Rw(F) = 0.053. (+)/(-) (1R,3R,5R)/(1S,3S,5S)-3-Methylnefopam hydrochloride gave chiral crystals belonging to the orthorhombic space group P2(1)2(1)2(1), and at 92 K, a = 9.261(2), b = 10.280(2), c = 16.668(4) A, V = 1587(1) A3, Z = 4, R(F) = 0.034, and Rw(F) = 0.035. The two molecules in the asymmetric unit of the (1R,3S,5S)/(1S,3R,5R)-racemic modification had twist-chair-(flattened chair) [TCfC] conformational geometries for the eight-membered ring. Both molecules are virtually identical as shown by a root mean squares fit of 0.077 A in the superimposition of all nonhydrogen atoms in both molecules. The (+)/(-) (1R,3R,5R)/(1S,3S,5S)-epimers were found in the same boat-(flattened chair) [BfC] conformation previously noted for crystalline nefopam hydrochloride. The TCfC and BfC eight-membered ring conformations of the two 3-methylnefopam diastereomers differ in the -N+H(CH3)CH2CH-fragment chair or boat arrangement vis-a-vis the adjacent flattened region. In both 3-methyl diastereomers, the C(3)-methyl group was disposed in an equatorial orientation, the phenyl group resided in an exo position, and the -OCH(Ph)-o-C6H4- fragment occupied the flattened region of the eight-membered ring.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229686 TI - Studies on specific inhibition of benzodiazepine receptor binding by some C benzoyl-1,2,3-triazole derivatives. AB - Certain new (1-15) or previously described (16-25) 1,2,3-triazole derivatives, characterized by a C-benzoyl substituent, were synthesized and tested for their ability to displace [3H]flunitrazepam from bovine brain membrane. Compounds 11a and 9a, bearing neutral and lipophilic substituents (phenethyl and cyclohexyl, respectively) showed the higher activity. The 5-benzoyl isomer 11b presented a lower activity, equivalent to that of the triazole acetic derivative 23, which is 4-benzyl substituted. Generally, the carboxymethyl radical in the 1-position of the triazole ring decreased the activity, probably because of intramolecular hydrogen bonding with the carbonyl function of the benzoyl substituent. The N-1 unsubstituted triazole derivatives 24 and 25 were ineffective; this result is in disagreement with our previous observations. Probably these molecules interact with the receptor site by a hydrogen bonding acceptor group and by a bulky and lipophilic portion or a hydrogen bonding donor function that is appropriately arranged. PMID- 8229687 TI - Characterization of the oral absorption of some beta-lactams: effect of the alpha amino side chain group. AB - The intestinal absorption mechanisms of cefixime, 7-aminocephalosporanic acid (7 ACA) and 6-aminopenicillanic acid (6-APA) were determined from the results of single-pass perfusion experiments in rats by modified boundary layer analysis. The estimated absorption parameters (SEM) were as follows: for cefixime, J*max = 0.016 (0.008) mM, Km = 0.031 (0.015) mM, P*m = 0.184 (0.037), P*c = 0.523 (0.051); for 7-ACA, J*max = 6.39 (1.57) mM, Km = 19.33 (5.64) mM, P*c = 0.33 (0.03) mM; and for 6-APA, P*m = 0.41 (0.11), where J*max is the maximal flux of peptide transport system, Km is the intrinsic Michaelis constant, P*m is the dimensionless membrane permeability, and P*c is the dimensionless carrier permeability. Cefixime was absorbed by a carrier-mediated mechanism because its wall permeability (P*w) was concentration dependent and significantly inhibited by cephradine. A concentration-dependent permeability of 7-ACA was observed, but an inhibition study failed to show significant inhibition by cephradine. The absorptions of 6-APA and penicillin V were not inhibited by cephradine or cefixime. The fractions of dose absorbed of several beta-lactam antibiotics correlated well with their absorption numbers obtained from P*w values in rats. These results further demonstrate that an alpha-amino group is not necessary for transport by the intestinal peptide transporter. PMID- 8229688 TI - Hydrogen-bond patterns of dialkylpyridone iron chelators and their 1:1 formic acid solvates: description, prediction, and role in crystal packing. AB - The crystal structures (determined from single-crystal X-ray diffraction studies) and hydrogen-bond patterns of three crystalline 1,2-dialkyl-3-hydroxy-4-pyridones and their 1:1 formic acid solvates are elucidated. The primary hydrogen-bond connectivities observed are explained by a model that predicts that the best donor bonds to the best acceptor. Relative hydrogen-bond donating and accepting abilities of the functional groups observed in these compounds are evaluated by a combination of pKas, energy calculations, resonance arguments, and crystallographic evidence. The primary (O--H ... O) hydrogen-bond patterns are described by graph set notation, and brief explanations of the graph set assignments are also included. A total of 17 secondary (C--H ... O) hydrogen bonds are also observed in these six structures. Correlations are drawn between the observed C--H ... O hydrogen bonds and the molecular packing. The possible role of these secondary hydrogen bonds in influencing molecular packing is discussed. PMID- 8229689 TI - Development of a new drug carrier made from alginate. AB - A new approach for the preparation of nanoparticles is presented. The method is based on control of the gelification phenomenon of alginate by calcium ions, and it leads to small particles of a wide range of very well-defined sizes (250-850 nm) depending on the alignate concentration. The particles are formed in a sodium alginate solution by addition of calcium chloride and then poly-L-lysine. The concentrations of sodium alginate and of calcium chloride were lower than those required for gel formation and corresponded to the formation of a pregel state. The size of the particles formed is greatly dependent on the order of addition of calcium and poly-L-lysine to the sodium alginate solution. This phenomenon can be attributed to the difference in the nature of the interactions between calcium and alginate and between poly-L-lysine and alginate. Furthermore, the data indicate that the formation of the particles probably occurs during the addition of the first component to the sodium alginate solution. Evaluation of the drug loading capacity was done with doxorubicin as a drug model. The results indicate that alginate nanoparticles are interesting carriers because the drug-loading capacity could be > 50 mg of doxorubicin per 100 mg of alginate. PMID- 8229690 TI - Application of neural networks to pharmacodynamics. AB - Neural networks (NN) are computational systems implemented in software or hardware that attempt to simulate the neurological processing abilities of biological systems. A synopsis is presented of the operational characteristics, structures, and applications of NN. The NN technology has primarily been aimed at recognition science (e.g., handwriting, voice, signal, picture, image, pattern, etc.). It is pointed out that NN may also be particularly suitable to deal with pharmacokinetic (PK) and pharmacodynamic (PD) systems, especially in cases such as multivariate PK/PD population kinetics when the systems are so complex that modeling by a conventional structured model building technique is very troublesome. The main practical advantage of NN is the intrinsic ability to closely emulate virtually any multivariate system, including nonlinear systems, independently of structural/physiologic relevance. Thus, NN are most suitable to model the behavior of complex kinetic systems and unsuitable to model the structure. In a practical sense, this structure limitation may be inconsequential because NN in its multivariate formulation may consider any physiologic, clinical, or population variable that may influence the kinetic behavior. The application of NN in PD is demonstrated in terms of the ability of an NN to predict, by extrapolation, the central nervous system (CNS) activity of alfentanil. The drug was infused by a complex computer-controlled infusion scheme over 180 min with simultaneous recording of the CNS effect quantified by a fast Fourier transform power spectrum analysis. The NN was trained to recognize (emulate) the drug input-drug effect behavior of the PD system with the input effect data for the 180 min as a training set.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229691 TI - Reaction of pyrylium salts with nucleophiles. 23: triarylethene derivatives containing an oxyalkyleneamino or oxyalkylene-N-pyridinium side chain. AB - A synthetic design was devised for preparing primary amines related to anticancer drugs clomiphene and tamoxifen on the basis of key intermediates with a phenolic group, to which a side chain (omega-aminoethoxy or omega-aminopropoxy) was attached. These compounds were then reacted with 2,4,6-trimethyl- or 2,4,6 triphenylpyrylium salts. This afforded pyridinium analogues of clomiphene and tamoxifen as potential therapeutic agents for treatment against hormone-dependent tumors. PMID- 8229692 TI - Molecular weight identification of polyethylene glycols in pharmaceutical preparations by gel permeation chromatography. AB - The suitability of different Fractogel media [Fractogel TSK HW40(S), Fractogel TSK HW55(F), and Fractogel TSK HW65(F)] for the molecular weight identification of polyethylene glycols was examined. The results were compared with those obtained on two recently introduced gel filtration media (Superdex 75 prep grade and Superdex 200 prep grade); nonspecific adsorptions were observed for the latter. An aqueous gel filtration procedure was developed for the molecular weight identification of polyethylene glycols in drug formulations as an alternative method to that prescribed by the American Pharmacopeia. This procedure was optimized by a logit-log calibration method. PMID- 8229693 TI - Binding of fluvastatin to blood cells and plasma proteins. AB - The binding of fluvastatin, an inhibitor of hydroxymethylglutaryl coenzyme A reductase, to plasma proteins and red blood cells of rat, dog, and human in vitro was determined by ultrafiltration. Additionally, the stereospecificity of fluvastatin binding to proteins and the potential interaction between fluvastatin and the highly protein bound drugs warfarin, salicylic acid, and glyburide were investigated. Only a small fraction of fluvastatin in blood was taken up by the blood cells, amounting to 19-33% in the rat and < or = 15% in dog and humans. The plasma:blood fluvastatin ratio in these species at 37 degrees C was > or = 1.4. In human blood, this ratio was temperature independent. In the plasma concentration range 25-50,000 ng/mL, fluvastatin was > or = 98% bound to proteins. The binding was concentration dependent in the rat, but not in the dog and human. Both enantiomers of fluvastatin were > 99% bound in normal human plasma, the binding of each being unaffected by the presence of the other. A major fluvastatin-binding protein in human plasma was albumin, whereas binding to alpha 1-acid glycoprotein was relatively weak and concentration dependent. At therapeutic concentrations in normal human plasma, the protein binding of fluvastatin (0.1 microgram/mL) was unaffected by warfarin (1-10 micrograms/mL), salicylic acid (50-150 micrograms/mL), and glyburide (0.1-1 micrograms/mL). Similarly, fluvastatin had no influence on the binding of these compounds. In diluted human albumin solution (29 microM), bound fluvastatin was displaced by all three co-solutes tested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229694 TI - A thermodynamic approach to the binding mechanisms of cefotaxime to serum albumins. AB - The interaction between cefotaxime and the serum albumin of several mammalian species (swine, rabbits, and sheep) at the albumin concentration of 5 x 10(-5) M was studied. Ultrafiltration and spectrophotometric determination of the free antibiotic in the filtrate were used. Binding percentages were determined, and the binding constants and number of sites at various temperatures were calculated. An increase in binding constants, especially in sheep albumin, and an increase in the number of sites from two to three were observed when compared to the results obtained at the physiological albumin concentration. The thermodynamic parameters of interaction varied between the following values: change in Gibbs energy, between -5.7 and -6.4 kcal.mol-1; change in enthalpy, between -6.7 and -9.6 kcal.mol-1; and change in entropy, between -3.1 and -11.2 entropy units. The binding constants increased considerably as the ionic strength of the solution decreased although the number of binding sites remained unchanged. These results indicate the existence of ionic and hydrogen bonds. The possibility of a small contribution from hydrophobic bonds cannot be ruled out. PMID- 8229695 TI - Effect of cimetidine on the pharmacokinetics of fentiazac in rats. AB - The effect of cimetidine (CM) on the pharmacokinetics of fentiazac (FT) was studied in six rats. FT was administered by intravenous (iv) bolus injection at doses of 5, 10, and 20 mg/kg to rats receiving iv infusion of saline (control) or CM. The infusion of CM was conducted to achieve steady-state plasma concentrations (Css) of CM of 30 and 60 micrograms/mL. In the control rats, the plasma disposition of FT and its major metabolite, p-hydroxy FT, followed a classical drug-metabolite profile irrespective of FT dose. The plasma level of p hydroxy FT was comparable with that of FT. The disappearance of FT from the rat plasma was significantly delayed by the CM infusion, and p-hydroxy FT was undetectable in the plasma in CM-treated rats. The total body clearance (CLt) of FT was decreased to 20-50% of the control value in CM-treated rats, implying the FT metabolism to p-hydroxy FT is almost completely impaired by CM. The CLt values in rats with different Css of CM (30 and 60 micrograms/mL) were not significantly different from each other. The distribution volume of FT at steady state (Vdss) was decreased to 60-70% of the control value by the CM infusion at higher doses of FT (10 and 20 mg/kg). The decreases in Vdss and CLt seem not to be related to the binding of FT to plasma protein because the binding of FT was not influenced by CM. In clinical settings, the magnitude of the pharmacokinetic difference may warrant dose adjustment to avoid toxic effects of FT when administered with CM. PMID- 8229696 TI - High-pressure liquid chromatographic assay of theophylline in dog feces following oral administration of sustained-release products. AB - A solid-phase-extraction reversed-phase HPLC assay is described for the determination of theophylline embedded in dog feces as powder, sustained-release tablets, or capsules. The feces is extracted with 5% isopropyl alcohol in chloroform in the presence of beta-hydroxypropyl-theophylline as the internal standard. Separation and quantitation are achieved with a C18 analytical column. UV absorbance is monitored at 280 nm. Recovery of theophylline was > 50%. The assay is linear between 10 and 400 mg amounts of theophylline in 50 g of feces. Inter- and intraday coefficients of variation of the chromatographic assay were < 3%, and the extraction procedure was highly reproducible with coefficients of variation of < 10% at amounts of drug from 10 to 400 mg. By keeping the stool/solvent extraction ratio constant, the method is equally effective in extracting theophylline from different sizes of stool samples (50 versus 200 g of stool). The assay was applied to evaluate the theophylline content in feces following oral administration of the drug to dogs as tablet (Theo-Dur) and capsule (Slo-Bid) dosage forms. The resulting fecal recovery values of each product were inversely related to the corresponding bioavailability values obtained from the literature. PMID- 8229697 TI - Effect of various vehicles on ketoprofen permeation across excised hairless mouse skin. AB - The effects of glycerides, short-chain alcohols, and their binary vehicles as donor components on the permeation of ketoprofen (KP) across the excised hairless mouse skin were evaluated with the modified LOVEDAY-type diffusion cell. Among single vehicles, Panasate 800 as tricaprylin appeared to be the most favorable lipophilic vehicle, with no toxicity and with a short lag time (0.9 h), and methanol (MeOH) or ethanol (EtOH) as hydrophilic single vehicles showed the highest KP permeation flux (244.1 and 134.1 micrograms/cm2/h, respectively). Furthermore, KP permeation was enhanced remarkably by the combination of EtOH and Panasate 800 compared with each single vehicle as reflected in the decreased lag time and increased flux. The greatest enhancement was observed in the EtOH/Panasate 800 (40/60) binary vehicle (permeation ratio at 24 h, 40.0%; steady state flux, 314.0 micrograms/cm2/h; lag time, 3.7 h). Further investigations involving stripping studies and KP accumulation within the skin were performed to explain the mechanism of enhancement caused by the above binary vehicle. It was suggested that the mutual enhancement effect of EtOH/Panasate 800 (40/60) binary vehicle is due to decreasing the barrier ability of the stratum corneum by EtOH and the viable skin by Panasate 800. PMID- 8229698 TI - Constant-rate drug release from novel anionic gel beads with transient composite structure. AB - A new anionic composite bead system with a transient membrane-matrix structure, capable of prolonged constant-rate drug release, has been developed from suspension-polymerized poly(methyl methacrylate-co-methacrylic acid) (PMMA/MAA). These composite beads have a thin PMMA/MAA surface layer and a core consisting of the sodium salt form of the polymer (PMMA/MANa). The high loading (> 20%) of a model drug (oxprenolol HCl) that is achievable in this system from a loading solution concentration as low as 0.5% suggests the formation of a drug-polymer complex in the form of an ionic salt in the core. The release of oxprenolol from such composite beads shows an initial burst effect followed by an extended constant-rate region before leveling off. Apparently, the surface PMMA/MAA layer functions as a transient rate-controlling membrane before it is completely ionized. Because the ionization process is slow, the rate-controlling characteristics of the surface layer and the resulting constant rate of drug release are both sustained for an extended period. The unique feature of the present system is not only its high drug loading capability, but also the transient nature of the rate-controlling surface layer, which is completely ionized towards the latter part of the drug release, thus avoiding prolonged tailing of drug release that is normally associated with permanent membrane matrix systems. PMID- 8229699 TI - Multiple-dose pharmacokinetics of moxisylyte after oral administration to healthy volunteers. AB - The pharmacokinetics of moxisylyte in plasma and urine was investigated after oral administration. Twelve subjects were treated orally, twice daily with 240 mg of the drug for 6 days; on day 7, the subjects received a last dose of 240 mg of moxisylyte. Moxisylyte was assayed in plasma and urine by a specific HPLC method with fluorimetric detection. Moxisylyte was absorbed rapidly and changed to its metabolites immediately after drug administration; unchanged moxisylyte was not found in plasma. Two metabolites were found in plasma and urine: conjugated desacetylmoxisylyte (DAM) and the conjugate of desmethylated DAM (MDAM). The pharmacokinetic parameters determined after the first oral administration were not modified on multiple dosing. The apparent elimination half-lives of conjugated DAM and MDAM were 2.3 and 3.5 h, respectively. Elimination of these two metabolites in urine averaged 50 and 10%, respectively. PMID- 8229700 TI - Determination of total pyridoxal in human plasma following oral administration of vitamin B6 by high-performance liquid chromatography with post-column derivatization. AB - An HPLC method for determining total pyridoxal from plasma was developed for a relative bioavailability comparison of two oral vitamin B6 (pyridoxine HCl) preparations. After cleavage of the pyridoxal-5-phosphate with the acid phosphatase enzyme, the total pyridoxal was determined by HPLC. Pyridoxal was separated on a reversed-phase column, post-column derivatized to pyridoxal semicarbazide, and then detected by fluorescence and quantitated. The limit of detection was 2 ng/mL and interday variation (3 days) over the whole concentration range (13-215 ng/mL spiked) was < 4.1%. In the relative bioavailability study, 16 human subjects were put on a low vitamin B6 diet for a period of 3 days. On the 2nd and 3rd days, 14 blood samples were taken per subject at the same times each day. The drug was administered on the 3rd day. Total endogenous pyridoxal detected on the 2nd day varied in plasma between 13 and 17 ng/mL. Pharmacokinetic parameters corrected for background are reported for two vitamin B6 (40 mg) preparations. Briefly, the pharmacokinetic results for the Ratiopharm preparation compared with the Hoffmann-La Roche preparation are, respectively: AUC0-24, 369.2 and 352.6 ng.h/mL; AUC24-48, 1638.2 and 1662.3 ng.h/mL; net Cmax, 193.0 and 197.1 ng/mL; tmax, 1.25 and 1.44 h; and relative bioavailability, 97.9% (Westlake, 88-112%). PMID- 8229701 TI - Investigation of cell envelope damage to Pseudomonas aeruginosa and Enterobacter cloacae by dibromopropamidine isethionate. AB - Electron micrographs of log-phase Pseudomonas aeruginosa and Enterobacter cloacae cultured for 4 h in the presence of subinhibitory concentrations of dibromopropamidine isethionate indicate that this antibacterial agent can cause marked damage to the cell envelope structures of both species. This result provides an explanation of how dibromopropamidine can enhance the uptake and thus the activity of a second antibacterial agent used in combination with it. PMID- 8229702 TI - Stabilization of aspartame by polyethylene glycol 400. PMID- 8229703 TI - 1993 William J. Stickel Gold Award. Stability of absorbable fixation in basilar first metatarsal osteotomies. AB - The purpose of this study was to compare the structural characteristics of 2.0-mm polyglycolic acid pins and 2.0-mm Steinmann pins in oblique closing base wedge osteotomies of the first metatarsal bone commonly used to correct metatarsus primus varus. Six pairs of fresh frozen cadaveric first metatarsal bones were osteotomized, fixated with either absorbable or stainless steel 2.0-mm pins, cemented in a specimen-holding jig, and tested with the Bionix Material Testing System. There was no significant difference in the mean ultimate force, ultimate displacement, or structural stiffness when comparing 2.0-mm polyglycolic acid pins and Steinmann pins in this model. The structural characteristics of 2.0-mm absorbable pins were the same as stainless steel pins at the time of initial placement in the oblique closing base wedge osteotomy of the first metatarsal. PMID- 8229704 TI - 1993 William J. Stickel Silver Award. Anatomical considerations of the peroneal tubercle. AB - Gross discrepancy and error regarding the identification and location of the peroneal tubercle have been found in the literature. Furthermore, the authors found no evidence of a repeatable measurement technique in applicable descriptions of this osteologic landmark. In accordance with interrater reliability procedures, the authors established repeatability of peroneal tubercle measurements. In conjunction with the instrumentation's accuracy and resolution, the investigators infer reliability for these measurements. To the best of the authors' knowledge, this study is the only one in which interrater reliability was established for the morphometric assessment of the peroneal tubercle and the retrotrochlear eminence. The results also reestablish the correct anatomical presentation of the retrotrochlear eminence and the peroneal tubercle along the lateral surface of the calcaneus. PMID- 8229705 TI - 1993 William J. Stickel Bronze Award. Imaging of the infected foot. Fact or fancy? AB - Advancements in methods of imaging infection have surpassed their worth. Although scientifically substantiated, their clinical pertinence is frequently disappointing. Diabetic infections, postoperative or traumatic sequelae, rheumatologic conditions, and Charcot disease are difficult to differentiate from osseous infection because they all manifest as inflammation. As this country struggles to contain health care costs, a critical investigation regarding the practical benefits of imaging osteomyelitis in the foot is overdue. PMID- 8229706 TI - Aneurysmal bone cyst. AB - Key findings that aid in the diagnosis of an aneurysmal bone cyst are: 1) typically the patients are young individuals less than 20 years old; 2) an expansile bone lesion is bordered by a thin, low signal intensity rim mass; 3) the lesion is inhomogenously increased in signal on T2-weighted images; and 4) multiple fluid-fluid levels are seen with the mass on T2-weighted magnetic resonance imaging scans. PMID- 8229707 TI - Negative bone scan in the diagnosis of calcaneal intraosseous lipoma. PMID- 8229708 TI - Post-iridial implantation revisited. PMID- 8229709 TI - Determination of haptic position of transsclerally fixated posterior chamber intraocular lenses by ultrasound biomicroscopy. AB - Transscleral fixation of posterior chamber intraocular lenses has become an increasingly popular procedure in eyes lacking adequate posterior capsular support. The assumption is generally made that these lenses are fixated in the ciliary sulcus. To test this assumption, 17 cases with transsclerally fixated posterior chamber intraocular lenses were examined with ultrasound biomicroscopy, a new method of producing subsurface images in living eyes at microscopic resolution. All lens haptics were easily visualized with this technique. Of 34 haptics in 17 patients, 13 were adequately located in the sulcus region, eight were located posterior to the ciliary processes, and 13 were located anterior to the sulcus region, accompanied by some degree of angle closure. Haptics with a more posterior scleral exit of the suture tended to be located more posteriorly. The cases in which the haptics were located anteriorly had scleral exit points from 1 mm to 2 mm from the limbus. The surgical placement of transsclerally fixated lenses is a blind procedure in most cases. Our series demonstrates the difficulty in reliably placing the haptics in the ciliary sulcus. PMID- 8229710 TI - Unwanted optical images with narrow profile intraocular lenses compared with conventional intraocular lenses. AB - About 35% of intraocular lenses now implanted in the United States are narrow profile lenses, measuring on the average 5.0 mm by 6.0 mm. This study sought to determine whether these lenses cause unwanted optical images with a decrease in the quality of vision. Seventy-five eyes with a 5.0 mm x 6.0 mm intraocular lens (IOL) were compared with 75 eyes with a 6.5 mm IOL. All patients had capsulorhexis, phacoemulsification, and capsular fixation of the IOL. There was no statistically significant difference in unwanted visual images (16% and 20%, respectively), ability to drive at night (79% and 69%), or patient satisfaction. Careful patient selection is, however, important. PMID- 8229711 TI - Capsule contraction syndrome. AB - Capsule contraction syndrome is an exaggerated reduction in anterior capsulectomy and capsular bag diameter after extracapsular cataract surgery. While rarely seen with can-opener-style capsulectomies with anterior radial capsular tears, it is relatively frequent with capsulorhexis. It is particularly common in patients with pseudoexfoliation and in eyes with a history of moderately severe uveitis. Its effects, which include extreme reduction in the capsulectomy opening, malposition of the opening, reduction in equatorial capsular diameter, and displacement of the IOL, seem more exaggerated in small capsulorhexis openings and in the older patient. Neodymium: YAG laser radial anterior relaxing capsulotomies done within three weeks of cataract surgery reduce the sphincter effect of the contraction and lessen the chronic zonular-traction-related complications of the condition, which may include spontaneous IOL dislocation and retinal detachment. PMID- 8229712 TI - Excimer laser photorefractive keratectomy with tapered transition zone for high myopia. A preliminary report of six cases. AB - To improve the stability of refraction after photorefractive keratectomy (PRK) to correct myopia, we developed a new ablation profile with a 1.5 mm wide tapered transition zone bordering the refractive zone. The treated area's total diameter was 7 mm, the diameter of the refractive zone, 4 mm. The effect of the tapered transition zone on refraction stability was investigated in a 12-month follow-up study of six eyes of six patients. All eyes were partially sighted, with corresponding fundus changes and myopia of between -12.0 diopters (D) and -24.0 D. The therapeutic goal was to reduce myopia by 10.0 D to 13.0 D. All patients were treated unilaterally. The change in refraction in five eyes 12 months after surgery was < or = 1.0 D as compared with the first month postoperatively. In one case the change after 12 months was 2.75 D. There was less regression after PRK with the transition zone than with a conventional ablation profile (i.e., stability was improved). However, epithelial healing took longer. PMID- 8229713 TI - Management of radial keratotomy overcorrections by corneal sutures. AB - Refractive hyperopia is a significant complication after radial keratotomy (RK). In this study, we corrected it by suturing the radial incisions, using purse string sutures, and using a combined technique. Thirteen patients (six original overcorrections, four overcorrections after reoperation, and three progressive hyperopes) were included, demonstrating that the techniques induced a wide range of central corneal steepness and therefore corrected variable amounts of hyperopia. However, only the combination of purse-string suture and radial incision suturing provided a stable result. One patient in the purse-string group and two in the radial group required a third combined surgery to achieve an acceptable result. In the combined group, all cases achieved an improvement in visual acuity. While all cases were within 1 diopter of emmetropia, in seven cases pre-RK visual acuity was equal to the best corrected visual acuity six months after surgery. Preoperative best corrected visual acuity was maintained in all cases. Suture removal at the sixth month postoperatively did not seem to influence the refractive result. Our results suggest that neither purse string nor radial suturing is enough to achieve the girdle effect to the cornea; they must be combined. A significant regression of the effect should be expected following the suture placement, mainly during the early postoperative period, until a stable result is achieved about six months later. A computer-assisted photokeratoscope image was taken preoperatively and postoperatively, showing that some irregular astigmatism should be expected as a result of the procedure. PMID- 8229714 TI - Practice styles and preferences of ASCRS members--1992 survey. American Society of Cataract and Refractive Surgeons. AB - A survey of the practice styles and preferences of the 1992 ASCRS members with a U.S. ZIP code was performed in September 1992. Approximately 38% (1,374) of the 3,582 questionnaires sent out were returned prior to the December cut-off date. Three main profile questions were used to cross-tabulate: the age of the ophthalmologist, the geographic location, and the volume of cataract surgery per month. This report compares the data with previously published surveys of ASCRS members. PMID- 8229715 TI - Fluorophotometry in phakic eyes with anterior chamber intraocular lens implantation to correct myopia. AB - In this longitudinal study, we implanted 16 angle-fixated anterior chamber intra ocular lenses in phakic eyes of 13 patients to correct high myopia. Using fluorophotometry, we discovered a statistically significant decrease (P < .01) in lens transmittance. Blood-aqueous barrier permeability tended to remain increased six months after surgery. PMID- 8229716 TI - Scleral tunnel incision with internal corneal lip in patients with low preoperative corneal endothelial cell counts. AB - We studied 40 eyes from patients with pre-existing corneal endothelial cell dystrophy (endothelial cell counts of less than or equal to 1,200/mm2) to assess whether the scleral tunnel incision with internal corneal lip and phacoemulsification in situ procedure affected the cornea. Identification of endothelial cell dystrophy by measuring preoperative endothelial cell counts was important so minor modifications of decreased phacoemulsification power and frequent viscoelastic endothelial coating could be made to prevent damage to the eyes. The procedures had no effect on corneal thickness or endothelial cell counts. Only about one fourth of the eyes studied (11) had more than 10% endothelial cell loss; none of the 11 eyes showed clinical evidence of corneal decompensation. All patients without other underlying pathologies such as macular degeneration attained postoperative visual acuity of at least 20/40. The scleral tunnel incision with internal corneal lip and phacoemulsification in situ procedure, with minor modifications, is safe in patients with endothelial cell dystrophy of the cornea. PMID- 8229717 TI - Astigmatism after a large scleral pocket incision in extracapsular cataract extraction. AB - In this four-month prospective study, we measured astigmatism in 116 patients who had extracapsular cataract extraction through a 13 mm scleral pocket incision. In one group, the wound was closed with five radial 10-0 polypropylene sutures. Early and late postoperative astigmatism in this group was approximately 2 diopters (D), but a significant shift to against-the-rule astigmatism occurred. In a second group, an additional 10-0 polypropylene horizontal suture was placed anteriorly to the radial suture at the 12 o'clock meridian. This reduced the against-the-rule astigmatic decay. Moreover, this group had less postoperative astigmatism. Mean corneal astigmatism was 2.27 D at one week, 1.76 D at one month, 1.37 D at two months, and 1.70 D at four months. In neither group was cutting the sutures necessary. PMID- 8229718 TI - Astigmatism after phacoemulsification with posterior chamber lens implantation: small incision technique with fibrin adhesive for wound closure. AB - The scleral pocket technique has dramatically changed wound closure after phacoemulsification with implantation of a posterior chamber lens. The use of single-stitch technique and wound closure by fibrin adhesive is now possible. We conducted a comparative study of 385 consecutive patients; 167 received only fibrin glue for wound closure and 218 had the single-stitch procedure. No complications were observed in either group. Surgically induced astigmatism was smaller in the fibrin group (vector analysis: 0.80 diopters [D]) than in the single-stitch group (vector analysis: 0.99 D). Minimal, statistically insignificant different against-the-rule astigmatism developed: single-stitch group: -0.07 D (Cravy), -0.09 D (Naeser); fibrin adhesive group: -0.13 D (Cravy), -0.17 D (Naeser). These results suggest that postoperative against-the-rule astigmatism can be prevented with fibrin glue. PMID- 8229719 TI - Gonioscopy in no-stitch cataract incisions. AB - To evaluate the internal appearance of the no-stitch cataract incision, gonioscopy was performed in 68 of 78 consecutive eyes of cataract surgical patients with no-stitch incisions. All incisions were located anterior to Schwalbe's line, and 32 (42%) had smooth linear internal incisions. In another 32 (42%), small tags or curls were located along the internal incision. Four eyes (5%) had discernible Descemet's membrane detachments visible by gonioscopy but not slitlamp biomicroscopy. The Descemet's detachments were less than 2mm long in three patients and more than 2 mm in one patient. The width of the detachments were 1 mm or less. The detachments did not lead to clinically significant sequela such as filtration blebs or persistent corneal edema. PMID- 8229720 TI - Topical flurbiprofen in extracapsular cataract surgery: effect on pupillary diameter and iris fluorescein leakage. AB - Recent clinical studies indicate that flurbiprofen, a cyclooxygenase inhibitor, prevents miosis and breakdown of the blood-aqueous barrier during cataract surgery. Yet based on clinical and experimental data, some researchers do not agree that flurbiprofen prevents miosis. We conducted a double-blind clinical study of the effects of topical 0.03% flurbiprofen sodium on intraoperative pupillary diameter and iris fluorescein leakage after extracapsular cataract surgery. In the first phase of the study, 120 patients who had extracapsular cataract extraction with posterior chamber intraocular lens implantation were randomly assigned to receive preoperative topical flurbiprofen or a placebo, with or without intraoperative epinephrine, in addition to the standard regimen. In the second phase, 60 of the 120 patients continued the topical flurbiprofen or placebo for one month postoperatively. Iris fluorescein angiography was performed at the end of the first and the fourth weeks. The results indicate that flurbiprofen was significantly more effective (P < .0001) in maintaining mydriasis during surgery than the placebo. This action was enhanced by intraoperative epinephrine. Flurbiprofen also significantly reduced (P < .001) postoperative iris fluorescein leakage. PMID- 8229721 TI - Results of a European multicenter study of the True Vista bifocal intraocular lens. AB - The True Vista intraocular lens (IOL), a three-zone refractive bifocal lens, was implanted in 446 patients in a prospective study. Visual acuity, pupil size, and astigmatism were determined. Contrast acuity was measured in a randomized subgroup of best-case patients (n = 60; Regan charts); subjective assessment of vision was compared in 41 best cases with a monofocal fellow eye. Two hundred twenty-seven best-case patients were available for four to six months follow-up, and 145 for seven to eleven months. Best corrected distance acuity was 20/40 or better in 96% and 98%, respectively. Near acuity was 20/30 or better in 93% and 92% with near add, in 64% and 69% uncorrected, and in 79% and 78% with distance correction, respectively. Distance and near acuity decreased with increasing astigmatism and increasing age. Contrast acuity at distance focus was lower than for monofocal IOLs (11% contrast); values at near focus were considerably lower at all contrast steps tested. Of the patients with a monofocal fellow eye, 96% rated their distance vision with glasses in the monofocal eye as good, compared with 78% in the bifocal eye. The True Vista IOL favors distance vision while near acuity is still sufficient. As high image contrast at distance is usually more important than at near, this might be a good compromise. However, bifocal IOLs should probably not be implanted in patients with a monofocal fellow eye. PMID- 8229722 TI - Comparison of shape recovery ratios of single-piece poly(methyl methacrylate) intraocular lens haptics. AB - We studied shape recovery ratios of several single- and three-piece intraocular lens (IOL) haptics after conducting compression tests for various lengths of time. Results of the comparison of poly(methyl methacrylate) (PMMA) IOL haptics configured of a single piece and of three pieces showed a superior shape recovery ratio for single-piece haptics. Comparisons of the shape recovery ratios among single-piece PMMA haptics and conventional three-piece haptics made of polyvinylidene fluoride (PVDF), PMMA, and polypropylene (PP) revealed that the shape recovery ratio of single-piece PMMA haptics was higher than the ratios of the conventional three-piece PMMA and PP haptics. The ratios of two of three of the single-piece PMMA haptics, however, were lower than the ratios of three-piece PVDF haptics. The form and manufacturing procedures of haptics likely have a significant impact on the shape recovery ratio of haptics, even those made of the same material. Since decentration after IOL surgery could be caused by the condition of the patient's lenticular capsule and Zinn's zonule, it is difficult to conclude that the shape recovery ratio is the only factor in IOL stability. Nevertheless, this study indicates that IOL haptics that cause less decentration and have better mechanical properties should be developed. PMID- 8229723 TI - Decentration associated with asymmetric capsular shrinkage and intraocular lens size. AB - An intraocular lens (IOL) may become displaced even when it has been implanted in the capsular bag. Asymmetric capsular shrinkage is one cause of lens decentration. To determine whether the IOL length influences the degree of decentration, we assessed the decentration of IOLs of different lengths in rabbits. After a D-shaped anterior capsulotomy that covered about half the area of an ordinary circular capsulotomy was performed in 19 eyes of ten white rabbits, a 14.0 mm IOL was fixated in the bag in one eye and a 12.5 mm IOL in the other eye. A circular anterior capsulotomy was performed in 16 eyes of eight controls and IOLs were similarly placed in the bag. Decentration of the 14.0 mm and 12.5 mm IOLs in the eyes with the D-shaped anterior capsulotomy did not differ eight weeks after the procedure. However, marked rotation from the initial position and decentration to the side opposite to that expected were observed in some eyes in which nodular opacification was observed. Both the 14.0 mm and 12.5 mm IOLs were markedly more displaced in eyes with the D-shaped anterior capsulotomy than in eyes with the circular anterior capsulotomy. These results confirm that asymmetric capsular shrinkage, which is affected by the anterior capsulotomy shape, influences post-operative IOL centration. PMID- 8229724 TI - Comparison of efficacy of longest, average, and shortest axial length measurements with a solid-tip ultrasound probe in predicting intraocular lens power. AB - The accuracy of intraocular lens power predictions was calculated for 93 eyes using the shortest, average, and longest of multiple solid-tip probe ultrasound axial length readings for each eye. Averaging the readings reduced the effect of corneal indentation. The single longest and average readings gave comparable results. PMID- 8229725 TI - Efficacy and timed sequence analyses of modified single-injection peribulbar anesthesia. AB - This study evaluated a modified single-injection technique of administering peribulbar anesthesia. All 150 eyes achieved complete lid anesthesia; 49 eyes (32.7%) demonstrating exophthalmos after injection achieved excellent (grade 1) instantaneous ocular akinesia. Sequence of timed events after injection included lid anesthesia (60.2 +/- 15.33 seconds), lid akinesia (75.18 +/- 15.33), lateral rectus akinesia (90.19 +/- 2.13), inferior rectus akinesia (140.44 +/- 17.51), superior rectus akinesia (229.60 +/- 15.23), and medial rectus akinesia (250.42 +/- 18.99). Peribulbar anesthesia, when successful, achieved complete akinesia in fewer than five minutes. In 12 eyes (8%), the peribulbar injection had to be repeated. For routine intraocular surgery, we recommend this efficacious, safe technique without the use of a separate facial block. PMID- 8229726 TI - Ischemic optic neuropathy after intraocular lens implantation to correct high myopia in a phakic patient. AB - A 33-year-old patient had uncomplicated anterior chamber intraocular lens implantation (Worst-Fechner biconcave myopia lens) to correct high myopia. Immediately after surgery, she developed anterior ischemic optic neuropathy likely associated with increased intraocular pressure and systemic hypotension. To our knowledge, this is the first documented case of anterior ischemic optic neuropathy after anterior chamber intraocular lens implantation to correct high myopia in a phakic eye. PMID- 8229727 TI - Spontaneous fistula formation as a complication of relaxing incisions with suture placement for postkeratoplasty high astigmatism. AB - A 39-year-old man had a corneal relaxing incision with corneal compression suture placement to correct more than 10 diopters of corneal astigmatism. A permanent fistula formed from the compression suture, resulting in a localized corneal pocket of aqueous that broke through the surface. The aqueous drained into the tear film, requiring major surgical repair. PMID- 8229728 TI - Consultation section. Degenerative osteoarthritis and systemic hypertension. PMID- 8229729 TI - Hydroexpression of the nucleus. PMID- 8229730 TI - Slow motion phacoemulsification approach. PMID- 8229731 TI - Vector analysis may not apply for astigmatic surgery. PMID- 8229732 TI - Nodal points and capsulotomy size. PMID- 8229733 TI - Inferior congenital iris coloboma and IOL implantation. PMID- 8229734 TI - Infusion misdirection syndrome. PMID- 8229735 TI - The discriminative stimulus effects of cocaine in pigeons. AB - Eight White Carneau pigeons were trained to discriminate 1.0 or 1.7 mg/kg of cocaine from saline. A fixed number of consecutive key peck responses on one key after the administration of cocaine resulted in 4-sec access to mixed grain. The same number of consecutive responses on the other key after saline also produced food. Different doses of cocaine and other drugs were tested to determine their ability to substitute (80% or more responding on the cocaine-appropriate key). The test drugs were selected to determine the selectivity of the cocaine discrimination in pigeons as well the role of different monoamines in mediating this behavioral effect. The drugs included other psychomotor stimulants, antidepressants, clonidine, yohimbine, other dopamine (1-(2-[bis(4-fluoro-phenyl) methoxy]ethyl)4-3-phenylpropyl piperazine, GBR 12909) and serotonin (5-HT, sertraline) reuptake blockers, a D1 (SKF 75670), D2 (quinpirole), and 5-HT1A (8 hydroxy-2-(di-n-propylamino)tetralin, 8-OH-DPAT) agonist as well as the 5-HT3 antagonists, MDL 72222, LY 278584 and ondansetron. In addition, prazosin, an alpha 1 adrenergic antagonist, SCH 23390, a D1 antagonist; raclopride, a D2 antagonist and 1-(2-methoxyphenyl)-4-[4-(2-phthalimmido)butyl]piperazine (NAN 190), a putative 5-HT1A antagonist, were given in combination with cocaine to determine their ability to block the discriminative stimulus (DS) effects of cocaine, i.e., reduce drug-appropriate responding to 20% or less. The psychomotor stimulants, d-amphetamine and d-methamphetamine, completely substituted for cocaine and were similar in potency to each other and cocaine. The antidepressants I-deprenyl, imipramine, tomoxetine and bupropion also occasioned cocaine-appropriate responding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229736 TI - Cardioprotective effects of a novel calcium antagonist related to the structure of cromakalim. AB - A novel pyranoquinoline analog (BMS-188107) of the ATP-sensitive potassium channel (KATP) opener cromakalim was previously shown to be devoid of KATP opening activity in nonischemic myocardium and vascular smooth muscle, but appeared to be a relatively potent calcium antagonist. This clear differentiation between channels within a structural series is a novel finding. With the idea that KATP openers are often more active in ischemic relative to nonischemic myocardium, we determined the cardioprotective effects of this agent in isolated rat hearts and whether these anti-ischemic effects are abolished by KATP blockade. Isolated rat hearts were subjected to 25 min of global ischemia and 30 min of reperfusion and the severity of ischemic/reperfusion injury was determined. BMS-188107 was given before ischemia at 0.5 to 10 microM. Pretreatment (before ischemia) with BMS-188107 caused significant cardiodepressant activity and increased coronary flow only at a concentration of 10 microM, although modest negative inotropic effects were observed at the 0.5 and 1 microM concentrations. Significant improvements in postischemic contractile function and reductions in lactate dehydrogenase release were observed with 1 to 10 microM BMS-188107, indicating significant reductions in ischemic/reperfusion injury. Neither the pre- nor the postischemic effects of 1 to 10 microM BMS 188107 were significantly altered by the KATP blockers sodium 5-hydroxydecanoate (100 microM) or glyburide (1 microM). Previous studies did not determine the effect of BMS-188107 on sodium channels and thus, the effect of this agent on maximum upstroke velocity of the action potential was determined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229737 TI - Combined inhibition of neutral endopeptidase and angiotensin converting enzyme in cardiomyopathic hamsters with compensated heart failure. AB - Inhibition of the metallopeptidase neutral endopeptidase 3.4.24.11 (NEP) protects endogenous natriuretic peptides and potentiates their vasodepressor effects. Inhibition of angiotensin converting enzyme (ACE) attenuates the formation of angiotensin II and enhances the vasodepressor effect of endogenous kinins. A combination of NEP inhibition and ACE inhibition can potentially interact to shift the balance of vasoactive peptides toward vasodilation. This potential interaction was examined in conscious cardiomyopathic hamsters with low cardiac output and compensated heart failure. Neither the selective NEP inhibitor SQ 28,603 nor the selective ACE inhibitor enalaprilat (each at 30 mumol/kg, i.v.) caused significant changes in left ventricular end diastolic pressure or left ventricular systolic pressure when administered separately. However, the combination of these inhibitors, each at that dose, caused significant peak decreases in left ventricular end diastolic pressure and left ventricular systolic pressure of -12 +/- 1 and -18 +/- 4 mm Hg, respectively. In separate cardiomyopathic hamsters, this same combination of treatments resulted in significant decreases in mean arterial pressure (-13%) and total peripheral resistance (-37%) and an increase in cardiac output (36%) as compared with vehicle effects (P < .05). At 90 min after administration of SQ 28,603 alone, plasma atrial natriuretic peptide concentration was double that in the vehicle group. In the group receiving the combination of inhibitors, plasma atrial natriuretic peptide at 90 min was maintained at the high basal levels associated with this model despite the decrease in cardiac filling pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229738 TI - Dose-dependent antagonism and potentiation of nitrous oxide antinociception by naloxone in mice. AB - Administration of the anesthetic gas nitrous oxide (N2O) evoked a concentration dependent antinociceptive effect in mice as assessed by the abdominal constriction test. Depending on the dose and route of pretreatment with the opioid receptor blocker naloxone, the N2O drug effect was either antagonized or potentiated. After s.c. pretreatment with milligram per kilogram doses of naloxone, dose-related antagonism occurred; picogram per kilogram doses potentiated N2O-induced antinociception. The i.c.v. pretreatment with microgram quantities of naloxone also antagonized N2O antinociception in a dose-related fashion; i.c.v. pretreatment with femtogram doses was without effect. On the other hand, intrathecal (i.t.) pretreatment with femtogram quantities of naloxone potentiated N2O antinociception; i.t. pretreatment with microgram quantities continued to antagonize the antinociceptive effect. The same pattern of interaction was observed in mice challenged with the kappa opioid analgesic drug trans (+- 3,4-dichlow-N-methyl-N-[2-(1-pyrrolidinyl)cyclohexyl] benzeneacetamide methane sulfonate (U-50, 488H) after s.c., i.c.v. or i.t. pretreatments with high and low doses of naloxone. These results 1) demonstrate further similarities in the opioid receptor mediation of N2O and U-50, 488H antinociceptive effects and also 2) support the concept of high-affinity spinal opioid receptors, whose blockade by s.c.- or i.t.- but not i.c.v.-administered low-dose naloxone can potentiate the antinociceptive effects of both N2O and U-50,488H. These findings suggest that the antinociceptive effect of N2O might be modulated by a descending opioid system that inhibits analgesia. PMID- 8229739 TI - Effects of antimycin A on the binding properties of beta and alpha-1 adrenergic receptors measured on intact cells. AB - Depletion of intracellular ATP with antimycin A has been shown to inhibit the internalization of several types of receptors, including beta adrenergic and muscarinic acetylcholine receptors. The effects of antimycin A treatment on conversion of beta and alpha-1 adrenergic receptors to a form exhibiting low apparent affinity for agonists in assays with intact cells were investigated, because conversion to the low-affinity form has been postulated to result from receptor internalization. Treatment of DDT1 MF-2 hamster smooth muscle cells with antimycin A greatly decreased conversion of beta adrenergic receptors to the form exhibiting low affinity for agonists. Binding of antagonists to intact cell beta adrenergic receptors was not altered by antimycin A. In contrast to the results with beta adrenergic receptors, conversion of alpha-1 adrenergic receptors to the low-affinity form was not inhibited by antimycin A. These results are consistent with the possible involvement of receptor internalization in conversion to the low affinity form for beta adrenergic receptors. They also provide further evidence that different mechanisms may be involved in conversion to the low affinity form or in the internalization pathway in the case of alpha-1 adrenergic receptors. PMID- 8229740 TI - Role of signal transduction systems in neurotensin receptor down-regulation induced by agonist in murine neuroblastoma clone N1E-115 cells. AB - Murine neuroblastoma clone N1E-115 cells possess neurotensin (NT) receptors, which are coupled to signal transduction systems resulting in polyphosphoinositide (Pl) hydrolysis and cyclic GMP synthesis. Previously, we have demonstrated that the process of down-regulation of NT receptors in N1E-115 cells involves intracellular sequestration of recyclable receptors followed by receptor degradation, causing true down-regulation. In this study, agonist induced sequestration of NT receptors in N1E-115 cells was inhibited by an aminosteroid, 1-(6-([17 beta-3-methoxyestra-1,3,5(10)-trien-17-yl]amino)hexyl)-1H pyrrole- 2,5-diane (U-73122). Pl hydrolysis elicited by NT or sodium fluoride, which stimulates GTP binding proteins, was also inhibited by U-73122, whereas Pl hydrolysis elicited by calcium ionophores, ionomycin or A23187, was not apparently affected. These data suggest that U-73122 affects a process that is distal to the cell surface receptor but not involving the sites just proximal to Pl hydrolysis or cyclic GMP synthesis. It is suggested that U-73122 may affect the coupling of GTP binding proteins and the NT receptor. We conclude that GTP binding proteins play an important role in the mechanism of agonist-induced down regulation of NT receptors in N1E-115 cells. These results may indicate that GTP binding proteins also play a role in the mechanism of internalization of this receptor in the central nervous system in vivo. PMID- 8229741 TI - Systemic excitatory amino acid receptor antagonists of the alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionic acid (AMPA) receptor and of the N-methyl-D aspartate (NMDA) receptor relieve mechanical hypersensitivity after transient spinal cord ischemia in rats. AB - We have previously reported that transient spinal cord ischemia induced a behavioral hypersensitivity (allodynia) to innocuous cutaneous mechanical stimulation in rats. The spinal ischemia-induced allodynia was not relieved by morphine, but it was relieved by the gamma-aminobutyric acid (GABA)-B receptor agonist baclofen, indicating that the allodynia may be related to dysfunction of the spinal GABA-ergic inhibitory system. In the present study we report that systemic application of 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(f)quinoxaline (NBQX), an antagonist of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor for excitatory amino acids, dose-dependently relieved allodynia after spinal cord ischemia. The analgesic effect of NBQX at a low dose (7.5 mg/kg) was not accompanied by motor deficits or sedation. On the other hand, the N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine (MK-801) only partially alleviated allodynia, even at doses that produced severe motor deficits. It is suggested that the abnormal, possibly painful, sensations elicited by innocuous mechanical stimulation observed after spinal cord ischemia may be mediated by excitatory amino acids, acting mainly on the AMPA receptor. Antagonists of excitatory amino acid receptors, especially at the AMPA site, may be effective in treating pain conditions where input from low threshold afferents triggers painful sensations. PMID- 8229742 TI - Cocaine: cardiovascular effects in relation to inhibition of peripheral neuronal monoamine uptake and central stimulation of the sympathoadrenal system. AB - Cocaine (0.03-5.6 mg/kg i.v.) produced a dose-dependent and prolonged increase in mean arterial blood pressure and heart rate in conscious rats. The 0.3 and 1 mg/kg doses of cocaine potentiated the pressor response to exogenous norepinephrine (0.2 microgram/kg), whereas lower (0.03 and 0.1 mg/kg) and higher (3 and 5.6 mg/kg) doses were ineffective. Desipramine (0.03-1 mg/kg), a prototype norepinephrine uptake blocker, did not alter blood pressure or heart rate. Nisoxetine (0.01-1 mg/kg), another norepinephrine selective uptake blocker, produced a small and brief (< 5 min) pressor response, but not tachycardiac response. Unlike cocaine, both desipramine and nisoxetine produced a dose dependent potentiation of the pressor response to norepinephrine with the maximal potentiation exceeding that of cocaine. Plasma norepinephrine and epinephrine levels were increased by cocaine (3 mg/kg), but not by nisoxetine (1 mg/kg). Chemical sympathectomy by 6-hydroxydopamine selectively antagonized cocaine induced increases in both blood pressure and plasma norepinephrine levels, but did not alter cocaine-induced increases in heart rate or plasma epinephrine levels; the converse was the case with adrenal demedullation. Both the combination of chemical sympathectomy and adrenal demedullation and pretreatment with chlorisondamine (10 mg/kg) antagonized cocaine-induced pressor and tachycardiac effects and cocaine-induced increases in plasma epinephrine and norepinephrine levels. In the control group, cocaine (3 mg/kg) produced a biphasic increase in blood pressure consisting of an early peak increase of 52 +/ 2.5 mm Hg 15 sec after its injection followed by a quick and partial recovery to an increase of 20.5 +/- 3.3 mm Hg at 1 min which gradually declined to base-line values in about 30 min. Prazosin (1 mg/kg) pretreatment decreased the magnitude of the initial peak pressor response produced by cocaine and reversed the sustained pressor response to cocaine to a depressor response; the reversal of the pressor response to cocaine to a depressor response by prazosin was not seen after erythro-dl-1-(7-methylindan-4-yloxy)-3-isopropylaminobuta n-2-ol (ICI 118,551) (3 mg/kg) treatment or adrenal demedullation. Treatment with ICI 118,551 alone enhanced the magnitude of the sustained phase of the pressor response to cocaine. These results indicate that inhibition of peripheral sympathetic neuronal amine uptake mechanism by cocaine is not critical for initiating its pressor, tachycardiac and plasma catecholamine increasing effects in conscious rats and that central stimulation of sympathoadrenal neural axis activity plays an important role in these effects.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229743 TI - Anticonvulsant and related effects of U-54494A in various seizure tests. AB - The anticonvulsant activity of (+-)-cis-3,4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl)-cyclohexyl]-benz am ide monohydrochloride (U-54494A), a benzamide derivative chemically related to kappa opioid receptor agonists, was investigated in three selected seizure models of experimental epilepsy. In the maximal electroshock seizure test in mice, U-54494A (ED50 28 mg/kg i.p.) was effective, with a potency somewhat less than phenobarbital. In combination with clinically used antiepileptics, especially phenobarbital and carbamazepine, the anticonvulsant activity of the latter was significantly increased. More detailed studies with phenobarbital showed additive anticonvulsant effects. The anticonvulsant activity of U-54494A was partially antagonized by naloxone. On the other hand, this compound did not elevate the pentylenetetrazol seizure threshold (at high doses a tendency of proconvulsant action was seen). Furthermore, in unrestrained rats with chronically implanted electrodes, U-54494A (> or = 10 mg/kg) significantly reduced the duration of electrically evoked hippocampal afterdischarges. However, the focal stimulation threshold was not markedly increased. With respect to the possible mode of action, whole-cell voltage-clamp experiments on cultured neonatal rat cardiomyocytes showed that U-54494A depressed the fast sodium inward current in a concentration- and frequency dependent manner. In summary, our results agree with earlier reports that demonstrated marked anticonvulsant effects of U-54494A in grand mal-analogous seizure tests. Moreover, in combination with some standard antiepileptics, additive effects can be found. It is suggested that, in addition to kappa opioid and excitatory amino acid receptor related effects, modulations of Na+ membrane currents may contribute to the mechanisms of action. PMID- 8229744 TI - Calcium modulation and cell injury in isolated rat proximal tubules. AB - The possible role of an increased calcium influx in mediating hypoxia-induced injury in isolated rat proximal tubules was examined. Reduction of extracellular calcium delayed the development of proximal tubule cell injury as reflected by the release of lactate dehydrogenase (LDH). This protection, as assessed by LDH release, was not associated with improvements of cell ATP or potassium concentrations. The calcium channel blocker verapamil also delayed the development of cell injury due to hypoxia. This protective effect was evident in the presence of normal or low extracellular calcium. Specifically, hypoxic tubules incubated in low calcium medium were further protected by the addition of verapamil. The protection with verapamil was evident not only by diminished LDH release, but also by improvements in cell ATP and potassium concentration. Further experiments demonstrated that verapamil also increased cell potassium levels in control oxygenated tubules incubated in normal calcium medium. Thus, the results demonstrate that low calcium medium delays the onset of cell membrane injury during hypoxia as assessed by LDH release. The protective effects of verapamil can be demonstrated in the presence of normal or low calcium media, and involve not only diminished LDH release but also increased cell ATP and potassium concentrations. Therefore, in addition to any effect on calcium influx, the effect of verapamil appears to involve a protective effect at cell membrane or mitochondria sites. PMID- 8229745 TI - (+-)-Methyl-2,3,3a,4-tetrahydro-1H-indolo [3,2,1-de] [1,5] naphthyridine-6 carboxylate monohydrochloride facilitates phosphatidylinositol hydrolysis: possible involvement of muscarinic and nonmuscarinic mechanisms. AB - The stimulation of the formation of inositol phosphate (IP) by (+-)-methyl 2,3,3a,4-tetrahydro-1H-indolo [3,2,1-de] [1,5] naphthyridine-6-carboxylate monohydrochloride (vinconate), a novel indolonaphthyridine derivative, was studied using both cerebral cortical slices and crude synaptic membranes prepared from the rat brain. Vinconate (10 mM-1 mM) inhibited the binding of [3H]quinuclidinyl benzilate to the muscarinic receptor in a dose-dependent manner and the IC50 value for [3H]quinuclidinyl benzilate binding was found to be 17 microM. The rightward shift of the inhibition curve of [3H]quinuclidinyl benzilate binding by carbachol in the presence of GTP (100 microM) was abolished by vinconate (100 microM). Carbachol (10 nM-10 mM) significantly increased [3H]IP formation in a dose-dependent manner and the rate of [3H]IP formation mediated by carbachol stimulation was significantly accentuated in the presence of 10 microM vinconate. The enhancement of [3H]IP accumulation by vinconate was inhibited by approximately 50% in the presence of atropine (1-1000 microM), although up to 1 mM of phentolamine and ketanserin had no effect on the vinconate-induced increase of phosphatidylinositol turnover. Moreover, vinconate significantly accentuated 20 mM KCl-evoked stimulation of [3H]IP formation. Vinconate had no differential effect on the ratio of IP or inositol 1,4-biphosphate and inositol 1,4,5 triphosphate formations. These results suggest that vinconate may induce a facilitation of phosphatidylinositol turnover via the stimulation of muscarinic receptors and a facilitation of coupling between muscarinic receptors and GTP binding protein. The presence of a direct stimulatory effect of vinconate on phosphatidylinositol turnover has also been suggested. PMID- 8229746 TI - Central kappa opioid receptor-evoked changes in renal function in conscious rats: participation of renal nerves. AB - The present investigations examined the cardiovascular and renal responses produced by central nervous system stimulation of kappa opioid receptors by the selective kappa opioid receptor agonist, U-50488H, in conscious Sprague-Dawley rats. Administration of U-50488H (1 microgram total) into the lateral cerebroventricle produced a profound diuretic and antinatriuretic response. In addition, concurrent with the decrease in urinary sodium excretion, i.c.v. U 50488H elicited an increase in renal sympathetic nerve activity. The increases in urine flow rate and renal sympathetic nerve activity and the decrease in urinary sodium excretion produced by U-50488H were completely prevented in rats that had undergone pretreatment with the selective kappa opioid receptor antagonist, nor binaltorphimine. In contrast, in animals that had undergone irreversible mu opioid receptor blockade with the selective mu opioid receptor antagonist, beta funaltrexamine, central U-50488H administration elicited similar diuretic and antinatriuretic responses as observed in intact naive animals. In further studies, the antinatriuretic response produced by i.c.v. U-50488H was completely abolished in rats that had undergone chronic bilateral renal denervation, a technique used to remove the influence of the renal sympathetic nerves. Glomerular filtration rates and effective renal plasma flows were not altered by i.c.v. administration of U-50488H in intact or renal denervated animals. Together, these studies provide evidence for the role of central kappa opioid receptor mechanisms in the regulation of urinary sodium and water excretion. Moreover, these studies indicate that the changes in renal sodium handling produced by central kappa opioid agonists result from an action of these compounds to modulate sympathetic neural outflow to the kidneys. PMID- 8229747 TI - Mu- and delta-opioid receptors inhibitorily linked to dopamine-sensitive adenylate cyclase in rat striatum display a selectivity profile toward endogenous opioid peptides different from that of presynaptic mu, delta and kappa receptors. AB - The apparent affinities of endogenous opioid peptides for noncompetitively interacting mu and delta receptors, inhibitorily linked to dopamine (DA) D-1 receptor-stimulated adenylate cyclase, were investigated in superfused rat striatal slices exposed to 40 microM DA in the presence of 10 microM of the selective D-2 receptor antagonist (-)sulpiride. In the presence of peptidase inhibitors, a comparison was made with the apparent affinities of opioid peptides toward independent presynaptic opioid receptors in brain slices. beta-Endorphin had an about 100-fold higher apparent affinity (EC50: 1 nM) toward presynaptic mu opioid receptors, mediating inhibition of the electrically evoked neocortical [3H]norepinephrine release, than for the striatal adenylate cyclase-coupled mu receptors. In contrast, the kappa-opioid receptor agonist dynorphin A1-13 displayed a similar apparent affinity (EC50: 0.1 microM) toward these functionally different mu receptors. Both Leu- and Met-enkephalin showed only a 3 fold higher apparent affinity (EC50: 30 nM) for presynaptic delta-opioid receptors, mediating inhibition of striatal [14C]acetylcholine release, than for presynaptic mu receptors. However, whereas Leu-enkephalin had a similar apparent affinity for presynaptic and adenylate cyclase-coupled delta receptors, Met enkephalin displayed a 30-fold selectivity toward the latter receptors. Studying the inhibitory effect of Met-enkephalin on striatal adenylate cyclase stimulated by endogenously released (amphetamine-induced) DA, its very high affinity appeared to be inversely related to the activation of inhibitory DA D-2 receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229748 TI - Phenytoin blocks N-methyl-D-aspartate responses of mouse central neurons. AB - Intracellularly recorded depolarizing responses of mouse spinal cord neurons in cell culture to N-methyl-D-aspartate (NMDA) applied by pressure ejection at 37 degrees C had a reversal potential of about -13 mV. Amplitude increased when [Mg++]o was less than 1.0 mM or glycine was added to the buffer. Desensitization was complete within 30 pressure applications of NMDA (P30) at 2-s inter-response intervals (IRI; timed from return of one response to resting potential until next application) in bicarbonate buffer and was glycine-sensitive. Desensitization was insignificant in phosphate buffer. In both buffers, 8 x 10(-6) M phenytoin (PT) blocked responses reversibly by P10 of 10(-5) M NMDA at 0.2 Hz (overlapping responses) and at short 2-s IRI (responses not overlapping). At frequencies < or = 0.1 Hz or IRI > or = 5 s, desensitization and block were less prominent or inapparent. Block by PT was observed 1) in single isolated neurons; 2) in 7 mM [Mg++]o-, 150 mM [K+]o-, or tetrodotoxin (TTX)-containing buffer to suppress spontaneous synaptic activity and action potentials and 3) when voltage-dependent Mg++ block was removed by depolarization or in 0.1 mM Mg++, with or without glycine supplementation. The block was not competitive. The PT metabolite, 5-(4 hydroxyphenyl)-5-phenylhydantoin (80 microM), did not block responses to NMDA. Use- and frequency-dependent block of NMDA responses may contribute to clinical effects of PT, e.g., during sustained rapid activity along pathways excited by NMDA-preferring glutamate receptors. PMID- 8229749 TI - Possible novel action of ouabain: allosteric modulation of vascular serotonergic (5-HT2) and angiotensinergic (AT1) receptors. AB - Experiments were carried out to determine the effects of ouabain and dihydroouabain, both at 10 microM, on the vasoconstrictor responses of isolated rabbit blood vessel rings to both serotonin and angiotensin II. Ouabain plus dihydroouabain increased the sensitivity of central ear artery to serotonin and markedly reduced the antagonist potency of 0.1 microM prazosin against serotonin in this blood vessel; dihydroouabain alone had no effect. Ouabain, markedly reversed the reduction of maximal contractile response by both 2-brom-d-lysergic acid diethylamide against serotonin in aorta and 2-n-propyl-4-trifluoromethyl-1 [(2'-1H-tetrazol-5yl)biphenyl -4- yl)methyl]imidazole-5-carboxylic acid (EXP 3892) against angiotensin II in femoral artery; dihydroouabain had no effect. Whereas both 10 microM ouabain and dihydroouabain produced nearly maximal inhibition of Na+K+ATPase, dihydroouabain failed to modulate the vasoconstrictor responses to either serotonin or angiotensin II. Thus, it is concluded that the vasoconstrictor modulatory effects of ouabain are mediated by mechanisms unrelated to Na+K+ATPase inhibition. The present results are consistent with the hypothesis that both 5-hydroxytryptamine2 and angiotensinergic receptors can exist in high and low activity states. It is proposed that 5-hydroxytryptamine2 receptors exist in a low activity state in the ear artery, or are converted to the low activity state by 2-brom-d-lysergic acid diethylamide in the aorta. Hypothetically, 2-n-propyl-4-trifluoromethyl-1- [(2'-(1H-tetrazol-5yl)biphenyl-4 yl)methyl]imidazole-5-carbo xylic acid also converted the angiotensinergic receptor to the low activity state in femoral artery. In all three cases, ouabain enhanced vasoconstriction to serotonin and angiotensin II, respectively, possibly by allosterically modulating the respective receptors in these vessels to their high activity states. PMID- 8229750 TI - Naltrexone-induced upregulation of mu opioid receptors on 7315c cell and brain membranes: enhancement of opioid efficacy in inhibiting adenylyl cyclase. AB - The effect of chronic naltrexone administration on the expression of mu opioid receptors on 7315c tumor cells was examined. Osmotic minipumps containing either saline or naltrexone were subcutaneously implanted into Buffalo rats that had been injected intraperitoneally with 7315c cells. Fourteen days after the pumps were implanted, 7315c tissue and brain tissue were removed and examined for their ability to bind [3H]DAMGO and to respond to morphine (or DAMGO) and guanosine 5' O-(3-thiotriphosphate) in an adenylyl cyclase assay. Naltrexone treatment caused a doubling in the density of [3H]DAMGO binding sites in both whole brain membranes and the 7315c cell membranes. Naltrexone treatment may have slightly diminished the affinity of mu opioid receptors for [3H]DAMGO (by 1.5- to 2-fold), but the precision of the assay was inadequate to determine whether this difference was significant. Naltrexone treatment also had no effect on the potency or efficacy of guanosine 5'-O-(3-thiotriphosphate) in diminishing [3H]DAMGO binding to either whole brain or 7315c cell membranes. The influence of naltrexone treatment on opioid inhibition of adenylyl cyclase activity was also investigated in both tissues. In 7315c membranes, naltrexone treatment caused a 40% increase in the efficacy (maximal effect) of morphine but had no effect on the potency (IC50) of morphine in inhibiting forskolin-stimulated adenylyl cyclase activity. In whole brain membranes from control rats, DAMGO failed to affect significantly forskolin-stimulated adenylyl cyclase. However, in whole brain membranes from naltrexone-treated rats, DAMGO caused a 30% inhibition of forskolin-stimulated adenylyl cyclase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229751 TI - Synergistic effects of glyburide and U-37883A, two structurally different vascular ATP-sensitive potassium channel antagonists. AB - Glyburide, a sulfonylurea, and U-37883A, a guanidine (4-Morpholinecarboximidine-N 1-Adamantyl-N' cyclohexylhydrochloride), have been previously characterized as antagonists of the vascular ATP-sensitive K+ channels (KATP). In this report, the in vitro interaction between these two chemically distinct KATP antagonists was investigated using isolated rabbit mesenteric artery. The KATP antagonism was functionally studied as the inhibition of vasodilation produced by various KATP openers as follows: pinacidil (1 microM), minoxidil sulfate (5 microM), cromakalim (0.5 microM) and RP-49356 (1 microM). Glyburide alone produced inhibition in the concentration range of 50 to 500 nM with the glyburide IC50 ranging from 72 to 148 nM. U-37883A alone produced inhibition in the concentration range of 0.5 to 5 microM, with the U-37883A IC50 ranging from 0.78 to 1.4 microM. In the presence of a threshold U-37883A concentration of 0.5 microM, the glyburide inhibition dose-response curve against pinacidil was significantly shifted to the left and the glyburide IC50 was lowered from 72 to 3.9 nM, representing an 18-fold decrease. Similarly, in the presence of a threshold glyburide concentration of 50 nM, the U-37883A inhibition dose-response curve against pinacidil was significantly shifted to the left and the U-37883A IC50 was lowered from 780 to 96 nM, representing an eightfold decrease. Thus, glyburide and U-37883A potentiated each other's effects as KATP blockers. This synergistic interaction between glyburide and U-37883A was observed independently of the pinacidil, cromakalim or minoxidil sulfate used to produce vasodilation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229752 TI - Oral alcohol self-administration stimulates dopamine release in the rat nucleus accumbens: genetic and motivational determinants. AB - Dopaminergic neurotransmission in the nucleus accumbens may be an important factor in ethanol reinforcement and genetically determined ethanol preference. This hypothesis was tested by measuring dopamine (DA) release by intracranial microdialysis during voluntary oral ethanol self-administration in alcohol preferring (P) and genetically heterogeneous Wistar rats. The animals were trained to respond for ethanol (10% w/v) or water in a free-choice operant task. Extracellular DA levels in the nucleus accumbens were subsequently monitored during 30-min self-administration sessions and a 15-min "waiting period" before session onset. Ethanol self-administration in all animals was followed by a significant, dose-dependent rise in DA release with maximal effects at approximately 15 min after peak intake. Dose-effect functions revealed significantly steeper slopes for the DA-releasing effects of ethanol in P than in genetically heterogeneous Wistar rats. Over an identical range of ethanol doses and blood alcohol levels, increases in DA efflux ranged from 143% to 459% of basal levels in P rats but only from 142% to 212% in Wistar rats. To differentiate the pharmacological effects of ethanol from the effects of operant responding, additional groups of P and Wistar rats were tested during self administration of saccharin (0.05% w/v). By contrast with ethanol, saccharin did not substantially elevate extracellular DA levels. A significant, transient increase in DA efflux was, however, observed in both strains of rats during the presession waiting period in the absence of ethanol or saccharin availability.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229753 TI - Differential vasoconstrictor potencies of free fatty acids in the lung vasculature: 2-versus 3-series prostanoid generation. AB - Pulmonary vasoconstrictor potencies of the 2- and 3-series prostanoid precursors arachidonic acid (AA) and eicosapentaenoic acid (EPA) were compared with each other and three reference fatty acids [palmitic acid (PAL), oleic acid (OA) and eicosatrienoic acid (ETA)]. Dose-effect curves were established from transient pulmonary artery pressor responses (approximately 5-50 mm Hg) evoked by intravascular application of nonesterified fatty acids in buffer-perfused rabbit lungs. Release of di- and trienoic prostanoids into the recirculating perfusate was quantified by a post high-performance liquid chromatography enzyme-linked immunosorbent assay technique. EPA and the three reference fatty acids were used in concentrations up to 10 microM; the rank order of vasoconstrictor potencies was ETA < PAL < OA < EPA. In contrast, AA evoked even larger pressor responses at concentrations two orders of magnitude lower (up to 80 nM). All fatty acids induced both thromboxane A2 and prostaglandin I2 release, ranking with ETA approximately PAL approximately OA < EPA as established for 10 microM concentrations; the dienoic prostanoid release in response to 80 nM AA approximated that elicited by 10 microM EPA. The n-3 fatty acid, however, provoked the liberation of excessive quantities of thromboxane A3 and prostaglandin I3, which surpassed the respective 2-series prostanoids 15- to 20 fold; no 3-series cyclooxygenase products were detected in response to AA, ETA, PAL or OA stimulation. Cyclooxygenase (acetylsalicylic acid) and thromboxane synthetase (OKY 046, Ozagrel, (E)-p-(imidazol-1-ylmethyl)cinnamic acid, C13H12N2O2, MW 228.2) inhibition largely suppressed the EPA-evoked pressor responses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229754 TI - Development of tolerance to the antihypertensive effects of highly selective adenosine A2a agonists upon chronic administration. AB - Three highly A2a-selective adenosine agonists were examined for their effects on blood pressure during chronic administration in conscious spontaneously hypertensive rats. Sodium 4-[2-[[6-amino-9-(N-ethyl-beta-D-ribofuranuronamidosyl) 9H-purin-2 -yl] amino]ethyl]benzenepropionate (CGS 21680C) 2-[(2-cyclohexyl ethyl)amino]adenosine (CGS 22492) and 2-[[2-(1-cyclohexen-1 yl)ethyl]amino]adenosine (CGS 22989) were administered at a rate of 0.25 and 0.5 micrograms/kg/min i.v. for 2 weeks using osmotic minipumps. Significant systolic blood pressure reductions were seen in the A2a agonist-treated groups compared to vehicle-treated (50% dimethyl sulfoxide) animals. Maximum effects occurred on days 1 and 2 in the treated animals. However, the antihypertensive effect diminished with time such that no differences between treatments were seen at 2 weeks. In contrast, a sustained antihypertensive effect was evident with benazeprilat (an angiotensin converting enzyme inhibitor). Tolerance was associated with a decrease in Bmax values (375 +/- 22, 410 +/- 18 and 548 +/- 17 fmol/mg of protein in the CGS 21680C, CGS 22989- and vehicle-treated spontaneously hypertensive rats, respectively) without affecting the Kd value. In addition to a reduction in A2 receptor number, increased heart rates were seen on day 1 and 2 in both the CGS 21680C- and CGS 22989-treated animals and a mild stimulation of the renin angiotensin system occurred with CGS 21680C. In separate acute experiments using identical infusion rates, plasma concentrations of CGS 21680C were 157 +/- 41 ng/ml compared to 30.4 +/- 8.8 ng/ml after chronic administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229755 TI - Effects of cocaine alone and in combination with mazindol in human cocaine abusers. AB - Mazindol is a catecholamine reuptake inhibitor that blocks binding of cocaine at the dopamine reuptake site. This study was conducted to determine whether the acute administration of mazindol modulates the pharmacological effects of intravenous cocaine in humans. In a crossover study, twelve acute drug conditions were tested in randomized order under double-blind, double-dummy conditions in eight cocaine abusers. Cocaine (0, 12.5, 25 and 50 mg, i.v.) was administered in combination with mazindol (0, 1 and 2 mg given orally 2 hr before the cocaine injection). Physiological and subject- and observer-rated responses were measured. Cocaine and mazindol alone both significantly increased heart rate and blood pressure. Cocaine increased ratings on stimulant-like subjective effect measures, including desire for cocaine; mazindol had mild, stimulant-like subjective effects. There were significant interactions between the effects of cocaine and mazindol on heart rate and blood pressure, with combinations producing significantly large and more sustained increases compared with cocaine alone. There was no evidence that mazindol substantially altered the magnitude or profile of the subjective effects of cocaine, including cocaine-induced craving for cocaine. These results do not support the utility of acute administration of mazindol in the treatment of cocaine abusers through a mechanism of modulation of cocaine's subjective effects. Furthermore, mazindol treatment may increase the cardiovascular risks of cocaine. PMID- 8229756 TI - Polychlorinated biphenyl-induced immune suppression: castration, but not adrenalectomy or RU 38486 treatment, partially restores the suppressed cytotoxic T lymphocyte response to alloantigen. AB - The cytotoxic T lymphocyte (CTL) response to allogeneic P815 tumor in C57bl/6 mice is dose-dependently suppressed after treatment with 3,3',4,4',5,5' hexachlorobiphenyl (HxCB). Elevation of plasma corticosterone (CS) is also observed coincident with CTL suppression. Because immune suppression is inducible by glucocorticoid administration, the role of elevated CS was investigated as an indirect mechanism of HxCB-induced immunotoxicity. In multiple experiments, HxCB treatment (10 mg/kg b.w.) consistently reduced CTL activity by 70 to 85% in male mice. Adrenalectomy failed to alter the suppression of CTL activity by HxCB. However, the mortality rate was high (> or = 70%) in these experiments and plasma CS elevation persisted in HxCB-treated adrenalectomy survivors. Therefore, the use of adrenalectomized mice was inadequate to determine whether CS elevation leads to CTL suppression after HxCB treatment. Daily administration of the glucocorticoid receptor antagonist 17-beta-hydroxy-11-beta-(4 dimethylaminophenyl)-17-alpha-(propanyl )-estra- 4,9-dien-3-one (RU 38486) (150 mg/kg b.w., p.o.) also failed to alter the suppression of CTL activity in HxCB treated mice; however, spleen cellularity was significantly increased, suggesting functional GCR antagonism. Male mice were more sensitive to HxCB-induced CTL suppression than female mice, and HxCB-induced plasma CS elevation was greater in male mice. Castration failed to reduce the elevation of plasma CS in HxCB-treated male mice. However, castration partially alleviated CTL suppression in HxCB treated male mice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229757 TI - Platelet-activating factor (PAF) induces a contraction of isolated smooth muscle cells from guinea pig ileum: intracellular pathway involved. AB - This study was designed to evaluate the effect of platelet-activating factor (PAF) on isolated smooth muscle cells from guinea pig ileum circular layer, to characterize the PAF receptors involved in this effect and to determine the intracellular pathways triggered by PAF. Cells dispersed by enzymatic digestion were incubated for 30 sec in the presence of PAF and fixed by glutaraldehyde. When inhibitors or antagonists were tested, cells were preincubated with them for 1 min. Then PAF was added for 30 sec, and the cells were fixed. Contraction was assessed by measuring the length of 50 cells and was expressed as the percentage decrease in cell length from controls. The relaxing effect of inhibitors was expressed as the percentage of the maximal contraction observed in their absence. PAF induced a cell contraction in a concentration-dependent manner. Maximal contraction (24.2 +/- 4.2%) was obtained for a PAF concentration of 10 nM (EC50 = 10 pM). PAF-induced contraction was inhibited by the PAF receptor antagonists BN52021, L659.989 and SR27417. Contraction induced by 10 nM PAF was inhibited when cells were incubated in Ca(++)-free medium with or without 2 mM EGTA or in a 1 mM Ca++ medium to which 100 nM nifedipine was added. When cells were preincubated with concentrations ranging from 0.01 pM to 10 microM of relaxing agents (vasoactive intestinal polypeptide, forskolin, 8 Bromo cAMP) known to increase the intracellular level of cAMP, PAF-induced contraction was inhibited. Moreover, when cells were preincubated with pertussis toxin (200 ng/ml) or cholera toxin (8.4 ng/ml), contraction induced by PAF was also inhibited.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229758 TI - Mechanism of ochratoxin A-induced reduction of glomerular filtration rate in rats. AB - The nephrotoxin ochratoxin A (OTA) causes a reduction of glomerular filtration rate (GFR) and of para-aminohippuric acid (PAH) clearance. We determined renal plasma flow (RPF), filtration fraction (FF) and total renal vascular resistance (TRVR). Using furosemide, we investigated the role of tubuloglomerular feedback (TGF) for GFR reduction. Using enalapril and the angiotensin II antagonist DUP 753 we investigated the role of angiotensin II for GFR reduction. Six days' application of 0.5 mg/kg b.wt. of OTA i.p. to male Wistar rats leads to RPF reduction from 3.98 +/- 0.18 to 1.97 +/- 0.18 ml/(min.g kidney wet weight) and GFR reduction from 1.08 +/- 0.06 to 0.69 +/- 0.04 ml/(min.g kidney wet weight) (n = 6, P < .05 for both). FF increased to 130% of control and TRVR to 202% of control (n = 6, P < .05 for both). The application of furosemide (30 mg/(h.kg b.wt.) after pretreatment with OTA had no effect on the reduction of GFR indicating that TGF is not involved. Pretreatment with OTA and enalapril [0.5 mg/(d.kg) b.wt.] blunted the effect of OTA alone significantly (GFR only dropped to 88% of control). Pretreatment with OTA and DUP 753 [20 mg/(d.kg) b.wt.] blunted the effect of OTA alone significantly: GFR and RPF only dropped to 89 and 91% of control, respectively. FF and TRVR were no longer different from control. Acute application of DUP 753 (2 mg/kg b.wt. i.v.) after pretreatment with OTA blunted the effect of OTA to a lesser extend. Our conclusions are: 1) Reduction of PAH clearance and of GFR is in part due to reduced RPF. 2) The increase in TRVR is at least in part caused by an increase of the efferent resistance. 3) Activation of the TGF is not involved in GFR reduction. 4) The increase in TRVR and the decrease of GFR are mainly mediated by angiotensin II. PMID- 8229759 TI - Discriminative-stimulus effects of the low efficacy mu agonist nalbuphine. AB - The discriminative stimulus effects of nalbuphine were studied in 15 male Sprague Dawley rats trained to discriminate 3.2 mg/kg of nalbuphine from saline under a fixed-ratio 15 schedule of food delivery. Cumulative doses of nalbuphine produced nalbuphine lever responding at doses of 1.0 to 10 mg/kg and rate-suppressing effects at doses of 3.2 to 32 mg/kg. Experiments to evaluate the contribution of opioid receptor activity suggested that the stimulus effects of nalbuphine were mediated through mu systems, inasmuch as mu agonists (etorphine, fentanyl, morphine, buprenorphine, GPA 1657 and nalorphine) produced nalbuphine lever responding, whereas kappa agonists [EKC and U-50,488H (trans-3,4-dichloro-N methyl-N-[2-(1-pyrrolidinyl) cyclohexyl] benzeneacetamide methanesulfonate hydrate)] and nonopioids (d-pentazocine, d-amphetamine and ketamine) produced saline lever responding. dl-Pentazocine produced nalbuphine lever responding in one-half the rats tested. Both high and low efficacy agonists produced nalbuphine lever responding, but the antagonist naltrexone produced predominantly saline lever responding. Increasing the training dose of nalbuphine by a 0.50 log unit failed to alter the potency of nalbuphine or any other compound to produce nalbuphine lever responding, suggesting that these training doses produce a maximum amount of stimulation at the mu receptor. Naltrexone antagonized the discriminative stimulus but not the rate-decreasing effects of nalbuphine, suggesting that only the discriminative stimulus effects of nalbuphine are mediated by a mu opioid mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229760 TI - Complementary and synergistic antinociceptive interaction between the enantiomers of tramadol. AB - The explanation for the co-existence of opioid and nonopioid components of tramadol-induced antinociception appears to be related to the different, but complementary and interactive, pharmacologies of its enantiomers. The (+) enantiomer had Ki values of only 1.33, 62.4 and 54.0 microM at mu, delta and kappa receptors, respectively. The (-) enantiomer had even lower affinity at the mu and delta sites (Ki = 24.8, 213 and 53.5 microM, respectively. The (+) enantiomer was the most potent inhibitor of serotonin uptake (Ki = 0.53 microM) and the (-) enantiomer was the most potent inhibitor of norepinephrine uptake (Ki = 0.43 microM). Basal serotonin release was preferentially enhanced by the (+) enantiomer and stimulation-evoked norepinephrine release was preferentially enhanced by the (-) enantiomer. The (+) and (-) enantiomers each independently produced centrally mediated antinociception in the acetylcholine-induced abdominal constriction test (ED50 = 14.1 and 35.0 micrograms i.t., respectively). Racemic tramadol was significantly more potent (P < .05) than the theoretical additive effect of the enantiomers (antinociceptive synergy). Synergy was also demonstrated (P < .1) in the mouse 55 degrees C hot-plate test (i.p. route) and (P < .05) the rat Randall-Selitto yeast-induced inflammatory nociception model (i.v. and i.p. routes). Critically, the enantiomers interacted less than synergistically in two side-effects of inhibition of colonic propulsive motility and impairment of rotarod performance. The racemate and the (+) enantiomer were active in a chronic (arthritic) inflammatory pain model. Taken together, these findings provide a rational explanation for the coexistence of dual components to tramadol-induced antinociception and might form the basis for understanding its clinical profile. PMID- 8229761 TI - Characterization of [125I][Ser1]histogranin binding sites in rat brain. AB - The binding characteristics of histogranin (HN), an endogenous peptide first recognized for its antagonism of N-methyl-D-aspartate (NMDA) responses, were determined in membrane preparations of rat brain. [125I][Ser1]HN, a stable bioactive analog of HN, bound specifically and reversibly to a homogenous population of high-affinity sites with a Kd of 25 nM and a Bmax of 410 fmol/mg protein. The binding of [125I][Ser1]HN increased linearly with membrane protein concentration and was destroyed upon membrane pretreatment with trypsin. The binding displayed rapid association and dissociation kinetics and was blocked by peptides possessing close homology with HN in the following order: [Ser1]HN-(1 15) > HN > [Ser1]HN-(1-14) > HN-(2-15) > [Ser1]-HN-(1-10) > HN-(6-10). Unrelated peptides such as substance P, beta-endorphin, neuropeptide Y, [Met5]enkephalin, [Leu5]enkephalin, dynorphin A(1-13) and neuromedin C were inactive in competition binding assays against [125I]Ser1]HN. Ligands of the binding domains of the NMDA receptor, such as (+)3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid, (+) 5 methyl-10,11-dihydro 5H-dibenzo[a, d]cyclohepten-5,10-imine hydrogen maleate, 1-N (2-thienyl)cyclohexylpiperidine, glycine and glutamate were also ineffective in competing for [125I][Ser1]HN binding sites. Interestingly, specific ligands for the polyamine site on the NMDA receptor, as well as the cations Mg++ and Zn++ inhibited [125I][Ser1]HN binding. The polyamine antagonist diethylenetriamine produced a noncompetitive inhibition with an IC50 (175 nM) comparable to that of HN (75 nM). The cations Zn++ and Mg++ displaced [125I][Ser1]HN binding with IC50 values of 18 and 240 microM, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229762 TI - Hepatic and splenic phagocytosis in female B6C3F1 mice implanted with morphine sulfate pellets. AB - Morphine sulfate has previously been shown to produce a dose-dependent decrease in hepatic phagocytosis when administered as 8-, 25- and 75-mg pellets implanted subcutaneously. This study was undertaken to determine the time course of suppression of hepatic and splenic phagocytosis after subcutaneous implantation of morphine sulfate pellets. Mice were implanted with either 75 mg of morphine sulfate or placebo pellets. The uptake of chromated sheep red blood cells by the liver and spleen was taken as an index of phagocytosis by resident Kupffer cells or macrophages, respectively. The results indicate that maximum suppression of hepatic phagocytosis by 67% occurred 18 hr after implantation of 75 mg of morphine sulfate. Hepatic phagocytic capacity returned to control levels within 48 hr of implantation. The initial time course of suppression of splenic phagocytosis by 41% was similar to that of the liver (maximum at 12 hr). However, splenic phagocytic capacity returned toward placebo levels over a longer period of time reaching control after 4 days after implantation. The opiate receptor antagonist, naltrexone (30-mg pellet), completely blocked the ability of morphine to suppress either hepatic or splenic phagocytosis. Corticosterone is known to increase in parallel with plasma morphine levels presumably through a hypothalamic-pituitary-adrenal axis. The glucocorticoid receptor antagonist RU 486 was used to block the actions of corticosterone and investigate its possible role in morphine sulfate-induced suppression of phagocytosis. RU 486 (200 mg/kg) completely blocked morphine sulfate's ability to suppress splenic phagocytosis. In contrast, RU 486 only partially blocked morphine-induced suppression of hepatic phagocytosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229763 TI - Urea transport in toad skin (Bufo marinus) AB - Urea is transported from mucosa to serosa across the skin of the stenohaline toad, Bufo marinus, studied under short circuit current (SCC) conditions. Mucosal to serosal transepithelial urea transport (Jm-->s(urea)) was markedly and asymmetrically enhanced in toads adapted to hypertonic (150 mM) NaCl and showed saturation kinetics with an estimated Kd for urea in the bathing solution of approximately 1 mM and a maximal rate of Jm-->s(urea) = 9.4 nmol.cm-2 x hr-1, consistent with a carrier-mediated transport mechanism. Jm-->s(urea) in the skin of 150 mM NaCl-adapted toads was characterized with drugs known to affect transepithelial urea transport (J(urea)) in the urinary bladder of this species. Amiloride (10(-8)-10(-3) M) inhibited Jm-->s(urea) in a dose-dependent fashion, but with a potency only 1/1000th of that for inhibition of SCC in the same skins. Phloretin (< or = 5 x 10(-4) M) had no effect on Jm-->s(urea) or SCC; ouabain (5 x 10(-4) M) and NaCN (10(-3) M) had no effect on Jm-->s(urea) but inhibited SCC (indicating inhibition of active sodium transport) by 70 and 67%, respectively and vasopressin (10(-8) M) had no effect on Jm-->s(urea), but stimulated SCC 179% above base line. The pyrazinoyl amiloride analog, 2-pyrazinoylguanidine (10(-4) M), reported to inhibit urea transport in mammals, also had no effect on Jm- >s(urea), but inhibited SCC approximately 30%. A 1.5 unit pH gradient (m-->s or s ->m) had no effect on Jm-->s(urea).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229764 TI - Inhibition by sulfhydryl compounds of vascular relaxation induced by nitric oxide and endothelium-derived relaxing factor. AB - On rings of rabbit thoracic aorta precontracted with phenylephrine, L-cysteine (Cys) and dithiothreitol (DTT) (1-100 microM), but not glutathione (GSH), produced dose-dependent augmentation of contraction in endothelium-intact, but not in endothelium-denuded rings. The augmentation appeared to be due to inhibition of basally released endothelium-derived relaxing factor (EDRF), and was abolished by pretreatment with superoxide dismutase (SOD, 15 U/ml). At a high dose (1 mM), Cys and GSH produced transient, and DTT produced sustained endothelium-independent relaxation, not influenced by SOD. Cys and DTT (10 microM), but not GSH, produced a small but significant inhibition of acetylcholine-induced endothelium-dependent relaxation, and this inhibition was prevented by SOD. In a perfusion-bioassay system in which EDRF was released by acetylcholine from endothelium of a perfused segment of rabbit aorta, Cys and DTT (20 microM), but not GSH, infused into the perfusate between the segment and an endothelium-denuded bioassay ring, partially inhibited relaxation by the EDRF, but not when SOD was present. In organ chamber experiments, the large transient relaxation of endothelium-denuded rings produced by 75 nM nitric oxide (NO) was partially inhibited in a concentration-dependent manner by Cys, DTT and GSH (0.1 100 microM). Moderate relaxation by 15 nM NO was almost completely inhibited by each compound at 10 microM. The order of potency was Cys > DTT > GSH. Cystine, glutathione disulfide and alanine did not inhibit. Inhibition of NO-induced relaxation was largely attenuated by SOD.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229765 TI - [3H]dopamine and [3H]serotonin release in vitro induced by electrical stimulation in A9 and A10 dopamine regions of rat brain: characterization and responsiveness to cocaine. AB - The present study investigated [3H]dopamine (DA) and [3H]5-hydroxytryptamine (5 HT) release evoked by electrical stimulation in superfused ventral mesencephalon (VM) slices containing A9 and A10 DA neurons. Electrically induced [3H]DA release from VM was, at least, of two origins: one was from DA somatodendrites, regulated by DA autoreceptors, and increased by DA uptake blockers; another was from 5-HT terminals, modulated by 5-HT autoreceptors, and could be minimized by the copresence of fluoxetine during the labeling of the slices. Release of both origins was Ca(++)-dependent and tetrodotoxin-sensitive. Cocaine (10 and 100 microM) modestly increased electrically induced [3H]DA release from DA somatodendrites and concentration-dependently inhibited that from 5-HT terminals. Electrically induced [3H]5-HT release from VM was exclusively from 5-HT terminals, dependent on Ca++, partly blocked by tetrodotoxin, increased by 5-HT uptake blockers and regulated by 5-HT autoreceptors. The autoregulation of [3H]5 HT release partly counteracted the stimulatory effect of cocaine (10 microM) on [3H]5-HT release. Inhibition of 5-HT uptake by cocaine reduced the effectiveness of 5-methoxytryptamine to suppress electrically induced [3H]5-HT release. No evidence was found to support the notion that postsynaptic 5-HT receptors modulate somatodendritic DA release. 5-HT facilitated spontaneous [3H]DA release from VM via a 5-HT/DA transporter-dependent exchange process and inhibited electrically induced [3H]DA release from 5-HT terminals via a 5-HT autoreceptor mediated mechanism. The dual effects of cocaine on electrically induced [3H]DA release from VM may be attributed to the complex 5-HT/DA and 5-HT autoreceptor/cocaine interactions as well as the relative densities of 5-HT/DA transporters in this region, indicating a possible involvement of the 5-HT system in cocaine's behavioral effects. PMID- 8229766 TI - Age-related changes in norepinephrine release and its modulation by presynaptic alpha-2 adrenoceptors in the rat vas deferens. AB - Our objective was to extend the study of age-related changes in the modulation of norepinephrine release by presynaptic alpha-2 adrenoceptors. The experimental model utilized was the epididymal and prostatic portion of the rat vas deferens, because the two portions differ in age-related changes and pattern of sympathetic neurotransmission. The functional influence of presynaptic alpha-2 adrenoceptors was evaluated by quantifying the potentiating effect of yohimbine on the contraction induced by transmural stimulation. Yohimbine-induced potentiation was much more pronounced in the epididymal portion of young rats and was reduced by aging only in this portion. Potassium-induced release of 3H-norepinephrine was significantly higher in the epididymal than in the prostatic portion, and an age related reduction in 3H-norepinephrine release occurred only in the epididymal portion. However, aging did not change the inhibitory effect of clonidine on the contraction induced by transmural stimulation (0.2 Hz or single pulse) in either portion. Thus the age-related decline in alpha-2 feedback regulation is probably not directly related to changes in prejunctional alpha-2 adrenoceptor activity but may be associated with age-related changes in the amount of norepinephrine that stimulates this receptor. However, the lower overflow of neurotransmitter in the epididymal portion did not translate into age-related changes in the contractile responses to transmural stimulation (2-20 Hz). In the prostatic portion, aging increased the functional neurotransmission, partially because of a reduction in neuronal norepinephrine uptake. The results suggest that age-related changes are specific for each portion and may be associated with the pattern of neurotransmission of the tissue. PMID- 8229767 TI - Antinociceptive activity of intrathecally administered potassium channel openers and opioid agonists: a common mechanism of action? AB - The potassium channel openers could share a common mechanism of action with the opiates in the production of antinociception because both classes of drugs have been shown to enhance potassium efflux. The i.t. administration of the potassium channel openers diazoxide, minoxidil, and lemakalim (BRL38227) produced antinociception as measured in the tail-flick test. The ED50's were 122, 184 and 35 micrograms/mouse, respectively. Minoxidil and lemakalim (BRL 38227) were full agonists, whereas diazoxide was a partial agonist. Diazoxide-induced antinociception was blocked by the potassium channel blockers glyburide, apamin and charybdotoxin. Minoxidil- and lemakalim (i.t.)-induced antinociception was blocked totally by only glyburide. The antinociception produced by the potassium channel openers (i.t.) was blocked differentially by opiate antagonists (i.t.). The antinociceptive effects of diazoxide were blocked by nor binaltorphimine, ICI 174,864 and naloxone. Minoxidil- and lemakalim-induced antinociception was blocked by naloxone and ICI 174,864, but not by nor-binaltorphimine. Naloxone (s.c.) shifted the dose-effect curve for minoxidil to the right in a parallel manner. Morphine-induced antinociception was partially blocked by glyburide and apamin, whereas that produced by DPDPE was blocked totally by apamin. U50, 488H induced antinociception was blocked partially by apamin. The potassium channel openers (i.t.) were not cross tolerant to morphine when measured in the tail flick test. Apamin and glyburide precipitated "withdrawal-like" symptoms in morphine-tolerant mice. The interaction of potassium channel openers and opioids probably does not represent a direct interaction of these two classes of drugs at a similar receptor, but rather may occur via an interaction with a common second messenger system such as calcium. PMID- 8229768 TI - Modulation of superoxide anion generation by manoalide, arachidonic acid and staurosporine in liver infiltrated neutrophils in a rat model of endotoxemia. AB - Early accumulation of polymorphonuclear leukocytes (PMN) in the liver after in vivo exposure to Escherichia coli lipopolysaccharide (LPS) and concomitant in vitro phorbol myristate acetate (PMA)-stimulated superoxide anion (O2-) generation has recently been described in a rat model of endotoxemia. The purpose of this investigation was to study the role of phospholipase A2 (PLA2), arachidonic acid (AA), its metabolites, and protein kinase C (PKC) in the mechanism of PMA-stimulated O2- generation of liver infiltrated PMN as compared to circulating blood PMN. Rat PMN were isolated after a 1.5-h infusion of saline or LPS from the blood (SAL-PMN) or the liver (LPS-PMN), respectively. The following results were observed in both SAL-PMN and LPS-PMN: 1) Inhibitors of cyclooxygenase (indomethacin) and 5-lipoxygenase [eicosatetraynoic acid, WY 50,295 tromethamine and VZ 65, 4-(11-hydroxy-1,9-undecadiin)-brenzcatechin] pathways did not inhibit O2- generation; 2) the potent marine PLA2 inhibitor Manoalide inhibited O2- generation in a dose-dependent manner (IC50 = 0.5 microM); 3) exogenously added AA enhanced PMA-stimulated O2- generation in a time and dose-dependent manner and partially reversed the effect of Manoalide in LPS PMN; 4) staurosporine, a putative PKC inhibitor, blocked PMA-stimulated O2- generation completely in the absence of AA and 79% in the presence of AA. It was concluded that LPS-induced liver sequestration of PMN does not alter the role PLA2, AA and PKC play in PMA-stimulated O2- generation. These findings should have implications on the design of novel therapeutic approaches for the modulation of O2- release in the pathogenesis of LPS hepatotoxicity. PMID- 8229769 TI - ATP-sensitive potassium channels and skeletal muscle function in vitro. AB - Compounds that act at ATP-modulated potassium channels (KATP) were tested in an in vitro model of skeletal muscle ischemia. The extensor digitorum longus muscles were removed from anesthetized rats and placed in tissue baths, and contractions were elicited by electrical field stimulation at 0.2 Hz. During normoxia, the force of contraction gradually decayed to about 55% of the peak over 85 min. None of the KATP openers tested, cromakalim (300 microM), P-1075 (10 microM) and pinacidil (100 microM), affected twitch force during normoxia. However, when the muscles were made anoxic, all three compounds greatly accelerated the loss of function in a concentration-related manner. For example, the cromakalim/vehicle ratios of the area under the force-time curve during anoxia were 0.98 +/- 0.03, 0.77* +/- 0.03 and 0.72* +/- 0.04 for cromakalim at 30, 100 and 300 microM, respectively (*P < .05). Upon reoxygenation, muscles treated with the KATP openers recovered twitch force to a greater extent than those treated with vehicle. Glyburide (1 or 10 microM) had no effect on its own, but it was able to prevent fully the effects of KATP openers during both anoxia and reoxygenation, indicating that the effects of the KATP openers were mediated by KATP. These results suggest that KATP openers would not be beneficial in the treatment of skeletal muscle ischemia in vivo but that they may be useful in preserving skeletal muscle function in cases of ischemia followed by reperfusion. PMID- 8229770 TI - Possible involvement of dopamine in the long-term serotonin depletion by p chloroamphetamine and beta,beta-difluoro-p-chloroamphetamine in rats. AB - The role of dopamine in the long-term depletion of serotonin in rat brain by p chloroamphetamine (PCA) and related compounds was investigated by comparing effects of beta,beta-difluoro-p-chloroamphetamine (beta,beta-difluoro-PCA) and 4 methyl-alpha-ethyl-meta-tyramine (H75/12), reported to cause only short-term serotonin depletion, with those of PCA. A single dose of beta,beta-difluoro-PCA had no long-term effects on serotonin in whole rat brain, even after pretreatment with proadifen which decreased the rate at which beta,beta-difluoro-PCA disappeared from brain. The possibility that proadifen might antagonize serotonin depletion was ruled out; proadifen did not prevent long-term serotonin depletion by PCA. Long-term depletion of brain serotonin was found after repeated injections of beta,beta-difluoro-PCA (five injections 4 hr apart) and was prevented by fluoxetine pretreatment. beta,beta-Difluoro-PCA given after the monoamine oxidase inhibitor pargyline or after carbidopa/L-dopa also caused long term serotonin depletion, although H75/12 did not. At early times after single doses producing the same initial depletion of serotonin, PCA caused a large increase in dopamine and a large decrease in the metabolite 3,4 dihydroxyphenylacetic acid in whole brain, thereby increasing the ratio dopamine/3,4-dihydroxyphenylacetic, and the other two drugs caused smaller effects. Extracellular dopamine was increased markedly by PCA, less by beta,beta difluoro-PCA, and not at all by H75/12. These results suggest an association between dopamine release and long-term depletion of serotonin and add to evidence that dopamine release by PCA may be essential to its neurotoxic actions on brain serotonin neurons. PMID- 8229771 TI - Transplacental and nonplacental clearances, metabolism and pharmacodynamics of labetalol in the fetal lamb after direct intravenous administration. AB - Labetalol has been previously shown to cause significant maternal and fetal metabolic effects in pregnant sheep after maternal administration. To investigate these observations further, the present study describes the pharmacokinetics, metabolism and pharmacodynamics of labetalol in the fetal lamb after direct fetal i.v. bolus (4 mg) administration. The fetal total body clearance of labetalol (50.45 +/- 1.37 ml m-1 kg-1), which was significantly higher than that previously determined in the ewe, was composed of transplacental and nonplacental CLs of 23.4 +/- 8.99 ml m-1 kg-1 and 27.05 +/- 10.36 ml m-1 kg-1, respectively. The maternal to fetal plasma labetalol area under the curve ratio was 0.031 +/- 0.002 and the CLmp and CLmn were 7.27 +/- 2.11 ml m-1 kg-1 and 30.5 +/- 5.94 ml m-1 kg 1, respectively. Labetalol concentrations in fetal tracheal fluid were consistently higher than that in fetal plasma. The glucuronide conjugate of labetalol was found in the amniotic fluid at up to 20 times the free drug concentration but the oxidative metabolite, 3-amino-1-phenyl-butane, was not detected in plasma or amniotic fluid samples. The fetal effect of labetalol was characterized by an acute lactic acidosis. The calculated hind limb arteriovenous lactate flux showed a net output of lactic acid equal to 3.85 +/- 2.05 g from the hind limb over 24 h after labetalol administration. Although the fetal exposure to labetalol in this study was roughly 4 times that after a 100-mg maternal bolus administration, the magnitude of fetal lactic acidosis was not significantly different in these studies. The clinical implications of the observations made in this study remain to be investigated. PMID- 8229772 TI - Effects of CGS 15943, a nonxanthine adenosine antagonist, on behavior in the squirrel monkey. AB - The behavioral effects of CGS 15943 (0.1-3.0 mg/kg), a nonxanthine adenosine antagonist lacking phosphodiesterase (PDE) inhibitory effects, and caffeine (1.0 30.0 mg/kg), a xanthine adenosine antagonist with PDE inhibitory effects, were compared in squirrel monkeys trained to lever-press under fixed interval (FI) schedules of food presentation or stimulus termination. Both adenosine antagonists increased FI response rates after i.m. or i.v. administration, with CGS 15943 being more efficacious and approximately 3 to 10 times more potent than caffeine. Moreover, the rate-increasing effects of caffeine were enhanced by CGS 15943 (0.3 and 1.0 mg/kg) pretreatment. In contrast, rolipram (0.01-0.1 mg/kg), a potent PDE inhibitor lacking adenosine-antagonist effects, only decreased response rates. The nonselective adenosine agonist, 5'-N-ethylcarboxamidadenosine (0.003-0.03 mg/kg), the A1-selective adenosine agonists, N6-cyclopentyladenosine (0.1-1.0 mg/kg) and N6-cyclohexyladenosine (0.1-1.0 mg/kg) and the A2-selective adenosine agonist, CGS 21680 (0.03-0.3 mg/kg), produced dose-dependent decreases in response rates that were attenuated by CGS 15943 and caffeine. The potency difference between CGS 15943 and caffeine as antagonists of 5'-N ethylcarboxamidadenosine, N6-cyclopentyladenosine and N6-cyclohexyladenosine corresponded to the potency difference of the two drugs for increasing FI response rates. In contrast, CGS 15943 and caffeine were approximately equipotent as antagonists of CGS 21680. The similarity of the effects of CGS 15943 and caffeine supports and extends previous findings suggesting that the behavioral stimulant effects of caffeine and other xanthines are mediated through adenosine antagonist actions rather than inhibition of PDE activity. PMID- 8229773 TI - Nicotinic agonists modulate basal forebrain control of cortical cerebral blood flow in anesthetized rats. AB - Previous studies have indicated that electrical microstimulation of the cholinergic (basal forebrain, BF) elicits profound increases in cortical cerebral blood flow (CBF) that are selectively attenuated by nicotinic receptor antagonists. This study sought to determine whether nicotinic receptor agonists such as (-)-nicotine, and related agents, can enhance the increases in CBF elicited by electrical stimulation of the BF of urethane-anesthetized rats. The magnitude of cortical CBF responses, measured by laser-Doppler flowmetry, increased progressively with higher frequencies (range = 6.25-50 Hz) to a maximum of 248% of control. (-)-Nicotine and (-)-lobeline each further enhanced the responses to BF stimulation, with (-)-nicotine having the most potent effect (up to 350%). (+)-Nicotine and (-)-cotinine were without effect, suggesting stereoselectivity and that the effects were not mediated by the major metabolite of (-)-nicotine. In contrast, (-)-cystisine, another nicotinic receptor agonist, modestly inhibited the BF-elicited increase in CBF suggesting nicotinic receptor subtype selectivity in mediating the response. Arecoline, a potent muscarinic agonist, was without effect suggesting that muscarinic mechanisms are not involved in the mediation of this response. None of the nicotinic agents had overt effects on heart rate or blood pressure in the dose ranges examined. In experiments targeting the site of action of the nicotinically mediated enhancement, (-)-nicotine microinjections into the BF elicited profound increases in cortical CBF, whereas similar injections into the cerebral cortex were without effect suggesting that nicotine receptors mediating CBF increases are localized to the BF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229774 TI - Modulation of morphine antinociception by swim-stress in the mouse: involvement of supraspinal opioid delta-2 receptors. AB - The present study evaluated the effect of a brief exposure of mice to cold-water swim-stress (CWSS) on the antinociceptive potency of i.c.v. given morphine. No significant antinociceptive response could be demonstrated in the warm-water tail flick test, 10 min after a 30-sec exposure of mice to water at 5 degrees C. However, the i.c.v. morphine dose-response curve in mice exposed to CWSS was displaced significantly to the left when compared to that obtained in control (i.e., non-CWSS-exposed) mice. Although coadministration of the delta antagonist, N,N-diallyl-Tyr-Aib-Aib-Phe-Leu-OH 1 (ICI 174,864), with i.c.v. morphine did not produce antagonism of the antinociceptive action of this mu opiate, the leftward displacement of the i.c.v. morphine dose-response curve seen in CWSS-exposed mice was blocked in ICI 174,864-treated mice suggesting involvement of opioid delta receptors in the modulatory effect. Pretreatment of mice with the delta-1 antagonist, [D-Ala2, Leu5, Cys6] enkephalin, did not antagonize the antinociception of morphine and further did not antagonize the leftward displacement produced by exposure to CWSS. Pretreatment of mice with the delta-2 antagonist, 5'-isothiocyanate, also did not antagonize the antinociceptive effects of morphine but blocked the leftward displacement in the morphine dose response curve associated with CWSS, suggesting involvement of an opioid delta-2 receptor in this effect. Pretreatment of mice with the mu antagonist, beta funaltrexamine, produced a significant antagonism of the morphine antinociceptive effect as seen by a rightward displacement of the morphine dose-effect curve.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229775 TI - Serotonergic modulation of L-glutamic acid-evoked release of endogenous norepinephrine from rat hypothalamus. AB - Hypothalamic slices (400 mu) from male Sprague-Dawley rats were perfused with a Mg+(+)-free medium containing nomifensine (10 microM) and tyrosine (50 microM). Spontaneous release of endogenous norepinephrine (NE), measured by high performance liquid chromatography-electrochemical detection, averaged 102 +/- 13 (N = 76) fmol/mg of protein/3 min. L-Glutamic acid (L-GLU) (1 mM) more than doubled the rate of NE release. Preincubation with serotonin (5-HT) (0.1-10 microM) produced no change in spontaneous NE release but caused a concentration dependent decrease of L-GLU-induced NE release with a maximal reduction of about 60 to 70%. 2-Methylserotonin, a 5-HT3 receptor agonist (0.07-10 microM), mimicked the 5-HT response. A highly selective 5-HT3 receptor antagonist, (3 alpha tropanyl)1H-indole-3-carboxylic acid ester, 1 nM, inhibited the effect of both agonists. Neither ritanserin (1 microM) nor methylsergide (1 microM) modified either spontaneous or 1 mM L-GLU-evoked release of NE. However, if added to the superfusion medium simultaneously with 5-HT, they potentiated significantly the inhibition produced by 5-HT. Alpha-methylserotonin (1 microM) if added alone to the perfusion medium had no effect on 1 mM L-GLU-evoked release of NE but reversed the inhibition induced by 1 microM 2-methylserotonin. These observations provide direct evidence of a dual modulation by 5-HT of L-GLU-evoked release of endogenous NE from slices of rat hypothalamus: An inhibition mediated by 5-HT3 receptors and an opposing action mediated by receptors of the 5-HT1C/2 type. PMID- 8229776 TI - Effects of nifedipine, 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate hydrochloride and atrial natriuretic peptide on endothelin-induced antinatriuresis in dogs. AB - Nifedipine, 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate hydrochloride (TMB 8) or atrial natriuretic peptide (ANP) was infused into the renal artery before and during intrarenal arterial infusion of endothelin-1 (ET) in anesthetized dogs. Before ET infusion, nifedipine (0.1 micrograms kg-1 min-1), TMB-8 (75 micrograms kg-1 min-1) or ANP (10 ng kg-1 min-1) increased the urine flow rate, urinary sodium excretion and fractional sodium excretion with little change in renal blood flow or glomerular filtration rate. ET (2 ng kg-1 min-1) reduced the basal renal blood flow, glomerular filtration rate, urine flow rate, urinary sodium excretion and fractional sodium excretion. Both nifedipine and TMB-8 induced natriuresis during ET infusion; but only TMB-8 completely reversed the ET induced reduction in fractional sodium excretion and partially antagonized the reductions in urine flow rate and urinary sodium excretion. ANP did not induce substantial urinary responses during ET infusion. Neither nifedipine, TMB-8 nor ANP reversed the ET-induced decreases in renal blood flow and glomerular filtration rate. The present study suggests that in the dog kidney 1) the ET induced antinatriuresis is caused in part by enhancement of tubular sodium reabsorption, 2) the tubular action of ET depends on TMB-8-sensitive calcium movements but not calcium influx through dihydropyridine-sensitive channels and 3) ANP cannot counteract the ET-induced antinatriuresis. PMID- 8229777 TI - Stimulation of brain steroidogenesis by 2-aryl-indole-3-acetamide derivatives acting at the mitochondrial diazepam-binding inhibitor receptor complex. AB - The 2-aryl-indole-3-acetamide derivatives, 2-hexyl-indole-3-acetamide (FGIN-1-27) and 2-hexyl-indole-3-acetamide-N-benzene-tricarboxylic acid (FGIN-1-44) displaced [3H]1-(2-chlorophenyl)-N-methyl-N-(1- methylpropyl)-3-isoquinoline carboxamide([3H]PK 11195) and [3H]4-chlorodiazepam ([3H]4'CD) from binding sites located on the rat brain mitochondrial DBI receptor complex (MDRC) with Ki values in the nanomolar range. Both 2-aryl-indole-3-acetamide derivatives acted as agonists at the MDRC and thereby stimulated the rate of pregnenolone synthesis in isolated rat brain mitochondria; this effect was inhibited by PK 11195, an MDRC ligand that does not possess steroidogenic activity. FGIN-1-27 and FGIN-1-44 failed to bind to other transmitter receptors, including gamma-aminobutyric-A receptors. When administered orally to rats, both FGIN-1-27 and FGIN-1-44 reduced fear of novelty in the elevated plus maze test. This action was prevented by PK 11195, but not by flumazenil. FGIN-1-44, which was rapidly converted to FGIN-1-27 in the rat brain, was 3 to 4 times more potent than FGIN-1-27 in reducing fear of novelty because of its greater bioavailability. FGIN-1-27 increased the brain pregnenolone content in adrenalectomized-castrated rats pretreated with trilostane (in order to prevent metabolism of pregnenolone to progesterone). This increase was blocked by pretreatment with PK 11195. Although FGIN-1-27 and FGIN-1 44 increased the corticosterone concentration in adrenal glands and plasma of hypophysectomized rats in a PK 11195-sensitive manner, both drugs failed to increase adrenal steroidogenesis in sham-operated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229778 TI - Use of m1-toxin as a selective antagonist of m1 muscarinic receptors. AB - m1-Toxin is the only ligand which is known to bind specifically to the extracellular face of genetically defined m1 muscarinic receptors; it binds pseudoirreversibly. A variety of studies were performed to evaluate the usefulness of m1-toxin as a selective antagonist of m1 receptors. Exposure of slices of the rat cerebral cortex to m1-toxin in physiological buffer blocked the subsequent binding of 1.0 nM [3H]pirenzepine to m1 receptors in the slices. The toxin also blocked 70% of carbachol-stimulated turnover of radiolabeled inositol phosphates in hippocampal slices. Autoradiographs showed that m1-toxin bound to sections of once-frozen tissue and blocked the binding of [3H]quinuclidinyl benzilate to regions of the rat brain rich in m1 receptors. The toxin blocked the binding of [3H]antagonists to pure m1 receptors on the surface of living Chinese hamster ovary cells, but did not block intracellular receptors. In membrane preparations from the rat cortex and hippocampus the toxin blocked the binding of [3H] antagonists to m1 receptors quantitatively and selectively, but had no effect on binding sites for [3H]nicotine. Subsaturating amounts of the toxin bound to m1 receptors in membranes at 4 degrees C in less than 30 sec. Low concentrations of m1-toxin blocked m1 receptors in solution in digitonin but had no effect on separate preparations of pure m2, m3, m4 or m5 receptors. Thus m1 toxin appears to be a very useful antagonist for m1 receptors in intact tissue, on isolated cells, in membranes and in solution, in a variety of media. PMID- 8229779 TI - Involvement of N-methyl-D-aspartate receptor stimulation in the ventral tegmental area and amygdala in behavioral sensitization to cocaine. AB - Systemic administration of N-methyl-D-aspartate (NMDA) antagonists prevents the development of behavioral sensitization to amphetamine-like psychostimulants. Pretreatment with the noncompetitive NMDA antagonist, MK-801, resulted in a dose dependent blockade of behavioral sensitization to cocaine. However, pretreatment with the highest dose of MK-801 (0.25 mg/kg i.p.) alone inhibited the behavioral response to a subsequent cocaine challenge 24 hr later. The induction of behavioral sensitization is known to result, at least partly, from an action by psychostimulants in the ventral tegmental area (VTA). To determine whether the dose-dependent inhibition of behavioral sensitization to cocaine by NMDA antagonists resulted from receptor blockade in the VTA, rats were pretreated in the VTA with the MK-801 or the competitive NMDA antagonist, 3-(2 carboxypiperazine-4-yl) propyl-1-phosphonic acid, before systemically administered cocaine (30 mg/kg i.p.). Two to 3 days later rats were challenged with cocaine alone (15 mg/kg i.p.). Pretreatment with either NMDA antagonist into the VTA prevented the manifestation of behavioral sensitization. Intracranial pretreatment with MK-801 was also made into the nucleus accumbens and amygdala which have been implicated in psychostimulant-induced sensitization. Whereas MK 801 was without effect in the nucleus accumbens, when microinjected into the ventral amygdala it prevented the manifestation of behavioral sensitization to a cocaine challenge. The blockade of sensitization by MK-801 in the VTA was produced with a minimum effective dose of 0.01 nmol, whereas the minimum effective dose in the amygdala was 1.0 nmol. These data demonstrate that stimulation of NMDA receptors in the VTA and amygdala is necessary in the development of behavioral sensitization to cocaine. PMID- 8229780 TI - Activation of transcription factor genes in striatum by cocaine: role of both serotonin and dopamine systems. AB - Acute administration of cocaine increases expression of the transcription factor genes c-fos and zif268 in the striatum. This response is thought to be mediated via D1 dopamine (DA) receptors, as it is blocked by the selective D1 receptor antagonist SCH 23390. However, the directly acting D1 receptor agonists, apomorphine and SKF 38393, do not mimic cocaine's activation of these genes raising the possibility that D1 receptor activation is necessary, but not sufficient, to trigger transcription factor expression. Because cocaine blocks uptake of norepinephrine (NE) and serotonin (5-HT), as well as DA, we examined whether cocaine's ability to inhibit NE and 5-HT uptake may contribute to its induction of c-fos and zif268 expression in striatum. In examining the effects of selective monoamine uptake inhibitors, we observed that fluoxetine or citalopram, selective inhibitors of 5-HT uptake, potentiated the ability of mazindol, a DA and NE uptake inhibitor, to induce zif268 and c-fos expression, even though these 5-HT uptake inhibitors had no effect when administered alone. In contrast, the selective NE uptake inhibitor, desipramine, administered alone, or in combination with fluoxetine, did not increase expression of zif268 or c-fos. Furthermore, selective denervation of 5-HT projections by p-chloroamphetamine treatment attenuated the increase in zif268 and c-fos expression induced by cocaine in the striatum. In contrast, selective lesions of NE projections with N-(2-chloroethyl) N-ethyl-2-bromobenzylamine hydrochloride failed to block cocaine's activation of these genes in the striatum. Taken together, these findings indicate that cocaine's ability to induce striatal expression of c-fos and zif268 is mediated by its effects on both the 5-HT and DA systems. PMID- 8229781 TI - Effects of intracerebroventricular administration of beta-funaltrexamine on [3H]DAMGO binding to rat brain sections. AB - In spite of an extensive body of knowledge regarding the pharmacological effects of centrally administered beta-funaltrexamine (beta-FNA), little is known about the distribution of mu opiate receptor alkylation produced by i.c.v. administration. This study examines the dose relationship between i.c.v. beta-FNA pretreatment and the affinity and density of mu opioid binding sites in discrete brain regions using in situ binding and quantitative receptor autoradiography. [3H]DAMGO binding was determined in coronal sections obtained from the frontal portion of the brain from animals 24 hr after i.c.v. administration of 0, 1, 5, 10, 20 or 40 nmol of beta-FNA. The Kd and Bmax values of [3H]DAMGO binding were unaltered in animals treated with saline or 1 nmol of beta-FNA, whereas treatment with 5, 10 and 20 nmol of beta-FNA increased Kd and decreased Bmax values. The 40 nmol dose did not affect the Kd but decreased the Bmax value. Administration of 40 nmol of beta-FNA i.c.v. was not found to affect either the Kd or Bmax of delta opioid receptors assessed with [3H]DPDPE. Although some brain regions appeared to be affected to a greater degree than others, the autoradiographic localization of [3H]DAMGO binding at 10 different brain levels revealed a generally homogeneous loss of binding after 40 nmol of beta-FNA. beta-FNA appears to alkylate mu opiate receptors throughout the brain after i.c.v. administration. PMID- 8229782 TI - BAY X1005, a new inhibitor of leukotriene synthesis: in vivo inflammation pharmacology and pharmacokinetics. AB - (R)-2-[4-(quinolin-2-yl-methoxy)phenyl]-2-cyclopentyl acetic acid) (BAY X1005) is an orally active inhibitor of the synthesis of the leukotrienes B4 and C4 in selected animal models that effectively reduces the vascular phenomena of inflammation, i.e., edema formation and leukocyte immigration. The arachidonic acid-induced mouse ear inflammation test allowed the evaluation of the antiedematous effects of BAY X1005 after topical (ED50, 18 micrograms/ear) and oral (ED50, 48.7 mg/kg) administration. Profound inhibition of myeloperoxidase activity as a marker for phagocyte infiltration was seen (ED50, 3 micrograms/ear topically and 7.9 mg/kg p.o.) even 5 hr after application. The platelet activating factor-induced death of mice was statistical significantly and dose dependently reduced (100 mg/kg p.o.; mean, 51%). BAY X1005 had no analgesic properties in the phenyl-benzoquinone writhing test in mice and only limited efficacy in the baker's yeast-induced hyperalgesia test in the rat (ED50, 90 mg/kg p.o.), although cyclooxygenase inhibitors (indomethacin ED50, 1.7 mg/kg p.o.) are very potent. In another cyclooxygenase-sensitive test, the carrageenan induced edema and the baker's yeast-induced fever in the rat, BAY X1005 was virtually devoid of any activity. The rat whole blood ex vivo leukotriene B4 inhibition assay demonstrated that BAY X1005 was potent (ED50, 11.8 and 6.7 mg/kg p.o. at 1 and 5 hr, respectively) and had a long duration of action (16-hr ED40, 70 mg/kg p.o.). Similarly, inhibition of the zymosan-induced exudate leukotrienes B4 and C4 inhibition confirmed these data (ED50, 8.3 and 10.5 mg/kg p.o., respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229783 TI - Effects of aminohydroxybutane bisphosphonate on bone growth when administered after hind-limb bone loss in tail-suspended rats. AB - The effects of aminohydroxybutane bisphosphonate (AHBuBP) on bone after disuse osteopenia were studied in tail-suspended rats. Male Sprague-Dawley rats (weight range, 313-352 g) randomized into four groups of eight animals received 2 ml kg-1 day-1 of either AHBuBP (0.3 mg kg-1 day-1) or normal saline (vehicle) subcutaneously on days 14 and 15 of a 28-day experiment. The groups were 1) nonsuspended, saline; 2) suspended on days 14 to 28, saline; 3) suspended on days 0 to 28, AHBuBP; and 4) suspended on days 0 to 28, saline. On days 19 and 26, all rats received 15 mg/kg (1 ml/kg) of calcein. On day 28, they were sacrificed and their tibias and femurs were analyzed in vitro for bone density, strength and stiffness. The tibias were also analyzed histomorphometrically. The tibias and femurs from AHBuBP-treated rats were as dense as those in the nonsuspended group, whereas tail suspension in the untreated rats for 14 and 28 days caused a significant decrease in bone density. However, in measurements of bone strength and stiffness, the samples from the rats that received AHBuBP were similar to those of untreated rats suspended for 14 days, suggesting the newly formed bone was weaker. In the AHBuBP group, compared with all others, static histologic measurements of the proximal tibial metaphyses showed an increased bone area and perimeter and a decreased percentage of osteoid perimeter without a difference in the percentage of eroded perimeter. Dynamic histologic studies showed a decreased bone formation rate and decreased longitudinal growth rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229784 TI - Molecular basis for regionally specific action of ethanol on gamma-aminobutyric acidA receptors: generalization to other ligand-gated ion channels. AB - The present investigation provides evidence that there is neuroanatomical specificity for ethanol enhancement of gamma-aminobutyric acid (GABA)-induced inhibition in mammalian brain and that the expression of a specific GABAA isoreceptor is associated with this regional action of ethanol. Ethanol enhanced responses to iontophoretically applied GABA in the medial septum, inferior colliculus, substantia nigra reticulata, ventral pallidum and the diagonal band of Broca. In contrast to these results, responses to GABA applied to cells in the lateral septum, ventral tegmental area and the hippocampus were not affected by ethanol. In those brain regions where ethanol enhanced responses to GABA, a high concentration of zolpidem binding was found, whereas zolpidem binding was much lower or absent in brain regions where ethanol did not enhance GABA. These observations support the hypothesis that ethanol enhances GABA within specific regions of brain by affecting a GABAA receptor with specific structural components. From data obtained with in situ hybridization, there was a strong relationship between the regional distribution of zolpidem binding and the expression of specific mRNAs for the alpha-1, beta-2 and gamma-2 GABAA receptor subunits at sites where ethanol enhanced responses to GABA. The mRNA for the long and short variants of the gamma-2 subunit were found in brain regions both sensitive and insensitive to the action of ethanol on GABA-induced inhibition. These data were not able to address whether the gamma-2 long variant in combination with the alpha-1 and beta-2 subunits is essential for ethanol enhancement of responses to GABA. However, the observation that the long version of the gamma-2 subunit is present in brain areas where ethanol did not affect GABA function suggests that the presence of the long variant of the gamma-2 subunit alone is not sufficient for ethanol's action to enhance responses to GABA. Rather it is concluded that the appropriate combination of GABAA receptor subunits is critical for this action of ethanol. Because the GABAA receptor belongs to a superfamily of ligand-gated ion channels, the action of ethanol was examined on responses to agonists acting on receptors linked to other ion channels. As noted for GABA, local application of ethanol altered responses to NMDA, nicotine and glycine when applied to some, but not all, neurons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229785 TI - Duration and dose-related effects of an orally administered, partially lipophilic polyaminocarboxylic acid on the decorporation of plutonium and americium. AB - A recently developed, orally administered, partially lipophilic polyaminocarboxylic acid-based chelator, docosyl-triethylenetetraminepentaacetic acid (C22TT), was tested for its ability to promote decorporation of 239Pu and 241Am. The effects of dose and duration of treatment were determined in rats injected with 239Pu or 239Pu/241Am 2 weeks before the initiation of C22TT treatment and compared with untreated controls. In the dose-effects study, significant reductions in total body Am content were seen within 3 days after the initiation of C22TT treatment. After 30 days of treatment, there were dose related reductions in the Pu and Am content of soft tissues and bones. All doses of C22TT resulted in substantial reductions in Pu and Am content of the liver. In the time-response study, there were rapid reductions in total body Am content in the C22TT-treated animals. The greatest reductions occurred within the first 30 days of treatment. Significant decreases in Pu content of soft and hard tissue were observed in the treated animals at 30, 60 or 90 days compared with untreated controls. The greatest reductions in organ Pu content occurred within the first 30 days of treatment, particularly in the liver, but it continued throughout the experiment. Neutron-induced autoradiography showed that C22TT greatly reduced the incorporation of Pu into new bone and substantially reduced the Pu content of the bone marrow. There was no evidence of overt toxicity in either experiment. This study demonstrates that orally administered C22TT is effective in reducing soft and hard tissue content of internally deposited Pu and Am. PMID- 8229786 TI - Cerebral flavin-containing monooxygenase-mediated metabolism of antidepressants in brain: immunochemical properties and immunocytochemical localization. AB - Flavin-containing monooxygenase (FMO)-mediated oxidation of the model substrates N,N-dimethylaniline and methimazole, and the antidepressants imipramine and fluoxetine, was determined in rat brain microsomes. No sex-related difference was observed in the activity of the FMO-mediated metabolism of the four substrates examined. The Km values for flavin-containing monooxygenase-mediated metabolism of N,N-dimethylaniline and methimazole were 2.8 and 0.8 mM, respectively, and the Km values for the oxidation of the antidepressants imipramine and fluoxetine were 20.9 and 9.8 microM, respectively. The Vmax values for oxidation of N,N dimethylaniline, methimazole, imipramine and fluoxetine were 340, 31, 182 and 470 nmol nicotinamide adenine dinucleotide phosphate oxidized/min/mg protein, respectively. Western immunoblot analysis using antisera to purified porcine liver FMO did not reveal any immunological cross-reactivity with male or female brain microsomal protein. Antibody to rabbit lung flavin-containing monooxygenase cross-reacted with brain microsomes as examined by Western immunoblot studies. Addition of the antibody raised against rabbit lung FMO resulted in inhibition (43% inhibition) of the FMO-mediated metabolism of imipramine. Immunocytochemical examination of rat brain sections using the above antibody revealed the preferential localization of flavin-containing monooxygenase in the neuronal cell body. The flavin-containing monooxygenase-mediated metabolism of antidepressant drugs by brain microsomes is of profound pharmacological significance. PMID- 8229787 TI - Ethanol consumption by the nursing mother induces cytochrome P-4502E1 in neonatal rat liver. AB - Cytochrome P-4502E1 (P-4502E1) is not present in fetal rat liver because activation of the gene occurs shortly after birth. Ethanol is an inducer of P 4502E1 in adult rats. Studies were carried out to evaluate whether transplacental induction of P-4502E1 by ethanol can occur after oral consumption of ethanol by the pregnant mother. Because ethanol can be excreted in breast milk, the possible induction of P-4502E1 in neonatal liver when ethanol was consumed during the gestational and neonatal period by the mother was also determined. Pregnant rats received control or an ethanol-containing liquid diet starting on the 9th day of gestation and were killed on the 17th day or 21st day, of gestation or allowed to deliver. The rats continued on their respective diets for the first 2 weeks of the neonatal period. P-4502E1 messenger RNA (mRNA), protein or catalytic activity was not detectable in fetal liver and was not induced in the fetuses from the ethanol-consuming mothers. Transplacental induction of P-4502E1 by ethanol did not occur in this model. Induction by ethanol of P-4502E1 protein and catalytic activity but not mRNA occurred in maternal liver. P-4502E1 mRNA, protein and catalytic activity were detected shortly after birth and increased over the 2 week neonatal period. The P-4502E1 content and oxidation of p-nitrophenol or dimethylnitrosamine by hepatic microsomes from neonates of mothers consuming the ethanol diet were increased 2- to 3-fold compared with controls however, P-4502E1 mRNA levels were not elevated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229788 TI - Preclinical studies on LY228729: a potent and selective serotonin1A agonist. AB - LY228729 is a conformationally restricted tryptamine derivative with a carboxamide serving as a protophilic group to mimic the hydroxyl in serotonin (5 HT). LY228729 has high affinity for the 5-HT1A receptor, weak affinity for the 5 HT1D receptor and no significant affinity for other monoaminergic receptors studied. LY228729 was less effective than 5-carboxamidotrytamine in suppressing K(+)-evoked release of 3H-5-HT from parietal-occipital cortical slices from guinea pigs, which is in agreement with its weak 5-HT1D receptor affinity. LY228729 reduced hypothalamic 5-hydroxyindole-3-acetic acid levels and increased serum corticosterone levels in rats. LY228729 reduced hypothalamic 5 hydroxytryptophan accumulation after decarboxylase inhibition. LY228729 increased flat posture and lower lip retraction scores in rats at doses between 0.1 and 1 mg/kg s.c. (p.o. doses were 10 times higher) and these effects were blocked by (+/-) pindolol. LY228729 induced a hypothermic response in rats, which was blocked by (+/-) pindolol. These in vivo responses are characteristics of compounds with 5-HT1A agonist activity. In the preclinical efficacy models, LY228729 suppressed motion sickness responses in cats; decreased ejaculatory latency and the increased copulatory efficiency and rate in rats and increased punished responding at lower doses than it lowered unpunished responding in rats. Collectively, these results indicate that LY228729 is potent 5-HT1A agonist with bioavailability properties sufficient for clinical evaluation and with efficacy in preclinical models of anxiety, sexual disorders and motion sickness. Since the 5-HT1A agonists that have been studied previously have antidepressant activity, this indication will also be evaluated. PMID- 8229789 TI - Potassium channel blockade and halothane vasodilation in conducting and resistance coronary arteries. AB - On the basis of reports that volatile anesthetics, such as halothane, open membrane potassium channels in several tissues, it was investigated whether coronary vasodilation by halothane is mediated by a similar mechanism. The ability of glyburide, a blocker of ATP-sensitive K+ (KATP) channels, and tetraethylammonium (TEA+), a blocker of Ca(2+)-activated K+ channels, to modify halothane-induced vasodilation was assessed in two vascular preparations. First, coronary resistance vessel tone was measured in isolated rat hearts arrested with tetrodotoxin and, second, conducting vessel responsiveness was evaluated in ring segments of the porcine epicardial coronary artery contracted with prostaglandin F2 alpha. Halothane alone markedly vasodilated the perfused hearts and attenuated the agonist contraction of the coronary rings. Blockade of KATP channels with glyburide alone did not affect the base-line vascular tone or responsiveness but it inhibited cromakalim vasodilation. TEA+ alone caused vasoconstriction. In hearts perfused at constant pressure, glyburide significantly attenuated the halothane-induced increase in coronary flow by 56% and perfusion with a high K+ buffer reduced the halothane-induced vasodilation response by 94%. In endothelium denuded coronary rings, glyburide did not affect halothane-induced relaxation but KATP channel blockade potentiated halothane-caused relaxation in endothelium intact rings. The attenuation of halothane-induced vasodilation by TEA+ seen in the perfused hearts did not achieve statistical significance and no halothane/TEA+ interaction was evident in the coronary rings. Thus, the data from the perfused heart experiments suggest that halothane relaxes rat coronary resistance vessels, in part, by opening K+ channels. PMID- 8229790 TI - Prolactin increases Na+/taurocholate cotransport in isolated hepatocytes from postpartum rats and ovariectomized rats. AB - The role of prolactin (PRL) in regulating the transport of the bile acid taurocholate (TC) was assessed using isolated rat hepatocytes. Na(+)-dependent TC cotransport was determined in hepatocytes from female nonpregnant, pregnant (19 20 days pregnant), postpartum (48 hr postpartum) and postpartum rats treated with bromocriptine to block PRL secretion. In separate experiments ovariectomized rats were infused i.v. with solvent alone (OVX) or with ovine PRL (100, 300 and 600 micrograms/day) for 7 days (OVX+oPRL). The least squares estimates of Km (microM) and Vmax (nmol/min/mg protein) for Na(+)-dependent TC uptake were, respectively: 15 and 1 in nonpregnant, 9 and 0.4 in pregnant, 9 and 1.1 in postpartum and 15 and 1 in bromocriptine-treated postpartum rats, and were 15 and 1 in OVX, 15 and 1 in OVX+oPRL (100 micrograms/day), 30 and 2 in OVX+oPRL (300 micrograms/day) and 18 and 2 in OVX+oPRL (600 micrograms/day) rats, respectively. Calculation of the 95% joint confidence limits for Km and Vmax showed that Na(+)-dependent TC uptake was significantly decreased in pregnant rats, and significantly increased in postpartum rats relative to nonpregnant controls. Bromocriptine-treated postpartum rats were not different from controls. Infusion of 300 and 600 micrograms/day oPRL significantly increased Na(+)-dependent TC transport relative to OVX rats. Na(+)-K(+)-ATPase activity did not differ among the groups. These data indicate that PRL is responsible for the increased Na(+)-dependent transport of TC in the maternal liver postpartum, and that administration of oPRL to ovariectomized rats increases this transport in a dose-dependent manner. PMID- 8229791 TI - Neuropeptide FF receptors: structure-activity relationship and effect of morphine. AB - Neuropeptide FF (FLFQPQRFamide, NPFF) is an octapeptide implicated in morphine analgesia, tolerance and dependence. Many of the behavioral effects of NPFF have also been observed with the invertebrate neuropeptide Phe-Met-Arg-Phe-amide (FMRFamide), which binds to NPFF receptors because of its low homology to the C terminal portion of NPFF. A competitive ligand binding assay was used to characterize NPFF receptors in rat spinal cord and a strong requirement was found for the C-terminal Arg-Phe-amide. It was found that FMRFamide (Ki = 1.8 nM) bound with lower affinity than NPFF (0.26 nM) but it was about 7-fold more potent than PQRFamide (12 nM). This finding explains the similar bioactivities of NPFF and FMRFamide. The Gln2 appeared to be the cause of the relatively low potency of PQRFamide, based on the binding specificity of NPFF receptors for a series of FMRFamide analogs. In contrast to the Arg-Phe-amide, substitutions at the first and second positions of FMRFamide were generally tolerated, with the most potent analogs being PMRFamide (Ki = 0.54 nM), FFRFamide (0.25 nM) and FWRFamide (0.42 nM). Among the most potent ligands was a pentapeptide containing a photoreactive Phe analog, D-Tyr-(p-benzoyl-Phe)-norLeu-Arg-Phe-amide (Ki = 0.23 nM). It was found that dansyl-PQRFamide and dansyl-RFamide also bound to NPFF receptors with Ki values of 6.1 and 73 nM, respectively. The radioligand binding and G-protein coupling of NPFF receptors were not altered by chronic morphine treatment. PMID- 8229792 TI - Pharmacological examination of receptors mediating contractile responses to tachykinins in airways isolated from human, guinea pig and hamster. AB - The abilities of agonists selective for neurokinin (NK)-1 (Ac [Arg6,Sar9,Met(O2)11]-SP6-11, ASMSP), NK-2 ([beta-Ala8]-NKA4-10) and NK-3 ([Asp5,6,MePhe8]-SP5-11, senktide analog) receptors to contract human bronchus and guinea pig and hamster trachea were studied. The antagonism of these responses by selective antagonists was also examined. In the human bronchus and hamster trachea, [beta-Ala8]-NKA4-10 was the most potent agonist, whereas ASMSP and senktide analog failed to elicit contractions greater than 50% of the maximum response even at concentrations reaching 1 to 3 x 10(-4) M. By contrast, both ASMSP and [beta-Ala8]-NKA4-10 were potent contractile agonists in guinea pig trachea. In all tissues, the selective NK-1 receptor antagonist (2S,3S)-cis-2 (diphenylmethyl)-N-[(2-methoxyphenyl)-methyl]-1-a zab icyclo- [2.2.2]octan-3 amine (CP 96,345) was without effect on contractile responses to [beta-Ala8]-NKA4 10. Blockade by CP 96,345 of responses to ASMSP was, however, observed in the guinea pig trachea, but not in human bronchus or hamster trachea. Responses to ASMSP in human bronchus and hamster trachea were inhibited by NK-2 antagonists, whereas these compounds had little effect on responses to ASMSP in guinea pig trachea. In all tissue types, responses to senktide analog were inhibited by NK-2 antagonists.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229793 TI - Thapsigargin-evoked changes in human platelet Ca2+, Na+, pH and membrane potential. AB - 1. In this work we explored the effect of thapsigarin on the intracellular Ca2+, pH, Na+ and membrane potential in human platelets. These parameters were monitored using the fluorescent probes fura-2, 2',7'-bis-(2-carboxyethyl)-5,6 carboxyfluorescein, sodium-binding benzofuran isophthalate, and 3,3' dipropylthiadicarbocyanine iodide. 2. Thapsigargin caused an increase in the cytosolic Ca2+, coupled with cytosolic alkalinization. Thapsigargin-induced alkalinization was Na(+)-dependent, indicating that thapsigargin stimulated the Na(+)-H+ exchange. 3. Using Mn2+ as a Ca2+ surrogate, we showed that thapsigargin activated Ca2+ channels at relatively low levels of cytosolic Ca2+, suggesting that a rise in cytosolic free Ca2+ is not the signal for the activation of these channels. 4. Thapsigargin-induced increase in the cytosolic free Ca2+ was greater in Na(+)-containing medium than in Na(+)-free medium, suggesting that Na(+) dependent mechanisms participate in the regulation of platelet cytosolic Ca2+. 5. Thapsigargin not only increased the cytosolic Ca2+, but also elevated the cytosolic free Na+. The latter effect was more pronounced in Ca(2+)-free medium, a finding that may indicate that some of the Na+ enters through Ca2+ entry pathways. 6. Finally, thapsigargin evoked sustained platelet hyperpolarization which was attenuated by charybdotoxin, indicating thapsigargin-induced stimulation of Ca(2+)-sensitive K+ channels. 7. Together these observations demonstrate a multifactorial effect of thapsigargin on platelets that can be utilized to further understand platelet ionic homeostasis. PMID- 8229794 TI - The role of the kidney in canine blood pressure control: direct assessment of the closed-loop gain. AB - 1. The feedback control of arterial blood pressure by the kidney in the range of hours was investigated in resting, conscious foxhounds. 2. A servo-control device (connected to an aortic occlusive cuff implanted above both renal arteries) was used to maintain a constant pressure difference of 20 mmHg between aortic pressure measured proximal (mean arterial blood pressure: MAP) and distal (renal artery pressure: RAP) to the aortic cuff. 3. Protocol 1 (n = 6) served as a 4 h time control without intervention, protocol 2 (n = 6) consisted of three periods: after a control of 20 min duration, the servo-control device was activated for 180 min; this was followed by a recovery period of 40 min. Protocol 3 (n = 6) was as protocol 2, but during converting-enzyme inhibition. 4. Servo-control increased plasma renin activity (PRA) transiently from 0.5 ng angiotensin I (AI) ml-1 h-1 to a peak value of 2.4 ng AI ml-1 h-1, subsequently both RAP and MAP rose to reach a new steady state. During this increase in RAP, PRA declined to 1.4 ng AI ml-1 h-1. 5. On average, the compensation of the pressure decrease sensed by the kidney amounted to 63% of the error signal (closed-loop gain of 0.63 +/- 0.1). 6. Converting-enzyme inhibition reduced this closed-loop gain significantly (protocol 2 vs. protocol 3, 0.63 +/- 0.1 vs. 0.15 +/- 0.1; P < 0.05). 7. It is concluded, that the kidney plays an important role in medium-term blood pressure regulation, most probably via the renin-angiotensin system. PMID- 8229795 TI - Multiple effects of spermine on N-methyl-D-aspartic acid receptor responses of rat cultured hippocampal neurones. AB - 1. The modulation by polyamines of responses to N-methyl-D-aspartic acid (NMDA) was studied using a rapid perfusion system and whole-cell voltage-clamp recording from rat hippocampal neurons in dissociated culture. 2. Concentration jump responses to 100 microM NMDA in the presence of 10 microM glycine revealed potentiation by 3 mM spermine at a membrane potential of +60 mV, but depression at -120 mV; the degree of potentiation at +60 mV was variable from cell to cell while marked depression at -120 mV was observed in all cells. The depression of responses to NMDA by spermine was highly voltage dependent (z delta = 1.17) with an apparent equilibrium dissociation constant for block at 0 mV of 27 mM. 3. Analysis of spermine dose-potentiation curves for responses recorded at +60 mV in the presence of 10 microM glycine revealed a half-maximal effect at 125 microM. Under the same conditions, but at -60 mV, analysis of spermine-evoked depression was performed for cells with less than 5% potentiation at +60 mV, and revealed half-maximal inhibition at 344 microM. 4. Dose-response analysis for the glycine sensitive activation of NMDA receptors at +60 mV revealed a 3.5-fold increase in apparent affinity for glycine in the presence of 1 mM spermine. This increase in affinity for glycine was accompanied by a 3.3-fold decrease in the rate of development of glycine-sensitive desensitization, and a 2.4-fold decrease in the rate of dissociation of glycine from NMDA receptors, while the rate constant for dissociation of NMDA was not reduced. 5. In the presence of non-saturating concentrations of glycine, spermine-induced potentiation at +60 mV developed with two exponential components: a slow glycine-sensitive component, the amplitude and time constant of which decreased with increasing glycine concentration (30 nM glycine, amplitude = 80.2 +/- 5.1%, tau = 780 +/- 79 ms; 3 microM glycine, amplitude = 22.6 +/- 7.1%, tau = 45 +/- 13 ms), and a faster component (tau < 20 ms at all concentrations of glycine), the amplitude of which varied from cell to cell, and which became larger with increase in concentration of glycine. When responses to the application of spermine were measured in the presence 10 microM L-alanine instead of 100 nM glycine, the slow component of potentiation was absent.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229796 TI - Time course of spontaneous calcium-activated chloride currents in smooth muscle cells from the rabbit portal vein. AB - 1. The time course of spontaneous calcium-activated chloride currents was studied with the perforated patch technique in freshly dispersed smooth muscle cells from the rabbit portal vein. 2. In potassium-containing solutions the spontaneous transient outward current (STOC, a calcium-activated potassium current) was more commonly recorded than spontaneous transient inward currents (STICs, a calcium activated chloride current). In addition the duration of STOCs was much briefer (about 100 ms) than the duration of STICs (about 400 ms). 3. The decay of STICs could be described by a single exponential but the STOC decay appeared to be more complex. The decay time constant of STICs was not determined significantly by amplitude. 4. The time constant of decay of STICs (tau) was 86 ms at -50 mV and was increased by depolarization. Between -90 and +50 mV the relationship between tau and membrane potential was exponential and tau changed e-fold for a change of membrane potential of 120 mV. 5. The I-V relationship of STIC amplitude was linear between -10 and +50 mV but at more negative potentials the chord conductance was reduced and the I-V relationship exhibited negative slope conductance between -50 and -90 mV. 6. There was good agreement between the STIC tau values and the exponential relaxations to voltage steps evoked during caffeine-induced calcium-activated chloride currents. 7. In the presence of the chloride channel blocking agent anthracene-9-carboxylic acid the STIC amplitude was reduced and tau was increased. This effect was voltage dependent with a much greater effect at positive potentials. 8. The evidence suggests that the decay of STICs represents closure of chloride channels and tau approximates to the channel mean open time. PMID- 8229797 TI - Regulation of the intracellular free calcium concentration in acutely dissociated neurones from rat nucleus basalis. AB - 1. Neurones were acutely dissociated from the rat nucleus basalis. Whole-cell patch clamp recordings of calcium currents (ICa) and fura-2 microfluorimetric recordings of intracellular free Ca2+ concentration ([Ca2+]i) were made simultaneously. 2. Depolarization from -60 to 0 mV elicited ICa and a gradual increase in [Ca2+]i. After repolarization, ICa terminated in 0.7 ms, and [Ca2+]i recovered to control exponentially (1-5 s). 3. Both ICa and the transient [Ca2+]i increase in response to step depolarizations, were abolished in Ca2+ free extracellular solution and in Cd(2+)-containing solution. 4. Depolarizations from -90 mV to membrane potentials less negative than -40 mV induced ICa and an increase in [Ca2+]i. Depolarization to 0 mV elicited the maximum ICa, and produced the largest increase in [Ca2+]i. There was a parallel relationship between the [Ca2+]i increase and the magnitude of the ICa. 5. The [Ca2+]i increase was associated with an increase in total Ca2+ influx when the duration of the step depolarization was varied. The relationship between the total Ca2+ influx and the peak of [Ca2+]i transient reached an asymptote as total Ca2+ influx exceeded 200 pC. A similar finding was made when more than thirty action potentials were used in increasing [Ca2+]i. 6. The process of the [Ca2+]i recovery was slowed down by lowering the temperature, by an intracellular dialysis with vanadate, by extracellular application of a mitochondrial inhibitor, carbonyl cyanide m-chlorophenyl-hydrazone (CCCP), and by Na(+)-free external solution. It was unaffected by membrane potential (-50 to -130 mV). 7. When pipette solution contained a high concentration of fura-2 (200 microM), the [Ca2+]i increase per 1 pC of Ca2+ influx decreased, and the [Ca2+]i recovery was slowed. 8. The results indicate that the ICa through voltage-dependent Ca2+ channels elevates [Ca2+]i. The neurones possess a large capacity for Ca2+ buffering, and the recovery of [Ca2+]i requires both the Ca2+ pump and membrane Na(+)-Ca2+ exchange. PMID- 8229798 TI - Muscle stiffness, strength loss, swelling and soreness following exercise-induced injury in humans. AB - 1. In order to study injury-related changes in muscle stiffness, injury to the elbow flexors of thirteen human subjects was induced by a regimen of eccentric exercise. 2. Passive stiffness over an intermediate range of elbow angles was measured with a device which held the relaxed arm of the subject in the horizontal plane and stepped it through the range of elbow angles from 90 deg to near full extension at 180 deg. The relation between static torque and elbow angle was quite linear over the first 50 deg and was taken as stiffness. 3. Stiffness over this range of angles more than doubled immediately after exercise and remained elevated for about 4 days, and may result from low level myofibrillar activation induced by muscle stretch. 4. Arm swelling was biphasic; arm circumference increased by about 3% immediately after exercise, fell back toward normal, then increased by as much as 9% and remained elevated for as long as 9 days. 5. Ultrasound imaging showed most of the swelling immediately following the exercise to be localized to the flexor muscle compartment; subsequent swelling involved other tissue compartments as well. 6. Muscle strength declined by almost 40% after the exercise and recovery was only slight 10 days later; the half-time of recovery appeared to be as long as 5-6 weeks. PMID- 8229799 TI - Membrane properties of rat suprachiasmatic nucleus neurons receiving optic nerve input. AB - 1. The electrophysiological properties of suprachiasmatic nucleus (SCN) neurons (n = 33) receiving optic nerve input were studied with intracellular recordings in rat hypothalamic slices maintained in vitro. Our major goal was to provide baseline data concerning the intrinsic membrane properties of these neurons and to test the hypothesis that the neurons are homogeneous electrophysiologically. 2. Action potentials were short in duration and followed by a pronounced hyperpolarizing after-potential. Spike amplitude (58.2 +/- 1.1 mV, mean +/- S.E.M.; measured from threshold), spike duration (0.83 +/- 0.03 ms; measured at half amplitude) and hyperpolarizing after-potential amplitude (23.9 +/- 1.0 mV; measured from threshold) appeared unimodally distributed and did not co-vary. 3. Intracellular injection of depolarizing current pulses evoked spike trains, and spike inactivation, spike broadening and frequency accommodation were always present. An after-hyperpolarization followed the spike train in all but one neuron. 4. Membrane time constant ranged from 7.5 to 21.1 ms (11.4 +/- 0.7 ms, n = 27), and its distribution appeared to be unimodal with the peak at approximately 10 ms. Input resistance ranged from 105 to 626 M omega (301 +/- 23 M omega, n = 33); the distribution also appeared unimodal with its peak at approximately 250 M omega. 5. A subpopulation (16 of 33, 48%) of the neurons exhibited slight (6-29%) time-dependent inward rectification in their voltage responses to hyperpolarizing current injection. Of the neurons lacking the time dependent rectification, some (n = 5) exhibited time-independent inward rectification of 6-20% and others showed no (or < 3%) such rectification. The degree of inward rectification was correlated with neuronal excitability (r = 0.60, P < 0.002; assessed by measuring the steepness of the primary slope of the frequency-current plot) and with the spontaneous firing rate (r = 0.49, P < 0.007). Furthermore, the neurons with > 6% inward rectification (neurons with time-dependent and -independent rectification were combined) were more excitable (362 +/- 43 Hz/nA (n = 15) vs. 221 +/- 37 Hz/nA (n = 9), P < 0.05) and had a higher spontaneous firing rate (11.1 +/- 1.9 Hz (n = 19) vs. 3.9 +/- 1.5 Hz (n = 11), P < 0.02) than the neurons with no or negligible (i.e. < 3%) inward rectification. The two groups, however, were not significantly different in membrane time constant and input resistance. 6. When adequately hyperpolarized, twelve of seventeen (71%) neurons generated small low-threshold spike (LTS) potentials in response to depolarizing current pulses.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229800 TI - Characteristics of Ca2+ release induced by Ca2+ influx in cultured bullfrog sympathetic neurones. AB - 1. A rise in intracellular Ca2+ ([Ca2+]i) and a Ca2+ current (ICa) induced by a depolarizing pulse were simultaneously recorded by fura-2 or indo-1 fluorescence and whole-cell patch clamp techniques in cultured bullfrog sympathetic ganglion cells. 2. [Ca2+]i (calculated from the ratio of fura-2 fluorescences excited at 380 and 340 nm and recorded with a photomultiplier at > 492 nm) rose regeneratively (in most cells) during a command pulse (from -60 to 0 mV, 100 ms), continued to rise thereafter, peaked at 666 ms (on average) and decayed slowly with a half-decay time of 22.8 s. 3. Scanning a single horizontal line across the cytoplasm with an ultraviolet argon ion laser (351 nm) and recording indo-1 fluorescences at two wavelengths (peaked at 410 and 475 nm) with a confocal microscope demonstrated that [Ca2+]i beneath the cell membrane rose much faster than that in the deeper cytoplasm. The time course of the spatial integral of [Ca2+]i, however, corresponded well with that recorded with fura-2 fluorescence using a photomultiplier. 4. [Ca2+]i measured by fura-2 fluorescence ratio using a photomultiplier did not increase during a strong depolarizing pulse (-60 to +80 mV), but sometimes rose after the pulse. A depolarization-induced rise in [Ca2+]i ([Ca2+]i transient) was blocked in a Ca(2+)-free, EGTA solution, reduced by lowering the extracellular Ca2+ concentration ([Ca2+]o) to 0.45 or 0.9 mM and enhanced by raising [Ca2+]o to 7.2 or 14.4 nM. 5. The extracellular Ca2+ dependence was non-linear when long depolarizing pulses (up to 500 ms) were applied; the amplitude of [Ca2+]i transient/Ca2+ entry (unit [Ca2+]i transient) increased with an increase in Ca2+ entry. 6. Increasing the duration of depolarization (-50 or -60 to 0 mV) from 20 to 500 ms enhanced asymptotically the integral of ICa (due to inactivation), and progressively the magnitude of [Ca2+]i transients, leading to the apparent non-linear dependence of unit [Ca2+]i transient on Ca2+ entry as well as on the duration of membrane depolarization. The peak time of [Ca2+]i transient was unchanged for pulse durations up to 300 ms, but prolonged with an increase in pulse duration to 500 ms. 7. Inhibitors of Ca2+ release from intracellular Ca2+ reservoirs, dantrolene (10 microM) and ryanodine (50 microM), blocked the [Ca2+]i transient to 56 and 30%, respectively, of the control. 8. The higher the basal [Ca2+]i level, the greater was the magnitude of the [Ca2+]i transients.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229801 TI - The role of bicarbonate in GABAA receptor-mediated IPSPs of rat neocortical neurones. AB - 1. The ionic mechanism underlying the fast, GABAA receptor-mediated inhibitory postsynaptic potential (IPSPA) was examined in rat neocortical neurones using intracellular recording techniques. Synaptic responses were evoked by orthodromic stimulation applied to the subcortical white matter or to the pial surface. All experiments were carried out at a constant extracellular Cl- concentration. 2. The resting membrane potential was -76.2 +/- 1.0 mV (mean +/- S.E.M., n = 32) and in most cells IPSPA was depolarizing. The reversal potential of IPSPA (EIPSP-A) was -70.2 +/- 0.9 mV (n = 32) and that of a more slowly developing hyperpolarizing response (IPSPB) was -91.4 +/- 1.3 mV (n = 28). 3. An examination of the temporal relationships between excitatory postsynaptic potentials (EPSPs) and IPSPAs in different cells suggested that, despite partial overlap of these responses, EPSPs had little influence on the measured values of EIPSP-A. 4. Application of 20 mM trimethylamine (TriMA), a membrane-permeant weak base which is expected to produce a rise in pHi (and hence in intracellular HCO3-), induced a reversible positive shift in EIPSP-A of up to +9.0 mV (mean + 4.2 mV) at an extracellular pH (pHo) of 7.4. In some experiments, the shift in reversal potential was associated with a change in the polarity of IPSPA from hyperpolarizing to depolarizing. 5. Application of 20 mM lactate (a membrane permeant weak acid which is expected to produce a fall in pHi and hence in intracellular HCO3-) at pHo 7.0 produced a hyperpolarizing shift in EIPS-A of up to -7.5 mV (mean -5.6 mV). In some experiments, exposure to lactate changed the polarity of IPSPA from depolarizing to hyperpolarizing. 6. Changes in pHo from 7.4 to 7.0 reduced the effect of TriMA and augmented that of lactate on EIPSP-A, as could be expected on the basis of the pHo-dependent change in the fraction of membrane permeable non-charged weak base or acid. 7. Under control conditions, a change in pHo from 7.4 to 7.0 produced a slight positive shift (< +2 mV) in EIPSP A. In the presence of TriMA, a similar change in pHo gave rise to a negative shift (-1.8 to -2.7 mV). 8. The results obtained indicate that HCO3- ions contribute significantly to the IPSPA, thereby making EIPSP-A more positive than the Cl- equilibrium potential.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229802 TI - Decreased energy requirement of toad retina during light adaptation as demonstrated by 31P nuclear magnetic resonance. AB - 1. The effect of light and dark adaptation on the levels of phosphorus metabolites (nucleotide di- and triphosphates, phosphocreatine, pyridine nucleotide, inorganic phosphate, phosphodiesters, phosphomonoesters, and uridine diphosphate-glucose) in the toad (Bufo marinus) retina and retinal extracts was studied by 31P nuclear magnetic resonance (NMR) spectroscopy. 2. Spectra were acquired using an NMR probe specifically designed for superfusion and illumination of a single retina. Retinae were maintained at a steady state for up to 10 h in an electrolyte solution containing 10 mM Hepes buffer and bubbled with 98% O2-2% CO2, pH 7.8 at 20 degrees C. 3. The intracellular concentrations of the phosphorus metabolites were measured in total darkness or during prolonged exposure to light. The concentration of nucleoside triphosphates (NTP) in the dark-adapted retina was about 1.5 mM and that of phosphocreatine (PCr) was about 0.7 mM. 4. In saturating levels of light, 6.0 x 10(11) or 1.5 x 10(13) quanta s-1 cm-2 at 520 nm, the levels of PCr and phosphomonoesters rose, the levels of NTP and protons (pH) were maintained, and the levels of pyridine nucleotides and nucleotide diphosphates (NDP) fell. 5. A rise in the level of PCr in the presence of an unchanged level of NTP in the light-adapted retina indicates that the energy consumption of the retina is greater in the dark. 6. These results are in agreement with the results of oxygen consumption, glucose dependence, and electrophysiological studies which also indicate that the metabolic energy requirement of the retina decreases in light. PMID- 8229803 TI - Single-channel study of the cyclic AMP-regulated chloride current in guinea-pig ventricular myocytes. AB - 1. Properties of the cyclic AMP-regulated Cl- channel were studied in guinea-pig ventricular myocytes with the patch clamp technique. Cell-attached patch recordings were performed, while the cell was dialysed with a cyclic AMP (0.2-0.5 mM)-containing internal solution through a second patch pipette. The latter pipette was also used to monitor the whole-cell Cl- conductance. 2. The whole cell showed a large Cl- conductance for 10-15 min after the beginning of cell dialysis. The activity of single Cl- channels began to appear in some of the cell attached patches during this time. 3. The channels showed a high open probability (0.69 +/- 0.14, mean +/- S.D., n = 12) at the time of their appearance, and the open probability did not appreciably increase thereafter, even when the whole cell Cl- conductance increased further with time. 4. An increase in the number of active channels was observed in some patches with progression of the cell dialysis. In such cases, the newly activated channels also showed a high open probability. 5. The above results are consistent with the hypothesis that the cyclic AMP system makes the 'latent' Cl- channels available without influencing their own kinetic behaviour. The available channels may intrinsically exhibit a high open probability. 6. Chloride channel currents could also be recorded in the outside-out patches excised from the cyclic AMP-loaded cells. The I-V relation of these currents showed outward rectification under the condition of symmetrical Cl gradients, suggesting that the channel itself or a related structure has the property of rectifying current flow. 7. The channel seemed to have at least one open state and two closed states; the open-time histograms showed one exponential component with the values of time constant scattering around 1 s, while the closed-time histograms showed two exponential components with the values of time constant scattering around 0.2 and 1 s. These time constants showed no clear voltage dependence. PMID- 8229804 TI - Two types of 4-aminopyridine-sensitive potassium current in rabbit Schwann cells. AB - 1. Delayed rectifier K+ currents were studied in Schwann cells cultured from neonatal rabbit sciatic nerves with the whole-cell patch-clamp technique. 2. Depolarizing voltage steps (40 ms duration) activated two types of K+ current: type I, whose apparent activation threshold was about -60 mV (half-maximal conductance at -40 +/- 1 mV, n = 10); and type II, whose apparent activation threshold was about -25 mV (half-maximal conductance at + 11 +/- 1 mV, n = 9). 3. Type I current was blocked by alpha-dendrotoxin (alpha-DTX) with an apparent equilibrium dissociation constant (KD) of 1.3 nM, whereas the type II current was unaffected by exposure to 500 nM toxin. The action of alpha-DTX on the type I current was reversible. 4. Most cells exhibited both types of current, but occasionally some cells displayed just type I or just type II. 5. Type I current activated rapidly and then showed a much slower fade, which became more noticeable with larger depolarizations. Activation of type II current was slower than that of type I and depended less steeply on voltage. The time constants of activation for type I and type II currents were derived with a Hodgkin-Huxley formalism (based on second-power activation and deactivation kinetics). The longest activation time constant for type II gating was more than twice the corresponding time constant for type I; however, the time constants determined from tail current decays at potentials more negative than -60 mV were shorter for the type II currents than for the type I currents. 6. Both type I and type II currents were sensitive to micromolar concentrations of 4-aminopyridine (4-AP). The KD for 4-AP blockade of type II current was 630 microM (pH 7.2, membrane potential (Em) = -10 mV), which is about 6 times higher than the corresponding value for 4-AP blockade of type I current at negative membrane potentials. The differential sensitivity of the type I and type II currents to 4-AP may account for the apparent voltage dependence of 4-AP block of delayed rectifier K+ currents. 7. In addition to types I and II, a third type of outward K+ current (type III) was generated in most cells at positive membrane potentials. This latter current was insensitive to millimolar concentrations of 4-AP. 8. Similarities between Schwann cell and neuronal potassium channels are discussed. PMID- 8229805 TI - Ionic dependence of a slow inward tail current in rat dorsal raphe neurones. AB - 1. The mechanism underlying a large slow inward tail current was studied in serotonergic dorsal raphe (DR) neurones. The tail current is most easily observed under conditions of suppressed K+ channel outward currents and follows the activation of a calcium current. This current may underlie a slow after depolarizing potential (ADP) which follows action potentials observed in acutely isolated DR neurones. 2. The after-hyperpolarizing potential (AHP) following action potentials which should reverse at EK (the reversal potential for potassium) becomes an ADP at less negative potentials than expected due to contamination by the slow inward tail current. 3. DR neurones were acutely isolated enzymatically; the ADP in current clamp and the tail current underlying it in voltage clamp were studied using the patch clamp method. When the external Na+ was replaced with TEA or choline the slow inward tail current was completely abolished. Blocking K+ channels from the inside of the cell membrane with 40 mM TEACl or large concentrations of internal Cs+ also blocked the slow inward tail current. 4. The tail current proved to be independent of calcium influx or intracellular calcium release as it was not affected by inorganic calcium channel blockers or caffeine. 5. The tail grew exponentially upon lengthening the depolarizing test pulse and appeared to reverse close to 0 mV indicating that the current was carried by a nonselective cation conductance. Removal of external Na+ and replacement with Li+ ions reversibly blocked the tail current by 77%. 6. The data rule out several mechanisms for the generation of the current, namely: a calcium-activated chloride conductance, a calcium-activated non-selective cation conductance, a Na(+)-Ca2+ exchange pump current or a sodium-activated K+ conductance. 7. The slow tail current may be explained by postulating an inward movement of Na+ through a channel which is blocked by high concentrations of external TEA and Li+ or internal Cs+ or 40 mM TEA. PMID- 8229806 TI - Effects of nerve growth factor on neuronal plasticity of the kitten visual cortex. AB - 1. The effect of intraventricular administration of nerve growth factor (NGF) by means of a cannula-minipump system was studied in kittens monocularly deprived during the critical period. The ocular dominance of area 17 neurones of NGF treated and control kittens was determined by conventional extracellular recordings. The soma size of cells in A and A1 laminae of the lateral geniculate nucleus (LGN) was also evaluated in Cresyl Violet preparations. 2. Binocularly responsive neurons were found to be significantly more numerous in NGF-treated than in control kittens. The shrinkage of cells from the deprived LGN laminae normally observed in control kittens was prevented by NGF administration. 3. Following an initial period of monocular deprivation (MD) kittens subsequently treated with NGF showed a substantial recovery of functional binocular connections. 4. These findings indicate that the administration of NGF during the period of deprivation reduces the amblyopic effects of MD, while its administration to kittens with both eyes open following the initial deprivation promotes recovery of the deprived eye. 5. Neurotrophic factors may contribute to the regulation of experience-dependent modifications of synaptic connectivity in the visual cortex. PMID- 8229807 TI - Task dependence of responses in first dorsal interosseous muscle to magnetic brain stimulation in man. AB - 1. The response of the first dorsal interosseous (1DI) muscle to non-invasive magnetic and scalp electrical stimulation of the brain have been investigated during performance of different manual tasks. 2. The six tasks tested required activation of the 1DI muscle, either in isolation (during abduction of the index finger) or as part of a more complex pattern of muscle synergies (e.g. during power grip). The level of 1DI EMG activity across tasks was kept constant by providing subjects with visual feedback of their muscle activity. 3. In every subject (n = 14) magnetic stimulation produced larger responses during performance of complex tasks than during the simple index abduction task. The pooled results from all subjects showed that four of the five complex tasks were associated with significantly larger 1DI responses (paired t test, P < 0.05). 4. These results were confirmed at the single motor unit level for nine motor units recorded from six subjects. Subjects were requested to produce a steady discharge of the same motor unit during performance of different tasks. The probability of motor unit discharge in response to magnetic stimulation was significantly greater during complex tasks (rotation or pincer grips) than during abduction. 5. Scalp electrical stimulation was performed in three subjects with the cathode at the vertex and the anode over the contralateral motor cortex. The pattern of response amplitudes in the different tasks tended to parallel that obtained for magnetic stimulation, but the task-related differences were smaller. 6. These results suggest that during performance of the different tasks, the corticospinal volleys evoked by magnetic stimulation may vary in amplitude. The task-related cortical mechanisms that may contribute to this variability are discussed. PMID- 8229808 TI - Nitric oxide-related vasodilator responses to parasympathetic stimulation of the submandibular gland in the cat. AB - 1. The extent to which parasympathetic vasodilator responses, in the submandibular gland of the cat, depend upon release of nitric oxide related (NO chi) or endothelium-derived relaxing factor (EDRF) within the gland has been investigated in anesthetized cats given N omega-nitro-L-arginine methyl ester (L NAME) which specifically blocks the synthesis of EDRF from arginine. 2. Close intra-arterial infusions of L-NAME (> or = 100 mg kg-1) produced a steady and significant rise in mean aortic pressure together with a steady increase in basal submandibular vascular resistance over the next 20-30 min, which persisted thereafter. 3. In cats pretreated with propranolol, to block beta-adrenoceptor mediated vasodilatation, salivation and vasodilatation in response to stimulation of the chorda-lingual nerve were reduced but not abolished by L-NAME (> or = 100 mg kg-1, I.A.). Subsequent administration of atropine (> or = 1 mg kg-1 I.V.) completely suppressed the secretory response and virtually eliminated the vascular response. 4. In cats pretreated with atropine (> or = 1.0 mg kg-1 I.V.) administration of L-NAME (> or = 100 mg kg-1 I.A.) effectively suppressed the vasodilator response to chorda-lingual stimulation at 2 Hz continuously, or at 20 Hz for 1 s at 10 s intervals. 5. Administration of L-NAME (> or = 100 mg kg-1 I.A.) effectively suppressed the submandibular vasodilator response to infusions of VIP (10 and 20 ng I.A.) and significantly reduced, but did not abolish that to acetylcholine (100 ng min-1 I.A.). 6. These results provide further support for the view that both acetylcholine and vasoactive intestinal polypeptide-like immunoreactivity (VIP) are released from the postganglionic parasympathetic nerve terminals and produce effects on the blood vessels in submandibular glands of the cat. They also provide evidence for a direct vascular action of acetylcholine, independent of NO chi, but VIP appears to act indirectly via NO chi formation. PMID- 8229809 TI - Identification and characterization of afferent periodontal A delta fibres in the cat. AB - 1. The presence and responsiveness of afferent periodontal A delta fibres was studied in pentobarbitone-anaesthetized adult cats. 2. Extracellular single fibre recordings were made from fine nerve filaments split from the proximally cut end of the inferior alveolar nerve. Periodontal nerve fibres were identified by constant current stimulus pulses applied via platinum wire electrodes inserted into the periodontal space of the lower canine tooth. 3. Of a total of 252 periodontal nerve fibres, 97 (37%) were classified as A delta fibres according to their conduction velocities (CV) (> 2.5 m s-1, < 30 m s-1) as determined by electrical stimulation of the periodontal ligament. The mean (+/- S.D.) conduction velocity was 11.0 +/- 7.7 m s-1 (n = 97; range: 2.6-28.2 m s-1). 4. A good exponential correlation (r = 0.85) was found between the electrical thresholds of the A delta fibres and their conduction velocities. 5. For four A delta fibres a complete stimulus-duration curve was determined. It followed rather well the I = I0/(1-et/tau) law, where I represents the stimulus amplitude, t the stimulus duration, I0 the rheobasic current and tau the time constant. 6. In the intact tooth none of the identified periodontal A delta fibres showed any ongoing activity in the absence of intentional stimulation. 7. The responses of sixteen electrically identified periodontal A delta fibres were tested by mechanical, thermal and chemical stimuli applied to the periodontal space. Seven of nine periodontal A delta fibres tested responded to mechanical forces applied to the tooth from different directions of which none could be activated by slight touch. A rudimentary directional sensitivity was seen. When a human tooth was stimulated by a mechanical stimulus of similar strength the sensation evoked was described as a dull, poorly localized pain. 8. Six periodontal A delta fibres were activated by heat and/or cold and/or chemical stimulation. Two of eight periodontal A delta fibres tested responded to heat and four of six A delta fibres tested responded to cold stimuli applied to the alveolar bone overlying the periodontal ligament; none of them responded to both types of thermal stimuli. Two of seven periodontal A delta fibres tested were activated by a saturated solution of potassium chloride applied locally to the periodontal ligament; two of these responded also to cold. 9. The response behaviour of periodontal afferent A delta fibres observed in this study suggests that they may play a role in periodontal nociception. PMID- 8229810 TI - A reassessment of mercury in silastic strain gauge plethysmography for microvascular permeability assessment in man. AB - 1. We have used non-invasive mercury in a silastic strain gauge system to assess the effect of pressure step size, on the time course of the rapid volume response (RVR) to occlusion pressure. We also obtained values for hydraulic conductance (Kf), isovolumetric venous pressure (Pvi) and venous pressure (Pv) in thirty-five studies on the legs of twenty-three supine control subjects. 2. The initial rapid volume response to small (9.53 +/- 0.45 mmHg, mean +/- S.E.M.) stepped increases in venous pressure, the rapid volume response, could be described by a single exponential of time constant 15.54 +/- 1.14 s. 3. Increasing the size of the pressure step, to 49.8 +/- 1.1 mmHg, gave a larger value for the RVR time constant (mean 77.3 +/- 11.6 s). 4. We propose that the pressure-dependent difference in the duration of the rapid volume response, in these two situations, might be due to a vascular smooth muscle-based mechanism, e.g. the veni arteriolar reflex. 5. The mean (+/- S.E.M.) values for Kf, Pvi and Pv were 4.27 +/- 0.18 (units, ml min-1 (100 g)-1 mmHg-1 x 10(-3), 21.50 +/- 0.81 (units, mmHg) and 9.11 +/- 0.94 (units, mmHg), respectively. 6. During simultaneous assessment of these parameters in arms and legs, it was found that they did not differ significantly from one another. 7. We propose that the mercury strain gauge system offers a useful, non-invasive means of studying the mechanisms governing fluid filtration in human limbs. PMID- 8229811 TI - Multiple mechanisms of picrotoxin block of GABA-induced currents in rat hippocampal neurons. AB - 1. We have examined the effect of picrotoxin on GABA-induced currents in dissociated rat hippocampal neurons. In addition, we used the putative picrotoxin receptor antagonist, alpha-isopropyl-alpha-methyl-gamma-butyrolactone (alpha IMGBL), and the picrotoxin agonist, beta-ethyl-beta-methyl-gamma-butyrolactone (beta EMGBL) to explore the mechanisms of picrotoxin's interaction with the GABA Cl- receptor-ionophore complex. 2. The picrotoxin block of GABA current was use dependent, suggesting that the site of picrotoxin block is exposed by the conformational change initiated by GABA binding to the receptor. 3. The alkyl substituted butyrolactone antagonist, alpha IMGBL, selectively blocked the use dependent mechanism of picrotoxin effect. After the apparent complete inhibition of the use-dependent effect, there was a residual picrotoxin effect that was independent of the time or concentration of GABA application. This indicates that the picrotoxin block of the GABA current is mediated by two different mechanisms. alpha IMGBL influences just one of these mechanisms. 4. The picrotoxin receptor agonist, beta EMGBL, exclusively blocked the GABA current in a use-dependent manner. Consistent with a use-dependent mechanism, the rate of onset of block increased with GABA concentration. Surprisingly, the fraction of GABA current block decreased with increasing GABA concentration. 5. These results suggest that the relationship of picrotoxin and gamma-butyrolactones with the GABA-Cl- receptor-ionophore is quite complex. They are consistent with at least two possible models of agonist-antagonist interactions. Both cases require different antagonist affinities for the various kinetic states of the GABA-Cl- receptor ionophore. However, there is no need to require that either picrotoxin or beta EMGBL acts as an open channel blocker. PMID- 8229812 TI - Intracellular ATP modifies the voltage dependence of the fast transient outward K+ current in Lymnaea stagnalis neurones. AB - 1. The action of intracellular ATP on the fast transient outward K+ current (A current) was studied in dialysed voltage-clamped Lymnaea stagnalis neurones. 2. When introduced intracellularly in millimolar concentrations ATP caused a shift of the steady-state inactivation curve along the voltage axis in the direction of positive potentials and decreased A-current at all test voltages. 3. Intracellular treatment with an inhibitor of ATP synthesis, sodium arsenate, led to the opposite changes. The action of arsenate was not reversed upon its removal. After wash-out of arsenate ATP restored the initial voltage dependence. 4. Addition of Mg2+ to the solution weakened the action of ATP in proportion to the Mg2+: ATP concentration ratio. On the other hand, in neurones pretreated with arsenate, Mg2+ did not affect the ATP action. 5. When a mixture of glycolytic substrates was applied after arsenate wash-out the activation and inactivation curves shifted towards positive voltages. A substrate of oxidative phosphorylation was ineffective in the same conditions. 6. Non-hydrolysable analogues of ATP, adenosine-5'-O-gamma-thiotriphosphate and adenylyl imidodiphosphate, did not mimic the ATP action. This means that the ATP effect is mediated by some enzymatic process(es). 7. Elevation of total cytosolic Ca2+ concentration as well as intracellular application of agents increasing intracellular free Ca2+ reduced A-current amplitude but failed to alter its voltage dependence. Therefore, ATP action cannot be related to activation of Ca2+ transport. 8. Treatment of the neurones with alkaline phosphatase evoked a shift of the inactivation voltage dependence towards hyperpolarizing potentials and increased the A-current amplitudes at all test voltages. 9. The data indicate that a change in intracellular ATP concentration modulates the A-current voltage dependence. The effect of ATP is probably the result of phosphorylation of a channel protein or some associated proteins, but lowering of free Mg2+ concentration cannot be excluded. The possible physiological significance of the phenomenon is discussed. PMID- 8229813 TI - Mechanical factors in the initiation of eccentric contraction-induced injury in rat soleus muscle. AB - 1. Mechanical factor(s) associated with the initiation of eccentric contraction induced muscle injury were investigated in isolated rat soleus muscles (n = 180; 42 protocols with 4-6 muscles per protocol). Five eccentric contractions were performed with 4 min between contractions. Three levels of peak eccentric contraction force (100, 125 and 150% of pre-injury maximal isometric tetanic tension, P0), length change (0.1, 0.2 and 0.3 muscle length, L0) and lengthening velocity (0.5, 1.0 and 1.5 L0/s) were utilized. Force was varied with stimulation frequency (10-150 Hz). The eccentric contractions were initiated at muscle lengths of 0.85 or 0.90 L0. Following the fifth eccentric contraction, the muscle was incubated in Krebs-Ringer buffer for 60 min. Peak isometric twitch tension (PT), P0, maximal rate of tension development (+ dP/dt), maximal rate of relaxation (-dP/dt), and creatine kinase (CK) release were measured prior to the five eccentric contractions and at 15 min intervals during the incubation period. Total muscle [Ca2+] was measured after 60 min incubation. 2. The mean (+/- S.E.M.) initial decline in P0 for the muscles performing the most injurious protocol was 13.6 +/- 4.8% (n = 6); P0 in control muscles immediately following performance of five isometric contractions was elevated 1.2 +/- 1.0% (n = 8). These means were different at probability, p = 0.005. Mean [ATP] in muscles immediately following the isometric control and most injurious protocols, respectively, were 16.30 +/- 1.49 and 19.84 +/- 1.38 mumol/g dry wt (p = 0.229). 3. Decrements in P0, PT, +dP/dt, and -dP/dt immediately after the injury protocol were related most closely to the peak forces produced during the eccentric contractions; greater initial declines in P0, +dP/dt and -dP/dt were also observed at higher lengthening velocities independent of peak force. Slow declines in P0 and -dP/dt during the 60 min incubation following the injury protocol were greatest for muscles performing contractions at the longer initial length. CK release was independent of all mechanical factors with the exception of lengthening velocity. CK activity at 45 and 60 min into the incubation period was greater for muscles lengthened at the highest velocity used (1.5 L0/s). Mean total muscle [Ca2+] for muscles performing the eccentric contractions was elevated by 38% over isometric control muscles but the elevation was unrelated to any of the four mechanical factors. 4. These data support the hypothesis that eccentric contraction-induced injury is initiated by mechanical factors, with muscle tension playing the dominant role.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229814 TI - Materials fatigue initiates eccentric contraction-induced injury in rat soleus muscle. AB - 1. The initiation of exercise-induced muscle injury is thought to be the result of high tensile stresses produced in the muscle during eccentric contractions. Materials science theory suggests that high tensile stresses could initiate the injury during the first eccentric contraction (normal stress theory) or after multiple eccentric contractions (materials fatigue). It was the objective of this study to investigate the two possibilities. 2. Rat soleus muscles (n = 66; 11 protocols with 6 muscles per protocol) were isolated, placed in an oxygenated Krebs-Ringer buffer at 37 degrees C, and baseline measurements were made. The muscle then performed an injury protocol which consisted of between zero and ten eccentric contractions (muscle starting length = 0.90 soleus muscle length, L0; length change = 0.25 L0; velocity = 1.5 L0/s; peak force = 180% maximal isometric tetanic tension (P0); time between contractions = 4 min; total duration of the injury protocol = 40 min). At the end of the injury protocol, the muscle was incubated in buffer for 1 h; every 15 min, an isometric twitch and tetanus were performed and lactate dehydrogenase (LDH) release was measured. Total muscle [Ca2+] was measured at the end of the incubation. 3. Change-point regression analysis indicates that at 0 min into the incubation, declines in P0, maximal rate of tension development (+dP/dt), maximal rate of relaxation (-dP/dt), and muscle stiffness (dP/dx) became significantly greater after eight eccentric contractions (p < or = 0.05). No relation was found between the number of eccentric contractions performed and the LDH activity at 0 min into the incubation, although after 60 min of incubation, LDH activity in the buffer was linearly related to eccentric contraction number (p = 0.01). There was no relationship between total muscle [Ca2+] and eccentric contraction number. These findings support the materials fatigue hypothesis of exercise-induced muscle injury. PMID- 8229815 TI - Ca(2+)-dependent inactivation of Ca2+ current in Aplysia neurons: kinetic studies using photolabile Ca2+ chelators. AB - 1. The kinetics and sensitivity of the Ca(2+)-dependent inactivation of calcium current (ICa) were examined in intact cell bodies from the abdominal ganglion of Aplysia californica under two-electrode voltage clamp. 2. Rapid changes in the level of intracellular free calcium ([Ca2+]i) were generated at the cell surface by photolytic release of Ca2+ (nitr-5 and dimethoxy nitrophen) or Ca2+ buffer (diazo-4). 3. Diazo-4 increased ICa by 10-15% and slowed the rate of ICa decay when photolysed before a test pulse or between a prepulse and a test pulse. The predominant effect of further light flashes was to increase the amount of non inactivating current (I infinity) remaining at the end of long (> 1 s) depolarizing pulses. 4. A rapid increase in [Ca2+]i buffering during ICa inactivation did not cause a rapid recovery of current but merely reduced the rate and extent of subsequent inactivation. This effect was not seen when Ba2+ was the charge carrier. 5. Photolytic release of Ca2+ from nitr-5 produced estimated Ca2+ jumps of 3-4 microM at the front surface of the cell but failed to augment inactivation either before or during ICa. In contrast, photolysis of DM nitrophen 10-90 ms before the test pulse decreased peak ICa by about 30%. A flash given during ICa rapidly blocked 41 +/- 3% of peak current with a time constant of 3-4 ms at 17 degrees C. Similar results were seen with the barium current (IBa). 6. Microinjection of the potent phosphatase inhibitor microcystin-LR (5 microM) had variable effects on ICa inactivation and augmented the cyclic AMP induced depression of the delayed rectifier (IK(V) by forskolin (100 microM) and 3-isobutyl-1-methylxanthine (IBMX; 200 microM). 7. Full recovery from inactivation measured in two-pulse experiments took at least 20 s. This slow recovery process was unaffected by increases in intracellular cyclic AMP elicited by direct injection or by bath application of forskolin and IBMX. It was also unaffected by decreases in cyclic AMP induced by injecting 2',5'-dideoxyadenosine (1 mM) or bath application of the Rp isomer of cyclic adenosine 3',5' monophosphothioate (Rp-cAMPS; 200 microM). 8. A 'shell' model relating submembrane Ca2+ to inactivation was inconsistent with the experimental results since it greatly overestimated the effects of diazo-4 and predicted significant inactivation by nitr-5 photolysis. 9. A model linearly relating [Ca2+]i in a single Ca2+ channel 'domain' to inactivation more closely matched the experimental results with diazo-4 and DM-(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229816 TI - Effects of deoxygenation on active and passive Ca2+ transport and cytoplasmic Ca2+ buffering in normal human red cells. AB - 1. The effects of deoxygenation on cytoplasmic Ca2+ buffering, saturated Ca2+ extrusion rate through the Ca2+ pump (Vmax), passive Ca2+ influx and physiological [Ca2+]i level were investigated in human red cells to assess whether or not their Ca2+ metabolism might be altered by deoxygenation in capillaries and venous circulation. 2. The study was performed in fresh human red cells maintained in a tonometer either fully oxygenated or deoxygenated. Cytoplasmic Ca2+ buffering was estimated from the equilibrium distribution of 45Ca2+ induced by the divalent cation ionophore A23187 and the Vmax of the Ca2+ pump was measured either by the Co(2+)-exposure method or following ionophore wash-out. The passive Ca2+ influx and physiological [Ca2+]i were determined in cells preloaded with the Ca2+ chelator benz-2 and resuspended in autologous plasma. 3. Deoxygenation increased the fraction of ionized Ca2+ in cell water by 34-74% and reduced the Vmax of the Ca2+ pump by 18-32%. 4. To elucidate whether or not these effects were secondary to deoxygenation-induced pH shifts, the effects of deoxygenation on cell and medium pH, and of pH on cytoplasmic Ca2+ binding and Ca2+ pump Vmax in oxygenated cells were examined in detail. 5. Deoxygenation generated large alkaline pH shifts that could be explained if the apparent isoelectric point (pI) of haemoglobin increased by 0.2-0.4 pH units in intact cells, consistently higher than the value of 0.15 reported for pure haemoglobin solutions. 6. In oxygenated cells, the fraction of ionized cell calcium, alpha, was little affected by pH within the 7.0-7.7 range. Ca2+ pump Vmax was maximal at a medium pH of about 7.55. Comparison between pH effects elicited by HCl-NaOH additions and by replacing Cl- with gluconate suggested that Vmax was inhibited by both internal acidification and external alkalinization. Since deoxygenation alkalinized cells and medium within a range stimulatory for Vmax, the inhibition observed was not due to pH. 7. There was no significant effect of deoxygenation on passive Ca2+ uptake, or steady-state physiological [Ca2+]i level. 8. The deoxygenation-induced reduction in Ca2+ binding capacity may result from the increased protonation of haemoglobin on deoxygenation and from binding of 2,3-diphosphoglyceric acid (2,3-DPG) and ATP to deoxyhaemoglobin; inhibition of the Ca2+ pump may result from shifts in the [Mg2+]i/[ATP]i ratio away from a near optimal stimulatory value in the oxygenated state. PMID- 8229817 TI - Observations on human tactile directional sensibility. AB - 1. The ability to tell the direction of a motion across the skin deserve attention for being an easily observed function which provides a sensitive test for disturbances of the peripheral and central nervous systems. The mode of operation, on the other hand, of this tactile directional sensibility is still uncertain. 2. The dependence of directional sensibility on the contact load and distance of movement of a blunt metal tip, has now been determined for the skin of the forearm of normal subjects with the two-alternative forced-choice method. The testing was done under two conditions: elbow bent or straight. Straightening of the arm always reduced the accuracy of the directional sensibility. It also caused measurable changes of cutaneous mechanical properties, which presumably decreased the reliability of afferent information about lateral distension. 3. The average accuracy of the directional sensibility was found to be correlated linearly to the logarithm of the contact load, and straightening of the arm decreased the accuracy for each load by corresponding amounts. Similar relationships were found between the accuracy and the distance of movement. 4. Straightening of the arm did not cause any significant average reduction of the contact threshold for point stimulation of the same receptive field. A consistently lowered contact sensitivity, however, was observed for some of the subjects, which may have contributed to the reduction of the directional sensibility in these cases. 5. Correct directional estimations of the movement of the metal tip were obtained for a distance which was a fifth of the shortest distance for a corresponding estimation of the movement of a frictionless stimulus. The findings thus indicated that the friction between a moving object and the underlying skin, which can be mediated via stretch-sensitive cutaneous receptors, is critical for the determination of its direction of motion. 6. The present observations and previous observations by various authors are suggested to indicate that typical tactile directional sensibility depends on parallel processing of direction-selective data, and spatial data expressed as a function of time. PMID- 8229818 TI - Changes in myoplasmic pH and calcium concentration during exposure to lactate in isolated rat ventricular myocytes. AB - 1. We investigated the mechanisms involved in the rise of myoplasmic calcium concentration ([Ca2+]i) when isolated rat ventricular myocytes were exposed to lactate. The intracellular pH (pHi) and [Ca2+]i were measured using the fluorescent indicators 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) and fura-2, respectively. Cell shortening was used as a measure of contractile performance. 2. Exposure to 20 mM lactate at the normal extracellular pH (pHo 7.4) for 10 min caused the pHi to fall rapidly by 0.24 pH units and cell shortening was reduced. Thereafter, pHi partially recovered by 0.16 pH units, which was paralleled by a recovery of shortening. 3. Exposure to lactate at a reduced extracellular pH (pHo 6.4) induced a very large acidosis of 0.70 pH units and cell shortening was abolished. During maintained exposure to lactate the pHi remained constant and cell shortening did not recover. 4. Application of Na(+)-H+ exchanger inhibitors, amiloride or ethylisopropyl-amiloride (EIPA), abolished the recovery of pHi and shortening during maintained exposure to lactate at pHo 7.4 and caused an additional acidosis during maintained application of lactate at pHo 6.4. 5. Application of lactate at both the normal and reduced pHo resulted in a rapid, followed by a slower, rise in [Ca2+]i. The diastolic and systolic [Ca2+]i and the amplitude of the systolic rise in the [Ca2+]i (the Ca2+ transient) all increased in both the rapid and the slow phase. 6. When lactate was applied at pHo 7.4, in the presence of EIPA, the initial rise of [Ca2+]i still occurred but the slower increase was abolished. This suggests an involvement of the Na(+)-H+ exchanger in the slower rise of [Ca2+]i. 7. In conclusion, the Na(+)-H+ exchanger is an important regulator of pHi during a lactate-induced intracellular acidosis. The rise of [Ca2+]i involves at least two mechanisms: (i) a rapid component which may represent reduced myoplasmic Ca2+ buffering, impaired Ca2+ removal by the sarcoplasmic reticulum or a direct inhibitory effect of protons on the Na(+)-Ca2+ exchanger; (ii) a slower component linked to stimulation of Na(+)-H+ exchanger which causes an increased [Na+]i and stimulates the Na(+)-Ca2+ exchanger, resulting in an enhanced Ca2+ influx. PMID- 8229819 TI - The regulation of presynaptic inhibition during co-contraction of antagonistic muscles in man. AB - 1. The size of the soleus, tibialis anterior and medial gastrocnemius H reflexes were measured at different levels of EMG activity in ankle plantar- and dorsiflexors. At similar levels of EMG activity in the agonist muscle, the soleus and tibials anterior H reflexes were found to be smaller during co-contraction than during an isolated agonist contraction. In contrast to this, the medial gastrocnemius H reflex had the same size during co-contraction as during an isolated plantarflexion at comparable EMG levels. 2. Peaks occurred at monosynaptic reflex latency in the post-stimulus time histogram (PSTH) of firing of soleus and tibialis anterior motor units following stimulation of their respective motor nerves. These homonymous Ia peaks were found to be smaller during co-contraction than during isolated agonist contraction, whereas that of medial gastrocnemius motor units was unchanged. As the monosynaptic peak in the PSTH may give an indirect measure of the size of the monosynaptic EPSP in the motoneurone, it is suggested that these findings reflect a differential control of presynaptic inhibition of soleus and tibialis anterior motor units with respect to medial gastrocnemius motor units. 3. The monosynaptic Ia facilitation of the soleus H reflex evoked by a stimulation of the femoral nerve, was also found to be smaller during co-contraction of tibials anterior and soleus than during isolated plantarflexion. This suggests that presynaptic inhibition of heteronymous Ia fibres from the quadriceps muscle projecting to soleus motoneurones is regulated in the same way as presynaptic inhibition of homonymous Ia fibres during co-contraction. 4. The femoral nerve-induced monosynaptic facilitation of the soleus H reflex was found to decrease with increasing tonic levels of both co-contraction and isolated dorsi- and plantarflexion. The amount of facilitation decreased to the same degree with increasing levels of dorsiflexion and co-contraction. For the same level of EMG in the dorsiflexors there was thus no difference in the amount of facilitation whether the dorsiflexors were activated separately or in a co-contraction. 5. Whereas the soleus H reflex increased in size 50 ms prior to the onset of a dynamic plantarflexion, it did not change prior to the onset of a co-contraction. The femoral nerve-induced facilitation increased prior to the plantarflexion, but decreased prior to the co-contraction.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229820 TI - Saccade-vestibulo-ocular reflex co-operation and eye-head uncoupling during orientation to flashed target. AB - 1. Eye-head co-ordination in the horizontal plane was studied in four human subjects using two successive flashes in the same direction, either increasing in eccentricity (IE), or decreasing in eccentricity (DE). 2. Results showed that for both conditions, head movements preceded eye movements and were typically longer or followed by a slow gaze movement. This slow movement was due to a vestibulo ocular reflex gain of less than one. Gaze accuracy was achieved by small head movement adjustments. 3. Gaze movement to an IE stimulus had a staircase pattern, and to a DE stimulus, a pulse-step pattern or one gaze saccade to the final flash eccentricity. 4. In some cases, however, in response to a DE stimulus, the eye and head movements were directed to different displacements (dissociation); i.e. the head movement started towards the first flash eccentricity with a concomitant eye saccade to the second flash eccentricity. When this occurred, gaze movement did not resemble a pulse-step pattern. 5. It is suggested that non-visually orienting gaze is driven mainly by head movement. Eye and head movements can be either tightly coupled or dissociated, depending on the stimulus pattern. PMID- 8229821 TI - Differences in adaptation between on- and off-centre ganglion cells and rod mediated cone sensitization in cat retina. AB - 1. Response properties of on- and off-centre retinal ganglion cells were investigated in cats. The stimulus parameters were selected so as to demonstrate interactions between the rod and the cone systems. 2. Response versus log stimulus intensity (R-log I) functions were determined for the receptive field centres while both test stimulus irradiance and the background illumination were varied over a range up to 7 log units. In order to determine the course of adaptation to chromatic stimuli, threshold versus intensity (t.v.i.) functions were measured over a wide range of adaptation levels. 3. An increase in background illuminance produced a shift of the R-log I functions to higher irradiances of test stimuli in most ganglion cells, indicating a desensitization of the centre response in the presence of background lights. Using test stimuli which most efficiently stimulate the rods (501 nm), clear differences could be seen in the adaptation behaviour of on- and off-centre ganglion cells. Chromatic backgrounds (blue-green and orange) reduced the responses of off-centre cells more than those of on-centre cells (the difference between them amounting to as much as 2 log units). Simultaneously, equivalent t.v.i. functions had significantly steeper slopes (0.94 and 1.1) in the linear proportions of off centre cells compared to on-centre cells (0.76 and 0.75) under light levels mediated by rods. Such differences were not observed when a test stimulus of 575 nm was used which resulted primarily in stimulating the long-wavelength cone (L cone) system. 4. In a subpopulation of off-centre cells (20% of the total number of off-centre cells recorded), a strikingly different adaptation behaviour was observed. Here, the presentation of a dim short-wavelength background produced a shift of R-log I functions to lower test stimulus irradiances. The receptive field centre became even more sensitive, by up to 1.5 log units, in the presence of dim adapting backgrounds rather than in the dark-adapted state. Accordingly, the t.v.i. function did not increase monotonically but showed a 'dip' in the presence of dim backgrounds. Only at photoic levels, the t.v.i. functions revealed a response behaviour similar to the other ganglion cells. The sensitization with dim backgrounds was only observed in the case of test stimuli designed to stimulate the cone system (575 nm) and in the presence of a rod adapting blue-green background.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229822 TI - Effects of osmolality and ionic strength on the mechanism of Ca2+ release in skinned skeletal muscle fibres of the toad. AB - 1. The effects of increased osmolality and ionic strength on the mechanism of Ca2+ release were examined in mechanically skinned skeletal muscle fibres of the toad at 23 degrees C. Ca2+ release was induced by depolarizing the transverse tubular (T-) system by ionic substitution. 2. Increasing the osmolality of the 'myoplasmic' solution about four times (to 955 mosmol/kg), by addition of 700 mM sucrose to the standard potassium (K-)HDTA solution (HDTA: hexamethylenediamine tetraacetate), only depressed the depolarization-induced response by about 46%. Much of this decrease could be attributed to a reduction in the Ca(2+) sensitivity of the contractile proteins at this high osmolality. 3. Addition of > 400 mM sucrose itself often induced substantial Ca2+ release and a transient tension response. This 'spontaneous' release was (a) greatly enhanced when the sarcoplasmic reticulum (SR) had been heavily loaded with Ca2+, (b) little affected by inactivation of the voltage sensors by prolonged or permanent depolarization of the T-system and (c) blocked by Ruthenium Red (10 microM). 4. When both the osmolality and ionic strength were increased, by increasing the K HDTA concentration, the depolarization-induced force was greatly reduced (to 35% at 818 mosmol/kg and 5% at 1095 mosmol/kg). Most of this reduction could be directly attributed to the substantially reduced maximum force and Ca2+ sensitivity of the contractile apparatus. 5. The small amount of releasable Ca2+ remaining in the SR after a single depolarization in a high-HDTA solution with 1 mM EGTA (to chelate the released Ca2+), indicated that depolarization could still elicit massive Ca2+ release at high ionic strength and osmolality (at 1 mM free Mg2+). 6. In contrast, when the total Mg2+ and ATP concentrations were raised about threefold (free [Mg2+] increased 2.7-fold) along with the osmolality and ionic strength, the ability of depolarization to elicit Ca2+ release was greatly hindered. 7. Osmotic compression of the skinned fibres to their in situ diameter by addition of 4% polyvinylpyrrolidone (PVP-40), substantially potentiated the depolarization-induced force responses, due partly to an increase in the Ca(2+) sensitivity of the contractile apparatus. 8. These results indicate how increased intracellular osmolality, ionic strength and [Mg2+] produce the transient contraction and subsequent inhibition of tetanic tension in intact muscle fibres exposed to hypertonic solutions. PMID- 8229823 TI - On the role of G protein activation and phosphorylation in desensitization to acetylcholine in guinea-pig atrial cells. AB - 1. The ACh-activated K+ current (IK,ACh) has been investigated in guinea-pig atrial cells at 36 degrees C using the whole-cell patch-clamp technique. 2. During an exposure to ACh, IK,ACh faded as a result of desensitization. Throughout the fade of the current, the current reversed at EK and showed inward going rectification. The fade was, therefore, the result of a genuine decrease in IK,ACh. 3. The onset of desensitization (as judged by the fade of IK,ACh) was biphasic and the time constants of the fast and slow phases of desensitization were 1.58 +/- 0.14 (n = 16) and 148.2 +/- 12.8 s (n = 18) respectively. Recovery from the fast and slow phases of desensitization (after 30 s and 5 min exposures to ACh respectively) occurred with time constants of 52 and 222 s respectively. This suggests that two processes are involved in desensitization. 4. The Q10 of the rate constant of the fast phase of desensitization was 2.2 +/- 0.3 (n = 6). 5. Intracellular perfusion with guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) or extracellular perfusion with AlF4- were used to bypass the muscarinic receptor and trigger IK,ACh by directly activating the G protein, GK, that links the muscarinic receptor to the K+ channel. Both GTP gamma S and AlF4- activated a current with the same reversal potential and the same degree of inward-going rectification as the ACh-activated current. 6. Desensitization still occurred when the muscarinic receptor was bypassed and IK,ACh was triggered by direct activation of GK with either GTP gamma S or AlF4-. This suggests that desensitization is, in part, the result of a modification of either GK or the K+ channel. 7. Activation of the muscarinic receptor by ACh resulted in greater desensitization than direct activation of GK; at the end of a 5 min exposure to ACh, current was only 22 +/- 1% (n = 19) of its peak value, whereas, after direct activation of GK by GTP gamma S for 5 min, current was 42 +/- 6% (n = 5) of its peak value. This suggests that desensitization also involves the muscarinic receptor. 8. When cells were perfused with GTP gamma S, the fast phase of desensitization could still occur, but the slow phase was reduced. This suggests that the fast phase involves GK or the K+ channel, whereas the slow phase involves the muscarinic receptor.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229824 TI - Identification of vagal sensory receptors in the rat lung: are there subtypes of slowly adapting receptors? AB - 1. We studied the characteristics of pulmonary sensory receptors whose afferent fibres are in the left vagus nerve of opened-chest rats. The activity of these receptors was recorded during mechanical ventilation approximating eupnoea, as well as during deflation, stepwise inflations and constant-pressure inflations of the lungs. Data were also collected from closed-chest rats and analysed separately. 2. Ninety-four per cent of receptors were located in the ipsilateral lung or airways with the remainder in the contralateral lung. 3. Not only were slowly adapting receptors (SARs) the most abundant pulmonary receptors but 21% of them were either exclusively or predominantly active during the deflationary phase of the ventilatory cycle. Deflationary units were found in opened- and closed-chest rats. The average conduction velocity for all fibres innervating SARs averaged 29.7 m s-1. 4. We found rapidly adapting receptors (RARs) to be extremely rare in the rat. Their activity was sparse and irregular. The conduction velocities of fibres innervating RARs averaged 12.3 m s-1. 5. Far more abundant than RARs in the remaining population of pulmonary fibres were C fibres. They were observed to have an average conduction velocity of 2.1 m s-1, base level activity which was irregular and a high pressure threshold of activation and were stimulated by intravenous capsaicin injection. 6. Notable differences exist between pulmonary receptors in rats and those reported in other species. The variations include the abundant existence of intrapulmonary SARs with exclusively deflationary modulation and the rarity of RARs. We also encountered C fibres which have not previously been described systematically in the rat. PMID- 8229825 TI - Effects of repeated tetanic stimulation on excitation-contraction coupling in cut muscle fibres of the frog. AB - 1. The effects of prolonged intermittent fatiguing stimulation were studied on various steps of excitation-contraction (E-C) coupling in cut single frog muscle fibres using the triple Vaseline voltage clamp and the fluorescent Ca2+ indicator rhod-2. 2. There were two phases of changes in amplitude of Ca2+ transients during fatiguing stimulation: first a 5-10% increase, then a larger decrease. The decrease in amplitude of Ca2+ transients was accompanied by a slowing down of the rate of decay of the transients and by an increase in resting [Ca2+]. 3. A complete recovery of both amplitude and time course of Ca2+ transients as well as of the resting [Ca2+] occurred within 1-3 min after cessation of fatiguing stimulation. 4. The changes in Ca2+ release signals during fatiguing stimulation were accompanied by decreases in the amplitude and the rate of decay of the action potentials as well as by a decrease in resting potential. However, these alterations are not likely to contribute to fatigue significantly, since fibres stimulated under voltage-clamp conditions, when the T-tubule voltage sensor is activated directly by applied voltage steps, showed similar fatiguability to fibres stimulated by action potentials under current-clamp conditions. 5. Simultaneous measurements of intramembrane charge movement and [Ca2+] revealed that the decrease in sarcoplasmic reticulum (SR) Ca2+ release during fatiguing stimulation is not accompanied by any significant change in charge movement. 6. These results suggest that fatigue caused by repeated tetanic stimulation develops primarily at the level of SR Ca2+ release with only small possible additional effects at the level of membrane excitability and action potential propagation along the surface/T-tubule membrane. The T-tubule voltage sensor with this type of stimulation is virtually fatigue resistant. PMID- 8229826 TI - Mechanoreceptors around the tooth evoke inhibitory and excitatory reflexes in the human masseter muscle. AB - 1. The reflex responses evoked in the human masseter muscle by controlled mechanical stimulation of an incisor tooth were examined electromyographically. The stimuli were (slow) pushes and (brisk) taps of about 0.5-3 N peak force, applied orthogonally to the labial surface. 2. The brisk taps elicited a short latency inhibitory reflex that was often followed by an excitatory peak, as has been described earlier. The inhibition increased as the taps became stronger. 3. Slow pushes evoked a long-latency, primarily excitatory response. The excitation increased with stronger, faster rise-time pushes; however, with the stronger stimuli, the short-latency inhibitory response often became evident before the onset of the excitation. 4. The reflex responses to 3 N pushes and 2 N taps were abolished when the receptors around the tooth were blocked with local anaesthetic, indicating that the response was elicited from receptors located within the periodontal area. 5. Prolonged, rapid-onset displacements evoked a complex reflex response that combined the characteristics of the taps and the pushes. 6. The most likely explanation for the different responses evoked by the pushes and taps is that the patterns of afferent activity elicited by the slow and fast tooth displacements activated different interneuronal pathways to motoneurones. 7. The inhibitory response to taps is essentially a protective reflex which probably serves to reduce the activity of the jaw-closing muscles when one bites unexpectedly on hard objects. It is suggested that the excitatory response may contribute to the muscle activity required to hold food between the teeth during chewing, or may act as a load compensation reflex to control chewing force. PMID- 8229827 TI - Pulmonary stretch receptor afferents activate excitatory amino acid receptors in the nucleus tractus solitarii in rats. AB - 1. The goal of the present study was to identify potential neurotransmitter candidates in the Breuer-Hering (BH) reflex pathway, specifically at synapses between the primary afferents and probable second-order neurones (pump cells) within the nucleus tractus solitarii (NTS). We hypothesized that if activation of specific receptors in the NTS is required for production of the BH reflex, then (1) injection of the receptor agonist(s) would mimic the reflex response (apnoea), (2) injection of appropriate antagonists would impair the apnoea produced by either lung inflation or agonist injection, and (3) second-order neurones in the pathway would be excited by either lung inflation or agonists while antagonists would prevent the response to either. 2. Studies were carried out either in spontaneously breathing or in paralysed, thoracotomized and ventilated rats in which either diaphragm EMG or phrenic nerve activity, expired CO2 concentration and arterial pressure were continuously monitored. The BH reflex was physiologically activated by inflating the lungs. 3. Pressure injections (0.03-15 pmol) of selective excitatory amino acid (EAA) receptor agonists, quisqualic acid (Quis) and N-methyl-D-aspartic acid (NMDA) into an area of the NTS shown previously to contain neurones required for production of the BH reflex produced dose-dependent apnoeas that mimicked the response to lung inflation. Injection of substance P (0.03-4 pmol) did not alter baseline respiratory pattern. 4. Injections of the EAA antagonists, kynurenic acid (Kyn; 0.6-240 pmol), 6-cyano-7-nitro-quinoxaline-2,3-dione (CNQX) or 6,7 dinitroquinoxaline-2,3-dione (DNQX) into the BH region of the NTS reversibly impaired the apnoea produced by lung inflation. All three antagonists reduced or abolished the apnoeas resulting from injection of Quis or NMDA, and slowed baseline respiratory frequency. In contrast, injections of the highly selective NMDA receptor antagonist, D-2-amino-5-phosphonovaleric acids (AP5), in doses sufficient to block the apnoeic response to NMDA, neither altered the reflex apnoea evoked by lung inflation nor the baseline respiratory pattern. 5. Pump cells located within the BH region were excited by pressure injections of the broad spectrum EAA agonist, DL-homocysteic acid (DLH). Kyn reversibly blocked the excitation of pump cells in response to either lung inflation or DLH injection. 6. These findings suggest that EAAs mediate primary afferent excitation of second order neurones in the Breuer-Hering reflex pathway, primarily through the activation of non-NMDA EAA receptor subtypes. PMID- 8229828 TI - Visual transduction in human rod photoreceptors. AB - 1. Photocurrents were recorded with suction electrodes from rod photoreceptors of seven humans. 2. Brief flashes of light evoked transient outward currents of up to 20 pA. With increasing light intensity the peak response amplitude increased along an exponential saturation function. A half-saturating peak response was evoked by approximately sixty-five photoisomerizations. 3. Responses to brief dim flashes rose to a peak in about 200 ms. The waveform was roughly like the impulse response of a series of four to five low-pass filters. 4. The rising phases of the responses to flashes of increasing strength were found to fit with a biochemical model of phototransduction with an 'effective delay time' and 'characteristic time' of about 2 and 800 ms, respectively. 5. Spectral sensitivities were obtained over a wavelength range from 380 to 760 nm. The action spectrum, which peaked at 495 nm, followed the template described for photoreceptors in the macaque retina. Variation between rods in the position of the spectrum on the wavelength axis was small. 6. The scotopic luminosity function derived from human psychophysical experiments was found to agree well with the measured rod action spectrum after adjustments were made for lens absorption and photopigment self-screening in the intact eye. 7. Responses to steps of light rose monotonically to a maintained level, showing little or no relaxation. Nevertheless, the relationship between light intensity and steady state response amplitude was shallower than that expected from simple response saturation. This is consistent with an adaptation mechanism acting on a rapid time scale. 8. Flash sensitivity fell with increasing intensities of background light according to Weber's law. Sensitivity was reduced twofold by lights evoking about 120 photoisomerizations per second. Background lights decreased the time to peak and the integration time of the flash response by up to 20%. PMID- 8229829 TI - Adaptation of cat motoneurons to sustained and intermittent extracellular activation. AB - 1. The main purpose of this study was to quantify the adaptation of spinal motoneurons to sustained and intermittent activation, using an extracellular route of stimulating current application to single test cells, in contrast to an intracellular route, as has been used previously. In addition, associations were tested between firing rate properties of the tested cells and other type (size) related properties of these cells and their motor units. 2. Motoneurons supplying the medial gastrocnemius muscle of the deeply anaesthetized cat were stimulated for 240 s with microelectrodes which passed sustained extracellular current at 1.25 times the threshold for repetitive firing. Many cells were also tested following a rest period with intermittent 1 s current pulses (duration 600 ms) at the same relative stimulus strength. Cell discharge was assessed from the EMG of the motor unit innervated by the test neuron. The motoneurons and their motor units were assigned to four categories (i.e. types FF, FR, S and F; where F = FF + FR) based on conventional criteria. In all, twenty F (16 FF, 4 FR) and fourteen S cells were studied with sustained stimulation. Thirty of these cells (17 F, 13 S) and an additional two cells (1 F, 1 S) were studied with intermittent stimulation. 3. The mean threshold current required for sustained firing for a period of > or = 2 s was not significantly different for F and S cells. However, most of the other measured parameters of motoneuron firing differed significantly for these two cell groups. For example, at 1.25 times the threshold current for repetitive firing, the mean firing duration in response to 240 s of sustained activation was 123 +/- 88 s (+/- S.D.) for F cells vs. 233 +/- 19 s for S cells. These values were significantly longer than those from a comparable, previously reported study that employed intracellular stimulation. With intermittent stimulation, the firing durations of F and S cells were not significantly different from each other. 4. All cells exhibited a delay from the onset of current to the first spike, followed by a brief accelerating discharge that was followed by a slower drop in firing rate. Some cells (21 of 34 with sustained activation; 20 of 32 with intermittent) exhibited doublet discharges (interspike intervals < or = 10 ms) that were intermingled with the more predominant singlet discharges. Doublets were more common in the S cell type.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229830 TI - Whole-cell K+ currents in fresh and cultured cells of the human and monkey retinal pigment epithelium. AB - 1. Whole-cell potassium currents of freshly isolated human (adult and fetal) and monkey (adult) retinal pigment epithelial (RPE) cells, as well as cultured human and monkey RPE cells were studied using the patch-clamp technique. 2. In freshly isolated adult cells of both species, two currents were observed in the voltage range from -150 to +50 mV: an outwardly rectifying current and an inwardly rectifying current. These currents were also found in cultured cells of both species. 3. The outwardly rectifying current in freshly isolated adult human and monkey cells and some cultured cells was evoked by depolarizing voltage pulses more positive that -30 mV. The current activated with a sigmoidal time course after a brief delay, and was virtually non-inactivating. The conductance associated with the current was half-maximal at -16.4 mV for fresh human cells and -13.5 mV for fresh monkey cells, but was shifted 16.0 and 17.7 mV in the positive direction in cultured human and monkey cells, respectively. The reversal potential of the current in both human and monkey cells matched the potassium equilibrium potential (EK) over a wide range of external potassium concentrations. This current was blocked by 20 mM tetraethylammonium. 4. A membrane current that exhibited inward rectification was observed with hyperpolarizing voltage pulses. The zero-current potential of this current was close to EK. This current was blocked by 2 mM Ba2+ and 2 mM Cs+. In cultured human and monkey cells, but not in fresh cells, this current exhibited an inactivation when voltage pulses were more negative than -120 mV. External Na+ was responsible for the inactivation, as the inactivation was removed in a Na(+) free solution. 5. Membrane currents in freshly isolated fetal human RPE cells were remarkably different from those in adult cells. A transient outward current resembling the A-type potassium current was observed as the dominant membrane current in freshly isolated fetal human cells. This current activated when voltage pulses were more positive than -30 mV. It inactivated rapidly after reaching a maximal level. Application of 5 mM 4-aminopyridine (4-AP) completely blocked this current. Although this current was never observed in fresh adult cells, it was found in 33% of the cultured adult cells with similar kinetics, ion selectivity, and pharmacological properties. 6. In about 26% of the freshly isolated fetal human cells, a more slowly activating outward current, which resembled the delayed rectifier, was found to co-exist with the transient outward current.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229831 TI - Chloride currents activated by caffeine in rat intestinal smooth muscle cells. AB - 1. Current responses to caffeine in single smooth muscle cells isolated from rat intestine were studied with the whole-cell patch clamp technique. Intracellular calcium concentration, [Ca2+]i, was simultaneously monitored with fura-2 (0.1 mM) introduced into the cell through a patch pipette. 2. With a potassium-containing pipette solution, caffeine (10 mM) produced an outward current at a holding potential of 0 mV and an inward current at -60 mV, both of which were accompanied by parallel increases in [Ca2+]i. The outward current response disappeared after the removal of K+ from pipette solutions, indicating that caffeine activates a Ca(2+)-activated K+ conductance. 3. When NaCl was present in both pipette and external solutions as the major constituent, caffeine evoked an inward current at -60 mV simultaneously with a rise in [Ca2+]i. The reversal potential (Er) of this current was about 0 mV. 4. Substitution of Tris+ or choline+ for external Na+ did not alter the Er. When external Cl- was replaced by thiocyanate-, iodide- or glutamate-, the Er changed to respectively -55, -38 and +35 mV. 5. The current response to caffeine decreased with increasing concentration of EGTA in the pipette solution. The caffeine-induced current and the intracellular Ca2+ transient was still observed for a few minutes after exposure of the cells to Ca(2+)-free external solution containing 2 mM EGTA. Caffeine failed to produce an inward current and Ca2+ transient after treatment with extracellular ryanodine. 6. It is concluded that caffeine caused an increase in membrane Cl- conductance and in K+ conductance resulting from a rise in [Ca2+]i derived from ryanodine sensitive intracellular Ca2+ stores in isolated smooth muscle cells of the rat intestine. PMID- 8229832 TI - ATP-sensitive K+ channel modification by metabolic inhibition in isolated guinea pig ventricular myocytes. AB - 1. ATP-sensitive K+ (K+ATP) channels are believed to make an important contribution to the increased cellular K+ efflux and shortening of the action potential duration (APD) during metabolic inhibition, hypoxia, and ischaemia in the heart. The mechanisms by which the activity of the K+ATP channel is regulated during conditions of metabolic impairment are not completely clear. Extrinsic factors such as increased [ADP]i, acidosis, and stimulation of adenosine receptors appear to decrease the K+ATP channel's sensitivity to closure by [ATP]i. The purpose of this study was to determine whether the K+ATP channel itself is intrinsically altered by the processes associated with metabolic impairment. 2. Isolated guinea-pig ventricular myocytes were metabolically inhibited in glucose-free 1.8 mM Ca2+ Tyrode solution containing 9 microM rotenone and 0.9 microM carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) while recording unitary currents through K+ATP channels in cell-attached patches. When K+ATP channel activity became maximal, the patch was excised (inside-out) into 150 mM K+ bath solution containing different ATP concentrations. The Kd for suppression by [ATP]i ([ATP]i causing half-maximal suppression of current through K+ATP channels) was markedly increased to 305 microM (n = 9) compared to patches excised from control myocytes not exposed to metabolic inhibitors (Kd = 46 microM, n = 28). 3. A [Ca2+]i-dependent process was involved in K+ATP channel modification during metabolic inhibition. Removal of extracellular Ca2+ during metabolic inhibition led to an intermediate decrease in the ATP sensitivity of the K+ATP channels (Kd = 120 microM, n = 6). In myocytes that were pretreated with 10 microM ryanodine in addition to removing extracellular Ca2+, the reduction in ATP sensitivity was completely prevented (Kd = 23 microM, n = 6). 4. In inside-out membrane patches excised from control non-metabolically inhibited myocytes, elevated free [Ca2+]i (2 microM) did not alter the sensitivity of the K+ATP channel to closure by [ATP]i, suggesting that in metabolically inhibited myocytes elevated [Ca2+]i acted indirectly. K+ATP channel run-down was found to increase the sensitivity of K+ATP channels to closure to [ATP]i (Kd = 16 microM, n = 13). 5. Inside-out membrane patches excised from control non-metabolically inhibited myocytes were also exposed to various proteases, phospholipases and other reagents that may be activated during metabolic inhibition. Trypsin and chymotrypsin treatment increased the Kd from 39 to 213 microM (n = 8) and 110 microM (n = 5), respectively. Calpain I had no apparent effect on the Kd.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229833 TI - Ca2+ channel modulating effects of heparin in mammalian cardiac myocytes. AB - 1. The effect of heparin on L-type Ca2+ channels in rabbit, rat and guinea-pig cardiac myocytes was studied using the whole-cell patch clamp method. 2. Sodium salts of heparin uniformly suppressed the Ca2+ current, ICa, independent of their molecular weight, in the rat and guinea-pig ventricular and rabbit atrial myocytes. The suppression of ICa by heparin was dose dependent and reached its maximum, about 30%, around 10 microM. Heparin did not alter the voltage dependence or the steady-state inactivation properties of ICa. These effects were specific to heparin as another polysaccharide, dextran, failed to have any effect on ICa. 3. The suppressive effect of heparin was not diminished when [Ca2+]o was increased to 10 mM, or when Ba2+ was the charge carrier through the Ca2+ channel. 4. Spectrophotometric assays showed that heparin-induced changes in [Ca2+]o generally were too small to alter ICa significantly. 5. In myocytes buffered with 0.1 mM EGTA, the suppressive effect of heparin was more prominent on the inactivating than on the maintained component of ICa. 6. When extracellular Na+ was replaced by Cs+, the heparin suppressive effect was accompanied by a 10 mV shift of both the voltage dependence of activation and the steady-state inactivation parameters toward more negative potentials. 7. When both Mg2+ and Na+ were omitted from the bathing solutions, the suppressive effect of heparin was significantly enhanced such that almost 80% of the current was blocked. 8. In Cs(+)-based solutions 10 mM [Mg2+]o suppressed ICa by about 70% and heparin partially relieved this block. Heparin, however, did not counteract the Mg(2+) induced suppression of ICa in Na(+)-based solution. 9. Extracellularly applied heparin did not alter the isoprenaline-induced enhancement of ICa or interfere with the blocking effect of phorbol esters on ICa. 10. Heparin thus appears to interfere with the permeation of Ca2+ through the channel by a mechanism regulating the Ca(2+)-induced inactivation of the Ca2+ channel. Na+ and Mg2+ appear to alter the kinetics and the magnitude of the suppressive effect of heparin on the Ca2+ channel, suggesting an interaction of these cations with either the Ca2+ or the heparin-binding sites of the channel. PMID- 8229834 TI - Changes of intracellular [Ca2+] during refilling of sarcoplasmic reticulum in rat ventricular and vascular smooth muscle. AB - 1. Intracellular calcium concentration ([Ca2+]i) was measured in single myocytes isolated from either the cardiac ventricle or the mesenteric artery of the rat. 2. In both cardiac and smooth muscle, the application of caffeine produced an increase of [Ca2+]i which spontaneously decayed back to resting levels. In vascular smooth muscle cells, removal of caffeine produced a transient fall of [Ca2+]i to below the resting level. [Ca2+]i then returned to control levels. A transient undershoot of [Ca2+]i on removal of caffeine was also sometimes seen in cardiac cells. When the undershoot was absent in cardiac cells it could be induced by elevating [Ca2+]o. 3. In vascular smooth muscle cells noradrenaline increased [Ca2+]i and an undershoot of [Ca2+]i could be produced by its removal. In cardiac cells a small undershoot could sometimes be seen following the systolic Ca2+ transient produced by electrical stimulation. 4. In both cardiac and vascular cells the time constant of decay of the caffeine response (tau caff) was less than that of the recovery from the undershoot (tau us). On average the ratio tau us:tau caff was about 5 in smooth muscle. In cardiac cells the recovery of the undershoot was also considerably slower than that of the caffeine response. 5. If caffeine was removed before the rise of [Ca2+]i had fully decayed spontaneously then the magnitude of the undershoot was reduced. 6. It is suggested that the undershoot of [Ca2+]i on removal of caffeine results from refilling of the SR decreasing [Ca2+]i. The data from vascular cells can be fitted by this model if the dissociation constant, Kd, of the surface membrane Ca2+ pump for [Ca2+]i is about 1 microM. 7. Using the model, it is concluded from the ratio of the time constants shown above that the caffeine releasable content of the sarcoplasmic reticulum constitutes about 80% of total cellular calcium in both cardiac and smooth muscle. PMID- 8229835 TI - Quantification of exponential Na+ current activation in N-bromoacetamide-treated cardiac myocytes of guinea-pig. AB - 1. The activation kinetics of the Na+ current was investigated in single ventricular cells of the guinea-pig heart using an improved oil-gap voltage clamp method. The inactivation of the current was removed by an intracellular application of N-bromoacetamide (NBA) for less than 1 min. Although the NBA treatment slightly decreased the peak amplitudes (81.7 +/- 13.4% of control, n = 15), the Na+ current remained stable after the removal of inactivation. 2. On depolarization, the activation of Na+ current took an exponential time course after the capacitive current decreased to 5% of its peak amplitude (40-100 microseconds after the pulse onset). The time course of deactivation, recorded on repolarization from 1.2 ms depolarization, was also a single exponential. 3. The time constants of activation and deactivation were almost identical when compared at a given test potential within a range of -50 to -30 mV. These findings indicate that the cardiac Na+ current activation is determined by m1 kinetics, or one rate-limiting step. 4. At potentials negative to -60 mV, the deactivation was complete, and its time constant decreased e-fold per 20.3 +/- 1.8 mV hyperpolarization (n = 7). 5. The degree of steady-state activation (m(infinity)) was fitted to a Boltzmann equation with a slope factor of 7.4 +/- 0.3 mV and a half-maximum potential of -33.3 +/- 0.8 mV (n = 8). 6. Rate constants for the rate-limiting activation step between a closed state and an open state (alpha m, beta m), were determined from m(infinity) and tau m over a potential range between -100 and +50 mV. On a logarithmic scale, beta m-1 was a linear function of the membrane potential over the range -100 and -30 mV. 7. Fitting the newly determined activation kinetics to the rising phase of the action potential indicated that the activation kinetics in the present study is relevant to the physiological action potential. The density of the Na+ channels thus obtained was 1075 +/- 186 pF-1 (n = 6). 8. The measurements in the NBA-treated Na+ current were compared with those obtained without treatment. PMID- 8229836 TI - Large conductance Ca(2+)-activated K+ channels are involved in both spike shaping and firing regulation in Helix neurones. AB - 1. The role of BK-type calcium-dependent K+ channels (K+Ca) in cell firing regulation was evaluated by performing whole-cell voltage clamp and patch clamp experiments on the U cell neurones in the snail Helix pomatia. These cells were selected because most of the repolarizing K+ current flowed through K+Ca channels. 2. U cells generated overshooting Ca(2+)-dependent spikes in Na(+)-free saline. In response to prolonged depolarizing current, they fired a limited number of spikes of decreasing amplitude, and behaved like fast-adapting or phasic neurones. 3. Under voltage clamp conditions, the K+Ca current had a slow onset at voltages that induced small Ca2+ entries. By manipulating the Ca2+ entry (either with appropriate voltage programmes or by changing the Ca2+ content of the bath), the K+Ca channel opening was found to be rate limited by the Ca2+ binding step and not by the voltage-dependent conformational change to the open state. 4. Despite the slow activation rate observed in voltage-clamped cells, 25 30% of the available K+Ca current was found to be active during isolated spikes. These data were based on patch clamp, spike-like voltage clamp and hybrid current clamp-voltage clamp experiments. 5. The fact that spikes led the slowly rising K+Ca current to shift into a fast activating mode was accounted for by the large surge of Ca2+ current concomitant with spike upstroke. The early calcium surge resulted in local increases in cytosolic calcium, which speeded up the binding of calcium ions to the closed K+Ca channels. From changes in the null Ca2+ current voltage, it was calculated that the submembrane [Ca2+]i increase to 50-80 microM during the spike. 6. Due to their fast voltage dependence, K+Ca channels appeared to play no role in shaping the interspike trajectory. 7. Even in the fast activating mode, the K+Ca current had a finite rate of rise and was not involved in repolarizing short duration Na(+-dependent action potentials. The current became more and more active, however, when voltage-gated K+ channels were progressively inactivated during firing. 8. The fast adaptation exhibited by U cells upon sustained depolarization was not paralleled by a recruitment of K+Ca channels because of the cumulative Ca2+ entries. During a spike burst, the K+Ca current progressively overlapped the depolarizing Ca2+ current, which ultimately stopped the firing. The early opening of K+Ca channels was ascribed to residual Ca2+ accumulation that kept part of the channels in the Ca(2+)-bound state ready to be opened quickly by cell depolarization.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229837 TI - Inward sodium current involvement in regenerative bursting activity of rat magnocellular supraoptic neurones in vitro. AB - 1. The rat hypothalamic slice preparation was used to investigate the involvement of inward Na+ currents as well as inward Ca2+ currents in the generation of bursting activity by supraoptic (SON) neurones. Intracellular records were made from thirty-two SON neurones which showed regenerative bursting activity. The bursting activity consisted of spontaneous, intermittent bursts of action potentials with subsequent silent periods. During the bursts, plateau potentials on which action potentials were superimposed were frequently observed. 2. Perfusion of a low-Na+ medium, a tetrodotoxin (TTX)-containing medium or a Ca(2+) free medium suppressed the regenerative bursting activity. 3. Addition of 3-10 microM veratridine to Ca(2+)-free medium elicited regenerative bursting activity and spontaneous plateau potentials. The veratridine-induced regenerative bursting activity and plateau potentials were blocked by 1 microM TTX. Addition of 5 mM TEA allowed regenerative bursting activity to persist in Ca(2+)-free medium. 4. These results suggest that TTX-sensitive Na+ inward currents as well as Ca2+ inward currents contribute to the generation of bursting activity in rat SON cells. PMID- 8229838 TI - Enhanced expression of Ca2+ channels by nerve growth factor and the v-src oncogene in rat phaeochromocytoma cells. AB - 1. Rat phaeochromocytoma (PC12) cells were used to investigate the expression of Ca2+ channel types during neuronal differentiation. Neuronal differentiation was induced by treatment with nerve growth factor (NGF) or by activation of a temperature-sensitive tyrosine kinase (pp60v-src) in genetically modified PC12 (PC12/v-src) cells. PC12 cells differentiated morphologically in the presence of NGF. When grown at the permissive temperature of 37 degrees C which activates the kinase activity of pp60v-src, PC12/v-src cells differentiated morphologically with the extension of neurites. In contrast, PC12/v-src cells grown at the non permissive temperature of 40 degrees C continued to divide and were morphologically indistinguishable from control PC12 cells. 2. Whole-cell Ca2+ currents were measured in PC12 cells using Ba2+ as the charge carrier. Ba2+ currents measured at the peak of the current-voltage curve from a holding potential of -80 mV were -0.28 +/- 0.04 nA (mean +/- S.E.M.) in control PC12 cells compared to -1.25 +/- 0.16 nA in NGF-differentiated cells. The current density increased from 9.4 +/- 0.7 pA/pF in control PC12 cells to 22.8 +/- 2.4 pA/pF in NGF-differentiated PC12 cells. Ba2+ currents were -0.24 +/- 0.04 nA in undifferentiated PC12/v-src cells grown at the non-permissive temperature of 40 degrees C compared to -0.95 +/- 0.16 nA in differentiated PC12/v-src cells grown at the permissive temperature of 37 degrees C. The current density increased from 4.5 +/- 0.5 pA/pF in PC12/v-src cells grown at the non-permissive temperature of 40 degrees C to 13.3 +/- 2.4 pA/pF in PC12/v-src cells grown at the permissive temperature of 37 degrees C. 3. The sensitivity of Ba2+ currents to omega conotoxin GVIA (omega-CgTX) was determined for currents measured at the peak of the current-voltage curve (0 mV in 10 mM Ba2+) from a holding potential of -80 mV. In NGF-differentiated PC12 cells, 10 microM omega-CgTx inhibited 68.1 +/- 3.2% of the total Ba2+ current compared to 35.9 +/- 4.1% in control cells. The density of the omega-CgTX-sensitive current increased from 3.3 +/- 0.4 pA/pF in control cells to 15.7 +/- 2.0 pA/pF in NGF-differentiated cells. In differentiated PC12/v-src cells grown at 37 degrees C, omega-CgTX inhibited 52.2 +/- 4.2% of total Ba2+ current compared to 41.1 +/- 3.8% in PC12/v-src cells grown at 40 degrees C. The density of the omega-CgTX-sensitive current increased from 1.9 +/- 0.3 to 7.4 +/- 2.0 pA/pF with v-src-mediated differentiation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229839 TI - Roles of sympathetic nervous system in the suppression of cytotoxicity of splenic natural killer cells in the rat. AB - 1. We previously demonstrated that a central injection of interferon-alpha in rats induced a suppression of cytotoxicity of splenic natural killer cells which depended upon intact splenic sympathetic innervation, suggesting the important role of the splenic nerve in immunosuppression. To further study the mechanisms of this phenomenon, we investigated: (1) the effects of a central injection of recombinant human interferon-alpha on the electrical activity of the splenic nerve, and (2) the responses of splenic natural killer cytotoxicity on the electrical stimulation of the splenic nerve in urethane with alpha-chloralose anaesthetized rats. 2. An injection of recombinant human interferon-alpha (1.5 x 10(3) and 6.0 x 10(3) units (u) per rat) into the third cerebral ventricle produced a sustained and long lasting (at least for more than 60 min) increase in the electrical activity of splenic sympathetic nerve filaments in a dose dependent manner. Following an intra-third-ventricular injection of recombinant human interferon-alpha at a dose of 6.0 x 10(3) u, the efferent discharges were elevated 2-6 times that of the pre-injection level with a mean onset latency of 12 min (8-16 min). No changes in the arterial blood pressure and body temperature were observed after injections of recombinant human interferon-alpha. 3. The excitation of the nerve activity induced by intra-ventricular recombinant human interferon-alpha was reversibly suppressed by an intravenous injection of an opioid antagonist, naloxone (1 mg/kg in 0.1 ml saline), whereas the injection of naloxone alone did not affect either the baseline level of the nerve activity or the systemic blood pressure. 4. The cytotoxicity of natural killer cells in the spleen measured by a standard chromium release assay was reduced 20 min after the laparotomy alone in anaesthetized rats. The reduced natural killer activity then recovered significantly when the splenic nerve was cut immediately after the laparotomy. When the peripheral cut end of the splenic nerve was subsequently stimulated (0.5 mA, 0.5 ms, 20 Hz for 20 min), a further suppression of natural killer cytotoxicity was observed. 5. The reduction of natural killer cytotoxicity produced by the stimulation of the splenic nerve was completely blocked by an intravenous injection of nadolol (a peripherally acting beta-adrenergic receptor antagonist), but not by that of prazosin (an alpha-antagonist). 6. These results indicate that a central injection of recombinant human interferon-alpha activates the splenic sympathetic nerve through brain opioid receptors and thereby suppresses the natural killer cytotoxicity by beta-adrenergic mechanisms.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229840 TI - Calcium release-activated calcium current in rat mast cells. AB - 1. Whole-cell patch clamp recordings of membrane currents and fura-2 measurements of free intracellular calcium concentration ([Ca2+]i) were used to study the biophysical properties of a calcium current activated by depletion of intracellular calcium stores in rat peritoneal mast cells. 2. Calcium influx through an inward calcium release-activated calcium current (ICRAC) was induced by three independent mechanisms that result in store depletion: intracellular infusion of inositol 1,4,5-trisphosphate (InsP3) or extracellular application of ionomycin (active depletion), and intracellular infusion of calcium chelators (ethylene glycol bis-N,N,N',N'-tetraacetic acid (EGTA) or 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA)) to prevent reuptake of leaked-out calcium into the stores (passive depletion). 3. The activation of ICRAC induced by active store depletion has a short delay (4-14 s) following intracellular infusion of InsP3 or extracellular application of ionomycin. It has a monoexponential time course with a time constant of 20-30 s and, depending on the complementary Ca2+ buffer, a mean normalized amplitude (at 0 mV) of 0.6 pA pF 1 (with EGTA) and 1.1 pA pF-1 (with BAPTA). 4. After full activation of ICRAC by InsP3 in the presence of EGTA (10 mM), hyperpolarizing pulses to -100 mV induced an instantaneous inward current that decayed by 64% within 50 ms. This inactivation is probably mediated by [Ca2+]i, since the decrease of inward current in the presence of the fast Ca2+ buffer BAPTA (10 mM) was only 30%. 5. The amplitude of ICRAC was dependent on the extracellular Ca2+ concentration with an apparent dissociation constant (KD) of 3.3 mM. Inward currents were nonsaturating up to -200 mV. 6. The selectivity of ICRAC for Ca2+ was assessed by using fura-2 as the dominant intracellular buffer (at a concentration of 2 mM) and relating the absolute changes in the calcium-sensitive fluorescence (390 nm excitation) with the calcium current integral. This relationship was almost identical to the one determined for Ca2+ influx through voltage-activated calcium currents in chromaffin cells, suggesting a similar selectivity. Replacing Na+ and K+ by N-methyl-D-glucamine (with Ca2+ ions as exclusive charge carriers) reduced the amplitude of ICRAC by only 9% further suggesting a high specificity for Ca2+ ions. 7. The current amplitude was not greatly affected by variations of external Mg2+ in the range of 0-12 mM. Even at 12 mM Mg2+ the current amplitude was reduced by only 23%. 8. ICRAC was dose-dependently inhibited by Cd2+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229841 TI - Volume-dependent K+ and Cl- fluxes in rat thymocytes. AB - 1. Hypotonic stress unmasked inward and outward K+ and Cl- movements in rat thymocytes. This KCl flux stimulation was reduced by DIOA (dihydroindenyl-oxy alkanoic acid), but not by DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonate), quinidine, DPAC 144 (5-nitro-2-(2-phenylethyl-amino)-benzoic acid), bumetanide or ouabain. 2. In isotonic media (308 +/- 5 mosmol kg-1), the cells exhibited the following DIOA-sensitive fluxes: (i) a K+ efflux of 42.7 +/- 17.1 mmol (l cells.h)-1 (mean +/- S.D., n = 7), (ii) a Cl- efflux of 68 +/- 21 mmol (l cells.h)-1 (n = 3), (iii) a Rb+ influx of 9.7 +/- 3.9 mmol (l cells.h)-1 (n = 6) and (iv) a Cl- influx of 9.4 +/- 4.1 mmol (l cells.h)-1 (n = 6). 3. Hypotonic shock (183-200 mosmol kg-1) induced a sevenfold stimulation of DIOA-sensitive K+ and Cl- effluxes and a twofold stimulation of DIOA-sensitive Rb+ and Cl- influxes (with a Rb+ to Cl- stoichiometry of 1.04 +/- 0.31; mean +/- S.D., n = 6). 4. The DIOA-sensitive membrane carrier catalysed net outward KCl extrusion (the outward/inward flux ratio was 5-7 in isotonic media and 20 in hypotonic media at 189 mosmol kg-1). Inhibition of DIOA-sensitive 36Cl- efflux by cell K+ depletion suggested coupling of outward K+ and Cl- fluxes. Conversely, inward K+ and Cl- fluxes were found to be uncoupled in NO3- media and in K(+)-free media. 5. The results clearly show that rat thymocyte membranes possess a 1:1 K(+)-Cl- co transport system which is strongly activated by hypotonic shock and catalyses net KCl extrusion. PMID- 8229842 TI - Cell volume regulation in rat thymocytes. AB - 1. DIOA (dihydroindenyl-oxy-alkanoic acid), a potent inhibitor of the K(+)-Cl- co transport system, fully blocked regulatory volume decrease (RVD) in swelled rat thymocytes, with an IC50 of 2.2 +/- 0.5 x 10(-5) mol l-1 (mean +/- S.D., n = 4). Conversely, RVD was resistant to quinine, quinidine, apamin, cetiedil, amiloride, bumetanide and DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonate). 2. DIOA sensitive RVD followed mono-exponential kinetics, with t1/2 (half-lifetime) of 1 3 min and maximal capacity (Cmax) of about 55% of the initial cell swelling. Cmax and the initial rate of RVD (Vo) were both linear functions of the increase in cell volume. 3. RVD was: (i) slightly increased by replacing external Cl- by NO3 , (ii) reversed by replacing external Na+ by K+ (in the presence of external Cl-) and (iii) inhibited by cell K+ depletion. All these phenomena were blocked by DIOA (86 mumol l-1). 4. Increased membrane potassium permeability by valinomycin was unable to accelerate RVD or RVD reversal. 5. In the presence of DIOA, thymocytes responded like osmometers (the relative cell volume was a linear function of the reciprocal of the relative osmolality) in a large range of osmolalities. 6. The results strongly suggest that RVD in rat thymocytes is mediated by the K(+)-Cl- co-transport system. PMID- 8229843 TI - A possible role for endogenous peripheral corticotrophin-releasing factor-41 in the febrile response of conscious rabbits. AB - 1. The actions of peripheral corticotrophin-releasing factor-41 (CRF-41) on the febrile responses of conscious rabbits induced by peripherally administered polyinosinic.polycytidylic acid (poly(I).poly(C)) have been studied using a CRF 41 receptor antagonist (alpha-helical CRF(9-41) and anti-CRF-41 monoclonal antibodies. 2. Temperature responses were monitored continuously using rectal thermistor probes. Test substances were administered intravenously (i.v.), or for central CRF-41 antagonism experiments, via an indwelling third ventricle cannula (I.C.V.). Blood samples were taken at time intervals from a marginal ear vein and plasma cortisol levels determined by radioimmunoassay. 3. Poly(I).poly(C) (2.5 micrograms/kg) stimulated a reproducible biphasic rise in body temperature with a lag phase of 45-60 min and peaks at 90 and 225 min. 4. The febrile response to poly(I).poly(C) (2.5 micrograms/kg I.V.) was antagonized by blockade of peripheral CRF-41 actions using either monoclonal anti-CRF-41 antibodies (2.5 mg/kg i.v.) or the CRF-41 receptor antagonist (alpha-helical CRF(9-41); 25 micrograms/kg i.v.) administered 5 min prior to the pyrogen. 5. Centrally administered CRF-41 receptor antagonist (2.5 micrograms/kg I.C.V.) failed to affect the febrile response to poly(I).poly(C) (2.5 micrograms/kg i.v.). 6. CRF 41 immunoneutralization after the onset of temperature rises caused an immediate and significant defervescence. 7. In conclusion, these results suggest a modulatory pro-pyretic role for endogenous peripheral CRF-41 in the febrile responses to poly(I).poly(C). PMID- 8229844 TI - Effects of metabolic inhibition and changes of intracellular pH on potassium permeability and contraction of rat uterus. AB - 1. We have investigated the role of changes of potassium efflux in the inhibition of uterine force produced by cyanide. K+ efflux (86Rb) was measured from pregnant and non-pregnant rat myometrial strips during metabolic inhibition with cyanide and following manoeuvres to displace intracellular pH (pHi). 2. Cyanide greatly reduced or abolished spontaneous contractions. If the membrane was depolarized directly at this stage (by elevating external K+) then contraction redeveloped. This suggests that the initial depression of force is due to a failure of membrane excitation. 3. Cyanide reversibly increased 86Rb efflux (30-35%) in both pregnant and nonpregnant uteri and contraction was reduced. The increase in 86Rb efflux with cyanide was not secondary to changes of membrane potential as it also occurred in both high-K+ and Ca(2+)-free solutions. 4. Glibenclamide (20 microM), an antagonist of K+ATP channels, reduced the cyanide-evoked increase of 86Rb efflux by about 50%. The glibenclamide-insensitive component of efflux persisted in a Ca(2+)-free solution. Despite its action on 86Rb efflux, glibenclamide did not restore contraction. 5. Intracellular pH falls during metabolic inhibition. We therefore investigated whether reducing pHi (in the absence of cyanide) had an effect on 86Rb efflux. Application of the weak acid butyrate (60 mM, at constant external pH, 7.4) had no significant effect on 86Rb efflux. Thus it is unlikely that the acidification in hypoxia contributes to the increased K+ efflux. 6. Intracellular alkalinization produced by the weak base trimethylamine (60 mM) increased the frequency of uterine contraction and the 86Rb efflux. However, there was no effect on the 86Rb efflux in a Ca(2+)-free solution. The increased efflux is therefore presumably a consequence of the increased frequency. 7. It is concluded that metabolic inhibition produced by cyanide, produces an increase in K+ efflux from the myometrium. Part of this efflux is glibenclamide sensitive. This increased K+ efflux will lead to hyperpolarization of the myometrial membrane and thus decrease excitation. Thus reduced surface membrane excitability will contribute to the fall of force in hypoxia; specifically it may cause the initial loss of spontaneous contractions in the uterus. PMID- 8229845 TI - Food stimulation of histidine decarboxylase messenger RNA abundance in rat gastric fundus. AB - 1. Histidine decarboxylase in the enterochromaffin-like cells of the gastric corpus mucosa converts histidine to histamine which in turn stimulates gastric acid secretion. The control of histidine decarboxylase activity is poorly understood. We have examined how fasting and refeeding influence the abundance of the messenger RNA encoding histidine decarboxylase in the gastric corpus of the rat. 2. The polymerase chain reaction was used to generate a probe for detection of histidine decarboxylase messenger RNA in Northern and slot blots of total RNA from the gastric corpus of rats fasted for up to 48 h, or fasted and then refed. A gastrin monoclonal antibody was used to neutralize the action of endogenous gastrin. 3. Fasting progressively reduced histidine decarboxylase messenger RNA abundance by 3- to 4-fold after 48 h. Refeeding induced a rapid increase in histidine decarboxylase messenger RNA abundance which was detectable after 30 min. 4. There was a significant correlation between histidine decarboxylase messenger RNA abundance and plasma gastrin. Administration of gastrin antibody inhibited the increase in histidine decarboxylase activity after 6 h refeeding, but not after refeeding for 30 min. 5. The results suggest that histamine mediated changes in postprandial acid secretion depend on control of histidine decarboxylase mRNA levels, and that gastrin regulates production of this enzyme in the rat over periods of a few hours. PMID- 8229846 TI - Influence of body position on pressure and airflow generation during hypoxia and hypercapnia in man. AB - 1. Inspiratory oesophageal pressure and ventilatory responses to hyperoxic, progressive hypercapnic rebreathing (HCVR) and isocapnic, progressive hypoxic rebreathing (HVR) were studied in five normal males in both supine and upright seated positions. 2. No significant differences were found in the ventilatory response to hypercapnia between the supine and upright position. The slopes of the relationship between minute ventilation (VI) and the increase of end tidal PCO2 (delta P(ET), CO2) were 3.27 +/- 0.23 and 2.76 +/- 0.24 1 min-1 mmHg-1 supine and upright, respectively. However, the change in oesophageal pressure from the end expiratory level observed during quiet breathing to that at peak inspiration (delta P(oes), I) in relationship to delta P(ET),CO2 was greater supine than upright (1.23 +/- 0.07 versus 0.79 +/- 0.11 cmH2O mmHg-1, P < 0.01). 3. In contrast, during hypoxia-stimulated breathing the slope of the minute ventilation versus oxyhaemoglobin saturation curve (VI-Sa,O2) was flatter supine than upright (1.00 +/- 0.03 versus 1.75 +/- 0.05 l min-1 (percentage fall in Sa,O2)-1, P < 0.0001), but delta P(oes), I in relation to Sa,O2 during hypoxic rebreathing was similar supine and upright (0.38 +/- 0.03 versus 0.40 +/- 0.04 cmH2O (percentage fall in Sa,O2)-1, respectively. 4. It is concluded that body position does not affect the ventilatory response to progressive hyperoxic hypercapnia but does affect the relationship between delta P(oes), I and delta P(ET),CO2. In contrast, body position affects the ventilatory response to isocapnic progressive hypoxia, but does not affect the relationship between delta P(oes), I and Sa,O2. PMID- 8229847 TI - Non-cholinergic nervous control of catecholamine secretion from perfused bovine adrenal glands. AB - 1. Field stimulation of adrenal nerves was used to study nervous control of adrenal catecholamine secretion in isolated, retrogradely perfused, bovine adrenal glands. 2. Secretion of both adrenaline and noradrenaline was maximal at 10 Hz. Secretion at 2 Hz was < 10% of maximum. Stimulating with trains of pulses at ten times the average frequency for 1 s out of every 10 s gave 2-fold greater secretion at 2 Hz average frequency, similar release at 5 Hz, and only half the secretion at 10 Hz, compared to continuous stimulation at the average frequency. 3. At 10 Hz, adrenaline and noradrenaline secretion was virtually abolished by tetrodotoxin (1 microM), but was only reduced by 75% by prolonged perfusion with a combination of mecamylamine (5 microM) and atropine (1 microM). Mecamylamine and atropine completely abolished the secretory response to 2 Hz stimulation. Tetrodotoxin had no significant effect on secretion induced by perfusing glands with nicotine (5 microM), while mecamylamine abolished this response. Mecamylamine and atropine had no effect on secretion induced by K+ depolarization. 4. The secretion of adrenaline and noradrenaline induced by 10 Hz stimulation was not inhibited by naloxone at either 1 or 30 microM. 5. The results suggest that bovine adrenal chromaffin cells, like those in the rat, receive a significant non-cholinergic secretomotor innervation. In contrast to the rat, however, the non-cholinergic component in the bovine adrenal is negligible at low-frequency nerve stimulation and substantial at higher frequencies, and is not antagonized by naloxone. The identity of the non cholinergic transmitter remains to be determined. PMID- 8229848 TI - Visual transduction in dialysed detached rod outer segments from lizard retina. AB - 1. Properties of a new preparation for studying the physiology and biochemistry of phototransduction in retinal rods are described. Whole-cell voltage clamp was used to record the generation, maintenance and light-sensitivity of dark current in rod outer segments that had been isolated from the rest of the receptor cell by detachment at the connecting cilium. 2. Detached outer segments dialysed with standard internal solution supplemented with physiological amounts of ATP (5 mM) and GTP (1 mM) developed a standing inward dark current that was the sum of three components: approximately 91% light-sensitive current, approximately 6% Na(+) Ca2+,K+ exchange current and approximately 3% leakage current. Light-sensitive dark current (mean amplitude approximately -63 pA) was suppressed transiently by brief flashes in an intensity-dependent manner. Light responses had the same kinetics, sensitivity and intensity-response relationship as those recorded from intact rods. 3. Dialysed outer segments differed from intact rods in that intense flashes evoked saturating responses that recovered incompletely to a plateau of reduced dark current caused by incomplete inactivation of the transduction cascade. Light sensitivity was reduced for a short time following an intense flash and then recovered despite persistent reduction of dark current. This suggests that there is no fixed relationship between dark current amplitude and light sensitivity. 4. Light-sensitive dark current faded rapidly when outer segments were not supplied with nucleotides. Outer segments dialysed with solution that contained cyclic GMP, but no ATP or GTP, supported dark current at a level that increased with [cyclic GMP]. When basal phosphodiesterase (PDE) activity is inhibited, 8 microM cyclic GMP supports a dark current of approximately 70 pA. 5. Light sensitivity decreased during recordings made with solution that contained only cyclic GMP, consistent with the inhibition of G protein activation by loss of GTP. After thorough nucleoside triphosphate depletion, however, intense illumination evoked a transient increase rather than a decrease in dark current, i.e. an inverted light response. This result suggests that isomerized rhodopsin may generate a signal that causes either inhibition of basal PDE activity or release of bound cyclic GMP. 6. Sustained Na(+)-Ca2+,K+ exchange current was recorded during steady illumination when Ca2+, but not when Mg2+, was added to the dialysis solution. Exchange current increased with the amount of added Ca2+ and saturated at approximately 18 pA when the dialysis solution contained > or = 10 mM Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229849 TI - Effect of extravascular plasma protein on pressure-flow relations across synovium in anaesthetized rabbits. AB - 1. The effect of extravascular plasma protein on fluid flux through interstitial matrix was investigated in vivo by studying the pressure-flow relation across synovium during intra-articular infusions of protein solutions (usually bovine serum albumin). Synovium is a sheet of non-epithelial cells separated by interstitium-filled gaps, beneath which are fenestrated capillaries: synovium regulates synovial fluid volume and composition. 2. Albumin solutions (10-150 g l 1) of measured oncotic pressure and viscosity were infused at known pressure into the synovial cavity of knees of anaesthetized rabbits. Flow across the synovial lining in the steady state (absorption rate Qs) was recorded at a series of joint pressures (Pj) to define the pressure-flow relation. Krebs solution was infused into the opposite knee as a control (26 animals). 3. Infusion of a low albumin concentration (10 g l-1, bovine or rabbit) or diluted rabbit serum revealed no specific effect of plasma protein on interstitial matrix permeability (cf. specific protein effect on capillary glycocalyx permeability). Physiological (22.5 g l-1) and higher concentrations reduced trans-synovial absorption rate. The slope of the pressure-flow relation was reduced and the pressure intercept displaced to the right (i.e. Pj at zero flow was raised). 4. Slope dQs/dPj correlated negatively with intra-articular viscosity (P = 0.001-0.04), in keeping with viscous interstitial flow. The reduction in normalized slope, however, did not equal the reduction in fluidity (1/viscosity) quantitatively. It is proposed that apparent fluidity within the interstitial matrix is higher than in the bulk phase due to steric exclusion of albumin (radius 3.55 nm) by the interstitial glycosaminoglycans. The latter form spaces of estimated mean hydraulic radius 14 18 nm in synovium. 5. The joint-pressure intercept at zero net trans-synovial flow was displaced 0.015 cmH2O per cmH2O intra-articular oncotic pressure (pi j; S.E.M. +/- 0.006). Thus large trans-synovial osmotic gradients were not maintained at physiological flow velocities. The 1.5% displacement of the Pj intercept by pi j was attributed principally to interstitial albumin exerting pericapillary oncotic pressure and enhancing net Starling filtration pressure. Indeed, net trans-synovial flow at low joint pressure sometimes reversed from absorption to filtration into the joint cavity at high intra-articular oncotic pressures. 6. The displacement of the trans-synovial flow intercept per unit change in intra-articular oncotic pressure, (dQs/d pi j)Pj = 0, was 18 +/- 3 nl min-1 cmH2O-1.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229850 TI - A relay for input from group II muscle afferents in sacral segments of the cat spinal cord. AB - 1. A neuronal relay for input from group II afferents of hindlimb muscle nerves has been found in the previously little explored sacral segments of the cat spinal cord. 2. Electrical stimulation of group II muscle afferents of a number of nerves evoked negative potentials on the surface (cord dorsum potentials) and population postsynaptic potentials (field potentials) within the sacral segments. The largest potentials were evoked by stimulation of the posterior biceps semitendinosus and triceps surae nerves which evoke much smaller potentials in other segments. Group II afferents of other nerves, notably those which have their main relay within the middle lumbar segments, were much less effective. 3. The sites at which cord dorsum and field potentials evoked by group II muscle afferents were recorded varied in relation to the external topography of the L7 S2 spinal segments but were consistent in their location relative to the pudendal motor nucleus (Onuf's nucleus). Potentials evoked by group II afferents of the posterior biceps and semitendinosus nerves peaked at a level corresponding to the rostral half of Onuf's nucleus and potentials evoked by afferents of the gastrocnemius nerves peaked just rostral to this nucleus. The largest field potentials (of 0.5-1.0 mV) were recorded within the dorsal horn. Field potentials in the intermediate zone were much smaller (< 0.3 mV) and were seen less frequently. 4. Evidence was obtained that the dorsal horn field potentials are to a great extent evoked monosynaptically by the fast conducting fraction of group II muscle afferents: (i) they were evoked at short latencies (2.4-2.7 ms from the stimulus; 1.3-1.7 ms from group I components of afferent volleys and 0.5-0.7 ms from group II components of these volleys), (ii) the conduction times of impulses in the fastest conducting fraction of group II afferents, between the sacral segments (where these impulses were induced by intraspinal stimuli) and the peripheral nerves, were only about 0.5 ms shorter than the latencies of field potentials recorded at the site of intraspinal stimulation and evoked by stimulation of the same peripheral nerves and, (iii) the field potentials followed repetitive stimuli without temporal facilitation. 5. Negative cord dorsum and field potentials were also evoked by small stretches of the semitendinosus and triceps surae muscles. Although they were smaller than potentials evoked by electrical stimulation of sensory fibres and appeared at longer latencies, their presence is consistent with a contribution of muscle spindle afferents to the actions of group II muscle afferents within the sacral segments.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229851 TI - Mechanisms of zinc transport into pig small intestine brush-border membrane vesicles. AB - 1. The purpose of the present work was to examine certain membrane transport mechanisms likely to carry zinc across the brush-border membrane of pig small intestine, isolated in a vesicular form. 2. In initial velocity conditions, saturation kinetics revealed a great effect of pH on zinc transport: optimal conditions were observed with an intravesicular pH of around 6.6 with or without a H+ gradient; however, this did not allow us to conclude the existence of a neutral exchange between Zn2+ and H+ ions. 3. By measuring 36Cl uptakes, the presence of the Cl(-)-HCO3- or Cl(-)-OH-antiporter with typical 4,4' diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) sensitivity was detected in vesicles; zinc did not alter this anionic exchange activity. A 65Zn time course, performed in conditions identical with those for 36Cl uptake, was DIDS insensitive and was greatly inhibited by an outward OH- gradient. This could argue against a transport of zinc as a complex with Cl- and HCO3- through the anion antiporter. 4. When external Cl- and HCO3- were replaced by SCN-, able to form a Zn(SCN)4(2-) complex, we observed a stimulating effect of outward HCO3- gradients on 65Zn uptake but neither DIDS nor diphenylamine-2-carboxylate (DPC) inhibited the transport in these conditions. This suggested that the intestinal anion antiporter was not a major route for zinc reabsorption. 5. The tripeptide Gly-Gly-His at low concentrations stimulated 65Zn uptake, then inhibited it in a dose-dependent manner either in the presence of an inward H+ gradient or in the presence of a membrane potential 'negative inside' or in both situations. These conditions are necessary for the active transport of the peptide and this strongly suggests that zinc can be transported as a [Gly-Gly-His-Zn] complex, utilizing the peptide carrier system. PMID- 8229852 TI - Role of periodontal mechanoreceptors in evoking reflexes in the jaw-closing muscles of the cat. AB - 1. In anaesthetized cats the left maxillary and mandibular canine and incisor teeth were extracted. Nine weeks later the animals were anaesthetized again and titanium implants were placed into the edentulous ridge of the maxilla and mandible. 2. Terminal experiments were performed between 9 weeks and 10 months after inserting the implants. Recordings were made from the trigeminal ganglia and peripheral nerves of anaesthetized cats. Neurones were identified which responded to forces applied to the maxillary teeth but none was found which responded to forces applied to the implant. 3. The responses of motor units in the temporalis muscle were studied in response to ramp-plateau forces applied to the maxillary canine tooth and maxillary implant using peristimulus time histograms and cumulative sum (cusum) analysis. A small amount of inhibition of the motor units was observed in response to the applied forces to the implant. However, the inhibition was much more profound when similar forces were applied to the tooth. 4. The results confirm that when forces are applied to a tooth, periodontal mechanoreceptors are stimulated which evoke reflex inhibitions to motor units in the jaw-closing muscles. However, there is evidence that mechanoreceptors situated distant to the periodontium can also evoke such reflexes. 5. The results are discussed in relation to the overall functional role of periodontal mechanoreceptors in the reflexes of mastication. PMID- 8229853 TI - Effect of route of breathing on the ventilatory and arousal responses to hypercapnia in awake and sleeping dogs. AB - 1. The influence of the upper airway on the ventilatory and arousal responses to hypercapnia in wakefulness and sleep was investigated using a chronic animal model. 2. Experiments were performed in five unrestrained dogs trained to sleep naturally in the laboratory. The animal rebreathed through a chronic tracheostoma (thus excluding the upper airway from the breathing circuit), or through the snout (intact upper airway). Resistance to breathing and volume of dead space during quiet tracheal breathing were matched to those in quiet nasal breathing during wakefulness and sleep. CO2 rebreathing tests were performed during wakefulness, rapid eye movement (REM) and non-REM (NREM) sleep, during nasal and tracheal breathing. 3. The ventilatory response to hypercapnia was significantly lower in nasal breathing compared with tracheal breathing, in all behavioural states. This was due to a smaller tidal volume and lower breathing frequency. 4. The ventilatory response to CO2 was lowest during REM sleep, irrespective of route used for breathing. 5. Alveolar partial pressure of CO2 (PA,CO2) level at arousal was identical in NREM nasal and tracheal rebreathing tests. Differences in PA,CO2 levels at arousal between NREM and REM sleep were not significant in nasal tests and only marginally different during tracheal breathing. 6. We conclude that nasal breathing influences the hypercapnic ventilatory response in wakefulness and sleep, and that the presence of CO2 in the upper airway does not affect arousal in NREM and REM sleep. PMID- 8229854 TI - Effects of pH and inorganic phosphate on force production in alpha-toxin permeabilized isolated rat uterine smooth muscle. AB - 1. Strips of longitudinal smooth muscle isolated from rat uterus were permeabilized using crude alpha-toxin from the bacterium Staphylococcus aureus. This treatment rendered the surface membrane permeable to small molecular weight substances. Simultaneous measurements of tension and calcium concentration ([Ca2+]) (using indo-1 fluorescence) were used to investigate the effects of pH and inorganic phosphate concentration ([Pi]) on Ca(2+)-activated force generated by the contractile proteins. 2. Raising the [Pi] from 1 to 11 mM at a pH of 7.2 depressed both maximal and submaximal Ca(2+)-activated force. This effect of Pi was concentration dependent having the majority of its effect by 6 mM. 3. Further experiments at a submaximal [Ca2+] showed that Ca(2+)-activated force was enhanced by raising [Pi] from 6 to 11 mM suggesting that Pi increased the Ca2+ sensitivity of tension production. Based on these results, calculations indicate that the apparent affinity constant of Ca2+ for the contractile proteins increased from 4 x 10(6) M-1 to 6 x 10(6) M-1 on raising [Pi] from 1 to 11 mM. 4. Lowering pH from 7.2 to 6.7 at a [Pi] of 1 mM potentiated Ca(2+)-activated force with a small depression in the apparent Ca2+ sensitivity of tension production. This effect of pH on maximum (100 microM Ca2+) and submaximum (0.3 microM Ca2+) Ca(2+)-activated force was observed over a range of acidic pHs (7.0-6.7). 5. Increasing pH from 7.2 to 7.7 at a [Pi] of 1 mM depressed Ca(2+)-activated force with no effect on Ca2+ sensitivity of tension production. 6. Spontaneous contractions in intact rat myometrium are abolished under hypoxic conditions. Under these same conditions intracellular [Pi] rises and pH falls. The results of this study suggest that taken individually neither the effect of a rise in [Pi] nor a fall in pH on Ca(2+)-activated force generated by the contractile proteins can account for the effect of hypoxia on spontaneous contractions. PMID- 8229855 TI - Projection of cat jaw muscle spindle afferents related to intrafusal fibre influence. AB - 1. A method of classification of muscle spindle afferents using succinylcholine (SCh) and ramp stretches has recently been described, which appears to estimate separately the strength of influence of bag1 (b1) and of bag2 (b2) intrafusal fibres. Increase in dynamic difference (delta DD) indicates b1 influence whilst increase in initial frequency (delta IF) indicates b2 influence. The significance of this classification has now been examined by correlation with the strength of synaptic projection of jaw muscle spindle afferents to the fifth motor nucleus (MotV) and the supratrigeminal region (STR) in anaesthetized cats. 2. Projection strength was estimated by computing the extracellular focal synaptic potential (FSP) from spike-triggered averages of 1024 sweeps at 100 microns intervals along tracks through STR and MotV. Trigger pulses were derived from spindle afferent cell bodies of the jaw-closer muscles recorded in the mesencephalic trigeminal nucleus, and characterized by the effect of SCh on their responses to ramp-and hold stretches. 3. The maximum size of FSPs in tracks traversing STR and MotV ranged from 2.08 to 36.99 microV with a mean of 7.55 microV. The amplitudes were bimodally distributed into roughly equal-sized groups with high and low amplitude FSPs. 4. Mean values of delta IF were significantly greater for the group with large FSPs than for those with small FSPs. There were no significant differences in delta DD. FSP amplitude was significantly positively correlated with delta IF, but not with delta DD. 5. Spindle afferents with high values of FSP amplitude in MotV had a wide range of values of delta DD (b1b2c and b2c groups), while units with large FSPs in STR were all in the b2c category. Some evidence is presented to indicate that this reflects a preferential projection of secondary afferents to the STR. 6. For those units with projection to both STR and to MotV, there was a significant positive correlation between FSP amplitude in the two nuclei. 7. These results indicate that the extent of the b2 influence on spindle afferents predicts the central projection strength better than does the b1 influence. This finding is discussed from the viewpoint of possible developmental and functional issues. PMID- 8229856 TI - Histamine activates Cl- and K+ currents in guinea-pig tracheal myocytes: convergence with muscarinic signalling pathway. AB - 1. We investigated the effects of histamine on membrane currents and contractile state of isolated guinea-pig tracheal myocytes using perforated patch and whole cell recording techniques. The effects of histamine were compared to those of acetylcholine (ACh) and caffeine. 2. During voltage clamp (Vhold = -60 mV), histamine elicited contraction and an inward current (Ihist) which was often followed by current oscillations. Ihist had a reversal potential (Vrev) of -9 +/- 3 mV. 3. Ihist was dependent on the Cl- gradient and was antagonized by the Cl- channel blocker niflumic acid. Vrev was more positive (+2 +/- 1 mV) when K(+) selective currents were blocked by Cs+ and TEA. When all external Na+ was replaced with N-methyl-D-glucamine, there was a small reduction in the amplitude of Ihist. 4. The histamine-induced current was similar to that elicited by ACh and by caffeine with respect to time course, amplitude, and current-voltage relationship. Responses to histamine and to ACh were non-additive, consistent with a convergence of histaminergic and cholinergic signalling pathways. Ihist was antagonized by the H1 histaminergic receptor antagonist astemizole, but not by atropine. 5. When recorded using the perforated patch configuration, Ihist could be elicited repeatedly for more than 30 min. When cells were studied in the whole-cell configuration using a pipette solution containing 0.025 mM EGTA, the amplitude of Ihist was initially the same as that obtained using perforated patch but then decreased; the time required for the responses to decrease to 50% (t1/2) was 8.2 +/- 1.0 min. When 1 mM EGTA was included in the pipette solution (whole cell configuration), the initial response to histamine was significantly decreased in size and t1/2 was reduced to 3.3 +/- 0.7 min. 6. The characteristics of the signalling pathway were examined in cells studied using the whole-cell configuration with 0.025 mM EGTA in the recording pipette. Heparin significantly reduced t1/2 to 4.3 +/- 0.8 min. GTP gamma S elicited inward current and oscillations; both effects were enhanced by histamine. GTP gamma S also reduced t1/2 to 1.4 +/- 0.1 min. Pertussis toxin did not alter the amplitude or time course of Ihist. 7. We conclude that in guinea-pig tracheal myocytes, binding of histamine to H1 receptors leads to release of Ca2+ from intracellular stores and subsequent activation of Cl- and K+ conductances as well as contraction. Furthermore, we demonstrate that ACh elicits similar physiological responses due to a convergence of the histaminergic and muscarinic signalling pathways. PMID- 8229857 TI - Hyperglycaemic hypoxia alters after-potential and fast K+ conductance of rat axons by cytoplasmic acidification. AB - 1. The effects of hyperglycaemic hypoxia (a condition possibly involved in the pathogenesis of diabetic neuropathy) on the depolarizing after-potential and the potassium conductance of myelinated rat spinal root axons were investigated using electrophysiological recordings from intact spinal roots and from excised, inside out axonal membrane patches. 2. Isolated spinal roots were exposed to hypoxia in solutions containing normal or high glucose concentrations. The depolarizing after-potential of compound action potentials was only enhanced in spinal roots exposed to hyperglycaemic (25 mM D-glucose) hypoxia. A maximal effect was seen in bathing solutions with low buffering power. 3. The depolarizing after-potential was also enhanced by cytoplasmic acidification after replacement of 10-30 mM chloride in the bathing solution by propionate. 4. Multi-channel current recordings from excised, inside-out axonal membrane patches were used to study the effects of cytoplasmic acidification on voltage-dependent K+ conductances with fast (F channels) and intermediate (I channels) kinetics of deactivation. 5. F channels were blocked by small changes in cytoplasmic pH (50% inhibition at pH 6.9). I channels were much less sensitive to intra-axonal acidification. 6. In conclusion, our data show that hyperglycaemic hypoxia enhances the depolarizing after-potential in peripheral rat axons. The underlying mechanism seems to be an inhibition of a fast, voltage-dependent axonal K+ conductance by cytoplasmic acidification. This alteration in membrane conductance may contribute to positive symptoms in diabetic neuropathy. PMID- 8229858 TI - Na+ pump current-voltage relationships of rabbit cardiac Purkinje cells in Na(+) free solution. AB - 1. The Na+ pump current (Ip) of isolated, single rabbit cardiac Purkinje cells in Na(+)-free solution was measured at 32-34 degrees C by means of whole-cell recording. 2. The Ip amplitude was studied as a function of clamp potential (Vc) and external concentration of various monovalent cations known to activate the Na(+)-K+ pump. 3. Under conditions which strongly activated Ip the Ip-Vc curve of the cells displayed a positive slope at membrane potentials negative to -20 mV and little variation at more positive potentials. 4. The Ip-Vc relationship showed an extended region of negative slope at positive and negative potentials in solutions containing low concentrations of activator cations which caused little Ip activation. A positive slope of the Ip-Vc curve was occasionally observed at clamp potentials negative to -60 mV under these conditions. 5. The shape of the Ip-Vc relation was independent of the cation species used as external Ip activator. 6. At zero membrane potential half-maximum Ip activation (K0.5(Vc = 0 mV) occurred at 0.05 mM Tl+, 0.08 mM K+, 0.4 mM NH4+ and 1.5 mM Cs+. The Hill coefficient derived amounted to 0.9 for Tl+, 1.2 for K+, 1.04 for NH4+ and 1.5 for Cs+. 7. The concentrations of external activator cations required for half-maximum Ip activation increased with depolarization. The voltage dependence of the K0.5 values could be described by a single exponential function for clamp potentials positive to -40 mV. 8. The steepness of the function is determined by a factor alpha, indicating the apparent fraction of an elementary charge which moves in the electrical field across the sarcolemma when external monovalent cations bind to the Na(+)-K+ pump. 9. The alpha values were calculated to be 0.32 for Tl+, 0.24 for K+, 0.29 for NH4+ and 0.18 for Cs+. Possible interpretations of the alpha values are considered. 10. It is suggested that binding of external monovalent activator cations to the Na(+)-K+ pump (or a process related to the binding) is voltage dependent. This potential-dependent process determines mainly the shape of the Ip-Vc curve in cardiac Purkinje cells superfused with Na(+)-free media containing low concentrations (< K0.5(Vc = 0 mV)) of K+ or its congeners. PMID- 8229859 TI - Interaction between central pattern generators for breathing and swallowing in the cat. AB - 1. We examined the interaction between central pattern generators for respiration and deglutition in decerebrate, vagotomized, paralysed and ventilated cats (n = 10), by recording activity from the following nerves: hypoglossal, phrenic, thyroarytenoid and triangularis sterni. Fictive breathing was spontaneous with carbon dioxide above the apnoeic threshold (end-tidal PCO2, 32 +/- 4 mmHg) and fictive swallowing was induced by stimulating the internal branch of the left superior laryngeal nerve (SLN) continuously (0.2 ms pulse duration, 10 Hz). 2. In all ten animals, SLN stimulation evoked short bursts of thyroarytenoid and hypoglossal nerve activity indicative of fictive swallowing. In two of ten animals, respiration was inhibited completely during deglutition. In the other eight animals, fictive breathing and swallowing occurred simultaneously. 3. With SLN stimulation below threshold for eliciting swallowing, the respiratory rhythm decreased, the duration of inspiration did not change but the duration of expiration, especially stage II, increased. Integrated nerve activities indicated that the rate of rise and peak of phrenic nerve activity decreased, stage I expiratory activity of the thyroarytenoid and especially that of the hypoglossal nerve increased and stage II expiratory activity of the triangularis sterni nerve was suppressed completely. However, if inspired carbon dioxide was increased, i.e. hypercapnic ventilation, stage II expiratory activity remained partially during continuous SLN stimulation. 4. Fictive-swallowing bursts occurred only at respiratory phase transitions. At the minimal stimulus intensity that evoked repetitive swallowing bursts, the pattern of interaction between breathing and swallowing central pattern generators was consistent for each animal (n = 7) but was different across animals. In four animals, fictive swallows occurred at the phase transition between stage II expiration and inspiration, at the transition between inspiration and stage I expiration in one animal; and in two other animals, at the transition between stage I and II of expiration. 5. The response to SLN stimulation accommodated during the stimulus train. Accommodation was evident in both the interswallow interval (ISI) which lengthened, and the interaction pattern which had fewer swallows per breath as the stimulus period progressed. In contrast to the ISI, characteristics of the fictive swallow did not accommodate. For example, duration of the swallow was constant, distributed over a narrow range throughout the stimulus train. 6. We conclude that the central pattern generators for swallowing and breathing interact. The pattern of interaction supports the three-phase theory of respiratory pattern generation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229860 TI - Intracellular sodium activity and its regulation in guinea-pig atrial myocardium. AB - 1. Intracellular Na+ activity (aNai) and membrane resting potential were studied in quiescent guinea-pig atrial and papillary muscles by means of Na(+)-sensitive and conventional microelectrodes. The effects of the cardioactive steroid dihydroouabain (DHO) on aiNa, force of contraction and sarcolemmal Na+, K(+) ATPase activity were also investigated. 2. In thirty atria and twenty-two papillary muscles, aNai amounted to 8.0 +/- 0.2 and 4.7 +/- 0.3 mM, respectively (mean +/- S.E.M.). When both tissues were from the same animal, with the same ion sensitive microelectrode mean aNai values of 7.9 +/- 0.2 and 5.1 +/- 0.5 mM (P < 0.01) were obtained from eight atrial and eight papillary muscles, respectively. 3. Membrane resting potentials (Em) were significantly (P < 0.001) more negative in the papillary muscles (-83.5 +/- 0.7 mV; n = 8) than in the atrium (-78.1 +/- 0.5 mV; n = 8). Deviation of Em from EK (determined by K(+)-sensitive microelectrodes) was 3.0 +/- 0.2 mV in ventricular (P < 0.05) and 6.1 +/- 0.3 mV in atrial preparations (P < 0.05). 4. Inhibition of the Na+ pump by DHO increased aNai of the atrium within 10 min by 0.6 +/- 0.1 (n = 7), 1.3 +/- 0.1 (n = 5) and 3.2 +/- 0.2 mM (n = 5) at 5, 10 and 30 microM, respectively. In the papillary muscle, 10 microM DHO was without effect while aNai rose by 1.0 +/- 0.1 (n = 5) and 2.9 +/- 0.2 mM (n = 6) at 30 and 120 microM DHO. 5. Consistent with the aNai measurements, the potency of DHO to increase force of the isometric contraction was three times higher in atrium than in papillary muscle (stimulation frequency 0.2 Hz). 6. Hydrolytic activity of sarcolemmal Na+,K(+)-ATPase isolated from atria amounted to only one third of that detected in ventricles (0.07 +/- 0.01, n = 6, versus 0.2 +/- 0.01 mumol phosphate released min-1 (g tissue)-1, n = 5). The inhibitory potencies of DHO on sarcolemmal Na+,K(+)-ATPase preparations were found to be identical in the enzymes from either tissue. 7. It is concluded that a lower Na+ pump density is responsible for the higher aNai and for the lower resting membrane potential in atrial as compared to ventricular cells. The regulation of cellular Na+ homeostasis in atrial muscle appears to be closer to the limits of its capacity than in ventricle, explaining the higher sensitivity of the atrium to interventions which impede Na+ pump activity. PMID- 8229861 TI - Differences between contractions in vitro of slow and fast rat skeletal muscle persist after random reinnervation. AB - 1. The medial popliteal nerve was sectioned in adult rats, and reinnervation was permitted after nerve reunion. Reinnervation was observed in soleus (slow twitch) and plantaris (fast twitch) muscles for periods of 4-21 months after the initial operation. 2. In the reinnervated muscles, isometric twitch time to peak (contraction time) and time to half-relaxation were longer in soleus than plantaris in every muscle examined. 3. Some of the muscles were stained for actomyosin ATPase activity. The proportion of type I fibres (presumed slow) was at least twice as high in soleus compared with plantaris muscles. The proportion in the latter was similar to that predicted from random reinnervation. 4. The effects of differing proportions of fast and slow fibres on the twitch contraction and half-relaxation times were modelled, and the experimental twitch data were found to be compatible with the proportions of fibre types in individual muscles. 5. It is speculated that the mechanism accounting for the higher proportion of slow fibres in soleus muscle probably does not involve selective reinnervation. It is suggested that fast motoneurones may be transformed to slow by a trophic chemical released by slow muscle fibres; such a process would be more extensive in predominantly slow twitch soleus than a fast muscle, and the resulting decrease in the proportion of fast motoneurones would account for the less extensive conversion in soleus. PMID- 8229862 TI - The electrical geometry, electrical properties and synaptic connections onto rat V motoneurones in vitro. AB - 1. We have developed a tissue slice preparation which allows the study of the actions of single presynaptic neurones onto single trigeminal motoneurones in the immature rat. Our aim in this first stage of the work has been to assess the validity of this preparation as a model for responses obtained in vivo from trigeminal motoneurones in adult rats. We have quantified the integrative properties of the motoneurones and also the variability in transmission at synapses of single presynaptic neurones onto the motoneurones. This data has then been compared to similar published data obtained from adult (rat) trigeminal motoneurones in vivo. 2. Quantitative reconstructions were made of the morphology of three motoneurones which had been labelled with biocytin by intracellular injection. The neurones gave off six to nine dendrites, of mean length 522 microns (S.D. = 160; n = 22), which branched on average 10.5 times to produce 11.45 end-terminations per dendrite (S.D. = 8.57; n = 22). The mean surface area of the dendrites was 0.92 x 10(4) microns2 (S.D. = 0.67; n = 22), and, for individual cells, the ratio of the combined dendritic surface area to the total neuronal surface area ranged from 98.3 to 99.2% (n = 3). At dendritic branch points the ratio of the summed diameters of the daughter dendrites to the 3/2 power against the parent dendrite to the 3/2 power was 1.09 (S.D. = 0.21; n = 217), allowing branch points to be collapsed into a single cylinder. The equivalent cylinder diameter of the combined dendritic tree remained approximately constant over the proximal 25-40% of the equivalent electrical length of the dendritic tree and then showed tapering. The tapering could be ascribed to termination of dendrites at different electrical distances from the soma. 3. Electrical properties were determined for a total of eighty-seven motoneurones, all with membrane potentials more negative than 60 mV (mean = 66.0 mV; S.D. = 5.2) and spikes which overshot zero (mean spike amplitude = 77 mV; S.D. = 10.5; n = 87). The spikes were followed by after-hyperpolarizations (AHPs) of mean amplitude 2.2 mV (S.D. = 1.7; n = 47), and mean duration 54.1 ms (S.D. = 9.5; n = 47). The mean input resistance of the neurones was 7.5 M omega (S.D. = 2.5; n = 69), the mean membrane time constant was 3.5 ms (S.D. = 2.2; n = 35), and the mean rheobase was 1.6 nA (S.D. = 1.1; n = 56).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229863 TI - Effect of medium tonicity on transepithelial H(+)-HCO3-fluxes in rat proximal tubule. AB - 1. The effect of luminal and capillary perfusion with hypotonic or hypertonic solutions containing 25 mM NaHCO3 or NaH2PO4 plus NaCl, K+, Ca2+, Mg2+ and acetate at an osmolality of 100 or 500 mosmol kg-1 on rat proximal H+ secretion was estimated by monitoring luminal pH with Sb microelectrodes. The results were compared to perfusions with the same ionic concentration in which tonicity was adjusted to 300 mosmol kg-1 with raffinose. 2. The kinetics of acidification of luminally injected bicarbonate buffer permits evaluations of H(+)-HCO3-fluxes as well as stationary pH gradients; the kinetics of alkalinization of luminally injected acid phosphate buffer indicates H(+)-HCO3-backfluxes from blood to lumen. 3. In alkalinization experiments, luminal perfusion with hypotonic solution during presence of blood in capillaries or hypotonic capillary perfusion leads to a decrease of stationary pH, an increase of alkalinization half-time and consequently a decrease of passive H(+)-HCO3-backflux. 4. In alkalinization experiments, during luminal and/or capillary perfusions with hypertonic solutions, no significant differences in the stationary pH, alkalinization half time and H(+)-HCO3-backflux were found. 5. During acidification experiments, with both hypo- and hypertonic perfusions, no significant differences in stationary pH, acidification half-time and H(+)-HCO3-flux were observed. 6. Luminal perfusion with hypotonic solution increases specific epithelial resistance in the presence of blood in capillaries. Luminal perfusion with hypertonic solution does not change this parameter. 7. Volume changes, measured by the split-drop method, are slow during the first 30 s and do not explain the increased alkalinization half-time during luminal perfusion with hypotonic solution, since this is the period of fastest pH change. 8. Luminal perfusion with hypotonic solution decreases apparent H+ permeability in the presence of blood or hypotonic solution in capillaries. Hypertonic solutions in all experimental conditions had no significant effect on this parameter. 9. The data indicate that decrease of tonicity of fluids in contact with proximal tubule epithelium affects passive H(+)-HCO3-backflux, which proceeds in part through the shunt path, while acidification (H+ secretion), which is transcellular, is not affected by extracellular tonicity. PMID- 8229864 TI - Classification and diagnostic criteria for oral lesions in HIV infection. EC Clearinghouse on Oral Problems Related to HIV Infection and WHO Collaborating Centre on Oral Manifestations of the Immunodeficiency Virus. AB - A consensus has been reached on the classification of the oral manifestations of HIV infection and their diagnostic criteria, based on presumptive and definitive criteria. The former relate to the initial clinical appearance of the lesion and the latter are often the result of special investigations. Candidiasis, hairy leukoplakia, specific forms of periodontal disease [linear gingival erythema, necrotising-(ulcerative) gingivitis and necrotising(ulcerative) periodontitis], Kaposi's sarcoma and non-Hodgkin's lymphoma are strongly associated with HIV infection. Lesions less commonly associated with HIV infection and lesions seen in HIV infection, but not indicative of the disease, are also listed. PMID- 8229865 TI - Oral Kaposi's sarcoma: a 10-year retrospective histopathologic study. AB - Microscopic diagnosis of early Kaposi's sarcoma continues to be a challenge to the pathologist, as does the identification of bacillary angiomatosis (BA) which may have a similar appearance. 120 oral Kaposi's sarcoma (KS) biopsies submitted to the UCSF oral pathology service from 1981-1991 were reviewed in order to describe the clinical-pathologic spectrum of these lesions and to search for unrecognized cases of BA. Also, histopathologic features of oral KS were compared to 30 oral pyogenic granulomas, and immunohistochemical stains for endothelium associated CD34 antigen were done. The diagnosis of KS was confirmed in all biopsies and no cases of BA were found. Histologically, the KS specimens exhibited numerous features that separated them from pyogenic granulomas, and could themselves be divided into two clinical-pathologic subtypes: small, well delineated macular lesions (31), which were characterized by inconspicuous patches of spindle cells containing ill-defined vascular spaces; and larger, infiltrative nodular lesions (89), which were characterized by spindle cells lining vascular slits and bizarre-shaped vessels. Extravasated RBCs were evident in almost all KS lesions; hemosiderin deposits and hyaline globules were seen in half of each of the small and large lesions. Nuclear atypia was minimal and mitotic activity was slight. Lymphocytes in small lesions added to the difficulty of microscopic interpretation of these incipient lesions. CD34 was expressed on all spindle cells lining vascular spaces in larger lesions and on spindle cells of small, subtle lesions. We conclude that within the spectrum of lesions that are diagnosed as oral KS, two clinical-pathologic types can be identified: macular small spindle-cell lesions and nodular infiltrative vascular lesions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229866 TI - Keratinocyte expression of calprotectin in oral inflammatory mucosal diseases. AB - Oral mucosal inflammation evolves in response to microbial pathogens and non infectious antigens which activate humoral and cell-mediated immunologic processes. Most of these disease processes invoke a leukocyte response culminating in cellular infiltration of the submucosa and, to some degree, transmigration into the epithelium itself. Calprotectin, a leukocyte-derived dimeric protein complex that has potent antibacterial and antifungal effects, has recently been identified in skin and mucosal keratinocytes implying that epithelium may biochemically contribute to the overall mechanism of host defense. In this study, the upregulation of calprotectin as assessed immunohistochemically is pursued for oral diseases of immunopathologic, fungal and viral origin. In lichen planus, candidiasis, herpes virus stomatitis, and oral hairy leukoplakia, calprotectin was found to be expressed to a significantly higher level than in normal control mucosal samples. PMID- 8229867 TI - Orofacial manifestations in primary immunodeficiencies: T lymphocyte defects. AB - The orofacial manifestations of 10 non-HIV-infected children with primary defects of cell mediated immunity are reported. Oral candidiasis was observed in 80% of patients, with oral ulceration present in 50%. Other oral manifestations included enamel hypoplasia, angular stomatitis, oral purpura and recurrent herpetic infection. PMID- 8229868 TI - Orofacial manifestations in primary immunodeficiencies: polymorphonuclear leukocyte defects. AB - The orofacial manifestations of 17 children with primary defects of polymorphonuclear leukocyte function are reported. Oral disease was commonly found and included recurrent oral ulceration (35% of patient group), oral candidiasis (29%), recurrent herpes labialis (12%), enamel hypoplasia (18%) and delayed tooth eruption (12%). Only 3 children had no orofacial disease. Cervical lymph node enlargement was present in 41% of the children. PMID- 8229869 TI - Differentiation of sugar assimilation characteristics and colony phenotypes in pathogenic and commensal oral candidal isolates. AB - A comparison of sugar assimilation patterns has been made using 72 oral candidal isolates recovered from diseased and non-diseased patients. Significant differences were demonstrated between isolates recovered from diseased versus healthy mouths. In addition, significant changes in colony phenotype (switch frequency and morphologic predominance) were found to relate to predictive data generated from carbon source utilization data. These results suggest that biochemical properties and colony phenotypic characteristics may hold promise in predicting the behavior of oral candidal isolates in disease. PMID- 8229870 TI - Low-grade adenocarcinoma of the oral cavity: polymorphous or papillary? AB - Polymorphous low-grade adenocarcinomas are low-grade malignant salivary-gland neoplasms with a broad variety of architectural patterns. Whether papillary structures fall within this spectrum or whether they should lead to a diagnosis of papillary low-grade adenocarcinoma is a subject of debate. This problem was investigated by analyzing a series of 22 salivary gland neoplasms. It was concluded that papillary structures form part of the histologic spectrum of polymorphous low-grade adenocarcinoma but that there are also tumors with papillary structures as a single component. In most instances these are more properly classified as papillary cystadenocarcinomas. As the designation papillary low-grade adenocarcinoma has been used for polymorphous low-grade adenocarcinomas with papillary structures as well as for papillary cystadenocarcinoma, it is advocated that the diagnosis of papillary low-grade adenocarcinoma should be discontinued. PMID- 8229871 TI - Periosteal Gaucher-like cells in beta-thalassemia major. AB - A 24-year-old Chinese woman who has undergone staged surgery for craniofacial deformity secondary to beta-thalassemia major is presented. Local clusters of Gaucher-like cells were found in the periosteum of the mandible. The histologic and ultrastructural features of these cells are described and the pathogenesis and differential diagnoses discussed. To the best of our knowledge, this is the first reported case of Gaucher-like cells occurring outside the lymphohematopoietic system in thalassemic patients. PMID- 8229872 TI - Soft tissue chondroma of the cheek. AB - Soft tissue chondromas are rare benign tumours unrelated to bone that arise primarily in the distal extremities. Lesions in the soft tissues of the oral cavity are extremely rare although several chondromatous lesions have been reported in the tongue. A case is presented of a chondroma arising in the buccal mucosa. It was composed of a lobulated mass of myxoid tissue showing central areas of chondroid differentiation. Immunocytochemistry confirmed the lesion to be mesenchymal and not epithelial in origin. In the differential diagnosis it is important to exclude chondromatous change in a pleomorphic adenoma and also the possibility of a metastasis from an osseous chondrosarcoma. PMID- 8229873 TI - [History in development of electron microscopies at the Department of Physiology, Kyoto University Medical School--contribution of Dr. Sasakawa and Dr, Azuma]. PMID- 8229874 TI - Adaptation to removable prosthesis: a clinical report. PMID- 8229875 TI - Removable partial denture fabrication using extracoronal resilient attachments: a clinical report. PMID- 8229876 TI - Combined percutaneous-permucosal titanium implants for retention of a maxillary prosthesis: a clinical report. PMID- 8229877 TI - A multidie technique for the fabrication of porcelain laminate veneers. AB - Porcelain veneers are a challenge for the dentist and dental technician when they are an integral part of complex restorative treatment plans. A multidie technique is described that allows construction of laminate veneers on removable refractory dies in a stone master cast. This system provides flexibility and accuracy with less chair time when used in combination with epoxy resin dies. Multiple restorative techniques can be used simultaneously with one master cast, and emphasis is directed to harmonious working relationships between the dentist, dental technician, and patient. PMID- 8229878 TI - Effect of an adhesive primer on the integrity of occlusal veneer-metal interface and wear of composite resin veneered restorations. AB - This study examines the influence of a metal-resin bonding agent on the wear rate of composite resin veneer. The specimens were prepared with a silver-based casting alloy and a light-cured composite veneering material. After application of repeated loading through a three-body wear testing device, the amount of wear was measured. The results showed no significant difference in wear between bonded and nonbonded specimens on the 1.5 mm areas of the veneer-metal interface. However, the bonded specimens exhibited significantly lower wear values at the veneer-metal interface. The use of an adhesive primer effectively enhanced the integrity of the occlusal veneer-metal junction. PMID- 8229879 TI - Clinical factors affecting adhesive fixed partial dentures: a 6-year study. AB - One hundred forty-one adhesive fixed partial dentures were placed under controlled conditions for a 6-year longitudinal study. The influence of location (anterior/posterior, maxilla/mandible), tooth preparation techniques (retentive/less invasive), and four different methods of metal conditioning (sandblasting/electrolytic etching and/or pyrolytic/tribochemical silane coating) on the survival rate was investigated. Failures (23 of 24) were caused by loss of adhesion at the metal-cement interface. The retentive tooth preparation reduced the risk of failure to almost one twentieth, but the intraoral location did not influence survival time. The effect of silane-coating on longevity was extremely positive and was not reflected by successful retainers. PMID- 8229880 TI - Evaluation of two dentin adhesives in cervical lesions. AB - The clinical effectiveness of two dentin adhesives, Clearfil New Bond and Scotchbond 2, was evaluated in two different cavity designs. Group A was without enamel bevel or acid etch and with a butt-joint cavity; and group B had enamel bevel, acid etch, and feather-edged cavities. The retentive rate and marginal adaptation were monitored for 2 years. In the Clearfil system, 21% of group A restorations failed after 2 years, whereas virtually all the group B restorations (99%) were retained. In addition, after 2 years, the total of debonded group A restorations in the Scotchbond 2 system expanded to 13%, whereas no restorations from group B were lost. There was clearly marginal deterioration in time irrespective of the bonding system. Nevertheless, the marginal adaptation of cervical lesions restored with Clearfil New Bond adhesive in combination with Clearfil Ray composite resin revealed fewer defects compared with the Scotchbond 2 adhesive with Silux Plus composite resin restorations. SEM evaluation disclosed composite resin remnants on the dentin surface in cavities with lost fillings, which indicates partial cohesive failure of the adhesive joint. PMID- 8229881 TI - Definitive diagnostic waxing with light-cured composite resin. AB - The benefits of perceptive treatment planning for prosthodontics are discussed, and a method of developing diagnostic waxing with light-cured composite resin is described. The advantages of this programmed approach are illustrated with clinical examples. PMID- 8229882 TI - Reliability of the evaluation of occlusal contacts in the intercuspal position. AB - Reliability of clinical measurement is essential to any clinical discipline. This investigation assessed intraexaminer and interexaminer reliability achieved in identifying contacting teeth in the intercuspal position. Shim stock and an articulating film were compared in the evaluation of occlusal contacts of 337 antagonist occlusal pairs in 24 young adults by two examiners. Results were compared by use of a simple proportion of agreement and, when possible, the Kappa statistical test that corrects for chance agreement. Shim stock displayed better reliability than articulating film and appeared suitable for clinical measurement of occlusal contacts in intercuspal position. PMID- 8229883 TI - Prosthodontic treatment activities in the Canadian Forces Dental Services. AB - Prosthetic treatment activities and dental laboratory support are profiled to characterize the influence that a structured, publicly funded military program has on the delivery and practice of prosthetic dentistry. Changing oral health trends and advances in techniques and materials have had quantitative and qualitative impacts on the practice of prosthodontics in the Canadian Forces Dental Services over the past 25 years. PMID- 8229884 TI - Retention designs for bone-anchored facial prostheses. AB - A facial prosthesis can be retained by bone-anchored implants and by a retentive bar. Because tissue response is critical around the abutments, the retentive bar should be constructed so that it is comfortable, conveniently hygienic, and designed without compromising the contours of the prosthesis. PMID- 8229885 TI - Treatment of lagophthalmos of the eye with a custom prosthesis. AB - Inability to close the eyelids (lagophthalmos) is a major problem for patients with facial paralysis. Various treatment modalities and combinations can be used to restore function and esthetics. Lid loading is an effective method of treatment and can be further enhanced by custom fabrication of the prosthesis. The dentist functions as an integral part of the treatment team, working with the plastic surgeon and ophthalmologist to achieve the most functional and esthetic result. PMID- 8229886 TI - Investigation of the effects of a variable lateral guidance incline on the pantronic registration of mandibular border movement: Part II. AB - This study was designed to test the hypothesis that changes in lateral guidance inclination could affect the condylar registrations in the recordings of an electronic pantography. In seven healthy subjects, pantronic registrations of immediate side shift, progressive side shift, and orbiting and protrusive condylar paths were recorded with lateral anterior guidance inclinations of 0, 20, 30, 45, and 60 degrees from the same pin height. Analysis of variance with repeated measures showed a significant decrease in the immediate side shift (p 0.016) and the progressive side shift (p 0.051) as the lateral guidance angle increased and no significant effect on the orbiting registrations. These observations, together with those from other studies, suggest that the concept of fabricating the working-side tooth guidance exactly according to instrument condylar settings (which themselves may be subject to a variable recording lateral anterior guidance) appears untenable and demands further study. PMID- 8229887 TI - Prevalence of temporomandibular disorders in relation to some occlusal parameters. AB - A clinical examination survey included 600 women and men between 18 and 22 years of age studied for the possible association between temporomandibular disorders and a number of occlusal parameters (Angle's classification, type of eccentric occlusion, chewing side preference, horizontal overlap value, and non-working side contact occurrence. Extraoral and intraoral examination was performed for each subject, and horizontal overlap value was determined by the use of a dial caliper gauge. The presence of non-working contacts was detected by use of the dental floss technique. The results showed high dependence between the frequency of temporomandibular disorders and class II division 1, class III patients, group function occlusion, and a high horizontal overlap value. Furthermore, class II division 2 patients showed a reduced occurrence of temporomandibular disorders in relation to the other classes. When the non-working side contacts were increased, a higher association of temporomandibular disorders was observed. PMID- 8229888 TI - Radiopacity of posterior composite resins, composite resin luting cements, and glass ionomer lining cements. AB - The radiopacities of seven posterior composite resins, seven composite resin luting cements, and six glass ionomer lining cements were studied. The purpose was to obtain an indication of radiopacity values of different brands within each of these three groups of materials and to show differences in radiopacities of composite resin inlay materials and their accompanying luting cements. On radiographs, the optical densities of standardized samples were determined and radiopacity values of the materials were expressed in millimeter equivalent aluminum. The composite resins observed displayed higher radiopacities than aluminum. A luting cement and a glass ionomer cement were shown to be less radiopaque than aluminum. Generally the radiopacity of composite resins appeared to be higher than their accompanying luting cements. If materials with substantial differences in radiopacity are used in combined applications for the restorative treatment of teeth, lower radiopacity can interfere with the diagnosis of caries and the detection of gaps near the restoration. PMID- 8229889 TI - Noise level of dental handpieces and laboratory engines. AB - Impaired hearing in dental practitioners has been the subject of various investigations. Although individual age and susceptibility can play a significant role, the intensity of the noise, distance from the noise source, and total duration of the noise are the important factors in hearing. In this study the noise levels of different handpieces and laboratory engines were evaluated. Handpiece noise measurements were made while instruments were running free and during operation with various cutting tools. These measurements were performed with two kinds of noise-measuring equipment: (1) a precision sound level meter and (2) a noise recording and analyzing circuit. The results indicated that (1) there were significant differences among the noise levels of the various dental engines used, (2) the low-speed, angled-design handpiece has the lowest noise level, whereas the laboratory engine has the highest, (3) the noise level increases during cutting was compared with noncutting, and (4) there was no significant difference between the two methods of measurement. PMID- 8229890 TI - Recent graduates' and current dental students' evaluation of their prosthodontic curriculum. AB - This survey of recent graduates and third- and fourth-year dental students at Indiana University School of Dentistry determined whether the prosthodontic curriculum adequately prepared students for the prosthodontic portion of dental practice and whether there was material that should be added to or omitted from the prosthodontic curriculum. The survey evaluated 106 topics in the prosthodontic curriculum and areas related to prosthodontics. On the basis of their importance to the respondents, 59.4% of the topics were considered adequately emphasized, 2.9% of the topics were overemphasized and should be reduced in emphasis or eliminated, and 37.7% of the topics were underemphasized, which indicated the need to increase time devoted to these topics. Most of the underemphasized topics dealt with new materials, alternate techniques, private practice and its management, and the prosthodontic needs of special types of patients. PMID- 8229891 TI - An alternate light-cured transfer coping material. AB - Transfer copings, used in many aspects of fixed and removable prosthodontics, are commonly made with autopolymerizing resins. Fabricating accurate and dimensionally stable transfer copings with these materials can be difficult. Luma Rez resin, a visible light-cured resin, helps overcome these fabrication difficulties. A technique that uses this material to make transfer copings is described. PMID- 8229892 TI - Integrating fixed and removable provisional restorations. AB - When both fixed and removable components are required, provisional restorations can be difficult. This article describes a technique where provisional restoration "shells" are made with a plastic stent of the diagnostic wax-up on lightly prepared teeth of a diagnostic cast. An acrylic removable partial denture is then waxed to the shells. The shells are relined intraorally after tooth preparation with the partial denture in place. PMID- 8229893 TI - A simplified method to record centric relation when making occlusal splints for dentulous patients. PMID- 8229894 TI - Commissural neurons of the electrosensory lateral line lobe of Apteronotus leptorhynchus: morphological and physiological characteristics. AB - Extracellular injections of horseradish peroxidase were used to label commissural cells connecting the electrosensory lateral line lobes of the weakly electric fish Apteronotus leptorhynchus. Multiple commissural pathways exist; a caudal commissure is made up of ovoid cell axons, and polymorphic cells' axons project via a rostral commissure. Intracellular recording and labeling showed that ovoid cells discharge spontaneously at high rates, fire at preferred phases to the electric organ discharge, and respond to increased receptor afferent input with short latency partially adapting excitation. Ovoid cell axons ramify extensively in the rostro-caudal direction but are otherwise restricted to a single ELL subdivision. Polymorphic cells are also spontaneously active, but their firing is unrelated to the electric organ discharge waveform. They respond to increased receptor afferent activity with reduced firing frequency and response latency is long. Electrical stimulation of the commissural axons alters the behavior of pyramidal cells in the contralateral ELL. Basilar pyramidal cells are hyperpolarized and nonbasilar pyramidal cells are depolarized by this type of stimulation. The physiological results indicate that the ovoid cells participate in common mode rejection mechanisms and also suggest that the ELLs may function in a differential mode in which spatially restricted electrosensory stimuli can evoke heightened responses. PMID- 8229895 TI - Conductances contributing to the action potential of Sternopygus electrocytes. AB - In Sternopygus macrurus, electrocyte action potential duration determines the electric organ discharge pulse duration. Since the electric organ discharge is a sexually-dimorphic behavior under the control of steroid hormones, and because electrocyte action potential durations can range from 3-14 ms, the electrocytes provide a unique opportunity to study how sex steroids regulate membrane excitability. In this study, the voltage-sensitive ionic currents of electrocytes were identified under current- and voltage-clamp as a prelude to further studies on their regulation by sex steroid hormones. Bath application of TTX completely abolished the spike and eliminated an inward current under voltage clamp, indicating that the action potential is due primarily to a sodium current. Calcium-free saline had no effect on spike waveform or voltage-clamp currents, indicating that neither calcium nor calcium-dependent currents contribute to the action potential. Application of potassium channel blocking agents, such as tetraethylammonium and cesium ions, caused changes in the spike which, together with voltage-clamp results, indicate the presence of two potassium currents: an inward rectifier and a classical delayed rectifier. In addition, these cells have a large, presumably voltage-insensitive, chloride current. Differences in one or more of these currents could be responsible for the range of action potential durations found in these cells and for the steroid-mediated changes in spike duration. PMID- 8229896 TI - Perception of complex sounds by the green treefrog, Hyla cinerea: envelope and fine-structure cues. AB - 1. The envelope periodicity of communication signals is an important feature distinguishing advertisement and aggressive calls for the green treefrog (Hyla cinerea). Envelope periodicity, a cue for periodicity pitch perception in humans, is affected by the fine-structure of the signal, a cue for timbre perception in humans. The present study examined perception of two acoustic features affecting waveform fine-structure--harmonic structure and phase structure--in male green treefrogs. 2. We analyzed evoked vocal responses of male green treefrogs living in laboratory arenas to playbacks of digitally-generated signals resembling either conspecific advertisement or aggressive calls in their first harmonic periodicity. Systematic changes in the harmonic structure of these signals were achieved by varying the harmonic relations between frequency components in the signals, and changes in phase structure were achieved by varying the starting phases of harmonically-related components. 3. Calling was significantly influenced by the first harmonic periodicity of the signals. Males vocalized more to signals with the periodicity of the advertisement than the aggressive call. There were no differences in response to harmonic and inharmonic signals with similar spectral content. Phase structure did not significantly influence vocal responses. 4. These results suggest that the fine-structure ("timbre") of complex acoustic signals is not a significant feature guiding behavior tested using a communication response in this species. PMID- 8229897 TI - Acoustic intensity discrimination by the cichlid fish Astronotus ocellatus (Cuvier). AB - The acoustic intensity discrimination ability of the oscar (Astronotus ocellatus), a cichlid fish, was investigated using an automated positive reward method. Intensity discrimination thresholds (delta I, in dB) for 7-s continuous pure tone signals were measured both as functions of sound intensity above thresholds, i.e., sensation levels, (SL)(+10 dB, +20 dB and +30 dB) and frequency (200 Hz, 500 Hz, and 800 Hz). delta I at 500 Hz for +10 dB, +20 dB, and +30 dB SLs are 8.9, 5.5, and 3.3 dB, respectively. delta I (at +20 dB SL) for 200 Hz, 500 Hz, and 800 Hz are 4.5, 5.5, and 9.3 dB, respectively. Despite having poor auditory sensitivity (narrow frequency range and high thresholds), the intensity discrimination ability of the oscar follows the general trends of previously studied fish species, however, with higher thresholds. PMID- 8229898 TI - A graph theory model of the semantic structure of attitudes. AB - The semantic structure underlying the attitudes of pretreatment and posttreatment drug addicts was modeled using a network analysis of free word associations. Measures of graph theoretic properties were used to assess structural differences in the associative networks of the two populations. These measures modeled the information processes of associative networks proposed in the spreading activation theory of semantic processing. As expected based on graph theory, the structure of the associative networks of posttreatment subjects was more dense, less constrained, and more hierarchically organized by the self concept. In a test of the network model, the subjects' evaluations of concepts in the associative network were found to be a function of their evaluations of semantically similar concepts. Although preliminary and limited, the results suggest that graph theory may provide a broad mathematical foundation for diverse models of cognitive systems. PMID- 8229899 TI - Computerized assessment of verbal skill. AB - This paper describes a computerized diagnostic test for the assessment of basic literacy skills in Dutch. Central in this test is a skill labeled word image, referring to the subject's implicit knowledge of orthographic structure. Test norms were obtained in a group of reference subjects. The most notable trend found in the standardization study was that the development of word image performance showed a nonlinear, U-shaped trend. Two case studies are reported. A supplementary validation study, based on test data collected from standard reference groups of normal and poor readers, demonstrated that word image performance is significantly related to between-group differences. PMID- 8229900 TI - The psychosocial impact of HIV infection in women. AB - The aim of the investigation was to study the prevalence of current and past psychiatric morbidity and psychosocial problems in HIV seropositive (HIV +ve) asymptomatic women. A cross-sectional controlled study including 57 HIV +ve women belonging to CDC group II and III (43 intravenous drug users and 14 non-IVDUs heterosexuals) and 23 HIV -ve women (15 intravenous drug users and 8 non-IVDUs heterosexuals) is reported. Outcome measures included, past psychiatric history, current psychological status (Zung Anxiety and Depression scales, Symptom Check List 90-Revised), Social Supports and Locus of Control Scales, and information on changes in work, social and sexual life after HIV testing. Results showed that HIV +ve women differed very little from HIV -ve controls regarding outcome measures and indeed for some variables HIV infected women had lower levels of psychological morbidity. Multiple regression analyses showed that alcohol misuse and a predominantly external locus of control accounted for the 29% of the variance of psychiatric distress (F = 9.23, p < 0.0006). The implications of the findings are discussed. PMID- 8229901 TI - Childhood asthma: mothers' affective attitude, mother-child interaction and children's compliance with medical requirements. AB - Maternal critical attitude (CA) and negative mother-child verbal interaction in families with an asthmatic child and in controls were investigated in the present study. Twenty-eight children with bronchial asthma (6-13 yr) and 23 healthy children with their mothers participated in the project. Mothers were asked to describe their child (Five Minute Speech Sample; FMSS). Mother and child were then asked to discuss a mutual problem (Problem Discussion; PD). Tape recordings were taken. The mothers' responses during the FMSS were used to assess CA. Negative verbal behavior during PD was coded by means of a reliable coding system for dyadic interaction. Significantly more mothers of the asthmatic children than of the controls showed CA during the FMSS. Mothers of the asthmatic children made significantly more critical remarks and offered more negative solutions during the PD. Within the asthma group mothers who made more critical remarks had to make more effort to have their child comply with the medical requirements. Additionally, mothers showing a higher total amount of negative verbal behavior had children with rather high IgE levels. There was no systematic difference between children having CA mothers with high vs low face-to-face contact. PMID- 8229902 TI - Gender differences in the psychosocial variance of Framingham and Bortner Type A measures. AB - The object of the present study was to determine whether the non-genetic variance of a Type A scale composed of Framingham and Bortner items had gender specific psychosocial components. The study was performed on a group of Swedish twins so that variance explained by heritability for Type A could first be removed from the equation. The overall Type A score had been found to relate to self-reported CHD in this population. The dependent variable was the standardized score residual remaining after removing the genetic variance (i.e., that explained by co-twin score and zygosity). Multiple regression analyses revealed that there were differences in the psychosocial components of the Type A residual in men and women. These results are discussed in terms of culturally accepted gender roles and their possible implications for health endpoints. PMID- 8229903 TI - Is raised blood glucose associated with the relief of tension? AB - A series of three studies examined the influence of glucose drinks and increasing blood glucose levels on mood. Experiment one considered the short-term effect of a glucose containing drink on mood in young adults: a glucose drink in the morning and higher blood glucose levels were both associated with feeling less tense. Higher blood glucose was also correlated with greater self-reported energy. In the second experiment high blood glucose levels were sustained for 2 hr by repeatedly taking a glucose containing drink. Again higher blood glucose levels were associated with reports of less tension. In a final study subjects were placed in a frustrating situation. The consumption of a glucose drink resulted in fewer negative responses, following a negative statement from the experimenter, in subjects who had fasted over-night. It was concluded that higher blood glucose levels are associated with feeling less tense. PMID- 8229904 TI - The urethral syndrome: a controlled evaluation. AB - It has long been considered that psychological factors play an important role in women with urinary tract symptoms. Where no significant infection can be demonstrated, psychological factors have been claimed as paramount to the extent that the 'irritable urethral syndrome' has been postulated as a parallel to the irritable bowel. This paper reports on a controlled study of psychological and menstrual problems in women presenting in family practice with urinary symptoms. Women were assessed by structured interview. Two groups of women took part: (A) 54 women with urinary symptoms, 15 of whom later proved to have significant bacterial infection on culture. (B) 21 women attending for contraceptive advice also agreed to participate in a comparison group. Women presenting with urinary tract symptoms had higher mean scores on psychological and menstrual measures than comparison women, regardless of the presence of demonstrable infection. Thus, psychological and menstrual symptoms appear to determine whether a woman who experiences urinary tract symptoms attends her doctor, regardless of whether significant infection is present. No evidence was found, for the existence of an irritable urethral syndrome, in which psychological factors are postulated as the predominant cause. PMID- 8229905 TI - Hypochondriacal attitudes, pain sensitivity, and attentional bias. AB - The relation between hypochondriacal attitudes, thermal pain threshold, and attentional bias toward pain was examined in a non-clinical population (N = 28). Attentional bias was operationalized with a concentration-performance test, which subjects performed while connected to a pain stimulator. Subjects were informed that they would receive a painful stimulus during the second part of the test, while the first part was introduced as pain-free. The pain stimulus was never applied during the test phase. The expectancy of a forthcoming pain stimulus reduced the performance of high hypochondriacal subjects in both parts of the test. Low hypochondriacal subjects, on the other hand, displayed significantly better performance in the first, pain-free compared to the second, pain-related part of the test. Thermal pain thresholds were assessed at four measuring sites (thenar, neck, collar-bone, abdomen), but no relations with hypochondriasis sum scores and locus of pain stimulation were found. A stepwise multiple regression of pain threshold by individual Illness Attitude Scales (IAS) led to 66% of the variance being explained by the scales 'concern about pain', 'worry about illness', and 'disease phobia'. Results are discussed in terms of amplifying somatic style, preoccupation with or attentional bias toward bodily symptoms, and experimental induction of a hypochondriacal state. PMID- 8229906 TI - The measurement of fatigue: a new instrument. AB - Fatigue is a frequent medical symptom which has not been routinely measured. We present a 29-item fatigue assessment instrument, describe its psychometric properties, and use it to differentiate normal fatigue from fatigue related medical disorders. Differences in fatigue across a variety of medical disorders, the reproducibility of the fatigue instrument, and its convergent validity with other fatigue measures are also described. PMID- 8229907 TI - Effects of acute stress on lymphocyte beta 2-adrenoceptors in white males. AB - The effects of acute cold pressor stress on peripheral blood cells and lymphocyte beta 2-adrenoceptor density and affinity were examined in 18 young, healthy normal white male volunteers. Blood samples were drawn two times before and three times after the stressor. The percents of lymphocytes and neutrophils, plasma cortisol and the beta 2-adrenoceptor density were significantly different among blood samples. There were no differences between samples in total white blood cell count, and percent of monocyte, basophil and eosinophil or beta 2 adrenoceptor affinity. The results suggest that cold pressor stress causes lymphocytosis and increases the lymphocytic beta 2-adrenoceptors concentration. Methodological issues, acute vs chronic condition effects, immune cell subset variations and the psychosomatic activity-affect model were discussed. Such adrenoceptor changes may provide one mechanism by which stress can affect the onset of cardiac/hypertensive disorders. PMID- 8229908 TI - The effects of sodium loading on blood pressure and pain responses to the cold pressor test. AB - Two cold pressor tests were administered to 18 healthy normotensive males on two occasions, after 2 weeks of dietary sodium loading and after 2 weeks of maintaining their normal diet. While the addition of an extra 10 g/day of sodium to the diet had no overall effect on resting cardiovascular activity, it produced significantly greater diastolic blood pressure and smaller heart rate responses to the pain stimuli. As well, consistent with the large animal literature indicating a relationship between high blood pressure and diminished pain sensitivity, pain ratings were found to be significantly lower during the sodium loaded testing session. The ratings were corroborated by evidence of lower skin conductance reactivity to the second cold pressor test in the sodium loaded condition. Information concerning an elevation of risk for hypertension, perhaps provided by baroreceptors, may lead to compensatory reactions with hypoalgesia as one effect. PMID- 8229909 TI - When a colleague commits suicide. Interview by Bruce P. Mericle. AB - 1. When a fellow nurse commits suicide, we experience a range of feelings not unlike others (including our patients) feel. 2. As psychiatric nurses, we may deny our feelings in order to appear normal and/or in control. Because of our professional roles we may want to be viewed as being immune to mental problems. 3. When a colleague dies, it is important that we heed the advice we often give our patients. Talk about your experience and allow yourself to feel whatever it is you are feeling. We will probably feel guilty, but we must realize that the person who commits suicide is ultimately responsible for that act. 4. Psychiatric nursing should promote efforts that encourage professionals to seek mental health services when they are needed. If it is okay to have problems, then it is okay to seek help. PMID- 8229910 TI - Road to recovery. Post-traumatic stress disorder: the hidden victim. AB - 1. Post-traumatic stress disorder (PTSD) is characterized by the development of a set of specific symptoms following a psychologically distressing event that is outside the range of usual human experience. 2. Recovery does not seem to feel as difficult or overwhelming if it is approached "one day at a time." 3. The psychologic damage that occurs in PTSD is due primarily to holding the experiences inside, "stuffing" them down through various psychodynamics, or keeping the experiences subdued through addictive behaviors. 4. PTSD among Vietnam veterans is not a mental illness; it is a reaction to extreme stress and a reaction to keeping memories stuffed down and not allowed to surface. PMID- 8229911 TI - Use of art therapy with post-traumatic stress disordered veteran clients. AB - 1. The post-traumatic stress disordered veteran client may exhibit a cluster of problematic behaviors that are integral to current maladaptive coping patterns. 2. These coping patterns can be successfully addressed through a multidisciplinary therapeutic approach that emphasizes client-controlled expression of emotions and client-focused step-by-step behavior modification. 3. Art therapy and nursing, in a collaborative approach, can facilitate individual and group interventions that promote expression of feelings, congruency between experience and self-concept, and feelings of effectiveness in behavioral change. PMID- 8229912 TI - A retrospective study of patient falls in a psychiatric hospital. AB - 1. While falls on medical-surgical units are the focus of extensive research, falls on inpatient psychiatric units are an understudied critical event. 2. The purposes of this study were to identify the variables associated with psychiatric patient falls and to use that information to assess risk and, therefore, prevent falls in this population. 3. The psychiatric patient at risk for falling is described as a woman with a prior history of falls; less than 65 years of age; experiencing anxiety and agitation; and receiving a sedative, a tranquilizer, and a laxative. Additionally, this patient is more likely to fall in a community area. PMID- 8229913 TI - Codependency: a clear perspective. PMID- 8229914 TI - The North Staffordshire Trauma System. AB - The first trauma centre in the UK has been established at the North Staffordshire Hospital in Stoke-on-Trent. The early development of this centre and the wider system within which it functions is described. The Trauma Centre and corresponding evaluation project by the University of Sheffield have been funded by the Department of Health. The outcome for injured patients treated in the new trauma system over a 4-year period will be compared with outcomes from existing systems in and around comparator sites in Preston and Hull. These findings will inform the debate on the future of trauma care in the UK. PMID- 8229915 TI - Problems associated with ultrasonography of the canine ovary around the time of ovulation. AB - Real-time B-mode ultrasound imaging was carried out, using a scanner of moderate quality, on 40 fresh bitch cadavers to identify the presence of ovaries and significant ovarian structures. The results indicated that accurate identification of the ovaries depended on the presence of significant follicles or corpora lutea (CL). The findings were verified at post-mortem examination. The study was continued by scanning 35 live bitches using three different scanning units of increasingly improved quality to show that the process of ovulation could be assessed with reasonable accuracy with improved quality of equipment, although the exact fate of the follicle could not be imaged. The results were judged against assays for blood circulating progesterone. A final group of seven live bitches were scanned with a high-grade scanner using an annular phased-array transducer to attempt to image the process of ovulation. The non-echogenic follicle disappeared and was replaced by the CL that was hypoechoic with respect to surrounding tissue, thus allowing accurate assessment of the time of ovulation. This was confirmed by progesterone assay. PMID- 8229916 TI - Ultrasonographic appearance of the ovary and uterus of the bitch during oestrus, ovulation and early pregnancy. AB - The ovaries and uteri of ten bitches were imaged daily from the onset of pro oestrus until 30 days after the plasma surge of luteinizing hormone. During pro oestrus small follicles increased in size but did not protrude above the surface of the ovary. Ovulation was characterized by a decrease in the number of fluid filled follicles and their replacement with similarly sized hypoechoic structures. These structures declined in number after ovulation and were replaced by fluid-filled corpora lutea. Gradual thickening of the wall of the corpus luteum occurred with subsequent obliteration of the central anechoic cavity. During pro-oestrus and oestrus there was an increase in the transverse diameter of the uterus, which developed a characteristic appearance with central radiating areas of hyperechogenicity and zones of hypoechogenicity. These uterine changes were not considered to be specific enough to be used as an indication of ovulation. Uterine diameter and echogenicity did not differ between pregnant and non-pregnant bitches until gestational sacs were identified, a mean of 18.6 +/- 0.9 (SD) days after the plasma surge of luteinizing hormone. Embryonic tissue and heart beats were first detected between 23 and 27 days after the pre-ovulatory luteinizing hormone surge. PMID- 8229917 TI - Measurements of faecal oestradiol and progesterone in non-pregnant and pregnant domestic and exotic cats. AB - Faecal samples collected for variable periods from 12 animals and five species of cats were assayed for progesterone and oestradiol content by application of standard radioimmunoassays to aliquots (50 microliters) of methanol extracts (4 ml) of a mixture of 0.5 g sample, 0.5 ml water and 1 g aluminium oxide, following partitioning of the total extract with petroleum ether (3 ml), further dilution of assay aliquots and drying. Recoveries averaged 100 and 72% for oestradiol and progesterone, respectively. Results included increases in progesterone during luteal phases or pregnancies to 7688 ng g-1 (tiger, Panthera tigris), 2594 ng g-1 (lion, P. leo), 3000 ng g-1 (cheetah, Acinonyx jubatus) and 4915 ng g-1 (caracal, Felis caracal). Faecal oestradiol peaks near oestrus included 246 ng g-1 (tiger), 175 ng g-1 (lion) 190 ng g-1 (cheetah), 23 ng g-1 (caracal) and 190 ng g-1 (domestic cat, F. catus). PMID- 8229918 TI - Rapid detection of the preovulatory luteinizing hormone peak in the blue fox (Alopex lagopus) by an enzyme-linked immunosorbent assay. AB - An enzyme-linked immunosorbent assay (ELISA) developed for the rapid measurement of luteinizing hormone (LH) in blood of many mammalian species has been validated in the fox. For this purpose, daily plasma samples collected from six vixens during the periovulatory period were assayed for LH using ELISA. The results were compared with those obtained by radioimmunoassay (RIA), and related to parameters used to determine the time of ovulation (onset of oestrus, vaginal electrical resistance peak) and to ovarian findings. This ELISA provided a highly sensitive measure of LH (125 pg ml-1) from a 10 microliters plasma sample within 3 h. Plasma concentrations of LH measured by ELISA and RIA were significantly correlated (r = 0.911). In the blue fox, ovulation occurs once a year and coincides with the start of oestrus. The LH peak preceded the start of oestrus by 1.5 days (1-3) and preceded the vaginal electrical resistance peak by 2.2 days (1 4). The rapid and precise detection of the preovulatory LH peak by this assay provides a convenient method to determine the time of ovulation and therefore the optimum time for artificial insemination or oocyte collection for fertilization in vitro. PMID- 8229919 TI - Progesterone and oestradiol in canine plasma monitored by enhanced luminescence immunoassays. AB - The Amerlite immunoassay system was evaluated for direct measurement of progesterone and oestradiol in canine plasma. The progesterone assay was evaluated without modification. To increase the sensitivity of the oestradiol assay, the horseradish peroxidase-labelled tracer was diluted 1:2 and the antiserum 1:10. Subsequently, the standards supplied in the kit were altered to produce a new standard curve. The dilution curves of canine plasma samples with high progesterone and oestradiol contents were parallel with the standard curves based on human serum. The relation between measurements of both hormones in canine plasma using established extraction methods as references and the Amerlite procedures were highly comparable, resulting in the linear regression equations y = 0.91x + 1.3 (progesterone, n = 100) and y = 0.79x + 0.9 (oestradiol, n = 84). However, for oestradiol the closest relation to the reference method was achieved when the incubation period was extended to 24 h at 4 degrees C. When measuring oestradiol and progesterone in canine sera during the oestrous cycle the hormone patterns were not influenced by the methods applied. PMID- 8229920 TI - Species-specific features of oestrous development and blastogenesis in domestic canine species. AB - The reproductive physiology of taxonomically closely related species is usually very similar. The main difference in the reproduction of the dog and fox is the length of the different phases of the oestrous cycle. Pro-oestrus and oestrus are longest in the dog: oestrus lasts 3-5 days in the blue fox and 1-3 days in the silver fox, compared with about 1 week in the dog. The profiles of sex steroid concentrations in plasma during oestrus and pregnancy are similar and the luteal phase in non-pregnant animals is prolonged, progesterone concentrations reaching a maximum by 15-30 days after the luteinizing hormone (LH) peak in the dog, by 10 20 days in the blue fox and by 5-15 days in the silver fox. The duration from LH surge to ovulation is about the same in the dog and fox, but thereafter the oocytes and early embryos develop faster in foxes than in the dog. The tubal transport time is 4-6 days in the silver fox, embryos entering the uterus at the 4-16-cell stage. In the blue fox the oocytes remain in the oviducts for 8-10 days, developing into the morula stage, whereas in the dog a still longer oviductal phase results in embryos that are at the compact morula or blastocyst stage when entering the uterus. The preimplantation period is about 1 week in the dog and the blue fox, but 9-10 days in the silver fox.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229921 TI - Identification and immunolocalization of proteins synthesized by dog endometrium and membranes. AB - Previous studies on cyclic and pregnant bitches on dioestrus days 3-10 indicated synthesis de novo of at least ten protein complexes, two of which are major protein groups (cP5, M(r) 54,600; cP6, M(r) 23,000). Protein expression differed by day but not by status, suggesting that the embryo does not affect uterine protein synthesis. Ovariectomized bitches treated with various steroid regimens showed induction of cP5 and cP6 only after oestrogen priming followed by progesterone. Recently, N-terminal amino acid microsequencing has identified cP6 as a member of the retinol-binding protein (RBP) family. The present study was designed (a) to identify and characterize proteins synthesized de novo in explant culture from cyclic (dioestrus days 10-16) and early pregnant (dioestrus days 10 26) dogs, (b) to examine distribution of proteins by endometrium preimplantation (PI), and between (BI) and at (I) implantation sites; (c) to characterize proteins synthesized by embryonal membranes and (d) to localize RBP and CUPED (a cat oestrogen-dependent uterine glycoprotein) immunocytochemically in the uterus and embryonal membranes. Proteins synthesized by endometrium were examined by incorporation of [3H]leucine or [35S]methionine on two-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis followed by fluorography or autoradiography. Nine proteins, characterized by electrophoresis and previously found between dioestrus days 3-10, appeared to be expressed through day 16 of cyclic and day 26 of pregnant dioestrus, although many decreased and cP5 was lost after day 13. In BI and I sites, RBP decreased in number of isoelectric variants as gestation progressed, while the major M(r) form (23,000) was reduced to 21,500. There was no change in M(r) and isoelectric variants of RBP from cyclic dogs, suggesting differences in RBP gene regulation. Proteins synthesized by embryonal membranes appeared to be serum proteins. Immunolocalization of RBP confirmed that oestrogen priming followed by progesterone was required for induction. In pregnant bitches, staining was present in all luminal epithelia on dioestrus day 10, but by day 17 only specific epithelium stained. As pregnancy progressed to day 24 of dioestrus, staining was localized to specific epithelial cells of the deep spongy zone and yolk sac. In ovariectomized bitches, CUPED was confirmed to be oestrogen dependent with staining in both luminal and glandular epithelia.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229922 TI - Acute phase proteins in canine pregnancy (Canis familiaris). AB - An acute phase response has been identified during mid-gestation in bitches by determination of the serum C-reactive protein (CRP) concentration. In four pregnant bitches, a large increase in CRP during mid-gestation was followed by a second increase after parturition. In three pregnant bitches, a similar increase was observed during mid-gestation but the second (post parturition) increase was not detected. In two pregnant bitches only a small rise was observed in mid gestation. No increase in CRP was observed in the serum of two bitches that were not mated following oestrus or in the serum of two bitches that were mated but did not become pregnant. One bitch was diagnosed as pregnant by palpation but did not produce a litter and did not have a mid-gestation increase in CRP. However, this animal did show an acute phase response shortly after oestrus. PMID- 8229923 TI - Regulation of progesterone during pregnancy in the cat: studies on the roles of corpora lutea, placenta and prolactin secretion. AB - The regulation of progesterone secretion and release by the corpus luteum and/or by the placenta in cats during pregnancy has been poorly studied and the relative roles of the corpus luteum and the placenta in the release of progesterone into the plasma is not well defined. It is generally assumed that after 40-45 days of pregnancy, circulating progesterone is mainly produced by the placenta and that after 45-50 days the corpus luteum is no longer necessary to maintain pregnancy, even if remnants of corpora lutea are detected in the ovaries during the following interoestrous period. In this study, the relative roles of the placenta and ovary in progesterone production after 30 days of pregnancy have been analysed in an attempt to define the role of progesterone in pregnant cats. Different experiments involved (1) analysis of the effects of an ovariectomy performed at 30 days of pregnancy, and of megestrol acetate supplementation on plasma progesterone during the second half of pregnancy, (2) analysis of the effects of an ovariectomy performed at 45 days on plasma progesterone production and on pregnancy and (3) analysis of the effects of an antiprolactin agent (cabergoline) at days 30 and 40 of pregnancy on progesterone secretion and pregnancy. These results were all compared with the normal plasma progesterone profiles during (a) pseudopregnancy and (b) pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229924 TI - Echocardiographic study of heart rate in dog and cat fetuses in utero. AB - The heart rate of dog and cat fetuses in utero was studied using real-time B and M mode ultrasonography. Fifteen bitches and 15 queens were monitored from day 19 after the luteinizing hormone (LH) peak in the bitch, and from day 15 after the first mating in the cat, up to parturition, using a 7.5 MHz linear transducer. The first fetal cardiac movements and the cardiac frequency were detected from days 23 and 29 after the LH peak and from days 18 and 25 after mating, in the bitch and in the queen, respectively. In bitches, heart rates were higher [230.2 +/- 15.4 beats per minute (b.p.m.)] than values previously reported. Initial average values were 214 +/- 13.3 b.p.m. and these significantly increased thereafter, with an average of 238.2 +/- 16.1 b.p.m. at day 40. A reduction was noted near parturition. In queens, the average values recorded were 228.2 +/- 35.5 b.p.m. Contrary to what has been observed in the bitch, the cardiac frequencies remained stable during pregnancy. PMID- 8229925 TI - Control of luteal function in the mink (Mustela vison). AB - The ranch mink was studied to determine the role of pituitary luteotrophins on corpus luteum (CL) function before and after implantation. Twelve mink were treated with monoclonal antiserum against gonadotrophin-releasing hormone (GnRH), and 12 with an irrelevant monoclonal antibody during embryonic diapause. Activation of the CL, plasma progesterone concentration and embryo implantation were unaffected by this treatment. In a second trial, groups of ten mink were treated with GnRH antibodies, bromocriptine, bromocriptine plus 0.5 mg prolactin per day per animal, or ethanol vehicle. Comparison of the consequent profiles of progesterone indicated that both bromocriptine and anti-GnRH compromised postimplantation CL function by inducing regression of the corpus luteum. Incubation of dissociated luteal cells from ovaries of mink at 21-24 days after implantation with either LH or prolactin increased the accumulation of progesterone over 2 h. Addition of 25-hydroxy-cholesterol (25OHC) as substrate increased basal levels and the progesterone accumulation stimulated by LH and prolactin; the increases induced by luteotrophins were additive. There was an apparent synergistic interaction between prolactin and canine low-density lipoproteins (LDL) in the stimulation of progesterone secretion in vitro. The results are interpreted to indicate that LH/FSH are not required for luteal support during embryonic diapause, or for luteal activation. Prolactin is necessary for luteal activation, and LH and/or FSH and prolactin are obligate luteotrophins during the postimplantation period in the mink. PMID- 8229926 TI - In vitro fertilization in domestic and non-domestic cats including sequences of early nuclear events, development in vitro, cryopreservation and successful intra and interspecies embryo transfer. AB - The domestic cat may be used as a model for developing assisted reproduction techniques including in vitro fertilization (IVF), embryo culture, cryopreservation and embryo transfer (ET) for application to threatened and endangered species of non-domestic cats. Interoestrous domestic cats were injected with a total of 1.0-6.0 mg follicle-stimulating hormone (FSH) daily for 4 days and with 100 iu human chorionic gonadotrophin (hCG) on day 5. Follicular oocytes recovered at 26 +/- 1 h after hCG were co-incubated for 4-6 h at 38 degrees C in 5% CO2 with spermatozoa (1-2 x 10(5) motile spermatozoa ml-1) collected by artificial vagina. To determine the timing of sperm penetration and early fertilization events in vitro, oocytes were fixed and examined at intervals from 0.5 to 10 h after sperm exposure. The penetration rate of metaphase II (MII) oocytes at 0.5-3 h was equivalent to that at 3-6 h (95 versus 96%). Second polar body extrusion, pronuclear formation and apposition were observed at 2, 6-8 and 10 h, respectively. Simple (Tyrode's) and complex (F-10, M-199 and CMRL-1066) culture media with 10% fetal calf serum were compared for their ability to support development to the morula or blastocyst stage during culture periods of 96-168 h after IVF. The average number of cells per embryo was similar (P > 0.05) in the various media at each interval except that CMRL-1066 reduced (P < 0.05) development at 96 h if it was used before the two-cell stage. In F-10, neither the presence of intact cumulus cells nor changing to fresh F-10 medium at 48 h affected development at 96 h. Blastocyst development at 168 h was similar in both F-10 (18%) and Tyrode's (26%). To determine developmental ability in vivo, IVF derived embryos (n = 586) were transferred at 96 or 120 h to recipients (n = 49) that had undergone synchronous oocyte recovery as donors. The percentage of recipients producing kittens after transfer of embryos cultured for 96 or 120 h in F-10 was 31 and 25, respectively, compared with 55% of 120 h recipients receiving embryos cultured in M-199 or Tyrode's. Overall, more pregnancies occurred following transfer of > or = 12 embryos (11/26) than if < 12 embryos were transferred (6/23).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8229927 TI - Effect of medium-199 and fetal calf serum on in vitro maturation of domestic cat oocytes. AB - In vitro maturation (IVM) and fertilization are potentially useful for propagating threatened or endangered species. The domestic cat is currently used in this field as an experimental model for studies aimed at non-domestic Felidae. At present optimal conditions for obtaining IVM of cat oocytes have not yet been completely defined. The aim of this study was to evaluate two parameters derived from the procedures that currently ensure high maturation rates in domestic ruminants: (1) the suitability of a complex medium (M-199) for IVM of cat oocytes; (2) the effect of different concentrations of fetal calf serum in the culture medium with or without the addition of gonadotrophins. The maturation rate at two different intervals from the onset of culture (24 and 48 h) was also evaluated. The use of M-199 allowed resumption of meiosis in 4.3-18.7% of cat oocytes, according to the supplements and culture periods used. No significant differences were recorded among the treatment groups (P > 0.05). Meiosis was completed in 90.9% of cases within 24 h with no significant differences between the three treatment groups (P > 0.05). A 3.2% rate of parthenogenesis was observed at the end of the maturation period with no significant differences between the culture systems (P > 0.05). However, the percentage cleavage of oocytes was much higher (29%) when correlated with the percentage that had matured. In this case significant differences among treatments were also observed (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229928 TI - Superovulation and embryo culture in vitro following treatment with ultra-pure follicle-stimulating hormone in cats. AB - Oestrus was induced in nine prepubertal and nine adult cats using three different doses (2.5, 10 and 30 iu divided into five daily doses) of an ultra-pure preparation of porcine follicle-stimulating hormone (FSH) without any luteinizing hormone activity. After 5 days of treatment, ovulation was induced by two daily injections of human chorionic gonadotrophin and the cats were mated. One week later (on day 12) the ovaries were examined for the number of unovulated follicles and ovulation sites. In eight of the nine prepubertal cats the ovaries had 20-90 large follicles per cat and only two cats ovulated (8-9 corpora lutea). All adults responded and ovulated. Adult cats had 3-40 large follicles and 6-35 corpora lutea each. In each group, ovarian response (and ovulation rate) was related to the dose of FSH. The 79 embryos that could be recovered at days 11-13 after the onset of treatment, using a new technique, were assessed; those of good quality were cultured until complete degeneration. One eight-cell embryo was cultured for 12 days through the blastocyst to the gastrula-somite stage. With the highest FSH dosage there was increased degeneration of embryos. This may be associated with the high oestrogen concentrations that were produced during oestrus induction. Potential culture of feline embryos up to the advanced blastocyst stage in modified phosphate-buffered saline supplemented with fetal calf serum was demonstrated. PMID- 8229929 TI - Fertilization in vitro of oocytes matured in vivo in the blue fox (Alopex lagopus). AB - Six unmated blue fox vixens were killed 2-3 days after peak vaginal electrical resistance. Measurements of luteinizing hormone (LH) showed that this corresponded to a time period of 6-10 days after the preovulatory LH peak in these vixens. A total of 63 ova were collected from the oviducts of the vixens, and 45 of the ova were used for study of fertilization in vitro. The ova were placed in 50 microliter droplets of fertilization medium and inseminated with 15,000-45,000 frozen-thawed silver fox spermatozoa that had been separated by swim-up technique, except for nine ova that served as controls for parthenogenesis. Only two of the 36 inseminated ova, both from a vixen killed 6 days after the preovulatory LH peak and inseminated with 15,000 spermatozoa, developed past the four-cell stage. One reached the morula stage 144 h after insemination. Despite a low success rate, the present experiment shows that fox ova matured in vivo may be fertilized and undergo initial embryonic development in vitro. PMID- 8229930 TI - In vitro maturation and fertilization of preovulatory dog oocytes. AB - Mature beagle bitches were injected with exogenous gonadotrophins to induce superovulation. Preovulatory oocytes were collected at 72 h after intravenous injection of human chorionic gonadotrophin and cultured for 72 h in a modified Krebs-Ringer bicarbonate solution containing 10% fetal calf serum and 30 mg gentamicin sulfate l-1. Control oocytes were collected from ovaries of anoestrous bitches and cultured for 72 h. All oocytes were at the germinal vesicle stage when collected. At 72 h of culture, a significantly larger population (31.9%) of preovulatory oocytes reached metaphase II, compared with the control group (12.1%). Ejaculated beagle spermatozoa were washed and preincubated for 4 h, then capacitated spermatozoa were added to the oocytes. The kinetics of sperm penetration into the ooplasm did not differ between the two groups of oocytes, but the development of male and female pronuclei was observed more frequently in oocytes collected from preovulatory follicles. These results suggest that preovulatory maturation is required for the normal development of canine oocytes, although they are ovulated at the germinal vesicle stage. PMID- 8229931 TI - Molecular biological methods for monitoring oocyte maturation and in vitro fertilization in bitches. AB - Oocytes were collected from ovaries of bitches, at various stages of the oestrous cycle, after routine sterilization. Cumulus-enclosed oocytes were cultured for 0 72 h in a modified M-199 medium containing 10% oestrous bitch serum and 20 micrograms oestradiol ml-1. For oocytes surrounded by two or more layers of cumulus cells, an increase in the expression of mRNA transcripts for zona pellucida glycoprotein 3 (ZP3) was seen and reached peak levels after 48 h culture in vitro. Histological assessment showed that 39% of these oocytes had extruded their first polar body after 24 h culture in vitro. When these in vitro matured oocytes were transferred to oviduct cell monolayers and inseminated with fresh dog spermatozoa in Medium-199 supplemented with 10% fetal calf serum and 50 mg gentamicin sulfate ml-1, penetration of the zona pellucida started 1 h after insemination for oocytes that had been cultured for 48 and 72 h. At 12 h after insemination both male and female pronuclei were seen in 37.5% and 20% of the oocytes incubated for 48 and 72 h, respectively. No further development was seen. PMID- 8229932 TI - Cryopreservation of dog semen: a review. AB - Success with artificial insemination of frozen dog semen has been limited. The major problems are associated with identifying the time of optimal fertility for insemination of the bitch, achieving intrauterine deposition of spermatozoa, the variation in post-thaw semen quality between individual dogs and poor spermatozoal survival after freeze-thawing. Many of these problems have been investigated and, to some extent, overcome; however, poor post-thaw spermatozoal longevity remains a problem and results from the lack of ordered logical studies of cryopreservation. The purpose of this paper is to review studies of dog spermatozoal preservation and insemination and to highlight those areas of cryopreservation that have been neglected. PMID- 8229933 TI - Sperm morphology and fertility in the dog. AB - Normal and abnormal dog spermatozoa were studied by means of bright field light and transmission electron microscopy and the effect of abnormal spermatozoa on dog fertility was evaluated. Semen samples were collected by digital manipulation from 67 dogs of 34 different breeds. Ten of the samples were examined ultrastructurally. The association between the percentage abnormal spermatozoa in the ejaculate and the fertility of the dog was statistically evaluated, based on insemination of 42 bitches. On this basis, the male dogs were divided into normal and subnormal groups. The percentage normal morphology below which fertility was adversely affected was found to be 60%: the fertility of dogs with > 60% normal morphology was 61% (14 of 23 inseminated bitches) whereas the fertility of dogs with < 60% normal morphology was 13% (two of 15 inseminated bitches). There was no statistical difference between the ages of the dogs in the two groups; from this it was concluded that subfertility may affect a dog at any age. PMID- 8229934 TI - Hypo-osmotic swelling of dog spermatozoa. AB - Dog spermatozoa were evaluated for their ability to swell when exposed to hypo osmotic conditions. A medium was developed to allow spermatozoal swelling to be identified easily; both the number of swollen spermatozoa and their morphological appearance were influenced by the characteristics of this medium. The number of swollen spermatozoa was inversely proportional to the number of dead spermatozoa in a sample. Although spermatozoal swelling was highly repeatable there was no correlation with spermatozoal motility, morphology or vital staining. Electron microscopic studies demonstrated that swelling of spermatozoa was characterized by ballooning of the tail membrane and internal flexion of the axial filaments. Spermatozoa with damaged membranes were unable to swell. Hypo-osmotic swelling of dog spermatozoa is reliable and repeatable and may be a useful assay for the diagnosis of spermatozoal membrane function. PMID- 8229935 TI - Computer-assisted analysis of motility, velocity and linearity of dog spermatozoa. AB - Two different computer systems were tested for the analysis of motility parameters of dog semen. First, the Cellsoft computer videomicrography system was adapted to the morphological and motility traits of dog semen. The computer calculated motility (percentage of progressively motile spermatozoa) was significantly correlated with the estimated motility (r = 0.97, P < 0.001), as was the sperm concentration determined by counting chamber and by computer (r = 0.62, P < 0.001). Mean velocity varied from 61.1 to 102.0 microns s-1. Irrespective of a low or high motility, the linearity varied from 5.9 +/- 3.5 to 7.6 +/- 0.9. Second, in an investigation into suitability of different concentrations (4, 6 and 8%) of glycerol for freezing dog semen diluted with a Tris-egg-yolk extender, the Stromberg-Mika Cell Motion Analyser was used. Particles identified as spermatozoa ranged in size from 35 to 350 pixels. The mean computer-calculated motility was 93.1 +/- 2.9% in the diluted semen without glycerol; 10 min after adding glycerol it was 88.0 +/- 3.8% with 4% glycerol, 83.7 +/- 4.9% with 6% glycerol, and 78.6 +/- 6.3% with 8% glycerol. Significant correlations were found between the estimated and the computer-analysed motility for the fresh diluted semen (P < 0.01) and for the thawed semen, irrespective of the glycerol concentration (P < 0.001). The mean velocity was 159.3 +/- 13.8 microns s-1 before adding glycerol, showing a decreasing tendency thereafter.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229936 TI - Fine structure and elemental composition of fresh and frozen dog spermatozoa. AB - Dog spermatozoa from fresh ejaculates and after freezing-thawing were air-dried on to grids and subjected to X-ray microanalysis or examined by scanning and transmission electron microscopy. The spermatozoa subjected to freezing-thawing presented major changes in their morphology, which included the loss of acrosomal contents, seen both by the swelling and rarefaction of the acrosome and the loss of electron-dense acrosomal material. In many cells the acrosomal damage, including the equatorial region, was conspicuous, with vesiculization of the acrosomal membranes. The plasmalemma remained, in most cases, apparently intact. X-ray microanalysis of the sperm aliquots showed a significant decrease in the amount of most selected elements after freezing-thawing. The extent to which concentrations of phosphorus and sulfur decreased in the sperm head region suggested that major changes in the chromatin occurred during the freeze-thaw process. PMID- 8229937 TI - Sperm abnormalities in silver fox (Vulpes vulpes) semen selected for artificial insemination. AB - Sperm morphology was examined in fresh ejaculates accepted for insemination. The material included 161 ejaculates of 36 male silver foxes. Samples were fixed, stained with eosin/nigrosin and evaluated microscopically. The abnormal spermatozoa were categorized by the location of the defect. The mean (+/- SD) percentage of morphologically normal spermatozoa was 87.55 +/- 10.45. The incidence of sperm defects was low and variation high in all defect classes. Defects of tails of spermatozoa accounted for > 50% of the abnormalities found. Morphological quality did not correlate with the volume of the sperm-rich fraction, mass activity, progressive motility, or total sperm count in ejaculates of acceptable sperm quality. The incidence and effects of morphological defects in low-quality semen remain to be studied. PMID- 8229938 TI - Biology of ovarian cycles, pregnancy and pseudopregnancy in the domestic cat. AB - Puberty in the female cat occurs between the ages of 8 and 10 months. Cats are seasonally polyoestrous, reflex ovulators. The oestrous cycle can occur as often as every 2-3 weeks. Ovulation usually occurs 24-36 h after copulation, and implantation occurs 12-13 days after copulation. The duration of gestation in the cat is 64-67 days (average 66 days). The corpora lutea secrete increasing amounts of progesterone, starting 1-2 days after ovulation. If implantation occurs, progesterone concentrations continue to increase throughout days 25-30, then slowly decline throughout the rest of pregnancy. In the absence of pregnancy, the corpora lutea reach their peak progestational activity within 10-15 days and then decline, with basal progesterone values being noted by days 30-35. Relaxin is produced by the fetoplacental unit beginning at about day 20 of gestation and continuing throughout the rest of pregnancy. Prolactin production increases from about day 35. Like those of prostaglandin F2 alpha, concentrations of prolactin plateau at about day 50 and increase abruptly just before delivery. PMID- 8229939 TI - Effects of four different extenders and three different freezing rates on post thaw viability of dog semen. AB - Ejaculates were collected from 17 dogs. After initial evaluation the ejaculates were split into four equal aliquots and diluted with either Triladyl, Pipes, IMV Universal or Tris-fructose-citric acid extender, all containing egg yolk and glycerol. The extended semen was frozen in 0.5 ml straws at a slow, intermediate or fast freezing rate. After thawing, the percentage of progressively motile spermatozoa and the velocity of forward motion was evaluated. There was no difference between the extenders with regard to progressive motility. The slow, intermediate and fast freezing rates resulted in the highest, intermediate and lowest motility values, respectively, regardless of extender. All four extenders were equally suitable for freezing canine semen at a slow freezing rate. PMID- 8229940 TI - Comparative cryopreservation and capacitation of spermatozoa from epididymides and vasa deferentia of the domestic cat. AB - Methods of cryopreservation for spermatozoa from domestic cat epididymides and vasa deferentia were compared as models for posthumous gamete salvage from non domestic felids. Spermatozoa were collected either immediately after castration (Fresh, n = 37) or after being cooled (5 degrees C) in tissue overnight (Cool, n = 37) and released into one of three extenders containing 20% egg yolk and 3% glycerol for cryopreservation: (1) TE: Tris buffer, citric acid and fructose; (2) TC: Tris buffer, citric acid and glucose, or (3) CP: lactose, and frozen over lipid nitrogen. Before and after freezing, each sperm cell sample was evaluated for motility and percentage morphologically normal cells. Samples were also evaluated for their ability to initiate fertilization using a zona attachment assay. Neither percentage morphologically normal spermatozoa nor percentage motility differed among the three diluents for prefreeze and post-thaw samples, regardless of the collection treatment. However, CP tended to provide lower post thaw status than did the TE and TC cryoextenders. Before freezing, there was no difference in percentage motility between the Fresh and Cool groups (mean: 76 versus 72%, respectively); however, progressive status and normal morphology were lower (P < 0.05) in Cool (3.0 and 57%) than in Fresh (3.4 and 64%) samples. After thawing there was a greater decline (P < 0.05) in percentage motility in the Cool than in the Fresh group (34 versus 24%) and the number of intact acrosomes dropped from prefreeze values of 66.7 +/- 6.3 and 56.4 +/- 4.8% to 17.8 +/- 3.9 and 20.9 +/- 4.6% after thawing in the Fresh and Cool groups, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229941 TI - Biology of gonadotrophin secretion in adult and prepubertal female dogs. AB - Studies in the female domestic dog demonstrate that luteinizing hormone (LH) and follicle-stimulating hormone (FSH) have secretion patterns that are pulsatile, are inhibited by oestradiol during pro-oestrus and surge to maximal values before ovulation. Studies in ovariectomized bitches suggest that the periovulatory surge is triggered by a preovulatory decline at late pro-oestrus in the oestrogen:progesterone ratio. During the 3-8-month non-seasonal anoestrus, FSH concentrations usually are 50-100% of those of the periovulatory peak, whereas LH concentrations are only 10-20% of peak values. In ovariectomized bitches FSH concentrations are often 5-10 times preovulatory peak values, whereas LH concentrations are only the same as, or double, peak values. Increased LH concentration and pulse frequency are associated with the termination of anoestrus. Treatment with gonadotrophin-releasing hormone (GnRH) pulses or infusions of GnRH agonists can induce fertile oestrus in early anoestrous bitches, as can treatment with a dopamine agonist, presumably by suppression of prolactin secretion. Between 4 months of age and pubertal pro-oestrus at 8-12 months of age, serum concentrations of FSH and LH are similar to those in adult anoestrus, and are suppressed during chronic infusion of GnRH agonist. The latter resulted in a reversible inhibition of puberty during 1 year of treatment. Studies in vivo have shown that LH and prolactin are luteotrophic throughout most of the luteal phase. LH stimulated progesterone synthesis by bitch luteal cells in vitro in the presence or absence of stimulant factors or steroid precursors present in serum. PMID- 8229942 TI - Non-surgical intrauterine artificial insemination in bitches using frozen semen. AB - A total of 46 bitches were inseminated directly into the uterus using non surgical insemination procedures; the technique used in six bitches involved specially designed metal catheters and abdominal fixation of the cervix, whereas the remainder were inseminated by passing a flexible plastic catheter through the cervix using direct endoscopic visualization to facilitate the process. Twenty seven bitches were inseminated with semen frozen at the clinic; the remainder were inseminated with imported semen. Insemination timing was based on endoscopic assessment of the vaginal mucosa, vaginal cytology and blood progesterone concentration determined using a rapid, qualitative enzyme-linked immunosorbent assay (ELISA) kit. Each bitch received between 50 x 10(6) and 200 x 10(6) total spermatozoa per insemination; post-thaw motility varied from 10 to 80%. Two inseminations were performed 48 h apart in the majority of bitches. An overall pregnancy rate of 80% (37/46) was obtained with a mean litter size of 5 +/- 3.14. Subsequent pregnancy rates were comparable for both techniques and both were considered to be effective methods of inseminating frozen semen. Considerably fewer spermatozoa were inseminated in many of these bitches than have previously been reported. In a series of seven bitches using the semen from one dog, each bitch received two inseminations of 30-35 x 10(6) live normal spermatozoa per insemination. A pregnancy rate of 85% (6/7) and a mean litter size of 7.8 was achieved. Rapid ELISA progesterone kits were used to identify the optimum time for insemination. PMID- 8229943 TI - Results of 527 controlled artificial inseminations in dogs. AB - During 1990 and 1991, 527 artificial insemination (AI) procedures were performed by 40 veterinarians, under the Swedish Kennel Club control scheme, and reported to the Kennel Club within 14 days. The most common reason for AI (37.2%) was import of fresh and frozen semen or quarantine, i.e. introduction of new genetic material. Fresh semen (468 AIs) was deposited into the cranial vagina, and frozen semen (59 AIs) was inseminated transcervically into the uterus. Pregnancy rates were 54.7% with fresh and 39.0% with frozen semen. When corrected for stage of oestrus at the time of AI and semen quality, the pregnancy rates were 62.3 and 51.1%, respectively. Bitches inseminated with frozen semen had a 29.7% lower pregnancy rate and 30.5% smaller litter size than bitches receiving fresh semen; these differences were statistically significant. Forty-four bitches (8.4%) were also mated. Their pregnancy rate was significantly higher (88.6%; 90.7% corrected) than that of bitches inseminated with fresh semen only. Pregnancy rate was significantly higher in bitches inseminated twice with fresh semen than in those inseminated once only. Litter size and gestation length increased with the number of AIs. Most bitches whelped 62 days after a single AI with fresh or frozen semen, but the gestation period varied between 55 and 69 days. There were significant differences in fertility between breeds. Pup deaths occurred in 33.8% of the litters and the pup death rate during the first 3 weeks of life was 11.6%. The number of pups resulting from AI amounted to 1.1% of the total number of pups registered with the Swedish Kennel Club. PMID- 8229944 TI - Canine artificial insemination with frozen semen: comparison of intravaginal and intrauterine deposition of semen. AB - A total of 57 bitches of various breeds were inseminated with frozen semen [the extender used being Tris-fructose-egg yolk-6.4% (v/v) glycerol] under controlled conditions. The optimum time of insemination was determined using vaginal smears and progesterone assays. Of 38 bitches inseminated one to three times in the vagina using a technique employing a commercial intravaginal pipette, 20 (52.6%) became pregnant, with a mean litter size of 4.2. Of 19 bitches inseminated one to three times using a transcervical technique, the conception rate was 73.6% with a mean litter size of 5.5. These data show, for the freezing method used, that the results of artificial insemination with frozen semen in the bitch are better with intrauterine than with intravaginal deposition. PMID- 8229945 TI - Effect of addition of autologous prostatic fluid on the fertility of frozen thawed dog semen after intravaginal insemination. AB - Vaginal insemination of frozen-thawed dog semen usually gives a highly unpredictable fertility rate with poor pregnancy rates in most bitches. This study was designed to establish whether litter size, pregnancy rate and conception rate could be improved by the addition of autologous prostatic fluid to the frozen-thawed semen before insemination. Twenty German shepherd bitches that were free of any clinical reproductive abnormality or suspect breeding history were used. The bitches were stratified according to their ages and randomly assigned within strata to treatment (group T, n = 10) or control groups (group C, n = 10). All bitches were inseminated daily with frozen-thawed semen for the duration of that stage of vaginoscopic oestrus during which the vaginal folds were shrunken and angular. Bitches in group T were inseminated with semen to which 7-10 ml of frozen-thawed sperm-free autologous prostatic fluid has been added immediately before insemination. No prostatic fluid was added to the semen used to inseminate the bitches in group C. Each inseminate contained 100 x 10(6) progressively motile spermatozoa after thawing. Semen was deposited in the vaginal fornix using a disposable plastic bovine artificial insemination pipette. For groups T and C the mean number of conceptuses per cycle (litter size), the mean ratio of conceptuses to corpora lutea (conception rate), and the pregnancy rate were 5.2 +/- 3.01 and 2.4 +/- 2.84, 0.577 +/- 0.35 and 0.23 +/- 0.27, and 100% and 60%, respectively. The addition of autologous prostatic fluid to frozen thawed canine spermatozoa significantly improved the litter size (P = 0.023), the conception rate (P = 0.0127) and pregnancy rate in bitches (P = 0.043).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229946 TI - Response to gonadotrophin-releasing hormone by the intact male dog: serum testosterone, luteinizing hormone and follicle-stimulating hormone. AB - In Expt 1, gonadotropin-releasing hormone (GnRH) (Cystorelin, CEVA) was administered intramuscularly to two intact male dogs; one dog received one injection of 50 micrograms GnRH and one dog received four daily injections of 50 micrograms GnRH. Both dogs exhibited a significant and immediate rise in luteinizing hormone (LH) following GnRH administration, with a peak observed at 15 min following injection. Testosterone was increased over baseline concentrations for 5 and 7 days, respectively, after the injection. In Expt 2, eight intact male dogs were injected intramuscularly with 0.7 microgram GnRH (Factrel, Fort Dodge) kg-1. Baseline testosterone concentrations were established by daily sampling for 14 days before treatment. All dogs exhibited an LH peak 15 min and a testosterone peak 60 min after the GnRH injection. Testosterone concentrations had returned to baseline concentrations by 4 h after the injection. Testosterone tended to fall below baseline concentrations for several days following the injection of GnRH. No peak was noted for follicle-stimulating hormone. In Expt 3, five additional dogs were injected with 0.7 microgram GnRH (Factrel, Fort Dodge) kg-1. Testosterone concentrations rose in all dogs 1 h after the injection and returned to baseline concentrations by 24 h after injection. In the male dog, GnRH stimulated an LH peak 15 min and a testosterone peak 1 h after injection. Further investigations are needed to elucidate the different effects on testosterone concentration observed with two different GnRH preparations. PMID- 8229947 TI - Postnatal development of seminiferous tubules in the cat. AB - Testes were obtained from 20 cats with an age range of 1 day to 1 year. In the neonatal period the seminiferous tubules were widely separated by a well developed interstitium. Tubular diameter was small and no lumen was observed within the seminiferous tubules. These showed many undifferentiated Sertoli cells with round to ovoid nuclei and only one or two spermatogonia, usually centrally situated. From 2 to 4-5 months the interstitium was sparse, tubular diameters were increased and the morphology of the seminiferous tubules was very similar to that in the neonatal period (< 2 months). From 5 to 7 months, tubular diameters were larger, a lumen was observed, the seminiferous epithelium began to assume its characteristic stratification and the blood testis barrier was developed. From 8 months of age spermatogenesis was fully established. PMID- 8229948 TI - Histological study of Leydig cells in the cat from birth to sexual maturity. AB - Testes of 20 cats with an age ranging from 1 day to 1 year were obtained. Histological study revealed three different populations of Leydig cells. In the early postnatal period (from birth to 1 month) they were round or polyhedral with an ovoid and eccentric nucleus. From 2 to 4-5 months Leydig cells were similar to those seen in the early postnatal period, showing shrunken nuclei with condensed chromatin masses and accumulation of lipid droplets in the cytoplasm. The peritubular Leydig cells mostly contained elongated nuclei. From 5 to 6 months of age adult Leydig cells were encountered, generally polygonal in outline with large nuclei that were usually round to ovoid and displaced to one side of the cell. The cytoplasm contained lipid droplets and lipofuscin granules. The number of testosterone-containing cells decreased progressively from 1 to 4 months; from 3 to 4 months only a small number of testosterone-containing cells were present. PMID- 8229949 TI - Vaginal anatomy of the domestic cat (Felis catus) in relation to copulation and artificial insemination. AB - In the course of an investigation of artificial insemination (AI) in the domestic cat, the site of deposition of the semen during vulval insemination was questioned. A limited study was carried out to ascertain both the anatomy and distensibility of the vagina and urogenital sinus, and the location of the cervix in relation to the vulva. In live restrained female cats a 4 mm diameter Teflon probe could not be inserted more than 20 mm, and elicited a cry similar to the copulatory vocalization when the resistance was encountered. However, a 1 mm diameter ball-tipped needle was passed to a depth of 40 mm. To confirm these observations, four adult female domestic cats were dissected. The cervix was located approximately 45 mm cranial to the vulva. The urogenital sinus narrowed approximately 17.5 mm from the vulva, preventing the passage of a 4 mm probe, and led to a constricted vagina, which permitted the passage of a 1 mm diameter probe to the depth of the cervix. A radiological investigation of a single anaesthetized cat supported these results. The erect penis of the adult male was measured during sexual arousal in five animals; the mean length and width (+/- SD) was 21.2 +/- 2.2 mm and 5.1 +/- 0.5 mm, respectively. These observations lead us to conclude that the stimulus for vocalization during natural mating is distention of the posterior vagina, where semen is probably deposited. The site of semen deposition is uncertain in several accounts of artificial insemination, as the passage of the insemination device depends on its diameter. PMID- 8229950 TI - Comparison of induction of oestrus in dogs using metergoline, metergoline plus human chorionic gonadotrophin, or pregnant mares' serum gonadotrophin. AB - Twenty beagle bitches were treated with metergoline (groups I and II), ten with pregnant mares' serum gonadotrophin (PMSG; group III) and ten served as an untreated control group. Metergoline (12.5 mg per bitch) was injected intramuscularly (i.m.) every 3 days, starting between days 78 and 161 after the previous pro-oestrus. In 18 of the metergoline-treated bitches the interpro oestrous interval (144 +/- 29 days) was significantly shorter (P < 0.01) than that of the control group (206.7 +/- 29.7 days). Of these 18 responder bitches, ten had not received human chorionic gonadotrophin (hCG) whereas eight had received 500 iu hCG i.m. in late pro-oestrus (group II). The durations of pro oestrus and oestrus of the ten responding bitches not given hCG (6.8 +/- 5.5 and 11.7 +/- 4.5 days, respectively) did not differ (P > 0.05) from those of the control group (7.8 +/- 3.6 and 11.2 +/- 3.2 days). All ten bitches ovulated and nine became pregnant. Two of eight bitches responding to metergoline in group II exhibited a prolonged pro-oestrus after injection of hCG. In the remaining six bitches, the average duration of pro-oestrus and oestrus was 4.8 +/- 1.7 and 6.5 +/- 2.8 days, respectively, which was significantly shorter (P < 0.05) than in bitches not given hCG. The incidences of ovulation (6/8) and gestation (4/8) were lower than in bitches that received metergoline alone, or in the control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229951 TI - Effect of body condition and dietary energy supply on reproductive processes in the female mink (Mustela vison). AB - This paper reviews a series of experiments regarding the influence of body condition and dietary energy supply on reproductive processes in the female mink. The reporting is based on 1905 sets of reproductive performance data and on data regarding number of corpora lutea, implantation rate, plasma progesterone and oestradiol, luteinizing hormone (LH) release and ovulation rate, and the early development of the fertilized ova. Generally, treatment effects were more pronounced in yearling females than in adults; furthermore, adults responded less to varied energy supply by changes in body weight. A weight reduction of > 300 g during the winter resulted in poor reproductive performance in yearling females. A long-term study indicated better reproductive longevity for females on a low intensity feeding regimen, whereas females on a high-intensity feeding regimen tended to have larger litters until 3 years of age. Nutritional flushing was carried out by a 2-week period of moderate energy restriction followed by feeding ad libitum from 3-5 days before initiation of matings until matings were completed for the individual females. Mainly in yearlings, but also in adults, litter sizes were improved for females with a first mating early in the breeding season, which was confirmed by an increased number of corpora lutea. The interval between the first recorded increase in plasma progesterone and the day of the peak progesterone concentration was decreased. Plasma oestradiol concentrations were higher in flush-fed females; age-dependent differences in the appearance of the profiles were also found. LH release tended to be more synchronized and distinct in flush-fed females.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229952 TI - Effect of hyperadrenocorticism and diabetes mellitus on serum progesterone concentrations during early metoestrus of pregnant and non-pregnant cycles induced by pregnant mares' serum gonadotrophin in domestic dogs. AB - The intramuscular (i.m.) administration of pregnant mares' serum gonadotrophin (PMSG) (20 iu kg-1 day-1 for 5 days) followed by an injection of human chorionic gonadotrophin (500 iu, i.m.) induced oestrus in 28 cross-bred bitches. These included, based on endocrine evaluations, nine normal (N), seven insulin deficient diabetic (ID), six insulin-resistant (IR) and six hypercortisolic (HC) bitches. The bitches were not bred but were allowed to have a non-pregnant cycle. When oestrus was induced a second time by re-treating at 35 days after the end of the non-pregnant metoestrus of the first induced cycle, the dogs were mated. Pregnancy was obtained in five N, four ID, four IR and three HC dogs; 15 of the 16 dogs had litters. For these 16 dogs, progesterone concentrations in serum were evaluated from samples that had been obtained daily during the first 30 days of the first (non-pregnant) induced cycle and the first 11 days of metoestrus of the second induced (pregnant) cycle. Corpus luteum development was characterized in normal bitches by a steady increase in progesterone, with highest values (33 +/- 6 ng ml-1) at day 8 of metoestrus in pregnant dogs. Progesterone averaged among the first 11 days of metoestrus (pregnant and non-pregnant) did not differ between normal and insulin-resistant bitches (20 +/- 5 versus 22 +/- 4 ng ml 1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229953 TI - Induction of follicular development in silver foxes (Vulpes vulpes) with equine chorionic gonadotrophin (eCG) and antibodies against eCG. AB - Experiments were conducted that demonstrated that 1000 iu equine chorionic gonadotrophin (eCG) was effective in induction of follicular development and ovulation in silver foxes during anoestrus. This treatment resulted in large, unovulated follicles; thus, trials in which the effects of eCG were reduced or abrogated by antibodies against eCG were carried out. Passive immunization against eCG on days 3 and 4 after eCG treatment interfered with subsequent follicular development and prevented ovulation. Treatment with eCG antibodies on days 5 or 7 after eCG treatment did not prevent ovulation and neutralization beginning on day 5 appeared to provide for the best ovulatory yield. The results suggest that combinations of eCG and anti-eCG antibody may provide a useful means of inducing ovarian activity in anoestrous foxes. PMID- 8229954 TI - Effects of medroxyprogesterone acetate or gonadotrophin-releasing hormone agonist on suppression of spermatogenesis in the dog (Canis familiaris). AB - Medroxyprogesterone acetate (MPA) was administered as a depot injection to two adult male dogs. One was injected with 75 mg as a depot injection once a month for 2 months and the other with 50 mg at 2-weekly intervals for 1 month, followed by 100 mg at 2-weekly intervals for a further 2 months. Two other male beagles were given gonadotrophin-releasing hormone (GnRH) agonists (nafarelin derivative no. 49947 and nafarelin acetate) at a dose of 50 micrograms day-1 for either 22 or 23 days. The profiles of luteinizing hormone (LH) and testosterone were monitored before and during the treatment periods, and changes in the dogs' testes, epididymides and prostate glands were determined post mortem. In the MPA treated animals, LH was suppressed for only very brief periods. In contrast, in the dogs treated with GnRH agonist, LH and testosterone concentrations were suppressed for most of the treatment period. In the MPA-treated animals, histological changes were seen mainly in the head of the epididymis, and in the acrosome region of the spermatozoa still present in this region of the male reproductive tract. The acrosome, instead of being closely applied to the sperm head, was in contact only in the most anterior areas, with the remainder forming a fan-like projection, indicating increasing detachment from the sperm head. In the dogs treated with GnRH agonist there was clear evidence of cessation of normal spermatogenesis in most of the seminiferous tubules. Changes in the histology of the head of the epididymis were similar to (but more profound than) those found in the dogs treated with MPA. Seminal changes were demonstrated in ejaculates collected during the latter stages of the treatment period. PMID- 8229955 TI - Laparoscopic vasectomy in the male dog. AB - A laparoscopy technique was employed to sterilize five adult male dogs. The animals were sedated with a combination of medetomidine and buprenorphine intravenously and a fragment of the ductus deferens, about 2 cm in length, was removed after having been cauterized and cut, using atraumatic forceps and a pair of scissors connected to an electrocautery unit. The procedure was easy, quick and safe. The animals had no complications after the intervention and made a good recovery. PMID- 8229956 TI - Abortion induction in the cat using prostaglandin F2 alpha and a new anti prolactinic agent, cabergoline. AB - This study in cats compared the effects of a natural prostaglandin F2 alpha (PGF2 alpha) and cabergoline, administered on and after day 30 after mating, with regard to the plasma progesterone concentration, the induction of abortion and the side effects of each regimen. PGF2 alpha, through a direct luteolytic action, induced abortion by an abrupt and rapid (24 h) decline in the plasma progesterone concentration. Using doses (2 mg per cat) comparable to the ones selected in a previous study, 100% of abortions (4/4) in cats treated from day 33 of gestation were obtained. Cabergoline (1.65 micrograms kg-1 day-1, administered subcutaneously for 5 days), through its antiprolactinic action induced abortion in 80% (4/5) of the cats treated on day 30 of gestation. The abortion was initiated by means of a reduction in plasma progesterone concentration to < 1 ng ml-1. This reduction was not as rapid (3-4 days), however, as that obtained with PGF2 alpha (24 h). Prostaglandins always induced significant side effects such as nausea, prostration, vomiting and diarrhoea, within 10 min following injection, whereas cabergoline never induced side effects or behavioural disturbances. In addition, cabergoline usually induced abortion through fetal resorption (75% of cases), without any clinical sign other than some vaginal discharge. PMID- 8229957 TI - Reproduction in a feral cat population and its control with a prolactin inhibitor, cabergoline. AB - A feral cat population (12 females, 13 males) was observed almost daily for 6 years, at a location 40.53 degrees N and 74.29 degrees W. Cats were accustomed to twice-daily feeding. Oestrous behaviour was seasonal and started in early January, irrespective of climatic conditions, with peak levels of oestrous activities in late January and February. Of all 60 heat periods observed, and conceptions recorded, the percentage occurrence was 58 and 49 in the first quarter, and 29 and 33, 12 and 16 and 1 and 2 in the second, third and fourth quarter, respectively. Queens could be reliably identified as being pregnant by 31.5 +/- 5.82 days, and were subjected between days 36 and 57 of gestation to daily oral treatment with 5-15 micrograms cabergoline kg-1 placed on food. Treatments (n = 41) from days 36 +/- 6.17 to 40.8 +/- 6.96 resulted in abortion in all animals on day 40.5 +/- 6.19; if treatment started as late as day 48.5, and lasted 9 or more days, premature parturition occurred. As cabergoline had caused mammary gland regression within 36-48 h, litters could not be nursed and perished quickly. Five repeatedly aborted queens were subsequently allowed to go to term. Pregnancies, gestation length (64.5 days), parturitions, maternal care and lactations were normal. Of the 24 kittens born out of seven pregnancies, 16 were observed from 6 months to 3 years. Pubertal oestrus and first conceptions occurred at 189.9 days (range 150-214 days) and 212.0 days (155-277 days), respectively. Males were seen to achieve reproductive success in their third year only. PMID- 8229958 TI - Clinical use of prostaglandin F2 alpha to induce early abortion in bitches: serum progesterone, treatment outcome and interval to subsequent oestrus. AB - Records of elective abortion in 27 bitches presented to the Veterinary Surgery and Reproduction Clinic of the University of Pisa were reviewed. Treatment consisted of twice-daily subcutaneous injections of 150-270 micrograms natural prostaglandin F2 alpha kg-1 for 4 consecutive days starting between cytological dioestrous days 5 and 19. None of the treated bitches whelped. Luteolysis appeared to be complete (serum progesterone < 3.2 nmol l-1) at the end of treatment in 22 of 25 bitches in which progesterone was assayed in serum obtained daily during therapy. Serum progesterone rose after treatment to a concentration consistent with pregnancy in four of 17 bitches in which progesterone was measured once between dioestrous days 23 and 51. The mean intercycle interval (pro-oestrus-pro-oestrus) of the cycle in which abortion was induced was 134 +/- 45 days for 15 dogs for which the interval could be determined, and was significantly shorter than the average preceding interoestrous intervals (210 +/- 40.0 days) for the same dogs (P < 0.001). PMID- 8229959 TI - Pet (dog and cat) overpopulation in the United States. AB - Over half of all United States (US) households own a dog or cat. The veterinary profession can now provide health care for dogs and cats of affluent or devoted owners that rivals the health care offered many human patients. Unfortunately, as many pets receive medical and surgical care that becomes increasingly sophisticated, other pets in the US receive no veterinary care at all. Additionally, millions of pets are humanely killed in US animal shelters because owners are not committed to the continual responsibilities of pet care. Although the total dog and cat population is unknown in the US, as is the total number of pets killed, estimates suggest that between one-tenth and one-quarter of the entire US pet population is destroyed annually because of a surplus dog and cat problem. Pet overpopulation is attributable to relinquishment and abandonment, as well as to birth rates; thus, veterinarians must strive to reduce pet overpopulation by not only curbing reproduction, but also by decreasing the major cause of pet death in the US (i.e. humane killing). Thus, the veterinary profession must take a prominent role in the campaign to prevent the deaths of healthy animals for whom homes cannot be found, just as it has done to prevent the deaths of sick animals that do have homes. PMID- 8229960 TI - Genetics of sexual differentiation and anomalies in dogs and cats. AB - Normal mammalian sexual differentiation is dependent upon the successful completion of a series of steps that are under genetic control. These steps are marked by three consecutive events-establishment of chromosomal sex, gonadal sex and phenotypic sex. Chromosomal sex is normally established at fertilization. A gene located on the Y chromosome (Tdy) encodes a protein, the testis-determining factor, that is the genetic switch for male development. Testicular development establishes male gonadal sex. Two testicular secretions are responsible for masculinization of the phenotype: Mullerian inhibiting substance causes the Mullerian duct system to regress; testosterone allows development of the vas deferens and epididymides from the Wolffian ducts. Testosterone is metabolized to dihydrotestosterone in cells of the urogenital sinus, genital tubercle, and genital swellings, stimulating formation of the prostate and urethra, the penis, and the scrotum, respectively. In the absence of the Y chromosome and the Tdy gene, the default pathway to female gonadal sex is initiated. The gonadal anlagen develops into an ovary, establishing female gonadal and phenotypic sex. Abnormalities in sexual differentiation are classified by the initial step, as far as is known, at which development differs from normal. Anomalies are categorized as errors in chromosomal, gonadal or phenotypic sex. Reported abnormalities of genetic sex in the dog and cat include abnormalities in chromosomal number or structure, such as the XXY and XO syndromes, chimaeras and mosaics. Abnormalities of gonadal sex include XX sex reversal in the dog, which is inherited as an autosomal recessive trait with phenotypic expression limited to dogs with an XX chromosome constitution. Abnormalities of phenotypic sex include testicular feminization syndrome (TFM) in the cat and persistent Mullerian duct syndrome (PMDS) in the dog. In complete testicular feminization, the androgen receptor is absent or non-functional. Affected individuals have a normal male karyotype (XY) and bilateral testes, but androgen-dependent masculinization is completely absent. As in other species, TFM in the cat is likely to be inherited as an X-linked trait. PMDS in the miniature schnauzer is inherited as an autosomal recessive trait with expression limited to homozygous males. PMDS-affected male dogs have a normal male karyotype (78,XY), bilateral testes, and a complete Mullerian duct system (oviducts, uterus, cervix and cranial vagina). Current studies support the hypothesis that target organ resistance, possibly a mutation in the gene for the MIS receptor, is responsible. PMID- 8229961 TI - Anovulatory ovarian cycles in dogs. AB - In 14 of 1152 female dogs, monitored and prepared for mating during the last 2 years, a functional disturbance of the ovaries on the basis of vaginal inspection, cytological investigation of vaginal smear and progesterone concentration was observed, manifested by anovulatory sexual cycles. After a normal pro-oestrus of approximately 8-16 days the bitches showed an increase in progesterone concentration to 6.4-9.5 nmol l-1 and in two cases 11.0 nmol l-1, followed by a sudden decrease to the basic rate of 0.64 nmol l-1, associated with all clinical signs indicative of the regression of oestrus. These findings suggest that reliable identification of mating time in female dogs requires monitoring of rising progesterone concentrations up to at least 32.0 nmol l-1. PMID- 8229962 TI - Pregnancy failure in cats associated with long-term dietary taurine insufficiency. AB - The effect of long-term dietary taurine insufficiency on reproductive function was studied in adult female domestic cats (n = 11). Cats were time-mated during taurine-deficient (6 months) and refed (6 months) states, and the outcome of ovulatory cycles and breeding was analysed. Serum progesterone and relaxin concentrations were evaluated in order to characterize pregnancies, including those resulting in resorption of fetuses, and pseudopregnancies. Increased resorption of fetuses, reduced litter size, and increased incidence of stillborn kittens was observed in queens while on taurine-deficient diets, as well as after refeeding of a taurine-enriched diet. Overall, 30% of the ovulatory cycles resulted in the delivery of kittens, with mean live and stillborn litter sizes of 2.2 +/- 0.4 and 0.8 +/- 0.4 kittens (mean +/- SEM), respectively. The remaining ovulatory cycles resulted either in pregnancies in which fetuses were resorbed (38%), or in pseudopregnancies (32%). Ovulatory cycles resulting in resorbed fetuses were characterized by the appearance of relaxin on day 20 of gestation, but with a subsequent decrease to non-pregnant concentrations by day 25 of gestation. These results suggest that reproductive failure in domestic cats exposed to long-term nutritional taurine deficiency is associated with a postovulatory defect manifest within the first 10 days after implantation, and that this defect is not reversible upon refeeding of a taurine-enriched diet for 6 months. PMID- 8229963 TI - Observations on the treatment and diagnosis of open pyometra in the bitch (Canis familiaris). AB - Eight bitches presenting with vaginal discharges were diagnosed as having open pyometra, as assessed by ultrasonographic, radiographic, haematological and clinical investigation. Plasma progesterone was assessed before treatment with prostaglandin F2 alpha (Dinoprost) and three of the bitches had low concentrations. Each bitch was given multiple injections of PGF2 alpha and monitored during and after treatment. All bitches had basal progesterone concentrations after treatment and the uterine diameters were markedly reduced by that time. One bitch has subsequently produced a litter of seven pups. Five of six bitches that were subjected to ovariohysterectomy 1-2 months later had normal uteri, although one had caseated areas in the uterine wall. PMID- 8229964 TI - Relationship between gonadotrophins, inhibin and sex steroid secretion during the periovulatory period and the luteal phase in the blue fox (Alopex lagopus). AB - The blue fox is a monestrous seasonal breeder with a pro-oestrus and oestrus lasting several days and a prolonged luteal phase in non-pregnant as well as pregnant vixens. Ovulation occurs 2 days after the preovulatory luteinizing hormone (LH) peak at the onset of oestrus; oocytes are ovulated as primary oocytes and maturation is completed 2-3 days later. Fertilization is possible during all stages of oocyte maturation until degeneration of oocytes 5 or 6 days after the LH peak. In this study, changes in plasma concentrations of follicle stimulating hormone (FSH) and immunoreactive inhibin (iINH) and their relationships with LH, oestradiol, androstenedione, testosterone and progesterone secretion and with the periovulatory events, were determined throughout the reproductive cycle of 14 vixens. In late anoestrus and pro-oestrus, FSH and iINH were inversely related to each other (r = -0.84, P < 0.001); whereas FSH concentrations declined to reach low values within the 5 days preceding the LH peak, iINH secretion increased, as did that of oestradiol and androgens. Thereafter, iINH and steroid concentrations with the exception of progesterone reached maximal values at the time of the LH and FSH peaks and decreased within the following 2-3 days in the early luteal phase. Progesterone started to increase simultaneously with the LH peak, and when oocyte maturation was completed (around day 4), progesterone concentrations were increasing, while those of other hormones were low. A postovulatory release of FSH was detected between days 6 and 10; it preceded the rise in iINH, which began on day 8 when concentrations of progesterone were already high and those of oestradiol and LH were low. Progesterone and iINH were positively correlated over the period day 6 to day 49 (r = 0.62, P < 0.01). These results indicate that, in the blue fox, iINH is secreted by the preovulatory follicle and the corpus luteum; that during pro-oestrus, iINH may act in synergy with oestradiol to reduce the secretion of FSH and that after the LH peak the decrease of iINH and oestradiol concentrations may be responsible for the postovulatory release of FSH, which in turn causes an increase in iINH, possibly secreted by the corpus luteum and, as a result, FSH secretion is suppressed. PMID- 8229965 TI - Diagnosis and therapy of the subinvolution of placental sites in the bitch. AB - Within the last 19 years, 20 cases of placentitis post partum have been studied at the clinic. On 14 occasions the condition took the form of excessive uterine bleeding (metrorrhagia) post partum and, on five occasions, of prolonged vaginal discharge lasting 8-13 weeks post partum. One bitch with a clinically undetectable form of this disorder was diagnosed at ovariohysterectomy. The average age of the bitches was 4.5 years (range 2-6 years). Examinations included abdominal palpation, vaginoscopy, vaginal smear examination and, if necessary, radiography and ultrasonography. The presence of decidua-like cells in the smear was confirmed as evidence of abnormal uterine involution. Six dogs were subjected to ovariohysterectomy; histological examination revealed inflamed and degenerated areas of the uterine surface. Decidualike cells were seen for up to 10 weeks post partum. Fourteen bitches were treated successfully with an injection of the progestagen Niagestin (25-50 mg per bitch) and both intrauterine and parenteral antibiotics were administered. The discharge stopped within a few days in all dogs treated. A subsequent normal pregnancy occurred in four cases. PMID- 8229966 TI - Lack of effect of trimethoprim and sulfadiazine in combination in mid- to late gestation on thyroid function in neonatal dogs. AB - Four pregnant bitches were randomly selected and were given 25 mg sulfadiazine kg 1 and 5 mg trimethoprim kg-1 orally once daily, starting on day 29 of dioestrus until parturition. Five untreated pregnant bitches were used as controls. At the end of the treatment period (parturition = week 0), one to three pups of each litter, treatment and control group, were randomly selected and blood was obtained by cardiac puncture for serum thyroxine (T4) and triiodothyronine (T3) concentrations. These pups were then killed and the thyroids were harvested and examined histologically. This protocol was repeated three times at weekly intervals. There were no significant differences in mean T4 and T3 concentrations between treated and control groups at any time during the study. Mean concentration of serum T4 and T3 did increase significantly over the 3 weeks of study. Histologically, no significant differences in thyroid structure were observed between treated and control groups. The numbers of stillborn or weak pups did not increase in the treatment group. No differences were observed in the length of gestation between the treated and control groups. Administration of sulfadiazine and trimethoprim does not affect the thyroid gland in the neonate. PMID- 8229967 TI - Hormonal background of canine and feline mammary tumours. AB - Ovarian steroid hormones and their synthetic derivatives may enhance mammary tumorigenesis in dogs and cats. In toxicity studies of synthetic progestagens a dose-related effect has been observed in the dog, with low-dose exposure sometimes being protective against mammary tumour development. There is some evidence that steroid dependence, as reflected by the presence of steroid receptors (that are nearly always present in normal mammary tissue and benign mammary tumours), is decreased in advanced stages of malignant disease, both in the dog and cat. However, this difference in steroid receptor expression between benign and malignant conditions is not related to any significant alterations in the concentration of receptors for epidermal growth factor. Progestagens have been suggested to promote mammary tumorigenesis in the dog by their induction of growth hormone overproduction; however, there is no conclusive evidence that this effect is necessary for mammary tumour induction. Basal levels of growth hormone and of prolactin were found to be similar in tumour-bearing dogs and age-matched controls. PMID- 8229968 TI - Canine mammary tumour cell lines established in vitro. AB - Mammary tumours are the most common tumours in the female dog. The tumours have a complex histology and exist in epithelial, mixed and mesenchymal forms. To study the biology of canine mammary tumours, five cell lines have been established and characterized. The results indicate that canine mammary tumours might be derived from mammary stem cells and that the tumour growth is independent of oestrogens. The established canine mammary tumour cell lines will be valuable tools in further studies of the histogenesis and pathogenesis of these tumours. PMID- 8229969 TI - Receptors for oestrogen, progesterone and epidermal growth factor in normal and tumorous canine mammary tissues. AB - Receptors for oestrogen (ER), progesterone (PR) and epidermal growth factor (EGF R) are found in normal mammary tissue (NMT) and/or mammary tumours (MT) from all species studied, including dogs. The aims of this study were to define the possible influences of mammary histology, age, location in the mammary chain and of hormonal status and cycle stage in the expression of ER, PR and EGF-R in mammary tissues from healthy dogs and from dogs with MT. Carcinomas that had lost their glandular structure had significantly lower amounts of receptors. NMT either from healthy or affected dogs had significantly higher amounts of ER than MT. PR levels were significantly higher in benign lesions than in NMT. Steroid receptors in NMT from healthy dogs varied significantly with age (older dogs having more ER), location (posterior glands having higher ER concentrations) and cycle stage (the highest ER concentrations being found in the mid-luteal phase and the lowest PR concentrations in the early luteal phase). In NMT from affected dogs, higher steroid receptor concentrations were found in posterior glands; as in healthy dogs, ER concentrations were low in the follicular phase and high in the luteal phase and PR were high in anoestrus. Steroid receptor content in MT did not vary significantly with age, location or cycle stage or with hormonal status, but tended to vary with cycle stage in a manner similar to that found in NMT from the same dogs. In dogs affected with MT and treated with medroxyprogesterone acetate (MPA), NMT had low concentrations of PR but MT from the same dogs had high PR concentrations. EGF-R were found in the majority of the samples (+/- 65% of MT and +/- 85% of NMT) but there was no significant relation between the concentrations and the parameters studied. Nevertheless, EGF-R content was higher in NMT in the proliferative stages (oestrus, early and mid luteal phase) than in the non-proliferative stages (early pro-oestrus and anoestrus). EGF-R and ER were significantly and positively correlated only in malignant tumours. There is no apparent difference between affected and healthy dogs in the regulation of ER, PR and EGF-R expression in NMT; on the other hand, some differences between NMT and MT are observed in the regulation of PR (for example under the influence of MPA) and in the correlation between EGF-R and ER expression. PMID- 8229970 TI - Oestrus induction in the bitch with the synthetic oestrogen diethylstilboestrol. PMID- 8229971 TI - Determination of ovulation in the bitch with a qualitative progesterone enzymeimmunoassay in serum, plasma and whole blood. PMID- 8229972 TI - Effects of contraceptive doses of the progestagen megestrol acetate on luteinizing hormone and follicle-stimulating hormone secretion in female dogs. PMID- 8229973 TI - Synchronous delayed oestrus in beagle bitches given infusions of gonadotrophin releasing hormone superagonist following withdrawal of progesterone implants. PMID- 8229974 TI - Use of a progesterone enzymeimmunoassay to determine the optimal mating time in oestrous bitches. PMID- 8229975 TI - Endoscopic examination of the reproductive tract of the bitch. PMID- 8229976 TI - Study of an enzyme-linked immunosorbent assay for progesterone concentrations in dog plasma. PMID- 8229977 TI - Studies on freezing dog spermatozoa: effect of glycerol on motility after thawing. PMID- 8229978 TI - Studies of protein synthesis in dog oocytes and early embryos. PMID- 8229979 TI - Use of a rapid, qualitative enzyme-linked immunosorbent assay to measure serum progesterone concentration in cats. PMID- 8229980 TI - Gonadotrophin-releasing hormone challenge to test for the presence of ovaries in the bitch. PMID- 8229981 TI - Hysterography as a diagnostic aid in the bitch. PMID- 8229982 TI - Gestagens in the faeces of mink and cats for monitoring corpus luteum activity. PMID- 8229983 TI - Investigation of the endocrine control of parturition in the dog by application of an antigestagen. PMID- 8229984 TI - Recent investigations of the efficacy and safety of proligestone for the prevention of oestrus and pseudopregnancy in bitches. PMID- 8229985 TI - Ovulation without cervical stimulation in domestic cats. AB - Progesterone was measured by radioimmunoassay of serum collected at monthly intervals for 9 months (April-December) in 24 adult female American short-hair cats (age 2.5-11 years, mean 7.4 years); 20 cats were intact, and four were ovariohysterectomized controls. One of the 20 intact queens was ovariohysterectomized after 7 months, when pyometra was diagnosed. Cats could see and hear one another, and could see and hear male cats housed individually in the same room. Direct contact with other cats was prevented. Tactile stimulation of the cats' hindquarters and perineal regions by handlers was avoided. Serum progesterone concentration > or = 4.8 nmol l-1 was defined as evidence of ovulation. This concentration was exceeded in seven of 20 intact queens (35%) at one or more occurrences of non-coital ovulation; there were 13 such occurrences in all (1-3 per queen). Serum progesterone concentration ranged from 0.2 to 103.4 (mean 14.09 +/- 2.0) nmol l-1 in these seven cats, and was significantly greater than concentrations in the other intact and neutered cats. In the remaining 13 intact and four ovariohysterectomized cats, serum progesterone concentrations ranged from 0.2 to 3.2 (mean 1.24 +/- 1.46) nmol l-1. These data suggest that, at least as far as cats housed in proximity to one another are concerned, intact female cats do not always require cervical stimulation to induce ovulation. PMID- 8229986 TI - Fertilization time in the bitch in relation to plasma concentration of oestradiol, progesterone and luteinizing hormone and vaginal smears. AB - Seven bitches of several breeds were monitored during oestrus by vaginal smears and luteinizing hormone (LH), oestradiol and progesterone plasma assays. Each bitch was inseminated into the uterus with frozen spermatozoa from seven dogs of different breeds. Each female was inseminated daily with about 200 x 10(6) motile spermatozoa. Six bitches delivered. The paternity of puppies (which indicated the fertilization time) was determined by morphological analysis. Two bitches were fertilized with one artificial insemination (AI) only and four bitches with two successive AIs. Fertilization occurred 84-180 h after the LH peak and 1-3 days before the first day of metoestrus. The gestation length, from fertilization to birth, was 58-63 days or 56-60 days after the first day of metoestrus. PMID- 8229987 TI - Endocrinology of anoestrous bitches. AB - Plasma luteinizing hormone (LH), prolactin and oestradiol concentrations and responses to a standard challenge with a gonadotrophin-releasing hormone (GnRH) analogue were measured at defined stages of consecutive oestrous cycles in beagle bitches. In each of 35 sampling sequences blood samples were collected every 20 min for 6 h followed immediately by injection of a GnRH analogue (GnRH-A) and collection of further samples after 10, 20, 40 and 60 min. Cycle stages were based on progesterone profiles and the day of the preovulatory LH peak. Sampling sequences were obtained during the luteal phase (n = 5), transition to anoestrus (n = 10), anoestrus (n = 17) and pro-oestrus (n = 3), which were 154-71, 114-44, 85-11 and 7-1 days before the preovulatory LH peak, respectively. Pulsatile LH secretion occurred spontaneously at all stages of the cycle and there was no effect of cycle stage on mean LH concentration or variability. In contrast, oestradiol was undetectable in most samples from early and mid-anoestrus until approximately 1 month before the preovulatory LH peak, after which average oestradiol concentrations and between-sample variability appeared to increase. Mean (+/- SEM) oestradiol concentrations for all preinjection samples collected from 100-75, 74-50, 49-25, 24-10 and 9-1 days before the LH peak was 1.4 +/- 0.1, 1.3 +/- 0.1, 2.4 +/- 0.3, 11.0 +/- 1.4 and 36.0 +/- 3.2 pg ml-1, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229988 TI - Concentrations and pulse analyses of adrenocorticotrophin and luteinizing hormone in plasma from dogs in pro-oestrus, oestrus, dioestrus or anoestrus. AB - Concentrations of adrenocorticotrophin (ACTH) and luteinizing hormone (LH) were measured in plasma collected every 5 min for 190 min from bitches during the oestrous cycle to determine whether mean concentration of ACTH in plasma changes with oestrous cycle phase, and to determine whether plasma ACTH peaks, if present, are related to LH peaks. Radioimmunoassays (RIA) validated for use with dog plasma were used. Mean concentration of ACTH in plasma from pro-oestrous dogs [12.7 +/- 6.0 (SD) range: 5.7-23.1 pmol l-1] was slightly higher (P = 0.085) than that in dogs in oestrus (7.9 +/- 2.1, range: 5.0-11.9 pmol l-1), dioestrus (7.8 +/- 1.5, range: 6.0-9.8 pmol l-1), or anoestrus (7.5 +/- 1.5, range: 5.2-9.2 pmol l-1). Mean concentration of LH in plasma from pro-oestrous dogs [6.0 +/- 7.6 (SD), range: 1.0-19.1 ng ml-1] was higher (P < 0.05) than that in dogs in oestrus (0.6 +/- 0.8, range: undetectable to 2.2 ng ml-1), dioestrus (1.6 +/- 2.1, range: 0.7-5.9 ng ml-1), or anoestrus (1.4 +/- 1.3, range 0.1-3.4 ng ml-1). Plasma ACTH and LH peaks were detected in some dogs during each phase of the cycle. Mean frequency of ACTH peaks (range 0-3 peaks) was similar in all oestrous cycle phases. When data from all phases were combined, mean (+/- SD) number of ACTH peaks in 190 min, interpeak interval and peak duration were 1.7 +/- 1.1, 50 +/- 21, and 34 +/- 7.7 min, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229989 TI - Secretion and release of luteinizing hormone during the luteal phase of the oestrous cycle in the dog. AB - In order that the luteotrophic activity of luteinizing hormone (LH) in the dog might be more precisely elucidated, LH and progesterone were measured in blood samples collected at intervals of 10-15 min over 360 min from three beagle and two foxhound-boxer bitches at a defined day during pro-oestrus, the phase immediately after ovulation, early and late dioestrus and early anoestrus. In addition, the three beagle bitches were treated with the gonadotrophin-releasing hormone (GnRH) analogue buserelin at a given day during each quarter of dioestrus and blood was sampled over 480 min at intervals of 15 min (for 2 h), 30 min (for 4 h) and 60 min (for 2 h). Cyclic activity was monitored by determining progesterone at intervals of 1 or 2 days up to 140 days. In spite of some apparently breed-specific differences, the results indicated that the availability of LH depended on the stage of the cycle. The area under the curve (AUC) was higher during pro-oestrus, late dioestrus and early anoestrus (P < 0.01 0.05), largely as a result of an increased pulse amplitude and not of changes in basal concentrations or pulse frequency. After GnRH the AUC for the provoked LH release increased from 1206 ng (150 min) ml-1 (first quarter) to 4045 ng (150 min) ml-1 (fourth quarter) (P < 0.01); maximum LH values were reached between 25 and 40 min. In neither experiment did progesterone increase during the observation periods of 360 and 480 min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229990 TI - Ultrasonographic monitoring of follicular development, ovulation and the early luteal phase in the bitch. AB - Follicular development, ovulation and the early luteal phase in 15 oestrous cycles of 13 individual bitches were monitored daily by ultrasonography using a 7.5 MHz sector scanner transducer. These findings were compared with vaginal cytology and changes in the peripheral plasma concentrations of oestradiol, luteinizing hormone (LH) and progesterone. Both ovaries were imaged throughout the period of study. Follicles were identified as anechoic spherical structures as early as day 1 of pro-oestrus in two cases and on days 3-4 in 13 cases. The average follicular size increased from 3.7 +/- 0.6 mm to 6.9 +/- 0.7 mm (P < 0.001) between day -5 and the LH surge (day 0). The anechoic structures then increased in size to 7.5 +/- 0.7 mm (P < 0.05) until day 2 after the LH surge; their diameters continued to increase during corpus luteum development, up to 8.1 +/- 0.6 mm (P < 0.001) on the first day of dioestrus. The average number of follicles counted imaged on the day of the LH surge was 3.3 +/- 0.4 on the left ovary and 3.7 +/- 0.7 on the right ovary. The rapid disappearance of the anechoic antrum, corresponding to ovulation, was detected in only two bitches. In the remaining cases follicular rupture was not detected; however, starting at the LH surge, a gradual thickening of the antral wall occurred. This corresponded to a progressive obliteration of the anechoic region and was characteristic of the postovulatory corpora lutea. These changes led to a significant decrease (P < 0.05) in the number of anechoic structures to 2.4 +/- 0.5 on the left ovary and 3.3 +/- 0.5 on the right ovary by day 1 of dioestrus. During the follicular phase, the mean increase in oestradiol concentration was correlated with follicular size (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8229991 TI - Induction of embryonic major histocompatibility complex antigen expression by gamma-IFN. AB - Preimplantation mouse embryos were incubated in vitro with mouse recombinant gamma-interferon (IFN). The effect of the gamma-IFN on major histocompatibility complex (MHC) class I antigen expression was tested using an ELISA procedure. It was found that there is a doubling of Db antigens and a tripling of Qa-2 antigens on C57BL/6 mouse embryos cultured from the 8-cell stage for 24 h in the presence of 10(5) units/ml gamma-IFN. The effect of gamma-IFN on the rate of preimplantation embryonic development was tested by culturing 2-cell embryos for 48 h and 8-cell embryos for 24 h in the presence of varying concentrations of gamma-IFN up to 10(6) units/ml. Two methods were used to assess the cell number per embryo after the culture period: incorporation of [3H]thymidine into DNA, and direct counting of nuclei in fixed and stained embryos. Both methods showed that treatment with gamma-IFN increases the rate of development of preimplantation mouse embryos. Since rate of preimplantation embryonic development is genetically controlled by the Ped gene, it is suggested that gamma-IFN has a direct effect on the Ped gene phenotype of preimplantation mouse embryos. PMID- 8229992 TI - Regulation of the macrophage population in postnatal rat testis. AB - Testicular macrophages increase in concentration during postnatal development in rats. This process may be under hormonal control since administration of hCG stimulates a similar increase to occur precociously. The purpose of the present studies was to determine how the macrophage population is regulated during normal postnatal development and in response to exogenous hCG. We first determined that testicular macrophages proliferate in situ during development and that hCG administration results in an increase in proliferation when given to 10-day-old rats. We next evaluated whether hCG might exert its effects through enhanced secretion of testosterone from Leydig cells. We found that testosterone could not induce a precocious increment in the macrophage concentration when it was administered to newborn pups for 10 days. Finally, the normal increase in macrophage concentration that occurs prior to puberty could not be blocked by treatment with the antiandrogen Casodex. The results are consistent with the hypothesis that the macrophage population expands by proliferation, perhaps under gonadotropin control. In addition, neither the precocial expansion that occurs in response to hCG nor the normal expansion that occurs before puberty is mediated by testosterone. PMID- 8229993 TI - Elevated endometrial natural killer cell activity during early porcine pregnancy is conceptus-mediated. AB - This study investigated an extended time course of endometrial NK cell activity during gestation and the mechanisms underlying changes in uterine NK cell activity in pigs. Endometrial tissues were collected from cyclic, pseudopregnant and pregnant nulliparous pigs on various days post-estrus, and from pigs 10 days after insemination with seminal plasma or killed spermatozoa. NK effector cells were isolated from each endometrial sample, size fractionated and tested for cytolytic activity against NK target cells (K562) using chromium release assays and immunocytochemically for the frequency of perforin-positive cells. Various cell fractions showed different levels of NK activity and had different proportions of cells expressing perforin. Morphologically, cells in the fraction with maximal NK activity almost all showed typical lymphocyte size and shape. Substantially elevated NK cell activity was recorded in pregnant pigs on days 10 and 20 of gestation. By day 30, the cytolytic activity declined dramatically to an almost undetectable level. Very little activity was found in uterine cells isolated from cyclic, pseudopregnant, and seminal plasma or killed spermatozoa inseminated animals, and no differences were detected either between follicular and luteal phases of the estrous cycle or between different days of pseudopregnancy. These results indicate that elevated NK cell activity during early porcine pregnancy cannot be attributed to contributions from either the maternal systemic endocrine status or from components of boar semen. The changes in NK cell activity observed in porcine endometrial tissues during early pregnancy must therefore be associated with the actual presence of conceptuses. PMID- 8229994 TI - Mouse metrial gland cells do not kill Yac-1 myeloma cells. AB - Mouse metrial gland cell suspensions, which included granulated metrial gland cells, were assessed for their ability to lyse NK cell target Yac-1 myeloma cells in a 51chromium release cytotoxicity assay. Metrial gland cells did not kill Yac 1 cells even after in vivo stimulation of NK cytotoxicity activity by polyinosilic-cytidilic acid. The precise relationship of granulated metrial gland cells to the NK cell lineage remains to be clarified. PMID- 8229995 TI - Characterization of antibodies induced by paternal lymphocyte immunization in couples with recurrent spontaneous abortion. AB - This study was designed to identify and characterize the allo- and autoantibodies induced following successful paternal lymphocyte immunization to prevent recurrent spontaneous abortion. Firstly the titers of maternal anti-paternal antibodies in women with successful pregnancies as determined by the flow cytometry crossmatch (FCXM) were highly variable; however, in all cases, the initial pre-immunization titers were negative and the post-immunization titers were positive by the FCXM in successfully treated women. Secondly, the specificities of maternal alloantibodies to paternal HLA antigens (immunogen) were evaluated. No all predicted antibodies to mismatched paternal HLA antigens were found by microlymphocytotoxicity (MCX) assays and the specificities varied. Thirdly, antibodies in post- but not preimmunization sera reacted with two lymphoid cell lines, SupT1 and SB; in addition, the rise and fall of the titers of these sera with paternal cells seemed to be reflected with the cell lines by the FCXM. Fourthly, autoantibodies to activated lymphocytes were detected and seemed to correlate with successful immunization since women who had another abortion following immunotherapy lacked these autoantibodies. These findings suggest that the antibody response following successful immunotherapy is complex and needs to be studied further to understand the mechanism of this treatment. PMID- 8229996 TI - Pregnancy-associated, lymphocyte-derived suppressor factor inhibits protein kinase C activity. AB - For successful allogenic pregnancy to occur, suppression of maternal defense responses toward the fetus are vital. Suppressor factors elaborated by decidual cells or immune cells may facilitate this suppression. In order for appropriate cellular responses to occur an intact signal transduction/second messenger system must be present. The calcium/phospholipid-dependent protein kinase, Pk-C, plays an important role in regulating immune responses, and may also be important in regulating uterine cell responses and implantation events. Pk-C activation is necessary for IL-2 synthesis and IL-2 receptor synthesis through activation of the proto-oncogenes c-jun and c-fos. These proto-oncogene gene products combine to form the heterodimer AP-1 which then activates IL-2 gene transcription for both peptide and receptor. If Pk-C activity becomes abrogated then appropriate cell responsiveness is diminished. We have shown that Pk-C activity is decreased in the particulate fraction of 4-7 day pregnant spleen, thymus and draining lymph node (DLN) cells. Spleen cells did not exhibit any change in cytosolic Pk-C activity, the thymus was found to have a decrease in both cytosol and particulate fractions, and the DLN cells exhibited a translocation effect whereby particulate Pk-C decreased and cytosolic Pk-C activity increased. Supernatant from 3-day cultures of DLN cells from pregnant animals was shown to inhibit proliferation of spleen cells. In addition, the supernatant was able to directly lower Pk-C activity. We hypothesize that DLN cells secrete a factor(s) that is able to suppress immune response through abrogation of Pk-C activity, thereby decreasing AP-1 formation resulting in decreased IL-2 synthesis and IL-2 receptor synthesis. PMID- 8229997 TI - Effect of sperm-associated antibodies on human sperm ability to bind to zona pellucida and to penetrate zona-free hamster oocytes. AB - Antibodies directed against sperm membrane antigens located mainly over the sperm heads were eluted from the sperm cell fraction of autoimmune ejaculates and transferred to antibody-negative spermatozoa of fertile donors. The ability of these antibody-coated spermatozoa to bind to the human zona pellucida and to penetrate zona-free hamster oocytes was evaluated in vitro. The majority of the sperm-eluted samples contained both IgA and IgG antibodies. In order to evaluate the effect of each class of antibody on the analyzed sperm functions, each isotype was specifically absorbed before transfer. Sperm-binding to salt-stored zona pellucida, as assayed by FITC and TRITC labeling of antibody-free and antibody-coated spermatozoa incubated with the same zona, was consistently reduced by 60-85% by the five eluted samples tested. Removal of either IgA or IgG antibodies from the eluted samples did not change the overall effect. Sperm penetration of zona-free hamster eggs was variously affected by sperm-associated antibodies. Of the 8 samples of sperm-eluted antibodies tested, only 4 had a significant effect on sperm penetration. Three of them decreased the penetration by 67-78%, while the fourth exhibited a modest increasing effect of 39%. These four samples contained antibodies of the two isotypes. In the samples with a decreasing effect, the elimination of one or another of the two isotypes restored the ability of the sperm to penetrate the hamster oocytes. These results suggest that sperm-associated antibodies may have different effects on zona-binding and gametic fusion events that lead to fertilization. Whereas IgA and IgG antibodies taken together or separately decreased sperm binding to the human zona pellucida, the two classes of antibodies must be associated in order to impair sperm penetration into zona-free hamster oocytes. PMID- 8229998 TI - Reactivities of a monoclonal antibody directed against an immunosuppressive molecule in boar seminal plasma. AB - Using a monoclonal antibody (LIS-4) to an immunosuppressive factor isolated from boar vesicular gland secretion it was determined that this gland secretes a tissue-specific immunosuppressive molecule that is absorbed onto the acrosome of spermatozoa during ejaculation. Absorption of the immunosuppressive molecule onto murine embryos at the 2-, 4-, 8-cell, morula and blastocyst stages in vitro was evaluated by indirect immunofluorescence. In vivo absorption was detected on the zona pellucida of murine embryos obtained from oviducts injected with the immunosuppressive molecule. Immunofluorescence revealed that the immunosuppressive molecule was not absorbed onto murine embryos after solubilization of the zona pellucida. There was no effect of the antibody to the immunosuppressive molecule on the ability of boar spermatozoa to penetrate the porcine zona pellucida. PMID- 8229999 TI - Antibodies to phospholipids and nuclear antigens in non-pregnant women with unexplained spontaneous recurrent abortions. AB - In a collaborative study of 73 non-pregnant Kuwaiti women with unexplained spontaneous recurrent abortion (RSA), 30 control healthy non-pregnant multiparous Kuwaiti women and 20 North American women who received elective abortion(s), autoantibodies to 6 phospholipids and 9 nuclear antigens were measured. Women with recurrent spontaneous abortions demonstrated 3 times higher incidence of antibodies to phospholipids (30.1%) than controls (10% each) (P = 0.029). The incidence of both IgM and IgA class antiphospholipid antibodies were significantly higher than those of controls. The incidence of antibodies to cardiolipin in women with recurrent spontaneous abortions (12.3%) was significantly higher than those of controls (P = 0.035) and incidence of IgM but not IgG anticardiolipin antibody was significantly higher in women with RSAs than in controls (P = 0.053). The incidences of anti-polyinosinic acid (P = 0.035) and anti-histone 1 antibody (P = 0.052) were significantly higher in women with recurrent spontaneous abortions than controls. There was no significant difference in the incidence of autoantibodies between primary and secondary aborters. However, women with a history of second trimester abortions showed a higher incidence of antiphospholipid antibodies than women with first trimester abortions only. Recurrent spontaneous abortion is associated with autoantibodies to phospholipid epitopes including IgA antiphospholipid antibodies. PMID- 8230000 TI - Determination of immunoglobulin classes and subclasses of sperm immobilizing antibodies in infertile women and the inhibitory effect of IgG4 subclass on the sperm immobilizing antibody activity. AB - Immunoglobulin classes and subclasses of sperm immobilizing antibodies (SI-Abs) in the sera of sterile women were determined by the absorption of patients' sera with Staphylococcus aureus and immunoadsorbents bound with class or subclass specific anti-human immunoglobulin antibodies. Among 18 patients' sera tested, 16 had the IgG-dominant SI-Abs and the remaining 2 sera contained IgM-dominant SI Abs. From the former patients' sera, 9 were further studied to determine the IgG subclasses; 6 of them had IgG1-dominant SI-Abs, one IgG2-dominant, and in the other two IgG1 and IgG2 were equally dominant. Interestingly, SI-Ab activities in 6 of the 9 patients' sera increased after absorption of IgG4 subclass and the addition of IgG4 purified from an SI-positive patient to the same patient's serum diminished the SI-Ab activities. PMID- 8230001 TI - Molecular identity of a sperm acrosome antigen recognized by HS-63 monoclonal antibody. AB - The molecular identity of mouse sperm acrosome antigen recognized by HS-63 monoclonal antibody was analyzed by various biochemical, immunological and molecular biological methods. When its cognate antigen, MSA-63 was isolated from mouse testis by immunoaffinity chromatography, a group of protein spots with wide range of molecular sizes and isoelectric points were identified. Through previous studies, it was established that most of these protein spots were actin-like molecules co-purified with MSA-63 protein from mouse testis. To analyze the molecular size heterogeneity of the isolated MSA-63 proteins, rabbit antisera against a computer-predicted antigenic synthetic peptide (amino acid residue No. 160-171) and a recombinant glutathione S-transferase (GST) fusion protein (GST 63) were raised. These two antisera and those raised against the isolated MSA-63 protein were used as the probes in comparative Western blot assay, indirect immunofluorescent assay and enzyme-linked immunosorbent assay (ELISA). Using ELISA, antisera against GST-63 and computer-predicted antigenic synthetic peptides were shown to cross-react with affinity-isolated MSA-63 protein coated on microwells. However, little immunological cross-reactivity was observed between GST-63 fusion protein and the synthetic peptide. By using a Western blot assay, two major protein bands of 22 and 32 kDa, respectively were commonly detected on mouse testis homogenate strips by both anti-MSA-63 and anti-GST-63. In addition, anti-MSA-63 also recognized several protein bands with molecular masses greater than 35 kDa. The results of this study suggested that the molecular heterogeneity of MSA-63 protein isolated from mouse testis and sperm, is due to a series of post-translational modifications on a single gene product. These modifications may include glycosylations, proteolytic digestions and tight non-covalent associations with other testicular cytoskeletal proteins, such as actins. PMID- 8230002 TI - Sperm antibodies in a patient with obstructive azoospermia. AB - An artificial spermatocoele was constructed at the level of the caput epididymis in a patient with obstructive azoospermia. Morphologically normal spermatozoa were harvested but motility and progression were inadequate for intra-uterine insemination or in vitro fertilisation. In addition, there was an antispermatozoal antibody response which transuded back into the reproductive tract and coated spermatozoa with antibody. It is recommended that if harvesting of epididymal spermatozoa is contemplated, the serum should initially be assessed for spermatozoal antibody. PMID- 8230003 TI - Predicting the prognosis of patients with rheumatoid arthritis: is there a crystal ball? PMID- 8230004 TI - Antiphospholipid antibodies and the kidney. PMID- 8230005 TI - Pathogenesis of ankylosing spondylitis: recent advances. PMID- 8230006 TI - Aberrant control of galactosyltransferase in peripheral B lymphocytes and Epstein Barr virus transformed B lymphoblasts from patients with rheumatoid arthritis. AB - It is now well established that hypogalactosylation of IgG is a molecular marker for rheumatoid arthritis (RA). However, the mechanism for the alteration of the galactosylation status has not been resolved. We compared the galactosyltransferase activities of anti-CD19 selected peripheral B lymphocytes of healthy subjects and patients with RA using ovalbumin as the acceptor substrate. In addition, certain samples of lymphocytes were assayed after Epstein Barr virus (EBV) transformation and, also, the ability of bovine milk galactosyltransferase to galactosylate IgG in vitro was examined. Our results indicate that there is a significant difference between the galactosyltransferase activities of rheumatoid and control peripheral B lymphocytes and that EBV transformation causes a variable increase (15-1225%) in galactosyltransferase activity, over that present in the peripheral B lymphocytes from which the transformed cells were derived. Also the ubiquitous "lactose synthetase" type galactosyltransferase (EC 2.4.1.38) will galactosylate normal native IgG at concentrations of 500 mU/ml in vitro. We conclude that there is no evidence from our study for an IgG specific galactosyltransferase and that galactosyltransferase is an enzyme that is aberrantly modulated in peripheral B lymphocytes and EBV transformed B lymphoblasts derived from patients with RA. PMID- 8230007 TI - Factors predicting outcome of rheumatoid arthritis: results of a followup study. AB - OBJECTIVE: To determine prognostic factors in rheumatoid arthritis (RA). METHODS: One hundred thirty-two women with definite RA were followed yearly from an early phase of the disease (symptoms < 5 years) for a mean duration of 6 years. The prognostic value of the first available clinical and laboratory variables and assessments of functional ability was related to several outcome measures (physician's opinion of disease severity, disease activity, radiological abnormalities, functional ability and number of prescribed 2nd-line drugs) by single predictor analysis and by logistic regression. RESULTS: The variables most predictive for one or more of the outcome measures were number of swollen joints, Ritchie score, health assessment questionnaire score, radiographical abnormalities, positive IgM rheumatoid factor (RF), positive IgG-RF, HLA-DR4, and an elevated percentage serum agalactosyl IgG. The accuracy of predicting outcome was calculated from several combinations of these variables, and varied between 70 and 80%. The accuracy based on a combination of the commonly available variables (number of swollen joints, IgM-RF and the erosion score), closely approximated the maximal accuracy that could be achieved. CONCLUSION: The outcome of RA can be predicted by a combination of variables that are commonly available in the clinical setting. PMID- 8230008 TI - Aspirin is not associated with more toxicity than other nonsteroidal antiinflammatory drugs in patients with rheumatoid arthritis treated with methotrexate. AB - OBJECTIVE: To compare the clinical and laboratory toxicities of aspirin vs nonsteroidal antiinflammatory drugs (NSAID) in combination with low dose methotrexate (MTX). METHODS: We retrospectively examined 34 patients with rheumatoid arthritis (RA) who completed 12 months of a prospective MTX trial. Analysis included descriptive and logistic regression. RESULTS: Twelve patients took an average of 4.5 g aspirin daily; 22 patients took other NSAID at stable doses. Limiting toxicity was not different between aspirin and NSAID treatment groups, respectively, for stomatitis (33 vs 27%), gastrointestinal symptoms (25 vs 18%), hepatic (25 vs 27%), or other toxicity. However, using logistic regression procedures, weight adjusted weekly MTX dose and prednisone dose correlated with toxicity. CONCLUSION: While toxicity is common when aspirin or NSAID are used with MTX to treat RA, there is no clinical difference between aspirin and NSAID with respect to that toxicity during 12 months of therapy. PMID- 8230009 TI - Increased type I collagen gene expression in L-tryptophan associated eosinophilia myalgia syndrome skin fibroblasts. AB - OBJECTIVE: To investigate the pathogenetic mechanisms responsible for the cutaneous fibrosis in eosinophilia-myalgia syndrome (EMS) associated with L tryptophan ingestion. METHODS: Compare in vitro type I collagen production and steady state procollagen mRNA levels in cultured skin fibroblasts derived from healthy individuals and from 5 patients with EMS and diffuse cutaneous induration. RESULTS: Cell lines derived from the affected skin from patients with EMS exhibited greater collagen production and higher steady state levels of alpha 1(I) procollagen mRNA compared with fibroblasts from age and sex matched healthy individuals. Exposure to interferon gamma reduced collagen synthesis in the EMS fibroblast lines. The rate of in vitro transcription of the COL1A1 gene was 30% higher in nuclei isolated from collagen overproducer EMS fibroblasts than in nuclei from normal fibroblasts. CONCLUSION: Fibroblasts derived from the involved skin of patients with EMS show increased expression of the alpha 1(I) procollagen gene in vitro compared to normal skin fibroblasts. The biosynthetically activated phenotype exhibited by EMS fibroblasts appears to be due, at least in part, to transcriptional activation of type I collagen gene expression. These biochemical and molecular alterations may result in accumulation of collagen and lead to the cutaneous fibrosis in EMS. PMID- 8230010 TI - Serum cytidine deaminase as a measure of disease activity in rheumatoid arthritis and systemic lupus erythematosus. AB - Serum cytidine deaminase (CD) as a marker of disease activity was assessed in 100 patients with rheumatoid arthritis (RA) and in 102 assessments of 85 patients with systemic lupus erythematosus (SLE). In RA CD levels correlated well with clinical assessment of disease activity, but were not influenced by varying dosages of ibuprofen as therapy. In SLE significant correlations were found between CD and anti-DNA antibody titers, as well as C3 complement levels. A subset of clinically active patients with SLE with elevated CD levels but normal anti-DNA titers was identified. Serum CD levels may be a clinically useful marker in RA and in certain subgroups of patients with SLE. PMID- 8230011 TI - Systemic lupus erythematosus and Hodgkin's lymphoma. AB - OBJECTIVE: Systemic lupus erythematosus (SLE) is associated with lymphoproliferative diseases such as Hodgkin's lymphoma. Since there is considerable overlap between the features of SLE and Hodgkin's lymphoma (SLE/HL) there can be a great difficulty in diagnosing Hodgkin's lymphoma in the presence of SLE. Our objective was to determine the features that can suggest coexistent SLE/HL: METHODS: We describe 3 cases of Hodgkin's lymphoma that occurred in the setting of SLE and review the features of 11 reported cases in which the diseases occurred simultaneously. We compare the features of these cases with those of Hodgkin's lymphoma and SLE occurring alone. RESULTS: Mediastinal adenopathy was seen in 58% and retroperitoneal adenopathy in 17% of patients with SLE/HL, while it was seen only rarely in these sites in cases with SLE. Severe generalized pruritus, which occurs in only 2.8% of cases of SLE, was seen in 63% of cases with SLE/HL: Features such as fever, weight loss, peripheral lymphadenopathy, splenomegaly and hepatomegaly are common to both Hodgkin's lymphoma and SLE. Renal disease is seen in half of the cases of SLE. It is rare in Hodgkin's lymphoma where, unlike SLE, it is only of minimal change or membranous type. Arthritis and positive antinuclear antibodies are rare in Hodgkin's lymphoma. CONCLUSIONS: Persistent lymphadenopathy, especially mediastinal and/or retroperitoneal lymphadenopathy, eosinophilia and generalized pruritus in a patient with SLE not responding to treatment may be indicative of coexistent Hodgkin's lymphoma. PMID- 8230012 TI - Anticardiolipin antibodies and dependence of a serum cofactor. A mechanism of thrombosis. AB - OBJECTIVE: To evaluate the dependence on the serum cofactor of anticardiolipin antibodies (aCL) in infectious and autoimmune diseases. We also studied their correlation with some clinical manifestations, specially thrombosis. METHODS: aCL were determined with a standard ELISA method, and a modified ELISA in which we substituted bovine serum albumin (BSA), gelatin and skim milk powder for fetal calf serum (FCS). Categorized variables were analyzed by means of the chi 2 test and Fisher's test. Four groups of patients were studied. Group 1. Patients with aCL and autoimmune disease (systemic lupus erythematosus [SLE] and the primary antiphospholipid syndrome [PAPS]). Group 2. Patients with aCL, no symptoms and no underlying infection or autoimmune disease. Group 3. Patients with aCL and infectious diseases (syphilis, leprosy, HIV infection and Q fever). Group 4. Control group. RESULTS: (a) 19 of 20 samples from patients in Group 1 disclosed cofactor dependence in aCL activity. (b) 17 of 19 samples from patients in Group 3 had aCL activity, that was independent of the presence of the cofactor. (c) 3 of 4 patients in Group 2 had cofactor independent aCL and one had cofactor dependent aCL activity. (d) no control group patient had aCL. (e) association of cofactor dependent aCL with the development of clinical manifestations (thrombosis) was statistically significant (p < 0.0001). (g) cofactor dependent aCL and cofactor independent aCL were, respectively, associated with autoimmune and infectious diseases (p < 0.0001). CONCLUSIONS: (a) Dependence or independence of the cofactor helps to differentiate "infectious" aCL from "autoimmune" aCL. (2) aCL related clinical manifestations (thrombosis) depends on the presence of cofactor dependent aCL and not on cofactor independent aCL. PMID- 8230013 TI - Mechanisms of endothelial cell damage in systemic sclerosis and Raynaud's phenomenon. AB - OBJECTIVE: To investigate tumor necrosis factor (TNF), circulating immune complexes (CIC) and oxidized lipoproteins [measured as thiobarbituric acid reactive substances (TBARS)] as mediators of damage to endothelial cells (EC) in systemic sclerosis (SSc) and Raynaud's phenomenon (RP). METHODS: In addition to CIC, TBARS and TNF, von Willebrand factor (vWF), angiotensin converting enzyme (ACE) and EC cytotoxicity were measured as markers of EC damage. C-reactive protein (CRP) and rheumatoid factor (RF) were measured as indicators of inflammation. RESULTS: There were increases in TBARS and CIC, vWF and EC cytotoxicity, and CRP, with reduced levels of ACE. There were no correlations between any possible mediator of damage (TNF, TBARS, CIC) and indicators of such injury to the endothelium (vWF, ACE, EC cytotoxicity) except an inverse correlation between vWF and ACE. When patients with SSc were classified into those with limited disease (ISSc) or diffuse disease (dSSc), only wWF was different, being higher in patients with dSSc. In RP, vWF, RF and TBARS were again raised and ACE was lower. The only correlation was inversely between vWF and ACE. CONCLUSION: Our results are suggestive of injury to the endothelium in both SSc and RP, with greater damage in patients with dSSc. Although there were multiple abnormalities in biochemical and immunological indices, there were no consistent correlations to explain the clear damage to the vasculature. We conclude that additional mechanisms other than TNF, CIC and lipid peroxides may be responsible for the insult to the vascular tree so evident in these patients. PMID- 8230014 TI - Sensitive rapid detection of myoglobin in serum of patients with myopathy by immunoturbidimetric assay. AB - OBJECTIVE: To study immunoturbidimetry, a new technique for the quantitative determination of myoglobin in serum. METHODS: Correlation with the standard technique of radioimmunoassay in sera of patients with and without myopathy. RESULTS: Immunoturbidimetry was found to have a close correlation with the radioimmunoassay (RIA) method (r = 0.97, p = < 0.001), although it was slightly less sensitive. Compared to RIA, sensitivity was 87.8% and specificity, 98.6%. The turbidimetric method was performed considerably more rapidly than RIA. Using this method to assess patients with dermatomyositis and polymyositis, a close relationship to clinical findings was noted. However, of 64 samples with myoglobin level greater than 50 ng/ml, there were 3 instances (4.7%) where elevated myoglobin in the presence of normal CK suggested active disease, and there were 9 instances (14.1%) where myoglobin was normal in the presence of elevated CK. Seven of these cases were felt to reflect a lower sensitivity of the turbidimetric method. The other 2 are not explained. Serial measurements of serum myoglobin correlated well with clinical state in the course of 2 patients illustrated. CONCLUSIONS: Assay of serum myoglobin by immunoturbidimetry may provide a rapid and easily performed method without reliance on radiochemicals. It can be useful in clinical assessment of patients with myopathies, although sensitivity is not quite as high as that of RIA. PMID- 8230015 TI - Cancer in association with polymyalgia rheumatica and temporal arteritis. AB - In our prospective controlled study, a total of 185 patients with polymyalgia rheumatica (PMR) and temporal arteritis (TA) diagnosed during 1978-83 and their 925 matched controls were cross checked with the data files at the Cancer Registry of Norway at the end of 1987. Malignancy was registered in 27 patients (14.6%) and 131 controls (14.2%) between 1953 and the end of 1987. Malignancy was registered in 16 (24.6%) of the patients with biopsy demonstrating arteritis temporalis. The hazard rate for developing malignancy after diagnosis for the whole patient population was not significantly different from the controls. The hazard rate for developing malignancy in patients with positive biopsy, however, was 2.35 times higher than in the controls (p = 0.036) and 4.40 times higher than the rest of the patient population (p = 0.007) (Cox proportional hazards model). The general long interval between diagnosis of PMR and/or TA and registration of malignancy (mean 6.5 years) is not consistent with a paraneoplastic mechanism. PMID- 8230016 TI - Resistive exercise in the rehabilitation of polymyositis/dermatomyositis. AB - OBJECTIVE: To determine the effect of a resistive exercise program in the rehabilitation of patients with polymyositis/dermatomyositis (PM/DM). METHODS: Five patients participated in a rehabilitation program that included resistive exercises. In 4 of the patients, these exercises were randomly alternated with nonresistive exercise. Each type of exercise was assigned for periods of 2 weeks, and at the end of each one, muscle strength was measured by means of manual muscle testing (MMT), an activities of daily living (ADL) score, and peak isometric torque (PIT) generated by muscle groups in the lower extremities. Creatine phosphokinase (CPK) levels were used to measure disease activity. The acute response to resistive exercise was studied in 4 of the patients by measuring serial levels of CPK following a session on an exercise bicycle. RESULTS: Of the 4 patients who participated in both types of exercises, 3 experienced increases in strength during both resistive and nonresistive exercise periods. The patient who participated only in resistive exercises had considerable improvements in muscle strength. One patient made no improvements in strength with either type of exercise. None of the patients experienced clinically significant elevations in CPK attributable to either type of exercise. Following a session of resistive exercise, the mean CPK elevation in 4 patients was 7.7%, returning to pre-exercise levels by 8 h in all cases. CONCLUSIONS: Patients with PM/DM may participate in a rehabilitation program which includes resistive exercises. Such programs may be accompanied by increased strength without clinically significant rises in serum levels of muscle enzymes. PMID- 8230017 TI - Host risk factors for the development of reactive arthritis: a family study. AB - OBJECTIVE: To determine if the same arthritogenic microorganisms can trigger reactive arthritis (ReA) in both HLA-B27 positive and HLA-B27 negative individuals. To determine risk factors (other than HLA-B27) for the development of ReA. METHODS: Following a common food source outbreak of Salmonella enteritidis, 2 family groups of individuals with dysentery and new onset ReA signs and symptoms were identified. HLA typing was performed for all family members exposed to the dysenteric organism. RESULTS: ReA occurred in HLA-B27 positive members of Family A. No member of Family B possessed HLA-B27 or CREG phenotype, but 3 members developed ReA. The HLA identical sibling of the proband in Family B developed dysentery but did not develop ReA. CONCLUSION: In this epidemic, interaction of the microorganism with an HLA-B27 positive host predictably produced ReA but B27 was not necessary to produce it in other dysenteric patients. Host factors including amount of ingested microorganisms, age and sex but excluding HLA type may account for the differential development of ReA in the siblings in Family B. PMID- 8230018 TI - A prospective study of patients with spondyloarthropathy with special reference to HLA-B27 and to gut histology. AB - Ileocolonoscopy was performed on 357 patients meeting the European Spondylarthropathy Study Group criteria for spondyloarthropathy. HLA loci A, B and C were determined in all patients; HLA-B27 was detected in 196 and was absent in the 161 remaining patients. A number of clinical, laboratory and radiological variables were determined before ileocolonoscopy and compared between the HLA B27+ and HLA-B27- patients. The HLA-B27+ patients were mainly men, with significantly more family members with spondyloarthropathies. Clinical evidence of tendinitis and uveitis was more frequently found in these patients. Like several authors, we found that these patients were more severely affected since they presented more severe radiological involvement of the sacroiliac joints. Syndesmophytes, bamboo spine and erosive joint lesions were more frequent in this group, the hip involvement being of the concentric type. In HLA-B27+ patients ankylosing spondylitis was more prevalent, while in the B27- patients enterogenic, urogenital or undifferentiated spondyloarthropathy was diagnosed. The HLA-B27- patients experienced more episodes of diarrhea, and Crohn-like inflammatory gut lesions were more frequently seen on ileocolonoscopy. In this group a number of patients probably had a form of subclinical Crohn's disease of which the locomotor symptoms were the only clinical expression. PMID- 8230019 TI - Shoulder joint movement and its relationship to disability in the elderly. AB - One hundred healthy subjects over the age of 65 years were interviewed and examined. The range of movement of their shoulder joints was recorded. Assessment of the prevalence of symptomatic shoulder joint disorders and their nature was also made. The impact of the presence and severity of such disorders on their functional independence was evaluated by means of the Katz Scale of Index of Independence in the Activities of Daily Living. Ranges of all movements were considerably reduced in this age group compared to figures for a younger population. The prevalence of symptomatic shoulder disorders was 34% of which a rotator cuff lesion was the most common. Disability related to these disorders was found in 30% of the study population and was significantly associated with reduced movement. It is suggested that treatment of these disorders might improve quality of life and that routine examination of the shoulder in the aging population would identify those at risk of losing their independence. PMID- 8230020 TI - Stromelysin, tissue inhibitor of metalloproteinases and proteoglycan fragments in human knee joint fluid after injury. AB - OBJECTIVE: To determine in a cross sectional study the concentrations of stromelysin, tissue inhibitor of metalloproteinases (TIMP), and proteoglycan fragments in knee synovial fluid (SF) at different times after injury to cruciate ligament or meniscus. METHODS: Joint fluid samples were obtained from patients with knee injury diagnosed by arthroscopy. Concentrations of stromelysin-1 and TIMP-1 were determined by immunoassay with monoclonal and polyclonal antibodies. Cartilage proteoglycan fragments were quantified by immunoassay with polyclonal antibodies or by dye precipitation. RESULTS: Average concentrations of stromelysin increased 40-fold in association with injury, and after about 6 months decreased to a plateau level about 10-fold increased compared to a reference group with healthy knees. TIMP and proteoglycan levels also increased in similar temporal patterns, but less markedly. Increased average SF levels of these markers were maintained for at least 17 years after injury. SF from knees with injury contained a 1.5 to 2.5 molar excess of stromelysin over TIMP, while reference joint fluids contained a 2-fold molar excess of TIMP over stromelysin. CONCLUSION: The persistent changes in SF markers after joint injury may be associated with the cartilage destruction and frequent development of posttraumatic osteoarthritis in this group of patients. PMID- 8230021 TI - Bone mineral density in patients with temporal arteritis and polymyalgia rheumatica. AB - OBJECTIVE: To assess the influence of corticosteroids in the mineral content of patients with giant cell arteritis (GCA), 56 patients, 28 with polymyalgia rheumatica (PMR) and 28 with temporal arteritis (TA) were studied. METHODS: The bone mineral density (BMD) in the lumbar spine and the femoral neck was measured by dual photon X-ray absorptiometry. A control group (48 people) comparable in age and sex was also evaluated. RESULTS: Compared with the controls, the patients with GCA had lower values of BMD in the femoral neck (men: p < 0.03; women: p < 0.001). In the lumbar spine differences were significant in women (p < 0.05) but not in men (p < 0.1). Multiple regression analysis showed that the BMD at L2-L4 in men correlated with height and weight and was inversely related to the cumulative dose of corticosteroids and the duration of treatment. The BMD at L2 L4 in women correlated with height and weight, but not with corticosteroids. Multiple regression analysis in men showed that age and the total dose of corticosteroids were significant independent predictors of femoral BMD. In women BMD in the femoral neck was correlated with age and weight, but not with corticosteroid treatment. CONCLUSION: The total dose and duration of corticosteroid treatment have been shown to be determinant factors of bone mass in patients with GCA who received corticosteroids. PMID- 8230022 TI - Platelet activating factor and histamine effects in synovial blood flow evaluated by laser-Doppler flowmetry. AB - OBJECTIVE: To investigate platelet activating factor (PAF) and histamine effects in synovial circulation of rabbits. METHODS: A laser-Doppler flowmeter was adapted to study the synovial blood flow. Either PAF or histamine were infused in the first femoral artery branch. The effects of PAF and histamine antagonists as well as of inhibitors of cyclooxygenase products were evaluated. RESULTS: PAF induced a vasoconstriction in synovial circulation, which was inhibited by the PAF antagonist, WEB 2170, a cyclooxygenase inhibitor, indomethacin or a thromboxane synthesis inhibitor, dazmegrel. Histamine induced vasoconstriction in the synovial vessels. Promethazine not only inhibited this vasoconstriction but also induced a vasodilation, that was blocked by a combined treatment with promethazine and cimetidine. Pretreatment with WEB 2170 did not interfere with histamine effect. CONCLUSIONS: Our data indicate that PAF induces vasoconstriction in synovial blood flow through a receptor mediated mechanism and that thromboxane is involved in this effect. Histamine induces constriction or dilation in synovial vessels, through action on H1 or H2 receptors, respectively. This effect is not dependent on PAF. PMID- 8230023 TI - Intraarticular injection of fibronectin fragments causes severe depletion of cartilage proteoglycans in vivo. AB - We have reported that fibronectin fragments (Fn-f) cause cartilage damage in vitro by causing enhanced release of proteases. In order to determine whether the Fn-f can damage cartilage in vivo, we have injected native fibronectin (Fn) and Fn-f into adolescent rabbit knee joints. After 7 days, tissue was analyzed by histochemical and biochemical techniques and remaining proteoglycans quantified. Injection of 0.6 or 3 microM Fn-f caused up to a 70% loss in total cartilage proteoglycan while native Fn, rabbit serum albumin or an Arg-Gly-Asp-Ser synthetic peptide, derived from the cell-binding domain of Fn, did not cause damage. Our results suggest that this Fn-f/damage model may be useful for generating cartilage damage in vivo for other studies. Since Fn-f have been detected in synovial fluids from joints of patients with rheumatoid arthritis and osteoarthritis, our results are consistent with the notion that Fn-F mediated damage may occur in vivo. PMID- 8230024 TI - Withdrawal of longterm antihyperuricemic therapy in tophaceous gout. AB - OBJECTIVE: To determine the effect of withdrawal of antihyperuricemic therapy on the recurrence of tophi and acute arthritis. METHODS: We performed a retrospective investigation of 21 patients with tophaceous gout who stopped therapy because they had no arthritis for years, and on examination tophi had disappeared. RESULTS: Nine patients (43%) had a recurrence of tophi after an average period of 39.6 months (range 4-107 months). Seventeen patients (81%) had attacks of acute arthritis, 6 patients within 6 months. During a mean period of drug discontinuation of 37.6 months no increase in serum creatinine or deterioration in joint function was observed. CONCLUSION: Based on our data we conclude that intermittent usage of antihyperuricemic drugs will, after a variable period, result in the frequent recurrence of acute arthritis and tophi. PMID- 8230025 TI - Cyclosporine A induced colitis and acquired selective IgA deficiency in a patient with juvenile chronic arthritis. AB - A 12-year-old girl with juvenile chronic arthritis was treated with cyclosporine A for recurrent uveitis. Two years later she presented with weight loss and diarrhea. Investigations suggested drug induced colitis. She improved after withdrawal of cyclosporine and her nonsteroidal antiinflammatory drug. Symptoms recurred on rechallenge with cyclosporine. She has developed a progressive selective IgA deficiency which persists despite cessation of cyclosporine. PMID- 8230026 TI - Isometric exercise increases strength and does not produce sustained creatinine phosphokinase increases in a patient with polymyositis. AB - A patient with active stable idiopathic polymyositis received a 4-week supervised right quadriceps and biceps isometric strengthening program. He demonstrated a significant increase in isometric peak torque without a sustained rise in creatinine phosphokinase (CPK). A significant decrease in postexercise CPK occurred at the end of the exercise program. We conclude a randomized clinical trial using a nonexercised patient group vs an exercised patient group may be useful. PMID- 8230027 TI - Rheumatoid arthritis and vertebral artery occlusion: a case report with angiographic and magnetic resonance demonstration. AB - A 40-year-old woman with rheumatoid arthritis (RA) who presented with vertigo and nausea, was found to have left vertebral artery occlusion on angiography. There was minimal atlantoaxial subluxation and no odontoid herniation. Magnetic resonance imaging (MRI) was suggestive of a left vertebral artery dissection. This is the first report of MRI of vertebral artery occlusion in RA. It will be important to accumulate MR images of this complication in future cases. PMID- 8230028 TI - Adult respiratory distress syndrome associated with antiphospholipid antibody syndrome. AB - Antiphospholipid antibody syndrome (APS) has been shown to be associated with arterial and venous thromboses, recurrent miscarriages, and thrombocytopenia. We describe 3 cases of adult respiratory distress syndrome (ARDS) in patients with primary APS. The autopsy in one patient revealed no evidence of vasculitis, but extensive small vessel thrombosis of multiple organs including the lungs was seen. No infectious process, connective tissue diseases, or pulmonary edema could be documented and there was no clinical or laboratory evidence of vasculitis in the other 2 patients. The 2 patients who survived responded dramatically to intravenous steroids. PMID- 8230029 TI - Myasthenia gravis and scleroderma. AB - Myasthenia gravis is an autoimmune disease that is associated with antibodies to acetylcholine receptors. It is associated with other autoimmune diseases such as thyroiditis (10%), systemic lupus erythematosus (2-8%), rheumatoid arthritis (4 7%), Sjogren's syndrome and polymyositis. It is not commonly found with scleroderma. We describe a case of scleroderma developing in a patient 6 years after the onset of myasthenia gravis. HLA-B8/DR3 may play a role in the association of the 2 conditions. PMID- 8230030 TI - Pyomyositis: cervical localization. AB - We describe an uncommon case of cervical pyomyositis in a 60-year-old woman without predisposing factors, admitted for cervicobrachial neuralgia with fever and paravertebral contracture. Ultrasonography and computed tomography (CT) showed a collection in the left longissimus capitis. Ultrasonography guided needle aspiration yielded thick pus, which cultured Staphylococcus aureus. Our patient completely recovered after antibiotherapy. Pyomyositis is rare and most commonly affects rhizomelic muscles (cervical localization is extremely rare) and occurs in patients with predisposing factors. Staphylococcus aureus is responsible for over 85% of cases; ultrasonography and CT scan are useful for abscess localization and aspiration. PMID- 8230031 TI - Focal myositis: a localized form of polymyositis? AB - We describe an additional case of focal myositis presenting as a painful pseudotumor of the lower leg. Surgical biopsy showed typical features of myositis; the disease improved spontaneously, without clinical or biological abnormalities. Focal myositis can remain a localized process, but in some cases it can become an authentic polymyositis. These generalized forms are associated early with elevated erythrocyte sedimentation rate and/or muscular enzymes. PMID- 8230032 TI - Preferential cellular and humoral immune reactivities to native and denatured collagen types IX and XI in a patient with fatal relapsing polychondritis. AB - We describe a patient with histologically confirmed relapsing polychondritis, an episodic systemic disorder. Although the etiology is unknown and its pathogenesis is incompletely understood, there is evidence strongly suggesting immunologically mediated mechanisms. Enzyme linked immunosorbent assays, immunoblotting and cellular immune responses using lymphocyte proliferation assays showed strong parallel humoral and cellular immune reactivities against collagens type IX and XI. There was also a considerable response to collagen type II which, however, was less pronounced compared to collagen type IX and was directed to native epitopes. Our findings demonstrate a highly distinct immune response to minor matrix collagens in a destructive cartilage disease and thus strongly argue against nonspecific anticollagen immune reactions simply representing epiphenomena resulting from cartilage damage. PMID- 8230033 TI - Avascular necrosis of bone in human immunodeficiency virus infection and antiphospholipid antibodies. AB - We describe 3 cases of human immunodeficiency virus (HIV) infected patients with antiphospholipid antibodies (aPL) in serum who developed avascular necrosis (AVN), an association that to our knowledge, has rarely been described. Given that the 3 patients had stopped their intravenous drug addiction 2 years before the clinical picture appeared, and none had any known risk factors for developing AVN, there may be an association, perhaps fortuitous, between HIV infection, the presence of aPL and the development of AVN. PMID- 8230034 TI - Systemic vasculitis following influenza vaccination--report of 3 cases and literature review. AB - Influenza vaccination is a widely accepted practice particularly among the elderly and high risk individuals. Minor and transitory side effects following the vaccination are common while systemic complications are infrequently reported. We describe 3 patients who developed systemic vasculitis following influenza vaccination. With increasing use of influenza vaccination, attention should be drawn to the possible expression of systemic adverse effects such as vasculitis. PMID- 8230035 TI - Fat necrosis. An unusual cause of polyarthritis. AB - Metastatic fat necrosis is a form of lobular panniculitis which together with synovitis is an unusual complication of pancreatic disease. It can precede an attack of pancreatitis. Pancreatic disease should therefore be remembered as a cause of panniculitis. Serum amylase and abdominal ultrasonography should be considered to exclude pancreatic disease in cases of panniculitis of uncertain etiology. We describe a case of metastatic fat necrosis and discuss the appearance on magnetic resonance imaging. PMID- 8230036 TI - Pancreatitis arthritis with periarticular fat necrosis. AB - We describe the case of a 76-year-old man hospitalized for 14 months because of a complicated hip fracture who developed pancreatitis and polyarthritis. He had no evidence of subcutaneous fat necrosis away from his joints and his pancreatitis was virtually asymptomatic otherwise. Polyarthritis is a rare complication of this disorder, and rarer still is polyarthritis without evidence of subcutaneous fat necrosis elsewhere. PMID- 8230037 TI - Sternoclavicular joint infection due to Bacteroides oralis. AB - A 60-year-old man in good health with no history of intravenous drug abuse was hospitalized for sternoclavicular joint pain and swelling. His complicated course required extensive debridement of the sternoclavicular joint. An insidious course, negative blood culture and requirement for surgery is common in infections of this joint. PMID- 8230038 TI - Course of aPL in patients with primary antiphospholipid syndrome before, during and after pregnancy treated with low dose aspirin. PMID- 8230039 TI - Acute exacerbation mimicking "pseudoseptic" arthritis in rheumatoid arthritis could be caused by abrupt discontinuation of glucocorticoid. PMID- 8230040 TI - Upper gastrointestinal bleeding associated with analgesics and NSAID use: a case control study. PMID- 8230041 TI - Increase in plasma thrombomodulin in patients with systemic sclerosis. PMID- 8230042 TI - Application of interferon-gamma containing gel for local treatment of skin ulcers in autoimmune vasculitides and skin changes in progressive systemic sclerosis. PMID- 8230043 TI - Treatment of sexual dysfunction in a patient with systemic sclerosis. PMID- 8230044 TI - Elevated soluble interleukin 2 receptors are associated with inflammatory arthritis but are not helpful in making a specific diagnosis. PMID- 8230045 TI - Pleural fluid cytology as an indicator of an effusion of rheumatoid origin. PMID- 8230046 TI - Antiphospholipid syndrome in patients with ankylosing spondylitis. PMID- 8230047 TI - Pulmonary hypertension in a case of dermatomyositis. PMID- 8230048 TI - Inflammatory myopathy during ranitidine therapy. PMID- 8230049 TI - Salmonella septic arthritis in patients with human immunodeficiency virus infection. PMID- 8230050 TI - High dose immunoglobulin therapy in a case of inclusion body myositis: clinical and immunologic aspects. PMID- 8230051 TI - Tophaceous gout of patella partita. PMID- 8230052 TI - Postpneumonectomy empyema. PMID- 8230053 TI - Orthogeriatric care and its effect on outcome. PMID- 8230054 TI - The unexamined life is no life for a human being. PMID- 8230055 TI - Student assisted audit of drug prescribing. PMID- 8230056 TI - Epilepsy and driving: attitudes and practices among patients attending a seizure clinic. AB - Although the loss of freedom to drive is a serious consequence of the diagnosis of epilepsy, it is unclear how well current regulations are understood and adhered to by patients. Using questionnaires completed anonymously, we assessed attitudes and practices among 104 patients with epilepsy. Seventeen (16%) patients were driving, three illegally. In total, eight (8%) patients had at some stage driven illegally, even though seven admitted to having been warned not to do so. Even among the 14 patients currently driving and eligible to do so, only eight (57%) had informed the licensing authority and six (43%) their insurers. Only 34 (33%) patients showed satisfactory knowledge of current regulations, including seven (50%) of those currently driving and 21 (50%) of 42 patients who expressed a wish to drive in the future. Of these 34, only 14 (41%) expressed complete agreement with the regulations. Thus, a significant proportion of patients with epilepsy has driven illegally or has failed to inform the licensing authority or insurers. Understanding of regulations is poor. Less stringent restrictions have been shown to be safe in other countries and might improve compliance. PMID- 8230057 TI - Is ophthalmic therapy often overlooked in hospital wards? AB - In a prospective study regarding the accuracy of ophthalmic drugs prescriptions of 55 inpatients in two general teaching hospitals, it was found that: only 27.3% of these patients had their ocular therapy correctly prescribed; 38.2% had their eye medications incorrectly prescribed; and 34.5% had no ophthalmic drugs prescribed. The authors conclude that more care during case history taking could prevent such errors. PMID- 8230058 TI - A survey of industries and colorectal cancer screening of employees in the East Midlands of England. AB - The number of industries in the East Midlands of England who were prepared to enroll their employees in a scheme to detect colorectal cancer by faecal occult blood testing was investigated. Company directors were asked if they would supply a list of their employees over the age of 40 years and allow a research team to give a 15 min talk at the workplace. Forty-nine per cent of firms responded to the questionnaire and 27% of businesses were prepared to enroll. There was no difference in mean total employees between firms willing and unwilling to participate (unpaired t-test, P > 0.05). The commonest reason given by managers rejecting the scheme was that they did not believe the workforce would be interested. PMID- 8230059 TI - Proteinase inhibitors in malignancy: therapeutic promise or another white elephant? PMID- 8230060 TI - The portrayal of the physician in non-medical literature--the bedside manner. PMID- 8230061 TI - John Hunter, pathologist. PMID- 8230062 TI - Felix Marti Ibanez-Iberian Daedalus: the man behind the essays. PMID- 8230063 TI - Posterior paroxysmal discharge: an aid to early diagnosis in Lafora disease. AB - Lafora body disease is a rare neurometabolic disorder of autosomal recessive inheritance. Symptoms begin in the second decade with progressive myoclonic epilepsy and survival is unusual beyond the age of 30. We report an electroencephalographic study in four cases of histologically proven Lafora body disease. Posterior epileptiform discharges were found even in the early stages of the disease and may assist in early diagnosis. PMID- 8230064 TI - Bilateral hip arthroplasties following cardiac transplantation. PMID- 8230065 TI - Synchronous occurrence of an intermediate small cell and a bronchiolo-alveolar carcinoma of the lung. PMID- 8230066 TI - Reno-colic fistula. PMID- 8230067 TI - Pernicious anaemia and multiple gastric polyps with early gastric cancer. PMID- 8230068 TI - Villous adenoma of the common bile duct. PMID- 8230069 TI - Patient confidentiality. PMID- 8230070 TI - Use of clonazepam and valproate in patients with Lance Adams syndrome. PMID- 8230071 TI - Adrenal apoplexy. PMID- 8230072 TI - Prevalence of brucellosis among abattoir workers in Saudi Arabia. AB - The prevalence of brucellosis among abattoir workers in Saudi Arabia was determined through a randomised multi-stage sampling of 1200 abattoir workers. Diagnosis was made by both blood culture and standard tube agglutination test. The overall prevalence of brucellosis was 4.0% among abattoir workers. Infection was more common among butchers (8.9%), veterinarians and veterinary assistants (5.4%), and administrative personnel (1.1%). In order to reach optimum planning for a national brucellosis control programme in Saudi Arabia, ministerial co ordination should be established with formulation of inter-governmental veterinary agreements between the Saudi government and the relevant authorities of animal exporting countries. A national survey on animal marketing is required. PMID- 8230073 TI - The growth pattern of Saudi Arabian pre-school children in Riyadh compared to NCHS/CDC reference population. AB - This study compares the growth patterns of Saudi Arabian pre-school children aged 0-5 years to a reference population of American children documented by the National Center for Health Statistics/Centers for Disease Control (NCHS/CDC). The anthropometric data of weight, height and age were collected in a cross-sectional study of Saudi Arabian children in Riyadh, the capital of Saudi Arabia. The sample selection was accomplished by a 3-stage stratified random procedure, basing the stratification on location and a socio-economic index. A total of 3,795 children comprising 55.2% males and 44.8% females had complete basic anthropometric measurements among other variables. Analyses were effected by using the WorHealth Organization/Centers for Disease Control (WHO/CDC) standard software programme for calculating nutritional indicators. A combination of height for age (as an indicator of stunting) and weight for height (as an indicator of wasting) showed that a sizeable proportion (78.1%) have normal or slightly above normal nutritional status. A small proportion of the children (1.4%) were below -2.0 SD scores of the reference population for weight for height and for age. Overall, Saudi Arabian children are slightly shorter and thinner than their American counterparts. An important finding in weight for height was that nearly 20% of the children were below -2 SD scores of the reference population median. However, due to the very high per capita income, and the fact that food and other essential items are subsidised by the government, the deficits from this reference population may be attributed to inadequate health education nutrition programmes, genetic and other social factors including reproductive behaviour, rather than malnutrition. PMID- 8230074 TI - Knowledge about HIV and views about HIV testing among students at an inner London university. AB - Undergraduate students at an inner London university were surveyed at the beginning of 1992 about their basic knowledge regarding HIV/AIDS, HIV transmission and views about the HIV antibody test. Although they demonstrated good knowledge about HIV/AIDS and routes of transmission, a proportion had had unprotected intercourse, but were deterred from seeking an HIV test for a number of reasons. Implications for promoting safer sex and increasing awareness about HIV testing sites on university campuses are discussed. PMID- 8230075 TI - A quantitative assessment of the Nigerian mothers' ability to prepare salt-sugar solution for the home management of diarrhoea. AB - This study aimed at assessing qualitatively and quantitatively the ability of Nigerian mothers to prepare salt-sugar solutions (SSS) (according to the Nigerian standard formula) under the usual home environment. Mothers were provided with the ingredients but not with measurement spoons nor containers. 274 mothers randomly selected from a peri-urban community participated in the study. Of the 192 (70.1%) who claimed knowledge on SSS preparation, only 47 (24.5%) gave a correct description of its constitution and 103 (54.2%) were willing to prepare the solution. Of the 103 who prepared the solution, 34 (33.0%) used the correct number of teaspoons of salt and of sugar. The composition of the solutions prepared by the mothers varied greatly with sodium levels ranging from 0-760 mmols/l (mean 225.8 +/- 155.3, median 177.3) and glucose, 0-262.6 mmols/l (mean 68.7 +/- 54.4, median 52.0). Only 7 mothers (6.8%) prepared solutions with acceptable sodium and glucose levels. It is concluded that salt-sugar solutions prepared by Nigerian mothers are not safe. There is a great need to review the oral rehydration therapy (ORT) promotion strategies and messages in order to avoid the dangers associated with improperly constituted solutions. PMID- 8230076 TI - Early childhood mortality--a rural study. AB - The present study was conducted in 9 villages of Rural Health Training Centre, Jawan, Aligarh, India, having 1792 registered families. The infant mortality rate was 79.3 per 1000 live births. Higher mortality in children between 1-2 years (29.6/1000) in comparison to 2-5 years (16.2/1000) reflected the higher vulnerability of children below 2 years. Diarrhoea (21.2%) and neonatal factors (21.2%) were the major cause of infant mortality followed by pneumonia (18.2%) and tetanus (15.1%). Diarrhoea (32.2%), pneumonia (22.6%) and malnutrition (12.9%) were the major killers in children between 1-5 years. Mortality in females was higher than males in infancy. Risk factors associated with infant mortality included extremes of maternal age (< 20+ > 35 years) primipara or multipara and illiterate mothers. PMID- 8230077 TI - New trends of postgraduate education in public health in Slovakia (the first experiences of the school of public health in Bratislava). AB - The School of Public Health (SPH) in Bratislava was established on 1 July 1991, as an organisational unit of the Post-graduate Medical School, directly subordinated to the Ministry of Health of the Slovak Republic. The SPH performs both postgraduate and continuing education of specialists in individual public health branches. It participates in the research activities within the field of public health. Our SPH is engaged in the network of European schools of public health, in association with the Association of Schools of Public Health in the European Region (ASPHER). In coordination with these schools, the SPH elaborates on a system of training Europe-wide a comparable qualification degree in Master of Public Health (MPH). In collaboration with the World Health Organization (WHO), the SPH also organizes training courses and seminars with international participation. The SPH entered into the school year 1992/93 with its 5 departments and sub-departments in the following organisational structure: 1. Department of Health Strategy with sub-departments: Health Legislation Health Education Health Sociology 2. Department of Medical Informatics 3. Department of Health Management 4. Department of Medical Education 5. Department of Preventive Medicine with sub-departments: Epidemiology Hygiene of Children and Adolescents Environmental Hygiene Nutrition Preventive Occupational Health Health Protection against Ionization PMID- 8230078 TI - Hair loss in children. PMID- 8230079 TI - The new US health care plan of 1993 and its terminology. AB - The Clinton Administration is moving toward a fundamental change in the United States health care system. President Bill Clinton made the issue of health care reform one of his top campaign priorities and promised to introduce a reform proposal for consideration in 1993. To that end, he asked his wife, Hillary Clinton, to lead the newly established Health Care Reform Task Force as it develops alternatives and the plan for health reform as promised to be introduced in 1993. While some uncertainty exists regarding how President Clinton will differ from candidate Clinton, current indications are that the health care reform will be somewhere between "incremental reform' which the opposition Republican Party favours, and the 'Single Payor/Canadian Style' approaches under which the federal government would take over most responsibilities now carried out by private health insurance companies. The purpose of this paper is to present an overview of the health reform plan, and to present the specialised terminology that is growing from the health reform initiatives. PMID- 8230080 TI - The US health care system: its problems and prospects under the Clinton Administration. PMID- 8230082 TI - Accident prevention and older people: achieving accident prevention. PMID- 8230081 TI - Improving accident prevention for older people. PMID- 8230083 TI - Public health in the United States. PMID- 8230084 TI - RAA system and cardiovascular control in normal subjects, hypertensives and patients with congestive heart failure. AB - The involvement of the renin-angiotensin-aldosterone (RAA) system, particularly angiotensin II, in the pathogenesis of hypertension is widely acknowledged and is supported by several observations: the RAA system has been shown to be critically involved in the development of some experimental hypertensions; activation of the RAA system appears to be the crucial factor involved in the maintenance of the BP elevation in some antihypertensive patients; while drugs which interfere with the production of angiotensin II reduce BP in a large number of hypertensive patients. It is now clear that the chronic BP elevations caused by circulating (and perhaps locally produced) angiotension II may have adverse effects on organ function and protection: for example, induction of cardiac hypertrophy and vascular hypertrophy and/or hyperplasia, reduction of arterial compliance and reduction in vagal tone and facilitation of sympathetic activity on cardiac and vascular targets. At the cardiac level, the renin-angiotensin sympathetic interaction may enhance electrical instability, thereby favouring arrhythmias and increasing mortality after a myocardial infarction. It finally enhances coronary vasoconstriction in man, producing or favouring myocardial ischaemia. PMID- 8230085 TI - New perspectives in angiotensin system control. AB - The 'angiotensin system' is expressed at the whole body, organ/tissue and cellular levels through the action of angiotensin II at specific receptors. An appreciation of the full scope of the actions of angiotensin II (endocrine, paracrine and autocrine) has been made possible by the discovery of the non peptide angiotensin II receptor antagonists, losartan (DuP 753/MK954)(AT1 selective) and PD123177 (AT2-selective). Virtually all of the known effects of angiotensin II are blocked by losartan and designated AT1. Selective AT1 receptor blockade with losartan lowers BP in angiotensin II-dependent models of hypertension, reduces cardiac hypertrophy, improves haemodynamics in models of cardiac failure and reduces the intimal response to vascular injury. AT2 sites have been localised in distinct parts of the brain and in foetal tissue. The functional role of the AT2 sites remains controversial, but possible roles in neuronal ion channel function and collagen metabolism in fibroblasts have been reported. AT1 (losartan-sensitive) receptor subtypes have now been cloned from several rat tissues, suggesting that selective agents of the future may be even more specifically targeted. New perspectives in the control of the angiotensin system continue to evolve rapidly as the new receptor antagonists and molecular biology techniques expand our understanding of angiotensin II. PMID- 8230086 TI - Vascular RAA system. AB - Biochemical and bioassay evidence has proved the existence of tissue RAA systems although their function has not been satisfactorily defined. There are two sources of tissue renin: uptake from plasma and local synthesis. The uptake system has been demonstrated in arterial tissue. Retention of renin can be demonstrated in the aortic wall and the presence of renin at this site is closely correlated with the persistent elevation of blood pressure. Renin gene expression can be demonstrated in several organs such as the liver, brain and arterial wall, although the function of renin or pro-renin at these sites is unknown. Intracellular angiotensin II receptors have been identified that play a role in regulating gene expression. In addition to raising vascular tone, angiotensin II generated in this way could have a trophic action upon cardiac and vascular structures. There is now very strong evidence in favour of the extrarenal RAA system having a pathogenetic role in some forms of hypertension. Renin gene polymorphisms co-segregate with blood pressure in some genetic models, despite normal or low plasma renin and incorporation of an additional mouse renin gene construct into the rat genome produces severe hypertension despite suppression of renal renin. PMID- 8230087 TI - Clinical experience with angiotensin II receptor antagonists. AB - The new generation of antihypertensive agents, the angiotensin II receptor antagonists, lower blood pressure by intervening in the RAA system. Angiotensin II is the major effector compound in the RAA cascade; therefore, blockade of the angiotensin II receptor represents a particularly effective way of preventing its hypertensive effects. Losartan (DuP 753, MK954) is the first of a new series of non-peptide orally-active angiotensin II receptor antagonists. Preliminary results of ongoing pharmacology and efficacy studies with losartan indicate that it may have an improved clinical profile when compared with other antihypertensive agents. The results of these studies have indicated that the effective antihypertensive dose of losartan is 50 mg once daily. PMID- 8230088 TI - Role of angiotensin in the extravascular system. AB - Angiotensin receptors are present in many tissue types, including adrenal cortex, renal glomeruli, heart, hypothalamus, liver, pancreas, pituitary, platelets, renal tubules, uterus and vascular smooth muscle. Two high-affinity receptor subtypes have been identified by radioligand binding with antagonists: losartan (DuP 753/MK954) identifies AT1 receptors; PD123177 and CGP42112A are markers for AT2 receptors. Angiotensin II may be produced locally in tissues outside the humoral system. For example, it is found in the brain, kidney and heart. Within the brain, the heptapeptide angiotensin(1-7) mimics some effects of angiotensin II, but may be formed directly from angiotensin I. Evidence for non-ACE-mediated angiotensin II production has been reported in the heart. Intravascular angiotensin II receptors are implicated in the central release of vasopressin and other hypophyseal hormones, in increasing sympathetic outflow, in the thirst response and, possibly, in cognitive function; in the inotropic and chronotropic effects of angiotensin II on the heart as well as in growth/hypertrophy; in the control of aldosterone release and in the balance between cortisol and aldosterone secretion; and in modulating sodium, chloride and bicarbonate transport within the kidney. Effects on the reproductive system, liver and pancreas have not been established. The pharmacological effects of angiotensin II antagonists will depend on their distribution characteristics as well as affinity for specific receptor subtypes. At present, however, the physiological role of AT2 receptors has not been defined. PMID- 8230089 TI - Concepts and progress in the development of peptide mimetics. PMID- 8230090 TI - Pyrrolo[1,2-a]benzimidazole-based aziridinyl quinones. A new class of DNA cleaving agent exhibiting G and A base specificity. AB - Pyrrolo[1,2-a]benzimidazole(PBI)-based aziridinyl quinones cleave DNA under reducing conditions specifically at G + A bases without any significant cleavage at C + T bases. The postulated mechanisms involve phosphate alkylation by the reductively activated aziridine to afford a hydrolytically labile phosphotriester as well as the classic N(7) purine alkylation followed by depurination and backbone cleavage. Evidence is presented that the phosphate alkylation mechanism could contribute. The PBIs possess a unique spectrum of cytotoxicity against cancer cells (inactive against leukemia but active against nonsmall cell lung, colon, CNS, melanoma, ovarian, and renal cancers). Also reported are results of in vivo antitumor activity screens. PMID- 8230091 TI - Solution conformation of the antitumor drug streptonigrin. AB - The solution conformation of the antitumor drug streptonigrin in THF-d8 has been determined by dynamic 1H NMR spectroscopy (400 MHz). The major solution conformation agrees with the structure observed in the solid state [Chiu, Y.-Y.; Lipscomb, W. N. J. Am. Chem. Soc. 1975, 97, 2525-30]. Rings A, B, and C are coplanar, with ring C held in place by a hydrogen bond from the amino group on ring C and the pyridyl nitrogen in ring B. This conformation is stable in the range pH 3.9-8.9. At lower pH, the hydrogen bond is disrupted due to protonation of the pyridyl nitrogen in ring B. The major species present at pH 3.9-8.9 and 180 K is the zwitterion 1b (80%). Below 190 K, slow proton transfer between the free acid 1a and the zwitterion 1b is observed on the NMR time scale. Addition of a catalytic amount of base to the solution increases the rate of exchange 1a<- >1b, and only one set of resonances is observed. In CD2Cl2 this proton transfer is not observed. Implications for the structure(s) of metal complexes formed by streptonigrin are discussed. PMID- 8230092 TI - Synthesis of A-ring fluorinated derivatives of (17 alpha,20E/Z) [125I]iodovinylestradiols: effect on receptor binding and receptor-mediated target tissue uptake. AB - We have prepared a series of 2- and 4-fluoro derivatives of the isomeric (17 alpha,20E)- and (17 alpha, 20Z)iodovinylestradiols (IVE2) and also the analogs substituted with either a 7 alpha-methyl (7 alpha-Me-IVE2) or 11 beta-methoxy group (11 beta-OMe-IVE2) and evaluated their in vitro and in vivo properties. Electrophilic substitution of the estrone derivatives with N-fluoropyridinium salt gave the 2- and 4-fluoro analogs which were subsequently converted to the 17 alpha-ethynyl derivatives. The tributylstannyl intermediates were obtained from the corresponding 17 alpha-ethynyl analogs using azobisisobutyronitrile or triethylborane as catalyst. All 12 products were also prepared as their no carrier-added [125I]iodovinyl analogs via destannylation of the tributylstannyl precursors. Binding affinity for the estrogen receptor (ER) was in general higher for the 4-F derivatives as compared to the 2-F derivatives, while the 20Z isomers of the same compounds showed somewhat higher ER binding affinity as compared to the 20E isomers. The combination of an A-ring fluoro and 7 alpha- or 11 beta substituent decreased ER binding affinity. Substitution of a fluoro atom at C-4 on either the 17 alpha-ethynylestradiol or isomeric 17 alpha-IVE2 enhanced the affinity of the parent molecule for the ER. A-ring fluorination of all other analogues tested had no effect or depressed ER binding affinity. Varying incubation conditions showed substantial differences in ER binding kinetics between the 20E and 20Z isomers. Tissue distribution in immature female rats showed that the highest uterus uptake and uterus to blood/nontarget ratios in the IVE2 series were obtained with the 4-F-(17 alpha,20Z)IVE2 isomer. The combination of A-ring fluoro and 7 alpha- or 11 beta-substitution decreased uterus uptake but had little or no effect on uterus to blood/nontarget ratios. The highest uterus to blood ratios were observed for the 4-F-(17 alpha,20E)11 beta-OMe-IVE2 (75 at 6 h and 125 at 12 h pi) reflecting rapid blood clearance and in vivo stability, as confirmed by the low levels of thyroid radioactivity. The lack of correlation between ER binding affinities and uterus uptake, and/or uptake ratios, suggests that other factors, including nonspecific binding and metabolic processes, also are involved in the tissue localization process. Our data suggest that 4-F substitution onto (17 alpha,20Z)IVE2 and (17 alpha,20E)11 beta-OMe-IVE2 enhances the potential of these compounds to function as SPECT imaging agents of ER-rich tissues. PMID- 8230093 TI - Conformationally restrained analogs of sympathomimetic catecholamines. Synthesis, conformational analysis, and adrenergic activity of isochroman derivatives. AB - In previous papers dealing with the study of the conformations and the biopharmacological activity of conformationally restrained analogs of sympathomimetic catecholamines (NE and ISO), proposals were advanced for the three-dimensional molecular models A, B, and C; these models provided information about the steric requirements for the direct activation of alpha 1, alpha 2,beta 1, and beta 2 adrenoceptors, respectively. The 1-(aminomethyl)-6,7 dihydroxyisochromans 11 and 12 and the 1-(aminomethyl)-5,6-dihydroxyisochromans 13 and 14 (1-AMDICs) are two different types of semirigid analogs of NE and ISO. The alpha 1, alpha 2, beta 1, and beta 2 adrenergic properties of the 1-AMDICs 11 14 were evaluated in vitro, both by radioligand binding assays and by functional tests on isolated preparations, and were compared with those of their parent compounds (NE and ISO). The results of a conformational study carried out by means of both 1H NMR spectrometry and theoretical calculations indicated that, in these 1-AMDICs, the presumed active groups (aryl moiety, amine nitrogen and benzylic ethereal oxygen) are in a spatial relationship corresponding to the one found for NE and ISO in their preferred conformations, which also proved to be the pharmacophoric conformation in the models A-C. By means of a comparison of the stereostructures of the 1-AMDICs 11-14 with their biopharmacological properties, it was possible to obtain a further definition of the model B with respect to the activation of the alpha 2 adrenoceptors; the superimposition of the 1-AMDICs 11 and 12 with the molecular model C made it possible to detect an area of the beta-adrenergic receptors which might hinder the fit of adrenergic drugs that are analogs of catecholamines with these receptors. PMID- 8230094 TI - Catalytic functionalization of polymers: a novel approach to site specific delivery of misoprostol to the stomach. AB - The application of functionalized polymers to site-directed delivery of the antiulcer prostaglandin, misoprostol, is described. By use of homogeneous catalysis, the simple polymer, polybutadiene, was modified to incorporate the specialized requirements for controlled delivery of misoprostol to the stomach. An acid labile silyl ether bond to the C-11 hydroxyl of misoprostol was installed as the primary rate determining step for drug release, and a series of analogs, in which the steric hindrance about the silicon atom was varied, was prepared and evaluated for in vitro release rates, efficacy against indomethacin induced gastric damage and diarrheagenic activity. The diisopropylsilyl analog, the slowest releasing system studied, showed efficacy equal to misoprostol against indomethacin-induced gastric damage and no diarrhea at the highest dose tested. PMID- 8230095 TI - Structural modification of 5-aryl-2,3-dihydroimidazo[2,1-a]isoquinoline platelet activating factor receptor antagonists. AB - In an effort to determine the effect of modification of the imidazo[2,1 a]isoquinoline portion of the PAF-receptor antagonist SDZ 64-412 (1), several new analogs were prepared and evaluated in vitro and in vivo. One of these, 5-[4-[2 (3,4,5-trimethoxyphenyl)ethyl]phenyl]-2,3-dihydroimidazo [1,2-a]thieno[2,3 c]pyridine (6) was 4-5 times more potent than 1 in inhibiting PAF-induced bronchoconstriction and hemoconcentration when administered po to the guinea pig. PMID- 8230096 TI - 2,4-Diaminothieno[2,3-d]pyrimidine analogues of trimetrexate and piritrexim as potential inhibitors of Pneumocystis carinii and Toxoplasma gondii dihydrofolate reductase. AB - A series of eight previously undescribed 2,4-diaminothieno[2,3-d]pyrimidine analogues of the potent dihydrofolate reductase (DHFR) inhibitors trimetrexate (TMQ) and piritrexim (PTX) were synthesized as potential drugs against Pneumocystis carinii and Toxoplasma gondii, which are major causes of severe opportunistic infections in AIDS patients. 2,4-Diamino-5-methyl-6 (aryl/aralkyl)thieno[2,3-d]pyrimidines with 3,4,5-trimethoxy or 2,5-dimethoxy substitution in the aryl/aralkyl moiety and 2,4-diamino-5 (aryl/aralkyl)thieno[2,3-d]pyrimidines with 2,5-dimethoxy substitution in the aryl/aralkyl moiety were obtained by reaction of the corresponding 2-amino-3 cyanothiophenes with chloroformamidine hydrochloride. The aryl group in the 5,6 disubstituted analogues was either attached directly to the hetero ring or was separated from it by one or two carbons, whereas the aryl group in the 5 monosubstituted analogues was separated from the hetero ring by two or three carbons. 2-Amino-3-cyano-5-methyl-6-(aryl/alkyl)thiophene intermediates for the preparation of the 5,6-disubstituted analogues were prepared from omega-aryl-2 alkylidene-malononitriles and sulfur in the presence of a secondary amine, and 2 amino-3-cyano-4-(aryl/aralkyl)thiophene intermediates for the preparation of the 5-monosubstituted analogues were obtained from omega-aryl-1-chloro-2 alkylidenemalononitriles and sodium hydrosulfide. Synthetic routes to the heretofore unknown ylidenemalononitriles, and the ketone precursors thereof, were developed. The final products were tested in vitro as inhibitors of DHFR from Pneumocystis carinii, Toxoplasma gondii, rat liver, beef liver, and Lactobacillus casei. A selected number of previously known 2,4-diaminothieno[2,3-d]pyrimidines lacking the 3,4,5-trimethoxyphenyl and 2,5-dimethoxyphenyl substitution pattern of TMQ and PTX, respectively, were also tested for comparison. None of the compounds was as potent as TMQ or PTX, and while some of them showed some selectivity in their binding to Pneumocystis carinii and Toxoplasma gondii versus rat liver DHFR, this effect was not deemed large enough to warrant further preclinical evaluation. PMID- 8230098 TI - A series of penicillin derived C2-symmetric inhibitors of HIV-1 proteinase: synthesis, mode of interaction, and structure-activity relationships. AB - The C2-symmetric diester 1 was identified by random screening as a novel inhibitor of HIV-1 proteinase. This led to the preparation of a series of related more potent amides from readily accessible penicillins. Many of the compounds showed potent antiviral activity in HIV-1-infected MT-4 cells and an ability to inhibit syncytia formation in infected C8166 cells, with no evidence of cytotoxicity. The compounds showed no activity against other aspartyl proteinases (renin, pepsin, and cathepsin D). Structure-activity relationships support a symmetrical interaction with the enzyme. Pharmacokinetic evaluation of the ethylamide 3 revealed it was subject to rapid plasma clearance and had low oral bioavailability. PMID- 8230097 TI - A series of penicillin-derived C2-symmetric inhibitors of HIV-1 proteinase: structural and modeling studies. AB - The binding modes of a series of penicillin-derived C2 symmetric dimer inhibitors of HIV-1 proteinase were investigated by NMR, protein crystallography, and molecular modeling. The compounds were found to bind in a symmetrical fashion, tracing and S-shaped course through the active site, with good hydrophobic interactions in the S1/S1' and S2/S2' pockets and hydrogen bonding of inhibitor amide groups. Interactions with the catalytic aspartates appeared poor and the protein conformation was very similar to that seen in complexes with peptidomimetics, in spite of the major differences in ligand structure. PMID- 8230099 TI - Synthesis and structure-activity relationships of a series of penicillin-derived HIV proteinase inhibitors containing a stereochemically unique peptide isostere. AB - A series of HIV-1 proteinase inhibitors was synthesized based upon a single penicillin derived thiazolidine moiety. Reaction of the C-4 carboxyl group with (R)-phenylalaninol gave amide 10 which was a moderately potent inhibitor of HIV-1 proteinase (IC50 = 0.15 microM). Further modifications based on molecular modeling studies led to compound 48 which contained a stereochemically unique statine-based isostere. This was a potent competitive inhibitor (Ki = 0.25 nM) with antiviral activity against HIV-1 in vitro (5 microM). Neither modification to the benzyl group in an attempt to improve interaction with the S2' pocket, nor introduction of a hydrogen bond donating group to interact with residue Gly48' resulted in improved inhibitory or antiviral activity. PMID- 8230100 TI - Azulenic retinoids: novel nonbenzenoid aromatic retinoids with anticancer activity. AB - Several novel azulene-containing retinoids were prepared and evaluated for their ability to suppress carcinogen-induced neoplastic transformation and to concomitantly up-regulate gap junctional communication in the in vitro mouse fibroblast C3H/10T1/2 cell bioassay. The azulenic retinoids were divided into two groups: compounds 1-6 were modeled after retinoic acid with flexible polyenic side chain whereas retinoids 7-13 featured a benzoic acid moiety analogous to the prototypic retinobenzoate (E)-4-[2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2- naphthalenyl)-1-propenyl]benzoic acid (TTNPB). Within this latter group the side chains for compounds 7, 10, and 11 were attached at the 1-, 2-, and 8-positions of the azulenic terminus, respectively. Biological activities were determined for all the new compounds. Two of these novel retinoids, azulenic retinobenzoic acid derivatives 7 and 11, were completely effective inhibitors of transformation at 10(-6) M. The most active azulenic retinoids also enhanced gap junctional communication in untransformed cells; this was associated with up-regulated expression of connexin 43, a structural protein of the gap junction. Two fluorinated analogs were also tested. The azulenic fluoro acid 5 was found to be more potent than the trifluoromethyl analog 6. Azulenic analogs with hydroxyl or carboxaldehyde side chain functional groups were ineffective transformation inhibitors. In general, azulenic retinobenzoic acid analogs structurally akin to TTNPB were more effective than flexible side chain analogs related to retinoic acid. PMID- 8230101 TI - Synthesis and antibacterial activity of a new series of tetracyclic pyridone carboxylic acids. AB - A novel tetracyclic pyridone carboxylic acid with a thiazolidine ring, 1,2 dihydro-9,1-(epoxymethano)-7-fluoro-8-(4-methyl-1-piperazinyl)-5-oxo -5H- thiazolo[3,2-a]quinoline-4-carboxylic acid (4a), and variants with a nitrogen atom (4b) or carbonyl group (4c) in the place of the 10-position oxygen atom of 4a were prepared and tested for antibacterial activity and inhibitory activity on DNA gyrase from Escherichia coli KL-16. The in vitro antibacterial potency with regard to the 10-position atom was found to be of the following order; O > NCH3 = C = O. The IC50 values for DNA gyrase inhibition activity for the 4a, 4b, and 4c compounds were 0.33, 0.53, and 0.67 g/mL, respectively. The activity of 4a, in which the C-3 methyl group and C-5 of ofloxacin (2a) were connected with a sulfur atom to restrict the conformation of 2a, was more potent than that of 2a against both Gram-positive and -negative bacteria, except for Pseudomonas aeruginosa. Compared to the tetracyclic pyridone carboxylic acid 1a, which has a flat thiazole ring, compound 4a showed comparable or slightly more potent activity against both Gram-positive and -negative bacteria, except for P. aeruginosa. PMID- 8230102 TI - Synthesis of (R,S)-trans-8-hydroxy-2-[N-n-propyl-N-(3'-iodo-2' propenyl)amino]tetral in (trans-8-OH-PIPAT): a new 5-HT1A receptor ligand. AB - In order to develop tracers with higher specific activity to supplant the currently used [3H]-8-OH-DPAT [8-hydroxy-2-(N,N-di-n-propylamino)tetralin] for in vitro and in vivo evaluation of 5-HT1A receptors, a new radioiodinated ligand was prepared. (R,S)-trans-8- Hydroxy-2-[N-n-propyl-N-(3'-iodo-2' propenyl)amino]tetralin (trans-8-OH-PIPAT), 8, was synthesized by a 10-step reaction. Binding studies with rat hippocampal membrane homogenates showed that 8 exhibited a Ki value of 0.92 nM against (R,S)-[3H]-8-OH-DPAT. Radiolabeled [125I] 8 was prepared from the corresponding tri-n-butyltin precursor via an oxidative iododestannylation reaction with sodium [125I]iodide. Binding studies in the hippocampal homogenates revealed that [125I]-8 bound to a single high-affinity site (Kd = 0.38 +/- 0.03 nM,Bmax = 310 +/- 20 fmol/mg of protein). Competition binding experiments clearly indicated that the new ligand displayed the expected 5-HT1A receptor binding profile. The rank order of potency was (R,S)-trans-8-OH PIPAT > (R,S)- 8-OH-DPAT > WB4101 > 5-HT > (R,S)-trans-7-OH-PIPAT > (R,S)-7-OH DPAT > (R,S)-propranolol > spiperone >> ketanserin >> dopamine > atropine. This new ligand offers several unique advantages, including high specific activity, high binding affinity, and low nonspecific binding, all of which make it an excellent probe for the investigation and characterization of 5-HT1A receptors. PMID- 8230103 TI - The interaction of ammonium, sulfonium, and sulfide analogues of metoclopramide with the dopamine D2 receptor. AB - A series of permanently charged ammonium and sulfonium analogues of metoclopramide as well as a permanently uncharged sulfide analogue were synthesized and evaluated for their ability to inhibit apomorphine-induced responses on mouse striatal slices. Three of the four permanently charged analogues were found to inhibit apomorphine's effects, although at higher concentrations than either metoclopramide or its dimethyl analogue. In contrast, the sulfide analogue was inactive at concentrations up to 100 microM. These findings are consistent with earlier studies of chlorpromazine and sulpiride analogues and provide further evidence that dopamine antagonists bind in their charged molecular forms to anionic sites on the D2 receptor. Further, the results of this study in conjunction with those of our earlier sulpiride study would seem to indicate that differences in the biological profiles of metoclopramide, a type 1 benzamide useful as a gastric prokinetic agent, and sulpiride, a type 2 benzamide useful for its antipsychotic effects, are not due to any appreciable differences in the binding of the basic nitrogen atom of these molecules. PMID- 8230104 TI - Voronoi modeling: the binding of triazines and pyrimidines to L. casei dihydrofolate reductase. AB - Vorom is a computer-aided method of drug design which can model a biological receptor given only binding data of known ligands. Using the binding energies of known competitive, reversible ligands of a biological macromolecule, vorom can make predictions about the binding energies and conformations of other small molecules binding to that receptor as well as provide information about the geometry and physicochemical characteristics of the binding site. One such model of L. casei dihydrofolate reductase was made. The model was able to predict the binding energies of 31 pyrimidine and triazine inhibitors out of a total set of 47, using only eight of the molecules (four pyrimidines and four triazines) as input. The binding energy of methotrexate, which is neither a pyrimidine nor a triazine, was correctly predicted. The binding mode of methotrexate predicted by vorom is entirely consistent with known X-ray data. The predicted binding modes of the pyrimidine inhibitors and the geometry of the site model are also consistent with published NMR and crystallographic data. PMID- 8230105 TI - Efficient inhibition of human leukocyte elastase and cathepsin G by saccharin derivatives. AB - A series of saccharin derivatives I has been synthesized and evaluated for their inhibitory activity toward human leukocyte elastase and cathepsin G. Most of the compounds were found to be efficient and time-dependent inhibitors of elastase. Inactivated elastase was found to regain its activity almost fully after 24 h (80 90% activity) and the half-lives of reactivation ranged between 12-15 h. Addition of hydroxylamine to fully-inactivated enzyme led to rapid and complete recovery of enzymatic activity. A tentative mechanism of action is proposed on the basis of biochemical and model studies. PMID- 8230106 TI - TIPP[psi]: a highly potent and stable pseudopeptide delta opioid receptor antagonist with extraordinary delta selectivity. AB - Pseudopeptide analogues of the delta opioid antagonists H-Tyr-Tic-Phe-Phe-OH (TIPP) and H-Tyr-Tic-Phe-OH (TIP) containing a reduced peptide bond between the Tic2 and Phe3 residues were synthesized. The two compounds, H-Tyr-Tic psi [CH2NH]Phe-Phe-OH (TIPP [psi]) and H-Tyr-Tic psi-[CH2NH]Phe-OH (TIP [psi]), were tested in mu-, delta-, and kappa-receptor-selective binding assays and in the guinea pig ileum (GPI) and mouse vas deferens (MVD) bioassays. In comparison with their respective parent peptides, both pseudopeptide analogues showed increased delta antagonist potency in the MVD assay, higher delta receptor affinity and further improved delta receptor selectivity. The more potent compound, TIPP [psi], displayed subnanomolar delta receptor affinity and in direct comparisons with other selective delta ligands was shown to have unprecedented delta specificity (Ki mu/Ki delta = 10,500). Furthermore, this compound turned out to be highly stable against enzymatic degradation and, unlike other delta antagonists, showed no mu or kappa antagonist properties. TIPP [psi] is likely to find wide use as a pharmacological tool in opioid research. PMID- 8230107 TI - Substituted 3-phenylpiperidines: new centrally acting dopamine autoreceptor antagonists. AB - The (+)-and (-)-enantiomer of compounds 4 and 5 were synthesized and tested for central dopamine (DA) receptor stimulating activity, using biochemical and behavioral tests in rats. Based on the available data the (-)-enantiomers of 4 and 5 are characterized as centrally acting DA autoreceptor antagonists with oral activity. They display a similar pharmacological profile as the prototype DA autoreceptor antagonists (+)-1 and (+)-2 and show a certain preference for the D3 DA receptor antagonist binding site. PMID- 8230108 TI - Troger's base. An alternate synthesis and a structural analog with thromboxane A2 synthetase inhibitory activity. AB - The synthesis of 2,8-dimethyl-6H,12H-5,11-methanodibenzo[b,f][1,5]diazocine (Troger's base) from p-toluidine and of two Troger's base analogs from other anilines by reaction with hexamethylenetetramine in trifluoroacetic acid is described. 2,3,6,7-Tetrahydro-9-methyl-2,6-di-p-tolyl-1H,5H-pyrimido[5,6,1-ij] quinazoline is formed as a secondary product in the reaction of p-toluidine and hexamethylenetetramine. One of the Troger's base analogs, 2,8-bis(3' pyridylmethyl)-6H,12H-5,11-methanodibenzo[b,f][1,5]d iazocine (5), is an effective inhibitor of the enzyme, thromboxane A2 (TxA2) synthase, with an ED50 of 30 ng/mL in a specified in vitro assay. Three analogs having substituents on the bridging methylene group of the bicyclic nucleus of the Troger's base structure were prepared, but all were considerably less active than the aforementioned compound in the inhibition assay. The structures of these inhibitors of TxA2 synthase fall outside the classical structure-activity relationship that has been established for this class of enzyme inhibitors. PMID- 8230109 TI - A potent, orally active, balanced affinity angiotensin II AT1 antagonist and AT2 binding inhibitor. PMID- 8230110 TI - 3-(Alkylamino)-4H-pyrido[4,3-e]-1,2,4-thiadiazine 1,1-dioxides as powerful inhibitors of insulin release from rat pancreatic B-cells: a new class of potassium channel openers? PMID- 8230111 TI - Structure-activity relationship within a series of pyrazolidinone antibacterial agents. 1. Effect of nuclear modification on in vitro activity. AB - The synthesis and biological evaluation of a series of pyrazolidinone-containing mono- and bicyclic compounds are described. The results of this investigation indicate that the [3.3.0] ring system bearing strongly electron-withdrawing groups in the 3-position provides the optimal arrangement for antibacterial activity. Two highly potent derivatives, LY193239 and LY255262, have been selected for further preclinical evaluation. PMID- 8230112 TI - Structure-activity relationship within a series of pyrazolidinone antibacterial agents. 2. Effect of side-chain modification on in vitro activity and pharmacokinetic parameters. AB - The structure-activity relationship among a series of novel pyrazolidinone antibacterial agents is described. Specifically, the effect of modification of the side chain attached to the nitrogen at C-7 was explored in an attempt to improve the potency and spectrum of activity. This approach was successful in identifying several compounds having good in vitro profiles. These top candidates were then evaluated for their activity in vivo, and their pharmacokinetic behavior in various animal models was explored. This information proved critical for the identification of candidates for clinical evaluation. PMID- 8230113 TI - TSAO analogues. 3. Synthesis and anti-HIV-1 activity of 2',5'-bis-O-(tert butyldimethylsilyl)-beta-D-ribofuranosyl 3'-spiro-5''-(4''-amino-1'',2'' oxathiole 2'',2''-dioxide) purine and purine-modified nucleosides. AB - Several purine and purine-modified analogues of the new lead anti-HIV-1 agent [[2',5'-bis-O-(tert-butyldimethylsilyl)-beta-D-ribofuranosyl] thymine]-3'-spiro 5''-(4''-amino-1'',2''-oxathiole 2'',2''-dioxide) (TSAO-T) have been prepared and evaluated as inhibitors of HIV-1-induced cytopathicity. Reaction of O mesylcyanohydrins of furanos-3'-ulosyladenine with Cs2CO3 afforded beta-D-xylo- and ribofuranosyladenine 3'-spiro nucleosides. Reaction of 1,2-di-O-acetyl-5-O benzoyl-3-C-cyano-3-O-mesyl-D-ribofuranose with purine bases, followed by treatment with Cs2CO3, stereoselectively afforded beta-D-ribofuranosyl 3'-spiro nucleosides. 2',5'-O-Deacylation and subsequent treatment with tert butyldimethylsilyl chloride gave the required TSAO derivatives. The 3'-spiro nucleosides with a xylo configuration did not show any anti-HIV activity. However, the purine ribo 3'-spiro nucleosides were potent and selective inhibitors of HIV-1 with a 50% effective concentration in the range of 0.1-1 microM and a selectivity index ranging from 2 to 3 orders of magnitude. Introduction of an alkyl function at N-1 of the purine moiety markedly decreased cytotoxicity without affecting antiviral activity. PMID- 8230114 TI - Antipicornavirus activity of tetrazole analogues related to disoxaril. AB - A series of tetrazole analogues of Win 54954, a broad-spectrum antipicornavirus compound, has been synthesized to address the acid lability of the oxazoline ring of this series of compounds. The results of X-ray crystallography studies of several members of the oxazoline series bound to human rhinovirus type 1A and 14 have been used to design compounds in the tetrazole series with a broad spectrum of activity. Compound 16b, which has a three-carbon linkage between the isoxazole and phenyl rings and a propyl chain extending from the isoxazole ring, exhibiting an MIC80 for 15 rhinovirus serotypes of 0.20 microM as compared to 0.40 microM for Win 54954. X-ray studies of 16b bound to human rhinovirus-14 show that the propyl side chain extends into a pore in the binding site with the possibility of hydrophobic interactions with a pocket formed by Leu106 and a portion of Ser107. PMID- 8230115 TI - Inhibitors of blood platelet cAMP phosphodiesterase. 4. Structural variation of the side-chain terminus of water-soluble 1,3-dihydro-2H-imidazo[4,5-b]quinolin-2 one derivatives. AB - 1-(Cyclohexylmethyl)-4-[4-[(2,3-dihydro-2-oxo-1H-imidazo[4,5-b] quinolin-7 yl)oxy]-1-oxobutyl]piperazine (2) was previously identified as a potent, water soluble inhibitor of human blood platelet cAMP phosphodiesterase and of induced aggregation in vitro that demonstrated effective antithrombotic activity in animal models of thrombosis. Although 2 exhibited 25% oral bioavailability in rats, pharmacokinetic studies conducted in monkeys revealed that the parent compound was less than 5% bioavailable, the result of extensive first-pass biotransformation in the liver. In an effort to identify potent platelet aggregation inhibitors with enhanced metabolic stability, the side-chain amide moiety of 2 was replaced with chemically more stable urea (6a-s), sulfonamide (13a-m), sulfone (19a-r), and tetrazole (23a-s) moieties. Many representatives from each of these structural types effectively combined potent inhibition of ADP induced human platelet aggregation in vitro with excellent aqueous solubility, and several are superior to 2. Within each series, the N-(cyclohexylmethyl)-, N (2-ethylbutyl)-, N-benzyl-, and N-(4-fluorobenzyl)-substituted derivatives were evaluated for in vitro metabolic stability by incubating with the S-9 fraction of monkey liver for 2 h, and the extent of biotransformation was compared with that of the prototype 2. The sulfone 19e and the tetrazoles 23e, 23g, 23j, and 23q were significantly more stable than 2 under these conditions, and 19e and 23e were selected for evaluation in vivo. Tetrazole 23e exhibited 72% bioavailability following ip administration to rats compared with 35% bioavailability for 2 and 19e under the same conditions. However, the oral bioavailability of 19e and 23e in the rat was estimated to be only 3%, suggesting that 19e and 23e are less readily absorbed from the gastrointestinal tract than 2. PMID- 8230116 TI - Conformational analysis of a highly potent dicyclic gonadotropin-releasing hormone antagonist by nuclear magnetic resonance and molecular dynamics. AB - Structural analysis of constrained (monocyclic) analogues of gonadotropin releasing hormone (GnRH) has led to the development of a model for the receptor bound conformation of GnRH and to the design of highly potent, dicyclic GnRH antagonists. This is one of the first cases where a dicyclic backbone has been introduced into analogues of a linear peptide hormone with retention of high biological activity. Here we present a conformational analysis of dicyclo(4-10,5 8)[Ac-D-2Nal1-D-pClPhe2-D-Trp3-Asp4+ ++-Glu5-D-Arg6-Leu7-Lys8- Pro9-Dpr10]-NH2 (I), using two-dimensional nuclear magnetic resonance (NMR) spectroscopy and molecular dynamics simulation. Compound I inhibits ovulation in the rat at a dose of 5-10 micrograms (Rivier et al. In Peptides: Chemistry, Structure ad Biology; Rivier, J. E., Marshall, G. R., Eds.; ESCOM: Leiden, The Netherlands, 1990; pp 33 37). The backbone conformation of the 4-10 cycle in this dicyclic compound is very similar to that found previously for a parent monocyclic (4-10) GnRH antagonist (Rizo et al. J. Am. Chem. Soc. 1992, 114, 2852-2859; ibid. 2860-2871), which gives strong support to the hypothesis that GnRH adopts a similar conformation upon binding to its receptor. In this conformation, residues 5-8 form a "beta-hairpin-like" structure that includes two transannular hydrogen bonds and a Type II' beta turn around residues D-Arg6-Leu7. The "tail" of the molecule formed by residues 1-3 is somewhat structured but does not populate a single major conformation. However, the orientation of the tail on the same side of the 4-10 cycle as the 5-8 bridge favors interactions between this bridge and the tail residues. These observations correlate with results obtained previously for the parent monocyclic (4-10) antagonist, and have led to the design of a series of new dicyclic GnRH antagonists with bridges between the tail residues and residues 5 or 8. PMID- 8230117 TI - The crystal structure, absolute configuration, and phosphodiesterase inhibitory activity of (+)-1-(4-bromobenzyl)-4-(3-(cyclopentyloxy)- 4-methoxyphenyl) pyrrolidin-2-one. AB - Chiral HPLC resolution of the phosphodiesterase IV (PDE IV) inhibitor rolipram (1) provided (-)-1, and this enantiomer was converted into its 1-(4-bromobenzyl) derivative, (+)-2. X-ray structural analysis of (+)-2 established the absolute configuration as R, which provides the first direct evidence for a previously assumed assignment of configuration. The crystal structure of (+)-2 and the PDE inhibitory activity of both enantiomers of 2 are discussed in the context of a previously proposed topological model. PMID- 8230118 TI - Non-steroidal glucocorticoid-like substances: receptor binding and in vivo activity. AB - Compounds of general structure I, prepared by a Diels-Alder reaction with diene 3, are relatives of the known potent glucocorticoid II but possess a markedly modified C- and D-ring environment. Despite these structural changes, 4, 5, 9, 10, 12a, 13, and 14 bound to the glucocorticoid receptor with an affinity which approximated that of the reference standard, 6-alpha-methylprednisolone. Four of these compounds not only exhibited antiinflammatory activity in the alpha tocopherol pouch test but also exhibited marked adrenal suppression and other typical glucocorticoid properties at doses in the same range as the effective antiinflammatory doses. PMID- 8230119 TI - 5-HT3 receptor antagonists. 3. Quinoline derivatives which may be effective in the therapy of irritable bowel syndrome. AB - A series of quinolinecarboxylic acid derivatives has been previously described as a new class of 5-HT3 receptor antagonists due to deviation of a carbonyl moiety from the place of an aromatic ring in their minimum-energy conformations. These derivatives were evaluated in a wrap-restraint stress-induced defecation model in rats. Reference compounds, ondansetron (1), granisetron (2), and YM060 (4), potently inhibited a stress-induced increase in stools excreted from fed rats (ID50 = 0.27, 0.12, and 0.0052 mg/kg, po, respectively). However, quinoline derivatives exhibited different activities depending on structural class. 4 Hydroxyquinoline-3-carboxylic acid derivatives 5 and 6a possess high affinity for the 5-HT3 receptor (Ki = 6.1 and 1.5 nM, respectively) and exhibit potent activity in the Bezold-Jarisch (B-J) reflex test (ED50 = 0.0017 and 0.000 10 mg/kg, i.v., respectively), but they did not effectively inhibit the increase in fecal pellet output at the dose of 1 mg/kg, po. On the other hand, most of 1 substituted 2-oxoquinoline-4-carboxylates 10 showed less potent activity in the B J reflex test than 1 or 2 but inhibited restraint stress-induced defecation more potently than 1 or 2. The ID50 value of endo-8-methyl-8- azabicyclo[3.2.1]oct-3 yl 1-isobutyl-2-oxo-1,2-dihydro-4- quinolinecarboxylate 10e was 0.013 mg/kg, po. With respect to the selected compounds 6a and 10e, effects of 5-HT- and thyrotropin-releasing hormone (TRH)-induced defecation, castor oil-induced diarrhea and wrap-restraint stress-induced colonic propulsion in rats were examined. These 5-HT3 receptor antagonists did not effectively inhibit castor oil induced diarrhea, which has been reported not to be mediated via the 5-HT3 receptor. Although 10e showed 800-fold decreased potency compared with 4 in the B J reflex test, 10e exhibited activity as potent as 4 in 5-HT- and TRH-induced defecation assays; 10e exhibited 7-fold increased potency compared with 4 in wrap restraint stress-induced colonic propulsions. From these results, 10e appears to interact selectively with 5-HT3 receptors in the gastrointestinal system and might be effective in the therapy of irritable bowel syndrome (IBS). PMID- 8230120 TI - Inhibitors of acyl-Coa:cholesterol acyltransferase. 4. A novel series of urea ACAT inhibitors as potential hypocholesterolemic agents. AB - We have synthesized a series of N-phenyl-N'-aralkyl and N-phenyl-N'-(1 phenylcycloalkyl)ureas as inhibitors of acyl-CoA:cholesterol acyltransferase (ACAT). This intracellular enzyme is thought to be responsible for the esterification of dietary cholesterol; hence inhibition of this enzyme could reduce diet-induced hypercholesterolemia. For this series of compounds, the in vitro ACAT inhibitory activity was improved by increasing the bulk of the 2,6 substituents on the phenyl ring. Additionally, we found that spacing of the aromatic rings was critical for ACAT inhibitory activity. A phenyl ring five atoms away from the requisite 2,6-diisopropylphenyl moiety was optimal for in vitro activity. Substitution alpha to the N'-phenyl moiety enhanced in vitro potency. In the case of phenylcycloalkyl ureas, ACAT inhibitory activity was independent of the size of the cycloalkyl ring. From this series of analogs, compound 25, which had excellent in vitro potency for inhibiting ACAT, was found to lower plasma cholesterol by 73% in vivo when administered in the diet at 50 mg/kg in an animal model of hypercholesterolemia. In this model, compound 25 lowered plasma cholesterol dose dependently and was as efficacious as the Lederle ACAT inhibitor CL 277082. PMID- 8230121 TI - Synthesis of structural analogs of leukotriene B4 and their receptor binding activity. AB - Structural analogs of leukotriene B4 (LTB4) were designed using a preferred conformation of LTB4 (1). Appending an aromatic ring scaffold between LTB4 carbons 7 and 11 led to quinoline analogs 3 and 15. A similar modification to the LTB4 structure between carbons 7 and 9 led to the pyridine analogs 41 and 46. The compounds of this study were evaluated in receptor binding assays using [3H]LTB4 and intact human DMSO differentiated U-937 cells. The first analog prepared, quinoline 3, displayed moderate potency in the LTB4 receptor binding assay (Ki = 0.9 microM). Modification of 3 by appending an aromatic ring between carbons 2 and 4 of the acid side chain produced a dramatic increase in receptor binding (15, Ki = 0.01 microM); a further improvement in receptor binding was achieved in the pyridine series (e.g., 41; Ki = 0.001 microM). The LTB4 receptor agonist/antagonist activity of the test compounds was determined using a functional assay that relies upon intracellular calcium mobilization induced by LTB4. Of the analogs prepared in this report only 47 demonstrated LTB4 receptor antagonist activity. PMID- 8230122 TI - Trisubstituted pyridine leukotriene B4 receptor antagonists: synthesis and structure-activity relationships. AB - A series of trisubstituted pyridines have been prepared that exhibit in vitro leukotriene B4 (LTB4, 1) receptor antagonist activity. Previous disubstituted pyridines from these labs showed high affinity for the LTB4 receptor but demonstrated agonist activity in functional assays (e.g., 2, Ki = 1 nM). Compound 4, the initial lead compound of this new series, showed only modest affinity by comparison (Ki = 282 nM); however, 4 was a receptor antagonist with no demonstrable agonist activity up to 10 microM. Subsequent modifications of the lipid tail and aryl head group region led to the discovery of aniline 50 (SB 201146). This compound, also free of agonist activity, possesses high affinity for the LTB4 receptor (Ki = 4.7 nM). PMID- 8230123 TI - Synthesis and LTB4 receptor antagonist activities of the naturally occurring LTB4 receptor antagonist Leucettamine A and related analogues. AB - The isolation and structure determination of the naturally occurring LTB4 receptor antagonist Leucettamine A (1) was recently reported. Herein we describe the synthesis of this natural product, the preparation of several analogues, and their effectiveness as antagonists of [3H]LTB4 binding to intact human U-937 cells. Total synthesis of Leucettamine A (1) is achieved by a convergent route which takes advantage of the elements of symmetry within the molecule. Syntheses of analogues of 1, which lacked the same degree of symmetry, are achieved by a different approach starting from alpha-amino acids. The natural product 1 inhibits [3H]LTB4 binding to its receptors on intact human U-937 cells with a Ki = 3.5 +/- 0.8 microM and is devoid of measurable agonist activity at the concentrations tested. 2-Amino imidazole analogues of 1 lacking the dioxolane groups were prepared. Generally these are significantly less potent than 1. However, one (26), designed on the basis of a putative structural overlay with LTB4, demonstrated potency comparable to that of the natural product (Ki = 2.4 +/ 0.2 microM). PMID- 8230124 TI - Synthesis of paraxanthine analogs (1,7-disubstituted xanthines) and other xanthines unsubstituted at the 3-position: structure-activity relationships at adenosine receptors. AB - Synthetic procedures for the preparation of various 3-unsubstituted xanthines, including paraxanthine analogs (1,7-disubstituted xanthines) and 1,8 disubstituted xanthines, were developed. Silylation of 1-substituted xanthines followed by alkylation at the 7-position provides a facile route to paraxanthine analogs. Regioselective alkylation of tris(trimethylsilyl)-6-aminouracil provides 3-substituted 6-aminouracils, which are converted to 1,8-disubstituted xanthines by standard procedures. The ring closure of 3-substituted 5 cyclopentanecarboxamido- and 5-(benzoylamino)-6-aminouracils requires drastic reaction conditions. Affinity for brain A1 and A2 adenosine receptors was determined in binding assays for these and other xanthines with substituents in 1 , 3-, 7-, 8-, and 9-positions. Substitution at the 1-position was necessary for high affinity at adenosine receptors. 1,3-Disubstituted xanthines generally had higher affinity than 1,7-disubstituted xanthines. 1,8-Disubstituted xanthines had high affinity for adenosine receptors; some were highly selective for A1 receptors. PMID- 8230125 TI - Synthesis and anticonvulsant activities of alpha-heterocyclic alpha-acetamido-N benzylacetamide derivatives. AB - Earlier studies showed that (R,S)-alpha-acetamido-N-benzylacetamides (2) containing a five- and six-membered aromatic or heteroaromatic group appended at the C(alpha) site displayed outstanding activity in the maximal electroshock induced seizure (MES) test in mice. An expanded set of C(alpha)-heteroaromatic analogues of 2 have been prepared and evaluated. The observed findings extended the structure-activity relationships previously discerned for this novel class of anticonvulsants and have validated previous trends. The alpha-furan-2-yl (4), alpha-oxazol-2-yl (18), and alpha-thiazol-2-yl (19) alpha-acetamido-N benzylacetamides afforded excellent protection against MES-induced seizures in mice. The ED50 and PI values for these adducts rivaled those reported for phenytoin. The outstanding properties provided by 4 led to an in-depth examination of the effect of structural modification at key sites within this compound on biological activity. The pharmacological data in this series indicated that stringent steric and electronic requirements existed for maximal activity and revealed the outstanding activity of (R)-(-)-alpha-acetamido-N-(4 fluorobenzyl)-alpha-(furan-2-yl)aceta mide [(R)-30]. PMID- 8230126 TI - 4,5-Dihydro-1-phenyl-1H-2,4-benzodiazepines: novel antiarrhythmic agents. AB - A series of 4,5-dihydro-1-phenyl-1H-2,4-benzodiazepines has been identified as potential antiarrhythmic agents that interact with sodium and potassium channels and prolong the ventricular effective refractory period (ERP) in anesthetized guinea pigs. Concomitant displacement of radiolabeled bactrachotoxin from site II in Na+ channels and of radiolabeled dofetilide from delayed rectifier K+ channels was evident with all members of this chemical series at a concentration of 10 microM. Structure-activity relationship (SAR) studies using a paced guinea pig model to assess prolongation of the ERP indicated that methyl or ethyl at the 1 position had little effect on activity, while larger groups caused a diminution of activity. Compounds with substituents at either the 3- or 4-position that increased lipophilicity generally were more potent; however, too many lipophilic substituents simultaneously at positions 1, 3, and 4 resulted in less active compounds. Substituents on either aromatic ring had little influence on activity, and phenyl at the 5-position resulted in a significant reduction in antiarrhythmic activity. When two sets of enantiomerically pure compounds were tested in the guinea pig, chirality was shown to be important for activity of 8, where the (R)-enantiomer was the more active, but not in the case of 15, where the enantiomers were equiactive. Several compounds in this series increased the threshold for ventricular fibrillation and refractoriness in myocardially infarcted anesthetized cata and delayed the onset of aconitine-induced arrhythmias in anesthetized guinea pigs following intravenous dosing. Moreover, these compounds possessed oral antiarrhythmic activity in conscious myocardially infarcted dogs. Compound R-15 has been advanced for further biological and toxicological evaluations. PMID- 8230127 TI - A new series of imidazolones: highly specific and potent nonpeptide AT1 angiotensin II receptor antagonists. AB - Starting from the structure of the novel nonpeptide AT1 receptor antagonist DuP 753 (losartan), a new series of potent antagonists was designed. In these compounds the central imidazole nucleus was replaced by the dihydroimidazol-4-one structure. The most active compounds had a spirocyclopentane or a spirocyclohexane ring in position 5. Like the imidazole series, the best substituents were the linear butyl chain in position 1 and the [2'-(tetrazol-5 yl)biphenylyl]methyl group in position 3. Antagonistic activity was assessed by the ability of the compounds to competitively inhibit [125I]AII binding to the AT1 subtype receptor and to antagonize AII-induced contractions in rabbit aorta rings. The most active compounds had IC50 values in the nanomolar range. In conscious rats, compounds 4 and 21 antagonized the AII pressor response when administered orally. Compound 21 (SR 47436) was the most active; it was recently shown to also be active in cynomolgus monkeys both intravenously and orally. This molecule is now undergoing clinical trials for the treatment of hypertension. PMID- 8230128 TI - Synthesis, optical resolution, absolute configuration, and preliminary pharmacology of (+)- and (-)-cis-2,3,3a,4,5,9b-hexahydro-1-methyl-1H- pyrrolo [3,2-h]isoquinoline, a structural analog of nicotine. AB - Title compound, 8, has been synthesized from isoquinolinone, 1 (an improved preparation for which is presented) and separated into its antipodes with D- and L-di-p-toluoyltartaric acids. These antipodes and the racemic precursor have been evaluated (and found active) in two in vivo systems for their effects. The most potent of the three, (+)-8, has an ED50 of 7.13 mumol/kg for inhibition of spontaneous activity and 7.45 mumol/kg for antinociception compared to 4.44 and 4.81 mumol/kg, respectively, for (S)-(-)-nicotine. Compounds (-)-8 and 7 are about one-fourth as potent. Isomer (+)-8 has the 3aR,9bS configuration, the latter corresponding to (S)-(-)-nicotine as determined by X-ray crystallography. However, (+)-8 failed to compete for [3H]-nicotine binding, and its pharmacological effects were not blocked by mecamylamine. These bridged nicotine analogs either are binding to an as-yet-unidentified nicotinic receptor or they represent a novel class of non-nicotinic analgesics. PMID- 8230129 TI - 3-Acyl-4-hydroxyquinolin-2(1H)-ones. Systemically active anticonvulsants acting by antagonism at the glycine site of the N-methyl-D-aspartate receptor complex. AB - Most full antagonists at the glycine site of the NMDA receptor contain a carboxylic acid, which we believe to be detrimental to penetration of the blood brain barrier. By consideration of a pharmacophore, novel antagonists at this site have been designed in which the anionic functionality is a vinylogous acid, in the form of a 4-hydroxyquinolin-2(1H)-one. In this series, a 3-substituent is necessary for binding, and correct manipulation of this group leads to compounds such as the 3-(3-hydroxyphenyl)propargyl ester 24 (L-701,273), with an IC50 for displacement of [3H]-L-689,560 binding of 0.17 microM and Kb against NMDA in the cortical slice of 1.39 microM. Compounds were tested for their ability to prevent audiogenic seizure in DBA/2 mice; the most potent compound in this series is the cyclopropyl ketone 42 (L-701,252), with an ED50 of 4.1 mg/kg ip. A model is proposed for binding to the glycine site, in which an important interaction is of a putative receptor cation with the pi-system of the 3-substituent. PMID- 8230130 TI - 3-Nitro-3,4-dihydro-2(1H)-quinolones. Excitatory amino acid antagonists acting at glycine-site NMDA and (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors. AB - 3,4-Dihydro-2(1H)-quinolones, evolved from 2-carboxy-1,2,3,4,- tetrahydroquinolines and 3-carboxy-4-hydroxy-2(1H)-quinolones, have been synthesized and evaluated in vitro for antagonist activity at the glycine site on the NMDA receptor and for AMPA [(RS)-alpha-amino-3- hydroxy-5-methyl-4 isoxazolepropionic acid] antagonist activity. Generally poor potency at the glycine site is observed when a variety of electron-withdrawing substituents are attached to the 3-position of 3,4-dihydro-2(1H)-quinolones. The analogues 5-9 (IC50 values > 100 microM, Table I) exist largely in the 3,4-dipseudoaxial conformation (as evidenced by 1H NMR spectra), whereas the 3-cyano derivative (10, IC50 = 12.0 microM) has a relatively high population of the 3 pseudoequatorial conformer. The 3-nitro analogue (4, IC50 = 1.32 microM) has a pKa approximately 5 and thus exists at physiological pH as an anion with the nitro group planar to the quinolone ring. The general requirement of acidity for high affinity binding at the glycine/NMDA site is supported with the good activity of the other 3-nitro derivatives (13-21), all of which are deprotonated at physiological pH. The 3-nitro-3,4-dihydro-2(1H)-quinolones and 2-carboxy 1,2,3,4-tetrahydroquinolines show quite different structure-activity relationships at the 4-position. The unselective excitatory amino acid activity of 21 is comparable with 6,7-dichloro-quinoxaline-2,3-dione and 6,7 dichloroquinoxalic acid and this suggests similarities in their modes of binding to excitatory amino acid receptors. The broad spectrum excitatory amino acid antagonist activity of the 4-unsubstituted analogue 21 (KbNMDA = 6.7 microM, KbAMPA = 9.2 microM) and the glycine/NMDA selectivity of the other 3-nitro derivatives allows the proposal of a model for AMPA receptor binding which differs from the glycine binding pharmacophore in that there is bulk intolerance adjacent to the 4-position. Compound 21 (L-698,544) is active (ED50 = 13.2 mg/kg) in the DBA/2 mouse anticonvulsant model and is the most potent combined glycine/NMDA-AMPA antagonist yet reported, in vivo, and may prove to be a useful pharmacological tool. PMID- 8230131 TI - Orally active central dopamine and serotonin receptor ligands: 5-, 6-, 7-, and 8 [[trifluoromethyl)sulfonyl]oxy]-2-(di-n-propylamino)tetralins and the formation of active metabolites in vivo. AB - The racemic triflate derivatives 5-8 of the 5-, 6-, 7-, and 8-hydroxylated 2-(di n-propylamino)-teralins 1-4 were shown to possess similar pharmacological profiles to their phenolic counterparts in in vitro binding and in vivo biochemical and behavioral assays in rats. Consequently, subcutaneous administration of the 5-, 6-, and 7-triflates displayed essentially dopaminergic agonist properties, while the 8-triflate was shown to be a selective 5-HT1A receptor agonist. With respect to their agonist activities, the triflates were less potent than their phenolic analogs. The absolute oral bioavailability of compound 8 (8-triflate) was 4-5 times greater than the corresponding hydroxylated compound. Interestingly, in the in vivo biochemical assay compound 8 was found to be more potent after oral than after subcutaneous administration, indicating formation of one or more active metabolites. Following a study of the metabolism of compound 8 in rat hepatocytes, the monopropyl analog 9 was identified as the major metabolite and was surprisingly found to be more potent than compound 8. Oral administration of compound 5 (5-triflate) resulted in behavioral and biochemical effects indicative of mixed DA/5-HT1A agonist properties not seen after subcutaneous administration. These results may also be indicative of the formation of active metabolites. PMID- 8230132 TI - New adenosine kinase inhibitors with oral antiinflammatory activity: synthesis and biological evaluation. AB - Several 5-iodotubercidin analogues in the pyrazolo[3,4-d]pyrimidine ring system were synthesized as potential inhibitors of adenosine kinase by a direct Lewis acid-catalyzed glycosylation procedure using both the preformed carbohydrate and the heterocyclic base as starting materials. The 5'-hydroxyl, -chloro, -azido, deoxy, -amino, and -fluoro derivatives were prepared and evaluated in three systems for biological activity relative to adenosine, the true substrate, and 5 iodotubercidin, a known inhibitor. First, each compound was studied kinetically for inhibition of purified human placental adenosine kinase activity. The order of potency was: iodotubercidin > hydroxyl > amino > or = deoxy > fluoro > chloro >> azido. The Ki values for the 5'-hydroxyl and 5'-amino compounds, the two most potent inhibitors, were 80 and 150 nM, respectively. The inhibition appeared to be essentially competitive in nature, although a noncompetitive component of significance for the more potent inhibitors cannot be ruled out. Second, a bioassay was conducted in which the toxicity of 6-methylmercaptopurine riboside toward human CEM lymphoblasts was reversed by varying concentrations of the compounds. The order of effectiveness of the compounds in this system, representing a functional inhibition of adenosine kinase in cultured cells, was about the same as that with the purified enzyme, except that the 5'-chloro and 5' fluoro compounds were ineffective. Third, the 5'-hydroxyl derivative was evaluated in vivo in a rat pleurisy inflammation model and displayed biological activity at a dose of 30 mg/kg given orally. Finally, the in vitro toxicity of each compound was assessed in CEM lymphoblasts. Results indicated that the two most potent inhibitors in the pyrazolo[3,4-d]pyrimidine ring system, the 5' hydroxyl (7) and the 5'-amino (20), were 15-fold and 75-fold, respectively, less growth inhibitory than 5-iodotubercidin. PMID- 8230133 TI - Alkylpurines as immunopotentiating agents. Synthesis and antiviral activity of certain alkylguanines. AB - Several simple 8-substituted 9-alkyl- and 7,8-disubstituted 9-alkylguanine derivatives were synthesized as potential antiviral agents. These were tested for antiviral protection against a lethal Semliki Forest virus (SFV) infection in mice, and their antiviral properties were evaluated from a structure-activity standpoint. In this model system, 9-alkylguanines with the alkyl chain consisting of four to six carbons were found to be the most active. Substitution of the 8 position of the purine ring did not enhance activity, with the exception of the 7 alkyl-8-oxo substituent. These data were found to support the hypothesis that guanines need not contain an intact carbohydrate moiety in order to exhibit antiviral activity by virtue of immune potentiation. Hence, phosphorylation of guanosine analogs that exhibit antiviral activity by a similar mechanism does not play a significant role. PMID- 8230134 TI - Synthesis of 5-methyl-5-deaza nonclassical antifolates as inhibitors of dihydrofolate reductases and as potential antipneumocystis, antitoxoplasma, and antitumor agents. AB - A series of 2,4-diamino-5-methyl-6-(anilinomethyl)pyrido[2,3-d]pyrimidines 4-9 were synthesized as 5-deaza nonclassical antifolates containing trimethoxy, dichloro-, or trichlorophenyl substitutions and a N-H, N-CH3, or N-CHO at the 10 position. The compounds were evaluated as inhibitors of dihydrofolate reductases (DHFR) from Pneumocystis carinii (P. carinii), Toxoplasma gondii (T. gondii), rat liver (RL), and Lactobacillus casei (L. casei); as inhibitors of T. gondii and P. carinii cell growth in culture; and as antitumor agents. The compounds were prepared by modifications of procedures for classical 5-deaza folates. 2,4 Diamino-5-methyl-6-[(3',4',5'-trimethoxy-N- methylanilino)methyl]pyrido[2,3 d]pyrimidine (5a) exhibited high potency as well as selectivity (compared to RL DHFR) for P. carinii and T. gondii DHFR. Compound 5a is one of the most potent and selective nonclassical folate inhibitors of T. gondii DHFR known. The N-10 formyl analogue 2,4-diamino-5-methyl-6-[(N-formyl-3',4',5'-trimethoxyanilino) methyl]pyrido-[2,3-d]pyrimidine (6a) had decreased potency, but it maintained high selectivity for T. gondii DHFR. The corresponding chloro-substituted analogues maintained potency or had decreased potency; N-10 substitution did not increase potency or selectivity to the extent observed in the 3',4',5'-trimethoxy series. Partial reduction of the B ring to afford the dihydro analogue 2,4 diamino-5-methyl-6-[(N-formyl-3',4',5'-trimethoxyanilino) methyl]-5,8 dihydropyrido[2,3-d]pyrimidine (7), its 5,6,7,8-tetrahydropyrido[2,3-d]pyrimidine analogue 8, and 2,4-diamino-5-methyl-6-[(3',4',5'-trimethoxyanilino)methyl] 5,6,7, 8- tetrahydropyrido[2,3-d]pyrimidine (9) resulted in a significant decrease in potency. In T. gondii cell culture inhibitory studies, 2,4-diamino-5 methyl-6-[(3',4',5'- trimethoxyanilino)methyl]pyrido[2,3-d]pyrimidine (4a), 5a, and 6a were less potent compared to their DHFR inhibitory potencies. Against P. carinii cells in culture, 4a and 5a at 10 micrograms/mL were as effective as the clinically used combination of trimethoprim/sulfamethoxazole (50/250 micrograms/mL). With the exception of the B ring reduced analogues 7-9, all of the compounds were significantly cytotoxic to leukemia CCRF-CEM cells in culture. The chloro-substituted analogues, in general, were more potent against a variety of other tumor cells in culture than the trimethoxy analogues. These results were corroborated by the preclinical tumor screening program at the National Cancer Institute where the most potent compound 2,4-diamino-5-methyl-6-[(3',4' dichloroanilino)methyl]pyrido[2,3- d]pyrimidine (4b) was found to inhibit the growth of 26 tumor cell lines at an IG50 < 1.00 x 10(-8) M. PMID- 8230135 TI - Fluorocyclopropyl quinolones. 1. Synthesis and structure-activity relationships of 1-(2-fluorocyclopropyl)-3-pyridonecarboxylic acid antibacterial agents. AB - A series of 1-(2-fluorocyclopropyl)-3-pyridonecarboxylic acids has been prepared. These derivatives are characterized by having a fluorine atom at the 2-position on the cyclopropane ring as the N1 substituent and consist of both cis and trans stereoisomers. Structure-activity relationship studies indicate that the cis derivatives are more potent against Gram-positive bacteria than the corresponding trans counterparts, but the difference in potency against most Gram-negative bacteria is much smaller. The inhibitory effect of compounds 4, 5, 26, 27, 38, and 39 on supercoiling activity of DNA gyrase obtained from E. coli KL-16 correlated with their MICs against the same strain and also depend on their (26, 27, 38, 39) stereochemistry. Introduction of a fluorine atom on the cyclopropyl group resulted in the reduction of lipophilicity compared with the corresponding nonfluorinated quinolones. PMID- 8230136 TI - Quinolinecarboxylic acids. 3. Synthesis and antibacterial evaluation of 2 substituted-7-oxo-2,3-dihydro-7H-pyrido[1,2,3-de] [1,4]benzothiazine-6-carboxylic acids related to rufloxacin. AB - A series of 2-substituted-7-oxo-2,3-dihydro-7H-pyrido[1,2,3-de][1,4]benzothiazine 6 -carboxylic acids has been prepared and evaluated for in vitro antibacterial activity. These derivatives were less active than corresponding desmethylated analogues. Among these derivatives, the most active compound 22a was selected for preliminary pharmacokinetics in rats. The pharmacokinetic data indicated that 22a was rapidly absorbed and induced lasting plasma and urinary levels. In comparison with rufloxacin, it was excreted in low quantity in urine; a significant amount of desmethylated piperazinyl urinary metabolite was observed. PMID- 8230137 TI - 9-[(Phosphonoalkyl)benzyl]guanines. Multisubstrate analogue inhibitors of human erythrocyte purine nucleoside phosphorylase. AB - A series of 9-[(phosphonoalkyl)benzyl]guanines was synthesized and tested for inhibition of human erythrocyte purine nucleoside phosphorylase (PNPase). Inhibitors of PNPase should be T-cell selective, immunosuppressive agents with potential clinical utility in the treatment of a wide variety of disorders in which T-lymphocytes are pathogenic. An initial set of six analogues of the weak PNPase inhibitor 9-benzylguanine (2) contained a phosphonic acid group linked to the ortho, meta, or para position of the aryl moiety via two- or three-atom spacers. These compounds allowed us to probe for a favorable interaction with the phosphate-binding domain. Several additional meta phosphonoalkyl substituents were examined in an effort to optimize the spacer. The two most potent compounds, [[3-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl)methyl] benzyl]oxy] methylphosphonic acid (3f) and [[3-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9 methyl)methyl] benzyl]-thio]methylphosphonic acid (3j), were inhibitors of PNPase with Ki's of 5.8 and 1.1 nM, respectively. These inhibitors displayed competitive kinetics with respect to inosine and inorganic phosphate, which showed that these compounds possess binding determinants for both the purine- and phosphate-binding domains of the enzyme, characteristics that are consistent with 3f and 3j being multisubstrate analogue inhibitors of PNPase. The potency of 9-benzylguanine (2) was enhanced more than 6000-fold by linking a phosphonic acid residue with a (methylthio)methyl spacer to the meta position of 2 to give 3j, which illustrates the potent enzyme inhibitory properties available to multisubstrate analogue inhibitors. PMID- 8230138 TI - Benzoquinazoline inhibitors of thymidylate synthase: enzyme inhibitory activity and cytotoxicity of some sulfonamidobenzoylglutamate and related derivatives. AB - Several folate-like thymidylate synthase inhibitors are described in which the pteridine nucleus of the folic acid molecule is replaced by a benzoquinazoline moiety, which in turn is attached to the benzoylglutamate side chain by a sulfonamide link. The most potent compounds had Ki values as low as 2.5 nM against the human enzyme, were good substrates for the cellular reduced folate transport system and for folylpolyglutamate synthetase, and had IC50 values for growth inhibition of tumor cell lines as low as 70 nM. PMID- 8230139 TI - Peptide alpha-keto ester, alpha-keto amide, and alpha-keto acid inhibitors of calpains and other cysteine proteases. AB - A series of dipeptidyl and tripeptidyl alpha-keto esters, alpha-keto amides, and alpha-keto acids having leucine in the P2 position were synthesized and evaluated as inhibitors for the cysteine proteases calpain I, calpain II, cathepsin B, and papain. In general, peptidyl alpha-keto acids were more inhibitory toward calpain I and II than alpha-keto amides, which in turn were more effective than alpha keto esters. In the series Z-Leu-AA-COOEt, the inhibitory potency decreased in the order: Met (lowest KI) > Nva > Phe > 4-Cl-Phe > Abu > Nle (highest KI) with calpain I, while almost the reverse order was observed for calpain II. Extending the dipeptide alpha-keto ester to a tripeptide alpha-keto ester yielded significant enhancement in the inhibitory potency toward cathepsin B, but smaller changes toward the calpains. Changing the ester group in the alpha-keto esters did not substantially decrease KI values for calpain I and calpain II. N Monosubstituted alpha-keto amides were better inhibitors than the corresponding alpha-keto esters. alpha-Keto amides with hydrophobic alkyl groups or alkyl groups with an attached phenyl group had the lower KI values. N,N-Disubstituted alpha-keto amides were much less potent inhibitors than the corresponding N monosubstituted peptide alpha-keto amides. The peptide alpha-keto acid Z-Leu-Phe COOH was the best inhibitor for calpain I (KI = 0.0085 microM) and calpain II (KI = 0.0057 microM) discovered in this study. It is likely that the inhibitors are transition-state analogs and form tetrahedral adducts with the active site cysteine of cysteine proteases and form hydrogen bonds with the active site histidine and possibly another hydrogen bond donor in the case of monosubstituted amides. Several inhibitors prevented spectrin degradation in a platelet membrane permeability assay and may be useful for the treatment of diseases which involve neurodegeneration. PMID- 8230140 TI - A tight binding inhibitor of 5-aminoimidazole ribonucleotide carboxylase. PMID- 8230141 TI - Is there an important moral distinction for medical ethics between lying and other forms of deception? PMID- 8230142 TI - Medical involvement in torture. PMID- 8230143 TI - The ethics of medical involvement in torture. AB - The difficulties of establishing a definition of torture are discussed, and a definition is suggested. It is then argued that, irrespective of general ethical questions, doctors in particular should never be involved because of their social role. PMID- 8230144 TI - The ethics of medical involvement in torture: commentary. PMID- 8230145 TI - Maturity of children to consent to medical research: the babysitter test. AB - The age of maturity of children to consent for medical research is under debate, as different authorities regard the capacity of young teenagers as either satisfactory or not to grant consent without parental participation in the process. The present paper contrasts the generally accepted guideline for ethics in paediatric research in Canada with what the same children are allowed and expected to be able to do as babysitters. This comparison reveals deep incongruences in the way the maturity of the same children is appreciated for two different tasks. PMID- 8230146 TI - Attitudes of healthcare professionals and the public towards the sale of kidneys for transplantation. AB - We conducted a survey of attitudes towards the sale of kidneys for transplantation within and without the medical community. Half of those polled received a case of a young man in India whose only chance for survival was to purchase a kidney, the other half a case of a Canadian man who was suffering side effects from dialysis and had been on the transplant waiting list for three years. We found the percentage of responses allowing the patients to purchase a kidney was similar for the two cases (40 per cent in the Canadian case and 49 per cent in the Indian case). The medical groups had much lower rates of approval of this practice than the public. In all groups those allowing the practice showed similar concerns about regulation. This survey indicates public opinion to be contrary to public policy. PMID- 8230147 TI - The Hippocratic contract. PMID- 8230148 TI - A randomized trial of ethics education for medical house officers. AB - We report the results of a randomized trial to assess the impact of an innovative ethics curriculum on the knowledge and confidence of 85 medical house officers in a university hospital programme, as well as their responses to a simulated clinical case. Twenty-five per cent of the house officers received a lecture series (Limited Intervention or LI), 25 per cent received lectures and case conferences, with an ethicist in attendance (Extensive Intervention or EI), and 50 per cent served as controls. A post-intervention questionnaire was administered. Knowledge scores did not differ among the groups. Confidence regarding ethical issues was significantly greater in the aggregate intervention group (3.9 on a 1 to 5 scale) compared to the control group (3.6). Confidence regarding procedural issues related to ethics was significantly higher for the EI group than for the controls (4.0 v 2.8). Responses to a simulated case showed that significantly fewer house officers in the EI group would intubate a patient for whom such therapy would be futile (EI = 57 per cent, LI = 87 per cent, Controls = 82 per cent). We conclude that ethics education can have an impact on house officers' confidence and their responses to a simulated case, and that the EI was more effective than the LI. Such results have implications regarding the implementation of ethics education during residency. PMID- 8230149 TI - The ethics of postmortem examinations in contemporary Islam. AB - Postmortem examinations have recently become common practice in Western medicine: they are used to verify the cause of death and to obtain additional scientific information on certain diseases, as well as to train medical students. For religious people of the monotheistic faiths postmortems present several ethical questions even though the advantages attributed to postmortems in the West are also acknowledged by Jews, Christians and Muslims. The Islamic way of dealing with such questions will be surveyed via contemporary fatawa (legal opinions) issued primarily by Egyptian scholars; Islamic law, which was formulated in the eighth to ninth centuries, did not speak of postmortems. I will therefore depict the means whereby contemporary scholars approach postmortems in the absence of clear legal reference. The difficulties that postmortems create for Muslims at present will be weighed against some shar i instructions which may help circumvent them. While the ethical and religious debate continues, postmortems seem to be accepted but not, however, without certain reservations. PMID- 8230150 TI - The great slippery-slope argument. AB - Whenever some form of beneficent killing--for example, voluntary euthanasia--is advocated, the proposal is greeted with a flood of slippery-slope arguments warning of the dangers of a Nazi-style slide into genocide. This paper is an attempt systematically to evaluate arguments of this kind. Although there are slippery-slope arguments that are sound and convincing, typical formulations of the Nazi-invoking argument are found to be seriously deficient both in logical rigour and in the social history and psychology required as a scholarly underpinning. As an antidote, an attempt is made both to identify some of the likely causes of genocide and to isolate some of the more modest but legitimate fears that lie behind slippery-slope arguments of this kind. PMID- 8230151 TI - The ethics of experimental heroin maintenance. AB - In response to widespread concern about illegal drug use and the associated risk of the spread of HIV/AIDS, a study was undertaken to examine whether it was, in principle, feasible to conduct a trial providing heroin to dependent users in a controlled manner. Such a trial involves real ethical issues which are examined in this paper. The general issues examined are: should a trial be an experiment or an exercise in public policy?; acts and omissions; countermobilization; termination of a trial, and payment for drugs and for a trial. The specific issues examined are: selection of trial participants; privacy; issues for staff working on a trial; coupling the trial with other treatment, and issues for researchers. A number of alternative approaches to the various ethical issues are presented and discussed. PMID- 8230152 TI - On the morality of deception--does method matter? A reply to David Bakhurst. AB - Does it signify morally whether a deception is achieved by a lie or some other way? David Bakhurst has challenged my view that it can signify. Here I counter his criticisms--firstly, by clarifying the terminology: What counts as a lie? Secondly, by exploring further what makes lying wrong. Bakhurst maintains that lying is wrong in that it infringes autonomy--and other deceiving stratagems, he says, do so equally. I maintain that lying is wrong in that it endangers trust- and other types of deceiving stratagems do not do so equally. Lying to patients, I contend, is an abuse of their trust. Other forms of their intentional deception need not be so, although, in our autonomy-minded culture, they are likely to be so. PMID- 8230153 TI - CATCH 22. PMID- 8230154 TI - DiGeorge syndrome: an historical review of clinical and cytogenetic features. PMID- 8230155 TI - Prevalence of 22q11 microdeletions in DiGeorge and velocardiofacial syndromes: implications for genetic counselling and prenatal diagnosis. AB - Deletions of chromosome 22q11 have been seen in association with DiGeorge syndrome (DGS) and velocardiofacial syndrome (VCFS). In the present study, we analysed samples from 76 patients referred with a diagnosis of either DGS or VCFS to determine the prevalence of 22q11 deletions in these disorders. Using probes and cosmids from the DiGeorge critical region (DGCR), deletions of 22q11 were detected in 83% of DGS and 68% of VCFS patients by DNA dosage analysis, fluorescence in situ hybridisation, or by both methods. Combined with our previously reported patients, deletions have been detected in 88% of DGS and 76% of VCFS patients. The results of prenatal testing for 22q11 deletions by FISH in two pregnancies are presented. We conclude that FISH is an efficient and direct method for the detection of 22q11 deletions in subjects with features of DGS and VCFS as well as in pregnancies at high risk for a deletion. PMID- 8230156 TI - Isolation of a new marker and conserved sequences close to the DiGeorge syndrome marker HP500 (D22S134). AB - End fragment cloning from a YAC at the D22S134 locus allowed the isolation of a new probe HD7k. This marker detects hemizygosity in two patients previously shown to be dizygous for D22S134. This positions the distal deletion breakpoint in these patients to the sequences within the YAC, and confirms that HD7k is proximal to D22S134. In a search for coding sequences within the region commonly deleted in DGS we have identified a conserved sequence at D22S134. Although no cDNAs have yet been isolated, genomic sequencing shows a short open reading frame with weak similarity to collagen proteins. PMID- 8230157 TI - Conotruncal anomaly face syndrome is associated with a deletion within chromosome 22q11. AB - The conotruncal anomaly face syndrome was described in a Japanese publication in 1976 and comprises dysmorphic facial appearance and outflow tract defects of the heart. The authors subsequently noted similarities to Shprintzen syndrome and DiGeorge syndrome. Chromosome analysis in five cases did not show a deletion at high resolution, but fluorescent in situ hybridisation using probe DO832 showed a deletion within chromosome 22q11 in all cases. PMID- 8230158 TI - Velocardiofacial syndrome in a mother and daughter: variability of the clinical phenotype. AB - We report a mother and daughter with features of the velocardiofacial (VCF) syndrome and monosomy for 22q11 identified using molecular techniques. The mother had surgery as a child for a cleft palate and a congenital heart defect, and her facial features were consistent with the diagnosis. The daughter had developmental delay, absent speech, scoliosis, and similar facial features, but no cleft palate or congenital heart defect. These cases illustrate the considerable intrafamilial variability of the phenotype of VCF syndrome. The clinical and molecular diagnosis of this syndrome is discussed. The phenotypic variability of the VCF syndrome means that many cases may be undiagnosed. PMID- 8230159 TI - High resolution mapping of interstitial long arm deletions of chromosome 16: relationship to phenotype. AB - The breakpoints of seven interstitial deletions of the long arm of chromosome 16 and two ring chromosomes of this chromosome were mapped by in situ hybridisation or by analysis of mouse/human somatic cell hybrids containing the deleted chromosome 16. Use of a high resolution cytogenetic based physical map of chromosome 16 enabled breakpoints to be assigned to an average resolution of at least 1.6 Mb. In general, interstitial deletions involving q12 or q22.1 have broadly similar phenotypes though there are differences in specific abnormalities. Deletions involving regions more distal, from 16q22.1 to 16q24.1, were associated with relatively mild dysmorphism. One region of the long arm, q24.2 to q24.3, was not involved in any deletion, either in this study or in any previous report. Presumably, monosomy for this region is lethal. In contrast, patients with deletions of 16q21 have a normal phenotype. These results are consistent with the proposed distribution of genes, frequent in telomeric Giesma light band regions but infrequent in G positive bands. PMID- 8230160 TI - Analysis of a terminal Xp22.3 deletion in a patient with six monogenic disorders: implications for the mapping of X linked ocular albinism. AB - The molecular characterisation of chromosomal aberrations in Xp22.3 has established the map position of several genes with mutations resulting in diverse phenotypes such as short stature (SS), chondrodysplasia punctata (CDPX), mental retardation (MRX), ichthyosis (XLI), and Kallmann syndrome (KAL). We describe the clinical symptoms of a patient with a complex syndrome compatible with all these conditions plus ocular albinism (OA1). He has a terminal Xp deletion of at least 10 Mb of DNA. Both the mother and sister of the patient are carriers of the deletion and show a number of traits seen in Turner's syndrome. The diagnosis of ocular albinism was confirmed in the patient and his mother, who shows iris translucency, patches and streaks of hypopigmentation in the fundus, and macromelanosomes in epidermal melanocytes. By comparative deletion mapping we can define a deletion interval, which locates the OA1 gene proximal to DXS143 and distal to DXS85, with the breakpoints providing valuable starting points for cloning strategies. PMID- 8230161 TI - Upper and lower neural tube defects: an alternate hypothesis. AB - It has been suggested that neural tube defects (NTDs) of the upper type (anencephaly, encephalocele, and thoracic spina bifida) may have a pathogenesis different from those of the lower type (lumbosacral spina bifida), since recurrent cases within a sibship were said always to be concordant with respect to NTD type. Also, spontaneous abortion, additional malformation, and recurrence rate were observed to be higher in the upper group, and there was an excess of females in upper NTD probands. To test this hypothesis, we measured the above variables in upper and lower NTDs in a sample from Quebec. We found less than full concordance (50%) of NTD type in 18 sib pairs. Recurrence rate was not significantly lower in the lower NTD group (5.6 v 5.8%). The other variables were in general agreement with previous studies, inconsistent findings possibly attributable to different NTD population incidences. These findings can be accounted for if upper and lower NTDs share a similar pathogenesis and the embryo is more susceptible during early than late neural tube formation. PMID- 8230162 TI - DiGeorge syndrome: part of CATCH 22. AB - DiGeorge syndrome (DGS) comprises thymic hypoplasia, hypocalcaemia, outflow tract defects of the heart, and dysmorphic facies. It results in almost all cases from a deletion within chromosome 22q11. We report the clinical findings in 44 cases. We propose that DiGeorge syndrome should be seen as the severe end of the clinical spectrum embraced by the acronym CATCH 22 syndrome; Cardiac defects, Abnormal facies, Thymic hypoplasia, Cleft palate, and Hypocalcaemia resulting from 22q11 deletions. PMID- 8230163 TI - Disease gene mapping in isolated human populations: the example of Finland. PMID- 8230164 TI - Mapping of a gene for non-specific X linked mental retardation: evidence for linkage to chromosomal region Xp21.1-Xp22.3. AB - Linkage analysis of a non-specific form of X linked mental retardation (MRX) was performed with 16 polymorphic markers spanning the entire X chromosome in a three generation Italian family, including four male patients with moderate mental retardation. One obligate carrier woman had mild mental retardation and another two had normal intelligence. The results indicate tight linkage to DNA markers DXS84 (L754), DXS164 (pERT87-15), and DXS278 (CRI-S232). A maximum lod score of 2.11 at theta = 0.00 was obtained with DXS164 and DXS278. The linked region spanned chromosomal bands Xp21.1-Xp22.3, that is, the same portion of the X chromosome where MRX2 and MRX10-13 have been previously localised. PMID- 8230165 TI - Orofaciodigital syndrome type III in two sibs. AB - A brother and sister with mental retardation, malformations of the cerebellar vermis, characteristic 'metronome' eye movements, lingual hamartomas, and postaxial polydactyly are described. The clinical overlap with the OFD group of syndromes, in particular OFD III and OFD VI, and Joubert's syndrome is discussed. If OFD III syndrome is to be recognised as a distinct entity, these two cases represent the second and third cases reported. PMID- 8230166 TI - Sex linked valvular dysplasia. AB - A family is described in which three males have been affected by congenital valvular dysplasia of one or more heart valves, in one case leading to neonatal death. The pedigree is consistent with sex linked inheritance. PMID- 8230167 TI - Airway abnormalities in Jarcho-Levin syndrome: a report of two cases. AB - Two infants with the Jarcho-Levin syndrome of vertebral anomalies underwent flexible fibre optic bronchoscopy. Central airway abnormalities not amenable to surgical correction were found in both patients. These abnormalities may contribute significantly to the respiratory failure seen in affected infants, and should be considered when evaluating continuing medical support. PMID- 8230168 TI - Long survival of a patient with Marshall-Smith syndrome without respiratory complications. AB - The Marshall-Smith syndrome is characterised by overgrowth, accelerated skeletal maturation, and dysmorphic facial features, often associated with mental retardation of variable degree. Most of the reported patients died in the first three years of life mainly because of respiratory problems. We describe a 5 year old patient with this rare syndrome, who has optic atrophy and agenesis of the corpus callosum, but has no respiratory problems so far. This observation underlines the clinical variability of the Marshall-Smith syndrome and indicates that life expectancy may be prolonged. PMID- 8230169 TI - Prenatal diagnosis of a giant intracranial teratoma associated with pulmonary hypoplasia. AB - We present a case in which an intracranial tumour replacing all intracranial contents was diagnosed by sonography at 31 weeks' gestation. The patient was delivered by caesarean section and died shortly after delivery. At necropsy, the tumour was found to be a teratoma with no recognisable normal brain tissue present. Additional findings at necropsy included pulmonary and adrenal hypoplasia. The diagnosis and prognosis of intracranial teratomas diagnosed in utero, and the association of this tumour with pulmonary hypoplasia, are discussed. PMID- 8230170 TI - Severe developmental delay and multiple strawberry naevi: a new syndrome? AB - An 18 month old girl with dysmorphic features, severe developmental delay, multiple strawberry naevi, and capillary naevi is described. No previous report of a similar association of features has been identified. PMID- 8230171 TI - Chromosome abnormalities and Williams-Beuren syndrome. PMID- 8230172 TI - Counselling pitfalls in Romano-Ward syndrome. PMID- 8230173 TI - Hepatocellular carcinoma associated with chronic Schistosoma mansoni infection in a chimpanzee. AB - Hepatocellular carcinoma associated with chronic Schistosoma mansoni infection in a chimpanzee, estimated to be a 12-year-old and born in West Africa, is reported. The hepatic tumor appeared as a solitary firm nodule, and histological examination revealed well-differentiated hepatocellular carcinoma with a trabecular pattern. Hepatitis B virus and hepatitis C virus infections were excluded by serological testing in that animal. This is the first report of hepatocellular carcinoma in the chimpanzee with schistosomiasis. PMID- 8230174 TI - Seroepidemiology of dengue and other arboviruses in a natural population of toque macaques (Macaca sinica) at Polonnaruwa, Sri Lanka. AB - A seroepidemiological study of arboviruses infecting 115 wild toque macaques (Macaca sinica) at Polonnaruwa, Sri Lanka showed a high prevalence of antibodies to dengue and Lumbo viruses. There was low seroprevalence of Chandipura (2/115) and Batai (1/115) virus antibodies, but no seropositivity to Chikungunya or Sindbis. There was no serological evidence of infection by Japanese encephalitis (JE) virus in spite of large human epidemics in the study area, indicating that toque macaques are unlikely to be an epidemiologically relevant host in the maintenance cycle of JE virus. PMID- 8230175 TI - Parathyroid hormone, ionised calcium, and potentially interacting variables in plasma of an Old World primate. AB - Bone turnover and calcium homeostasis in man can only be modelled validly in Old World nonhuman primates. In order to interpret the models it is necessary to establish endocrine and biochemical parameters of bone mineral metabolism. This report is probably the first description of acute phase parathyroid responses to manipulations of blood ionised calcium, and of reference values for potentially interacting variables, in vervet monkeys. Plasma parathyroid hormone concentrations were measured in vervets under defined conditions, and ranges reported as normal for other nonhuman primates and man are summarised. PMID- 8230176 TI - Multifactorial etiology of anemia in SIV-infected rhesus macaques: decreased BFU E formation, serologic evidence of autoimmune hemolysis, and an exuberant erythropoietin response. AB - We attempted to define the etiology of anemia in SIV-infected rhesus macaques. Bone marrow culture showed significantly decreased (75% reduction) burst forming unit-erythroid (BFU-E) growth in end-stage SIV+ "sick" animals. Direct antiglobulin tests (DAT) were positive in nine of 35 SIV+ "well" and 14 of 14 SIV+ "sick" monkeys (0 of 25 control animals had positive DATs). In animals with a positive DAT, moderate to severe anemia was observed, as was increased LDH and spherocytosis. Erythropoietin was measured in four control, eight SIV+ "well" and five SIV+ "sick" animals with mean levels of 4.0, 15.4, and 1176 mU/mL (r = .94) in the three groups. These data suggest that the cause of anemia in the SIV infected rhesus macaque is multifactorial, that there may be a defect in erythropoiesis, and that, serologically, an IgG mediated autoimmune hemolytic anemia is also present. PMID- 8230177 TI - Chronic Pseudomonas aeruginosa endobronchitis in rhesus monkeys: II. A histopathologic analysis. AB - We have recently established a rhesus monkey model of chronic Pseudomonas aeruginosa (PA) endobronchitis by bronchoscopic instillation of PA-embedded agar beads. All experimental animals developed chronic neutrophilic endobronchitis similar to chronic PA endobronchitis in cystic fibrosis (CF). Histopathologic studies further confirmed similarities to chronic PA endobronchitis in CF, including marked peribronchial inflammation, epithelial damage, presence of degraded cilia and ciliary abnormalities, appearance of PA bacterial clusters, mucosal hyperplasia, goblet cell hypertrophy/hypersecretion, airway obstruction, alveolar abnormalities, bronchiectasis, and fibrosis. PMID- 8230178 TI - Tubal ligation in the chimpanzee (Pan troglodytes) as a means of contraception. AB - Contraception of two chimpanzees was considered necessary for clinical reasons. After failure of reversible methods of contraception (an intrauterine device and a long-acting progestagen), tubal ligation was successfully performed, using a technique similar to one used in women. PMID- 8230179 TI - Stillbirth of twins in a squirrel monkey (Saimiri boliviensis peruviensis). AB - This is the first published report of twinning in a squirrel monkey (genus Saimiri). The mother survived but the twins, both male and close to full term, were stillborn. PMID- 8230180 TI - Acute gastric dilatation in two black and white colobus monkeys. AB - Acute gastric dilatation in two black and white colobus monkeys is described. The subfamily colobini have a ruminant-like stomach and a pregastric fermentation, of important for the observations described. PMID- 8230181 TI - Empirical use of metronidazole for treatment of diarrhea in laboratory rhesus macaques: an evaluation. PMID- 8230182 TI - Total elbow arthroplasty: salvage of unsuccessful previous elbow operations. AB - Six consecutive patients with a stiff, painful, nonfunctional elbow were reconstructed utilizing a semicontrained total elbow prosthetic replacement. All patients had multiple previous surgical procedures with unsuccessful results. The average preoperative flexion (80 degrees) and extension (-60 degrees) improved to 130 degrees to -10 degrees respectively. All patients were able to regain satisfactory use of the elbow without pain. PMID- 8230184 TI - The law regarding utilization review. PMID- 8230183 TI - Testicular vein and peripheral vein testosterone, follicle stimulating hormone, luteinizing hormone, and prolactin concentrations in a patient with androgen insensitivity syndrome. AB - Androgen insensitivity syndrome (testicular feminization) is classically diagnosed by history, physical examination, karyotype and ultimately exploratory laparotomy. In this case, the diagnosis was verified by amenorrhea, female phenotype, Barr body negative buccal smear, and finally, exploratory laparotomy which revealed bilateral abdominal testes. Testicular vein blood samples obtained at the time of surgery were analyzed for testosterone, follicle stimulating hormone, luteinizing hormone, and prolactin and compared to peripheral vein blood samples. The testosterone concentration in the testicular vein was twice the concentration in the peripheral sample, reflecting gonadal function. The other testicular vein hormone concentrations mirrored the peripheral vein concentrations. PMID- 8230185 TI - Affordable health care. PMID- 8230186 TI - The University of Mississippi Medical Center's Antenatal Diagnostic Unit: on the cutting edge of tomorrow. AB - Prenatal diagnosis is a rapidly expanding field in the specialty of obstetrics. Modern providers of perinatal care must understand the fundamentals of inherited disease and keep abreast of advances in antenatal diagnosis. Pivotal in providing appropriate evaluation in this area is a referral center that is capable of affording the necessary evaluation, documentation and consultation. Although pregnancy termination is an option for some couples, many will use the information provided to prepare emotionally and financially for the birth of a child with special problems and needs. Some diagnoses will change obstetric management during the remainder of pregnancy or will lead to a recommendation for delivery at a tertiary medical facility. Other women with medical conditions warrant preconception counseling regarding pregnancy management and likelihood of a successful outcome. Patients with genetic disorders within their families may desire carrier detection through DNA analysis. Finally, women on medications at or near the time of conception frequently have concerns regarding potential teratogenesis. The ADU represents a commitment by both the Department of Obstetrics and Gynecology and the University of Mississippi Medical Center to furnish these services to providers and patients of the State of Mississippi. PMID- 8230187 TI - Women in medicine. PMID- 8230188 TI - Hippocrates revisited. PMID- 8230189 TI - A doctor's adventures in motherland. PMID- 8230190 TI - DNA deletion of mitochondrial introns is correlated with the process of senescence in Podospora anserina. AB - In the filamentous fungus Podospora anserina, the unavoidable phenomenon of senescence is associated with specific mitochondrial rearrangements and particularly with the amplification of some regions of the mitochondrial chromosome. Mechanisms responsible for these rearrangements are still unknown. The implication in this phenomenon, of the first intron of the mitochondrial gene cox1 (intron alpha), a class II intron that presents significant amino acid similarity with retroviral reverse transcriptases, was postulated several years ago. We demonstrate here by polymerase chain reaction experiments: (1) that senescent and young cultures contain DNA molecules precisely deleted for intronic sequences; (2) that these deletions are found to a much greater extent in senescent than in young cultures; (3) that DNA intron deletion likely results from a reverse transcriptase-mediated mechanism as indicated by the detection of copies of the gene 1 cox1 completely devoid of its 15 introns; (4) that the intron alpha-encoded protein could intervene in this process. On the whole, these results strongly suggest that in Podospora, an increase in a mitochondrial reverse transcriptase activity probably mediated by the intron alpha-encoded protein is involved in the process of senescence. PMID- 8230191 TI - Turnover of fluorescent nucleoside triphosphates by isolated immobilized myosin filaments. Transient kinetics on the zeptomole scale. AB - Recent developments of in vitro motility assays have allowed the sliding velocity and force generation to be measured when a single actin filament interacts with a small number of immobilized myosin molecules. In contrast, the associated ATPase activities have been estimated from the whole population of molecules in the flow cell using steady-state kinetics. For a more unambiguous estimate of the crossbridge step size, it would be desirable to measure the ATPase activity of those molecules actually under observation in the in vitro assay in real time. As a start to solving this formidable problem we have investigated the use of fluorescent ATP analogues as probes to measure the ATPase activity of immobilized myosin filaments. Turnover rates for the substrate analogue, FEDA-ATP (an analogue in which a fluorescein moiety is linked via an ethylenediamine chain to the ribose of ATP) were recorded by displacement of the steady-state intermediate with excess ATP. Using epifluorescence light microscopy, small clumps of rabbit skeletal and scallop striated muscle synthetic thick filaments and single native clam (Mercenaria) red adductor muscle thick filaments were assayed. The latter filaments contain several thousand myosin molecules and thus they represent the application of transient kinetic methodology on the zeptomole scale. In the presence of Ca2+, the derived rate constants for FEDA-ATP turnover are close to those expected for the same preparations in solution (0.06 s-1 for rabbit skeletal, 0.2 s-1 for scallop and clam adductor muscle myosins), indicating that immobilization does not have a significant effect on the ATPase activity. In the absence of Ca2+, molluscan preparations show a slower FEDA-ATP turnover rate but they do not appear as well regulated as in solution. In such microscope assays the observed displacement rate does not reflect the true turnover rate owing to photobleaching, and possibly regulatory light chain depletion. Future developments for extending this assay to the actin-activated state are discussed. PMID- 8230192 TI - A programmed translational frameshift is required for the synthesis of a bacteriophage lambda tail assembly protein. AB - Two proteins, one of 31 kDa and one of 16 kDa, are encoded by a segment of the phage lambda tail gene region that contains two overlapping reading frames, neither of which is long enough to encode the larger protein. We show that the abundant 16-kDa protein (gpG) is encoded by the upstream open reading frame, gene G. The 31-kDa protein, gpG-T, is encoded jointly by gene G and the overlapping downstream T open reading frame. gpG-T is synthesized as the result of a translational frameshift that occurs when a ribosome translating the G gene slips back by one nucleotide at a position six codons from the C terminus of the gene and thereby bypasses the G termination codon to continue on in the T open reading frame. The resulting protein shares 135 residues of N-terminal amino acid sequence with gpG, followed by 144 amino acid residues of unique sequence. The frameshift event occurs with a frequency of approximately 4% at the sequence G GGA AAG, which encodes the dipeptide -Gly-Lys- in both the zero and -1 reading frames. The frameshift frequencies of point mutants in this "slippery sequence" argue that codon-anticodon interactions with both the glycyl and the lysyl-tRNA are important for frameshifting to occur. We find no clear evidence for a pausing mechanism to enhance frameshifting, as is seen in other well-characterized frameshifts. No simple secondary structure has been predicted for the region downstream from the slippery sequence, but this downstream sequence does contribute to the frameshifting rate. Our results together with those of Katsura and Kuhl show that the frameshift product, gpG-T, has an essential role in lambda tail assembly, acting prior to tail shaft assembly. The role of gpG in tail assembly is not known. We find that both gpG and the gpG-T are absent from mature virions. PMID- 8230193 TI - Translation initiation complex formation with 30 S ribosomal particles mutated at conserved positions in the 3'-minor domain of 16 S RNA. AB - Escherichia coli 30 S ribosomal subunits containing in vitro (phage T7 RNA polymerase-generated) 16 S rRNA, both wild-type and mutant, were examined by toeprinting. These synthetic particles were used to compare the effects of the absence of base modification and of specific nucleotide substitutions in conserved sequence regions of the RNA on the assembly of mRNA, tRNAs and 30 S particles into a translational initiation complex. Initiation factor-3-dependent selection of tRNA(fMet) from a mixture of tRNA(fMet) and tRNA(Phe) occurred with all particles, although 20 times less initiation factor-3 was needed for the synthetic particles, including the mutants. Whereas isolated 30 S particles or those reconstituted with isolated RNA did not distinguish between tRNA(fMet) and tRNA(Phe) for ternary complex formation in the absence of initiation factor-3 (intrinsic selection ability), the synthetic particles preferred tRNA(fMet). The difference between the natural and synthetic particles appears to be due to the absence of certain base modifications, but not m2(6)A, in the synthetic RNA. Synthetic particles containing the mutation U1512C, which converts the universal U.G pair to C.G enhanced both tRNA(fMet) binding and selectivity, although other mutations at that site, namely U1512G, G1523A and U1512C/C1524U, had no such effect. Mutants U1498G and G1401C/C1501G, both located in a highly conserved single-stranded region of the 3'-minor domain, also enhanced tRNA(fMet) selectivity, in this case by reducing complex formation with elongator tRNA. Complex formation between elongator tRNA and the G1401C/C1501G mutant was reduced to almost undetectable levels. The results also indicated that the association rate for initiation complex formation for G1401C/C1501G was considerably lower than for the wild-type sequence. This result had not been detected by standard tRNA-30 S binding assays. Overall, the data suggest that (some of) the 16 S rRNA base modifications as well as the tertiary structure around the decoding site act to desensitize the intrinsic selection ability of the ribosome for tRNA(fMet). PMID- 8230194 TI - High-resolution crystal structures of distal histidine mutants of sperm whale myoglobin. AB - The highly conserved distal histidine residue (His64) of sperm whale myoglobin modulates the affinity of ligands. In an effort to fully characterize the effects of mutating residue 64, we have determined the high-resolution crystal structures of the Gly64, Val64, Leu64, Thr64 and Gln64 mutants in several liganded forms. Metmyoglobins with hydrophobic substitutions at residue 64 (Val64 and Leu64) lack a water molecule at the sixth coordination position, while those with polar amino acid residues at this position (wild-type and Gln64) retain a covalently bound water molecule. In the Thr64 mutant, the bound water position is only partially occupied. In contrast, mutating the distal histidine residue to glycine does not cause loss of the coordinated water molecule, because the hydrogen bond from the imidazole side-chain is replaced by one from a well-ordered solvent water molecule. Differences in water structure around the distal pocket are apparent also in the structures of deoxymyoglobin mutants. The water molecule that is hydrogen-bonded to the N epsilon atom of histidine 64 in wild-type deoxymyoglobin is not found in any of the position 64 mutant structures that were determined. Comparison of the carbonmonoxy structures of wild-type, Gly64, Leu64 and Gln64 myoglobins in the P6 crystal form shows that the conformation of the Fe-C-O complex is nearly linear and is independent of the identity of the amino acid residue at position 64. However, the effect of CO binding on the conformation of residue 64 is striking. Superposition of deoxy and carbonmonoxy structures reveals significant displacements of the residue 64 side-chain in the wild-type and Gln64 myoglobins, but no displacement in the Leu64 mutant. These detailed structural studies provide key insights into the mechanisms of ligand binding and discrimination in myoglobin. PMID- 8230195 TI - Orthorhombic crystal form of bacteriorhodopsin nucleated on benzamidine diffracting to 3.6 A resolution. AB - Freshly formed benzamidine crystals were found to provide a suitable nucleation surface for crystallisation of bacteriorhodopsin. At 20 degrees C and 1% octylglucoside pseudohexagonal needles of bacteriorhodopsin nucleated on the benzamidine surface. At 4 degrees C and reduced detergent concentration (0.5%) a new, better-ordered orthorhombic crystal form of bacteriorhodopsin was formed by heterogeneous nucleation on benzamidine. Polarised absorption spectroscopy and flash photolysis experiments were used to show that the crystalline bacteriorhodopsin is photoactive in both forms. The M-intermediate absorbs maximally at 405nm and formation of M does not disturb the crystal lattice. The plate-shaped crystals diffract to a resolution of 3.6 A along the a and b directions and to 4.2 A in the c direction. The most likely space group of the crystals is C222 (a = 107.5 A, b = 117.0 A, c = 69.5 A). The crystals are built from layers of bacteriorhodopsin molecules that are tilted from the c axis by about 45 degrees. In addition, as a background to the discovery of the new crystal form, the influence of different detergents, additives and precipitants on the formation of pseudohexagonal needles is presented. PMID- 8230196 TI - Structure of a phosphonate-inhibited beta-lactamase. An analog of the tetrahedral transition state/intermediate of beta-lactam hydrolysis. AB - The crystal structure of beta-lactamase from Staphylococcus aureus inactivated by p-nitrophenyl[[N-(benzyloxycarbonyl)amino]methyl]phosphonate, a methylphosphonate monoester monoanion inhibitor, has been determined and refined at 2.3 A resolution. The structure reveals a tetrahedral phosphorus covalently bonded to the O gamma atom of the active site serine, Ser70. One of the oxygen atoms bonded to phosphorus is located in the oxyanion hole formed by the two main-chain nitrogen atoms of Ser70 and Gln237, and the second bonded oxygen is solvated. The (benzyloxycarbonyl)aminomethyl group is oriented towards the active site gully such that the peptide group forms compensating electrostatic interactions with polar groups on the enzyme. The benzyl group forms a hydrophobic interaction with Ile239 and an aromatic-aromatic edge-to-face interaction with Tyr105, which has undergone a conformational transition relative to the native structure. The mode of binding supports the proposal that on reaction with the enzyme, the phosphonate generates a structure analogous to the tetrahedral transition state/intermediate associated with the acylation step of a normal substrate. The disposition of the phosphonyl group in this complex is the same as that of the corresponding phosphoryl group in the complex resulting from the inhibition of trypsin by diisopropylphosphofluoridate. The structure is consistent with a mechanism of inactivation that follows an associative pathway, proceeding via a transition state/intermediate in which phosphorus is penta-co-ordinated, forming a trigonal bipyramidal geometry with the phosphonyl donor (p-nitrophenol) and acceptor (Ser70 O gamma atom) in apical positions. A model of this transition state can be accommodated in the active site of beta-lactamase without any steric hindrance. A model of the tetrahedral transition state associated with the acylation step by benzyl penicillin has been derived. Because of the conformational rigidity of the fused rings of penicillin molecules, the orientation of the substrate is fixed once the tetrahedral carbonyl carbon and its ligands are superimposed on the phosphonate group. The outcome is that the carboxylate substituent on the thiazolidine ring forms a salt bridge with Lys234, and the preferred puckering of the ring is that observed in the crystal structure of ampicillin, the so-called "open" conformer. PMID- 8230197 TI - Crystal structure of phospholipase C from Bacillus cereus complexed with a substrate analog. AB - We report the first crystal structure of a complex between PLC from Bacillus cereus (PLCBc) and a competitive inhibitor that is an analog of the natural phospholipid substrate. The structure has been determined at 1.9 A resolution and refined to a final R-factor of 15.7%. The inhibitor binds with its phosphonyl group to the three Zn ions in the active site of the enzyme and is also involved in a hydrogen bonded network including several water molecules and amino acid side-chains which appear to help orient the substrate for productive binding. The interactions within this complex provide some important information regarding the mechanism of PLC-catalyzed hydrolysis of membrane phospholipids. A water molecule, located approximately apical to the diacylglycerol leaving group, seems to be the most likely candidate for the attacking nucleophile which initiates the reaction. PMID- 8230198 TI - Nuclear magnetic resonance structural studies of A.AT base triple alignments in intramolecular purine.purine.pyrimidine DNA triplexes in solution. AB - This study reports on an NMR characterization of an intramolecular purine.purine.pyrimidine (R.RY) triplex containing a central A.AT triple in addition to G.GC and T.AT triples. Our studies establish an A(anti).AT alignment with reversed Hoogsteen pairing stabilized by two N6H ... N hydrogen bonds between adenine residues in the third strand and the purine strand of the duplex. This result, combined with our earlier demonstration of G(anti).GC and T(anti).AT pairing alignments, has provided a definitive experimental approach for differentiating between the base triple pairing possibilities proposed previously by Beal and Dervan for R.RY triplexes. PMID- 8230199 TI - Crystal and molecular structure of d(CGTAGATCTACG) at 2.25 A resolution. AB - Single crystal X-ray diffraction methods have been used to determine the structure of the synthetic DNA dodecamer d(CGTAGATCTACG). The dodecamer presents a new crystal form of B-DNA in the monoclinic system, space group C2 with a = 64.83 A, b = 35.36 A, c = 25.35 A and beta = 92.24 degrees. Structure solution was by molecular replacement and the refinement used a combination of rigid body treatment, molecular dynamics simulated annealing and restrained least-squares methods. The refinement has been concluded with an R-factor of 13.8% for 2120 reflections (78% of what is theoretically available) with F > or = 2 sigma (F) in the resolution range 7.0 to 2.25 A. The asymmetric unit comprises a B-form duplex (24 nucleotides, molecular weight 7.2 kDa), 136.5 water molecules and a single magnesium ion at 50% occupancy. The DNA model has root-mean-square derivations from standard bond lengths of 0.011 A. The central part of the structure is d(GATC), a tetrad sequence involved in methylation and mismatch repair. A comparison with other structures containing such a tetrad indicates polymorphism at the tetrad level. In our structure a narrow minor groove with a pronounced hydration pattern is observed. Such features may be of importance in the recognition of the sequence by specific enzymes. A noteworthy feature of the hydration is the identification of several pentagonal arrangements of hydrogen bonding groups. Details of conformational parameters, base stacking patterns, intermolecular interactions and hydration are presented and comparisons with related structures are given. PMID- 8230200 TI - Mapping the stability determinants of bacterial tyrosyl transfer RNA synthetases by an experimental evolutionary approach. AB - The tyrosyl-tRNA synthetases from Bacillus stearothermophilus (Bst-TyrTS) and Escherichia coli (Eco-TyrTS) are 56% identical in amino acid sequence. To map and characterize the set of interactions that makes Bst-TyrTS more stable than Eco TyrTS, a family of nine hybrid proteins was constructed between the two enzymes. The N-terminal part of each hybrid came from Eco-TyrTS and the C-terminal part from Bst-TyrTS. The stability and activity of these hybrids were estimated by experiments of thermal inactivation and tRNA charging. For all the hybrids, the temperature of half-inactivation in 30 minutes was above 44 degrees C and the rate of charging was at least 40% that of Bst-TyrTS. In general, the temperature of half-inactivation increased and the rate of charging decreased monotonically when the number of residues coming from the more stable and less active Bst-TyrTS increased. As a result, the rate of charging decreased when the temperature of half-inactivation increased. These results show that the sequences and structures of the two enzymes can replace each other locally and still give a stable and active TyrTS, and that the greater stability of Bst-TyrTS is due to cumulative changes of residues scattered along the sequence. They suggest that Bst-TyrTS is more rigid than Eco-TyrTS at low temperature. The existence of a few exceptional hybrids, having stabilities or activities lower than those of the neighbouring hybrids, shows that compensatory changes of residues have occurred between the two sequences during evolution. These exceptions could be explained by the systematic identification of the couples of residues that are in contact in the Bst-TyrTS structure and become heterologous in some hybrids. PMID- 8230201 TI - Refined crystal structure of the seryl-tRNA synthetase from Thermus thermophilus at 2.5 A resolution. AB - The three-dimensional structure of the seryl-tRNA synthetase from Thermus thermophilus has been determined and refined at 2.5 A resolution. The final model consists of a dimer of 421 residues each and 190 water molecules. The R-factor is 18.4% for all the data between 10 and 2.5 A resolution. The structure is very similar to that of the homologous enzyme from Escherichia coli, with an r.m.s. difference of 1.5 A for the 357 alpha-carbon atoms considered equivalent. The comparison of the two structures indicates increased hydrophobicity, reduced conformational entropy and reduced torsional strain as possible mechanisms by which thermostability is obtained in the enzyme from the thermophile. PMID- 8230202 TI - Low energy of activation for amide hydrogen exchange reactions in proteins supports a local unfolding model. AB - Hydrogen exchange reactions of amides in hen egg white lysozyme that are pH dependent and have a low energy of activation have been shown to be in accordance with a reaction mechanism in two steps, an equilibrium step and an exchange step. These results are not in agreement with the model, proposed by C.K. Woodward & B.D. Hilton, known as the penetration model. Therefore our results suggest that this model should be revised. The amide hydrogen/deuterium exchange rates in hen egg white lysozyme were measured at 4 degrees C, 10 degrees C, 15 degrees C and 25 degrees C at pH 7.0 by 1H nuclear magnetic resonance spectroscopy. Activation energies of the exchange reactions in the range from 20 kJ mol-1 to 333 kJ mol-1 were obtained for 32 of the 129 residues in the protein. The amides of lysozyme studied here could be divided into two groups, one group of amides are characterized by an observed amide exchange rate (ko) in the range 10(-4) to 10( 6) s-1, an equilibrium constant k1/k2 close to 10(-5), a low energy of activation (20 to 50 kJ mol-1) and a distance less than 6 A from solvent. The other group of amides are characterized by a ko less than 10(-6) s-1, a k1/k2 close to 10(-7), higher energies of activation (40 to 330 kJ mol-1) and a distance more than 4 A from solvent. In terms of structure the amides of the last group are from the core of the protein. They are typically involved in a hydrogen bond and form part of the secondary structure either as interior alpha-helices or central strands of beta-sheets. The first group consists of amides that are in the shell of the protein between the core and the surface. These amides are typically hydrogen bonded and involved in secondary structure such as external alpha-helices or outer strands of beta-sheets and turns. PMID- 8230203 TI - Crystallization and preliminary X-ray diffraction analysis of the complex of human placental 17 beta-hydroxysteroid dehydrogenase with NADP+. AB - Single crystals of human placental 17 beta-hydroxysteroid dehydrogenase, an enzyme that plays an important role in the interconversion of estrogens, were obtained as the NADP+ complex. These are the first crystals suitable for complete X-ray structure analysis ever reported for a steroid-converting enzyme from a human source. The crystals were grown by vapor diffusion at pH 7.5 with polyethyleneglycol (4000) as the precipitating agent. They have a monoclinic space group C2 and unit cell parameters are a = 123.03 A, b = 45.03 A, c = 61.29 A, and beta = 99.1 degrees. A complete set of diffraction data to 2.9 A has been collected on native crystals. PMID- 8230204 TI - Crystallization of bovine adrenodoxin reductase in a new unit cell and its crystallographic characterization. AB - Plate-like crystals of adrenodoxin reductase from bovine adrenocortical mitochondria were obtained in a new modification. The crystals belong to the monoclinic space group C2 with cell parameters a = 85.94 A, b = 62.64 A, c = 128.55 A and beta = 99.8 degrees. There are two molecules in the asymmetric unit. The crystals diffract to better than 3.0 A resolution at 4 degrees C. PMID- 8230205 TI - In vitro binding of the pleiotropic transcriptional regulatory protein, FruR, to the fru, pps, ace, pts and icd operons of Escherichia coli and Salmonella typhimurium. AB - Evidence has been presented suggesting that the fructose repressor, FruR, is a pleiotropic transcriptional regulatory protein controlling the expression of numerous operons concerned with carbon metabolism in Escherichia coli and Salmonella typhimurium. We have conducted in vitro DNA binding studies to ascertain the nature of the DNA sequences to which FruR binds. Employing both DNA band migration retardation and DNase I footprint analyses, FruR was found to bind to two operators within the regulatory region preceding the structural genes of the fructose operon, fruB(MH)KA. These two operators, O1 and O2, comprise nearly identical palindromes of 12 bp with a half-site of TGAAAC. The binding of FruR to these inverted repeats was found to be reversed by inclusion of micromolar concentrations of fructose-1-phosphate. The two operators are located between the single putative promoter of the fructose operon and the translational initiation site of the fruB gene. Other regulated operons were shown to bind FruR to a single site upstream of the first structural gene as follows: (1) ppsA (positive regulation); (2) icd (positive regulation); (3) aceB (positive regulation); and (4) pts (negative regulation). In all cases, low concentrations of fructose-1 phosphate displaced the protein from the DNA. The binding sites were determined, and a FruR consensus sequence was established. Computer searches revealed the presence of this sequence in numerous functionally diverse operons, implying that FruR is a global transcriptional regulatory protein in enteric bacteria. PMID- 8230206 TI - Characterization of a set of integration host factor mutants deficient for DNA binding. AB - Integration host factor, IHF, is a sequence-specific DNA-binding and DNA-bending protein composed of two related but non-identical subunits. We report the isolation and characterization of hydroxylamine-induced loss-of-function mutations in the genes encoding the IHF subunits. To screen for mutants that preserve proper folding of IHF, clarified extracts were prepared from each mutant and were assayed for production of each subunit by immunoblotting and for formation of heterodimers by chemical cross-linking and subsequent immunoblotting. Extracts from mutants that met these criteria were found to bind a specific IHF site weakly if at all. These alleles therefore identify candidates for residues that may affect the DNA-binding surfaces of IHF. When projected onto the known tertiary structure of the closely related HU protein, these residues are found at the surface; however, with the exception of a single residue, different regions of the protein are implicated in each subunit. This suggests that, despite their homology, each subunit of IHF directs DNA recognition and binding in a distinct manner. To confirm the significance of the differential location of these mutations, we introduced in each subunit alterations that had been isolated as loss-of-function mutations at the corresponding position in the other subunit. In general, the engineered mutants have phenotypes that are strikingly different from those of their hydroxylamine-induced counterparts. In particular, most of the site-directed mutant IHF proteins form or maintain IHF:DNA complexes more readily than mutants that have the same change in the other subunit and were isolated as loss-of-function mutants. We discuss the positions of the mutant amino acid residues as they relate to a proposed molecular model of an IHF:DNA complex. PMID- 8230207 TI - DNA recognition by the EcoK methyltransferase. The influence of DNA methylation and the cofactor S-adenosyl-L-methionine. AB - The methyltransferase of the EcoK type I restriction/modification system is trimeric, M2S1, where the S subunit determines the sequence specificity of the enzyme. The methyltransferase has a strong preference for hemimethylated substrate DNA and, therefore, we have investigated the effect of the methylation state of DNA on binding by the enzyme, together with the effects on binding of the cofactor S-adenosyl-L-methionine. Our results indicate that the methyltransferase has two non-interacting S-adenosyl-L-methionine binding sites, each with a dissociation constant of 3.60 (+/- 0.42) microM determined by equilibrium dialysis, or 2.21 (+/- 0.29) microM determined by the displacement of a fluorescent probe. Ultraviolet light-induced crosslinking showed that S adenosyl-L-methionine binds strongly only to the modification (M) subunits. Changes in the sedimentation velocity of the methyltransferase imply a protein conformational change due to S-adenosyl-L-methionine binding. Gel retardation results show that the binding of S-adenosyl-L-methionine to the methyltransferase enhances binding to both specific and non-specific DNAs, but the enhancement is greater for the specific DNA. Differences in binding affinities contribute to the recognition of the specific nucleotide sequence AAC(N)6GTGC by the methyltransferase in preference to a non-specific sequence. In contrast, although the complexes of unmodified and hemimethylated DNAs with the methyltransferase have different mobilities in non-denaturing gels, there appears to be no contribution of binding affinity to the distinction between these two substrates. Therefore, the preference for a hemimethylated substrate must be due to a difference in catalysis. PMID- 8230208 TI - Negative co-dominant inhibition of recA protein function. Biochemical properties of the recA1, recA13 and recA56 proteins and the effect of recA56 protein on the activities of the wild-type recA protein function in vitro. AB - We have investigated the biochemical properties of several Escherichia coli mutant recA proteins that display a null phenotype. These are the recA1, recA13 and recA56 proteins, each of which carries a single missense mutation. These proteins all share a common defect which is the inability to adopt the high affinity DNA binding state normally elicited by the nucleotide cofactor ATP. Consequently, other than the ability to bind ssDNA, they possess none of the in vitro enzymatic activities of recA protein. However, each protein has characteristics that are unique, leading to the conclusion that the observed mutant phenotypes arise through fundamentally different mechanisms. Despite the magnitude of these defects, the recA56 protein is able to differentially inhibit various activities of wild-type recA protein. Incorporation of recA56 protein into a presynaptic filament with the wild-type recA protein does not affect the ability of the wild-type protein to hydrolyze ATP, as judged by the turnover number (kcat), provided that the ssDNA concentration is not limiting; however, the affinity of wild-type recA protein for ATP is lowered by the presence of recA56 protein. Similarly, the ability to cleave lexA protein is only modestly inhibited. However, both the ability to compete with SSB protein for ssDNA binding sites and the DNA strand exchange activity of wild-type recA protein are severely inhibited by the presence of recA56 protein. These results suggest that individual monomeric components of the recA protein-DNA filament are translated through protein-protein contacts to become macroscopic properties of the filament. PMID- 8230209 TI - Assembly of multimeric proteins. Effect of mutations in the alpha-subunit on membrane assembly and activity of pyridine nucleotide transhydrogenase. AB - The pyridine nucleotide transhydrogenase of Escherichia coli is an inner membrane protein of two different subunits (alpha and beta). It functions as a proton pump. The highly hydrophilic carboxy-terminal tail of ten amino acid residues in the alpha-subunit determines the correct folding and proper assembly of the beta subunit leading to a functional enzyme. Premature termination of the alpha subunit six amino acid residues from the carboxy-terminal end abolishes the activity completely. Although the two subunits are still assembled into the membrane, the conformation of the beta-subunit is perturbed. Systematic truncation and site-directed substitutions revealed that at least one positive charge in the carboxy-terminal region is required for efficient assembly of the two subunits to give a functional enzyme, while a phenylalanine residue, essential for activity, has no apparent effect on the extent of assembly of the two subunits. PMID- 8230210 TI - A regulatory cascade in the induction of rhaBAD. AB - The RhaS and RhaR regulatory proteins are encoded in the Escherichia coli L rhamnose gene cluster. We used complementation analysis and DNA mobility shift assays to show that RhaR is not the direct activator of the L-rhamnose catabolic operon, rhaBAD. An in-frame deletion of rhaS (rhaS-rhaR+) eliminated expression from the rhaBAD promoter, pBAD, while overexpression of rhaS greatly speeded the normally slow induction of transcription from pBAD. Expression from pBAD in a coupled transcription-translation assay was only detected when rhaS+ DNA was added to allow synthesis of RhaS protein. RhaS thus appears to be the direct L rhamnose-specific activator of rhaBAD expression. Deletion mapping located the binding site for the L-rhamnose-specific regulator to a region overlapping position -70 relative to the rhaBAD transcription start site. Deletion mapping and DNA mobility shift assays located a CRP binding site just upstream from the binding site for the L-rhamnose-specific regulator. Quantitative primer extension analysis showed that induction of both the rhaBAD and rhaSR messages was unusually slow, requiring 40 to 50 minutes to reach a steady-state level. Induction of rhaBAD apparently involves a regulatory cascade in which RhaR first induces rhaSR expression, then RhaS accumulates and induces rhaBAD expression. PMID- 8230211 TI - Temperature-sensitive intermediate filament assembly. Alternative structures of Xenopus laevis vimentin in vitro and in vivo. AB - In assembly assays of intermediate filaments (IFs) from vimentin of the amphibian species Xenopus laevis we have observed the formation of so far unknown structures at temperatures above 28 degrees C. Upon assembly in vitro at temperatures above 34 degrees C massive aggregates, partly with a protofilamentous substructure, were found and their formation correlated with drastically reduced end-viscosity. Large spheroidal, dense aggregates with a complex suborganization were also seen to form at 37 degrees C in the cytoplasm of living mammalian cells devoid of endogenous vimentin upon transfection with cDNA encoding the amphibian vimentin, and this was also true for vimentin forced to accumulate in the nucleoplasm by the introduction of a "nuclear localization signal". Upon shift from the non-permissive (37 degrees C) to the permissive (28 degrees C) temperature, such aggregates of non-IF vimentin structures gradually disappeared and a normal-looking IF meshwork formed. The results, which are discussed in relation to other structures assembled by IF proteins, indicate a marked thermosensitivity in the amino acid sequence of the vimentin which seems to have been reduced during evolution of warm-blooded animals. They further show that members of the multigene gene family of IF proteins can occur in structures totally different from IFs. PMID- 8230212 TI - Rules that govern tRNA identity in protein synthesis. AB - The specificity of tRNA in protein synthesis depends not only on its recognition of the codon in the mRNA, but also on its recognition of the correct aminoacyl tRNA synthetase enzyme. The specificity of tRNA in aminoacylation (tRNA identity) depends on the tRNAs productive interaction with the correct enzyme and non productive interaction with all other enzymes. Although extensive regions of the tRNA interact with the enzyme, only a small number of nucleotides comprise the major determinants of tRNA identity. They often lie in the same positions (acceptor end and anticodon, and variable pocket less often) in different tRNAs. Therefore, a determinant in a given tRNA simultaneously ensures both productive and non-productive interactions with the respective enzymes. Specificity for the acceptor end of the tRNA is achieved, in part, by the specific amino acid sequence within protein binding pocket domains that are part of all aminoacyl tRNA synthetases. These domains also bind the other two substrates of the enzyme, amino acid and ATP. Specificity for the anticodon and variable pocket of the tRNA is more idiosyncratic. Irrespective of their location in the tRNA, the determinants either interact directly with the enzyme or give the tRNA a conformation for a complementary fit with the enzyme. PMID- 8230213 TI - Chimeric human cholinesterase. Identification of interaction sites responsible for recognition of acetyl- or butyrylcholinesterase-specific ligands. AB - Acetyl- and butyrylcholinesterases (AChE, BuChE) from various species differ in their substrate specificities and sensitivities to a wide range of inhibitors, yet display conserved sequence, structure and catalytic properties. To determine features that confer these selective properties, residues 58 through 133 of recombinant human BuChE were replaced with the corresponding sequence from human AChE. The replaced region (> 60% identity) spans the Asp70 residue, important for ligand interactions, and the choline binding site, and introduces differences of charge and hydrophobicity in the outer rim and on the surface of the active site gorge. Expressed in microinjected Xenopus laevis oocytes, the resultant chimera retained the catalytic activity, substrate specificity and the Km value toward butyrylthiocholine characteristic of BuChE. Further, it did not acquire substrate inhibition, which is unique to AChE, although it lost the property of substrate activation, characteristic of BuChE. Moreover, the chimera resembled BuChE in its sensitivity to succinylcholine and physostigmine, but acquired the AChE-like sensitivity to echothiophate and iso-OMPA, and displayed an intermediate pattern of inhibition, more similar to that of AChE than of BuChE, toward bambuterol, dibucaine and BW284C51. These findings demonstrate that the exchanged residues are involved in inhibitor recognition, but not in substrate distinction and in direct catalysis. Furthermore, substrate interaction with the exchanged domain may mediate structural changes leading to substrate activation in BuChE and inhibition in AChE. The two AChE-specific aromatic tyrosine residues positioned near Asp70 within this region are hence implicated in the peripheral anionic site of cholinesterases, which is involved in the recognition of various ligands. PMID- 8230214 TI - Effect of positive supercoiling on DNA compaction by nucleosome cores. AB - We have used the analytical ultracentrifuge to measure the ability of histone octamers to compact DNA as a function of DNA supercoiling. Plasmid DNA (3.25 kb) was prepared at various linking numbers (delta Lk), ranging from -35 to +8. Relaxed plasmid (delta Lk = 0) was the least compact. We reconstituted a fixed number of nucleosome cores (either 11 or 13) on these DNAs. The dependence of the frictional coefficient of delta Lk showed that the reconstitute with an initial number of negative supercoils equal to the number of nucleosome cores was the least compact, as expected if each nucleosome core formed requires the constraint of one negative supercoil, resulting in relaxed linker DNA. With DNAs containing an initial number of negative supercoils unequal to the number of nucleosome cores, reconstitutes contained either negative or positive unconstrained supercoils. Reconstitutes with the same number of unconstrained supercoils, whether positive or negative, have similar frictional coefficients and are, therefore, compacted to similar degrees. We conclude that nucleosome cores compact positively and negatively supercoiled DNA equally well. Thus, nucleosome cores formed on positively supercoiled DNA with a superhelical density as high as +0.07 are not significantly unfolded. PMID- 8230215 TI - Molecular recognition mediated by bound water. A mechanism for star activity of the restriction endonuclease EcoRI. AB - Many restriction endonucleases such as EcoRI lose some specificity for their recognition sequence under certain buffer conditions. The cause of this disruption of accurate protein-DNA recognition has never been explained. By cleaving DNA with EcoRI in the presence of several osmolytes, we show that the extent of this EcoRI "star activity" depends strongly upon osmotic pressure. The loss of specificity accompanying decreased water activity implies a role for one or more water molecules in recognition of specific sequences of DNA. Water mediation may constitute a general motif for sequence-specific DNA recognition by restriction enzymes and other DNA-binding proteins. PMID- 8230216 TI - The kin28 protein kinase is associated with a cyclin in Saccharomyces cerevisiae. AB - Kin28 is an essential serine-threonine kinase of Saccharomyces cerevisiae. Multicopies of a novel cyclin gene, CCL1, are able to suppress the thermosensitivity of two kin28-ts mutants. The CCL1 gene is not cyclically transcribed, yet its product is also essential for cell proliferation. Furthermore, when overexpressed under high expression promoter, Cell is able to replace G1 function of Cln cyclins. Cell and Kin28 are physically associated in vivo. Therefore, like p34CDC28/cdc2, Kin28 may be a cyclin dependent kinase which is required for cell proliferation. PMID- 8230217 TI - GEBF-I activates the Drosophila Sgs3 gene enhancer by altering a positioned nucleosomal core particle. AB - The enhancer region of the Drosophila melanogaster ecdysone-regulated glue gene, Sgs3, shows dramatic modifications of chromatin structure in strict correlation with changes in gene expression during development. We show that there is a positioned nucleosomal core particle over the enhancer which is displaced or disrupted during gene activation. This transition is prevented in Drosophila larvae mutated in the ecdysone-dependent 2B5 locus, in which Sgs3 is inactive and GEBF-I, a Glue Enhancer Binding Factor, is missing. We have defined the GEBF-I binding sites in vitro and shown that mutation of these sequences abolishes the enhancer activity in vivo. This combined in vitro and in vivo approach reveals new aspects of the dynamic organization of a regulatory element during development and highlights the potential of this model for studies of the relation between chromatin structure and gene activity. PMID- 8230218 TI - Monitoring of the cooperative unfolding of the sunY group I intron of bacteriophage T4. The active form of the sunY ribozyme is stabilized by multiple interactions with 3' terminal intron components. AB - We have studied the mechanism by which the 3' terminal domain of the sunY intron of bacteriophage T4 activates the group I ribozyme core of this intron, from which it is separated by some 800 nucleotides. As shown by monitoring either UV absorbance or self-splicing reaction kinetics as a function of temperature, intron transcripts undergo highly cooperative unfolding/inactivation upon heating: the two methods yield similar estimates of the thermodynamic parameters associated with this process. Such cooperativity makes it possible in turn to assess the energetic contribution of specific interactions to the overall structure, by comparing the sensitivity to heat inactivation of molecules carrying various nucleotide substitutions. By combining this approach with chemical modification, we have probed several proven or putative interactions between the core and 3' terminal domain of the intron and conclude that the role of the 3' terminal domain is to stabilize the active form of the ribozyme. Interestingly, the P9.0 interaction, which brings 3' terminal nucleotides next to the core site that binds the guanosine cofactor of the self-splicing reaction, is now shown to be composed in fact of two distinct pairings. An isolated base-pair (P9.0a), involving a residue located only six nucleotides upstream of the 3' splice site, participates in the stabilization of the ribozyme and appears to persist during the second stage of self-splicing (exon ligation). In contrast, formation of the previously demonstrated P9.0b pairing, which involves the two penultimate intron nucleotides, contributes no additional stability and results in no detectable rearrangement of the core structure. Implications for the concept of a static ribozyme are discussed in the light of a slightly revised three-dimensional model of the sunY intron. PMID- 8230219 TI - Lipid-protein interactions in crystals of plant light-harvesting complex. AB - Two different thylakoid lipids are specifically associated with the light harvesting complex of photosystem II (LHC-II). Digalactosyl diacyl glycerol (DGDG) binds to the isolated complex but can be removed by mild detergent treatment and anion-exchange chromatography. Removal of this lipid renders the complex unable to form two-dimensional or three-dimensional crystals. The ability to crystallize is completely restored by addition of pure DGDG, at a ratio of about four molecules per polypeptide for three-dimensional crystals, suggesting several binding sites at the periphery of the trimeric complex. Two-dimensional crystals of purified protein grown in the presence of DGDG are more highly ordered than those obtained from the unfractionated complex. The other lipid, phosphatidyl glycerol (PG), binds more firmly and cannot be removed with non ionic detergent. Complete delipidation of LHC-II can be achieved either with phospholipase or by proteolytic cleavage of 49 amino acid residues at the N terminus. Both treatments dissociate the native, trimeric complex into monomers. This indicates that PG is directly involved in the formation of trimers, which are a prerequisite for two-dimensional and three-dimensional crystallization. Both lipids are therefore present in two-dimensional and three-dimensional crystals and have distinct roles in the structure of the complex. PMID- 8230220 TI - Domain closure in lactoferrin. Two hinges produce a see-saw motion between alternative close-packed interfaces. AB - Lactoferrin is an iron transport protein. Upon binding iron, the two domains in the N-terminal half of the molecule move together. Previous work has shown that this domain closure involves two hinges. Using the newly refined structure of the open form, the structural mechanism underlying this motion is analyzed here in detail. Upon closure the domains rotate 54 degrees essentially as rigid bodies. The axis of rotation passes through the two beta-strands linking the domains. These strands contain hinges in the sense that three large torsion angle changes are responsible for the bulk of the motion while smaller torsion angle changes in neighboring residues are responsible for the remainder of the motion. The rotation axes of these three torsion angle changes are nearly parallel to the axis of the overall 54 degrees rotation, so the local motion in the hinges can be directly related to the overall motion. A crucial feature of the hinge residues is that they have very few packing constraints on their main-chain atoms. The domains make different packing contacts with each other in the open and closed forms. These contacts form two interdomain interfaces arranged on either side of the hinges. Pivoting about the hinges produces a see-saw motion between the two interfaces. That is, when the domains close down, residues in the interface on one side of the hinges become buried and close-packed and residues on the other side become exposed. The situation is reversed when the domains open up. Lactoferrin provides a particularly clear example of the general features of hinged domain motion. It is compared to other instances of hinged domain closure and contrasted with instances of shear domain closure, where the overall motion is a summation of many small sliding motions between close-packed segments of polypeptide. PMID- 8230221 TI - Molecular dynamics with weighted time-averaged restraints for a DNA octamer. Dynamic interpretation of nuclear magnetic resonance data. AB - For conformationally flexible molecules in solution, NMR-derived distance restraints are time-averaged. In contrast to deriving structures from NMR distance constraints via conventional restrained molecular dynamics (rMD), the range of conformational flexibility may be better represented by MD simulations using weighted time-averaged restrains (MD-tar). This approach has been utilized for dynamic structure refinement of the DNA octamer [d(GTATAATG].[d(CATATTAC)] which contains the Pribnow box consensus sequence. An improved set of distance restraints was calculated via complete relaxation matrix analysis utilizing the solution structure of the octamer which was determined previously by rMD as a starting model. MD-tar calculations were performed with the program AMBER4 under various conditions, some including explicit solvent. All trajectories generated via MD-tar exhibited reduced constraint energies and average deviations for the distances compared to standard rMD. Quantitative comparison with experimental data, i.e. two-dimensional NOE intensities and COSY-derived coupling constants yielded a significant improvement for MD-tar simulated structures relative to rMD derived structures. The conformational envelope of the MD-tar structures is wider than that from rMD and even unrestrained MD. Explicit solvent force-fields tightened the conformational envelope, leading to even better agreement with experimental data. All MD-tar simulations exhibit sugar repuckering for basically all residues, yielding a minor population in the low N-region and one or more S conformers. For most backbone torsion angles, one or more minor conformers were found, while the major conformations generally coincided with those obtained in standard rMD. Distributions of helical parameters for MD-tar trajectories are rather symmetric but slightly broader than those for rMD. Average values and associated standard deviations are discussed with respect to sequence-dependent variations. All trajectories obtained with an explicit solvent force field exhibited a narrower minor groove compared to in vacuo calculations. PMID- 8230222 TI - Two crystal structures of a potently sweet protein. Natural monellin at 2.75 A resolution and single-chain monellin at 1.7 A resolution. AB - Two refined structures of the sweet-tasting protein monellin are presented. The structure of natural monellin has been refined at 2.75 A resolution. The final model consists of four monellin molecules in the asymmetric unit, encompassing 3136 non-hydrogen atoms. The crystallographic R-factor is 0.193 for the 8853 reflections between 6.0 A and 2.75 A resolution, and the root-mean-square deviations from ideality are 0.017 A for bond lengths and 3.6 degrees for bond angles. The refined structure generally confirms, with some difference in detail, the initial backbone model of monellin that was based on 3.0 A resolution data. Single-chain monellin (scm) was genetically engineered by fusing the two chains of monellin into a single 94-residue polypeptide. Using the refined monellin coordinates as a search model, the crystal structure of scm has been solved with the techniques of molecular replacement, and has been refined against data to 1.7 A resolution. The final model consists of two scm molecules per asymmetric unit, and includes 137 bound water molecules. The crystallographic R-factor for this model is 0.174 for the 15,053 reflections (magnitude of FO magnitude of > 2 sigma (FO)) between 6.0 A and 1.7 A resolution. The root-mean-square deviations from ideal bond lengths and angles are 0.015 A and 2.86 degrees, respectively, and the average coordinate error is approximately 0.2 A, as estimated from a Luzzati plot. The error in the model was also estimated by comparing the two molecules in the asymmetric unit. The most significant differences between the two molecules occur in loop regions and at the C terminus of the protein, and are generally correlated to differences in crystal packing contacts. Linking the two chains of monellin has not substantially altered the structure beyond the region immediately surrounding the new peptide bond. Like natural monellin, the conformation of scm is dominated by a 17-residue alpha-helix folded into the concave side of a twisted, five-strand anti-parallel beta-sheet. We expect that the availability of a high-resolution structure of scm, along with the convenience of producing site-specific mutants of this protein, will make scm a good model with which to probe the structural basis of sweetness. PMID- 8230223 TI - Three-dimensional structure in solution of the calcium channel blocker omega conotoxin. AB - The 27 amino acid residue polypeptide omega-conotoxin GVIA, from venom of the cone shell Conus geographus, blocks neuronal voltage-activated calcium channels at picomolar concentrations. The three-dimensional structure in aqueous solution of synthetic omega-conotoxin has been determined from two-dimensional 1H n.m.r. data recorded at 600 MHz. Structural constraints consisting of interproton distances inferred from NOEs and dihedral angles from spin-spin coupling constants were used as input for distance geometry calculations with the program DSPACE. The structures were then refined using back-calculation of NOESY spectra. The family of structures obtained in this way is well defined by the n.m.r. data, the best 12 structures having pairwise root-mean-square differences of 0.68 (+/- 0.15) A over the backbone heavy atoms (N, C alpha and C) and 1.15 (+/- 0.17) A over all heavy-atoms. The molecule adopts a compact structure consisting of a small, triple-stranded, anti-parallel beta-sheet and several reverse turns. All three tyrosine residues are located on the molecular surface, which is noteworthy for its abundance of side-chain hydroxyl groups. There is no negatively charged group in conotoxin, but the five positively charged groups are distributed in three small patches on the surface, one of which, made up of the ammonium moieties of the N terminus and Lys2, may contribute to the receptor-binding surface of the molecule. An isomer of conotoxin with the same amino acid sequence, but different disulfide pairings, has also been investigated. Its structure is less well ordered than that of native conotoxin and it shows significant heterogeneity, probably as a result of cis-trans isomerism preceding hydroxyproline residues. PMID- 8230224 TI - Atomic structure of a cytochrome c' with an unusual ligand-controlled dimer dissociation at 1.8 A resolution. AB - The crystallographic structure of cytochrome c' from the purple phototrophic bacterium Chromatium vinosum (CVCP) has been determined at 1.8 A resolution using multiple isomorphous replacement. The molecule is a dimer, with each 131-residue chain folding as a four-helical bundle incorporating a covalently bound heme group at the core. This structure is the third of the ubiquitous cytochromes c' to be solved and is similar to the known structures of cytochrome c' from R. molischianum (RMCP) and R. rubrum (RRCP). CVCP is unique in exhibiting ligand controlled dimer dissociation while RMCP and RRCP do not. The Tyr16 side-chain, which replaced Met16 in RMCP and Leu14 in RRCP, is parallel to the heme plane and located directly above the sixth ligand site of the heme Fe. Any ligand binding to this site, such as CO or CN-, must move the Tyr16 side-chain, which would be expected to cause other conformational changes of helix A, which contributes to the dimer interface, and consequently disrupting the dimer. Thus, the crystallographic structure of CVCP suggests a mechanism for dimer dissociation upon ligand binding. The dimer interface specificity is due to a lock and key shape complementarity of hydrophobic residues and not to any charge complementarity or cross-interface hydrogen bonds as is common in other protein protein interfaces. The co-ordinates have been deposited in the Brookhaven Data Bank (entry P1BBH). PMID- 8230225 TI - Structure of the complex of lac repressor headpiece and an 11 base-pair half operator determined by nuclear magnetic resonance spectroscopy and restrained molecular dynamics. AB - The structure of the complex of lac repressor headpiece and an 11 base-pair lac half-operator has been determined by NMR spectroscopy and restrained Molecular Dynamics calculations. In total 508 distances were derived from two-dimensional nuclear Overhauser enhancement measurements, 260 of which are within the headpiece, 212 within the operator and 36 between operator and headpiece. An equilibrium restrained Molecular Dynamics calculation of the complex in aqueous solution, spanning 85 picoseconds, has been used to analyze the structure. Configuration sampling by an annealing procedure has been undertaken as well in order to estimate the precision of the structure determination. Our data confirm the results of previous two-dimensional NMR studies that the orientation of the recognition helix of lac repressor in the major groove of DNA with respect to the operator dyad axis is opposite to the orientation found in complexes of other DNA binding proteins of the helix-turn-helix class. We find a number of tight contacts between the protein and the operator that are in agreement with the available genetic and biochemical data. The anchoring of lac headpiece on the operator is similar to that of other repressors. Other features are unique for lac headpiece: relative few direct hydrogen bonds between side-chains and bases; extensive apolar contacts; many direct and water-bridged contacts to phosphates from residues in or close to the recognition helix. Overall, an interconnected set of interactions is observed, involving base-specific contacts, phosphate contacts, intra-protein and water-bridged hydrogen bonds. Several of these interactions appear to be dynamic, i.e. fluctuating in time, rather than static. PMID- 8230226 TI - Anion binding sites in protein structures. AB - An analysis of the binding of 52 sulfate and phosphate ions in 34 protein structures has been carried out. On average an oxyanion is held by 7(+/- 3) hydrogen bonds, of which the contribution of the protein is 5(+/- 3) (the rest coming from water). Arg and the peptide NH group are the most common ligands, other basic and polar side-chains, including Asp and Glu, have their own shares. The negative charge on the ion is not neutralized by the protein in all the cases. The geometry of the ligand (L) with reference to a SO4 or PO4 is such that the angle S/P-O..L is 128(+/- 15)degrees and 96(+/- 15)degrees for the two classes of interactions distinguished by the existence of only one or more than one hydrogen bond connecting the ligand atom and the anion. The binding belonging to the former category is usually in a staggered orientation with respect to the anion-tetrahedron; those in the latter class are close to the eclipsed conformation. An anion is held in a very limited space close to the NH direction of a peptide group; the strongly localized nature of the interaction imparts rigidity to the binding. Both local steric and electrostatic elements favor the peptide group to be at the N terminus of a helix. Additional ligands can be supplied by the polypeptide chain preceding the helical N terminus. beta-Turns are also propitious for the location of ligands. There are patterns in the binary and ternary interactions involving various anions. One of the constituents of the ligand peptide group is likely to be a Gly; with no side-chain this residue offers the minimum steric hindrance to the bound anion. In 50% of the cases with a Ser, a Thr or a basic residue occupying one of these positions, the side-chain also acts as a hydrogen bond donor. Gly is also likely to occur next to a side chain ligand located in a beta-turn. About a third of the anions have ligands from a crystallographic or molecular symmetry-related position. PMID- 8230227 TI - Dissecting the structure of a partially folded protein. Circular dichroism and nuclear magnetic resonance studies of peptides from ubiquitin. AB - The nature and interaction of structural elements in a partially ordered form of ubiquitin, the A-state, which is populated at low pH in 40 to 60% aqueous methanol, have been investigated. Two synthetic peptides have been studied under the same conditions: U(1-21), corresponding to the N-terminal beta-hairpin in the native (N) and A-states of ubiquitin and U(1-35), which includes this hairpin plus an alpha-helix. Circular dichroism studies indicate that, although these peptides are largely unfolded in water, their structural content in 30 and 60% methanol is comparable with the corresponding native secondary structure. Sequence-specific assignments of the 1H n.m.r. spectra of U(1-35) in aqueous methanol and subsequent secondary structure determination confirm the conservation in detail of native-like secondary structure. Corresponding resonances in spectra of U(1-35), U(1-21) and the A-state itself were found to have closely similar chemical shifts, suggesting that the beta-hairpin exists independently in the partially folded protein, with little or no influence from the rest of the molecule. This is confirmed by the virtual absence in nuclear Overhauser enhancement spectroscopy and rotating frame nuclear Overhauser enhancement spectroscopy spectra of nuclear Overhauser enhancement effects between structural elements. c.d. and n.m.r. evidence suggests that structure in the C-terminal half of the molecule in the A-state is largely non-native. Thus, although methanol is necessary to assure its stability in the absence of wider native interactions, the structure of the beta-hairpin, including the register of its hydrogen bonding, appears to be determined entirely by its own sequence. This intrinsic structural preference in the first part of the ubiquitin sequence is much stronger than in the C-terminal half, a conclusion reflected in the results from a variety of secondary structure prediction algorithms. PMID- 8230228 TI - Crystallization and preliminary crystallographic characterization of bacteriophage T4 baseplate protein encoded by gene 9. AB - The structural protein, gene product 9 (gp9), of bacteriophage T4 controls baseplate expansion at the first steps of virus attachment onto its host bacterial cell with subsequent tail contraction. Gp9, which has an M(r) of 30.8 kDa and contains 287 amino acids, has been purified from a recombinant Escherichia coli strain and crystallized at 25 degrees C using the hanging drop vapor diffusion method at pH 4.0 with ammonium sulfate as precipitant. The crystals of gp9 belong to the space group R32 with hexagonal cell dimensions a = b = 86.5 A and c = 156.2 A and diffract X-rays to at least 2.7 A. There is one molecule per asymmetric unit. PMID- 8230229 TI - Cocrystals of Escherichia coli trp repressor bound to an alternative operator DNA sequence. AB - Cocrystals of a 2:1 complex of trp repressor dimers with a DNA duplex containing a single, central operator half-site sequence are described. Crystals with different morphologies grew under diverse crystallization conditions within days to weeks by hanging-drop vapor-diffusion. Twinned rods split along their longitudinal cleft produce single crystals with space group C2 and unit cell dimensions a = 112.34 A, b = 90.16 A, c = 58.65 A and beta = 113.92 degrees. The crystals diffract to 2.4 A and are thus suitable for structural analysis by X-ray diffraction. Several heavy-atom derivative cocrystals have been obtained with iodouridine-substituted DNAs. PMID- 8230230 TI - Preliminary X-ray crystallographic studies of ribonuclease I from Escherichia coli. AB - Single crystals of ribonuclease I from Escherichia coli have been obtained by the vapor diffusion method. The crystals belong to the tetragonal space group P4(1)2(1)2 or its enantiomer P4(3)2(1)2 and have cell parameters a = b = 119.01 A and c = 34.40 A. There is one 27,000 dalton monomer in the asymmetric unit. The crystals diffract to beyond 3.0 A resolution. PMID- 8230231 TI - Crystallization of Old Yellow Enzyme illustrates an effective strategy for increasing protein crystal size. AB - Old Yellow Enzyme (OYE) was the first flavoenzyme discovered, and has been widely used as a model flavoenzyme. Three crystal forms have been grown from natural protease-nicked OYE and one has been grown from intact recombinant OYE. The recombinant OYE crystals are best suited for structure determination and grow from a solution containing polyethylene glycol and MgCl2. They belong to space group P4(3)2(1)2 with unit cell dimensions a = b = 142.88 A, c = 43.01 A and have a single OYE chain per asymmetric unit. In order to optimize the production of large single crystals, we have carried out a systematic study of crystal size versus drop volume. Crystallographic lore holds that the volume of the medium in which the crystals are grown should have a significant effect on the final size of the crystals produced. However, a systematic investigation of the degree to which the volume of the crystallization sample affects the final size of the crystals has not been reported. In order to investigate this phenomenon, drops were set up in both hanging and sandwich geometry at a variety of drop volumes, but otherwise identical crystallization conditions. A 60-fold increase in drop volume produced a 730-fold increase in crystal volume, and a simultaneous increase in the effective diffraction limit of the crystals from near 2.5 A to well beyond 2.0 A resolution. The dramatic crystal volume increase appears to be due both to the increased amount of protein and the slower equilibration rates associated with larger drops. Our results also emphasize that the quality of diffraction data may often be limited by crystal size rather than intrinsic crystal order even in the case of reasonably sized (200 to 400 microns) crystals. PMID- 8230232 TI - Nucleotide sequence analysis of four genes, hupC, hupD, hupF and hupG, downstream of the hydrogenase structural genes in Bradyrhizobium japonicum. AB - The nucleotide sequence of a 2.2 kb region downstream of the hydrogenase structural genes in Bradyrhizobium japonicum was determined. Four genes encoding predicted polypeptides of 27.8 (HupC), 21.4 (HupD), 10.6 (HupF) and 15.8 (HupG) kDa were identified, of which the first three probably belong to the same operon as the hup structural genes, hupS and hupL. HupC is homologous to the hydrophobic polypeptides with four potential transmembrane regions that are encoded by open reading frames following the hydrogenase structural genes in Rhodobacter capsulatus, Escherichia coli, Azotobacter vinelandii, Wolinella succinogenes, Rhizobium leguminosarum and Alcaligenes eutrophus. Also HupD, HupF and HupG are homologous to genes involved in processing, maturation, functioning and regulation of hydrogenase activity in various hydrogen-oxidizing bacteria. PMID- 8230233 TI - Microtopography of Nippostrongylus brasiliensis (Nematoda: Heligmosomatidae): free-living larval stages. AB - Microtopographic features of the various growth stages of the three free-living larval stages of the rat hookworm Nippostrongylus brasiliensis (Nematoda) were surveyed by scanning electron microscopy. These worms have a rounded anterior end and an elongated tail. Cuticular annulations were observed along the body, which also bore two ribbon-like lateral alae. Two rings of six lip-like lappets were observed around the triradiate oral opening in all larval stages. The cephalic space contained two lateral amphidial pits. The excretory pore in the third anterior part was observed in a ventral view of the larvae. No deirids were observed. The anus with a crescent-shape opening was located posteriorly. Phasmidial apertures, only observed in the third-stage larvae, opened on the lateral alae in the tail region. PMID- 8230234 TI - Microtopography of Nippostrongylus brasiliensis (Nematoda: Heligmosomatidae): parasitic larval stages and adults. AB - Specimens of the rat hookworm, Nippostrongylus brasiliensis (Nematoda) were recovered from lungs (third- and fourth-stage larvae) and intestine (fourth-stage larvae and adults). The following features were studied in the different stages by scanning electron microscopy: cephalic structures, especially sense organs, synlophe, cervical region, and caudal part. The main differences between the third and fourth stages concerned the lip-like structures around the oral aperture, the appearance of the cephalic space with the presence of a cephalic cap in fourth-stage larvae, the pattern of longitudinal ridges, and sexual differentiation. Pore-like papillae, not seen in third-stage larvae, developed in later stages. Deirids were observed only in adults, and phasmids were poorly discerned. Some of these morphological features, such as the cephalic sense organ apertures and cuticle pores and micropores, can be observed only by scanning electron microscopy. The possible functions of these different structures and their relationship with the behavior of the worms during their life cycle are discussed. PMID- 8230235 TI - Histochemical and myosin composition of vampire bat (Desmodus rotundus) pectoralis muscle targets a unique locomotory niche. AB - The vampire bat pectoralis muscle contains at least four fiber types distributed in a nonhomogeneous pattern. One of these fiber types, here termed IIe, can be elucidated only by adenosine triphosphatase (ATPase) histochemistry combined with reactions against antifast and antislow myosin antibodies. The histochemical and immunohistochemical observations indicate a well-developed specialization of function within specific regions of the muscle. In parallel, analyses of native myosin isoforms and myosin heavy chain isoforms indicate two points. First, the histochemical "type IIe" fiber is predominant in cranial portions of the muscle, and myosin extracted from these regions exhibits a unique electrophoretic mobility not observed in the myosin isoforms of more traditional laboratory mammals. Second, the type I fibers are confined to the pectoralis abdominalis muscle and a small adjacent region of the caudal part of the pectoralis. This pattern of type I fiber distribution is considered a derived character state compared to muscle histochemical phenotype and isoform composition in the pectoralis muscles of other phyllostomids we have studied (Artibeus jamaicensis, Artibeus lituratus, Carollia perspicillata). We relate this to the unique locomotory needs of the common vampire bat, Desmodus rotundus. PMID- 8230236 TI - The vomeronasal duct has a protracted postnatal development in the mouse. AB - Recent evidence suggests that the accessory olfactory system (AOS) may mediate chemoreception before birth. Such a capability may allow the fetus to begin to sample chemical stimuli from the outside world, a possibility that has important developmental and evolutionary implications. Herein we describe the development in the mouse of the duct that connects the vomeronasal organ (VNO), containing the primary receptor neurons of the AOS, with the nasal cavity and thus with external stimuli. Twenty-four mice, four at each of six different ages from the last day of gestation through 25 days of age, were fixed and embedded in glycol methacrylate. Serial sections were examined under the light microscope so that the VNO duct could be reconstructed in three dimensions. Results confirm an earlier study which demonstrated that the VNO duct is not patent before birth. The duct becomes patent sometime after the first day of life but remains in an immature condition throughout the normal prepubertal period. During this period the duct is characterized by an internal surface that is rapidly desquamating such that the lumen of the duct contains sloughed tissue. These results suggest that the VNO is unlikely to function in the prenatal period, since the route for external stimuli to reach its receptor surface is blocked. The protracted period of VNO duct development reported here is consistent with the great bulk of data on the AOS of mammals which firmly establishes its role in the detection of pheromones which coordinate reproduction. PMID- 8230237 TI - Cephalometric correlates of echolocation in the chiroptera. AB - This study suggests that the evolution of head posture in bats is constrained by the demands of vocalization during echolocation. Nasal-emitting microchiropteran taxa are easily identified by their characteristic rotation of the basicranium ventrally about the cervical axis, the depression of the rostrum below the basicranial axis, and by the rotation of the lateral semicircular canals so as to maintain their horizontal orientation during flight. The converse is true for oral-emitting Microchiroptera. The general form of the microchiropteran skull has been canalized along two distinct evolutionary paths, respectively, towards oral emitting or nasal-emitting forms. PMID- 8230238 TI - In memoriam Dr George Rona 1924-1993. PMID- 8230239 TI - Negative lusitropy and abnormal calcium handling in hypoxic cardiac myocytes exposed to the calcium-sensitizer EMD 53998. AB - Positive inotropic agents that increase the sensitivity of myofilaments to calcium have recently been described (Kitada et al., 1987; Cottney et al., 1990; Ferroni et al., 1991; Lee and Allen, 1991; Beier et al., 1992). These drugs appear to augment contractility independently of cAMP or calcium, and thus may have fewer of the adverse side effects seen with other currently available agents (Katz, 1986; Packer 1989). The clinical utility of "calcium-sensitizers" has been questioned on the theoretical grounds that such agents may interfere with relaxation and impair diastolic function (Hajjar and Gwathmey, 1991). Previous studies have shown a small but significant negative lusitropic effect of the calcium sensitizer EMD 53998 in ferret papillary muscle, although this effect was considered to be outweighed by powerful augmentation of contractility. Modelling studies have suggested that the impairment of relaxation by calcium-sensitizers may be even more severe when myocardial calcium is abnormally elevated, such as in hypoxia (Allen and Orchard, 1987; Lodge and Gelband, 1988) and end-stage heart failure (Hajjar and Gwathmey, 1991). We have examined the effects of EMD 53998 and milrinone on contractility and calcium flux in a cell culture model of myocardial hypoxia. The results indicate that increased calcium sensitivity results in marked impairment of relaxation under hypoxic conditions, possibly due to the impaired calcium sequestration and increased calcium availability exhibited by hypoxic myocytes. These studies show that the effects of calcium sensitizers can be strongly influenced by the prevailing status of intracellular calcium handling, and may be deleterious in the diseased or ischemic myocardium. PMID- 8230240 TI - New occurrence of atrial natriuretic factor and storage in secretorially active granules in adult rat ventricular cardiomyocytes in long-term culture. AB - Under normal physiological conditions atrial natriuretic factor (ANF) in rat is stored in secretory granules in fetal and adult atrial cardiomyocytes (Cantin et al., 1984; de Bold, 1985), whereas in fetal and neonatal ventricular cardiomyocytes secretory granules are practically absent and ANF is directly secreted via the constitutive pathway (Bloch et al., 1986; Cantin et al., 1987). In ventricular adult rat cardiomyocytes however, ANF is down-regulated and only expressed in cells of interventricular septum (Reinecke, 1989). These ANF containing cells seem to constitute part of the Purkinje fibre system. As ventricular adult rat cardiomyocytes (ARC) in longterm culture in many ways re express the fetal program (Eppenberger et al., 1988; Eppenberger-Eberhardt et al., 1990), it was of interest to investigate whether this was also the case for ANF. Data in the present study show that immunoreactive (IR) ANF is upregulated in cultured ventricular ARC and is stored just as in cultured atrial ARC in "atrial-type" granules and secreted into the medium. A possible analogy with overload heart hypertrophy in vivo, where a reactivation of an early gene program including re-expression of ANF in ventricular myocytes has been described (Izumo et al., 1987; Chien et al., 1991), is proposed. PMID- 8230241 TI - The myocardial vasculature during ischemia and reperfusion: a target for injury and protection. AB - Ischemia and reperfusion may result in injury to one or more of the cellular components of the heart. In addition to damaging myocytes and their contractile capability, ischemia and reperfusion may inflict early and severe injury on the vasculature which, in turn, may further jeopardize the survival of the myocytes. While ischemia is known to cause progressive injury to endothelium and vascular smooth muscle it now appears that reperfusion can inflict additional, possibly severe, injury on the microcirculation which may compromise the return of normal coronary perfusion. This post-ischemic/reperfusion microvascular incompetence ranges from a transient exacerbation of the increase in vascular resistance initiated during ischemia, to a sustained loss of competent capillaries and eventually to the "no reflow" phenomenon which is characterized by the total inability of the affected tissue to be reperfused. Whereas "no reflow" may be of little importance if it occurs in already infarcted tissue, post-ischemic microvascular incompetence in potentially salvable myocardium could be of considerable significance. Evidence is presented that the vascular endothelium, and its ability to regulate coronary vascular tone, play a central and early role in the pathogenesis of myocardial injury. Mechanisms underlying microvascular injury have been identified and pharmacological strategies now exist for the effective manipulation of this injury--a prospect that is of considerable importance in the light of the widespread use of thrombolytic procedures for the reperfusion of the human myocardium. PMID- 8230242 TI - Effects of tetrandrine on Ca(2+)- and Na(+)-currents of single bullfrog cardiomyocytes. AB - Tetrandrine is a natural alkaloid classified as a calcium antagonist. However, its precise actions on Ca(2+)-currents in cardiac cells have not been fully characterized. In the present study, we have investigated the mechanism of action of tetrandrine on the Ca(2+)-currents of single bullfrog cardiac cells, using the patch-clamp technique. Tetrandrine slightly increased ICaL from negative holding potentials (-100 mV) at low concentrations (10 nM-1 microM) and inhibited it at higher concentrations. At depolarized holding potentials (-50 mV) only an enhanced inhibition was seen. Tetrandrine blockade of the L-type Ca(2+)-current (ICaL) was mostly tonic. This is similar to ICaL blockade by nifedipine but not by verapamil, the latter being mostly use-dependent. Use-dependent effects of tetrandrine and nifedipine were evident at high rates. Availability curves were shifted leftwards (10-12 mV) by tetrandrine (10 microM) and nifedipine (1 microM). The T-type Ca(2+)-current (ICaT), although less sensitive, was decreased by both agents in a voltage-independent way. Tetrandrine (10-30 microM) but not nifedipine (1-10 microM), depressed the Na(+)-current (INa) in tonic, use- and voltage-dependent manners. We conclude that tetrandrine and nifedipine share some common actions on cardiac Ca(2+)-channels, while showing differences in their actions on Na(+)-channels. The depression of INa by tetrandrine suggests it could be effective on supraventricular tachycardias. PMID- 8230243 TI - Pertussis toxin blocks adenosine A1 receptor mediated protection of the ischemic rat heart. AB - Adenosine and acetylcholine exert negative chronotropic and anti-adrenergic effects on nonischemic myocardium presumably via receptor coupling to the same or similar inhibitory guanine nucleotide binding protein (Gi). To determine whether the cardioprotective effect of adenosine is mediated via adenosine A1 receptor coupling to Gi proteins, isolated rat hearts, perfused at constant pressure and constant heart rate, were subjected to 30 min global normothermic (37 degrees C) ischemia and 45 min reperfusion. Untreated control hearts recovered 52 +/- 2% of preischemic left ventricular developed pressure (LVDP). Hearts treated for 10 minutes prior to ischemia with adenosine (100 microM) and the adenosine A1 receptor agonist cyclohexyladenosine (CHA, 0.25 microM) recovered 67 +/- 4% and 70 +/- 4%, respectively. Hearts treated with the non-specific muscarinic cholinergic agonist carbamylcholine (1 microM) exhibited similar enhanced postischemic recovery (70 +/- 3%). Pretreatment of rats with pertussis toxin (25 micrograms/kg i.p., 48 h prior to isolation) significantly reduced the negative chronotropic effects of adenosine and CHA. Pertussis toxin pretreatment also blocked the beneficial effects of adenosine (57 +/- 4% recovery) and CHA (49 +/- 4% recovery) on postischemic function. These results support the hypothesis that the salutary effect of adenosine on the ischemic myocardium is mediated via adenosine A1 receptor coupling to a pertussis toxin sensitive G protein, presumably Gi. PMID- 8230244 TI - Relationships between the effects of superoxide anion and palmitoyl-L-carnitine on the Ca(2+)-ATPase of vascular smooth muscle sarcoplasmic reticulum. AB - In order to understand interactions between oxidants and non-antioxidants, relationships between the effects of superoxide anion radical (O2-) and palmitoyl L-carnitine (PLC) on the Ca(2+)-ATPase of vascular smooth muscle sarcoplasmic reticulum were studied. PLC was ineffective in preventing the reduction of ferricytochrome c by O2-, suggesting that it has no O2(-)-scavenging properties. PLC exhibited a biphasic effect on Ca(2+)-ATPase activity, causing stimulation at low concentrations (3-10 microM) and inhibition at higher concentrations (30-1000 microM). The stimulatory effect on Ca(2+)-ATPase caused by 10 microM PLC was completely counteracted by O2-, whereas the stimulatory effect of L-cysteine (2 mM) was only diminished partially. Additive effects of PLC and L-cysteine attenuated the inhibitory effect of O2-, however, this appears to be due to L cysteine alone. These results demonstrate that O2- can inhibit stimulatory effects of PLC on the Ca(2+)-ATPase, however, PLC does not block the inhibitory effect of O2-. Understanding interactions between oxidants/antioxidants and non antioxidants at their cellular target proteins may provide essential information in the search for prevention of free radical injury. PMID- 8230245 TI - Preconditioning and post-ischaemic contractile dysfunction: the role of impaired oxygen delivery vs extracellular metabolite accumulation. AB - The aim of the present study was to identify components of ischaemia involved in the induction of preconditioning. Isolated rat hearts (n = 8 per group) were perfused with bicarbonate buffer. Following 10 min aerobic perfusion they were randomised and subjected to 5 min periods during which the perfusion conditions were: (i) normal aerobic perfusion (controls); (ii) zero flow ischaemia; (iii) low flow ischaemia (10% of control O2 delivery); (iv) hypoxia (10% of control O2 delivery); or (v) acidosis (pH 6.4). After these periods of "preconditioning", all hearts underwent 5 min aerobic perfusion followed by 40 min zero flow global ischaemia and 35 min reperfusion. Contractile function was measured at the beginning and at the end of the experiment. Despite profound differences in coronary flow during preconditioning, substantial and similar protection was observed in all groups preconditioned by transiently limiting oxygen delivery. Recovery of cardiac output was 66.7 +/- 6.3%, 58.7 +/- 5.1% and 62.6% +/- 3.3% in the zero flow, low flow and hypoxic groups, respectively, vs 31.0 +/- 3.0% in controls (all P < 0.05). In hearts subjected to acidosis there was no protection (recovery of cardiac output 38.1 +/- 2.7%). Impairment of oxygen delivery appears to be the principle component of ischaemia responsible for the induction of preconditioning. Metabolite accumulation appears to play no significant role. PMID- 8230246 TI - Reperfusion arrhythmias and purine wash-out in isolated rat and rabbit heart. Effect of allopurinol, dimethylthiourea and calcium reduction. AB - The effects of perfusate calcium reduction, allopurinol and dimethylthiourea on reperfusion-induced arrhythmias and purine wash-out in isolated rabbit and rat hearts were compared. The overall incidence of reperfusion-induced ventricular tachycardia (VT) was 88% and 94% and that of ventricular fibrillation (VF) was 44% and 88% in the control rabbit and rat hearts, respectively. VF was reduced to 10% and 0% in rat and rabbit hearts subjected to perfusate calcium reduction (0.4 mM for 1 min before ischemia and for 1 min before and throughout reperfusion), respectively. In allopurinol, 1 mM, perfused rat hearts the overall incidence of VF was not changed and only the incidence of a sustained VF (that lasting for at least 10 min) was reduced. VT and VF were prevented in allopurinol-perfused rabbit hearts. Dimethylthiourea, 10 mM, reduced the incidence of VF in rat hearts to 16% and did not significantly affect VT and VF in rabbit hearts. In untreated rat hearts, the major purine compounds washed out upon reperfusion were inosine, hypoxanthine, xanthine and urate. Allopurinol augmented the wash-out of adenosine and abolished that of xanthine and urate. In untreated rabbit hearts, the major purine washed out were inosine, adenosine and hypoxanthine. Allopurinol did not cause further increase in adenosine wash-out in rabbit hearts. We speculate that: (1) calcium mediated arrhythmogenic mechanism is operating both in reperfused rat and rabbit heart; (2) free radical mediated mechanism is of an importance only in rat heart; (3) neither a decreased free radical production secondary to xanthine oxidase inhibition nor the augmentation of adenosine wash-out is a likely explanation for the antiarrhythmic effect of allopurinol in reperfused hearts; and (4) high level of myocardial adenosine accumulation during ischemia, probably secondary to low xanthine oxidase activity, may play a role of a natural defence mechanism in ischemic/reperfused rabbit heart. PMID- 8230247 TI - Dobutamine prevents both myocardial stunning and phosphocreatine overshoot without affecting ATP level. AB - Catecholamines can overcome myocardial stunning. However, a previous report on energy metabolism in stunned myocardium during catecholamine infusion was based on the conventional biochemical methods which might affect contractile function. Twenty farm pigs were anesthetized and underwent 15 min coronary artery occlusion and 2 h reperfusion. Ten pigs were given 10 micrograms/kg/min dobutamine from immediately after and throughout the reperfusion (dobutamine group). The other ten pigs were given saline (control group). Phosphorus-31 magnetic resonance spectroscopy and sonomicrometry were done alternately. Dobutamine improved percent segment shortening after reperfusion (control/dobutamine = 3.8% 5.7%/11.7%-13.4%; P < 0.01). At 15 min ischemia, adenosine triphosphate (ATP) decreased (control/dobutamine = 72 +/- 8%/73 +/- 10%, n.s.), and remained depressed after reperfusion in both groups. After reperfusion, phosphocreatine (PCr) returned to and maintained the preischemic value in the dobutamine group, while in the control group, PCr overshoot (112 +/- 5%) was observed. Except for the presence and absence of PCr overshoot, there was no significant difference of ATP and PCr between the two groups, although rate pressure product was significantly higher in the dobutamine group than in the control group. Regional myocardial blood flow after reperfusion was significantly higher in the dobutamine group. Dobutamine may improve "stunning" through effective improvement of energy utilization and production, indicated by the disappearance of PCr overshoot and maintained ATP level. PMID- 8230248 TI - Effective population size, genetic diversity, and coalescence time in subdivided populations. AB - A formula for the effective population size for the finite island model of subdivided populations is derived. The formula indicates that the effective size can be substantially greater than the actual number of individuals in the entire population when the migration rate among subpopulations is small. It is shown that the mean nucleotide diversity, coalescence time, and heterozygosity for genes sampled from the entire population can be predicted fairly well from the theory for randomly mating populations if the effective population size for the finite island model is used. PMID- 8230249 TI - The complete intron/exon structure of Ephydatia mulleri fibrillar collagen gene suggests a mechanism for the evolution of an ancestral gene module. AB - We have completed the analysis of a genomic clone, G238, that contains most of the coding region of the sponge COLF1 fibrillar collagen gene. The main triple helical domain is encoded by 31 exons. Except for the 5' junction exon and the two last 3' exons (126 and 18 base pairs), all these exons are related to a 54-bp unit and begin with an intact glycine codon. A good correlation can be made between this sponge gene and a vertebrate fibrillar collagen gene, revealing the high conservation of the members of this family during evolution. The reconstitution of an ancestral collagen gene can be made by considering all the exon/intron junctions of these genes. We suggest that such an ancestral gene arose from multiple duplications of a 54-bp exon and a (54 + 45)-bp module. PMID- 8230250 TI - The paradox of MHC-DRB exon/intron evolution: alpha-helix and beta-sheet encoding regions diverge while hypervariable intronic simple repeats coevolve with beta sheet codons. AB - Twenty-one different caprine and 13 ovine MHC-DRB exon 2 sequences were determined including part of the adjacent introns containing simple repetitive (gt)n(ga)m elements. The positions for highly polymorphic DRB amino acids vary slightly among ungulates and other mammals. From man and mouse to ungulates the basic (gt)n(ga)m structure is fixed in evolution for 7 x 10(7) years whereas ample variations exist in the tandem (gt)n and (ga)m dinucleotides and especially their "degenerated" derivatives. Phylogenetic trees for the alpha-helices and beta-pleated sheets of the ungulate DRB sequences suggest different evolutionary histories. In hoofed animals as well as in humans DRB beta-sheet encoding sequences and adjacent intronic repeats can be assembled into virtually identical groups suggesting coevolution of noncoding as well as coding DNA. In contrast alpha-helices and C-terminal parts of the first DRB domain evolve distinctly. In the absence of a defined mechanism causing specific, site-directed mutations, double-recombination or gene-conversion-like events would readily explain this fact. The role of the intronic simple (gt)n(ga)m repeat is discussed with respect to these genetic exchange mechanisms during evolution. PMID- 8230251 TI - Chloroplast DNA codon use: evidence for selection at the psb A locus based on tRNA availability. AB - Codon use in the three sequenced chloroplast genomes (Marchantia, Oryza, and Nicotiana) is examined. The chloroplast has a bias in that codons NNA and NNT are favored over synonymous NNC and NNG codons. This appears to be a consequence of an overall high A + T content of the genome. This pattern of codon use is not followed by the psb A gene of all three genomes and other psb A sequences examined. In this gene, the codon use favors NNC over NNT for twofold degenerate amino acids. In each case the only tRNA coded by the genome is complementary to the NNC codon. This codon use is similar to the codon use by chloroplast genes examined from Chlamydomonas reinhardtii. Since psb A is the major translation product of the chloroplast, this suggests that selection is acting on the codon use of this gene to adapt codons to tRNA availability, as previously suggested for unicellular organisms. PMID- 8230252 TI - Endotherms, ectotherms, and mitochondrial genome-size variation. AB - The patterns of mitochondrial genome-size variation were investigated in endothermic and ectothermic species to examine the role that thermal habit might play in the evolution of animal mitochondrial DNA (mtDNA). Data on mtDNA size (the modal, largest, and smallest mtDNA reported within a species), the percent variation in mtDNA size (the difference in size between the largest and smallest mtDNAs divided by the model genome size for a given species), and the frequency of heteroplasmic individuals (those carrying more than one mtDNA length variant) were tabulated from the literature. Endotherms showed significantly less variation in mtDNA size and tended to have smaller mtDNAs than ectotherms. Further comparisons between endothermic and ectothermic vertebrates revealed that the largest genome and the percent variation in genome size were significantly smaller in the former than the latter. There was no difference between endotherms and ectotherms in the frequency of heteroplasmy. These data are discussed in light of two hypotheses: (1) more intense directional and purifying selection for small genome size in the cytoplasms of species with higher metabolic rates and (2) reduced mutation pressures generating mtDNA size variants in endotherms relative to those in ectotherms. The general trends are consistent with the selection hypothesis but in certain species mtDNA size variation appears to be governed by mutational pressures. To test these competing hypotheses further, comparative studies are proposed where mitochondrial genome size is quantified in sister taxa and tissue types with very different metabolic rates. PMID- 8230254 TI - Bryostatin 1: will the oceans provide a cancer cure? PMID- 8230253 TI - Molecular characterization and evolution of a duck mitochondrial genome. AB - We sequenced 6,478 bp of mitochondrial DNA from Peking duck (Anas platyrhyncos). Eight protein genes, 11 tRNAs, part of the small and large ribosomal subunits, and the control region sequences were compared to homologous chicken sequences. The gene organization in duck and chicken is identical but differs from other vertebrates in the juxtaposition of the tRNA(Glu)-ND6 genes next to the control region and in the lack of a hairpinlike structure between the genes for tRNA(Asn) and tRNA(Cys) used for light-strand replication. Protein, tRNA, and rRNA genes evolved mainly through base substitutions and small insertions and deletions. Transitions greatly outnumber transversions in the tRNA and rRNA genes, but this bias is not evident in protein genes; the control region has a higher proportion of transversions. The duck and chicken control regions show a high frequency of length mutations. Large A-T-rich nucleotide stretches dispersed across the region between the bidirectional transcription promoter and the heavy-strand replication origin in the chicken are absent in the duck. Sequence elements for heavy-strand replication in mammals are conserved in the duck and chicken control regions. Estimates of divergence for ribosomal RNAs and proteins based on total substitutions, transversions, and amino acid replacements show that all the duck/chicken values are lower than the corresponding mammal/mammal (cow, human, mouse) values. If paleontological data suggesting that avian and eutherian ordinal radiation occurred at approximately the same time are correct, this suggests that at great evolutionary distance, rate of mitochondrial DNA evolution in birds is somewhat decelerated compared to mammals. PMID- 8230255 TI - Board recommends changes to draft breast cancer screening guidelines. PMID- 8230256 TI - Researchers engineering new cancer drug delivery systems. PMID- 8230257 TI - Gene-marking studies in bone marrow may yield treatments. PMID- 8230258 TI - Prevention trial for prostate cancer piques public interest. PMID- 8230259 TI - Disease prevention research gains ground. PMID- 8230260 TI - Cancer prevention and control within the National Cancer Institute's clinical trials network: lessons from the Community Clinical Oncology Program. PMID- 8230261 TI - Phase I study of bryostatin 1: assessment of interleukin 6 and tumor necrosis factor alpha induction in vivo. The Cancer Research Campaign Phase I Committee. AB - BACKGROUND: Many oncogenes have been shown to code for growth factor receptors that are involved in regulation of cell growth and proliferation and can activate transcription via protein kinase C. Bryostatin 1, a partial agonist of protein kinase C, has demonstrated potent antitumor activity in vitro and in vivo in human tumor xenografts. PURPOSE: The aim of this phase I study was to determine the optimal dosage and toxicity profile of bryostatin 1 and its influence on cytokine release in vivo. METHODS: Three successive cohorts consisting of 35 patients with various malignant tumors were treated with bryostatin 1 by intravenous infusion over 1 hour as follows: cohort A--35 micrograms/m2 (three patients) or 50 micrograms/m2 (eight patients) once every 2 weeks; cohort B--25 micrograms/m2 once a week (eight patients); and cohort C--25 micrograms/m2 once a week for 3 weeks, with no treatment during the 4th week (16 patients). Plasma levels of tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) were measured by immunoradiometric assay and by radioimmunoassay, respectively. RESULTS: The dose-limiting toxicity was grade 3 or 4 myalgia in four of 11 patients in cohort A, in two of eight in cohort B, and in none of 16 in cohort C. Occurrence of myalgia was dose related. There was no significant myelosuppression, apart from a small and transient fall in platelet count. Six patients experienced acute but transient skin flushing, dyspnea, hypotension, and bradycardia, probably related to the bryostatin 1 vehicle. TNF-alpha and IL-6 were detected in plasma at 2 and 24 hours after treatment, respectively, and the levels were dose related (P = .02). Two patients with metastatic malignant melanoma had partial remission after three or four cycles of therapy; remission lasted 6 weeks and 10+ months, respectively. CONCLUSIONS: The dose-limiting toxicity of bryostatin 1 was myalgia. Plasma IL-6 and TNF-alpha concentrations were increased within 24 hours of therapy. Antitumor activity against malignant melanoma was observed early in the course of treatment. IMPLICATIONS: The recommended dosage of bryostatin 1 for phase II studies is 25 micrograms/m2 by intravenous infusion for 1 hour once a week for 3 weeks, with no treatment in the 4th week. IL-6 and TNF-alpha plasma concentrations may be useful in monitoring biological activity of bryostatin 1. Future studies should explore use of this drug with other conventional immune modulators and conventional cytotoxic drugs. PMID- 8230262 TI - Migration patterns and breast cancer risk in Asian-American women. AB - BACKGROUND: Breast cancer incidence rates have historically been 4-7 times higher in the United States than in China or Japan, although the reasons remain elusive. When Chinese, Japanese, or Filipino women migrate to the United States, breast cancer risk rises over several generations and approaches that among U.S. Whites. PURPOSE: Our objective was to quantify breast cancer risks associated with the various migration patterns of Asian-American women. METHODS: A population-based, case-control study of breast cancer among women of Chinese, Japanese, and Filipino ethnicities, aged 20-55 years, was conducted during 1983-1987 in San Francisco-Oakland, California, Los Angeles, California, and Oahu, Hawaii. We successfully interviewed 597 case subjects (70% of those eligible) and 966 control subjects (75%). RESULTS: A sixfold gradient in breast cancer risk by migration patterns was observed. Asian-American women born in the West had a breast cancer risk 60% higher than Asian-American women born in the East. Among those born in the West, risk was determined by whether their grandparents, especially grandmothers, were born in the East or the West. Asian-American women with three or four grandparents born in the West had a risk 50% higher than those with all grandparents born in the East. Among the Asian-American women born in the East, breast cancer risk was determined by whether their communities prior to migration were rural or urban and by the number of years subsequently lived in the West. Migrants from urban areas had a risk 30% higher than migrants from rural areas. Migrants who had lived in the West for a decade or longer had a risk 80% higher than more recent migrants. Risk was unrelated to age at migration for women migrating at ages less than 36 years. Ethnic-specific incidence rates of breast cancer in the migrating generation were clearly elevated above those in the countries of origin, while rates in Asian-Americans born in the West approximated the U.S. White rate. CONCLUSIONS: Exposure to Western lifestyles had a substantial impact on breast cancer risk in Asian migrants to the United States during their lifetime. There was no direct evidence of an especially susceptible period, during either menarche or early reproductive life. IMPLICATIONS: Because heterogeneity in breast cancer risk in these ethnic populations is similar to that in international comparisons and because analytic epidemiologic studies offer the opportunity to disentangle correlated exposures, this study should provide new insights into the etiology of breast cancer. PMID- 8230263 TI - Quantitative study of monocyte chemoattractant protein-1 (MCP-1) in cerebrospinal fluid and cyst fluid from patients with malignant glioma. AB - BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1) is a 76-amino acid protein that attracts monocytes. In vitro studies have reported high levels of MCP-1 messenger RNA expression, as well as the presence of MCP-1, in malignant glioma cells. PURPOSE: Our purpose was to determine whether an MCP-1 assay could be used in a clinical setting 1) to differentiate malignant from benign gliomas and from nontumor disorders of the central nervous system and 2) to detect subarachnoid dissemination of glioma cells. METHODS: MCP-1 levels in cerebrospinal fluid (CSF) and cyst fluid were measured with a sandwich enzyme-linked immunosorbent assay (ELISA) that we had previously developed. We measured MCP-1 levels in CSF samples from 19 patients with malignant glioma (glioblastoma, 10; anaplastic astrocytoma, six; anaplastic oligodendroglioma, two; and ependymoblastoma, one), nine patients with benign glioma, and seven patients with nontumor disorders of the central nervous system. Cyst fluids from four patients with malignant glioma (anaplastic astrocytoma) were also tested. The correlation between MCP-1 concentration in the CSF and subarachnoid dissemination of malignant glioma cells was also studied. RESULTS: The MCP-1 concentration (mean +/- SE) in CSF samples from patients with malignant glioma (2.3 +/- 0.4 ng/mL) was significantly higher than that from patients with benign glioma (0.6 +/- 0.1 ng/mL) (P < .01) or from patients with no tumor (0.5 +/- 0.1 ng/mL) (P < .01). Furthermore, CSF samples from patients with subarachnoid dissemination of malignant glioma contained significantly higher amounts of MCP-1 than those from patients without dissemination (P < .05). Cyst fluids from four of the patients with malignant glioma contained high concentrations of MCP-1. CONCLUSIONS: These results indicate that MCP-1 is produced by malignant glioma in vivo as well as in vitro and suggest that testing for MCP-1 in CSF may be useful in the clinic to differentiate malignant glioma from benign glioma and to detect subarachnoid dissemination of the tumor cells. IMPLICATIONS: The MCP-1 ELISA in CSF may lead to more accurate diagnosis of malignant glioma and detection of subarachnoid dissemination of tumor cells, facilitating selection of patients with these conditions for appropriate therapy. PMID- 8230264 TI - Preoperative and postoperative combination chemotherapy for potentially resectable gastric carcinoma. AB - BACKGROUND: Median survival of patients with local-regional gastric carcinoma is 10 months. Resection of the primary tumor and regional lymph nodes, with tumor free margins (curative resection), has been the most effective treatment for local-regional gastric carcinoma. However, median survival of patients with curative resection of gastric carcinoma is 24 months, and the 5-year survival rate is about 20%. A single institution pilot study has established the feasibility of administering two courses of chemotherapy preoperatively and three courses postoperatively. In another study, a 15% pathologically documented complete response (pathologic complete response) has been reported in unresectable gastric carcinoma treated with etoposide, doxorubicin, and cisplatin. PURPOSE: Our purpose was to increase the curative resection rate in potentially resectable gastric carcinoma and to delay or eliminate micrometastases and thus improve survival. We also evaluated clinical and pathologic response to chemotherapy. METHODS: Forty-eight previously untreated patients with potentially resectable gastric carcinoma received a chemotherapy regimen (EAP) consisting of etoposide (120 mg/m2 intravenously over a 2-hour period on days 4, 5, and 6), doxorubicin (20 mg/m2 as a 10-minute intravenous infusion on days 1 and 7), and cisplatin (40 mg/m2 as a 1-hour intravenous infusion on days 2 and 8). Patients received three courses of chemotherapy before resection, and responding patients received two courses postoperatively. Clinical and pathologic response rates, toxicity, patterns of treatment failure, and survival times were assessed. RESULTS: A median of three courses (range, 1-5) of preoperative therapy was administered; six (12%) of the 48 patients had clinical complete response, and nine (19%) had partial response. Forty-one (85%) underwent surgery; 37 (90%) of these 41 (77% of the 48 patients) had a curative resection. There were no pathologic complete responses. Median survival for all patients is 15.5 months (range, 2-29+ months). Therapy was discontinued because of the toxic effects in one patient before surgery and in six patients after surgery. Doses were reduced in 37 patients (77%), mainly because of hematologic toxicity. Nineteen (40%) were hospitalized because of toxic effects, including 15 patients who developed fever with neutropenia. Grade 3 or 4 nausea and vomiting occurred in 15 patients and grade 3 or 4 diarrhea in seven patients. One death was directly related to chemotherapy. CONCLUSIONS: These data support that administration of preoperative and postoperative chemotherapy for local-regional gastric carcinoma is feasible in a multi-institutional setting. Our findings demonstrate that this EAP regimen is modestly active but is associated with substantial toxicity. IMPLICATIONS: Use of preoperative chemotherapy in resectable gastric carcinoma merits further evaluation, but more effective drug regimens will be required before a controlled trial is initiated. PMID- 8230265 TI - Immune response induced in small-cell lung cancer by maintenance therapy with interferon gamma. AB - BACKGROUND: Chemotherapy, with or without radiotherapy, results in a 30%-40% complete response rate in small-cell lung cancer (SCLC), but approximately 90% of patients who have complete remission die within 2 years after relapse with chemoresistant disease. Randomized clinical studies of maintenance chemotherapy after complete response have failed to demonstrate survival advantage. However, studies have shown that the human cytokine interferon gamma (IFN-gamma) induces immune response in humans, including T-cell activation and expression of class II major histocompatibility complex (HLA-DR) and receptor for the Fc portion of immunoglobulin on monocytes. It has also been demonstrated that recombinant IFN gamma (rIFN-gamma) induces immunomodulation and has antiproliferative activity. PURPOSE: In vivo effects of rIFN-gamma treatment were characterized by flow cytometric analysis of peripheral blood mononuclear cells in patients with SCLC who received rIFN-gamma as maintenance treatment. METHODS: After induction chemotherapy and radiotherapy, 100 patients who achieved a complete remission were randomly assigned to receive rIFN-gamma at a dose of 0.2 mg (4 x 10(6) units) once a day, subcutaneously, for 6 months, or observation only. In 31 patients, peripheral mononuclear cells were obtained prior to the study and at weeks 4, 8, and 12 for serial monitoring of immune response. By flow cytometric analysis, we identified the lymphocyte and monocyte populations using characteristic differences in electronic volume and right-angle scatter. In these populations, we determined the mean fluorescence channel after staining for CD14 (antigen expressed on monocytes), CD3 (antigen expressed on T lymphocytes), and HLA-DR (HLA class II expressed by monocytes and activated lymphocytes). To determine the number of Fc receptors per cell, an Fc receptor assay was performed using the monocyte cell line U937 as a standard. RESULTS: At weeks 4, 8, and 12, expression of HLA-DR and Fc receptors on monocytes in patients who received rIFN gamma was significantly higher than that in untreated patients, and the difference was statistically significant. The number of Fc receptors per monocyte consistently increased during the rIFN-gamma treatment and reached a fivefold elevation at week 12. There was no statistically significant difference in lymphocyte surface antigen expression between the treated and untreated groups. CONCLUSION: The dose of rIFN-gamma used in this study resulted in immune stimulation in patients with SCLC who had complete remission after induction therapy. The in vivo immunomodulatory activity of rIFN-gamma in such patients is characterized by a strong monocyte activation but no significant alteration in T cell activation. PMID- 8230266 TI - Descriptive clinicopathologic study of 17 patients with endometrial cancer during or after adjuvant tamoxifen in early breast cancer. AB - BACKGROUND: Studies have shown that patients with early-stage endometrial cancer who have previously used endogenous estrogen (oral contraceptives or estrogen replacement therapy) have a favorable prognosis. This has not yet been demonstrated for patients with early-stage endometrial cancer who have received tamoxifen. In addition, studies have raised the question of whether women receiving tamoxifen are at increased risk of endometrial cancer. PURPOSE: Our aim was to determine whether the prognosis is favorable for patients with diagnosis of endometrial cancer after adjuvant treatment with tamoxifen for breast cancer. METHODS: We matched 931 patients from the Stockholm Adjuvant Tamoxifen Trial in early breast cancer against the Swedish Cancer Registry and identified 17 who subsequently had endometrial cancer. These patients had been randomly assigned to receive 40 mg/d tamoxifen orally for 2 years beginning 4 weeks after surgery for breast cancer. Histologic specimens, patient records, and death certificates were reviewed to verify treatment and causes of death. RESULTS: Thirteen of the 17 patients diagnosed with endometrial cancer were alive; for three of the four who had died, the cause of death was endometrial cancer. All 16 evaluable tumors except one were World Health Organization (WHO) histologic grades I-II. Only one patient had advanced disease (stage IV); the remaining tumor was a mixed mesodermal malignant tumor that could not be classified under the WHO grading system. Median time for adjuvant tamoxifen use was 24 months (range, 6-60 months) with a median cumulative tamoxifen dose of 29 g (range, 7-72 g). Median time from initiation of adjuvant tamoxifen to diagnosis of endometrial cancer was 32 months (range, 6-130 months). Ten-year actuarial survival after diagnosis of endometrial cancer for the 17 patients treated with tamoxifen was 73%. CONCLUSION: Because of the small number of patients, our results do not rule out the possibility of a favorable prognosis for patients with a diagnosis of endometrial cancer following tamoxifen treatment. IMPLICATIONS: The incidence of secondary endometrial cancer reported in this study following treatment of breast cancer patients with tamoxifen at doses of 40 mg/d in a large clinical trial is higher than that reported for previous large trials of tamoxifen at doses of 20 mg/d. Thus, tamoxifen dosage may be a critical factor in the subsequent occurrence of endometrial cancer. Our results also suggest two important considerations for improved follow-up in long-term tamoxifen trials: careful registration of second cancers and routine gynecologic examinations to ensure early detection of endometrial cancer. PMID- 8230267 TI - Secondary myelodysplasia and acute leukemia following carboplatin-containing combination chemotherapy for ovarian cancer. PMID- 8230268 TI - Use of heparin as a topical antithrombotic and antiinflammatory agent. PMID- 8230269 TI - Lack of carcinogenicity in mice exposed once to high concentrations of 1,3 butadiene. PMID- 8230270 TI - Advice to doctors. PMID- 8230271 TI - Human papillomavirus and cervical intraepithelial neoplasia. PMID- 8230272 TI - Chemoprevention studies: controlling effects of initial nutrient levels. PMID- 8230273 TI - Increasing incidence of primary central nervous system tumors in the elderly: real increase or improved detection? PMID- 8230274 TI - Lung cancer: another consequence of a high-fat diet? PMID- 8230275 TI - Elevated estrogen 16 alpha-hydroxylase activity: is this a genotoxic or nongenotoxic biomarker in human breast cancer risk? PMID- 8230276 TI - Spontaneous cancer remissions spark questions. PMID- 8230278 TI - Presidential commission seeks greater funding for breast cancer research. PMID- 8230277 TI - Differentiation agents yield treatment, prevention options. PMID- 8230279 TI - Major cancer screening trial launched in 10 U.S. centers. PMID- 8230280 TI - Saturated fat intake and lung cancer risk among nonsmoking women in Missouri. AB - BACKGROUND: Although the vast majority of lung cancer cases in women are caused by smoking, 9%-20% of cases occur in nonsmokers. Previous epidemiologic research on the relationship between lung cancer and diet has shown that fruit and vegetable consumption may confer a protective effect against lung cancer, while a diet rich in cholesterol and fat may increase risk. PURPOSE: The purpose of this case-control study was to examine the effects of a broad range of dietary factors on the risk of lung cancer in a population of nonsmoking white women 30-84 years of age. METHODS: A telephone-administered questionnaire was used to determine and/or verify eligibility with regard to age, gender, race, and smoking status. In a second interview at the participant's home, a widely used food frequency questionnaire was filled out, and logistic regression was subsequently used to analyze the responses. We obtained dietary information on 429 case subjects who had a diagnosis of lung cancer reported to the Missouri Cancer Registry between June 1, 1986, and June 1, 1991, and 1021 control subjects. If a case subject had died or was too ill to be interviewed, next-of-kin familiar with the woman's diet were interviewed instead. Of the 429 women with lung cancer, 211 (49%) had lung adenocarcinoma. RESULTS: A strongly increasing trend in lung cancer risk was observed with increased saturated fat consumption among these non-smoking women; the relative risk was more than sixfold greater for the highest quintile of consumption than for the lowest quintile. The effect of saturated fat was more pronounced for adenocarcinoma than for other cell types. Weekly servings of beans and peas were significantly related to decreased lung cancer risk, while citrus fruit and juice showed a twofold increase in risk; this trend was also significant. CONCLUSION: By focusing on non-smoking women with lung cancer, including a large number with adenocarcinoma, we observed a clear association with saturated fat consumption that may have been masked in earlier studies of lung cancer involving a high percentage of smokers. PMID- 8230281 TI - Upregulation of estradiol C16 alpha-hydroxylation in human breast tissue: a potential biomarker of breast cancer risk. AB - BACKGROUND: The biotransformation of the natural estrogen 17 beta-estradiol (E2) via the C16 alpha-hydroxylation pathway is elevated in patients with breast cancer, in subjects at increased risk for developing breast cancer, and in c-Ha ras-initiated mouse mammary epithelial cells. PURPOSE: To determine whether differences in the extent of E2 C16 alpha-hydroxylation are related to the risk of developing breast cancer, we examined the extent of biotransformation of E2 via the C16 alpha-hydroxylation pathway in the mammary terminal duct lobular units (TDLUs), epithelial organoids that are a presumptive target site of human breast carcinogenesis, and in nontarget component mammary fat tissue. METHODS: Noninvolved mammary tissue was obtained from four patients undergoing reduction mammoplasty and from four undergoing mastectomy for breast cancer. A radiometric assay that measures 3H2O formation caused by stoichiometric 3H exchange from [C16 alpha-3H]E2 was utilized to compare the relative extent of C16 alpha hydroxylation in explant cultures of TDLUs and mammary fat. RESULTS: The extent of E2 C16 alpha-hydroxylation was 1.83-fold higher (95% confidence interval [CI] = 1.71-1.97) in the TDLUs from reduction mammoplasty (i.e., "low-risk") patients and 7.96-fold higher (95% CI = 6.38-10.55) in the TDLUs from mastectomy (i.e., "high-risk") patients than in the corresponding values observed in the mammary fat. In the TDLUs obtained from the patients undergoing mastectomy for cancer, the extent of this metabolism was 4.56-fold higher (95% CI = 3.97-5.33) than that observed in TDLUs obtained from reduction mammoplasty patients who did not have cancer. CONCLUSION: The increase in the extent of C16 alpha-hydroxylation of E2 in the epithelial organoids of the human breast, TDLUs in particular, may be an important factor for breast cancer induction. This upregulation may represent an endocrine biomarker for the risk of developing breast cancer. IMPLICATION: A larger prospective study is required to confirm the clinical significance of this endocrine biomarker. PMID- 8230282 TI - Constitutive variability in the pharmacokinetics of the natural retinoid, all trans-retinoic acid, and its modulation by ketoconazole. AB - BACKGROUND: All-trans-retinoic acid (all-trans RA) induces complete remission in most patients with acute promyelocytic leukemia (APL). However, continuous oral dosing results in progressive decline in plasma drug concentrations, which is associated with relapse and resistance to this retinoid. We speculated that the decline in drug levels, indicating acquired resistance, resulted partly from inducible cytochrome-P450 oxidative enzymes, which can catabolize all-trans RA. PURPOSE: We studied the clinical pharmacology of all-trans RA in cancer patients to determine possible mechanisms of acquired resistance and evaluated the potential for reversal by ketoconazole, an inhibitor of cytochrome-P450 oxidative enzymes. METHODS: Serial plasma samples were obtained from 54 patients with APL or advanced lung cancer after a single oral dose of all-trans RA (45 mg/m2). In the 34 patients with advanced lung cancer, all-trans RA (45 mg/m2) was administered twice daily for 4 weeks, and, on days 2, 28, and 29, serial plasma samples were again obtained after a single 45-mg/m2 dose. One hour prior to drug administration on days 2 and 29, a single oral dose (200-1200 mg) of ketoconazole was administered. Endogenous plasma concentrations of all-trans RA and 13-cis retinoic acid were measured in a subset of these patients and in 11 with early stage lung cancer. RESULTS: The mean area under the curve for plasma drug concentration times time (AUC) for all-trans RA on day 1 varied substantially among patients. Compared with patients with APL, the 28 patients with advanced lung cancer who completed therapy demonstrated significantly lower AUC levels on day 1 (P = .06); a subgroup with levels less than 300 ng/mL per hour on day 1 had lower endogenous plasma all-trans RA concentrations than patients with APL or early-stage lung cancer or 14 normal subjects. Following continuous oral treatment, the mean day 28 AUC for all-trans RA was significantly lower than that on day 1 (213 ng/mL per hour versus 467 ng/mL per hour; P < .01), a decline significantly attenuated by ketoconazole, which increased the mean plasma all trans RA AUC on day 29 to 375 ng/mL per hour (P < .01). CONCLUSION: Reported variability for the pharmacokinetics of all-trans RA may result from disease related or population-based differences in basal catabolic rates influenced by genetic or environmental factors. However, the pattern of inducible catabolism of all-trans RA is not disease specific. Ketoconazole attenuates this accelerated catabolism, suggesting that oxidation by cytochrome-P450 enzymes is an important pathway for both constitutive and induced pathways of all-trans RA metabolism. PMID- 8230283 TI - Circadian dependence of interferon antitumor activity in mice. AB - BACKGROUND: Chronobiological studies with anticancer drugs have shown that their effectiveness and/or toxicity is significantly influenced by the time of their administration in the circadian cycle. Previous studies also have shown that the myelotoxicity of interferons is similarly influenced. PURPOSE: This study was undertaken to evaluate the antitumor activity of interferons as a function of their administration to animals at defined points in the circadian cycle with equal light and dark periods. METHODS: A murine tumor model was employed. Following adaptation to alternating cycles of 12 hours of light and 12 hours of dark for a period of 2-3 weeks, C57BL/6 mice were inoculated with B16 melanoma cells intraperitoneally at different hours after light onset. Exactly 24 hours after inoculation, each group received intraperitoneal injections of either recombinant human interferon alpha (rHuIFN-alpha A/D), recombinant murine IFN gamma (rMuIFN-gamma), or interferon-carrier solution as control (once a day for 5 days) and were monitored for the length of their survival. RESULTS: The antitumor activity (calculated as percent increased life span) of both rHuIFN-alpha A/D and rMuIFN-gamma varied with the points at which they were administered in the circadian cycle. However, the points showing minimum and maximum activity for rHuIFN-alpha A/D (12-16 and 0-4 hours after light onset, respectively) did not correspond with the points for the rMuIFN-gamma (0-8 and 16 hours after light onset, respectively). To generate maximum antitumor activity, approximately fivefold higher amounts of rHuIFN-alpha A/D were required at 12 than at 4 hours after light onset (dose range, 3333-90,000 IU/d) (P < .0001). Similarly, for rMuIFN-gamma at least 8.5-fold greater amounts were required at 8 than at 16 hours after light onset (dose range, 667-6000 IU/d) (P < .01). CONCLUSIONS: In the murine tumor model, administration of rHuIFN-alpha A/D at 4 hours after light onset and rMuIFN-gamma at 16 hours after light onset may produce maximum antitumor activity. PMID- 8230284 TI - Second cancers among long-term survivors of non-Hodgkin's lymphoma. AB - BACKGROUND: Patients with non-Hodgkin's lymphoma (NHL) are at increased risk for second cancers. Few studies, however, include long-term survivors, and none report risk for second cancer among NHL patients surviving 15 or more years. PURPOSE: Our aim was to examine the pattern of second cancers among long-term survivors of NHL. METHODS: A cohort of 6171 patients diagnosed with NHL as a first primary cancer and who survived 2 or more years was identified within population-based tumor registries in Sweden, Ontario, and Iowa and within the affiliated tumor registry of The Netherlands Cancer Institute. Nearly 1000 NHL patients lived 15 or more years after diagnosis. Tumor registry files were searched for new invasive primary malignancies. RESULTS: Second cancers were reported in 541 subjects (observed-to-expected ratio [O/E] = 1.37; 95% confidence interval = 1.26-1.49), with significant excesses seen for all solid tumors (O/E = 1.28), acute nonlymphocytic leukemia (O/E = 4.83), melanoma (O/E = 2.38), Hodgkin's disease (O/E = 12.0), and cancers of the lung (O/E = 1.36), brain (O/E = 2.33), kidney (O/E = 2.07), and bladder (O/E = 1.77). Among 15-year survivors, significantly increased risks persisted for all second cancers (O/E = 1.45), solid tumors (O/E = 1.37), bladder cancer (O/E = 3.24), and Hodgkin's disease (O/E = 25.0). The actuarial risk of developing a second cancer 3-20 years after diagnosis of NHL was 21%, compared with a population expected cumulative risk of 15%. CONCLUSIONS: Patients with NHL continue to be at significantly elevated risk of second primary cancer for up to two decades following diagnosis. The pattern of risk suggests the influence of treatment as well as factors associated with the underlying disease. IMPLICATIONS: Quantitative studies of second cancer following NHL are needed to clarify the role of antecedent therapy, shared risk factors, host susceptibility, and other etiologic and diagnostic influences. Despite the generally advanced age of patients with NHL, the persistently elevated risk of second cancers should alert clinicians to the importance of continued medical surveillance. PMID- 8230285 TI - Synergism and lack of cross-resistance between short-term and continuous exposure to fluorouracil in human colon adenocarcinoma cells. AB - BACKGROUND: Our recent findings in vitro in the human colon adenocarcinoma cell line HCT-8 suggest that resistance to fluorouracil (5-FU) in patients with advanced colorectal cancer might be overcome by use of a different treatment schedule. PURPOSE: We tested the hypothesis that HCT-8 cells resistant to short term 5-FU exposure retain sensitivity to continuous exposure and studied interactions between the two schedules. METHODS: HCT-8 cell lines resistant to short-term (pulse) treatment with 5-FU or to continuous exposure were obtained by six exposures to different concentrations of 5-FU for 4 hours or 7 days. We used a monolayer clonogenic assay to determine 5-FU-induced cell kill in resistant HCT 8 cells and sensitive parent cells. Parent cells were exposed to different concentrations of 5-FU for 1, 4, or 24 hours (short term), for 7 days (continuous exposure), or in a combination of both types of schedules. In a study of the mechanism of interaction between short-term and continuous exposure in parent cells, we performed flow cytometric DNA analysis to determine the percentage of cells in S phase and assays of thymidylate synthase inhibition in intact cells and of incorporation of [6-3H)]5-FU nucleotides into nucleic acids. RESULTS: Sensitive HCT-8 cells became fully resistant to 5-FU within five or six treatments, and low-dose continuous exposure almost immediately produced resistant clones. HCT-8 cells resistant to 5-FU given every 4 hours retained full sensitivity to continuous exposure, suggesting lack of cross-resistance between the two schedules, but cells resistant to continuous exposure were cross resistant to short-term treatment. Parent cells showed a statistically significant (synergistic) enhancement of the cytotoxic activity for 5-FU exposure for 1 hour (100, 300, or 500 microM) followed by continuous exposure (0.5, 1, or 2 microM) or 4 hours (10, 30, or 60 microM) followed by continuous exposure (1 or 2 microM). Short-term plus continuous exposure produced a marked increase in percentage of S-phase cells, compared with the percentage for each schedule alone. The combination of 1-hour exposure and continuous exposure (1000 and 2 microM, respectively) produced a marked accumulation of cells in S phase at 24 hours (59%), which lasted up to 96 hours (53%). The combination of the two schedules produced only additive enhancement of thymidylate synthase inhibition as well as incorporation of [6-3H]5-FU nucleotides into nucleic acids of HCT-8 cells. CONCLUSIONS: Our findings provide a rationale for the use of bolus 5-FU and continuous infusion 5-FU in sequence. IMPLICATION: We are conducting a clinical trial of bolus methotrexate followed by continuous-infusion 5-FU plus leucovorin. PMID- 8230286 TI - Assuring access to state-of-the-art care for U.S. minority populations: the first 2 years of the Minority-Based Community Clinical Oncology Program. AB - BACKGROUND: The Minority-Based Community Clinical Oncology Program (MBCCOP) was initiated in September 1990 to expand the National Cancer Institute's (NCI's) clinical trials network to minority populations. Institutions, organizations, and/or physician groups that had more than 50% of new cancer patients from minority groups were eligible to participate. There has been no previous evaluation of the MBCCOP. PURPOSE: This study was designed to describe the early implementation of the MBCCOP and identify the challenges that have emerged in developing a network aimed at increasing the participation of minority populations in clinical trials. METHODS: Data were taken from primary and secondary sources, including site visits and patient log data, that described performance of 12 MBCCOP centers initially funded in September 1990. Accrual was measured by the number of credits earned per MBCCOP for patients enrolled in research protocols for cancer treatment or for prevention and control, which includes activities such as early detection, pain control, and rehabilitation. These accrual credits, assigned by the NCI, were based on the complexity of the protocol and the amount of resources expected to be required for accrual of patients by the MBCCOP. RESULTS: Data for the first 2 years of the MBCCOP showed that 344 patients were accrued to trials of treatment protocols from June 1, 1990, to May 31, 1991, and this number increased to 470 during the second accrual year, June 1, 1991, to May 31, 1992. Similarly, accrual of patients to cancer prevention and control studies increased from 256 in 1990-1991 to 423 in 1991 1992. More than 70% of the MBCCOP patients entered in studies were from minority populations. The proportion of eligible MBCCOP patients entered into treatment protocols was identical with that experienced by the initial Community Clinical Oncology Program (CCOP). Results also demonstrated that MBCCOP centers operate in an environment characterized by socio-economic decline and limited resources, both having substantial effects on the implementation of clinical trials among minorities. While minority patients are willing to participate in clinical trials, there are profound barriers involving language, logistics, and the appropriateness of available protocols. Participating physicians, nurses, and support personnel report a high level of agreement with program goals and have developed unique approaches to meeting the challenges faced in the implementation of this program. CONCLUSIONS: The MBCCOPs have demonstrated their ability to participate in clinical trials. Evaluation reveals, however, that they are emerging organizations influenced by factors endemic to the community they serve and their own structure. The MBCCOPs are confronting substantial challenges, yet they provide an important link to the overall NCI clinical trials network. PMID- 8230287 TI - Detection of chromosomal polysomy in oral leukoplakia, a premalignant lesion. PMID- 8230288 TI - Intrinsic radiation sensitivity: no correlation with the metastatic potential of human and murine tumor cell lines. PMID- 8230289 TI - High-dose chemotherapy with autologous bone marrow support in node-positive melanoma. PMID- 8230290 TI - Skeptics of oral administration of shark cartilage. PMID- 8230291 TI - Selenium and DDE in breast fat of breast cancer patients: their relationship to hormone receptors in breast tissue. PMID- 8230292 TI - Sigmoidoscopy. PMID- 8230293 TI - Evaluation of histone sequence and modifications by electrospray ionization mass spectrometry and tandem mass spectrometry. PMID- 8230294 TI - Electrospray ionization mass spectrometric characterization of acrylamide adducts to hemoglobin. AB - The most common procedure to identify hemoglobin adducts has been to cleave the adducts from the protein and characterize the adducting species, by, for example, derivatization and gas chromatography/mass spectrometry. To extend these approaches we used electrospray ionization mass spectrometry (ESI-MS) to characterize adducted hemoglobin. For this we incubated [14C]acrylamide with the purified human hemoglobin (type A0) under conditions that yielded high adduct levels. When the hemoglobin was separated by reversed-phase high-performance liquid chromatography (HPLC), 65% of the radioactivity copurified with the beta subunit. Three adducted species were prominent in the ESI mass spectrum of the intact beta-subunit, indicating acrylamide adduction (i.e., mass increase of 71 Da) and two additional unidentified moieties with mass increments of 102 and 135 Da. Endoproteinase Glu-C digestion of the adducted beta-subunit resulted in a peptide mixture that, upon reversed-phase HPLC separation, provided several radiolabeled peptides. Using ESI-MS we identified these as the V91-101 and V102 122 peptides that represent the cysteine-containing peptides of the beta-subunit. These results provide definitive information on acrylamide-modified human hemoglobin and demonstrate that ESI-MS provides valuable structural information on chemically adducted proteins. PMID- 8230295 TI - Measurement of DNA adducts using surface-enhanced Raman spectroscopy. AB - Hazardous pollutants emitted from energy-related technologies, chemical industries, or waste materials are of increasing public concern because of their potential adverse health effects. Many pollutants have chemical groups of toxicological importance that can be characterized and detected by Raman spectroscopy. Raman spectroscopy, however, has not been widely used in trace organic detection, even though the information contained in a Raman spectrum is valuable for chemical identification. One limitation of conventional Raman spectroscopy is its low sensitivity, which often necessitates the use of powerful and costly laser sources for sample excitation. Raman spectroscopists have recently been able to analyze dilute biological samples as a result of enhancements in the Raman scattering cross section by factors up to 10(10) when a compound is adsorbed on or near a special electron-conducting surface. These spectacular enhancement factors of the normally weak Raman scattering process help overcome the low sensitivity of Raman spectroscopy through a combination of electromagnetic and chemical interactions between the analyte molecule and the surface. The technique associated with this phenomenon is known as surface enhanced Raman scattering spectroscopy (SERS). The special conductive surface responsible for the scattering enhancement is referred to as a SERS substrate. For the past few years we have developed the SERS technique, using practical SERS active substrate materials based on silver-coated microspheres deposited on glass. A wide variety of biomarkers have been investigated, including benzo[a]pyrene, dibenz[a,h]anthracene epoxides, 1,N9-ethenoadenine, 3,N4 ethenocytosine, and other substances. These biomarkers were measured at nanogram and subnanogram levels. The experimental results are of great analytical interest, since these chemicals are difficult to detect by other techniques, such as luminescence spectroscopy, because of the weak luminescence quantum yields of these DNA adducts. In this paper the potential usefulness of the SERS technique for assessing environmental and health effects from human exposure to toxic pollutants is demonstrated. PMID- 8230296 TI - Biomarkers and molecular epidemiology of occupationally related cancer. AB - Effective prevention of cancer requires sensitive early warning systems to identify groups, and ultimately individuals, who are at high risk of developing cancer and to accurately estimate the magnitude of their risk. Incorporated with molecular epidemiologic studies, biologic markers have the potential to provide quantitative human data on the biologically effective dose of carcinogens, resultant molecular effects, and genetic/acquired factors that modulate these effects. Clearly, this information is directly relevant to risk identification and to risk quantification. PMID- 8230297 TI - Preliminary report on a simple new assay for DNA-protein cross-links as a biomarker of exposures experienced by welders. AB - A method originally developed to detect topoisomerase DNA complexes has been adapted to determine DNA-protein cross-links formed in cells following their exposure in vitro and in vivo to cross-linking agents. A preliminary study on welders and controls has been carried out to assess the feasibility of this assay to detect human exposure to chromium (Cr) and nickel (Ni) that are associated with increased DNA-protein cross-links. The percentage of cross-link was significantly higher among welders (1.85% +/- 1.14) than among controls (1.17% +/ 0.46; p = .01) Cross-links were not affected by age, weight, or smoking status. The assay developed has substantial advantages over previous methods, being considerably more simple, inexpensive, and sensitive. PMID- 8230298 TI - Rationale for selecting exfoliated bladder cell micronuclei as potential biomarkers for arsenic genotoxicity. AB - Biomarkers of effect have important potential in epidemiology, since they may enable ascertainment of exposure-effect associations in relatively inexpensive cross-sectional studies, with confirmation by short follow-up after cessation of exposure. Arsenic is known to cause human skin and lung cancer, and may also cause various internal cancers including bladder, kidney, and liver cancer. The strongest epidemiological association between arsenic ingestion and an internal cancer is that with bladder cancer. Epidemiological studies of a Taiwanese population exposed to high levels of arsenic from drinking water reported relative risks for bladder cancer well above any other known environmental carcinogen. Populations at increased risk for bladder cancer from other exposures, such as smoking and schistosomiasis infection, have elevated frequencies of micronuclei in exfoliated bladder cells. We have therefore proposed that the bladder cell micronucleus assay could be an appropriate biological marker of genotoxic effect of arsenic exposure. In this paper, we present the rationale for choosing the bladder cell micronucleus assay as a potential biomarker of effect for arsenic. We also briefly describe the studies we are conducting using this biomarker in currently exposed populations. PMID- 8230299 TI - Urinary porphyrin profiles as a biomarker of mercury exposure: studies on dentists with occupational exposure to mercury vapor. AB - Porphyrins are formed as intermediates in the biosynthesis of heme. In humans and other mammals, porphyrins with eight, seven, six, five, and four carboxyl groups are excreted in the urine in a well-established pattern. Mercury selectively alters porphyrin metabolism in kidney proximal tubule cells, leading to an altered urinary porphyrin excretion pattern. Previous studies in rats have shown that changes in the urinary porphyrin profile during exposure to mercury as methylmercury hydroxide are uniquely characterized by highly elevated (20- to 30 fold) levels of four- and five-carboxyl porphyrins and by the excretion of an atypical porphyrin ("precoproporphyrin"), which elutes on high performance liquid chromatography (HPLC) approximately midway between penta- and coproporphyrins. Changes in the urinary porphyrin profile are highly correlated with the dose and duration of mercury exposure and persist for up to 20 wk following cessation of mercury treatment. In the present studies, the utility of urinary porphyrin profile changes as a biomarker of mercury exposure in human subjects was evaluated. Urinary porphyrin concentrations were measured in dentists participating in the Health Screening Programs conducted during the 1991 and 1992 annual meetings of the American Dental Association and compared with urinary mercury levels measured in the same subjects. Among dentists with no detectable urinary mercury, mean concentrations of urinary porphyrins were within the established normal ranges for male human subjects. In contrast, among dentists with urinary mercury in excess of 20 micrograms/L, mean urinary concentrations of four- and five-carboxyl porphyrins as well as of precoproporphyrin were elevated three to four times those of unexposed subjects. Significant differences in urinary porphyrin concentrations remained when porphyrin concentrations in spot urine samples were adjusted for creatinine levels. These findings suggest that urinary porphyrin profiles may serve as a useful biomarker of mercury exposure in clinical or epidemiologic studies of mercury-related human health risks. PMID- 8230300 TI - Identification of hydroxyl radical-induced lesions in DNA base structure: biomarkers with a putative link to cancer development. AB - Hydroxyl radical-induced DNA base lesions of guanine and adenine were originally found in neoplastic and microscopically normal livers of fish exposed to environmental carcinogens. They were later identified in a mammalian tissue--the cancerous female breast. This evidence suggested that the base lesions are broadly present in the cancerous and microscopically normal tissues of a variety of eukaryotic organisms. The base lesion concentrations in both neoplastic tissues frequently exceeded 1 modified base in 1000 normal bases. By contrast, the base lesion:normal base ratios in healthy tissues were generally 10-100 times less. A greater variety of base lesions was found subsequently in the cancerous lung, brain, and other human tissues, although information relating to their biological significance is largely confined to the originally found purine derivatives. The biochemistry of the base lesions and the relationship of ring opening (Fapy) derivatives to OH adducts in the DNA of normal and cancerous tissues is discussed with regard to the etiology of cancer and the potential use of the lesions as biomarkers for cancer risk assessment. PMID- 8230301 TI - Brain fibronectin expression in prenatally irradiated mice. AB - Activation of gene transcription by radiation has been recently demonstrated in vitro. However, little is known on the specificity of these alterations on gene transcription. Prenatal irradiation is a known teratogen that affects the developing mammalian central nervous system (CNS). Altered neuronal migration has been suggested as a mechanism for abnormal development of prenatally irradiated brains. Fibronectin (FN), an extracellular glycoprotein, is essential for neural crest cell migration and neural cell growth. In addition, elevated levels of FN have been found in the extracellular matrix of irradiated lung. To test whether brain FN is affected by radiation, either FN level in insoluble matrix fraction or expression of FN mRNA was examined pre- and postnatally after irradiation. Mice (CD1), at 13 d of gestation (DG), served either as controls or were irradiated with gamma rays at 0.5 or 1 Gy. Control and irradiated animals were killed either at 13 DG, 14 DG, 17 DG, or 5, 6, or 14 d postnatal. Brain and liver were collected from offspring and analyzed for either total FN protein levels or relative mRNAs for FN and tubulin. Results of prenatal irradiation on reduction of postnatal brain weight relative to whole body weight and morphological reduction in cerebral cortex regions of postnatal brains are comparable to that reported by others. Insoluble matrix fraction (IMF) per gram of brain, liver, lung, and heart weight was not significantly different either between control and irradiated groups or between postnatal stages, suggesting that radiation did not affect the IMF. However, total amounts of FN in brain IMF at 17 DG were significantly different (p < .02) between normal (1.66 +/- 0.80 micrograms) and irradiated brains (0.58 +/- 0.22 microgram). FN mRNA was detectable at 13, 14, and 17 DG, but was not detectable at 6 and 14 d postnatal, indicating that FN mRNA is developmentally regulated. After 0.5 Gy of irradiation, expression of FN mRNA was reduced to 36% +/- 22% (1 h), 52% +/- 10% (1 d), and 76% +/- 10% (4 d) of the control level. After 1 Gy of irradiation, relative FN mRNA was 62% +/- 28% (1 h) and 75% +/- 3% (4 days) to the control level, respectively. This reduction was comparable to that reported by others for the cytoskeletal protein beta actin. In contrast, mRNA for tubulin, another cytoskeletal protein, increased at 1 h after irradiation but then approached normal postnatally. The longer lasting alteration of FN may be more directly related to neural development, particularly if the reduction in FN is nonuniform. PMID- 8230302 TI - Metaphase chromosome aberrations as markers of radiation exposure and dose. AB - Chromosome aberration frequency provides the most reliable biological marker of dose to detect acute accidental radiation exposure. Significant radiation-induced changes in the frequency of chromosome aberrations can be detected at very low doses (Lloyd et al., 1992). In animal studies chromosome aberrations provide a method to relate exposure to cellular dose. Using an in vivo/in vitro approach, aberrations provided a biological marker of dose from radon progeny exposure, which was used to convert exposure, work level months (WLM) to dose in grays (Gy) delivered to rat tracheal epithelial cells. Injection of Chinese hamsters with 144Ce, which produced a low-dose rate exposure of bone marrow to low-linear energy transfer (LET) radiation, increased the cell sensitivity for the induction of chromatid exchanges by subsequent external 60Co exposure. Our paper provides information on using molecular chromosome probes to "paint" chromosomes and score chromosome damage. This approach illustrates how technical advances make it possible to understand the mechanisms involved in the formation of chromosome aberrations. These studies demonstrate the usefulness of chromosome damage as a biological marker of dose and cellular responsiveness. PMID- 8230303 TI - Role of human cytochrome P-450s in risk assessment and susceptibility to environmentally based disease. AB - Cytochromes P-450 (P-450s) are a large group of heme-containing proteins that carry out oxidation of numerous chemicals. In mammals, a limited number of P-450s are involved in metabolic pathways of steroid synthesis, while most of these enzymes are involved in metabolism of foreign compounds. The principal beneficial function of P-450s is to convert chemicals into derivatives that can be easily eliminated from the body. This generally occurs through P-450-mediated oxidations of hydrophobic substances followed by conjugation reactions. For many foreign compounds, P-450 metabolism results in production of "activated" metabolites that can cause cell death and gene mutations. During the past several years, it has become widely recognized that marked species differences occur among the foreign compound-metabolizing P-450s. In addition to this interspecies variability in metabolism, marked intraspecies variability, frequently referred to as drug oxidation polymorphisms, occurs in virtually all mammals examined to date. Based on these observations, it is necessary to develop new human P-450-based systems that can be used to study foreign compound metabolism in order to predict human risk. This is being accomplished by use of cDNA-directed expression in B lymphoblastoid cells. These cells can be used to predict how humans will metabolize a chemical and whether it will be metabolically activated to a toxic or mutagenic metabolite. To study human P-450 polymorphisms, polymerase chain reaction (PCR) assays have been developed for diagnosis of known mutant P-450 genes. Molecular probes are also being used to screen populations for levels of expression of carcinogen-activating P-450s in an effort to determine whether expression of certain P-450 forms is associated with increased risk for development of environmentally based disease. PMID- 8230304 TI - Molecular markers and the epidemiology of insulin-dependent diabetes mellitus worldwide. PMID- 8230305 TI - Novel biomarkers of genetic damage in humans: use of fluorescence in situ hybridization to detect aneuploidy and micronuclei in exfoliated cells. AB - Here we describe new techniques that employ fluorescence in situ hybridization (FISH) with centromeric and chromosome-specific DNA probes to detect aneuploidy and micronucleus formation in exfoliated human epithelial cells. Micronuclei arise from chromosome breakage or lagging, which results in chromosome fragments and whole chromosomes being left outside of the main nucleus at telophase. By using a centromeric DNA probe to detect the presence of whole chromosomes in micronuclei and propidium iodide as a general DNA stain in exfoliated nasal, buccal, and bladder cells, we have developed a new fluorescent method that can detect micronuclei and determine the mechanism of formation. The new fluorescent technique gave results that were very similar to those obtained with the standard Feulgen-fast green method. The spontaneous levels of micronuclei in healthy volunteers were buccal, 0.13%, nasal, 0.21%, and urothelial, 0.07%, in approximately 1500 cells per data point. These values are lower than that found in cultured lymphocytes, 0.4-0.8%. Approximately 50% of the exfoliated cell micronuclei contained whole chromosomes (centromeric DNA). FISH was also used to detect aneuploidy in exfoliated buccal and bladder cells. A DNA probe specific for chromosome 9 was used. Average frequencies for 0, 1, 2, 3, and 4 hybridization regions were 4.8, 9.3, 84.8, 0.8, and 0.3% for urothelial cells and 8.2, 9.9, 80.1, 1.4, and 0.4% for buccal cells. The estimated frequency of aneuploidy in exfoliated cells is similar to that found in human lymphocytes analyzed by FISH with the same probe for chromosome 9. These techniques are potentially useful for epidemiological studies of exposed populations and are currently being applied in our laboratory for studies of arsenic- and formaldehyde-exposed populations. PMID- 8230306 TI - Use of biological markers in occupational health research and practice. AB - The promise of biological markers in occupational health research and practice has been described in the scientific literature. The current generation of biological markers has the potential to allow for the earlier detection of disease, for the reduction of misclassification of exposure and outcome, for heightened understanding of mechanisms and etiologic pathways, and for the designation of groups at risk. What is necessary now is a strategy for realizing this potential. The elements of such as a strategy may include the following: (1) a program to validate biomarkers, (2) increased utilization of valid biomarkers in etiologic and prevention research, and (3) developmental programs to encourage interdisciplinary collaboration and train molecular epidemiologists. A framework for linking biomarkers and epidemiologic study designs has evolved during the past 5 yr. For this progress to continue, it is important that discussion about biomarkers reflect a specificity with regard to both the type of marker and the use for which it is intended. PMID- 8230307 TI - Effect of in vivo exposure to iodine-131 on the frequency and persistence of micronuclei in human lymphocytes. AB - The validity of the micronucleus test as a biomarker of chromosome damage in dividing mammalian cells is well established. This assay was used to study the response of peripheral lymphocytes of a 34-yr-old male patient following treatment with 131I ablative radiation therapy following a total thyroidectomy. Coincidentally, 8 mo before diagnosis, the patient had provided a blood sample for an in vitro study of micronucleus induction following exposure to graded doses of x-rays. The background frequency in the unexposed culture showed a mean count of 6.0 micronuclei per 1000 binucleated (first division) lymphocytes, while mean values of 18.5, 29.0, 41.0, 61.0 and 75.5 micronuclei/1000 cells were observed following x-ray doses of 5, 10, 15, 20, and 25 cGy, respectively. These data fit a nonthreshold, linear dose-response function (y = 2.78x + 3.71; r = .99). Eight months after the in vitro x-ray study, the subject was diagnosed with thyroid cancer. Surgery was performed, and 5 wk later the patient received 1.78 GBq (48 mCi) of 131I as adjuvant radiation therapy. Blood was drawn 11 d after the radiation treatment and at monthly intervals thereafter to analyze the frequency and persistence of micronuclei. The first posttreatment sample showed 35.5 micronuclei per 1000 binucleate cells. Based on the linear dose-response equation from the earlier study, the sixfold increase in micronucleus frequency suggests a dose to the peripheral blood of approximately 11 cGy. The cytogenetic dose estimate compares to approximately 30 cGy using a new model based on external whole-body counting data. Nine consecutive monthly samples have been analyzed to date. Although the micronucleus count has fluctuated (four- to sixfold above background), the frequency after 8 mo is equivalent to the first posttreatment sample. Data show that radiation-induced cellular lesions persist for months following relatively brief radiation exposure to a medical isotope. Results of this study support the conclusion that the lymphocyte micronucleus test is a rapid, sensitive, and perhaps quantitative biomarker of low-dose (< 25 cGy) radiation exposure. PMID- 8230308 TI - Biomarkers of human exposure to benzene. AB - Three biomarkers for benzene exposure were developed. The first biomarker, muconic acid in urine, results from the ring opening of a benzene metabolite. A gas chromatography/mass spectroscopy (GC/MS) assay was developed to measure urinary muconic acid, and the analyte in urine samples from workers occupationally exposed to benzene was determined. Workers exposed to benzene concentrations as low as 4.4 ppm over an 8-h day showed higher urinary muconic acid concentrations than did any control individual (p < .005). The second biomarker, S-phenylcysteine (SPC) in hemoglobin (Hb), results from the addition of benzene oxide to a cysteine sulfhydryl group. A GC/MS assay was developed to measure SPC in the blood of F344/N rats and B6C3F1 mice exposed to benzene by inhalation. The cysteine moiety on rat Hb is at a more accessible site than on Hb of mice or humans, and rats showed considerably higher levels of SPC than did mice. As yet, we have been unable to detect SPC in the globin of humans occupationally exposed to benzene. The third biomarker is SPC in albumin. In humans occupationally exposed to average concentrations of 0, 4.4, 8.4, and 23.1 ppm benzene, 8 h/d, 5 d/wk, SPC increased in the exposed groups linearly, giving a statistically significant slope (p < .001) of 0.044 +/- 0.008 pmol/mg albumin/ppm. The assay for SPC is arduous and often imprecise; assuming these difficulties can be overcome, muconic acid in urine and SPC in albumin may be useful for accurately determining benzene exposure. PMID- 8230309 TI - Variables affecting performance on human behavioral neurotoxicology test batteries: education, culture, gender, age, presentation mode. PMID- 8230310 TI - 8-Hydroxydeoxyguanosine as a urinary biomarker of oxidative DNA damage. AB - Living organisms are continuously exposed to reactive oxygen species as a consequence of biochemical reactions as well as external factors. Oxidative DNA damage has been implicated in aging, carcinogenesis and other degenerative diseases. The urinary excretion of the DNA repair product 8-hydroxydeoxyguanosine (8OHdG) has been proposed as a noninvasive biomarker of oxidative DNA damage in humans in vivo. We have developed a three-dimensional HPLC analysis with electrochemical detection for the analysis of 8OHdG in urine and studied factors affecting the excretion of this biomarker in 83 healthy humans and in various laboratory animals, including dog, pig, and rat. Previously, other groups have used comparable HPLC methods or gas chromatography-mass spectrometry with selective ion monitoring for measuring the excretion of 8OHdG in humans, rats, mice, and monkeys. In the 169 humans studied so far, the average 8OHdG excretion was 200-300 pmol/kg per 24 h with a sevenfold range, and the coefficient of variation was 30-40%. This excretion corresponds 140-200 oxidative modification of guanine bases per cell per day. Thirty-two smokers from our study population excreted 50% (31-69%; 95% confidence interval) more 8OHdG than 53 nonsmokers. This indicates a 50% increased rate of oxidative DNA damage from smoking, adding to the other well-known health hazards of smoking. The biochemical-physiological basis is unknown but may be related to smoke constituents including or generating reactive oxygen species and/or consuming antioxidants and/or the well-known enhancing effect of smoking on the metabolic rate. In our 83 healthy subjects the 8OHdG excretion correlated with body composition. Thus, lean and/or male subjects excreted more than obese and/or female subjects, possibly related to differences in metabolic rate. In accordance, the excretion of 8OHdG decreased after calorie restriction, which will cause a decline in the metabolic rate. Across the investigated species, humans, dogs, pigs, and rats, the excretion of 8OHdG correlated with the specific metabolic rate, confirming data from other groups on humans, monkeys, rats, and mice. The excretion of 8OHdG decreased with age in rats in parallel with the decline in metabolic rate with advancing age. The excretion of 8OHdG reflects the formation and repair of only one out of approximately 20 described oxidative DNA modifications. So far, methods are not available for the determination of the corresponding repair products, except 8OHdG and thymidine glycol, in urine. Moreover, the importance in terms of mutagenicity, particularly regarding tumour suppressor genes and oncogenes, is mainly documented for 8OHdG in DNA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8230311 TI - Frequency of micronuclei in lymphocytes from a group of floriculturists exposed to pesticides. AB - A large part of the commercial flower production in Italy is located in the located in the western part of the Liguria region, near the French border. The use of pesticides in this area has been much higher than the national average. The frequency of micronuclei in peripheral blood lymphocytes was evaluated in a group of 71 floriculturists working in this area and in a control group of 75 healthy blood donors living in the same area. A significant increase of micronucleated lymphocytes was observed in floriculturists as compared to unexposed subjects (8.57 vs. 6.67, p < .05). Females showed a marked increase in MN frequency (45% higher than males) independently of the exposure. A dose response relationship was observed between duration of exposure and MN frequency. The condition of exposure was also found to influence the micronuclei frequency. Increased relative risks in greenhouse workers (RR = 1.31) and in people working alternately in the greenhouse and in the open field (RR = 1.46) was observed with respect to the reference population. PMID- 8230312 TI - Two-stage evaluation of exposure to mercury and biomarkers of neurotoxicity at a hazardous waste site. AB - Communities surrounding the Rocky Mountain Arsenal (RMA), a Superfund site in Colorado, were studied in order to determine whether exposures to mercury were greater among persons who resided there than among residents of a comparison area 12-15 miles distant. From a census-based stratified random sample, 469 persons were interviewed and urine samples were obtained for biomonitoring. Mercury was detected in urine from 32 (6.8%) of the 469 persons sample at a detection limit of 5 ppb. Trace levels of mercury (detectable, but nonquantifiable) were found in 80 (17.1%) of the persons sampled. Neither the frequency of detection, the arithmetic mean, nor the geometric mean value for urine mercury was found to be statistically different when persons living near the site were compared to persons from the more distant comparison area. The risk of mercury exposure associated with demographic variables, residence, occupation, hobbies, dietary habits, water supply, housing, and activity patterns was evaluated. In the second stage of the evaluation, the Neurobehavioral Core Test Battery (NCTB) is being used to assess individual functional deficits and nervous system disorders associated with exposure to mercury and other neurotoxic chemicals. PMID- 8230313 TI - Classification of adverse reproductive effects can be improved by measurements of multiple biomarkers for ovarian toxicity and early fetal loss. AB - Hormone assays have been developed and applied for monitoring reproductive function using self-collected urine samples in non-clinical populations of women. Early pregnancy loss, menstrual dysfunction, reduced fertility as well as the site of toxicity can now be assessed using daily early morning urine samples. The understanding of the specific limitations of individual assay systems is important, however, to make the best use of these systems. The use of multiple end-points and computer algorithms is suggested to avoid misclassification of adverse reproductive events. PMID- 8230314 TI - Interaction between epidemiology and laboratory sciences in the study of birth defects: design of birth defects risk factor surveillance in metropolitan Atlanta. AB - Despite years of research, the etiology of most birth defects remains largely unknown. Interview instruments have been the major tools in the search for environmental causes of birth defects. Because of respondents' problems with recognition and recall, interviews are limited in their capacity to measure certain exposures. Laboratory scientists can have a major impact on defining markers of environmental exposure and genetic susceptibility. The Centers for Disease Control is starting a case-control study of serious birth defects on the basis of a population-based surveillance system for birth defects diagnosed during the first year of life in metropolitan Atlanta. Each year, 300 infants with selected birth defects (case subjects) and 100 population-based control subjects (infants without birth defects) will be enrolled in an ongoing study that will supplement surveillance. In addition to conducting extensive maternal interviews, we will collect blood and urine specimens from case and control subjects and their mothers for laboratory testing. Eventually, some environmental sampling may be incorporated. Particular areas of emphasis are (1) nutritional factors, specifically measuring maternal folic acid levels and other micronutrients (e.g., zinc) to explore their role in the etiology of neural tube defects, (2) substance use, specifically measuring cocaine metabolites in the blood and urine to explore their role for specific vascular disruption defects, and (3) environmental factors such as pesticides and aflatoxins, to explore their potential relationships with specific defects. In addition, a DNA bank will be maintained to evaluate the role of specific candidate genes in the etiology of birth defects. The development and testing of these methods could be useful to assess the interaction between environmental exposures and genetic susceptibility in the etiology of birth defects. PMID- 8230315 TI - Herbert M. Parker Lecture. The risk of radiation induced cancer--whence? Whither? PMID- 8230316 TI - Nerve growth factor and p75NGFR factor receptor mRNA change in rodent CNS following stress activation of the hypothalamo-pituitary-adrenocortical axis. AB - The synthesis of nerve growth factor (NGF) by the hippocampus raises the possibility that NGF may play a role in the regulation of the hypothalamic pituitary-adrenal axis (HPAA). Subchronic cold stress has been shown to activate the HPAA in a mild noninvasive manner, to stimulate serum glucocorticoid levels, and to perturb NGF binding in hippocampus and basal forebrain. One or repeated episodes of cold stress increased NGF mRNA levels in the hippocampus and p75NGFR mRNA levels in the basal forebrain. These changes were not due to elevated serum glucocorticoid levels since treatment with exogenous corticosterone had no effect on NGF and p75NGFR mRNA levels. Adrenalectomy did not prevent the stress induced increases in NGF and p75NGFR mRNA. PMID- 8230317 TI - Expression of three nonallelic genes coding calmodulin exhibits similar localization on the central nervous system of adult rats. AB - By Northern blot analysis with the digoxigenin-labeled antisense RNA probes of the noncoding regions, the transcripts of three calmodulin (CaM) genes, CaMI, CaMII, and CaMIII, were separately detected in 12 different tissues of adult Wistar albino rats, without any cross-hybridization. The mRNAs of all three CaM genes were abundant in the central nervous system (CNS) as well as in the testis, although ubiquitous expression was detected at low levels in the other tissues. There were subtle but significant differences in the tissue-specific distribution of the three CaM gene RNAs. By in situ hybridization, strong hybridization of the three CaM gene probes was observed in common in large projection neurons of the CNS: the hippocampal pyramidal cells, the cerebellar Purkinje cells, and the large neurons of the cerebral neocortex, the pyriform cortex, the mesencephalon, the pons, and the spinal cord. The expression of the three CaM genes was at lower levels in small interneurons of the CNS. These profiles of expression were almost the same among the three CaM genes. Thus, all three CaM genes were coordinately expressed in neurons of the adult rat CNS. Certain regulatory mechanisms of the three CaM genes seemed to mediate similar tissue- and cell type-specific expression in the CNS. PMID- 8230318 TI - Expression of the metabotropic glutamate receptor mGluR1 alpha and the ionotropic glutamate receptor GluR1 in the brain during the postnatal development of normal mouse and in the cerebellum from mutant mice. AB - Expression of the metabotropic glutamate receptor type 1 alpha (mGluR1 alpha) and the non-N-methyl-D-aspartate (NMDA) ionotropic glutamate receptor type 1 (GluR1) in mouse brain was investigated using the antibodies raised against the synthetic peptides corresponding to their C-terminal amino acid sequences. Both receptor proteins are glycosylated predominantly in an asparagine-linked manner, and are abundant in post-synaptic membranes. We showed that mGluR1 alpha and GluR1 expression within the first 3 postnatal weeks undergoes dramatic changes in time and space, i.e., in the hippocampus and cerebellum. These spatio-temporal expression patterns appear to be correlated with the postnatal ontogenesis and establishment of the glutamatergic neurotransmission system in the hippocampus and cerebellum, cell migration, dendritic and axonal growth, spine formation, and synaptogenesis. In the adult cerebellum, mGluR1 alpha is intensely expressed in Purkinje neurons and GluR1 in Bergmann glial cells. Both receptors are expressed to a fair degree in weaver mutant cerebellum despite granule cell degeneration. However, the intrinsic expression levels of both mGluR1 alpha and GluR1 are markedly reduced in the cerebellum of the Purkinje cell-deficient and underdeveloped mutant mice, Purkinje-cell-degeneration, Lurcher, and staggerer, suggesting that GluR1 expression in Bergmann glia cells may be correlated with the sustained interaction with adjacent Purkinje neurons. PMID- 8230319 TI - Purification and characterization of NCAD90, a soluble endogenous form of N cadherin, which is generated by proteolysis during retinal development and retains adhesive and neurite-promoting function. AB - The cadherins are calcium-dependent cell adhesion molecules which regulate cell cell interactions during morphogenesis. During development, cadherin expression is subject to dynamic patterns of regulation. We have previously demonstrated that expression of N-cadherin, the predominant cadherin of neural tissues, is sharply down-regulated during development of the retina and brain during later stages of histogenesis (Lagunowich and Grunwald, Dev Biol 135:158-171, 1989; Lagunowich et al., J Neurosci Res 32:202-208, 1992), and that this down regulation is due to multiple factors, including decreased mRNA levels and turnover apparently mediated by endogenous metalloproteolytic activity (Roark et al., Development 114:973-984, 1992). In the present study, we describe metabolic studies which provide direct biochemical evidence for turnover of 130-kDa N cadherin in embryonic retina tissues, yielding a soluble 90-kDa N-terminal fragment. We demonstrate that this form of N-cadherin, which we refer to as NCAD90, accumulates in vivo during development. We further demonstrate that purified NCAD90, obtained from embryonic vitreous humor, retains biological function and promotes cell adhesion and neurite growth in a dose-dependent fashion among chick embryo neural retina cells when present in a substrate-bound form. The morphology of retinal cells and neurites grown on a substrate of NCAD90 differs strikingly from that seen on a laminin substrate, in a manner similar to that described for intact 130-kDa N-cadherin. We conclude that proteolysis of N cadherin at the cell surface during embryonic retinal histogenesis is an endogenous mechanism for regulating N-cadherin expression which generates a novel and functional form of the protein. The results further indicate that an intact cytoplasmic domain is not essential for all cadherin functions. PMID- 8230320 TI - Protein growth factor requirements of rat neural crest cells. AB - Nascent neural crest cells derived from explanted E12 embryonic rat caudal neural tubes were used as an assay system to investigate the effects of fibroblast growth factors on neural crest cell (NCC) survival, proliferation, migration, and differentiation. In vitro and in vivo all NCC express low affinity nerve growth factor receptors (p75-LNGFR), whereas a subpopulation of NCC expresses the carbohydrate epitope recognized by the monoclonal antibody HNK-1 (Bannerman and Pleasure, manuscript in preparation). Both acidic and basic fibroblast growth factor (FGF) promoted the survival of proportionally greater numbers of p75 LNGF+/HNK-1- than P75-LNGFR+/HNK-1+ NCC. An as yet uncharacterized factor present in neural tube-conditioned medium was also required for NCC survival. Mitosis was frequent in those NCC closest to the neural tube, less so as the cells migrated away. Neither basic nor acidic fibroblast growth factor (FGF) influenced rates of NCC mitosis in either of these locations, nor did these FGFs alter the rate at which nascent NCC migrated away from the neural tube. However, acidic and basic FGFs did delay the differentiation of neural crest derived neurons in the cultures. FGF is abundant in the embryonic rat neural crest outgrowth zone, and the present study strongly supports an essential role for FGF in early development of the mammalian neural crest. PMID- 8230321 TI - Early responses of PC-12 cells to NGF and EGF: effect of K252a and 5' methylthioadenosine on gene expression and membrane protein methylation. AB - Although epidermal growth factor (EGF) and nerve growth factor (NGF) have markedly different biological effects on PC-12 cells, many of the signaling events following ligand binding are similar. Both EGF and NGF result in the induction of the primary response gene egr-1/TIS8 and increased methylation of a variety of membrane-associated proteins as early as 5 min after EGF or NGF treatment using a methylation assay that detects methyl esters as well as methylated arginine residues. At 20 min after stimulation with these factors, the stimulation of methylation by NGF is greater than that of EGF, especially in the polypeptides of 36-42 and 20-22 kDa. To help dissect the pathways involved in these cellular responses, the protein kinase inhibitor K252a and the methyltransferase inhibitor 5'-methylthioadenosine (MTA) were used. Both K252a and MTA inhibit NGF-, but not EGF-mediated, primary response gene expression. In contrast, MTA, but not K252a, can block NGF-induced membrane associated protein methylation. These data suggest a role for differential protein methylation reactions in EGF and NGF signal transduction. PMID- 8230322 TI - Transient requirement for vimentin in neuritogenesis: intracellular delivery of anti-vimentin antibodies and antisense oligonucleotides inhibit neurite initiation but not elongation of existing neurites in neuroblastoma. AB - Vimentin is initially expressed by nearly all neuronal precursors in vivo, and is gradually replaced by neurofilaments shortly after the immature neurons become postmitotic (Cochard and Paulin, 1984, J Neurosci 4:2080; Tapscott et al., 1981, Dev Biol 86:40). A transient increase in neuritic vimentin filaments occurs within the first day of dbcAMP-mediated neurite induction in NB2a/d1 neuroblastoma, after which vimentin levels rapidly decline and neurofilaments increase (Shea, 1990, Brain Res 521:343). In the present study, we tested the possibility that vimentin filaments may function in neurite elaboration by inducing neuritogenesis under conditions where vimentin expression and assembly was inhibited. Intracellular delivery of anti-vimentin antiserum into transiently permeabilized NB2a/d1 cells prevented the initial elaboration of neurites, but did not retract existing neurites. By contrast, intracellular delivery of antiserum directed against the low molecular weight neurofilament subunit or normal rabbit antiserum did not affect neurite outgrowth. Treatment with vimentin antisense oligonucleotides reversibly depleted vimentin synthesis and steady state levels, and prevented neurite initiation, but did not induce retraction of existing neurites. These findings point toward an hitherto undetected role for vimentin in the initiation of neurite outgrowth. PMID- 8230323 TI - Protein kinases modulate the sensitivity of hippocampal neurons to nitric oxide toxicity and anoxia. AB - Multiple processes lead to neuronal death after ischemia, but the generation of nitric oxide (NO) is a key component in this cascade of events. The mechanisms that regulate the extent of neuronal degeneration during anoxia and NO toxicity are multifactorial. Neuronal death may be modulated by the activity of signal transduction systems that influence the toxicity of NO or its metabolic products such as cGMP. The enzyme responsible for the production of NO, nitric oxide synthase (NOS), is phosphorylated by protein kinase C (PKC), the cAMP-dependent protein kinase (PKA), and the calcium/calmodulin-dependent protein kinase II (CaM II). We examined in primary cultured hippocampal neurons whether the protein kinases PKC, PKA, CaM-II, and cGMP-dependent protein kinase modified the toxic effects of anoxia and NO. Down-regulation of PKC activity with PMA (1 microM) increased hippocampal neuronal survival during anoxia and NO exposure from approximately 22% to 88%. Inhibitors of PKC activity (H-7, H-8, sphingosine, and staurosporine) also were neuroprotective. Down-regulation of PKC activity increased survival during anoxia even in the presence of the NOS inhibitor, N omega-methyl-L-arginine. Thus, although down-regulation of PKC activity may increase neuronal survival by decreasing NOS activity, it also is likely that PKC contributes to ischemic neuronal death by mechanisms that are independent of NOS. Inhibition of the cGMP-dependent protein kinase activity, but not the activity of the CaM-II also was neuroprotective during NO administration. In contrast to the protective effects of inhibition of PKC and the cGMP-dependent protein kinase, activation rather than inhibition of PKA increased hippocampal neuronal survival during NO exposure. These results indicate that neuronal survival during anoxia and NO exposure is linked to the modulation of PKC, PKA, and cGMP-dependent protein kinase activity but is not dependent on the CaM-II pathway. Understanding the involvement of PKC, PKA, and the cGMP-dependent protein kinase in modulating the effect of neuronal death during ischemia and NO toxicity may help in directing future therapeutic modalities for cerebrovascular disease. PMID- 8230324 TI - Intracellular distribution of transgenic bacterial beta-galactosidase in central nervous system neurons and neuroglia. AB - Bacterial beta-galactosidase is widely used as a marker for gene expression and in cell tracing experiments. In a survey of three transgenic mouse lines expressing beta-galactosidase in the central nervous system (CNS) under the control of different promoters, we find substantial variation in the intracellular distribution of the lacZ protein. In line M beta P5, transgene beta galactosidase expression is driven by a promoter/enhancer fragment from the oligodendrocyte-specific myelin basic protein gene; however, electron microscopy of histochemically stained preparations reveals transgene expression not only in oligodendrocytes but also in some neurons. Immunofluorescence and immunoperoxidase staining show the beta-galactosidase protein distributed throughout the perikaryal cytoplasm of oligodendrocytes and in processes reaching to myelin sheaths. By contrast, immunoreactive protein appears restricted in neurons to one or a few small perikaryal immunoreactive granules. The granules are visible in the electron microscope as amorphous inclusion bodies of moderate electron density and lack a limiting membrane. Histochemical staining patterns with X-gal and Bluo-gal echoed the protein distribution: diffuse distribution of enzyme protein yielded cells filled with substrate, while punctate enzyme distribution yielded restricted or punctate histochemical staining. Examination of two other lines using different promoter/enhancers to drive expression in the CNS showed both diffuse and punctate beta-galactosidase immunolocalization and histochemical staining. The amount of protein synthesized or other properties, yet unidentified, intrinsic to the target cells may determine the intracellular distribution of beta-galactosidase. In retroviral marking studies, clone members have been identified as those cells filled with X-gal reaction product. This approach may underestimate both clone size and the minimum number of divisions separating the members of each clone. PMID- 8230325 TI - Violence: America's uncivil war--presidential address, Sixth Scientific Assembly of the Eastern Association for the Surgery of Trauma. PMID- 8230326 TI - Experimental volume replacement through lower extremity veins. AB - Hypovolemic shock was produced in anesthetized pigs by removal of 40% of blood volume over 10 minutes. Following blood loss, the inferior vena cava (IVC) was occluded below the renal veins to simulate the hemodynamics of emergency surgical treatment. Control animals were not treated. Experimental animals received intravenous lactated Ringer's solution equal to three times the blood loss given through catheters either in the IVC or the superior vena cava (SVC) to determine if lower extremity access would be efficacious in this model. To define the path taken by the resuscitation fluids, an additional group of animals received technetium-99m-labelled crystalloid through lower extremity catheters with continuous recording of isotope counts in the IVC and right atrium. The treated animals in all experimental groups had significant improvements in mean arterial pressure, cardiac output, and pH compared with controls. There was no significant difference in hemodynamic response in animals receiving volume replacement through the IVC compared with the SVC. When fluid was infused below a clamped IVC, the arrival of isotope in the right atrium was delayed only 1.5 seconds. We conclude that in a model simulating emergency control of potentially lethal hemorrhage, the beneficial effects of fluid resuscitation are unrelated to the site of venous access. Lower extremity veins provide a valuable site for volume replacement even with IVC occlusion. These findings should have direct application to resuscitation and surgical care of seriously injured patients. PMID- 8230327 TI - Acute mortality associated with injuries to the pelvic ring: the role of early patient mobilization and external fixation. AB - PURPOSE: To analyze the effect on mortality of a protocol for early mobilization with external fixation of patients with pelvic ring injuries. METHODS: From 1981 through 1988, 605 patients with pelvic ring fractures and dislocations were treated. In 1982, a protocol for early external fixation of hemodynamically unstable patients and those with structurally unstable pelvic fracture patterns to achieve early mobilization to an upright chest position was initiated. Mortality rates were compared between 1981 (pre-protocol), 1982 (transitional), and 1983 through 1988, after initiation of a protocol of care that included external fixation of the pelvic injury. No statistical changes occurred from 1983 through 1988. RESULTS: Mortality rates in pelvic ring injury patients fell from 26% in 1981, to 6% in 1983 through 1988 (p < 0.001), whereas during the study period the mean injury Severity Score (ISS), 23, did not change. The mortality rate of a group of consecutive patients with comparable ISSs, but without pelvic ring injuries did not change. The mortality rate in patients with systolic blood pressure < 100 mm Hg at admission fell from 41% in 1981 to 21% 1983 through 1988 (p = 0.0001). Mortality in patients with closed head injuries associated with pelvic ring injuries fell from 43% in 1981 to 7% from 1983 through 1988 (p = 0.0001). The proportion of patients undergoing external fixation rose from 3% in 1981 to 31% in 1983 through 1988 (p = 0.0001). CONCLUSIONS: An organized protocol including external fixation and early patient mobilization to an upright chest position reduced mortality associated with injuries of the pelvic ring. Orthopedic stabilization of major skeletal injuries should be viewed as part of patient resuscitation, not reconstruction. PMID- 8230328 TI - Immediate internal fixation of low-velocity gunshot-related femoral fractures. AB - Fractures caused by gunshots are increasingly common in urban hospitals and trauma centers. The rising incidence and complexity of these injuries present difficult management problems and health care burdens. In a 3-year period, from 1989 through 1991, 21 patients with femoral shaft fractures from low-velocity bullets were treated with intramedullary fixation within 15 hours of admission. Eighteen patients were available for follow-up. The fractures had healed in all patients. Average hospitalization for an isolated injury was 7 days. There were no complications related to immediate internal fixation. Immediate internal fixation of femoral shaft fractures caused by low-velocity gunshots can be performed in an efficient and cost effective manner. PMID- 8230329 TI - Very hot intravenous fluid in the treatment of hypothermia. AB - The efficacy and safety of very hot (65 degrees C/149 degrees F) intravenous fluid (IVF) were compared with those of conventional warm (38 degrees C/100.4 degrees F) IVF in the treatment of hypothermia. Eight anesthetized beagles (11-20 kg) were studied. Blood pressure (BP), pulse (P), and core temperature (cT degrees) were recorded at baseline, during hypothermia, and during rewarming. The plasma free hemoglobin (PFHg) was measured to assess hemolysis. Each subject was cooled to 32 degrees C/89.6 degrees F and assigned to receive either 65 degrees C or 38 degrees C IVF via a specially designed multiport balloon-tipped catheter in the superior vena cava (SVC). The IV fluid rate was 80% of the blood volume per hour. Conventional rewarming methods were used in all subjects. After 2 hours the subjects were killed and the SVC was examined for injury. The rate of rewarming was 2.9 degrees C/hour in the 65 degrees C IVF group and 1.25 degrees C/hour in the 38 degrees C IVF group. The cT degrees was significantly different in all subjects after 1 (35.2 degrees +/- 1.03 degrees C vs. 33.2 degrees +/- 0.5 degrees C; p < 0.006) and 2 (37.6 degrees +/- 1.17 degrees C vs. 34.3 degrees +/- 0.9 degrees C; p < 0.004) hours of rewarming. The BP, P, and PFHg were not different. Visual examination of the SVC revealed two lesions in the 65 degrees C IVF group and one in the 38 degrees C group. Mechanical or thermal injury could not be differentiated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230330 TI - In vitro comparison of heated saline-blood admixture with a heat exchanger for rapid warming of red blood cells. AB - We compared the rapidity of rewarming and infusing red blood cells (RBCs) mixed with 24 degrees C, 50 degrees C, and 60 degrees C saline with the rapidity of administering undiluted RBCs through a heat exchanger. We measured flow rate, final temperature, and hemolysis in matched 41-45-day-old pooled AS-1 RBCs infused through high-flow tubing via a 14-gauge catheter under the influence of gravity. Undiluted RBCs were tested as controls. The final temperature of the 60 degrees C admixture technique was lower than that with the heat exchanger (28.5 +/- 0.2 degrees C vs. 32.7 +/- 0.2 degrees C), but the flow rate was higher (258 +/- 8 mL/min vs. 61 +/- 4 mL/min). Admixture with 60 degrees C saline resulted in no increase in hemolysis. This technique appears to be a simple, inexpensive method for rapid rewarming and infusion of RBCs and may be valuable for administration of RBCs simultaneously through multiple sites during resuscitation. PMID- 8230331 TI - Reassessing the role of arteriograms in the management of posterior knee dislocations. AB - Vascular injury has been reported in up to one third of patients with posterior knee dislocations, which has led to the routine use of arteriograms in the management of these injuries. Recent studies have shown physical examination (PE) is reliable in detecting significant vascular injuries requiring surgery from other mechanisms. We hypothesized that PE would be similarly sufficient to assess popliteal injury in patients with posterior knee dislocations. To test this, we reviewed the records of all 37 patients with 38 such injuries at our institution over the past 5 years. The average age was 29.5 years, and 31 patients (81.5%) had other associated injuries. All dislocations were the result of blunt trauma. Two patients (5.3%) had hard signs (distal ischemia and no pulses) that clearly indicated vascular injury. Total occlusions of the popliteal artery were seen on arteriograms in both cases and these were successfully treated surgically with reversed saphenous vein bypass grafts. The remaining 36 dislocations manifested no hard signs of vascular injury (absent pulses, distal ischemia, active bleeding, bruit/thrill). Nineteen patients (50.0%) had normal vascular examination results, did not receive arteriograms, and had no adverse sequelae, with a mean follow-up of 9.3 months (range 1 day-43 months). Sixteen patients with 17 dislocations (44.7%) underwent arteriography and the findings appeared normal in ten extremities; nine of these extremities had normal pulses and one had a diminished but palpable pulse. A minimal injury (intimal defect, 3; narrowing, 4) was demonstrated in seven extremities, five with normal pulses and two with diminished pulses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230332 TI - Serum tumor necrosis factor-alpha profile in trauma patients. AB - Tumor necrosis factor-alpha (TNF-alpha) has been implicated in several late consequences of trauma such as sepsis, multiple organ failure, and ischemia reperfusion injury. However, no data are available to indicate whether TNF-alpha is involved in the initial pathophysiologic response to trauma. To address this issue, serum TNF-alpha was determined (by ELISA) longitudinally (first blood sample on admission) in 100 randomly selected trauma patients admitted to the emergency department and trauma division at Jefferson Medical Center, Philadelphia. The TNF-alpha levels were detectable at one or more time points in 35 patients. Mean values tended to be elevated (50.3 +/- 11.5 pg/mL) during the first 5 days, but this trend did not differ statistically from that in healthy controls (n = 12) and did not correlate with the severity of injury (Injury Severity Score and Glasgow Coma Scale score). The TNF-alpha response was not dependent on the mechanism and site of injury, the presence of shock (systolic blood pressure < 90 mm Hg), and the need for emergent surgery. Also, serum TNF alpha levels were not significantly elevated in patients who subsequently developed multiple organ failure (n = 4), septic shock (n = 5), or both (n = 3). Taken together, these data do not support a role for circulating TNF-alpha in the initial acute inflammatory response to trauma. PMID- 8230333 TI - Diltiazem preserves hepatic gluconeogenesis following hemorrhagic shock. AB - Prolonged hemorrhagic shock is characterized by the progression from hyperglycemia to hypoglycemia and failure to respond to standard methods of resuscitation. Previous studies have shown that the transition to irreversible shock is accompanied by attenuation of hepatic gluconeogenic capacity and a rising level of intracellular calcium. Additionally, it has been observed that diltiazem improves survival following prolonged hemorrhagic shock in rats. We examined the effect of resuscitation containing diltiazem upon hepatic gluconeogenesis during early and late phases of hemorrhagic shock in a rat model. Fasted male Sprague-Dawley rats (250-350 g) were rapidly bled to a mean arterial pressure of 40 mm Hg for a period of 30 minutes (group A) or 120 minutes (group B). At the end of the hemorrhagic shock period, rats were randomized to resuscitation utilizing lactated Ringer's (LR) solution, or LR+diltiazem (DZ, 1.2 mg/kg). Following resuscitation, rats underwent laparotomy and in situ liver perfusion with an oxygenated 37 degrees C glucose-free Krebs solution via the portal vein. After equilibration, 5 mmol/L lactate and 0.5 mmol/L pyruvate were added to the perfusate as substrate and effluent samples collected. Serum glucose concentration and portal venous flow did not differ significantly between DZ and LR groups throughout the study periods. In group A, hepatic glucose production was significantly elevated in DZ animals when compared with controls (p < 0.05). A similar significant improvement in gluconeogenesis was observed following 120 minutes of hemorrhagic shock in group B (p < 0.05). Additionally, treated rats (DZ, both groups A and B) demonstrated improved gluconeogenic response to substrate when compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230334 TI - Influences of different methods of intramedullary femoral nailing on lung function in patients with multiple trauma. AB - We investigated the effects of primary (< or = 24 h) intramedullary femoral nailing on lung function and pulmonary hemodynamics in patients with multiple trauma. Two groups were separated: the group with primary femoral nailing with reaming (group RFN) was submitted to femoral nailing after reaming of the medullary canal; in the group with unreamed femoral nailing (group UFN) a small diameter solid nail was inserted without reaming. Lung function was assessed by oxygenation ratio (PaO2/FIO2), and pulmonary hemodynamics by intraoperative pulmonary artery catheter measurements. Central venous blood concentrations of elastase and the platelet count were determined during and 3 days after surgery. Lung function was stable in UFN patients (n = 14), but deteriorated in RFN patients (n = 17) from 353 +/- 24 (PaO2/FIO2 preoperatively) to 260 +/- 28 (PaO2/FIO2 postoperatively) (p < 0.05) and improved only after 48 hours. Pulmonary artery pressure (PAP) did not change during surgery in UFN patients; in RFN patients PAP increased from 27.2 +/- 3.1 mm Hg (preoperatively) to 36.3 +/- 4.1 mm Hg (p < 0.05) upon reaming and normalized 1 hour after insertion of the nail. Femoral nailing after reaming represents a potential risk with respect to lung function disturbances. This might trigger the development of adult respiratory distress syndrome (ARDS), especially in patients at extra risk of this complication (additional lung contusion, "borderline patient"). In these cases unreamed femoral nailing might offer an alternative by allowing primary intramedullary stabilization without the risk of adverse effects to the lung. PMID- 8230335 TI - Motor vehicle deaths: a rural epidemic. AB - OBJECTIVE: To determine the magnitude of the discrepancy in injury death rates between urban and rural counties and which types of injury deaths contribute most to this discrepancy. DESIGN: A review of Nebraska death certificates over the period 1985-1989 was undertaken. Counties were divided into four groups according to population. Group I: urban counties (n = 3); group II: counties with a town of greater than 10,000 (n = 9); group III: counties with a total population of greater than 10,000 (n = 19); group IV: counties with a total population of less than 10,000 (n = 62). Age-adjusted death rates for heart disease, cancer, cerebrovascular disease, pneumonia, and injury were tabulated. Injury deaths were further categorized by intentional injury (homicide, suicide), and unintentional injury (motor vehicle-related, falls, drownings, poisoning, farm machinery related, choking, firearms-related, fire-related and burns). INTERVENTIONS: None. RESULTS: Age-adjusted death rates per 100,000 population (with 95% confidence intervals) in group IV were lower than in group I for heart disease: 209 (193.9 224.1) vs. 227.4 (216.3-238.5); cancer: 135.9 (123.7-148.1) vs. 176.3 (166.6 186.0); cerebrovascular disease: 39.9 (33.3-46.5) vs. 44.6 (39.7-49.5); pneumonia: 19.6 (15.0-24.2) vs. 23.4 (19.8-27.0); and intentional injury deaths: 13.3 (9.5-17.0) vs. 15.1 (12.2-18.0). However, age-adjusted unintentional injury death rates were 54.2% higher in group IV than in group I: 42.7 (35.9-49.5) vs. 27.7 (23.8-31.6). Motor vehicle-related death rates were 93% higher: 23.3 (18.2 28.4) vs. 12.1 (9.5-14.7); and farm machinery-related deaths were 1250% higher: 2.7 (1.0-4.4) vs. 0.2 (-0.1-0.5). CONCLUSION: Age-adjusted unintentional injury death rates are higher in the rural counties of Nebraska, even though death rates for the four other leading causes of death (heart disease, cancer, cerebral vascular disease, and pneumonia) and intentional injury are lower. Although farm machinery-related deaths have the largest percentage difference between rural and urban counties, motor vehicle-related deaths are the major contributor to the unintentional injury death rate discrepancy in rural Nebraska. PMID- 8230336 TI - Hemorrhagic shock in endotoxin-resistant mice: improved survival unrelated to deficient production of tumor necrosis factor. AB - Although tumor necrosis factor (TNF) has been implicated in sepsis-induced mortality, its role in the pathophysiology of hemorrhagic shock (HS) remains ill defined. We studied three groups of acutely anesthetized mice undergoing HS to determine the role of TNF in HS mortality. Shock was initiated in each group after heparinization by arterial bleeding of 4 mL/100 g body weight followed by 12 mL/100 g body weight resuscitation with normal saline at 1 hour. The C3H/HeJ mice (n = 14), characterized by a genetic defect in macrophage production of TNF and other cytokines in response to endotoxin, were compared with the closely related C3H/HeN strain (n = 18), which do produce TNF. A second group of C3H/HeN mice were passively immunized to TNF by pretreatment with 2.5 mg/kg anti-murine TNF antibody (Ab) before HS. In contrast to the high TNF levels measured following HS in C3H/HeN controls, post-HS TNF was undetectable in C3H/HeJ mice. Five-day survival rate and survival time were significantly greater in C3H/HeJ mice when compared with C3H/HeN controls. Anti-TNF Ab pretreatment of C3H/HeN mice abolished the increase in TNF but did not improve survival. The data demonstrate a striking improvement in survival of TNF-deficient C3H/HeJ mice following severe HS. However, the improved survival does not appear to result from deficient TNF production, since Ab pretreatment did not decrease HS mortality. The improved survival in C3H/HeJ mice suggests that cytokines other than TNF may play a role in the pathophysiology of HS. PMID- 8230337 TI - Tube thoracostomy for occult pneumothorax: a prospective randomized study of its use. AB - Occult pneumothorax is defined as a pneumothorax that is detected by abdominal computed tomographic (CT) scanning, but not routine supine screening chest roentgenograms. Forty trauma patients with occult pneumothorax were prospectively randomized to management with tube thoracostomy (n = 19) or observation (n = 21) without regard to the possible need for positive pressure ventilation, to test the hypothesis that tube thoracostomy is unnecessary in this entity. Eight of the 21 patients observed had progression of their pneumothoraces on positive pressure ventilation, with three developing tension pneumothorax. None of the patients with tube thoracostomy suffered major complications as a result of the procedure. Hospital and ICU lengths of stay were not increased by tube thoracostomy. Patients with occult pneumothorax who require positive pressure ventilation should undergo tube thoracostomy. PMID- 8230338 TI - Evaluation of pregnant women after blunt injury. AB - A retrospective review of 125 pregnant women with blunt injuries admitted to a level I trauma center over a 35-month period was performed. The usefulness of three diagnostic tests, fetal ultrasound (US), external fetal monitoring (EFM), and Kleihauer-Betke (KB) tests in detecting fetal or pregnancy-associated complications was evaluated. The majority of women (77.6%) were involved in motor vehicle crashes and the mean injury Severity Score was low (4.7). The most common complications were premature uterine contractions (67%) and abruptio placentae (11%). When used together, EFM and US identified all complications. Moreover, all complications were manifest within 6 hours of admission. The KB tests had a sensitivity of 56%, a specificity of 71%, and an accuracy of 27%. We conclude that EFM and US are more useful in detecting fetal or pregnancy-associated complications after blunt injury. Monitoring can be limited to 6 hours if previous monitoring is normal. The KB test is of little use in the setting of acute trauma. PMID- 8230339 TI - Pattern of external birth trauma in southwestern Saudi Arabia. AB - The pattern and associations of 62 instances of external birth trauma (EBT) in 57 of 2222 consecutive hospital-born neonates were studied. Macrosomia and vaginal instrumental or breech delivery were identified as significant risk factors for EBT. In contrast to previous reports, subgaleal hemorrhage (SGH) was the predominant form of trauma (n = 33) and it was associated with vacuum application in 28 of 33 (85%) neonates. Brachial plexus palsy (BPP) (n = 10) occurred with the highest frequency (6.4%) after breech delivery. Fractures were uncommon. Multiple injuries (10 items in 5 babies) were mainly associated with vacuum extraction. The high rate of vacuum extraction (VE) and problems with its use including a high VE failure rate (30/242 = 12.4%) were considered contributory to the unusual pattern of EBT seen in the study. Regular audit of delivery practices, greater care in excluding cephalopelvic disproportion and judicious use of VE are recommended to reduce the morbidity from EBT. PMID- 8230340 TI - The frequency of bacteremia and fungemia following wound cleaning and excision in patients with large burns. AB - Twenty-eight patients with large burn injuries (mean total body surface area burned, 47.1%) who underwent 112 wound cleaning and staged early excision procedures were studied prospectively for bacteremia and fungemia induced by wound manipulation. The patients were given an aminoglycoside preoperatively, perioperatively, and postoperatively. Blood samples were obtained immediately before removal of dressings, after wound cleaning, after 30 minutes of surgery, at the end of each procedure, and 1 hour after surgery was completed. In a group of control patients blood samples were obtained immediately before the dressings were removed, after wound cleaning was completed, and 1 hour afterward. In a second group of control patients blood samples were obtained serially while the burn wounds remained undisturbed. None of the control patients received prophylactic aminoglycosides. Induced bacteremia or fungemia was documented in 50 instances of burn wound cleaning and excision; 31 cases of bacteremia or fungemia occurred after wound cleaning alone. Spontaneous bacteremia, i.e., that occurring in the absence of burn wound manipulation, was demonstrated in 3 of 18 blood culture series, whereas induced bacteremia was observed after 11 of the 17 burn wound cleaning procedures alone. The frequency of bacteremia tended to be higher for patients with inhalation injury than for those with no lung involvement. However, lung infection did not seem to account for many of the positive results of blood culture in this study. In contrast, burn wound infection contributed significantly to both spontaneous and induced bacteremia or fungemia. The micro organisms most frequently isolated were staphylococci (50%) and enterococci (15.1%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230341 TI - Effects of burn injury on class-specific B-cell population and immunoglobulin synthesis in mice. AB - Mice of the C57BL/6 strain were studied to investigate the effects of a 20% total body surface area full-thickness burn on class-specific immunoglobulin synthesis. Five days after injury single cell suspensions of spleen and mesenteric lymph node cells were made. The number of class-specific immunoglobulin-bearing cells were counted by flow cytometry using Texas-red-labelled antibody to class specific mouse immunoglobulin. The remaining cells were cultured with lipopolysaccharide for 4 days. The supernatant was tested for class-specific immunoglobulin by enzyme-linked immunosorbent assay (ELISA). There was no change in the number of class-specific IgG and IgA lymphocytes, but there was a significant increase in IgM lymphocytes. Synthesis of immunoglobulin by 10(5) lymphocytes was significantly increased in IgG and IgA cells, but decreased in IgM cells. Similar results were shown in spleen and mesenteric lymph node cells. Because the IgM response was much greater than others in the number of cells and immunoglobulin synthesis, we conclude that the decrease in responsiveness of IgM may contribute significantly to increased host susceptibility to infection after burn. PMID- 8230342 TI - A comparison of the effects of skeletal muscle injury and somatic afferent nerve stimulation on the response to hemorrhage in anesthetized pigs. AB - The effect of skeletal muscle injury (SMI) on cardiovascular and O2 transport responses to hemorrhage (HS) were examined in anesthetized pigs. Bilateral hindlimb muscle was injured 75 minutes before HS was started at a rate of 0.75 mL/min.kg until a total of 30 mL/kg (40% estimated total blood volume) had been removed. The reductions in cardiac index (CI), left ventricular stroke work, and oxygen delivery (Do2) and the increase in plasma lactate concentration following HS were exacerbated by SMI such that although oxygen consumption was maintained after HS it fell after SMI + HS. The deleterious effect of SMI on the response to HS was greater than that recorded previously following somatic brachial nerve stimulation (BNS). Thus, in order to achieve a given reduction in CI and Do2 or a rise in Shock Index (heart rate divided by systolic blood pressure) to approximately 3, a blood loss of 40% was needed after HS; this was reduced to 36% by the addition of BNS, whereas a loss of only 29% was needed when SMI was introduced. The mechanism of the deleterious effect of SMI is unclear although a change in the distribution of regional blood flow and a rise in the critical oxygen delivery may be implicated. PMID- 8230343 TI - Laparoscopic injection of fibrin glue to arrest intraparenchymal abdominal hemorrhage: an experimental study. AB - The laparoscope offers a novel avenue for the diagnosis of intra-abdominal injury and the use of fibrin glue (FG) as a treatment for hemorrhage in trauma patients. This study was undertaken to assess the practicality and effectiveness of FG injection under laparoscopic direction to arrest hemorrhage in solid viscera. Twenty dogs were randomized into a control group (CG) and a treatment group (TG). All animals underwent laparotomy to surgically induce uniform injuries to the hepatic and splenic parenchyma. The TG animals (n = 12) were allowed to hemorrhage for 30 minutes. The injuries were then visualized and FG injected intraparenchymally under laparoscopic direction. The average duration of the procedure was 25 minutes (range, 15-50). No hemostatic interventions were performed on the CG animals (n = 8). Mortality in the CG was 63% (5 of 8); there were no deaths in TG animals prior to sacrifice. Necropsy of TG animals revealed progressively healing hepatic and splenic injuries with no gross evidence of pulmonary FG emboli, intraparenchymal microemboli, or increased adhesion formation. No other complications were noted. This study demonstrates that hemorrhage from the liver and spleen can be successfully controlled using the laparoscope to direct the intraparenchymal injection of FG. In this experimental model, the procedure can be performed expeditiously. It is associated with reduction of mortality to zero when compared with controls. No complications associated with laparoscopy or FG injection were recognized. This technique may have potential for application in the management of stable patients who manifest evidence of intraperitoneal hemorrhage as a result of solid organ injury. PMID- 8230344 TI - Increased rates of complications in patients with severe ankle fractures following interinstitutional transfers. AB - A retrospective review of 80 consecutive patients treated at a county hospital for ankle fracture/dislocations (initial displacement > 10 mm) was undertaken. Seventy-seven patients fulfilled the requirements for documentation and follow up. Eighteen patients were transferred from another facility and 59 patients were seen at our institution primarily. The transfer and no-transfer groups were similar in age, sex, follow-up, type of ankle fracture, and number of concomitant injuries. The no-transfer group had a slightly higher Injury Severity Score and incidence of active medical problems. Patients were transferred to this public hospital for financial considerations in 89% of the cases. The patients seen in transfer had a higher rate of overall complications (67% vs. 26%; p = 0.0001), including deep infections (11% vs. 1.7%) and major reduction failures (17% vs. 7%). Higher rates of complications were also found in patients with delayed time to operation, psychiatric or substance abuse problems, and those with skin abrasions or blisters. Analysis of variance indicated that transfer status was the strongest predictor of complications. Hospital stays were also longer in the transferred group. PMID- 8230345 TI - Closed interlocked nailing in comminuted and segmental femoral shaft fractures. AB - One hundred eight femoral shaft fractures in one hundred six adults were treated by closed intramedullary interlocked nailing. Ninety-two fractures were severely comminuted and 16 fractures were segmental. The procedures were done with the patients in the supine position. A modification of the cylinder guide for fixation of the distal screws is described. All the fractures healed in an average of 18 weeks (range, 12-24 weeks). Dynamization was performed in 15 fractures. There were two postoperative complications, which did not ultimately affect the clinical results. No infections or delayed unions were observed. Closed intramedullary interlocked nailing seems to be the treatment of choice for comminuted and segmental fractures of the femoral shaft provided that all the technical details are followed by the surgeon. PMID- 8230346 TI - Cortical bone reactions at the interface of external fixation half-pins under different loading conditions. AB - The pin-bone interface is the weakest link in the mechanical stability of external skeletal fixation. In this investigation, a canine model was used to characterize the nature of cortical bone reactions at the pin-bone interface. Unilateral external fixators were applied to the tibiae of 61 dogs using six tapered cortical half-pins. The pins were inserted after predrilling both cortices, and pin insertion torque was measured. A transverse or oblique osteotomy was performed in each tibia and stabilized under different gap conditions. Unrestricted weight bearing was allowed immediately after surgery. Sequential functional evaluations of the animals and biweekly roentgenograms of the pin tracks were done. Pin removal torques were recorded at the time the animals were killed at different time periods. Pin tracks were analyzed using quantitative tetracycline histomorphometry and microradiography. The results showed that cortical bone undergoes extensive creeping substitution around external fixation half-pins. New bone accounted for approximately 43% of the intracortical space along the pin track, and cortical bone porosity showed a fourfold increase compared with intact bone value. This cortical bone remodeling resulted in a time-related decrease of pin removal torque (p < 0.001). In inherently unstable oblique osteotomies, and less in stable rigidly fixed transverse osteotomies, immediate postoperative weight bearing caused bone thread resorption and adverse cortical bone remodeling at the entry cortex of external fixation half-pins. The unicortical loosening of half-pins that became evident during the first month of fixation obviously represents a consequence of micromotion and local bone yielding failure caused by high dynamic stresses of the pin-bone interface. Effective precautions should be taken to reduce such stresses. PMID- 8230347 TI - Facts versus values: why legislators vote against injury control laws. AB - BACKGROUND: Control of motor vehicle-related injuries depends upon passage of mandatory safety belt and other injury control laws. Unfortunately, state legislators often oppose these laws. METHODS: In 1988, a 62-item questionnaire was mailed to the 97 Colorado legislators who voted on a 1987 safety belt law to identify factors (knowledge, experiences, attitudes, and beliefs) associated with "yes" and "no" votes. To test for associations between these attributes and the legislators' recorded votes, odds ratios (OR) and 95% confidence intervals (CIs) were calculated. A stepwise logistic regression identified independent predictors of "vote." RESULTS: Fifty-three (55%) of the legislators responded. Responders and nonresponders were demographically similar. "Vote" was not associated with age; sex; having young children in the family; perceived injury risk; recent traffic tickets; family or personal crash experience; or knowledge of the fatality risk reductions attributable to wearing safety belts. Ninety-six percent of the legislators knew that safety belts reduce the risk of death and 87% believed a safety belt law would save lives. The strongest predictors of a "yes" vote were impression that constituents favored the law (OR = 31, CI 95 = 3.5, 270); belief that a mandatory safety belt law will save lives (OR = 20, CI 95 = 2.1, 203); and "extreme" importance paid in the voting decision to effectiveness of the law in reducing deaths (OR = 19, CI 95 = 3.5, 107). Legislators who considered restrictions on individual freedoms an "extremely" important decision criterion were 43 times (CI 95 = 7, 267) more likely to vote "no." In the logistic model only extreme importance assigned to individual freedoms (beta = 3.7; OR = .025; p = 0.002) and policy effectiveness (beta = +3.1; OR = 22; p = 0.01) predicted "vote." The logistic model correctly predicted 90% of legislators' votes. CONCLUSIONS: In this study the strongest predictors of voting behavior were concern for individual freedoms, perceived constituents' support and attention paid to policy effectiveness. Those seeking to persuade legislators to vote for mandatory safety belt laws must pay attention to attitudes and values in addition to scientific facts. PMID- 8230348 TI - Neck injuries among belted and unbelted occupants of the front seat of cars. AB - This article, part of a recently completed research project on safety belts, presents results on neck injuries. A total of 3927 injured front-seat occupants (drivers and passengers) involved in two-car crashes were studied. Among them, 725 sustained neck sprains (ICD-9 code 847.0); some of them may have sustained other injuries as well. The more serious injuries to the cervical spine were more prevalent among the unbelted occupants. Neck sprains were relatively more numerous among belted occupants compared with unbelted ones, with a relative risk estimate of 1.68. Similar results hold also for subsets of the data on different types of collisions; the relative risks ranged from 1.39 to 2.42. A log linear model was constructed for the odds ratio (neck sprain vs. no neck sprain) taking into account the following factors: (1) seatbelt use, (2) direction of impact, (3) authorized speed limit, and (4) vehicle weight. The resulting relative risk estimate (belted vs. unbelted) became 1.58. The results raise questions about seatbelts and their protection against neck sprains. PMID- 8230349 TI - The effects of continuous operation in a chemical protective ensemble on the performance of medical tasks in trauma management. AB - Treating casualties in a chemically hazardous environment constitutes a unique problem. Physical protection of the medical personnel may impair their performance and potentially affect patients' prognoses. The present study examined the effect of prolonged physical protection on the accomplishment of medical tasks related to trauma management. Sixty one emergency medical technicians, acclimatized to operating in protective gear, underwent four rounds of testing during eight hours of continuously wearing either a chemical protective suit or regular fatigues. The quality of the designated medical tasks, including sterility, was maintained throughout the study. A significant reduction in speed of performance was noted (approximately 30% slowing, p < 0.0001 in multivariate analysis) because of protective clothing. There was no additional decrement in performance following a prolonged stay in the protective gear. We conclude that in a chemically contaminated area, fully protected medical personnel are capable of treating trauma patients reasonably well, and for a relatively long period of time. The importance of pretraining and proper instruction is emphasized. PMID- 8230350 TI - Failure of jugular lactate determinations to disclose cerebral ischemia in posttraumatic cerebral infarction: case report. AB - A case of acute closed brain trauma is presented in which the patient developed a right middle cerebral artery infarction, while frequent measurements of the arteriojugular lactate difference (AVDL) remained normal, failing to disclose ischemia. To our knowledge, this is the first report on sequential AVDL measurements during evolving cerebral infarction. PMID- 8230351 TI - Intramedullary stabilization of a bowing fracture of the forearm with Ender's nails: case report. AB - A case history of a young man with a bowing fracture of the forearm is presented. The fracture was reduced and stabilized using shortened Ender's nails. Both soft tissues and fractures healed uneventfully and complete restoration of function resulted. PMID- 8230352 TI - Air-to-surface missile wound of the thorax reconstructed with a polytetrafluoroethylene patch: case report. AB - A method of reconstructing a chest wall defect following non-guided air-to surface missile injury is described. The wall defect was simply closed with a polytetrafluoroethylene patch. The high risk of infection following use of the described method is well recognized. No infection occurred in this case. It could be stated that under the exigencies of the moment the use of prosthetic material to reconstruct the chest is not invariably followed by infection. However, when the patient is stable this latter problem can then be addressed by a variety of standard methods. It is concluded that the described method is particularly suitable in war circumstances, when it is usually necessary to attend to a large number of patients in a short period of time. PMID- 8230353 TI - Spontaneous voiding of a bullet after a gunshot wound to the bladder: case report. PMID- 8230354 TI - [DNA methylation of monohalogenated methanes of F344 rats]. AB - Monohalogenated methanes (methyl chloride, methyl bromide and methyl iodide) are mutagenic and carcinogenic. The possible mechanism of these effects, DNA methylation, was studied. DNA adducts from organs of F344 rats exposed to these chemicals were separated and identified with high performance liquid chromatography (HPLC) and gas-chromatography/mass-spectrometry (GC/MS). DNA adducts, 7-methylguanine (7-MeG) and O6-Methylguanine(0(6)-MeG), incorporation of 14C into de novo synthesis of nucleobases could be observed in enzymatic DNA hydrolysates by HPLC and determination of the radioactivity in the fractions. The formation of DNA adducts in the studied organs was only quantitatively different. The formation of O6-MeG was further proved by analysing the acidic hydrolysates using HPLC with non-radioactive O6-MeG as internal standard. 7-MeG and 3-MeA were identified with GC/MS analysis. PMID- 8230355 TI - Immunological status of patients with obstructive jaundice and immunostimulatory effect of arginine. AB - Sepsis is a major factor in the high mortality and morbidity following diagnostic and therapeutic procedures in patients with obstructive jaundice. The reasons for this increased susceptibility to infection are not fully understood. We therefore observed prospectively changes in immunological status of patients with obstructive jaundice in the perioperative period and studied immunological effects of perioperative arginine therapy. The results showed that there was a significant reduction in interleukin 2 (IL-2) production, interleukin 2 receptor (IL-2R) expression and lymphocyte response to phytohemagglutinin (PHA) mitogen in patients with obstructive jaundice compared with normal controls. After operation, the immune suppression in patients with obstructive jaundice was more significant. Arginine is a known T lymphocyte stimulator. Perioperative supplement with arginine significantly enhanced the immune function of patients with obstructive jaundice, the mechanism being related to increased IL-2 production and IL-2R expression. PMID- 8230356 TI - [The change of T lymphocyte subpopulations, interleukin-2 receptor and HLA-DR antigen in lupus nephritis patients]. AB - In this study peripheral blood mononuclear cells were tested for T lymphocyte subpopulations, interleukin-2 receptor (IL-2R) and HLA-DR antigen by monoclonal antibodies in 14 LN patients. In comparison with normal controls, 14 LN patients showed a decrease of T4 lymphocyte (0.01 < P < 0.05) and increase of T8 cells (P < 0.01) with reduction of T4/T8 ratio (P < 0.01). The expression of HLA-DR cells increased (P < 0.01), the expression of IL-2R activated by PHA significantly decreased (P < 0.01), while nonactivated IL-2R expression significantly increased (P < 0.01). T4/T8 ratio was significantly correlated with activated IL-2R expression (P < 0.01, r = 0.6599). The results indicated that there was a immunological defect in LN patients. The pathogenesis of LN might be heterogeneous. PMID- 8230357 TI - Levonorgestrel levels in serum and uterine flushings in women bearing LNG-Cu-IUD. AB - Radioimmunoassay was employed to measure levonorgestrel levels in serum and uterine flushings in women using intrauterine device releasing both copper ions and levonorgestrel (LNG-Cu-IUD) after one-year insertion. In the meanwhile physical characteristics of cervical mucus have been observed in women using LNG Cu-IUD and compared with those using Tcu-220c-IUD. The mean concentrations of levonorgestrel in serum and uterine flushings (5 ml) were 84.53 pg/ml and 266.93 pg/ml respectively. In LNG-Cu-IUD group, the cervical mucus lacking in ferny crystals had a viscosity significantly higher than that in Tcu-220c-IUD group, suggesting that the local high concentration of levonorgestrel changed the environment of the uterine cavity and the cervical mucus status. PMID- 8230358 TI - Impaired antigen-presenting capability of monocytes correlated with their decreased expression of HLA-II antigens in patients with myeloid leukemia. AB - Antigen-presenting capability and expression of HLA-II antigens on peripheral blood monocytes were analyzed by isotope incorporation technique and indirect immunofluorescence assay with monoclonal antibodies Tu 22, Tu 36 and anti-Leu-M2 in patients with acute and chronic myeloid leukemias (AML, CML). All patients (17 AML and 13 CML) demonstrated impaired antigen-presenting capability of monocytes (P < 0.001), when compared with simultaneously studied controls, which were HLA identical normal siblings of the patients. These patients also showed a markedly decreased proportion of MAC-120- and HLA-DQ-positive monocytes as compared with the controls (P < 0.001), while the percentage of HLA-DR-positive monocytes in the patients was similar to that observed in the controls (P > 0.05). These findings suggested, therefore, that impaired antigen-presenting capability of monocytes correlated with their decreased expression of HLA-DQ in patients with acute and chronic myeloid leukemias. PMID- 8230359 TI - [Study on the relationship between most cells and tumor growth speed during hepatocarcinogenesis in the rats]. AB - The nucleolar organizer regions (NORs) of normal rat hepatocytes and hepatocytes in the precancerous hyperplastic nodules as well as the cells of hepatocellular carcinoma induced by Nitrosomorpholine in rats were observed with aid of AgNOR staining and electron microscopy. The observation revealed that the number of Ag granules in the NORs of carcinoma cells was significantly more than that in the hyperplastic and normal hepatocytes. The ultrastructural observation of the nucleoli of interphase nuclei showed just the same result as the AgNOR-staining. The alcian blue-PAS-ABC combined staining revealed a very marked infiltration of mast cells (MC) within the carcinoma tissue and in its surroundings in 4 animals, while other 5 animals showed only a slight or no MC infiltration in the surroundings of carcinoma tissue. The observation of the phenotype of MC revealed that only one of the animals with carcinoma showed a relatively marked infiltration of both mucosa mast cells (MMC) and connective tissue mast cells (CTMC), while all other rats showed mainly an infiltration of CTMC. The Ag granules in the NORs of carcinoma cells of the carcinomas with more MC infiltration were less than that of carcinomas with only slight or without MC infiltration. The difference was statistically significant. The significance of above mentioned results was discussed and a relationship between MC and prognosis of liver carcinoma was suggested. PMID- 8230360 TI - Effects of GAL10-SUC2 promoter combinations on SUC2 gene expression in S. cerevisiae. AB - The plasmid series YEP51 delta n bearing GAL10-SUC2 promoter combinations were constructed by inserting SUC2 gene with different upstream deletions downstream GAL10 promoter on YEP51. After transforming yeast cells S. cerevisiae, the invertases expressed by each of the transformants were measured and analysed by means of PAGE. The results showed that: 1) The SUC2 gene with upstream deletion to at -636bp expressed high level glycosylated form of invertase under glucose derepression, while SUC2 gene with more extensive deletions to -223 bp or more lost its response to glucose derepression; 2) Each part of GAL10-SUC2 promoter combination acted almost independently. All of the combinations showed no apparent coordinated promoter function under our experimental conditions; 3) Sequences between -89bp and -41bp of SUC2 upstream region are responsible for constitutive expression of nonglycosylated invertase. The two tracts of poly (dA dT) of this region may serve as promoter elements. PMID- 8230361 TI - The regulatory effect of substances released from porcine bronchial epithelial cells on hypoxic pulmonary vasoconstriction. AB - In this study, porcine pulmonary artery segments (intact or denuded) and bronchial segments (with or without epithelium) were tested in organ bath. Hypoxia caused a bronchial epithelium dependent relaxation of intact or denuded pulmonary artery preconstricted with phenylephrine. Constriction in intact pulmonary artery coated with epithelium denuded bronchus was observed during hypoxia. The bronchial epithelium-dependent relaxation could be inhibited by indomethacin but not be blocked by the following agents: atropine, propranolol, gossypol, methylene blue and chlorpheniramine. The results suggested that HPV was endothelium-dependent. Hypoxia could cause the production of an epithelium derived relaxing factor (EpDRF), which acted directly on smooth muscle. The effect of EpDRF was not mediated by pulmonary endothelium, but might be related to arachidonic acid cyclooxygenase pathway. PMID- 8230362 TI - Effect of hypoxia on release of vasoactive substances from cultured pulmonary arterial and aortic endothelial cells. AB - The effect of hypoxia on the production and release of vasoactive substances from endothelial cells of pulmonary artery (PAECs) and aorta (AECs) was studied. The results indicated that the overall effect of the long half-life vasoactive substances released from PAECs and AECs was vasoconstrictive. The long half-life lipid-soluble substances produced by PAECs and AECs were vasodilative, and did not change in hypoxia. However, the long half-life water-soluble heat-unstable and heat-stable ones were vasoconstrictive. Hypoxia could reduce the release of the former and promote that of the latter which might be peptides. The PAECs could release specific long half-life mediator which was pulmonary artery constrictive, water-soluble, heat-unstable, and not related to hypoxia. Hypoxia inhibited the production of PGI2, a short half-life vasodilator, in PAECs, but not in AECs. PMID- 8230363 TI - Electrophysiological and relevant morphological properties of neurons in toad dorsal root ganglion. AB - Intracellular recordings were performed on isolated preparations of toad dorsal root ganglion (DRG). Of the 87 neurons examined, 83 were of type A cells and 4 of type C cells. The conduction velocity (CV) of type A cells ranged from 2.55 to 35.19 m/s with a mean of 11.18 +/- 0.78 m/s (mean +/- s mean), while that of type C cells ranged from 1.34 to 2.43 m/s with a mean of 2.11 +/- 0.22 m/s (mean +/- s mean). On the basis of the characteristics of Ap configuration, these 87 cells can be classified into three groups: H neuron (24/87); F neuron (50/87) and F-H neuron (13/87). The Sizes of DRG cells were examined by measuring of HRP labelled cells in part of the experiment. By comparing the size of F and H cell with their CV, we proposed a revised view which is against the public opinion concerning the relation between the size and CV of DRG cells. PMID- 8230364 TI - The effects on basal anterior pituitary hormone concentrations by cranial irradiation in patients with nasopharyngeal carcinoma. AB - Hypopituitarism occurring in patients who received external cranial radiation therapy (RT) have been increasingly recognized. Fourteen healthy patients were collected as control Group I, and 25 patients with nasopharyngeal carcinoma (NPC) were enrolled as Group II. Another 18 NPC patients were enrolled as Group III. Blood was sampled from Group I, Group IIB (before RT), Group IIA (one week after RT) and Group IIIA (1 to 1.5 years after RT). The basal samples were taken for growth hormone (GH), adrenocorticotropic hormone (ACTH), cortisol, follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), thyrotropin (TSH), triiodothyronine (T3) and thyroxine (T4). The results showed the T3 level was statistically low in Group IIB compared with Group I. The serum GH and FSH (male only) levels were higher and with statistical significance in Group IIA compared Group IIB. Both GH and ACTH levels decreased and PRL increased with statistical significance in Group IIIA compared with Group IIA. The ACTH decreased with statistical significance in Group IIIA compared with Group IIB. Clinical damage to the pituitary is usually manifested months to years after RT. Therefore, a careful exclusion of these glands from radiation treatment fields is recommended whenever possible. PMID- 8230365 TI - Xe-133 muscle blood flow in polymyositis and dermatomyositis. AB - The histologic picture of polymyositis or dermatomyositis is characterized by muscle fiber degeneration, which is assumed to be caused by a circulatory abnormality. To verify this assumption, we measured the muscle blood flow in patients with polymyositis (or dermatomyositis) and in normal subjects using the inert gas Xe-133. Sixteen patients and 30 age-matched normal volunteers were studied. Muscle blood flow was measured with Xe-133 dissolved in isotonic saline as described by Lassen et al. (1964) and Pozderac et al. (1975). Data were acquired in the frame mode after intramuscular injection (anterior tibial muscle) of 0.3 mCi of Xe-133 in 0.2ml of saline for 20 min. A computer routine was used to calculate the clearance half time (T 1/2) in min and flow rate (Q) in ml/100 gm tissue/min. The student's t test was used to test statistical significance. Our preliminary results suggest: 1. The muscle blood flow in patients with polymyositis and dermatomyositis was significantly lower than the corresponding normal control levels. 2. This findings may be due to decreased in muscle capillary 3. Further studies are warranted to obtain a conclusive understanding of the pathogenesis of polymyositis and dermatomyositis. PMID- 8230366 TI - [The mechanism of the pineal gland in inhibiting sexual receptivity of female rats treated with monosodium-L-glutamate: (III). Does it concern with the function of the pituitary gland?]. AB - To elucidate whether pituitary function participates in the effect of the pineal gland on sexual receptivity, monosodium L-glutamate (MSG) was used as a neurotoxin to induce hypogonadal status. Long-Evans rats were divided into four groups: (1) normal control (C), (2) pinealectomized (Px), (3) MSG-treated (MSG) and (4) pinealectomized MSG-treated (Px-MSG). Pinealectomy was performed at six weeks of age. In the first part of the experiment, the sexual receptivity was estimated at the age of 2.5 months by lordosis quotient (LQ). The result indicates that the decline of receptivity by neonatal MSG treatment can be significantly improved by pinealectomy. In the second part of the experiment, the effect of pinealectomy on pituitary function was examined by two tests including (1) post-castrational LH rise and (2) pituitary response to LHRH. Ovariectomy was performed at the age of 2.5 months. Four weeks later three consecutive blood samples were collected at 10 minute intervals for LH radioimmunoassay. Then, three doses of LHRH (100 ng, 250 ng and 500 ng/100 g of body weight) were administered separately at two-week intervals, serum samples were taken before as well as 15 and 60 minutes after LHRH administration. The results showed that there was no significant difference in serum LH levels between Px and control rats after ovariectomy. The LHRH-evoked LH elevation in Px-MSG rats was just the same as that of the MSG rats, although the LH level in MSG rats was lower than in the control rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230367 TI - ["Walkman music" during epidural anesthesia]. AB - The sedating effect and influence of walkman music on 120 patients, that were randomly divided into a music and non-music group, who had received epidural anesthesia were investigated. It was found that significantly fewer patients in the music group felt anxious during surgery (P = 0.049). Meanwhile, heart rate and mean arterial pressure of the non-music group remained at a higher level, resulting in higher heart loading. Furthermore, patients listening to music had a significantly smaller need of sedatives (P = 0.001). With the purpose of reducing the consumption of sedatives, and offering a better anesthetic environment, we recommend the use of music during regional anesthesia. PMID- 8230368 TI - [Congenital nasopharyngeal teratoma: report of a case and review of the literature]. AB - Teratomas are the most common congenital tumors, but teratomas of the nasopharynx are rare and seen almost exclusively in infants, usually in neonates. An unusual case of a neonate with respiratory distress is presented and a nasopharyngeal mass protruding into the oral cavity. After successful removal of the mass, pathological examination revealed a mature teratoma. The management and differential diagnosis are discussed, accompanied by a review of the literature. PMID- 8230369 TI - The management of advanced subglottic carcinoma with stomal invasion: a case report. AB - Primary subglottic carcinoma is rare. The initial stage is usually asymptomatic and the carcinoma is advanced when the symptoms are in evidence. Because the trachea and thyroid gland adjoin to the subglottic area, the tumor invades them along with the superior mediastinum and lung by the way of paratracheal lymph drainage. To resuscitate a severe airway obstruction, an emergency tracheotomy is required, which, however, results in a higher frequency of stomal recurrence later. We treated a case with advanced subglottic carcinoma and performed extended radical neck dissection with the resection of total larynx, thyroid gland, and involved trachea with peristomal skin en bloc. In order to completely remove the paratracheal and superior mediastinal lymph nodes, the sternal manubrium and its adjoining clavicles and ribs were cut off. This was necessary to optimize the field of vision, and to prevent injury to the great vessels and pleura. Because of its availability, its thickness, and its skin paddle, the pectoralis major myocutaneous flap was formed to the skin defect and to protect the great vessels. Postoperative stoma was at the sternal level and did not compress the great vessels. Parathyroid gland, after chopping, was reimplanted into muscle tissue to prevent hypocalcemia. No more postoperative radiotherapy or chemotherapy was necessary because the lymph nodes were spared. There were no postoperative complications and the patient was discharged 2 weeks later. No recurrence occurred in the 28 months follow-up period. PMID- 8230370 TI - Enterolith ileus as a complication of duodenal diverticulosis--one case report and review of the literature. AB - Duodenal diverticulosis is not a rare condition. Usually of little clinical significance, it can produce a variety of disorders such as malabsorption, hemorrhage, diverticulitis, and obstruction. The rarest complication appears to be enterolith formation and obstruction. The case presented is a 70-year-old woman with the chief complaints of intermittent abdominal pain and vomiting. At laparotomy, duodenal diverticulitis and one enterolith obstructing the distal ileum were found. The literature review presents the other 26 cases with small bowel obstruction due to an enterolith formed within a small bowel diverticulum. The diagnosis can be established only by documenting the normalcy of the gallbladder and the presence of duodenal or jejunal diverticula. PMID- 8230371 TI - Registration and display of multimodal images: applications in the extracranial head and neck region. AB - Image registration is a correlation procedure that allows the matching of images obtained by different imaging modalities, both structural and functional. Once registered, they can be combined in a single "fused" image. This technique can be used to improve the interpretation and the quantification of relatively low resolution images obtained in nuclear medicine or to achieve a better diagnosis and treatment planning when high resolution anatomical images are registered. Matching images obtained by different imaging modalities is a major challenge and the various registration techniques are briefly described. The extracranial head and neck region is a challenging field for image registration. A preliminary retrospective experience is presented emphasizing the technique feasibility in a routine clinical context. 2-D and 3-D approaches were used. The requirements for transferring registration technology from the image processing laboratory to clinical practice are discussed, and the contribution of this method to medical interdisciplinary communication is emphasized. PMID- 8230372 TI - MR imaging with Gd-DTPA in lesions of the head and neck. AB - The diagnostic value of MR in the head and neck with special emphasis on the contrast medium Gd-DTPA will be demonstrated. A total of 1,260 patients underwent MR with different pulse sequences plain and with Gd-DTPA. The sequences were analyzed and compared with CT. Due to the different pattern of enhancement the contrast medium helped in differentiating malignant and benign tumors from other processes like cysts and inflammatory changes. Except lesions with small bony erosion and inflammatory changes of the salivary glands MR with Gd-DTPA proved to be diagnostically advantageous in all cases. Gd-DTPA improved the delineation of tumor margins. MR in combination with Gd-DTPA offers a lot of new advantages for diagnosing lesions of the head and neck. However, due to the prolonged examination time and the lack of noninvasiveness the use of Gd-DTPA has to be considered critical for every single area of the head and neck. PMID- 8230373 TI - The relevance of positron emission tomography for the diagnosis and treatment of head and neck tumors. AB - A total of 48 patients with squamous cell carcinomas (SCC) of the head and neck were examined with positron emission tomography (PET) and 18Fluor-deoxyglucose (FDG). In 25 cases the FDG-PET-data were compared with flow cytometric data of the lesions to investigate the value of PET for the noninvasive determination of a tumor's proliferative activity. The quantitative analysis of the radioactivity concentrations revealed two groups of tumors with a different FDG-uptake. A correlation was seen between the proliferative index and the FDG-uptake with an r value of 0.64 in the lower uptake group and an r-value of 0.81 in the higher uptake group. There was no correlation between any clinicopathological or morphological parameters of the tumors and the FDG-uptake. It is suggested that the different FDG uptake may correspond to differences at the molecular level. The PET-evaluation of therapeutic effects in patients with advanced SCC of the head and neck revealed different posttherapeutic changes of the metabolism in tumors and in lymph node metastases. Tumors were more sensitive to therapy than lymph node metastases. There was a high correlation between the growth rate and the change in FDG uptake with a different regression function for tumors and for lymph nodes. The data demonstrate the usefulness of PET and FDG for the evaluation of early therapeutic effects. Therefore PET can be applied for the treatment planning in patients with malignant tumors of the head and neck. PMID- 8230374 TI - MRI and CT in the evaluation of acquired and congenital cholesteatomas of the temporal bone. AB - Acquired (secondary) cholesteatomas of the middle ear and mastoid are usually a complication of chronic otomastoiditis and are often accompanied by infection from the outset, and their contents show evidence of some inflammatory reaction. Congenital (primary) cholesteatomas of the temporal bone are due to epithelial rest of embryonal origin. There are many sites of occurrence of congenital cholesteatomas (epidermoids) within the temporal bone: (1) middle ear, (2) mastoid, (3) middle ear and mastoid, (4) petrous bone, (5) the squama, and (6) within the tympanic membrane. Intradural (cisternal) congenital cholesteatomas are another type of cholesteatomas that often involve the cerebellopontine angle (CPA) region and cause varying degrees of cochlear and vestibular symptoms and signs. In this paper, we stress the role of magnetic resonance imaging (MRI) and computed tomography (CT) in the evaluation of acquired and congenital cholesteatomas of the temporal bone. It is our opinion that CT remains the study of choice for cholesteatomas of the middle ear cleft. MRI is superior to CT for the evaluations of infected cholesteatomas, petrous apex, and CPA cholesteatomas, as well as for the majority evaluation of cholesteatomatous involvement of the facial nerve, membranous labyrinth, and intracranial structures. PMID- 8230375 TI - Temporal bone fractures: high resolution CT. AB - Apart from midfacial traumas, laterobasal fractures are the most common skull base injuries the ENT surgeon has to deal with. These fractures often appear in combination with more or less extensive further skull or brain injuries. In these cases interdisciplinary co-operation with the neurosurgeon is necessary. Today, development of high resolution computed tomography makes it possible to define priorities in treatment of these patients exactly. PMID- 8230376 TI - CT imaging in the diagnosis and treatment of sinus disease: a partnership between the radiologist and the otolaryngologist. AB - We consider that the nasal cavity and the paranasal sinuses cannot be declared completely normal without a normal CT scan. While CT scans are clearly useful in the evaluation of patients with recurrent sinusitis, it is important to emphasize that they should not be employed exclusively for diagnosis. Nasal endoscopy and clinical evaluation still form the basis for the diagnosis of chronic and recurring sinusitis. The CT technique described should be used only to supplement the clinical data obtained during history-taking and the rhinoscopic/endoscopic examination and is essential before functional endoscopic sinus surgery. The complexity of the diagnostic process is simplified to patient benefit when the radiologist and the otolaryngologist form a functional interactive partnership. PMID- 8230377 TI - Three-dimensional CT reconstruction and volume measurements of the pharyngeal airway before and after maxillofacial surgery in obstructive sleep apnea. AB - We believe that maxillofacial surgery is a successful procedure for treatment of sleep apnea, provided the patients are fully assessed pre-operatively with nocturnal polysomnography and pharyngeal airway imaging. Long follow-up is also necessary to evaluate success. PMID- 8230378 TI - Imaging evaluation of laryngotracheal stenosis. AB - Laryngotracheal stenosis (LTS) may afflict patients in all stages of life. Congenital subglottic stenosis is the third most common upper airway anomaly encountered in infants and neonates after laryngomalacia and vocal cord paralysis or fixation. It is a complex condition that demands arduous, often frustrating, efforts at all stages of management. Prevention of LTS is the most effective management of this problem. This paper will focus on the role of diagnostic radiology in the management of LTS. PMID- 8230379 TI - Deglutition after supra-glottic laryngectomy. AB - This study of swallowing after a horizontal supra-glottic laryngectomy proposes to analyze the frequency of long-term post-operative complications, their mechanisms of onset, as well as their compensatory mechanisms. It rests on the study of three groups of patients: a retrospective series of 74 charts, a group of 14 patients studied at a distance from the intervention, and a group of 15 patients who have been entered in a prospective study concerning the post operative care of partial laryngectomies. For these two groups, the swallowing work-up consisted of, besides a clinical evaluation, a video-laryngoscopy and a video-radioscopy. Knowledge of the compensatory possibilities as a function of the specifics of the surgical technique, and their evaluation for each patient with video-radioscopy allows for an adapted care protocol. PMID- 8230380 TI - CT and MR evaluation of laryngeal carcinomas. AB - A prospective and comparative tomography (CT)/magnetic resonance imaging (MRI) study on 110 patients with endoscopically examined and histologically proven laryngeal malignancy is presented. Post-operative pathological and intraradiological (CT vs. MRI) correlation are established. We conclude that MRI is the method of choice for staging laryngeal malignancies. MRI can also be used for follow-up (recurrence after surgery, evolution under radiation therapy). PMID- 8230381 TI - Incidence and types of non-palpable thyroid nodules in thyroids removed for palpable disease. AB - The purpose of this study was to determine the number, size and types of non palpable and ultrasound undetected thyroid nodules in thyroid glands removed for a single palpable nodule. Twenty patients undergoing partial (hemi- and subtotal) thyroidectomy and 40 patients having a total thyroidectomy were analyzed. Patients had no more than one palpable nodule, no cervical adenopathy and no history of previous thyroid surgery. An ultrasound was performed routinely within four months of surgery. Forty-six nodules were detected by final pathology in the partial thyroidectomy group but only 42% were detected by palpation and 59% by ultrasound. The median diameter of non-palpable and ultrasound undetected partial thyroidectomy nodules was 0.5 cm and all were benign except for one case of lymphoma. Pathology detected 186 nodules in the total thyroidectomy group although palpation only detected 22% of these and ultrasound 42%. The median diameters of the benign and malignant non-palpable nodules were 0.4 cm and 0.25 cm respectively. Most of the undetected nodules in the total thyroidectomy group were benign but there were 20 non-palpable and 18 ultrasound undetected papillary carcinomas. PMID- 8230382 TI - Pre-operative localization of 25 consecutive parathyroid adenomas: a prospective imaging/surgical correlative study. AB - The successful diagnosis and surgical treatment of primary hyperparathyroidism due to parathyroid adenoma benefits significantly, in our experience, from a process of pre-operative imaging localization of the parathyroid adenoma. This prospective study evaluates a window of 25 consecutive patients who underwent pre operative imaging localization prior to successful unilateral parathyroidectomy for parathyroid adenoma. All parathyroid adenomas were successfully localized by imaging, and subsequently documented photographically in surgical correlation, and pathologically confirmed. All patients were cured biochemically. Ultrasound accurately localized 92% of adenomas (100% in the neck and extrathyroidal) while radionuclide subtraction scanning identified 60% of a smaller subset. Both DSA and CT were successful in the two cases utilized, and MRI demonstrated four of five adenomas. The high yield of these pre-operative localization studies should make them an important consideration in the routine evaluation of patients undergoing surgery for possible parathyroid adenoma. Their usefulness in directing a conservative unilateral operation may result in time and cost savings, as well as reduced surgical exposure. PMID- 8230383 TI - Unilateral parathyroidectomy: the value of the localizing scan. AB - Surgery for correction of primary hyperparathyroidism utilizing a standard bilateral neck exploration has a success rate of approximately 90 to 95%. With the inception of pre-operative localization studies that were 90% accurate in localizing the diseased gland, the concept arose that a unilateral exploration could be as successful as a bilateral exploration. Bilateral exploration of the neck for hyperparathyroidism exposes the patient to a greater potential of morbidity for hypoparathyroidism and recurrent laryngeal nerve injury. It is our feeling based on personal experience that unilateral parathyroidectomy in selective cases can be as successful as the bilateral operation and be more cost effective, saving over $1,100 (U.S.) per case. PMID- 8230384 TI - The role of CT imaging in the evaluation of cervical esophageal foreign bodies. AB - Foreign bodies in the hypopharynx and cervical esophagus, such as chicken and fish bones, occur frequently and usually need radiologic work-up in order to demonstrate the presence of the foreign body and its location. Plain antero posterior and lateral X-ray views of the neck, and if needed, a barium swallow, are the standard. When these studies fail to show the foreign body, unenhanced CT may demonstrate these small calcified esophageal foreign bodies. We evaluated 13 patients with cervical esophageal foreign bodies by CT. It is readily available and rapid and exposes the patient to less radiation than a barium swallow. CT evaluation with soft tissue and bone windows may replace the barium swallow because of its better detection of thin, small, minimally calcified foreign bodies which are often obscured by overlying tissues in the usual X-ray studies. PMID- 8230385 TI - Ultrasound of the neck. AB - In many institutions sonography still is used for evaluation of the thyroid gland and for assessment of sclerotic lesions of the carotid arteries only. The aim of this article is to summarize the potentials and limitations of sonography in the investigation of the extrathyroidal cervical soft tissues. The knowledge of the anatomy of this complex region is mandatory for the investigation of cervical soft tissue lesions. PMID- 8230386 TI - CT manifestations of cervical tuberculous lymphadenitis. AB - Cervical tuberculous lymphadenitis is a commonly encountered disease, especially in Korea. Differentiation from other metastatic lymphadenopathies and benign conditions such as cystic neck masses is important. CT findings of tuberculous lymphadenopathy in the abdomen and thorax are frequently reported in the literature, though there are only a few articles concerning cervical tuberculous lymphadenopathy. The authors retrospectively analyzed CT findings of 42 cases with cervical tuberculous lymphadenitis regarding distribution, contour, enhancing pattern, changes in the adjacent fascial plane, and dermal and subcutaneous manifestations. We concluded that the following are suggestive of tuberculous lymphadenitis: the presence of conglomerated nodal masses with central lucency, thick irregular rim of contrast enhancement and inner nodularity, varying degree of homogenous enhancement in smaller nodes, dermal and subcutaneous manifestations of inflammation such as thickening of the overlying skin, engorgement of the lymphatics and thickening of the adjacent muscles, and a diffusely effaced fascial plane. There were only a few cases of central low densities with calcifications on CT scan. However, common CT patterns of tuberculous adenitis may also be seen in other diseases, such as lymphoma and metastatic lymphadenopathy, and a thorough clinical examination and detection of primary lesions may be more helpful in differentiating these lymphadenopathies. PMID- 8230387 TI - Sport biomechanics 2000. PMID- 8230388 TI - The effects of increased shooting distance in the basketball jump shot. AB - Three-dimensional video techniques (50 Hz) were used to obtain images of basketball jump shots from one of three distances--short range (group 1, n = 5); medium range (group 2, n = 5); long range (group 3, n = 5)--from the basket, as performed by members of the men's quarter-finalist teams at the games of the XVI Universiade in Sheffield in 1991. Fifteen sequences were digitized, beginning 20 frames prior to take-off to 10 frames after release. To facilitate analysis, the sequences were rotated about the ball position in the final frame so that the shot direction was parallel to one of the pre-defined orthogonal axes. Mean (+/- 1 S.D.) ball release speed was found to increase with distance from the basket (group 1 = 3.04 +/- 0.65 m s-1, group 2 = 4.71 +/- 0.74 m s-1, group 3 = 6.24 +/- 0.80 m s-1), while mean release angles were similar for all groups (group 1 = 48.8 +/- 10.1 degrees, group 2 = 47.8 +/- 5.8 degrees, group 3 = 51.9 +/- 5.5 degrees). The increased impulse necessary for the ball to reach the basket at increased shooting distances was derived from both an increase in angular velocity of the elbow joint of the shooting arm and an increased velocity of the centre of mass in the direction of the basket at release. Centre of mass speed at take-off was found to be influenced to a greater extent by the angular velocity of the ankle joint than that of the knee or hip joints.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230389 TI - A three-dimensional cinematographical analysis of the volleyball spike. AB - The objectives of this study were to describe the volleyball spiking actions used by players in top-level competition, and also to examine the interrelationships between upper limb, lower limb and whole body kinematic variables, and post impact ball speed in the spiking technique. Two Photosonics Biomechanics 500 cine cameras operating at a nominal frame rate of 100 Hz were used to film the spiking actions of 10 male senior international volleyball players at the XVI Universiade (1991 World Student Games). Three-dimensional object space co-ordinates of digitized image co-ordinates were obtained using a DLT algorithm and an array of calibration points in the filmed volume. Relationships between lower limb angular kinematics at take-off, centre of mass vertical velocity at take-off and centre of mass vertical displacement (jump height) were examined. Relationships between angular kinematics of the hitting arm and post-impact ball speed were also determined. The mean (+/- S.E.) centre of mass vertical velocity at take-off was 3.59 +/- 0.05 m s-1 and the mean height jumped was 0.62 +/- 0.02 m. As expected, a significant correlation was found between the square of the centre of mass vertical velocity at take-off and jump height (r = 0.78; P < 0.01). No significant correlations were found between lower limb angular kinematics and centre of mass vertical velocity at take-off or jump height. The mean post-impact ball speed was 27.0 +/- 0.9 m s-1, and this was significantly correlated to maximum right humerus angular velocity (r = 0.75; P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230390 TI - A biomechanical analysis of the last stride, touch-down and take-off characteristics of the women's long jump. AB - This study was concerned with the measurement of a selection of performance variables from competitors in the women's long jump final of the World Student Games held in Sheffield, UK in July 1991. Several performances of each of six finalists were recorded on cine-film at 100 Hz. Resulting planar kinematic data were obtained for the last stride, touch-down and take-off. For the analysis, the point of maximum knee flexion was established and this was used to represent the point at which the compression phase had ended. A variety of variables describing the position, velocity and angular changes are presented as descriptive data. In addition, these were used to compute energies on the basis of a whole body model. The data were interpreted on the basis of a technique model of long jumping established from the literature. It was confirmed that take-off velocity was a function of touch-down velocity, and that there was an increase in vertical velocity at the expense of a reduction of horizontal velocity. An attempt was made to identify the mechanisms acting during the touch-down to take-off phase which were responsible for generating vertical velocity. It was concluded that there was evidence for mechanical, biomechanical and muscular mechanisms. The former relates to the generation of vertical velocity by the body riding over the base of support; the second is the elastic re-utilization of energy; and the third is the contribution by concentric muscular contraction. PMID- 8230391 TI - A three-dimensional analysis of javelin throwing technique. AB - This paper reports a three-dimensional kinematic analysis of the best throws from the women's and men's javelin competitions at the 1991 World Student Games in Sheffield. This was undertaken to obtain accurate three-dimensional release parameter values for elite javelin throwers in a top competition, to relate the release parameters to important temporal and kinematic features of the javelin throwing technique and to carry out an assessment of the adequacy of two dimensional estimates of release parameter values in comparison with those from the three-dimensional analysis. Cine-film was taken using two phase-locked Panasonic 1PL cameras running at a frame rate of 100 Hz, and every other frame was digitized. All methodologies were in accord with the guidelines of the British Association of Sports Sciences. The results indicated the possibility of two different throwing techniques, within and between individuals, relating both to the timing of hip joint actions within the kinematic chain and the trade-off between the impulse applied to the javelin and run-up speed. The results also indicated that important release parameter values assessed from the xz-plane alone differed only marginally from those from three dimensions. The extra information obtainable from three-dimensional analysis (such as angle of sideslip, incidence magnitude, azimuth angle, angle of track and yaw rate) along with the poor representation of the time-course of important variables obtained from the xz-plane makes the three-dimensional analysis of this event essential. This is further strengthened by the practical use of three-dimensional analysis in allowing coaches and throwers the opportunity to view the event from any perspective. Recommendations for future research include further three dimensional kinematic analyses with priority being given to assessing ways of estimating wind conditions around the javelin runway and of incorporating the vibrations of the javelin within the analysis. PMID- 8230392 TI - Hand placement techniques in long horse vaulting. AB - In this study, the effects of two different hand placement techniques used by gymnasts to perform Tsukahara and Kasamatsu long horse vaults were examined. Selected linear and angular flight descriptors were calculated to determine whether those gymnasts making initial hand contact on the end of the horse gained additional lift, range or rotation when compared to those gymnasts making the more traditional initial hand contact on top of the horse. Three-dimensional cine film analysis using the Direct Linear Transformation (DLT) was used to obtain data on 17 elite gymnasts competing in the 1991 World Student Games at Sheffield, UK. The gymnasts were divided into two groups according to the techniques used: group E in which the first hand contact was made on the vertical surface of the near end and the second on the top of the horse, and group T in which both hands were placed on top of the horse. The vertical and horizontal motion of each gymnast's mass centre and the somersault rotation during pre-flight (board take off to horse contact) and post-flight (horse take-off to ground landing) were determined. The projections of linear displacements of each gymnast's mass centre onto a vertical plane were determined from the three-dimensional mass centre co ordinates, and somersault angles were calculated using the line joining the midpoints of each gymnast's shoulders and knees. Whole body mass centre linear velocity and somersault angular velocity were determined using quintic splines.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230393 TI - Twisting techniques used by competitive divers. AB - At the 1991 World Student Games, eight reverse 1 1/2 somersault dives with 2 1/2 twists were recorded during the men's finals in the 1 m and 3 m springboard diving competitions using two video cameras. Angles describing body configuration were determined from video data and were input, together with initial orientation angle values and angular momentum components, into a computer simulation model of aerial movement in order to predict body orientation in space. Mean absolute deviations between simulation and video after the completion of one twist were 0.02 rev for somersault, 2.3 degrees for tilt and 0.04 rev for twist. Contributions to the tilt angle after one twist were used as measures of the twisting potential of various techniques and were determined using simulations based on modifications of the video data. Seven of the eight competitors produced the majority of the tilt using aerial techniques which were predominantly asymmetrical movements of the arms and hips, although the mean contribution from contact techniques amounted to one-third of the total tilt. PMID- 8230394 TI - Sports science in the twenty-first century. PMID- 8230395 TI - Comparative kinetic disposition of oxfendazole in sheep and goats before and during infection with Haemonchus contortus and Trichostrongylus colubriformis. AB - The kinetic disposition of [14C]-oxfendazole (OFZ) and its metabolites, fenbendazole (FBZ) and fenbendazole sulphone (FBZ.SO2), in plasma and abomasal fluid were determined in Merino sheep and Angora goats before and during infection with Trichostrongylus colubriformis and Haemonchus contortus. The systemic availability (area under the plasma curve, AUC) of OFZ was significantly lower in goats (13.5 micrograms.h/ml) than in sheep (22.2 micrograms.h/ml) and was reduced with infection in goats (5.6 micrograms.h/ml) and sheep (15.1 micrograms.h/ml). The elimination of plasma [14C] was faster in goats than in sheep. The responses observed for [14C] were a reflection of the behaviour of OFZ. The concentration of OFZ and metabolites in abomasal fluid were similar in both species in the absence or presence of infection. However, as the mean flow rate of abomasal fluid was slower in goats (240 ml/h) than in sheep (488 ml/h), only 7% of the dose passed the pylorus in abomasal fluid of goats compared with 14% in sheep. The presence of gastrointestinal nematodes generally increased abomasal fluid flow rate but neither species nor infection had any effect on the rate or extent of [14C] excretion in urine or faeces. It is suggested that goats possess a faster hepatic metabolism than sheep resulting in more rapid elimination of OFZ. PMID- 8230396 TI - Comparative pharmacokinetic disposition of closantel in sheep and goats. AB - The pharmacokinetic disposition of closantel was examined following intraruminal (i.r.) or intramuscular (i.m.) administration to adult Merino sheep and to adult and 3-month-old, suckling Angora goats. In adult goats the maximum concentration (Cmax) and area under the plasma concentration with time curve (AUC) following 3.75, 7.5 and 15.0 mg closantel/kg given i.r. increased with dose however the time of Cmax (Tmax = 2.6d) in plasma was unaffected by dose rate. The elimination phase (K10) of closantel was monoexponential with a half-life (t1/2) of 4.7d again unaffected by dose rate. Apart from a more rapid absorption phase and earlier Tmax following 3.75 mg closantel/kg i.m., pharmacokinetic behaviour was similar to that following i.r. administration at 3.75 or 7.5 mg/kg. Although absorption rate was more rapid in kids after i.r. administration at 7.5 mg/kg, pharmacokinetic disposition of closantel was otherwise similar to that in adult goats. No closantel was detected in milk of treated doses or in the plasma of their kids. I.R. closantel at 7.5 mg/kg was more slowly absorbed in goats than in sheep but Cmax was similar in both species. However, K10 t1/2 was significantly shorter in goats (4d) than in sheep (14d). Faster elimination resulted in an almost three-fold lowering of AUC in goats and could dramatically reduce the sustained action of closantel in this species compared with sheep. PMID- 8230397 TI - The influence of Ostertagia circumcincta and Trichostrongylus colubriformis infections on the pharmacokinetics of febantel in lambs. AB - Plasma concentrations of febantel and its major metabolites fenbendazole, oxfendazole and oxfendazole sulphone were determined after oral administration of 7.5 mg/kg febantel in lambs before and 28 days after infection with 50,000 L3 larvae of Ostertagia circumcincta or Trichostrongylus colubriformis. The febantel concentrations were always very low and only in a few samples higher than the detection limit. The mean decrease in AUC for the three metabolites for the infected sheep in comparison to the parasite naive sheep was 13.9% +/- 4.1% (mean +/- SEM) and 23.7% +/- 5.3% in the O. circumcincta infected and the T. colubriformis infected lambs respectively. This reduction was only significant for the T. colubriformis infected group. In order to determine a more complete pharmacokinetic profile, febantel was injected intravenously at a dose of 2.5 mg/kg in a further study. PMID- 8230398 TI - Plasma salicylate concentrations in immature dogs following aspirin administration: comparison with adult dogs. AB - Aspirin disposition in immature and adult dogs, assessed by plasma salicylate concentrations following single doses of aspirin given orally (p.o.) and intravenously (i.v.), was compared. Using a cross-over design, four immature (12 16-weeks-old) and eight adult (1-2-years-old) dogs were given a single dose of aspirin at 17.5 mg/kg body weight i.v. and a single dose of buffered aspirin at 35 mg/kg body weight p.o. Blood was collected from the jugular vein for 24 h following each dose. A fluorescence polarization immunoassay was used for determination of salicylate in plasma. Significant differences in aspirin disposition were identified between the two groups. Immature dogs had significantly shorter salicylate half-life, lower mean residence time, and more rapid salicylate clearance than adult dogs. The difference in volume of distribution between the two groups was not significantly different. Immature dogs had lower mean (+/- SD) peak plasma salicylate concentrations (64.5 +/- 2.38 mg/L) than adult dogs (95.9 +/- 12.2 mg/L) following a single oral dose of buffered aspirin at 35 mg/kg body weight. Predicted plasma salicylate concentration-time curves were constructed for various aspirin dosage regimens. This analysis showed that the previously recommended buffered aspirin dose for adult dogs of 25 mg/kg body weight p.o. every 8 h would be ineffective in maintaining plasma salicylate concentrations > 50 mg/L in immature dogs. PMID- 8230399 TI - The disposition of suxibuzone in the horse. AB - A high performance liquid chromatographic method is described to determine the anti-inflammatory drug suxibuzone (SXB) and its major metabolites phenylbutazone (PBZ) and oxyphenbutazone (OPBZ) in equine plasma and urine. When suxibuzone (6 mg/kg) was administered intravenously (i.v.) or orally (p.o.) no parent drug was detected in plasma or in urine. The disposition of the metabolite PBZ (i.v.) could be described by a 2 compartment model with a beta half-life varying from 7.40 to 8.35 h. Due to severe side effects the use of i.v. suxibuzone should not be encouraged in the horse. PBZ and OPBZ were detected in plasma and urine after p.o. SXB administration. Peak plasma PBZ concentrations (8.8 +/- 3.0 micrograms/ml) occurred 6 h after oral dosing and the terminal exponential constant was 0.11 +/- 0.01 h-1. Phenylbutazone and oxyphenbutazone were detectable in urine (> 1 microgram/ml) for at least 36 h, after p.o. administration. SXB was not hydrolyzed in vitro by horse plasma. Equine liver homogenates however appeared to have a very high capacity for hydrolysing SXB, indicating that first-pass effect could be responsible for the rapid disappearance of this NSAID in the horse. PMID- 8230400 TI - Pharmacokinetics of diminazene in female Boran (Bos indicus) cattle. AB - The disposition kinetics and bioavailability of diminazene in five healthy heifers were determined after single intravenous (i.v.) and intramuscular (i.m.) administration of the drug in sequence with a wash-out period between administrations of 6 weeks. Intact diminazene in plasma, whole blood and urine samples was analysed using high-performance liquid chromatography. Nonlinear regression analysis of the i.v. and i.m. data indicated that, for either route, the plasma disappearance curves of diminazene were best described by triexponential equations. The i.v. bolus was followed by rapid and biphasic distribution with half-life values of 0.04 h and 0.58 h, Vd(ss) was 1.91 +/- 0.42 l/kg, elimination half-life was 31.7 h while Cl averaged 1.74 +/- 0.40 ml/min/kg. Within 30 min of the i.v. dose, the erythrocyte/plasma partition ratio of diminazene was 0.30 +/- 0.15. Diminazene was rapidly absorbed following i.m. administration; t1/2ka was 0.60 h. Cmax, 4.68 +/- 1.12 micrograms/ml, was attained in 10-15 min and systemic availability was 102.42 +/- 7.25%. The half life of the terminal disappearance phase was 145.48 h. About 8.26% of the i.m. dose was excreted intact in the urine within the first 24 h of treatment. In vitro, diminazene was bound to bovine plasma albumin to the extent of 38.01 91.10%. PMID- 8230401 TI - Toxicologic effects of ribavirin in cats. AB - Ribavirin, a broad-spectrum antiviral agent active in vitro against a number of RNA and DNA viruses, has been associated with moderate toxicity in laboratory animals and humans. Clinically, ribavirin has been used effectively in persons primarily to treat life-threatening viral diseases such as acute haemorrhagic fever or viral pneumonia of infants. In order to evaluate the feasibility of using this antiviral agent in cats, the effects of oral (p.o.), intramuscular (i.m.) and intravenous (i.v.) doses of ribavirin in 27 9-month-old specific pathogen-free cats were evaluated by haematology, clinical chemistries, bone marrow biopsies and histopathology. Ribavirin was administered once daily for 10 consecutive days at a dose of either 11, 22, or 44 mg/kg after which all cats were euthanatized and necropsied. Most cats receiving 22 or 44 mg of ribavirin/kg became anorectic and suffered some degree of weight loss (0.2 to 0.6 kg), and about one-third of the cats developed diarrhoea and/or mucous membrane pallor. Icterus or haemorrhage was not observed. The most profound and consistent haematologic change, particularly among the moderate and high dosage groups regardless of route of administration, was a significant and severe thrombocytopenia (range, 33-78% reduction in mean platelet counts vs. baseline). Other changes, particularly reductions in total WBC and neutrophils and reductions in RBC and PCV, tended to occur at lower ribavirin dosages, but generally they were not statistically significant. Cats given 44 mg of ribavirin/kg i.v. showed significant decreases in leukocyte variables, including total WBC (P = 0.016), neutrophils (P = 0.026) and lymphocytes (P = 0.047). Mild to-moderate increases in serum alanine aminotransferase and alkaline phosphatase activities occurred at doses of 22 and 44 mg/kg. Evaluation of bone marrow biopsies before and after treatment revealed that cats given 11 mg of ribavirin/kg had mild megakaryocytic (MK) hypoplasia, whereas cats receiving 22 or 44 mg/kg had progressively severe degrees of MK hypoplasia and dysplasia, asynchronous MK maturation, and increased myeloid:erythroid ratio. Pathologic changes in ribavirin-treated cats generally were mild and included primarily enteritis (seven cats) and hepatocellular vacuolation and/or centrilobular necrosis (seven cats). Results of this study in cats indicated that daily administration of ribavirin at a dose range of 11 to 44 mg/kg induced a dose related toxic effect on bone marrow, primarily on megakaryocytes and erythroid precursors, and at the higher dosages is suppressed numbers of circulating leukocytes. PMID- 8230402 TI - A study of the disposition of procaine penicillin G in feedlot steers following intramuscular and subcutaneous injection. AB - The disposition of an aqueous suspension of procaine penicillin G (300,000 U/mL) was studied in feedlot steers. Four groups of three steers were used. Steers in groups 1 and 2 received procaine penicillin G once daily for 5 days intramuscularly (i.m.) at a dose of 24,000 U/kg (group 1) or of 66,000 U/kg (group 2). The injection on the last day was administered in the gluteal muscle. Steers in group 3 (i.m. neck injection) and group 4 [subcutaneous (s.c.) injection] each received a single dose of procaine penicillin G at a dose of 66,000 U/kg. From every animal, after the last injection in groups 1 and 2 and following the single injection in groups 3 and 4, a series of blood samples was taken at fixed time intervals. The plasma from these samples was analysed for penicillin G by a high performance liquid chromatography (HPLC) assay in order to determine the disposition of penicillin. The maximum plasma concentration (Cmax) and the area under the curve (AUC) were significantly different between groups 1 and 2, but we found no difference in the disappearance rate constant between these two groups. Group 4 single s.c. injections produced a lower mean Cmax (1.85 +/- 0.27 microgram/mL) than the mean Cmax (4.24 +/- 1.08 micrograms/mL) produced in group 3 by i.m. injections into the neck muscle or the mean Cmax (2.63 +/- 0.27 microgram/mL) produced in group 2 by i.m. injections into the gluteal muscle. However the mean Cmax produced by i.m. injections into the neck muscles (group 3) was higher than the mean Cmax produced by i.m. injections into the gluteal muscle (group 2). Additionally, the disappearance t1/2 was longer (18.08 h) in group 4 following the s.c. injection and shorter (8.85 h) in group 3 following the i.m. neck injection, than the t1/2 following administration of the same dose i.m. into the gluteal muscle (15.96 h) in group 2. In this study, when procaine penicillin G was injected into the gluteal muscle, doses of 66,000 U/kg were necessary to produce plasma concentrations that were above a minimum inhibitory concentration (MIC) for penicillin G of 1.0 microgram/mL as compared to doses of 24,000 U/kg. PMID- 8230403 TI - Regulation of uterine estrogen receptors (ER) by beta-adrenergic stimulation in immature rats. AB - The effects of a 3-day intramuscular (i.m.) administration of clenbuterol (25 micrograms/Kg), propranolol (12 mg/kg), clenbuterol (25 micrograms/kg) plus propranolol (12 mg/Kg) and estradiol (0.5 microgram) upon the female reproductive system were investigated in immature Sprague-Dawley rats. Clenbuterol and estradiol treatments induced a significant increase in uterus weight and in relative uterus weight, whereas in the groups treated with propranolol and clenbuterol plus propranolol no differences were detected versus controls. The uterine estrogen receptor levels were significantly increased by clenbuterol administration. In the rats dosed with propranolol and clenbuterol plus propranolol, no modifications occurred in estrogen receptor concentrations when compared with control values. Uterine progesterone receptors were never significantly affected by any of the considered treatments. Data obtained indicate that clenbuterol treatment induces an increase in uterus weight and in estrogen receptor levels and that these effects are regulated by acute beta adrenergic stimulation, as the contemporaneous administration of high doses of a beta-blocker inhibit such effects. PMID- 8230404 TI - The effects of halothane and nitrous oxide on the pharmacokinetics of propofol in dogs. AB - The pharmacokinetics of propofol, 6.5 mg/kg, administered as a bolus dose intravenously (i.v.) were studied in six dogs (group 1). The effect of maintaining anaesthesia with halothane and nitrous oxide in oxygen on propofol pharmacokinetics was also investigated in six dogs undergoing routine anaesthesia (group 2). Induction of anaesthesia was rapid in all animals. Post-induction apnoea was a feature in three of the 12 dogs. The blood propofol concentration time profile was best described by a bi-exponential decline in two dogs in group 1 and in 3 dogs in group 2, and by a tri-exponential decline in four dogs in group 1 and 3 dogs in group 2. The elimination half-life was long in both groups (90.9 min and 75.2 min, respectively), the volume of distribution at steady state large (4889 and 4863 ml/kg) and the clearance rapid (58.6 and 56.3 ml/kg.min). There were no significant differences between the groups, thus indicating that maintenance of anaesthesia with halothane and nitrous oxide had no effect on the pharmacokinetics of propofol in the dog. PMID- 8230405 TI - Sulphadimidine metabolism in vitro: I. Sex differences in acetylation and hydroxylation in cultured rat hepatocytes. AB - The hydroxylation and acetylation of 0.5 mM sulphadimidine (SDD) was studied in primary cultures of hepatocytes from male and female rats, and from castrated male and sham operated male rats. In addition, SDD metabolism was investigated in hepatocytes from castrated male rats treated with testosterone, prior to liver cell isolation. In male rat hepatocytes a significantly higher hydroxylation activity was observed than in hepatocytes from female and castrated male rats. Acetylation activity was higher in females. Testosterone induced hydroxylation but did not affect acetylation. These results correlate well with data from previous in vivo studies, showing the relevance of this in vitro model. PMID- 8230406 TI - Influence of yohimbine and tolazoline on the cardiovascular, respiratory, and sedative effects of xylazine in the horse. AB - To determine the effects of yohimbine and tolazoline on the cardiovascular, respiratory and sedative effects of xylazine, four horses were sedated with xylazine and treated with either yohimbine, tolazoline or saline. Xylazine was administered as an intravenous (i.v.) bolus (1.0 mg/kg) followed by a continuous infusion at the rate of 12 micrograms/kg/min. Heart rate, respiratory rate, mean arterial pressure, arterial blood gases, and the chin-to-floor distance were recorded throughout the experiment. After 60 min, either yohimbine or tolazoline was administered i.v. in incremental doses until reversal of sedation (defined as the return of the chin-to-floor distance to baseline values) was achieved. A control group in which a saline bolus was administered instead of an antagonist drug was included for comparison. The average dose of yohimbine administered was 0.12 +/- 0.02 (SEM) mg/kg. While the average dose of tolazoline was 7.5 +/- 1.1 mg/kg. Both tolazoline and yohimbine antagonized the ventricular bradycardia and A-V conduction disturbances observed with xylazine administration. No change in mean arterial pressure was observed with xylazine or yohimbine administration, but tolazoline caused persistent mild systemic hypertension. There were no clinically significant changes in respiratory rate or arterial blood gas values with administration of either xylazine, yohimbine or tolazoline. The chin-to floor distance decreased significantly with xylazine administration and increased significantly with administration of either yohimbine or tolazoline. In conclusion, both yohimbine and tolazoline successfully antagonized the cardiovascular and CNS depression associated with xylazine administration. PMID- 8230407 TI - Disposition, bioavailability and clinical efficacy of orally administered acepromazine in the horse. AB - The pharmacokinetics and pharmacological efficacy of orally (p.o.) administered acepromazine were studied and compared with the intravenous (i.v.) route of administration in a cross-over study using six horses. The oral kinetics of acepromazine can be described by a two-compartment open model with first-order absorption. The drug was rapidly absorbed after p.o. administration with a half life of 0.84 h, tmax of 0.4 h and Cmax of 59 ng/ml. The elimination was slower after p.o. administration (half-life 6.04 h) than after i.v. injection (half-life 2.6 h). The bioavailability of the orally administered drug formulation was 55.1%. After p.o. administration of 0.5 mg/kg acepromazine, the parameters of the sedative effect were similar to those obtained after i.v. injection of 0.1 mg/kg. The effect of the drug on blood cell count and haemoglobin content was similar after both p.o. administration and injection, while the effects on the parameters of penile prolapse and on the mean arterial blood pressure were less pronounced after p.o. administration than after injection. After p.o. administration, no significant effects on haematocrit-level as well as on the heart and respiratory rates were observed, while these parameters were significantly affected after injection. It is concluded that the high initial plasma level of the drug after i.v. injection may play a role in producing adverse effects of acepromazine. PMID- 8230408 TI - Duration of analgesia induced by epidurally administered morphine and medetomidine in dogs. PMID- 8230409 TI - Enantioselective pharmacokinetics of ketoprofen in horses. PMID- 8230410 TI - Plasma concentrations of fenbendazole and its metabolites in poultry after a single oral administration. PMID- 8230411 TI - ATCvet--a classification system for veterinary medicinal products. PMID- 8230412 TI - Picornavirus nonstructural proteins: emerging roles in virus replication and inhibition of host cell functions. PMID- 8230413 TI - Induction of gene expression by exon 2 of the major E1A proteins of adenovirus type 5. AB - We have constructed an adenovirus type 5 (Ad5) E1A mutant, dl1119/520, that produces essentially only exon 2 of the major E1A proteins. In infected primary baby rat kidney cells, this mutant induced expression of the E1B 55-kDa protein, and in infected human KB cells, it induced expression of this protein, the E2A 72 kDa protein, and hexon. In KB cells, this mutant grew substantially better than Ad5 dl312, which lacks E1A, and as well as Ad5 dl520, an E1A mutant producing only the 243-residue protein. These results suggest that exon 2 of E1A proteins on its own was able to activate gene expression. We also constructed mutants of dl1119/520, containing small deletions in regions of exon 2 that others found to be associated with effects on the properties of E1A transformants. None of these deletions destroyed gene activation completely, indicating that there may be some redundancy among sequences in exon 2 for inducing gene expression. The two deletions that decreased induction the most, residues 224 to 238 and 255 to 270, were in regions reported to be associated with the expression of a metalloprotease and with enhanced transformation, suggesting that exon 2 may regulate expression of genes governing cell growth. It is remarkable that all sections of E1A proteins, exon 1, the unique region, and exon 2, have now been found to affect gene expression. PMID- 8230414 TI - Efficient self-assembly of human papillomavirus type 16 L1 and L1-L2 into virus like particles. AB - The L1 genes of two human papillomavirus type 16 (HPV16) isolates derived from condylomata acuminata were used to express the L1 major capsid protein in insect cells via recombinant baculoviruses. Both L1 major capsid proteins self-assembled into virus-like particles (VLP) with high efficiency and could be purified in preparative amounts on density gradients. The yield of VLP was 3 orders of magnitude higher than what has been obtained previously, using L1 derived from the prototype HPV16. DNA sequence comparison identified a single nonconserved amino acid change to be responsible for the inefficient self-assembly of the prototype L1. VLP were also obtained by expressing L1 of HPV6, HPV11, and cottontail rabbit papillomavirus, indicating that L1 from a variety of papillomaviruses has the intrinsic capacity to self-assemble into VLP. Coexpression of HPV16 L1 plus L2 by using a baculovirus double-expression vector also resulted in efficient self-assembly of VLP, and the average particle yield increased about fourfold in comparison to when L1 only was expressed. Coimmunoprecipitation of L1 and L2 and cosedimentation of the two proteins in a sucrose gradient demonstrated that L2 was incorporated into the particles. The ability to generate preparative amounts of HPV16 L1 and L1-L2 VLP may have implications for the development of a serological assay to detect anti-HPV16 virion immune responses to conformational epitopes and for immunoprophylaxis against HPV16 infection. PMID- 8230415 TI - Increased spacing between Sp1 and TATAA renders human immunodeficiency virus type 1 replication defective: implication for Tat function. AB - Expression of the human immunodeficiency virus type 1 (HIV-1) is strongly activated by Tat. The proper action of Tat requires three elements: TATAA, TAR, and upstream motifs in the HIV-1 long terminal repeat. We show here that the correct spatial arrangement among Tat, Sp1, and TATAA crucially influences HIV expression. Under conditions in which basal promoter activity is unperturbed, distancing Sp1 from TATAA markedly affected Tat trans activation. An increase in the Sp1-TATAA distance from 18 to 101 nucleotides (depending on the inserted sequence) rendered HIV-1 either partially or wholly replication defective. This critical dependence on spacing suggests that Tat-, Sp1-, and TATAA-binding factors must correctly contact each other for optimal expression and replication of HIV-1. PMID- 8230416 TI - Renaturation of complementary DNA strands by herpes simplex virus type 1 ICP8. AB - ICP8, the major single-stranded DNA-binding protein of herpes simplex virus type 1, promotes renaturation of complementary single strands of DNA. This reaction is ATP independent but requires Mg2+. The activity is maximal at pH 7.6 and 80 mM NaCl. The major product of the reaction is double-stranded DNA, and no evidence of large DNA networks is seen. The reaction occurs at subsaturating concentrations of ICP8 but reaches maximal levels with saturating concentrations of ICP8. Finally, the renaturation reaction is second order with respect to DNA concentration. The ability of ICP8 to promote the renaturation of complementary single strands suggests a role for ICP8 in the high level of recombination seen in cells infected with herpes simplex virus type 1. PMID- 8230417 TI - Multiple double-stranded RNA segments are associated with virus particles infecting Trichomonas vaginalis. AB - Previous studies demonstrated that some isolates of the sexually transmitted protozoan Trichomonas vaginalis are infected with a nonsegmented, double-stranded RNA (dsRNA) virus. A reexamination of the total dsRNA extracted from several virus-harboring isolates indicated the presence of at least three dsRNAs with sizes ranging from 4.8 to 4.3 kbp. The double-stranded nature of each of the three segments was determined by hybridization experiments using riboprobes of opposite polarities obtained from cDNA generated to each of the segments. All three segments were present in agar clones originating from single organisms of T. vaginalis isolates, suggesting that the three segments were not the result of a mixed population of trichomonads harboring different sizes of dsRNA. The three segments were associated with CsCl-purified virus particles, as evidenced by electron microscopy, and RNAse treatment of the preparation containing virus particles did not destroy the dsRNAs. Finally, the individual dsRNA segments were purified for use as probes to determine whether the three dsRNAs shared any sequence homology. Each end-labeled dsRNA segment did not cross-hybridize to any of the other two segments, a finding consistent with the hybridization of labeled cDNAs to only the segments from which they were derived. These results show that the coding capacity of the dsRNA virus may be at least three times greater than that estimated earlier and illustrates further the complexity of this virus parasite interrelationship. PMID- 8230418 TI - Effects of the tat and nef gene products of human immunodeficiency virus type 1 (HIV-1) on transcription controlled by the HIV-1 long terminal repeat and on cell growth in macrophages. AB - The RAW264 murine macrophage cell line was used as a model to examine the role of the tat and nef gene products in the transcription regulation of the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) in macrophages. Contrary to claims that the activity of the HIV-1 LTR responds poorly in rodent cells to trans activation by the viral tat gene product, cotransfection of RAW264 cells with a tat expression plasmid in transient transfection assays caused a > 20-fold increase in reporter gene expression that was inhibited by mutations in the TAR region. RAW264 cells stably transfected with the tat plasmid displayed similarly elevated HIV-1 LTR-driven reporter gene activity. By contrast to previous reports indicating a negative role for nef in HIV transcription, cotransfection of RAW264 cells with a nef expression plasmid trans activated the HIV-1 LTR driving either a chloramphenicol acetyltransferase or a luciferase reporter gene. The action of nef was specific to the LTR, as expression of nef had no effect on the activity of the simian virus 40, c-fms, urokinase plasminogen activator, or type 5 acid phosphatase promoter. trans-activating activity was also manifested by a frameshift mutant expressing only the first 35 amino acids of the protein. The effects of nef were multiplicative with those of tat gene product and occurred even in the presence of bacterial lipopolysaccharide, which itself activated LTR-directed transcription. Examination of the effects of selected mutations in the LTR revealed that neither the kappa B sites in the direct repeat enhancer nor the TAR region was required as a cis-acting element in nef action. The action of nef was not species restricted; it was able to trans activate in the human monocyte-like cell line Mono Mac 6. The presence of a nef expression cassette in a neomycin phosphotransferase gene expression plasmid greatly reduced the number of G418 resistant colonies generated in stable transfection of RAW264 cells, and many of the colonies that were formed exhibited very slow growth. The frameshift mutant was also active in reducing colony generation. Given the absence of any effect of the frameshift mutation on nef function, its actions on macrophage growth and HIV transcription are discussed in terms of the role of the N-terminal 30 amino acids and of stable secondary structures in the mRNA. PMID- 8230419 TI - The RNAs of hepatitis delta virus are copied by RNA polymerase II in nuclear homogenates. AB - Human hepatitis delta virus has a single-stranded circular RNA genome that replicates by RNA-directed RNA synthesis. The virus encodes only a single protein, the delta antigen, which both is small (22 kDa) and lacks sequence homology to known RNA polymerases, suggesting that the virus employs a cellular polymerase for replication. Consistent with this suggestion, we have used homogenized nuclei from a human hepatoma cell line, HepG2, to demonstrate RNA directed RNA synthesis from both genomic hepatitis delta virus RNA and its complement, the antigenomic RNA. RNA polymerase II was responsible for this transcription because the reaction was inhibited both by low doses of alpha amanitin and by a monoclonal antibody specific for polymerase II. In addition, it was found that the majority of the RNA products were processed, presumably by self-cleavage and self-ligation, to produce covalently closed circular molecules. PMID- 8230420 TI - Patterns of frog virus 3 DNA methylation and DNA methyltransferase activity in nuclei of infected cells. AB - The iridovirus frog virus 3 (FV3) can replicate in culture in fat head minnow (FHM) fish cells or in BHK-21 hamster cells. Viral DNA replication commences about 3 h after infection of FHM cells with FV3. Between 3 and 6 h postinfection (p.i.), a portion of the intranuclear FV3 DNA is partly unmethylated. At later times, p.i., all of the viral DNA in the nuclear and cytoplasmic compartments is methylated at the 5'-CCGG-3' sequences. Cytoplasmic FV3 DNA has not been found unmethylated. We have cloned viral DNA fragments from methylated virion DNA. By using the genomic sequencing technique, it has been demonstrated for segments of the FV3 DNA replicated both in FHM fish and BHK21 hamster cells that in a stretch encompassing a total of 350 bp, all of the analyzed 5'-CG-3' dinucleotides are methylated. The modified nucleotide 5-methyldeoxycytidine is present exclusively in the 5'-CG-3' dinucleotide combination. In the cloned FV3 DNA fragment p21A, an open reading frame has been located. The 5' region of this presumptive viral gene is also methylated in all 5'-CG-3' positions. DNA methyltransferase activity has been detected in the nuclei of FV3-infected FHM cells at 4, 11, and 20 h p.i. In the cytoplasmic fraction, comparable activity has not been observed. These data are consistent with the interpretation that FV3 DNA is newly synthesized and de novo methylated in the nuclei of infected FHM cells and subsequently exported into the cytoplasm for viral assembly. PMID- 8230421 TI - Eleven loci encoding trans-acting factors are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA replication. AB - Recently we described the use of human cytomegalovirus (HCMV) cosmid clones in a cotransfection assay of HCMV oriLyt replication (G. S. Pari, M. A. Kacica, and D. G. Anders, J. Virol. 67:2575-2582, 1993). We have now used this assay to identify 11 distinct required loci encoding trans-acting factors sufficient for transient complementation of oriLyt-dependent DNA replication. This set includes all of the virus genes essential to initiate and perform DNA synthesis together with the virus genes required to express these replication functions from their native promoters. Six of the identified loci span open reading frames (ORFs) that encode homologs or probable homologs of herpes simplex virus type 1 replication genes, consistent with predictions based on sequence similarities and biochemical properties. These include the DNA polymerase UL54 and polymerase-associated protein UL44, the single-stranded-DNA-binding protein UL57, and proposed subunits of a helicase-primase complex, UL70, UL105, and UL101-102. Frameshift mutations in any one of these essential ORFs abrogated complementation of DNA replication. Three required loci, UL36-38, IRS1 (or TRS1), and IE1/IE2, encode known regulatory proteins. The remaining two loci span ORFs UL84 and UL112-113 and encode early temporal class nucleus-associated proteins of unknown function. Neither of these genes have been implicated previously in DNA replication or in regulating gene expression, nor have counterparts in herpes simplex virus type 1 or Epstein-Barr virus been described. The results presented here will facilitate investigation of the mechanisms and regulation of HCMV lytic-phase DNA replication. PMID- 8230422 TI - Superinfection-induced apoptosis and its correlation with the reduction of viral progeny in cells persistently infected with Hz-1 baculovirus. AB - Differential induction of necrosis or apoptosis was found upon challenge of cells of the insect Spodoptera frugiperda productively or persistently infected with Hz 1 baculovirus, respectively. Unlike parental SF9 cells, which were essentially all killed by virally induced necrosis, persistently infected cells underwent a process of massive cell death by apoptosis; cells which were not killed by apoptosis then reestablished a cell monolayer. Upon viral challenge, the yield of viral progeny was reduced greatly in persistently virus-infected cells but not in parental cells. Immunolabelling of individual cells revealed that upon viral challenge, production of viral progeny was detectable only in necrotic cells and not in apoptotic cells. These results indicated that induction of apoptosis greatly reduces the yield of viral progeny in cells persistently infected with Hz 1 baculovirus. This is the first report of apoptosis induction in persistently infected cells upon viral superinfection. PMID- 8230423 TI - Internal cleavage and trans-proteolytic activities of the VPg-proteinase (NIa) of tobacco etch potyvirus in vivo. AB - The NIa protein of plant potyviruses is a bifunctional protein containing an N terminal VPg domain and a C-terminal proteinase region. The majority of tobacco etch potyvirus (TEV) NIa molecules are localized to the nucleus of infected cells, although a proportion of NIa is attached covalently as VPg to viral RNA in the cytoplasm. A suboptimal cleavage site that is recognized by the NIa proteinase is located between the two domains. This site was found to be utilized in the VPg-associated, but not the nuclear, pool of NIa. A mutation converting Glu-189 to Leu at the P1 position of the processing site inhibited internal cleavage. Introduction of this mutation into TEV-GUS, an engineered variant of TEV that expresses a reporter protein (beta-glucuronidase [GUS]) fused to the N terminus of the helper component-proteinase (HC-Pro), rendered the virus replication defective in tobacco protoplasts. Site-specific reversion of the mutant internal processing site to the wild-type sequence restored virus viability. In addition, the trans-processing activity of NIa proteinase was tested in vivo after introduction of an artificial cleavage site between the GUS and HC-Pro sequences in the cytoplasmic GUS/HC-Pro polyprotein encoded by TEV GUS. The novel site was recognized and processed in plants infected by the engineered virus, indicating the presence of excess NIa processing capacity in the cytoplasm. The potential roles of internal NIa processing in TEV-infected cells are discussed. PMID- 8230424 TI - Human T-cell leukemia virus type 1 Tax activates transcription of the human fra-1 gene through multiple cis elements responsive to transmembrane signals. AB - We have shown that Tax1 of human T-cell leukemia virus type 1 stimulates the expression of several cellular immediate-early genes (M. Fujii, T. Niki, T. Mori, T. Matsuda, M. Matsui, N. Nomura, and M. Seiki, Oncogene 6:1023-1029, 1991). In this study, the 5'-flanking region of the human fra-1 gene, which is a Tax1 inducible fos-related gene, was isolated and Tax1 or serum-responsive cis elements were analyzed to obtain further insight into the mechanism of Tax1 action. The 62-bp sequence starting 46 nucleotides upstream from the translation initiation site showed 71% homology with the sequence surrounding the TATA box of the c-fos promoter. Regulatory motifs identified in the c-fos promoter, such as an Ets-binding site, E boxes, a CArG box, c-fos AP-1 sites, and two retinoblastoma control elements, were also found upstream of the c-fos homology region. A 502-bp fragment containing these motifs mediated transcriptional activation by Tax1 or by serum in a transient transfection assay. Three independent Tax1-responsive regions (TRRs) were identified, and mutations in each revealed that one of the retinoblastoma control elements in TRR1 and the c-fos AP 1 sites in TRR2 and TRR3 were essential for the activation. Although TRR2 contains a CArG box-like sequence, it was a weak binding site for p67SRF, if it bound at all, and was not required for activation. All three TRRs could also mediate the signals stimulated by serum. Thus, Tax1 appears to activate fra-1 gene expression by means of a part of the cellular machinery similar to that which mediates growth signals. PMID- 8230425 TI - T-cell activation influences initial DNA synthesis of simian immunodeficiency virus in resting T lymphocytes from macaques. AB - The relationship between T-cell activation and early events in the replication cycle of simian immunodeficiency virus (SIV) was analyzed in resting T lymphocytes from macaques. We used the polymerase chain reaction to detect an early product of reverse transcription (R/U5) and almost complete viral DNA (long terminal repeat/gag). We found that SIV can enter resting T lymphocytes and initiate replication but that the reverse transcription process is not efficient and proceeds slowly in resting cells. Cross-linking the CD3/T-cell receptor complex with monoclonal antibodies, unlike cross-linking either the CD28 or CD2 accessory receptor and like phorbol myristate acetate, induced a rapid increase in viral R/U5 DNA detected within 3 to 6 h postinfection. Anti-CD3 or phorbol myristate acetate induced replication of full-length viral DNA within 6 to 9 h postinfection, but full-length SIV DNA was not detectable at earlier time points. We then compared various inhibitors of T-cell activation for their effects on viral initiation and complete replication. Cyclosporin A, an inhibitor of a distal step in T-cell activation, blocked anti-CD3-induced T-cell proliferation and completion of SIV DNA replication but had no effect on induced increases in SIV R/U5 DNA. By contrast, initial SIV DNA synthesis was partially blocked by inhibitors of very early steps in T-cell activation, including herbimycin A, an inhibitor of protein tyrosine kinases, and by two different inhibitors of protein kinase C, H7 and staurosporine. Since resting T cells do not efficiently complete SIV DNA synthesis and cyclosporin A can block the formation of complete viral DNA induced in activated T cells, a cellular factor(s) present in activated T cells appears to be required for the formation of full-length SIV DNA. PMID- 8230426 TI - Aleutian mink disease parvovirus infection of mink macrophages and human macrophage cell line U937: demonstration of antibody-dependent enhancement of infection. AB - Aleutian mink disease parvovirus (ADV) infects macrophages in adult mink. The virulent ADV-Utah I strain, but not the cell culture-adapted ADV-G strain, infects mink peritoneal macrophage cultures and the human macrophage cell line U937 in vitro. However, preincubation of ADV-G with ADV-infected mink serum enhanced its infectivity for U937 cells. the enhancing activity was present in the protein A-binding immunoglobulin G fraction in the serum, but F(ab')2 fragments failed to enhance the infection. On the other hand, the same sera inhibited ADV-G infection of Crandell feline kidney (CRFK) cells. Although U937 cells were not fully permissive for antibody-enhanced ADV-G infection, ADV mRNA expression, genome amplification, and protein expression were identical to those found previously for ADV-Utah I infection of U937 cells. Preincubation of ADV Utah I with soluble protein A partly inhibited the infection of U937 cells but did not affect infection of CRFK cells. In mink peritoneal macrophages, preincubation with the infected mink serum did not make ADV-G infectious. However, the infectivity for mink macrophages of antibody-free ADV-Utah I prepared from the lungs of infected newborn mink kits was enhanced by ADV infected mink serum. Moreover, protein A partly blocked ADV-Utah I infection of mink macrophage cultures. These results suggested that ADV-Utah I enters mink macrophages and U937 cells via an Fc receptor-mediated mechanism. This mechanism, antibody-dependent enhancement, may also contribute to ADV infection in vivo. Furthermore, since ADV infection in mink is characterized by overproduction of anti-ADV immunoglobulins, antibody-dependent enhancement may play a critical role in the establishment of persistent infection with ADV in vivo. PMID- 8230427 TI - Induction of reactivation of herpes simplex virus in murine sensory ganglia in vivo by cadmium. AB - Herpes simplex viruses maintained in a latent state in sensory neurons in mice do not reactivate spontaneously, and therefore the factors or procedures which cause the virus to reactivate serve as a clue to the mechanisms by which the virus is maintained in a latent state. We report that cadmium sulfate induces latent virus to reactivate in 75 to 100% of mice tested. The following specific findings are reported. (i) The highest frequency of induction was observed after two to four daily administrations of 100 micrograms of cadmium sulfate. (ii) Zinc, copper, manganese, or nickel sulfate administered in equimolar amounts under the same regimen did not induce viral reactivation; however, zinc sulfate in molar ratios 25-fold greater than those of cadmium induced viral replication in 2 of 16 ganglia tested. (iii) Administration of zinc, nickel, or manganese prior to the cadmium sulfate reduced the incidence of ganglia containing infectious virus. (iv) Administration of cadmium daily during the first week after infection and at 2-day intervals to 13 days after infection resulted in the recovery from ganglia of infectious virus in titers 10- to 100-fold higher than those obtained from animals given saline. Moreover, infectious virus was recovered as late as 11 days after infection compared with 6 days in mice administered saline. (v) Administration of cadmium immediately after infection or repeatedly after establishment of latency did not exhaust the latent virus harbored by sensory neurons, inasmuch as the fraction of ganglia of mice administered cadmium and yielding infectious virus was similar to that observed in mice treated with saline. We conclude that induction of cadmium tolerance precludes reactivation of latent virus. If the induction of metallothionein genes was the sole factor required to cause reactivation of latent virus, it would have been expected that all metals which induce metallothioneins would also induce reactivation, which was not observed. The results therefore raise the possibility that in addition to inducing the metallothionein genes, cadmium inactivates the factors which maintain the virus in latent state. PMID- 8230428 TI - Dysregulated surface gene expression from disrupted hepatitis B virus genomes. AB - During chronic infection by hepatitis B virus, the viral genome frequently integrates into the host chromosome, causing gross disruption and rearrangement of the viral DNA. We have obtained data showing that viral genomic disruptions which delete the enhancers from the transcribed region of the viral surface gene can lead to dysregulation of surface gene expression at the transcriptional level. Specifically, in cells transfected with such disrupted genomes, there is a decreased amount of transcripts coding for the major form of the surface protein but little change in the amount of transcripts coding for the large surface protein. In these cells, secretion of the surface proteins is blocked in the endoplasmic reticulum-Golgi intermediate compartment, consistent with previous work from other groups showing that relative overexpression of the large surface protein can block secretion of all forms of the surface protein. Our findings suggest that viral genomic rearrangements during integration may be a contributing factor in the pathogenesis of ground-glass hepatocytes, which contain large amounts of intracellular surface proteins as a result of a block in secretion and are frequently seen in the livers of patients with chronic hepatitis B. PMID- 8230429 TI - Characterization of mouse hepatitis virus-specific cytotoxic T cells derived from the central nervous system of mice infected with the JHM strain. AB - The cytotoxic T lymphocyte (CTL) activity of spleen cells from BALB/c (H-2d) mice immunized with the neurotropic JHM strain of mouse hepatitis virus (JHMV) was stimulated in vitro for 7 days. CTL were tested for recognition of target cells infected with either JHMV or vaccinia virus recombinants expressing the four virus structural proteins. Only target cells infected with either JHMV or the vaccinia virus recombinant expressing the JHMV nucleocapsid protein were recognized. Cytotoxic T cell lines were established by limiting dilution from the brains of mice undergoing acute demyelinating encephalomyelitis after infection with JHMV. Twenty of the 22 lines recognized JHMV-infected but not uninfected syngeneic target cells, indicating that they are specific for JHMV. All T-cell lines except one were CD8+. The specificity of the CTL lines was examined by using target cells infected with vaccinia virus recombinants expressing the JHMV nucleocapsid, spike, membrane, and hemagglutinin-esterase structural proteins. Seventeen lines recognized target cells expressing the nucleocapsid protein. Three of the JHMV-specific T-cell lines were unable to recognize target cells expressing any of the JHMV structural proteins, indicating that they are specific for an epitope of a nonstructural protein(s) of JHMV. These data indicate that the nucleocapsid protein induces an immunodominant CTL response. However, no CTL activity specific for the nucleocapsid protein could be detected in either the spleens or cervical lymph nodes of mice 4, 5, 6, or 7 days after intracranial infection, suggesting that the CTL response to JHMV infection within the central nervous system may be induced or expanded locally. PMID- 8230430 TI - Characterization of stable Chinese hamster ovary cells expressing wild-type, secreted, and glycosylphosphatidylinositol-anchored human immunodeficiency virus type 1 envelope glycoprotein. AB - We generated Chinese hamster ovary cell lines that stably express wild-type, secreted, and glycosylphosphatidylinositol (GPI)-anchored envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1). The cells expressing wild-type Env (WT cells) express both the precursor gp160 and the mature gp120/gp41 and readily form large syncytia when cocultivated with CD4+ human cells. The cells expressing secreted Env (SEC cells) release 140-kDa precursor and mature 120-kDa envelope glycoproteins into the supernatants. The cells expressing GPI-anchored Env (PI cells) express both 140-kDa precursor and mature gp120/gp41 envelope glycoproteins, which can be released from the cell surface by treatment with phosphatidylinositol-specific phospholipase C (PI-PLC). Both the secreted and PI PLC-released envelope glycoproteins form oligomers that can be detected on nonreducing sodium dodecyl sulfate-polyacrylamide gels. In contrast to the WT cells, the SEC and PI cells do not form syncytia when cocultivated with CD4+ human cells. The availability of cells producing water-soluble oligomers of HIV-1 Env should facilitate studies of envelope glycoprotein structure and function. The WT cells, which readily induce syncytia with CD4+ cells, provide a convenient system for assessing potential fusion inhibitors and for studying the fusion mechanism of the HIV Env glycoprotein. PMID- 8230431 TI - Influence of substrate structure on disintegration activity of Moloney murine leukemia virus integrase. AB - The disintegration activity of Moloney murine leukemia virus (M-MuLV) integrase (IN) was investigated through structural and sequence modifications of a Y substrate that resembles an integration intermediate. The Y substrates, constructed from individual oligonucleotides, contain a single viral long terminal repeat (LTR) joined to a nicked target DNA. Truncation of the double stranded LTR sequences distal to the conserved 5'-CA-3' dinucleotide progressively diminished disintegration activity. M-MuLV IN was also able to catalyze disintegration of a heterologous double-stranded LTR sequence. Significantly, the activity of M-MuLV IN on single-stranded LTR Y substrates was more dependent on the sequence and length of the LTR strand than that reported for human immunodeficiency virus type 1 (HIV-1) IN. Modifications introduced at the Y-substrate junction demonstrated that the 3'-hydroxyl group at the terminus of the target strand was necessary for efficient joining of the target DNA strands. The presence of a 2'-hydroxyl group at the 3' end of the target strand, as well as a single-nucleotide gap at the LTR-target junction, reduced disintegration activity. The absence of hydroxyl groups on the terminal nucleotide abolished joining of the target strands. The results presented here suggest that M-MuLV IN disintegration activity is dependent on substantially different LTR sequence requirements than those reported for HIV-1 IN and may be mediated primarily through a structural recognition event. PMID- 8230432 TI - Processing of pestivirus polyprotein: cleavage site between autoprotease and nucleocapsid protein of classical swine fever virus. AB - The polyprotein of classical swine fever virus starts with the nonstructural protein p23, which is followed by the nucleocapsid protein p14. Proteolytic cleavage between p23 and p14 was demonstrated in a cell-free transcription translation system. Successive truncation of the cDNA used for the transcription indicated that the proteolytic activity responsible for the cleavage between p23 and p14 resides within p23. In order to determine the cleavage site between these two proteins, the respective genomic regions were expressed in two different expression systems. N-terminal sequencing of the resulting p14-related proteins revealed that cleavage occurs between Cys-168 and Ser-169. Comparison of the sequence around the cleavage site with sequences of other pestiviruses suggests a conserved processing site between similar proteins. PMID- 8230433 TI - Pathogenicity of molecularly cloned bovine leukemia virus. AB - To delineate the mechanisms of bovine leukemia virus (BLV) pathogenesis, four full-length BLV clones, 1, 8, 9, and 13, derived from the transformed cell line FLK-BLV and a clone construct, pBLV913, were introduced into bovine spleen cells by microinjection. Microinjected cells exhibited cytopathic effects and produced BLV p24 and gp51 antigens and infectious virus. The construct, pBLV913, was selected for infection of two sheep by inoculation of microinjected cells. After 15 months, peripheral blood mononuclear cells from these sheep served as inocula for the transfer of infection to four additional sheep. All six infected sheep seroconverted to BLV and had detectable BLV DNA in peripheral blood mononuclear cells after amplification by polymerase chain reaction. Four of the six sheep developed altered B/T-lymphocyte ratios between 33 and 53 months postinfection. One sheep died of unrelated causes, and one remained hematologically normal. Two of the affected sheep developed B lymphocytosis comparable to that observed in animals inoculated with peripheral blood mononuclear cells from BLV-infected cattle. This expanded B-lymphocyte population was characterized by elevated expression of B-cell surface markers, spontaneous blastogenesis, virus expression in vitro, and increased, polyclonally integrated provirus. One of these two sheep developed lymphocytic leukemia-lymphoma at 57 months postinfection. Leukemic cells had the same phenotype and harbored a single, monoclonally integrated provirus but produced no virus after in vitro cultivation. The range in clinical response to in vivo infection with cloned BLV suggests an important role for host immune response in the progression of virus replication and pathogenesis. PMID- 8230434 TI - Amplification and sequence analysis of DNA flanking integrated proviruses by a simple two-step polymerase chain reaction method. AB - We describe a two-step polymerase chain reaction method that can be used for the amplification of cellular DNA sequences adjacent to an integrated retroviral provirus. The technique involves a partly degenerate, arbitrary primer that will hybridize in the provirus-flanking cellular DNA. By using this primer in combination with a biotinylated provirus-specific primer, a provirus-cellular DNA junction fragment can be isolated from the nonspecific amplification products by using streptavidin-coated magnetic beads. A second amplification employing a nested provirus-specific primer and a biotinylated nondegenerate primer derived from the partly degenerate primer followed by purification with streptavidin coated beads enhances the specificity and the efficiency of recovery of a fragment(s) containing the unknown flanking sequences. In addition to being relevant in studies of viral integration sites, the method should be generally useful to analyze DNA sequences either upstream or downstream from a known sequence. PMID- 8230435 TI - Two E2 binding sites (E2BS) alone or one E2BS plus an A/T-rich region are minimal requirements for the replication of the human papillomavirus type 11 origin. AB - Human papillomaviruses (HPVs) cannot be propagated in vitro, but the DNA can be replicated transiently in an assay in the presence of two trans-acting viral proteins, E1 and E2. Using this assay, we have defined the minimal cis-acting elements of the origin of replication of HPV type 11. Most HPV genomes are conserved at the origin of replication, and the core contains three E2 binding sites (E2BS) surrounding an A/T-rich spacer region. The present results show that the minimal requirement for replication is either two E2BS alone or the A/T-rich region plus one E2BS; in the latter case the relative position of the E2BS is important. In all the studies, the presence of both E1 and E2 proteins was essential for replication, yet only the E2BS was required at the origin. We have shown that E1, E2, and the origin of replication containing an E2BS from a complex in vitro, and our data are consistent with a model in which E2 acts to target E1 to the HPV type 11 replication origin. PMID- 8230436 TI - Escape from in vivo restriction of Moloney mink cell focus-inducing viruses driven by the Mo+PyF101 long terminal repeat (LTR) by LTR alterations. AB - Mo+PyF101 M-MuLV is a variant Moloney murine leukemia virus containing polyomavirus F101 enhancers inserted just downstream from the M-MuLV enhancers in the long terminal repeat (LTR). The protein coding sequences for this virus are identical to those of M-MuLV. Mo+PyF101 M-MuLV induces T-cell disease with a much lower incidence and longer latency than wild-type M-MuLV. We have previously shown that Mo+PyF101 M-MuLV is defective in preleukemic events induced by wild type M-MuLV, including splenic hematopoietic hyperplasia, bone marrow depletion, and generation of recombinant mink cell focus-inducing viruses (MCFs). We also showed that an M-MCF virus driven by the Mo+PyF101 LTR is infectious in vitro but does not propagate in mice. However, in these experiments, when a pseudotypic mixture of Mo+PyF101 M-MuLV and Mo+PyF101 MCF was inoculated into newborn NIH Swiss mice, they died of T-cell leukemia at times almost equivalent to those induced by wild-type M-MuLV. Tumor DNAs from Mo+PyF101 M-MuLV-Mo+PyF101 MCF inoculated mice were examined by Southern blot analysis. The predominant forms of Mo+PyF101 MCF proviruses in these tumors contained added sequences in the U3 region of the LTR. The U3 regions of representative tumor-derived variant Mo+PyF101 MCFs were cloned by polymerase chain reaction amplification, and sequencing indicated that they had acquired an additional copy of the M-MuLV 75 bp tandem repeat in the enhancer region. NIH 3T3 cell lines infected with altered viruses were obtained from representative Mo+PyF101 M-MuLV-Mo+PyF101 MCF-induced tumors, and mice were inoculated with the recovered viruses. Leukemogenicity was approximately equivalent to that in the original Mo+PyF101 M-MuLV-Mo+PyF101 MCF viral stock. Southern blot analysis on the resulting tumors now predominantly revealed loss of the polyomavirus sequences. These results suggest that the suppressive effects of the PyF101 sequences on M-MuLV-induced disease and potentially on MCF propagation were overcome in two ways: by triplication of the M-MuLV direct repeats and by loss of the polyomavirus sequences. PMID- 8230437 TI - Isolation of a herpes simplex virus type 1 mutant with a deletion in the virion host shutoff gene and identification of multiple forms of the vhs (UL41) polypeptide. AB - The virion host shutoff (vhs) gene (UL41) of herpes simplex virus type 1 (HSV-1) encodes a virion component that induces degradation of host mRNAs and the shutoff of most host protein synthesis. Subsequently, the vhs protein accelerates the turnover of all kinetic classes of viral mRNA. To identify the vhs (UL41) polypeptide within infected cells and virions, antisera raised against a UL41 lacZ fusion protein were used to characterize the polypeptides encoded by wild type HSV-1 and two mutants: vhs1, a previously characterized mutant that lacks detectable virion host shutoff activity, and vhs-delta Sma, a newly constructed mutant containing a deletion of 196 codons from UL41. Two forms of the vhs (UL41) polypeptide were identified in cells infected with the wild-type virus or vhs1. Wild-type HSV-1 produced a major 58-kDa polypeptide, as well as a less abundant 59.5-kDa form of the protein, while vhs1 produced 57- and 59-kDa polypeptides that were approximately equally abundant. Although for either virus, both forms of the protein were phosphorylated, they differed in the extent of phosphorylation. While both vhs polypeptides were found in infected cells, only the faster migrating, less phosphorylated form was incorporated into virions. vhs delta Sma encoded a smaller, 31-kDa polypeptide which, although present in infected cells, was not incorporated into virions. The results identify multiple forms of the vhs (UL41) polypeptide and suggest that posttranslational processing affects its packaging into virions, as well as its ability to induce mRNA degradation. PMID- 8230438 TI - Heterologous C-terminal sequences disrupt transcriptional activation and oncogenesis by p59v-rel. AB - Members of the NF-kappa B/rel family of transcription factors are regulated through a trans association with members of a family of inhibitor proteins, collectively known as I kappa B proteins, that contain five to eight copies of a 33-amino-acid repeat sequence (ankyrin repeat). Certain NF-kappa B/rel proteins are also regulated by cis-acting ankyrin repeat-containing domains. The C terminus of p105NF-kappa B, the precursor of the 50-kDa subunit of NF-kappa B, contains a series of ankyrin repeats; proteolytic removal of this ankyrin domain is necessary for the manifestation of sequence-specific DNA binding and nuclear translocation of the N-terminal product. To investigate the structural requirements important for regulation of different NF-kappa B/rel family members by polypeptides containing ankyrin repeat domains, we have constructed a p59v rel:p105NF-kappa B chimeric protein (p110v-rel-ank). The presence of C-terminal p105NF-kappa B-derived sequences in p110v-rel-ank inhibited nuclear translocation, sequence-specific DNA binding, pp40I kappa B-alpha association, and oncogenic transformation. Sequential truncation of the C-terminal ankyrin domain of p110v-rel-ank resulted in the restoration of nuclear translocation, DNA binding, and pp40I kappa B-alpha association but did not restore the oncogenic properties of p59v-rel. The presence of 67 C-terminal p105NF-kappa B-derived amino acids was sufficient to inhibit both transcriptional activation and oncogenic transformation by p59v-rel. These results support a model in which activation of gene expression by p59v-rel is required for its ability to induce oncogenic transformation. PMID- 8230439 TI - Viral genomes maintained extrachromosomally in hamster polyomavirus-induced lymphomas display a cell-specific replication in vitro. AB - Hamster polyomavirus causes lymphomas when injected into newborn Syrian hamsters. Large amounts of extrachromosomal viral genomes are accumulated in the lymphoma cells. These genomes are characterized by deletions affecting the late coding region as well as a specific part of the noncoding regulatory region. By contrast with wild-type genomes, lymphoma-associated genomes replicate in a lymphoblastoid cell line but not in a fibroblastic cell line. The deletion acts in a cis dominant manner and is the primary determinant of this host-range effect on replication. The boundaries of the regulatory region necessary for viral DNA replication in the two cell contexts have been defined. The regulatory region can be functionally divided in two domains: one domain (distal from the origin of replication) is necessary for viral genome replication in fibroblasts, whereas the other domain (proximal to the origin of replication) is functional only in the lymphoblastoid cell context and contains the sequence specifically conserved in the lymphoma-associated genomes. This sequence harbors a motif recognized by a lymphoblastoid cell-specific trans-acting factor. PMID- 8230440 TI - Bromovirus RNA replication and transcription require compatibility between the polymerase- and helicase-like viral RNA synthesis proteins. AB - The positive-strand RNA bromoviruses encode two nonstructural proteins, 1a and 2a, involved in RNA-dependent RNA replication. These proteins have extensive sequence similarities with methyltransferase, helicase, and polymerase proteins of other plant and animal viruses. 1a and 2a can also form a complex in vitro. To explore whether 1a-2a interaction is required for RNA replication in vivo, we reassorted the 1a and 2a genes from two different bromoviruses, brome mosaic virus (BMV) and cowpea chlorotic mottle virus (CCMV). 1a and 2a were expressed independently of viral replication by using RNA- or DNA-based transient expression, and their in vivo RNA replication activities were tested in protoplasts with BMV and CCMV RNA3 templates. RNA-based transient expression confirmed prior indications that bromovirus RNA replication is more sensitive to reductions in 1a expression than to reductions in 2a expression. DNA-based expression of the homologous combinations of 1a and 2a supported high levels of RNA synthesis, but both 1a-2a heterologous combinations exhibited RNA synthesis defects. The combination of CCMV 1a and BMV 2a did not support detectable synthesis of negative-strand, positive-strand, or subgenomic RNA. The converse combination of BMV 1a and CCMV 2a was preferentially defective in positive-strand and subgenomic RNA accumulation, showing that 1a-2a interaction is involved in these processes in ways distinct from negative-strand RNA synthesis, which was only slightly affected. These results indicate that at least some functions of 1a and 2a operate in a mutually dependent manner in vivo and that the mechanisms of positive- and negative-strand RNA synthesis are differentiated in part by features of such interactions. PMID- 8230441 TI - Specific binding of human immunodeficiency virus type 1 gag polyprotein and nucleocapsid protein to viral RNAs detected by RNA mobility shift assays. AB - Packaging of retroviral genomic RNA during virion assembly is thought to be mediated by specific interactions between the gag polyprotein and RNA sequences (often termed the psi or E region) near the 5' end of the genome. For many retroviruses, including human immunodeficiency virus type 1 (HIV-1), the portions of the gag protein and the RNA that are required for this interaction remain poorly defined. We have used an RNA gel mobility shift assay to measure the in vitro binding of purified glutathione S-transferase-HIV-1 gag fusion proteins to RNA riboprobes. Both the complete gag polyprotein and the nucleocapsid (NC) protein alone were found to bind specifically to an HIV-1 riboprobe. Either Cys His box of NC could be removed without eliminating specific binding to the psi riboprobe, but portions of gag containing only the MA and CA proteins without NC did not bind to RNA. There were at least two binding sites in HIV-1 genomic RNA that bound to the gag polyprotein: one entirely 5' to gag and one entirely within gag. The HIV-1 NC protein bound to riboprobes containing other retroviral psi sequences almost as well as to the HIV-1 psi riboprobe. PMID- 8230442 TI - Mutually exclusive binding of two cellular factors within a critical promoter region of the gene for the IE110k protein of herpes simplex virus. AB - We have examined the cis- and trans-acting factors involved in constitutive transcription of the promoter for the IE110k protein of herpes simplex virus type 1. Our results indicate that while the IE110k gene is activated by Vmw65, it also exhibits very efficient constitutive expression approximating that from the simian virus 40 early enhancer-promoter region. We show that despite the presence of multiple copies of the octamer consensus site which mediate Oct-1 binding and subsequent Vmw65 activation, these upstream sequences have a minor effect on constitutive transcription. By progressive exonuclease digestion and subsequent site-directed mutagenesis of the promoter, we have identified a 15-bp region (termed the EC region), from position -89 to -74, which is required for efficient constitutive expression from the IE110k promoter. We demonstrate that two cellular proteins interact with this region and, by competition and methylation interference analyses, show they have distinct but overlapping sequence requirements for binding. One of these proteins is identified as NF-Y, a CCAAT box-binding factor, which binds an inverted CCAAT box located between positions 71 and -75. The second cellular factor, F2, appears to be novel and binds a region with the sequence CGCGCGGC CAT which overlaps the 3' end of the CCAAT box. The terminal AT of the recognition site for F2 represents, on the opposite strand, the terminal AT of the CCAAT box, and these and adjacent bases are critically required for the binding of both factors. These results together with further competition analysis indicate that these factors bind in a mutually exclusive manner to the EC region. The implications of these results for regulation of expression of the IE110k gene are discussed. PMID- 8230443 TI - Three different cellular proteins bind to complementary sites on the 5'-end positive and 3'-end-negative strands of mouse hepatitis virus RNA. AB - The termini of viral genomic RNA and its complementary strand are important in the initiation of viral RNA replication, which probably involves both viral and cellular proteins. To detect the possible cellular proteins involved in the replication of mouse hepatitis virus RNA, we performed RNA-protein binding studies with RNAs representing both the 5' and 3' ends of the viral genomic RNA and the 3' end of the negative-strand complementary RNA. Gel-retardation assays showed that both the 5'-end-positive- and 3'-end-negative-strand RNA formed an RNA-protein complex with cellular proteins from the uninfected cells. UV cross linking experiments further identified a 55-kDa protein bound to the 5' end of the positive-strand viral genomic RNA and two proteins 35 and 38 kDa in size bound to the 3' end of the negative-strand cRNA. The results of the competition assay confirmed the specificity of this RNA-protein binding. No proteins were found to bind to the 3' end of the viral genomic RNA under the same conditions. The binding site of the 55-kDa protein was mapped within the 56-nucleotide region from nucleotides 56 to 112 from the 5' end of the positive-strand RNA, and the 35 and 38-kDa proteins bound to the complementary region on the negative-strand RNA. The 38-kDa protein was detected only in DBT cells but was not detected in HeLa or COS cells, while the 35-kDa protein was found in all three cell types. The juxtaposition of the different cellular proteins on the complementary sites near the ends of the positive- and negative-strand RNAs suggests that these proteins may interact with each other and play a role in mouse hepatitis virus RNA replication. PMID- 8230444 TI - Rescue of an influenza A virus wild-type PB2 gene and a mutant derivative bearing a site-specific temperature-sensitive and attenuating mutation. AB - Live attenuated influenza A virus vaccines are currently produced by the transfer of attenuating genes from a donor virus to new epidemic variants of influenza A virus, with the selection of reassortant viruses that possess the protective antigens (i.e., the two surface glycoproteins) of the epidemic virus and the attenuating genes from the donor virus. The previously studied attenuated donor viruses were produced by conventional methods such as passage of virus at low temperature or chemical mutagenesis. The present paper describes a new strategy for the generation of a donor virus bearing an attenuating, non-surface glycoprotein gene. This strategy involves the introduction of attenuating mutations into the cDNA copy of the PB2 polymerase gene by site-directed mutagenesis, transfection of in vitro RNA transcripts of PB2 cDNA, and recovery of the transfected PB2 gene into an infectious virus. An avian-human influenza A virus PB2 single-gene reassortant virus (with an avian influenza A virus PB2 gene) that replicates efficiently in avian tissue but poorly in mammalian cells was used as a helper virus to rescue a transfected synthetic RNA derived from a human influenza A virus PB2 gene. The desired human influenza A virus mutant PB2 transfectant was favored in this situation because the avian influenza A virus PB2 gene restricts viral replication in mammalian cells in culture, the system used for rescue, thereby providing strong selection for the virus bearing the human influenza A virus PB2 gene. We validated the feasibility of this approach by rescuing the PB2 gene of the wild-type influenza A/Ann Arbor/6/60 virus and a mutant derivative that had a single amino acid substitution introduced at position 265 by site-directed mutagenesis. Previously, this amino acid substitution had been shown to specify both a temperature-sensitive (ts) and an attenuation (att) phenotype. The rescued mutant 265 PB2 transfectant virus exhibited the ts and att phenotypes, which confirms that these phenotypes were specified by this single amino acid substitution. The transfectant virus was immunogenic and protected hamsters from subsequent challenge with wild-type virus. The cDNA copy of this influenza A/Ann Arbor/6/60 virus mutant 265 PB2 gene will be used as a substrate for the introduction of additional attenuating mutations by site-directed mutagenesis. PMID- 8230445 TI - Incorporation of Vpr into human immunodeficiency virus type 1 virions: requirement for the p6 region of gag and mutational analysis. AB - The product of the vpr open reading frame of human immunodeficiency virus type 1 (HIV-1) is a 15-kDa, arginine-rich protein that is present in virions in molar quantities equivalent to that of Gag. We report here the results of our investigations into the mechanism by which Vpr is incorporated into virions during assembly in infected cells. For these studies we used an expression vector encoding a Vpr molecule fused at its amino terminus to a nine-amino-acid peptide from influenza virus hemagglutinin. The tagged Vpr expression vector and a vpr mutant HIV-1 provirus were used to cotransfect COS cells, and the resulting virions were tested for the presence of the tagged protein on immunoblots probed with monoclonal antibody against the hemagglutinin peptide. The COS-produced virions were found to contain readily detectable amounts of tagged Vpr and smaller amounts of a putative tagged Vpr dimer. Infectivity of the particles was not altered by incorporation of tagged Vpr. Our results using this system in combination with mutant HIV-1 proviruses suggested that incorporation of Vpr into virions requires the carboxy-terminal Gag protein of HIV-1 (p6) but not gp160, Pol, or genomic viral RNA. In addition, analysis of mutated, tagged Vpr molecules suggested that amino acids near the carboxy terminus (amino acids 84 to 94) are required for incorporation of Vpr into HIV-1 virions. The single cysteine residue near the carboxy terminus was required for production of a stable protein. Arginine residues tested were not important for incorporation or stability of tagged Vpr. These results suggested a novel strategy for blocking HIV-1 replication. PMID- 8230446 TI - Vpu-induced degradation of CD4: requirement for specific amino acid residues in the cytoplasmic domain of CD4. AB - Two functions have been attributed to the product of the human immunodeficiency virus type 1 vpu open reading frame: it increases virion release from infected cells and induces rapid degradation of CD4 shortly after its synthesis. In the absence of Vpu, newly synthesized gp160 and CD4 associate in the endoplasmic reticulum (ER), forming a complex whose further maturation is blocked and which is eventually degraded. In studies using NL4-3-based expression vectors, it has been previously shown that Vpu induces the release of gp160 from the complex that it forms with CD4 in the ER. This release, which appears to be due to the rapid degradation of CD4 induced by Vpu, allows gp160 to transit to the Golgi, where it matures further. We investigated which regions of CD4 are important for its susceptibility to Vpu-induced degradation by transfecting HeLa cells with isogenic vpu-positive and vpu-negative proviruses and vectors expressing various truncated or mutated CD4 molecules. The results suggested that the cytoplasmic domain of CD4 contains a determinant lying within amino acids 418 to 425 that is critical for susceptibility to Vpu-induced degradation. Neither the phosphorylation sites in the cytoplasmic domain nor the Lck interaction region was required for the effect. Vpu-induced degradation was specific for CD4, since CD8, even when retained in the ER, was not degraded. In addition, under conditions of high-level Vpu expression, CD4 degradation could be observed in the absence of gp160 or other means of retaining CD4 in the ER. PMID- 8230447 TI - Vesicular stomatitis virus M protein may be inside the ribonucleocapsid coil. AB - Vesicular stomatitis virus is an enveloped virus with an external glycoprotein G and a nucleocapsid that form, together with the M protein, a tight helically coiled structure: the skeleton. Negative staining and immunoelectron microscopy studies on skeleton preparations were performed to determine the localization of the M protein. These studies have resulted in a new model for the structure of rhabdoviruses in which the nucleocapsid is wound around a core containing the M protein. This model predicts contact between M and lipid only at the extreme ends of the skeleton, which is confirmed by skeleton-liposome binding studies. PMID- 8230448 TI - Binding sites for the herpes simplex virus immediate-early protein ICP4 impose an increased dependence on viral DNA replication on simple model promoters located in the viral genome. AB - We examined the ability of binding sites for the herpes simplex virus immediate early protein ICP4 to alter the regulation of closely linked promoters by placing strong ICP4 binding sites upstream or downstream of simple TATA promoters in the intact viral genome. We found that binding sites strongly reduced the levels of expression at early times postinfection and that this effect was partially overcome after the onset of viral DNA replication. These data confirm that DNA bound ICP4 can inhibit the activity of a closely linked promoter and raise the possibility that ICP4 binding sites contribute to temporal regulation during infection. PMID- 8230449 TI - Purification of the late transcription system of vaccinia virus: identification of a novel transcription factor. AB - We have resolved the in vitro late transcription system of vaccinia virus into four components consisting of RNA polymerase and three accessory factors. One of these additional factors is a 30-kDa protein which was previously shown to be required for late transcription in vitro and was indirectly shown to be the product of the G8R open reading frame. Another factor, of 17 kDa, was previously identified as a possible late transcription factor by an assay which demonstrated that the gene encoding it, A1L, was required for late gene expression in vivo. The G8R and A1L open reading frames have now been cloned into a baculovirus expression system, and the corresponding proteins have been purified. Both are necessary for late transcription in vitro, confirming that these intermediate genes encode late transcription factors. The third factor has a sedimentation coefficient consistent with a protein of 32 to 38 kDa. Experimental results suggest that this is a previously unidentified factor encoded by a vaccinia virus early gene. The RNA polymerase functioning in this system was purified from vaccinia virus-infected cells; however, it can be complemented by the RNA polymerase which is packaged in virions. The three smaller proteins and RNA polymerase are all necessary, and together are sufficient, for the synthesis of late viral mRNA in vitro. PMID- 8230450 TI - Human immunodeficiency virus type 1 pol gene mutations in an AIDS patient treated with multiple antiretroviral drugs. AB - Multiple mutations were found in the human immunodeficiency virus pol gene following treatment of an AIDS patient with antiretroviral drugs. After approximately 2.5 years of monthly alternating therapy with 3'-azido-3' deoxythymidine (AZT) and 2',3'-dideoxycytidine (ddC), most of the pol sequences amplified from the patient's peripheral blood mononuclear cell DNA contained known AZT resistance mutations at codons 41, 67, and 215 and a putative ddC resistance mutation at codon 69 as well as other novel mutations. These mutations persisted for 6 months after the patient was switched to 2',3'-dideoxyinosine monotherapy. Mutations known to be associated with 2',3'-dideoxyinosine resistance did not occur during this time. Antiviral susceptibility testing of point mutants, introduced into the genetic background of laboratory strain NL4-3, showed that the codon 41 mutation antagonized ddC resistance when present with the codon 69 mutation. However, this antagonism was not found with a chimeric mutant containing the patient's pol gene sequence from codons 25 to 218, implying that other mutations compensated for the antagonism. Thus, alternating therapy with AZT and ddC resulted in the selection of viruses resistant to both drugs. PMID- 8230451 TI - Herpes simplex virus type 1 latency-associated transcript (LAT) promoter deletion mutants can express a 2-kilobase transcript mapping to the LAT region. AB - The results of studies in several laboratories suggest that a TATA box-containing promoter located in the herpes simplex virus type 1 internal long repeat and long terminal repeat elements drives expression of the latency-associated transcripts (LATs). In the present study, we show that expression of a 2-kb LAT-related transcript can occur in the absence of this LAT TATA promoter, indicating the existence of a cryptic promoter. By Northern (RNA) blot analysis, we have examined LAT expression by herpes simplex virus type 1 variant strains KOS/29 and 1704, which contain deletions of the LAT promoter region. Our data indicate that KOS/29, despite lacking the 203-bp fragment which contains the LAT TATA box, can express a 2-kb LAT-related transcript during productive infection in tissue culture and in mouse trigeminal ganglia during acute infection and reactivation. Similarly, strain 1704, which contains a larger deletion in this promoter region, also expresses a 2-kb LAT-related transcript during tissue culture infection and reactivation of latently infected trigeminal ganglia. However, LATs are not expressed with either virus during latency. Northern blot analysis with a single stranded, oligonucleotide probe demonstrates that the 2-kb LAT and LAT-related transcript are colinear and share a large area of sequence similarity. These findings suggest the existence of a second promoter in the LAT gene which can function during lytic infection and reactivation, at least in the absence of the LAT TATA promoter. We propose that this cryptic promoter is located either in a proximal region approximately 300 bp upstream of the start site of the 2-kb LAT or in a distal region starting over 1,226 bp upstream of this site. PMID- 8230452 TI - Poliovirus capsid proteins derived from P1 precursors with glutamine-valine cleavage sites have defects in assembly and RNA encapsidation. AB - Assembly of poliovirus virions requires proteolytic cleavage of the P1 capsid precursor polyprotein between two separate glutamine-glycine (QG) amino acid pairs by the viral protease 3CD. In this study, we have investigated the effects on P1 polyprotein processing and subsequent assembly of processed capsid proteins caused by substitution of the glycine residue at the individual QG cleavage sites with valine (QG-->QV). P1 cDNAs encoding the valine substitutions were created by site-directed mutagenesis and were recombined into wild-type vaccinia virus to generate recombinant vaccinia viruses which expressed the mutant P1 precursors. The recombinant vaccinia virus-expressed mutant P1 polyproteins were analyzed for proteolytic processing defects in cells coinfected with a recombinant vaccinia virus (VVP3) that expresses the poliovirus 3CD protease and for processing and assembly defects by using a trans complementation system in which P1-expressing recombinant vaccinia viruses provide capsid precursor to a defective poliovirus genome that does not express functional capsid proteins (D. C. Ansardi, D. C. Porter, and C. D. Morrow, J. Virol. 67:3684-3690, 1993). The QV-substituted precursors were proteolytically processed at the altered sites both in cells coinfected with VVP3 and in cells coinfected with defective poliovirus, although the kinetics of cleavage at the altered sites were slower than those of cleavage at the wild-type QG site in the precursor. Completely processed capsid proteins VP0, VP3, and VP1 derived from the mutant precursor containing a valine at the amino terminus of VP3 (VP3-G001V) were unstable and failed to assemble stable subviral structures in cells coinfected with defective poliovirus. In contrast, capsid proteins derived from the P1 precursor with a valine substitution at the amino terminus of VP1 (VP1-G001V) assembled empty capsid particles but were deficient in assembling RNA-containing virions. The assembly characteristics of the VP1-G001V mutant were compared with those of a previously described VP3-VP1 cleavage site mutant (K. Kirkegaard and B. Nelsen, J. Virol. 64:185-194, 1990) which contained a deletion of the first four amino-terminal residues of VP1 (VP1 delta 1-4) and which was reconstructed for our studies into the recombinant vaccinia virus system. Complete proteolytic processing of the VP1-delta 1-4 precursor also occurred more slowly than complete cleavage of the wild-type precursor, and formation of virions was delayed; however, capsid proteins derived from the VP1-G001V mutant assembled RNA-containing virions less efficiently than those derived from the VP1-delta 1-4 precursor.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8230453 TI - Epstein-Barr virus recombinants from overlapping cosmid fragments. AB - Five overlapping type 1 Epstein-Barr virus (EBV) DNA fragments constituting a complete replication- and transformation-competent genome were cloned into cosmids and transfected together into P3HR-1 cells, along with a plasmid encoding the Z immediate-early activator of EBV replication. P3HR-1 cells harbor a type 2 EBV which is unable to transform primary B lymphocytes because of a deletion of DNA encoding EBNA LP and EBNA 2, but the P3HR-1 EBV can provide replication functions in trans and can recombine with the transfected cosmids. EBV recombinants which have the type 1 EBNA LP and 2 genes from the transfected EcoRI A cosmid DNA were selectively and clonally recovered by exploiting the unique ability of the recombinants to transform primary B lymphocytes into lymphoblastoid cell lines. PCR and immunoblot analyses for seven distinguishing markers of the type 1 transfected DNAs identified cell lines infected with EBV recombinants which had incorporated EBV DNA fragments beyond the transformation marker-rescuing EcoRI-A fragment. Approximately 10% of the transforming virus recombinants had markers mapping at 7, 46 to 52, 93 to 100, 108 to 110, 122, and 152 kbp from the 172-kbp transfected genome. These recombinants probably result from recombination among the transfected cosmid-cloned EBV DNA fragments. The one recombinant virus examined in detail by Southern blot analysis has all the polymorphisms characteristic of the transfected type 1 cosmid DNA and none characteristic of the type 2 P3HR-1 EBV DNA. This recombinant was wild type in primary B-lymphocyte infection, growth transformation, and lytic replication. Overall, the type 1 EBNA 3A gene was incorporated into 26% of the transformation marker-rescued recombinants, a frequency which was considerably higher than that observed in previous experiments with two-cosmid EBV DNA cotransfections into P3HR-1 cells (B. Tomkinson and E. Kieff, J. Virol. 66:780-789, 1992). Of the recombinants which had incorporated the marker-rescuing cosmid DNA fragment and the fragment encoding the type 1 EBNA 3A gene, most had incorporated markers from at least two other transfected cosmid DNA fragments, indicating a propensity for multiple homologous recombinations. The frequency of incorporation of the nonselected transfected type 1 EBNA 3C gene, which is near the end of two of the transfected cosmids, was 26% overall, versus 3% in previous experiments using transfections with two EBV DNA cosmids. In contrast, the frequency of incorporation of a 12-kb EBV DNA deletion which was near the end of two of the transfected cosmids was only 13%.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8230454 TI - Transcriptional activation of the human T-lymphotropic virus type I long terminal repeat by functional interaction of Tax1 and Ets1. AB - Transcription regulation of the oncogenic retrovirus human T-lymphotropic virus type I (HTLV-I) involves the composite activity of both viral and cellular transcription factors. The HTLV-I transforming protein, Tax1, modulates the activity of several cellular transcription factors, upregulating the level of viral gene expression. In addition, cellular transcription factors, such as Ets1, independently bind to the viral long terminal repeat in a sequence-specific manner and activate transcription. It was of interest to analyze the possible interaction of Tax1 and Ets1 in viral gene regulation. We now report that Tax1 and Ets1 increase expression from the HTLV-I promoter in a cooperative manner. The level of expression was increased 5- to 10-fold above the combined individual effect of Tax1 and Ets1. S1 nuclease analysis demonstrated that the cooperative effect was due to an increase in the levels of steady-state RNA. The functional interaction between Tax1 and Ets1 required the presence of the Tax1-responsive 21 bp repeat element TRE-1 and the Ets1-responsive element ERR-1. These results suggested the possible interaction of Ets1 with transcriptional regulatory proteins that bind to the 21-bp repeats. This interaction is demonstrated by decreased electrophoretic mobility of specific 21-bp repeat gel shift complexes in the presence of Ets1. Furthermore, interaction of Ets1 with the 21-bp repeat binding proteins enhances the relative efficiency of binding to the DNA. This cooperative interaction between Ets1 and proteins which bind to the Tax1 responsive 21-bp repeats suggests a possible role for Ets1 in the regulation of viral gene expression. PMID- 8230455 TI - Identification of the activation domain of equine infectious anemia virus rev. AB - Several members of the lentivirus family of complex retroviruses have been shown to encode proteins that are functionally equivalent to the Rev posttranscriptional regulatory protein of human immunodeficiency virus type 1 (HIV-1). Furthermore, the domain organization of HIV-1 Rev, featuring a highly basic N-terminal RNA binding domain and a leucin-rich C-terminal effector domain, has also been shown to be highly conserved among Rev proteins derived from not only the primate but also the ovine and caprine lentiviruses. Although it has therefore appeared highly probable that the lentivirus equine infectious anemia virus (EIAV) also encodes a Rev, the predicted amino acid sequence of this putative EIAV regulatory protein does not display any evident homology to the basic and leucine-rich motifs characteristic of other known Rev proteins. By fusion of different segments of the proposed EIAV Rev protein to the well-defined RNA binding domain of either HIV-1 or visna virus Rev, we have identified a segment of this EIAV protein that can efficiently substitute in cis for the otherwise essential activation motif. Interestingly, the minimal EIAV Rev activation motif identified in this study comprises approximately 18 amino acids located toward the protein N terminus that lack any evident similarity to the leucine-rich activation domains found in these other lentivirus Rev proteins. It therefore appears that the Rev protein of EIAV, while analogous in function to Rev proteins defined in lentiviruses of primate, ovine, and caprine origin, is nevertheless distinguished by an entirely novel domain organization. PMID- 8230456 TI - Targeted mutagenesis of simian virus 40 DNA mediated by a triple helix-forming oligonucleotide. AB - Triple-helical DNA can be formed by oligonucleotides that bind as third strands of DNA in a sequence-specific manner in the major groove in homopurine/homopyrimidine stretches in duplex DNA. Such triple helix-forming oligonucleotides have been used to inhibit gene expression by blocking transcription factor access to promoter sites in transient expression assays. In an alternative approach to genetic manipulation using triplex DNA, we show that triplex-forming oligonucleotides can be used to produce site-specific, targeted mutations in a viral genome in order to achieve a permanent, heritable effect on gene function and expression. We use a triplex-forming oligonucleotide linked to a psoralen derivative at its 5' end to achieve targeted mutagenesis in a simian virus 40 (SV40) vector genome. Site-specific triplex formation delivers the psoralen to the targeted site in the SV40 DNA. Photoactivation of the psoralen yields adducts and thereby mutations at that site. Mutations were produced in the target gene in over 6% of the viral genomes. DNA sequence analysis of the mutations in the target gene showed that all were in the targeted region, and 55% were found to be the same T:A-to-A:T transversion precisely at the targeted base pair. In control experiments, no mutagenesis above the background frequency in the assay was produced by a non-triplex-forming, psoralen-linked oligonucleotide unless a vast excess of this oligonucleotide was used, demonstrating the specificity of the targeted mutagenesis. This frequency of targeted mutagenesis of SV40 in monkey cells represents a 30-fold increase relative to similar experiments using lambda phage in bacteria, suggesting that fixation of the triplex-directed lesion into a mutation occurs more efficiently in mammalian cells. If the ability to reproducibly and predictably target mutations to sites in viral DNA in vitro by using modified oligonucleotides can be extended to DNA in vivo, this approach may prove useful as a technique for gene therapy, as a strategy for antiviral therapeutics, and as a tool for genetic engineering. PMID- 8230457 TI - Individual repeats of Drosophila Myb can function in transformation by v-Myb. AB - The v-Myb protein binds to specific DNA sequences and can regulate gene expression. The DNA-binding domain of v-Myb contains the second and third of the three highly conserved tandem repeats found in c-Myb. In general, the ability of mutant forms of v-Myb to transform correlates with their ability to trans activate transcription. Two mutations within the DNA-binding domain of v-Myb which preserve DNA binding in vitro but fail to trans activate or transform have been described. These results suggested that this highly conserved domain might function in specific protein-protein interactions, as well as in DNA binding. We therefore tested the ability of a related protein domain from Drosophila melanogaster to substitute functionally for the homologous region of v-Myb. We found that either the second or third repeat of Drosophila Myb, but not both, could function in trans-activation and transformation by v-Myb. The hybrid containing both the second and third repeats of Drosophila Myb bound to DNA but failed to trans activate transcription either in the context of v-Myb or as a v Myb-VP16 fusion protein. These results demonstrate that although the protein-DNA contacts made by the Myb repeats have been conserved during the evolution of animals, the protein-protein interactions have diverged. PMID- 8230458 TI - Replication of lymphocytic choriomeningitis virus is restricted in terminally differentiated neurons. AB - We have investigated the replication of lymphocytic choriomeningitis virus (LCMV) before and after the nerve growth factor (NGF)-induced transdifferentiation of PC12 cells from the chromaffin to the neuron-like phenotype. Untreated and NGF treated cells were equally susceptible to LCMV infection; however, the viral yield was found to be 1,000-fold lower in NGF-differentiated PC12 cells. The reduced viral yield correlated with restricted LCMV replication and transcription within the infected cell, which was not caused by the lack of cell proliferation in the NGF-treated cells but rather was related to the induction or changes in expression levels of specific gene product(s) associated with the cell commitment to a neuronal phenotype. The return to the chromaffin phenotype after withdrawal of NGF restored normal LCMV yields as well as levels of viral replication and transcription. The finding of reduced viral replication in terminally differentiated neuronal cells has important implications for understanding the mechanism by which neurotropic viruses, such as LCMV, are able to establish a long-term persistent infection in the central nervous system in the absence of severe pathological changes. PMID- 8230459 TI - Herpesvirus proteinase: site-directed mutagenesis used to study maturational, release, and inactivation cleavage sites of precursor and to identify a possible catalytic site serine and histidine. AB - The cytomegalovirus maturational proteinase is synthesized as a precursor that undergoes at least three processing cleavages. Two of these were predicted to be at highly conserved consensus sequences--one near the carboxyl end of the precursor, called the maturational (M) site, and the other near the middle of the precursor, called the release (R) site. A third less-well-conserved cleavage site, called the inactivation (I) site, was also identified near the middle of the human cytomegalovirus 28-kDa assemblin homolog. We have used site-directed mutagenesis to verify all three predicted sequences in the simian cytomegalovirus proteinase, and have shown that the proteinase precursor is active without cleavage at these sites. We have also shown that the P4 tyrosine and the P2 lysine of the R site were more sensitive to substitution than the other R- and M site residues tested: substitution of alanine for P4 tyrosine at the R site severely reduced cleavage at that site but not at the M site, and substitution of asparagine for lysine at P2 of the R site reduced M-site cleavage and nearly eliminated I-site cleavage but had little effect on R-site cleavage. With the exception of P1' serine, all R-site mutations hindered I-site cleavage, suggesting a role for the carboxyl end of assemblin in I-site cleavage. Pulse chase radiolabeling and site-directed mutagenesis indicated that assemblin is metabolically unstable and is degraded by cleavage at its I site. Fourteen amino acid substitutions were also made in assemblin, the enzymatic amino half of the proteinase precursor. Among those tested, only 2 amino acids were identified as essential for activity: the single absolutely conserved serine and one of the two absolutely conserved histidines. When the highly conserved glutamic acid (Glu22) was substituted, the proteinase was able to cleave at the M and I sites but not at the R site, suggesting either a direct (e.g., substrate recognition) or indirect (e.g., protein conformation) role for this residue in determining substrate specificity. PMID- 8230460 TI - Disulfide bonds in folding and transport of mouse hepatitis coronavirus glycoproteins. AB - We have analyzed the effects of reducing conditions on the folding of the spike (S) protein and on the intracellular transport of the membrane (M) protein of the mouse hepatitis coronavirus. These proteins differ in their potential to form disulfide bonds in the lumen of the endoplasmic reticulum (ER). Intrachain disulfide bonds are formed in the S protein but not in M, which was demonstrated in a pulse-chase experiment by analyzing the viral proteins under nonreducing conditions. To reduce disulfide bonds in vivo, we added dithiothreitol (DTT) to the culture medium of mouse hepatitis coronavirus-infected cells following a procedure recently described by Braakman et al. (I. Braakman, J. Helenius, and A. Helenius, EMBO J. 11:1717-1722, 1992). Short exposure to DTT resulted in the complete reduction of newly synthesized S protein and affected its conformation as judged by the change in mobility in nonreducing gels and by the loss of recognition by a conformation-specific monoclonal antibody. Using this antibody in an immunofluorescence assay, we monitored the reducing effect of DTT in situ. DTT was found to initially affect only the S protein present in the ER; also, after longer treatment, the remaining signal also gradually disappeared. In contrast, folding and transport of the M protein were not inhibited by DTT. Under reducing conditions, M was transported efficiently to the trans side of the Golgi complex, indicating that cellular processes such as ER-to-Golgi transport, O glycosylation, and Golgi retention were unaffected. In the presence of DTT, the M protein even moved at an increased rate to the Golgi complex, which is probably because of its failure to interact with unfolded S protein. The effects of in vivo reduction were reversible. When DTT was removed from pulse-labeled cells, the S protein folded posttranslationally and aberrantly; during its oxidation, most of S now transiently aggregated into large disulfide-linked complexes from which subsequently folded S molecules dissociated. PMID- 8230461 TI - Sequences determining the pH dependence of viral entry are distinct from the host range-determining region of the murine ecotropic and amphotropic retrovirus envelope proteins. AB - The entry of ecotropic and amphotropic murine leukemia retroviruses (MuLV) into cells was investigated by using viral vector particles carrying chimeric amphotropic-ecotropic envelope glycoproteins on their surface. Chimeras were made by joining, at or near the polyproline hinge, the N-terminal portion of the amphotropic (4070A) gp70 onto the C-terminal portion of the ecotropic (Moloney) gp70 and p15E (constructs AE2 and AE4) or vice versa (AE12). Transduction efficiency of the constructs was tested on target cells that either have only ecotropic receptors (CHO-2 and CHO-11 cells), only amphotropic receptors (mink lung fibroblasts and Cos 1 cells), or both types of receptors (NIH 3T3 cells). The assay made use of the fact that the mechanism for viral entry of ecotropic viruses is pH dependent while that of amphotropic viruses is pH independent. Treatment of target cells with NH4Cl, which prevents the reduction of pH within endosomes, reduced the titers of viral particles bearing the C-terminal moiety from the ecotropic envelope but did not reduce the titers of particles which had a C-terminal moiety from the amphotropic envelope. In addition, in contrast to other low-pH-dependent enveloped viruses, brief acid treatment did not allow surface-bound viruses to bypass the NH4Cl block. The results indicate that the pH dependence of viral entry is a property of the sequences C terminal to the polyproline hinge. PMID- 8230462 TI - Hepadnavirus infection requires interaction between the viral pre-S domain and a specific hepatocellular receptor. AB - To better define the molecules involved in the initial interaction between hepadnaviruses and hepatocytes, we performed binding and infectivity studies with the duck hepatitis B virus (DHBV) and cultured primary duck hepatocytes. In competition experiments with naturally occurring subviral particles containing DHBV surface proteins, these DNA-free particles were found to interfere with viral infectivity if used at sufficiently high concentrations. In direct binding saturation experiments with radiolabelled subviral particles, a biphasic titration curve containing a saturable component was obtained. Quantitative evaluation of both the binding and the infectivity data indicates that the duck hepatocyte presents about 10(4) high-affinity binding sites for viral and subviral particles. Binding to these productive sites may be preceded by reversible virus attachment to a large number of less specific, nonsaturable primary binding sites. To identify which of the viral envelope proteins is responsible for hepatocyte-specific attachment, subviral particles containing only one of the two DHBV surface proteins were produced in Saccharomyces cerevisiae. In infectivity competition experiments, only particles containing the large pre-S/S protein were found to markedly reduce the efficiency of DHBV infection, while particles containing the small S protein had only a minor effect. Similarly, physical binding of radiolabelled serum-derived subviral particles to primary duck hepatocytes was inhibited well only by the yeast derived pre-S/S particles. Together, these results strongly support the notion that hepadnaviral infection is initiated by specific attachment of the pre-S domain of the large DHBV envelope protein to a limited number of hepatocellular binding sites. PMID- 8230463 TI - High viral load in lymph nodes and latent human immunodeficiency virus (HIV) in peripheral blood cells of HIV-1-infected chimpanzees. AB - We have examined human immunodeficiency virus type 1 (HIV-1) infection in chimpanzees by analyzing HIV-1 DNA and RNA in lymph nodes and peripheral mononuclear cells (PBMCs). Like certain asymptomatic HIV-infected persons, these chimpanzees had no detectable viral replication in their PBMCs. However, viral replication and a high viral load were observed in the lymphatic tissue. Despite the absence of viral replication in PBMCs, 1/1,000 to 1/10,000 of the PBMCs contained HIV-1 proviral DNA, and HIV transcription could be rapidly induced in these cells in vitro. These results provide direct evidence of cellular latency of HIV in vivo and suggest that HIV infection in chimpanzees may be a useful model for clinical latency of HIV infection in humans. PMID- 8230464 TI - The hemagglutinins of duck and human H1 influenza viruses differ in sequence conservation and in glycosylation. AB - We determined the deduced amino acid sequences of two H1 duck influenza A virus hemagglutinins (HAs) and found that the consensus sequence of the HA, determined directly from virus recovered from the intestinal tract, remains unchanged through many generations of growth in MDCK cells and chicken embryos. These two duck viruses differ from each other by 5 amino acids and from A/Dk/Alberta/35/1976 (F. J. Austin, Y. Kawaoka, and R. G. Webster, J. Gen. Virol. 71:2471-2474, 1990) by 9 and 12 amino acids, most of which are in the HA1 subunit. They are antigenically similar to each other but different from the Alberta virus. We compared these H1 duck HAs with the HAs of human isolates to identify structural properties of this viral glycoprotein that are associated with host range. By comparison to the human H1 HAs, the duck virus HA sequences are highly conserved as judged by the small fraction of nucleotide differences between strains which result in amino acid substitutions. However, the most striking difference between these duck and human HAs is in the number and distribution of glycosylation sites. Whereas duck and swine viruses have four and five conserved glycosylation sites per HA1 subunit, none of which are on the tip of the HA, all human viruses have at least four additional sites, two or more of which are on the tip of the HA. These findings stress the role of glycosylation in the control of host range and suggest that oligosaccharides on the tip of the HA are important to the survival of H1 viruses in humans but not in ducks or swine. PMID- 8230465 TI - Interleukin-2 downregulates hepatitis B virus gene expression in transgenic mice by a posttranscriptional mechanism. AB - We have recently demonstrated that tumor necrosis factor alpha (TNF-alpha) and interleukin-2 (IL-2) downregulate the hepatic steady-state content of hepatitis B virus (HBV) mRNA in vivo in HBV-transgenic mice and that the IL-2 effect is mediated by TNF-alpha. In the current study, we demonstrate that IL-2-induced downregulation of hepatic HBV 2.1-kb mRNA is not due to changes in the transcription rate or the intranuclear maturation or export of this transcript but that it is selectively and profoundly depleted from the cytoplasm of the liver cells in vivo following IL-2 administration. Collectively, these results suggest that IL-2 alters the steady-state content of hepatic HBV mRNA by a posttranscriptional mechanism in vivo, that this effect is mediated by TNF-alpha, and that it probably reflects increased cytoplasmic degradation of the viral transcript. PMID- 8230466 TI - In vivo footprinting analysis of constitutive and inducible protein-DNA interactions at the long terminal repeat of human immunodeficiency virus type 1. AB - The regulation of the rate of transcription of human immunodeficiency virus type 1 is mainly exerted through the long terminal repeat (LTR) at the 5' end of the provirus. A large number of cis-acting regulatory elements have been identified in the LTR by in vitro binding studies; the biological role of these sites within living infected cells, however, is still not clear. We have studied the interactions of nuclear proteins with the LTR in the U1 monocytic cell line by in vivo dimethylsulfate footprinting, using the ligation-mediated polymerase chain reaction technique. In this cell line, transcription of the virus, which is very low under basal conditions, is highly inducible by treatment with phorbol esters; therefore, this system is likely to represent a suitable cellular model to study viral latency. Independently of the level of viral transcription, major in vivo footprints appear at the two Sp1 sites adjacent to the enhancer, the downstream positioned enhancer repeat, the NFAT binding site, and one of the purine-rich sites of the negative regulatory element. Upon transcriptional activation by phorbol myristate acetate, the only perturbation in the footprinting pattern is a dramatic increase in dimethylsulfate sensitivity of guanine at position -92 in the downstream enhancer repeat. This modification is correlated with the transient induction of two enhancer-binding activities, as determined by gel retardation assays. While the transcriptional rate is still increasing and the in vivo footprinting pattern is unchanged at up to 24 h postactivation, these enhancer-binding factors are considerably reduced at this time. Therefore, further levels of regulation have to be considered to explain the maintenance of the induced state. PMID- 8230467 TI - Minimum internal ribosome entry site required for poliovirus infectivity. AB - Translation initiation by internal ribosome binding is a recently discovered mechanism of eukaryotic viral and cellular protein synthesis in which ribosome subunits interact with the mRNAs at internal sites in the 5' untranslated RNA sequences and not with the 5' methylguanosine cap structure present at the extreme 5' ends of mRNA molecules. Uncapped poliovirus mRNAs harbor internal ribosome entry sites (IRES) in their long and highly structured 5' noncoding regions. Such IRES sequences are required for viral protein synthesis. In this study, a novel poliovirus was isolated whose genomic RNA contains two gross deletions removing approximately 100 nucleotides from the predicted IRES sequences within the 5' noncoding region. The deletions originated from previously in vivo-selected viral revertants displaying non-temperature-sensitive phenotypes. Each revertant had a different predicted stem-loop structure within the 5' noncoding region of their genomic RNAs deleted. The mutant poliovirus (Se1 5NC-delta DG) described in this study contains both stem-loop deletions in a single RNA genome, thereby creating a minimum IRES. Se1-5NC-delta DG exhibited slow growth and a pinpoint plaque phenotype following infection of HeLa cells, delayed onset of protein synthesis in vivo, and defective initiation during in vitro translation of the mutated poliovirus mRNAs. Interestingly, the peak levels of viral RNA synthesis in cells infected with Se1-5NC-delta DG occurred at slightly later times in infection than those achieved by wild-type poliovirus, but these mutant virus RNAs accumulated in the host cells during the late phases of virus infection. UV cross-linking assays with the 5' noncoding regions of wild type and mutated RNAs were carried out in cytoplasmic extracts from HeLa cells and neuronal cells and in reticulocyte lysates to identify the cellular factors that interact with the putative IRES elements. The cellular proteins that were cross-linked to the minimum IRES may represent factors playing an essential role in internal translation initiation of poliovirus mRNAs. PMID- 8230468 TI - Comparing regions of the Epstein-Barr virus ZEBRA protein which function as transcriptional activating sequences in Saccharomyces cerevisiae and in B cells. AB - The ZEBRA protein activates expression of Epstein-Barr virus early-lytic-cycle genes in human B lymphocytes. Here it is shown that ZEBRA also behaves as a sequence-specific transcriptional activator in Saccharomyces cerevisiae. Deletional mutagenesis defined three regions of ZEBRA that participate in activation in S. cerevisiae. These regions are designated YI (amino acids [aa] 1 to 25), YII (aa 51 to 102), and YIII (aa 228 to 245). Two of the three regions of the native ZEBRA protein act together to mediate activation when assayed on ZEBRA binding sites. However, when fused to the DNA binding domain of GAL4 and assayed on GAL4 binding sites, regions YII and YIII were each sufficient to confer activation in S. cerevisiae. Regions of ZEBRA which affected activation in S. cerevisiae were also required in human B lymphocytes. The amino-terminal region of ZEBRA (aa 1 to 98) was required for activation both in S. cerevisiae and in human B cells; deletion of the carboxy-terminal 18 aa also significantly reduced activation in both cell types. Thus, the behavior of ZEBRA in human B cells and S. cerevisiae suggests that the protein contains universal activation motifs that interact with conserved components of the transcription machinery. However, certain deletion mutants of ZEBRA containing mutations in the N-terminal region exhibited discordant behaviors in S. cerevisiae and in B cells. For example, deletion of ZEBRA aa 26 to 51 impaired activation to a great extent in B cells but had little or no effect in S. cerevisiae. The discordant mutants may reflect interactions with a variable domain of a conserved component or unique interactions with specialized components of the basal transcription apparatus in different cells. PMID- 8230469 TI - The cell surface receptor is a major determinant restricting the host range of the B-lymphotropic papovavirus. AB - The B-lymphotropic papovavirus (LPV) productively infects only a subset of human B-lymphoma-derived cell lines while transfection of the viral genome yields infectious viral particles in a much wider variety of human hematopoietic cell lines. We have analyzed the contribution of a putative LPV receptor on the cell surface of B-cell lines in restricting the virus host range. In order to establish a quantitative virus binding assay for LPV, infectious virus particles were highly purified by metrizamide equilibrium density centrifugation and used as immunogens to raise seven mouse monoclonal antibodies specific for LPV VP1. Virus particle binding was quantitated in an indirect, nonradioactive assay with an LPV VP1-specific enzyme-linked immunosorbent assay. Binding of LPV particles to permissive human B-lymphoma cell line BJA-B occurred within minutes. Kinetics and capacity of binding were similar at 4 and 37 degrees C. A BJA-B cell was estimated to bind approximately 600 virus particles at conditions under which 50% of the administered virus was bound. The sialidase and trypsin sensitivities of the cellular virus binding moiety show that sialylated and proteinaceous components are necessary components of the LPV receptor on BJA-B cells. Despite a high binding capacity of BJA-B cells for simian virus 40, LPV binding was not significantly affected by a 20-fold excess of simian virus 40 particles, indicating that these related polyomaviruses do not bind to the same receptor on BJA-B cells. Reduction of LPV binding to sialidase-pretreated BJA-B cells was accompanied by a similar reduction of infection, indicating that virus binding may be a limiting factor in the LPV replicative cycle. The two highly LPV permissive human B-lymphoma cell lines BJA-B and Namalwa displayed high virus binding whereas low and nonpermissive hematopoietic cell lines showed reduced or undetectable virus binding. We conclude that the inability of LPV particles to productively infect the nonpermissive human hematopoietic cell lines analyzed is probably due to the absence or insufficient expression of a functional cell surface receptor. PMID- 8230470 TI - The herpes simplex virus type 1 regulatory protein ICP0 enhances virus replication during acute infection and reactivation from latency. AB - ICP0 is a potent activator of herpes simplex virus type 1 gene expression in transient assays and in productive infection. A role for ICP0 in reactivation from latency in vivo has also been suggested on the basis of the observation that viruses with mutations in both copies of the diploid gene for ICP0 reactivate less efficiently than wild-type virus. Because the ICP0 gene is contained entirely within the coding sequences for the latency-associated transcripts (LATs), ICP0 mutants also contain mutations in LAT coding sequences. This overlap raises the question of whether mutations in ICP0 or the LATs, which have also been implicated in reactivation, are responsible for the reduced reactivation frequencies characteristic of ICP0 mutants. Two approaches were taken to examine more definitively the role of ICP0 in the establishment and reactivation of latency. First, a series of ICP0 nonsense, insertion, and deletion mutant viruses that exhibit graded levels of ICP0-specific transactivating activity were tested for parameters of the establishment and reactivation of latency in a mouse ocular model. Although these mutants are ICP0 LAT double mutants, all nonsense mutants induced the synthesis of near-wild-type levels of the 2-kb LAT, demonstrating that the nonsense linker did not disrupt the synthesis of this LAT species. All mutants replicated less efficiently than the wild-type virus in mouse eyes and ganglia during the acute phase of infection. The replication efficiencies of the mutants at these sites corresponded well with the ICP0 transactivating activities of individual mutant peptides in transient expression assays. All mutants exhibited reduced reactivation frequencies relative to those of wild-type virus, and reactivation frequencies, like replication efficiencies in eyes and ganglia, correlated well with the level of ICP0 transactivating activity exhibited by individual mutant peptides. The amount of DNA of the different mutants varied in latently infected ganglia, as demonstrated by polymerase chain reaction analysis. No correlation was evident between reactivation frequencies and the levels of viral DNA in latently infected ganglia. Thus, replication and reactivation efficiencies of ICP0 mutant viruses correlated well with the transactivating efficiency of the corresponding mutant peptides. In a second approach to examining the role of ICP0 in latency, a single copy of the wild-type gene for ICP0 was inserted into the genome of an ICP0- LAT- double mutant, 7134, which exhibits a marked impairment in its ability to replicate in the mouse eye and reactivate from latency.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8230471 TI - Divergence and evolution of homologous regions of Bombyx mori nuclear polyhedrosis virus. AB - Homologous regions (hrs) (hr1,hr2-left,hr2-right,hr3,hr4-left,hr 4-right, and hr5) similar to those found in the Autographa californica nuclear polyhedrosis virus (AcNPV) genome were found in the Bombyx mori NPV (BmNPV) genome. The BmNPV hrs contained two to eight repeats of a homologous nucleotide sequence which were on average about 75 bp long. All of these homologous sequence repeats contained a 26-bp-long palindrome motif with an EcoRI or EcoRI-like site at its core. The consensus sequence of the BmNPV hrs showed 95% conservation with respect to those found in AcNPV. Nucleotide sequence analysis indicated that hr2-left and hr2 right of BmNPV evolved from an ancestor similar to hr2 of AcNPV by inversion, cleavage, and ligation. The polarities of the BmNPV and AcNPV hrs were conserved except for that of hr4-left. Within hr4-right of BmNPV, four repeats of a previously underscribed palindrome motif were found. Bmhr5D, a BmNPV mutant which lacked hr5, replicated at a rate similar to that of wild-type BmNPV in BmN cells and silkworm larvae, indicating that hr5 was not essential for viral replication. After ten passages of Bmhr5D in BmN cells, no detectable changes in its genome were observed by restriction endonuclease analysis. The evolution and divergence of the BmNPV genome are also discussed. PMID- 8230472 TI - Dynamic equilibrium between vesicular stomatitis virus glycoprotein monomers and trimers in the Golgi and at the cell surface. AB - Previous studies have shown that trimers of the vesicular stomatitis virus glycoprotein (VSV G protein) are in rapid equilibrium with monomeric subunits after folding and assembly in the endoplasmic reticulum (ER). To determine whether G protein trimers were in equilibrium with monomers in other cellular compartments, we studied heterotrimer formation between VSV G protein and a mutant G protein (G mu protein) containing a 3-amino-acid cytoplasmic domain replacing the normal 29-amino-acid domain. The G mu protein is transported from the ER much more slowly than G protein, although both G and G mu proteins form trimers rapidly in the ER. In coexpression experiments, we observed that VSV G protein molecules exited the ER about sixfold faster than G mu protein molecules, and we observed no heterotrimer formation in the ER, probably because of rapid reassortment of the mutant and wild-type trimers. However, heterotrimer formation between the two proteins was observed after long chase periods that allowed time for trimers of the mutant protein to reach the plasma membrane and reassort with the G protein subunits. Additional studies showed that heterotrimers of the two proteins could form in the Golgi or in the ER if exit of the G protein from either compartment was blocked. PMID- 8230473 TI - Human cytomegalovirus IE86 protein interacts with promoter-bound TATA-binding protein via a specific region distinct from the autorepression domain. AB - The major immediate-early gene of human cytomegalovirus encodes several isoforms of an immediate-early protein which has distinct transcriptional regulatory properties. The IE86 isoform autorepresses the major immediate-early promoter by directly binding the cis repression signal element located between the TATA box and the mRNA cap site. In addition to this activity, IE86 stimulates other viral and cellular promoters. One mechanism by which eukaryotic regulatory proteins are thought to stimulate transcription is by contacting one or more general transcription factors. We show that the IE86 protein physically interacts with the DNA-binding subunit (TATA-binding protein) human transcription factor IID via the TATA-binding protein-contacting domain in the N terminus of IE86. In a mobility shift assay, IE86 was also observed to stabilize the binding of TATA binding protein to promoter DNA. The domains within IE86 responsible for mediating transactivation and repression functioned independently. These experiments thus demonstrate the elegant ability of human cytomegalovirus to join different protein domains to produce distinct multifunctional proteins. PMID- 8230474 TI - Site-specific inhibition of RNA polymerase II preinitiation complex assembly by human cytomegalovirus IE86 protein. AB - The human cytomegalovirus major immediate-early gene encodes several protein isoforms which autoregulate the major immediate-early promoter (MIEP). One of these isoforms, the IE86 protein (UL122, IE2), is a DNA-binding protein that represses the MIEP through its cognate recognition sequence (designated the cis repression signal [crs]) located between the TATA box and the initiation site of transcription. Purified recombinant IE86 protein was shown to repress MIEP transcription in vitro, in a cis-acting mediated pathway, with nuclear extracts from HeLa S3, U373-MG, and primary human foreskin fibroblast cells. Repression of the MIEP by IE86 was shown by two criteria to be dependent on the direct interaction of IE86 with the crs element. Core promoter constructs containing essentially the MIEP TATA box and crs element were also specifically repressed by IE86 but not by a mutant IE86 protein, indicating the general transcription machinery as the target for IE86 repression. Kinetic and template commitment experiments demonstrated that IE86 affects preinitiation complex formation but not the rate of reinitiation. Sarkosyl inhibition experiments further revealed that IE86 was unable to effect repression by either disassembling or preventing the elongation of a preexisting transcription complex. Further, the ability of IE86 to interact with the DNA-binding subunit of TFIID was shown not to be required for repression. These functional protein-DNA and protein-protein interaction experiments demonstrate that IE86 specifically interferes with the assembly of RNA polymerase II preinitiation complexes. The biological significance of these results and the precise mechanism by which IE86 represses transcription are discussed. PMID- 8230475 TI - Adenovirus E4orf4 protein binds to protein phosphatase 2A, and the complex down regulates E1A-enhanced junB transcription. AB - Adenovirus E4orf4 protein was previously shown to counteract transactivation of junB by cyclic AMP (cAMP) and E1A protein. It was also shown to cause hypophosphorylation of E1A and c-Fos proteins. Here we show that the E4orf4 protein associates with protein phosphatase 2A. All three subunits of the phosphatase are present in the complex, and the B subunit interacts directly with the viral protein. The complex possesses a phosphatase activity typical of protein phosphatase 2A, and the phosphatase mediates the E4orf4-induced down regulation of junB transcription. Thus, adenovirus E4orf4 protein recruits protein phosphatase 2A into a signal transduction pathway initiated by cAMP and E1A protein. PMID- 8230476 TI - Vaccinia virus nucleoside triphosphate phosphohydrolase I controls early and late gene expression by regulating the rate of transcription. AB - We have carried out a detailed analysis of viral mRNAs and proteins produced in cultured cells infected with a temperature-sensitive vaccinia virus mutant (ts36) containing a modified nucleoside triphosphate phosphohydrolase I (NPH-I), a nucleic acid-dependent ATPase. Using a recombinant virus (ts36LUC) which expresses the luciferase marker, we showed in seven different cell lines that early expression of the receptor gene is strongly inhibited (73.8 to 98.7%) at the nonpermissive temperature. The steady-state levels of different early viral polypeptides were also severely reduced. Analysis of steady-state mRNA levels for two early genes (DNA polymerase and D5) showed that inhibition of early polypeptide synthesis correlated with a reduction in the levels of mRNA accumulated at the nonpermissive temperature. Analysis of steady-state levels of late viral polypeptides and of mRNAs indicated that NPH-I regulation of intermediate and late gene expression is direct and not simply a consequence of its role in inhibiting early gene expression. Characterization of a rescued virus (R36) demonstrated that the temperature-sensitive phenotype of ts36 is due solely to the point mutation in the NPH-I gene. The mutant phenotype is not due to reduced levels of NPH-I present in ts36 virions or to the differential stability of this enzyme in cells infected at the nonpermissive temperature but to inhibition of normal enzymatic activity for this protein. Measurement of viral transcriptional activity in permeabilized purified virions demonstrated that NPH I is required for normal rates of transcription in vaccinia virus. Our findings show ts36 to be a strongly defective early mutant of vaccinia virus and prove that NPH-I plays a key role in the control of early and late virus gene expression, possibly by way of an auxiliary function which regulates mRNA transcription during the virus growth cycle. PMID- 8230477 TI - Varicella-zoster virus DNA polymerase and major DNA-binding protein genes have overlapping divergent promoters. AB - A detailed analysis of the transcriptionally divergent promoters of varicella zoster virus (VZV) open reading frames (ORFs) 28 and 29, encoding the DNA polymerase and major DNA-binding proteins, respectively, was performed. We found that the 221-bp ORF 28-29 intergenic domain contains overlapping divergent promoters; these promoters have TATA boxes and cap sites arranged closely back-to back, have highly concordant patterns of responsiveness to transactivation by VZV ORFs 4 and/or 62, and could not be separated without abolishing the effects that VZV trans activators imparted to them. Mutation of the ORF 28 TATA box rendered this promoter unresponsive to ORF 62 and the combination of ORFs 4 and 62 without altering ORF 29 promoter activity. Mutations of all potential ORF 29 TATA boxes collectively failed to abolish this promoter's responsiveness to either ORF 4 or ORF 62, suggesting a mechanism of gene regulation for ORF 29 that differs from that of ORF 28. These findings are concordant with the observation that both genes are expressed in productive infection, but only ORF 29 expression has been identified in latency. PMID- 8230478 TI - Role of spike protein conformational changes in fusion of Semliki Forest virus. AB - The alphavirus Semliki Forest virus (SFV) and a number of other enveloped animal viruses infect cells via a membrane fusion reaction triggered by the low pH within endocytic vesicles. In addition to having a low pH requirement, SFV fusion and infection are also strictly dependent on the presence of cholesterol in the host cell membrane. A number of conformational changes in the SFV spike protein occur following low-pH treatment, including dissociation of the E1-E2 dimer, conformational changes in the E1 and E2 subunits, and oligomerization of E1 to a homotrimer. To allow the ordering of these events, we have compared the kinetics of these conformational changes with those of fusion, using pH treatment near the fusion threshold and low-temperature incubation to slow the fusion reaction. Dimer dissociation, the E1 conformational change, and E1 trimerization all occur prior to the mixing of virus and cell membranes. Studies of cells incubated at 20 degrees C showed that as with virus fusion, E1 trimerization occurred in the endosome before transport to lysosomes. However, unlike the strictly cholesterol dependent membrane fusion reaction, the E1 homotrimer was produced in vivo during virus uptake by cholesterol-depleted cells or in vitro by low-pH treatment of virus in the presence of artificial liposomes with or without cholesterol. Purified, lipid-free spike protein rosettes were assayed to determine the requirement for virus membrane cholesterol in E1 homotrimer formation. Spike protein rosettes were found to undergo E1 oligomerization upon exposure to low pH and target liposomes and showed an enhancement of oligomerization with cholesterol-containing membranes. The E1 homotrimer may represent a perfusion complex that requires cholesterol to carry out the final coalescence of the viral and target membranes. PMID- 8230479 TI - The DNA-binding domain of simian virus 40 tumor antigen has multiple functions. AB - The DNA-binding domain of simian virus 40 tumor antigen has been previously shown to participate in a number of different activities. Besides being involved in binding to sequences at the viral replication origin, this domain appears to be required for nonspecific DNA binding, for structurally distorting origin DNA (melting and untwisting), and possibly for oligomerization of the protein into hexamers and double hexamers. We now provide evidence that it also takes part in unwinding origin DNA sequences, contributes a function specifically related to in vivo DNA replication, and perhaps supports the assembly of the virus or release of the virus from the cell. This 100-amino-acid domain appears to be an excellent model system for studying how a small region of a protein could have a number of distinct activities. PMID- 8230480 TI - NF-kappa B p100 is one of the high-molecular-weight proteins complexed with the v Rel oncoprotein in transformed chicken spleen cells. AB - The Rel/NF-kappa B family of proteins includes several interacting cellular transcription factors and the v-Rel oncoprotein of the avian Rev-T retrovirus. We report the isolation of a chicken cDNA for the NF-kappa B p52 precursor protein p100. Full-length p100 only weakly binds DNA in vitro; removal of the ankyrin like repeats generates C-terminally truncated p100 proteins (like p52) that have an increased ability to bind an oligonucleotide containing a kappa B site. In addition, we show that chicken p100 is identical to a protein previously designated p115, which is found in a complex with v-Rel in transformed chicken spleen cells. Furthermore, p100 and v-Rel can form a complex when synthesized in vitro. Using cDNAs for chicken NF-kappa B p105, NF-kappa B p100, c-Rel, and v Rel, we show that one of the complexes in v-Rel-transformed spleen cells can be reconstituted in vitro. PMID- 8230481 TI - Effects of virally expressed interleukin-10 on vaccinia virus infection in mice. AB - To investigate the in vivo role of interleukin-10 (IL-10) in viral infection, we compared infections with a recombinant vaccinia virus (VV) expressing IL-10 (VV IL10) under control of the VV P7.5 promoter and a control virus (VV-beta gal) in normal and severe combined immunodeficient mice. In normal mice, VV-IL10 infection resulted in less natural killer cell activity at 3 days postinfection and less VV-specific cytotoxic T-cell activity at 6 or 7 days postinfection than VV-beta gal infection. However, the use of dermal scarification or intraperitoneal, intranasal, or intracerebral inoculation into immunocompetent mice resulted in no difference between VV-IL10 and VV-beta gal in visible lesions, mortality, protective immunity to a 100-fold lethal VV challenge, or VV specific antibody response. In the immunodeficient mice, VV-IL10 infection resulted in greater natural killer cell activity and lower virus replication than VV-beta gal infection. These in vivo effects were subtler and more complex than had been anticipated. From the VV-IL10 murine model, the Epstein-Barr virus encoded homolog of human IL-10, BCRF1, may provide a selective advantage by blunting the early human natural killer cell and cytotoxic T-cell responses so that Epstein-Barr virus can establish a well-contained latent infection in B lymphocytes. PMID- 8230482 TI - Mutants of Epstein-Barr virus with a selective marker disrupting the TP gene transform B cells and replicate normally in culture. AB - We have isolated mutants of Epstein-Barr virus (EBV) which carry a dominant selectable marker inserted into the third exon of the gene encoding two membrane proteins, TP1 and TP2 (or LMP2A and LMP2B), which are expressed in latently infected, growth-transformed B cells. One of the mutants also acquired a 260-bp deletion beginning in the first intron a few base pairs from the terminal repeats and removing most of the second TP exon, including the initial coding sequences of TP2. These EBV mutants transform human B cells in culture, and the transformed B-cell clones carrying them release EBV at approximately normal frequencies. PMID- 8230483 TI - Analysis of p107-associated proteins: p107 associates with a form of E2F that differs from pRB-associated E2F-1. AB - The binding of viral oncogenes to cellular proteins is thought to modulate the activities of these cellular targets. The p107 protein is targeted by many viral proteins, including adenovirus E1A, simian virus 40 large T antigen, and human papillomavirus type 16 E7 protein. A panel of monoclonal antibodies against p107 was raised and used to identify cellular proteins that interact with the p107 protein in vivo. p107-associated proteins included cyclin A, cyclin E, and cdk2. In addition, p107 was found to associate with 62- to 65- and 50-kDa phosphoproteins in ML-1 cells, a human myeloid leukemia cell line. The 62- to 65 kDa proteins have many of the properties of the transcription factor E2F but were distinguished from pRB-associated E2F-1 by both immunologic and biochemical properties. PMID- 8230484 TI - Functional interaction between Epstein-Barr virus DNA polymerase catalytic subunit and its accessory subunit in vitro. AB - The Epstein-Barr virus (EBV) DNA polymerase catalytic subunit (BALF5 protein) and its accessory subunit (BMRF1 protein) have been independently overexpressed and purified (T. Tsurumi, A. Kobayashi, K. Tamai, T. Daikoku, R. Kurachi, and Y. Nishiyama, J. Virol. 67:4651-4658, 1993; T. Tsurumi, J. Virol. 67:1681-1687, 1993). In an investigation of the molecular basis of protein-protein interactions between the subunits of the EBV DNA polymerase holoenzyme, we compared the DNA polymerase activity catalyzed by the BALF5 protein in the presence or absence of the BMRF1 polymerase accessory subunit in vitro. The DNA polymerase activity of the BALF5 polymerase catalytic subunit alone was sensitive to high ionic strength on an activated DNA template (80% inhibition at 100 mM ammonium sulfate). Addition of the polymerase accessory subunit to the reaction greatly enhanced DNA polymerase activity in the presence of high concentrations of ammonium sulfate (10-fold stimulation at 100 mM ammonium sulfate). Optimal stimulation was obtained when the molar ratio of BMRF1 protein to BALF5 protein was 2 or more. The DNA polymerase activity of the BALF5 protein along with the BMRF1 protein was neutralized by a monoclonal antibody to the BMRF1 protein, whereas that of the BALF5 protein alone was not, suggesting a specific interaction between the BALF5 protein and the BMRF1 protein in the reaction. The processivity of nucleotide polymerization of the BALF5 polymerase catalytic subunit on singly primed M13 single-stranded DNA circles was low (approximately 50 nucleotides). Addition of the BMRF1 polymerase accessory subunit resulted in a strikingly high processive mode of deoxynucleotide polymerization (> 7,200 nucleotides). These findings strongly suggest that the BMRF1 polymerase accessory subunit stabilizes interaction between the EBV DNA polymerase and primer template and functions as a sliding clamp at the growing 3'-OH end of the primer terminus to increase the processivity of polymerization. PMID- 8230485 TI - Epstein-Barr virus induces fragmentation of chromosomal DNA during lytic infection. AB - Pulsed-field agarose gel electrophoresis showed that fragmentation of chromosomal DNA in Raji cells was induced by infection with the P3HR-1 strain of Epstein-Barr virus (EBV). S1 nuclease treatment of the agarose plugs containing cells suggested that the majority of DNA fragments did not contain single-strand gaps. Chromosomal DNA fragmentation was inhibited by cycloheximide, indicating that protein synthesis was required for DNA fragmentation. Phosphonoacetic acid, an inhibitor of EBV DNA polymerase, did not inhibit fragmentation of chromosomal DNA. These findings suggest that EBV-specific early proteins participate in fragmentation of chromosomal DNA. Chromosomal DNA of P3HR-1 cells was also fragmented by treatment with n-butyrate plus 12-O-tetradecanoylphorbol-13-acetate (TPA), which induced activation of latent EBV genome following viral replication. In addition, fragmentation of DNA preceded cell death during lytic infection. These results suggest that fragmentation of chromosomal DNA is generally induced during EBV replication and probably contributes to the cytopathic effect of EBV. The role of DNA fragmentation in death of infected cells is discussed in relation to apoptosis. PMID- 8230486 TI - Isoprenylation mediates direct protein-protein interactions between hepatitis large delta antigen and hepatitis B virus surface antigen. AB - Hepatitis delta antigen (HDAg) consists of two protein species of 195 and 214 amino acids, respectively, which are identical in sequence except that the large HDAg has additional 19 amino acids at its C terminus and is prenylated. Previous studies have shown that the large HDAg and the surface antigen of hepatitis B virus (HBsAg) together can form empty hepatitis delta virus (HDV) particles. To understand the molecular mechanism of HDV virion morphogenesis, we investigated the possible direct protein-protein interaction between HDAg and HBsAg. We constructed recombinant baculoviruses expressing the major form of HBsAg and various mutant HDAgs and used these proteins for far-Western protein binding assays. We demonstrated that HBsAg interacted specifically with the large HDAg but not with the small HDAg. Using mutant HDAgs which have defective or aberrant prenylation, we showed that this interaction required isoprenylates on the cysteine residue of the C terminus of the large HDAg. Isoprenylation alone, without the remainder of the C-terminal amino acids of the large HDAg, was insufficient to mediate interaction with HBsAg. This study demonstrates a novel role of prenylates in HDV virion assembly. PMID- 8230487 TI - Identification and characterization of an extended Tax protein in human T-cell lymphotropic virus type II subtype b isolates. AB - The tax gene sequence of human T-cell lymphotropic virus type II isolate G12 (HTLV-IIG12) was found to encode an extended Tax protein when compared with that of HTLV-IIMoT. In vitro transcription-translation of the HTLV-IIG12 tax gene produced a 40-kDa Tax protein that specifically reacted with serum specimens from HTLV-II-infected individuals. Limited sequence analysis demonstrated that isolates with an extended Tax protein were all HTLV-II subtype b (HTLV-IIb). Therefore, the extended Tax protein appears to be a unique characteristic of most HTLV-IIb isolates and may be useful in designing immunoassays to distinguish between HTLV-IIa and HTLV-IIb. PMID- 8230488 TI - Interconnection between thyroid hormone signalling pathways and parvovirus cytotoxic functions. AB - Nonstructural (NS) proteins of autonomous parvoviruses can repress expression driven by heterologous promoters, an activity which thus far has not been separated from their cytotoxic effects. It is shown here that, in transient transfection assays, the NS-1 protein of the parvovirus minute virus of mice (MVMp) activates the promoter of the human c-erbA1 gene, encoding the thyroid hormone (T3) receptor alpha. The endogenous c-erbA1 promoter is also a target for induction upon MVMp infection. Moreover, T3 was found to up-modulate the level of cell sensitivity to parvovirus attack. These data suggest an interconnection between T3 signalling and NS cytotoxic pathways. PMID- 8230489 TI - Immunization with the immediate-early tegument protein (open reading frame 62) of varicella-zoster virus protects guinea pigs against virus challenge. AB - The IE62 protein, the primary regulatory protein of varicella-zoster virus (VZV) and the major component of the virion tegument, was an effective immunogen in the guinea pig model of VZV infection, whereas the ORF 29 gene product, a nonstructural DNA replication protein, did not elicit protection. All animals immunized with the ORF 29 protein had cell-associated viremia compared with 2 of 11 guinea pigs given the IE62 protein (P = 0.005). VZV was detected in ganglia from 38% of the animals given the ORF 29 protein and 44% of the control animals compared with 9% of the animals immunized with the IE62 protein (P = 0.04). In contrast to the IE62 protein, immunization with the ORF 29 protein did not prime the animals for an enhanced T-cell response upon challenge with infectious virus. The VZV IE62 protein has potential value as a vaccine component. PMID- 8230490 TI - Identification of a lytic-phase origin of DNA replication in human herpesvirus 6B strain Z29. AB - DNA sequences which have structural features suggestive of their functioning as an origin of lytic-phase DNA replication were previously identified in both human herpesvirus 6B strain Z29 [HHV-6B (Z29)] and in HHV-6A (U1102). Plasmid constructs containing the putative HHV-6B (Z29) oriLyt element were replicated after transfection into permissive T cells, when trans-acting factors were provided by HHV-6B (R-1) infection. By using this assay, the HHV-6B (Z29) oriLyt was mapped to a minimal region of approximately 400 bp which lies upstream of the gene that is homologous to herpes simplex virus UL29, a region that carries an origin in other betaherpesviruses and in some alphaherpesviruses. PMID- 8230491 TI - Multiple, distinct trans-activation functions are encoded by the simian virus 40 large T and small t antigens, only some of which require the 82-residue amino terminal common domain. AB - Simian virus 40 (SV40) small t and large T antigens can each trans activate the adenovirus (Ad) E2A and the Ad VA-I promoters. The first 82 amino acids of large T and small t are identical. However, this large T-small t common domain between residues 1 and 82 does not trans activate, suggesting that large T and small t each encode separate trans-activation functions. To determine whether the large T or small t unique domains, which are required for trans activation of the E2A promoter, are sufficient for this activity, we have employed expression plasmids separately encoding the common and unique domains of large T and small t. Cotransfection of a large T unique domain expression plasmid efficiently trans activated the E2A promoter. Optimal trans activation by large T required the motif that binds cellular proteins such as the retinoblastoma gene product, which is located in the large T unique domain, and additional large T structures outside this motif. In contrast, the small t unique domain did not trans activate the E2A promoter. Experiments utilizing E2A promoter mutants containing only the ATF- or EIIF-binding sites demonstrated that trans activation by small t involves only the EIIF transcription factor and that this function requires both the common (residues 1 to 82) and the small t unique domains expressed as a colinear protein. trans activation by large T, in contrast, involves at least three mechanisms. There appear to be at least two mechanisms that involve the EIIF transcription factor, at least one of which does not require the common domain (residues 1 to 82) and one mechanism that involves the ATF factor and does require both the common and the large T unique domains. PMID- 8230492 TI - Mouse mammary tumor virus-induced tumorigenesis in sag transgenic mice: a laboratory model of natural selection. AB - Transgenic mice that expressed the superantigen protein encoded in the C3H exogenous mouse mammary tumor virus long terminal repeat deleted their V beta 14+ T cells during the shaping of their immune repertoire and showed no evidence of virus production in their mammary glands after infection by milk-borne C3H exogenous virus. However, they developed mammary gland tumors that had newly integrated copies of C3H exogenous virus, although the latency of tumor formation was much longer than in their nontransgenic littermates that retained their V beta 14+ T cells. After four generations, infectious C3H virus was completely eliminated from the transgenic mouse pedigree. These data support the hypothesis that endogenous mouse mammary tumor proviruses are retained in the genome as protection against exogenous virus infection and subsequent tumorigenesis and show that there may be natural selection against the virus in vivo. PMID- 8230493 TI - The human T-cell leukemia/lymphotropic virus type I p12I protein cooperates with the E5 oncoprotein of bovine papillomavirus in cell transformation and binds the 16-kilodalton subunit of the vacuolar H+ ATPase. AB - The human T-cell leukemia/lymphotropic virus type I (HTLV-I) induces T-cell leukemia and transforms human T cells in vitro. A recently identified protein with a molecular weight of 12,000 (12K) (p12I), encoded by single- and double spliced mRNAs transcribed from the 3' end of the HTLV-I genome, has been shown to localize in the perinuclear compartment and in the cellular endomembranes. The p12I protein exhibits significant amino acid sequence similarity to the E5 oncoprotein of bovine papillomavirus type 1 (BPV-1). Both proteins are very hydrophobic, contain a glutamine residue in the middle of a potential transmembrane region(s), and are localized in similar cellular compartments. Because of these observations, we investigated whether the p12I resemblance to E5 correlated with a similarity in their biological behavior. We expressed the p12I protein to evaluate its ability to functionally cooperate with the BPV-1 E5 oncoprotein and to bind to a cellular target of the E5 protein, the 16K component of the vacuolar H+ ATPase. Cotransfection of the mouse C127 cell line with the p12I and E5 cDNAs showed that although p12I alone could not induce focus formation, it strongly potentiated the transforming activity of E5. In addition, the p12I protein bound to the 16K protein as efficiently as the E5 protein. These findings might provide new insight for potential mechanisms of HTLV-I transformation and suggest that p12I and E5 represent an example of convergent evolution between RNA and DNA viruses. PMID- 8230494 TI - Molecular mimicry between the human immunodeficiency virus type 1 gp120 V3 loop and human brain proteins. AB - Immunologically cross-reactive proteins in the human brain that resemble the V3 loop of human immunodeficiency virus type 1 (HIV-1) gp120 have been identified. When several homogenized tissues from normal brains were used, a monoclonal antibody raised against amino acids 308 to 320 of the V3 loop reacted with three prominent human brain proteins (HBP) of 35, 55, and 110 kDa. Among the three, the 55-kDa HBP appears to be specific to the central nervous system. These results indicate that the V3 loop of HIV-1 gp120 shares an epitope with HBP. An immune response to the V3 loop that generates cross-reactive antibodies to cellular proteins may be an autoimmune mechanism by which HIV-1 can damage the central nervous system. PMID- 8230495 TI - The management of clinically unconfirmed positive urinary cytology. AB - An aggressive evaluation of extravesical sites of disease in patients with clinically unconfirmed positive urinary cytology findings is indicated only in previously untreated asymptomatic or symptomatic patients (group 1) and in those with a complete response to intravesical therapy who are without evidence of disease for 1 year (group 4). Patients who have positive urinary cytology findings immediately after complete transurethral resection of bladder tumors or intravesical therapy will almost always have recurrent bladder transitional cell carcinoma and do not require aggressive extravesical evaluation initially. Transurethral resection prostate biopsy between the 5 and 7 o'clock positions along the entire length of the prostatic urethra is the technique of choice for detecting transitional cell carcinoma of the prostate, since other less invasive techniques frequently underestimate the extent of disease. In the absence of radiographic disease, bilateral ureteral lavage cytology is the technique of choice to identify the upper tract as the source of a positive urinary cytology result. Rigid and flexible ureteropyeloscopy should be performed only in the presence of radiographic or cytological evidence of upper tract transitional cell carcinoma, or in patients with a history of upper tract disease who have suspected relapse on the basis of a positive urinary cytology result. PMID- 8230496 TI - Hot occupation and nephrolithiasis. AB - We investigated the prevalence of stone disease and urinary stone risk factors in machinists chronically exposed to a hot environment and massive sweating, without interference of nephrotoxic metals or other lithogenic compounds. The study was performed at a glass plant and exposure to heat stress was estimated by the Wet Bulb Globe Temperature climatic index. The prevalence of nephrolithiasis on the entire population of the machinists was 8.5% (20 of 236), while the prevalence on the controls working in normal temperature was 2.4% (4 of 165) (p = 0.03). A high incidence (38.8%) of uric acid stones was present in the workers exposed to heat stress. Among the urinary stone risk indexes determined for 3 days during the 8 hour work shift on a randomly selected sample of 21 workers exposed and 21 workers not exposed to heat stress without any evidence of stone disease significant differences were found in uric acid concentration (722 +/- 195 versus 482 +/- 184 mg./l., p < 0.001), specific gravity (1,026 +/- 4 versus 1,021 +/- 6, p < 0.005) and pH (5.31 +/- 0.28 versus 5.64 +/- 0.54, p < 0.02), respectively. Thus, high uric acid relative supersaturation was present during occupation in hot temperatures (8.67 +/- 3.49) compared to occupation in normal temperatures (4.15 +/- 2.7) (p < 0.001). This study confirms that chronic dehydration represents a real lithogenic risk factor, mainly for uric acid stones, and adequate fluid intake is recommended during hot occupations. PMID- 8230497 TI - Randomized double-blind study of potassium citrate in idiopathic hypocitraturic calcium nephrolithiasis. AB - In an attempt to document the efficacy of potassium citrate in stone formation, 57 patients with active lithiasis (2 or more stones during the preceding 2 years) and hypocitraturia were randomly allocated into 2 groups, with 1 group taking 30 to 60 mEq. potassium citrate daily in wax matrix tablet formation and the other group receiving placebo. In 18 patients receiving potassium citrate for 3 years stone formation significantly declined after treatment from 1.2 +/- 0.6 to 0.1 +/ 0.2 per patient year (p < 0.0001), in 13 patients (72%) the disease was in remission and all patients showed a reduced stone formation rate individually. In contrast, 20 patients taking placebo medication for 3 years showed no significant change in stone formation rate (1.1 +/- 0.4 to 1.1 +/- 0.3 per patient year) and in only 4 patients (20%) was the disease in remission. The stone formation rate during potassium citrate treatment was significantly lower than during the placebo treatment (0.1 +/- 0.2 versus 1.1 +/- 0.3 per patient year, p < 0.001). Potassium citrate therapy caused a significant increase in urinary citrate, pH and potassium, whereas placebo did not. Adverse reactions to potassium citrate were mild causing only 2 patients in the potassium citrate group and 1 in the placebo group to withdraw from the study. In summary, our randomized trial showed the efficacy of potassium citrate in preventing new stone formation in idiopathic hypocitraturic calcium nephrolithiasis. PMID- 8230498 TI - Blood pressure changes after extracorporeal shock wave lithotripsy in normotensive patients. AB - To evaluate the blood pressure changes caused by extracorporeal shock wave lithotripsy 102 patients 5 to 81 years old (mean age 40 years) with normal blood pressure and kidney lithiasis were monitored during a mean period of 22 months. There were 61 male (group 1) and 41 female (group 2) patients. Patients were evaluated by measurement of the diastolic pressure and the average arterial pressure before and after lithotripsy. Hypertension was considered when the diastolic pressure was greater than 90 mm. Hg for 2 weeks. The amount of shock waves applied in each case ranged from 1,250 to 6,000, with a mean of 4,000 shock waves at a median intensity of 18.1 kv. The incidence of hypertension after extracorporeal shock wave lithotripsy was 3.92%, which is similar to that of a normal population, although the diastolic pressure was statistically higher after treatment in both groups. In the male patients the diastolic pressure increased from 79.26 (+/- 9.7) to 81.47 (+/- 10.1) mm. Hg and in female patients it ranged from 76.58 (+/- 8.3) to 79.26 (+/- 9.9) mm. Hg. Similarly, the average arterial pressure was equally higher in the female group, ranging from 89.88 to 91.75 mm. Hg. In the male group the difference was not statistically significant, despite an increase from 94.5 to 95.8 mm. Hg. PMID- 8230499 TI - Color flow Doppler sonography for extracorporeal shock wave lithotripsy. AB - This report documents the usefulness of color Doppler sonography for early verification of urinary stone fragmentation during extracorporeal shock wave lithotripsy with ultrasonographic focusing. In the experimental study lithotripsy was performed on human urinary stones placed in pig kidneys. Increasing color flow within the stone mass created by movement of small fragments indicated fragmentation earlier than pixel movement on the standard gray scale sonogram. The success of treatment was demonstrated macroscopically. In the clinical study 25 patients with radiopaque kidney stones were treated by color Doppler guided shock wave lithotripsy and compared to a control group of 32 patients with similar stone characteristics in whom the end point of treatment was chosen based on B-mode sonography alone. The amount of color flow within the stone reflex determined the end point of treatment. The degree of fragmentation was confirmed by fluoroscopy immediately after treatment. Disintegration was successful in all cases and fragments passed spontaneously. Color Doppler imaging during shock wave lithotripsy provides reliable information on the spatial and temporal characteristics of stone fragmentation, and is superior to B-mode sonography by providing more immediate, objective information on stone fragmentation and allowing better evaluation of the focal zone. With the help of this technique, the number of applied shock waves could be reduced by 20% compared with the control group. PMID- 8230500 TI - Urological evaluation and management of renal-proximity stab wounds. AB - The urological evaluation and management of renal-proximity stab wounds remain controversial. Questions regarding the "best" first radiographic study and the indications for nonoperative management are still unanswered. At the Los Angeles County-University of Southern California Medical Center we retrospectively reviewed the charts of 244 patients evaluated between January 1985 and December 1990 for renal-proximity stab wounds. There were 184 patients who presented without hematuria, 46 who presented with microscopic hematuria and 14 who presented with gross hematuria. When used as the first diagnostic study, an excretory urogram was 96% accurate in establishing the presence or absence of injury. In 34 patients additional evaluation was done with computerized tomography and/or angiography. A total of 43 injuries (17.6%) was found, of which 27 were successfully managed nonoperatively and 16 were managed surgically (1 nephrectomy). One patient had significant postoperative hemorrhage after partial nephrectomy, which was successfully managed with angiographic embolization. We conclude that although the degree of hematuria was a good indication of renal injury, the absence of hematuria did not preclude injury. In addition, excretory urography appears to be an acceptably reliable first study at our institution. We advocate a renal angiogram as the second radiographic study of choice because it reliably stages significant injuries and offers the possibility of therapeutic transcatheter embolization. Most renal injuries, when accurately staged, can safely be managed nonoperatively. PMID- 8230501 TI - Major renal lacerations with a devitalized fragment following blunt abdominal trauma: a comparison between nonoperative (expectant) versus surgical management. AB - Of 43 patients who sustained blunt trauma resulting in a major renal laceration with a devitalized fragment 27 had coexisting intraperitoneal and renal injuries for which emergency celiotomy and repair of the nonurological trauma were done. Management of the renal injuries consisted of simultaneous renal exploration with 23% urological morbidity in 13 cases or expectant management with 85% urological morbidity in 14 cases. In this latter patient population infected urinomas and perinephric abscesses seeded from coexisting enteric or pancreatic injuries were the most common complication (57%). The remaining 16 of the 43 patients sustained renal lacerations without associated intraperitoneal injuries and all were managed expectantly with 38% urological morbidity. These findings suggest that renal exploration and surgical repair significantly improve the prognosis only in patients with simultaneous intraperitoneal and renal injuries (p < 0.01). PMID- 8230502 TI - Renal trauma. PMID- 8230503 TI - Preoperative hospitalization to hydrate living kidney donors can be omitted without sacrificing graft function. AB - This prospective study was done to determine whether preoperative hospitalization for hydration of living kidney donors could be omitted without sacrificing graft function in the recipient. The study group consisted of 36 consecutive kidney donors who underwent donation on a "to come in" basis. Fluid management consisted of 1 l. lactated ringer's solution per hour beginning 2 hours preoperatively. The control group was hospitalized preoperatively for intravenous hydration of an equivalent amount of saline solution given for 12 to 16 hours. No recipient of the study group donors experienced delayed graft function. Furthermore, in that group immediate and longer term graft function, as defined by immediate graft output, serum creatinine on postoperative day 1 and nadir serum creatinine levels, was equivalent to or better than that in the control group. We conclude that a rapid infusion of saline solution in the immediate preoperative period can replace overnight intravenous hydration of living kidney donors, and that the result will be a significant reduction in overall hospital stay for the donor without compromise in graft function for the recipient. PMID- 8230504 TI - The natural history of renal angiomyolipoma. AB - Of 35 patients with 48 angiomyolipomas 24 patients were followed clinically to determine the natural history of angiomyolipoma. Average patient age at presentation was 50 years (range 17 to 74) and of the patients 94% were women, 17% had tuberous sclerosis and 25% overall had bilateral disease. The patients could be divided into 2 distinct groups based on tumor size of 4 cm. or less and greater than 4 cm. Those with tumors less than 4 cm. were less likely to be symptomatic (24%) and patients with angiomyolipomas greater than 4 cm. were more often symptomatic (52%). No surgery was required for tumors less than 4 cm. but for 30% of the tumors greater than 4 cm. surgical intervention was necessary. Unlike any previously reported large series this study included radiological and historical followup available for 24 patients with angiomyolipoma with a mean followup time of 4 years (range 0.5 to 14). Moreover, to our knowledge we report for the first time documented growth during the study period of 27% of angiomyolipomas less than 4 cm. (4 of 15 tumors) and 46% of angiomyolipomas greater than 4 cm. (6 of 13 tumors). All patients with tumors less than 4 cm. were asymptomatic and only 1 required surgery. In contrast, tumors greater than 4 cm. were more frequently symptomatic (46%) and required surgery (54%). Patients with tuberous sclerosis and angiomyolipomas were distinctly different from patients with angiomyolipoma only, since they tended to present at a younger age, had a higher incidence of bilateral renal involvement, were more symptomatic, had larger tumors that were more likely to grow, and frequently required surgery. Based on this study, a modified approach to the current management of angiomyolipoma is recommended. PMID- 8230505 TI - Ureteropelvic junction stenosis: vascular anatomical background for endopyelotomy. AB - To help endourologists perform endopyelotomy safely and efficiently with a reduced risk of vascular complications, we analyzed the vascular relationships to the ureteropelvic junction in 146, 3-dimensional endocasts of the kidney collecting system together with the intrarenal arteries and veins. There was a close relationship between a prominent vessel (artery and/or vein) and the anterior surface of the ureteropelvic junction in 65.1% of the cases, including the inferior segmental artery with a tributary of the renal vein in 45.2% and an artery or vein in 19.9%. In the remaining 34.9% of the cases the anterior surface of the ureteropelvic junction was free of vessels. There was a direct relationship between a prominent vessel (artery and/or vein) and the posterior surface of the ureteropelvic junction in 6.2% of the cases, including an artery and vein in 2.1%, and just an artery in 1.4%. In all cases (3.5%) of an artery crossing at the posterior surface of the ureteropelvic junction, this vessel was the posterior segmental artery (retropelvic artery). In 2.7% of the cases the relationship of the prominent vessel was just with a posterior tributary of the renal vein, and in 20.5% a vessel crossed lower than 1.5 cm. above the posterior surface of the ureteropelvic junction. Among these latter cases the vessel was an artery (posterior segmental artery) in 6.8%. In the remaining 73.3% of the cases the posterior surface was free of vessels up to 1.5 cm. above the ureteropelvic junction. Due to the anatomical findings, we advise that posterior and posterolateral incisions at the ureteropelvic junction be avoided, and that deep incision alongside the ureteropelvic junction stenotic wall be done only laterally. PMID- 8230506 TI - Organ entrapment and renal morcellation: permeability studies. AB - We collected 24 intact LapSacs* during a laparoscopy course at our institution. Following laparoscopic nephrectomy or nephroureterectomy in a pig using high speed electrical tissue morcellation, the sacks were examined for perforation. When filled with water, 4 of the 24 LapSacs (16%) were found to have small perforations. The 20 remaining sacks were randomly divided into 3 groups of 6 and 2 sacks, although intact, were kept as substitutes to use if a trial with the study sacks could not be completed. Each group of 6 sacks was tested for permeability using 10 mg./ml. bovine serum albumin, 8 mg./ml. indigo carmine or 1 x 10(5)/ml. mouse bladder tumor cells. The results in the 18 tested sacks showed no dialysis of bovine serum albumin in the dialysate at 1, 3 and 18 hours, and no dialysis of indigo carmine at 0, 1, 2 and 3 hours. In 1 of 6 sacks tested with a mouse bladder tumor cell line a single cell was noted on the hemocytometer at 1 and 3 hours. The lack of an increasing concentration of cells at the 3-hour interval likely indicates that this represented contamination of the dialysate rather than ongoing leakage from a perforated sack in this case. We conclude that the LapSac, when properly used, remains impermeable throughout the morcellation process. PMID- 8230507 TI - Laparoscopic dismembered pyeloplasty. AB - As laparoscopic nephrectomy has become a viable ablative procedure for kidney removal, additional areas of reconstructive laparoscopic urological procedures are being investigated. We describe our early experience with laparoscopic pyeloplasty for the management of ureteropelvic junction obstruction. Technical highlights include initial placement of an internal ureteral stent, lateral insufflation, placement of 5, 10 mm. trocars, pyelotomy (or reduction pyeloplasty performed with articulating laparoscopic scissors, reapproximation of the ureteropelvic junction with a running 4-zero polyglactin suture, placement of a 7 mm. suction drain in the retroperitoneal space and reapproximation of the colon to the body wall with a hernia stapler. We have performed laparoscopic dismembered pyeloplasty in 5 patients with symptomatic ureteropelvic junction obstruction. Operating time ranged from 3 to 7 hours, with the majority of time devoted to laparoscopic suturing (1 to 3 hours). Hospital stay averaged 3 days and all patients returned to normal activity within 1 week. Followup averaged 12 months (range 9 to 17 months) with complete resolution of symptoms in all patients. We believe that this innovative reconstructive laparoscopic procedure can be used for treatment of complicated ureteropelvic junction obstruction as in patients with a large, redundant renal pelvis or crossing lower pole vessels. PMID- 8230508 TI - Stone granuloma: a cause of ureteral stricture. AB - Ureteral stricture is a recognized complication of ureteroscopy and ureteral stone fragmentation. Although most strictures are either asymptomatic or easily dilated, there are some strictures that result in progressive ureteral obstruction, do not respond to ureteral dilation and require operative intervention. A review of 125 percutaneous nephrostolithotomies for staghorn stone disease and 652 ureteroscopic stone fragmentations revealed 5 cases in which refractory ureteral strictures developed, requiring operative intervention. In 4 patients a "stone granuloma," embedded particles of calcium oxalate associated with macrophages and foreign body giant cells, was found with surrounding fibrosis and ureteral obstruction. In the remaining patient a suture granuloma from a recent ureterolithotomy was the source of the stricture. In each instance of stone granuloma the particles of calcium oxalate had become embedded in the wall as a consequence of ureteroscopic stone fragmentation and partial ureteral wall disruption. During ureteroscopy and intracorporeal lithotripsy every effort should be made to prevent calcium oxalate particles from becoming embedded in the ureteral wall. They are not inert and may cause irreversible stricture formation. To our knowledge, stone granuloma is a previously undescribed phenomenon and should be suspected when ureteral strictures that occur following ureteroscopy do not respond to endourological methods of management. PMID- 8230509 TI - Comparison of 2 pulsed lasers for lithotripsy of ureteral calculi: report on 154 patients. AB - Extracorporeal lithotripsy does not always provide satisfactory results for the treatment of ureteral stones. Such cases appear to be excellent indications for endocorporeal lithotripsy based on an association of ureteroscopy and laser. To compare the performances of 2 pulsed lasers, the pulsed dye laser (Candela) and solid Q switched laser (HMT), for the treatment of these calculi 161 ureteral stones were treated successively from November 1990 to March 1992 by a combination of ureteroscopy and laser. Endocorporeal lithotripsy was performed in 102 cases with the Candela laser, in 47 with the HMT laser and in 7 with both lasers. With a stable success rate greater than 90%, both lasers demonstrated equivalent performances regardless of the location of the stone along the ureter. However, while stone fragmentation was more rapid with the Candela laser, the HMT laser appeared to be more effective for dark stones (monohydrate calcium oxidate). PMID- 8230510 TI - Microstructural disorders of tunica albuginea in patients affected by Peyronie's disease with or without erection dysfunction. AB - Peyronie's disease, the etiology of which remains unknown, is characterized by a focal or multifocal disorder of penile tunica albuginea. An excessive collagen deposition gives rise to the formation of the plaque, which is fibrotic first and then becomes calcific. Typical symptoms of the disease are painful erection, pathological bending of the erect penis and impotence at the more advanced stages (43% of the cases). There is evidence that the tunica albuginea has a fundamental role in the erection mechanism. We evaluated the tunica albuginea of patients suffering from Peyronie's disease, with or without impotence, and found progressive disorders of the microstructure in areas that were not affected by fibrocalcific formations. There was a significant decrease (p < 0.0001) of elastic fiber concentration (55.05 +/- 23.83 per high power field) in patients with Peyronie's disease compared to the concentration in the tunica albuginea of a control group (128.50 +/- 11.79 per high power field). Moreover, when the elastic fiber concentration in the tunica albuginea of patients Peyronie's disease and normal erection (71.36 +/- 12.00 per high power field) was compared to the concentration in the tunica albuginea of those with Peyronie's disease and impotence (33.14 +/- 4.84 per high power field) a significant difference was noted (p < 0.0001). PMID- 8230511 TI - Dorsal vein sclerosis as a predictor of outcome in penile venous ligation surgery. AB - We studied the pathological specimens obtained from 24 impotent men who underwent proximal penile vein ligation and 6 potent men who underwent total penectomy as part of male-to-female transsexual surgery to determine if a correlation exists between venous pathology and surgical outcome. Pathology specimens, consisting of cavernosal tissue and penile veins, were independently reviewed by a single uropathologist (V. A.). The degree of vein wall thickness was carefully quantified for each patient. Venous leakage was documented by cavernosometry and cavernosography. Followup ranged from 7 to 19 months (mean 11 months). Within 6 months of the procedure 10 patients (41%) achieved rigid erections while 14 (59%) did not. Although no preoperative index could predict operative success, in the 10 patients with successful outcome histological examination of the excised vein segments revealed normal venous architecture with minimal vein wall thickness (calculated mean per cent vein wall thickness 39.4, range 17.5 to 51.7). In contrast, in the 14 patients in whom ligation failed there was marked vein wall thickness and sclerosis (calculated mean per cent vein wall thickness 68.0, range 55.3 to 85.6). In comparison, the vein segments obtained from the 6 potent patients had a wall thickness of 32.8% (range 8 to 39), which was equivalent to the 10 patients with postoperative erections. These findings suggest that there may be a correlation between vein wall thickness and the prognosis of patients who undergo venous ligation surgery for erectile dysfunction secondary to presumed corporeal venous incompetence. PMID- 8230512 TI - Early experience with the first pre-connected 3-piece inflatable penile prosthesis: the Mentor Alpha-1. AB - The aim of this clinical investigation was to obtain preliminary treatment outcome data concerning the Mentor Alpha-1, the first 3-piece inflatable penile prosthesis with pre-connected tubing between the pump and the penile cylinders. The study was designed to be the first multi-institutional treatment outcome report for any clinically available penile prosthesis with data analysis to be independent of the participating surgeons. A total of 12 board-certified urologists of mixed surgical training backgrounds and practices implanted the Alpha-1 device in 112 consecutive patients. With a mean of 27 +/- 5 months of followup the surgical complication rate included a 4% mechanical malfunction, 2% infection rate and 9% reoperation rate. Patient experience with the implanted device was computed from information on 96 of the 112 patients who returned a questionnaire. Of the patients 82% stated that the device fulfilled expectations as a treatment for impotence and 83% had improved sexual intercourse by 8 weeks after implantation. Patient satisfaction was computed on a scale of 12 equally weighted interrelated variables. Of the patients 77% recorded 9 or more cumulative satisfaction points. Patient and physician questionnaire data were analyzed for their relation to the cumulative prosthesis satisfaction score. A significant difference in cumulative scores was found for physician reported long term postoperative problems (mean satisfaction score 8.1 for patients with problems versus 10.2 for patients without problems, p = 0.018). The Alpha-1, with its feature of pre-connected tubing, is a reliable 3-piece inflatable penile prosthesis associated with a high level of patient satisfaction. PMID- 8230513 TI - Patient-partner satisfaction with semirigid penile prostheses for Peyronie's disease: a 5-year followup study. AB - The long-term acceptance of and satisfaction with penile prostheses were assessed in patients with Peyronie's disease and their partners. From 1985 to 1987, 50 men with advanced Peyronie's disease and associated erectile dysfunction were treated with a semirigid penile implant without additional plaque surgery. A total of 48 patients and 29 partners was reassessed at a followup of at least 60 months. Only 23 patients (48%) and 12 partners (40%) were totally satisfied with the long-term functional result and would repeat the same operation. Among the 25 dissatisfied patients (52%) loss of complementary erection resulting in a pencil-like penis, decrease in penile sensitivity, poor concealment and persistence of penile deviation were the major complaints. Of the partners 17 (60%) were dissatisfied due to poor penile girth, sensation of a cold glans penis, sensation of unnatural intercourse and dyspareunia. Eight patients (16%) chose to substitute the semirigid implant with a 3-component inflatable prosthesis. On a long-term basis, placement of a semirigid penile prosthesis for Peyronie's disease is associated with a significant patient-partner dissatisfaction rate. PMID- 8230514 TI - Experience with triple-drug therapy in a pharmacological erection program. AB - A group of 170 impotent men achieved usable erections during a 26-month period with a combination of papaverine, phentolamine and prostaglandin E1 (triple-drug therapy) injected intracorporeally. Of the patients 146 elected to enter a pharmacological erection program using this combination. Patient age ranged from 24 to 85 years and the average duration on the program was 11.2 months. Average injection volume was 0.36 cc per injection (range 0.1 to 1). Among those patients managed by our nurse clinician, only 3 episodes of priapism were encountered (1.7%). Scarring was documented in 7 of 170 patients (4.2%) 1 week to 21 months after starting the injections. Pain was encountered in 6 of 170 patients (3.5%). A superior dose response coupled with a low incidence of priapism, pain and scarring have led us to use triple-drug therapy as our agent of choice in the pharmacological management of erectile dysfunction. PMID- 8230515 TI - A prostaglandin E1 dose-response study in man. AB - Because prostaglandin E1 causes erection by smooth muscle relaxation in a receptor-dependent manner, one would expect increasing dosages to cause a progressively greater response and that, at receptor saturation, further increases would not be beneficial. To test this hypothesis a single-blind, placebo-controlled study of increasing dosages of prostaglandin E1 injected intracavernously was done. In 16 men with vasculogenic impotence erections were monitored by the RigiScan device in real time for 2 hours after injection, and rigidity, tumescence and duration of erection were measured. Summary parameters to characterize erection with each dosage were developed: maximal rigidity, maximal rigidity sustained for 30 minutes and duration of greater than 60% rigidity. The dose-response curve was similar for all 3 parameters. The initial response to escalating doses of prostaglandin E1 from 2.5 to 20 micrograms. demonstrated a steep dose-dependent increase; at greater than 20 micrograms. a plateau was reached, indicating a nonlinear response. More than 80% of the patients attained the maximal response at doses of 20 micrograms. or less and less than 20% benefited from a further increase. Based on these results, the effects of prostaglandin E1 appear to be receptor-dependent and prostaglandin E1 monotherapy for impotence could be limited to 20 micrograms. or less, since larger amounts offer little additional benefit. PMID- 8230516 TI - Treatment of erectile dysfunction. PMID- 8230517 TI - Mitomycin C versus estramustine in the treatment of hormone resistant metastatic prostate cancer: the final analysis of the European Organization for Research and Treatment of Cancer, genitourinary group prospective randomized phase III study (30865). AB - A total of 171 patients with progressive metastatic prostate cancer following hormonal therapy was randomized to receive either 560 to 700 mg. estramustine orally per day or 15 mg./m.2 mitomycin C by intravenous infusion every 6 weeks. The patients were recruited during a 2.5-year period, and 70% had undergone more than 1 previous therapy for prostate cancer, with some having received as many as 5 different previous treatments. The overall results were disappointing. The median time to progression was 5 months and 50% of the patients died within 10 months. There was no difference in efficacy between the 2 treatment arms. Toxicity was severe in both arms but appeared earlier in those patients receiving estramustine, leading to a tendency for earlier deterioration in performance status. In this group of heavily pretreated patients there appears to be no justification for the use of either of these agents at the present time. PMID- 8230518 TI - Treatment of clinical local failure after radiation therapy for prostate carcinoma. AB - From a base population of 634 patients with prostate cancer treated by external beam therapy with a median followup of 8 years and 123 patients treated by interstitial brachytherapy with 125iodine (125I) isotope with a median followup of 13 years, those with local failure only were identified. There were 57 external beam radiotherapy (9%) and 15 125I (12%) treated patients with local failure only among the base population. All but 3 patients (2 given external beam radiotherapy and 1 given 125I) were treated with hormonal manipulation without extirpative surgery. The overall cancer-specific median survival with hormonal therapy from the date of local failure was 70 months for 55 patients treated by external beam radiotherapy and 87 months for 14 treated by 125I. Patients with low grade, small volume tumors most likely to benefit from salvage surgery are also those who will experience prolonged survival with hormonal therapy. Patients with local failure only treated by hormonal manipulation had statistically longer cancer-specific survival rates from the date of failure than did similarly treated patients experiencing distant failure with local failure. This finding suggests a difference in the biological aggressiveness between tumors associated with distant and local failure versus local failure only. To select the patients with local failure only who would be candidates for the potentially benefited by salvage surgery, those with pretreatment stage A or B disease who were less than 72 years old were identified. A total of 17 patients treated by external beam radiotherapy and 7 treated by 125I fulfilled these criteria. Therefore, as determined by local failure only, patient age and pre-radiation clinical stage, only 2 to 5% of the patients treated with radiation modalities are ultimately optimal candidates for salvage surgery. PMID- 8230519 TI - Impact of transurethral resection on the long-term outcome of patients with prostatic carcinoma. AB - Between March 1970 and December 1987, 1,078 patients with adenocarcinoma of the prostate were treated with pelvic lymph node dissection and permanent 125iodine implantation. Before implantation, 257 patients (27%) underwent transurethral resection of the prostate, while 702 (73%) did not and their diagnosis was established by needle biopsy. A total of 119 patients (10%) underwent hormonal therapy before implantation and they were excluded from the present analysis. Clinical stage and pathological grade were similar in both groups. A higher percentage of patients in the transurethral resection group had nodal metastases at implantation. Positive lymph nodes were found in 121 patients (47%) in the transurethral resection group versus 199 (26%) who did not undergo resection (p < 0.001). The actuarial 5, 10 and 15-year distant metastasis-free survival rates among the patients who underwent transurethral resection of the prostate were 79%, 42% and 16%, respectively, compared to 86%, 52% and 27%, respectively, in the group without transurethral resection (p < 0.0001). Similarly, the actuarial disease-free and local relapse-free survival rates were significantly inferior in the transurethral resection group. A negative impact of transurethral resection of the prostate could be demonstrated among patients with grade I/II tumors. However, when stratified for nodal status, no difference in outcome in any clinical parameter was noted between the groups with and without transurethral resection of the prostate. Specifically, distant metastasis-free survival among transurethral resection group patients with negative nodes was 78%, 57% and 47% at 5, 10 and 15 years, respectively, compared to 80%, 59% and 47%, respectively, among the patients with negative nodes who did not undergo transurethral resection of the prostate (p = 0.38). Similarly, the differences between the 2 groups among patients with positive lymph nodes were not significant. When stratified by the clinical stage, grade and nodal status, the negative impact of transurethral resection of the prostate could not be demonstrated in any combination. A multivariate analysis failed to demonstrate transurethral resection of the prostate to be an independent variable in predicting the metastatic, local control or disease-free survival outcome. In conclusion, the long-term results in these pathologically staged cases indicate that transurethral resection of the prostate does not impact negatively on the clinical outcome. PMID- 8230520 TI - Prenatal adrenal hemorrhage confirmed by postnatal surgery. AB - A case is reported of massive adrenal hemorrhage in a newborn with evidence of bleeding in utero. A large cystic mass noted on prenatal ultrasonography at 20 weeks of gestation was followed by serial examinations until birth at 38 weeks of gestation. Postnatal evaluation included abdominal sonography, diethylenetriaminepentaacetic acid renal scan, voiding cystourethrography and abdominal plain film. Surgical exploration confirmed massive adrenal hemorrhage. PMID- 8230521 TI - Report of the Multicystic Kidney Registry: preliminary findings. AB - Multicystic kidney was first described in a 7-month-old child. Some physicians advocate nephrectomy in such cases, while others have questioned this procedure. In 1986 the American Academy of Pediatrics, Section on Urology established the Multicystic Kidney Registry to identify and follow patients with multicystic kidney disease. To date 441 patients have been registered, of whom 260 were managed nonoperatively. Based on followup of these patients no kidneys were removed for hypertension, infection, pain or tumor development. Long-term findings show that multicystic kidney disease may take as long as 20 years to resolve, and followup testing will be necessary. PMID- 8230522 TI - The multicystic dysplastic kidney and contralateral vesicoureteral reflux: protection of the solitary kidney. AB - The diagnosis of multicystic renal dysplasia is confirmed by a combination of sonography and radionuclide scan. If the contralateral kidney is normal by these criteria, further radiological examination is often omitted. We prospectively studied 29 patients with a diagnosis of multicystic renal dysplasia by voiding cystourethrography to determine the condition of the solitary contralateral kidney. Eight patients (28%) were found to have contralateral vesicoureteral reflux, of whom 6 had normal ultrasound examinations and would not have undergone further radiographic study based on ultrasound criteria alone. Contralateral vesicoureteral reflux in association with multicystic renal dysplasia puts solitary kidneys at risk for pyelonephritic scarring. Therefore, voiding cystourethrography should be performed as part of the initial evaluation of all infants with multicystic renal dysplasia. PMID- 8230523 TI - The Mitrofanoff principle in urinary reconstruction. AB - We report the use of the Mitrofanoff principle to achieve urinary continence in 47 patients. A catheterizable channel can be used as the primary continence mechanism, or as an adjunct to ensure complete bladder emptying if urethral catheterization or voiding is inadequate. Appendico-vesicostomy was performed in 25 of 47 patients (55%). Satisfactory continence was achieved in 45 patients (96%). Bladder emptying by clean intermittent catheterization was performed in all patients. Early complications included peristomal abscess (1 patient) and small bowel obstruction (1). Late complications included difficulty with catheterization (5 patients), stomal stenosis (9) and persistent incontinence (2). Our results support use of the appendix as the catheterizable segment of choice. We describe the technique and discuss the management of complications. PMID- 8230524 TI - The neurosurgical implications of continuous neurourological surveillance of children with myelodysplasia. AB - Between 1979 and 1990, 148 newborns with myelodysplasia were followed with serial urodynamic studies and neurological assessment. Of the patients 59 (40%) exhibited changes in neurological status by age 5 years, of whom 28 (19%) showed signs of deterioration. Most changes occurred before age 2 years. A total of 22 children underwent repeat neurosurgical exploration because of a change in urethral sphincter innervation (17), deterioration of function of the lower extremities (3), or changes on computerized tomography or magnetic resonance imaging (2). Postoperative urodynamic evaluation demonstrated improvement in 11 children, stabilization in 9 and further deterioration in 2. The earlier that a change was detected and secondary surgery was performed, the better the outcome. These findings indicate that the neurological lesion in myelodysplasia is a dynamic disease process requiring continuous neurological, orthopedic and urodynamic surveillance. Early identification and prompt neurosurgical reexploration seem to arrest and even reverse the neurological deterioration that takes place in a substantial number of these children. PMID- 8230525 TI - Isograft bladder mucosal transplantation for hypospadias repair in identical twins. AB - Urethral reconstruction with a bladder mucosal free graft offers several advantages for patients with complex hypospadias. We report a case in which identical twins circumcised at birth presented with mid-shaft hypospadias and chordee. A bladder mucosa graft was harvested from 1 twin, divided and used to form 2 neourethras. The donor twin underwent hypospadias repair, followed immediately by repair in the second twin with the isograft neourethra. To our knowledge this is the first reported case of bladder mucosal transplantation for hypospadias repair. PMID- 8230526 TI - Benign fibrous testicular pseudotumors in children. AB - Benign fibrous pseudotumors of the testicular tunics are rare in children. We report 2 cases of benign fibrous pseudotumors that were managed with gonad sparing surgery. We discuss the etiology, evaluation and treatment of benign fibrous pseudotumors of the testis. PMID- 8230527 TI - Ureteropelvic junction obstruction with renal pelvic calcification: a case report. AB - Calcification in the wall of the renal pelvis is rare. We report on a 65-year-old man with hydronephrosis secondary to ureteropelvic junction obstruction with renal pelvic calcification. Calcium deposit was found in the wall of the severely dilated renal pelvis. Pathological examination revealed a damaged and hyalinized fibrous renal pelvic wall and serum calcium level was normal. Thus, we speculated that this calcification was dystrophic. Chronic extensive dilatation with intermittent hemorrhage of the renal pelvic wall may have caused this dystrophic renal pelvic calcification. PMID- 8230528 TI - Laparoscopic pyeloplasty. AB - A 24-year-old woman presented with symptomatic right ureteropelvic junction obstruction due to anomalous lower pole renal vessels. A dismembered pyeloplasty of the ureteropelvic junction was performed completely laparoscopically. PMID- 8230529 TI - Chylothorax as a complication of radical nephrectomy. AB - We report a case of chylo-retroperitoneum and chylothorax following an uneventful left radical nephrectomy for renal cell carcinoma, and propose the mechanism of an infradiaphragmatic lymphatic injury with fistulous connection into the chest. Treatment with oral medium chain triglycerides was unsuccessful. Subsequently, total parenteral nutrition in conjunction with chest drainage, retroperitoneal drainage and sclerotherapy successfully treated the chylothorax. To our knowledge chylothorax following radical nephrectomy has not been reported previously in the literature. PMID- 8230530 TI - Breakage of a silicone double pigtail ureteral stent as a long-term complication. AB - A variety of indications have made the use of double pigtail ureteral catheters routine in urological practice. Although side effects are frequent they are usually mild. Complications with long-term indwelling stents are mainly due to stent incrustation or stone formation. We report a case of stent breakage without incrustation after an indwelling period of 29 months. To our knowledge such a severe complication of a silicone double pigtail stent has not been described previously. This complication also signifies that the period a stent is left indwelling is critical and should be monitored carefully. PMID- 8230531 TI - Uretero-fallopian fistula: an unusual complication of open ureterolithotomy. AB - We report a case of uretero-fallopian fistula, manifesting as complete urinary incontinence, following open ureterolithotomy for a lower ureteral calculus. PMID- 8230532 TI - Neurological manifestations of baclofen withdrawal. AB - Baclofen is a central nervous system agent that is commonly used for the treatment of muscle spasticity in spinal cord injury patients. Acute withdrawal of this medication can induce the development of neurological symptoms, including seizure disorder, psychosis, hallucinations and visual disturbances. We report 3 cases of acute central nervous system symptoms that developed in spinal cord injury patients. Each patient had been chronically maintained on a baclofen regimen to control muscle spasticity. Symptoms developed shortly after baclofen therapy was interrupted following genitourinary surgery. It is important that urologists become familiar with the symptomatology of baclofen withdrawal, the methods of its prevention and the appropriate therapy should the syndrome develop. PMID- 8230533 TI - Obstructive uropathy secondary to massive inguinoscrotal bladder herniation. AB - The bladder is partially involved in 1 to 4% of all inguinal hernias. However, few cases of massive inguinoscrotal herniation of the bladder have been described, all of which have involved unilateral hydronephrosis. We report on a 45-year-old man with incomplete voiding and massively enlarged right scrotum along with elevated creatinine who was found to have massive inguinoscrotal herniation. Preoperative computerized tomography showed bilateral hydronephrosis. Herniorrhaphy was successfully done through an inguinal incision. The left kidney regained normal function with a decrease in creatinine levels. This case is atypical because of the presence of massive inguinoscrotal herniation with bilateral hydronephrosis. Computerized tomography has proved to be a valuable tool in diagnosing such cases. Surgical repair and reduction of hernia can be accomplished through an inguinal incision. PMID- 8230534 TI - Severe cyclophosphamide-induced hemorrhagic cystitis successfully treated by total cystectomy with ileal neobladder substitution: a case report. AB - A 45-year-old woman with intractable cyclophosphamide-induced hemorrhagic cystitis was successfully treated with total cystectomy and ileal neobladder substitution. To our knowledge this is the first reported reconstruction of the lower urinary tract in a patient with acute hemorrhagic cystitis using a neobladder. Neobladder substitution is contraindicated if the urethra or bladder neck is involved in the disease, although neither was involved in our patient. Whether these lesions are generally left intact has not been discussed previously. If a neobladder can be used, life threatening hemorrhagic cystitis should be treated with total cystectomy accompanied by immediate neobladder substitution. PMID- 8230535 TI - Carcinoma in female urethral diverticulum: case reports and review of management. AB - We report on 4 women ages 31 to 65 years with adenocarcinoma of urethral diverticulum. One patient presented with metastatic disease and died after refusing post-radiation surgery. The other 3 women were without evidence of disease 9 to 44 months after diverticulectomy with radiation (1) or radical surgery (2). Review of our 4 cases and 64 cases documented in the literature showed that irritative voiding symptoms and hematuria were the most common findings. A palpable urethral mass was described in only 56% of reported cases but was observed in all of our patients, with the smallest mass measuring 2 x 2 cm. Urethroscopy with biopsy is the most important diagnostic test. Aggressive treatment is appropriate in most patients, since local recurrence has been documented in 44% of those treated by diverticulectomy and/or radiation. PMID- 8230536 TI - Isolated amyloidosis of the penile urethra and corpus spongiosum: a case report. AB - Localized amyloidosis of the penile urethra and corpus spongiosum is rare. The pathogenesis is obscure and optimal management of such patients has not been defined. We report a case that was treated with transurethral removal of amyloid tissue, which was obstructing the urethral outlet. Previously, the patient had been treated with dilation with temporary relief of the symptoms. He was disease free 1 1/2 years postoperatively. PMID- 8230537 TI - Scrotal arteriovenous malformation: case report and literature review. AB - We report a rare case of scrotal arteriovenous malformation in an adolescent. Arteriovenous malformation of the scrotum is distinguished radiographically and histologically by numerous arterioles and thick walled veins with no intervening capillary bed. The cardinal sign is a bruit. Complete excision is recommended as definitive treatment. Preoperative angio-embolization may be helpful but it should not be relied upon for clinical resolution. PMID- 8230538 TI - Giant multilocular prostatic cystadenoma: a rare clinical entity and review of the literature. AB - Giant multilocular prostatic cystadenoma is a pathologically benign entity. To our knowledge only 3 cases have been reported in the literature, of which 2 were treated by means of pelvic exenteration. These lesions can occur in men of various ages, arise from the prostate and grow to massive proportions. Histologically the tumor is comprised of benign cysts lined with cuboidal and columnar epithelium. The lesions do not invade contiguous structures but they can be adherent to viscera in their proximity. Giant multilocular prostatic cystadenoma as demonstrated by our case is a benign entity that can be definitively treated by carefully planned complete surgical excision. PMID- 8230539 TI - Intravesical instillation of terodiline--an in vivo study of drug absorption in rabbit and man. AB - The absorption of intravesically administered terodiline was investigated in 8 healthy volunteers (10(-4)) and in 33 rabbits (10(-4)) M., 4 X 10(-4) M. and 1.6 X 10(-3) M.). In the humans, the amount of drug retained and the serum concentration were registered. In the rabbits, the influence of drug concentration, pH (5 and 9.2) and volume (5 and 20 ml.) were evaluated. Liquid scintigraphy of the bladder wall and of serum and tissue samples from striated muscle, kidney and liver was performed. Further, autoradiography of the bladder wall and calculation of drug retention were done. In the humans, 20 +/- 5% of the administered terodiline dose was retained in the body. Clinically significant serum concentrations were not detected. In the rabbits, 21 +/- 4% of the drug was retained with the pH = 5 terodiline solution as compared with 64 +/- 3% with the pH = 9.2 solution. With increasing terodiline concentrations, drug retention increased, as did the tissue and serum activities. At a given concentration, the total amount of drug retained increased with a larger volume. The drug gradient between the bladder muscle layer and serum was of a magnitude of about 100, independent of the drug concentration within the muscle layer. It is concluded that the pH and concentration of the solution are factors of major importance for the amount of terodiline absorbed from the bladder. Compared with the one obtainable by systemic drug administration, the large gradient between the bladder muscle and the serum is conspicuous, and may be of clinical interest. PMID- 8230540 TI - Dissecting the immunobiological effects of Bacillus Calmette-Guerin (BCG) in vitro: evidence of a distinct BCG-activated killer (BAK) cell phenomenon. AB - Several immunobiological effects of intravesical bacillus Calmette-Guerin (BCG) during immunotherapy of superficial bladder cancer have been suggested as possible mediators of the mode of action. In an attempt to elucidate which of these effects is relevant to tumoricidal activity, an in vitro cytotoxicity assay was employed in which the direct effects of BCG and of cytokines against four transitional carcinoma cell lines were studied. Furthermore, peripheral blood mononuclear cells (PBMNC) were analyzed for their cytotoxic potential against these target cells. We found that none of the cytokines interleukin-1, interleukin-2, interferon-gamma, tumor necrosis factor alpha, lymphotoxin, or BCG alone were cytotoxic against the bladder carcinoma cell lines. However, a pronounced cytotoxicity against these targets resistant to natural killer cells could be induced in PBMNC by coincubation with viable BCG. We termed this the BCG activated killer (BAK) cell phenomenon. In contrast to lymphokine-activated (LAK) cells, these BAK cells needed prolonged activation for 7 days and did not enhance the cytotoxicity against K562 target cells sensitive to natural killer cells. Nonviable, heat-inactivated BCG was significantly less effective, and sonificated fractions of BCG were not effective in stimulating PBMNC towards BAK cell activity. In vitro dissection of effects observed during BCG intravesical therapy may give more insight into the mode of action of BCG and may help to separate primary tumoricidal effector mechanisms from secondary concomitant phenomena. Further characterization of the BAK cell may result in an improvement of intravesical BCG immunotherapy. PMID- 8230541 TI - Endogenous factors modulating urinary bladder compliance in mice. AB - The application of repeated cystometric measurements in longitudinal long-term studies both in patients and experimental animals requires detailed knowledge of factors causing intraindividual variability. In the present study, the effect of age, body weight, estrus cycle phase, rectal temperature and premeasurement bladder volume on the results of cystometry in mice have been assessed. No systematic changes in bladder compliance parameters were observed with age (75 to 344 days) or body weight (18 to 38 gm.). Also, the animal's position in the estrus cycle had no effect on bladder capacity. The abdominal temperature, adjusted to 32 to 37.3C under anesthesia, did not affect the outcome of cystometry either. When the bladder was emptied at 1.5 to 3.5 hours before cystometry, a significant drop in bladder capacity to about 60% of individual control values was observed. This initial emptying clearly decreased the intraindividual variability in absolute terms (standard deviation), but not the relative variability (coefficient of variance). Thus only a shift in absolute values, but no improvement in reproducibility of cystometric results, could be achieved. Bladder filling to 10 or 25 mm. Hg 40 minutes to 2 hours before cystometry increased the bladder capacity to 120 or 145%, respectively, when anesthesia was maintained throughout the experiment. When intervals longer than 2 hours were studied, animals were allowed to regain consciousness between initial bladder filling and cystometry. Under these conditions, the capacity data returned to normal or subnormal values. PMID- 8230542 TI - Effects of nerve stimulation on blood flow in the urinary bladder, urethra and pelvic floor in the dog. AB - Pelvic organs have triple innervation from the pelvic, sympathetic and pudendal nerves. Peripheral effects can be studied by neurostimulation of the nerves, whereas the topography of the spinal cord neurons can be determined by horseradish peroxidase tracing. We have evaluated the neurophysiologic effects of the nerves at their target organs by intraoperative electric stimulation and compared the effects to the anatomical innervation areas. These zonal areas were determined by blood flow alterations, measured with nine different microspheres. Pelvic nerve stimulation showed bladder contraction with no significant blood flow changes during stimulation. Sympathetic nerve stimulation caused moderate detrusor contraction and pressure increase in the bladder neck and intraprostatic area. Regional blood flow showed a four-fold increase in the bladder neck area during neurostimulation. Pudendal nerve stimulation revealed an intraurethral pressure increase with a 3.5-fold increase of blood flow in the sphincteric area and in the pelvic floor musculature. PMID- 8230543 TI - In vitro sensitivity testing of human bladder cancers and cell lines to TP-40, a hybrid protein with selective targeting and cytotoxicity. AB - TP-40 is a hybrid fusion protein produced by recombinant technology and consists of a molecule of transforming growth factor-alpha (TGF-alpha) fused to the Pseudomonas exotoxin PE-40. A panel of human and murine bladder cancer cell lines was found to be universally sensitive in vitro to TP-40 in a clonogenic assay. All lines expressed receptor for epidermal growth factor (EGF), though none demonstrated gene amplification for the EGF receptor. The sensitivity to TP-40 may be blocked by preexposure to EGF. Six human bladder tumors taken directly from patients were all sensitive in vitro to TP-40; these included well differentiated tumors. TP-40 may prove to be effective as an intravesical agent in bladder cancer via selective targeting to cells that express EGF receptors, as do the majority of human bladder cancers. PMID- 8230544 TI - Laser-assisted enterocystoplasty in rats. AB - The employment of laser welds to anastomose bowel to bladder in urological surgery has never been described. Using rats, we compared laser-assisted enterocystoplasty (LAE) with conventional sutured enterocystoplasty (CSE). A "patch" ileocystoplasty was performed using distal ileum. Operative time, intraluminal bursting pressure and histologic changes were evaluated. Operative time was 38% less in the LAE (p < 0.001). No statistically significant differences in bursting pressure between LAE and CSE groups was demonstrated (p = 0.654) in rats sacrificed 2 weeks postoperatively. Histologic findings in the laser-assisted enterocystoplasties included early loose granulation tissue and later organization of the granulation tissue with significant collagen deposition. In our preliminary study, laser welding of bowel within the urinary stream was safe, efficacious and rapid and resulted in a mechanically comparable surgical outcome. PMID- 8230545 TI - Electrophysiological study of relations between the dorsal nerve of the penis and the lumbar sympathetic chain in the rat. AB - Afferent sensory inputs from the penis are carried by the dorsal nerve of the penis (DNP) to the spinal cord. Sympathetic outflow involved in the control of the urogenital tract is partly conveyed by the lumbosacral sympathetic chain. Our aim was to search for a sympathetic component in the DNP and relations between DNP afferents and sympathetic fibers conveyed by the distal sympathetic chain in anesthetized adult male rats. Stimulation of the lumbar sympathetic chain at the L4-L5 level (LSC4-5) elicited an evoked discharge on the DNP. This discharge was abolished by cutting the sympathetic chain distal to the stimulation site. Ganglionic blockade with hexamethonium and various neural sections revealed the presence of sympathetic postganglionic fibers in the DNP, originating in the sympathetic chain. Stimulation of the DNP evoked a reflex discharge in the LSC. This reflex was spinally mediated since it was abolished by acute spinal cord transection at the L5 level. Acute spinalization at the T8 level significantly reduced the latency of the evoked response. We hypothesize that both spinal and supraspinal control exist over relations of the DNP afferents with sympathetic outflow to the pelvis. Increase in sympathetic tone elicited by activation of penile sensory fibers could play a role in regulation of sexual function. PMID- 8230546 TI - Bladder damage in mice after single and repeated intravesical instillations of mitomycin C or doxorubicin. AB - Functional and histological damage after intravesical administration of mitomycin C or doxorubicin were studied in mouse bladders. Dosing schedules were based on those commonly used in the clinic, and bladder function was assessed from changes in urination frequency. Repeated weekly instillations of 1 mg./ml. mitomycin C induced a transient three-fold increase in Frequency Index (FI) with recovery to control levels within 3 weeks. Weekly instillations of 6 mg./ml. doxorubicin also resulted in a three-fold increase in FI, but lower doses produced only minimal changes. There was, again, full recovery within 3 weeks. Our experiments indicate that repeated intravesical instillations of mitomycin C or doxorubicin are well tolerated, with rapid recovery from functional damage within a few weeks of cessation of therapy. There was significantly less damage after repeated instillations of clinically relevant doses of doxorubicin than mitomycin C. PMID- 8230547 TI - Reduction of drug accumulation in cisplatin-resistant variants of human prostatic cancer PC-3 cell line. AB - We have isolated cis-diamminedichloroplatinum (II) (CDDP)-resistant variants, P/CDP4 and P/CDP5, from human prostatic cancer PC-3 cells after a stepwise exposure to CDDP. P/CDP4 and P/CDP5 showed 11-fold and 23-fold higher resistance to CDDP than did PC-3. P/CDP5 was cross-resistant to carboplatin, mitomycin C, etoposide, m-AMSA, bleomycin and UV irradiation. Alkaline elution of DNA showed an increased amount of DNA interstrand cross-links in PC-3 but not in P/CDP5 when PC-3 and P/CDP5 were cultured with CDDP. Flameless atomic absorption spectrophotometry revealed that intracellular accumulation of CDDP in P/CDP4 and P/CDP5 was decreased to 18 to 34% and 9 to 18% of that of PC-3, respectively, when PC-3 and its CDDP-resistant counterparts were incubated with 5 and 10 micrograms./ml. of CDDP for 24 hours. These data suggest that decreased drug accumulation is involved in the development of CDDP-resistance in the PC-3 cell line. PMID- 8230548 TI - Treatment of a human renal cell carcinoma in nude mice with recombinant human tumor necrosis factor alpha and etoposide. AB - The effect of treating a human renal cell adenocarcinoma xenografted into Balb/c nu/nu (nude) mice with recombinant human tumor necrosis factor alpha (TNF alpha) and the cytostatic agent etoposide (ETP) as monotherapy or combination has been studied. Antitumor effects were evaluated by determining growth of the tumor implants by external caliper measurements and tumor cell proliferation by determining the labelling index (LI) after pulse labelling with 3H-thymidine. The toxicity of the treatment with TNF alpha and/or ETP was also studied by measuring the animal weight. Monotherapy with TNF alpha had no effect on tumor growth or proliferation. Treatment with ETP as a single agent, TNF alpha plus ETP applied concurrently and TNF alpha plus ETP two days later led to a slight inhibition of tumor growth and also to a slight decrease of the LI. In contrast to a monotherapy with TNF alpha, all therapeutic modalities containing ETP showed an increased toxic effect on the animals represented by a distinct weight loss. This suggests that the minute efficacy of the treatment observed could well be due solely to its toxicity. In contrast to two other studies, no additive or synergistic effect of the antineoplastic activity of TNF alpha and/or ETP was found. The intertumoral variation of human renal cell carcinomas could be one reason for the different results with this therapeutic regimen. PMID- 8230549 TI - Scanning electron microscopic study of the effect of citrate and pyrophosphate on calcium oxalate crystal morphology. AB - The effects of citrate and pyrophosphate on the morphology of calcium oxalate monohydrate crystals formed in a supersaturated calcium oxalate solution were examined by scanning electron microscopy. Within the normal range of urinary concentrations, citrate and pyrophosphate affected the morphology of the crystals. The length/width ratio of the crystals decreased at higher concentrations of citrate and pyrophosphate. Citrate seems to have a dual mechanism of inhibition of crystal growth (binding to the crystal surfaces) and of complexation (with calcium in the solution). Pyrophosphate is believed to bind preferentially to calcium in the solid phase rather than in the solution phase, as it affected crystal morphology at very low concentrations and was detected on the crystals by X-ray microanalysis. Pyrophosphate seems to bind to the monohydrate but not to the dihydrate. PMID- 8230550 TI - Long-term effects of ureteric stent after ureteric dilation. AB - Balloon dilation of the right ureterovesical junction (UVJ) and distal ureter to three times its normal caliber was performed in 12 pigs. A right double-J (D-J) stent was inserted after dilation in 6 pigs. Bilateral upper tract dynamics with different perfusion rates (0.5, 2 and 4 ml. per minute) were recorded before dilation, immediately after dilation, and then 4 and 7 weeks after dilation. Immediate and late antegrade nephrostograms as well as suprapubic cystograms were taken. Grade 3 reflux occurred in 100% of animals at 7 weeks on the dilated, stented ureter and no reflux on the dilated, nonstented ureter. At 7 weeks on the dilated, stented side, significant growth (> 100,000, colonies) of Pseudomonas species was noted in all animals. Creatinine clearance was significantly reduced on the dilated, stented side when compared to the dilated, nonstented side at 7 weeks. Histologic examination of the dilated, stented and dilated, nonstented ureters at 4 weeks revealed a segmental muscular defect with muscular regeneration starting from the edge of the defect, particularly in the innermost region. At 7 weeks, there was a more advanced, but similar, pattern of muscular regeneration in both groups. However, at 7 weeks, metaplastic changes of the ureter and chronic pyelonephritis were evident on the dilated, stented ureter. Electron microscopy showed that myofibroblasts played a major role in the healing process with new muscle formation. At 4 weeks, no significant morphologic difference was found between the dilated, stented and dilated, nonstented ureters. At 7 weeks, however, it appeared that the ureteric stent resulted in damage and deterioration of renal function without affecting muscular regeneration of the ureter. We conclude that the changes observed could be entirely due to the infection associated with the stent rather the stent itself. PMID- 8230551 TI - Evaluation of direct bladder stimulation with stainless steel woven eye electrodes. AB - Encouraged by recent clinical reports of micturition induced in patients by direct bladder stimulation, we conducted a study of optimum methods of direct bladder stimulation. During surgery six male cats received eight large surface area woven eye electrodes sutured to the bladder wall serosa, four on the bladder dome and four adjacent to the trigone area. Two additional small surface-area single knot electrodes were sutured in the trigone area. Suprapubic and intraperitoneal tubes were placed for pressure recording and bladder filling. Leg and pelvic floor EMG electrodes were also used for tethered recordings. One to eight weeks after surgery, optimum stimulation methods were evaluated as the animal freely moved about a urodynamic recording cage. Electrodes in the trigone region were more effective than electrodes on the dome and induced bladder contractions and voiding similar to spontaneously induced voiding with bladder filing. Large surface area, woven eye electrodes, composed of multistranded 316LVM stainless steel wire, were more effective than smaller surface area single knot electrodes. High stimulating frequencies (40 Hz) were better than lower frequencies (10 to 20 Hz), and a 1 millisecond pulse duration was optimal. Pulsing with stimulating currents from 10 to 25 mA induced effective bladder contractions with voiding when applied for 3 seconds. However, lower currents using longer stimulation periods were also effective. Bipolar electrodes with both electrodes on the bladder wall were superior to monopolar arrangements with the positive ground electrode along the animal's back. We concluded that in the able-bodied cat model, bladder contractile activity for micturition can be induced with direct bladder stimulation and with little discomfort. An effective stimulation protocol consists of capacitor-coupled monophasic pulses with large surface area bipolar electrodes in the trigone region. Stimulating parameters of 40 Hz, 1 msec., 10 to 25 mA applied for 3 seconds were optimal. In addition, based on corrosion resistance observations, the electrodes are quite suitable for long-term studies. PMID- 8230552 TI - Restoration of spermatogenesis and subsequent fertility by direct intratesticular hormonal therapy. AB - Testosterone-laden microspheres (TLM) were used in an attempt to restore spermatogenesis and sustain fertility in rats with previously suppressed gonadal function. Adult rats received 62.5 micrograms. gonadotropin-releasing hormone antagonist (GnRH-antagonist) per day for 15 days prior to being divided into three groups: 1) animals to be examined immediately, 2) animals to continue receiving 62.5 micrograms. GnRH-antagonist per day alone, and 3) animals to continue receiving GnRH-antagonist + a single intratesticular injection of 20 mg. TLM per testis. Unoperated control and sham-operated animals were also studied. Testicular interstitial fluid (TIF) testosterone concentrations, testicular sperm production and fertility were assessed 105 days after initiation of the experiment. Testicular interstitial fluid testosterone concentrations and sperm production were significantly reduced from control values after the initial 15 day GnRH-antagonist treatment (p < .05). After 105 days of GnRH-antagonist treatment, these values were further reduced (p < .05), but the TIF testosterone and sperm production values of GnRH-antagonist treated animals receiving TLM supplementation were not different from those of controls (p < .05). Fertility trials were also performed. Animals which had received GnRH-antagonist alone for 90 days were completely infertile, but animals receiving GnRH-antagonist + TLM for 90 days had fertility values not different from those of control animals (p < .05). Thus, spermatogenesis reduced because of testicular insufficiency was restored to normal, as was fertility, by intratesticular administration of TLM. This technique has possible clinical applications in selected groups of infertile men. PMID- 8230553 TI - Characterization and localization of prostatic alpha 1 adrenoceptors using radioligand receptor binding on slide-mounted tissue section. AB - Alpha 1 adrenoceptor binding sites have been characterized in prostatic tissue homogenates using radioligand receptor binding studies. The objective of the present study was to characterize and localize prostatic alpha 1 adrenoceptor binding sites using slide-mounted tissue sections and the ligand 3H-prazosin. The present study demonstrated that preincubation is not required; the optimal incubation interval is 40 minutes; and a 1-minute wash (once or twice) maximizes the proportion of specific 3H-prazosin binding. Saturation studies were performed at 8 different concentrations of 3H-prazosin ranging between 0.0625 nM. to 8.0 nM. The binding of 3H-prazosin was consistently saturable and of high affinity. The mean Kd and Bmax determined from 6 saturation studies was 4.16 x 10(-10) M. and 1.30 fmol./mg. wet weight, respectively. The pharmacology of these 3H prazosin binding sites was characterized using competitive displacement experiments. The mean IC50 corrected for prazosin, phentolamine and yohimbine was 7.8 x 10(-10) M., 6.0 x 10(-9) M. and 2.1 x 10(-6) M. The rank order of the IC50 corrected values indicates that alpha 1 binding sites were measured under the assay conditions. In the present study, the mean values for Kd, Bmax and IC50 corrected are similar to values previously reported using prostatic tissue homogenates. Prostatic tissue sections were apposed to x-ray film after being incubated with 3 nM. 3H-prazosin (total prazosin binding) and 3 nM. 3H-prazosin + 8 microM. prazosin (nonspecific prazosin binding). The autoradiograms were analyzed using a computerized analyzing system. The specific radioactive densities of 3H-prazosin in the stroma and glandular epithelium were 1099 +/- 48 pCi/mg. and 163 +/- 42 pCi/mg. The present study validates the technique of assaying alpha 1 adrenoceptor binding sites on slide-mounted prostatic tissue sections and provides further evidence that alpha 1 adrenoceptor binding sites are localized primarily to the stromal elements of the prostate. PMID- 8230554 TI - Evidence for purinergic neurotransmission in human urinary bladder affected by interstitial cystitis. AB - Detrusor specimens were obtained from 5 patients affected by interstitial cystitis (IC) and 5 patients with bladder carcinoma (controls). Muscle strips were prepared for in vitro pharmacological studies. In all detrusor strips taken from IC patients, an important portion of the electrically-induced contraction was atropine-resistant. In contrast, atropine-resistance was never observed in control detrusors. H1 and H2 antagonists did not affect noncholinergic contractile response which, conversely, was abolished following desensitization to alpha, beta methylene ATP (APCPP). Detrusor muscle from patients affected by IC exhibited an increase in sensitivity to APCPP and a decrease in sensitivity to acetylcholine with respect to control detrusor. Taken together these results are consistent with the presence of a purinergic neurotransmission in parasympathetic nerve terminals of the urinary bladder affected by IC, probably as a consequence of alterations in the innervation and/or electrical coupling between smooth muscle cells. The sensitivity of IC detrusor muscle to histamine was much lower than that of control detrusor, suggesting a desensitization of histamine receptors present in the bladder wall of IC patients. PMID- 8230555 TI - Aberrant expression of gangliosides in human renal cell carcinomas. AB - Aberrant and elevated ganglioside expression has been observed in neoplasms, and has been shown to be an important marker of tumor progression. We therefore studied the gangliosides of renal cell carcinoma (RCC) by analyzing gangliosides from 18 RCC biopsies, 10 RCC lines and 5 normal kidney biopsies. A comparison of tumor with normal tissue revealed a significant difference in individual ganglioside expression in which the former consistently expressed eight major gangliosides, GM3, GM2, GM1, GD3, GD1A, GD2, GD1B and GT1B, according to the nomenclature of Svennerholm. There was a notable significant mean increase in the expression of GM2, GM1 and GD1A and a significant decrease in the expression of GD3 in tumor tissue compared with normal kidney tissue. Compared with tumor biopsy tissue, RCC cell lines showed a significant decrease in the expression of GM3, but a significant increase in GM2, GM1 and GD2. There was a marked increase in a pathway gangliosides (GM2, GM1, and GD1a) in RCC biopsies and cell lines compared with normal kidney. These studies indicating that RCC have markedly aberrant ganglioside expression similar to neural origin tumors may relate to the activation of a ganglioside pathway enzymes. Gangliosides expressed on RCC tumors may be important markers of tumor progression and target antigens for immunotherapy. PMID- 8230556 TI - The AVF, FDA, AMA, AAMC, ACCME, and CME. PMID- 8230557 TI - Magnetic resonance venography for the detection of deep venous thrombosis: comparison with contrast venography and duplex Doppler ultrasonography. AB - PURPOSE: Contrast venography is the gold standard for diagnosis in deep venous thrombosis (DVT); however, this technique is invasive and requires the use of potentially hazardous contrast agents. Although duplex Doppler ultrasonography is accurate in the evaluation of lower extremity DVT, it is less accurate in the assessment of the pelvic and intraabdominal veins. Magnetic resonance venography (MRV) has recently been developed, and our purpose was to determine whether MRV could accurately demonstrated DVT when compared with duplex scanning and contrast venography. METHODS: Eighty-five patients underwent contrast venography and MRV from the inferior vena cava to the popliteal veins to rule out DVT. Thirty-three of these patients also underwent duplex scanning. Blinded readings of these studies were compared for the presence or absence and extent of venous thrombosis. RESULTS: DVT was documented by contrast venography in 27 (27%) venous systems. Results of MRV and contrast venography were identical in 98 (97%) of 101 venous systems, whereas results of duplex scanning and contrast venography were identical in 40 (98%) of 41 venous systems. All DVTs identified by contrast venography were detected by MRV and duplex scanning. The discrepancies were due to false-positive MRV (3) and duplex scanning (1) results. When compared with contrast venography, MRV had a sensitivity of 100%, specificity of 96%, positive predictive value of 90%, and negative predictive value of 100%. For duplex scanning the sensitivity was 100%, specificity was 96%, positive predictive value was 94%, and negative predictive value was 100%. CONCLUSIONS: It is concluded that MRV is an accurate noninvasive venographic technique for the detection of DVT. PMID- 8230558 TI - Refinements in the ultrasonic detection of popliteal vein reflux. AB - Color-flow and duplex ultrasonography were used to determine the optimal method for documenting venous valvular reflux. Popliteal veins were examined in 10 normal limbs and 11 limbs with clinical evidence of chronic venous insufficiency (CVI). Peak reflux velocity (spectral) and duration of reflux (spectral and color) were measured with the patient in supine and standing positions, with manual and pneumatic compression applied sequentially to thigh and calf. Manual and pneumatic compression produced equivalent reflux velocity and duration. In normal limbs peak reflux velocity was always less than 22 cm/sec, with a mean reverse flow duration of 0.3 sec +/- 0.03 (SEM). In limbs with CVI, reflux velocity varied widely among protocols. Reflux duration and velocity were greater in the supine position than in the standing position for both normal limbs and limbs with CVI (p < 0.04). Duration was significantly increased for thigh versus calf compression in normal limbs (p < 0.001) but decreased in limbs with CVI (p < 0.003). Methods that used thigh compression or supine position were less capable of discriminating normal limbs from limbs with CVI. Standing calf compression provided the greatest rates of sensitivity (91%), specificity (100%), and accuracy (95%). Compared with spectral Doppler scanning, color-flow ultrasonography produced a consistently shorter reflux duration (p < 0.001). In limbs with CVI with a mean spectral duration of 2.5 sec +/- 0.2 (SEM), mean color Doppler duration was 0.7 sec shorter. Our results demonstrate that popliteal vein incompetence is identified optimally by reflux duration after standing calf compression; adequate manual compression is sufficient to identify reflux; color flow Doppler ultrasonography may underestimate reflux duration. PMID- 8230559 TI - Influence of age on venous physiologic parameters. AB - PURPOSE: The incidence of deep venous thrombosis increases significantly with age. Attempts to explain this association have failed to identify factors that could be contributory. We hypothesized that age-related changes in venous physiologic parameters might contribute to an increased risk of deep venous thrombosis. METHODS: Air plethysmography was used to measure a number of physiologic parameters in two sets of patients. Group A consisted of 17 subjects (33 limbs), age range 23 to 40 years. Group B consisted of 11 subjects (22 limbs), age range 60 to 83 years. No subject in either group had peripheral vascular disease or a history of deep venous thrombosis. Parameters measured were venous volume, venous filling index, ejection volume, ejection fraction, residual volume fraction, and outflow fraction. RESULTS: Venous volume and ejection fraction were significantly reduced in Group B compared with Group A. Residual volume fraction was significantly increased in Group B compared with Group A. There were no statistically significant differences in outflow fraction, venous filling index, or ejection fraction between the two groups. CONCLUSIONS: These results suggest that the efficiency of the calf muscle pump diminishes with increasing age, possibly contributing to the increased incidence of deep venous thrombosis in the elderly. PMID- 8230560 TI - Distribution and quantification of venous reflux in lower extremity chronic venous stasis disease with duplex scanning. AB - PURPOSE: The purpose of this study was to use color-flow duplex scanning to identify the anatomic distribution of venous reflux and to quantify venous reflux times in patients with various stages of chronic venous insufficiency (CVI). METHODS: Color-flow-assisted duplex scanning was used to identify the anatomic distribution of venous reflux and to quantify reflux times in the deep and superficial venous systems of patients with symptomatic (CVI). Two hundred two patients with class I to III CVI were examined. RESULTS: Only 11% (22 patients) had a documented history of phlebothrombosis. Of the 403 limbs evaluated, 192 had venous ulcers whereas 211 were classified as having class I or II CVI. Nonocclusive venous obstruction was found in only 16 limbs (4%). Venous ulceration was significantly associated with reflux in multiple venous segments as opposed to reflux in isolated venous segments (p < 0.001). Total limb reflux time (Rt) was determined by summing the reflux times of all the venous segments in a limb. The mean Rt of patients with venous ulcerations was significantly longer than the mean Rt of limbs with class I and II CVI (p < 0.01). A total limb reflux time of greater than 9.66 seconds was predictive of ulceration. Total limb deep segment reflux time and total limb superficial segment reflux time were also determined by summing the reflux times of the appropriate segments in the limb. The mean deep segment reflux time was prolonged in limbs with venous ulcers when compared with limbs with class I and II CVI disease. The mean superficial segment reflux time of limbs with class I and II CVI and limbs with venous ulcers could not be used to distinguish between the two groups. In assessing the contribution of segments of the deep system to ulceration, reflux times of different segments were compared with wound duration and area. Reflux in the common femoral vein was significantly associated with wound area and duration (p < 0.05) whereas reflux time in the distal posterior tibial vein was associated with wound duration (p < 0.05). PMID- 8230561 TI - Popliteal vein entrapment presenting as deep venous thrombosis and chronic venous insufficiency. AB - PURPOSE: This report describes popliteal vein entrapment in three patients and demonstrates that it may present with manifestations of typical venous disease. METHODS: This report was compiled from a review of inpatient and outpatient records. RESULTS: In the first case, a 28-year-old woman was seen with left leg popliteal and calf deep vein thrombosis without obvious cause. She described long standing calf discomfort, and passive dorsiflexion of the left foot caused disappearance of arterial pulsations at the pedal level. She was given the anticoagulants heparin and sodium warfarin (Coumadin) followed by surgical exploration. The popliteal vein and artery were entrapped by a fibrous extension of the medial head of the gastrocnemius muscle attaching to the lateral femoral condyle. After band lysis, the patient has been symptom free for 6 years. The second patient, a 37-year-old man, was seen with bilateral chronic venous insufficiency (CVI). Passive dorsiflexion and active plantar flexion of the feet did not diminish the pedal pulses; impedance plethysmography suggested mild outflow obstruction. Ascending venography demonstrated entrapment at the midportion of duplicated popliteal veins with no postthrombotic changes. He was treated with compression stockings and has done well during an 18-month follow up. The third patient, a 17-year-old male, was seen with severe symptoms of right leg CVI and venous obstruction since 3 years of age. Air plethysmography revealed ambulatory venous hypertension, whereas venography demonstrated reflux down to the knee with an extrinsic narrowing at the midpopliteal vein. During operation, an abnormal origin of the lesser saphenous vein (LSV) from the popliteal vein was found; the LSV took a medial route, compressed the tibial nerve, and caused severe distortion and narrowing of the popliteal vein. Division of the LSV resulted in release of popliteal venous compression and immediate relief of symptoms. CONCLUSIONS: The three cases presented demonstrate that popliteal venous entrapment may begin with symptoms of deep vein thrombosis and CVI. Popliteal venous entrapment must be considered in the differential diagnosis of venous disease in younger patients in whom common predisposing factors are absent. PMID- 8230562 TI - Light reflection rheography: a simple noninvasive screening test for deep vein thrombosis. AB - PURPOSE: The clinical diagnosis of deep vein thrombosis (DVT) is unreliable. Contrast phlebography (CP) continues to be the gold standard, but it is invasive. Although duplex ultrasonography is an accurate, noninvasive alternative, it is expensive, technically demanding, and time-consuming. We postulated that light reflection rheography (LRR), a noninvasive method of assessing the quantity and rate of venous emptying, might be a reliable and inexpensive bedside approach to screening patients with clinically suspected DVT. METHODS: With LRR, infrared light is beamed onto the skin, and the amount of backscattered rays are detected, which indirectly measures the amount of blood present in a volume of the epidermis beneath the LRR probe. Applied to the calf muscle pump, LRR can provide a noninvasive method of assessing blood volume changes in the sample area of skin, in response to venous hemodynamic changes in the lower limb. RESULTS: Sixty nine limbs in 61 patients undergoing CP for clinically suspected DVT over a period of 12 months also underwent LRR, either just before or within 24 hours of undergoing phlebography. The criteria for diagnosing DVT on CP were presence of filling defect or nonfilling of a venous segment. The result of LRR was considered positive for DVT if the rate of venous emptying was 0.35 or less. With these criteria a sensitivity of 96.4% and specificity of 82.9 were obtained. This resulted in a positive predictive value of 79% and a negative predictive value of 97.1%. CONCLUSIONS: LRR is a simple, inexpensive, and noninvasive bedside test that takes 10 minutes to perform. It is highly sensitive with a high negative predictive value, detecting most cases of DVT, reliably ruling out DVT, and eliminating the need for more time-consuming and costly studies. Therefore it seems to be an appropriate screening test in patients with clinically suspected DVT. PMID- 8230563 TI - Noninvasive evaluation of the lymphatic system with lymphoscintigraphy: a prospective, semiquantitative analysis in 386 extremities. AB - PURPOSE: Lymphoscintigraphy has emerged as the diagnostic test of choice in patients with suspected lymphedema. To assess the lymphatic circulation of 386 extremities in 188 patients, we prospectively recorded a semiquantitative index of lymphatic transport in addition to visual evaluation of lymphoscintigraphy image patterns. METHODS: Sixty-one male and 127 female patients were studied (mean age 48 years, range 13 to 87 years). Twenty had upper extremity swelling, and 168 had lower extremity swelling. The disease was bilateral in 60 patients. Lymphoscintigraphy was performed by injecting a mean of 503 microCi of technetium 99m-antimony trisulfide colloid subcutaneously into the second interdigital space of the extremity. Time for transport to regional lymph nodes, appearance of lymph vessels and nodes and distribution pattern were scored. These scores were compiled into a modified Kleinhans transport index (TI). To assess the venous circulation, 155 patients underwent evaluation of the venous system by impedance plethysmography, ultrasonography, or contrast venography. RESULTS: The mean TI (+/- SEM) in 79 asymptomatic extremities was 2.6 +/- 0.5, with 66 (83.5%) demonstrating normal lymphoscintigraphy pattern (TI < 5). Patients with clinical diagnosis of lymphedema (n = 124) had a mean TI of 23.8 +/- 1.5; 81.5% of these were greater than 5. Fifty-six patients (30%) had primary and 68 (36%) had secondary lymphedema. (TI of 26 +/- 3.5 and 22.1 +/- 1.9, respectively, p = NS). Patients without any lymphatic transport (TI of 45) were more likely to have cellulitis in their history (p < 0.05). Contrast lymphangiography in six patients correlated with lymphoscintigraphy. Sixty-four patients (34%) had swelling without lymphedema (venous edema, cardiac edema, lipedema, etc.; TI of 1.9 +/- 0.4, p < 0.001). Of the 41 patients with abnormal venous studies, 18 (44%) had an elevated TI. CONCLUSIONS: Semiquantitative evaluation of the lymphatic transport with lymphoscintigraphy reliably depicts abnormalities in the lymphatic circulation. Lymphoscintigraphy excluded lymphedema as a cause of leg swelling in one third of our patients. PMID- 8230564 TI - Perimalleolar subcutaneous tissue pressure effects of elastic compression stockings. AB - PURPOSE: We hypothesized that the clinical benefit of elastic compression stockings (ECS) is at least in part due to an increase in subcutaneous pressure that may promote resorption of extracellular fluids, providing more efficient diffusion of oxygen and nutrients from the microcirculation to the skin and subcutaneous tissues. METHODS: To test this hypothesis we designed and standardized a device for measuring subcutaneous pressure in patients. We then measured the supine perimalleolar subcutaneous pressure from a single limb in four groups: group 1 consisted of control subjects (n = 8); group 2 consisted of patients with varicose veins and superficial venous insufficiency without lipodermatosclerosis or edema (n = 5); group 3 consisted of patients with deep venous insufficiency and lipodermatosclerosis but without edema (n = 8); and group 4 consisted of patients with deep venous insufficiency, lipodermatosclerosis, and clinically evident edema (n = 8). Measurements were made at baseline and after application of 20 to 30 mm Hg and 30 to 40 mm Hg ECS. RESULTS: There was no significant difference in the baseline subcutaneous pressure between the three groups without clinical edema (p > 0.05). Baseline perimalleolar pressure was elevated, however, in group 4 patients compared with groups 1, 2, and 3 (p < 0.05). All three groups with chronic venous insufficiency (CVI) (groups 2, 3, 4) demonstrated increases in subcutaneous pressure with application of ECS, which was statistically significant in groups 3 and 4. There was no difference between the increase in perimalleolar subcutaneous pressure induced by 20 to 30 mm Hg or 30 to 40 mm Hg ECS in groups 3 and 4. CONCLUSIONS: Patients with CVI and edema have significant elevations in supine resting perimalleolar subcutaneous pressure compared with control subjects and patients with CVI without edema. Twenty to 30 mm Hg and 30 to 40 mm Hg ECS increased measured perimalleolar subcutaneous pressure in patients with CVI with and without clinical edema but not in control patients. These results suggest the mechanism of benefit of ECS in patients with CVI is due at least in part to an increase in subcutaneous pressure that may act to promote more efficient absorption of perimalleolar extracellular fluid. PMID- 8230565 TI - Foot volumetry can predict recurrent ulceration after subfascial ligation of perforators and saphenous ligation. AB - PURPOSE: The purpose of this study was to determine whether foot volumetry is of value in predicting the outcome after superficial venous surgery for chronic venous ulceration. METHODS: Fifty-three patients who underwent subfascial ligation of perforating veins with or without saphenous vein ligation for recurrent venous ulceration had foot volumetry performed before operation, immediately after operation (1 month), and at regular intervals during follow-up (median 60, range 3 to 144 months). RESULTS: Fourteen patients (26%) had recurrent ulceration (median time to recurrence 48, range 10 to 72 months) and 39 patients remained ulcer free. Before operation expulsion fraction (EF) and half refilling time (HRT) were significantly higher in the no recurrence group. Both groups gained improvement in EF and HRT from their surgery; however, although this was sustained in the no recurrence group, by the time of first recurrence in the recurrence group, EF and HRT had returned to preoperative levels. CONCLUSIONS: Preoperative foot volumetry can therefore predict failure after subfascial perforator and saphenous vein ligation, probably by identifying patients with deep venous incompetence. After operation deterioration in foot volumetry and can also be used to predict the onset of recurrent ulceration. PMID- 8230566 TI - Does air plethysmography correlate with duplex scanning in patients with chronic venous insufficiency? AB - PURPOSE: Duplex ultrasonography with distal cuff deflation was used to determine the presence and size of incompetent veins and compare the results with those of air plethysmography in patients with chronic venous insufficiency. METHODS: Thirty-two legs underwent a detailed study with both modalities. Sixteen legs had venous ulceration, six had stasis dermatitis, and ten had symptomatic varicose veins without skin changes. RESULTS: Although the venous filling index (VFI) in limbs with ulcers (5.4 +/- 3.8 ml/sec) and dermatitis (7.7 +/- 4.6 ml/sec) was significantly higher (p < 0.05) than it was in limbs with varicose veins (2.6 +/- 1.7 ml/sec), there was a large amount of overlap. Only 13% of ulcerated legs had VFI greater than 10 ml/sec. Sixty-three percent of legs with ulcers, 33% of legs with dermatitis, and 90% of legs with varicose veins had VFIs less than 5 ml/sec. Mean ejection fractions (EFs) in the three groups were similar, ranging from 45% to 52%. Combining VFI and EF did not lessen the overlap between groups. Forty-one percent of limbs with ulcers or dermatitis had air plethysmography parameters in the normal or intermediate area (VFI < 5 ml/sec; EF > 40%), which in previous studies corresponded to an incidence of ulceration of only 2%. VFI had a significant but weak correlation (r = 0.39) with the diameter of incompetent veins at the knee and a somewhat stronger relationship (r = 0.55) with the diameter of lower leg veins. Total venous volume correlated moderately well with calf vein diameter (r = 0.75). The clinical status of the leg did not correlate with the diameters of incompetent veins at the knee or calf levels. All limbs with an obstructed outflow had EFs less than 60% and ulcers or dermatitis. CONCLUSIONS: We conclude that plethysmographic measurements of functional venous parameters (VFI,EF) do not discriminate well between limbs with uncomplicated varicose veins and limbs with ulcers or stasis dermatitis and that VFI correlates poorly with the presence of incompetent veins and their diameters. Both duplex scanning and plethysmography seem to be necessary for a complete evaluation of limbs with chronic venous insufficiency. PMID- 8230567 TI - Can air plethysmography accurately identify upper extremity deep venous thrombosis? AB - PURPOSE: Air plethysmography (APG) is an established, noninvasive means of evaluating lower extremity venous outflow. METHODS: To determine whether APG could identify deep venous thrombosis (DVT) of the upper extremity, we measured the percentage of venous outflow in the first second (maximum venous outflow [MVO]) of four groups of arms: (1) normal volunteers (no previous central vein catheters), (2) the affected, (3) the unaffected arms of the patients with DVT (proven by venography or duplex scanning), and (4) hospital patients (no previous central vein catheters). RESULTS: The results of an analysis of variance were as follows: the mean MVO (%) of the affected arms of patients with DVT, 29.4 +/- 2.6, was significantly less than their unaffected arms, 55.3 +/- 3.9, hospital controls, 64.3 +/- 2.2, and normal volunteers, 64.2 +/- 0.9 (p < 0.05). In addition, the unaffected arms differed significantly from all groups (p < 0.05). An MVO of 45% was 2 SD below the mean MVO of the normal volunteer group. Two unaffected arms of the patients with DVT were below 45%, whereas only one affected arm of the patients with DVT was above 45% (45.6%). Therefore a comparison of the distribution of the MVO values revealed virtually no overlap of the affected arms of the patients with DVT with the other three groups. CONCLUSIONS: We conclude that APG clearly discriminated arms with documented DVT from controls and is a highly accurate means of identifying upper extremity DVT. PMID- 8230568 TI - Analysis of the connective tissue matrix and proteolytic activity of primary varicose veins. AB - PURPOSE: Valvular incompetence and venous wall abnormalities have been suggested as primary etiologic factors responsible for the development of varicose veins. This study was conducted to evaluate the connective tissue constituents of greater saphenous varicosities. Proteolytic activity, a factor that can lead to matrix degradation and cause weakening and dilation of the venous wall, was also assessed. METHODS: The collagen and elastin contents of 16 nonthrombophlebitic greater saphenous varicose veins (VV) and seven normal greater saphenous veins (NV) were quantified. In addition, four duplex scanning-confirmed competent segments of greater saphenous veins (i.e., potential varicose veins [PV]) affected by varicosis at alternate sites were analyzed. Proteolytic activity was determined by zymography and radiolabeled substrate assay. RESULTS: The content of collagen was significantly increased in the VV and PV compared with NV (VV = 189 +/- 7 mg/gm, PV = 189 +/- 9 mg/gm vs NV = 144 +/- 10 mg/gm, p < 0.05). Conversely, the elastin content in the VV and PV was significantly reduced (VV = 53 +/- 3 mg/gm, PV = 50 +/- 4 mg/gm vs NV = 74 +/- 4 mg/gm, p < 0.05). The collagen to elastin ratio demonstrated an alteration in VV and PV compared with NV (VV = 3.7 +/- 0.3, PV = 3.9 +/- 0.4 vs NV = 2.0 +/- 0.2, p < 0.05). Casein and gelatin zymography did not demonstrate significant qualitative differences in the enzymatic activities among the three groups. Quantitative analysis of the elastase activity in the venous tissues was similarly not appreciably altered (VV = 5.1 +/- 0.2 U/gm, PV = 5.3 +/- 0.2 U/gm vs NV = 5.7 +/- 0.3 U/gm). CONCLUSION: A significant increase in the collagen content and a significant reduction in the elastin content of VV were demonstrated. The net increase in the collagen/elastin ratio is indicative of an imbalance in the connective tissue matrix. The biochemical profile of PV was similar to VV and significantly different from NV. These preliminary data support the presence of connective tissue abnormalities before valvular insufficiency. In addition, the absence of an increase in the proteolytic activity excludes enzymatic matrix degradation as an essential component in the formation of venous varicosities. PMID- 8230569 TI - Outpatient duplex scanning for deep vein thrombosis: parameters predictive of a negative study result. AB - PURPOSE: Although clinical examination has been reported to be unreliable in the diagnosis of deep vein thrombosis (DVT), this conclusion has often been derived from hospitalized patients (HP) and may not be applicable in an outpatient setting. This study was undertaken to define clinical parameters that might predict the diagnostic value of emergency venous duplex scanning (VDS). METHODS: Venous duplex scans performed over a 9-month period (interval I) in 154 outpatients (154 limbs) and 145 HP (145 limbs) with suspected DVT were reviewed. RESULTS: Eighteen percent of scans of outpatients and 31% of scans of HP were interpreted as positive for lower extremity DVT. With stepwise logistic regression analysis, criteria predictive of a negative result of outpatient VDS included (1) duration of symptoms greater than 7 days (p = 0.003), (2) thigh circumference difference relative to the uninvolved side of less than 3 cm (p = 0.001), and (3) no history of neoplasia (p = 0.03). This model, when applied prospectively to 68 outpatients (68 limbs) over the next 5 months (interval II), yielded a negative predictive value (NPV) of 96.7% (sensitivity 90.9%, specificity 50.9%, positive predictive value 26.3%). Of the 222 outpatients examined during intervals I and II, 98 (44%) met these three clinical criteria. Three of these 98 outpatients had DVT on VDS and thus would have been misclassified as having a negative result. With a similar logistic regression analysis for HP, clinical criteria achieved an NPV of only 75% (sensitivity 36%, specificity 90%, positive predictive value 62%). CONCLUSION: Clinical assessment is unreliable in the diagnostic evaluation of HP with suspected DVT. In an outpatient population, however, clinical evaluation with the above criteria achieved an NPV of 96.7% in the diagnosis of DVT. These parameters may be useful as guidelines in determining the appropriateness of emergency outpatient VDS. PMID- 8230570 TI - Compliance of vascular anastomoses with polybutester and polypropylene sutures. AB - PURPOSE: Polybutester suture is more easily stretched than other vascular sutures and may produce more compliant anastomoses. The effects of using polybutester and polypropylene sutures were compared acutely in arterial autografts and in chronic implants of cephalic vein grafts into the femoral arteries of dogs. METHODS: Paraanastomotic profiles of diameter and compliance were measured with echo tracked ultrasonography, and profiles of intimal thickening were generated from histologic sections of the vessels harvested after 3 months. RESULTS: Polybutester produced more compliant anastomoses, compared with polypropylene, in arterial autografts (in vitro: 5.9% +/- 2.0% vs 3.3% +/- 0.6% diameter change/100 mm Hg, p < 0.01; in vivo: 3.1% +/- 1.1% vs 1.6% +/- 0.5%, p < 0.05), but this difference was not observed with vein as the graft material, either initially (1.1% +/- 1.2% vs 1.7% +/- 0.5%) or after 3 months (2.1% +/- 1.2% vs 2.4% +/- 0.8%). This dichotomy may reflect a governing influence of the stiffer veingrafts, compared with host artery (2.6% +/- 1.0% vs 5.4% +/- 1.2%), or the use of suboptimal tension on the polybutester suture when creating the anastomosis. CONCLUSIONS: Both sutures produced similar compliance and thickness profiles. Polybutester initially produces a more compliant anastomosis when both artery and graft are compliant, reducing anastomotic compliance mismatch. However, this benefit may not apply when the anastomosis includes a vessel of low compliance. PMID- 8230571 TI - An assessment of the long saphenous vein for potential use as a vascular conduit after varicose vein surgery. AB - PURPOSE: There is controversy in the surgical management of varicose veins between stripping of the long saphenous vein (LSV) and high ligation. Moreover, preservation of the LSV is desirable for future coronary or peripheral artery bypass. We have studied 75 limbs in 44 patients after high saphenous ligation with multiple stab phlebectomy. METHODS: Subjective assessment of the outcome of surgery was made with a linear analog scale, and objective cosmetic outcome was assessed by an independent observer (LF) who had not been involved in the surgical treatment of these patients with our modification from the criteria first described by Jakobsen. Patency, length, and diameter of the LSV was measured 6 to 14 months (mean 12 months) after operation with a duplex scanner and a color-flow scanner. Valvular incompetence in the LSV and perforators was also assessed. RESULTS: Results show a good subjective and objective outcome in 95% and 97% of limbs, respectively. The LSVs were patent from ankle to groin in 68% and from ankle to knee in 82%, with a mean diameter of 4.0 +/- 0.1 mm (mean +/- SEM). There was no statistically significant difference in symptomatic outcome and presence of reflux in the LSV (X2 = 0.465; p = 0.4954; NS) or objective cosmetic outcome and the presence of reflux in the LSV (chi 2 = 2.916; p = 0.0877; NS). CONCLUSIONS: It is concluded that high saphenofemoral ligation combined with multiple "stab avulsions" preserves an LSV with characteristics suggesting suitability for future use as a vascular prosthesis with good early symptomatic and cosmetic results. PMID- 8230572 TI - Percutaneous transluminal renal angioplasty versus surgical reconstruction of atherosclerotic renal artery stenosis: a prospective randomized study. AB - PURPOSE: The purpose of this prospective randomized study was to compare percutaneous transluminal renal angioplasty (PTRA) and operation as initial therapy with regard to technical results, primary and secondary patency, and effects on blood pressure and renal function in patients with atherosclerotic unilateral renal artery stenosis. METHODS: Fifty-eight patients who did not have diabetes, who were less than 70 years of age, and who had severe hypertension and significant stenosis were randomized to receive PTRA or operation. Angiography was performed 10 days, 1 year, and 2 years after treatment to verify patency, and blood pressure and renal function were simultaneously evaluated. RESULTS: Technically, PTRA was successful in 83% and operation in 97% of patients. The primary patency rate after 24 months was 75% in the PTRA group and 96% in the operative group in technically successful cases. The secondary patency rate in the PTRA group was 90% and in the surgical group 97%. To achieve these results four patients in the PTRA group required operation, and one patient in the surgical group required PTRA. Hypertension was cured or improved after additional treatment in 90% of the patients after PTRA and 86% after operation. The corresponding figures for improved or unchanged renal function were 83% and 72%, respectively. After additional treatment, effects on blood pressure and renal function did not differ. Seventeen percent of the patients treated with PTRA required surgical intervention. CONCLUSIONS: PTRA is recommended as first choice of therapy for atherosclerotic renal artery stenosis causing renovascular hypertension if combined with intensive follow-up and aggressive reintervention. PMID- 8230573 TI - Endotoxemia after major vascular operations. AB - PURPOSE: Endotoxemia has been associated with syndromes such as adult respiratory distress syndrome and multiple organ failure. Translocation of endotoxins from the gut has been demonstrated to occur experimentally in animals after splanchnic ischemia. METHODS: We investigated endotoxemia in 11 patients with hemorrhagic shock caused by a ruptured abdominal aortic aneurysm and in five patients after elective abdominal aortic aneurysm repair. Endotoxin was measured quantitatively with a limulus assay with a detection limit of 12.5 pg/ml. RESULTS: In 7 of the 11 patients admitted to the hospital with an acute condition endotoxin was present on admission (range 15 to 25 pg/ml), even before resuscitation and operation were started. In patients treated electively endotoxin was noticed after cross-clamping of the aorta in four of five patients (mean +/- SEM: 18.9 +/ 4.6 pg/ml). After reperfusion endotoxemia increased and was present in all patients (22.8 +/- 2.8 pg/ml). All five patients treated electively had an uneventful recovery without specific organ failures or infectious complications. CONCLUSIONS: It is concluded that systemic endotoxemia as monitored by the limulus assay is a common finding in patients after major vascular operation. Hemorrhagic shock resulting in splanchnic ischemia may lead to endotoxemia. Ischemia and especially reperfusion after aortic cross-clamping also result in endotoxin translocation. However, the low concentrations of systemic circulating endotoxin found were not related to subsequent adverse effects of either the patients treated for acute conditions or in the electively treated patients. PMID- 8230574 TI - The preservation of renal function by isovolemic hemodilution during aortic operations. AB - PURPOSE: In an investigation of the effects of isovolemic hemodilution, 39 consecutive patients undergoing elective infrarenal aortic operation had detailed measurements of renal function, renal artery blood flow, and cardiac hemodynamics. METHODS: The patients were randomly allocated to receive acute preoperative isovolemic hemodilution to a hematocrit of 28%, with 20 patients receiving hemodilution and 19 being control subjects. RESULTS: Twelve (63%) of the control group had renal impairment, compared with only four (20% in the group receiving hemodilution (p < 0.01). Hemodilution also prevented the fall in cardiac output induced by cross-clamping (p < 0.01) and significantly reduced the need for transfusion of donor blood (p < 0.01). CONCLUSIONS: Acute isovolemic hemodilution is clearly a useful adjunct in the management of patients undergoing elective aortic operation. PMID- 8230575 TI - Tibial bypass grafting for limb salvage with ringed polytetrafluoroethylene prostheses: results of primary and secondary procedures. AB - PURPOSE: In advanced peripheral ischemia, tibial artery prosthetic bypass grafting is virtually the last chance for limb salvage when conservative treatment and other methods of revascularization have failed. METHODS: For this study we reviewed our 7 years of experience with below-popliteal artery polytetrafluoroethylene grafts implanted for limb salvage. RESULTS: All results regarding graft patency and limb salvage were computed by actuarial methods and were presented in the form of life-table analysis. There were 211 grafts in 184 patients (195 limbs). Two and 5 years after the operation, primary patency rates were 37% and 23% and secondary patency rates were 45% and 25%, respectively. The main negative predictive factors for patency rate were an occluded primary plantar arch artery and poor vessel runoff. Primary bypass procedures had markedly better success rates than secondary (repeat) procedures, with primary patency rates of 52% at 2 years and 42% at 4 years, whereas the corresponding rates for secondary procedures were 22% and 14%, respectively. The 5-year cumulative limb salvage rate was 51% for the total series. CONCLUSIONS: Although the patency of prosthetic grafts is not as satisfactory as that of autologous vein grafts, the limb salvage rate justifies an aggressive approach to infrapopliteal bypass grafting with polytetrafluoroethylene prostheses when the limb is threatened and vein is not available. PMID- 8230576 TI - Limitations of peroneal artery bypass grafting for limb salvage. AB - PURPOSE: The purpose of this study was to compare the results of peroneal bypass grafting for limb salvage with the results of other tibial and pedal bypass grafts performed concurrently. METHODS: Thirty-four peroneal bypass grafts with autologous vein were performed for limb salvage between September 1986 and June 1992. These constituted 18% of an overall experience of 194 tibial or pedal bypasses performed during that time. Preoperative and intraoperative arteriograms were reviewed to identify anatomic characteristics associated with successful limb salvage. RESULTS: Secondary patency rates for peroneal bypass grafts (70%) compared with the other tibial and pedal bypass grafts (65%) did not differ significantly at 48 months by life-table analysis. Limb salvage achieved by peroneal artery bypass grafting was significantly worse (55%) than that achieved by the remaining tibial and pedal bypasses (67%) at 48 months. Limb salvage was 33% at 7 months for those undergoing peroneal artery bypass grafting as opposed to 57% at 48 months for patients undergoing other tibial or pedal revascularizations with tissue necrosis. Four anatomic features were identified that were associated with failure after peroneal artery bypass grafting. These were peroneal length less than 10 cm (p = 0.012), peroneal artery diameter less than 2 mm (p = 0.035), absence of arteriographically demonstrated collaterals perfusing the foot (p = 0.0001), and little or no visualization of the pedal arch (p = 0.008). CONCLUSIONS: Although successful grafts may avoid amputation in carefully selected cases, alternatives to peroneal artery bypass grafting should be considered when less than favorable anatomic conditions are encountered, particularly in the presence of forefoot tissue necrosis. PMID- 8230577 TI - Successful management of traumatic false aneurysm of the extracranial vertebral artery by duplex-directed manual occlusion: a case report. AB - A 32-year-old man was transferred to our hospital after a 2.0 by 2.5 cm traumatic false aneurysm of the distal extracranial vertebral artery was noted after a stab wound of the posterior side of the neck. To obviate the need for operative exposure of the distal vertebral artery at the base of the skull, we elected to perform duplex-directed manual occlusion of the lesion. Angiography before and after the procedure, as well as 10-month follow-up duplex ultrasonography, demonstrated satisfactory thrombosis of the false aneurysm without evidence of a residual arterial defect. There was no morbidity associated with the procedure. We conclude that duplex-directed manual occlusion, a new technique recently described for the nonoperative management of postcatheterization femoral false aneurysms, can be applied safely and effectively to false aneurysms in other locations in which the risks and technical difficulties of operative repair render surgery less desirable. PMID- 8230578 TI - Further developments concerning practice expense reimbursement in vascular surgery. PMID- 8230579 TI - X-linked ichthyosis. PMID- 8230580 TI - Variety of roles for molecular genetics in dealing with cardiovascular disease. PMID- 8230581 TI - The genetic revolution: despite perfection of elegant techniques, ethical answers still elusive. PMID- 8230582 TI - First immunotoxin therapy for many common solid tumors enters phase I clinical trial. PMID- 8230583 TI - From the Food and Drug Administration. PMID- 8230584 TI - From the Centers for Disease Control and Prevention. Update: hantavirus pulmonary syndrome--United States, 1993. PMID- 8230585 TI - Oral contraceptive use and infectivity of HIV-seropositive women. PMID- 8230586 TI - 'Technology follies': curtain call. PMID- 8230587 TI - Thermocautery causes a gauze pad fire. PMID- 8230588 TI - Changes in bone density with lactation. PMID- 8230589 TI - Seabather's eruption or diver's dermatitis? PMID- 8230590 TI - Loperamide to diagnose Cushing's syndrome. PMID- 8230591 TI - Colorectal cancer. PMID- 8230592 TI - Tay-Sachs disease--carrier screening, prenatal diagnosis, and the molecular era. An international perspective, 1970 to 1993. The International TSD Data Collection Network. AB - OBJECTIVES: To provide an update of the international experience with carrier screening and prenatal diagnosis for Tay-Sachs disease (TSD), to assess the impact of these efforts, and to review the recent developments in DNA technology with application to TSD carrier detection and screening. DESIGN: Through the International TSD Testing, Quality Control, and Data Collection Center, all testing centers in the world were surveyed annually to assess overall experience with carrier testing and prenatal diagnosis. Quality control and laboratory surveillance of testing centers were performed through an annual assessment, using samples provided by the center. SETTING: Tay-Sachs disease testing centers around the world. PARTICIPANTS: Nearly 1 million young adults from both Jewish and non-Jewish populations. INTERVENTION: Gene product screening (enzyme testing) and DNA-based mutation analysis (in some populations). MAIN OUTCOME MEASURE: Impact of screening program on disease incidence. RESULTS: Data from all centers in the international TSD network on experience with TSD carrier testing and prenatal diagnosis since 1974 indicated that more than 36,000 heterozygotes were identified and 1056 couples found to be at risk for TSD in their offspring. A total of 2416 pregnancies at increased risk for TSD were monitored by amniocentesis or chorionic villus sampling. A dramatic decrease in the incidence of TSD in the Jewish populations was demonstrated. With both serum and leukocyte proficiency testing, there have been only 16 instances (of 845 cumulative laboratory evaluations) of one or more errors reported by a laboratory since 1983 resulting in nonaccreditation. CONCLUSIONS: This analysis represents a prototypic effort in coordinating adult education, carrier testing, and genetic counseling directed toward prospective prevention of a uniformly fatal childhood disease and demonstrates that such an effort can dramatically affect disease incidence. PMID- 8230593 TI - DNA diagnosis of neurofibromatosis 2. Altered coding sequence of the merlin tumor suppressor in an extended pedigree. AB - OBJECTIVE: To define the DNA mutation causing neurofibromatosis 2 (NF2), a severe genetic disorder involving the development of multiple nervous system tumors in adulthood, in a large, well-studied NF2 pedigree previously used to chromosomally map and to isolate the disease gene. DESIGN: Single-strand conformational polymorphism (SSCP) and DNA sequence analysis of the NF2 gene amplified from affected and unaffected family members. PARTICIPANTS: Affected, unaffected, and at-risk members of a large pedigree segregating NF2, an autosomal dominant disorder caused by inactivation of the merlin tumor suppressor encoded in chromosome band 22q12. RESULTS: A DNA alteration in the merlin coding sequence caused a shift on SSCP gels that was characteristic of the disease chromosome in this NF2 pedigree, being transmitted with the disorder, present only in affected members of the pedigree, absent in unaffected members of the family, and absent from 158 unrelated individuals. The alteration caused substitution of a tyrosine for an asparagine at position 220 of the merlin protein, in a region highly conserved in closely related members of the family of cytoskeletal-associated proteins. The DNA change could also be detected by restriction enzyme digestion with Rsa I. CONCLUSION: Current practice dictates screening of all those "at risk" for NF2 with magnetic resonance imaging, but the frequency and duration of screening are problematic because of the variable course of the disease. The identification of a DNA alteration in the NF2 gene will permit predictive molecular testing of individuals at risk in this specific family, sparing the expense and emotional burden of protracted screening programs. This information, by providing diagnostic certainty, should also reduce psychological and financial burdens and improve medical care for affected family members. A similar approach to defining the underlying lesion and developing a predictive test is applicable in any documented NF2 family. PMID- 8230594 TI - Attitudes toward direct predictive testing for the Huntington disease gene. Relevance for other adult-onset disorders. The Canadian Collaborative Group on Predictive Testing for Huntington Disease. AB - OBJECTIVE: To assess attitudes toward, and projected utilization of, direct mutation testing by individuals at risk for Huntington disease (HD). DESIGN: Prior to the cloning of the gene for HD, a questionnaire concerning the use of a definitive test was constructed and mailed to 354 participants in the Canadian Collaborative Study for HD. Completed questionnaires were received from 250 participants (response rate, 71%). Persons were asked to indicate whether they would participate in a new predictive test that was either 100% accurate (the definitive test, requiring blood only from the proband) or only 99% accurate. RESULTS: Most (72%) of the persons who had previously received a result in a predictive testing program said they would request testing in either situation. Significantly more persons would request the definitive test than the 99% accurate test (72% vs 58%; P < .02). Respondents for whom testing was uninformative in the linkage test program or who had previously received an increased-risk result were more likely to indicate they would use the test than those who received a decreased-risk result or chose not to have the original test (P = .0003). Less than half (46%) of the participants who initially chose not to have the linkage test said they would return for the new direct test. The major factor that has limited acceptance of predictive testing for this group is the concern about receiving an increased-risk result in the absence of any therapy to alter progression of the disease. CONCLUSIONS: A direct mutation test for HD will most readily be accepted by persons who wanted but could not previously receive a result in the linkage test program and those who previously received an increased risk result. In the absence of therapy, the majority of persons who previously chose not to have predictive testing are unlikely to participate in a new test despite improved accuracy. This has implications for the expected demands for testing services for other adult-onset genetic disorders. PMID- 8230595 TI - Inherited primary peripheral neuropathies. Molecular genetics and clinical implications of CMT1A and HNPP. PMID- 8230596 TI - The clinical applications of new DNA diagnostic technology on the management of cancer patients. PMID- 8230597 TI - Progress toward human gene therapy. PMID- 8230598 TI - Privacy rules for DNA databanks. Protecting coded 'future diaries'. AB - In privacy terms, genetic information is like medical information. But the information contained in the DNA molecule itself is more sensitive because it contains an individual's probabilistic "future diary," is written in a code that has only partially been broken, and contains information about an individual's parents, siblings, and children. Current rules for protecting the privacy of medical information cannot protect either genetic information or identifiable DNA samples stored in DNA databanks. A review of the legal and public policy rationales for protecting genetic privacy suggests that specific enforceable privacy rules for DNA databanks are needed. Four preliminary rules are proposed to govern the creation of DNA databanks, the collection of DNA samples for storage, limits on the use of information derived from the samples, and continuing obligations to those whose DNA samples are in the databanks. PMID- 8230599 TI - Medical genetics. A 40-year perspective on the evolution of a medical specialty from a basic science. PMID- 8230600 TI - Isolating fetal cells from maternal blood. Advances in prenatal diagnosis through molecular technology. AB - OBJECTIVES: To review the rationale for and progress toward the goal of isolating and analyzing fetal cells circulating in maternal blood, and to explore the feasibility of this method in providing noninvasive prenatal cytogenetic diagnosis. DATA SOURCES: Critical review of data published since the first report (1969) of fetal metaphases in maternal blood. Emphasis is placed on data since the demonstration by polymerase chain reaction (PCR) in 1989 and 1990 that fetal cells indeed exist in maternal blood. DATA SYNTHESIS: Clinical evaluations have not yet been conducted, but it is already clear that molecular technologies have allowed the unequivocal demonstration of fetal cells in maternal blood. Using PCR, our own group and others have demonstrated Y sequences and single gene sequences (eg, hemoglobin LeporeBoston) in maternal blood. Thus, fetal DNA sequences indeed exist in maternal blood. Among the various candidate cells, the most promising appear to be fetal nucleated red blood cells. We isolated nucleated red blood cells on the basis of flow-sorting for the transferrin receptor and glycophorin-A. Enriched samples were then subjected to fluorescence in situ hybridization with chromosome-specific probes. This approach allowed us to detect trisomy 21 and trisomy 18, work later confirmed by others. CONCLUSIONS: Isolating and analyzing fetal cells from maternal blood is clearly possible. Several key biologic questions remain--the optimal cells for isolation, frequency of cells in maternal blood, timing during gestation for maternal blood sampling, and the likelihood of persistence of fetal cells after delivery. Clinical evaluations planned by the National Institute of Child Health and Human Development will determine the sensitivity and specificity of this method and its precise role in prenatal cytogenetic diagnosis. PMID- 8230601 TI - Genetic factors in multiple sclerosis. AB - OBJECTIVE: To evaluate the role of candidate genes in the susceptibility to multiple sclerosis (MS) and describe the role of T-cell receptor (TCR) gene rearrangements in the MS brain lesion in identifying a major target of the immune response in this disease. DATA SOURCES: MEDLINE, bibliography review of published data, and unpublished studies. STUDY SELECTION: Published studies using novel molecular approaches to analyze the role of the major histocompatibility complex (MHC) and TCR gene complexes, as well as other candidate genes, in susceptibility to MS. We analyze epigenetic events involving TCR genes in individuals with MS and describe recent clinical trials in which immunotherapy has been attempted. DATA SYNTHESIS: Consistent with a polygenic model for disease predisposition, MHC and TCR gene associations with MS are relatively weak. Despite intensive research, no other putative "MS genes" have been firmly established. The analysis of TCR rearrangements in the brain lesion has helped to identify a major target of the immune response in MS. CONCLUSION: Understanding the genetic basis for autoimmune demyelination will offer new possibilities for the treatment of this illness. PMID- 8230602 TI - The 'pennies drop'. Genetic discoveries of medical significance. PMID- 8230603 TI - Researchers Lupski & Chance study a baffling genetic disease--their own. PMID- 8230604 TI - Hunting dino DNA--Mary Schweitzer puts T rex bones under the microscope. PMID- 8230605 TI - A piece of my mind. The hymn. PMID- 8230607 TI - George L. Fite, MD: a decade at JAMA. PMID- 8230606 TI - Science reporters hear wide range of recent data at 12th annual conference. PMID- 8230608 TI - From the Centers for Disease Control and Prevention. Measles--United States, first 26 weeks, 1993. PMID- 8230609 TI - From the Centers for Disease Control and Prevention. Pregnancy complications and perinatal outcomes among women with diabetes--North Carolina, 1989-1990. PMID- 8230610 TI - Patient decision making. PMID- 8230611 TI - Primary care as part of US Health Services Reform. PMID- 8230612 TI - Primary care as part of US Health Services Reform. PMID- 8230613 TI - Primary care as part of US Health Services Reform. PMID- 8230614 TI - Primary care as part of US Health Services Reform. PMID- 8230615 TI - Primary care as part of US Health Services Reform. PMID- 8230616 TI - Emphasizing chest pain as a symptom of heart attack. PMID- 8230617 TI - AIDS policy: two divisive issues. PMID- 8230618 TI - Disease-specific survival following routine prostate cancer screening by digital rectal examination: corrected patient classification. PMID- 8230619 TI - Resuscitation in near drowning. PMID- 8230620 TI - Psychological predictors of hypertension in the Framingham Study. Is there tension in hypertension? AB - OBJECTIVE: To test the hypothesis that heightened anxiety, heightened anger intensity, and suppressed expression of anger increase the risk of hypertension, using the Framingham Heart Study. DESIGN: A cohort of men and women without evidence of hypertension at baseline were followed up for 18 to 20 years. Baseline measures of anxiety (tension), anger symptoms, and expression of anger (anger-in and anger-out) were taken, along with biological and behavioral predictors of hypertension (initial systolic blood pressure, heart rate, relative weight, age, hematocrit, alcohol intake, smoking, education, and glucose intolerance). PARTICIPANTS: A total of 1123 initially normotensive persons (497 men, 626 women) were included. Analyses were stratified by age (45 to 59 or > or = 60 years) and gender. MAIN OUTCOME MEASURES: Hypertension was defined as either taking medication for hypertension or blood pressures higher than 160/95 mm Hg at a biennial examination. RESULTS: In univariate analyses, middle-aged men who went on to develop hypertension had greater baseline anxiety levels than men who remained normotensive (P = .04). Older hypertensive men had fewer anger symptoms at baseline (P = .04) and were less likely to hold their anger in (P = .01) than normotensives. In multivariate Cox regression analysis including biological predictors, anxiety remained an independent predictor of hypertension in middle aged men (P = .02). Among older men, anger symptoms and anger-in did not remain significant predictors in the multivariate analysis. Further analysis showed that only middle-aged men with very high levels of anxiety were at increased risk (relative risk, 2.19; 95% confidence interval, 1.22 to 3.94). No psychological variable predicted hypertension in middle-aged or older women in either univariate or multivariate analyses. CONCLUSIONS: The results indicate that among middle-aged men, but not women, anxiety levels are predictive of later incidence of hypertension. PMID- 8230621 TI - Positive predictive value of screening mammography by age and family history of breast cancer. AB - OBJECTIVE: To determine the positive predictive value (PPV) of low-cost screening mammography according to age and family history of breast cancer. DESIGN: Cross sectional. SETTING: Six counties in northern California. PARTICIPANTS: A total of 31,814 women aged 30 years and older referred for mammography to the University of California, San Francisco, Mobile Mammography Screening Program from April 18, 1985, through November 20, 1992. MEASUREMENTS: Breast cancer risk profile, two standard mammographic views per breast, and follow-up of abnormal screening examinations. RESULTS: Although women aged 50 years or older constituted only 38.3% of all women who received first-screening mammography, 74% of breast cancers were detected in this group. Ten cancers were diagnosed per 1000 first screening examinations in women aged 50 years or older, with 14.8 diagnostic procedures per cancer diagnosed compared with two cancers per 1000 screening examinations and 48.3 diagnostic tests per cancer diagnosed in women younger than 50 years. The PPV of first-screening mammography (number of breast cancers detected per abnormal examination) increased with age: .03 for those aged 30 to 39 years; .04 for those aged 40 to 49; .09 for those aged 50 to 59; .17 for those aged 60 to 69; and .19 for those aged 70 years or older (chi 2 for trend, P < .001). Women aged 50 to 59 years had a higher PPV for first-screening mammography than women aged 40 to 49 years (.09 vs. .04; P = .004), and women with a family history of breast cancer had higher PPVs compared with women without history (40 to 49 years of age, .13 vs .04, P = .01; and 50 to 59 years of age, .22 vs .09, P = .01). CONCLUSION: Five times as many cancers per 1000 first-screening mammographic examinations were diagnosed in women aged 50 years or older compared with women aged less than 50 years. The highest PPVs for mammography were in women aged 50 years or older and in women aged 40 years or older with a family history of breast cancer. Efforts to promote screening mammography should focus on women in these groups, in whom the majority of breast cancers occur and for whom mammography has the highest PPVs. PMID- 8230622 TI - Mortality following ventricular arrhythmia suppression by encainide, flecainide, and moricizine after myocardial infarction. The original design concept of the Cardiac Arrhythmia Suppression Trial (CAST). AB - OBJECTIVE: To test the hypothesis that in survivors of myocardial infarction, the suppression of ventricular premature depolarizations improves survival free of cardiac arrest and arrhythmic death. DESIGN: International, prospective, multicenter, randomized, placebo-controlled trial. SETTING: University and community hospitals. PATIENTS: A total of 3549 patients with myocardial infarction and left ventricular dysfunction. INTERVENTION: Administration of encainide, flecainide, moricizine, or placebo to suppress ventricular premature depolarizations. MAIN OUTCOME MEASURES: Overall survival and survival free of cardiac arrest or arrhythmic death were compared in patients randomized to long term, active antiarrhythmic drug therapy vs corresponding placebo, using the stratified log rank statistic. RESULTS: At 1 year from the time of randomization to blinded therapy, 95% of placebo-treated patients vs 90% of active drug-treated patients remained alive (P = .0006). Similarly, at 1 year, 96% of placebo-treated patients vs 93% of active drug-treated patients remained free of cardiac arrest or arrhythmic death (P = .003). CONCLUSIONS: The suppression of asymptomatic or mildly symptomatic ventricular arrhythmias after myocardial infarction does not improve survival and can increase mortality. Treatment strategies designed solely to suppress these arrhythmias should no longer be followed. PMID- 8230623 TI - Clinical experience with penicillin skin testing in a large inner-city STD clinic. AB - OBJECTIVE: To establish (1) the prevalence of positive penicillin skin tests among outpatients with well-defined but variable history of penicillin allergy and (2) the reproducibility, safety, and negative predictive value of skin testing with benzylpenicilloyl polylysine (PPL) and a minor-determinant mixture (MDM). DESIGN: Serial consenting outpatients with current indications for penicillin therapy were skin-tested in duplicate with PPL and MDM. Subjects with negative skin tests (93% of those positive by history and 95% of those negative by history) received therapeutic courses of benzylpenicillin (81%) or ampicillin (19%). Negative predictive value of skin testing was established by 72-hour follow-up for adverse reactions to drug. SETTING/PATIENTS: A total of 5063 consecutive, qualifying outpatients in a Baltimore, Md, sexually transmitted disease (STD) clinic. The study group was young (73% between 20 and 40 years old), 66% male, and 90% black; 25% had history of atopy. Follow-up was 94% complete. RESULTS: Positive skin tests were observed in 7.1% of 776 individuals with previous history of penicillin allergy and in 1.7% of 4287 subjects negative by history (P << .001). Previous history of anaphylaxis or urticaria was associated with significantly higher rates of positive skin tests of 17.3% and 12.4%, respectively (P<<.001). Only 4% with history of exanthem had positive skin tests (P = .03). The coefficient of variation for duplicate skin tests was 11%. Time intervals since last penicillin treatment did not influence the rate of positive skin tests. Adverse reactions to skin tests occurred in 13 (1.2% of patients positive by history; 9.4% of those with positive skin tests). A mild anaphylactic reaction occurred in one individual whose preliminary scratch testing was inadvertently omitted; systemic pruritus or urticaria occurred in 11 subjects; one had a large local reaction. After penicillin administration to individuals with negative skin tests, acute allergic reactions occurred in 0.5% of subjects negative by history compared with 2.9% of subjects positive by history (chi 2 = 33.3; P = .0001). Reactions were generally mild and self limited; only two cases of mild anaphylactic reaction occurred, both in patients with history of severe IgE-mediated reaction. CONCLUSIONS: Skin testing with both major and minor penicillin determinants is safe using current recommendations, and both reagents are necessary for maximizing the identification of sensitized subjects. Routine penicillin skin testing can facilitate the safe use of penicillin in 90% of individuals with a previous history of penicillin allergy. PMID- 8230624 TI - Rationing failure. The ethical lessons of the retransplantation of scarce vital organs. AB - Because of a shortage of transplantable livers and hearts, the transplant community has had to decide--by who gets an organ--who lives or dies. Despite this shortage, whether one has previously received a transplant is not used as a criterion to distribute organs. The existing allocation system distributes 10% to 20% of available hearts and livers to retransplant patients. This article examines three differences between primary transplantation and retransplantation that may affect the priority that retransplant candidates should receive in vying for organs: (1) the special obligations that transplant teams have not to abandon patients on whom they have already performed a transplant, (2) the fairness of allowing individuals to get multiple transplants while some die awaiting their first, and (3) the difference in efficacy between primary transplantation and retransplantation. Only this last difference holds up to critical analysis. Our moral duty to direct scarce, lifesaving resources to those likely to benefit from them, suggests that, all other things equal, primary transplant candidates should receive priority because their mortality after transplantation is lower. Consistency also demands that previous transplant history be taken into account, as we already allocate organs according to ABO blood group matching, a factor that affects transplant outcome approximately the same amount as a previous transplantation. We therefore conclude that the system should be revised so that primary transplant candidates have a better chance of receiving organs than retransplant candidates. PMID- 8230625 TI - Continuous intrathecal baclofen infusion for spasticity of cerebral origin. AB - OBJECTIVE: To determine if continuous intrathecal baclofen infusion (CIBI) would provide continuous relief of spasticity in patients with spasticity of cerebral origin, especially children with cerebral palsy. DESIGN: Prospective, unblinded trial, before and after CIBI. SETTING: Children's Hospital of Pittsburgh (Pa). PATIENTS: Thirty-seven patients, 5 to 27 years of age, with spasticity of cerebral origin. INTERVENTION: Continuous intrathecal baclofen infusion for 3 to 48 months. MAIN OUTCOME MEASURES: Muscle tone, range of motion, upper extremity timed tasks, activities of daily living (ADLs). RESULTS: Six and 12 months after CIBI, muscle tone was significantly decreased in the upper (P = .04) and lower (P = .001) extremities. There was a significant relationship between baclofen dosage and muscle tone in the upper (P = .02) and lower (P = .001) extremities. Hamstring motion, upper extremity function, and ADLs were significantly improved in 25 patients who were capable of self-care. CONCLUSION: Spasticity of cerebral origin can be effectively treated with CIBI. Because baclofen dosages can be titrated for the desired clinical response, CIBI is particularly useful for patients who need some spasticity to stand and ambulate. PMID- 8230626 TI - Mortality Probability Models (MPM II) based on an international cohort of intensive care unit patients. AB - OBJECTIVE: To revise and update models in the Mortality Probability Model (MPM II) system to estimate the probability of hospital mortality among 19,124 intensive care unit (ICU) patients that can be used for quality assessment within and among ICUs. DESIGN AND SETTING: Models developed and validated on consecutive admissions to adult medical and surgical ICUs in 12 countries. PATIENTS: A total of 12,610 patients for model development, 6514 patients for model validation. Patients younger than 18 years and burn, coronary care, and cardiac surgery patients were excluded. OUTCOME MEASURE: Vital status at hospital discharge. RESULTS: The admission model, MPM0, contains 15 readily obtainable variables. In developmental and validation samples it calibrated well (goodness-of-fit tests: P = .623 and P = .327, respectively, where a high P value represents good fit between observed and expected values) and discriminated well (area under the receiver operating characteristic curve = 0.837 and 0.824, respectively). The 24 hour model, MPM24 (developed on 10,357 patients still in the ICU at 24 hours), contains five of the admission variables and eight additional variables easily ascertained at 24 hours. It also calibrated well (P = .764 and P = .231 in the developmental and validation samples, respectively) and discriminated well (area under the receiver operating characteristic curve = 0.844 and 0.836 in the developmental and validation samples, respectively). CONCLUSIONS: Among severity systems for intensive care patients, the MPM0 is the only model available for use at ICU admission. Both MPM0 and MPM24 are useful research tools and provide important clinical information when used alone or together. PMID- 8230627 TI - Tension and hypertension. PMID- 8230628 TI - Think tank helping to get HIV immune-restoration trials off the ground. PMID- 8230629 TI - Federal ruling reins in liberal HIV testing law, consensus still lacking on controversial issue. PMID- 8230630 TI - Military medicine faces at least 4 more months of supporting US, other peacekeepers in Somalia. PMID- 8230631 TI - From the Centers for Disease Control and Prevention, Physician and other health care professional counseling of smokers to quit--United States, 1991. PMID- 8230632 TI - From the Centers for Disease Control and Prevention. National poliomyelitis immunization days--People's Republic of China, 1993. PMID- 8230633 TI - From the Centers for Disease Control and Prevention. Mortality trends for selected smoking-related cancers and breast cancer--United States, 1950-1990. PMID- 8230634 TI - Screening strategies for lead poisoning. PMID- 8230635 TI - Screening strategies for lead poisoning. PMID- 8230636 TI - Screening strategies for lead poisoning. PMID- 8230637 TI - Screening strategies for lead poisoning. PMID- 8230638 TI - Pancreatic extract lipase activity. PMID- 8230639 TI - Reimbursement for unproven therapies: the case of thermography. PMID- 8230640 TI - Planned Parenthood v Casey. PMID- 8230641 TI - Increasing age and survival after bone marrow transplant. PMID- 8230642 TI - Maternal age at first childbirth and risk of low birth weight and preterm delivery in Washington State. AB - OBJECTIVE: To study the consequences of delayed first childbearing in a large, population-based US sample, with separate analysis of women aged 40 years or more and adjustment for socioeconomic factors, smoking, medical and reproductive conditions, and route of delivery. DESIGN AND SETTING: Retrospective survey of Washington State birth certificates from 1984 through 1988. SUBJECTS: First liveborn singleton infants of women aged at least 20 years. Of eligible white infants, all those born to women aged 35 to 39 years (n = 4019) and 40 years or more (n = 410) and a maternal age-stratified random sample of white infants of younger women were studied. All eligible black infants were studied. OUTCOME MEASURES: Low (< 2500 g) and very low (< 1500 g) birth weight and preterm delivery (< 37 weeks of gestation). RESULTS: Adjusted odds ratios for delivering a low-birth-weight white infant increased progressively with each 5-year maternal age group, reaching 2.3 (95% confidence interval, 1.6 to 3.4) for women aged 40 years or more compared with those aged 20 to 24 years. The maternal age effects for very low birth weight and preterm delivery were similar; for each, the odds ratio was 1.8 for the oldest group. No significant maternal age effect was found among births of black infants, but only 127 births to women aged 35 years or more were studied. CONCLUSION: Increasing maternal age at first childbirth is an independent risk factor for low birth weight and preterm delivery of white infants in the United States. PMID- 8230643 TI - Ethics consultants' recommendations for life-prolonging treatment of patients in a persistent vegetative state. AB - OBJECTIVE--Surprisingly little is known about the content of ethics consultants' recommendations. We chose to study this issue using hypothetical persistent vegetative state (PVS) cases. We addressed four questions: What recommendations do ethics consultants give regarding life-prolonging treatment (LPT) in PVS cases? To what degree is there consensus? What factors influence recommendations? Do recommendations conform to established guidelines? DESIGN--Questionnaire survey. Our questionnaire asked subjects what they would recommend for seven hypothetical vignettes involving a PVS patient that varied with respect to advance directives and family wishes. We also questioned subjects about demographic characteristics, ethics consultation experience, and personal preference for LPT in PVS. SUBJECTS--Attendees at an annual meeting of the Society for Bioethics Consultation (n = 154). RESULTS--The response rate was 77%. Eighty-one percent of respondents were ethics committee members and 62% were ethics consultants. There was general agreement among respondents for only one of seven vignettes: in the case of a PVS patient whose advance directive and family agree that LPT be stopped, 93% recommended stopping all LPT. Responses to other vignettes varied considerably. Although patient wishes were an important factor influencing recommendations, none of the respondents adhered invariably to the patient's advance directive. Recommendations were also influenced by family wishes, resource allocation considerations, legal constraints, and personal preference for LPT in PVS. Guidelines we examined were generally too equivocal to be useful for evaluating ethics consultants' recommendations. CONCLUSIONS--The finding of wide variability in ethics consultants' recommendations suggests a need to clarify standards for ethics consultation. PMID- 8230644 TI - Female gender as a risk factor for torsades de pointes associated with cardiovascular drugs. AB - OBJECTIVE: To test the hypothesis that female prevalence is greater than expected among reported cases of torsades de pointes associated with cardiovascular drugs that prolong cardiac repolarization. DATA SOURCES: A MEDLINE search of the English-language literature for the period of 1980 through 1992, using the terms torsade de pointes, polymorphic ventricular tachycardia, atypical ventricular tachycardia, proarrhythmia, and drug-induced ventricular tachycardia, supplemented by pertinent references (dating back to 1964) from the reviewed articles and by personal communications with researchers involved in this field. STUDY SELECTION: Ninety-three articles were identified describing at least one case of polymorphic ventricular tachycardia (with gender specified) associated with quinidine, procainamide hydrochloride, disopyramide, amiodarone, sotalol hydrochloride, bepridil hydrochloride, or prenylamine. A total of 332 patients were included in the analysis following application of prospectively defined criteria (eg, corrected QT [QTc] interval of 0.45 second or greater while receiving drug). DATA EXTRACTION: Clinical and electrocardiographic descriptors were extracted for analysis. Expected female prevalence for torsades de pointes associated with quinidine, procainamide, disopyramide, and aminodarone was conservatively estimated from gender-specific data reported for antiarrhythmic drug prescriptions in 1986, as derived from the National Disease and Therapeutic Index, a large pharmaceutical database; expected female prevalence for torsades de pointes associated with sotalol, bepridil, and prenylamine was assumed to be 50% or less since these agents are prescribed for male-predominant cardiovascular conditions. RESULTS: Women made up 70% (95% confidence interval, 64% to 75%) of the 332 reported cases of cardiovascular-drug-related torsades de pointes, and a female prevalence exceeding 50% was observed in 20 (83%) of 24 studies having at least four included cases. When analyzed according to various descriptors, women still constituted the majority (range, 51% to 94% of torsades de pointes cases), irrespective of the presence or absence of underlying coronary artery or rheumatic heart disease, left ventricular dysfunction, type of underlying arrhythmia, hypokalemia, hypomagnesemia, bradycardia, concomitant digoxin treatment, or level of QTc at baseline or while receiving drug. When cases of torsades de pointes were analyzed by individual drug, observed female prevalence was always greater than expected, representing a statistically significant difference (P < .05) for all agents except procainamide. CONCLUSIONS: These findings strongly suggest that women are more prone than men to develop torsades de pointes during administration of cardiovascular drugs that prolong cardiac repolarization. The pathophysiological basis for, and therapeutic implications of, this gender disparity should be further investigated. PMID- 8230645 TI - Users' guides to the medical literature. II. How to use an article about therapy or prevention. A. Are the results of the study valid? Evidence-Based Medicine Working Group. PMID- 8230646 TI - Silicone gel breast implants. Council on Scientific Affairs, American Medical Association. PMID- 8230647 TI - A call for higher standards for breast implants. PMID- 8230648 TI - The breast implant controversy. A clash of ethics and law. PMID- 8230649 TI - Depression and HIV. How does one affect the other? PMID- 8230650 TI - The AMA-MSS and AMSA: diversification, not division. PMID- 8230651 TI - Tuberculosis: the continuing story. PMID- 8230652 TI - Small, perfect circles. PMID- 8230653 TI - Parapneumonic effusions and thoracocentesis indications and complications. PMID- 8230654 TI - Factor XIII deficiency in Pakistan. AB - Patients with undiagnosed haemostatic defects seen at The Aga Khan Hospital and Fatimid Blood Transfusion Centre during the period of 7 years (1985-1992) were screened with routine tests including bleeding time (BT), whole blood clotting time (CT), platelet count, activated partial thromboplastin time (APTT), prothrombin time (PT) and 5 molar urea test. Nine patients had a positive 5 molar urea test indicating factor XIII deficiency. Rest of the screening tests were normal in these patients. High incidence of consanguinity was observed in affected families. Clinical features included excessive bleeding from umbilical stump, bruising, post-traumatic bleeding, epistaxis, melaena and intracerebral bleeding. All the patients were treated with fresh frozen plasma and cryoprecipitate. PMID- 8230655 TI - Psychiatric morbidity: prevalence, associated factors and significance. AB - A cross-sectional observational systematic study was carried out on ambulatory patients at a tertiary care hospital to determine the probable prevalence, associated factors and significance of psychiatric morbidity by using an Urdu translation of the hospital anxiety and depression (HAD) scale over a period of 6 days in a week. Results showed a prevalence of 38.4% which is slightly higher than what has been generally reported (30%). Two variables, i.e., female sex and being a housewife were significantly related with the outcome. An attempt has been made to identify the probable reasons for this and some suggestions laid down for further work. PMID- 8230656 TI - Changing trends in abdominal trauma in a teaching hospital of Lahore. AB - A comparison between patients of abdominal trauma admitted 10 years ago (1979 1981) and those admitted in the recent past (1989-1991) showed important variations in the frequency, aetiology, severity, morbidity and mortality. There was marked increase in frequency over this period. Ten years ago stab injuries were the most frequent cause of abdominal trauma, today firearm injuries rank first. The severity of injuries judged by involvement of number of organ systems and associated injuries has also intensified. Morbidity and mortality has almost doubled over this period. Today's emergency surgeon needs to be better trained and competent to deal with the severe injuries particularly of deep seated intra abdominal organs if mortality and prolonged morbidity is to be decreased. PMID- 8230657 TI - Pectoralis major myocutaneous flap for reconstruction of defects following resections in head and neck area. AB - Pectoralis major myocutaneous flap (PMMF) has become the standard for reconstruction of major defects in head and neck area. Eleven cases, operated over a three year period, in which PMMF was used for reconstruction have been reviewed retrospectively. Nine patients had oral squamous cell carcinoma, one had a basal cell carcinoma of the external ear and one had lost skin and soft tissue of neck following synergistic gangrene. Ten of the eleven flaps survived (success rate 91%). One of the three rib grafts used to reconstruct mandible got infected and had to be removed. Three patients developed wound infections and one had a temporary orocutaneous fistula which closed spontaneously. This brief experience confirms the reliability and efficiency of PMMF for head and neck reconstruction. PMID- 8230658 TI - Role of magnesium in the management of hypertension. AB - Twenty-two patients receiving long term diuretic treatment for arterial hypertension (19) and congestive heart failure (3) received magnesium chloride 10 mmol/day for four months. Both systolic and diastolic pressures decreased significantly, by a mean of 13 +/- 9 mmHg. No significant changes were recorded in serum or urinary electrolytes except for magnesium. PMID- 8230659 TI - Cerebral amoebiasis. PMID- 8230660 TI - The ring butterfly fragment. PMID- 8230661 TI - Acute intestinal obstruction due to hen's egg. PMID- 8230662 TI - Prenatal diagnosis. PMID- 8230663 TI - The value of intermittent growth monitoring in primary health care programmes. AB - The objective of growth monitoring is to detect early growth faltering before the child becomes severely malnourished. It has been found in many large projects in the private sector and in Government run Primary Health Care (PHC) programmes that monthly weighing is not feasible which makes it impossible to develop a functional outreach programme on a sustainable basis. This study is an attempt to propose an intermittent growth monitoring which is operationally feasible for large scale PHC programmes in the public sector. A historical prospective study was conducted to find out the correlation of weights of children at different ages with weights at subsequent months. Two hundred and ninety-two growth cards of children were selected from two squatter settlements of Karachi which are having a PHC programme through the Aga Khan University. At six months, about 71% of children were within the normal range, with increasing age from 10% to 39% of these children shifted to grade I Protein Energy Malnutrition (PEM). When weights of children for each month were correlated with weights at all other months up to 24 months, it was found that correlation coefficient at 6th and 9th month were significant (P value < 0.001). Probability of developing malnutrition at different weights and ages were also calculated. The results indicate that malnutrition starts appearing at 6 months and weights at 6 months and to a lesser extent at 9 months are better prognostic indices of future malnutrition. Intermittent weighing of children can help in early identification of "high risk" children who can then be managed and even be prevented from developing future malnutrition. PMID- 8230664 TI - Percutaneous radiofrequency retrogasserian rhizotomy for trigeminal neuralgia. PMID- 8230665 TI - Microvascular decompression for trigeminal neuralgia. AB - Eleven patients with intractable trigeminal neuralgia were treated by posterior fossa microvascular decompression. Complete pain relief was achieved in 7 patients (63.3%). Partial relief of symptoms was noted in 3 patients (27%). There was no operative mortality. Two patients developed transient lower cranial nerve palsies whereas one patient developed a CSF leak, requiring treatment by lumbar drain. Compared to ablative procedures, microvascular decompression is an effective and safe method of treatment for trigeminal neuralgia. PMID- 8230666 TI - Penile constrictive band injury. AB - Penile strangulation caused by a thread or human hair is an uncommon cause of urethral injury described in paediatric practice. Five children presented with this entity to the Aga Khan University Hospital, Karachi between August, 1991 and August, 1992. Three children had uneventful recoveries after removal of the constricting agent. Two patients developed urethrocutaneous fistulae and partial amputation of the penis because of late presentation. Early recognition and removal of the constricting agent is necessary to prevent serious complications. PMID- 8230667 TI - Diabetic ketoacidosis in a hospital based population in Pakistan. AB - Sixty-two consecutive episodes of diabetic ketoacidosis (DKA) were studied at Aga Khan University Hospital, Karachi. Forty-four (71%) were type I and 18 (29%) type II diabetics. Mean age was 28.1 years and mean duration of diabetes 4.1 years. Infections were the most common precipitating factor accounting for 28 episodes (45.2%). Twenty-two patients (35.5%) had hyperosmolality (serum osmolality > 320 mosmol/L). Mean serum Na+ was 131.7 mmol/L and K+ 4.6 mmol/L. Twenty-three (37.1%) were hyperkalemic at presentation with seven patients (11.3%) being comatosed and 35 (56.5%) alert. Mean random blood glucose (RBG) was 624 mg/dl, mean pN 7.09, osmolality 316 mosmol/L and the neurological status correlated statistically significantly with mean RBG, pH and osmolality. A leukemoid response was seen in 83.9% episodes. Mortality rate was 8.0% in patients with DKA managed in this hospital. PMID- 8230668 TI - Comparison of commercially available CLO test with the locally prepared test. PMID- 8230669 TI - Escobar syndrome. PMID- 8230670 TI - Malnutrition-related diabetes mellitus (MRDM)--an uncommon disorder in Pakistan. PMID- 8230671 TI - An animal model of coronary thrombosis and thrombolysis--comparisons of vascular damage and thrombus formation in the coronary and femoral arteries after balloon angioplasty. AB - The purpose of this study was to compare vascular damage and thrombus formation in the coronary and femoral arteries after balloon angioplasty, and to develop a physiological animal model of intracoronary occlusive thrombus using the balloon angioplasty technique. Angioplasty of the left anterior descending coronary arteries of 14 dogs was performed with an oversized balloon catheter at a high inflation pressure (150 PSI). This was followed angiographically (PTCA protocol). Dogs that showed arterial occlusion were divided into 2 groups. The dogs in 1 group were killed with an overdose of sodium pentobarbital, and those in the other group were infused with a tissue-type plasminogen activator (t-PA; 300,000 unit/kg). Angioplasty of the femoral and profunda femoris arteries (n = 5) was performed in 5 other dogs (PTA protocol). All of the animals were eventually sacrificed and tissue preparations were made from all 3 types of arteries. In the PTCA protocol, acute arterial occlusion was seen angiographically within 2 h in 10 of the 14 dogs. A histological study of the acutely occluded arteries (n = 5) showed thrombotic occlusion and severe arterial damage with medial tearing. T-PA was infused to 5 of the dogs with acute occlusion, and all showed reperfusion. A histological study of these animals showed severe arterial damage, but no macroscopic thrombus. In 4 dogs without acute occlusion, none of the 10 arteries examined were acutely occluded. In the PTA protocol, none of the 10 arteries were acutely occluded. A histological study showed fewer thrombi and less severe arterial damage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230672 TI - Effects of hydrogen peroxide on stimulatory guanine nucleotide-binding protein in rat heart. AB - This study was undertaken to examine the effects of hydrogen peroxide on stimulatory guanine nucleotide-binding protein (Gs), and coupling in the beta adrenergic receptor-Gs-adenylate cyclase system in rat heart, in vitro. Cardiac membranes were preincubated with various concentrations (0.1, 1, and 10 mM) of hydrogen peroxide at 30 degrees C for 5, 10, 30 and 60 min. Although the assay of beta-adrenergic receptors involving [3H]-dihydroalprenolol ([3H]-DHA) binding revealed that the maximal number of binding sites (Bmax) was not altered, the dissociation constant (Kd) for [3H]-DHA was increased in the presence of 1 mM and 10 mM hydrogen peroxide (control 0.68 +/- 0.16 nM, vs 1 mM H2O2 1.13 +/- 0.16, 10 mM H2O2 1.01 +/- 0.12). Conversely, no significant changes in Gs activities were observed in hydrogen peroxide-treated groups. Adenylate cyclase activity (stimulated by forskolin) was significantly reduced by 10 mM hydrogen peroxide after a 5 min preincubation period (control 277.1 +/- 19.2 pmol cAMP/mg protein/min, H2O2 230.3 +/- 14.9). The amounts of cyclic AMP produced by the stimulation of membranes with GTP, GTP+(l)-isoproterenol, guanylimidodiphosphate (Gpp(NH)p) or Gpp(NH)p+(l)-isoproterenol were significantly lower in 10 mM hydrogen peroxide-treated groups than those in controls (GTP: control 57.6 +/- 5.6 pmol cAMP/mg protein/min vs H2O2 46.4 +/- 6.9, GTP+(l)-isoproterenol: control 83.9 +/- 10.2 vs H2O2 67.7 +/- 10.3, Gpp(NH)p: control 77.5 +/- 8.8 vs H2O2 61.0 +/- 8.6, Gpp(NH)p+(l)-isoproterenol: control 105.0 +/- 13.1 vs H2O2 83.9 +/- 12.2, forskolin: control 223.2 +/- 13.8 vs H2O2 182.8 +/- 18.4).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230673 TI - Acute myocardial infarction due to coronary embolization from left atrial myxoma. AB - We encountered a 67-year-old woman with a left atrial myxoma which was discovered during echocardiographic examination and emergency coronary arteriography just after an onset of acute inferior myocardial infarction. Coronary arteriography disclosed an abrupt and total occlusion of the right coronary artery and an abnormally large and tortuous atrial circumflex branch feeding a left atrial mass. These findings were the most useful for diagnosis. Aorto-coronary bypass surgery and excision of the myxoma were performed simultaneously by emergency operation. The postoperative course was uneventful. Myocardial infarction in this patient is believed to have been caused by coronary embolization from the left atrial myxoma. PMID- 8230674 TI - Coronary stenosis and steal phenomenon in coronary-pulmonary fistula--assessment with stress thallium tomography after coronary angioplasty and fistulectomy. AB - We present a 46-year-old male with unstable angina and bilateral coronary-to pulmonary artery fistulae in whom reversible myocardial ischemia was detected by exercise-stress thallium-201 single-photon emission computed tomography (SPECT). Coronary angiography revealed a 99% stenosis at the proximal site of the left descending coronary artery and bilateral coronary-to-pulmonary artery fistulae with a saccular aneurysm. Percutaneous transluminal coronary angioplasty abolished chest pain and electrocardiographic changes. However, definitely improved, but still present, stress-induced perfusion abnormalities were demonstrated by an exercise-stress thallium-201 SPECT study. Myocardial ischemia was the only serious complication related to the coronary fistulae, and after they were surgically resected, the reversible perfusion abnormality was no longer observed. These findings suggest that coronary-to-pulmonary artery fistulae potentiated the myocardial ischemia in patient with coronary stenosis leading to unstable angina and prolonged the presence of coronary perfusion abnormality on stress thallium scans probably through a coronary steal phenomenon. PMID- 8230675 TI - Effects of vasodilators on pulmonary venous and mitral flow velocity patterns in patients with congestive heart failure. AB - To characterize abnormalities in pulmonary venous flow velocity patterns and to explore the changes in these patterns following short-term (3 to 14 days) administration of vasodilators, pulmonary venous and mitral flow velocity patterns were serially studied at congestive heart failure and after vasodilator administration in 18 patients. Peak mitral early diastolic filling velocity (E) and the ratio of E to peak filling velocity at atrial contraction (E/A) consistently decreased after vasodilator administration by 30 +/- 4 cm/s and 0.74 +/- 0.13 (mean +/- SD), respectively. Peak pulmonary venous diastolic forward flow velocity also decreased by 29 +/- 4 cm/s. However, changes in peak pulmonary venous systolic forward flow velocity (S, cm/s) did not correlate with changes in E, the E/A ratio, or D (peak pulmonary venous diastolic forward flow velocity). Thus, when patients were divided into two groups on the basis of changes in S, indices of left ventricular systolic performance, such as end-systolic dimension and fractional shortening, improved more in the group which showed an increase in S after vasodilator administration than in the group which showed a decrease in S (-7 +/- 6 vs -1 +/- 4 mm, p < 0.05; 8 +/- 6 vs -1 +/- 4%, p < 0.05). Although the mitral flow velocity pattern changed uniformly with vasodilator administration in patients with mild to moderate congestive heart failure, the changes in pulmonary venous flow velocity patterns were not uniform among patients. Pulmonary venous flow velocity patterns appear to reflect changes in left ventricular systolic performance in addition to those in left ventricular diastolic performance. PMID- 8230676 TI - Enhanced elevation of blood pressure during cigarette smoking in the elderly. AB - Acute changes in blood pressure and heart rate during cigarette smoking were investigated in 10 elderly subjects aged 75.6 +/- 4.8 (mean +/- SD) years and in 10 non-elderly subjects aged 35.6 +/- 10.5 years. All subjects were healthy normotensive men with no cardiovascular disease. While each subject smoked two cigarettes containing 5.4 mg of nicotine, systolic and diastolic blood pressures increased significantly more in the elderly subjects (18.2 +/- 6.0/9.0 +/- 5.0 mmHg) than in the non-elderly subjects (4.8 +/- 4.7/3.9 +/- 4.2 mmHg, p < 0.05). However, the increases of heart rate in the two groups were similar (elderly: 10.5 +/- 4.1 beats/min versus non-elderly: 12.1 +/- 6.4 beats/min, ns). These results indicate that there is an age-related difference in blood pressure elevation during cigarette smoking in healthy subjects. PMID- 8230677 TI - Long-term results of catheter ablation for idiopathic ventricular tachycardia originated from the right ventricular outflow. AB - We performed catheter ablation in 10 consecutive patients with idiopathic monomorphic right ventricular tachycardia and studied the long-term outcome. All ventricular tachycardias had a left bundle branch block configuration with an inferior axis, and originated from right ventricular outflow. Antiarrhythmic drugs (3-6 drugs) had been ineffective in controlling ventricular tachycardia. The 2 patients who underwent direct-current ablation (2 shocks of 150 J) had no recurrence of ventricular tachycardia and did not require antiarrhythmic drugs during a follow-up of 56 and 51 months, respectively. Of the 8 patients who underwent radiofrequency ablation (30-40 watt, 20-40 sec, 2-15 application, using Inter Nova RA 50, 13.56 MHz), 1 patient had no recurrence of ventricular tachycardias and did not require antiarrhythmic drugs, 4 patients had no recurrence of ventricular tachycardias but did require anti-arrhythmic drugs, and 3 patients experienced recurrence of non-sustained ventricular tachycardia despite the use of antiarrhythmic drugs during a follow-up of 15-40 months. There were no complications except for cardiac perforation which occurred immediately after direct-current ablation in 1 patient. In conclusion, long-term success in preventing ventricular tachycardia was achievable with direct-current ablation, but this success was associated with serious risks, such as cardiac perforation. Radiofrequency ablation was safer than direct-current ablation, but had a lower long-term success rate. PMID- 8230678 TI - Response of post-ischemic myocardium to sympathetic stimulation--relation to local norepinephrine release. AB - Myocardial ischemia interrupts neurotransmission and causes the depression of norepinephrine release. However, the effects of sympathetic nerve stimulation on neurotransmission and norepinephrine release in post-ischemic myocardium are not well defined. We measured regional myocardial length and norepinephrine (NE) release during sympathetic nerve stimulation in anesthetized dogs. Dogs were divided into 2 groups: Group 1 (n = 14); sympathetic nerve stimulation, Group 2 (n = 9); pre-treatment with alpha-blockade yohimbine hydrochloride (0.2 mg/kg) followed by sympathetic nerve stimulation. The left anterior descending artery was occluded for 15 min. Sympathetic nerve stimulation was performed before coronary occlusion and after reperfusion. In group 1, the decrease in systolic shortening in the ischemic region persisted for more than 60 min. Although sympathetic nerve stimulation caused an increase in systolic shortening, it was lower than the pre-ischemic value. NE release from the post-ischemic myocardium remained decreased for 60 min. The decrease in the post-ischemic myocardial response to sympathetic nerve stimulation was associated with diminished NE release. When the cardiac sympathetic nerve was denervated with an epicardial phenol application, NE release decreased even further. In group 2, NE release did not decrease following reperfusion. These results suggest that sympathetic nerve conduction is not completely impaired in post-ischemic myocardium and that pre synaptic alpha -2 receptors might play an important role in diminished NE release. PMID- 8230679 TI - Antithrombotic effects of BMY21190, an inhibitor of cAMP phosphodiesterase, in a canine model of coronary artery thrombosis. AB - BMY21190, an inhibitor of cyclic AMP phosphodiesterase, has a coronary vasodilating effect. BMY21190 was evaluated for its ability to modify the development of experimental thrombosis resulting from anodal current injury (100 microA for 6 h) of the intimal surface of the left circumflex coronary artery (LCX) in anesthetized dogs. Two groups of dogs were studied. One group received BMY21190 (1 mg/kg) and the other group received an equal volume of vehicle infused into the left jugular vein. After a 30 min administration of BMY21190, heart rate and mean coronary blood flow were increased significantly and mean arterial pressure was decreased. However, the myocardial tension of the left anterior descending coronary artery (LAD) and LCX areas did not increase significantly after BMY21190 infusion. During LCX stimulation, the first LCX occlusion and hyperemic reaction of the control group both occurred significantly earlier than those of the BMY21190 group. BMY21190 treatment reduced the development of the LCX thrombus mass, as compared to that in the controls. In ex vivo studies, platelet aggregation in response to arachidonic acid, ADP or collagen was inhibited by BMY21190. These results suggest that BMY21190 possesses anti-thrombotic and coronary vasodilating effects which may be mediated through the inhibition of cyclic AMP phosphodiesterase. PMID- 8230680 TI - Frequency analysis of QRS complex with bundle branch block in patients with and without sustained ventricular tachycardia. AB - To distinguish patients with bundle branch block (BBB) and sustained ventricular tachycardia (s-VT) from patients with BBB but without s-VT, a frequency analysis of the QRS complex was performed in 71 patients. Frequency analysis of the QRS complex of patients with left bundle branch block (LBBB) showed that patients with s-VT had significantly larger areas and area ratios between 50 and 100 Hz in the X lead than patients without s-VT (area: -0.905 +/- 0.231 vs -1.195 +/- 0.286. area ratio: -0.783 +/- 0.230 vs -1.125 +/- 0.310; P < 0.05). The area and area ratios from 100 to 200 Hz in the Z lead were also larger in patients with s VT. The highest predictive accuracy using the area ratio from 50 to 100 Hz in the X lead was 86%, with a sensitivity and specificity of 83% and 88%, respectively. In cases with LBBB, time domain analysis showed no significant difference between patients with s-VT and those without s-VT. Frequency analysis of the QRS complex may be useful for distinguishing LBBB patients with s-VT from those without s-VT. PMID- 8230681 TI - Detection of coronary artery stenosis by dipyridamole radionuclide ventriculography. AB - We assessed the usefulness of dipyridamole radionuclide ventriculography for detecting significant coronary artery stenosis in 89 patients who were undergoing cardiac catheterization. Radionuclide ventriculography was performed before and after the infusion of dipyridamole (0.56 mg/kg). The end-diastolic regions of interest of the left ventricle were divided into 5 sectors for calculation of the regional ejection fractions. Results were considered to be positive when the regional ejection fraction decreased by more than 5% after the infusion of dipyridamole. The presence of significant coronary artery stenosis (> 75%) was demonstrated by arteriography in 49 patients and was absent in 40 patients. A decrease in the regional ejection fraction greater than 5% was observed in 41 (84%) of the 49 patients with significant coronary artery stenosis and in 2 of the 40 without significant coronary stenosis. The sensitivity and specificity of this method for detecting significant coronary artery stenosis were 84% and 95%, respectively. We conclude that a decrease in the radionuclide-determined regional ejection fraction after the infusion of dipyridamole reflects left ventricular dysfunction and is a sensitive and specific indicator of significant coronary artery stenosis. PMID- 8230682 TI - Comparative effects of early and late reperfusion of the infarct-related coronary artery and collateral circulation which develops after infarction on left ventricular size and function. AB - This study assesses the effects of early and late reperfusion of the infarct related coronary artery and collateral circulation developed after infarction on left ventricular morphology and function in 22 patients with a first acute anterior myocardial infarction and without long-standing preinfarction angina. The patients were categorized into 4 groups: group A-5 patients with successful reperfusion within 6 h after the onset of infarction: group B-6 patients with late patency of the infarct-related coronary artery; group C-6 patients without recanalization of the infarct-related coronary artery who had good collateral circulation (collateral index (CI) = 2 or 3); and group D-5 patients without recanalization who had either poor or no collateral circulation (CI = 0 or 1). The left ventricular size and function were evaluated by means of contrast left ventriculography during the chronic stage of infarction. The left ventricular global and regional functions were preserved better (p < 0.05) in group A than in the other three groups. Although there was no difference in regional wall motion of the infarct area among groups B, C and D, the ejection fraction was greater (p < 0.05) and the percentage perimetric circumference was smaller (p < 0.05) in groups B and C than in group D. Among the 4 groups, the left ventricular end diastolic volume index tended to be greater in group D. Thus, these results suggest that blood supply to the infarct area is essential for the preservation of left ventricular size and function regardless of the timing and route. PMID- 8230683 TI - Estimation of infarct size using serum troponin T concentration in patients with acute myocardial infarction. AB - To estimate the size of myocardial infarction, serum troponin T concentration was measured in 34 patients with acute myocardial infarction. Left ventriculography, 2-dimensional echocardiography and resting 201thallium myocardial single photon emission computed tomography (SPECT) were performed about 4 weeks after the onset of myocardial infarction and used for correlation with the late serum troponin T peak concentration which occurred on the 3rd to 5th day after onset. Both left ventricular ejection fraction (LVEF) obtained from left ventriculography and wall motion index (WMI) obtained from 2-dimensional echocardiography were inversely related to late troponin T peak value (LVEF: r = 0.68, p < 0.001, WMI: r = 0.70, p < 0.001). Extent score (ES) and severity score (SS), which were estimated from the initial resting 201thallium SPECT image, showed excellent linear correlations with late troponin T peak concentrations (ES: r = 0.77, p < 0.001, SS: r = 0.66, p < 0.001). This correlation was present both in patients with an early troponin T peak on day 1 (group A-16 patients) and in those without an early peak (group B 10 patients). Thus, late troponin T peak concentration can be used to predict infarct size regardless of the kinetics of its appearance in serum. PMID- 8230684 TI - Surgical treatment of active infective endocarditis--early and late results of active native and prosthetic valve endocarditis. AB - The purpose of this study was to determine the clinical predictors of active infective endocarditis in 45 cases we treated between January 1971 and August 1991 (30 native valve endocarditis (NVE) and 15 prosthetic valve endocarditis (PVE). The indication of surgery in 45 patients was progressive congestive heart failure (CHF), septicemia and systemic embolization. The aortic valve was involved in 24 (53%) of 45 patients (13 of 30 NVE and 11 of 15 PVE) and there was significantly higher early mortality in aortic PVE (36%) than in aortic NVE (8%). The 9 patients with severe cardiac failure (NYHA Class V) before surgery were associated with a significantly higher incidence of early mortality (5/9 = 56%) than those in Class III (2/14 = 14%) and Class IV (3/18 = 17%). We concluded that aortic valve infection is more prevalent than mitral valve infection and is more often associated with staphylococcus infection, including abscess formation. Early surgical intervention should be performed despite the risk of cardiac failure and extensive infection. PMID- 8230685 TI - Dual-tracer autoradiography with thallium-201 and iodine-125 MIBG in BIO 14.6 cardiomyopathic Syrian hamsters. AB - Dual-tracer imaging of the heart with 125I-metaiodobenzylguanicline (MIBG) and 201Tl can simultaneously demonstrate the distribution of sympathetic nerve endings and the underlying myocardial perfusion. A quantitative dual-tracer autoradiographic study with 201Tl and 125I-MIBG was performed to investigate changes in the distribution of cardiac sympathetic innervation with the progression of cardiomyopathy in BIO 14.6 hamsters. The distribution of 201Tl was uniform in control hamsters and BIO 14.6 hamsters at all stages of cardiomyopathy. In contrast, a reduction in MIBG accumulation occurred in the endocardial region of the left ventricular free wall and the left ventricular aspect of the interventricular septum in BIO 14.6 hamsters at 3 and 8 months of age. Thus, there was an uncoupling of the left ventricular distribution of 201Tl and 125I-MIBG in BIO 14.6 hamsters. In addition, interstitial fibrosis was increased in the interventricular septum, the subendocardial region of the left ventricular free wall, and the right ventricular wall, which were the sites of reduced MIBG accumulation. This study shows that dual myocardial imaging with MIBG and 201Tl may be useful for investigating patients with cardiomyopathy. PMID- 8230686 TI - Effects of renal denervation on pressure-natriuresis in spontaneously hypertensive rats. AB - To investigate the role of renal sympathetic nerve activity (RSNA) under developing and established hypertension, renal function was studied in chronically renal-denervated and sham-operated male spontaneously hypertensive rats (SHR) and control Wistar Kyoto rats (WKY) at 8 (early hypertensive) and 22 (established hypertensive) weeks of age. To further characterize the renal pressure-natriuresis-diuresis relationship in SHR, renal perfusion pressure (RPP) was reduced by aortic constriction to the level seen in age-matched WKY and the same studies were repeated. After denervation, urinary sodium excretion (UNaV), fractional excretion of sodium (FENa) and urine flow (UF) were increased in 8 week-old SHR (p < 0.01). With the exceptions of UNaV and FENa in denervated 8 week-old SHR, renal cortical blood flow, glomerular filtration rate, UF, UNaV and FENa decreased with the reduction of RPP in all of the SHR groups. These results suggest that RSNA significantly influences renal sodium and fluid handling, thus contributing to the shifting of the arterial pressure-renal sodium excretion curve to the right along the pressure axis and/or to an increase in the steepness of the relationship in 8-week-old SHR. There appeared to be a marked difference in renal sodium handling between 8- and 22-week-old SHR. PMID- 8230687 TI - Successful treatment of life-threatening ventricular tachycardia with high-dose propranolol under extracorporeal life support and intraaortic balloon pumping. AB - The use of propranolol is generally contraindicated in patients with congestive heart failure. However, we successfully used a continuous high-dose infusion of propranolol, in combination with the use of extracorporeal life support (ECLS) and intra-aortic balloon pumping (IABP), to treat refractory life-threatening ventricular tachycardia in a patient with postoperative congestive heart failure. The early combined use of ECLS and IABP before irreversible myocardial damage contributed to the successful outcome. PMID- 8230688 TI - [An application of statistics to closed circuit anesthesia practice]. PMID- 8230689 TI - [Effects of sevoflurane anesthesia on serotonin metabolism in rat brain]. AB - To elucidate the mechanism of general anesthesia, effects of sevoflurane anesthesia on serotonin metabolism in rat brain were studied. Three percent sevoflurane was administered for twenty minutes to Wistar male rats weighing 230 270g under spontaneous respiration. The rats were sacrificed by decapitation and the brains were rapidly removed. They were dissected into nine discrete regions, locus coeruleus, pons plus medulla oblongata, hypothalamus, thalamus, basal ganglia, midbrain, hippocampus, amygdala and cerebral cortex. The contents of serotonin (5-HT) and one of its major metabolites, 5-hydroxyindole-3-acetic acid (5-HIAA) were measured by high performance liquid chromatography with the dual cell coulometric detector before anesthesia, 20 minutes after the start of anesthesia and at the recovery from anesthesia. Significant increases in 5-HT levels were observed in the pons, hypothalamus, midbrain, amygdala and cerebral cortex by sevoflurane anesthesia as compared with the control group. 5-HIAA levels decreased significantly in the thalamus by sevoflurane anesthesia, while an appreciable increase in 5-HIAA levels was observed in the basal ganglia at the recovery from anesthesia. It is concluded that 5-HT metabolism is significantly suppressed in the pons, hypothalamus, thalamus, midbrain, amygdala and cerebral cortex during sevoflurane anesthesia and this change in 5-HT metabolism may be associated with a mechanism of sevoflurane anesthesia. PMID- 8230690 TI - [The effects of sevoflurane anesthesia with hypothermia on ischemic brain cellular respiration]. AB - We examined the effects of sevoflurane anesthesia with hypothermia on ischemic brain cellular respiration using Wistar strain rats. Ischemia was induced by decapitation. L/P in brain increased during no anesthesia-normothermia-ischemia (1), then gradually, but not fully recovered during 2.5% sevoflurane anesthesia normothermia-ischemia (2) and during 2.5% sevoflurane anesthesia-hypothermia ischemia (3). Energy charge decreased during (1) and slight recovery was observed during (3). Respiratory control ratio in brain mitochondria decreased during (1) and recovery during (2) and (3) was slight. No recovery was observed in ADP/O. These findings indicate that during sevoflurane anesthesia with hypothermia, ischemic brain energy metabolism has an effect not on the mitochondrial energy producing processes but on the consuming system. PMID- 8230691 TI - [Intranasal administration of nitroglycerin solution and nitroglycerin spray during general anesthesia]. AB - Nitroglycerin spray is a newly developed nitroglycerin (TNG) form, which delivers 0.3 mg of TNG with each spray emission. We compared the efficacy of intranasal administration of TNG spray and that of TNG solution during general anesthesia. Nineteen patients were randomly assigned to receive either 0.3 mg TNG solution (Solution group) or 0.3 mg TNG spray (Spray group) intranasally. Peak elevation of plasma TNG occurred 2 min after drug administration in both groups, and the plasma level decreased to an ineffective level after 10 min. The plasma TNG of the spray group was significantly higher than that of the solution group for 3 min. Two min after drug administration, systolic blood pressure decreased significantly in both groups, and this level persisted for 10 min. Diastolic blood pressure also decreased in the spray group, but not in the solution group. Heart rate increased significantly in both groups 2 min after drug administration. Oxygen index (PaO2/FIO2) decreased significantly in both groups, and the level persisted 10 min after administration. Although no significant differences were found between two nitroglycerin forms in any of these hemodynamic and respiratory parameters, nitroglycerin spray may have clinical advantages because of reliability in its effect, ready availability, ease of application and high stability. PMID- 8230692 TI - [Is the baroreflex heart response to prostaglandin E1 less than that to nitroglycerin?]. AB - The purpose of this study is to investigate and compare the reflex heart rate response to bolus injections of prostaglandin E1 (PGE1) and to nitroglycerin (NTG). One hundred and fifty seven patients under isoflurane nitrous oxide in oxygen anesthesia were studied prior to elective surgery. They were divided into following two groups: NTG group; n = 101 (male 54, female 47), 49.8 +/- 19.1 years of age, and PGE1 group; n = 56 (male 31, female 25), 54.7 +/- 15.3 years of age. The bolus injections of NTG and PGE1 produced a significant increase in heart rate as an indicator of arterial baroreflex response. However, there was no statistical difference between the two groups. Negative linear relationship between age and magnitude of reflex heart rate response to reduction of blood pressure (delta HR/delta SBP) was present in NTG group (r = -0.57, P < 0.01) and PGE1 group (r = -0.50, P < 0.01). These results suggest that the degree of the reflex heart rate response to bolus injection of NTG and PGE1 is almost identical in patients under isoflurane anesthesia. PMID- 8230693 TI - [Effects of nicardipine on hemodynamics and skin blood flow--comparison between N2O-sevoflurane and N2O-isoflurane anesthesia]. AB - Dividing the surgical patients into GOS (N2O-oxygen-sevoflurane anesthesia) group and GOI (N2O-oxygen-isoflurane anesthesia) group, we studied the effects of nicardipine on hemodynamics and skin blood flow under each anesthetic method. Control measurements were taken when the hemodynamic parameters were stable during surgery and the incremental doses of nicardipine (0.5 mg) were administered up to a total of 2.0 mg. The changes in hemodynamic parameters and skin blood flow were measured at the nicardipine dosage of 0.5 mg (S1), 1.0 mg (S2), 1.5 mg (S3), and 2.0 mg (S4). Heart rate tended to increase in both groups. Blood pressure, on the other hand, decreased at S1 and S2, but showed little change at S3 and S4 in both groups. Skin blood flow tended to increase in the GOS group, but in the GOI group little change in skin blood flow was noted at S1-S3. Skin blood flow tended to decrease at S4 in the GOI group. PMID- 8230694 TI - [Effect of ketamine on hepatic circulation and oxygen metabolism during hypoxia in dog]. AB - The present study was designed to determine whether ketamine could maintain hepatic circulation and oxygen metabolism even in the presence of serious hepatic hypoxia. Hepatic oxygen delivery as well as consumption and the hepatic energy charge were assessed in twenty mongrel dogs receiving 30 mg.kg-1.h-1 of ketamine or thiamylal intravenously while inhaling graded hypoxic gas mixtures (FIO2 0.21 to 0.08). Hepatic blood flow was measured using electromagnetic flowmetry: hepatic oxygen delivery and consumption were calculated from hepatic blood flow and oxygen content in hepatic arterial, portal venous and hepatic venous blood. The hepatic energy charge was assessed by measuring arterial ketone body ratio (AKBR). In the ketamine group, hepatic arterial blood flow was well maintained but portal venous blood flow decreased significantly at FIO2 0.21 and 0.12 compared with thiamylal group. At FIO2 0.08, hepatic oxygen delivery, consumption and AKBR decreased significantly in the ketamine group mainly due to the decrease in oxygen delivery from portal vein, with concomitant increase in catecholamine level. These results suggest that ketamine could not suppress the exaggerated reaction to hypoxemia, and it could not maintain hepatic circulation, hepatic oxygen metabolism or hepatic mitochondrial redox state in better condition during serious hypoxemia compared with thiobarbiturate. PMID- 8230695 TI - [The effects of spinal anesthesia on liver blood flow measured by the ICG clearance meter]. AB - ICG (indocyanine green) clearance meter is a new equipment that can measure the plasma disappearance of ICG through a photopiece attached to the tip of the finger. In this study we evaluated the effects of spinal anesthesia on liver blood flow using this method in 12 adult patients who were scheduled for elective surgery under spinal anesthesia. ICG clearance was read from an ICG clearance meter RK-1000 (Sumitomo Electric) following intravenous administration of ICG (0.5 mg.kg-1). The data are shown as the plasma disappearance rate (K) and the 15 minute retention rate (R15). The measurements were performed prior to spinal anesthesia and when the sensory level of anesthesia reached Th5. Spinal anesthesia was performed with tetracaine (12-15 mg). Heart rate, arterial blood pressure, and cardiac output (measured by NCCOM) were recorded simultaneously. Mean arterial pressure, heart rate and ICG clearance decreased significantly but cardiac index was unchanged. Our results suggest that high spinal anesthesia causes circulatory disturbance of the liver, and that this is probably followed by a drop in arterial blood pressure. PMID- 8230696 TI - [Studies of emergency cardiopulmonary bypass (ECPB) for cardiopulmonary-cerebral resuscitation--(3). Neurological outcome and pathological changes of the brain]. AB - Neurological outcome and pathological changes of the brain were studied in 5 female mongrel dogs, which were subjected to normothermic ventricular fibrillation (Vf) cardiac arrest of 15 min and resuscitated by using cardiopulmonary bypass through the femoral artery and veins (F-F bypass). Spontaneous circulation was restored by one or two defibrillating countershocks in all 5 dogs 5.2 +/- 1.1 (mean +/- SD) min after initiation of the F-F bypass. The F-F partial bypass was continued for 164 +/- 30 min under mild hypothermia. After weaning from the bypass, intensive care including controlled ventilation was carried out for the subsequent 6 to 36 h. Intermittent slow waves appeared on the electroencephalogram 62.8 +/- 11.6 min after initiation of the F-F bypass resuscitation and continuous waves at 145.6 +/- 27.5 min. Soon after extubation, the animals barked, moved the forelegs and could drink water. Neurological deficit scores (normal: 0, brain death: 500) improved to become below 100 except in 1 dog. However, macroscopic examination of the brain in 2 dogs with prominent recovery disclosed atrophy of the central gyrus and microscopic examination revealed typical ischemic injuries of the vulnerable neurons at the cerebellum, hippocampus and cerebral cortex in the frontal lobe. PMID- 8230697 TI - [The alteration of right ventricular performance in patients with pneumonectomy and pulmonary lobectomy]. AB - Elevation of right ventricular afterload associated with the decrease of pulmonary vascular bed is a serious problem immediately after pneumonectomy. We studied the alteration of right ventricular performance and hemodynamics by using an RVEF thermodilution catheter, and compared them among 6 patients after pneumonectomy and 8 patients after pulmonary lobectomy. Results demonstrate that 1) PVRI increased significantly immediately after the pneumonectomy, and recovered to the preoperative level on the 1st postoperative day. On the other hand, in the lobectomy patients, PVRI showed no significant change throughout the operation, because the decrease of pulmonary vascular bed after pneumonectomy was larger than that after pulmonary lobectomy. 2) In the pneumonectomy group, CI and RVEF were preserved and right ventricular works (RCWI and RSWI) increased in spite of an increase of right ventricular afterload after the ligation of the pulmonary artery. We considered that immediately after pneumonectomy the increase of right ventricular afterload associated with decreased pulmonary vascular bed was compensated by increasing right ventricular work. 3) On the 1st postoperative day, CI and RVEF did not change, but mRAP decreased significantly, and PVRI and right ventricular works recovered to the preoperative level in the pneumonectomy patients. From the above results, we considered that after the pneumonectomy, increased right ventricular afterload was compensated by the decreased circulating blood volume. PMID- 8230698 TI - [Effects of epidural analgesia combined with general anesthesia on hemodynamics during neck surgery]. AB - The aim of the present study was to investigate the effect of epidural analgesia combined with general anesthesia on hemodynamics. Thirty patients undergoing surgery for the treatment of cancer of the neck were studied. The patients were divided into two groups of those who received epidural analgesia combined with general anesthesia group (Group 1) and those with general anesthesia alone (Group 2). Blood pressure was not different between the groups. But heart rate and rate pressure products in Group 1 were significantly lower than those of Group 2. CVP in Group 1 increased significantly to 10.1 +/- 2.9 mmHg during surgery from 6.8 +/- 1.8 mmHg at the beginning of the surgery. There was no difference in intraoperative blood loss and the amount of fluid infused between the two groups. These results suggest that epidural anesthesia combined with general anesthesia is effective to stabilize hemodynamics during cervical surgery, but we have to be careful about using local anesthetics during long cervical procedures, because it increases CVP which might result from the depression of cardiac function. PMID- 8230699 TI - [Influence of epidural and spinal block on VO2 and VCO2 measured by the indirect calorimetry]. AB - We evaluated the influence of epidural and spinal anesthesia on VO2 and VCO2 measured by the indirect calorimetry. According to the spreading of anesthetic level, VO2, VCO2 and EE increased in the epidural group (EP group), but they decreased in spinal group (SP group). RQ was unchanged in both groups. Rate pressure product (RPP), in proportion to the MVO2 and the REE, did not correlate with the EE. The VO2, VCO2 and EE in EP group increased probably because; (1) epidural anesthesia does not affect the VO2 change, as it does not markedly suppress respiration; (2) cardiac stroke work is retained during epidural anesthesia, as it does markedly suppress circulation. In conclusion, epidural anesthesia exerts less influence on the respiratory and circulatory systems of patients compared with spinal anesthesia. PMID- 8230700 TI - [Postoperative pain relief by continuous epidural infusion: a comparison of three solutions]. AB - The effect of three solutions on postoperative pain relief by continuous epidural infusion was studied. Seventy-five patients after upper or lower abdominal surgeries were assigned to one of three groups and the postoperative pain relief was evaluated for 48 hours. Group I: bolus injection of buprenorphine (Bn) 0.1 mg + saline (S) 8 ml and continuous infusion (2 ml.h-1) of Bn 0.8 mg + S 92 ml; Group II: bolus injection of Bn 0.1 mg + 0.5% bupivacaine (Bc) 4 ml + S 4 ml and continuous infusion of Bn 0.8 mg + S 92 ml; Group III: bolus injection of Bn 0.1 mg + 0.5% Bc 4 ml + S 4 ml and continuous infusion of Bn 0.8 mg + 0.5% Bc 40 ml + S 60 ml. The combination of buprenorphine with low-dose bupivacaine (Group III) offered the most effective postoperative analgesia in three groups without increasing the frequency of adverse effects induced by epidural administration of local anesthetics. However, even in Group III, the percentage of patients complaining of pain at bed rest was still high during early postoperative period (56% at 6 hours postoperatively). The results suggest that further consideration is necessary on agents selection and dosage adjustment for the postoperative epidural analgesia. PMID- 8230701 TI - [Changes in human evoked spinal potentials (ESP) by epidural lidocaine]. AB - Controversies exist on the sites of action of epidural anesthesia. Leading opinion says that it works on spinal nerve root. We examined ESP and tactile sensations in 4 patients during epidural anesthesia with lidocaine to determine the effects of the anesthesia on spinal cord. Prolongation of latency and decrease in amplitude of ESP appeared 10 min after injection of 1.5% lidocaine 4 ml, each parameter reaching maximum value of 115% and 60% of the control value 30 min afterwards, respectively. Then they started to recover slowly, reaching normal values 150 min later. Changes in ESP and tactile sensation were closely related. Decreases in amplitude to 90%, 80%, 60% of the control values were observed for hypesthesia, analgesia, and anesthesia, respectively. We conclude that the spinal cord also is an important site of action of epidural anesthesia. PMID- 8230702 TI - [Examination of the protein binding of lidocaine by gel filtration method]. AB - The protein binding of lidocaine has been studied using gel filtration method. Thermodynamic calculation has been done on results of the experiment. We have found that the primary bond type of lidocaine with serum albumin is hydrophobic interaction, because entropy (delta S) is constantly positive (> 0) under these experimental conditions. The change of the binding ratio between lidocaine and serum albumin with changing pH results from the conformational change of the serum albumin, but the protein binding of lidocaine is also an exothermic reaction. The following conclusions have been obtained. (1) The binding of serum albumin with lidocaine is nonspecific and mainly the result of hydrophobic interaction. (2) At temperature 35 degrees C and pH 7.5, binding constant (K) and binding site (np) of bovine serum albumin with lidocaine are K = 5.9 x 10(3) M-1 and np = 103.5 x 10(-6) M, respectively. (3) Increasing binding ratio with increase in pH is the result of the increase in binding site. This involves the conformational change of albumin, especially that of N-B transition. (4) The binding of serum albumin with lidocaine is an exothermic reaction. Therefore, the binding of lidocaine with serum albumin increases as the temperature decreases. PMID- 8230703 TI - [Spinorphin, a new inhibitor of enkephalin-degrading enzymes derived from the bovine spinal cord]. AB - We have isolated a potent inhibitor of enkephalin-degrading enzymes from the bovine spinal cord, and determined its amino-acid sequence and inhibitory activity against enkephalin-degrading enzymes. This new substance, isolated and identified from the bovine spinal cord, is composed of a heptapeptide (Leu-Val Val-Tyr-Pro-Trp-Thr). The inhibitory activity (IC50) of this new substance against enkephalin-degrading enzymes, purified from monkey brain, are 3.3 micrograms.ml-1 against aminopeptidase, 1.4 microgram.ml-1 against dipeptidyl aminopeptidase, 2.4 micrograms.ml-1 against angiotensin converting enzyme, and 10 micrograms.ml-1 against enkephalinase. This new substance showed no inhibitory activity against enkephalin-degrading enzymes purified from kidney, blood, etc. According to the above results, this substance is thought to be a new neuromodulator derived from the spinal cord. Because it was derived from the spinal cord, we have named it "Spinorphin". The discovery in the bovine spinal cord of endogenous heptapeptide, Spinorphin, with inhibitory activity on enkephalin-degrading enzymes raises a number of pertinent questions which cannot be adequately dealt with in this study. It will now be possible, however, to test the very hypothesis that this new peptide act as neurotransmitters or neuromodulators at synaptic junctions. PMID- 8230704 TI - [Open lung biopsy in an 11 year old boy with %FVC 11%]. AB - Anesthesia was administered to an 11 year old boy with %FVC 11%. The patient had dyspnea and severe constrictive pulmonary disorder with an almost normal chest radiograph. Open lung biopsy was performed and the patient was weaned in the first postoperative morning without any problem. Pulmonary fibrosis was caused by anti-cancer drugs given in his infancy. He died of cardiorespiratory failure 14 months postoperatively. PMID- 8230705 TI - [Anesthetic management for cesarean section of a patient with transient diabetes insipidus and acute severe liver dysfunction]. AB - A 26-year old woman presented with acute hepato-renal dysfunction, coagulation abnormalities and diabetes insipidus associated with hypernatremia in the latter term of pregnancy (39 weeks). Such transient diabetes insipidus during pregnancy as in this case has been reported to be resistant to AVP, but to respond to DDAVP. Because of fetal compromise, an urgent cesarean section was performed. Spinal anesthesia was chosen because of the possible deleterious effects of general anesthesia on liver function. After delivery of twin babies, her symptoms recovered gradually. In conclusion, diabetes insipidus during pregnancy as in this case is transient and disappear after delivery. However, multiple organ dysfunction may become worse and cause fetal death, unless surgical procedure with appropriate anesthetic management is performed. PMID- 8230706 TI - [Two cases of tension pneumothorax caused by tracheal suction during the operation of pectus excavatum]. AB - Rehbein's method is one of the surgical procedures for pediatric pectus excavatum, which constructs thorax by lifting and fixating the sternum using metal strut inserted into bone marrow of bilateral ribs. We experienced two pediatric cases of tension pneumothorax caused by tracheal suctioning during the operation by this method. The etiology of tension pneumothorax was thought to be as follows. A small pleural hole was made during separation of ribs from surrounding tissues and a large amount of air entered into the pleural space and check valve mechanism worked at the pleural hole when the lung was collapsed on tracheal suctioning. In anesthetic management of pectus excavatum by Rehbein's method, we should always be cautious of the possible occurrence of tension pneumothorax. PMID- 8230707 TI - [Anesthetic management of a patient with Bernard-Soulier syndrome]. AB - Bernard-Soulier syndrome (BSS) is a hereditary hemorrhagic disease characterized by prolonged bleeding time due to abnormal platelet aggregation and giant platelets. Transfusion of platelet-rich plasma is the only treatment available for the hemorrhagic episodes in patients with this disease. A 20-year-old female with BBS was scheduled for sagittal osteotomy of the mandibular rami under general anesthesia. Anesthesia was induced with fentanyl and diazepam, and was maintained with nitrous oxide, fentanyl, and 0.5% enflurane. No exacerbation of the bleeding tendency was observed during or after the surgery. We consider that the use of halothane should be avoided in patients with BBS because it may inhibit the aggregation of platelets and prolong the bleeding time. PMID- 8230708 TI - [Onset of RSD and causalgia]. AB - This study examined the time of onset of RSD and causalgia after injury. Many forms of treatment were tried to relieve the complex and debilitating symptoms in 36 subjects. Burning pain occurred in less than a month in 77.8% of patients. PMID- 8230709 TI - [Anesthetic management of a patient undergoing hemodialysis for coronary artery bypass surgery without extracorporeal circulation]. AB - A 58-year-old female with renal failure underwent anastomosis of the left internal mammary artery to the left anterior descending branch of coronary artery without extracorporeal circulation (ECC) and cardiac arrest. Anesthesia was maintained smoothly with modified NLA under normothermia. Without use of ECC, which often disturbs water and electrolyte metabolism, the patient's physical status remained quite stable. Though the left anterior descending artery was clamped for 20 minutes during the anastomosis, there was no complication, including perioperative myocardial infarction. For high risk patients with ECC such as patients with renal failure, respiratory failure and cerebrovascular diseases, this method is useful for the coronary artery bypass surgery. PMID- 8230710 TI - [A manually operated cardiopulmonary bypass circuit with two self-inflating reservoir-pumps]. AB - We improved our manually operated cardiopulmonary bypass circuit by adding another reservoir-pump in parallel to continue the venous blood withdrawal. The two reservoir-pumps were alternately and maximally compressed by hand, and hydrodynamics of the device was examined in vitro by using a physiological saline solution. When a 21 Fr, 52 cm catheter for venous drainage and a 17 Fr, 17 cm catheter for arterial infusion were used, the bypass flow rate was about 2.1 l.min-1 even under no waterhead. It was further increased to about 3.1 l.min-1 by adding a head of 60 cmH2O. The upper body of a patient undergoing resuscitation is occupied by other personnel performing mandatory procedures such as cardiac massage and endotracheal intubation. Therefore, the catheter insertion site is limited to the femoral vessels, and a long arterial return catheter will be necessary for a better cerebral oxygen supply. When a 17 Fr long arterial return catheter with 52 cm in length was used, the maximal flow rate achieved about 2.8 l.min-1 under a head of 60 cmH2O, though slightly smaller than that with the 17 cm catheter. This double reservoir-pump circuit with a long arterial return catheter will facilitate emergency cardiopulmonary bypass during acute critical resuscitation and much improve the perfusion to the vital organs. PMID- 8230711 TI - [Power spectrum of experimental gas embolism in dogs and a new device for its detection]. AB - We studied the power spectrum of precordial Doppler sounds during experimental CO2 and air embolism and developed a new device for detection (intravascular gas detector) and examined its usefulness in experimental CO2 and air embolism in dogs and in a patient during laparoscopic surgery. We did digital tape recordings during CO2 and air embolism in seven anesthetized mongrel dogs (7-16 kg body) to generate power spectrum of the heart sound on a dynamic signal analyzer. The power spectrum of "mill-wheel murmur" was found to be 10-30 dB higher than the baseline heart sound in the range of 200 and 2500 Hz. The intravascular gas detector was designed to set off alarm sounds whenever the difference of energy between the Doppler sounds low-pass filtered at 250 Hz and the Doppler sounds high-pass filtered at 250 Hz was increased. The alarm was activated immediately during experimental CO2 and air embolism and also during laparoscopic appendectomy. The alarm sounds during the appendectomy coincided with gas bubbles detected by a transesophageal echocardiography. By use of this detector, CO2 and air embolism can be easily detected and the anesthesiologist does not need to pay continuous attention to Doppler sounds by diverting his or her concentration on the patient. PMID- 8230712 TI - [The 40th general meeting of the Japanese Society of Anesthesiology. Morioko City, Iwate, Japan, April 22-24, 1993. Abstracts]. PMID- 8230713 TI - [The spread of spinal anesthesia in young patients]. AB - We have studied the spread of spinal anesthesia with Neo-percamine S, containing hyperbaric dibucaine, in 20 younger patients (13-18 yr) and 20 older patients (22 61 yr). In younger patients, 2.17 +/- 1.7 (mean +/- SD) ml of the anesthetic solution blocked 19.4 +/- 1.7 spinal segments. In older patients, 2.22 +/- 0.18 ml of the solution blocked 16.3 +/- 0.18 spinal segments. Spinal anesthesia higher than T3 level was observed in 45% of the younger patients, whereas such level of spinal anesthesia was not observed in the older patients. Our results suggest that hyperbaric dibucaine solution for spinal anesthesia may have a tendency to produce an unexpectedly extensive spread of anesthesia in young patients. PMID- 8230714 TI - [The relation of vertebral artery blood flow velocity to mean arterial pressure during anesthesia with enflurane or isoflurane]. AB - The relation of vertebral artery blood flow velocity (VABFV) to mean arterial pressure (MAP) during anesthesia with enflurane or isoflurane was examined. The subjects were 17 patients on the knee-chest position. Anesthesia was induced with thiamylal and fentanyl. The patients were placed on the knee-chest position after tracheal intubation. Ventilation was adjusted to maintain a constant level of PaCO2 (30-40 mmHg). The patients were divided into two groups. Anesthesia was maintained with enflurane and 67% N2O in group E (8 patients), or with isoflurane and 67% N2O in group I (9 patients). VABFV was measured with a transcranial Doppler sonography (TCD), and MAP was measured through a radial artery catheter. The mean VABFV and MAP were recorded at five-minute intervals. VABFV was found to change with the change in the MAP for the same degree in both groups. The results suggest that cerebral autoregulation is disturbed during either isoflurane-N2O or enflurane-N2O anesthesia. PMID- 8230715 TI - [Emergency cardiopulmonary bypass (ECPB) for cardiopulmonary-cerebral resuscitation; (2). Physiologic changes]. AB - After normothermic ventricular fibrillation (Vf) cardiac arrest of 15 min, 15 female mongrel dogs received conventional cardiopulmonary resuscitation (CPR) for 3 min and then cardiopulmonary bypass through the femoral artery and veins (F-F bypass). Cardiac beat did not return in any dogs during the initial 3-min CPR. Spontaneous circulation was restored by defibrillation 5.2 +/- 3.8 (mean +/- SD) min after the initiation of the F-F bypass in all dogs except in one with bypass trouble. Life-threatening ventricular dysrhythmias appeared in 8 of the 14 dogs (57%). Eleven of the 14 dogs (79%) were successfully weaned from the F-F bypass. Spontaneous respiration and circulation remained stable for the subsequent 1-h observation period in 8 dog (57%). The values of cardiac output measured just before weaning from the bypass and 30 to 60 min after weaning were significantly lower than those before inducing Vf (n = 5). On the electroencephalogram, intermittent burst waves reappeared 90.0 +/- 24.7 min after the initiation of resuscitation and EEG showed continuous waves 130.7 +/- 28.1 min (n = 7) after the initiation of resuscitation. The values of blood glucose, lactate and potassium 5 to 15 min after the initiation of F-F bypass were significantly higher than those before induction of Vf, while the values of hemoglobin, hematocrit, platelet and serum protein decreased significantly (n = 14). PMID- 8230716 TI - [A manually operated portable cardiopulmonary bypass circuit for emergency and its hydrokinetics]. AB - We developed a manually operated portable cardiopulmonary bypass circuit for emergency. The priming volume of the circuit is about 300 ml, including the self inflating reservoir-pump of 120 ml in capacity. The self-inflating reservoir-pump with two one-way valves can drain venous blood via the femoral vein without a water head, and return the blood into the femoral artery through a membrane lung. Bypass flow under various conditions was measured with an electromagnetic flowmeter. Pressure resistance of the artificial lung was also measured. When a DLP 21 Fr size catheter was used as a venous drainage catheter and DLP 17 Fr size for arterial infusion, bypass flow was 0.8-1.2l.min-1 without a water head for venous drainage, or 1.4-1.7l.min-1 with a 60 cm water head, respectively. One investigator could repeat squeezing the reservoir-pump for two hours under each condition. The bypass flow rates of the circuit depend upon the height of the water head and the internal diameters of the venous drainage catheter. The diameter of the arterial catheter affects the squeezing pressure, but it is overcome by manual force and has no influence on the flow rate. The lung had no significant resistance either to blood flow or insufflation gases. PMID- 8230717 TI - [Interleukin-6 and glucocorticosteroid responses to cardiopulmonary bypass]. AB - The responses of interleukin-6 (IL-6), ACTH, cortisol, WBC, CRP to cardiopulmonary bypass (CPB) were studied in 5 patients who underwent elective CABG (N = 4) and AVR (N = 1). IL-6 started to increase from 3 h after the beginning of the operation at which aortic clamp was removed and reached a peak after 4 h at which CPB was withdrawn. ACTH also reached a peak after 4 h. The increase of cortisol started from 2 h when aorta was clamped, which was earlier than that of IL-6. IL-6 and ACTH fell sharply to pre-CPB levels before first postoperative day (1 POD) while cortisol remained high postoperatively. CRP level and WBC counts were maximum at 2 POD. The results suggest that the cortisol response to CPB is not only caused by IL-6, but also by other factors, such as IL 1 and TNF. PMID- 8230718 TI - [Effects of DbcAMP on tumor necrosis factor and interleukin-1 production in human monocytes]. AB - Recent reports have shown that dibutyryl cAMP (DbcAMP) blocks endotoxin-induced lung injury. To determine whether DbcAMP suppresses the production of tumor necrosis factor (TNF) and interleukin-1 (IL-1) in human monocytes, we measured the levels of TNF and IL-1 in response to E. Coli lipopolysaccharide (40 micrograms.ml-1) in vitro. We now show that DbcAMP suppressed dose-dependently the production of TNF in human monocytes, and DbcAMP totally suppressed it at the dose above 5 x 10(-4) M. However, DbcAMP did not suppress the production of IL-1 even at the dose of 5 x 10(-3) M in human monocytes. These data suggest that the productive mechanism of IL-1 may be different from that of TNF. Further, suppression of TNF by DbcAMP may contribute to the beneficial effects in animal models of septic shock or lung injury and this may have clinical implications. PMID- 8230719 TI - [Inhibitory action of bupivacaine on cardiac contraction]. AB - We studied the effects of bupivacaine on the contractility and membrane potentials in isolated dog papillary muscle. Bupivacaine (10(-6)-10(-4) M) produced dose-dependent depression of twitch tension elicited by electrical stimulation. These inhibitory effects were greater at high frequencies of stimulation (2, 3 Hz) than at low frequencies (less than 1 Hz). Bupivacaine did not alter the resting membrane potential, but produced a reduction in Vmax of the action potentials, in a dose-dependent and reversible manner (concentrations from 10(-6) to 10(-4) M). Bupivacaine 10(-4) M often blocked the fast action potentials in normal Tyrode's solution. In high K+ (26 mM) Tyrode's solution, bupivacaine inhibited both slow action potentials and associated contractions in the presence of isoproterenol. These results suggest that low concentrations of bupivacaine decreases the contraction mainly due to Na+ channel block, whereas at higher concentration, this anesthetic may block Ca2+ channels. In addition, isoproterenol may be clinically effective in the treatment of bupivacaine cardiotoxicity due to stimulation of Ca2+ mediated slow action potentials through beta-receptors. PMID- 8230720 TI - [Culture of the irrigating fluid of the epidural space obtained during chronic epidural catheterization]. AB - The most serious complication during long-term epidural catheterization is epidural infection. Bacterial culture of the irrigating fluid of epidural space was carried out periodically in 39 patients in whom epidural catheters were inserted for a long period of time. Eight (17%) of 47 samples of epidural irrigating fluid were contaminated by the normal skin flora. All of these cases were accompanied with epidural contamination by the same organisms. No significant correlation was found between clinical signs of infection (low grade fever, leucocytosis and localized infective signs at the puncture site) and the contamination of the epidural irrigating fluid or that of the epidural catheter. When epidural irrigating fluid was contaminated, the epidural catheter was removed immediately and the patient was treated by antibiotics. None of the patients had epidural abscess or neurological deficit. In conclusion, bacterial culture of epidural irrigating fluid is valuable for the early diagnosis of epidural infection during long-term epidural catheterization. PMID- 8230721 TI - [Effects of the volume of the reservoir bag and the amount of fresh gas flow on FRC during CPAP]. AB - We have examined effects of the volume of the reservoir bag and the amount of fresh gas flow (FGF) on FRC during CPAP with PEEP level of 8 or 11 cmH2O. We used reservoir bags of 4 different volumes (5, 10, 15, 20l). We set up the air way pressure graduation between inspiration and expiration (delta P) by adjusting FGF rate, then we measured delta FRC and other respiratory parameters in 8 volunteers (26.3 +/- 0.8 yo). With the same PEEP level, delta P decreased but FRC increased by increasing FGF rate. When we used a lager reservoir bag, FGF rates which are necessary to set up the minimum delta P decreased during CPAP. We could obtain a larger delta FRC by using a larger reservoir bag compare with a smaller one, at the same delta P using same CPAP level. We should consider not only delta P but also the volume of the reservoir bag to obtain a reasonable and comfortable CPAP. PMID- 8230722 TI - [Cervical epidural morphine and buprenorphine for postoperative pain relief after thoracic surgery--a comparative study in the same patients]. AB - The effects of cervical epidural morphine and buprenorphine on postoperative pain were studied in 12 patients who received thoracic surgery twice. The patients who had received morphine 3 mg on the first operation were given buprenorphine 0.15 mg on the second operation, and the others received them vice versa. Morphine or buprenorphine was administered with 6 ml of 0.25% bupivacaine before skin incision. The duration of pain relief with morphine was longer (21.5 +/- 4.0 h) than with buprenorphine (13.9 +/- 7.8 h). The result of the questionnaires to the patients after operation shows that 10 patients (84%) were satisfied with morphine whereas only 6 patients (50%) were satisfied with buprenorphine. Seven patients preferred morphine to buprenorphine for postoperative analgesia, and the rest of the patients stated that analgesic effects were similar between the two. It seems that epidural morphine 3 mg may give more excellent postoperative pain relief after the thoracic surgery than epidural buprenorphine 0.15 mg. PMID- 8230723 TI - [Herpes zoster and malignancy]. AB - A retrospective survey was conducted regarding the relationship between hospitalized patients with herpes zoster and malignancy based on clinical records. A total of 220 patient were hospitalized for treatment of herpes zoster during the past 15 years, and 23 of them had involvement with malignant tumors. Malignancy was found during hospitalization in 4 cases (1.8%). This was significantly higher than 0.27, the predicted number of malignancy cases. Three of the 4 cases were gastric cancer. The discovery rate of gastric cancer through screening at admittance was 2.4%, which was higher than 0.14%, the discovery rate in gastric cancer checkups in Hokkaido. Examination for malignant tumors should be required for all patients with herpes zoster who need to be hospitalized. PMID- 8230724 TI - [Evaluation of cross circulation in conjoined twins]. AB - This report describes the evaluation of cross circulation in conjoined twins. Female thoracoomphalopagus conjoined twins were delivered by cesarean section after 37 week's gestation. CT, MRI and echography were performed. A partial communication of pericardium and sternum, and a union of the liver were found. For the preoperative evaluation of cross circulation, a bolus of indigo carmine was injected, and the pigment appeared in the urine of the other twin. RI angiography showed that radionuclides in one twin were similar to those in the other after 5-10 minutes. During the operation, an injected bolus of SCC to one twin was not effective for the other twin. Inhaled isoflurane in one twin did not appear in the expired gas of the other twin. After the intentional hemorrhage for 3 minutes from one twin, the hemoglobin concentration of one of the twin was the same as that of the other. This demonstrates that acute hemorrhage from one twin seems to result in a significant and rapid decrease of hemoglobin in both babies equally. However, intravenous infusion of drugs to one of the twins requires a relatively long time to take effect in the other baby. We must be careful in anesthetic management of the surgical separation of conjoined twins. PMID- 8230725 TI - [Anesthetic management of an infant with Menkes disease]. AB - Menkes disease is a rare, sex-linked recessive disorder characterized by kinky hair, convulsion, mental retardation, bone and connective tissue lesions, and hypothermia. These symptoms have been attributed to suppression of copper dependent enzymes resulting from copper deficiency. We report a case of a 7-month old infant with Menkes disease who underwent repair of inguinal hernia. Anesthesia was maintained with sevoflurane-N2O-O2, and the operation was carried out uneventfully. Although the patient had been medicated with anticonvulsants preoperatively, transient seizure occurred in the recovery room. We also discuss pathophysiology and anesthetic management of a patient with Menkes disease. PMID- 8230726 TI - [A case of right phrenic nerve paralysis as a complication of internal jugular vein cannulation by anterior approach]. AB - We report a case of right phrenic nerve paralysis caused accidentally by compression after internal carotid artery puncture. Postoperatively she received respiratory management and conservative treatment after extubation. Cases of phrenic nerve paralysis following cardiac surgery have been reported, but paralysis as a complication of internal jugular vein cannulation by anterior approach is rare. PMID- 8230727 TI - [Rupture of asymptomatic mycotic aneurysm after valve replacement in infective endocarditis]. AB - Mycotic cerebral aneurysms (MCA) are one of the most serious complications of infective endocarditis. The rupture of MCA in patients under anticoagulant therapy following valve replacement carries high mortality. We encountered this serious complication in a patient who had no neurologic symptoms. A 12-year-old girl was scheduled for mitral valve replacement (MVR) 5 weeks after antibiotic therapy for infective endocarditis caused by Staphylococcus aureus. Before the surgery, she did not have any neurologic symptoms or abnormal findings in CT scanning examination. The surgery to remove her mitral valve with bacterial vegetations and replace it with an artificial valve proceeded smoothly and she appeared to begin an uneventful postoperative recovery. However, she suddenly began to complain of severe headache and became unconscious on the fifth days after MVR. A CT scan showed cerebral herniation due to a major subdural hematoma. A ruptured MCA was detected in the orbito-frontal artery and clipped in an emergency operation. She was transferred to the intensive care unit and given continuous infusion of barbiturate to prevent increase of her intracranial pressure. CT scanning and arteriography 10 days after the MCA clipping, revealed a new subdural hematoma and MCA just proximal to the previous clip. It is important to bear in mind that patients with infective endocarditis can have mycotic cerebral aneurysms without any clinical neurologic symptoms. PMID- 8230728 TI - [Anesthetic management of a patient with hypertrophic obstructive cardiomyopathy under epidural anesthesia with high dose fentanyl]. AB - Hypertrophic obstructive cardiomyopathy (HOCM) is characterized by asymmetric hypertrophy of the intraventricular septum, causing intermittent obstruction of the left ventricular outflow tract. General principles of management for HOCM include maintenance of adequate preload and afterload, and avoidance of tachycardia and increased myocardial contractility. A 57 year old male for subtotal gastrectomy and gastrojejunostomy received epidural anesthesia with epidural high dose fentanyl (0.5 mg) and 0.5% isoflurane with N2O and oxygen. Blood pressure and heart rate decreased mildly but were stable during the operation and during the recovery period from anesthesia without any complaints of pain. The results suggest that epidural high dose fentanyl anesthesia is a good method for patients with HOCM. PMID- 8230729 TI - [Rupture of renal arterial aneurysm in a pregnant patient]. AB - The author experienced a case of ruptured renal arterial aneurysm diagnosed at 32 weeks of gestation. The patient was a 32-year-old healthy woman. On admission she and her fetus were in pre-shock state. Emergency caesarean section was performed under spinal anesthesia. Postoperative CT scanning revealed the ruptured right renal artery aneurysm and hematoma around the right renal hilus. About 7 hours after caesarean section, laparotomy was performed for right nephrectomy and subtotal hysterectomy. Blood loss during operation was about 8000 ml, but postoperative course was good and she and her baby had no complications. It is suggested that arterial wall of pregnant woman has weakness due to morphological and biochemical alterations following hormonal change. Pregnant uterine compresses and dislodges abdominal aorta and inferior vena cava, and these vessel moves in a curve in a leftword and dorsal direction. Plasma volume increases in late pregnancy, therefore intra-arterial pressure is elevated. It is proposed that alterations of vessel wall, increases of circulatory blood volume and vessel displacement are implicated to the etiology of renal arterial aneurysm of pregnant woman. Pregnancy-related renal arterial aneurysm has a high risk of rupture and it is advisable to think of ruptured renal arterial aneurysm as one of the causes of hemorrhagic shock during pregnancy. PMID- 8230730 TI - [Spinal anesthesia for empty sella syndrome associated with diabetes insipidus--a case report]. AB - A 60-year-old woman was admitted to our hospital for surgical treatment of the left inguinal hernia. She had suffered from diabetes insipidus for ten years, and hormonal study revealed low plasma level of vasopressin. She has been taking nasally desmopressin acetate 5 micrograms twice a day and urinary output has been well controlled around 1200-1400 ml.day-1. CT-scan showed empty sella without any pituitary tumors. There were no evidences of increased intracranial pressure and neurological deficit. Following nasal instillation of desmopressin acetate 5 micrograms one hour before anesthesia, spinal anesthesia was performed with tetracaine 10 mg. Cephalad sensory block assessed by pinprick spread to T6 within 10 minutes. Systolic blood pressure gradually decreased from 120 to 90 mmHg, although no vasoconstrictors were needed. Arterial blood pressure was stable during the surgery. The operation lasted 80 minutes with 650 ml of fluid replacement, blood loss of 50 g and urinary output of 25 ml. She had no postspinal headache nor neurological deficit after surgery. Empty sella syndrome associated with diabetes insipidus is rare. Low spinal anesthesia can be performed safely whenever there is no evidence of increased intracranial pressure, although care should be taken for perioperative fluid and circulatory management. PMID- 8230731 TI - [A new leak test of anesthetic machine]. AB - A case of the inspired PCO2 increase secondary to the malfunction of expiratory valve due to a defected edge of creator is presented. In this case, we could not detect the malfunction of expiratory valve using the preanesthetic routine leak test but found it by observing the expiratory phase of the capnometric curve. Therefore, we devised a new leak test for checking the malfunction of inspiratory and expiratory valves. This test is simple and effective. The test consists of the following procedures.: 1. Connecting yokes to the oxygen piping system. 2. Connecting the breathing bag. 3. Occluding the inspiratory terminal. Closing the semiclosed valve completely. 4. Flushing the O2 gas and increasing the pressure of breathing circuit to 30 cmH2O. 5. Monitoring the pressure gauge of the anesthesia system. If the function of valves is normal, the system pressure decreases slowly. If either of the valves works wrong, it decreases rapidly. We checked anesthesia machines in our facilities using the new leak test, and found some cases of the expiratory valve malfunctions. PMID- 8230732 TI - [Max Kappis, an inventor of splanchnic nerve block]. PMID- 8230733 TI - [Clinical evaluation and pharmacology of DQ-2556 in the patients with compromised renal function]. AB - We studied the pharmacokinetics of a new cephem antibiotic, DQ-2556, in patients with impaired kidney function. The peak concentrations of the compound in the serum were observed irrespective of the degree of kidney failure 5 minutes after its bolus administration of 1.0 g intravenously, and no significant difference was observed in the concentrations among the patients. On the other hand, the decrease in its concentrations in the serum was impeded in proportion to degrees of kidney failure and, in particular, hemodialysis patients showed markedly delayed clearance of the drug from the serum; the half-lives in the serum (beta phase) were prolonged to ca. 6 hours in patients with severe kidney failure (Ccr ca. 20 ml/min) and did so markedly to ca. 17 to 21 hours in patients with hemodialysis as compared with ca. 2.5 hours in patients with slight kidney failure (Ccr ca. 50 ml/min). Urinary excretion rates (0-to-24 hours values) were ca. 70% in patients with slight kidney failure, ca. 60% in patients with moderate kidney failure and ca. 40% in patients with severe kidney failure, showing a tendency toward a decline in relation to increasing degrees of kidney failure. The compound showed a satisfactory dialytic property. The clinical efficacy and safety of DQ-2556 were evaluated upon administering if at daily doses of 0.5 g b.i.d. and 1.0 g b.i.d. for 7 and 14 consecutive days respectively, in patients with lower respiratory tract infections. The clinical efficacies were excellent in 2 patients, good in 11 and poor in 2, yielding a efficacy rate of 86.7%. No side effects were observed, though, a neutrophil sedimentation ratio decreased in a patient, and a down-shift of prothrombin activities was observed in another. These results suggest that DQ-2556 is useful for lower respiratory tract infections, but in patients with kidney failures it is required to seek the most suitable regimen since the excretion rates of the compound decrease as degrees of kidney failure become severer. PMID- 8230734 TI - [In vitro susceptibility to fluconazole of fungal strains freshly isolated from child patients with deep-seated mycoses]. AB - The susceptibility to fluconazole (FLCZ) were evaluated for 40 strains of 8 yeast species and 4 strains of 3 Aspergillus species, all of which were isolated from clinical specimens during clinical trials of FLCZ in child patients performed from January 1991 to January 1993 in this country. The in vitro activity of FLCZ against yeast isolates and Aspergillus isolates were measured using the microdilution method with semisolid SAAMF (synthetic amino acid medium fungal) agar and the macrodilution method with Eagle MEM (minimum essential medium), respectively, using amphotericin B (AMPH) and flucytosine as the reference drugs. Out of all yeast species tested, Candida albicans and Candida tropicalis were the most susceptible to FLCZ with geometric mean (GM)-MICs of < 0.4 microgram/ml that were far less than the corresponding values for AMPH or flucytosine. On the other hand, FLCZ showed the lowest activity against Candida krusei and Candida glabrata with GM-MICs of approximately 10 micrograms/ml that were slightly higher than those for AMPH. The susceptibilities to FLCZ of Candida parapsilosis, Candida pelliculosa (anamorph of Hansenula anomala), Candida famata, and Trichosporon cutaneum were intermediate. Antifungal activities of FLCZ and the 2 reference drugs against aspergilli were determined on the basis of the amount of total protein recovered from drug-treated cultures relative to that from drug-free control cultures. Fifty percent and 90% inhibitory concentrations of FLCZ against 2 isolates of Aspergillus fumigatus were 6.25-25 micrograms/ml and 50 micrograms/ml, respectively. Against this and 2 other species of Aspergillus, AMPH appeared more highly active than FLCZ. PMID- 8230735 TI - [A clinical study of fluconazole-granules and -injectable in pediatric patients with deep-seated mycoses]. AB - Fluconazole (FLCZ) is an antifungal agent of triazole class developed by Pfizer, Inc. Its oral and injectable forms have been available on the market since June 1989 in Japan. FLCZ exhibits potent antifungal activities against Candida spp., Aspergillus spp. and Cryptococcus spp. and, as orally or intravenously administered, is widely distributed into organs and tissues. For its low protein binding rate of about 10 per cent and long serum half life of about thirty hours in adults, FLCZ has been proved highly effective and useful in the treatment of deep-seated mycosis in adult patients. In the present study, we have investigated the clinical effectiveness and antifungal activities of FLCZ granules, a new dosage form of the drug, and of intravenous form in pediatric patients with deep mycosis. A total of 72 patients were treated either with granules orally or with intravenous injection and 47 patients among them were evaluable on the clinical efficacy of the drug. Also, a study on the pharmacokinetics of pediatric patients including premature/new born babies was conducted employing multiple dose regimens in a total of 27 patients. The clinical efficacy rates were 79.5% (35 patients out of 44) in candidiasis and 100.0% (3 of 3) in aspergillosis. The safety of the drug was assessed in 63 patients. No side effects were observed. Clinical laboratory test abnormalities were observed in some patients with an incidence of 9.7% (6 patients out of 62) but most of the abnormalities were only mild and transient. The pharmacokinetics at repeated doses indicated that a steady-state is reached in 4 days after the initial administration of either granules or intravenous form. From these results, it may be concluded that FLCZ is a very useful medication in the treatment of deep mycosis in pediatric patients. PMID- 8230736 TI - [Clinical effects of a combination treatment with cefodizime and minocycline for infections in patients complicated with hematological disorders]. AB - We evaluated clinical effects and toxicities of a combination in treatment with cefodizime (CDZM) and minocycline (MINO) for infections complicated with hematological disorders in 67 patients. Fifty-nine patients were evaluable, including 32 with acute leukemia, 15 with malignant lymphoma, and 12 with other hematological disorders. Clinical efficacies were excellent in 17 cases, good in 24 cases, fair in 2 cases, and poor in 16 cases. The efficacy rate was 69.5% (41 cases/59 cases). This treatment was also effective in 8 of 12 cases in which granulocyte counts were less than 500/microliter through the course of administration. No subjective side effects were observed. Abnormal values in laboratory tests were noted in 5 cases. Mild elevations of GOT, GPT, Al-P and bilirubin were observed, but none was serious. Thus, the combination of CDZM and MINO is an effective and safe regimen for the treatment of infections in patients complicated with hematological disorders. PMID- 8230737 TI - [Effect of combination therapy with sulbactam/cefoperazone and amikacin in the treatment of infectious diseases accompanied by hematological disorders]. AB - We studied the efficacy and safety of a combination therapy with sulbactam/cefoperazone (SBT/CPZ) and amikacin (AMK) on severe infections in the patients with hematological disorders. We administered SBT/CPZ and AMK to 65 patients and analyzed the result of 57 patients, except for 8 drop outs, for evaluation. The overall clinical efficacy of this combination therapy was 64.9% (excellent in 15 patients, good in 22 patients). Thirty seven patients had sepsis and suspected sepsis. The efficacy was 67.6% in these patients. The rate of improvement in the cases (18 patients) with neutrophil counts under 500/mm3 during the administration among 28 patients who had not responded to prior antibiotic treatment was 67.9%. Diarrhea was found in 1 patient as an apparent side effect likely related to this drug regimen. It appears that combination therapy of SBT/CPZ and AMK was a useful regimen in the treatment of infectious diseases complicated with hematological disorders. PMID- 8230738 TI - [The dose-response study of sparfloxacin against skin and soft tissue structure infections in the field of surgery]. AB - Sparfloxacin (SPFX), a new oral quinolone antimicrobial, was evaluated for the clinical efficacy against skin/soft tissue structural and osteomyelitic infections. SPFX was administered to a total of 101 patients with various infections such as infected atheroma, periproctal abscess, subcutaneous abscess, wound infections, felon, cellulitis, furuncle, pilonidal sinus, sappurative mastitis, lymphangitis, hemorrhoidal fistula, osteomyelitis. The clinical efficacy in the evaluable 101 cases was assessed by the physician in charge as excellent in 19 cases, good in 64, fair in 11 and poor in 7, the efficacy rate being 82.2%. In contrast, the clinical efficacy in 101 evaluable cases by the criteria of the committee as excellent in 36 cases, good in 45, fair in 8, and poor in 12, the efficacy rate being 80.2%. Clinical efficacy rating was not significantly difference between 200 mg/day group and 300 mg/day group. The bacteriological eradication rate was 86.5% in 53 cases with monomicrobial infection and 90.3% in 33 cases with polymicrobial infections. Of 18 cases whose infections were previously intractable with other drugs and treated thereafter with SPFX, 15 were judged in the efficacy as excellent or good. The side effects observed in 2 cases during the treatment were epigastralgia and nausea which were tolerable and did not require withdrawal of SPFX. No abnormal laboratory value was found in the several required tests. The MIC values measured for 108 strains (90.0%) of 120 clinical isolated of 35 species were lower than 0.78 microgram/ml. PMID- 8230739 TI - Lysozyme in cervical mucus of patients with chorioamnionitis. AB - Lysozyme and lactoferrin, antimicrobial substances present in cervical mucus, play an important role in defense mechanisms against ascending infections in development of chorioamnionitis (CAM). In this study, we measured the concentration of lysozyme in cervical mucus, and studied its relationship with other factors, and obtained following results. The concentration of lysozyme in cervical mucus decreased more significantly in positive CAM in preterm labor than in negative CAM and control subjects (P < 0.001). A direct correlation between the concentrations of lysozyme and lactoferrin was observed (r = 0.709). In positive CAM, the concentration of elastase in cervical mucus reached a significantly high level (more than 8,000 micrograms/L), and a no correlation between the concentrations of elastase and lysozyme was observed. These findings suggest a reduction of the defense mechanism occurred because the concentrations of both lysozyme and lactoferrin in cervical mucus during preterm labor (positive CAM) decreased. PMID- 8230740 TI - Fluctuations in elastase of cervical mucus and clinical efficacy of cefodizime in obstetric and gynecologic infections. AB - Clinical efficacy of cefodizime (CDZM) in obstetric and gynecologic infections and in their prevention, and fluctuations in cervical mucus elastase in gynecologic infections and during pregnancy were studied, and the following results were obtained: 1. Cervical mucus elastase showed high values of > or = 5,001 micrograms/L in 68.2% of non-pregnant patients and in 25.9% of pregnant patients (gestational age of 24-42 weeks). Patients with concomitant chorioamnionitis showed a significantly higher rate of elastase elevation than those without this complication (63.2% vs. 25.9%, P < 0.005). 2. Cervical mucus elastase in uterine and pelvic infections showed high values before administration of CDZM but tended to decline with the progress of cure (15/19 cases, 78.9%). 3. The efficacy rate of 2-4 g/day of CDZM drip infusion against infections was 53/56 (94.6%) while the bacterial eradication rate was 36/38 (94.7%). The efficacy rate of 2 g/day of CDZM drip infusion for the prevention of postoperative infections was 429/431 (99.8%). Diarrhea occurred in 3/489 (0.6%) as subjective and objective side effects. Slight rises in GOT and GPT were noted in 13/486 (2.7%) as abnormal laboratory values. These results suggested the usefulness of CDZM in the obstetric and gynecologic fields. PMID- 8230741 TI - [Criteria for clinical evaluation of antibiotics in pediatrics. The purpose and process for establishing the criteria]. AB - This paper describes the purpose and process for establishing "Criteria for Clinical Evaluation of Antibiotics in the Pediatric Field", which was reported in the Japanese Journal of Antibiotics Vol. 46, May, 1993. The Criteria Committee was organized in November 1991. Four meetings were held to establish the draft criteria. The criteria were applied to the evaluation of oral cephem S-1108 and parenteral cephem SCE-2787. When the criteria were compared with the conventional criteria, the results indicated that no difference was obtained in the efficacy rate as a whole, the sum of "Good" and "Excellent" cases, but there was a difference in the cases judged to be "Excellent". Partial alteration was made to the draft criteria and the Committee produced the final version of the criteria. However, the criteria are far from complete, so it will be subjected to further revision it accordance with future advance in chemotherapy. PMID- 8230742 TI - [Effects of rHuEPO on aplastic anemia: results of a phase II clinical study]. AB - The safety and efficacy of recombinant human erythropoietin (epoetin alpha) were investigated in adult aplastic anemia patients whose hemoglobin (Hb) concentration was less than 10g/dl. Epoetin alpha was given subcutaneously every day at a dose of 3,000IU/body 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. In cases in whom Hb concentration increased by more than 1g/dl or whose transfusion requirements reduced to less than 50%, treatment was judged to be effective. The whole rate of efficacy was 34.5% (10/29). Response to epoetin alpha treatment was better in patients whose symptoms were relatively mild. Mild cases responded to the treatment with 6,000IU/body/day, although a dosage of 24,000IU/body/day was required in moderate or severe cases. Neither serious adverse effect nor abnormal laboratory findings were observed. These results suggest that high dose subcutaneous epoetin alpha treatment is effective for the aplastic anemia in terms of increasing Hb concentration and reducing blood transfusions. PMID- 8230743 TI - [Non-Hodgkin's T cell lymphoma associated with marked eosinophilia]. AB - A case of non-Hodgkin's T cell lymphoma (diffuse lymphoma, large cell type) associated with marked eosinophilia and pleurisy in a 57-year-old male is reported. The leukocyte count was 12.5 x 10(3)/microliters and eosinophil count was 53% and the absolute count of 6.6 x 10(3)/microliters. The patient's serum and pleural effusion fluid, containing abundant lymphoma cells, showed eosinophil colony stimulating factor (Eo-CSF) activity. Conditioned medium (CM) prepared from patient's T cells (T-CM) produced Eo-CSF and this was enhanced by interleukin-2 (IL-2) stimulation. We demonstrated that the patient's serum contained a significant amount of interleukin-5 (IL-5) and the patient's T-CM, particularly after IL-2 stimulation contained a significant amount of granulocyte macrophage colony-stimulating factor (GM-CSF). These findings suggest that Eo-CSF produced by neoplastic T cells or normal T cells activated by tumor antigen stimulated the production of eosinophils in this patient and that both IL-5 and GM-CSF might play a role in Eo-CSF activity. PMID- 8230744 TI - [B cell malignant lymphoma complicated with partial Addison's disease, report of a case]. AB - A 58-year-old man was admitted in May 1988, because of high fever, skin, pigmentation and body weight loss. Abdominal ultrasonography and generalized computed tomography examinations showed swelling of general lymph nodes and bilateral adrenal glands, splenomegaly, and lesion in the liver. Serum cortisol, urinary 17-OHCS and 17-KS level were within the normal range, while the ACTH level was elevated (189.9 pg/ml). ACTH overload test showed a non-reactive pattern, leading to a diagnosis of partial Addison's disease. He was also diagnosed as non-Hodgkin lymphoma, diffuse, large cell type (B) by a biopsy of the left supraclavicular lymph node. After combination chemotherapy, swelling of the supraclavicular lymph node diminished, followed by normalization of ACTH level and improvement of symptoms. Abdominal lymphadenopathy, hepatosplenomegaly and swelling of adrenal glands also decreased in size. Although involvement of tumor cells in adrenal glands sometimes occurs in malignant lymphoma, it is reported that more than 90% destruction of adrenal gland tissue is necessary to develop Addison's disease. It was suggested that the involvement of many lymphoma cells in both adrenal glands resulted in the development of partial Addison's disease in this case. PMID- 8230745 TI - [Improvement of anemia by recombinant human erythropoietin in paroxysmal nocturnal hemoglobinuria]. AB - A 32-year-old man visited Kanto Teishin Hospital complaining of general fatigue in May, 1992. He had been diagnosed as having paroxysmal nocturnal hemoglobinuria since 1980, because of brownish urine in the morning. He received blood transfusion in 1980. In 1983, he was treated with medication. There was no remarkable improvement, however, and he stopped coming to the hospital. When he was admitted to our hospital, hemolytic anemia and hemosiderinuria were noticed. Sucrose hemolysis test and acidified-serum lysis test (Ham test) were both positive. Positive rates of decay accelerating factor and CD59 were 38.8% (control 100%) and 45.4% (control 100%), respectively. His diagnosis was thus confirmed. Bone marrow was slightly hypocellular, and erythroid cells were relatively hyperplastic (M/E ratio 0.68). The oral administration of iron and oxymetholone was not effective for anemia. He was treated with daily subcutaneous administration of recombinant human erythropoietin (EPO, 3,000U/body/day). His hemoglobin level increased from 7.5g/dl to 12.0g/dl in 4 weeks, and general fatigue disappeared. Since he had concurrent chronic hepatitis C, alpha interferon was also administered and his hemoglobin level is now controlled between 10 and 11g/dl. This case suggests that EPO can be useful for treating hemolytic anemia, even though erythroid cells in the bone marrow are hyperplastic. PMID- 8230746 TI - [Sideroblastic anemia preceded by essential thrombocythemia with 20q- chromosome abnormality]. AB - A 78-year-old man presented with marked thrombocytosis (126.4 x 10(4)/microliters), low neutrophil alkaline phosphatase (NAP) score and an abnormal karyotype of 46, XY, del(20) (q11q13) (18 of 20 cells), without obvious anemia or ringed sideroblasts in bone marrow. He received ranimustine (MCNU) with a diagnosis of essential thrombocythemia. After 2 years, he was admitted because of macrocytic anemia. The peripheral blood smear showed anisopoikilocytosis with a few nucleated red blood cells. Moderate thrombocytosis (71.7 x 10(4)/microliters) and a low NAP score were also observed. Bone marrow aspiration revealed erythroid hyperplasia with a significant increase in ringed sideroblasts (85% of erythroblasts). Cytogenetic studies showed the same abnormal karyotype 46, XY, del(20) (q11q13) in 100% of metaphase cells as those at initial diagnosis. A diagnosis of sideroblastic anemia preceded by essential thrombocythemia was made. No rearrangement or amplification of c-src was revealed. The observation of the same chromosome abnormality (20q-) in different phases of this patient's disease indicates that chronic myeloproliferative disorders and myelodysplastic syndrome may share some borderline or transitional cases with a similar pathogenesis. PMID- 8230747 TI - [Effective methyl prednisolone pulse therapy for a patient with retinoic acid syndrome in acute promyelocytic leukemia]. AB - A 46-year-old woman with acute promyelocytic leukemia (APL) was treated with all trans retinoic acid (ATRA) and chemotherapy according to the AML-92, M3 regimen of the Japan Adult Leukemia Study Group (JALSG). Between days 7 and 18 of therapy, she suffered chest discomfort, fever, cough, dyspnea and general fatigue. A chest roentogenogram showed bilateral interstitial infiltrates. Her leukocyte count began to increase rapidly to 6,400/microliters on day 14. Marked hypoxia (PO2 35.9 mmHg) suggested occurrence of retinoic acid (RA) syndrome. She underwent endotracheal intubation and mechanical ventilation with administration of methyl-prednisolone (m-PSL) pulse therapy. Her symptoms promptly abated. Therapy with ATRA was continued and her leukocyte count reached 44,800/microliters on day 19 of therapy. She achieved complete remission on day 48. PMID- 8230748 TI - [Primary myelofibrosis in an infant--a case report and review of the literature]. AB - Primary myelofibrosis (PMF) is regarded as a chronic myeloproliferative disorder. It is characterized by marrow fibrosis, leukoerythroblastosis, tear drop erythrocytes and extramedullary hematopoiesis. Most patients are in their late 50s when first diagnosed. Pediatric PMF is said to be quite rare. Here describe a female infant with PMF. The patient was born on Aug. 7, 1991. The pregnancy and delivery were uneventful. Hepatomegaly was noted soon after birth. Combined blood counts showed polycythemia and leukocytosis. It was thought to be extramedullary hematopoiesis due to intrauterine infection. She was followed up in another hospital, but since her condition was unchanged she was admitted to our hospital for further medical examinations at age 7 months. On the peripheral blood smear, there were tear drop erythrocytes, normoblasts and early myeloid elements. Repeated bone marrow aspirations were dry taps. This case presented the classical findings of fibrosis of the bone marrow on bone marrow biopsy. She is in good health without any therapy until now. A review of 7 cases of PMF, including our case, in Japanese children was made and discussed in comparison to adult cases. PMID- 8230749 TI - [A "retinoic acid syndrome" observed in two cases of acute promyelocytic leukemia]. AB - Two cases of acute promyelocytic leukemia (APL) treated with all-trans retinoic acid (ATRA) developed fever, dyspnea and chest pain. A chest roentgenogram showed bilateral pleural effusion (case 1) and bilateral interstitial infiltration (case 2). The first case was a 50-year-old female in her first relapse, who was initially diagnosed as having pleuritis tuberculosa and was treated with anti tuberculotic agents. Her symptoms continued for 44 days and complete remission was achieved 53 days after commencing ATRA therapy. The second case was a previously untreated 46-year-old male. His case had been diagnosed as adult respiratory distress syndrome and he had been treated with prednisolone. His symptoms rapidly improved and complete remission was achieved 38 days after the ATRA therapy. This was the first report of patients in Japan considered to have developed "retinoic acid syndrome (RAS)". In our five APL cases treated with ATRA, the syndrome was not always accompanied by peripheral blood leukocytosis even though the two cases with RAS showed higher leukocyte counts than the other two cases without RAS and also had DIC. We should pay attention to the severe respiratory symptoms that develop in APL patients after ATRA treatment and immediate steroid therapy is required for such patients. PMID- 8230750 TI - [Bullous amyloidosis]. AB - Although bullous amyloid lesions are very rare, the cutaneous lesions of this type can be a crucial manifestation of plasma cell dyscrasia. [Case Report] A 66 year-old man with a six-year history of multiple myeloma (IgG-lambda, lambda-type Bence Jones proteins) was admitted to the hospital because of hemorrhagic bullous lesions of the skin, chronic diarrhea and general malaise. A diagnosis of myeloma associated amyloidosis with renal failure was made. One month later, he died as a consequence of progressive renal failure and systemic amyloidosis. A postmortem examination revealed myelomatous infiltrations (bone marrow and kidneys) and widespread amyloid deposits (skin, heart, lungs, kidneys, liver and intestine). The histologic examination of a bullous lesion showed amyloid deposits with formation of an intradermal blister. PMID- 8230751 TI - [Clinical characteristics and treatment results of acute promyelocytic leukemia in children (Children's Cancer and Leukemia Study Group)]. AB - The clinical characteristics and treatment outcome in 40 children with acute promyelocytic leukemia (APL) treated at institutions participating in the Children's Cancer and Leukemia Study Group (CCLSG) were studied retrospectively. The median age at diagnosis was 8 years old. Bleeding diathesis was the predominant presenting symptom (90%), associated with laboratory findings of disseminated intravascular coagulation. Hepatomegaly, splenomegaly and lymphadenopathy were observed in 35%, 10%, and 15% of the cases, respectively. The median WBC count was 4.25 x 10(9)/l. Anemia (hemoglobin < 8 g/dl) and thrombocytopenia (< 30 x 10(9)/l) were present in more than half of the patients. Cytogenetic studies demonstrated the characteristic 15; 17 translocation in about 90% of the patients analyzed. Induction therapy consisted of cytosine arabinoside and an anthracycline, with or without other agents. Twenty-nine patients (73%) achieved complete remission (CR) while early fatal hemorrhage was the predominant cause of induction failure. The survival rates continued to decrease (28% at 3 years, 24% at 5 years, and 7.9% at 10 years) due to late marrow relapses. Anthracycline cardiotoxicity was fatal in three patients in remission. These clinical features of childhood APL should be taken into account in the development of new protocols. PMID- 8230752 TI - Endometrium-myometrium ratio: a newly proposed diagnostic parameter on magnetic resonance imaging assessment of myometrial invasion by endometrial cancer. AB - In order to improve the accuracy of magnetic resonance (MR) imaging assessment of myometrial invasion by endometrial cancer, the usefulness of a new diagnostic parameter, the endometrium-myometrium (EM) ratio has been evaluated. EM ratio is the proportion of the widest length of endometrium to the length of myometrium measured at the same line, this being vertical to the parallel of the long axis of the uterine body in the sagittal plane of the MR images. Myometrial invasion was defined as a value of the EM ratio > 1, and the tumor was limited to the endometrium for values < 1. In 25 consecutive patients, both the EM ratio-based assessment and the well-established junctional zone-based assessment with T2 weighted MR imaging and enhanced MR imaging with gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA) were compared with the results from pathological examinations of postoperative specimens. In identifying myometrial invasion by endometrial cancer, the sensitivity of the EM ratio-based assessment was better than that of the junctional zone-based assessment. The overall sensitivity of the former was 96% in both the T2-weighted and enhanced MR imaging with Gd-DTPA, whereas that of the latter was 84% in the T2-weighted MR imaging and 72% (P < 0.05) in the enhanced MR imaging. The use of the EM ratio with MR imaging improves the ability to assess myometrial invasion by endometrial cancer. PMID- 8230753 TI - Pharmacokinetic re-evaluation and phase I study of high dose epirubicin in advanced non-small cell lung cancer. AB - We performed a phase I trial to evaluate the toxicity and the maximum tolerated dose of high dose epirubicin on a three-consecutive-day schedule on Japanese patients with advanced non-small cell lung cancer. Fourteen patients were entered in the study. At least three patients were assigned to each different dose level. Epirubicin was given intravenously daily for three day by bolus injection. The dose was started at 60 mg/m2/course and escalated by 30 mg/m2/course. Granulocytopenia was found to be the dose limiting toxicity with a maximum tolerated dose of 150 mg/m2/course. Thrombocytopenia and non-hematological toxicities were mild and well tolerated. The maximum tolerated dose was lower than that in Europe and Canada. Partial responses were observed in two out of five patients on 150 mg/m2/course. The recommended phase II dose for high dose epirubicin was demonstrated to be 120 mg/m2/course. A further dose-escalating study of epirubicin in conjunction with the administration of granulocyte colony stimulating factor is scheduled for the determination of its antitumor activity in non-small cell lung cancer. PMID- 8230754 TI - Anastomotic leakage after colorectal cancer surgery: a risk factor for recurrence and poor prognosis. AB - In order to discover the incidence of recurrence and prognosis of patients with anastomotic leakage after colorectal surgery, 980 colorectal cancer patients who underwent anastomosis at Keio University Hospital between 1970 and 1990 were examined. Thirty-three patients (leakage group) out of the 980 exhibited anastomotic leakage. The incidence of local recurrence in the leakage group was significantly higher than in the no leakage group (P < 0.01). The disease-free survival rate of the leakage group was significantly lower than that of the no leakage group in Dukes' A, B patients (P < 0.01), but was not so in Dukes' C, D patients. These results suggested that anastomotic leakage after colorectal cancer surgery might enhance the incidence of local recurrence and make the prognosis poor. PMID- 8230755 TI - Transitional cell carcinoma of the upper urinary tract: analysis of morphology and distribution for surgical management. AB - Two series of transitional cell carcinomas (TCC), one including 49 renal pelvic and/or ureteral cancers and the other 29 bladder cancers with concomitant ureteral involvement, were reviewed to establish the criteria for nephron-sparing surgery and for selecting the most appropriate surgical intervention for TCCs at the ureterovesical junction. The following categorization of tumors was made possible by histological mappings of step-sectioned surgical specimens from various surgical modes: 1) low grade papillary superficial tumor, 2) high grade papillary or non-papillary tumor with adjacent or skipped carcinoma in situ (CIS), 3) high grade non-papillary invasive tumor without CIS, 4) high grade papillary superficial tumor without CIS. We can, by comparing postoperative clinical courses, select the mode of surgery in the following manner: 1) nephron sparing surgery could be indicated in cases of low grade papillary tumor so far as a complete resection is possible; 2) in a case of high grade papillary or non papillary tumor with adjacent or skipped CIS, extended resection of the urinary tract is required; 3) in a case of high grade non-papillary tumor without CIS, complete en bloc resection of the tumor site, including surrounding organs, will be necessary. The possibility of organ-sparing surgery for early stage cancer without CIS at the ureterovesical junction has been suggested. Preoperative consideration of the above classifications for conservative surgery is also discussed. PMID- 8230756 TI - Solitary metastasis from papillary thyroid carcinoma in cirrhotic liver with hepatocellular carcinoma. AB - We report a surgical case of hepatocellular carcinoma and solitary liver metastasis from papillary thyroid carcinoma in different lobes of the liver. The former, located in the right anterior superior segment, and the latter, in the left caudate lobe, were resected simultaneously. The hepatocellular carcinoma was a micro(thin)-trabecular, pseudoglandular type of Edmondson's grade II. The liver metastasis was a papillary carcinoma of follicular variant from the thyroid. This is the first report of hepatocellular carcinoma accompanying a solitary liver metastasis from papillary thyroid carcinoma without metastasis in other organs. PMID- 8230757 TI - Primary pulmonary hemangiopericytoma: a case report. AB - Primary pulmonary hemangiopericytoma is a very rare tumor. A case of the disease is reported, together with a review of the literature. The patient was a 78-year old male, who was admitted to hospital with an abnormal shadow on his chest x ray. A primary tumor, located in the left S4, was resected, and a final diagnosis of hemangiopericytoma of pulmonary origin was made. The disease recurred in the left lower lobe 14 months postoperatively. The patient received radiotherapy and is alive to date, 23 months after the operation. PMID- 8230758 TI - Malignant giant cell tumor of the tendon sheath: an autopsy report and review of the literature. AB - A case of malignant giant cell tumor of the tendon sheath of the right hip, which developed in a 72-year-old Japanese woman, is described. The tumor exhibited histological similarities to a benign giant cell tumor of the tendon sheath (localized nodular tenosynovitis). The resected tumor, measuring 9 x 9 x 11 cm, was located in the adductor muscle and invaded the proximal femur and acetabulum. The nodule was encapsulated with a thin membrane which was soft and gelatinous in consistency and varied in color from yellow to brown. The synovium of the hip joint was normal. The primary lesion was composed of plump polyhedral and spindle shaped cells. The nuclei were large, irregular and hyperchromatic, and contained prominent nucleoli. A moderated number of multinucleated giant cells was scattered throughout the lesion. There was little stromal collagen. In the majority of the specimens, pseudoglandular or alveolar spaces were predominant. An ultrastructural study demonstrated three cell types: fibroblast-like, histiocyte-like and an intermediate. The patient underwent reconstructive surgery with a Dacron fabric-enveloped alumina ceramic pelvic prosthesis and total hip components after resection of the primary lesion. Unfortunately, because of a local recurrence, a hemipelvectomy was required 10 months after the initial operation. At that time the intestines were involved with the recurrent tumor, and the patient subsequently died of perforative peritonitis. An autopsy revealed distant metastases to the right pelvis, urinary bladder, right ureter, ilium, mesenterium and lungs. PMID- 8230759 TI - A ureteral small cell carcinoma mixed with malignant mesodermal and ectodermal elements: a clinicopathological, morphological and immunohistochemical study. AB - A 60-year-old male with a small cell carcinoma of the right lower ureter is presented. The tumor mainly comprised a small cell carcinoma but also included a full variety of histological types such as transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma, leiomyosarcoma and chondrosarcoma. Immunohistochemical staining was positive for neuron specific enolase and cluster 1 small cell lung cancer antigen/N-CAM in the small cell carcinoma and S-100 in the chondrosarcoma component. The patient underwent a right nephroureterectomy, and received prophylactic radiation of the pelvic and para-aortic lymph node regions and cisplatin and etoposide combination chemotherapy. Eight months after the chemotherapy, a transitional cell carcinoma was found in the bladder neck, and a cystectomy with urethrectomy performed. To our knowledge, this is the second report of a small cell carcinoma originating from the ureter. PMID- 8230760 TI - [A fundamental concept for development of sample transportation system in clinical laboratory]. PMID- 8230761 TI - [Fundamental challenges in the development of hematology sample transport systems]. PMID- 8230762 TI - [Combination of laboratory automation system and hospital information system]. PMID- 8230763 TI - [Requirement of conveying system in clinical laboratory and supporting from hospital information system]. PMID- 8230764 TI - [Robot handling system and conveying system of hospital laboratory]. PMID- 8230765 TI - [Hematology laboratory system and algorithm for repeated analysis in cell counting]. PMID- 8230766 TI - [Introduction of a hematology laboratory system and its evaluating]. PMID- 8230767 TI - [Laboratory conveying system in Oh-ita Prefectural Hospital]. PMID- 8230768 TI - [On-line system of Toa, Hitachi hematology analyzer in application and practice]. PMID- 8230769 TI - [Present status and problems of the Hitachi fully automated sample handling system lined with chemical analyzer 7350 E and 7250]. PMID- 8230770 TI - [Is a sample handling system essential for laboratory integration?]. PMID- 8230771 TI - [Evaluation of Hitachi automatic pre-test sample processing system--effects on eradication of error in aliquoting and saving of specimen volume]. PMID- 8230772 TI - [Total turn around time by clinical chemistry laboratory system]. PMID- 8230773 TI - [The construction of the automatic sample handling system--a point of view from a manufacturer]. PMID- 8230774 TI - Effects of topical mitomycin C on glaucoma filtration surgery. AB - We studied the efficacy and safety of using topical mitomycin C (MMC) as an adjunct to glaucoma filtration surgery. Trabeculectomy was performed in 23 eyes of 19 patients with poor surgical prognosis. After the preparation of a scleral flap, 0.2 mg or 0.4 mg/ml MMC was applied to the exposed tissue for 5 minutes. The wound was then irrigated with 250ml of normal saline. The mean follow-up period was 7.8 months. Preoperative mean intraocular pressure (IOP) was 33.8mmHg, and the mean IOPs on 1, 3, 6, and 12 months after operation were 10.3, 12.5, 12.4 and 12.3mmHg, respectively. At postoperative 12 months, 74.7% achieved an IOP of less than or equal to 20mmHg without any antiglaucoma medication. There were early postoperative complications of aqueous leaking from conjunctival wounds in 3 eyes (13.0%), shallow anterior chamber in 2 eyes, and hyphema in one eye and one eye had long-term hypotony lasting more than 3 months. Although MMC is simple to use, it is a potent adjunct to glaucoma filtration surgery, more work should follow to determine the mechanism of action, indications, dosage and optimal exposure time of MMC. PMID- 8230775 TI - Effects of sodium hyaluronate on filtration bleb in rabbits. AB - Sodium hyaluronate can be used during a trabeculectomy to prevent early postoperative hypotony and shallow anterior chamber. To determine its long-term effect on the outcome of filtration surgery, fifteen rabbits underwent a posterior-lip sclerectomy in both eyes. Into their right eyes 0.2ml sodium hyaluronate, and into left eyes 0.2ml balanced salt solution were injected intracamerally. Gross and histopathological differences of bleb were observed. There was no statistically significant difference in longevity of the filtration bleb. PMID- 8230776 TI - Trephination with vacuum trephine in undercorrection of myopic epikeratoplasty. AB - For the treatment of undercorrection after myopic epikeratoplasty, early suture removal, scar revision, retrephination, replacement of lenticule and, recently, excimer laser photorefractive keratoplasty have been employed. We performed trephination with Hessburg-Barron vacuum trephine on 11 eyes of 11 patients whose post-epikeratoplasty myopic power was over -4.00 diopters for 3 consecutive months. Patients were followed up on post-trephination 1 month, 3 months and 6 months. The mean duration from epikeratoplasty to trephination was 14.27 months, the mean pre-trephination spherical equivalent was -8.50 D and the mean keratometric reading was 40.87 D. The mean reduction of spherical equivalent was 4.07, -5.99, -8.02 D at post-trephination 1, 3, 6 months, respectively. The mean keratometric reading was 37.60 diopters at 1 month and 41.53 diopters at 6 months. At 1 and 3 months, there were significant reductions of myopic power in refraction and keratometry (p < 0.01), but, at 6 months, regression to pre trephination levels took place (p > 0.05). There was no change of uncorrected and best corrected visual acuity between baseline and post-trephination 6 months. PMID- 8230777 TI - Excimer laser photorefractive keratectomy for astigmatism. AB - The effectiveness of excimer laser photorefractive keratectomy (PRK) for astigmatism was evaluated. We treated 136 eyes of naturally occurring myopic astigmatism using the 193-nm excimer laser with an expanding slit and an iris diaphragm, and followed for a minimum of 6 months. At 6 months after operation, the mean refractive cylinder decreased from 1.62 +/- 0.88 D preoperatively to 0.48 +/- 0.48 D. For detailed analysis of the results, the surgically induced refractive change (SIRC) was determined from the preoperative and postoperative refractions by vector analysis of astigmatism. When we compared the cylinder of the SIRC with the preoperative refractive cylinder, the effect of the cylindrical ablation was 93.9 +/- 36.7% at 6 months. The axial error of the cylinder of the SIRC to the preoperative cylindrical axis was 5.9 +/- 10.2 degrees at 6 months. These results suggest that excimer laser PRK with an expanding slit appears to have a significant effect for the correction of astigmatism. PMID- 8230778 TI - Treatment of corneal neovascularization with argon laser. AB - Corneal neovascularization, which is associated with complications in corneal diseases, can cause lipid deposit, decreasing vision, and graft rejection after penetrating keratoplasty (PKP). Corneal laser photocoagulation using an argon laser or yellow dye laser for ablation of corneal neovascularization has been described. We performed corneal argon laser photocoagulation (CALP) in two male patients with corneal neovascularization after herpetic keratitis. One PKP was performed after CALP and restored good vision with no rejection of the graft during an 8 month follow-up period. In the other case we observed visual improvement and no recurrence of corneal neovascularization after CALP. PMID- 8230779 TI - [Progress of geriatric medicine in Japan]. PMID- 8230780 TI - [Clinical characteristics of elderly patients]. PMID- 8230781 TI - [Myocardial infarction in the elderly]. PMID- 8230782 TI - [Percutaneous transvenous mitral commissurotomy in elderly patients with mitral stenosis]. AB - Sixteen elderly patients (age > or = 60 years, elderly group) with mitral stenosis and 20 young patients (age < or = 59 years, young group) underwent percutaneous transvenous mitral commissurotomy (PTMC) by the single balloon technique. Pulmonary arterial and mean left atrial pressures, mitral valvular pressure gradient and left atrial dimension were significantly decrease in both groups after PTMC. Mitral valve areas by Gorlin formula were significantly increased from 1.14 +/- 0.40 to 1.78 +/- 0.66 cm2 in the elderly group and from 1.25 +/- 0.50 to 2.04 +/- 0.81 cm2 in the young group after PTMC. Thirteen elderly and 19 young patients became in NYHA class I or II after PTMC, and three elderly patients with early gastric cancer had the lower risk of the gastrectomies by preoperative PTMC. But, two elderly patients remained in NYHA class III or IV due to increased mitral regurgitation and late tamponade, and one elderly patient died by heart failure after PTMC. PTMC has the advantages of shorter hospitalization and lower invasion for the patients compared with surgery, therefore PTMC is an effective treatment in both elderly and young groups. Elderly patients with severe mitral stenosis have higher risk of PTMC and are needed discrete strategy. PMID- 8230783 TI - [Feelings of well-being and depression in relation to social activity in normal elderly people]. AB - In order to clarify the influence of social activities on the feeling of well being or depression in the elderly, we studied these relationships in 2 groups with different activities. Group I consisted of 26 subjects (mean age, 77.2 years) living in a retirement house and exposed to relatively few social stimuli. Group II consisted of 47 subjects (mean age, 75.6 years) who were living in their own homes and were confirmed to be socially active. The Morale Scale and the Zung Self-Rating Depression Scale (SDS) were used to evaluate feelings of well-being or depression. The morale scale in group I was significantly lower than in group II. In particular, there was a significant difference in the factor related to aging. The SDS score of group I was significantly higher than that of group II. The depressive state incidence was significantly higher in group I than in group II in all subjects. There was a highly significant correlation between the morale scale score and the SDS score. These results indicate that group I is less satisfied and more depressed than group II. We conclude that social environmental factors are extremely important for the quality of life of elderly people. PMID- 8230784 TI - [The changes in physico-chemical parameters obtained from apparently healthy aged people followed over ten years]. AB - The author assessed the physiological aging parameters of 38 apparently healthy subjects who were over 80 years old 1989, who were not on medication, and who had consulted with the Keio Health Counseling Center over 10 years. All subjects had no history of overt vascular disease and/or malignancy in 1989. In 17 of 38 subjects, physical, hematological and blood chemical parameters when they were in their 70s were analyzed. Many parameters were unchanged and remained within normal limits for ordinary adults. Cataract, atherosclerotic change of optic fundi and diagonal ear lobe creases were seen in all subjects during the study period. Concerning standard deviations, those of forced expiratory volume in one second/predicted vital capacity and pure tone average (acoustic ability) decreased with age, unlike those of other parameters. Furthermore, multiple regression analysis, revealed that serum albumin decreased but pure tone average, Scheie's atherosclerotic score, senile cataract, HDL-cholesterol blood urea nitrogen and forced expiratory volume in one second/predicted vital capacity increased with age. This study was not cohort study with selected subjects but shows very slight change of almost any parameter irrespective of age and abnormality can suggest the existence of disease. PMID- 8230785 TI - [A case of early duodenal cancer resected by endoscopic polypectomy in the elderly]. AB - Early duodenal cancer has been detected with increasing frequency of endoscopy and the advent of an aging society. Aged patients, however, are often inoperable because of complications due to various underlying diseases. The authors report a case of early duodenal cancer curatively operated by endoscopic polypectomy and refer to related cases in the literature. PMID- 8230786 TI - [Two cases of hepatic encephalopathy associated with a high-intensity area in the basal ganglia on T1-weighted MR images]. AB - The authors report two cases of hepatic encephalopathy with chronic hepatic failure. Case 1 was a 78-year-old woman with liver cirrhosis, admitted because of general fatigue and loss of appetite. Her electroencephalogram showed frequent slow waves in the theta range with intermittent triphasic waves T1-weighted MR images showed increased signal intensity in the globus pallidus and the putamen. Case 2 was a 71-year-old woman with chronic hepatitis, admitted because of depression. Her electroencephalogram showed frequent slow wave activities in the theta-delta range with intermittent trisphasic waves. Her serum ammonia level was 84 micrograms/dl (normal 12-54 micrograms/dl). T1-weighted MR images showed increased signal intensity in the globus pallidus, the putamen and the hypothalamus. On the basis of these findings, both patients were diagnosed as having hepatic encephalopathy, although disturbance of consciousness was not obvious. The observed MR image abnormalities might be due to the metabolic and pathological changes of chronic hepatic failure. Such MRI findings may be useful for the diagnosis of hepatic encephalopathy. PMID- 8230787 TI - [A case of pancreatic pseudocyst with intracystic hemorrhage and repeated gastrointestinal bleeding]. AB - The case is a 83-year-old woman who came to our hospital with melena and epigastralgia as chief complaints, presenting pre-shock state with high serum amylase value. ERP revealed diffuse dilatation of pancreatic duct and cyst of the accessory pancreatic duct. Endoscopy showed hemorrhage of the accessory papilla. A diagnosis of chronic pancreatitis and hemosuccus pancreaticus was made. Because of the patient's advanced age, conservative treatment (total parenchymal nutrition, transfusion, hemostatics, etc.) were prescribed. The patient succumbed due to heart failure 15 months later. This case of gastrointestinal bleeding from a pancreatic pseudocyst due to neighboring arterial rupture is discussed with reference to some related literature. PMID- 8230788 TI - [The significance of repeated users of geriatric health care facility]. AB - Our geriatric health care facility was established 3 years ago. The real number of users since April 1989 to March 1991 amounts to 519, of which 62 have been admitted 3 times or more (defined as repeated here). At the same time, this facility has a day care activity 6 times a week. The utilization rate of day care activity was 14.8% for total users, while it was 59.7% for repeaters. These results indicate that repeaters actively utilize day care activity as well as geriatric health care facility. In other words, repeaters usually reside at home and their home cares are greatly supported by both repeated admissions to the facility and daily utilization of day care activity which are effective in improving ADL of the disabled elderly. It is concluded that geriatric health care facility plays an important part in supporting home care of the disabled elderly as well as transitional step from hospital to their own home. PMID- 8230789 TI - [Validity of clinical diagnosis in age-associated dementia]. AB - With neuropathological diagnosis as the point of reference, the validity of clinical diagnosis was studied in 50 patients who had met DSM-III-R criteria for dementia. Clinical diagnosis of ATD (dementia of Alzheimer type) was made in 14 cases, of VD (vascular dementia) in 14 cases, of others in 5 cases and of undefined in 16 cases. At necropsy, 16 cases fulfilled the histological criteria for ATD, while 15 cases were VD, 3 cases were mixed dementia and 14 cases were other CNS disorders. In 2 other cases, the neuropathological picture was considered to be nondiagnostic. The accuracy of clinical diagnosis of VD was higher than that of ATD. Clinical diagnostic sensitivity was 81.8% for ATD and 91.7% for VD; corresponding specificity was 78.3% and 86.4%. With our clinical criteria, slowly evolving VD with no clinical evidence of stroke would not be identified as VD. On the other hand, AD patients who happened to have a stroke would be diagnosed as VD. These results suggest that further investigations as to VD (its clinical course, findings of computed tomographic scan and magnetic resonance imaging) are needed for more appropriate clinical criteria. PMID- 8230790 TI - [Nutritional assessment and rapid turnover protein in the elderly]. AB - Nutritional assessment with respect to rapid turnover protein (RTP) in the elderly was performed in 22 outpatients (7 males and 15 females, mean age 77 years) who suffered from slight illnesses such as mild hypertension and osteoporosis. Their nutritional parameters were almost within the normal range, judging from body mass index (BMI) and their plasma levels of total protein, albumin, lipids and RTP (transferrin, pre-albumin and retinol-binding protein). Statistically, BMI had a significantly positive correlation with subcutaneous fats and total protein and had a significantly negative correlation with HDL cholesterol. Retinol-binding protein had a significantly positive correlation with prealbumin, ferritin and vitamin A. Conversely, in 12 bedridden patients the nutritional status was very poor, with their plasma levels of the above markers being significantly lower than those of the controls. We administered an enteral diet (EWH330) to 10 bedridden patients in an effort to restore their nutritional status. As a result, their nutritional parameters, especially RTP showed a significant increase after 4 and 8 weeks of the administration of EWH330. These results suggest that RTP is a very sensitive parameter of nutritional assessment in the elderly. PMID- 8230791 TI - [Age and gender difference in circadian rhythm in blood pressure]. AB - In order to evaluate the effect of gender and aging on the diurnal rhythm of blood pressure (BP), we performed ambulatory BP monitoring in 267 normotensive volunteers (mean age +/- SD 52.1 +/- 24.6 years, range 16-93 years, 114 men, 153 women). Ambulatory 24-hour BP was recorded every 30 minutes with an oscillometric method by ABPM630 (Nippon-Kolin, Japan). Circadian rhythm was determined by fitting the 24-hour cosine function curve. Subjects were divided into three age groups, consisting of the Y group (age < 30), M group (age > or = 30 but < 60), and E group (age > or = 60). Highest normal values of the 24-hour mean BP (90 percentile of the values) were 126/75 mmHg in the Y group, 126/76 mmHg in the M group, and 154/85 mmHg in the E group in men. They were 117/67 mmHg, 125/77 mmHg 151/79 mmHg respectively in women. The MESOR of SBP increased with age in women, though it was significantly higher only in the E group among men. The amplitude of SBP decreased with age significantly only in men. The acrophase of SBP shifted to an earlier time of day with aging in both men and women. The trends in the circadian rhythm of DBP with age were similar to those of SBP. In conclusion, the circadian rhythm of blood pressure varies with gender and aging. The 24-hour blood pressure values should be considered with evaluating sex-age matched reference values. PMID- 8230792 TI - [Effect of age on cognitive potentials using auditory and colored visual stimulation]. AB - Cognitive potential using two-tone auditory discrimination task has been widely used as a standard and useful test to quantify the severity of cognitive dysfunction. According to our experience, however, this method (the T method), sometimes cannot be performed even in normal elderly subjects because of difficult discrimination between these two different tone bursts. In order to overcome this disadvantage, we developed two other methods which had easier discrimination tasks: one using a tone burst and click sound (the C method), and the other using red and green color (the V method). We adopted these three methods to 64 normal adults and aged subjects whose age ranged from 20 to 88 years old. As a result, most of the P3 latency and amplitude in all subjects tested correlated significantly among these three methods. Any peak latency of P3 correlated significantly (p < 0.05) and positively with age, but the rate of increment of latency per year differed. For example, the value recorded from the Cz scalp site was 0.77 ms/year in the T method, 1.05 ms/year in the C method, and 1.67 ms/year in the V method, respectively. The rate of undetected P3 potential in aged subjects more than 60 years old was higher in the T method (3/18 subjects = 17%) than by both the C and V methods (1/19 subjects = 5%). In conclusion, the effect of age on P3 latency varied according to method and our new technique was sufficiently applicable to elderly subjects. PMID- 8230793 TI - [Effect of gentian violet on the elimination of methicillin-resistant Staphylococcus aureus (MRSA) existing in the decubitus region]. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is frequently isolated from skin lesions, such as in the decubitus region. There is a possibility that MRSA through these lesions can spread widely in a hospital. However, local treatment with most antibiotics and antiseptics (povidone-iodine) is not effective to eradicate MRSA from the infected decubitus. We have recently demonstrated that gentian violet (Gv) possessed a bactericidal effect against MRSA isolated from clinical specimens in vitro. This examination evaluated whether or not a topical ointment containing 0.1% Gv is effective to eradicate MRSA which existed in decubitus regions. Decubitus (14 clinical cases, ages 59-87 years) infected with MRSA were treated with 0.1% Gv-ointment once or twice daily after bathing in 0.1% Gv aqueous solution. Although all patients were treated with povidone-iodine and 9 out of 14 patients were given either local or systemic administration of antibiotics, those treatments were not effective to eradicate MRSA from decubituses. However, MRSA was not detectable in all cases within 34 days (average: 10.8 days +/- 2.7) after treatment with 0.1% Gv-ointment. The eradication of MRSA from decubitus areas tended to be delayed, depending upon the size and depth of decubituses (Grade III and IV) and complications such as diabetes mellitus. Skin irritability was not observed in any patients. These results suggest that 0.1% Gv-ointment is a useful material for the treatment of the MRSA-local wound infection. Treatment with Gv-ointment to MRSA-infected decubitus may exhibit a protective effect with regard to infection with MRSA in hospital. PMID- 8230794 TI - [Elderly cases of hematological malignancies with second malignancies]. AB - Recently an increase of the elderly patients with hematological malignancies has been pointed out. We analyzed second malignancies in elderly patients with hematological malignancies (95 age 65 or more), and made a comparative study with non-elderly case for the past 5 years. Second malignancies were observed in 26 cases out of the total of 282 hematological malignancies (9.2%). The percentage of patients with second malignancies in the elderly group (19/95; 20%) was significantly higher than that of the non-elderly group (7/187; 3.7%). Among the all kinds of hematological malignancies, the second malignancies were mainly observed in cases with myelodysplastic syndrome and chronic myelo-proliferative disorder. Colon carcinoma, gastric carcinoma and lung carcinoma accounted for nearly half of all the second malignancies. On 11 of the 26 cases with second malignancies, the first malignancies had been treated with some anti-cancer drug such as alkylating agents. Development of a second cancer was greater in cases in which the first hematological malignancy was treated with alkylating agents more than in cases in which the first carcinoma was not treated with alkylating agents. PMID- 8230795 TI - [The relative assessment of atherosclerosis in vessels of major organs and its factors for progression]. AB - In order to study the relative assessment of atherosclerosis among arteries of various organ (coronary, aortic, peripheral and cerebral artery), 94 patients aged 43-69 underwent examination by coronary angiography (CAG), pulse wave velocity (PWV), ankle pressure index (API) and funduscopy simultaneously. The severity of coronary sclerosis was determined by the coronary atherosclerotic index (CAI) which was based on the size and number of stenotic lesions, which that of funduscopic findings was evaluated using Scheies' score (S+H). The relationship between risk factors of atherosclerosis and each value for the examinations (CAI, PWV, API, S+H) were studied. Furthermore, subjects were divided into two groups of normal or early change groups and a progressive group based on the degree of atherosclerosis. Then, the difference of atherosclerosis in each vessel of major organs was compared. A positive correlation between PWV and age, and a negative correlation between API and age were noted (p < 0.01, p < 0.05). A higher prevalence of hypertension in groups of progressive PWV or API were observed (p < 0.05). Total cholesterol in the CAI progressive group was significantly higher than in the normal group (p < 0.005), and it became higher in relation to the progression of PWV or API (p < 0.001). HDL-C in the progressive groups of CAI, PWV API and S+H were significantly lower (p < 0.05, p < 0.05, p < 0.02, p < 0.01). On the other hand, glucose intolerance influenced the progression of CAI, PWV, API and S+H, statistically. In subjects whose age was limited to between 55-64, statistically significant correlations were noted between any pair of examinations among CAI, PWV, API and S+H. The percentage of API and S+H progressive groups were lower in CAI normal of PWV early change groups, suggesting earlier sclerotic change of the coronary artery or aorta than that of peripheral or cerebral artery. These results demonstrate that the atherosclerosis in major vessels could be assessed relatively using a non pathological process, which supports the usefulness of these examinations in clinical practice. PMID- 8230796 TI - [The effect on changes in serum lipids and factors that may affect those changes after administration of icosapentaenoic acid ethyl ester]. AB - The effects of high-purity IPA-E administration on changes in serum lipids and factors that may affect those changes were investigated in 55 patients with arteriosclerotic disease (34 males and 21 females, aged 43-85) under a regular daily diet. Daily administration of 1800 mg or 900 mg IPA-E resulted in a significant decrease or tendency to decrease in serum TG, TC and LDL-C 4 weeks after the initial administration, which continued throughout the administration period. Apo-protein analysis revealed a significant increase in A-I and a significant decrease in B and E after the administration of IPA-E. Lipoprotein analysis disclosed decreasing tendency of VLDL, IDL, LDL and TG, where the changes in VLDL-C, VLDL-TG and TG were significant. The amount and rate of change in TG, TC and LDL-C after IPA-E administration had a significant negative correlation with their respective pre-administration values. The decrease from pre-administration values was significant for patients with abnormal values in both dosage groups. The average rate of decrease in TC and LDL-C were greater in the group receiving administration of 1800 mg/day than that receiving 900 mg/day, but the difference in the change was not significant between the 2 groups. Correlations between the amount or rate of decrease in TC or LDL-C and the baseline value, achieved value, or amount or rate of increase in plasma IPA concentration and IPA/AA ratio were not significant. However, the negative correlation between the rate of change in TG and achieved IPA concentration was significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230797 TI - [Clock drawing task as screening for cognitive impairment in the elderly population]. AB - This study compared the "Clock-Drawing Task" to the Mini-Mental State Test (MMST) in 834 elderly persons. A cut-off score of 20 or less on the MMST was used to determine cognitive impairment. The "Clock drawing Task" was score of 20 or less on the MMST was used to determine cognitive impairment. The "Clock drawing Task" was scored by three components; frame, numbers, and two hands. Using a criterion of completion on two or more of the three components, the sensitivity, specificity, and correct identification of the diagnosis of cognitive impairment of "Clock Drawing Task" were 89%, 65% and 83% respectively. "Clock Drawing" appears to be a practical test which can be useful for clinicians as a screening for cognitive impairment in old age. PMID- 8230798 TI - [An elderly patient with acute idiopathic thrombocytopenic purpura (ITP)]. AB - An elderly patient with acute-type ITP is reported. An 89-year-old man was admitted to the Department of Physiatrics, Ureshino-Onsen Hospital on July 19, 1991 and commenced rehabilitation treatment. Laboratory data on January 28, 1992 showed decreased platelets (2.8 x 10(4)/mm3). According to this data, the patient was transferred to the Department of Internal Medicine, where laboratory data showed positive occult blood in urine and stool, prolonged bleeding time (over 10 min.) and positive PAIgG (630 ng/10(7) cells). Bone marrow showed an ITP pattern (megakaryocytes 55/mm3). The patient received steroidal pulse therapy (methylprednisolone 1,500 mg/day for 3 days). Subsequently, serum platelet count increased gradually and reached 10 x 10(5)/mm3 in May 1992. We reviewed the case reports of ITP published in Japanese, and conclude that this case was the oldest reported patient with acute-type ITP in Japan. PMID- 8230799 TI - [An episode of health control on hazardous workers]. PMID- 8230800 TI - [A survey of primary lung cancer among NTT workers in the Tokyo area. The incidence and mortality during the past 30 years and significance of radiologic screening]. AB - An epidemiological study of primary lung cancer was conducted on NTT workers in the Tokyo area (about 40,000 persons). During the past 30 years (1960-1989), 77 cases (71 males and 6 females) of primary lung cancer were detected by routine medical examination (radiologic screening by radiophotography at their workplaces) or by clinical symptoms, of whom 59 (54 males and 5 females) were dead as of the end of December 1991. Standardized incidence rate and standardized death rate of the males by the direct method (5-yr age groups from 20 to 59 yr of age, per 100,000 population) were 4.5 in the 1960's, 9.9 in the 1970's and 9.8 in the 1980's, and 3.3 in the 1960's, 7.9 in the 1970's and 8.2 in the 1980's, respectively, with increase being observed from the 1970's. Standardized incidence ratio and standardized death ratio of the males by the indirect method (5-yr age groups from 20 to 59 yr of age) were 107 in the 1970's, 86.8 in the 1980's, and 53.8 in the 1960's, 99.4 in the 1970's and 98.5 in the 1980's, respectively. No significant difference could be observed when compared to the national average. By histological type, 44 cases of adenocarcinoma, 12 cases of squamous cell carcinoma, 8 cases of small cell carcinoma, 7 cases of large cell carcinoma and 6 cases of other or unclear types were detected. Excluding the 3 unclear cases, of the 77 cases, 54 cases were peripheral type and the remaining 20 cases were hilar type on radiophotography. Peripheral adenocarcinoma was the most common type.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230801 TI - [LD50 and weight change in organs of mice following intraperitoneal administration of methyl tertiary-butyl ether]. PMID- 8230802 TI - [Study on the relationship between appearance of autoantibodies and chest X-ray findings of asbestos plant employees]. AB - Antinuclear factor (ANA) and rheumatoid factor (RF) were determined in 220 employees of an asbestos plant. The relationship of the appearance of the two factors to sex, age, smoking habit, duration of asbestos exposure and asbestos exposure level was studied. In 207 employees who had chest roentgenograms, the relationship of these two factors to pulmonary lesions and pleural lesions was examined. Of the 220 employees, 33 (15%) had ANA and 7 employees (3.2%) had RF. ANA positive rate in the asbestos plant employees was significantly higher than in the control. Factors significantly correlated to their appearance could not be demonstrated. With the progress of pulmonary lesions the ANA positive rate significantly elevated, but ANA positive rate was not significantly correlated with pleural lesions. RF did not show any significant correlation with any of the factors. PMID- 8230803 TI - [Measurement of ethylene oxide at a medical sterilization site]. AB - Although ethylene oxide gas is widely used as a sterilizing agent for medical instruments because of its disinfection property, the effects of its use in medical settings have not been clarified. In the present study, we measured the ethylene oxide gas concentration (EOGC) within a hospital sterilization unit and in the ambient air near the unit. Before the sterilizer was turned on (about 9:00), the ambient air EOGC was below the detection limit (0.1 ppm). When the door was opened to place the instruments in the sterilizer, the maximum EOGC near the door of the sterilizer was 1.71 ppm. Before the sterilizer door was opened, the residual EOGC within the sterilizer was 0.10-24.56 ppm. During the operation of the sterilizer (9:00-17:00), ethylene oxide gas could not be detected in the air near the unit. When the sterilizer door was opened at the end of the routine operation of the sterilizer (about 17:00), EOGC near the door was 2.10-2.73 ppm. After the door was closed, the ambient air EOGC level was 0.5-0.57 ppm. These findings indicate that the personnel near the unit were exposed to ethylene oxide gas for about 15 min during the transfer. However, no ethylene gas could be detected by the ethylene oxide gas monitor (3M Co., #3551). The finding that EOGC in sterilized medical instruments after 24 h of aeration was about 2 ppm also suggests that the personnel using these instruments were exposed to ethylene oxide gas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230804 TI - Biosynthesis of the blood group P antigen-like GalNAc beta 1-->3Gal beta 1- >4GlcNAc/Glc structure: kinetic evidence for the responsibility of N acetylglucosaminyl-transferase. AB - Previously we reported the occurrence of UDP-GalNAc:Gal beta 1-->4GlcNAc/Glc beta 1,3-N-acetylgalactosaminyltransferase activity in human plasma. Here, the donor substrate specificity of the enzyme partially purified from blood group O plasma was investigated by means of competition experiments with analogs of donor. The enzyme activity was found to be inhibited most strongly by UDP-GlcNAc among the nucleotide sugars tested, and UDP was the best inhibitor among test nucleotides. UDP-GlcNAc was much more inhibitory than UDP, whereas UDP-GalNAc was much less inhibitory than UDP. These results show that the donor-binding site of enzyme has a high affinity for UDP-GlcNAc, but not UDP-GalNAc, suggesting that the enzyme essentially functions in the transfer of N-acetylglucosamine. Indeed, UDP GlcNAc:Gal beta 1-->4GlcNAc/Glc beta-1,3-N-acetylglucosaminyltransferase activity is known to occur in human plasma. The ratio of N-acetylgalactosamine to N acetylglucosamine transferred to lactose with O or B plasma was 1:20 when assayed separately at 1 microM of donor, but increased to 1.2:1 at 5 mM. The nearly identical ratio was obtained with the partially purified enzyme preparation. PMID- 8230805 TI - Two different histamine-sensitizing activities of pertussis vaccine observed in mice on the 4th and 12th days of sensitization. AB - The histamine-sensitizing (HS) activities of commercial precipitated diphtheria tetanus-purified pertussis combined vaccines (DTP) submitted to the national quality control tests during the period from 1980 to 1991 were measured in mice on days 4 (EHS) and 12 (LHS) of sensitization. Annual averages of the EHS activity of the vaccines showed continuous decline during the period, while the LHS activity stayed unchanged except for 1990 and 1991. Correlation analysis between these two HS activities revealed that the vaccines could be differentiated into two groups, those showing a significant correlation and those lacking it, depending on their source manufacturers. When the vaccine was incubated at 37 C for four weeks, both the HS activities increased for the first couple of weeks at different rates, reaching their peaks after different periods; three weeks for the EHS activity and two weeks for the LHS activity. Treating the reference pertussis vaccine (for toxicity tests) with anti-pertussis toxin horse serum neutralized completely the EHS activity but the LHS activity resisted the serum. These findings suggest a possible difference between the two HS activities in their mode of action, therefore, in their roles in possible reactogenicity of the vaccine, and a necessity for separate controls of the two activities. PMID- 8230806 TI - Epidemiology of hydatidosis and echinococcosis in Theqar Province, southern Iraq. AB - Ninety-six cases of human hydatidosis caused by Echinococcus granulosus were diagnosed from surgical records of hospitals in Theqar Province, Southern Iraq during 1989. The infection rate was higher in female than in male patients (56 versus 40 cases) and increased with their age (maximum, 25.0% in a female group of 31-40 years and 32.5% in a male group of 41-50 years). Many organs such as liver, lungs, peritoneum and spleen were involved, but the liver was most frequently affected. Of 96 patients, 66 (68.8%) showed single organ-involvement and 30 (31.2%) multiple organ-involvement. In 50 stray dogs surveyed for E. granulosus the prevalence was found to be 56.0% with a mean worm burden of 5,300. The light infection (1-200 worms) was found in 10.7% of the infected dogs, the medium (201-1,000 worms) in 21.4% and the heavy (over 1,000 worms) in 67.9%. PMID- 8230807 TI - In vitro gametocytocidal activity of artemisinin and its derivatives on Plasmodium falciparum. AB - Artemisinin and six of its derivatives were evaluated for their in vitro gametocytocidal and erythrocytic schizontocidal properties on an Indian isolate of Plasmodium falciparum. One of the metabolic derivatives, DADF dihydroartemisinin (NIH02), was found to possess gametocytocidal and erythrocytic schizontocidal properties similar to those of artemisinin. Gametocytes of this isolate were highly susceptible to the toxic effect of NIH02 (IC50 = 6.6 ng/ml) and younger stages were more sensitive. This is the first report about the in vitro gametocytocidal properties of a derivative of artemisinin. PMID- 8230808 TI - Identity of pathogenic strains of spotted fever rickettsiae isolated in Shikoku District based on reactivities to monoclonal antibodies. AB - Three IgG (B2, C3 and F8) and two IgM (S3 and X1) monoclonal antibodies (Mab) were produced in BALB/c mice immunized with Aoki strain of spotted fever (SF) rickettsiae in Tokushima Prefecture, Japan. Of these, two (B2 and C3) reacted equally with six Japanese strains including YH strain, the prototype of R. japonica, but did not react with the foreign strains. These results indicate that SF rickettsial strains from Tokushima Prefecture were identified as R. japonica, and that the SF strains isolated at least in Shikoku District most likely belong to a common serotype. PMID- 8230809 TI - Immunological comparison of intracellular toxin A and extracellular toxin A from Clostridium difficile. AB - Intracellular toxin A and extracellular toxin A of Clostridium difficile were compared immunologically. Immunodiffusion tests with anti-intracellular toxin A and anti-extracellular toxin A sera showed that these toxins were identical. In neutralization tests, both antisera neutralized the homologous and heterologous toxins with regard to cytotoxicity, mouse lethality and loop response at nearly the same titers. Although intracellular toxin A lacks the hemagglutination (HA) activity, anti-intracellular toxin A serum neutralized HA activity of extracellular toxin A at the same titer as did anti-extracellular toxin A serum. These results suggest that these toxins are immunologically indistinguishable and that the intracellular toxin A molecule has an antigenic site(s) for the HA activity. PMID- 8230810 TI - [Changes in anionic sites on glomerular basement membrane following mesangiolysis -observation in incipient anti-rat Thy1 nephritis]. AB - To clarify whether heparan sulfate proteoglycans (HSPG) on glomerular basement membrane (GBM) are produced by mesangial cell in vivo, the number of anionic sites (AS) on lamina rara externa of GBM and the changes in urinary albumin excretion rate (UAE) were observed on mesangiolysis in rats induced by monoclonal anti-rat Thy1 antibody. Mesangiolysis was remarkable at day 3 after the administration of anti-rat Thy1 antibody, whereas GBM as well as epithelial cell remained well preserved, and detached endothelial cell was observed. The number of AS on GBM was significantly (p < 0.001) decreased in anti-rat Thy1 antibody treated rats (18.6 +/- 0.4/1000nm), when compared with that of normal control rats (22.8 +/- 0.4/1000nm). Furthermore, UAE was markedly (p < 0.05) increased at day 3 (1692.6 +/- 627.5 micrograms/24h) compared with the value before (188.2 +/- 30.6 micrograms/24h) administration of anti-rat Thy1 antibody. From these results, it can be concluded that some part of AS on GBM may be produced by the mesangial cell. PMID- 8230811 TI - [Immunohistochemical localization of epidermal growth factor in proximal tubules of rats with acute renal failure]. AB - Increasing evidence supports the hypothesis that epidermal growth factor (EGF) may have a role in the repair of tubular injury on acute renal failure (ARF). On the other hand, platelet-depletion reduces the renal damage in experimental ARF rats. We investigated the expression of EGF in the renal tissues of mercuric chloride (HgCl2, 2mg/kg)-induced ARF rats with (group-1) or without (group-2) platelet depletion for clarification of the role of EGF in the recovery of the renal tubules after ARF. Platelet-depletion was accomplished by the injection of anti-rat platelet serum (APS) 24 hours prior to the induction of ARF. The tubular necrosis and deterioration of renal function were significantly suppressed in group-1 compared with group-2 24 hours after HgCl2 injection (S-Cr 0.60 +/- 0.17 mg/dl, Ccr 0.58 +/- 0.11 ml/min in group-1 and 1.45 +/- 0.10 mg/dl, 0.20 +/- 0.02 ml/min in group-2, p < 0.05) but were in the same range between both groups 12 hours after (S-Cr 0.75 +/- 0.03 mg/dl, Ccr 0.49 +/- 0.02 ml/min in group-1 and 0.60 +/- 0.05 mg/dl, 0.67 +/- 0.08 ml/min in group-2). By immunohistochemistry, EGF was localized in the ascending limbs of Henle and distal convoluted tubules in normal rats. However, in both ARF rats, EGF was stained on the proximal tubules, including the necrotic area as well as the ascending limbs of Henle and distal convoluted tubules, both 12 and 24 hours after ARF induction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230812 TI - [Systemic hemodynamics and renal blood flow in glycerol induced acute renal failure]. AB - In our previous papers, we reported change in the systemic hemodynamics in rats with glycerol-induced acute renal failure, as estimated by the microsphere method. It was found that the development of acute renal failure related not only to renal blood flow, but also to the hepatic blood flow. In view of the liver playing an important role in the metabolism of substances, such as drugs, and in blood circulation, we examined changes in the systemic circulation and blood chemistry in rats with thioacetamide-induced liver cirrhotic, and those rats injected with glycerol. The result indicated that liver cirrhotic rats showed hypertension, higher levels of atrial natriuretic peptide (ANP) and lower levels of renal blood flow relative to the control rats. However, after glycerol injection, the development of acute renal failure was suppressed in liver cirrhotic rats. These results suggest that liver cirrhotic rats were in a hyperdynamic state and that this condition would be beneficial in the prevention of glycerol-induced development of acute renal failure. PMID- 8230813 TI - [Electron microscopy study on alterations of glomerular basement membrane in IgA nephropathy]. AB - To elucidate the morphological basis of glomerular hematuria in IgA nephropathy, morphometric analysis of GBM alterations was performed by electron microscopy in 73 cases of IgA nephropathy. These cases were divided into 2 groups by the degree of hematuria. The IgA (1) groups (52 cases) showed 1-30 Red Blood Cells (RBC)/hpf in urinary sediment and the IgA (2) group (21 cases) showed 30 or more RBC/hpf. The GBM alterations were observed in 67% of IgA nephropathy patients. Splitting and focal thinning of GBM were more frequent in the IgA (2) group than in the IgA (1) group. The gap was not associated with the degree of hematuria, but when accompanying splitting, it was with the degree of hematuria. It was concluded that most IgA nephropathy patients had GBM alterations, and splitting of the lamina densa, focal thinning of GBM, and a gap with splitting, which was associated with the degree of hematuria. PMID- 8230814 TI - [Severe proteinuria in IgA nephropathy. Clinicopathological study of 8 cases with 100 serial sections]. AB - Using 100 serial sections for light microscopy, we studied the clinicopathologic characteristics of IgA nephropathy with severe proteinuria. Eight out of 128 cases with IgA nephropathy exhibited severe proteinuria of 3.0g/day or more. These cases consisted of 5 males and 3 females with an age range from 15 to 73 years. Daily proteinuria ranged from 3.3 to 7.1 g. At the time of biopsy, the serum creatinine level was 1.2 mg/dl or more in 6 cases; two of these developed end-stage renal failure during the follow-up period. In most of the cases, routine sections for light microscopy revealed moderate to severe mesangial proliferation and tubulointerstitial damage. In 7 cases, variable percentages of glomeruli, ranging from 20% to 95%, were involved by chronic lesion represented by sclerosis and fibrous crescents/adhesion, while only 2 cases exhibited active lesion characterized by segmental glomerular necrosis and/or cellular/fibrocellular crescents. In contrast, thorough observation of 100 additional serial sections disclosed the focal presence of active lesion in all except one case with features of endstage kidney. In addition, the highest percentage of glomeruli with active lesion in the serial sections correlated well with the severity of proteinuria. The results suggested that severe proteinuria in IgA nephropathy depends on, at least in part, focal outbreak of active lesion characterized by segmental glomerular necrosis and cellular/fibrocellular crescents. PMID- 8230815 TI - [Clinical and pathological study of lupus nephritis without clinical renal involvement]. AB - To analyze the clinical and pathological changes in systemic lupus erythematosus (SLE) patients without clinical renal abnormalities, I performed histopathological examinations of renal biopsy specimens from 40 SLE patients, who were divided into two groups. Group A consisting of 16 patients had almost no clinical evidence of renal involvement. Group B consisting of 24 patients had clinical renal manifestations, proteinuria, hematuria or both due to lupus nephritis. Light microscopy showed minor to moderate proliferative changes in the patients of Group A, and severer changes in the patients of Group B. Immunofluorescent study revealed depositions of immunoglobulins and complement in varying degrees in all the patients of Groups A and B. By electron microscopic study, electron-dense deposits and mesangial deposits were found in all patients of Group A. However, subendothelial, subepithelial and intramembranous deposits were seen in less than one-half of the patients of Group A. The amounts of the deposits were small. By contrast, a large amount of electron-dense deposits existed in the patients of Group B. The prognosis was better for patients of Group A than those of Group B. I concluded that various abnormalities of the renal glomeruli exist in SLE patients even without urinary abnormalities. PMID- 8230816 TI - [The clinical significance of albuminuria in renal transplantation]. AB - Urinary excretion of albumin was measured after renal transplantation in 37 patients who received cyclosporine quadrotherapy (cyclosporine A, azathioprine, prednisolone and antilymphocytoglobulin). In 26 patients with a favorable clinical course, daily urinary albumin level increased significantly on the first operative day. Subsequently, it decreased gradually, and remained at about 40mg/day after 3 weeks. In these patients, the urinary albumin level did not correlate with blood pressure, whole blood cyclosporine level, serum creatinine level nor urinary enzyme level, i.e. beta 2-microglobulin, NAG, gamma-GTP. As for correlation between the urinary albumin level and biopsy findings in 21 patients who underwent renal graft biopsy, the urinary albumin level was significantly increased in 5 patients with high degree of lymphocyte infiltration in the renal graft as compared with 16 patients with low degree of infiltration. However, there was no significant correlation between the urinary albumin level and the degree of tubular damage. In 4 patients of allograft rejection, a clinical course was reflected better in the albumin level than in the urinary enzyme level, these results show that the measurement of urinary albumin is a useful clinical examination to evaluate the allograft rejection of renal transplantation in cyclosporine era. PMID- 8230817 TI - [Accumulation of a disopyramide metabolite in renal failure]. AB - Twenty-nine cases of hypoglycemia induced by disopyramide (DP) have been reported in the literature to date. Twenty of the reported cases showed hypo-renal function and a high concentration was rare. DP is metabolized to mono-N dealkyldisopyramide (MND) in the liver and accumulation of MND is to be expected in renal failure. Both DP and MND bind mainly to alpha-1-acid glycoprotein (AAG) in the plasma. In 10 hemodialysis (HD) patients with normal liver function receiving DP therapy in the steady state. DP, MND and AAG were measured pre- and post-HD. Ten patients with normal renal and liver function were selected as the controls. The DP concentration was 2.08 +/- 0.39 micrograms/ml (mean +/- SD) in the control group, and the pre- and post-HD levels were 2.40 +/- 1.08 micrograms/ml and 1.73 +/- 0.87 micrograms/ml, respectively, in the HD group. The MND concentration was 0.42 +/- 0.23 micrograms/ml in the controls, 1.53 +/- 0.52 micrograms/ml in pre-HD and 1.08 +/- 0.32 micrograms/ml in post-HD. Although DP and MND are both classified as substances of small molecular weight, the average decrease in plasma concentration from pre- to post-HD was under 30% with both agents. The MND/DP ratio in the HD group was higher than in the controls, but there was no significant difference between pre- and post-HD. The AAG level was 75 +/- 5mg/dl in the controls and 109 +/- 11mg/dl before HD in the HD group (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230818 TI - [Transcatheter arterial embolization for severely dysfunctioning kidney]. AB - The authors performed Transcatheter Arterial Embolization (TAE) with pure ethanol in five cases of severely dysfunctioning kidneys, for which conventional surgical nephrectomy had been conducted. In 3 out of 5 cases, the dysfunctioning kidneys were complicated by renal hypertension, while the other two had the complication of refractory urinary fistula. 1. Single TAE trials in 4 out of 5 patients, and a secondary TAE trial in the remaining patient resulted in successful complete embolization of the kidneys. 2. The size of the kidney in the bidirectional measurement by CT after TAE was reduced markedly in all the five cases and the reduction rate of the kidney was from 89.29 to 62.84%, and 72.61% on the average. 3. Both the blood pressure (BP) and plasma renin activity (PRA) in the three patients with complicated renal hypertension were all normalized two weeks after the TAE. Twenty six months, 48 and 60 months have elapsed in these three cases and blood pressure has remained normal without antihypertensive agents. From this result it is considered that TAE will induce the reduction of juxtaglomerular cell functions. 4. Two patients with urinary fistula were healed. 5. TAE has a number of advantages over surgical nephrectomy in that it is bloodless and noninvasive. Further, the techniques of TAE are easier than nephrectomy and TAE is applicable to almost all poor risk patients. PMID- 8230819 TI - [Maintenance therapy on the anemia in continuous ambulatory peritoneal dialysis (CAPD) patients using subcutaneous administration of recombinant human erythropoietin fortnightly--a multicenter trial]. AB - Eighty-seven CAPD patients whose hematocrit (Ht) level was maintained by recombinant human erythropoietin (rHuEPO) were enrolled in this trial for a new formulation of rHuEPO suitable for subcutaneous injection. 6000IU rHuEPO was administered every 2 weeks for 12 weeks. Fortnightly doses were increased to 9000IU or 12000IU at 4 or 6 weeks if the Ht level decreased by 2% or more. During the study period, Ht values were maintained at the appropriate level in 88% of patients. 6000IU or lower was selected as a maintenance dose given every 2 weeks in 57 (76.0%) patients, 9000IU was selected in 8 patients and 12000IU was chosen in one patient. In 9 patients, the Ht could not be maintained during the study and the appropriate dose, therefore, remained unclear. Hypertension was observed in 2 patients as a side effect, and headache occurred in 2 other patients during the trial. Cutaneous abnormalities were not observed in the course of the subcutaneous injections. We conclude that anemia in most CAPD patients could be controlled with fortnightly injections in the dose range of 6000 to 12000IU rHuEPO given subcutaneously. PMID- 8230820 TI - [A case of non-IgA mesangioproliferative glomerulonephritis with huge paramesangial hemispherical deposits]. AB - A 16-year-old female was admitted to our hospital because of chance proteinuria. On admission, mild proteinuria (0.6g/day) was observed, but microhematuria was not detected during the observation period. All the values of the renal function tests were within the normal range. Her renal biopsy demonstrated a prominent increase in the mesangial area by light microscopy and showed marked paramesangial hemispherical deposits by electron microscopy. Though IgM, IgG, Clq, C3, and fibrinogen were localized in the mesangial region, IgA was not detected by immunofluorescence study. It has been reported that paramesangial hemispherical deposits are strongly indicative of IgA glomerulonephritis. We conclude that this patient is a rare case of non-IgA glomerulonephritis with huge paramesangial hemispherical deposits. PMID- 8230821 TI - [Nephrotic syndrome with microcellular projections into the thickened basement membrane]. AB - A 47 year-old patient with an 8-year history of proteinuria was admitted to our hospital in 1989. His laboratory data were compatible with nephrotic syndrome: total serum protein 5.9g/dl (albumin 3.0g/dl), total serum cholesterol 280mg/dl and total urinary protein excretion 5.0g/day. Renal biopsy specimens contained 27 glomeruli associated with sclerosis and collapse of loops under light microscopical examination. In addition, mesangial proliferation and thickening of the basement membrane were visualized. With immunofluorescent study granular IgG deposits were detected in the peripheral region of the glomeruli. Staining for IgA, IgM, C3, Clq, light chain and Congo-red were all negative. On electron microscopy, microtubules apparently resembling cellular projections appeared to thicken the basement membrane. We are tempted to conclude that the current case is an atypical glomerulopathy accompanied by a glomerular microtubular-like structure. PMID- 8230822 TI - [A case of acute hepatitis A associated with acute renal failure from the onset]. AB - A 42-year-old female was admitted to a hospital, because of acute hepatitis A. Laboratory data were GOT 8210mU/ml. GPT 4650mU/ml, LDH 11860mU/ml, total bilirubin 4.7mg/dl, BUN 19.5mg/dl and creatinine 1.9mg/dl. Urinalysis showed proteinuria 3+ and occult blood 1+. Soon after admission, she suffered from anuric acute renal failure and was transferred to our hospital for hemodialysis. Her urine-volume was under 20 ml per day. Urinalysis showed proteinuria 4+, occult blood 1+ and casts. Laboratory data showed BUN 58.2mg/dl and creatinine 8.5mg/dl. She was treated by hemodialysis for 35 days, before recovering from renal failure. However, her renal function did not recover perfectly and her 24 hour creatinine clearance remained at 50ml/min after 6 months. Renal biopsy was performed on the 17th day after admission. Examination by light microscopy revealed the findings of acute tubular necrosis and examination by immunofluorescence antibody method was negative. Urinalysis of 8 patients with acute hepatitis A showed that all patients had proteinuria at the onset. Patients with acute hepatitis A have symptoms of appetite-loss, nausea, vomiting and/or diarrhea. These symptoms cause hypovolemia, and hepatic dysfunction causes discontrol of vasoactive hormones, which gives rise to disturbance of renal circulation. Subsequently, acute tubular necrosis and acute renal failure occur. PMID- 8230823 TI - [A long-term hemodialysis patient complicated with systemic calciphylaxis]. AB - A 46-year-old male patient underwent long-term hemodialysis treatment had suffered from calciphylaxis (defined by Selye), such symptoms as advanced systemic vascular calcification, rapid progression of gangrene on both fingers and toes, disturbance of consciousness, and sclerosis and obstruction of the superficial vein after venipuncture during 11.5 years of dialysis. Furthermore, he had a long history (30 years) of heavy smoking. He died as a result of sepsis due to pneumonia after 12.5 years of dialysis. He had received dialysis treatment using a small amount of dialysate (50 liters on a recirculating system) for 8.5 years and had been dialysed 2 and 2 or 3 times a week for 10 years. As a result of this insufficient dialysis treatment, his characteristic laboratory data showed hypocalcemia, hyperphosphatemia, elevated calcium-phosphorus product, advanced metabolic acidosis, hyperalkaliphosphatemia and elevated serum parathyroid hormone. Autopsy revealed the following: 1) enlargement parathyroid gland enlarged in two (4.0 g and 2.0 g, respectively) showing adenomatous hyperplasia presenting cord-like arrangement of chief cells and water-clear cells, 2) systemic medial calcification in radial, ulnar, renal, mesenteric and brain arteries, and 3) Berline-blue positive iron deposit in calcified arteries in mesenteric and parathyroid tissue. From these results, we concluded that factors (challengers) related to the appearance of calciphylaxis might be as follows: 1) advanced secondary hyperparathyroidism, 2) long-term uremic state, 3) administration of VD2 and VD3, 4) iron salt injection, and 5) a long history of heavy smoking. We speculated that these challengers might act synergistically to cause calciphylaxis. PMID- 8230824 TI - [Relationship between reduced myocardial uptake of beta-methyl-p-(123I)- iodophenyl-pentadecanoic acid (123I-BMIPP) and regional diastolic ventricular dysfunction in patients with hypertrophic cardiomyopathy]. AB - To assess whether regionally depressed myocardial uptake of beta-methyl-p-(123I) iodophenyl-pentadecanoic acid (123I-BMIPP), is related to regional ventricular diastolic dysfunction in patients (pts) with hypertrophic cardiomyopathy (HCM), we carried out 123I-BMIPP myocardial SPECT and radionuclide ventriculography (RNV) at rest in 9 HCM pts (eight pts with asymmetric septal hypertrophy (ASH), and one patient with apical hypertrophy). The defects of 123I-BMIPP were semiquantitatively estimated by segmental analysis. Regional LV function was determined by sector analysis in RNV: regional peak filling rate (rPFR), regional time to peak filling rate (rTPFR), and early 1/3 filling rate (1/3 fil) were estimated in the LV septal and posterior sectors. Wall thickness was evaluated with M-mode echocardiography. As results, the defects of 123I-BMIPP myocardial SPECT, were found frequently in septal wall, but any defect was not recognized in all septum with significant hypertrophy. Also, the degree of septal hypertrophy was not significantly related to BMIPP defect score. However, the regional diastolic function (rPFR, rTPFR, 1/3 fil) in the septal lesions with 123I-BMIPP defects was decreased more than those without 123I-BMIPP defect in HCM. Furthermore, the relative regional diastolic dysfunction (the differences between rTPFR and 1/3 fil in the septum and in the posterior wall) was significantly related to BMIPP defect score. In conclusion, this results suggest that the decreased uptake of 123I-BMIPP is related to regional diastolic dysfunction more than wall hypertrophy. PMID- 8230825 TI - [Survey for primary tumor site in patients with initial clinical presentation of bone metastasis]. AB - Among the patients who were examined with bone scintigraphy between April 1985 and March 1991, there were 27 patients whose initial clinical manifestation was bone metastasis and who were surveyed for the primary tumor site. The primary tumor site could be identified in 20 patients (74%), consisting of 9 patients with lung cancer, 3 with prostate cancer, 3 with hepatoma, 2 with renal cancer, and one each with thyroid cancer, adrenal cancer, and pleural malignant mesothelioma. In 17 of the 20 patients, the primary site had been detected within two months after presentation. Examinations which were helpful in identifying the primary site included chest radiography, sputum cytology, abdominal sonography, serum prostatic acid phosphatase level and pathologic examination of biopsy specimens. 99mTc-PMT scintigraphy was useful in the diagnosis of the hepatoma when accumulation was observed at the metastatic sites. In 2 patients, lung cancer had been recognized using follow-up chest radiography 3 and 6 months after presentation, respectively. One patient was diagnosed at autopsy as having adrenal cancer. In 7 patients the primary site remains unknown. Histology examination of the biopsy specimen performed in 6 of these patients revealed 4 to be adenocarcinoma and 2 undifferentiated carcinoma. The average survival period of the 17 patients who died was 9.5 months. Four patients are alive, and the outcome in the remaining 6 could not be determined. PMID- 8230826 TI - [Estimation of effective half life of 131I in radioiodine therapy for Graves' disease using a single radioiodine uptake measurement]. AB - Calculation of the absorbed radioiodine dose in the thyroid gland is a very important factor in 131I therapy for Graves' disease. In 131I therapy, the measurement of the effective half life (EHL) of radioiodine in the thyroid gland is indispensable for calculating the absorbed radioiodine dose. In some cases, however, EHL cannot be calculated because repeated measurements of thyroid uptake cannot be done. So, the authors developed a new method to estimate EHL using a single radioiodine uptake measurement. The activity of intrathyroidal 131I (thyroid uptake) and that of the surrounding area of the shielded thyroid (body background) were measured at 24 hours after the administration of 131I. The ratio of the value of body background to thyroid uptake was calculated. This ratio was multiplied by 100 and the calculated value was named the EHL-index. The relationship between actual measured EHL and the EHL-index was expressed by a regression line and/or a hyperbola. For the purpose of obtaining more actual values of EHL using the EHL-index, the new combined curve was composed of the regression line and the hyperbola. As for the value of calculated EHL compared with that of actual measured EHL, about 80 percent of calculated EHL values were within acceptable error ranges. These results indicate that EHL can be estimated by a single measurement 24 hours after administration of 131I. This method should be very useful in the therapy of Graves' disease with 131I when the thyroid measurement is performed only one time. PMID- 8230827 TI - [The utility of three-phase bone scintigraphy in the assessment of fractured carpal scaphoid]. AB - Thirty-seven joints in 36 patients with a fractured carpal scaphoid were evaluated by three-phase bone scintigraphy. They were classified into two groups according to their progress. Some were in good clinical condition and some in a non-union condition. Increased blood flow in the radial arteries and ample perfusion on and around the scaphoid bones on blood flow images suggested a good clinical course. The activity and the effectiveness of remodeling correlated well with the degree of scaphoid uptake on blood pool images taken more than seven days after the injury. Scaphoid uptake was more localized or there was almost none on blood pool images in cases with nearly complete recovery while it was amply visualized on static images. Blood pool images were indispensable for analyzing lesions and evaluating the clinical course. Two typical findings of scaphoid fractures were found on both blood pool and static images. One was diffusely increased scaphoid uptake seen in cases with a good clinical course, and the other was decreased uptake at proximal fragments seen in cases with non union. It is concluded that three-phase bone scintigraphy provides useful information for evaluating the process of scaphoid fractures which cannot be obtained by means of conventional bone scintigraphy. PMID- 8230828 TI - [Acetazolamide effect on vascular response in crossed cerebellar diaschisis as measured by 99mTc-HMPAO SPECT]. AB - We studied the effect of acetazolamine (DIAMOXR) on vascular response in areas with crossed cerebellar diaschisis (CCD) using consecutive 99mTc-hexamethyl propyleneamine oxime (HMPAO) SPECT studies before and after DIAMOX administration with a subtraction method. Regions of interest were drawn over the bilateral cerebellar cortices and interhemispheric asymmetry indices (IAI) were obtained. Eleven of 14 patients with CCD at baseline showed decrease of IAI after DIAMOX administration. Although there is a possibility of underestimation of increase in perfusion in unaffected cerebellar hemisphere, this result suggests a greater increase in perfusion in the affected cerebellar hemisphere as compared with in the contralateral unaffected one and suggests more dilatation of the arterioles in areas with CCD post-DIAMOX than in areas without CCD. However nonsignificant correlation between IAI at baseline and difference of IAI from baseline to post DIAMOX did not support the hypothesis of more dilatation post-DIAMOX of more constricted arterioles. PMID- 8230829 TI - [Sequential changes in the left ventricular function during ramp exercise in patients with ischemic heart disease: continuous assessment of the left ventricular radioactivity with wireless telemetric cardiac monitoring system]. AB - The left ventricular function during ramp exercise was assessed using 99mTc labeled red blood cells and a Wireless Telemetric Cardiac Monitoring System (WTCMS) in 34 patients who received coronary angiography. Sequential changes in the left ventricular ejection fraction (EF) during exercise, were classified into the two types, normal response (up slope pattern) and abnormal response (horizontal, down slope and up-down slope pattern) when defined the response as 5 percent or more increment of EF as normal. Six patients without significant stenosis by coronary angiography showed normal response. Seventeen of 28 patients with coronary disease showed abnormal response (sensitivity 61%). Eight of 11 patients (73%) with coronary disease and normal response showed no redistribution on the stress thallium myocardial scintigraphy. Twelve of 17 patients (71%) with coronary disease and abnormal response showed redistribution by the stress thallium scintigraphy. Horizontal and down slope patterns of EF during exercise were more often noted in the groups with redistribution and multivessel coronary disease. After the exercise, the times to the over shoot was significantly longer in groups with redistribution (p < 0.01) in comparison with groups without redistribution. From these results, we conclude that the monitoring the changes in left ventricular activity during ramp exercise with the WTCMS is a very useful method for the evaluation of the sequential changes in left ventricular function in patients with ischemic heart disease. PMID- 8230830 TI - [A case of radiation-induced chronic constrictive pericarditis developing 16 years after irradiation]. AB - We reported a 51-yr-old female with radiation-induced chronic constrictive pericarditis. At age 29, she had received a mastectomy and postoperative irradiation because of left breast cancer. At age 45, she had syncope and was diagnosed with complete atrioventricular block and a pacemaker was implanted. At that time, pericardial thickening with effusion was noted. The following year, tricuspid regurgitation was noted. On catheter study, a dip and plateau pattern of the right ventricular pressure curve appeared. At age 50, tricuspid regurgitation worsened due to the lead wire of the pacemaker compressing the leaflet, and the pacemaker was reimplanted. However, the following year, she complained of general fatigue and dyspnea and was admitted to our hospital. On 67Ga study, diffuse accumulation in the cardiac region appeared. There was no perfusion defect detected in the left myocardium, but right myocardial damage was suspected by thallium study. In 99mTc-HSA RI angiography, right atrium dilatation appeared and a pericardial halo around the ventricles was seen. She underwent pericardectomy, tricuspid replacement and pacemaker reimplanted, but she died. On autopsy, pericardial thickening and adhesion, right myocardial fibrosis, the fibrotic change of the bundle branches were seen. We reported a case of radiation induced constrictive pericarditis. Radionuclide studies were useful in diagnosing and following the patient. PMID- 8230831 TI - [A case of benign paraganglioma arising in the middle mediastinum; 201Tl-SPECT for differentiation from malignant tumor]. AB - A case of benign paraganglioma arising in the middle mediastinum was reported. 201Tl SPECT showed high accumulation in tumor on early images at 15 min and reduced on late images at 3 hours after infusion. The patient was a 57 year old female. In the contrast enhanced CT, 3 x 4 cm tumor with intensive enhancement was recognized at the right middle mediastinum. Under the radiological images, the tumor was surgically removed. The pathologic diagnosis was a low atypical nonfunctioning aortico-pulmonary paraganglioma. This report was suggested that 201Tl SPECT was useful for differential diagnosis of benign neurogenic mediastinal neoplasms. PMID- 8230832 TI - [Usefulness of serum pancreatic phospholipase A2 determination in patients with various pancreatic diseases]. AB - A radioimmunoassay kit (SHIONORIA P-PLA2) to measure human serum pancreatic phospholipase A2 (P-PLA2) concentrations was evaluated for their basic properties and clinical usefulness. The performance of the kit was found to be quite satisfactory. To test its clinical usefulness, specimens mainly from patients with pancreatic diseases were obtained and examined for serum P-PLA2 levels. All serum specimens from patients with acute pancreatitis (n = 7) were found to have elevated levels of serum P-PLA2. In patients with chronic pancreatitis (n = 93) and pancreatic cancer (n = 37), serum P-PLA2 levels showed a wide range of distribution, from abnormally elevated values to abnormally low values. We compared these data with those of other pancreatic markers such as: amylase, elastase 1, trypsin, and PSTI. Among these, P-PLA2 was highly specific to pancreatic disease and best represented the state of pancreatic disorders. Abnormally elevated levels of serum P-PLA2 concentrations seem to reflect the inflammation of pancreas, while abnormally low levels indicate the hypofunction of pancreatic exocrine glands. These results suggest that the measurement of serum P-PLA2 concentrations is a useful diagnostic test for pancreatic disorders. PMID- 8230833 TI - [A basic study of multi-gated single photon emission computed tomography with thallium-201]. AB - A Multi-gated SPECT was acquired commonly in 64 x 64 matrix and 8 frames per cardiac cycle (64 x 64/8F). But it was not established that 64 x 64 matrix and 8 frames per cardiac cycle were the most suitable in multi-gated SPECT. Five normal volunteers were examined multi-gated 201Tl SPECT with 5 acquisition modes of 128 x 128 matrix/16 frames, 128 x 128/8F, 64 x 64/32F, 64 x 64/8F using multi detector SPECT system (GCA-9300). And we calculated percent wall thickening (%WT) [%WT = (ES counts - ED counts)/ED counts] in 9 cases with 64 x 64/8F. The images quality of both 128 x 128/16F and 128 x 128/8F was not clear in compared with images of both 64 x 64/16F and 64 x 64/8F, because the end-diastolic phase of 128 x 128/16F images showed a decreased uptake of 201Tl in the antero-apical region. Although 64 x 64 (8) images had only 8 frames per cardiac cycle, we could observe systolic and diastolic phase and we could calculate %WT. The %WT (M +/- SD) of horizontal long axis images were 48 +/- 15 (sept. basal), 48 +/- 19 (sept. apical), 65 +/- 29 (apex), 49 +/- 22 (lat. apical) and 40 +/- 15 (lat. basal).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230834 TI - Immunohistochemical localization of epidermal growth factor in human main and accessory lacrimal glands. AB - Immunohistochemical localization of epidermal growth factor (EGF) in human main and accessory lacrimal glands has been investigated using monoclonal and polyclonal antibodies. Positive stain of EGF was localized in the cytoplasm of the acinar cells and the epithelial cells of the intra- and interlobular ducts of the main lacrimal glands, including both the palpebral and orbital lobes, and of the accessory lacrimal glands (gland of Krause, gland of Wolfring) as well. Meibomian glands showed weak immunoreactivity. Positive stain in the conjunctival epithelium including goblet cells was not observed. These results suggest that the mature form of EGF is produced in both the main and accessory human lacrimal glands and secreted into the tear fluid as a constant and crucial component. PMID- 8230835 TI - Critical flicker frequency in acute and recovered optic neuritis. AB - Multiple occasional changes in critical flicker frequency (CFF) were studied in patients who recovered from optic neuritis. Twenty-five patients (31 affected eyes) with onset visual acuity less than 0.5 and who showed recovered visual acuity exceeding 1.0 were included in this study. Recovery stages were determined individually as follows: T1, initial onset stage; T2, intermediate stage; T3, recovered stage when visual acuity was 1.0 or better; and T4, final follow-up stage. CFF was determined using red, yellow and green illuminated targets in a compact CFF measuring device recently developed at our department. The rates of abnormality were 100% at stage T1 for all colors, and gradually decreased as the stage advanced from T2 to T4. However, the rates of abnormality continued to be high at 67% in stage T3 for the red target and 37% in stage T4 for the red target. The rates of abnormal interocular difference in CFF in 15 unilateral optic neuritis patients were 100% for all colors at stage T1 and decreased gradually with recovery. Slight but definite abnormality of CFF was also noted in the silent eyes of clinically unilateral optic neuritis patients. The rates of abnormal CFF more than 7% in all colors could be detected in T1, T3 and T4. These results indicate that CFF abnormality can be detected even at the stage of recovery in the pathologic eyes and the fellow eyes of optic neuritis patients. CFF was also shown to be a sensitive indicator for detecting visual dysfunction in patients with optic neuritis. PMID- 8230836 TI - Photopic electroretinogram implicit time in retinitis pigmentosa. AB - While the implicit time of cone responses in the electroretinogram (ERG) has been suggested to have some clinical implications in retinitis pigmentosa (RP), its evaluation is not always easy in cases of typical RP because of the small amplitudes of the responses. An intense flash stimulus would be expected to improve the recordability of photopic ERG, which represents the cone responses. However, the implicit time in normal eyes has been shown to be delayed with increased stimulus intensity. Therefore, we evaluated photopic ERGs to a white flash stimulus of 3 different intensities as well as to a red flash in 14 cases of typical RP. The recordability of the response to white flash improved with the increased intensity of the stimulus. Most RP cases showed prolonged implicit time compared with that in normal eyes to each flash stimulus. Some patients who showed exceptionally normal or mildly delayed implicit time were considered to have good prognosis since they had noticed night blindness at an older age and most of their rod response in ERG was recordable. PMID- 8230837 TI - Clinical study of venous abnormalities in diabetic retinopathy. AB - The authors studied 304 eyes (248 cases) affected by diabetic retinopathy in relation to venous abnormalities. The posterior pole and surrounding mid peripheral area were examined by ophthalmoscopy, slit-lamp examination with contact lens and fluorescein angiography. There were 225 eyes with preproliferative and 79 eyes with proliferative retinopathy. Fifty-six eyes (18%) of the total eyes had venous abnormalities: 17 (8%) of the 225 eyes with preproliferative retinopathy and 39 (49%) of the 79 eyes with proliferative retinopathy. In the 56 eyes with venous abnormalities, 54 (96%) showed beading, 14 (25%) showed looping and 2 (4%) showed duplication. All eyes which showed looping or duplication had proliferative retinopathy. The locations of the venous abnormalities were analyzed: 42% were in secondary branches, 27% in tertiary branches, 18% in primary branches, 10% in fourth branches and 3% in fifth branches. The portion of the fundus affected was upper temporal, lower temporal, upper nasal and lower nasal in 28%, 22%, 29% and 21%, respectively. Based on our results, eyes with looping or duplication on ophthalmoscopic or slit-lamp examination have an extremely high probability of being in some stage of proliferative retinopathy. Retinopathy should be confirmed by fluorescein angiography and photocoagulation should be performed as quickly as possible. PMID- 8230838 TI - A Japanese family with Grayson-Wilbrandt variant of Reis-Bucklers' corneal dystrophy. AB - A 50-year-old man (the proband) experienced recurring attacks of ocular irritation and had had bilateral ring-shaped anterior corneal opacity since adolescence. Corneal sensation was normal. The proband's 15-year-old daughter also had recurring episodes of ocular irritation and bilateral fleck-like anterior corneal opacities. The proband's deceased father and brother reportedly had similar corneal lesions from adolescence. We believe that the corneal findings in this family may be similar to those found in patients with the Grayson-Wilbrandt variant of Reis-Bucklers' corneal dystrophy. PMID- 8230839 TI - Long-spacing collagen in the human corneal stroma. AB - An electron microscopic observation of a corneal lenticule removed 14 months after epikeratoplasty because of refractive error and an unused lenticule was carried out to determine the presence of long-spacing collagen. In the removed lenticule, long-spacing collagen, which is often described as the product of the keratocytes migrating from the host corneal stroma, was observed near a keratocyte. However, long-spacing collagen was also observed in the unused lenticule. In this specimen, keratocytes had been destroyed by freezing so that the secretion of long-spacing collagen by keratocytes should have been negligible. Thus, it seemed that long-spacing collagen could exist naturally in the corneal lenticule as well as be newly formed by migrating keratocytes. In addition, we examined four corneas from patients ranging from 2 months to 91 years of age with no past history of corneal diseases or disorders. Long-spacing collagen was seen in the corneal stromas of the aged persons, but not in those of the younger. In the corneas of the 78-year- and 91-year-old, a different appearance from the usual long-spacing collagen was also seen in the collagen fibrils which showed a slightly shorter periodicity resembling experimentally formed 100 nm periodic fibrils in mice. Long-spacing collagen is a common component of normal human corneal stroma and its occurrence seems to correlate with the age-related changes of the tissue. PMID- 8230840 TI - A multicenter study of typical retinitis pigmentosa in Japan. AB - A nationwide, multicenter study of typical retinitis pigmentosa was carried out in collaboration with 13 university hospitals throughout Japan. A total of 253 patients, 122 males and 131 females, with a wide range of ages (mean 48 years), were registered during a two-month period in 1989. Determination of inheritance pattern revealed 30.2% autosomal recessive cases, 15.4% autosomal dominant, 0.5% X-linked, and 48.9% simplex, indicating a relative decrease in autosomal recessive cases and a relative increase in simplex cases in recent decades. The age at onset, initial symptom, and visual functions including visual acuity, visual field and electroretinogram showed a marked interindividual variability, but statistical analysis demonstrated that visual defects progressed with increasing age and disease duration. A correlation between the phenotypic variation and the genetic type was observed. This survey of retinitis pigmentosa in Japan provides information for counseling and rehabilitation of patients and encourages basic and clinical research of this genetic disease. PMID- 8230841 TI - Ophthalmic diseases in bedridden patients with severe dementia. AB - Ophthalmologic examinations were performed on the 176 eyes of 88 bedridden patients with severe dementia. There were some pathologic changes in the external and anterior segments of 93 eyes. We observed infectious external and anterior segment eye diseases in 80 of these 93 eyes (86.0%). Fundus diseases were found in 34 eyes, but treatment was not necessary. Infectious eye diseases appeared to be the most prevalent ocular problem in these patients. Cultures of conjunctival swabs commonly demonstrated Staphylococcus aureus. No differences were noted in tear secretion rate or lysozyme concentrations in tears between eyes with and without infection. The blinking frequency was significantly reduced in infected eyes compared to normal eyes. Failure of the lacrimal drainage system was observed more often in infected eyes than in normal eyes. This study demonstrated that bedridden patients with severe dementia develop infectious eye diseases easily, to which stasis of tear flow seems to predispose. In caring for bedridden patients with severe dementia, ophthalmologists must be concerned with controlling infectious eye diseases. PMID- 8230842 TI - Saucerisation (recession) of neuro-retinal rim is characteristic of glaucoma. AB - Colour stereophotographs of the optic disc and surrounding retina were assessed by a Humphrey analyser system in a series of Japanese individuals. When compared with 9 age- and sex-matched control eyes (9 controls), the 10 eyes of 10 patients with early open-angle glaucoma showed a significantly lower level of the mid point of the neuroretinal rim relative to the level of the surface of the retina about 1/4 of a disc diameter beyond the disc edge. When the mid-point of the rim was related to the surface of tissue at the disc edge, the level of the former was significantly lower at the three measurement points on the nasal side and at 6 o'clock. In the 9 normal controls, the lower half of the neuro-retinal rim was at a significantly lower level than the upper half, relative to the surface of tissue at the disc edge. We conclude that saucerisation and/or recession of the neuro-retinal rim frequently accompanies glaucoma and we suspect it is an early sign of it. PMID- 8230843 TI - An application of laser speckle phenomenon for noninvasive two-dimensional evaluation of microcirculation in ocular fundus--a preliminary report. AB - A new instrument has been developed which uses the laser speckle phenomenon for noninvasive two-dimensional analysis of the microcirculation in the optic nerve head (ONH). The ONH is illuminated by a diode laser spot and its image speckle is detected by an area sensor. The difference between a pair of output data from successive scannings of the image speckles at the sensor plane is calculated and integrated for each pixel to obtain the average difference (AD) value, which is a quantitative index of blood flow velocity. The results are displayed in color graphics showing the two-dimensional variation of the AD value in the area of interest. In an in vitro model, the AD value correlated well with the speed of rotation of a ground glass in a range between 0 and 5 mm/second. Using this instrument, the effect of intraocular pressure (IOP) on the circulation was studied in the area of the rabbit ONH free of surface vessels. The coefficient of reproducibility was 4.7% when measured twice in this area at 5-minute intervals. A stepwise reduction in the ocular perfusion pressure (OPP) was introduced by elevating the IOP manometrically. The AD value was little affected by a change in OPP when the OPP was above 40 mmHg; while when the OPP was below 40 mmHg, a linear correlation was found between the AD value and the OPP. This finding agreed well with those of previous studies where the effect of the OPP on the ONH blood flow were studied using various invasive methods.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230844 TI - A case-control study of risk factors for proliferative vitreoretinopathy in aphakia. AB - A case-control study was performed to elucidate the risk factors of proliferative vitreoretinopathy (PVR) in aphakia. Twenty-five aphakic eyes with rhegmatogenous retinal detachment accompanied by PVR were compared with the control group of 156 aphakic eyes with rhegmatogenous retinal detachment without PVR. Unconditional logistic regression analysis identified the following risk factors for developing PVR in order of significance: choroidal detachment (odds ratio 15.41, 95% confidence interval 3.29-72.09), duration of retinal detachment longer than one month (odds ratio 12.07, 95% confidence interval 3.53-41.26), occurrence of retinal detachment within one year following cataract surgery (odds ratio 6.2, 95% confidence interval 1.64-23.47), and history of vitreous loss in cataract surgery (odds ratio 3.91, 95% confidence interval 1.28-11.92). PMID- 8230845 TI - Inhibition of in utero galactose-induced cataract development by an aldose reductase inhibitor in rat--electron microscopic study. AB - The effect of an aldose reductase inhibitor (ARI) on the development of in utero galactose-induced cataract was evaluated by following the morphological changes as seen by light and transmission electron microscope. Pregnant rats were fed a 30% galactose diet until day 17, 18, 19 or 20 of gestation and then given a galactose diet containing ARI. The lenses of newborn rats when mothers were given an ARI diet after day 17, 18 or 19 of gestation showed no morphological change. On the other hand, in the lenses from newborn rats when mothers were given an ARI diet after day 20 of gestation, 30% of those tested showed small vacuoles limited to the posterior cortex, while 70% showed normal structure. When mothers were given a galactose diet during the entire term of pregnancy all lenses of newborn rats showed extensive vacuole formation. ARI was confirmed to completely inhibit congenital galactose cataracts in rats when it was given to the mother rat after day 19 of gestation (ie, on the last 3 days before delivery). PMID- 8230846 TI - Vitreous hemorrhage associated with acute posterior vitreous detachment--a case report. AB - A case of vitreous hemorrhage associated with acute posterior vitreous detachment was reported. The patient was a 64-year-old man who complained of floaters in his left eye of one-week duration. His previous medical and ocular histories were unremarkable. Slit-lamp examination with Goldmann three-mirror contact lens showed complete posterior vitreous detachment in both eyes. Ophthalmoscopic examination and fluorescein angiogram revealed a partial defect in retinal blood flow in the midperipheral area of the superotemporal region of the left fundus. On follow-up examination, recovery of the retinal circulation was observed in that region and the vitreous hemorrhage had spontaneously resolved. The diagnosis was made in the left eye of rupture of retinal vein due to vitreous traction associated with acute posterior vitreous detachment without retinal break formation. PMID- 8230847 TI - Diagnostic evaluation of diabetic retinopathy by 30-Hz flicker electroretinography. AB - The peak latency in 30-Hz flicker electroretinography (ERG) was determined in patients with diabetic retinopathy, using a newly developed contact lens electrode with built-in light-emitting diode. A good correlation was obtained between the peak latency of flicker ERG and that of oscillatory potentials of flash ERG. The peak latency of the flicker ERG was only slightly prolonged in background retinopathy, but markedly prolonged in proliferative retinopathy. The stage of pre-proliferative retinopathy with normal peak latency in flicker ERG may be a compensatory state of the retina, which is the most appropriate stage for laser treatment of diabetic retinopathy. PMID- 8230848 TI - Pulsatile sweating and sympathetic sudomotor activity. PMID- 8230849 TI - A novel H+,K(+)-ATPase in the colonic apical membrane? PMID- 8230850 TI - Electrical responses of the rat's retinal pigment epithelium to azide and thiocyanate. AB - In the rat under urethane anesthesia, a fast intravenous injection of a bolus of sodium azide elicited a transient cornea-positive change in transocular potential (azide response). A bolus injection of sodium thiocyanate (NaSCN) produced a cornea-negative response (SCN- response) with a similar rising phase as the azide response, but with a faster return from the peak. The peak amplitude depended on bolus volume, concentration, animal strain, and age. For more than 24 h, the azide and SCN- responses could be recorded repeatedly from a single rat with little variation in peak amplitudes. Following an administration of iodate, known to degenerate the retinal pigment epithelium (RPE), the transocular d.c. potential decreased; the azide response became smaller and then was inverted in polarity, whereas the SCN- response became larger. Azide and SCN- are assumed to depolarize and hyperpolarize the basal membrane of RPE, respectively. The equilibrium potential of ions passing through the putative azide-sensitive channels is assumed less negative than resting potential of RPE cells. The SCN- response probably represents a diffusion potential of SCN- permeating through anionic channels at a higher rate than Cl-. Results demonstrate the feasibility of in vivo electrophysiological measurement of the functional state and the structural integrity of RPE under pathological conditions. PMID- 8230851 TI - In vivo response of the rat's retinal pigment epithelium to azide: changes induced by light damage. AB - Functional changes in retinal pigment epithelium (RPE) associated with light induced retinal damage were studied by measuring transocular potential changes evoked by injections of azide and thiocyanate (SCN-). The retinal damage by light in the rat is classified into two types: Type 1, rod cell death associated with RPE deterioration; Type 2, the loss of rod cells without RPE deterioration. To study the type 1 damage, littermate pairs of long-term dark-adapted adult albino rats were tested at 1 h and 10 d after the exposure to green light of 1,200 lx for 1/2 to 24 h. Time course of the damage progress was also followed for 12 h. We found that 1) RPE was affected rapidly by the damaging light, 2) the exposure length determined the ultimate degree of RPE damage, 3) damaging effects on RPE proceeded slower and weaker after exposure than during continuous light, 4) progress of the damage in RPE was two-phasic; during the first phase, the SCN- response was enhanced and the azide response was reduced; both responses were decreased rapidly in the second phase. The first phase was assumed to indicate a depolarization of the basolateral membrane of RPE, and the second phase to manifest the structural deterioration of RPE. The type 2 damage was studied in young rats with exposure to weak light for 28 d. At 30 d after the exposure, a wave of the ERG and number of rod cells were substantially reduced but azide and SCN- responses were affected slightly. PMID- 8230852 TI - In vivo response of the rat's retinal pigment epithelium to azide at advanced stages of hereditary retinal dystrophy. AB - Electrophysiological properties of the retinal pigment epithelium (RPE) were studied in the rat with hereditary retinal dystrophy (rdy). Transocular potential changes evoked by intravenous bolus injections of azide and thiocyanate (SCN-) are the only available indication of RPE state when degeneration of rods is in progress. Also determined were age-dependent decrease in retinal DNA content and in counts of cones that survive after degeneration of rods. The azide response in the pigmented and albino rdy rat was already reduced at the earliest age tested (60 d) and continued to decrease till the age of 2 years. The SCN-response was similarly affected but seemed to decline faster than the azide response. The azide/SCN- response ratio was significantly increased in albino mutants, especially around the age of 400 d. At the age of 10 months and later, the azide and SCN- responses became slower than those of normals. A prolonged exposure of 1,200 1x light to dystrophic rats older than 110 did not affect the azide and SCN responses whereas the same exposure abolishes the responses of normal rats and of the dystrophic rats at early stages. In rdy rats, the electrophysiological changes were considered to correlate with structural changes of the junctional RPE complex and with abnormal membrane enzyme distribution discovered by others. These RPE changes may contribute to the decreasing cone cell number after rod cell disappearance. PMID- 8230853 TI - Cardiovascular responses to gastric hypo-osmolar stimulation in anesthetized dogs. AB - We compared the cardiovascular response to the gastric infusion of distilled water (DI) with that to the gastric infusion of 0.9% saline (SI) and gastric ballooning with 37 degrees C water (BA) through a gastric fistula in splenectomized mongrel dogs (n = 7). DI, and SI amounting to 5% of body wt and the same volume of water were infused in approximately 20 s through the tube and responses in mean arterial pressure (MAP), CVP, heart rate (HR), and intra esophageal pressure (EP) were monitored continuously. After DI, SI, and BA, the measured variables showed significant increases and attained maximal increases at about 2 min after the treatments. After DI, the maximum elevation in MAP was 21.3 +/- 1.9 mmHg and 2 times higher than in SI and BA. The corresponding value in CVP was 5.0 +/- 0.3 mmHg and 2-3 times higher than with SI and BA, and HR increased by 26.1 +/- 3.0 beats/min showing 3 to 6 times larger increases compared with SI and BA. These gastro-cardiovascular reflexes were abolished after subdiaphragmatic truncal vagotomy. These findings suggest that both the mechanical and osmotic stimuli to the stomach induce cardiovascular reflexes and that the vagus is involved in the reflex. PMID- 8230854 TI - Maximal oxygen uptake and aerobic-anaerobic transition on treadmill and bicycle in triathletes. AB - The maximal aerobic capacity and the aerobic-anaerobic transition were analyzed on 14 triathletes performing an incremental work load on a bicycle ergometer and on horizontal or inclined treadmills. To compared the cardiorespiratory responses between cycling and running, the subjects were divided into 2 groups of 7 with similar aerobic capacity determined from cycle runs. The first group ran on horizontal treadmill while the second group performed similar exercise on inclined treadmill at constant grade (1.5%). Heart rate was recorded by electrocardiogram. Oxygen uptake (VO2), CO2 production (VCO2), respiratory frequency, and pulmonary ventilation were monitored at 30 s intervals through a Rudolph valve connected to a calibrated Oxycon V. Tidal volume, respiratory exchange ratio, equivalent O2 and CO2 were calculated from on-line computer. Aerobic and anaerobic thresholds were determined by a non-invasive method from pulmonary ventilation curves. The results showed that maximum oxygen uptake (VO2max) did not differ between the 2 types of ergometers. Pulmonary ventilation, heart rate and VO2 recorded at aerobic and anaerobic thresholds depended on the mode of exercise and reached the highest values on inclined treadmill. The amount of muscle mass, the type and the distribution of active motor units involved in each exercise test might be at the origin of these differences. This indicates that, when assessing a training program from anaerobic threshold values, it is necessary to take into consideration the type of ergometer used and the protocol performed. PMID- 8230855 TI - Augmented arterial to end-tidal PCO2 difference during laparoscopic CO2 insufflation in man. AB - Acid-base status of the blood and tissue fluids and pulmonary gas exchange were continuously observed during intra-abdominal CO2 insufflation for 2 h in 8 paralyzed and artificially ventilated patients who underwent laparoscopic cholecystectomy or resection of the sigmoid colon. Carbon dioxide pressure in the arterial and mixed venous blood as well as in end-tidal air started to increase already at 5 min CO2 insufflation. On the other hand, appreciable elevation in VCO2 and tissue PCO2 (PTCO2) was noted only after 15 min. In vivo buffer value (beta) was well within the normal physiological range at 5 min and gradually declined up to 2 h. These observations indicated that chemical buffering to CO2 and redistribution of buffer base among the blood and tissue fluids were slowly developed during entire period of observation. In reflecting these experimental findings, arterial to end-tidal PCO2 difference (a-ADCO2) and respiratory gas exchange ratio (R) were promptly and significantly increased at 5 min and maintained slow increment up to 2 h. We conclude that these profiles of a-ADCO2 and R can be explained by initially rapid and subsequently slow augmentation in ventilation-perfusion ratio (VA/Q) during the specific type of respiratory acidosis elicited in this study. PMID- 8230856 TI - Hepatic extraction and hepatic action of insulin, glucagon, and epinephrine in bivascularly perfused rat liver. AB - Using a bivascularly perfused rat liver, we investigated the hepatic extraction and hepatic action on glucose output of insulin, glucagon, and epinephrine. The liver was perfused for 70 min without recirculation via the portal vein and hepatic artery (3.0 ml/(min.g liver) from portal vein and 1.0 ml/(min.g liver) from celiac artery). Low and high concentration of insulin, glucagon, or epinephrine (450 and 4,500 pM, 21.5 and 215 pM, or 410 and 4,100 pM, respectively) was added to the perfusing medium via portal vein (P-liver), via celiac artery (H-liver), or via portal vein and celiac artery (H-P-liver). Hepatic extraction of insulin, glucagon, or epinephrine was not significantly different among H- (56 +/- 9 and 21 +/- 5%, 31 +/- 7 and 24 +/- 6%, or 22 +/- 8 and 17 +/- 5%), P- (50 +/- 10 and 23 +/- 6%, 32 +/- 9 and 23 +/- 7%, or 22 +/- 9 and 18 +/- 6%), and H-P-liver (55 +/- 11 and 24 +/- 6%, 30 +/- 9 and 26 +/- 8%, or 20 +/- 9 and 17 +/- 6%). Glucagon- or epinephrine-induced increase in glucose output was also similar in H- (245 +/- 47 and 498 +/- 82 mumol/30 min, or 112 +/- 21 and 215 +/- 38 mumol/30 min), P- (258 +/- 51 and 512 +/- 95 mumol/30 min, or 128 +/- 20 and 220 +/- 36 mumol/30 min), and H-P-liver (263 +/- 58 and 515 +/- 94 mumol/30 min, or 129 +/- 22 and 225 +/- 39 mumol/30 min).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230857 TI - Left ventricular ORS widening decreases Emax without lowering VO2-PVA relation in dog hearts. AB - We observed a few rare spontaneous cases of a suddenly widened QRS wave of left ventricular ECG associated with a simultaneous decrease in left ventricular (LV) contractility (Emax, end-systolic pressure-volume ratio) in excised cross circulated dog heart experiments. The decreased Emax was not associated with a descent of the relation between cardiac oxygen consumption (VO2) and LV systolic pressure-volume area (PVA, a measure of total ventricular mechanical energy). This result is intriguing because ventricular VO2-PVA relation generally changes its elevation in proportion to Emax under various inotropic interventions. We suspected the unusual observation to reflect no change in myocardial contractility despite ventricular asynchrony augmented by an intraventricular conduction defect. PMID- 8230858 TI - Regional vasoconstriction and excessive grooming induced by L-arginine injection into the cisterna magna of conscious rats. AB - Hemodynamic and behavioral changes caused by intracisternally injected L-arginine which has a pressor effect were studied in conscious rats. One week or more after inserting a cannula into the cisterna magna, electromagnetic flow probes were implanted around the superior mesenteric artery, renal artery, or terminal aorta under anesthesia. A catheter for measurement of blood pressure was inserted into the terminal aorta via a femoral artery or into the common carotid. After recovery from surgery, blood flow, arterial blood pressure, heart rate, and behaviors were observed in the conscious state. Peripheral resistance, calculated as pressure divided by flow, increased an average of 80% in the superior mesenteric artery and 60% in the renal artery when the pressure was increased by 25% following intracisternal injection of 5 mumol of L-arginine. Hindquarter (terminal aortic) resistance showed an insignificant decrease of about 10%. The average heart rate showed no significant change. Grooming, moving, and eating all increased significantly after injection of L-arginine. Circulatory events and behaviors did not always synchronize after the injection. Intravenous chlorisondamine chloride, a ganglion blocker, significantly attenuated the pressor effect caused by intracisternal injection of L-arginine. These findings indicate that centrally injected L-arginine not only specifically triggers vasoconstriction in the viscera presumably through excitation of the regional sympathetic vasoconstrictor fibers to raise blood pressure but also centrally induces excessive grooming. However, the relationship between circulation and behavior caused by intracisternal injection of L-arginine remains unknown. PMID- 8230859 TI - Intracellular Na+ activity of "diseased" human atrial muscles and its modifications by dihydro-ouabain. AB - In quiescent human atrial muscles obtained from patients with various cardiac diseases, the intracellular Na+ activity (aNai) and the resting membrane potential (Vm) were recorded simultaneously, using double-barreled Na(+) selective microelectrodes. The Vm averaged -44.1 +/- 5.0 mV and the aNai, 6.9 +/- 1.7 mM under normal Tyrode's solution containing 5.4 mM [K]oi,The aiNa value was within the range of aNai reported in various cardiac tissues from intact animals. Dihydro-ouabain (10(-5) M) significantly increased the aNai and depolarized the Vm. The results suggest that even in these depolarized atrial muscles the aNai remains within physiologic levels and that the Na-K pump may not be impaired. This is the first report to measure the aNai in human atrial muscles. PMID- 8230860 TI - Human aortic smooth muscle cells containing angiotensin II type 1 receptors. AB - We characterized the angiotensin II (AII) receptor in human aortic smooth muscle cells (HASMCs). This receptor binds [125I]Sar1,Ile8-angiotensin II with a high affinity of 0.20 +/- 0.04 nM and a low capacity of 5.3 +/- 0.4 fmol/mg protein (230 +/- 17 sites/cell). Based on the Ki values, the ranking order of [125I]Sar1,Ile8-AII binding inhibition was as follows: Sar1,Ile8-AII > AII > Dup 753 > AII > AI >> PD 123319. The addition of AII to HASMCs induced a rapid, transient increase in intracellular free Ca2+ concentration followed by a lower, sustained phase. When extracellular Ca2+ was removed by adding 3 mM EGTA, this initial transient increase was not changed, but the sustained phase was abolished. These results revealed AII receptors in HASMCs to be of the type 1 receptor subtype, which induce Ca2+ mobilization mainly from intracellular Ca2+ stores. PMID- 8230861 TI - Involvement of beta-adrenergic systems in the antagonizing effect of paeoniflorin on the scopolamine-induced deficit in radial maze performance in rats. AB - Paeoniflorin, a major constituent of peony root, has been demonstrated to attenuate the radial maze performance deficit produced by scopolamine. In the present study, to investigate the possible involvement of beta-adrenergic systems in the paeoniflorin antagonism of the scopolamine deficit, the effects of two beta-adrenoceptor antagonists, propranolol and atenolol, on the paeoniflorin effect were examined in male Wistar rats. Paeoniflorin (1 mg/kg, p.o.) significantly attenuated the scopolamine HBr (0.3 mg/kg, i.p.)-induced deficit in the choice accuracy in radial maze performance without changing the running time prolonged by scopolamine. Neither D,L-propranolol HCl, a lipophilic beta antagonist, at 3 mg/kg, i.p. nor atenolol, a hydrophilic beta 1-antagonist that is known to hardly ever cross the blood-brain barrier, at 1 mg/kg, i.p. impaired maze performance by itself or aggravated the scopolamine-induced deficit in radial maze performance. Both antagonists, however, completely blocked the antagonizing effect of paeoniflorin on the scopolamine deficit. These data suggest that the beta-adrenergic systems, especially peripheral beta 1-adrenergic systems, are involved in the antagonizing effect of paeoniflorin on the scopolamine deficit in radial maze performance in rats. PMID- 8230862 TI - Important role of peptide leukotrienes (p-LTs) in the resting tonus of isolated human bronchi. AB - The quality of the resting tonus in isolated human bronchi was investigated using a peptide leukotriene (p-LT) antagonist, a 5-lipoxygenase inhibitor and others. (E)-2,2-Diethyl-3'-[2-[2-(4-isopropyl)-thiazoyl]ethenyl]succina nilic acid sodium salt (MCI-826), a newly synthesized compound that is a highly selective antagonist to LTD4 and LTE4, markedly relaxed the isolated human bronchi at low concentrations. A selective and competitive arachidonate 5-lipoxygenase inhibitor, 2,3,5-trimethyl-6-(12-hydroxy-5,10-dodecadiynyl)-1,4-benzoquinone (AA 861), also potently lowered the tonus. In addition, a large amount of spontaneously formed p-LTs was detected in the isolated human bronchial tissue as well as the lung parenchymal tissue. The isolated human bronchi responded to indomethacin treatment with contractions and the acceleration of p-LT formation. Atropine, an anticholinergic; mepyramine, an antihistaminic; and OKY-046, a thromboxane synthetase inhibitor, all showed no effect on the resting tonus. Taking into consideration the high responsiveness of the human airway smooth muscle to p-LTs and the present results, which were different from those on isolated guinea pig tracheas, it is strongly suggested that the spontaneously formed p-LTs largely participate in the resting tonus of the majority of isolated human bronchi. PMID- 8230863 TI - Biochemical and pharmacological properties of a newly synthesized proton pump (H+/K(+)-ATPase) inhibitor, TY-11345 in experimental animals. AB - We investigated the effects of the newly synthesized proton pump inhibitor TY 11345, (+/-)-2-[(4-methoxy-6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin- 9 yl)sulfinyl]-1H-benzimidazole sodium salt, on gastric mucosal proton pump (H+/K(+)-ATPase) activity, gastric acid secretion and gastro-duodenal lesions in experimental animals. TY-11345 potently inhibited H+/K(+)-ATPase activity in isolated rabbit gastric mucosal microsomes; and the inhibitory effect was enhanced under weak acid conditions, the IC50 (concentrations that inhibit the enzyme activity by 50%) being 5.8 microM and 9.9 microM at pH 6.0 and pH 7.4, respectively. In Ghosh & Schild rats, intravenous injection of TY-11345 significantly inhibited gastric acid secretion stimulated by tetragastrin; the effect of TY-11345 was twice as potent as that of omeprazole. In pylorus ligated rats, TY-11345 inhibited basal gastric acid secretion by both the intraduodenal and oral routes, with ED50 values of 1.2 and 4.0 mg/kg, respectively. These effects were 9 and 5 times more potent than those of omeprazole, respectively. Moreover, the antisecretory effect of TY-11345 persisted for more than 24 hr in pylorus ligated rats. In experimental ulcer models, TY-11345 prevented the formation of water-immersion stress, ethanol or indomethacin-induced gastric lesions and mepirizole-induced duodenal lesions in rats. The antiulcer effects of TY-11345 were 3 to 15 times more potent than those of omeprazole. These results suggest that TY-11345 has potent antisecretory and antiulcer effects which are exerted by suppression of H+/K(+)-ATPase activity in gastric parietal cells, so that TY-11345 should be useful for the clinical treatment of peptic ulcer diseases. PMID- 8230864 TI - Chronotoxicity and chronopharmacokinetics of methotrexate in mice: modification by feeding schedule. AB - The circadian rhythms of the toxicity and the pharmacokinetics of methotrexate (MTX), as well as the effects of manipulation of feeding schedule on the rhythms, were investigated in mice. Male ICR mice were housed under a standardized light dark cycle (12:12) with food and water ad libitum (ALF) or under the time restricted feeding (TRF) schedule (8 hr during the light phase) for 1 day or 14 days before the drug administration. The animals received MTX (100 mg/kg, i.p.) once daily for 7 days in the toxicity studies and a single dose of MTX (100 mg/kg, i.p.) for the kinetic studies. Under the ALF, a significant dosing time dependency was demonstrated for the toxicity of MTX with a longer survival time for the middark dosing and a shorter one for the midlight dosing. The MTX kinetics also showed a significant rhythm, with the highest clearance at middark and the lowest one at midlight. The rhythm in MTX kinetics well coincided with that in the toxicity of the drug. The TRF had a marked influence on the rhythms of MTX kinetics and toxicity. Thus, the timing of dosing is important in the kinetics and the toxicity of MTX in mice, and the manipulation of feeding schedule can modify the rhythm of the toxicity by changing that of the MTX kinetics. PMID- 8230865 TI - Mechanism of isoprenaline-stimulated diacylglycerol formation in rat parotid acinar cells. AB - The kinetics and mechanism of sn-1,2-diacylglycerol (DAG) formation induced by isoprenaline were studied in rat parotid acinar cells. DAG accumulation induced by 100 microM isoprenaline reached its maximum at 1 min, rapidly decreased (about 50%) at 5 min and then remained constant for 30 min. DAG accumulation 1 min after isoprenaline treatment was dose-dependent. Either propranolol or phentolamine inhibited isoprenaline-stimulated DAG accumulation in a dose-dependent manner. Addition of a vasoactive intestinal polypeptide, forskolin, or dibutyryl cyclic AMP had no effect on DAG accumulation. Isoprenaline did not cause the release of [3H]choline or [3H]ethanolamine metabolites into the medium. Based on the kinetics of DAG formation and [32P]phosphoinositide breakdown, we conclude that isoprenaline-induced DAG formation was mainly related to the hydrolysis of [32P]phosphatidylinositol 4,5-bisphosphate ([32P]PIP2). These results suggest that the effect of isoprenaline on DAG formation is mediated by alpha 1 adrenoceptor activation, that it is not related to the increase in cyclic AMP, and that it is closely related to PIP2 hydrolysis. PMID- 8230866 TI - Dissociation of cyclic GMP level from relaxation of the distal, but not the proximal colon of rats. AB - The role of cyclic GMP (cGMP) in nonadrenergic, noncholinergic (NANC) relaxation of the longitudinal muscle of rat proximal and distal colon was examined. Electrical field stimulation (EFS) of preparations of longitudinal muscle from the proximal region significantly increased the cGMP content. Nitro-L-arginine inhibited this increase, and L-arginine reversed the inhibitory effect of nitro-L arginine. Exogenously added nitric oxide (NO) and atrial natriuretic peptide (ANP) also increased the cGMP content of preparations of the proximal colon and induced muscle relaxation. From these and our previous findings suggesting an essential role of NO in NANC inhibition in the proximal colon, we conclude that the mechanism of NANC inhibition in the proximal region of rat colon involves NO and a cGMP generating system. In contrast, although exogenously added NO and ANP increased the cGMP content in the distal colon to the same extent as in the proximal colon, they did not induce any muscle relaxation. Vasoactive intestinal peptide (VIP), the most likely candidate as a NANC neurotransmitter in rat distal colon, did not increase the cGMP content in this region. Furthermore, no participation of NO in the NANC inhibitory response was observed in the distal region, but EFS increased the cGMP content significantly. Thus we conclude that relaxation of longitudinal smooth muscle in the distal portion of rat colon is not associated with a change in the cGMP content. PMID- 8230867 TI - Stimulatory effects of HSR-803 on ileal motor activity. AB - Stimulatory effects of HSR-803 on intestinal motor activity in vitro were studied in guinea pig ileum. HSR-803 (1 x 10(-6)-1 x 10(-4) M) increased the amplitude of longitudinal muscle contractions and increased the frequency of peristalsis in isolated segments of guinea pig ileum. The stimulatory effect in amplitude and not frequency was abolished by 1 x 10(-6) M atropine. In the Magnus method with ileal segments, HSR-803 (1 x 10(-7) - 1 x 10(-4) M) produced contractions concentration-dependently, which were inhibited by atropine (1 x 10(-8) and 3 x 10(-8) M) and 3 x 10(-7) M tetrodotoxin (TTX). In the [3H]-quinuclidinyl benzilate (QNB) binding experiment with ileal smooth muscle, HSR-803 had low affinity for acetylcholine (ACh) receptors (pKi = 4.47 +/- 0.04). In addition, HSR-803 failed to increase the spontaneous release and the electrical stimulation induced [3H]ACh release in ileal smooth muscle. On the other hand, HSR-803 (1 x 10(-5) M) enhanced contractions induced by ACh, but had no effect on contractions induced by carbachol, which is not hydrolyzed by acetylcholinesterase (AChE). In conclusion, HSR-803 stimulated ileal motor activity. However, HSR-803 had low affinity for ACh receptors and had no influence on ACh release. It is likely that HSR-803 stimulated motor activity mainly due to prevention of ACh hydrolysis. PMID- 8230868 TI - Effect of systemic anaphylaxis on the hepatic drug-metabolizing system in rats. AB - Rats were immunized by intraperitoneal injection of ovalbumin emulsified with Freund incomplete adjuvant, and then the effect of an intravenous challenge with ovalbumin on hepatic drug-metabolizing enzyme activities was examined. The cytochrome P-450 content and ethylmorphine N-demethylase, benzphetamine N demethylase, arylhydrocarbon hydroxylase and 7-ethoxycoumarin O-deethylase activities significantly decreased in rats treated with ovalbumin compared with control groups treated with saline, whereas there was no significant reduction in cytochrome b5, NADPH-cytochrome c reductase and NADH-cytochrome c reductase. PMID- 8230869 TI - Measurement of oxygen-derived free radical generation in the regionally-ischemic rat heart by the chemiluminescence method. AB - Generation of oxygen-derived free radicals (oxy-radicals) in the stored rat heart was measured by chemiluminescence. Hearts subjected (ischemia) or not subjected (non-ischemia) to a 4-min regional ischemia were frozen in liquid nitrogen and stored until assayed. The frozen myocardium was ground and oxygenated by mixing it with phosphate-buffered saline (Po2: 194 mmHg) containing lucigenin. The chemiluminescence intensity of ischemic myocardium was larger than that of non ischemic myocardium. Recombinant human superoxide dismutase significantly decreased these intensities. These results indicate that O2- is one of the major oxy-radical species in the rat heart and that the generation of oxyradicals is enhanced by regional ischemia for 4 min. PMID- 8230870 TI - [Personality, hassles, and psychological and physical well-being]. AB - The effects of personality characteristics on psychological/physical well-being of ninety-one Japanese working women were investigated with second-order factors of the 16PF questionnaire. Multiple regression analyses revealed significant, negative contributions of extroversion and independence to depression. However, adding hassles as perceived stressors to the regression equation markedly decreased the amount of depression variance explained by these personality variables. None of the personality factors predicted physical symptoms, and only hassles significantly contributed to their prediction, indicating strong associations between hassles and physical symptoms. Conceptual overlap and confounding effects between hassles and symptom measures are discussed in the light of these results. PMID- 8230871 TI - [Analysis of left unilateral spatial neglect with a non-visual sensory-motor task]. AB - An experiment was conducted to analyze the space-movement processes which unilateral spatial neglect (USN) patients manifested. Three kinds of movement features were tested which were reported in previous studies; (a) general coarse response tendency, (b) hypokinesia in the disordered space-side, and (c) more frequent response toward the normal space-side. Five left USN patients with right brain damage were compared with five non-USN right brain damaged patients, and five normal controls. As an experimental movement task, a non-visual movement reproduction with regard to kinesthetic sensory modality was adopted. Subjects were blindfolded and asked to reproduce the criterion-movement (length of 10 cm) toward right hand side in space. Three different experimental spaces were used in relation to the subject's body positions; 'left space (range from left shoulder to median line)', 'right space (from median line to right shoulder)', and 'right outside space (outside of right shoulder)'. The criterion-movement was presented to the subjects, and then they reproduced the movement in either one of these three spaces. Generally, the left USN patients reproduced movements with more errors and less accuracy when the execution side was in the 'left space'. The results confirmed the observation that left USN patients showed more frequent response toward the right hand side direction in the 'left space'. The left USN patients with lesions in frontal and/or basal ganglia regions showed larger errors in this task compared to the left USN patient whose lesion is localized in the parietal region in the right hemisphere. PMID- 8230872 TI - [The differences in role expectations of adolescent men and women for various others]. AB - Everyday discourse patterns of adolescents were examined to clarify differences in their implicit expectations for various other people. Subjects were asked to complete 50 questionnaires on self-disclosing behaviors (Study 1) and which person in whom they would talk (Study 2) in everyday life. In both studies importance about each question is also scored by seven point scale. Total preferred percentage was calculated about each selected person. The results were as follows: same-sex friends were most frequently chosen as the person, to talk to while parents were selected for more important matters. Males comparatively tended to choose their father, while females their mother, to talk important matters with. Males had a tendency to talk about only important matters, while females tended to ignore importance. Males had a tendency to talk with girl friends about things they told no one else, while females tended to share with same-sex friends the same information as with boy friends. Gender differences in implicit role expectations for various others were discussed. PMID- 8230873 TI - [Comparative study of adolescent views on their parents and their self-reliance in Japan and the United States]. AB - Differences between Japanese adolescent's (127 males and 227 females) and American adolescent's views on their relationships with their parents were studied. Japanese male students showed the lowest scores on both the father-child and mother child emotional tie scales, but indicated the highest scores on both the paternal and maternal control scales. It was found that the Japanese students who showed strong emotional ties to their mothers also showed high scores in the scale of attachment to their fathers. American students tended to have stronger emotional ties to their fathers than did Japanese students. In both countries, female students tended to have stronger mother-child emotional ties than male students. In addition, Japanese female students tended to show low scores on the self-reliant scale. PMID- 8230874 TI - [The development of visual co-action in infancy]. AB - The purpose was to examine (1) whether infants are capable of imitation or not and (2) to assess their ability for information processing, specifically for visual perception. Twenty-one infants were presented four kinds of facial expressions (tongue protrusion, opened mouth, mouth protrusion and eye blink) at the ages of six, ten, and fourteen weeks. Infants' responses were videotaped and analyzed. Their behavior indicated that six-week-olds could not differentiate between the four stimuli. They could not imitate the facial expressions, because their perception did not differentiate the stimuli sufficiently yet. Responses by ten- and fourteen-week-olds indicated that their perceptual processes had developed sufficiently to differentiate stimuli, and make some matching behaviors with the processed information. Some matching responses, open mouth and mouth protrusion, were less frequent at fourteen weeks than at ten weeks. However, the overall matching responses were greater among fourteen than ten week olds. Apparently with the development of perceptual processes, matching responses also emerge. PMID- 8230875 TI - [Communication in problem solving by preschool children: effects of subject's familiarity with experimenter and task orientation of experimenter]. AB - The purpose was to examine effects of subject's familiarity with experimenter, and the task orientation of experimenter towards preschool children's communication in a problem solving situation. After subjects were given one of three different information about a target card, they were asked to identify the target one. However, subjects were unable to identify the target one without questions. The main results were as follows: (1) 5-year-olds made more questions and chose the target more often than 4-year-olds, (2) High familiarity group made questions and chose the target more often than low one, (3) Task-oriented group made more questions and chose the target more often than non-oriented one, and (4) Interaction of familiarity and the task-orientation was not significant. These results suggested that familiarity and task-orientation affected rate of communication occurrence in problem solving by preschool children. PMID- 8230876 TI - [Improvement of biventricular heart failure in a case of obstructive sleep apnea syndrome by nasal CPAP therapy]. AB - A 42-year-old male patient with obstructive sleep apnea syndrome (OSAS) suffering from biventricular heart failure is reported. He had been treated for OSAS with conventional therapy. However, he complained of severe dyspnea in association with extreme weight gain and general edema. Therefore, he was admitted to our department. He weighed 168 kg on admission, and marked edema was observed. Chest film revealed significant dilatation of the cardiac silhouette and pleural effusion. PaO2 was 37 mmHg and PaCO2 was 66 mmHg. Polysomnography showed an apnea index of 58.3 and severe oxygen desaturation during sleep. Right heart catheterization showed elevation of mean pulmonary artery pressure mPAP: 55 mmHg) and pulmonary capillary wedge pressure (Pcwp: 33 mmHg) suggesting biventricular heart failure. Digitalization and diuretic therapy were immediately initiated. In addition, nasal CPAP was applied to this patient during sleep, and sleep apnea and oxygen desaturation were almost completely reversed. Significant diuresis was observed, and blood gas data and sleep disturbance were improved. Fifty-nine days after admission, his weight had decreased to 96 kg, and mPAP and Pcwp decreased to 32 and 23 mmHg, respectively. This case demonstrates that nasal CPA is an effective tool for the treatment severe OSAS patients. PMID- 8230877 TI - [A case of intralobar pulmonary sequestration associated with an accessory diaphragm]. AB - We report one operated case (a 71-year-old female) of intralobar pulmonary sequestration (Pryce's type 1) associated with an accessory diaphragm. The patient had hemoptysis for 5 years. Lateral chest X-ray showed an oblique line attaching to the central portion of the right diaphragm. Selective angiography and ultrasonography showed an aberrant vessel originating from the abdominal aorta. At the time of operation, the pleural cavity was divided into two compartments by a fibrous membrane. The bronchus of the lower compartment and the pulmonary veins ran through a hole in the accessory diaphragm. The aberrant artery flowed into the parts of lung in the upper and lower compartments. PMID- 8230878 TI - [A case of surgically resected primary pulmonary lymphoma with IgG paraproteinemia: gene analysis was effective for establishing its diagnosis]. AB - A 69-year-old woman was hospitalized because of abnormal lung shadow and IgG lambda paraproteinemia. She was otherwise healthy and asymptomatic. Chest roentgenogram showed a consolidation in the right lower lobe. Chest CT showed a tumour in S8 and subpleural interstitial shadow in S10. No intrathoracic lymphadenopathy was found. Serum IgG was 6,109 mg/dl. The cell count obtained by bronchoalveolar lavage (BAL) was 44.5% plasma cells and 17.5% lymphocytes. CD19 positive lymphocytes were prominent. The IgG/albumin ratio was 13 times higher, and IL-6/albumin ratio was 29 times higher in lavage fluid than in serum. Transbronchial lung biopsy (TBLB) specimen showed interstitial infiltration of plasma cells and lymphocytes. Right lower lobectomy was performed, and serum IgG subsequently decreased to about 4,000 mg/dl. DNA was extracted from the surgical specimen, and analyzed by Polymerase Chain Reaction (PCR) method. A rearrangement band was amplified with Fr3a & VLJH primers (immunoglobulin heavy chain gene). The infiltrated cells were proved to be monoclonal B-cells. This case was diagnosed as small lymphocytic lymphoma, plasmacytoid. Most primary pulmonary lymphomas are well differentiated B-cell type, and the histopathological findings resemble those of LIP or pseudolymphoma. Gene analysis may thus be an effective procedure for the distinction between inflammatory and neoplastic cell proliferation, such as LIP and lymphoma. PMID- 8230879 TI - [A case of intrapulmonary neurilemmoma]. AB - A 33-year-old man was admitted to our hospital because of an abnormal shadow on chest X-ray film. Chest X-ray film and chest CT showed a round tumor in the left infrahilar area. Bronchoscopic findings revealed narrowing of left B5 and B8, with extraluminal compression. A preoperative diagnosis was not obtained and tumor resection was performed. The tumor was located in the parenchyma of left S8. Histological examination showed nuclear palisading and loosely textured spindle cells within myxomatous tissue, compatible with neurilemoma. Intrapulmonary neurilemoma is extremely rare and only 23 cases have been previously reported in Japan. PMID- 8230880 TI - [Bronchospasms caused by intravenous hydrocortisone sodium hemisuccinate (Solu Cortef) in an aspirin-sensitive asthmatic patient]. AB - This is a report of a male patient who had an anaphylaxia-like reaction to hydrocortisone sodium hemisuccinate (Solu-Cortef). A 32-year-old male with aspirin sensitivity was treated for moderate wheezing with 200 mg Solu-Cortef given intravenously. After this treatment, his condition deteriorated acutely and he was admitted to our hospital. Intradermal skin tests revealed positive immediate reactions to Solu-Cortef, Saxizon, Solu-Medrol and Predonine. Challenge tests with 100 mg Solu-Cortef and Saxizon injection showed 34% and 27% decrease, respectively in FEV1 15 min after injection. These results suggest that succinylation of steroids resulted in allergic reaction to steroids in this patient. PMID- 8230881 TI - [A case of pulmonary thromboembolism associated with heparin-induced thrombocytopenia]. AB - A 41-year-old man was admitted to our hospital for dyspnea on exertion and chest oppression. He was diagnosed as having pulmonary thromboembolism by pulmonary perfusion scan and angiography. An anticoagulant (heparin) and fibrinolytic (urokinase) therapy were administered to him as standard therapy. Six days after continuous infusion of heparin, however, he complained of worsening of dyspnea associated with severe hypoxemia and thrombocytopenia. Because heparin-induced thrombocytopenia was suspected, heparin administration was discontinued. Shortly after switching anticoagulant therapy from heparin to warfarin, the patient's symptom improved and his platelet count returned to the pretreatment value. In this case, it was thought that an immune-mediated response contributed to this heparin-induced thrombo-cytopenia. Moreover, we revealed platelet aggregating factor in the patient's plasma. This case illustrates that caution must be taken in the use of heparin for anticoagulant therapy in patients with pulmonary thromboembolism. PMID- 8230882 TI - [A case of sleep apnea syndrome with significant alteration of apnea index by sleep position]. AB - An obese 37-year-old man (130% ideal body weight) was admitted to our hospital with the complaint of excessive daytime sleepiness. During all-night polysomnography, he showed predominantly obstructive sleep apnea and an apnea index (AI) of 57.5, and was diagnosed as having obstructive sleep apnea syndrome (OSAS). However, AI values calculated separately for the time in the supine position (AI-S) and in the lateral decubitus position (AI-L) were 82.4 and 5.9, respectively. Moreover, duration of apnea, lowest SaO2, and the quality of sleep were improved when sleeping in the lateral compared to the supine position. After weight reduction of 7 kg, AI in total decreased to 33.2, although AI-S was 77.3 and AI-L was 3.8. The improvement of AI in total to be due to a relative increase during sleep in the lateral position. These results suggest that in some OSAS patients, the sleep position should be taken into account in assessing the severity of the disease or in evaluating the effects of therapy. This study also suggests the efficacy of sleep position adjustment in the treatment of OSAS. PMID- 8230883 TI - [A case of acute eosinophilic pneumonia with Kerley's lines (A and B) on chest X ray]. AB - A 18-year-old woman presented to our hospital complaining of an acute onset of progressive dyspnea with nonproductive cough and high fever. The patient was in her usual good health until the previous day, when she started to develop symptoms 8 hours after taking aspirin for a headache. The chest roentgenogram revealed Kerley's lines (A and B), perivascular cuffing and hilar haze with bilateral pleural effusions. Body temperature was 38 degrees C and PaO2 was 48 torr. Infectious diseases and extrinsic allergic alveolitis were excluded. The lymphocyte stimulating test was negative for aspirin. Acute eosinophilic pneumonia was strongly suggested by bronchoalveolar lavage showing a marked increase in eosinophils without peripheral eosinophilia. By the seventh hospital day all clinical and radiographic signs were improved without steroid therapy. Most cases of acute eosinophilic pneumonia reported previously showed diffuse infiltrative shadows on the chest roentgenogram. The present case had interesting radiographic findings which suggested interstitial pulmonary edema. PMID- 8230884 TI - [A case of metastatic iris tumor from lung cancer presenting as decreased vision]. AB - We presented a case of metastatic iris tumor from lung cancer first presenting as decreased vision. A 47-year-old male was referred to our hospital, complaining of decreased vision. At his first visit, a yellowish mass was found on the right iris, and chest X-ray revealed paratracheal, subcarina, and right hilar lymphadenopathy. Bronchoscopy was performed, and small cell carcinoma was detected in a transbronchial biopsy specimen. Sixteen cases of metastatic iris tumor from lung cancer have been reported of such rare occurrences in Japan. The clinical features are briefly reviewed. PMID- 8230885 TI - [Peripheral polyneuropathy due to sarcoidosis in a patient with intrathoracic, ocular and skin lesions]. AB - Sarcoidosis is a multisystem disorder of unknown etiology presenting most frequently with bilateral hilar lymphadenopathy (BHL), pulmonary infiltration, and ocular and skin lesions. Neurological manifestations are found in about 5% of patients with systemic sarcoidosis. We report the case of a 58-year-old woman with neurosarcoidosis manifesting as isolated, peripheral polyneuropathy. This is a rare neurological manifestation. Two years before admission, she was found to have an abnormal chest radiograph showing BHL, and pulmonary sarcoidosis was diagnosed by lung biopsy. Six months later, she noted facial erythematous lesions, and lupus pernio was confirmed by skin biopsy. Fifteen months before admission, she developed visual disturbance of the right eye caused by sarcoid uveitis. Two months before admission, she complained of paresthesia and weakness of limbs. She was admitted for nerve-biopsy. Sural nerve biopsy revealed sarcoid granulomas in the epineurial space, periangitis and axonal degeneration. Neurologically, the diagnosis of mononeuritis multiplex without cranial nerve palsy was made. Treatment with 30 mg prednisolone daily was initiated, and neurological and extraneurological improvement was noticeable within two weeks of treatment. The dose was carefully lowered over several months. PMID- 8230886 TI - [Metastatic lung cancer presenting with multiple thin-walled cavity-like shadows in a young adult with gallbladder cancer]. AB - A rare case of metastatic lung cancer presenting with multiple thin-walled cavity like shadows in a young adult with gallbladder cancer is reported. A 30-year-old man consulted our hospital with fever, cough, and general malaise. His chest X ray film and computed tomogram showed multiple nodular shadows and thin-walled cavity-like shadows. Cytology of sputum and bronchoalveolar lavage fluid showed class V (adenocarcinoma). Although two cycles of systemic combination chemotherapy (CDDP+VDS+MMC) were performed, his lung cancer progressed. Finally, he died of obstructive jaundice from lymph node metastasis at the pancreatic head. At autopsy, the primary site of adenocarcinoma was found to be the gallbladder. Metastatic lung tumor from gallbladder cancer is common, and thin walled cavity-like metastasis to the lung in a young adult is a rare occurrence. PMID- 8230887 TI - [Three-dimensional CT images of the lung--preliminary study using inflated lung specimens]. AB - Three-dimensional CT images of six inflated lung specimens (4 lung cancer, 1 bronchiectasis, 1 hamartoma) were reconstructed from contiguous serial sections to study some probable problems in the clinical application of the technique. Optimal thickness and space of sections were evaluated, and contiguous 1.5-mm serial sections were found to provide a clear 3-D image with high fidelity. It was difficult to set a proper threshold to create clear images because a wide CT window is inconsistent with low noise. The images of tubular and linear structures such as from bronchi and vessels demonstrated higher resolution when reconstructed from scanning in the more perpendicular cross-sectional direction to their axis. Although 3-D images obtained here were not as clear as expected, technological advances such as helical CT or MRI may provide 3-D images with sufficient resolution to be applied clinically. PMID- 8230888 TI - [A comparison of lymphocyte stimulation test results and challenge test results in 19 cases of antituberculous drug-induced allergy]. AB - We clinically investigated 19 cases of antituberculous drug-induced allergy. The results of the challenge tests were positive in 30 out of the 55 drugs tested. The results of the lymphocyte stimulation tests were positive in 19 out of the 55 drugs tested. In 84.2% of the cases with positive lymphocyte stimulation test findings, the challenge test was also positive. It can be concluded from these results that the lymphocyte stimulation test may be useful in the detection of the causes of drug induced allergies in patients undergoing drug therapy for tuberculosis. PMID- 8230889 TI - [Measurement of myeloperoxidase and thiobarbituric acid-reactive material in plasma and bronchoalveolar lavage in E. coli-induced acute lung injury]. AB - Myeloperoxidase (MPO), which is exclusively contained in neutrophils, is released on their activation. Therefore, MPO may possibly be used as a parameter of neutrophil activation. Thiobarbituric acid-reactive material (TBARM) reflects lipid peroxidation and is a parameter of oxygen radical-mediated cell membrane damage. Using our guinea pig model of septic lung injury we measured MPO and TBARM in the setting of acute lung injury. The two experimental groups were saline controls (n = 8) and an E. coli septic group to which 2 x 10(9) live E. coli were administered intravenously (n = 8). Lung damage was assessed by measuring wet to dry lung weight ratio (W/D) and lung tissue to plasma accumulation of 125I-albumin (AL: albumin leakage). We measured MPO and TBARM in plasma and BAL fluid. Increased W/D and AL were observed in the E. coli group suggesting the development of acute lung injury. In the E. coli group, plasma MPO increased and MPO in BAL fluid was significantly increased as compared with the saline control group. There was no difference in plasma TBARM between the two groups, while TBARM in BAL fluid of the E. coli group was greater than in that of controls. Although BAL fluid TBARM correlated with both W/D and AL, there was no relation between BAL fluid MPO and either of these parameters. We conclude that TBARM in BAL fluid may be useful for assessing E. coli-induced acute lung injury in guinea pigs. PMID- 8230890 TI - [Serum soluble CD23 and soluble CD25 in bronchial asthmatic patients]. AB - Serum sCD23 and sCD25 levels in asthmatic patients and healthy subjects were assayed. Serum sCD23 levels in asthmatic patients were significantly higher than those in healthy subjects (p < 0.05). Serum sCD25 levels in asthmatic patients were also significantly higher than those in healthy subjects (p < 0.02). There was a significant correlation between serum sCD23 and sCD25 levels in asthmatic patients (r = 0.52, p < 0.001). It is well known that sCD23 and sCD25 levels are regulated by different cytokines, and this study suggests that T cells play important roles in the pathogenesis of asthma. PMID- 8230891 TI - [Effects of cilostazol, a cyclic AMP phosphodiesterase inhibitor, on pulmonary vascular tone]. AB - We examined the effect of Cilostazol, a cyclic AMP phosphodiesterase inhibitor, rat isolated pulmonary arterial ring tone. Cilostazol dilated pulmonary arterial rings pre-contracted with 10(-6) M phenylephrine in a dose-dependent manner (ED50 3.00 x 10(-6) M). This vasodilatory effect of Cilostazol was not affected by pretreatment with meclofenamate (10(-5) M), mechanical endothelium denudation, methylene blue (10(-5) M) or nitro-L-arginine (2 x 10(-4) M). The vasodilatory effect of Cilostazol on rat thoracic aortic rings was stronger than that on rat pulmonary arterial ring (ED50 1.89 x 10(-6) M). Cilostazol (10(-6) M-10(-4) M) inhibited hypoxic contraction of rat pulmonary rings in a dose-dependent manner. Our experimental data indicated that Cilostazol caused vasodilation regardless of vascular endothelium function and inhibited hypoxic contraction due to inhibition of cyclic AMP phosphodiesterase. PMID- 8230892 TI - [Pulmonary macrophages in rats after hemithoracic irradiation: analysis of morphology and expression of surface antigen]. AB - The pathogenesis of radiation pneumonitis is not completely understood. The long latent period and involvement of unirradiated lung tissue may indicate an immune reaction in the injurious process of irradiated lung. To investigate the role of pulmonary macrophages in radiation pneumonitis, morphology and membrane antigen expression of pulmonary macrophages were studied in irradiated rat lung tissue following 4000 R hemithoracic irradiation. Lungs were explanted at 2, 4, 6, 8, 16, and 28 weeks after irradiation. Cryosections of irradiated lung tissue were immunohistochemically studied, and alveolar macrophages in bronchoalveolar lavage fluid were also analyzed using monoclonal antibodies to rat major histocompatibility complex (MHC) antigens and macrophages with flow cytometry. Macrophage subpopulations were analyzed using a panel of monoclonal antibodies to MHC class I (HAM2), MHC class II (OX6) and macrophage differentiation antigens (ED1, ED2, ED3). Alveolar macrophages in bronchoalveolar lavage were morphologically studied by smear and flow cytometry of forward light scatter and 90 degrees light scatter. At 2 weeks after irradiation, when histological changes did not appear, small lymphocyte-like macrophages and large foamy macrophages were observed in both the smear and histograms by flow cytometry. At 4, 6, and 8 weeks after irradiation, these new populations had markedly increased. However, at 16 and 28 weeks after irradiation, the size and shape of alveolar macrophages had returned to normal. In the expression of macrophage membrane antigens, an increase in the frequency of MHC class II+ cells in lavaged cells appeared at 2 weeks after irradiation, and became significant at 4 weeks.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230893 TI - [Clinical and pathological studies of organizing pneumonia]. AB - Clinical and pathological studies on organizing pneumonia (OP) were performed in 16 cases diagnosed by transbronchial lung biopsy (TBLB). All patients were elderly and poorly-nourished with underlying disease, and suffered from fever and various respiratory symptoms. Results of laboratory studies in these cases were positive for C-reactive protein, negative for tuberculin reaction and showed high complement levels. Pathological findings demonstrated that there were two kinds of organizing process classified according to whether fibrin exudation was detected in Masson bodies or not. Twelve of the 16 cases were treated with prednisolone, and two cases were observed without administration. In nine of the 16 cases, abnormal shadows in chest X-ray disappeared, although abnormal shadows remained in five. Regarding the relationship between pathological findings and shadows in chest X-ray, Masson bodies without fibrin were observed in nine cases in which abnormal shadows in chest X-ray disappeared, but Masson bodies containing fibrin were observed in five cases in which abnormal shadows remained. These results suggest that there are two kinds of organizing process in OP. Steroid therapy tends to be ineffective in OP demonstrating organization containing fibrin caused probably by infection. However, prednisolone is effective in the OP associated with a fibrosing process, similar to idiopathic BOOP, without or with unrelated fibrin exudation of unknown origin. PMID- 8230894 TI - [Three-dimensional computed tomography for the evaluation of peripheral small lung cancers]. AB - To facilitate the accurate diagnosis of small peripheral lung cancers (less than 30 mm in diameter), we evaluated the use of three-dimensional computed tomography (3D-CT). Conventional CT images (10 mm thick), thin-slice CT images (2 mm thick) and 3-D-CT images were prospectively studied in 29 patients with peripheral lung cancer (23 adenocarcinoma, 3 squamous cell carcinoma, 1 large cell carcinoma, 1 adeno-squamous cell carcinoma, 1 small cell carcinoma) who underwent thoracotomy. Patients were 20 males and nine females with a median age of 65 (range 38-75) years. Conventional CT and thin-slice CT images were examined with an extended scale (window level: -600, window width: 1900) using high resolution images. 3D CT images were made using TCT-900S (Toshiba, Tokyo) software. In resected cases, surgical specimens were sliced transversely to correlate with the CT images. Comparison of diagnoses based on CT findings and on pathological findings showed that 3D-CT was 96% accurate for detecting involved vessels, while conventional CT was 47% and thin-slice CT was 87% accurate. Thus 3D-CT was superior to conventional CT for detecting involved vessel (p < 0.01). 3D-CT images also showed specific features of the lesions, including the appearance of the margin and pleural indentations, as seen in thin-slice CT images. These results suggest that 3D-CT images may have an important role in diagnosing small peripheral lung cancers by demonstrating involved vessels and the morphology of the lesions. PMID- 8230895 TI - [Relationships between emphysema, bronchiolar lesions, and pulmonary function tests in mild centrilobular emphysema of the human lungs]. AB - The relationships between emphysema, bronchiolar lesions, and pulmonary function tests were studied in very mild centrilobular emphysematous lungs. The severity of emphysema was significantly related to VC, FEV1, FEV1/FVC, the slope of phase III, CC/TLC and static elastic lung recoil. Emphysema was not correlated with DLco. Bronchiolar lesions were poorly correlated with pulmonary function tests. Emphysema was related to bronchiolar lesions, such as smooth muscle hypertrophy and goblet cell metaplasia. These findings suggest that alterations of the mechanical property of the lung occur in very mild centrilobular emphysema. Measurements of these abnormalities may be useful to detect early emphysema. PMID- 8230896 TI - [Amlexanox has an acute bronchodilator effect in patients with aspirin-induced asthma (AIA)]. AB - Recently, we have reported that a nebulized solution of sodium cromoglycate (SCG) (Intal), a well known antiallergic agent, has an acute bronchodilator effect in patients with aspirin-induced asthma (AIA). Amlexanox is an antiallergic agent, with a structure similar to that of SCG. Therefore, we investigated the acute bronchodilator effect of amlexanox administered orally to adult asthmatics, focusing on a comparison between AIA and non-AIA patients. Fifteen patients participated, 8 having AIA and 7 non-AIA. After performing spirometry during remission, they were administered either amlexanox or placebo using a randomized double-blind protocol. Spirometry was then performed at 30, 60, 90 min, 2 h, and 3 h after administration. The placebo used was lactose. In the AIA group, FEV1 was significantly improved after administration of amlexanox. However, FEV1 was significantly decreased after placebo administration. On the other hand, in the non-AIA group, FEV1 was not changed after administration of either of the drugs. From these results, we conclude that amlexanox has an acute bronchodilator effect only in AIA patients. PMID- 8230897 TI - [Clinical studies in three cases of pulmonary cryptococcosis]. AB - We report three cases of pulmonary cryptococcosis in which one community acquired and two nosocomial infections were suspected. Clinical studies were focused on histological and immunological diagnosis and antifungal chemotherapy. The first case was initially suspected of having primary cryptococcosis. The second case was first suspected to have BOOP on the basis of clinical and radiological findings. The third was initially suspected of having lung cancer because of an enlarging mass lesion in fibrotic lung and elevated tumor markers. All cases were treated with antifungal agents. Two cases were treated with fluconazole alone, the other case with fluconazole and 5-fluorocytosine. In all cases, the abnormal shadows on chest X-ray demonstrated 50 to 90 percent improvement within 6 months, and cryptococcal antigen in the serum decreased. There were no side effects from the drugs. PMID- 8230898 TI - [Successful treatment of diaphragm pacing-induced obstructive sleep apnea syndrome with nasal CPAP]. AB - We report a 71-year-old female patient with primary alveolar hypoventilation syndrome who received diaphragm pacing (DP) and developed obstructive sleep apnea syndrome (OSAS). Application of nCPAP markedly improved her nocturnal hypoxemia. The monitored polygrams before and after the application strongly suggested that the main mechanism of OSAS was an imbalance of activity between upper airway dilator muscles and pump muscles. Moreover, paradoxical movement of the rib cage is not necessarily due to upper airway obstruction. Monitoring of tidal volume and arterial oxygen saturation is essential for the diagnosis of DP-induced OSAS. PMID- 8230899 TI - [A case of chronic asthma with emphysematous change in a middle-aged woman]. AB - A 32-year-old woman with chronic asthma is presented. She had suffered from dyspnea and breathlessness on exertion for 7 years. Her chest X-ray film on admission showed marked emphysematous changes and chest CT scan revealed low attenuation areas, however she was a non-smoker and had no previous childhood respiratory problems. She was administered prednisolone to treat the wheezing attack and hypoxemia. After treatment, her lung function and hypoxemia improved, but low attenuation areas of the chest CT scan remained unchanged. These facts suggest the possibility that bronchial asthma which is controlled poorly for a long time, may cause emphysematous change of the lung. PMID- 8230900 TI - [Pacemaker implantations in children with regard to the site of implantation]. AB - We reviewed thirty eight children, ranged 3 days to 15 years (mean, 7.2 years) of age, who underwent permanent pacemaker implantations in our hospital. Long-term results including the site of generator pocket were discussed. They are alive and well except two early and three late deaths that were not related to the pacemaker implantation. Twenty-seven epicardial and 11 endocardial electrodes were implanted at the initial operations. Thirteen pulse generators implanted in subaxillar position had no complications, otherwise, 12 subcostal implantation resulted 4 skin necrosis and 3 lead fractures. Eleven anterior chest implantations used for relatively older children resulted in one skin necrosis. We recommend subaxillar implantation for children, especially for small infants. The replacements of pulse generators due to battery depletion were done between 1.7 to 5.5 years (mean 4.0 years). Because epicardial implanted electrodes often show higher stimulation threshold than epicardial ones, pacing rate should be as lowered as possible if the patient had no symptom. Stab-in leads placed on the atrial wall sometimes caused the elevation of stimulation threshold and undersensing. We conclude that the development of more reliable and stable electrode is desired to improve the long term results of pacemaker therapy in children although the improvement of pulse generator itself and the operative procedure. PMID- 8230901 TI - [Treatment of thoracic malignancy accompanied with cardiovascular disease]. AB - We treated 10 cases of thoracic malignancy accompanied with cardiovascular disease. Among thoracic malignancy, 7 cases were lung cancer and 3 were esophageal cancer. Accompanied cardiovascular diseases were ischemic heart disease (2 cases), valvular disease (3 cases), WPW syndrome (1 case), aortic aneurysm (4 cases). The mean age was 66, ranged from 51 to 79. The simultaneous occurrence of the two lesions were observed in 6 cases and thoracic malignancy was diagnosed after a varying interval of time following surgery of cardiovascular disease in 4 cases. In cases of thoracic malignancy accompanied with heart disease, the treatment of heart disease should precede the operation of malignant disease to reduce the risk of surgery. For the patient with esophageal cancer, posterior mediastinal esophagostomy should be applied who may have heart surgery in future. In cases of coexisting malignancy and aortic aneurysm, the priority of treatment should be determined considering the size of aneurysm. If the transverse diameter of aneurysm is larger than 7 cm, there is a high risk of rupture, so surgery for the aneurysm precedes operation of malignant diseases. It is desirable to avoid concomitant operation of malignancy and cardiovascular disease. PMID- 8230902 TI - [Study on extravascular lung water and polymorphonuclear leukocyte elastase during acute phase following radical treatment of lung cancer: effect of ulinastatin on respiratory functions]. AB - We studied respiratory functions, extravascular lung water (EVLW) as measured by a thermodilution method using double Na indicators, and variations in polymorphonuclear leukocyte elastase (PMNE) during the acute phase following radical treatment of lung cancer, as well as to investigate clinical effect of ulinastatin. Using 16 patients with lung cancer who had undergone surgery of R 2 a or more, 10 patients without ulinastatin therapy were compared with six patients given 300,000 U/day of this drug by drip infusion from Day 0 and 4 of disease. It was found that EVLW value decreased most on Day 1, and gradually increased thereafter but did not return to the preoperative level. PMNE value increased from immediately after surgery and peaked on Day 3, but returned nearly to the preoperative level by Day 5. Although this was different from the variations in EVLW value, there was a correlation between these two parameters on Day 3 when PMNE peaked. Further, ulinastatin tended to inhibit PMNE and improve oxygenation index. Respiratory and circulatory parameters vary in complex manners during the acute phase following radical treatment of lung cancer, and could not be accurately evaluated by means of thermodilution using Na. PMNE is considered a contributing factor to pulmonary tissue disorders following radical treatment of lung cancer, and ulinastatin appeared to be effective in the postoperative management of patients undergoing radical treatment of lung cancer. PMID- 8230903 TI - [Pre and postoperative left ventricular volume and function in infants and children with total anomalous pulmonary venous return]. AB - From January, 1989, to December, 1991, 36 consecutive patients with total anomalous pulmonary venous return (TAPVR) underwent intracardiac repair (ICR) in our institute. We studied the relationship between the size of atrial septal defect (ASD), age and preoperative left ventricular end-diastolic diameter (LVEDD) in 28 patients with regression analysis. There was no statistical relationship between the size of ASD, age and LVEDD. We measured LVEDD, RV to LV pressure ratio (RVP/LVP) and LV fractional shortening (LVFS) using two dimensional echocardiography in 15 patients, prior to and following ICR. LVEDD increased from 1.16 +/- 0.40 cm to 1.62 +/- 0.28 cm immediately following ICR, and then increased to 1.79 +/- 0.34 cm one month following ICR again. LVFS decreased from 38% to 35% immediately following ICR but still remained within normal range (normal range 28-45%). RVP/LVP decreased from 1.0 to 0.69 immediately following ICR, and to 0.39 one month following ICR. We concluded that the decrease of preoperative LV end-diastolic volume in TAPVR was not due to the decrease of LV preload but to anterior deviation of the ventricular septum caused by high RV pressure. There was significant increase of LV volume immediately following ICR, but LVFS maintained in normal range. This study suggested that small LV volume in patients with TAPVR did not influence the surgical results. PMID- 8230904 TI - [Long-term result of surgical patients with ischemic heart disease 70 years of age and older]. AB - This is a report of long-term follow-up of patients 70 years of age and older who underwent operations for ischemic heart diseases. (1) Two of them died in this follow-up period, one due to pneumonia seven months postoperatively, another due to lung cancer 20 months after operation. Actuarial survival rate was 96.8% at one year, 93.3% between two and five years. (2) Event-free rate was 80.6% at one year, 70.1% at two years, and 61.1% between three and five years. (2) Although 21 patients were classified in grade 0 (normal activity) of performance status, seven were classified between grade 2 and 4. We concluded that long-term results were good in surgical patients 70 years of age and older with ischemic heart disease. PMID- 8230905 TI - [Preoperative evaluation of a distal aortic arch aneurysm by spiral CT with three dimensional reconstruction]. AB - Sixty-seven-year-old male with a distal aortic arch aneurysm was evaluated by spiral CT with three-dimensional reconstruction. Spiral CT allows rapid data acquisition through an entire anatomic region during a single breath hold. The helically acquired data undergo interpolation. Transverse images are reconstructed with slice intervals 1-4 mm apart. Three-dimensional images were generated by these data with the transverse images. The entire anatomical correlation with the aneurysm and the aortic arch branches was clearly recognized by these three-dimensional images. Preoperative evaluation of the distal aortic arch aneurysm by spiral CT with three-dimensional reconstruction was useful for the surgical treatment. PMID- 8230906 TI - [Extraction of infected pacemaker electrode using catheter and snare]. AB - A 85-year-old man had undergone implantation of a transvenous pacemaker system in 1982. Nine years later the generator was explanted and he was underwent implantation of a new generator and electrode. After one year of the second operation, infection of the old electrode had been noted and it was not cured in spite of partial explantation of the infected electrode and appropriate antibiotic therapy. We decided to remove the infected electrode and succeeded in extraction using catheter and snare. The patient's recovery was uneventful. We suggest catheter and snare is useful for explantation of chronic transvenous pacemaker electrodes. PMID- 8230907 TI - [A case of intrathoracic dumb-bell ganglioneuroma and a surgical approach using spinal evoked potentials (SEP)]. AB - A successfully treated case of a seven-year-old girl with a left intrathoracic Dumb-bell ganglioneuroma is reported. The tumor was 10 cm in diameter. Preoperative angiography revealed that the location of tumor was very close to the Adamkiewicz artery. In order to prevent the artery from damaging, spinal evoked potentials (SEP) was used during the surgery and was very useful for monitoring the spinal cord. Although SEP has often been employed in spine surgery, the literature on the use of it for posterior mediastinal tumor is scarce. It is considered that SEP is very useful for not damaging the feeding artery to the spinal cord, and for preventing the postoperative neurologic complications. PMID- 8230908 TI - [Transvenous monitoring of left atrial and central venous pressure with a double lumen catheter]. AB - We developed a new method to monitor left atrial pressure in the patients undergoing open heart surgery. The advantages of our method are 1) freedom from complications relating its removal (eg. bleeding) because of its transvenous insertion, and 2) simultaneous monitoring of both right and left atrial pressures by a single catheter. We designed to place the second port of a two-lumen central venous catheter 5 or 7 cm proximal to the tip. The catheter is inserted into the right atrium before surgery and the tip is advanced in the left atrium through the foramen ovale intraoperatively. We used this catheter in 16 infants and children, and found that effective hemodynamic monitoring was obtained without any complications. We recommend this method for the postoperative management of open heart surgery in the pediatric patients. PMID- 8230909 TI - [In vitro evaluation of prostaglandin I2 analogue on hypothermic injury to immature myocytes]. AB - We evaluated the functional and biochemical effects of prostaglandin I2 analogue (PGI-A) on cardiac myocytes incubated under hypothermic conditions. Myocytes were isolated from neonatal rat ventricles and cultured for 4 days. Then, myocytes (12.5 x 10(5) myocytes/flask) were incubated at 4 degrees C for 24 hrs in media with PGI-A as follows; O M PGI-A (group C: control), 10(-9) M (group P1), 10(-8) M (group P2), 10(-7) M (group P3), 10(-6) M (group P4). After hypothermic incubation, CPK and LDH were measured. The myocytes were then cultured for 24 hrs at 37 degrees C to evaluate the recovery of myocyte beating rate. For the beating rate, group P2 showed significantly increased recovery compared to the control (P2: 47.9, p < 0.025, C:18.1 percent of control; ie, beating rate prior to hypothermic incubation). The release of CPK and LDH was significantly suppressed in group P2 compared to the control (P2:57.7, p < 0.05, 275.1, p < 0.025; C:96.8 mIU/flask, 439.6 mIU/flask, respectively). In conclusion, prostaglandin I2 has direct cytoprotective characteristics for immature myocytes that may be suitable for cardiac preservation. PMID- 8230910 TI - [A case of operated invasive thymoma accompanied with the thrombocytopenia]. AB - Thymoma with thrombocytopenia has rarely been reported. We recently encountered a case of operated invasive thymoma with thrombocytopenia. A 74-year-old woman was admitted to our hospital with the chief complaint of 'shortness of breath' and general fatigue. Chest X-ray, CT and MRI showed a mass 5 x 5 cm in size at the anterior mediastinum. In hematological test, a significant decrease in platelets count (1.6 x 10(4)/mm3) and a increase in PA-IgG (134.5 ng/10 cells) were found. This finding suggests that thrombocytopenia is due to immunological abnormality. Steroid therapy (prednisolone 60 mg) was begun but was not effective and then, extended thymectomy was performed. After operation, prednisolone (30 mg) therapy was continued. Although prednisolone therapy was gradually decreased and stopped, platelets count was gradually increased 10.8 x 10(4)/mm3. Extended thymectomy and steroid therapy were effective. PMID- 8230911 TI - [Coronary artery bypass grafting in six patients with familial hypercholesterolemia]. AB - From June 1989 through November 1991, six patients with familial hypercholesterolemia underwent surgical treatments. The surgical procedures were coronary artery bypass grafting (CABG) alone 4, ligation of coronary aneurysm + CABG 1, and CABG + femoro-femoral bypass 1. There were no operative or hospital deaths. Early post operative patency rate of the grafts was 100%. CASE PRESENTATION: A 44-year-old man whose anginal pain recurred 11 years after CABG. Coronary angiogram revealed stenosis and a large aneurysm in the circumflex coronary artery. Previous grafts to the left anterior descending coronary artery and diagonal branch were patent. Ligation of the aneurysm and internal mammary artery bypass grafting were performed. Postoperatively, the patient has remained asymptomatic. Our data indicate that CABG for FH patients is effective in the early postoperative period. PMID- 8230912 TI - [Successful treatment by using a pedicled omental flap for sternal osteomyelitis and mediastinal infection caused by MRSA]. AB - A 56-year-old male case, who was complicated with sternal osteomyelitis and mediastinitis after aortic valve replacement, was reported. Its infection was caused by MRSA (methicillin resistant Staphylococcus aureus). He was treated with sterilization of wound using povidone iodine, Minomycin solution and its injection for 10 days. And thereafter we decided to apply the plastic surgical technique to this case. After debridement of the infected granulation and sternum thoroughly, a pedicled omental flap was transposed successfully. PMID- 8230913 TI - [A case report of pseudocoarctation associated with aneurysm of the distal aortic arch]. AB - A 42-year-old female was admitted to our hospital for abnormal shadow in the left superior mediastinum on chest X-ray. Her blood pressure was 120/70 mmHg in the left arm and 134/72 in the right. Aortography revealed pseudocoarctation of the aortic arch distal to the left carotid artery with two aneurysm, and showed elongation between the left carotid artery and the left subclavian artery. Aneurysmectomy and graft replacement using a woven Dacron graft were successfully performed under cardio-pulmonary bypass. In 1969, Gay et al reported a case of pseudocoarctation which led to a fatal rupture of the thoracic aorta, so we recommend surgical therapy in cases of pseudocoarctation associated with aneurysm formation. PMID- 8230914 TI - [A case of Rendu-Osler-Weber disease associated with pulmonary arteriovenous fistula]. AB - A 60-year-old female was admitted with a chief complaint of left hemothorax. Chest X-ray films revealed an abnormal shadow when the pleural effusion disappeared. On the physical examination, multiple telangiectases were found at the nasal mucosa, tongue and stomach. The family history showed that her son experienced frequent episodes of epistaxis. The patient was diagnosed to have Rendu-Osler-Weber disease. CT scanning and pulmonary angiography showed an arteriovenous fistula of the left lower lobe (S10) and cerebral angiography did a small arteriovenous fistula in the cerebellum. Pulmonary partial resection was performed to prevent re-hemorrhage. Pulmonary arteriovenous fistula was 2.5 x 1.5 cm in size. It located in the left lower lobe exposing through the surface of the lung over the diaphragm. The postoperative course was uneventful and she was discharged on the 18th postoperative day without any complications. PMID- 8230915 TI - [Tetralogy of Fallot with aortopulmonary window: report of a successful repair]. AB - Tetralogy of Fallot is rarely associated with aortopulmonary window. A 6-month old boy with tetralogy of Fallot, aortopulmonary window, pulmonary atresia, patent left superior vena cava and right aortic arch was operated successfully. Both great arteries were divided, the defect in the aortic wall was closed directly, and the defect in the pulmonary arterial wall was connected with a valved external conduit which was used for right ventricular outflow tract reconstruction. Although oliguria had been prolonged for 3 weeks post operatively, he recovered completely and RV/LV pressure ratio was less than 0.6 estimated by two-dimensional echocardiography at one month after operation. PMID- 8230916 TI - [Successful surgical treatment of patent ductus arteriosus in adults: report of three cases]. AB - Three elderly patients underwent surgery for patent ductus arteriosus successfully. In two patients without calcification of ductus arteriosus, its division through a left thoracotomy was performed. In another patient with massively calcified ductus arteriosus associated with pulmonary hypertension, transaortic patch-closure of the aortic end of the ductus arteriosus was performed under partial cardiopulmonary bypass. All three patients and uneventful postoperative course. PMID- 8230917 TI - [A case report of BWG syndrome in an elderly patient performed with mitral valve replacement 11 years after single CABG]. AB - A 46-year-old female who had undergone single CABG for BWG syndrome 11 years ago was referred to our hospital for paroxysmal atrial fibrillation. The roentgenogram showed slightly cardiomegaly and the enlargement of LA. Ischemic changes of ECG appeared at I, aVL, V5, V6, as inverted T wave, and low voltage R wave at V1-V4, but non Q wave. The thallium-201 emission computed tomogram at exercise revealed poor perfusion at apical region without redistribution pattern. Catheterization showed mitral regurgitation (grade III), big right coronary artery (RCA) arising from aorta, rich collateral to poor left coronary artery (LCA), and bypass graft was obstructed. The proximal end of LCA was closed, and didn't arise from both pulmonary artery and ascending aorta. In this cases, MVR only without re-CABG to LCA was selected and performed. Postoperative course was uneventful. The result of this case suggested that MVR was an effective surgical procedure for MR of BWG syndrome in the adult case and it was better to add CABG to LCA as much as possible if the ischemic region was large. PMID- 8230918 TI - [A case of infective endocarditis with subaortic-valve aneurysm]. AB - We report a 64-year-old female patient with a subaortic-valve aneurysm due to infective endocarditis, in whom the aneurysm and its perforation was detected by Doppler color flow imaging. She had severe aortic and mitral valve regurgitation, and underwent aortic valve replacement with plication of the aneurysm. Transthoracic echocardiography doesn't have enough usefulness to clarify periaortic-annular changes, but combination with Doppler color flow imaging is supposed to be informative for subaortic-valve changes same as transesophageal echocardiography especially in the cases with abnormal flow. PMID- 8230919 TI - [A case of inflammatory pseudotumor of the lung: relation to metastasizing leiomyoma]. AB - A 61-year-old woman was referred to our clinic because of an abnormal shadow in the left lower lung field on an X-ray taken at a regular checkup. She had no subjective symptoms. Past history included myoma uteri at 47 yr and iatrogenic left pneumothorax subsequent to acupuncture for muscle pain in the shoulder. On CT, a solitary round lesion 2.5 cm in diameter and smooth margin was located at S5 just below the visceral pleura. Since we were unable to obtain a histological diagnosis by bronchofiberscopy, thoracotomy was carried out and the lesion was resected. The intraoperative pathological diagnosis was a benign lesion resembling leiomyoma. After surgery, re-evaluation of the myoma uteri specimen resected 14 years previously was done, as well as desmin immunohistochemical staining of the lung lesion, which demonstrated no evidence of a smooth muscle component in the latter. The final diagnosis was inflammatory pseudotumor. PMID- 8230920 TI - [A case of postoperative chylothorax successfully treated by chemical pleurodesis with OK-432]. AB - Recently, cases of postoperative chylothorax have increased concomitantly with the increase in number of aggressive thoracic surgery. Chylothorax was complicated on the 6 th postoperative day in a 69-year-old male who underwent right lower lobectomy with mediastinal lymph node dissection for adenocarcinoma of the lung. After the leaking point from thoracic duct was confirmed by lymphangiography and chest CT, OK-432 1.5 KE was instilled into the intrapleural space through a intrathoracic tube. By this procedure, the chylous fistulas were completely cured. Application of OK-432, even in such a small dose as 1.5 KE, appeared very useful for the treatment of postoperative chylothorax. PMID- 8230921 TI - [A case of mucinous cystic adenocarcinoma of the lung with signet-ring cells]. AB - A 70-year-old male case of mucinous cystic adenocarcinoma of the lung with signet ring cells was reported. Patient was operated with diagnosis suspicious lung cancer by the specimen from TBLB. But, intraoperative rapid pathological diagnosis was confused us for no cellular component, finally we performed right upper lobectomy and R1 LN dissection. Postoperative histological examination showed that the tumor was entirely filled with mucin and there are a few tumor cells with the signet-ring shape in its marginal area. So, this tumor was diagnosed as mucinous cystic adenocarcinoma of the lung with signet ring cells. This type of the lung adenocarcinoma was rare case, but we need to have much attentions because of the clinical differences from the other type of adenocarcinoma. PMID- 8230922 TI - [Surgical analysis for lung cancer with scalene node metastasis]. AB - In this study, we evaluated 15 surgically treated cases of lung cancer with scalene node metastasis. Eleven males and 4 females with ages ranging from 41 years to 78 years (average 59.1 years) were examined. Seven cases was in the right upper lobe, 3 in the left upper lobe, 2 in the right lower lobe, 2 in the left lower lobe, 1 in the middle lobe. Adenocarcinomas were found in 9 cases squamous cell carcinomas were in 4 cases and large cell carcinomas were in 2 cases. Two cases was in T 1, 7 in T 2, 2 in T 3, 4 in T 4. In all cases, lobectomy was performed. Five year survival rate was 14.3% as determined by the Kaplan-Meier method. Average survival period was 26.6 months. Three cases survived over 3 years including one case survived over 10 years. PMID- 8230923 TI - [Thirty-eight operative cases of mediastinal cyst: with particular reference to bronchogenic cyst]. AB - Thirty-eight patients (16 males and 22 females) with mediastinal cysts have been treated at the Second Department of Surgery at Okayama University from 1978 to 1989. There were thirteen cases of thymic cyst, nine cases of pericardial cyst one case of pericardial diverticulum, eight cases of bronchogenic cyst, two cases of dermoid cyst, two cases of cystic lymphangioma, one case of esophageal cyst, one case of gastroenteric cyst, and one case of aneurysmal bone cyst. MRI seemed to be useful for the qualitative diagnosis of mediastinal tumors as cystic or solid. Based on Maier's classification and considering the result of this study, bronchogenic cysts may be divided into the following groups: 1) paratracheal, 2) carinal, 3) hilar, 4) paraesophageal [a) upper (Iu), b) lower (Im, Ei)], 5) cephalad recess of the azygos vein (CRAzV) and 6) miscellaneous. According to this new classification, these bronchogenic cysts could be allocated to (3) hilar: 1 cyst, (4-a) paraesophageal-upper: 1 cyst, (4-b) paraesophageal-lower: 1 cyst, (5) CRAzV: 4 cysts, and (6) miscellaneous: 2 cysts. PMID- 8230924 TI - [Surgery for ventricular septal defect with aortic regurgitation]. AB - Twenty-two patients (mean age of 12.9 years) with ventricular septal defect (VSD) associated with aortic regurgitation (AR) were treated surgically. Sixteen patients had subpulmonic VSD and six had infracristal VSD. Fourteen patients with subpulmonic VSD underwent VSD closure alone. VSD was closed by pulling up the inferior rim to the pulmonary valve thus support the prolapse aortic cusp. AR improved in two and has remained stable in eleven for 0.5 to 12.7 years (mean 4.4 years). Aortic valve replacement (AVR) was necessary in an adult patient later. Plication of the aortic valve was effective in a young patient but ineffective in an adult patient. In infracristal VSD, AVR was necessary in two adult patients. In a young patient, AR improved by plication. Direct VSD closure ceased progression of AR in three patients. The long-term follow-up shows that VSD closure with our technique has been sufficient to arrest progression of AR in subpulmonic VSD. Early closure of VSD should be emphasized to prevent aortic valve prolapse and regurgitation. PMID- 8230925 TI - [Effect of extended thymectomy in myasthenia gravis of pure ocular type]. AB - The effect of extended thymectomy in myasthenia gravis of pure ocular type was investigated in 13 patients, who were divided into the surgical group of nine and the conservative group of four. In the surgical group, the onset was at 47 +/- 20 years old. The preoperative duration of symptoms was 4 +/- 6 years. The age at the time of operation was 51 +/- 16 years old. Three patients had thymomas, including one malignant thymoma. Acetylcholine receptor antibody titer was over normal range with 34.0 +/- 49.7 nmol/l. No postoperative crisis occurred. The palliation rate of the surgical group was more higher through the whole period than that of the conservative group, and increased with progress of the course. The remission occurred in the surgical group, but did not in the conservative group. It took 7 months to improve the symptom in the surgical group, while 18 months in the conservative group. We conclude extended thymectomy is reasonable for patients with myasthenia gravis of pure ocular type. PMID- 8230926 TI - [Buffering capacity of histidine in cardioplegia analyzed by myocardial production of lactate and alanine]. AB - The buffering capacity of histidine in cardioplegia was analyzed from a viewpoint of anaerobic glycolysis and its end-products (lactate and alanine) by the myocardium. Isolated rabbit hearts were subjected to 3 hours multidose cardioplegic arrest by histidine (195 mM)-buffered or control solution at 21 degrees C and 30 minutes reperfusion. Diastolic pressure (DP) and recovery of developed pressure (DevP%) were measured with an intracavitary balloon filled to a pre-ischemic DP of 8-10 mmHg. Lactate and alanine in tissue and effluent were measured after ischemia by HPLC. The buffered solution group showed lower DP and higher DevP%, production of more lactate and alanine with diffusion of larger part of them into effluent than control group. We conclude that histidine in cardioplegia stimulates production of anaerobic glycolysis-derived high energy phosphate-compounds not only by proton buffering but also by removal of detrimental end-products out of the cell. PMID- 8230927 TI - [Right thoracotomy to approach mitral or tricuspid valve in reoperation]. AB - Four patients underwent mitral valve re-replacement, and required tricuspid annuloplasty in two, through a right thoracotomy. There was no major postoperative morbidity and recovery was full and uneventful. The indications for selection of this approach were isolated mitral valve disease or combined tricuspid and mitral valve disease, severe adhesion between heart and sternum on CT scan, intact aortic valve and no severe reduction of respiratory function. We recommend right thoracotomy to approach atrioventricular valves in selected situations in patients for reoperations. PMID- 8230928 TI - [Two cases of rupture of intraaortic balloon]. AB - We reported two cases of rupture of intraaortic balloon (IAB). The most likely cause appears to be the rupture of IAB membrane against a calcific atherosclerotic aortic plaque. When the IAB ruptures by contact of calcific lesions in aorta, the leak detection of device of intraaortic balloon pumping is useless to diagnose this rupture. Therefore, the rupture of IAB through a pin hole is diagnosed by the transient presence of blood in the tube connecting the balloon. It is recommended that the balloon immediately should be removed at the transient presence of blood in the connecting tube. PMID- 8230929 TI - [Combined resection of the aortic wall in T4 lung cancer under the assistant devices]. AB - Radical operation for T4 lung cancer is technically difficult. We report here 3 cases of T4N0M0 lung cancer involving aorta or left subclavian artery which was successfully operated on. We could carry out combined aortic resection more safely and speedy under the assistant devices, such as Anthrontube and Biomedicus Centrifugal Pump. Even T4 lung cancer having aortic invasion with N0 disease can be operated on and be expected long-term survival using these assistant devices. PMID- 8230930 TI - [Treatment for cardiac rupture following acute myocardial infarction]. AB - We experienced thirty-three patients with cardiac free wall rupture following acute myocardial infarction (AMI). Twelve patients received reperfusion therapy. Surgical closure of cardiac rupture was performed in twelve patients under cardiopulmonary bypass (CPB). Two of these patients were placed on percutaneous cardiopulmonary support system (PCPS) immediately after the onset of blow out rupture. Cardiac rupture occurred in 19 patients within 3 days from the onset of AMI (early rupture) and in 14 patients 4 days after the onset (delayed rupture). All five patients with successful reperfusion were in the early rupture group, while there were no patients with successful reperfusion in the delayed rupture group. In 12 surgical patients, 7 patients had blow out rupture and 5 patients had oozing rupture. In 7 patients with blow out rupture, four patients could not be disconnected from CPB and 2 patients had cerebral death. One patient received PCPS support showed no mental disturbance after surgery, but he died of renal failure 6 days after surgery. Three out of 5 patients with oozing rupture survived. The interval from the onset of hypotension to establishment of cardiopulmonary support was 5 minutes and 60 minutes in two patients using PCPS. However it averaged 94 minutes in 5 patients using usual CPB. It is suggested that early reperfusion therapy may prevent delayed cardiac rupture following AMI, and surgical therapy using PCPS may be useful for the treatment of cardiac blow out rupture. PMID- 8230931 TI - [Surgical treatment for impending rupture of distal aortic arch aneurysm: a case report using "elephant trunk" prosthesis]. AB - A 75-year-old man with impending rupture of distal aortic arch aneurysm was operated on using the "elephant trunk" prosthesis. A long graft was anastomosed only to the proximal aortic stump and its distal free portion was inserted through the exit of the aneurysm into the descending aorta. Postoperative CT scan and MR image showed complete thrombosis formation in the dead space between the graft and the aneurysmal wall, which precluded necessity for further operation. However the graft was looped and kinked in the aneurysm. Furthermore the patient suffered from incomplete paraplegia, probably caused by the extremely long graft. In terms of promoting thrombosis formation in the aneurysmal inner space the longer prosthesis is feasible, but one should keep it in mind that an excessively long graft can lead to the kinking of the graft and paraplegia. PMID- 8230932 TI - [A case of coronary artery bypass grafting in a patient with active systemic lupus erythematosus]. AB - A 59-year-old female was admitted to our hospital because of chest pain. Coronary angiography revealed severe obstruction of the left coronary artery. The findings of hematology and renal biopsy were interpreted as showing active systemic lupus erythematosus (SLE). Coronary artery bypass grafting was performed before steroid therapy because of worsening cardiac function due to myocardial ischemia. Postoperative course had been satisfactory immediately after operation, but the patient showed pulmonary infection about two months after operation and died three months after surgery. In open heart surgery for a patient with active SLE, special care must be taken to the prevention and treatment for infection. PMID- 8230933 TI - [A case report of the rupture of the descending thoracic aorta due to blunt chest trauma]. AB - A 31-year-old man was injured in an automobile accident. He was suspected to have a rupture of the thoracic aorta because the widening of mediastinum was demonstrated on a chest X-ray film. Aortography (IV-DSA) was performed to confirm the diagnosis. Prosthetic graft interposition for the rupture of the aortic isthmus was accomplished with the aid of left heart bypass using the Biomedicus Centrifugal Pump with limited systemic heparinization (0.8 mg/kg). This adjunct appears to be useful to avoid the paraplegia and to reduce the blood loss. PMID- 8230934 TI - [Late malfunction of the Bjork-Shiley valve prosthesis due to Delrin disc defacement]. AB - A 55-year-old woman eighteen years after mitral valvular replacement with Delrin disc Bjork-Shiley valve prosthesis underwent a reoperation of prosthetic valve replacement for the prosthetic malfunction due to disc defacement. The patient suffered from faintness and vertigo at rest. An echocardiographic examination showed a moderate mitral insufficiency with a normal disc movement. Precise examination on the removed prosthesis revealed accelerated defacement of disc margin which made the ring-disc clearance up to 0.35 mm and strut-shaped groove formation on the inlet surface of the disc occluder. These findings suggested a pronouncedly earlier disc wear than predicted by Bjork and co-workers. We concluded, therefore, that a patient undergone a valve replacement with Delrin disc Bjork-Shiley valve should be examined periodically by echocardiography even though being without any symptoms. PMID- 8230935 TI - [A case report of cor triatriatum benefit of MRI for preoperative diagnosis and surgical method]. AB - Cor triatriatum is uncommon in all congenital heart diseases. It is a malformation resulting in a separation of the left atrium or right atrium into two chambers due to a congenitally abnormal diaphragm. We wish to present a case of cor triatriatum in which MRI was found most useful for preoperative diagnosis and surgical procedure. A 2-year-old girl was transferred to us for severe pulmonary congestion as shown on chest X-ray. Echocardiography showed abnormal diaphragm in the left atrium. MRI demonstrated clearly the relationship between left pulmonary vein and the abnormal diaphragm. Therefore we should preoperatively determine type I A according to the Lucas and Schmidt's classification. Cardiac catheterization showed moderate pulmonary hypertension and confirmed cor triatriatum. The resection of the abnormal diaphragm was performed under extracorporeal circulation with moderate hypothermia. The postoperative course was uneventful. MRI is a very useful non-invasive technique in making a diagnosis and in choosing the appropriate surgical procedure for cor triatriatum. PMID- 8230936 TI - [A case of congenital atrial flutter associated with atrial septal defect]. AB - We report the case of a 8-month-old boy with atrial septal defect associated with congenital atrial flutter. He was operated on for ASD successfully. Atrial flutter in infants has been reported to be uncommon and to have a poor prognosis when associated with underlying cardiac disease. Therefore, early surgical intervention may improve the prognosis. PMID- 8230937 TI - [A case of traumatic right diaphragmatic hernia]. AB - A 65-year-old female was sent to our hospital by ambulance car after being struck by a car. Chest X-ray showed a elevation of the right diaphragm. Computed tomography and liver scintigram revealed herniation of the liver into the right thorax. As the blood gas analysis was in normal range and cerebral contusion coexisted, she was observed without operation. During 5 year after the injury, the hernia was gradually progressive and dyspnea appeared finally. She was readmitted to the hospital and operated upon through thoraco-abdominal approach. There are no report of spontaneous cure of the hernia. When traumatic diaphragmatic hernia is diagnosed, an operation consistent with the patient's general condition should be performed at that time. PMID- 8230938 TI - [Torsion of the middle lobe after right upper lobectomy: a case report]. AB - Following right upper lobectomy for a pulmonary carcinoma, torsion of right middle lobe occurred. There have been few reports in the literature of pulmonary torsion after lobectomy. A 68-year-old man had a right upper lobectomy for pulmonary carcinoma. On the seventh day after right upper lobectomy, he got high fever. At the same time a roentgenogram of the chest showed complete opacification like ground-glass in the upper lung field. Gradually the opacification increased and hemosputum also appeared. Bronchoscopy revealed 90 degree torsion of the middle lobe. In the treatment with respirator (CV 2000), his clinical conditions improved. PMID- 8230939 TI - [An experience with surgical treatment for mucoepidermoid carcinoma of the lungs]. AB - Mucoepidermoid carcinoma (MEC) of the lungs is thought to arise in the bronchial glands. It is a tumor that rarely develops and it has a low grade of malignancy. In this paper, we describe one case of infiltrative MEC, which we were able to diagnose preoperatively. Surgery revealed a high grade malignancy which is reported here with a discussion based on the related literature. The patient was a 63-year-old male who was referred to our hospital by another physician due to a cough and left chest pain. A simple chest X-ray revealed a tumor shadow and a fascicular shadow on its periphery in the upper left lobe. Bronchoscopy disclosed complete circumferential stenosis at B1+2,3 and reddening from this region to the main bronchus, but it was impossible to directly confirm the tumor. Pulmonary arterography did not depict the left upper pulmonary vein, but obstruction due to a tumor of that vein was observed. Given the above findings, under a diagnosis of infiltrative MEC, a left total lobectomy accompanied by a combined left atriectomy was performed. Although most cases of MEC have a low grade malignancy, there have been some reported cases with a very high grade of malignancy. Therefore, evaluation of the progress of this type of carcinoma by preoperative diagnosis as well as radical excision appropriate to lung cancer are considered to be important. PMID- 8230940 TI - [Influence of thoracotomy and partial lung resection on pulmonary function: comparison with median sternotomy]. AB - The influence of thoracotomy on respiratory function was examined in 49 patients: 22 with intercostal thoracotomy without lung resection (ICT), 11 with partial lung resection (PLR) and 16 with median sternotomy (MST). Respiratory function (FVC, FEV1.0 and FEV1.0%) was measured preoperatively, in the early stage (14 days) and in the stable stage (after 3 months) postoperatively. In each group, the FVC and FEV1.0 were decreased to 70-80% of preoperative respiratory function in the early stage and rose to 90-95% in the stable stage. There were no significant differences among the groups. FEV1.0% did not vary in the different stages in any of the three groups. The ICT group was classified according to the type of skin incision (axillary incision (AI), anterolateral incision (ALI) and posterolateral incision (PLI)). FVC and FEV1.0 in the ALI subgroup were higher (p < 0.05) than in the PLI subgroup in the early stage. These results suggest that the selection of the skin incision is important for preserving respiratory function. PMID- 8230942 TI - Remarks to the American Medical Association. PMID- 8230941 TI - Antibiotic resistance of Escherichia coli and Salmonella from apparently healthy slaughtered cattle and pigs, and diseased animals in Zambia. AB - Escherichia (E.) coli and Salmonella (S.) choleraesuis (subsp. choleraesuis and subsp. salamae) from apparently healthy slaughtered cattle and pigs in 1989 in Zambia, were examined for antibiotic resistance and the presence of conjugative R plasmid. Salmonella strains from diseased animals (cattle, chickens, leopards, lions and warthogs) were similarly tested. The majority of the cattle had been nomadically kept in so-called "traditional farms" while all the pigs were from commercial farms. More pigs (39%; 41/105) harboured drug-resistant E. coli than cattle (6.7%; 7/105). Moreover, the number of drug-resistant E. coli was higher among strains from pigs (31.2%; 49/157) than cattle (4.2%; 7/167). For both cattle and pigs, drug resistance was more frequently observed against tetracycline, streptomycin, sulfadimethoxine and ampicillin than other antibiotics and the single resistance pattern occurred most frequency, especially among pig E. coli strains. Drug-resistant Salmonella was recorded in 3.6% (1/28) of strains from slaughtered cattle and 31.3% (10/32) of those from diseased animals. Drug-resistant E. coli from pigs and cattle carried R plasmid at high frequency. PMID- 8230943 TI - Treatment of human glioblastoma by specific immunotherapy. PMID- 8230944 TI - Orthotopic liver transplantation at KU Medical Center. PMID- 8230945 TI - The anesthetic management of liver transplantation. PMID- 8230946 TI - Magnesium for myocardial infarction. PMID- 8230947 TI - [Infant care by mothers in the patient's room in Ward 3B of Tampere University Hospital]. PMID- 8230948 TI - [Cost control and the quality of care]. PMID- 8230949 TI - [Practical education for nursing and current efforts for improving care in the Helsinki University Hospital's Maternity Clinic]. PMID- 8230950 TI - [Observations on the activities of midwives]. PMID- 8230951 TI - [Andreas was born in the midwifery kitchen]. PMID- 8230952 TI - [Greetings from Ethiopia]. PMID- 8230953 TI - [Family instruction is important]. PMID- 8230954 TI - [Ethics guidelines of the International Council of Midwives]. PMID- 8230955 TI - [The background of foreign patients with pulmonary tuberculosis and investigation of treatment results]. AB - This study focuses on the investigation of 35 foreign patients diagnosed as having pulmonary tuberculosis and treated at National Chiba-Higashi Hospital and the Chiba Anti-Tuberculosis Association between 1987 and 1991. All of the patients had lived in Japan less than five years. A large number of the patients were in their 20s. The chest X-rays of 24 cases (70%) revealed cavitary and sputum smear cultures for Mycobacterium tuberculosis were positive in 27 cases (77%). Many patients were in moderately advanced stages of the disease, becoming ill soon after their arrival in Japan, and being diagnosed symptomatically. Among the 35 cases, eight cases failed to finish treatment. Of those failing to finish, six were female and two were male. The results of sputum smear cultures of the patients failing to complete treatment revealed four cases that tested positive. Among those cases failing to complete treatment, four patients discharged themselves, three patients stopped during out-patient treatment, and one patient had to return to her home country because of a legal problem. A total of 14 patients were illegal immigrants: three of them failed to complete treatment because of their illegal status, and two of them due to unspecified reasons. PMID- 8230956 TI - [Serum and pleural SCC-antigen levels in patients with pulmonary tuberculosis]. AB - We measured the serum and pleural levels of squamous cell carcinoma related antigen (SCC-Ag) in patients with active pulmonary tuberculosis and studied the relationship between SCC-Ag and tuberculosis. Serum levels of SCC-Ag in 63 patients with newly diagnosed untreated tuberculosis were 1.76 +/- 2.16 ng/ml, significantly higher than in 118 healthy controls (0.67 +/- 0.52 ng/ml) or in 11 patients with old tuberculosis (0.92 +/- 0.38 ng/ml; p < 0.01). Pleural effusion levels of SCC-Ag in 41 patients with tuberculosis pleurisy were 4.5 +/- 3.4 ng/ml, significantly higher than in 54 patients with non-malignant, non tuberculous, pleurisy (2.3 +/- 1.7 ng/ml: p < 0.05). In patients who responded well to anti-tuberculous chemotherapy, serum levels of SCC-Ag decreased to the normal range (1.6 ng/ml) with clinical improvement, while in nonresponders they did not decrease. Isoelectolic focusing electrophoresis showed serum SCC-Ag to be composed of a neutral fraction as a single peak, which was corresponded to that of squamous cell carcinoma of the lung. The tuberculous lesions were stained with anti-SCC-Ag antibody in one case and weak SCC-Ag positive findings were observed in the intercellular spaces of the epidermoid cells in the area without necrosis. These results raise the possibility that SCC-Ag is produced in or released from not only squamous cell carcinoma of the lung but also pulmonary tuberculous lesions. PMID- 8230957 TI - [Clinical features of primary and secondary atypical mycobacteriosis]. AB - A clinical investigation was carried out in 39 patients with atypical mycobacteriosis (AM). The patients at our hospital diagnosed as having AM during an 11-year period from 1981 to 1991 were reviewed in this study. The incidence of AM among all of mycobacterial infections has been increasing since 1988. The atypical mycobacteria detected included M. avium complex (MAC) in 79%, M. kansasii in 26%, and other organisms in 5%. Patients who had primary infection without underlying respiratory disease were compared with those who had secondary infection. In secondary infection, MAC was detected in the sputum of a high percentage of patients and was positive for more than 6 months despite therapy. Gamma globulin levels were also increased in patients with secondary infection. Cell mediated immunity was examined by the DNCB reaction and the detection of CD4 and CD8- positive T lymphocytes. The DNCB skin reaction was positive in 6 out of 26 patients with secondary infection. The percentage of CD8- positive T lymphocytes was higher in patients with secondary disease. Fischer's ratio (indicating the balance of plasma amino acids) was also examined as an index of the nutritional status. It was significantly lower in secondary infection compared with primary infection. Moreover, secondary infection was associated with a lower positive rate for the DNCB skin reaction and a lower Fischer's ratio when compared with pulmonary tuberculosis. Thus impairment of cell-mediated immunity, malnutrition, and a poor prognosis were significantly more common in secondary infection compared with primary infection. PMID- 8230958 TI - [A case of tuberculosis of the thyroid gland]. AB - A 49-year-old woman was admitted to our university hospital for examination of a neck mass. Computed tomography and ultrasonography revealed a nodule in her thyroid gland and a swollen cervical lymph node. They were removed surgically; a subsequent histological study revealed an epithelioid cell granuloma. A diagnosis of localized tuberculosis of the thyroid gland and a cervical lymph node was thus confirmed. The data of the present case were discussed with reference to the characteristics of thyroid tuberculosis described in the literature. PMID- 8230959 TI - [A dementia occurred during therapy for multi-drug resistant pulmonary tuberculosis]. AB - A 47-year-old woman was thirdly admitted to our hospital for therapy to multi drug resistant tuberculosis of the lung in June 88. Although she was treated with TH, PAS and EVM, M. tuberculosis were positive constantly in her sputum. She complained of sleep disturbance, irritability, headache on September 89, she was diagnosed reactive depression. But her symptoms were progressive, low level of intelligence test in November, apatic in December and became spastic paralysis and vegetable state at next year. She was died of pneumonia in November 90. Her autopsy findings showed no brain atrophy nor meningitis. Microscopically, central chromatolysis were showed in Betz cells and anterior horn cells, these findings sometimes suspected for Peragulla, but we could not obtained definite pathological diagnosis, because we could not have been obtained findings indicating for Peragulla. Finally, we reported the clinical course and autopsy findings of dementia occurred during therapy for multi-drug resistant pulmonary tuberculosis. PMID- 8230960 TI - [The 68th annual meeting symposium. III. Changes in the disease picture of pulmonary tuberculosis]. AB - In the symposium on 'Changes in the disease picture of pulmonary tuberculosis', the themes mentioned above were discussed. Dr. Ohmori analyzed the present state of the epidemiology of tuberculosis in Japan. The stagnation in the decline of incidence rate of tuberculosis has been observed from the end of the 1970's. It is remarkable in the population aged 15-29 years. She noted two factors as the reason of such stagnation. Firstly, the number of tuberculosis cases increases in the high-aged subjects who were infected with tubercle bacilli in the past, as the population with high-age is expanding. Secondly, sporadic mass outbreak of tuberculosis is found often in the young subjects who have not been infected with tubercle bacilli. Dr. Miyazaki reported that 57 cases (8.5%) were classified as lower lung tuberculosis in 672 new cases with pulmonary tuberculosis during the last ten years. He noted the usefulness of fiberoptic bronchoscopy for diagnosis to lower lung field tuberculosis. Dr. Nagata analyzed the present state of mycobacterial disease in 5041 autopsy cases. The rate of cases with active mycobacteriosis was about 1% during the last 5 years. A large proportion of cases with fatal mycobacteriosis was received corticosteroid therapy. Antemortem diagnosis of mycobacteriosis was made in 25.9-20.0% of cases with fatal mycobacteriosis. Dr. Yamagishi showed the data of pulmonary tuberculosis in compromised hosts in a National Hospital. There were 82 cases associated with various type of cancer (4.2%) and 21 cases treated with corticosteroids (1.1%) among 1954 cases with pulmonary tuberculosis for the last 6 years.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230961 TI - [The factors related to the stagnation in the decline of tuberculosis incidence in Japan]. AB - The mortality from tuberculosis in Japan had extremely decreased from the end of the 1940's to the beginning of the 1950's, due to the end of chaos after World War II and also due to the introduction of antituberculosis drugs. The rapid decline of mortality usually leads to the reduction in the infection risk of tuberculosis in the general population. Such a drastic change in the history of tuberculosis have been dividing general population into two groups, i.e. those who were born during the rapid spread of infection with tubercle bacilli and others who were born after. With the passage of time, the limiting age between those two groups reached 40-50 years old as of 1980. The elderly people who were infected with tubercle bacilli in the remote past, have a high risk of development of tuberculosis by endogenous breakdown. In addition, the population of the aged in Japan has been expanding very fast as compared to that in European countries. Owing to such a change of historical and demographical background, the proportion of the elderly cases developed to tuberculosis have increased considerably and it played a major role in a small decrease of tuberculosis incidence rates since 1980. On the other hand, a generation gap on the prevalence of tuberculosis infection caused the recent smallest reduction rate of incidence among young adults. As the majority of young people have not been infected with tubercle bacilli, since 1980, the micro-epidemic among adolescent and young adults have been reported often.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230962 TI - [Lower lung field tuberculosis]. AB - Lower lung field tuberculosis has been considered as rare disease, but it was observed about ten percent in the literature. It seemed that the important problems in lower lung field tuberculosis are delay and difficulty of diagnosis. The materials for this report was accumulated over a period of 10 years, from January 1983 to December 1992 at the National Sanatorium Fukuoka-Higashi Hospital. The criteria for lower lung field tuberculosis are: (1) disease confined to lower lobe except for S6a, middle lobe and lingular segment, (2) bacteriologically and/or histologically proved diagnosis of tuberculosis, (3) clinically considered as tuberculosis for which improved by only anti-tuberculous drugs. During this period, 672 new patients suffering from active pulmonary tuberculosis were admitted to the sanatorium. Of the total of 672 patients, 57 patients were selected as the criteria for lower lung field tuberculosis. The incidence of them was 8.5 percent of the total new admissions. Of these 57 patients, 30 were females and 27 males. The mean age of these 57 patients was 59.4 years ranging between 19 and 86 years. The incidence of them obtained through ten years did not show any tendency to increase or decrease. The respiratory and general symtoms most commonly observed were cough and chest pain. The average duration of used for diagnosis of the lower lung field tuberculosis over 65 years old was 71.7 days and it seemed to show the delay and difficulty of diagnosis. PMID- 8230963 TI - [Pulmonary tuberculosis in compromised hosts in pathological view--the present state of mycobacterial disease in autopsy cases in two university hospital]. AB - To clarify the present state of mycobacterial disease in autopsy cases, we reviewed the case records and autopsy data of 3848 cases who died in Kyushu University Hospital between 1976 and 1990, and 1193 cases died in University of Occupational and Environmental Health between 1981 and 1990. The incidence of mycobacterial disease has significantly decreased during the period examined, and that of active disease was about 1% between 1986 and 1990. Significantly more cases of fatal mycobacteriosis were received with corticosteroid than those of active but not fatal mycobacteriosis. Antemortem diagnosis of mycobacteriosis was made in 25.9% and 20% of fatal cases in each University Hospital. The possibility of mycobacteriosis was not suspected at all in some cases of fatal mycobacteriosis. Regardless of the suspicion of mycobacterial disease, the diagnosis was not made in other fatal cases, because of the negative bacteriology. PMID- 8230964 TI - [Pulmonary tuberculosis in the compromised host at the point of clinical view]. AB - The rate of the compromised host tend to increase in the pulmonary tuberculosis due to long term use of immunosuppressive drugs, such as anti-cancer drugs and corticosteroids. On the one hand, the incidence of tuberculosis has reduced in Japan, so, many doctors consider tuberculosis as a disease of the past, and doctor's delay is the serious problem. We investigated pulmonary tuberculosis in the compromised host. 1) Chiba University Hospital. For five years, the 77 cases were diagnosed as pulmonary tuberculosis, for their sputum examination were positive. The 54 cases didn't have other diseases, the 17 cases had various type of cancer, and only one case was administrated with corticosteroid. 2) National Chiba-Higashi Hospital. For 6 years, the 82 cases had various type of cancer and the 21 cases were administrated with corticosteroids, in the 1954 cases of pulmonary tuberculosis patients in our hospital. In the cases having cancer, the 18 cases had lung cancer, the 36 cases had cancer of the digestive organs. In the cases of having lung cancer, the 12 cases died of cancer and no cases died of pulmonary tuberculosis. In the cases of having cancer of the digestive organs, the 17 cases died of cancer and the 3 cases died of pulmonary tuberculosis. In the cases administrated with corticosteroids, the 17 cases had been treated for collagen disease. In these cases, many patients were progressive state on chest roentogenograms. When their onset of pulmonary tuberculosis, the 12 cases were administrated with low dose of corticosteroids. In all cases, chemoprophylaxis had not been done.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230965 TI - [Tuberculosis in foreigners from Asian and African countries]. AB - 121 patients with active tuberculosis, who recently arrived from Asian and African countries and were admitted to our hospital between 1986 and 1991 for treatment, were clinically reviewed and compared with 177 Japanese patients. The notable features of tuberculosis in these foreigners were relatively short intervals from arrival to onset, in younger population, higher initial antituberculous drug resistance, longer conversion interval, poor compliance with treatment and more frequent dropout cases. Tuberculous pleuritis were happened more frequently in Japanese group, possibly indicating initial infection. It is supposed that many of foreign group were infected in origin countries and reactivated before or after arrival, and it may account for higher initial drug resistance. Only 73% of foreign group were completed standard chemotherapy, where as 88% of Japanese group. Main causes of discontinuation were returning to origin counties and poor compliance. In addition, increasing of illegal stay will need administrative programs for control. PMID- 8230966 TI - Violence is becoming more visible! PMID- 8230967 TI - Sexual assault: theories and treatment of the offender. PMID- 8230968 TI - Assessing and addressing violence in the acute care setting. PMID- 8230969 TI - Guns in roses. PMID- 8230970 TI - [Immunologic aspects of allogenic valve implantation]. AB - Immunological response against myocardial antigens was assessed in 12 patients (pts) (9 males, 3 females, aged 24-62 years) with aortic homografts (AH) and 36 pts (29 males, 7 females, aged 26-61 years) with aortic valve prostheses (AVP). In all pts a titre of anti-myocardial antibodies, a concentration of free circulating immune complexes and leucocytes migration inhibition test with myocardial antigen were assessed before operation and 1, 12, 24 months after operation. In 20 pts with AH immunological humoral response against HLA antigens was evaluated. After the operation pathological indexes of immunological response against myocardial antigens were observed both in pts with AH as in pts with AVP. The mean titre of anti-myocardial antibodies before operation was in both groups between 1:4 and 1:8, and 1 month after operation between 1:32 and 1:64. In 12 and 24 months after operation the mean titre in AH and AVP group was respectively 1:8 and 1:4. Before operation pathological concentration of immune complexes (> 0.084 mg/ml) was observed in 17% AH and 11% AVP pts. Pathological elevation of immune complexes 1, 12 and 24 months after operation was observed respectively in AH and AVP group in 58%, 42%, 33% and 67%, 50%, 42%. Leucocyte migration inhibition index was pathological (< 0.8) before operation in 58% in both groups, and 1, 12, 24 months after operation was pathological in AH and AVP group respectively in 67%, 17%, 17% and 61%, 33%, 31%. In most pts of both groups immunological response against myocardial antigens was transient and was terminated up to 12-24 months after operation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230971 TI - [Localization of myocardial infarction with the help of EKG and two-dimensional echocardiography. Controversies]. AB - A multicentre study of the localization of myocardial infarction (MI) using ECG and 2-dimensional echocardiography (Echo-2D) was performed on the 21st day of the onset of MI. The study population consisted of 650 pts (mean age 55.0 years), 553 males and 97 females. The purpose of the study was: 1) to compare the site of MI as diagnosed by ECG and Echo-2D, 2) to determine the controversies in the diagnosis between these two methods. Consistent results of both methods were obtained in 408 pts (62.8%) of the group. In 61 pts (9.4%) the diagnosis of MI by ECG and Echo-2D was undefined. In 181 pts (27.8%) the inconsistencies of ECG and Echo-2D evaluations were demonstrated; in 106 pts ECG changes were undefined, but evident Echo-2D changes were found; on the contrary, in 51 pts MI diagnosed by ECG was not confirmed by Echo-2D. In 24 pts entirely inconsistent results were shown. 209 pts (32%) with myocardial contractility disorders in the apical region of the heart diagnosed by Echo-2D showed different MI localisation as determined by ECG: 147 pts had anterior or antero-lateral MI, 28 pts--postero-inferior MI, 12 pts--apical MI and 22 pts--another one. By these findings it has been shown that ECG and Echo-2D are compatible methods but not replaceable ones. PMID- 8230972 TI - [Atrial fibrillation with chronic obstructive pulmonary disease]. AB - We compared 8 patients with COPD and fixed atrial fibrillation (group I) and 46 patients with COPD and sinus rhythm in ecg (group II). None of the patients had valvular heart disease, arterial hypertension or clinical signs of coronary artery disease. The studied groups did not differ while comparing their gas measurements, spirometry, pulmonary hypertension and right ventricular diameter (as measured in echo study). Patients with fixed atrial fibrillation had larger circuit and area of right atrium (p = 0.001), left atrial and left ventricular diameter (p = 0.001) as well as lower LV function (expressed by FS%). In conclusion, enlargement of both atria may be considered as a cause for fixed atrial fibrillation in patients with COPD. However symptomless coronary artery disease rather then COPD may be the reason for this arrhythmia. PMID- 8230973 TI - [Permanent cardiac pacing in children. Personal experience]. AB - Pacemaker implantation was performed in 26 children aged from 5 months to 15 years (mean 4.6). The indications for pacemaker implantation were: sick sinus syndrome in 4 patients, III degrees idiopathic A-V block in 2 both with congenital heart disease and III degrees post-op. A-V block in 20 patients. 34 pacemakers produced by seven different companies were implanted. Epicardial electrode was used in all but one patient in whom endocardial electrode was introduced. Demand for impulse voltage was 2.4-5.2 V (mean 4.4 V), impulse duration was from 0.25 ms to 1.65 ms (mean 0.62 ms). Three out of 26 pts. (11.5%) died (1-intraoperative bleeding, 1-ventricular rhythm disturbances, 1-sudden death). In seven children first exchange of the pacemaker (one for mechanic damage and 6 for exhausted batteries) was performed mean 4.3 years after previous implantation. In two children second battery exchange was necessary mean 7.5 years after previous (both exhausted batteries). In five children a damaged epicardial electrodes were exchanged (21%). PMID- 8230974 TI - [Surgical closure of atrial septal defect--when is it too late?]. AB - We describe a 41 year-old patient with atrial septal defect combined with mitral stenosis, severe pulmonary hypertension and tricuspid regurgitation. Despite high pulmonary vascular resistance, surgical intervention was decided on. Follow up proved to be successful. The role of echocardiography and hemodynamics data to establish a preoperative diagnosis is discussed. PMID- 8230975 TI - [Pseudoaneurysm of both superficial femoral arteries complicating coronary angiography: case report]. AB - A 58 years-old patient with aortic valve disease was admitted to our hospital for coronarography. Coronarography by transfemoral route bilaterally failed and eventually the coronary arteries were visualised by the transaxillary route. After the study bilateral pseudoaneurysms of the superficial femoral arteries were diagnosed clinically and confirmed by color-Doppler and angiographic studies. Surgical therapy was successful. Causes, nature, diagnosis and surgical as well as nonsurgical therapy of the femoral artery pseudoaneurysms complicating percutaneous transfemoral cardiac catheterization are discussed. PMID- 8230976 TI - [Electrophysiologic examination in surgical treatment of patients with Wolff Parkinson-White syndrome]. PMID- 8230977 TI - [Stimulators of the heart with adapted frequency of impulses]. PMID- 8230978 TI - [Intravenous ablation of the atrio-ventricular junction in patients with supraventricular tachyarrhythmias]. AB - Since the first successful therapeutic DC ablation of the AV junction in 1986, we have treated 20 symptomatic patients with drug-refractory supraventricular tachyarrhythmias (average of 6 antiarrhythmic drugs prior to the ablation attempt). The primary rhythm disturbances necessitating ablation were: AV nodal reentrant tachycardia (50% of pts), atrial flutter or fibrillation, with an uncontrolled rapid ventricular response (40%), atrioventricular reentrant tachycardia using an accessory pathway (20%), atrial tachycardia (10%), and junctional reciprocating tachycardia (5%). Percutaneous catheter ablation of the AV junction was made by Gallagher's method. The USCI 4-polar catheter (7F) was used in 40% of pts, and bipolar Cordis catheter (5F) in the remaining 60%. 70% of pts received either one or two shocks, usually of 200 or 300 J during one session. Another 25% received stored cumulative energy from 800 to 1200 J (in two sessions), and one patient--1800 J (during three sessions). In 85% of pts, the immediate post-ablation conduction was third-degree AV block with the escape pacemaker, ranging from 20 to 50 beats/min., which was infra-His in 57%, and supra-His in 43% of pts. In 15% of pts were either first-degree AV block (10%) or normal AV conduction (5%). A His bundle deflection more than 0.2 mV was predictive of successful production of third-degree AV block. Except a mild and transient increase of indicating enzymes (CPK and CPK-MB) we did not observe any other serious complications directly related to the ablalation procedure. Follow up study included 19 pts (time range from 2 to 56 months, mean 28).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8230979 TI - [Echocardiographic evaluation of funnel chest after surgical intervention]. AB - An analysis of echocardiographic examination in 22 patients operated for funnel chest is presented. Patients were divided into two groups: group I--isolated anomaly, and group II--patients with coexistent other postural deformations. During the operation, rib's cartilages in the parasternal region were excised, sternum was cut transversally above the deformation and longitudinally across it, and than anterior thoracic wall was stabilized in the proper position with Kirschner's wires. Before and 6 months after surgery echocardiographic examinations were performed with systolic, diastolic and ejection volume indices calculation. marked haemodynamic improvement was noted with the increase of diastolic and ejection volume of both heart ventricles. Improvement was more evident in patients with severe deformations and with coexistent other postural anomalies. PMID- 8230980 TI - [Doxazosin (alpha 1-adrenoreceptor antagonist) used in primary hypertension and it's beneficial effect on lipid metabolism]. AB - The aim of the study was to investigate the effect of a new selective alpha 1 adrenoreceptor blocker doxazosin on blood pressure, serum lipids and lipoproteins in patients with essential hypertension. The study was done in 32 out-patients with mild-to-moderate hypertension (22 men and 10 women, mean age 45.6 +/- 10.1). After 2-week placebo period the patients were given doxazosin in increasing doses from 1 to 8 mg daily for 6 to 14 weeks (mean daily dose 2.24 +/- 1.6 mg). Twenty nine patients completed the study. The active treatment caused highly significant drop in systolic and diastolic blood pressure both in supine and standing positions. No orthostatic hypertension was noted. There was also a statistically significant decrease in serum total cholesterol, VLDL-cholesterol, and triglyceride levels and increase in the positive prognostic lipid indicators. HDL3-cholesterol and apolipoprotein AI levels as well as HDL/total cholesterol ratio. Accordingly, the statistically significant decrease of the so called atherogenic index was noted. The drug was well tolerated and only one patient dropped from the study because of side effects. The authors conclude that doxazosin appears to be an effective and well tolerated antihypertensive drug with a favorable effect on lipid metabolism. It may be particularly useful in hypertensive patients with coexisting lipid abnormalities. PMID- 8230981 TI - [Ventricular fibrillation in a patient with mitral valve prolapse syndrome]. AB - A patient with mitral valve prolapse who had cardiac arrest as a result of ventricular fibrillation is described. After two weeks of unconsciousness all brain functions gradually improved and full intellectual efficiency was regained. Exact study of the case did not reveal any other possible reason for ventricular fibrillation but mitral valve prolapse syndrome. PMID- 8230982 TI - [Syncope; causes, significance, diagnostic progress]. PMID- 8230983 TI - [Magnesium in cardiology. Instant success of an undervalued ion]. PMID- 8230984 TI - [Electrophysiologic effects of magnesium]. PMID- 8230985 TI - [Head-up tilt table test in patients with syncope of unexplained origin]. AB - We studied 41 patients (21 m, 20 f) aged 22-70 years (mean 51) with syncope of unexplained origin using 60 degrees head-up tilt test (TT). Continuous ecg recording and direct arterial blood pressure were monitored. In case of positive response (e.g. syncope) in first examination next TT after administration of atropine (0.02 mg/kg) was performed. In first TT syncope was evoked in 19 patients (46%). Mean time from onset of tilt to syncope (TOS) was 15 +/- 13 min. Atropine administration protected against syncope during second TT in 7 patients, prolonged TOS in 4 (from 12 +/- 7 to 34 +/- 4 min) and shorted TOS (from 26 + 17 to 16 + 14 min) in the next 8 patients. We distinguished on that base: in 7 patients cardioinhibitory form, in 4 mixed form and in the next 8 patients vasodepressor form of vasovagal syndrome. We used in treatment: a) implantation of DDD or VVI pacemaker in 7 patients, b) clonidine in 8 patients, c) propranolol in 2 patients, d) derivates of atropine in 2 patients. During 1 year follow-up 14 patients are free of syncope and in the next 5 syncope occurred rarely. CONCLUSIONS: 1. Head up tilt test is very useful in diagnosis of syncope of unknown origin. 2. Second tilt after administration of atropine in patients with syncopal response in the first test helps to differ the type of vasovagal syndrome and to improve its treatment. PMID- 8230986 TI - [Carotid sinus massage in diagnosing syncope of unknown origin]. AB - Carotid sinus massage (CSM) was performed in 44 patients (23 female, 21 male, mean age 52 years) with recurrent episodes of unexplained syncope and without any organic heart disease. The protocol of examination consisted of 5 s CSM performed unilaterally in supine, upright 60 degrees positions, after administration of propranolol (i.v. 0.1 mg/kg) and during autonomic inhibition (atropine i.v. 0.02 mg/kg after propranolol). The diagnosis of hypersensitive carotid sinus syndrome (HCSS) was established in case of pathologic response to CSM in supine and upright 60 degrees positions. Positive results of CSM was obtained in 15 (34%) examined patients. Cardioinhibitory form of HCSS was recognized in 9 and mixed form in 6 cases. Pathologically augmented carotid sinus reflex was provoked in 8 cases by massage of the right, in 4 cases of the left and in 3 both carotid sinuses. Maximal time of electrical asystole after CSM was 6300 ms while maximal decreased od systolic blood pressure was 70 mmHg. Intravenous administration of propranolol caused decrease of systolic blood pressure approximately 43 +/- 15 mmHg whereas the cardioinhibitory effect of CSM after this drug significantly was increased only in patients with a positive response to CSM in the supine or upright 60 degrees positions. Autonomic inhibition significantly decreased the cardioinhibitory effect of CSM without significant influence on its vasodepressive component. PMID- 8230987 TI - [Prehospital death rate in acute myocardial infarction]. AB - The aim of the study was the analysis of prehospital death rate caused by AMI depending on sex, age and place of habitation in an agricultural-industrial region. Prehospital death rate resulting from AMI reached 34.6% (32.8% men and 39.3% women). 30.4% of patients died before obtaining medical help. The incidence of deaths increased with the more advanced age (until 70 was between 20-25% whilst, over 70 as high as 62-94%). PMID- 8230988 TI - [Magnesium protects myocardium during electric cardioversion but does not increase effectiveness]. AB - Transient elevation of serum enzyme level and abnormalities of repolarization following electrical cardioversion (EC) are usually observed. We aimed to alleviate these alterations by pretreatment with magnesium sulfate (MgSO4). The study group consists of 100 patients (aged 19-65 years) who had EC (300 W/sec) for chronic atrial fibrillation after mitral and/or aortic valve surgery. They were randomly divided into two sub groups: 50 patients in whom MgSO4 was administered intravenously (dose 100 m/kg b.w.) and 50 control subjects. Total serum magnesium level and CKMB level were recorded before and 8 hours after EC. Recovery to sinus rhythm, QTc interval, segment shifts were recorded immediately before and after EC. Results (mean +/- SE): [table: see text] Our results suggest that MgSO4 administered before EC has a significant cardioprotective effect and reduces abnormalities of repolarization. PMID- 8230989 TI - [Invasive treatment of recent myocardial infarction. Evaluation of left ventricular function]. AB - Hemodynamic parameters of left ventricular function were assessed in 179 patients with acute myocardial infarction, who were treated with intracoronary thrombolysis during first six hours after the onset of chest pain. Some of them were subjected to coronary angioplasty. Patency of the infarct related artery was controlled between the 2nd and 3rd week after acute myocardial infarction concomitant with evaluation of EF and LVEDP. Reperfusion of the infarct related artery was obtained in 121 patients (67.6%) and also significant increase of EF was observed in those patients. EF rose in patients with a patent coronary artery after 2-3 weeks (not significant) in contrast to patients with obstructed coronary artery in whom there was a fall in EF. The rise of EF was more pronounced in patients with reperfusion reached in 3 hours after the onset of infarct pain and without coronary angioplasty. It was shown that LVEDP is not usefull in estimation of left ventricular function. PMID- 8230990 TI - [Level of endothelin increases in chronic congestive heart failure]. AB - Endothelin is a family of potent vasoconstrictor peptides of vascular endothelial origin. Although it has been proposed that the vasoconstrictor effects of endothelin are produced at the local vascular level, the current study was performed to test the hypothesis that plasma endothelin concentrations are elevated in patients with chronic congestive heart failure. Plasma levels of endothelin were measured with sensitive and specific radioimmunoassay. Venous blood samples were obtained after 60 minutes of supine rest and serially during 40 minutes in the upright position. It was found that patients with congestive heart failure had markedly higher basal levels of circulating endothelin than subjects with coronary heart disease without heart failure and there was no further increase on postural change. Increase in plasma endothelin during upright position in subjects with coronary heart disease without heart failure, preceded the increases in circulating levels of the other vasoconstrictor mediators, consistent with the role of endothelin in neurohumoral compensation for hemodynamic stress. PMID- 8230991 TI - [Results and complications of PTCA]. AB - Between January 1987 and June 1992 we performed 189 PTCA procedures in a group of 153 patients (127 men, 26 women) aged 28 to 69 years, mean 52.3. 203 coronary vessels were dilated. In 136 pts one vessel was dilated, one vessel with two narrowings were dilated in 16 pts and in 17 pts two-vessel PTCA was performed. Due to restenosis, PTCA was repeated, once in 26 pts, twice in 3 pts and three times in 1 patient. Effective dilatation was achieved in 81.8% of cases and in this number, circumflex and marginal branch of left coronary artery were successfully dilated in 90.7%, left anterior descending in 84.6%, right coronary artery in 84.2% of cases. Attempts to recanalize these vessels were effective in 52%. There were no lethal complications of PTCA, acute occlusion of artery occurred in 5.9% of dilated vessels. Emergency coronary bypass had to be performed in 5.2% pts due to acute artery occlusion. PTCA supported by surgical stand-by appears to be a safe procedure with high direct effectiveness. PMID- 8230992 TI - [Tendon of Todaro in the human heart]. AB - The material consists of 50 human hearts without either pathological changes or congenital malformations. Macroscopic and microscopic methods were applied. Histological specimens were stained alternatively with hematoxylin-eosin and according to van Giesson and Masson. In each heart the tendon of Todaro was dissected from its portion situated between the inferior vena cava and coronary sinus up to central part of the membranous septum (Fig. 1). The hearts were divided into 3 groups according to age: fetuses (20 hearts), infants (15) and adults (15) (Table I). In all hearts of fetuses and infants the tendon of Todaro was found to be a well-developed cylindrical structure covered with endocardium. In histologic specimens the tendon was s solid structure well-separated from other tissues (Fig. 3, Fig. 4). In hearts of adults (17-45 years old) the tendon of Todaro was less evident. However, in histological specimens it was present as a compact connective tissue band (Fig. 2). In hearts of subjects older than 50, the endocardium of this area was not elevated and not distinguishable even by palpation. The connective tissue band was formed by rather dissipated fibres, poorly separated from surrounding structures. As a result of our study one may conclude that the tendon of Todaro is present in each human heart. It is well developed in hearts of fetuses and infants. Later it diminishes gradually becoming almost inconspicuous in old subjects. PMID- 8230993 TI - [Hormones regulating volume and electrolytes of sweat in patients with essential hypertension. Effect of thermal dehydration and treatment with captopril]. AB - The present study aimed to answer the following questions: 1. do secretion of volume related hormones in patients with EH pre and post treatment with captopril differ from normotensive subjects if examined in thermal dehydration conditions; 2. is the electrolyte composition of thermal sweat related to the plasma profile of volume related hormones? and 3. does treatment by captopril influence sweat electrolytes in EH patients. In 16 patients with EH and in 20 healthy subjects a thermal dehydration test was performed. In patients with EH this test was done twice: before treatment and after 6 weeks of captopril therapy. In all subjects plasma renin activity (PRA), aldosterone (Ald) AVP and ANP were measured before and after thermal dehydration. In sweat samples collected after 15' and 45' of thermal dehydration the concentration of Na, K and Cl was assessed. In hypertensive patients before captopril treatment significantly higher values of PRA, ALD and ANP were found, while sweat concentrations of Na and Cl were significantly lower than in controls. After captopril treatment sweat electrolytes concentrations showed a tendency to normalize. No significant correlation was found between the plasma hormonal profile and sweat Na, K and Cl concentrations respectively both in controls and patients with EH pretreatment. A significant positive correlation was noticed only in hypertensive patients post treatment between plasma aldosterone and sweat Na and Cl concentration respectively. Results obtained in this study show, that volume related hormones (Ald, AVP, ANP) do not seem to influence markedly the electrolyte composition of thermal sweat both in healthy subjects and in hypertensive patients. PMID- 8230994 TI - [Treatment of 3 patients with arrhythmias using "radiofrequency" ablation]. AB - In 3 patients radiofrequency (RF) ablation of AV node (n = 2) and left sided accessory pathway (n = 1) was performed because of intractable tachyarrhythmias. RF ablation was made in the first patient (ablation of AV node) using 135 Watts during 72 sec., in the second patient (AV node ablation) 331 Watts during 185 sec., and in the last patient 883 Watts during 695 sec. In the last patient ablation of accessory pathway was unsuccessful and the patient has been successfully operated. AV conduction was interrupted in 2 cases. No complications were noted. General anesthesia was not required. All the patients were discharged in a good state. PMID- 8230995 TI - [Electrical ablation with high frequency (radiofrequency) in patients with recurrent atrioventricular nodal reentrant tachycardia--introductory information]. AB - In 10 patients (pts) with atrioventricular nodal reentrant tachycardia type slow/fast (AVNRT) refractory to the antiarrhythmic treatment, radiofrequency current catheter ablation was performed. Adult pts (mean age 31 years) suffered from paroxysms of AVNRT for 2 to 18 years (mean 8 years). Episodes of arrhythmia recurred from 2-3 times a week to 1 for several months during antiarrhythmic therapy. The mean rate of the tachycardia was 243/min. 3 pts had syncope and 5 fainted during the tachycardia. In 2 females mitral valve prolapse was diagnosed. Programmed stimulation of the right atrium on the basic rate 130/min revealed ERP of the slow pathway mean 238 ms and ERP of the fast one mean 346 ms. The tachycardia has been induced with right atrium stimulation--most often programmed or burst stimulation--200 and 220/min, sometimes with incremental pacing. Successful radiofrequency energy ablation was performed in all 10 pts. Average number of the impulses for one pt was 7.7 (from 1 to 18 impulses). The total average time of the ablation was 216 s. The temperature of effective ablation was average 65 centigrade. The effect of the ablation was evaluated during repeated electrophysiological study before and after intravenous injection of atropine. Ablation was considered successful when no tachycardia was induced. P-R interval did not change and no episodes of AVNRT was observed in the 1-3 months follow-up. No complications were observed during the ablation and after the procedure, especially no thromboembolism was noticed. The echocardiographic evaluation revealed no damage to the heart structure. The pts receive no antiarrhythmic treatment and they are systematically controlled. PMID- 8230996 TI - [Electric ablation in St. George's Hospital]. PMID- 8230997 TI - [Diagnostic difficulties in a 32-year old patient with cardiac tamponade]. AB - At present the most frequent cause of cardiac tamponade is neoplasma. A case of a 32-year-old male with recurrent cardiac tamponade caused by pericardium mesothelioma is described. Despite advanced neoplastic process in the pericardium, pleura and mediastinum, neither radiologic examinations of the chest, echocardiography, nor repeated cytologic examination of the pericardial exudate, could establish the etiology of the tamponade. Only after a pericardial window had been performed was it possible to: 1) establish the diagnosis and introduce causal treatment; 2) prevent recurrence of the tamponade; 3) perform cytoreduction of the tumor. It seems that in cases of chronic exudative pericarditis of unknown cause it is proper to perform an early pericardial window as a diagnostic and therapeutic procedure. PMID- 8230998 TI - [Aortic-left ventricular tunnel--description of two cases]. AB - This is a very rare congenital anomaly, which clinically imitates aortic insufficiency and therefore should be kept in mind by the cardiologist. Only an early precise diagnosis and proper surgical treatment will result in a good long term prognosis. We have diagnosed and treated two patients, four and five years old, referred to us with a diagnosis of aortic insufficiency. Our final assessment was based on an obligatory cine-angio study, since a certain percentage of patients with this anomaly have concomitant lesions in the cardio vascular system, which should be precisely detected. A brief differential diagnosis and discussion concerning different surgical techniques are presented. In our two cases obliteration of the tunnel using a Dacron patch inserted vertically in the tunnel with closure of both ostia was performed. This technique seems to be superior compared to the previous one in terms of preventing the progressive destruction of the aortic valve. All the authors agree that early surgical treatment is indicated before the left ventricle becomes dilated and the aortic valve significantly damaged. Two years post surgery our patients are in very good condition with: no signs and heart failure, a gradual diminution of the left ventricular cavity and satisfactory function of the aortic valve. PMID- 8230999 TI - [Coexistence of WPW syndrome, coronary disease and segmental disorders of myocardial contraction. Diagnostic difficulties. Description of a case]. AB - A 61 years old patient with Wolff-Parkinson-White's syndrome, and hypertension was admitted to the CCU, because of the first episode of substernal chest pain. ECG was deformed by Wolff-Parkinson-White's syndrome, type B, with accessory pathway located on the right side, without evolution. Serum enzymes remained low. Echocardiography showed akinesis of the posterior wall and hypokinesis of the lateral wall (the same contraction disorders were described in previous echocardiographical examination 5 years ago), it was observed that the first portion of myocardium to contract was the base of the right ventricle. To elucidate the etiology of the contraction disorders, scintigraphy of the heart, using Thallium 201, was performed, showing normal perfusion of the myocardium. To illustrate the dependence of the contraction disorders and abnormal depolarization pattern of the heart, echocardiographical examination was repeated, confirming the previous results, then 100 mg of Ajmaline was given to the patient intravenously, and echocardiographical examination was continued. Administration of the drug caused antidromic atrioventricular re-entrant tachycardia during which the lateral wall of the heart had been contracting properly. This case shows contraction disorders of the heart caused by the abnormal depolarization pattern, resulting from the presence of accessory pathway. It also illustrates the diagnostic difficulties in patients with Wolff Parkinson-White's syndrome and suspected myocardial infarction, at the same time showing that scintigraphy of the heart might be very helpful in such patients. This case confirms the usefulness of echocardiography for localization of the accessory pathway. PMID- 8231000 TI - [New views on the pathogenesis of hypertension]. PMID- 8231001 TI - [External cardiac pacing. Today and tomorrow]. PMID- 8231002 TI - [Heparin in heart failure and unstable angina pectoris]. PMID- 8231003 TI - [Antithrombotic effects of molsidomine, isosorbide dinitrate and verapamil in a laser-induced thrombosis model]. AB - Antithrombotic effects of molsidomine, isosorbide dinitrate and verapamil obtained from the Casela Riedel Pharma GmbH, Frankfurt/Main, Germany have been studied in the laser-induced rat thrombosis model. The investigations were carried out on male Wistar rats weighing 200-300 g. Thrombus formation was induced in small mesenteric arteries--25-35 microns diameter using argon laser. An interference contrast system based on a Leitz Orthoplan microscope for the evaluation of thrombus formation was used. The number of laser injuries needed to induce a defined thrombus proved to be a useful way to quantitate the results in this thrombosis model. All agents showed dose dependent antithrombotic effect in our laser model. Molsidomine and verapamil inhibited 30 min after single i.v. injection in minimal dose 1 mg/kg and in dose 5 mg/kg 2 hours after oral administration thrombus formation in small mesenteric arteries. Isosorbide dinitrate in higher doses: 10 mg/kg 30 min after i.v. injection and 20 mg/kg after oral feeding showed significant antithrombotic effect. Antithrombotic effect of administration of minimal effective doses of all agents lasted longer than 4 hours but less than 6 hours. All drugs in effective antithrombotic doses after intravenous injection showed also inhibitory effect on rat platelet adhesion to the siliconized glass and bovine subendothelial extracellular matrix. PMID- 8231004 TI - [Hemodynamic significance of atrio-ventricular sequence in the example of an observed case]. AB - A case of a 62 year old man with chronic complete atrio-ventricular block but persistent electrical and mechanical atrial activity is presented. The patient was treated by ventricular pacing of the VVI type. During cardiac catheterization right atrium, right ventricle and pulmonary pressure were recorded at basal conditions; right ventricle contractility index and cardiac output were also estimated. An elevation of right atrial pressure simultaneously with a fall of right ventricle and pulmonary pressure were repeatedly observed. These changes were a consequence of atrio-ventricular sequence disorders resulting in un coordination of atrial and ventricular contractions. Hemodynamic parameters were restored to the physiological range only during the chance appearance of normal atrio-ventricular sequence. The presented observations support the known fact that ventricular pacing in patients with complete atrio-ventricular block may lead to serious hemodynamic disturbances. PMID- 8231005 TI - [Angioplasty of several stenoses including the left main coronary artery in a patient with acute circulatory insufficiency after myocardial infarction]. AB - A 74 year old man with diabetes and multivessel coronary heart disease involving also left main coronary artery is presented. The patient was admitted to the hospital because of acute myocardial infarction. PTCA of left main coronary artery and other four severe stenoses was attempted. The clinical status after procedure, short-term (after eight months) and long-term (after five years) was much improved. PMID- 8231006 TI - [Treatment of recent myocardial infarction during menstruation--presentation of a case]. AB - A young woman was admitted because of recent myocardial infarction (MI). Although she displayed menstruation bleeding, thrombolytic therapy with tpA was instituted. This resulted in full reperfusion and uneventful clinical course of MI. There was no prolongation of menstruation. Fibrinolytic treatment in women with MI and menstruation seems to be a safe procedure. PMID- 8231007 TI - [Left ventricular diastolic disfunction in hypertension. Effect of medications]. PMID- 8231008 TI - [Angiotensin converting enzyme inhibitors: drug of first or second choice in hypertension in advanced age]. PMID- 8231009 TI - [Transesophageal echocardiograph in the evaluation of proximal segments of coronary arteries]. AB - The study was undertaken to assess the usefulness of TEE for evaluation of morphology and flow in coronary arteries. TEE (2D, spectral and color Doppler imaging) and coronary angiography were performed in 75 patients (pts)-41 with valvular heart disease and 34 with ischemic heart disease. Proximal coronary artery stenosis was detected by coronarography in 11 pts (9--left main coronary artery, 2--right coronary artery). TEE visualization of proximal coronary arteries was possible in all pts. Echocardiographic features of artery stenosis were: the narrowing of the vessel in 2D image (9 pts), high flow velocity spectral Doppler (4 pts, mean 135 cm/s vs 55 cm/s in normal arteries) and mosaic, turbulent flow in color Doppler (10 pts). Sensitivity and specificity of TEE for coronary artery stenosis detection was respectively 81%/98% for 2D imaging and 90%/100% for color Doppler. TEE is a new, noninvasive and safe method for the evaluation of proximal coronary arteries. Detection of LMCA stenosis prior to catheterization may enhance the safety of coronary angiography. PMID- 8231010 TI - [Hypertrophic cardiomyopathy occurring in the family. Rare coexistence with oligophrenia. Clinical examinations, cytogenetic and HLA system]. AB - Familial hereditary ventricular hypertrophy (HCM) is classified as a genetically determined disease (autosomal dominant trait) characterized by generalized ventricular hypertrophy, specific heart sounds and echocardiography images, characteristic ECG changes. Sudden death occurs in some cases. Clinical data and laboratory findings in a family of twelve, in which three brothers (aged 17, 21 and 25) displayed typical features of hypertrophic cardiomyopathy, are presented. In addition to the HCM symptoms, all brothers displayed unique, characteristic phenotype: long upper and lower extremities, microcephaly and different in degree mental retardation. Echocardiography and Holter monitoring revealed types III and IV (according to Maron's classification) with complex ECG disturbances. In other members of the family the following changes were found: supra and ventricular arrhythmias appeared in the ECG of the mother (45 years old) in the forth decade of her life and ST disturbances ("silent ischaemia") in the ECG of the father (44 years old). Arrhythmias were present in the father's brother and sister, but without any clinical signs of HCM. Cytogenetic analysis was performed on the peripheral blood lymphocytes derived from the mother and all her sick sons--the karyotypes were normal. Additional cytogenetic studies detecting the presence of chromosome fra (16) were negative. Analyses of the HLA antigens were performed on 13 members of the three generations in the family. The HLA antigens of classes I A, B and C were identified and results suggest some linkage between HCM and B12 (44) antigen. To our knowledge, the present study provides the first description of a family displaying simultaneously ventricular hypertrophy and a specific phenotype with mental retardation. PMID- 8231011 TI - [Ultrasonic evaluation of chronic aortic regurgitation. Personal investigations]. AB - Decision for valve replacement in valvular heart disease is actually based on non invasive assessment without cardiac catheterization. This decision is particularly difficult in asymptomatic patients with aortic regurgitation (AR). Semiquantitative and quantitative assessment of the AR was performed in 74 pts aged 11-66, mean 34.9, 26 women, 48 men. M mode, 2D, Doppler PW, CW, color flow mapping in parasternal long, short axis, 4.5 CH and suprasternal view were obtained by Hewlett Packard Sonos 500 or 1000 with 3.5, 2.5 and 1.9 MHz transducer. LV internal diastolic diameter (LVIDD), stroke volume (SV) of the left and right ventricle, regurgitant fraction (RF), regurgitant volume (RV), regurgitant orifice (RO), LVEDP, deceleration slope, pressure half time (PHT) of regurgitant flow and proximal color flow regarding aortic valve diameter were calculated. The value of RF was 19-81%, mean 53 +/- 16% and correlated with SV (r = 0.72) Relation of the RF and LVEDP should be noticed. Positive correlation with observed up to 20-25 mmHg LVEDP. Further increasing LVEDP caused decreasing of the RF. High LVEDP confined regurgitant volume, because of diminished aorta (Ao) LV diastolic pressure gradient. Ao-LV diastolic gradient dynamics is a very important supplement of LVEDP assessment. Deceleration in this group was 147-805 cm/s2 mean 270 +/- 120. PHT 128-812 ms mean 270 +/- 161 and correlated well with LVEDP (r = 0.87 and r = 0.85). Proximal color width in parasternal long axis correlated with RF (r = 0.76) and RO (r = 0.77). LVIDD was 4.3-8.8 cm mean 6.2 cm correlated with LVEDP r = 0.60, PHT = -0.69.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231012 TI - [Sensitivity of exercise echocardiography, electrocardiographic exercise test and 24-hour ekg monitoring in diagnosing and evaluating the efficacy of coronary disease treatment]. AB - To test the sensitivity and specificity of noninvasive tests for detecting myocardial ischemia, 78 patients undergoing cardiac catheterisation for suspected coronary artery disease (CAD) were studied in prospective blinded manner with exercise echocardiography (EE), ecg stress test (ET) and 24-hour Holter monitoring for analysis of ST-segment changes (HM). All noninvasive tests were repeated after one year follow-up period. Exercise was performed in a supine position using a bicycle ergometer. Real time 2-DE examinations were performed in the left lateral position: at rest, at the peak of exercise and immediately after exercise testing. The sensitivity and specificity of EE, ET and HM for detection of CAD were: 76%, 92%; 90%, 48%; and 80%, 71% respectively. In a group of 48 pts with the narrowing of coronary artery (CA) < 50%, the follow-up EE was negative in 45. In a group of 30 pts with significant narrowing of CA, 10 were asymptomatic within one year after PTCA or CABG; in 9 of them the follow-up EE as well as ET were negative. HM less correlated with clinical course in asymptomatic subjects: in 4 patients it was negative and in 6 - positive. The reminding 20 patients with CAD treated medically, with PTCA or CABG were symptomatic after one year. The follow-up EE, ET and HM studies were positive in 17, 19 and 17 cases respectively. We conclude that sensitivity of EE in diagnosis of CAD is comparable to ET and HM, but its specificity is higher.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231013 TI - [Does effective cardioversion after surgery for valvular heart disease improve hemodynamic indicators?]. AB - In 55 patients aged 47 +/- 6 years successful electrical cardioversion of atrial fibrillation was made. 24 hours before and after cardioversion, ekg and M-mode, 2 D and Doppler echocardiography was performed. Important improvement in ejection fraction (Simpson method), (p < 0.005) and in stroke index (Simpson method, p < 0.01; Doppler method, p < 0.005) was found in 64% of pts. Pts with haemodynamic improvement (HI) were significantly younger, had smaller, left atrial area in apical two-chamber view and left ventricular diastolic diameter and longer pulmonary acceleration time. In pts with HI the duration of AF was significantly shorter and closed mitral commissurotomy less frequent (p < 0.05). Restoration of mechanical atrial function (A-wave) was found in 44% of pts. In pts with A-wave HI was more often (p < 0.001). There was no correlation between maximal peak A wave flow velocity and HI. PMID- 8231014 TI - [Echocardiographic evaluation of mitral regurgitation after myocardial infarction]. AB - Pulsed Doppler and echocardiographic analysis of the incidence of mitral regurgitation (MR) was performed. Moreover, an attempt at finding out factors conducive to the development of MR in patients after myocardial infarction (MI) was undertaken. The study included 70 patients (53 males and 17 females) aged 29 70 years, mean age: 56 years, with (first in their life) Q-wave infarction. Twenty-eight of these patients (40%) had anterior wall MI, 34 (49%) had inferior wall MI and 8 (11%) had apical MI. Pulsed Doppler echocardiographic analysis was performed 3 months after MI. In 47 patients (67%), papillary muscle dysfunction with abnormal mitral valve leaflet closure was observed. MR was found in 37 (79%) of these patients. MR diagnosed by Doppler echocardiography was found to be a frequent complication of MI (53% of patients). In echocardiographic assessment, patients with Doppler MR had a significant enlargement of mitral anulus, LV dilatation and higher percentage of asynergic basal segments of the left ventricle as compared with patients without MR. These changes were more frequently observed in patients with inferior or apical infarctions and ejection fraction < 40%. PMID- 8231015 TI - [Young men with a positive family history of hypertension have higher values in a circadian blood pressure profile]. AB - Non-invasive automatic 24-h indirect monitoring of blood pressure was performed in two groups of young males - group S - 55 men with positive family history of essential hypertension (EH) (mean age 24.6 +/- 3.6 years) and group K - 11 men with negative family history of EH (23.5 +/- 3.7 years). Circadian blood pressure variations were studied in two-hour periods throughout 24 hours. The curves of the blood pressure profiles in groups S and K had similar shape. The mean systolic blood pressure estimated for each of the two-hour periods was higher in group S than in group K in the corresponding period. Mean diastolic blood pressure estimated for each of the two-hour periods was higher in group S than in group K in almost all periods of the daily activity. These results indicate that young males with positive family history of EH have higher blood pressure than young males with negative family history of EH. This stable disregulation of blood pressure is present not only during day-time activity but also during sleep. PMID- 8231016 TI - [Transvenous ablation of recurrent ventricular tachycardia in arrhythmogenic right ventricular dysplasia]. AB - Catheter ablation was used to cure refractory ventricular tachycardias (VT) in a 20-years old lady with arrhythmogenic right ventricular dysplasia. Antiarrhythmic drugs (procainamide, amiodarone, gilurytmal, flecainide and beta-blockers) used in monotherapy or combination didn't prevent recurrence of sustained VT. During electrophysiological study 3 different morphologies of tachycardia were induced, indicating multiple sites of arrhythmia. One of them was typical for right ventricular outflow tract and similar to the VT recorded in clinical conditions. Endocardial mapping in that region showed pathological low amplitude, fragmented potentials. They preceded by 35 ms the onset of QRS complexes during VT. This area was suggested as a presumed origin of the VT and chosen for transvenous ablation. 11 direct current shock of 200-250 joules (total energy 2400 j) were delivered. No complications were seen during and after ablation. The procedure was terminated when only non sustained VT can be induced by programmed stimulation. The same results were obtained during the control study one month later. However, the patient was taking sotalol and mexiletine. During 6 months period of ambulatory observation the patient was doing well, free of arrhythmias. PMID- 8231017 TI - [Dissecting aortic aneurysm complicated by rupture of the right coronary artery and acute myocardial infarction--in a person treated with fibrinolysis]. AB - A case report of 45-year-old woman with acute inferior wall and right ventricle myocardial infarction treated with i.v. Streptokinase is presented. Few days later acute aortic dissection involving right coronary artery was detected. After six days the patient was successfully operated. She is in good condition 10 months afterwards. PMID- 8231018 TI - [Left ventricular hypertrophy is unfavorable to coronary circulation]. PMID- 8231019 TI - International dialysis aid in earthquakes and other disasters. PMID- 8231020 TI - Clinical implications of hemodialysis membrane biocompatibility. PMID- 8231021 TI - Distribution of dopamine- and cAMP-dependent phosphoprotein (DARPP-32) in the developing and mature kidney. AB - DARPP-32 is a dopamine- and cAMP-regulated inhibitor of protein phosphatase-1 (PP 1). Dopamine and DARPP-32 regulate sodium reabsorption in renal tubules by inhibiting the activity of Na+,K(+)-ATPase. We here report the pre- and postnatal distributions of DARPP-32 in the kidney as demonstrated by immunoblotting and immunohistochemistry. With immunoblotting we examined the abundance of DARPP-32 and the functionally similar but more widespread inhibitor of PP-1, inhibitor-1 (I-1). We compared their relative abundance in the renal cortex, renal medulla and neostriatum from the brain, where DARPP-32 is greatly enriched. DARPP-32 levels in the adult rat were fourfold higher in the neostriatum than in the renal medulla and 13-fold higher than in the renal cortex. I-1 levels were approximately the same in the neostriatum and in the renal medulla and 2.5-fold higher in neostriatum than in the renal cortex. Between postnatal day 10 (PN10) and 40 (PN40) DARPP-32 abundance increased 1.3-fold in the neostriatum, 1.4-fold in the renal cortex and sixfold in the medulla. The abundance of I-1 did not increase in the striatum from PN10 to PN40 but increased 1.5-fold in the renal cortex and threefold in the renal medulla. Thus, during the time of maturation of tubular transport function, the levels of both PP-1 inhibitors increased in the kidney, the largest increase being found in the renal medulla. With immunohistochemistry strong DARPP-32-like-immunoreactivity (DARPP-32-LI) was detected in the ureteral buds from gestational day 18 and up to postnatal day 8 when nephrogenesis was completed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231022 TI - Simvastatin inhibits PDGF-induced DNA synthesis in human glomerular mesangial cells. AB - Inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A-(HMGCoA) reductase ameliorate glomerular pathology and renal dysfunction in different models of glomerular disease. This effect has generally been attributed to a decrease in the circulating levels of cholesterol. Focal or diffuse mesangial cell proliferation is a common feature of glomerular pathology. There is now evidence from studies in vitro and in vivo that platelet-derived growth factor (PDGF) is an important mediator of glomerular hypercellularity. The activity of HMGCoA reductase has previously been shown to be a requirement for cell growth. In the present study, we examined the effect of simvastatin, and HMGCoA reductase inhibitor, on PDGF induced DNA synthesis and PDGF B chain gene expression in human glomerular mesangial cells. In addition, we investigated the effect of simvastatin on phospholipase C (PLC) and protein kinase C (PKC) activation stimulated by PDGF. We demonstrate that treatment of the cells with simvastatin completely inhibits PDGF-induced DNA synthesis. This inhibition is reversed by mevalonate but not by cholesterol or farnesol, two major metabolites of the mevalonate pathway. On the other hand inhibition of HMGCoA reductase does not influence PDGF-induced activation of PLC and PKC, or PDGF B chain gene expression. These data suggest that simvastatin acts at a late step in the PDGF mitogenic pathway without interfering with other early cellular responses elicited by this growth factor. These studies also raise the possibility that the ameliorative effect of HMGCoA reductase inhibitors on glomerular pathology may be mediated, at least in part, by a direct cellular effect. PMID- 8231023 TI - Ion content and cell volume in isolated collecting duct cells: effect of hypotonicity. AB - On isolated inner medullary collecting duct (IMCD) cells of the rat kidney the capability of osmoregulatory adaptation was investigated in vitro. IMCD cells were isolated by differential centrifugation at 600 mOsm (268 mM NaCl) and subsequently exposed to hypotonic buffers (300 mOsm, 118 mM NaCl). The alterations of ion content and cell volume following this change in extracellular osmolarity were studied by electron probe microanalysis and determination of intracellular water. After swelling within 40 seconds to 152 +/- 15% of control (P < 0.001; N = 9) cell volume was restored after 15 minutes. This regulatory volume decrease (RVD) was observed irrespective whether extracellular osmolarity was changed by using NaCl or mannitol as the major osmolyte. During RVD the cells lost sodium (48 +/- 11%) and chloride (14 +/- 5%), and the potassium content remained nearly unchanged. Correspondingly, sodium and chloride concentrations were progressively lowered, whereas the potassium concentration changed only transiently. RVD was diminished by 10(-4) M NPPB, 10(-3) M SITS and in the absence of HCO3-. Twenty millimoles of ouabain or 5 mM barium also inhibited RVD with little additive effect. A total of 10(-3) M amiloride and 10(-4) M bumetanide showed no effect on the hypoosmotic volume response. The experiments show that in isolated IMCD cells exposed to hypotonic conditions, rapid reversible changes in cell volume and sustained alterations in cell inorganic ion content occur, and thereby transmembrane sodium and potassium gradients are maintained. Since the loss in inorganic electrolytes does not account for RVD, the major part of volume regulation seems to occur via changes in organic osmolytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231024 TI - Alteration of mesangial response to ANP and angiotensin II by glucose. AB - To test the hypothesis that the function of glomerular mesangial cells is impaired in diabetes, we examined the responsiveness of mesangial cells cultured under high concentrations of glucose to atrial natriuretic peptide (ANP1) and angiotensin II (Ang II). The ANP-induced accumulation of cGMP was enhanced in mesangial cells cultured under high glucose conditions, possibly due to the activation of particulate guanylate cyclase. Ang II action in mesangial cells was evaluated by measuring the ability of Ang II to inhibit ANP-induced cGMP accumulation through both activating phosphodiesterase (initial phase) and inhibiting guanylate cyclase (maintenance phase). The inhibition of both ANP induced cellular cGMP accumulation and particulate guanylate cyclase activity by Ang II was significantly reduced in mesangial cells cultured under high concentrations of glucose. Moreover, in the cells exposed to high concentrations of glucose, both basal and Ang II-stimulated levels of inositol 1,4,5 trisphosphate (IP3) were significantly reduced. These results indicate that, in high glucose conditions, the actions of ANP and Ang II are modulated differently, resulting in the impairment of contractile responsiveness of mesangial cells. PMID- 8231025 TI - Aldosterone modulates PNA binding cell isoforms within renal collecting duct epithelium. AB - To investigate the differentiation of the ampullary collecting duct cells into adult principal and intercalated cells, the embryonic cortex of newborn New Zealand rabbit kidney was isolated and brought in culture. With this culture technique the ampullary cells formed a polarized collecting duct epithelium which was kept under permanent exchange of medium and in the presence of aldosterone, arginine vasopressin and/or insulin. After 14 days of perfusion culture the epithelia showed light and dark cells resembling the principal and intercalated cells of the adult collecting duct. The differentiation from embryonic into adult collecting duct cells was controlled by applying the monoclonal antibody CD 7. Independent of the hormonal treatment all of the epithelial cells matured in culture and expressed the CD 7 antigen. This corresponded with the situation found within the adult kidney, where the CD 7 antigen was localized in all principal and intercalated (IC) cells, whereas the embryonic ampullary epithelium in the neonatal kidney remained negative. A differentiation feature of the beta type intercalated cell was investigated by labeling the cultured epithelia with peanut agglutinin (PNA). In contrast to the CD 7 antigen the development of PNA binding was highly dependent of time and individual hormone administration. While in control epithelia only 8% of PNA positive cells were found, aldosterone induced epithelia revealed 72% PNA labeled cells. The combination of aldosterone and insulin increased the number of PNA-positive cells to 90%. By scanning electron microscopy it could further be shown that several isoforms of cells were reactive with PNA. Thus, in culture the PNA label is not restricted to the typical beta-type IC cells. PMID- 8231026 TI - Bradykinin causes selective efferent arteriolar dilation during angiotensin I converting enzyme inhibition. AB - We studied the effects of interruption of the renin-angiotensin system (RAS) in rats that were volume depleted by water deprivation for 48 hours (AWD) with/without furosemide (AWD + F), a condition known to activate RAS. Following baseline micropuncture, AWD rats (N = 6) were treated with a specific angiotensin II type 1 receptor antagonist (AIIRA; 4 mg/kg body wt bolus i.v. and then continuous infusion) and glomerular hemodynamics compared to those obtained during angiotensin I converting enzyme inhibitor treatment (ACEI; 24 mg/kg bolus i.v. and then continuous infusion). Systemic blood pressure decreased equally following AIIRA and ACEI. Single nephron glomerular filtration rate (SNGFR) increased from baseline following AIIRA (24 nl/min vs. 30, P < 0.025). While a decrease in efferent arteriolar resistance (RE) reduced glomerular capillary pressure (PGC; 67 mm Hg vs. 60, P < 0.05), this change in RE together with decrease in afferent arteriolar resistance (RA), enhanced glomerular plasma flow rate (QA; 80 nl/min vs. 111). Antagonizing angiotensin II receptor increased QA which, together with the tendency to increase glomerular capillary ultrafiltration coefficient, Kf, served to improve glomerular filtration. By contrast, although inhibition of the angiotensin I converting enzyme caused greater vasodilatation, no increase in SNGFR occurred. The lack of response in filtration after ACEI was due to a further fall in PGC to 52 mm Hg (P < 0.01 vs. AIIRA), reflecting profound reduction in RE. Since ACEI but not AIIRA potentiates bradykinin activity we examined effects of a specific bradykinin antagonist (Hoe).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231027 TI - Effect of 22-oxa-calcitriol on calcium metabolism in rats with severe secondary hyperparathyroidism. AB - The purpose of the present study was to examine the effect of a two day and a five day administration of 22-oxa-calcitriol (OCT) on calcium metabolism in rats with advanced chronic renal failure and severe secondary hyperparathyroidism. A first series of 27 uremic rats received either placebo, OCT or calcitriol (0.3 microgram i.p./rat) 48 and 24 hours before sacrifice. A second series of 18 uremic rats received either placebo, OCT (0.3 microgram i.p./rat) or calcitriol (0.05 microgram i.p./rat) for five days. We found that after 48 hours (series 1) both calcitriol and OCT increased blood ionized calcium (Ca2+) as compared to vehicle (1.23 +/- 0.04 and 1.10 +/- 0.02 mM, P < 0.01 and P < 0.05, respectively vs. control, 1.02 +/- 0.03 mM). Duodenal Ca transport (S/M) using the everted gut sac technique was not stimulated by OCT, even though it increased from 2.8 +/- 0.4 to 7.0 +/- 0.6 (P < 0.01) with calcitriol. In contrast, duodenal calbindin D9k mRNA expression and protein content increased to a similar extent with OCT and calcitriol. Calcitriol was more potent in reducing plasma iPTH1-34 levels than OCT: 344 +/- 75 pg/ml (calcitriol) versus 632 +/- 46 pg/ml (OCT) compared with 897 +/- 74 pg/ml (control), P < 0.01. In the second series of rats, the injection of OCT (0.3 microgram i.p./rat) over five days was less effective than the lower dose of calcitriol (0.05 microgram i.p./rat) in reducing circulating iPTH: 110 +/- 26 (calcitriol) and 281 +/- 64 (OCT) versus 624 +/- 135 pg/ml (control), P < 0.01.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231028 TI - Glomerular fibrinolytic activity in anti-GBM glomerulonephritis in rabbits. AB - Fibrin is an important mediator of injury in severe proliferative forms of glomerulonephritis (GN). Normal glomeruli express fibrinolytic activity, which may protect against the injurious effects of fibrin deposition. Changes in glomerular fibrinolytic activity (GFA) may play an important role in modulating fibrin accumulation in GN. To study the changes in GFA associated with fibrin deposition in GN, autologous phase anti-glomerular basement antibody initiated GN (anti-GBM GN) was studied in rabbits. Net GFA was significantly reduced in association with glomerular fibrin deposition (1.3 +/- 0.8 ng fibrin lysed/10(3) glomeruli/2 hr, normal 57.1 +/- 25.4 ng fibrin lysed/10(3) glomeruli/2 hr, P < 0.02). Reduced GFA in fibrin associated GN was associated with decreased expression of tissue type plasminogen activator (tPA) and increased expression of plasminogen activator inhibitor type-1 (PAI-1) and glomerular macrophage infiltration. In a fibrin independent model of anti-GBM induced GN (heterologous phase), with equivalent injury (proteinuria), net GFA was increased (174 +/- 64 ng fibrin lysed/10(3) glomeruli/2 hr). This was associated with increased tPA and uPA, and decreased PAI-1 in the absence of significant macrophage infiltration. These studies demonstrate that fibrin deposition in GN is associated with a net reduction of GFA, attributable to reduced expression of plasminogen activators and augmentation of PAI-1. Reduction of GFA may potentiate glomerular fibrin deposition and consequent glomerular injury. The association between glomerular macrophage influx and reduction in GFA suggests that this change may be directed by macrophages. PMID- 8231029 TI - A protein with anion exchange properties found in the kidney proximal tubule. AB - One important mechanism for reabsorption of chloride in the kidney proximal tubule involves anion exchange of chloride for a base. Anion exchange transport systems in general demonstrate sensitivity to inhibition by disulfonic stilbenes, probenecid, furosemide, and the arginyl amino group modifier phenylglyoxal. Using disulfonic stilbene affinity chromatography, we have identified and partially purified a protein with anion exchanger properties in luminal membrane vesicles isolated from rabbit kidney cortex. This protein has a molecular weight of 162 kD. The binding of the 162 kD protein to the stilbene affinity matrix is inhibited by disulfonic stilbenes, probenecid, furosemide, and phenylglyoxal. Reconstitution of the proteins eluted from the affinity matrix into liposomes demonstrates anion exchange activity as assayed by radiolabeled chloride influx. Deletion of the 162 kD protein from the eluted mixture by probenecid diminishes the anion exchanger activity in the reconstituted liposomes. Further purification of the disulfonic stilbene column eluant by Econo-Pac Q ion exchange chromatography resulted in significant enrichment in 162 kD protein abundance and also anion exchange activity in reconstituted liposomes. The results of the above experiments strongly suggest that the 162 kD protein is an anion exchanger. Insight into the functional and molecular characteristics of this protein should provide important information about the mechanism(s) of chloride reabsorption in the kidney proximal tubule. PMID- 8231030 TI - Increased Na/H antiport activity and abundance in uremic red blood cells. AB - Alterations in red blood cell sodium (Na) transport have been described in chronic renal failure. This study examines the possible impact of uremia on two ouabain-insensitive pathways, the Na/H antiporter and the Cl-/NaCO3- anion exchanger. The Vmax of Na/H antiporter measured as Na influx driven by outward H gradient in acid loaded red blood cells was significantly higher in uremic red blood cells versus controls (60.5 +/- 16.5 vs. 24.5 +/- 5.4 mmol/liter cells/hr, P < 0.025). This increase in activity was associated with an increased abundance of the Na/H antiporter as determined by immunologic analysis using an affinity purified polyclonal antibody to the human NHE-1 isoform of the antiporter. By contrast, the activity of the anion exchanger measured as the DIDS-sensitive lithium (Li) influx was similar in uremic versus control red blood cells (2.10 +/ 0.18 vs. 2.14 +/- 0.20 mmol/liter cells/hr). These experiments, when considered in conjunction with prior studies showing normal Na/Li countertransport in uremia indicate that there is a selective increase in the number of functional Na/H antiporters in uremic red blood cells and that Na/Li countertransport measurements may not be a valid marker for Na/H antiporter activity in red blood cells in patients requiring dialysis for end-stage renal failure. PMID- 8231031 TI - Dissociation between the course of the hemodynamic and antiproteinuric effects of angiotensin I converting enzyme inhibition. AB - Angiotensin I converting enzyme inhibition (ACEi) has been shown to lower urinary protein excretion in human renal disease. The mechanism of this antiproteinuric effect is hypothesized to be mediated by changes in renal hemodynamics. However, clinical studies suggest that the effect on renal hemodynamics is fully established immediately after the start of treatment, whereas others show the antiproteinuric effect to reach maximum only after several weeks. To clarify this issue we studied the course of renal hemodynamics, blood pressure and proteinuria during 28 days of ACEi (enalapril 10 mg oid) in nine patients with proteinuria due to non-diabetic renal disease. The effect of ACEi on blood pressure and renal hemodynamics was already maximal within few hours after start of treatment, and remained stable thereafter: MAP was lowered with 8.6 +/- 1.9%, 10.6 +/- 2.1%, 12.8 +/- 2.3% and 12.9 +/- 2.5%, while FF fell 23.0 +/- 2.0%, 17.0 +/- 2.6%, 16.8 +/- 2.8% and 15.9 +/- 4.0% on days 1, 7, 14 and 28 of ACEi, respectively. However, the antiproteinuric effect only gradually reached its maximum on day 28. Urinary protein excretion decreased with 10.9 +/- 6.1%, 32.7 +/- 6.2%, 46.3 +/- 2.5% and 54.0 +/- 2.5% on days 1, 7, 14 and 28 of ACEi, respectively. After drug withdrawal all parameters returned towards baseline. We conclude that a dissociation occurs in the course of the ACEi induced effects on hemodynamics and urinary protein excretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231032 TI - Interleukin-6 and TNF alpha production in human renal cell carcinoma. AB - Several clinical and biological features suggest that cytokines implicated in the inflammatory response are produced by renal cell carcinoma (RCC). To define if alterations of tumor necrosis factor alpha (TNF alpha), interleukin 1 alpha (IL-1 alpha), IL-1 beta and IL-6 gene expression are present in this malignancy, samples from 19 tumors as well as samples from seven paired normal renal tissue were examined using Northern blot and immunohistochemical analysis. In addition, the expression of these cytokines was evaluated in seven RCC-derived cell cultures using Northern blot or RT-PCR. TNF alpha and IL-6 proteins were measured in culture supernatants using specific bio- and immunoassays. Consistent levels of IL-6 mRNA were detected in 17 of the 19 tested tumors whereas TNF alpha specific transcripts were present in seven of eight available RNA samples. TNF alpha and IL-6 mRNA were also detected in five of the seven paired normal kidneys. By immunolabeling, IL-6 antigen was not detected in RCC cells in any of the 19 studied samples. In contrast, using anti-TNF alpha antibody a strong labeling of stromal endothelia and macrophage cells was detected in all the 19 cases, and evident TNF alpha staining of the carcinoma cells themselves was observed in eight cases. Spontaneous IL-6 mRNA expression was detected in five RCC cell cultures and TNF alpha mRNA in four. The cultured cells exhibited positive TNF alpha immunolabeling in six of seven cases but were always IL-6 negative. Bioactive IL-6 was detected in all culture supernatants while bioactive TNF alpha was not detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231033 TI - Collagen studies in newborn rat kidneys with incomplete ureteric obstruction. AB - Collagen studies in newborn rats with incomplete ureteric obstruction were performed to describe and quantify changes in collagen deposition resulting from urinary tract obstruction at an early developmental age. Incomplete ureteric obstruction was created in three-day-old rats by placing the left ureter in a tunnel formed by the psoas muscle, and sham-operated controls underwent a laparotomy. The rats were sacrificed at 10, 17, 24 or 31 days. Collagen types I, III, IV, and V were localized by indirect immunofluorescence microscopy, the total collagen content of the kidney was quantitated using hydroxyproline analysis, and collagen types I and III were quantitated using cyanogen bromide (CNBr) peptide analysis. Increased immunofluorescent staining for all of the collagens was found in the diffusely widened medullary interstitium of the obstructed kidney, and more focally in the cortical interstitium. Collagen types I, III and V, but not collagen type IV, were also found in bands in the interstitium at the junction of the cortex with the medulla. Increased staining for collagen type IV was found in thickened and tortuous tubular basement membranes (TBM) of the obstructed kidneys. The total collagen content of the obstructed kidney was significantly increased compared to the amounts in both the contralateral kidneys and in the kidneys from sham-operated controls at 24 and 31 days of age (P < 0.01 in each case, Wilcoxon matched pairs rank sum test and Mann Whitney U-test, respectively). The amount of collagen in the kidneys correlated with the degree of hydronephrosis (Spearman correlation test, r = 0.78, P < 0.02). CNBr peptide analysis demonstrated that over 50% of the collagen in the normal neonatal rat kidney was collagen type I and approximately 25% was collagen type III. In the obstructed kidneys most of the collagen was also collagen type I and collagen type III, although the proportion of total collagen comprised by these collagen types was decreased compared with the controls. The amount of collagen type III in the contralateral kidneys was reduced compared to that in the controls. Thus, the neonatal renal response to obstruction resulted in increased amounts of a range of collagens in the interstitium and TBM, and the extent of this response was partially related to the degree of hydronephrosis. PMID- 8231034 TI - Effects of cooler temperature dialysate on hemodynamic stability in "problem" dialysis patients. AB - Symptomatic hypotension is a common and disabling complication of hemodialysis treatments. The incidence of symptomatic hypotensive episodes is particularly high in patients who have normal or low blood pressure at the initiation of dialysis and in patients who have large interdialytic weight gains. The aim of this study was to determine whether cooling the dialysate temperature from 37 degrees C to 35 degrees C improved tolerance to dialysis in a group of 12 of these "problem" patients. A double-blinded protocol was performed in six hypotension-prone and six large weight gainers who were subjected to two identical hemodialyses except for the dialysate temperature of 37 degrees C or 35 degrees C. Changes in biochemical parameters and weight were comparable during the two maneuvers. Recumbent blood pressure declined significantly (P < 0.01) during 37 degrees C dialysis but not 35 degrees C dialysis; blood pressure was significantly lower at 1, 2, and 3 hours of 37 degrees C dialysis compared to 35 degrees C dialysis (P < 0.05). Further, both supine and upright blood pressure was significantly lower following 37 degrees C dialysis (P < 0.02). This lower blood pressure was present in both subgroups of patients. All 18 episodes of symptomatic hypotension noted during the study occurred during 37 degrees C dialysis. A significantly greater increase in peripheral vascular resistance (calf blood flow was measured directly by venous occlusion plethysmography) occurred upon exposure to the 35 degrees C dialysate in both subgroups of patients (P < 0.01); supine and upright post-dialysis plasma norepinephrine values were also significantly greater (P < 0.001) after 35 degrees dialysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231035 TI - Association of cyclosporin A with acquired cystic kidney disease of the native kidneys in renal transplant recipients. AB - Acquired cystic kidney disease (ACKD) is a common complication in patients treated with long-term dialysis. Previous studies performed prior to the cyclosporin A (CsA) era indicate that successful renal transplantation causes regression of ACKD. Little has been published, however, on the occurrence of ACKD in CsA-treated transplant recipients. We conducted a prospective sonographic study in 33 renal transplant recipients and 32 dialysis patients to evaluate the effect of CsA on ACKD in transplant recipients. Transplant recipients had a lower prevalence (39% vs. 56% in dialysis patients) and severity (smaller kidneys and lower cyst grades) of ACKD when compared with dialysis patients. Renal cell carcinoma was found in two dialysis patients. Using multiple regression analysis, we found that the use of CsA was significantly correlated with the presence of ACKD in transplant recipients (57% in CsA-treated vs. 8% in non-CsA-treated patients). We conclude that renal transplantation reduces the prevalence and severity of ACKD in the native kidneys; however, among transplant patients, CsA administration is associated with a greater prevalence of ACKD. PMID- 8231036 TI - Screening diabetic transplant candidates for coronary artery disease: identification of a low risk subgroup. AB - Coronary artery disease is the major cause of death in diabetic renal transplant recipients. Because one-third of diabetic transplant candidates have clinically silent coronary artery disease, many transplant centers recommend coronary angiography prior to transplantation. However, angiography is expensive and may precipitate acute renal failure. Therefore, we developed a noninvasive screening algorithm to identify patients at low risk for coronary artery disease (CAD), defined as one or more coronary stenoses > or = 50% diameter. We performed coronary angiography in 141 consecutive asymptomatic Caucasian type I diabetic renal transplant candidates. Fourteen of 16 patients age 45 or older had CAD. One hundred and twenty-five patients under age 45 were randomly divided into two groups. Ninety patients were used to identify clinical factors significantly associated with CAD which included smoking for five or more pack years, nonspecific ST-T wave changes on electrocardiogram, and diabetes duration 25 years or longer. The screening algorithm, "CAD is predicted in diabetic transplant candidates under age 45 with any of the above risk factors," was then tested in the remaining 35 patients and in 35 additional patients. In these 70 patients, the algorithm had a sensitivity of 97% and a negative predictive accuracy of 96%. We conclude that coronary angiography should be recommended to Caucasian type I diabetic renal transplant candidates age 45 or older because of the high probability of disease. In patients younger than 45 without a smoking history, ST-T wave changes on EKG, or diabetes longer than 25 years, the likelihood of CAD is low and angiography can be avoided. PMID- 8231037 TI - Urea kinetics during hemodialysis measured by microdialysis--a novel technique. AB - A microdialysis technique has been developed for estimation of concentrations of low molecular size compounds in the interstitial fluid in vivo. With this technique urea kinetics in the interstitial fluid and plasma were studied in ten patients during and after hemodialysis. There was a close correspondence between urea measurements in plasma and interstitium during hemodialysis. Urea rebound occurred in plasma during two hours after dialysis (15.8 +/- 6.5% in the first hour and 11.8 +/- 5.9% in the second hour). The urea rebound in the interstitium was delayed about 60 minutes after that of plasma (2.8 +/- 8% and 14.1 +/- 7.8% in the first and second hours, respectively) and continued for up to four hours after dialysis. The relationship between plasma urea rebound and the efficiency of hemodialysis and ultrafiltration volume was studied in 17 patients. Results showed a close relation between the fractional urea removal during dialysis and the plasma urea rebound. The contribution of de novo urea genesis to the rebound was estimated from the interdialytic increase in plasma urea concentrations, and was 17 to 24% of the plasma urea rebound during two hours postdialysis. The initial plasma urea rebound could in part result from urea influx to plasma from the enterohepatic recirculation of urea nitrogen. Plasma urea rebound should be taken into account for determination of the amount of dialysis delivered during hemodialysis. PMID- 8231038 TI - Derangements in acetylcholine metabolism in brain synaptosomes in chronic renal failure. AB - Chronic renal failure causes abnormalities in the central nervous system function and in norepinephrine metabolism of brain synaptosomes. The present study examined the effect of renal failure on the metabolism of another neurotransmitter, acetylcholine, which is involved in the modulation of behavioral and motor function. We measured acetylcholine content and release, choline content, uptake and release and activity of choline kinase in synaptosomes from rats with renal failure with various duration, renal failure parathyroid-ectomized rats maintained normocalcemic, renal failure and normal rats treated with verapamil. Acetylcholine content increased while choline content decreased proportionally and significantly (P < 0.01) with the duration of renal failure; choline kinase activity was reduced (P < 0.01). These derangements were prevented by parathyroidectomy of renal failure rats or by their treatment with verapamil. Choline uptake and release were elevated in renal failure and these abnormalities were not corrected by parathyroidectomy or verapamil therapy. Acetylcholine release was elevated in renal failure and parathyroidectomy prevented this derangement. Verapamil reduced acetylcholine release in both normal and renal failure rats. The data show that: (a) renal failure causes significant derangements in acetylcholine metabolism leading to its accumulation in and an increase in its release from brain synaptosomes; (b) this is mainly due to reduced activity of choline kinase, most likely, mediated by the state of secondary hyperparathyroidism of renal failure; (c) blocking the parathyroid hormone-induced calcium influx into synaptosomes by verapamil prevented the abnormalities in acetylcholine metabolism; and (d) the derangement in choline uptake and release in CRF is not related to excess parathyroid hormone since parathyroidectomy or verapamil treatment did not correct them. PMID- 8231039 TI - The increased risk of coronary heart disease associated with nephrotic syndrome. AB - Patients with nephrotic syndrome (NS) are believed to be at increased risk of atherosclerosis and coronary heart disease (CHD), although existing evidence for this association has not been persuasive. The risk of CHD among 142 persons with NS documented by protein-uria > or = 3.5 g daily was compared with that among 142 matched controls randomly selected from the membership of a large Northern California health plan. Controls were matched for sex, year of birth, and presence in the health plan when the referent case was diagnosed. No diabetics were included in this study. Mean follow-up for nonfatal CHD events was 5.6 years for NS subjects and 11.2 years for controls. Among the NS subjects myocardial infarction (MI) developed in 11, and there were 58 deaths, seven because of CHD. Among the controls, there were four MIs and 10 deaths, three because of CHD. In matched-pair analysis, there were 11 MIs among NS subjects and none among controls [P = 0.001, lower bound of 95% confidence interval for relative risk (CI), 2.8]. In an unmatched analysis adjusted for hypertension and smoking at diagnosis of NS, the relative risk of MI was 5.5 (95% CI 1.6 to 18.3) and the relative risk of coronary death was 2.8 (95% CI 0.7 to 11.3). Omitting data of NS subjects with minimal change disease and systemic lupus erythematosus yielded similar results. These data suggest that persons with NS are at increased risk of CHD. PMID- 8231040 TI - Renal complications of nonsteroidal anti-inflammatory drugs. PMID- 8231041 TI - [Prospective study of cataract after 5 years intraocular lenses with and without laser ridge]. AB - BACKGROUND: A prospective randomized study was carried out to investigate whether intraocular lenses with and without laser ridge have different effects on the development of secondary cataract. PATIENTS AND METHODS: An extracapsular cataract extraction with implantation of a posterior chamber lens was carried out in the period from November 1986 to March 1987 in 100 patients (100 eyes). The first group of 50 patients (50 eyes) received a convex-plane (plano-posterior) intraocular lens (IOL) with a continuous 360 degrees-laser ridge. The second group of 50 patients (50 eyes) received a convex-plane (plano-posterior) IOL without laser ridge. The follow-up investigations were carried out after 10 days, 2 months, 6 months and 1, 2, 3, 4 and 5 years. Parameters such as the state of intraocular irritation, the intraocular pressure and astigmatism were also registered during the first six months after the operation. Attention was paid to the opacity of the posterior capsule and the deterioration in visual acuity associated with this in the subsequent investigations 1, 2, 3, 4 and 5 years after the operation. All cases with a deterioration of vision by two lines caused by an opacity of the posterior capsule were evaluated as secondary cataract. RESULTS: On the basis of this classification, the rate of secondary cataract in the IOL group without laser ridge was 18.5%, as compared to 19.2% in the IOL group with laser ridge. CONCLUSIONS: After 5 years there is no statistical significant difference between the two groups (Fishers-Exact-Test, p = 0.999). PMID- 8231042 TI - [Tarsal flap technique. A technically simple procedure for correction of lower lid flaccidity, senile ectropion and ptosis of the lateral canthus]. AB - Lower lid flaccidity, senile extropion and canthal malposition are the principal indications for the tarsal flap technique. This is a technically simple procedure avoiding sutures on the border of the lashes and avoiding the disadvantages of the technique of Kuhnt - Szymanowski, which cause a shortening of palpebral length due to tarsal resection. PMID- 8231043 TI - [Age dependence of pupillary near reflex]. AB - BACKGROUND: This study was performed to gain age correlated normal values for the pupillary near reflex. METHODS: Accommodation and pupillary near reaction were measured by means of simultaneous infrared video retinoscopy and pupillography in 64 healthy volunteers aged between 5 and 55 years. Measurements were done at a reduced, near mesopic, light condition with accommodation to 10, 14, 20 and 33 cm. RESULTS: The pupillary near response varied highly with age: persons younger than 20 years of age showed a significantly smaller pupillary near response as compared to those older than 20 years. In most of the children younger than 10 years the pupil near response was very small (less than 10% constriction) at accommodation distances longer than 10 cm. There was a significant difference between the age groups younger and older than 20 but no statistically significant differences within these age groups. CONCLUSION: A change of the pupillary near reaction takes place around the age of 20. We conclude that this change does not only reflect the aging of the cristalline lens but is due to an age related change of the supranuclear control. PMID- 8231044 TI - [Melanoma of the iris with recurrent hyphema--course over 32 years]. AB - BACKGROUND: The differential diagnosis of melanocytic iris tumours is very difficult. Signs of malignancy such as pupil distortion, ectropium uveae, secondary cataract and secondary glaucoma can also occur with benign iris nevi. However, spontaneous hyphemata occur in 6.9% to 13% of melanomas, while anterior chamber hemorrhages are extremely rare with nevi. PATIENT AND METHOD: A 69 year old man with a pigmented iris tumor of the right eye strictly refused surgical treatment. Therefore the tumor was followed clinically over 32 years. Over this period the tumor showed a very slow increase in size. Twenty-seven years after the initial presentation for the first time a spontaneous anterior chamber hemorrhage occurred. In the following period, the patient experienced recurrent hyphemata at last approximately every two months. An excisional biopsy consisting of a sector-iridectomy and localized excision of Descemet's membrane was performed. Histologically, the tumor was classified as a melanoma of the iris. CONCLUSION: A careful documentation cannot be overemphasized in the follow up of patients with pigmented iris lesions. In case of spontaneous hyphema histological evaluation is required even in the absence of evident tumor growth. PMID- 8231045 TI - [Prognosis of intralenticular lead foreign bodies]. AB - BACKGROUND: The prognosis of intraocular foreign bodies consisting of lead remains controversial. While experiments have shown that infected lead becomes sterile when fired from rifles, both the total absence as well as presence of severe inflammation have been observed following perforating injuries with lead. Furthermore, alterations of the ERG have been described in experimental settings. This case report is presented in order to clarify the long term prognosis of intralenticular lead particles. PATIENT AND METHOD: The 69-year-old patient suffered from a bilateral perforating injury caused by a bullet during World War II (1942). The right lens was extracted due to a secondary cataract several weeks following the injury. The left lens remained clear despite a paracentrally located metallic foreign body. The patient underwent extracapsular cataract extraction in 1992 because of an opacity of the posterior capsule which was well separated from the foreign body. Preoperatively the ERG was normal. An energy dispersive X-ray microanalysis in a scanning electron microscope was performed in order to determine the chemical composition of the foreign body, and X-ray diffraction with a Gandolphi camera was used for its structural characterisation. The foreign body consists of lead. However, its surface is probably altered to lead oxide, hydroxide or carbonate, which hindered further reaction of the lead. Calcium phosphate was deposited on the walls of the foreign body. CONCLUSION: Our report shows that lead in the lens remains inert for several decades and does not cause alterations in the ERG. Therefore, intralenticular lead outside of the optical axis may be left in situ unless a secondary cataract develops. PMID- 8231046 TI - [Primary manifestation of Burkitt's lymphoma of the non-African type in the orbits]. AB - BACKGROUND: Primary orbital involvement in non-African Burkitt's lymphoma does not occur frequently but when it occurs children are mostly affected. PATIENT AND METHOD: A 83-year-old woman with a primary manifestation of non-African Burkitt's lymphoma in the orbit was presented in our out-patient department. RESULTS: After radiation therapy no recurrence after 6 months. CONCLUSION: To our knowledge, this is the first reported adult patient with definite primary involvement of the orbit in non-African Burkitt's lymphoma. PMID- 8231047 TI - [Stromal iris cyst]. AB - A 50-year-old female developed a cyst of the iris stroma in her right eye. Because of slight enlargement within 10 weeks sector iridectomy was performed. Histology showed that the cyst was lined by a cellular layer resembling conjunctival or corneal epithelium. The benign clinical course (normal function, normal intraocular pressure and no recurrence after 4 years) and the intrastromal rather than epistromal localization hold evidence for a spontaneous origin. Spontaneous cysts of the iris stroma especially in adults are rarely described in the literature. The question still to be answered is whether these lesions are really spontaneous in origin or whether they represent only an intraocular epithelial invasion due to an occult perforation of the eye. PMID- 8231048 TI - [Comparison of phacoemulsification and planned extracapsular cataract extraction]. AB - 40% of cataract surgeons in German speaking countries use phacoemulsification as the procedure of choice for cataract extraction while the other 60% still use the planned extracapsular extraction. The more procedures a surgeon performs the stronger is the tendency toward performing phacoemulsification. The opening technique of the anterior capsule is chosen accordingly: for phacoemulsification capsulorhexis is preferred, while for the planned extracapsular extraction can opener - or letter box - technique is usually done. The length of the sclero corneal incision is 3.2 mm in phacoemulsification, 6.2 mm for a PMMA posterior chamber lens implantation and 9-13 mm for the planned extracapsular extraction. We prospectively compared both procedures with each group including 100 consecutive patients. The mean irrigation volume was 96.6 +/- 48.8 ml in the phacoemulsification group as opposed to 48.1 +/- 26 ml in the planned extracapsular extraction group. The mean duration of irrigation was 130 +/- 67 sec in the first versus 129 +/- 76 sec in the second group; mean endothelial cell loss was 7.9% versus 7.1%. No correlation was found between endothelial cell loss and duration of irrigation. Mean postoperative astigmatism measured 1.0 +/- 0.49 in the first group as compared to 3.3 +/- 1.7 dpt in the second group. Except for the astigmatism, our study could not detect significant differences regarding the outcome between both procedures. There are advantages to the phacoemulsification procedure, however, such as a smaller incision (e.g. tunnel technique), a closed irrigation-aspiration system, controlled intraocular pressure and the more frequent use of capsulorrhexis. Preferred indications and possible advantages regarding intra- and postoperative complications are discussed. PMID- 8231049 TI - [Simple rapid contrast sensitivity test in diagnosis of glaucoma]. AB - BACKGROUND: Psychophysical tests employing high-frequency stimuli are becoming of increasing value in glaucoma diagnosis. In the present study a simple, quick, and sensitive flicker test is described which can be used as an screening examination for glaucoma. METHOD: A flicker test is described which uses a white sinusoidally flickering light presented at 37.1 Hz in a ganzfeld bowl under photopic conditions (ca 10 cd/m2. Fication is not necessary. The modulation (temporal contrast) of the flicker light is varied in small steps until flicker threshold is reached. The reciprocal of the threshold value represents the temporal contrast sensitivity. The test takes only about 5 min per eye. RESULTS: 69 eyes of 69 normals, 60 eyes of 60 ocular hypertensive (OHT), and 50 eyes of 50 patients with primary open angle glaucoma were statistically evaluated. The validation of the sensitivity in both eyes was 75.6% and the specificity was 85.9%. 41.7% of the OHT patients were classified as pathologic. CONCLUSIONS: Thus, the ganzfeld flicker test is a simple and quick (5 min) additional screening test in glaucoma. PMID- 8231050 TI - [Alterations in bowel elimination: constipation]. PMID- 8231051 TI - [Altered patterns of urinary elimination]. PMID- 8231052 TI - [Stoma care]. PMID- 8231053 TI - Endotracheal suctioning (ETS) in premature infants. PMID- 8231054 TI - [A study of enhancing the image of nursing: action plan, implementation and evaluation]. AB - The purpose of this study was to develop a strategy for the promotion of the image of nursing. The study questions were: Do nurses have a proper self image? What image of nursing do the public have? It is thought that the prejudices that the public have about nursing personnel have to be eliminated in order to provide for better health care. Even though the public have misconceptions of prejudices, nurses have not paid much attention to them, nor sought ways to change them. This study was designated to make out a model project to improve the image of nursing held by the public. This study was a strategy building descriptive study. This study was oriented to a model project to improve the image of nursing. The subjects for the study were 650 nurses who were staff nurses. The study procedures were as follow: First step: a special action committee for nursing image making was established of nine members who were divided into five subgroups. 2nd step: a 1st workshop was held to improve self concept of nurses and to recognize them the necessity of nursing image development, a 2nd workshop was held to develop a conceptual framework for the action plan and for budget planning. 3rd step: a master plan for a nursing image was developed and evaluated through discussion and presentation. 4th step: lecture and role playing were used to further the development of a caring attitude in the nurse.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231055 TI - [A comparative study of the opinion on the image of nursing among the doctors, nurses, other hospital personnel and the general public]. AB - The hospital is not the place for functional isolationalism, that is, for each department to do their own activities independently. Rather it is an organization in which each part is joined together and concentrating on the same objectives. The hospital must maintain a one-system-function, so that all the resources of the hospital may be used efficiently. Although we do not yet completely understand how the nursing image as perceived by the medical team and the other members of the interdisciplinary team affects nursing practice. It is important to identify how the client recognizes the nursing image as this will influence the growth of the nursing profession directly and indirectly. So we must reevaluate the nursing image as presented to the client and find the ways to present them with a better image of nursing. The study procedures were as follow: First, the image of nurses was analyzed. The subjects for this study were 270 nurses, 172 doctors, 237 hospital employees, and 240 patients and their families from 5 local branches of the Yonsei Medical Center. The four negative responses shown in this study are as follows: 1) Nurses don't listen to patients. 2) Nurses are self-centered. 3) Nurses and careless. 4) Nurses are mechanical. Second, the concrete causes of the four negative responses were analyzed. Third, the contribution of the nurses for clinical practice, research and education was analyzed. Data were collected by questionnaires all seps of the research. Data were analyzed by use of SPSS/PC+. In conclusion, the image of nurses differed according to which group, the respondents belonged. Generally, the public have a positive image of nurses, but hospital employees have a negative images of nurses. Further the nurses gave themselves a low grade. They showed negative as to the value of their job, professionality and their contribution to clinical practice when it was compared to that of the doctors. PMID- 8231056 TI - [Nursing and law (II). Laws related to health and medicine for the nurse]. PMID- 8231057 TI - [Care of the elderly sick in the UK (welfare and policy for the aged); the challenge to nursing education]. PMID- 8231058 TI - [Development of a model baccalaureate nursing program]. PMID- 8231059 TI - [Nursing and law (II). An amendment and the changes of medical laws related to nursing]. PMID- 8231060 TI - Early gastric cancer associated with synchronous multiple liver metastasis--two rare cases. AB - From 1976 to 1991, 1,640 cases of gastric cancer were resected in our department. Of these, 659 (40.2%) cases were early gastric cancer (EGC). The prognosis after resection of an EGC is generally good, with a 5-year survival rate of 95%. The incidence of an EGC with synchronous liver metastasis is very rare. To our knowledge, only 15 cases of an EGC with synchronous liver metastasis have been reported in the Japanese literature. From 10 cases adequately described the EGC with synchronous liver metastasis derived only from sm-cancers (cancers confined to the submucosa) of medullary type, histologically, more commonly occurring in males than EGC generally, (of elevated type) and with positive lymph node metastasis. Here we report two additional rare cases of EGC with liver metastasis. PMID- 8231061 TI - Early gastric cancer with juxta lymph node (n3) metastasis. AB - From 1976 to 1992, a total of 714 cases of early gastric cancer (EGC) were treated by resection in the First Department of Surgery, Kurume University Hospital. For EGC, the overall lymph node metastasis rate is generally 10%, with the lymph node metastasis observed mainly in the Group 1 perigastric nodes in both mucosal cancer and submucosal cancer. Three rare cases of an early gastric cancer with Group 3 juxta regional lymph node metastasis are described because Group 3 lymph nodes are rarely discovered. To our knowledge, only 14 other cases of EGC with juxta regional lymph node metastasis have been reported in the Japanese literature. PMID- 8231062 TI - Strumal carcinoid of the ovary. AB - A strumal carcinoid arising in a benign cystic teratoma of the left ovary was reported in a 41 year-old woman. The solid tumor was histologically a trabecular carcinoid tumor associated intimately with thyroid follicle-like structures. By Grimelius staining, argyrophil granules were found in the cytoplasm of the tumor cells. The final diagnosis of strumal carcinoid, however, was established by the confirmation of thyroid tissue. PMID- 8231063 TI - Retrospective studies of synchronous double early gastric cancer. AB - From 1984 to 1990, 922 cases of gastric cancer underwent resection in the First Department of Surgery, at Kurume University Hospital. Of these, 381 (41.3%) cases were early gastric cancer (EGC). Among these EGC, synchronous double early gastric cancer (double EGC), according to the diagnostic criteria of Moertel, was found in 49 (12.9%) cases involving 98 foci. In cases of a double EGC, the primary and secondary foci can be distinguished. Double EGC occurred more frequently in males than females (7.1:1). In the elderly, the incidence of a double EGC was higher than the incidence of a single EGC. Double EGC occurred more frequently in the lower third of the stomach than a single EGC. And a macroscopically elevated and histologically differentiated type primary focus was more frequently found in double EGC than in single EGC. Of the 49 secondary foci in the double EGC, 12 (24.5%) foci were a microcarcinoma of less than 5 mm in greatest diameter, and 48 (98%) were limited to the mucosa (m-cancer). Therefore, a preoperative detailed examination of the whole stomach by endoscopy and an intraoperative careful search under gastrotomy must be made in the remnant of the stomach in cases of distal or proximal partial gastrectomy to ensure adequate excision. PMID- 8231064 TI - Lymphocytic alveolitis associated with asymptomatic primary biliary cirrhosis. PMID- 8231065 TI - Low-dose/long-term erythromycin for treatment of bronchiolitis obliterans organizing pneumonia (BOOP). AB - Bronchiolitis obliterans organizing pneumonia (BOOP) is a pathologic entity characterized by intraluminal fibrosis of distal air spaces. Corticosteroids have been widely used for the treatment of this condition, and most patients showed a dramatic response to it. However, long-term treatment with corticosteroids, which often increases the risk of several undesirable side-effects, is usually required because a relapse tends to occur soon after termination of treatment. We administered erythromycin (EM) at low-dose (600 mg daily) for 3-4 months to 6 patients with BOOP, and obtained a good clinical, radiological, and physiological improvement. This suggests that EM can be successfully used, instead of corticosteroids, in the treatment of BOOP. PMID- 8231066 TI - Evaluation of the intramembrane particle count in biopsied myocardium from patients with idiopathic cardiomyopathy. AB - To clarify the abnormalities of myocardial membrane structure in cardiomyopathy, the density of intramembranous particles (IMP) of mitochondria, sarcoplasmic reticulum and sarcolemma, and the number of caveolar necks was evaluated by freeze-fracture morphology. The subjects consisted of 17 patients with hypertrophic cardiomyopathy (HCM), 6 patients with dilated cardiomyopathy (DCM), and 7 patients as controls. The results showed that the numbers of IMPs on the membrane faces of mitochondria, sarcoplasmic reticulum and sarcolemma were significantly decreased in HCM and DCM, and that they were decreased more markedly in DCM than in HCM. The number of caveolar necks was significantly decreased in DCM. Thus, the decreased numbers of IMPs on myocardial membranes and the decrease in caveolar necks may be important characteristics resulting from abnormal myocardial membrane metabolism in cardiomyopathies, HCM and DCM. PMID- 8231067 TI - Clinicopathological studies of multiple cancers in the large bowel and other organs. AB - During the past 17 years, 1,463 cases of cancer of large intestine, there were 117 cases (8.0%) with overlapping cancer in other organs. The male/female ratio was 1.0:1.1 and the mean age was 65.8 years old. The region of existence in other organs was in the order of frequency: stomach 28.3%, uterus 26.3%, and liver 8.1%. The location and histology showed almost similar results to that of single cancer. In comparison with single cancer, overlapping cancer cases showed a high complication rate of adenoma. So it was suggested that a region of cancer development was present but latent. The preceding cases of cancer of the uterus almost all underwent radiotherapy. About 36% of their histology of colon cancer were mucinous carcinoma suggesting at least the development of cancer by radiation. PMID- 8231068 TI - Experimental transmission of HTLV-I into C3H/HeJ mice. PMID- 8231069 TI - [The dosage creates the poison]. PMID- 8231070 TI - [Burnout and suicide in nursing personnel]. PMID- 8231071 TI - [Infections in implants]. PMID- 8231072 TI - [Drug induced liver injuries]. PMID- 8231074 TI - [A long way from tribal ethics to the ethics of general human rights]. PMID- 8231073 TI - [Report of experiences with the use of the Pegasus Airwave System in a surgical intensive care unit]. PMID- 8231075 TI - [Psychological aspects of cigarette smoking]. PMID- 8231076 TI - [Nosocomial hospital infections with Staphylococcus aureus]. PMID- 8231077 TI - [Working with cytostatic agents--what has to be watched for? 2]. PMID- 8231078 TI - [Diagnosis and therapy of infections in neutropenic patients]. PMID- 8231079 TI - [New transplantation legislation being prepared. Medical problems at the borderline between life and death]. PMID- 8231080 TI - [Answers to the questions about the reason and meaning of suffering]. PMID- 8231081 TI - Effects of epinephrine, phenoxybenzamine, and propranolol on maximal exercise in sheep. PMID- 8231082 TI - Enterotropic coronavirus (mouse hepatitis virus) in mice: influence of host age and strain on infection and disease. AB - The course of enterotropic mouse hepatitis virus (MHV) infection was examined in genetically susceptible (BALB) and resistant (SJL) mice of different ages at inoculation (1, 3, and 12 weeks) and at sequential intervals (1, 2, 3, 5, 10, 20, and 30 days) after oral inoculation with the Y strain of MHV (MHV-Y). Virus was quantified in stomach, upper and lower segments of small intestine, cecum, upper and lower segments of colon, Peyer's patches, mesenteric lymph node, and feces, and tissues were examined microscopically. An infant mouse bioassay was used to quantify virus in all tissues of 3-week-old BALB mice and ascending colons of other mouse groups. MHV-specific serum IgG antibody titers were measured with an enzyme immunoassay, using MHV-S-infected 17 Cl 1 cells as antigen. Lesions were first detectable at 2 days and were most severe in 1-week-old mice and more severe in BALB mice, compared with SJL mice of the same age. Additional BALB mice inoculated at the age of 24 hours developed severe necrotizing enterocolitis, whereas SJL mice inoculated at the age of 24 hours developed lesions equivalent to those in 1-week-old BALB mice. Virus was first detectable at 2 days and virus titers were highest at 2, 3, and 5 days, then diminished on days 10, 20, and 30. Low titers of virus were found in a few mice of different ages and genotypes through day 30. Lesions were most severe and virus titers highest in the ascending colon.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231083 TI - Detection of mouse hepatitis virus by the polymerase chain reaction and its application to the rapid diagnosis of infection. AB - Eight different strains of mouse hepatitis virus (MHV) were analyzed by the polymerase chain reaction (PCR) to see whether two sets of oligonucleotides, which were synthesized based on the published nucleotide sequence of MHV-JHM mRNAs 6 and 7, could be used as universal primers for amplification. Total RNA extracted from virus-infected cells or virus-infected culture fluids was transcribed into cDNA by using reverse transcriptase and oligo(dT) as primer, then the cDNA transcripts were amplified by PCR. The MHV-specific fragments of 199-bp and 241-bp were obtained from all eight strains irrespective of nucleotide differences in the primer regions. The same fragments were also amplified from RNA derived from the liver and brain of MHV-JHM-infected mice as soon as day 1 after intraperitoneal injection, even from the liver from which the virus was not detected. Results of PCR amplification from the liver RNA extracts became positive when more than 10(-2)PFU of MHV-JHM was contained in the PCR reaction mixture. In contrast, anti-MHV antibody was not detected by enzyme-linked immunosorbent assay until day 6 after inoculation. These results suggest that PCR is a very sensitive method to identify a variety of MHV infections in laboratory animals, especially at the early phase of infection. PMID- 8231084 TI - Experimental murine cytomegalovirus infection in severe combined immunodeficient mice. AB - Severe combined immunodeficient (scid) and BALB/c mice were experimentally infected with murine cytomegalovirus (MCMV). Scid mice infected by the intraperitoneal route died or were moribund at dose-dependent times ranging from 12 to 13 days after inoculation for > or = 3.00 x 10(4) plaque forming units (pfu) of virus to 25 days for 1.17 x 10(2) pfu. Histologic lesions included severe adrenal necrosis at low doses and splenic necrosis at high doses. Multinucleate hepatocytes with multiple inclusion bodies were observed at all doses. In visceral organs, the inflammatory response consisted of cell necrosis and neutrophil infiltration. Scid mice infected with 1.00 x 10(3) pfu by the intranasal route were moribund by 24 or 25 days after inoculation. Viral titers in tissues examined from these mice increased in all organs examined until they became moribund. BALB/c mice infected intranasally had detectable virus titers in the adrenal glands, salivary glands, lungs, and spleen by 7 or 14 days after inoculation, but decreased thereafter. These mice remained clinically normal through the infection. In BALB/c mice, histologic lesions were present only in the salivary glands. PMID- 8231085 TI - Contamination of transplantable tumors, cell lines, and monoclonal antibodies with rodent viruses. AB - Different biological materials were tested for murine viral contamination by using the mouse/rat antibody production test. Of 297 tumors examined, 75 (25.3%) were contaminated. Considerable differences in the contamination rate became evident when transplantable tumors from in vitro and from in vivo passages were compared. Of 186 tumors that had been propagated in animals, 36.6% were positive, whereas only 7 of 111 (6.3%) tumors propagated in vitro were contaminated. The highest rate of contamination was detected in mouse tumors. Testing of 135 specimens of mouse origin revealed 46.7% were contaminated, and 57 (70.4%) of 81 samples propagated in mice were positive for murine viruses. Moreover, 6.7% of 90 human tumors that had been passaged in athymic nude mice and 3.5% of 57 rat tumors were positive. Lymphocytic choriomeningitis virus was detected in 4 of 14 hamster tumors. The most frequent contaminant was lactic dehydrogenase elevating virus followed by reovirus 3, lymphocytic choriomeningitis virus, minute virus of mice, mouse hepatitis virus, rat coronaviruses, Kilham rat virus, and Mycoplasma pulmonis. Contamination with reovirus 3 and minute virus of mice was found in 4 (3.7%) of 109 cell lines tested, and 2 of 60 monoclonal antibody preparations or hybridoma cells contained lactic dehydrogenase virus. Contamination with two pathogens was detected in four mouse tumors and in one cell line. PMID- 8231086 TI - Induction of diabetes mellitus in Syrian golden hamsters using stored equilibrium solutions of streptozotocin. AB - Streptozotocin (STZ), a compound composed of a mixture of alpha and beta anomers, is used experimentally for the chemical induction of diabetes mellitus in numerous animal species. It is routinely recommended that STZ be administered rapidly following dissolution because of its alleged instability in solution. In the present study, we examined the effect of varying the length of time from dissolution to administration on the ability of STZ to induce diabetes mellitus in male Syrian golden hamsters and examined STZ stability and state of equilibrium by high-pressure liquid chromatography. Effective diabetes induction was determined by monitoring plasma glucose concentrations 2 and 9 days after STZ treatment. Diabetes was successfully induced with solutions of STZ (50 mg/kg body weight given intraperitoneally on three consecutive days) used either immediately (24 degrees C), 2 hours (24 degrees C), or 5 to 7 days (6 degrees C) after dissolution in 0.1 M acetate buffer at pH 4.4 (storage temperature indicated in parentheses). Mean plasma glucose concentration was significantly higher in all STZ treatment groups at both time points when compared with acetate buffer treated controls. There was no significant difference in plasma glucose concentration between STZ treatment groups. High-pressure liquid chromatography analysis demonstrated that the alpha- to beta-anomeric ratio of STZ had reached equilibrium in 84 minutes at 24 degrees C and by 26 hours at 6 degrees C following dissolution. Recovery of STZ was greater in solutions stored at 6 degrees C than at room temperature (24 degrees C).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231087 TI - Weight gain among juvenile rhesus macaques: a comparison of enriched and control groups. AB - Environmental enrichment techniques for captive primates are aimed at improving their psychological well-being. While behavioral variables are used to measure changes in psychological well-being, physiologic measures (e.g., heart rate, cortisol response) are sometimes gathered in addition to the behavioral evidence. Some of these physiologic indices measure acute changes in the animals' well being, limiting their usefulness. Body weight, however, is a measure of physical well-being that may have meaning as a long-term indicator of psychological well being. We therefore collected body weight data from two groups of rhesus macaques (Macaca mulatta group 1: n = 34, group 2: n = 30) every 8 weeks beginning at the age of 1 year, as they passed through various housing conditions as part of a program to develop a specific pathogen-free breeding colony. One-half of the subjects in each group received a variety of environmental enhancements during all housing conditions; the other half received no enrichment and served as controls. At the beginning of the study (age 1 year), control and enriched subjects did not differ in body weight. Among group-1 subjects, enriched animals weighed significantly more than controls after 4 months of enrichment, and the weight difference was maintained 24 months later. Enriched animals in group 2 never differed in weight from their controls. The order in which different types of enrichment were presented and the extra-cage environment of the two groups differed, which may account for this discrepancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231088 TI - Hematologic and biochemical profiles of selectively bred WHHL rabbits. AB - In Watanabe heritable hyperlipidemic (WHHL) rabbits, hypercholesterolemia and hypertriglyceridemia develop from birth, because of a deficiency of low-density lipoprotein receptors, and are followed by a consequent early development of aortic atherosclerosis. This closely resembles human familial hypercholesterolemia. Starting in 1984, we have developed a closed colony by breeding two male and two female homozygous WHHL rabbits, obtained from Japan (Dr. Watanabe, Kobe University). In our facility, the application of a selective breeding program, strictly based on mating parents that both have high serum lipid concentrations, has produced markedly elevated cholesterol (701 +/- 172 mg/dl, mean +/- SD) and triglyceride (780 +/- 325 mg/dl) concentrations in weaning rabbits. Clinical chemical analysis revealed no kidney or liver function abnormalities even in animals with extremely high lipid concentrations, and hematologic profiles were very similar in WHHL and age-matched New Zealand White rabbits, with the exception of platelet count, which was significantly higher in WHHL rabbits. Platelet aggregation induced by collagen and platelet-activating factor was significantly reduced in WHHL rabbits, whereas thrombin and prothrombin times appeared normal when compared with those in New Zealand White rabbits. PMID- 8231089 TI - Propofol in rabbits. 1. Determination of an induction dose. AB - Propofol was evaluated as an intravenous agent for induction of anesthesia in 94 non-premedicated New Zealand White rabbits (mean body weight 3.25 kg). Lack of chewing oral introduction of an endotracheal tube was chosen as the end-point in signaling loss of consciousness. A rapid and excitement-free loss of consciousness was observed. Using probit analysis, a 50% effective dose of 6.44 mg/kg was calculated. No statistically significant difference between genders was shown. Independent of the dose administered, recovery was rapid and smooth. PMID- 8231090 TI - Propofol in rabbits. 2. Long-term anesthesia. AB - The cardiopulmonary effects of propofol as the sole anesthetic agent administered as a continuous intravenous infusion over 8 hours were evaluated in ten ventilated New Zealand White rabbits. Heart rate, mean blood pressure, arterial oxygen and carbon dioxide tensions, arterial pH, hemoglobin's saturation with oxygen, packed cell volume, and temperature were continuously monitored and recorded at 0 (baseline), 30, 60, 120, 180, 240, 300, 360, 420, and 480 minutes and then hourly for up to 4 hours during recovery. Only light planes of anesthesia could be achieved with a mean propofol infusion rate of 0.876 mg/kg/min. Hypotension and hypoxemia were common features. Four rabbits did not survive the study period and recovery was slow in the surviving animals. With monitoring being the same, Intralipid 10%, the vehicle of propofol, infused into two rabbits (controls) had no adverse effects. Intravenous administration of propofol alone cannot be recommended for long-term anesthesia in the rabbit. Oxygen supplementation appears to be an important element in improving quality of anesthesia and outcome. PMID- 8231091 TI - A simplified approach to the anesthesia of porcine laparoscopic surgical subjects. AB - At a series of laparoscopic surgical workshops, 155 pigs were successfully anesthetised for up to 4 hours by using Profolol (Diprivan) as the anesthetic agent and without provision of positive pressure ventilation. On the basis of our findings, we believe this methodology presents a useful alternative to gaseous anesthesia and provides definite logistical and technical advantages. PMID- 8231092 TI - A technique for catheterization of ferrets for chronic intratracheal material administration. AB - A method was needed for repeated administration of a liquid surfactant suspension into the lungs of nonrodent animals for the purpose of safety testing. Catheters were implanted surgically into the tracheas of 66 juvenile ferrets and exteriorized at the back of the neck. After the ferrets had recovered from surgery, liquid was administered via the catheters into the ferrets' tracheas multiple times, for periods up to 1 month. Ferrets were conscious and free breathing during the intratracheal administration of the liquid. The ferrets tolerated the surgery, catheterizations, and dosings well, with few complications. Most ferrets had an occasional nonproductive reflex cough due to tracheal stimulation by the catheter but continued to gain weight, play, and otherwise behave normally. The catheters of two ferrets became displaced from the trachea into the subcutaneous space. Two other ferrets died acutely during administration of the liquid into their tracheas, as a result of airway obstruction. Relative to alternative methods, however, this procedure provides a safe and simple chronic nonrodent model for safety testing of materials introduced into the lungs. PMID- 8231093 TI - Diagnostic exercise: progressive neurologic disease in a dog. PMID- 8231094 TI - Diagnostic exercise: alopecia on a sheep's fetlock. PMID- 8231095 TI - Aeromonas hydrophila peritonitis in a cotton-top tamarin (Saguinus oedipus), and retrospective study of infections in seven primate species. PMID- 8231096 TI - Studies on the eradication of intestinal protozoa of Syrian hamsters in quarantine and their transfaunation to mice. PMID- 8231097 TI - Enteric coccidiosis in a ferret. PMID- 8231098 TI - Posterior paresis induced by Freund's adjuvant in guinea pigs. PMID- 8231099 TI - Granulocytic leukemia and uterine adenocarcinoma in a white-footed mouse (Peromyscus leucopus). PMID- 8231100 TI - An ataxic mutant rat with dilute coat color. PMID- 8231101 TI - Localized pericardial effusion and right-sided heart tamponade: complications of cardiac surgery in a Hanford miniature pig. PMID- 8231102 TI - Evaluation of inanimate objects on commonly monitored variables in preclinical safety studies for mice and rats. PMID- 8231103 TI - Experimentally induced mortality following repeated measurement of rectal temperature in mice. PMID- 8231104 TI - Use of gonadotropic releasing hormone for ovulating the rabbit model. PMID- 8231105 TI - Extracellular matrix and growth factors: an integrated interplay controlling tissue repair and progression to disease. PMID- 8231106 TI - Lipoprotein(a) and its role in atherothrombotic disease. PMID- 8231107 TI - Transforming growth factor-beta 1 induces collagen IV gene expression in NIH-3T3 cells. AB - BACKGROUND: While recent studies have implicated transforming growth factor-beta 1 (TGF-beta 1) in the development of glomerular scarring, extraglomerular matrix production is frequently associated with glomerulonephritis and is an important determinant of disease progression. TGF-beta 1 may be an important mediator of extracellular matrix synthesis, both by glomerular and extraglomerular mesenchymal cells. TGF-beta 1-mediated collagen IV gene expression was studied in two mesenchymal cell lines. Initial studies were performed utilizing NIH-3T3 cells, a fibroblast-like line derived from murine embryo that has been used to study regulation of fibrillar collagen (collagen I and collagen III) synthesis by TGF-beta 1. Additional studies were performed using normal rat kidney cells (NRK 49F). EXPERIMENTAL DESIGN: Cells were grown in medium supplemented with 0.5% calf serum for 24 hours before treatment with TGF-beta 1. RNA was isolated after the addition of varying amounts of TGF-beta 1 to the cells in culture for varying periods of time, and collagen alpha 1(IV) RNA was quantitated by filter hybridization. Transcription of the alpha 1(IV) and alpha 2(IV) collagen genes was assessed by an in vitro transcription assay. Deposition of collagen IV was identified by immunoblotting. RESULTS: Induction of alpha 1(IV) gene expression by NIH-3T3 cells and by NRK-49F cells was first seen 2 to 4 hours after TGF-beta 1 treatment, and was maximal after 12 to 18 hours. Maximal induction was observed following addition of 5 ng/ml TGF-beta 1 to NIH-3T3 cells, and following addition of 10 ng/ml of TGF-beta 1 to NRK-49F cells. In the presence of cycloheximide, TGF beta 1 induction of alpha 1(IV) mRNA was markedly attenuated in both cell lines, suggesting that this effect of TGF-beta 1 requires protein synthesis. TGF-beta 1 increased transcription of both the alpha 1(IV) and alpha 2(IV) collagen genes by NIH-3T3 cells. CONCLUSIONS: TGF-beta 1 induces collagen IV gene expression in both NIH-3T3 cells and normal rat kidney fibroblasts (NRK-49F cells). Further studies of cytokine-mediated transcriptional regulation of collagen IV, utilizing these cell lines, may provide important information regarding the role of extraglomerular matrix production in the progression of renal disease. PMID- 8231108 TI - Pulmonary granuloma formation in the rat is partially dependent on monocyte chemoattractant protein 1. AB - BACKGROUND: We have examined the role of MCP-1 (monocyte chemoattractant protein 1; also known as monocyte chemotactic and activating factor or the murine JE gene product) in the pathogenesis of glucan-induced granulomatous vasculitis in the rat. While in vitro studies indicate that MCP-1 possesses monocyte chemotactic and activating activities, little is known about its biologic role in pathologic processes. Glucan-induced pulmonary granulomatous vasculitis is an ideal model in which to study the role of MCP-1, because the granulomas develop rapidly and synchronously and are monocyte/macrophage-rich. EXPERIMENTAL DESIGN: The purpose of this study was to define the topographic distribution and temporal pattern of MCP-1 expression in the lungs of rats with evolving glucan-induced granulomatous vasculitis and to determine the effect of neutralization of MCP-1 activity on granuloma formation. Glucan-induced pulmonary granulomatous vasculitis was induced in rats by the intravenous infusion of yeast cell wall glucan. At the indicated time points after glucan infusion, rats were sacrificed and the lungs processed for Northern, immunohistochemical and in situ hybridization analyses of MCP-1 production. Morphometric analysis was used to quantify the effect of neutralization of MCP-1 activity on granuloma formation. RESULTS: Granuloma formation was accompanied by a biphasic increase in steady-state whole lung MCP-1 mRNA levels that peaked at 1 and 6 to 24 hours. In situ hybridization and immunohistochemical analyses revealed that components of the bronchial and vascular walls are responsible for the early rise (1 hour) in MCP-1 mRNA and protein expression, whereas granuloma-associated alveolar macrophages are the predominant source of MCP-1 later (6 to 24 hours) in the evolution of these lesions. Intravenous infusion and/or intratracheal instillation of neutralizing concentrations of anti-rat MCP-1 antibody raised against recombinant rat MCP-1 resulted in a dramatic decrease in the number and size of glucan-induced granulomas as well as in the numbers of mononuclear phagocytes retrieved in bronchoalveolar lavage fluid. CONCLUSIONS: These studies demonstrate that glucan induced granulomatous vasculitis is accompanied by increased local expression of MCP-1 mRNA and protein, that there is a coordinated production of MCP-1 by different cell types within the lung during evolving glucan-induced pulmonary vasculitis, and that MCP-1 plays a requisite role in pulmonary granuloma formation. PMID- 8231109 TI - Temporal and spatial immunolocalization of cytokines in murine chronic granulomatous tissue. Implications for their role in tissue development and repair processes. AB - BACKGROUND: Cytokines have profound effects on various aspects of granulomatous tissue formation. However, there is little information regarding their distribution during tissue development. This study investigated the temporal and spatial distribution of transforming growth factor-beta (TGF-beta), platelet derived growth factor (PDGF), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1) and IL-1 beta in developing granulomatous tissue. EXPERIMENTAL DESIGN: Murine chronic granulomatous air pouches were induced and full thickness biopsies taken at intervals up to 28 days. Samples were prepared for immunohistochemistry and labeled using antibodies against TGF-beta, bFGF, PDGF, EGF, TNF-alpha, IL-1 alpha and IL-1 beta. RESULTS: Immunoreactivity to TGF-beta, PDGF, TNF-alpha, IL-1 alpha and IL-1 beta was localized to a proportion of macrophages within the granulomatous tissue. Immunopositive macrophage numbers increased with time, and with the exception of PDGF were associated with areas of fibrogenesis between days 14 to 28. Heterogeneous labeling of capillaries for EGF was observed within the granulomatous tissue juxtaposed to dermal musculature. Diffuse labeling of bFGF, associated with extracellular matrix, was always observed. After day 14, bFGF immunoreactivity was discretely localized to endothelial cells and the basement membrane of vessels within the granulomatous tissue. TGF-beta immunoreactivity was also associated with extracellular matrix components, being most intense in the area of fibrogenesis between 14 and 28 days. Occasional fibroblasts were also labeled with TGF-beta in this region. CONCLUSIONS: The spatial and temporal confinement of the individual cytokines suggests that a sequential coordinated process of repair and fibrosis is occurring. It is hoped that these observations will provide a more effective therapeutic approach for the sequential application of cytokines in abnormalities of wound healing. PMID- 8231110 TI - Exocrinopathy resembling Sjogren's syndrome induced by a murine retrovirus. AB - BACKGROUND: Sjogren's syndrome (SS) is characterized by lymphocytic infiltration into, and destruction of exocrine glands, resulting in dryness of the mouth and eyes. The disease is considered to have an autoimmune etiology, however, its etiopathogenesis remains largely unknown. Recently, retrovirus is suggested to participate in the pathogenesis of SS, because SS-like lesions are reported in HIV infection or in human T cell leukemia virus type I infection. Moreover, human intracisternal A-type retroviral particles are reported to be detected in SS patients. During the course of our study on the histopathology of mice infected with a murine retrovirus, we happened to find SS-like exocrinopathy in those mice. EXPERIMENTAL DESIGN: Four-week-old C57BL/6 (B6) mice were injected intraperitoneally with LP-BM5 murine leukemia virus. This virus is known to induce splenomegaly, lymphadenopathy followed by lymphoid malignancy, and profound immunodeficiency in sensitive strains of mice. From 4 to 16 weeks after the virus inoculation, the infected mice were sacrificed and their submandibular and lacrimal glands were analyzed light and electron microscopically and immunohistochemically. The existence of the virus in the lesion in situ was also analyzed by the same method, and additionally by a polymerase chain reaction method. RESULTS: Periductal lymphocytic infiltration into the submandibular and lacrimal glands was observed in all the virus-infected mice at 4 weeks after the infection and progressed with time. Extraglandular lymphocytic infiltration was also observed in liver, kidney, lung, and pancreas. Immunohistochemical examination revealed that most infiltrating cells into the glands were composed of CD3+ T cells (CD4-dominant), Mac-1+ cells, and B220+ cells. The virus genome was detected in submandibular glands by immunohistochemistry or by polymerase chain reaction. In addition, retroviral particles were secreted into the lumen of exocrine ducts of submandibular glands. CONCLUSIONS: This might be an SS animal model that is induced by a certain defined retrovirus. This experimental system might provide us with valuable information for analyzing the mechanisms of how a retrovirus could induce SS. PMID- 8231111 TI - Alterations in neurofilament mRNA in hereditary canine spinal muscular atrophy. AB - BACKGROUND: Hereditary canine spinal muscular atrophy (HCSMA) is a dominantly inherited motor neuron disease in which distal axonal caliber is reduced in lower motor neurons. Because several animal models show that neurofilament protein gene expression is a major determinant of axonal caliber, we began an examination of neurofilament gene expression in HCSMA early in the clinical disease to determine whether this family of proteins was selectively affected and could thus possibly contribute to the morphologic and functional alterations characteristic of the disease. EXPERIMENTAL DESIGN: We used quantitative in situ hybridization to compare levels of mRNA encoding neurofilament protein subunits in lateral ventral horn neurons from the cervical spinal cord enlargement (C7-C8) in 10-week-old homozygous HCSMA and control dogs. Each slide contained a spinal cord section from a control and an HCSMA dog in order to make within-slide comparisons. The mean number of grains/neuron and the mean neuronal grain density for the HCSMA section were divided by that value for the control section on each slide. The means of these ratios for each mRNA species (i.e., neurofilament subunits and total polyadenylated mRNA (poly-A+) were then compared statistically. RESULTS: The levels of mRNA encoding the low molecular weight neurofilament protein subunit were significantly different from levels of mRNA encoding the high molecular weight neurofilament protein subunit and poly-A+ mRNA in dogs with HCSMA compared with control dogs. The neuronal levels of poly-A+ mRNA were comparable in dogs with HCSMA and controls. CONCLUSIONS: If neurofilament protein subunit levels are found to follow the mRNA levels in this animal model, our results would suggest that decreased expression of the low molecular weight neurofilament gene is sufficient to inhibit neurofilament function, i.e., maintenance of axonal caliber, probably by disrupting normal neurofilament assembly. PMID- 8231112 TI - Induction of perlecan gene expression precedes amyloid formation during experimental murine AA amyloidogenesis. AB - BACKGROUND: In a murine model of AA amyloidosis, it has been demonstrated that perlecan, the basement membrane heparan sulfate proteoglycan, is co-deposited with AA amyloid as it forms in various tissues. The objectives of this study were to determine whether the accumulation of perlecan during amyloidogenesis is associated with induction of perlecan gene expression and, if so, to define the temporal relationship of this induction to the onset of amyloid formation. EXPERIMENTAL DESIGN: Accelerated splenic AA amyloidosis was stimulated in mice by concomitant administration of subcutaneous silver nitrate as an inflammatory stimulus and amyloid-enhancing factor. A kinetic analysis of splenic perlecan mRNA levels during amyloid formation in the spleen was conducted using a reverse transcription-polymerase chain reaction assay. Amyloid deposits were detected histochemically with the Congo red stain and by immunohistochemistry using anti AA antisera. RESULTS: Perlecan mRNA levels increased significantly during amyloidogenesis, increasing 4.1-fold within 72 hours of the amyloidogenic stimulus and subsequently falling to steady-state levels. A 2.0-fold induction of perlecan mRNA occurred by 24 hours post-stimulation, a time at which amyloid was not detectable by either histochemistry or immunohistochemistry. In contrast, control animals administered either the inflammatory stimulus or AEF alone showed no significant change in perlecan mRNA levels. CONCLUSIONS: Increased perlecan mRNA levels account, at least in part, for the accumulation of perlecan in murine splenic AA amyloid deposits. This induction of perlecan gene expression occurs before the onset of amyloid formation, supporting a role for perlecan in the earliest stages of amyloid fibrillogenesis. PMID- 8231113 TI - Alpha-3-fucosyltransferases and their glycoconjugate antigen products in the developing human kidney. AB - BACKGROUND: Three patterns of alpha-3-fucosyltransferase activity have been described in human adult tissues with different acceptor specificity: myeloid, plasma and Lewis. Five- to ten-week embryos express the myeloid enzyme in all tissues tested, then this enzyme is replaced by plasma or Lewis enzymes, with the exception of leukocytes that continue to express the myeloid form of the enzyme in the adult. These enzymes have not been studied as yet in the developing human kidney. EXPERIMENTAL DESIGN: The three different alpha-3-fucosyltransferases were studied in homogenates of mesonephros and metanephros with synthetic oligosaccharide acceptors. The oligosaccharide precursors and products of these enzymes (precursor, H, Le(a), sialyl-Le(a) and Le(b) for type 1 and precursor, H, Le(x), sialyl-Le(x) and Le(y) for type 2) were localized by immunofluorescence with specific antibodies. RESULTS: Only the myeloid alpha-3-fucosyltransferase is detected at 5 weeks in mesonephros and it disappears at 8 weeks. In metanephros, the myeloid enzyme alone is detected between weeks 6 and 8. The plasma enzyme then appears and only at the last trimester of gestation does the Lewis enzyme appear. Three histologic patterns that are concordant with the expression of the alpha-3-fucosyltransferases are observed: I. Inducer, S-shaped body, Bellini and calyce express Le(x) at an early stage when, only the myeloid alpha-3 fucosyltransferase is detected; II. Later, the proximal tubules and descending limbs of Henle's loop express Le(x) (week 9) and sialyl-Le(x) (week 16) when the plasma alpha-3-fucosyltransferase appears; III. Calyceal and collecting systems always express Le(x) and after week 12 Le(a) and Le(b) appear, in accordance with the late appearance of Lewis alpha-3/4-fucosyltransferase. CONCLUSIONS: The sequential appearance of enzymes and their products suggests that during renal organogenesis the myeloid alpha-3-fucosyltransferase is progressively replaced by the plasma enzyme in the proximal tubules and later by the Lewis enzyme in Bellini's ducts and calyce. PMID- 8231114 TI - Coronary vascular injury after transient coronary artery occlusion. AB - BACKGROUND: Microvascular function was examined in dog hearts subjected to transient coronary artery occlusion. EXPERIMENTAL DESIGN: Temporary coronary artery occlusion (15 or 30 minutes) followed by prolonged reperfusion (24 to 72 hours) was performed in dogs. Myocardial blood flow (radioactive microspheres), vascular permeability (human serum albumin uptake), and apparent vascular volume (radioactively labeled erythrocytes) were used as measures of vascular function. Myeloperoxidase was used as a measure of leukocyte uptake. Free radical formation was measured by electron spin resonance using a spin trapping agent (phenyl N tert-butylnitrone). RESULTS: A 30-minute occlusion followed by reperfusion produced prolonged hyperemia, increased vascular permeability, increased apparent vascular volume, and increased leukocyte uptake. Extravasation of red blood cells and carbon black was present within the coronary artery distribution. The altered vascular function was temporally correlated with prolonged leukocyte uptake and free radical production, returning to control values only at 72 hours. A 15 minute occlusion increased vascular permeability at 24 hours, without altering myocardial blood flow or apparent vascular volume. Transmural myocardial leukocyte accumulation after a 15-minute occlusion was minimal in the absence of myocardial infarction or other histologic alterations. Active free radical formation was minimal at 24 hours, with a return of vascular permeability to control values at 48 hours. CONCLUSIONS: The data demonstrate prolonged dysfunction of the coronary microvasculature after a transient 30-minute occlusion of the left anterior descending coronary artery. The prolonged injury is associated temporally with leukocyte uptake and free radical formation in the presence of minimal infarction. A transient 15-minute occlusion produces a different pattern of vascular dysfunction with an abbreviated duration (24 hours), not associated with myocardial necrosis, extensive leukocyte uptake, nor prolonged free radical formation. PMID- 8231115 TI - Assessment of the methods for the detection of Epstein-Barr virus nucleic acids and related gene products in Hodgkin's disease. AB - BACKGROUND: Epstein-Barr virus (EBV) is present in the pathogenic cells of a significant number of cases of Hodgkin's disease, particularly of the mixed cellularity subtype. EBV remains latent and the incidence of detection rate of the genomes and gene products varies greatly with the methods employed. EXPERIMENTAL DESIGN: From a pool of 137 cases of Hodgkin's disease previously studied by cold in situ hybridization (ISH) for the presence or absence of EBV DNA and the immunohistochemical reactivity with anti-latent membrane protein 1 antibody, we selected 24 cases (12 EBV DNA-positive, 12 EBV DNA-negative) for Southern blotting, as well as for study with nonisotopic EBER and BHLF1 oligonucleotide probes and amplification of DNA by polymerase chain reaction. EBV positive cases were further tested with anti-ZEBRA (BZLF1) antibody. RESULTS: The EBV DNA detection rate was found to be lower with Southern blotting compared to ISH and IHC methods because 8 of the 12 EBV positive cases were positive with BamHI W probe and only 7 (of the 8) with XhoI 1.9 kb probe. These 7 cases contained monoclonal episomal circular EBV genomes. All EBV DNA-positive cases showed EBER gene transcription by ISH. EBER probes also reacted with small lymphocytes in all EBV DNA+ and 9 EBV DNA- cases. These EBER-positive small lymphocytes were detected neither by BamHI W DNA probe nor by immunohistochemical methods with anti-latent membrane protein 1 and anti-EBNA2 antibodies. Polymerase chain reaction produced a positive result in all EBV DNA+ cases and 2 (of the 9) EBV DNA- cases containing EBER+ small lymphocytes. This discrepancy was attributed to amplification of EBV in reactive lymphocytes. Anti-ZEBRA antibody was positive in 2 cases (one BHLF1+) suggesting infrequent viral replication and probable abortive lytic cycle. CONCLUSIONS: ISH and immunohistochemical methods are more sensitive than Southern blot for detecting EBV in Hodgkin and Reed Sternberg cells of Hodgkin's disease. Polymerase chain reaction appears to be very sensitive but is less sensitive and specific (amplification of EBV DNA of non-neoplastic lymphocytes) than ISH with non-isotopic EBER oligoprobes, which often detects and localizes EBV in pathogenic cells even when they are in small numbers and also in a few small lymphocytes. In addition, this method offers the advantage of being applied to routinely processed tissue sections. PMID- 8231116 TI - Essential hypertension: historical perspective, mechanisms and an approach to treatment. AB - The mechanisms of essential hypertension have been reviewed and drugs useful in treatment discussed. Although much is known therapy remains largely empiric. Future control of hypertension depends on developments from the laboratory and carefully designed clinical trials that support the empirical. On the horizon, molecular biology promises to be progressing toward unraveling the tangle of the genetic origins of the disease. Perhaps only with this knowledge will completely rational therapy emerge. PMID- 8231117 TI - Management of ventilation tubes: preventing premature extrusion. AB - The Goode T-tube provides a safe and efficient method of ventilating the middle ear space for an extended period of time without random premature extrusion. Each patient needs to be evaluated and followed closely while these tubes are in position. The criteria for the length of time that tubes are retained and the low complication rate are presented. Selective measures for removal of these tubes have provided a reasonable, safe and cost effective means of handling chronic secretory otitis. PMID- 8231118 TI - Continent urinary diversion and bladder substitution. AB - Various forms of supravesical urinary diversion and bladder substitution are currently available. Several factors are considered when deciding which procedure to perform. These include the patient's medical and psychological condition, needs and preferences as well as the surgeon's experience and preferences. While ureterosigmoidostomy is seldom utilized today, it still has a place in properly selected patients with limited life expectancy. Standard ileal conduit remains a reasonable choice in some patients who do not wish or are not suitable candidates for continent forms of diversion. Continent urinary diversion and bladder substitution are attractive choices for most patients needing supravesical urinary diversion. We have found the Charleston pouch to be the ideal form of continent urinary diversion. While continent urinary diversion and bladder substitution improve the quality of life and are well accepted by patients and their urologists, potential complications must be recognized. Long-term follow-up is essential. Finally, patients who have had definitive diversion, such as ileal or colon conduit may now be considered for undiversion or for conversion to continent urinary diversion. PMID- 8231119 TI - Criminalization of health care. PMID- 8231120 TI - The changing image of peer review as relates to the physician. PMID- 8231121 TI - Lacrimae rerum. PMID- 8231122 TI - Eyeglasses and children's schemata. AB - The role of framed eyeglasses in children's person schemata was examined. American first graders (35 girls, 36 boys), rated by their teachers as low, medium, or high in cognitive development, judged peers photographed with and without glasses for physical attractiveness, school performance, conduct, sociability, and sociometric choice. There was a general same-sex bias in the children's ratings. The stimulus persons tended to be rated lower in attractiveness (especially if they were girls), school performance, and conduct when they were wearing glasses than when they were not wearing glasses. Glasses negatively influenced sociometric choice, but only for the most cognitively developed children. These results indicated that adults' stereotypes about people who wear glasses eventually become part of children's person schemata. PMID- 8231123 TI - Gender differences in flashbulb memories elicited by the Clarence Thomas hearings. AB - American students and other adults aged 19-75 completed a questionnaire about flashbulb memories and recollections of autobiographical events elicited by the Senate hearings for confirmation of Clarence Thomas as a Supreme Court Justice. The respondents were less likely to recall vivid image memories than were respondents in earlier studies about memories of the assassinations of John Kennedy, Robert Kennedy, and Martin Luther King. Women were significantly more likely than men to report vivid image memories and recall of autobiographical events elicited by the hearings, but they did not differ significantly from men in the ratings of these memories. Women were also significantly more likely than men to report specific memories of having been victims of sexual harassment and abuse. Women recalled reconsidering incidents in which they might have been victims of sexual harassment more often than men did. Exposure to media coverage did not differ by gender, although the amount of coverage paid attention to did correlate with the number of personal memories elicited. PMID- 8231124 TI - Frightening information and extraneous arousal: changing cognitions and behavior regarding nuclear war. AB - An experiment was designed to demonstrate that fear creates changes, that extraneous arousal modulates the effect of frightening information, and that Janis's (1967) theory may explain these effects. American students watched either a videotape depicting a nuclear attack or a nonfrightening videotape unrelated to nuclear war and were required to perform 10, 20, 35, or 100 jumping jacks. The results of the experiment demonstrated that fear was effective, especially at moderate levels of extraneous arousal, but high levels of arousal eliminated or reversed the effects. PMID- 8231125 TI - The effect of ethnic threat on ethnic self-concept and own-group ratings. AB - The effect of ethnic threat on adolescents' ethnic self-concept and own-group ratings, and the role of self-esteem and ethnic identity in mediating this effect were examined. Hispanic high school students in the United States received either negative (threat) or neutral information about their ethnic group. Ethnic self concept and own-group ratings were then assessed. The negative information resulted in lower overall ethnic group ratings, but not in lower ethnic self concept. Self-esteem was a better predictor of ethnic ratings than ethnic identity was. PMID- 8231126 TI - Gender, ethnicity, and the structure of self-esteem: an attitude theory approach. AB - Consistent with attitude theory, self-esteem was considered in this study to be an attitude toward self, with component self-beliefs that associate or dissociate self with a desired or undesired attribute. It was hypothesized that the structure of self-beliefs underlying self-esteem is different across ethnic and gender boundaries. This hypothesis was tested by measuring the overall self esteem and specific self-belief components in a sample of 643 (305 African American and 338 White) middle school students in two small rural southern towns. In a series of regression analyses within ethnicity by gender groups, the self esteem measure was predicted from indices representing specific self-beliefs associating self with different attributes. Substantial differences were found between the ethnicity by gender groups in the type of beliefs that best predicted self-esteem. PMID- 8231127 TI - Gender, ethnic group, and homelessness as predictors of locus of control among job training participants. AB - The effect of gender, ethnicity, and homelessness on the locus of control orientations of 113 participants in a job training program in the United States was investigated. It was expected that homeless women of color (Black and Hispanic) would have the strongest perception that powerful others influenced their lives. The Black participants tended to believe more strongly in the influence of chance and powerful others than the White participants did. PMID- 8231128 TI - Sex differences in competitiveness and the valuation of money in twenty countries. AB - Sex differences in competitiveness, the valuation of money, and the importance attached to saving money were examined among university students in 20 countries, using questionnaires. In general, the men obtained higher mean scores than the women did. Scores on the trait questionnaires were generally positively intercorrelated, suggesting the presence of a single trait underlying the sex difference. PMID- 8231129 TI - Effects of inequity on job satisfaction and self-evaluation in a national sample of African-American workers. AB - Data from a national survey of African-American workers were used to test predictions from equity theory regarding the "distress" that results from negative inequity (below expected level) and positive inequity (above expected level) in pay and occupational prestige. The results showed differential effects of the two types of inequity on dependent variables of job satisfaction and self report of job-related skill. Although those faced with opposite forms of inequity fostered different strategies for inequity reduction, in each case, workers chose a strategy that was self-protective. PMID- 8231130 TI - Gender and attribution: a reversal of bias? PMID- 8231131 TI - The relationship between South African adolescents' knowledge and fear of AIDS and their attitudes toward people who have AIDS. PMID- 8231132 TI - Batswana children's career aspirations and views of sex roles. PMID- 8231133 TI - The use of robotics technology to study human joint kinematics: a new methodology. AB - Robotics technologies have been modified to control and measure both the force and position of synovial joints for the study of joint kinematics. One such system was developed to perform kinematic testing of a human joint. A 6-axis articulated robotic manipulator with 6 degrees of freedom (DOF) of motion was designed and constructed; a mathematical description for joint force and position was devised; and hardware and software to control forces applied to the joint, as well as position of the joint, were developed. The new methodology was utilized to simulate physiological loading conditions and to perform an anterior-posterior (A-P) translation test on a human cadaveric knee. Testing showed that this new system can stimulate complex loading conditions and also measure the resulting joint kinematics. PMID- 8231134 TI - A method for measuring tension in small ligaments: an application to the ligaments of the wrist carpus. AB - A new technique has been devised for measuring the in situ tension in small ligaments. It is based on measuring the tension in an axially loaded flexible cable with pinned endpoints by deflecting the cable laterally and measuring its lateral load and deformation. Studies were performed in which nylon line and bone ligament bone preparations were placed in a materials tester and loaded in axial tension. Axial load as measured by lateral load and deformation was found to agree with the known load to within 8 percent. The method was sensitive to error in determination of ligament length, nonperpendicularity of the laterally applied load to the long axis of the ligament, and when used in situ, impingement of the ligament on a third bone causing bending. A device, consisting of an LVDT mounted to a rigid frame with its core rod connected to a load cell, was developed. The position of the core rod was controlled by a manual screw drive, and a hook on the other end of the core rod was used to deflect the ligament laterally. This device was applied to the study of tensions in five ligaments of the palmar wrist carpus, in seven cadaver specimens. Results showed that the radioscaphocapitate (RSC) and radiolunate (RL) ligaments had significantly greater tensions than the lunotriquetral (LT), the triquetrocapitate (TC), and scaphocapitate (SC) ligaments. For the four positions of the hand tested, neutral, 14 deg radial and 14 deg ulnar deviation, and 28 deg of extension, ligament tensions were found to be unaffected by position.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231135 TI - Relationship between Achilles tendon mechanical properties and gastrocnemius muscle function. AB - Strain was measured along the length of frog (Rana pipiens) gastrocnemius muscle tendon units (MTU). Maximum muscle tension (P0) was measured, and the MTU was passively loaded to P0. Strain at P0 was measured at eight intervals along the tendon and aponeurosis and was approximately two percent for all regions except the aponeurosis region closest to the muscle fibers where it was about six percent. A computer model predicted sarcomere shortening of up to 0.5 micron due to tendon lengthening which demonstrates that tendons provide a more complex physiological function than simply transmitting muscle force to bones. PMID- 8231136 TI - Biomechanics of ocular pneumoplethysmography. AB - A mathematical analysis of ocular pneumoplethysmography is presented, based on the physiological, anatomical, and biomechanical properties of the eye. Ocular pneumoplethysmography is a clinical procedure involving elevation of intraocular pressure, by application of a suction cup to the sclera, to a level that exceeds ophthalmic artery systolic pressure. As decay in intraocular pressure is allowed, return of retinal artery pulsations indicates ophthalmic artery systolic pressure. We obtain a quantitative relationship between increase in intraocular pressure and applied vacuum, and compare the theoretical predictions with experiments on rabbits in which a variable descending vacuum was applied to bilateral scleral eyecups. The bilateral intraocular pressures were simultaneously recorded from cannulae in the respective vitreous bodies, and the pressures at which return of ocular pulsations were observed were correlated with the scleral vacuums. Regression lines were calculated for three serial determinations in each animal, with two groups of animals distinguished by the inner diameter of the eyecups used. The theoretical results indicate that the relationship between intraocular pressure increase and applied vacuum is independent of Young's modulus, and depends primarily on the ratio of the diameter of the vacuum cup to the diameter of the eye. PMID- 8231137 TI - Multiple minima in inverse hyperthermia temperature estimation problems. AB - Using one-, two-, and three-dimensional numerical simulation models it is shown that multiple minima solutions exist for some inverse hyperthermia temperature estimation problems. This is a new observation that has important implications for all potential applications of these inverse techniques. The general conditions under which these multiple minima occur are shown to be solely due to the existence of symmetries in the bio-heat transfer model used to solve the inverse problem. General rules for determining the number of these global minimum points in the unknown parameter (perfusion) space are obtained for several geometrically symmetric (with respect to the sensor placement and the inverse case blood perfusion model) one-, two-, and three-dimensional problem formulations with multiple perfusion regions when no model mismatch is present. As the amount of this symmetry is successively reduced, all but one of these global minima caused by symmetry become local minima. A general approach for (a) detecting when the inverse algorithm has converged to a local minimum, and (b) for using that knowledge to direct the search algorithm toward the global minimum is presented. A three-dimensional, random perfusion distribution example is given which illustrates the effects of the multiple minima on the performance of a state and parameter estimation algorithm. This algorithm attempts to reconstruct the entire temperature field during simulated hyperthermia treatments based on knowledge of measured temperatures from a limited number of locations. PMID- 8231138 TI - Numerical simulation of the thermal fields occurring in the treatment of malignant tumors by local hyperthermia. AB - Dual beam microwave heating is known to deposit heat at depth in a medium. Thus transient heating times may be reduced and more even heating may be obtained. Such a system has potential in the treatment of cancer by hyperthermia. A theoretical analysis of this situation is presented here. A simulation has been made of the thermal fields generated in the treatment of malignant tumors using local hyperthermia. The simulation utilizes the alternating direction implicit method which is particularly suited to the solution of the governing equations, and provides rapid convergence in multiple dimensions. The simulation is three dimensional in temperature, with variations occurring through two spatial coordinates and one time coordinate. The simulation can accommodate the transient flow of heat due to conductive heat transfer through tissues such as healthy tissue, malignant tumors, cartilage and bone, convective heat transfer through perfusion in the tissue and flow through the arteries, and heat generation from sources such as microwave beams. Small changes in the thermophysical properties of the tissue, and the blood perfusion rates are shown to exhibit only minor effects on the thermal fields, whereas the power of the heat sources, and the conductive flux are shown to have major effects on the thermal fields. The effects of adjacent physiological structures such as arteries and bones have also been determined. The temperature fields have been found to be weakly dependent on the increased perfusion rates encountered in the arteries except when the perfusion rate in the artery exceeds that in the tissue by at least one order of magnitude.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231139 TI - Determination of uterine activity during labor by means of infrared thermography. PMID- 8231140 TI - A semilinear state and parameter estimation algorithm for inverse hyperthermia problems. AB - An improved state and parameter estimation algorithm has been developed which decreases the total computational time required to accurately reconstruct complete hyperthermia temperature fields. Within this improved iterative estimation algorithm, if the change in the unknown perfusion parameters is small a linear approximation scheme is implemented in which the old Jacobian matrix (the sensitivity matrix) is used, instead of recalculating the new Jacobian matrix for the next iteration. In the hyperthermia temperature estimation problem the relationship between the temperature and the blood perfusion based on the bioheat transfer equation is generally nonlinear. However, the temperature can be approximated as a linear function of the blood perfusion over a certain range thus allowing this improved approach to work. Results show that if the temperature is approximated as a linear (or quasi-linear) function of the blood perfusion, the linearizing approach considerably reduces the CPU time required to accurately reconstruct the temperature field. The limiting case of implementing this approach is to calculate the Jacobian matrix for each iteration, which is identical to the approach used in the original nonlinear algorithm. Critical values of determining whether or not there is a need to recalculate the new Jacobian matrix during the iterations are presented for several inverse hyperthermia temperature estimation problems. PMID- 8231141 TI - Viscous airflow through a rigid tube with a compliant lining: a simple model for the air-mucus interaction in pulmonary airways. AB - The respiratory tract of mammals is lined with a layer of mucus, described as viscoelastic semi-solid, above a layer of watery serous fluid. The interaction of these compliant layers with pulmonary airflow plays a major role in lung clearance by two-phase gas-liquid flow and in increased flow resistance in patients with obstructive airway diseases such as cystic fibrosis, chronic bronchitis and asthma. Experiments have shown that such coupled systems of flow compliant-layers are quite susceptible to sudden shear instabilities, leading to formation of relatively large amplitude waves at the interface. Although these waves enhance the lung clearance by mobilizing the secretions, they increase the flow resistance in airways. The objective of this paper is to understand the basic interaction mechanism between the two media better by studying airflow through a rigid pipe that is lined by a compliant layer. The mathematical model that has been developed for this purpose is capable of explaining some of the published experimental observations. Wave instability theory is applied to the coupled air-mucus system to explore the stability of the interface. The results show that the onset flow speed for the initiation of unstable surface waves, and the resulting wavelength, are both very sensitive to mucus thickness. The model predicts that the instabilities initiate in the form of propagating waves for the elastic mucus where the wave speed is about 40 percent of the flow speed. The wavelength and phase speed to air velocity ratio are shown to increase with increasing mucus thickness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231142 TI - Surfactant effects on fluid-elastic instabilities of liquid-lined flexible tubes: a model of airway closure. AB - A theoretical analysis is presented predicting the closure of small airways in the region of the terminal and respiratory bronchioles. The airways are modelled as thin elastic tubes, coated on the inside with a thin viscous liquid lining. This model produces closure by a coupled capillary-elastic instability leading to liquid bridge formation, wall collapse or a combination of both. Nonlinear evolution equations for the film thickness, wall position and surfactant concentration are derived using an extended version of lubrication theory for thin liquid films. The positions of the air-liquid and wall-liquid interfaces and the surfactant concentration are perturbed about uniform states and the stability of these perturbations is examined by solving the governing equations numerically. Solutions show that there is a critical film thickness, dependent on fluid, wall and surfactant properties above which liquid bridges form. The critical film thickness, epsilon c, decreases with increasing mean surface tension or wall compliance. Surfactant increases epsilon c by as much as 60 percent for physiological conditions, consistent with physiological observations. Airway closure occurs more rapidly with increasing film thickness and wall flexibility. The closure time for a surfactant rich interface can be approximately five times greater than an interface free of surfactant. PMID- 8231143 TI - Airway area by acoustic reflection: the two-microphone method. AB - This report deals with noninvasive imaging of airway geometry based upon information contained in acoustic reflections measured at the mouth. Here we describe a new theoretical approach that enables development of a new miniaturized apparatus. Unlike the single-transducer systems used currently, this new strategy is based upon a two-transducer system that is a variant of that suggested originally by Shroeder (1967). We have developed, implemented, and tested computational algorithms necessary to reconstruct airway dimensions from acoustic reflection data using this two-transducer strategy. PMID- 8231144 TI - Distensibility of small pulmonary blood vessels. AB - Although vasomotor activity in small pulmonary vessels has been studied extensively in the past, using the concept of resistance to flow, information on the distensibility of these vessels is very sparse. In an attempt to reduce this deficit, we adapted a theoretical method developed for small systemic vessels, to estimate distensibility of pulmonary resistance vessels in experimental animals and man. Pressure-flow data from 11 dogs and 10 human subjects (5 control subjects and 5 patients with long-standing left heart failure) were used to calculate distensibility of small pulmonary vessels. The conductance, G, was calculated from these data as the ratio of blood flow to driving pressure. The slope of the relationship between the logarithm of G1/4 and the average distending pressure (ADP) provides a graphic picture of circumferential extensibility, E, defined as percent change in radius for an infinitesimal change in ADP. Results indicate that: (1) the value of E in dogs was 1.85 +/- .40 mmHg-1 for the control state, which decreased to 1.45 +/- .43 mmHg-1 during norepinephrine administration; however, the decrease in the value was not statistically significant (p = 0.53); (2) the value of E in control human subjects was 3.38 +/- .47 mmHg-1 and the value of E in patients with left heart failure was -0.64 +/- 0.39 mmHg-1; the difference was significant (P = .0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231145 TI - Laser anemometry measurements of steady flow past aortic valve prostheses. AB - An experimental investigation was conducted in steady flow to examine the fluid dynamics performance of three prosthetic heart valves of 27 mm diameter: Starr Edwards caged ball valve, Bjork-Shiley convexo-concave tilting disk valve, and St. Vincent tilting disk valve. It was found that the pressure loss across the St. Vincent valve is the least and is, in general, about 70 percent of that of the Starr-Edwards valve. The pressure recovery is completed about 4 diameters downstream. The velocity profiles for the ball valve reveal a large single reversed flow region behind the occluder while those for the tilting disks valves reveal two reversed flow regions immediately behind the occluders. Small regions of stasis are also found near the wall in the minor opening of Bjork-Shiley valve and in the major opening of St. Vincent valve. The maximum wall shear stresses of the three valves at a flow rate of 30 l/min are in the range 30-50 dyn/cm2 which can cause hemolysis of attached red blood cells. The corresponding maximum Reynolds normal stresses are in the range of 1600-3100 dyn/cm2. The Reynolds normal stresses decay quickly and return approximately to the upstream undisturbed level at about 4 diameters downstream while the wall shear stresses decay at a slower rate. The maximum Reynolds normal stresses occur at about 1 diameter downstream while the maximum wall shear stress is at about 2 diameters downstream. In general, the St. Vincent valve has better performance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231146 TI - The steady expiratory pressure-flow relation in a model pulmonary bifurcation. AB - Experiments were conducted over a range of Reynolds numbers from 50 to 8000 to study the pressure-flow relationship for a single bifurcation in a multi generation model during steady expiratory flow. Using the energy equation, the measured static pressure drop was decomposed into separate components due to fluid acceleration and viscous energy dissipation. The frictional pressure drop was found to closely approximate that for an equivalent length of curved tube with the same curvature ratio as in the model bifurcation. The sensitivity of these results to changes in airway cross-sectional shape, non-planar configuration, and flow regime (laminar-turbulent) was investigated. In separate experiments using dye visualization and hot-wire anemometry, a transition to turbulent flow was observed at Reynolds numbers between 1000 and 1500. Transition had very little effect on the pressure-flow relation. PMID- 8231147 TI - A computer simulation of the blood flow at the aortic bifurcation with flexible walls. AB - To understand the role of fluid dynamics in atherogenesis, especially the effect of the flexibility of arteries, a two-dimensional numerical model for blood flow at the aortic bifurcation with linear viscoelastic walls is developed. The arbitrary Lagrangian-Eulerian method is adopted to deal with the moving boundary problem. The wall expansion induces flow reversals or eddies during the decelerating systole while the wall contraction restricts them during the diastole. A flexible bifurcation experiences the shear stresses about 10 percent lower than those of a rigid one. PMID- 8231148 TI - Numerical studies of three-dimensional arterial flows in reverse curvature geometry: Part I--Peak flow. AB - A three-dimensional flow simulation at Repeak = 192 and 580 was made in a smooth reverse curvature model that conformed to the gentle "S" shape from a human left femoral artery angiogram. The objective of this numerical investigation was to find the changes in pressure, shear stress, velocity profile, and particle path occurring in the double-curved arterial vessel. Due to the impingement of blood at the outer wall in the first bend region, the wall shear stress approached 40 dyne/cm2--a value over twice as large as in the straight upstream segment. Conversely, at the inner wall in the first bend, a low shear stress region was found where the value of the shear stress was consistently smaller than that in the straight section. The initiation of centrifugal effects caused by the first bend could clearly be seen at Repeak = 580, but due to the close proximity of the reverse curvature segment, the momentum effect due to the second bend overshadowed the centrifugal effect. Hence, only near the end of the second bend did the centrifugal effect due to the second bend result in a double-spiral secondary motion. In addition, the numerically calculated pressure drop data were in agreement with prior experimental values. PMID- 8231149 TI - The orientation of an intramyocardial vessel affects its mechanical loading by the surrounding myocardium. PMID- 8231150 TI - Viscosity sensing with lamb-wave microsensor: dimethylsulfoxide solution viscosity as a function of temperature. AB - Recently, a new microsensor employing low-velocity ultrasonic Lamb waves was developed and demonstrated to be capable of measuring the viscosity of solutions in small volumes. The microsensor, when attached to a temperature-controlled stage, can measure viscosity as a function of temperature. In this investigation, the ultrasonic Lamb-wave oscillator is employed to experimentally measure the viscosity of dimethylsulfoxide (Me2SO) solutions as a function of temperature. The microsensor and the experimental procedure are described and results for 1M, 3M, and 5M Me2SO aqueous solutions are presented. Dimethylsulfoxide is a compound commonly employed as a cryoprotectant in cryopreservation, the low-temperature preservation of biological materials. The temperature dependence of viscosity obtained through this study can be used in determining the probability for ice nucleation in biological materials, a parameter of importance during cryopreservation. PMID- 8231151 TI - Neural network segmentation of magnetic resonance spin echo images of the brain. AB - This paper describes a neural network system to segment magnetic resonance (MR) spin echo images of the brain. Our approach relies on the analysis of MR signal decay and on anatomical knowledge; the system processes two early echoes of a standard multislice sequence. Three main subsystems can be distinguished. The first implements a model of MR signal decay; it synthesizes a four-echo multiecho sequence, in order to add images characterized by long echo-times to the input sequence. The second subsystem exploits a priori anatomical knowledge by producing an image, in which pixels belonging to brain parenchyma are highlighted. Such anatomical information allows the following submodule to distinguish biologically different tissues with similar water content, and hence similar appearance, which might produce misclassifications. The grey levels of the reconstructed sequence and the output of the second module are processed by the third subsystem, which performs the segmentation of the sequence. Each pixel is assigned to one of five different tissue classes that can be revealed with brain MR spin echo imaging. With a suitable encoding, a five-level segmented image can then be produced. The system is based on feed-forward networks trained with the back-propagation algorithm; experiments to assess its performance have been carried out on both simulated and clinical images. PMID- 8231152 TI - PC-based system for an objective quantification of manual movement disability for clinical and scientific purposes. AB - In clinical management and research of movement disorders exact knowledge about the extent of motor impairment is essential. This paper presents a computer program which allows for an objective measurement of manual movement disability. The program was developed for standard hardware and can easily be used in a variety of clinical and research environments. The program runs on MS-DOS computers and uses a Microsoft computer mouse as the only input device. The temporal resolution is 100 Hz, the spatial resolution 400 dots per inch. The user may choose between standard test sets or he may design sets according to his individual needs from a pool of available protocols which includes tracking tasks, ballistic tasks, complex sequential tasks, and finger tapping. All tasks are implemented in a similar way in order to keep the test environment as consistent as possible for the patient. The patient must usually carry out movements which correspond to the movements of a target symbol on the computer screen. This entails the manipulation of a follower symbol, also visible on the computer screen, via the computer mouse. The program itself and the theoretical background of the protocols are described in the paper. Additionally, preliminary results from pilot experiments are presented. PMID- 8231153 TI - Precision and accuracy of bone landmarks in characterizing hand and wrist position. AB - Characterization of the motion of the hand and wrist requires reference to the underlying bones which, for three-dimensional analyses, are assumed to be rigid bodies. Stereoradiogrammetric techniques involving the identification of prominent bone landmarks have been used as the standard against which surface markers used for in vivo testing have been evaluated. The precision and accuracy with which the 3D positions of bone landmarks in the hand and wrist could be determined was evaluated in a small inter-observer and inter-cadaver study and compared to the precision and accuracy with which implanted lead markers could be located. A subset of landmarks suitable for evaluating wrist and metacarpal phalangeal joint motion was identified; the mean precision for identifying these points was better than 1.1 mm in all hand positions with a mean inter-observer accuracy of 2.3 mm. These values show that the average uncertainty in locating bone landmarks is at best roughly twice that for implanted markers. PMID- 8231154 TI - Analysis of all possible combinations of four measurements determining true propagation in arteries. AB - All possible combinations of four measurements of blood pressure, blood flow and vascular diameter are examined by transmission-line theory. It is found that only nine measurement combinations can give the attenuation coefficient gamma, reflection coefficient R and characteristic impedance Z0 simultaneously. At least one pressure measurement must be included. Their general expressions with arbitrary measuring locations are presented, together with some simplified forms which cover all the previous methods. A greater choice of method is thereby made available for use in practice. The results show that, regardless of the measurement locations, all combinations can be solved in the order gamma first, R second and Z0 last. PMID- 8231155 TI - A new geometric description of the right ventricle. AB - An accurate model of the geometry of the right ventricle would be particularly useful for volume determination by imaging techniques, as well as for mechanical modelling. In this study, a new mathematical model for the right ventricle geometry based on its crescentic cross-sectional shape is proposed and used to calculate its volume in imaged ex-vivo casts. An excellent correlation (r = 0.97) was found between volume as derived from the model and actual volume measured by water displacement. PMID- 8231156 TI - An integrated procedure to assess knee-joint kinematics and kinetics during gait using an optoelectric system and standardized X-rays. AB - This paper describes a 3-D gait analysis system, which combines optoelectric motion tracking and a standardized X-ray procedure, to calculate the net knee joint forces and moments of a normal subject group during walking. The optoelectric system collects kinematic data from infra-red LED markers placed at selected skin surface locations and projecting probes attached to the lower limb. A standardized X-ray procedure is used to move surface markers into their designated bony landmarks based on individual bone structure, which reduces the error caused by uncertainty of skin-surface marker locations. Based on moved-in marker information, different joint coordinate systems are proposed for kinematic and kinetic analysis of the knee joint. Normalized data of knee angles, net reaction forces and net moments from 35 young, normal subjects are presented. PMID- 8231157 TI - An adaptive finite-element approach for the analysis of dental restorations. AB - The present paper deals with the problem of stresses generated during the setting process of polymer tooth fillings. A finite element procedure capable of accurately predicting the stresses along an interface between two different materials is presented. Adaptation of the finite element mesh is carried out based on a simple error estimator. The procedure is tested against benchmark problems and then applied to polymer tooth fillings. The results of the study are relevant in understanding and improving the behaviour of dental restorations. PMID- 8231158 TI - A new instrument for the measurement of forces on beds and seats. AB - Knowledge of the interaction of forces between persons and the bed in which they lie or the seat on which they are sitting, provides an insight into the loading of their muscles, bones and soft tissue. To determine the total forces on the body-supporting surfaces (backrest, seat pan, foot rest) resolved in components perpendicular and parallel to these surfaces a new instrument has been developed, with which the forces perpendicular and parallel to three different freely adjustable body-supporting surfaces can be registered. During the first measurements the forces on a bed were measured when a person sits in a bed with the backrest at an angle of 45 degrees to the horizontal and the mattress horizontal. The measurements on a healthy population (mean mass = 77 kg, SD = 11 kg) showed an accuracy of +/- 10N. In this position the mean shear force on the seat pan was 97 N. PMID- 8231159 TI - Quantitative analysis of four EMG amplifiers. AB - Four typical EMG amplifiers were tested quantitatively to observe the diversity and specificity of available equipment. Gain, phase, common mode rejection ratio (CMRR) and noise characteristics were measured for each device. Various gain and phase responses were observed, each best suited to specific application areas. For all amplifiers, the CMRR was shown to decrease dramatically in the presence of input impedance mismatches of more than 10 k omega between the two electrodes. Because such impedance mismatches are common on the skin surface, these results indicate that proper skin preparation is required to maximize the noise rejection capabilities of the tested amplifiers. PMID- 8231160 TI - Development of a concentric water-displacement model lung. AB - Simulation of lung ventilation using a model lung can provide a means of evaluating lung function tests, mathematical models and computer algorithms. We describe a new water-displacement lung model, which can simulate lung volumes up to 3.8 l and tidal volumes up to 1 l. Gas mixing is ensured by using a ring of venturi devices. Model compliance and airways resistance are described. PMID- 8231161 TI - Viscoelastic properties of the human lumbodorsal fascia. AB - The purpose of this study is to provide better understanding of the mechanical response of the lumbodorsal fascia to dynamic and static traction loadings. Since the fascia shows a viscoelastic behaviour, tests in which time is a variable were used, namely hysteresis and stress relaxation. Load-strain and load-time curves obtained from the hysteresis and stress-relaxation tests point out three different phenomena. First, an increase in stiffness is noticed when ligaments are successively stretched, i.e. strains produced by successive and identical loads decrease. Second, if a sufficient resting period is allowed between loadings, stiffening is reversed and strains tend to recover initial values. The third phenomenon, observed in stress-relaxation tests as time progresses, is ligament contraction in stretched and isometrically held samples. This third phenomenon may be explained by the possibility that muscle fibres capable of contracting spontaneously could be present in lumbodorsal fascia ligaments. PMID- 8231162 TI - New method for estimating explosive anaerobic leg power. AB - In this article, a new technique for estimation of explosive anaerobic leg power is proposed. The method relies on accurately measuring the time duration of muscular contraction and the time spent off the ground while performing a vertical jump. Contribution of arms to leg power measurement is minimized by fixing arms at belt line during the test. Results from nine athletes are presented and discussed. A comparison with other methods is also discussed. PMID- 8231163 TI - Seven-channel digital telemetry system for monitoring and direct computer capturing of biological data. AB - A seven-channel telemetry system for collection and display of biological data is presented. The system can amplify bioelectrical signals in the range of 2 microV to 200 mV and has a bandwidth of 0.1-80 Hz. After multiplexing, the signals are digitized with a resolution of 8 bits. The data are frequency modulated directly on a VHF transmitter. After receiving the data on a VHF receiver, they are routed directly to the RS232 input connector on the PC. Thereby the advantage of direct communication between the transmitter and the PC can be utilized. Expensive analog equipment is avoided and display of the signals on the PC screen as well as signal analysis can be performed. The system has been tested and was found to be stable and highly reliable. PMID- 8231164 TI - Altered hepatic production of apolipoproteins B and E in the fasted septic rat: factors in the development of hypertriglyceridemia. AB - The etiology of hypertriglyceridemia associated with sepsis remains unclear, but we will attempt to elucidate its character by studying the hepatic production of apolipoproteins B and E. Male Lewis rats (260-330 g) were assigned to two groups, control (n = 5) and septic (n = 5). The septic group was injected with 2 x 10(8) live Escherichia coli colonies/100 g body wt. Food was removed from all rats after injections. Twenty-four hours later a recirculating in situ liver perfusion was performed for 120 min with KRB buffer, containing L-[35S]methionine. The production of apolipoprotein B (apo B), apolipoprotein E (apo E), albumin, and transferrin was determined by immunoprecipitation. The septic rats showed a protein-specific response to sepsis. The total protein secreted increased throughout each perfusion, septic greater than control. Apo B production was increased 2.6-fold in the septic versus control groups (P = 0.037), while apo E production was decreased by 2.9 times control (P = 0.036). Albumin production was decreased 2-fold in the septic group (P = 0.002). The increased hepatic production of apo B represents an increased number of very low density lipoprotein (VLDL) particles and contributes to the elevated VLDL triglyceride levels seen in sepsis. In contrast, decreased apo E production may result in a diminished ability for peripheral and/or hepatic receptor recognition of VLDL and VLDL remnants, respectively. Each of these changes are factors in the development of hypertriglyceridemia in sepsis. PMID- 8231165 TI - Putative vascular endothelial cell chemotactic factors: comparison in a standardized migration assay. AB - While a number of chemoattractants of vascular endothelial cells have now been identified in vitro, differences in methodology preclude comparisons of substances evaluated in different assays. Here, we report a standardized chemotactic assay in which the migration of calf pulmonary artery endothelial cells in a 48-well microchemotaxis chamber was determined. Nonstimulated (control) migration was remarkably constant (mean +/- SD, 96 +/- 14) from plate to plate, thus allowing the indexing of relative migration of stimulated cells to that of nonstimulated cells in the control wells of that plate. Based on the relative migrations observed in response to each of the substances evaluated, those proving to be stimulatory of migration were placed in rank order by potency. The growth factors epidermal growth factor, transforming growth factor alpha, and basic fibroblast growth factor (followed by pentosan polysulfate, plasmin, fibronectin, fibrinogen, granulocyte-macrophage colony stimulating factor heparin, adenosine, and MgSO4) were the most potent. Only the platelet factors platelet-derived growth factor-BB and platelet activating factor proved inhibitory of migration. Combining fibrinogen with other chemoattractants produced either stimulation or inhibition in comparison to the migration observed with fibrinogen alone, suggesting that more than one signal transduction mechanism was, in all likelihood, invoked by the various agents. This assay will allow the rapid screening and rank ordering of additional putative chemoattractants, will facilitate the study of the biochemical mechanisms involved in endothelial cell migration, and will permit the evaluation of pharmacologic agents capable of modulating stimulated or unstimulated migration. PMID- 8231166 TI - Flow cytometric DNA and cytomorphometric analysis in renal cell carcinoma: its correlation with histopathology and prognosis. AB - Cell material from 49 cases of archival paraffin-embedded tumor specimens of newly diagnosed renal cell carcinoma (RCC) was studied retrospectively using rapid flow cytofluorometric (propidium iodide) DNA analysis. The degree of ploidy (DNA index), percentage of cells in the S-phase (SPF), and modal nuclear size were determined from histograms. The tumors were classified as diploid (DNA index = 0.9-1.1) or aneuploid. Proliferative activities of the tumors were assessed from the proportion of S-phase cells. The aneuploid occurrence was 77.6% in our series. Univariate and multivariate logistic regression analyses of DNA ploidy and associated parameters showed that DNA ploidy was correlated with SPF of cells (P = 0.0061) as no correlation was seen between DNA ploidy, sex, age, histological type, tumor size, stage, and nuclear size (P = 0.0697). Multiple aneuploid stem lines had no influences on ploidy and prognosis. Comparison of survival data using the multivariate stepwise hazard rate and Lee-Desu statistics showed that patient prognosis was closely related to tumor size (P = 0.006) and staging (P < 0.0001). DNA ploidy had marginal correlation to progression and disease-specific death (P = 0.064). Nevertheless, flow cytometric analysis in conjunction with conventional histopathology may have a potential role for the management of patients with RCC. PMID- 8231167 TI - IL-4 inhibits IL-2 induction of LAK cytotoxicity in lymphocytes from a variety of lymphoid tissues. AB - The purpose of this study was to determine the influence of recombinant human interleukin-4 (IL-4) on the regulation of lymphokine-activated killer (LAK) and mixed lymphocyte culture cytotoxic activity. Lymphocytes from several lymphoid tissues were studied including human peripheral blood leukocytes (PBL), spleen cells, thymocytes, and thoracic duct lymphocytes. Cells were cultured with IL-4 in the presence or absence of recombinant human interleukin-2 (IL-2) in 4-day cultures. LAK and natural killer (NK) activities were measured in a standard chromium release cytotoxicity assay against LAK-sensitive, NK-resistant M14 melanoma targets and NK-sensitive K562 erythroleukemic cells. IL-4 alone does not increase NK or LAK activity under the conditions studied. However, IL-4 does inhibit the induction of cytotoxic activity by IL-2. IL-4 inhibits IL-2-induced thymidine incorporation in 3-day PBL cultures, suggesting that the inhibition of cytotoxicity is not a dilution effect due to the proliferation of noncytotoxic cell populations. IL-4 also inhibits the development of LAK and NK-like activities generated in mixed lymphocyte culture (MLC) while augmenting MLC generated allospecific cytotoxic T lymphocyte activity. Thus, IL-4 appears to inhibit the induction of nonspecific cytotoxic effectors while augmenting the generation of MHC-specific responses. This confirms an important regulatory function for this lymphokine in the generation of cytotoxic effectors. PMID- 8231168 TI - Reducing the recurrence rate of duodenal ulceration after highly selective vagotomy: a study in the rat and in man. AB - Acid secretion after highly selective vagotomy (HSV) gradually increases with time and may be related to the rate of duodenal ulcer recurrence following this operation. Studies were carried out in the rat and in man to examine whether preventing recommunication of the vagal branches with their distal stumps reduces the rate of acid secretion recovery and duodenal ulcer recurrence after HSV. Male rats were allocated at random to three groups: (i) sham HSV, (ii) HSV, (iii) HSV with a polytetrafluoroethylene patch stitched against the denervated area of lesser curve. After housing for 2 years, the basal acid output of the HSV group was significantly (P < 0.01) depressed relative to control values (6.9 +/- 0.2 vs 14.1 +/- 0.4 mumole/hr, mean +/- SEM, n = 39 and 40, respectively). HSV with a patch had a basal acid output significantly (P < 0.01) lower than that of the HSV group (1.4 +/- 0.1 mumole/hr, mean +/- SEM, n = 41). Insulin significantly (P < 0.001) stimulated acid secretion in the HSV group but failed to do so in the group provided with a patch (1.4 +/- 0.1 vs 2.3 +/- 0.2 mumole/hr, mean +/- SEM, n = 41). Subcutaneous infusion of pentagastrin (4 micrograms/kg min) and carbachol (0.8 micrograms/kg min) for 24 hr produced duodenal ulceration in all of the control animals and in 20% of those in the HSV group, but in none of the HSV with a patch group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231169 TI - Effect of prolonged postischemic perfusion on neonatal hearts. AB - Recent research on myocardial preservation has emphasized the importance of events occurring early in the reperfusion period, which may be of less importance to myocardial outcome in the neonate. We therefore wished to study the contribution of prolonged postischemic perfusion. This situation may occur during repair of congenital heart defects when in an attempt to reduce the period of aortic cross-clamp, the surgeon completes right-sided repairs on a beating empty heart. We used an isolated working rabbit heart model to compare recovery after various periods of ischemia and postischemic perfusion. After 30 min in the working heart mode, hemodynamic measurements were made. The hearts were then rendered globally ischemic, with the exception of cardioplegia. At the conclusion of the ischemic period, the hearts were returned to the 37 degrees C chamber, and reperfused with buffer. After 30 or 60 min retrograde perfusion, the hearts were converted to the working mode again. The working heart period continued until the heart was no longer able to overcome its afterload ("pump failure"). We recorded the time to this point as survival time. Hearts were divided into four groups: Group I (n = 12), 60 min ischemia, 30 min nonworking perfusion; Group II (n = 11), 90 min ischemia, 30 min nonworking perfusion; Group III (n = 8), 60 min ischemia, 60 min nonworking perfusion; and Group IV (n = 11), 0 min ischemia, 130 min nonworking perfusion. Multivariate analysis showed that four factors influenced survival time: aortic flow, age, left ventricular end-diastolic pressure, and mean aortic pressure. Group II did not vary significantly from Group I.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231170 TI - Platelet-activating factor antagonism attenuates platelet and neutrophil activation and reduces myocardial injury during coronary reperfusion. AB - Platelet-activating factor (PAF) is known to be synthesized during tissue reperfusion and to be involved in the activation of platelets and neutrophils in inflammatory processes. The hypothesis of the present study is that PAF is central in the pathophysiology of myocardial reperfusion and that specific PAF receptor antagonism may reduce myocardial reperfusion injury. Utilizing an intact sheep model that involved a 90-min occlusion of the mid-left anterior descending coronary artery followed by 6 hr of reperfusion, a study group that received a specific PAF receptor antagonist (L-659,989, 5 mg/kg) 10 min before reperfusion was compared to a control group that received a saline placebo (n = 8 in each group). Coronary sinus platelet aggregating activity and neutrophil oxidative burst were studied by standard platelet aggregometry and the 2',7' dichlorofluorescein flow cytometric assay, respectively. Left coronary flow and left ventricular functions measured as peak +/- dp/dt and stroke work were analyzed. The extent of myocardial infarction at the end of 6 hr of reperfusion was measured by standard histochemical stainings. The results demonstrated that platelets were hyperaggregable and that neutrophil oxidative burst was increased in the myocardial compartment during the first 3 hr of coronary reperfusion after 90 min of ischemia. The administration of the PAF antagonist immediately before reflow effectively prevented the activation of platelets and neutrophils. This was associated with significantly improved coronary reflow and ventricular function during the observed reperfusion period and with reduced myocardial infarct measured at 6 hr of reperfusion. We conclude that the use of a specific PAF receptor antagonist, L-659,989, immediately before controlled coronary reflow attenuated the activation of platelets and neutrophils that occurred during reperfusion. These anti-platelet and anti-neutrophil effects together with the inhibition of the known direct deleterious effects of PAF on the myocardium translated into improved ventricular function and reduced myocardial infarct. PMID- 8231171 TI - Characterization of cardiac beta-adrenergic receptors in the guinea pig heart: application to study of beta-adrenergic receptors in shock models. AB - The myocardial response to catecholamines is significantly diminished in many types of shock or heart failure. The guinea pig heart is an ideal model for the study of shock, as it is relatively inexpensive, and the cardiovascular system of the guinea pig most closely resembles that of the human. Using this model, we have developed techniques to characterize and quantitate changes in beta adrenergic receptors (beta AR) in the guinea pig heart after burn injury. Preliminary experiments were performed to determine the optimum binding conditions, e.g., incubation time and conditions, protein concentrations, rinsing, etc. Additional experiments were conducted using agonists and antagonists to characterize the rank order of potency and stereospecificity of the beta AR. Crude membrane preparations (50 micrograms/250 microliters) from sham-burned and burned hearts were incubated with 8-10 concentrations of 125I cyanopindolol (10-450 pM) at 37 degrees C for 1 hr. Under these conditions, binding assays were linear with respect to protein concentration and time. Alprenolol (10 microM) was used to determine nonspecific binding. The membrane preparations used in this study bound both agonists and antagonists with a rank order of potency and stereospecificity characteristic of a beta-adrenergic receptor. Finally, agonist competition curves were performed with isoproterenol in the presence and absence of Gpp(NH)p to determine receptor regulation by the Gs protein. Analysis using computer-assisted techniques suggests that the fraction of high-affinity beta-receptors is significantly reduced after burn injury (41.2 +/- 4.7%) compared to sham-burned controls (54 +/- 2%, P < or = 0.023).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231172 TI - Failure of deferoxamine to reduce myocardial infarct size in a primate model of ischemia-reperfusion injury. AB - Baboons were subjected to treatment with deferoxamine (DF), a strong iron chelating agent, to inhibit the iron-dependent production of hydroxyl radicals. Studies were then done to determine if this would result in a reduction in the size of myocardial infarct. Baboons underwent occlusion of the left anterior descending coronary artery for 2 hr followed by reperfusion for the next 22 hr. A treated group (n = 4) received a 2-hr preischemic intravenous infusion of DF (10 mg/kg/hr). This infusion continued throughout the ischemic phase and 2 hr into the reperfusion phase. A control group (n = 8) underwent the identical protocol minus the DF infusion. At the end of the reperfusion period, the hearts were sectioned and stained for histological examination. The treated animals had a 22% larger volume of infarct compared with those of the controls (P = 0.06). There was no statistically significant difference (P > 0.05) in hemodynamic or epicardial ST segment measurements between the two groups. In this primate model, there was no myocardial protection afforded by DF. Baboons are similar to humans in that both have minimal collateral circulation. In the literature, DF has been noted to actually contribute to the production of free radicals in certain circumstances. This experiment appears to indicate that caution should be exercised in the use of DF in the treatment of ischemia-reperfusion injury of the heart. PMID- 8231173 TI - In vivo testing of an infection-resistant vascular graft material. AB - Present prosthetic arterial conduits continue to suffer the clinically and economically catastrophic complication of infection. We recently described a novel technique for binding quinolone antibiotics to Dacron based on principles of textile chemistry. This thermofixation procedure ("pad/heat") utilizes the limited fibrophilic characteristics of the quinolone antibiotic ciprofloxacin (Cipro) to permit pad/heat application and allowed controlled, sustained release from Dacron in several in vitro assays. The objective of this study was to test this infection-resistant prosthetic vascular graft material in an in vivo model. Dacron segments (1 cm2, either plain, dipped into antibiotic immediately prior to implantation, or Cipro pad/heat treated) were implanted in the dorsal subcutaneous tissue of the rabbit and directly contaminated with 10(6) Staphylococcus aureus. After 1 week, the samples were sterily harvested. Wounds were blindly graded on a scale from 1 (no evidence of infection, good tissue incorporation) to 4 (suppurative infection extending outside of the graft pocket, no gross tissue incorporation). Plain Dacron was easily infected in this model (mean grade 3.1 +/- 0.6, 92% culture positive). Notably, however, a significant (P < 0.05) wound grade difference between the dipped (2.3 +/- 1.0) and pad/heat (1.4 +/- 0.6) samples was demonstrated. Determination of adherent bacteria present on the implanted Dacron pieces by sonication and culture studies again revealed a significant difference between the dipped (56% culture positive) and pad/heat (12% culture) groups (P < 0.025). Histologic studies confirmed good tissue incorporation of the pad/heat samples. This project opens new avenues in the development of infection-resistant biomaterials. PMID- 8231174 TI - Mechanism of cellular phospholipid efflux. AB - Plasma phospholipid binding to cell-derived cholesterol is important in reverse cholesterol transport, a key step in the regression of atherosclerosis. However, the mechanism by which phospholipids are transferred from cells to plasma remains unclear. [3H]Choline-labeled phospholipid efflux from fibroblasts has been studied using plasma and its components as acceptors. The kinetics were resolved into a fast component (k1 = 0.119 +/- 0.23 min-1) that corresponded to high affinity binding of high-density lipoproteins (HDL) to the cell surface and a slow component (k2 = 0.0047 +/- 0.0009 min-1) due to protein-mediated desorption (n = 3). Altering the donor charge with heparinase or the acceptor charge by acetylation abolished the fast component, while the slow phase was unchanged. Only HDL displayed biexponential kinetics, comparable to whole plasma. Half-lives for low-density lipoprotein and very-low-density lipoprotein were t1/2 = 278 +/- 22 min and t1/2 = 1003 +/- 147 min, respectively. In the absence of transfer factor, HDL alone significantly reduced phospholipid efflux (t1/2 = 663 min). Phospholipid transfer protein restored biexponential kinetics. We conclude that cell membranes are a potentially important source of plasma phospholipids and that protein-mediated transfer to HDL is the major route for cell-to-plasma transfer. This step represents a locus for anti-atherosclerotic intervention. PMID- 8231175 TI - Inhibition of intestinal lipid peroxidation does not minimize morphologic damage. AB - Free radical-initiated lipid peroxidation (LP) following intestinal ischemia/reperfusion (I/R) may disrupt mucosal integrity. It is unknown if inhibition of LP prevents this injury. We analyzed rat ileum, subjected to I/R, for evidence of LP inhibition and structural damage following treatment with the 21-aminosteroid, U74389F, a potent LP inhibitor. Four groups of Lewis rats were studied after superior mesenteric artery occlusion with ligation of collateral arcades: (i) no ischemia, (ii) 10 min ischemia, (iii) 10 min ischemia + 1 hr reperfusion, (iv) 10 min ischemia + 1 hr reperfusion + U74389F (6 mg/kg i.v. prior to clamp removal and reperfusion). Ileal mucosa was analyzed for: 9i0 superoxide dismutase (SOD; U/mg protein), a key antioxidant enzyme, (ii) myeloperoxidase (MPO; U/mg protein), an index of PMN stimulation, (iii) malondialdehyde (MDA; pmole/mg), an end product of LP, and (iv) routine histology. MDA rose from 2.09 +/- 0.44 (mean +/- SE) in Group 1 to 15.10 +/- 2.22 in Group 3 following I/R (P < 0.01). In Group 2 and Group 4, MDA remained unchanged at 3.25 +/- 1.38 and 1.73 +/- 0.15, respectively. MPO, likewise, rose during I/R from 0.59 +/- 0.17 in Group 1 to 1.10 +/- 0.13 in Group 3 (P = 0.08) and 1.49 +/- 0.24 in Group 4 (P < 0.05). SOD did not vary significantly in the four groups studied. Despite PMN stimulation indicated by increased MPO with reperfusion, no PMN infiltration was seen histologically. U74389F normalized MDA, indicating effective inhibition of LP; however, similar epithelial sloughing and edema and hemorrhage in the lamina propria were seen in treated and untreated rats. These data implicate MDA-independent or possibly LP-independent pathways in intestinal morphologic damage occurring with I/R. PMID- 8231176 TI - Pharmacokinetics of DepoFoam gentamicin delivery system and effect on soft tissue infection. AB - Infections of burn and soft tissue wounds are often difficult to treat with systemic antibiotics since drug delivery to the wound may be suboptimal and high doses may result in toxicity. DepoFoam particles, a novel lipid-based drug delivery system, are composed of phospholipid membranes, enclosing multiple aqueous chambers into which pharmacologic agents can be encapsulated for local drug delivery. We encapsulated gentamicin (GENT) in DepoFoam particles with an average yield of 81% +/- 8 SD for 10 preparations. Encapsulated GENT was incubated in human plasma with t1/2 of 21 days, demonstrating stability in vitro. In vivo pharmacokinetics were determined by injecting CF-1 mice subcutaneously (sc) with a single dose of 0.5 mg of free (nonencapsulated drug) or DepoFoam GENT. At intervals postinjection the sc tissue was excised and blood was obtained by inferior cava puncture and both were assayed for GENT levels. At 0.5, 2, 6, and 24 hr following drug administration there was a significant difference between GENT levels in the tissue achieved with the encapsulated drug and free drug with n = 3-4 at each time point for each group (P < 0.01). By 24 hr following administration of free drug there was minimal detectable GENT in the tissues, while therapeutic levels of GENT remained in tissue at 24 hr following DepoFoam GENT injection. Serum GENT peaked at 30 min for both the DepoFoam (5 micrograms/ml) and free drug (10 micrograms/ml) and was undetectable by 2 hr (n = 3 each group).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231177 TI - Tolbutamide inhibits gluconeogenesis in the tumor-influenced hepatocyte. AB - The tumor-bearing state is associated with an increase in gluconeogenesis which may contribute to the development of cancer cachexia. The purpose of this study was to determine if tolbutamide, a drug known to decrease gluconeogenesis in diabetes, could decrease gluconeogenesis in hepatocytes isolated from tumor bearing rats. Hepatocytes from 24-hr fasted normal and methylcholanthrene-induced sarcoma-bearing rats (5-10% tumor burden) were isolated by in situ collagenase liver perfusion. Hepatocytes (n = 12 samples) from non-tumor-bearing (NTB) controls and tumor-bearing (TB) rats were incubated with lactate (10 mM) and alanine (10 mM) with and without 1 mM tolbutamide. Supernatant glucose concentration was measured at 30-min intervals for 2 hr. Rates of gluconeogenesis (+/- standard error) were calculated by linear regression and are expressed as nmole glucose/10(6) cells/min. Comparisons were made by two-way analysis of variance and significance defined as P < 0.05. TB hepatocytes had an increased rate of gluconeogenesis (P < 0.0001) from alanine and lactate (3.8 +/- 0.30 and 2.2 +/- 0.10, respectively) compared with NTB hepatocytes (0.66 +/- 0.10 and 1.2 +/- 0.04, respectively). TB hepatocytes treated with tolbutamide had a decreased (P < 0.0001) rate of gluconeogenesis from alanine and lactate (3.1 +/- 0.10 and 1.1 +/- 0.10, respectively) compared with untreated TB hepatocytes (5.3 +/- 0.10 and 2.1 +/- 0.10, respectively). Tolbutamide inhibits gluconeogenesis from lactate and alanine in tumor-influenced hepatocytes. PMID- 8231178 TI - Treatment of benign splenic cyst with splenic conservation. PMID- 8231179 TI - Ethical issues in the care of the elderly. PMID- 8231180 TI - Renal cell carcinoma detected as a result of blunt abdominal trauma. PMID- 8231181 TI - Acute hepatic decompensation in an alcoholic. PMID- 8231183 TI - Hospital charges in Tennessee. PMID- 8231182 TI - Frozen section error?--rare but disastrous. PMID- 8231184 TI - A Tennessee physician's view on health care reform. PMID- 8231186 TI - Prescription for workplace safety: bend and lift correctly to avoid back injuries! PMID- 8231185 TI - Managed care and the infamous hold harmless clause. AB - Hold harmless clauses may form the backbone of state and national health care reform since health insurers can effectively use them to pass off financial and liability risks to physicians. This is especially true in the TennCare proposal. Such clauses generally come in two forms, while a third type is often confused as a hold harmless clause, but is actually an indemnity provision. The first type involves a physician's agreement not to pursue a patient for fees in excess of those allowed in an HMO contract. The second type usually states that a physician will not look to an entity for a "contribution" to help pay off a claim for damages. The third type is not a hold harmless at all; rather, it requires a physician to "indemnify" an entity by paying its legal fees and any damages it is held liable for in a court judgment. Each clause holds increasingly dire consequences for physicians, and this should give them pause before they sign on the dotted line. Indemnity agreements are often excepted from professional liability coverage; therefore, a physician should consult with his carrier before agreeing to indemnify anyone. Finally, if such clauses must be included, a physician should demand extra consideration such as prompt payment and increased patient volume guarantees. This article examines some of the common types of hold harmless clauses the insurance industry expects physicians to sign under TennCare, and offers some negotiating points for physicians. PMID- 8231187 TI - Cardiac tumor presenting itself as pericardial tamponade in a child. PMID- 8231188 TI - Fournier's gangrene. PMID- 8231189 TI - In-office surgery?--same standards prevail! PMID- 8231190 TI - Injuries to the ascending aorta. PMID- 8231191 TI - Sudden infant death syndrome. PMID- 8231192 TI - Weakness in a young man. PMID- 8231193 TI - Crippling influenza. PMID- 8231194 TI - TMA leaders set the agenda for the future. TMA Communications and Public Service Committee. PMID- 8231195 TI - Home safety ... to go. PMID- 8231196 TI - The darker side of networking--price-fixing. PMID- 8231197 TI - TMA presidential address. PMID- 8231198 TI - Giants. PMID- 8231199 TI - Preoperative prediction for the use of cardiopulmonary bypass in lung transplantation. AB - Cardiopulmonary bypass has been widely used in the management of isolated single and double lung transplantations. Although there are certain clear-cut preoperative indications for cardiopulmonary bypass, in many patients the decision to use this modality is based on the hemodynamic consequences of intraoperative pulmonary artery clamping. We have performed 109 isolated lung transplantations. In 69 patients (38 single lung transplantations and 31 double lung transplantations) cardiopulmonary bypass was initiated only on the basis of intraoperative hemodynamics. We have analyzed preoperative data from these 69 patients to determine whether an intraoperative requirement for cardiopulmonary bypass can be predicted. Of 38 single lung transplantations, 12 necessitated cardiopulmonary bypass (all patients had restrictive lung disease). No patients with obstructive lung disease who underwent single lung transplantation required cardiopulmonary bypass (p < 0.001). For single lung transplantations, 6-minute walk, the arterial desaturation/oxygen requirements on exercise, and the right ventricular ejection fraction were all significantly different between the cardiopulmonary bypass and noncardiopulmonary bypass groups (p < 0.001). Of 31 double lung transplantations, 10 patients required cardiopulmonary bypass (seven had bronchiectasis, two had obstructive lung disease, and one had restrictive lung disease). For obstructive lung disease, no preoperative parameters predicted cardiopulmonary bypass. In conclusion, cardiopulmonary bypass is not necessary for most patients undergoing lung transplantation (in the absence of an absolute preoperative indication). Obstructive lung disease rarely necessitates cardiopulmonary bypass. In single lung transplantations, the subsequent requirement for cardiopulmonary bypass can be predicted from preoperative cardiopulmonary performance. For double lung transplantations, the requirement for cardiopulmonary bypass is usually dependent on unpredictable intraoperative factors. PMID- 8231200 TI - Bronchial contractions in transplanted lungs. Influence of denervation, acute rejection, and the bronchial epithelium. AB - The effects of pulmonary denervation and rejection on contractions of bronchial smooth muscle and epithelial modulation of these contractions were studied in dogs after denervation in right lung autotransplantation (n = 6) and acute rejection after right lung allotransplantation (n = 8). Immunosuppression was withdrawn from the latter group after 5 days; rejection developed after 3 additional days. A significant (p < 0.05) increase in mean peak airway pressure occurred with rejection of allotransplanted lungs. Rings cut from third-order bronchi of transplanted and contralateral unoperated (native) lungs in each animal were suspended in organ chambers for the measurement of isometric force. In some rings, the epithelium was removed mechanically. Acetylcholine (cholinergic neurotransmitter), serotonin (platelet-product), histamine (mast cell product), and endothelin-1 (endothelium-derived contracting factor) caused concentration-dependent contractions in all rings. In bronchi from native lungs, rings with epithelium contracted less than those without epithelium. This difference was lost after autotransplantation. The smooth muscle and epithelium were affected differently by autotransplantation. Contractions of rings without epithelium decreased in response to acetylcholine and endothelin-1, whereas contractions of rings with epithelium increased in response to histamine and 5 hydroxytryptamine (p < 0.05). During acute rejection, contractions were the same as those after autotransplantation. Bronchial content of endothelin increased fourfold with rejection. Relaxations to isoproterenol and prostaglandin E2 were similar in both groups. In conclusion, denervation reduced the ability of the smooth muscle to contract. The degree of acute pulmonary rejection seen in this study did not further affect bronchial contractions. Modulation of contractions by the bronchial epithelium was lost with both denervation and rejection. PMID- 8231201 TI - Temporary leukocyte depletion reduces ventricular dysfunction during prolonged postischemic reperfusion. AB - Leukocyte depletion improves early postischemic ventricular performance in neonatal models of global myocardial ischemia. However, the rate of leukocyte reaccumulation after cardiopulmonary bypass and its subsequent impact on myocardial function is not known. This laboratory study examined the effect of leukocyte depletion on myocardial performance during the initial 6-hour period after bypass in an in situ, in vivo porcine model of neonatal cardiac surgery. Fifteen 3- to 5-day-old piglets (eight control and seven leukocyte depleted animals) were instrumented by placement of left ventricular short-axis sonomicrometry crystals and an intraventricular micromanometer catheter. Mechanical leukocyte depletion was achieved with Pall RC100 filters (Pall Biomedical, Inc., Fajardo, Puerto Rico) in the cardiopulmonary bypass circuit. Neonatal hearts were subjected to 90 minutes of hypothermic ischemia after a single dose of cold crystalloid cardioplegia. Two control animals died after the operation and were excluded from data analysis. Leukocyte filtration reduced the granulocyte count during initial myocardial reperfusion to 0.8% of control values. However, circulating granulocyte counts increased in leukocyte depleted animals throughout the postoperative period, reaching 68% of control values by 6 hours. Despite this rapid return of circulating granulocytes, animals subjected to leukocyte depletion had significantly better preservation of left ventricular performance (measured by preload recruitable stroke work, p < or = 0.02), left ventricular systolic function (measured by end-systolic pressure-volume relationship, p < or = 0.05), and ventricular compliance (p < or = 0.04) during the experiment. These changes in ventricular function were associated with a significant increase in left ventricular water content (p < or = 0.02) and tissue myeloperoxidase activity (p < or = 0.005) in control animals compared with leukocyte depleted animals. This study demonstrates that leukocyte depletion during initial reperfusion results in sustained improvement in postischemic left ventricular function despite the rapid return of granulocytes to the circulation. PMID- 8231202 TI - Long-term heart preservation by intermittent perfusion with crystalloid medium. AB - This study was undertaken to determine whether hearts preserved with intermittent coronary perfusion would recover physiologic function after a prolonged period of hypothermic preservation. Intermittent perfusion is commonly used for cardioplegia, but its efficacy in long-term heart preservation has not yet been demonstrated. Five groups of isolated rat hearts were studied (n = 7 per group): (1) fresh nonpreserved control hearts; (2) hearts preserved with continuous low pressure perfusion via the aorta; (3) hearts preserved with cycles of 5 minutes of perfusion followed by 25 minutes of nonperfusion; (4) hearts preserved with cycles of 10 minutes of perfusion followed by 25 minutes of nonperfusion; (5) hearts preserved with submersion storage without perfusion. An oxygenated extracellular-type crystalloid medium (oxygen tension = 820 +/- 5 mm Hg) was used as a preservation medium; preservation was for 12 hours. During preservation, the coronary resistance of the intermittent perfusion-preserved hearts increased significantly, and these hearts produced significantly more excess lactate than did hearts in the other two preservation groups. The submersion-stored hearts exhibited no postpreservation ventricular function in an isolated perfused working rat heart system. The poststorage function of the other four groups, which was quantified during a 4-hour, 37 degrees C perfusion period at constant heart rate, indicated that there were no significant group differences with respect to output or energetics (coronary flow, aortic output, cardiac output, myocardial oxygen consumption, and external work efficiency). The intermittent perfusion-preserved hearts had significantly lower postpreservation contractile function (left ventricular systolic pressure, peak rates of left ventricular pressure development and relaxation, peak aortic flow rate, stroke work, and peak power) and higher left ventricular end-diastolic pressure compared with the control group. Although hearts preserved with intermittent perfusion had a loss of contractile function and decreased compliance compared with fresh hearts, after preservation they had better function than did hearts preserved with submersion storage and the same function as hearts preserved with continuous perfusion. PMID- 8231203 TI - Cerebral vascular reactivity to carbon dioxide before and after cardiopulmonary bypass in children with congenital heart disease. AB - We examined cerebral vascular reactivity to carbon dioxide before and after cardiopulmonary bypass in 15 pediatric patients aged 2 to 9 years undergoing cardiac operations. Cerebral vascular reactivity was noninvasively assessed by transcranial Doppler ultrasonography. The cerebral blood flow velocity was plotted as a function of arterial carbon dioxide partial pressure. An exponential function was fitted for these plots and an exponent of curve was defined as the cerebral vascular reactivity to carbon dioxide. There was no significant change in this reactivity after cardiopulmonary bypass as compared with before bypass (before 0.028 +/- 0.003; after 0.030 +/- 0.016). For the entire series, we obtained best-fit curves of y = 2.8e0.046x (r = 0.91) and y = 3.4e0.031x (r = 0.89) (x; arterial carbon dioxide partial pressure, y; percent changes of cerebral blood flow velocity) before and after cardiopulmonary bypass. We conclude that the cerebral vascular reactivity to carbon dioxide was preserved before and after cardiopulmonary bypass in children undergoing cardiac operations. These results suggest that cerebral perfusion before and after cardiopulmonary bypass is not critically compromised. PMID- 8231204 TI - Activation of fibrinolysis in the pericardial cavity during cardiopulmonary bypass. AB - The clotting and fibrinolytic systems are activated by tissue factor and by tissue-type plasminogen activator in the pericardial cavity, where the thrombogenicity is greater than that of the surface of modern extracorporeal circuits. This local activation may have consequences for the systemic activation processes during cardiopulmonary bypass. To test this hypothesis, we investigated blood activation by interrupting the blood suction from the pericardial cavity during cardiopulmonary bypass in clinical coronary artery bypass operations. In blood collected in the pericardial cavity, thrombin-antithrombin III complex (p < 0.01), tissue-type plasminogen activator antigen (p < 0.05), fibrinogen degradation products (p < 0.01), and fibrin degradation products (p < 0.01) were significantly higher than in the systemic blood. Plasma heparin was significantly consumed in the pericardial cavity (p < 0.01). Once the pericardial blood was returned to the systemic circulation after resumed suction during cardiopulmonary bypass, thrombin-antithrombin III complex (p < 0.05), fibrinogen degradation products (p < 0.05), and fibrin degradation product (p < 0.05) concentrations increased significantly in the systemic blood. The effects of pericardial tissue on activation of clotting and fibrinolysis were also studied in vitro. When human plasma was incubated for 5 minutes with rabbit pericardium at reduced heparin concentrations, we found significant generation of thrombin (p < 0.05) and plasmin (p < 0.05). If the thrombin inhibitor hirudin was added, plasmin generation was also inhibited (p < 0.05). The results of the clinical and experimental study are in agreement with our hypothesis that tissue factor and tissue-type plasminogen activator accelerate the activation of clotting and sequentially of fibrinolysis under conditions of low heparin concentrations in the pericardial cavity and that this local activation contributes highly to the systemic activation, affecting hemostasis during cardiopulmonary bypass. Topical use of heparin in the pericardial cavity therefore seems indicated to reduce blood activation during cardiopulmonary bypass. PMID- 8231205 TI - Ro 44-9883, a new nonpeptide glycoprotein IIb/IIIa antagonist, prevents platelet loss during experimental cardiopulmonary bypass. AB - Extensive contact between blood and artificial surfaces causes platelet activation and depletion. The aim of the study was to test the efficacy of Ro 44 9883, a new nonpeptide, reversible, glycoprotein IIb/IIIa antagonist in preventing platelet count drop in dogs undergoing cardiopulmonary bypass during 2 hours. Twenty-two heparinized dogs were divided into three groups, one control group (n = 9) not treated, one group treated with a low dose of Ro 44-9883 (145 micrograms/kg intravenously, n = 6), and one group treated with a high dose of Ro 44-9883 (870 micrograms/kg intravenously, n = 7). In the control group, platelet counts declined to 53% +/- 15% of initial levels at the start of cardiopulmonary bypass and remained lower than 80% of initial levels during follow-up. Platelet count drop was completely prevented in the Ro 44-9883 high-dose group, whereas it was only partially prevented in the low-dose group. Blood loss was similar in the three groups despite the fact that bleeding times were longer in the Ro 44-9883 high-dose group than in the control group. We conclude that Ro 44-9883, together with heparin as a standard anticoagulant, is highly effective in preventing platelet count drop during cardiopulmonary bypass without causing excessive bleeding. PMID- 8231206 TI - New configuration of double cardiomyoplasty based on studies of the length tension properties of the latissimus dorsi muscle. AB - A new configuration of double cardiomyoplasty was designed according to studies of the length-tension properties of the linear latissimus dorsi muscle. Four dogs had both their right and left latissimus dorsi muscles dissected from the chest wall and attached to a tensiometer to measure force of contraction. The maximum active tension obtained with stimulation of the linear latissimus dorsi muscle was observed when the muscle was at its resting anatomic length and up to 5% above this length. Eight dogs had a double cardiomyoplasty in which the resting anatomic length of both muscles was maintained. Control hemodynamic parameters obtained with the muscles at rest were compared with stimulated muscle protocols. In a normal heart state, stimulation of the double cardiomyoplasty increased the cardiac output 32% (p < 0.05), the stroke volume 39% (p < 0.05), and the left ventricular pressure 42% (p < 0.05). When acute heart failure was induced with high-dose intravenous propranolol (5 mg/kg), stimulation of the double cardiomyoplasty increased the cardiac output 32% (p = 0.01), the stroke volume 32% (p < 0.05), rate of pressure rise 39% (p < 0.01), and myocardial thickening 39% (p < 0.01). The study demonstrated that this configuration of double cardiomyoplasty provides significant hemodynamic assistance in the normal and acutely failing canine heart. PMID- 8231207 TI - Superiority of extended en bloc esophagogastrectomy for carcinoma of the lower esophagus and cardia. AB - The belief that transhiatal esophagogastrectomy results in the same survival as a more extensive en bloc resection was tested in 69 patients with carcinoma in the distal esophagus and gastric cardia. Preoperative and intraoperative staging defined three distinct subgroups of patients. Those with apparently limited disease and good general health (group I, n = 30) underwent en bloc resection. Those with apparently limited disease but poor physiologic reserve (group II, n = 16) underwent transhiatal resection, as did those with evidence of more advanced disease (group III, n = 23). Overall, survival was significantly better in the 30 patients who underwent en bloc resection (41%) than in the 39 patients who underwent transhiatal resections (14%; p < 0.001, log-rank). Clinical staging showed apparently limited disease in 46 patients (groups I and II). These groups differed only in the presence of poor physiologic reserve because the percentages of patients with tumors limited to the esophageal wall (group I 13/30, group II 6/16) and four or fewer lymph node metastases (group I 21/30, group II 15/16) at the time of pathologic staging were not significantly different. Survival after en bloc resection was, however, significantly better (41% versus 21%; p < 0.05, log-rank). According to the WNM system of pathologic staging, 19 patients had early lesions defined as intramural lesions associated with four or fewer lymph node metastases, 26 had intermediate lesions defined as either transmural or associated with more than four lymph node metastases, and 24 had late lesions defined as both transmural and associated with fewer than four lymph node metastases. Survival was significantly better in patients with early lesions after en bloc resection compared with transhiatal resection (75% versus 20%, p < 0.01), survival was also significantly better in patients with advanced lesions (27% versus 9%, p < 0.01). For intermediate lesions, the survival was similar (14% versus 20%), although the median survival after en bloc resection was longer (24 months versus 8 months). PMID- 8231208 TI - Short-segment intestinal interposition of the distal esophagus. AB - Esophageal replacement remains a challenge. Colon and jejunum provide alternative conduits to replace the lower esophagus when stomach is not suitable. Between 1971 and 1991, 41 patients underwent short-segment interposition of the esophagus with jejunum or colon. Indications were failed antireflux procedures (n = 21), nondilatable stricture (n = 9), achalasia (n = 2), moniliasis (n = 2), Barrett's esophagus with carcinoma in situ (n = 2), hemorrhagic esophagitis after esophagogastrectomy (n = 1), motility disorder (n = 1), instrumental perforation (n = 1), carcinoma (n = 1), and leiomyosarcoma (n = 1). Thirty-one patients (75.6%) had prior surgical procedures. Interposition with colon was performed in 22 patients and with jejunum in 19. Major complications occurred in 45% after colon interposition (10/22) and hospital mortality was 4.5% (1/22). Major complications after jejunal interposition occurred in 31% (6/19) and hospital mortality was 10.5% (2/19). A contained anastomotic leak occurred in 1 patient, perforation of a colon segment in 1, and jejunal graft necrosis in a third. Late functional results in 34 patients with a mean follow-up of 87 months were excellent or good in 26, fair in 5, and poor in 1. Colon interposition failed to improve symptoms in 2 patients with gastrointestinal motility disorders. Six patients underwent manometry and barium food provocation study. Two colon segments and 3 jejunal interpositions were hypoperistaltic or aperistaltic according to manometry. There was 1 case of aperistaltic jejunum with a distended afferent loop. When stomach is not available, successful palliation of swallowing can be accomplished with either jejunum or colon. Surgeons involved in the management of esophageal disease should be familiar with the technical details of both procedures. PMID- 8231209 TI - Pulmonary resection after pneumonectomy in patients with bronchogenic carcinoma. AB - Eight patients with a previous pneumonectomy for bronchogenic carcinoma underwent an additional resection because of a second primary carcinoma in the remaining lung. One patient died of pulmonary embolism in the postoperative period. The postoperative course was otherwise uneventful except for prolonged air leak. Two patients died after 3 months (bone metastasis) and 5 months (recurrent small-cell carcinoma). Two patients were alive at the time this article was written but had evidence of recurrence after 18 months (distant metastasis) and 21 months (local recurrence at the site of positive resection margins). Three patients were alive and doing well without evidence of disease after 16, 17, and 40 months. After careful selection, even patients with a previous pneumonectomy may be good candidates for additional resection of a second primary bronchogenic carcinoma. PMID- 8231210 TI - Results of pulmonary resection of metastatic colorectal cancer and its application. AB - A resection of pulmonary metastases from colorectal cancer was performed in 27 patients. The cumulative 5-year survival after thoracotomy was 41.1%. Patients with either one or two metastases had a significantly better survival, with a 5 year survival of 54.3%, than did those with a greater degree of metastases (p < 0.01). Other factors including the disease-free interval, size of metastases, type of pulmonary resection, and stage of primary cancer had no apparent influence on post-thoracotomy survival. Furthermore, the presence of controlled hepatic metastases had no adverse effect on survival and did not seem to contraindicate pulmonary resection. The lung and primary colorectal region were the major sites of recurrence. Therefore, the primary colorectal site should be explored thoroughly and the number of metastases should be carefully considered when pulmonary resection is considered for metastatic colorectal cancer. PMID- 8231211 TI - Correction of total anomalous pulmonary venous connection in infancy. AB - From January 1985 through January 1993, 41 patients less than 1 year of age underwent operative correction of isolated total anomalous pulmonary venous connection. There were 24 boys and 17 girls. The median age at operation was 13 days (range 1 to 282 days) and weight was 3.6 kg (2.5 to 5.2 kg). Locations of the connections were supracardiac in 19, cardiac in 9, infracardiac in 11, and mixed supracardiac and cardiac in 2. Obstruction of the pulmonary veins was severe in 24, mild in 3, and absent in 14. Preoperative stabilization included mechanical ventilation for 15 patients for a mean duration of 2 1/2 days and extracorporeal membrane oxygenation for 1 patient for 1 day. All operations were performed with deep hypothermia and circulatory arrest (mean arrest time 34 minutes). Supracardiac connections were repaired by performing a side-to-side anastomosis between the pulmonary venous confluence and the dome of the left atrium through a superior approach between the superior vena cava and the aorta. Coronary sinus connections were repaired by enlarging the atrial septal defect and the coronary sinus communication with the left atrium and closing the atrial defect with a large patch. Infracardiac repairs included elevation and rotation of the heart to the right and an elongated side-to-side anastomosis between the common venous confluence and the left atrium. One patient died 1 week postoperatively of persistent pulmonary hypertension. Another patient, who was supported by extracorporeal membrane oxygenation before the operation, died 3 months after the operation as a consequence of pulmonary lymphangiectasia. All other patients are alive and well with a mean follow-up of 26 months (range 3 to 77 months). One patient required two subsequent reoperations for persistent pulmonary venous obstruction, and another patient had superior vena cava obstruction necessitating reoperation. Operative treatment of total anomalous pulmonary venous connection in infants can be performed with low mortality and an infrequent need for reoperations. PMID- 8231212 TI - Management of complications of Sideris transcatheter devices for atrial septal defect closure. AB - Various devices that can be inserted transvenously to close an ostium secundum atrial septal defect are undergoing clinical trials. Although these are safe and effective in most instances, they may occasionally dislodge or fail to "button" properly, causing migration and embolization. We report two cases in which the occluder and counteroccluder of the Sideris device for transvenous atrial septal defect occlusion (Custom Medical Devices, Amarillo, Tex.) failed to "button" appropriately, migrating in the right atrium in one patient and embolizing to the pulmonary artery in the second patient. An emergency operation was required to retrieve the device and repair the atrial septal defect. PMID- 8231213 TI - Coarctation of the pulmonary artery in neonates. Prevalence, diagnosis, and surgical treatment. AB - Stenosis or discontinuity of the central pulmonary artery at its bifurcation is often found in patients with congenital heart disease with intracardiac obstruction of the right side. The prevalence of this disease in autopsy cases is about 30%. The clinical prevalence has not been established yet because early diagnosis of this lesion is easily missed. Embryology suggests an etiologic similarity between coarctation of the aorta and this type of stenosis: both are caused by invasion of ductal tissue into the respective great artery. Therefore, the term coarctation of the pulmonary artery is appropriate. To recognize or exclude coarctation of the pulmonary artery, we examined left ventricular angiograms, aortograms, or both in the hepato-clavicular++ view in 25 consecutive neonates with severe arterial hypoxemia caused by right ventricular outflow tract obstruction. The prevalence of coarctation of the pulmonary artery was 36% (9/25). Fifteen neonates had two ventricles, and the other 10 had different types of univentricular heart. In eight cases no patent ductus arteriosus was found, and in none of these cases was coarctation of the pulmonary artery present. Conversely, in 9 of 17 neonates with a patent ductus arteriosus, coarctation of the pulmonary artery of different degree was demonstrated, even during prostaglandin E1 infusion in 7 neonates. With regard to surgical procedures, ductal tissue in the pulmonary artery wall should be completely removed. The aim is to establish adequate blood flow to both right and left pulmonary arteries. In this way unilateral hypoplasia of the arterial tree and consecutive growth of tortuous bronchial collaterals is prevented. PMID- 8231214 TI - An analysis of risk factors for death and mode-specific death after aortic valve replacement with allograft, xenograft, and mechanical valves. AB - From September 1967 to January 1990, a total of 2100 patients underwent 2366 aortic valve replacements with a variety of allograft, xenograft, and mechanical valves. Concomitant procedures were performed in 764 patients. Actuarial survival at 12 years was 59.6% (70% confidence limits 57.8% to 61.4%). Hazard function for death was highest immediately after operation, falling to merge with a slowly rising phase of risk at approximately 3 months. Actuarial freedom from sudden death at 12 years was 88.0% (70% confidence limits 86.7% to 89.3%). The shape of the hazard function for sudden death was similar to that for death. Actuarial freedom from death with cardiac failure at 12 years was 87.9% (70% confidence limits 86.5% to 89.2%). The shape of the hazard function for death with cardiac failure was also similar to that for death. Risk factor analysis revealed the important deleterious impact on long-term survival resulting from impaired left ventricular structure and function because of aortic valve disease. No current era valve used in this study (allograft, xenograft, or mechanical) was a risk factor for death. Both aortic wall disease and endocarditis necessitating aortic valve replacement substantially decreased long-term patient survival. Aortic valve replacement is advisable much earlier in the natural history of aortic valve disease before secondary left ventricular damage occurs. PMID- 8231215 TI - Effect of cryopreservation on the presence of endothelial cells on human valve allografts. AB - Cryopreserved human allograft valves are useful in a variety of cardiac operations. The presence or absence of endothelial cells on allografts may be important in determining immunogenicity and ultimate graft longevity. The purpose of this study was to determine whether endothelial cells are present on cryopreserved human allografts. Portions of cryopreserved allografts (35 valve leaflets, 96 pieces of arterial wall) not used at operation were studied. For comparison, untreated tissues (44 valve leaflets, 46 pieces of arterial wall) were obtained from structurally normal hearts and lungs removed or inserted at the time of transplantation and from pathologic tissues obtained during operations for congenital heart defects. A monolayer of cells from the luminal surface of each specimen was harvested by means of a Hautchen preparation. The monolayer was stained with fluorescein isothiocyanate-labeled Ulex europaeus I, a lectin with strong affinity for human endothelium. Positive staining with fluorescein was considered to be evidence for the presence of human endothelium. Endothelial cells were observed on 21 of 131 (16%) cryopreserved allograft specimens and on 70 of 90 (78%) untreated tissues (p < 0.001). These results show that cryopreservation typically results in the loss of endothelium from aortic and pulmonary valve allografts. These findings may have important implications for the immunologic response of the host to allograft implantation. PMID- 8231216 TI - The Bjork-Shiley Monostrut valve. Clinical experience in 647 patients. AB - A total of 647 consecutive patients underwent valve replacement with a Bjork Shiley Monostrut valve (Shiley, Inc., Irvine, Calif.) from January 1, 1984, through December 31, 1988. A total of 135 patients (20.8%) had had a previous cardiac correction. Overall hospital mortality was 6.8%. Preoperative functional class, cardiopulmonary bypass time, aortic crossclamping time, and cause of valve replacement influenced in-hospital mortality significantly (univariate analysis). The median follow-up was 30 months. Overall actuarial survival was 81% +/- 4% at 6 years. Univariate and multivariate regression analysis showed that preoperative functional class, total cardiopulmonary bypass time, size of mitral prosthesis, and pure mitral insufficiency significantly influenced total mortality after valve replacement. Additional coronary bypass grafting and redo valve replacement did not. The actuarial rate of freedom from thromboembolism was 86% +/- 4% at 6 years. There were no cases of valve thrombosis. Patients having a thromboembolic event showed a higher probability of late death after native valve replacement. PMID- 8231217 TI - Early experience with the inferior epigastric artery in coronary artery bypass grafting. A word of caution. AB - The excellent results with the internal thoracic artery for coronary artery bypass grafting have prompted the search for other arterial conduits. From November 1991 to February 1992, 18 patients underwent coronary artery bypass grafting with the use of inferior epigastric artery grafts. Patients' ages averaged 52 +/- 9 years. Bilateral internal thoracic artery grafts were used in 17 patients (17/18, 94%) and a free graft with one inferior epigastric artery was used in each patient. The inferior epigastric artery grafts were anastomosed to the right coronary artery (n = 9), a marginal circumflex artery (n = 4), and to a diagonal artery (n = 5). Three patients had abdominal wound complication related to harvesting of the inferior epigastric artery. Immediate postoperative angiographic evaluation of inferior epigastric artery grafts showed that eight grafts were patent (8/14, 57%). Four of the occluded inferior epigastric arteries were grafted to the right coronary artery and one to the second marginal circumflex coronary artery. Because of the low patency rate of inferior epigastric artery grafts, a word of caution is necessary in the selection of patients. At the present time, the inferior epigastric artery appears to be an interesting alternative only in patients who have no other available conduits. PMID- 8231218 TI - Fibrinogen receptor antagonists in cardiopulmonary bypass. PMID- 8231219 TI - Concurrent quadruple-valve percutaneous balloon valvuloplasty. PMID- 8231220 TI - Early but definitive correction of symptomatic coarctation of the aorta. PMID- 8231221 TI - Invited letter concerning: Dynamic cardiomyoplasty acutely impairs left ventricular diastolic function. PMID- 8231222 TI - Isolated coronary artery bypass grafting in one hundred octogenarian patients. PMID- 8231223 TI - Qualitative microscopy of implanted vein grafts. PMID- 8231224 TI - Surgical angioplasty of the left main coronary artery in a 12-year-old child. PMID- 8231225 TI - Bifurcated ("Y") internal thoracic-coronary artery grafts. PMID- 8231226 TI - Management of endobronchial hemorrhage and pulmonary artery pseudoaneurysm. PMID- 8231227 TI - Frequent detection of human herpesvirus-6 sequences by polymerase chain reaction in paraffin-embedded lymph nodes from patients with angioimmunoblastic lymphadenopathy and angioimmunoblastic lymphadenopathy-like lymphoma. AB - In search of a possible involvement of viral agents in angioimmunoblastic lymphadenopathy with dysproteinaemia (AILD) and AILD-like lymphoma (AILD-L), we studied the presence of human herpesvirus-6 (HHV-6) in the lymph node biopsies of 12 cases by polymerase chain reaction (PCR). Given the rarity of this lymphoproliferative disorder, we investigated archival specimens, consisting of formalin-fixed and paraffin-embedded tissues, obtained from patients with a clinical and histologic diagnosis of AILD and AILD-L. HHV-6 sequences were detected in the lymph node biopsies of 7 out of the 12 AILD and AILD-L cases examined. HHV-6 sequences were identified also in the involved liver and spleen tissues of one patient and in the PBMCs of two patients, all carrying viral sequences in the affected lymph nodes. We also used PCR to characterize the HHV-6 genomes, showing that two different viral strains are represented in the pathologic tissues. This study provides evidence of the presence of HHV-6 specific sequences in an unexpectedly high proportion of our series of AILD and AILD-L cases, suggesting a possible involvement of this lymphotropic virus in the pathogenesis of at least some cases of the disease. PMID- 8231228 TI - A new pharmacological function of phthalate ester plasticizer. PMID- 8231229 TI - Induction of differentiation of HL-60 and WEHI-3B D+ leukemia cells by lithium chloride. PMID- 8231230 TI - A role for mitochondrial DNA in the pathogenesis of radiation-induced myelodysplasia and secondary leukemia. AB - The onset of acute myeloid leukemia following ionizing radiation or alkylating agent exposure is antedated months to years by the development of 'preleukemia', or secondary myelodysplastic syndrome (sMDS). Mitochondrial abnormalities induced by chloramphenicol and clonal deletions of mitochondrial DNA (mt DNA) in the bone marrow create hematological defects similar to sMDS, and abnormal dimers of mt DNA are observed in acute leukemia. This suggests a role for mt DNA in the pathogenesis of sMDS and secondary leukemia. We outline disparate experimental evidence to support this concept and suggest a role for select protease inhibitors in the clinical management of this disorder. PMID- 8231231 TI - Hypoxaemia in patients with hyperleukocytosis: true or spurious, and clinical implications. AB - It has been suggested that in asymptomatic patients with leukaemias and very high white blood cell counts, the apparent hypoxaemia found using routine blood gas analysis is spurious, the result of excessive O2 metabolism by leukocytes. Pulse oximetry has been suggested as a means of overcoming the shortcomings of blood gas analysis in the assessment of these patients. We present the findings of two patients with extremely high white cell counts, which show that the hypoxaemia found is in fact true hypoxaemia, even in asymptomatic patients, and that met haemoglobinaemia may be at least in part responsible for the low PaO2. We also showed that pulse oximetry was completely unreliable in our patients due to the elevated met-haemoglobin levels. We recommend that all patients with markedly elevated white cell counts should undergo blood gas analysis with no delay between sampling and processing and that patients with low PaO2 should undergo urgent cytoreduction. PMID- 8231232 TI - De novo myelodysplastic syndrome (MDS) with deletion of the long arm of chromosome 20: a subtype of MDS with distinct hematological and prognostic features? AB - We report on 8 cases of de novo myelodysplastic syndromes (MDS) with deletion of the long arm of chromosome 20 (del 20q), who represented about 2% (8/392) of our cases of de novo MDS with cytogenetic analysis seen during a period of 9 yr. Median age was 69 yr, and there were 7 males and 1 female. Anemia was absent or very mild (Hb > 11 g/dl) in 5 patients. Only 1 patient had neutrophils < 0.5 x 10(9)/l, and none had platelets < 50 x 10(9)/l. Four patients had refractory anemia (RA), 2 had refractory anemia with ringed sideroblasts (RARS), and 2 had refractory anemia with excess of blasts (RAEB). Del 20q was isolated in 5 patients, and associated with other chromosomal rearrangement(s) in 3 patients. Only 1 patient progressed to ANLL and 2 showed an increase in bone marrow blasts during evolution. The 5 other patients had stable disease after 18-77 months. By comparison with de novo MDS patients with other cytogenetic findings, patients with del 20q had a tendency towards lower incidence of anemia and excess of marrow blasts, lower incidence of progression to AML and more prolonged survival, although differences were not significant. Only patients with isolated del 5q had a more prolonged survival than patients with del 20q. PMID- 8231233 TI - Variable growth in vitro of chromosomally different stem cells in myelodysplastic syndromes. AB - Ten patients with myelodysplastic syndromes were investigated using in vitro colony assay of bone marrow cells and chromosome analysis of single colonies. The result was compared with conventional cytogenetic analysis of bone marrow cells. The chromosome abnormalities included were 5q-, +8, -7, 11q-, -Y and one complex karyotype. Erythroid colony formation was reduced in eight patients, while the number of granulocyte-macrophage colony-forming units was normal or increased. Cytogenetic examination of single colonies showed that both chromosomally normal and abnormal stem cells had colony forming ability. The proportion of cytogenetically abnormal colonies varied between 25 and 100% of analysed colonies. No further clones than those found in direct cytogenetic analysis were revealed after in vitro growth. One patient with a 5q- abnormality and one patient with a -7 abnormality showed a significantly lower proportion of cytogenetically abnormal colonies than the proportion of abnormal cells in the direct bone marrow chromosome preparation. One patient with a +8 abnormality showed a growth advantage of the +8 clone in comparison with the chromosomally normal clones, but this was not statistically significant. In two patients a chromosomally changed stem cell gave rise to both erythroid and myeloid colonies. The FAB-class did not seem to influence the growth of either chromosomally normal or abnormal colonies. PMID- 8231234 TI - Higher proliferative response in B-chronic lymphocytic leukemia (B-CLL) as compared to B-monoclonal lymphocytosis of undetermined significance (B-MLUS) after stimulation with Staphylococcus aureus and anti-CD40 monoclonal antibodies. AB - B-CLL is a malignant monoclonal B-cell disorder and B-MLUS is the benign counterpart. The proliferative response and the capacity to secrete IgM was measured in B-CLL and B-MLUS, respectively, and compared to normal B-cells. SAC and a mAb against CD40 were used as stimulatory agents. No cell population responded to anti-CD40 mAb alone. SAC only induced a high DNA synthesis rate in normal B-cells as well as in B-CLL cells, although the magnitude was three-fold lower and delayed for about 48 h in B-CLL. B-MLUS cells did not proliferate in response to SAC. The combination of anti-CD40 and SAC enhanced the proliferative capacity of normal B-cells and produced a more rapid response in B-CLL. B-MLUS cells were not activated. Normal B-cells and B-MLUS did not secrete IgM after SAC stimulation, while B-CLL cells had a continuous increase in the IgM production during a 6-day culture period. The higher proliferative capacity of B-CLL cells compared with B-MLUS cells may be explained by an increased expression of activation molecules e.g. receptors for various cytokines and growth factors. Moreover, the inertness and inability of B-MLUS cells in comparison to normal B- and B-CLL cells to respond to powerful activation signals might indicate an intrinsic defect of B-MLUS cells in the signal transduction leading to a block of mitosis and a benign course of the disease. PMID- 8231235 TI - Expression and modulation of annexin VIII in human leukemia-lymphoma cell lines. AB - Annexin VIII is a calcium- and phospholipid-binding protein with anticoagulant activity. Annexin VIII mRNA was found to be specifically expressed in acute promyelocytic leukemia (APL) cells; it was not found in other types of acute myeloid leukemia (AML) nor in lymphoid malignancies. Using Northern blot analysis we investigated annexin VIII expression in 142 continuous human leukemia and lymphoma cell lines at the mRNA level. While the only APL cell line, NB-4, was indeed positive, other cell lines also displayed annexin VIII mRNA: 4/22 myeloid cell lines, 8/23 monocytic cell lines, 2/8 megakaryoblastic cell lines, 5/26 lymphoma-derived cell lines, 2/10 myeloma cell lines and 1/44 lymphoid leukemia cell lines. The strongest expression was seen in NB-4 and in the Hodgkin's disease derived cell line HDLM-2. Treatment of NB-4 cells with all-trans retinoic acid (ATRA) or the phorbol ester TPA induced terminal differentiation and down regulated annexin VIII mRNA expression rapidly within a few hours; vitamin D3 was ineffective in this regard; the protein kinase C activator Bryostatin 1 up regulated the expression. A panel of initially negative cell lines could not be induced by any of these biomodulators to transcribe annexin VIII. The half-life (T1/2) of annexin VIII mRNA was about 3-4 h using actinomycin D as transcription inhibitor. Treatment with ATRA or TPA prior to exposure to actinomycin shortened the T1/2 to 2 h while Bryostatin 1 extended it to 6h. As 21/141 non-APL cell lines were positive, annexin VIII cannot be used as a marker gene for APL cells; however, it might be associated with myelomonocytic or erythro-megakaryoblastic precursor cells. Annexin VIII gene expression might play a unique role in the proliferation and/or differentiation of leukemic cells and could be associated with the particular abnormal hemostasis of some leukemias. PMID- 8231236 TI - Multiple pre-neoplastic events and clonal selection of radiation induced mouse thymic lymphomas shown by TCR gene rearrangements. AB - After split-dose irradiation, pre-lymphoma cells develop from a tumor-specific surface antigen TL-2+ thymocyte subpopulation. To analyze the clonality of pre lymphoma cells, various numbers of TL-2+ thymocytes from a single irradiated mouse were intra-thymically injected to Thyl congenic recipient mice. The incidence of donor type thymic lymphoma(s) was subsequently examined in a group of recipient mice. We chose several lymphomas derived from a single donor mouse and analyzed the TCR gene rearrangements and V(D)J junctional diversity as genetic markers of clonality. These results indicate multiple initial neoplastic events and clonal selection into lymphoma. PMID- 8231237 TI - Cyclophosphamide treatment of an SJL murine B-cell lymphoma increases the proportion of suppressive CD8+ over tumor-stimulatory CD4+ T-lymphocytes. AB - Lymphomas (formerly reticulum cell sarcomas (RCS)) which develop spontaneously in SJL mice are a murine counterpart of human low grade B-cell non-Hodgkin's lymphoma. Tumor cells stimulate proliferation of syngeneic CD4+ T-lymphocytes which secrete cytokines required for tumor growth. Cyclophosphamide treatment of tumor-bearing mice (RCS/Cy) decreases in vitro tumor-stimulated CD4+ T-cell proliferation and, in turn, tumor growth, in part, through the suppressive action of CD8+ T-lymphocytes. In RCS/Cy compared to untreated tumor-bearing (RCS5) mice we report marked in vivo decreases in: (1) the activation (CD44HI/CD45RBLO phenotype) and proliferation (S + G2M phases of the cell cycle) of CD4+ T lymphocytes; (2) the ratio of activated and/or proliferating CD4+ to CD8+ T lymphocytes, and; (3) the proliferation of tumor cells. Also, depletion of CD8+ T lymphocytes from RCS/Cy mice abrogated much of the efficacy of the RCS/Cy treatment and led to changes in lymphoid populations more reminiscent of those in RCS5 than RCS/Cy mice. The data support our hypothesis that the RCS/Cy treatment achieves its efficacy by preventing the predominance of CD4+ over CD8+ T lymphocytes which is essential to maximum tumor growth in RCS5 mice. The results imply that analogous B-cell lymphomas in humans also may be treatable by shifting the T-cell balance toward inhibitory CD8+ rather than the tumor-stimulatory CD4+ T-lymphocytes. PMID- 8231238 TI - p145 expression during the cell cycle in HL-60 cell line and normal human lymphocytes: effects of camptothecin, vinblastine, cycloheximide, actinomycin D, retinoic acid and DMSO. AB - Bivariate flow cytometric analysis of nucleolar antigen p145 was performed on quiescent and phytohemagglutinin-stimulated human lymphocytes and on a promyelocytic cell line (HL-60). Data were acquired on a FACScan flow cytometer and analyzed using LYSYS II. Quiescent lymphocytes did not express p145. PHA stimulated proliferating lymphocytes expressed p145 maximally after 48 h, similarly to HL-60 cells in exponential growth. Antigen expression in G1 was notably heterogeneous in both cell types. The ratio of p145/DNA was highest in early S and decreased during mid and late S and remained low in G2M. p145 expression was lowest in M-phase cells treated for 6 h with vinblastine. Cycloheximide and actinomycin D had similar effects on p145 in HL-60 cells: expression of p145 gradually decreased from 1 to 6 h incubation in all phases of the cell cycle. Camptothecin did not decrease p145 expression and apoptotic cells from CAM-treated cultures still expressed p145. Retinoic acid and DMSO induced differentiation in HL-60 cells, and as this process progressed, p145 levels gradually fell until they approached isotype antibody control levels at 9 and 6 days, respectively. However, after 5 days treatment with 2 nM retinoic acid apoptotic cells appeared which still expressed p145. The data on drug treatment suggest that p145 exists in undifferentiated and proliferating cells and may not be a specific marker for malignancy, but may prove useful as a monitor of chemotherapeutic effects in cancer treatment. PMID- 8231239 TI - Minimal residual disease in murine B-cell leukemia (BCL1) detected by PCR. AB - Small proportions of leukemic cells escaping chemo-radiotherapy and/or dormant leukemic cells present in undetectable amounts in patients post therapy, i.e. minimal residual disease (MRD), are a source for relapse. The study of MRD and its treatment in a murine model for human B-cell leukemia/lymphoma, (BCL1), should lead to an improved understanding of the human disease. The standard assay for MRD in experimental mice is the adoptive transfer of spleen cells from experimental animals into naive secondary syngeneic recipients. We describe here the detection of MRD in BCL1-carrying BALB/c mice using the polymerase chain reaction (PCR). A BCL1 specific sequence from the rearranged VH-region was amplified yielding a 456 bp long fragment. PCR products hybridized to the cloned BCL1 sequence allowed the detection of a single BCL1 cell. This assay, therefore, reveals the presence of very small numbers of leukemic cells without sacrificing experimental animals. PMID- 8231240 TI - Linkage of proliferative and maturational abnormalities in chronic myelogenous leukemia and relevance to treatment. AB - Despite recent advances in our understanding of the molecular and biological abnormalities in chronic myelogenous leukemia (CML) this new knowledge has not yet led to significant improvements in treatment. We have reviewed what is known and still unknown about the molecular and biological abnormalities in CML that may be relevant to developing improved, more selective treatment. CML originates in a multipotential stem cell due to its acquiring a highly consistent specific chromosomal translocation between chromosomes 9 and 22; this results in a fused bcr/abl gene and an abnormal 210 kDa fusion protein which has increased intrinsic protein tyrosine kinase activity compared to the normal c-abl protein. It is still unknown how p210bcr-abl alters the signal transduction pathways, but the main biological abnormality is discordant or asynchronous maturation, with the cytoplasm generally maturing more rapidly than the nucleus. The major expansion of the CML population takes place in the intermediate and later maturation compartments rather than in the stem cell or early progenitor cell compartments. The expansion occurs slowly, probably taking several years to reach a trillion or more cells, at which time clinical symptoms begin to develop. The maturing leukemic progenitors do not have an increased proliferative rate, but they undergo one or more additional divisions and also live longer than comparable normal progenitors. The earliest CML blast cell population we have been able to study has reduced ultimate proliferative capacity compared to a comparable primitive normal blast cell population. Although no quantitative stem cell assay is available, indirect evidence suggests that the CML stem cells' biological behavior may be relatively unaffected or deviate only slightly from normal. The bcr/abl gene and its fusion protein are promising targets for development of novel specific therapies, but before this can be accomplished it will be necessary to understand more completely the molecular and biochemical abnormalities and to correlate them with the biological manifestations of the disease. PMID- 8231241 TI - Multi-institutional study of all-trans-retinoic acid as a differentiation therapy of refractory acute promyelocytic leukemia. Leukaemia Study Group of the Ministry of Health and Welfare. AB - We treated 70 acute promyelocytic leukemia (APL) patients with daily oral 45 mg/m2 all-trans-retinoic acid (ATRA) in two multi-institutional prospective studies. Of 64 evaluable patients, 21 were refractory to initial induction chemotherapy; 10 were refractory to salvage chemotherapy; 17, five, and four were in the first, second and, third relapse, respectively; and seven were previously untreated due to old age. In the first study with ATRA from China, 18 out of 22 (82%) evaluable patients achieved complete remission (CR). Initial peripheral leukemia cell counts were significantly less in the CR cases (p < 0.01); < 100/microliters in 17 out of 18 CR cases, and > or = 200/microliters in all failure cases. In the second study with ATRA from Hoffmann-La Roche, if initial leukemia cell counts were more than 200/microliters, chemotherapy was first given and then ATRA was started. Of 42 evaluable patients, 36 (86%) achieved CR. Morphological evidence of differentiation was noted in all CR cases. Patients achieving CR received standard consolidation and maintenance chemotherapies, and the 20-month predicted disease-free survival rate is 76% for cases achieving their first CR with ATRA. Toxicities attributable to ATRA were minimal and included cheilitis, xerosis, dermatitis, gastrointestinal disorders, bone pain, liver damage, and high serum triglyceridemia. PMID- 8231242 TI - Restoration of impaired cytokine secretion from monocytes of patients with myelodysplastic syndromes after in vivo treatment with GM-CSF or IL-3. AB - Cytokine treatment in patients with myelodysplastic syndrome (MDS) aims to overcome the maturation defects of myeloid lineage cells associated with cytopenia and cellular dysfunction of mature cells. Since phagocytes play a major role in host defense against microbial infection, we investigated cytokine secretion and oxygen radical release (ORR) from peripheral blood monocytes (PBMC) in a total of 16 MDS patients, 12 patients with refractory anemia (RA) and four patients with RA and excess of blasts (RAEB). Interleukin (IL-6), tumour necrosis factor alpha (TNF alpha), IL-1 beta, and IL-8 secretion from monocytes in response to lipopolysaccharide (LPS) was significantly reduced in the 12 patients with RA compared to 12 healthy controls, whereas no difference was seen in ORR. We further assessed cytokine secretion from monocytes of 10 MDS patients before and after therapy with granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-3, or a combination therapy with GM-CSF and cytosine arabinoside (AraC). In all 10 patients, secretion of IL-1 beta, IL-6, and TNF alpha from PBMC increased after cytokine therapy, whereas IL-8 secretion increased only in five patients with GM-CSF or IL-3 therapy receiving a dosage > or = 250 micrograms/m2 per day but decreased in all other patients. ORR increased in all patients on either GM CSF or IL-3 therapy. These data indicate that the ability of monocytes to secrete secondary cytokines is impaired in MDS patients but can be restored by in vivo administration of GM-CSF and IL-3. PMID- 8231243 TI - Toxicity following concurrent intrathecal and moderate-dose intravenous methotrexate. AB - To evaluate factors predisposing children with non-Hodgkin's lymphoma to toxicity from moderate dose methotrexate (MTX) (300 mg/m2 per 4 hours), we reviewed the medical records of 15 patients treated at our institution according to two similar protocols. Five patients experienced hyperemesis and/or severe mucositis. In two of these patients, pharmacokinetic analysis demonstrated delayed terminal excretion of methotrexate with a half-life of 3-3.5 days, compared to a previously reported t1/2 of 8-15 hours in subjects with normal clearance. All affected patients were large (body surface area 1.6-1.9 m2), and MTX toxicity was seen only during courses where intravenous MTX was given concurrently with intrathecal MTX. Four patients also received simultaneous prophylactic doses of oral trimethoprim-sulfamethoxazole (trimethoprim 5 mg/kg per day). We recommend that, in protocol design, consideration be given to avoiding concurrent use of intravenous and intrathecal MTX, and possibly trimethoprim-sulfamethoxazole. Where high doses of MTX are given based on large body surface area, urine alkalinization may be indicated. PMID- 8231244 TI - Frequent detection of BCR-ABL specific mRNA in patients with chronic myeloid leukemia (CML) following allogeneic and syngeneic bone marrow transplantation (BMT). AB - We performed a two-step polymerase chain reaction (PCR) to detect bcr-abl specific mRNA in 440 peripheral blood and/or bone marrow samples of 30 chronic myeloid leukemia (CML) patients (mean 15, range 2-50 samples) following non T cell-depleted allogeneic (n = 28) or syngeneic (n = 2) bone marrow transplantation (BMT). Median follow-up after BMT is 40 months (range 2-116 months), the median observation time 29 months (range 2-40 months). In 15 patients (50%), bcr-abl-specific mRNA could be detected following BMT. Bcr-abl positivity was rare in patients who were in hematological remission for at least 40 months (2/11). In five patients, PCR positivity was observed only once; all five patients are in complete hematological remission. Ten patients showed bcr abl specific mRNA in two or more consecutive samples. Hematological relapse occurred in five of the latter patients. Bcr-abl positivity preceded hematological relapse in all cases. Bcr-abl positivity was detected more frequently in patients without graft-versus-host disease (GVHD) (11/15), than in patients with GVHD (4/15) (p < 0.02). Our data indicate that transient bcr-abl positivity is not usually followed by hematological relapse, while patients, who are positive in serial samples have a high risk of relapse. PMID- 8231245 TI - The clinical correlations of serum tumor necrosis factor-alpha in acute leukemias: a predictor of response and relapse? AB - The clinical correlations of serum tumor necrosis factor alpha (TNF-alpha), a cytokine which can be released from leukemic blasts, has not been extensively studied. We have analyzed serum TNF-alpha in 20 ANLL, one CML-myeloblastic crisis, and 14 ALL adult patients by using a commercial ELISA kit. Sterile serum samples were taken on day 0, day 7, during remission and relapse with a mean follow-up period of 4.2 (1-19) months. After a median of 7 days following chemotherapy, serum TNF-alpha decreased both in responding ANLL (p = 0.004) and ALL (p > 0.05) but remained high in refractory leukemias. Values on day 7 were significantly different between responding and refractory patients in both ANLL (p = 0.0027) and ALL (p = 0.0099). At relapse, serum TNF-alpha increased starting at a median of 3 months preceding clinical symptoms in ANLL (p = 0.002). However, the relapse of ALL coincided with a slight increase which was not significant (p > 0.05). Together these findings indicate that serum TNF-alpha can be used as an early predictor of clinical response and relapse in ANLL. PMID- 8231246 TI - Quantitative and qualitative variation of ETS-1 transcripts in hematologic malignancies. AB - The ETS family proteins have a conserved DNA-binding domain and act as transcription factors. Three domains have been recently defined in human ETS-1 proteins and their role could depend upon the nature of alternative transcripts according to whether they possess or lack DNA binding and/or transcriptional activation domain and also point mutation that could affect these important domains. Expression of ETS-1 gene is very complex and is controlled at several levels: the initiation of transcription, alternative splicing, post-translational modification, and protein stability. As a selection apparently exists for ETS-1 gene activation in hematopoietic cells, we investigated a relation between quantitative and qualitative ETS-1 expression and leukemogenesis. Using Northern blot, polymerase chain reaction (PCR), and single strand conformation polymorphism (SSCP) methods, we analyzed quantitative and qualitative ETS-1 expression in a variety of hematological pathologies and cell lines of different origin. Two ETS-1 transcripts of 6.8 and 2.7 kb, resulting from differential polyadenylation site utilization and exhibiting different stability, were observed. We identified, in a great number of patients, the four alternative ETS 1 products, but the relative extent significance of the four transcripts was very different from one patient to another. A non-conservative mutation observed in one case of T-cell acute lymphoblastic leukemia (T-ALL) and in the ETS-1 transactivation domain raised the question of suppressor activity for some ETS-1 products, as it is now known that activators and repressors can be encoded by the same gene and consistently co-expressed in vivo. PMID- 8231247 TI - In vitro and in vivo effects of synthetic ribozymes targeted against BCR/ABL mRNA. AB - Chronic myelogenous leukemia (CML) is associated with a translocation of the BCR and the ABL genes, t(9;22). Results of this event are transcription and translation products that are unique to malignant cells. We therefore designed synthetic ribozymes which are capable of exclusively cleaving the BCR/ABL B3A2 type mRNA without altering normal cellular transcripts. Synthetic B3A2-type transcripts could only be cleaved by B3A2-type mRNA targeted ribozymes and not by any of the controls. The B3A2-type mRNA directed ribozyme, on the other hand, did not cleave any of the control transcripts. The effective delivery of ribonucleic acids by lipofection into K562 cells could be demonstrated by fluorescent microscopy, slot blot analysis, and RNA polyacrylamide gel electrophoresis. In vivo, we were able to induce a significant inhibition of the proliferation of K562 cells with ribozymes directed against the B3A2-type mRNA. Quantitative PCR analyses showed an up to fivefold reduction of the relative number of BCR/ABL mRNA molecules per single cell after exposure to ribozymes compared to controls. We conclude that ribozymes targeted against the B3A2-type BCR/ABL mRNA function in vitro and in vivo. PMID- 8231248 TI - Efficacy of acetyldinaline for treatment of minimal residual disease (MRD): preclinical studies in the BNML rat model for human acute myelocytic leukemia. AB - The efficacy of acetyldinaline [4-acetylamino-N-(2'-aminophenyl)-benzamide] for eradication of minimal residual disease (MRD), which is left after bone marrow transplantation, and the risk of a bone marrow graft being jeopardized by this treatment was studied in the Brown Norway rat acute myelocytic leukemia model (BNML). To mimic the clinical situation, MRD induction treatment was given to rats showing clinical signs of leukemia and consisted of 80 mg/kg cyclophosphamide and 7.0 Gy X-rays total body irradiation resulting in a 6-8 log leukemic cell kill leaving 10-1000 leukemic cells in the animals. Treatment was completed with a syngeneic bone marrow transplant. A high dose level (HD) treatment of 23.7 mg acetyldinaline/kg per day and a low dose level (LD) treatment of 11.85 mg/kg per day, each given orally for five consecutive days, were compared. The increase in the survival time, the cure rate, and the toxic death rate were evaluated. One 5-day course of LD treatment, started at a time interval of 10, 17, or 24 days following MRD induction, resulted in 44%, 11% or 0% cures. With two 5-day courses of LD treatment, 89%, 22%, or 0% cures were achieved. With LD treatment, maximally an 8 log leukemic cell kill was obtained and no toxicity-related deaths were observed (only less than a 1 log kill of normal hemopoietic stem cells). In contrast, a single course of HD treatment resulted in 56% of the rats (10/18) dying from intestinal tract toxicity, while from the remaining eight rats at risk for relapse, three (37%) showed a very late relapse and five were cured (63%). It was evident that the leukemic cell load at the start of the acetyldinaline treatment determined the probability of relapse. An important finding was that acetyldinaline did not interfere with bone marrow regeneration. The highly curative potential of acetyldinaline treatment in the BNML model during the phase of MRD warrants the introduction of this compound in clinical phase I/II studies. PMID- 8231249 TI - Regulation of B-cell lymphoma growth in syngeneic SJL/J mice. establishment of tumor dormancy following administration of anti-CD4 monoclonal antibody into tumor-bearing mice. AB - The spontaneously arising B-cell lymphoma (la+ reticulum cell sarcoma, RCS) in SJL/J mice has been shown to depend on host CD4 T cells for proliferation and growth. Treatment of mice with CD4 monoclonal antibody (mAb) prior to or after inoculation of a lethal dose of RCS tumor cells inhibits cell growth and the mice survive. The mechanism of tumor growth inhibition was studied by adoptively transferring cells from CD4 mAb treated tumor bearers into naive syngeneic mice. The recipient mice developed tumors and died. Tumor growth was dependent on the concentration of the adoptively transferred tumor cells. These results suggested that a state of tumor dormancy was established in the treated mice. Tumor dormancy was long lasting, as cells transferred as late as 11 weeks after the initial RCS inoculation still developed tumors in the recipient. The maintenance of dormancy was not due to failure of the recipient mice to mount an anti-tumor response or to the induction of suppressor cells. These results suggest that in the absence of CD4 cells, the RCS tumor remains dormant and this state of dormancy persists for long periods after total recovery of the CD4 cell subpopulation. Thus, it appears that RCS proliferation and growth is dependent on host CD4 cells but maintenance is under the influence of other cells or factors. Further characterization of this tumor dormant system and its regulation by the host may reveal novel mechanisms of tumor dormancy. PMID- 8231250 TI - Down-regulation of c-myc gene is not obligatory for growth inhibition and differentiation of human myeloid leukemia cells. AB - Suppression of c-myc expression is observed during induced differentiation of several myeloid cell lines and it has been attributed to the cell growth arrest that accompanies terminal differentiation. To dissect the role of c-Myc in the proliferation-differentiation switch we have studied c-myc expression in K562 cells exposed to several chemical agents. This model system allowed us to discriminate between the growth arrest and differentiation phenomena as well as the induction of differentiation along two different lineages (erythroid and myelomonocytic). Our results showed that c-myc expression did not significantly decrease when growth inhibition is reversible, either by treatment with a differentiating agent such as hydroxyurea (which induced erythroid differentiation) or by a non-differentiating agent such as interferon-alpha. In contrast, c-myc expression decreased when cells underwent terminal differentiation, either along the myelomonocytic (by 12-O-tetradecanoylphorbol-13 acetate) or erythroid (by 1-beta-D-arabinofuranosylcytosine) lineages. These results indicated that c-myc down-regulation is not obligatory for growth arrest and non-terminal differentiation of human myeloid cells. In contrast, c-myc down regulation occurred in terminal differentiation, but induction of myelomonocytic differentiation resulted in a greater loss of c-myc mRNA than induction of erythroid differentiation. PMID- 8231251 TI - The abnormal p53 proteins expressed in CML cell lines are non-functional. AB - Inactivation of the tumor suppressor function of the p53 gene is found in association with 20-40% cases of chronic myeloid leukemia (CML) in blast crisis. A common mechanism of p53 inactivation in CML is by complete deletion of one p53 allele in association with a point mutation which produces a mutant p53 protein on the remaining allele. Whether the mutant p53 protein, which is generally expressed at an elevated level, plays any role in the pathogenesis of blastic transformation or in maintaining the neoplastic proliferation, as it does in some solid tumors, is unknown. By using an antisense oligonucleotide approach, we investigated the cellular function of known abnormal forms of p53 protein, both mutant and truncated, expressed in CML cell lines. We found that the introduction of p53 antisense oligonucleotides can specifically inhibit the translation of the p53 mRNA. However, inhibiting p53 expression had no effect on cell proliferation, cell viability, and colony formation. There was no change in cell doubling time when the cells were maintained in serum-free medium (SFM) in the presence of antisense oligonucleotides compared with cells maintained in SFM alone. We conclude that the mutant or truncated p53 proteins expressed in the blast cells of CML have no growth-promoting effect and are not required for cell survival and proliferation. We further speculate that the loss of the tumor suppressor function of p53 might be the only mechanism by which p53 is involved in the transition from chronic phase to blast crisis. PMID- 8231252 TI - Phorbol ester induces apoptosis in HL-60 promyelocytic leukemia cells but not in HL-60 PET mutant. AB - One of the factors regulating the population size of a clone of proliferating cells is the induction of a physiological suicide mechanism known as apoptosis. We studied apoptosis in the HL-60 human promyelocytic leukemia cell line which differentiates when exposed to phorbol ester (S-cell), and in the PET-cell mutant of HL-60 which is defective in its response to phorbol ester. Exposing S-cells to 12-O-tetradecanoylphorbol 13-acetate (TPA) (3 nM and above) induced morphological changes characteristic of apoptosis (visualized by light microscopy), and induced fragmentation of chromatin DNA to oligonucleosomal lengths. These changes were obvious in 48 h. In contrast, 1000 nM TPA for five days did not induce apoptosis in the PET-cell. DNA fragmentation was induced in both cell lines by A23187 (0.25 microM) and etoposide (7 microM). Novobiocin (600 and 900 microM) induced DNA fragmentation in S-cells, but higher concentrations inhibited fragmentation. Novobiocin is believed to induce DNA fragmentation by a direct action on DNA. In the case of PET-cells, novobiocin did not induce DNA fragmentation at any concentration, and prior treatment of PET-cells with novobiocin (300-1200 microM for 30 min) inhibited DNA fragmentation induced by A23187. Novobiocin inhibited cell growth equally in S-cell and PET-cells. It is concluded that the promyelocytes have the capacity to undergo apoptosis in response to agents which activate protein kinase C, and that the PET-cell has a mutation which disables both protein-kinase C-induced and novobiocin-induced DNA fragmentation, leaving intact the ability of novobiocin to protect DNA from calcium-entry-initiated fragmentation. The elucidation of the lesion responsible for the PET phenotype is likely to increase our understanding of this important pathway for regulating cellular proliferation and how it bears on leukemogenesis and chemotherapy. PMID- 8231253 TI - Infection of leukaemic B lymphocytes by Epstein Barr virus. AB - Epstein-Barr virus (EBV) infection of B lymphocytes in vitro gives rise to immortalized lymphoblastoid cell lines. Previous reports have shown that chronic lymphocytic leukaemia (CLL) cells, although infectable by EBV, are resistant to immortalization (1-4), although a small number of CLL cell lines have been reported (5-7). In the present study we have analysed early events occurring after EBV infection in 16 CLL samples. Out of 16 samples, 15 could be infected by the virus and expressed the full EB viral nuclear antigen (EBNA) complex but only one out of 16 expressed the latent membrane protein (LMP). The five CLLs in which we could investigate the presence of viral episomes showed circularized EBV by 16 hours after infection. The sequence of EBNA expression and genome circularization mirrored that seen in normal B cells, although genome amplification was not detected. The only CLL sample which expressed LMP after EBV infection was induced to proliferate for 2-3 weeks, but no cell line was established. Immortalized cell lines were obtained from three out of 16 samples tested, but all were polyclonal for light chain expression and had arisen from the CD5-negative, normal B-cell population. Thus the inability of EBV to induce proliferation of most CLL cells correlated with the absence of LMP expression which is invariably expressed during immortalization of normal B cells. This novel type of restricted gene expression could be compatible with evasion of host immune responses and consequent long-term survival of the cell in vivo. PMID- 8231254 TI - Establishment of cell lines from B-cell precursor acute lymphoblastic leukemia. AB - In an attempt to establish permanent cell lines from children with B-cell precursor acute lymphoblastic leukemia (BCP-ALL), 123 clinical samples from 117 patients were cultured in vitro. Using a method which was successful for the growth of ALL with T-cell phenotype, 3% (2/74) of BCP-ALL samples from patients at diagnosis and 31% (9/29) of BCP-ALL samples from patients at relapse were established as cell lines. However, in most cultures, leukemic cells survived for only a few weeks and the majority of viable cells present after 28 days of culture were esterase-positive mononuclear cells. Based on the hypothesis that mononuclear cells inhibited leukemic cell growth, we evaluated the effect of a monocyte toxin, L-leucine methyl ester (Leu-OMe), on the growth of four frozen BCP-ALL samples. Thawed leukemic cells treated with Leu-OMe, but not untreated control cells, proliferated in three samples and one new cell line was established. Subsequently, when Leu-OMe was added to fresh leukemia cells in culture, leukemic cell lines were grown from 29% (4/14) of samples at diagnosis and 66% (4/6) of relapse samples. Overall, 20 BCP-ALL cell lines were established, all were Epstein-Barr virus (EBV)-negative, and authenticity of each cell line was verified by a direct comparison of the immunophenotype, karyotype, and genotype with the patient's tumor cells. This improved method of cell culture permits a higher success rate of cell line establishment from patients with BCP ALL thereby aiding in analysis of B-lymphocyte transformation and neoplasia. PMID- 8231255 TI - Time-space case clusters of Burkitt's lymphoma in Malawi. AB - The geographical co-ordinates of 146 cases of Burkitt's lymphoma in Malawi, with date of onset between July 1987 and October 1989, were recorded. Case clusters, pairs of cases, closer together in time and space than would be expected by chance, were discovered, using Knox's method, for children over the age of 8 years, but not for all ages. PMID- 8231256 TI - Detection of circulating tumor cells in multiple myeloma by a PCR-based method. AB - Multiple myeloma (MM) is a B-cell malignancy characterized by clonal expansion of plasma cells producing monoclonal immunoglobulins. It has been regarded as a tumor typically involving only the bone marrow. The existence of circulating tumor cells has been suggested from phenotypic and genotypic studies. However, this issue is still controversial due to the limitations of the methods so far used. We describe a novel polymerase chain reaction (PCR) based method using clone-specific immunoglobulin heavy-chain gene sequences as tumor markers. From such sequences patient-specific oligonucleotide primers and probes were generated, and used to detect tumor cells. Seven MM patients were selected for this study, and tumor cells were found in all peripheral blood samples. The demonstration of circulating tumor cells suggests some caution when using peripheral blood for autograft procedures, even though its contamination is lower than bone marrow. In conclusion, we describe a specific and sensitive PCR-based method for detecting minimal disease which is of general applicability to all lymphoid malignancies transcribing rearranged immunoglobulin heavy-chain genes. PMID- 8231257 TI - The chronic myelocytic cell line K 562 contains minor (m) as well as major (M) ber/abl fusion mRNAs. AB - By searching for additional chimeric bcr/abl transcripts in K 562 cells characterized by major (M) bcr/abl fusions, a new mRNA, a minor (m) bcr/abl transcript, was detected. A practical implication of this finding is that the K 562 cell line can be used as positive control for the detection by the polymerase chain reaction of both types of transcripts for the diagnosis of Philadelphia chromosome associated leukemias. PMID- 8231258 TI - Use of a retinoblastoma gene probe to investigate clonality in Richter's syndrome. AB - A 69-year-old woman presented with Rai stage 0 chronic lymphocytic leukemia. Ten years later she developed a diffuse centroblastic lymphoma involving the stomach. The surface membrane phenotype of the CLL cells was MD lambda while that of the large cell lymphoma (LCL) cells was MD kappa. The two populations had different heavy and kappa light chain rearrangements. Cytogenetic analysis of the CLL cells showed a deletion involving chromosome 13, band q14, but was unsuccessful in the LCL cells. However, use of a probe (p68 RS2.0) which recognizes a variable number tandem repeat sequence in the retinoblastoma gene, localized to chromosome 13q14, showed two alleles in the LCL cells but only one in the CLL cells. These data suggest that in this case of Richter's syndrome the CLL cells and the LCL cells are clonally distinct. PMID- 8231259 TI - Occurrence of a Ki-1-positive anaplastic large-cell lymphoma in a patient with Ph' positive chronic myelogenous leukemia successfully treated by alpha interferon. AB - We describe a patient with Philadelphia-chromosome-positive (Ph' +) chronic myelogenous leukemia (CML), who developed an anaplastic large cell lymphoma (ALCL) with T-phenotype, after 43 months successful treatment with alpha interferon (IFN). Characterization studies of lymphoma cells showed positivity for Ki-1 monoclonal antibody, T-cell surface markers, T-cell receptor beta chain rearrangement, and germline configuration of the BCR gene. At the time of lymphoma diagnosis, the patient had achieved complete hematologic remission from CML with partial karyotypic conversion (50% Ph' + cells). After twelve weekly courses of polychemotherapy, he obtained complete remission from lymphoma. At present, five years from CML diagnosis, the patient has a remarkably stable disease, being in remission from lymphoma and in well controlled CML chronic phase. Our case thus represents the first well documented description of a T-cell non-Hodgkin's lymphoma developed during the course of CML. PMID- 8231260 TI - A case of acute megakaryocytic leukemia with hematogones. AB - A case of an infant male with Down's syndrome, pancytopenia, and a relative lymphocytosis is presented. A bone marrow aspirate and crushed marrow biopsy were evaluated by microscopy and flow cytometry. A population of immature lymphoid cells positive for CD10 (CALLA) and CD19 (Leu 12) was initially interpreted as suggestive of an acute lymphoblastic leukemia (ALL). However, a second bone marrow biopsy revealed morphologic features consistent with acute megakaryocytic leukemia (AMKL) with fibrosis. This diagnosis was supported by the flow cytometric study showing increased myeloid and megakaryocytic antigen-positive cells in the bone marrow and peripheral blood obtained after chemotherapy for ALL. This report presents evidence that the original lymphoid population represented hematogones, which were preferentially aspirated due to marrow fibrosis. There are no previous reports of hematogones identified at the presentation of an acute leukemia. It also emphasizes that the presence of hematogones with the resultant marker findings can be misleading. The diagnosis of ALL in pediatric patients should not be based solely on the presence of increased CD10-positive cells. PMID- 8231261 TI - Adult T-cell leukemia/lymphoma revealed by a surgically cured cardiac valve lymphomatous involvement in an Iranian woman: clinical, immunopathological and viromolecular studies. AB - A 60-year-old woman from the town of Mashhad in northeastern Iran developed cardiac failure due to aortic and mitral regurgitations which needed cardiac valve replacement. Histopathological study of the valves revealed a T-cell non Hodgkin's lymphoma. Blood examination showed leukemic features with 32% of abnormal white blood cells. Human T-cell leukemia/lymphoma virus type I (HTLV-I) antibodies were present in the serum and the specific env HTLV-I sequences were detected in the DNA extracted from the valves and peripheral blood mononuclear cells (PBMC) using polymerase chain reaction technique. Clonal integration of two HTLV-I copies was found in both the valves and PBMC DNA, thus the diagnosis of adult T-cell leukemia/lymphoma (ATL) was established. In contrast to the acute life-threatening cardiac localization, our case met the diagnostic criteria of chronic ATL, this was confirmed by favorable evolution without chemotherapy during the 24 months after diagnosis. According to our knowledge, this is the first report of an isolated lymphomatous cardiac valve involvement, without other cardiac abnormalities. It seems important to underline that the patient originated from Iran where endemicity of HTLV-I has only recently been discovered. PMID- 8231262 TI - Classical staging of Hodgkin's disease is inappropriate for selecting patients for clinical trials of intensive therapy: the case for the objective use of prognostic factor information in addition to classical staging. PMID- 8231263 TI - Prognostic factor studies in Hodgkin's disease: problems and pitfalls. PMID- 8231264 TI - Two types of ring chromosomes found in a patient with myelodysplastic syndrome. PMID- 8231265 TI - Fluorescent in situ hybridization analysis of an atypical t(11;14)(q13;q32) without expression of the cyclin D1 gene. PMID- 8231266 TI - 75th anniversary of Mayo dietetics. PMID- 8231267 TI - Identification of a polymorphism in the human neurotensin receptor gene. AB - A complementary DNA (cDNA) clone encoding the neurotensin receptor was isolated from a human substantia nigra cDNA library. The deduced amino acid sequence of this clone was almost identical to that of a cDNA for this receptor cloned from a previously described HT29 human colonic adenocarcinoma cell line. We found three base changes between the previously reported HT29 cDNA clone and the current cDNA clone. We investigated these changes by using polymerase chain reactions to amplify these areas from various human samples. One of the differences, which resulted in an amino acid change at AA194 (a leucine in the HT29 sequence was a phenylalanine in the current sequence), was found in some, but not in all, human samples. This finding represents genetic variability in human neurotensin receptors, the first such report for a peptide receptor. Both of these receptors, however, when expressed separately in transfected cell lines, had similar affinities for neurotensin and some related peptides. PMID- 8231268 TI - Immunohistochemical characterization of hepatic lymphocytes in primary biliary cirrhosis in comparison with primary sclerosing cholangitis and autoimmune chronic active hepatitis. AB - We analyzed the immunophenotypes of hepatic cellular infiltrates by quantitative immunohistochemical methods in biopsy specimens from 20 patients with primary biliary cirrhosis (PBC), 19 with primary sclerosing cholangitis, and 11 with autoimmune chronic active hepatitis. Specifically, we sought to identify activated T cells, interferon-gamma-producing cells, and natural killer cells. The portal cellular infiltrate in PBC contained a preponderance of CD4 cells in comparison with CD8 cells, with a CD4/CD8 ratio of 2.45:1. The cellular infiltrate in areas of piecemeal necrosis contained mostly CD8 cells. Infiltrating CD8 cells in PBC had the surface phenotype of cytotoxic (CD8 positive, CD11b-negative) cells. Approximately 4% of T cells expressed interleukin 2 receptors. Interferon-gamma-staining cells were rarely identified (in less than 2%). Cells that expressed the natural killer cell markers CD16, CD56, or CD57 were infrequent, constituting approximately 5% of the cellular infiltrate. The composition of the infiltrates was similar in patients with PBC and chronic active hepatitis. Natural killer cells were twice as common in patients with primary sclerosing cholangitis (P < 0.05) as in those with PBC. The inflammatory infiltrates in areas of piecemeal necrosis were similar in the three diseases and differed from those found within the portal area, in that CD8 cells were preponderant. In all three liver diseases, almost 90% of bile ducts expressed class II HLA antigens. These findings support the hypothesis that cytotoxic T cells of either the CD4 or CD8 immunophenotype but not natural killer cells may be involved in the pathogenesis of PBC and chronic active hepatitis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231269 TI - Surgical treatment of vascular rings: the Mayo Clinic experience. AB - From 1947 through 1992, 37 Mayo Clinic patients underwent operation for the relief of tracheoesophageal obstruction that resulted from vascular rings and related entities. Of the 37 patients, 18 had a double aortic arch, 11 had a right aortic arch with an aberrant left subclavian artery, 4 had a left aortic arch with an aberrant right subclavian artery, 2 had a pulmonary artery sling, 1 had a right aortic arch with mirror-image branching and a left ligamentum arteriosum, and 1 had a left aortic arch, a right descending aorta, and a right ductus arteriosus. Symptoms consisted of stridor, recurrent respiratory infections, and dysphagia. The anomaly was approached through a left thoracotomy in 31 patients, through a right thoracotomy in 4, and through a median sternotomy in 2. Only one early postoperative death (3%) and no late deaths occurred. At long-term follow up (maximal duration, 45 years), three patients had residual symptomatic tracheomalacia, one of whom required right middle and lower lobectomy for recurrent pneumonia. Magnetic resonance imaging is the imaging technique of choice for accurate delineation of the vascular and tracheal anatomy. When patients are symptomatic, vascular ring should be repaired. The surgical risk is minimal, and the long-term results are excellent. PMID- 8231270 TI - Laparoscopic placement of ventriculoperitoneal shunts: preliminary report. AB - We used a laparoscopic technique for the percutaneous placement of the peritoneal end of cerebrospinal fluid shunts in adult patients with obstructive or normal pressure hydrocephalus. Concurrent with the initial cranial part of the procedure, pneumoperitoneum is established in a routine fashion, and a video laparoscope and grasping forceps are inserted into the peritoneal cavity. With use of a pacemaker introducer kit, the peritoneal catheter is placed percutaneously under direct laparoscopic vision through a small upper abdominal incision into the peritoneal cavity. At the completion of the procedure, the patency of the assembled shunt system can be verified by observing free flow of cerebrospinal fluid from the catheter tip as the valve is being pumped. We found that this technique is particularly useful in technically challenging cases--for example, those involving obese patients and those who have undergone multiple abdominal operations. No complications associated with the technique were encountered. PMID- 8231271 TI - Achalasia: prospective evaluation of relationship between lower esophageal sphincter pressure, esophageal transit, and esophageal diameter and symptoms in response to pneumatic dilation. AB - The aims of this study were to investigate a group of patients with achalasia prospectively to determine (1) the relationship between changes in symptoms and esophageal motor function in response to pneumatic dilation and (2) the effects of the balloon size as well as the frequency and duration of inflation on the outcome of treatment. Fourteen patients with achalasia who were symptomatic for a median duration of 27 months participated in the study. The patients were randomized to one combination of the following pneumatic dilation conditions: a 30- or 35-mm balloon dilator, one or two balloon inflations, and 20, 40, or 60 seconds per balloon inflation. A comprehensive assessment of their symptoms and esophageal motility, transit, and diameter were performed before and 3 months after pneumatic dilation. Pneumatic dilation provided significant relief of dysphagia (P < 0.01), but other symptoms (heartburn, regurgitation, and chest pain) remained unchanged. Pneumatic dilation also caused a significant decrease in lower esophageal sphincter pressure and esophageal diameter and improved esophageal emptying of a solid bolus. Nevertheless, no significant association was detected between changes in the symptom score for dysphagia and changes in objective response measures as a result of pneumatic dilation. Changes in the symptom score for dysphagia or objective responses were similar regardless of the size of the dilator used or the frequency and duration of the balloon inflations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231272 TI - Management of orbital tumors. AB - Orbital tumors are uncommon. In children, both malignant and benign causes of orbital proptosis necessitate urgent assessment; in many cases, emergent intervention is necessary to avoid blindness. In adults, proptosis is most commonly associated with thyroid orbitopathy. Orbital tumors in adults rarely are characterized by the explosive growth and damage that can occur with childhood lesions. In both age-groups, the evolution of better scanning modalities, such as magnetic resonance imaging with fat saturation and gadolinium enhancement, has improved diagnostic accuracy, especially in patients with loss of vision. In more than 95% of cases, noninvasive techniques yield a correct diagnosis. In patients who require nonsurgical intervention, especially if the diagnosis is uncertain, fine-needle aspiration biopsy has an accuracy that exceeds 95%. Combined-modality therapy has improved the control of and decreased the morbidity associated with several orbital tumors. Surgical advances, such as the ancillary use of the CO2 laser, have enhanced the management of some orbital tumors. PMID- 8231273 TI - Primary lymphoma of the spinal cord. PMID- 8231274 TI - Thigh injuries in athletes. AB - The thigh, which consists of the heavily muscled region around the femur, is vulnerable to many types of athletic injury. This review addresses the assessment, prevention, and rehabilitation of both the common thigh injuries associated with participation in sports, such as contusions and myositis ossificans traumatica, and a few relatively uncommon but diagnostically important entities that sports physicians must recognize when an athlete has pain or dysfunction attributable to the thigh. Dividing the thigh into anatomic compartments aids in both differential diagnosis and understanding of the pathomechanics that lead to injury. Rehabilitation is especially emphasized because improper selection of modalities or misuse of exercise can seriously impede or worsen recovery. Conversely, the use of appropriate rehabilitation principles and new concepts in exercise prescription can decrease morbidity and lead to rapid resumption of sports. PMID- 8231275 TI - Serologic diagnosis of parvovirus B19 infections. PMID- 8231276 TI - Thymoma. AB - Thymomas and thymic carcinomas are thymic epithelial tumors that constitute approximately 15% of all mediastinal masses. From 28 to 66% of thymomas cause chest symptoms as the initial manifestation; the rest are discovered on routine chest roentgenograms or during investigations prompted by the presence of a paraneoplastic syndrome. Forty percent of patients with thymoma have one or more paraneoplastic syndromes, including myasthenia gravis, pure red cell aplasia, and hypogammaglobulinemia. Extrathymic malignant lesions develop in up to 20% of patients. Traditional histologic classifications have not accurately predicted tumor behavior; a recently developed classification based on cellular differentiation toward thymic medullary or cortical epithelium may correlate better with prognosis. Nevertheless, the prognosis is best predicted by stage of the tumor determined intraoperatively and is poorer in patients with incomplete resection than in those with complete resection of the thymoma. In addition to surgical intervention, irradiation and chemotherapy have important roles in the management of thymomas, particularly in advanced stages. In this article, the clinical manifestations, diagnosis, pathologic features, staging, and treatment of thymomas are reviewed, and the prognosis of affected patients is discussed. PMID- 8231277 TI - International normalized ratio and prothrombin time. PMID- 8231278 TI - International normalized ratio and prothrombin time. PMID- 8231279 TI - Anders Celsius--temperature scale. PMID- 8231280 TI - Diabetes control and complications trial: the continuing challenge ahead. PMID- 8231281 TI - Primary biliary cirrhosis: cells, sera, and soluble factors. PMID- 8231282 TI - Vascular rings, slings, and tracheal rings. PMID- 8231283 TI - Influence of aging on the beta- and glucagon-receptor-mediated glycogenolysis in rat hepatocytes. AB - The influence of aging on beta-receptor and glucagon-receptor control of glycogenolysis was investigated in rat hepatocytes. The beta-receptor-induced glucose output was detectable only in senescent rats, was partly dependent on extracellular Ca2+, and was inhibited by 4 beta-phorbol 12-myristate 13-acetate (PMA), insulin, and the Ca(2+)-antagonists, verapamil and nifedipine. Chelation of extracellular Ca2+ potentiated the effect of nifedipine only. In contrast, glucagon-stimulated glycogenolysis, similar in mature and senescent rats, was independent on extracellular Ca2+ and was unaffected by PMA. Verapamil, in senescent rats only, and nifedipine, in mature and senescent rats, inhibited glucagon-stimulated glucose output only in the presence of Ca2+. Insulin inhibited glucagon-induced glucose output, irrespective of the age of the rat and the presence of Ca2+. We conclude that the beta-receptor component in the adrenergic regulation of glycogenolysis in senescent rats consists of a major Ca(2+)-independent and a minor Ca(2+)-dependent part, displaying different sensitivity towards protein kinase C (PKC), Ca(2+)-antagonists, and insulin. Aging does not change the capacity of glucagon to induce a full glycogenolytic response in the absence of extracellular Ca2+; Ca(2+)-influx, however, seems to be involved when extracellular Ca2+ is present, and this sensitivity is increased on aging. PMID- 8231284 TI - Age-dependent changes in the phosphorylation of nuclear proteins of submandibular glands in isoproterenol-treated rats. AB - Age-dependent changes in the regulation of the phosphorylation of nuclear proteins were investigated in relation to DNA synthesis in rat submandibular glands after injection of isoproterenol (IPR). The level of phosphorylation of nuclear proteins in the tissue increased rapidly after birth, reaching a maximum at 4 weeks, and then decreased to the level of 52-week-old rats. The level of protein kinase activity in nuclei varied in parallel with that of phosphorylation of nuclear non-histone proteins after birth. The time after the injection of IPR required to initiate the phosphorylation of nuclear non-histone proteins of rat submandibular glands, which occurs prior to the onset of RNA synthesis that precedes the replication of DNA, increased with age. These results suggest that this delayed onset of phosphorylation plays a regulatory role in cell proliferation and cell function during aging. PMID- 8231285 TI - Age-related changes in albumin binding by renal brush-border membrane vesicles. AB - A selective proteinuria occurs with normal aging. We investigated the contribution of a defect in the receptor-mediated endocytosis to the age-related albuminuria by measuring albumin binding by renal brush-border membrane vesicles from young and old female Wistar rats using a filtration method. Old (24 months) rats had a significantly higher proteinuria (13.29 +/- 5.25 mg prot/24 h/100 g bw) than did young (3 months) rats (1.23 +/- 0.55 mg prot/24 h/100 g bw). Scatchard analysis of the kinetic parameters of 125I-albumin binding revealed a decrease in the binding capacity of brush-border membrane vesicles from old rats. The number of binding sites, N (pmol/mg protein/min) was 236.84 +/- 97.50 in old rat preparations and 380.27 +/- 178.36 in young rat vesicles (P < 0.05). By contrast, Km did not change significantly with age (478.86 +/- 259.29 nM in old rat vesicles and 498.00 +/- 220.36 nM in young rat preparations). Consequently the index of adsorptive endocytosis efficiency (the N/Km ratio) decreased drastically with age from 0.782 +/- 0.238 at 3 months to 0.547 +/- 0.199 at 24 months (P < 0.05). These data indicate that defective receptor-mediated endocytosis could, at least partly, explain the age-dependent rise in urinary albumin excretion. PMID- 8231286 TI - Effect of caloric restriction on aflatoxin B1-induced DNA synthesis, AFB1-DNA binding and cell proliferation in Fischer 344 rats. AB - Young adult male Fischer rats maintained on a reduced calorie diet (60% of ad libitum food consumption) for 6 weeks showed a decrease in the binding of aflatoxin B1 (AFB1) to hepatic or renal nuclear DNA and a reduction of AFB1 induced hepatocellular damage. Repeated dosing of rats with AFB1 resulted in the inhibition of hepatic and renal DNA synthesis measured by [3H]thymidine incorporation. However, the rate of DNA synthesis was greater in ad libitum (AL) rats than in calorically restricted (CR) animals. Three days after AFB1 dosing, the rate of DNA synthesis had recovered to the control level. Cell cycle analyses measured by a flow cytometric method on kidney cells of both AL and CR rats showed that there were no significant changes in cell populations in the S phase between these two groups of rats. AFB1 inhibited the cell proliferation on an average of 33%. The restoration of the cell proliferation in kidney cells was found on the third day after AFB1 dosing. The rate of the regenerative cell proliferation was found to be slightly greater in AL rats than in CR animals. The AFB1-induced regenerative DNA synthesis in both liver and kidney was retarded by CR. PMID- 8231287 TI - Paracrine stimulation of senescent satellite cell proliferation by factors released by muscle or myotubes from young mice. AB - The proliferative potential of satellite cells undergoes a dramatic decrease in the early postnatal period and a more modest but continuous decrease throughout the life span of the animal. To address the problem of the mechanism regulating this phenomenon and to understand whether it is causally linked to senile muscle atrophy, we studied the response of aged satellite cells to serum and to different growth factors. The data reported indicate a generalised reduction in the response to all mitogens tested, which could not be compensated for by increased concentrations of serum or growth factors. On the other hand, conditioned medium of differentiated myotubes from young mice exhibited a strong mitogenic action on aged satellite cells, while conditioned media of myotubes from old mice or from a variety of non-muscle cells were ineffective. Furthermore, saline extracts from muscle of young mice are also able to exert this mitogenic action. Saline extracts of muscle from old mice were poorly mitogenic for satellite cells from young mice, and not at all for satellite cells from old mice. These data indicate that paracrine interactions operate inside the muscle tissue and are probably required for the normal replicative behaviour of satellite cells. The failure of such interactions may be among the causes leading to age-related muscle hypotrophy. PMID- 8231288 TI - Impact of aging on the morphology and function of the somatotroph cell population in rats. AB - There is little information regarding the impact of aging on pituitary somatotroph (STH) cell population in rats. We therefore undertook a quantitative immunocytochemical assessment of this cell type in young (3 months), old (20 months) and senescent (29 months) male rats. An attempt was also made to correlate morphological parameters with serum levels of growth hormone (GH). Since thyroid status is highly influential on somatotropic function, serum levels of thyroxine (T4) and triiodothyronine (T3) were also measured in the three age groups. We found a marked age-related reduction in STH cell number, volume density, and surface density, as well as a milder but significant decline in STH cell area and perimeter. Basal serum levels of GH remained unchanged with age, whereas the estimated number and amplitude of GH pulses declined from young to old animals. Thyroxine but not T3 levels also declined with age. We conclude that in rats, aging causes a marked reduction in somatotroph number and, to a lesser extent, cell size. These alterations do not affect trough levels of circulating GH. Our data also suggest that the progressive hypothyroidism associated with aging in this species may contribute to the promotion of the above changes. The present study emphasizes the convenience of combining hormone measurements with quantitative morphological analysis of cell populations for the study of pituitary function during aging. PMID- 8231289 TI - Effect of age and of swimming-induced stress on the phagocytic capacity of peritoneal macrophages from mice. AB - The effect of age and physical activity stress (swimming until exhaustion) on the phagocytic capacity of the peritoneal macrophages was studied in young (12 +/- 4 week-old) and old (68 +/- 6-week-old) BALB/c mice. The attachment and ingestion of opsonized Candida albicans as well as the ingestion of inert particles (latex beads) by these cells was compared between young and old animals at rest. The results show that phagocytosis of peritoneal macrophages is not impaired in old mice. With respect to the effect of physical activity stress, we evaluated the phagocytic capacity both of C. albicans and of latex beads before and immediately after swimming until exhaustion, and in the absence or presence of a previous period (1 month) of training. The results indicate that the phagocytic capacity of peritoneal macrophages is increased after swimming until exhaustion, both with and without previous adaptation or training, and in both young and old mice. PMID- 8231290 TI - Age and GSH metabolism in rat cerebral cortex, as related to oxidative and energy parameters. AB - A comprehensive study on GSH metabolism in relation to some markers of oxidative and energy status in rat cerebral cortex as a function of age was performed. Reduced GSH, total GSH and the GSH Redox Index decreased both during growth (defined as the period between 1 and 5 months) and during aging (defined as the period between 5 and 27 months) while GSSG levels increased during the two periods, but most significantly during aging. Also GSH-associated enzymes and adenine-pyridine nucleotide levels show age characteristic changes. The obtained results suggest that decreases in oxidative and energy metabolism occur during aging. They probably contribute to decreases in the activity of the biosynthetic processes (i.e., NADP+(H) and GSH synthesis) and in the antioxidant capacity of the GSH system. However, the oxidative stress does not seem to be a typical characteristic of the aging period; as an oxidative status is present during the growth period too. Typical parameters of aging process are mainly the low levels of reduced GSH, total GSH and GSH Redox Index and the high levels of GSSG as well as the high levels of GSH peroxidase and GSH transferase and the low levels of gamma-glutamylcysteine synthetase. PMID- 8231291 TI - Reduced density of dopamine D2 receptors in the rat tail artery as a function of age. AB - The present study was designed to investigate the influence of aging on noradrenaline content and the density and pattern of prejunctional dopamine D2 receptors in the tail (ventral caudal) artery of male Sprague-Dawley rats. Tail artery is frequently used as a model for investigating mechanisms of sympathetic vascular control and contains prejunctional dopamine receptor belonging to the D2 subtype. Noradrenaline levels were reduced in rats of 12 months of age in comparison with 3-month-old animals. A further reduction in catecholamine concentration was found in 24-month-old rats. The density of prejunctional D2 receptors, which was measured in frozen sections of the tail artery by using both radioligand binding and autoradiographic techniques, was reduced by about 35% in 12-month rats in comparison with 3-month rats. A decrease by about 55% versus 3 month rats and by about 20% versus 12-month rats was observed in 24-month-old rats. Neither the pharmacological profile nor the anatomical localization of prejunctional D2 receptors was changed in the rat tail artery as a function of age. The parallel decrease in noradrenaline content and in the density of prejunctional dopamine D2 receptor sites in the tail artery of Sprague-Dawley rats of different ages probably accounts for the lack of an age-related change of prejunctional dopamine D2 receptor-linked responses reported in functional studies. PMID- 8231292 TI - Low risk to certain diseases in aging: role of the autonomic nervous system and calcium metabolism. AB - The low risk of aging Africans, as opposed to high risk of Caucasians, to certain major disorders, including Parkinson's disease, myocardial infarction, osteoporosis and fractures, some rheumatic diseases, and an overall reduced incidence of cancer, has not been explained. In this study it is proposed, firstly, that relative risk is determined by a common physiological mechanism in which ANS status and calcium metabolism play a central role; secondly, that distinctive features of this mechanism in Africans may be subtly increased vagal tone, relatively enhanced dopaminergic versus noradrenergic activity, and an efficient dopamine/vitamin D-parathormone, anabolic hormone regulation of bone metabolism, and cell calcium homeostasis; and thirdly, that the neuroendocrine metabolic context determines the response to specific stimuli; consequently, 'risk' factors, as defined for particular disorders, are not universally applicable. Maintained dopaminergic activity, as proposed for Africans, coupled with low risk to certain disorders, confirms the experimentally demonstrated paramount importance of this neurotransmitter in retarding aging processes in animals. The neuroendocrine profile as defined for Africans is consistent with a potentially extended period of physical and mental competence and a conceivable shorter duration of involutionary decline. PMID- 8231293 TI - Estimation of left ventricular myocardial elasticity and viscosity by a thick walled spherical model. AB - The authors measured the transfer function (TF) of the left ventricle (LV) in an isolated canine preparation. Here TF indicates the ratio of induced vibration in LV to input vibration when an external mechanical oscillation is applied. TF had a single peak the frequency of which changed from 40 Hz to 80 Hz when LV pressure (LVP) increased from 6 mm Hg to 96 mm Hg. A mathematical model was formulated to estimate the viscoelasticity of the spherical shell. This model was constructed of the material points, elastic components which connected all the material points, and viscous components placed in series with elastic components. Theoretical TF can be computed if the viscoelastic values are given. The value of viscoelasticity at which the theoretical TF best fitted the experimental TF was considered to be the viscoelasticity of the model. The validity of this approach was verified using a silicone spherical shell. The estimated myocardial elasticity was 40 kPa when LVP was 6 mm Hg, 160-170 kPa when LVP was 96 mm Hg and was approximately proportional to LVP, whereas viscosity showed small change. The inclination of elasticity was consistent with previous reports. These results proved that myocardial elasticity can be estimated by analysing the transfer function of the left ventricle. PMID- 8231294 TI - Detection of late potentials in the signal-averaged ECG combining time and frequency domain analysis. AB - The paper reports experience of flexible, computer-based instrumentation for high resolution ECG, designed for laboratory research and clinical applications. It is an attempt to improve methods of detection of late potentials when identifying patients at increased risk of ventricular tachycardia (VT) and sudden cardiac death. Several factors influencing the efficiency of a high-resolution ECG system are examined. Of primary importance are signal, gain, type of filter, length and position of the time interval taken for spectral analysis, type of window used etc. Some possible error sources are analysed. Time-domain analysis is illustrated by representative XYZ-vector magnitude tracings of a non-VT and a VT patient. In an effort to further refine the diagnostic strategy, an approach to data analysis is presented, combining time-domain with frequency-domain measurements. A spectral ratio is proposed that helps recognise patients prone to VT. Two-dimensional plots of this ratio against time-domain parameters reveal clearly distinguishable normal and pathologic groups. It can be concluded that an appropriate frequency analysis can add significant diagnostic power to existing instrumentation for signal-averaged ECG to predict VT risk. PMID- 8231295 TI - Doppler radar device as a useful tool to quantify the liveliness of the experimental animal. AB - The Doppler radar device which is described here is shown to be a reliable and accurate device to quantify the liveliness of an experimental rat. During recording the animal did not seem to be disturbed in any way by the device. It could stay in its normal cage, move freely, walk around and eat and drink ad libitum. Measurement did not require extra light, sound or other stimuli. Interpretation of the data was easy. The computer which samples the Doppler radar output signal generates activity curves which were easily interpreted for different ranges of vitality, varying between high liveliness and apnoea or cardiac arrest. The apparatus is low priced, and simple to build and use. PMID- 8231296 TI - Detection of electrocardiographic 'left ventricular strain' using neural nets. AB - The use of artificial neural networks for classification of ST-T abnormalities of the electrocardiogram (ECG) was investigated. A training set of 356 lateral leads selected from 105 ECGs was visually classified as exhibiting one particular ST-T morphology (left ventricular (LV) strain) or not. Selected measurements, together with the classification, were fed as input to a three-layer software-based network during the learning process. The performance of the network was evaluated by comparing the results obtained from the network with conventional criteria, using two test sets. Set 1 comprised 63 lateral leads from 32 ECGs with ST-T changes showing atypical forms of LV strain. Set 2 consisted of 80 lateral leads from 20 ECGs containing normal and abnormal T-waves. For set 1, the network outperformed conventional criteria, having a higher sensitivity (96 per cent against 85 per cent) and specificity (67 per cent against 50 per cent). With test set 2, both network and conventional criteria were 100 per cent sensitive and 100 per cent specific. For sets 1 and 2 combined, the network had a higher overall sensitivity (97 per cent against 89 per cent) and specificity (88 per cent against 82 per cent). The results suggest that neural networks may be useful in selected areas of electrocardiography, but care is required when selecting patterns for use in the training process. PMID- 8231297 TI - Biomechanical approach to quantifying anticipatory postural adjustments in the elderly. AB - The paper outlines a biomechanical approach to quantifying anticipatory postural adjustments in the elderly. The measurement problems that occur in applying the biomechanical approach to elderly subjects are described and the 'signal-to noise' properties of three candidate measures are compared, using data from volitional unilateral arm-raise tests performed on 100 elderly subjects. The results suggest that changes in vertical ground reaction force provide the greatest potential for accurate measurement of anticipatory adjustments, in comparison with changes in horizontal force or centre-of-pressure displacement. By normalising the anticipatory change in vertical ground reaction force with respect to the vertical perturbation force induced by the arm motion, a measure of relative anticipatory response is derived. The use of this measure, as well as its limitations, are demonstrated by analysing its relationship to actual falling risk, monitored prospectively in the elderly subject population. The findings showed evidence of larger relative anticipatory adjustments in the subjects who experienced recurrent falls, and it is suggested that these responses may be indicative of disordered motor programming. However, to detect these differences, it was necessary to average responses over multiple trials and to exclude trials with very small arm acceleration or very large baseline 'noise' (associated with ongoing postural sway). The need to screen and exclude data would seem to limit the practical utility of this approach in testing elderly populations. PMID- 8231298 TI - Wave propagation with different pressure signals: an experimental study on the latex tube. AB - To have deeper insight into the main factors affecting wave propagation in real hydraulic lines, we measured the true propagation coefficient in two latex rubber tubes via the three-point pressure method. The measurements were performed using both sinusoidal pressure signals of different amplitudes and periodic square waves as well as aperiodic pressure impulses. The results obtained were then compared with those predicted by a classic linear model valuable for a purely elastic maximally tethered tube. Our measurements demonstrate that the three point pressure method may introduce significant errors at low frequencies (below 1 Hz in the present experiments) when the distance between two consecutive transducers becomes much lower than the wavelength. The pattern of phase velocity in the range 2-20 Hz turns out to be about 10 per cent higher than the theoretical one computed using the static value of the Young modulus. This result supports the idea that the dynamic Young modulus of the material is slightly higher than that measured in static conditions. The experimental attenuation per wavelength is significantly higher than the theoretical one over most of the frequencies examined, and settles at a constant value as frequency increases. Introduction of wall viscoelasticity in the theoretical model can explain only a portion of the observed high frequency damping and wave attenuation. Finally, increasing the amplitude of pressure changes significantly affects the measured value of the propagation coefficient, especially at those frequencies for which direct and reflected waves sum together in a positive fashion. In these conditions we observed a moderate increase in phase velocity and a much more evident increase in attenuation per wavelength. PMID- 8231299 TI - Analysis of hand forces in health and disease during maximum isometric grasping of cylinders. AB - An analysis of force distribution in the hand during maximum isometric grasping actions is reported in a detailed and accurate manner. A microcomputer-controlled instrument which measures all 12 phalangeal forces of fingers simultaneously, in a single attempt at squeezing a cylindrical object, is described. The study involved 20 normal subjects of different weights and age groups grasping tubes of 50 mm, 75 mm, 90 mm and 110 mm diameters. Normal grasp forces decreased significantly with the increase in tube diameter, with the force being concentrated more on the distal segments of the fingers on the proximal and middle segments. The mean percentage contributions of finger forces to total grip strength, from index to little fingers, were 31, 33, 22 and 14 per cent, respectively. The study was extended to cover leprotic and paralytic hands to assess their functional capabilities. In the case of leprosy subjects, the grip strength decreased with the severity of the disease and was only about 50 per cent of that of normal subjects. In hemiplegics, the grip strength was only about one-eighth of the normal values. The above assessment procedure provides baseline data which could serve as guidelines to a clinician in assessing the severity of the disease and observing the patient's recovery following the treatment. It would also be useful in the design of hand-operated controls and prosthetic arms. PMID- 8231300 TI - Myocardial mechanics and the Fenn effect determined from a cardiac muscle crossbridge model. AB - A three-element cardiac muscle fibre model, utilising Huxley's sliding filament theory for the contractile element and coupled with parallel and series elastic components, was simulated to see if it were possible to predict the cardiac Fenn effect. The force/length energy (FLE) was computed in both isometric and isotonic contractions, as a function of muscle fibre length (preload) in the isometric case and afterload in the isotonic contraction case. Simulation results demonstrated that isotonic contractions produced a greater FLE than isometric contractions at every corresponding afterload, with the difference being equal to the work produced in the isotonic case, which is characteristic of the Fenn effect. The maximum energy utilisation was observed at maximum force isometric contractions, as has been experimentally observed in cardiac muscle. Changing the stiffness of the series elastic component did not change the Fenn-effect behaviour. Fenn-effect plots using crossbridge energy predictions showed behaviour similar to the FLE plots, but the FLE: crossbridge energy ratio declined with decreasing force even though the efficiency has been experimentally found to be constant. PMID- 8231301 TI - Microcomputer-based cardiac field simulation model. AB - Cardiac field simulation is one of the frontier subjects in electrocardiogram theory study. The paper describes a complete cardiac simulation model implemented on an IBM-PC/AT microcomputer. This model uses a new algorithm for excitation propagation simulation. In comparison with the previous rule-based algorithms, the new algorithm shows better in simulation speed and simulation accuracy. PMID- 8231302 TI - Simulation of the conduction velocity properties of nerve fibres by an electrical model. AB - The paper presents a simple electrical circuit model from which the relationship between the diameter of the axon and its conduction velocity can be derived. The model is primarily intended for the larger myelinated fibres in peripheral nerves. However, a simplified model is also shown for nonmyelinated fibres, where the relationship is quite different. In the conclusion some qualitative considerations are presented for thin myelinated fibres with a slow velocity of conduction. The conduction velocity/diameter relationship of these fibres gradually approaches that for nonmyelinated fibres with decreasing diameter. PMID- 8231303 TI - Kinematic analysis of human walking gait using digital image processing. AB - A system using digital image processing techniques for kinematic analysis of human gait has been developed. The system is cheap, easy to use, automated and provides useful detailed quantitative information to the medical profession. Passive markers comprising black annuli on white card are placed on the anatomical landmarks of the subject. Digital images at the standard television rate of 25 per second are acquired of the subject walking past a white background. The images are obtained, stored and processed using standard commercially available hardware, i.e. video camera, video recorder, digital framestore and an IBM PC. Using a single-threshold grey level, all the images are thresholded to produce binary images. An automatic routine then uses a set of pattern recognition algorithms to locate accurately and consistently the markers in each image. The positions of the markers are analysed to determine to which anatomical landmark they correspond, and thus a stick diagram for each image is obtained. There is also a facility where the positions of the markers may be entered manually and errors corrected. The results may be presented in a variety of ways: stick diagram animation, sagittal displacement graphs, flexion diagrams and gait parameters. PMID- 8231304 TI - Real-time measurement of the contribution of the muscular activity to the metabolic rate in freely moving rats. AB - A new real-time ergometric system, ERGORAT, for measuring the energy expenditure due to muscular activity of small mammals is described. The method is based on measuring the vibrations induced by a freely moving rat to a platform on which it is living. Six accelerometers placed on the platform detect all the vibrations produced by the rat. The co-ordinates of the centre of mass of the animal are located by an optoelectronic device. This location and the six accelerations are fed to a microprocessor-based data acquisition and processing system. Using a Lagrangian dynamic model, the values of the mechanical energy transferred to the platform are computed every second. ERGORAT is used as part of an experimental setup allowing the measurement of the energetic balance of lean and obese rats. The results clearly show that the increase in metabolic rate of obese rats during cold exposure can be entirely explained by the cost of their increase in locomotor activity, whereas for lean rats, this cost accounts for only 41.5 per cent of their metabolic increase, the remaining being the contribution of their active brown adipose tissue. PMID- 8231305 TI - Detection of fetal breathing and cardiac movements and rhythms in ultrasonic Doppler signal recorded on the surface of the maternal abdomen. AB - It is proposed to use information on the direction of reflector movement and extensive filtering in the detection of fetal breathing and cardiac movements in the ultrasonic Doppler signal recorded on the surface of the material abdomen. The method appears fairly insensitive to spurious signals and allows those of interest to be distinguished without any reference technique. A decision rule for breathing and cardiac rhythm detection, incorporating movement direction, amplitude, shape and periodicity criteria, is also proposed. PMID- 8231306 TI - Continuous measurement of blood volume using light reflection: method and validation. PMID- 8231307 TI - Digital filters for the detection of late potentials in high-resolution ECG. PMID- 8231308 TI - Conscious mental tasks and their EEG signals. PMID- 8231309 TI - Cerebral blood flow measurement using the hydrogen clearance technique: a microcomputer-based system for data collection and analysis. PMID- 8231310 TI - Transducer for the measurement of linear displacement of body segments. PMID- 8231311 TI - Quality of electrical mains supplies in hospitals: implications for microprocessor-controlled medical equipment. PMID- 8231312 TI - Rate-responsive pacing based on sympathetic activity. AB - Modern pacemaker technology makes the adaptation of pacing rate to haemodynamic requirements possible. The most ambitious approach aims at restoration of the physiological closed-loop system by utilising the information that is supplied by the autonomic nervous system (ANS) and extracted from myocardial contractile performance. Measurement is accomplished by the impedance method, with the stimulation electrode as measuring electrode. The ventricular inotropic parameter (VIP) has been identified as an ANS-dependent parameter. A special detection algorithm RQ (regional effective slope quantity) with high ANS sensitivity has been developed. Rate adaptation has been achieved by using an individually adjustable inotropic index (II). The concept has been evaluated in a multicentre study employing a standardised exercise protocol. The results in patients with AV block demonstrate the excellent agreement between the spontaneous sinus rhythm and the ANS-controlled stimulation rate during different forms of exercise. Measurement of mean arterial blood pressure (MABP) supports the physiological approach to adapting the pacing rate to different kinds of haemodynamic challenge. PMID- 8231313 TI - Rhythmic oscillations of blood pressure and R-R interval in patients with chronic renal failure. AB - Rhythmic blood pressure (BP) and R-R interval (R-R) oscillations at low-, mid- and high-frequency bands (LF: 0.02-0.06 Hz; MF: 0.07-0.14 Hz; HF: 0.15-0.40 Hz) were compared between uraemic patients maintained on haemodialysis and control subjects. The LF and MF power spectra of BP were attenuated more in patients than in controls. With subjects standing, the MF power spectrum of BP increased significantly in both groups. With subjects supine, the plasma norepinephrine concentration was higher, and its increment upon standing was greater in patients than in controls. Each R-R frequency power spectrum decreased more in patients than in control subjects. The HF power spectrum of R-R, i.e. a vagal tone index, systematically decreased upon standing in the control subjects but not in the patients. The linear coupling between BP and R-R oscillations was strongest in the HF band, decreasing in the MF and LF bands. Transfer function analysis indicated that, in uraemic patients, linear BP/R-R relationships were altered in the HF band but remained normal in the LF and MF bands. The present results suggest that, first, the decreased amplitude of Mayer waves, i.e. the MF power spectrum of BP, observed in uraemic patients can be attributed to low sensitivity of the vasculature to sympathetic stimuli, and, secondly, autonomic modulation of linear BP/R-R relationships is frequency-dependent. PMID- 8231314 TI - Ariel: an information system for AIDS research. AB - An information system has been established at the National Institute for Biological Standards & Control for the exchange of knowledge in AIDS research, particularly in relation to vaccine design. This system, the AIDS information exchange link (Ariel), is designed to act as a central store of relevant information for scientists in the UK and abroad and was set up under the auspices of the Medical Research Council. It holds information on research materials (reagents), on genetic sequences and on projects. Several computers and database systems are involved. Access is obtained through Janet, the UK academic network, or PSS, the British Telecom public network. Both these networks are linked to international network systems, e.g. Internet, Earn, IPSS. Ariel has been in operation for 18 months and is accessed internationally. PMID- 8231315 TI - Haemocompatibility optimisation of implants by hybrid structuring. AB - State of the art in biomaterial research and implant design is a compromise between functionality and biocompatibility. Consequently, results often have disadvantages with respect to both aspects. With regard to biocompatibility, the activation of the clotting system by alloplastic materials is of great significance, because it necessitates anticoagulant therapy. Further improvements in implant technology require an understanding of the interactions between blood and implants. Therefore a microscopic model of thrombogenesis at alloplastic surfaces is briefly presented, relating thrombogenicity of a material to the electronic structure of its surface. The electronic requirements for high haemocompatibility, which result from this model (especially a low band-gap density of states and a high surface conductivity) are fulfilled by an amorphous alloy of silicon and carbon (a-SiC:H). The advantage of amorphous materials is that they do not obey stoichiometric rules. Thus they allow a continuous adjustment of the electronic parameters without fundamental changes in their mechanical and chemical properties. The theoretical results were checked in vitro by total internal reflection intrinsic fluorescence (TIRIF) spectroscopy as well as thrombelastography experiments (TEG). In comparison with conventional materials such as titanium or LTI carbon, the TEG-clotting time of a-SiC:H coatings was prolonged by in excess of 200 per cent. As a consequence, a-SiC:H is well suited as a haemocompatible coating material for hybrid structuring of cardiovascular implants. PMID- 8231316 TI - Asymmetrical loads and lateral bending of the human spine. AB - The human spine is modelled as a cantilever-type beam column. Under the influence of static asymmetrical loads, muscle and low-back forces are predicted from a hypothetical but revealing model. Such forces produced by asymmetrical loads are much larger than for a corresponding symmetrical load. Asymmetrical loads can encourage, especially in young schoolchildren, lateral bending of the spine by alleviating muscle and low-back forces. This could possibly be a factor contributing to the surprisingly high percentage of schoolchildren with measurable scoliotic curves. The wearing of knapsack-type bags is advocated. PMID- 8231317 TI - Fast and reliable QRS alignment technique for high-frequency analysis of signal averaged ECG. AB - The process of QRS alignment as required in signal-averaged ECG can impose serious limitations on the spectral range of the signal output. This effect depends basically on the particular alignment technique being used and on the level and type of noise present in the recorded ECG. In clinical studies where a wide-band (1000 Hz) ECG averager is required, the conventional QRS alignment technique, based on maximum coherence matching (MCM) with a template beat, may not perform consistently well. An alternative QRS alignment technique based on the accurate detection of a single fiducial point (SFP) in the bandpass filtered (3-30 Hz) QRS complex was developed. Using computer simulation methods, a comparative assessment of the frequency bandwidths (3 dB points) offered by both MCM and SFP techniques as a function of noise level (15-100 muRMS) and type (EMG and 50 Hz interference), was carried out. The results of the comparative assessment indicated a better performance by the SFP technique in all cases of noise. Hence, the SFP technique would perform more reliably for high-frequency analysis of a noisy ECG, especially when 50 Hz interference is high. Furthermore, SFP is considerably faster than MCM (about four times) when implemented digitally, and its analogue realisation is feasible. The SFP technique is suitable for late-potential analysis in the signal-averaged ECG. PMID- 8231318 TI - Biodegradable copolymers based on p-dioxanone for medical application. PMID- 8231319 TI - Estimation of organ biodistribution and absorbed dose from external measurement with TLDs in PET studies. PMID- 8231320 TI - Noninvasive measurement of blood glucose concentrations by analysing Fourier transform infra-red absorbance spectra through oral mucosa. AB - Whether Fourier transform infra-red spectroscopy with an attenuated total reflection prism could be applied for noninvasive glucose measurement through oral mucosa was evaluated. As a result, the same absorbance peak at 1033 cm-1 as in glucose aqueous solution was found in the absorbance spectra through mucous membrane. However, these glucose specific peaks were interfered with by the baseline drifts owing to prism attachment and the background spectra from body constituents other than glucose. Therefore, to eliminate these interferences, the calibration curve between the second derivatives of the absorbance peak at 1033 cm-1 and those at 2920 cm-1 was calculated (r = 0.910). By using this calibration curve, the spectral changes due to prism attachment were first eliminated. Secondly, by obtaining the second derivative of the difference between the postprandial absorbance peak and the fasting sample as a characteristic of an individual, high correlations between the corrected second derivatives of absorbance spectra through the mucous membrane of the lip at 1033 cm-1 and the increases in blood glucose concentrations above fasting levels were observed (r = 0.910). In conclusion, it was suggested that Fourier transform infra-red spectroscopy could be useful for noninvasive monitoring of glucose through oral mucosa. PMID- 8231321 TI - Present and future status of noninvasive selective deep heating using RF in hyperthermia. AB - To achieve hyperthermia using electromagnetic energy, RF of under 100 MHz is basically suitable for the external heating of the deep portions of the body. For applicators using such RF, the following types are considered: capacitive, inductive, radiative and hybrid. With radiative applicators, the intensity of the EM waves radiated from the applicator decreases with propagation into the material to be heated, but the phased annular array of radiative applicators potentially increases the intensity of the EM energy in the deep portion owing to the interference of the waves. Using this method, the focusing of EM energy depends on the dielectric properties of the material to be heated. With respect to RF heating at a lower frequency than the RF used for the annular phased array, some devices have been said to concentrate EM energy in the deep portions, where the characteristics of 'wave' are not utilised. To this end, some methods using capacitive electrodes, an inductive coil, or a combination of both, are being designed. The results of using such methods have shown that it is possible to supply sufficient EM energy to the muscle layers deep in the material to be heated, without heating the fat layers excessively. PMID- 8231322 TI - Optimisation of dose distribution for linear accelerator-based stereotactic radiosurgery. AB - The work presented in the paper addresses a method for obtaining the optimal dose distribution for LINAC-based stereotactic radiosurgery. As many targets have nonspherical or irregular shapes and three-dimensional dose calculations included in dose optimisation, long computation times are required to determine the optimum isocentre separation and collimator sizes to shape the irregular target using the multiple-isocentre approach, by trial-and-error types of method. The simple approach, using a computer-aided design optimisation technique and a fast approximate dose model, has been developed to find the optimum isocentre positions and collimator sizes quickly and automatically. A spherical dose model has been developed to represent the dose for a standard arc system with a single isocentre. The implementation of computer-aided design algorithms with the spherical dose model and their application to several cases are discussed. It is shown that the spherical dose model gives dose distribution similar to that of the exact dose model, which makes this simple dose model more efficient, with computer-aided design optimisation, in finding optimum isocentre positions and collimator sizes used in stereotactic radiosurgery. PMID- 8231323 TI - Tomographic measurement of temperature change in phantoms of the human body by chirp radar-type microwave computed tomography. AB - The chirp radar-type microwave computed tomograph (CT) measures the temperature change in a human body noninvasively. The paper examines its feasibility. A chirp pulse signal between 1 and 2 GHz is radiated from the transmitting antenna to the phantom. The transmitted waves are detected by the receiving antenna, which is placed on the opposite side of the object, and the beat signal between the incident wave and the transmitted wave is produced by the mixer. By spectral analysis of the beat signal, only those signals transmitted on the straight line between the transmitting antenna and the receiving antenna are discriminated from multipath signals. The microwave tomogram can therefore be reconstructed easily using the conventional algorithms for an X-ray CT image. The microwave CT can use the chirp signal to remove the influence of multipath signals caused by diffraction and reflection. The imaging of dielectric materials with complicated structures is thus possible. The experimental results using phantoms show that the spatial resolution of this microwave CT is about 10 mm and that a two dimensional distribution of temperature change can be measured. PMID- 8231324 TI - Disk-cathode flash X-ray tube driven by a repetitive two-stage Marx pulser. AB - Fundamental studies of a repetitive flash X-ray generator using a disk-cathode radiation tube are described. The high-voltage pulser employed a modified two stage surge-Marx circuit. The two condensers in the pulser were charged from 40 to 60 kV, and the electric charges were discharged to the X-ray tube repetitively to generate flash X-rays. The total capacity during the main discharge was 425 pF, and the maximum output voltage from the pulser was about 1.9 times the charged voltage. The flash X-ray tube was of the demountable-diode type and was composed of a rod-shaped anode tip made of tungsten, a disk cathode made of graphite and a tube body made of polymethylmethacrylate. The peak tube voltage was primarily determined by the anode-cathode (A-C) space, and the peak tube current was less than 0.5 kA. Thus the maximum photon energy could be easily controlled by varying the A-C space, and the tube current roughly increased according to increases in the charged voltage. The pulse width ranged from 40 to 100 ns, and the X-ray intensity was less than 1.1 microC kg-1 at 0.5 m per pulse. The repetition rate was less than 50 Hz, and the effective focal spot size was equivalent to the anode diameter. PMID- 8231325 TI - Characteristics of proton beams after field shaping at PMRC. AB - The proton irradiation control system was developed for cancer radiotherapy at the Proton Medical Research Center, with the extension of a beam line connected to a synchrotron at the High Energy Physics Laboratory. The initial energy of the 500 MeV proton beam supplied by the accelerator is degraded down to 243 MeV after passing through a graphite rod. In the control system, a proton beam is scattered to form a large field, its Bragg peak width is spread out, and its energy is degraded to the optimum value with a range covering tumour depth. The characteristics of the devices required for these procedures have been investigated from the viewpoint of the relationship between dose rate and field flatness, taking the setting-up geometry of these devices into consideration. PMID- 8231326 TI - Haemodynamic changes in IUGR fetus with chronic hypoxia evaluated by fetal heart rate monitoring and Doppler measurement of blood flow velocity. AB - The measurement of fetal blood flow velocity and 24 h monitoring of fetal heart rate (FHR) using a computer were performed to clarify the haemodynamics of growth retarded fetuses with chronic hypoxia. One hundred normal-growth and 18 growth retarded fetuses were analysed. All the growth-retarded fetuses with chronic hypoxia were characterised by abnormal blood flow velocity waveforms (with the pulsatility index in the descending aorta below the -1.0 SD and in the middle cerebral artery above the +1.0 SD for our reference range, from 100 normal-growth fetuses). In the latter, the incidence of accelerations of defined size and variability in FHR patterns showed a diurnal variation after 30 weeks' gestation. The initial change in FHR patterns during hypoxia in 11 growth-retarded fetuses, resulting in fetal distress, was a derangement of diurnal variations in FHR patterns, followed by a decrease in variability. A rapid increase in blood flow velocity in the middle cerebral artery with the advance of hypoxia was observed before the onset of distress. Maternal low-dose oxygen inhalation elicited a temporary increase in FHR variability in the growth-retarded but not in normal fetuses. Re-inhalation after 1 h elicited a similar change, suggesting that intermittent rather than continuous, oxygen inhalation may be more effective. PMID- 8231327 TI - Measurement of fetal movements using multichannel ultrasound pulsed Doppler: autorecognition of fetal movements by maximum entropy method. AB - Changes in fetal movements indicate biophysical conditions and functional development. The precise evaluation of fetal movements in clinical medicine requires the development of a continuous automated monitoring technique. A basic study of the measurement of fetal movements was carried out by modifying the Doppler ultrasound module of a cardiotocograph to produce low-frequency Doppler signals and five simultaneous outputs at various depths. These outputs represent displacement inside tissue at the various depths. Signal processing was executed on a 32-bit computer with a high-accuracy displacement estimation technique using the arctangent method. Results showed successful tracking of minute movements, such as fetal breathing movements (FBM), while rejecting other movements derived from maternal breathing etc. Using spectral analysis by the maximum entropy method (MEM), fetal movements were classified in three groups (FBM, fetal gross movements (FGM) and fetal heart movements (FHM), based on the character of their special peak frequencies. The order of movement recognition was first FGM, then FBM and lastly FHM. FBM were more successfully recognised by MEM than by conventional B-mode observation methods. Small body movements were difficult to recognise as FGM by MEM in some cases. Although further studies are required for clinical application, it appears that automated assessments of fetal movements should be possible with this technique. PMID- 8231328 TI - Usefulness of sodium chloride as a nondiffusible indicator in the measurement of extravascular lung thermal volume in dogs. AB - The authors examined the usefulness of sodium chloride as a nondiffusible indicator during the first passage through dogs' lungs, before and after increased-permeability pulmonary oedema produced by an intravenous injection of alloxan. With an injection of a mixture of ice-cold 3 per cent sodium chloride and indocyanine green dye (a nondiffusible reference indicator), the authors simultaneously recorded three dilution curves from the aortic root: dye dilution, thermal and blood electrical conductivity dilution curves in six dogs. The mean transit time of sodium chloride in the conductivity dilution curve was significantly different from, but fairly equal to, that of indocyanine green dye (6.2 +/- 1.4 s (mean +/- SD) against 6.5 +/- 1.4 s (p < 0.01) in the baseline period, and 7.6 +/- 1.9 s against 8.4 +/- 2.1 s (p < 0.01) in the oedema period, respectively). The calculated extravascular lung thermal volume with the thermal and conductivity dilution method (Y, ml kg-1) correlated well with the gravimetrically determined extravascular lung mass in a total of 12 dogs, including six other dogs without intervention (x, g kg-1) (y = 0.72 x +3.03, r = 0.96). The authors conclude that sodium chloride is useful as a nondiffusible indicator in the first passage through the lungs, and that the thermal and conductivity dilution method is also useful for measuring extravascular lung water mass. PMID- 8231329 TI - Measurement of the micro-electroretinogram and component analysis. AB - The authors propose a new technique for detecting micro-electroretinograms (muERG) which uses light-emitting diode (LED) stimulation and frequency analysis. The advantage of this method is the applicability of linear system analysis to a muERG obtained by partial stimulation of a computer-controlled waveform. The paper discusses two basic techniques for detecting the muERG. One is Fourier analysis of a synchronised ERG obtained by sine-wave stimulus, and the other is correlation analysis using a random stimulus. The results of the two methods are compared, and the usefulness of the proposed method is indicated; the muERG is analysed by means of a model consisting of five components of a second-order transfer function with delay. Functions of the central and peripheral areas of the retina could be analysed in more detail by this technique. PMID- 8231330 TI - Matching between theoretical and experimental data for ELF ion transport effects. AB - In recent years, several studies have been focused on the problem of nonthermal interaction between extremely low-frequency (ELF) electromagnetic fields and cell environment at membrane level. In the paper, to analyse the dynamic effects of weak static and harmonic fields on charged particles, some new considerations have been developed, based on the Lorentz model. The authors have reached a suitable formulation so that the data processing has led expressly to the evaluation of ionic-velocity components against magnetic-field amplitudes and frequencies, as well as a viscosity parameter. Even through a direct and rough comparison, the results of the authors' investigation have demonstrated an interesting agreement with some experimental data relative to ionic fluxes through cell membranes. Indeed, by means of an algorithm based on the techniques of the inversion theory, the author's have found definite values of the viscosity parameter for which the expected resonant behaviours (amplitude and frequency windows) fit existing experiments well. It seems worthy of interest that such viscosity values fall within a consistent, narrow range of low amplitude. PMID- 8231331 TI - Cable equation for a myelinated axon derived from its microstructure. AB - A simplified cable equation that describes the subthreshold behaviour of a myelinated axon is derived from its microstructure. Specifically, a microcontinuum cable model of a composite axon is homogenised, yielding a familiar macrocontinuum cable equation of electrotonus, for which the space and time constants depend on microstructural electrical parameters. Activating functions for magnetic and electrical stimulation can be incorporated into this homogenised cable equation as sources or sinks of transmembrane potential. An integral solution to the forced cable equation is also presented for the subthreshold regime. Errors are introduced when myelin membrane resistance is assumed to be infinite. PMID- 8231332 TI - Application of multivariate autoregressive modelling for analysing chloride/potassium/bicarbonate relationship in the body. AB - The authors repeatedly analysed course data of acid-base disturbances accompanying hypochloraemia and/or hypokalaemia by means of multivariate autoregressive modelling. It was found that the regulatory relationship between chloride and bicarbonate is inverse between the following two hypochloraemic hyperbicarbonataemic states: the one induced by chloride depletion and the other induced by CO2 retention. Also, the study revealed an independent role of potassium deficiency in the development of metabolic alkalosis, especially in cases with mineralocorticoid-induced alkalosis. The present approach enabled the authors to solve a long-standing problem, i.e. to differentiate between the roles of chloride and potassium in the development of metabolic alkalosis. PMID- 8231333 TI - Do patients' evaluations of a future health state change when they actually enter that state? AB - Fundamental to using utilities for future health states for clinical decision analysis is the assumption that the utilities are stable. The purpose of this study was to determine if cancer patients' prior evaluations of a treatment induced health state remain stable when they later experience that state. Before a four-week course of radiation therapy, 66 laryngeal cancer patients evaluated three common treatment-induced outcome scenarios (mild/moderate/severe) using rating scales and the time trade-off, and provided self-assessments of voice symptoms, voice function, and general health. Evaluations and self-assessments were repeated at the end of therapy, and respondents indicated which outcome scenario described their actual end-of-therapy state. Twenty-four patients identified the mild scenario as their outcome state, 36 the moderate, and 6 the severe. No across-time differences in evaluations were detected, although significant (P < .0001) downward shifts in scores for voice symptoms (t = 5.67), voice function (t = 5.10), and general health (t = 6.69) indicated that actual change in health status had occurred. These results imply that patients' evaluations of imagined short-term health states encountered during radiation therapy for laryngeal cancer remain consistent when those states are experienced at a later time. The design of this project provides a model for determining if this consistency appears in other clinical situations. PMID- 8231334 TI - Health insurance and utilization of medical care for children with special health care needs. AB - Based on analyses of the 1988 National Center for Health Statistics, National Health Interview Survey, Child Health Supplement, this study examines the insurance coverage and utilization of physicians, hospitals, and prescribed medicine among the 9.6 million U.S. children with special health care needs, defined as children who had one or more selected chronic conditions that caused them to experience pain, discomfort, or being upset often or all of the time in the last year, or who were limited in their major childhood activities (playing or going to school) as a result of these or other impairments or health problems. The findings confirmed that substantial variation in access to routine medical care exists among these children. Though health status is an important predictor of which children use services in general, poor, minority children who lived with their mother or someone other than their parents, or those without insurance or an identifiable regular medical provider were most likely to experience financial barriers to access or were less apt to seek care than other children with comparable needs. Access to routine medical care remains particularly problematic for these subsets of children with special health care needs. PMID- 8231335 TI - Access to heart and liver transplantation in the late 1980s. AB - Because of a shortage of usable organs, many who require heart or liver transplants for survival will not have access to them. Access to care may reflect demographic factors and ability to pay, as well as medical considerations. Receipt of an organ may be influenced by expected survival with and without a transplant, age, gender, race, ability to pay, and distance to a transplant center. Discharge abstract data from a national sample of over 500 hospitals in 1986 and 1987 were used to select heart and liver recipients and others with end stage diseases who did not receive a transplant. Multivariate logistic regression analyses were then used to estimate how receipt of a transplant was influenced by expected years of survival after transplantation (YAT), expected ability to pay, age, sex, race, and distance to a transplant center. Controlling for differences in expected YAT, age, sex, race, and distance to the transplant center, those expected to have the most ability to pay were more likely to receive heart and liver transplants, compared to those expected to have medium ability to pay. Third-party coverage was particularly important in receipt of a transplant for those with absolute contraindications. Expected YAT and age were significant, with some evidence of a tradeoff between urgency and expected YAT in the case of hearts. Men were more likely to obtain heart transplants and women were more likely to get liver transplants. The effects of distance were small. Existing regulatory incentives and biological, medical, and cultural reasons may justify the age-, sex-, race-, and prognosis-related differences in the odds of receiving a transplant. The importance of ability to pay may not have been adequately observed in previous studies restricted to the patients screened at major transplant centers. Hospital discharge records with personal identifiers, linkage to official waiting lists, and better patient level socioeconomic information would permit more definitive analysis. PMID- 8231336 TI - Rating journals in health care administration by the textbook citation method. PMID- 8231337 TI - Nursing home ownership revisited: market, cost and quality relationships. PMID- 8231338 TI - Hospital corporate restructuring and financial performance. AB - In the last decade, an important innovation in the organizational structure of acute care hospitals occurred. Many hospitals restructured by creating subsidiaries that segment assets or services into separate corporations. We know relatively little about the effects of such restructuring. This paper examines the association of restructuring with financial performance of not-for-profit hospital firms. The study uses data from all not-for-profit acute care hospital firms in Virginia, the only state for which the unique study data are available. We find that the consolidated financial performance of hospital firms is influenced by factors that affect the hospital's financial performance (i.e., payer-mix, staffing and service mix) but not the number or size of non-hospital subsidiaries. Future research should examine the effect of restructuring on other types of performance. PMID- 8231339 TI - Measuring physicians' tolerance for ambiguity and its relationship to their reported practices regarding genetic testing. AB - Despite uncertainties in medicine, attempts to study physicians' tolerance for uncertainty have been few, and limited by the measurement instruments available. This paper describes development of a modified tolerance for ambiguity (TFA) scale, and correlates it with several physician characteristics and reported behaviors. Eighteen TFA items were included in a national survey of physicians' knowledge and attitudes about genetic testing. Sixty-five percent (n = 1,140) of 1,759 obstetricians, pediatricians, internists, family practitioners, and psychiatrists responded. After psychometric analyses, the scale was reduced to 7 items, demonstrating an acceptable reliability (Cronbach's alpha = .75). TFA was higher among psychiatrists than other specialties, among those who were older when they graduated from medical school, and among those willing to offer a new low-cost, accurate predictive test when none of their colleagues do. TFA was lower among those who indicated that attendance at religious services was important, among those who would make a recommendation to their patients regarding pregnancy termination after prenatal diagnosis, and among those who would withhold negative genetic test results. Future research is needed on the scale itself, and to assess factors affecting TFA, such as its susceptibility to modification, and its potential association with clinical practice in other areas of medicine that are characterized by ambiguity. PMID- 8231340 TI - [Effectiveness of the treatment of systemic lupus erythematosus with methotrexate]. AB - BACKGROUND: To evaluate the efficacy of methotrexate in patients with systemic lupus erythematosus (SLE) without major organ involvement resistant to medium high doses of prednisone. METHODS: Crossover, open clinical trial with two treatment periods, the first of 3 months and the second of 6 months, an intermediate control period of 3 months and another at the end of 6 months. A sample of 15 consecutive patients with SLE who, with no major organ damage, had active disease in spite of receiving more than 10 mg/day of prednisone or who relapsed on reduction of this doses during a period of at least 3 months. 7.5 mg/week of methotrexate were administered orally, divided into three doses of 2.5 mg/12 hours. Statistical significance was evaluated by Student's paired t test and chi 2; the strength of association by the Mantel-Haenzel odds ratio (OR) method and the precision, by Miettinen's confidence interval (CI). A p value of less than 0.05 was considered significant. RESULTS: Two patients failed to finish the study; one for worsening of cutaneous lesions of necrotizing vasculitis which she already had previously, and the other for an increase in her transaminase levels. In the remaining 13 there were 10 flares of disease activity during the control phases, 2 severe, versus 2 flares during the periods of methotrexate use (OR 7.69 (95% confidence interval, 1.67 to 33.33; p = 0.021). There were no significant changes in analytical results or prednisone requirements. During treatment six patients had oral aphthae and five had dyspepsia; three had an increase in transaminase levels, which in one caused the treatment to be stopped. There were two urinary infections, one community acquired pneumonia and one upper airway symptoms requiring antibiotic treatment; one female patient had acute cholecystitis with cholelithiasis necessitating surgical intervention. CONCLUSIONS: Weekly low doses of methotrexate may prevent flares of activity of SLE in this type of patients, but it does not reduce the requirements of prednisone, nor modify analytical data. Toxic effects are rare and reversible upon interrupting medication. PMID- 8231341 TI - [Reactivity of serologic tests for the detection of syphilis in patients infected with the human immunodeficiency virus]. AB - BACKGROUND: The coexistence of syphilis and infection by the human immunodeficiency virus (HIV) appears to modify the natural history of both diseases. The aim of this study was to know the prevalence of syphilis in a population of patients with HIV infection, the possible association with certain risk practices and the validity of the reaginic test in such patients. METHODS: Three hundred sixty-seven patients with HIV infection who went for the first time to a monographic clinic of a university hospital were studied. Syphilis serology was carried out: rapid plasma reaginic (RPR) and hemagglutination (MHA-TP) tests. RESULTS: Out of all the patients 26 (7.1%) had positive MHA-TP. The proportion of homosexuals was greater among those who had a positive treponemic test (69%) than among those who were negative (6.4%; odds ratio [OR] = 32.6; confidence interval 95%: 16.2-65.4). The positivity of MHA-TP was more frequent among those presenting criteria of the acquired immunodeficiency syndrome (AIDS) at the diagnosis (18% versus 5.6%; OR = 3.6 [1,5-8,9]). Seventy-four false positive reactions were observed with the RPR (20%) corresponding almost exclusively (96%) to intravenous drug users who presented false positivity in 25% of the cases. CONCLUSIONS: The prevalence of syphilis detected by treponemic serology among subjects with infection by the human immunodeficiency virus is related with homosexuality as the principal practice of risk. One quarter of the intravenous drug users with infection by the human immunodeficiency virus presented false positive results to the reaginic test thus leading to the recommendation that therapeutic measures should not be initiated without confirmation with a treponemic test. PMID- 8231342 TI - [Spanish scientific production on drug dependence]. AB - BACKGROUND: The investigation of drug dependency has been strengthened in the last few years in Spain. One indicator of the scientific production in one area is constituted by the analysis of the studies included in data bases. METHODS: The Spanish publications in the field of drug dependency from 1985-1990 were reviewed. The references of studies published by Spanish authors collected in MEDLINE with Medical Subject Headings (MeSH) corresponding to Substance abuse or Substance dependence were selected. The following studies were collected a) those in which Spain was reported as the country of the author; b) published in Spanish journals; c) those in which Spain was indicated as the MeSH; b) and c) were not included if the publications did not correspond to Spanish authors. RESULTS: Three hundred thirty-one references were collected; 273 corresponded to Substance dependence and 58 to Substance abuse. The two themes most frequently studied were dependent on heroin (n = 99) and alcoholism (n = 94). Most studies were done in universities, mainly in schools of Medicine. Forty-three percent of the papers were signed by three or four authors. There were 1,044 authors; 844 (81%) authors published one article and 9 published seven or more. Twenty-three percent of the articles were published in 1988 and 20% in 1989. Three hundred thirty-one studies were published in 78 journals. Sixty percent of the journals published one article and 17% two. The journal which published the most articles were Medicina Clinica (n = 112), and Revista Clinica Espanola (n = 40). The foreign journal which published the most articles was Drug and Alcohol Dependence (n = 11). Twenty-nine articles were published in 7 of the journals with greatest impact factor in this area. CONCLUSIONS: These results confirm the tendency observed in other studies and indicate the increase in the quantity and quality of the Spanish publications on drug dependence. PMID- 8231343 TI - [Do new treatments contribute to improve the prognosis in systemic lupus erythematosus?]. PMID- 8231344 TI - [The decree on selective drug financing in charge of the Social Security]. PMID- 8231345 TI - [Mitral endocarditis caused by Staphylococcus aureus resistant to methicillin, aminoglucosides and rifampicin: description of 2 cases with fatal course]. AB - Two patients with infectious endocarditis (IE) by Staphylococcus aureus resistant to methicillin, aminoglucosides and rifampicin (SARMAR) acquired in hospital during the course of an epidemic outbreak of this microorganism in the Hospital Clinic i Provincial of Barcelona. Both patients had undergone surgery of the lower limbs. The entrance of the microorganism was the infection of the surgical wound, with bacteriemia, followed by mitral IE after a short time interval (20 days). Despite adequate treatment with vancomycin both patients died. The culture of mitral vegetation was positive for SARMAR in one. Analysis of the chromosomic DNA of all the isolations from the patients was identical and coincided with that of the SARMAR strains isolated in the epidemic outbreak of the hospital. The current situation of IE by SARMAR is reviewed and the therapeutic implications commented upon suggesting that treatment of this entity should simultaneously include the administration of vancomycin and phosphomycin or cotrimoxazole, with surgery being considered if infection persists. PMID- 8231346 TI - [Diagnosis of ornithine carbamoyl transferase deficiency and heterozygote detection with allopurinol loading test]. AB - BACKGROUND: Allopurinol loading test is based on the inhibition of pyrimidine biosynthesis and the subsequent increase in orotic acid excretion caused by a single dose of allopurinol. Abnormally elevated amounts of orotic acid excretion are demonstrated in ornithine carbamoyl transferase (OCT) deficiency patients and heterozygotes as well as in other disorders of urea cycle. Biochemical studies performed for the diagnosis of one patient and carrier detection in her family are presented. METHODS: Amino acids: ion exchange chromatography; ammonium: method of Van Anken and Shiphorst; orotic acid: modification of Adachi et al, and allopurinol test following Brusilow et al. RESULTS: The characteristic amino acid profile of the patient together with her clinical history suggested the diagnosis of OCT deficiency, which was confirmed with protein and allopurinol loading test. The heterozygote condition became evident only by means of allopurinol test in 2/5 female relatives. CONCLUSIONS: Allopurinol test is a useful tool for the preliminary investigation of urea cycle function, avoiding the possible hyperammonemia caused by other test, and permitting extensive familial studies without hospitalization. It results more informative than the protein loading test. PMID- 8231347 TI - [Pedagogic orientation and medical libraries]. PMID- 8231348 TI - [Critical review of the indications of diagnostic endoscopic retrograde cholangiopancreatography in the 90s (and II)]. PMID- 8231349 TI - [Sudden death and sports]. PMID- 8231350 TI - [Form quality of mortality statistics in Salamanca and province]. PMID- 8231351 TI - [Cerebral infarction as the first manifestation of polycythemia vera]. PMID- 8231352 TI - [Prognostic value of serum neurospecific enolase in small cell bronchial carcinoma]. PMID- 8231353 TI - [A case-control study of risk factors of acute cerebrovascular disease]. AB - BACKGROUND: Stroke is the cardiovascular disease which causes the greatest number of deaths in Galicia, mortality, particularly in women, being higher than in the rest of Spain. The aim of this study was to investigate the risk factors of stroke and its importance in Galicia. METHODS: A hospitalary study of cases and controls was performed including 76 patients with stroke and 76 controls individually paired for age, sex, population habitat and date of admission with anamnesis, weight, height and blood pressure, analysis and electrocardiogram being carried out in all. RESULTS: An association was found between the disease and family history of stroke (odds ratio = 3.6, confidence interval 95% = 1.2 13.3), personal history of stroke (17.9; 4.0-79.1), personal history of atrial fibrillation (15.0; 3.3-68.3), high blood pressure (4.5; 1.9-11.6) and ingestion of alcohol greater than or equal to 80 g/day in comparison with abstemious patients adjusted for the effect of high blood pressure (2.5; 1.1-5.7). An association was also observed with serum cholesterol levels greater than or equal to 250 mg/dl (6.46 mmol/l) (3.3; 1.2-8.8, in comparison with cholesterol less than 200 mg/dl). No association was found with the cigarette smoking (1.2; 0.7 2.3). CONCLUSIONS: The results observed for high blood pressure and the ingestion of alcohol regardless of the same are of importance in primary prevention due to being modifiable risk factors. The association with family history of stroke and auricular fibrillation reinforce the needs for primary prevention measures in these subpopulations while having had a stroke or a transitory ischemic attack is the characteristic with the most risk reinforcing the need for secondary prevention measures which have found to be effective. The controls presented abnormally low cholesterol levels that do not reflect those of the population from which they originate, thus existing the possibility that the association observed is spurious. PMID- 8231354 TI - [Follicular mantle lymphoma: clinicopathologic and cell proliferation of 11 cases]. AB - BACKGROUND: The aim of the present was to study the biologic behavior of the lymphomas of the follicular mantle by analysis of different indexes of neoplastic cell proliferation and their relation with the clinical symptoms, prognosis and survival of the patients. METHODS: Light microscope, ultrastructural, and frozen and paraffin immunohistochemical studies were performed including the proliferative markers Ki-67 and PC10, flow cytometry and analysis of cell cycle in biopsy samples. Clinical data of 11 patients were collected in addition to therapeutic response and survival. RESULTS: Lymphoma of the follicular mantle is constituted by small sized cell elements with a variable pattern of tumoral growth in the lymph nodes. In normal lymphocytes of the follicular mantle the immunophenotype shows expression of IgM + IgD and a light chain in the cell surface. From a clinical point of view, massive splenomegaly and disseminated stage are the most outstanding characteristics. One third of the cases analyzed presented an aneuploid neoplastic population. According to the PC10 marker, the proliferative index ranged from 2.9 to 14.7% of neoplastic cellularity (mean 7.0). The percentage of cells in the phases S, G2 and M of the cell cycle by flow cytometry varied from 14 to 35 (mean 23%). Neither the cell ploidy nor the tumoral proliferative index were related with the survival of the patients (53 +/ 51 months). CONCLUSIONS: Lymphoma of the follicular mantle is a clinical pathological entity of low biologic aggressivity. Cell aneuploidy or ploidy and the tumoral proliferative index did not establish differences in prognosis or survival in this series. PMID- 8231355 TI - [Value of basal cortisol and ACTH in the immediate postoperative period of hypophyseal surgery in non-ACTH secretory adenomas]. AB - BACKGROUND: The aim of this study was to evaluate the usefulness of basal cortisol and ACTH in the immediate postoperative period of pituitary surgery as indicators of definitive adrenocorticotropin function. METHODS: Twenty-one patients with pituitary, non producers of ACTH, adenomas, three microadenomas and 18 macroadenomas treated by adenomectomy by a trans-sphenoidal route were respectively studied. The basal cortisol and ACTH were compared in the first week following surgery with the definitive results obtained after one month by dynamic tests (stimulation with ACTH or insulin hypoglycemia). RESULTS: The six patients with secondary adrenal failure (AF) in the definitive evaluation had lower basal cortisol in the immediate postoperative period than the patients with AF (135.3 +/- 225.3 nmol/l versus 473.6 +/- 147.2 nmol/l; p < 0.05). The values of ACTH were also lower (2.3 +/- 1.6 nmol/l versus 4.8 +/- 3.4; p < 0.05). In all the patients with definitive AF except one, the basal cortisol in the first week was lower than 130 nmol/l and in those who did not present AF it was greater than 220 nmol/l. CONCLUSIONS: In the immediate postoperative period after pituitary surgery cortisol is a good indicator of definitive adrenocorticotropin function. This parameter may identify the patients requiring posterior substitutive treatment. PMID- 8231356 TI - [Laparoscopic surgery. A revolution in general and digestive surgery]. PMID- 8231357 TI - [Malaria in Spain. Update]. PMID- 8231358 TI - [Liposome amphotericin in the treatment of deep mycoses in patients not severely immunosuppressed. An efficient alternative with low toxicity]. AB - Amphotericin is a powerful antifungal agent of high toxicity. Encapsulation in liposomes has led to new perspectives although clinical experience is still slight. Four patients, who were neither carriers of antibodies against the human immunodeficiency virus nor neutropenic, diagnosed of meningeal cryptococcosis, pleural aspergillosis, cerebral aspergillosis and ophthalmic candidiasis, respectively and treated with liposomal amphotericin are reported. The treatment was effective and well tolerated. Clinical improvement was observed in the patient with cerebral aspergillosis but magnetic resonance demonstrated persistence of the lesions. Only slight deterioration in renal function was observed in one case and in the other two renal failure improved upon substitution of conventional amphotericin by liposomal amphotericin. The slight systemic toxicity and the absence of local intolerance allowed the administration of high doses and shortening of the therapeutic schedule. PMID- 8231359 TI - [Lipoprotein(a): from hidden pre-beta lipoprotein to cardiovascular risk factor]. PMID- 8231360 TI - [Type I diabetes mellitus registries. A current necessity]. PMID- 8231361 TI - [Endogenous opioid peptides in septic shock]. PMID- 8231362 TI - [Tetany and foscarnet]. PMID- 8231363 TI - [Early detection of colorectal cancer in the workplace]. PMID- 8231364 TI - [Early detection of colorectal cancer in the workplace]. PMID- 8231365 TI - [Nosocomial sepsis caused by Xanthomonas maltophilia]. PMID- 8231366 TI - [Acquired aplastic anemia: a stereological analysis of bone marrow fatty tissue and its clinical correlations]. AB - BACKGROUND: To analyze the changes of the number and size of bone marrow adipocytes in aplastic anaemia. To discover possible clinical and haematological factors which may be associated with predominant expansion of the number or size of adipocytes. To analyze the prognostic value of the adipocyte number and size. METHODS: The adipose tissue of 25 plastic-embedded bone marrow specimens corresponding to patients with aplastic anaemia was studied stereologically. The correlation study between the results obtained and clinical and haematological data was carried out. RESULTS: The expansion of the adipose tissue is due to the predominant increase of the adipocyte number in some cases and of the adipocyte size in others. Among different clinical and haematological factors, only a lower age is significantly associated with a predominant increase of adipocyte number. In the prognostic stratification performed according to three different systems, neither the adipocyte number nor the size showed a statistically significant association. CONCLUSIONS: In the aplastic anaemia, the younger patients expand their bone marrow adipose tissue by increasing predominantly the adipocyte number, whereas in the older patients this occurs by predominantly increasing the adipocyte size (may be due to the decrease in proliferative potential). On the other hand, both the adipocyte number and size lack prognostic significance. PMID- 8231367 TI - [The evaluation of a noninvasive monitor to record ambulatory arterial pressure: the DIASYS 200 Novacor]. AB - BACKGROUND: The aim of this study was to validate the measurement precision of the portable automatic non invasive monitor of blood pressure-Novacor DIASYS 200. METHODS: A mercury sphygmomanometer was used as the reference measurer. To validate the DIASYS 87 persons (45 males, 42 females--age range 17-76 years) with a systolic blood pressure between 90-225 mmHg and diastolic blood pressure of between 64-149 mmHg were selected. RESULTS: The differences between the two apparatus were: systolic blood pressure 6.1 +/- 12.2 mmHg (r = 0.909); diastolic blood pressure -1.9 +/- 6.8 mmHg (r = 0.929) and cardiac frequency -2.0 +/- 6.3 ppm (r = 0.942). In healthy subjects of under 31 years of age (n = 20) the differences for systolic and diastolic blood pressure and cardiac frequency were 0.2 +/- 8.0 mmHg; -1.3 +/- 5.8 mmHg and -0.1 +/- 8.0 ppm, respectively. For subjects over the age of 59 years (n = 29) the measurements were 9.2 +/- 12.0 mmHg, 2.5 +/- 6.0 mmHg, -1.5 +/- 4.4 ppm, respectively. With the sample subdivided according to blood pressure the differences found in the normotensive subgroup (n = 23) were: -1.7 +/- 7.8 mmHg, -2. +/- 4.9 mmHg, and -2.4 +/- 7.9 ppm, respectively; in the subgroup with slightly high blood pressure (n = 40) 6.4 +/- 14.0 mmHg, -2.8 +/- 6.0 mmHg and -2.3 +/- 5.1 ppm, and in the subgroup with moderate-severe high blood pressure (n = 24) 10.4 +/- 10.9 mmHg, 0.2 +/- 9.1 mmHg and -0.9 +/- 6.5 ppm, respectively. CONCLUSIONS: The DIASYS fulfills the criteria of validation and precision for the measurement of diastolic blood pressure and cardiac frequency in all the cases. However criteria for systolic blood pressure are only fulfilled in normotensive and young subjects (less than 31 years of age). PMID- 8231368 TI - [The interdisciplinary model and multidisciplinary teams in geriatric care]. PMID- 8231369 TI - [The report to the court: the implications for physicians and patients. New medicolegal concepts of the misdemeanor and crime of injuries]. PMID- 8231370 TI - [Giant-cell myocarditis: a systemic disease? Apropos a case]. AB - Giant cell myocarditis is a rare disease of unknown etiology, which develops as a myocardial isolated affection or associated to different diseases. Its characteristics are such as necrosis, inflammation and giant cell presence in the myocardium. We present the case of a woman who suffered of giant cell myocarditis, thymoma, myasthenia gravis, chronic lymphocytic thyroiditis, giant cell myositis, granulomatous infiltration in the lymph nodes of the hilus of the lung and hypogammaglobulinemia; multiple association that we have not found in any published medical paper and that suggest the autoimmune origin of this illness. The cardiovascular symptoms and the associated diseases are revised, and we discussed the diagnostic and therapeutic topics, pointing out the necessity to take it into account for any patient with thymoma or myasthenia gravis developing to heart failure or arrhythmias. PMID- 8231371 TI - [Biomedical information systems or networks (I)]. PMID- 8231372 TI - [The optimization of the results by using thrombolytic therapy in acute myocardial infarct (and II)]. PMID- 8231373 TI - [Progressive neurological disorders in a 30-year-old male parenteral drug abuser]. PMID- 8231374 TI - [Herpes zoster during foscarnet treatment in an AIDS patient]. PMID- 8231375 TI - [The prevaccination determination of hepatitis B virus markers. Is their determination really necessary?]. PMID- 8231376 TI - [Reflections on the figure of the doctoral thesis advisor]. PMID- 8231377 TI - [2,3-dimercaptosuccinic acid: a new chelating agent, safe and efficacious, for lead poisoning]. PMID- 8231378 TI - [Nocturnal enuresis due to doxazosin]. PMID- 8231379 TI - [The use of antibiotics in surgical prophylaxis]. PMID- 8231380 TI - [Morbidity and mortality of osteoporotic proximal femoral fracture after one year follow-up]. AB - BACKGROUND: The aim of the present study was to follow for a year all the osteoporotic proximal femoral fractures that happened in the island of Gran Canaria during 1990 and to analyze: a) mortality; b) the degree of functional capacity; c) their destination after being attended from the fracture. METHODS: Personal interviews were performed in almost every patient (68%), either directly to them or the relatives who were at their care (23.5%), after excluding those patients who died during the acute phase (7.6%) a year after the fracture. Two patients were missed (0.9%). We had the collaboration of every public and private center in the island. 209 patients (154 women and 55 men) were interviewed. Age mean of the patients was 77.1 +/- 10.9 years. The degree of functional capacity was determined by Karnofsky scale. RESULTS: Mortality within the acute phase was 7.6%, rising to 20.8% 6 months later and to 30.7% after a year. There was an important disability in 30.7% of the remainder living patients. In the postsurgical period, 18% of the patients were transferred to a center for chronic care and 25% to a rehabilitation center. Although 48.9% of the patients are discharged to their homes from the hospital, surprisingly, when they die, only 15% do so in their homes, while 59% of the patients die in a center for chronic care. CONCLUSIONS: Osteoporotic proximal femoral fracture produces an important mortality within the first year after fracture, and also an important deterioration of functional capacity. Most of the patients are admitted later on and die in a center for chronic care. PMID- 8231381 TI - [Immunity coverage against measles, rubella and parotiditis viruses in a juvenile population in Leon, Spain]. AB - BACKGROUND: The immunity coverage, seroprevalence of IgG antibodies against infection by measles, rubella and parotiditis viruses in a juvenile population (50,398 children) was studied. METHODS: Systematic sampling was performed among children from 5-14 years of age who had undergone clinical analysis in the health care area of Leon. The hypothesis of sampling was the most unfavorable (p = q = 0.5) and the size of the sample of 600 children. Information was collected on vaccination state, previous history of disease and place of residence. The presence/absence of IgG antibodies was investigated by a commercial immunoenzymatic technique (EIA-Stat Measles, Mumps, Rubella, Whittaker Bioproducts, USA). RESULTS: Sixty percent of the children studied had IgG antibodies against the three virus, 27% against two and 9% against only one with absence of antibodies against the three virus in 3%. The seroprotection observed was significantly higher (p < 0.05) for measles (87% +/- 3%) that for the other two diseases (rubella 80% +/- 3% and parotiditis 77% +/- 3%). In the group of children with previous history of measles and parotiditis a higher percentage of antibodies (p < 0.01) was observed than in the group with previous history of vaccination (measles: 94% versus 84%, parotiditis: 90% versus 75%). No difference was observed in the case of rubella. CONCLUSIONS: Vaccination is still the principal conditioning factor of the state of immunity of the juvenile population in Leon in relation with the diseases studied (measles, rubella, parotiditis). The place of residence (rural or urban) did not condition different immunity coverage in the sample studied. The susceptibility of infection for some of these virus continues to be high: measles 13%, rubella 20% and parotiditis 23%). PMID- 8231382 TI - [Pulmonary infection in patients with lung cancer]. AB - BACKGROUND: To determine the frequency of pneumonia in patients with primary lung cancer requiring hospital admission, the etiologic diagnosis, probable production mechanisms, and the factors which may condition the evolution. METHODS: A prospective study was carried out in patients with primary lung cancer requiring hospital admission between 1986-1990. The diagnosis of pneumonia was performed by clinical and radiologic criteria and the etiology, in all cases, by hemocultures, pulmonary aspiration puncture, transtracheal aspiration and bronchoscopic techniques. RESULTS: The frequency of pneumonia was 12%. Etiologic diagnosis of the pulmonary infection was achieved in 16 patients and the most frequent etiologic agents were Gram negative (56%), anaerobic (31%) and Pneumocystis carinii (31%). Mortality was 16%. Community or nosocomial infection was not related with the evolution of the lung infection. Neither did diagnosis of the etiologic agent condition mortality attributable to the pneumonia but facilitated treatment. CONCLUSIONS: Pulmonary infection is a frequent and severe disease in patients with primary lung cancer. Bronchial obstruction may be the related pathogenic mechanism in the greatest number of cases with pneumonia. Due to the etiologic spectrum of these infections the practice of reliable diagnostic techniques is recommended to obtain etiologic diagnosis in addition to the administration of initial wide spectrum empiric antibiotherapy against the mentioned germs. PMID- 8231383 TI - [Compassionate therapy: an efficient and safe treatment?]. PMID- 8231384 TI - [New therapeutical uses of the interferons]. PMID- 8231385 TI - [Alpha interferon treatment of idiopathic myelofibrosis]. AB - Idiopathic myelofibrosis is a chronic myeloproliferative syndrome for which there is no effective treatment. The good results obtained with interferon in other chronic myeloproliferative syndromes have led their being tested in idiopathic myelofibrosis, but to date the experience is scarce. Four patients out of a total of 12 diagnosed with idiopathic myelofibrosis over a period of 3 years were selected for interferon treatment. Patients with low leukocyte or platelet counts or with contraindication for administration of the drug were excluded. Alpha-2b interferon was administered at an initial dose of 3 MU/day which was increased at 4-6 weeks to 5 MU/day in cases of insufficient response and if tolerance so permitted. In patients in whom favorable response was observed a maintenance schedule was initiated with low doses of interferon. Treatment was discontinued in two patients due to bad tolerance at 6 and 8 weeks of initiation of treatment with no response having been observed until that time. In the other patients favorable response was reported to interferon after 5 months of treatment with disappearance of the symptomatology, normalization of the hemo-peripheral values and a marked reduction of splenomegaly. This responses was accompanied by a decrease in bone marrow fibrosis in one case and total disappearance of the same in the other patient. Alpha-2b interferon constitutes an effective therapy for a selected number of patients with idiopathic myelofibrosis. Greater experience would allow the identification of the subgroup of patients who may benefit from this type of treatment. PMID- 8231386 TI - [The importance of the treatment of pain in thoracic trauma]. PMID- 8231387 TI - [Treatment and prophylaxis of the principal infections associated with the human immunodeficiency virus]. PMID- 8231388 TI - [Correction of resistance to erythropoietin after parathyroidectomy]. PMID- 8231389 TI - [Eosinophilic pleural effusion associated to bronchiolitis obliterans with organizing pneumonia]. PMID- 8231390 TI - [How much do you drink? sensitivity, specificity and predictive value]. PMID- 8231391 TI - [Frequent errors in acute carbon monoxide poisoning]. PMID- 8231392 TI - [Goodpasture's syndrome with antineutrophil cytoplasmic antibodies]. PMID- 8231393 TI - [Rheumatoid arthritis and hemiplegia: remission of the arthritis in the paretic limbs and appearance of a subcutaneous nodule in the non paretic elbow]. PMID- 8231394 TI - [The consumption of vitamins and sociocultural status]. PMID- 8231395 TI - [The usefulness of the bcr/abl rearrangement in the diagnosis and evolution of chronic myeloid leukemia]. AB - BACKGROUND: The rearrangement of the bcr/abl gene constitutes the molecular substrate of the Philadelphia chromosome (Ph'). The aim of this study was to analyze the usefulness of bcr/abl rearrangement in the diagnosis and evolution of chronic myeloid leukemia (CML). METHODS: The rearrangement of the bcr/abl gene was studied in 81 cases of which 34 corresponded to patients with CML (29 Ph' positive chromosome, 2 Ph' negative chromosome and 3 without karyotype), 2 patients with Ph' positive acute lymphoblastic leukemia, 15 patients with chronic myeloproliferative syndromes different from CML and 30 controls. Of the patients with CML, 6 were reevaluated when a blastic crisis was developed, 2 after receiving interferon treatment and 1 following allogeneic bone marrow transplantation. The technique used was that of Southern blotting using the restriction enzymes Bgl II and BamHI and the transprobe. RESULTS: Rearrangement of the bcr/abl gene was observed in all the patients with CML except in one with Ph' negative chromosome. In the remaining cases bcr/abl rearrangement was not observed. CONCLUSIONS: The Southern blotting technique for the study of the bcr/abl gene rearrangement is a sensitive and specific method in the diagnosis of chronic myeloid leukemia constituting a valid alternative to chromosomic study when this cannot be carried out or is not conclusive. It may also be used for the control of treatment in chronic myeloid leukemia. PMID- 8231396 TI - [Tuberculosis in a prison population: a study of 138 cases]. AB - BACKGROUND: Tuberculosis continues to be a disease of great importance in Spain with few data known on prison populations. In the present study an analysis of the incidence and characteristics of tuberculosis in the prison population people is done. METHODS: The General Penitentiary Hospital attends a prison population from the penitentiary centers of Madrid and its surrounding areas. From 1 March 1991-31 August 1992 a retrospective study of the patients with tuberculosis diagnosed by culture isolation of Mycobacterium tuberculosis was performed. Demographic, clinical, analytical, microbiologic analysis as well as data concerning to associated infections, antituberculous treatment and its possible toxicity as well as the evolution of the patients were collected. RESULTS: During the period studied 138 patients--120 corresponding to the area of Madrid and 18 to patients referred from other penitentiary areas--were diagnosed. The rate of incidence of tuberculosis in the penitentiary population in the area of Madrid was of 1170.5 cases per 100,000 prisoners per year (confidence interval [CI] 95%; 961-1380). The mean age was 30 years (CI 95%; 29-31) and 97% were males. Eighty four percent were positive for the human immunodeficiency virus (HIV) and 84% were intravenous drug addicts. The clinical form of presentation of the tuberculosis in the HIV positive patients was: pulmonary in 57% of the cases, disseminated in 29% and extrapulmonary in 14% and was pulmonary in all the cases of HIV negative patients. The mean CD4 lymphocytes count was 0.216 x 10(9)/l in HIV positive patients with pulmonary tuberculosis, and in disseminated tuberculosis was 0.062 x 10(9)/l (p < 0.001). The positivity of acid-alcohol resistant staining in the sputum samples and/or induced sputum was 67.4% in the patients with pulmonary tuberculosis and 77.4% in disseminated tuberculosis. Ten percent of the patients were found to abandon antituberculous treatment in the first trimester. CONCLUSIONS: The incidence of tuberculosis in the prison population in much higher than that for the general population in Spain, largely due to the high prevalence of HIV infection in this population. PMID- 8231397 TI - [Pregnancy and systemic lupus erythematosus]. AB - BACKGROUND: The interrelations between systemic lupus erythematosus (SLE) and pregnancy are reason for debate. Patients with SLE may present antiphospholipid antibodies (APL), anti-Ro (a-Ro) or anti-La (a-La) with their influence in pregnancy not being well defined. METHODS: The pregnancies in a series of 96 fertile age women with SLE, followed during the period of 1975-1991 with a mean follow up of 6.2 years were studied. The spontaneous or provoked abortions, prematurity and the number of flares of the disease during pregnancy and during the year after birth were analyzed. The number of flares was compared with that presented by a control group of women with SLE of similar clinical features followed over the same time period. RESULTS: Twenty-seven patients had 35 pregnancies with 23 full term pregnancies, 5 spontaneous abortions and 7 provoked abortions. Twenty-six percent of the deliveries were premature. One or two flares related with 43% of the pregnancies with viable fetus (11 flares in total) were registered. The number of flares in the control group was 6%. The difference was not statistically significant (p = 0.55) but the relative risk was of 1.6. No relation was found between the presence of APL, a-Ro and a-La and the course of the pregnancy. No case of lupus neonatorum was observed. CONCLUSIONS: Pregnancy is not advised in patients with systemic lupus erythematosus in remission. Patients should be more frequently controlled during pregnancy and post partum. The presence of antiphospholipid antibodies or anti-Ro does not modifies the prognosis. PMID- 8231398 TI - [Bacteremia due to Mycobacterium tuberculosis in patients coinfected with the human immunodeficiency virus]. AB - BACKGROUND: Since the appearance of AIDS, bacteremia by Mycobacterium tuberculosis is an event described with ever greater frequency. The isolation of M. tuberculosis in blood cultures in patients coinfected by the human immunodeficiency virus and M. tuberculosis may vary the initial clinical diagnosis and confirm the picture as disseminated tuberculosis. METHODS: The clinical records of patients admitted to the General Penitentiary Hospital of Madrid with significant bacteremia during the study period (March 1, 1991 September 1992) were retrospectively and prospectively reviewed. Special attention was given to the bacteremias by M. tuberculosis with the most significant clinical evolutive data being collected. The blood cultures were performed according to the routine technique using a non radiometric system with 10-day incubation and posterior processing of the sample for visualization and culture for mycobacteria. RESULTS: In an 18 moth period and out of a total of 284 positive blood cultures (154 significant isolations) 14 episodes of bacteremia by M. tuberculosis were studied in 12 prison patients coinfected by the human immunodeficiency virus with associated severe immunosuppression (mean of CD4 = 0.068 x 10(9)/l). All the patients were intravenous drug addicts. Likewise, 37 other significant bacteremias were reviewed. The positivity of the blood culture as the sole sample or associated to respiratory isolations, changed the initial diagnosis and classified the picture as disseminated tuberculosis on 6 occasions (43%). All the patients presented pictures of disseminated tuberculosis although only four had been previously diagnosed of AIDS. CONCLUSIONS: Bacteremia by M. tuberculosis is frequent in patients infected by the human immunodeficiency virus. M. tuberculosis (27.4%) was the most frequently isolated microorganism in blood cultures in this study. The processing of blood cultures for mycobacteria may be useful in patients positive for the human immunodeficiency virus with tuberculosis in order to confirm the diagnosis of disseminated tuberculosis. The isolation of M. tuberculosis in the blood may vary the initial diagnosis and could modify the prevalence of AIDS cases in Spain. PMID- 8231399 TI - [Molecular biology studies in chronic myeloid leukemia]. PMID- 8231400 TI - [Hypertension and stress]. PMID- 8231401 TI - [New prospects in the treatment and prevention of nosocomial pneumonias]. PMID- 8231402 TI - [Ductal carcinoma in situ of the breast]. PMID- 8231403 TI - [Compression neuropathy of the sciatic nerve during anticoagulant treatment]. PMID- 8231404 TI - [Poisoning by delayed-release verapamil in renal insufficiency patients]. PMID- 8231405 TI - [Jaundice and thrombolytic agents in patients with acute myocardial infarct]. PMID- 8231406 TI - [Bronchiolitis obliterans with organized pneumonia secondary to gold salts]. PMID- 8231407 TI - [Abscess of the right psoas coexistent with septic oligoarthritis]. PMID- 8231408 TI - The history of tuberculosis as a global epidemic. AB - TB should be thought of as a slowly progressing worldwide epidemic. Initially it was a disease of lower mammals, and the etiologic agent probably preceded the development of man on earth. It became an uncommon endemic disease in man about the time man began to settle in villages and develop agriculture. Crowding in European cities, and later the industrial revolution in Europe, provided the necessary environmental conditions for the endemic disease to become epidemic. For the next 400 years, the disease was spread by European empire-building and colonization. It came late to sub-Saharan Africa and to the Pacific Islands, and still later to the highlands of New Guinea. The epidemic gradually wanes within a large population group as resistant individuals survive and reproduce. This natural resistance is reflected in the ability of the macrophage to control intracellular growth of the organism. The resistant host shows a chronic infection primarily affecting the lungs, whereas the highly susceptible host shows a rapidly fatal illness with generalized spread of disease to many organs. Survivors of the initial infection then show another type of resistance to reinfection that is based on sensitized T cells. When this system is only partially successful, the host becomes infectious and capable of spreading the infection widely. The study of the epidemiology of TB and the evaluation of various public health measures to prevent or contain the disease requires that the investigator have an understanding of the nature and duration of the TB epidemic in the particular population under study. This factor is a much greater determinant of the course of an epidemic than any public health measure that man can institute, just as the currents in a river can have a more powerful effect on the course of a canoe than the most vigorous paddler. PMID- 8231409 TI - Immunology of tuberculosis. AB - TB is a chronic, necrotizing infection caused by M. tuberculosis. The clinical manifestations of disease are the result of a balance between the host response and bacterial virulence. Cellular immunity is responsible for effective control of infection, but cytokines released during the process of cellular immunity may also cause harm to the host. Humoral immunity plays little part in protection against TB. Individuals with defective cellular immunity are much more susceptible to disease from M. tuberculosis and are more likely to have a disseminated form of TB. PMID- 8231410 TI - Pharmacology of the antimycobacterial drugs. AB - The management of MDR-TB requires that the clinician become familiar with the "second-line" antimycobacterial agents. These drugs are generally less potent and frequently more toxic than isoniazid and rifampin. Because they are less active, innovative dosing schedules may allow us to take advantage of the few strengths that they possess. This approach will require further research into the dose response relationships for each agent. Based on our current knowledge of these drugs, practical guidelines for their use have been described. These guidelines include the gradual escalation of the oral doses of PAS, cycloserine, and ethionamide over several days, and the intravenous administration of streptomycin and capreomycin. Both ciprofloxacin and ofloxacin may be used for the treatment of MDR-TB, but data from clinical trials are currently lacking. Finally, because patients with AIDS appear to develop antimycobacterial drug malabsorption over the course of their HIV infection, therapeutic drug monitoring can be used to verify drug absorption in the individual patient. This approach may improve therapy for that patient and prevent the selection of additional drug resistance. PMID- 8231411 TI - Preventive therapy for tuberculosis. AB - TB is the most prevalent infection in the world. The emergence of the HIV epidemic has dramatically increased the risk of developing clinical TB in infected persons, substantially increasing TB rates globally. Preventive therapy with INH for 6 to 12 months substantially reduces the risk of developing clinical TB in persons with tuberculous infection. Yet, preventive therapy is not fully used because of lack of knowledge about TB among health care professionals, patient nonadherence with preventive therapy regimens, and drug toxicity. Persons at high risk for developing TB should be screened for infection with a tuberculin skin test and given preventive therapy when appropriate. Promising new regimens of shorter duration and new drug delivery systems are currently being evaluated. Preventive therapy must be widely adopted if the goals of TB control and eventual elimination are to be achieved. PMID- 8231412 TI - The treatment of tuberculosis. AB - Short-course chemotherapy has made the treatment of TB easier and better than ever, but it works only when patients take the drugs regularly. Compliance is a must for therapy to be successful. Physicians treating patients with tuberculosis should be acutely aware of noncompliance, and every effort to ensure adequate treatment must be put forth. Directly supervised therapy is an excellent option when enough resources are available. Intermittent regimens markedly reduce the manpower required for observed therapy. New agents are being tested for in vitro activity against M. tuberculosis, and clinical studies of those found to be potentially effective are needed to formulate new regimens against the ever increasing threat of multidrug-resistant TB. PMID- 8231413 TI - Compliance with tuberculosis therapy. AB - The treatment experience in TB control can be a very rewarding time for both the patient and the provider. If the patient is made to feel that the provider cares for him as a person, a meaningful relationship can develop and can have a positive influence on the patient's behavior in taking medications and perhaps even in the direction of his life. Noncompliance with medications unfortunately has been a "given" in the TB control program. The successful program must be patient centered and include the following components: (1) fair and equal treatment of all patients, (2) patient involvement in the plan of care, (3) the most current patient treatment and services, including the availability of DOT for patients with TB infection and disease, and (4) rewards for patients and staff for positive behavior in the completion of treatment. Given current levels of staffing and other resources, some noncompliance will continue even in the best TB programs. Caring and knowledgeable staff members who support the patient centered approach can make TB treatment a positive experience for both the patient and themselves. PMID- 8231414 TI - The role of the public health department in tuberculosis. AB - The role of the public health department in TB is a critical component of the overall TB control effort. This article illustrates both the traditional public health methods of surveillance, containment and prevention, and some of the newer strategies being employed to address TB control in today's multifaceted environment. It shows that controlling TB will require an intensification of collaborative efforts between public, private and community providers. In particular, the role of public health and health care workers in institutional settings is emphasized as it relates to shared community efforts. In light of the recent outbreaks of drug-resistant disease and the associated dramatic increasing TB morbidity and mortality, the need for these partnerships is urgent. Given the legal mandate for TB control, health departments will continue to play a major role in the elimination of this disease. The deterioration of these public health services, however, will require immediate attention lest the very foundation of TB control be allowed to crumble. PMID- 8231415 TI - Environmental control of tuberculosis. AB - The premise of this article is that it is possible to reduce but not eliminate the risk of TB infection in many institutional settings through environmental interventions--as supplements to conventional public health TB control efforts. Of the environmental means available to reduce the concentration of infectious droplet nuclei, ventilation, isolation strategies, and personal respirators have received the most attention, but each has inherent limitations. Ventilation and other air-moving strategies (i.e., fan-filter and fan-UV room units) are limited by the large volume of air that must be moved to dilute and remove already dilute droplet nuclei. Isolation assumes that potential transmitters are suspected, whereas negative pressure in isolation rooms is difficult to achieve and maintain in many hospitals. The use of well-designed small enclosures for sputum induction and other high-risk procedures, however, should provide highly effective source control. Personal respirators have a limited protective role because they cannot be worn by all workers at all times, and cannot reasonably be issued to other patients and visitors. Germicidal UV irradiation of upper room air is widely misunderstood, but offers practical air disinfection that can be safely and efficiently deployed in a variety of high-risk environments. Although there are theoretical and experimental bases for these recommendations, there are no clinical field trials preventing TB using any of the available environmental interventions, primarily because of the highly variable nature of TB transmission. PMID- 8231416 TI - Tuberculosis in children. AB - The dramatic resurgence and increase in the total number of cases of tuberculous infection and disease in children is alarming in the United States. With poverty, poor access to health care, overcrowding (predominantly in inner-city areas), and an increase in immigration from areas with high endemic rates of TB, the problem in children will continue to increase. If the impact of coinfection with HIV and M. tuberculosis becomes significant in children, as it has in adults in the United States, the increase in the total number of cases of tuberculous disease in children could be staggering. The impact of multidrug-resistant strains of M. tuberculosis and the current crises in availability of effective anti-TB drugs will need a similar resurgence. PMID- 8231417 TI - Tuberculosis in the elderly. AB - Elderly persons of today are survivors of a time when most adults were infected with tuberculosis. Although many people died at that time, others survived the initial infection but continued to harbor viable bacilli in dormant caseous and calcified lesions. This article discusses the tuberculin test in the elderly, transmission and risk of infection in nursing homes, clinical manifestations, diagnosis, management considerations, preventive therapy, and surveillance of tuberculosis in nursing homes. PMID- 8231418 TI - Tuberculosis in patients with HIV infection. AB - Tuberculosis is the most common opportunistic infection in patients with HIV infection worldwide and is the only one that is transmissible to others by the respiratory route. Tuberculosis is curable and preventable. Early detection of tuberculosis disease and infection in individuals with or at risk for HIV infection is paramount. This approach can minimize the devastating interaction between these two diseases. PMID- 8231419 TI - The epidemiology of multidrug-resistant tuberculosis in the United States. AB - There has been a significant increase in the number of cases of MDR-TB in the United States. Although cases of MDR-TB have been reported from many areas of the country, the majority of the cases are concentrated in large urban areas. MDR-TB is difficult and expensive to treat. CDC has developed a National Action Plan to Combat Multidrug-Resistant Tuberculosis. The main elements of this plan include (1) greater surveillance and epidemiologic studies of drug-resistant TB; (2) initiatives to make the laboratory diagnosis of MDR-TB more rapid, sensitive, and reliable; (3) education of health care professionals about MDR-TB, its prevention, control, and treatment; and (4) measures to facilitate the development of new antituberculous drugs. CDC has published guidelines for the prevention of nosocomial spread of MDR-TB. to prevent the development and spread of MDR-TB, medical practitioners must suspect TB and make the diagnosis as rapidly as possible. Once a patient is diagnosed with TB, the most important step to prevent the development of drug-resistant disease is to ensure that patients take all of their medication. Directly observed therapy is the best way of ensuring this. In addition, more specific interventions, such as the use of incentives to improve compliance in certain situations, may need to be applied to groups in which high rates of drug resistance have been found, such as HIV positive persons, IDUs, homeless persons, and persons who have been exposed to persons with MDR-TB. Quick and effective public health interventions targeted at these defined groups should help to control the spread of both drug-susceptible and drug-resistant TB. PMID- 8231420 TI - PDRD--a computer-based primary care decision support system. AB - This article identifies the current stage of development and assessment of computer-assisted decision support systems in the domain of general practice. Physician's Desk Reference and Database (PDRD), an electronic medical database, is presented, and a plan for assessment of PDRD is briefly discussed. PMID- 8231421 TI - A statistically rule-based decision support system for the management of patients with suspected liver disease. AB - The paper describes how a decision support system in liver diseases, mostly oriented to prediction of the necessity for liver biopsy, has been developed. The system designed is a hybrid one and consists of two parts: logical and statistical. The logical part contains rules, formulated on the basis of current medical knowledge, which enables recognition of clear cases; diseased or non diseased. The unclear cases are classified on the basis of rules statistically extracted from databases. These rules have been reached after a comprehensive exploratory analysis of the sample of 165 patients with slightly to moderately raised levels of routine liver tests but without signs or symptoms of liver diseases. The extracted decision diagrams which simulate traditional medical diagnosis conduct have been found to be superior to discriminant analysis and probabilistic inductive learning. They use only a limited number of laboratory tests to detect the necessity for biopsy. PMID- 8231422 TI - Decision support for drug prescription integrated with computer-based patient records in primary care. AB - A conceptual model of an information system that integrates a controlled vocabulary, a patient database, and a knowledge base is described. Methods, design and components for the implementation of the system are discussed. It is argued that the key issue for the successful introduction of computer-based decision support in primary care today is integration with a computer-based patient record. Also important is that the knowledge acquisition process is based on the general practitioner's real needs. This has been achieved by, first, providing general practitioners with real patient data from a series of retrospective database studies; and second, letting a panel of general practitioners select, discuss and decide which computer reminders to implement. A hybrid representation scheme was chosen for the knowledge base. The combination of a standard procedural representation (the so-called Arden syntax) for the reminder knowledge with a semantic net representation for the medical factual knowledge facilitates knowledge sharing with other systems and knowledge reuse within the system. PMID- 8231423 TI - Unimage, a new RIS for the DIOGENE 2 environment at Geneva Cantonal University Hospital. AB - The effective management of a radiology department depends on adequate computing tools for imaging specialists--radiologists and technicians--and administrators. The rapid evolution of imaging techniques in radiology requires flexible systems for describing the daily operation of a service and accessing data generated by every possible source. In order to compensate for the insufficiencies of our current radiology information system (RIS), and within the framework of the migration towards the DIOGENE 2 hospital information system, a new RIS, named Unimage, has been implemented at the Cantonal University Hospital of Geneva. Unimage has been designed not only to fit the current and future needs of the radiology department, but also to operate in different services, i.e. diagnostic radiology, nuclear medicine and therapy radiology. Among its functionalities, Unimage offers a simple two-way data communication with the PACS world. The technical choices we have made in the design of the new RIS include a distributed environment, as well as a graphical (X/Motif) user interface. The main functionalities and the technical implementation of Unimage are described in this paper. PMID- 8231424 TI - Case-based reasoning algorithms applied in a medical acquisition tool. AB - The knowledge of clinical medicine can, efficiently and effectively, be elicited and encoded automatically as cases. The developed knowledge base often contains some redundancy. The cost can be tolerated as it affects neither the performance of the system nor the integrity of the knowledge. The advantages are simplicity in development and reliability in performance. PMID- 8231425 TI - Sleep Expert--an intelligent medical decision support system for sleep disorders. AB - A new type of associative knowledge-based decision support system (Sleep Expert) for the diagnosis and classification of sleep disorders is described. Sleep Expert is based on the International Classification of Sleep Disorders (1990). The programming system used was KnowledgePro (Windows), a high-level language that integrates object-oriented programming, hypertext and expert system technologies. Sleep Expert is an interactive program composed of 288 separate integrated submodules and 264 text files. The program includes eight reasoning questions about symptoms setting the limits for the diagnosis subset. The user obtains a list of possible diagnoses on the screen where he/she can examine their criteria. The program has been written in such a form that the user can freely associate and can move forwards and backwards. Detailed information is included in hypertext. PMID- 8231426 TI - Validation of a metabolic prototype to assist in the treatment of insulin dependent diabetes mellitus. AB - This paper describes the principles and prototyping of a computer system to assist in the treatment of patients with insulin-dependent (type 1) diabetes mellitus. The system adopts a mixed approach involving rule-based qualitative algebra and a dynamic mathematical model to define the relationships between insulin dosage, diet and glycaemic response. The rule-based system (KBS), implemented in PROLOG, can be used to generate qualitative therapeutic advice. These suggestions are quantified and rank-ordered by the use of a mathematical model of glucose-insulin interaction in type 1 diabetes mellitus, with parameters adjusted for individual patients. In this paper an overview of the integrated prototype, linking the KBS and model, is provided and a case study used to demonstrate the principles of the system in operation. The results of verification and validation work performed on the KBS are described. PMID- 8231427 TI - [Chemiluminescence of granulocytes in response to Pseudomonas strains isolated from various materials]. AB - The study was aimed at comparison of chemiluminescence of granulocytes during their incubation with suspensions of Pseudomonas strains isolated from various sources. The study was performed on 136 strains isolated from clinical material. The results were correlated with resistance of these bacteria to bactericidal activity of serum and their ability to produce proteolytic enzymes. It was found that strains isolated from blood and pus weakly activate granulocytes in contrary to bacteria isolated from urine and feces. Ability to pronounced activation of granulocytes was without relation to susceptibility of a given strain to action of serum. But it correlated negatively with ability to production of proteolytic enzymes. These observations indicate existence of relation between characteristics of strain and type of clinical material from which it was isolated. PMID- 8231428 TI - [Evaluation of drug sensitivity and biochemical properties of coagulase-negative S. aureus strains isolated from clinical specimens]. AB - 20-25% of strains isolated in our hospital in 1991 from clinical specimens and identified as S. aureus were coagulase-negative. These strains were characterized in respect of biochemical properties and resistance to antibacterial drugs. It was shown that the investigated group of strains displayed high drug resistance and particularly high percent of strains were resistant to methicillin (60%). 100% strains were resistant to penicillin and tetracyclines and most of them were resistant to aminocyclitol antibiotics. Coagulase-negative strains, in comparison with coagulase-positive, less frequently produced hemolysins and more frequently staphylokinase. PMID- 8231429 TI - [Susceptibility in vitro to certain quinolines of gram-negative bacteria and gram positive cocci causing urinary tract infections]. AB - Susceptibility to norfloxacin, ofloxacin, pipemidic acid and nalidixic acid of 848 bacterial strains isolated from urine of patients treated in 1989-1992 in Wroclaw hospitals was investigated. The study, performed by the disc diffusion methods, concerned 568 Enterobacteriaceae strains, 147 Gram-negative non fermenting bacteria and 133 strains of staphylococci. Highest percentage (90 100%) of susceptibility to all used antimicrobial agents was detected among Escherichia, Proteus, Morganella and Citrobacter. Less frequent susceptibility (30-70%) was observed among Klebsiella, Enterobacter and Serratia. Among strains of P. aeruginosa susceptible to norfloxacin and ofloxacin were, respectively, 61.4 and 22.2% isolates. Over 95% of strains of P. aeruginosa were resistant to nalidixic acid. Among other non-fermenting Gram-negative bacteria, over 50% were resistant to norfloxacin and ofloxacin. Both S. aureus and S. epidermidis were susceptible to ofloxacin and norfloxacin in 81-93% of tested strains. They were 2 3 times less frequently susceptible to pipemidic and nalidixic acid. PMID- 8231430 TI - [Susceptibility of microorganisms to ofloxacin]. AB - The authors have compared susceptibility of bacteria isolated from various clinical material in 1989-1991 to ofloxacin. Staphylococcus aureus, Escherichia coli, Proteus, Klebsiella, and Enterobacter were frequently susceptible to the antibiotic, the percentage of resistant strains being less than 10-20%. A higher percentage of resistance was noted for Pseudomonas aeruginosa--approx. 40%. A gradual increase in resistance to ofloxacin among Streptococcus faecalis, Serratia and Acinetobacter strains was observed. PMID- 8231431 TI - [Bacterial flora in laryngeal carcinoma and in postoperative scars of patients treated prophylactically by ciprofloxacin]. AB - Microbiological investigations regarded samples from diseased tissue in patients with larynx carcinoma and in 53 patients. Samples were also taken two weeks after surgery from postoperative wounds. The patients received after surgery ciprofloxacin and metronidazole for five days. Cultures were incubated in aerobic and anaerobic conditions. Isolated microorganisms were tested by application of the API system susceptibility to ciprofloxacin, netilmicin and gentamycin were determined. Samples from cancerous tissue and postoperative wounds revealed most frequently presence of streptococci, staphylococci, Bacteroides and intestinal bacteria. It was found, however, that in investigated patients different bacterial flora was present when compared with previously tested cancerous tissue. Hemolytic streptococci and Staphylococcus aureus, and less frequently enterococci, were found much more frequently, Isolated strains were susceptible to ciprofloxacin (93%), netilmicin (76%) and gentamycin (39%). PMID- 8231432 TI - [Investigations concerning the influence of cefuroxime and amikacin on dynamics of selected parameters of nonspecific cellular immunity in experimental animals]. AB - Rabbits divided into two groups were injected intramuscularly antibiotics amikacin or cefuroxime in doses corresponding to mean therapeutic doses for seven days. Blood samples for determination of values of phagocytosis reaction (percentage of phagocytizing neutrophils, phagocyte index) and NBT test were taken before application of antibiotics and on 7, 14, 28 and 42 day of study. After application of cefuroxime significant decrease of NBT value was observed at the 14th day, whereas the percentage of phagocytizing neutrophils was changing at 7 and 14 day. Phagocytic index did not change. After application of amikacin, percentage of phagocytizing granulocytes was significantly decreased (7, 14 and 28 day). Phagocytic index did not change. NBT test value was lowered at 14 day, but it increased at the 42 day. PMID- 8231433 TI - [Papilloma virus infections in women with cervical pathology]. AB - The presence of Papilloma capsid antigen in patients hospitalized in the Institute of Gynecology and Obstetrics was studied, using immunoperoxidase technique. Of 73 patients, 44 had cervical intraepithelial neoplasia (CIN), in the following grades - CIN 1 (n = 21), CIN 2 (n = 5), CIN 3 (n = 18). Two cases of squamous cell carcinoma were confirmed. In 9 other patients cervical infections were found. Eighteen women with normal PAP smears were treated as the control group. For Human Papilloma Virus studies, smears were taken from 73 patients, and cervical from 8 women only. Of the 44 patients with CIN, in 7 cases (15.2%) capsid antigen was found. The frequency of Papilloma virus antigen detection was higher in the biopsy material than in the cervical smears. Chlamydia trachomatis infections and the presence of Mycoplasma and Ureaplasma in the genital tract were also studied. Chlamydia trachomatis was found in 14.3% of the CIN patients, and Ureaplasma urealyticum dominated both in women with CIN (40.5%) and in women with genital inflammatory diseases (55.6%). PMID- 8231434 TI - [Cytomegalovirus infections in infants examined in the department of virology in 1988-1992]. AB - Active cytomegalovirus (CMV) infections were found in 91 infants (11%) examined in the Department of Virology from 1988 to 1992. Serological methods (ELISA, CFT) and virus isolation from urine were used to diagnose CMV infection. Congenital or perinatal infections were recognized in 20 infants, while 10 infants were infected by blood transfusion. In most cases the route of CMV transmission was not recognized. The occurrence of clinical symptoms in CMV-infected children was also studied. In 80% of the infants the disease was characterized by hepatosplenomegaly or liver dysfunction. CMV pneumonia was found in 41% of the cases, and gastroenterocolitis in 36% of the subjects. PMID- 8231435 TI - [Bacteriophages of serologic group A converting synthesis of staphylokinase and beta toxin in S. aureus]. AB - The properties of the eight S. aureus bacteriophages of the serogroup A converting staphylokinase production were investigated. Three of them were able to a double conversion: production of the staphylokinase and inhibition of beta toxin synthesis. All of the investigated bacteriophages were classified as the I morphological group of the Styloviridae on the basis of the electron microscope analysis. The size of capsids of the examined bacteriophages was 77 +/- 2.8 nm x 43.1 +/- 1.9 nm and the tail length was 272.7 +/- 12.7 nm. Most of them (6 bacteriophages) had the tail terminated in the basal plate. The lytic properties of the investigated bacteriophages were not identical. Seven of them showed features of the III and one of the V (miscellaneous) lytic group. PMID- 8231436 TI - [Increase of pathogenicity of Staphylococcus aureus strains caused by lysogenic conversion by phages of serologic group F]. AB - The substantial influence of presence of the double converting prophages of the serological group F on the decrease of intensity of human leukocytes stimulation with lysogenic cells of S. aureus was determined. The end values of chemiluminescence in the six experimental systems containing lysogenic bacteria were only from 15.4 to 37.2% of the value in the control sample. The end values of the intercellular killing of the two lysogenic variants were twofold lower than in controls. Third of the investigated variants was completely resistant to intracellular killing in the applied system, and revealed 27% of growth stimulation. PMID- 8231437 TI - [Influence of antagonistic flora on quantity of staphylococci in the oral cavity]. AB - The study was aimed at establishment, whether in oral cavity co-inhabit bacteria demonstrating antagonistic properties against staphylococci and whether this phenomenon may influence the size of staphylococcal population in this environment. Two locations of bacteria in oral cavity were investigated--saliva and tooth plaque. It was found that total quantity of staphylococci (both coagulase-positive and/or coagulase-negative) in saliva decreases with the age of investigated persons. Mean values amount respectively for children to 3.5 +/- 1.0 log CFU/ml and for adults to 2.9 +/- 0.7 log CFU/ml. This is statistically different (p = 0.0003). Amount of staphylococci in tooth plaques remains generally at the same level and amount to 1.4 +/- 0.6 log CFU/ml. In each tested sample of saliva and tooth plaque, presence of bacteria antagonistic to isolated from a given sample staphylococci, was detected. Among 2-4 year children and in adults, percentages of antagonistic bacteria against coagulase-positive staphylococci were higher than against coagulase-negative strains. Among children aged 5-6 and 11-13 years opposite proportions were found. Statistic analysis confirmed existence of negative correlations, both linear and ranking, between quantity of antagonistic bacteria and amounts of coagulase-positive and coagulase negative staphylococci in the same tested samples of saliva and tooth plaque. These results suggest that antagonism between bacterial flora of oral cavity and staphylococci, is presenting one of factors limiting the quantity of staphylococci in this environment. PMID- 8231438 TI - [Production of glycocalyx by coagulase negative staphylococci isolated from the oral cavity]. AB - The study was aimed at determination whether and how frequently coagulase negative staphylococci persisting in human oral cavity possess the same feature. The material consisted of 255 strains of coagulase-negative staphylococci isolated from oral cavity of persons of both sexes and in differing age. Glycocalyx production by these strains was tested quantitatively by a colorimetric method. Glycocalyx was produced by 222 out of 225 tested strains (87.1%). For individual genera this frequency was following: epidermidis 164/186, hominis 14/14, warneri 12/13, saprophyticus 7/8, auricularis 4/5, haemolyticus 1/4, simulans 4/4, caseolyticus 2/2, xylosus 0/1 and for unidentified 14/18. Majority of tested strains produced only slight amounts of the glycocalyx. Only 46 out of the -20.7% of active strains (18.0% of all tested strains) produced large amount of glycocalyx. Among these strains, 37 belonged to the genus epidermidis, 6 to hominis and one each to warneri, simulans and auricularis. There was no correlation between amount of glycocalyx produced by individual strains and their number in the oral cavity. These results indicate that production of glycocalyx is a common feature of coagulase-negative staphylococci belonging to different genera and may appear with different intensity. Oral cavity may be a source of strains producing large amounts of glycocalyx and thus potentially pathogenic. PMID- 8231439 TI - [Antagonism between oral cavity streptococci and staphylococci]. AB - The subject of this study was investigation of quantitative relations between streptococci, lactic acid bacilli and actinomycetes and staphylococci present in the oral cavity. It was found that in each tested sample of saliva streptococci are present which inhibit growth of an indicator strain Staphylococcus aureus 209P. Percentage of such streptococci varies from 11.1% to 100% (mean value 52.9 +/- 19.7%). There is a negative correlation between number of streptococci and staphylococci in samples of saliva delivered from the same oral cavity. Pearson's linear correlation coefficient was r = -0.7962 (p < 0.0001), and Spearman's correlation rank was rs = -0.7667 (p < 0.0001). Lactic acid bacilli were inhibiting growth of staphylococci only in conditions in which there were not neutralized by organic acids produced by these microorganisms. Actinomycetes had no influence at all on growth of these microorganisms. These results suggest that streptococci are responsible for antagonistic properties of oral cavity bacterial flora in relation to staphylococci. Numbers of streptococci conditions size of staphylococcal population in this environment. Staphylococci practically exert no antagonistic activity against cohabiting with them streptococci. PMID- 8231440 TI - [Modulation of lymphocytes and immunoglobulins by serine proteinase of Staphylococcus aureus]. AB - Serine proteinase of S. aureus binds with surface of peripheral blood lymphocytes and by proteolysis of polypeptides on the membrane and transduction of signals, changes in polyclonal activation of T and B lymphocytes occur. Cleavage of immunoglobulins tested in three classes and evaluated by electrophoresis in polyacrylamide gel (SDS-Page) of Ig fragments, was differing. IgG and IgG fragments were degraded in higher degree than IgM, H and L chains or i Fc and i Fab fragments. Influence of proteolytic modulation of IgG particles on fixation antigenic fractions homogenized by ultrasound cells of S. aureus was evaluated in ELISA test. Basing on detailed results postulated mechanism of modification effect of immunoglobulins and lymphocyte reaction under influence of serine proteinase of S. aureus in microenvironment of after-inflammatory reaction in staphylococcal infection, has been accepted. PMID- 8231441 TI - [Regulatory influence of Staphylococcal serine protease on selected functions of lung macrophages in vitro. Introduction of cellular carbonic paramagnetic probe method for evaluation of cellular phagocytic activity]. AB - The study was aimed at evaluation of properties of serine protease produced by Staphylococcus aureus V8. Influence of enzyme on chemotactic and phagocytic activity of rat lung macrophages was investigated. Chemotactic activity of rat lung macrophages was studied by application of modified Boyden chambers. Slight induction of chemotaxis of cells preincubated with enzyme was found. Phagocytic activity of rat lung macrophages was investigated by application of Varian E3 spectrometer. EPR signals emitted by paramagnetic graphite phagocytized by macrophages were measured. Increased phagocytic activity of macrophages subjected to preincubation with enzyme in concentration of 10 micrograms/ml was observed. It seems that staphylococcal proteinase is engaged in interaction of staphylococci with immunological system cells. PMID- 8231442 TI - [Occurrence of antituberculous antibodies in patients with lung tuberculosis and other inflammatory states of the respiratory system]. AB - In this study an indirect immunofluorescence method was applied. Determinations regarded the level of anti-H37Rv antibodies in patients with bacteriologically confirmed lung tuberculosis and in a control group consisting of healthy volunteers. Antibody levels were significantly higher in sick patients. We conclude that this method may be applied as a supplementary diagnostic test in doubtful cases and that it can prove useful as screening test for investigation of groups of population especially at risk from tuberculosis. Applied method of indirect immunofluorescence is easy to perform, quick (time of the test takes about 3 hours), available in Poland, and cheaper than ELISA. PMID- 8231443 TI - [Weakness of cellular response to Listeria antigens in pregnant mice]. AB - It was found that pregnant mice exhibit increased susceptibility to lethal action of Listeria monocytogenes, although there was no correlation between increase of susceptibility of animals to lethal effect of bacteria and weakening of elimination of microorganisms from the spleen. Increased susceptibility of animals to listeriosis was accompanied by weakening of some parameters of cellular immunity. In pregnant mice of strains C57Bl/6, naturally resistant to infections, inhibition of II-1 by macrophages was found which at the same time exhibited depression of chemotaxis. Weakening of these macrophage functions did not influence the development of delayed hypersensitivity to Listeria antigen (LA) in C57Bl/6 mice--the reaction which was suppressed in pregnant mice of A/J strain which is characterized by natural susceptibility to listeriosis. Inhibition of development of delayed hypersensitivity to LA in A/J pregnant mice was caused by defective function of dendritic cells and T lymphocytes. Dendritic cells of A/J pregnant mice exhibited limited ability of presentation of LA to immune T lymphocytes isolated from animals infected with L. monocytogenes or L. innocula. On the other hand, immune T cells of A/J mice cultured with LA exhibited inhibition of proliferation and production of MIF. PMID- 8231444 TI - [Protective action of anti-listeria IgG]. AB - It was found that single injection into mice of 70 micrograms IgG against Listeria innocua protected them against lethal action of virulent cell of L. monocytogenes, if antibodies were injected 1-3 weeks before infection. Immune IgG did not exert the protective effect when injected one day before infection. Mice T lymphocytes receiving immune IgG three weeks before infection, exhibited increased ability to direct interactions (cluster formation) with dendritic cells presenting Listeria antigen (LA). At the same time, sera obtained from these animals were blocking the reaction of indirect haemagglutination occurring between antibodies against L. innocula and LA-covered sheep erythrocytes. These results suggest that anti-listerial IgG are inducing in mice production of anti idiotypic antibodies. Most probably, some of these immunoglobulins imitate determinants of Listeria antigens, copying their three-dimensional structure. They could activate T cells specific against bacterial antigens (1st signal of activation). Introduction of virulent L. monocytogenes cells into mice possessing T cells activated by anti-idiotypic antibodies, could create a second antigenic signal, resulting in intensive elimination of bacteria by mechanisms of specific cellular immunity. PMID- 8231445 TI - [Interaction of cefotaxime and normal human serum in the bacteriocidal process against strains of Escherichia coli producing surface K1 antigen]. AB - A comparison was made regarding susceptibility of strains of E. coli to bactericidal activity of normal human serum. It was found that when among strains with K1 antigen over 40% were resistant to normal human serum, strains without this antigen were resistant in only 15%. Sub-doses of cefotaxime (1/2 and 1/3 MIC) together with normal human serum strong bactericidal action against strains of K1-possessing E. coli resistant to serum. Sub-doses of cefotaxime cause occurrence of elongated form of bacteria, starting one hour after addition of the antibiotic. Elongated forms of bacteria are not loosing antigen K1 and this suggest that it probably does not exert a role in protection of bacteria against synergistic action of both studied factors. PMID- 8231446 TI - [Further types of core lipopolysaccharides in Proteus]. AB - Comparative analysis of the chemical composition of 11 core oligosaccharides isolated from lipopolysaccharides of the wild (S) and phenotypically rough (R) strains Proteus mirabilis (nine) and Proteus vulgaris (two) allowed to recognize three new types Proteus core, classified as IV, V, VI. All of them contained D galactose and D-galactosamine in addition to common core constituents: D-glucose, D-galacturonic acid, L-glycero-D-manno-heptose, KDO, EtN described for Proteus core types I, II, III (6, 7, 8). D-glucosamine was characteristic for Proteus core type VI whereas D-glycero-D-manno-heptose for types V and VI. PMID- 8231447 TI - [Selected properties of strains of a new species of Proteus penneri from the second American collection]. AB - The second collection of the novel species Proteus penneri consists of 25 strains from which only two have shown rough from properties in the tests differentiating S and R variants of bacteria. The migration pattern of their lipopolysaccharides in gel electrophoresis was leader-like, typical for smooth organisms. 13 out of 25 lipopolysaccharide preparations showed strong-reactivity with anti-0 sera in semi-quantitative precipitation test. Serological similarity between the strains within species Proteus penneri is discussed. PMID- 8231448 TI - [Production of staphylokinase and hemolysin by coagulase-negative staphylococcus]. AB - The ability to staphylokinase production by the representative of the six of fifteen investigated species of staphylococci was detected: S. epidermidis, S. lentus, S. sciuri, S. lugdunensis, S. xylosus and S. hominis. The frequency of occurrence of this feature was different among the investigated species. Relatively, the least frequently this feature was observed with S. epidermis (2.3%) and S. xylosus (8.3%) strains. This property most frequently occurred among S. lentus--all the examined strains of this species produced staphylokinase. The hemolysins synthesis was shown among 13 of 15 investigated coagulase-negative species of staphylococci. The only two species of the examined representatives did not produce any hemolysin: S. hyicus and S. schleiferi. Complete correlation between staphylokinase synthesis and the absence of beta hemolysin production was observed among strains: S. lugdunensis and S. lentus as with the lysogenic S. aureus strains with the double converting phages. PMID- 8231449 TI - [Isolation and certain biological and immunologic properties of Proteus penneri strains from the European collection]. AB - The American Collection of 45 Proteus penneri strains was supplemented by 22 strains isolated in Poland and Germany. All strains exhibited typical smooth forms of bacteria. Susceptibility to antibiotics was determined and compared with the other P. penneri strains. 0-antigenic relatedness and serological similarity between strains in the whole Proteus penneri collection were discussed. PMID- 8231450 TI - [Characterization of hemolytic activity of Proteus penneri]. AB - Bacteria belonging to the genus Proteus synthesise two kinds of hemolysins HpmA and HlyA which represent "RTX proteins". In previous papers we described the production of an extracellular HlyA hemolysin by some P. penneri strains. Now we are reporting on the synthesis by P. penneri, typical for P. mirabilis HpmA hemolysin. There were identified two P. penneri strains 5 and 37 in which both hpmA and hlyA regions are present. In two other strains P. penneri 13 and 44 only hlyA region was found, whereas in strain P. penneri 42 operon hpmA was identified. The production of HpmA hemolysin was revealed in the cases of P. penneri 5, 42 and P. mirabilis 03 and 1959. The dynamics of HlyA hemolysin synthesis by P. penneri 44 was also investigated and its highest activity was observed during logarithmic phase of growth of bacterial culture. HlyA hemolysin was isolated from culture filtrate by precipitation with polyethylene glycol 4000. The invasiveness of HpmA+ and/or HlyA+ P. penneri strains was also checked by use of mouse L929 fibroblasts. Both kinds of strains were able to penetrate tested cells. The invasion of L929 fibroblasts by strains producing HlyA hemolysin is accompanied by cytotoxic effect. PMID- 8231451 TI - [Immunochemical studies of O-specific polysaccharide from Proteus penneri 14 lipopolysaccharide]. AB - O-specific polysaccharide was obtained on mild acid degradation of Proteus penneri strain 14 lipopolysaccharide and found to contain equimolar of D galactose, D-ribose, 2-acetamido-2-deoxy-D-glucose, N-(D-galacturonoyl)-L-alanine and 3-(Nacety-L-alanyl)-amido-3,6-dideoxy-D-glucose. On the basis of non destructive NMR analysis it was concluded that repeat unit of the 0-specific polysaccharide of P. penneri 14 has the following structure: -2-beta-D- Quip3NAlaAc-(1-->4)-alfa-D-GalpAAla-(1-->2)-beta-D- Ribf-(1-->4)-beta-D-Galp-(1- >3)-beta-D--GlcpNAc-(1--> This structure was confirmed by structural elucidation of trisaccharide and disaccharide fragments prepared on mild acid hydrolysis of the polysaccharide. Immunodominant role of the partial structures of the pentasaccharide repeating unit in manifesting serological specificity of P. penneri 14 was discussed. Very weak cross-reactions of P. penneri anti-serum were observed with E. coli 0114 and Shigella boydii 08 LPS's, which showed some structural similarities. No cross-reaction with P. mirabilis 027 LPs was detected. PMID- 8231452 TI - [Immunochemical studies of O-specific polysaccharide of Proteus penneri 42 lipopolysaccharide]. AB - O-specific polysaccharide was obtained an mild acid degradation of Proteus penneri strain 42 lipopolysaccharide and found to contain D-glucose, D galacturonic acid and 2-acetamido-2-deoxy-D-glucose in molar ratio 2:1:1. Methylation analysis showed that the polysaccharide is linear, one of the glucose residue is substituted at position 2, the second one and the residue of galacturonic acid at position 4, and the 2-acetamido-2-deoxy-D-glucose residue at position 3. On the basis of non-destructive NMR analysis the following structure of repeat unit of the O-specific polysaccharide was established and confirmed independently by methylation analysis: -2)-beta-D-Glc-(1-->4)-beta-D-Glc-(1-->3) beta-D-GlcNAc-(1-->4)-al fa-D-GalA-(1--> The serological investigation with application of P. penneri strain 42 anti O-serum has shown the activity of homologous preparations of LPS and PS, as well as cross-reactions with heterologous lipopolysaccharides from other Proteus strains. PMID- 8231453 TI - [Structural and immunologic studies of Proteus mirabilis 033 O-specific polysaccharide]. AB - O-specific polysaccharide was obtained by mild acid degradation of P. mirabilis 033 lipopolysaccharide (LPS). It was found to contain N-acetyl-D-glucosamine, D glucuronic acid and N-acetyl-L-fucosamine in a ratio 1:1:1. On the basis of the data obtained from 13C-NMR and methylation analysis, the following structure of repeating unit was established: [formula: see text] Selective removal of the D GlcA significantly decreased reactivity of 033 O-specific polysaccharide with homologous antiserum. This component was plays an immunodominant role. Cross reactivity between anti-033 serum and disaccharide alfa-L-FucNAc-beta-D-GlcNAc containing P. vulgaris 023 and S. arizonae 059 O-specific polysaccharides was also observed. PMID- 8231454 TI - An in vitro method using vascular smooth muscle cells to study the effect of compounds on cell proliferation and intracellular lipid accumulation. AB - Primary cultures of vascular smooth muscle cells (SMC) were incubated with hyperlipidemic rabbit serum (HLRS) to produce intracellular lipid accumulation. Cultures were exposed to different concentrations of the test compounds and their effects upon cell proliferation and intracellular lipid accumulation were measured. The in vitro results obtained with different Ca2+ antagonists, a HMG CoA reductase inhibitor and other hypolipidemic substances showed a strong correlation with the in vivo activities of these compounds, suggesting that cultured SMC may be a useful initial in vitro model to detect new chemical entities active upon atherosclerosis related parameters. PMID- 8231455 TI - ICI 206,970: a novel eukalemic diuretic with calcium channel blocking activity. AB - ICI 206,970, an aminomethylphenol pyrazine derivative, produced diuretic and saluretic effects, but caused only minimal alterations in kaliuresis in dogs and rats after oral and parenteral administration. ICI 206,970, unlike HCTZ, increased diuretic activity in a more clearly defined dose-related manner, and did not reach the plateau level even up to a dose of 100 mg/kg p.o. The diuretic and natriuretic response is believed to be a combination of a direct effect on tubule function and changes in GFR, particularly at higher doses. Based upon studies in in vitro amphibian models for mimicking mammalian nephron, ICI 206,970 appeared to possess a furosemide-like activity in toad cornea and amiloride-like activity in toad bladder. It is concluded that ICI 206,970 is a potent eukalemic diuretic with the potential of multiple sites of renal action. PMID- 8231456 TI - Studies on the effects of endothelin-1 (ET-1) and endothelin-3 (ET-3) in brain hypoxia and on the participation of brain prostanoids in their actions. AB - The effects of endothelin-1 (ET-1) and endothelin-3 (ET-3) in brain hypoxia have been studied in mice using the following experimental models: hypobaric hypoxia induced by low atmospheric pressure, histotoxic hypoxia induced by 12.5 mg/kg KCN i.p., and complete ischemia induced by decapitation. ET-1 and ET-3 were injected intracerebroventricularly (i.c.v.) 15 min before the tests. Forebrain tissue concentrations of 6-keto-PGF1 alpha and thromboxane B2 (TxB2) were measured 15 min following i.c.v. administration of ET-1 (5 pmol/mouse) and ET-3 (10 pmol/mouse). ET-1 (1-5 pmol/mouse) and ET-3 (5-25 pmol/mouse) showed a dose dependent increase in the survival/gasping time in all models of hypoxia. The effect reached its maximum between 15 and 30 min after ET administration and lasted for about 120 min. ET-1 and ET-3 did not significantly change the brain levels of 6-keto-PGF1 alpha and TxB2. The protective effect of ET-1 and ET-3 was unexpected, because endothelins (ETs) are the most potent vasoconstrictors known, and in doses close to those used in this study they cause vasoconstriction and decrease in cerebral blood flow. The protection was not likely to be due either to stimulation of the endogenous release of prostacyclin (PGI2) or to a decrease in the deleterious prostanoid thromboxane A2 (TxA2). Additional experiments are necessary to explain the cerebroprotective effects of ET-1 and ET-3. PMID- 8231457 TI - Comparison of a conscious versus anesthetized canine model in response to milrinone, hydralazine and nitroprusside. AB - The purpose of this study was to demonstrate, based on previous and new data, that the differences between a conscious and anesthetized canine model in the cardiovascular responses to cardiovasoactive agents were beyond their difference in the sensitivity of the compensatory mechanisms. In both conscious and anesthetized canine models, mean arterial pressure (MAP) was decreased by hydralazine (at 1-3 mg/kg and 0.3-3 mg/kg; by -26.5 +/- 4.5 and -18.8 +/- 11.7% [max. changes expressed as mean +/- SEM], respectively). MAP was also decreased by nitroprusside (both at 0.01-0.1 mg/kg, by 54.6 +/- 2.8 and -60.5 +/- 3.0%, respectively) in the conscious and anesthetized models. However, the differential MAP responses to hydralazine and nitroprusside between the two models are inconsistent with a difference in the sensitivity of the two models. Hydralazine at 1 mg/kg decreased MAP greater in the conscious than anesthetized model, whereas nitroprusside decreased MAP similarly in the two models. In conclusion, not all differential responses to hydralazine and nitroprusside between conscious and anesthetized canine models can be explained by a difference in the sensitivity of their compensatory mechanisms. PMID- 8231458 TI - DNA purification by polycarbonate filters. AB - Organic solvent free procedures are described for the purification of mammalian DNA from rat liver, kidney, spleen, lung and brain. The basis of the purification procedures are the use of the detergent sodium dodecyl sulfate (SDS) and inert polycarbonate filters with 2 mcm pores which retain long macromolecules such as DNA while allowing other cellular constituents to pass through the pores. Neither proteinase K nor use of peristaltic pumps were found to be absolutely required for DNA purification, although it is desirable to use peristaltic pumps to control solution flow rates. Of the 12 different procedures (1.2-4.2 h required) evaluated, purified mammalian DNA contained from 0 detectable to 1.5% of the original protein remaining as a contaminant. With the 4 recommended procedures, the average of 8 experimental values of DNA to protein ratios was 1.81. Depending on the diameter of the individual filter, about 12-90 mcg of rat liver DNA can be purified from each separate tissue or cell sample. PMID- 8231459 TI - The alpha study: multiple regression of the inositol 1,4,5-triphosphate signal transduction mechanism in burn trauma. AB - The novelty of applying 3-dimensional graphic capabilities, involving area and vector changes was used to understand variations in inositol derivatives due to the systemic effects of large body surface area (BSA) burns. This report is an attempt to broaden current perspectives on how such changes in inositol forms impact on the disposition of IP3 within skeletal muscle cells. Because it is the first of its type to evaluate systemic effects in this way, it is called the alpha study. Consideration of multiple factors (viz., 5 orders of magnitude) involved in burn trauma in this manner provides new insight into the pharmacologic changes which utilize the IP3 signal transducing system that underlie burn trauma. Such insight would prove beneficial in improving the quality of rehabilitative burn care with respect to skeletal muscle physiology. PMID- 8231460 TI - Transdermal patch delivery of acetylcholinesterase inhibitors. AB - Transdermal delivery of cholinesterase inhibitors (ChEI) for treatment of dementia would have advantages associated with continuous dosing and enhanced compliance, but feasibility depends on achieving desired levels of central nervous system enzyme inhibition. We developed a patch technique for assessing delivery of ChEI in rats and examined two organophosphate compounds, metrifonate and DDVP, and a carbamate, heptylphysostigmine, for production of peripheral and central nervous system ChE inhibition at target levels. With DDVP, a log dose/percent brain AChE inhibition was obtained over a range of 10-65% inhibition within a 10-fold concentration of inhibitor in the patch. Brain cholinesterase was inhibited up to seven days after a 24-h patch application. Long-term inhibition was greater than that attained after intramuscular injection, but without the rapid initial inhibition peak seen with the latter route. In contrast to DDVP, sustained high levels of brain enzyme inhibition could not be produced by transdermal delivery of metrifonate or heptylphysostigmine. Apparently DDVP has features, i.e., liquid state in pure form and high inhibitor potency, which make it particularly suitable for patch administration. PMID- 8231461 TI - Measurements of serum ACE activity in vitro after administration of enalapril in man cannot reflect inhibition of the enzyme in vivo. AB - We previously demonstrated the dependency of serum angiotensin converting enzyme (ACE) inhibition by the inhibitor enalaprilat in vitro on the concentration of substrate and enzyme present in the assay, according to a competitive tight binding mechanism. In the present study, the relevance of these findings for ex vivo measurements after administration of enalapril has been confirmed in serum samples of four patients which were assayed twice using different substrate concentrations in vitro. The measured extent of ACE inhibition in the samples was markedly different depending on the substrate concentration added in vitro (in relation to its Km value: S/Km, respectively), suggesting lower inhibition over time when higher S/Km was used. When the dilution factor and the added S/Km were taken into account the measured values could successfully be predicted from the respective serum enalaprilat concentrations by means of the concentration-effect model previously evaluated for the in vitro relationship (Emax tight model). Considering conditions which probably better reflect the situation in vivo (no dilution, physiological substrate concentrations far below Km) we simulated a time course of in vivo serum ACE activity in these patients which suggests almost complete inhibition of serum ACE over time in contrast to the in vitro measurement. Thus, we conclude that the usual ex vivo measurements of ACE activity lead to an underestimation of the extent of inhibition because of sample dilution and high exogenous substrate in vitro, and therefore must fail to reflect enzyme inhibition in vivo. PMID- 8231462 TI - [An American sociologist about Swedish health care: why this sudden confidence in the market?]. PMID- 8231463 TI - [Laboratory medicine and hospital laboratories. Effective activities when market is functioning]. PMID- 8231464 TI - [Don't the principle of selection and drop-out rate play any role?]. PMID- 8231465 TI - [Economic values are not the only controls]. PMID- 8231467 TI - [Structural formulas are necessary in Fass]. PMID- 8231466 TI - [Overtime work is not free]. PMID- 8231468 TI - [Shortage of pediatric radiologists]. PMID- 8231469 TI - [Vitamin A in retinitis pigmentosa. An effective treatment, for the first time]. PMID- 8231470 TI - [Vitamin A and retinoids. From natural products to potent drugs]. PMID- 8231471 TI - [Physician/patient consultation in migraine treatment]. PMID- 8231472 TI - [Treatment of acute migraine with sumatriptan. Clinical experiences with advantages and disadvantages]. PMID- 8231473 TI - [After stroke. Psychological and social changes for the patient and family]. PMID- 8231474 TI - [Removal of allergens helps children with asthma]. PMID- 8231475 TI - [Mammography screening. A pilot study of the psychological effects]. PMID- 8231476 TI - [Clinton's health care reform: private health insurance but public cost control]. PMID- 8231477 TI - [The EEC creates a European Medical Product Agency]. PMID- 8231478 TI - ["P-marked" against one's own will]. PMID- 8231479 TI - [Breast reconstruction--but not for all cases]. PMID- 8231480 TI - [Clinical social medicine more and more important]. PMID- 8231481 TI - [Is the physician in charge of epidemics control forgotten?]. PMID- 8231482 TI - ["Giraffe language" for understanding of feelings/needs]. PMID- 8231483 TI - [Geriatric care is improving]. PMID- 8231484 TI - [Simplified handling of aminoglycosides. Daily dosage established on basis of serum creatinine levels]. PMID- 8231485 TI - [Laparoscopic surgery. Knowledge of pneumoperitoneum and anesthesia can prevent perioperative complications]. PMID- 8231486 TI - [Malignant mesothelioma. New diagnostic and therapeutic methods make the disease curable]. PMID- 8231487 TI - [Subarachnoid hemorrhage. The most serious disease of the stroke group]. PMID- 8231488 TI - [High frequency of suicide among younger physicians. Unsatisfactory working situations should be dealt with]. PMID- 8231489 TI - [Mentally ill persons are a problem in home care services. Better personnel training is needed]. PMID- 8231490 TI - [Recruitment of physicians for the UN battalion in Bosnia is difficult]. PMID- 8231492 TI - [24-hour-scheduling--is it so bad?]. PMID- 8231491 TI - [Are the drug committees sold?]. PMID- 8231493 TI - [The sting method in the treatment of ureteral reflux]. PMID- 8231494 TI - [Why this anxiety about euthanasia?]. PMID- 8231495 TI - [Standard of living and health among the elderly. Not only the social class is important]. PMID- 8231496 TI - [A talk with the national epidemiologist Margareta Bottiger. Prevention against infections through 5 decades. Interview by Bo Lennholm]. PMID- 8231497 TI - [Drug abuse among immigrants. A problem with different aspects]. PMID- 8231498 TI - [Class and morbidity among the oldest old. A study shows a general connection]. PMID- 8231499 TI - [Surveys of standard of living among the oldest old. Important to choose the right test and the right interviewer]. PMID- 8231500 TI - [Aurora. Help to women who are afraid of labor]. PMID- 8231501 TI - [New psychiatric law in Russia. In certain aspects more far-reaching than the Swedish one]. PMID- 8231502 TI - [Early diagnosis is important in ovarian cancer]. PMID- 8231503 TI - [Children are losers in the African AIDS epidemic. Protective network of the extended family is diminishing]. PMID- 8231504 TI - [Antidepressive agents with only few serious adverse effects]. PMID- 8231505 TI - [Profit interest will be the ideal of market-oriented health services. Monopoly can function in a democratic way]. PMID- 8231506 TI - [False care about the rights of HIV- infected patients]. PMID- 8231508 TI - [District general practitioners are suppressed in the discussion of the family practice reform]. PMID- 8231507 TI - [Unemployment and the 3rd way]. PMID- 8231509 TI - [We want to take care of a healthy relative]. PMID- 8231510 TI - [Role of the magnetic camera in injuries of the cervical spine. Should be available day and night in the future]. PMID- 8231511 TI - [Undergraduate medical education. Are more of the humanities and critical thinking needed?]. PMID- 8231512 TI - [Diabetes mellitus type 1 with onset between ages 15-34 years. Report of current status from the study of diabetes incidence in Sweden]. PMID- 8231513 TI - [Bird fancier's alveolitis. An underdiagnosed disease?]. PMID- 8231514 TI - [Medical students about current undergraduate medical education: no stimulation of critical thinking]. PMID- 8231515 TI - [The patient in ambulatory surgery. Inadequate information and insufficient analgesia are the most frequent complaints]. PMID- 8231516 TI - [A case report. Torsades de pointes caused by overdose of thioridazine]. PMID- 8231517 TI - [Demands--control--support. A new model for research of occupational environment]. PMID- 8231518 TI - [The criminally insane in prisons. Radical measures are needed]. PMID- 8231519 TI - [Facts about concealed trauma care!]. PMID- 8231520 TI - [Ambulances are part of the health services]. PMID- 8231521 TI - [Should we accept defective copies?]. PMID- 8231522 TI - [Do patients under oxygen therapy have to be prisoners of their own communities?]. PMID- 8231523 TI - [Documented effect of the morning-after pill. New elements in contraception counseling]. PMID- 8231524 TI - [Not only the orienteers. A study of mortality from myocarditis among the younger Swedish population]. AB - All cases of death in the 11-45-year-old age group, occurring during the period 1981-1986, where myocarditis was given as the underlying or contributory cause, were analysed. Estimated per 100,000 person years, mortality was 0.5 for men and 0.2 for women. There was a tendency toward geographic clustering and, independently, a clustering in time. Some variation according to socio-economic class was also present. We conclude that the current focus of Swedish research on myocarditis among orienteers is too narrow, as more than 90 percent of cases occur outside this group. PMID- 8231525 TI - [Chlorhexidine prophylaxis at labor. Prevention of sudden infant death?]. AB - In a study of nearly 5,000 births it was reported that decontamination of the birth canal at labour with chlorhexidine prevents excess neonatal morbidity associated with group B streptococci. Toxigenic E coli or chlamydia, most likely transmitted from vagina to infant at labour, has been uncovered in victims of the sudden infant death syndrome (SIDS). Assuming an etiological connection, vaginal decontamination at labour might also reduce mortality in SIDS. In a study of co twins of SIDS victims, 13/23 survivors were reported to have long-term respiratory disorders. Recent reports link chlamydia with asthma and other respiratory dysfunction. This suggests an etiological connection between chlamydia, SIDS and respiratory dysfunction including asthma. It would be interesting if these hypothesized connections could be tested in an extension of the chlorhexidine study beyond infancy. PMID- 8231526 TI - [The woman in the scientific study--does she exist?]. PMID- 8231527 TI - [Little known radiation accidents in Russia. Radiation-induced leukemia and cancer]. PMID- 8231528 TI - [Epidemiology and tropical medicine. Stimulating education in London]. PMID- 8231529 TI - [Physicians' occupational conditions. High workload. Impossible to organize own work]. PMID- 8231530 TI - [The peer review system inhibits creative research. Let the scientists take care of the "risk capital"]. PMID- 8231531 TI - [Inadequate financing system for doctoral candidates. Routine research is supported--but not creativeness]. PMID- 8231532 TI - [Rectal cancer surgery, where and by whom?]. PMID- 8231533 TI - [Should a bulging Douglas' abscess not be surgically treated any longer?]. PMID- 8231534 TI - [Misleading information about vaccination against influenza]. PMID- 8231535 TI - [Can general practitioner take over the check-up done by the physician employed in occupational health services?]. PMID- 8231536 TI - [Osteoporosis can be iatrogenic--spare the ovary]. PMID- 8231537 TI - [Contact with reality is questioned]. PMID- 8231538 TI - [Incorrect about fat in meat]. PMID- 8231539 TI - [Resuscitation at any price?]. PMID- 8231540 TI - [National cooperation in mass vaccination of infants is important]. PMID- 8231541 TI - [What ambulance-based service is best for the patient?]. PMID- 8231542 TI - [Streptococcal myositis. A life-threatening soft tissue infection requiring early surgical intervention]. PMID- 8231543 TI - [New and old methods for the determination of renal function. More effective treatment requires better examinations]. PMID- 8231544 TI - [Prior to surgery of children with congenital heart defects. Examination without heart catheterization]. PMID- 8231545 TI - [Streptococcal myositis. The patient survived thanks to massive therapeutic measures]. PMID- 8231546 TI - ["Bodylink"--a new aid in diabetes education]. PMID- 8231547 TI - [Influenza A Beijing is approaching. 2 new strains in this season's vaccine]. PMID- 8231548 TI - ["Medecins sans frontieres", physicians without frontiers, now also in Sweden]. PMID- 8231550 TI - [What happens with us, future general practitioners?]. PMID- 8231549 TI - [Don't extend sale of the non-prescription drugs. There is a risk of diminished respect, increased number of poisoning cases]. PMID- 8231551 TI - [Continuing cooperation with psychiatric care in Latvia]. PMID- 8231552 TI - [No to unnecessary treatment of ureteral reflux]. PMID- 8231553 TI - [To be understood--not only refugees have the right]. PMID- 8231554 TI - [Why not more than one general practitioner?]. PMID- 8231555 TI - [Dogmatics and scolding]. PMID- 8231556 TI - [Appeal! Reports of adverse reactions of food are wanted!]. PMID- 8231557 TI - [Physicians must take over health care]. PMID- 8231558 TI - [Coloscopy is the most reliable method in suspected colonic disease]. PMID- 8231559 TI - [The general practice reform is a large-city reform]. PMID- 8231560 TI - [Answers to 10 questions. Passive smoking and coronary disease]. PMID- 8231561 TI - [Neonatal respiratory insufficiency. Milestones in therapeutic development]. PMID- 8231562 TI - [Surfactant therapy of premature infants. Complications and mortality are reduced]. PMID- 8231563 TI - [More effective treatment with the new CPAP technique]. PMID- 8231564 TI - [Respirator therapy of newborn infants]. PMID- 8231565 TI - [Severe epilepsy. Interdisciplinary cooperation improves diagnosis and treatment]. PMID- 8231566 TI - [Malignant lymphoma. Experiences with autologous bone marrow transplantation in 50 patients]. PMID- 8231567 TI - [Benzodiazepine findings in autopsy material. A study shows interacting factors in fatal cases]. AB - The paper consists in a report of a retrospective study (of data from 1987) on the prevalence of benzodiazepines in blood at the time of death. Of 2,007 autopsies, forensic chemical analyses were performed in 1,587 cases, in 159 of which benzodiazepines were found. Of these 159 deaths, 22 were considered to be due to natural causes, and in another 22 cases the cause of death was still unclear after examination; the remaining 115 deaths were due to accidents (N = 16), suicide (N = 60), drug addiction (N = 29) or alcoholism (N = 10). Multiple benzodiazepine intake was found in 37 cases, a subgroup including all 29 cases of death due to drug addiction. In a comparison of suicides and natural deaths, the concentrations both of flunitrazepam and nitrazepam were significantly higher among the suicides (P < 0.001 and P < 0.05, respectively). In four cases, the sole cause of death was benzodiazepine intake. It is concluded that some benzodiazepines, particularly flunitrazepam, may be more toxic than formerly supposed. PMID- 8231568 TI - [A case report. ACE blockade is hazardous in acute sodium loss]. PMID- 8231569 TI - [Ethical debate on preventive medicine]. PMID- 8231570 TI - [Preventive measures don't have high priority]. PMID- 8231571 TI - [Rehabilitation of vision disorders in Brazil based on the Swedish model]. PMID- 8231572 TI - Rebound caloric nystagmus. AB - Rebound caloric nystagmus (RCN), a distinctly different phenomenon from secondary phase nystagmus (SPN), was serendipitously discovered during the course of routine neurotological evaluations. RCN occurred when the horizontal semicircular canal was brought into a truly horizontal plane after the primary phase nystagmus (PPN) had ended. The maximum slow-phase velocities (SPVMax) of both PPN and RCN were almost identical. The pathological lesion was an Arnold-Chiari malformation (ACM). Two other patients with posterior fossa lesions also showed RCN. To determine if RCN is an unrecognized normal phenomenon, 11 healthy subjects were tested. The average SPV of RCN in normals was 0.7 degrees per second. Such a small value of RCN is probably of no significance. It therefore appears that RCN is a sign of posterior fossa neuroaxial lesions. PMID- 8231573 TI - Differences in dendritic cells in congenital and acquired cholesteatomas. AB - Cholesteatomas are histologically benign, though biologically invasive lesions that arise from the migration of squamous epithelium of the ear. Acquired cholesteatomas usually arise in an antigenically active environment, i.e., a chronically and/or recurrently inflamed middle ear. In contrast, congenital cholesteatomas occur in an uninflamed environment. The potential role of dendritic cells (DCs) in the evolution of this lesion has not been thoroughly studied. By staining for S-100 protein, the authors evaluated the presence and distribution of DCs in cholesteatomas. Sixteen cases of cholesteatomas diagnosed from 1987 to 1989 were selected for this study. The formalin-fixed, paraffin embedded sections were processed by a standard avidin-biotin peroxidase antiperoxidase method for S-100 protein and for leukocyte common antigen (LCA). The presence and distribution of S-100 protein-positive DCs was evaluated and compared to canal wall skin. DCs were present in all cases. Nine acquired cholesteatomas had 5 to 16 epithelial DCs per high-power field (HPF). Seven congenital cholesteatomas were examined. Four with isolated congenital pearl-like cholesteatomas had 1 to 3/HPF epithelial DCs. In contrast, the three inflamed congenital cholesteatomas had 6 to 12/HPF DCs. The control uninflamed canal wall skin had only 1 to 3/HPF DCs. All DCs were LCA negative, as expected. PMID- 8231574 TI - Motor innervation of the eustachian tube muscles in the guinea pig. AB - The brainstem location of motoneurons innervating eustachian tube-associated muscles in the adult guinea pig was determined using intramuscular injections of the neural tracer horseradish peroxidase (HRP). Following HRP injections into the tensor veli palatini and the eustachian tube belly of the medial pterygoid muscle, an ipsilateral column of HRP-labeled motoneurons was present medial to the dorsolateral division of the trigeminal motor nucleus. Following HRP injection into the levator veli palatini, labeled motoneurons were present in the ipsilateral dorsal division of nucleus ambiguus. The locations of the tensor veli palatini and levator veli palatini motoneurons are similar to those found in studies of other animals. A distinct eustachian tube belly of the medial pterygoid muscle was also identified. This sub-belly had a motoneuron pool distinct from the main medial pterygoid muscle group. The authors have provided the gross anatomical and neuroanatomical substrates upon which future studies of eustachian tube function in the guinea pig may be based. PMID- 8231575 TI - Configuration of the glottis in laryngeal paralysis. I: Clinical study. AB - It is widely accepted in the American literature that the cricothyroid muscle is responsible for the paramedian position of the vocal fold in recurrent laryngeal nerve paralysis. However, support in the literature for this theory is not conclusive, and the cadaveric vocal fold position expected after lesions of the vagus nerve has also been reported in patients with an intact superior laryngeal nerve. This study compares the configuration of the glottis in patients with unilateral paralysis due to known lesions of either the recurrent laryngeal or vagus nerve. Normal subjects were studied as controls. Results indicate that the alteration of glottic configuration in laryngeal paralysis cannot be adequately characterized by standard terms of vocal fold position. The paralyzed vocal fold is shortened, with anterior rotation of the arytenoid. Patients with vagus nerve lesions had a statistically insignificant tendency for a more lateral vocal fold position, but a discrete difference in position between the two groups was not identified. This study also confirms prior observations that paralyzed vocal folds are frequently not denervated. These findings have significant implications for management of laryngeal paralysis. PMID- 8231576 TI - Configuration of the glottis in laryngeal paralysis. II: Animal experiments. AB - It is widely believed that in isolated recurrent laryngeal nerve paralysis, the paralyzed vocal fold assumes a median or paramedian position, due to the action of the cricothyroid muscle. A review of the literature reveals that support for this theory is not conclusive and, in particular, experiments indicate that the cricothyroid muscle does not appreciably affect vocal fold position in acute paralysis. The research in this study compares the configuration of the glottis in chronic unilateral recurrent laryngeal nerve paralysis in cats, with and without concomitant denervation of the cricothyroid muscle. Results indicate that vocal fold position is not determined by the cricothyroid muscle. Incomplete denervation of intrinsic laryngeal muscles as well as synkinetic reinnervation appear to be significant factors in determining vocal fold position in chronic laryngeal paralysis. PMID- 8231577 TI - Effects of acid on the larynx of the maturing rabbit and their possible significance to the sudden infant death syndrome. AB - Sudden infant death syndrome (SIDS) has been shown to result from a variety of causes. One group of neonates at high risk for SIDS includes those who develop apnea secondary to gastroesophageal reflux (GER). Reflux has been shown to produce apnea in infants, and aggressive treatment results in significant improvement in symptoms. Because it is a site of resistance in the airway, the larynx plays an important role in the development of apnea. Through its sensory innervation, the larynx also serves as the afferent limb for reflexes that regulate respiration. In order to investigate the relationship between obstructive apnea and central apnea induced by the instillation of acid on the larynx, simulating GER, a rabbit model was developed. Maturing rabbits at 15-day intervals up to 60 days of age were studied using saline and acid solutions. Acid solutions produced obstructive apnea in all age groups. With acid solutions, central apnea occurred in all age groups but had a peak incidence at 45 days. Gasping respirations were seen in all groups but were most common at 30 days of age. Although obstructive and central apnea occurred together as mixed apnea, both types of apnea were seen independently of each other. Acid instilled on the larynx of maturing rabbits resulted in significant obstructive, central, and mixed apnea. Gasping respirations and frequent swallowing were frequent associated symptoms. Acid-induced obstructive apnea in rabbits mirrors symptoms seen in human infants with GER. Central apnea in infants with GER is seen less commonly; however, central apnea as the result of laryngeal stimulation has been demonstrated repeatedly in several animal models. Central apnea, culminating in fatal asphyxia, has been described in several animal models. The larynx appears to play a pivotal role in the development of apnea in susceptible infants with GER. PMID- 8231578 TI - The role of fenestrated vessels for the secretory process in the nasal mucosa: a histological and transmission electron microscopic study in the rabbit. AB - Both nasal glands and nasal vessels are supposed to be responsible for the humidification of inhaled air. After previous studies on the vasculature of the nasal septal mucosa in the rabbit, we examined vessels of the anterior concha in order to find a morphological equivalent to explain the role of endonasal vasculature in the secretory process of this tissue. The main findings of our study include the detection of fenestrated endothelial walls of both capillaries and venous sinuses located underneath and directly adjacent to the basal cell layers of the epithelium. The same phenomenon-fenestrated vessels underneath the epithelium with the fenestrated parts facing the epithelial cells-has been described in other parts of the body and has been called "polar differentiation." In accordance with the interpretations of other authors, we conclude that these features are responsible for the regulation of endonasal fluid exchange. PMID- 8231579 TI - Experimental reconstruction of anterior and circumferential defects of the cervical trachea. AB - Despite the variety of materials employable in the rehabilitation of the cervical trachea, long-term success is rarely achieved. Both gross and microscopic postoperative developments were studied on 45 dogs submitted to nine different types of reconstruction of anterior and circumferential defects. In the majority of cases, the grafts and prostheses used for repairs, enveloped by connective and inflammatory tissue, were either absorbed or expelled. Only pedicled autologous grafts survived rejection. Failure of tracheal replacements appears to be confined to tracheal and local phenomena. Such events impaired the incorporation of any type of graft or prosthesis. PMID- 8231580 TI - Indications for the AO plate with a myocutaneous flap instead of revascularized tissue transfer for mandibular reconstruction. AB - Our preferred technique for mandibular reconstruction uses a revascularized osseomyocutaneous flap. However, to reconstruct small lateral defects in dentulous patients, the AO mandibular reconstruction plate (MRP) is used. Its advantages include quick, reliable placement without a donor site or the need for advanced technical training. This review examines 16 patients who underwent primary mandibular reconstruction after ablative oncologic surgery using an AO MRP without a bone graft. The average follow-up was 32 months. Twelve patients (75%) had successful reconstructions, while 4 (25%) developed serious complications resulting in failure of the MRP. The most significant factor limiting rehabilitation was disease progression, affecting 12 (75%) of the 16 patients. In this subgroup of selected patients with small (< 6 cm) lateral mandibular defects, useful dentition, and advanced disease, or whose general medical condition precludes prolonged general anesthesia, primary mandibular reconstruction using an AO MRP and a myocutaneous flap without a bone graft is a viable reconstructive alternative. PMID- 8231581 TI - A treatment for primary ciliary dyskinesia: efficacy of functional endoscopic sinus surgery. AB - Primary ciliary dyskinesia (PCD) is an inherited disorder manifested in children as chronic otomastoiditis, recurrent pneumonia, and chronic sinusitis. The failure of the ciliary beat pattern to effectively function in the respiratory tract produces stasis of secretions with secondary inflammation, edema, and infection. The authors report three young children with PCD who presented with variable severities of symptoms. Each had the aforementioned respiratory tract problems. The child with the most severe symptomatology was treated with a variety of medical options, including long-term gamma globulin injections, but hospitalizations persisted twice per month because of severe sinopulmonary illness. All three of the children underwent functional endoscopic sinus surgery (FESS). In addition, two children received pressure equalization (PE) tubes. One child required a revision procedure. Their surgical outcomes are discussed. Follow-up of FESS in three children with this disorder shows a marked improvement in symptomatology with a decreased incidence of hospitalization and a somewhat decreased need for medical therapy. PMID- 8231582 TI - Partial cricoid resection with primary tracheal anastomosis for subglottic stenosis in infants and children. AB - Fifteen infants and children with a severe subglottic stenosis underwent a partial cricoid resection with primary tracheal anastomosis. The etiology of the stenosis was congenital in 3 cases and was acquired after prolonged intubation in 12. Fourteen patients were tracheotomy dependent at the time of surgery, and 13 (87%) of the 15 were classified grade III (7 cases) and IV (6 cases) according to Cotton. The resection included the cricoid only in 5 cases and the cricoid and a segment of trachea (1 to 4 rings) in 10 cases. Decannulation was achieved after a single open procedure in 14 (93%) of the cases. The authors encountered no lesion of the recurrent laryngeal nerves and no fatalities. There was, however, 1 restenosis. Twelve cases were decannulated within 6 months, most of them at 2 months. The postoperative voice is normal in 10 cases, and a slight dysphonia is present in 4 cases. Twelve patients show no exertional dyspnea, and 2 exhibit a slight stridor while practicing sports. Ten of 14 cases have been followed for more than 5 years, with the longest follow-up now being 14 years. In all cases, the laryngotracheal development is normal. The results of this small series compare favorably with those of laryngotracheoplasty procedures usually performed for subglottic stenosis in infants and children. In the future, partial cricoid resection with primary tracheal anastomosis should be considered as an important treatment option for severe subglottic stenoses in infants and children. PMID- 8231583 TI - Pitfalls in evaluation of photosensitizing agents: an example with Q-switch II dye. AB - Photosensitizing agents potentiating laser therapy should have limited toxicity, no mutagenicity, stable spectral characteristics, and acceptable solubility when administered in vivo. Q-Switch II dye (QII) has been shown by others to be an effective chromophore for photodynamic therapy at 1051 nm in fibroblast cell culture. The objective of this study was to determine the spectral stability of QII in biological media and then to localize QII after administration in vivo. Spectral evaluation was performed between 250 and 1100 nm. QII dissolved in dimethyl sulfoxide (DMSO) rapidly lost its spectral characteristics, including its 1051-nm peak, when contacting water, minimal essential medium, human serum, organ surfaces, and intracellular fluid. One minute following intramuscular (IM) injection of 0.1 mg QII in 0.2 mL of DMSO, the dose precipitated as a discrete mass which was excised and reconstituted in DMSO. A new spectral pattern was seen, with no absorption between 850 and 1100 nm. Following intravenous (i.v.), (IM), or intraperitoneal (IP) injection, QII was not detected in any organ. Q Switch II dye is not a suitable chromophore for in vivo photodynamic therapy at 1051 nm. Previous cell culture reports to the contrary did not account for the QII spectral change caused by biological media. Simple rapid assays are described to avoid this pitfall. PMID- 8231584 TI - Peritubal adenoidectomy. PMID- 8231585 TI - The subcutaneous pedicled extension flap: a new development in random flaps. PMID- 8231586 TI - Surgical anatomy of the glossopharyngeal nerve. PMID- 8231587 TI - Disinfection of laryngoscopes. PMID- 8231588 TI - Tympanostomy tube removal. PMID- 8231589 TI - Neurotologic skull base surgery for glomus tumors. Diagnosis for treatment planning and treatment options. PMID- 8231590 TI - Radiation therapy of glomus jugulare tumors. PMID- 8231591 TI - Basic surgical principles of neurotologic skull base surgery. PMID- 8231592 TI - The history of glomus tumors: a personal perspective. PMID- 8231593 TI - Rehabilitation of cranial nerve deficits after neurotologic skull base surgery. AB - Damage to any one of these nerves results in sufficient morbidity to warrant therapy; however, most patients will compensate for isolated loss of function. It is with multiple nerve injuries, as often seen in glomus surgery with the resection of cranial nerves IX, X, and XII that the full efforts of the rehabilitation team are called on. After vocal cord medialization and palatal adhesion, younger healthier patients will eventually resume adequate oral intake. However, the time it requires to return to a reasonably enjoyable diet often extends up to 1 year postoperatively. A few never attain the goal of enjoyable intake and continue to struggle to maintain adequate nutrition. The latter situation is the rule, not the exception, in the elderly population. Our experience over the years has led us to a more conservative treatment of glomus tumors in the elderly debilitated patient. PMID- 8231594 TI - Defect reconstruction following neurotologic skull base surgery. PMID- 8231595 TI - Surgery for glomus tumors: the Otology Group experience. PMID- 8231596 TI - The glomus tumor and its biology. AB - This section has attempted to detail the histopathologic, biologic, pathophysiologic, and epidemiologic characteristics of jugulotympanic paragangliomas. These features do not stand devoid of clinical significance, but rather lay the foundation for the formulation of logical diagnostic and therapeutic approaches to these fascinating tumors. PMID- 8231597 TI - Primum non nocere, the ethics of trials of leprosy chemotherapy. PMID- 8231598 TI - Management of plantar ulcers--theory or practice? PMID- 8231599 TI - Evaluation of the autoimmune response in leprosy. AB - Immunological responses to a panel of antigens were evaluated in 27 patients with lepromatous and 20 patients with tuberculoid leprosy and compared with 24 pulmonary tuberculosis patients, 25 systemic lupus erythematosus patients and 41 healthy blood donors. Some autoantibody specificities were extensively studied for the first time in mycobacterial infections. Striking immunoserological abnormalities were found in patients with lepromatous leprosy, particularly in those presenting with relapse. Inhibition assays were performed, providing a tool for further analysis of the binding range of specific anti-N.D.O. BSA antibodies and strengthening the suggestion of molecular mimicry reactions between cytoskeletal proteins, host stress proteins and Mycobacterium leprae antigens or stress proteins. A significant serological overlap between lepromatous leprosy and autoimmune diseases is indicated. PMID- 8231600 TI - Modulation of peripheral blood derived monocytes/macrophages from leprosy patients using 'tuftsin' for production of reactive oxygen intermediates. AB - Phagocytic cells respond to a variety of membrane stimulants by producing reactive oxygen intermediates (ROI), i.e. O2-, H2O2 and OH.metabolites. Plasma membrane activation is associated with superoxide generating NADPH oxidase, thereby causing the production of these toxic species. Stimulation of phagocytic cells also results in activation of purine catabolism, which directs the metabolic flux through xanthine oxidase to produce the superoxide anion. We previously observed that BL/LL macrophages (M phi) exhibited a premature inability to undergo tuftsin stimulated phagocytosis and microbicidal activity. The present study was undertaken to measure ROI levels in the absence and presence of 'tuftsin' pulsing as a function of in vitro culture age and also correlated these levels with adenosine deaminase (ADA) activity. The latter is known to be a contributor of O2- generation and is also involved in the maturation of the monocyte/macrophage system. The behaviour of normal and tuberculoid monocytes/macrophages were more or less the same, either in the presence or absence of tuftsin, i.e. they showed a progressive increase in ROI production until day 3, then tapered off in older cultures by day 7. In contrast, after day 1, the lepromatous macrophages were unable to undergo tuftsin mediated stimulation for the production of ROI and ADA activity. These findings indicate a defective M phi function in lepromatous patients towards tuftsin pulsing, thereby supporting our earlier observations. Thus BL/LL M phi behaved as if they were aged after 1 day of in vitro culture, which may account for an inability to handle Mycobacterium leprae for efficient killing. PMID- 8231601 TI - Reversal reaction in multibacillary leprosy patients following MDT with and without immunotherapy with a candidate for an antileprosy vaccine, Mycobacterium w. AB - Immunotherapy with a candidate for an antileprosy vaccine, Mycobacterium w, was given in addition to standard multidrug therapy (MDT) to 53 multibacillary lepromin negative patients belonging to BB, BL and LL types of leprosy (vaccine group). An equal control group received MDT and injections of micronized starch as placebo. Both the vaccine and placebo were administered intradermally every 3 months. The patients were evaluated at determined intervals by clinical, bacteriological and histopathological parameters and lepromin testing. Reactional episodes were analysed with reference to incidence, onset, frequency and severity during and after release from treatment (RFT). Incidence of reversal reaction (RR) was marginally higher in the vaccine group (22.6% vaccine group vs 15% control group). All cases with a history of downgrading type 1 reaction developed RR during therapy. Most episodes occurred within the 1st year of the commencement of therapy--50% developing within 3 months. Late reversal reaction (after RFT) were observed in 3.8% of cases in both groups, and 50% of the reactors in the control group and 33% in the vaccine group had repeated reactional episodes. Incidence of neuritis associated with RR as well as isolated neuritis was similar in both groups. PMID- 8231602 TI - Does the introduction of WHO-MDT influence trends in the incidence of leprosy?- the Malawian experience. AB - There has been an average annual decline in detection rates of all types of leprosy in Malawi of around 11.6% between 1977 and 1991. There was no obvious acceleration or slowing down of this decline following the introduction of WHO/MDT in 1983-84. Disability ratios stayed at the same level of about 11% during the 15 years covered by this paper suggesting that patients did not self report earlier after 1983-84 which might have masked an underlying accelerated decline in detection rates. Thus it is concluded that the influence of WHO/MDT on the pattern of leprosy over a period of time, in a country like Malawi, is so far not noticeably different from any influence dapsone monotherapy might have had. PMID- 8231603 TI - The effects of World Health Organization chemotherapy on imported leprosy in Auckland, New Zealand, 1983-90. AB - Between January 1983 and December 1990 in Auckland, New Zealand, 87 patients (28 paucibacillary disease (PBD) and 59 multibacillary disease (MBD)) commenced WHO multidrug therapy (MDT). All were immigrants from the Pacific Islands (65) or Asia (22). A total of 57 patients had already received non-WHO regimens, some continuously, but often intermittently, for many years; 30 patients received WHO MDT only. By December 1990, 50 had completed treatment, with 1 relapse and 1 late reaction, both in patients with PBD treated with WHO MDT only. There have been no relapses in those treated with WHO MDT after prior leprosy treatment. In those with MBD, type II leprosy reactions were less common (16%) in those treated only with WHO MDT than in those treated continuously before 1983 with older regimens (64%). Type I leprosy reactions occurred in about 20% of both these groups. The bacterial index fell faster in those who had had a prolonged prior treatment beginning WHO MDT than in those starting WHO MDT as their initial leprosy chemotherapy. Overall we found WHO MDT was well accepted and the compliance good, but 13 patients (15%) left Auckland before treatment was completed and 6 (7%) during follow up. PMID- 8231604 TI - Further observations on MDT blister-calendar packs in vertical leprosy eradication programmes--a multicentre study (phase II) AB - To improve operational efficiency as well as to improve patient compliance in leprosy programmes, DANIDA introduced blister-calendar packs (BCP) to deliver MDT in 4 MDT districts in India in 1987. An objective study (Phase II) involving 343 patients in a trial group (BCP group) and 253 patients in a control group (loose drug group) showed no significant difference in compliance rates for self administered dose between the 2 groups. Hence, while assessing the use of BCPs in leprosy programmes, other operational benefits like safe storage, easy transportation, easy drug accounting and safe preservation at home are to be considered. These aspects were followed up from Phase I of the study. PMID- 8231605 TI - Trigeminal neuralgia--a presenting feature of facial leprosy. AB - Trigeminal neuralgia is a well recognized clinical entity. However, it has not been reported to mimic leprosy or vice versa. Of the 3 cases reported here, 2 initially presented with neuralgic symptoms similar to that seen in trigeminal neuralgia and later developed borderline lesions on the face. The 3rd case demonstrated a tingling sensation along with firm and palpable supraorbital nerve (a branch of trigeminal nerve), and a very early skin lesion on the face pointed to the need to consider neuritic type leprosy before concluding the final diagnosis of a disease like trigeminal neuralgia which calls for a different therapeutic approach. PMID- 8231606 TI - Evolution of early lesions in leprosy. AB - We observed 29 patients presenting with vague peripheral neurological symptoms for 6 months or more. During this period, 16 developed clinical leprosy, 3 developed borderline tuberculoid leprosy and the other 13 developed neuritic leprosy. Of these 13 cases 11 subsequently developed skin lesions similar to those seen in indeterminate and in borderline tuberculoid leprosy. Based on the above observations, an attempt has been made to explain the evolution of early lesions of leprosy. PMID- 8231607 TI - Human immunodeficiency virus and leprosy--type 1 reactions, nerve damage and steroid therapy: 'a case report'. AB - In this study a 28-year-old female with both BL leprosy and HIV infections is discussed. Her clinical progress was followed until she completed MDT. During this period she developed recurrent reactional episodes, nerve damage and intercurrent illnesses--some of which might have been due to steroids. PMID- 8231608 TI - Squamous cell carcinoma in plantar ulcers in leprosy. A case control study. AB - The objective of this case-control study was to identify factors associated with the development of squamous cell carcinoma (SCC) in plantar ulcers of leprosy patients. We examined 2 matched groups consisting of leprosy patients with and without SCC in a plantar ulcer. No correlations were found between the development of SCC and race, profession, place of origin, duration of leprosy, the type and duration of leprosy chemotherapy, presence of bone involvement and type of ulcer care treatment given. The only statistically valid finding was that the duration of the ulcer was significantly lower in the group with malignant change. In this group there was an apparently higher use of pesticides, the difference being not of statistical significance. It is concluded that factors other than ulcer duration need to be looked for, in order to identify factors influencing malignant change in plantar ulcers of leprosy patients. PMID- 8231609 TI - PGL-I antibody in HIV infected patients. PMID- 8231610 TI - Side-effects of Isoprodian compared with WHO-MDT in rural Nepal. PMID- 8231611 TI - Comment: 'The allocation of leprosy patients into paucibacillary and multibacillary groups for multidrug therapy, taking into account the number of body areas affected by skin, or skin and nerve lesions'. PMID- 8231612 TI - Comment: 'Results of surgical procedures for the correction of foot-drop and of lagophthalmus due to leprosy'. PMID- 8231613 TI - [Treatment of gigantocellular tumors of the bone in our department]. AB - Thirty-four patients with giant cell tumor of bone were followed up at the Department of Orthopedics, University Hospital Firule, Split from 1960 to 1989. The age and sex distribution of patients and the location of tumors reported in this study are similar to those presented in the majority of large series. Thirty patients were treated with surgery and four with irradiation only. A significant correlation was observed between therapeutic procedures and recurrence of tumors. The incidence of local recurrence was 14.7%. The metastatic lesions in the lung were present in 26.5% of the patients. The most successful therapies for giant cell tumors are excision or resection "en bloc" and cancellous bone grafting. The authors recommend radical resection of tumors. PMID- 8231614 TI - [The value of exercise tests in the diagnosis of ischemic heart disease]. AB - New knowledges of contemporary cardiology have created possibilities for critically reevaluation of current criteria in exercise testing and have shown its new diagnostic and prognostic value. A multifactorial approach to exercise testing includes an analysis of ECG criteria and hemodynamic parameters obtained during exercise testing. This approach increases the value of stress testing in diagnosis, assessment of the severity and prognosis of coronary artery disease. The inclination toward arrhythmias, detection of asymptomatic ischemia and early detection of a dysfunction of the left ventricle have been emphasized. The indications for use and termination of exercise testing absolute and relative contraindications for exercise testing are critically discussed. The sensitivity of exercise testing is 60 to 80 percent and specificity level ranges between 85 to 90 percent. The incidence of serious complications is decreased about 50 percent by a careful screening of patients and appropriate safety level. PMID- 8231615 TI - [Histology of bone tissue and histomorphology--report on standardized terminology]. AB - The aim of this paper was to present and describe standardized terminology used in histologic and histomorphometric analysis of bone tissue. Bone tissue is characterized by specific activities which are the result of cell function throughout life. Histologic analysis of bone tissue specimen provides an insight in the features and quality of cellular activities. Histomorphometry is direct measurement and calculation of many parameters which permits quantification of characteristics and dynamics of particular bone tissue function. This method may be applied in diagnostics and monitoring of therapy effects in different metabolic bone disorders. PMID- 8231616 TI - [Arterial hypertension in the obese--aspects of etiopathogenesis]. AB - Many epidemiological and clinical studies show a strong association between arterial hypertension and obesity. The underlying pathophysiological mechanism is unknown. It is thought that the etiopathogenesis of obesity hypertension is exceptional and in that view hormonal, neural, volume and hemodynamic properties of obesity, as well as salt and/or caloric consumption, are outlined. In this article all these factors are discussed. According to the current hypothesis, hyperinsulinemia which is probably a physiologic adaptation to obesity, plays a key role in the pathogenesis of arterial hypertension. Insulin increases the reabsorption of sodium by means of an immediate effect on the kidney tubules. An increase of sodium in the body leads to hypervolemia and to the elevated blood pressure. Chronic hyperinsulinemia perhaps increases the blood pressure indirectly also by means of the central nervous system, namely, by stimulating the activity of the sympathetics. PMID- 8231617 TI - [Epidemiologic and clinical picture of anthrax in patients in Split 1956-1987]. PMID- 8231618 TI - [Familial aggregation of psoriasis]. AB - Eight hundred and eight patients with psoriasis, 490 males and 318 females, what was 64% of all the patients with psoriasis in the region of Karlovac on June 30, 1991, were included in the study. The family onset rate of the disease was 33.3% and psoriasis occurred most frequently among first-degree relatives. The distribution of psoriasis between brothers and sisters of our patients was 8.09%. If the parents were affected with the disease, the percentage of brothers and sisters, with psoriasis increased proportionally. The risk of psoriasis among fraternal relatives is 25.7% according to Weinberg. These findings suggest that psoriasis is determined by genetic factors. The age of onset of psoriasis was analyzed with respect to familial occurrence. It was found that patients with psoriatic relatives had a 10-year earlier onset of the disease, particularly females, compared to those with negative history. By 25 years of age, 58% of the patients with positive and 35% with negative history were affected with the disease. These observations are consistent with the reports of some authors that an earlier onset of the disease announces familial frequency. PMID- 8231619 TI - [The effect of atmospheric conditions on the occurrence of peptic ulcer hemorrhage]. AB - The influence of atmospheric factors on the frequency of bleeding from the peptic ulcer was studied within the period from April 1, 1984, to March 31, 1989, consequently through 1826 days. The average daily atmospheric pressure, the average daily temperature and the relative humidity have been examined. There were 1102 cases of bleeding peptic ulcer, 537 bleeding gastric ulcers and 565 bleeding duodenal ulcers. During the study period there were 454 days with bleeding form ventricular ulcer and 465 days with bleeding from duodenal ulcer. There was 793 days with bleeding form either lesion. The discriminatory analysis demonstrated that the atmospheric pressure is the variable that discriminates the days with bleeding and the days prior to bleeding from the days without bleeding. The relative humidity occurs as the relevant discriminatory variable in the days prior to bleeding for the duodenal ulcer group and for the entire group. The centroids of the discriminatory function demonstrate that the days with ulcer bleeding are characterized by the fall of atmospheric pressure. The factor analysis of meterological variables clearly shows the correlation of the atmospheric pressure and the bleeding regardless to the localisation of bleeding ulcer, where the greatest number of bleedings is correlated with lower atmospheric pressure. We conclude that the incidence of bleeding form the peptic ulcer of the stomach and duodenum correlates in great measure with low atmospheric pressure in the days prior to bleeding and in the days of bleeding, as well as with fall of atmospheric pressure in the days of bleeding with respect to previous day.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231620 TI - [The effect of chronic kidney disease on the course and outcome of pregnancy]. AB - In order to estimate the effect of renal disease on the pregnancy, and the effect of pregnancy on the natural course of renal disease, the course and outcome of the 37 pregnancies was analyzed in gravidas with chronic renal disease. The women were cared for between 1978 and 1990 at the Department of Obstetrics, School of Medicine University of Zagreb. Analysis of the results in this article is retrospective. Thirty-six pregnancies finished by vaginal or cesarean delivery or by abortion, while one pregnancy was ectopic and ended by laparotomy during the 20th week. There were 31 live births from 36 fetuses (86.1%), 5 stillbirths (13.19%), of which 2 pregnancies ended in fetal death (abortion). In addition, there were 6 instances of neonatal death (8.3%). Twenty-five percent of pregnancies finished before 37 weeks of gestation. In 6 of 34 (17.6%) deliveries amniotic fluid was meconium stained, and 5 of 31 (16.1%) infants were born hypoxic. There were 29 percent of growth retarded liveborn fetuses. Perinatal mortality was 167/1000. Renal insufficiency was noted in 12 of 37 (32.4%) pregnancies, 54% of pregnant women had hypertension, 8.1% hyperkalemia, 78.4% were anemic, 27% had significant bacteriuria and 21.6% overt pyelonephritis. There was one case of acute deterioration in renal function that required hemodialysis, and one case of preeclampsia. Renal insufficiency or hypertension reduce drastically the chances for a successful outcome of pregnancy in gravidas with kidney disorders. However renal insufficiency in the presence of hypertension, carries even poorer prognosis, with perinatal mortality of 428/1000. PMID- 8231621 TI - [Growth and development in children with congenital heart defects]. AB - Growth and development have been analyzed in 222 children with congenital heart disease. Results were compared to a group of 50 control children. There was no significant difference in age and sex between the groups (p < 0.05). Thirty-three children (12.16%) had cynotic congenital heart disease (tetralogy of Fallot and transposition of the great arteries) and 189 (87.84%) acyanotic congenital heart disease (ventricular septal defect, atrial septal defect, patent ductus arteriosus and aortic and pulmonary stenosis). A statistically significant growth retardation was noted in all children with cardiac disease as compared to the control group, and it was evaluated by percentile values of body weight and height (p < 0.01). Weight retardation was more marked than retardation in body height (p < 0.01). Growth retardation was more significant in the cyanotic children that in those with acynotic heart disease (p < 0.01). Among the children with left-to-right intracardial shunt, growth retardation was found to increase proportionally with the size of the shunt and was most significant in patients with large left-to-right shunt (QP/QS > 1.80). No statistically significant difference in growth retardation was established among the children with acyanotic heart disease (p > 0.01). PMID- 8231622 TI - [Clinico-laboratory parameters of bacteremia and sepsis caused by Escherichia coli]. AB - Clinical and laboratory features were analyzed in 277 patients with 279 episodes of E. coli bacteremia, treated at the University Hospital of Infectious Diseases "Dr Fran Mihaljevic"c, Zagreb between 1980 and 1989. Patients ranged in age from 4 days to 91 years. Thirty-two per cent of patients were male and 68% were female. The majority of patients had underlying disease which was considered not to be fatal within the next five years (57%) or they were completely healthy (34%). Four per cent of bacteremias were polymicrobic and 13% were considered nosocomial in origin. The clinical source of bacteremia was known in 84%. In community-acquired bacteremia, the urinary tract was the most frequent site of origin (78%) and in nosocomial-acquired bacteremia the source could not be identified (54%). Septic shock occurred in 24% of the patients and coagulation abnormalities were observed in 41% of the patients. Fatal outcome was registered in 10% of the patients. Mortality was higher among males (16%:7%) up to 2 months of age (29%) and above 60 years (15%), and in those with nosocomial-acquired bacteremia (49%), polymicrobial (50%) and recurrent bacteremia (50%), as well as in patients with severe underlying disease (29-75%). Mortality was lower in patients who were given an early appropriate antimicrobial therapy. The advantage of combinations of antimicrobial therapies over monotherapy could not be demonstrated among patients in each category of severity of underlying host disease. PMID- 8231623 TI - [Prevention of hepatitis B virus infection in individuals at high risk in the Cakovec Medical Center]. AB - The results of hepatitis B vaccination (H-B-VAX, 1 ml = 20 micrograms HBsAg, scheme 0, 1, 6) in nine susceptible health care workers of hemodialysis unit are described. Seroconversion was achieved in 100% of recipients. The mean titer of anti-HBs was 11,691 mIU/ml at three years, 5,879 mIU/ml at four years and 2,770 mIU/ml at five years after the beginning of vaccination. H-B-VAX vaccine (first dose 40 micrograms HBsAg, second and third = 20 micrograms) was administered in three patients on hemodialysis (scheme 0, 1, 2). In all of these patients, seroconversion occurred (titers of anti-HBs from 23 to 104 mIU/ml). Of the 55 workers of dental units, nine (16.4%) showed signs of previous HBV infection. Twenty were vaccinated with H-B-VAX vaccine (scheme 0, 1, 2) and of these patients one (5%) was nonresponder, two (10%) poor responders, 13 (65%) good responders and four (20%) excellent responders, with the mean anti-HBs titer of 1,540 mIU/ml. Seven dental workers received recombinant vaccine also using a scheme 0, 1, 2. Four (57%) of these workers were poor responders, and three (43%) good responders, with the mean anti-Hbs titer of 168 mIU/ml. All of our vaccinated subjects had no serious side effects except local soreness at the injection site. Thus, vaccination against hepatitis B in health care personnel and other high-risk patients may control efficiently the spread of HBV infection. PMID- 8231624 TI - [Pulmonary changes in patients with type I neurofibromatosis and tuberous sclerosis]. AB - Two patients with pulmonary affections in the course of neurofibromatosis and tuberous sclerosis are presented. We wanted to draw attention to the possibility of occurrence of diffuse interstitial pulmonary changes during congenital neurocutaneous syndromes, of which no data are reported in our literature. The affection of the lungs in 10-20% of the patients with neurofibromatosis and 1% with tuberous sclerosis is estimated, commonly in women. The most frequent changes in neurofibromatosis are fibrosing alveolitis and interstitial fibrosis, and in tuberous sclerosis cystic lung changes and lymphangiomyomatosis, what agrees with our findings. It is important to recognize such patients in order not to perform unnecessary diagnostic procedures, and not to draw wrong conclusions, thus being able to avoid an unnecessary therapy, ineffective in such cases. The patients should be followed up, and in the case of complaints symptomatic therapy has to be administered. PMID- 8231625 TI - [Surgical treatment of congenital obstruction of the left ventricular outflow tract]. AB - This report presents the classification and all types of left ventricular outflow tract obstructions. The possibilities of operative therapies are surveyed as well. Results of surgical treatment in 34 patients with obstruction to left ventricular outflow are shown. The majority of patients underwent operation under extracorporeal circulation (84.4%), while the rest were operated by means of the inflow occlusion technique (14.7%). The obtained results were compared with those from the literature. The importance of echocardiographic evaluation of location of the left ventricular outflow tract obstruction and the appropriate choice of a surgical technique according to the patient's age are emphasized. PMID- 8231626 TI - Mouse sperm-egg interaction in vitro in the presence of neem oil. AB - In vitro evidence is presented showing toxicity of neem oil on sperm-egg interaction in mouse. Cumulus oophorus-enclosed ova, inseminated with capacitated spermatozoa, were cultured in 1 ml of in vitro fertilization (IVF) medium and overlayered by 1 ml of different concentrations of neem oil (1, 5, 10, 25, 50 and 100%) for IVF duration of 4h. At the end of incubation, ova were allowed to grow in neem oil-free culture medium and assessed for fertilization, first cleavage (2 cell formation) and blastocyst formation in vitro at 4-14h, 24h and 108h post insemination respectively. The study showed that the presence of neem oil at concentrations of 10, 25 and 50% caused inhibition of IVF in a dose-dependent manner. The toxic effect of exposure of 25 and 50% neem oil was further carried over to the first cleavage of the resulting fertilized ova and the toxic effect of 5, 10, 25 and 50% was carried over to the blastocyst formation from the resulting fertilized ova when grown in neem-oil free culture medium. A total of 94.1% inhibition of 2-cell formation and 100% inhibition of blastocyst formation from the inseminated ova was observed in 50 and 25% neem oil-treated groups respectively. Neem oil at 100% concentration caused 100% degeneration of ova at 1h of sperm-ova coculture. The study showed a direct toxic effect of neem oil on sperm-egg interaction in vitro and encourages research investigations of this herbal product as a pre-coital contraceptive. PMID- 8231627 TI - Differential cerebrovascular responsiveness in spontaneously hypertensive rats following antihypertensive treatment with clonidine and verapamil. AB - Numerous studies have been reported examining the effects of antihypertensive treatment on peripheral vascular responsiveness in spontaneously hypertensive rats (SHR). This study was conducted to determine the effects of chronic treatment with 2 antihypertensive agents on cerebrovascular responsiveness in male SHR and Wistar-Kyoto (WKY) rats. SHR and WKY (3-4 weeks old) received either placebo, clonidine (CLON, 10 mg pellet) or verapamil (VER, 5 mg pellet). Vascular reactivity studies on the basilar artery, using standard smooth muscle bath techniques, were conducted following 6 weeks of treatment. Both CLON and VER significantly attenuated the rise in blood pressure in SHR. Basilar artery responsiveness to KCl, serotonin (5-HT), and calcium were significantly increased whereas responses to acetylcholine (ACH), isoproterenol (ISO) and sodium nitroprusside (SNP) were significantly reduced in SHR compared to WKY. CLON had no effect on basilar artery responsiveness to either the contractile or relaxation agents in SHR. However, although responses to KCl, 5-HT and calcium were not affected by VER in SHR, VER significantly increased the responses to ACH, ISO and SNP. Neither CLON nor VER treatment affected basilar artery responsiveness to any of the agents in WKY. These data demonstrate that, even though CLON and VER have similar antihypertensive effects, differential effects of the 2 agents on cerebrovascular responsiveness in the SHR are apparent. This would suggest that the vascular effects of VER and CLON are dependent upon the mechanism of action of the agents and not simply due to prevention of the elevation in blood pressure. PMID- 8231628 TI - Pyrogenic doses of intracerebroventricular interleukin-1 did not induce analgesia in the rat hot-plate or cold-water tail-flick tests. AB - There are a few reports in the literature that cytokines can induce analgesia (5, 6, 18). The present study sought to characterize the analgesic effects of intracerebroventricularly (icv) administered interleukin-1 (IL-1) and interferon alpha (IFN-alpha) in rats. In the cold-water tail-flick test (CWT), latency to tail withdrawal from a -3 degrees C liquid was timed; in the hot-plate test (HP), latency to a rear paw lick or a jump from a 55 degrees C surface was measured. In some experiments, core body temperature was also monitored with a rectal thermistor. In the CWT, human recombinant (hr) IFN-alpha induced a small, statistically significant effect at just one dose (15,000 U icv), but no dose of hr-IL-1 alpha (250-1000 U icv) or hr-IL-1 beta (125-2000 U icv) induced a significant effect at any time point. On the other hand, dose-related increases in body temperature were observed after icv injection of both IL-1 alpha and IL-1 beta. The largest hyperthermic effect was a 1.7 (+/- 0.15) degrees C rise 120 min after administration of 1000 U IL-1 beta. In a second analgesic assay, the HP, IL 1 beta was ineffective as well. Since IL-1 alone did not induce analgesia, we tested its capacity to potentiate morphine analgesia. Morphine (5.0 and 10 micrograms, icv) induced analgesia in the CWT (32.7 and 61.8% maximum analgesia, respectively); however, there was no significant effect of IL-1 beta on morphine induced analgesia. In summary, we failed to find an analgesic effect of IL-1, alone or in combination with morphine, at doses which clearly had a physiological effect; this is in contrast to the reports cited above. PMID- 8231629 TI - Deficiency of holo-, but not apo-, ceruloplasmin in genetically copper intoxicated LEC mutant rat. AB - Long-Evans Cinnamon(LEC) mutant rats exhibited less than 5% of normal levels of serum ceruloplasmin oxidase activity, but immunoblot analysis showed normal levels of immunologically detectable ceruloplasmin protein in sera from the mutant rats. Immunostaining of cryosections from the liver tissues with anti ceruloplasmin antibody showed no significant difference between normal and LEC rats. Results from pulse labeling of ceruloplasmin for 3 hours with [35S]methionine in primary hepatocyte culture, followed by immunoprecipitation, SDS-PAGE and fluorography, showed only minor changes in ceruloplasmin protein synthesis and secretion. These results suggest that the mutation(s) does not affect ceruloplasmin gene expression, but results in a failure in the mechanism whereby copper is incorporated into newly synthesized apoceruloplasmin to produce oxidase active holoform. PMID- 8231630 TI - Phencyclidine exposure alters in vitro cellular immune response parameters associated with host defense. AB - Phencyclidine hydrochloride (PCP) was tested for its ability to alter a variety of immune effector and regulatory functions in vitro. B6C3F1 murine splenic lymphocytes or elicited peritoneal macrophages were cultured in vitro with medium only or medium containing 10(-10)-10(-4) M PCP. Macrophages cultured with or without PCP were stimulated with lipopolysaccharide, and production of interleukin 6 (IL-6) and tumor necrosis factor (TNF) was assessed by bioassay. Cytotoxic T-cell effector function was determined following 5-day lymphocyte co culture with tumor stimulator cells in the presence of PCP. In addition, the ability of T-lymphocytes to produce specific immunoregulatory cytokines IL-2 and IL-4 in the presence of PCP was quantitated by bioassay. B-lymphocyte function was determined by quantitating lymphocyte proliferation following stimulation with anti-IgM antibody and murine IL-4. Natural immunity was assessed by culturing lymphocytes with or without PCP for 24 h, then quantitating basal and IL-2 augmented natural killer (NK) cell activity. In the absence of effects on cell viability, significant suppression of IL-2 production by T-cells was noted at pharmacologically relevant PCP concentrations (1 microM). In vitro concentrations of 10 microM suppressed the generation of specifically sensitized cytotoxic T-cells. In addition, PCP significantly suppressed both IL-2-augmented NK function as well as B-lymphocyte proliferation. By comparison, macrophage IL-6 production was not affected by any concentration of PCP examined in this study. PMID- 8231631 TI - In vitro effects of psychoactive and non-psychoactive cannabinoids on immature rat Sertoli cell function. AB - We have examined the effects of psychoactive and non-psychoactive cannabinoids on isolated immature rat Sertoli cells cultured in either serum-free or serum containing media. Lactate accumulation by Sertoli cells in serum-free control cultures was 10 fold greater than the values obtained in control cultures exposed to serum. Under either culture condition, 3.1 micrograms/ml of delta-9 tetrahydrocannabinol (THC) or cannabinol (CBN) stimulated lactate secretion above control levels. Cannabidiol (CBD) stimulated lactate secretion in serum containing but not in serum-free media. Using serum-free culture conditions, we next studied the in vitro effects of combinations of THC and either epinephrine or FSH on lactate and transferrin secretion by immature rat Sertoli cells. Co incubation of 0.1 microM epinephrine with 0.8 or 3.1 microgram/ml THC significantly stimulated lactate secretion when compared to epinephrine or THC alone, while only the high THC dose increased transferrin secretion. Moreover, co incubation of FSH (1 micrograms/ml) with THC (0.8 or 3.1 micrograms/ml), significantly stimulated both lactate and transferrin production by immature rat Sertoli cells. These results add to the growing evidence that cannabinoids can exert direct effects on Sertoli cell function and modulate their responses to physiological stimuli. PMID- 8231632 TI - Glucagon modulates superoxide generation in human polymorphonuclear leucocytes. AB - It has been found that leucocytes possess receptor sites for glucagon and glucagon was shown to increase during bacterial infection. To verify the interconnection between glucagon, leucocytes and bacterial infection we studied the effect of glucagon on superoxide generation and second messenger transduction in PMNs. We found that glucagon could not stimulate chemiluminescence by itself but it could enhance FMLP- but not PMA-induced chemiluminescence in a concentration (50-800 pg/ml) dependent manner. However, after incubation of PMNs with 10 microM of ST-638 (a tyrosine kinase inhibitor) the enhancement effect converted into inhibitory effect. We also found that glucagon treatment of PMNs increased both IP3 and cyclic AMP levels as second messengers. ST-638 greatly attenuated the IP3 increment in the glucagon-treated FMLP-stimulated PMNs. From these results we can conclude that glucagon could enhance superoxide generation from FMLP-stimulated PMNs by elevating IP3. Inhibition of IP3 increment by tyrosine kinase blockade uncover the inhibitory effect of the increasing cyclic AMP on superoxide production. PMID- 8231633 TI - Effects of calcium channel blockers on bupivacaine-induced toxicity. AB - The purpose of this study was to investigate the influence of calcium channel blockers on bupivacaine-induced acute toxicity. For each of the three tested calcium channel blockers (diltiazem, verapamil and bepridil) 6 groups of mice were treated by two different doses, i.e. 2 and 10 mg/kg/i.p., or an equal volume of saline for the control group (n = 20); 15 minutes later, all the animals were injected with a single 50 mg/kg/i.p. dose of bupivacaine. The convulsant activity, the time of latency to convulse and the mortality rate were assessed in each group. The local anesthetic-induced mortality was significantly increased by the three different calcium channel blockers. The convulsant activity of bupivacaine was not significantly modified but calcium channel blockers decreased the time of latency to obtain bupivacaine-induced convulsions; this effect was less pronounced with bepridil. PMID- 8231634 TI - Plasma catecholamine, renin activity, and ACTH responses to the serotonin receptor agonist DOI in juvenile spontaneously hypertensive rats. AB - Mean arterial blood pressure (MAP), heart rate (HR) and arterial plasma levels of corticotropin (ACTH), renin activity (PRA) and catechols [norepinephrine (NE), epinephrine (EPI), and the intraneuronal NE metabolite dihydroxyphenylglycol (DHPG)] at baseline and in response to the serotonin-1C/2 (5-HT1C/5-HT2) agonist 1-(2,5-dimethoxy-4-iodophenyl)2-aminopropane (DOI, 1.0 mg/kg i.a.) in conscious, freely-moving, juvenile (4 week old) spontaneously hypertensive rats (SHR's) and age-matched Wistar-Kyoto (WKY) normotensive control rats were measured simultaneously. Baseline levels of MAP, NE, DHPG, and EPI all were significantly higher in the SHR's. There was a similar trend for PRA, but ACTH did not differ between the two strains. DOI produced marked increases in levels of MAP, ACTH, EPI, and also PRA but did not affect NE or DHPG concentrations. HR decreased only in the WKY group after administration of DOI. The magnitudes of the EPI and ACTH responses did not differ between the rat strains. Responses of MAP and PRA were significantly larger in SHR's. These results suggest that there is a selective hyperresponsiveness of PRA and blood pressure to 5-HT2 receptor stimulation parallel to a deficient baroreceptor reflex in juvenile SHR's. PMID- 8231635 TI - Detection and evaluation of persisting stress-induced hormonal disturbances by a post stress provocation test in humans. AB - Eighteen healthy army officers were subjected after prolonged rest to exhaustive ergometric work for about 15 minutes. Before and afterwards blood was taken from the cannulated antecubital vein for determination of free and sulfoconjugated catecholamines, cortisol, glucose, and white blood cell count. One week later, the same procedure was repeated with the same subjects with the difference that the probands underwent about 2.5 hours of difficult mountain climbing and a subsequent rest of 1.5 hours before ergometry. The most important results were: 1) total and bound fractions of catecholamines showed some significant differences between the first and second ergometry due to the previous mountain climbing stress; 2) serum cortisol did not increase after the first ergometry but did so significantly after the second ergometry due to the previous stress; low cortisol is not always indicative of the absence of stress; 3) the absolute number of white blood cells increased in both situations, correlated significantly with the severity of the stress and the individual increases were more person than situation specific and; 4) blood glucose remained unaffected in both situations. We conclude that a previous stress experience can affect a second stress response and that such a post stress provocation test can uncover persistent hormonal alterations. This procedure may be useful for the evaluation of inaccessible stress situations from subsequent stress measures. PMID- 8231636 TI - Suppressive effect of soybean milk protein on experimentally induced skin tumor in mice. AB - We studied the effect of soybean milk protein (SMP) in a two-stage carcinogenesis experiment on mouse skin. Mice were given soybean protein isolate (SPI) diet or SPI diet supplemented with SMP. After 4 weeks on the diets, the mice were shaved and a tumor initiator was applied. A tumor promotor was then applied twice a week on the same area of the skin throughout the experiment. After 20 weeks on the treatment, the percentage of tumor-bearing mice and the volume of tumor tended to be lower in the mice on the SMP diet than those on the SPI diet. The number of tumor was also significantly lower in the former group as compared to the latter group. There was no difference in growth between mice of the SPI and SMP groups. The results indicate the safety and the anti-carcinogenic effect of SMP in mice. PMID- 8231637 TI - Determination of plasma volume with indocyanine green in man. AB - We investigated the feasibility of using indocyanine green (ICG) for plasma volume (PV) determination in man. Duplicate PV measurements were carried out in 23 healthy subjects to test repeatability. ICG (0.25 mg/kg) was injected intravenously into one arm and venous blood was withdrawn from the opposite arm. Optical density of plasma samples from minute 3 to 9 was measured in a densitometer. ICG concentration at injection time was determined by monoexponential extrapolation. The mean (SD) difference (MD) was -23 ml (183) or 0.6% (5.7%). Linear regression revealed PV2 = 0.92.PV1 + 226 (r = 0.92). The PV values corresponded well with data from other studies. In 26 surgical patients PV was determined using two methods: 1) the same as in healthy subjects and 2) using a modification of this method in whole blood (PVB). For PVB measurement blood was drawn through a cuvette-densitometer from an arterial line. Calculations were the same as in PV determination except for the use of hematocrit to achieve plasma concentrations of ICG from whole blood. In patients MD were -53 ml (144) or -1.3% (4.3) for PV and -19 ml (161) or -0.3% (5.1) for PVB. Comparing PVB and PV revealed MD = -113 ml (149) or -3.3% (4.2). The whole blood method is easier to perform and reduces blood waste to almost zero. In conclusion, ICG is a suitable tracer for PV determination. PMID- 8231638 TI - Measurement of prolactin release and cytosolic calcium in estradiol-primed lactotrophs. AB - We have developed a perifusion system that can measure both changes of cytosolic free calcium concentration [Ca2+]i and prolactin release simultaneously from cultured lactotrophs. This model incorporated a commonly-used perifusion system to a spectrofluorometer. Indo-1 loaded cells were injected into Sephadex G-150 matrix in the cuvette at a site where the emitting light of the fluorometer projects. During perifusion periods, the perifusate was collected in a fraction collector, while optical density of the emitting light at 405 nm was recorded. The [Ca2+]i was calculated based on an ionomycin and Mn2+ quenching technique. As expected, TRH (1 mumol/l) stimulated prolactin release from cultured lactotrophs in this system. We further observed that prolactin releases as induced by TRH and ionomycin were not proportional with changes of the [Ca2+]i, suggesting that changes of [Ca2+]i is not the sole final pathway of intracellular transduction systems for prolactin release. PMID- 8231639 TI - Reduced sensitivity of cultured skin fibroblasts from familial hypercholesterolemic patients to glucocorticoids. AB - Cultured human skin fibroblasts, obtained from patients with homozygous familial hypercholesterolemia (FH), exhibited cholesterol synthesis that was 2.5-3-fold more intensive than in cells from healthy donors (normal cells). The study of the glucocorticoid [3H]-dexamethasone specific binding (glucocorticoid receptors, GcR) to the cells showed reduced number of binding sites (8 x 10(3) vs 120 x 10(3) binding sites/cell) and reduced Kd value (5.6 +/- 0.7 vs 10.1 +/- 2.1 nM) in FH-fibroblasts when compared with normal fibroblasts. The inhibition by 1 x 10(-8) and 1 x 10(-6) M dexamethasone of cholesterol synthesis was less marked in FH-cells (29.1 +/- 2.8 and 35.9 +/- 1.2%) as compared with normal cells (53.3 +/- 1.0% and 60.4 +/- 9.6, respectively). This findings indicate that the sensitivity of FH-fibroblasts to glucocorticoids is reduced. They also suggest that reduced number of GcR found in FH-fibroblasts reflects the cellular adaptation to cholesterol deficiency resulted from the absence of LDL-receptors. PMID- 8231640 TI - Contrasting action of antiestrogen (ICI-182780) for preventing initiation of embryo implantation by estradiol or epidermal growth factor (EGF). AB - The pure estrogen antagonist ICI-182780, at doses above 50 micrograms/kg, effectively inhibited the initiation of embryo implantation in rats when administered on day 4 of pregnancy (day 1 = sperm positive). The same dose inhibited the implantation initiating effect of intravenous 25 ng of estradiol-17 beta in delayed implanting progesterone-primed hypophysectomized rats. In contrast, the anti-estrogen at a dose of 1 mg/kg was ineffective at inhibiting the initiation of implantation induced by intrauterine plus intravenous administration of murine epidermal growth factor to delayed implanting rats. The growth factor also initiated implantation of blastocysts transferred from donor animals injected with the anti-estrogen to progesterone-primed hypophysectomized recipients. The results clearly demonstrate that the implantation initiating effect of the growth factor is not inhibited by a pure estrogen antagonist, and therefore this estrogenic function does not appear to require action initiated by the classical estrogen receptor. PMID- 8231641 TI - Effect of endothelin receptor antagonist, BQ-123, on Ca2+ signaling in cultured rat mesangial cells. AB - Endothelin (ET)-1 causes mesangial cell contraction and proliferation. The present study was designed to evaluate the functional ET receptor subtype in cultured rat mesangial cells by measuring intracellular Ca2+ signaling with the use of a newly synthesized ET receptor type A selective antagonist-BQ123. The ET 1, ET-2 and ET-3 increased intracellular Ca2+ level in a dose-dependent manner. BQ-123 suppressed intracellular Ca2+ elevation in response to ET-1 in a dose dependent manner with its half maximal inhibition value of 28 nM. BQ-123 (10(-6) M) did not affect ET-3 (10(-7) M)-induced Ca2+ response. The peak Ca2+ levels after addition of ET-3 without BQ-123 were similar to those in response to ET-1, ET-2 and ET-3 in the presence of BQ-123. Northern blot analysis showed that cultured mesangial cells expressed both ETA and ETB receptor mRNA. These results suggest that cultured mesangial cells have, at least, two types of functional receptors, one of which is an ETA receptor. PMID- 8231642 TI - 5-Methoxypsoralen: effect on the noradrenaline circadian rhythm of sleep-deprived subjects. AB - 5-methoxypsoralen (5-MOP), a serotonin analogue, stimulates the secretion of melatonin, which plays an important role in circadian rhythm regulation. Melatonin production is essentially controlled by noradrenaline (NA). To investigate the effect of 5-MOP on the 24 hr NA rhythm, hourly plasma NA concentrations measured over 24 hr in 7 healthy young subjects who took 40 mg 5 MOP orally at 21:00 hr the evening before were compared with values obtained in drug-free subjects. All subjects were sleep-deprived and under conditions in which sympathetic nervous system activation was repeated every hour over 24 hr. In both series of patients, a significant difference was observed between mean morning values and mean night values, reflecting persistence of a circadian rhythm. In subjects given 5-MOP, a significant difference was also observed between mean afternoon values and mean night values. However, MANOVA analysis failed to find any difference between the two series of subjects. Acute administration of 5-MOP thus had no significant effect on the NA circadian rhythm under our study conditions. PMID- 8231643 TI - Ketamine inhibition of cytoplasmic calcium signalling in rat pheochromocytoma (PC 12) cells. AB - This study examines the mechanism of action of ketamine, a dissociative anesthetic, with a specific focus on its ability to inhibit changes in the concentration of intracellular free calcium, [Ca2+]i, in PC-12 cells. The resting [Ca2+]i as measured with the fluorescent probe Fura-2 AM in control cells is 184.8 +/- 8.6 nM (mean +/- SEM, n = 15). Changes in [Ca2+]i via influx through voltage-gated calcium channels after membrane depolarization with potassium chloride were monitored in the absence and presence of various concentrations of ketamine. Potassium-depolarization caused a dose-dependent rapid increase in [Ca2+]i, averaging 62 +/- 5%, 33 +/- 2% and 18 +/- 3% (n = 10 each) above control levels for 70 mM, 50 mM and 35 mM KCl, respectively. Ketamine, in the dosage range studied (5-500 microM), inhibited the increase in [Ca2+]i stimulated by potassium-depolarization in a dose-dependent manner. The computer-fitted dose response curve of the pooled data yielded a half maximal suppression concentration, ED50, of 33 microM. In conclusion, this study demonstrates that ketamine inhibits Ca2+ influx through voltage-gated Ca2+ channels in PC-12 cells at clinically relevant doses, and may play a role in ketamine's action as a general anesthetic agent. PMID- 8231644 TI - Effects of Ca2+ channel blockers on physical dependence on diazepam in mice. AB - The effects of Ca2+ channel blockers on the development of physical dependence on diazepam were examined in mice. Co-administration of flunarizine (T-type Ca2+ channel sensitive blocker), but not of either nifedipine or diltiazem (L-type Ca2+ channel sensitive blockers), with diazepam significantly suppressed the hypersensitivity to FG 7142 following chronic treatment with diazepam. The hypersensitivity to FG 7142 may reflect benzodiazepine withdrawal convulsions. These results suggest that flunarizine, but not nifedipine or diltiazem, may suppress the development of physical dependence on diazepam, and that T-type Ca2+ channels in the brain, rather than L-type Ca2+ channels, may be involved in the development of physical dependence on diazepam. PMID- 8231645 TI - Decreased sensitivity of aorta from hypertensive rats to vasorelaxation by tyrphostin. AB - The role of protein tyrosine kinases (PTKs) in vascular smooth muscle (VSM) contraction was examined in spontaneously hypertensive rats (SHRs). Aorta from SHRs was hyperresponsive to PTK-mediated contraction relative to normotensive Wistar-Kyoto rats (WKYs). Aorta from SHR was also hyporesponsive to vasorelaxation by tyrphostin, a selective inhibitor of PTKs. Further, we found alterations in PTK activity in aorta from SHRs. PDGF stimulated PTK activity to a greater extent in the SHR. Tyrphostin inhibited PDGF-induced PTK stimulation in both strains, however, activity returned to basal levels in the WKY only. The results suggest that PTKs may be involved in VSM contraction and in the development of hypertension. PMID- 8231646 TI - The effect of adrenomedullin on the isolated heart. AB - A novel peptide found in human blood, adrenomedullin (ADM), has been shown to have systemic vasodepressor activity in the rat. However, the direct effects of ADM on cardiac function are unknown. Results of the present study demonstrate that ADM13-52 possesses marked systemic vasodepressor activity in the anesthetized rat. Although ADM13-52 modestly decreased peak systolic pressure (PSP) indicating mild negative inotropic activity, the present data suggest that bolus administration of ADM decreases systemic arterial pressure by dilating the systemic vasculature. The present data also suggest that only a portion of the ADM molecule is necessary to produce systemic vasodilation. PMID- 8231647 TI - Gastrointestinal effects of 5-hydroxytryptamine and related drugs. AB - This paper deals with the effects of 5-hydroxytryptamine (5-HT; serotonin) and related drugs on the gastrointestinal tract (GIT). The nomenclature and classification of 5-HT receptors, as well as their putative role in the GIT are updated in this review. Besides its effects on the cardiovascular system, which have been extensively described, several lines of evidence suggest a role for 5 HT in regulating gastrointestinal functions. 5-HT is present in the gastrointestinal tissues, and can elicit contraction or relaxation by activation of a wide variety of mechanisms and receptors. At least four main types of receptors (5-HT1, 5-HT2, 5-HT3 and 5-HT4) have been described and all the four types seem to influence the GIT. In this respect, the 5-HT2, and in some cases the 5-HT1 receptors, appear to be present on the gastrointestinal smooth muscle, while 5-HT3 and 5-HT4 are mainly neuronal. PMID- 8231648 TI - The behavioral and biochemical effects of thioperamide, a histamine H3-receptor antagonist, in a light/dark test measuring anxiety in mice. AB - We investigated the effects of thioperamide, a histamine H3-receptor antagonist, in a light/dark test measuring anxiety in mice. Thioperamide (20 mg/kg) slightly affected the locomotion and time spent in a light zone, and shuttle crossing. However, the decreases of these parameters were significant only when the animals were pretreated with zolantidine, a histamine H2-receptor antagonist. Moreover, the decreased parameters induced by the combination of thioperamide and zolantidine were reversed by pretreatment with pyrilamine, a histamine H1 receptor antagonist. These data suggest that thioperamide induces the release of neuronal histamine, which in turn stimulates both H1- and H2-receptors to produce the anxiogenic effect. The stimulation of histamine H1-receptors may mediate the anxiety, while H2-receptors may play a role in masking the anxiogenic effect. Thus, the present study suggests the involvement of endogenous neuronal brain histamine in anxiety. In the biochemical study, a previous report showed that thioperamide accelerated the release of neuronal histamine in the brains of mice [Sakai et al., Life Sciences, 48, 2397-2404(1991)]. This study also demonstrated that thioperamide did not affect the turnover rate of noradrenaline, dopamine, or serotonin in the brains of mice, which indicates that thioperamide is a good pharmacological tool for accelerating the release of neuronal histamine in the brain. PMID- 8231649 TI - Hepatic injury and lipid peroxidation during hemorrhagic shock and resuscitation. AB - Resuscitation from hemorrhagic shock causes hepatic injury that is similar to the hypoxic injury caused by reperfusion after ischemia. This study was designed to describe the relationship between severe hemorrhagic shock, hepatic injury, and lipid peroxidation. Fasted Sprague-Dawley rats underwent shock (mean arterial pressure 40 +/- 5 mm Hg) for two hours followed by reinfusion of shed blood. Serum ALT levels increased during shock and gradually continued to increase for 24 hours after resuscitation. Lipid peroxidation was assessed by quantification of exhaled ethane and by liver content of thiobarbituric acid reactive substances (TBARS). Expired ethane was increased both during shock and after resuscitation. Hepatic content of TBARS remained at baseline levels during shock, but increased after resuscitation. The results suggest that severe, non-fatal hemorrhagic shock and resuscitation produces a modest hepatic injury that is accompanied by lipid peroxidation in the liver. PMID- 8231650 TI - The kinin released from high molecular weight-kininogen is responsible for inflammatory exudation in rats: detection of kinin-free-kininogen in the exudate by immunoblot analysis. AB - Kinin release and its involvement in inflammatory exudation were assessed by immuno-blot analysis of the kinin-precursor protein, high molecular weight kininogen (HK). HK consists of heavy (H) chain, bradykinin and light (L) chain. After bradykinin was released by plasma kallikrein, HK remains two-chain-kinin free form;, i.e., H-chain and L-chain link each other through a disulfide bond. By Western blot analysis using antibody recognizing the light chain of HK, a band of 110-kD mass, which corresponds to intact HK, was detected in plasma after SDS PAGE under reducing conditions, while a 46-kD band, corresponding to the light chain of HK, but no 110-kD band, was found in the exudate of rats with carrageenin-induced pleurisy at 3 hr as well as at 16 h. This result indicates that in the exudate most all of the HK molecules had released kinin to form kinin free-HK, whereas the HK in the plasma remained intact. On the contrary, low molecular weight kininogen (LK) in the exudate was mostly in its intact form. These results indicate that plasma kallikrein could be activated in the exudate to release kinin from HK, as it reacts exclusively with HK and not with LK, and they are also mostly consistent with the features of the kinin release from the exudate and the plasma. That is, no kinin was detected in the exudate when the latter was incubated with plasma kallikrein, whereas salivary kallikrein did release kinin, indicating that kinin had already been released from HK, but not from LK in the exudate. Immunoblot analysis of HK in the pleural exudate also demonstrated no kinin involvement in phorbol myristate acetate- or zymosan induced pleurisy, since no light chain band, but an intact HK band, was found in the exudates from these pleurisies. PMID- 8231651 TI - Clonidine lowers alpha-MSH-like immunoreactivity in human plasma. AB - In a group of seven healthy subjects, the effects of acute intravenous administration of clonidine, a selective alpha 2 receptor stimulator, on plasma alpha-MSH-LI concentrations were measured. In comparison with saline, clonidine (0.075 mg) significantly reduced alpha-MSH-LI concentrations, with a maximum fall between 30 and 60 min., followed by a return to basal concentrations at 120 min.; no significant variations in plasma ACTH and cortisol were seen. The precise mechanism of this effect is unclear. Our study suggests that separate regulatory mechanisms exist for the secretion of POMC related peptides in the corticotroph and melanotroph cells of the human pituitary gland. PMID- 8231652 TI - Pro- and anti-inflammatory properties of human recombinant IL-1 beta during experimental arthritis in rats: 2. Period-dependent effect. AB - The systemic effects of human recombinant Interleukin-1 beta (HrIL-1 beta) on hindpaw edema were determined in arthritis induced by human native type II collagen (CII) with muramyl dipeptide (MDP) both injected on day 0. Daily treatment with HrIL-1 beta (0.2 microgram sc) pretreatment, from D-1 (the day before MDP and CII were injected) to D3 significantly delayed the secondary inflammation in the uninjected left hindpaw, whereas the same treatment from D6 to D10 at the end of the "primary" inflammation, enhanced the volume of the left hindpaw. Treatment from D13 to D17 did not affect the "secondary" edema in the left hindpaw. Thus, HrIL 1 beta administration produces pro- or anti-inflammatory effects on a developing polyarthritis depending on when treatment is started and is most effective as an anti-inflammatory molecule when started at the peak of the the inflammatory reaction, as previously described. In view of these early findings, we have compared the effect of adding HrIL-1 beta along with MDP in the sensitization procedure on the time-course of CII-induced arthritis. No adjuvant effect of HrIL-1 beta was observed. On the contrary, HrIL-1 beta significantly decreased the signs of inflammation in the injected hindpaw during the secondary inflammation. In addition, the immune response to type II collagen was less in the group receiving HrIL-1 beta, maybe because of nonspecific increase of antigen clearance. On the other hand, the MDP sensitization procedure enhanced the incidence of CII arthritis and significantly worsened the clinical parameters in both primary and secondary inflammations. PMID- 8231653 TI - Divinyl ethers and hydroxy fatty acids from three species of Laminaria (brown algae). AB - Three species of brown algae, Laminaria sinclairii, L. saccharina and L. setchellii, have been investigated for the presence of oxylipins. From one, L. sinclairii, three new divinyl ether fatty acids have been characterized as methyl ester derivatives (methyl 12-[1'(Z),3'(Z)-hexadienyloxy]-6(Z), 9(Z),11(E) dodecatrienoate, methyl 12-[1'(Z),3'(Z)-hexadienyloxy]-9(Z), 11(E) dodecadienoate, and methyl 14-[1'(Z),3'(Z)-hexadienyloxy]- 5(Z),8(Z),11(Z),13(E) tetradecatetraenoate) by a variety of spectroscopic methods. In addition, one new [13(S)-hydroxy-6(Z),9(Z),11(E),15(Z)-octadecatetraenoic acid] and four known monohydroxy polyunsaturated fatty acids have been isolated from all three species as their methyl ester derivatives. The occurrence of these compounds in brown algae strongly suggests that these organisms possess an active lipoxygenase(s) with omega 6 specificity. PMID- 8231654 TI - Age-dependent accumulation of phosphatidylcholine hydroperoxide in the brain and liver of the rat. AB - Age-related changes in phosphatidylcholine hydroperoxide (PCOOH) content as an index for oxidative membrane lipid damage were determined by high-performance liquid chromatography using chemiluminescence detection. Brain and liver PCOOH content increased significantly in male and female rats with age. The brain PCOOH content of male 18-month-old rats was 4.4 times that of 1-month-old rats, and that of female 18-month-old rats was 3.5 times that of 1-month-old females. The liver PCOOH content of the male 18-month-old rats was 9.3 times that of the 1 month-old; and of the female 18-month-old rats was 4.7 times that of the 1-month old. PCOOH levels in heart and lung did not show age dependency. In both brain and liver (but not in heart and lung), the phosphatidylcholine content significantly decreased upon aging. The results indicate that oxidative deterioration, such as phospholipid hydroperoxidation, is prevalent in the membrane lipids of brain and liver of the rat due to aging. PMID- 8231655 TI - Modulation of phosphatidylcholine biosynthesis by peroxisome proliferating fatty acid analogues. AB - The modulation of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) biosynthesis by sulfur-substituted fatty acid analogues has been investigated in rats. We have compared the effects of two non-beta-oxidizable fatty acid analogues, 3-thiadicarboxylic acid and tetradecylthioacetic acid, which induce proliferation of peroxisomes, with those of the analogue tetradecylthiopropionic acid, which is a weak peroxisome proliferator. Repeated administration of 3 thiadicarboxylic acid for seven days resulted in increased hepatic concentrations of both PC and PE, but the PC/PE ratio was decreased. PC synthesis was increased, as evidenced by increased incorporation of [3H]choline into PC and an increased activity of cytidinetriphosphate (CTP): phosphocholine cytidylyltransferase. This was accompanied by a reduction in the pool sizes of choline and phosphocholine. The S-adenosylmethione/S-adenosylhomocysteine ratio (AdoMet/AdoHcy) was marginally affected, indicating no increase in the rate of methylation of PE to PC. Administration of tetradecylthioacetic acid also resulted in increased hepatic phospholipid levels, increased AdoMet/AdoHcy ratios and in slightly elevated activity of CTP:phosphocholine cytidylyltransferase. The most striking effect observed after tetradecylthiopropionic acid treatment was the development of fatty liver. The activity of CTP:phosphocholine cytidylyltransferase and the incorporation of [3H]choline into PC was reduced compared to 3-thiadicarboxylic acid treatment. Although the rate of methylation of PE seemed to be increased at an elevated AdoMet/AdoHcy ratio, this resulted in only minor changes in the hepatic PC and PE levels, and the PC/PE ratio remained unchanged. Furthermore, the hepatic levels of choline and phosphocholine were reduced in these rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231656 TI - Distribution of ubiquinone and ubiquinol homologues in rat tissues and subcellular fractions. AB - The oxidized (UQox) and reduced (UQred) forms of ubiquinone (UQ) homologues in rat tissues and subcellular fractions were analyzed to elucidate their distribution and physiological role. UQ-9 and UQ-10 were detected in all tissues studied, and UQ-9 was the predominant homologue. The total amount of UQox-10 and UQred-10 was 20-50% that of UQox-9 and UQred-9. The levels of these homologues were highest in heart with lesser amounts occurring in kidney, liver and other organs. In liver and blood plasma, the UQred homologue amounted to 70-80% of the total UQ (UQox + UQred = t-UQ). UQred was less than 30% of t-UQ in other tissues and blood cells. t-UQ was much higher in leukocytes and platelets in blood than in erythrocytes. In erythrocytes, t-UQ was exclusively located in the cell membranes. UQox and UQred were also found in all subcellular fractions isolated from liver and kidney in about the same ratio as UQred/t-UQ was present in the whole organ. The levels of UQox and UQred per mg protein in subcellular fractions from liver were highest in mitochondria, with lesser amounts present in plasma membranes, lysosomes, Golgi complex, nuclei, microsomes and cytosol. In the mitochondria, the outer membranes were richer in t-UQ than the inner membranes. In the Golgi complex, the light and intermediate fractions were rich in t-UQ when compared to the heavy fraction. The possible physiological role of UQox and UQred in tissues and subcellular fractions is discussed. PMID- 8231657 TI - Effect of dietary alpha-linolenic acid and its ratio to linoleic acid on platelet and plasma fatty acids and thrombogenesis. AB - The effect of dietary alpha-linolenic acid (18:3n-3) and its ratio to linoleic acid (18:2n-6) on platelet and plasma phospholipid (PL) fatty acid patterns and prostanoid production were studied in normolipidemic men. The study consisted of two 42-d phases. Each was divided into a 6-d pre-experimental period, during which a mixed fat diet was fed, and two-18 d experimental periods, during which a mixture of sunflower and olive oil [low 18:3n-3 content, high 18:2/18:3 ratio (LO HI diet)], soybean oil (intermediate 18:3n-3 content, intermediate 18:2/18:3 ratio), canola oil (intermediate 18:3n-3 content, low 18:2/18:3 ratio) and a mixture of sunflower, olive and flax oil [high 18:3n-3 content, low 18:2/18:3 ratio (HI-LO diet)] provided 77% of the fat (26% of the energy) in the diet. The 18:3n-3 content and the 18:2/18:3 ratio of the experimental diets were: 0.8%, 27.4; 6.5%, 6.9; 6.6%, 3.0; and 13.4%, 2.7, respectively. There were appreciable differences in the fatty acid composition of platelet and plasma PLs. Nevertheless, 18:1n-9, 18:2n-6 and 18:3n-3 levels in PL reflected the fatty acid composition of the diets, although very little 18:3n-3 was incorporated into PL. Both the level of 18:3n-3 in the diet and the 18:2/18:3 ratio were important in influencing the levels of longer chain n-3 fatty acid, especially 20:5n-3, in platelet and plasma PL. Production of 6-keto-PGF1 alpha was significantly (P < 0.05) higher following the HI-LO diet than the LO-HI diet although dietary fat source had no effect on bleeding time or thromboxane B2 production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231658 TI - Dietary fish oil inhibits human breast carcinoma growth: a function of increased lipid peroxidation. AB - Female athymic nude mice were implanted subcutaneously with human breast carcinoma MDA-MB231. Seven to ten days later, the mice were divided into groups and fed a purified diet containing the following types of fat (% of diet): (i) 20% corn oil (CO); (ii) 15% CO:5% fish (menhaden) oil (FO); (iii) 10% CO:10% FO; (iv) 5% CO:15% FO; (v) 1% CO:19% FO; and (vi) 1% CO:19% FO plus antioxidants (alpha-tocopherol acetate, 2000 IU/kg diet and tertiary butyl-hydroquinone, 2% of total fat). The linoleic acid levels (% of diet) of the groups were 12.0, 9.1, 6.2, 3.3, 0.9 and 0.9%, respectively. After 6-8 wk, the carcinomas were assessed for tumor volume (cm3) and assayed for thiobarbituric acid reactive substances (TBARS). Human breast carcinoma growth was suppressed in mice consuming FO diets without antioxidants as compared to mice fed CO; the greater the amount of dietary FO fed, the greater the carcinoma growth suppression (P < 0.05). The addition of antioxidants to the FO diet significantly (P < 0.05) reversed the FO induced carcinoma growth suppression. Concentrations of TBARS in the human breast carcinomas were increased in all the FO (without antioxidants) fed mice, compared to mice fed CO; the level of increase in TBARS was directly related to the increase in the level of FO fed (P < 0.05). The addition of antioxidants to the FO diet significantly (P < 0.05) reduced the concentration of TBARS in the breast carcinomas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231659 TI - Higher levels of plasma cholesterol sulfate in patients with liver cirrhosis and hypercholesterolemia. AB - An analytical method for the determination of cholesterol sulfate (CS) in plasma using gas-liquid chromatography was developed. We measured plasma CS concentrations in patients with liver cirrhosis and hypercholesterolemia as examples of disorders that involve aberrations in cholesterol metabolism. Patients with liver cirrhosis had plasma CS concentrations that were significantly higher than those of control subjects (444.6 +/- 51.7 vs. 253.0 +/- 24.6 micrograms/dL, mean +/- SE). The levels of other lipids were lower in cirrhotics, although the differences were not significant. There was no correlation between the levels of CS and sulfated bile acids in cirrhotic patients. CS levels in plasma were also higher in subjects with hypercholesterolemia (413.7 +/- 44.5 micrograms/dL); however, the ratio of CS to total cholesterol (TC) clearly differed between cirrhotics and hypercholesterolemic subjects (1.44 +/- 0.11 x 10(-3) vs. 3.31 +/- 0.63 x 10(-3); P < 0.05). Both in subjects with hypercholesterolemia and in healthy controls, the CS/TC ratio was similar and CS accounted for roughly 0.14% of the TC concentration. PMID- 8231660 TI - Phospholipids from the free-living nematode Caenorhabditis elegans. AB - The phospholipid and the fatty chain compositions of diacyl, alkylacyl and alkenylacyl glycerophospholipids of the free-living nematode, Caenorhabditis elegans, were investigated. The phospholipids were comprised of 54.5% ethanolamine glycerophospholipid (EGP), 32.3% choline glycerophospholipid (CGP), 8.1% sphingomyelin and 5.1% others. The most abundant fatty acid in CGP was eicosapentaenoic acid (20:5n-3). The fatty acids in CGP were more unsaturated than those in EGP. Alkenylacyl and alkylacyl subclasses accounted for 1.0 and 2.6%, respectively, of CGP and 14.0 and 19.6%, respectively, of EGP. At least 80% of the alkenyl and alkyl groups were 18:0 chains and the remaining were odd numbered chains. The potential presence of platelet-activating factor (PAF) was examined by bioassay, but PAF-like activity was not detected in the extracts of this nematode. PMID- 8231661 TI - Effect of some potassium selective crown ethers on the permeability and structure of a phospholipid membrane. AB - The effect of some new crown ethers on the cation efflux and phase transition parameters of dipalmitoyl phosphatidylcholine liposomes was studied. The effects were correlated with the lipophilicity of the crown ethers. The results indicate that the presence of two crown ring structures in one crown either molecule is a prerequisite for the increase of ion permeability of liposomes. The effective crown ethers decrease the temperature, enthalpy and cooperativity of the gel-to liquid crystalline phase transition. The crown ethers increase membrane permeability for potassium and, to a lesser extent, for rubidium and sodium. The ratio of permeability increase for potassium/rubidium significantly correlates with the lipophilicity of the crown ethers. PMID- 8231662 TI - Synthesis of ethyl arachidonate-19,19,20,20-d4 and ethyl dihomo-gamma-linolenate 19,19,20,20-d4. AB - Ethyl 5,8,11,14-eicosatetraenoate-19,19,20,20-d4 and ethyl 8,11,14 eicosatrienoate-19,19,20,20-d4 were synthesized by Grignard coupling of the methanesulfonyl ester of 2,5-undecadiyn-1-ol-10,10,11,11-d4 with 5,8-nonadiynoic acid and 8-nonynoic acid, respectively. The coupled products upon Lindlar reduction, followed by the preparation of their ethyl esters, yielded deuteriated ethyl arachidonate and ethyl dihomo-gamma-linolenate, which were completely characterized by 13C and 1H nuclear magnetic resonance and mass spectral analysis. PMID- 8231663 TI - Continuous monitoring of lipid peroxidation by measuring conjugated diene formation in an aqueous liposome suspension. AB - A method is described for the direct and continuous monitoring of lipid peroxidation in an aqueous suspension of sonicated liposomes. By means of ultraviolet difference spectroscopy using tandem cuvettes, the formation of conjugated dienes during liposome peroxidation can be followed. Using this technique, the effect of the fatty acid composition of liposomes on lipid peroxidation can be studied. The results show that both the extent and the time scale of lipid peroxidation are influenced by the fatty acid composition of the phospholipid liposomes. This was confirmed also by other methods, such as measurement of the formation of lipid hydroperoxides and measurement of the decrease in polyunsaturated fatty acids. The advantage of the method described is the direct and continuous monitoring of phospholipid peroxidation in an aqueous environment, without subsampling and extraction of peroxidation products into organic solvents. Using this experimental approach based on difference spectra the contributions from changes in liposome, CuSO4 and H2O2 concentrations are canceled, thus improving sensitivity. The method can be employed for measuring the susceptibility to peroxidation of membrane phospholipids from fatty acid modified endothelial cells. PMID- 8231664 TI - Isolated tumor growth in a surgically formed skin pedicle in the rat: a new tumor model for NMR studies. AB - We have developed a new tumor model, the skin flap pedicle, which is ideally suited for study by such techniques as magnetic resonance imaging (MRI) and spectroscopy (MRS) and positron emission tomography (PET) to evaluate changes in metabolism/physiology in tumors before and after treatment. Heretofore, results from such studies were generally obtained from tumors growing in the thigh or flank region and these sites of growth often compromised the quality of the results, because of the contribution of normal tissues such as muscle and bone. In an attempt to circumvent such problems, we developed a surgical technique for creating a tubular skin pedicle and subsequently transplanting tumors in the lumen of such pedicles. We have successfully grown Dunning Prostate tumors (R 3327) as PEDICLE-TUMORS (PED-TUM). This paper describes the surgical procedure for formation of the skin pedicle, implantation of the tumor, data on the growth and histology and discusses the application of the PED-TUM to research problems. We compare the NMR characteristics of the tumor grown in the traditional SC thigh position with that growth in the pedicle and demonstrate the utility and advantages of the new model for studies of tumor physiological function and structure. PMID- 8231665 TI - Reduced lipid contamination in in vivo 1H MRSI using time-domain fitting and neural network classification. AB - It is a well-known problem that metabolite maps, reconstructed from in vivo 1H MRSI data sets, may suffer from contamination caused by the presence of strong lipid signals. In the present investigation, the lipid problem was addressed by applying specific signal processing and data-analysis techniques, combined with pattern recognition based on the concept of the artificial neural network. In order to arrive at images, cleaned from lipid artifacts, we have applied our previously introduced iterative and noniterative time-domain fitting procedures. Furthermore, reduction in computational time of the image reconstructions could be realized by using information provided by a neural network classification of the spectra, calculated from the MRSI data sets. PMID- 8231666 TI - Respiratory triggered imaging with an optical displacement sensor. AB - Motion of abdominal organs with respiration is a major problem in NMR spectroscopy and imaging thereof. Triggering each phase-encoding step with respiration or gating a number of phase-encoding steps is one approach to the problem. The design of a sensor for small animal experiments has not been as simple. An optical device, implemented with polymer optical fibres is described, along with associated hardware and electronics which can act as a trigger for small animal NMR experiments. A brief description of a similar device for human application is also given. 2DFT spin-echo and B0 susceptibility images, both triggered and untriggered, are presented to validate the technique. PMID- 8231667 TI - Magnetic resonance methods for measurement of disease progression in rheumatoid arthritis. AB - Magnetic resonance imaging methods are described for measurement of disease activity in knee in rheumatoid arthritis patients. Measurements of cartilage thickness, joint effusion volume, and pannus volume have been made. The latter measurement relies on synthetic fractional enhancement images of the response to gadopentetate dimeglumine. PMID- 8231668 TI - Identification of the distribution of sugars in grapes using chemical shift selective NMR microscopy. AB - Images are presented which show that NMR microscopy can be used to determine the spatial distribution of sugars in grapes. The production of appreciable sugar gradients during the final ripening process was detected in parenchymatous tissues around the seeds, but these gradients disappeared in fully ripened fruit. PMID- 8231669 TI - Integrated 3D display of brain surface anatomy and MR spectral data. AB - We describe a method of displaying the relationship between MR spectral data and an MRI-derived three-dimensional (3D) model of the same subject's brain surface. In this way, biochemical abnormalities in the MR spectra can be localized with respect to specific gyral convolutions (e.g., those associated with movement and sensation), which are best identified on 3D brain models. This was accomplished by retrospectively registering spectral data and MR images, acquired with different head coils. The highly resolved MR images were used to identify the brain surface in the poorly resolved spectral data and to produce a 3D rendition of brain surface metabolite distributions. This was then integrated with an MRI derived 3D model of brain gyral anatomy. The method was tested on P31 spectral data from a phantom and from a human volunteer. PMID- 8231670 TI - The use of power images to perform quantitative analysis on low SNR MR images. AB - Zero-mean noise introduced into quadrature detected MRI signals is generally rectified by the reconstruction algorithm to give a nonzero background intensity in the displayed image. In low signal-to-noise ratio (SNR) images, this background will inflate region of interest (ROI) signal measurements, leading to improper T2 and diffusion fits. A method is described here which separates signal from noise by computing power images from traditional magnitude data. Parameters measured from such power images show closer agreement with true values that do those derived from magnitude images. Because the correction algorithm is the same for all pixel intensities, it can be used with regions of interest with heterogeneous values. PMID- 8231671 TI - Magnetic resonance imaging of pericardial malignant mesothelioma. AB - Magnetic resonance imaging of pericardial malignant mesothelioma in two patients is reported. Magnetic resonance imaging clearly depicted the tumor location and expansion, and was useful in delineating the anatomic extent of pericardial malignant mesothelioma. PMID- 8231672 TI - Mesenteric varices: evaluation with color flow Doppler and magnetic resonance angiography. AB - Varix formation is a well recognized complication of portal hypertension. Rarely, varices may form in the distribution of the mesenteric veins. We present a case of mesenteric varices evaluated by color flow Doppler ultrasound and magnetic resonance angiography. PMID- 8231673 TI - Transdiaphragmatic leakage of ascites in cirrhotic patients: evaluation with ultrafast gradient echo MR imaging and intraperitoneal contrast enhancement. AB - The utility of intraperitoneal instillation of Gd-DTPA and ultrafast MR imaging in the diagnosis of transdiaphragmatic leakage of ascites was assessed in two cirrhotic patients with right-sided pleural effusion. Contrast-enhanced serial snapshot-FLASH reliably demonstrated the communication between the peritoneal and the pleural cavities in both patients. In one case, a right-sided diaphragmatic defect could be exactly localized. PMID- 8231674 TI - MR imaging of oculomotor nerve neurilemmoma. AB - The authors present a proved case of neurilemmoma arising from the oculomotor nerve in which MR imaging, including enhancement by gadopentetate dimeglumine, dynamic study and three-dimensional gradient echo images, provided much more precise definition of the tumor. PMID- 8231675 TI - Tumor-selective contrast enhancing agent, Mn(III)meso- [tri(4 sulfonatophenyl)phenyl]porphine (MnTPPS3) PMID- 8231676 TI - Gradient coil design: a review of methods. AB - Methods of coil design for MRI gradient coils and other purposes are discussed. The conflicting requirements for satisfactory gradient coil performance are described and analyzed. Design methods are classified into discrete wire techniques and current density techniques. The advantages and disadvantages of each group of methods are described, and optimization methods are summarized. These include the target field approach, and constrained inductance and power minimization. Design for low fringe fields is described, including active shielding. PMID- 8231677 TI - Dynamic imaging with T2* contrast using U-FLARE. AB - Dynamic changes in T2* in the rat brain induced by the injection of Gd-DTPA are monitored using the U-FLARE sequence. Sensitivity to T2* is easily introduced into this sequence and may be freely varied. The images obtained display an adequate spatial resolution and contrast, for a temporal resolution of 1 s. Intensity-time curves and parametric images of regional cerebral blood flow (rCBV) are presented for three different blood pCO2 values. The results presented clearly demonstrate that U-FLARE is a viable method for dynamically measuring changes in T2*, and thus has application in imaging of both perfusion and functional activation. PMID- 8231678 TI - Time of flight renal MR angiography: utility in patients with renal insufficiency. AB - We studied the renal arteries prospectively in 16 patients with renal insufficiency using a combination of two-dimensional and three-dimensional time of flight magnetic resonance angiography (MRA). Results were compared with conventional angiography. All renal arteries were identified by MRA. Accuracy for classifying renal arteries into patent, moderately (30-70%) stenotic, severely (> 70%) stenotic, or occluded was 91%. With regard to the presence or absence of severe occlusive disease (> 70% stenosis or occlusion) the sensitivity was 100%, with a specificity of 93%. PMID- 8231679 TI - Evaluation of suspected musculoskeletal neoplasms using 3D T2-weighted spectral presaturation with inversion recovery. AB - This study evaluated the use of the Spectral presaturation with inversion recovery (SPIR) technique with T2-weighting in 43 pathologically proven cases of suspected musculoskeletal neoplasm. Both primary and secondary malignant neoplasms as well as benign neoplasms were studied. The MR features exhibited by this technique are discussed. The images were evaluated by two experienced MRI specialists and graded as to utility into one of four categories as compared to conventional T1- and T2-weighted sequences. In the majority of cases this technique was found to be helpful or extremely helpful. The most useful features of this technique were the elimination of chemical shift artifact, the improved ability to evaluate superficial lesions or the extension of lesions into subcutaneous fat, and substantially improved visualization of both bone marrow and bone cortex interfaces. T2-weighted, fat suppressed imaging proved to be a useful new tool for evaluating musculoskeletal neoplasms. PMID- 8231680 TI - Detection of viable myocardium in segments with fixed defects on thallium-201 scintigraphy: usefulness of magnetic resonance imaging early after acute myocardial infarction. AB - To determine if magnetic resonance imaging (MRI) can be used to detect tissue viability in segments with persistent 201T1 defects early following acute myocardial infarction, 24 patients underwent MRI and adenosine 201T1 single photon emission computed tomography (SPECT) imaging at approximately 6 days. Infarction was demonstrated on MRI using a velocity-compensated, T2-weighted spin echo pulse sequence. Wall thickening was assessed using a gradient-echo pulse sequence obtained in the same anatomic position. Viable myocardium was defined by MRI as a segment with increased signal intensity and preserved wall thickening. A fixed defect on the 201T1 SPECT images was defined as the absence of any redistribution 4 hours after the 201T1 infusion. Of 11 patients with redistribution on the 201T1 images in the infarction region, 10 (91%) had preserved wall thickening by MRI. Of 13 patients with fixed defects on the 201T1 images in the infarction region, 6 (46%) had preserved wall thickening by MRI. Of 7 patients with absent thickening, all had one or more segments with absent perfusion on redistribution imaging. Wall thickening tended to occur in patients who received thrombolytic therapy or who underwent revascularization procedures prior to imaging. The results of the present study suggest that spin-echo MRI with motion compensation can be used to identify viable myocardium in patients with fixed defects on 201T1 SPECT following acute myocardial infarction. PMID- 8231681 TI - Evaluation of left ventricular inflow and volume by MR. AB - Considerable progress has been achieved during the past few years in the evaluation of intracardial blood flow by MR. Similar to ultrasound Doppler, it is possible now to registrate parameters of left ventricular diastolic filling as E/A ratio, velocity time integral of A- to E-wave (A(area)/E(area)), or isovolumetric relaxation time index. A total of 24 patients (7 women, 17 men, aged 24-69 yr) were examined, 5 of whom suffered from a hypertensive heart disease, 5 from a coronary heart disease, and 7 from mitral valve stenosis of various degree; 7 showed normal findings. E/A-ratio, A(area)/E(area), as well as isovolumetric relaxation time (IVRT) were evaluated by two different examiners on the same day by MR and ultrasound-Doppler, showing correlation factors of .89 for E/A ratio, .67 for A(area)/E(area), and .76 for IVRT in comparing both techniques. Additionally, to test the potential for making left ventricular function diagnoses from MR, time-volume curves for normal cases and patients with hypertrophic and dilated cardiomyopathy were elaborated by a computer-assisted 3D imaging technique. This technique showed good concordance with the results from cardiac catheterization or radionuclide ventriculography. PMID- 8231682 TI - MR imaging of fat-containing tissues: valuation of two quantitative imaging techniques in comparison with localized proton spectroscopy. AB - Since lipid protons, consisting mainly of triacylglycerols (TAG), are rather mobile, magnetic resonance imaging (MRI) is ideally suited for the examination of fat-containing tissues such as bone marrow. In contrast to water protons, however, lipid protons are chemically distinct and give rise to at least eight resonance peaks with different T1 and T2 relaxation times in the 1H spectrum. This is why the characterization of fat-containing tissues by quantitative MRI is much more difficult than that of most other tissues. In our study we wanted to examine the accuracy and the potential of a 1H chemical shift imaging (CSI) technique and a multiple spin-echo imaging (MSEI) technique. A stimulated-echo (STEAM) sequence for spatially localized proton spectroscopy was used as the reference method. In the first part of this paper, we describe quantitative imaging experiments which were performed to assess the accuracy of the fat-water separation according to the Dixon method and the bi-exponential decomposition of the MSEI data. For that purpose, we used a two-compartment phantom filled with either an aqueous Gd-DTPA solution and vegetable oil or with two different aqueous Gd-DTPA solutions, respectively. The analysis of the 1H CSI data revealed that the presence of non-methylen protons in neutral fats leads to a slight under estimation (of about 15%) of the relative fat fraction. The error is described theoretically and verified quantitatively by STEAM measurements. The bi exponential analysis of the transverse relaxation data, on the other hand, yields reliable T2 values if the relative proton density of both components is higher than 15%. IN the second part of our investigation, the same techniques were applied to acquire data from the subcutaneous fatty tissue, the femoral head, and the lumbar vertebrae of three healthy volunteers. In the bone marrow spectra, only two broad resonances could be resolved; they were superpositions of diverse molecular groups with different T1 and T2 relaxation times. In these cases, localized proton spectroscopy does not provide additional information with respect to 1H CSI. The MSEI data of the three examined fat containing tissue regions were adequately fitted by a bi-exponential function despite the fact that there were much more chemically distinct protons present in fatty tissues. PMID- 8231683 TI - Reliability of in vivo volume measures of hippocampus and other brain structures using MRI. AB - Volume reductions of the hippocampus are associated with Alzheimer's disease, schizophrenia, and epilepsy. We used clinically available MRI methods (2D acquisition; inversion recovery and calculated T2 images; 3 mm contiguous slices) that optimize image contrast, quality, and resolution and standardized positioning protocols to maximize the in vivo accuracy (test-retest reliability) of brain volume measurements in volunteers who were scanned two or three times. Volunteers were scanned in the same MRI instrument (intrascanner reliability) as well as in two different instruments (interscanner reliability). A single rater obtained brain volume measures of seven contiguous slices centered on the anterior commissure. The in vivo intrascanner reliability for measures of anterior hippocampus and ventricular volumes was very good, with reliability coefficients [intraclass r (rxx)] ranging between .855 and .997, and a median coefficient of variation (CV) of 6.4%. Reliability was good for amygdala (rxx of .740 and .764) and for total frontal and temporal lobe volumes and white matter volume measures (rxx ranging between .640 and .823, median coefficient of variation was 3.2%). Overall, interscanner reliability was also good. We discuss the implications of our results relative to the possible clinical utility of hippocampal quantification and the feasibility of prospective studies aimed at quantifying progressive neurodegeneration. PMID- 8231684 TI - Child abuse through the eyes of the teacher. AB - There are strict guidelines for dealing with suspected cases of child abuse in Great Britain. Teachers all know this but are often reluctant to call in the authorities, and see a stable family relationship broken, for what might be a trivial misdemeanour. With the help of several examples of pupils in a school in the north of England, and of experiences over several years of teaching, this article presents a variety of examples of abuse and shows how the author dealt with different experiences and problems. PMID- 8231685 TI - Evaluating the best interests of the child--a model of multidisciplinary teamwork. AB - This article describes the rationale, goals and procedure of a working model of multidisciplinary teamwork in conducting clinical evaluations of children and families and preparing expert testimony. Over the past five years, the team has evaluated over 60 cases involving child abuse and neglect, and questions of parental ability. Teamwork can reduce the distorting effects of personal biases, beliefs and countertransference issues. The contribution of integrating different theoretical viewpoints (theories of development, object relations, family systems, etc) is discussed. Recommendations include: (a) separation of roles of expert witness and therapist: (b) utilization of different diagnostic tools (psychodiagnostic tests, observations of interaction, joint clinical interviews, etc); (c) value of examining the feasibility of the recommendations with parents, care givers and community workers; and (d) strategies of dispute resolution and attainment of parental consent, utilizing the effects of potential judicial power and focusing on the best interests of the child. PMID- 8231686 TI - Involuntary outpatient commitment: ethics and problems. AB - The 1991 Israeli Mental Patients Law permits involuntary outpatient commitment (IOC) in an attempt to lessen the problem of recidivism of chronic patients who do not comply with medical treatment after discharge, relapse and are readmitted. The history of IOC in its different forms in several states in the United States is reviewed, with emphasis placed on indications and enforcement. Research assessment of the efficacy of IOC is discussed. The Israeli law is compared with the American law. Three case reports are presented. PMID- 8231687 TI - Ethical and moral dilemmas in the treatment of an abusive parent--the occupational therapy perspective. AB - Treating patients who physically abuse their children creates in the therapist moral and ethical dilemmas which challenge his or her ability to maintain a professional conduct and attitude. In such cases the therapist usually finds himself or herself confronted with the following dilemmas: (a) treatment of a patient who experiences therapy as a no-choice situation; (b) treatment of a patient whose conduct is in conflict with the moral values of the therapist; (c) the obligation of the therapist to report the abusive acts to the authorities versus his or her duty to protect the patient's right to confidentiality; and (d) the therapist's loyalty to the patient's welfare, when it is in conflict with the abused person's welfare. These issues are illustrated through the following case study examined in the context of occupational therapy (OT): A 26-year-old married woman, mother to a four-month-old infant was referred to therapy after causing severe burns to her daughter's hands. As a child the patient experienced physical abuse by her parents. The patient viewed treatment as her only possible means of keeping her child. On the other hand, she was suspicious of verbal therapy, as she assumed that the contents might incriminate her. Occupational therapy was therefore a major therapeutic modality used to assess and improve her disturbed psychosocial occupational performances. The article describes the way in which the OT approach helped overcome and resolve the moral and ethical dilemmas raised in the case. PMID- 8231688 TI - The selling of mental health services: some ethical reflections. AB - Since the introduction of public mental health services in Israel, the main principle of our work has been to provide equal and free of charge health services to all patients. We were proud of our ability to provide optimal treatment to all patients in all our facilities, regardless of cost or status of insurance. During the last decade, the cost of providing good quality public health services, including mental health services, has constantly increased, and the system has reached a state of financial distress resulting in insufficiency and inability to perform properly. In order to maintain the level of mental health services, the health authorities started planning a system of payment for various mental health services which, until now, were supplied free of charge. This change of policy and attitude towards the population in need poses severe ethical and practical questions and problems. It is questionable that the amount of income ensuing from the sale of mental health services and whether a relatively small financial profit justify possible injury of the population in need of these services, especially the sicker and weaker members of it. This article raises some ethical doubts involved in charging money for psychiatric services that are given to this special group of the mentally ill, and claims that the feasibility of selling services in this area of public health should be reinvestigated. PMID- 8231689 TI - Caring in the health care professions--a moral or a professional stand: perceptions of caring of female occupational therapists. AB - This article presents research that explores how female occupational therapists define and experience their professional caring role. The research analyses the relation between female occupational therapists' self-perception, their moral stand and their professional behaviour as patients' advocates. Caring for others, helping people in need, being responsible for the good of others; these and other calls of duty have been a major theme in defining the roles of occupational therapists as well as of other health care professionals. In the present study, seven female occupational therapists from the New York metropolitan area participated. For the collection of the data the researcher interviewed each of the participant, using in-depth ethnographic interviews. From the research main theme named 'self-perception as a caring person', four sub-themes were identified: (a) the perception of being a caring person; (b) being a caring person leads to becoming an occupational therapist; (c) being available to patients; and (d) protecting and helping patients. In this article the first three sub-themes are briefly described. The focus is on the fourth one. It analyses how the interviewees' self-perceptions as caring individuals are central to their moral decision-making process and to their involvement in patients' advocacy as part of their role and functions in occupational therapy. PMID- 8231690 TI - Releasing the treating psychiatrist from confidentiality. AB - Confidentiality is one of the oldest and most universal traditions of medicine. This rule is beneficial to the patient, who can disclose his or her secrets to his or her therapist without reluctance, to the physician, who enjoys a privileged relationships with his or her patient and to society at large, interested in public health and unrestricted access of all citizens to treatment. Nevertheless, each one of these beneficiaries claims the right to break this rule. The patient considers that the confidence is his or hers and may be disposed of at his or her discretion. The psychiatrist claims that he or she cannot remain silent and passive when the absence or delay of treatment of a reluctant patient can result in a disaster. Finally, society advocates the existence of a hierarchy which places the protection of society above the interests of the individual. Originally, breaches in confidentiality allowed by the legislator were chiefly intended to enable the patient to enjoy his or her rights or to facilitate treatment. Moreover the physician was encouraged to limit released data to those which were necessary for the purpose stated in the request for information. Unfortunately, the extent of the breaches is increasing. Some of the exceptions, like the obligation to report child abuse or imminent danger to others (Tarasoff) are usually readily accepted. This article details cases which give concern inasmuch as the disclosure of information exposes the patient to diverse restrictions. For example the right to confidence is waived by the patient under external pressure as a condition to obtain employment, access to military service or a driver's licence.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231691 TI - Breaking confidentiality survey. AB - Several Israeli laws require therapists to violate confidentiality. Psychiatrists, clinical psychologists and social workers were asked about their decisions to break confidentiality concerning information revealed in psychotherapy. No differences were found in confidentiality between private practice and public services. No differences were found between the three professional groups. Many therapists claim they do not report sensitive information revealed in psychotherapy, and prefer dealing with the information in therapy. Therapists were less inclined to break confidentiality about past (as opposed to present) sexual abuse. Therapists were less inclined to break confidentiality when sexual abuse was revealed in psychotherapy by the abuser than when revealed by the victim. PMID- 8231692 TI - Ethical issues in family law. AB - This article focuses on a few key ethical issues which arise in the context of family law. One of the issues faces mental health practitioners who are often engaged as expert witnesses in family law disputes. In another realm, family law attorneys face several fundamental ethical dilemmas in the course of their work. The client's right to appropriate treatment will be considered in the light of multidisciplinary services. On a broader level, the courts need to examine the ethical implications of the commonly used guideline in judicial process, 'the best interests of the child'. When these ethical issues are considered, it becomes clear what changes need to be made in individual practice, professional standards and social policy. The underlying premise is based on the knowledge we have gained in recent years through research and clinical experience with divorcing families. It is suggested that a radically different approach to society's handling of divorce is needed. Denmark's administrative handling of these problems is presented as a working alternative model. PMID- 8231693 TI - Informed consent in demented patients: a question of hours. AB - Dementia, and especially Alzheimer's disease is today the subject of endless research, most of it including some kind of prolonged physical treatment. When considering such treatment, one first has to deal with the quality and validity of the consent of the demented patient suspected of impaired judgment. The term 'informed consent' involves at least two types of decisions: one regarding short term intervention like a surgical procedure; the other concerns long-term intervention, like treatment by a new kind of drug intended to improve memory. In the latter case, the informed consent attributed to the patient raises both clinical and legal questions regarding validity of the continuing authorization given to the physician--a question which should also be examined in terms of time in fluctuating conditions, like dementia. What are the tests required to determine competency? What determines the grade of competency required in different types of research with different benefit-risk ratios? Do the changes in the cognitive function of the patient during the research demand constant assessment of competency. Where is the 'cut-off' point that dictates reassessment of the patient's consent and who is legally and practically responsible to identify this condition and decide whether to proceed? Finally, what are the alternative procedures that might simplify the decision-making process in fluctuating conditions without affecting the rights and autonomy of the patient? This article proposes several possible solutions to these legal and ethical dilemmas. PMID- 8231694 TI - Medical decisions under uncertainty. AB - The court applies the criteria of the reasonable doctor and common practice in order to consider the behaviour of a defendant physician. The meaning of our demand that the doctor expects that his or her acts or omissions will bring about certain implications is that, according to the present circumstances and subject to the limited knowledge of the common practice, the course of certain events or situations in the future may be assumed in spite of the fog of uncertainty which surrounds us. The miracles and wonders of creation are concealed from us, and we are not aware of the way and the nature of our bodily functioning. Therefore, there seems to be no way to avoid mistakes, because in several cases the correct diagnosis cannot be determined even with the most advanced application of all information available. Doctors find it difficult to admit that they grope in the dark. They wish to form clear and accurate diagnoses for their patients. The fact that their profession is faced with innumerable and unavoidable risks and mistakes is hard to swallow, and many of them claim that in their everyday work this does not happen. They should not content themselves by changing their style. A radical metamorphosis is needed. They should not be tempted to formulate their diagnoses in 'neutral' statements in order to be on the safe side. Uncertainty should be accepted and acknowledged by the profession and by the public at large as a human phenomenon, as an integral part of any human decision, and as a clear characteristic of any legal or medical diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231695 TI - The criminal liability of the mental patient in Jewish law (Halacha). AB - The legal status of the mentally impaired has been dealt with in Halachic literature since the third century. This article presents four Halachic viewpoints regarding the exemption of the mentally ill patient from legal responsibility in tort and criminal law: lack of mental awareness; lack of free will; lack of judgment; categorial personal status. Halachic legislations with regard to the liability of the insane predate the criteria as applied by modern psychiatry. The similarities between the Halachic and contemporary medicolegal approaches are presented. PMID- 8231696 TI - Who is to protect the rights of hospitalized mental patients? AB - The 1991 Treatment of Mentally Sick Persons Act contains a large section on the rights of hospitalized mental patients. However, mental patients are often ignorant of their rights, have difficulties gaining access to outside support and are hence unable to materialize their rights and protect them. Who is to protect their rights? The various individuals and groups which play a role in protecting the civil rights of mental patients are listed and discussed: patients themselves, family members, human rights organizations, legislators, courts, the state comptroller, the media and therapists. The introduction of another institution, non-existent in Israel, a 'patient advocate' (ombudsman) is suggested and discussed. PMID- 8231697 TI - Human rights: the right to receive treatment and care. AB - In the debates leading to the redesigning of the Mental Health Act in 1983 in the United Kingdom much pressure was exerted by human rights activists and other parliamentary pressure groups to ensure that patients could not be detained or treated 'against their will' without considerable--and at times undesirable- safeguards. This has led to difficulties over 'informed consent' and the setting up of the Mental Health Act Commission. The following examples are given: (a) A case dealing with injuries sustained by a patient while absent from the hospital while under a detention order. (b) The inability for psychotics to receive delayed release injections whilst in community care. (c) The loss of facilities for admission, and the resulting problems of homelessness arising from the hospital closure policy. PMID- 8231698 TI - Suspension pending inquiries: an English medical phenomenon? AB - The suspension of doctors in the National Health Service by health authorities, often for long periods for what appear to be trivial reasons, while inquiries into allegations are pending seems to have become an increasing problem in the United Kingdom. In some cases the hearing has been conducted years after the suspension. This situation caused the Society of Clinical Psychiatrists to establish a study group with the aim of producing a fact-finding report into what appeared to be a growing abuse of procedures. This article presents a summary of the study group's findings and the lessons to be learned from this disturbing bureaucratic phenomenon. PMID- 8231699 TI - The revision of the mental health commitment law in Belgium: the medical viewpoint. AB - Recently, the 1850 commitment law in Belgium has undergone dramatic revisions. These changes were implemented mainly at the request of law reformers who were concerned about the implications for civil liberties of institutionalization and who took into account the modern psychiatric treatment successes. Despite the fact that this legislation has promoted better progress in comparison with the original one, it has only been approved by the legal profession. The medical profession on the contrary is very critical and considers that this new measure gives few possibilities for the management of the doctor-patient relationship. The aim of the authors is to discuss, from a medical viewpoint, the advantages and disadvantages of the new law and to deliver some practical and ethical considerations. PMID- 8231700 TI - The expert opinion of the child and adolescent psychiatrist: a review of 140 lawsuits of minors involved in accidents. AB - One hundred and forty cases of claimed psychic trauma of a minor following an accident were reviewed. In two-thirds of the cases the injury followed a road accident. Other causes of injury were burns, animal bites, falls, etc. Injury 'by proxy' warranted compensation if psychic trauma was proven. Some of the more complex ethical, philosophical and psychiatric issues which confront the expert are reviewed such as credibility of the complaints, overexposure of a child to medical scrutiny, temporary versus permanent psychic damage, etc. PMID- 8231701 TI - The status of the mentally ill in Jewish law. AB - The Jewish law goes into great detail discussing the status of the mentally ill. There are many aspects to this question over and above the legal aspects of such a person's rights, obligations, doing business, etc. What is the Halachic approach to a mentally ill person in general? Is this person subject to the code of Jewish law the same as the normal Jew? Should we make an effort to help this type of person fulfil the commandments and prevent him or her from transgressing them or perhaps since such a person is incapable of controlling his or her behaviour, there is no purpose in these efforts? Marriage and divorce are other serious issues to which the Jewish law gives special attention in this context. Marriage must be entered into by a rational and judicious person or the act will not be valid. A very serious problem arises when a husband is mentally ill and due to that halachically cannot divorce his wife and she remains an Agunah. The situation is more complicated as the definition of mentally ill encompasses a broader spectrum of cases. Which psychiatric disorders come under the definition of a mentally ill person who is unable to control his or her behaviour? Which symptoms attest the inability of a person to enter into marriage or to grant a divorce? The Talmud discusses these matters in several places and the Halacha bases its rulings on their conclusions. PMID- 8231702 TI - Protection of hospitalized mental patients against sexual assault and abuse. AB - According to Israeli law the medical administration of a psychiatric hospital is responsible for the welfare, both physical and mental, of patients hospitalized in such a hospital and has therefore to take all measures apt to protect such patients against sexual assault and abuse. Any act of commission or omission which fails to comply with this duty may result in the administration, including its members personally, having to bear the responsibility, according to both penal and civil law, for the consequences suffered by the patient owing to a breach of their duty as mentioned above. Hospitalization of mental patients in absolutely separate wards for men and woman is the best means to ensure this protection. Therefore it is the duty of the medical administrator to provide hospitalization in separate wards for men and women. PMID- 8231703 TI - The multiple personality and the criminal law. AB - Multiple personality disorder (MPD) is a growing phenomenon in the United States. The number of cases have mounted following the celebrated cases of 'Three Faces of Eve' and 'Sybil' that have been popularized over the past several decades. How can--or should--the criminal law deal with multiple personality? Should the defendant suffering MPD be deemed competent to stand trial, or be able successfully to assert the insanity defence, or diminished capacity or automatism? Does MPD qualify as a mental disease or defect under the test of criminal responsibility, which would call for commitment upon a finding of not guilty by reason of insanity? What is the effect of MPD on cognition or control? These issues are discussed. PMID- 8231704 TI - The development of Israeli legislators' attitudes towards the rights of the mentally ill. AB - This article gives an overview of the Israeli legislators' handling of the rights of mentally ill persons. The developments and lacunae in the successive legislative provisions since 1955 until 1991 on the subject are critically examined. The role behind the scenes of politicians in shaping mental health legislation is highlighted. The article concludes with some proposals relating to future developments in the protection of the rights of the mentally ill. PMID- 8231705 TI - Patients and relatives--a human rights perspective on the legal regulation of intimate relationships. AB - This article examines the debate between advocates of patients' rights and relatives' rights through a study of the new Israeli Mentally Sick Persons Law of 1991. It indicates a duality in the legal treatment of relative-patient relations. On the one hand, relatives are assigned with the role of patients' de facto guardians. On the other hand, patients could be involuntarily hospitalized in order to protect their relatives' right to a normal way of life. Some attempts to resolve this apparent incoherence are discussed and the limitations imposed by the freedom principle are identified. It is argued that the focus should be upon the realities of an interpersonal relationship and not on some presupposed classification which has small legal relevance. PMID- 8231706 TI - Fatal asthma: where did we go wrong? AB - Vivid descriptions of life-threatening asthma attacks have persisted into modern medicine since antiquity. Asthma is characterized by airway hyperresponsiveness- a condition of an exaggerated bronchoconstriction in response to a variety of stimuli (irritants and triggers). Over the past decade it has become clear that in asthma significant inflammation occurs in the airways. Airway hyperresponsiveness and the clinical asthma symptomatology may be at least in part dependent on the inflammation generated by cell mediated immune mechanisms, including T cells, macrophages as well as mast cells and eosinophils in the airways. However, the recent advances in the understanding of the pathogenesis and pathophysiology of asthma, new modes for the diagnosis of the disease and the development of various therapeutic options have not been accompanied by increased survival rate. On the contrary, a gradually rising death rate from asthma has been reported worldwide. The reasons for this are not completely understood. Controversy exists whether an increase in the prevalence or the case fatality rate or both are the main cause of the reported rise in asthma death rate. Genetic, environmental and psychological factors have also been implicated. One has to consider the poorly controlled state in which some patients arrive at emergency wards, the possible delay in adequate therapy due to psychological and socioeconomic factors and also the information implicating iatrogenic factors among the possible explanations for increased asthma mortality. It is imperative to improve patient and family education to increase awareness. Conflicting results in the medical literature regarding the medical therapy of asthma impose a great responsibility on the practising physician. It is necessary to improve physician knowledge of the modern therapeutic strategies in asthma to gain better control of the disease. Pending further clarification as to optimal management of asthma, and forthcoming new and novel treatment, management should rely on current guidelines, emphasizing the role of antiinflammatory therapy in severe asthma. It is desirable to improve emergency services to deliver expert care in life-threatening conditions as soon as possible. PMID- 8231707 TI - General hospital psychiatry: implications for occupational therapy--one case study. AB - The hospitalization policy in general hospital psychiatric wards is for crisis intervention. The high cost in such a ward in Israel (in comparison with the cost in psychiatric hospitals) is justified since the patient is treated both physically and mentally. Some cases include patients who suffer from chronic diseases and aggravation of their physical condition causes severe mental reactions. In these cases, the short-term psychiatric admission is often in conflict with the condition of the patient and the desired treatment programme. The occupational therapist whose professional values are founded on the quality of life and his or her holistic approach to the patient, often has to deal with a serious dilemma: hospital policy on one hand and the patient's needs on the other. This dilemma is presented in its most acute form in the case study discussed in this article. PMID- 8231708 TI - The incident in Kure--a 40-year follow-up of post-traumatic stress disorder. AB - A traumatic incident involving Allied Occupation Forces in Japan in 1947 came to light in the course of pension applications by three Australian ex-servicemen. They had been present when a 'fire fight' took place between Australian and Indian soldiers based near Kure, apparently resulting in the death of two Sikhs. The emotional consequences had been suppressed, as had details of this incident, for 40 years. These cases provide a rare insight into the long-term consequences of traumatic events. The nature of chronic post-traumatic stress disorder and the necessity for evaluation of the contribution that traumatic experience can make to continuing psychopathology are discussed. PMID- 8231709 TI - Report of the committee assigned to investigate the protection of hospitalized mental patients against sexual assault and abuse--a critical review. AB - On 7 March 1991 a special committee was nominated by the Deputy Minister of Health to investigate the protection of mental patients hospitalized in Israel against sexual assault and abuse. The committee reached the conclusion that '[t]here were cases of sexual assault and abuse' but avoided examining the frequency of such cases. The committee interpreted the Israeli criminal law to mean that no patient has the ability to give consent to sexual relations, thus every incident of sexual relations with a patient has to be considered to be rape, that is the total responsibility rests on hospital staff. The inevitable recommendation was to convert all the psychiatric wards in Israel to unisexual wards. This article contains a critical review of the report. The main arguments brought forward are: (a) Most psychiatric wards in Israel are open, a fact that makes separation meaningless. Since the mixing of the wards both sexual and aggressive behaviour has been significantly reduced. (b) Many, if not most inpatients are voluntary and if brought to court would be found to be responsible for their actions, including their competence to give consent for sexual relations. (c) Patients, like everybody else, adjust their behaviour to the expectancies of society. The message that they are irresponsible will have a detrimental effect on their behaviour. (d) It is the staff's duty to prevent patients who may later regret their sexual behaviour from having sexual relations. This can be done without separating the wards.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231710 TI - Psychiatric hospitalization of dementia patients by commitment order in Israel. AB - Commitment order by a district psychiatrist is one of several modes of involuntary admission into a psychiatric hospital. Data regarding all the commitment orders by district psychiatrists in 1990 for elderly patients age 65 years or more were obtained from the national psychiatric case register. Demographic and clinical characteristics of patients diagnosed as ICD-9 senile organic psychotic conditions have been compared with the same characteristics of patients suffering from ICD-9 affective psychoses, schizophrenic disorders, paranoid states and transient organic psychotic conditions. The conclusions are: (a) Dementia patients are underrepresented in this sample of commitment orders; (b) from a demographic point of view dementia patients are no different from other psychiatric patients, age being the sole exception; and (c) from a clinical point of view dementia patients differ from other committed psychiatric patients in three ways: (i) their commitment is usually their first hospitalization; (ii) they are discharged from hospital within a two-month period; and (iii) they are referred for continuation of treatment in a non-psychiatric system. We conclude that psychiatric admissions under commitment order constitute a temporary solution for a very small group of dementia patients. PMID- 8231711 TI - The psychiatric patient as consumer. AB - The rocketing cost of good standard modern medicine and the constant strengthening of the consumers' voice are powerful factors behind the world-wide reorganization of medical services with the aim of improving cost-efficiency, quality and accessibility. The assumption of this article is that financial waste, redundancy and obsolence of medical systems are related to the non participation of patients in the evaluation of the services they consume. These problems are demonstrated and analysed using examples from psychiatry, where the patients and their families constitute a relatively silent consumer group. PMID- 8231712 TI - Aggressiveness in subjects referred for examination by court order: a retrospective study (1988-90) in the southern district of Israel. AB - Two hundred and eighty cases examined by court order by the district psychiatry office in the southern district of Israel in the past three years (1988-90) were critically reviewed. Analysis of the monthly distribution of criminal acts throughout the year shows a specific pattern of monthly occurrence of aggressive criminal acts (annual rhythm). This rhythm shows peaks in February and the summer months (June, July, August) and nadirs in March, October, November and December. The monthly occurrence of non-aggressive criminal acts showed a different pattern as compared with the annual rhythm of aggressive criminal acts. A correlation was found between the annual rhythm of aggressive criminal acts and the annual photoperiod (day length) rhythm. There was a significant positive correlation between these rhythms. We also analysed the age dependent distribution of aggressive criminal acts. A continuous age dependent increase in the percentage rate of aggressiveness was found. PMID- 8231713 TI - Listening to what the patient does not tell us. AB - In the intimacy of outpatient psychotherapy, the psychiatrist's ethical responsibility (burdensome at times) is always to place the emotional priorities of the patient ahead of the therapist's external and internal agendas. For example, the therapist's schedule (anticipated vacation), the therapist's interest in the patient for the sake of a publication, presentation, teaching, have to be subordinated to the emotional and somatic vulnerabilities and needs of the patient. The only sensor for this interaction is the professional conscience of the therapist. Excerpts from psychotherapy with six patients will highlight the dilemmas which each patient had which forced them to camouflage their hurts from themselves. An attempt will also be made to highlight ethical and professional dilemmas of the therapist. PMID- 8231714 TI - Psychiatric stigma and mental health professionals (stigmatizers and destigmatizers). AB - This article presents a definition of the term psychiatric stigma, its development, dynamics and role in society. The role that psychiatrists have played in the past and are playing in the present is described. The different factors that influence this role are elucidated culturally, professionally and economically. Examples are given of where psychiatrists act as destigmatizers and, on the other hand, as stigmatizers. The article stresses the point that psychiatrists should always be on the alert that in fulfilling their therapeutic role they can either stigmatize more or destigmatize their patients. Consideration should be taken in the overall approach so that the benefits to a patient are always more than his or her losses. Psychiatrists and mental health professionals must constantly be alert to ensure that they are acting in the best interest of the patient. PMID- 8231715 TI - South African psychiatrists' criteria for predicting dangerousness. AB - The prediction of dangerousness has emerged as one of the most controversial issues in forensic psychiatry. It is a value-laden and ambiguous concept which has not been adequately defined and operationalized by the law or psychiatry. The validity and reliability of psychiatric predictions of dangerousness have been brought seriously into question in the past few decades. The Booysen Commission appointed in South Africa relates to the problem. An exploratory survey to ascertain the variables which South African psychiatrists perceive as influencing their decisions about dangerousness and to compare these findings with those of previous research was undertaken. One hundred and thirty-eight psychiatrists were included in a survey by means of a questionnaire. Findings that clinicians' decisions were significantly influenced by patients' criminal/violent history were consistent with previous research. The article further evaluates the responses to the questionnaire variables and the degree as well as a study of detained 'dangerous' patients to assess accuracy of evaluations in practice. PMID- 8231716 TI - Patient-therapist sex: 'just say no' isn't enough. AB - This article focuses on the incidence of patient-therapist sex and the generally negative consequences for both parties; the characteristics of therapists against whom claims are laid; and the legal and ethical issues arising from such relationships. The authors conclude that prevention, in the form of more realistic and adaptive training programmes, is a far better approach to this problem than 'postvention' and the punishment of transgressing therapists. PMID- 8231717 TI - Is there sex after the prison door slams shut? AB - Despite the growing frequency of treatment-resistant STDs, North American literature and practice continue to stress repression and to evidence denial of prison sexuality. The authors outline their observations concerning prison sex, including officer-inmate sex. (Since this article was submitted, officer-inmate sex has received widespread public attention in Georgia, USA.) PMID- 8231718 TI - Mental illness and suicide in Israel. AB - The purpose of the present research was to assess the effect of mental illness on suicide completion risk. The sample consisted of 1,307 single and multiple suicide attempters whose first suicide attempt was reported to the Ministry of Health in Israel between the years 1972-1976. Each person was followed up retrospectively until 1983. The results indicate that about 18% of the multiple suicide attempters (repeaters) completed suicide. There was an equal distribution of the repeaters who were motivated to suicide by mental illness, depression and various social conflicts and difficulties. However, the frequency of suicide was greater among those who were motivated to suicide by mental illness and depression. A log linear model for prediction of suicide which included the motives to suicide with some other sociodemographic and clinical variables yielded a good fit. Mental illness was found to be one of the most important factors which affect suicide completion on both choice points, that is the first and second suicide attempt and especially from the second one. Based on these models we presented some calculations of suicide probabilities risk of the patients. Our findings show that almost two-thirds of the suicide attempts due to mental illness are in the high suicide risk. Theoretical interpretations of our findings, suggesting the impact of the labelling of mentally ill people and their possible unwillingness to enter a sick role, are suggested. PMID- 8231719 TI - More on Baby SAFE. PMID- 8231720 TI - Storytelling. A practitioner's tool. PMID- 8231721 TI - Comprehensive team care. PMID- 8231722 TI - When grandmothers take care of grandchildren. PMID- 8231723 TI - Latex use with infants and children: it can cause problems. PMID- 8231724 TI - Is breast-feeding possible after breast surgery? PMID- 8231725 TI - New genetic discoveries increase counseling opportunities. PMID- 8231726 TI - Tuberculosis: implications for patient teaching. PMID- 8231727 TI - TB: public health statutes. PMID- 8231728 TI - Time series analysis II: forecasting. PMID- 8231729 TI - Folic acid in the prevention of neural tube defects. PMID- 8231730 TI - Quantitative distribution of protein kinase C alpha, beta, gamma, and epsilon mRNAs in the hippocampus of control and nictitating membrane conditioned rabbits. AB - We used oligonucleotide in situ hybridization and film autoradiography to quantitate the distributions of protein kinase C (PKC) alpha, beta, gamma, and epsilon mRNAs in subregions of rabbit hippocampus. Levels of each of the hippocampal PKC isozyme mRNAs and patterns of their regional distributions were remarkably invariant between individuals. Within stratum pyramidale, the highest levels of PKC alpha mRNA were in the CA2 region, while PKC beta mRNA was maximally expressed in CA1, and PKC epsilon mRNA in CA3; PKC gamma mRNA was abundantly expressed throughout Ammon's horn. Previous experiments employing quantitative autoradiography for [3H]PDBU (Olds et al., Science, 245 (1989) 866 869) revealed an increase in membrane-bound PKC in the CA1 region of rabbit hippocampus up to 3 days following classical conditioning of the nictitating membrane response. We report here that there were no differences in levels of PKC alpha, beta, gamma, or epsilon mRNA between conditioned and control rabbits in any hippocampal region one day after training. These data are consistent with the hypothesis that PKC is post-translationally activated and translocated to the membrane during memory storage. PMID- 8231731 TI - Rapid increase of BDNF mRNA levels in cortical neurons following spreading depression: regulation by glutamatergic mechanisms independent of seizure activity. AB - Levels of mRNA for nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and the tyrosine kinase receptors trkB and trkC have been studied using in situ hybridization in the rat brain after topical application of KCl to the cortical surface (which induces spreading depression). Repeated episodes of spreading depression during 2 h caused a rapid and marked increase of BDNF mRNA levels in deep and, in particular, superficial cortical layers of the ipsilateral hemisphere (to 213 and 417% of control, respectively). Maximal levels were reached within 2 h after the cessation of spreading depression and at 24 h BDNF mRNA expression had returned to control values. Levels of BDNF mRNA were unaffected in the hippocampus, in areas outside the cerebral cortex and in the contralateral hemisphere. Furthermore, no change of the expression of mRNA for NGF, NT-3, trkC or the full length trkB receptor was detected at any time point. However, at 2 h after spreading depression there was an increased level (150% of control) in superficial cortical layers of mRNA hybridizing to an oligonucleotide probe detecting both truncated receptors lacking the tyrosine kinase domain and full length trkB receptors. Also one single episode of spreading depression gave rise to a significant increase of cortical BDNF mRNA levels (to 207% of control), which was attenuated (by 61%) after administration of the competitive NMDA receptor antagonist CGS 19755. The results provide evidence that mild brain insults associated with glutamate release and elevated intracellular calcium, such as spreading depression, also in the absence of seizure activity can lead to activation of the BDNF gene in cortical neurons. PMID- 8231732 TI - Structure of the human gene for the neural phosphoprotein B-50 (GAP-43). AB - The genomic DNA encoding the exons for the human neural phosphoprotein B-50 (GAP 43) was isolated using rat-based cDNA probes and oligonucleotides. Exons 2 and 3 were isolated from a genomic library, exon 1 was amplified by PCR on total genomic DNA. The gene consists of 3 exons and 2 large introns. The first exon encodes the N-terminal 10 amino acids of B-50 involved in membrane association of the protein. Exon 2 encodes the main part of the protein with the sites for protein kinase C-mediated phosphorylation and calmodulin binding, and includes a 10 amino acid residue insert not found in rodents. Exon 3 encodes the last 29 amino acid residues. The reported sequence extends the known cDNA structure to both the 5' and 3' ends. The 358 bp region upstream of the translational initiation codon, containing the main transcription starts, is purine-rich and does not include TATA or GC boxes. At the 3' end potential polyadenylation signals were found 510 bp and 584 bp downstream of the stopcodon in exon 3. The 5' end of the mRNA is heterogeneous in length, with primer extension products corresponding to a 5' untranslated region of 159 and 343 bases. Northern hybridizations, however, indicate that the majority of B-50 mRNA has a shorter 5' untranslated region, as was reported for the rat (Schrama et al., Soc. Neurosci. Abstr., 18 (1992) 333.4). The structural organization of the human gene is similar to that described for the rat (Grabczyk et al., Eur. J. Neurosci. 2 (1990) 822-827), and both translated and untranslated regions show a high degree of sequence homology to the rat gene. PMID- 8231733 TI - Expression of the mas proto-oncogene in the rat hippocampal formation is regulated by neuronal activity. AB - The mas proto-oncogene encodes a seven membrane-spanning G-protein-coupled receptor which is activated by angiotensins. In the postnatal and adult rat, mas mRNA is specifically expressed at high levels in hippocampal neurons. We report here using in situ hybridization and RNase protection that brief seizure episodes lead to a significant and transient increase in mas mRNA in the hippocampus. Increased levels of mas transcripts were detected 2, 4, and 6 h following seizure. By 24 h post seizure, baseline levels were detected. The presumed subsequent increase of the mas receptor protein may contribute to anatomical and physiological plasticity that is associated with intense activation of hippocampal pathways. PMID- 8231734 TI - Regulation of dopamine D2 receptor mRNA expression in the olfactory tubercle by cocaine. AB - The influence of acute and chronic cocaine treatment on the expression of dopamine D2 receptor (D2R) mRNA was examined in the rat basal ganglia by in situ hybridization histochemistry. No significant alterations in D2R mRNA levels were observed in the striatum or the nucleus accumbens following acute or chronic cocaine. In the olfactory tubercle, acute cocaine (20 and 40 mg/kg, i.p.) was found to decrease D2R mRNA expression. Chronic cocaine (20 mg/kg, i.p. x 15 days), however, produced an increase in D2R mRNA levels in this region which was detected 24 h, but not 7 days after withdrawal. Acute and chronic cocaine have different effects on the regulation of D2R gene expression in the olfactory tubercle. PMID- 8231735 TI - Time-course of changes in striatal levels of DA uptake sites, DA D2 receptor and preproenkephalin mRNAs after nigrostriatal dopaminergic denervation in the rat. AB - Changes in striatal dopamine uptake sites, D2 receptor and preproenkephalin (PPE) mRNA levels provoked by unilateral 6-hydroxydopamine-induced lesion of the nigrostriatal dopaminergic (DA) pathway were studied by quantitative autoradiography and in situ hybridization (ISH) in rats sacrificed at different post-lesion delays. The disappearance of DA terminals as visualized with the labelling of dopamine uptake sites with [3H]GBR 12935 became significant 36 h after the lesion and was almost complete at a delay of 7 days. PPE mRNA amounts significantly increase (+24%) already at the shortest delay studied (9 h after the lesion) while the labelling of the uptake sites on DA terminals was not affected. The time course increase of PPE mRNA levels was progressive until 21 days post-lesion where it reached its maximum (+132%) and remained stable up to the latest delay studied (60 days). Conversely D2 mRNA contents remained unchanged up to 5 days post-surgery and then increased relatively quickly since at 7 days post-lesion their levels were near (+21%) the maximum observed which was reached at 21 days post-lesion (+32%). This study suggests a time-dependent differential sensitivity to the degree of DA denervation of both major components implicated in the striatopallidal output. PMID- 8231736 TI - Loss of 5-HT1A receptor mRNA in the dentate gyrus of the long-term adrenalectomized rats and rapid reversal by dexamethasone. AB - Two months after bilateral adrenalectomy, 5-HT1A receptor mRNA labelling was decreased in the granular cell layer of the dentate gyrus but not in the pyramidal cell layer of Ammon's horn. Two month adrenalectomized rats given dexamethasone (10 micrograms/ml saline) 24 or 72 h before perfusion showed a progressive recovery in 5-HT1A mRNA labelling in the dentate gyrus. 5-HT1A expression may underlie hippocampal neuronal plasticity after long-term adrenalectomy. PMID- 8231737 TI - Localization of the cellular expression pattern of cdc25NEF and ras in the juvenile rat brain. AB - In this report, we demonstrate that the brain-specific ras nucleotide-exchange factor, cdc25NEF-B, is expressed in specific neuronal populations in the juvenile rat brain. Because cdc25NEF-B likely regulates one or more of the vertebrate ras proteins, H-, K- and N-ras, we also examined their levels of expression and pattern of expression in the juvenile rat brain. We find cdc25NEF-B to be highly expressed in the hippocampus, some deep nuclei, neocortex, and the granule cell layer of the anterior lobules of the cerebellum. Our observations suggest a functional link between cdc25NEF-B and H-ras in a neuronal signal transduction pathway. PMID- 8231738 TI - Sympathetic and parasympathetic ganglia express non-NMDA type glutamate receptors: distinct receptor subunit composition in the principle and SIF cells. AB - The presence of non-NMDA glutamate receptors in the rat sympathetic and parasympathetic ganglia was examined by immunocytochemistry using specific antibodies against AMPA-type excitatory amino acid receptor subunits (GluR1-4). Three kinds of antibodies specific to the GluR1, GluR2 and 3, and GluR4 subunits were used. The superior cervical ganglion and pterygopalatine ganglion were examined as representatives of sympathetic and parasympathetic ganglia. In the superior cervical ganglion, GluR1- and GluR2/3-like immunoreactivity was observed in most principal neurons and SIF cells. In contrast, GluR4-like immunoreactivity was not observed in the principal cells; however, SIF cells exhibited intense immunoreactivity of GluR4. In the pterygopalatine ganglion, the profile of the immunoreactivity was similar to that seen in the superior cervical ganglia. The subunit compositions between the principal cells and SIF cells were different, whereas the compositions in cell species involved in the autonomic ganglia, sympathetic and parasympathetic ganglia were identical. This suggests that glutamate is another important preganglionic transmitter together with acetylcholine, and the responses elicited in the principal cells and SIF cells might be different because of the difference in subunit composition. PMID- 8231739 TI - Elevated basal AP-1 DNA binding activity in developing rat brain. AB - Nuclear extracts from hippocampi, striata and hypothalami of postnatal day (P) 7 rats contained elevated basal levels of AP-1 DNA binding activity and c-jun protein, which decreased to the low basal levels observed in the adult by P21. In contrast to the AP-1 DNA binding complex in the adult brain, the fos-related antigens were not a major component of the P7 AP-1 DNA binding activity. PMID- 8231740 TI - On walking in America. PMID- 8231741 TI - Howard Raiffa and the discovery of decision analysis. PMID- 8231742 TI - Resistance to clinical computing systems. PMID- 8231743 TI - Using information technology to fight AIDS. PMID- 8231744 TI - Subjective probability. PMID- 8231745 TI - Mathemedicine in the Land of Odz. PMID- 8231746 TI - Visual programming languages. AB - Visual programming languages enable physicians and other computer users with little knowledge of programming to develop computer software. The physician uses a visual paradigm to "draw" the computer interface and then attaches short segments of computer code to buttons, menus, and list boxes. The programs can be written for DOS or Windows and can be used with various printers and input devices (a keyboard, mouse, or pen). This article describes the development of an example of a medical program, which calculates the treadmill score. PMID- 8231747 TI - Decision analysis: introductory lectures on choices under uncertainty. 1968. PMID- 8231748 TI - Using your computer's communications ability for education and fun. PMID- 8231749 TI - MacHeart: a model-based authoring system for CAI in cardiology. PMID- 8231750 TI - American College of Sports Medicine. Position Stand. Physical activity, physical fitness, and hypertension. AB - Hypertension is present in epidemic proportions in adults of industrialized societies and is associated with a markedly increased risk of developing numerous cardiovascular pathologies. There is a continuing debate as to the efficacy of aggressive pharmacological therapy in individuals with mild to moderate elevations in blood pressure. This has led to a search for nonpharmacological therapies, such as exercise training, for these individuals. The available evidence indicates that endurance exercise training by individuals at high risk for developing hypertension will reduce the rise in blood pressure that occurs with time. Thus, it is the position of the American College of Sports Medicine that endurance exercise training is recommended as a nonpharmacological strategy to reduce the incidence of hypertension in susceptible individuals. A large number of studies indicate that endurance exercise training will elicit a 10 mm Hg average reduction in both systolic and diastolic blood pressures in individuals with mild essential hypertension (blood pressures 140-180/90-105 mm Hg). Endurance exercise training also has the capacity to improve other risk factors for cardiovascular disease in hypertensive individuals. Endurance exercise training appears to elicit even greater reductions in blood pressure in patients with secondary hypertension due to renal dysfunction. The mode (large muscle activities), frequency (3-5 d.wk-1), duration (20-60 min), and intensity (50-85% of maximal oxygen uptake) of the exercise recommended to achieve this effect are generally the same as those prescribed for developing and maintaining cardiovascular fitness in healthy adults. Exercise training at somewhat lower intensities (40-70% VO2max) appears to lower blood pressure as much, or more, than exercise at higher intensities, which may be important in specific hypertensive populations. Physically active and fit individuals with hypertension have markedly lower rates of mortality than sedentary, unfit hypertensive individuals. Thus, it seems reasonable to recommend exercise as the initial treatment strategy for individuals with mild to moderate essential hypertension. A follow-up period should assess the efficacy of the patient's exercise program, and adjunct therapies should be implemented according to the individual patient's blood pressure and CAD risk factor goals. Individuals with more marked elevations in blood pressure (> 180/105 mm Hg) should add endurance exercise training to their treatment regimen only after initiating pharmacologic therapy. Resistive, or strength, exercise training is not recommended to lower blood pressure in individuals with hypertension when done as their only form of exercise training.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8231751 TI - Plasma lactate accumulation and distance running performance. 1979. AB - Laboratory and field assessments were made on eighteen male distance runners. Performance data were obtained for distances of 3.2, 9.7, 15, 19.3 km (n = 18) and the marathon (n = 13). Muscle fiber composition expressed as percent of slow twitch fibers (%ST), maximal oxygen consumption (VO2max), running economy (VO2 for a treadmill velocity of 268 m/min), and the VO2 and treadmill velocity corresponding to the onset of plasma lactate accumulation (OPLA) were determined for each subject. %ST (R > or equal to .47), VO2max (r > or equal to .83), running economy (r > or equal to .49), VO2 in ml/kg min corresponding to the OPLA (r > or equal to .91) and the treadmill velocity corresponding to OPLA (r > or equal to .91) were significantly (p < .05) related to performance at all distances. Multiple regression analysis showed that the treadmill velocity corresponding to the OPLA was most closely related to performance and the addition of other factors did not significantly raise the multiple R values suggesting that these other variables may interact with the purpose of keeping plasma lactates low during distance races. The slowest and fastest marathoners ran their marathons 7 and 3 m/min faster than their treadmill velocities corresponding to their OPLA which indicates that this relationship is independent of the competitive level of the runner. Runners appear to set a race pace which allows the utilization of the largest possible VO2 which just avoids the exponential rise in plasma lactate. PMID- 8231752 TI - The effect of prone immersion on bronchial responsiveness in children with asthma. AB - We studied the effects of prone immersion on pulmonary function in children with asthma. Twelve children with asthma were subjected to prone immersion (WET) and standing upright on land (DRY) under controlled conditions regarding temperature and relative humidity of the inhaled air, minute ventilation, respiratory frequency, tidal volume, and the concentration of carbon dioxide in the exhaled air. Eight minutes of isocapnic hyperpnea was performed in random order either in the swimming pool (prone) or on land (upright), with the temperature and relative humidity of the inhaled air kept at 20 +/- 2 degrees C and 10 +/- 2%, respectively. The average accumulated ventilation for the 8 min of hyperpnea was 3151 for the WET treatment and 3101 for the DRY (P > 0.1). Average heart rates were 101 +/- 11 and 115 +/- 17 beats.min-1 for the WET and DRY treatments, respectively (P < 0.05). The decrease in FEV1 (delta FEV1) was 26.3 +/- 16.7% after the WET treatment (compared with pretreatment FEV1), and 26.4 +/- 11.3% after the DRY treatment (P > 0.1 between WET and DRY delta FEV1), with similar trend (insignificant differences between treatments), for delta FVC, delta PEF 50%, and delta PEF 25-75%. It is concluded that airway hyperactivity is not alleviated by whole body prone immersion. PMID- 8231753 TI - Bone mineral density in elite junior Olympic weightlifters. AB - The purpose of this study was to examine the relationship of bone mineral density (BMD) to muscular strength in highly trained young male athletes in order to gain insights concerning the influence of heavy resistance training on BMD. Twenty five elite junior weightlifters (age, 17.4 +/- 1.4 yr) and 11 age-matched controls (16.9 +/- 1.1 yr) volunteered for this investigation. Measurements of BMD (g.cm-2) utilizing dual energy x-ray absorptiometry were obtained for the lumbar spine (L2-4) and the proximal femur (neck; trochanter, Ward's triangle). The BMD values for the junior lifters were found to be significantly greater at all sites for the junior weightlifters compared with their age-matched control group. The BMD values of the spine and femoral neck of the junior weightlifters when compared with adult reference data (i.e., 20-39 yr old men) were found to be significantly greater. Both simple and multiple regression analyses demonstrated significant relationships of BMD with strength accounting for 30-65% of the variance. These data suggest that in elite junior weightlifters, muscle strength, highly specific to the sport of weightlifting, has a major influence on BMD due to the influence of the chronic overloads experienced in training. PMID- 8231754 TI - Changes in diabetic urinary transferrin excretion after moderate exercise. AB - Microalbuminuria following submaximal exercise testing has been proposed for detecting renal abnormalities in diabetic patients. We compared urinary transferrin and albumin excretion between eight adults with insulin dependent diabetes mellitus and eight nondiabetic controls without microalbuminuria before and after a standardized exercise challenge of only moderate intensity for 20 min. Both groups were similar for age, sex, and METs expended during treadmill walking. Urinary excretion ratios of transferrin (UTER) and albumin (UAER) did not significantly increase for nondiabetic subjects. After exercise, UTER increased on average 207% in diabetic subjects (P = 0.009) and UAER increased 209% (P = 0.046). The percent increase in UTER appears to be a function of workload intensity, while the percent increase in UAER appears less dependent on the duration of exercise. A standardized treadmill challenge of moderate intensity easily differentiated changes in urinary transferrin excretion ratios between diabetic and nondiabetic subjects. Measuring transferrin excretion may be a more sensitive parameter than albumin in studies using urinary protein excretion as a response to a provocative exercise challenge. PMID- 8231755 TI - Oxygen uptake and heart rate relationship in persons with spinal cord injury. AB - The percent (%) peak oxygen uptake (VO2) and % peak heart rate (HR) relationships were determined in 13 persons with high (T1-T6) and 14 persons with low lesion (T7-T12) spinal cord injured paraplegia (SCI PARA) and 15 nonimpaired subjects during graded arm crank (AC) tests to exhaustion. Subjects were instructed to maintain a target cadence of 60 rpm on a modified electronically braked leg cycle ergometer. After 3 min of unloaded cranking, power output (PO) was increased by 8 16 W.min-1. VO2 and HR were determined via open-circuit spirometry and 12-lead ECG, respectively. Absolute HR and VO2 values for each PO were converted to % peak HR and % peak VO2 values. Linear regression slopes describing individual % peak HR and % peak VO2 relationships were calculated and compared between groups with one-way ANOVA. No significant differences (P > 0.05) were noted between the mean (+/- SD) regression slopes for persons with high lesion SCI PARA (1.48 +/- 0.21), persons with low lesion SCI PARA (1.48 +/- 0.26), and nonimpaired subjects (1.53 +/- 0.29). Regression equations derived using all data points within each group were as follows: High lesion SCI PARA: y = 1.3x-37.0, R = 0.85 Low lesion SCI PARA: y = 1.23x-30.9, R = 0.88 Nonimpaired subjects: y = 1.41x-46.2, R = 0.95 (y = % peak VO2, x = % peak HR). These equations are similar to those previously reported for nonimpaired men and women and cardiac patients during AC and leg cycle ergometry.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231756 TI - Inspiratory muscle performance relative to the ventilatory threshold in healthy subjects. AB - Inspiratory muscle performance, ventilation, and gas exchange were studied during exercise in healthy subjects to look for typical changes of pattern of contraction at the ventilatory threshold (VT). The steepening of the slope of carbon dioxide output (VCO2) vs oxygen uptake (VO2) at the VT was accompanied by a nonlinear increase of the mean rate of esophageal pressure development (Pes/TI) vs the esophageal pressure time index (PTIes) reflecting both the relative force (Pbreath/Pesmax) and duration (TI/TTOT) required for inspiration. The esophageal pressure time integral within one breath (Pbreath.dTI) was one of the best single predictors of the ventilatory equivalent for oxygen (VE/VO2) at the VT. Moreover, we presented inspiratory muscle load indices as a mirror image of breathing pattern, with the obvious advantage that the ventilation component can be compared with better established methods of presenting ventilatory output. Inspiratory muscle performance during exercise should link the increased metabolic rate to ventilatory output. We conclude that 1) there exists an inspiratory muscle threshold that is well correlated to commonly used gas exchange thresholds, and 2) the efficiency of ventilation and gas exchange during exercise could be linked to pressure and timing of inspiratory muscle contraction. PMID- 8231757 TI - Effects of high- vs moderate-intensity exercise on natural killer cell activity. AB - The effect of 45 min of high- (80% VO2max) vs moderate- (50% VO2max) intensity treadmill exercise on natural killer cell cytotoxic activity (NKCA) was investigated in 10 well-conditioned (66.0 +/- 1.9 ml.kg-1.min-1), young males (22.1 +/- 1.3 yr). Blood samples were taken before and immediately after exercise, with three more samples taken during 3.5 h of recovery, and analyzed for proportion of NK cells (CD3-CD16+CD56+) and NKCA. Exercise at 80% vs 50% VO2max resulted in a greater immediate postexercise increase in proportion of NK cells, followed by a 1-h and 2-h decrease below preexercise levels for both intensity conditions. NKCA rose significantly above preexercise levels following high- but not moderate-intensity exercise. For both exercise intensity conditions, NKCA tended to drop below preexercise levels by 1 h postexercise, rising back to preexercise levels by 3.5 h postexercise. When NKCA was expressed on a per-NK cell basis, however, no change relative to preexercise levels occurred following moderate-intensity exercise, while a significant increase occurred after 2-h recovery from high-intensity exercise. These data demonstrate that both high- and moderate-intensity exercise are associated with significant shifts in circulating proportions of NK cells which significantly influence interpretation of NKCA data based on assays using separated mononuclear cells. PMID- 8231758 TI - High intensity training-induced changes in skeletal muscle antioxidant enzyme activity. AB - These experiments tested the hypothesis that high intensity (interval) training is superior to moderate intensity (continuous) exercise training in the upregulation of antioxidant enzyme activity in skeletal muscle. To test this postulate, we examined changes in oxidative and antioxidant enzyme activities in rat skeletal muscle following 12 wk of either interval (6 x approximately 5-min intervals at approximately 80-95% VO2max) or continuous (45 min at approximately 70% VO2max) exercise training. Both continuous and interval training resulted in significantly elevated (P < 0.05) succinate dehydrogenase (SDH) and 3-hydroxyacyl CoA-dehydrogenase (HADH) activities in the gastrocnemius (G) and soleus (S) muscles compared with controls. SDH and HADH activities in the G and S muscles did not differ between the two exercise groups. Glutathione peroxidase (GPX) activity exceeded controls (P < 0.05) in only the interval trained S muscle. Soleus superoxide dismutase (SOD) activity was higher (P < 0.05) in both exercise groups compared with controls. No differences in SOD activity existed between interval and continuous trained animals. We conclude that when matched for oxygen cost, interval and continuous exercise training result in similar increases in SOD activity. However, high intensity interval exercise is superior to moderate intensity continuous exercise in the promotion of GPX activity in the S. PMID- 8231759 TI - Anthropometric growth patterns of high school wrestlers. AB - The purpose of this investigation was to examine the anthropometric growth patterns of high school wrestlers; 477 high school wrestlers volunteered as subjects for this study. The total sample was divided into four independent age groups: Age group 1 (AG1) = 14.00-14.99 yr (N = 38); AG2 = 15.00-15.99 yr (N = 130); AG3 = 16.00-16.99 yr (N = 163); and AG4 = 17.00-17.99 yr (N = 146). Thirteen anthropometric dimensions (seven diameters and six circumferences) were taken on each subject. To examine normal growth patterns, the anthropometric data were compared with values from a national representative sample of adolescent males. The results indicated that there were few differences between the wrestlers and the national sample for yearly changes in the anthropometric dimensions. These findings suggest that participation in high school wrestling, which typically includes repeated bouts of weight cycling, does not adversely affect normal anthropometric growth patterns. PMID- 8231760 TI - The physical condition of elderly women differing in habitual physical activity. AB - In a cross-sectional study, 50 elderly women (age 71.5 +/- 4.2 yr, mean +/- SD) participated in a battery of tests assessing several aspects of physical fitness. The women were selected based on tertiles of habitual physical activity as determined by a validated questionnaire 10 months ago. The tests comprised the following measurements: peak expiratory flow, flexibility of shoulder joint, flexibility of hip and spine, balance, reaction time, grip strength, manual dexterity, and endurance. Additionally, data were collected on height, body weight, and systolic and diastolic blood pressure. A questionnaire was used to evaluate subjective fitness and general subjective health. Results indicated that physically more active elderly women tend to have better results on most tests. Body weight and body mass index, flexibility of the hip and spine (assessed using a sit-and-reach test), and endurance on a walk test were significantly better in the more active women. Test results are confirmed by subjective evaluation by the participants. PMID- 8231761 TI - Preferred and energetically optimal gait transition speeds in human locomotion. AB - A widespread assumption of previous researchers is that the gait transition during human locomotion takes place at speeds that minimize metabolic energy consumption. The primary purpose of this investigation was to determine, by direct measurements, whether changing gaits is actually an energy saving mechanism. The secondary purpose of the experiment was determine whether the sense of effort, as measured by a Rating of Perceived Exertion (RPE), was greater for walking or running at the preferred transition speed (PTS). Twenty young, healthy adults (10 males, 10 females) walked on a treadmill at five speeds ranging from 70-110% of their individually measured PTS, and ran at five speeds ranging from 90-130% of their PTS while VO2 was monitored to determine each individual's energetically optimal transition speed (EOTS). Although the EOTS found during this study (2.24 m.s-1) was significantly greater than the PTS (2.06 m.s-1), RPE was significantly greater while walking at the PTS (13.5) than running at the PTS (10.0), suggesting that the gait transition during human locomotion does not take place in order to minimize metabolic energy consumption. PMID- 8231762 TI - Function of mono- and biarticular muscles in running. AB - In this study the function of leg muscles during stretch-shortening cycles in fast running (6 m.s-1) was investigated. For a single stance phase, kinematics, ground reaction forces, and EMG were recorded. First, rough estimates of muscle force, obtained by shifting the EMG curves +90 ms, were correlated with origin-to insertion velocity (VOI). Second, active state and internal muscle behavior were estimated by using a muscle model that was applied for soleus and gastrocnemius. High correlations were found between estimates of muscle force and VOI time curves for mono-articular hip, knee, and ankle extensor muscles. The correlation coefficients for biarticular muscles were low. The model results showed that active state of gastrocnemius was high during increase of origin-to-insertion length (LOI), whereas active state of soleus was low during the start of increase of LOI and rose to a plateau at the time lengthening ended and shortening started. It seems that the difference in stimulation between gastrocnemius and soleus is a compromise between minimizing energy dissipation and using the stretch-shortening cycle optimally. Furthermore, it was found that the net plantar flexion moment during running reached a value of 302 Nm, which was 158% and 127% higher than the peak values reached in maximal jump and sprint push offs, respectively. It was argued that the higher mechanical output in running than in jumping could be ascribed to the utilization of the stretch-shortening cycle in running. The higher values in running compared with sprinting, however, may lie in a difference in muscle stimulation. PMID- 8231763 TI - Self-selected exercise intensity of habitual walkers. AB - This study assessed self-selected exercise intensity of habitual walkers. Twenty nine healthy adults (22 females, 7 males; age (mean +/- SD) = 34.9 +/- 8.6 yr) performed a typical exercise walk while walking speed was measured by an unseen observer. On a subsequent occasion, the subjects walked at the same pace on a treadmill while several variables related to exercise intensity were measured. The mean self-selected walking pace was 1.78 +/- 0.19 m.s-1. Mean percents of VO2max and HRmax elicited by the treadmill exercise were 52 +/- 11% and 70 +/- 9%, respectively. Mean MET level was 5.2 +/- 1.2, and ratings of perceived exertion (RPE) averaged 10.9 +/- 1.6. Based on reported frequency and duration of walking, weekly energy expenditure in exercise walking was estimated to be 1127 +/- 783 kcal.wk-1. These data suggest that the self-selected exercise intensity of healthy, habitual exercise walkers meets the American College Sports Medicine's recommendation for improvement of cardiorespiratory fitness. These data further suggest that, in this population, the average weekly energy expended through walking reaches a level associated with improvements in health and longevity. PMID- 8231764 TI - The simplified V-slope method of detecting the gas exchange threshold. AB - A simplified V-slope method was used to visually determine the first point of departure from linearity of carbon dioxide output (VCO2) plotted against oxygen uptake (VO2). The point at which VCO2 departed from a line with a slope equal to 1.00 was visually selected as the gas exchange threshold during incremental exercise. The simplified method of threshold detection was compared with a computerized V-slope method. Both methods were used to determine the gas exchange threshold during incremental cycling (10 W.min-1) in 17 untrained female subjects. The thresholds occurred at 41.1% and 45.1% of VO2max using the computerized and simplified V-slope methods, respectively. The correlation between the oxygen uptake measured at the threshold using the two methods was 0.95. Power output, VO2, and heart rate values determined at the gas exchange threshold using the simplified V-slope method were significantly higher than the corresponding values obtained using the computerized V-slope method. The simplified V-slope method consistently placed the gas exchange threshold at the work rate that was about 10 W above the power output determined by the computerized method. Although the two methods were highly correlated, the simplified V-slope method tended to overestimate threshold values determined from a computerized gas exchange method. PMID- 8231765 TI - Validity of near infrared body composition analysis in children and adolescents. AB - The purpose of this study was to evaluate the validity and reliability of the Futrex 5000A near infrared (NIR) spectrophotometer for the assessment of body composition in children and adolescents. Forty-eight subjects (24 boys and 24 girls) with a mean age of 12.7 +/- 2.7 yr underwent three methods of body composition testing: NIR, densitometry by hydrostatic weighing (HYDRO), and skinfold anthropometry (SKF). Percent body fat (%BF) and fat free mass (FFM) derived from the Lohman's age-adjusted Siri equation served as the criterion. Within session test-retest reliability was determined for the NIR device for all subjects and between-week reliability was evaluated for all test methods in 14 subjects. Based on the excellent within- and between-session reliability (ICC ranged from 0.907-0.999), the system offers the potential of obtaining longitudinal data in growth and development studies. However, the significant mean differences, moderate correlations with the criterion (r = 0.62-0.71) and inflated standard errors of estimate (SEE = 4.9-5.5% BF, 2.2-2.9 kg FFM) and total prediction errors (TE = 5.5-8.0% BF, 2.7-3.7 kg FFM) indicate that refinement of prediction equations is needed to establish the measurement validity. Continued research with expanded populations is needed to further demonstrate and evaluate the utility of this device. PMID- 8231766 TI - Compendium of energy costs for various human activities. PMID- 8231767 TI - Modulation of IGF mRNA abundance during muscle denervation atrophy. AB - Changes in skeletal muscle activity cause dramatic alterations in muscle mass. Increased load on a muscle (synergistic overload) results in muscle hypertrophy. During hypertrophy, skeletal muscle concentrations of insulin-like growth factors (IGF-I and IGF-II) mRNAs increase. To clarify the role of IGFs in regulating muscle mass, this study examined whether IGF-I and -II mRNA levels were altered during decreased muscle activity (denervation). Gastrocnemius weights decreased 4.2%, 7.7%, 18.1%, 27.7%, 35.1%, 45.0%, and 60.3% at 2, 3, 5, 7, 10, 12, and 17 d following denervation, respectively. Muscle DNA content remained constant throughout the first 12 d after surgery, but increased above control levels at day 17. During the first week after surgery, gastrocnemius IGF-II mRNA remained constant. However, IGF-II mRNA abundance was 2.5-fold greater than controls by 10 d of denervation, 3-fold by 12 d, and 6.8-fold by 17 d. On the other hand, IGF-I mRNA levels were not affected by denervation. In conclusion, although increased muscle activity results in a change of IGF-I mRNA expression, decreased muscle activity has no effect on IGF-I mRNA expression. In contrast, IGF-II mRNA levels increase with long-term denervation as well as with increased muscle activity. This study suggests that muscle activity may not be the only factor affecting IGF I and -II expression. PMID- 8231768 TI - Carbohydrate dependence during marathon running. AB - To test the hypothesis that marathon running is dependent on lipid oxidation, 12 post-absorptive males (31.9 +/- 2.1 yr) ran a treadmill marathon and substrate utilization was assessed. Subjects were placed into a fast (F < or = 2 hr, 45 min; 73.3% VO2max), or a slow (S < or = 3 hr, 45 min; 64.5% VO2max) marathon group. The day before testing subjects rested, but ate their normal diet. Subjects were tested in the morning after an overnight fast, and only tap water, at a rate of 1 l.h, was ingested during exercise. Blood glucose concentration rose at exercise onset, peaked at approximately an hour, but then decreased over time remaining at or above resting levels. Free fatty acids and glycerol rose continuously. No significant differences in plasma FFA, glycerol, or blood glucose concentrations were observed between F or S groups during the marathon. Mean blood lactate concentration was significantly higher (P < 0.05) in the F (2.1 +/- 0.3 mM) group than the S (1.2 +/- 0.2 mM) during exercise. Mean plasma epinephrine was significantly higher in the F (0.9 +/- 0.2 ng.ml-1) than the S (0.6 +/- 0.2 ng.ml-1) group; norepinephrine was also higher in F (3.9 +/- 1.4 ng.ml-1) than the S (2.5 +/- 0.9 ng.ml-1, P < or = 0.05). Blood lactate and epinephrine concentrations correlated significantly (4r = 0.76 and 0.78 in F and S groups, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231769 TI - Effects of nicotinic acid on respiratory exchange ratio and substrate levels during exercise. AB - The purpose of this study was to investigate the effects of pharmacologic treatment with nicotinic acid (NA) (3 g NA.d-1) on fuel utilization during exercise and on total and HDL-cholesterol levels of trained runners. Eight experienced male runners performed four 30-min submaximal treadmill runs (SUB) at approximately 60% of aerobic capacity: the first served as the control condition, whereas SUBs 2, 3, and 4 were conducted at the onset, midpoint, and conclusion of the 3-wk NA treatment period. A 1-g NA dose was ingested 1 h prior to SUBs 2-4, which were conducted in the morning following a 12-h fast. The respiratory exchange ratio (RER) was significantly higher during SUBs 2, 3, and 4 (0.919 +/- 0.009, 0.898 +/- 0.007, and 0.896 +/- 0.009, respectively) than during SUB1 (0.871 +/- 0.008); and the RER during SUB2 was also significantly higher than that during SUB3 and SUB4 (P < 0.001). Serum free fatty acid (FFA) and glycerol levels were significantly lower during SUBs 2, 3, and 4 as compared with SUB1 (P < 0.001). Total cholesterol showed a significant decrease (from 195.3 +/- 9.2 to 174.5 +/- 9.2 mg.dl-1) and HDL cholesterol showed a significant elevation (from 56.2 +/- 2.9 to 63.0 +/- 3.9 mg.dl-1) during the NA treatment period (P < 0.05). The RER, FFA, and glycerol data demonstrate a persistent inhibition of fat utilization during submaximal exercise due to NA treatment. The 3-wk NA treatment period also proved to be sufficient to effect changes in total and HDL cholesterol levels of these runners. PMID- 8231770 TI - A diminished aortic-cardiac reflex during hypotension in aerobically fit young men. AB - We compared the aortic-cardiac baroreflex sensitivity in eight average fit (AF: VO2max = 44.7 +/- 1.3 ml.kg-1 x min-1) and seven high fit (HF: VO2max = 64.1 +/- 1.7 ml.min-1 x kg-1) healthy young men during hypotension elicited by steady state sodium nitroprusside (SN) infusion. During SN mean arterial pressure (MAP) was similarly decreased in AF (-12.6 +/- 1.0 mm Hg) and HF (-12.1 +/- 1.1 mm Hg). However, the increases in heart rate (HR) were less (P < 0.023) in HF (15 +/- 3 bpm) than AF (25 +/- 1 bpm). When sustained neck suction (NS, -22 +/- 1 torr in AF and -20 +/- 1 torr in HF, P > 0.05) was applied to counteract the decreased carotid sinus transmural pressure during SN, thereby isolating the aortic baroreceptors, the increased HR remained less (P < 0.021) in HF (8 +/- 2 bpm) than AF (16 +/- 2 bpm). During both SN infusion and SN+NS, the calculated gains (i.e., delta HR/delta MAP) were significantly greater in AF (2.1 +/- 0.3 and 1.3 +/- 0.2 bpm.mm Hg-1) than HF (1.2 +/- 0.2 and 0.6 +/- 0.2 bpm.mm Hg-1). However, the estimated carotid-cardiac baroreflex sensitivity (i.e., the gain difference between the stage SN and SN + NS) was not different between AF (0.7 +/- 0.2 bpm.mm Hg-1) and HF (0.6 +/- 0.1 bpm.mm Hg-1). These data indicated that the aortic-cardiac baroreflex sensitivity during hypotension was significantly diminished with endurance exercise training. PMID- 8231771 TI - Effects of body mass on exercise efficiency and VO2 during steady-state cycling. AB - Oxygen uptake (VO2) and exercise efficiency during cycle ergometer exercise are considered to be independent of body mass. To determine the validity of this assumption, 50 females ranging in body mass from 41.5-98.9 kg exercised on a cycle ergometer with no load at 60 rpm and at 25, 50, 75, and 100 W at 60 and 90 rpm. Gross VO2 and efficiency, net VO2 and efficiency, work VO2 and efficiency, and delta efficiency were computed. Gross and net VO2 were significantly and positively correlated with body mass at all work rates and pedal frequencies. Gross efficiency was significantly and negatively correlated with body mass at all work rates and pedal frequencies. Work VO2 and body mass were not significantly correlated. The correlations between work and delta efficiency and body mass were not significant. Since body mass was found to be significantly correlated with gross VO2, the following equation was developed using stepwise multiple regression to predict gross VO2: VO2 (ml.min-1) = 10.9 (work rate, W) + 8.2 (pedal rate, rpm) + 8.3 (body mass, kg) - 559.6. These data suggest that body mass should be considered when estimating the oxygen uptake during cycle ergometer exercise. PMID- 8231772 TI - Effect of preload and range of motion on isokinetic torque in women. AB - The purposes of this study were to compare the effects of two preload settings (0 N, 75 N) and two ranges of motion (5-90 degrees ROM, 25-70 degrees ROM) on torque output of the knee musculature. Twenty females were randomly assessed for isokinetic concentric (CON) and eccentric (ECC) torque of the knee extensor (EXT) and flexor (FLEX) musculature at a velocity of 1.62 rads.s-1. Two four-way ANOVAs (muscle x mode x range of motion x preload) revealed significant differences in average torque between the preload and range of motion conditions (CONEXT: 5-90 degrees ROM, 75 N = 74.3 +/- 17.2 Nm; 5-90 degrees ROM, 0 N = 68.3 +/- 17.2 Nm; 25-70 degrees ROM, 75 N = 79.0 +/- 13.0 Nm; 25-70 degrees ROM, 0 N = 71.5 +/- 20.8 Nm) (ECCEXT: 5-90 degrees ROM, 75 N = 85.6 +/- 28.6 Nm; 5-90 degrees ROM, 0 N = 82.8 +/- 27.8 Nm; 25-70 degrees ROM, 75 N = 97.7 +/- 23.4 Nm; 25-70 degrees ROM, 0 N = 93.6 +/- 26.5 Nm) (CONFLEX: 5-90 degrees ROM, 75 N = 43.5 +/- 9.2 Nm; 5-90 degrees ROM, 0 N = 43.1 +/- 5.6 Nm; 25-70 degrees ROM, 75 N = 44.2 +/- 8.9 Nm; 25-70 degrees ROM, 0 N = 41.2 +/- 8.9 Nm) (ECCFLEX: 5-90 degrees ROM, 75 N = 56.7 +/- 16.3 Nm; 5-90 degrees ROM, 0 N = 55.6 +/- 17.8 Nm; 25-70 degrees ROM, 75 N = 57.3 +/- 14.0 Nm; 25-70 degrees ROM, 0 N = 51.8 +/- 14.0 Nm) (P < 0.05). No differences in peak torque values were observed. Based on the findings of this study, preload and range of motion should remain constant between and among subjects if average torque is used as a criterion measure. PMID- 8231773 TI - State anxiety and blood pressure following 30 min of leg ergometry or weight training. AB - The effect of different modes of acute exercise on state anxiety and blood pressure was examined. Collegiate varsity athletes (11 females and 15 males) completed 30-min sessions of leg cycle ergometry or weight training in a randomized order on separate days. Both exercise modes were performed within 70 80% of the maximum capacity of each subject. State anxiety, systolic (SBP) and diastolic (DBP) blood pressure were measured at baseline prior to exercise, immediately post-exercise, 20 min and 60 min following exercise. Repeated measures ANOVA detected significant (P < 0.05) trial by condition effects for state anxiety and SBP. State anxiety increased (P < 0.001) immediately following weight training but returned to baseline for the remaining assessments. State anxiety decreased (P < 0.05) below baseline at 60 min following ergometry. SBP did not decrease following weight training but was reduced (P < 0.05) by 6.5 mm Hg below baseline at 60 min following ergometry. DBP did not change following either exercise mode. In summary, state anxiety and blood pressure responses to acute physical activity appear to be dependent on the exercise mode, with reductions in state anxiety and systolic blood pressure being associated with ergometry but not weight training. PMID- 8231774 TI - The Social Physique Anxiety Scale: construct validity in adolescent females. AB - Hart, Leary, and Rejeski have developed the Social Physique Anxiety Scale (SPA), a measure of the anxiety experienced in response to having one's physique evaluated by other people. The present study cross-validated the psychometric properties of this measure in a sample (N = 236) of adolescent competitive female gymnasts. Employing structural equation modeling, the proposed unidimensional factor structure of the SPA was supported, although some questions regarding the robustness of the fit are raised. Construct validity was demonstrated by significant inverse relationships between aspects of physical efficacy (perceived physical ability and physical self-presentation confidence) and degree of social physique anxiety. These findings are discussed in terms of possible alternative factor structures and integration of social anxiety and other psychosocial constructs to better understand physical activity behavior. PMID- 8231775 TI - Effects of electrical stimulation and upper body training after spinal cord injury. AB - The purpose of this study was to measure the cardiorespiratory improvements during the initial weeks of training in individuals with spinal cord injury (SCI). Eight adult volunteers (ages 23-41) with paraplegia (T4-L1, N = 7) and low levels of quadriplegia (C7, N = 1) participated in a 12-wk training program consisting of 6 wk of computerized functional electrical stimulation leg ergometry (CFES LE) followed by 6 wk of combined arm ergometry and CFES LE (hybrid ergometry; HE). Measures of peak oxygen consumption (pV02) and immediate post-exercise blood lactate (La-) were collected during an intermittent CFES LE graded exercise test (3-min stages; 1/8-kp increments) prior to training (0T), following 6 wk of CFES LE training (6T) and following 6 wk of HE training (12T). Voluntary arm ergometry (AE) graded exercise tests were also completed at each of 0T, 6T, and 12T, and an intermittent graded HE test was also given at 6T and 12T. Following the 6 wk of CFES LE, AE pVO2 increased from 1.14 +/- 0.09 l.min-1 to 1.39 +/- 0.172 l.min-1 (P < 0.05), and CFES LE pVO2 increased from 0.51 +/- 0.05 l.min-1 to 0.83 +/- 0.06 l.min-1 (P < 0.05). Following the 6 wk of HE, HE pVO2 increased from 1.31 +/- 0.15 l.min-1 to 1.49 +/- 0.14 l.min-1 (P < 0.05). HE pVO2 was 58%-60% higher than CFES LE pVO2 at the mid- and post-testing periods. HE pVO2 was 14% higher than AE pVO2 at the post-testing period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231776 TI - Using maximal and submaximal aerobic variables to monitor elite cyclists during a season. AB - The purpose of this study was to identify sensitive physiological indicators for monitoring the progress in training state of elite cyclists throughout a training and competition season. Seven elite male cyclists performed maximal oxygen consumption (VO2max) tests and submaximal tests on three or four different occasions during their training and competition season. The submaximal test consisted of three successive 16-min bouts at 150, 200, and 250 W interspersed with rest periods between each load. Pedalling rate varied progressively from 50 120 rpm. Results showed that maximal heart rate (HR), ventilation (Ve), gross mechanical efficiency (MEG), and VO2max did not change during the season. Moreover, ventilation threshold (VT) did not change during the season, nor did HR, Ve, and VO2 at VT. However, changes were found in physiological variables during the submaximal test. HR decreased significantly during the season, as did VO2 (P < 0.05) for a standardized workload. On the other hand, at the higher power outputs (200 W and 250 W) MEG increased during the season. These results suggest that VO2max may not be a good indicator of enhanced capacity in elite cyclists. Rather, a standardized efficiency test during which submaximal variables such as HR, VO2, and MEG are monitored might be a more sensitive indicator of the progress in the training state of elite male cyclists during a training and competition season. PMID- 8231777 TI - Cross-validation of the Slaughter skinfold equations for children and adolescents. AB - Prior to sexual maturation, children and adolescents have more water and less bone mineral content than adults, resulting in less dense fat-free body mass (FFM). This suggests that previously established adult skinfold/density equations are inappropriate for use with children and adolescents for the prediction of body fatness (%BF) and FFM. To overcome this problem, Slaughter and colleagues have introduced new skinfold (SKF) equations that take into account the changing density of FFM in children and adolescents as they mature. The purpose of our study was to cross-validate a select set of the Slaughter SKF equations by comparing them with a criterion measurement (Lohman's Siri age-adjusted body density equation) in 122 subjects ranging in sexual maturation from pre- to post pubescent and ranging in age from 8-17. Our cross-validation found very high intraclass (reliability) correlations (ICCs = 0.98-0.99) and high validity correlations (rs = 0.79-0.99). The standard errors of the estimate for %BF ranged from 3.5-4.6% and total errors for %BF ranged from 3.6-4.6%. The Slaughter equation using tricep and calf SKF for females was significantly different (P < 0.05) from the criterion measure in its prediction of %BF. In males, there was an interaction between the SKF equation and subject maturation level. The data indicate that the Slaughter SKF equations hold promise for estimating body composition in children and adolescents but are still in need of refinement. PMID- 8231778 TI - Predicting max HR and the HR-VO2 relationship for exercise prescription in obesity. AB - This research derived regression equations for predicting maximal heart rate (MHR) and examined the relationship between relative oxygen consumption (VO2) and heart rate (HR) in obese (N = 86, body fat > 30%, hydrostatic weighing) compared with normal-weight (N = 51, body fat < or = 30%) adults. Simultaneous measurements of HR and VO2 were recorded at rest and every minute during a maximal graded exercise test. When MHR was regressed on age, two distinct equations for the obese and normalweights were generated. The relationship between %MHR and %max VO2 was similar between groups (r = 0.83, obese; r = 0.87 normalweights). Likewise, when %max VO2 was regressed on %max heart rate range similar equations were derived fro the obese (r = 0.81) and normalweights (r = 0.84). Correlation between Karvonen's predicted HR at a submaximal VO2 and the true HR at that VO2 was 0.88, regardless of adiposity. These data indicate that when predicting MHR in normalweights the equation 220-Age can be used, but for obese individuals the equation 200-0.5 x Age is more accurate; each having 12 as a standard error of estimate. Once MHR is determined, either the straight percentage technique or Karvonen's method would be appropriate for prescribing exercise intensity for both populations. PMID- 8231779 TI - Redefining cervical spinal stenosis using MRI. PMID- 8231780 TI - Electroencephalographic measures of attentional patterns prior to golf putt. PMID- 8231781 TI - Athletes, athletics, and sudden cardiac death. AB - The pathological causes of sudden death during athletics varies with the age of the competitor. Congenital abnormalities are the predominant cause of exercise related deaths in subjects under age 30 yr whereas atherosclerotic coronary artery disease is the primary cause of such deaths in adults. Cardiovascular screening programs designed to reduce the incidence of exercise deaths are limited by the rarity of exercise events, the poor predictive value of the tests, and the cost of testing. Nevertheless, we recommend that young athletes be examined to detect conditions associated with exercise complications and that new symptoms in young and old athletes be carefully evaluated. PMID- 8231782 TI - Mediopatellar synovial plica of the knee in athletes: results of arthroscopic treatment. AB - The long-term results (9-87 months) of the arthroscopic treatment of unilateral mediopatellar synovial plica of the knee in 102 athletes are reported. In all patients a large and/or fibrotic inflamed mediopatellar plica impinging on the femoral condyle on knee flexion was identified and trimmed. Chondromalacia patellae was detected in 38 patients. The impingement lesion was seen in 13 knees. In one patient, a minor tear of the lateral meniscus was trimmed, and one patient was found to suffer from a clinically asymptomatic partial tear of the anterior cruciate ligament. Isometric strength testing of the quadriceps revealed marked decrease at the second postoperative week, with subsequent gradual recovery. The results obtained justify arthroscopic treatment in a young, extremely active population. PMID- 8231783 TI - Effects of patella taping on patella position and perceived pain. AB - Anterior knee pain syndrome (AKPS) represents a significant challenge for the sports medicine clinician. One proposed etiological factor for AKPS is poor vastus medialis obliquus (VMO) control, resulting in lateral glide of the patella. Patella taping has been advocated to increase VMO control. The purpose of this study was to determine the effects of patella taping on patella position and perceived pain. Twelve subjects (age = 29 +/- 9 yr; weight = 70.9 kg +/- 17.8; height = 174.0 cm +/- 8.1) with AKPS currently using patella taping procedures with a decrease in their symptoms participated. Each subject had Merchant's view x-rays taken pre- and post-taping while performing an isometric quadriceps contraction to determine patella rotation and patella congruency angles. Subjects also completed a visual analog pain scale after performing a 0.2 m (8") step-down both pre- and post-taping. Paired t-tests revealed no significant change in patellofemoral congruency (P = 0.98) and patella rotation angles (P = 0.80). Significant reduction (50%) in subject pain level was revealed by the visual analog scale (t(15) = 4.99; P < 0.0005). Results demonstrate that patella taping significantly reduced the perceived pain levels during a 0.2-m step-down; however, this reduction in pain was not associated with patella position changes. PMID- 8231784 TI - Effects of maternal aerobic fitness on cardiorespiratory responses to exercise. AB - We hypothesized that aerobically fit women who continued to exercise throughout pregnancy would have enhanced cardiorespiratory responses to exercise. Physically active (N = 10) and sedentary (N = 6) subjects were compared during steady-state (15 min) semi-recumbent cycle exercise performed at a given heart rate (HR; 140 b.min-1) twice during pregnancy (25 wk, 36 wk) and 12 wk postpartum. Indirect calorimetry was used to measure volumes and fractional concentrations of expired gases. Cardiac output was estimated via CO2 rebreathing. Data were analyzed with repeated measures ANOVA. Caloric expenditure during exercise was significantly (P < 0.001) greater in the physically active (7.2 kcal.min-1) compared with sedentary (4.7 kcal.min-1) subjects. Alveolar ventilation and cardiac output responses to exercise were proportionally greater (P < 0.001) in the aerobically fit subjects. Ventilatory equivalents for O2 and physiological dead space/tidal volume ratios were significantly (P < 0.01) lower in the physically active subjects during exercise. In contrast, ratings of perceived exertion during exercise did not differ between subject groups. It appears that a physically active woman's enhanced cardiorespiratory responses to acute exercise are maintained during pregnancy if she continues her aerobic fitness program throughout gestation. PMID- 8231785 TI - Contractile properties of skinned fibers from hypertrophied skeletal muscle. AB - We have previously shown deficits in specific tension (N.cm-2) in whole hypertrophied skeletal muscle (18-30%). The purpose of the present study was to determine if this intrinsic deficit exists in chemically skinned, hypertrophied single fibers, and also to measure their Ca2+ sensitivity. One group of rats (N = 4) had bilateral surgical ablation of the gastrocnemius and soleus muscles, and another group (N = 4) had the gastrocnemius and plantaris muscles ablated to induce hypertrophy in the plantaris and soleus muscles, respectively. Thirty days after surgery five fibers were dissected and analyzed from each of eight muscles per group, including control muscles. In hypertrophied plantaris and soleus muscles, fiber cross-sectional area (CSA) was 20% and 29% greater, respectively, than control values. Maximal Ca2+ activated tension (Po, pCa 4.5) was 15% greater in hypertrophied plantaris fibers and 18% greater in hypertrophied soleus fibers compared with respective control values (P < 0.05). Therefore, Po expressed per fiber CSA was slightly depressed (3-8%) in both plantaris and soleus fibers (P < 0.05). Regarding Ca2+ sensitivity, the Ca2+ concentration to elicit 50% of Po (i.e., pCa50) was significantly lower in hypertrophied soleus (58%) and plantaris (29%) fibers. This leftward shift in the force-pCa curve reflects greater Ca2+ sensitivity in hypertrophied fibers. Since the single fiber Po/CSA was only slightly decreased, these data suggest that during tetanic stimulation of whole muscle Ca2+ delivery to the contractile apparatus may be impaired.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231786 TI - Pontine and extrapontine myelinolysis: a neurologic disorder following rapid correction of hyponatremia. AB - Neurologic disorders developing after correction of severe, symptomatic hyponatremia were studied in 14 patients. None had a hypoxic event or other identifiable cause for the neurologic illness. Neurologic deterioration began about 3 days after correction and often followed a period of improvement in hyponatremic encephalopathy. Although the symptoms were as mild as transient confusion in 1 patient, they were more severe in the others. Typically, spastic quadriparesis, pseudobulbar palsy, and impairment in the level of consciousness progressed for up to 7 days. Improvement generally began 2 weeks after correction and continued for up to a year in some patients. Routine spinal fluid analysis was usually normal, but myelin basic protein concentration was elevated in all patients in whom it was measured. Electroencephalograms commonly showed nonfocal slowing. Brainstem auditory evoked potential latencies were prolonged in some patients. Brain imaging was normal in the initial week of illness, while later scans, obtained in 9 patients, showed central pontine and/or symmetric extrapontine lesions. The clinical manifestations and distribution of lesions seen on imaging demonstrate that neurologic illness following correction of hyponatremia is due to myelinolysis. Although this neurologic disorder typically followed an elevation in serum sodium > 18 mEq/L/24 hr, it sometimes followed a rise as slow as 10 mEq/L/24 hr and 21 mEq/L/48 hr. Whenever possible, the rate of correction of hyponatremia should be kept below these values in order to minimize the risk of myelinolysis. PMID- 8231787 TI - Meningococcal disease in patients with late complement component deficiency: studies in the U.S.S.R. AB - The purpose of this study was to examine the occurrence of late complement component deficiency (LCCD) states in the USSR. Thirty deficient individuals were detected: 27 with C8 beta and 3 with C7 deficiency. Among individuals with a first episode of meningococcal infection, about 1% had LCCD, whereas among patients with recurrent bacterial meningitis the prevalence of LCCD rose to approximately 50%. This corresponds to a prevalence for LCCD of approximately 12 per 100,000 in the general population. The individuals with LCCD identified in this study experienced about 77 episodes of meningococcal disease and acute bacterial meningitis. Mathematical analysis of the morbidity from meningococcal disease in individuals with LCCD demonstrated that the probability of disease did not change with the age of the patient and was not affected by prior episodes of infection. This finding suggest that in contrast to the situation in the general population, prior infection fails to protect the deficient individual from recurrent disease. In comparison to complement-sufficient persons, the course of disease in individuals with LCCD is less severe, as shown by a reduction in the number of episodes of endotoxic shock and mortality as well as their more rapid recovery. These findings suggest that exuberant complement activation and concomitant formation of membrane attack complexes during meningococcal infection in complement-sufficient patients plays an important role in the activation and injury of peripheral blood cells and endothelial cells during endotoxic shock. PMID- 8231788 TI - Bloom syndrome: a mendelian prototype of somatic mutational disease. AB - Spontaneous mutations in human somatic cells occur far more often than normal in individuals with Bloom syndrome. The basis for understanding these mutations and their developmental consequences emerges from examination of BS at the molecular, cellular, and clinical levels. The major clinical feature of BS, proportional dwarfism, as well as its major clinical complication, an exceptionally early emergence of neoplasia of the types and sites that affect the general population, are attributable to the excessive occurrence of mutations in somatic cells. Here, the following aspects of BS are discussed: (i) the BS phenotype; (ii) neoplasia in BS, including the means--the Bloom's Syndrome Registry--by which the significant risk for diverse sites and types of cancer in these patients was revealed; (iii) the biological basis for the cancer proneness of BS; and, finally, (iv) the significance for both basic human biology and clinical medicine of BS as the prototype of somatic mutational disease. PMID- 8231789 TI - Subaortic stenosis: a reviewed concept of the disease. 1959. PMID- 8231790 TI - Subaortic stenosis revisited: the importance of the dynamic pressure gradient. AB - Brachfeld and Gorlin's revised concept of subaortic stenosis derived from their recognition that the large magnitudes of the pressure gradients were incompatible with the relatively mild anatomic lesions found at surgery in 3 of their patients, and the rapidity of their arterial pulse upstroke. They proposed that a gradient caused by a superimposed systolic stenosis was responsible for the large pressure gradients and the overestimation of the severity of the discrete subaortic stenosis (DSAS). A fourth patient had no anatomic cause for the pressure gradient, and findings compatible with hypertrophic cardiomyopathy (HCM). All 4 patients had septal hypertrophy which they felt was instrumental in the brisk pulse contour and dynamic gradients across the outflow tract. In the ensuing decades, imaging techniques have been developed which permit detailed studies of ventricular ejection patterns in dynamic gradients associated with HCM and DSAS. These studies have been interpreted variously. The prevailing view is that there is a dynamic obstruction that increases progressively in severity during systole, as proposed by Brachfeld and Gorlin. An opposing view is that dynamic gradients can occur in the absence of any hindrance to ejection, and that these gradients instead result from rapid and complete emptying of the ventricle. Regardless of their cause, dynamic gradients are often superimposed upon gradients caused by DSAS and valvar aortic stenosis, leading to exaggerated estimates of severity. These dynamic gradients are uncovered when the anatomic cause of stenosis is removed, and seemingly increase the postoperative morbidity. A greater understanding of the significance of dynamic gradients and the mechanism(s) responsible for them should lead to more rational management of DSAS and HCM in the future. PMID- 8231791 TI - [Organizational and social factors on performance among managers of the occupational health service (results of an opinion survey)]. AB - This paper presents results of a questionnaire survey carried out among managers of the occupational health service (OHS). The main objective of the survey was to evaluate organisational and social conditions of work and to identify what is needed to improve managerial qualifications. The study provided information on managers' attitude towards goals and responsibilities of institutions run by them, the efficiency of decision-making process, proposed changes in organizational structures and man power management. The results of the study confirmed the need for improving postgraduate training designed for OHS managers. PMID- 8231792 TI - [Contact allergy caused by formaldehyde, based on materials from the Institute for medical occupations in Lodz]. AB - Incidence of formaldehyde allergy was analysed in 3,705 subjects with dermatitis and in 1,530 subjects with allergic dermatitis examined within the last fifteen years. Rates of increase or decrease in relative risk during that period was analysed in women, in men and in the whole group. In the whole material no differences in the incidence of allergy according to sex were found. Among women allergic to formaldehyde, dermatitis was found in 4.5 per cent and allergic dermatitis in 11.6 per cent. Among men the per cent was equal to 4.6 and 10.5, respectively. A significant upward tendency was found in the risk of formaldehyde allergy in men. The main sources of occupational allergies were: disinfectant agents, end-use finish and cloth dyes, cutting fluids and plastics. PMID- 8231793 TI - [Health threat from asbestos dust at certain railroad workplaces. I. Evaluation of the work environment]. AB - Dust levels and respirable asbestos fibre concentrations were measured at workposts of wiremen and mechanics involved in repairs of railway engines. Measurements and data obtained revealed an excess of allowable concentrations of asbestos fibres in a number of workposts under study. Conclusions and recommendations concerning the work organization and workers' health protection are presented. PMID- 8231794 TI - [Evolution of radiologic changes in lungs of workers occupationally exposed to asbestos dust]. AB - The dynamic of radiological changes in lungs of 458 workers exposed to chrysotile asbestos dust and in 65 retired workers previously exposed to the same dust was assessed. There was a 5-year interval between initial and control examinations in the group of acting workers and an 8-year interval in the group of retired workers. The control examination of workers with employment below 10 years did not reveal radiological changes which could indicate asbestosis. In the group of 400 workers with employment over 10 years new cases of asbestosis were diagnosed in 34 persons (8.5%) during the control examination. During the control examination progress of pleural changes was observed in 4% of males and 3.8% of females. As the period of exposure to asbestos had expanded the occurrence of small, irregular shadows, especially in persons working longer than 15 years, was more often noticed. The aforesaid aberrations have progressed in 11.7% of males and 9.5% of females when compared with the results of the initial examination. In the group of retired workers 15 (23.1%) new cases of asbestosis were detected 8 years after the termination of occupational exposure. Small, irregular shadows, limited in number as well as pleural changes have progressed in approximately 22% of retired males and females. PMID- 8231795 TI - [Healthy choice of occupation for juveniles in the textile and clothing industry in light of preventive examinations]. AB - Choice of profession in line with natural liking or aptitude for a certain type of work and physical fitness is considered as a proper one. The aim of this study was to verify the effectiveness of pre-placement examinations of juvenile leaving primary schools. The study covered: (1) 210 students of textile and clothing schools; (2) 70 students from special care classes of the textile school; and (3) 312 graduates from primary schools applying for admission to textile and clothing schools. The study was carried out at the Nofer Institute of Occupational Medicine, Lodz, Poland. The reported subjective symptoms were usually limited to the headache (27%). The most prevalent abnormalities were visual organ impairments. Within the group of special care students from the textile school, scoliosis and functional disturbances of the nervous system were detected. The study revealed that both the range of pre-placement examinations of juvenile and the recognition of the criteria for certifying work capability are inadequate. PMID- 8231796 TI - [Work disability in certain occupations among workers in the electronics industry]. AB - Occupational and working environment factors have always occupied an important place in studies on sickness absenteeism. The study performed during the years 1987-1988 in two electronic industry plants involved such elements of the working environment as the place of a worker in the production process and the occupation performed. Our analysis was aimed at indicating differences in the temporary work disability, in particular due to some groups of diseases, between occupations essential for the manufacturing process. The frequency of sickness absenteeism in general and due to main groups of diseases expressed in terms of percentage of sick persons with temporary work disability was analysed. The Chi 2 test was used to determine the relationship between individual groups. The analysis of sickness absenteeism in individual groups was performed according to sex and age. The analysis allowed to indicate groups of workers with increased risk of sickness absenteeism in the plants under study. They are mostly workers involved in the basic production and in chemical processes. Sickness absenteeism of those workers was higher than the average by 26% in Plant I and by 33% in Plant II. The highest sickness absenteeism rate for that group was due to diseases of the nervous system--71%; the gastrointestinal system--46%; and the musculo-skeletal system- 81%. The relationship between the occurrence of sickness absenteeism and the type of activity was statistically significant (p < 0.05) in both Plants but in relation to selected groups of occupation it was significant only in Plant I. PMID- 8231797 TI - [Traffic accidents in the midst of busy municipal transportation in Lodz]. AB - Records of 1,543 municipal transportation accidents registered at the Department of Inspection, Municipal Transportation Enterprise, Lodz were the subject of analysis presented in this paper. The paper includes data on kinds of road accidents which took place in 1991, causes of accidents and the impact of bus and tram drivers on their occurrence. The occurrence of road accidents in relation to individual parts of a year, a week or a day was also analysed. The study provides a detailed description of traffic accidents which happened in the Lodz municipal transportation in 1991 with special reference to human factors. PMID- 8231798 TI - [Organizing evaluation of chemical substances in EEC countries and in Poland prior to their admission to trade turnover]. AB - This paper presents systems how chemical substances, those in use and new ones, are evaluated in the EEC countries and in Poland prior to their admission to trade turnover. The Polish hygienic monitoring of chemical substances in comparison with systems operating in the Western countries requires a very thorough verification. New organizational concepts of evaluation of chemical substances before their admission to trade turnover in Poland are generally outlined. It is stressed that issues related to production and entering the market with chemical substances require new legal regulations. It also is necessary to set up a national centre responsible for implementing provisions of the Chemical Substances Act which is now being drafted. PMID- 8231799 TI - [Biological effect of wood dust]. AB - The biological effect of exposure to wood dust depends on its composition and the content of microorganisms which are an inherent element of the dust. The irritant and allergic effects of wood dust have been recognised for a long time. The allergic effect is caused by the wood dust of subtropical trees, e.g. western red cedar (Thuja plicata), redwood (Sequoia sempervirens), obeche (Triplochiton scleroxylon), cocabolla (Dalbergia retusa) and others. Trees growing in the European climate such as: larch (Larix), walnut (Juglans regia), oak (Quercus), beech (Fagus), pine (Pinus) cause a little less pronounced allergic effect. Occupational exposure to irritative or allergic wood dust may lead to bronchial asthma, rhinitis, alveolitis allergica, DDTS (Organic dust toxic syndrome), bronchitis, allergic dermatitis, conjunctivitis. An increased risk of adenocarcinoma of the sinonasal cavity is an important and serious problem associated with occupational exposure to wood dust. Adenocarcinoma constitutes about half of the total number of cancers induced by wood dust. An increased incidence of the squamous cell cancers can also be observed. The highest risk of cancer applies to workers of the furniture industry, particularly those dealing with machine wood processing, cabinet making and carpentry. The cancer of the upper respiratory tract develops after exposure to many kinds of wood dust. However, the wood dust of oak and beech seems to be most carcinogenic. It is assumed that exposure to wood dust can cause an increased incidence of other cancers, especially lung cancer and Hodgkin's disease. The adverse effects of microorganisms, mainly mould fungi and their metabolic products are manifested by alveolitis allergica and ODTS. These microorganisms can induce aspergillomycosis, bronchial asthma, rhinitis and allergic dermatitis. PMID- 8231800 TI - [Air ions and their influence on live organisms]. AB - This work discusses the influence of air-ions on human and animal organisms. It presents problem of measurements of airs-ions parameters and gives suggestions how to improve methods of investigation in order to analyse that influence in a more profound way. PMID- 8231801 TI - The filamentous haemagglutinin, a multifaceted adhesion produced by virulent Bordetella spp. AB - Filamentous haemagglutinin (FHA) is the major attachment factor produced by virulent Bordetella spp. Similar to the other virulence factors, its production is tightly regulated by a two-component system in response to environmental changes. Although of impressive size (c. 220 kDa), it is very efficiently released into the culture supernatant of Bordetella pertussis. Its biogenesis involves complex processing of a larger precursor with a calculated molecular mass of 370 kDa. Export of FHA into the culture medium depends on an outer membrane protein homologous to haemolysin accessory proteins. Purified extracellular FHA is able to increase the adherence of other pathogens to the host, which may contribute to super-infection in whooping cough. Although FHA- mutants colonize lungs as efficiently as the wild-type parent strains, immune responses against FHA appear to protect against colonization. Unlike many other adhesins, FHA expresses at least three different attachment activities, one specific for the CR3 integrins of macrophages, one involving a carbohydrate binding site, specific for interactions with cilia, and a heparin-binding activity that may be important for interaction of B. pertussis with epithelial cells or extracellular matrices. PMID- 8231802 TI - The interaction between coumarin drugs and DNA gyrase. AB - The coumarin group of antibiotics have as their target the bacterial enzyme DNA gyrase. The drugs bind to the B subunit of gyrase and inhibit DNA supercoiling by blocking the ATPase activity. Recent data show that the binding site for the drugs lies within the N-terminal part of the B protein, and individual amino acids involved in coumarin interaction are being identified. The mode of inhibition of the gyrase ATPase reaction by coumarins is unlikely to be simple competitive inhibition, and the drugs may act by stabilizing a conformation of the enzyme with low affinity for ATP. PMID- 8231803 TI - The partition (par) locus of pSC101 is an enhancer of plasmid incompatibility. AB - The incompatibility that pSC101-derived plasmids express toward each other is mediated by directly repeated sequences (iterons) located near the plasmid's replication origin. We report here that the pSC101 par locus, which stabilizes plasmid inheritance in dividing cell populations and alters DNA superhelicity, can function as a cis-acting enhancer of incompatibility, which we show is determined jointly by the copy number of the plasmid and the number of iterons per copy. A single synthetic 32 bp iteron sequence carried by the pUC19 plasmid confers strong pSC101-specific incompatibility in the absence of any other pSC101 sites but requires the par locus to express strong incompatibility when carried by a lower-copy-number plasmid. We propose a model by which the par locus can enhance the apparently antagonistic processes of incompatibility and pSC101 DNA replication while concurrently facilitating plasmid distribution during cell division. PMID- 8231804 TI - AmpG, a signal transducer in chromosomal beta-lactamase induction. AB - The chromosomal ampC beta-lactamase in Citrobacter freundii and Enterobacter cloacae is inducible by beta-lactam antibiotics. When an inducible ampC gene is introduced on a plasmid into Escherichia coli together with its transcriptional regulator ampR, the plasmid-borne beta-lactamase is still inducible. We have isolated mutants, containing alterations in a novel E. coli gene, ampG, in which a cloned C. freundii ampC gene is unable to respond to beta-lactam inducers. The ampG gene was cloned, sequenced and mapped to minute 9.6 on the E. coli chromosome. The deduced amino acid sequence predicted AmpG to be a 53 kDa, transmembrane protein, which we propose acts as a signal transducer or permease in the beta-lactamase induction system. Immediately upstream of ampG there is another 579-base-pair-long open reading frame (ORF) encoding a putative lipoprotein shown to be non-essential for beta-lactamase induction. We have found that ampG and this ORF form an operon, whose promoter is located in front of the ORF. Located closely upstream of the putative promoter is the morphogene bolA, which is transcribed in the opposite orientation. However, using transcription fusions, we have found that the ampG transcription is not regulated by bolA. In addition, we show that transcription is probably not regulated by either the starvation specific sigma factor RpoS, which controls bolA, or by AmpD the negative regulator for ampC transcription. PMID- 8231805 TI - Formation of several bacterial c-type cytochromes requires a novel membrane anchored protein that faces the periplasm. AB - We report here the discovery of a novel bacterial gene (cycH) whose product is involved in the biogenesis of most of the cellular cytochromes c. The cycH gene was detected in the course of characterizing a cytochrome oxidase-deficient Bradyrhizobium japonicum Tn5 mutant (strain COX3) in which the transposon insertion disrupted cycH. All of the c-type cytochromes detectable in aerobically grown B. japonicum wild-type cells were absent in the COX3 mutant, with the exception of cytochrome c1. A secondary phenotypic effect was the spectroscopic absence of the aa3-type cytochrome c oxidase. The nucleotide sequence of the cloned wild-type cycH gene predicted a membrane-bound 369-amino-acid protein with an M(r) of 39727. Results from studies on its membrane topology suggested that approximately 110 N-terminal amino acids are involved in anchoring the protein in the membrane, whereas the remaining two-thirds of the protein are exposed to the periplasm. We postulate that the CycH protein plays an essential role in an as yet unidentified periplasmic step in the biogenesis of holocytochromes c, except that of cytochrome c1. PMID- 8231806 TI - Phosphorylation of Spo0A activates its stimulation of in vitro transcription from the Bacillus subtilis spoIIG operon. AB - The spoIIG operon of Bacillus subtilis codes for a sporulation-specific sigma factor, sigma E. In vivo expression of the spoIIG promoter is activated shortly after the onset of sporulation and is dependent on kinA, spo0F, spo0B and spo0A genes. The products of these genes have been shown to participate in a phosphorelay reaction in vitro, culminating in phosphorylation of the transcription factor, Spo0A. The effect of Spo0A phosphorylation on in vitro transcription from the spoIIG promoter was determined. Aliquots from phosphorelay reactions enhanced spoIIG promoter activity 10-fold in transcription assays and stimulation of transcription was dependent on Spo0A phosphorylation. Our results provide biochemical evidence that Spo0A and the phosphorelay form a signal transduction pathway which activates spoII gene expression in development. PMID- 8231807 TI - The transcriptional activator HlyU of Vibrio cholerae: nucleotide sequence and role in virulence gene expression. AB - HlyU upregulates expression of the haemolysin, HlyA, of Vibrio cholerae. DNA sequence analysis indicates that HlyU is an 11.9 kDa protein containing a putative helix-turn-helix motif and belonging to a family of small regulatory proteins, including NoIR (Rhizobium meliloti), SmtB (Synechococcus PCC 7942) and ArsR (plasmids R773, Escherichia coli; pI258, Staphylococcus aureus; and pSX267, Staphylococcus xylosus). An hlyU mutant was constructed by insertional inactivation, and found to be deficient in the production of both the haemolysin and a 28 kDa secreted protein. The mutant was assessed for virulence in the infant mouse cholera model, revealing a 100-fold increase in the LD50. This suggests that HlyU promotes expression of virulence determinant(s) in vivo. PMID- 8231808 TI - An unusually small gene required for sporulation by Bacillus subtilis. AB - We report the cloning and characterization of an unusually small gene called spoVM whose product is required for normal formation of the cortex and coat during sporulation in Bacillus subtilis. The spoVM gene is adjacent to, and in convergent orientation with, the B. subtilis homologue to the Escherichia coli gene for ribosomal protein L28. The spoVM open reading frame is only 26 codons in length and is capable of encoding a polypeptide of 3 kDa. The short length of spoVM was verified by means of complementation experiments with wild-type and deletion-mutated copies of the open reading frame and by engineering the synthesis of the spoVM gene product in E. coli. Transcription of spoVM was induced during the second hour of sporulation (approximately stage II) by the appearance of the sporulation RNA polymerase sigma factor, sigma E. Efficient transcription of spoVM additionally required the action of the sporulation DNA binding protein SpoIIID. Because spoVM was not strongly required for the transcription of several genes expressed at late times in development, its protein product is likely to play a morphogenetic rather than a regulatory role in sporulation. PMID- 8231809 TI - Intercellular signalling in Vibrio harveyi: sequence and function of genes regulating expression of luminescence. AB - Density-dependent expression of luminescence in Vibrio harveyi is regulated by the concentration of an extracellular signal molecule (autoinducer) in the culture medium. A recombinant clone that restored function to one class of spontaneous dim mutants was found to encode functions necessary for the synthesis of, and response to, a signal molecule. Sequence analysis of the region encoding these functions revealed three open reading frames, two (luxL and luxM) that are required for production of an autoinducer substance and a third (luxN) that is required for response to this signal substance. The LuxL and LuxM proteins are not similar in amino acid sequence to other proteins in the database, but the LuxN protein contains regions of sequence resembling both the histidine protein kinase and the response regulator domains of the family of two-component, signal transduction proteins. The phenotypes of mutants with luxL, luxM and luxN defects indicated that an additional signal-response system controlling density-dependent expression of luminescence remains to be identified. PMID- 8231810 TI - SyrD is required for syringomycin production by Pseudomonas syringae pathovar syringae and is related to a family of ATP-binding secretion proteins. AB - The syrD gene of Pseudomonas syringae pathovar syringae strain B301D-R was characterized and sequenced. The syrD open reading frame is 1695 bp long and encodes a predicted protein, SyrD, of approximately 63 kDa. Database searches revealed that SyrD shares a high degree of similarity with the ATP-binding cassette (ABC) superfamily of transporter proteins which are responsible for specific nutrient uptake and for secretion of certain cellular products in prokaryotes, and for multiple drug resistance in mammals. The amino acid sequence homology between SyrD and the ABC proteins was greatest at the conserved residues which constitute the ATP-binding cassette of these proteins; these residues lie in the hydrophilic C-terminal half of SyrD. The N-terminus of SyrD is predicted to be hydrophobic and to contain six membrane-spanning alpha-helices. syrD mutants of strain B301D-R were significantly less virulent than other syr mutants, were deficient in four large polypeptides thought to be components of a syringomycin synthetase complex, and showed reduced expression of a syrB-lacZ reporter gene fusion in trans. It is proposed that SyrD is a cytoplasmic membrane protein that functions as an ATP-driven efflux pump for the secretion of syringomycin. PMID- 8231811 TI - The virD4 gene is required for virulence while virD3 and orf5 are not required for virulence of Agrobacterium tumefaciens. AB - The virD operon of the resident Ti plasmid of Agrobacterium tumefaciens contains loci involved in T-DNA processing and undefined virulence functions. Nucleotide sequence of the entire virD operon of pTiC58 revealed similarities to the virD operon of the root-inducing plasmid pRiA4b and to that of the octopine-type plasmid pTiA6NC. However, comparative sequence data show that virD of pTiC58 is more akin to that of the pRiA4b than to that of the pTiA6NC. T7f10::virD gene fusions were used to generate polypeptides that confirm the presence of four open reading frames virD1, virD2, virD3, and virD4 within virD which have a coding capacity for proteins of 16.1, 49.5, 72.6, and 73.5 kDa, respectively. virD3 therefore encodes a polypeptide 3.4 times larger (72.6 versus 21.3 kDa) than that encoded by virD3 of octopine Ti plasmids. Non-polar virD4 mutants could not be complemented by a distant homologue, TraG protein of plasmid RP4. An independently regulated fifth ORF (orf5) is located immediately downstream of 3' end of virD4 and encodes a polypeptide of 97.4 kDa. The expression of orf5 is dependent on its own promoter and is independent of acetosyringone induction in A. tumefaciens. Recently, it has been shown that virD3 of octopine Ri or Ti plasmids is not required for virulence. In this report, we confirm and extend these findings on a nopaline Ti plasmid by using several virD non-polar mutants that were tested for virulence. virD3 and orf5 non-polar mutants showed no effect on tumorigenicity on 14 different plant species, while virD4 mutants lost their tumorigenicity completely on all these test plants. These data suggest that virD3 and orf5 are not essential for virulence whereas virD4 is absolutely required on a wide range of host plants. PMID- 8231812 TI - Suppression of a dominant G-protein beta-subunit mutation in yeast by G alpha protein expression. AB - SCG1/GPA1, STE4 and STE18 encode the alpha, beta and gamma components of the G protein involved in mating pheromone signal transduction in Saccharomyces cerevisiae. Responses, including G1 arrest and expression of genes such as FUS1, are activated by beta gamma, which is negatively controlled by alpha(GDP). We previously demonstrated that overexpression of Scg1 suppresses responses to alpha factor and that expression of certain hybrids between Scg1 and mammalian G alpha proteins has the same effect and also suppresses growth arrest in an scg1-null mutant. Effects were attributed to sequestration of beta gamma. We now show that effects on growth rate, morphology and FUS1 expression are consistent with this model. The STE4HPL allele causes dominant activation of the response pathway, and is presumed to encode a beta subunit insensitive to control by alpha(GDP). Scg1 overexpression suppresses the growth arrest due to STE4HPL; normal alpha-factor responses and fertility are restored. A model based on sequestration of beta gamma reconciles this result with the apparent paradox that the same level of Scg1 overexpression inhibits responses and mating in wild-type cells. A G alpha i hybrid also restores growth and allows inefficient mating in the STE4HPL strain. PMID- 8231813 TI - Isozymes of S-adenosylmethionine synthetase are encoded by tandemly duplicated genes in Escherichia coli. AB - The sole biosynthetic route to S-adenosylmethionine, the primary biological alkylating agent, is catalysed by S-adenosylmethionine synthetase (ATP:L methionine S-adenosyltransferase). In Escherichia coli and Salmonella typhimurium numerous studies have located a structural gene (metK) for this enzyme at 63 min on the chromosomal map. We have now identified a second structural gene for S adenosylmethionine synthetase in E. coli by DNA hybridization experiments with metK as the probe; we denote this gene as metX. The metX gene is located adjacent to metK with the gene order speA metK metX speC. The metK and metX genes are separated by approximately 0.8 kb. The metK and the metX genes are oriented convergently as indicated by DNA hybridization experiments using sequences from the 5' and 3' ends of metK. The metK gene product is detected immunochemically only in cells growing in minimal media, whereas the metX gene product is detected immunochemically in cells grown in rich media at all growth phases and in stationary phase in minimal media. Mutants in metK or metX were obtained by insertion of a kanamycin resistance element into the coding region of the cloned metK gene (metK::kan) followed by use of homologous recombination to disrupt the chromosomal metK or metX gene. The metK::kan mutant thus prepared does not grow on minimal media but does grow normally on rich media, while the corresponding metX::kan mutant does not grow on rich media although it grows normally on minimal media. These results indicate that metK expression is essential for growth of E. coli on minimal media and metX expression is essential for growth on rich media. Our results demonstrate that AdoMet synthetase has an essential cellular and/or metabolic function. Furthermore, the growth phenotypes, as well as immunochemical studies, demonstrate that the two genes that encode S adenosylmethionine synthetase isozymes are differentially regulated. The mutations in metK and metX are highly unstable and readily yield kanamycin resistant cells in which the chromosomal location of the kanamycin-resistance element has changed. PMID- 8231814 TI - An outer membrane protein (OmpA) of Escherichia coli can be translocated across the cytoplasmic membrane of Bacillus subtilis. AB - The translocation of secretory proteins derived from a Gram-positive (Staphylococcus hyicus prolipase) or a Gram-negative (Escherichia coli pre-OmpA protein) bacterium across the cytoplasmic membrane was studied in E. coli and Bacillus subtilis. In both microorganisms, the prolipase was found to be secreted across the plasma membrane when either the pre-prolipase signal peptide (38 amino acids in length) or the pre-OmpA signal peptide (21 amino acids in length) was used. Expression of the gene encoding the authentic pre-OmpA protein in B. subtilis resulted in the translocation of mature OmpA protein across the plasma membrane. Processing of the OmpA precursor in B. subtilis required the electrochemical potential and was sensitive to sodium azide, suggesting that the B. subtilis SecA homologue was involved in the translocation process. The mature OmpA protein, which was most likely present in an aggregated state, was fully accessible to proteases in protoplasted cells. Therefore, our results clearly demonstrate that an outer membrane protein can be secreted by B. subtilis, supporting the notion that the basic mechanism of protein translocation is highly conserved in Gram-positive and Gram-negative bacteria. PMID- 8231815 TI - Sequence and molecular analysis of the nifL gene of Azotobacter vinelandii. AB - In both Klebsiella pneumoniae and Azotobacter vinelandii the nifL gene, which encodes a negative regulator of nitrogen fixation, lies immediately upstream of nifA. We have sequenced the A. vinelandii nifL gene and found that it is more homologous in its C-terminal domain to the histidine protein kinases (HPKs) than is K. pneumoniae NifL. In particular A. vinelandii NifL contains a conserved histidine at a position shown to be phosphorylated in other systems. Both NifL proteins are homologous in their N-termini to a part of the Halobacterium halobium bat gene product; Bat is involved in regulation of bacterio-opsin, the expression of which is oxygen sensitive. The same region showed homology to the haem-binding N-terminal domain of the Rhizobium meliloti fixL gene product, an oxygen-sensing protein. Like K. pneumoniae NifL, A. vinelandii NifL is shown here to prevent expression of nif genes in the presence of NH+4 or oxygen. The sequences found homologous in the C-terminal regions of NifL, FixL and Bat might therefore be involved in oxygen binding or sensing. An in-frame deletion mutation in the nifL coding region resulted in loss of repression by NH+4 and the mutant excreted high amounts of ammonia during nitrogen fixation, thus confirming a phenotype reported earlier for an insertion mutation. In addition, nifLA are cotranscribed in A. vinelandii as in K. pneumoniae, but expression from the A. vinelandii promoter requires neither RpoN nor NtrC. PMID- 8231816 TI - Molecular characterization of a fungal secondary metabolism promoter: transcription of the Aspergillus nidulans isopenicillin N synthetase gene is modulated by upstream negative elements. AB - The Aspergillus nidulans IPNS gene, encoding isopenicillin N synthetase, is a secondary metabolism gene. It is contiguous to, but divergently transcribed from, the ACVS gene at the penicillin gene cluster. The untranslated region between both ORFs is 872bp long. Here we present the physical and functional characterization of the IPNS transcriptional unit. Transcriptional start point (tsp) mapping reveals heterogeneity at the 5'-end of the mRNA, with a major start at -106 relative to the initiation codon. This indicates that the actual length of the non-transcribed intergenic region is 525bp. Functional elements in the IPNS upstream region have been defined by assaying beta-galactosidase activity in extracts from recombinant strains carrying deletion derivatives of the IPNS promoter fused to lacZ, integrated in single copy at the argB locus. Strains were grown in penicillin production broth under carbon catabolite repressing or derepressing conditions. The results of deletion analysis indicate that: (i) the IPNS promoter is mostly regulated by negative controls that act upon a high basal activity; (ii) sequential deletion of three of the negative cis-acting elements results in a mutated promoter that is 40 times (sucrose broth) or 12 times (lactose broth) more active than the wild type; (iii) one of these negative cis acting elements is involved in sucrose repression. Strikingly, it is located outside the non-transcribed 525bp intergenic region and maps to the coding region of the divergently transcribed ACVS gene; (iv) a 5'-deletion up to -56 (relative to the major tsp) contains information to provide almost half of the maximal promoter activity and allows initiation of transcription at the correct site. By using total-protein extracts from mycelia grown under penicillin producing conditions we have detected a DNA-binding activity that specifically shifts a promoter fragment located between -654 and -455 (relative to IPNS tsp). Deletions covering this region partially abolish IPNS promoter activity. The fragment in question overlaps the ACVS tsp. PMID- 8231817 TI - The maturation pathway of microcin B17, a peptide inhibitor of DNA gyrase. AB - The maturation pathway of microcin B17 (MccB17), a ribosomally synthesized peptide antibiotic which inhibits DNA gyrase, has been characterized. Synthesis of MccB17 involves several steps beginning with the translation of the MccB17 structural gene, mcbA, to yield a 69 amino acid precursor, preMccB17. Pre-MccB17 is then modified and folded by the action of three gene products, McbBCD, to yield proMccB17. Mutations in mcbA were isolated that permit modifications of the resulting mutant peptides, but prevent folding, suggesting that modification and folding are sequential steps. ProMccB17 is subsequently converted to MccB17 by removal of the N-terminal 26-amino-acid leader by a chromosomally encoded protease. Removal of the leader resulted in aggregation of the peptide, suggesting that the leader may function to maintain peptide solubility during synthesis in the cell. Finally, polyclonal antibodies raised against MccB17 recognize both MccB17 and proMccB17, but do not recognize preMccB17. This demonstrates the dramatic structural changes that result from the modifications and has been used to distinguish intermediates in the steps of maturation. PMID- 8231818 TI - Comparative mutagenicity of structurally related aliphatic epoxides in a modified Salmonella/microsome assay. AB - Four structurally related aliphatic epoxides (1,2-epoxypropane, 1,2 epoxyisobutane, cis- and trans-2,3-epoxybutane) have been tested in the Salmonella/microsome assay, modified for volatile substances, using the strains TA1535 and TA100. The aim of the study was to evaluate the effect of methylation on the mutagenicity of 1,2-epoxypropane in this vaporization assay, with and without exogenous metabolization. All substances induced a significant increase of revertants in the strains TA1535 and TA100. In terms of mutagenic potency, the following hierarchy was observed in the standard tester strain TA1535 and in the absence of rat S9: 1,2-epoxy-propane >> cis-2,3-epoxybutane > 1,2-epoxyisobutane > trans-2,3- epoxybutane. After exogenous metabolization, the mutagenic response of 1,2-epoxyisobutane was substantially reduced, while a moderate decrease of cis 2,3-epoxybutane was observed in the presence of S9, as compared with the response without S9. No influence of the S9 on the mutagenic response of trans-2,3 epoxybutane was noticed in both strains TA1535 and TA100, while an increased response with 1,2-epoxypropane was observed in TA100 but not in TA1535. The results suggest that the vaporization assay may provide more relevant information concerning mutagenic potencies of gaseous or volatile compounds than the common treat-and-plate or preincubation assays. Moreover, it appears that mutagenicity theories, based only upon inductive effects of side groups, may not suffice to explain differences in mutagenicity. Sterical factors or differential interactions with metabolizing enzymes could also be important in the evaluation of mutagenic effects. PMID- 8231819 TI - Aneuploidy induction by water extract from Tripterygium hypoglaucum (Level) Hutch in mouse bone marrow cells. AB - The aneuploidy-inducing activity of a Chinese medicinal herb, Tripterygium hypoglaucum (level) Hutch (THH), was investigated by means of three cytogenetic end-points, i.e. C-mitotic (CM) effects, micronuclei (MN) and parallel chromosome structural aberration (CA) analyses in vivo. The CA analysis was expected to reflect the origins of MN induced by clastogens or aneugens. The experiments were performed on mouse bone marrow cells. The animals were treated with the crude water extracts of THH (single i.p. injection) in the dose range 120-686 mg/kg. Colchicine (COL) was taken as a positive control for its known aneuploidy inducing effects. THH showed similar genotoxic effects to COL in CM, MN and CA analyses: positive CM effects were observed accompanied with increases of mitotic index and frequencies of CM cells as well as decreased frequencies of anaphase in all of the THH-treated groups. The compound showed a positive MN response in bone marrow polychromatic erythrocytes but was negative in CA analyses. No sex differences were found in any treated group. The preliminary results suggested that THH is an aneuploidy inducer in mouse bone marrow cells under the present experimental conditions. PMID- 8231820 TI - Mutation induction by UV light in retroviral hprt cDNA integrated at various chromosomal positions in repair-deficient hamster cells. AB - Mutation induction by UV irradiation was studied in a retroviral vector integrated in one copy per cell at various chromosomal positions. As a mutational target, hamster hprt cDNA was present on the retroviral vector. To minimize the influence of repair we used repair-deficient hamster cells, V-H1 and UV5, as a recipient for the vector. There is no major influence of chromosomal position on UV-induced mutation frequency and spectrum because no statistically significant difference between mutation induction in retroviral cDNA copies integrated at different chromosomal sites was observed. However, a major difference was found in mutation induction between the endogenous hamster hprt gene and the retroviral cDNA copies. Most noticeable was the absence in the cDNA of the strong strand bias for mutation induction, which was reported for the endogenous hprt gene. Our results with the hprt cDNA exclude as a general phenomenon a difference in mutation induction for leading and lagging strand DNA replication, which was proposed as an explanation for this strand bias in the endogenous gene. The similarity of mutation induction in the different retroviral cDNA copies, all directly surrounded by the same DNA sequence elements, together with the marked difference between the mutation induction in the endogenous gene and the cDNA copies may point to an important role of chromatin structure in mutation induction. PMID- 8231821 TI - The micronucleus assay in mouse peripheral blood reticulocytes demonstrates the transmission of chromosomal instability induced by mitomycin C and benzo[a]pyrene. AB - The frequency of micronuclei (MN) in mouse peripheral blood reticulocytes supravitally stained with acridine orange was assessed at different time intervals after treatment in vivo with two dose levels of mitomycin C (MMC) and benzo[a]pyrene. Increased frequencies were observed for many days after treatment, indicating that MN may be produced as a consequence of chromosomal instability transmitted by proliferating erythroblasts. These results confirm previous evidence of the persistent cytogenetic effects of MMC and suggest that clastogenic agents acting by different primary lesions may have a similar ability to induce this effect. PMID- 8231822 TI - Mutagenic specificity of the base analog 6-N-hydroxylaminopurine in the URA3 gene of the yeast Saccharomyces cerevisiae. AB - The mutational specificity of the base analog 6-N-hydroxylaminopurine (HAP) was studied in the URA3 gene of the yeast Saccharomyces cerevisiae. Twenty-nine independent HAP-induced ura3 mutations were sequenced. GC-->AT transitions were found most frequently (21 out of 29) while AT-->GC transitions were less abundant (five out of 29). Three GC-->TA transversions were also detected. Two interesting features of DNA context were revealed for transition mutations. One third of the transitions occurred at one site within short direct imperfect repeats converting them to perfect repeats. A model involving complementary interaction of imperfect repeats is proposed to explain the origin of these mutations. Nearly all of the rest of the GC-->AT as well as the AT-->GC transitions were found in the runs of several identical base pairs, predominantly in the middle or at the 3' template nucleotide of (G)n and (A)n runs. PMID- 8231823 TI - Paracetamol inhibits UV-induced DNA repair in resting human mononuclear blood cells in vitro. AB - The effects of paracetamol on the repair of DNA damage in resting human peripheral mononuclear blood cells (MNC) in vitro were investigated by means of the alkaline elution technique. Low doses of UV light (254 nm, 3 J/m2) caused a transient increase in the amount of DNA single-strand breaks and alkali-labile sites (SSBs). Paracetamol (0.1-1.0 mM) present during post-irradiation incubation approximately doubled the maximum level of UV-induced (1-3 J/m2) SSBs and delayed the completion of repair. Although there were considerable variations between cells prepared from different donors, the level of UV-induced DNA SSBs was always higher with paracetamol. Hydroxyurea (0.3 mM), an inhibitor of ribonucleotide reductase, caused a similar increased accumulation and slow removal of SSBs, whereas cytosine-1-beta-D-arabinofuranoside (Ara C) (10 microM), an inhibitor of DNA polymerases, led to a steady accumulation of DNA SSBs. The increased levels of SSBs caused by paracetamol or hydroxyurea were both completely suppressed by concomitant addition of deoxyribonucleosides; this supports the notion that paracetamol as well as hydroxyurea inhibits ribonucleotide reductase. About the same rates of formation and removal of UV-induced SSBs were observed in T lymphocytes, B lymphocytes and monocytes. In both isolated T lymphocytes and B lymphocytes, paracetamol (0.3 mM) markedly increased the level of DNA SSBs induced by UV, whereas monocytes seemed to be less sensitive to the effect of paracetamol. It is concluded that the inhibition of DNA repair may contribute to the clastogenic effects of paracetamol. PMID- 8231824 TI - Mutagenic activity of the antitumour agent homo-aza-steroidal ester of p-N,N bis(2-chloroethyl)aminophenoxyacetic acid (NSC 294859) in the Salmonella/microsome assay. AB - The mutagenic activity of the new antitumour agent 3 beta-hydroxy-13 alpha-amino 13,17-seco-5 alpha-androstan-17-oic-13,17-lactam- p-N,N-bis(2 chloroethyl)aminophenoxyacetate (NSC 294859) was studied in the Salmonella/microsome assay. It was found to induce base pair substitutions, causing dose-dependent increases in his+ revertants in strains TA100 and TA1535. The alkylating moiety, p-N,N-bis(2-chloroethyl)-aminophenoxyacetic acid, was shown to be less effective than the parent compound, while the modified steroid moiety, 3 beta-hydroxy-13 alpha-amino-13,17-seco-5 alpha-androstan-17-oic-13,17 lactam, showed no mutagenic effect in all strains used. The presence of metabolic activation enzymes in the test system induced a further increase in his+ revertants in strains TA100 and TA1535, in both the parent compound and the alkylating moiety of the parent compound, while it had no effect in the case of the steroidal lactam. PMID- 8231825 TI - Flow cytometric analysis of bromodeoxyuridine-induced micronuclei. AB - The effects of DNA substitution by the thymidine analogue 5-bromodeoxyuridine (BrdU) on cell cycle progression and micronucleus induction were studied in different mammalian cell cultures. Simultaneous flow cytometric measurements of DNA content and side scatter of nuclei in Chinese hamster embryo (CHE) cells revealed a concentration-dependent temporary block in the G2/M phase of the first cell cycle. NIH 3T3 cells and human amniotic fluid fibroblast-like cells, on the contrary, did not show any cell cycle disturbances in the presence of BrdU. Micronucleus frequency increased as soon as CHE cells started to divide and reached a plateau when all cells have divided. The height of this plateau was almost equal for 60 and 100 microM BrdU. This saturation of micronucleus induction was due to a saturation of BrdU incorporation into DNA already at a doses of 60 microM as shown by the BrdU/Hoechst quenching technique. Indirect immunofluorescent staining of kinetochores with CREST antibodies revealed that nearly all BrdU-induced micronuclei were kinetochore-negative suggesting the presence of acentric chromosome fragments in these micronuclei. DNA distributions of micronuclei measured by flow cytometry showed several peaks representing micronuclei which contain DNA fragments of defined sizes induced by non-random breakage of chromosomes 1 and X as verified by flow karyotyping and C-banding. PMID- 8231826 TI - Persistence of sister chromatid exchange by 9-beta-D-arabinofuranosyladenine in Chinese hamster ovary cells cultivated in vitro. AB - The cytogenetic activity of 9-beta-D-arabinofuranosyladenine (ara-A), a known antitumor agent with an unknown mechanism of action (possibly acting via demethylation and/or decrease in DNA synthesis), has been tested in Chinese hamster ovary (CHO) cells cultivated in vitro, with respect to its ability to induce sister chromatid exchange (SCE). The agent shows no effect on cell cycle parameters (proliferation and mitotic indices) at concentrations up to 100 microM, when administered in pulses ranging from 3 to 12 h. Furthermore, even if administered over the entire treatment period (24 h), a severe cell cycle delay appears only at a concentration of 100 microM. A clear increase in the SCE frequency is produced starting from a concentration of 10 microM, irrespective of the treatment protocol (i.e. pulse versus continuous). This effect on the SCE frequency is maintained for as long as 10 cell cycles after removal of the agent. A partial inactivation of S-adenosyl-L-homocysteine (AdoHcy) hydrolase seems to be produced for as long as 6 h after agent removal. We suggest that under our experimental conditions ara-A exerts its action through a heritable epigenetic modification. PMID- 8231827 TI - An investigation of micronucleus and mutation induction by oxazepam in mammalian cells. AB - The benzodiazepines are a class of drugs that are widely used in the treatment of various psychiatric disorders. One member of this class, oxazepam, is also a common metabolite of several other benzodiazepines. Since the evidence for the genetic toxicity and carcinogenic properties of these compounds is inconsistent, we investigated the oxazepam-induced formation of micronuclei in Syrian Hamster embryo fibroblast (SHE) cells, human amniotic fluid fibroblast-like (AFFL) cells and L5178Y mouse cells. A dose-dependent increase in micronucleus fractions was found in all three cell lines. The time course of micronucleus induction in L5178Y cells showed a maximum at 5 h after treatment, suggesting that the micronuclei were formed in the first mitosis after treatment. Kinetochore staining (CREST-antiserum) revealed the presence of kinetochores in approximately 50% of the micronuclei in all three cell types. This result was further confirmed by in situ hybridization in L5178Y cells and indicates the presence of whole chromosomes or centric fragments as well as acentric fragments in the oxazepam induced micronuclei. The L5178Y cells did not show a mutagenic response to oxazepam at any of the doses or expression times used. PMID- 8231828 TI - Elevated intracellular dCTP levels reduce the induction of GC-->AT transitions in yeast by ethyl methanesulfonate or N-methyl-N'-nitro-N- nitrosoguanidine but increase alkylation-induced GC-->CG transversions. AB - The effect of an increased intracellular dCTP:dTTP ratio on the specificities of ethyl methanesulfonate (EMS) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) mutagenesis was examined in the yeast Saccharomyces cerevisiae. To do so, we used a dCMP deaminase-deficient (dcd1) strain having a dCTP:dTTP ratio > 77-fold larger than its isogenic wild-type parent under the treatment conditions employed. This DNA precursor imbalance lowered the frequencies of EMS- or MNNG induced SUP4-o mutations by 75 or 45%, respectively, relative to the corresponding values for the wild-type strain. A total of 405 SUP4-o mutations produced by the alkylating agents in the dcd1 background were characterized by DNA sequencing and the mutational spectra were compared to those for 399 mutations induced in the wild-type parent and 207 mutations that arose spontaneously in the dcd1 strain. Unexpectedly, the frequencies of EMS- and MNNG induced GC-->AT transitions in the dcd1 strain were found to be reduced by 93 and 68%, respectively, considerably more than the decreases for the overall SUP4-o mutation frequencies. The differences were due mainly to substantial increases in the frequencies of GC-->CG transversions. Although these events were the predominant type of spontaneous substitution in the dcd1 strain, they were more frequent after alkylation treatment and were distributed differently than the spontaneous GC-->CG transversions. Preferences for the EMS- or MNNG-induced GC- >AT transitions to occur at GC sites having the guanine located on the transcribed strand or preceded by a 5' purine, respectively, also were diminished in the dcd1 strain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231829 TI - Evidence for preferential repair of 3-carbethoxypsoralen plus UVA induced DNA lesions in the active MAT alpha locus in Saccharomyces cerevisiae using the UvrABC assay. AB - The occurrence of preferential repair in Saccharomyces cerevisiae of the active MAT alpha locus compared with the inactive HML alpha locus was confirmed after 254 nm UV irradiation. Experiments carried out using the UvrABC excinuclease assay with the monofunctional furocoumarin 3-carbethoxypsoralen (3-CPs) plus UVA radiation which induce mainly monoadducts in DNA demonstrated preferential repair of the active MAT alpha locus compared with the inactive HML alpha locus in a SIR+ strain. However, as after 254 nm UV irradiation, no difference in the rate of removal of 3-CPs plus UVA induced lesions was observed between the two loci in the sir-3 mutant in which both loci are active. Thus, it appears that 3-CPs plus UVA induced monoadducts as well as pyrimidine dimers are subject to preferential repair. PMID- 8231830 TI - Insulin-like growth factor-I receptor increases in aortic endothelial cells from diabetic rats. AB - Endothelial cells are likely to play an important role in the development of diabetic vascular diseases, since they are exposed directly to the abnormal circulating metabolites of diabetes and may be easily damaged early in the natural course of vascular complications. In this study, aortic endothelial cells were cultured from diabetic BB rats. Their binding and internalization of insulin like growth factor-I (IGF-I) were measured. IGF-I binding was higher in cells of diabetic rats than of control rats at both 37 degrees C (4.5% +/- 1.6% v 2.74% +/ 0.9% per mg protein, P < .05) and 4 degrees C (20.6% +/- 5.6% v 13.7% +/- 4.6% per mg protein, P < .01). Internalization of IGF-I also increased (1.62% +/- 0.2% v 0.74% +/- 0.15% of total count at 37 degrees C after 60 minutes, P < .05). Cross-linking studies showed that in cells from diabetic rats, the major band of 140 kd corresponding to the alpha-subunit of the IGF-I receptor increased in density by 50% compared with those from control rats. The IGF-I-stimulated tyrosine kinase activity (TKA) of partially purified receptor from cells of diabetic rats, measured using poly-glu-tyr as substrate, was normal. Since the biological effects of IGF-I are initiated by its binding to the IGF-I receptor, which is able to transduce mitogenic and metabolic signals, our results support the hypothesis that the IGF-I receptor is involved in the development of diabetic vascular complications. PMID- 8231831 TI - Metabolism of cholesterol and apolipoprotein B in celiac disease. AB - To test the association of cholesterol malabsorption with cholesterol and lipoprotein metabolism, we determined low-density lipoprotein (LDL) apolipoprotein (apo) B kinetics simultaneously with measurements of cholesterol absorption and synthesis in six patients with celiac disease (CD) before and during the gluten-free diet (GFD). The basal condition was characterized by low cholesterol absorption, enhanced cholesterol synthesis, and high removal and transport rate of LDL apo B. The GFD markedly improved cholesterol absorption and decreased intestinal influx of cholesterol, fecal neutral steroids, and cholesterol synthesis. Of plasma total and lipoprotein cholesterol levels, only plasma high-density lipoprotein (HDL) was enhanced by the GFD proportionately to cholesterol absorption. The plasma LDL apo B level remained unchanged because of simultaneous decreases in the fractional catabolic rate (FCR) and transport rate of LDL apo B. In fact, the more cholesterol absorption was improved by the GFD, the more the FCR and transport rate for LDL apo B were decreased, and their reductions were closely related to the decrease in cholesterol synthesis. The present results show that cholesterol absorption, cholesterol synthesis, hepatic B/E receptor activity, and LDL apo B transport rate are closely associated with each other and that their levels can change markedly with no detectable change in serum levels of LDL cholesterol or apo B. PMID- 8231832 TI - The effects of biotin deficiency on organic acid metabolism: increase in propionyl coenzyme A-related organic acids in biotin-deficient rats. AB - Volatile organic acid levels in plasma and tissues and nonvolatile organic acid levels in urine of biotin-deficient (BD) rats were measured and compared with other factors of biotin deficiency. Biotin levels and the activities of propionyl coenzyme A (CoA) carboxylase (PCC) in the livers of these rats were decreased, respectively, to 22% +/- 3% and 3.6% +/- 0.3% of the average values of pair-fed controls. Plasma concentrations of propionate were higher (15 to 223 micrograms/mL) than those of controls (5 to 7 micrograms/mL), whereas plasma levels of 3-methylcrotonate were only minimally increased as compared with those of controls. Concentrations of these volatile acids in the tissues were similarly increased, although those in brain showed less remarkable increases as compared with levels in other tissues. In the urine of BD rats, large amounts of organic acids derived from propionyl CoA, as well as those from 3-methylcrotonyl CoA, were excreted. Plasma propionate levels were not apparently related to the severity of clinical symptoms, biotin levels, or carboxylase activities, but were related to the amounts of urinary ketone bodies, lactate, and some of the organic acids derived from branched-chain amino acids, including those from propionyl CoA. PMID- 8231833 TI - Glucose modulation of islet monoamine oxidase activity in lean and obese hyperglycemic mice. AB - Islet beta-cell monoamines are known to influence the insulin-releasing mechanisms. These amines are localized in the insulin-secretory granules and are inactivated by the enzyme monoamine oxidase (MAO), a hydrogen peroxide (H2O2) generating enzyme. The activity of islet MAO may consequently be of importance for insulin secretion. In the present investigation, we studied the relation between islet MAO activity and plasma levels of insulin and glucose in obese (ob/ob) hyperglycemic mice and their lean littermates. In addition, the effect of glucose on the MAO activity of in vitro-cultured islets was studied. MAO activity was assayed with serotonin, dopamine (DA), and beta-phenylethylamine (PEA) as substrates. After an overnight fast in adult (age, 6 months) lean mice, islet MAO activity was increased by 35% to 70%. Plasma levels of glucose and insulin were markedly decreased as expected. However, fasting in adult obese mice either did not affect islet MAO activity (PEA and DA) or induced a slight decrease (serotonin) of approximately 25% (P < .05). Plasma glucose levels in adult obese mice were not significantly affected by the overnight fast. However, a correlation analysis based on individual adult obese mice (fed and fasted) showed a negative correlation between plasma glucose concentration and islet MAO activity with PEA (r = -.65, P < .02) and DA (r = -.66, P < .02), respectively. Further, a positive correlation (r = +.58, P < .05) was found between glucose level and islet MAO activity when using serotonin as substrate. There was no difference in islet MAO activity with PEA and DA as substrates in fed obese versus fed lean mice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231834 TI - Regional fat distribution and metabolism in a new mouse model (C57BL/6J) of non insulin-dependent diabetes mellitus. AB - It has been suggested that a genetic predisposition and an increased total fat mass, particularly a specific increase in visceral fat, contribute to the metabolic aberrations associated with human non-insulin-dependent diabetes mellitus (NIDDM). In this study, we investigated the interactions between genetic and dietary components on fat distribution and metabolism in two mouse strains, one genetically predisposed to NIDDM (BL/6) and one not (A/J), fed either a chow diet or a high-fat, high-simple carbohydrate (HFHSC) diet for 5 months. As expected, both strains of mice fed a HFHSC diet were heavier, had more fat in both the subcutaneous (inguinal [ING] and visceral (mesenteric [MES]) regions, and had larger fat cells and higher lipoprotein lipase (LPL) activities. The results of interactions between strain and diet showed important differences in fat distribution and metabolism between strains. In comparison with A/J mice, BL/6 mice fed a HFHSC diet developed hyperglycemia, hyperinsulinemia, and hypercholesterolemia, were heavier, had more overall fat, and particularly increased their MES adipose tissue. This increase in visceral fat mass was due to an increase in fat cell number. In contrast, BL/6 mice fed a chow diet had less overall fat, a smaller MES fat pad with smaller adipocytes, and lower LPL activity than A/J controls. Significant differences between BL/6 and A/J mice fed either a HFHSC or a chow diet were not observed in ING adipose tissue. These data suggest that in BL/6 mice, changes in the metabolic characteristics of visceral fat seem to be a specific characteristic associated with the genetic predisposition for NIDDM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231835 TI - Inheritance conjointly contributing to fibrinolysis and hyperlipidemia. AB - Our specific aim was to assess within-family relationships of basal fibrinolytic activity and its determinants in hyperlipidemic probands (n = 34) with high lipoprotein (a) [Lp(a)] levels (> 35 mg/dL) and their first-degree relatives (n = 74) and in hyperlipidemic probands (n = 19) with Lp(a) < 35 and their first degree relatives (n = 23). Probands' plasminogen activator inhibitor activity (PAI-Fx), the major fibrinolysis inhibitor, correlated with first-degree relatives' PAI-Fx in high-Lp(a) kindreds (r = .30, P = .06) and in Lp(a) < 35 kindreds (r = .43, P < or = .05). Probands' tissue plasminogen activator activity (tPA-Fx), the major fibrinolysis activator, was inversely associated with first degree relatives' PAI-Fx in high-Lp(a) kindreds (r = -.30, P = .06) and in Lp(a) < 35 kindreds (r = -.49, P < or = .025). These correlations [irrespective of probands' Lp(a)] pointed to within-family heritability of the major fibrinolysis inhibitor, PAI-Fx, and the fibrinolysis stimulator, tPA-Fx. There were many other within-family correlations. High-Lp(a) probands' tPA-Fx, the stimulator of fibrinolysis, correlated with first-degree relatives' tPA-Fx (r = .32, P < or = .05). High-Lp(a) probands' plasminogen was inversely correlated with first-degree relatives' alpha 2-antiplasmin, a major fibrinolytic inhibitor (r = -.41, P < or = .01), and with their Lp(a) [r = -.24, P < or = .05]. High-Lp(a) probands' tPA Fx correlated inversely with first-degree relatives' apolipoprotein (apo) B (r = .28) and triglyceride ([TG] r = -.41), and positively with their high-density lipoprotein cholesterol ([HDLC] r = .40) and apo A-1 (r = .33; all P < or = .025). High-Lp(a) probands' PAI-Fx correlated positively with first-degree relatives' apo B (r = .34) and TG (r = .47), and inversely with HDLC (r = -.34) and apo A-1 (r = -.30; all P < or = .01). By stepwise regression, the Quetelet index (a measure of relative ponderosity) was independently inversely associated with tPA-Fx (P < or = .05) and positively associated with tPA-Ag and PAI-Fx (P < or = .05). TG was a positive independent determinant of PAI-Fx (P < or = .05), alpha 2-antiplasmin (P < or = .05), and plasminogen (P < or = .05). Lp(a) was a positive, independent determinant of fibrinogen (P < or = .05).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8231836 TI - Quantitation of the pathways followed in the conversion of fructose to glucose in liver. AB - Recently, only about 50% of the conversion of fructose to glucose was reported to be via fructose-1-P aldolase catalysis in children. This was also suggested to be the case in adults. That possibility has been tested using a method that quantifies the pathways of fructose conversion to glucose via the fate of 14C from specifically labeled fructose. Trace [6-14C] fructose or its immediate precursor [6-14C]sorbitol with unlabeled fructose (0.3 mg/kg body weight/min) was given intravenously or intragastrically with trace [1-14C]lactate to six normal adults fasted overnight. The distributions of 14C in glucose from blood samples were determined. The ratios of 14C in C1 to C6 of the glucose were equal to or only slightly less than the ratios of 14C in C3 to C4. Since incorporation into C3 and C4 of glucose must have arisen via the conversion of [1-14C]lactate to [1 14C]triose phosphates, fructose conversion to glucose must also have arisen predominantly via the triose phosphates. From the ratios, 85.1% to 100%, a mean of 94.9% of the fructose converted to glucose is calculated to have been converted to glucose with cleavage of the carbon skeleton of the fructose. These findings contrast with the report that in children under similar conditions only about 50% of the conversion of fructose to glucose is with cleavage. The findings agree with previous results in which fructose was administered to normal adults as a bolus at a dose of 60 mg/kg body weight. The possible reasons that the findings in children are different from those in adults are considered. PMID- 8231837 TI - Selective impairment of the cytoplasmic Ca2+ response to glucose in pancreatic beta cells of streptozocin-induced non-insulin-dependent diabetic rats. AB - Pancreatic islets from the streptozocin-induced non-insulin-dependent diabetes mellitus (NIDDM) rat model showed a diminished insulin response to 16.7 mmol/L glucose, but the insulin response to arginine remained intact. To evaluate the importance of intracellular calcium concentration ([Ca2+]i) in the diminished insulin response to glucose, the [Ca2+]i of pancreatic beta cells was investigated using fura-2. Glucose produced heterogeneous responses of [Ca2+]i, which were in beta-cell clusters of both the control and NIDDM groups. Many cells showed initial slight decreases of [Ca2+]i, which were followed by gradual and large increments of [Ca2+]i after glucose stimulation of beta cells in the control group. On the other hand, the increase of [Ca2+]i in response to glucose was markedly diminished in beta cells of the NIDDM group compared with controls. The average lag time to [Ca2+]i elevation of beta cells in the NIDDM group was significantly longer than that of the control group. Arginine produced marked increases of [Ca2+]i, in contrast to the effect of glucose stimulation in the NIDDM group. These results suggest that the diminished and delayed [Ca2+]i increases in beta cells of NIDDM rats in response to glucose stimulation are responsible for the selectively impaired insulin response to glucose in the rat model of NIDDM. PMID- 8231838 TI - Increased production of apolipoprotein A-I associated with elevated plasma levels of high-density lipoproteins, apolipoprotein A-I, and lipoprotein A-I in a patient with familial hyperalphalipoproteinemia. AB - Familial hyperalphalipoproteinemia (FHA) is a heritable trait associated with elevated plasma concentrations of high-density lipoprotein (HDL) cholesterol and possibly with longevity and protection against coronary heart disease (CHD). The metabolic basis and molecular etiology of FHA have not been established in most kindreds. The proband of a kindred with FHA and possible longevity was found to have elevated plasma levels of HDL cholesterol, apolipoprotein (apo) A-I, and lipoproteins containing apo A-I without apo A-II (Lp A-I), but normal levels of apo A-II and lipoproteins containing apo A-I with apo A-II (Lp A-I:A-II). The in vivo kinetics of apo A-I and apo A-II were studied in the FHA proband and in control subjects using both exogenous radiotracer (125I-apo A-I and 131I-apo A II) and endogenous stable isotope (primed constant infusion of 13C6 phenylalanine) labeling techniques. The production rate (PR) of apo A-I was markedly increased in the FHA subject (28.9 mg/kg.d) compared with the control subjects (12.0 +/- 2.1 mg/kg.d), whereas the apo A-II PR was not substantially increased. The primary sequence of the proband's apo A-I gene, including 1.2 kb of the 5'-flanking sequence, was normal. We conclude that a selective upregulation of apo A-I production is one metabolic cause of FHA, and results in high plasma concentrations of HDL cholesterol, apo A-I, and Lp A-I and possibly in protection from atherosclerotic CHD. PMID- 8231839 TI - Effect of diabetes on levels of lipid peroxides and glycolipids in rat brain. AB - The effects of diabetes on levels of lipid peroxides and glycolipids in brain were studied in alloxan (18 mg/100 g body weight) diabetic rats. Free fatty acid (FFA) and malondialdehyde (MDA) levels were increased in the brains of diabetic animals. On the other hand, activities of the antioxidative enzymes catalase and superoxide dismutase (SOD) were decreased. The study also showed elevated levels of most of the glycolipid fractions except gangliosides, which were found to decrease in diabetic brain. Administration of insulin to diabetic animals results in the restoration of these parameters to normal levels. These changes observed in diabetic brain may be responsible for the increased frequency of stroke in diabetes. PMID- 8231840 TI - Effects of doxazosin, a selective alpha 1-inhibitor, on plasma insulin and blood glucose response to a glucose tolerance test in essential hypertension. AB - Essential hypertension is characterized by an insulin-resistance state that may be responsible for the increased atherogenesis typical of this disease. To investigate the effect on glucose and insulin metabolism of doxazosin, an antihypertensive drug whose favorable impact on lipid metabolism is well known, 10 newly diagnosed essential hypertensive patients of the same age and body mass index (BMI) were selected for our study. They underwent an oral glucose tolerance test (OGTT) with measurement of plasma insulin levels before and after a 6-month treatment period with doxazosin. In this period of time, they took no other drugs and followed no diets, nor did their weights vary. The daily dose of doxazosin was increased (maximum, 8 mg) to achieve a normotensive blood pressure (< 140/90). At the end of treatment, total and low-density lipoprotein (LDL) cholesterol levels (225 +/- 18 v 200 +/- 16 and 177 +/- 8 v 150 +/- 7 mg/dL, respectively; P < .05) were decreased, confirming the lipid-lowering effect of the drug. The OGTT showed a significant decrease of plasma insulin (16.04 +/- 1.8 v 10.99 +/- 0.9 mU/mL.min, P < .05) and blood glucose (22.54 +/- 1.6 v 20.83 +/- 1.6 g/dL.min, P < .05) areas. The glucose to insulin ratio, also known as the insulin sensitivity index, increased (1.56 +/- 0.15 v 1.95 +/- 0.12, P < .05). These findings seem to provide evidence of a favorable effect of doxazosin on insulin action. PMID- 8231841 TI - Effects of treatment with recombinant human growth hormone on insulin sensitivity and glucose metabolism in adults with growth hormone deficiency. AB - In a double-blind, cross-over, placebo-controlled trial, the effect of 26 weeks of replacement therapy with recombinant human growth hormone (rhGH) on insulin sensitivity and glucose metabolism in nine patients with adult-onset growth hormone deficiency was studied with a euglycemic clamp. Glucose production and utilization were studied with D-(3-3H)-glucose infusions. Comparisons were made with placebo treatment for 6 and 26 weeks, respectively. GH therapy for 6 weeks increased fasting plasma concentrations of glucose and insulin. However, after 26 weeks of GH treatment, no significant changes in glucose or insulin concentrations were recorded. GH treatment induced a marked change in insulin action evident after 6 weeks of therapy as shown by lower glucose infusion rates (GIRs) during the clamp compared with placebo treatment (2.6 +/- 0.4 v 4.1 +/- 0.7 mg.kg-1.min-1). This change in insulin action was due to a decreased insulin effect on glucose utilization. After 26 weeks of GH therapy, there was no significant difference in GIRs. During placebo treatment, insulin sensitivity and insulin, glucose, and nonesterified fatty acid (NEFA) concentrations were unchanged compared with concentrations measured before the study. Thus GH replacement therapy induces a change in insulin action in GH-deficient individuals. Whether this change represents a decrease in insulin action (ie, insulin resistance) or a restoration of action to normal is presently unclear, since a healthy control group was not included in the study. During long-term treatment, the present study suggests that the change in insulin action can be reversed, probably secondarily to changes in body composition. PMID- 8231842 TI - Independent associations between plasma lipoprotein subfraction levels and the course of coronary artery disease in the St. Thomas' Atherosclerosis Regression Study (STARS). AB - Associations between plasma lipoprotein subfractions and changes in coronary artery diseases (CAD) were examined in 74 men who completed the St. Thomas' Atherosclerosis Regression Study (STARS). Plasma lipoproteins were isolated by stepwise, preparative ultracentrifugation at repeated intervals during the 38 month trial. Paired coronary angiograms were quantitatively analyzed by a computerized method. In univariate linear regression analysis, changes in mean absolute width (delta MAWS) and minimum absolute with (delta MinAWS) of coronary segments were significantly correlated with in-trial concentrations of cholesterol in intermediate-density lipoprotein ([IDL] d = 1.006 to 1.019 kg/L), low-density lipoprotein ([LDL2] d = 1.019 to 1.040 kg/L; LDL3, d = 1.040 to 1.063 kg/L), and high-density lipoprotein ([HDL3] d = 1.125 to 1.210 kg/L) subfractions; no significant associations were found with other lipoproteins. IDL, LDL3, and HDL3 cholesterol were then selected for multiple linear regression analysis because these variables were not co-correlated and because they attained a significance of P less than or equal to .1 in univariate regression. In this analysis, only LDL3 cholesterol level was a significant negative predictor (P < .05) of both delta MAWS and delta MinAWS; a positive association between delta MinAWS and HDL3 cholesterol level just failed to reach conventional statistical significance (P = .066). Correlations between changes in coronary luminal dimensions and LDL3 cholesterol level were independent of age, smoking, weight, and blood pressure. Most patients showing regression of coronary atherosclerosis had an LDL3 cholesterol level of less than 1.8 mmol/L. The findings suggest that LDL3 is the plasma lipoprotein subfraction that exerts the single most powerful effect on the course of CAD in middle-aged men with hypercholesterolemia. PMID- 8231843 TI - A naturally occurring furan fatty acid enhances drug inhibition of thyroxine binding in serum. AB - We studied the thyroxine (T4)-displacing effects of a naturally occurring, highly albumin-bound furanoid acid that accumulates in serum in renal failure to concentrations in excess of 0.2 mmol/L. This substance, 3-carboxy-4-methyl-5 propyl-2-furanpropanoic acid (CMPF), has been shown to displace acidic drugs from albumin binding. The effects of CMPF on ligand binding were assessed in the following systems: (1) T4 binding to T4-binding globulin (TBG) and transthyretin (TTR), (2) T4 binding in undiluted serum, (3) T4-displacing potency of fenclofenac, furosemide, diflunisal, and aspirin in undiluted serum, (4) serum binding of [14C]-drug preparations, and (5) serum binding of [14C]-oleic acid. CMPF had a minor direct effect on T4 binding to TBG comparable in relative affinity to that of aspirin, ie, almost 7 orders of magnitude less than T4 itself. CMPF alone at a concentration of 0.3 mmol/L, which produced only a 10% to 14% increase in free T4 augmented the T4-displacing effects of high therapeutic concentrations of the various drugs in undiluted serum as follows: furosemide by 180%, fenclofenac by 160%, diflunisal by 130%, and aspirin by 40%. In the presence of fenclofenac, increments of CMPF from 0.075 to 0.3 mmol/L progressively augmented the T4-displacing effect of this drug, associated with a progressive increase in its calculated free concentration. CMPF also inhibited the binding of [14C]-oleic acid, suggesting that in some situations CMPF could also indirectly influence thyroid hormone binding by increasing the unbound concentration of nonesterified fatty acids (NEFA), as previously described.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231844 TI - Reciprocal effects of the protein kinase C inhibitors staurosporine and H-7 on the regulation of glycogen synthase and phosphorylase in the primary culture of hepatocytes. AB - The effects of the protein kinase C inhibitors staurosporine and H-7 [1-(5 isoquinolinylsulfonyl)-2-methylpiperazine] on glucose-induced regulation of glycogen synthase and phosphorylase activities were investigated in the primary culture of hepatocytes. Glycogen synthesis as measured by the incorporation of [14C]glucose into glycogen was enhanced up to 78% (P < .001) by 100 nmol/L staurosporine. In contrast, H-7 inhibited glycogen synthesis in a dose-dependent manner, with an IC50 value of 70 mumol/L. Activation of glycogen synthase by 30 mmol/L glucose was enhanced significantly (P < .02 and less) by staurosporine at 20 nmol/L and higher concentrations whereas the activity of this enzyme was inhibited by H-7 (IC50 = 50 mumol/L). The inactivation of phosphorylase by glucose was significantly greater when staurosporine was included in the medium. However, H-7 increased the phosphorylase activity ratio by 1.5- to 2.5-fold at concentrations of 20 to 100 mumol/L. The time course of synthase activation and phosphorylase inactivation showed that the effect of glucose was enhanced by staurosporine and inhibited by H-7. These novel reciprocal effects of protein kinase C inhibitors were also observed at different concentrations of glucose. The effects of H-8, a compound with structural resemblance to H-7 and an inhibitor of protein kinase A, were similar to those of staurosporine but not to those of H-7. Staurosporine blocked the effects of vasopressin and 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA), whereas H-7 in combination with these protein kinase C activators acted in the same direction. The effects of staurosporine, a relatively more specific inhibitor of protein kinase C, indicated that this enzyme plays a role in the regulation of glycogen metabolism in liver. However, H-7, which is known to have protein kinase C-independent effects in intact cells, seems to alter the activities of glycogen synthase and phosphorylase by a different mechanism. PMID- 8231845 TI - Redox status and protein binding of plasma homocysteine and other aminothiols in patients with homocystinuria. AB - Elevations of homocyst(e)ine levels in the blood of patients with homocystinuria may explain the high cardiovascular morbidity. We determined levels of reduced, oxidized, and protein-bound homocyst(e)ine, cyst(e)ine, and cyst(e)inylglycine in plasma from eight patients with homocystinuria. The technique used involved trapping of reduced thiols by collecting blood directly into tubes containing sulfhydryl-reactive reagents. All patients had high levels of homocysteine (range, 1.9 to 91.2 mumol/L), and among the aminothiols investigated, this species showed the most drastic elevation compared with trace levels (< 0.4 mumol/L) found in healthy subjects. The ratio between free homocysteine and total homocyst(e)ine (reduced to total ratio) was above normal and positively correlated to the reduced to total ratio for cyst(e)ine, suggesting that an equilibrium exists between these species through sulfhydryl disulfide exchange. The other homocyst(e)ine species (oxidized and protein-bound) were also markedly increased in patients with homocystinuria. Plasma cysteine and cysteinylglycine levels were moderately increased, whereas plasma concentrations of protein-bound cyst(e)ine, protein-bound cyst(e)inylglycine, and free cystine were below normal. Homocysteine in particular and other homocyst(e)ine species are markedly increased in plasma of homocystinurics, and these changes are associated with pronounced alterations in the level and the redox status of other aminothiols. This should be taken into account when considering homocyst(e)ine as an atherogenic agent, and the role of various homocyst(e)ine species in the pathogenesis of homocystinuria. PMID- 8231846 TI - Effect of gemfibrozil on adipose tissue and muscle lipoprotein lipase. AB - To better understand the mechanism of action of gemfibrozil on plasma triglycerides, lipoprotein lipase (LPL) concentration was measured in adipose tissue and muscle of 16 hypertriglyceridemic patients before and after treatment with gemfibrozil for 6 weeks. The patients were divided into three groups based on clinical criteria as follows: group 1, hypertriglyceridemia without secondary factors; group 2, hypertriglyceridemia with diabetes; and group 3, hypertriglyceridemia with renal insufficiency. LPL activity, immunoreactive mass, synthetic rate, and mRNA levels were measured in the adipose tissue samples, and LPL activity and mass in the muscle samples. Serum triglyceride levels were decreased by 46% by gemfibrozil, and patients demonstrated no change in diet, weight, or glycohemoglobin during the 6 weeks of treatment. Despite the decrease of blood triglyceride levels, there was no significant change in any measure of LPL either in adipose tissue or muscle. Although several patients demonstrated increases in muscle LPL activity, these changes were inconsistent and not statistically significant. Because there was no significant change in LPL, we conclude that gemfibrozil in these patients decreased circulating triglyceride levels predominantly by decreasing hepatic very-low-density lipoprotein (VLDL) secretion. PMID- 8231847 TI - A concentration gradient of glucose from liver to plasma. AB - Concentrations of glucose in plasma water and liver water were determined in rats under a number of conditions. In fasted and fed postadsorptive rats, the concentration of glucose in plasma water averaged 5.5 +/- 0.5 and 6.8 +/- 0.2 mmol/L, respectively. The concentration in liver water was 8.2 +/- 0.9 and 11.1 +/- 1.1 mmol/L, respectively (mean +/- SD). The concentration ratio of liver water to plasma water (LW/PW) was 1.52 +/- 0.1 (range, 1.4 to 1.7; n = 7) in fasted rats and 1.64 +/- 0.39 (range, 1.38 to 2.0; n = 10 [mean +/- SD]) in fed rats. Fasted rats treated with glucagon contained no liver glycogen. The concentration of glucose in plasma water was 7.1 +/- 0.3 mmol/L and in liver water 9.2 +/- 0.5 mmol/L, and the LW/PW averaged 1.35 +/- 0.08 (range, 1.23 to 1.45; n = 8). Insulin-injected hypoglycemic rats contained very little glycogen. The LW/PW was 2.7 +/- 0.2. In fasted rats infused intragastrically with 30 mg/min/kg glucose, the concentration of glucose in mixed portal-arterial plasma water entering the liver averaged 10.3 +/- 0.8 mmol/L and that in liver water 12.4 +/- 1.3 mmol/L (mean +/- SD), with the LW/PW averaging 1.21 +/- 0.1 (range, 1.08 to 1.4; n = 8). Fasted rats were infused with 3.4 mg/min/kg sodium lactate labeled with 14C and 13C and with a trace amount of [3-3H]glucose. The glucose concentration in arterial plasma water averaged 6.4 +/- 1.0 mmol/L and in liver water 14.6 +/- 4.4 mmol/L (mean +/- SD).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8231848 TI - Osteoporosis and increased bone fractures in cerebrotendinous xanthomatosis. PMID- 8231849 TI - Tracer and nontracer studies yield similar conclusions. PMID- 8231850 TI - Hints on using the Guide to Techniques in Mouse Development. PMID- 8231851 TI - Recovery, capacitation, acrosome reaction, and fractionation of sperm. PMID- 8231852 TI - Strategies and methods for evaluating acrosome reaction. PMID- 8231853 TI - Culture of preimplantation embryos. PMID- 8231854 TI - Anesthesia and perioperative care. PMID- 8231855 TI - Isolation and culture of whole postimplantation embryos and germ layer derivatives. PMID- 8231856 TI - Histogenetic potency of embryonic tissues in ectopic sites. PMID- 8231857 TI - Micromanipulation of gametes and embryos. AB - The procedures described in this chapter are applicable to problems of oogenesis, fertilization, and early development. Many important procedures have not been described (e.g., pronuclear exchange). However, nearly every experimental manipulation thus far published for the mouse either directly involves the techniques included here or is closely related to them. Therefore, if all of the skills outlined in this chapter are successfully mastered, it should be possible to adapt that knowledge to most or all known experimental protocols. Successful establishment of micromanipulation is certain to have a significant and lasting positive impact on nearly any research program in mammalian development. PMID- 8231858 TI - In vitro capacitation and fertilization. PMID- 8231859 TI - In vitro fertilization. PMID- 8231860 TI - Quantitative analysis of specific messenger RNAs by hybridization using RNA probes. PMID- 8231861 TI - Quantitative analysis of specific messenger RNAs by ribonuclease protection. PMID- 8231862 TI - Recordkeeping and database analysis of breeding colonies. PMID- 8231863 TI - Detection of messenger RNA by in situ hybridization to tissue sections and whole mounts. PMID- 8231864 TI - Culture and manipulation of primordial germ cells. PMID- 8231865 TI - Detection of messenger RNA by in situ hybridization to postimplantation embryo whole mounts. PMID- 8231866 TI - Preparation of injection pipettes. PMID- 8231867 TI - Application of firefly luciferase to preimplantation development. PMID- 8231868 TI - Application of LacZ gene fusions to preimplantation development. PMID- 8231869 TI - Two-dimensional gel analysis of protein synthesis. PMID- 8231870 TI - One-dimensional gel analysis of histone synthesis. PMID- 8231871 TI - Whole-mount immunohistochemistry. PMID- 8231872 TI - Techniques for localization of specific molecules in oocytes and embryos. PMID- 8231873 TI - Isolation and culture of primordial germ cells. PMID- 8231874 TI - Construction of cDNA libraries from single cells. PMID- 8231875 TI - Construction and characterization of yeast artificial chromosome libraries from the mouse genome. PMID- 8231876 TI - Genome mapping and cloning of mutations using yeast artificial chromosomes. PMID- 8231877 TI - Cloning developmentally regulated gene families. PMID- 8231878 TI - Screening for novel pattern formation genes using gene trap approaches. PMID- 8231879 TI - Insertional mutagenesis by retroviruses and promoter traps in embryonic stem cells. PMID- 8231880 TI - Two-dimensional gel analysis of repetitive DNA families. PMID- 8231881 TI - Transplantation of nuclei to oocytes and embryos. PMID- 8231882 TI - Manipulations of genetic constitution by nuclear transplantation. PMID- 8231883 TI - Production of transgenic mice. PMID- 8231884 TI - Isolation and culture of oocytes. PMID- 8231885 TI - Factors influencing frequency production of transgenic mice. PMID- 8231886 TI - Surgical techniques in production of transgenic mice. PMID- 8231887 TI - Identification of transgenic mice. PMID- 8231888 TI - Derivation of embryonic stem cell lines. PMID- 8231889 TI - Production of chimeras between embryonic stem cells and embryos. PMID- 8231890 TI - Purification, culture, and fractionation of spermatogenic cells. PMID- 8231891 TI - Gene targeting in embryonic stem cells. PMID- 8231892 TI - Simple screening procedure to detect gene targeting events in embryonic stem cells. PMID- 8231893 TI - Manipulation of transgenes by site-specific recombination: use of Cre recombinase. PMID- 8231894 TI - Production of tetraploid embryos by electrofusion. PMID- 8231895 TI - Lineage analysis using retrovirus vectors. PMID- 8231896 TI - Allosteric regulation by sodium of the binding of [3H]cocaine and [3H]GBR 12935 to rat and bovine striata. AB - Sodium regulation of ligand binding to the dopamine transporter of rat and/or bovine striata was investigated using a filtration binding assay. In low Na+ phosphate or bicarbonate-buffered sucrose (300 mOsm), the tissue exhibited high affinity for [3H]cocaine which was reduced by the addition of Na+ in a dose dependent manner. However, [3H]GBR 12935 binding was insensitive to Na+ in these physiological buffers. Although binding of [3H]GBR 12935 was displaced by cocaine in a manner consistent with competitive displacement, a non-linear affinity shift of the displacement of [3H]GBR 12935 by cocaine suggests that the two ligands bind to distinct sites. Binding of both radioligands was suppressed when measured in sodium-free 50 nM Tris-sucrose and increased with the addition of Na+. Scatchard analysis indicated that Bmax for [3H]cocaine binding in Tris plus 120 mM NaCl reached the same level as in the physiological buffers. In Krebs-Ringer buffer with phosphate, bicarbonate or Tris, which contained 120 nM NaCl, both [3H]cocaine and [3H]WIN 35428 binding exhibited lower affinities than in Na(+) deficient phosphate buffer. It is suggested that the cation form of Tris binds to the dopamine transporter and that the Tris-receptor complex does not bind [3H]cocaine or [3H]GBR 12935. Na+ displaces Tris, forming a Na(+)-receptor complex which binds these ligands. Thus, it is suggested that the Na(+)-dependent binding of cocaine to the dopamine transporter is observed only in Tris. PMID- 8231897 TI - Solubilization of multilamellar liposomes in the presence of non-ionized drug. AB - The solubilization of multilamellar liposomes by metoprolol tartrate (MPL) has been studied as a function of pH, [MPL], [dimyristoylphosphatidylcholine (DMPC)], temperature and lipid composition. The solubilization of liposomes at 37 degrees C by 7.3 nM MPL occurred at different rates at different pH values. MPL completely solubilized by 7.2 mM DMPC liposomes after about 17 h at pH 12, but only a partial solubilization occurred at pH 10 and 11. Between pH 7 and 9 no change in turbidity was observed after 1 week. Addition of cholesterol (CHOL) to DMPC (2:1 mol) had very little effect on solubilization after 24 h, however with DMPC:CHOL (5:1 mol) the decrease in turbidity was observed after 24 h, even though solubilization was much less compared with that of DMPC alone. The rate of solubilization was decreased when dipalmitoylphosphatidylcholine liposomes were employed. Addition of dicetylphosphate (DCP) to DMPC liposomes reduced the rate of solubilization significantly. The solubilization of liposomes by 7.3 mM MPL as a function of [DMPC], indicated that the lower the liposome concentration the greater the effect on solubilization. It is concluded that MPL in the non-ionized form has a solubilizing effect on liposomes, and addition of CHOL or DCP to DMPC has a stabilizing effect against solubilization. PMID- 8231898 TI - Schistosoma mansoni: surface membrane isolation with lectin-coated beads. AB - Lectins from Lens culinaris and Arachis hypogaea immobilized on polyacrylamide beads were used for selective isolation of glycosylated surface membrane domains of adult Schistosoma mansoni worms, and the method was compared with the membrane isolation procedure developed with polycationic (Affi-Gel) beads. The lentil lectin proved to be suitable for interaction with surface membrane components: an increment in the specific activities of tegumental phosphohydrolases was observed in the bound fraction with respect to that observed in a total worm homogenate. A characteristic polypeptide pattern on gel electrophoresis was also seen, more restricted than that obtained with the bound Affi-Gel fraction. Immobilized peanut lectin was not successful as a method for isolating membrane material from the tegument of adult worms. Solubilization and dissociation of the lentil lectin bound enzyme markers was achieved after addition of detergent and competing sugars. Glycosylation of the solubilized enzymes was further confirmed by affinity chromatography with fresh lentil lectin-coated beads. These results, together with histochemical evidences, suggest that the active sites of some of these enzymes are located within or close to the cytoplasmic leaflet of the surface tegumental membranes, and allow us to propose a model for the double surface membrane complex where some proteins may be crossing the two bilayers. PMID- 8231899 TI - Synthesis of gap junction proteins and collagenases in the preimplantation rat uterus. AB - Synthesis of gap junction proteins (GJPs) and of collagenases in the rat uterus has been studied under two physiological conditions: various stages of the estrus cycle, and the early pregnancy period. The synthesis has been studied by incubating uterine horns in a short-term tissue culture medium containing radioactively-labeled amino acids, followed by a double antibody immunoprecipitation of the labeled proteins. After exposure of the media to either anti-collagenase IgG(s) or anti-GJPs IgG(s), the final immunoprecipitation was achieved with the use of goat anti-rabbit IgG. Collagenase(s) synthesis was found to reach the peak, during the estrus cycle, at the proestrus stage, while GJP synthesis reached the maximum during the estrus stage. In the preimplantation, pregnant, rat uterus the syntheses of both the proteins reached the respective peak activities on day 4 of pregnancy, about 24 h before the expected time of ovum implantation. A study of the literature reveals that this time coincides with a spurt in exposure of the progesterone dominated uterus to estradiol. PMID- 8231900 TI - Ca2+ permeability of rat parotid gland basolateral plasma membrane vesicles is modulated by membrane potential and extravesicular [Ca2+]. AB - This study examines the Ca2+ permeability of basolateral plasma membrane vesicles (BLMVs) isolated from the rat parotid gland by monitoring the rate of 45Ca2+ efflux from actively-loaded (via the Ca(2+)-ATPase) inside-out BLMVs. Ca2+ efflux from BLMVs into a K(+)-gluconate medium which hyperpolarizes the cytoplasmic side (i.e. outside) of the inside-out BLMVs resulted in a faster rate of Ca2+ efflux compared with a control medium containing N-methyl-D-glucamine (NMDG)-gluconate. Conversely, Ca2+ efflux into a medium which depolarizes the cytoplasmic side of the BLMVs (NMDG-chloride) resulted in slower rates of efflux compared with those observed with the control medium. This increased rate of 45Ca2+ efflux from the hyperpolarized BLMV was inhibited by 1 mM Ni2+, yielding a rate of efflux similar to the rate observed in depolarized BLMVs. The rate of Ca2+ efflux from BLMVs was affected by [Ca2+]o ([Ca2+] on the extravesicular, cytoplasmic side of the vesicle). When [Ca2+]o was kept > 200 nM during efflux, the rate of Ca2+ efflux from both hyper- and depolarized BLMVs was slow and relatively unresponsive to changes in [Ca2+]o, despite sizeable changes in the Ca2+ gradient across the BLMV. However, when [Ca2+]o was lowered < 200 nM, there was an abrupt increase in the rate of Ca2+ efflux from both hyper- and depolarized BLMVs. Additionally, when [Ca2+] was < 200 nM, the rate of Ca2+ efflux appeared to be more sensitive to driving force changes. These data suggest that Ca2+ permeability across the rat parotid gland basolateral plasma membrane is modulated by membrane potential and [Ca2+] on the cytoplasmic side. PMID- 8231901 TI - Swelling-stimulated passive potassium transport in camel erythrocytes: inhibitory effects of furosemide and sodium fluoride. AB - The inhibitory effects of furosemide, sodium fluoride, and age on volume dependent, ouabain-resistant K+ influx were investigated in camel red blood cells. Swelling of young camel erythrocytes hypotonically stimulates ouabain resistant potassium influx, a response that was lacking in old camel erythrocytes. The swelling-stimulated influx was partially inhibited by 1 mM furosemide and by 10 and 20 mM sodium fluoride. The inhibitory effect of furosemide was significantly increased if rubidium was added to the flux media. There was a significant correlation between potassium influx in normo- and hypotonic media which might indicate that the anion-dependent transport system operates, to some extent, to regulate cell volume. PMID- 8231902 TI - Effects of oophorectomy and hormone replacement therapy on pituitary-gonadal function. AB - The purpose of this study was to determine how oophorectomy and different hormone replacement therapy (HRT) regimens using low doses of medroxyprogesterone acetate (MPA, 2.5 mg/day) influence the pituitary-gonadal axis function. Ninety (90) women, who had had regular menses prior to surgery, completed a 1-year follow-up period. Patients were assigned to 5 groups. The first (n = 16) received 0.625 mg/day conjugated equine oestrogens (CEE) cyclically, the second (n = 20) 50 micrograms day transdermal oestradiol (E2) cyclically and the third (n = 15) 0.625 mg/day CEE continuously. These 3 groups also received 2.5 mg MPA sequentially for the last 12 days of HRT administration. The fourth group (n = 20) received 0.625 mg/day CEE and 2.5 mg/day of MPA continuously, while the fifth (n = 19) constituted a control group. After oophorectomy all patients showed increases in follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels, and decreases in those of E2, oestrone (E1), prolactin (PRL), sex-hormone binding globulin (SHBG), androstenedione (delta A4) and testosterone (T). No changes were detected in dehydroepiandrosterone sulphate (DHEA-S) levels. After HRT, decreases in FSH, LH and PRL levels and increases in those of E2, E1 and SHBG were observed, but no changes were seen in T, delta A4 or DHEA-S plasma levels. As the differences that were found cannot be attributed to the presence of ovaries, it is reasonable to assume that they were perhaps due to the treatment. All these changes, with the exception of a decrease in PRL levels, are therefore to be expected after HRT. PMID- 8231903 TI - Effects of oophorectomy and hormone replacement therapy on plasma lipids. AB - The aim of this study was to determine the effects on plasma lipids and lipoproteins of oophorectomy and various hormone replacement therapy (HRT) delivery systems using low doses of medroxyprogesterone acetate (MPA, 2.5 mg/day). A total of 90 women completed the 1-year follow-up period. Patients were randomly assigned to five groups. The first (n = 16) received 0.625 mg/day conjugated equine oestrogens (CEE) cyclically, the second (n = 20) 50 micrograms/day transdermal oestradiol cyclically and the third (n = 15) 0.625 mg/day CEE continuously. These three groups also received 2.5 mg MPA sequentially for the last 12 days of HRT administration. The fourth group (n = 20) received 0.625 mg/day CEE and 2.5 mg/day MPA continuously, while the fifth (n = 19) constituted a treatment-free control group. After oophorectomy patients showed increases in low-density lipoprotein (LDL), apolipoprotein B and the atherogenic index, whereas after HRT patients exhibited falls in plasma LDL, apolipoprotein B and the atherogenic index and increases in high-density lipoprotein (HDL) and apolipoprotein A1. No significant changes in total cholesterol were observed after surgery or treatment and decreased levels of triglycerides were detected only in the transdermal treatment group. PMID- 8231904 TI - Effects and acceptability of a new 17 beta-oestradiol-releasing vaginal ring in the treatment of postmenopausal complaints. AB - The treatment of postmenopausal symptoms was studied in 26 healthy women using a new synthetic rubber (Kraton D 2109) vaginal ring containing 53 mg of 17 beta oestradiol. All women were postmenopausal, at least 6 months after the last vaginal bleeding and suffering from daily hot flushes. The study was conducted in a double-blind placebo-controlled intrapatient cross-over fashion, and the study period was 6 months. The rings used give an initial in vitro release rate of 0.4 mg/E2 per day. The in vitro release rate decreases to about 0.2 mg/day after 20 days and levels off asymptotically to about 0.1 mg/day after 50 days. Serum E2 levels equivalent to the follicular phase of the normal menstrual cycle were measured after 1 month's use, and serum E2 level stayed above postmenopausal levels throughout the study period. FSH was suppressed during use of the E2 releasing vaginal ring, while LH showed no statistically significant suppression in continued use. Postmenopausal complaints were recorded by Visual Analogue Scales (VAS) as judged by both the patient and the examining doctor; all complaints had favourable outcomes during use of the E2-releasing vaginal ring without deterioration of symptoms during use of the placebo ring. No serious side effects were encountered, and the possibility of managing all postmenopausal complaints with intravaginal oestrogen treatment is discussed. PMID- 8231905 TI - Sexual behaviour and attitudes of Danish middle-aged men-methodological considerations. AB - A Danish cohort-investigation of health risk-factors (started in 1976) included descriptions of the sexuality of the participants. This paper examines the validity/representativity of the results of the sexological investigation of the male participants, when in 1987 they were 51 years old. Remaining results will be published in separate articles about each main theme. The participants, 439 men, completed a questionnaire concerning sexuality (behaviour, attitudes and experience) and had their serum-LH and serum-free testosterone measured. Thereafter 100 of the 439 men were interviewed. The present examination shows that the participants are representative of same-aged men from all over Denmark concerning the sexological results. The answer ratio of the questionnaire was 70 95% and the non-responders of the questionnaire gave different interview answers than the rest. Complemented with their answers (answer ratio of nearly 100%) the interview results are more representative than the results of the questionnaire. The interview information is in this study considered more valid than the information obtained by the questionnaire--because the former more frequently than the latter contain answers embarrassing to the participants (e.g. high frequency of masturbation) or difficult for them to recall. The interview thereby adds vital information to that of the questionnaire. PMID- 8231906 TI - Relationships between attitude towards menopause, well-being and medical attention among women aged 45-60 years. AB - This study aims to answer the following questions: (i) what is the attitude of women in the climacteric years towards menopause, (ii) what is the association between attitude towards menopause and well-being and (iii) to what extent is medical attention determined by both well-being and attitude towards menopause. All 2729 women aged 45-60 years living in a suburb of Rotterdam were sent a questionnaire, of these 1947 (71.3%) were returned. Attitude was measured on a 5 point rating scale using 28 items that have been used in other studies. Well being was measured by the Inventory of Subjective Health and three subscales of the Sickness Impact Profile. Medical attention was measured by asking the women whether they were currently being treated by a general practitioner or specialist. Results show that three clusters of attitudes towards menopause exist: two clusters encompasses items reflecting attitudes towards disadvantages and advantages of the menopause, one cluster encompasses items reflecting attitudes towards (medical) treatment of the menopause. On the whole, women answer neutrally to items relating menopause with the disadvantages and tend to agree with items relating menopause with the advantages. The women slightly agree, premenopausal women more than others, with items that are in favour of treatment of menopausal complaints. Agreement with items on the disadvantage cluster is moderately associated with a low level of well-being, whereas agreement with items on the advantage cluster is slightly associated with a high level of well-being; the treatment cluster is not associated with well-being. Both well-being and agreement with items on the treatment cluster are statistically significantly associated with medical attention. Apart from these variables, the woman's ideas about treatment are also related to medical attention. PMID- 8231907 TI - Osteoporosis risk factors, gynaecological history and fractures in perimenopausal women--the results of the baseline postal enquiry of the Kuopio Osteoporosis Risk Factor and Prevention Study. AB - The Kuopio Osteoporosis Risk Factor and Prevention (OSTPRE) Study examines the risk factors of osteoporosis, the relationship of risk factors to bone density and fractures, as well as the possibility to prevent bone loss by administering certain hormones. The baseline postal enquiry in 1989 was sent to all the 14,220 women aged 47-56 years residing in the Kuopio Province, Finland. The questionnaire included questions about their gynaecological history, physical exercise and smoking habits, calcium intake, body weight and height, history of bone fractures, health disorders, their current and previous use of drugs, as well as their willingness to participate in bone densitometry and in a clinical hormone trial. The response rate was 92.8%. In all, 56% reported some previous use of female hormones. Strong contraindications for oestrogen replacement therapy were found in 9.3% of the women. Almost half of the respondents reported lack of regular physical exercise, 11.9% were smokers, and 17.0% reported a calcium intake from milk products of less than 500 mg daily. The incidence of fractures increased steadily with age. The incidence of premenopausal fractures within the last 10 years was 7.65 per 1000 person/years and that of postmenopausal fractures was 17.40 per 1000 person/years (P = 0.000). The effect of menopause on fracture incidence was stronger than the effect of a 5-year age increase. Of the respondents, 84.4% were willing to participate in bone densitometry and 68.3% for long-term prevention of osteoporosis with oestrogen. PMID- 8231908 TI - The public health breast and cervical cancer program. Our role as physicians. AB - Through a federally funded breast and cervical cancer control program, women who have breast or cervical cancer should be identified at earlier stages of their disease. Working together, community physicians and local health departments can ensure that the highest quality breast and cervical cancer control services are available to all women in their communities. This public/private partnership will help ensure the survival of these crucial members of our families and communities -our mothers, grandmothers, wives, sisters, daughters, friends, and co-workers. For information about the location of participating local health departments, please call the Michigan Department of Public Health Mammogram Hotline at 1-800 922-MAMM (1-800-922-6266). For media materials or for more information about becoming involved as a provider of services for community breast and cervical cancer screening programs, call (517) 335-9161. All women are at risk of developing breast and cervical cancer. Please do what you can to encourage all women to obtain screening according to the schedule recommended by the American Cancer Society and the National Cancer Institute. And please cooperate with your local health department when your community initiates this program. References available upon request. PMID- 8231909 TI - Good health can come from education, prevention. MSMS president-elect targets youth. PMID- 8231910 TI - Cancer incidence among Michigan residents 1989. PMID- 8231911 TI - Kent County mini-internship program helps demystify medicine. PMID- 8231912 TI - Two Michigan physicians establish the American Russian medical exchange. PMID- 8231913 TI - Douglas A. Mack, MD, MPH. PMID- 8231914 TI - Health Education Foundation. Physician care reaches into the community. PMID- 8231916 TI - Information sought on human health effects of nonionizing electromagnetic radiation. PMID- 8231915 TI - Clinical laboratory improvement amendments (CLIA) of 1988. Here are answers to your most frequently asked questions. PMID- 8231917 TI - Certificate of Need laws increase medical care costs. PMID- 8231918 TI - Bulimia nervosa linked to brain hormone, U-M study indicates. PMID- 8231919 TI - MDPH study of anti-smoking/AIDS prevention efforts released. PMID- 8231920 TI - AMA sets health reform strategies for 1993. PMID- 8231921 TI - Medical study targets leading cause of blindness in elderly. PMID- 8231922 TI - State releases 1993 fish consumption advisory. PMID- 8231923 TI - U-M researchers unfold new data on beta-carotene. PMID- 8231924 TI - Physicians, hospitals, and managed competition. The beginning of a new era in medicine. PMID- 8231925 TI - New MSMS service will help "level the playing field" in contract negotiations. PMID- 8231926 TI - PHOs enhanced health care delivery through physician-hospital cooperation. AB - In light of the many trends that have been building over the past decade and the current pressures on the health care system for reform, change is inevitable in the way medical and other health care services are delivered and financed. Regardless of the precise nature of those changes, increased cooperative effort between physicians and hospitals in providing health care at the community level will be critical in ensuring the future success of the health care system. PHOs represent one potential approach to increased health care integration at the local level which can offer significant advantages to patients, physicians, and hospitals. Although PHOs are not a panacea for the ills facing the U.S. health care system or for deteriorating physician-hospital relations, they can be a solid building block for a stronger health care system and for improved, cooperative relations between health care providers. PMID- 8231927 TI - Managed competition. Some concerns for physicians. PMID- 8231928 TI - Compassionate alternatives for the terminally ill. PMID- 8231929 TI - Health expenditures: considering all the costs. PMID- 8231930 TI - Michigan drug utilization review and OBRA 90. AB - The DUR Board's role in the Medicaid DUR process is significant. With the requirements for membership on the DUR Board and the responsibilities directed by OBRA 90, the standards will ensure that they reflect the accepted medical practice and are clinically relevant to the geographical area represented. The DUR Board has the potential to improve drug utilization review and ultimately improve the quality of care. The approval of criteria for both patient retrospective and prospective DUR and the addition of education intervention will help us to improve the quality of care. DSS encourages your input and participation on the DUR regional committees. There are currently three committees that meet monthly and are located in: Detroit, Ann Arbor, Lansing. PMID- 8231931 TI - Informed consent should be required before HIV testing. PMID- 8231932 TI - Current HIV/AIDS testing policies barriers to care. PMID- 8231933 TI - Update on pension plan protection. PMID- 8231934 TI - Physician profile appreciated. PMID- 8231935 TI - Results of the MSMS Survey on Practice Characteristics. PMID- 8231936 TI - Good medical records key to prevention, defense against malpractice claims. PMID- 8231937 TI - How to cope with the stresses of practice. PMID- 8231938 TI - Physicians are people too. PMID- 8231939 TI - MSMS adopts policy statement on physician-assisted suicide. PMID- 8231940 TI - MSMS Group Insurance Trust invites member participation. PMID- 8231941 TI - MSMS reimbursement roundup. PMID- 8231942 TI - Michigan physicians doing a "good job" of utilizing Medicare's new E & M codes. PMID- 8231943 TI - Articles on informed consent before HIV testing raise more questions than answers. PMID- 8231944 TI - How I survive health care of the 90s with my computer. PMID- 8231945 TI - Electronic billing: it's more than just a coming attraction. Here's how to take the first step. PMID- 8231946 TI - John W. Finn, MD. "It is a privilege to care for the dying". PMID- 8231947 TI - It's time to focus on finances. PMID- 8231948 TI - National Coalition of Physicians Against Family Violence. More than 180 Michigan physicians, alliance members, and professionals are now involved. PMID- 8231949 TI - The Michigan Limited Liability Company. A new structure for medical practice. PMID- 8231950 TI - Managed care: just what is it anyway? PMID- 8231951 TI - Medical records--Part Two. PMID- 8231952 TI - MSMS AIDS provider education update. PMID- 8231953 TI - MSMS: preparing Michigan physicians for change. PMID- 8231954 TI - MSMS physician organizations committee: helping Michigan physicians organize for the future. PMID- 8231955 TI - Physician organizations and physician-hospital organizations: models for managing change. AB - The health care environment is expected to undergo significant change in the coming years. The continued growth of managed care, in particular will exert a profound impact on medical practice. This trend will be accelerated if a managed competition approach to health system reform is adopted at the federal level. As physicians continue to consider possible strategies by which to adapt to the changing practice environment, two options may merit special consideration: first, integration with other physicians, particularly through formation of a medical group and, second, vertical integration with a hospital, through formation of a PHO. Although PHOs are a relatively recent phenomenon, they represent a potentially powerful strategy by which physicians and hospitals can combine forces to take a leadership role in managed care contracting. Through careful planning, development of a sound legal structure and efficient governance process, and implementation of effective criteria for participation in the PHO, it is possible to further the goals of the physician and hospital participants, while meeting the needs of payers. PMID- 8231956 TI - Driving the nitroglycerin truck. PMID- 8231957 TI - MSMS spring PO/PHO conference provided a wealth of information for Michigan physicians. PMID- 8231958 TI - Immunizations: physicians must act now to ensure all our children are immunized. PMID- 8231959 TI - Effect of Clostridium perfringens epsilon toxin on rat isolated aorta. AB - The effect of Clostridium perfringens epsilon toxin on rat isolated aorta was investigated. The toxin caused contraction of the isolated aorta in a dose dependent manner. The toxin induced no contraction of the isolated aorta in low Na medium and of the tissue stored at 4 C for 7 days. However, tetrodotoxin (TTX) had no effect on the toxin-induced contraction. The toxin-induced contraction was significantly inhibited by phentolamine and prazosin, but did not by atropine, mecamylamine, chlorpheniramine and methysergide. These data suggest that the toxin-caused contraction is mediated through nervous system in rat isolated aorta. PMID- 8231960 TI - Cloning and sequencing of two new Verotoxin 2 variant genes of Escherichia coli isolated from cases of human and bovine diarrhea. AB - We cloned and sequenced two new Verotoxin 2 (VT2) variant genes: one from an Escherichia coli strain from a case of bovine diarrhea and the other from an E. coli strain from a patient with diarrhea. The nucleotide and amino acid sequences of these two genes were highly homologous with, but distinct from those of the VT2, VT2vha, VT2vhb, SLT-IIv (VT2vp1) and SLT-IIva (VT2vp2) genes. Their nucleotide sequences were much more closely homologous to that of VT2vh than to that of VT2vp. Search for these two new genes in other Verocytotoxin-producing E. coli strains resulted in the isolation of 2 strains carrying one of the new VT2 variant genes, one strain from Tokyo and the other from Canada. PMID- 8231961 TI - A positive correlation between the precursor frequency of cytotoxic lymphocytes to autologous Epstein-Barr virus-transformed B cells and antibody titer level against Epstein-Barr virus-associated nuclear antigen in healthy seropositive individuals. AB - A limiting dilution analysis was established to determine the precursor frequency (PF) of cytotoxic lymphocytes against autologous B cells transformed with the Epstein-Barr virus (EBV). This method was found to detect mainly self-restricted T-cell activity and little non-self-restricted cytotoxicity. The mean PF in 21 healthy EBV-seropositive persons was 1.4 x 10(-3) (range: 0.03 x 10(-3) to 8.7 x 10(-3)) for peripheral blood mononuclear cells, whereas 4 samples of mononuclear cells obtained from umbilical cord blood had PFs below 0.007 x 10(-3). A positive correlation was observed between the PF and serum antibody titers against EBV associated nuclear antigen among the seropositive persons. PMID- 8231962 TI - The nucleotide sequence and genome structure of mung bean yellow mosaic geminivirus. AB - Complete nucleotide sequences of the infectious cloned DNA components (DNA 1 and DNA 2) of mung bean yellow mosaic virus (MYMV) were determined. MYMV DNA 1 and DNA 2 consists of 2,723 and 2,675 nucleotides respectively. DNA 1 and DNA 2 have little sequence similarity except for a region of approximately 200 bases which is almost identical in the two molecules. Analysis of open reading frames revealed nine potential coding regions for proteins of mol. wt. > 10,000, six in DNA 1 and three in DNA 2. The nucleotide sequence of MYMV DNA was compared with that of bean golden mosaic virus (BGMV), tomato golden mosaic virus (TGMV) and African cassava mosaic virus (ACMV). The 200-base region common to the two DNAs of each virus had little sequence similarity, except for a highly conserved 33-36 base sequence potentially capable of forming a stable hairpin structure. The potential coding regions in the MYMV DNAs had counterparts in the BGMV, TGMV and ACMV, suggesting an overall similarity in genome organization, except for absence of 1L3 in MYMV DNA 1. The most highly conserved ORFs, MYMV 1R1, BGMV 1R1, TGMV 1R1 and ACMV 1R1, are the putative genes for the coat proteins of MYMV, BGMV, TGMV and ACMV, respectively. MYMV 1L1 has also a high degree of sequence similarity with BGMV 1L1, TGMV 1L1 and ACMV 1L1. PMID- 8231963 TI - Suppressive activity of splenic macrophages induced by transparent and opaque colonial variants of Mycobacterium avium complex against lymphoproliferative response. AB - Splenic macrophages (M phi s) induced by the two colonial variants of Mycobacterium avium complex, SmT variant (smooth, transparent, flat) and SmD variant (smooth, opaque, domeshaped), were studied for their suppressive activity against mitogenic response of splenic T cells to concanavalin A (ConA). The SmT variant is much more virulent than the SmD variant and causes more severe infection in host mice. Moreover, suppressed state of ConA mitogenic response of host spleen cells persisted much longer in SmT-infected mice than in SmD-infected ones. However, the suppressive activity of SmT-induced M phi s, induced around 2 weeks after infection, against lymphoproliferative response was significantly weaker than SmD-induced M phi s. In contrast, the former M phi s showed somewhat higher active oxygen-producing ability than the latter, as based on the chemiluminescence. These findings indicate that the immunosuppressive M phi s transiently induced in the early phase of M. intracellulare infection hardly affect the host resistance to the organisms and, thus, play relatively minor role in the progression of the infection. PMID- 8231964 TI - An occurrence of diarrheal cases associated with group C rotavirus in adults. AB - Six of the 23 college students who joined a group trip in February of 1991 developed acute nonbacterial gastroenteritis with severe diarrhea. The causal agent was identified as group C rotaviruses by electron microscopy (EM), immune EM (IEM) and the molecular examinations including polyacrylamide gel electrophoresis (PAGE) and polymerase chain reaction (PCR) on virus particles detected in the extract of watery fecal specimens of the patients. The patients positive for virus isolation showed significant increase in IEM antibody to the isolated virus in their paired sera. These findings suggest that the group C rotavirus is an important etiological agent of diarrhea and may also cause serious food-borne diarrheal disease in adults. PMID- 8231965 TI - Mechanism of inhibition of DNA synthesis by 1-beta-D-arabinofuranosyl-E-5-(2 bromovinyl)uracil. AB - The mechanism of the inhibitory action of 1-beta-D-arabinofuranosyl-E-5-(2 bromovinyl) uracil triphosphate (BV-araUTP) on DNA synthesis by Escherichia coli DNA polymerase I Klenow fragment was studied. Acting as a chain terminator, BV araUTP inhibited DNA synthesis by Klenow fragment more effectively than 2',3' dideoxythymidine triphosphate (ddTTP). However, the incorporation sites of BV araU monophosphate were restricted at consecutive dTMP sequence whereas ddTMP was incorporated at every dTMP site. PMID- 8231966 TI - Chlamydia pecorum--the fourth species of genus Chlamydia. PMID- 8231967 TI - Changes in the susceptibility of 12-O-tetradecanoylphorbol 13-acetate (TPA) treated HL-60 cells to staphylococcal leukocidin. AB - Susceptibility of human promyelocytic leukemia HL-60 cells to staphylococcal leukocidin following treatment of cells with 12-O-tetradecanoylphorbol 13-acetate (TPA) was examined. TPA treatment for 6 hr rendered the cells very resistant transiently to leukocidin. There was no change in binding of leukocidin to the cells, but leukocidin-induced 45CaCl2 influx, phospholipase A2 and C activities were inhibited. Further incubation with TPA rendered the cells sensitive again and then more sensitive than original HL-60 cells following increase of the binding, and leukocidin-induced activities described above appeared again. Those cells treated with TPA for more than 18 hr started to differentiate to macrophages morphologically and functionally. These data suggest that the differentiated cells were more sensitive than original HL-60 cells because of increased binding of leukocidin and that treatment of TPA for 6 hr may transiently impair the signal transduction system of leukocidin after binding of leukocidin to the specific receptor of the cell membrane. Using these TPA-treated cells, it was shown in this report that calcium influx, phospholipase A2 and C activities were important to induce cytotoxic action of leukocidin after binding of leukocidin to specific receptors on the cells. PMID- 8231968 TI - Detection of various variant verotoxin genes in Escherichia coli by polymerase chain reaction. AB - We constructed common primers for the polymerase chain reaction to detect the genes for various Verotoxins reported, that is, VT1 (or SLT-I), VT2 (or SLT-II), VT2vha, VT2vhb, SLT-IIv (or VT2vp1, VTe) and SLT-IIva (or VT2vp2). A total of 80 Verocytotoxin-producing Escherichia coli strains isolated from humans, domestic animals and meats gave a positive result by PCR with the designed common primers. Digestion by restriction endonucleases BglII and EcoT14I of the amplicon of the VT2vp2 gene gave specific bands of the expected sizes, but not of the amplicons of other VT genes, suggesting a possible method for identification of the VT2vp2 gene. Application of the PCR with the designed primers in diagnostic and epidemiological studies on VTEC infection is also discussed. PMID- 8231969 TI - Polymorphism of crystals of Salmonella minnesota Re and Ra lipopolysaccharides. AB - Salmonella minnesota Re and Ra lipopolysaccharides (LPSs) formed three dimensional crystals when they were precipitated by the addition of 2 volumes of 95% ethanol containing 375 mM MgCl2 and incubated in 70% ethanol containing 250 mM MgCl2 at 4 C. Besides typical shapes of crystals, hexagonal plates and solid columns, which were already reported (J. Bacteriol. 172: 1516-1528 (1990)), the LPSs thus treated formed crystals possessing various shapes such as square or rectangular plate, lozenge plate, discoid, and truncated hexangular pyramid forms. Electron diffraction patterns from all these crystals except square or rectangular plate crystals obtained by electron irradiation from the direction perpendicular to the basal plane were essentially the same as those from hexagonal plate crystals, indicating that they consist of hexagonal lattices with the lattice constant of 4.62 A. From these results as well as the results of electron microscopic observations of these crystals, it was concluded that all these crystals except square or rectangular plate crystals are composed of hexagonal plate sheets as the basic structural units. Square or rectangular crystals were assumed to correspond to the (1011) planes of solid hexagonal column crystals. PMID- 8231970 TI - Induction of acute arthritis in mice by peptidoglycan derived from gram-positive bacteria and its possible role in cytokine production. AB - The activities of a water-soluble peptidoglycan fragment derived from Staphylococcus epidermidis (SEPS) were examined as to their role in proliferation of spleen mononuclear cells (SMNC) from various strains of mice, the production of cytokines in vitro, and the induction of an inflammatory reaction in vivo. The proliferation of SMNC from C3H/HeN, C57BL/6, AKR, DBA/2, and ddY mice in reaction to SEPS in vitro showed a peak on day 3 and was greater than that of SMNC from BALB/c mice. The cells of SMNC from C3H/HeN mice responsive to SEPS were indicated to be mainly macrophages. A time kinetics experiment showed a coincidence in the proliferation of SMNC in reaction to SEPS and the detection of colony-stimulating factor (CSF) activity. Interleukin 2 (IL-2) activity was not detected during the incubation periods. When SEPS was administered to mice, much stronger mRNA transcripts of granulocyte-macrophage (GM)-CSF were detected in the lungs of C3H/HeN mice than in BALB/c mice. On the other hand, the amounts of IL-1 and PGE2 produced by SMNC of BALB/c mice stimulated by SEPS were greater than those produced in C3H/HeN mice. SEPS was confirmed to induce arthritis in BALB/c mice, but not in C3H/HeN mice. Our findings suggest that the production of GM-CSF is involved in the in vitro proliferation of SMNC in reaction to SEPS and that along with IL-1 and PGE2 production, contributes to the inflammation by SEPS in vivo. PMID- 8231971 TI - Enhancement by recombinant human interleukin 2 of host resistance to Toxoplasma gondii infection in pregnant mice. AB - The lymphokine production by pregnant mice infected with a lethal dose of Toxoplasma gondii was evaluated in comparison with that by virgin mice infected with a sublethal dose of this protozoan parasite. Splenocytes taken from mice before and on the day after infection produced considerable amounts of IL-2 in response to concanavalin A (Con A) stimulation, but the titers rapidly declined in both pregnant and virgin mice as infection progressed. A trace amount or undetectable level of IL-2 was produced by splenocytes from acutely infected mice when stimulated with Toxoplasma lysate antigen (TLA). In contrast to the kinetics of IL-2 production, the levels of IFN-gamma produced by splenocytes cultured with Con A or TLA increased steadily in the later stage of infection in both pregnant and virgin mice. Thus, the response to Con A or TLA of splenocytes to produce IL 2 and IFN-gamma differed strikingly in acute toxoplasmosis in mice. The administration of rHuIL-2 resulted in a significant decrease in the mortality of pregnant mice infected with a lethal dose of Toxoplasma. The combination of rHuIL 2 and rMuIFN-gamma increased the survival rate slightly but not significantly compared with pregnant mice receiving either rHuIL-2 or rMuIFN-gamma. Moreover, exogenously administered rHuIL-2 enhanced the production of both IFN-alpha and IFN-gamma in the bloodstreams of pregnant mice, in accordance with the decreased mortality. These results indicate that IL-2 may play a significant role in modulating the host defense against Toxoplasma infection in pregnant mice. PMID- 8231972 TI - A case that biomedicine is unduly negative to radical hypotheses and to theorizing--evidence based on cardiology's reaction to the spasm of resistance vessel concept and on the nature of scientific research. AB - This communication will attempt to make a case that biomedicine is unduly negative to radical hypotheses and to theorizing. Evidence will be based on a proposed undue negativity by cardiology to a radical hypothesis I first described two and a half decades ago--the spasm of resistance vessel (S-RV) concept of ischemic heart disease (IHD). The theory is regarded as an alternate paradigm and deals with basic pathogenetic mechanisms of IHD, the most significant disorder of Western civilization. The concept, if valid, might help in reducing the impact of this disorder, and I believe that the evidence for the theory and the importance of IHD support a more open minded attitude toward the idea. Cardiology's negativity is attributed to the nature of research; the most important factor is considered to be the Kuhnian negativity of scientific communities to hypotheses which are destructive of conventional wisdom, and a second factor is the special nature of biomedical research. Biomedicine is regarded as special because a low level of specific information about complex biomedical processes has fostered an essentially total study-based approach. Such an approach is assumed to have resulted in biomedicine's use of induction as 'the' method of scientific inquiry, and prompted negativity towards the hypothetico-deductive method used to develop and test the theory. Also, the study-based nature of biomedicine appears to have fostered an intuitive reliance on only newly performed studies to test hypotheses, which led to ignoring evidence for the concept derived from known information about IHD. Biomedicine is also regarded as special because its infrequent use of paradigm-change has resulted in unfamiliarity with this method, and because the practical method of training in biomedical research has worsened the general unfamiliarity of scientists with theoretical aspects of science. Because of these factors, the S-RV concept has not yet been properly evaluated--a quarter of a century after it was first created. PMID- 8231973 TI - Evidence in support of the spasm of resistance vessel concept of ischemic heart disease: an update in 1993. AB - The goal of this communication is to provide more evidence for the ischemic heart disease (IHD) component of the spasm of resistance vessel (S-RV) concept of IHD and other ischemic diseases. The S-RV concept of IHD is considered to be an alternate paradigm which challenges the accepted understanding of this disorder. The theory asserts that primary S-RV directly induces symptoms in IHD, and this position is in opposition to the accepted view that symptoms are induced directly by primary occlusions of epicardial arteries by coronary artery disease, spasm, and thromboses. The theory, if valid, should be useful in reducing the impact of IHD, as it generally is accepted that the correct appreciation of basic pathophysiological mechanisms helps ensure the most appropriate prevention and treatment of disease. PMID- 8231974 TI - Electromagnetic fields: a cancer promoter? AB - This paper discusses some of the current questions observed in updated research analyzing the association of electromagnetic fields (EMF) exposure and cancer. It reveals the predominant efforts toward obtaining more accurate levels of EMF exposure as a crucial key to gain knowledge about that association. A general picture of a molecular biological model of cancer is presented, showing that EMF presents certain compatible characteristics of a cancer promoter. Further, we raise the hypothesis that EMF action is probably linked with other unknown initiator agents, rather than acting alone as presently assumed in epidemiologic studies. Finally this paper reinforces the importance of strong descriptive analysis of potential factors acting together on EMF and cancer association. PMID- 8231975 TI - EMF-cancer link: the ferritin hypothesis. AB - Biological studies have not yet provided direct evidence for a link between electromagnetic fields (EMFs) and cancer. The hypotheses I present here describes ferritin as a direct pathway for the EMF-cancer link, which is supported by previous studies: (a) Ferritin plays a very important role in its associated cancers. (b) Iron is the principal regulator of ferritin synthesis and ferritin gene expression is induced by signals including hormones and cAMP. (c) Preliminary biological studies have demonstrated that EMFs induce some cellular effects including changes in hormone levels, alterations in cell membranes, variations of intracellular cAMP and even the modification of protein synthesis. So, it is reasonable to link the biological effects of EMFs with ferritin gene expression and ferritin synthesis which are mainly regulated by iron, hormones and cAMP and eventually the EMF-cancer link. PMID- 8231976 TI - The possible relationship between mercury from dental amalgam and diseases. I: Effects within the oral cavity. AB - Mercury is released from dental amalgams, and therefore it is necessary to consider the biological and clinical consequences of such exposure. Intraorally, it would appear as though mercury can cause hypersensitivity/toxic reactions resulting in lichen planus lesions, and may play a major role in the pathogenesis of gingivitis, periodontitis and periodontal disease. PMID- 8231977 TI - Carbon dioxide level as a migraine threshold factor: hypothesis and possible solutions. AB - Cerebral blood flow is highly sensitive to carbon dioxide (CO2) level changes. The existence of a migraine threshold to the CO2 air concentration is hypothesized. Decreasing the CO2 concentration in living and working environments may inhibit attacks which start during sleep, and prolong the migraine-free period. Carbon dioxide absorption characteristics by green plants, or by artificial means, are evaluated as non-drug long-term therapy methods. PMID- 8231978 TI - Genomic imprinting: a proposed explanation for the different behaviours of testicular and ovarian germ cell tumors. AB - Gonadal germ cell tumors in males are malignant in greater than 98% of cases while their ovarian counterparts are benign in 97% of cases. It has recently become clear that the maternal and paternal genomic contributions to the fertilized egg, provided by the haploid germ cells, have different and, to some extent, complementary effects on the developing embryo. This phenomenon, termed genomic imprinting, is a result of epigenetic modification of the genomes of male and female germ cells that has occurred in a sex-specific way during gametogenesis. There is a balance between the paternal and maternal effects in a normal embryo; results to date suggest that the paternal imprint promotes growth while the maternal imprint acts to limit potentially harmful overgrowth. It is proposed that differences in clinical behaviour between ovarian and testicular germ cell tumors can be explained by differences in the normal pattern of genomic imprinting of female and male germ cells. PMID- 8231979 TI - Acid/base may be more variable than previously thought. AB - Organ systems in the human body may not be homogeneous in regard to pH. Possibly specific organ systems under certain conditions maintain a pH level significantly different than the systemic pH. PMID- 8231980 TI - Dermatomyositis/polymyositis associated with internal malignancy: a consequence of how neoplasms alter generalized extracellular matrix in the host. AB - The mechanism of dermatomyositis/polymyositis (DM/PM) is unknown. There are multiple probable trigger mechanisms. Internal malignancy is a specific trigger for some cases of DM/PM. It is known that there are marked changes in the extracellular matrix around tumors and that various fractions of depolymerized glycosaminoglycans enter the circulation. Circulating ECM fractions are known to incorporate in the extracellular matrix (ECM). Skin changes in DM include mucin formation, edema, and atrophy. These are not post inflammatory changes. These changes could be a consequence of the tumor's effect in the generalized ECM. Many factors influence ECM. Infections are another probable trigger for DM/PM. Infections induce extracellular matrix changes. It is likely that a role for infectious agents in DM/PM will eventually be defined. The drug D-penicillamine is also a trigger for DM/PM. Understanding the mechanism of any one trigger might aid in helping define other triggers. Four other cutaneous signs of internal malignancy can be explained by the mechanism used to explain DM/PM. PMID- 8231981 TI - Actinic prurigo. The possible etiologic role of an amino acid in the diet. AB - Actinic prurigo is a chronic skin disease commonly associated with cheilitis and conjunctivitis. This peculiar disorder, with ethnic, genetic, environmental, clinical and histological features, represents an abnormal reaction to sunlight. It occurs in American Indians of Canada, United States and most frequently in Latin America where it involves not only indigents but also the mixed ancestry (Mestizo) population. In analogy to what is well known about pellagra, a disease also with solar influence and a deficient intake of tryptophan or niacin in diet, it is our hypothesis that actinic prurigo patients develop their disease due to, among other factors, a diet poor in proteins. PMID- 8231982 TI - A reconsideration of the measles-adherent leukocyte phenomenon and multiple sclerosis. AB - Some studies have shown that more mononuclear leukocytes (MNLs) from multiple sclerosis (MS) patients adhere to cells persistently infected (PI) with measles virus than do MNLs from controls. Some studies have shown no difference in adherence. It is suggested that the conflict between studies is merely technical. The net effect of this conflict has been to obscure the possibility of cell surface differences which underlie this adherence phenomenon and discourage further investigation of them. Given the current interest in the molecular basis of cell-cell adhesion and the new methods and reagents available to study it, the problem of identifying cell surface differences between normal and MS leukocytes can now be more readily approached. An investigation of the MS MNL cell surface using these new methods is suggested. PMID- 8231983 TI - A set-theoretical definition of the physical memories of stimuli. AB - Presumably, the memory of a stimulus exists as a difference in a system due to that stimulus, compared to what the system would have been had the stimulus never occurred. Thus every stimulus, to be remembered, must define with the system a set of phenomena perfectly divergent from those of other stimuli. Therefore there exist no subsets of physical phenomena common to memories of different stimuli, even those the experimenter classifies as equivalent. It is concluded that human awareness of groups of stimuli requires a dualistic explanation. PMID- 8231984 TI - A model of nerve regeneration in diabetic neuropathy. AB - The continued difficulty in discovering the cause of diabetic neuropathy may be due to the fact that researchers persist in looking for a factor associated with diabetes mellitus which damages nerves. The hypothesis that diabetic neuropathy is in fact related to some diabetes-related factor which slows nerve regeneration seems to fit the observed facts better. A model of nerve regeneration in diabetic neuropathy is presented, which facilitates understanding of recently published data on nerve regeneration in human diabetics. PMID- 8231985 TI - The epidemiology of neural tube defects. A mathematical model. AB - The incidence of neural tube defects (NTDs) shows a seasonal variation; and incidence as well as female/male ratio show a relation with latitude. The interrelation of these phenomena is presented as a mathematical model, which has a 'predictive' value, and is an instrument in estimating the local gene frequency. The model offers a simple explanation of the conundrum why the double X-chromosome has a variable influence on the sex ratio. UV light and the herpes virus fit in this model as related causative factors of NTDs. PMID- 8231986 TI - Mechanisms for methylation-mediated gene silencing and aging. AB - Gene silencing is often mediated by CpG methylation of key protein binding sites within gene regulatory sequences (GRSs). An aging mechanism is proposed based on this gene-silencing phenomenon whereby accumulation over time of methylation within GRSs contributes to cellular senescence. The proposed molecular mechanism for age-related gene silencing is the spreading of methylation through the regulatory sequences of genes resulting in progressive reduction of gene transcription. There is considerable experimental evidence for methylation spreading and its role in gene silencing, but the mechanism responsible for this process has not been elucidated. A four-step mechanism is proposed whereby an original methylation occurs, methyltransferase (MTase) molecules progressively move 5' to 3' from this site, neighboring CpG dinucleotides become methylated, and diminished gene expression ensues. Over time, this process may lead to widespread gene silencing in diverse dividing and nondividing cell types contributing to aging of the organism. PMID- 8231987 TI - Louis Bolk revisited. II--Retardation, hypermorphosis and body proportions of humans. AB - A number of morphological particularities of humans, generally thought to be specializations produced by natural selection, arguably are instances of hypermorphosis: alterations in proportion brought about by simple prolongation of general embryonic allometries. The descended larynx, the flattened human chest, and several limb proportions are given as examples. The case of the human foot, often quoted as an example of advanced specialization in humans, is examined in some more detail. Besides, it is argued that the human growth pattern displays a unique 7-year periodicity unlikely to be the result of the erratics of Darwinian evolution. Developmental acceleration is the unavoidable correlate of animal specialization; it brings about the breakdown of the periodicity present in the generalized humanlike development and causes the embryonic allometries to be cut off prematurely in the non-human mammal. PMID- 8231988 TI - Syphilis as AIDS?--A call for research. AB - Based upon a literature survey, this paper outlines the argument that Treponema pallidum may be the etiologic agent of AIDS. AIDS and syphilis have similar epidemiology, immunology and pathologies. In addition, the current methods of detecting and treating syphilis are inadequate. The similarities between syphilis and AIDS support the need for further research into their relationships, including the role Treponema pallidum may play in AIDS etiology. PMID- 8231989 TI - Oscillators in the human body and circular-muscle gymnastics. AB - There is a growing body of literature about the role of oscillators in the living body, and about the interactions between different oscillators. Considering the importance of endogenous oscillators in regulating the body's functions, and the existence of 'dynamical diseases', diseases of control systems which involve oscillators in the body, a way to mend dysfunctioning oscillators seems to be needed. Circular-muscle gymnastics, a method of physical activity which has been developed in Israel, reveals some phenomena which may point in a promising direction. Some of these phenomena call to mind known facts and theories about oscillators and their effects. PMID- 8231990 TI - Using neural networks to model personality development. AB - A neural network approach to modeling the development of personality traits through social learning is presented. From the more general model the special case of a network mapping four situation dimensions (input neurons) into seven dimensional personality traits (output neurons) is described. This network is allowed to learn with input/output sets representing conditions suspected of leading to a borderline personality disorder. The network's ability to learn these pattern pairs is demonstrated. The trained network is then presented with new input (situational) patterns and is shown to respond to these new situations with output patterns consistent with a borderline personality disorder. The neural network model is thus shown to have important advantages over other personality models in that it can predict what situations will produce shifts in personality traits, for example from active to passive. This model provides a quantitative and reproducible framework within which to discover and test theories of personality development. It is hoped that it will extend our ability to predict human behavior. PMID- 8231991 TI - Possible reversal of ageing and other mitochondrial deficiencies through retroviral transfection of mitochondrially encoded proteins to the nucleus. AB - This paper puts forward the hypothesis that ageing processes and a number of mitochondrially related diseases can be reversed or cured respectively. It briefly discusses the role of the mitochondrial genome in ageing and a number of inherited and age-related diseases. A rationale is then suggested which involves the utilization of retroviruses in translocating mitochondrial encoded proteins to the nucleus. Finally a number of problems which may arise from this translocation are discussed. PMID- 8231992 TI - On a possible explanation as to why most people have small refractive errors. AB - The case is made that there must be a feedback mechanism operating during the development of the human eye which means that over a population most people have relatively small refractive errors. The possible feedback mechanisms are examined and the conclusion is drawn that chromatic aberration of the human eye may produce visual cues which could be utilized in such a feedback system. PMID- 8231993 TI - Salt restriction in hypertension. AB - Salt has had a long and controversial history. It is not known when man began to use salt in his diet, but it is logical to believe that salt has always been a part of human diet though we do not have scientific proof for this. Restriction of salt in hypertensive patients has been a popular measure adopted world wide. This paper examines the usefulness of salt restrictions and the effect it would cause. Most cases of hypertension are symptomless--using a therapy which drastically affects a patients food habits is a poor strategy to ensure compliance. We conclude that salt restriction is not very useful and probably harmful in some patients. Additionally it reduces the quality of life. PMID- 8231994 TI - REM sleep pathways and anticholinesterase intoxication: a mechanism for nerve agent-induced, central respiratory failure. AB - The mechanism of death following exposure to anticholinesterases, such as the highly toxic nerve agents soman and VX, and other organophosphate anticholinesterases such as the insecticide parathion, remains unclear, although evidence from nerve agent research suggests that death occurs by an atropine blockable respiratory failure mediated through mechanisms involving the central nervous system. It is proposed that REM sleep pathways, which can be triggered by acetylcholine accumulation in the pontomedullar reticular field, mediate respiratory failure through the inhibition of respiratory muscles. Cholinergic activation of REM sleep activities may also account for other physiological and behavioural effects that follow exposure to nerve agents. These include forebrain activation, which is associated with EEG desynchronization and seizures, locomotor depression with concomitant loss of righting reflex, and limb jerks and extensions. Pharmacologic evidence for atropine and clonidine protection against soman intoxication effects is entirely consistent with a scenario of cholinergic receptor activation in the pontomedullar reticular field. PMID- 8231995 TI - Selenium--its biological perspectives. AB - Selenium is an essential trace element at lower concentrations and toxic at higher concentration. Animals can metabolize both inorganic and organic forms and convert non methylated Se to mono--or di--or tri--methylated forms, of which, mono-methylated forms are most toxic. Glutathione reductase converts selenoglutathione to H2S in liver and erythrocytes and is ultimately excreted. Se effects the toxicities of xenobiotic agents, provides antagonistic effect to Sulphur and co-administration with Zn increase Se retention in certain organs. At its toxic level (4-8 ppm) it increases Cu contents of heart, liver and kidney and has detoxifying or protecting effect against Cd and Hg. It is a prosthetic group of several seleno metalloenzymes. The concentration of the element is decreased in serum/plasma or erythrocytes of patients of AIDS, trisomy-21, Crohn's and Down's syndrome, phenylketonurea, Keshan's disease and cancer. Rather, the element has antiproliferative and cancer protecting effect. Se content of testes increases considerably during pubertal maturation and, during Se deficiency, the supply to the testes has priority over the other tissues. The element is localized in the mitochondrial capsule protein (MCP) and is involved in biosynthesis of testosterone. Neither the age of mother nor the concentration of Se during pregnancy has any effect on weight of baby or the length of pregnancy. Se levels in human milk is affected by maternal intake and its requirements by infants and young children are higher for their rapid growth. Clinical symptoms of its toxicity include severe irritations of respiratory system, metallic taste in mouth, formication of nose, signs of rhinitis, lung edema and brancho pneumonia. The typical garlic odour of breath and sweat is due to dimethyl selenide. PMID- 8231996 TI - The possible role of intravenous immunoglobulin in preventing pre-eclampsia. AB - Our understanding of the pathophysiologic abnormalities contributing to pre eclampsia has increased substantially over the past decade. This has been accompanied by the introduction of various methods which reduce the incidence of pre-eclampsia to approximately the same range (4-11.8% in high risk group), by attempting to correct the pathophysiologic abnormalities involved, rather than averting its occurrence. Numerous reports suggest that pre-eclampsia is caused by an abnormal maternal immune response to antigenic challenge by the fetoplacental allograft. If this assumption is true, it may be possible to manipulate the maternal immune response to fetal allograft and to assist the immunologic homeostasis of the maternal host with her conceptus by passive immunization with intravenous immunoglobulins (IVIg) and thus, to prevent pre-eclampsia. Recently IV preparations of Ig have become available for clinical use including treatment of various immunologic disorders during pregnancy. The effectiveness of this new mode of therapy can be related to several immunological mechanisms such as blockade of antibody binding to receptors on macrophages, increase in T suppressor cells, or decrease in antibody synthesis. The latter effect may be mediated by anti-idiotypic antibodies in the Ig preparation, which may bind to idiotypes on pathogenic autoantibodies and neutralise their activity. PMID- 8231997 TI - The membrane cluster hypothesis of mitogenesis and carcinogenesis. AB - This paper modifies and extends an earlier one on the same subject. It explains why external (but not internal) surface molecules of plasma membrane clusters may be rapidly scattered by any external challenging bioelectrical field. Temporary clusters from challenges may induce mitosis in cells near wounds and in epithelial stem cells. Weak challenges of much longer duration may initiate carcinogenesis by permanent clusters. Basic intracellular ligand/receptors or oncogene products in sufficient concentration at the membrane inner lipid layer may form permanent clusters rapidly. Additive increase of inner surface clusters by initiating agents is equated to promotion; accelerated cluster growth to progression. As a malignant cell grows, its cluster population increases until its membrane becomes permeable enough to stimulate mitosis. A progression mechanism is suggested that is consistent with the known properties of ras p21 proteins. The effect of long term exposure to power transmission line fields on mitosis and carcinogenesis is discussed. An approach to anticancer therapy is suggested, using a hypothesis-based mechanism for the anti-cancer activity of retinoic acid. PMID- 8231998 TI - The stomach as a system and the pathogenesis of experimental ulcer. AB - The stomach is prone to ulceration because of the hostile environment that exists within its lumen. The most important etiologic factor remains a topic of debate. We have considered the stomach as a system to lend insight into which pathophysiologic mechanisms might be most important. Systems are described by their content, hierarchy, entropy and interactions. The states of health, disease and death (i.e. ulceration) are represented by progressively increasing levels of entropy. It is argued that many of the purported causes of ulcer disease, such as acid back-diffusion or alcohol related necrosis, represent alterations in the system that affect small groups of cells that are low in the hierarchy and cause the tissue to enter the diseased state. We hypothesize that because blood flow is high within the hierarchy it represents the major homeostatic mechanism. If blood flow responds appropriately the system may return to the healthy state. If it does not the death of the system results in ulceration. Experimental evidence to support these contentions is presented. PMID- 8231999 TI - Primary Na+/H+ exchanger dysfunction: a possible explanation for insulin resistance syndrome. AB - Insulin resistance has been recently distinguished as a syndrome associated with a clustering of metabolic disorders, including non-insulin dependent diabetes mellitus (NIDDM), obesity, hypertension, dyslipidemia and atherosclerosis. To date, it is thought that all of these disorders are the resulting consequences of primary insulin resistance. We propose that insulin resistance and the metabolic diseases mentioned can be caused by primary overactivity of the Na+/H+ exchange. This hypothesis has practical connotations for understanding the pathogenesis of the insulin resistance syndrome. PMID- 8232000 TI - The role of cartilage-derived antigens, pro-coagulant activity and fibrinolysis in the pathogenesis of osteoarthritis. AB - Superficial fraying, splitting and fibrillation of articular cartilage as a consequence of ageing or mechanical injury is not always associated with joint pain. However, similar cartilage lesions accompanied by synovitis and engorgement of the subchondral vasculature generally is; the disorder being known as osteoarthritis (OA). In this hypothesis it is contended that the progression of early cartilage fibrillation to symptomatic OA arises as a consequence of the antigenic nature of cartilage components which when released into synovial fluid and the circulation can stimulate leukocytes (to a varying degree) to produce a number of factors some of which promote blood coagulation. While it is known that with ageing hyper-coagulation and plaque deposition is increased, the augmentation of this process by factors released by activated leukocytes is considered to exacerbate the problem. These haematological events may be particularly relevant in individuals whose leukocytes are hypersensitive to the cartilage-derived antigens, and whose fibrinolytic system is less capable of mobilizing the thrombi deposited in synovial tissues and the subchondral vascular tree. PMID- 8232001 TI - On the causes of multiple sclerosis. AB - Evidence on aetiology in multiple sclerosis suggests that the prevalence depends on the interaction of two factors, diet and exposure to visible sunlight. The dietary features which may be beneficial include supplementation with fish oils, avoidance of saturated fats, and the associated intake of antioxidants with unsaturated fatty acids. Inhibition, by antioxidants, of the enzyme lipoxygenase inhibits leukotriene synthesis, and the presence of fish oils leads to the production of leukotrienes with less inflammatory properties. This is of particular importance in the retina where leukotrienes might be the underlying cause of retrobulbar neuritis. The antioxidant properties of vitamin A may also lead to inhibition of leukotriene synthesis. Visible solar radiation could be of benefit therefore by releasing vitamin A from visual pigment rhodopsin. The interaction of these two factors may explain the epidemiological observations on the prevalence of multiple sclerosis. PMID- 8232002 TI - Granted publications: taken for granted? AB - Highly respected journals now relatively rarely publish independent research which is not grant-supported. This may lead to a reduction in the publication of innovative work. PMID- 8232003 TI - Illicit amphetamine use as a public health problem in Australia. PMID- 8232004 TI - Is cholesterol testing/treatment really beneficial? PMID- 8232005 TI - Digitalis for patients with heart failure in sinus rhythm. PMID- 8232006 TI - Aspirin and non-steroidal anti-inflammatory drugs in the chemoprevention of colorectal cancer. PMID- 8232007 TI - Neck strain in car occupants. The influence of crash-related factors on initial severity. AB - OBJECTIVE: To examine the relationship between the initial severity of neck strain in car occupants and crash-related factors, in particular, crash severity. DESIGN/PARTICIPANTS: Thirty-two individuals with neck strain after a car crash, drawn from physiotherapy and general practices in metropolitan Adelaide, were interviewed about their experience and examined by a manipulative physiotherapist. Crash severity was assessed by measurement of damage to the involved vehicles. MAIN OUTCOME MEASURES: Five measures of neck strain severity were obtained: number of body regions with symptoms, number of positive responses to palpation, cervical range of motion, subject's own rating on an analogue pain scale, and examiner's severity rating. Two measures of crash severity were used: maximum residual deformation, and velocity change of the subject's vehicle. RESULTS: Neck strain was observed in individuals who were involved in crashes of low severity. Maximum residual deformation of the vehicle was negatively associated with the subject's cervical range of motion and positively associated with the other measures of neck strain severity. For rear impacts, both measures of crash severity were associated with measures of neck strain severity. The group of subjects who were aware of the impending collision had a greater range of cervical spine movement and fewer positive responses to palpation than those who were unaware. CONCLUSIONS: Initial severity of neck strain is positively correlated with crash severity. Awareness of the impending collision may have a mitigating effect on injury severity. PMID- 8232008 TI - Cervical cancer screening in Western Australia in 1992: progress since 1983. AB - OBJECTIVES: To estimate the rate of cervical cancer screening in Western Australia in 1992, and any variation by age, place of residence, and socio economic status; and to determine the proportion of smears taken by different service providers. DESIGN: Descriptive study; collection of data from Papanicolaou (Pap) smear request forms during one calendar month. SETTING: All 13 cytology laboratories in Western Australia. PARTICIPANTS: 15,767 women in Western Australia aged 15 years and over having a cervical smear in March 1992. MAIN OUTCOME MEASURES: Rates by age of cervical cancer screening per 1000 woman-years; age-standardised rate ratios for socioeconomic status and place of residence; proportion of smears taken by male and female service providers. RESULTS: The estimated rate of Pap smears at ages 15 years and over was 303 smears per 1000 woman-years, an increase of 44% over a similar survey in 1983. The greatest increases were among women aged 50 years and over, but their rates were still well below that equivalent to three-yearly smears. Differences in the rate of screening by socioeconomic status (defined by residential postcode) were not statistically significant. The age-standardised rate ratio comparing country women with women in the Perth metropolitan area was 0.91 (95% confidence interval 0.87-0.94). General practitioners took 78% of the smears, and at least 46% of all smears were taken by female service providers. Almost all the increase in the screening rate since 1983 could be attributed to an increase in the rate of smears per 1000 woman-years taken by female general practitioners. CONCLUSION: While there have been marked improvements in the rates of cervical screening in Western Australia over the past nine years, there are still major deficiencies in the screening coverage of women aged 50 years and over. PMID- 8232009 TI - The investigation of chest pain: audit and intervention. AB - OBJECTIVE: To examine the patterns of tests requested for patients admitted with chest pain, and to monitor the effects on those patterns of issuing a set of guidelines formulated by an expert panel. SETTING: Tertiary referral hospital (teaching). DESIGN: A retrospective audit of the patterns of testing of patients with chest pain compared with testing recommended by an expert panel, followed by comparison with patterns of testing after guidelines were issued. INTERVENTION: Practice guidelines, based on the expert committee's recommendations, were drawn up for use in the cardiology unit. OUTCOME MEASURES: Tests per admission, bed days per admission, emergency readmission rate. PATIENTS: Sixty-seven patients with diagnosis at the time of admission of chest pain and a discharge diagnosis of "intermediate coronary syndrome". RESULTS: The major finding of the audit was the excessive use of "biochemistry test profiles" instead of targeted tests as recommended by the expert committee. Intervention led to a decrease in the number of tests per admission in the cardiology unit from an average of 22.9 tests in the six months before intervention to 9.7 in the six months after (P < 0.001). There was no effect on bed days per admission or emergency readmission rates. CONCLUSION: Audit and intervention in a specialised unit of patients with a specified diagnosis led to more cost effective use of the resources of the Royal Melbourne Hospital Department of Biochemistry. PMID- 8232010 TI - The epidemiology of tuberculosis in Victoria. AB - AIMS: To describe the epidemiology of Mycobacterium tuberculosis (MTb) in Victoria, Australia. METHODS: We analysed data from the Victorian Mycobacterium Reference Laboratory database and MTb notifications to Health and Community Services Victoria, for the period 1948 to 1992. RESULTS: The annual number of notified tuberculosis cases peaked at 1143 in 1954 and declined to 266 in 1992. The incidence rate decreased from 47 to 6.2 per 100,000 in the same period. The pattern of tuberculosis has changed significantly over the last two decades, reflecting the high prevalence of MTb in recent immigrants, particularly in those from South-East Asia. Among people born overseas the number of cases increased from 40% of notifications in 1970 to 80% in 1990; they are younger, more likely to be female and to have extrapulmonary disease, than their Australian-born counterparts. Between 1987 and 1991, the mean annual incidence of tuberculosis in people born in Australia and South-East Asia respectively was 1.5 and 47.5 per 100,000. People born in Vietnam, Cambodia and the Philippines had rates between 100 and 400 per 100,000. These rates have remained stable during the last six years. Almost 50% of migrant patients presented within five years, and 30% within two years, of arrival in Australia. The estimated mean annual incidence of MTb in homeless men between 1984 and June 1992 was 80 per 100,000. Resistance to all first-line drugs increased from 10.8% of all isolates in 1981 to 14.7% in 1990. The incidence of multi-drug resistant MTb (MDR-MTb) has been consistently less than 2% of isolates per year in the past 15 years. There were 14 cases of MTb and HIV between 1985 and June 1992. The mean annual incidence of MTb in people with AIDS is 1579 per 100,000 (range, 570-2420 per 100,000), with a relative risk of 236 (95% confidence interval [CI], 134-414). MDR-MTb has not been documented in HIV-infected individuals in Victoria. CONCLUSION: Tuberculosis remains an important public health concern. Groups at high risk include people born overseas, the homeless, and people infected with HIV. Physicians caring for AIDS patients should consider the diagnosis of MTb, since its incidence in AIDS patients is so high. Our data suggest possible shortcomings in current methods of screening and prophylactic treatment of migrants and refugees. The rising rate of drug resistance cannot be ignored. We should consider strategies to ensure judicious use of anti-tuberculous drugs by physicians, and optimal patient compliance. These issues are critical to the future of tuberculosis control in Victoria. PMID- 8232011 TI - Pharmacological considerations in digoxin therapy. PMID- 8232012 TI - Prevention of health problems in later life. AB - The areas of health promotion and disease prevention for older people have been neglected in medical practice. Many doctors, as well as the general community, believe that for people over 65 years of age it is too late for preventive care. This assumption is incorrect. However, there are a number of issues that are unique to prevention of health problems in later life. The goals of preventive care are broader than the issues of illness and life expectancy. Quality of life, autonomy and maintaining optimal function are equally important concerns for older people. Practising prevention involves not only reducing primary risk factors or screening for disease, but also encompasses psychological, environmental and social issues as these affect health and well-being. In addition, special medical problems require consideration; these include falls, incontinence, confusion, poor mobility and iatrogenic disease which are not the result of one disease process, but have many inter-related causes. Greater prominence should be given to patient education and counselling on these medical, social and psychological issues. General practitioners are ideally placed to initiate prevention as 87% of older Australians visit their GP once a year. While clear recommendations can be given on the effectiveness of some interventions, for others the doctor involved will have to make an informed decision based on individual clinical circumstances. Preventive care in later life is an integral part of good geriatric medicine. PMID- 8232013 TI - Hepatitis in the tropics. AB - Viral hepatitis, caused by one of five different viruses, is an important cause of illness in tropical countries and a significant cause of death. Vaccines against hepatitis A and B are now available and, if used widely, have the potential virtually to eliminate both these diseases (and also hepatitis D). Vaccines against hepatitis C and E are being developed. PMID- 8232014 TI - Seeking the perfect match. HLA and transplantation. PMID- 8232016 TI - Valley of the shadow of misinformation. PMID- 8232015 TI - General practice. PMID- 8232017 TI - What are we eating? PMID- 8232018 TI - Thermography update. PMID- 8232019 TI - Vaccination against influenza infection. PMID- 8232020 TI - Reducing length of stay after hip fracture. PMID- 8232021 TI - A case report of Boerhaave's syndrome following colonoscopy preparation. PMID- 8232022 TI - Hypertension and diabetes in older people. PMID- 8232023 TI - Comparison of South Australian city and country general practitioners as regards reported asthma management. PMID- 8232024 TI - Lead encephalopathy and hallucinations. PMID- 8232025 TI - Screening for gestational diabetes. PMID- 8232026 TI - Aboriginal maternal mortality: whose problem? PMID- 8232027 TI - Deaths in custody. PMID- 8232028 TI - Clusters of birth defects. PMID- 8232029 TI - Basic pharmacology. PMID- 8232030 TI - Australian deaths in custody, 1980-1989. 1. Relative risks of aborigines and non aborigines. AB - OBJECTIVE: To assess the risks of death in police or prison custody in Australia for Aborigines and non-Aborigines. DESIGN, SETTING AND PARTICIPANTS: Retrospective collection and analysis of data about deaths occurring in police or prison custody in Australia in the 10-year period 1980-1989. RESULTS: For the period 1980-1989, Aboriginal and non-Aboriginal people in prison had risks of death similar to those of the total Australian population, but the risks of death in police custody for both Aborigines and non-Aborigines were between 10 and 20 times the risks of death experienced by the general population. Within each type of custody, there were no significant differences in the risks of death experienced by Aboriginal and non-Aboriginal people. CONCLUSION: Although some deaths in custody are inevitable, the very much higher risks of death in police custody for both Aborigines and non-Aborigines has implications for custodial authorities in terms of preventive and remedial action. Reduction of the excess of Aboriginal over non-Aboriginal deaths in police and prison custody relative to general population figures requires action to reduce the over-representation of Aboriginal people in custody and to improve the general well-being and health status of Aboriginal people in the community. PMID- 8232031 TI - Australian deaths in custody, 1980-1989. 2. Causes. AB - OBJECTIVE: To assess the causes of death for people in police or prison custody in Australia, with particular attention to the differences between Aboriginal and non-Aboriginal deaths. DESIGN, SETTING AND PARTICIPANTS: Retrospective collection and analysis of data about deaths occurring in police or prison custody in Australia in the 10-year period 1980-1989. RESULTS: In the period 1980-1989, 527 deaths are known to have occurred in police or prison custody. About half these deaths were due to self-harmful behaviour, including hanging. The next most frequent causes of death were diseases of the circulatory system, injuries and diseases of the respiratory system. CONCLUSION: The high number of deaths in custody resulting from self-harmful behaviour has important implications for the criminal justice and corrections systems. Minimisation of the number of people held in police and prison custody is important in preventing such deaths. Close attention needs to be paid to ensuring the safety of lock-ups and prisons and to the screening of people likely to be at risk of death from self-harmful behaviour. Custodial authorities have a clear responsibility to provide quality preventive and clinical health services to all people in custody. Special attention needs to be directed to the specific health needs of Aborigines, many of whom are at much greater health risk than are non-Aborigines. PMID- 8232032 TI - Birth size of Australian aboriginal babies. AB - OBJECTIVES: (i) To describe birth size of Aboriginal babies by sex, gestational age, and Aboriginality; (ii) to analyse the results with reference to standards of ponderal index and birthweight for gestational age. SUBJECTS: 570 liveborn singletons routinely delivered at Royal Darwin Hospital between January 1987 and March 1991, and recorded in the Delivery Suite Register as being born to an Aboriginal mother. MAIN OUTCOME MEASURES: Weight, length and head circumference at birth. RESULTS: The mean birthweight was 3098 g (standard deviation, 601 g), peak gestational age was 39 weeks, 13% were low birthweight and 7% were preterm. Preterm rates did not differ significantly for sex and Aboriginality. Babies without a non-Aboriginal ancestor had a lower mean birthweight and at term, were significantly smaller than babies with a non-Aboriginal ancestor as assessed by mean birthweight, length, head circumference and ponderal index. More than a quarter of babies (27%) without a non-Aboriginal ancestor were below the 10th percentile of birthweight for gestational age, compared with 14.2% of babies with a non-Aboriginal ancestor. CONCLUSIONS: On the basis of postnatal clinical estimates of gestational age, Aboriginal babies have a preterm rate of 7% and Aboriginal babies without a non-Aboriginal ancestor are smaller in size at birth than babies with a non-Aboriginal ancestor. PMID- 8232033 TI - Pseudomonas aeruginosa bacteraemia. Is pancreatobiliary disease a risk factor? AB - OBJECTIVE: To review changes in the epidemiology, course, and outcome of bacteraemia caused by Pseudomonas aeruginosa. DESIGN AND SETTING: A retrospective, descriptive study of consecutive cases of P. aeruginosa bacteraemia occurring at a university teaching hospital. PATIENTS AND METHODS: Between January 1980 and December 1989, 164 patients were admitted to Westmead Hospital with P. aeruginosa bacteraemia. Patients in whom there was no clinical evidence of sepsis were excluded from analysis leaving a cohort of 152 patients. Hospital records were reviewed and details of demography, clinical features, therapy, and outcome were recorded. RESULTS: One hundred and fifty-five episodes of P. aeruginosa bacteraemia were recorded at an average rate of 0.39 per 1000 admissions per year. The respiratory and pancreatobiliary tracts were the most common sources of the bacteraemia. Pancreatobiliary disease, independent of an underlying malignancy or immunosuppression, emerged as a previously undescribed risk factor for pseudomonal bacteraemia (incidence of 3.0 episodes per 1000 hospital admissions for patients with this disease). The crude mortality rate was 52%; 35% was attributable to pseudomonal bacteraemia. Factors identified as being independently associated with an increased mortality included hypotension, age of 60 years or older, and the presence of an underlying malignancy. Combination therapy with an antipseudomonal penicillin and an aminoglycoside confers a significant survival advantage independent of underlying neutropenia. CONCLUSIONS: Bacteraemia caused by P. aeruginosa remains an important cause of morbidity and mortality. Pancreatobiliary disease represents a new risk factor for P. aeruginosa bacteraemia, independent of an underlying malignancy or immunosuppression. It may be prudent to consider P. aeruginosa as a cause of sepsis in these circumstances, especially if there has been instrumentation of the biliary tree. Hypotension, age of 60 years or older, and the presence of an underlying malignancy were independently associated with significantly increased mortality. Appropriate antibiotic therapy consisting of an antipseudomonal beta lactam in addition to an aminoglycoside resulted in a significant decrease in mortality compared with the use of an aminoglycoside alone, not only in the study population as a whole, but also in patients without neutropenia. PMID- 8232034 TI - Typhoid in 1993. AB - Typhoid remains a disease of major importance world-wide although improvements in public health have made it an exotic disease in developed countries like Australia. Effective antibiotic therapy with the advent of chloramphenicol, which was first used to treat typhoid in the 1940s, has also dramatically altered the natural course of the disease and reduced its mortality rate from around 25% to as low as 1%. The main areas of recent change include the emergence of resistance to previously effective antibiotics, more aggressive intervention in the management of severe typhoid and some of its complications such as perforation, and the development of an oral typhoid vaccine that may replace the equally effective but more unpleasant parenteral vaccination that has been widely used since World War. PMID- 8232035 TI - Hypertension. PMID- 8232036 TI - Pharmacological agents used in the treatment of cardiovascular disorders. PMID- 8232037 TI - Innovative teaching techniques to enhance general practice training. AB - Two techniques to improve the quality of general practice training are described. These techniques are used during visits to trainee doctors in their routine general practice work. PMID- 8232038 TI - Family screening for genetic haemochromatosis. What is the most effective method of contact? AB - OBJECTIVE: To evaluate the effectiveness of various approaches to follow-up screening of family members after genetic haemochromatosis (GH) has been diagnosed in an individual. DESIGN AND SETTING: Thirty-eight patients diagnosed with GH at the Flinders Medical Centre and Repatriation General Hospital in South Australia over a 16-year period were identified by review of case notes. If possible, a questionnaire and a follow-up interview were used to obtain information about the screening that had been done and any obstacles to screening that were encountered. Patients were asked to outline a family tree, and indicate who had and who had not been screened and reasons for not screening. EVALUATION CRITERION: The number of relatives screened or not screened. RESULTS: The families of 71% of patients diagnosed with haemochromatosis were subject to some degree of screening, although complete screening was achieved in only 13%. Approximately a third (30%) of our GH patients were detected by family screening. However, not all relatives who were at risk were screened. Reasons given for not screening were, for example, ignorance about "genetic disease" and fear of treatment. CONCLUSION: Family screening is an important means of detecting new cases of GH. If these findings reflect practice elsewhere, there is a need for education of medical practitioners regarding those who should be screened. More easily accessible information for patients with GH and their relatives would help to overcome some of the problems surrounding identification of the disease. PMID- 8232039 TI - Adult advanced life support. The Australian Resuscitation Council Guidelines. PMID- 8232040 TI - Drug-induced neurological disorders. PMID- 8232041 TI - Gastroenterology. PMID- 8232042 TI - DT vaccine in place of DTP vaccine for children. PMID- 8232043 TI - The use of poorly validated expensive technology. PMID- 8232044 TI - Medical victims of pathological love--the hippocratic curse? PMID- 8232045 TI - Unusual clinical problem associated with obesity surgery. PMID- 8232046 TI - Medicine and surgery inside the fallopian tube. PMID- 8232047 TI - Obstetrics in crisis? PMID- 8232048 TI - Asthma "six point plans" and beyond. PMID- 8232049 TI - Glue: what's really in it for sniffers. PMID- 8232050 TI - Injecting drug use: a continuing and major vehicle for transmission of infection. PMID- 8232051 TI - Strongyloidiasis in the Northern Territory. PMID- 8232052 TI - The likelihood of pregnancy with IVF and GIFT in Australia and New Zealand. PMID- 8232053 TI - Initial Australian experience with the atypical antipsychotic clozapine. PMID- 8232054 TI - Correction of error: Methadone dosage and retention of patients in maintenance treatment. PMID- 8232055 TI - Tick disease warnings. PMID- 8232056 TI - A comparison of country and metropolitan general practice. AB - OBJECTIVE: To provide a description of country general practice in Australia, and to determine the extent to which country and metropolitan general practice differ in terms of the characteristics of the practitioners, the morbidity managed, treatments provided and the availability of support services. METHOD: A survey requiring the recording of details of all direct and indirect patient encounters on encounter forms by a sample of general practitioners (GPs). Each GP recorded for two one-week periods separated by an interval of six months, between October 1990 and October 1991. The recording weeks were spread as evenly as possible throughout the year. SAMPLE: Participants were drawn from a list of medical practitioners in Queensland, NSW and Victoria who provided more than 1500 general practice Medicare items of service during the previous year. The sample was stratified within States by population of postcode, into metropolitan areas and three country strata: "small country towns" (population less than 5000); "medium country towns" (5000-15,000); and "large country towns" (more than 15,000). The total country sample is referred to as "country areas". Planned sample size was 180 country GPs (20 in each stratum in each State) and 60 metropolitan GPs (20 in each of the three State capital cities). The final data set was weighted to be representative of the distribution of the source population. DATA COLLECTION: The variables studied included: GP characteristics; practice isolation factors; patient age, sex and status to the practice; patient reasons for encounter (up to three per encounter); problems managed (up to four); drugs prescribed and other treatments provided (up to four per problem); tests and investigations ordered and referrals made at these encounters; and planned follow-up. Data were centrally coded. Participation rate: The final sample of 231 GPs (177 country and 54 metropolitan) recorded information during 435 recording weeks (336 country and 99 metropolitan). These practitioners represented 50.7% of those contacted and available, the response rate being better in country (57.5%) than in metropolitan (36.5%) areas. A practice profile questionnaire which included questions regarding the doctor and the practice was completed by 97.4% of participants, while a questionnaire on distance from support services was completed by 93.8% of country participants. The final weighted data set included 51,741 encounters with country GPs and 11,351 with metropolitan GPs. RESULTS: The general practitioners: Country GPs were less likely to be female or to conduct consultations in a language other than English, and were more likely to do some work on a salaried or sessional basis. GPs from small country towns were older, more likely to be in solo practice, and more likely to belong to a professional organisation. "Remoteness" of towns: Nearly all towns were within 25 km of a hospital, but far fewer small and medium country towns were within 50 km of a base hospital than large country towns. X-ray services were almost universally available within 50 km, and with the exception of small country towns so were pathology services. Access to medical specialists and to a lesser degree other health professionals decreased with population size--patients in 30% of small towns had to travel over 100 km to see many specialists and some health professionals. Self-reported procedural work: GPs in small and medium country towns were more likely than those in large towns to report performing procedural work, the largest difference being in the area of em PMID- 8232057 TI - [Pathophysiology of kidney diseases]. PMID- 8232058 TI - [Urinary tract infections]. PMID- 8232059 TI - [Drug therapy of urinary incontinence in women]. PMID- 8232060 TI - [Psoriasis--pathogenesis and treatment]. PMID- 8232061 TI - Surgical treatment of Parkinson's disease. PMID- 8232062 TI - Tetramune--a combined vaccine for infants. PMID- 8232063 TI - Butorphanol nasal spray for pain. PMID- 8232064 TI - Felbamate. PMID- 8232065 TI - Rimantadine for prevention and treatment of influenza. PMID- 8232066 TI - Cationic glycoproteins in sera of patients with acute infections identified as kappa light chain glycosylated IgG. AB - In half of the sera from patients with acute bacterial infections and 15% of the sera from patients with acute viral infections glycoproteins were found that form a scalariform pattern in the cationic range upon isoelectric focusing. The cationic glycoproteins appeared with the clinical illness. After subsidence of the symptoms they disappeared within 4 to 6 weeks. The proteins were identified as immunoglobulin G (IgG) by determination of the NH2-terminal amino acid sequence. Remarkably, these IgG only contained light chains of the kappa type with high proportions of carbohydrates. Both, N-glycosidic- and O-glycosidic bound glycans were present. The glycosylated light chains may render the cationic IgG multireactive. Thus, it may be part of an early nonspecific immune defense mechanism. PMID- 8232067 TI - Polymerase chain reaction and direct sequencing of Neisseria gonorrhoeae protein IB gene: partial nucleotide and amino acid sequence analysis of strains S4, S11, S48 (serovar IB4) and S34 (serovar IB5). AB - A pair of primers were designed for the polymerase chain reaction (PCR) to amplify a 341-base pair fragment of the gene encoding the outer membrane protein IB (PIB) of Neisseria gonorrhoeae. This PCR technique is specific and sensitive, being able to detect gonococcal strains belonging to ten different PIB serovars, but not PIA gonococcus nor other negative control bacteria. PCR products of four representative PIB strains were directly sequenced. Of the three strains belonging to serovar IB4, two (S11 and S48) shared identical nucleotide and amino acid sequences in the PIB region examined. The third IB4 strain (S4) revealed sequences identical to the published IB26 strain (P9). The sequences of strains P9, S4, S11 and S48 were found to differ from those of strain S34 (serovar IB5). The PCR sequencing technique can further differentiate strains belonging to a common serovar and establish clonal relationships among strains. As a molecular epidemiological tool, the PCR-sequencing strategy can augment existing typing methods including serotyping. PMID- 8232068 TI - Sensitivity of HIV antibody detection in saliva. AB - To assess the sensitivity and specificity of HIV antibody detection in saliva we tested matched serum and saliva samples from HIV-infected and uninfected individuals. Saliva specimens were collected by two different devices of the Salivette system and stored at different temperatures. Samples were tested for HIV antibodies by two commercially available enzyme-linked immunosorbent assays (ELISAs; Wellcome, Biotest). HIV antibodies were detected in 98.5% (Wellcome) and 97.8% (Biotest) of the saliva samples (standard Salivettes) from 135 seropositive individuals. Using the Salivettes flavoured with citric acid the sensitivity was only 22.9%. No reactions in ELISA were found in saliva from HIV-seronegative individuals. Salivary HIV-specific IgA was detected in 90% of seropositive individuals. All positive saliva samples stored at room temperature were still reactive after 20 days; of those stored at 37 degrees C, 23 out of 24 were positive when retested on day 5. Sensitivity of HIV antibody detection in saliva samples dried onto filter paper was 100% when a minimum of 100 microliters of saliva was used. HIV antibody testing in saliva is an efficient tool for large scale epidemiological studies when standard Salivettes are used for sample collection. Saliva samples can be stored in Salivettes or dried onto filter paper for several days at room temperature and under tropical conditions (37 degrees C). PMID- 8232069 TI - A guide to the use of pore-forming toxins for controlled permeabilization of cell membranes. AB - Depending on the size of the pores one wishes to produce in plasma membranes, the choice will probably fall on one of the three toxins discussed above. S. aureus alpha-toxin should be tried first when pores of 1-1.5 nm diameter are required. This is generally the case when Ca2+ and nucleotide dependence of a given process is being studied. If alpha-toxin does not work, this is probably due to the fact that the toxin either does not produce pores, or that the pores are too small. In this case, high concentrations of alpha-toxin should be tried. If this still does not work, we recommend the use of HlyA. When very large pores are to be created, e.g. for introduction of antibodies into the cells, SLO or another member of this toxin family are the agents of choice. SLO preparations need to be checked for presence of protease contaminants. Tetanolysin currently offers advantages since it is protease-free, and the size of the pores can probably be controlled by varying the toxin dose. Methods for assessing the size of pores created by such agents have been published in the recent literature, and the appropriate papers can be consulted whenever the need arises. PMID- 8232070 TI - 2nd International Workshop on Pore-Forming Toxins. September 29-October 2, 1993, Mainz, Germany. Abstracts. PMID- 8232071 TI - Health care in Africa--which way? AB - This article gives a personal view of the health situation in a typical country in the South. Uganda is a country which is well endowed with natural resources, as are many countries in the South, but is plagued by poverty, conflict, endemic infectious diseases and, more recently, disease due to HIV. The article argues for an integrated solution to appropriate health care, proposing that good health results from sustainable development. It focuses on constraints to development such as conflict, militarization, environmental degradation, lack of community action, and inappropriate policies by both developing world governments and members of the international arena. All of these play a role in the achievement of sustainable equity world-wide. PMID- 8232072 TI - The human dimensions of international debt. AB - The burden of international debt in Latin America and Africa has caused serious health damage during the 1980s. It has been responsible for the deaths of many thousands of young children in the developing world. Despite efforts to solve the problem, debt will probably persist into the next century, draining the life blood of poor countries. Reasons are discussed as to why doctors should take an active part in opposing the policies of the major financial institutions responsible for the reverse flow of capital from South to North. PMID- 8232073 TI - Second malignant tumors following treatment during childhood and adolescence for cancer. AB - Many pediatric and adolescent cancer patients are treated with carcinogenic chemotherapeutic agents and radiation therapy to achieve permanent control of their malignancy. These modalities may induce a new cancer in the successfully treated patient. To identify disease and treatment factors which increased the risk of occurrence of a second malignant tumor following modern treatment for cancer during childhood or adolescence, we reviewed the courses of 1,406 previously untreated patients who were less than 20 years of age at diagnosis and were treated at Roswell Park Cancer Institute between January 1, 1960 and December 31, 1989. Eighteen patients developed a second malignant tumor, including two meningiomas, 2.65-25.65 years after diagnosis of the first cancer. The actuarial risk of a second malignant tumor was 5.6% at 25 years after diagnosis. Using Cox proportional hazards modelling, we identified prior therapy with BCNU (P = 0.0055) and doxorubicin (P = 0.0254) as the only factors that were significantly associated with the risk of a second malignant tumor. Three second malignant tumors of the central nervous system occurred following treatment with a nitrosourea. Successfully treated patients must be carefully followed to identify treatment related malignant tumors at an early stage. PMID- 8232074 TI - Autologous bone marrow transplantation for pediatric Wilms' tumor: the experience of the European Bone Marrow Transplantation Solid Tumor Registry. AB - This survey includes 25 children with Wilms' tumor undergoing high-dose chemotherapy associated with autologous bone marrow transplantation (ABMT) in the period June 1984-December 1991 and enrolled in the European Bone Marrow Transplantation Registry for Solid Tumors. At diagnosis, 12 children presented stage IV disease, 5 stage III, 3 stage II, and 5 stage I. Before ABMT, 21 children had 1 to 4 relapses (median 1); 13 achieved a second or subsequent complete remission (CR), four stage IV children failed to respond to first line treatment and achieved either CR (3 patients), or partial remission (PR) after second line therapy. At high-dose chemotherapy, 17 children were in CR and 8 had measurable disease. Seven different high-dose regimens were administered, even if 20 children received melphalan mostly associated with vincristine and 8 involved field radiotherapy. Three children died early of pneumonitis; 2 developed an acute transient renal failure, 1 a chronic renal failure. Out of the 8 children with target disease at graft, 2 died of toxicity, 5 achieved CR, 1 obtained PR, and only 1 is presently alive in CCR at 39 months after ABMT. Of the 17 children grafted in CR, 8 are alive event-free at 14-90 months (median 34) from ABMT; 7 relapsed at 3-23 months (median 7 months); 1 died of toxicity and 1 was lost to follow-up in CR at 12 months. A salvage attempt with high-dose chemotherapy in children with resistant or poor prognosis recurrent Wilms' tumor seems to be justified. An international cooperative protocol taking into account the increased risk of lung and renal toxicity is necessary. PMID- 8232075 TI - Bone and gallium scintigraphy in children with rhabdomyosarcoma: a 10-year review. AB - Technetium-99m bone and Gallium-67 scintigraphy has been widely used in the management of children with solid tumors. A retrospective review of all patients with rhabdomyosarcoma (RMS) from January 1980 to December 1989 was undertaken in order to determine the sensitivity and specificity of scintigraphy in detection of metastatic disease and to determine optimum frequency of serial scans in follow up. Over the 10-year period, 40 patients were diagnosed and treated for RMS: 22 were newly diagnosed from 1980-1984 and of these 16 had a minimum 5-year disease-free period whilst 6 died of the disease, giving a cure rate of 73%. A total of 271 bone and 236 Gallium-67 scans were performed. With respect to detection of metastatic disease in all tissues, Gallium-67 scans had a sensitivity of 84%, specificity 95% and bone scans had a sensitivity of 70% and specificity 95%. Bone scan sensitivity and specificity for skeletal metastases were 100% and 95%, respectively. Considering only patients with Gallium avid primary tumors, the Gallium-67 scan sensitivity was 94%. Ten patients developed new metastatic disease or primary recurrence after starting therapy, 8/10 within 12 months of diagnosis. Frequent surveillance scanning in the treatment phase, 2 years post-diagnosis, is of value in the early detection of metastases and monitoring disease response to therapy. Surveillance scanning after completion of treatment is more difficult to justify. PMID- 8232076 TI - Value of postchemotherapy bioptical verification of complete clinical remission in previously incompletely resected (stage I and II pT3) malignant mesenchymal tumors in children: International Society of Pediatric Oncology 1984 Malignant Mesenchymal Tumors Study. AB - Bioptical verification of complete clinical remission achieved with postoperative chemotherapy in 140 pediatric patients with nonradically resected stages I and II nonparameningeal malignant mesenchymal tumors was evaluated. Tumor histology was rhabdomyosarcoma in 92 and nonrhabdomyosarcoma in 48 patients. All were included in the International Society of Pediatric Oncology "1984 Malignant Mesenchymal Tumors Study." Treatment given after nonradical surgery and before the biopsy consisted of 3 to 6 IVA courses (ifosfamide, vincristine, actinomycin D). Complete clinical remission (no tumor at physical examination, X-rays, sonography, computed tomography, scintigraphy) was bioptically confirmed in 52 rhabdomyosarcoma and 23 nonrhabdomyosarcoma cases, whereas in all the remaining patients (39 rhabdomyosarcoma and 22 nonrhabdomyosarcoma) complete clinical remission was not bioptically verified. Forty-eight-month local relapse rates were 51% in bioptically confirmed and 48% in nonbiopsied rhabdomyosarcoma patients. In nonrhabdomyosarcoma cases, relapse rates were 25% and 14%, respectively. In 4 cases, bioptical specimens contained tumor cells. In spite of aggressive treatment, only 1 is actually in remission. The total 48-month relapse rate (biopsied and nonbiopsied) was significantly higher in rhabdomyosarcoma patients (50%) than in nonrhabdomyosarcoma patients (26%). The value of bioptical verification of complete clinical remission seems to be limited in our series, when known that relapse rate in bioptically confirmed complete remission cases remained high and the outcome in rare "positive biopsy patients" was poor. PMID- 8232077 TI - Ifosfamide-induced subclinical nephrotoxicity and its potentiation by cisplatinum. AB - Renal function was assessed in 72 children and adolescents 3.5 to 123 months after completion of chemotherapy employing ifosfamide (n = 39) or ifosfamide plus cisplatinum (n = 33). No patient had preexisting renal parenchymal disease. Whereas reduction in glomerular filtration rate was present in six of 69 patients (8.7%), impairment of tubular transport for phosphate, glucose, and amino acids was more frequent: 32.8% of the patients showed reduction in phosphate reabsorption, and glucose and amino acid reabsorption was lowered in 16.4% and 55.0%, respectively. Elevated sodium excretion was found only occasionally, and there was no evidence of renal tubular acidosis. Proximal tubular damage is related to ifosfamide chemotherapy, but correlation between ifosfamide dose and phosphate reabsorption was not linear. The most severe depletion of phosphate reabsorption was seen in patients treated with both ifosfamide and cisplatinum. On reexamination of phosphate reabsorption after a median interval of 8 months, the majority of patients with initially reduced values showed further deterioration of this function. PMID- 8232078 TI - A study of children, fathered by men treated for testicular cancer, conceived before, during, and after chemotherapy. AB - One hundred fifty children of 113 fathers with testicular tumour treated from 1979 on the National Institute of Oncology, Budapest, were studied. Three groups were formed on the basis of the time of conception; 69 children were born before the illness of the fathers, 40 during the 12 pretreatment months, and 41 during or after combined chemotherapy. One hundred fifty control children underwent tonsillectomy/appendectomy, but were otherwise healthy. They were matched according to age, sex, and place of inhabitance with index children. Family anamnesis, perinatal, and gestational data were listed; thereafter, physical, laboratory, immunological, and, if required, radiological examinations were made. No difference was detectable in the somatic and psychiatric status of the three groups, and development was well balanced, corresponding to age. Protocols of the combined chemotherapy applied, and the incidence of anomalies, abnormalities, malignancies, and other diseases was recorded. Incidence was similar in all three groups. Incidence of congenital malformations was not increased in children conceived before and after therapy; however, a complex congenital abnormality, an atrial septal defect with horseshoe kidney, occurred in one young girl, conceived after the end of her father's treatment. The interval between conception and the end of therapy was established in the case of children conceived either during or after therapy. This was shorter in the case of healthy children; the number of healthy children conceived during cytostatic treatment was also remarkable. Further detailed analysis of data and individual evaluation of case reports are recommended. PMID- 8232079 TI - Wilms' tumor involving the inferior vena cava: preoperative evaluation and management. AB - Neoplastic invasion of the inferior vena cava due to renal tumors (especially Wilms' tumor) is uncommon in children. The tumor thrombus, according to the aggressiveness of the original neoplasm, can extend in diverse ways, obliterate the vascular lumen completely, and even reach the right atrium. The luminal thrombus might be accompanied by the involvement of the caval wall, which requires wide vascular resection. The purpose of this paper is to present our experience with 7 children, aged 18 months and 6 years, affected by caval invasion due to Wilms' tumor. Furthermore, the diagnostic techniques and the surgical treatment in simple caval thrombosis and in associated invasion of the caval wall are described. PMID- 8232080 TI - Malignant oligodendroglioma arising after radiation therapy for lymphoma. PMID- 8232081 TI - Right atrial thrombosis associated with central venous catheters in children with cancer. AB - Right atrial thrombosis (RAT) is infrequently diagnosed in children with cancer. Once RAT is documented, medical fibrinolysis or surgical thrombectomy is recommended. A RAT was documented in a child with lymphoma and was successfully lysed with recombinant tissue-type plasminogen activator. The case is presented and therapeutic options reviewed. PMID- 8232082 TI - Cervical epidural analgesia for a cancer child at home. AB - A cervical epidural catheter was placed in a 32-month-old girl suffering from painful bone metastases of a relapsing neuroblastoma in the left forearm. This therapeutic approach was chosen because oral morphine had failed to provide analgesia while producing intolerable side effects. The epidural injection of a mixture of morphine and bupivacaine provided complete analgesia with no side effects. This enabled the parents to take care of their child at home until the end, 35 days later. PMID- 8232083 TI - Total body hyperthermia in combination with etoposide and melphalan in a child with acute myelomonocytic leukemia. AB - In vitro and clinical studies have shown antineoplastic effects of hyperthermia alone and in combination with other treatment modalities. Synergistic cytotoxic effects of chemotherapy and hyperthermia have been demonstrated on leukemic cell clones in vitro. It seems that hyperthermia is effective in overcoming chemotherapy resistance. Several groups treated solid tumors by using total body hyperthermia (TBHT). However, only a few studies have been reported investigating the clinical effects of TBHT in myeloproliferative disorders. We report the case of a 7-year-old boy with myelomonocytic leukemia treated with TBHT (2 hours, 42 degrees C) combined with etoposide (600 mg/m2), melphalan (30 mg/m2) and hyperglycemia (200-300 mg/dl). Within 24 hours after TBHT, the leukemic cells decreased after TBHT from 53,000/microliters to zero. Skin leukemic infiltrates, resistant to conventional treatment, also responded well. Although our patient relapsed 34 days after TBHT, these results indicate that TBHT in combination with cytotoxic treatment may be a useful treatment modality in refractory leukemia. PMID- 8232084 TI - Gastric lymphoma in an 11-year-old boy: a case report. AB - A child with prolonged abdominal symptoms was shown to have a concomitant gastric non-Hodgkin lymphoma and gastritis caused by Helicobacter pylori infection. After a course of antibiotics and 6 months of chemotherapy, the patient is disease free. The relationship of the microbial infection and tumor development is discussed. The case supports a hypothesis that local H. pylori infection may be a predisposing factor for gastric lymphoma. PMID- 8232085 TI - High-dose methylprednisolone in resistant and relapsed children with acute lymphoblastic leukemia. PMID- 8232086 TI - Thymic cyst appearing after treatment of mediastinal non-Hodgkin lymphoma. AB - We report the first case of a thymic cyst appearing in the course of treatment for non-Hodgkin lymphoma of the anterior mediastinum. The patient was a 9-year old child in whom an abnormal contour of the left cardiac border persisted after chemotherapy, suggesting residual disease. The mass was found at thoracotomy to be a benign thymic cyst. The lesion was not present 2 years previously, and most likely represented cystic degeneration of the thymus, secondary to lymphomatous involvement. CT scan was not helpful in distinguishing the cystic lesion from residual lymphoma. PMID- 8232087 TI - [Guidelines for indications and implementation of intermittent artificial self ventilation. "Working Group for Home and Long-Term Ventilation"]. PMID- 8232088 TI - [How many patients with chronic viral hepatitis qualify for interferon therapy? Prospective analysis of university ambulatory care]. AB - How many patients with chronic viral hepatitis are suitable for interferon therapy? A prospective analysis of a university out-patient department. The present study prospectively analyzes 273 consecutive patients admitted to the outpatient-hepatology clinic for interferon therapy of chronic viral hepatitis from 1989 to 1991. 149/273 (54.6%) patients suffered from hepatitis-B viral infection (HBV). Of the 107 patients with NANB-hepatitis (39.2%) 103 patients finally proved to have hepatitis-C viral infection (HCV) (96.3%). In 17/273 patients (6.2%) the outside diagnosis of viral hepatitis was wrong. The percentage of men versus women was significantly greater in HBV (71.8% vs. 28.2%) than in HCV infection (52.4% vs. 47.5%) (p < 0.01; c2-test). The admittance of patients increased from 68 patients in 1989 to 88 patients in 1990 and 117 patients in 1991. The percentage of HCV-patients increased from 26.4% in 1989 to 38.6% in 1990 and 43.6% in 1991. The number of patients with HCV-infection admitted by non-university hospitals versus practitioners and internists was significantly greater in HCV-infection (48.5% vs. 51.5%) compared to HBV infection (hospitals: 36.9%; practitioners: 63.1%) (p < 0.05). Interferon therapy was initiated in 131/252 patients (52%) with HBV- or HCV-infection. HCV-patients were treated with interferon significantly more frequently (59.2%) compared with HBV-patients (46.9%) (p = 0.05). Since interferon therapy was initiated in only about 50% of HBV- and HCV-patients, the selection criteria for such treatment need to be publicized more effectively. The marked increase in patients with HCV infection suggests that recent improvements in serology of viral hepatitis are widely and appropriately used in clinical practice. PMID- 8232089 TI - [Data on the epidemiology of Crohn disease in the city of Cologne]. AB - In the period between 1.7.1985 and 30.6. 1986 283 patients with Crohn's disease were recorded prospectively in the city of Cologne, which has a total of 922,552 inhabitants. This corresponds to a prevalence of 30.67 per 100,000 inhabitants (women 35.97, men 24.90). During this one-year survey period, 47 new cases of the disease occurred, i.e. the incidence is 5.1 per 100,000 (women 5.8, men 4.3). The highest total incidence is in the age group of 20 to 24 years (17.3) and the highest total prevalence is in the group of 25 to 29 years of age (90.0). There is a substantial sex difference with a female/male ratio of 173:110. Only data of patients residing primarily in Cologne were evaluated. Figures are given on the diagnostic latency, the locations, operations, complications and misdiagnoses of Crohn's disease. PMID- 8232090 TI - [In vitro studies of the beat frequency of ciliary cell cultures after short-term exposure to SO2 and NO2]. AB - Mucociliary transport is an important defense mechanism of the respiratory tract. The aim of this study was to investigate the effect of SO2 and NO2 at different concentrations on ciliary beat frequency (ZSF). Single ciliated cells were obtained from 25 volunteers by nose brush. ZSF was quantified using video interference-microscopy. The cells were placed on a polycarbonate membrane, which was in contact with the surface of a reservoir filled with RPMI medium (bicarbonate buffered) or electrolyte solution (Ringer), allowing the cells to be supplied by capillarity. In an exposure chamber the cells were exposed for 30 to 120 min to SO2 2.5 to 15.0 ppm at 37 degrees C. SO2 induced a dose dependent decrease in ZSF of the cells, supported by Ringer solution. 2.5 ppm SO2 caused a 42.8%, 12.5 ppm a nearly 100% decrease (8.10 +/- 0.24 vs. 0.28 +/- 0.20 Hz). ZSF of cells cultured in RPMI medium was reduced moderately after 12.5 ppm SO2 exposure (7.90 +/- 0.26 vs. 6.66 +/- 0.31 Hz). In Ringer solution we observed a decrease of pH after 30 min SO2 exposure with 12.5 ppm to a minimum value of 3.6. In marked contrast, the pH of RPMI medium remained constant at 7.5 under identical conditions. After adding RPMI medium to Ringer solution, ZSF increased in parallel to the pH (5.0 ppm: 2.77 +/- 0.37 to 7.97 +/- 0.49 Hz). After an initial increase in ZSF, 120 min NO2 exposure to 15.0 ppm yielded a decrease in ZSF of 23.3% under conditions of constant pH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232091 TI - [Iodine deficiency in Germany]. AB - Urinary iodine excretion was determined in 2094 adults from 36 cities in the Federal Republic of Germany. The mean concentration was 94.1 +/- 93.1, median 66 micrograms iodine/g creatinine, or 6.2 +/- 6.0, median 4,4 iodine/dl urine, respectively. 194 (9.2%) subjects had an iodine excretion > 600 micrograms iodine/g creatinine or > 40 micrograms iodine/dl, which cannot be reached by normal nutrition. The median excretion found in West Germany (70.2 micrograms iodine/g creatinine and 5.0 iodine/dl urine, respectively) was higher than in East Germany (61.3 and 3.9, respectively), however the difference was not significant. There was no slope of urinary iodine excretion from North to South. The iodine creatinine index when compared with the iodine concentration alone did not improve the epidemiological accuracy (r = 0.75, p < 0.01). The iodine intake of the population is still below the minimum of the WHO recommendation. An efficient iodized salt prophylaxis would save approximately 1.3 billion DM annually and contribute to a genuine reduction in health-care costs. PMID- 8232092 TI - [Collagenous colitis]. PMID- 8232093 TI - [Cardiac diagnosis with conventional scintigraphy, positron emission tomography and magnetic resonance tomography]. PMID- 8232094 TI - [Aging and frailty--status of rights and legal responsibility]. PMID- 8232095 TI - [Functional obstruction of the upper airways as an expression of psychogenic vocal cord dysfunction. Fatal outcome of a functional disorder]. PMID- 8232096 TI - [Acute hepatorenal failure in hereditary fructose intolerance]. PMID- 8232097 TI - [Acute renal failure in HELLP syndrome]. PMID- 8232098 TI - Pathogenesis of lipid abnormalities in patients with nephrotic syndrome/proteinuria: clinical implications. AB - Hyperlipidaemia is an invariable complication of the nephrotic syndrome. The quantitative and qualitative changes in lipoproteins which occur may accelerate atherosclerosis. The pathogenetic mechanisms of the hyperlipidaemia are complex and poorly understood. Increases in lipoprotein production are compounded by reduced lipolysis of very low density lipoprotein and impaired catabolism of low density lipoprotein. Both proteinuria and hypoalbuminaemia have been implicated in the genesis of these abnormalities. The optimum treatment of the hyperlipidaemia has not been determined. 3-Hydroxy-3-methyl-glutaryl co-enzyme A reductase inhibitors appear to be the most effective lipid-lowering drugs, although their ability to reduce ischaemic events or prevent/delay renal failure remains to be proven. PMID- 8232099 TI - Proteinuria, hyperlipidemia, and the kidney. AB - Hyperlipidemia in the nephrotic syndrome is the result of abnormalities in both synthesis and catabolism of lipids and lipoproteins. The etiology of nephrotic hyperlipidemia has not been established, but both abnormal glomerular permeability to plasma proteins and reduced serum oncotic pressure may contribute. Although standard hypolipemic drugs are effective in nephrotic patients, therapies such as dietary protein restriction and angiotensin converting enzyme inhibitors which reduce proteinuria and increase serum oncotic pressure ameliorate hyperlipidemia as well. Hyperlipidemia may also induce proteinuric renal disease in normal animals and worsen renal injury in a variety of animal models of kidney disease. Conversely, treatment of hyperlipidemia prevents renal injury and lessens proteinuria. Potential mechanisms by which hyperlipidemia may cause renal injury include inflammatory and immunologically mediated injury and alteration of glomerular paracrine function. PMID- 8232100 TI - Relation of lipid abnormalities to progression of renal damage in essential hypertension, insulin-dependent and non insulin-dependent diabetes mellitus. AB - Lipid abnormalities are common in diabetes mellitus and in essential hypertension, but their contributions to diabetic and hypertensive nephropathy are unclear. Relevant data from several clinical trials are summarized here, as is a longitudinal study of diabetes mellitus showing that cholesterol correlates with both angiotensin II and blood pressure. The significant association of blood pressure and the renin-angiotensin system with declining renal function provides a mechanism whereby cholesterol may possibly impact adversely upon kidney function in diabetes. PMID- 8232101 TI - Intraglomerular lipid deposition in renal disease. AB - Lipid abnormalities are seen as important modulators of progressive renal disease. In most human glomerular diseases, localized accumulations of apolipoprotein (Apo) B- or Apo E-containing lipoproteins have been found in nonsclerotic glomeruli. Intraglomerular lipid deposition occurs in structurally damaged glomeruli, except for rare cases of dyslipoproteinemias. Studies of cases with lipid deposition in nonsclerotic glomeruli might provide important insights into the connection between lipids and the progression of original glomerular injury. PMID- 8232102 TI - Interactions between lipoproteins, glomerular cells and matrix. AB - Lipid deposition, mononuclear cell infiltration and accumulation of mesangial matrix components are recognized as early events in the development of glomerulosclerosis whilst correction of plasma lipid abnormalities slows the progression of renal disease in experimental models. In vitro studies have demonstrated that low density lipoprotein (LDL) is bound and internalized by mesangial cells, acts synergistically with growth factors to stimulate cellular proliferation and modifies secretion of chemotactic mediators and matrix components. LDL incubated with mesangial cells becomes oxidized and in this modified from inhibits cell proliferation and causes cytotoxic injury. Oxidation of LDL also modulates its effects on cell secretory function. Since proteoglycans secreted by mesangial cells bind LDL particles, excess matrix accumulation may exacerbate lipoprotein-mediated injury. These findings suggest that lipoproteins deposited and oxidized in the glomerulus may promote inflammation, cell injury and sclerosis. PMID- 8232103 TI - Renal hemodynamic consequences of hyperlipidemia. AB - Short-term dietary induced hypercholesterolemia in animals has been found to cause either increased vascular tone in various blood vessels, such as coronary arteries and renal vasculature, or impaired relaxation to endothelial-dependent vasodilators. In the kidney, renal blood flow, glomerular filtration rate and ultrafiltration coefficient are decreased whereas glomerular capillary pressure is elevated. These findings indicate pre- and post-glomerular constriction and mesangial cell contraction. Most evidence indicates that oxidized low density lipoprotein (ox-LDL) is responsible for initiating these functional abnormalities, and that it affects the endothelium rather than vascular smooth muscle cells. Nitric oxide (NO) and prostaglandin production are increased, probably due to the effects of ox-LDL on endothelial cell membranes. Vasoconstrictor prostaglandins and accelerated inactivation of NO by reactive oxygen species appear to play important roles in the impaired relaxation response. These endothelial abnormalities are postulated to contribute to progressive glomerulosclerosis via local hemodynamic changes and/or direct effects on mesangial cell proliferation and matrix synthesis. PMID- 8232104 TI - The mevalonate pathway: importance in mesangial cell biology and glomerular disease. AB - Products of intracellular mevalonate metabolism are critical for the growth and proliferation of eukaryotic cells. These products include cholesterol and several nonsterol isoprenoids. The isoprenoid farnesyl is a particularly important intermediate in the mevalonate pathway. Farnesyl can be used to synthesize cholesterol and can also bind covalently to several low molecular mass GTP binding proteins such as p21 ras. Farnesylated p21 ras may be critical for mitogenic signalling stimulated by growth factors such as platelet-derived growth factor. Inhibitors of the enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase, such as lovastatin and compactin, block the production of mevalonate and its metabolites. These agents have been shown to inhibit proliferation of many cell types. Recently we demonstrated that lovastatin inhibited proliferation of cultured glomerular mesangial cells. Lovastatin inhibition was overcome by the simultaneous addition of either mevalonate or farnesol, but not by exogenous low density lipoprotein cholesterol. These results suggested that farnesyl is critical for mesangial cell proliferation. In several experimental models of renal disease, chronic lovastatin administration reduced the extent of glomerular injury. The beneficial effects of lovastatin have been attributed to lowering of circulating lipid and lipoprotein levels. In view of recent data, however, it is possible that lovastatin may act to reduce glomerular injury, at least in part, through a direct action on mesangial cell proliferation. PMID- 8232105 TI - Lipid abnormalities and end-stage renal disease: implications for atherosclerotic cardiovascular disease? AB - Disorders of lipoprotein metabolism may have clinical implications as determinants of cardiovascular risk in patients with end-stage renal disease (ESRD). However, the role of hypertriglyceridemia or other parameters of lipoprotein metabolism as predictors of clinical measures of outcome during ESRD treatment is currently unknown. We determined the relationship between blood lipid, lipoprotein-cholesterol, apoprotein levels and the risk of cardiovascular death during 48 months of hemodialysis treatment in a prospective study of 129 patients with ESRD. Although serum triglyceride levels were increased in patients with preexisting cardiovascular disease, they were unrelated to clinical outcome during the period of follow-up. In contrast, lipoprotein(a) was an independent predictor of the risk of deaths attributable to cardiovascular disease during the prospective period of follow-up. Although these findings do not exclude the possibility that hypertriglyceridemic hemodialysis patients have atherogenic lipoproteins or associated metabolic conditions that predispose to cardiovascular death, our findings suggest that measurements of serum triglyceride concentration may not improve the cardiovascular risk assessment of hemodialysis patients. PMID- 8232106 TI - Risk factors for cardiovascular disease after renal transplantation. AB - The incidence of cardiovascular disease is high after renal transplantation. Risk factors include pretransplant cardiovascular disease, diabetes, male sex, serum cholesterol, hypertension, cigarette smoking, and allograft dysfunction. Prevention of cardiovascular disease can probably best be approached by modifying multiple risk factors. Thus, efforts should be made to identify and treat patients with severe coronary artery disease before transplantation. After transplantation, hypertension and hyperlipidemia should be treated, and efforts should be made to help patients abstain from smoking. Chronic immunosuppression protocols should attempt to balance the need to prevent allograft rejection with the known adverse effects of immunosuppressive agents on cardiovascular disease. Altogether, an aggressive, coordinated program to modify multiple risk factors is probably the best approach to the prevention of cardiovascular disease after renal transplantation. PMID- 8232107 TI - Treatment of hyperlipidemia in human renal disease. AB - Existing evidence suggests that dyslipidemia associated with long-lasting nephrotic syndrome and with chronic renal insufficiency may favor in the long run the occurrence of cardiovascular complications, and also aggravate glomerular damage with a pathological mechanism analogous to atherosclerosis. Correction of hypercholesterolemia and hypertriglyceridemia is therefore mandatory in both clinical conditions. This goal can be achieved with the combination of dietary intervention and the administration, even for long periods of time, of hypolipemic drugs (hydroxymethylglutaryl coenzyme A, HMGCoA, reductase inhibitors, to correct hypercholesterolemia in nephrotic syndrome, and fibric acids, to correct hypertriglyceridemia in uremic and dialyzed patients are the drugs of choice). In end-stage renal failure, the choice of the type of dialysis is also important. The value of extracorporeal LDL cholesterol removal is still to be proven. PMID- 8232108 TI - [Intestinal atresia: what is changed in the last 30 years]. AB - The changes occurred in the management of intestinal atresia in the last 30 years are presented. On the basis of a series of 55 neonates (29 M/26 F; 14 premature; mean body weight 2640 g, range 1340-4100 g; 23 duodenal, 30 small bowel, 2 colon) four aspects of this malformation are discussed; epidemiology, diagnostics, therapy and prognosis. Epidemiology. 40% incidence reduction is detected; unchanged the relative incidence of Down neonates with duodenal atresia. Diagnostics. Prenatal US scan useful in 4 of 8 cases only; emphasized the usefulness of plain X-ray abdomen. Perioperative treatment. Dramatic impact of the artificial nutrition on the management of short bowel syndrome, with 7 cases (residual small bowel from 75 to 9 cm, mean 40 cm) successfully treated. Surgical technique. Confirmed the validity of the one-stage termino-terminal anastomosis. Prognosis. Significant improvement in the last three decades respectively with 50%, 87% and 93% of long-term survivors. In the first decade 6 over 8 deaths were related to the atresia, while in the last 10 years the only death recorded was related to an associated severe cardiac malformation. PMID- 8232109 TI - [Epidemiologic and immunogenetic aspects of childhood diabetes mellitus]. PMID- 8232110 TI - [Neuropsychological development in children with focal brain injury]. AB - The study of children with focal brain injury has important implications from a clinical and theoretical perspective. Clinical data on children with congenital or early acquired lesions indicates that the cognitive sequelae are different from those resulting from similar damage sustained in adulthood. These differences depend in part on the differential effect that damage has on ongoing developmental process and in part on the different recovery capacity of the Central Nervous System of the child. From a theoretical perspective, focal lesion data is important for analyzing the issues of early neuropsychological functioning (especially in terms of early hemispheric specialization) and of plasticity and recovery of function of the CNS. This review analyzes the possible causes of this heterogeneity, that seems in part dependent on the interindividual variability of early neuropsychological organization and in part related to methodological factors such subject inclusion criteria and nature of neuropsychological measures. The review also discusses the role during development of the principal inter and intrahemispheric recovery mechanisms (special attention is given to intrahemispheric mechanisms which have been considered in the past as less determinant with respect to interhemispheric mechanisms). Furthermore, the role of lesion side as a prognostic parameter is discussed, specifically in terms of the evidence of a differential recovery capacity of left hemisphere with respect to the right. Various hypotheses have been put forward as possible interpretations of these data (maturational gradient, different neuropsychological organization of the two hemispheres), yet evidence is still controversial. If one considers the prognostic parameter--age of lesion onset--recent evidence does not confirm the hypothesis that the earlier the lesion, the greatest the recovery of function. Rather, it seems that relating age of lesion onset to other parameters, such as lesion side or size, has a greater and more reliable prognostic value. Another aim of the review is to analyze the effect that focal damage has on the dynamics of development. The fact that the effects of early damage might manifest themselves later in development, when the function in question is reaching a higher level of organization, underlines the importance of a longitudinal approach that assesses the developmental patterns of specific functions. Another issue discussed is the role of lesion side in determining specific cognitive impairment.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8232111 TI - [Social integration in adulthood in a group of subjects with Turner syndrome]. AB - The aim of the present study was to evaluate social integration in adulthood in a group of 48 subjects with Turner's syndrome. This was done by asking subjects to fill in a multi-choice questionnaire concerning their personal, social and working situation. The mean age (+/- SD) of the group was 24.8 +/- 1 and the reported stature was 142.2 +/- 1.4. The results obtained were compared with those formulated by ISTAT in 1990 for the entire Italian population. As far as regards education it was seen that 100% of subjects had completed primary school, 52% had attended secondary school, 29% had been to high school and 6% had attended university. It was therefore concluded that educational status, at least in this group, was higher than that of the Italian population in general. The cultural level of these subjects meant that most had found appropriate employment and only 6% were unemployed. 90% of the subjects were unmarried and only 5 were married (10%); the majority of those unmarried lived with their parents (83%). This underlines a prolonged dependence on the family nucleus and probable disorders regarding the subjects' own sexual identity and affective capacities. Among the parameters examined no substantial differences were found between subjects with a 45,X karyotype and those with chromosomic mosaicism. In the light of these findings it is apparent that efficacious medical and psychological strategies should be developed to enable a greater realization of interpersonal relations in the familial and social field. PMID- 8232112 TI - [Food intolerance and irritable bowel syndrome of childhood: clinical efficacy of oral sodium cromoglycate and elimination diet]. AB - Irritable bowel syndrome (IBS) is recognized to be a common cause of chronic diarrhea without failure to thrive in childhood. Several studies stressed the role of food intolerance as a major factor in the pathogenesis of IBS. The aim of this multicenter study was to investigate the offending role of food in IBS and to compare the therapeutic role of oral sodium cromoglycate versus elimination diet. 153 patients (mean age 4 years) with diarrhea (> 3 stools per day for four days in a week) and abdominal pain for about 10 months were enrolled in this trial. About half of the patients had a family history positive for atopy and 70% of the cases complained of intestinal symptoms after food ingestion. In 17% of the patients Skin Prick test (SPT) resulted positive to at least one food allergen and 87% of positive reactions to SPT was provoked by common foodstuffs. 87% of patients treated with elimination diet (rice, lamb, turkey, lettuce, carrots, sweet potatoes, pears, oil, tea, salt, mineral water, brown sugar) and 97% of patients treated with SCG (mean 63 mg/kg/day) for one month showed a significant improvement of intestinal symptoms. An elimination diet for several weeks can produce, beside a bad compliance (23% of patients admitted to our study didn't strictly follow diet regimen) also a nutritional deprivation. The results of this trial suggest that it's correct to investigate the role of food in children with diarrhea not due to organic diseases and diagnosed such as IBS and to use oral SCG to obtain the improvement of these symptoms. PMID- 8232113 TI - [Discitis in children. Report of two cases]. AB - Two cases of intervertebral disk infection in 2 3-year-old boys are presented. The study reported on the anamnesis, clinical, laboratory and instrumental findings as well as therapy in both cases. Diagnosis was suspected on the basis of pain, refusal to assume any position that flexes the spine and irritability. Low-grade fever, the moderate increment of inflammatory indexes and the characteristic changes seen in 1 case on Magnetic Resonance Imaging (MRI) only and in the other one on both Technetium 99m polyphosphate bone-scanning and MRI verified the clinical diagnosis. Furthermore the authors briefs discuss the differential diagnosis of disc-space inflammation and several acute and subacute diseases of infancy and childhood some of them not localized in the spine (myopathy, appendicitis, peritonitis, urinary infections) for which discitis may be often mistaken. PMID- 8232114 TI - [Sudden infant death syndrome. A case with accessory atrio-ventricular pathways and fetal ring tissue remnants]. AB - A 55 day-old male infant dying suddenly is diagnosed as sudden infant death syndrome (SIDS). Important modifications of the cardiac conduction system were found; such as: splitting-His bundle dispersion, accessory atrioventricular pathways of Mahaim type, and remnants of fetal "ring tissue" anastomosing with ordinary myocardium. These changes can be considered as arrhythmogenic in nature. PMID- 8232115 TI - [On Apert syndrome. A case report]. AB - The authors, after a description of Apert syndrome, have reported a case of an observed newborn. In this case the paternal age was not determinant and the mother was young and in good health. No chromosomal anomalies were found as to the baby. They have reported the baby's malformative condition has been reported in order to give a contribution that may increase the cases observed worldwide. PMID- 8232116 TI - [Virilization in women. Clinical and therapeutic aspects]. AB - Androgenization in women can be divided, from a clinical standpoint, in two groups: a major form (with hirsutism, seborrhea, acne, hair loss, menstrual irregularities, masculinization of muscles and voice, mammary atrophy) and a minor one, with skin changes only (in particular hirsutism) with or without menstrual problems. The different clinical presentations are reviewed here: virilizing tumours of adrenal glands and ovaries, adrenogenital congenital syndromes, Cushing's syndrome and disease, iatrogenic forms, simple or idiopathic hirsutism, late onset enzymatic defects of adrenal steroidogenesis, polycystic ovary syndrome). The relevant therapeutic options are discussed. Special attention is devoted to the treatment of simple cutaneous androgenization, a problem affecting about 10% of women, by antiandrogenic drugs, mostly cyproterone acetate and spironolactone. These compounds compete with dehydrotestosterone for androgen cutaneous receptors, and have obtained good results, although not permanent. The indications, use and side-effects are also discussed. PMID- 8232117 TI - [Probable role of obesity on the adrenal response to acute stimulation with adrenocorticotrophic hormone in eumenorrheic and hirsute, non-eumenorrheic women]. AB - Obesity, the most frequent nutritional problem throughout the rich nations, can have a vast and significant influence on different aspects of endocrinology, in particular on ovulation disfunction, on hyperandrogenemia, on hormone-sensitive carcinomas. Our study proposes to value the response to adrenal cortex to stimulation with adrenocorticotropin (ACTH) hormone in obese patients, with particular attention to the behavior of adrenocortical androgens and their precursor. We recruited 30 female patients so divided: 12 obese, nonhirsute, eumenorrheic patients (group A); 10 normal weight, hirsute patients in situation of secondary amenorrhea (group B); 8 normal weight, nonhirsute, eumenorrheic patients (group C). Cortisol, progesterone, 17 OH progesterone, dehydroepiandrosterone sulfate, androstenedione, testosterone were measured at 60, 120, 180, 240, 300 min during continual infusion i.v., for 5 h, of ACTH 1-17 at 100 mcg dose, in physiological sodium chloride solution. All the women with monthly menstruation were studied between the IV and VIII day of their cycle. In the patients with secondary amenorrhea the value of basic progesterone was used to completely exclude an eventual luteal phase and the relationship LH/FSH was so as to logically exclude a diagnosis of polycystic ovary. This exclusion was also confirmed from the report of the ultrasonography. The basic concentration of hormone dosage is not significantly different between the patients of the three groups, except for T. This hormone is different because it is found to be significantly (p < 0.01) increase in the hirsute patients, in respect of the patients in group A and group C. Also P and 17OHP have been found to be higher, if only in insignificant measure, in hirsute patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232118 TI - [Transverse study of obesity: distribution of adipose tissue and correlated pathology]. AB - The relationship between obesity and prevalence of dyslipidemia is well known. Recent studies affirm that differences in fat distribution can be predictive for differences in the prevalence of metabolic disturbances and cardiovascular disease independently of the BMI, presently the most common index of obesity. In order to verify whether body fat distribution can be associated with a higher risk of atherosclerosis, we have evaluated in a group of obese women the eventual presence of endocrine and metabolic diseases. Assessing regional fat distribution, the waist/hip ratio has been shown to be more closely correlated with these diseases than BMI. We have studied two groups of 10 women, comparable for age and BMI: group A aged 45.8 +/- 6.9 years with a BMI of 35.6 +/- 2.8 kg/m2; group B aged 48.3 +/- 3.6 years with a BMI of 38.5 +/- 2.8 kg/m2. The women were divided according to the waist-hip ratio, which was calculated by measuring the circumference of the waist, namely the smallest circumference between the xiphoid and the umbilicus, and the circumference of the hips at the point of the maximum protuberance of the buttocks. The cut-off value for the waist/hip ratio was considered as 0.80 for the reason that this variable is the most accurate cut-off value for abdominal obesity: for group A 0.76 +/- 0.02; for group B 0.89 +/- 0.02 (p < 0.01). All the women were healthy. None of them was in therapy with any kind of drugs, nor was there any restriction to diet. Nobody was a smoker, neither did anyone drink alcoholic beverages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232119 TI - [Hashimoto's thyroiditis presenting as a "hot nodule"]. AB - Hashimoto thyroiditis is clinically the most variable thyroid disease. We describe a case presenting as "hot nodule". This unusual scintigraphic appearance and its possible explanation are discussed. PMID- 8232120 TI - [Fluoxetine and obesity]. PMID- 8232121 TI - [Measurement of the association between hyperinsulinism and arterial hypertension in obese subjects]. PMID- 8232122 TI - When luck runs out. Compulsive gambling: bad habit or serious illness? PMID- 8232123 TI - Improving communication between physicians and Medicare carriers. PMID- 8232124 TI - Acute pyogenic spondylodiscitis with epidural phlegmon. Diagnosis and management by MRI and multidisciplinary approach. PMID- 8232125 TI - Regulating physician conflicts of interest. Tighter restrictions on referral relationships among providers. PMID- 8232126 TI - Confidentiality of medical records under Minnesota law. PMID- 8232127 TI - Long-term care insurance. Putting a premium on peace of mind in retirement. PMID- 8232128 TI - Maintaining the public trust. Physicians must put patients' interests first. PMID- 8232129 TI - Compulsive gambling. When the stakes are too high. PMID- 8232130 TI - Business prescription. Can Winston Wallin cure U' health sciences' ills? PMID- 8232131 TI - When one of 'us' has cancer. PMID- 8232132 TI - [The quantitative assessment of DNA in potentially cancerous cases of oral lichen]. AB - The increased prevalence of all variants of oral lichen, both as a result of increasing frequency and improved knowledge of the pathologist, coupled with its greater trend to malignant transformation has focused the attention of researchers on the development of new technology that could help in the early detection of the precancerous lesion. DNA cytometric detection can be useful to detect the precancerous lesion when clinical and histological findings of the transformation are still absent. The early diagnosis of such lesions entitles to use a more aggressive treatment, both medical and surgical. PMID- 8232133 TI - [The psychopathological aspects of oral lichen planus (OLP)]. AB - The etiopathology of oral lichen planus (OLP) is still uncertain. It is probably caused by a T lymphocytes cell-mediated autoimmune reaction. This reaction, caused by different pathogenic noxae, develops in subjects with a specific genetic pattern (HLA-DR on the keratinocytes) and some psychic attitude (higher Hamilton anxiety scale (HAS) and Hamilton depression scale (HDS) scores. In literature a psychic component has been often mentioned in OLP etiopathology. This work is a case-control study of replication: it aims to evaluate the importance of psychopathologic trait in patients affected by OLP. The study has been carried out on 16 patients (7 male and 9 female--average age 56) with OLP histologically confirmed. Before biopsy, they had been tested with: General health questionnaire (GHQ). Hamilton anxiety scale (HAS). Hamilton depression scale (HDS) with melancholia scale (MES). We have also studied a survey group of 14 patients affected by leukoplakia and neoplasms of the oral cavity (12 males and 2 female--average age 51). Before biopsy the survey group has been tested with the same questionnaire faced by the studied group. We chose the survey group as follows. Patients with leukoplakia show such similar lesions to OLP that the diagnosis is often histological; as far as patients with oral cavity tumours are concerned we hypothesized a more than usual anxious and depressed behaviour. We compared the score of the OLP patients to the one of the survey group patients using the "One-way Analysis of Variance" (ANOVA) method. The results confirmed that the patients with OLP showed very high scores in HAS and HDS, while the survey group scores were significantly lower.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232134 TI - [Pharmacological support during orthodontic therapy with a topical anti inflammatory]. AB - On the occasion of many orthodontic procedures is often necessary to use NSAIDs in order to provide relief from the inflammatory and painful symptoms originating by removable and permanent devices. In this single blind controlled study, efficacy and tolerability of a topical antiinflammatory drug, formulated as mouth wash containing flurbiprofen 0.25%, have been evaluated in 60 patients (range 12 16 years) suffering from tissular injuries caused by permanent devices. Following a randomisation list, the patients were allocated to 3 different balanced groups, the first received flurbiprofen mouth wash 10 ml, 3 times daily for 7 days, the second received placebo mouth wash in the same way and the third group was a control one. The aim of the trial was to evaluate the pain symptom related to the injuries. The result of the study shows statistical improvement in favour of flurbiprofen in comparison to the placebo as well as to the control group. No local or systemic ADRs were reported. PMID- 8232135 TI - [The diagnosis and surgical therapy of maxillofacial malformations. The statistical considerations]. AB - In this report the authors develop a thorough statistic analysis upon the past 12 year activity (1980-1991) of the University of Rome, Department of Maxillo-Facial Surgery, in the field of the diagnosis and treatment of congenital or acquired maxillofacial malformations. They point out, in particular, the high frequency rate of bimaxillary osteotomy use in the surgical treatment of dentoskeletal malformations, that are the most frequent malformations in maxillofacial clinical practice. They also point out how wide the use of the rigid internal fixation technique in this surgical field has become. PMID- 8232136 TI - [An analysis of the prevalence of isolated dental anomalies and of those associated with hereditary syndromes: a model for evaluating the genetic control of the dentition characteristics]. AB - Tooth ontogenesis, morphogenesis, and eruption are under a multifactorial (genetic, epigenetic, environmental) control. The aim of this study has been a genetic appraisal of tooth developmental characteristics by means of the prevalence and patterns of association of tooth anomalies in a large sample of hereditary syndromes. MATERIALS AND METHODS. A sample of 169 hereditary syndromes exhibiting tooth abnormalities was collected from the literature: original articles were controlled in order to achieve uniformity of judgement about the definition of the dental defects. Tooth anomalies were classified according to anatomoclinical criteria: number (hypodontia/hyperdontia), shape, position, structure (enamel/dentin/cementum), size (micro-/macrodontism), eruption (precocious/delayed including unerupted teeth). Prevalence data were calculated for: 1) modes of syndrome inheritance; 2) syndromes exhibiting a single dental anomaly vs. syndromes exhibiting associated tooth abnormalities; 3) different types of tooth abnormalities; 4) isolated and associated tooth abnormalities. PMID- 8232137 TI - [The therapy of anterior disk dislocation in craniomandibular disorders. A clinical case report]. AB - The authors discuss the outcomes of therapy for CMDs and present a clinical case reporting an alternative method for the treatment of disc displacement. The orthopaedic therapy for CMDs is based on the use of interocclusal splints. Two kind of splints are prescribed to patients with CMDs: stabilization or repositioning splint. The former is used for a conservative treatment, the latter for irreversible treatment. In view of the poor long-term success of repositioning therapy, the cost of subsequent dental treatment, and the great potential for iatrogenic occlusal and muscular problems, the authors believe that the initial treatment approaches should be, in any case, conservative, reversible, and non invasive. The authors prescribe to their patients the repositioning splint for use either part-time or full time use. Part-time use is effective for preventing disc displacement without reduction. If used full time, once joint noises, pain, and displacement are decreased, the appliance should be adjusted to/or replaced with a stabilization appliance to allow posterior positioning of the mandible into a more physiologically stable position. The authors present a clinical case, a 27 yo female with disc displacement with reduction was treated with stabilization splint. After 6 weeks of therapy she returned reporting a temporary closed lock and sharp pain in the left TMJ. An anterior repositioning splint was fabricated in order to position the mandible forward. After 10 weeks of this treatment the patient reported absence of joint/muscle symptoms and of joint noise.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232138 TI - Clinical aspects and microbiology of HIV-associated periodontal lesions. AB - This study examines the clinical aspects of HIV-associated periodontal lesions and the prevalence of periodontal pathogens. Subgingival plaque samples were taken from 55 subjects in six study groups: 1) HIV-seropositive patients with gingivitis, 2) necrotizing gingivitis, 3) periodontitis or 4) with health periodontium, 5) patients with rapidly progressive periodontitis or 6) periodontally healthy in whom there was no evidence of HIV infection. Among HIV positive patients there was a majority (66%) of intravenous drug users. We detected more Bacteroides intermedius, B. buccae and B. oralis in HIV-infected patients with periodontal lesions and in HIV-negative subjects with rapidly progressive periodontitis than in the other groups. High levels of Spirochetes were recovered in both HIV associated necrotizing gingivitis and periodontitis. The results indicate that there is a similarity in the microbiological profile of HIV-associated gingivitis, necrotizing gingivitis, periodontitis and rapidly progressive periodontitis of HIV-negative subjects although significant differences in the clinical aspects of the lesions and in the immune status of the host have been observed. PMID- 8232139 TI - Theory and practice: beyond the dichotomy. AB - Equipping students with the means to bring theoretical understanding to bear on the implementation of practical nursing skills has been an enduring problem for nurse educators. The problem is partly born out of a clinical culture in which the 'merely theoretical' tends to be dismissed. The aim of this paper is to show ways of thinking about theory and practice which actually avoid fruitless dichotomy. Theoretical reflection and practical action are in reality richly interconnected (Heidegger 1962). In the case of the expert practitioner (Benner 1984) theory and practice may be impossible to distinguish. New thinking on this problem is particularly urgent in the light of the Project 2000 initiative. It is hard to see how this new approach to nurse education can be entirely successful if students are allowed to get into the way of contrasting the reality of practice with the 'merely theoretical'. PMID- 8232140 TI - In defence of pedagogy: a critique of the notion of andragogy. AB - Malcolm Knowles' theory of andragogy has gained increasing acceptance among nurse educators. Andragogy is espoused as a progressive educational theory, adopted as a theoretical underpinning for curricula and is even considered to be synonymous with a variety of teaching techniques and strategies such as 'problem-based' and 'self-directed' learning. This paper offers a critique of the notion of andragogy which maintains that the distinction created between andragogy and pedagogy is spurious and based upon assumptions which are untenable. It is argued that andragogy has been uncritically accepted within nursing education in much the same way that the nursing process and models of nursing were in their day. Finally, it is claimed that true pedagogy has far more radical, powerful and transformative possibilities for nursing education. PMID- 8232141 TI - Stressors to student nurses beginning tertiary education: an Australian study. AB - Stress placed upon student nurses entering tertiary education during a period of rapid change in the Australian system was examined. Overall findings were consistent with research elsewhere, for example, regarding students' experience of anxiety created in particular through assessment requirements. Older students experienced greater anxiety than others in relation to the science components of the curriculum; other groups were also found to experience particular problems. Changes in course design and appropriate support strategies which reduce stress for beginning students are likely to contribute to an improvement in student performance and a decrease in student attrition, and are deemed to be essential if nursing is truly to be viewed as a caring profession. PMID- 8232142 TI - Supervision as a focal point in nurse education: an exploratory study of its effects on the needs of student nurses. AB - This study reports the results of a 2 year follow up of the effects of experience of supervision on the personal needs of nursing students. Supervision was given during this period on a continual basis as a focal point of nursing education. Students were encouraged to relate critical incidents as starting points for the supervisory process. Students rated the quality of supervision in terms of its perceived effect on their sense of professional identity and on their self confidence with respect to clinical work. A Swedish version of a standard personality questionnaire, the Edwards Personal Preference Schedule (EPPS), was given at the initiation of supervision and repeated 2 years later on its completion. The results of 48 students are presented. Two hypotheses relating to the global effects of supervision were not confirmed but the hypothesis concerning the effects of supervision to depend on its perceived quality was confirmed. As measured, the effectiveness of supervision was seen in statistically significant correlations between ratings of supervisory quality and changes reflecting an increased need for leadership and a diminished need for subordinance. PMID- 8232143 TI - Exploring the process of learning in human ecology. AB - The process of learning phenomenon has been and is still widely discussed in nursing education especially in relation to adult learning. In this paper, the author reflects upon and analyses his own specific learning activity in the human ecology perspective to construct a conceptual framework of how learning takes place. He begins by briefly defining human ecology within which this learning process takes place and explores in general terms the existing definitions of 'learning' before arriving at his working definition. He then relates, and explores his learning experience as a basis for analysing how learning actually takes place and uses the identified learning characteristics within this mode of learning to support its relevance and implications for nurse education. PMID- 8232144 TI - From common foundation programme to branch: recognising a status transition. AB - The transition from the Common Foundation Programme to the Branch programmes in the Diploma in Nursing Studies course (Project 2000) is described as a major status passage. While there are many positive aspects to this transition, particular areas of concern are highlighted in the hope that these will be addressed. PMID- 8232145 TI - The content of AIDS counselling workshops for nursing students. AB - The need to train nurses to care for people with acquired immune deficiency syndrome (AIDS) is an important one. The need, also, to help them to develop counselling skills in this field is also vital. This paper reports the responses of 21 workers in the AIDS field (including nurse educators and teachers) to the question: 'If you were setting up an AIDS counselling course for nurses, what sorts of elements would you have in it?' The responses were content analysed and grouped into 8 categories. These were: information about AIDS; psychology and sociology; attitudinal issues; sexuality; personal issues; counselling; visits from other people and the history of AIDS. The issues within these 8 categories of content for an AIDS counselling skills course for nurses are discussed. PMID- 8232146 TI - A career in nursing for the physically disabled individual? AB - This paper highlights the small percentage of nurses who are disabled, and seeks to give some explanations for this phenomenon by reference to theories of stigma and labelling. Having acknowledged that one of the main barriers to disabled people entering nursing may be an attitudinal one on the part of health care staff, some recommendations are made in relation to nursing curricula, which may facilitate the recruitment and education of people with physical disabilities. PMID- 8232147 TI - Dealing with child abuse: development and evaluation of a short workshop for nurses. AB - A 1-day workshop was evaluated in terms of its effectiveness in enhancing nurse (N = 70) understanding of child abuse. Measures of their knowledge and attitudes concerning abuse were administered before and after the workshop. The results indicated that the workshop led to a significant improvement in these two areas, in particular, a sub-sample (N = 25) who received an additional teaching aid (a programmed learning script) did better than their colleagues. Finally, a teaching evaluation form indicated a high degree of consumer satisfaction. It is concluded that the brief workshop was successful in enhancing knowledge and attitudes, and that it illustrates the kind of 'professional approach' recently advocated in the literature. PMID- 8232148 TI - Violence in America: an issue of public health. PMID- 8232149 TI - Let's not have a medical Chernobyl. PMID- 8232150 TI - Dermatology quiz #7. Pyogenic granuloma. PMID- 8232151 TI - Handling patient complaints. PMID- 8232152 TI - Computerized medical records. Dream or reality? AB - For over 25 years many clinicians and others working in health-related fields have envisioned the day when physicians will use a computer on their desk or at the bedside to enter and retrieve the medical records and other information needed to care for their patients. Pioneers in medical informatics have been working during this time to develop the concepts and systems to make this possible, but so far, with only a few exceptions, this has been still mostly a dream. However, we may now be on the threshold of the time when these systems will be implemented on a large scale and become an integral part of the practice of clinical medicine, making the vision no longer a dream but a reality. PMID- 8232153 TI - Consumption of fruits and vegetables in Missouri. AB - High consumption of fruits and vegetables has been linked with a reduced risk of several important chronic diseases. The authors utilized telephone survey techniques to assess the level of fruit and vegetable consumption among adult Missouri residents. Only 28% of respondents reported consumption of five servings of fruits and vegetables per day. Older females had the highest rate of consuming five servings per day, while young males had the lowest rate. These findings, in conjunction with national data, show the need for increasing emphasis on the "5 A Day" campaign. PMID- 8232154 TI - Biliary cystadenoma: a case report and review of the literature. AB - Biliary cystadenoma is a rare benign neoplasm usually found in the parenchyma of the right lobe of the liver. The lesions are usually septated and represent less than 5 percent of non-parasitic cysts of biliary origin. Malignant degeneration to biliary cystadenocarcinoma is well documented, and recurrence is the rule following incomplete excision. PMID- 8232155 TI - Cholelithiasis: seven critical questions. AB - The management of cholelithiasis is usually straightforward, but there are a few special situations and questions which arise repeatedly. This paper deals with management of patients with asymptomatic stones, diabetic patients with stones, the relationships between stones and cancer, and patients without stones but who have typical pain. It also considers certain surgical topics of interest to primary care specialists, including that of safety of laparoscopic surgery, pain after surgery, and the role of incidental cholecystectomy. PMID- 8232156 TI - The flood of '93. PMID- 8232157 TI - The name game: getting into those "impossible to get into" colleges. PMID- 8232158 TI - Data is driving health care decisions ... be wise--know your data. PMID- 8232159 TI - The use of rectal temperature to monitor heat stroke. PMID- 8232160 TI - Variations in three cardiovascular disease risk factors in Missouri, 1986-1992. AB - In 1991, cardiovascular disease claimed the lives of over 22,000 Missourians. Recent studies show that much of this mortality can be eliminated through risk factor reduction and early detection. This article examines the regional and demographic differences in three major cardiovascular disease risk factors and outlines some steps that physicians and other health care providers can take to reduce their prevalence. PMID- 8232161 TI - Cutaneous mycosis: sporotrichosis. PMID- 8232162 TI - Computer-assisted instruction. PMID- 8232163 TI - Let us now revisit prepaid group practice. AB - American medicine will soon enter health care reform. The Clinton Administration's Task Force on Health Care Reform is likely to introduce change that relies heavily on managed care, a field now dominated by the health insurance industry. Physicians can manage health care themselves by forming Prepaid Group Practices (PGPs). PGPs combine health insurance functions with physician managed health care. They can negotiate directly with health care purchasers, thereby abrogating the commercial health insurance industry. PMID- 8232164 TI - Making sense of coagulation tests, a practical approach. AB - A large variety of "clotting studies" are available to the clinician in evaluating the hemostatic capability of his patient. The authors have experience with these tests which are available in varying sophistication at hospitals ranging from 25-600 beds. In this article, the biochemistry and physiology of coagulation testing are purposely avoided. A common sense approach to evaluating the patient's "chance of bleeding or clotting" is presented. PMID- 8232165 TI - An adult case of acute biphenotypic leukemia with characteristic mixed morphology. AB - A case of acute biphenotypic leukemia with mixed blast morphology and combined myeloid and T-lymphoid features is reported. The leukemic cells consisted of small lymphoid hand-mirror blasts and large blasts with cytoplasmic granules and rare Auer rods. The cells expressed myeloid and immature T-lymphoid features by cytochemistry and immunophenotyping, however T cell receptor genes were in germline configuration. Cases of biphenotypic leukemia with similar morphological and immunophenotypic findings have been described previously in children. This case represents a morphologically and phenotypically distinct subtype of acute biphenotypic leukemia. PMID- 8232166 TI - The changing image of the American physician. PMID- 8232167 TI - Physician and other health-care professional counseling of smokers to quit- United States, 1991. AB - Physicians and other health-care professionals play a lead role in the prevention of tobacco smoking in the United States. In particular, health-care professionals can assist patients to stop smoking by counseling them about quitting. To monitor progress toward the national health objectives for the year 2000 on tobacco use, data from CDC's 1991 National Health Interview Survey-Health Promotion and Disease Prevention (NHIS-HPDP) supplement were used to estimate the prevalence of outpatient physician and other health-care professional counseling of smokers to quit. This report summarizes the results of that survey. PMID- 8232168 TI - Mortality trends for selected smoking-related cancers and breast cancer--United States, 1950-1990. AB - During 1990, nearly 419,000 deaths (approximately 20% of all deaths) in the United States were attributed to smoking, including more than 150,000 deaths from neoplasms. Cigarette smoking remains the single most preventable cause of premature death in the United States. Based on current and past smoking patterns, the public health burden of smoking-related cancers is expected to continue during the next several decades. The death rate for smoking-related cancers varies by race; race reflects differing distributions of several risk factors for smoking-related cancers (e.g., high-risk behaviors) and is useful for identifying groups at greatest risk for smoking-related cancers. This report describes mortality trends for cancers (i.e., lung, oral cavity and pharynx, esophagus, and larynx) that are at least 70% attributable to smoking and other tobacco use (2) by race and sex. In addition, because lung cancer recently surpassed breast cancer as the leading cause of cancer deaths among women, death rates for lung cancer are compared with those for breast cancer. PMID- 8232169 TI - Update: mortality attributable to HIV infection among persons aged 25-44 years- United States, 1991 and 1992. AB - During the 1980s, human immunodeficiency virus (HIV) infection emerged as a leading cause of death in the United States (1). In 1992, HIV infection became the number one cause of death among men aged 25-44 years. This report updates national trends in deaths caused by HIV infection for 1991 and 1992. PMID- 8232170 TI - Assessment of street outreach for HIV prevention--selected sites, 1991-1993. AB - Street outreach programs for human immunodeficiency virus (HIV) prevention are designed to deliver HIV prevention messages, materials, and referral services to high-risk persons outside of traditional health-care and drug-treatment clinics. The Acquired Immunodeficiency Syndrome (AIDS) Evaluation of Street Outreach Projects (AESOP) is an eight-site study designed by CDC in collaboration with researchers in each of the sites to better understand client characteristics, service delivery, and the impact of street outreach programs on the risk behaviors of high-risk populations. The populations studied are injecting-drug users (IDUs) in five of the eight sites and youth in high-risk situations (YHRS) (i.e., youths aged 12-23 years who are homeless or runaway or who support themselves through the "street economy" of drugs, prostitution, pan-handling, and crime) in three sites. This report summarizes information collected during the first 2 years of the project. PMID- 8232171 TI - Distribution of STD clinic patients along a stages-of-behavioral-change continuum -selected sites, 1993. AB - Human immunodeficiency virus (HIV) counseling and testing are important components of the public health effort to contain the acquired immunodeficiency syndrome (AIDS) epidemic. Project RESPECT is a multicenter randomized trial being conducted by CDC at five sexually transmitted disease (STD) clinics to evaluate the efficacy of HIV counseling interventions in preventing HIV and other STDs. Because consistent and proper use of latex condoms is one method to substantially reduce the risk for transmitting HIV and other STDs (1), the HIV prevention interventions being evaluated focus primarily on increasing consistent condom use with primary and other sex partners during vaginal and anal sex. Before initiating the randomized trial, pilot studies were conducted to better characterize the population and to develop an operationally feasible enhanced intervention. This report describes the results of one pilot study. PMID- 8232172 TI - Severe acute respiratory illness linked to use of shoe sprays--Colorado, November 1993. AB - On November 3, 1993, the Colorado Department of Health (CDH) was notified of acute respiratory illness in a Colorado woman following use of an aerosolized leather-shoe conditioner. Active surveillance by CDH identified two additional cases. This report summarizes the case investigations. PMID- 8232173 TI - Dental health of school children--Oregon, 1991-92. AB - Dental caries remains among the most prevalent diseases of both children and adults. To establish a baseline for monitoring oral disease trends in Oregon, the State Health Division, Oregon Department of Human Resources; Oregon Health Sciences University; and Multnomah County Health Department collaborated in a statewide assessment or oral health needs. Phase 1 (1991-92) evaluated Head Start and elementary school children. Phase 2 (1993) is assessing the oral health of adults. This report presents the results of Phase 1. PMID- 8232174 TI - Mortality patterns--United States, 1991. AB - In 1991, 2,169,518 deaths were registered in the United States--21,055 more than in 1990 and the most ever recorded. Despite this increase, the overall age adjusted death rate was 513.7 per 100,000 population, the lowest ever recorded. Provisional data for 1992 indicate that the death rate continued to decline through 1992. As in previous years, nearly two thirds of deaths in 1991 were caused by the first three leading causes of death (i.e., heart disease, cancer, and stroke). This report summarizes mortality data for 1991 and compares patterns with 1990. PMID- 8232175 TI - Reduction in alcohol-related traffic fatalities--United States, 1990-1992. AB - Alcohol-related traffic crashes are a leading cause of unintentional injury deaths and a substantial contributor to health-care costs in the United States. Approximately 40% of persons will be involved in an alcohol-related crash during their lifetime. In 1992, alcohol was involved in an estimated 17,700 traffic fatalities and 355,000 traffic injuries. In 1990, alcohol-related crashes cost $46.1 billion, including $5.1 billion in medical expenses. This report summarizes data regarding alcohol-related traffic fatalities (ARTFs) from the National Highway Traffic Safety Administration's (NHTSA) Fatal Accident Reporting System (FARS) during 1982-1992. PMID- 8232176 TI - Update: influenza activity--United States and Europe, 1993-94 season. AB - In collaboration with the World Health Organization (WHO) international collaborating laboratories and with state and local health departments in the United States, CDC conducts surveillance to monitor influenza activity and to detect antigenic changes in the circulating strains of influenza viruses. Laboratory surveillance indicates the predominance of influenza type A so far this season. This report summarizes influenza activity in the United States and Europe from mid-September through mid-November 1993. PMID- 8232177 TI - Hypothermia-related deaths--Cook County, Illinois, November 1992-March 1993. AB - Hypothermia results from the inability of the body to maintain a sufficiently high body temperature and is defined clinically as the lowering of core body temperature to < or = 95 F (< or = 35 C). Environmental hypothermia results from a combination of heat loss by convection (degree of wind exposure), conduction, and radiation to the surrounding air. Although hypothermia-related deaths are preventable, during 1979-1990, 9362 deaths in the United States were attributed to environmental hypothermia or excessive cold (International Classification of Diseases, Ninth Revision [ICD-9], codes E901.0, E901.8, and E901.9; excludes manmade cold [E901.1]). From November 1992 through March 1993, 22 hypothermia related deaths were identified by the Cook County (Chicago), Illinois (1990 population: 5,105,067), medical examiner. This report summarizes information on those deaths and describes specific findings in four of the deaths. PMID- 8232178 TI - Jin bu huan toxicity in adults--Los Angeles, 1993. AB - Jin Bu Huan (JBH) is a traditional Chinese herbal product used as a sedative and analgesic. During 1993, public health and health-care providers in Colorado reported three children with unintentional overdoses of JBH that caused central nervous system and respiratory depression with rapid onset of life-threatening bradycardia. Subsequently, the first cases of acute hepatitis attributed to use of JBH were diagnosed in three women in Los Angeles during July and August 1993. Patients 1 and 2 were referred to a Los Angeles hepatology clinic by their physicians; patient 3 was identified by patient 2. All three patients had purchased JBH at the same health-food store. This report summarizes the investigation of these cases. PMID- 8232179 TI - Surveillance for waterborne disease outbreaks--United States, 1991-1992. AB - PROBLEM/CONDITION: Since 1971, CDC and the U.S. Environmental Protection Agency have maintained a collaborative surveillance program for collection and periodic reporting of data on the occurrence and causes of waterborne disease outbreaks. REPORTING PERIOD COVERED: January 1991 through December 1992. DESCRIPTION OF SYSTEM: The surveillance system includes data about outbreaks associated with water intended for drinking and also about those associated with recreational water. State and local public health departments are the agencies with primary responsibility for the detection and investigation of outbreaks. State and territorial health departments report these outbreaks to CDC on a standard form. RESULTS: For the 2-year period 1991-1992, 17 states and territories reported 34 outbreaks associated with water intended for drinking. The outbreaks caused an estimated 17,464 persons to become ill. A protozoal parasite (Giardia lamblia or Cryptosporidium) was identified as the etiologic agent for seven of the 11 outbreaks for which an agent was determined. Five (71%) of the outbreaks caused by protozoa were associated with a surface-influenced groundwater source. One outbreak of cryptosporidiosis was associated with filtered and chlorinated surface water. Shigella sonnei and hepatitis A virus were implicated in one outbreak each; both were linked to consumption of contaminated well water. Two outbreaks due to acute chemical poisoning were reported; one had an associated fatality. No etiology was established for 23 (68%) of the 34 outbreaks, including the largest one reported during this period, in which an estimated 9,847 persons using a filtered surface water supply developed gastroenteritis. Most (76%) of the 34 outbreaks were associated with a well water source. Twenty-one states reported 39 outbreaks associated with recreational water, in which an estimated 1,825 persons became ill. The most frequently reported illness was hot tub- or whirlpool-associated Pseudomonas dermatitis (12 outbreaks). Of 11 outbreaks of swimming-associated gastroenteritis, six were caused by Giardia or Cryptosporidium, including three outbreaks associated with chlorinated, filtered pool water. The first reported outbreak of Escherichia coli O157:H7 infection associated with recreational exposure occurred during this period. Primary amebic meningoencephalitis, caused by Naegleria fowleri infection, resulted in six deaths. INTERPRETATION: The number of waterborne disease outbreaks reported per year has not changed substantially in the past 5 years. However, etiologic agents only recently associated with waterborne disease, such as E. coli O157:H7 and Cryptosporidium, are being reported more frequently and from new settings. Water quality data for outbreaks during the period 1991-1992 indicate that available water disinfection technology is not always in place or used reliably.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8232180 TI - Silicosis surveillance--Michigan, New Jersey, Ohio, and Wisconsin, 1987-1990. AB - PROBLEM/CONDITION: Improved surveillance for silicosis is needed to target interventions to prevent this occupational lung disease caused by the inhalation of crystalline silica dust. REPORTING PERIOD COVERED: 1987-1990. DESCRIPTION OF SYSTEMS: State-based silicosis surveillance and intervention programs have been developed in Michigan, New Jersey, Ohio, and Wisconsin as part of the Sentinel Event Notification System for Occupational Risks (SENSOR) Program, initiated in 1987 by the National Institute for Occupational Safety and Health (NIOSH). RESULTS: From 1987 through 1990, the SENSOR program confirmed a total of 430 cases of silicosis reported from these four states. Overall, approximately 60% of these cases were in workers employed in primary metal industries, although the types of industries in which cases occurred varied by state. Some cases were attributable to relatively recent exposure, including new cases in seven persons first exposed since 1980 in New Jersey. Silicosis case reports have prompted measurement of respirable silica concentrations at 25 Michigan work sites, and 14 (56%) of these sites were found to have levels that exceeded the legally permissible exposure level. INTERPRETATION: The silicosis surveillance and intervention strategies piloted by state health departments in the NIOSH-funded SENSOR Program have demonstrated the feasibility and effectiveness of identifying specific silica-using work sites that need preventive intervention. ACTIONS TAKEN: On the basis of initial experience in these four states, NIOSH developed guidelines for state-based silicosis surveillance and awarded SENSOR cooperative agreements to three additional states where the applicability of these surveillance methods will be further evaluated. PMID- 8232181 TI - Sensitivity of multiple-cause mortality data for surveillance of deaths associated with head or neck injuries. AB - PROBLEM/CONDITION: Multiple-cause mortality data was assessed as a source of information for surveillance of deaths associated with head or neck injuries. REPORTING PERIOD COVERED: 1985-1986 DESCRIPTION OF SYSTEM: Data on causes of death were abstracted from death certificates in New Mexico and coded according to criteria of the International Classification of Diseases, Ninth Revision (ICD 9). Deaths with an external cause-of-death (E) code as the underlying cause of death and one or more head or neck injury nature-of-condition (N) codes as contributing causes of death were considered head or neck injury deaths. These data were compared with data for head or neck injury deaths obtained from computerized records from the New Mexico Office of the Medical Investigator (OMI). Data for alcohol or drug use were abstracted from both systems. RESULTS: Of the 699 head or neck injury deaths coded by the New Mexico OMI system in 1985 1986, 536 were identified as head or neck injury deaths in multiple-cause mortality data (sensitivity = 76.7%). Firearms were the leading cause of head or neck injury deaths, followed by motor vehicles. Multiple-cause mortality data contained alcohol codes for only 3.7% of OMI records with blood alcohol concentrations > or = 0.10 mg/dL and contained drug codes for none of the OMI records with positive toxicology tests for drugs. INTERPRETATION: The sensitivity of multiple-cause mortality data was relatively high for surveillance of head and neck injury deaths.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232182 TI - [Glutamine (Gln) supplementation in septic rats]. AB - The objectives of these experiments were to investigate the effect of Gln supplementation on protein metabolism and immune function in septic rats. (Experiment 1): 73 SD female rats were catheterized for TPN into the jugular vein on day 1. On day 4, the rats were randomized into 4 group: 1) control (C)+Standard TPN (STPN), 2) (C)+Gln TPN (GTPN), 3) sepsis (S)+STPN, 4) S+GTPN. Sepsis was induced by injection of 10(10) C. Coli/kg from the TPN catheter. U-14C leucine or 15N2-Urea was given before sacrifice on day 5. (Experiment 2): 48 SD male rats were randomized into 3 groups, 1) normal control rat (NC), fed as lib. 2) peritonitis (P)+STPN, 3) P+GTPN. On day 1, 34 rats were catheterized and either STPN or GTPN was begun. On day 3, 6 hours after serum cecum ligation and puncture, resuscitation was done. On day 5, rats were sacrificed. The results were as follows: 1) FSR of ileum, proximal colon, distal colon and muscle were augmented by GTPN, 2) Sepsis caused a significant increase of urea production, but GTPN prevented this increase, 3) lymphocyte blastogenation was decreased with sepsis, but GTPN improved this reduction, 4) Phagocytic index was higher with GTPN than STPN. We concluded that Gln supplementation would prevent from leading the patients with severe infection to the multiple organ failure. PMID- 8232183 TI - [The effect of free radicals from non-parenchymal cells (NPC) of the liver on the development of liver metastases in rat]. AB - Tumor cells (AH130 hepatoma cell originated from rat) were injected intraportally into Donryu rats to produce liver metastases 21 days later. Phagocyte cells activity was depressed by the administration of Silica, which significantly increased the number of surface liver metastases. Phagocyte cells were stimulated by beta 1-3-glucan, which significantly reduced the number of metastases. And the administration of free radical scavenger (SOD, Catalase) increased the number of metastases. Non parenchymal cells (NPC) of the liver play a main role of self defence line for portally liver metastases. Then free radical from these cells were noticed in this study. NPC were isolated, from pronase perfused rat liver. O2- production by activated NPC was measured by chemiluminescence with CLA. NPC activated by beta 1-3-glucan added sera increased the luminescence of CLA, and SOD depressed the production of chemiluminescence. SOD activity of hepatocytes and tumor cells (AH130) were measured by NBT methods. Hepatocytes had high potential production of SOD, in contrast AH130 had poor production. These results suggest that free radicals from liver NPC was important for protecting liver metastases. PMID- 8232184 TI - [Regional lymph node recurrence in patients with papillary thyroid carcinoma who did not undergo neck dissection]. AB - It is important and difficult problem to manage patients with thyroid nodules which are indeterminate as papillary thyroid carcinoma (PTC) before surgery and identified as PTC after surgery. This paper highlighted whether immediate reoperation for such patients should be undergone or not. The prognosis of 196 patients with PTC who underwent lobectomy or subtotal thyroidectomy without neck dissection were studied (non-dissection group). The controls consisted of 783 patients with non-advanced PTC who underwent radical surgery during the same period (dissection group). There was no death due to PTC in both groups. Distant metastasis was observed 1.5% in non-dissection group and 2.2% in the dissection group. Local recurrence was observed 9.7% in the non-dissection group and 11.2% in the dissection group. The results suggested that immediate reoperation for the patient's who did not undergo neck dissection, is not necessary. It is thought that all of lymph node metastasis detected only by histology does not always behave in same. PMID- 8232185 TI - [Study of the localization and the concentration of superoxide dismutase in various thyroid disorders]. AB - The localization of Cu/Zn and Mn superoxide dismutase (SOD), which catalyzes the dismutation of superoxide radicals (O2-) to O2 and H2O2, in various thyroid disorders was studied by an immunohistochemical technique in 20% formalin fixed paraffin embedded thin sections using anti-human Cu/Zn and Mn-SOD antibodies. The concentrations of both SODs in those tissues were also measured by a sandwich enzyme immunoassay technique. Cu/Zn SOD was immunocytochemically stained in most of papillary carcinoma, and some of other disorders. In the normal follicular cells it localized in the perinuclear cytoplasm, whereas in the tumor or hyperplastic cells it was in the cytoplasm homogeneously. Mn SOD was strongly stained in papillary carcinoma and papillary growing cells in adenoma and Graves' disease. The concentrations of Cu/Zn and Mn SOD in every thyroid disorder were higher than those in normal thyroid, and it was much higher in the malignant tumors. The ratio of the concentration of Mn SOD against Cu/Zn SOD in the tissues was higher only in papillary carcinoma compared with that in normal thyroid. In conclusion, SOD seems to be relating to the cell proliferation and differentiation in the thyroid since both SODs increased in tumors and hyperplastic disorders, and Cu/Zn SOD changed its localization and Mn SOD existed predominantly in papillary carcinoma or papillary growing cells. PMID- 8232186 TI - [New surgical strategy for secondary hyperparathyroidism]. AB - Extopic parathyroid glands and rudimentary glands which are not detected during initial parathyroidectomy may develop hyperplasia in the future, causing recurrence of hyperthyroidism. We have devised an improved surgical strategy to prevent the recurrence of hyperparathyroidism. The strategy is to dissect all central lymph nodes and fatty tissue according to the procedure of the dissection of thyroid cancer. The upper level of the dissection area is the upper edge of the thyroid cartilage and the lateral level of the dissection includes the lateral edge of the bilateral carotid sheath. Bilateral transcervical thymectomy is always included in this strategy. Upon confirming that more than 4 glands have been removed by intraoperative stamp cytology, total parathyroidectomy is terminated and 80 mg of glands is autotransplanted to the muscle of one forearm. We have performed this operation on 30 patients since 1987. More than 4 glands could be extirpated in every case, and supernumerary glands were detected in 6 cases (7 glands in 1 case and 5 glands in 4 cases). We have had no operative side effects in this series of 30 patients, and we have not experienced any persistent or recurrent hyperparathyroidism after any of these operations. PMID- 8232187 TI - [Time-dependent interrelationships between pathological prognostic factors and relapse rate in breast cancer patients]. AB - Eleven pathological factors among 462 patients with primary breast carcinoma (1960-1968) were analysed paying a special attention to changes of the influence on relapse. Factors were included; tumor size (T), extent of nodal involvement (N), nuclear grade (NG), fat infiltration (F), tumor infiltrating lymphocyte (TIL), tumor necrosis (NEC), lymphatic & vessel permeation (LY) and others. Cox regression analysis on 3 non-overlapping intervals (within 2 years, 2-5 years and after 5 years) using the seven factors selected by Cox analysis in the whole periods under the condition of p < 0.10 showed that significant factors were N, NG and T within 2 years, N, TIL, NG and NEC in 2-5 years, F, N, TIL and T after 5 years, respectively (p < 0.05), in the following order. TIL means the relapse suppressing factor and others mean relapse factors. As for the changes of beta value, it decreased with time for N and NG, but contrarily increased with time for F. This means that N and NG participate mainly in an early relapse and F mainly in a delayed relapse. TIL showed a significant relapse suppressing factor only after 2 years. These results revealed that prognostic factors have its altered influence on relapse with time, namely a distinct time-dependency. PMID- 8232188 TI - [Clinical experience of VA-bypass as a cardiac assist system using centrifugal pump with membranous oxygenator in patients with profound ventricular failure after cardiac surgery]. AB - Twelve patients including 6 ischemic heart diseases, 5 valvular diseases, and one adult atrial septal defect combined with mitral and tricuspid valve insufficiency were treated with circulatory assist system for postoperative severe low-cardiac output syndrome. Intra-aortic balloon pumping (IABP) was used in all cases. In early 7 cases, left heart bypass system was employed including one roller pump, one left ventricular assist device (LVAD), and 5 centrifugal pump. Only one LVAD case could be weaned but died of sustained ventricular arrhythmias 5 days postoperatively. In recent 5 cases, veno-arterial bypass (VA-bypass) was employed using centrifugal pump with membranous oxygenator. Venous blood was drained from the right atrium and oxygenated blood was returned to the right axillary artery. Real time reinfusion of drained blood from intrapericardial and mediastinal tube to the central vein served efficiently to maintain hemodynamics and to save homologous blood transfusion. Three of 5 cases who were treated with Va-bypass survived and are doing well. We conclude that VA-bypass consisting with centrifugal pump and oxygenator returning oxygenated blood to the right axillary artery combined with IABP were useful strategies as biventricular support for profound cardiac failure following surgery. PMID- 8232189 TI - [Case of diffuse esophageal spasm treated by long myotomy]. AB - Diffuse esophageal spasm is a rare condition, which has generally been treated conservatively in Japan. A case of this disorder treated successfully by long myotomy is reported with a knowledge of prolonged pressure monitoring of the esophagus. A 56-year-old woman was admitted to our hospital with dysphagia. Barium swallow and esophagoscopy showed contraction of the esophagus, and manometry showed normal peristalsis, so the diagnosis of achalasia was ruled out. Prolonged pressure monitoring of the esophagus showed spastic contractions with the pressure over 130 mmHg that continued for 25 sec during meals, which led us to the diagnosis of diffuse esophageal spasm. Extramucosal long myotomy of the esophagus and a modified Belsey Mark IV operation were performed. The postoperative course was satisfactory and esophageal functional tests showed no spasms. The patient regained weight without dysphagia. PMID- 8232190 TI - [A new model of liver metastasis of gastric cancer in nude mice with orthotopic transplantation (injection of tumor into gastric wall): preliminary report]. PMID- 8232191 TI - [Study of the serum interleukin-2 with endotoxemia and fungemia patients: preliminary report]. PMID- 8232192 TI - [Expression of a novel cancer antigen coagulant cancer antigen 1 (CCA-1) on various human cancers: preliminary report]. PMID- 8232193 TI - The membrane topology of the Rhizobium meliloti C4-dicarboxylate permease (DctA) as derived from protein fusions with Escherichia coli K12 alkaline phosphatase (PhoA) and beta-galactosidase (LacZ). AB - The Rhizobium meliloti dctA gene encodes the C4-dicarboxylate permease which mediates uptake of C4-dicarboxylates, both in free-living and symbiotic cells. Based on the hydrophobicity of the DctA protein, 12 putative membrane spanning regions were predicted. The membrane topology was further analysed by isolating in vivo fusions of DctA to Escherichia coli alkaline phosphatase (PhoA) and E. coli beta-galactosidase (LacZ). Of 10 different fusions 7 indicated a periplasmic and 3 a cytoplasmic location of the corresponding region of the DctA protein. From these data a two-dimensional model of DctA was constructed which comprised twelve transmembrane alpha-helices with the amino-terminus and the carboxy terminus located in the cytoplasm. In addition, four conserved amino acid motifs present in many eukaryotic and prokaryotic transport proteins were observed. PMID- 8232194 TI - Maternal inheritance of P cytotype in Drosophila melanogaster: a "pre-P cytotype" is strictly extra-chromosomally transmitted. AB - In Drosophila melanogaster, transposition of the P element is under the control of a cellular state known as cytotype. The P cytotype represses P transposition whereas the M cytotype is permissive for transposition. In the long-term, the P cytotype is determined by chromosomal P elements but over a small number of generations it is maternally inherited. In order to analyse the nature of this maternal inheritance, we tested whether a maternal component can be transmitted without chromosomal P elements. We used a stable determinant of P cytotype, linked to the presence of two P elements at the tip of the X chromosome (1A site) in a genome devoid of other P elements. We measured P repression capacity using two different assays: gonadal dysgenic sterility (GD) and P-lacZ transgene repression. We show that zygotes derived from a P cytotype female (heterozygous for P (1A)/balancer devoid of P copies) and which inherit no chromosomal P elements from the mother, have, however, maternally received a P-type extra chromosomal component: this component is insufficient to specify the P cytotype if the zygote formed does not carry chromosomal P elements but can promote P cytotype determination if regulatory P elements have been introduced paternally. We refer to this strictly extra-chromosomally inherited state as the "pre-P cytotype". In addition, we show that a zygote that has the pre-P cytotype but which has not inherited any chromosomal P elements, does not transmit the pre-P cytotype to the following generation. The nature of the molecular determinants of the pre-P cytotype is discussed. PMID- 8232195 TI - Molecular analysis of allelic polymorphism at the AAT2 locus of alfalfa. AB - Aspartate aminotransferase (AAT) plays a key enzymatic role in the assimilation of symbiotically fixed nitrogen in legume root nodules. In alfalfa, two distinct genetic loci encode dimeric AAT enzymes: AAT1, which predominates in roots, and AAT2, which is expressed at high levels in nodules. Three allozymes of AAT2 (AAT2a, -2b and -2c), differing in net charge, result from the expression of two alleles, AAT2A and AAT2C, at this locus. Utilizing antiserum to alfalfa AAT2, we have previously isolated from an expression library one AAT2 cDNA clone. This clone was used as a hybridization probe to screen cDNA libraries for additional AAT2 cDNAs. Four different clones were obtained, two each that encode the AAT2a and AAT2c enzyme subunits. These two sets of cDNAs encode polypeptides that differ in net charge depending upon the amino acid at position 296 (valine or glutamic acid). Within each set of alleles, the two members differ from each other by the presence or absence of a 30 bp (ten amino acid) sequence. The presence or absence of this ten amino acid sequence has no effect on the size or charge of the mature AAT2 protein because it is located within the region encoding the protein's transit peptide, which is proteolytically removed upon transport into plastids. The data suggest that a deletion event has occurred independently in two AAT2 progenitor alleles, resulting in the four allelic cDNA variants observed. The deletion of this ten amino acid sequence does not appear to impair the normal maturation of the enzyme. PMID- 8232196 TI - Genetic organization of the streptokinase region of the Streptococcus equisimilis H46A chromosome. AB - The complete nucleotide sequences of four genes and one open reading frame (ORF1) adjacent to the streptokinase gene, skc, from Streptococcus equisimilis H46A were determined. These genes are encoded on the opposite DNA strand to skc and are arranged as follows: dexB-abc-lrp-skc-ORF1-rel. The dexB gene, coding for an alpha-glucosidase (M(r) 61,733), and abc, encoding an ABC transporter (M(r) 42,080), are similar to the dexB and msmK genes, respectively, from the multiple sugar metabolism operon of S. mutans. The lrp gene specifies a leucine-rich protein (M(r) 32,302) that has a leucine-zipper motif at its C-terminus. The function of the Lrp protein is not known but appeared to be detrimental when overexpressed in Escherichia coli. Although lrp appears not to be an essential gene, as judged by plasmid insertion mutagenesis, it is conserved in all streptococcal strains carrying a streptokinase gene. The rel gene showed significant homology to the E. coli relA and spoT genes involved in the stringent response to amino acid deprivation. Multiple alignment of the amino acid sequences of Rel (M(r) 83,913), RelA and SpoT revealed 59.4% homology of the primary structures. Northern hybridization analyses of the genes in the skc region showed skc to be transcribed most abundantly. In addition to transcripts for skc, monocistronic mRNAs were detected for all three genes divergently transcribed from skc. Although there was also some read-through transcription from lrp into abc, and from abc into dexB, the transcription pattern suggests a high degree of transcriptional and functional independence not only of skc but also abc and dexB. Prominent structural features in intergenic regions included a static DNA bending locus located upstream and a putative bidirectional transcription terminator downstream of skc. PMID- 8232197 TI - Differential expression of the partially duplicated chloroplast S10 ribosomal protein operon. AB - The chloroplast S10 ribosomal protein operon is partially duplicated in many plants because it initiates within the inverted repeat of the circular chloroplast genome. In spinach, the complete S10 operon (S10B) spans the junction between inverted repeat B (IRB) and the large single-copy (LSC) region. The S10 operon is partially duplicated in the inverted repeat A (IRA), but the sequence of S10A completely diverges from S10B at the junction of S10A and the LSC region. The DNA sequence shared by S10A and S10B includes trnI1, the rpl23 pseudogene (rpl23 psi), the intron-containing rpl2 and rps19, which is truncated in S10A at the S10A/LSC junction (rps19'). Transcription of rps19' from the promoter region of S10A could result in the synthesis of a mutant S19 protein. Analysis of RNA accumulation and run-on transcription from S10A and S10B using unique probes from the S10A/LSC and S10B/LSC junctions reveals that expression of S10A is reduced. The difference in S10A and S10B expression appears to be the result of reduced transcription from S10A, rather than differences in RNA stability. Transcription of S10B can initiate at three distinct promoter regions, P1, P2 and P3, which map closely to transcripts detected by S1 nuclease analysis. P1 is located upstream of trnI1 and has the highest transcription initiation frequency in vitro of the three promoter regions. The DNA sequence of P1 is most similar to the chloroplast promoter consensus DNA sequence. Interference by the highly and convergently transcribed psbA-trnH1 operon is considered as a mechanism to explain the reduced activity of the S10A promoters. PMID- 8232198 TI - Functional cis-element sequence requirements for suppression of gene expression by the TNPA protein of the Zea mays transposon En/Spm. AB - TNPA, one of the two transposition proteins encoded by the En/Spm transposable elements of Zea mays, suppresses the expression of genes that contain an appropriate cis element. Suppression can be monitored in tobacco protoplasts in a transient expression assay as follows. The plant promoter-driven expression of the Escherichia coli-glucuronidase (GUS)-encoding gene, uidA, is repressed in the presence of TNPA if the GUS gene contains a functional cis element in the untranslated RNA leader sequence. Earlier, we found that the minimal cis element is composed of two 12 bp sequences in a tail-to-tail inverted orientation. Each 12 bp sequence is sufficient to bind TNPA in vitro and can be thought of as a half-site in the cis element. Here, we investigated the sequence requirements of the minimal cis element. Our observations support our expectations that a functional cis element must provide a template to which two TNPA molecules can bind in the correct orientation. Sequences within the half-sites can be altered as long as the eight bases that make up the consensus binding sites are not changed. However, we found the following unexpected sequence specificities. Firstly, some changes to the consensus binding sequence can be tolerated in one half-site, as long as the other site matches the consensus. Secondly, although the region between the half-sites can vary in sequence and in length between two and four bases, a thymidine residue is not tolerated directly 5' preceding the second half-site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232199 TI - Antiparallel expression of the sense and antisense transcripts of maize alpha tubulin genes. AB - In all eukaryotes alpha- and beta-tubulins are encoded by small families of closely related genes and are highly conserved. In Zea mays, at least six different alpha-tubulin coding sequences are known. We describe the isolation from scutellar nodes of the maize inbred line W22 of a clone (CTM5) coding for an alpha-tubulin. On the basis of the 3' end nucleotide sequence, this clone can be assigned to the already reported tua4 gene. Northern analysis demonstrates that CTM5 encodes a 1.5 kb transcript, which is expressed in different tissues of the seed and of the seedling. In order to define the spatial and temporal expression of alpha-tubulin genes, in situ hybridization experiments were performed on these tissues. Unexpectedly, a specific signal was detected with both antisense and sense RNA strands. Temporal and spatial distribution of the two RNAs, however, shows that high levels of the two transcripts are always discordant. In tissues where sense transcripts are highly abundant (embryos at various developmental stages, root tips, pollen grains), the antisense transcripts are expressed in relatively small amounts, while in pericarp, coleoptile, leaves, and scutellar node, where antisense transcripts accumulate, the sense transcript only reaches a very low level. Northern analysis using single-stranded DNA probes confirmed the presence of an antisense transcript of 1.5 kb, prompting speculation about the role of this transcript in the regulation of the expression of alpha-tubulin genes. PMID- 8232200 TI - Identification and characterization of stylar glycoproteins associated with self incompatibility genes of Japanese pear, Pyrus serotina Rehd. AB - Japanese pear (Pyrus serotina Rehd.) exhibits gametophytic self-incompatibility. Following our previous findings that basic ribonucleases in the styles of Japanese pear are associated with self-incompatibility genes (S-RNases), stylar proteins with high pI values were analyzed by two-dimensional gel electrophoresis further to characterize S-RNases. A group of basic proteins of about 30 kDa associated with self-incompatibility genes were identified. These proteins contained sugar chains which reacted with concanavalin A and wheat germ agglutinin, and thus were designated as S-glycoproteins of Japanese pear. The fact that the S-glycoprotein was expressed at a much lower level in a self compatible mutant than in the original variety suggested a role of S glycoproteins in mediating self-incompatibility of Japanese pear. Immunoblot analysis indicated that S-glycoproteins are identical to previously identified S RNases. The S-glycoproteins were predominantly expressed in the style, in the ovary in trace amounts, and not in leaf, pollen or germinated pollen. The N terminal amino acid sequences of the S-glycoproteins showed homology not only with each other but also with those of the S-allele-associated proteins from plants of the family Solanaceae at levels of about 30-50%. PMID- 8232201 TI - Cloning sequencing and expression of the gene for cytochrome P450meg, the steroid 15 beta-monooxygenase from Bacillus megaterium ATCC 13368. AB - A 4.3 kb EcoRI fragment carrying the gene for cytochrome P450meg, the steroid-15 beta-monooxygenase from Bacillus megaterium ATCC 13368, was cloned and completely sequenced. The gene codes for a protein of 410 amino acids and was expressed in Escherichia coli and B. subtilis. Protein extracts from the recombinant E. coli strains were able to hydroxylate corticosteroids in the 15 beta position when supplemented with an extract from a P450- mutant of B. megaterium ATCC 13368 as a source of megaredoxin and megaredoxin reductase. In contrast, 15 beta hydroxylation was obtained in vitro and in vivo without the addition of external electron transfer proteins, when cytochrome P450meg was produced in B. subtilis 168. Protein extracts from nonrecombinant B. subtilis 168 could also support the in vitro hydroxylation by cytochrome P450meg produced in E. coli. PMID- 8232202 TI - Activity of the yeast MAP kinase homologue Slt2 is critically required for cell integrity at 37 degrees C. AB - Deletion of the SLT2 gene of Saccharomyces cerevisiae, which codes for a homologue of MAP (mitogen-activated) protein kinases, causes an autolytic lethal phenotype in cells grown at 37 degrees C. The gene encodes domains characteristic of protein kinases, which include a lysine (at position 54) that lies 19 residues from a glycine-rich cluster, considered to be the putative ATP binding site. The ability of three mutant alleles of SLT2 generated by site-directed mutagenesis, namely E54 (glutamic acid), R54 (arginine) and F54 (phenylalanine), to complement slt2 mutants was tested. All three failed to complement the autolytic phenotype and were unable to restore growth and viability of cells. A strain obtained by transplacement of slt2-F54 also behaved as a thermosensitive autolytic mutant. By immunoprecipitation with polyclonal antibodies raised against Slt2 protein expressed in Escherichia coli, it was possible to confirm that alteration of the lysine-54 residue did not affect the stability of the protein, thus allowing us to conclude that activity of the Slt2 protein kinase is critically required for growth and morphogenesis of S. cerevisiae at 37 degrees C. A significant fraction of the mutant cell population lysed at 24 degrees C and the cells displayed a characteristic alteration of the surface consisting of a typical depression in an area of the cell wall. At 37 degrees C, the cell surface was clearly disorganized. PMID- 8232203 TI - Characterization of the Staphylococcus aureus chromosomal gene pcrA, identified by mutations affecting plasmid pT181 replication. AB - The Staphylococcus aureus chromosomal gene pcrA, identified by mutations, such as pcrA3, that affect plasmid pT181 replication, has been cloned and sequenced. The pcrA gene encodes a protein with significant similarity (40% identity) to two Escherichia coli helicases: the helicase II encoded by the uvrD gene and the Rep helicase. The pcrA3 mutation was found to be a C to T transition leading to a threonine to isoleucine substitution at amino acid residue 61 of the protein. The pcrA gene seems to belong to an operon containing at least one other gene, tentatively named pcrB, upstream from pcrA. The PcrA protein was shown to be essential for cell viability and overproduction has deleterious effects on the host and plasmid replication. PMID- 8232204 TI - Nucleotide sequence analysis of five putative Streptomyces griseus genes, one of which complements an early function in daunorubicin biosynthesis that is linked to a putative gene cluster involved in TDP-daunosamine formation. AB - Sequence analysis of the lkmB region of the daunorubicin biosynthetic gene cluster of Streptomyces griseus JA3933 revealed two contiguous open reading frames (ORF) in the same orientation, and three ORFs in the opposite orientation together extending over a 4.6 kb region adjacent to a homologue of the S. peucetius dnrJ gene. ORF1 complemented in trans the lkmB mutation, which seems to affect an early step in daunorubicin biosynthesis. Its deduced product showed no similarity to any known enzyme in the databases. The mutation in ORF1 was localised to a C-T transition at position 1172, leading to the change from a glycine to aspartic acid in the deduced protein. The lack of any homology to known polyketide synthesis enzymes indicates a regulatory role for the product of ORF1, despite the ability of lkmB mutants to further metabolise alkanoic acid. The genes of the oppositely oriented cluster seem to be involved in sugar metabolism. The putative ORF3 protein revealed strong homology to eukaryotic acyl CoA dehydrogenases and might encode an enzyme for the oxidoreduction preceding the introduction of the amino group into daunosamine, and the ORF4 protein is homologous to several epimerases, central enzymes in the formation of the L-2,3,6 trideoxy-3-aminohexoses from TDP-D-glucose. ORF5 seems also to be related to enzymes metabolising nucleotide-activated hexoses. PMID- 8232205 TI - Functional expression of the tellurite resistance determinant from the IncHI-2 plasmid pMER610. AB - The transpositional phage MudI 1734 lacZ was used to construct transcriptional fusions within the plasmid pMJ611, which contains the cloned tellurite resistance (TeR) determinant of the IncHI-2 plasmid pMER610. A series of 70 MudI insertions, in both orientations, causing loss of tellurite resistance in pMJ611, mapped within a 4.3 kb region which included the genes terA-terD and a 0.4 kb region upstream of the site previously reported as the 5' limit of the TeR determinant. Expression of beta-galactosidase from these transcriptional fusions, including those involving the 5' upstream region, occurred only from inserts transcribed in the direction terA-terD, confirming the transcriptional orientation of the TeR determinant deduced from DNA sequence analysis. Sixteen of the tellurite sensitive MudI fusions, distributed over the entire determinant and in both orientations, showed the same pattern of expression when transferred by conjugation and homologous recombination to pMER610, except that the beta galactosidase levels were consistently 2- to 3-fold higher in the parent plasmid. Northern analysis with a DNA probe spanning the TeR determinant identified five transcripts of 4.8, 4.0, 2.7, 1.5 and 1.0 kb synthesised by pMER610. Further hybridisations with DNA probes defining sub-sections of the TeR determinant, together with DNA sequence analysis, suggested the presence of three transcriptional start sites, at approximately 0.9 and 0.1 kb upstream of terA, and near the junction between terC and terD. Three transcriptional termination sites, located within terA, near the terC-terD junction and at the 3' end of terE are also indicated. Both the expression of beta-galactosidase from the MudI fusions and the synthesis of ter gene transcripts are constitutive and were not affected by prior exposure of cultures to sub-toxic levels of tellurite. Further DNA sequence analysis reveals that the extensive homology between terD and terE extends to a section of terA. PMID- 8232206 TI - Transcription of the yeast mitochondrial genome requires cyclic AMP. AB - Using various mutant strains and nutritional manipulations, we investigated a potential role for cyclic AMP (cAMP) in the regulation of mitochondrial (mt) gene expression in the yeast Saccharomyces cerevisiae. In RAS mutants known to have either abnormally low or high cellular levels of this nucleotide, we show that both mt transcription rate and overall mt transcript levels vary directly with cellular cAMP levels. We further show that nutritional downshift of actively growing cells causes a severe, rapid fall in cAMP levels, and that this fall is concomitant with the stringent mt transcriptional curtailment that we and others have previously shown to follow this nutritional manipulation. In in vitro mt transcription assays using intact organelles from downshifted and actively growing cells, stringently curtailed mt gene expression can be restored to 75% of control levels by addition of cAMP to the assay mix. Consistent with these observations a RAS2vall9 mutant strain, which cannot adjust cAMP levels in response to external stimuli, shows no mt stringent response following nutritional downshift. We also demonstrate a significant but transient increase in both mt transcript levels and mt transcription rate following shift of actively respiring wild-type cells to glucose-based medium, a manipulation known to cause a short-lived pulse of cAMP in yeast; similar manipulation of the RAS2vall9 mutant strain generates no such response. Taken together all these observations indicate that cellular cAMP levels are involved in the regulation of mt transcription in yeast. Moreover, the lack of a mt stringent transcriptional response following downshift in a strain in which the BCY1 gene had been insertionally inactivated suggests that cAMP may influence mt transcription via a mt cAMP-dependent protein kinase. These results link mt gene expression with mechanisms governing growth control and nutrient adaptation in yeast, and they provide a means by which mt gene expression might be coordinated with that of related nuclear genes. PMID- 8232207 TI - pMH2, a small plasmid bearing the nif gene cluster of Enterobacter agglomerans 333 as an excisable cassette. AB - A small plasmid containing the entire nif gene cluster of Enterobacter agglomerans 333 as an excisable cassette has been constructed, using pACYC177 as a vector. Two cosmid clones taken from a gene library of E. agglomerans plasmid pEA3 were used as a source of nif genes. A SmaI fragment of peaMS2-2, containing the H,D,K,Y,E,N,X,U,S,V,W,Z,M,L,A and B genes and an ApaI fragment of peaMS2-16 containing nif A,B,Q,F and J were selected to construct pMH2. The resulting plasmid of 33 kb carries the complete nif gene cluster as a nif cassette on a single XbaI fragment. The nif construct pMH2 in Escherichia coli strains has significant nitrogenase activity compared to wild-type E. agglomerans 333. The nif gene cluster construct was found to be very stable. PMID- 8232208 TI - Expression of a Drosophila melanogaster amber suppressor tRNA(Ser) in Caenorhabditis elegans. AB - The purpose of this study was to test a cloned amber-suppressing tRNA(Ser) gene derived from Drosophila melanogaster for its ability to produce amber suppression in the nematode Caenorhabditis elegans. To date, all characterized nonsense suppressors in C. elegans have been derived from tRNA(Trp) genes. Suppression was assayed by monitoring the reversal of a mutant tra-3 phenotype among individuals transformed with the cloned Drosophila suppressor gene. An amber allele of tra-3 results in masculinization of XX animals with accompanying sterility. Complete suppression was observed among the transformants. The presence of the heterologous transgene, in both suppressed experimental animals and controls injected with a non-suppressing wild-type Drosophila tRNA(Ser) gene, was verified by PCR amplification of DNA from single worms using primers flanking the tRNA(Ser) gene. Suppression by the heterologous transgene was comparable in quality to that produced by endogenous C. elegans suppressors, and, in frequency as well as quality, to that produced by a transgenic C. elegans tRNA(Trp)-derived suppressors. Thus, a heterologous suppressor gene will function in C. elegans, and it need not be based on tRNA(Trp). PMID- 8232209 TI - Cloning and characterization of the scrA gene encoding the sucrose-specific Enzyme II of the phosphotransferase system from Staphylococcus xylosus. AB - By insertional mutagenesis with the staphylococcal transposon Tn551, mutants of Staphylococcus xylosus were isolated that were unable to utilize sucrose. One of these was found to be deficient in sucrose uptake. The genomic region containing this sucrose uptake gene of Staphylococcus xylosus (scrA) was cloned in Staphylococcus carnosus. The scrA gene was further localized to a 4.4 kb DNA fragment by complementation of the sucrose transport-deficient S. xylosus mutant. The DNA sequence analysis of the scrA region revealed three open reading frames, one of which encodes a protein of 480 amino acids (51.335 kDa) with significant similarity to sucrose-specific Enzymes II of phosphoenolpyruvate-dependent carbohydrate phosphotransferase systems (PTS). A protein with an apparent molecular weight of 50 kDa was obtained in Escherichia coli by expression of scrA with the bacteriophage T7 RNA polymerase promoter system. Transcriptional start sites of the scrA gene were localized by primer extension analysis to positions 46 and 49 nucleotides upstream of the scrA start codon. No additional sucrose utilization genes are encoded close to scrA on the S. xylosus chromosome. PMID- 8232210 TI - The vestigial locus of Drosophila melanogaster is involved in resistance to inhibitors of dTMP synthesis. AB - The vestigal (vg) gene encodes a nuclear protein which plays a major role in the formation of the wing of Drosophila. Resistance or sensitivity to aminopterin, an inhibitor of the dihydrofolate reductase enzyme in D. melanogaster, seems to be associated with a specific alteration in vg gene function. Wild-type and vg mutant strains selected for growth on increasing concentrations of aminopterin display changes in physiological and biochemical parameters such as viability on normal and aminopterin-containing media, duration of development, wing phenotype, dihydrofolate reductase activity, and cross-resistance to fluorodeoxyuridine (FUdR) and to methotrexate. Our results indicate that the mechanisms of resistance differ in the wild-type and mutant strains. The vg83b27 mutant, in which the major part of intron 2 of the vg gene is deleted, is associated with a high rate of resistance to FUdR, an inhibitor of thymidylate synthetase. Moreover, vg83b27/vgBG heterozygotes, which are wild type when grown on normal medium, display a strong vg phenotype when grown on aminopterin. Our results indicate a role for the vestigial locus in mediating resistance to inhibitors of dTMP synthesis. PMID- 8232211 TI - Kanamycin resistance as a selectable marker for plastid transformation in tobacco. AB - We report on a novel chimeric gene that confers kanamycin resistance on tobacco plastids. The kan gene from the bacterial transposon Tn5, encoding neomycin phosphotransferase (NPTII), was placed under control of plastid expression signals and cloned between rbcL and ORF512 plastid gene sequences to target the insertion of the chimeric gene into the plastid genome. Transforming plasmid pTNH32 DNA was introduced into tobacco leaves by the biolistic procedure, and plastid transformants were selected by their resistance to 50 micrograms/ml of kanamycin monosulfate. The regenerated plants uniformly transmitted the transplastome to the maternal progeny. Resistant clones resulting from incorporation of the chimeric gene into the nuclear genome were also obtained. However, most of these could be eliminated by screening for resistance to high levels of kanamycin (500 micrograms/ml). Incorporation of kan into the plastid genome led to its amplification to a high copy number, about 10,000 per leaf cell, and accumulation of NPTII to about 1% of total cellular protein. PMID- 8232212 TI - Enhanced detection of polymorphic DNA by multiple arbitrary amplicon profiling of endonuclease-digested DNA: identification of markers tightly linked to the supernodulation locus in soybean. AB - Multiple endonuclease digestion of template DNA or amplification products can increase significantly the detection of polymorphic DNA in fingerprints generated by multiple arbitrary amplicon profiling (MAAP). This coupling of endonuclease cleavage and amplification of arbitrary stretches of DNA, directed by short oligonucleotide primers, readily allowed distinction of closely related fungal and bacterial isolates and plant cultivars. MAAP analysis of cleaved template DNA enabled the identification of molecular markers linked to a developmental locus of soybean (Glycine max L. Merrill). Ethyl methane sulfonate (EMS)-induced supernodulating, near-isogenic lines altered in the nts locus, which controls nodule formation, could be distinguished from each other and from the parent cultivar by amplification of template pre-digested with 2-3 restriction enzymes. A total of 42 DNA polymorphisms were detected using only 19 octamer primers. In the absence of digestion, 25 primers failed to differentiate these soybean genotypes. Several polymorphic products co-segregated tightly with the nts locus in F2 families from crosses between the allelic mutants nts382 and nts1007 and the ancestral G. soja Sieb. & Succ. PI468.397. Our results suggest that EMS is capable of inducing extensive DNA alterations, probably around discrete mutational hot-spots. EMS-induced DNA polymorphisms may constitute sequence tagged markers diagnostic of specific genomic regions. PMID- 8232213 TI - Nopaline causes a conformational change in the NocR regulatory protein-nocR promoter complex of Agrobacterium tumefaciens Ti plasmid pTiT37. AB - The nocR gene of Agrobacterium tumefaciens Ti plasmid pTiT37 is the regulatory gene of the nopaline catabolism (noc) operon of pTiT37. We have cloned and sequenced nocR, which encodes a DNA-binding protein. The deduced amino acid sequence is similar to those of members of the LysR family of prokaryotic activator proteins. Gel retardation experiments demonstrated that the NocR protein binds to the nocR promoter in both the presence and absence of nopaline. The increased mobility of the complex and alterations in the DNase I footprints revealed a nopaline-induced conformational change in the NocR-DNA complex. Sequence analysis of the NocR binding site indicated the presence immediately downstream of the -10 sequence of the nocR promoter of a 12 bp putative operator overlapping a consensus gyrase recognition sequence and an 18 bp long alternating purine-pyrimidine sequence. These results suggest that nopaline-induced alterations in the NocR protein-nocR promoter complex might control gene expression in the noc operon. PMID- 8232214 TI - Transient and stable gene expression in the fungal maize pathogen Cochliobolus heterostrophus after transformation with the beta-glucuronidase (GUS) gene. AB - The bacterial GUS (beta-glucuronidase) gene has been used as a reporter gene in plants and bacteria and was recently expressed in filamentous fungi. Here, we report the application of GUS for the establishment of transient and stable gene expression systems in the phytopathogenic fungus Cochliobolus heterostrophus. The utility of the transient expression system is demonstrated in applications involving promoter analysis and in tests of various parameters of a transformation system, for comparing the rates of stable and transient transformation events using GUS as sole screening marker and for comparing different transformation systems using either GUS or a dominant selection marker. For these purposes two plasmids were constructed harbouring the GUS gene and the hph gene of Escherichia coli which confers resistance to the antibiotic hygromycin B (HygB), ligated either to the P1 or GPD1 (glyceraldehyde 3 phosphate dehydrogenase) promoter of C. heterostrophus. In transient expression studies the first appearance of GUS activity was observed within 2 h after transformation and maximal values were obtained after 7 or 10 h, depending on the promoter fused to the GUS gene. At peak activity, the GPD1 promoter was revealed to be five fold stronger than the P1 promoter. The same difference in promoter strength was observed when the vectors were stably integrated in the fungal genome. Using the GUS gene as a colour selection marker in plate assays, it was possible to detect transformants and monitor the process of transient gene expression visually. Blue transformants obtained by screening for the GUS phenotype were mitotically unstable. Transformants obtained by selecting for HygB resistance were mitotically stable and expressed the beta-glucuronidase gene constitutively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232215 TI - Genetic analysis of yeast strains lacking negative feedback control: a one-step method for positive selection and cloning of carbamoylphosphate synthetase aspartate transcarbamoylase mutants unable to respond to UTP. AB - We have undertaken an in vivo genetic approach to the analysis of negative feedback control by uridine triphosphate (UTP) of the yeast carbamoylphosphate synthetase-aspartate transcarbamoylase multifunctional protein (CPSase-ATCase). Using an analog of uracil, 5-fluorouracil, we have constructed a screening system leading, in one step, to selection and cloning of a functional aspartate transcarbamoylase that is defective in negative feedback control by UTP. Due to the nature of the screen, spontaneous or UV-induced mutants could be recovered. Well-characterized cloned mutants have been sequenced and reveal one or two modifications in single codons leading to single amino acid replacements. These amino acid changes occurred either in the CPSase or ATCase domains, abolishing their sensitivity to regulation but not their catalytic activities. Hence the regulatory and catalytic sites are distinct. With the same screening system, it may also be possible to enlarge the scope of the molecular study of the feedback processes to include equivalent proteins in fungi as well as higher eukaryotes. PMID- 8232216 TI - Oligomerization of Escherichia coli haemolysin (HlyA) is involved in pore formation. AB - Coexpression of pairs of nonhaemolytic HlyA mutants in the recombination deficient (recA) strain Escherichia coli HB101 resulted in a partial reconstitution of haemolytic activity, indicating that the mutation in one HlyA molecule can be complemented by the corresponding wild-type sequence in the other mutant HlyA molecule and vice versa. This suggests that two or more HlyA molecules aggregate prior to pore formation. Partial reconstitution of the haemolytic activity was obtained by the combined expression of a nonhaemolytic HlyA derivative containing a deletion of five repeat units in the repeat domain and several nonhaemolytic HlyA mutants affected in the pore-forming hydrophobic region. The simultaneous expression of two inactive mutant HlyA proteins affected in the region at which HlyA is covalently modified by HlyC and the repeat domain, respectively, resulted in a haemolytic phenotype on blood agar plates comparable to that of wild-type haemolysin. However, complementation was not possible between pairs of HlyA molecules containing site-directed mutations in the hydrophobic region and the modification region, respectively. In addition, no complementation was observed between HlyA mutants with specific mutations at different sites of the same functional domain, i.e. within the hydrophobic region, the modification region or the repeat domain. The aggregation of the HlyA molecules appears to take place after secretion, since no extracellular haemolytic activity was detected when a truncated but active HlyA lacking the C terminal secretion sequence was expressed together with a nonhaemolytic but transport-competent HlyA mutant containing a deletion in the repeat domain. PMID- 8232217 TI - Replication of the promiscuous plasmid pLS1: a region encompassing the minus origin of replication is associated with stable plasmid inheritance. AB - Deletion of a region of the promiscuous plasmid pLS1 encompassing the initiation signals for the synthesis of the plasmid lagging strand led to plasmid instability in Streptococcus pneumoniae and Bacillus subtilis. This defect could not be alleviated by increasing the number of copies (measured as double-stranded plasmid DNA) to levels similar to those of the wild-type plasmid pLS1. Our results indicate that in the vicinity of, or associated with the single-stranded origin region of pLS1 there is a plasmid component involved in its stable inheritance. Homology was found between the DNA gyrase binding site within the par region of plasmid pSC101 and the pLS1 specific recombination site RSB. PMID- 8232218 TI - Structure and properties of omega-agatoxin IVB, a new antagonist of P-type calcium channels. AB - A new peptide antagonist of voltage-activated calcium channels was purified from venom of the funnel web spider, Agelenopsis aperta. This 48-amino acid peptide, omega-agatoxin (omega-Aga)-IVB, was found to be a potent (Kd, approximately 3 nM) blocker of P-type calcium channels in rat cerebellar Purkinje neurons but had no activity against T-type, L-type, or N-type calcium channels in a variety of neurons. The calcium channel-blocking properties of omega-Aga-IVB were similar to those of another toxin, omega-Aga-IVA, which has 71% amino acid identity with omega-Aga-IVB. The 10-fold greater abundance of omega-Aga-IVB in venom allowed structural studies using NMR spectroscopy. The three-dimensional structure derived from NMR data resulted in a proposed disulfide bond configuration for the peptide. Although omega-Aga-IVB has fewer basic and more acidic residues than does omega-Aga-IVA, the two toxins show conservation of positively charged residues in a mid-peptide region that is predicted to form one face of the omega Aga-IVB molecule. This region may be crucial for high affinity binding to the P type calcium channel. In contrast, the amino termini of the two toxins have different charges and seem unlikely to be involved in binding to the channel. PMID- 8232219 TI - Anti-human immunodeficiency virus type 1 therapy and peripheral neuropathy: prevention of 2',3'-dideoxycytidine toxicity in PC12 cells, a neuronal model, by uridine and pyruvate. AB - A strategy for preventing or delaying the peripheral neuropathy induced by 2',3' dideoxycytidine (ddC) therapy in patients with acquired immunodeficiency syndrome was suggested by findings, in two laboratories, that cultured avian and mammalian cells devoid of mitochondrial DNA continue to replicate at virtually normal rates, provided that the medium is supplemented with uridine and pyruvate. Inasmuch as it is likely that a depletion of mitochondrial DNA also takes place in neuronal cells exposed to ddC, we used PC12 cells, the neuronal model we have reported on previously, in an attempt to rescue these cells from the deleterious effects of ddC. We first show, using undifferentiated PC12 cells, that DNA replication is impaired in mitochondria isolated from cells grown in the presence of ddC. Then, using growth rate as a criterion of the well-being of the cells, we show that the addition of uridine and pyruvate to uninduced cells growing in the presence of ddC results in an average rescue efficiency of 51%, based on the uridine/pyruvate-treated control. This value increases considerably at substantially higher concentrations of uridine alone. Rescue efficiencies of differentiated cells, which do not proliferate, were assessed using neurite outgrowth and neurite survival as criteria. Here the rescue efficiency is 56%, based on the uridine/pyruvate-treated control. In addition, uridine and pyruvate prolong the viability of ddC-treated cells and maintain their healthy appearance; without these compounds, the ddC-treated cells have an abnormal morphology and die off quite rapidly. PMID- 8232220 TI - Cytokines down-regulate expression of major cytochrome P-450 enzymes in adult human hepatocytes in primary culture. AB - Cytokines are thought to cause the depression of cytochrome P-450 (CYP) associated drug metabolism in humans during inflammation and infection. We have examined the role of five cytokines, i.e., interleukin-1 beta, interleukin-4, interleukin-6, tumor necrosis factor-alpha, and interferon-gamma, on the expression of CYP1A2, CYP2C, CYP2E1, CYP3A, and epoxide hydrolase in primary human hepatocyte cultures. Steady state P-450 and epoxide hydrolase mRNA levels, as well as ethoxyresorufin-O-deethylase and nifedipine oxidation activities, which are mainly supported by CYP1A1/1A2 and CYP3A, respectively, were measured. Interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha were found to be the most potent depressors of P-450 enzymes. After 3 days of treatment, both mRNA levels and enzyme activities were depressed, typically by at least 40%, whatever the cytokine and the enzyme considered. Interferon-gamma also suppressed CYP1A2 and CYP2E1 mRNA levels and ethoxyresorufin-O-deethylase activity but had no effect on CYP3A and epoxide hydrolase mRNAs. In addition, interleukin-4 had the opposite effect, compared with other cytokines, on CYP2E1 mRNA, which was increased up to 5-fold; ethoxyresorufin-O-deethylase and nifedipine oxidation activities were not significantly affected. These results provide the first demonstration that various cytokines act directly on human hepatocytes to affect expression of major P-450 genes and that a wide range of responses can be observed among the enzymes for a given cytokine, suggesting that different regulatory mechanisms may be involved. PMID- 8232221 TI - Drosophila nervous system muscarinic acetylcholine receptor: transient functional expression and localization by immunocytochemistry. AB - The pharmacological properties of a cloned Drosophila muscarinic acetylcholine receptor (mAChR) were investigated using two independent transient expression systems. The binding characteristics of the expressed receptor were determined using transfected COS-7 cells, whereas the mAChR functional properties were analyzed using nuclearly injected Xenopus oocytes. Competition displacement studies with transfected COS-7 cell membranes showed that N-[3H]methylscopolamine binding was displaced most effectively by atropine, followed by 4-diphenylacetoxy N-methylpiperidine methiodide, pirenzepine, and AFDX-116. This same order of effectiveness (4-diphenylacetoxy-N-methylpiperidine methiodide > pirenzepine > AFDX-116) was observed in oocytes expressing Dm1 when carbamylcholine-induced currents were inhibited by the same antagonists. Thus, the expressed Drosophila mAChR (Dm1) exhibits a pharmacology that broadly resembles that of the vertebrate M1 and M3 mAChR subtypes. To determine the anatomical localization of the Drosophila mAChR, polyclonal antiserum was raised against a peptide corresponding to the predicted carboxyl-terminal domain of the receptor. Immunocytochemistry on fly sections demonstrated that the mAChR gene product was found in the nervous system and was not seen in skeletal muscle. The most intense staining was localized to the glomeruli of the antennal lobes, an area of the insect brain where first-order synaptic processing of olfactory information occurs. PMID- 8232222 TI - 5-hydroxytryptamine1C receptor density and mRNA levels in choroid plexus epithelial cells after treatment with mianserin and (-)-1-(4-bromo-2,5 dimethoxyphenyl)-2-aminopropane. AB - 5-Hydroxytryptamine (5HT)1C and 5HT2 receptors display paradoxical down regulation when exposed to receptor antagonists in vivo, a property that is unique to these two subtypes of serotonin (5HT) receptors. Because of the absence of cell culture model systems, the mechanisms involved in this paradoxical down regulation have been difficult to explore. The present study focuses on the regulation of 5HT1C receptors in primary cultures of rat choroid plexus epithelial cells. Exposure of the epithelial cell cultures to 100 nM mianserin, a receptor antagonist, or (-)-1-(4-bromo-2,5-dimethoxyphenyl)-2-aminopropane, an agonist, for 72 hr caused a loss of 5HT1C receptor binding sites, as determined by [3H]mesulergine binding to crude membrane preparations. No significant changes in Kd values were observed. Neither the agonist nor antagonist caused a significant change in binding sites after 24 hr. A solution hybridization assay was used to determine whether the down-regulation by mianserin or (-)-1-(4-bromo 2,5-dimethoxyphenyl)-2-aminopropane was accompanied by a decrease in the steady state level of 5HT1C receptor mRNA. These studies showed that neither treatment caused an alteration in the levels of 5HT1C receptor mRNA. Thus, it is possible to reproduce the in vivo regulatory effects of drugs on 5HT1C receptors in choroid plexus epithelial cells in culture, including the atypical down regulation by receptor antagonists. Using this cell culture model system, indirect transynaptic effects and decreases in receptor mRNA levels have been ruled out as mechanisms accounting for the down-regulation. PMID- 8232223 TI - Modification of G protein-coupled functions by low-pH pretreatment of membranes from NG108-15 cells: increase in opioid agonist efficacy by decreased inactivation of G proteins. AB - Low-pH pretreatment increases opioid agonist efficacy in inhibiting adenylyl cyclase in brain membranes. The mechanism of this effect was examined in membranes from cultured NG108-15 cells. Pretreatment of NG108-15 membranes at pH 4.5 before assay at pH 7.4 produced the following modifications in G protein mediated signal transduction: 1) decreased activation of adenylyl cyclase by Gs, 2) increased maximal inhibition of opioid agonist binding by sodium and by guanine nucleotides in the presence of sodium, and 3) increased maximal inhibition of adenylyl cyclase by agonists for G(i)-coupled receptors. These results are similar to those previously observed in rat brain membranes. The mechanism by which low-pH pretreatment increased receptor-mediated inhibition of adenylyl cyclase was investigated further by examining low-Km GTPase activity in low-pH-pretreated NG108-15 cell membranes. Low-pH pretreatment decreased basal and agonist-stimulated low-Km GTPase activity maximally in the absence of sodium and minimally in the presence of 120 mM NaCl. This change was due to a decrease in the Vmax of the enzyme, with no change in the Km for GTP, indicating that GTP hydrolysis was decreased without any decrease in the affinity of the G protein for GTP. Scatchard analysis revealed no decrease in the Bmax for high affinity opioid agonist binding, and Western blot analysis with a G(i)-specific antibody revealed no loss of G(i) protein, in low-pH-pretreated membranes. Moreover, concentration-effect curves for GTP in supporting opioid inhibition of adenylyl cyclase showed that low-pH pretreatment increased inhibition by the agonist only at GTP concentrations equal to or greater than the Km for GTP hydrolysis by the low-Km GTPase. Taken together, these results indicate that the efficacy of receptor-mediated inhibition of adenylyl cyclase can be increased by decreasing the maximal inactivation rate of G(i) subsequent to its activation by the receptor. PMID- 8232224 TI - Biochemical and immunochemical comparison of saxiphilin and transferrin, two structurally related plasma proteins from Rana catesbeiana. AB - Saxiphilin is a approximately 90-kDa protein in bullfrog plasma that binds the neurotoxin saxitoxin (STX) with high affinity (Kd, approximately 0.2 nM). The relationship between saxiphilin and transferrin was examined because partial sequencing of saxiphilin previously revealed an unexpected homology to members of the transferrin family of Fe(3+)-binding proteins. Transferrin was purified from bullfrog plasma and shown to be distinct from saxiphilin on the basis of its size (approximately 78 kDa), chromatographic behavior, visible absorption spectrum, and ligand-binding properties. High affinity binding of [3H]STX was found to be a distinctive property of saxiphilin that was not exhibited by transferrins from various species of animals. Conversely, under conditions appropriate for transferrins, purified saxiphilin did not bind 55Fe3+, implying that it is not involved in iron metabolism. Polyclonal antibodies raised against native saxiphilin precipitated [3H]STX-binding activity from whole bullfrog plasma. On immunoblots such antibodies recognized the denatured saxiphilin protein but only weakly labeled bullfrog transferrin. In an enzyme-linked immunosorbent assay using native proteins, antisaxiphilin antibodies weakly cross-reacted with transferrin from bullfrog and a number of other species. Likewise, antibodies against human transferrin cross-reacted with saxiphilin in a similar immunosorbent assay. These results lead to the conclusion that saxiphilin is not bullfrog transferrin but is structurally related to the transferrin family. As a novel member of the transferrin superfamily, saxiphilin may help to uncover new functions mediated by this class of proteins. PMID- 8232225 TI - Beta-[3H]funaltrexamine-labeled mu-opioid receptors: species variations in molecular mass and glycosylation by complex-type, N-linked oligosaccharides. AB - We previously showed that under defined conditions beta-[3H]funaltrexamine (beta [3H]FNA) covalently labeled mu-opioid receptors with high specificity in bovine striatal membranes. beta-[3H]FNA-labeled mu-opioid receptors migrated as a broad band with a molecular mass range of 68-97 kDa. It is controversial whether beta FNA binds irreversibly to mu-opioid receptors in other species. In this study, we demonstrated that beta-[3H]FNA also labeled mu-opioid receptors with high specificity in brain membranes of the guinea pig, rat, and mouse. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography revealed that in each species beta-[3H]FNA specifically bound to a protein in which labeling was greatly reduced by naloxone. These labeled receptors had broad molecular mass ranges, and the molecular masses were different among these species, in the order of cow > guinea pig > rat > mouse. Membranes were subjected to solubilization with 2% Triton X-100 and wheat germ lectin (WGL) affinity chromatography. N Acetylglucosamine eluted a peak of radioactivity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and fluorography showed that in all four species the mu receptor was the only protein labeled with beta-[3H]FNA in the WGL eluate. The molecular masses of labeled mu-opioid receptors were 70-88 kDa (median, 77 kDa) for the cow, 66-80 kDa (median, 72 kDa) for the guinea pig, 60 75 kDa (median, 67 kDa) for the rat, and 60-72 kDa (median, 66 kDa) for the mouse. In addition, we investigated the nature of the carbohydrate moieties linked to the receptor protein and whether the species variation in the molecular mass was due to variable degrees of glycosylation. The bovine WGL eluate was treated with various glycosidases. Neuraminidase treatment decreased the receptor molecular mass by 6-7 kDa, whereas alpha-mannosidase had no effect. Removal of N linked carbohydrates at asparagine residues by peptide-N4-[N-acetyl-beta glucosaminyl]asparagine amidase (N-Glycanase) resulted in a much sharper specifically labelled protein band of 43 kDa. These results indicate that mu opioid receptors are heavily glycosylated and the major carbohydrate moieties are of the complex type, N-linked to asparagine. After the WGL eluates for the four species were treated with N-Glycanase, the labeled receptors became much sharper bands with very similar molecular masses, i.e., 43 kDa for the cow and guinea pig, 39 kDa for the rat, and and 40 kDa for the mouse.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8232226 TI - Spongistatin 1, a highly cytotoxic, sponge-derived, marine natural product that inhibits mitosis, microtubule assembly, and the binding of vinblastine to tubulin. AB - A highly cytotoxic macrocyclic lactone polyether has been isolated from a Spongia species and named spongistatin 1. With L1210 murine leukemia cells an IC50 value for cell proliferation of 20 pM was obtained, and an increase in the mitotic index concordant with the decrease in cell number was observed. Kangaroo rat kidney PtK1 cells were examined by indirect immunofluorescence with a spongistatin 1 concentration that caused 50% reduction in cellular protein (0.3 nM) and with a 10-fold higher concentration. These cells displayed mitotic and nuclear aberrations at both concentrations, and intracellular microtubules were reduced in number at the lower concentration and disappeared at the higher. Similar changes in PtK1 cells were observed after treatment with equivalent toxic concentrations of the antimitotic agents colchicine, vinblastine, halichondrin B, and dolastatin 10. Spongistatin 1 inhibited the glutamate-induced polymerization of purified tubulin (IC50 value of 3.6 microM versus 2.1 microM for dolastatin 10 and vinblastine and 5.2 microM for halichondrin B). Spongistatin 1 had no effect on the binding of colchicine to tubulin, but it was a potent inhibitor of the binding of vinblastine and GTP to tubulin. In initial experiments with 5 microM tubulin and 5 microM vinblastine, spongistatin 1 and dolastatin 10 both had IC50 values of 2 microM, whereas halichondrin B had an IC50 value of 5 microM. Spongistatin 1 thus represents a new member of the group of complex natural products that inhibit mitosis by binding in the Vinca alkaloid domain of tubulin. PMID- 8232227 TI - Dual topoisomerase I and II inhibition by intoplicine (RP-60475), a new antitumor agent in early clinical trials. AB - The mechanisms of action of intoplicine (RP-60475), a 7H-benzo[e]pyrido[4,3 b]indole derivative that is presently in early clinical trials, have been investigated. Intoplicine induced both topoisomerase I- and II-mediated DNA strand breaks, using purified topoisomerases. The topoisomerase cleavage site patterns induced by intoplicine were unique, relative to those of camptothecin, 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA), and other known topoisomerase inhibitors. Both topoisomerase I- and II-induced DNA breaks decreased at drug concentrations higher than 1 microM, which is consistent with the DNA-intercalating activity of intoplicine. DNA damage was investigated in KB cells in culture by using alkaline elution. Intoplicine induced single-strand breaks (SSB) in a bell-shaped manner with respect to drug concentration (maximum frequency at 1 microM approximately 220 rad-equivalents). SSB formation was fast, whereas reversal after drug removal was slow. Similar bell-shaped curves were obtained for DNA double-strand breaks (DSB) and DNA-protein cross-links. SSB and DNA-protein cross-link frequencies were approximately equal, and no protein-free breaks were detectable, indicating the protein concealment of the breaks, as expected for topoisomerase inhibition. Comparison of SSB and DSB frequencies indicated that intoplicine produced a significant amount of SSB not related to DSB, which is consistent with concomitant inhibition of both DNA topoisomerases I and II in cells. Data derived from resistant cell lines indicated that multidrug resistant cells were cross-resistant to intoplicine but that m-AMSA- and camptothecin-resistant cells were sensitive to intoplicine. Hence, intoplicine might circumvent topoisomerase I-mediated and topoisomerase II-mediated resistance by poisoning both enzymes simultaneously. PMID- 8232228 TI - The phenobarbital-induced transcriptional activation of cytochrome P-450 genes is blocked by the glucocorticoid-progesterone antagonist RU486. AB - Several of the hepatic microsomal cytochromes P450 can be induced by various drugs and xenobiotics, among them the barbiturate phenobarbital. Rat hepatoma cells (Fao and its derivatives) respond to phenobarbital or dexamethasone treatment with an increased accumulation of CYP2C6 mRNA and thus provide a culture system to investigate the mechanisms involved. Examination of the kinetics of CYP2C6 mRNA induction revealed that the response to dexamethasone is rapid, whereas induction by phenobarbital occurs only slowly after an 8-10-hr lag. Run-on transcription measurements demonstrated that phenobarbital treatment led to a 3-4-fold increase in CYP2C6 gene transcription. Surprisingly, induction by phenobarbital of both accumulation of CYP2C6 mRNA and transcription of the gene was blocked by the antiprogestin-antiglucocorticoid RU486, suggesting the involvement of a steroid receptor in the induction process. Transfection of promoter constructs containing a reporter gene whose expression is driven by a 1.4-kilobase 5' flanking segment of the CYP2B1 or CYP2B2 genes, which are highly inducible by phenobarbital in rat liver, led to > 3-fold increases in reporter gene activity in the presence of the drug. Again, phenobarbital induction was prevented by RU486. The RU486 inhibition of the phenobarbital induction of both the endogenous CPY2C6 gene and the transfected CYP2B1 and CYP2B2 promoter constructs leads us to propose a model whereby the drug acts indirectly to cause the accumulation of an endogenous steroid, and this molecule, acting via its receptor, would be the direct inducer of cytochromes P450. Whether or not this model proves to be correct, the results presented here provide the first evidence of the involvement of a steroid receptor in phenobarbital induction. PMID- 8232229 TI - Coupling of expressed alpha 1B- and alpha 1D-adrenergic receptor to multiple signaling pathways is both G protein and cell type specific. AB - alpha 1-Adrenergic receptors (ARs) are members of the G protein-coupled receptor superfamily. alpha 1-AR subtypes mediate the effects of the sympathetic nervous system, especially those involved in cardiac homeostasis. To investigate signal transduction by a novel subtype (alpha 1D), which we recently cloned, and to compare it with that by the previously characterized alpha 1B-AR, we assessed the ability of each subtype to activate polyphosphoinositide (PI) metabolism, cAMP accumulation, and arachidonic acid release in Chinese hamster ovary (CHO) and COS 1 cells expressing these subtypes after stable or transient transfection, respectively. In COS-1 and CHO cells, both the alpha 1D- and alpha 1B-AR were found to couple to PI hydrolysis through a pertussis toxin-insensitive G protein. Both alpha 1-AR subtypes also increased intracellular cAMP by an indirect mechanism, although this effect was observed only in COS-1 cells and not in CHO cells. Interestingly, alpha 1-AR-stimulated arachidonic acid release was also demonstrated for both subtypes in COS-1 cells. This release was mediated through phospholipase A2 activation and involved a pertussis toxin-sensitive G protein. alpha 1-AR-stimulated arachidonic acid release was dependent upon extracellular calcium and was inhibited by 1 microM nifedipine. Inhibitors of protein kinase C, phospholipase C, and diacylglycerol lipase did not alter alpha 1-AR-stimulated release of arachidonic acid. These findings indicate that in COS-1 cells alpha 1 AR-stimulated arachidonic acid release is most likely coupled to dihydropyridine sensitive L-type calcium channels via a pertussis toxin-sensitive G protein. The influx of extracellular calcium then stimulates phospholipase A2 to release arachidonic acid. alpha 1-AR-stimulated arachidonic acid release could also be demonstrated in CHO cells and was pertussis toxin sensitive but nifedipine insensitive. These cells were also unresponsive to Bay K8644, indicating a lack of voltage-sensitive calcium channels in CHO cells. Nevertheless, alpha 1-AR activation increased intracellular Ca2+ levels, as assessed by fura-2 fluorescence studies. Neomycin blocked both alpha 1-AR-stimulated PI hydrolysis and increases in intracellular Ca2+ levels but did not inhibit the increase in arachidonic acid release. Taken together, these data indicate that in CHO cells alpha 1-ARs can couple directly to phospholipase A2 activation via a pertussis toxin-sensitive pathway. Thus, in these model systems we demonstrate for the first time that a single alpha 1-AR subtype can activate multiple distinct signal transduction pathways, in which receptor-effector coupling is modulated by distinct G proteins. PMID- 8232230 TI - Antisera against peptides derived from a purified mu-opioid binding protein recognize the protein as well as mu-opioid receptors in brain regions and a cell line. AB - Two peptides, which have no significant homology with known protein structures, were obtained by microsequencing of a mu-opioid binding protein purified to homogeneity from bovine striatal membranes. Polyclonal antibodies generated against portions of these peptides immunoprecipitated up to 65% of radiolabeled purified opioid binding protein. Sequential immunoprecipitations, using antibodies directed against portions of the two different peptides, confirmed that the peptides are derived from the same protein. Immunoblots of the protein with antipeptide antibodies revealed a protein band corresponding to the molecular weight of denatured reduced mu-opioid binding protein. The immunoresponse was blocked by the appropriate peptide and was not observed with irrelevant antisera. The antipeptide antibodies were used for immunoblots of sodium dodecyl sulfate extracts of tissues from bovine brain regions and of the mu receptor-containing cell line SK-N-SH. Affinity-purified antipeptide antibody detected an immunoreactive protein of molecular weight 65,000 in brain regions containing high levels of mu-opioid receptors (striatum, thalamus, hippocampus, and frontal cortex) and in the cell line SK-N-SH. Pons, which contains low levels of receptors, produced a a barely detectable signal, whereas white matter, HeLa cells, and C6 glioma cells, devoid of opioid binding activity, produced no detectable signal. The correlation between immunoreactivity and the presence of mu-opioid binding in brain regions and cell lines and the correspondence of the molecular weight of the immunoreactive protein to that of mu-opioid receptors provide strong evidence that the peptide antisera recognize mu receptors. PMID- 8232231 TI - Selective coupling of alpha 2-adrenergic receptor subtypes to cyclic AMP dependent reporter gene expression in transiently transfected JEG-3 cells. AB - A cAMP-dependent reporter gene has been used in transiently transfected human choriocarcinoma (JEG-3) cells to examine the second messenger coupling of the human alpha 2-adrenergic receptor subtypes. The reporter gene consists of a cAMP response element linked to the gene for chloramphenicol acetyltransferase (CAT). Plasmids encoding the alpha 2-C10 (alpha 2A), alpha 2-C2 (alpha 2B), or alpha 2 C4 (alpha 2C) receptor subtypes were co-transfected with a plasmid containing the reporter gene, and the ability of alpha 2 receptor agonists to influence forskolin-stimulated CAT expression was examined. For alpha 2-C10, agonists had a biphasic effect on forskolin-stimulated CAT expression. Thus, low (nanomolar) concentrations of agonist inhibited CAT expression by approximately 60%, whereas high (micromolar) concentrations reversed this inhibition and could even potentiate CAT expression by as much as 140%. A significantly different pattern of coupling was observed for the other alpha 2 receptor subtypes. For alpha 2-C4, agonists only inhibited forskolin-stimulated CAT expression, whereas for alpha 2 C2 only potentiation of expression was seen. Each of these responses was specifically blocked by alpha 2- but not alpha 1- or beta-adrenergic receptor antagonists. For alpha 2-C4, the inhibition of forskolin-stimulated CAT expression was prevented by pretreatment of the cells with pertussis toxin. This was also true for the inhibition obtained with alpha 2-C10. The potentiation of CAT expression, however, was not prevented by pertussis toxin pretreatment in cells transfected with either alpha 2-C2 or alpha 2-C10. In this transient expression system, each alpha 2-adrenergic receptor subtype had access to the same complement of G proteins, adenylyl cyclase, and other second messengers. It would appear, therefore, that the potential for the activation of unique intracellular responses exists even among closely related receptor subtypes. PMID- 8232232 TI - Inositol-1,3,4,5-tetrakisphosphate induces calcium mobilization via the inositol 1,4,5-trisphosphate receptor in SH-SY5Y neuroblastoma cells. AB - myo-Inositol-1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P4]-induced Ca2+ mobilization was examined in saponin-permeabilized SH-SY5Y cells using myo-inositol hexakisphosphate-supplemented buffer to prevent Ins(1,3,4,5)P4-3-phosphatase catalyzed back-conversion of exogenous Ins(1,3,4,5)P4 to myo-inositol-1,4,5 trisphosphate [Ins(1,4,5)P3]. The Ins(1,3,4,5)P4 concentration-response curve for Ca2+ release in SH-SY5Y cells exhibited an EC50 of 2.5 microM, compared with 52 nM for Ins(1,4,5)P3, with the maximally effective concentration of Ins(1,3,4,5)P4 (100 microM) mobilizing the entire Ins(1,4,5)P3-sensitive pool. Both Ins(1,3,4,5)P4- and Ins(1,4,5)P3-induced Ca2+ mobilizations were heparin sensitive. Further, L-chiro-inositol-2,3,5-trisphosphorothioate, a recently identified low intrinsic activity Ins(1,4,5)P3 receptor partial agonist, shifted both the Ins(1,4,5)P3 and Ins(1,3,4,5)P4 concentration-response curves significantly rightward, with similar potencies. However, binding studies demonstrate that L-chiro-inositol-2,3,5-trisphosphorothioate interacts very poorly (IC50 > 30 microM) with specific Ins(1,3,4,5)P4 binding sites that have been previously characterized in pig cerebellum. Carbachol-pretreated SH-SY5Y cells (1 mM, > or 6 hr) exhibit a decrease in Ins(1,4,5)P3 receptor number, accompanied by both a rightward shift and a reduced maximal Ca2+ release in their Ins(1,4,5)P3 concentration-response curve. Here both Ins(1,4,5)P3 and Ins(1,3,4,5)P4 concentration-response curves were found to exhibit identically reduced maximal Ca2+ release responses and about 4-fold rightward shifts in EC50 values. Together, these observations provide compelling evidence for our hypothesis that Ins(1,3,4,5)P4 exhibits weak but full agonist status at Ins(1,4,5)P3 receptor-operated Ca2+ channels in SH-SY5Y cells. PMID- 8232233 TI - BW373U86: a nonpeptidic delta-opioid agonist with novel receptor-G protein mediated actions in rat brain membranes and neuroblastoma cells. AB - BW373U86 is a potent and highly selective nonpeptidic agonist for delta-opioid receptors. To determine its ability to couple with G protein-linked second messenger systems, this study examined the effects of BW373U86 on the inhibition of adenylyl cyclase and the stimulation of low-Km GTPase activity. In rat striatal membranes, BW373U86 inhibited basal adenylyl cyclase activity in a GTP dependent manner, with maximal inhibition levels similar to those of the prototypic delta agonist [D-Ser2,Thr6]Leu-enkephalin (DSLET). However, BW373U86 was approximately 100 times more potent than DSLET in inhibiting adenylyl cyclase. Analysis of the inhibitory activity across 10 brain regions revealed that both low and high concentrations of BW373U86 inhibited adenylyl cyclase activity in a manner similar to that of DSLET. Inhibition of adenylyl cyclase by BW373U86 was delta receptor selective, because the delta receptor-selective antagonist naltrindole was significantly more potent than naloxone and the mu receptor-selective antagonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 was ineffective in blocking BW373U86 inhibition. BW373U86 also inhibited adenylyl cyclase activity in membranes prepared from NG108-15 cells, with an IC50 value 5 times lower than that of DSLET. This increased potency was not observed in concentration-effect curves for agonist-stimulated low-Km GTPase in NG108-15 membranes. BW373U86 is a competitive inhibitor of [3H]diprenorphine at delta receptors of NG108-15 cell membranes. However, unlike DSLET, BW373U86 displacement of [3H]diprenorphine binding to NG108-15 cell membranes was not affected by sodium and guanine nucleotides. This lack of GTP effect on binding apparently produced slow dissociation rates for this agonist, because naltrindole was less potent in blocking BW373U86 inhibition of adenylyl cyclase when membranes were preincubated with this agonist. These results demonstrate the novel finding that the binding of a full agonist to a G protein-coupled receptor is not regulated by GTP, and they also show how the lack of regulation in receptor binding affects agonist potency. PMID- 8232234 TI - A selective adenosine antagonist (8-cyclopentyl-1,3-dipropylxanthine) eliminates both neuromuscular depression and the action of exogenous adenosine by an effect on A1 receptors. AB - The effect of 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), a selective A1 adenosine receptor antagonist, was studied at frog motor nerve endings in the hope of determining whether the inhibitory effects of exogenous or endogenous adenosine on neurotransmitter release are mediated by an A1 receptor or the postulated prejunctional "A3 receptor." These putative A3 receptors have been reported to have a lower affinity for DPCPX (>> 1 nM) than A1 receptors (50-190 pM) and have been linked to changes in Ca2+ translocation. The affinity of DPCPX as an antagonist of exogenous adenosine at frog motor nerve endings was calculated by using the Schild equation and found to range from 25 to 200 pM (n = 12). These values are consistent with the presence of A1 receptors. The effect of endogenous adenosine as a mediator of prejunctional neuromuscular depression produced by repetitive nerve impulses was fully reversed by 100 pM DPCPX. Neither prejunctional neuromuscular depression produced by endogenous or exogenous adenosine nor the reversal of depression by DPCPX was associated with changes in nerve terminal Ca2+ currents. The results demonstrate that endogenous or exogenous adenosine mediates neuromuscular depression in the frog, via an A1 receptor. PMID- 8232235 TI - U-93631 causes rapid decay of gamma-aminobutyric acid-induced chloride currents in recombinant rat gamma-aminobutyric acid type A receptors. AB - We discovered the ability of U-93631 (4-dimethyl-3-t-butylcarboxyl-4,5- dihydro[1,5-a]imidazoquinoxaline) to accelerate decay of gamma-aminobutyric acid (GABA)-induced currents, and we explored its mechanism in human embryonic kidney cells (HEK-293) stably expressing the alpha 1 beta 2 gamma 2 subtype of GABAA receptors. Inward currents (Cl- efflux) induced by 5 microM GABA at the holding potential of -60 mV (under a symmetrical Cl- gradient) decayed with an exponential time course with a mean time constant (tau) of 222 +/- 25 sec, as examined with the whole-cell configuration of the patch-clamp technique. The monoexponential decay was greatly accelerated in the presence of U-93631 at 5 microM, with the mean tau value being 5.2 +/- 0.5 sec. The tau values were dependent on the concentration of U-93631, with an estimated Kd of approximately 2 microM. Outward currents at the holding potential of +60 mV decayed with a similar tau value in the presence of the drug, suggesting the voltage independence of the drug action. The initial amplitude of the GABA (5 microM) induced Cl- current was not affected by preincubation with U-93631 (5 microM) or GABA (200 nM) alone but was reduced by preincubation with the combination of the two. In the presence of U-93631 at 5 microM, the peak amplitude decreased as a function of GABA concentration, with the half-maximal inhibitory concentration being approximately 100 nm, which is close to the Kd for the high affinity GABA site (85 nM). It appears that the drug interacts with GABA-bound receptors (at least monoliganded) and accelerates receptor desensitization, rather than acting as an open channel blocker. The binding site for U-93631 on GABAA receptors seems not to overlap with GABA, barbiturate, or benzodiazepine sites, because the drug effect persisted in the presence of excess ligands for those sites. With cloned GABAA receptors composed of only alpha 1 beta 2, beta 2 gamma 2, or alpha 1 gamma 2 subunits, U-93631 also accelerated the decay rate. This lack of subtype selectivity raises the possibility that the compound interacts with a region common among the three subunits, probably a novel modulatory site, which can possibly be exploited as a novel therapeutic target. PMID- 8232236 TI - Ethanol inhibits a neuronal ATP-gated ion channel. AB - The cellular mechanisms by which ethanol affects nervous system function are poorly understood. However, evidence has been accumulating that ethanol can affect the function of neurotransmitter-gated ion channels. Extracellular ATP has recently been reported to produce excitatory actions in the peripheral and central nervous systems by activating ligand-gated ion channels. We studied the effect of ethanol on membrane ion current activated by extracellular ATP in isolated bullfrog dorsal root ganglion neurons, by means of the whole-cell patch clamp technique. The amplitude of the ATP-activated current was decreased by ethanol in a concentration-dependent manner over the range of 3-500 mM. The average inhibition of 1 microM ATP-activated current by 100 mM ethanol was 64 +/- 3%, and the concentration of ethanol that produced 50% inhibition was 68 mM. Ethanol inhibition of ATP-activated current was not dependent on membrane potential from -80 to +40 mV, and ethanol did not change the reversal potential of ATP-activated current. Ethanol (100 or 400 mM) shifted the ATP concentration response curve to the right, increasing the EC50 for ATP from 3.0 microM to 6.0 microM or 22.3 microM, respectively, but did not reduce the maximal response to ATP. The results suggest that ethanol inhibits ATP-activated current by increasing the apparent dissociation constant for the ATP receptor. PMID- 8232237 TI - Zinc selectively inhibits flux through benzodiazepine-insensitive gamma aminobutyric acid chloride channels in cortical and cerebellar microsacs. AB - The effects of Zn2+ on the activity of gamma-aminobutyric acid (GABA)A receptor Cl- ionophore complexes found in adult rat cortex and cerebellum were tested by measuring 36Cl- influx into microsacs. In both preparations, the concentration response curves were biphasic, with 25% of the cerebellar and 20% of the cortical Cl- flux being blocked by less than 10 microM Zn2+ and 45% of the cerebellar and 50% of the cortical flux being blocked by concentrations of Zn2+ exceeding 10 microM. Zn2+ (100 microM) did not affect basal Cl- flux but inhibited that stimulated by 100 microM GABA in a noncompetitive manner. The ability of 1 microM flunitrazepam to enhance Cl- flux was unaffected by 100 microM Zn2+. These results demonstrate that, in adult rat cerebellum and cortex, there are three populations of GABAA receptors, two that are sensitive to Zn2+ and insensitive to benzodiazepines (BDZ) and the remainder that are the reverse, i.e., insensitive to Zn2+ but fully sensitive to BDZ enhancement. This result is consistent with the idea that Zn2+ blocks only those GABAA receptor-Cl- ionophore complexes that lack a gamma subunit, which is required for modulation by BDZ. The results obtained in this study also show that the proportion of Zn(2+)-sensitive GABA receptors is substantial, suggesting that they play an important role in the functioning of the adult central nervous system. PMID- 8232238 TI - Role of threonine 342 in helix 7 of the 5-hydroxytryptamine type 1D receptor in ligand binding: an indirect mechanism for receptor selectivity. AB - Recent mutations of the 5-hydroxytryptamine (5-HT)1B and 5-HT1D receptor subtypes suggest that a threonine in the seventh transmembrane helix may be responsible for the selectivity of these receptors. A molecular dynamics simulation of a three-dimensional model of the 5-HT1D receptor interacting with a selective agonist, sumatriptan, shows that, although Thr342 in helix 7 does not have a direct interaction with sumatriptan, it contributes to the selectivity of this receptor through an indirect mechanism. The hydrogen bond between O gamma-H of Thr342 and the backbone C = O of Phe338 stabilizes a bent conformation of the helix that is formed due to the interaction between sumatriptan and Asp339 at one end and Tyr346 at the other end. The indirect mechanism may explain the small change in the affinity for the selective agonist sumatriptan of the receptor in which Thr342 was mutated to asparagine. PMID- 8232239 TI - New activation model for the histamine H2 receptor, explaining the activity of the different classes of histamine H2 receptor agonists. AB - Recently we developed amthamine [2-amino-5-(2-aminoethyl)-4-methylthiazole]. This cyclic analogue of dimaprit proved to be the most potent and selective histamine H2 receptor agonist of a series of substituted 4- or 5-(2-aminoethyl)thiazoles. Quantum chemical studies on histamine (N pi-H tautomer), dimaprit, and amthamine revealed that, based upon geometries of molecular electrostatic potentials, it is likely that these agonists accept a proton from the proton-donating receptor site on their double-bonded (heteroaromatic) nitrogen atoms. In contrast to reported models, this new model is able to accommodate and explain the agonistic activities of all known (including nontautomeric) histamine H2 receptor agonists. Quantitative structure-activity relationship studies with a series of substituted histamine derivatives and heterocyclic analogues support the presented model, in which the monocations in extended conformation interact with the receptor surface; their affinities correlate with the proton association constants of the heteroaromatic nuclei. The negatively charged anchoring site for the ethylammonium side chain of these agonists in this model is a functional group with a pKa value of 4.17. PMID- 8232240 TI - Chemical properties of carbonic anhydrase IV, the membrane-bound enzyme. AB - The carbonic anhydrase (CA) isozyme (IV) in microsomes is thought to have a dominant role in secretory processes. Using microsomes from bovine kidney and lung (which had the same activity), we have measured the Km and kcat for CO2 hydration and compared these numbers with those for CA I (red blood cells and gut), CA II (red blood cells and secretory cells), and CA III (muscle). For kidney CA IV, Km is 10 mM and kcat is 170,000 sec-1 at 0 degree, approaching the rate for CA II but much greater than those for CA I or III. The Ki values for 11 sulfonamides with CA IV were measured and in all cases showed less binding (averaging 17-fold) than to CA II. This is the result of reduction of the association rate constants (k(on)), whereas the dissociation constants of the drug-enzyme complexes (k(off) are similar between CA II and IV. Based on these data, full physiological effects may be expected when inhibition of CA IV is about 99%. Anion inhibition of CA IV is similar to that of CA II and less than that of CA I or CA III. Data are compatible with the proposed role of CA IV in physiological events, i.e., HCO3- formation and secretion at one cell border and H+ separation and excretion at the other. PMID- 8232241 TI - Generation of oxygen free radicals during the metabolism of cyclosporine A: a cause-effect relationship with metabolism inhibition. AB - A better understanding of the mechanism of lipid peroxidation during the metabolism of cyclosporine A (CsA) might help explain the toxicities of this immunosuppressive drug on various organs. Our in vitro work used microsomes prepared from livers of phenobarbital-induced male rats. The incubations (total volume 1ml) also contained a NADPH regenerating system and substrate (i.e., CsA, carbon tetrachloride, or aminopyrine) dissolved in ethanol. Lipid peroxidation was inferred from the presence of malondialdehyde (MDA) which was detected by the thiobarbituric acid assay. The formation of CsA hydroxylated metabolites (AM9 and AM1) was monitored by liquid chromatography. The activity of the microsomal incubation was confirmed by measurements of MDA and formaldehyde production caused by increasing concentrations of CsA, carbon tetrachloride, and aminopyrine. The occurrence of hydroxylated metabolites was not coupled to the production of MDA. Aminopyrine could inhibit MDA production by CsA, but CsA could not reduce the formation of formaldehyde by aminopyrine. Erythromycin, a competitor for the binding site of CsA on cytochrome P450, reduced MDA production by CsA, and CsA inhibited formaldehyde production by erythromycin. Interaction studies with SKF 525A, ketoconazole, superoxide dismutase, catalase, alpha tocopherol, and reduced glutathione confirmed the role of cytochrome P450 and the presence of activated oxygen species as a source of microsomal peroxidation which in return may explain the inhibitory effect of CsA on cytochrome P450 itself. PMID- 8232242 TI - Lactate transport by skeletal muscle sarcolemmal vesicles. AB - Recent studies have indicated that lactate traversal of the sarcolemmal membrane of skeletal muscle could be a carrier mediated process. In the present study, the initial rates of L(+)-lactate flux (Jlact) were measured in highly purified rat hindlimb skeletal muscle sarcolemmal vesicles. Fluxes were determined by the vesicle uptake of L(+)-[U-14C]lactate from the extra-vesicular medium. Jlact was saturable with respect to increasing concentrations of L(+)-lactate. Regression of these data to the Michaelis-Menten equation yielded a Km of 12.5 mM. Jlact was inhibited 81% by 10 mM pyruvate and 83% by 5mM alpha-cyano 4 hydroxycinnamate (p < 0.05), but not by D-lactate, indicating the presence of a stereoselective monocarboxylate transporter in the sarcolemmal membrane. Preincubation of the vesicles with the protein modifier, N-ethylmaleimide (20mM), inhibited Jlact by 86% (p < 0.05). An inhibitor of the inorganic anion exchanger, SITS (1mM), had no effect on Jlact. However, Jlact was markedly sensitive to an inwardly directed proton gradient (p < 0.05), and the flux was more closely related to the concentration of external ionic L(+)-lactate than to the protonated (HLa) form. These studies suggest that skeletal muscle sarcolemmal membranes possess a specific transport system for L-lactate and other monocarboxylates, which has similar properties to the lactate carrier described for several other tissues. PMID- 8232243 TI - Intracellular distribution of hexokinase in rabbit brain. AB - Hexokinase in mammalian brain is particulate and usually considered to be bound to the outer mitochondrial membrane. Investigation of rabbit brain mitochondria prepared either by differential centrifugation and discontinuous density gradient centrifugation has provided evidence that this particulate fraction also contains endoplasmic vesicles and synaptosomes. Solubilization of the bound hexokinase by different combinations of detergents and metabolites has proved the existence of different hexokinase binding sites. Electron microscopic examination of hexokinase location by immuno-gold labelling techniques confirmed that hexokinase is indeed predominantly bound to mitochondria but that a significant proportion is also bound to non-mitochondrial membranes. Attempts to quantify this distribution were unsuccessful since different figures were obtained using anti hexokinase IgG affinity purified on immobilized native or denatured hexokinase. Binding studies of the purified rabbit brain mitochondrial hexokinase to rabbit liver mitochondria and microsomes confirmed that in addition to a binding site on mitochondria there is another binding site on microsomes. The N-terminal sequence of hexokinase has been shown to be important for mitochondria binding and also for microsome binding. These results suggest that the intracellular localization of hexokinase in rabbit brain is not exclusively mitochondrial and that the metabolic role of this enzyme should be reconsidered by including a binding site on the endoplasmic reticulum. PMID- 8232244 TI - Carnitine medium/long chain acyltransferase of microsomes seems to be the previously cloned approximately 54 kDa protein of unknown function. AB - A microsomal protein having N-terminal amino acid sequence SDVLELTDEN, was initially described as a phosphatidyl inositol-specific phospholipase C alpha when its cDNA was cloned (Bennett et al., Nature, 334, 268, 1988). Later, this protein, with an estimated molecular mass of 54 to 60 kDa, was shown to lack the phospholipase activity and instead a protein disulfide oxidoreductase and a thiol protease activities were ascribed to it. Following evidences indicated that the protein in question is the carnitine medium/long chain acyltransferase (CPT) of microsomes that was recently purified as a approximately 54 kDa protein (Murthy and Bieber, Protein Exp. Purif. 3, 75, 1992). First, the N-terminal amino acids of the microsomal CPT showed 100% homology to the sequence described above. Second, during purification of this CPT, the oxidoreductase and the thiol protease activities of the microsomes became separated from the CPT and these other activities were not found in the approximately 900 fold enriched CPT preparations. Third, an antibody to this protein did not immunoprecipitate oxidoreductase of the solubilized microsomal extract but precipitated the CPT. This same protein has been studied by others as the ERp61 (endoplasmic reticulum protein), GRP58 (glucose regulated protein), and HIP-70 (hormone induced protein) but its function was not identified. PMID- 8232245 TI - Hydroxyl radicals mediate injury to endothelium-dependent relaxation in diabetic rat. AB - The purpose of this study was to determine the radical species which mediates the toxic effects of exogenous oxygen-derived free radicals on endothelial function of chronic diabetic rat aorta. Endothelium-dependent relaxation to acetylcholine was impaired in diabetic vessels. Exposure to the exogenous free radical generating system of xanthine plus xanthine oxidase selectively impaired endothelium-dependent relaxation to acetylcholine in control and diabetic aorta with relaxations essentially abolished in diabetic aorta. The loss of relaxation to acetylcholine in diabetic aorta was prevented or attenuated by pretreatment with catalase, dimethylthiourea or desferrioxamine, but not by mannitol or superoxide dismutase. These results suggest that hydroxyl radicals play an important role in the endothelial injury produced by oxygen-derived free radicals in chronic diabetic rat aorta. Furthermore, the site of the injury is likely due to intracellular generation of hydroxyl radicals. PMID- 8232246 TI - Expression of pregnancy-specific beta 1-glycoprotein genes in hematopoietic cells. AB - The presence of PSG in blood cells has been demonstrated by immunohistochemical staining. However, the origin of those proteins is not known. This report examines the expression of the PSG genes in different types of freshly isolated blood cells. RNA isolated from bone marrow and peripheral blood cells of healthy individuals was analyzed for PSG transcripts by reverse transcriptase-polymerase chain reaction using synthetic oligonucleotide primers specific for the PSG genes. The level of expression of the PSG genes in different types of cells exhibited significant individual variation. Trace amounts of PSG transcripts could be detected in polymorphonuclear cells (PMN), monocytes and B lymphocytes while T lymphocytes always contained the highest level of transcript. The expression of PSG genes in the blood cells apparently was not affected by the method of isolation nor by overnight culturing of these cells except in the case when lymphocytes were separated by rosetting with sheep red blood cells. All reported PSG transcripts were detected in blood cells. Both type I and type II transcripts of the PSG genes were detected in blood cells with the exception of type II transcript of PSG5 and PSG11 which were only found in the placenta. Tissue specificity in the expression or alternative splicing of some of the PSG family members was implicated. PMID- 8232247 TI - Purification and characterization of bovine brain calmodulin-dependent protein kinase. II. The significance of autophosphorylation in the regulation of 63 kDa calmodulin-dependent cyclic nucleotide phosphodiesterase isozyme. AB - Bovine brain contains two calmodulin-dependent phosphodiesterase kinases which are separated on Sephacryl S-300 column. One of these kinases has been purified to homogeneity and shown to belong to the calmodulin-dependent protein kinase II family. Phosphorylation of the 63 kDa phosphodiesterase by this purified protein kinase results in the incorporation of 1.0 mol phosphate per mol subunit and an accompanying increase in Ca2+ concentrations required for the phosphodiesterase activation by calmodulin. The protein kinase undergoes autophosphorylation to incorporate 1.0 mol phosphate per mol of subunit of the enzyme and the autophosphorylated enzyme is active, independent of the presence of Ca2+. The autophosphorylation reaction as well as the protein kinase reaction are rendered Ca2+ independent in less than 15 seconds when approximately one mol phosphate per mol protein kinase is incorporated. The result suggests that activation of phosphodiesterase phosphorylation reaction may occur prior to the activation of phosphodiesterase and phosphatase during a cell Ca2+ flux via the protein kinase autophosphorylation mechanism. PMID- 8232249 TI - Effect of pyruvate on rat heart thiol status during ischemia and hypoxia followed by reperfusion. AB - Ischemia or hypoxia followed by reperfusion determine a large release of glutathione from isolated and perfused rat heart. The effects of glucose and/or pyruvate administered during ischemia/reperfusion or hypoxia/reperfusion on the release of cytosolic and mitochondrial glutathione are compared. During ischemia, mitochondrial glutathione is released from the mitochondrion to the cytosol forming a unique pool that leaks out to the interstitial space. Reperfusion causes a large release of total glutathione, particularly from cytosol. Total sulfhydryl groups do not undergo modifications after ischemia, while they appear to decrease upon reperfusion. Pyruvate, which protects the heart by inducing a large recovery of the contractile activity after ischemia, markedly prevents the loss of glutathione. Also total sulfhydryl groups of mitochondria do not undergo significant variation upon ischemia and reperfusion in the presence of pyruvate. During hypoxia, in the absence of glucose, glutathione is mainly lost from the cytosol, while the mitochondrial pool appears to be preserved; in hypoxia, at variance with the ischemic conditions, pyruvate does not show any beneficial effect. The action of pyruvate appears to be multifactorial and its effects are discussed by considering its action on the hydrogen peroxide breakdown, protection of pyruvate dehydrogenase, anaerobic production of ATP and diminution of the intracellular concentration of inorganic phosphate. PMID- 8232248 TI - Molecular and biochemical features of poly (ADP-ribose) metabolism. AB - In the past five years, poly(ADP-ribosyl)ation has developed greatly with the help of molecular biology and the improvement of biochemical techniques. In this article, we describe the physico-chemical properties of the enzymes responsible for the synthesis and degradation of poly(ADP-ribose), respectively poly(ADP ribose) polymerase and poly(ADP-ribose) glycohydrolase. We then discuss the possible roles of this polymer in DNA repair and replication as well as in cellular differentiation and transformation. Finally, we put forward various hypotheses in order to better define the function of this polymer found only in eucaryotes. PMID- 8232250 TI - Fatty acid-binding protein and its relation to fatty acid oxidation. AB - A relation between fatty acid oxidation capacity and cytosolic FABP content was found in heart and various muscles of the rat. Other tissues do not show such a relation, since they are involved in more or other pathways of fatty acid metabolism. At postnatal development FABP content and fatty acid oxidation capacity rise concomitantly in heart and quadriceps muscle in contrast to in liver and kidney. A dietary fat content of 40 en.% increased only the FABP content of liver and adipose tissue. Peroxisomal proliferators increased fatty acid oxidation in both liver and kidney, but only the FABP content of liver, and had no effect on heart and skeletal muscle. The FABP content of muscle did not show adaptation to various conditions. Only it increased in fast-twitch muscles upon chronic electrostimulation and endurance training. PMID- 8232251 TI - Modulation of fatty acid-binding protein content of adult rat heart in response to chronic changes in plasma lipid levels. AB - The aim of this work was to study in the adult rat heart the effect of modifications of fatty acid (FA) supply on the content of cytoplasmic fatty acid binding protein (H-FABPc). To modify the amount of circulating lipids, three different treatments were chosen: (i) an hypolipidemic treatment with Clofibrate, administered daily through a gastric tube at a dose of 250 mg/kg per day for one week, (ii) a continuous intravenous infusion of 20% Intralipid, a fat emulsion, for one week at a dose of 96 ml/kg per day, and (iii) a normobaric hypoxia exposure (pO2 = 10%) for three weeks. At the end of each treatment plasma lipids, myocardial H-FABPc content and the activities of three key enzymes (citrate synthase, CS, fructose-6-phosphate kinase, FPK and hydroxy-acyl CoA dehydrogenase, HAD) were assessed. With each of the three treatments a decrease of plasma cholesterol and phospholipid levels was observed. Plasma FA concentration increased with Intralipid infusion and decreased with chronic hypoxia. The heart H-FABPc content was increased by 20% with Clofibrate, decreased by 20% with chronic hypoxia and remained unaltered upon Intralipid treatment. The induced changes in H-FABPc content were not related directly to changes in plasma lipid levels. CS activity was slightly decreased in the hypoxia group, FPK activity decreased in the Clofibrate group, and HAD activity decreased in the Intralipid group. Among the various groups heart H-FABPc content was related to HAD activity. In conclusion, the H-FABPc content of adult rat heart appears responsive to changes in plasma lipid levels. PMID- 8232252 TI - Expression and localization of intestinal 15 kDa protein in the rat. AB - Rat intestinal 15 kDa protein (I-15P) is highly homologous to porcine gastrotropin. We studied the occurrence, distribution and subcellular localization of I-15P in the entire rat body, using the immunocytochemistry to localize protein and in situ hybridization to localize mRNA. Both techniques demonstrated the expression of I-15P in the enterocytes of ileum, luteal cells of ovary and a subpopulation of steroid-endocrine cells of adrenal gland. Immuno electron microscopy further demonstrated that I-15P is localized in both the cytoplasmic and nuclear matrix regions of these cells. The present results suggest roles of I-15P not only in the transport of bile salts but also in the metabolisms of certain steroid hormones. PMID- 8232253 TI - Retinal FABP principally localizes to neurons and not to glial cells. AB - The presence of fatty acid-binding protein (FABP) in the embryonic chick retina may be linked to the demand for polyunsaturated fatty acids in this developing neural tissue. There is a decline in the overall level of FABP as the retina matures, suggesting a role for FABP in cellular differentiation. However, this pattern is not present in the chick brain, indicating a unique function for FABP in the retina. Immunohistochemical staining of paraffin sections of chick retina from embryonic day 21 revealed immunopositive photoreceptor inner segments, outer nuclear layer, 'radial processes' in the inner nuclear layer, a subpopulation of cells in the ganglion cell layer, and inner limiting membrane. This pattern suggested that FABP positive cells were photoreceptors, Muller (glial) cells, and possibly ganglion cells. Staining of sections for glutamine synthetase, an enzyme specific for Muller cells, was similar but not identical to the pattern observed with FABP; thus identification of these cells as FABP-positive was not conclusive. However, in retinal cells dissociated from day E14 embryos and cultured for one week, staining with FABP was more intense in the neurons than in the 'flat' cells (presumed to be derived from the Muller cells). Retinal FABP thus appears to be localized predominantly in neurons, and may serve to sequester fatty acids in preparation for neurite outgrowth as the retinal cells differentiate. PMID- 8232254 TI - The function of acyl-CoA-binding protein (ACBP)/diazepam binding inhibitor (DBI). AB - Acyl-CoA-binding protein has been isolated independently by five different groups based on its ability to (1) displace diazepam from the GABAA receptor, (2) affect cell growth, (3) induce medium-chain acyl-CoA-ester synthesis, (4) stimulate steroid hormone synthesis, and (5) affect glucose-induced insulin secretion. In this survey evidence is presented to show that ACBP is able to act as an intracellular acyl-CoA transporter and acyl-CoA pool former. The rat ACBP genomic gene consists of 4 exons and is actively expressed in all tissues tested with highest concentration being found in liver. ACBP consists of 86 amino acid residues and contains 4 alpha-helices which are folded into a boomerang type of structure with alpha-helices 1, 2 and 4 in the one arm and alpha-helix 3 and an open loop in the other arm of the boomerang. ACBP is able to stimulate mitochondrial acyl-CoA synthetase by removing acyl-CoA esters from the enzyme. ACBP is also able to desorb acyl-CoA esters from immobilized membranes and transport and deliver these for mitochondrial beta-oxidation. ACBP efficiently protects acetyl-CoA carboxylase and the mitochondrial ADP/ATP translocase against acyl-CoA inhibition. Finally, ACBP is shown to be able to act as an intracellular acyl-CoA pool former by overexpression in yeast. The possible role of ACBP in lipid metabolism is discussed. PMID- 8232255 TI - Purification and partial characterisation of an alpha-tocopherol-binding protein from rabbit heart cytosol. AB - An alpha-tocopherol-binding protein has been isolated and purified from rabbit heart cytosol. The purified protein had an apparent molecular mass of 14,200, as derived from SDS-PAGE. The content of the protein in rabbit heart was around 11.8 micrograms per g of tissue. The binding of alpha-tocopherol to the purified protein was rapid, reversible, and saturable. Neither gamma nor delta tocopherol could displace the bound alpha-tocopherol from the protein, suggesting a high specificity for alpha-tocopherol. alpha-Tocopherol-binding protein did not bind oleate. Transfer of alpha-tocopherol from liposomes to mitochondria was stimulated 8-fold in the presence of the binding protein, suggesting that this protein may be involved in the intracellular transport of alpha-tocopherol in the heart. PMID- 8232256 TI - Role of fatty acid-binding protein in lipid metabolism of insect flight muscle. AB - Since insect flight muscles are among the most active muscles in nature, their extremely high rates of fuel supply and oxidation pose interesting physiological problems. Long-distance flights of species like locusts and hawkmoths are fueled through fatty acid oxidation. The lipid substrate is transported as diacylglycerol in the blood, employing a unique and efficient lipoprotein shuttle system. Following diacyglycerol hydrolysis by a flight muscle lipoprotein lipase, the liberated fatty acids are ultimately oxidized in the mitochondria. Locusta flight muscle cytoplasm contains an abundant fatty acid-binding protein (FABP). The flight muscle FABP of Locusta migratoria is a 15 kDa protein with an isoelectric point of 5.8, binding fatty acids in a 1:1 molar stoichiometric ratio. Binding affinity of the FABP for long-chain fatty acids (apparent dissociation constant Kd = 5.21 +/- 0.16 microM) is however markedly lower than that of mammalian FABPs. The NH2-terminal amino acid sequence shares structural homologies with two insect FABPs recently purified from hawkmoth midgut, as well as with mammalian FABPs. In contrast to all other isolated FABPs, the NH2 terminus of locust flight muscle FABP appeared not to be acetylated. During development of the insect, a marked increase in fatty acid binding capacity of flight muscle homogenate was measured, along with similar increases in both fatty acid oxidation capacity and citrate synthase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232257 TI - Solution structure of bovine heart fatty acid-binding protein (H-FABPc). AB - Fatty acid-binding protein (FABP) from bovine heart, a 15 kDa cytoplasmic protein has been investigated by multi-dimensional homonuclear and heteronuclear NMR spectroscopy. Perdeuterated palmitic acid has been used as fatty acid ligand. The tertiary structure has been determined from distance geometry calculations with the variable target functions algorithm (DIANA) utilizing 1027 interproton distance constraints, which were obtained from 1H-homonuclear NOESY spectra. Overlapping NOE crosspeaks were assigned by heteronuclear multidimensional NMR experiments with a 15N-labelled sample. The tertiary structure resembles a beta barrel (beta-clam) consisting of ten anti-parallel beta-strands and a short helix turn-helix motif. The beta-strands are arranged in two nearly orthogonal beta sheets composed of 5 strands each. The solution structure is compared with the x ray crystal structure of bovine heart and rat intestinal FABPs. PMID- 8232258 TI - Developmental changes of FABP concentration, expression, and intracellular distribution in locust flight muscle. AB - M-FABP from flight muscle of the locust, Schistocerca gregaria, is similar to mammalian H-FABP in its physical characteristics and amino acid sequence. We have studied developmental changes using ELISA, Northern Blotting, and EM/immuno-gold techniques. M-FABP is found in cytoplasm and nuclei, but not in mitochondria. It is the most abundant soluble muscle protein in fully developed adult locusts, comprising 18% of the total cytosolic protein. However, no FABP is detectable at the beginning of the adult stage. Its concentration rises dramatically during the next 10 days, after which it reaches its maximal value. Expression apparently is turned on after ecdysis and continues for 10 days; thereafter, FABP mRNA diminishes and reaches a constant, but low level, probably needed to maintain the current FABP level. From a series of experiments employing metamorphosis controlling hormones and antihormones it is evident that the induction of FABP expression is directly linked to metamorphosis. PMID- 8232259 TI - Characterization of the non-specific lipid transfer protein EP2 from carrot (Daucus carota L.). AB - The extracellular protein EP2 was previously identified as non-specific lipid transfer protein based on its cDNA-derived amino acid sequence. Here, the purification of the EP2 protein from the medium of somatic embryo cultures is described. After two cycles of ion-exchange and gel permeation chromatography, a single silver-stained protein band with an apparent molecular mass of 10 kDa was observed on SDS-PAGE. This protein band was recognized by the antiserum raised against a EP2-beta-galactosidase fusion-protein. Employing a fluorescent phospholipid analog, it was shown that the purified EP2 protein is capable of binding phospholipids and is able to enhance their transfer between artificial membranes. Employing a gel permeation assay, it could be demonstrated that the EP2 protein is also capable of binding palmitic and oleic acid as well as oleyl CoA. Because in plants these fatty acids are used as precursor molecules for cutin, these results are in support of the proposed role of the EP2 protein to transport cutin monomers from their site of synthesis through the cell wall of epidermal cells to sites of cutin polymerization. PMID- 8232260 TI - Significance of cytoplasmic fatty acid-binding protein for the ischemic heart. AB - Ischemia of the heart is accompanied by the tissue accumulation of long-chain fatty acids and their metabolic derivatives such as beta-hydroxy fatty acids and fatty acyl-CoA and acyl-L-carnitine esters. These substances might be detrimental for proper myocardial function. Previously, it has been suggested that intracellular lipid binding proteins like cytoplasmic fatty acid-binding protein (FABP) and acyl-CoA binding protein (ACBP) may bind these accumulating fatty acyl moieties to prevent their elevated levels from potentially harmful actions. In addition, the suggestion has been made that the abundantly present FABP may scavenge free radicals which are generated during reperfusion of the ischemic heart. However, these protective actions are challenged by the continuous physico chemical partition of fatty acyl moieties between FABP and membrane structures and by the rapid release of FABP from ischemic and reperfused cardiac muscle. Careful evaluation of the available literature data reveals that at present no definite conclusion can be drawn about the potential protective effect of FABP on the ischemic and reperfused heart. PMID- 8232261 TI - Release of fatty acid-binding protein and long chain fatty acids from isolated rat heart after ischemia and subsequent calcium paradox. AB - To obtain insight into the relation between the release of heart-type fatty acid binding protein (H-FABPc) and of long-chain fatty acids (FA) from injured cardiac tissue, rat hearts were Langendorff perfused according to the following scheme: 30 min normoxia, 60 min ischemia, 30 min reperfusion, 10 min Ca2+ free perfusion and finally 10 min Ca2+ repletion. During this protocol right ventricular (Qrv) and interstitial effluent samples (Qi) were collected at regular intervals. During reperfusion a total of 0.8 +/- 0.1 nmol H-FABPc but no FA were detected in the effluents. However, during Ca2+ readmission, 45 +/- 4 nmol H-FABPc (80-90% of total tissue content) was released with an initial (first 3 min) simultaneous release of FA (FA/H-FABPc ratio 0.90 +/- 0.07 mol/mol). Thereafter, FA release continued at 10-15 nmol per min mainly in Qrv while the rate of H-FABPc release decreased. During Ca2+ repletion, tissue FA content raised rapidly from 168 +/- 20 to 1918 +/- 107 nmol/g dry weight. These findings suggest that after severe cardiac damage initially FA is released bound to H-FABPc, whereas further FA release occurs in a non-protein bound manner. PMID- 8232262 TI - Fatty acid-binding proteinuria diagnoses myocardial infarction in the rat. AB - Cytoplasmic heart-type fatty acid-binding protein (H-FABPc) is a low molecular weight protein with abundant presence in the myocardium. Upon ischemia it is released from the heart and can subsequently be detected in plasma and urine. In this study, the value of measurement of H-FABPc excreted into urine for the diagnosis of myocardial infarction (MI) is investigated in the rat. To this end, firstly the kinetic behaviour of H-FABPc in plasma was examined and its release into urine quantified. After injection of purified H-FABPc in normal animals, a net recovery in urine of 14-29% was found. The kinetic behaviour of H-FABPc in plasma was characterized by a total clearance of 0.33 ml/min and a half-life value of total elimination of about 270 min. Knowing these plasma characteristics of H-FABPc, a comparison was made between the cumulative amounts of H-FABPc released in the 24-hour urine of MI rats and of sham-operated animals. In MI rats, with a mean morphometric MI size of 43%, the mean total amount of H-FABPc excreted into urine was 79 micrograms, while in sham-operated rats this was 23 micrograms. This difference between both groups is significant (p < 0.001). It is concluded that urinary H-FABPc can be used as a noninvasive marker for MI in the rat. PMID- 8232263 TI - Cellular binding proteins for fatty acids and retinoids: similar or specialized functions? AB - The cellular fatty acid-binding proteins (FABP) and cellular retinoid (retinol, retinoic acid)-binding proteins (CRtBP) are structurally and functionally-defined groups within an evolutionarily conserved gene family. CRtBP are expressed in both fully differentiated and developing tissues in a manner that supports a relationship to the action of retinoic acid in morphogenesis and cellular differentiation. The FABP are, by contrast, expressed only in fully differentiated tissues in a manner compatible with a major function in the metabolism of long-chain fatty acids (LCFA) for energy production or storage. The precise function(s) of FABP and CRtBP remain imperfectly understood, while subspecialization of function(s) within the two groups is suggested by the complex diversity in both of structurally distinct members that display striking tissue and temporal specificity of expression in addition to ligand specificity. Notwithstanding this considerable apparent functional diversity among the FABP and CRtBP, available evidence supports a dual set of generic functions for both protein groups in a) promoting cellular flux of poorly water-soluble ligands and their subsequent metabolic utilization or transformation, and b) sequestration of ligands in a manner that limits their association with alternative binding sites within the cell, of which members of the steroid hormone nuclear receptor superfamily (HNR) are a potentially important category. Theoretical as well as experimental models probing diffusional fluxes of LCFA in vitro and in living cells have provided support for a function for FABP in intracellular LCFA transport. Protein-bound ligand also appears to provide the substrate for metabolic transformation of retinoids bound to CRtBP, but convincing evidence is lacking for an analogous mechanism in the direct facilitation of fatty acid utilization by FABP. An emerging relationship between FABP and CRtBP function centers on their binding of, and induction by, ligands which activate or transform specific HNR-the retinoic acid receptors and the peroxisome proliferator activated receptor in the case of CRtBP and FABP, respectively. Evidence consistent with both a 'promotive' role (provision of ligands for HNR) and a 'protective' role (limiting availability of free ligand for HNR association) has been advanced for CRtBP. Available data supports a 'protective' function for cellular retinoic acid-binding proteins (CRABP) and liver FABP (L FABP) and points to the existence of ligand-defined, lipid-binding-protein-HNR relationships in which CRABP serve to attenuate the induction of gene expression by retinoic acid, and in which L-FABP may modulate a cellular adaptive multigene response to increased LCFA flux or compromised LCFA utilization.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8232264 TI - Involvement of arginine in the binding of heme and fatty acids to fatty acid binding protein from bovine liver. AB - Fatty acid-binding protein from bovine liver but not from bovine heart binds hematin in a saturable manner with high affinity. This property is not confined to a particular isoform as both, pI 6.0- and pI 7.0 L-FABP, bind hematin similarly. In competition experiments hematin and oleic acid could replace each other demonstrating that they share at least parts of the same binding site. Common structural features, i.e. the presence of carboxylic groups and of hydrophobic carbon chains led to the hypothesis that both ligands interact similarly with L-FABP. This was supported by the decrease of binding affinity for either ligand upon modification with phenylglyoxal. Modification in the presence of fatty acid revealed the protection of one of the two arginines of L-FABP. By peptide mapping and Edman degradation Arg122 was identified as the counterpart of the fatty acids carboxylic group. PMID- 8232265 TI - Titration calorimetry as a binding assay for lipid-binding proteins. AB - Titration calorimetry has been evaluated as a method for obtaining binding constants and thermodynamic parameters for the cytosolic fatty acid- and lipid binding proteins. An important feature of this method was its ability to accurately determine binding constants in a non-perturbing manner. The equilibrium was not perturbed, since there was no requirement to separate bound and free ligand in order to obtain binding parameters. Also, the structure of the lipid-protein complex was not perturbed, since native ligands were used rather than non-native analogues. As illustrated for liver fatty acid-binding protein, the method distinguished affinity classes whose dissociation constants differed by an order of magnitude or less. It also distinguished endothermic from exothermic binding reactions, as illustrated for the binding of two closely related bile salts to ileal lipid-binding protein. The main limitations of the method were its relatively low sensitivity and the difficulty working with highly insoluble ligands, such as cholesterol or saturated long-chain fatty acids. However, the signal-to-noise ratio was improved by manipulating the buffer conditions, as illustrated for oleate binding to rat intestinal fatty acid binding protein. Binding parameters are reported for oleate interactions with several wild-type and mutant lipid-binding proteins from intestine. Where possible, the binding parameters obtained from calorimetry were compared with results obtained from fluorescence and Lipidex binding assays of comparable systems. PMID- 8232266 TI - High resolution X-ray studies of mammalian intestinal and muscle fatty acid binding proteins provide an opportunity for defining the chemical nature of fatty acid: protein interactions. AB - The structure of E. coli-derived rat intestinal fatty acid-binding protein has recently been refined to 1.2 A without bound fatty acid and to 2.0 A and 1.75 A with bound hexadecanoate (palmitate) and 9Z-octadecenoate (oleate), respectively. The structure of E. coli-derived human muscle fatty acid-binding protein has also been solved to 2.1 A with a C16 bacterial fatty acid. Both proteins contain 10 anti-parallel beta-strands in a +1, +1, +1... motif. The strands are arranged in two beta-pleated sheets that are orthogonally oriented. In each case, the fatty acid is enclosed by the beta-sheets and is bound to the proteins by feeble forces. These feeble forces consist of (i) a hydrogen bonding network between the fatty acid's carboxylate group, ordered solvent, and side chains of polar/ionizable amino acid residues; (ii) van der Waals contacts between the methylene chain of the fatty acid and the side chain atoms of hydrophobic and aromatic residues; (iii) van der Waals interactions between the omega-terminal methyl and the component methenyls of the phenyl side chain of a Phe which serves as an adjustable terminal sensor situated over a surface opening or portal connecting interior and exterior solvent; and (iv) van der Waals contacts between methylenes of the alkyl chain and oxygens of ordered waters that have been located inside the binding cavity. These waters are positioned over one face of the ligand and are held in place by hydrogen bonding with one another and with the side chains of protein's polar and ionizable residues. Binding of the fatty acid ligand is associated with minimal adjustments of the positions of main chain or side chain atoms. However, acquisition of ligand is associated with removal of ordered interior solvent suggesting that the free energy of dehydration of the binding site may be as important for the energy of the binding reaction as the free energy of stabilization of the fatty acid: protein complex. PMID- 8232267 TI - Identification of high affinity membrane-bound fatty acid-binding proteins using a photoreactive fatty acid. AB - A photoaffinity labeling method was developed to identify and characterize high affinity fatty acid-binding proteins in membranes. The specific labeling of these sites requires the use of low concentrations (nanomolar) of the photoreactive fatty acid 11-m-diazirinophenoxy-[11-3H]undecanoate. It was delivered as a bovine serum albumin (BSA) complex which serves as a reservoir for fatty acid and thus allows precise control of unbound fatty acid concentrations. The fadL protein of E. coli, which is required for fatty acid permeation of its outer membrane, was labeled by the photoreactive fatty acid neither specifically nor saturably when the probe was added in the absence of BSA; however when a nanomolar concentration of the uncomplexed probe was maintained in the presence of BSA, the labeling of the fadL protein was highly specific and saturable. This photoaffinity labeling method was also used to characterize a 22 kDa, high affinity fatty acid-binding protein which we have recently identified in the plasma membrane of 3T3-L1 adipocytes. This protein bound the probe with a Kd of 216 nM. The approach described is easily capable of identifying membrane-bound fatty acid-binding proteins and can distinguish between those of high and low affinities for fatty acids. It represents a general method for the identification and characterization of fatty acid-binding proteins. PMID- 8232268 TI - Diversity of fatty acid-binding protein structure and function: studies with fluorescent ligands. AB - The mammalian fatty acid-binding proteins (FABP) are localized in many distinct cell types. They bind long chain fatty acids in vitro, however, their functions and mechanisms of action in vivo remain unknown. The present studies have sought to understand the relationships among these proteins, and to address the possible role of FABP in cellular fatty acid traffic. A series of anthroyloxy-labeled fluorescent fatty acids have been used to examine the physicochemical properties of the fatty acid-binding sites of different members of the FABP family. The fatty acid probes have also been used to study the rate and mechanism of fatty acid transfer from different FABP types to phospholipid membranes. The results of these studies show a number of interesting and potentially important differences between FABP family members. An examination of adipocyte and heart FABP (A- and H FABP) shows that their fatty acid-binding sites are less hydrophobic than the liver FABP (L-FABP) site, and that the bound ligand experiences less motional constraint within the A- and H-FABP binding sites than within the L-FABP binding site. In keeping with these differences in structural properties, it was found that anthroyloxy-fatty acid transfer from A- and H-FABP to membranes is markedly faster than from L-FABP. Moreover, the mechanism of fatty acid transfer was found to be similar for the highly homologous A- and H-FABP, whereby transfer to phospholipid membranes appears to occur via transient collisional interactions between the FABP and membranes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232269 TI - Genome organization and expression of the rat ACBP gene family. AB - Acyl-CoA-Binding Protein (ACBP)/Diazepam-Binding Inhibitor (DBI) is a 10 kD protein which has been implicated in a surprisingly large number of biochemical functions. We have unambiguously demonstrated that ACBP binds acyl-CoA esters with high affinity and in vivo functions as an acyl-CoA ester pool former. We have molecularly cloned and characterized the rat ACBP gene family which comprises one expressed and four processed pseudogenes. One of these was shown to exist in two allelic forms. A comprehensive computer-aided analysis of the promoter region of the expressed ACBP gene revealed that it exhibits all the hallmarks of typical housekeeping genes. In addition, the promoter region harbors a number of potential tissue specific cis-acting elements that may in part regulate the level of ACBP expression in specialized cells. PMID- 8232270 TI - Expression of rat L-FABP in mouse fibroblasts: role in fat absorption. AB - Fatty acid-binding proteins (FABP) are abundant cytosolic proteins whose levels is responsive to nutritional, endocrine, and a variety of pathological states. Although FABPs have been investigated in vitro for several decades, little is known of their physiological function. Liver L-FABP binds both fatty acids and cholesterol. Competitive binding analysis and molecular modeling studies of L FABP indicate the presence of two ligand binding pockets that accommodate one fatty acid each. One fatty acid binding site is identical to the cholesterol binding site. To test whether these observations obtained in vitro were physiologically relevant, the cDNA encoding L-FABP was transfected into L-cells, a cell line with very low endogenous FABP and sterol carrier proteins. Uptake of both ligands did not differ between control cells and low expression clones. In contrast, both fatty acid uptake and cholesterol uptake were stimulated in the high expression cells. In high expression cells, uptake of fluorescent cis parinaric acid was enhanced more than that of trans-parinaric acid. This is consistent with the preferential binding of cis-fatty acids to L-FABP but in contrast to the preferential binding of trans-parinaric acid to the L-cell plasma membrane fatty acid transporter (PMFABP). These data show that the level of cytosolic fatty acids in intact cells can regulate both the extent and specificity of fatty acid uptake. Last, sphingomyelinase treatment of L-cells released cholesterol from the plasma membrane to the cytoplasm and stimulated microsomal acyl-CoA: cholesteryl acyl transferase (ACAT). This process was accelerated in high expression cells. These observations show for the first time in intact cells that L-FABP, a protein most prevalent in liver and intestine where much fat absorption takes place, may have a role in fatty acid and cholesterol absorption. PMID- 8232271 TI - Research of an in vitro model to study the expression of fatty acid-binding proteins in the small intestine. AB - In order to find an in vitro model for studying the regulation of the biosynthesis of the cytoplasmic Fatty Acid-Binding Proteins (FABPc) expressed in the small intestine, Intestinal- and Liver- (I- and L-) FABPc expressions were tested by Northern blotting in 8 normal or cancerous intestinal cell lines from man, mouse and rat and in organ culture of mouse jejunal explants. Neither I- nor L-FABPc mRNA was detected in any cell strains tested except in the highly differentiated human enterocyte-like intestinal cell line Caco-2. In this line, Northern blot analysis revealed a single messenger of about 0.7 kb corresponding to the L-FABPc. A two-fold increase in mRNA L-FABPc occurred in differentiated Caco-2 cells treated for 7 days with 0.05 mM bezafibrate, a peroxisome proliferating hypolipidemic drug. The lack of I-FABPc messengers in this strain led us to seek another in vitro model. I- and L-FABPc messengers were found using an organ culture of mouse jejunal explants. A clear rise in I- and, especially, L FABPc mRNA levels occurred 6 and 24 hr after the addition of 0.05 mM bezafibrate in the culture medium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232272 TI - Mechanisms of regulation of liver fatty acid-binding protein. AB - Liver fatty acid-binding protein (L-FABP) expression is modulated by developmental, hormonal, dietary, and pharmacological factors. The most pronounced induction is seen after treatment with peroxisome proliferators, which induce L-FABP coordinately with microsomal cytochrome P-450 4A1 and the enzymes of peroxisomal fatty acid beta-oxidation. These effects of peroxisome proliferators may be mediated by a receptor which has been shown to be activated by peroxisome proliferators in mammalian cell transfection studies. However, the peroxisome proliferators tested thus far do not bind to this receptor, known as the peroxisome proliferator-activated receptor (PPAR), and its endogenous ligand(s) also remain unknown. Peroxisome proliferators inhibit mitochondrial beta-oxidation, and one hypothesis is that the dicarboxylic fatty acid metabolites of accumulated LCFA, formed via the P-450 4A1 omega-oxidation pathway, serve as primary inducers of L-FABP and peroxisomal beta-oxidation. We have tested this hypothesis in primary hepatocyte cultures exposed to clofibrate (CF). Inhibition of P-450 4A1 markedly diminished, via a pre-translational mechanism, the CF induction of L-FABP and peroxisomal beta-oxidation. In further experiments, long-chain dicarboxylic acids, the final products of the P-450 4A1 omega-oxidation pathway, but not LCFA, induced L-FABP and peroxisomal beta oxidation pre-translationally. These results suggest a role, in part, for long chain dicarboxylic acids in mediating the peroxisome proliferator induction of L FABP and peroxisomal beta-oxidation. We also found that LCFA, which undergo rapid hepatocellular metabolism, could become inducers of L-FABP and peroxisomal beta oxidation under conditions where their metabolism was inhibited.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232273 TI - Effect of spermine on membrane-associated and membrane-inserted forms of protein kinase C. AB - Protein kinase C is reported to exist in two membrane-bound states: a reversible one which can be dissociated by calcium chelators (membrane-associated form) and an irreversible one which is chelator stable (membrane-inserted form). In the present work the effects of a naturally occurring polyamine (spermine) on the membrane-associated and membrane-inserted forms of protein kinase C were investigated using a reconstituted system consisting of partially purified protein kinase C from rat brain and phospholipid vesicles of defined composition. The active membrane-bound complex was conveniently determined by its ability to bind radioactive phorbol ester with an exact 1:1 stoichiometry. Our experimental data show that, in the absence of calcium ions, the amount of enzyme bound to phospholipids vesicles was dramatically reduced by the presence of spermine whereas the PDBu binding affinity was not significantly affected. The addition of the divalent cation increased the affinity of phorbol ester for the active complex but had no effect on Nmax; spermine added in this experimental conditions was no longer able to decrease the total number of enzyme molecules bound to liposomes. Moreover gel filtration experiments of the protein kinase C phospholipids complex formed in the presence of calcium, indicated that polyamine added during the association process was able to reduce the extent of enzyme insertion into liposomes. Since the increase in phospholipid concentration resulted in a higher level of non-dissociable protein kinase C-liposomes complex we propose that spermine, complexing to membrane binding sites both in the absence and in the presence of Ca++, could promote binding conditions that oppose to the formation of the inserted form of the enzyme. As a consequence the distribution between the reversible and the irreversible membrane-bound forms of protein kinase C is affected. PMID- 8232274 TI - Distribution and regulation of natriuretic factor-R1C receptor subtypes in mammalian cell lines. AB - The differential distribution of natriuretic peptide receptor subtypes and their distinct properties were assessed in mammalian cellular models which were screened for their ability to produce cGMP upon stimulation by different natriuretic peptides. The ANF-R1A receptor subtype was distinguished by its selective activation by atrial natriuretic factor (ANF) while the ANF-R1C was characterized by preferential stimulation by C-type natriuretic peptide (CNP). AT 620 pituitary cells, bovine adrenal chromaffin cells, and NIH-3T3 fibroblasts mainly express the ANF-R1C receptor subtype. Other cell lines such as PC12, RASM and GH3 express significant but varying amounts of both ANF-R1A and ANF-R1C subtypes. A10 and NIH cells which express high density of ANF-R2 receptor subtype, also demonstrate a higher sensitivity to CNP over ANF suggesting that they express significant amounts of ANF-R1C. Studies of the regulation by ATP of guanylyl cyclase activity indicate that both ANF-R1A and ANF-R1C subtypes are modulated in the same manner. In the presence of Mn2+, ATP inhibits the CNP stimulated guanylyl cyclase activity while in the presence of Mg2+ adenine nucleotides potentiate the stimulation by CNP. In addition, we show that like the ANF-R1A, the ANF-R1C guanylyl cyclase activity can be regulated by phosphorylation since preincubation with TPA or FKL attenuates the subsequent stimulation by CNP in cultured cells. The results presented demonstrate that specific cell types express distinct natriuretic peptide receptor subtypes and also that the newly characterized ANF-R1C subtype is regulated by ATP and serine/threonine kinases in the same way as the ANF-R1A subtype. PMID- 8232275 TI - Role of Ca(2+)-calmodulin dependent phospholamban phosphorylation on the relaxant effect of beta-adrenergic agonists. AB - The role of the Ca(2+)-calmodulin dependent pathway of phospholamban phosphorylation on the relaxant effect of beta-adrenergic agonists was studied in isolated perfused rat heart. Administration of the calmodulin antagonist W7 or lowering [Ca]o from 1.35 mM (control) to 0.25 mM, were used as experimental tools to inhibit the Ca(2+)-calmodulin dependent protein kinase activity. 3 x 10(-8) M isoproterenol increased cAMP levels from 0.613 +/- 0.109 pmol/mg wet weight to 1.581 +/- 0.123, phospholamban phosphorylation from 36 +/- 6 pmol 32P/mg protein to 277 +/- 26 and decreased time to half relaxation (t1/2) from 61 +/- 2 msec to 39 +/- 2. Simultaneous perfusion of isoproterenol with 10(-6) M W7, decreased phospholamban phosphorylation to 170 +/- 23 and prolongated t1/2 to 47 +/- 3 but did not affect the increase either in cAMP levels or myocardial contractility produced by isoproterenol. Similar effects on phospholamban phosphorylation and myocardial relaxation were obtained when isoproterenol was perfused in low [Ca]o. Low [Ca]o did not affect the increase in cAMP elicited by isoproterenol but offset the positive inotropic effect of the beta-agonist. The results suggest a physiological role of the Ca(2+)-calmodulin dependent phospholamban phosphorylation pathway as a mechanism that supports, in part, the beta adrenergic cardiac relaxant effect. PMID- 8232276 TI - S-adenosyl-l-methionine inhibits phosphoinositide metabolism in the rat brain synaptosomal suspensions. AB - S-adenosyl-l-methionine (AdoMet) has been reported to affect events linked to noradrenergic neurotransmission. In the present work, we studied the effect of AdoMet on norepinephrine (NE)-stimulated inositol phosphate production in 3H inositol-labelled crude synaptosomal suspensions of rat brain. AdoMet (50-1000 microM) decreased both the synthesis of labelled polyphosphoinositide (30-50%) and the release of inositol mono- and bisphosphate (40-50%). The AdoMet effect was not dependent on NE concentration (10-1000 microM), suggesting that the inhibition of inositol phosphate release was not the result of a modification of the norepinephrine binding to its receptor sites. S-adenosyl-L-homocysteine (AdoHcy) (1 mM) an inhibitor of methyltransferase activities, partially inhibited (70%) the AdoMet (0.1 mM) effect, indicating that the methylation processes cannot explain all the effects observed. We conclude that, in addition to previously reported effects of AdoMet on NE transport, AdoMet may reduce NE linked intracellular signalling. PMID- 8232277 TI - Oxygen free radical induced damage during intestinal ischemia/reperfusion in normal and xanthine oxidase deficient rats. AB - This study looks at the role of xanthine oxidase (XO) in ischemia/reperfusion (I/R) induced intestinal mucosal damage using normal and xanthine oxidase deficient rats. Tungstate feeding for 3 days depleted the intestinal mucosal XO by 80%. A ligated loop of the rat small intestine (both normal and XO-deficient) was subjected to 1 h of total ischemia followed by 5 min revascularisation. The ensuing mucosal damage was assessed by biochemical and histological studies. Ischemia or I/R increased the XO levels in normal rats without any change in XO deficient rats. Myeloperoxidase (a neutrophil marker) level was increased in both group of rats but it was comparatively higher in the XO-deficient rats. Accumulation of peroxidation products such as malondialdehyde, conjugated diene and increased production of hydroxyl radicals by microsomes were seen after ischemia and I/R and were similar in normal and XO-deficient rats. Studies on other parameters of peroxidation showed a decrease in polyunsaturated fatty acids and alpha-tocopherol, an increase in cysteine and cystine levels after I/R and were similar in both normal and XO-deficient rats. Histological results indicated gross morphological changes in the intestinal mucosa due to ischemia and I/R, and the damage was more severe in XO-deficient rats. These observations suggest that oxygen-derived free radicals are involved in the intestinal mucosal damage during I/R and infiltrated neutrophils rather than XO may be the primary source of free radicals under these conditions. PMID- 8232278 TI - Methylamine-treated low density lipoproteins elicit different responses in HepG2 cells and macrophages. AB - Recent results from this laboratory have demonstrated the existence of labile thiolester bonds in apolipoprotein B (ApoB). Thiolester bonds can be cleaved with nucleophiles such as methylamine, resulting in conformational change. The purpose of this study was to explore whether the cellular interactions would be altered after methylamine treatment of low density lipoproteins (LDL). Human hepatoma cells, HepG2, and human monocyte derived macrophages were used for these studies. Fresh LDL were incubated with methylamine under mild alkaline conditions under N2 and with preservatives for 24 h. The methylamine-treated LDL showed particle size and net charge identical to fresh native LDL. In addition, no oxidative modification of LDL occurred under the experimental conditions. The methylamine treated LDL were indistinguishable from native LDL in HepG2 cells as judged by binding, degradation, cholesterol accumulation and de novo sterol synthesis. However, methylamine-treated LDL caused an increased accumulation of cholesteryl esters in macrophages which was comparable to the accumulation caused by acetylated LDL. Dual color digital imaging fluorescence microscopy revealed no competition between acetylated and methylamine-treated LDL, suggesting that the excessive uptake of methylamine-treated LDL was not mediated by the 'scavenger' receptor. The increased accumulation of cholesteryl ester in macrophages also did not appear to stem from the classical LDL receptor. These results suggest that a new receptor binding domain is exposed due to the conformational change upon treatment of LDL with methylamine. PMID- 8232279 TI - Serial deletion constructs of human cardiac myosin heavy chain genes generated by PCR amplification. AB - Serial deletion constructs derived from the 5'-flanking regions of the human cardiac alpha- and beta-myosin heavy chain genes were generated by polymerase chain reaction (PCR) amplifications. Generation of different length chimeric constructs were based on the complete sequence of the human cardiac myosin heavy chain genes. The primers were synthesized with HindIII and BamH1 sites and were linked to any designed nucleotide of the 5' flanking sequence of the myosin heavy chain gene(s). Following the PCR amplification and the site-directed mutagenesis, the PCR products were verified by DNA sequencing and subsequently ligated to the chloramphenicol acetyltransferase (pBLCAT3) reporter gene which was restricted with Hind III and BamH1. Neonatal rat cardiocytes were used to assay the promotor activity (i.e. CAT activity) of different lengths of the chimeric constructs of the gene. PMID- 8232280 TI - Gene expression: surprises from the class III side. AB - Those genes which are transcribed by RNA polymerase III continue to give surprising results with respect to their cis-acting elements and transacting factors. As a result, a broader view of class III promoters has emerged and the internal promoters are not universal in classical polymerase III genes. The involvement of TFIID, TFIIA, a factor homologous to TFIIB and an RNA factor in class III gene transcription has further changed our thinking in regards to the mechanisms of transcription. PMID- 8232281 TI - Influence of some biological response modifiers on swelling of rat liver mitochondria in vitro. AB - In order to understand any involvement of altered calcium functions in peroxidative membrane damage, the effect of a few chemicals, known to modify specific biological responses involving calcium related functions on mitochondrial swelling in vitro was studied. Histamine caused swelling, whereas antihistamines reduced calcium induced swelling. Anti-inflammatory agents aspirin and indomethacin did not affect the initial rapid phase of swelling but reduced the swelling during the later phase. The uncouplers of oxidative phosphorylation and electron transport chain blockers such as dinitrophenol (DNP), antimycin-A and rotenone reduced swelling and the respiratory inhibitors KCN and sodium azide completely abolished it. Trifluoperazine, an anti-calmodulin agent did not influence the initial phase of calcium induced swelling but in the subsequent phase swelling was reduced. c-AMP as well as calcium ionophores, calcimycin and lasalocid acid, potentiated swelling. Thus agents capable of modulating calcium functions could influence the in vitro swelling of mitochondria. PMID- 8232282 TI - Inhibition of monoamine oxidase-A of rat brain by pyrethroids--an in vitro kinetic study. AB - An in vitro kinetic study on inhibition of the monoamine oxidase-A (MAO-A) of the rat brain by two pyrethroids, namely permethrin (PM) and cypermethrin (CPM), has shown that PM and CPM competitively inhibit MAO-A by altering both the Michaelis Menten constant (Km) and the maximum velocity (Vmax). Inhibitor constant values (Ki) indicated that CPM was a more effective inhibitor of MAO-A than PM. Both PM and CPM caused maximum inhibition of MAO-A at neutral pH. CPM significantly elevated the activation energy values of MAO-A as compared to those of PM. PMID- 8232283 TI - Hormonal regulation of sialic acid-binding (SAS) protein synthesis of rat uterus. AB - Sialic acid binding proteins (SAS) of rat uteri have been found in all three stages of the estrous cycle. To study the control of synthesis of these proteins two different animal models were used I-immature female rats (25 d) where the hormones estradiol (E2) and progesterone (P4) were given separately and together, and II-adult female rats where hormone treatment commenced 14 days after ovariectomy. The data indicated that E2 initiated the synthesis of SAS proteins in the immature animals, while P4 could inhibit SAS synthesis, either given alone or together with E2. However, prior priming of the rat with E2 and subsequent administration of P4 stimulated SAS protein synthesis. PMID- 8232284 TI - Intracellular pH in Schizosaccharomyces pombe--comparison with Saccharomyces cerevisiae. AB - We examined cytoplasmic pH regulation in Schizosaccharomyces pombe and Saccharomyces cerevisiae using pH-sensitive fluorescent dyes. Of several different fluorescent compounds tested, carboxy-seminaphthorhodafluor-1 (C.SNARF 1) was the most effective. Leakage of C.SNARF-1 from S. pombe was much slower than leakage from C. cerevisiae. Using the pH-dependent fluorescence of C.SNARF-1 we showed that at an external pH of 7, mean resting internal pH was 7.0 for S. pombe and 6.6 for S. cerevisiae. We found that internal pH in S. pombe was maintained over a much narrower range in response to changes in external pH, especially at acidic pH. The addition of external glucose caused an intracellular alkalinization in both species, although the effect was much greater in S. cerevisiae than in S. pombe. The plasma membrane H(+)-ATPase inhibitor diethylstilbestrol reduced both the rate and extent of alkalinisation, with an IC50 of approximately 35 microM in both species. Amiloride also inhibited internal alkalinisation with IC50's of 745 microM for S. cerevisiae and 490 microM for S. pombe. PMID- 8232285 TI - Diethylglyoxal bis(guanylhydrazone), a potent inhibitor of mammalian S adenosylmethionine decarboxylase. Effects on cell proliferation and polyamine metabolism in L1210 leukemia cells. AB - The polyamines are cell constituents essential for growth and differentiation. S Adenosylmethionine decarboxylase (AdoMetDC) catalyzes a key step in the polyamine biosynthetic pathway. Methylglyoxal bis(guanylhydrazone) (MGBG) is an anti leukemic agent with a strong inhibitory effect against AdoMetDC. However, the lack of specificity limits the usefulness of MGBG. In the present report we have used an analog of MGBG, diethylglyoxal bis(guanylhydrazone) (DEGBG), with a much greater specificity and potency against AdoMetDC, to investigate the effects of AdoMetDC inhibition on cell proliferation and polyamine metabolism in mouse L1210 leukemia cells. DEGBG was shown to effectively inhibit AdoMetDC activity in exponentially growing L1210 cells. The inhibition of AdoMetDC was reflected in a marked decrease in the cellular concentrations of spermidine and spermine. The concentration of putrescine, on the other hand, was greatly increased. Treatment with DEGBG resulted in a compensatory increase in the synthesis of AdoMetDC demonstrating an efficient feedback control. Cells seeded in the presence of DEGBG ceased to grow after a lag period of 1-2 days, indicating that the cells contained an excess of polyamines which were sufficient for one or two cell cycles in the absence of polyamine synthesis. The present results indicate that analogs of MGBG, having a greater specificity against AdoMetDC, might be valuable for studies concerning polyamines and cell proliferation. PMID- 8232286 TI - In vitro oxidative inactivation of glutathione S-transferase from a freeze tolerant reptile. AB - We have previously reported that when garter snakes. Thamnophis sirtalis parietalis, a freeze tolerant species, were exposed to 5 h freezing at -2.5 degrees C organs showed increases in the activities of anti-oxidant enzymes, especially catalase in skeletal muscle. This was interpreted to be an adaptation to deal with the potentially injurious postischemic situation of thawing. The present work analyzes in vitro oxidative inactivation of a possible target of postischemic-induced free radical damage, the secondary anti-oxidant defense glutathione-S transferase, and the protective role of endogenous catalase. Approximately 50% of GST activity from snake muscle homogenates was lost within 2 min after addition of H2O2 plus Fe(II) (0.4-2 mM) in media containing azide whereas addition of iron alone resulted in no damaging effects. The opposing effects of dimethyl sulfoxide and EDTA in modifying this process strongly suggested the involvement of .OH radicals in the GST inactivation. A partial recovery of the activity was promoted by mercaptoethanol, indicating that sulphydryl groups oxidation participate in the mechanism of GST inactivation. Pre incubation of the reaction media containing H2O2 caused protection of the GST activity only in the absence of azide, indicating that endogenous catalase modulates the extent of oxyradical damage. The protective pre-incubation effect was more efficacious when employing homogenates from lung and liver, organs that have higher catalase activities, as well as homogenates from freezing-exposed muscle (that show an 80% increase in catalase activity, compared with control). The protection against GST inactivation observed in muscle from frozen snakes demonstrates that increased anti-oxidant defenses during freezing exposure can be a key factor in controlling in vitro oxyradical damage. The implications for natural freeze tolerance are discussed. PMID- 8232287 TI - Regulatory effect of regucalcin on (Ca(2+)-Mg2+)-ATPase in rat liver plasma membranes: comparison with the activation by Mn2+ and Co2+. AB - The effect of various metals and regucalcin, a calcium-binding protein isolated from rat liver cytosol, on (Ca(2+)-Mg2+)-ATPase activity in the plasma membranes of rat liver was investigated. Of various metals (Zn2+, Cu2+, Ni2+, Mn2+, Co2+ and Al3+; 100 microM as a final concentration), Mn2+ and Co2+ increased markedly (Ca(2+)-Mg2+)-ATPase activity, while other metals had no effect. When Ca2+ was not added into enzyme reaction mixture, Mn2+ and Co2+ (25-100 microM) did not significantly increase the enzyme activity, indicating that heavy metals act on Ca(2+)-stimulated phosphorylation of the enzyme. Meanwhile, regucalcin (0.25-1.0 microM) caused a remarkable elevation of (Ca(2+)-Mg2+)-ATPase activity. This increase was not inhibited by the presence of 100 microM vanadate, although the effects of Mn2+ and Co2+ (100 microM) were inhibited by vanadate. Also, the inhibition of the Mn2+ and Co2+ effects by vanadate was not seen in the presence of regucalcin. Moreover, regucalcin (0.5 microM) increased significantly the enzyme activity in the absence of Ca2+. This effect of regulcalcin was not altered by increasing concentrations of Ca2+ added, indicating that the regucalcin effect does not depend on Ca2+. The present results suggest that regucalcin activates directly (Ca(2+)-Mg2+)-ATPase in liver plasma membranes, and that the activation is not involved in the Ca(2+)-dependent phosphorylation of the enzyme. PMID- 8232288 TI - Characterization of the superinduction of the c-myc proto-oncogene in fibroblasts by benzamide derivatives. AB - In mouse fibroblasts stimulated from quiescence into proliferation by serum the induction of expression of the c-myc proto-oncogene is strongly stimulated by 3 methoxybenzamide. Similar superinduction effects are seen with related compounds such as 3-aminobenzamide and the acid analogues, 3-anisic acid and 3-aminobenzoic acid. Whereas the benzamide derivatives are inhibitors of poly(ADP-ribose) polymerase the acid analogues are not, suggesting that inhibition of this enzyme is not the basis for superinduction of the c-myc gene. Analysis of the kinetics of induction of c-myc mRNA indicates that the RNA accumulates more rapidly as well as to a higher level in the presence of serum plus 3-methoxybenzamide than with serum alone. However the stimulation is transient in both cases. Addition of actinomycin D at 30 min or 1 h after serum stimulation shows the c-myc mRNA to be stable at these times, in the presence or absence of 3-methoxybenzamide. Thus the effect of the latter on c-myc mRNA accumulation is likely to be exerted at the level of transcription or RNA processing rather than turnover of the mRNA. PMID- 8232289 TI - Signal transduction in Trypanosoma cruzi: opposite effects of adenylcyclase and phospholipase C systems in growth control. AB - Fetal calf serum (FCS), which is mitogenic for the pathogenic protozoa T. cruzi, inhibits cAMP production in basal and forskolin-stimulated epimastigotes. It also activates phosphoinositides hydrolysis yielding diacylglycerol and inositol phosphates (Ins-P). Ins-P production is enhanced by AlF4-, GTP or beta-gamma methylene-GTP, thus implying G proteins mediation in the phenomenon. An enzyme with phospholipase C activity which may be involved in the phospholipid metabolism was partially characterized. PMID- 8232290 TI - The biology of yeast mitochondrial introns. PMID- 8232291 TI - Definition of the Th/To ribonucleoprotein by RNase P and RNase MRP. AB - We show that the Th/To ribonucleoprotein is defined by (i) the co immunoprecipitation of two RNAs, (ii) the co-immunoprecipitation of four major polypeptides and (iii) the quantitative immune recognition of both RNase P and RNase MRP. No serum was found that recognizes either one of these two enzymes exclusively. The specific co-immunoprecipitation of RNase MRP and RNase P by all Th/To ribonucleoprotein autoantibodies indicates that the anti-Th/To autoimmune response is directed against both enzymes in a quantitatively indistinguishable manner. Thus the Th/To ribonucleoprotein is defined by RNase P and RNase MRP. PMID- 8232292 TI - Transcription of mononucleosomal particles acetylated in the presence of n butyrate. AB - Although a correlation between chemical acetylation of the amino-terminal tails of core histones and stimulation of RNA synthesis has been reported for nucleosomal core particles (Pineiro et al. (1991) Biochem. Biophys. Res. Commun. 177:370), no differences in transcription are detected between acetylated and nonacetylated mononucleosomal particles obtained from HeLa cells in the presence and absence of n-butyrate. Apparently, the lysine residues modified in the presence of n-butyrate are not the same responsible for the observed acetylation induced transcription. The acetylation obtained with n-butyrate might be significantly different from that present in transcriptionally active chromatin. PMID- 8232293 TI - Linear-dichroism spectroscopy for the study of structural properties of proteins. AB - This review gives an experiment directed survey of the application of linear dichroism (LD) spectroscopy to the study of proteins. LD spectroscopy is a relatively simple technique that provides information on the orientation of chromophores in molecules, on molecular characteristics such as shape, size and electronic properties, and on binding parameters in molecular complexes. Since LD is only observed when the molecules are non-randomly oriented in the sample, particular attention is paid to various orientation techniques, viz. in electric and flow fields, in polymer films and gels, and by light induction (photoselection). Examples are given on bacteriorhodopsin and retinals, chlorosomes, lens crystallins, aspartate aminotransferase, and the interaction of gene32- and recA-protein with DNA. PMID- 8232294 TI - Protein composition of mammalian spliceosomal snRNPs. PMID- 8232295 TI - Localisation of splicing snRNPs in mammalian cells. PMID- 8232296 TI - Structure and function of signal recognition particle (SRP). PMID- 8232297 TI - RNA-protein interactions in mRNA 3'-end formation. PMID- 8232298 TI - The hnRNP proteins. PMID- 8232299 TI - Nucleocytoplasmic transport and snRNP assembly. PMID- 8232300 TI - Mammalian protein factors involved in nuclear pre-mRNA splicing. PMID- 8232301 TI - Yeast protein splicing factors involved in nuclear pre-mRNA splicing. PMID- 8232302 TI - T-type Ca2+ channels are required for adrenocorticotropin-stimulated cortisol production by bovine adrenal zona fasciculata cells. AB - The function of low voltage-activated T-type Ca2+ channels in ACTH-stimulated cortisol production by bovine adrenal zona fasciculata cells (AZF) was explored in patch clamp and secretion studies. Nearly all AZF cells expressed only a low voltage-activated T-type Ca2+ current (IT) that was blocked by the diphenylbutylpiperidine (DPBP) Ca2+ antagonists penfluridol and pimozide with IC50S of 0.3 and 0.5 microM, respectively. Dihydropyridine (DHP) Ca2+ antagonists, including nimodipine, nisoldipine, and felodipine, also blocked T type Ca2+ current with IC50S ranging from 3.5-8.8 microM. Inhibition of IT by DPBP and DHP antagonists was voltage and use dependent. ACTH (1 nM) stimulated large (> 50-fold) increases in cortisol production by AZF cells, which were inhibited by Ca2+ antagonists at concentrations similar to those which blocked IT. Inhibition of cortisol production by Ca2+ antagonists was specific; ACTH induced insulin-like growth factor-I production by AZF cells was not affected by DPBP antagonists. The L channel-specific DHP Ca2+ agonist (-)Bay K 8644 did not enhance basal or ACTH-stimulated cortisol synthesis. These results demonstrate that functional T-rather than L-type Ca2+ channels are required for ACTH stimulated cortisol synthesis. They also suggest that these low voltage-activated channels, acting as the primary pathway for Ca2+ entry into AZF cells, couple ACTH-stimulated membrane depolarization to steroid hormone production. PMID- 8232303 TI - Glucose regulation of transforming growth factor-alpha expression is mediated by products of the hexosamine biosynthesis pathway. AB - We have recently shown that glucose and glucosamine regulate the transcription of transforming growth factor-alpha (TGF alpha) in rat aortic smooth muscle (RASM) cells. Based on the increased potency of glucosamine compared to glucose, we hypothesized that stimulation of TGF alpha transcription by glucose is mediated through the hexosamine biosynthesis pathway. The yeast cDNA for the rate-limiting enzyme of this pathway, glutamine:fructose-6-phosphate amidotransferase (GFA), was therefore expressed in RASM cells. GFA-transfected cells showed an increase in GFA activity, exhibiting a 2.2-fold increase in the synthesis of glucosamine-6 phosphate, the first product of the hexosamine biosynthetic pathway. To test the effect of GFA overexpression on TGF alpha transcriptional activity, cells were transiently cotransfected with GFA along with a reporter plasmid containing the firefly luciferase gene under control of the TGF alpha promoter. GFA-transfected cells exhibited a glucose-dependent 2-fold increase in TGF alpha activity compared to control cells. Maximal stimulation of TGF alpha-luciferase activity by glucosamine, however, was equivalent in GFA-and control-transfected cells, confirming that the stimulation observed by both agents operated through the same pathway. This increase in TGF alpha activity was inhibited (85% at 0.5 mM glucose and 69% at 30 mM glucose) by the glutamine analog and inhibitor of GFA, 6-diazo-5 oxonorleucine (10 microM). Control studies confirmed that the increased TGF alpha luciferase activity in the GFA-expressing cells was not an artifact of altered growth, survival, or transfection efficiency.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232304 TI - Human thyroxine-binding globulin gene: complete sequence and transcriptional regulation. AB - T4-binding globulin (TBG) is a glycoprotein of hepatic origin which transports thyroid hormone in serum. To characterize the human TBG (hTBG) gene, we studied its genomic organization, promoter activity, and regulation. To this purpose, we isolated from liver a complete hTBG cDNA clone containing the 5'-untranslated region and localized the transcription start site (TSS). The analysis of genomic clones revealed that the hTBG gene consists of five exons and that its exon intron organization is similar to that of other members of the serine protease inhibitor family. The first exon (exon 0) is a short noncoding sequence located 1.62 kilobase pairs (kbp) upstream from exon 1. Potential cis-acting transcriptional regulatory elements including a TATA box, a CAAT box, and a hepatocyte nuclear factor-1 binding motif were identified in the upstream region. A reporter gene in which 3.2 kbp of the 5'-flanking region, including exon 0, was inserted upstream of the bacterial chloramphenicol acetyltransferase gene showed significant activity when transfected into a hepatblastoma-derived (HepG2) cell line. The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate, down-regulated the promoter activity by more than 80% and completely inhibited hTBG synthesis, whereas thyroid hormone, glucocorticoid, estrogen, and nicotinic acid had little, if any, effect. A series of 5'-deletions revealed that the fragment -218 to +4 from the TSS had the highest promoter activity, nearly 1000-fold greater than the promoterless chloramphenicol acetyltransferase construct. When nonhepatocyte derived cell lines (CV-1 and CHO) were tested, promoter activity was reduced by a factor of 100, showing that the promoter works in liver-specific manner. The region -218 to -102 contains liver-specific enhancer elements, since deletion to nucleotide -101 resulted in a profound reduction of the promoter activity in HepG2 cells but not in CV-1 or CHO cells. On the other hand, mutational disruption of the putative hepatocyte nuclear factor-1 site (located 65 bp upstream of the TSS) completely abolished the promoter activity in all cell lines, indicating that this site is absolutely required for the transcription of the hTBG gene. PMID- 8232305 TI - Recombinant activation domains of virion protein 16 and human estrogen receptor generate transcriptional interference in vitro by distinct mechanisms. AB - Overexpression of transcriptional activators in transfection assays may inhibit their own activity or interfere with trans-activation by different sequence specific transcription factors. In this study we show that this phenomenon of transcriptional interference (squelching) can be mimicked in vitro by adding recombinant activation domains to nuclear extracts. We demonstrate that the acidic activation domain of virion protein 16 interferes both with basal transcription from TATA-box promoters and promoters activated by various trans activators in two different mammalian cell-free transcription systems. This suggests that virion protein 16 interacts with and thereby sequesters a basal transcription factor. In contrast the recombinant activation function 2 (AF-2) of human estrogen receptor does not affect basal promoter activity but inhibits TATA promoters activated by human progesterone receptor (hPR) or Sp 1 as well as the beta-globin and adenovirus major late promoter. By analyzing the effects of AF-2 on DNA binding of hPR and Sp1 we found that AF-2 inhibits the DNA binding activity of hPR, but not Sp1. Our data suggest that the recombinant AF-2 squelches Sp1 trans-activation by sequestering a common coactivator(s), whereas hPR function might be inhibited due to competition for a common cofactor stabilizing hPR dimers or through the formation of inactive heterodimers between AF-2 and hPR. PMID- 8232306 TI - Genomic elements involved in transcriptional regulation of the rabbit surfactant protein-A gene. AB - Expression of the surfactant protein-A (SP-A) gene is lung specific and is developmentally and hormonally regulated in fetal lung tissue. Cyclic AMP analogs and glucocorticoids stimulate transcriptional activity of the SP-A gene in fetal rabbit lung tissue in culture; an additive effect is observed when the agents are added in combination. To analyze the genomic regions that regulate SP-A promoter activity, fusion genes comprised of -1766, -991, -378, and -47 basepairs (bp) of DNA flanking the 5'-end of the SP-A gene, the transcription initiation site, and 20 bp of exon I linked to the human GH (hGH) structural gene were subcloned into a replication-defective human adenovirus vector and transfected into differentiated rat type II cells in primary culture. SP-A promoter activity was analyzed by RIA of hGH protein in the culture medium. In type II cells transfected with SP-A-1766:hGH and SP-A-991:hGH fusion genes, hGH production was induced 30- to 40-fold by (Bu)2AMP (Bt2cAMP; 1 mM). When type II cells were transfected with the SP-A-378:hGH fusion gene, basal levels of expression were reduced by more than 50%; however, Bt2cAMP caused an 11-fold increase in hGH production. In type II cells transfected with the SP-A-47:hGH fusion gene, basal levels of hGH production were essentially undetectable, and no stimulatory effect of Bt2cAMP was apparent. Cyclic AMP stimulation of expression of the SP-A 1766:hGH, SP-A-991:hGH, and SP-A-378:hGH fusion genes was limited to type II pneumonocytes in primary culture and was absent in two lung adenocarcinoma cell lines (NCl-H358 and A549), which do not express SP-A, and in cAMP-responsive adrenal Y1 cells. Mutations of a putative cAMP-responsive element (TGACCTCA) at 261 bp revealed its functional importance in mediating cAMP regulation of SP-A gene expression. Unexpectedly, dexamethasone (Dex; 10(-7) M) antagonized the stimulatory effect of Bt2cAMP on expression of SP-A:hGH fusion genes containing from -378 to -1766 bp of 5'-flanking DNA as well as that of a fusion gene construct containing -991 bp of 5'-flanking DNA, the first exon, the first intron, and 20 bp of the second exon (SP-A-991+670:hGH). The inhibitory effect of Dex was dose dependent, with half-maximal inhibition occurring at a Dex concentration of 8 x 10(-10) M. The inhibitory effect of Dex was prevented by the glucocorticoid receptor antagonist RU486.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8232307 TI - Citation for the Fred Conrad Koch Award of The Endocrine Society to Jean D. Wilson. PMID- 8232308 TI - Citation for the Edwin B. Astwood Lecture Award of The Endocrine Society to Ronald M. Evans. PMID- 8232309 TI - Citation for the Rhone-Poulenc Rorer Clinical Investigator Lecture Award of The Endocrine Society to William L. McGuire. PMID- 8232310 TI - Citation for the Sidney H. Ingbar Distinguished Service Award of The Endocrine Society to Gabriel Bialy. PMID- 8232311 TI - Citation for the Gerald D. Aurbach Award of The Endocrine Society to R. Daniel Camerini-Otero. PMID- 8232312 TI - Citation for the Robert H. Williams Distinguished Leadership Award of The Endocrine Society to Willard P. VanderLaan. PMID- 8232313 TI - Citation for the Ernst Oppenheimer Award of The Endocrine Society to Margaret A. Shupnik. PMID- 8232314 TI - Citation for the Richard E. Weitzman Memorial Award to Daniel J. Drucker. PMID- 8232315 TI - Functional domains and phosphorylation of the orphan receptor Nur77. AB - Nur77 represents a unique class within the steroid receptor superfamily since its synthesis is tightly regulated by extracellular signals and it is capable of potent transactivation activity in the absence of an exogenously added ligand. In this study, we sought to dissect the functional domains regulating the activities of Nur77 by deletion mapping. We demonstrate that whereas the transactivation activity of Nur77 resides in the amino-terminal domain, the carboxy-terminal domain regulates this activity. A short deletion from the carboxy terminus eliminates transactivation activity while a further deletion restores the activity. Deletion of the domain immediately carboxyl to the zinc fingers motif eliminates both DNA binding activity and nuclear localization, thus abolishing transactivation. Nur77 is posttranslationally modified predominantly by phosphorylation, which occurs primarily at the N-terminal domain. The growth related kinase pp90rsk, but neither the pp44mapk nor the pp70s6k, can phosphorylate recombinant Nur77 in vitro. Furthermore, we have identified a site within the region required for sequence-specific DNA binding, Ser-354, that is phosphorylated by pp90rsk in vitro; this site is also phosphorylated in vivo. The possibility that phosphorylation might affect DNA binding is discussed. PMID- 8232316 TI - Okadaic acid, a protein phosphatase inhibitor, enhances transcription of a receptor gene containing sequence A of the human prolactin promoter. AB - Human PRL (hPRL) gene expression is controlled by cAMP and Ca2+. This control is mediated by two cis-elements: a Pit-1 binding site (-62 to -35) and sequence A ( 110 to -85), present in the hPRL promoter. We have investigated whether protein phosphatases could be involved in this regulation. GC-type rat pituitary tumor cells were transfected with sequence -138 to -35 of the hPRL gene promoter, upstream from a thymidine kinase promoter and a chloramphenicol acetyltransferase (CAT) reporter gene. Addition of okadaic acid (OA), a specific inhibitor of protein phosphatases 1 and 2A, stimulates transient expression of the CAT gene. The dose-response curve shows a maximal effect at 25 nM OA (2.2-fold stimulation above controls). The OA effect is also observed with a natural 4500-base pair hPRL promoter. A single copy of the hPRL promoter sequence -115 to -85 (sequence A) confers to a thymidine kinase-CAT construct an identical response to OA, whereas a single copy of the proximal Pit-1 binding site does not. Synergism is observed between cAMP and OA in activating PRL gene transcription. This synergism is also observed with a single copy of sequence A. The effect of cAMP is not mediated by an L-type Ca2+ channel, since addition of the Ca2+ channel antagonist verapamil does not decrease it, nor does complexing extracellular Ca2+ significantly reduce it. Furthermore, OA and the Ca2+ channel opener BAY K8644 exert additive effects. PMID- 8232317 TI - A novel in vitro model for studying signal transduction and gene regulation via the growth hormone receptor. AB - Buffalo rat liver cells were stably transfected with an expression vector containing rat GH (rGH) receptor cDNA. Transfected cells expressed rGH receptor mRNA and specifically bound GH with high affinity. When transfected cells were stimulated with GH, levels of lipoprotein lipase (LPL) mRNA were increased in a time- and dose-dependent fashion, while glyceraldehyde-3-phosphate-dehydrogenase mRNA levels were unaffected. No GH binding or LPL mRNA could be detected in untransfected cells. Treatment of transfected cells with actinomycin D inhibited the GH-stimulated increase in LPL mRNA, indicating that GH acts at a transcriptional level. When protein synthesis was inhibited using cycloheximide, basal levels of LPL mRNA were increased, and there was no GH stimulation. This suggests that LPL gene expression is constantly repressed by a labile protein. Chloramphenicol acetyltransferase constructs containing the human LPL promoter could be regulated by GH. In conclusion, stimulation of the rGH receptor in stably transfected Buffalo rat liver cells results in specific induction of LPL gene expression. This provides a novel model to study the mechanism of GH action, particularly in relation to gene regulation. PMID- 8232318 TI - The rat proenkephalin germ line promoter contains multiple binding sites for spermatogenic cell nuclear proteins. AB - Rat and mouse spermatogenic cells contain a family of 1700-nucleotide (nt) proenkephalin mRNAs that are generated from an alternate, germ cell-specific promoter. This promoter is located approximately 350 base pairs (bp) downstream of the promoter used in somatic cells, within the first intron for the somatic transcript. In a previous study, rat proenkephalin-chloramphenicol acetyltransferase fusion genes containing both promoters were shown to be transcribed selectively from the germ cell promoter and in the correct developmental pattern in spermatogenic cells of transgenic mice. In the present study it was found that spermatogenic cell-specific transgene expression was maintained after deletion of the upstream somatic promoter. This result establishes that the rat proenkephalin germ-line promoter is capable of functioning independently of transcriptional elements associated with the somatic promoter and localizes the requisite spermatogenic cell cis-elements to a 500-bp region encompassing the germ cell initiation sequences. A comprehensive analysis of binding sites for rat spermatogenic cell nuclear factors within this 500-bp region was performed using gel-shift and DNAse I footprinting techniques. Eight distinct binding regions were identified, each of which formed one or more cell specific complexes with nuclear proteins from rat spermatogenic cells. These results suggest that multiple cis-acting elements may cooperate in the cell specific and developmental regulation of rat proenkephalin gene transcription during spermatogenesis. PMID- 8232319 TI - Peptide growth factors elicit estrogen receptor-dependent transcriptional activation of an estrogen-responsive element. AB - Epidermal growth factor (EGF) elicits estrogen receptor (ER)-dependent physiological sequelae and estrogen-like biochemical effects on the ER in the mouse uterus. These in vivo observations indicate that EGF may elicit some of its actions by activation of the ER. The effect of peptide growth factors on activation of a consensus estrogen-responsive element was assessed in a strain of Ishikawa human endometrial adenocarcinoma cells with negligible levels of ERs, as determined by Western blot and [3H]estradiol binding, and in BG-1 human ovarian adenocarcinoma cells, which contain abundant ERs. EGF and transforming growth factor-alpha induced transcriptional activation of a consensus ERE in an ER dependent manner in both cell types. Transcriptional activation by the growth factors was inhibited by ICI 164,384, an ER receptor antagonist, and neutralizing antibodies to the EGF receptor. Immunodetection of the ER in BG-1 cells demonstrated that receptor levels were not induced by transforming growth factor alpha vs. untreated cells. ER deletion mutants containing amino acids 1-339 and 121-599 were transfected into Ishikawa cells. The 1-339 mutant was more active in inducing transcription after EGF treatment than the 121-599 mutant. Estrogen only stimulated transcription in the presence of the 121-599 mutant, while 1-339 was inactive. Interestingly, synergism between a physiological dose of estrogen and peptide growth factors was observed. The presence of cross-talk between EGF receptor and ER signaling pathways suggests that interactions between growth factors and steroid receptors may modulate hormonal activity influencing normal and aberrant function in mammalian cells. PMID- 8232320 TI - Down-regulation of calcitonin gene transcription by vitamin D requires two widely separated enhancer sequences. AB - Transcription of the calcitonin (CT) gene is down-regulated by vitamin D in normal and transformed thyroid C cells. DNA transfer techniques have been previously used to map and characterize a cAMP-induced enhancer at nucleotides 255 to -129 and an enhancer of basal transcription at -1060 to -905 in the CT 5' flanking DNA. The same methods were used to identify a negative response element for vitamin D. Deletion mutants of a genomic fragment of CT extending from nucleotides -1460 to +90 were attached to a promoterless GH gene and transfected individually into the medullary thyroid carcinoma cell line TT. CT nucleotides 1460 to -129 induced significant basal transcription of the GH reporter gene in TT cells. Basal transcription was elevated 3-fold to 4-fold by treatment with cAMP analog. The biologically active metabolite of vitamin D3, 1,25 dihydroxyvitamin D3, had a minor (20%) inhibitory effect on basal transcription but inhibited more than 60% of the cAMP-induced transcription. We further investigated the cAMP-induced response and found that transcriptional activity of the downstream cAMP-induced enhancer was greatly synergized in the presence of the upstream enhancer of basal transcription. The latter enhancer contained three functional CANNTG sequences designated E1 (nucleotides -1060 to -1030), E2 (nucleotides -940 to -920), and E3 (nucleotides -920 to -900). E2 and E3 were essential for maximal cAMP-induced transcription. Detailed mapping of the vitamin D response showed that a minimum requirement for inhibition of the cAMP-induced enhancer by vitamin D was a sequence overlapping E3 (nucleotides -920 to -829). We conclude that a negative response element to vitamin D is located between nucleotides -920 and -829 in the CT 5' flanking DNA. It is possible that vitamin D inhibits transcription by interfering with the synergistic interaction between the cAMP-induced enhancer and the enhancer of basal transcription. PMID- 8232321 TI - Mutational analysis of active site contact residues in anti-fluorescein monoclonal antibody 4-4-20. AB - The contribution to high affinity Fl binding by each crystallographically defined Mab 4-4-20 (Ka = 1.7 x 10(10) M-1; Qmax = 90%) ligand contact residue (L27dHis, L32Tyr, L34Arg, L91Ser, L96Trp and H33Trp) has been determined by site-specific mutagenesis studies. All six antigen contact residues were changed to Ala in the single-chain derivative of Mab 4-4-20 and following expression in E. coli, denaturation, refolding and purification, each SCA mutant was characterized in terms of Fl binding affinity, Qmax, lambda max and idiotype. Results demonstrated that Ala substitutions at each ligand contact residue reduced the binding affinities and quenching maxima for all residues except L27d which retained wild type characteristics. The SCA TyrL32Ala, SerL91Ala and TrpH33Ala mutants exhibited binding affinities that were approximately 1000-fold lower than the wild type value and greatly reduced Qmax values. Additionally, other amino acid substitutions were performed at three of the six antigen contact residues (L91Ser, L96Trp and H33Trp) to further evaluate the role of each in Fl binding. Therefore, the following mutations were constructed and characterized: SerL91Asn, TrpL96Tyr, TrpL96Phe, TrpL96Leu, TrpH33Tyr and TrpH33Phe. Results of site specific mutagenesis studies are discussed in terms of Mab active site structure and suggest that L32Tyr, L91Ser and H33Trp are important for high affinity Fl binding and efficient Fl quenching. PMID- 8232322 TI - Genetically engineered deglycosylation of the variable domain increases the affinity of an anti-CD33 monoclonal antibody. AB - M195 is a murine monoclonal antibody that binds to the CD33 antigen and is being tested for the treatment of myeloid leukemia. Surprisingly, a complementarity determining region (CDR)-grafted, humanized M195 antibody displayed a several fold higher binding affinity for the CD33 antigen than the original murine antibody. Here we show that the increase in binding affinity resulted from eliminating an N-linked glycosylation site at residue 73 in the heavy chain variable region in the course of humanization. Re-introducing the glycosylation site in the humanized antibody reduces its binding affinity to that of the murine antibody, while removing the glycosylation site from the murine M195 variable domain increases its affinity. The removal of variable region carbohydrates may provide a method for increasing the affinity of certain monoclonal antibodies with diagnostic and therapeutic potential. PMID- 8232323 TI - Structure of complement poly-C9 determined in projection by cryo-electron microscopy and single particle analysis. AB - The ring-like complement 'lesions' found on membranes of complement lysed cells comprise a complex of components C5b through C9 that coalesce to form hollow cylinders which penetrate the membrane bilayer and create lytic pores. Walls of these C5b-9 membrane attack complex cylinders may consist primarily of the C9 component, since samples of purified, isolated C9 can polymerize into cylindrical structures which appear identical with the fully assembled C5b-9 complex. The structure of these poly-C9 molecules has been investigated using the techniques of cryo-electron microscopy and single particle analysis. Sets of single poly-C9 particles viewed as rings were selected from cryo-EM images, then particles were aligned and treated by correspondence analysis to identify the principle interparticle similarities and variations. The highest ranking variation found was the presence or absence of a dense inner ring of protein density. Other important variations were interpreted as different types of particle tilt. These results were used in selecting a subgroup of untilted particles for averaging and symmetry analysis. The rotational power spectrum of the initial average suggested 13-fold symmetry. The 13-fold symmetry was used to select and group particles for further analysis. Individual particles were 13-fold rotational averaged and those with enhanced peripheral features were placed into either a right-handed subgroup or into a left-handed subgroup based on orientation of the peripheral features. Particles within each group were aligned and averaged, and a poly-C9 structure was produced which shows important structural details and from which the C9 monomer structure can be deduced. The poly-C9 structure contains a dense inner ring of diameter between 113-181 A and which is modulated into 13 discrete peaks with peak-to-peak separation of approx. 35 A. The dense inner ring is surrounded by a less dense, concentric outer rim extending to 254 A diameter. The outer rim contains projections that are contiguous with the inner peaks but are skewed relative to the ring radius to produce the appearance of a pin-wheel. These projections correspond with the peripheral features picked up in the rotationally averaged individual particles; the left- or right-handed orientation of projections may result from the up/down orientation of individual particles in ice. The C9 monomer structure within the cylinder is suggested by the density distribution. The monomer would be a rod with diameter of 35 A, oriented parallel to the cylinder axis and would be roughly perpendicular to a membrane.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8232324 TI - N sequences, P nucleotides and short sequence homologies at junctional sites in VH to VHDJH and VHDJH to JH joining. AB - Junctional sequences in VH to VHDJH and VHDJH to JH joining occurring in Abelson virus-transformed immature B cell lines were PCR-amplified and sequenced. In VH to VHDJH joining, 24 (23%) out of 105 junctions examined here had P nucleotides and/or N sequences, and out of the remaining 81 junctions without P nucleotides and N sequences, 57 (70%) had short sequence homologies of one or two bases (A, C, G, CA or AG) and three had long sequence homologies at the junctional sites. In VHDJH to JH joining, 38 (43%) out of 89 junctions examined here had P nucleotides and/or N sequences, and out of the remaining 51 junctions without P nucleotides and N sequences, 47 (92%) had short sequence homologies of one or two bases (C, T, G, TG or GG) at the junctional sites. These results indicate that short sequence homologies play an important role for end joining in VH to VHDJH and VHDJH to JH joining. PMID- 8232325 TI - The 3' half of the mouse mammary tumor virus orf gene is not sufficient for its superantigen function in transgenic mice. AB - The Mouse Mammary Tumor Virus (MMTV) long terminal repeat contains an open reading frame (orf) of 960 nucleotides encoding a 36 kDa polypeptide with a putative transmembrane domain and five N-glycosylation sites in the N-terminal part of the protein. Transgenic mice bearing either the complete or the 3' terminal half of the orf sequence of MMTV-GR under the control of the SV40 promoter were raised. As shown previously by FACS analysis transgenic mice which express the complete orf gene have a significant deletion of V beta 14 expressing T cells at 6 weeks of age. Here we show that no clonal deletion of V beta 14 bearing T cells takes place in transgenic mice that contain orf sequences from the fifth ATG to the termination codon. The pattern of tissues expressing the truncated transgene was studied by the Polymerase Chain Reaction (PCR) and was very similar to the one obtained in the V beta 14 deleting animals. These data suggest that the amino-terminal portion of the ORF protein (pORF) is required for a superantigen function, while our previous data indicated that determinants from the carboxy-terminus play an important role for TCR V beta specificity. PMID- 8232326 TI - Expression of a mouse-channel catfish chimeric IgM molecule in a mouse myeloma cell. AB - Fusion genes encoding a murine VH domain and the constant region domains of the mu chain from the channel catfish, Ictalurus punctatus, were stably expressed in the lambda light chain producing mouse myeloma cell line J558L. Although the pathways of pre-mRNA processing for expression of membrane (micron and secreted (microsecond) forms of the mu chain differ between mammals and teleosts, mRNAs encoding both catfish micron and microsecond were correctly expressed in the mouse myeloma cells. The mouse-channel catfish chimeric mu chain polypeptide was able to associate covalently with the mouse lambda light chain and assemble, intracellularly, into polymers of covalent structure (microL)2-8 which resembled those seen with native catfish IgM. In contrast to native catfish IgM, the mouse catfish chimeric IgM showed the property of binding strongly to protein A of Staphylococcus aureus. The mouse-channel catfish chimeric IgM was core glycosylated, but did not contain terminal sialic acid. Secretion rates for the chimeric IgM were low, and the possibility could not be excluded that extracellular chimeric IgM was released from dead or dying cells. The reason(s) for the intracellular retention of the chimeric IgM (probably in the endoplasmic reticulum) are not known, but those mechanisms involving retention via cysteine residues were excluded. PMID- 8232327 TI - Human IgG3 can adopt the disulfide bond pattern characteristic for IgG1 without resembling it in complement mediated cell lysis. AB - In this paper we describe the construction of mouse-human IgG3 mutant antibodies resembling IgG1 in their disulfide bond pattern between the heavy and light chain (H-L) and between the two heavy chains (H-H). The effector functions of these mutant antibodies were compared to normal IgG3 and IgG1. Changing only the disulfide bond pattern between the heavy and light chains did not alter the ability to induce complement mediated cell lysis (CML), regardless of the amount of corresponding antigen that had been introduced to the surface of the target cells. However, alteration of the disulfide bond pattern between the two heavy chains had a large effect on CML due to shortening of the hinge from 62 to 15 amino acids. No difference between the mutants and normal antibodies in antibody dependent cell-mediated cytotoxicity (ADCC) was observed. This suggests that IgG3 can adopt the H-L disulfide bond pattern of IgG1 without obtaining the CML activity characteristic for IgG1. PMID- 8232328 TI - Membrane immunoglobulin without sheath or anchor. AB - The canonical form of the B cell antigen receptor is composed of membrane immunoglobulin sheathed by the alpha/beta heterodimer. Whereas membrane IgM cannot be transported to the cell surface in the absence of alpha/beta, both IgD and IgG2b can be expressed naked (i.e. without alpha/beta) on the surface of myeloma transfectants. In the case of one cell-line, such naked IgD has been shown to be inserted into the membrane by a glycosyl-phosphatidylinositol anchor. Here, however, we show that both IgD and IgG2b (but not IgM) can be expressed on the surface of myeloma transfectants without either sheath or anchor. This distinction between the isotypes is attributable to differences in the region of the transmembrane segment. PMID- 8232329 TI - Isolation and characterization of a new murine MHC class II transcription mutant cell line. AB - We have isolated and characterized a new MHC class II transcription mutant cell line, called UV. This cell line was derived from the mouse B lymphoma A20 by UV light-induced mutagenesis and immunoselection for the loss of surface MHC class II molecules. It expresses only 5% of the level of MHC class II molecules on A20 and this is associated with a similar reduction of class II specific mRNA. This defect cannot be restored by the MHC class II transcription inducers, IL-4 and IFN gamma, confirming that the mutation acts at the transcription level. The mutation also affects MHC class I expression, but the transcription of class I molecules is not affected. In contrast, the expression of other markers, such as the invariant chain and the surface immunoglobulins G and M, is not modified. Such a variant should prove useful for the study of the transcription factors involved in the regulation of MHC class II expression. PMID- 8232330 TI - Construction and initial characterization of a mouse-human chimeric anti-TNF antibody. AB - Tumor necrosis factor-alpha (TNF) has been implicated in the pathogenesis of a variety of human diseases including septic shock, cachexia, graft-versus-host disease and several autoimmune diseases. Monoclonal antibodies directed against TNF provide an attractive mode of therapeutic intervention in these diseases. We have generated a murine monoclonal antibody (A2) with high affinity and specificity for recombinant and natural human TNF. To increase its therapeutic usefulness, we used genetic engineering techniques to replace the murine constant regions with human counterparts while retaining the murine antigen binding regions. The resulting mouse-human chimeric antibody should have reduced immunogenicity and improved pharmacokinetics in humans. Molecular analysis of light chain genomic clones derived from the murine hybridoma suggests that two different alleles of the same variable region gene have rearranged independently and coexist in the same hybridoma cell. The chimeric A2 antibody (cA2) exhibits better binding and neutralizing characteristics than the murine A2 which was shown to contain a mixture of two kappa light chains. The properties of cA2 suggest that it will have advantages over existing murine anti-TNF antibodies for clinical use. PMID- 8232331 TI - Re-evaluation of the concept of functional affinity as applied to bivalent antibody binding to cell surface antigens. AB - Although it is recognized that the bivalent binding of an antibody to a multivalent antigen cannot be characterized by true affinity, "functional affinity" is frequently determined. "Functional affinity" is calculated by the same methods used for true affinity, and is presumed to have similar biological significance. The data presented herein demonstrate, for IgG antibodies binding to cell surface antigens, that "functional affinity" does not adequately describe the interactions that occur. First, the dissociation rate was not first-order, but rather reflected heterogeneity of bound Ab. A minority of bound Ab (probably monovalently bound) dissociated relatively rapidly, while the majority dissociated very slowly, and, for most purposes, should be considered irreversibly bound. Although irreversible binding is incompatible with a state of equilibrium, the dissociation rate was greatly increased in the presence of free Ab, which may explain why equilibrium is reached in a Scatchard-type experiment. In addition, "functional affinity" was found to vary depending on trivial experimental conditions, such as the volume of incubation. Thus, antibodies do not possess a single "functional affinity", but rather a spectrum of "functional affinities", which are therefore of little value in comparing different antibodies and cannot be used to predict the extent of binding under other experimental conditions. Both of these discrepancies can be attributed to the effect of bivalent Ab binding. We conclude that "functional affinity" determination is inappropriate. PMID- 8232332 TI - Variable region sequence and characterization of monoclonal antibodies to a N,N',N"-trisubstituted guanidine high potency sweetener. AB - A library of monoclonal antibodies (mAb) was made against a trisubstituted guanidinium sweetener (N-(p-cyanophenyl)-N'-(diphenylmethyl)guanidine acetic acid) that is 200,000 times sweeter than sucrose on a molar basis. The mAb were characterized in terms of their ligand affinities, H- and L-chain isotypes and V region amino acid sequences. Nine of these mAb were found to have dissociation constants in the nanomolar range. The H-chain V-regions were cloned, sequenced and found to be derived from five different families (Q52, X24, J558, 7183 and 36 60). L-chain V-regions were found to be derived from three kappa families (V kappa-4/5, V kappa-19/28 and V kappa-1) and one lambda family (V lambda-1). Amino acid homologies with these family sequences ranged from 51-91% for heavy chains and 69-97% for light chains. Sequence comparisons with Ig structures solved by X ray diffraction were made in order to identify canonical structures. Identification and localization of combining region tryptophans (L:96W and H:33W) for two mAb (NC10.8 and NC6.8) supported previous ligand-induced tryptophan fluorescence quenching observations. PMID- 8232333 TI - Role of homology-directed recombination: predominantly productive rearrangements of Vh81X in newborns but not in adults. AB - In the neonate, Ig V-D-J junctions often occur at regions of short sequence homology, resulting in one to two predominant junctional sequences for most V-D and D-J recombinations. We have proposed that this mechanism of homology-directed recombination may play a role in the non-random usage of VH genes observed in fetal and neonatal life, since use of the short homologies at V-D junctions would preferentially make productive rearrangements for the overutilized 7183 and Q52 VH genes, and would make predominantly non-productive rearrangements for the underutilized VHJ558 gene family. Here we test this hypothesis for the 81X gene from the VH7183 family. Since pre-B cells which have rearranged the 81X gene do not appear to undergo the normal clonal proliferation before light chain rearrangement, analysis of the percentage of productive versus non-productive rearrangements for this VH gene is not skewed by the expansion of pre-B cells with productively rearranged IgH alleles. If V-D-J rearrangements were random, one would predict that only one-third of the rearrangements would be in-frame. This is close to what we observed for the 81X gene in adult bone marrow. In contrast, we show that 62% of all 81X rearrangements in fetal/newborn pre-B cells were productive. Forty-one percent of all the neonatal pre-B sequences containing DFL16 or DSP2 used homology-directed recombination to create the predominantly observed V-D junctional sequences, and 93% of those sequences were productive. This is consistent with our hypothesis that the mechanism of homology-directed recombination would result in an increased proportion of productive 81X rearrangements in the newborn. Therefore, we suggest that in fetal and neonatal life, when N regions are lacking, VH7183 and VHQ52 genes are more likely to undergo productive rearrangements than other VH families and thus are much more likely to contribute to the early B cell repertoire. PMID- 8232334 TI - Immunoglobulin variable region sequences of two human monoclonal antibodies directed to an onco-developmental carbohydrate antigen, lactotetraosylceramide (LcOse4Cer). AB - A human monoclonal antibody, 11-50, was generated and was shown to recognize an onco-developmental carbohydrate antigen, LcOse4Cer. The isotype of this antibody was IgM, lambda, similar to the previously known human anti-LcOse4 antibodies, such as IgMWOO and HMST-1. We raised a murine anti-idiotypic antibody G3 (IgG1, kappa) against 11-50, and tested its reactivity towards the affinity purified human polyclonal anti-LcOse4 antibodies prepared from pooled human sera using a Gal beta 1-->3GlcNAc beta-immobilized column. The results indicated that at least a part of the human polyclonal anti-LcOse4 antibodies shared the G3 idiotype with 11-50. We further analyzed the sequence of variable regions of the two anti LcOse4 antibodies, 11-50 and HMST-1. Sequence analysis of the heavy chain variable regions indicated that the VH regions of these two antibodies were highly homologous to each other (93.5% at the nucleic acid level), and these antibodies utilized the germline genes VH1.9III and hv3005f3 as the VH segments, which are closely related germline genes of the VHIII family. It was noted that these germline VH genes are frequently utilized in fetal B cells. The JH region of both antibodies was encoded by the JH4 gene. For the light chain, the V lambda segments of the two antibodies were 96.3% homologous to each other at the nucleic acid level. The V lambda segments of both antibodies showed the highest homology to the rearranged V lambda gene called V lambda II.DS among reported V lambda genes, while the exact germline V lambda genes encoding the two antibodies were not yet registered in available sequence databanks. The amino acid sequences of the J lambda segments of both antibodies were identical. These results indicate that the two human antibodies recognizing the onco-developmental carbohydrate antigen Lc4 are encoded by the same or very homologous germline genes. PMID- 8232335 TI - Antibody recognition of the recombinant human nuclear antigens RNP 70 kD, SS-A, SS-B, Sm-B, and Sm-D by autoimmune sera. AB - Five human nuclear antigens, RNP 70 kD, SS-A, SS-B, Sm-B and Sm-D, were produced in E. coli using the expression vector pSEM. cDNAs encoding these antigens were ligated to a truncated lacZ' gene of the vector and the beta-galactosidase fusion proteins were efficiently expressed as intracellular inclusion bodies after isopropyl-beta-thiogalactopyranoside induction. The antibody reactivities of these fusion proteins were evaluated by Western blot and by ELISA employing panel sera from patients with autoimmune diseases such as systemic lupus erythematosus, Sjogren's syndrome or mixed connective tissue disease. The three fusion proteins, RNP 70 kD, SS-B, and Sm-B, showed good reactivities in both systems, whereas the other two fusion proteins, SS-A and Sm-D, showed poor and no reactivity in both systems, respectively. It can be concluded that RNP 70 kD, SS-B and Sm-B recombinant antigens are useful reagents for the differential diagnosis of the autoimmune diseases. PMID- 8232336 TI - IgG auto- and polyreactivities of normal human sera. AB - Using a panel of self antigens, IgM autoreactivities were clearly and constantly detected by enzyme immunoassay (EIA) in the sera of 29 normal human individuals. Similarly, IgM autoreactivities in sera were reproducibly detected by immunoblotting, using human organ extracts as the antigen sources. In contrast, IgG reactivities were low in whole sera but were considerably increased after affinity-chromatography purification on protein G-Sepharose. These increases differed from one individual IgG preparation to another and from one antigen to another (from 1-94 times) resulting in a unique IgG autoreactivity pattern for each subject. IgG reactivities diminished markedly when the IgG-depleted serum was added to the isolated autologous IgG. IgM antibodies isolated from sera on F(ab')2 IgG immunoadsorbent partially inhibited the binding of IgG to tubulin and myosin but not to actin. The individual IgG preparations examined separately exhibited, with all the autoantigens of the panel, higher autoreactivities than those of the same-but-pooled IgGs, which in turn were higher than those of a commercially available human IgG preparation obtained from approximately 8,000 healthy donors and used for intravenous injection. Depending upon the individual IgG sample, 31-65% of the IgG were bound to a DNP-Sepharose column and were eluted with DNP-glycine. The isolated anti-DNP antibodies were found to be polyreactive and possess higher autoreactivities than the original IgG preparation for all the antigens of the panel. Similarly, IgG antibodies analysed using an antibody exchange procedure were found to be essentially polyreactive but some apparently monospecific antibodies were also noted. These results suggest that the great majority of IgG present in normal humans are composed of polyreactive autoantibodies. IgG autoreactivities are only marginally expressed in these whole sera because of IgM-IgG, IgG-IgG and other, still unidentified, interactions. PMID- 8232337 TI - Bacterially expressed Fabs of monoclonal antibodies neutralizing tumour necrosis factor alpha in vitro retain full binding and biological activity. AB - Antibody fragments specific for the human tumour necrosis factor alpha (TNF alpha) have been cloned from lambda combinatorial expression libraries using total RNA obtained from three different hybridoma cell lines of therapeutic interest. The previously described bacteriophage lambda vectors, lambda HC2 and lambda LC1, were modified to create unique antibody cloning sites in the combinatorial construct and a novel tag peptide was inserted at the C-terminal end of the expressed Fd chain. Sequence analysis of the cloned Fabs indicated that two of them were derived from a single B cell. Expression in E. coli showed that the amount of recovered Fab in the bacterial culture medium was related to the sequences of the variable coding regions. Hybrid Fabs created by chain exchange of similar antibodies were as active as the originally paired Fabs in binding assays. The relative affinities and the capacities of the bacterially expressed Fabs to neutralize TNF alpha cytotoxicity in vitro were identical to those of the parental antibodies. The results demonstrate that, using an in vitro approach, it is possible to generate from existing hybridoma cell lines high affinity Fabs which retain antigen specificity. The cloning sites incorporated into the C-terminal parts of these Fabs will now permit their further modification to include additional functional characteristics not possible with the original hybridoma antibodies. PMID- 8232338 TI - Native and recombinant Fel dI as probes into the relationship of allergen structure to human IgE immunoreactivity. AB - To delineate the relationship between the structural conformation and the stability of an allergen and its antigenicity, we have chosen the major allergen from cat dander, Fel dI. From protein sequence analysis data we have examined the structure of the naturally occurring Fel dI and we have found it to exist as an anti-parallel heterodimer. We have used ELISA, RAST, Western blot and histamine release techniques to compare the IgE reactivity of a set of cat allergic patient samples to purified, native Fel dI and the E. coli expressed chains 1 and 2. Results from these studies demonstrate a significant level of IgE reactivity to all forms when examined for direct binding. However, both blot and ELISA competition assays show a much higher reactivity to Fel dI in solution compared to the separate recombinant chains and this is supported by the histamine release data. Although native Fel dI chain 2 contains an N-linked carbohydrate moiety, this does not seem to play a role in the reactivity of IgE to chain 2. Denaturation of Fel dI with alkali conditions leads to a dramatic decrease in IgE reactivity, even though measurable changes to the backbone structure of the protein are minimal. One proposed explanation is that both chains possess a core region determined by their primary structures and that the major IgE epitopes are dependent upon them. The relative reactivity amongst these allergen forms varied with the method of analysis, implying that the conformational requirements for IgE antibody binding are best studied by the application of more than one experimental protocol. Results from these qualitative analyses afford insight into the allergenicity of this exceptionally stable cat pelt protein. PMID- 8232339 TI - Variable region genes of anti-HIV human monoclonal antibodies: non-restricted use of the V gene repertoire and extensive somatic mutation. AB - The extent of the expressed human V gene repertoire for the most part has been derived from fetal cDNA libraries, autoantibodies, and myeloma proteins. In order to continue to explore the utilization of the VH and VL gene repertoire in response to exogenous viral antigens, the heavy and light chain cDNAs from four human anti-HIV monoclonal antibodies were PCR amplified from human-mouse heterohybridomas, cloned, and nucleotide sequence analysis performed. Of the monoclonals analyzed, three were directed against gp120 and one reacted with gp41. Three of the antibodies were of the IgG1 lambda isotype and one was an IgG1 kappa. Three of the four heavy chains were derived from VHI gene segments and one VHII was observed. D segments showed evidence of D-D joining and three JH4 and one JH5 gene were utilized. Two V lambda II lambda chains and one from the V lambda III gene family were observed and the single kappa chain sequenced was from the V kappa III family. DNA sequence comparison with known germline gene segments identified putative precursor V gene segments for one of the heavy chains and two light chains. Comparison of the expressed amino acid sequences with the predicted germline sequences indicated that changes were clustered in the CDRs and FR3 regions of the V gene segments. We reported previously the nucleotide sequences of five human monoclonal antibodies from HIV-infected individuals, three of which utilized VHIV, one VHV and one a VHI gene segment and also found extensive evidence of somatic mutation. Collectively, our results indicate that an antigen driven response is functioning following HIV infection and, surprisingly, to date we have not encountered a VHIII gene segment. Since VHIII is the largest human VH gene family, it may well be that this under representation has both functional and clinical implications. PMID- 8232340 TI - Effects of IL-4 and Fc gamma receptor II engagement on Egr-1 expression during stimulation of B lymphocytes by membrane immunoglobulin crosslinking. AB - Egr-1 is an immediate early gene that is rapidly upregulated in response to mitogenic signals induced by antigen receptor crosslinking on murine B lymphocytes. It has been shown that levels of Egr-1 expression are closely correlated with B cell proliferation in several models of B cell activation and tolerance. We compared the expression of Egr-1 during B cell stimulation with Fab'2 and IgG anti-immunoglobulin (anti-Ig), since it is known that Fab'2 anti-Ig is mitogenic while IgG anti-Ig is not, owing to a dominant inhibitory effect of crosslinking the B cell Fc gamma RII to membrane Ig. While mitogenic doses of Fab'2 anti-Ig induce large and rapid increases in Egr-1 expression, IgG anti-Ig results in smaller increases in Egr-1 mRNA, comparable to that seen with submitogenic concentrations of Fab'2 anti-Ig. However, the correlation between Egr-1 expression and B cell proliferation breaks down when IL-4 is added as a co mitogen to induce B cell proliferation with IgG anti-Ig or submitogenic concentrations of Fab'2 anti-Ig. No corresponding increases in Egr-1 mRNA levels are observed when IL-4 is added. Therefore, IL-4 overcomes Fc receptor-mediated inhibition of B cell proliferation without affecting inhibition of Egr-1 mRNA induction, as demonstrated earlier for c-myc mRNA in this system. PMID- 8232341 TI - Detection of streptococcal pyrogenic exotoxin genes by a nested polymerase chain reaction. AB - Severe invasive disease associated with group A Streptococcus (GAS) has recently increased in frequency. Isolates of GAS from normally sterile sites were examined for the streptococcal pyrogenic exotoxin genes spe A, spe B and spe C to determine if they play a role in this disease. Four primers for each gene were used in a nested polymerase chain reaction (PCR) configuration. The first PCR generated fragments of 818, 1106, and 801 bp, respectively, for the extotoxin genes. The second PCR generated fragments of 500, 912 and 654 bp for the spe A, spe B and spe C genes using the fragments from the first PCR as template. Of 62 strains tested, 35 (56%) contained the spe A gene, and 17 (27%) contained the spe C gene. All GAS strains studied, regardless of disease association, contained the spe B gene. These data corroborate accumulating evidence that the genes encoding pyrogenic exotoxin types B and C are not associated with severe invasive streptococcal illness including streptococcal toxic shock-like syndrome. This PCR based gene detection system has clinical and epidemiologic applications because of its ease of performance, non-isotope labelling, high specificity and sensitivity, and lack of requirement for purified DNA. PMID- 8232342 TI - DNA probes for the detection of Fasciola hepatica in snails. AB - Fasciola hepatica, also called the large liver fluke, is a trematode which can infect most mammals. Monitoring the infection rate of snails, which function as intermediate hosts and harbour larval stages of F. hepatica, is an important component of epidemiological studies on fascioliasis. For this purpose, DNA probes were generated which can be used for the detection of F. hepatica larvae in snails. Four highly repetitive DNA fragments were cloned in a plasmid vector and tested by Southern blot hybridization to the DNA of various trematodes for specificity and sensitivity. The probes Fhr-I, Fhr-II and Fhr-III hybridized only to F. hepatica DNA. Fhr-IV contained ribosomal RNA gene sequences and cross hybridize with the DNA from various other trematode species. Squash blot analysis showed that the different probes were able to detect the parasite larvae in trematode-infected snails even as isolated single larvae. No signals were obtained in squash blots of uninfected snails. Probes Fhr-I, Fhr-II and Fhr-III are thus useful specific tools for studying the epidemiology of fascioliasis. The probe Fhr-IV, because of its broader spectrum, can be used to detect the larvae of a wide range of trematode species of waterbirds, which are the causative agents of swimmer's itch. PMID- 8232343 TI - Single-stranded RNA probes generated from PCR-derived DNA templates. AB - The following report outlines the use of the polymerase chain reaction (PCR) in combination with in vitro transcription to generate single-stranded radiolabelled RNA probes useful for nuclease protection and in situ hybridization experiments. Specific DNA fragments with bacteriophage promoter (T3 and/or T7) sequences at the 5' or 3' end are generated by repeated rounds of amplification. Following purification, these PCR-generated DNA products are used as templates for in vitro transcription with the correct DNA-dependent RNA polymerase. The resultant radiolabelled, single-stranded RNA (ssRNA) can be used for in situ hybridization, Southern or Northern blot analysis, and ribonuclease protection experiments. Sub cloning or hydrolysis of large fragments is not required. Probes can be made from virtually any sequence using a variety of template sources. PMID- 8232344 TI - Determination of bovine rotavirus G and P serotypes by polymerase chain reaction. AB - Among bovine rotaviruses there are two major G serotypes (G6 and G10) and three P serotypes (P1, P5, and P11, each of which is defined on the basis of the VP4 antigenic specificity of NCDV, UK and KK-3, respectively). The nucleotide sequence of a P11 gene (KK-3) was determined. The predicted KK-3 VP4 contained 772 amino-acids and showed 96% amino-acid identity with B223 VP4, an American prototype of P11 bovine rotavirus. Comparative analysis of the genes determining the G and P serotypes of bovine rotaviruses allowed us to develop polymerase chain reaction (PCR)-based assays which distinguished two G serotypes and three P serotypes commonly found in bovine rotaviruses. For determination of G serotypes, a 1013 bp fragment of the VP7 gene was first reverse-transcribed and then amplified with a pair of generic primers. In a second PCR amplification, the 5' generic primer and two different typing primers (either G6- or G10-specific) were used to generate fragments whose sizes served to identify the G serotype. Similarly, for determination of P serotypes, an 864 bp fragment of the VP4 gene was first reverse-transcribed and then amplified with another pair of generic primers. In a second PCR amplification, the 5' generic primer and three different typing primers, each one specific to one of the three P serotypes, were used to generate fragments whose sizes served to identify the P serotype. These assays were able to identify the G and P serotypes of six reference bovine rotavirus strains. PMID- 8232345 TI - A PCR approach to discriminate between integrated and episomal HPV DNA in small clinical specimens. AB - HPV infection has long been implicated in the development of cervical carcinoma. There is strong evidence for association of high-risk HPV types 16 and 18 with cervical intraepithelial neoplasia (CIN) grades 2 and 3, and integration of viral DNA of these types into the host genome has been suggested to play an important role in progression to invasive cervical carcinoma. However, the existing techniques for detection of integrated DNA in clinical specimens are time consuming and require large amounts of template DNA, often unavailable for small premalignant CIN lesions. In this study, a novel, two stage PCR assay was designed to discriminate between integrated and episomal HPV 16 DNA. The first stage was designed to determine whether intact HPV genomes were present. This initial PCR analysis of the entire viral genome in eight segments was successfully applied to authentic human cervical cancers. The second stage consisted of an ANCHOR-PCR-based analysis, developed specifically to discriminate between integrated and episomal HPV DNA, that was successfully tested with cloned HPV containing plasmids used to mimic both episomal and integrated viral DNA. Further optimization and validation will be required for application of the second stage to clinical specimens. The entire assay was developed to be applicable to small colposcopic biopsies or cervical scrape samples, representative of those acquired in routine clinical investigation of CIN 2 or CIN 3, in which determination of the physical state of HPV DNA may provide prognostically valuable information. PMID- 8232346 TI - Variability of the random amplified polymorphic DNA assay among thermal cyclers, and effects of primer and DNA concentration. AB - The reproducibility of the generation of random amplified polymorphic DNA fragments from three commonly used thermal cyclers was determined using identical assay conditions. In all cases, different results were obtained from the three instruments. Variation in the length of the primer (20 nt or 10 nt) did not have any effect on the reproducibility of the assays from the three machines tested. A DNA concentration of 1 ng generated poorly staining DNA fragments whereas concentrations between 10 ng and 100 ng gave similar banding patterns when using the same thermal cycler. Low concentrations of primer (0.05 microM) did not produce any detectable DNA fragments. Increased primer concentrations of 0.25 microM or higher generated intensely staining DNA fragments, and concentrations above 0.5 microM did not improve the clarity of the banding patterns but did direct the synthesis of increasing amounts of very short DNA fragments. Surprisingly, the 20 nt-long primer was able to direct the synthesis of more DNA fragments than a primer of only 10 nt long. PMID- 8232347 TI - Thermonuclease gene as a target nucleotide sequence for specific recognition of Staphylococcus aureus. AB - DNA fragments, 450 bp in length, were amplified by polymerase chain reaction (PCR) from the thermonuclease gene (nuc) carried by seven epidemiologically independent Staphylococcus aureus isolates. Sequencing of the PCR products led us to characterize 210 bp strictly conserved. A 186 bp piece from within this conserved region was cloned into pUC18. The resulting recombinant plasmid, pIP1608, was used as a probe against the cellular DNA of 360 staphylococcal isolates belonging to 28 species. Only the 146 S. aureus isolates, including four which were not thermonuclease producers, had DNA that hybridized with pIP1608. Among the 214 non-S. aureus staphylococci, 55 exhibited a thermonuclease activity. For 32 of these, the enzymatic activity was inhibited by a commercially available polyclonal antiserum directed against the thermonuclease of an S. aureus strain. These results are in favour of the use of pIP1608 as a probe to specifically recognize S. aureus. Furthermore, we propose a method based on colony blot hybridization and potentially useful to enumerate S. aureus cells in biological samples. PMID- 8232348 TI - Fluorescent approaches to diagnosis of Lesch-Nyhan syndrome and quantitative analysis of carrier status. AB - Lesch-Nyhan syndrome is an X-linked recessive disorder caused by molecular defects within the HPRT gene. Deletional forms of this syndrome, most of which are inherited, account for 15% of the cases. In addition, a large percentage of cases are due to de novo point mutations. We have used complementary fluorescence based PCR assays to analyse disease-causing mutations in three unrelated families: (1) inheritance of dye-labelled PCR products of linked polymorphic loci mapping within and flanking the HPRT gene; (2) dye-labelled exon dosage analysis and (3) automated fluorescence-based DNA sequence analysis. Our results using fluorescent, dye-tagged PCR products show that inheritance of two polymorphic small tandem repeats, HPRTB [AGAT]n, mapping within intron 3 of the HPRT gene, and the CA-repeat at DXS294 can be used to establish linkage to the disease. In addition, we modified a previously described PCR protocol to use fluorescent dye labelled oligoprimers and an ABI Gene Scanner in order to rapidly quantitate deletional forms of Lesch-Nyhan syndrome. Quantitative PCR analysis of individual exons followed by dosage analysis confirmed a deletion encompassing exon 9. A similar approach was used to confirm a previously described HPRT gene duplication involving exons 2 and 3. In this analysis, we co-amplified the HPRTB [AGAT]n and HUMARA [AGC]n repeats and confirmed increased exon dosage in carriers for the duplication. DNA sequence analysis remains the method of choice for delineating new disease-causing mutations, most of which are non-deletional forms of Lesch Nyhan syndrome. We have also used a cycle-sequencing strategy employing dye labelled dideoxy terminators and a laser-activated, fluorescence-emission DNA sequencer in order to define carrier status in 10 family members at risk for Lesch-Nyhan syndrome due to a splice donor mutation in intron 7. Our DNA sequence analyses corroborate small tandem repeat (STR) inheritance patterns in this family. Multiple fluorescence-based strategies should facilitate rapid diagnosis of the various Lesch-Nyhan disease-causing mutations. PMID- 8232349 TI - The parental origin of hydatidiform moles and blighted ova: molecular probing with hypervariable DNA polymorphisms. AB - The androgenetic origin of hydatidiform moles, due to a monospermic or dispermic mechanism, has been reported, and a possible pathogenetic relation with blighted ova suggested. To evaluate the origin of hydatidiform moles and their genetic relationship with blighted ova we investigated a series of samples, utilizing several hypervariable DNA polymorphisms by Southern blotting or PCR. Seven complete or partial hydatidiform mole and 49 blighted ovum cases were investigated. The results confirm the androgenetic origin of complete hydatidiform moles, which were always due in our sample to a monospermic mechanism. Our data exclude a relationship between hydatidiform moles and blighted ova, as in the latter a mixed paternal and maternal DNA contribution was always shown. A high incidence of chromosomal abnormalities in blighted ova was also found. PMID- 8232350 TI - Rapid genomic typing of BK virus directly from clinical specimens. AB - A simple and rapid method, PCR-restriction enzyme analysis (PCR-RE) for BK virus (BKV) typing was developed, based on the presence of type-specific restriction enzyme sites in a 327 bp PCR-generated fragment which partially encodes the VP1 protein. The enzymes, Alu I, Xmn I and Ava II, were used to digest the PCR products in two stages. Ethidium bromide banding patterns characteristic of each of four subtypes of BKV were visualized through gel electrophoresis. A total of 37 samples from clinical specimens and culture fluids were successfully subtyped using the PCR-RE. A second method, PCR-sequencing, was applied to samples that generated less than 100 ng of DNA. These were subjected to a second round of PCR and then sequenced from single stranded templates immobilised via a biotinylated primer. The subtypes were assigned on the basis of the type-specific sequences previously characterized. PMID- 8232351 TI - The subthalamic nucleus: a possible target for stereotaxic surgery in Parkinson's disease. AB - Hyperactivity in the subthalamic nucleus (STN) projections to the globus pallidus medialis (GPM) has been established as a crucial feature of parkinsonism in animal models of Parkinson's disease (PD). Recent experiments blocking the STN glutamatergic output to GPM or lesioning the STN support this concept by showing a dramatic reversal of parkinsonism. We review the role of stereotaxic surgery in the past and the possible future application of subthalamotomy for PD. PMID- 8232352 TI - Pre- and postcentral cortical somatosensory evoked potentials in hemiparkinsonism. AB - We recorded cortical frontal, central, and parietal somatosensory evoked potentials (SEPs) in 9 patients with hemiparkinsonism and in a group of 25 healthy volunteers. No differences were observed in the SEPs recorded after stimulation of the asymptomatic and symptomatic sides in the patients. Likewise, comparison with the healthy controls did not reveal significant group differences or abnormal waveforms in the patients. Even frontal N30 deflection, which has been reported to be diminished in Parkinson's disease (PD), was normal and symmetric in the patients. Therefore, no evidence was found for altered sensory input to the motor or premotor and supplementary motor cortices in PD. PMID- 8232353 TI - Increased striatal glucose consumption in Sydenham's chorea. AB - Positron emission tomography and 18F-fluorodeoxyglucose were used to measure the regional cerebral glucose consumption in a 15-year-old choreatic girl with classical Sydenham's chorea shortly after the onset of hyperkinetic movements and 5 months later after chorea had resolved and in a 74-year-old hemichoreatic woman with long-standing hyperkinesia as a residuum of Sydenham's chorea in adolescence. Whereas cerebellar, thalamic, and cortical glucose consumption was within normal limits in both patients, lentiform and caudate glucose consumption was significantly increased in both hemispheres of the 15-year-old patient and in the hemisphere contralateral to the chorea in the 74-year-old patient. In the younger patient, striatal glucose consumption returned to normal after her hyperkinesia had disappeared with antibiotic therapy. The observation of an increase in striatal glucose consumption in Sydenham's chorea, in contrast to the decrease of this variable encountered in the vast majority of other choreatic disorders, leads to questioning the pathophysiology of chorea in humans and suggests the use of emission tomographic measurement of variables related to cerebral energy metabolism for differential diagnosis in choreatic disorders. PMID- 8232354 TI - Correlation of clinical response in apomorphine test with D2-receptor status as demonstrated by 123I IBZM-SPECT. AB - The knowledge of functional capacities of postsynaptic dopaminergic receptors in parkinsonian syndromes is important for differential diagnosis and for planning therapeutic approaches. Subcutaneous apomorphine challenges serve as a pharmacological tool in testing dopaminergic responsiveness, but discrepancies between results of the apomorphine test and long-term levodopa treatment remain. 123I IBZM (I-123 labeled iodobenzamide) as a dopaminergic receptor ligand allows depiction of D2-receptors by means of SPECT methods. The correlation between dopaminergic responsiveness and D2-receptor status as demonstrated by 123I IBZM SPECT imaging was assessed by applying an apomorphine test to 41 patients with parkinsonian syndromes. All subsequently underwent an 123I IBZM-SPECT. Apomorphine responders showed a significantly higher binding of 123I IBZM than nonresponders, and patients with idiopathic Parkinson's disease (IPD) had higher D2-receptor density as visualized by SPECT than patients with other parkinsonian syndromes. The marked overlap between the groups allowed a reliable prediction only in patients with an abnormally low basal ganglia/frontal cortex ratio of 123I IBZM binding. PMID- 8232355 TI - Relief of akinesia by apomorphine and cerebral metabolic changes in Parkinson's disease. AB - The cerebral metabolic rate of glucose was measured in 14 Parkinson's disease patients with severe on-off fluctuations. Two positron emission tomography (PET) scans with [18F]fluorodeoxyglucose were performed, one after a challenge of subcutaneous apomorphine at a dose able to relieve akinesia within 15 min and the other with the vehicle. Apomorphine reduced glucose utilization by 4-6% in the lenticular nuclei and the occipital cortex and by 6-9% in the thalamic nuclei, but this effect was not statistically significant. Thus, central stimulation of dopamine receptors by apomorphine in advanced Parkinson's disease is not associated with cerebral metabolic changes as assessed by PET. Despite a dramatic improvement of the motor state, the global neuronal activity in the striatum and its downstream projections remains stable, suggesting an equilibrium between excitatory and inhibitory dopaminergic activities. PMID- 8232356 TI - Reliability of the Columbia scale for assessing signs of Parkinson's disease. AB - Inter- and intrarater reliability in scoring the signs of Parkinson's disease using the original Columbia scale and a modified version of this, the Sydney scale, were assessed in five neurologists participating in a long-term study of Parkinson's disease. Scoring was done on video recordings of 41 patients whose disability ranged from mild to severe. Although all the neurologists were familiar with the scales and had received training designed to produce uniformity of scoring, interrater reliability was poor. The mean score for the Columbia scale varied from 18.6 to 30 and for the Sydney scale from 15.2 to 23.2. By contrast, intrarater reliability was good. This study highlights the limitations of clinical rating scales in Parkinson's disease when more than one rater is used. In designing clinical trials, every effort should be made to ensure that the same patient is always assessed by the same rater. PMID- 8232357 TI - Use of botulinum toxin type F injections to treat torticollis in patients with immunity to botulinum toxin type A. AB - Fifteen patients with torticollis who had been treated with repeated injections of botulinum toxin type A (botox A) developed antibodies to the toxin. This resulted in loss of benefit in the 13 patients who had improved with botox A injections and failure to develop muscle atrophy after injection in all 15 patients. Patients were then injected with botulinum toxin type F (botox F) in the same muscles that had been injected with botox A. Ten of the 15 improved after botox F injections, including 9 of the 12 patients who had improved with type A toxin. Six of 9 patients with pain had improvement in pain after botox F injections. Patients reported similar improvement with type F and type A toxins, but duration of benefit was approximately 3 months with type A and approximately 1 month with type F. Botox F is an effective treatment for torticollis in patients who are immune to botox A. The usefulness of type F toxin, however, is limited by short duration of benefit. PMID- 8232358 TI - Therapeutic trial of milacemide in patients with myoclonus and other intractable movement disorders. AB - We performed a therapeutic trial with the glycine precursor, milacemide, on 10 patients with intractable movement disorders. Six had myoclonus of various etiologies and one each had progressive supranuclear palsy, Filipino X-linked dystonia with parkinsonism, painful legs and moving toes, and stiff-person syndrome. Milacemide was initiated at a dose of 2,400 mg/day, orally, and increased gradually to a maximum of 4,800 mg/day. No clear-cut observable improvement occurred. There were no serious adverse effects. PMID- 8232359 TI - Adult onset familial cervical dystonia: report of a family including monozygotic twins. AB - We report the first family with adult onset cervical dystonia in which monozygotic twins and multiple family members are affected. In this family, the disease exhibits an autosomal dominant inheritance pattern, and all affected members have only cervical dystonia. Five members have definite cervical dystonia, and five others have possible cervical dystonia. Although identical genotypically, the twins demonstrate some phenotypic variation. Despite long follow-up, no family members have shown progression from focal to generalized dystonia. This family may prove valuable in identifying a gene locus for cervical dystonia and hence determine whether a single gene locus exists for hereditable focal dystonia and generalized dystonia. PMID- 8232360 TI - Blink reflex and the masseter inhibitory reflex in patients with dystonia. AB - The excitatory and inhibitory interneuronal pathways in the brainstem are tested by examining the blink reflex and the masseter inhibitory reflex, respectively. We studied the R2 component of the blink reflex and the SP2 component of the masseter inhibitory reflex and their recovery cycle in 56 patients with various forms of dystonia. In patients with cranial, cervical, and generalized dystonia, but not in patients with extracranial segmental dystonia, the recovery cycle of both reflexes was enhanced. The recovery cycle of R2 and SP2 can demonstrate subclinical changes in excitability of brainstem interneurons. The degree of enhancement of the recovery cycles did not correlate, however, with the severity of clinical facial muscle impairment. In addition, the recovery cycles correlated positively with each other, showing that excitatory as well as inhibitory interneuronal pathways in the brainstem are perturbed in dystonia. Study of the trigemino-facial and trigemino-trigeminal reflexes provides an objective tool for assessing functional abnormalities in dystonia. PMID- 8232361 TI - Anxiety and motor performance in Parkinson's disease. AB - Previous work has suggested that anxiety disorders are common in patients with Parkinson's disease. To study the relationship between anxiety and response fluctuations in Parkinson's patients further, we compared changes in Spielberger anxiety state scores with changes in parkinsonian disability as determined by the Parkinson's symptom diary (PSD). Anxiety state and PSD scores were obtained in 19 patients with idiopathic Parkinson's disease during on and off periods. Spielberger anxiety state scores were higher during off periods than during on periods (38.8 +/- 12.4 vs. 45.6 +/- 12.4, p < 0.03); further, the magnitude of the change in anxiety state scores was correlated with the change in PSD scores (rs = 0.616, p = 0.006). Spielberger anxiety trait scores were also correlated with disease duration. Our findings support existing data suggesting that anxiety can contribute significantly to morbidity in Parkinson's disease and suggest that anxiety varies with fluctuations in motor performance. PMID- 8232362 TI - Relationship between osteopenia and clinical characteristics of Parkinson's disease. AB - Pathological bone changes affect locomotor activity and may influence the outcome and prognosis of Parkinson's disease (PD). In this study, we determined the relationship between bone changes and PD. Bone study was performed by multiple scanning x-ray photodensitometry (MD/MS) in 64 patients with PD and 42 age matched controls. We then compared the results with the clinical characteristics of PD. Osteopenia was detected in 22 (53.6%) of the 41 female and 6 (26%) of the 23 male patients, and in 6 (26%) of the 23 female and 2 (10.5%) of the 19 male controls. The frequency of osteopenia was significantly greater in the female patients than in the male patients or the female controls. Osteopenia was related to the duration of PD in the men, but not in the women. Twenty of 40 PD patients' hands showed side-related differences in the analysis of both hands. In 19 of the 20 patients, the side of more severe osteopenia coincided with that of parkinsonian symptoms, suggesting that osteopenia is related to the pathophysiology of PD. PMID- 8232363 TI - Orobuccal dyskinesia associated with trihexyphenidyl therapy in a patient with Parkinson's disease. AB - Dyskinesia is a common adverse effect complicating chronic dopaminergic therapy for Parkinson's disease. Movements are frequently choreic in nature and have been ascribed to overstimulation of "supersensitive" striatal postsynaptic dopamine receptors. Anticholinergic medications, despite some clinical efficacy in Parkinson's disease, have rarely been reported to cause dyskinesia. We report a patient with Parkinson's disease who developed orobuccal dyskinesia while being treated with trihexyphenidyl (Artane). Dyskinesia was observed following the introduction of trihexyphenidyl, resolved with its discontinuation, and reappeared with its reinstitution. Carbidopa-levodopa (Sinemet) alone did not cause dyskinesia but augmented dyskinesia associated with trihexyphenidyl. PMID- 8232364 TI - A case of symptomatic hemidystonia improved by ventroposterolateral thalamic electrostimulation. AB - A 16-year-old boy presented with a left anterothalamic lesion secondary to an extradural hematoma. A few months later, a right hemidystonia developed. The dystonic posture, which predominated in the right hand, was not associated with any motor deficit or sensory loss. Superficial sensory stimulation of the right palm and forearm and proprioceptive stimuli induced by passive movements of the right thumb decreased the dystonic posture. Electrical stimulation of the left ventroposterolateral nucleus of the thalamus induced a dramatic improvement in the dystonic postures and movement of the upper right limb. This finding suggests that the role of tactile and proprioceptive stimulation should be analyzed in patients presenting with a symptomatic hemidystonia. When such sensory stimulation is effective, thalamic stimulation may be tried in patients whose condition is incapacitating. PMID- 8232365 TI - Effects of thalamic stimulation on tremor, balance, and step initiation: a single subject study. AB - This study was conducted to evaluate the clinically apparent balance improvements in a patient with Parkinson's disease who had stimulating electrodes surgically implanted to the VIM nucleus of the right thalamus for control of left-upper extremity tremor. Experiments were conducted to determine if balance improved simply because the large-amplitude upper-extremity tremor was reduced or if the neural control of balance improved. Using EMGs and forceplate recordings, we quantified the effects of the thalamic stimulation on the contralateral upper extremity tremor and on the lower-extremity postural muscle activations for quiet stance, step initiation, and equilibrium responses to surface displacements. The results demonstrated that, beside reducing the amplitude and destabilizing effects of the upper-extremity tremor, the thalamic stimulation was also effective in reducing tremor activity of the trunk and contralateral lower extremity muscles. In addition, the contralateral lower-extremity muscle activation patterns, strengths, and durations for the balance tasks were enhanced during stimulation. These results suggest that thalamic stimulation improved this patient's balance by reducing tremor in the contralateral extremities and by increasing burst duration and magnitude of the tibialis anterior, which functions as the postural prime mover for the step initiation and balance tasks. PMID- 8232366 TI - What is it? Case 1, 1993: parkinsonism, dysautonomia, and ophthalmoparesis. PMID- 8232367 TI - The architecture and dynamics of developing mind: experiential structuralism as a frame for unifying cognitive developmental theories. AB - This Monograph presents a theory of cognitive development. The theory argues that the mind develops across three fronts. The first refers to a general processing system that defines the general potentials of mind to develop cognitive strategies and skills. The second refers to a hypercognitive system that governs self-understanding and self-regulation. The third involves a set of specialized structural systems (SSSs) that are responsible for the representation and processing of different reality domains. There are specific forces that are responsible for this organization of mind. These are expressed in the Monograph in terms of a set of five organizational principles. The developmental course of the major systems is outlined. Developmental change is ascribed by the theory to the interaction between the various systems. Different types of development require different change mechanisms. Five studies are presented that provide empirical support for these postulates. Study 1 demonstrated the organizational power of principles and SSSs. Study 2 showed that the SSSs constrain the effect of learning. Study 3 established that the hypercognitive system does function as the interface between tasks and SSS-specific processes or between SSSs and general cognitive functions such as attention and memory. Study 4 investigated the relations between one of the components of the processing system, storage, and two different SSSs expressed via two different symbolic systems, namely, the numeric and the imaginal. Finally, Study 5 examined the interaction between the components of the processing system and the relations between each of these components and one SSS, namely, the quantitative-relational SSS. The theoretical implications of these studies with regard to general issues, such as the nature of representation, the causation of cognitive change, and individual differences in cognitive development, are discussed in the concluding chapter. PMID- 8232368 TI - Free-living amebas of the genera Acanthamoeba and Naegleria: an overview and basic microbiologic correlates. AB - Free-living amebas of the genera Acanthamoeba and Naegleria are allied in basic cell biology, ecology, and human disease-producing potential. However, several enigmas surrounding these amebas need further intellectual and scientific scrutiny. For instance, a clearer differentiation is needed of factors delineating pathogenic species--those most frequently associated with human infections--from nonpathogenic species. Further, have the pathogenic species bridged the gap in a step-wise fashion between their habitat and the human host to express their disease-producing potential? Second, what attributes of amebas account for the spectrum of disease caused by Acanthamoeba and Naegleria? In the brain, Naegleria is highly destructive of tissue in the course of a rapidly evolving, hemorrhagic primary meningoencephalitis in normal individuals. Central nervous system invasion by Acanthamoeba, however, occurs only in compromised hosts, and is a more slowly evolving subacute to chronic encephalitis. Further, the epidemiology of the infections are disparate. Naegleria is acquired from exposure to contaminated fresh water; this epidemiologic link is absent in Acanthamoeba meningoencephalitis. Naegleria invades the central nervous system via the olfactory nerve; Acanthamoeba is deposited in the central nervous system via hematogenous spread from a pulmonary or cutaneous focus. Additionally, Naegleria is apparently restricted to the brain; Acanthamoeba is not so bridled, and invades more sites in the human body. Finally, Acanthamoeba are beginning to appear opportunistically more frequently in patients infected with the human immunodeficiency virus. To avoid the same lack of effective therapeutics we still face in treating cryptosporidium infections in these patients, more emphasis must be placed on clinical trials dealing with the management of Acanthamoeba infections in patients with AIDS. PMID- 8232369 TI - Free-living amebas: infection of the central nervous system. AB - Pathogenic free-living amebas of the genera Naegleria and Acanthamoeba and the leptomyxid ameba of the order Leptomyxida may be capable of producing disease in the central nervous system of human beings and animals. These amebas are distributed worldwide in thermally polluted streams, coastal and fresh water, dust, soil, and sewage, and heating, ventilating, and air conditioning units. N. fowleri may produce primary amebic meningoencephalitis, a rapidly fatal central nervous system infection. By contrast, the Acanthamoeba spp. and the recently described leptomyxid ameba may produce granulomatous amebic encephalitis, a protracted central nervous system disease, usually in immunocompromised hosts. The leptomyxid ameba may produce clinical symptoms similar to and histopathologic features almost identical to those in GAE. Contact-lens wearers may also develop Acanthamoeba keratitis, chronic ulceration of the corneal epithelium due to Acanthamoeba spp. The various central nervous system diseases produced by these free-living amebas result in divergent epidemiological patterns, diverse clinical manifestations, and distinct pathological features, and require different treatment. PMID- 8232370 TI - Acanthamoeba keratitis: there and back again. PMID- 8232371 TI - Epidemiology of infections with free-living amebas and laboratory diagnosis of microsporidiosis. AB - Acanthamoeba, leptomyxid ameba, and microsporidia have recently been recognized as organisms that cause opportunistic infections in patients with HIV and AIDS. As the numbers of patients with HIV and AIDS increase, more cases of amebic encephalitis and microsporidiosis are likely. PMID- 8232372 TI - Stroke prevention in atrial fibrillation. AB - Atrial fibrillation (AF) is a risk factor for ischemic stroke. In randomized trials, AF raised the risk of stroke nearly sixfold, cumulating in a 35% risk over a lifetime. Anticoagulation with warfarin reduces the danger of ischemic stroke, but carries hemorrhagic risks, making this agent unsuitable for treating many patients. Platelet inhibitor therapy with aspirin was highly effective for patients younger than 75 years of age in one study, but the reason for lower efficacy in older individuals is perplexing. These trials support a thrombotic mechanism for most strokes in patients with AF, but leave physicians in a quandary as to selection of optimum prophylaxis. Secondary analysis of patients given placebo identified predictors of thromboembolism, including a history of hypertension, congestive heart failure, and prior stroke or transient ischemic attack, and echocardiographic findings of left ventricular dysfunction or left atrial enlargement. The absence of these risk factors selects a fairly large subgroup of AF patients at comparatively low risk of stroke, for whom the danger and inconvenience of chronic anticoagulation may not be warranted. It is becoming clear that specific clinical and echocardiographic features allow individualized antithrombotic approaches within the broad category of patients with AF, to enhance therapeutic benefit while minimizing hemorrhagic risk. PMID- 8232373 TI - Frontiers in the treatment of ischemic stroke. AB - Advances in the understanding of the pathophysiology of ischemic neuronal death have led to the development of new approaches to treating acute stroke. Concurrently technologic advances have permitted application of older approaches in a more sophisticated manner. These advances herald an era in which rapid, precise evaluation and treatment of the patient with acute stroke may dramatically alter prognosis. This article reviews some of these advances. PMID- 8232374 TI - Urologic aspects of prevention of end-stage renal disease in spinal cord injury. PMID- 8232375 TI - Pathophysiology of end-stage renal disease in spinal cord injury. PMID- 8232376 TI - Obesity surgery: dietary and psychosocial expectations and reality. AB - Psychosocial and dietary habits have been compared in patients considering (Pre n = 33) or who had (Post n = 32) surgery for morbid obesity. Failure of conventional diets was attributed to lack of self-discipline (Pre 92% [22/24] vs. Post 87% [20/23], NS). Consumption of fast foods fell (Pre 70% [23/33] vs. Post 16% [5/32], p < 0.0001). Preoperative patients had unrealistic social expectations. They exaggerated the prospect of improved friendship (Pre 67% [22/32] vs. Post 34% [11/21], p < 0.05), erroneously anticipated better sex (Pre 78% [25/32] vs. Post 50% [15/30], p < or = 0.05), predicted better acceptance at work (Pre 85% [23/27] vs. Post 50% [15/30], p < or = 0.05), and misanticipated improved relationship with their partner (Pre 77% [20/26] vs. Post [47% 8/17], p < or = 0.05). Two factors predicted becoming employed following surgery: age (became employed [n = 5] 28 +/- 2 years vs. remained unemployed [n = 12] 44 +/- 4 years, p < 0.05) and percentage of excess weight lost (became employed 76 +/- 11 vs. remained unemployed 51 +/- 7, p < 0.05). The free support group was "useful" (17/17), yet only 5% attended regularly. Patients considering obesity surgery had specific unrealistic psychosocial expectations. They infrequently availed themselves of postoperative professional help. We identify the features associated with gaining employment. PMID- 8232377 TI - Phase II trial of etoposide, carboplatin, and ifosfamide as salvage therapy in advanced ovarian carcinoma. AB - A phase II study combining etoposide with carboplatin and ifosfamide as salvage therapy in advanced ovarian cancer was undertaken. Objective responses were achieved in 37.5% of 16 evaluable patients with a mean progression-free interval of 8.6 months. Stable disease was present in 25% of patients; in 37.5% of patients the disease progressed on salvage. Based on original response to front line therapy, patients were classified as being platinum-sensitive (group I) or platinum-refractory (group II). Clinical response to salvage therapy was seen in 44.5% of group I patients, but in only 28.6% of group II patients. This difference was not statistically significant. When a more precise definition of platinum sensitivity was applied, clinical responses were seen in 54.5% of group I patients, but no responses were noted among group II patients (p < 0.05). Platinum sensitivity appeared to be an important factor in achieving a response with this regimen. This combination was well tolerated, myelotoxicity being the dose-limiting toxicity encountered. No life-threatening, nonhematologic toxicities were seen. One death occurred secondary to nadir sepsis. The combination of etoposide, carboplatin, and ifosfamide is an active salvage regimen in patients with advanced ovarian carcinoma; however, severe myelotoxicities and inability to produce long-term responses underscore the need for continued trials to find a more durable salvage regimen. PMID- 8232378 TI - The enteroinsular axis and endocrine pancreatic function in chronic alcohol consumers: evidence for early beta-cell hypofunction. AB - Chronic alcohol consumers may have, as judged by functional criteria, exocrine as well as endocrine pancreatic dysfunction, the latter represented by a decreased insulin response to an oral glucose load. To investigate whether this decreased insulin response was due to an ethanol-induced beta-cell dysfunction or to an ethanol-induced dysfunction of the enteroinsular axis, we determined glucose, insulin, and C-peptide plasma concentrations following an oral and an intravenous glucose load in 16 healthy volunteer nonalcohol consumers and in 10 chronic alcohol consumers. In each group, total integrated response for glucose did not significantly change whether glucose was given orally or intravenously, indicating isoglycemic glucose loads. The total integrated response values for insulin in the alcoholic group following both glucose loads as well as C-peptide plasma concentrations were significantly lower than in the control group. Moreover, in both groups the insulin TIR values following the oral glucose load were significantly greater than the values obtained following the intravenous glucose load, indicating an incretin effect. These results indicate that the decreased insulin response observed in alcoholics was not caused by a dysfunction of the enteroinsular axis because it also occurred following an intravenous glucose load, but by an ethanol-induced beta-cell dysfunction because C-peptide and insulin were proportionally decreased in this group. PMID- 8232379 TI - Thoracic herniated discs: review of the literature and 12 cases. AB - In comparison with herniations of lumbar or cervical intervertebral discs, symptomatic thoracic disc herniation is rare. Between 1986 and 1991, 12 cases of thoracic herniated discs were treated at The Mount Sinai Hospital, New York City. Most patients had back pain or myelopathy. Nine of the disc herniations occurred at the lowest six thoracic interspaces. Eight patients underwent costotransversectomy and discectomy. Seven of these patients improved without complication. One patient was subsequently found to have a spinal arteriovenous malformation below an incidental herniation that had been identified by magnetic resonance imaging. Because of the nonspecificity of the signs and symptoms, as well as the prevalence of incidental herniations on imaging, a careful clinical and radiologic correlation is mandatory when diagnosing this uncommon pathology. PMID- 8232380 TI - Screening for human immunodeficiency virus and sexually transmitted diseases in an inner-city colposcopy clinic. AB - Among patients attending an inner-city colposcopy clinic, the prevalence of Chlamydia trachomatis was 22/375 (5.0%), Neisseria gonorrhoeae, 3/375 (0.8%), and seropositivity for syphilis, 10/375 (2.7%). In addition, 13/261 (5.0%) of asymptomatic women agreeing to voluntary human immunodeficiency virus (HIV-1) antibody screening were HIV-1 seropositive. Our data support incorporating screening for and education on sexually transmitted diseases and HIV into the work of our colposcopy clinic. PMID- 8232381 TI - Case report: evolution of a type B aortic dissection following renal artery angioplasty. AB - Percutaneous renal artery angioplasty is an accepted treatment for renal artery stenosis. A variety of complications have been reported related to this procedure. The authors report a case of Type B aortic dissection occurring immediately following renal artery angioplasty, a previously unreported complication of renal artery angioplasty. A mechanism for this complication in this patient is postulated. PMID- 8232382 TI - Propofol use in malignant hyperthermia: a case report. PMID- 8232383 TI - Congenital myasthenic syndromes: I. Deficiency and short open-time of the acetylcholine receptor. AB - A 5.5-year-old girl had myasthenic symptoms since birth. Tests for antiacetylcholine receptor (AChR) antibodies were negative. To investigate the character of the neuromuscular transmission defect, an intercostal muscle specimen was obtained at age 27 months. Immune deposits were absent from the endplates. On electron microscopy, most postsynaptic regions appeared normal, but the density of AChR on the junctional folds was diffusely reduced. In vitro microelectrode studies revealed that the number of transmitter quanta released by nerve impulse was normal. The amplitude of miniature of endplate potentials and currents was abnormally low. A study of the kinetic properties of AChR by analysis of acetylcholine-induced current noise demonstrated a significant decrease in mean channel open-time; the mean channel conductance was normal. The safety margin of neuromuscular transmission in this disorder is likely to be compromised by the deficiency and abnormal kinetic properties of AChR. The findings are unique among those patients with congenital AChR deficiency described to date. PMID- 8232384 TI - Congenital myasthenic syndromes: II. Syndrome attributed to abnormal interaction of acetylcholine with its receptor. AB - A 21-year-old woman had myasthenic symptoms since birth that responded poorly to anticholinesterase therapy. Tests for acetylcholine receptor (AChR) antibodies were negative. An intercostal muscle specimen was obtained to investigate the character of the neuromuscular transmission defect. There were no immune deposits at the endplates. The quantal content of the endplate potential was normal. Miniature endplate potentials and currents were very small, but the number of AChR per endplate was normal. On electron microscopy, the synaptic vesicles were of normal size, the junctional folds were intact, and the density and distribution of AChR on the folds was normal. The kinetic properties of AChR were studied by analysis of acetylcholine (ACh)-induced current noise. The mean single channel conductance was normal. The noise power spectrum was abnormal, containing two components of different time course. This could result from an abnormal interaction of ACh with AChR, or from two populations of AChR at the endplate. The second possibility is unlikely because if two populations of AChR were present at the endplate, then both would have to have low conductance to explain the small miniature endplate current, but the average conductance of the channels that did open was normal. PMID- 8232385 TI - F tacheodispersion: quantitative analysis of motor fiber conduction velocities in patients with polyneuropathy. AB - The distribution of motor fiber conduction velocities (FCVs) of the ulnar and peroneal nerves in patients with polyneuropathy was estimated using F tacheodispersion, a technique based on F wave latencies. Twenty-three ulnar and 23 peroneal nerves were studied. Three groups of nerves were identified according to conventional, M response, and F wave measurements: those with all parameters normal (group 1), those with abnormal F wave parameters only (group 2), and those with all parameters abnormal (group 3). Application of F tacheodispersion to nerves in group 1 revealed a shift of the distribution of FCVs toward the lower normal values in 6 of 7 nerves. In group 2, a significant proportion of FCVs were below the lower normal limits despite normal maximum conduction velocity estimated by the M response. In group 3, all nerve fibers were abnormally slow. The results indicate that application of F tacheodispersion in clinical practice will increase sensitivity of nerve conduction studies and enable quantitative estimation of conduction in a motor nerve fiber population. PMID- 8232386 TI - Acute "axonal" Guillain-Barre syndrome in childhood. AB - We identified 5 of 44 consecutive children (11%) with Guillain-Barre syndrome who had electrophysiologic evidence of severe reduction of the mean amplitude of the compound motor action potentials (mean CMAP amplitude < 10% of lower limit of normal). EMG studies revealed profuse fibrillation activity in distal and proximal muscles after 2 weeks of onset. We compared this group with 16 consecutive children with GBS prospectively evaluated over 1 year, all of whom presented a mean CMAP amplitude > 10% of lower limit of normal. Children in the first group were more likely to require assisted ventilation (60% vs. 6.2%) and were more frequently quadriplegic at the peak of their disability (80% vs. 18.7%). They also required longer periods to improve one functional grade (mean 63.6 days vs. 16.6 days) and to become ambulatory (mean 156 days vs. 17.6 days). Moreover, only the children in the first group had distal atrophy of four limbs after 1 year of follow-up. Severe reduction of the mean amplitude of the CMAPs in children with GBS identifies a subgroup of patients with axonal damage that produces more severe weakness and delayed recovery. PMID- 8232387 TI - Beta-adrenergic potentiation of E-C coupling increases force in rat skeletal muscle. AB - We examined the mechanism(s) which allow terbutaline, a beta 2-adrenergic agonist, to increase isometric force in bundles of normal and denervated rat soleus fibers. Terbutaline (10 mumol/L) potentiated tetanic contractions during exposure to 1 mmol/L ouabain, 10 mumol/L nifedipine, or 0.5 mmol/L iodoacetate. Terbutaline induced equivalent increases in submaximal potassium (K+) contracture and tetanic force: these effects were mimicked by 2 mmol/L dibutyryl-cyclic AMP. Therefore, terbutaline increased force by a cyclic AMP-dependent mechanism other than enhancement of sodium-pump activity, dihydropyridine sensitive Ca2+ currents, glycolysis, or action potentials. Pretreatment with 1 mmol/L caffeine induced submaximal potentiation of peak tetanic force but prevented further potentiation by terbutaline. This suggested that terbutaline did not influence the myofilaments, but acted on the sarcoplasmic reticulum (SR) to increase the myoplasmic Ca2+ concentration and hence force production. We speculate that force is potentiated following beta-adrenoceptor activation by a cyclic AMP-dependent phosphorylation of Ca2+ release channels to facilitate SR calcium release during tetanic stimulation. PMID- 8232388 TI - Determinants of mean motor unit size: impact on estimates of motor unit number. AB - The purpose of this study was to compare two fundamentally different methods of deriving the average surface-detected motor unit action potential (S-MUAP) size from which to calculate a motor unit number estimate (MUNE), namely: (1) the simple arithmetic average of S-MUAP parameter values; and (2) a computer-derived datapoint by datapoint average waveform which takes account of differences in S MUAP shapes and durations. Multiple point stimulation was used to collect representative samples of between 11 and 20 S-MUAPs (mean 15 +/- 2 SD) from the median-innervated thenar muscles of 20 healthy control subjects between 20 and 76 years of age (mean 48 +/- 19 SD). The average S-MUAP size based on peak-to-peak amplitude, negative peak amplitude, and negative peak area measurements was calculated using the two different methods. The mean S-MUAP sizes based on the average waveform were significantly lower in all cases than those based on the simple average of S-MUAP parameter values. Differences tended to be greatest for MUNEs based on peak-to-peak amplitude (35%), less for negative peak amplitude (20%), and least for negative peak area (16%). PMID- 8232389 TI - Acetylcholine receptor subunit gene expression in thymic tissue. AB - It is controversial whether acetylcholine receptors (AChR) or AChR-like proteins exist in human thymus. To evaluate this question we isolated RNA from paraffin embedded thymic tissue of 5 myasthenics and 5 nonmyasthenics. RNA was subjected to reverse transcription followed by polymerase chain reaction (RT-PCR) using primers specific for beta-actin, subunits of the fetal- and adult-type AChR, and Myf-4, a gene product which regulates AChR expression in muscle. beta-Actin transcripts were identified in specimens from 8 of 10 patients. In these 8 patients alpha- and epsilon-subunit transcripts were identified. Presence of these transcripts did not correlate with thymic pathology or clinical presentation. No gamma-subunit (specific for the fetal-type AChR) or Myf-4 transcripts were found. Our results indicate that mRNA for subunits of the adult type AChR are expressed in thymic tissue by mechanisms not involving Myf-4. AChR subunits in pathological thymus may provide a target for immune attack in MG. However, as AChR subunits were found in thymus tissue from myasthenics and nonmyasthenics, the presence of AChR subunits in thymus alone is not sufficient to produce myasthenia gravis. PMID- 8232390 TI - The influence of changes in the intensity of magnetic stimulation on coil output. AB - We measured the peak voltage induced in a sensing loop by a Magstim 200 magnetic stimulator. Coil output varied little for repeated stimulation at the same intensity over a wide range of coil output. In contrast, the first stimulus immediately after a change in intensity was of larger amplitude and showed greater variability than subsequent stimuli. The effect was seen for changes in intensity of 5% and 60% and was greater for reductions than for increases in stimulation intensity. Stimulation immediately after a reduction in intensity from 100% to 40% resulted in peak induced voltages as high as those recorded for repeated stimulation at 43% of maximum coil output. Increased coil output following changes in stimulation intensity may affect measurement of the threshold old for motor cortical stimulation. The effect of a change in intensity could be minimized by delaying stimulation for at least 30 s or discarding the first stimulus after the change. PMID- 8232391 TI - A statistical approach to fiber diameter distribution in human sural nerve. AB - Fifty-one normal sural nerve biopsies were obtained from 800 diagnostic biopsies. The external diameter distribution of myelinated fibers was described using the sum of two beta probability density functions, describing the thin as well as the thick fiber group. A cross-sectional study using these distribution functions showed increasing values of the peak of the larger fiber group, the diameter of the thickest fibers, and the separation between the smaller and the larger groups until the beginning of adult life. The transition from a uni- into a bimodal histogram occurred gradually between 7 and 13 months. Total transverse fascicular area increased with age, whereas fiber density decreased significantly with age. The number of fibers remained stable over age. The relative proportion of the numbers of fibers in both groups described by one of the beta distributions remained constant over age. This occurred despite a marked decrease in the number of small fibers with a diameter less than, e.g., 6.5 microns. The results indicated an outgrowth of especially the larger myelinated fibers with age. This process continues with decreasing intensity into adult life. PMID- 8232392 TI - Effect of the inhibitory phenomenon following magnetic stimulation of cortex on brainstem motor neuron excitability and on the cortical control of brainstem reflexes. AB - We studied the effect of the inhibitory phenomenon following transcranial magnetic stimulation (TMS) on brainstem motor neuron excitability and on the cortical control of brainstem reflexes in 5 normal subjects. The R1 component of the blink reflex assessed excitability of the facial nucleus during the period of post-TMS inhibition (experiment 1). TMS effect on the cortical control of brainstem reflexes was assessed by delivery of a stimulus that was less than or equal to the threshold for the R1 with the subject relaxed, yet elicited a potentiated reflex during maneuvers requiring cortical input (i.e., voluntary eye closure and anticipation, experiment 2). In experiment 1, a suprathreshold electric stimulus applied during post-TMS inhibition elicited an R1 response that was preserved in amplitude (mean 476 +/- 44 microV; 123%) compared to control responses not preceded by TMS (mean 386 +/- 31 microV). Conversely, in experiment 2, when the R1 was dependent on cortical drive, the mean R1 amplitude during post TMS inhibition was only 15% (P < 0.01) and 8% (P < 0.01) of control values obtained during eye-closure and anticipation, respectively. These findings imply that the inhibition following TMS originates rostral to the facial nucleus, at a cortical level. PMID- 8232393 TI - Emery-Dreifuss muscular dystrophy with unusual features. AB - Two families with Emery-Dreifuss muscular dystrophy (EMD) are described. Several unusual features for EMD are emphasized. One of the patients had severe neuromuscular disability with inability to walk during early childhood. This patient also had mild bifacial paresis. His brothers had the typical slow progression of EMD. In some of the patients, muscle weakness distribution was more widespread than has usually been reported, with prominent involvement of finger extensors. It is suggested that there is a wide phenotypic spectrum in EMD. In both families, the disease segregated with markers spanning the EMD locus in Xq28. PMID- 8232394 TI - Sensitivity of three median-to-ulnar comparative tests in diagnosis of mild carpal tunnel syndrome. AB - We studied 193 hands of 113 patients referred for typical carpal tunnel syndrome (CTS). Ninety-five (49%) hands had normal median distal motor latency (< or = 4.2 ms) and normal or borderline sensory conduction velocity from digit 2 stimulation (> or = 45 m/s). In these cases we performed three median to ulnar comparative tests: (1) difference between median and ulnar distal motor latencies recorded from the second lumbrical and interossei muscles (2L-INT); (2) difference between median and ulnar sensory latencies from digit 4 stimulation (D4M-D4U); and (3) difference between median and ulnar mixed nerve latencies from palmar stimulation (PM-PU). The 2L-INT difference was > or = 0.6 ms in 10% of hands. PM-PU and D4M D4U were > or = 0.5 ms in 56% and 77% of hands, respectively. The greater sensitivity of D4M-D4U might be explained by the funicular topography and consequent greater susceptibility to compression of the cutaneous fibers from the third interspace which, at the distal carpal tunnel, are clumped superficially in the anteroulnar portion of the median nerve just beneath the transverse ligament. PMID- 8232395 TI - Focal myositis of the temporal muscle. AB - Focal myositis is a rare inflammatory disease of the skeletal muscle that may involve any part of the body. We present a 19-year-old man with a short history of a painful pseudotumor in the right temporal region. MRI pictures showed a swollen right temporal muscle and muscle biopsy revealed the characteristic histological and immunocytochemical findings of polymyositis. PMID- 8232396 TI - Predictive value of nerve conduction measurements at the carpal tunnel. AB - We compared the predictive values of three measurements of sensory conduction of the median nerve at the carpal tunnel (maximum latency difference [MLD], 8-cm latency [S8], and 14-cm latency [S14]) in 2334 hands of industrial workers, workers' compensation patients, and students. The MLD was determined by the centimetric technique. The MLD was the most sensitive and efficient measurement for predicting carpal tunnel syndrome (CTS). An MLD > or = 0.40 ms correctly identified 86.3% of 753 hands with CTS. The MLD correlated best with CTS and with the primary diagnostic category (CAT). MLD was the second factor selected in stepwise regression analysis for CAT (numbness was first and S8 was third). The MLD was the most important factor for predicting persistent or de novo CTS in a 5 year follow-up of 630 hands. Thus, the MLD was the most reliable nerve conduction study measurement for predicting current or future CTS in these subjects' hands. PMID- 8232397 TI - The influence of dose of microspheres on nerve blood flow, electrophysiology, and fiber degeneration of rat peripheral nerve. AB - Microsphere embolization of peripheral nerve results in a variable degree of ischemic fiber degeneration. To enhance the utility of the model, we evaluated the relationship between dose of microspheres to the supplying arteries of the sciatic-tibial nerve to nerve blood flow (NBF), electrophysiology, morphology, and behavioral changes. There was considerable variability in the effect of embolization on nerve pathology in individual nerves. However, the dose of microspheres regressed with the degree of hindlimb paresis, reduction in NBF, degree of fiber pathology, and ischemic conduction failure of the tibial nerve, evaluated at day 7. All nerves with severe (grade 4) fiber degeneration had flows of < 3 mL x 100 g-1 x min-1. We conclude that it is possible to predict with a high degree of accuracy the severity of fiber degeneration by the degree of NBF reduction and by the electrophysiologic abnormalities. PMID- 8232398 TI - Practice parameter for electrodiagnostic studies in carpal tunnel syndrome: summary statement. American Association of Electrodiagnostic Medicine, American Academy of Neurology, American Academy of Physical Medicine and Rehabilitation. AB - Carpal tunnel syndrome (CTS) is the most common peripheral mononeuropathy and often requires one or more therapies including surgery. The results of electrodiagnostic (EDX) studies have been found to be highly sensitive and specific for the diagnosis of CTS. This document defines the standards, guidelines, and options for EDX studies of CTS based on a critical review of the literature. PMID- 8232399 TI - Literature review of the usefulness of nerve conduction studies and electromyography for the evaluation of patients with carpal tunnel syndrome. AAEM Quality Assurance Committee. AB - The sensitivity and specificity of nerve conduction studies (NCS's) and electromyography (EMG) for the diagnosis of carpal tunnel syndrome (CTS) were evaluated by a critical review of the literature. With a search of the medical literature in English through May 1991, 165 articles were identified and reviewed on the basis of six criteria of scientific methodology. The findings of 11 articles that met all six criteria and the results of 48 additional studies that met four or five criteria are presented. We concluded that median sensory and motor NCS's are valid and reproducible clinical laboratory studies that confirm a clinical diagnosis of CTS with a high degree of sensitivity and specificity. Clinical practice recommendations are made based on a comparison of the sensitivities of the several different median nerve conduction study (NCS) techniques. PMID- 8232400 TI - Constructing reference ranges. PMID- 8232401 TI - Coexistence of amyotrophic lateral sclerosis and Werdnig-Hoffmann disease within a family. PMID- 8232402 TI - Multifocal conduction block in n-hexane neuropathy. PMID- 8232403 TI - Posterior antebrachial cutaneous neuropathy associated with lateral elbow pain. PMID- 8232404 TI - Sensitivity of various electrophysiologic studies for the diagnosis of carpal tunnel syndrome. PMID- 8232405 TI - Electrophysiologic evaluation of the paraneoplastic syndrome in early stages. PMID- 8232406 TI - Electrophysiology of Microsporidia myositis in an AIDS patient. PMID- 8232407 TI - Are eccentric contractions required to induce the skeletal muscle fiber disruption that occurs following unaccustomed activity? PMID- 8232408 TI - Differential susceptibility to repetitive potassium induced contractures in diaphragm versus extensor digitorum longus muscles of mdx mice. PMID- 8232409 TI - Molecular cloning of a rac family protein kinase and identification of a serine/threonine protein kinase gene family of Entamoeba histolytica. AB - Eleven Entamoeba histolytica protein-serine/threonine-kinase gene segments were identified using the polymerase chain reaction (PCR) and degenerate oligonucleotide primers to conserved amino acids in subdomains VI and VIII of the catalytic domain of protein-serine/threonine kinases. These ameba gene segments were homologous to myosin light chain kinases, protein kinase C, phosphorylase b kinase, and kinases that regulate glucose repression in yeast and cell growth in mammalian cells. One of these PCR products, which was homologous to the Dictyostelium discoideum protein kinase 2, was used to identify a full-length protein-serine/threonine-kinase gene (Eh rac1) from an E. histolytica genomic library. The open reading frame of Eh rac1 was 409 amino acids long (encoding a 47-kDa protein) and included an amino terminal segment containing 87 mostly charged and polar amino acids and a 322-amino acid carboxyl terminal segment containing the catalytic domain. The catalytic domain of Eh rac1 was homologous to the rac family of protein-serine/threonine-kinases, which are related to cAMP dependent protein kinases and protein kinase Cs. Southern blots of ameba DNA showed that the Eh rac1 gene was present as a single copy in all strains tested, however pathogenic amebae expressed four times more Eh rac1 mRNAs than did nonpathogenic amebae. These studies suggest that E. histolytica, a primitive unicellular eukaryote, has a complex protein kinase family. PMID- 8232410 TI - Reduced purine accumulation is encoded on an amplified DNA in Leishmania mexicana amazonensis resistant to toxic nucleosides. AB - Nucleoside analogs are potential anti-Leishmania agents. To better understand how these compounds might lose their effectiveness, Leishmania were independently selected for resistance to inosine dialdehyde or tubercidin. Each of the resistant cells exhibited resistance to inosine dialdehyde and tubercidin as well as to formycin B and allopurinol ribonucleoside. Resistant cells had a greatly reduced capability of accumulating exogenous adenosine, guanosine, thymidine and guanine. This decreased ability to accumulate nucleosides and at least one nucleobase appeared to be due to reduced activity of a number of distinct purine transporters, as the differences between purine metabolizing enzymes were not sufficiently different to account for the decreased accumulation capability. The resistance to toxic nucleosides and the decreased ability to accumulate purines were due to the presence in the resistant cells of an extrachromosomal DNA approximately 55 kb in size. The extrachromosomal DNA was not detected in wild type cells or revertants which have lost resistance to toxic nucleosides. Except for a 1.2-kb difference, the extrachromosomal DNA from both independently selected resistant cells appeared to be identical. The resistant cells contained 2-4 times as much DNA homologous to the extrachromosomal DNA as compared to wild type cells. When cloned into an E. coli/Leishmania shuttle vector, a portion of the amplified DNA had the ability to confer upon wild-type cells resistance to the toxic purine nucleoside analogs tubercidin and inosine dialdehyde. These transformed cells also exhibited a decreased ability to accumulate non-toxic purine nucleosides. PMID- 8232411 TI - The hypoxanthine-guanine-xanthine phosphoribosyltransferase from Tritrichomonas foetus has unique properties. AB - Tritrichomonas foetus, an anaerobic, flagellated protozoan parasite, is incapable of de novo purine nucleotide synthesis, and depends primarily on the salvage of purine bases from the host. The hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase) from this organism has been purified to homogeneity by ammonium sulfate precipitation and Sephacryl-HR100 gel filtration, followed by anion exchange FPLC. Hypoxanthine, guanine and xanthine phosphoribosyltransferase activities co-eluted in all the purification steps, suggesting that they are associated with the same enzyme protein. The molecular mass of the native protein, as estimated by gel filtration, is 24 kDa. The molecular mass estimated from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is also 24 kDa. Non-denaturing polyacrylamide gel electrophoresis of the purified protein, followed by activity staining with either [14C]hypoxanthine, [14C]guanine or [14C]xanthine, also demonstrates that the enzyme is a monomer of 24 kDa. This monomeric structure is distinctive from all the other reported PRTases which are either dimers or tetramers. Furthermore, unlike the mammalian HGPRTase, which is heat stable, the T. foetus enzyme is heat labile. Kinetic studies with the purified T. foetus HGXPRTase showed that the apparent Kms for hypoxanthine, guanine and xanthine were 4.1 microM, 3.8 microM and 52.4 microM respectively. This recognition of xanthine as a substrate by the parasite enzyme with only about a 10-fold higher Km value than those for hypoxanthine and guanine distinguishes it from the mammalian HGPRTase, which cannot use xanthine as a substrate, as well as the HGXPRTases of Eimeria tenella and Plasmodium falciparum, which are dimers, with xanthine about 100-times less proficient as a substrate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232412 TI - Cloning and functional analysis of an extrachromosomally amplified multidrug resistance-like gene in Leishmania enriettii. AB - The goal of this work was to investigate the mechanism of drug resistance in Leishmania enriettii as a model system for drug resistance both in human leishmaniasis and on other parasitic diseases. Parasites were selected in increasing concentrations of vinblastine, an inhibitor of microtubule assembly, and resistant clones were isolated which grew in concentrations 5-30 times the IC50 (30 micrograms ml-1) of parental cells. The vinblastine-resistant parasites were also resistant to puromycin, an unrelated drug which inhibits protein synthesis. This cross-resistance to unrelated drugs had previously been observed in mammalian cells and recently in L. donovani. The proposed mechanism for this cross-resistance is drug efflux mediated by increased expression of a P glycoprotein molecule encoded by a multidrug resistance (mdr) gene. Here we report the identification, cloning and sequencing of an mdr-like gene from L. enriettii, lemdr1, and demonstrate that this gene is amplified on an extrachromosomal circle of 35-40 kb in vinblastine-resistant L. enriettii. The longest open reading frame in the cloned gene is 1280 amino acids with a predicted protein of 140 kDa. The predicted protein has a structure similar to that for all other reported P-glycoproteins namely 12 transmembrane domains and 2 ATP binding sites, arranged in 2 similar half-molecules. Comparison of the primary amino acid sequence with other known mdr gene products demonstrates a significant homology with 37% amino acid identity with human mdr1 and 83% identity with the L. donovani ldmdr1 gene. The lemdr1 gene was cloned in the expression vector pALTNEO and transfected into wild-type L. enriettii and the resulting transfected cells were resistant to vinblastine but at lower levels than in the selected mutant cells. PMID- 8232413 TI - Molecular cloning and characterization of actin genes from Echinococcus granulosus. AB - An Echinococcus granulosus genomic library has been screened with a mouse beta actin cDNA probe. Two clones carrying DNA fragments of about 15 kb, possibly derived from the same genome region, have been isolated. This 15-kb genomic region includes 2 actin-related sequences (EgactI and EgactII) separated by about 4 kb. The nucleotide sequences of both genes were determined. The EgactI sequence presents no introns, but an intron of 591 bp was observed in the EgactII sequence. The genes potentially encode 375 and 376 amino-acid-long actins, respectively, with a homology of 85.3%. The deduced amino acid sequences from both genes were compared to the actin sequences from other organisms, showing similarities ranging from 63.5% to 90.6%. The nucleotide sequence of a partial actin cDNA clone has been determined. The deduced amino acids sequence showed a homology of 90.3% and 88.0% in relation to the EgactI and EgactII sequences respectively, suggesting the existence of at least one more actin gene in E. granulosus. This hypothesis is reinforced by the number of bands detected in the Southern blot analysis. Experiments based on the amplification of DNA segments using 3'-specific actin primers indicate that the EgactI gene is transcribed in protoscoleces. PMID- 8232414 TI - N,N'-thiophene-substituted polyamine analogs inhibit mammalian host cell invasion and intracellular multiplication of Trypanosoma cruzi. AB - We studied the effects of two N,N'-thiophene-substituted polyamine analogs (MDL 28302 and MDL 29431) on the capacities of Trypanosoma cruzi, the etiologic agent of Chagas' disease, to invade and multiply within a mammalian host cell. Both compounds inhibited infectivity significantly in a time- and concentration dependent manner. This inhibition resulted from a selective effect on the parasite, because pretreatment of T. cruzi but not host cell cultures with either MDL 28302 or MDL 29431 reduced infectivity. The parasite gradually recovered its infective capacity after removal of unincorporated polyamine analog, denoting the reversible nature of the inhibitory effect. Some biochemical modification of MDL 28302 and MDL 29431 appeared to be required for their inhibitory activities to be exerted, since the effects of these drugs on T. cruzi infectivity were abrogated by MDL 72527, a drug known to inhibit polyamine oxidase (PAO) activity specifically. Supporting the notion of that products of MDL 28302 and MDL 29431 oxidation by PAO were involved in the activity of these compounds was the finding that PAO competitive substrates (N1-acetylspermine and N1-acetylspermidine) also abolished the inhibition of T. cruzi infectivity mediated by MDL 28302 or MDL 29431. However, we can not rule out that MDL 72527 and the PAO competitive substrates might have altered an alternative mechanism because no significant polyamine oxidase activity could be demonstrated in preparations of lysed or intact T. cruzi in assays monitoring conversion of [14C]spermine to [14C]spermidine. When either MDL 28302 or MDL 29431 was added to infected cell cultures, a marked reduction in the rate of intracellular parasite growth ensued.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232415 TI - The virulent Saimiri-adapted Palo Alto strain of Plasmodium falciparum does not express the ring-infected erythrocyte surface antigen. AB - The Palo Alto strain of Plasmodium falciparum is highly virulent for the Saimiri sciureus monkey. We have observed that these parasites do not express the Ring infected erythrocyte surface antigene (RESA) gene. Immunoblots indicated that the Pf155/RESA protein was absent. The RESA mRNA could not be detected. Polymerase chain reaction and Southern blot analysis demonstrated that this lack of expression is due to gene rearrangements. The majority of the Palo Alto parasites have a deletion of the entire RESA gene, whereas in a minor fraction the RESA sequences remain detectable, but the 5' miniexon 1 is inverted. These data show that the RESA protein is dispensable for in vivo parasite growth, at least in Saimiri monkeys. PMID- 8232416 TI - Galactofuranose-containing glycoconjugates of epimastigote and trypomastigote forms of Trypanosoma cruzi. AB - Antiserum to LPPG, a lipopeptidophosphoglycan originally described on the surface of Trypanosoma cruzi epimastigotes of the Y strain, and antibodies to furanoic galactose (galf) were obtained in rabbits. A micromethod for the extraction and purification of LPPG from a limited amount of parasites is described. Analysis by Western blots of the purified glycoconjugate probed with both antisera confirmed the presence of galf-containing LPPG-like molecules in 10 different strains and clones of T. cruzi. An analogous approach indicated that trypomastigotes also contain LPPG-like components. Quantitation experiments allowed to calculate an average value of 1.0 x 10(7) LPPG molecules per epimastigote cell and 0.16 x 10(7) LPPG-like molecules per trypomastigote cell. Immunoelectron microscopy has shown a homogenous distribution of LPPG on the surface of epimastigotes. The trypomastigote population, however, is highly heterogenous with no more than 15% of the parasites being labeled by the anti-LPPG serum. Intense labeling has also been found in vesicles inside the epimastigote and trypomastigote forms. The distribution of galf epitopes among glycoconjugates of epimastigotes and trypomastigotes was further investigated. It was shown that galf units in epimastigotes are bound to low molecular mass compounds which co-migrate with LPPG whereas in trypomastigotes they have been found in both low molecular mass LPPG-like molecules and glycoproteins of 80-90 kDa. Direct chemical evidence for the presence of galf residues in the N-linked oligosaccharide chains of these surface glycoproteins has been obtained. Finally, the natural antigenicity of LPPG and galf in chronic Chagas' disease was investigated. It was found that all chronic chagasic sera investigated recognize this glycoconjugate and that an important part of such recognition can be attributed to galf residues. Furthermore, no correlation among reactivity to LPPG, strain zymodeme and clinical forms of the disease was found. PMID- 8232417 TI - Characterization of a divergent glycosomal microbody phosphoglycerate kinase from Trypanosoma brucei. AB - There are 3 loci in the phosphoglycerate kinase (PGK) gene complex of Trypanosoma brucei. The PGK-A gene product, which we term 56PGK, is targeted to glycosomal microbodies and is highly homologous to the parasite's 2 known PGKs (one cytoplasmic and one glycosomal). However, 56PGK contains an 80 amino acid insertion as well as numerous substitutions compared to the other PGKs. The complementation and kinetic analyses described here demonstrate that 56PGK is an authentic phosphoglycerate kinase--the largest yet described. When expressed in Escherichia coli, 56PGK complements the pgk- phenotype. 56PGK was expressed as a fusion protein and purified to near homogeneity. The Michaelis constants are similar to those of other PGKs, being 0.12 and 2.4 mM for Mg-ATP and 3 phosphoglycerate, respectively. As with other T. brucei PGKs, ATP but not GTP or ITP can serve as a phosphate donor during catalysis. No evidence was obtained for phosphate transfer to atypical substrates. 56PGK shows sulfate inhibition at all concentrations tested, rather than the sulfate activation observed with yeast PGK. PMID- 8232418 TI - Repetitive sequences in the ribosomal intergenic spacer of Trypanosoma cruzi. AB - A fragment of Trypanosoma cruzi ribosomal intergenic spacer (IGS) located at 6.7 kb from the 3' end of the 24S rRNA gene was analyzed. This IGS fragment is characterized by the presence of three types of repetitive elements (designated Spacer Repetitive Elements, SRE), short direct repeats (5-6 bp) and chi-like recombinational sequences. SRE elements are composed of relatively short repeats (43-145 bp) which show variabilities consisting of nucleotide changes, insertions and deletions. SRE-1 element (145 bp) has a short oligo(dA) tail at the end of the repeat and can be found flanked by other SRE elements. SRE elements are species-specific, suggesting that probes based on them may be diagnostic for Trypanosoma cruzi. PMID- 8232419 TI - Using simultaneous, tandem gene replacements to study expression of the multicopy ubiquitin-fusion (FUS) gene family of Trypanosoma cruzi. AB - Many genes in trypanosomes exist as members of multicopy gene families. Due to this fact it is frequently difficult to determine if specific members of a gene family are expressed. We describe here a strategy for simultaneous tandem gene replacement in T. cruzi which leads to the replacement of the gene of interest by a silent reporter gene, the expression of which can be assayed in stable transformants. To determine if the FUS1 gene (one of 5 copies of the ubiquitin fusion, FUS, gene family) was expressed, stable G418-resistant transformants were isolated in which the tandemly arrayed CUB2.65 and FUS1 genes were precisely replaced by the neomycin phosphotransferase (neo(r)) and chloramphenicol acetyltransferase (CAT) genes, respectively. All stable clones carrying the tandem gene replacements were shown to express the CAT activity indicating that FUS1 is expressed in mid-log epimastigotes. Northern blot analysis of parasites carrying the tandem gene replacements indicated that at least one other member of the FUS gene family is expressed and that there were no apparent polar effects on the expression of genes downstream of the replacement events. These experiments have demonstrated the utility of tandem gene replacements as a means of inserting a nonselected reporter gene into the chromosome, facilitating the molecular genetic analysis of the expression of multicopy gene families. PMID- 8232420 TI - Studies on mitochondrial ATPase of Leishmania donovani using digitonin permeabilized promastigotes. AB - Mitochondrial ATPase of Leishmania donovani was characterized using digitonin permeabilized promastigotes and the results were compared with those from isolated mitochondria. Maximum mitochondrial ATPase activity was obtained in promastigotes permeabilized with digitonin at a final concentration of 20 microM and the specific activity of the enzyme was 46% and 57% higher than that of homogenized and sonicated promastigotes, respectively. At concentrations above 20 microM digitonin inhibited ATPase activity and the degree of inhibition increased with increasing concentrations of the detergent. The ATPase activity of promastigotes remained DCCD-sensitive when permeabilized with digitonin at concentrations up to 120 microM but the enzyme became increasingly resistant to this inhibitor as digitonin concentrations were increased to 140 microM and more, indicating the loss of functional activity of the enzyme. The pH and temperature optima for mitochondrial ATPase were determined to be 7.5 and 30 degrees C, respectively. Mg2+ ions were essential for ATPase activity but free Mg2+ ions were found to be inhibitory. A Mg2+/ATP ratio of 1:3 supported the optimum ATPase activity. Sulfite and hexanol activated the enzyme but failed to prevent the inhibition by free Mg2+ ions. The results indicate that digitonin-permeabilized promastigotes provide an ideal system for studying the mitochondrial ATPase of L. donovani. PMID- 8232421 TI - Uptake and turnover of glucose in Leishmania donovani. AB - Glucose uptake and metabolism by Leishmania donovani promastigotes was studied using D-[14C]glucose in combination with the silicone oil centrifugation technique on organisms preadapted to different growth rates and glucose availability in the chemostat. The uptake step was differentiated from the subsequent metabolism by separation in time rather than by using non metabolisable analogues. The uptake of glucose was measured as a function of time and/or the external glucose concentration on cells grown at high or low growth rate with glucose either as growth rate-limiting substrate, or present in excess. Glucose uptake as a function of its external concentration could be described as consisting of two components (1) a rapid equilibration owing to facilitated diffusion, followed by (2) a much slower uptake that involves an enzymatic component. This slower accumulation of label could be explained as the conversion of glucose into metabolites and a storage carbohydrate. Uptake experiments in the presence of inhibitors indicated that the conversion of glucose was an energy dependent process. These experiments indicate that the active uptake of glucose by L. donovani, as reported by others does not occur across the plasma membrane and should be reinterpreted as the intracellular conversion of glucose into metabolites and storage carbohydrate. PMID- 8232422 TI - The cDNA sequence of an abundant Entamoeba histolytica 20-kilodalton protein containing four repetitive domains. PMID- 8232423 TI - An Eimeria tenella gene encoding a protein with homology to the nucleotide transhydrogenases of Escherichia coli and bovine mitochondria. PMID- 8232424 TI - Conservation of gRNA gene cassette structure in African trypanosomes despite divergence in the defining flanking repeats. PMID- 8232425 TI - Sequences of three Trypanosoma congolense maxicircle genes allow prediction of regions encoding transcripts that undergo extensive RNA editing. PMID- 8232426 TI - cDNA cloning and recombinant expression of collagen-binding and complement inhibitor activity of Taenia solium paramyosin (AgB). PMID- 8232427 TI - The dihydroorotate dehydrogenase gene homologue of Plasmodium falciparum. PMID- 8232428 TI - Neonatal complications after the administration of indomethacin for preterm labor. AB - BACKGROUND: The use of indomethacin as a tocolytic agent in pregnant women appears to be accompanied by a low incidence of neonatal complications. However, the neonatal effects of indomethacin have been studied primarily in infants born after 32 weeks' gestation. This study was designed to examine the incidence of neonatal complications in very premature infants. METHODS: We identified 57 infants delivered at or before 30 weeks' gestation whose mothers had been treated with indomethacin for preterm labor and matched them with 57 infants whose mothers had not received indomethacin. The infants in the two groups were matched for sex, gestational age at delivery (mean [+/- SD], 27.6 +/- 2.0 weeks), exposure to betamethasone for 24 hours or more before delivery, and rupture of membranes 24 hours or more before delivery. RESULTS: There were no significant differences between the two groups in birth weight, Apgar scores, cord-blood gas values, frequency of multiple gestation, or incidence of respiratory distress syndrome. The proportion of infants who required exogenous surfactant was similar, as were ventilator settings at 24 hours, the incidence of chronic lung disease, and the incidence of sepsis. The infants exposed to indomethacin had a lower urine output and higher serum creatinine concentrations during the first three days after delivery. More indomethacin-exposed infants had necrotizing enterocolitis (29 percent vs. 8 percent, P = 0.005), intracranial hemorrhage grade II to IV (28 percent vs. 9 percent, P = 0.02), and patent ductus arteriosus (62 percent, vs. 44 percent, P = 0.05). More indomethacin-exposed infants with a patent ductus arteriosus required surgical ligation because of either a lack of initial response or a reopening of the duct after postnatal indomethacin therapy (50 percent vs. 20 percent of the unexposed infants, P = 0.05). CONCLUSIONS: Antenatal indomethacin therapy for preterm labor appears to increase the risk of serious neonatal complications in infants born at or before 30 weeks' gestation. PMID- 8232429 TI - Recombinant interferon alfa-2b combined with a regimen containing doxorubicin in patients with advanced follicular lymphoma. Groupe d'Etude des Lymphomes de l'Adulte. AB - BACKGROUND: Interferon alfa and cytotoxic drugs have synergistic effects in patients with non-Hodgkin's lymphoma. In 1986, we designed a clinical trial to evaluate the benefit of concomitant administration of recombinant interferon alfa with a regimen containing doxorubicin in patients with follicular non-Hodgkin's lymphoma. METHODS: The trial involved 242 patients with advanced low-grade follicular non-Hodgkin's lymphoma selected on the basis of clinical, radiographic, and biologic criteria. All patients were treated with a regimen consisting of cyclophosphamide, doxorubicin, teniposide, and prednisone (CHVP), given monthly for six cycles and then every two months for one year. After randomization, 123 patients also received interferon alfa-2b at a dosage of 5 million units three times weekly for 18 months. The remaining 119 patients received chemotherapy alone. RESULTS: As compared with the patients treated with CHVP only, the patients treated with CHVP plus interferon alfa had a higher overall rate of response (85 percent vs. 69 percent, P = 0.006), a longer median event-free survival (34 months vs. 19 months, P < 0.001), and a higher rate of survival at 3 years (86 percent vs. 69 percent, P = 0.02). Granulocyte toxicity was greater in the patients treated with CHVP plus interferon alfa than in those treated with CHVP alone. There were no treatment-related deaths. Interferon alfa had to be stopped because of toxic effects (fatigue and hepatitis) in 13 patients (11 percent). CONCLUSIONS: The addition of interferon alfa to a regimen containing doxorubicin increased the rate of response, event-free survival, and overall survival in patients with advanced follicular non-Hodgkin's lymphoma, without serious toxicity, although some patients were unable to tolerate the side effects. PMID- 8232430 TI - The effects of tissue plasminogen activator, streptokinase, or both on coronary artery patency, ventricular function, and survival after acute myocardial infarction. AB - BACKGROUND: Although it is known that thrombolytic therapy improves survival after acute myocardial infarction, it has been debated whether the speed with which coronary-artery patency is restored after the initiation of therapy further affects outcome. METHODS: To study this question, we randomly assigned 2431 patients to one of four treatment strategies for reperfusion: streptokinase with subcutaneous heparin; streptokinase with intravenous heparin; accelerated-dose tissue plasminogen activator (t-PA) with intravenous heparin; or a combination of both activators plus intravenous heparin. Patients were also randomly assigned to cardiac angiography at one of four times after the initiation of thrombolytic therapy: 90 minutes, 180 minutes, 24 hours, or 5 to 7 days. The group that underwent angiography at 90 minutes underwent it again after 5 to 7 days. RESULTS: The rate of patency of the infarct-related artery at 90 minutes was highest in the group given accelerated-dose t-PA and heparin (81 percent), as compared with the group given streptokinase and subcutaneous heparin (54 percent, P < 0.001), the group given streptokinase and intravenous heparin (60 percent, P < 0.001), and the group given combination therapy (73 percent, P = 0.032). Flow through the infarct-related artery at 90 minutes was normal in 54 percent of the group given t-PA and heparin but in less than 40 percent in the three other groups (P < 0.001). By 180 minutes, the patency rates were the same in the four treatment groups. Reocclusion was infrequent and was similar in all four groups (range, 4.9 to 6.4 percent). Measures of left ventricular function paralleled the rate of patency at 90 minutes; ventricular function was best in the group given t PA with heparin and in patients with normal flow through the infarct-related artery irrespective of treatment group. Mortality at 30 days was lowest (4.4 percent) among patients with normal coronary flow at 90 minutes and highest (8.9 percent) among patients with no flow (P = 0.009). CONCLUSIONS: This study supports the hypothesis that more rapid and complete restoration of coronary flow through the infarct-related artery results in improved ventricular performance and lower mortality among patients with myocardial infarction. This would appear to be the mechanism by which accelerated t-PA therapy produced the most favorable outcome in the GUSTO trial. PMID- 8232431 TI - Brief report: correction of X-linked lymphoproliferative disease by transplantation of cord-blood stem cells. PMID- 8232432 TI - Images in clinical medicine. Skin and nail lesions in endocarditis. PMID- 8232433 TI - Rabies. PMID- 8232434 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 47-1993. Presentation of case. A 28-year-old man with recurrent ventricular tachycardia and dysfunction of multiple organs. PMID- 8232435 TI - Outcomes of extremely immature infants--a perinatal dilemma. PMID- 8232436 TI - The open-artery theory is alive and well--again. PMID- 8232437 TI - Rethinking professional courtesy. PMID- 8232438 TI - Hospital administrative costs. PMID- 8232439 TI - Hospital administrative costs. PMID- 8232440 TI - Hospital administrative costs. PMID- 8232441 TI - Hospital administrative costs. PMID- 8232442 TI - Hospital administrative costs. PMID- 8232443 TI - Hospital administrative costs. PMID- 8232444 TI - Administrative costs and health care reform. PMID- 8232445 TI - Administrative costs and health care reform. PMID- 8232446 TI - Mucosa-associated lymphoid-tissue lymphoma. PMID- 8232447 TI - Reversible lymphomas. PMID- 8232448 TI - Group B streptococcal disease in adults. PMID- 8232449 TI - Group B streptococcal disease in adults. PMID- 8232450 TI - Aerosolized tobramycin in patients with cystic fibrosis. PMID- 8232451 TI - Prenatal prediction of risk of the fetal hydantoin syndrome. PMID- 8232452 TI - Causes of death in injection-drug users. PMID- 8232453 TI - Inadvertent placement of an advertisement. PMID- 8232454 TI - The framework of health care reform. PMID- 8232455 TI - Changes in the delivery of care under comprehensive health care reform. PMID- 8232456 TI - Triggering of acute myocardial infarction by heavy physical exertion. Protection against triggering by regular exertion. Determinants of Myocardial Infarction Onset Study Investigators. AB - BACKGROUND: Despite anecdotal evidence suggesting that heavy physical exertion can trigger the onset of acute myocardial infarction, there have been no controlled studies of the risk of myocardial infarction during and after heavy exertion, the length of time between heavy exertion and the onset of symptoms (induction time), and whether the risk can be modified by regular physical exertion. To address these questions, we collected data from patients with confirmed myocardial infarction on their activities one hour before the onset of myocardial infarction and during control periods. METHODS: Interviews with 1228 patients conducted an average of four days after myocardial infarction provided data on their usual annual frequency of physical activity and the time, type, and intensity of physical exertion in the 26 hours before the onset of myocardial infarction. We compared the observed frequency of heavy exertion (6 or more metabolic equivalents) with the expected values using two types of self-matched analyses based on a new case-crossover study design. The low frequency of heavy exertion during the control periods was validated by data from a population-based control group of 218 subjects. RESULTS: Of the patients, 4.4 percent reported heavy exertion within one hour before the onset of myocardial infarction. The estimated relative risk of myocardial infarction in the hour after heavy physical exertion, as compared with less strenuous physical exertion or none, was 5.9 (95 percent confidence interval, 4.6 to 7.7), Among people who usually exercised less than one, one to two, three to four, or five or more times per week, the respective relative risks were 107 (95 percent confidence interval, 67 to 171), 19.4 (9.9 to 38.1), 8.6 (3.6 to 20.5), and 2.4 (1.5 to 3.7). Thus, increasing levels of habitual physical activity were associated with progressively lower relative risks. The induction time from heavy exertion to the onset of myocardial infarction was less than one hour, and symptoms usually began during the activity. CONCLUSIONS: Heavy physical exertion can trigger the onset of acute myocardial infarction, particularly in people who are habitually sedentary. Improved understanding of the mechanisms by which heavy physical exertion triggers the onset of myocardial infarction and the manner in which regular exertion protects against it would facilitate the design of new preventive approaches. PMID- 8232457 TI - Physical exertion as a trigger of acute myocardial infarction. Triggers and Mechanisms of Myocardial Infarction Study Group. AB - BACKGROUND: It is controversial whether the onset of myocardial infarction occurs randomly or is precipitated by identifiable stimuli. Previous studies have suggested a higher risk of cardiac events in association with exertion. METHODS: Consecutive patients with acute myocardial infarction were identified by recording all admissions to our hospital in Berlin and by monitoring a general population of 330,000 residents in Augsburg, Germany. Information on the circumstances of each infarction was obtained by means of standardized interviews. The data analysis included a comparison of patients with matched controls and a case-crossover comparison (one in which each patient serves as his or her own control) of the patient's usual frequency of exertion with the last episode of exertion before the onset of myocardial infarction. RESULTS: From January 1989 through December 1991, 1194 patients (74 percent of whom were men; mean age [+/- SD], 61 +/- 9 years) completed the interview 13 +/- 6 days after infarction. We found that 7.1 percent of the case patients had engaged in physical exertion (> or = 6 metabolic equivalents) at the onset of infarction, as compared with 3.9 percent of the controls at the onset of the control event. For the patients as compared with the matched controls, the adjusted relative risk of having engaged in strenuous physical activity at the onset of infarction or the control event was 2.1 (95 percent confidence interval, 1.1 to 3.6). The case crossover comparison yielded a similar relative risk of 2.1 (95 percent confidence interval, 1.6 to 3.1) for having engaged in strenuous physical activity within one hour before myocardial infarction. Patients whose frequency of regular exercise was less than four and four or more times per week had relative risks of 6.9 and 1.3, respectively (P < 0.01). CONCLUSIONS: A period of strenuous physical activity is associated with a temporary increase in the risk of having a myocardial infarction, particularly among patients who exercise infrequently. These findings should aid in the identification of the triggering mechanisms for myocardial infarction and improve prevention of this common and serious disorder. PMID- 8232458 TI - A prospective study of diarrhea and HIV-1 infection among 429 Zairian infants. AB - BACKGROUND: Persistent diarrhea is a prominent feature of the acquired immunodeficiency syndrome in adults, but its cause and its effect on children with human immunodeficiency virus (HIV) infection are largely unknown, particularly in Africa. METHODS: We studied a birth cohort of 429 infants born to HIV-positive or HIV-negative mothers in Zaire to determine the incidence of acute, recurrent (> or = 2 episodes), and persistent (> or = 14 days) diarrhea; outcome; and risk factors. RESULTS: Of the 238 infants whose mothers were HIV positive, 53 were infected, 139 were uninfected, and the HIV status of 46 could not be determined. As compared with uninfected infants, infected infants had higher incidence rates for acute diarrhea (170 vs. 100 episodes per 100 child years, P = 0.003), recurrent diarrhea (21 vs. 11, P = 0.12), and persistent diarrhea (19 vs. 4, P < 0.003). Persistent diarrhea developed in 11 HIV-infected infants; all but 1 died. It also developed in 19 uninfected infants; all but 1 survived. The prevalence of stool pathogens was similar in the two groups. In a multivariate model, persistent diarrhea in an infant was independently associated with symptomatic HIV type 1 infection in the mother (relative hazard, 1.5; P = 0.08). The incidence of persistent diarrhea in the uninfected infants of seropositive mothers was nearly double that in the uninfected infants of seronegative mothers (4.9 vs. 2.7 episodes per 100 child-years), and the risk increased if the mother died (relative hazard, 10.4). Significant growth impairment and severe immunosuppression occurred in the six to eight weeks before the onset of persistent diarrhea. CONCLUSIONS: In Zaire, infants with HIV infection have an 11-fold increased risk of death from diarrhea, largely persistent diarrhea, which is often preceded by recurrent episodes of acute diarrhea, malnutrition, or immunosuppression. Illness and death of the mother increase that risk, even among her uninfected infants. PMID- 8232459 TI - Growth and pituitary-adrenal function in children with severe asthma treated with inhaled budesonide. AB - BACKGROUND: The increased use of inhaled corticosteroids in the management of asthma raises concern about the safety of these drugs in children. We sought to determine the safety of long-term administration of inhaled budesonide in young children with asthma. METHODS: We studied 15 children 2 to 7 years old who had severe perennial asthma. They inhaled 100 micrograms of budesonide twice daily for three to five years. Efficacy was assessed by serial evaluation of respiratory symptoms and the need for other medications, and safety by serial evaluation of height, height velocity, weight, bone age, and pituitary-adrenal function. RESULTS: The severity of asthma decreased within the first month after the initiation of therapy, as demonstrated by a 58 percent reduction in the number of days with symptoms of asthma and a 75 percent decrease in the use of bronchodilators. This improvement was maintained thereafter. The growth pattern of all patients, including their height, weight, and bone age, was normal (as compared with standard normal values) throughout the treatment period. Pituitary adrenal function was not adversely affected by the treatment, as demonstrated by normal serum cortisol concentrations in the morning and 60 minutes after stimulation with corticotropin, normal 24-hour serum cortisol concentrations (mean [+/- SD] of samples collected at 30-minute intervals for 24 hours, 8.4 +/- 4.2 micrograms per deciliter [232 +/- 116 nmol per liter]), and normal urinary cortisol excretion (34 +/- 9 micrograms [95 +/- 25 nmol] per day). CONCLUSIONS: Prolonged administration of 200 micrograms of inhaled budesonide daily to young children with severe asthma does not impair growth or pituitary-adrenal function. PMID- 8232460 TI - Images in clinical medicine. Polyp of the appendix. PMID- 8232461 TI - Clinical problem-solving. When going for the gold is not an option. PMID- 8232462 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 48-1993. A 27-year-old woman with mediastinal lymphadenopathy and relentless cor pulmonale. PMID- 8232463 TI - Is exercise beneficial--or hazardous--to your heart? PMID- 8232464 TI - Differences between inhaled and oral glucocorticoid therapy. PMID- 8232465 TI - Reform of the health care system and professional liability. PMID- 8232466 TI - Praise reform and start the litigation? PMID- 8232467 TI - Sudden death in young athletes. PMID- 8232468 TI - Sudden death in young athletes. PMID- 8232469 TI - Sudden death in young athletes. PMID- 8232470 TI - Sudden cardiac death during mountain hiking and downhill skiing. PMID- 8232471 TI - Endothelial dysfunction in microvascular angina. PMID- 8232472 TI - Endothelial dysfunction in microvascular angina. PMID- 8232473 TI - Endothelial dysfunction in microvascular angina. PMID- 8232474 TI - Endothelial dysfunction in microvascular angina. PMID- 8232475 TI - Endothelin production in mitral stenosis. PMID- 8232476 TI - Iron deficiency in children. PMID- 8232477 TI - Iron deficiency in children. PMID- 8232478 TI - Iron deficiency in children. PMID- 8232479 TI - Bismuth in infants with watery diarrhea. PMID- 8232480 TI - Bismuth in infants with watery diarrhea. PMID- 8232481 TI - Criticism of clinical problem-solving. PMID- 8232482 TI - Hantavirus-associated acute respiratory failure. PMID- 8232483 TI - Letter from Bosnia. PMID- 8232484 TI - Preventing asthma epidemics due to soybeans by dust-control measures. AB - BACKGROUND: The inhalation of soybean dust released during the unloading of soybeans into a silo caused outbreaks of asthma in Barcelona, Spain. This study was designed to determine morbidity due to asthma and serum IgE reactivity before and after the installation of filters in the silo. METHODS: We measured soybean allergen concentrations in 136 samples of air collected for 9 months before and 24 months after the installation of filters. We compared the number of days on which there was an unexpected increase in emergency room visits for asthma, the number of days on which the number of emergency room visits for asthma in one four-hour period was so high that it was unlikely to be due to chance, and the mean daily number of emergency room and intensive care unit admissions for asthma for a total of 60 months before and after filter installation. Serum IgE antibodies against soybean allergens were measured in 38 patients before and after filter installation. RESULTS: The concentration of airborne soybean allergens on days when soybeans were unloaded decreased from 324 to 25 U per cubic meter after the installation of filters (P < 0.001). The number of days on which there was an unusually large number of visits to the hospital for asthma and the number of days on which asthma was epidemic both decreased significantly (both P < 0.001), from 29 to 6 and from 18 to 0, respectively. The mean daily number of emergency room and intensive care unit admissions for asthma on days when soybeans were unloaded decreased from 8.3 to 5.4 and from 0.26 to 0.01, respectively (both P < 0.001). The mean serum IgE antibody concentrations in the 38 patients studied decreased from 2 Pharmacia reference units per milliliter to 1 (P < 0.001). CONCLUSIONS: Installing filters on silos to prevent airborne dissemination of allergenic soybean dust eliminates outbreaks of asthma caused by inhalation of the dust, thus supporting the idea that the avoidance of allergens helps prevent asthma. PMID- 8232485 TI - The effect of corticosteroids for acute optic neuritis on the subsequent development of multiple sclerosis. The Optic Neuritis Study Group. AB - BACKGROUND: Optic neuritis is often the first clinical manifestation of multiple sclerosis, but little is known about the effect of corticosteroid treatment for optic neuritis on the subsequent risk of multiple sclerosis. METHODS: We conducted a multicenter study in which 389 patients with acute optic neuritis (and without known multiple sclerosis) were randomly assigned to receive intravenous methylprednisolone (250 mg every six hours) for 3 days followed by oral prednisone (1 mg per kilogram of body weight) for 11 days, oral prednisone (1 mg per kilogram) alone for 14 days, or placebo for 14 days. Neurologic status was assessed over a period of two to four years. The patients in the first group were hospitalized for three days; the others were treated as outpatients. RESULTS: Definite multiple sclerosis developed within the first two years in 7.5 percent of the intravenous-methyl-prednisolone group (134 patients), 14.7 percent of the oral-prednisone group (129 patients), and 16.7 percent of the placebo group (126 patients). The adjusted rate ratio for the development of definite multiple sclerosis within two years in the intravenous-methylprednisolone group was 0.34 (95 percent confidence interval, 0.16 to 0.74) as compared with the placebo group and 0.38 (95 percent confidence interval, 0.17 to 0.83) as compared with the oral-prednisone group. The beneficial effect of the intravenous-steroid regimen appeared to lessen after the first two years of follow-up. Signal abnormalities on magnetic resonance imaging (MRI) of the brain were a strong indication of risk for the development of definite multiple sclerosis (adjusted rate ratio in patients with three or more lesions, 5.53; 95 percent confidence interval, 2.41 to 12.66). The beneficial effect of treatment was most apparent in patients with abnormal MRI scans at entry. CONCLUSIONS: In patients with acute optic neuritis, treatment with a three-day course of high-dose intravenous methylprednisolone (followed by a short course of prednisone) reduces the rate of development of multiple sclerosis over a two-year period. PMID- 8232486 TI - Brief report: recurrent acyclovir-resistant genital herpes in an immunocompetent patient. PMID- 8232487 TI - Images in clinical medicine. Vulvar herpes simplex infection. PMID- 8232488 TI - The effect of cost-containment policies on rates of coronary revascularization in California. AB - BACKGROUND: Lower rates of use of resources have been reported for the treatment of hospitalized patients covered by Medicaid than for privately insured patients. Cost-containment policies may exacerbate such differences in the use of hospital resources. We studied patients with ischemic heart disease who received care at nonfederal hospitals in California in 1983 (the year a Medicaid cost-containment program was implemented), in 1985, or in 1988. Within this sample of patients, we compared the rates of coronary revascularization (coronary-artery bypass surgery or coronary angioplasty) among patients covered by Medicaid, patients with private insurance covering fee-for-service care, and patients enrolled in a health maintenance organization (HMO). METHODS: Logistic-regression models were used to determine adjusted odds ratios for the use of coronary revascularization procedures in patients with different types of insurance, with control for demographic, clinical, and hospital characteristics. The study samples were made up of 49,167 patients in 1983, 47,809 in 1985, and 44,631 in 1988. RESULTS: The frequency of revascularization increased in all three insurance groups from 1983 to 1988, but it did so much faster in the fee-for-service and HMO groups than in the Medicaid group. Patients with private fee-for-service insurance were 1.66 times as likely as Medicaid patients to undergo revascularization in 1983 (P < 0.01), 2.01 times as likely in 1985 (P < 0.01) and 2.33 times as likely in 1988 (P < 0.01). Patients enrolled in HMOs were 0.96 times as likely as Medicaid patients to undergo revascularization in 1983 (P < 0.05), 1.23 times as likely in 1985 (P < 0.01), and 1.53 times as likely in 1988 (P < 0.01). CONCLUSIONS: The frequency of coronary revascularization in California in 1983 was nearly twice as high for patients with private fee-for-service insurance as for patients enrolled in HMOs or for Medicaid recipients. The implementation that year of stringent cost-control measures by Medicaid may explain the slower increase in the frequency of revascularization over five year among Medicaid recipients as compared with patients in the fee-for-service and HMO groups. Different incentives in fee-for-service and HMO practice may explain the lower frequency of revascularization among patients enrolled in HMOs, although the rates of increase for these two groups were about the same from 1983 to 1988. PMID- 8232489 TI - Control of chemotherapy-induced emesis. AB - The development of antiemetic drugs has been one of the most rewarding areas of oncologic research, since therapeutic advances in this area can result in immediate improvement in the quality of life for patients undergoing chemotherapy. Antiemetic therapy has progressed dramatically during the past decade and a half. Fifteen years ago, patients receiving cisplatin for the first time had a median of 12 vomiting episodes within the first 24 hours, whereas now more than 50 percent of such patients have no vomiting episodes at all. Theoretical and clinical challenges remain, however, in the effort to control chemotherapy-induced emesis. The mechanisms of anticipatory vomiting and delayed vomiting are still not understood, and consistently effective therapeutic approaches to these problems have yet to be developed. PMID- 8232490 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 49-1993. A 21-year-old woman with lifelong progressive interstitial lung disease. PMID- 8232491 TI - Air pollution and mortality. PMID- 8232492 TI - Corticosteroids and optic neuritis. PMID- 8232493 TI - Transforming the size and composition of the physician work force to meet the demands of health care reform. PMID- 8232494 TI - The future of the academic medical center under health care reform. PMID- 8232495 TI - FDA policy on women in drug trials. PMID- 8232496 TI - Women's health. PMID- 8232497 TI - Women's health. PMID- 8232498 TI - Primary human herpesvirus 6 infection in an adult. PMID- 8232499 TI - Primary human herpesvirus 6 infection in an adult. PMID- 8232500 TI - Primary human herpesvirus 6 infection in an adult. PMID- 8232501 TI - Withdrawal of digoxin from patients with chronic heart failure treated with angiotensin-converting-enzyme inhibitors. PMID- 8232502 TI - Withdrawal of digoxin from patients with chronic heart failure treated with angiotensin-converting-enzyme inhibitors. PMID- 8232503 TI - Withdrawal of digoxin from patients with chronic heart failure treated with angiotensin-converting-enzyme inhibitors. PMID- 8232504 TI - Clinical problem-solving: one more hypothesis. PMID- 8232505 TI - Clinical problem-solving: one more hypothesis. PMID- 8232506 TI - Interferon alfa plus chemotherapy for non-Hodgkin's lymphoma: five-year follow up. PMID- 8232507 TI - Inhaled steroid use and glaucoma. PMID- 8232508 TI - More on central venous pressure in space. PMID- 8232509 TI - Phencyclidine receptor binding as a probe of NMDA receptor functioning: implications for drug abuse research. PMID- 8232510 TI - Sigma, PCP, and NMDA Receptor Systems. Proceedings of a meeting. Baltimore, Maryland, September 27-28, 1989. PMID- 8232511 TI - A role for sigma binding in the antipsychotic profile of BMY 14802? AB - BMY 14802 was identified as a potential antipsychotic drug in traditional model systems, and this identification was confirmed in modern behavioral and electrophysiological systems. The drug appears to be atypical as an antipsychotic in its lack of activity in models predictive of the potential to produce extrapyramidal side effects and tardive dyskinesia. Indeed, this suggestion is corroborated by clinical findings to date. The atypical profile of BMY 14802 extends to its neurochemical actions and appears to find its basis in regionally selective, indirect modulation of the dopamine system. Furthermore, BMY 14802 exhibits interactions with sigma binding sites in vitro and in vivo, a notion supported by data from neurophysiological, behavioral, and biochemical investigations. BMY 14802 also appears to be neuroprotective in some model systems and may have utility in the treatment of stroke (Boissard et al. 1991). BMY 14802 appears to interact with 5-HT1A receptors, but this interaction does not seem to contribute significantly to the potential antipsychotic actions of the drug. Moreover, the formation of active metabolites of BMY 14802 does not appear to occur in animals or humans to an extent of physiological or behavioral relevance. If clinically efficacious, BMY 14802 may treat the symptoms of schizophrenia by a mechanism novel for antipsychotic drugs: regionally selective, indirect modulation of dopaminergic systems by specific interaction at sigma sites. PMID- 8232512 TI - Molecular biology of PCP and NMDA receptors. AB - The studies described demonstrate that rat brain mRNA directs the synthesis of at least four types of functional EAA receptors in the Xenopus oocyte system, whereas in this system NCB-20 cell mRNA directs the synthesis of only the NMDA type of EAA receptor. The NMDA channel expressed in the oocyte, using either rat brain mRNA or NCB-20 cell mRNA, exhibits the pharmacologic properties of the neuronal receptor, including the functional association with the PCP receptor located within the NMDA-gated channel. The demonstration that mRNA isolated from NCB-20 cells lacking functional NMDA-activated channels, but bearing PCP binding sites, can encode functional NMDA-activated channels in the oocyte indicates some defect or regulating step in posttranslational processing or insertion of the receptors into the plasma membrane in the cell of origin. This is the only cell line known to (1) have PCP receptors that appear to be associated with NMDA receptors and (2) provide a homogeneous, self-replicating population of cells that can be manipulated genetically and by changing the extracellular environment. Consequently, the NCB-20 cell line will be useful for the study of the NMDA receptor and its expression. PMID- 8232513 TI - Excitatory amino acid neurotoxicity in the developing brain. AB - EAA neurotransmitters participate in a variety of physiologic processes during central nervous system development. EAAs function as neurotransmitters but also regulate development of neuronal cytoarchitecture and neuronal connectivity. EAAs play a major role in several forms of activity-dependent synaptic plasticity including learning and memory, and stabilization and elimination of synaptic connections during development. Similar molecular mechanisms may underlie plastic changes during development and neuronal destruction by overaction of EAA receptors. A critical level of EAA neurotransmitter activity is required for normal development. However, there appears to be a continuum of physiologic reactions to EAA receptor activation: underactivation can retard or disrupt normal development, whereas overactivation can lead to neuronal injury and destruction (figure 8). The susceptibility of the brain to excitotoxicity is dramatically altered during postnatal development. As a result of these changes, the contribution of NMDA receptor activation in excitotoxic brain injury may be greater during early periods of postnatal development, whereas non-NMDA receptors may make a greater contribution to excitotoxic injury in the adult brain. PMID- 8232514 TI - Isolation and characterization of an endogenous ligand for the PCP and sigma receptors from porcine, rat, and human tissue. PMID- 8232515 TI - Pharmacologic characterizations of sigma receptors. PMID- 8232516 TI - Studies of sigma receptors and metabolic responses to sigma ligands in the brain. PMID- 8232517 TI - Sigma receptors and signal transduction: negative modulation of signaling through phosphoinositide-linked receptor systems. PMID- 8232518 TI - Sigma and phencyclidine receptors in the brain-endocrine-immune axis. AB - We have demonstrated the absence of PCP receptors and the presence of sigma receptors in PBLs and rat pituitary, adrenal, testis, and ovary that have kinetic and pharmacologic characteristics comparable with the sigma receptor found in the CNS. The physiologic significance of the sigma receptors in the various lymphoid and endocrine tissues remains to be determined, but their presence in such high densities suggests that endogenous sigma ligands may play an important role in regulating and integrating endocrine and immune responses. Thus, in addition to their central actions, sigma agonists including PCP and SKF 10,047 may exert their immunosuppressive and endocrine effects directly through actions in the pituitary and target organs. On the other hand, the effects of selective PCP agonists on endocrine function appear to be mediated primarily through actions in brain. Furthermore, the immunologic and endocrine effects of neuroleptics such as haloperidol, which have previously been attributed primarily to actions at D2 dopamine receptors, may also be mediated via sigma receptors in brain, lymphoid, and endocrine organs. Finally, the immune and neuroendocrine systems may represent useful "windows to the brain" to assess the role of sigma and PCP receptors in the CNS. PMID- 8232519 TI - The international conference on PHC. PMID- 8232520 TI - [Organizational climate]. PMID- 8232521 TI - Chronic pain and the pain clinic. PMID- 8232522 TI - Contraceptives for teenagers--an ethical issue. PMID- 8232523 TI - Should AIDS be notifiable? PMID- 8232524 TI - Caring for the cancer patient. PMID- 8232525 TI - The future of psychiatric nursing. PMID- 8232526 TI - Words, idle words. PMID- 8232527 TI - Critics condemn NIH women's study. PMID- 8232528 TI - Protein prenylation. Mad bet for Rab. PMID- 8232530 TI - Structural Biology. PMID- 8232529 TI - Diabetes. Spotlight on a neuronal enzyme. PMID- 8232531 TI - Viral burden in AIDS. PMID- 8232532 TI - Viral burden in AIDS. PMID- 8232533 TI - Skin cancer and UV radiation. PMID- 8232534 TI - NIH awards gene centre grants. PMID- 8232535 TI - The DNA replication fork can pass RNA polymerase without displacing the nascent transcript. AB - Replication proteins encoded by bacteriophage T4 generate DNA replication forks that can pass a molecule of Escherichia coli RNA polymerase moving in the same direction as the fork in vitro. The RNA polymerase ternary transcription complex remains bound to the DNA and retains a transcription bubble after the fork passes. The by-passed ternary complex can resume faithful RNA synthesis, suggesting that the multisubunit RNA polymerase of E. coli has evolved to retain its transcript after DNA replication, allowing partially completed transcripts to be elongated into full-length RNA molecules. PMID- 8232536 TI - Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation. AB - The use of fluorescence microscopy for investigating the three-dimensional structure of cells and tissue is of growing importance in cell biology, biophysics and biomedicine. Three-dimensional data are obtained by recording a series of images of the specimen as it is stepped through the focal plane of the microscope. Whether by direct imaging or by confocal scanning, diffraction effects and noise generally limit axial resolution to about 0.5 microns. Here we describe a fluorescence microscope in which axial resolution is increased to better than 0.05 microns by using the principle of standing-wave excitation of fluorescence. Standing waves formed by interference in laser illumination create an excitation field with closely spaced nodes and antinodes, allowing optical sectioning of the specimen at very high resolution. We use this technique to obtain images of actin fibres and filaments in fixed cells, actin single filaments in vitro and myosin II in a living cell. PMID- 8232537 TI - MRC to limit patents on cDNA sequences. PMID- 8232538 TI - On the molecular origin of photoreceptor noise. AB - Retinal photoreceptors are noisy. They generate discrete electrical events in the dark indistinguishable from those evoked by light and thereby limit visual sensitivity at low levels of illumination. The random spontaneous events are strongly temperature-dependent and have been attributed to thermal isomerizations of the vitamin A chromophore of rhodopsin, the light-sensitive molecule in photoreceptors. But thermal generation of dark events in both vertebrate and invertebrate photoreceptors requires activation energies in the range of 23 to 27 kcal mol-1, which are significantly less than the energy barrier of 45 kcal mol 1, for photoisomerization of the chromophore of native rhodopsin. We propose that photoreceptor noise results from the thermal isomerization of a relatively unstable form of rhodopsin, one in which the Schiff-base linkage between the chromophore and protein is unprotonated. This molecular mechanism is supported by both theoretical calculations of the properties of rhodopsin and experimental measurements of the properties of photoreceptor noise. PMID- 8232539 TI - Immune response to glutamic acid decarboxylase correlates with insulitis in non obese diabetic mice. AB - Knowing the autoantigen target(s) in an organ-specific autoimmune disease is essential to understanding its pathogenesis. Insulin-dependent diabetes mellitus (IDDM) is an autoimmune disease characterized by lymphocytic infiltration of the islets of Langerhans (insulitis) and destruction of insulin-secreting pancreatic beta-cells. Several beta-cell proteins have been identified as autoantigens, but their importance in the diabetogenic process is not known. The non-obese diabetic (NOD) mouse is a murine model for spontaneous IDDM. Here we determine the temporal sequence of T-cell and antibody responses in NOD mice to a panel of five murine beta-cell antigens and find that antibody and T-cell responses specific for the two isoforms of glutamic acid decarboxylase (GAD) are first detected in 4 week-old NOD mice. This GAD-specific reactivity coincides with the earliest detectable response to an islet extract, and with the onset of insulitis. Furthermore, NOD mice receiving intrathymic injections of GAD65 exhibit markedly reduced T-cell proliferative responses to GAD and to the rest of the panel, in addition to remaining free of diabetes. These results indicate that the spontaneous response to beta-cell antigens arises very early in life and that the anti-GAD immune response has a critical role in the disease process during this period. PMID- 8232540 TI - Spanish laboratory backs off bid to cut research staff. PMID- 8232541 TI - The mitotic feedback control gene MAD2 encodes the alpha-subunit of a prenyltransferase. AB - The mad2-1 mutation inactivates the cell-cycle feedback control that prevents budding yeast cells from leaving mitosis until spindle assembly is complete. The gene product of MAD2 shows significant sequence similarity to the alpha-subunit of prenyltransferases. Here we isolate a new temperature-sensitive mad2 mutant, mad2-2ts, and find that Mad2p is required for the membrane association of Ypt1p and Sec4p, two prenylated small GTP-binding proteins involved in protein trafficking. Extracts from mad2-2ts mutant cells fail to geranylgeranylate a number of substrates at the non-permissive temperature. mad2-2ts is synthetically lethal with bet2-1, a mutation in the gene that encodes for the beta-subunit of the Ypt1p and Sec4p geranylgeranyl transferase. Therefore MAD2 and BET2 gene products may physically interact to form a geranylgeranyl transferase complex. In addition, the difference between the phenotypes of mad2-1 and mad2-2ts suggests that MAD2 has distinct roles in protein transport and the mitotic feedback control. PMID- 8232542 TI - Bet2p and Mad2p are components of a prenyltransferase that adds geranylgeranyl onto Ypt1p and Sec4p. AB - Three different prenyltransferases have been identified in yeast and higher cells, the farnesyltransferase and the type I and type II geranylgeranyltransferases (GGTase). The farnesyltransferase and GGTase-I modify peptides in vitro with the CAAX (C, Cys; A, aliphatic residue; X, terminal amino acid) consensus motif. These enzymes are heterodimers that have different beta subunits and a shared alpha-subunit. In yeast, the RAM2 gene encodes this alpha subunit. RAM2 is also homologous to MAD2, a yeast gene whose product has been implicated in the feedback control of mitosis. We have shown that Bet2p is a component of the yeast GGTase-II (refs 6, 12) that geranylgeranylates Ypt1p, a small GTP-binding protein that mediates transport from the endoplasmic reticulum to the Golgi complex. Here we report that Mad2p is a component of this enzyme. Bet2p forms a complex with Mad2p that appears to bind geranylgeranyl pyrophosphate, but not farnesyl pyrophosphate. The efficient transfer of geranylgeranyl onto small GTP-binding proteins requires the presence of an additional activity. PMID- 8232543 TI - Yeast origin recognition complex is involved in DNA replication and transcriptional silencing. AB - The HMR E silencer represses transcription of silent mating-type genes in the budding yeast Saccharomyces cerevisiae and contains three redundant regulatory elements A, E and B (ref. 1). The A element contains the 11 base pair consensus sequence that is essential for the firing of DNA replication origins. A multisubunit protein called the origin recognition complex (ORC) binds specifically to this consensus sequence within yeast origins in vitro and in vivo. We isolated mutants in A element-mediated silencing and report here that one of the genes we identified, RRR1, encodes ORC2, the 72K subunit of ORC. RRR1/ORC2 is an essential gene, but the rrr1-316 allele, which is viable, is defective in the replication of nuclear DNA and the maintenance of the 2-microns episomal DNA. This is, to our knowledge, the first genetic evidence that ORC is involved in DNA replication and silencing. PMID- 8232544 TI - Celltech announces public offering. PMID- 8232545 TI - Genetic testing under scrutiny in US. PMID- 8232546 TI - Slide viewer. PMID- 8232547 TI - Has nature overwhelmed nurture? PMID- 8232548 TI - Plant biology. New light on stem growth. PMID- 8232549 TI - Phototransduction. The invertebrate enigma. PMID- 8232550 TI - Signal transduction. Cytokine connections. PMID- 8232551 TI - X larger than Y. PMID- 8232552 TI - The protein tyrosine kinase JAK1 complements defects in interferon-alpha/beta and -gamma signal transduction. AB - We have produced a cell line which lacks the protein tyrosine kinase JAK1 and is completely defective in interferon response. Complementation of this mutant with JAK1 restored the response, establishing the requirement for JAK1 in both the interferon-alpha/beta and -gamma signal transduction pathways. The reciprocal interdependence between JAK1 and Tyk2 activities in the interferon-alpha pathway, and between JAK1 and JAK2 in the interferon-gamma pathway, may reflect a requirement for these kinases in the correct assembly of interferon receptor complexes. PMID- 8232553 TI - Episodic multiregional cortical coherence at multiple frequencies during visual task performance. AB - The way in which the brain integrates fragmentary neural events at multiple locations to produce unified perceptual experience and behaviour is called the binding problem. Binding has been proposed to involve correlated activity at different cortical sites during perceptuomotor behaviour, particularly by synchronization of narrow-band oscillations in the gamma-frequency range (30-80 Hz). In the rabbit olfactory system, inhalation induces increased gamma correlation between sites in olfactory bulb and cortex. In the cat visual system, coherent visual stimuli increase gamma-correlation between sites in both the same and different visual cortical areas. In monkeys, some groups have found that gamma-oscillations transiently synchronize within striate cortex, superior temporal sulcus and somatosensorimotor cortex. Others have reported that visual stimuli produce increased broad-band power, but not gamma-oscillations, in several visual cortical areas. But the absence of narrow-band oscillations in itself does not disprove interregional synchronization, which may be a broad-band phenomenon. We now describe episodes of increased broad-band coherence among local field potentials from sensory, motor and higher-order cortical sites of macaque monkeys performing a visual discrimination task. Widely distributed sites become coherent without involving other intervening sites. Spatially selective multiregional cortical binding, in the form of broad-band synchronization, may thus play a role in primate perceptuomotor behaviour. PMID- 8232554 TI - Molecular determinants of Ca2+ selectivity and ion permeation in L-type Ca2+ channels. AB - Voltage-gated Ca2+ channels link changes in membrane potential to the delivery of Ca2+, a key second messenger for many cellular responses. Ca2+ channels show selectivity for Ca2+ over more plentiful ions such as Na+ or K+ by virtue of their high-affinity binding of Ca2+ within the pore. It has been suggested that this binding involves four conserved glutamate residues in equivalent positions in the putative pore-lining regions of repeats I-IV in the Ca2+ channel a1 subunit. We have carried out a systematic series of single amino-acid substitutions in each of these positions and find that all four glutamates participate in high-affinity binding of Ca2+ or Cd2+. Each glutamate carboxylate makes a distinct contribution to ion binding, with the carboxylate in repeat III having the strongest effect. Some single glutamate-to-lysine mutations completely abolish micromolar Ca2+ block, indicating that the pore does not possess any high affinity binding site that acts independently of the four glutamate residues. The prevailing model of Ca2+ permeation must thus be modified to allow binding of two Ca2+ ions in close proximity, within the sphere of influence of the four glutamates. The functional inequality of the glutamates may be advantageous in allowing simultaneous interactions with multiple Ca2+ ions moving single-file within the pore. Competition among Ca2+ ions for individual glutamates, together with repulsive ion-ion electrostatic interaction, may help achieve rapid flux rates through the channel. PMID- 8232555 TI - HY4 gene of A. thaliana encodes a protein with characteristics of a blue-light photoreceptor. AB - Specific responses to blue light are found throughout the biological kingdom. These responses--which in higher plants include phototropism, inhibition of hypocotyl elongation, and stomatal opening--are in many cases thought to be mediated by flavin-type photoreceptors. But no such blue-light photoreceptor has yet been identified or isolated, although blue-light responses in plants were reported by Darwin over a century ago, long before the discovery of the now relatively well characterized red/far-red light photoreceptor, phytochrome. Here we describe the isolation of a gene corresponding to the HY4 locus of Arabidopsis thaliana. The hy4 mutant is one of several mutants that are selectively insensitive to blue light during the blue-light-dependent inhibition of hypocotyl elongation response, which suggests that they lack an essential component of the cryptochrome-associated light-sensing pathway. The HY4 gene, isolated by gene tagging, was shown to encode a protein with significant homology to microbial DNA photolyases. As photolyases are a rare class of flavoprotein that catalyse blue light-dependent reactions, the protein encoded by HY4 has a structure consistent with that of a flavin-type blue-light photoreceptor. PMID- 8232556 TI - BRG1 contains a conserved domain of the SWI2/SNF2 family necessary for normal mitotic growth and transcription. AB - Sequence-specific DNA binding activators of gene transcription may be assisted by SWI2 (SNF2), which contains a DNA-dependent ATPase domain. We have isolated a human complementary DNA encoding a 205K nuclear protein, BRG1, that contains extensive homology to SWI2 and Drosophila brahma. We report here that a SWI2/BRG1 chimera with the DNA-dependent ATPase domain replaced by corresponding human sequence restored normal mitotic growth and capacity for transcriptional activation to swi2- yeast cells. Point mutation of the conserved ATP binding site lysine abolished this complementation. This mutation in SWI2 exerted a dominant negative effect on transcription in yeast. A lysine to arginine substitution at the corresponding residue of BRG1 also generated a transcriptional dominant negative in human cells. BRG1 is exclusively nuclear and present in a high M(r) complex of about 2 x 10(6). These results show that the SWI2 family DNA-dependent ATPase domain has functional conservation between yeast and humans and suggest that a SWI/SNF protein complex is required for the activation of selective mammalian genes. PMID- 8232557 TI - Transposition of group II intron aI1 in yeast and invasion of mitochondrial genes at new locations. AB - Intron mobility at the RNA level by splicing reversal at allelic (homing) and non allelic locations (transposition) has been reported in vitro. In the living cell, however, only intron homing by unidirectional gene conversion has been described. Supposing that intron insertions at non-allelic sites might occur in vivo, we speculated that group II splice-site-associated macro-deletions in fungal mitochondrial DNA might result from group II intron transposition to new locations followed by recombination. We used polymerase chain reaction techniques to detect this critical, infrequent intermediate in mtDNA populations. Here we report on group II intron aI1 transposition to non-allelic, splicing-compatible locations within the cox1 gene of yeast mtDNA. The identified integration sites are preceded by motifs similar to the upstream exon A1. Sequences flanking intron aI1 are not co-converted to the insertion sites and cis- and trans-acting mutations within aI1 reduce intron mobility below detection levels. These findings suggest the involvement of an RNA intermediate in group II intron transposition. PMID- 8232558 TI - Transposition of a group II intron. AB - Among mobile genetic elements, self-splicing introns are of particular interest. They belong to either group I or group II depending on their three-dimensional structure. Homing, the systematic intron invasion of an intronless gene when it encounters its homologous intron-bearing allele, is the only means for intron mobility so far demonstrated. It depends on the activity of the intron-encoded protein and is very specific for the acceptor site. Intron transposition, the transfer of an intron to a novel site, predicted on the basis of phylogenetic studies and in vitro reverse-splicing experiments, has been proposed to be responsible for evolutionary intron spreading. Here we present results from polymerase chain reaction experiments consistent with transposition of a group II intron. This event is proposed to account for the site-specific deletion in the mitochondrial chromosome of the fungus Podospora anserina that is associated with the premature death syndrome and might also be involved in the senescence process affecting this species. PMID- 8232559 TI - Tandem binding in crystals of a trp repressor/operator half-site complex. AB - The crystal structure of trp repressor tandemly bound in a 2:1 complex to a 16 base-pair palindromic DNA containing a central trp operator half-site has been determined and refined to 2.4 A resolution. Despite dramatically different DNA sequence contexts and crystallization conditions, the protein/DNA interface is essentially identical to that seen in the original trp repressor/operator complex structure. Water-mediated sequence recognition by trp repressor is likely to be related to the unusual end-on approach of the recognition helix (E), which allows sharing of the major groove by tandem dimers. The tandem complex model accounts for the mutational sensitivity of all trp operator base pairs. The structure also provides the first detailed view of the tandem interaction, revealing a key role for the amino-terminal arms. PMID- 8232561 TI - Europe's HIV-contaminated blood. PMID- 8232560 TI - New ideas for personal library maintenance software. PMID- 8232562 TI - An end to fraud? PMID- 8232563 TI - Concern grows over impact of German blood scandal. PMID- 8232564 TI - Creutzfeldt-Jakob verdict may prompt new claims. PMID- 8232565 TI - Paracelsus. PMID- 8232566 TI - Popovic wins appeal against ORI charges. PMID- 8232567 TI - Varmus deft with senate on NIH strategy. PMID- 8232568 TI - French keeps up claims to AIDS patent. PMID- 8232569 TI - Misconduct charges against Gallo withdrawn after Popovic decision. PMID- 8232570 TI - Milk hormone faces new hurdles on way to market. PMID- 8232571 TI - Protest group turns spotlight on university researcher. PMID- 8232572 TI - UK rules out new gene therapy laws. PMID- 8232573 TI - UNESCO and copyright. PMID- 8232574 TI - Health research feels the chill. AB - Reforms in the US system for financing health care have grave implications for academic medicine and for basic scientific research. PMID- 8232575 TI - Genetics. Small and perfectly formed. PMID- 8232576 TI - Why we still love Lucy. PMID- 8232577 TI - Graphology. The writing is on the wall. PMID- 8232578 TI - Cell cycle. Sharper than a needle. PMID- 8232579 TI - Lifespan and testosterone. PMID- 8232580 TI - Muzzle length and heat loss. PMID- 8232581 TI - Viral extinctions in deep-sea species. PMID- 8232582 TI - Punctuated equilibrium comes of age. AB - The intense controversies that surrounded the youth of punctuated equilibrium have helped it mature to a useful extension of evolutionary theory. As a complement to phyletic gradualism, its most important implications remain the recognition of stasis as a meaningful and predominant pattern within the history of species, and in the recasting of macroevolution as the differential success of certain species (and their descendants) within clades. PMID- 8232583 TI - Intrinsic density-dependent regulation of vole populations. AB - Considerable controversy exists over the role of density-dependent processes in controlling animal population size. In populations that fluctuate cyclically or erratically, for example many voles and insects, theory predicts that either density-dependence is weak, or that density-dependent responses lag behind density. One key mechanism for lagged density-dependence is a delay in regeneration of food resources following heavy exploitation. Here we show that meadow vole (Microtus pennsylvanicus) populations respond immediately to high density by reducing breeding effort and hence population growth, disproving the hypothesis that density-dependence is weak. In addition, vole populations do not show a delay in growth following marked reduction in plant biomass (their source of food and cover). We conclude that intrinsic density-dependence processes tend to stabilize vole populations, and that cyclic dynamics are not caused by lagged effects of resource exploitation. PMID- 8232584 TI - New discoveries of Australopithecus at Maka in Ethiopia. AB - The taxonomy of Australopithecus afarensis, the oldest known hominid species, has been a matter of debate since its description in 1978 (ref. 1). Some authorities regard all specimens assigned to A. afarensis as belonging to a single taxon whereas others regard the Tanzanian and Ethiopian specimens as each representing a different species. Further controversy surrounds the issues of sexual dimorphism and locomotion among these hominids. Resolution of these problems would shed light on hominid phylogeny in general and on the ancestry of later Australopithecus and Homo. Fossils discovered in the Afar of Ethiopia in 1990 constitute the first major addition to the 3-4 million year (Myr) hominid record since the 1970s. We report here the discovery of new fossils from Maka, dated to 3.4 Myr ago, which provide powerful support for the interpretation of A. afarensis as a single, ecologically diverse, sexually dimorphic, bipedal Pliocene primate species whose known range encompassed Ethiopia and Tanzania. PMID- 8232585 TI - Characterization of the pufferfish (Fugu) genome as a compact model vertebrate genome. AB - Cloning and sequencing techniques now allow us to characterize genes directly instead of having to deduce their properties from their effects. This new genetics reaches its apotheosis in the plan to obtain the complete DNA sequence of the human genome, but this is far beyond the capacity of present sequencing methods. Small 'model' genomes, 'such as those of Escherichia coli (4.7 megabases (Mb) and yeast (14 Mb), or even those of Caenorhabditis elegans (100 Mb) and Drosophila (165 Mb), are better scaled to existing technology. The yeast genome will contain genes with functions common to all eukaryotic cells, and those of simple multicellular organisms may throw light on the genetic specification of more complex functions. However, vertebrates differ in their morphology and development, so the ideal model would be a vertebrate genome of minimum size and complexity but with maximum homology to the human genome. Here we report the characterization of the small genome (400 Mb) of the tetraodontoid fish, Fugu rubripes. A random sequencing approach supported by gene probing shows that the haploid genome contains 400 Mb of DNA, of which more that 90% is unique. This genome is 7.5 times smaller than the human genome and because it has a similar gene repertoire it is the best model genome for the discovery of human genes. PMID- 8232586 TI - Novel heterotrimeric kinesin-related protein purified from sea urchin eggs. AB - Kinesin heavy chain and kinesin-related polypeptides (KRPs) comprise a family of motor proteins with diverse intracellular transport functions. Using pan-kinesin peptide antibodies that react with these proteins, we have previously purified from sea urchin eggs a trimeric microtubule-binding and bundling protein, KRP (85/95) (ref. 8) comprising subunits of M(r) 115,000 (115K), 95K and 85K. We report here that kinesin-related genes encode the 85K and 95K subunits, and that the protein can be immunoprecipitated from cytosol as a trimeric complex using an 85K monoclonal antibody. We also find that purified KRP(85/95) directs movements towards the 'plus' ends of microtubules. To our knowledge, this protein is the first kinesin-related motor to be purified from its natural host cell in a native multimeric state. PMID- 8232587 TI - Calmodulin-dependent protein kinase II mediates inactivation of MPF and CSF upon fertilization of Xenopus eggs. AB - In vertebrates, unfertilized eggs are arrested at second meiotic metaphase by a cytostatic factor (CSF), an essential component of which is the product of the c mos proto-oncogene. CSF prevents ubiquitin-dependent degradation of mitotic cyclins and thus inactivation or the M phase-promoting factor (MPF). Fertilization or parthenogenetic activation triggers a transient increase in the cytoplasmic free Ca2+ (reviewed in refs 5 and 6), inactivates both CSF and MPF, and releases eggs from meiotic metaphase arrest. A calmodulin-dependent process is required for cyclin degradation to occur in cell-free extracts prepared from metaphase II-arrested eggs (CSF extracts) when the free Ca2+ concentration is transiently raised in the physiological micromolar range. Here we show that when a constitutively active mutant of calmodulin-dependent protein kinase II (CaM KII) is added to a CSF extract, cyclin degradation and Cdc2 kinase inactivation occur even in the absence of Ca2+, and the extract loses its ability to cause metaphase arrest when transferred into embryos. Furthermore, specific inhibitors of CaM KII prevent cyclin degradation after calcium addition. Finally, the direct microinjection of constitutively active CaM KII into unfertilized eggs inactivates Cdc2 kinase and CSF, even in the absence of a Ca2+ transient. The target for Ca(2+)-calmodulin is thus CaM KII. PMID- 8232588 TI - Bcl-2 associates with the ras-related protein R-ras p23. AB - Apoptosis is an important but poorly understood mechanism of cell regulation. Growth factor deprivation can trigger apoptosis in a variety of cells, suggesting the existence of a signal transduction pathway responding to external signals and leading to apoptosis. Overexpression of the proto-oncogene bcl-2 can override these signals and block apoptosis, indicating that the bcl-2 protein (Bcl-2) is an important component of the apoptotic response. The identification of Bcl-2 binding proteins might help explain how Bcl-2 acts to regulate apoptosis. Here we use the yeast two-hybrid system to show that the human ras-related protein R-ras p23 (ref 16-18) binds to Bcl-2. This association is also detected in immunoprecipitates from human cell extracts. The association requires full-length Bcl-2 but the C-terminal 60 amino acids of R-ras p23 are sufficient for the interaction. These results provide evidence of a putative component of a signal transduction pathway involved in the regulation of apoptosis. PMID- 8232589 TI - Cloning and expression of cytochrome P450 genes controlling flower colour. AB - Blue and violet flowers generally contain derivatives of delphinidin; red and pink flowers generally contain derivatives of cyanidin or pelargonidin. Differences in hydroxylation patterns of these three major classes of anthocyanidins are controlled by the cytochrome P450 enzymes flavonoid 3' hydroxylase and flavonoid 3',5'-hydroxylase. Here we report on the isolation of complementary DNA clones of two different flavonoid 3',5'-hydroxylase genes that are expressed in petunia flowers. Restriction-fragment length polymorphism mapping and complementation of mutant petunia lines showed that the flavonoid 3',5'-hydroxylase genes correspond to the genetic loci Hf1 and Hf2. PMID- 8232590 TI - Nonlinear amplification by calcium-dependent chloride channels in olfactory receptor cells. AB - The sense of smell is highly evolved in mammals, allowing discrimination between a vast number of odorants, with detection thresholds as low as 10(-17) M (ref. 1). Although several features of mammalian olfactory transduction have been revealed by biochemical and molecular biological studies, the odorant-induced membrane current has remained elusive. In amphibians this current is mediated by cyclic-nucleotide-gated channels, which depolarize the cell by Na+ and Ca+ influx and consequent Cl- efflux through Ca(2+)-dependent Cl- channels. The Cl- current may be absent in mammals, however, because its proposed role is linked to the aquatic habitat of amphibians. Here we show that the transduction current in rat olfactory receptor cells is initiated by cyclic-nucleotide-gated channels. The Cl current is also present and endows the transduction current with a steep sigmoidal dependence on cyclic AMP concentration in both rat and in an amphibian, indicating a new function for the Cl- channel: nonlinear amplification of the transduction signal, whereby suprathreshold responses are boosted relative to basal transduction noise. PMID- 8232591 TI - Electrically induced release of endogenous noradrenaline and dopamine from brain slices: pseudo-one-pulse stimulation utilized to study presynaptic autoinhibition. AB - Slices of rat hypothalamus (noradrenaline experiments) or rabbit caudate nucleus (dopamine experiments) were prepared, superfused, and field-stimulated using series of monophasic rectangular pulses. Noradrenaline, dopamine and the main dopamine metabolite, dihydroxyphenylacetic acetic acid (DOPAC), were determined using HPLC with electrochemical detection. Electrical stimulation was performed using the following protocols: 1) 4 pulses delivered at 100 Hz; this type of stimulation is referred to as pseudo-one-pulse stimulation (POP); its short duration of only 32 ms does not allow the development of autoinhibition; 2) 2 bursts of 4 pulses at 100 Hz, delivered 1 s apart (2-POP-stimulation); 3) 8 pulses at 1 Hz (dopamine experiments only); 4) 36 pulses at 3 Hz. Noradrenaline experiments. The alpha 2-adrenoceptor antagonist yohimbine (1 mumol/l) did not enhance noradrenaline overflow following POP stimulation, but enhanced the overflow following 2-POP-stimulation by about 50% and that following 36-pulse stimulation by almost 100%. Dopamine experiments. The D2-dopamine receptor antagonist sulpiride (3 mumol/l) facilitated the overflow of dopamine elicited with 2-POP-stimulation (66%), 8 pulses/1 Hz (92%), and 36 pulses/3 Hz (140%). It did not significantly facilitate the overflow of dopamine following POP stimulation (19%). The overflow of DOPAC was not, or only slightly, increased by electrical stimulation, and its spontaneous outflow was more than three times higher than that of dopamine. Furthermore, the electrically induced overflow of dopamine did not exceed the outflow of DOPAC at any of the stimulation conditions employed. The results of the present study bear out important claims of the autoreceptor theory and confirm the data obtained in previous experiments using labelled transmitters. PMID- 8232592 TI - Prejunctional inhibition of sympathetically evoked pupillary dilation in cats by activation of histamine H3 receptors. AB - Frequency-dependent pupillary dilations were evoked by electrical stimulation of the pre- or post-ganglionic cervical sympathetic nerve (sympatho-excitation) or the hypothalamus (parasympatho-inhibition) in sympathectomized anesthetized cats. Systemic administration of the selective histamine H3 receptor agonist (R)-alpha methylhistamine (R alpha MeHA) produced a dose-dependent depression of mydriasis due to direct neural sympathetic activation but had no effect on responses elicited by parasympathetic withdrawal. The histamine H2 receptor agonist, dimaprit, was inactive. R alpha MeHA was much more effective in depressing sympathetic responses obtained at lower frequencies when compared to higher frequencies of stimulation. Responses evoked both pre- and postganglionically were inhibited by R alpha MeHA. This peripheral sympatho-inhibitory action of R alpha MeHA was antagonized by the histamine H3 receptor blocker thioperamide but not by intravenous pretreatment with the histamine H1 receptor antagonist chlorpheniramine. Histamine H2 receptor blockers cimetidine and ranitidine were also without effect. R alpha MeHA did not depress pupillary responses elicited by i.v. (-)-adrenaline. The results demonstrate that histamine H3 receptors modulate sympathetic activation of the iris at a site proximal to the iris dilator muscle. The predominant mechanism of action appears to the prejunctional inhibition of noradrenaline release from postganglionic sympathetic nerve endings. However, a concomitant ganglionic inhibitory action cannot be excluded. PMID- 8232593 TI - Involvement of both GABAA and GABAB receptors in tonic inhibitory control of blood pressure at the rostral ventrolateral medulla of the rat. AB - The rostral ventrolateral medulla (RVLM) contains vasopressor neurons which increase vasomotor tone. Endogenous GABA is suggested to be involved in mediation of the tonic inhibition of vasopressor neurons in the RVLM. To obtain more precise information about GABAergic mechanisms in the RVLM, we microinjected GABA agonists and antagonists unilaterally into the RVLM and examined their effects on blood pressure and heart rate. In addition, involvement of the other inhibitory amino acids glycine, beta-alanine and taurine in blood pressure regulation in the rat RVLM was also investigated. Male Wistar rats were anesthetized with urethane, paralyzed and artificially ventilated. The GABAA agonist muscimol (3-30 pmol) and the GABAB agonist baclofen (10-100 pmol) microinjected into the RVLM produced a decrease in blood pressure. The GABAA antagonist bicuculline (300 pmol) abolished the depressor response to muscimol (10 pmol) but not to baclofen (30 pmol) whereas the GABAB antagonist 2-hydroxysaclofen (1 nmol) abolished the depressor response to baclofen (30 pmol) but not to muscimol (10 pmol). Either bicuculline or 2-hydroxysaclofen alone produced a pressor response. Both antagonists inhibited depressor responses to nipecotic acid (7.7 nmol) and GABA (0.3 nmol). Glycine (0.13-4.0 nmol), beta-alanine (0.11-3.4 nmol) and taurine (0.08-2.4 nmol) microinjected into the RVLM also produced decreases in blood pressure. The glycine antagonist strychnine (0.58 nmol) abolished the depressor response to glycine, beta-alanine and taurine but not to GABA. The taurine antagonist 6 aminomethyl-3-methyl-4H-1,2,4-benzothiadiazine-1,1-dioxide) (1.3 nmol) inhibited the depressor response to beta-alanine and taurine but not to glycine and GABA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232594 TI - Possible explanations for the antagonism by nicotine against reserpine-induced depletion of monoamines in mouse brain. AB - The inhibitory effect of nicotine pretreatment on reserpine-induced depletion of monoamines in mouse brain was investigated. The depletion of brain monoamines by 24 h after intraperitoneal injection of reserpine (2 mg/kg) was dose-dependently inhibited by nicotine (0.3-10 mg/kg, s.c.) pretreatment 20 min before reserpine injection. This effect of nicotine was more marked on dopamine depletion than on noradrenaline or 5-hydroxytryptamine depletion. The nicotine pretreatment also inhibited the reserpine-induced hypothermia and decrease in the locomotor activity. When reserpine (2 mg/kg) was injected intraperitoneally, the inhibitory effect of nicotine (3 mg/kg, s.c.) on the reserpine-induced depletion of brain monoamines and heart noradrenaline was not antagonized by hexamethonium (8 mg/kg, s.c.) but rather potentiated by mecamylamine (2 mg/kg, s.c.). However, when reserpine (0.5 mg/kg) was injected intravenously, pretreatment with nicotine (3 mg/kg, s.c.) inhibited the reserpine-induced dopamine depletion only, and this effect of nicotine was completely blocked by mecamylamine but not by hexamethonium. These results suggest that inhibitory effect of nicotine on the intraperitoneal reserpine-induced depletion of brain monoamines is due to an inhibition of absorption of reserpine, and that central nicotinic action is also involved in the antagonism by nicotine of reserpine-induced dopamine depletion. PMID- 8232595 TI - Use-dependent effects of acute and chronic treatment with imipramine and buspirone on excitatory synaptic transmission in the rat hippocampus in vivo. AB - The effects of acute and long-term treatment with imipramine and buspirone on the responses of rat hippocampal neurones to low and high frequency electrical stimulation were compared. Whereas acute treatment with imipramine (10 mg/kg, i.p.) had no effect on synaptic responses to low frequency stimulation, chronic treatment for 14 days significantly reduced the amplitude of the field excitatory postsynaptic potential. Both acute and chronic imipramine treatment markedly reduced the amplitude of the nerve volley and excitatory postsynaptic potential evoked at high frequency stimulation rates in a use-dependent manner. Buspirone (0.5-3 mg/kg, i.p.) produced a significant reduction of the excitatory postsynaptic potential at high frequencies. This was enhanced after repeated administration of a dose of 0.5 mg/kg for 14 days. We previously reported a similar effect of buspirone at low frequency stimulation. Both compounds therefore share the ability to exert strong depressant effects on transmission in the hippocampus especially after chronic treatment. PMID- 8232596 TI - Pharmacological characterisation of neurokinin receptors mediating anion secretion in rat descending colon mucosa. AB - Substance P (SP), neurokinin A (NKA), neurokinin B (NKB), [Sar9, Met (O2)11]-SP (SMSP), senktide, [beta Ala8]-NKA(4-10) and neuropeptide gamma (NP gamma) all stimulate secretory responses in rat descending colon mucosa under voltage clamp conditions. Secretory responses (measured as short circuit current under voltage clamp conditions) were transient and those evoked by SP, SMSP, NKA and senktide were significantly reduced by pretreating tissues with the chloride channel blocker, diphenylamine carboxylate (DPC). Concentration-response curves showed varying degrees of sensitivity to tetrodotoxin (TTX). Senktide-induced secretion was virtually abolished by TTX, while NP gamma and [beta Ala8]-NKA(4-10) were not significantly altered. Rightward shifts of concentration-response curves were observed for SMSP, NKA and SP in TTX treated preparations compared with controls. NKA response curves in the presence of TTX were further inhibited by MEN10,207 and CP-96,345. GR71251, GR82334 and CP-96,345 all inhibited SMSP secretory responses with pA2 values of 5.8, 6.5 and 6.9 respectively. In conclusion three types of neurokinin receptor exist in preparations of rat colon mucosa and their relative location within neuronal and epithelial surfaces are discussed. PMID- 8232597 TI - Effect of LP-805, a releaser of endothelium-derived nitric oxide, on systemic vasodilatation in vivo. AB - We have investigated relations between hypotensive responses to LP-805, a newly synthesized vasodilator, and the production of nitric oxide (NO), in anesthetized rats. LP-805 (0.1-0.5 mg/kg, i.v.) or acetylcholine (ACh) (0.3-3.0 micrograms/kg, i.v.) caused a dose-dependent transient decrease in diastolic blood pressure. The decrease induced by 0.3 mg/kg LP-805 (i.v.) was partially inhibited by pretreatment with NG-nitro-L-arginine (L-NNA), a specific inhibitor of endothelial NO synthase, but the responses to lower or higher doses of LP-805 (0.1 or 0.5 mg/kg, i.v.) were not affected. The dose-dependent decrease in diastolic blood pressure, caused by LP-805, was not affected by pretreatment with L- or D-arginine. The dose-dependent decrease in diastolic blood pressure caused by ACh was not affected by pretreatment with L-NNA or with L- or D-arginine. The hypotensive response to 20-min infusions of LP-805 (100 micrograms/kg per min) was significantly inhibited by pretreatment with L-NNA (10 mg/kg, i.v.). The half recovery times (T 1/2) of LP-805 or ACh-induced depressor responses were shortened by pretreatment with L-NNA. They were prolonged by L-arginine, but not by D-arginine. This shortening, by L-NNA, of the half-recovery time after LP-805 or ACh was reversed by L-arginine, but not by D-arginine. The T 1/2 of the LP-805 induced hypotensive response was not affected by pretreatment with indomethacin (1 mg/kg, i.v.). In the presence of L-NNA (10 mg/kg, i.v.), the T 1/2 of the LP 805-induced hypotensive response was not affected by pretreatment with indomethacin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232598 TI - Virus-induced airway inflammation and hyperresponsiveness in the guinea-pig is inhibited by levodropropizine. AB - Intratracheal Parainfluenza type 3 (PI-3) virus inoculation of guinea pigs leads to a non-specific airway hyperresponsiveness in vivo and in vitro which coincides with a significant increase in the number of inflammatory cells in the broncho alveolar lavage fluid (90% increase, 4 days after inoculation). The activity of the bronchoalveolar cells, as measured by the chemiluminescence production of infected animals is significantly diminished (34.2%, 4 days after inoculation) after renewed stimulation with PI-3 virus in vitro as compared to the chemiluminescence production by bronchoalveolar cells obtained from control guinea pigs. Pretreatment of the guinea-pigs with the antitussive agent levodropropizine, administered intra-peritoneally twice a day for five successive days at a dose of 10 mg/kg, prevents the virus-induced airway hyperresponsiveness in vivo and in vitro, and inhibits the influx of broncho-alveolar cells. Levodropropizine at a dose of 1 mg/kg did not modulate these responses. Further, the decrease in chemiluminescence production of broncho-alveolar cells obtained from virus-infected animals after PI-3 virus stimulation in vitro was inhibited by levodropropizine (10 mg/kg). These data demonstrate the ability of levodropropizine to counteract the hyperresponsiveness phenomenon and the associated inflammatory event induced by PI-3 virus, an effect which may be due to its capacity to act on the peptidergic system or may be due to the anti allergic/bronchoconstrictor property of this compound. PMID- 8232599 TI - The antagonist actions of WAY-100135 and its enantiomers on 5-HT1A receptor mediated hyperpolarization of the rat isolated superior cervical ganglion. AB - A grease-gap extracellular recording technique was used to detect 5-HT1A receptor mediated hyperpolarizing responses to 5-hydroxytryptamine (5-HT) in the rat isolated superior cervical ganglion. In the presence of the novel 5-HT1A receptor antagonist, WAY-100135 [N-tert-butyl-3-(4-(2-methoxyphenyl)piperazin-1-yl)-2 phenylpropan amide], the responses to 5-HT were antagonised in a competitive manner with a pA2 value of 7.2 (6.9-8.5) and Schild plot slope of 1.0 (0.4-1.6), n = 20. The antagonist activity was greater in the (+) than the (-)enantiomer of WAY-100135. The pA2 value of the (+)enantiomer was 7.5 (7.2-8.0), Schild plot slope 1.2 (0.8-1.6), n = 17. In contrast the (-)enantiomer had weak antagonist activity (pA2 6.3 +/- 0.25, n = 3). No agonist activity of WAY-100135 or its enantiomers were observed in this study. PMID- 8232600 TI - Presynaptic muscarinic receptors and the release of acetylcholine from cerebrocortical prisms: roles of Ca2+ and K+ concentrations. AB - The mechanism by which presynaptic muscarinic autoreceptors inhibit the release of acetylcholine (ACh) from cerebrocortical cholinergic fibres has not been clarified. To test the view that muscarinic autoreceptors act by decreasing Ca2+ influx, we performed experiments in which rat cerebrocortical prisms were preloaded with (14C)choline, washed, depolarized with 14-65 mM K+ in the absence of Ca2+ and then exposed (still under depolarization) to various concentrations of Ca2+ to evoke the release of (14C)ACh. The muscarinic agonist, oxotremorine, used at a 100 microM concentration, inhibited the release of (14C)ACh by 59-86% in experiments with 14 and 26.5 mM K+ but had no significant effect at 65.5 mM K+. No systematic changes in the inhibitory effects of oxotremorine could be found at any of the K+ concentrations used when the concentration of Ca2+ was varied in the range of 0.25-4.0 mM. At 2 mM Ca2+ and K+ concentrations above 14 mM, the inhibitory effect of oxotremorine was inversely related to the concentration of K+. The inhibitory effect of oxotremorine on (14C)ACh release was not blocked by 100 microM 4-amino-pyridine. The fact that the inhibitory effect of oxotremorine could not be overcome by an increase in the concentration of Ca2+ suggests that, under the conditions used, a restriction of the influx of Ca2+ did not play a major role in the muscarinic inhibition of ACh release; rather, oxotremorine appeared to act by decreasing membrane depolarization.2+ of the Ca(2+)-voltage hypothesis of neurotransmitter release, supposing PMID- 8232601 TI - No evidence for presynaptic opioid receptors on cholinergic, but presence of kappa-receptors on dopaminergic neurons in the rabbit caudate nucleus: involvement of endogenous opioids. AB - The effects of various opioid receptor agonists and antagonists were studied in rabbit caudate nucleus slices preincubated with either [3H]dopamine or [3H]choline, superfused with medium (containing in most experiments the D2 receptor antagonist domperidone) and subjected to electrical field stimulation. The stimulation-evoked [3H]overflow from slices prelabeled with [3H]dopamine (evoked [3H]dopamine release) was significantly reduced by preferential kappa opioid receptor agonists, like U-50,488 H, but not by mu- or delta-opioid receptor selective drugs. Opioid receptor antagonists shifted the concentration/response curve of U-50,488 H to the right (apparent pA2-value of the kappa-selective antagonist nor-binaltorphimine: 10.1) and enhanced the evoked dopamine release in the presence of a mixture of peptidase inhibitors. On the other hand, the [3H]overflow from rabbit caudate nucleus slices prelabeled with [3H]choline (evoked acetylcholine release) remained almost unaffected by any opioid receptor agonist, as long as the presynaptic D2 heteroreceptor was blocked with domperidone: in the absence of domperidone, U-50,488 H exhibited facilitatory effects. For comparison, the effects of the preferential delta opioid receptor agonist DPDPE was also studied in slices of the rat striatum, where it clearly inhibited the evoked acetylcholine release. From our data we conclude that in the rabbit caudate nucleus the evoked dopamine release is inhibited by both exogenous and endogenous opioids via presynaptic kappa-opioid receptors, whereas the evoked release of acetylcholine is not, or only indirectly (via released dopamine) affected by opioids. PMID- 8232602 TI - Inhibition of uptake 1 by (+)-oxaprotiline reveals a differential central regulation of noradrenaline and adrenaline release. AB - Inhibition of uptake 1 in the central nervous system leads to a decrease of sympathetic outflow to many tissues; central alpha 2-adrenoceptors are involved in this decrease. The aim of the present study was to compare the effects of the selective uptake 1 inhibitor (+)-oxaprotiline on the plasma kinetics of noradrenaline and adrenaline in anaesthetized and in conscious rabbits. [3H]Noradrenaline and [3H]adrenaline were infused i.v. The arterial plasma concentrations of endogenous and radiolabelled noradrenaline and adrenaline were measured, and the clearance from and spillover into the plasma of noradrenaline and adrenaline were calculated. Results obtained in conscious and anaesthetized rabbits were similar. (+)-Oxaprotiline 0.2, 0.6 and 1.8 mg kg-1 i.v. dose dependently reduced the clearance of [3H]noradrenaline from the plasma. The clearance of [3H]adrenaline was reduced less. The spillover of endogenous noradrenaline was decreased by up to 35%. In contrast, the spillover of adrenaline tended to be enhanced. Prazosin 0.1 and 1 mg kg-1 was injected i.v. in a second part of each experiment. It lowered the blood pressure and caused a marked increase in noradrenaline spillover but no increase or even a decrease in adrenaline spillover. The results are compatible with the following hypothesis. The sympathetic outflow from the central nervous system is subject to a twofold alpha-adrenoceptor-mediated modulation: alpha 2-adrenoceptor-mediated inhibition and alpha 1-adrenoceptor-mediated excitation. In the control of the sympathetic outflow to many extra-adrenal tissues, the alpha 2-adrenergic inhibition prevails. Uptake 1 inhibitors depress sympathetic outflow to such tissues by enhancing the alpha 2-adrenergic inhibition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232603 TI - Antagonistic effects of stimulation of the paramedian reticular nucleus in the rat medulla oblongata and of amphetamine on locomotor activity and striatal release of dopamine-like material. AB - The effects of stimulation of the paramedian reticular nucleus (PRN) in the rat medulla oblongata on both amphetamine-induced locomotor activity and striatal release of dopamine-like material were assessed. PRN stimulation (by intra-PRN injection of the excitatory amino acid, kainic acid) decreased vertical motion and total distance travelled, and increased postural freezing, in freely moving rats. On the other hand, a small dose (e.g. 1.25 mg/kg, i.p.) of amphetamine increased locomotor activity (including horizontal motion, vertical motion, total distance travelled and lines crossed counts), increased the number of turnings (both clockwise and anti-clockwise), induced locomotor stereotypy (including both gamma value and number of trip types), and inhibited postural freezing. The changes in activity induced by amphetamine administration were suppressed following PRN stimulation. In vivo voltammetric data revealed that electrical stimulation of the PRN decreased the release of dopamine-like material in the corpus striatum. This effect could be mimicked by intra-PRN injection of kainic acid in anesthetized rats. In contrast, i.p. administration of amphetamine increased the release of dopamine-like material in the corpus striatum. Furthermore, the enhanced release of dopamine-like material induced by amphetamine was attenuated by simultaneous stimulation of the PRN. The results reported here indicate that PRN stimulation decreases the striatal dopamine release and results in attenuation of the amphetamine-induced locomotor activity responses in rats. PMID- 8232604 TI - Effects of flunarizine on induced calcium transients as measured in fura-2-loaded neurons of the rat dorsal root ganglion. AB - The effect of the calcium entry blocker flunarizine on a high-potassium induced increase of intracellular free calcium was studied. The experiments were done with neurons isolated from rat dorsal root ganglia and loaded with the calcium sensitive dye fura-2. The increase of calcium induced by 60 mmol/l potassium was abolished after removal of extracellular calcium, was reversibly reduced by 50 mumol/l cadmium (76% inhibition), 50 mumol/l nickel (25% inhibition) and 10 mumol/l nifedipine (18% inhibition), and reversibly increased after removal of extracellular sodium (26% increase). The potassium induced increase of intracellular calcium is, therefore, mediated by transmembrane calcium influx, probably to a large extent through cadmium-sensitive calcium channels. Flunarizine (5 min incubation followed 1 min wash-out) reduced the amplitude of the high-potassium induced calcium increase in a dose-dependent manner (Kd = 370 +/- 100 nmol/l; mean +/- SEM; n = 8), causing complete inhibition at a concentration of 10 mumol/l in the majority of cells. Flunarizine (> or = 1 mumol/l) caused a reversible increase of the resting level of intracellular calcium in some cells, an effect which disappeared in the absence of extracellular calcium. The drug (1 mumol/l had no influence on the time course of recovery of intracellular calcium subsequent to a rise induced by high-potassium or by the calcium ionophore A23,187. It is concluded that flunarizine acts as an inhibitor of depolarization-mediated calcium influx. At a concentration of 1 mumol/l, the drug presumably has no effect on cellular calcium extrusion and/or sequestration mechanisms. PMID- 8232605 TI - Effects of hypoxia on stimulus-release coupling mechanisms in cultured bovine adrenal chromaffin cells. AB - To clarify the effects of hypoxia on stimulus-release coupling, we have examined the effects of hypoxia on nicotine-induced catecholamine (noradrenaline and adrenaline) release from, and 22Na+ influx, 45Ca2+ influx and cytosolic free Ca2+ concentration ([Ca2+]i) in, cultured bovine adrenal chromaffin cells. Experiments were carried out in media pre-equilibrated with 21% O2/79% N2 (control) or with 0% O2/100% N2 (hypoxia). Cells were stimulated with either nicotine (activating nicotinic acetylcholine (ACh) receptors) or a high K+ concentration (55 mmol/l KCl; directly activating voltage-dependent Ca2+ channels). Hypoxia reduced both nicotine- and high K(+)-induced catecholamine releases from the cells, but the reduction of the former (to about 30% of the control value) was more pronounced than that of the latter (to about 40% of the control value). Nicotine-induced 22Na+ influx, which is considered to reflect the function of nicotinic ACh receptors, was inhibited by hypoxia. Both nicotine- and high K(+)-induced 45Ca2+ influx into the cells were reduced by hypoxia, but the reduction of the former was more pronounced than that of the latter. Nicotine- and high K(+)-induced increases in [Ca2+]i were reduced by hypoxia to about 30% and 40% of the control values, respectively. These results suggest that hypoxia reduces cation influxes (Na+ and Ca2+) through both the ligand-gated cation channels of the nicotinic ACh receptor and the voltage-dependent Ca2+ channels. PMID- 8232606 TI - Pronounced direct inhibitory action mediated by adenosine A1 receptor and pertussis toxin-sensitive G protein on the ferret ventricular contraction. AB - An adenosine A1 receptor agonist R-N6-phenylisopropyladenosine (R-PIA) elicited a pronounced negative inotropic effect with the EC50 value of 0.69 mumol/l in the presence of a beta-adrenoceptor blocking agent bupranolol (0.3 mumol/l) in the isolated ferret papillary muscle. The negative inotropic effect of R-PIA was not associated with changes in cyclic AMP level. Adenosine and other A1 receptor agonists also elicited a negative inotropic effect. DPCPX (1,3-dipropyl-8 cyclopentyl xanthine) antagonized the negative inotropic effect of R-PIA in a competitive manner (pA2 value = 8.4). The inhibitory action of R-PIA was markedly attenuated in the ventricular muscle preparation isolated from ferrets pretreated with pertussis toxin that caused ADP-ribosylation of 39 kDa proteins in the membrane fraction. In the membrane fraction derived from the ferret ventricle, [3H]-DPCPX bound to a single binding site in a saturable and reversible manner with high affinity (Kd value = 1.21 +/- 0.41 nmol/l; Bmax = 12.8 +/- 3.02 fmol/mg protein; n = 7). The binding characteristics of [3H]-DPCPX in the rat ventricle (Kd value = 1.51 +/- 0.09 nmol/l; Bmax = 12.7 +/- 1.47 fmol/mg protein; n = 5) were similar to those in the ferret. On the other hand, the content of G(o), a major pertussis toxin-sensitive G protein in the ferret heart, was much higher in the ferret than in the rat ventricle. The present results indicate that adenosine receptors may play an important role in the inhibitory regulation of ventricular contractility in the ferret in contrast to other mammalian species. The signal transduction process subsequent to agonist binding to A1 receptors including the pertussis toxin-sensitive G protein and ion channels may be responsible for the unique inhibitory action of adenosine in this species. PMID- 8232607 TI - Inhibition of endothelial derived relaxing factor (EDRF) aggravates ischemic acute renal failure in anesthetized rats. AB - The relative importance of endothelial derived relaxing factor (EDRF)/nitric oxide (NO) in maintaining kidney function in normal condition and in acute renal failure (ARF) were evaluated in inactin anesthetized rats. ARF was induced by unilateral occlusion of the left renal artery (40 min) followed by reperfusion, with the contralateral kidney serving as normal control. This protocol resulted in marked reductions in renal plasma flow (RPF), glomerular filtration rate (GFR) and increases in fractional sodium excretion (FENa) and urinary protein excretion in the post-ischemic kidney in comparison to the contralateral normal kidney. Administration of the nitric oxide (NO) synthase inhibitor NG--monomethyl-L arginine (0.25 mg/kg per min, L-NMMA) exacerbated the ischemia-induced changes in renal functions as reflected by further reductions in urine flow (V), GFR, marked sodium wasting and renal edema. Pretreatment of the animals with NO precursor L arginine (2.5 mg/kg per min, L-Arg) abolished the detrimental effects of L-NMMA in ARF. In contrast, D-Arginine (2.5 mg/kg per min, D-Arg) failed to reverse the detrimental effects of L-NMMA. Infusion of L-Arg alone also resulted in improvements in RPF and GFR in the ischemic kidney. The results of the present study suggest that the function of the ischemic kidney is sustained by EDRF/NO and is thus more sensitive to NO synthase inhibition. PMID- 8232608 TI - Action of cisapride on rat colonic secretion. AB - The stimulating effect of cisapride on the motility of the digestive tract is well known. However, there are only a few studies on the influence of this drug on the absorptive or secretory activity of the colonic mucosa. In the present study, the ability of cisapride to alter the mural transport of water and electrolytes in the colon and its effects on mucus secretion and albumin permeation were studied. The effects of cisapride on the rat colon in vivo were studied under different conditions, by means of an instillation of sodium acetate solution at pH 6.9, which induced absorption of water and electrolytes, and in two models of colonic secretion, one employing the instillation of an acetic acid solution at pH 2.9 and the other, an intravenous infusion of 5-hydroxytryptamine (serotonin) 45 micrograms.kg-1.min-1 together with intracolonic instillation of sodium acetate. Cisapride (i.v.), at a dose of 0.32 mg.kg-1, in rats whose colon was instilled with sodium acetate (pH 6.9) induced an increase in sodium absorption and a reduction in water absorption. Cisapride (i.v.), at doses of 0.32, 0.64 and 1.0 mg.kg-1, inhibited the secretion of water, Na+, Cl-, and mucus and the permeation of albumin induced by acetic acid instillation or by serotonin infusion. It is concluded that the effect of cisapride on the colonic mucosa varies in accordance with the functional mucosal conditions and that this action may be of clinical importance. PMID- 8232609 TI - Role of Na+/Ca++ exchange in the relaxant effect of sodium taurocholate on the guinea-pig ileum smooth muscle. AB - Sodium taurocholate (NaTC), at concentrations below the critical micellar concentration, caused a transient relaxation of isolated guinea-pig ileum smooth muscle strips. The relaxation was not inhibited by previous incubation with either 10 microM ouabain, 0.4 mM d-tubocurarine or 0.5 microM apamin, ruling out the participation of hyperpolarization of the plasma membrane induced by either stimulation of Na+/K+ ATPase or by opening of Ca(++)-dependent K+ channels. In guinea-pig ileum smooth muscle cultured cells, addition of NaTC (1 mM) stimulated Na+ uptake and Ca++ efflux. The relaxation induced by NaTC was inhibited by 3',4' dichlorobenzamil, a blocker of the Na+/Ca++ exchanger. Preincubation with NaTC, or its addition during the early stage of the tonic response of the ileum to acetylcholine, enhanced that response, whereas a relaxation was observed when NaTC was added at the late stage of the acetylcholine response. In cultured cells, NaTC potentiated the stimulation of Ca2+ influx by acetylcholine. Our results suggest that NaTC acts on the smooth muscle cell membrane causing a stimulation of the Na+/Ca++ exchange mechanism. PMID- 8232610 TI - Identification of P450 enzymes involved in metabolism of verapamil in humans. AB - The calcium channel blocker verapamil[2,8-bis-(3,4-dimethoxyphenyl)-6-methyl-2 isopropyl-6- azaoctanitrile] is widely used in the treatment of hypertension, angina pectoris and cardiac arrhythmias. The drug undergoes extensive and variable hepatic metabolism in man with the major metabolic steps comprising formation of D-617 [2-(3,4-dimethoxyphenyl)-5-methylamino-2 isopropylvaleronitrile] and norverapamil [2,8-bis-(3,4-dimethoxyphenyl)-2 isopropyl-6-azaoctanitrile]. The enzymes involved in metabolism of verapamil have not been characterized so far. Identification of these enzymes would enable estimation of both interindividual variability in verapamil metabolism introduced by the respective pathway and potential for metabolic interactions. We therefore characterized the enzymes involved in formation of D-617 and norverapamil. The maximum rate of formation of D-617 and norverapamil was determined in the microsomal fraction of 21 human livers which had been previously characterized for the individual expression of various P450 enzymes (CYP1A2, CYP2C, CYP2D6, CYP2E1 and CYP3A3/4) by means of Western blotting. Specific antibodies directed against CYP3A were used to inhibit formation of D-617 and norverapamil. Finally, formation of both metabolites was investigated in microsomes obtained from yeast cells which were genetically engineered for stable expression of human P450. Formation of D-617 was correlated with the expression of CYP3A (r = 0.85; P < 0.001) and CYP1A2 (r = 0.57; P < 0.01) in the microsomal fraction of 21 human livers after incubation with racemic verapamil.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232611 TI - A comment about physician-assisted suicide. PMID- 8232612 TI - Doctors vs lawyers. PMID- 8232613 TI - Screening of asymptomatic patients. PMID- 8232614 TI - The bioethics resource group: a model for community bioethics. PMID- 8232615 TI - Screening for ovarian cancer: not worthwhile for most patients. PMID- 8232616 TI - Pulmonary surfactant. A historical perspective of how it came to be used in the treatment of respiratory distress syndrome in the neonate. PMID- 8232617 TI - Dorma con pesce. Scombroid icthyotoxicosis. PMID- 8232618 TI - Conflicts of interest in physician ownership of medical facilities. A clarification of present AMA and North Carolina Medical Society policy. PMID- 8232620 TI - Our country's mania for medical care is separating us from our constitutional heritage. PMID- 8232619 TI - Sex addiction. PMID- 8232622 TI - The return of the generalist physician. Initiatives of the University of North Carolina School of Medicine. PMID- 8232621 TI - An affordable package of medical care for all Americans is a higher goal than absolute freedom for physicians. A reply to Dr. Weaver. PMID- 8232623 TI - My time as a minority student at Meharry Medical College. PMID- 8232624 TI - To encourage or not to encourage our children to study medicine. PMID- 8232625 TI - Nurses plan to open dialysis facility in Philadelphia. PMID- 8232626 TI - AOPO sheds light on organ acquisition costs, responds to survey data. PMID- 8232627 TI - Exceptions will help nephrologists deal with new self-referral rules. PMID- 8232628 TI - Can a global payment system work for the ESRD program? PMID- 8232629 TI - When dialysis only prolongs the dying process. PMID- 8232630 TI - Dialysis should be withdrawn when the burdens outweigh the benefits. PMID- 8232631 TI - Dialytic care is an unprecedented privilege. PMID- 8232632 TI - ESRD program at the crossroads of health care reform. PMID- 8232633 TI - HCFA data show CCPD use increased by 29.2% in 1992. PMID- 8232634 TI - There is no job security in a failed organization ... lessons for labor and management. PMID- 8232635 TI - An interview with Marty Minniti. Interview by Connie R. Curran. AB - The SNI Companies are nurse-owned, nurse-managed organizations founded on the concept that individual independence and autonomy promotes professional excellence and the ability to achieve personal and professional goals. Marty Minniti, BS, RN, CCRN, founder, president, and chief executive officer, discusses SNI Companies, and the rewards and challenges of entrepreneurial and intrapreneurial nursing. PMID- 8232636 TI - Nursing: an important component of hospital survival under a reformed health care system. AB - As health care moves into a competitive environment, hospitals will be pressed to create and maintain a competitive edge to survive. Core clinical staff, such as RNs, are a key component in increasing quality, understanding patient care needs, and balancing costs in a reformed health care system. PMID- 8232637 TI - Health reform--its potential impact on hospital nursing service. AB - The passage of major health reform measures in the United States focusing on universal access and cost containment could potentially have a significant impact on the delivery of nursing services in our nation's acute-care facilities. If President Clinton's global budgetary target concept, which is used in most western countries to constrain expenditures, is implemented in the United States, the experience of Canadian medium-size and teaching hospitals suggests that such a cost-containment approach would have a modest impact on nurse staffing patterns here. PMID- 8232638 TI - Shared governance and the nursing practice environment. AB - An evaluation of the impact of shared governance on staff nurse perceptions of management style, group cohesion, job stress, job satisfaction, and anticipated turnover provides support for claims that shared governance improves the nursing practice environment. PMID- 8232639 TI - Work restructuring: the process of decision making. AB - Work restructuring at a 269-bed hospital resulted in a 20% reduction in the nursing operations budget. A complex process, work restructuring must be logical and requires both interdisciplinary workers and committed leadership. PMID- 8232640 TI - Analysis of state and local hospital employed RN labor markets. AB - Analysis and knowledge of state and local RN labor markets can facilitate nurse retention and recruitment by allowing nurse administrators to better predict market changes and maintain a stable RN workforce. PMID- 8232641 TI - Point of care technology: impact on people and paperwork. AB - A systematic evaluation of a bedside computer system for nursing documentation indicates that point of care technology has a positive direct impact on the efficiency, effectiveness, and satisfaction of the nursing staff, and a positive indirect influence on other members of the health care team in their delivery of patient care. PMID- 8232643 TI - The nurse manager and service-line management. PMID- 8232642 TI - Read this. Now! PMID- 8232644 TI - Evaluating the effectiveness of an intravenous therapy team. PMID- 8232645 TI - Congressional debate focuses on budget reconciliation, not health care reform. PMID- 8232646 TI - An ivory tower or a velvet rut? PMID- 8232647 TI - An interview with Kristine Gebbie. Interview by Carmella A. Bocchino. AB - The National Commission on AIDS ended its 4 years of operation just as President Clinton named the Administration's first AIDS Policy Coordinator, Kristine Gebbie, MN, RN, FAAN. "A strong, consistent voice of leadership" was a key recommendation from the commission and Ms. Gebbie promises to be the stimulus for such leadership in both the public and private sectors. As the number of new AIDS cases continues to increase, hospitals and other care providers will be burdened with the financial challenge of caring for these patients, many of them uninsured. Calling upon her strong public health background, Ms. Gebbie discusses some of her goals for this new position with Nursing Economic$. PMID- 8232648 TI - Building TQM into nursing management. AB - Total quality management (TQM) is a management philosophy that addresses problems currently faced by health care, specifically reducing costs while improving quality of services. As hospital administrators embrace this new management style, nurse executives and managers will be challenged to implement TQM. Building TQM into nursing management will improve quality and reduce costs while meeting the needs of health care customers. PMID- 8232649 TI - Health care corporatization and the employment conditions of nurses. AB - There has been little study of the impact of the recent transition of ownership of hospitals and nursing homes, from voluntary and public to corporate, on the employment of nursing staffs. Non-state or non-federally owned hospitals and nursing homes in Florida were examined with regard to salaries, benefits, career advancement opportunities, and vacancy and turnover rates of the nursing staff. While salary levels were influenced primarily by local wage markets, significant differences were found in the benefit packages, career opportunities, staffing ratios, and vacancy and turnover rates among for-profit, not-for-profit, and publicly-owned facilities. Nurse administrators of corporate hospitals and nursing homes can lower the costs of high turnover rates in their nursing staffs by exploring improvements in benefit plans and career opportunities offered to their employees. PMID- 8232650 TI - A cost analysis of a professional practice model for nursing. AB - Costs of nursing care among hospital units that adopted a professional practice model (PPM) were compared with traditional nursing units. PPM inpatient units used fewer temporary personnel and nursing aides, resulting in similar RN costs but lower total nursing costs. However, PPM operating rooms were more costly, mostly because of the intense use of RNs. PMID- 8232651 TI - Redefining the purpose of patient classification. AB - One hospital's nursing staff used a structured process to challenge the purpose and function of the patient classification system. The result was internal development of a more effective system with significant cost savings. PMID- 8232652 TI - A study of the public's knowledge regarding advance directives. AB - Advance directives are legal documents used to indicate individuals' wishes concerning medical care and treatment in the event that they are no longer able to do so. This study assessed the level of knowledge of 157 adults concerning advance directives and other medical terminology related to end-of-life events. Results indicated that 57% to 79% of respondents demonstrated an understanding of four to five major elements related to advance directives. There were no significant differences in level of understanding among groups of differing ethnicity, religion, and gender. Nor were there significant correlations found between level of understanding and the respondents' age and years of education. PMID- 8232653 TI - RN students: unobtrusive cost-effective change agents. AB - A teaching learning project that empowers registered nurse (RN) to bachelor of science in nursing (BSN) senior nursing students to undertake cost containment and quality improvement pilot projects in the institutions where they work enables nursing education and nursing service to collaborate in promoting cost containment and quality improvement. Understanding how this process was implemented in one institution, as well as the outcomes, can help guide the application of this creative teaching strategy. PMID- 8232654 TI - Congressional attention finally turns to health care reform. PMID- 8232655 TI - Case management/outcomes management: the role of the nurse manager. PMID- 8232656 TI - [A nurse with a hernia nuclei pulposi]. PMID- 8232657 TI - [Standards for family practitioner's activities: a good beginning but no end]. PMID- 8232658 TI - [The importance of publication bias in medical-scientific literature]. PMID- 8232659 TI - [Cell and disease. VII. Cell-cell and cell-matrix interactions in pathological processes]. PMID- 8232660 TI - [Guidelines of the Dutch College of Family Practitioners]. AB - The Dutch College of General Practitioners (NHG) produces scientifically founded guidelines for general practice. They mark the emancipation of General Practice into a specialism in its own right and are used for education and quality control. They differ from national and international consensus guidelines, in that they are developed by general practitioners only, and that notice is taken of the specific predictive value of signs and tests in the setting of general practice. Moreover they usually go into more detail. NHG Standards are provided with additional Teaching Packages for use in comprehensive programme of continuous medical education. So far, 40 Standards have been published. They were very well received by general practitioners. Diffusion to the rest of the medical profession is under way. PMID- 8232661 TI - [Performance of family practitioners according to the guideline otitis media acuta of the Dutch College of Family Practitioners]. AB - OBJECTIVE: To establish to which degree the guidelines of the Otitis media acuta standard of the Netherlands college of general practitioners (NHG) are followed in practice and what is GPs' opinion of them. DESIGN: Survey. SETTING: 65 general practitioners settled throughout the Netherlands. METHOD: The GPs were recruited for the study by means of an advertisement and a personal approach. Opinions and performances were inventoried using a questionnaire and self-completion forms for successive doctor-patient contacts, respectively. RESULTS: Virtually all participants were familiar with the NHG standard and most were familiar with three central guidelines. Of these three, the recommendation of an active follow up policy for children up to the age of 2 was known to the fewest GPs, and this recommendation also received the least support. In the 327 consultations recorded there was good agreement between actual performance and the guidelines, especially as regards history-taking, physical examination and diagnosing. Regarding management and patient information the agreement was less. Concerning the follow-up policy for young children and the prescribing of antibiotics, actual performance clearly differed from the NHG standard, possibly because the relevant guidelines were less well known. CONCLUSION: The results of this study are relevant to the planning of GP education and the evaluation of the guidelines themselves. PMID- 8232662 TI - [Estimation of the possibility of HIV or hepatitis B infection transmission in ambulatory care of substance dependence]. AB - OBJECTIVE: To estimate the risk of HIV or hepatitis B infection after pricking accidents in the ambulant care and treatment of drug addicts. DESIGN: Risk estimation. SETTING: Aid station for drug addicts in Nijmegen. METHOD: Inventorization of the pricking accidents at the above-named aid station and calculation of the risk of infection. RESULTS: During 30 years on the job, a worker in addict care outside Amsterdam runs a 0.007% risk of job-related HIV infection and a 4.5% risk of hepatitis B infection. The corresponding figures for Amsterdam are 0.05% and 7%, respectively. The job-related risk for workers in general hospitals in Amsterdam is 0.0012-0.015% for a job-related HIV infection and 1-5% for a job-related hepatitis B infection. CONCLUSION: The risk of workers in an aid station for drug addicts of contracting a job-related HIV or hepatitis B infection is relatively high compared with workers in health care in general. An active infection prevention policy for workers in addict care is to be recommended. PMID- 8232663 TI - [Ovum donation: is the shortage past? Ethical considerations]. PMID- 8232664 TI - President's theme. PMID- 8232665 TI - 1962-1992: thirty years of changes at the University of Nebraska College of Medicine. PMID- 8232666 TI - Bone marrow transplantation at the University of Nebraska Medical Center. AB - The bone marrow transplant program at UNMC is currently one of the most active programs in the country. The benefits to patients who are cured of disease by transplantation cannot be measured. The large volume of clinical and basic science research related to transplantation has enhanced the academic stature of UNMC. The combination of patient care, education, clinical research, and basic science research provides an excellent model for the operation of an academic medical institution. PMID- 8232667 TI - Adverse drug reactions in the elderly: use of a community based E-coding data base. PMID- 8232668 TI - The diagnosis of urinary tract infections during pregnancy. PMID- 8232669 TI - Managed care and the Nebraska Medical Association. PMID- 8232670 TI - Ileal pouch-anal anastomosis: long term outcome and patient satisfaction. PMID- 8232671 TI - Psychiatrist practice patterns: a comparison of Omaha and Lincoln. AB - Blue Cross and Blue Shield of Nebraska physician claims data were used to generate and test the hypothesis that there are major differences in psychiatrist practice patterns between the Omaha and Lincoln areas, as evidenced by billing practices. There were extreme differences between the population-based claim rates for the Omaha and Lincoln areas which led to the hypotheses stated above. Provider-based methodology was used to demonstrate that, for Nebraska psychiatrists, as patient load increases, number of charges per patient decreases. Psychiatrists with large market shares for the two respective areas were compared also. Fifty percent of the Omaha psychiatrists with large market shares had a significantly high number of claims per patient, whereas only 25% of the Lincoln psychiatrists (with large market shares) had significantly high numbers of claims per patient. Our hypothesis that there are major differences between Omaha and Lincoln psychiatrist practice patterns was supported. Recommendations include follow-up with the "Patient Satisfaction Survey", continued monitoring, and investigation by the medical community to determine causes for the wide discrepancies in psychiatrist practice patterns. PMID- 8232672 TI - Automobile air bag eye injuries. AB - Until improvements in the design of air bags have minimized the risk of serious ocular trauma, practitioners must be alert to the possibility of severe eye damage any time the device is activated. Prompt ophthalmological referral and prompt treatment of any ocular injury can result in a lower incidence of permanent visual loss. PMID- 8232673 TI - Child abuse revisited--the X-ray differential. PMID- 8232674 TI - First trimester antepartum obstetrical ultrasound examination. PMID- 8232675 TI - [Therapy of metabolic myopathies]. AB - Metabolic myopathies are subdivided into disturbances of anaerobic cytoplasmic and aerobic mitochondrial metabolism. With the exception of carnitine deficiency these myopathies are based on enzymopathies. Since gene therapy is not yet available no causal therapy is possible. This paper discusses possibilities for symptomatic therapy. Good results are found with carnitine substitution. Enzymopathies can be improved by using other metabolic pathways or by addition of co-factors of the impaired pathways. This leads to a reduction of myalgia, cramps, and endurance exercise intolerance. PMID- 8232676 TI - [Interstitial radiosurgery of low grade glioma]. AB - In this retrospective study the outcome of 324 patients with low-grade gliomas (WHO-Grade II) is evaluated, who from 1979 to 1991 underwent stereotactic biopsy and interstitial radiosurgery (Iodine-125). The patients were selected for interstitial radiosurgery if the tumor was unifocal, well-circumscribed and in diameter < or = 4 cm. Of these patients 251 had astrocytomas, 29 had oligodendrogliomas, and 44 had oligoastrocytomas. All patients presented with progressive neurological deficit or showed evidence of CT/MRI confirmed tumor growth. The preoperative performance status was > or = 70%. Diffusely infiltrative, non-delineated gliomas and gliomas crossing the midline were excluded. Five-year survival rates were 65% for astrocytomas, 80% for oligoastrocytomas and 58% for oligodendrogliomas. In the multivariate analysis age was a significant factor predicting survival. Patients who received temporary implants had a significantly better outcome than those with permanent implants. Radiation toxicity (3.1%) was seen mostly among the permanent implants. No patients required reoperation due to radionecrosis. The findings in this subgroup of gliomas indicate that interstitial radiosurgery using Iodine-125 is effective in controlling tumor growth and is well tolerated. Patients with differentiated and circumscribed gliomas particularly benefit from the treatment. PMID- 8232677 TI - [Coping with illness in myasthenia gravis]. AB - Within the past few years, the introduction of immunosuppressants in the treatment of myasthenia gravis, has turned this severe, chronic and life threatening disease into a moderate illness with good chances of control or even remission. Of 44 patients whose coping behavior and course of disease we investigated, only seven experienced a significant change in severity of the disease within nine months. This consistency in the course of the disease was also reflected in the forms of coping with the disease: in contrast to the situation two decades ago, today's myasthenia gravis patients are no longer caught up in the dilemma between passive dependency and active resistance, most of them adopting an attitude of calm acceptance. According to the Berne Forms of Coping (BEFO) which we applied, this attitude is expressed as a pattern of passive cooperation, acceptance, distraction and relativization. This coping pattern remained largely intact even in relation to the severity of the disease and the retrospectively assessed course of the disease. We present two cases to illustrate the influence of coping behavior on the course of the disease. Comparison with two other patient groups (rheumatoid arthritis and hip osteo arthrosis) showed that there is no disease specificity of coping behavior. It can be assumed that there is a basic pattern in coping with chronic diseases, the manifestation of which is dependent on the severity and prognosis of the disease. If depressive reactions to the disease are excluded, the prevalence of longer term pre-existent psychiatric disorders among MG patients corresponds to the average for the general population, although there is a relatively high incidence of anxiety disorders.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232678 TI - [Heparin-induced extracorporeal LDL precipitation (HELP). A new therapeutic possibility in cerebral multi-infarct dementia]. AB - High fibrinogen levels have been assessed in cerebrovascular disease with a direct relationship to blood rheology. Heparin-induced extracorporeal LDL precipitation (HELP) is a method that effectively reduces fibrinogen and lipoproteins at the same time, thus improving the hemorheologic pattern. We studied 45 patients with cerebral multiinfarct dementia. After computer randomization 30 patients received a HELP treatment, while the remaining 15 patients served as controls. HELP produced an immediate and statistically significant reduction on parameters relevant to hemorheology, such as fibrinogen (P < 0.001), total cholesterol, low-density lipoprotein, triglycerides (P < 0.0001 each) and whole blood viscosity (at high and low shear rate) as well as plasma viscosity and red cell transit time (P < 0.01 each). The treated group showed improvement after HELP (P < 0.05 for each test) on the Mathew Neurological Scale, Mini Mental State Examination and Activities-of-Daily-Living Test and also relative to the untreated controls (P < 0.05 each). These results may be regarded as a basis for some reflections about new hemorheological interventions in cases of cerebral multiinfarct dementia. PMID- 8232679 TI - [Munich Intelligibility Profile. Studies of reliability and validity]. AB - The 'Munchner Verstandlichkeitsprofil (MVP)' (Munich Intelligibility Profile) is a PC-based method for assessment of the intelligibility of dysarthric speakers. Analyses of its reliability and validity were based on a total of 101 patients and 16 normal speakers as well as a group of 45 listeners. The reliability of the overall MVP score was determined using non-parametric analyses of variance and between-listener correlations. Retest reliability was measured for a subgroup of 5 listeners. Test validity was analyzed by correlating MVP scores with three external criteria (expert ratings, listeners' ratings, transcripts). Further, relevant aspects of the test construction were tested empirically. Finally, regression analyses were carried out in order to demonstrate that different MVP lists are equivalent in predicting transcription-based intelligibility scores. PMID- 8232680 TI - [Clinical course of paraneoplastic limbic encephalitis]. AB - A 49-year-old woman presented with increasing memory loss without dementia. The EEG showed slow activity over the temporal lobe. MRT revealed temporal areas of increased signal intensity without gadolinium enhancement. The diagnosis of limbic encephalitis was made after detection of a bronchial carcinoma. A MRT control examination after chemotherapy showed resolution of the abnormalities. This observation may indicate that chemotherapy has modified that part of the immunological system responsible for induction of limbic encephalitis. PMID- 8232681 TI - [Apraxia in eyelid opening after stereotactic sub-thalmotomy. Case report and review of the literature]. PMID- 8232682 TI - [Oculomotor paralysis as the leading symptom of meningovascular syphilis. Report of two patients and review of the literature]. AB - A brain stem infarction in the area of the nerve fascicle of the oculomotor nerve could be demonstrated by NMR to be the cause of a complete oculomotor nerve palsy in a patient with neurosyphilis. A second case presented the very rare finding of an isolated bilateral incomplete oculomotor nerve paresis with external paresis on the right and internal paresis on the left side. PMID- 8232683 TI - [Neurogenic intermittent claudication of the femoral nerve caused by occlusion of the common iliac artery]. AB - A 52 year old heavy smoker complained of paresthesiae and pain at the ventral side of the right thigh and the antero-medial side of the right lower leg as well as weakness of the right quadriceps femoris during exercise. Clinical examination revealed a paresis of the right quadriceps, hypesthesia and hypalgesia in the area of the femoral nerve and a reduced right patellar reflex after 10 min walking. An occlusion of the right common iliac artery was diagnosed by angiography. Following transluminal angioplasty and implantation of an intravascular stent, the patient was free of symptoms. On the basis of the clinical observations following recanalisation of the common iliac artery, the symptoms can best be explained by a reduced perfusion of the iliolumbar artery supplying the upper part of the femoral nerve, causing ischemia of the femoral nerve during exercise. In conclusion, stenosis/occlusion of the common iliac artery should be considered as a differential diagnosis of quadriceps weakness and paresthesia in the area of the femoral nerve associated with exercise. PMID- 8232684 TI - [Endogenous bilateral compression syndrome of the suprascapular nerve. Overview and case report]. AB - Entrapment of the suprascapular nerve with shoulder pain and atrophic paresis of both the supraspinatus and infraspinatus muscles has been known since the fifties. So far a bilateral entrapment of the suprascapular nerve has rarely been described. We present a report of an endogenous bilateral entrapment neuropathy of the suprascapular nerve and discuss the differential diagnoses in relation to therapeutic and prognostic consequences. PMID- 8232685 TI - [Course of bacterial infections in high-dose therapy with clozapine]. AB - The influence of neuroleptic drugs on body temperature regulation is well established. Even with atypical neuroleptic drugs such as clozapine, hyperthermia has been observed as well as the malignant neuroleptic syndrome. It is likely that the influence of clozapine affects the immune response. A case is reported in which afebrile angina tonsillaris and endocarditis developed, as a result of high-dosage therapy with clozapine over several years. Besides the special features of the course, differential diagnosis, therapy and possible influence on the body temperature and the immune response are discussed. PMID- 8232686 TI - [Is neopterin a diagnostic marker of early neurologic HIV manifestation?]. PMID- 8232687 TI - Dutch Society of Gastroenterology and Dutch Society of Hepatology. 18-19 March 1993 . Meeting abstracts. PMID- 8232688 TI - Hyponatraemia: SIADH or SIAD? PMID- 8232689 TI - The relationship between subjective vasospastic complaints and finger blood flow measurements in Raynaud's phenomenon. AB - BACKGROUND: Objective circulatory tests in patients with Raynaud's phenomenon have only rarely been related to subjective complaints. METHODS: In 55 Raynaud patients the correlation between daily self-recorded frequency, duration and severity of vasospastic attacks and the measurement of finger skin blood flow during local cooling was investigated. RESULTS: No significant correlations were found between complaints as registered in the diaries and finger skin temperature and laser Doppler flux before, during and after a standardized local cooling procedure. CONCLUSION: Finger skin blood flow measurements do not necessarily reflect digital vasospasm in daily life. Besides studies on the diagnostic value and reproducibility of skin blood flow measurements in Raynaud's phenomenon, the relationship of test results to subjective complaints should be established. PMID- 8232690 TI - Human immunodeficiency virus, fever, dyspnoea and a dry cough. Expect the unexpected? AB - This report describes an HIV-seropositive patient with symptoms suggestive of Pneumocystis carinii pneumonia. The final diagnosis of pulmonary embolism was delayed because initially only HIV-specific complications were considered. The relation between AIDS and pulmonary embolism is discussed briefly. PMID- 8232691 TI - Carcinoma of the papilla of Vater in a brother and sister with familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) is characterized by the development of adenomas throughout the entire gastrointestinal tract. We describe a brother and sister with FAP who developed adenocarcinoma around the papilla of Vater. Incidence, premalignancy, treatment and the need for prophylactic endoscopy are discussed. PMID- 8232692 TI - Wilson disease. PMID- 8232693 TI - Thoughts on the thymus and the T-lymphocyte repertoire. Relevance to the tolerance of the immune response. AB - The thymus is a central lymphoid organ, and has one important function in the immune system. It generates immunocompetent T (= thymus-dependent) cells. 'Immunocompetence' is the single most important characteristic acquired during the differentiation process of T-cells, from progenitor cell in the bone marrow to mature cell in peripheral lymphoid organs and blood. The basis of competence is recognition, mediated by the T-cell receptor on the cell surface. Using this receptor, T-cells recognize antigens only in the form of antigenic fragments complexed with molecules of the major histocompatibility complex. Within the thymus, progenitor cells rearrange gene segments encoding the various parts of the receptor molecule. After expression of all possible receptors (the potentially available repertoire) selection processes occur, yielding only those T-cells that have a receptor by which antigens can be recognized together with molecules of the major histocompatibility complex, and that do not recognize autoantigens (the actual repertoire). This process of selection is considered as a unique function of the thymus. In this review, first some histophysiological aspects of the thymus are described. Next, the events that occur during the sojourn of the lymphocytes in the thymus are presented. Finally, these data are considered with regard to thymic abnormalities. These include acute and age associated involution, and the generation of autoimmune reactivity due to disturbed intrathymic selection. PMID- 8232694 TI - 1-Desamino-8-D-arginine vasopressin (DDAVP) in patients with congenital nephrogenic diabetes insipidus. AB - In healthy subjects, intravenous infusion of the selective V2-vasopressin receptor agonist 1-desamino-8-D-arginine vasopressin (DDAVP, 400 ng/kg in 10 min) causes a marked increase in heart rate with a slight decrease in diastolic blood pressure. These haemodynamic responses are associated with increments in the plasma levels of renin, noradrenaline (NA), clotting factor VIII (FVIII:C), von Willebrand factor (vWF:ag), and tissue-type plasminogen activator (t-PA), and a fall in the plasma level of plasminogen activator inhibitor (PAI). None of these changes was observed in 3 patients with congenital nephrogenic diabetes insipidus (NDI), who had a genetic defect of the V2-receptor. Plasma AVP levels in these patients were normal or slightly elevated, which makes it unlikely that the lack of DDAVP responsiveness was caused by down-regulation of vasopressin V1 receptors. In one NDI patient, arginine vasopressin (AVP) was given in incremental doses (62.5-4000 pg/kg/min). The heart rate and blood pressure responses to AVP were normal, indicating the absence of a V1-receptor defect. The responses of vWF:ag and t-PA to venous occlusion in the patients with NDI were similar to those in 5 healthy volunteers, which indicates that in NDI the endothelial release of both vWF:ag and t-PA is normal. We conclude that DDAVP causes its effects on heart rate and blood pressure, and on the plasma levels of renin, noradrenaline, FVIII:C, vWF:ag, and t-PA through V2-receptor stimulation. PMID- 8232695 TI - Quantitative assessment of pancreatic function by magnetic resonance in cystic fibrosis. AB - To determine the value of magnetic resonance imaging in the analysis of pancreatic function in cystic fibrosis, 11 adult cystic fibrosis patients with exocrine pancreatic insufficiency were studied by magnetic resonance and a set of non-invasive tests of exocrine and endocrine pancreatic function. Magnetic resonance imaging, comprising the calculation of intensity ratios (pancreas/liver) of the pancreas on T1 ([SE] TR/TE = 300/20) and T2 weighted ([SE] TR/TE = 2000/50-100)images and the measurement of several dimensions of the pancreas, was compared with the results of the indirect pancreatic function tests. Although magnetic resonance showed the typical morphology of the insufficient pancreas in cystic fibrosis, including atrophy, lobular fatty enlargement and infiltration, it was not related to the quantitative exocrine and endocrine function assessed by indirect pancreatic function tests. PMID- 8232696 TI - The assessment of the functional capacity of the hypothalamic-pituitary-adrenal axis by measurement of basal plasma and urinary cortisol in comparison with insulin-induced hypoglycaemia and metyrapone tests. AB - In 58 patients with a pituitary adenoma or hypothalamic-pituitary disease an insulin-induced hypoglycaemia test and a metyrapone test were performed. The results of these tests were compared with morning plasma cortisol levels and daily urinary cortisol excretion as indicators of insufficiency of the pituitary adrenal axis. Basal unstressed urinary cortisol excretion was insufficient in 20 cases. These patients, needing life-long glucocorticoid substitution therapy, were excellently detected by both tests and daily urinary cortisol excretion. The predictive value of the morning plasma cortisol level was inferior to these. Five cases with sufficient basal cortisol excretion showed a defective adrenal response to hypoglycaemia. These patients were not discriminated by the metyrapone test, urinary cortisol excretion or plasma cortisol levels. It is concluded that urinary cortisol excretion can safely replace the hypoglycaemia and metyrapone test for the detection of insufficient basal cortisol production in patients with hypothalamic and/or pituitary disorders. PMID- 8232697 TI - Cortical blindness due to cyclosporin. AB - We here report the occurrence of cortical blindness in a kidney transplant recipient treated with cyclosporin. The cortical blindness was the result of cyclosporin toxicity and resolved spontaneously after cyclosporin dose reduction. PMID- 8232698 TI - Methimazole-induced agranulocytosis preceded by transient granulocytopenia. A case report. AB - A 36-year-old patient, euthyroid under methimazole treatment, was admitted because of an active Graves' ophthalmopathy and found to have a transient granulocytopenia. Forty-five days after this episode she developed classical agranulocytosis because of which the methimazole was stopped. The agranulocytosis occurred more than 20 months after the initiation of antithyroid drug therapy. Other causes for the initial phase of granulocytopenia were not found. PMID- 8232699 TI - Long-term application of human polyclonal hepatitis-B immunoglobulin to prevent hepatic allograft infection. A review of the literature and presentation of five cases. AB - Liver transplantation has evolved as the therapy of choice in end-stage liver disease. However, liver transplantation for viral liver disease is associated with recurrence of viral hepatitis in the graft. In this paper we discuss the mechanisms of liver cell damage, the clinical impact of recurrent infection and the use of hepatitis-B-specific immunoglobulin (HBIg) to prevent recurrent infection in the allograft. In addition, we propose an HBIg schedule tailored to each individual patient. Based on the excellent survival results published for acute hepatic failure and chronic liver failure with latent hepatitis B virus (HBV) or hepatitis delta infection, we think that no contraindication exists for liver transplantation in these cases. For patients with active viral replication, liver transplantation should only be considered within a scientific frame to prevent reinfection of the allograft. PMID- 8232700 TI - The role of modification of lipoproteins and of the immune system in early atherogenesis. AB - Not only the plasma cholesterol level, but also postsecretory modifications of lipoproteins appear to be of influence in atherogenesis. Evidence that several forms of modification, especially oxidation, occur in vivo is rapidly accumulating, although their clinical relevance remains uncertain. Modification of lipoproteins has been demonstrated in persons with such well-known risk factors of premature atherosclerosis as smoking, diabetes mellitus and hyperlipidaemia. Because there is a relation between the amount of natural antioxidants in the plasma and the risk of atherosclerosis, and because exogenous antioxidants appear to retard atherosclerosis without influencing the plasma cholesterol level, antioxidants may prove to be of use in the prevention of atherosclerosis. There are strong indications for a role of the immune system in atherogenesis. Modified lipoproteins are highly immunogenic and stimulate immunocompetent cells to secrete vasoactive factors and cytokines. From animal studies it appears that pro- and antioxidative conditions can modulate these processes. It is concluded that additional research on the relation between lipoprotein modification and the immune system, and on the possible beneficial effects of antioxidants in atherogenesis is warranted, not only to elucidate further the mechanism of atherosclerosis, but also to develop new approaches to the prevention of atherosclerosis. PMID- 8232701 TI - Phenprocoumon-induced liver failure. PMID- 8232702 TI - No acute toxicity after massive anticoagulant overdose. PMID- 8232703 TI - A randomized controlled trial on the treatment for acute partial ischemic stroke with acupuncture. AB - The effectiveness of acupuncture in acute stroke remains largely untested and unproved. A randomized, controlled trial was carried out to study the feasibility of acupuncture in combination with conventional supportive treatment for acute stroke patients. A total of 30 patients, aged 46-74, with the onset of symptoms within 36 h were enrolled into the study after appropriate screening. All patients gave informed consent. Basing on the same supportive treatment, patients were randomly assigned to a treatment with or without acupuncture. The procedure and acupoint selection were discussed and decided through several meetings of a group of senior acupuncture doctors in Taiwan. Acupuncture was applied 3 times/week for 4 weeks. During the study period, there were no problems in conducting this trial in terms of patient availability and acceptance, and physician cooperation. A significantly better neurologic outcome was observed in the acupuncture group on day 28 and day 90. The improvement in neurologic status was greatest in patients with a poor neurologic score at baseline. There were no important side effects except for one episode of dizziness related to acupuncture treatment. The data and results of this study will be used as a guideline for planning a full-scale clinical trial, e.g. sample size calculation, method of randomization with stratification of prognostic factors, choosing acupuncture points and technique of acupuncture. PMID- 8232704 TI - Induction of cytochrome P-450 enzymes via tobacco smoke: a potential mechanism for developing resistance to environmental toxins as related to parkinsonism and other neurologic diseases. AB - Case-control studies have consistently demonstrated a reduced risk of Parkinsonism in smokers. A potential mechanism of protection is based on the 'induction' of cytochrome P-450 enzymes by polyaromatic hydrocarbons in tobacco smoke. Cytochrome P-450 enzymes are responsible for biomodification and detoxification of foreign substances. Stimulation of these enzymes in the olfactory epithelium and in the liver may lead to increased metabolism of xenobiotics, thereby preventing environmental neurotoxins from reaching target organs. Polymorphic expression of this family of isozymes may confer a unique susceptibility to some neurologic diseases. PMID- 8232705 TI - Infections in childhood and adolescence in multiple sclerosis. A case-control study. AB - A case-control study on multiple sclerosis was conducted in Western Norway during the years 1986-1988. Included were 155 persons with multiple sclerosis and 200 controls, marginally matched according to age, sex and area of residence. The mean age at measles infection was for the cases 6.6 years and for the controls 5.7 years (p = 0.06). The cases had more frequently experienced bronchitis and/or pneumonia in the age group 11-15 years (OR = 3.20, 95% confidence interval 0.96 10.63). Tonsillectomies were reported more frequently by the cases. The odds ratio was especially high for those treated at age 0-6 years (OR = 3.44, 95% confidence interval 1.63-7.27). The results are consistent with the idea of MS as an age-dependent, host-immune response to infection during childhood or adolescence. PMID- 8232706 TI - Focal cerebral ischemia in young adults: a collaborative case-control study. The National Research Council Study Group. AB - Because there is uncertainty about the role of atherogenic and nonatherogenic risk factors for cerebral ischemia in the young, we carried out a multicenter, hospital-based, case-control study. 333 patients (15-44 years) with focal cerebral ischemia (transient ischemic attack or stroke within 8 weeks of admission) were eligible. 25 patients were excluded, according to the protocol. 308 cases were matched by age and gender to one hospital and one population control. Independent risk was shown by logistic conditional regression for migraine with aura [odds ratio (OR) = 14.8], smoking (OR = 3.7), alcohol (OR = 2.8), serum triglycerides (OR = 1.6), arrhythmias (OR = 9.5), mitral stenosis (OR = 56), coronary heart disease (OR = 4.3) and carotid stenosis or occlusion (OR = 41). Serum HDL-cholesterol had a relative protective effect (OR = 0.8). These data confirm the role of atherosclerosis and cardiac diseases as well as migraine with aura and alcohol consumption in the pathophysiology of cerebral ischemia in the young. More thorough prevention programs may contribute to earlier detection and control of all of these risk factors, but further investigations in patients with as yet unidentified risk factors are warranted because the above-mentioned factors do not account for the total risk of ischemic stroke in the young. PMID- 8232707 TI - Incidence of stroke in young adults in the Reggio Emilia area, northern Italy. AB - A retrospective epidemiological study on the first episode of stroke in young adults aged 15-44 years was carried out in the territory of the Local Health Unit No. 9 in Reggio Emilia (46,491 km2), Italy, from 1987 to 1989. 29 patients were identified: 17 were affected with cerebral infarction and 12 with hemorrhage. All young patients were discharged with diagnostic codes 430-438 according to the International Classification of Disease, i.e. the criteria of the World Health Organisation for stroke definition. All patients had computed tomography or necropsy. The average annual incidence rate per 100,000 population aged 15-44 for all strokes was 13.6 and the 95% confidence interval (CI 95%) was 9.1-19.6. The general population of the same age on January 1, 1987, was 69,845 and 71,920 on December 31, 1989; the incidence rate of stroke was 14.0 for males (CI 95% 7.9 2.3) and 13.2 (CI 95% 7.1-22.2) for females. The average annual incidence rates were 8.0 (CI 95% 4.7-12.2) for cerebral infarction (8.4, CI 95% 3.9-16 for males, 7.6, CI 95% 3.3-14.9 for females), 5.6 (CI 95% 2.9-9.9) for cerebral hemorrhage and 2.8 (CI 95% 1.0-6.1) for both subarachnoid (SAH) and intracerebral hemorrhage (ICH). Based on angiography or necropsy findings, aneurysms or arteriovenous malformations were present in 83% of the patients with SAH and in 66% of the patients with ICH. The 1-month fatality ratio was 0 for cerebral infarction, 50% for SAH and 33% for ICH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232708 TI - Epidemiology of spinal cord injury in New Zealand. AB - Spinal cord injury (SCI) is a catastrophic and costly result of both intentional and unintentional injury. We present data from the Health Statistics Services files of New Zealand for the year 1988 on the epidemiology of SCI resulting in morbidity. New Zealand has one of the highest rates of SCI in the western world and since 1979 this has been increasing. It occurs most often to young, caucasian men and is typically the result of motor transport crashes. The ethnicity adjusted rates show high rates for Maori males. Children in New Zealand have greater than 4 times the risk of an SCI than American children. The rehabilitation and hospital costs for SCI are among the highest for all injuries. There were a higher number of high cervical injuries than reported in previous series and 92% of SCI were incomplete indicating the high rehabilitation potential of the sample. Some measures are suggested to reduce the incidence of SCI. PMID- 8232709 TI - Chronic symmetric polyneuropathy in the elderly. A field screening investigation in two regions of Italy: background and methods of assessment. The Italian General Practitioner Study Group (IGPSG). AB - There are no data on the prevalence and other epidemiologic characteristics of chronic symmetric polyneuropathy (CSP) in the community. This study was designed to assess the prevalence and risk factors of CSP in well-defined elderly (55 years and older) populations from two separate areas in Italy. Thirty-five general practitioners working in the two areas were invited to interview a random sample of patients requesting consultation. They used a pretested questionnaire which focused on selected symptoms of polyneuropathy, common risk factors for CSP, and current drug treatments. Patients giving affirmative answers to the questions on screening symptoms received a further clinical evaluation by a neurologist. Details are given of the general characteristics of the study areas and populations, the methods of case ascertainment, the validation of the screening instruments, and the results of a pilot study. PMID- 8232710 TI - [Recent aspects of the action of aldosterone]. AB - As described for other steroid hormones, aldosterone action in its target cells involves the binding to intracellular receptors, which modulate the expression of some genes. Recent progress in the field involves: the molecular structure of the mineralocorticoid receptor (MR) which has been cloned; the effects of MR on transcription; the hetero-oligomeric structure of MR; its tissue-specific distribution; the identification of 11 beta-hydroxysteroid dehydrogenase as an enzyme responsible for mineralocorticoid specificity. Finally, the effects of aldosterone at the cellular and molecular level are now better known. From the precise integrative view of its action on the kidney, the recent fields of this research extend to cellular and molecular understanding of aldosterone action. PMID- 8232711 TI - [Protein metabolism during nephrotic syndrome. Experimental and clinical influence of dietary protein intake]. AB - In adults, 12 to 14 g of albumin are synthesized daily. The same quantity is catabolized, essentially by the vascular endothelium and to a lesser degree in renal tubules. During nephrotic syndrome, contrary to what is observed in malnutrition conditions accompanied by hypoalbuminemia, hepatic synthesis is only moderately increased, whereas fractional catabolism is greater than normal. The increase in alimentary protein-intake, which has been proposed to restore the pool of albumin, raises hepatic albumin synthesis, but also its urinary losses, most likely by stimulation of the renin-angiotensin system. A reduction in protein rations lowers proteinuria and improves the hepatic abnormalities of nephrotic syndrome but its longterm nutritional consequences are poorly understood. The association of a converting enzyme inhibitor with a diet moderately restricted in protein content (1 g/kg/day) might constitute a therapeutic satisfactory solution. PMID- 8232712 TI - [Hypokalemia without arterial hypertension by licorice poisoning]. AB - We report a case of chronic intoxication with glycyrrhizinic acid, at a dosage of 1000 to 1500 mg per month over a period of 11 months, in a former alcoholic. This intoxication was revealed by profound hypokalaemia and rhabdomyolysis. However, blood pressure remained constantly normal. Analysis of the literature shows that liquorice intoxication, which blocks renal 11 beta-hydroxysteroid dehydrogenase evolves more frequently as isolated hypokalaemia than as a picture of pseudo primary hyperaldosteronism accompanied with hypertension. Hypokalaemia with urinary potassium wasting and without hypertension should therefore lead to considering liquorice intoxication, which can be confirmed by disclosing shut-off of the renin-angiotensin-aldosterone axis, and by the increase of the urinary ratio of [cortisol metabolites (5 alpha tetrahydrocortisol + 5 beta tetrahydrocortisol)]/[cortisone metabolite (5 beta tetrahydrocortisol)] together with increase of urinary free cortisol excretion. PMID- 8232713 TI - [Glycoprotein inhibitors of urinary calculi formation]. AB - Human urine is supersaturated with respect to calcium salts especially calcium oxalate which is the major mineral phase of urinary stones. Inhibitors of crystallization prevent renal calcification in urine. Small molecular weight components account for 20% of total inhibitory activity of urine. Macromolecular inhibitors (molecular weight greater than 10 kDa) are more powerful. Recently, several macromolecules have been identified in urine, in urinary stones and in the kidney: Nephrocalcin, Tamm-Horsfall protein, Uropontin, Crystal Matrix Protein, renal Lithostathine. The role of these proteins is stressed. PMID- 8232714 TI - [What is the practical attitude toward isolated microscopic hematuria?]. AB - Practical attitude towards isolated microscopic hematuria. The finding of isolated asymptomatic microhematuria usually raises questions about the need to perform further, invasive investigations. Phase contrast microscopy of the urine sediment is a sensitive, noninvasive method that provides information on the glomerular or non glomerular origin of hematuria, as well as on its grade. This analysis, however, must be performed by an experienced technician under standard conditions. The presence of dysmorphic (i.e. glomerular) erythrocytes indicates a glomerular disease if the count is higher than 10 erythrocytes per microliter but is considered physiological if the count is below this number. In these two cases, no further investigation will be undertaken if all criteria for isolated microhematuria are verified. Conversely, isomorphic erythrocytes reveal a non glomerular origin of hematuria, which may indicate a serious urological disease. In this case, further investigations (e.g. ultrasound, urine cytology) are recommended, taking into account the age of the patient. PMID- 8232715 TI - [Laparoscopic surgery: a new alternative to abdominal surgery in renal insufficiency treated by CAPD]. AB - Six laparoscopic abdominal operations were performed in five patients with chronic renal failure with continuous ambulatory peritoneal dialysis. Coelioscopic exploration and operation disclosed several advantages including peritoneal dialysis catheter salvage, abdominal exploration in peritonitis or before catheter implantation, early dialysis after surgery, hence avoiding the need for transient hemodialysis. Furthermore, laparoscopy was reported to decrease postoperative pain and ventilatory disorders. This minimal invasive surgical technique allows rapid recovery of social and professional activities. PMID- 8232716 TI - Heat-shock inhibits protein synthesis and eIF-2 activity in cultured cortical neurons. AB - Stress, such as heat-shock, hypoxia and hypoglycemia, inhibits the initiation of protein synthesis. The effects of heat-shock on protein synthesis, eucaryotic initiation factor 2 (eIF-2) activity, protein kinase C (PKC), and casein kinase II (CKII) activities were studied in primary cortical neuronal cultures. In neurons exposed to heat-shock at 44 degrees C for 20 min, protein synthesis is inhibited by more than 80%, and is accompanied by a 60% decrease in eIF-2 activity. Steady state PKC and CK II activities were not affected by heat-shock. Vanadate (200 microM), a protein phosphotyrosine phosphatase inhibitor, partially prevented the depression of eIF-2 activity during heat-shock, and increased CKII activity by 90%. In contrast, staurosporine (62nM), a protein kinase C inhibitor, did not affect eIF-2 activity. We conclude that heat-shock causes a change in the phosphorylation/dephosphorylation of regulatory proteins leading to a depressed eIF-2 activity and protein synthesis in neurons. PMID- 8232717 TI - Characterization of adenosine receptor-mediated generation of cyclic AMP in slices of rat cerebral cortex with chronic epileptic activity. AB - Cyclic AMP accumulations elicited by adenosine analogues 2-chloroadenosine (2 CADO), R-N6-phenylisopropyladenosine (R-PIA), and N6-cyclohexyladenosine (CHA) were investigated in cortical slices of chronic iron-induced epileptic rats. Cyclic AMP accumulation was elicited 9- to 18-fold by 2-CADO and it was elicited 5- to 7-fold by either R-PIA or CHA; 2-CADO was more potent than R-PIA or CHA in eliciting cyclic AMP accumulation. The adenosine analogues elicited cyclic AMP accumulation in a dose-dependent manner, and the elicitation was inhibited by the adenosine antagonist 8-phenyltheophylline. The 2-CADO-elicited accumulation of cyclic AMP was greatly increased in the cortical region on the primary epileptic side, while the R-PIA- or CHA-elicited accumulation did not change in any cortical region. The deviation detected only in the 2-CADO-elicited accumulation of cyclic AMP may be due to the difference in relative potency for adenosine receptors of the adenosine analogues. The results suggest that adenosine receptor mediated generation of cyclic AMP is altered in the primary region of iron induced epileptic cortex, in which heterogeneous alterations in different adenosine receptor subtypes may occur in the epileptic process. PMID- 8232718 TI - Urinary excretion of bioactive amines and their metabolites in psychiatric patients receiving phenelzine. AB - Phenelzine [2-phenylethylhydrazine] (PLZ), a potent inhibitor of monoamine oxidase (MAO)-A and -B, is used widely in psychiatry. We have studied the effects of PLZ administration on urinary excretion of several bioactive amines and their metabolites in psychiatric patients. Urine samples (24-hour) were collected prior to treatment and again at 2 and 4 weeks of treatment with PLZ (30-90 mg daily in divided doses). Amines and metabolites analyzed included 2-phenylethylamine (PEA), m- and p-tyramine (m- and p-TA), phenylacetic acid (PAA), m- and p hydroxyphenylacetic acid (m- and p-OH-PAA), tryptamine (T), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), normetanephrine (NME), 3-methoxy-4 hydroxyphenylglycol (MHPG), 3-methoxytyramine (3-MT), and homovanillic acid (HVA). Levels of PEA, p-TA, 5-HT, and T were elevated during treatment with PLZ, but no significant changes in urinary excretion of the acid metabolites PAA, p-OH PAA, and 5-HIAA were observed. Urinary levels of the noradrenaline metabolites NME and MHPG were increased and decreased, respectively; a similar pattern was observed with the dopamine metabolites 3-MT and HVA. There was an elevation in levels of m-TA and a decrease in its acid metabolite m-OH-PAA during the treatment with PLZ. PMID- 8232719 TI - [3H]norharman ([3H]beta-carboline) binds reversibly and with high affinity to a specific binding site in rat liver. AB - In addition to the known binding of norharman (NH) to monoamine oxidase (MAO) and benzodiazepine (BZ) binding sites (at microM concentrations), a distinct class of high-affinity NH binding sites was discovered in rat brain. Investigations of several organs of the rat led to the discovery of high affinity binding sites in the liver, which successfully could be solubilized from P2 membrane homogenate (0.25% w/v Triton X-100). Scatchard analysis revealed an apparent KD value of 26 +/- 8 nM and a maximum number of binding sites of 11 +/- 3 pmol/mg protein (n = 14). Association kinetics showed that equilibrium was nearly reached after two hours. Dissociation was totally complete only after more than 16 hours. The MAO inhibitors examined did not influence the binding characteristics. No displacement of specific binding could be found by haloperidol. PMID- 8232720 TI - Effect of hyperbaric oxygenation on the Na+, K(+)-ATPase and membrane fluidity of cerebrocortical membranes after experimental subarachnoid hemorrhage. AB - It is reported that CNS hemorrhage causes membrane dysfunction and may exacerbate this damage as a result of secondary ischemia or hypoxia. Since hyperbaric oxygenation improves oxygen metabolism, it may reduce this membrane damage. The present study was conducted to reveal whether hyperbaric oxygenation influences membrane alteration after hemorrhage. Thirty minutes after subarachnoid hemorrhage induction, rats were treated with hyperbaric oxygenation 2 ATA for 1 hour. Rats were decapitated 2 hours after subarachnoid hemorrhage induction. Na+, K(+)-ATPase activity measurement and spin-label studies were performed on crude synaptosomal membranes. Subarachnoid hemorrhage decreased Na+, K(+)-ATPase activity. Spin label studies showed that hydrophobic portions of near the membrane surface became more rigid and the mobility of the membrane protein labeled sulfhydryl groups decreased after subarachnoid hemorrhage. Hyperbaric oxygenation significantly ameliorated most of the subarachnoid hemorrhage induced alterations. We conclude that hyperbaric oxygenation may be a beneficial treatment for acute subarachnoid hemorrhage. PMID- 8232721 TI - Henry McIlwain, 1912-1992. PMID- 8232722 TI - Trace amines and Tourette's syndrome. AB - There has been considerable interest in recent years in possible neurochemical abnormalities in Tourette's Syndrome (TS). In studies combining neuropsychological and neurochemical measurements, we have investigated the possible roles of trace amines in this disorder. Urinary levels of free beta phenylethylamine (PEA) and plasma levels of its precursor amino acid phenylalanine were decreased in TS patients when compared to values in normal children. These urinary PEA levels in TS patients were inversely related to several scores from the Tourette's Syndrome Global Scale (TSGS). Further investigation of the group of subjects with low urinary PEA indicated that they also had low levels of MHPG, normetanephrine, 5-HT and m- and p-tyramine. Patients with low PEA were also compared on an extensive battery of neuropsychological measures and observed to perform significantly worse than TS patients with normal urinary PEA levels. Biochemical measurements also suggest a possible abnormality in tryptamine turnover in TS since urinary levels of indole 3-acetic acid (IAA; the acid metabolite of tryptamine) are significantly lower in TS patients than in normal controls. PMID- 8232723 TI - Effects of cell signaling on the development of GABA receptors in chick retina neurons. AB - R-cognin, a cell recognition molecule, and insulin are known to play significant roles in GABAergic differentiation in the developing chick retina. In the present study, the effects of insulin and R-cognin on post-synaptic (GABAceptive) differentiation were investigated. In ovo binding of [3H]GABA and [3H]flunitrazepam ([3H]Flu) to the GABA and benzodiazepine (BZD) receptors, respectively, remained at low levels during early embryogenesis but increased sharply from mid-embryogenesis through hatching, increases which also occur in cultured neurons from early-embryonic (E7) and mid-embryonic (E11) chick retina. E7 neurons respond to insulin treatment (100 ng/ml) with increased [3H]Flu binding but no change in [3H]GABA binding. Cognin antibody (10 micrograms/ml) treatment of E7 neurons caused no significant inhibition of the developmental increases in binding of either radioligand. Insulin in E11 cultures led to greater developmental increases in binding sites for both radioligands, but exposure to cognin antibody was without significant effect. These data, along with previous studies, indicate that GABAergic differentiation in developing chick retina is regulated, in part, by insulin and cognin-mediated cell signaling. Insulin also regulates post-synaptic (GABAceptive) differentiation whereas cognin-mediated interactions are relatively insignificant. PMID- 8232724 TI - Dopamine as trace amine in the dorsal root ganglia. AB - It has been shown that in the chick dorsal root ganglion (DRG) about 8% of neurons, belonging to both the A and B classes of sensory neurons exhibit a clear dopamine immunoreactivity. In the present study are reported the results of measurements, by mean of HPLC-electrochemical detection (HPLC-ED), of DA and of the DA metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the rat DRG and their central nerves. Very low levels of DA, about 10 folds lower than the levels found in the dorsal horn of the spinal cord, were found in the DRG. However the levels of DOPAC and HVA were approximately equivalent to the levels found in the cord. The immunocytochemical study performed in parallel has shown that some dopaminergic-immunoreactive fibers in the DRG are located around the blood vessels. Few dopamine-immunoreactive sensory neurons were identified in the DRG and immunoreactive fibers, not linked to blood vessels, were identified in the dorsal root nerves. The present work indicates that there is a dopaminergic innervation of the blood vessels in the rat DRG but that dopamine may also be, as in the chick, a transmitter of primary afferent fibers. PMID- 8232725 TI - High ammonia diet: its effect on the glial fibrillary acidic protein (GFAP). AB - The effect of a recent hyperammonemic model, consisting of a high ammonia diet for 3, 7, 15, 45, and 90 days, on glial fibrillary acidic protein (GFAP) in the rat spinal cord and on blood ammonia levels has been studied. The high ammonia diet was prepared by mixing a standard diet with ammonium acetate (20% wt/wt); in addition, 5 mM of ammonium acetate was added to the water supply. GFAP contents were determined by means of immunoblotting analysis. The results demonstrated that this high ammonia diet model neither induces significant changes in GFAP immunoreactivity, nor modifies total protein concentration, and only induces significant blood hyperammonemic levels in the first days of treatment. An adaptive response to the diet is suggested and discussed to explain these results. A relation between ammonia and GFAP expression is suggested because transient hyperammonemia induces transient, although no significant, changes on GFAP expression. PMID- 8232726 TI - GABA-gated chloride ion influx in brains of tremor rats. AB - We measured the GABA-gated chloride ion influx and GABA concentrations in the cerebral cortex and the hippocampus of young (5 weeks old) and older (15 weeks old) tremor rats. GABA-gated chloride ion influx in these tremor rats was significantly greater than in the controls of both the 5 week- and 15 week-old groups. GABA concentrations in the cerebral cortex and hippocampus of the tremor rats increased compared with controls of 5 weeks and decreased compared with controls of 15 weeks. These findings suggest that the GABAergic presynaptic neurons in the cortex and hippocampus of the tremor rat are disturbed with aging. This change may be related to the appearance of absence-like seizures in the rats. The increased GABA-gated chloride ion influx in tremor rats may be a compensatory mechanism against the genetically-determined seizure susceptibility of these rats. Furthermore, the increased GABA levels and GABA-gated chloride ion influx found in 5 week-old tremor rats may be related to the tremor movements. PMID- 8232727 TI - Interaction of tacrine and velnacrine with neocortical synaptosomal membranes: relevance to Alzheimer's disease. AB - The acridine-based, potential Alzheimer's disease therapeutic agents, tacrine and velnacrine, were incubated with rat or gerbil neocortical synaptosomal membranes. Electron paramagnetic resonance employing a protein-specific spin label was used to monitor this interaction. Analogous to their effects in erythrocyte membranes [Butterfield and Rangachari (1992) Biochem. Biophys. Res. Commun. 185: 596-603], in the present studies both agents decreased segmental motion of spin labeled synaptosomal membrane proteins, consistent with increased cytoskeletal protein protein interactions (0.001 < P < 0.005), and tacrine was more potent than velnacrine. These results are discussed with possible relevance to molecular actions of the agents and molecular alterations in Alzheimer's disease. PMID- 8232728 TI - Determination of regional distributions of phenylethylamine and meta- and para tyramine in rat brain regions and presence in human and dog plasma by an ultra sensitive negative chemical ion gas chromatography-mass spectrometric (NCI-GC-MS) method. AB - Using a new ultrasensitive method the trace biogenic amines, phenylethylamine, meta-tyramine and para-tyramine have been quantitated in brain regions obtained from a single rat. Phenylethylamine concentrations in ng/g wet tissue (mean +/- std. error) were as follows: caudate 2.71 +/- 0.73, hypothalamus 0.45 +/- 0.15, cerebellum 0.09 +/- 0.02, olfactory bulb 0.35 +/- 0.11, stem 0.13 +/- 0.03, hippocampus 0.20 +/- 0.11, cortex 0.69 +/- 0.13 and the rest (remainder of the brain) 2.81 +/- 0.41. Mean whole brain was 1.23 +/- 0.19 ng/g, in agreement with previous measurements. meta-Tyramine concentrations (ng/g) were: caudate 2.69 +/- 0.19, hypothalamus 0.32 +/- 0.16, cerebellum 0.07 +/- 0.04, olfactory bulb 0.09 +/- 0.04, stem 0.04 +/- 0.01, hippocampus 0.07 +/- 0.02, cortex 0.18 +/- 0.15 and the rest 0.15 +/- 0.06, with a mean whole brain value of 0.26 +/- 0.05 ng/g and para-tyramine concentrations were: caudate 8.99 +/- 1.60, hypothalamus 0.93 +/- 0.13, cerebellum 0.78 +/- 0.27, olfactory bulb 0.70 +/- 0.13, stem 0.90 +/- 0.36, hippocampus 0.40 +/- 0.06, cortex 1.78 +/- 0.28 and the rest 2.38 +/- 0.12 and mean whole brain was 1.90 +/- 0.25 ng/g. In human plasma the concentrations of the three amines were found to be 31.3 +/- 3.4 pg/ml, 5.3 +/- 1.6 pg/ml and 66.0 +/- 9.9 pg/ml respectively and in dog blood 95.3 +/- 4.6 pg/ml, 24.0 +/- 7.6 pg/ml and 486 +/- 43 pg/ml respectively. When monoamine oxidase inhibitors were added to the blood immediately after collection there were no significant increases in the amine levels indicating that MAO-B is not present in plasma in significant quantities. PMID- 8232729 TI - [Th usefulness of the Mini-Mental State Examination in quantitative estimation of the severity of cognitive impairment in patients with multifocal vasogenic cerebral lesions]. AB - The material studied comprised 65 inpatients admitted to the Department of Cerebrovascular Diseases from January 1988 to September 1990. The patients had a history of multiple episodes of brain ischaemia confirmed by CT scan, and had clinical evidence of focal symptoms and signs clinically judged to the related to disturbances in cerebral circulation. The aim of this study was a comparison of usefulness of the Mini-Mental State Examination and other clinical screening scales in estimation of the cognitive impairment among these patients. We performed an analysis of the results on MMSE in the studied persons in comparison with the results in Dementia Scale (SD). The clinical stage of dementia was assessed by I part of the Brief Cognitive Rating Scale (BCRS). After the first selection material was divided into 2 groups: 1. group I (34 patients) included patients scoring 23 or less in the MMSE, 2. the remaining patients formed group II (31 persons). They scored 24 to 29 in the MMSE and complained about memory loss. The results of the SD and BCRS demonstrated the following: persons with high scores in the MMSE had intellectual impairment in other screening scales. The observations are consistent with the opinion of other authors. They do not doubt the usefulness the MMSE but regard it as an indicatory test for grading of cognitive impairment. They underlined the fact, that the majority of the patients even with higher scores in Mini-Mental State Examination may have moderate or even severe dementia. PMID- 8232730 TI - [Follow-up examinations of patients with a history of reversible ischemic stroke]. AB - Follow-up examinations are reported of 218 patients with a history of reversible ischaemic stroke. The examinations were carried out 4 years after the onset of the disease which led to first hospitalization (1984-1987). It was possible to determine the frequency of repeated ischaemic episodes which developed in 27.5% of cases, including secondary transient ischaemic attacks--13.3%, reversible ischaemic strokes--4.1%, ischaemic strokes with permanent consequences--10.1%. In most cases (75%) they occurred in the first year after the first episode. PMID- 8232731 TI - [High dose methylprednisolone therapy in patients with multiple sclerosis]. AB - A trial of high-dose pulsed intravenous methylprednisolone was carried out in 20 patients with multiple sclerosis (MS). Multimodal evoked potentials examination (visual, brainstem auditory and somatosensory evoked potentials) were evaluated before and after treatment with methylprednisolone (a single daily dose of 1000 mg was administrated for five consecutive days). Clinical improvement did not correlate well with evoked potential changes. PMID- 8232732 TI - [Neuroborreliosis: diagnostic difficulties]. AB - Borreliosis is a condition caused by spirochaete-Borrelia burgdorferi, characterised by multiorgan changes and prolonged course with three consecutive stages. The number of diagnosed cases in Poland tends to increase. This is due to the availability of diagnosis tests. Owing to the analysis of blood and CSF by the fluorimetric method 3M IgG/IgM Fastlyme, neuroborreliosis was confirmed in 5 patients (hospitalised in Neurological Clinic of Medical Academy in Bialystok). Three of the cases were characterised by a bilateral palsy of facial nerve and abnormal CSF, as evidenced by pleocytosis (100% lymphocytes) and elevated protein. Patient 4 diagnosed by dermatologists as having Acrodermatitis atrophicans, should be classified, however, as the third stage of the disease, while according to neurological changes--namely a lesion of shoulder plexus--as the second stage. Evaluation of patient 5 presented most difficulties. Considering long-term disease and the time from the onset to the admission to the neurological clinic, she was likely to develop the third stage of the disease, and therefore, our treatment failed to be so effective as in aforementioned cases. PMID- 8232733 TI - [Evaluation of changes in the epidemiological situation of SSPE in Poland in the years 1977-1991 (in the course of 15 years after the introduction of mandatory antimeasles vaccination)]. AB - The authors present the results of an epidemiological study of SSPE in Poland conducted since 15 years. Since the years covered by the study (1977-1991) included the first period after the introduction of obligatory vaccinations against measles in our country, the most important aim of this analysis was to find out the possible influence of the effect of these vaccinations on SSPE incidence. A comparison of the data obtained in successive stages of the study shows that a slow gradual decrease of the annual incidence of SSPE has been taking place in our country (from at least 1.6 per million before 1983 to 0.69 in 1991). The effect of the vaccinations is visible also in a significant increase of the mean age of SSPE onset which is a result of ever more widespread vaccination of the youngest children. These observations indicate unequivocally that the aetiological factor in SSPE is a wild strain of measles virus and not that used for vaccinations. PMID- 8232734 TI - [Changes of sensory threshold of nerve fibres in polyneuropathies]. AB - Subjective and objective sensory threshold (Ts, T(ap)) was measured in the sensory fibres of the median and the sural nerves of patients with diabetic neuropathy, Guillain-Barre-Strohl syndrome, hereditary motor and sensory neuropathy (HMSN I and II) as well as in the carpal tunnel syndrome (CNS). A very marked increase of the Ts was found in HMSN I in both nerves investigated. In HMSN II significant increase of Ts were present only in the sural nerve. In CTS significant increase of Ts was found. Measurements of Ts may be useful in the evaluation of the intensity of sensory nerves lesion. PMID- 8232735 TI - [Comparison of two methods of postoperative radiotherapy in anaplastic cerebral oligodendroglioma]. AB - Preliminary results of prospective clinical study--49 adult patients with histologically proven anaplastic oligodendroglioma received postoperative radiotherapy. Patients were randomized to two treatment arms: Conventionally Fractionated Radiotherapy (CFR) and Large Fractionated Radiotherapy (LFR). In CFR group the whole brain was irradiated to the total dose of 50 Gy in 25 fractions over 5 weeks, then a 10 Gy "boost" in 5 fractions was delivered to the site of primary lesion in 5 days. In LFR group there were 3 courses of irradiation separated with one month interval. In each of the two first series the patients received 20 Gy in 5 fractions in five days to the whole brain, and in the third course, 10 Gy "boost" was delivered like in CFR regimen. The tolerance to treatment was good in both groups. Actuarial survival rates 2 and 4 years were 20% and 4% respectively for CFR group, and they were 27% and 0% respectively for LFR. PMID- 8232736 TI - [Results of surgical treatment of retrobulbar tumors through osteoplastic supraorbital osteotomy]. AB - Osteoplastic supraorbital orbitotomy in operations of retrobulbar tumours minimizes the necessity of retraction of frontal lobe by change of the direction of the surgical approach. The method allows also reconstruction of the orbital roof preventing postoperative pulsating exophthalmos. The results were supported by comparison of the late control CT scans performed in patients operated by the classical transcranial approach with the examinations performed in patients operated on by the described method. PMID- 8232737 TI - [Arteriovenous malformation as the most frequent form of cerebral vascular dysplasias]. AB - The purpose of the study was a survey of autopsy and biopsy material from 27 cases of cerebral vascular malformations for establishing the occurrence of various forms of these lesions in relation to that reported from other centres. It was tried also to check in the light of the morphological analysis of our cases the theory of vascular malformation development put forward by Kaplan et al (1961). The material comprised 4 cases of telangiectasia, 11 arteriovenous malformations, 7 cavernous haemangiomas and 5 mixed cavernomas with arteriovenous malformation. Arteriovenous malformations prevailed. The clinical manifestations set in at the age between 15 and 46 years. The analysis rather confirm the concept of Kaplan et al. that dysplasia is derived from disturbances in the development of capillaries in early embryogenesis. All other comparisons and observations were consistent with those reported in literature outside Poland. PMID- 8232738 TI - [Neurotransplantations: experimental studies]. AB - Before the intracerebral transplantations of adrenal medulla and fetal substantia nigra in parkinsonian patients were introduced to restore the dopamine deficit, many intensive experimental studies were performed during the last decades. The history of these experiments was presented including the animal models of Parkinson's disease, different implantation techniques and the effects of neurotransplantations of adrenal medulla and fetal substantia nigra obtained in experimental animals. Different mechanisms underlying the transplant-induced functional effects as well as the methods of its stimulation to extend the graft survival and function were described. The ability of intracerebral transplantations to ameliorate the symptoms in animal models of disorders other than Parkinson's disease is also discussed. PMID- 8232739 TI - [Central nervous system tuberculosis]. AB - On the basis of a literature review the author discusses the aetiology, pathogenesis, diagnosis, clinical course and treatment of central nervous system tuberculosis. Attention is called to the necessity of further improvement of diagnostic methods for making possible early diagnosis and modern treatment of this disease. PMID- 8232740 TI - [Intervertebral disc surgery: intraoperative complications]. AB - Several hundreds of thousands of operations on the lumbosacral spine are carried out annually for decompression of nerve roots and disc removal. It is the opinion that serious intraoperative complications developed in 2.5 to 13.7% of cases. In the few literature reports cases have been described of dural sac rupture, injury to large retroperitoneal vessels, intestinal perforation and complete severing of ureters. PMID- 8232741 TI - [A case of coexistence of hypophyseal adenoma and epileptic seizures]. AB - A case of hypophyseal adenoma with epileptic seizures in a man aged 61 years is reported. The generalized seizures developed 21 years earlier, and partial complex seizures 2 years before the diagnosis of adenoma. The patient had two sons, one died of suprasellar tumour, the other one has generalized and partial complex seizures. PMID- 8232742 TI - [A case of dystrophic myotonia with familial analysis]. AB - The description of the 37-years old patient is a classic example of fully symptomatic myotonic dystrophy. The diagnosis was established on: 1. the medical history of the patient, 2. the analysis of pedigree which revealed the treatment of 8 cases of the disease, 3. the examination which demonstrated central signs of myotonic dystrophy, 4. laboratory investigations especially the histopathological examination of the tibial anterior muscle and electromyography. The analysis of the pedigree confirmed the genetical background of the disease and its autosomal dominant inheritance. The 4-year follow-up confirmed the progressive course of the disease. PMID- 8232743 TI - [Multiple venous air embolism during cerebellar tumor surgery. Case report]. AB - A case of successfully treated multiple venous air embolism occurred in a man aged 41 operated on for cerebellar tumour. Postoperatively exceptionally serious complication developed in the form of neurogenic pulmonary oedema. Attention is called to a number of aetiological factors in this case. The anaesthesiological management in such cases should include careful monitoring of end-tidal CO2 concentration and insertion of catheter into right atrium. PMID- 8232744 TI - [Sacral meningeal cyst as a cause of chronic pain related to perineal, sacral and pelvic regions]. AB - The authors present a case of sacral meningeal cyst (s.m.c.) being the cause of chronic pain related to perineal, sacral and pelvic regions. The aim of the report is to emphasize the role played by s.m.c. in the aetiology of pain arising in these regions. As it can result from a wide variety of pathologies, like gynaecological, urological and anorectal the differential diagnosis is very difficult. Until recent years s.m.c. was rarely encountered in clinical practice and identified as a cause of chronic pain in perineal, sacral or pelvic areas. Now in the era of MRI and with increasing access to sophisticated imaging diagnostic methods s.m.c. will probably be recognized in more cases of perineal, sacral or pelvic pain. PMID- 8232745 TI - [Brief considerations of the causes of ischemic stroke in relation to patients' age]. PMID- 8232746 TI - [Discussion of a possibility of oncologic sequences of the Chernobyl disaster]. PMID- 8232747 TI - [Discussion of an undiagnosed tumor of foramen occipital magnum]. PMID- 8232748 TI - [Konigsmark-Hollander-Berlin epilepsy?]. PMID- 8232749 TI - Parvalbumin and calbindin D-28k in the human motor system and in motor neuron disease. AB - Calbindin D-28k and parvalbumin are neuronal calcium binding proteins of interest in relation to neurodegenerative diseases. Expression of calbindin and parvalbumin may be one of the determinants of selective vulnerability in these disorders. The distribution of these proteins was surveyed in the normal human motor system and in motor neuron disease (MND) using immunocytochemistry in formalin fixed post-mortem tissues. CNS tissues from 14 MND patients (mean age 61.2 years, mean post-mortem delay 24.6 h) and seven controls (mean age 62.6 years, mean post-mortem delay 25.3 h) were studied. Preliminary studies on the effects of fixation were performed. In normal cases upper and lower motor neurons showed absent expression of both proteins. Several neuronal groups characteristically spared in MND showed varying patterns of immunoreactivity: oculomotor neurons showed parvalbumin staining of the perikaryon; the thoracic preganglionic sympathetic neurons showed calbindin staining in perikarya. Onuf's nucleus showed calbindin staining in the neuropil only. In motor neuron disease a loss of ventral horn interneurons and calbindin immunoreactive processes was observed with no other disease related changes in the spinal cord, brain-stem, or motor cortex. These findings are consistent with the hypothesis that the distribution of these proteins is one determinant of selective vulnerability to the neurodegenerative processes in MND acting via disturbance of neuronal calcium homeostasis. PMID- 8232750 TI - Granule cells and the development of synapses with mossy fibres in the cerebellum of the jaundiced Gunn rat. AB - The development of granule cells and the formation of synapses with mossy fibres were investigated in cerebella from jaundiced (j/j) Gunn rats with hereditary hyperbilirubinemia. The external granular layer was thinner than that in cerebella from heterozygous (+/j) Gunn rats without hyperbilirubinemia on days 12 and 15 after birth, and it had disappeared in places by day 18. Migrating granule cells decreased in number from days 12 to 18. In the internal granular layer, the paucity of granule cells became apparent on day 12 and a reduction in the number of cells was prominent from day 18 onward. More degenerating cells were found in j/j cerebella than in +/j cerebella on days 12 and 15. Mossy fibre-granule cell synapses (glomeruli) were immature in appearance in j/j and +/j cerebella from days 7 to 12; small, roundish mossy fibre terminals were in contact with a few dendrites of granule cells. After day 18, the glomeruli were mature in terms of shape in +/j cerebella. By contrast, the mossy terminals remained small and roundish, and the granule cell dendrites that surrounded the mossy terminals did not increase in number nor develop digitiform branches in j/j cerebella. The mossy terminals were partially surrounded by glial processes and were occasionally surrounded by large dendrites of Golgi cells, which made multiple synaptic contacts. Some degenerating mossy terminals were present from days 18 to 30. These anomalous glomeruli may be caused by insufficient synaptic contacts with granule cells. PMID- 8232751 TI - Cytoplasmic expression of the leu-4 (CD3) antigen in developing Purkinje cells in the rat cerebellum. AB - Although commonly known to represent a T cell receptor (CD3) associated polypeptide, the leu-4 (CD3) antigen occurs in cerebellar Purkinje cells (PCs) of many species. The monoclonal pan T lymphocyte marker anti-leu-4 (CD3) recognizes both the lymphocytic and the Purkinje cell type of this antigen [22]. To obtain more information about the merit of anti-leu-4 (CD3) as an investigational tool, we evaluated the expression of leu-4 (CD3) in PCs of the developing rat cerebellum (in situ) by light microscopy. Positive anti-leu-4 (CD3) immunoreaction of PCs did not occur prior to post-natal day (D) 4. The analysis of immunostaining during cell differentiation revealed three major phases of post natal PC maturation including antigenic development of cell somata (phase 1: until D6), dendrites (phase 2: D7-D11), and axons (phase 3: D12-D14). A massive post-weaning expansion of the dendritic arborization led then to the mature PC architecture. Additionally, the leu-4 (CD3) antigen was observed in ectopic PC dendrites (D10) and in ectopic (mature) PCs. Throughout post-natal development as well as in mature PCs, the leu-4 (CD3) antigen was found to be cytoplasmic. Due to its labile nature, neither an ultrastructural localization nor molecular characterization could be achieved. For the same reason, its application is basically restricted to cryo-fixed cerebellar tissue. However, at the level of light microscopy, the monoclonal human T cell marker anti-leu-4 (CD3) proved to be a useful tool for specific and sensitive labelling of differentiated cerebellar PCs in the rat. PMID- 8232752 TI - Ependymal cells of the choroid plexus express tumour necrosis factor-alpha. AB - Tumour necrosis factor-alpha (TNF alpha) is a major proinflammatory cytokine which appears in the cerebrospinal fluid very early after endotoxin challenge, and is likely to be produced locally. Following in vivo and in vitro challenge with endotoxin, we have demonstrated immunocytochemically and by in situ hybridization that pig and guinea-pig choroid plexus ependymal cells can produce TNF alpha. Immuno-electron microscopy shows that this protein is localized within ependymal cells to the cytoplasm and microvilli. We suggest that this TNF alpha may be important in the initiation of the inflammatory response in bacterial meningitis. PMID- 8232753 TI - Astroglial changes in the cerebral cortex of AIDS brains: a morphometric and immunohistochemical investigation. AB - Astroglial changes in the cerebral cortex of AIDS brains were analysed by means of morphometry. Astrocytes with and without immunoreactivity for glial acidic protein (GFAP) were counted and their size was measured. In the two investigated cortical areas (frontal and parietal), a similar reaction pattern of astroglia was observed. The total number of astrocytes (i.e. GFAP-positive and GFAP negative astrocytes) did not differ between control and AIDS brains. However, the number of GFAP-positive astrocytes was significantly increased in AIDS brains, while the number of GFAP-negative cells was significantly reduced. Nuclear size of GFAP-negative and GFAP-positive astrocytes was significantly increased. The reaction pattern of cortical astrocytes in AIDS seems to be characterized by GFAP production in protoplasmic astroglia as well as by hypertrophy of all astrocytes. PMID- 8232754 TI - Different patterns of neuropathological disease in rhesus monkeys infected by simian immunodeficiency virus, and their relation to the humoral immune response. AB - The brains of 21 rhesus monkeys inoculated with SIVMAC251 were examined after intervals ranging from 3 to 27 months and compared with five uninoculated controls. Eighteen animals became infected and individually exhibited several distinct patterns of disease. Nine (50%) had largely intramural leptomeningeal venous infiltrates (LMVI) without multinucleate giant cells (MGC) or foamy macrophages. Three (17%) had only MGC lesions, involving the cerebral parenchyma. One had both patterns and five (33%) neither. The controls had sparse and tiny LMVI only, similar to three inoculated animals that did not become infected. Immunohistochemistry showed the predominance of T and B lymphocytes in LMVI and choroid plexus mononuclear lesions but a predominance of macrophages over lymphocytes in the MGC lesions. Specific disease patterns differed in their association with the humoral immune response. Animals with LMVI were all hypergammaglobulinaemic when killed compared to pre-inoculation levels, and the size of the change in serum immunoglobulin concentration was positively correlated with a quantitative index of LMVI density. Furthermore, their post mortem lymph node histology was hyperplastic. In contrast, animals found at autopsy to have MGC brain lesions were hypogammaglobulinaemic compared to preinoculation. The results are consistent with two phases in SIV-associated disease: one characterized by LMVI and hypergammaglobulinaemia and another featuring MGC and hypogammaglobulinaemia. PMID- 8232755 TI - Absence of neuron-associated microtubule proteins in the rat C-6 glioma cell line. A comparative immunoblot and immunohistochemical study. AB - Three neuron-associated microtubule proteins, Class III beta-tubulin isotype, MAP 2, and tau, were evaluated in a comparative immunoblot and immunohistochemical study of the rat C-6 glioma cell line maintained for up to 31 days in vitro. Western blots on whole SDS extracts of cells grown: (i) as monolayers on plastic dishes (for 13 and 16 days); (ii) as monolayers on poly-D-lysine coated glass coverslips (for 3, 7, and 11 days); and (iii) as explants on Gelfoam matrices (for 10, 30, and 31 days) were probed with monoclonal antibodies (MoAb) specific for the above-mentioned microtubule proteins. For these and all other markers employed, immunoperoxidase histochemistry was performed only on the matrix cultures. The immunoblot experiments demonstrated that the Class III beta-tubulin isotype, MAP2, and tau were not expressed by the C-6 cell line in any of the culture conditions, nor were they found by immunohistochemistry. In contrast, explants from all culture conditions were positive for glial fibrillary acidic (GFA) protein and for a universal anti-beta-tubulin isotype MoAb by immunoblotting, as well as by immunohistochemistry in Gelfoam matrix cultures maintained in an organ culture system. Both sets of experiments indicate that these markers are not altered under three different conditions of growth over a one-month period in vitro. The expression of GFA protein and the absence of detectable levels of Class III beta-tubulin, MAP2, and tau are in keeping with the astrocytic phenotype of the C-6 cell line. PMID- 8232756 TI - Fusion of myogenic cells to the newly sealed region of damaged myofibres in skeletal muscle regeneration. AB - In regenerating skeletal muscle, sarcoplasmic extensions containing variable numbers of nuclei, widely referred to as 'buds' or 'stumps', are formed at the ends of damaged myofibres. In this paper we investigated whether the nuclei seen in the buds results from fusion of myogenic cells or from migration of myonuclei to the sealed ends of damaged myofibres in murine muscle regenerating after crush injury. The fusion of mononuclear and multinucleate myogenic cells to the buds was demonstrated by transmission electron microscopy. In order to elucidate the frequency and kinetics of cytoplasmic continuity between myotubes and sealed myofibres, we labelled the damaged myofibres with carbocyanine dye DiI (which inserts into the lipid bilayer and travels down continuous membranes) and the samples were then examined by confocal scanning microscopy. This technique showed that there was little fusion between myotubes and myofibres during the first 6 days after crush injury, but significant fusion had occurred by the tenth day especially at the newly sealed region of the damaged myofibre. A scheme for the repair of damaged skeletal muscle is presented. PMID- 8232757 TI - Distinction between artefactually shrunken and truly degenerated 'dark' neurons by in situ fixation with microwave irradiation. AB - Dark, shrunken neurons frequently occur as artefacts in immersion fixed tissue. Perfusion fixation will prevent artefacts of this type. However, morphologically identical neurons have been described as truly degenerated cells in perfusion fixed brains in various pathological conditions. Since adequate perfusion is difficult to obtain in some pathological conditions, the question still remains whether the dark neurons found in some of these situations are true in vivo phenomena or artefacts caused by inadequate fixation. In the present study rat brains with cryogenic lesions were fixed in situ by microwave irradiation. With this method no artefactually changed dark neurons were observed in the normal parts of the brains. In the cryogenic lesions, however, a narrow rim of dark, shrunken neurons occurred adjacent to the normal cortex. This zone was identical to that observed in perfusion fixed tissue. Since inadequate fixation due to uneven perfusion of the damaged tissue is prevented with this method, we suggest that the neuronal changes represent true in vivo phenomena. Fixation with microwave irradiation can thus be used to differentiate between artefactually changed and truly degenerated dark neurons in various pathological conditions. PMID- 8232758 TI - Pituitary corticotroph function during the stress hyporesponsive period in neonatal rats. AB - Rats undergo a period in early postnatal development during which they exhibit a reduced response to stress (stress-hyporesponsive period, SHRP). SHRP is marked by a reduced capacity to secrete adrenocorticotropin (ACTH) and corticosterone in response to stressful stimuli. To characterize pituitary responsiveness during this period, we examined the release of ACTH from cultured pituitary cells derived from rats at different ages. Corticotrophs were studied from animals before (day 1 postnatally), during (days 6 and 11) and after (days 16, 20 and adult) SHRP. Direct stimulation of dispersed pituitary cells, with increasing concentrations of synthetic corticotropin-releasing factor (CRF; 1 x 10(-12) to 1 x 10(-8) M), produced significant ACTH release at all ages tested. The greatest maximal release was observed for cells isolated from animals at 6 and 11 days of age (in the midst of SHRP). CRF-stimulated ACTH release from cells obtained just prior to (day 1) or immediately following (days 16 and 20) the hyporesponsive period was dramatically reduced (relative to cells from adult animals). SHRP is, therefore, not due to diminished intrinsic corticotroph responsiveness. In fact, pituitary cells derived from animals in SHRP exhibited enhanced ACTH secretion relative to cells from animals outside SHRP. This enhanced secretion was manifested as both an increased basal (unstimulated) release of ACTH as well as an increased maximal release of ACTH at saturating concentrations of CRF. PMID- 8232759 TI - Time course of altered thyroid states on 5-HT1A receptors and 5-HT uptake sites in rat brain: an autoradiographic analysis. AB - Although a link between the hypothalamic-pituitary-thyroid (HPT) axis and affective disorder has been established, the mechanism underlying this relationship remains unclear. Since the serotonin (5-HT) system appears to be involved in the pathophysiology of mood disorders, the time course of the effects of thyroidectomy (TXT) with or without thyroxine (T4) replacement on 5-HT1A receptors and 5-HT uptake sites was examined. TXT caused a significant increase in 3H-8-hydroxy-2-(di-n-propylamino)-tetralin (3H-DPAT) binding to 5-HT1A receptors in the cortex and hippocampus at 7 days and this increase was also evident at 35 days following TXT. By contrast, TXT did not have a significant effect on 3H-DPAT binding in the hypothalamus or in the dorsal raphe nucleus. TXT did not affect the binding of 3H-cyanoimipramine (3H-CN-IMI) to 5-HT uptake sites in any of the brain regions analyzed, or at any of the time points studied. Administration of high-dose T4 for 28 days caused the binding of 3H-DPAT to recover to sham levels in the cortex, to increase in the hippocampus and hypothalamus, and had no effect in the dorsal raphe nucleus. Replacement with high-dose T4 had no effect on 3H-CN-IMI binding to 5-HT uptake sites when compared to sham-operated animals at all time points examined. These results suggest that a neuromodulatory link may exist between the HPT axis and 5-HT1A receptors in the limbic regions of the rat brain. Depending on the brain region examined, a differential response to circulating levels of thyroid hormone was observed. PMID- 8232760 TI - Loperamide inhibits corticotrophic cell function by a naloxone-insensitive mechanism in the rat in vitro. AB - The effect of the antidiarrheal drug loperamide, a mu-opiate agonist, on ACTH secretion and biosynthesis, cAMP generation and phosphoinositide turnover was studied in rat anterior pituitary cell cultures. The cAMP-dependent protein kinase A pathway was stimulated with both corticotropin-releasing hormone (CRH; 2 5 nM) and the membrane-permeable Bu(2)cAMP (0.5-2.5 mM). The protein kinase C pathway was stimulated with 1 microM arginine vasopressin (AVP) and 1-10 nM phorbol 12-myristate 13-acetate (PMA). After 3.5 h, loperamide (10 microM) had no effect on basal ACTH levels but significantly suppressed CRH-induced ACTH release, in a dose-dependent manner, to 60 +/- 4% of control (100%) (p < 0.0001). After 24 h, basal proopiomelanocortin mRNA was significantly decreased to 50% of control by loperamide (p < 0.05). The suppressive effect of loperamide on CRH induced ACTH secretion was not reversible by naloxone (0.1-1,000 microM). Morphine (0.01-10 microM) had no effect on basal and CRH-induced ACTH secretion. Loperamide did not influence basal and CRH-induced adenylate cyclase activity in anterior pituitary cell membrane preparations, but it significantly blunted Bu(2)cAMP-induced ACTH secretion in cell culture from 100 +/- 4 to 77 +/- 4% (p < 0.05). In Ca(2+)-depleted medium (Ca2+ < 0.1 mM), loperamide had no suppressive effect on CRH-induced ACTH secretion. AVP-induced ACTH secretion was significantly suppressed by loperamide from 100 +/- 5 to 74 +/- 3% (p < 0.0001), while basal and AVP-induced inositol 1-phosphate generation and PMA-induced ACTH secretion were not affected by loperamide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232761 TI - gamma-Aminobutyric acid-induced elevation of intracellular calcium concentration in pituitary cells of neonatal rats. AB - Gamma-aminobutyric acid (GABA) increased intracellular calcium concentration ([Ca2+]i) of newborn rat pituitary cells in suspension measured by the FURA-2 method. The effect of GABA was dose dependent in the range of 0.1-10 microM. This effect diminished with postnatal age as measured at days 2, 14 and 21, and in adult animals. The GABA stimulation was mimicked by muscimol; in contrast, baclofen (up to 100 microM) was ineffective. Picrotoxin, a GABAA antagonist interacting with GABA-activated chloride ionophores, caused a dose-dependent inhibition of the [Ca2+]i elevating effect of 100 microM GABA or muscimol. These observations indicate the involvement of GABAA type receptors. The GABA or muscimol effect on [Ca2+]i was antagonized by nifedipine (10 microM) or verapamil (50 microM), and completely abolished in the presence of 4 mM EGTA (low-calcium medium). The findings indicate the presence of depolarizing GABAA receptors on neonatal rat pituitary cells. It seems very likely that the mechanism by which GABA receptor occupation results in elevated [Ca2+]i is a membrane depolarization by increased Cl- conductance followed by calcium influx through L-type voltage dependent calcium channels. PMID- 8232762 TI - Impaired histamine- and stress-induced secretion of ACTH and beta-endorphin in vasopressin-deficient Brattleboro rats. AB - Arginine vasopressin (AVP), corticotropin-releasing hormone (CRH) and catecholamines seem to be involved in the histamine- (HA) and/or stress-induced release of the pro-opiomelanocortin-derived peptides adrenocorticotropic hormone (ACTH) and beta-endorphin (beta-END). The AVP component of the regulatory mechanism can be specifically studied in Brattleboro rats which lack AVP. These animals may therefore serve as a useful biological model for investigating the importance of AVP in the ACTH and beta-END response to HA and stress. On this background, we studied the ACTH and beta-END response to HA or restraint stress in conscious, male dizygotic AVP-deficient Brattleboro rats (DI) and compared the hypothalamic content of CRH and catecholamines in these rats with that of nondiabetic isogenic Long-Evans rats (LE). In addition, we studied the hypothalamic AVP content in LE rats after HA infusion or exposure to restraint stress. HA (270 nmol) administered intracerebroventricularly (i.c.v.) or 5 min of restraint stress caused a 6- to 7-fold increase in plasma concentrations of ACTH and beta-END in LE rats but only a 2- to 3-fold increase in DI rats (p < 0.01 vs. LE). The basal hypothalamic content of CRH and catecholamines (epinephrine, norepinephrine, and dopamine) was similar in DI and LE rats. The hypothalamic AVP content in LE rats was unaffected by central HA infusion or restraint stress and was undetectable in DI rats. We conclude that inherited lack of AVP impaired the ACTH and beta-END response to central HA administration as well as to restraint stress, suggesting that AVP is important for the mediation of these responses. PMID- 8232763 TI - Regulation of prothymosin alpha mRNA levels in rat pituitary tumor cells. AB - Prothymosin alpha (PTA) mRNA and histone H4 (H4) mRNA levels were studied in various experimental conditions that affected GH1 pituitary tumor cell proliferation. Cell proliferation and progression through the cell cycle was assessed by counting cells, 3H-thymidine incorporation and flow cytometry. PTA mRNA levels were decreased in a time-dependent fashion following serum deprivation; when the cells were induced to grow by serum refeeding, PTA mRNA expression was greatly stimulated. Interestingly, after caprylic acid treatment (2.5 mM for 24 h) that arrested cells in the G0/G1 phase of the cell cycle, PTA mRNA and H4 mRNA levels were almost undetectable; conversely, following caprylic acid withdrawal, PTA mRNA and H4 mRNA expression were greatly stimulated. Furthermore, cells cultured in T3-deprived serum, which was found to decrease GH1 cell proliferation, had low levels of PTA and H4 mRNAs. This effect was reversed by the addition of nanomolar concentrations of T3 to the culture. On the other hand, IGF-1 addition to the culture did not substantially modify PTA mRNA levels. The present data clearly indicate that PTA mRNA expression is tied to the proliferating activity of GH1 cells and, thus, could be used as a marker of the action that various agents have on GH1 cell proliferation. PMID- 8232764 TI - Effect of single and repeated immobilization stress on the heat shock protein 70/90 system of the rat: glucocorticoid-independent, reversible reduction of Hsp90 in the liver and spleen. AB - Heat shock proteins (hsps) and glucocorticoids are produced in response to many common stressors. In addition, hsps interact physically with the intracellular glucocorticoid receptor (GR) and facilitate its activation by the hormone. To study the effect of stress on the hsp70/90 system and the potential association of the system with the hypothalamic-pituitary-adrenal (HPA) axis, we subjected 3 month-old male Harlan-Sprague Dawley rats to single or repeated (once daily for 6 consecutive days) immobilizations, and measured hsp70/90 steady-state levels in a panel of tissues, as well as circulating ACTH and corticosterone concentrations before, during and after immobilization. Single or repeated immobilizations had, as expected, a profound stimulatory effect on the HPA axis but did not influence the steady-state levels of hsp70/90 in any of the gross brain regions (pituitary, hypothalamus, hippocampus or brain cortex) or most peripheral tissues (thymus, adrenal glands, testes) examined. The hsp90 levels, however, were markedly and reversibly decreased in the liver and spleen both by single and repeated immobilizations. The potential inhibitory effect of glucocorticoids on liver and spleen hsp90 was investigated in bilaterally adrenalectomized rats treated with placebo or oral pharmacologic doses of corticosterone for 1 week. Neither adrenalectomy nor corticosterone treatment had an effect on the hsp70/90 system, suggesting that factors other than glucocorticoids mediate the effect of stress on hsp90 concentrations in the liver and spleen. We conclude that acute and chronic stress are associated with a reversible reduction of hsp90 in the liver and spleen, and this appears independent of glucocorticoids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232765 TI - Structure-related effects of CRF and CRF-derived peptides: dissociation of behavioral, endocrine and autonomic activity. AB - Two C-terminal and one N-terminal peptide fragments derived from corticotropin releasing factor (CRF-1-41), CRF-28-41, CRF-34-41, and CRF-1-8, and the CRF receptor antagonist alpha-helical CRF-9-41 (alpha hCRF) were evaluated for their behavioral, endocrine and autonomic nervous effects in rats. To this purpose, three different approaches were used. First, rats were tested in a passive avoidance behavioral (PAB) task after intracerebroventricular (i.c.v.) injection the peptides at five different doses. I.c.v. CRF-1-41 was found to attenuate PAB. The effects of CRF-1-41 on PAB were completely antagonized by i.c.v. pretreatment with alpha hCRF. When given as a sole treatment, the antagonist produced a bimodal effect on PAB. At low doses, alpha hCRF tended to facilitate PAB, while high doses of the antagonist significantly attenuated PAB. Both CRF fragments showed behavioral effects similar to CRF-1-41. Of all peptides tested, CRF-34-41 was found to most attenuate PAB at both retention tests. In a second experiment, the behaviorally most potent fragment CRF-34-41 was compared to CRF-1-41 for its adrenocorticotropic activity after iv injection in pentobarbital-anesthetized rats. Treatment with CRF-1-41, in a dose-related fashion, produced a significant rise in plasma ACTH, whereas CRF-34-41 was without effect. Finally, the effect of i.c.v. injected CRF-1-41 and CRF fragments on heart rate (HR) and gross activity was measured in conscious rats in their home cages during a 60-min period, using a wireless telemetry system. Concomitantly, the occurrence of grooming behavior was recorded. During the first 10 min after i.c.v. treatment, of all peptides tested, CRF-34-41 produced the most marked increase in HR, which remained significant only during the first 30 min of recording. CRF-28-41 induced a non significant transient tachycardia. The parent molecule CRF-1-41 also induced an immediate, significant tachycardia but this effect lasted longer than 60 min. The N-terminal CRF-1-8 remained without effect. No significant effects on gross activity were observed with the two short C-terminal peptides, whereas i.c.v. injected CRF-1-41 induced excessive grooming behavior. A significant grooming response was also recorded in rats given CRF-34-41 fragment, but not in those treated with CRF-28-41 or CRF-1-8.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8232766 TI - Effects of the glucocorticoid antagonist RU 486 on pituitary-adrenal function in patients with anorexia nervosa and healthy volunteers: enhancement of plasma ACTH and cortisol secretion in underweight patients. AB - To further explore whether the hypercortisolism of anorexia nervosa reflects an alteration in the set point for corticotropin-releasing hormone (CRH) secretion or is a manifestation of glucocorticoid resistance, we examined plasma ACTH and cortisol responses to the competitive glucocorticoid antagonist RU 486 (10 mg/kg, p.o. at 8.00 h) versus placebo (PBO) in 7 healthy female volunteers and 8 patients with DSM-III-R anorexia nervosa, all of whom were studied while underweight [64.3 +/- 2.1% average body weight (ABW), mean +/- SE] and 5 of whom were restudied longitudinally following refeeding (> or = 85% ABW, mean 87.4 +/- 0.4% ABW). Blood samples were obtained from 16.00 to 16.30 h and from 4.00 to 8.00 h following dosing. Underweight anorexics were significantly hypercortisolemic by 24 h urinary free cortisol excretion compared with controls (239 +/- 37 vs. 119 +/- 12 nmol/day, p < 0.01). Both controls and underweight anorexics had robust early morning (4.00-8.00 h) plasma cortisol responses to RU 486 (465 +/- 61 and 719 +/- 49 nmol/l) compared with PBO (370 +/- 52 and 451 +/- 31 nmol/l; p < 0.02 and p < 0.01, respectively). The underweight anorexics showed a significant mean early morning plasma ACTH response to RU compared with placebo (3.28 +/- 0.63 vs. 2.01 +/- 0.24 pmol/l, p < 0.05), while the controls showed a trend toward an increase in mean plasma ACTH after RU (3.11 +/- 0.36 pmol/l) compared with PBO (2.31 +/- 0.41 pmol/l, p < 0.13); plasma ACTH means were greater on the RU day than the placebo day at 20 of 25 sampling points (p < 0.001). However, the increment in ACTH on the RU day compared to the placebo day was greater in the underweight anorexics at the first 20 of 25 consecutive time points of the early morning sampling period (p < 0.001). Moreover, underweight anorexics showed a significant plasma ACTH and cortisol response to RU 486 at 16.00-16.30 h (8-8.5 h following administration), while the controls showed no significant response of plasma ACTH or cortisol at this time. When restudied following weight recovery, anorexic patients showed reductions in 24-hour urinary free cortisol excretion (to 191 +/- 40 nmol/day) which were no longer significantly elevated compared with control values.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8232767 TI - Adrenal steroid receptors in the PVN: studies with steroid antagonists in relation to macronutrient intake. AB - These studies tested the impact of steroid receptor antagonists on food intake induced by steroid agonists implanted into the paraventricular nucleus (PVN) and also on feeding that naturally occurs at the onset of the active (dark) period in the rat. Implants of corticosterone (CORT) or the selective type II agonist RU28362 in the PVN stimulated feeding in adrenalectomized (ADX) rats, specifically by enhancing carbohydrate ingestion. This feeding response induced by CORT or RU28362 was blocked by PVN implants of the type II antagonist RU486 but was unaffected by the type I antagonist RU28318. In contrast, the type I agonist aldosterone (ALDO) in the PVN stimulated feeding in both sham and ADX rats by preferentially enhancing fat ingestion, which could be inhibited by the type I antagonist RU28318 but not by the type II antagonist RU486. These results indicate that the feeding elicited by CORT at dark onset is dependent upon the functional integrity of type II glucocorticoid receptors within the PVN, in contrast to the feeding elicited by ALDO which is dependent upon endogenous type I steroid receptor activation within this nucleus. Test results with these antagonists alone in freely feeding animals support a functional role for these PVN steroid receptors in adrenal steroid control of natural food intake. Specifically, blockade of PVN type II receptors with RU486 in intact rats selectively suppressed spontaneous carbohydrate feeding at dark onset, while PVN implants of the type I receptor antagonist RU28318 caused a suppression of spontaneous fat intake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232768 TI - Long-term testosterone or diethylstilbestrol treatment affects gamma-aminobutyric acid and central-type benzodiazepine receptors but not peripheral-type benzodiazepine receptors in the female rat brain. AB - Steroids have often been associated with modulation of the GABAergic system in the central nervous system, mainly in ovariectomized rats. In the present study, the effect of the synthetic estrogen diethylstilbestrol (DES) and testosterone (T) on the density of peripheral and central benzodiazepine (BZ) and gamma aminobutyric acid (GABAA) receptors was evaluated in the frontoparietal cortex and whole cerebellum of female rats during the peripubertal period. The density of peripheral-type BZ receptors was not altered in either of these organs, whether or not treated with DES or T. The density of central BZ and GABAA receptors in either frontoparietal cortex or whole cerebellum was significantly reduced following treatment with DES or T; however, the effect of DES was much more pronounced. The similarity of the effect of T to that of DES may suggest that the effect of T is mediated at least partially by intraovarian biosynthesis of estradiol-17 beta from the exogenously administered T. Collectively, these results may suggest that in female rats during the peripubertal period, sex steroids produce a down-regulatory effect on expression of the brain GABAA/BZ complex, in contrast to their well-established up-regulatory effect in adult ovariectomized rats. PMID- 8232769 TI - Prenatal development of glucocorticoid receptor gene expression and immunoreactivity in the rat brain and pituitary gland: a combined in situ hybridization and immunocytochemical analysis. AB - By means of in situ hybridization and immunocytochemical techniques it has been possible to follow the prenatal development of glucocorticoid receptor (GR) messenger RNA (mRNA) expression and GR immunoreactivity (IR) in the rat brain from embryonic day (E) 15 to 22. A 700-base-pair GR cDNA fragment was used for RNA probe generation. In the immunocytochemical analysis a mouse monoclonal antibody (IgG2a) against the rat liver GR was used in combination with the indirect fluorescence technique or the avidin-biotin immunoperoxidase method. At E15 till E22 a moderate to strong GR mRNA signal was observed within the neuro epithelium from the medulla oblongata to the telencephalon. A moderate to strong labelling was also present within the paraventricular hypothalamic nucleus, the arcuate nucleus, the nucleus raphe magnus, the nucleus raphe obscurus and the locus coeruleus. In these areas a weak to moderate nuclear GR IR developed in nerve cells 1 or 2 days after the appearance of the GR mRNA signal. From E15 the adenohypophysis showed the strongest expression of GR mRNA. At E17 a strong GR IR was especially demonstrated in the nuclei of many pituitary cells, some exhibiting adrenocorticotropin IR. The results open up the possibility that there exist active GR in embryonic life capable of regulating proliferation events within the adenohypophysis and the neuro-epithelia of the brain. This embryonic GR may modulate the development of inter alia neuro-endocrine areas such as the paraventricular and arcuate nuclei and arousal-related areas such as the central 5-hydroxytryptamine and noradrenaline neuronal systems. Provided that this embryonic GR is capable of becoming activated by glucocorticoids in fetal life, it may mediate several neurochemical and behavioural impairments caused by prenatal stress. PMID- 8232770 TI - Effects of neuropeptide Y on gonadotropin and prolactin release in normal, castrated or flutamide-treated male rats. AB - This study was carried out to clarify the central effects of neuropeptide Y (NPY) on gonadotropin and prolactin (PRL) release in the male rat and to evaluate the hypothesis that it may play a role in negative feedback regulation of gonadotropin secretion in the male. NPY (4 micrograms in 2 microliters 0.9% NaCl) or an equal volume of saline was microinjected into the third cerebral ventricle of conscious, unrestrained rats and its effects on plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH) and PRL concentrations, and the gonadotropin response to LH-releasing hormone (LHRH) were measured by radioimmunoassay in noncastrated, castrated (3 days before the experiment) rats or animals treated with flutamide, an androgen receptor blocker (10 mg/kg b.w. per day subcutaneously for 3 days). Blood samples were drawn periodically from a Silastic catheter in the external jugular vein just before and for 120 min after intraventricular injections. At 90 min after injection 25 ng of LHRH were infused intravenously and 3 additional blood samples were withdrawn at 10-min intervals after the injection. Normal male rats responded to NPY with a decrease in plasma LH concentrations at 60 min. Plasma LH was significantly lower at 120 min than values in the saline-injected controls. At that time there was a significant 2 fold augmentation of their response to LHRH. In contrast, NPY elevated plasma FSH significantly versus values in controls at 15 and 30 min after injection, but the FSH release in response to LHRH was not altered.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232771 TI - Feed restriction in prepubertal lambs: effect on puberty onset and on in vivo release of luteinizing-hormone-releasing hormone, neuropeptide Y and beta endorphin from the posterior-lateral median eminence. AB - The exact nature of the interaction between energy balance and reproduction is still elusive. Theoretically, nutrition-related variables must reach the hypothalamic luteinizing-hormone-releasing hormone (LHRH) network and/or its neuronal inputs, to alter plasma luteinizing hormone (LH) and therefore reproductive activity. In an attempt to assess the potential mechanism of such interaction at the median eminence (ME) level, the area of hypophysiotropic LHRH neuronal terminals and release, we used a decreased caloric intake lamb model which delays the onset of puberty. Thus, we determined the in vivo release of neuropeptides, by push-pull cannula (PPC) sampling from the posterior-lateral ME, in feed-restricted (FR) ewe lambs and in full-fed (FF), age-matched, contemporary control animals. Specifically, we assessed: (1) serum LH and ME in vivo release of LHRH, beta-endorphin (beta-END) and neuropeptide Y (NPY); beta-END and NPY are two putative neuronal inputs to LHRH neuronal terminals at the ME, reported to be involved in the control of both reproduction and feed intake; (2) the effect that exogenous infusion of beta-END through the PPC might have on the release of ME LHRH and NPY, and on plasma LH. In contrast to other works, the present results were obtained in lambs with intact ovaries. Furthermore, FR lambs were always compared statistically with FF contemporary paired controls that had attained puberty. Feed restriction decreased ME LHRH release, lowered plasma LH and prevented the onset of puberty. The changes induced by feed restriction in both LHRH and LH release were associated predominantly with decreases in pulse amplitude, rather than alterations in pulse frequency. The decreased LHRH and LH release occurred in the presence of a decreased beta-END but unchanged NPY release from the ME. Exogenous infusion of beta-END into the posterior-lateral ME decreased both LHRH and NPY release from this site and decreased plasma LH. In conclusion, decreased caloric intake lowers LH release and prevents puberty onset by decreasing the amplitude of the LHRH output from the hypothalamic hypophysiotropic network. A compensatory but unsuccessful mechanism for the FR status might be a lower beta-END-inhibitory tone on ME LHRH neuronal terminals. The unchanged release of NPY at this site supports the specificity of the changes induced by feed restriction on LHRH and beta-END in vivo release. PMID- 8232772 TI - Beta-endorphin regulation of luteinizing hormone-releasing hormone release at the median eminence in ewes: immunocytochemical and physiological evidence. AB - Beta-endorphin (beta-END) is an inhibitory factor in the neuroendocrine control of luteinizing hormone (LH) release and thus, presumably also of hypophysiotropic luteinizing hormone-releasing hormone (LHRH) release. In order to address if the median eminence (ME) is a site of beta-END action, we studied its functional role in ewes by assessing: (a) the hypothalamic distribution of beta-END using immunolabeling and by comparing this distribution with our data on the localization of LHRH; (b) the ME in vivo release of LHRH and beta-END during the luteal (day 12) and the follicular (day 15) phases of the estrous cycle; (c) the in vivo release of LHRH from the posterior-lateral ME, as assessed by push-pull cannula (PPC) sampling, before, during, and after infusion of increasing doses of beta-END or naloxone through the PPC, during the follicular phase; and (d) the in vivo release of ME-LHRH and serum LH, before, during, and after infusion of beta END or naloxone in luteal and follicular ewes. In the ewe, beta-END-containing perikarya are located in and around the arcuate nucleus. Their processes are also present in the diagonal band, medial septal nucleus, and medial and lateral hypothalamic areas, including the preoptic region and posterior ME. Perikarya containing LHRH are located in the preoptic area and project also to the ME, providing opportunities for synaptic interactions between beta-END and LHRH containing perikarya and processes at these levels. ME in vivo release of LHRH and beta-END increase from the luteal (low LH/high progesterone, P4) to the follicular phase (high LH/low P4). In follicular ewes, in vivo LHRH and LH release is decreased, in a dose-dependent manner, by beta-END infused through the PPC probe into the posterior-lateral ME. In contrast, infusion of naloxone under similar conditions increases LHRH and LH release, also in a dose-dependent fashion. The inhibitory effect of beta-END on LHRH and LH, as well as the stimulatory effect of naloxone on LHRH and LH, were only marginally apparent in luteal ewes. These results suggest that the ME is a major control site where beta END exerts its influence on hypophysiotropic LHRH release. The strength of this inhibitory effect apparently increases throughout the follicular phase, and might prevent the premature onset of the preovulatory surge of LHRH and LH. PMID- 8232773 TI - Effect of different chronic intermittent stressors and acetyl-l-carnitine on hypothalamic beta-endorphin and GnRH and on plasma testosterone levels in male rats. AB - Chronic stress affects the reproductive function by modifying the neuroendocrine homeostasis. The aim of the present study was to clarify the neuroendocrine and the gonadal changes following chronic intermittent stress in male rats and the action of a neuroactive drug, acetyl-l-carnitine (ALC). The effect of two different stressors, cold water swimming or ether, on central beta-endorphin (beta-EP) and GnRH contents, and on plasma testosterone levels was investigated. In addition, the response to an acute stress in chronically stressed rats, treated or untreated with ALC (10 mg/day/rat p.o.), was evaluated. The stressors were applied twice a day for 10 days, and rats were killed before, during and after the last stress session. Mediobasal hypothalamus (MBH) beta-EP and GnRH contents, and plasma testosterone levels were evaluated by radioimmunoassay. The following results were obtained: (1) both chronic swimming and ether stress caused a decrease in hypothalamic beta-EP contents; (2) MBH GnRH contents increased after chronic swimming stress but not after ether stress; (3) chronic swimming stress induced a twofold decrease in plasma testosterone levels, while no changes were observed after ether stress; (4) the treatment with ALC prevented the decrease in plasma testosterone levels after chronic swimming stress, and (5) acute stress in chronically stressed animals caused an increase in MBH-beta-EP. The present data showed that chronic swimming stress reduces the reproductive capacity and impairs the capacity to respond to the acute stress and that ALC modulates the hormonal changes to physical stress and prevents the antireproductive effect of chronic cold swimming. PMID- 8232774 TI - Experimental evidence for an early commitment of gonadotropin-releasing hormone neurons, with special regard to their origin from the ectoderm of nasal cavity presumptive territory. AB - The origin and the migration of gonadotropin-releasing hormone (GnRH)-producing neurons were studied using the indirect immunoperoxidase method in normal and surgically operated chick embryos. In normal embryos, during early embryonic development, GnRH neurons were located only in the respiratory and the olfactory epithelia. Then, these neurons followed the nearest nerve bundle and occupied, thereafter, the dorsal, medial or ventral part of the olfactory nerve according to the time and area of the olfactory epithelium they emerged from. At the junction with the forebrain, the majority of GnRH neurons passed ventromedially round the olfactory bulb. Therefore, they penetrated through the interhemispheric space and coursed obliquely toward caudal and dorsal telencephalon from where they will be later distributed to reach their adult-like position. In view of the large distribution of these neurons in the nasal region, unilateral surgical ablation either of the whole or of each presumptive territory of nasal structures was performed from 2 to 4 somite stages. As expected, when both olfactory placode and ectoderm of nasal cavity presumptive territories were unilaterally removed, olfactory nerve, nasal structures and GnRH neurons failed to develop in the operated side. After the unilateral removal of the olfactory placode anlage, the distribution pattern of GnRH neurons was not disturbed in the operated as well as in the control side although ipsilateral olfactory structures were greatly reduced. In contrast, when the presumptive ectoderm of nasal cavity was unilaterally removed, GnRH neurons were detected only in the control side where this territory was left intact. Therefore, from early neurogenesis, GnRH neurons seem to be already committed, and they originate from the ectoderm of nasal cavity presumptive territory. PMID- 8232775 TI - Effects of electrical stimulation on muscles of children with Duchenne and Becker muscular dystrophy. AB - Twelve children with progressive muscular dystrophy (10 Duchenne and 2 Becker type) were included in a low-frequency electrical stimulation (LFES) program of the right tibialis anterior (TA) muscle for three months. Muscle strength was estimated by measuring torques in the ankle during short attempts of maximal voluntary isometric contraction (MVIC) in the direction of dorsal flexion of the foot. Muscle fatigue was assessed by the decrease of force during sustained (1 minute) voluntary contraction. The measurements were carried out before the beginning of the stimulation program and immediately after its conclusion. At the end of the stimulation program there were higher torques in 10 out of 12 children in the stimulated leg. The increase in torques in the stimulated leg was statistically significant (p < 0.01). Regarding the fatigue of the stimulated muscle there was no change after the conclusion of stimulation. PMID- 8232776 TI - EEG features of cortical dysplasia in children. AB - Two distinctive electroencephalographic abnormalities, very high amplitude rhythmic activity or prominent fast activity, have been described in children with extensive cortical dysplasia. Cases with cortical dysplasia identified on computerised tomography or magnetic resonance imaging, or a characteristic EEG were selected. One hundred and forty electroencephalograms from 94 cases were reviewed and related to the imaging findings. An EEG with very high amplitude rhythmic activity was found to have high specificity for severe cortical dysplasia but low sensitivity (< 50% cases). Abnormal fast activity was not specific and was seen with very diverse pathologies. The EEG features of most cases with localised cortical dysplasia were very variable. The EEG could be normal even when the cortical dysplasia was extensive. PMID- 8232777 TI - Clinicopathological study of pontosubicular necrosis. AB - Clinicopathological features were examined in 65 neonates and 8 stillborn infants with pontosubicular necrosis (PSN) compared to 57 neonates and 19 stillborn infants without PSN. Twelve out of 65 neonates with PSN had congenital heart disease and 3 out of 65 neonates showed persistent fetal circulation. On neuropathological examination, the frequency of neonates with PSN who also showed karyorrhetic/eosinophilic neurons in other regions of the brain stem, basal ganglia and thalamus was higher than in controls. The results of this study suggest that acute ischemia is an important underlying pathogenetic factor and PSN occurs in the prenatal as well as postnatal period. PMID- 8232778 TI - Exclusion of linkage of genetic focal sharp waves to the HLA region on chromosome 6p in families with benign partial epilepsy with centrotemporal sharp waves. AB - Benign partial epilepsy with centrotemporal sharp waves (benign rolandic epilepsy, BRE) is a common form of idiopathic, localisation-related epilepsy of childhood. The characteristic age-dependent focal sharp wave (fsw) found on the EEG in this disorder segregates as an autosomal dominant trait in families with probands with BRE and acts as a neurobiological marker for the increased risk of developing BRE, other benign partial epilepsies of childhood, and other developmental disorders in these families. One of the genes for idiopathic generalised epilepsy (IGE), designated EJM1, has been mapped in families with probands with juvenile myoclonic epilepsy, by linkage to the HLA region on chromosome 6. As BRE and IGE are benign, idiopathic, age-dependent epilepsies, EJM1 is a candidate locus for the fsw underlying BRE and related disorders. Genetic linkage analysis was undertaken in 11 families with probands with BRE and one or more first degree relatives with fsw, with or without BRE, using a polymorphic DNA marker within the HLA region. Apparently unaffected individuals were classed as affection status unknown. Assuming autosomal dominant inheritance with a penetrance of 0.9 gave a lod score of -2.3 at zero recombination, excluding the candidate gene region around HLA. These observations exclude an important candidate gene for this common disorder, and suggest a fundamental molecular and genetic distinction between the benign partial epilepsies of childhood and the idiopathic generalised epilepsies. PMID- 8232779 TI - Benign childhood epilepsy with centrotemporal spikes and the focal sharp wave trait is not linked to the fragile X region. AB - Benign childhood epilepsy with centrotemporal spikes (BCECS, benign rolandic epilepsy) is a common form of genetically determined localisation-related epilepsy of childhood. The characteristic age-dependent focal sharp wave (fsw) found on the EEG in this disorder segregates as a dominant trait in families with probands with BCECS. Seizures occur in a significant proportion of individuals with the fragile X syndrome in association with EEG abnormalities comparable to those found in BCECS. The possibility of a common genetic basis for these disorders was investigated by linkage analysis. Six pedigrees with probands with BCECS were analysed using a marker locus DXS548, close to the fragile X site, fra (X). Obligate recombinants between DXS548 and the fsw trait were observed in all six families. Assuming X-linked dominant inheritance and penetrance values of 0.4 (male) and 0.1 (female) a negative lod score of -6.823 was obtained at zero recombination and lod scores of -2.0 at 10cM either side of the fra (X) locus. These results exclude an important candidate gene for this common childhood disorder. PMID- 8232780 TI - Reversal of brain atrophy with biotin treatment in biotinidase deficiency. AB - Two children with biotinidase deficiency presented with seizures at 2 months of age. The first child had a fluctuating course with continual developmental progress and cessation of seizures despite symptoms of chronic neurologic dysfunction until he was diagnosed at 17 months. The second child had a progressive course with uncontrolled seizures leading to an unresponsive state until she was diagnosed at 6 1/2 months. Neither child had dermatologic symptoms until shortly before the time of diagnosis. Both children improved markedly with biotin treatment. Serial CT-scan and MRI studies of the brain showed a distinct pattern of changes. Shortly after initial presentation, diffuse low attenuation of the white matter was seen followed by progressive marked cerebral atrophy, which was reversed following biotin treatment. Because this is a reversible condition, clinicians should screen for biotinidase deficiency in all children with symptoms of chronic neurologic dysfunction, especially when radiologic findings of low attenuation of the white matter are followed by cerebral atrophy. PMID- 8232781 TI - Eye movements in children with opsoclonus-polymyoclonus. AB - The anatomical localisation of the abnormality underlying opsoclonus polymyoclonus-the "Dancing Eye Syndrome"-is uncertain and both the brainstem and cerebellum have been implicated. We used electrooculographic recordings to assess the eye movements in five children with this syndrome. Overshoot dysmetria was the consistent feature for the saccades of all five cases. One patient was also assessed after the main symptoms of ataxia and opsoclonus (saccadic oscillations, without an intersaccadic interval) had resolved and the saccades were found to remain hypermetric. Smooth pursuit and optokinetic nystagmus were normal, although the opsoclonus occurred throughout the recording session and were superimposed upon the normal eye movement waveforms. The vestibulo-ocular reflex was normal in four patients, but in one case, the time-constant was significantly prolonged: this has only been reported in experimental lesions of the nodulus in monkeys. The presence of saccadic overshoot dysmetria is highly suggestive of a cerebellar origin for the abnormal eye movements in these children. However, the preservation of normal smooth pursuit and the absence of gaze-paretic, rebound and downbeat nystagmus, implies the sparing of the flocculus and paraflocculus. We propose that the origin of these oculomotor deficits could be the cerebellar fastigial nuclei. PMID- 8232782 TI - Congenital hemiplegia: morphology of cerebral lesions and pathogenetic aspects from MRI. AB - We have analyzed the MRI findings from the brains of 33 children with congenital hemiplegia. Referral of these children to our hospital was either because of neurological problems or a history of complicated birth. According to maturation dependent pathophysiological mechanisms we have classified the lesions into the following five groups: 1. malformations/prenatal encephalo-clastic lesions, 2. periventricular leukomalacia or atrophy, 3. diencephalic lesions, 4. subcortical and cortical lesions, and 5. normal findings. Combination of lesions was not uncommon. The neuroradiologically most prominent and most expanded lesions determined the classification to the different groups. We detected malformations/encephalo-clastic lesions (Group 1) in 5 children; one of these children also presented additional lesions of Groups 2 and 3. Six children displayed periventricular leukomalacia (Group 2), and in one child in combination with diencephalic and subcortical lesions. Ten children exhibited diencephalic lesions (Group 3), in one case combined with periventricular leukomalacia. The MRI of seven children showed subcortical/cortical lesions (Group 4), in four cases extending into diencephalic structures. Two children had a combination of evenly matched periventricular, diencephalic and subcortical/cortical lesions, where it was impossible to define a principal lesion. Three children had normal MRI findings. Significantly, 8 of 33 children had bilateral lesions although presenting with hemiplegia. The large proportion of diencephalic lesions, not described in similar CT studies, and the small number of normal MRI findings show the value of MRI in evaluation of congenital hemiplegia. The ability to correlate, to some extent, neuroradiological findings of damage to developmental stage affords the conclusion that at least a third of the children in our series with congenital hemiplegia suffered prenatal damage. PMID- 8232783 TI - Vigabatrin in the treatment of infantile spasms. AB - The anti-epileptic effect of vigabatrin as a first choice therapy was investigated in 6 children suffering from infantile spasms (IS). All 6 children showed a reduction in seizure frequency. Three children became seizure-free within a period of 2 weeks. Because our results suggest "an all or nothing phenomenon", a period of 2 weeks may be sufficient to evaluate the efficacy of vigabatrin in untreated infants suffering from IS. We suggest to use vigabatrin as a first choice anti-epileptic drug in infants with IS. PMID- 8232784 TI - Improvement of myotonia with carbamazepine in three cases with the Schwartz Jampel syndrome. AB - Carbamazepine was used as an anti-myotonic agent in three cases with the Schwartz Jampel syndrome. Clinical myotonia, blepharospasm and a five-stair climbing test improved significantly within two to four months of the initiation of the medication. In the two cases which were followed three and two years, height shifted from under the 3rd centile to the 5th. Carbamazepine may be a drug of choice in the Schwartz-Jampel syndrome. PMID- 8232785 TI - Intravenous immune globulin is also an efficient therapy of acute Guillain-Barre syndrome in affected children. PMID- 8232786 TI - Enzyme replacement therapy of infantile Gaucher disease. AB - We report our experience from enzyme infusion therapy in a girl with infantile (type 2) Gaucher disease. When treatment was started at 5.5 months of age, she already had severe neurological symptoms. After three months of treatment, the hematological parameters and blood glucosylceramide levels were normalized. The spleen and liver sizes were reduced and the neurological deterioration seemed to have stopped. There was, however, no improvement of her existing neurological symptoms. Her lung function deteriorated because of constant aspirations. Enzyme treatment was stopped after seven months. We cannot recommend enzyme substitution therapy when severe neurological signs have already emerged. PMID- 8232787 TI - Occurrence of Andermann syndrome out of French Canada--agenesis of the corpus callosum with neuronopathy. PMID- 8232788 TI - Multiple binding sites for nicotine receptor antagonists in inhibiting [3H](-) nicotine binding in rat cortex. AB - The displacement of [3H](-)-nicotine from its binding site in rat cerebral cortex by a number of antagonists was investigated. [3H](-)-Nicotine appeared to bind to a single site with a dissociation constant (KD) of 5.5 nM; pancuronium, gallamine and trimetaphan displaced [3H](-)-nicotine with inhibition constants (KI) of 57, 99 and 621 microM, respectively, whereas mecamylamine only displaced 50% of nicotine binding in concentration > 1 mM. For hexamethonium and (+)-tubocurarine the displacement of [3H](-)-nicotine binding appeared to involve two sites; the higher affinity site comprising 30% of the total binding for hexamethonium but 76% of the sites for (+)-tubocurarine. In the presence of mecamylamine (100 microM) the displacement of [3H](-)-nicotine binding by (+)-tubocurarine appeared to involve only a single site with an affinity similar to that for the high affinity site in the absence of mecamylamine whereas that for hexamethonium still involved two sites. It is suggested that (+)-tubocurarine may act at both the binding site for [3H](-)-nicotine per se and also at an allosteric site. The failure of mecamylamine to influence the binding of hexamethonium suggests that there may be more than one allosteric site or that hexamethonium may distinguish between subtypes of nicotine receptors in the cortex. PMID- 8232789 TI - Effects of buprenorphine on motility in chronically morphine treated rats. AB - The effects of buprenorphine (0.01-0.1 mg/kg) on the activity of nondependent and morphine dependent rats (a 20 mg/kg dose for 28 days) were determined. In naive animals buprenorphine exhibited both depressive and stimulatory actions upon the motility of rats, as repeatedly described for morphine. When buprenorphine was administered to rats chronically treated with morphine, the depressive effect disappeared (cross-tolerance to the inhibitory action); on the contrary the excitatory effect was enhanced (cross-sensitization). The results are discussed in terms of cross-sensitization to the excitatory effects in morphine dependent animals as predictor of morphine-like addictive properties in humans. PMID- 8232790 TI - Different densities of 5-HT3 receptors are labeled by [3H]quipazine, [3H]GR 65630 and [3H]granisetron. AB - The binding of three, structurally distinct, 5-hydroxytryptamine3 (5-HT3) receptor radioligands was characterized in rat cerebral cortex, rabbit ileum myenteric plexus and NG-108-15 neuroblastoma cells. The density of sites labeled by the three ligands in rat cortex or in rabbit ileum was markedly different. [3H]Quipazine labeled more sites than [3H]GR 65630 in rat cortex (4.0-fold) and rabbit ileum (1.8-fold), but not in NG-108-15 cells. [3H]Quipazine also labeled a greater density of sites than [3H]granisetron in rat cortex (7-fold) but not in NG-108 cells. [3H]Quipazine binding in rat cortex and rabbit ileum, but not in NG 108-15 cells, was displaced by non-radiolabeled GR 65630 in a manner consistent with an interaction with more than one site. These data indicate that not all 5 HT3 receptor radioligands recognize the same population of 5-HT3 binding sites with equivalent density and further suggest the existence of subtypes of 5-HT3 receptor binding sites in rat cortical and rabbit myenteric plexus preparations. PMID- 8232791 TI - The effect of environmental enrichment on amphetamine-stimulated locomotor activity, dopamine synthesis and dopamine release. AB - In two separate experiments, rats were raised in either an enriched condition (EC) or impoverished condition (IC) from 21 to 60 days of age. Experiment 1 assessed amphetamine-stimulated locomotor activity and in vivo dopamine (DA) synthesis and metabolism in the nucleus accumbens (NA) and striatum (Str). In Experiment 2, amphetamine-stimulated DA release in the NA and Str was assessed in vitro. The results showed that EC rats have lower basal levels of locomotor activity than IC rats. However, in the presence of amphetamine, EC rats showed a greater increase in locomotion over IC when compared to their own controls. Concomitant with this behavioral difference, EC rats also showed an enhanced neurochemical response to amphetamine in vivo. That is, relative to IC rats, amphetamine produced a greater synthesis of DA in the Str of EC rats, as well as a greater metabolism of DA in the NA of EC rats. In the in vitro DA release experiment, EC rats had a lower concentration of tissue DA than IC. However, in contrast to the in vivo experiment, there were no significant differences between EC and IC rats in amphetamine-stimulated release of DA in vitro in either the Str or NA. The failure of amphetamine to produce differential neurochemical effects in EC and IC rats in vitro may be because this experiment eliminated either pharmacokinetic effects or neurochemical differences in brain regions outside the NA and Str. PMID- 8232792 TI - Ammonia-independent modifications of the background EEG signal and paradoxical enhancement of epileptic abnormalities in EEG after acute administration of valproate to epileptic patients. AB - The effects on the background quantitative EEG (power spectral analysis) and concentration of valproate in plasma were studied after single-dose (14.3-33.3 mg/kg) oral administration in 12 epileptic patients with generalized nonconvulsive or partial seizures. An increase of the amplitude of the background EEG (diffuse and preponderant on anterior scalp areas) and a decrease of the 12.5 32.0 Hz relative power (limited to the posterior electrode deviations) were observed; the increase in the EEG total power was paralleled by a definite increment in incidence of epileptic phenomena in the EEG. Both effects proved unrelated to shifts in vigilance or changes in the concentration of ammonia or serum glucose in plasma and confirm previous observations from superimposable study designs. These findings are qualitatively opposite to those observed during long-term treatment at comparable doses and are suggested to reflect a direct CNS action of acute administration of valproate. PMID- 8232793 TI - An antagonist of the metabotropic glutamate receptor prevents LTP in the dentate gyrus of freely moving rats. AB - The effect of the competitive metabotropic glutamate receptor antagonist (RS) alpha-methyl-4-carboxyphenylglycine (MCPG) on long-term potentiation (LTP) in the dentate gyrus was studied in freely moving rats. Intracerebroventricular infusion of MCPG (0.0208 mg) 30 min prior to tetanic stimulation reduced the duration of LTP of the population spike amplitude to 2-3 hr. MCPG itself was without effect on the basal responses evoked by test stimuli. These results provide the first evidence for the requirement of activation of metabotropic glutamate receptors for the induction and maintenance of LTP at perforant path/dentate gyrus synapses in freely moving animals. PMID- 8232794 TI - Pathological heterogeneity of angiographically occult vascular malformations of the brain. AB - There is considerable confusion in the literature regarding the pathological substrates of angiographically occult vascular malformations (AOVMs) of the brain and their clinical significance. We retrospectively reviewed the cases of 34 consecutive patients with AOVMs undergoing surgical excision at a single institution during a 10-year period. Pathological specimens were reexamined, and the lesions were classified according to strict histopathological criteria. There were 21 cavernous malformations, 3 arteriovenous malformations, 3 venous malformations, 2 capillary malformations, and 5 mixed (pathologically heterogeneous) lesions. The initial pathological diagnostic report had been imprecise or had misidentified the lesion type in 18 of the 34 cases (53%), most commonly labeling a cavernous malformation as an arteriovenous malformation or not recognizing mixed features within the same lesion. Clinical presentation (including hemorrhage) and outcome were not significantly different among the various lesion types. Preoperative diagnostic imaging included a variety of modalities that were introduced or evolved during the period of the study and generally suggested a suspected vascular malformation but did not predict pathological subtypes. Acute hematomas in this surgical series made the identification of underlying vascular malformations highly speculative. We conclude that the majority of AOVMs requiring surgical intervention are cavernous malformations, although there was a notable pathological heterogeneity of the remaining lesions. Histopathological subtypes of AOVMs are not associated with unique clinical or radiographic features. PMID- 8232795 TI - Preoperative versus postoperative neuropsychological sequelae of arteriovenous malformations. AB - In a cohort of 14 patients suffering from cerebral arteriovenous malformations (AVM), neuropsychological functioning was examined before and after AVM resection. Improvements after surgery were assumed to be due to enhanced neurocognitive functions associated with the hemisphere ipsilateral to the AVM, and to a lesser extent, with the contralateral hemisphere. Before surgical intervention, the performances of AVM patients were deficient relative to matched normals. Postoperatively, neuropsychological gains were observed particularly in the areas of learning, memory, and higher integrative thought, not only for ipsilateral, but also for contralateral functioning. Contralateral and ipsilateral improvement is consistent with the premise that cerebrovascular steal is lessened; thus, neurosurgical intervention to eliminate arteriovenous shunts was found to result in overall neurobehavioral gains. PMID- 8232796 TI - A prospective study of impairment of cognition and memory and recovery after subarachnoid hemorrhage. AB - In this prospective study, a series of 89 patients with subarachnoid hemorrhage (SAH), most of whom had a "good" neurological outcome, were assessed with a range of tests of memory and cognition as inpatients and at 10 weeks and 12 months after SAH. On tests of verbal cognition and memory, most patients had scores in the normal range 12 months after SAH. However, a significant number of patients still showed impairment on tests of visuospatial construction and memory, mental flexibility, and psychomotor speed at the 12-month assessment. Statistical analyses were carried out for each test score to see whether aneurysm site, location of blood on the admission computed tomographic scan, vasospasm, ischemia, hydrocephalus, grades at admission to and at discharge from hospital, and Glasgow Outcome Scale score at follow-up were associated with test scores. Aneurysm site was not shown to be associated with performance on any test at any time, and the other complications of SAH had only minimal predictive value. The grade at discharge proved to be the best predictor of impairment of cognition and memory at both follow-up assessments. Older subjects did not recover to the same extent as younger subjects by the 12-month assessment. The authors conclude that the diffuse effects of SAH are more important than focal neuropathology in relation to cognitive impairment in this group of patients. PMID- 8232797 TI - Modifications based on computed tomographic imaging in planning the radiosurgical treatment of arteriovenous malformations. AB - Early in the course of treating arteriovenous malformations with radiosurgery, it was noted that the nidus isocenter and diameter, as identified by stereotactic angiography, often differed from that identified by stereotactic enhanced computed tomography (CT). To assess the sources of discrepancy between the arteriographic and CT representations of the nidus, dosimetry (from treatment records stored on an optical disk) was reviewed in 81 consecutive cases. In 44 cases, the isocenters differed by an average of 3.6 mm and the collimator size differed. Fourteen nidi were larger on CT (average, 2.6 mm), and 30 were smaller on CT (average, 4.0 mm). Overall, the angiographic and the CT nidus differed in 75% of the cases reviewed. Sources of error in the angiographic nidus determination included overlapping vessels, bony structures, fine filamentous arterioles, and irregular shapes. PMID- 8232798 TI - Preservation of cranial nerve function after radiosurgery for nonacoustic schwannomas. AB - Microsurgical resection is the primary management approach for patients with intracranial schwannomas. Recent studies have demonstrated that stereotactic radiosurgery is an effective therapeutic modality for patients with acoustic schwannomas. To define the role of radiosurgery in the management of patients with nonacoustic schwannomas, we reviewed the results of gamma unit stereotactic radiosurgery in six patients with trigeminal and five patients with jugular foramen region schwannomas. No patient with a trigeminal schwannoma demonstrated tumor growth during a mean follow-up of 21 months (range, 7-35 mo), whereas one patient with a jugular foramen region schwannoma had an increase in tumor size 7 months after radiosurgery. No new cranial nerve or brain stem deficits were noted in either patient group after radiosurgery. In this early experience, radiosurgery proved an effective primary or adjuvant technique for selected patients with schwannomas of the trigeminal, glossopharyngeal, or vagus nerves. Using our described method, the safety of radiosurgery was demonstrated on the brain stem, regional cranial nerves, and especially those cranial nerves intimately associated with the tumor. PMID- 8232799 TI - A retrospective analysis of pituitary apoplexy. AB - Thirty-seven patients with pituitary apoplexy were analyzed with an emphasis on clinical presentation and visual outcome. Their mean age was 56.6 years, with a male to female ratio of 2:1. Presenting symptoms included headache (95%), vomiting (69%), ocular paresis (78%), and reduction in visual fields (64%) or acuities (52%). Computed tomographic scanning correctly identified pituitary hemorrhage in only 46% of those scanned. Thirty-six patients underwent transsphenoidal decompression. By immunostaining criteria, null-cell adenomas were the most frequent tumor type (50%). Long-term steroid or thyroid hormone replacement therapy was necessary in 82% and 89% of patients, respectively. Long term desmopressin therapy was required in 11%, and 64% of the male patients required testosterone replacement therapy. Surgery resulted in improvement in visual acuity deficits in 88%, visual field deficits in 95%, and ocular paresis in 100%. Analysis of the degree of improvement in preoperative visual deficits with the timing of the surgery demonstrated that those who underwent surgery within a week of apoplexy had significant recovery in their visual acuities. In the stable, conscious patient with residual vision in each eye, surgical decompression should be performed as soon as possible, because delays beyond 1 week may retard the return of visual function. PMID- 8232800 TI - Pituitary adenomas with invasion of the cavernous sinus space: a magnetic resonance imaging classification compared with surgical findings. AB - We present 25 pituitary adenomas that were confirmed surgically to have invaded the cavernous sinus space. The surgical results are compared with the preoperative magnetic resonance imaging findings. For comparable radiological criteria, we classified parasellar growth into five grades. This proposed classification is based on coronal sections of unenhanced and gadolinium diethylene-triamine-pentaacetic acid enhanced magnetic resonance imaging scans, with the readily detectable internal carotid artery serving as the radiological landmark. The anatomical, radiological, and surgical conditions of each grade are considered. Grades 0, 1, 2, and 3 are distinguished from each other by a medial tangent, the intercarotid line--through the cross-sectional centers--and a lateral tangent on the intra- and supracavernous internal carotid arteries. Grade 0 represents the normal condition, and Grade 4 corresponds to the total encasement of the intracavernous carotid artery. According to this classification, surgically proven invasion of the cavernous sinus space was present in all Grade 4 and Grade 3 cases and in all but one of the Grade 2 cases; no invasion was present in Grade 0 and Grade 1 cases. Therefore, the critical area where invasion of the cavernous sinus space becomes very likely and can be proven surgically is located between the intercarotid line and the lateral tangent, which is represented by our Grade 2. We also measured tumor growth rates, using the monoclonal antibody KI-67, which shows a statistically higher proliferation rate (P < 0.001) in adenomas with surgically observed invasion into the cavernous sinus space, as compared with noninvasive adenomas. PMID- 8232801 TI - Low-grade astrocytomas may arise from different astrocyte lineages. AB - The management of low-grade astrocytomas remains a challenge. Although the majority of these tumors have common histological features, they may have very different clinical manifestations and rates of proliferation. Because low-grade astrocytomas are composed of relatively well-differentiated neoplastic cells that closely resemble the astrocytic phenotype, it is possible that some of these lesions express antigens that characterize astrocyte lineages. The authors performed an immunohistochemical analysis of 20 low-grade astrocytomas with A2B5, a monoclonal antibody to a ganglioside found in early postnatal Type 2 (fibrillary) astrocytes, but absent in Type 1 (protoplasmic) astrocytes, and anti glial fibrillary acidic protein to determine whether the expression of these antigens could be used to determine the histogenesis of these tumors. These findings were compared with the clinical and imaging features of these tumors. The percentages of cells positive for A2B5 and glial fibrillary acidic protein was strongly correlated with the location of the tumor within the cortex or white matter and with the length of preoperative symptoms. Tumors based in the cortex contained significantly fewer A2B5-positive and glial fibrillary acidic protein positive cells than white matter tumors. In addition, lesions that caused a relatively short period of preoperative symptoms (< 1 year) had significantly more A2B5-positive and glial fibrillary acidic protein-positive cells than lesions responsible for a long preoperative history (mean, 12.9 years). These findings suggest that slow-growing, cortically based low-grade astrocytomas have a phenotype consistent with the Type 1 (protoplasmic) astrocyte lineage, while white matter low-grade astrocytomas express antigens consistent with the Type 2 (fibrillary) astrocyte lineage. PMID- 8232802 TI - The trigeminal evoked potential: Part I. Long-latency responses in awake or anesthetized subjects. AB - Trigeminal evoked potentials have been elicited with differing methodologies that render responses almost as numerous as the number of laboratories reporting them. Forty-six trigeminal nerves were studied to investigate trigeminal evoked potentials in 20 awake, normal subjects and 20 patients undergoing surgery with general anesthesia. A comparison was made between different methods of surface stimulation and subdermal stimulation. It was concluded that these techniques had no practical use in intraoperative monitoring. PMID- 8232803 TI - The trigeminal evoked potential: Part II. Intraoperative recording of short latency responses. AB - Short-latency (< 10 msec) trigeminal evoked potentials were studied intraoperatively in 31 patients under general anesthesia. Stimulation was performed peripherally in the face as well as centrally at the trigeminal nerve root entry zone. Recordings were made directly from the trigeminal root entry zone and far field from the scalp. Two principal negative deflections were noted, N1.6 and N2.6; and two waves that were less constantly identified were N.9 and N4 6. The latencies of these responses necessitated an extra-axial origin. It was concluded that this technique provided the ability to monitor the integrity of the extra-axial trigeminal nerve, but not the central trigeminal circuitry. PMID- 8232804 TI - Fetal tissue grafting into the central nervous system: yesterday, today, and tomorrow. PMID- 8232805 TI - Endothelium-derived vasoactive factors and regulation of the cerebral circulation. AB - Vasoactive factors produced and released by endothelium exert a powerful influence on vascular tone in the cerebral circulation. Endothelium-derived relaxing factor (EDRF), which has been identified as nitric oxide (NO) or an NO containing compound, is produced under basal conditions in cerebral blood vessels. EDRF mediates endothelium-dependent relaxation in response to a number of stimuli in the cerebral circulation. The influence of NO on the cerebral circulation appears to be particularly important and complex because both neurons and glia, in addition to endothelium, produce NO in response to some stimuli. Neuronally derived NO may mediate local vasodilation in response to increased neuronal activity. In addition to EDRF, cerebral endothelium may produce other relaxing factors, including prostacyclin, endothelium-derived hyperpolarizing factor, and oxygen-derived free radicals. Several pathophysiological conditions are associated with impaired endothelium-dependent responses that may involve the decreased production of EDRF and release of endothelium-derived contracting factors, such as the cyclooxygenase products of arachidonic acid and endothelin. The release of endothelin, an extremely potent and long-lasting vasoconstrictor peptide, may contribute to vasospasm after subarachnoid hemorrhage. PMID- 8232806 TI - A routine stereotactic procedure in 2003. PMID- 8232807 TI - Architecture and functional design of advanced neurosurgical operating environments. AB - Although modern operative neurosurgery is a complex technical undertaking requiring an amalgam of technologies and instrumentations, few reported efforts have dealt with the definition and development of suitable and optimal dedicated operating environments. This report presents the first detailed description of a dedicated, self-contained neurosurgical operating suite incorporating major surgical instrumentation and visualization technologies to provide an "idealized" environment for stereotactic, microscopic, and microstereotactic procedures. Advanced computer technology for visualization to augment, simulate, document, and facilitate all aspects of neurosurgery is described. The architectural and functional design of the operating suite is itself an integral surgical instrument as well as a laboratory for development of new dimensions of neurosurgery. PMID- 8232808 TI - Use of a frameless, armless stereotactic wand for brain tumor localization with two-dimensional and three-dimensional neuroimaging. AB - Preliminary experience with a frameless, armless stereotactic localization system in brain tumor surgery is presented. The localizing wand emits ultrasonic pulses that are detected by a table-mounted array of microphones--with triangulation of the emitter positions. The wand tip and trajectory are determined by proprietary computer software. Real-time display of this information is presented in multiple, two-dimensional or three-dimensional displays. Forty-eight patients underwent 52 craniotomies for brain tumors. The wand was used to assist in placing a minimal craniotomy in 48 cases, to determine the tumor/brain interface in 27 cases, to localize subcortical tumors in 14 cases, and to correlate the physiological mapping with the surface anatomy in 5 cases. In 12 instances, the wand was used in conjunction with frame stereotaxy and found to be comparable or superior. Triplanar (coronal, sagittal, transverse) two-dimensional images provided sufficient information for the detection of tumor boundaries but proved difficult to use to access a subcortical lesion; two-dimensional or three dimensional images along the localization axis were more helpful. Frameless stereotaxy with this sonic wand system proved to be a useful adjunct to open tumor biopsy or resection. PMID- 8232809 TI - Autocrine growth regulation in neuroectodermal tumors as detected with oligodeoxynucleotide antisense molecules. AB - The cell lines of three neuroectodermal tumors, two glioblastomas (HTZ-146, HTZ 17) and one melanoma (HTZ-19) were established and screened for the expression of growth factors by northern blotting and immunochemical methods. All three tumors were positive for platelet-derived growth factor- (PDGF-) A-, -B-chain, and basic fibroblast growth factor (bFGF) messenger ribonucleic acids. Cultured cells as well as original tumor material were also positive for PDGF-AA-, PDGF-BB, and bFGF protein, as shown by immunochemistry. To investigate the possible pathophysiological role of PDGF and bFGF, antisense technology was employed with chemically modified nuclease-stable 14-mer phosphorothioate oligodeoxynucleotides. Proliferation of all three tumors was reduced to a different extent with antisense phosphorothioate oligodeoxynucleotides in vitro, targeted against PDGF-A-chain-, -B-chain-, and -bFGF-messenger ribonucleic acid. These data indicate autocrine stimulatory loops for PDGF and bFGF, which may be blocked, may have different relevance in neuroectodermal tumors in vitro, and may have conceivable future therapeutic implications. PMID- 8232810 TI - Comparison of intrathecal administration of urokinase and tissue plasminogen activator on subarachnoid clot and chronic vasospasm in a primate model. AB - Safety and efficacy of the thrombolytic agent urokinase (URO) in the elimination of subarachnoid clot and prevention of chronic vasospasm was compared with tissue type plasminogen activator (rt-PA) in a blind, randomized placebo-controlled trial. Twenty monkeys were randomly assigned to one of five groups of four. Each group underwent baseline cerebral angiography followed by bilateral craniectomy and experimental subarachnoid hemorrhage. An Ommaya reservoir was inserted on the right side with its catheter placed into the ipsilateral subarachnoid space. Twenty-four hours later, depending upon group assignment, the animals received 100,000 IU URO, 200,000 IU URO, 1 mg rt-PA, 2 mg rt-PA, or the equivalent volume of normal saline (control group). On Day 7, angiography was repeated and the animals were killed. One animal died as a result of complications during the baseline angiography, presumably due to blood loss and prolonged anesthesia, and a replacement animal was obtained. No animals demonstrated any delayed neurological deficits. The study demonstrated that a single intracisternal bolus injection of rt-PA, 2.0 mg in 2 ml sterile water, or URO, 200,000 IU in 2 ml sterile water, 24 hours after induction of experimental subarachnoid hemorrhage in primates, was equally effective in thrombolysing ipsilateral clot, but neither dosage prevented angiographic vasospasm. Vasospasm occurred bilaterally in all groups. Whereas gross subarachnoid clot was found bilaterally in all animals in the placebo group and both smaller-dose URO and rt-PA groups, right-sided subarachnoid clot was virtually absent and left-sided clot reduced in both higher dose URO and rt-PA groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232811 TI - Immunoblotting of contractile and cytoskeletal proteins of canine basilar artery in vasospasm. AB - Vasospasm was produced in the canine basilar arteries by a two-hemorrhage method, and voltage- and receptor-dependent contractions of the normal canine basilar arteries were induced by local applications of potassium chloride (KCI) and serotonin, respectively, after transclival exposure. Actin, myosin, desmin, filamin, talin, vinculin, and alpha-actinin in the basilar artery were studied by immunoblotting. The immunoblots showed a decrease or loss in immunoreactivity of some native proteins and generation of protein fragments, smaller in size than native proteins, in spastic, KCI, and serotonin groups, indicating a proteolytic degradation. In the spastic group on Day 2, actin, desmin, and filamin were usually degraded slightly; myosin moderately; and talin and alpha-actinin substantially. Vinculin and metavinculin remained intact. In the spastic group on Day 7, actin and desmin were usually decomposed slightly; myosin, filamin, and vinculin substantially; and talin, metavinculin, and alpha-actinin markedly. In the KCI and serotonin groups, slight degradation was usually observed in filamin, often in alpha-actinin, and occasionally in actin, whereas desmin, vinculin, and metavinculin were not degraded. In addition, myosin was usually degraded moderately in the KCI group and slightly in the serotonin group, and talin was generally decomposed slightly in the KCI group and moderately in the serotonin group. The degraded fragments, although variable in number and immunoreactivity, were similar in size in the three groups. We suggest that the intracellular devices responsible for contraction of the basilar arteries are degraded more severely in the spastic group than in the KCI or serotonin group, probably by similar proteolytic mechanism and progressively with the passage of time after subarachnoid hemorrhage in vasospasm. PMID- 8232812 TI - Changes in the cerebrovascular effects of endothelin-1 and nicardipine after experimental subarachnoid hemorrhage. AB - The role of endothelium-related factors in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH) has gained interest since the discovery of endothelin-1 (ET-1). We have examined, before and after SAH, the responsiveness of the cerebrovascular bed of the goat to ET-1, the sources of Ca2+ in ET-1 induced responses, and the ability of the Ca2+ entry blocker nicardipine to counteract them. Before SAH, injection of ET-1 into the cerebral circulation increased cerebrovascular resistance, thereby producing dose-dependent reductions in cerebral blood flow (CBF), which were prevented by nicardipine. In isolated middle cerebral arteries, ET-1 induced concentration-dependent contractions, which were equally inhibited in Ca(2+)-free medium (without or with ethylene glycol tetraacetic acid) and by the Ca2+ entry blocker nicardipine. On the third day after SAH, CBF was reduced by 28% and cerebrovascular resistance increased by 39%. At the same time, both ET-1-induced reductions in CBF and the constricting effects of ET-1 in vitro were enhanced. The ability of nicardipine to increase CBF and to inhibit the effects of ET-1 was impaired as a result of reduced dependence of cerebral arteries on extracellular Ca2+. On the seventh day after SAH, CBF and cerebrovascular resistance returned to control values, and effects of ET-1 became normal. It is suggested that the hyperreactivity to ET-1 of the cerebrovascular bed induced by SAH could have a role in the development of vasospasm, which could reduce the vascular effects of Ca2+ entry blockers after SAH. PMID- 8232813 TI - Procollagen types I and III and transforming growth factor-beta gene expression in the arterial wall after exposure to periarterial blood. AB - The stiffening and thickening of the arterial wall after subarachnoid hemorrhage may reflect increased connective tissue. The purpose of this study was to examine the nature of collagen synthesis in response to periarterial blood. Rat femoral arteries were exposed to periarterial blood for varying lengths of time (control, 1, 3, 7, and 14 d). Dot-blot analysis of total ribonucleic acid extracted from the arteries (n = 10 to 15 animals each) demonstrated that the expression of procollagen Types I and III messenger ribonucleic acid increased at 7 (threefold) and 14 days. The expression of transforming growth factor-beta (TGF-beta), an important regulator of collagen synthesis, was markedly increased by 3 days (threefold), followed by a gradual decline. There were marked differences in procollagen Types I and III and TGF-beta gene expression between arteries exposed to blood and sham-operated arteries for a period of 7 days (n = 25 animals). Northern blot analysis of total ribonucleic acid extracted from cultured vascular smooth muscle cells showed that the treatment with a higher concentration of serum for 48 hours increased the expression of procollagen Types I and III and TGF-beta, whereas exposure to oxyhemoglobin did not. After exposure to periarterial blood, arterial walls show increased synthesis of procollagen Types I and III, perhaps a response to the increased secretion of TGF-beta, which in turn could be the result of exposure to serum factors. PMID- 8232814 TI - Vein graft replacement of the middle cerebral artery after unsuccessful embolectomy: case report. AB - A forty-one-year-old man with a cavernous hemangioma of the right cavernous sinus underwent a preoperative cerebral angiogram and a balloon occlusion test of the internal carotid artery. During the operation to remove the cavernous sinus lesion, the ipsilateral electroencephalogram was found to be abnormal. An embolic occlusion of the M2 and M3 segments of the middle cerebral artery (MCA) was discovered. A platelet and thromboembolus was removed via multiple incisions, and flow was restored. The cavernous sinus lesion was removed uneventfully. At the end of the operation, the MCA was found to be reclotted. Flow was eventually restored by replacing the M2 segment of the MCA with a 2-cm saphenous vein graft. The patient recovered without any deficits of brain function and with transient deficits of Cranial Nerves III and VI. Computed tomography revealed infarcts in the temporal and parietal areas. When MCA embolectomy is unsuccessful, vein graft replacement should be considered to restore flow and to avoid major neurological deficits. PMID- 8232815 TI - Hemangiopericytoma of the temporal bone presenting as a retroauricular mass. AB - An unusual case of a hemangiopericytoma arising from the temporal bone is presented. The patient was noted to have a postauricular mass and was neurologically asymptomatic. A preoperative magnetic resonance image and an angiogram revealed the tumor to be highly vascular. Preoperative embolization facilitated the surgical removal of the tumor by rendering it avascular. Current therapy consists of radical resection of the tumor with postoperative radiation therapy. Patients must be monitored carefully for local recurrence and systemic metastasis. PMID- 8232816 TI - A pathological study of intracranial posterior circulation dissecting aneurysms with subarachnoid hemorrhage: report of three autopsied cases and review of the literature. AB - Three autopsied cases of dissecting aneurysms with spontaneous subarachnoid hemorrhage of the intracranial posterior circulation are reported, and the literature is reviewed to investigate the pathological characteristics and pathogenesis of this lesion. The location and pathological features of the aneurysms were different in each of the cases presented. Subadventitial hemorrhage associated with the subarachnoid hemorrhage, multiple noncontiguous intramural hemorrhages, and new vessels in and around the arterial wall were noteworthy findings in our series. All 20 reported autopsied cases of the vertebrobasilar artery have a dissection between the media and adventitia with a rupture site in the thin adventitia. Seventeen of them had disruption of the entire arterial wall, but the remaining three cases had no apparent luminal connection. The clinicopathological features of this disease are various. On the basis of the pathological investigations of these reported cases, the pathogenesis of this lesion is discussed. Intramural hemorrhage associated with dissection without luminal connection should not be disregarded as a causative factor. PMID- 8232817 TI - Dissecting aneurysm of the posterior inferior cerebellar artery: case report. AB - A patient with a dissecting aneurysm of a posterior inferior cerebellar artery who presented with Wallenberg's syndrome is reported. A 31-year-old man suddenly experienced an occipital headache, vertigo, and vomiting, followed by dysphagia. A neurological examination revealed partial Wallenberg's syndrome. Vertebral angiography revealed aneurysmal dilatation at the origin of the left posterior inferior cerebellar artery, with distal luminal narrowing. T1-weighted magnetic resonance imaging demonstrated an area of high-signal intensity, indicating an intramural hemorrhage in the arterial wall of the narrowed lumen. The dissecting aneurysm with a typical intramural hematoma of the posterior inferior cerebellar artery was entrapped with clips after an anastomosis of the left occipital artery to the distal posterior inferior cerebellar artery. The diagnosis and the treatment of dissecting aneurysms of the posterior inferior cerebellar artery are discussed. PMID- 8232818 TI - Extracranial aneurysm of the posterior inferior cerebellar artery: case report. AB - The authors report the unusual case of an aneurysm arising from the extracranial portion of the left posterior inferior cerebellar artery (PICA). The patient was a 39-year-old woman who developed a subarachnoid hemorrhage. The hemorrhage was in the basal, ambient, and Sylvian cisterns, but not in the ventricles. There were no focal neurological deficits, and the clinical grading was 1. On cerebral angiograms, the PICA was found to originate from the vertebral artery as low as 17.9 mm below the foramen magnum and to enter into the spinal canal through the intervertebral foramina of C1-C2. The artery went up to the posterior fossa and had only the cranial loop with an absence of the caudal loop. The saccular aneurysm hung on the initial segment of the PICA and was located at the C1 level. The patient developed motor aphasia and hemiparesis on the right side 15 days after onset, and an infarction was disclosed in the boundary zones of the left parietal lobe on a computed tomographic scan. The aneurysm was successfully obliterated by a neck clipping 2 months later, and the patient was discharged in good condition. The literature discussing PICA aneurysms in the extracranial, as well as distal, portion is reviewed. PMID- 8232819 TI - Improvement of cerebral blood flow and clinical symptoms associated with embolization of a large arteriovenous malformation: case report. AB - We report a patient with an unruptured, large arteriovenous malformation that was treated by staged, superselective embolization with liquid agents and by an investigation of the hemodynamic changes accompanying embolization. A 29-year-old man presented with headache and left upper quadrantanopsia. A neuroradiological study revealed a large right temporo-occipital arteriovenous malformation, and angiography disclosed poor filling of the adjacent vessels. In the venous phase, marked cortical reflux, suggesting venous hypertension, was also observed. Single photon emission computed tomography scanning with N-isopropyl-p-iodine-123- iodoamphetamine disclosed a low-perfusion area in the ipsilateral occipital and temporal lobes. After embolization, cerebral blood flow and the clinical symptoms attributed to ischemia improved. A follow-up study 1 year later demonstrated that the patient's improvement was stable. Single photon emission computed tomography confirmed that embolization achieves an improved cerebral blood flow. PMID- 8232820 TI - Civilian gunshot wounds to the head. PMID- 8232821 TI - Clinical studies of de novo aneurysms. PMID- 8232822 TI - Surgical treatment of moyamoya disease: operative technique for encephalo-dura arterio-myosangiosis. PMID- 8232823 TI - Resection of intraventricular tumors via a computer-assisted volumetric stereotactic approach. PMID- 8232824 TI - Pressure autoregulation is intact after arteriovenous malformation resection. PMID- 8232825 TI - Contribution of calcium ions to the generation of epileptic activity and antiepileptic calcium antagonism. AB - With epileptic activity, neurons show paroxysmal depolarization shifts (PDSs) corresponding to epileptic field potentials (EFPs) generated by the neuronal population. Experimental results indicate that calcium and calcium-dependent currents participate in the generation of these events. Consequently neuronal PDS/EFP were depressed by organic calcium channel blockers. This justifies the hope that calcium channel blockers might be useful in the treatment of human epilepsies. PMID- 8232826 TI - Use of antiepileptic drugs in epileptology with respect to psychiatry. AB - Psychiatric problems in epileptology may result from complications of epilepsies, may parallel manifestations of a common underlying pathology or may be independent of the seizure disorder. The following aspects are described: (1) toxic psychiatric effects of antiepileptic drugs (AEDs); (2) idiosyncratic reactions with psychiatric symptomatology; (3) psychiatric complications at seizure control; (4) AED withdrawal syndromes, and (5) psychotropic actions of AEDs. PMID- 8232827 TI - New developments in the use of anticonvulsants as mood stabilizers. AB - There is increasing recognition that lithium is inadequate in the treatment of up to 50% of bipolar patients. In addition to subgroups that are nonresponsive from the outset, loss of efficacy (tolerance) and discontinuation-induced refractoriness have recently been observed. The anticonvulsants carbamazepine and valproate are effective alternative or adjunctive treatments, but tolerance can also occur during their long-term prophylactic use. New treatment algorithms for this loss of efficacy, including combination therapies, require further systematic study. Preliminary data suggesting that some patients with extremely rapid and chaotic mood fluctuations may respond to the L-type calcium channel blocker nimodipine are presented, and the theoretical implications discussed. PMID- 8232828 TI - Effects of carbamazepine and lithium on affective disorders. AB - Characteristics of the responders and nonresponders to prophylactic treatment with carbamazepine (CBZ) or lithium (Li) were investigated in a retrospective study of 215 bipolar disorder patients who had received more than 2 years of CBZ or Li treatment in Japan. Patients of mania-dominant type, with isolated occurrence of manic and depressive episodes, with onset of the illness below 30 years of age, and with atypical symptomatology, tended to respond favorably to CBZ. Rapid cyclers and patients with a past history of rapid cycling and with a continuous recurrence of episodes responded unfavorably to CBZ. Patients of mania dominant type also showed a favorable response to Li, while patients with a continuous-type course, with rapid cycling, and a past history of rapid cycling, as with CBZ, tended to respond unfavorably to Li. PMID- 8232829 TI - Valproate as a loading treatment in acute mania. AB - Evidence from earlier studies indicates that the antimanic action of valproate becomes most apparent within 1-4 days of achieving serum concentrations 50 micrograms/ml, that valproate can be orally loaded with achievement of therapeutic serum concentrations within the first several days of treatment, there is a rapid onset of response, and minimal side effects. To provide further data on the safety and efficacy of valproate oral loading in the treatment of acute mania, we evaluated 13 consecutive patients with acute manic syndromes who received valproate initiated at a dosage of 20 mg/kg/day. In most cases, valproate was added to other psychotropics. All 13 patients received at least 5 full days of valproate maintained at or above 20 mg/kg/day, and valproate serum concentrations were 50 micrograms/ml (mean +/- SD = 88 + 25) by the second or third day of treatment. Ten (77%) patients displayed a moderate or marked response. Side effects were infrequent and minor. Consistent with our earlier study, these findings suggest that valproate can be safely administered via a loading dose of 20 mg/kg/day to patients with acute mania, including those on other psychotropics, and that it may produce a rapid response with minimal side effects. PMID- 8232830 TI - Antiepileptic drugs for the treatment of panic disorder. AB - Evidence from preclinical studies, preliminary clinical reports, pharmacologic challenge studies and a small number of controlled trials suggests that several antiepileptic agents--valproate, carbamazepine, and clonazepam--may have therapeutic effects in the treatment of patients with panic disorder. We review the theoretical basis and available clinical data supporting the use of these agents in panic disorder. PMID- 8232831 TI - The role of carbamazepine in the prophylaxis of unipolar depression. AB - The prophylaxis of unipolar depression is still a controversial subject. Some authors prefer lithium, others maintenance treatment with antidepressants. The role of carbamazepine remains unclear. Few patients have been described in the literature, and most are lithium nonresponders or rapid cyclers. In an open-label naturalistic study, 15 patients suffering from unipolar depression (DSM-III, 296.2, 296.3) and receiving long-term prophylaxis with carbamazepine were followed for 5 years. Four had been pretreated with lithium without satisfactory effects, 11 were prophylaxis naive. The mean time span patients received carbamazepine was 49.5 months. 73% of the patients gained substantial profit from carbamazepine. Side effects were infrequent. PMID- 8232832 TI - The action of mood-stabilizers in affective disorders: an integrative view as a challenge. AB - One of the main challenges in the psychopharmacology of affective disorders is to explain the partial similarity of the profiles of action of the different mood stabilizers lithium, carbamazepine, and valproate, and possibly also calcium antagonists like verapamil and nimodipine. Elucidation of this problem requires understanding the basic neurobiological mechanisms underlying the pathogenesis of affective disorders and would also imply elucidating the modes of action of the different compounds. However, we are far from reaching such an understanding. PMID- 8232833 TI - The effect of carbamazepine on endocrine and sleep EEG variables in a patient with 48-hour rapid cycling, and healthy controls. AB - Carbamazepine treatment of a patient with 48-hour rapid cycling led to a dampening of mood cycling, and prolonged rapid eye movement (REM) sleep latency. No effect on central alpha-receptors as measured by growth hormone (GH) secretion after clonidine stimulation or on spontaneous 48-hour GH secretion was observed. In 12 healthy subjects given 400 mg carbamazepine daily for a period of 5 days, improved sleep continuity and increased slow-wave sleep occurred with treatment. REM sleep percentage and REM latency remained uninfluenced, whereas REM density decreased. GH secretion after clonidine stimulation was not altered. Data from the single-case longitudinal study emphasize that carbamazepine is effective in treating rapid-cycling affective psychosis. Furthermore, neuroendocrine and sleep EEG data from the study in healthy subjects indicate a different profile of action for carbamazepine compared to most other antidepressants or antimanic drugs. PMID- 8232834 TI - Calcium-antagonistic effects of carbamazepine in epilepsies and affective psychoses. AB - Carbamazepine (CBZ) is known to have beneficial effects in the treatment of epilepsies and in the prophylaxis of affective disorders. Since increased transmembrane calcium fluxes and intracellular calcium concentrations play a key role in the generation of epilepsies and possibly also in the development of these psychiatric disorders the effects of CBZ on epileptic discharges (elicited by caffeine, penicillin and low Mg2+) in CA3 neurons of hippocampal slices were compared with those of the organic calcium antagonist verapamil and found to be almost the same. PMID- 8232835 TI - Effects of carbamazepine on action potentials and calcium currents in rat spinal ganglion cells in vitro. AB - The effects of carbamazepine (CBZ) on action potentials and calcium currents in cultured rat sensory spinal ganglion cells were investigated. CBZ was found to reversibly suppress the calcium-dependent components of action potentials and to reduce the amplitude of the after-hyperpolarizations, while the rising phase and the peak amplitude were hardly changed. Furthermore, CBZ caused a marked reduction in the calcium currents, which in some cells was reversible. The present findings confirm that CBZ has calcium-antagonistic properties. PMID- 8232836 TI - Increased sensitivity to agonist stimulation of the Ca2+ response in neutrophils of manic-depressive patients: effect of lithium therapy. AB - The agonist-stimulated increase of intracellular free-Ca2+ concentration, an indicator of the sensitivity of the inositol phospholipid second-messenger generating system, was measured in neutrophils from patients with manic depressive disorder, and controls. Dose-response curves of the calcium response were determined by measuring the fluorescence of neutrophils loaded with fura-2 and stimulated with various concentrations of the chemotactic tripeptide formylmethionylleucylphenylalanine. EC50 values were obtained for 14 medication free patients (5 acutely depressive, and 9 symptom free remitted patients with a history of manic-depression or recurrent major depression), 9 lithium-treated, euthymic manic-depressive patients and 10 drug-free healthy controls. The EC50 values of the untreated patients were significantly lower than in the controls. Lithium-treated patients had EC50 values significantly higher than controls. These results suggest that manic-depressive disorder is associated with an increased sensitivity of the inositol phospholipid second-messenger generating system, which is counteracted by lithium treatment. PMID- 8232837 TI - Calcium antagonists in manic-depressive illness. AB - Increased free intracellular calcium ion concentration ([Ca2+]i) has been found in lymphocytes and blood platelets of patients with bipolar affective disorders when they are acutely ill, but not after recovery. Because lithium alters intracellular calcium ion dynamics and lowers platelet [Ca2+]i in affectively ill patients but not controls, drugs whose primary action is to modulate [Ca2+]i in hyperactive cells have been used as antimanic agents. The best studied of these is verapamil, a calcium channel blocking agent (CCB) that appears most effective for lithium-responsive patients. Because they interact with different central CCB receptors, second-generation CCBs may have a different spectrum of action. CCBs are usually well tolerated and may be useful for a number of other psychiatric, neurological and medical conditions. PMID- 8232838 TI - Rapid cycling patients: conceptual and etiological factors. AB - The concept of rapid cycling is confusing in terms of its definition, clinical features, course of illness and the outcome. To complicate the matter further, rapid cycling has been reported to be drug resistant. Currently this condition has been reported to be secondary to lithium, tricyclic antidepressants and other agents such as cyproheptadine, a serotonin antagonist, piribedil, propranolol and levodopa. The increase in rapid cycling has coincided with the rapid increase in cocaine use in the society even though such an association cannot prove a causal relationship. Clinical or subclinical hypothyroidism as well as hyperthyroidism have been implicated in rapid cycling. In addition to the lack of specific knowledge on the etiology, a number of heterogeneous disorders has been grouped under this entity. It is useful to attempt a classification to ascertain whether clinically distinguishable subgroups have a common or different pathophysiology and to tailor the treatment that is most desirable for each subgroup. PMID- 8232839 TI - Clomipramine increases the responsiveness of raphe-cortical neurons in the rat. AB - The cerebral cortex of the rat possesses 5-HT1A receptor subtype; however, whether the activation of this receptor by dorsal raphe nucleus stimulation produces a similar action on neuronal cortical firing as antidepressants acting on serotonin (5-HT) function is unknown. Single-unit extracellular recordings were obtained from parietal cortex in rats in which dorsal raphe nucleus or cerebellar peduncle was stimulated. Results compared groups long-term treated with clomipramine (1.25 mg/kg i.p., twice a day, during 30 days) or saline (0.15 ml i.p., twice a day, during 30 days). Amplitude of traces, firing rate, and response to treatment with clomipramine proved to be different between cortical neurons responding to dorsal raphe nucleus or cerebellar peduncle stimulation. Clomipramine increased a long-lasting cortical suppression of firing elicited by dorsal raphe nucleus stimulation. Obtained data suggests that the responses of nearby cortical neurons containing different kinds of 5-HT receptors may relate to the involved pathways and that clomipramine seems to act on 5-HT1A receptor subtype. PMID- 8232840 TI - Probable cerebellar abnormality on N-isopropyl-(iodine-123)-p-iodoamphetamine single photon emission computed tomography scans in an epileptic patient receiving long-term high-dose phenytoin therapy. AB - Quantitative assessment of regional cerebral blood flow (rCBF) was performed in a 22-year-old man with intractable temporal lobe epilepsy on long-term high-dose phenytoin (PHT) therapy, using single photon emission computed tomography (SPECT) with N-isopropyl-(iodine-123)-p-iodoamphetamine. In all the SPECT scans repeated three times with changes in the PHT dose, absolute rCBF values were lower in most of the cerebral and cerebellar regions measured, as compared with 5 normal subjects matched for sex and age, and 22 normal subjects including the 5 men. Both the cerebellar to frontal rCBF ratio and cerebellar to cerebral rCBF ratio in the patient persistently showed low values, probably reflecting abnormal relative cerebellar hypoperfusion, whereas X-ray computed tomography and magnetic resonance brain imaging showed no abnormal findings. However, this probable cerebellar abnormality on SPECT was to some degree reversible with decreases in the PHT dose. These results suggest the utility of SPECT scans for early detection of cerebellar abnormalities known to be often present in epileptic patients, and imply a risk of long-term high-dose PHT therapy. PMID- 8232841 TI - Vascular dementia in the Lundby study. 2. An attempt to identify possible risk factors. AB - Vascular dementia represents a group of diseases of great medical and social impact. Its etiology is largely unknown but is most likely multifactorial. Factors related to the brain circulation are involved. The aim of the present study was to find out if personality and social background factors might have a clear precipitating or protective value. This study comprises a total population of 2,612 individuals from a geographically delimited area, Lundby, in southern Sweden. In 1957, Hagnell personally examined 99% of the population. A follow-up study of the same individuals, irrespective of domicile, was performed 15 years later. No social background factors proved to be relevant in this study. Personality factors, including psychosomatic reactions, did, however, demonstrate a highly predictive influence on the risk of developing vascular dementia. PMID- 8232842 TI - Investigation of the effects of coffee on alertness and performance during the day and night. AB - An experiment was carried out to examine the effects of coffee on performance and alertness in the day and at night. The results showed that caffeinated coffee had a beneficial effect on alertness and improved performance on a variety of tasks in both day and night sessions. The effects were often very large. For example, at night, consumption of caffeinated coffee produced comparable alertness ratings to the day-time ratings given when juice was drunk. In contrast to the effects of caffeinated coffee, the difference between the decaffeinated coffee and juice were small and variable. Overall, these results clearly demonstrate the beneficial effects of consuming caffeinated coffee, and show that this effect is comparable in the day and night. PMID- 8232843 TI - Quantitative electroencephalographic correlates of steroid administration in man. AB - Elevated levels of circulating corticosteroids are frequently associated with behavioral alterations in man, although the mechanisms by which corticosteroids may affect behavior are poorly understood. To evaluate possible effects of exogenous corticosteroids on brain electrophysiological functioning and the relationship of such effects to behavioral and biochemical changes, we administered prednisone (80 mg p.o. daily for 5 days) in a double-blind manner to 11 medically healthy volunteers. Quantitative electroencephalogram analysis was performed following 4 days of prednisone administration and during the preceding and ensuing placebo administration periods. Central theta wave brain electrical activity significantly increased following prednisone administration and returned to baseline following prednisone withdrawal. This effect was directly correlated with prednisone-induced increases in subjective sadness ratings and with decreases in self-rated energy and well-being. Prednisone-induced reductions in peak alpha wave activity were also directly correlated with increases in subjective sadness and Symptom Checklist-90 ratings and with decreases in self rated 'hypomanic' symptoms. Further, prednisone-induced increases in theta activity were significantly correlated with prednisone-induced decreases in CSF levels of somatostatin-like immunoreactivity, and prednisone-induced decreases in peak alpha activity were significantly correlated with decreases in CSF levels of norepinephrine and with relative increases (or lesser decreases) in CSF levels of beta-endorphin and beta-lipotropic hormone. This preliminary report of the concomitant development of prednisone-induced changes in brain electrical activity, neurochemistry and behavior highlights areas for future exploration in the study of corticosteroid effects on behavior in man. PMID- 8232844 TI - Segmentation of the spontaneous EEG in dementia of the Alzheimer type. AB - Studies on EEG data in dementia of the Alzheimer type have mainly investigated parameters in the frequency domain. However, the EEG also reflects the momentary functional state of the brain. The EEG can be partitioned into segments of spatially stationary map landscapes as characterized by the locations of potential maxima and minima, to study the time domain of the EEG. We calculated the segment structure of 19-channel EEG recordings (1-30 Hz) from 10 patients with dementia of the Alzheimer type and 10 sex-matched geriatric controls. The EEG maps corresponding to global field power peaks were used for analysis. The locations of the potential maximum and minimum of the maps were determined over the course of 20 s and used for segmentation. The segments of EEG activity were significantly fewer (p < 0.05) and the segments lasted longer (p < 0.05) in patients compared to controls. This effect could be observed over the whole period of recording and thus was not caused by only a few long segments. The data may be linked to morphological and biochemical findings of neuronal loss and loss of synapses in dementia of the Alzheimer type. PMID- 8232845 TI - Effects of a magnesium-deficient diet on sleep organization in rats. AB - The influence of magnesium, one of the most important cations in the vertebrate body, on the sleep-wakefulness cycle and ECoG patterns in chronically implanted rats recorded during the light period over a 6-hour period was investigated. Two groups of rats were studied. Group 1 (6 rats): after a control period of 2 weeks, the rats were maintained for 9 weeks on a Mg(2+)-deficient diet. Group 2 (5 rats): after a control period of 2 weeks, the rats were maintained for 7 weeks on a Mg(2+)-deficient diet followed by 4 weeks on a normal diet (recovery period: weeks 8-11). Mg(2+)-deficient diet for 9 weeks induced sleep and ECoG time dependent alterations. After 6-7 weeks on a Mg(2+)-deficient diet (n = 11) sleep analysis showed a significant increase of wakefulness (+50%) at the expense of slow wave sleep (-24%) but paradoxical sleep was not significantly modified. After 9 weeks of a Mg2+ deficient diet, sleep was disorganized: light sleep and polyspikes occurred indicating an increase in neuronal excitability. When Mg2+ was reintroduced in food and water, sleep organization and ECoG recordings were restored to their original patterns. Our findings which are in line with previous clinical and pharmacological observations provide conclusive arguments for the neuroprotective effect of magnesium ions in neurologic disorders and epileptiform activity. Mg2+ deficiency induces ECoG alterations in the rat which bear some similarities with those seen in neurogenic spasmophilic syndromes in man. PMID- 8232846 TI - Attentional dyslexia: a single case study. AB - A single case study of an attentional dyslexic is reported. The patient B.A.L. was able to read single letters and single words presented in isolation without difficulty. However, his reading of prose was very disrupted and his ability to read rows of letters and words was significantly impaired. A "flanking" procedure, in which there was a single target item, flanked by other stimuli, was used in a series of experiments to analyse his dyslexic impairment. First it was established that his attentional deficit was specific to reading in so far as the patient did not have comparable difficulties with pictorial material. His performance in the flanking experiments was consistently impaired; and furthermore was unaffected by speed of presentation, the number of flanking stimuli, and the spatial arrangement of the flanking stimuli. The important exception was when the flanking stimuli were of a different category. There was no decrement in performance when target letters were flanked by words or when words were flanked by letters. This study both corroborates and extends the original account of attentional dyslexia (Shallice and Warrnington, Neuropsychologia 15, 31-41, 1977) in terms of a damaged "filter" mechanism controlling the transition from a parallel to a serial stage of reading processing. The categorical effects reported here indicate that such a filter must be post lexical and suggest further that there are multiple filters in the reading system. PMID- 8232847 TI - Saccadic eye movements of dyslexic adult subjects. AB - The characteristics of visually guided saccadic eye movements were compared in 12 normal adult subjects and 12 test subjects of normal intelligence, but with problems in reading and writing. All subjects were examined psychometrically for different cognitive abilities, and for their reading and writing capabilities. The anamnestic reports about their reading and writing problems earlier in their lives were analysed. Based on scores of the writing and reading tests, the test group was subdivided into two subgroups: A and B (group A reached medium, group B very low scores in both tests). Five different non-cognitive eye movement tasks were applied: two single tasks (gap and overlap) requiring single saccades from a fixation point to a peripherally appearing target and three sequential tasks (overlap, synchronous, and simultaneous) requiring sequences of saccades to four equally spaced targets presented sequentially to the right side from an initial fixation point. Many parameters of the subjects' eye movement performance were determined and their mean values were calculated for each subject. The Student t test revealed that the eye movement data of the two test groups deviated differently from the data of the control group. Group B had the largest deviation of the eye movement parameter from the control group. The differentiating parameters were the consistency of target acquisition, the saccadic reaction time, and the number of anticipatory responses in the single target tasks. In the sequential tasks these differences were in the amplitude, the number of saccades, and the fixation durations. The incidence of regressive saccades did not discriminate between test subjects and controls. The possible underlying deficits in the attentional control over the saccadic system and their implications for reading are discussed. PMID- 8232848 TI - Investigations of the functional anatomy of attention using the Stroop test. AB - In two separate experiments positron emission tomography (PET) was used to measure changes in regional cerebral blood flow while normal subjects performed the Stroop colour word interference test, a test of selective attention. In the first experiment performance of the Stroop task was associated with activation of right orbito-frontal and bilateral parietal structures, an unexpected result in view of previously reported findings. In addition, there were highly significant time related focal changes in rCBF. A second experiment was therefore carried out which altered the experimental parameters to replicate an earlier study. In this second experiment focal activation of the right anterior cingulate and right frontal polar cortex occurred during the Stroop task. As in the first experiment significant time effects were again apparent. To determine the functionally related brain systems during the performance of the Stroop task a correlation analysis was carried out in relation to blood flow changes induced by experimental manipulation in the right anterior cingulate. This analysis indicated the engagement of a widespread network of anterior brain regions and reciprocal inhibition of posterior brain regions during the performance of the task. The results provide evidence for the involvement of anterior right hemisphere and medial frontal structures in attentional tasks but also indicate that time effects can confound task specific activations. Furthermore subtle experimental treatment parameters, such as stimulus presentation rate, influence the degree and distribution of observed activations. PMID- 8232849 TI - Visual and auditory cortical lesions following acquisition of an intensity discrimination in rats fail to disrupt cross-modal transfer. AB - The effect of visual or auditory decortication on cross-modal transfer of an intensity discrimination was examined in rats. Twenty animals were first trained under either visual-auditory (V-A) or auditory-visual (A-V) cross-modal transfer (CMT) in a shuttlebox using a shock avoidance pardigm. Prior to the second training session, five of the A-V animals received auditory ablations and five V A animals received visual ablations. The other 10 animals served as controls and received sham operations. The results reveal that CMT occurred in both experimental groups following cortical ablations. It is possible that information regarding stimulus intensity was transferred from a cortical region used during the original training session to the cortex used in the second or retraining session, prior to surgery. Alternatively, it may be that some subcortical structure (e.g. the amygdala, superior colliculus, or reticular formation) may be involved in CMT of intensity. PMID- 8232850 TI - Short-term memory and temporal ordering in early Parkinson's disease: effects of disease chronicity and medication. AB - Studies of Parkinson's disease (PD) have shown impaired temporal ordering but interpretation may be confounded by task requirements and the effects of medication. We examined item recognition and recency discrimination in PD in relation to treatment and performance on other tests. Patients showed increased response latency and impaired recency discrimination only at short retention intervals. The deficits were greater in chronically medicated patients but treatment with levodopa, bromocriptine or anticholinergic drugs did not affect performance of newly diagnosed cases. The short-term memory deficits correlated with scores on tests of working memory, attention and executive function. These results do not indicate a generalised temporal ordering deficit in PD but suggest that much of the cognitive impairment in the disorder arises from attentional deficits affecting short-term and working memory. PMID- 8232851 TI - Algorithmic and heuristic strategies in comprehension of complement clauses by patients with Parkinson's disease. AB - Language ability in patients with idiopathic Parkinson's disease (PD) and normal control subjects (NC) matched on age, sex, education and socioeconomic status (SES) was investigated. The two groups of subjects were tested on eight sentence types in Greek in the form of main and complement clause with eight matrix verbs. These matrix verbs were ask (ask information), promise (commissive meaning), tell1 (order, command) and tell2 (give information) in sentences with no semantic constraints, and confess, sell, trust and scold in sentences with semantic constraints (implicit causality). The results show that language ability, despite relative preservation is significantly impaired in PD patients as compared to that of NC. More specifically, syntax with semantic constraints was the most effective independent variable to classify PD patients and NC subjects into two distinct groups according to a Logistic Regression Analysis. To restrict the algorithmic process in sentence comprehension, PD patients seem to make use of the minimal distance principle (MDP) and the "experiencer constraint" heuristic strategies. Possible similarities in language behavior between PD patients and aphasics, in general, are suggested. PMID- 8232852 TI - Processing of tactile stimuli and implications for the reading disabled. AB - A finger localization task was used to investigate interhemispheric transfer of tactile information and the processing of sequential tactile information in 7- and 9-year-old "average" readers and 9-year-old reading disabled boys. The performance of the reading disabled group on interhemispheric transfer and sequential processing was comparable to or worse than the 7-year-old average readers. Significant correlations linked the reading disabled boys' performance on the finger localisation task with their reading skills, performance on perceptual/motor tasks and knowledge of order in the time domain. The results are interpreted in terms of delayed myelination of interhemispheric and intrahemispheric pathways in the reading disabled. PMID- 8232853 TI - Olfactory identification deficits in Down's syndrome and idiopathic mental retardation. AB - We investigated olfactory identification in children and adults with Down's syndrome (DS) and idiopathic mental retardation (IMR) and in age-matched normal controls (NC). Identification was assessed with a four alternative-forced-choice task modified from the University of Pennsylvania Smell Identification Test (M UPSIT) and a yes/no task yielding measures of discrimination and response bias for the same stimulus material. Control tactile identification tasks were also administered. Results were that odor identification performance on both tasks was specifically impaired in DS compared to IMR and NC. Accuracy of identification on the M-UPSIT correlated inversely with age in DS only. When uncertain, DS and IMR subjects guessed "yes" more often than "no" on the Yes/No task (liberal decision bias) and guessed the last response alternative on the M-UPSIT (recent position bias), whereas the normal subjects had neutral decision bias on the Yes/No task and matched the objective position presentation probabilities on the M-UPSIT. Decision bias correlated with accuracy of identification in both tasks for the DS subjects only. PMID- 8232854 TI - Ocular dysfunction during the amytal suppression test. AB - We assessed the following parameters of visual function in 16 patients during amytal suppression test (AST): pupillary light response, consensual reflex, pupillary symmetry, monocular reading and monocular visual recognition. Significant visual impairment in the eye ipsilateral to injection (EII) occurred in 7 of 16 patients ranging from blurring to complete, transient loss of vision. We conclude that visual dysfunction of the EII during the AST is common and represents an important component of the test. Intracarotid amytal reaches the EII via ophthalmic artery and probably impairs vision in the EII due to cycloplegia, iridoplegia and retinal suppression. PMID- 8232855 TI - Gaze motor asymmetries in the perception of faces during a memory task. AB - In 33 male and female adult volunteers, eye position recordings were performed by means of an infrared reflection technique. Slides of randomly shuffled black-and white photographs (7.5 x 10 degrees) of faces and vases were projected for 6 or 20 sec respectively in a visual memory task. In each series, 10 slides of art nouveau vases and of the "inner part" of masked Caucasian faces were used. During recording the head was fixed by a bite-board. (a) For faces the preferred targets of the centre of gaze were the eyes, the mouth and nose region, for vases the contours and some prominent ornaments. (b) Left-right asymmetries in the gaze movement sampling strategy appeared with faces, but not with vases. In faces, the overall time that the centre of gaze remained in the left half of the field of gaze was significantly longer than in the right half. (c) When, however, the amplitude of the gaze excursions into the left and right halves of the inspected items was taken as a measure and normalized, a preference for the right gaze field was observed. (d) The relative left-right bias during face inspection was stronger with the 6 sec than with the 20 sec inspection period and significantly stronger in female than in male subjects for the 6 sec tasks. (e) Left/right inversion of the face stimuli did not abolish the side bias. Thus the asymmetric sampling strategy when faces were inspected as compared to vases was due to "internal" factors on the part of the subjects. It is hypothesized that a left right asymmetry in hemispheric visual data processing for face stimuli was the cause of a left-right asymmetry in gaze motor strategies when faces were inspected. PMID- 8232856 TI - The functional role of an increase in cell excitability and synaptic efficiency in the new cortex during learning. AB - Data were obtained in experiments on nonimmobilized and nonanesthetized rabbits, during the development of an analog of a CR, with recording of the response of the pyramidal tract, suggesting temporal specificity in the manifestations of membrane and synaptic plasticity, the participation of these mechanisms in both representations of the combined stimuli, and primarily unidirectional changes in the degree of their participation in these points of the cortex. It is concluded that temporary membrane plasticity creates conditions through the mechanisms of synchronization and summation for the passage of excitation from the sensory link to the motor output of the new connection. The gradual reorganization of excitatory and inhibitory connections to the output elements of the conditioned reflex act, determined by the mechanisms of synaptic plasticity, determine and strengthen the specialized character of the developed reaction. PMID- 8232857 TI - The EEG analysis of the interrelationships of structures of the thalamofrontal system during the recovery of conditioned reflex behavior of amygdalectomized rats. AB - The influence of a unilateral lesion of the basolateral amygdala (BLA) on instrumental reflexes and the EEG activity of the orbitofrontal cortex (OFC) and the dorsomedial nucleus of the thalamus (DMN) was investigated in experiments on rats. A decrease in the number of correct reactions and an increase in their latency over the course of the first two postoperative weeks was accompanied by an increase in the amplitude of the delta and theta rhythms in the EEG of the OFC and DMN, whereas in the following two weeks this change appeared only in the EEG of the OFC. The changes in the EEG reflecting new functional relationships between the structures of the injured system were more prolonged (more than a month) than the period of the recovery of the parameters of the reflexes. It is hypothesized that an important role in the foundation of the disturbance and recovery of the instrumental reflex behavior following destruction of the BLA belongs to the functional reorganization of connections between the limbic and thalamocortical structures of the brain. PMID- 8232858 TI - The dependance of neuronal reactions of the sensorimotor cortex to a simultaneous complex stimulus upon the level of differentiation of its components. AB - The change in the neuronal activity of the sensorimotor area of the cerebral cortex of the cat was investigated in awake animals as a function of the level of differentiation of the components of a simultaneous heteromodal complex stimulus. Two groups of neurons in the sensorimotor cortex were distinguished on the basis of the character of this relationship and a number of other parameters. It was shown that the parameters of the reactions of all neurons recorded to the positive conditional stimulus following the consolidation of the conditioned motoric reaction are established first. Such parameters of the responses as degree of manifestation, intensity, duration, and the length of the latent period changed in the process of development. The reactions of neurons of both groups to inhibitory signals were stabilized only after the consolidation of the differentiation skill. In the process only the pattern of the discharge changed in the neurons of the first group, while in the neurons of the second group, the degree of manifestation of the response, its sign, duration, and length of the latent period could vary. Fluctuations in the level of differentiation following the development of the inhibitory conditioned reactions had an effect only on the responses of the neurons of the second group to the components of the complex. PMID- 8232859 TI - The morphofunctional characteristics of neurons of the sensorimotor cortex of aged rabbits during the trace assimilation of rhythm. AB - The formation of trace rhythm assimilation (an analog of the conditioned reflex to time) by neurons of the sensorimotor cortex in response to prolonged (10-20 min) electrodermal stimulation of an extremity at a frequency of 0.5-1 or 2 Hz was investigated in awake adult (5-7 months), old (54-65 months), and very old (66-85 months) rabbits. The data of the spectral analysis of the impulse activity of 460 neurons showed aged-related differences in the formation of trace rhythm assimilation by neurons: it takes place after one to two series (1st to 2nd days of the experiment) in young animals; after two to four series of periodic stimulation (2nd to 3rd days of the experiment) in the old animals; in very old animals rhythmic stimulation essentially did not result in rhythm assimilation by cortical neurons. Quantitative and qualitative morphological changes in nerve and glial cells in the sensorimotor cortex of old rabbits as compared with young rabbits were identified. It is hypothesized that deviations in the course of mnestic processes observed in animals during aging may be associated with destructive phenomena in the neocortex during normal aging. PMID- 8232860 TI - The reactivity and interaction of neurons of the cerebral cortex of rats in the presence of the microiontophoretic action of acetylcholine in a model negative learning situation. AB - The activity of 46 pairs of neurons in the sensorimotor cortex of the brain of rats during multiple (up to 60) brief microiontophoretic applications of acetylcholine (ACh) was investigated by means of the method of multicellular recording, with subsequent division into separate impulse streams. It was demonstrated that neurons with high-amplitude spikes have a lower average frequency of discharges in comparison with low-amplitude [neurons]. It was established that cells located adjacent to one another exhibit responses in 63% of cases which are similar in their component makeup; the majority of these responses are inhibitory-excitatory and excitatory. Analysis of the dynamics of the structure of the cross-interval histograms plotted for pairs of cells on the basis of the data of their initial activity, as well as activity in the baseline, and upon termination of the action of ACh, attest to the relative invariability of the excitatory connections between the cells and a possibility of some increase in the efficiency of the inhibitory connections under the conditions of the manifestation of the dynamics of responses to ACh of habituation type by one of the cells. PMID- 8232861 TI - The influence of exogenous gangliosides on the dynamics of the development of prolonged posttetanic potentiation. AB - The influence of various gangliosides on the dynamics of the development of prolonged posttetanic potentiation (PPTP) in the layer of pyramidal cells of field CA3 during the stimulation of mossy fibers was investigated in rat hippocampus section. It was demonstrated that the GM1 and GD1b gangliosides stimulate the induction and maintenance of PPTP to varying degrees and that GD1a and GT1b inhibit the phase of maintenance as compared with the active control. A conclusion was reached regarding the participation of gangliosides in the mechanisms of synaptic plasticity. PMID- 8232862 TI - The reinforcing function of the emotions. AB - The present study is devoted to a brief exposition of the results of investigations of the central mechanisms of the reinforcing properties of emotions that were carried out in the I.P. Pavlov Physiology Department of the Institute of Experimental Medicine in recent years, and to an attempt to outline the prospects for further developments in this sphere of brain physiology which is vital for conditioned reflex theory. PMID- 8232863 TI - The amygdaloid complex in the sex differentiation of the brain. PMID- 8232864 TI - The influence of immune suppression on the take rate and development of the human neocortex transplanted to the spinal cord of the adult rat. PMID- 8232865 TI - The influence of injections of 6-hydroxydopamine into the nucleus accumbens and the substantia nigra on the behavior of rats. AB - Bilateral injections of 6-hydroxydopamine into the nucleus accumbens of rats with destroyed dopaminergic neurons of the substantia nigra leads to an increase in the horizontal motoric activity in the open field and accelerates the appearance of conditioned reflex responses in the course of the postoperative restoration of the skill as compared with these indices in rats in which the neurotoxin was injected only into the substantia nigra. Thus, destructions of the dopaminergic system of the nucleus accumbens can compensate to a certain degree for the consequences of nigrostriatal pathology. PMID- 8232866 TI - An investigation of the effect of amtizol on the plastic properties of the membrane of the Retzius neuron of the leech. AB - The effect of amtizol, a blocker of the inactivation of calcium-dependent potassium channels of the outward current, on the spontaneous and evoked impulse activity of these cells was investigated in experiments on Retzius neurons of the leech. It was demonstrated that the extracellular application of amtizol induces a decrease in the amplitude and an increase in the duration of evoked AP, while not exerting a perceptible influence on the form of the spontaneous AP. In addition, disruption of the development of the process of habituation that develops in the norm during high-frequency synaptic activation of the neuron is observed in a solution containing amtizol. PMID- 8232867 TI - The influence of the delta-sleep-inducing peptide on convulsive activity. AB - Data are presented in this paper on the influence of the delta-sleep-inducing peptide (DSIP) on various forms of convulsive activity. The capacity of this peptide to suppress convulsive activity in foci created in the cerebral cortex by the application of strychnine has been demonstrated in experiments on cats. It has been established in experiments on rats that DSIP determines the later development of the convulsive kindling syndrome, and prevents the development of convulsions in mice induced by bicuculline, picrotoxin, and corazol, but is devoid of such action in relation to thiosemicarbazide and strychnine. It was demonstrated that the anticonvulsant action of DSIP is associated with its influence on the reticular portion of the substantia nigra. The lowest level of this peptide itself has been discovered, in the reticular portion of the substantia nigra at the late stages of pharmacological kindling. It is inferred that DSIP may represent one of the factors of the endogenous control of the excitability of the brain. PMID- 8232868 TI - Electrophysiological and cognitive effects of lumbar myelography with iopamidol: comparison with diagnostic lumbar puncture. AB - To assess the influence of contrast medium on cortical function, we studied 20 patients undergoing lumbar myelography with iopamidol and 10 patients undergoing diagnostic lumbar puncture (controls). The examinations performed before and 6 and 24 h after myelography (or lumbar puncture) included a neuropsychological battery and an electrophysiological evaluation. In the patients cranial CT was performed thrice to assess passage of contrast medium from the cerebrospinal fluid into the brain. Neither patients nor controls had significantly different scores on neuropsychological testing after the diagnostic examinations. A transient slowing of basal EEG activity could be detected in 2 patients and 3 controls 6 h after the lumbar puncture. In 3 patients CT showed a transient increase in density of the brain. None of the parameters studied was significantly affected by myelography with iopamidol. CT findings support the hypothesis of early clearance of iopamidol from brain tissue, explaining its low neurotoxicity. PMID- 8232869 TI - The contribution of needle size and other factors to headache following myelography. AB - The author performed a randomized prospective study of the effects of needle size and of age, sex, myelogram type and operators on the incidence of headache following myelography. In 113 patients randomized to 22 G and 25 G needle groups, data were obtained by questionnaire on the frequency of headache up to five days following the procedure. The influence of needle size was statistically insignificant (63%, 48%, p > 0.05). Age (p = 0.0002), sex (p = 0.01), myelogram type (p = 0.002) and myelographer (p = 0.03) significantly affected the frequency of headache. The size of the dural perforation appears to be less important than factors relating to age, sex, myelogram type and individual myelographers in the development of headache after myelography. PMID- 8232870 TI - Gadodiamide injection as a contrast medium for MRI of the central nervous system: a comparison with gadolinium-DOTA. AB - The tolerance and diagnostic efficacy of a new nonionic, intravenous MRI contrast medium, gadodiamide injection (Omniscan) are reported and compared with those of gadolinium-DOTA after completion of a double-blind, randomized, parallel study in 60 patients using a dose of 0.1 mmol/kg. Two patients were excluded from the statistical analysis after trial entrance since no contrast medium was injected. The indications for the MRI examination were known or suspected CNS lesions. Vital signs, chemical and haematological parameters were monitored. No significant abnormalities were observed, confirming the excellent tolerance of both contrast media. The usefulness of gadolinium-containing contrast media was again documented, since in 8.6% (5/58) of the patients the lesion(s) could be seen only after injection. Contrast enhancement was seen in 87.8% (43/49) of the patients with abnormal findings, and provided additional diagnostic information in 24.5% (12/49). Lack of contrast enhancement in 12.2% (6/49) of patients with abnormal findings was a significant negative finding. PMID- 8232871 TI - MRI of an abscess of the cervical spinal cord in a case of Listeria meningoencephalomyelitis. AB - A case of Listeria meningoencephalomyelitis is described and the MRI appearances of an abscess of the cervical spinal cord are presented. The MRI appearances did not allow a definitive diagnosis before death. PMID- 8232872 TI - Epidural spinal abscess containing gas: MRI demonstration. PMID- 8232873 TI - Aortic pathology revealed by MRI in patients with clinical suspicion of spinal disease. AB - In five patients with clinical suspicion of spinal disease, MRI of the spine revealed unexpected aortic pathology explaining the symptoms. No significant intraspinal pathology was found on MRI. However, in one patient with clinical suspicion of spinal stenosis, an aortic occlusion was detected on MR images of the spine. The lower extremity ischaemia, caused by the occlusion, was responsible for the symptoms. In another patient a paravertebral haematoma from a ruptured aortic aneurysm resulted in spinal nerve compression, thought before MRI to be caused by a spinal tumour. In three patients aortic aneurysm or dissection resulted in spinal cord ischaemia with symptoms mimicking those of compressive spinal disease. Thus, if MRI of the spine does not provide an explanation for the patient's symptoms, examination of the aorta is recommended. PMID- 8232874 TI - Gadolinium enhancement of vertebral endplates following lumbar disc surgery. AB - Gadolinium enhancement may be normal in the vertebral endplates adjacent to previously operated intervertebral discs. To determine the frequency of this finding and to differentiate this normal healing process from early lesions due to focal infection, we studied 135 patients who had undergone surgery for herniated lumbar disc, and compared them with 249 unoperated patients with radicular symptoms and 15 with known spondylodiscitis. Hypointense foci which enhanced with gadolinium were identified in the endplates of 25 (18.5%) of the operated patients, 9 of whom required a second operation for recurrent disc herniation, at which time the absence of infection was confirmed. Gadolinium enhancement within the endplates adjacent to the operated disc occurs during normal healing after surgery. Care should be taken before invoking a diagnosis of focal infection or secondary spondylodiscitis. PMID- 8232875 TI - Contrast-enhanced MRI of healed pathologic vertebral compression fracture mimicking active disease in a patient treated for lymphoma. AB - Despite the valuable contribution of MRI, the distinction between benign and malignant compression fractures of the spine is sometimes difficult, and cannot with certainty be based on T1- and T2-weighted signal criteria alone, or on the configuration of the compression fracture. This case report demonstrates that in certain instances neither can gadolinium enhancement specifically make this determination in cases of treated malignancy of the spinal column. Because treated neoplasia having benign histologic characteristics may continue to enhance with IV gadolinium, biopsy evaluation must be undertaken in this setting in order to prove the diagnosis. PMID- 8232876 TI - Granulocytic sarcoma (chloroma) causing spinal cord compression. AB - Granulocytic sarcoma (chloroma) is a rare solid tumor of myelogenous stem cells, usually appearing in patients with acute myelogenous leukemia and less commonly in patients with chronic myelogenous leukemia or myeloproliferative disorders. We present a spinal epidural granulocytic sarcoma causing thoracic spinal cord compression in a patient with chronic anemia secondary to myelofibrosis. PMID- 8232877 TI - Correlation of MRI and clinical features in meningeal carcinomatosis. AB - Ten patients with meningeal carcinomatosis associated with nonhaematological neoplasms were examined: six with breast, two with gastrointestinal and one with lung cancer, plus one with a tumour of unknown origin. Cytology was positive in all but one. The patients were classified into four groups according to the gadolinium-enhanced MRI (Gd-MRI) appearances: group 1 had pure leptomeningeal carcinomatosis, group 2 dural carcinomatosis, group 3 spinal leptomeningeal carcinomatosis, and group 4 had normal Gd-MRI except for hydrocephalus. In group 1, Gd-MRI showed diffuse enhancement of the subarachnoid space, including the cisterns around the midbrain, the sylvian fissures, or cerebellar and cerebral sulci. In group 2, Gd-MRI showed diffuse, thick, partially nodular enhancement of the dura mater. No leptomeningeal or subependymal enhancement was evident. In group 3, nodular masses were seen only in the spinal canal. In group 4, no definite evidence of meningeal carcinomatosis was demonstrated on contrast enhanced CT (CE-CT) or Gd-MRI. The median survival time was 2.0 months in group 1, 1.0 month in group 3, and 4.5 months in group 4, but the two patients in group 2 were alive 10 and 15 months after a definite diagnosis of meningeal carcinomatosis was made. In all patients examined by both CE-CT and Gd-MRI, the latter was superior for identification of meningeal carcinomatosis. Hydrocephalus in an important indirect sign of leptomeningeal carcinomatosis, but was not seen in patients with dural carcinomatosis despite the presence of increased intracranial pressure. PMID- 8232878 TI - Myelography, CT and MRI in leukaemic infiltration of the lumbar theca. AB - A 25-year-old woman with acute lymphoblastic leukaemia, while in remission, developed paraparesis, with faecal and urinary incontinence. CT demonstrated increased density of the lumbar theca and enlargement of the nerve roots. Myelography showed complete obstruction below the L3 level. MRI showed increased signal intensity in the lumbar sac on T1 weighting, and the cauda equina enhanced with gadolinium-DTPA. Lymphoblasts were seen in the lumbar spinal fluid. After chemotherapy, these abnormalities resolved, as did the paraparesis and incontinence. PMID- 8232879 TI - Large thoracocervical intraspinal neuroma: unusual clinical and neuroradiological findings. AB - The clinical and radiological findings of a giant cystic intraspinal cervical tumour are presented. Symptoms and signs suggesting a polyneuropathy were misleading. On myelography the tumour appeared as a solid mass, but its size was distinctly underestimated. MRI demonstrated the true extent (from C4 to T4) and the cystic structure of the tumour. The histo-pathological diagnosis was fibrillary neuroma (WHO grade I). PMID- 8232880 TI - CT in autosomal dominant and idiopathic cerebellar ataxia. AB - Signs of atrophy on cranial CT were investigated in 35 patients diagnosed as suffering from autosomal dominant (n = 21) or idiopathic (n = 14) cerebellar ataxia. Thirteen patients with a pure cerebellar syndrome were examined after at least 4 years of disease (mean duration 10.5 years) and were classified as cerebellar atrophy (CA). Twenty-two patients with additional non-cerebellar signs were classified as olivo-ponto-cerebellar atrophy (OPCA). Four (30%) of the patients with CA had atrophy of the brain stem in addition. Of the 22 patients with OPCA, 9 (40%) had atrophy of the cerebellum only. In patients with CA or OPCA correlation of clinical signs with severity of atrophy on CT was poor. Atrophy on CT often fails to differentiate autosomal dominant or idiopathic cerebellar ataxias in CA or OPCA: patients with CA can also have atrophy of the brain stem and patients with OPCA do not necessarily show brain stem atrophy. PMID- 8232881 TI - SPECT, CT and MRI in a Turkish family with Huntington's disease. AB - A Turkish family with Huntington's disease documented on CT, MRI and SPECT is reported. Whereas in clinically definite cases CT and MRI are of limited value and SPECT does not add anything of value, in one asymptomatic subject SPECT showed moderate caudate nucleus hypoperfusion, underlining the hypothesis that SPECT may have a role in predicting Huntington's disease. PMID- 8232882 TI - Lyme disease presenting as a stroke in the vertebrobasilar territory: MRI. AB - A 28-year-old female farmer, without vascular risk factors, developed a limited infarct of the pons, associated with a lymphocytic cerebrospinal fluid (CSF) pleocytosis. Titres of specific antibodies against Borrelia burgdorferi were high in serum and CSF. MRI confirmed an infarct in the territory of the medial pontine arteries, but angiography showed no evidence of cerebral angiopathy. Antibiotic therapy rapidly led to a return to normal of CSF cytology and serology. We suggest that Lyme disease is a possible cause of cerebral ischaemia. PMID- 8232883 TI - MRI of intracranial meningiomas: correlations with histology and physical consistency. AB - Magnetic resonance imaging (MRI) appearances of 43 intracranial meningiomas were reviewed and correlated with their histology and consistency at surgery and on pathological examination. There was a statistically significant prevalence of hard tumours within the fibroblastic subtypes. No statistically significant correlation between signal intensity and histology or consistency was found. A meningioma hyperintense on T2-weighted images is, however, unlikely to be fibroblastic or hard. PMID- 8232884 TI - Stapedial artery supplying sphenoid wing meningioma: case report. AB - A 46-year-old woman presented complaining of resting tremor of the right arm and attacks of inappropriate crying and laughing. Computed tomography and magnetic resonance imaging of the head revealed a large left sphenoid wing meningioma. Preoperative cerebral angiography demonstrated a persistent stapedial artery, terminating as a middle meningeal artery which was the main blood supply to the tumour. PMID- 8232885 TI - Pineal meningioma: MRI. AB - Pineal meningiomas are very rare. We report a pineal meningioma examined by MRI. PMID- 8232886 TI - Problems with the "dural tail" sign. AB - Prominent enhancement of the dura mater, the "dural tail" adjacent to a peripherally located mass on gadolinium-enhanced MRI has been described as being characteristic of meningiomas. We present a cerebral glioma showing the classical "dural tail". PMID- 8232887 TI - MRI of cranial chordomas: the value of gadolinium. AB - Two intracranial chordomas are presented: a typical chordoma and a chondroid tumour. MRI with gadolinium showed different enhancement patterns, which may be related to pathological findings. The value of differentiating the two types of chordoma lies in their very different prognosis. PMID- 8232888 TI - MRI appearances of an ectopic pituitary adenoma: case report and review of the literature. AB - MR images of an ectopic pituitary adenoma have been published in only a single case, a suprasellar mass. We present a patient with Cushing's disease in whom MRI revealed a pituitary adenoma within the sphenoid sinus. Radiologic characteristics of ectopic pituitary adenomas are reviewed, with emphasis on MRI, which demonstrates a soft tissue mass, isointense with gray matter on T1-weighted images, which enhances in a heterogeneous manner. PMID- 8232889 TI - MRI visualization of complete bilateral optic nerve involvement by pituitary adenoma: a case report. AB - With large pituitary adenomas, the optic nerves and chiasm usually lie on the tumour capsule and are displaced superiorly. We report a large invasive pituitary adenoma, with complete involvement of both optic nerves. Review of the preoperative MR images demonstrated the optic nerves, with signal intensity close to that of cerebral white matter, and different from the flow void of the basal cerebral arteries. Correlation of this observation with intraoperative findings is discussed. PMID- 8232890 TI - MRI demonstration of cervical meningocele. AB - The MRI appearances of a cervical meningocele are reported. The operation demonstrated the accuracy of the preoperative MRI diagnosis. PMID- 8232891 TI - Intracranial calcification in siblings with nephrogenic diabetes insipidus: CT and MRI. AB - Computed tomography and magnetic resonance imaging (MRI) were used to examine three male siblings with nephrogenic diabetes insipidus (NDI). The two elder brothers had varying degrees of unusual intracranial calcification; the eldest also showed involvement of the cerebral white matter on MRI. The severity of intracranial calcification was related to the time before initiation of treatment and inversely to mental ability. Brain damage and mental retardation in NDI may be caused by a delay in initiating treatment; early detection and treatment are important to prevent brain damage. PMID- 8232892 TI - Cranial MRI and MR angiography in Menkes' syndrome. AB - We report two boys with Menkes' syndrome who underwent cranial MRI and MR angiography (MRA). In both, CT and MRI revealed progressive cerebral atrophy with a subdural haematoma or effusion. Delayed myelination or dysmyelination of the white matter was suggested. Tortuosity of the cervical and intracranial vessels was well demonstrated by MRA, obviating more invasive conventional angiography should it be thought necessary to demonstrate the characteristic systemic vascular changes of this syndrome. PMID- 8232893 TI - Cerebral MRI in 3-hydroxy-3-methylglutaryl-coenzyme A lyase deficiency: case report. PMID- 8232894 TI - Multiple arteriovenous fistulae in the Klippel-Trenaunay-Weber syndrome (KTW) PMID- 8232895 TI - Distribution and axonal relations of macrophages in a neuroma. AB - After axotomy in the peripheral nervous system, most axons regrow and re establish contact with their targets. Depending on the type of lesion, a varying number of nerve fibers fail to regenerate and terminate far from the target, forming a neuroma. Sensory axons trapped in a neuroma show abnormal sensitivity to various stimuli, and often fire spontaneously. In this study we have examined the distribution and axonal relations of macrophages in rat sciatic neuromas three days to one year after cutting and ligating the nerve. ED1-immunoreactive macrophages migrated into the neuroma in large numbers within the two first weeks after the injury. Most cells were at that time located 0.5-1 mm proximal to the ligature. From three weeks on, a majority of the ED1-immunoreactive cells contained numerous large vacuoles filled with myelin fragments. At sites of focal demyelination, macrophages often had direct contact with axonal membranes. At later survival stages (three months to one year) ED1-immunoreactive cells were seen not only in the area just proximal to the ligature, but also several millimeters proximal to this. Macrophages persisted in considerable numbers in the neuroma for at least one year. These data suggest that neuroma macrophages may participate in the genesis of electrophysiological abnormalities thought to underly chronic pain after neuroma formation, possibly by creating demyelinated axonal regions susceptible to external stimuli from e.g. neighboring nerve fibers, by releasing substances which influence regeneration and remodelling of axonal growth cones, or by direct actions on the denuded axonal membranes. PMID- 8232896 TI - Further substantiation of a significant role for the sympathetic nervous system in inflammation. AB - This study provides significant new evidence substantiating a role of the postganglionic sympathetic neuron in plasma extravasation in the knee-joint of the rat. Increased plasma extravasation produced by the potent inflammatory mediator bradykinin was mimicked by 6-hydroxydopamine, a selective stimulator of sympathetic fibers. Various treatments (chemical sympathectomy, co-perfusion with the local anesthetic lidocaine, or co-perfusion with depolarizing concentrations of potassium) similarly modulated plasma extravasation induced by both bradykinin and 6-hydroxydopamine, but not that produced by platelet activating factor. We also showed that bradykinin is able to release norepinephrine in the knee-joint, indicating action on the sympathetic postganglionic neuron. In summary, these experiments provide substantial additional evidence supporting a significant contribution of the sympathetic post-ganglionic neuron terminal to inflammatory plasma extravasation. PMID- 8232897 TI - Time-course analysis of changes in calcitonin gene-related peptide-and methionine enkephalin-immunoreactivity in the female rat preoptic area after estrogen treatment. AB - The time-course effects of one month of estrogen upon calcitonin gene-related peptide - and methionine-enkephalin-immunoreactivity in the periventricular preoptic nucleus and medial preoptic nucleus were semi-quantitatively investigated with a computer-based image analysis system. Female Wistar rats were ovariectomized and implanted subcutaneously with a 10-mm-long silastic capsule containing estradiol-17 beta, or with a blank capsule, as a control. Estradiol-17 beta-treated rats were killed at days 1, 4, 7, 10, 14 and 28 after the implantation of estradiol-17 beta. To investigate the details of changes in calcitonin gene-related peptide- and methionine-enkephalin-immunoreactive fibers in the periventricular preoptic nucleus and medial preoptic nucleus, a grid, made up of 8 x 16 squares (one square corresponding to 50 x 50 microns in the true section), was set on the wall of the third ventricle, and immunoreactivity within each square was measured with an image analyser. In the control rats, calcitonin gene-related peptide- and methionine-enkephalin-immunoreactive fibers were distributed in the periventricular preoptic nucleus and medial preoptic nucleus. In the estradiol-17 beta-treated rats, calcitonin gene-related peptide immunoreactive fibers increased prominently at day 1, day 7 and day 10 in the periventricular preoptic nucleus, whereas methionine-enkephalin-immunoreactive fibers increased at day 1, day 14 and day 28 in the periventricular preoptic nucleus and medial preoptic nucleus. These findings suggest that the mechanism underlying the increases in these calcitonin gene-related peptide- and methionine enkephalin-immunoreactive fibers after estrogen treatment might be different. PMID- 8232898 TI - The distribution of histaminergic axons in the superior colliculus of the cat. AB - The histaminergic projection from the hypothalamus to the superior colliculus was examined immunohistochemically in the cat brain using an antibody to histamine. The source of histaminergic fibers in the brain is a group of neurons in the posterior hypothalamus, located primarily in ventrolateral and periventricular regions and collectively referred to as the tuberomammillary nucleus. All laminae of the superior colliculus--including the superficial, intermediate, and deep layers, as well as the central gray--were blanketed with histamine-immunoreactive axonal fibers. Overall, labeling in the superior colliculus was moderately dense compared to other locations in the cat brain, with some variation in fiber density. Individual labeled fibers resembled histaminergic fibers described previously in the brain. Labeled axonal fibers showed infrequent branching and were beaded with numerous en passant varicosities that were typically 1 micron or smaller, but as large as 2.5 micron in diameter. Varicosity size differed significantly at different depths in the colliculus. The histaminergic projection appears to be separate from a previously reported, apparently non-histaminergic projection from neurons in the dorsal hypothalamic area to discrete regions of intermediate and deep colliculus. These results indicate that the histaminergic projection from the tuberomammillary nucleus of the hypothalamus projects extensively throughout the superior colliculus. Histamine, which is believed to act as a neuromodulator in the brain, is in a position to influence sensory and motor-related processes in every layer of the cat superior colliculus. PMID- 8232899 TI - Regional distribution of sulfonylurea receptors in the brain of rodent and primate. AB - Glibenclamide, one of the most potent antidiabetic sulfonylureas, inhibits the activity of ATP-sensitive K+ channels in the pancreas as well as in the brain through its binding to specific receptors. Quantitative autoradiography was used to localize such receptors in the brain of rat, mouse, guinea-pig and marmoset, using [3H]glibenclamide as radioligand. In all four species, specific glibenclamide binding sites were found to be heterogeneously distributed. The highest densities were in the cerebral cortex, the molecular layer of the cerebellar cortex, the thalamus and the caudate-putamen. The globus pallidus and the substantia nigra were highly labelled in rat and mouse but poorly labelled in guinea-pig and marmoset. The distribution of glibenclamide binding sites in the hippocampus was different between the rodents and marmoset; in rodents, most binding sites were distributed in the fascia dentata and the CA3-CA4 fields of Ammon's horn, contrasting with a very homogeneous distribution in all subfields of the marmoset hippocampus. In conclusion, we demonstrate that primate brain contains specific binding sites for [3H]glibenclamide with a distribution not exactly similar to that in rodent brain. PMID- 8232900 TI - Chandelier cell axons identified by parvalbumin-immunoreactivity in the normal human temporal cortex and in Alzheimer's disease. AB - Parvalbumin is a calcium-binding protein which is thought to play a role in neuronal excitability. In the cerebral cortex parvalbumin is largely found in two subsets of GABAergic neurons, the chandelier and basket cells. A distinguishing characteristic of the chandelier cell is that the terminal portions of its axon form short vertical strings of boutons resembling candlesticks, which embrace the initial segment of pyramidal cell axon. In the present study, the terminals of chandelier cells in the human temporal cortex were immunostained with an antibody against parvalbumin. These terminals were found more abundantly in layers II and VI, less frequently in layers III and V, were hardly identified in layer IV, and absent in layer I. The relationship of parvalbumin-immunoreactive terminals and axon initial segments was further evidenced by re-sectioning identified rows of boutons into semithin sections. Electron microscopy of both temporal cortex and the somatosensory region of a biopsy sample revealed that these parvalbumin positive boutons indeed form symmetric synaptic contacts on the axon initial segments of pyramidal cells. As part of an enquiry into the possibility that these specialized interneurons may be involved in degenerative neurological diseases, the temporal lobes from seven patients with Alzheimer's disease were immunostained for parvalbumin. As in the control brains, the specific terminal portions of chandelier cells were recognized and identified in the temporal cortex by parvalbumin-immunocytochemistry. No major difference from normal brains was found, excepting for a lower density of candlesticks (30-35%) in layer II III. Since we showed in a previous study [Ferrer et al. (1991) J. neurol. Sci. 106, 135-141] that the number of parvalbumin-immunoreactive somata in the same Alzheimer's disease cases was not decreased, the observed reduction of terminals in layer II suggest that only the terminals of chandelier cells, but not the parent neurons, are decreased in Alzheimer's disease. PMID- 8232901 TI - Mutual interactions among cholinergic, noradrenergic and serotonergic neurons studied by ionophoresis of these transmitters in rat brainstem nuclei. AB - In urethane-anesthetized rats, single neuronal activity was recorded in or around the central gray of the caudal mesencephalon to rostral pons with multibarrel microelectrodes for ionophoretic application of acetylcholine, noradrenaline and serotonin. Neurons were classified by spike shape into broad-spike and brief spike neurons. In the laterodorsal tegmental nucleus, locus coeruleus or dorsal raphe, broad-spike neurons, marked by Pontamine Sky Blue and discriminated in sections processed for histochemistry of reduced nicotinamide adenine dinucleotide phosphate diaphorase or Nissl staining, were presumed to be cholinergic, noradrenergic or serotonergic, respectively. The majority of these neurons were inhibited through autoreceptors, except some laterodorsal tegmental neurons which might not be furnished by autoreceptors. Noradrenaline and serotonin inhibited more than two-thirds of the laterodorsal tegmental neurons tested, while a few neurons were excited by noradrenaline. Though effects of noradrenaline on dorsal raphe neurons and those of serotonin on locus coeruleus neurons were not clear in many neurons tested, neurons affected in these examinations (30%) were all inhibited clearly and no excitatory effect was observed. Acetylcholine exerted inhibition on about one-half of dorsal raphe neurons, while effects of acetylcholine on locus coeruleus neurons were the only case in the present study in which excitation was the major effect, though more than a half of locus coeruleus neurons were not sensitive to this drug. Thus, in this study some new data on the pharmacological properties of the cholinergic laterodorsal tegmental neurons were obtained. In addition, mutual interactions between brainstem cholinergic, noradrenergic and serotonergic neurons were assayed by comparing the pharmacological properties of these neurons tested with a uniform procedure. The interactions between these diffuse projection neurons may be involved in neural mechanisms controlling vigilance, wakefulness and/or sleep. PMID- 8232902 TI - Early developmental changes in [3H]nicotine binding in the human brainstem. AB - Little is known about the developmental profile of nicotinic cholinergic receptors in the developing human brain, despite the potential importance of such information in understanding the pathogenesis of neurological abnormalities or increased risk for the sudden infant death syndrome in offspring exposed to nicotine in utero. In this study, we determined the distribution of [3H]nicotine binding in the developing human brainstem by quantitative tissue autoradiography. In midgestational fetuses, [3H]nicotine binding sites were heavily concentrated in tegmental nuclei related to cardiopulmonary integration, arousal, attention, rapid eye movement sleep, and somatic motor control. Over the last half of gestation, [3H]nicotine binding decreased 60-70% in the tegmental nuclei, with a significant difference in binding between midgestation and early infancy. In contrast, there was essentially no change in [3H]nicotine binding in the major cerebellar-relay nuclei (principal inferior olive and griseum pontis) between the same time-points. Tritium quenching by increasing lipid (myelin) content in tissue sections did not account for the decreases in [3H]nicotine binding in tegmental nuclei. Based upon the high levels of [3H]nicotine binding at midgestation, combined with experimental data demonstrating trophic properties for acetylcholine, we postulate that nAChRs a role in the development of the brainstem tegmentum during this period, and that once this role is fulfilled, nicotinic cholinergic binding decreases and remains low thereafter. Alternatively, nicotinic cholinergic receptors may be critical for other developmentally related functions and/or neurotransmission in the brainstem tegmentum at midgestation. The high levels of [3H]nicotine binding in the brainstem tegmentum at midgestation and its rapidly changing profile over late gestation further suggest that mid-to-late gestation is a developmental period during which this region is likely to be most vulnerable to the harmful effects of nicotine in maternal cigarette smoke. The baseline information provided in this study is potentially relevant towards understanding attention deficits and risk for the sudden infant death syndrome in offspring exposed to cigarette smoke in utero. PMID- 8232903 TI - Morphological and electrophysiological peculiarities of sensory bush-like receptors of the frog urinary bladder. AB - Results of synchronous morpho-electrophysiological experiments conducted on living bush-like receptors of the urinary bladder of a frog (Rana temporaria) and results of their ultrastructural and cytochemical studies are presented in this article. It is demonstrated that these endings are mechanical receptors reacting to tactile stimuli. Rapidly and slowly adapting units are registered. The slowly adapting receptors prevail (94%). Morpho-topographic peculiarities of the rapidly and slowly adapting forms are recorded at the light (optic) level. Different variants of sediment deposits at Ranvier nodes of the myelinated afferent fibers are demonstrated by means of a ferri-ferricyanide technique. In this connection a hypothesis is discussed concerning the existence in bush-like receptors of several generators of impulses converging on to a solitary fiber. The ultrastructure of receptor terminals is demonstrated. Their connection with tissue elements of the organ wall is realized with the help of collagen fibrils braiding the terminals with a dense irregular net. No connections are discovered between terminals and cells with the help of desmosomes. It is supposed that this very interrelation of the receptor terminals with the surrounding tissues ensures a predominance of the slowly adapting forms among them. PMID- 8232904 TI - Long-term changes in striatal opioid systems after 6-hydroxydopamine lesion of rat substantia nigra. AB - The effects of unilateral 6-hydroxydopamine lesion of the nigrostriatal pathway on striatal opioid peptides and receptors were determined at different time intervals, from three days up to 24 weeks, post-lesion. Mu, delta and kappa opioid binding site densities in the ipsilateral caudate-putamen were decreased by 25-50% in rats which exhibited a greater than 90% loss of dopamine uptake sites. Differentiation of radioligand binding to kappa1 and kappa2 subtypes demonstrated a selective loss of kappa2 sites post-lesion. The onset of significant 6-hydroxydopamine lesion-induced changes in striatal opioid binding sites was delayed with respect to the loss of dopamine uptake sites. Furthermore, maximal loss of dopamine uptake sites was apparent within seven days post-lesion, but not until two to four weeks for mu, delta and kappa sites. In animals which exhibited an incomplete loss of dopamine uptake sites (less than 80%) there was no significant change in opioid binding site density. Striatal proenkephalin and prodynorphin messenger RNA levels were increased and decreased, respectively, after complete 6-hydroxydopamine lesion. Modulation of peptide messenger RNA levels was apparent within seven days and was maintained up to 24 weeks post lesion. In contrast, proenkephalin and prodynorphin messenger RNA levels were unchanged in animals which exhibited an incomplete loss of striatal dopamine uptake sites. Taken together, these observations suggest that the majority of mu, delta and kappa2 opioid binding sites are localized on non-dopaminergic elements in the caudate-putamen, but that substantia nigra innervation plays a role in the control of striatal opioid receptor expression. The 6-hydroxydopamine lesion induced decreases in striatal opioid binding site density may, in part, be a function of agonist-induced receptor downregulation. Alternatively, both opioid receptor and peptide expression in the caudate-putamen may be directly, but independently, regulated by ventral mesencephalic neurons. PMID- 8232905 TI - Rapid changes in the frequency tuning of neurons in cat auditory cortex resulting from pure-tone-induced temporary threshold shift. AB - The response areas (frequency by intensity) of single neurons in primary auditory cortex of anesthetized cats were studied before and after temporary threshold shifts in cochlear sensitivity induced by an intense pure tone. Cochlear temporary threshold shift was monitored through the threshold of the gross auditory nerve compound action potential and in most cases involved a notch-like loss centered at the characteristic frequency of the unit under study. Only two neurons showed changes in response area that mirrored the changes at the auditory periphery. Most neurons (14) showed more complex changes involving both expansion and contraction of response areas. Expansion of response areas was indicated by lower thresholds at some frequencies and by the emergence of sensitivity to previously ineffective frequencies. A change was classified as contraction when the response area after the intense-tone exposure was smaller than would be expected by applying the profile of the temporary threshold shift to the initial response area. Contraction of both upper (high intensity) and lower boundaries of response areas was found; in the most extreme cases, neurons were totally unresponsive after the intense-tone exposure. The complexity of effects of temporary threshold shifts on the response areas of cortical neurons is likely to be related to mechanisms that normally determine the frequency response limits of these neurons. The response areas of cortical neurons are more complex than those of auditory nerve fibers, and are thought to reflect the integration of excitatory and inhibitory inputs. The variety of effects observed in this study are consistent with the excitatory and inhibitory components of the response area of a given neuron being differentially affected by the temporary threshold shift. PMID- 8232906 TI - Long-term increase in protein kinase C-gamma and muscarinic acetylcholine receptor expression in the cerebral cortex of amygdala-kindled rats--a quantitative immunocytochemical study. AB - Kindling is an animal model for epilepsy in which repeated application of an electrical stimulus to brain pathways results in an epileptic focus. The animal holds a permanent state of hyperexcitability to the stimulus for the rest of its life. Understanding the cellular and molecular processes underlying hyperexcitability could provide insight into epileptogenesis. Furthermore, it could elucidate cellular and molecular bases of synaptic plasticity in the central nervous system. In the present study the long-term effect of a kindled focus in the amygdala on the gamma-isoform of protein kinase C and the muscarinic cholinergic receptor as cellular messengers was evaluated in the cerebral cortex of rats. Following an average of 10 bilaterally generalized seizures kindling stimulation was terminated and rats were left undisturbed for approximately three months. Brains were processed by immunocytochemistry using monoclonal antibodies against protein kinase C-gamma and muscarinic cholinergic receptor protein. Digital image analysis of sections through the entire forebrain revealed an increase in optical density of both protein kinase C-gamma and the muscarinic cholinergic receptor in the piriform and entorhinal cortex of the hemisphere contralateral to the stimulation site in kindled rats. However, on the ipsilateral side no change was observed in comparison with electrode implanted nonkindled control rats. The observed increase in expression of muscarinic cholinergic receptor protein and a component of the phosphoinositide second messenger system (protein kinase C-gamma) located in specific areas of the cerebral cortex in kindled rats could serve as a basis for the permanent state of hyperexcitability in these rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232907 TI - Kynurenine pathway enzymes in a rat model of chronic epilepsy: immunohistochemical study of activated glial cells. AB - The kynurenine pathway metabolites quinolinic acid and kynurenic acid have been hypothetically linked to the occurrence of seizure phenomena. The present immunohistochemical study reports the activation of astrocytes containing three enzymes responsible for the metabolism of quinolinic acid and kynurenic acid in a rat model of chronic epilepsy. Rats received 90 min of patterned electrical stimulation through a bipolar electrode stereotaxically positioned in one hippocampus. This treatment induces non-convulsive limbic status epilepticus that leads to chronic, spontaneous, recurrent seizures. One month after the status epilepticus, the rats showed neuronal loss and gliosis in the piriform cortex, thalamus, and hippocampus, particularly on the side contralateral to the stimulation. Astrocytes containing the kynurenic acid biosynthetic enzyme (kynurenine aminotransferase) and the enzymes for the biosynthesis and degradation of quinolinic acid (3-hydroxyanthranilic acid oxygenase and quinolinic acid phosphoribosyltransferase, respectively) became highly hypertrophied in brain areas where neurodegeneration occurred. Detailed qualitative and quantitative analyses were performed in the hippocampus. In CA1 and CA3 regions, the immunostained surface area of reactive astrocytes increased up to five-fold as compared to controls. Enlarged cells containing the three enzymes were mainly observed in the stratum radiatum, whereas the stratum pyramidale, in which neuronal somata degenerated, showed relatively fewer reactive glial cells. Hypertrophied kynurenine aminotransferase- and 3 hydroxyanthranilic acid oxygenase-immunoreactive cells were comparable in their morphology and distribution pattern. In contrast, reactive quinolinic acid phosphoribosyl transferase-positive glial cells displayed diversified sizes and shapes. Some very large quinolinic acid phosphoribosyl transferase-immunoreactive cells were noticed in the molecular layer of the dentate gyrus. In the hippocampus, the number of immunoreactive glial cells increased in parallel to the hypertrophic responses. In addition, pronounced increases in immunoreactivities, associated with hypertrophied astrocytes, occurred around lesioned sites in the thalamus and piriform cortex. These findings indicate that kynurenine metabolites derived from glial cells may play a role in chronic epileptogenesis. PMID- 8232908 TI - Electrophysiological properties of intralaminar thalamocortical cells discharging rhythmic (approximately 40 HZ) spike-bursts at approximately 1000 HZ during waking and rapid eye movement sleep. AB - Thalamocortical neurons located in the large-celled district of the cat intralaminar centrolateral nucleus were found to discharge spike-bursts with unusually high frequencies (800-1000 Hz) during spindle oscillations of the electroencephalogram. In chronically implanted animals, similar spike-bursts were also fired during wakefulness and rapid eye movement sleep, two behavioral states in which other thalamocortical neurons tonically fire single spikes. Such high frequency spike-bursts recurred with a fast rhythm of 20-40 Hz during waking and rapid eye movement sleep. Intracellular recordings under barbiturate anesthesia showed that, during spindle oscillations, the spike-bursts of intralaminar neurons are generated by brief low-threshold spikes with a much shorter refractory phase than in other thalamocortical cells. Depolarizing pulses from the resting membrane potential triggered fast oscillations (20-80 Hz) crowned by short high-frequency (800-1000 Hz) spike-bursts. During the inter-spindle epochs, the "tonic" firing of these neurons was, in fact, a fast oscillation (30-40 Hz) of the membrane potential leading to single spikes or spike-doublets. Autocorrelograms computed from inter-spindle epochs, at relatively depolarized levels, confirmed the presence of multiple peaks at this fast rhythm. The properties of these neurons make them well suited for the distribution of fast rhythms during arousal and rapid eye movement sleep over the cerebral cortex. PMID- 8232909 TI - Functional connectivity from CA3 to the ipsilateral and contralateral CA1 in the rat dorsal hippocampus. AB - Anatomical advances have led to a reappraisal of the organization of hippocampal circuitry. However, it is not clear whether the functional connectivity is fully determined by the anatomical connectivity or whether it is significantly modified by feed-forward inhibition and modulatory inputs. Therefore, we have mapped CA1 responses evoked by stimulation of ipsilateral and contralateral CA3 in vivo. Population spike amplitude and threshold were plotted to produce response maps. All CA3 subregions projected diffusely to ipsilateral CA1. However, a pattern of maximal response emerged. Caudal CA3 stimulation evoked the maximal responses septally, while rostral CA3 responses were maximal temporally. The ipsilateral CA3 response maps were compared with those produced by stimulation at the homotopic point in the contralateral CA3. The CA1 areas of maximal functional connectivity were the same implying that there is convergence of the input to CA1 from homotopic CA3 sites in the two hippocampi. Although a response in CA1 was evoked widely, our results suggest that the functional connectivity is ordered, within and between the dorsal hippocampi, and that it is consistent with the recent anatomical data. The present findings allow more precise study of the propagation of normal and abnormal neuronal activity, within and between the dorsal hippocampi. Information on the site and speed of propagation of neuronal activity would be necessary for the development of a physiologically realistic model of hippocampal computation. PMID- 8232910 TI - The distribution of Alz-50 immunoreactivity in the normal human brain. AB - Alz-50 is a monoclonal antibody that recognizes normal tau proteins as well as phosphorylated tau proteins that are associated with paired helical filaments in Alzheimer's disease. To establish an accurate baseline for future pathological studies, we examined the distribution of Alz-50 immunoreactivity in normal human brain from infancy to senescence. We found extensive staining patterns of somata and axonal profiles in the striatum, amygdala, hypothalamus, brainstem and spinal cord in all normals at all ages. Similar normal staining patterns were seen in the brains of patients who had suffered trauma, tumors, cerebral infarcts, grade 1 periventricular hemorrhages, and in those who had suffered from amyotrophic lateral sclerosis, Parkinson's disease, multi-systems atrophy and Shy-Drager syndrome. An absence of cell body staining and only minimal axonal staining was noted in the same brains with immunocytochemistry using PHF-1, a monoclonal antibody generated against paired helical filament proteins from Alzheimer brains. The characteristic staining pattern of Alz-50 in normal brains is substantially more extensive than has previously been recognized. This pattern, which presumably describes a specific class of tau proteins, must be distinguished from the pathological staining observed in neurodegenerative diseases. PMID- 8232911 TI - gamma-Hydroxybutyric acid induced spike and wave discharges in rats: relation to high-affinity [3H]gamma-hydroxybutyric acid binding sites in the thalamus and cortex. AB - gamma-Hydroxybutyric acid is a naturally occurring compound which induces bilaterally synchronous spike and wave discharges in rats. The gamma hydroxybutyric acid model of absence seizures simulates clinical absence seizures behaviorally as well as electrographically. The present study was undertaken in order to establish the role of the high-affinity gamma-hydroxybutyric acid binding sites in the generation of gamma-hydroxybutyric acid-induced spike and wave discharges. Spike and wave discharges induced by gamma-hydroxybutyric acid were recorded with the aid of bipolar depth electrodes implanted in discrete regions of thalamus, cortex and hippocampus. In the present study we found that ventroposterolateral, ventroposteromedial, medial and the reticular nuclei of the thalamus discharged synchronously with the cortical generation of spike and wave discharges. In the cortex, the superficial layers (I-IV) of frontoparietal cortex generated spike and wave discharges, whereas no spike and wave discharges were recorded from deeper layers (V-VI) of frontoparietal cortex. At the onset of spike and wave discharges induced by gamma-hydroxybutyric acid, a rapid but reversible upregulation of gamma-hydroxybutyric acid binding sites was observed. This increased [3H]gamma-hydroxybutyric acid binding was characterized by an increase in the number of gamma-hydroxybutyric acid sites with no significant change in their affinity for gamma-hydroxybutyric acid. Moreover, the change in [3H]gamma-hydroxybutyric acid binding was observed only in those thalamic structures and cortical layers which were found to be involved in the generation of spike and wave discharges induced by gamma-hydroxybutyric acid. The CA3 field or dorsal hippocampus possesses the highest density of [3H]gamma-hydroxybutyric acid binding sites of all brain regions. However, no significant change in [3H]gamma-hydroxybutyric acid binding was observed in this region nor was the CA3 field involved in the generation of spike and wave discharges during gamma hydroxybutyric acid-induced absence-like seizures. These findings confirm that gamma-hydroxybutyric acid-induced absence-like seizures originate from thalamocortical pathways and that the onset of gamma-hydroxybutyric acid-induced spike and wave discharges is directly related to the regulation of gamma hydroxybutyric acid binding sites in those regions which constitute the involved thalamocortical loop. PMID- 8232912 TI - Compartmentation of the cerebellar cortex by protein kinase C delta. AB - Six isozymes of protein kinase C were analysed in the rat cerebellum using immunohistochemistry. The results revealed a non-uniform distribution of protein kinase C delta among Purkinje, basket, and stellate cells. Serial-section mapping of the delta immunoreactivity revealed that (i) the number and intensity of labeled Purkinje cells increased from rostral to caudal while labeled basket stellate cells decreased caudally; (ii) the majority of Purkinje cells were labeled in the nodulus, flocculus, and paraflocculus while the anterior lobules were mostly negative; and (iii) labeled Purkinje cells formed distinct parasagittal bands in lobules 6-9 of the paravermis and vermis. The banding of protein kinase C delta within subsets of Purkinje cells suggests units of cerebellar circuitry with specific signaling properties through protein phosphorylation. The visual-vestibular regions of the cerebellum contained the highest amount of the isozyme. PMID- 8232913 TI - Cobalt uptake enables identification of capsaicin- and bradykinin-sensitive subpopulations of rat dorsal root ganglion cells in vitro. AB - A novel modification of the stimulated cobalt uptake technique has been used to identify rat dorsal root ganglion cells expressing capsaicin and bradykinin receptors. The technique involves incubating intact dorsal root ganglia in vitro in a modified Krebs solution in which cobalt chloride has been substituted for calcium. Activation of dorsal root ganglion cells by capsaicin or bradykinin in the presence of the cobalt ions results in cobalt influx into the excited cells. Histochemical methods were then used to visualize the intracellular accumulation of cobalt, and labelled cells were counted and characterized. Capsaicin (2 microM) or bradykinin (500 nM) applied for 20 min induced cobalt uptake in 13.8 +/- 0.6 and 9.6 +/- 0.5% of neuronal profiles in dorsal root ganglia (L4), respectively, a significantly larger number than stained in control ganglia (in the absence of agonists: 1.8 +/- 0.7%). The longest diameter of the soma of stained dorsal root ganglion cells following capsaicin and bradykinin perfusion were significantly different from each other and from the non-labelled population (17.5 +/- 0.7 and 24.5 +/- 0.2 microns for capsaicin; 23.2 +/- 0.9 and 25.5 +/- 0.4 microns for bradykinin; labelled and non-labelled cells, respectively). The distribution of cell diameters revealed that while capsaicin-sensitive cells were exclusively small-sized, bradykinin-sensitive cells were predominantly small and medium sized. The selective bradykinin-2 receptor antagonist HOE-140 (5.0 microM) blocked the bradykinin-induced staining (2.16 +/- 0.02%) but not that of capsaicin. The bradykinin-1 agonist [des-Arg9]-bradykinin did not induce any significant increase in stained cells over the control number (2.2 +/- 0.7%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232914 TI - What do the synaptic vesicles contain? AB - The intersynaptic membranes of the rat brain cortex were found to remain firmly attached to one another after perfusion of strongly anisotonic solutions. Brains perfused with depolarizing and excitotoxic agents showed abundant, apparent intermingling of mitochondria and synaptic vesicles. The results suggest (i) that the intersynaptic membranes are not separated from one another by an essentially fluid intersynaptic medium as it is commonly assumed, but rather firmly attached to one another by a layer of faintly osmiophilic yet remarkably stable, water insoluble material; and (ii) that the synaptic vesicles may be involved in adenosine triphosphate carriage. Well established multidisciplinary data are presented which appear to be in line with both possibilities. PMID- 8232915 TI - Restoration of complex sensorimotor behavior and skilled forelimb use by a modified nigral cell suspension transplantation approach in the rat Parkinson model. AB - While intrastriatal transplants of dopamine-rich ventral mesencephalic tissue are effective in reversing a variety of drug-induced behaviors in the rat Parkinson model, previous studies have failed to obtain significant graft-induced effects on deficits in certain aspects of complex sensorimotor behaviors. In the present study we have applied a modified cell suspension transplantation procedure, which allows more reproducible and consistent ventral mesencephalic transplants of large size, as well as more wide-spread distribution of the ventral mesencephalic tissue over multiple graft sites within the denervated caudate-putamen. Using this approach it has for the first time been possible to obtain significant amelioration of the lesion-induced deficits in skilled forelimb use and in the rats ability to switch from one behavior (eating) to another (orientation towards tactile stimuli), so-called disengage behavior. Rats with unilateral 6 hydroxydopamine lesions of the mesostriatal dopamine pathway received a total of 450,000 fetal ventral mesencephalic cells, implanted either as two large deposits along a single injection tract ("Macro" grafts), or as 18 small deposits along six injection tracts in the head of the denervated caudate-putamen ("Micro" grafts) and the behavioral changes were studied up to three months after transplantation. On the drug-induced tests, both types of transplants reversed amphetamine- and D1-receptor agonist-induced turning, and produced a partial (50 75%) reduction in apomorphine-induced and D2-receptor agonist-induced turning. On the spontaneous sensorimotor tests, both types of grafts reversed the deficit in simple sensorimotor orientation. In addition, the Micro-grafted animals (which produced the most extensive reinnervation of the denervated striatum) showed a significant improvement in skilled forelimb use and in response latency in the disengage behavior test. Although the large sized Macro-grafted animals showed a similar trend, it did not reach significance. Moreover, the Micro grafts had a more pronounced effect on spontaneous turning behavior in a conditioned response test. The improvement in response latency in the disengage test was significantly correlated with the dopamine level in the nucleus accumbens, whereas the magnitude of the conditioned turning response was significantly correlated with the dopamine levels in the head of the caudate-putamen. The results show that intrastriatal nigral transplants, despite their ectopic placement, can ameliorate lesion-induced deficits also in more complex sensorimotor behaviors. This improved graft effect is likely to depend on both extensive dopaminergic reinnervation throughout the head of the caudate-putamen, as well as on closer integration of the grafted nigral tissue with the host striatal circuitry. PMID- 8232916 TI - A fetal nigral graft prevents behavioral supersensitivity associated with repeated injections of L-dopa in 6-OHDA rats. Correlation with D1 and D2 receptors. AB - The effect of repeated administration of L-3,4-dihydroxyphenylalanine was studied behaviorally and biochemically in grafted versus non-grafted rats with a 6 hydroxydopamine unilateral lesion of the dopaminergic nigro-striatal pathway. Non grafted rats receiving 14 injections of L-3,4-dihydroxyphenylalanine increased their contraversive circling while grafted rats did not, even after fourteen injections. The density of striatal dopamine receptors was examined by autoradiography using the ligands [3H]-SCH 23390 for dopamine D1 receptors and [3H]-spiperone for D2 receptors. In rats with a lesion of the nigro-striatal dopaminergic pathway, an increase of [3H]-SCH 23390 and [3H]-spiperone binding in the lesioned striatum was observed when compared with the striatum on the intact side. Chronic treatment with L-3,4-dihydroxyphenylalanine led to a further increase in D1 receptor density in the lesioned as well as the intact side. A similar pattern was observed for D2 receptors although the change did not reach significance. A graft of fetal nigral neurons brought the density of both D1 and D2 receptors on the lesioned side back to the level of the intact side. This is observed both in acutely or chronically L-3,4-dihydroxyphenylalanine treated rats. This study suggests that nigral grafts protect the striatum against L-3,4 dihydroxyphenylalanine-induced supersensitivity. It appears that the graft preserves the symmetry of the striatum even though there is an increase of D1 dopamine receptors. These results suggest that a fetal nigral graft could prevent the induction of 3-4-dihydroxyphenylalanine- induced dyskinesia in parkinsonian patients. PMID- 8232917 TI - Direct interaction with target-derived glia enhances survival but not differentiation of human fetal mesencephalic dopaminergic neurons. AB - Regulation of the developing nervous system involves attraction, guidance and modification of innervating neurons by target cells through diffusible and membrane-related factors. The trophic effects from specific cell types remain to be investigated and characterized. In a series of experiments in which human fetal mesencephalic dopaminergic cells were co-cultured with target or non-target neurons or glial cells in direct or contiguous contact, we demonstrate that striatal glial cells (target-derived glia) can enhance dopaminergic neuron survival by up to 400% compared to either non-target cell co-cultures or mesencephalic controls. When in direct contact with striatal neurons, a greater proportion of dopaminergic neurons had a more differentiated morphology. The enhancement of dopaminergic neuron survival by target-derived glia appears to be mediated both by direct contact, possibly through target membrane-specific phenomena, and by diffusible substances, whereas non-target glia appear to exert the trophic effects predominantly through the latter mechanism. The finding that target neurons influence mainly dopaminergic neuron differentiation and target glia their survival indicates multiple, target cell type-specific regulation of innervating neuron development. These findings also have relevance to the establishment of neuronal cultures for neural transplantation. PMID- 8232918 TI - Basal forebrain stimulation facilitates tone-evoked responses in the auditory cortex of awake rat. AB - The effects of unilateral basal forebrain stimulation on the tone-evoked responses recorded in the auditory cortex ipsilateral and contralateral to the stimulation site, were investigated in fully awake rats. After 10 tone alone presentations, 20 pairing trials were given during which the basal forebrain stimulation was followed by the tone 30 ms later. Ten test-tones were presented immediately, 15 min and 1 h after pairing. Immediately after pairing, the short latency "on" and "off" tone-evoked responses were enhanced in the ipsilateral but not in the contralateral cortex. This enhancement did not persist 15 min later. Systemic atropine injection prevented the ipsilateral facilitation. The responses to the tone were not modified when tested after 20 basal forebrain stimulations delivered in the absence of the tone. These results are the first demonstration in awake animals that an activation of the auditory cortex by cholinergic neurons of the basal forebrain is able to facilitate cortical responsiveness. A temporal contiguity between the cholinergic activation and the neuronal discharges elicited by the sensory stimulus is required for the facilitation to take place. The results are compared to previous ones obtained in anesthetized animals, and the functional role of cholinergic activation from the basal forebrain in cortical processing is discussed. PMID- 8232919 TI - Decreased density of forebrain cholinergic neurons in experimental autoimmune dementia. AB - Sera of Alzheimer's disease and Down's syndrome patients contain antibodies which bind specifically to the high molecular weight neurofilament protein of Torpedo cholinergic neurons. We have recently shown that prolonged immunization of rats with this antigen results in the accumulation of IgG in neurons in the septum and hippocampus of the immunized rats and in cognitive impairments. This animal model is termed experimental autoimmune dementia. In the present study we examined whether the anti-cholinergic high molecular weight neurofilament subunit immune response of the experimental autoimmune dementia rats affects forebrain cholinergic neurons. This was performed immunohistochemically utilizing a monoclonal antibody to nerve growth factor receptor, a specific marker of cholinergic neurons in the forebrain. The results obtained revealed significant decreases in the density of cholinergic neurons in the medial septal nucleus and diagonal band of the experimental autoimmune dementia rats. These decreases are specific to the anti-cholinergic high molecular weight neurofilament subunit immune response of the experimental autoimmune dementia rats and are not observed in control rats which were immunized with chemically heterogeneous high molecular weight neurofilament subunit. The decrease in density of forebrain cholinergic neurons in experimental autoimmune dementia rats may mimic pathogenic processes in Alzheimer's disease and supports a role for anti-cholinergic high molecular weight neurofilament subunit antibodies in the degeneration of cholinergic neurons in the disease. PMID- 8232920 TI - Disintegration of the spatial organization of behavior in experimental autoimmune dementia. AB - Experimental autoimmune dementia is a rat model designed to examine the potential role of anti-cholinergic neurons antibodies in neuronal degeneration in dementia and Alzheimer's disease. We have previously shown that sera of patients with Alzheimer's disease contain antibodies which bind specifically to the high molecular weight neurofilament protein of the purely cholinergic electromotor neurons of Torpedo. Production of such antibodies in experimental autoimmune dementia rats by prolonged immunization with the Torpedo cholinergic high molecular weight neurofilament subunit results in accumulation of antibodies in the septum and hippocampus of the immunized rats, in a marked decrease in the density of forebrain cholinergic neurons, and in memory deficits. In the present study we characterized the open-field behavior of experimental autoimmune dementia rats, and examined whether, like in dementia, the spatiotemporal organization of their behavior is impaired. The results obtained revealed that experimental autoimmune dementia rats travel shorter distances; explore a smaller part of the open-field; and perform less round-trips to the key location--the home base--in reference to which their behavior is normally organized. The shrinkage of the explored space and the reduced number of round trips are independent of the amount of locomotion and represent a deterioration in the organization of behavior in time and space. These behavioral changes are specific to the anti-cholinergic immune response of experimental autoimmune dementia rats as they are not observed in rats which were immunized with chemically heterogeneous high molecular weight neurofilament subunit.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8232921 TI - Clinical trials in stroke prevention--persisting uncertainties, firm answers. PMID- 8232922 TI - The neurologic syndrome of vitamin E deficiency: a significant cause of ataxia. PMID- 8232923 TI - Mitochondrial involvement in Parkinson's disease: the controversy continues. PMID- 8232924 TI - Death after death: the presumption of informed consent for cardiopulmonary resuscitation--ethical paradox and clinical conundrum. PMID- 8232925 TI - Friedreich's ataxia phenotype not linked to chromosome 9 and associated with selective autosomal recessive vitamin E deficiency in two inbred Tunisian families. AB - Friedreich's ataxia (FA) is an autosomal recessive neurodegenerative disorder, the disease locus (FRDA) of which has been assigned to 9q13-q21.1 by genetic linkage analysis in affected families. We report two large inbred Tunisian families with FA manifestations that did not show the expected linkage. The disease locus could be excluded from a large (12 cMo) region around FRDA. This is the first report providing evidence for nonallelic genetic heterogeneity for the FA clinical phenotype. On subsequent analysis, all patients had very low levels of serum vitamin E whereas the parents and healthy sibs had normal vitamin E levels. This establishes that the selective vitamin E deficiency with normal fat absorption is an autosomal recessive trait, which is associated in the two families reported here with the FA phenotype. PMID- 8232926 TI - Subtle extrapyramidal signs can predict the development of dementia in elderly individuals. AB - To assess the significance of mild extrapyramidal signs (EPSs) in aging, we prospectively followed 228 elderly individuals without dementia or major neurologic disease at baseline. Seventy-three (32%) showed mild idiopathic EPSs at baseline assessment and 32 (14%) of these 228 individuals met NINCDS-ADRDA criteria for probable or possible Alzheimer's disease on follow-up. While none of these individuals developed Parkinson's disease, those with more than one EPS at baseline were significantly more likely to become demented during follow-up than were those with one or no EPS, an association not confounded by age, education, or gender. Individuals with more than one EPS and with cognitive impairment (CI) at baseline were at highest risk for dementia. EPSs were also independently associated with incident dementia in the absence of CI at baseline. We conclude that mild EPSs are a significant predictor of dementia in elderly individuals. PMID- 8232927 TI - Can sensory stimulation improve the functional outcome in stroke patients? AB - After obtaining informed consent, we randomized 78 patients with severe hemiparesis of the left or right side within 10 days of stroke onset: 40 to a control group receiving daily physiotherapy and occupational therapy, and 38 to a group that, in addition, we treated with sensory stimulation (acupuncture) twice a week for 10 weeks. The median age was 76 years for both groups. Motor function, balance, and ADL (Barthel's Index) were assessed before the start of treatment and at 1 and 3 months after stroke onset; ADL was also assessed after 12 months. We assessed the quality of life (QL) using the Nottingham Health Profile 3, 6, and 12 months after stroke onset. Patients given sensory stimulation recovered faster and to a larger extent than the controls, with a significant difference for balance, mobility, ADL, QL, and days spent at hospitals/nursing homes. Whether acupuncture per se is responsible for the differences requires further study. PMID- 8232928 TI - Pontine versus capsular pure motor hemiparesis. AB - We prospectively studied 21 patients with pure motor hemiparesis (PMH). CT showed a capsular lesion consistent with the clinical syndrome in 15 patients (71%) and was repeatedly negative in the remaining six (29%). In all six patients with repeatedly negative CT, MRI showed a pontine paramedian infarct as the notable cause of PMH. Clinical findings could not definitely distinguish between capsular and pontine PMH, but the combination of dysarthria and a history of previous transient gait abnormality or vertigo favored a pontine location. Outcome at 3 months was characterized by persistent, moderate to severe disability in 86% of patients with pontine PMH versus 46% in capsular PMH. Based on MRI and magnetic resonance angiographic findings, the presumed mechanism of pontine ischemic lesions was a lacunar process in most instances (86%). PMID- 8232929 TI - Presentation and initial clinical course in patients with chronic inflammatory demyelinating polyradiculoneuropathy: comparison of patients without and with monoclonal gammopathy. AB - Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) may occur in association with a monoclonal gammopathy of undetermined significance (MGUS) or a variety of other systemic illnesses. It is not known if the clinical features of CIDP are altered by the presence of an MGUS. We compared demographic features, clinical presentation, improvement and outcome after initial treatment, and electrodiagnostic features of a group of 77 patients with idiopathic CIDP (CIDP I, no associated systemic illness) with 26 patients with CIDP in whom an MGUS was found during evaluation of the neuropathy (CIDP-MGUS). Patients with CIDP-MGUS had, on average, a more indolent course and less severe weakness than patients with CIDP-I, despite similar motor conduction studies. CIDP-MGUS patients also demonstrated less functional impairment, more frequent sensory loss, and more abnormal sensory conduction studies than patients with CIDP-I. Because of the greater improvement of CIDP-I patients with treatment, both groups had similar outcomes from their initial episodes of weakness. Subgroup analysis of CIDP-MGUS patients did not demonstrate differences between groups with IgM and IgG or IgA gammopathies. PMID- 8232930 TI - Seronegativity for type 1 antineuronal nuclear antibodies ('anti-Hu') in subacute sensory neuronopathy patients without cancer. AB - We followed 21 patients with sensory neuronopathy without evidence of cancer for up to 23 years. All were seronegative for type 1 antineuronal nuclear antibodies (ANNA-1, also called "anti-Hu"). We additionally studied 67 seropositive patients with sensory neuropathy or a related neurologic syndrome. Ninety-one percent of the seropositive patients had a small-cell lung carcinoma. One, with a normal chest x-ray, had been followed for 7 years for sensory neuronopathy of indeterminate cause before serologic testing for ANNA-1 led to the discovery of the tumor by CT. We conclude that ANNA-1 seropositivity in a patient with sensory neuronopathy is strong evidence for an underlying small-cell lung cancer. PMID- 8232931 TI - Familial essential tremor and idiopathic torsion dystonia are different genetic entities. AB - Familial essential tremor (ET) is an autosomal dominant disorder presenting as an isolated postural tremor. Its frequent association with dystonia suggests that the two disorders might be pathogenically related. We report the exclusion of the DYT1 locus on chromosome 9q32-34, responsible for idiopathic torsion dystonia (ITD), in two large ET families. We conclude that ET and ITD are distinct genetic disorders. PMID- 8232932 TI - Total body irradiation for myasthenia gravis: a long-term follow-up. AB - Total body irradiation (TBI) produces prolonged immunosuppression with rare side effects. We studied 12 thymectomized patients affected with chronic generalized severe myasthenia gravis. All patients had been totally or partially refractory to prolonged oral treatment with immunosuppressive drugs, and most had contra indications for these drugs. Low-dose (1.8- to 2.3-Gy total dose) TBI was administered in single, 0.1-Gy doses, two to three times per week. TBI was well tolerated and was associated with objective clinical improvement in six patients, lasting more than 2 years in five. In addition, TBI produced a long-lasting lymphopenia with a pronounced decrease of T CD4+ lymphocytes; T CD8+ lymphocytes were almost unchanged over the 2 years of the study. CD16+ and CD20+ lymphocytes, after an initial decrease, increased above baseline. TBI was also associated with decreased anti-AChR antibody titer. The decrease of lymphocyte count and of anti AChR antibody titer was more pronounced in the patients who improved, suggesting that lymphopenia and immunosuppression may have contributed to clinical improvement. PMID- 8232933 TI - Autosomal dominant parkinsonism with benign course and typical Lewy-body pathology. AB - The few previously reported patients with familial parkinsonism and Lewy-body pathology in the substantia nigra displayed a variety of clinical and pathologic syndromes. We now describe a family with very slowly progressive Parkinson's disease (PD) that has, in most cases, responded poorly to levodopa and includes subjective visual difficult. Four personally confirmed cases--with onset at ages 35, 25, 16, and 16-have occurred in three generations, and four suspicious cases have occurred in three other generations. There has been a trend toward progressively younger age of onset. One autopsied case showed a distribution of cell loss and Lewy bodies typical of PD. The hereditary pattern is most compatible with autosomal dominance. This kindred's illness shows that a presumably single Mendelian dominant gene can cause the clinical and pathologic features of PD, and further extends the clinical spectrum of pathologically typical Lewy-body PD. PMID- 8232934 TI - Risk factors for nursing home placement in advanced Parkinson's disease. AB - We evaluated the risk factors for nursing home placement of patients with Parkinson's disease (PD) by matching 11 PD patients permanently admitted to nursing homes with two control PD patients remaining at home. Risk factors assessed were motor severity, presence of hallucinations/delusions, and presence of memory of problems. The only statistically significant risk factor was the presence of hallucinations/delusions. Motor severity and the presence of memory problems did not have an impact on nursing home placement. There was no risk factor synergy for hallucinations, motor disability, and mental impairment. Since all patients in this series who entered nursing homes remained there permanently, these data suggest that vigorous efforts to control hallucinations may be warranted to prevent nursing home placement. PMID- 8232935 TI - Clinical-neuropathologic correlation in HIV-associated dementia. AB - The structural abnormalities that correlate with the clinical manifestations of HIV-associated dementia (HIVD) are unclear. In a prospectively categorized group of patients with and without HIVD who were followed to autopsy, we correlated HIV related neuropathologic changes with the presence and severity of HIVD. We also assessed the effect of antiretroviral therapy on the neuropathologic changes. Finally, using reverse transcriptase-polymerase chain reaction on homogenized brain tissue, we correlated the relative expression of mRNA for tumor necrosis factor-alpha (TNF-alpha) with cognitive impairment and with the patterns of neuropathologic changes. The presence of multinucleated giant cells and diffuse myelin pallor were specific for HIVD, but these pathologic changes occurred in only 50% of patients with dementia. Patients treated with antiretroviral agents for > 12 months were less likely to show multinucleated giant cells or diffuse myelin pallor. Levels of mRNA for TNF-alpha from frontal subcortical white matter were significantly greater in patients with HIVD than in AIDS patients without dementia or in seronegative controls. We conclude that routine histopathologic examination of the brain fails to detect multinucleated giant cells and diffuse myelin pallor in 50% of patients dying with HIVD. This suggests that more subtle neuropathologic correlates for the clinical manifestations of HIVD exist. Our observations of elevated levels of TNF-alpha mRNA in HIVD indicate that indirect mechanisms of brain dysfunction, such as abnormal cytokine expression, may contribute to the pathogenesis of HIVD. PMID- 8232936 TI - Oblique misdirection and slowing of vertical saccades after unilateral lesions of the pontine tegmentum. AB - Three patients with unilateral lesions of the pontine tegmentum, identified by CT and MRI, had abnormal vertical saccades and slowed ipsilateral horizontal saccades. Attempted vertical saccades were misdirected obliquely, away from the side of the lesion, and their vertical components were prolonged. Oblique saccades had curved trajectories and prolonged durations of their vertical components. Unilateral damage to excitatory burst neurons and pause cells in the medial part of the caudal paramedian pontine reticular formation may cause these abnormal vertical and oblique saccades. Misdirection and slowing of vertical saccades can accompany the paralysis or slowing of ipsilateral horizontal saccades caused by pontine damage. PMID- 8232937 TI - Dementia in AIDS patients: incidence and risk factors. Multicenter AIDS Cohort Study. AB - We determined incidence and future projections of dementia after AIDS onset in 492 homosexual men with AIDS in the Baltimore/Los Angeles sites of the Multicenter AIDS Cohort Study, 64 of whom developed dementia. We studied various risk factors for dementia, including demographic and clinical features, medical history, markers of immune status before AIDS, and zidovudine use. During the first 2 years after AIDS, HIV dementia developed at an annual rate of 7%. Overall, 15% of the cohort followed through death developed dementia. The median survival after dementia was 6.0 months. Using a proportional hazards model, risk factors for more rapid development of dementia were lower hemoglobin (relative hazard, 0.59 per additional 2 g/dl; p = 0.0005) and body mass index (relative hazard, 0.64 per additional 5 kg/m2; p = 0.05) 1 to 6 months before AIDS, more constitutional symptoms 7 to 12 months before AIDS (relative hazard, 1.68 per additional symptom, p = 0.005), and older age at AIDS onset (relative hazard, 1.60 per decade older; p = 0.009). In a multivariate model, pre-AIDS hemoglobin remained the most significant predictor of dementia. There were no significant risks defined from demographic characteristics, specific AIDS-defining illnesses, zidovudine use before AIDS, or CD4+ lymphocyte count before AIDS. We project that 12 months after the first AIDS diagnosis, 7.1% of survivors will have dementia. The observed association between anemia, low weight, constitutional symptoms, and dementia suggests a role for cytokines inducing both systemic and neurologic disease. PMID- 8232938 TI - Abnormal cortical responses in patients with writer's cramp. AB - We evaluated sensorimotor processing in patients with writer's cramp using PET and H2(15)O blood flow scans. The study included six right-handed patients with unilateral writer's cramp and eight right-handed normals. Subjects had blood flow scans at rest and during vibration of either the "affected" or "unaffected" hand. Vibration produced a consistent peak response in primary sensorimotor area (PSA) and supplementary motor area (SMA), both contralateral to the vibrated hand. Both responses were significantly reduced approximately 25% in patients with writer's cramp (PSA, p = 0.002; SMA, p = 0.02) whether vibrating the affected or unaffected hand. This indicates that patients with unilateral writer's cramp have bilateral brain dysfunction. These data provide objective evidence of abnormal central sensorimotor processing in writer's cramp. PMID- 8232939 TI - Mitochondrial respiratory chain activity in skeletal muscle from patients with Parkinson's disease. AB - Different abnormalities in mitochondrial electron transport chain activity have been demonstrated in muscle and other tissues of patients with idiopathic Parkinson's disease (PD). We studied eight Spanish patients with PD to evaluate the functional activity of the electron transport chain in muscle mitochondria from patients of this country. We found lower complex I activity (nmol.min-1.mg 1) in patients (245.8 +/- 42.8) than in controls (331.6 +/- 60.1) (p = 0.004) and lower complex IV activity in patients (46.1 +/- 9) than in controls (144.1 +/- 42.3) (p = 0.00001). Complex V activity was also decreased in two patients and complex II and III activities were normal in all of them. Although these results strongly suggest an alteration in mitochondrial DNA in PD, the various electron transport chain defects in different tissues seem to be nonspecific. PMID- 8232940 TI - Respiratory chain and mitochondrial DNA in muscle and brain in Parkinson's disease patients. AB - There are several reports of a defect of complex I in the substantia nigra (SN) of Parkinson's disease (PD) patients. To evaluate whether this is specific to dopaminergic neurons or the phenotypically relevant consequence of a widespread failure of the mitochondrial oxidative phosphorylation (OXPHOS) system, we measured respiratory enzyme activities in muscle homogenates from 16 PD patients and eight age-matched controls, and in muscle isolated mitochondria of six PD patients and six age-matched controls. We found no difference between the PD and control groups. In addition, we detected, by polymerase chain reaction, the mitochondrial DNA (mtDNA) "common deletion" (CD) in muscle specimens of 14 of 17 PD patients, but we obtained similar results in age-matched controls. In both groups, the amount of CD-specific deleted (delta) mtDNA ranged from 0.0% to 0.1%. Our data suggest that PD cannot be attributed to a multisystem decline of mitochondrial OXPHOS, and that lesions of muscle mtDNA in PD are likely due to normal aging. However, there was a remarkable accumulation of delta mtDNA in the SN of a PD patient and an age-matched control, suggesting that the SN is exquisitely sensitive to age-dependent damage of the mitochondrial genome. PMID- 8232941 TI - N24 spinal response to tibial nerve stimulation and magnetic resonance imaging in lesions of the lumbosacral spinal cord. AB - We studied tibial nerve somatosensory evoked potentials (SEPs) in 22 patients with progressive clinical signs of sensory and motor involvement of the lower limbs. Magnetic resonance imaging (MRI) in 21 patients revealed lesions of the thoracolumbar cord in 18 and of the cauda equina in the remaining three. SEP recordings enabled us to clearly differentiate the response generated by the dorsal horns of the lumbosacral cord from the activity generated in the cauda equina dorsal roots. Neurophysiologic findings strongly correlated with the lesion site demonstrated by the MRI (cauda equina, lumbosacral cord, or thoracic cord immediately above the lumbar enlargement). The N24 cord potential was absent with a normal cauda equina response in 10 of 12 patients with MRI signs of lumbosacral cord lesions. Clinical examination often failed to localize the spinal cord as the lesion site, suggesting pure peripheral involvement in six of the 12 patients with MRI signs of lumbosacral cord lesions. PMID- 8232942 TI - Analysis of the c-FOS gene on chromosome 14 and the promoter of the amyloid precursor protein gene in familial Alzheimer's disease. AB - The c-FOS gene product, a putative transacting transcriptional regulator of the amyloid precursor protein (APP) gene, is a candidate locus for the familial Alzheimer's disease (FAD) mutation on chromosome 14 (FAD14). In light of this functional relationship, we investigated the nucleotide sequence and segregation of c-FOS and the nucleotide sequence of the 5' APP promoter. Single-stranded conformational polymorphisms (SSCPs) in the c-FOS gene revealed that c-FOS closely cosegregates with the FAD14 gene but does not show allelic association with FAD. A conservative third-position T-->C mutation was demonstrated in exon 2 (codon 84) of c-FOS, and a C-->G substitution was detected at -209 bp in the 5' promoter of APP. Neither were unique to FAD and are unlikely to be pathogenic or secondary modifiers of the FAD phenotype. We conclude that the c-FOS open reading frame is probably not the site of the FAD14 locus, but we cannot exclude the existence of modifier loci on chromosome 21. PMID- 8232943 TI - Carbamazepine withdrawal: effects of taper rate on seizure frequency. AB - We prospectively investigated the effects of rate of carbamazepine (CBZ) withdrawal and CBZ level on seizure type and frequency in 12 epilepsy patients withdrawn completely from antiepileptic drugs prior to entering an investigational monotherapy trial. Patients withdrawn from CBZ rapidly (over 4 days) experienced significantly more generalized tonic-clonic seizures (GTCSs) and GTCS clusters than did those withdrawn slowly (over 10 days). Complex partial seizure (CPS) frequency did not differ between the two groups. CPSs preceded GTCSs, with GTCSs occurring in the majority of patients after CBZ had been discontinued, at subtherapeutic or absent CBZ levels. Two of six patients who had been tapered rapidly and all six patients who had been tapered slowly were able to enter the investigational monotherapy trial. PMID- 8232944 TI - Placebo-controlled study of the efficacy and safety of lamotrigine in patients with partial seizures. U.S. Lamotrigine Protocol 0.5 Clinical Trial Group. AB - We evaluated the efficacy and safety of lamotrigine (300 and 500 mg/day) as add on therapy in a multicenter, randomized, double-blind, parallel-group, placebo controlled study of 216 patients with refractory partial seizures. During 6 months of treatment, median seizure frequency decreased by 8% with placebo, 20% with 300 mg lamotrigine, and 36% with 500 mg lamotrigine. Seizure frequency decreased by > or = 50% in one-third of the 500-mg group and one-fifth of the 300 mg group. Reductions in seizure frequency and seizure days were statistically significant, compared with placebo, for the 500-mg group but not the 300-mg group. Most adverse events were minor and resolved over time. Nine percent of patients on lamotrigine withdrew because of adverse experiences. Lamotrigine plasma concentrations appeared to be a linear function of dose, and the drug did not affect plasma concentrations of concomitant antiepileptic drugs. Lamotrigine was safe, effective, and well tolerated as add-on therapy for refractory partial seizures. PMID- 8232945 TI - Gabapentin as add-on therapy in refractory partial epilepsy: a double-blind, placebo-controlled, parallel-group study. The US Gabapentin Study Group No. 5. AB - Gabapentin, administered as add-on therapy, was safe and effective in this 12 week, multicenter, placebo-controlled, parallel-group study in 306 patients with refractory partial epilepsy. For patients in each gabapentin treatment group (600, 1,200, or 1,800 mg/d), the mean response ratio was significantly better than that of a placebo group. The percentage of patients achieving at least a 50% reduction in seizure frequency was 8% among placebo-treated patients and ranged from 18% to 26% for patients who received gabapentin. Adverse events were generally mild and transient and occurred at a slightly higher frequency among patients receiving gabapentin than among those receiving placebo. Gabapentin did not affect the serum concentrations of concurrent antiepileptic drugs and was not regularly associated with any deviations in clinical laboratory values. Gabapentin's low inherent toxicity and its lack of drug interactions make it an ideal candidate for use as add-on therapy in patients with refractory partial epilepsy. PMID- 8232946 TI - Comprehensive study of diagnosis and treatment of trigeminal neuralgia secondary to tumors. AB - Among 5,058 patients seen at the Mayo Clinic from 1976 through 1990 for face pain, we diagnosed trigeminal neuralgia in 2,972. Tumors were causing the face pain in 296 patients. Sex and pain distributions paralleled those in idiopathic trigeminal neuralgia; however, patients with tumors causing trigeminal neuralgia were younger than those with idiopathic pain. Meningiomas and posterior fossa tumors were the most common. Neurologic deficits developed on follow-up evaluation in 47% of the patients, often precipitating further study and eventual diagnosis of the tumor. Delay in tumor diagnosis averaged 6.3 years. CT with contrast was the most frequently used initial diagnostic radiographic technique, detecting a tumor in 40 of 43 examinations. MRI was subsequently used to confirm and better delineate the tumor in five of five cases. Carbamazepine was the most effective drug for relieving trigeminal neuralgia, but relief was usually temporary. Of the surgical treatment options, total removal of the tumor was the most effective in completely relieving tic pain. In patients at high surgical risk, however, temporarily or permanently blocking afferent impulses with radiofrequency ablation, glycerol rhizotomy, or alcohol blocks was a good alternative to craniotomy. PMID- 8232947 TI - Posthypoxic treatment with felbamate is neuroprotective in a rat model of hypoxia ischemia. AB - Felbamate, a novel dicarbamate anticonvulsant that blocks the glycine site of the N-methyl-D-aspartate receptor and protects the hippocampal slice from hypoxic damage, shows remarkably low toxicity in animals and in humans. Since most treatment of human cerebral ischemia will have to be delivered after the insult, we investigated the neuroprotective potency of post hoc felbamate in rat pups with bilateral carotid ligations exposed to an atmosphere of 6.5% O2 for 1 hour. Brain temperature was unaffected by surgery, hypoxia, or felbamate. Neuroprotection was greatest at 300 mg/kg, less effective at 200 and 400 mg/kg, and ineffective at 100 mg/kg. Post hoc felbamate (300 mg/kg) reduced the volume of infarction from 67% +/- 7% of neocortex in unmedicated rats to 32% +/- 8%, 51% +/- 12%, 38% +/- 19%, and 53% +/- 10% when given 0, 1, 2, and 4 hours after hypoxic exposure, respectively. By 6 hours, post hoc protection was no longer significant. Delayed neuronal necrosis in hippocampal granule cells was reduced from 156 +/- 33 neurons to 12 +/- 7 (0 hours, p < 0.01) and 37 +/- 17 (1 hour, p < 0.05). These effects were obtained at plasma concentrations (60 to 120 mg/ml) that have occasionally been reached without serious toxicity in human anticonvulsant trials. These data suggest that, in this animal model, felbamate given after a hypoxic-ischemic insult is effective in reducing cerebral infarction and extremely effective in preventing delayed neuronal necrosis, but that the window of opportunity for post hoc treatment is only 1 to 4 hours. PMID- 8232948 TI - Functional magnetic resonance imaging of complex human movements. AB - Functional magnetic resonance imaging (FMRI) is a new, noninvasive imaging tool thought to measure changes related to regional cerebral blood flow (rCBF). Previous FMRI studies have demonstrated functional changes within the primary cerebral cortex in response to simple activation tasks, but it is unknown whether FMRI can also detect changes within the nonprimary cortex in response to complex mental activities. We therefore scanned six right-handed healthy subjects while they performed self-paced simple and complex finger movements with the right and left hands. Some subjects also performed the tasks at a fixed rate (2 Hz) or imagined performing the complex task. Functional changes occurred (1) in the contralateral primary motor cortex during simple, self-paced movements; (2) in the contralateral (and occasionally ipsilateral) primary motor cortex, the supplementary motor area (SMA), the premotor cortex of both hemispheres, and the contralateral somatosensory cortex during complex, self-paced movements; (3) with less intensity during paced movements, presumably due to the slower movement rates associated with the paced (relative to self-paced) condition; and (4) in the SMA and, to a lesser degree, the premotor cortex during imagined complex movements. These preliminary results are consistent with hierarchical models of voluntary motor control. PMID- 8232949 TI - Bromocriptine-induced dystonia in patients with aphasia and hemiparesis. AB - Five of seven patients with chronic nonfluent aphasia and hemiparesis due to a focal ischemic infarction developed painful hemidystonia during treatment with a high dose of bromocriptine. All seven patients had cortical damage, but four also had basal ganglia and one thalamic involvement. While lesion location did not differ between dystonic and nondystonic patients, the dystonic patients had more weakness than those without dystonia. PMID- 8232950 TI - Infantile spasms: outcome and prognostic factors of cryptogenic and symptomatic groups. AB - We reviewed the outcome (developmental, neurologic, and seizure) and prognostic factors of 57 cases of infantile spasms (17 cryptogenic, 40 symptomatic). The mean developmental score of the cryptogenic group (71.2 +/- 24.2) was significantly higher than that of the symptomatic group (48.4 +/- 24.5), as assessed by the Griffith Mental Developmental Scale. A resultant neurologic deficit was present in 23.5% of the cryptogenic group and 75.0% of the symptomatic group. Coexistence of other forms of seizures was observed in 35.3% of the cryptogenic group and 57.5% of the symptomatic group. Outcome was affected by the coexistence of other seizures, presence of neurologic deficit, time lag in initiation of treatment (cognitive outcome only), poor response to ACTH treatment, and persistent EEG abnormality. Character of spasms and time lag in initiation of treatment (seizure outcome only) did not have any significant effect on outcome. PMID- 8232951 TI - Contrast sensitivity dysfunction in Alzheimer's disease. AB - We compared the spatial contrast sensitivity of six patients with mild to moderate Alzheimer's disease (AD) and six age-matched control subjects in a parametric design. Results demonstrate reduced contrast sensitivity in patients with AD at all but the lowest frequency tested. The results suggest that the effect of AD on spatial contrast sensitivity is stronger at higher frequencies and provide a rationale for complaints of poor vision in AD patients. PMID- 8232952 TI - Scrapie prions alter receptor-mediated calcium responses in cultured cells. AB - The molecular basis of neurologic dysfunction in prion diseases is unknown. Spongiform degeneration of neurons is the most characteristic neuropathologic change which raises the possibility of abnormal ion channel function. Here we examined the regulation of Ca2+ fluxes in two cell lines chronically infected with scrapie prions, designated ScN2a (scrapie-infected mouse neuroblatoma) and ScHaB (scrapie-infected hamster brain) cells. In uninfected HaB cells, bradykinin caused increases in intracellular Ca2+ concentration ([Ca2+]i) by release of Ca2+ from internal stores and influx of extracellular Ca2+ whereas, in N2a cells, bradykinin increased [Ca2+]i exclusively from internal stores. Prion infection of both cell lines markedly reduced or eliminated bradykinin-activated increases in [Ca2+]i, whether driven by internal or extracellular sources. Stressing the cells with high extracellular [Ca2+], 8 to 20 mM, led to cytopathologic changes in ScHaB but not in ScN2a cells. Cytopathology was not preceded by an increase in [Ca2+]i. These findings indicate that scrapie infection induces abnormalities in receptor-mediated Ca2+ responses and raise the possibility that nerve cell dysfunction and degeneration in prion diseases is related to ion channel aberrations. PMID- 8232953 TI - Cardiomyopathy may be the only clinical manifestation in female carriers of Duchenne muscular dystrophy. AB - Cardiomyopathy was reported in a few Duchenne muscular dystrophy (DMD) carriers with clinical evidence of myopathy. We report two carriers with dilated cardiomyopathy, increased serum CK, and no symptoms of muscle weakness. In heart biopsies of both patients, dystrophin-the protein product of DMD locus--was absent in many fibers. Dilated cardiomyopathy may be the only manifestation of dystrophin gene mutation in carriers. PMID- 8232954 TI - Cortical tremor. A common manifestation of cortical myoclonus. AB - Ten patients, three with postural tremor and seven with action myoclonus, had stereotyped involuntary rhythmic movements when attempting to execute a sustained isometric muscle contraction. The movements were characterized by rhythmic EMG bursts lasting less than 50 msec and appearing synchronously in agonist and antagonist muscles at a rate of 9 to 18 Hz. Backaveraging of the EEG activity related to the onset of the rhythmic EMG bursts identified a cortical potential preceding the EMG bursts in all patients. These symptoms and signs fit the description of "cortical tremor," a variant of cortical reflex myoclonus. Cortical tremor is common in patients with cortical myoclonus and may be a source of functional disability. In two patients in whom we studied the effects of graded levels of isometric force, force recruitment modulated the abnormal EMG bursting frequency, amplitude, and spatial distribution of the myoclonic jerks in the activated limb. Transcranial magnetic and electrical stimulation, but not peripheral nerve stimulation, influenced the abnormal EMG bursting pattern, implying a greater dependence of this rhythmic phenomenon on a central generator than on peripheral feedback loops. PMID- 8232955 TI - Multiple dimensions of handedness and the human corpus callosum. AB - Consistency of right-hand preference correlates with callosal isthmus size. Anterior corpus callosum (CC) regions vary as a function of sex and right- versus left-hand preference. We analyzed regional CC anatomy in a sample of right- and left-handed men and women on the basis of both degree ("Consistency") and direction ("Hand") of hand preference scores. Significant Hand x Consistency effects indicated that both are important factors in CC organization. These data also suggested that a single continuum of hand preference and regional CC size may not be applicable across all subjects. PMID- 8232956 TI - Expression of Epstein-Barr virus proteins in primary CNS lymphoma in AIDS patients. AB - We examined the expression of the Epstein-Barr virus (EBV)-induced proteins (LMP [latent membrane protein], EBNA-2, and CD23) and a lytic protein, viral capsid antigen (VCA), in five acquired immune deficiency syndrome (AIDS)-related primary CNS lymphomas (PCNSLs). We compared that expression with the expression of the same proteins in PCNSL from six immunocompetent patients and severe combined immune deficiency (SCID) mouse brains injected with EBV-infected lymphoblastoid cell lines (LCLs). Brain biopsy tissue from an AIDS patient with progressive multifocal leukoencephalopathy (PML) and a normal brain was also studied. Three of the AIDS PCNSLs expressed both human immunoglobulin kappa and lambda light chains and two expressed lambda light chain only. All non-AIDS-related PCNSLs expressed a single light-chain isotype. All five AIDS-related PCNSLs expressed LMP-1 (> 40%), EBNA-2 (> 60%), and VCA (1 to 5%) of tumor cells. These proteins were similarly expressed in the SCID/human chimeras. None of the PCNSLs from immunocompetent subjects, the normal brain, or the brain of the patient with PML expressed these proteins. PCNSL in AIDS patients bears greater similarity to EBV infected LCLs than to PCNSL from immunocompetent patients. PMID- 8232957 TI - Impaired fatty acid oxidation in children on valproic acid and the effect of L carnitine. AB - Fatty acid oxidation was studied in 12 patients (aged 3 to 19 years) receiving valproic acid (VPA), predominantly as monotherapy, before and after 1 month of L carnitine supplementation (50 mg/kg/day po) in order to determine whether L carnitine plays a role in preventing the hepatotoxic effects of this drug. Five of these patients were also studied prior to VPA treatment. Only one patient taking VPA had an abnormally low plasma free carnitine. Acyl-/free carnitine ratios were elevated in five patients on VPA and normalized after L-carnitine supplementation. Mean plasma concentrations of free fatty acids, beta-OH butyrate, and cumulative excretion of 13CO2 after administration of 1-13C octanoic acid were not changed by VPA or L-carnitine treatment. Urinary dicarboxylic acids, acylglycines, and octanoylcarnitine were elevated during VPA therapy and unaltered by L-carnitine. These results suggest that, in patients at low risk for VPA-induced hepatotoxicity (patients aged > 2 years and taking VPA as monotherapy), VPA causes metabolic abnormalities resembling those found in inborn errors of mitochondrial beta-oxidation which are not corrected by L carnitine. PMID- 8232958 TI - Genetics of facioscapulohumeral muscular dystrophy: new mutations in sporadic cases. AB - A gene for facioscapulohumeral muscular dystrophy (FSHD) has been linked to chromosome 4q35 in families with the disease. We have used recently characterized p13E-11/D4S809 probes that map near or within the FSHD gene to investigate eight sporadic cases of FSHD whose parents showed no signs of disease. Probe p13E 11/D4S809 detected novel DNA fragments in seven of the eight sporadic FSHD individuals and not in the parents, substantiating the clinical diagnosis. Two sisters with FSHD whose parents were clinically normal had a novel DNA fragment suggestive of germline mosaicism. Probe p13E-11/D4S809 is potentially helpful in genetic counseling. However, because this probe may also detect a locus unlinked to chromosome 4, because of possible genetic heterogeneity in FSHD, and because of the presence of recombinants in autosomal dominantly inherited families, closer markers or gene definition will be needed for accurate genetic counseling in other situations. PMID- 8232959 TI - Intranuclear rods in severe congenital nemaline myopathy. AB - We compared the muscle pathology and clinical course in eight patients with congenital nemaline myopathy. An abundance of large intranuclear rods was present in the muscle fibers of one patient with a rapid, fatal course. Intranuclear rods were not present in the muscles of seven patients with a benign course. The large intranuclear rods and the smaller sarcoplasmic rods were similar ultrastructurally and exhibited positive immunoperoxidase staining with anti alpha-actinin antibodies. The accumulation of alpha-actinin within myonuclei may reflect a severe disturbance of normal intracellular processes regulating myofibrillar synthesis. Since two previously reported infants with intranuclear nemaline rods also had a fatal outcome, the presence of intranuclear rods may represent a marker for a severe form of congenital nemaline myopathy. PMID- 8232960 TI - Conjugal amyotrophic lateral sclerosis: report of a young married couple. AB - We report a 38-year-old nurse who developed amyotrophic lateral sclerosis (ALS) beginning in September 1990. In May 1991, her 38-year-old husband developed dysarthria, which progressed to typical ALS. This is the fourth report in the literature of conjugal ALS occurring outside of Guam. Although this event is most likely due to coincidence, exogenous agents should be considered in the etiology of ALS. PMID- 8232961 TI - MRI assessments of hippocampal pathology in extratemporal lesional epilepsy. AB - We performed a prospective study in 18 patients to determine the extent of MRI identified hippocampal pathology in patients with intractable partial epilepsy of extratemporal origin. A mesial temporal signal-intensity alteration or hippocampal formation (HF) atrophy, or both, have been shown to be reliable markers of the temporal lobe of seizure origin in patients with mesial temporal sclerosis. All patients subsequently received surgical ablative therapy between 1988 and 1992. During shortterm follow-up, 14 of the 18 patients experienced a significant reduction in seizure tendency, and 12 patients were rendered seizure free. Qualitative and quantitative (HF volumetry) assessments of HF pathology were performed retrospectively by a blinded investigator. No hippocampal imaging alteration was present in 17 patients. Left HF atrophy was confirmed in one patient with post-traumatic epilepsy who underwent a successful right frontal lobectomy. Morphometric MRI studies rarely identify hippocampal pathology in patients with extratemporal epilepsy. PMID- 8232962 TI - Essential tremor and dystonia. AB - We reviewed the database of the Dystonia Clinic at the University of Kansas Medical Center for patients with dystonia and tremor. Of 296 patients with idiopathic dystonia, 24 had dystonic tremor, 20 with cervical dystonia had an isolated head-nodding tremor, two patients with writer's cramp had ipsilateral hand tremor, and two patients with generalized dystonia had arm tremor. Eight patients, all with cervical dystonia, had essential tremor that preceded the onset of their dystonia. PMID- 8232963 TI - Progressive myoclonus epilepsy of Unverricht-Lundborg type: a clinical and molecular genetic study of a family from the United States with four affected sibs. AB - We describe clinical and molecular genetic data on a family from the United States in which four of five sibs are affected with progressive myoclonus epilepsy of Unverricht-Lundborg type. The gene for this disorder (EPM1) has previously been mapped to the distal region of chromosome 21. Molecular genetic results suggest that the disease gene in this family is linked to the same region of chromosome 21. Crossover events in the family help refine the gene localization by placing EPM1 between loci CBS and D21S112. PMID- 8232964 TI - Choroido-cerebral calcification syndrome with retardation. AB - We present a sibship with a rare syndrome characterized by mental retardation, dense calcification of the lateral ventricular choroid plexus, and increased CSF protein. Neurophysiologic studies yielded nonspecific results, and endocrine studies, including parathormone levels, were normal. Simultaneous measurements of CSF and serum calcium, magnesium, and other electrolytes were normal, but the CSF/serum ratio of phosphate was low, suggesting a possible role in the pathogenesis of this syndrome. PMID- 8232965 TI - Complex partial status epilepticus in a patient with dural metastases. AB - We describe a 68-year-old man with invasive transitional cell carcinoma of the bladder metastatic to the dura who presented with complex partial status epilepticus (CPSE). To our knowledge, the association of CPSE and dural metastases has not been previously reported. PMID- 8232966 TI - A new (two-repeat) octapeptide coding insert mutation in Creutzfeldt-Jakob disease. AB - We report a family in which the proband died of clinically typical, neuropathologically verified Creutzfeldt-Jakob disease; her still-living mother suffers from a progressive dementia of many years' duration, and her maternal grandfather died after a similar illness. The proband, her mother, and two of three young first-degree relatives all have an identical insert mutation in the PRNP gene consisting of a twice-repeated 24-nucleotide sequence in the region between codons 51 and 91. PMID- 8232967 TI - Interventional neurology: a subspecialty whose time has come. PMID- 8232968 TI - Practice parameter for carpal tunnel syndrome (summary statement). Report of the Quality Standards Subcommittee of the American Academy of Neurology. PMID- 8232969 TI - Sensory neuronopathy in autoimmune chronic active hepatitis. PMID- 8232970 TI - Musculocutaneous mononeuropathy complicating Capnocytophaga canimorsus infection. PMID- 8232971 TI - Stiff-man syndrome treated with intrathecal baclofen. PMID- 8232972 TI - The Clinical Dementia Rating (CDR): current version and scoring rules. PMID- 8232973 TI - 'Reappearance' of congenital nystagmus after minor head trauma. PMID- 8232974 TI - Reversible paraneoplastic limbic encephalitis. PMID- 8232975 TI - Mesencephalic ependymal cyst causing an inferior divisional paresis of the oculomotor nerve: case report. PMID- 8232977 TI - CJD in health care workers. PMID- 8232976 TI - Cytomegalovirus radiculomyelitis. PMID- 8232978 TI - Somesthetic syndrome. PMID- 8232979 TI - Kennedy's disease. PMID- 8232980 TI - MRI in ataxias. PMID- 8232981 TI - HLA and MS. PMID- 8232982 TI - Emerging antiepileptic drugs. AB - The introduction of several new antiepileptic drugs in the United States is likely in 1993. Many new drugs have undergone testing, but the four currently considered the most important are felbamate, gabapentin, lamotrigine, and vigabatrin. When these drugs are used as add-on therapy for patients with intractable epilepsy, 20 to 60% of patients show at least a 50% improvement in seizure frequency and 7% become seizure free. An overview of these agents is presented. PMID- 8232983 TI - The role of vigabatrin in the management of infantile epileptic syndromes. AB - More than 360 children with intractable epilepsy have been treated with vigabatrin in single-blind or open, add-on studies. Approximately 50% or more of patients with West syndrome and partial seizures have shown a 50% or greater reduction in seizure frequency with the use of vigabatrin. A less consistent response has been found between studies evaluating vigabatrin in children with Lennox-Gastaut syndrome, although, overall, approximately 50% of these patients have also shown a greater than 50% decrease in seizures. The use of vigabatrin in idiopathic localization-related epilepsy, idiopathic generalized epilepsy, and the Landau Kleffner syndrome have not been reported, but its evaluation in these conditions may be warranted based on the relatively excellent safety profile of vigabatrin. Vigabatrin has been shown to aggravate "nonprogressive myoclonic epilepsies." Vigabatrin has been well tolerated in children, with mild drowsiness and agitation being the most commonly reported side effects. PMID- 8232984 TI - The potential for increasing seizure frequency, relapse, and appearance of new seizure types with vigabatrin. AB - Although generally effective, various antiepileptic drugs have been reported on occasion to increase seizure frequency, result in seizure relapse, or elicit new types of seizures. Some seizure types and epilepsy syndromes appear more prone than others to exacerbation by a given drug. Vigabatrin is rarely associated with seizure worsening, and then mainly in patients with resistant generalized epilepsies. PMID- 8232985 TI - Surgery for intractable seizures in infancy and early childhood. AB - Epilepsy surgery is becoming an increasingly used therapy for infants and young children with severe, medically intractable seizures. As in older children and adults, the presurgical evaluations of possible candidates typically consist of a detailed history, neurologic and neuropsychologic examination, and anatomic and functional neuroimaging. The "gold standard" test, however, is the recording of ictal events using simultaneous EEG and video monitoring. While temporal lobe resection is the most commonly performed surgery in older children and adults, nontemporal lobe resections, corpus callosotomies, and hemispherectomies are more commonly performed in younger children. Antiepileptic drugs remain the mainstay of treatment of children with epilepsy. However, the clinician should consider surgery early in the course of the catastrophic seizure disorders of childhood: infantile spasms, Sturge-Weber syndrome, and Rasmussen's encephalitis. PMID- 8232986 TI - Seizures in the developing brain. AB - Clinical and animal data show that there is a period of increased susceptibility to seizures during early postnatal development. The cause of this increased susceptibility is probably a combination of enhanced excitation and diminished inhibition throughout the brain, as well as developmental differences in subcortical circuits, such as the substantia nigra-mediated seizure-suppression system. Animal studies on the effect of prolonged epileptic seizures early in development indicate that even severe seizures produce little or no damage in the rat pup hippocampus. Several studies have demonstrated both qualitative and quantitative age-dependent differences in the effect of antiepileptic treatment. More studies are needed to determine the effects of long-term antiepileptic treatment. PMID- 8232987 TI - Classifying epileptic syndromes: problems and a neurobiologic solution. AB - Because the effectiveness of an antiepileptic drug in controlling a particular seizure type and the prognosis for remission may differ between epileptic syndromes, accurate syndrome classification is crucial. However, a lack of universally accepted diagnostic criteria for some syndromes makes precise classification impossible in many patients. When syndromic classification is not possible, it is often more useful to consider genetic, clinical, electroencephalographic, neuropsychologic, radiologic, and other laboratory factors in characterizing the epileptic condition and selecting the optimal treatment. PMID- 8232988 TI - Assistance for reservists called to active duty. PMID- 8232989 TI - Foodborne gastroenteritis. PMID- 8232990 TI - Another viewpoint. PMID- 8232991 TI - Automated external defibrillator training and skills maintenance in Air Force emergency medical services systems. AB - In April 1989, we initiated a test program to evaluate the use of semi-automated external defibrillators in five United States Air Force Medical Treatment Facilities. The objectives of the study were 2-fold. First, the feasibility of conducting a uniform training program in a number of geographically separated basic life support emergency medical services (EMS) systems was assessed. The second objective was evaluation of psychomotor skills maintenance by Air Force Emergency Department emergency medical technicians (EMTs) in simulated resuscitations 6 months after initial training. Performance was evaluated in each of six areas for the 48 EMTs completing retesting. Forty-three of the 48 EMTs were assessed to have properly completed the resuscitations. Three of the remaining five failed to clear prior to defibrillating, and in two cases the delivery of three defibrillatory shocks was delayed for 6-8 minutes from the time of arrival at the patient's side. These results demonstrate the feasibility of providing training and maintaining proficiency in military EMS systems. PMID- 8232992 TI - Dental epidemiology of military operations. AB - In this paper we review studies that have been conducted on the epidemiology of oral, dental, and maxillofacial conditions during military deployments. The limitations of our current knowledge base are discussed, as is a proposed research effort to enhance the responsiveness of dental support in theaters of operation. PMID- 8232993 TI - Death investigations on military installations. AB - The authority to conduct a death investigation, including an autopsy examination, on a military installation depends upon the type of jurisdiction on the installation and the decedent's military connection. Exclusive federal jurisdiction gives the military authority to investigate deaths of active duty military members and, under specific conditions, of civilians with a military connection. Concurrent or shared jurisdiction gives the local civilian medical examiner the right of first refusal as to military members. Thus, the civilian medical examiner may waive jurisdiction to the military. This is not the case when a civilian dies under the same circumstances. PMID- 8232994 TI - Fish and other aquatic animal kills: causes and relevance to public health for military health care professionals. AB - Environmental contamination is a public health issue. Fish or other aquatic animal kills can be a sign that contamination of the aquatic environment has occurred. They are highly visible to the public. When such kills occur in water bodies on military installations, medical personnel are often involved in the investigation to assess the public health implications of the kill or its causes. This article discusses the causes of aquatic animal kills, describes a proper response to a kill, and gives several case studies. PMID- 8232995 TI - Discharge planning and diagnosis-related groups in military hospitals: key issues and essential elements. AB - As pressures for cost containment in health care continue to mount, military hospitals will inevitably be affected. Cost-containment strategies used in civilian hospitals such as diagnosis-related groups will continue to be implemented in military health care facilities. Effective discharge planning is essential for dealing successfully with pressures for cost containment. Key elements in an effective discharge planning program are discussed. PMID- 8232996 TI - Explaining health-promoting lifestyles of Navy personnel. AB - This study examined the relationship of six dimensions of health-promoting lifestyle (self-actualization, health responsibility, interpersonal support, exercise, nutrition, and stress management) with selected sociodemographic and attitudinal variables in a random sample of 421 active duty Navy personnel. Participants who were older, officers, shore-based, married, and who reported high levels of perceived health all had significantly higher scores for overall health-promoting life-style and several lifestyle dimensions. Exercise of self care agency--a composite health attitude indicator--accounted for 35% of the variance in health-promoting lifestyle patterns. Among the sociodemographic variables, educational level, race, and perceived health state were differentially predictive of healthy lifestyle dimensions. PMID- 8232997 TI - Changing presentation of coronary heart disease in an inpatient population within the U.S. military health care system. AB - This study examines the changing presentation of coronary heart disease (CHD) in an inpatient population at Brooke Army Medical Center. The specific objectives of the study were to examine the presentation of CHD in a population unbiased by diagnosis-related group (DRG) reimbursements and to assess the importance of unstable angina and prior history of disease in the presentation of CHD. One thousand fifteen discharges in 1985 and 1,304 discharges in 1990 with the diagnosis of CHD were reviewed by cardiologists for evidence of symptomatic heart disease at the time of hospitalization. Forty percent of these charts were accepted into this study. The presentation rates of CHD were 1% with sudden death, 26% with myocardial infarction, 64% with angina, and 9% with congestive heart failure (CHF). During the study period, stable angina, Q-wave infarctions, and the myocardial infarction case fatality rate decreased (p < 0.05) and CHF and non-Q wave infarcts increased (p < 0.001). However, unstable angina was the most common presentation of CHD, and differences (p < 0.05) were noted in the presentation of CHD in patients with and without a prior history of disease. This study demonstrates the significance of unstable angina and prior history of CHD in an environment free of bias from DRG reimbursements. PMID- 8232998 TI - An evaluation of the U.S. Air Force's detailed follow-up audiometric examination program. AB - The authors used information on individuals demonstrating permanent threshold shifts contained in the United States Air Force Hearing Conservation Data Registry to evaluate the effectiveness of detailed follow-up audiometric examinations as part of the USAF Hearing Conservation Program. Analysis yielded a relative risk of 1.00 with a 95% confidence interval of 0.97-1.03, which indicates a high probability that the true effect of these audiograms on the results of the next annual exam is minuscule, if not zero. PMID- 8232999 TI - Heat illness and soldiers. AB - Soldiers are probably more at risk for heat illness than marathon runners and the reasons for this contrast are examined. The epidemiology of heat illness is reviewed, the practice of making soldiers run in battle-order is criticized, and further areas for research are suggested. PMID- 8233000 TI - Psychological assessment for security clearances, special access, and sensitive positions. AB - This paper describes issues in the psychological assessment of individuals for security clearances, special duty, and sensitive positions. Using assessment for access to sensitive compartmented information as an example, the paper identifies, and explicates problems in criterion validity (the NEXUS issue), practical approaches to resolving these problems, selection of assessment techniques, and writing of the assessment report. PMID- 8233001 TI - Survey of the Navy's three-tiered obesity treatment program. AB - The Navy's diverse, three-tiered obesity treatment program is described. Level I (command-directed) programs rely primarily on group exercise to treat obesity; most level II (outpatient counseling) and level III (6-week inpatient) programs are modeled on Overeaters Anonymous and devote substantial amounts of time to group discussion, behavior modification, and nutrition education. Lack of funding or staffing has prevented many level II facilities from conducting a weight management program, however. Further research might explore the potential for level II to provide a cost-effective middle ground for obesity treatment. PMID- 8233002 TI - Prevalence of asymptomatic chlamydial cervical infection in active duty Army females. AB - Four hundred seventy-six consecutive active duty Army females who presented for routine pap smears were screened for Chlamydia trachomatis and Neisseria gonorrhea. Thirty-nine of 476 (8.2%) tested positive for chlamydia using the Chlamydiazyme enzyme immunoassay. All patients with positive tests for chlamydia were asymptomatic and had normal pelvic exams. The average age of patients testing positive for chlamydia was 23.9. Only 6 of the 39 patients with chlamydia were older than 30. Tests for gonorrhea and pap smear results had little correlation with patients testing positive for chlamydia. The high prevalence of chlamydia in this population of asymptomatic women makes it probable that screening similar populations of patients would be more cost-effective than treating the complications of this disease. PMID- 8233003 TI - Acute myocardial infarction mortality in VA versus non-VA settings. AB - Mortality associated with acute myocardial infarction has declined steadily over the past 25 years. This study was conducted to compare in-hospital mortality among men treated in VA versus non-VA settings. Results supported declines in mortality since 1980 to an overall rate of 11%, as reported in published literature and found in the VA data base. Mortality among men treated in VA settings mirrored rates reported for non-VA settings, both overall and by age groups. Findings were shared with VA Medical Centers for use in identifying opportunities to improve clinical practice. PMID- 8233004 TI - Advanced trauma life support: assessment of cognitive achievement. AB - One hundred fifteen physicians participating in advanced trauma life support (ATLS) training courses had entry (pre-test) and exit (post-test) level knowledge compared. There was significant improvement for the overall group mean score from the pre-test to post-test (p = 0.0001). Comparisons revealed significant differences for pre-test and post-test scores by practice specialty and level of training. Residents in training had significantly higher scores for both pre-test (p = 0.0177) and post-test (p = 0.0024) than did the practicing physician group. No significant differences were detected in the cognitive achievement between the residents according to practice specialty. PMID- 8233005 TI - The implementation of the composite health care system into total quality management in military medical treatment facilities. AB - The Composite Health Care System (CHCS) is a new medical computer system which will integrate into Total Quality Management (TQM) in Department of Defense hospitals. Both are examples of integrated systems which mandate a complete reshaping of the way the business of patient care is conducted. As a medical information management system, CHCS is an effective component of TQM and follows W. Edward Deming's TQM principles. With proper training, patience, and full support from end user and senior staff, CHCS and TQM can work together to maintain high quality standards for patient care in today's rapidly changing military medical service. PMID- 8233006 TI - Ultrasonographic appearance of malignant fibrous histiocytoma of the spermatic cord: a case report. AB - A case of malignant fibrous histiocytoma of the spermatic cord with ultrasound features is presented. The tumor rarely affects the spermatic cord. Our case is only the second reported in the literature with ultrasound correlation; in addition, our sonographic findings differ from the first reported case and more closely approximate the sonographic appearance of this tumor elsewhere in the body. PMID- 8233007 TI - Granulomatous Pneumocystis carinii in AIDS patients. AB - Seven cases of Pneumocystis carinii pneumonia with granulomatous reaction in patients infected with the human immunodeficiency virus are described. The patients were all adult men between the ages of 32 and 45 years, with different high-risk factors. Clinically, all the patients presented with a history of non productive cough and shortness of breath. Two of the patients had a past history of pulmonary pneumocystosis. Radiologically, six patients had diffuse pulmonary infiltrates and one nodular pulmonary infiltrate. Transbronchial lung biopsies were obtained in four patients and open lung biopsies in three. All presented a predominant granulomatous reaction composed of epithelioid and multinucleated giant cells. Several other special stains to detect the presence of other microorganisms to account for the granulomatous reaction were negative. PMID- 8233008 TI - [The viable myocardium: metabolic, diagnostic and therapeutic aspects]. AB - The term stunned myocardium is used to indicate a reversible post-ischemic dysfunction of the ventricular mechanism which may persist for hours, days or weeks after the restoration of coronary flow following spontaneous or pharmacological thrombolysis, transluminal coronary angioplasty, aorto-coronary bypass and ischemic attacks. Hibernating myocardium is used to describe a depression of ventricular contractility in the presence of chronic hypoperfusion which may be reversed following revascularization as a result of aorto-coronary by-pass surgery. Three biochemical and physiopathological hypotheses are currently acknowledged to explain the phenomenon of stunning: the hypothesis of free oxygen radicals, the hypothesis related to an energy deficit and that involving a calcium overload. It is possible that oxydizing stress induced by free radicals may modify the activity of one or more sarcolemmic proteins which regulate the flow of calcium or other ions. Alterations in the transport and accumulation of calcium ions due to a Na+/Ca++ pump deficit and calcium-ATPase of the sarcoplasmatic reticle appear to be responsible for contractile dysfunction. The hypothesis concerning an energy deficit appears to be least probable since even if ATP levels are low the intracellular energy status does not appear to be a factor which limits mechanical function which may be stimulated in the absence of further variations in the content of highly energetic phosphates. There is also reduced myofibrillar creatinkinase activity. In hibernating myocardium the mechanical dysfunction is due to a metabolic and therefore contractile "down regulation' with low myocardial energy and oxygen consumption to ensure the survival of chronically hypoperfused areas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233009 TI - [Carotid kinking and arterial hypertension. Preliminary results of the OPI program]. AB - The relationship between carotid kinking and arterial hypertension is stressed, based on the results of the first two years of an epidemiological research on prevalence of carotid lesions capable of producing cerebral ischemia in a population, aged 45 to 75, resident within the district USSL no. 69 (Parabiago), Lombardia. Reported data are referred to 13,936 subjects that underwent the first level of screening between January 29th 1990 and December 31st 1991. 1,386 subjects underwent further examination by color coded echo flow (second level of screening) due to suspected carotid lesion; they were divided into two groups according to the presence or absence of arterial hypertension. The distribution of these groups does not repeat that of the general population (p < 1 x 10(-6)). 158 subjects were affected with carotid kinking. Within the normotension group (439; 31.67%), 39 (8.88%) were demonstrated to harbor kinkings, while in the hypertension group (947; 68.33%) 141 (14.88%). Thus, the difference between the two groups remains largely below the limits of statistical significance (p > 1). The association between kinking and hypertension is not affected by sex (p > 1 x 10(-1)), regardless of a significant difference of distribution between the sexes (145 women and 40 men: p < 1 x 10(-6)). The conclusions point out that the evidence of this study strongly stresses that the association between carotid kinking and arterial hypertension is incidental. PMID- 8233010 TI - Left ventricular hypertrophy and diastolic dysfunction in alcohol-associated hypertension. AB - In order to assess left ventricular structure and diastolic function, 50 hypertensive male subjects, 25 with and 25 without a history of alcohol abuse, and 20 normotensive male controls underwent Doppler echocardiographic examination followed by ambulatory blood pressure monitoring for 24 hours. Left ventricular mass was significantly higher in alcoholic hypertensives in relation to non alcoholic hypertensives (p < 0.05) and normotensive controls (p < 0.001). Moreover, Doppler parameters expressing left ventricular filling pattern were significantly worse in alcoholic than in non-alcoholic hypertensives (p < 0.01). Clinic and ambulatory blood pressure were similar in alcoholic and non-alcoholic hypertensives, while mean day-time heart rate was significantly higher in alcoholics (p < 0.01). Collected data suggest that non-hemodynamic factors are probably involved in the development of cardiovascular abnormalities in hypertensive alcoholics, and that echocardiography should be employed for risk profile definition in alcohol-associated hypertension. PMID- 8233011 TI - [Arrhythmia in thalassemia major: evaluation of iron chelating therapy by dynamic ECG]. AB - Arrhythmias and sudden death represent striking features in the natural history of thalassemia major. Antiarrhythmic treatment, however, does not appear to change the clinical course. During recent years the disease's therapeutics approach has undergone a substantial evolution, being more adequate the transfusional regimens as well as more effective the iron chelation therapy through subcutaneous infusion of deferoxamine. The aim of the present study was to determine possible influences exerted by the current treatment upon disease's arrhythmic disorders. Thirty patients of both sexes were enrolled in the study. The age ranged from 9 to 24 years. No congenital or acquired heart diseases were present. Each patient underwent concentrated red cell transfusions (in order to obtain pretransfusional hemoglobin levels of 10-11 g%), and iron-binding therapy through continuous subcutaneous microinjection of deferoxamine 40-50 mg/kg/day (6 8 hours/day, 6 days/week). Patients were divided in 2 groups: the first group (group A) comprising the 16 patients with good therapeutic compliance and regular pharmacological regimen; the second group (Group B) including the remaining poorly compliant 14 patients. The following parameters were analyzed: age, average hemoglobin levels during the last year, total amount of red cell transfusions, ferritin levels, starting age of iron-binding therapy. Moreover, each patient underwent 24-hour ECG Holter monitoring. Age (Group A: 18 +/- 4.6; Group B: 14 +/- 2.7; p < 0.02), total amount of transfusions (Group A: 272 +/- 73; Group B: 211 +/- 44; p < 0.03), and ferritin levels (Group A: 1697 +/- 860; Group B: 2908 +/- 730; p < 0.002) proved to be significantly different in the two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233012 TI - [Risk factors for arteriosclerosis and diet of surgical patients. 10 years of outpatient activity]. AB - The follow-up of arteriopathic subjects who have already been hospitalized (abdominal-peripheral vascular district 79% of patient and supra-aortic branch district 21%) has been carried out for around 10 years using an out-patient regime at the Institute of General and Cardiovascular Surgery of Milan. During outpatient visit the main risk factors for arteriosclerosis are routinely checked and treated if required; among the various therapies for metabolic control particular emphasis is placed on diet since its influence on the metabolism is well known, above all in the long term. The efficacy of this treatment has been evaluated by evaluating blood chemical changes (total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, VLDL and glycemia) in a group of patients who followed the diet in comparison to a group which did not. The results obtained show that in dieting patients there was a statistically significant decrease in blood values for total cholesterol, LDL cholesterol, triglycerides and VLDL, together with a decrement, which failed to reach statistical significance, in glycemia and HDL cholesterol. In non-dieting patients it was found that all parameters increased but that this was only statistically significant for total cholesterol and HDL cholesterol. The control of risk factors for arteriosclerosis through diet therapy therefore appears to be satisfactory even for secondary prevention in surgical arteriopathic subjects. It is important to underline that these results were not obtained under conditions of clinical research but in the reality of day-to-day clinical and therapeutic activity whose efficacy is vitally important for scientific health. PMID- 8233013 TI - [Serial Doppler echocardiography follow-up studies in the postoperative evaluation of severe pulmonary hypertension following surgery for mitral and mitral-aortic defects]. AB - Between January 1987 and December 1991 26 patients with mitral and mitro-aortic disease and severe pulmonary hypertension (> or = 60 mmHg) were subjected to surgery. In 22 patients we have studied systolic pulmonary pressures by echocardiography-Doppler examination at 3-6 and 12 months from surgery. We noticed a decrease in pulmonary pressure values in all patients within 3 months from surgery (mean values pre-op 75 +/- 12.14; mean values post-op 42 +/- 11.26); within 6 months from surgery physiological values were reached in all patients (mean values 35 +/- 5.3). We have further divided patients in two subgroups: group A with pulmonary pressures superior or equal to 80 mmHg (mean values 90.00 +/- 17.32) and group B With pressure values between 60 and 79 mmHg (mean values 69.12 +/- 3.64). The first Doppler evaluation demonstrated a greater decrease in group B (mean values 37.8 +/- 6.5 versus 57 +/- 12.4); at the second control there was no statistically significant difference between the two groups (group A 36 +/- 5.48, group B 33.33 +/- 5.37). The only two operative deaths were caused by the pre-op cardiogenic shock. There has been no late mortality among survivors, 22 in NYHA Class I and 2 in NYHA Class II. Pulmonary hypertension decreases after surgery independently of the adopted procedure (replacement, repair or lysis). PMID- 8233014 TI - [Aneurysm of the popliteal artery. Prognostic and therapeutic criteria]. AB - Experience of the management of 45 patients affected by popliteal artery aneurysms is reported. All patients, but one, underwent operative treatment. The importance of immediate surgical reconstruction whenever a popliteal artery aneurysm has been detected is stressed. In fact the most frequent complication represented by sudden thrombosis of the aneurysmatic sac and/or by peripheral embolization is generally followed by an irreversible closure of the vascular run off, determining the high rate of failures (28% in our experience) in spite of a prompt surgical reconstructive procedure. PMID- 8233015 TI - [The role of chronic renal insufficiency in the prognosis of surgical interventions in subrenal abdominal aortic aneurysm]. AB - Chronic renal insufficiency is one of the most important factors governing the immediate and long-term outcome after aneurysm repair. A total of 484 patients with abdominal aortic aneurysm (A.A.A.) have undergone elective surgical treatment in our Institution during the last 5 years. Of these, we selected 60 patients; 30 with a normal serum creatinine concentration and 30 with preoperative renal insufficiency (serum creatinine concentration greater than 2 mg/dl). In this second group, 23 patients (76.6%) were affected by slight or median renal insufficiency, 5 patients (16.6%) were affected by severe renal insufficiency (creatinine concentration greater than 4.5 mg/dl), and 2 patients (6.6%) had complete renal failure with dialytic treatment from 1.5 and 2 years respectively. We analyzed postoperative renal function in all 60 patients. In the first group, only 6 patients (20%) showed a transient renal insufficiency, without mortality and morbidity. In the second group, postoperative complications and mortality tended to occur more frequently in patients with a severe renal insufficiency than in patients with slight or median insufficiency or complete renal failure. The present data suggest that dialytic treatment might be necessary in patients with severe renal insufficiency before aneurysm repair. PMID- 8233016 TI - [Buerger disease. Report of an case, unusual for its site and clinical manifestations]. AB - The diagnosis of Buerger's disease, like that of all arteritis, especially in the onset phase, can be difficult. The general criteria proposed by several authors, with the aim of codifying the verification of the disease, are sometimes too schematic and this collides with the existence of very variegated and polymorphous clinical situations, as well as totally atypical and unusual clinical patterns. On this subject we consider useful to report a case of Buerger's disease that presented some peculiar characteristics, the most important of which was the existence of an aneurysm of the left superficial temporal artery. PMID- 8233017 TI - Morphine suppresses DNA synthesis in cultured murine astrocytes from cortex, hippocampus and striatum. AB - To determine whether there are regional differences in the ability of opiates to affect astrocyte proliferation, the effects of morphine were examined in astrocyte-enriched cultures from striatum, hippocampus and cerebral cortex derived from newborn mouse brains. Cultures from each region were continuously incubated in media alone (controls), or in media treated with 1 microM morphine, 1 microM morphine plus 3 microM naloxone, or 3 microM naloxone alone. Before harvesting at 6 days in vitro, cultures were exposed to [3H]thymidine (0.24 mu CI/ml for 16 h). Thymidine-labeling index was determined autoradiographically in flat, polyhedral (type 1) glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes. Morphine significantly inhibited [3H]thymidine incorporation in astrocytes from all three brain regions, although regional differences in labeling indices were noted. The results show that opiates can intrinsically affect the proliferative rate of astrocytes from diverse brain regions. PMID- 8233018 TI - Serum levels of beta-carotene and other carotenoids in Parkinson's disease. AB - To elucidate the possible role of carotenoids in the risk for developing Parkinson's disease (PD), we compared serum levels of beta-carotene, alpha carotene and lycopene, measured by high performance liquid chromatography, of 61 PD patients using their spouses as the control group. The serum levels of these 3 carotenoids did not differ significantly between PD patients and control groups. There was no influence of antiparkinsonian therapy on serum carotenoids levels, and these did not correlate with age, age at onset, scores of the Unified Parkinson Disease Rating Scale or the Hoehn and Yahr staging in the PD group. These results show that serum carotenoids concentrations are apparently unrelated to the risk for developing PD. PMID- 8233019 TI - Distribution of hippocalcin mRNA and immunoreactivity in rat brain. AB - Distribution of hippocalcin in rat brain was analysed by in situ hybridization and immunohistochemical methods. Hippocalcin mRNA and immunoreactivity were expressed more intensely in the pyramidal cells of the hippocampus, intensely in the Purkinje cells of the cerebellum, moderately in the dentate granule cells and pyramidal cells of cerebral cortex layers II-VI and weakly in the large neuronal cells of the caudate-putamen. Some discrepancies in the localization of hippocalcin mRNA and immunoreactivity were noted in the mamillary nuclei, anterior part of the thalamus and the septal nuclei. In most cell types, hippocalcin immunoreactivity was localized in the cytoplasm and plasma membrane of cell bodies and dendrites. PMID- 8233020 TI - NMDA receptor antagonist CPP inhibits ethanol-evoked ascorbate release in the brain of freely moving rats. AB - The effect of the N-methyl-D-aspartate (NMDA) receptor antagonist, 3-(2 carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP), on the ethanol-evoked release of brain ascorbate (AA) was investigated in freely moving rats using an in vivo voltammetry technique. Ethanol (1.0 g/kg, intraperitoneal injection, i.p.) increased the release of brain AA in the nucleus accumbens and striatum. Pretreatment with CPP (1 nmol, intracerebroventricular injection, i.c.v.) not only completely reversed the ethanol-evoked AA release but also reduced the concentration of extracellular AA and kept it at low level 80 min after the ethanol administration in both brain areas. However, 80 min after the treatment with CPP per se only the level of AA in the nucleus accumbens was decreased. The results suggest that ethanol-evoked release of AA probably is a self-protective mechanism against the neurotoxicity induced by ethanol in the brain, and the NMDA receptor may play an important role in this mechanism. PMID- 8233021 TI - The mechanosensitivity of spinal sensory neurons following intraspinal injections of quisqualic acid in the rat. AB - The mechanoreceptive properties of rat spinal sensory neurons were evaluated in segments adjacent to those injected with the excitatory amino acid agonist quisqualic acid. Following survival periods of 7-36 days cells recorded in quisqualate injected animals had an increased level of background activity, increased sensitivity to mechanical stimuli, and an increase in the duration of afterdischarge responses. It is suggested that a central mechanism that alters the functional state of neurons may be responsible for the sensory abnormalities, e.g. allodynia and hyperalgesia, that occur following excitotoxic induced cell death associated with ischemic and traumatic spinal cord injury. PMID- 8233022 TI - Identification of L-dopa immunoreactivity in some neurons in the human mesencephalic region: a novel dopa neuron group? AB - The presence of L-3,4-dihydroxyphenylalanine (L-DOPA) immunoreactivity is reported for the first time in some neurons in the human mesencephalic region, using an immunohistochemical method with a newly raised, highly specific anti-L DOPA antiserum. We have found many L-DOPA-positive/dopamine (DA)-positive and a few L-DOPA-positive/DA-negative cell bodies in dopaminergic regions in the midbrain. The present results suggest the existence of more than one neuronal group of L-DOPA in the human mesencephalon. L-DOPA in one group is an intermediate metabolite for decarboxylation to DA and in another group may exist as an end-product. L-DOPA in the latter neurons could be a neuromodulator and/or neurotransmitter. Thus, we suggest that L-DOPA plays an important role besides being an intermediate of DA in the human mesencephalon. PMID- 8233023 TI - Lateral preoptic lesions void slow-wave sleep enhanced by uridine but not by muramyl dipeptide in rats. AB - We investigated the site of action of two sleep-inducing substances, viz., muramyl dipeptide (MDP) and uridine. Localized electrolytic lesions were made bilaterally in the lateral preoptic hypothalamus (LPO) in rats and nocturnal 10-h i.c.v. infusions of MDP and uridine were performed before and after the LPO lesions. MDP increased only slow-wave sleep (SWS) in both intact and LPO-lesioned rats. Uridine promoted both SWS and paradoxical sleep (PS) before the LPO lesions whereas it increased only PS after the lesions. These results suggest that the LPO is crucial for SWS-promoting action of uridine but not MDP. PMID- 8233024 TI - Arterial baroreceptor information affects striatal dopamine release measured by voltammetry in rats. AB - The effects of altering the arterial blood pressure on the extracellular levels of dopamine in the corpus striatum, measured using nafion-coated C fiber electrodes combined with differential pulse amperometry, were assessed in urethane-anesthetized rats. The striatal dopamine release was increased by increasing the carotid blood pressure with phenylephrine injection but decreased by decreasing the carotid blood pressure with bilateral carotid occlusion. The data indicate that baroreceptors inputs affect the striatal dopamine release in rats. PMID- 8233025 TI - Visualization of G(o)alpha subtype guanine nucleotide-binding regulatory protein in the medulla oblongata and pons of the rat. AB - This study reports on the visualization of G(o) protein in the medulla oblongata and pons of Sprague-Dawley rats, using a commercially available selective polyclonal antiserum against a 39-kDa protein corresponding to the alpha subunit of G(o). Specific immunofluorescent-staining was widely distributed in neuropils and neuronal cell bodies but displayed regional heterogeneity. The immunoreactive product exhibited a punctate appearance and was most pronounced in the cell membrane and cytoplasm of the perikarya and proximal dendrites whereas the nucleus remained unstained. Positive immunoreactivity of G(o)alpha was present in nucleus reticularis gigantocellularis, nucleus reticularis paramedialis, nucleus tractus solitarii, nucleus nervi hypoglossi, nucleus raphe pallidus, locus coeruleus, nucleus tractus mesencephali nervi trigemini and nucleus cuneatus. These results provided anatomic evidence to support the participation of G(o) protein in both sensory and motor functions that originate from the medulla oblongata and pons of the rat. PMID- 8233026 TI - Induction of true precocious puberty by neonatal treatment with danazol in female rats. AB - Female rats at 5 days of age were given a single subcutaneous injection of varying doses of danazol or vehicle. Rats treated with 300 micrograms of danazol showed significant (P < 0.01, respectively) advancement in the day of vaginal opening (37.50 +/- 0.49 days vs. 25.20 +/- 0.19 days; mean +/- S.E.M.), first estrus (38.19 +/- 0.62 days vs. 29.10 +/- 0.26 days) and onset of estrous cycle (49.63 +/- 1.30 days vs. 37.07 +/- 1.50 days). All rats treated with danazol showed 4- or 5-day estrous cycle in the adulthood. Rats treated with 300 micrograms of danazol at 5 days of age discharged luteinizing hormone in the late stage of first proestrus. These results demonstrate that neonatally administered danazol produces the true precocious puberty in female rats, and this sexual precocity rat represents a unique model for analysis of the onset of puberty. PMID- 8233027 TI - Stimulatory effect of Saireito on proopiomelanocortin gene expression in the rat anterior pituitary gland. AB - The effect of administration of Saireito, a Saiko agent, via a stomach cannula on adrenocorticotropin (ACTH) release and gene expression of proopiomelanocortin (POMC), the precursor for ACTH, in the anterior pituitary, as well as on the corticotropin-releasing factor (CRF) in the hypothalamus, was examined in pentobarbital anesthetized rats. Saireito decreased the hypothalamic CRF level due to an early release of CRF and stimulated ACTH release and POMC gene expression but did not increase CRF gene expression. These results suggest that Saireito does not stimulate CRF gene expression, although it does stimulate CRF release, which in turn stimulates POMC gene expression in the anterior pituitary and ACTH release. PMID- 8233028 TI - Rostrocaudal changes in neuronal cell size in human lateral vestibular nucleus. AB - A cytoarchitectonic and morphometric study of the human lateral vestibular nucleus (LVN) is presented. In sagittal sections, the LVN appears as a triangular cell group rostrally located near the motor trigeminal nucleus and caudally near the vestibular root. The estimated volume is 13.49 mm3 with a neuronal population of 25,046 cells and 1855 neurons/mm3 in density. The average neuronal cross sectional area changes from a minimum caudally (380.02 +/- 7.23 microns 2) to a maximum rostrally (825.16 +/- 25.10 microns 2). Four types of neurons can be observed: small (< 200 microns 2), medium (200-500 microns 2), large (500-100 microns 2) and giant or Deiter's cells (> 1000 microns 2). The small and medium cells constitute 62%, large cells 26% and the giant cells only 12% of the neuronal population. PMID- 8233029 TI - Erythrocyte nuclei resemble dying neurons in embryonic dorsal root ganglia. AB - Cell death or apoptosis is regarded as an important feature of mammalian neural development, but the evidence for this generalization depends on the assumption that cell death can be clearly recognized. The usual profile of a dying neuron is a deeply stained pyknotic homogeneous sphere. In this paper we present evidence that such profiles in embryonic rat T6 and L4 dorsal root ganglia are not dying neurons but rather nuclei of immature red blood cells. This observation, combined with recent work showing that the methods previously used for counting normal or dying neurons are biased, indicates that the classic work establishing the importance of apoptosis needs to be repeated. PMID- 8233030 TI - Postnatal ontogeny of the rat brain neurotensin receptor mRNA. AB - Total RNA was purified from rat forebrain at different postnatal ages and analyzed by Northern blot using a specific neurotensin receptor RNA probe. The rat neurotensin receptor mRNA was present in high amount during the first 10 days of life. Thereafter, it rapidly decreased and was undetected after 20 days. [3H]neurotensin binding experiments performed on the same tissues indicated that the total amount of neurotensin receptors increased during the first week and was maximal between day 7 and day 10. This plateau was followed by an important loss (70%) of neurotensin receptors. These results indicate that an important reduction in the genetic expression of the neurotensin receptor after day 10 may probably account for the [3H]neurotensin binding profile observed in rat forebrain during the postnatal ontogeny. PMID- 8233031 TI - Expression of c-fos in brain subcortical structures in response to nauseant lithium chloride and osmotic pressure in rats. AB - Immunohistochemistry was used to map c-fos expression in rats to investigate the neural substrates that mediate the emetic action of lithium chloride and the effect of osmotic pressure. Solutions of 3% lithium chloride or 4.14% saline, isotonic to each other, as well as 0.65% lithium chloride or 0.9% saline, also isotonic to each other, were administered intraperitoneally (3 ml/kg) in rats. Both lithium chloride and osmotic pressure enhanced c-fos expression in the nuclei of the solitary tract, the paraventricular nuclei and supraoptic nuclei of the hypothalamus, and in the amygdala. This suggests that these brain structures might be the sites where the autonomic, neuroendocrine and behavioral responses elicited by lithium chloride and osmotic pressure are integrated. PMID- 8233032 TI - In situ hybridization of GAD mRNA in monkey and human brain: quantification at both regional and cellular levels. AB - GAD mRNA was detected in human and monkey brain postmortem by in situ hybridization with an [35S]-labelled copy RNA corresponding to a 2.7-kb fragment of the coding region of human GAD mRNA. A characteristic and reproducible pattern of hybridization was obtained with the anti-sense, but not the sense probe in both monkey and human brain. Microscopic examination of tissue sections showed that only neuronal perikarya, not glial cells, were labelled. The data confirm the heterogeneity of GAD mRNA distribution reported in rodent brain and non-human primate brain. PMID- 8233033 TI - Influence of barium on rectification in rat neocortical neurons. AB - The effect of low concentrations of barium on the membrane properties of rat neocortical neurons was studied in vitro. Potassium currents were examined using single-electrode current- and voltage-clamp techniques. Neurons responded to bath application of barium (10-100 microM) with a membrane depolarization associated with an increase in input resistance. Under voltage clamp conditions, an inward shift in holding current was observed. The effects of barium were rapidly reversible upon washing and persisted in the presence of TTX. The equilibrium potential for the barium-induced inward current was near -110 mV, suggesting that barium inhibited a tonically active potassium conductance. Measurements of current-voltage relationships indicated an inward rectification of this conductance between -50 and -130 mV. These results provide strong evidence that barium blocks a persistent potassium 'leak' current in neocortical neurons that contributes to the resting potential of these cells. PMID- 8233034 TI - [3H]2-(4-phenylpiperidino)cyclohexanol (AH5183) binding to synaptosomes and subcellular fractions obtained from rat brain. AB - We investigated the binding of [3H]AH5183 (2-(4-phenylpiperidino)cyclohexanol) to rat brain synaptosomes and subcellular fractions. A high content of specific binding was observed in crude synaptosomes obtained from the striatum, cerebral cortex and hippocampus. The highest density of subcellular binding sites was observed in the synaptic vesicle-rich fraction. The affinities of AH5183 binding to crude synaptosomes and the synaptic vesicle-rich fraction were almost equivalent, but the density of binding sites was higher in the synaptic vesicle fraction. The present findings indicate that [3H]AH5183 binding to both synaptosomes and the synaptic vesicle-rich fraction is useful as a cholinergic marker, and that for quantitative studies, binding to the latter fraction is more preferable. PMID- 8233035 TI - Neuropeptide FF (FLQPQRFamide) and IgG from neuropeptide FF antiserum affect spatial learning in mice. AB - There is evidence indicating that the mammalian octapeptide FLQPQRFamide (F8Fa or neuropeptide FF, NPFF) is an endogenous modulator ('anti-opioid') of opioid systems. There is also substantial evidence implicating opioid systems in the mediation of spatial learning and memory. In the present study determinations were made of the effects of NPFF and IgG from antiserum against NPFF on the spatial performance of male mice in a water maze task, whereby over one day in six blocks of four trials the animals had to acquire the location of a submerged hidden platform using distal visual cues. Pretraining intracerebroventricular (i.c.v.) injection of NPFF-IgG, impaired spatial acquisition and retention, while i.c.v. administration of 1.0 microgram of NPFF marginally improved, and 10 micrograms of NPFF significantly reduced spatial acquisition. These results suggest that NPFF may have a modulatory influence on spatial acquisition. PMID- 8233036 TI - Weak magnetic fields change extinction of a conditioned reaction and daytime melatonin levels in the rat. AB - Altogether 100 albino rats were conditioned to flee after an acoustic stimulus (1000 Hz). During the following retention period (extinction of the reaction), half of the animals (test group) were exposed to a vertically directed, weak and extremely low-frequency magnetic field (10 Hz, 10(-4) T). Under the field's influence, the animals showed an increased retention of conditioned reactions and had an elevated level of serum melatonin compared with the animals of the control group. These effects disappeared when the investigation was performed in the dark. PMID- 8233037 TI - Purine release and inhibition of synaptic transmission during hypoxia and hypoglycemia in rat hippocampal slices. AB - Evoked synaptic potentials and purine efflux were measured simultaneously from rat hippocampal slices. Slices were exposed to hypoxia, to glucose-free medium, and to in vitro ischemia consisting of glucose-free, hypoxic medium. During exposure to hypoxia or the glucose-free condition, radiolabelled purine efflux increased and the evoked population spike declined. Synaptic potentials and purine efflux returned to baseline values after reintroduction of normoxic and normoglycemic medium. During exposure to in vitro ischemia, purine and adenosine efflux were greatly increased with the appearance of the anoxic depolarization. PMID- 8233038 TI - Reduction of phosphoinositide hydrolysis by L-amino-3-phosphonopropionate may be caused by the inhibition of synthesis of phosphatidylinositols. AB - L-Amino-3-phosphonopropionate (L-AP3), a putative antagonist of metabotropic glutamate receptors, inhibited the formation of [3H]inositol phosphates induced by (1S,3R)1-aminocyclopentane-1,3-dicarboxylic acid in rat hippocampal slices. The inhibition was accompanied by a decrease in the levels of [3H]phosphatidylinositols ([3H]PIs). Preincubation of slices with L-AP3 inhibited the incorporation of [3H]myo-inositol into PI fractions. The effects of L-AP3 was in contrast with those of a typical receptor antagonist, atropine; atropine inhibited carbachol-induced phosphoinositide hydrolysis, but the levels of [3H]PIs were not affected. These findings suggest that the inhibition of phosphoinositide hydrolysis by L-AP3 is not due to the receptor antagonism but may be caused by the inhibition of synthesis of PIs. PMID- 8233039 TI - Amygdala kindling rate is altered in rats with a deficit in the responsiveness of the hypothalamo-pituitary-adrenal axis. AB - Fisher and Lewis rats were amygdala kindled in the morning and in the evening. Fisher rats displayed a diurnal variation in the rate of kindling through stage 2 which did not exist in the Lewis rats. Lewis rats also take much longer to kindle through stage 2 and a subcutaneous corticosterone pellet accelerates this rate. These observations are consistent with the hypothesis that the hormones of the hypothalamo-pituitary-adrenal (HPA) axis, particularly the glucocorticoids, play a role in epileptogenesis. PMID- 8233040 TI - Complement gene expression in neuroblastoma and astrocytoma cell lines of human origin. AB - Activation of the complement system is believed to be involved in degenerative processes of certain neurological diseases, including Alzheimer disease. Recent data have shown that the mRNAs for these proteins can be detected in brain derived mRNA. In this study, C4 mRNA was detected by polymerase chain reaction (PCR) amplification of mRNA from the human neuroblastoma cell lines IMR32, SK-SH and SK-MC, and the human astrocytoma cell line U373MG, while C3 expression was detected in SK-MC, SK-SH and U373MG cells. The SK-MC and U373MG cells expressed mRNA for C9. The mRNA for C1qB could not be detected in any of these cell lines. PMID- 8233041 TI - Aging prolongs the stress-induced release of noradrenaline in rat hypothalamus. AB - A stress-induced increase in noradrenaline (NA) release was measured by intracerebral microdialysis in the hypothalamic paraventricular nucleus of freely moving Wistar-Kyoto rats at three different ages (6, 18 and 24 months). NA levels in 20-min dialysate samples were measured by high-performance liquid chromatography with electrochemical detection. Microdialysis sampling was done at the baseline during a 20-min immobilization stress and for the next 100 min. Basal NA release was not significantly different in the three age groups. The immobilization stress increased NA levels (247, 197 and 234% of the baseline for the 6-, 18- and 24-month animals, respectively) which was not significantly different in the three groups. In the two younger groups NA returned to the baseline in the first sample after the end of the stress (t = 40 min) whereas in the 24-month group it remained significantly higher for longer (until t = 60 min). Stress-induced release of hypothalamic NA thus appears to be prolonged in old rats. PMID- 8233042 TI - [Ca(2+)+Mg2+]-dependent ATPase activity in rat pineal gland. AB - [Ca(2+)+Mg2+]-dependent ATPase activity in the pineal gland of the rat was examined. The enzyme possesses an apparent Km (Ca2+) of 0.23 microM, and moderately high affinity for Mg2+ and ATP (Km = 53.2 microM and Km = 86.4 microM, respectively). The ATPase activity is sensitive to low concentrations (I50 approximately 1 microM) of vanadate, which specifically inhibits Ca(2+)-ATPase in the plasma membranes of the erythrocyte, cardiomyocytes and synapses. The calmodulin antagonist trifluoperazine reduced significantly Ca(2+)-stimulated, Mg(2+)-dependent ATP hydrolysis. The [Ca(2+)+Mg2+]-dependent ATPase in rat pineal gland exhibits very high affinity for Ca2+, is highly vanadate sensitive and appears to require calmodulin. The enzyme is similar to the Ca(2+)-ATPase of the erythrocyte, cardiomyocytes and synaptic plasma membranes. These new findings may help to elucidate the mechanisms of intracellular calcium homeostasis and the effect of the enzyme on the synthesis of melatonin in the pineal gland. PMID- 8233043 TI - Distribution of VIP-immunoreactive nerve cells and fibers in the human ileocecal region. AB - VIP-containing nerve cells and fibers in the human ileocecal region (pre junctional ileum, ileocecal and cecocolonic junctions, post-junctional cecum and colon) have been evaluated by immunocytochemistry. A high density of VIP-positive neurons and nerve fibers was found in all layers of the ileum. At all colonic levels examined and at both junctions, the percentage of VIP-containing cells was higher in the submucous plexus than in the myenteric plexus. At both junctions, the muscle wall was devoid of, and the myenteric plexus extremely poor in VIP positive nerve fibers and cells. These data suggest that motility of these junctions is not--or only to a minor extent--regulated in man by VIP-containing nerves, at variance with other gut sphincteric areas. PMID- 8233044 TI - Cerebrovascular and metabolic uncoupling in the caudate-putamen following unilateral lesion of the mesencephalic dopaminergic neurons in the rat. AB - Changes in local cerebral blood flow (lCBF) and local cerebral glucose utilization (lCGU) were assessed in dopaminergic primary target areas in the rat 6 weeks after unilateral lesion of dopaminergic neurons within the substantia nigra pars compacta (SNc) and adjacent ventrotegmental area (VTA) using 6 hydroxydopamine (6-OHDA). lCBF and lCGU were determined using the autoradiographic [14C]iodoantipyrine and [14C]2-deoxyglucose method. Dopaminergic deafferentation provoked a marked unilateral lCBF decrease in the dorso-lateral portion of the rostral caudate-putamen. The decrease in lCBF was not associated with significant changes in glucose metabolism. Thus, lesions of dopaminergic afferents to the caudate-putamen appear to provoke a sustained decrease in basal blood flow with unchanged local metabolic activity. PMID- 8233045 TI - Antagonization of fentanyl-induced muscular rigidity by denervation of the coerulospinal noradrenergic pathway in the rat. AB - The present study examined the effect of denervating the coerulospinal noradrenergic pathway on the muscular rigidity elicited by fentanyl in Sprague Dawley rats anesthetized with ketamine. We demonstrated that the dopamine-beta hydroxylase-positive nerve terminals arborizing on spinal motoneurons that innervate the sacrococcygeus dorsi lateralis (SCDL) muscle were significantly eliminated by DSP4 treatment. Unilateral microinjection of fentanyl (2.5 micrograms/50 nl) into the locus coeruleus of these animals also failed to evoke discernible increase in the electromyographic activity recorded from the SCDL muscle. These results lend further support for our previous finding that the coerulospinal noradrenergic neurotransmission is critically involved in fentanyl induced muscular rigidity. PMID- 8233046 TI - The effect of intrathecal administration of RP67580, a potent neurokinin 1 antagonist on nociceptive transmission in the rat spinal cord. AB - The effect of intrathecal application of the selective neurokinin 1 (NK1) receptor antagonist RP67580 and its enantiomer RP68651 was studied on the responses of dorsal horn nociceptive neurones to formalin in the rat. The first and second phases of the formalin response were inhibited by RP67580 in a dose related manner (1-10 micrograms), whereas RP68651 (5 micrograms) facilitated the second phase of the response. The same doses of RP67580 had minimal effects on the acute C-fibre responses. The NK1 receptor appears to play a role in prolonged nociceptive transmission in the spinal cord. PMID- 8233047 TI - Enteric nerve fibers of holothurians are recognized by an antibody to acetylated alpha-tubulin. AB - The distribution of immunoreactivity to 6-11B-1, a monoclonal antibody that labels acetylated alpha-tubulin, was studied in the radial nerve and intestinal system of holothurians. As shown previously for other species, this antibody recognizes cilia and nerve fibers in Holothuria glaberrima and Holothuria mexicana. Thus, anti-acetylated alpha-tubulin can be used as a marker for nerve fibers in the enteric nervous system. PMID- 8233048 TI - Anti-B-series ganglioside-recognizing autoantibodies in an acute sensory neuropathy patient cause cell death of rat dorsal root ganglion neurons. AB - To examine the cytotoxicity of a patient's serum with an acute relapsing sensory neuropathy syndrome, dorsal root ganglion neurons from young adult rats were cultured in the presence of the patient's serum which had an extremely higher titer monoclonal IgM antibody recognizing B-series gangliosides, GD2, GD1b, GT1b and GQ1b. By the addition of the inactivated patient's serum, the relatively larger cells died after undergoing of metamorphosis during several hours of culture, whilst the smaller cells survived. The IgM fraction isolated from the patient's serum showed similar cytotoxicity towards the neurons as the inactivated whole serum. No cytotoxicity was observed with the IgM fraction containing medium after it had been absorbed with ganglioside GD1b. The results suggested that the anti-B-series ganglioside-directed antibody is the causal agent for the human neurologic disease. PMID- 8233049 TI - Human retinal ganglion cell development in early prenatal period using carbocyanine dye DiI. AB - Early development of retinal ganglion cell morphology has been studied in human fetuses of gestational age ranging from 7-8 weeks to 19-20 weeks by retrograde labelling with a lipophilic fluorescent dye, DiI. The retinal ganglion cells with simple and relatively uniform appearance having few dendritic processes between 7 to 11 weeks of gestation show progressive growth and elaboration of dendrites at 17-18 weeks revealing some morphological variation in shapes. It is by 19-20 weeks of gestation that the three major ganglion cell types resembling the alpha, beta and gamma classes of cat ganglion cells are identifiable in the developing human retina. The dendrites of some ganglion cells exhibit varicosities, filiform processes and spines even as early as 10-11 weeks indicating a possibility of involvement in synaptic connectivity. PMID- 8233050 TI - Rundown of NMDA-receptor mediated currents is resistant to lowering intracellular [Ca2+] and is prevented by ATP in rat spinal dorsal horn neurons. AB - Intracellular regulation of NMDA-receptor-mediated currents in cultured rat spinal dorsal horn neurons was investigated by means of simultaneously recording whole-cell currents and intracellular Ca2+ concentration ([Ca2+]i). During recordings in which EGTA (11 mM) was used to buffer intracellular Ca2+, NMDA currents showed 'rundown'; the amplitude of the currents gradually declined to a stable level approximately 50% of the initial level within 15 min of the beginning of recording. In these experiments, the level of [Ca2+]i decreased rapidly once whole-cell recording was attained and baseline [Ca2+]i remained below 100 nM. Each NMDA current was associated with a transient increase in [Ca2+]i which was prevented when BAPTA (30 mM) was substituted for EGTA. However, inclusion of BAPTA in the intracellular solution failed to affect the rundown of the currents. In contrast, including Mg-ATP (4 mM) prevented the rundown of NMDA currents and resulted in an increase in the current amplitude. Thus, our results indicate that rundown of the NMDA currents is not due to raised [Ca2+]i and are consistent with regulation of NMDA currents by phosphorylation. PMID- 8233051 TI - Actions of capsaicin on mouse dorsal root ganglion cells in vitro. AB - The effects of capsaicin were investigated on different populations of dorsal root ganglion cells in the in vitro mouse spinal cord-dorsal root ganglion preparation using intracellular electrodes. Dorsal root ganglion cells were characterised by the conduction velocity of their propagated action potential evoked by electrical stimulation of the dorsal root, and by the shape of their action potential. All cells with C-fiber characteristics (conduction velocity < 0.6 m/s; broad action potential with shoulder on the descending slope) were depolarised and generated action potentials when capsaicin (100-700 nM) was added to the bathing solution for 30 s. At these concentrations the membrane potential of DRG cells with myelinated fibers (conduction velocity > 2.0 m/s) was unaffected. Concentrations of capsaicin of 1.0-5.0 microM depolarised 50% of cells with conduction velocity > 10 m/s. During the depolarization of the membrane no action potentials were generated. In 50% of the capsaicin-sensitive neurons with conduction velocity faster than 10 m/s there was an initial hyperpolarization. Electrical stimulation of the dorsal root failed to evoke action potentials during the depolarization in 38% of the DRG cells with myelinated fibers and in all C-fibers tested within 10 min of the onset of the capsaicin effect. Passive depolarization of the membrane by intrasomal current injection mimicked the conduction block in neurons with large myelinated fibers. These observations confirm that capsaicin applied directly to the dorsal root ganglion affects, in a dose-dependent manner, both myelinated and unmyelinated primary afferents with a higher potency for C-neurons. Capsaicin evoked action potentials in C-neurons but not in neurons with myelinated fibers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233052 TI - Correlation between electroconvulsive seizure and HSC70 mRNA induction in mice brain. AB - The effects of electroconvulsive seizure and anti-convulsant drugs on induction of mRNA of heat shock protein were studied in mouse brain. Electrical shock induced mRNA of heat shock cognate protein (HSC70), but not heat shock protein (HSP70) mRNA. The induction was maximum 1 h after the ECS and continued for several hours, followed by long-lasting depression. Diazepam slightly prevented the ECS, but strongly attenuated the induction of HSC70 mRNA. Whereas phenytoin, which blocked the seizure, did not decrease but delayed the induction of HSC70 mRNA. The present results suggest that HSC70 mRNA level is increased with the ECS and that the induction level did not necessarily correlate the severity of the seizure. PMID- 8233053 TI - Circadian rhythmicity in the GABAergic system in the suprachiasmatic nuclei of the rat. AB - The participation of GABAergic mechanisms in the regulation of circadian rhythmicity by the suprachiasmatic nuclei (SCN) has been suggested from different lines of evidence. Little is known, however, whether GABA synthesis, release, uptake or content within the SCN may show a circadian pattern. The present results show that the activity of the GABAergic system within the SCN region of the rat exhibits circadian rhythmicity, which is manifested by correlative changes of the GABA content and the glutamic acid decarboxylase activity under the light/dark cycle, and by changes in the GABA content in animals kept under constant darkness. PMID- 8233054 TI - Localization of rat brain binding sites for [3H]tomoxetine, an enantiomerically pure ligand for norepinephrine reuptake sites. AB - The distribution of binding sites for the potent inhibitor of norepinephrine (NE) reuptake, [3H]tomoxetine, was examined in rat brain using quantitative autoradiography. Scatchard analysis of [3H]tomoxetine-binding to slide-mounted sections of rat forebrain indicated that the ligand bound to two sites, a high affinity site with a Kd of 0.29 nM and a lower-affinity site with a Kd of 16 nM. Pharmacological characterization of this high-affinity site was consistent with labelling a NE-uptake site in brain. Autoradiographic localization of the binding sites for [3H]tomoxetine was performed at a ligand concentration of 1 nM representing the distribution of high-affinity sites. The radioligand bound with a distribution of binding sites that was consistent with the known distribution of NE-containing neurons. The highest levels of binding were seen in regions, such as the locus coeruleus, bed nucleus of the stria terminalis, anterior ventral nucleus of the thalamus and the paraventricular nucleus of the hypothalamus. Low levels were seen in regions such as the caudate-putamen, ventral tegmental area and zona reticulata of the substantia nigra, where NE containing neurons have been reported to be low. Binding to all these sites was inhibited by 1 microM desipramine which produced autoradiograms with a uniform nonspecific binding. These results indicate that low concentrations of [3H]tomoxetine can be used to localize and characterize NE-binding sites. Further study will be necessary to determine the nature of the low-affinity binding site. PMID- 8233055 TI - Diverse modulation by systemic lidocaine of iontophoretic NMDA and quisqualic acid induced excitations on rat dorsal horn neurons. AB - The effects of systemic lidocaine (3-4 mg/kg) on the responses of 60 wide dynamic range neurons (WDR) to iontophoretically applied N-methyl-D-aspartic acid (NMDA) and quisqualic acid (QUIS) were studied in anesthetized, paralysed rats. The results show that lidocaine induced (i) potentiation of the NMDA excitation, reversible by 7-chloro-kynurenate (7-Cl-KYNA), a selective antagonist of the glycine binding site on the NMDA receptor; (ii) reduction of the QUIS excitation, reversible by strychnine (STRYCH), a glycine antagonist at its receptor. These findings, supporting a glycine-like action of lidocaine, are discussed together with data on the role of excitatory amino acids (EAAs) and the analgesic effect of lidocaine on neuropathic pain. PMID- 8233056 TI - Magnesium and ketamine attenuate cognitive dysfunction following experimental brain injury. AB - We evaluated the therapeutic effects of two noncompetitive antagonists of the N methyl-D-aspartate (NMDA) receptor, MgCl2 and ketamine, both individually and together, on cognitive dysfunction observed following parasagittal fluid percussion (FP) brain injury in the rat. Using a modified Morris water maze technique, we found significant attenuation of post-traumatic memory dysfunction in animals treated with either MgCl2 (125 mumol) or ketamine (4 mg/kg) (P < 0.005). Combined MgCl2 and ketamine treatment also preserved memory function (P < 0.005), with no apparent additive effect. PMID- 8233057 TI - Distribution of herpes simplex virus DNA in the brains of human long-term survivors of encephalitis. AB - We have examined the distribution of type 1 herpes simplex virus (HSV-1) DNA in the brains of 8 humans surviving for between 4 months and 17 years after acute encephalitis. Histological examination showed neuronal loss and gliosis largely confined to the temporal and frontal lobes, typical of HSV-1 encephalitis. There was a widespread persistent inflammatory infiltrate, present in both the cerebrum and brainstem. Viral DNA was detected in paraffin sections of formalin-fixed paraffin-embedded autopsy brain by use of the polymerase chain reaction to amplify a fragment of the HSV-1 thymidine kinase gene. HSV-1 DNA was amplified from the cerebrum in 6/8 cases and from the brainstem in 4/8 cases. These findings suggest the possibility that HSV-1 may persist within the human central nervous system after acute herpes simplex encephalitis. The distribution of the viral DNA correlates better with that of the persistent inflammatory infiltrate than with the destructive lesions of the acute encephalitis. PMID- 8233058 TI - GABA control of LHRH release is dependent on the steroid milieu. AB - The effect of GABA in controlling luteinizing hormone releasing hormone (LHRH) release from isolated hypothalamic fragments containing the mediobasal hypothalamus, suprachiasmatic nucleus, and the preoptic area (MBH-SCN-POA) was tested under different hormonal conditions with an in vitro superfusion method. GABA significantly decreased LHRH release under conditions of low, chronic levels of estradiol (E2) treatment but had no effect on tissue obtained from animals in proestrus. This suggests that the inhibitory effect of GABA is only manifested during certain stages of the estrous cycle characterized by negative feedback and low E2. PMID- 8233059 TI - Inhibition of outer hair cell electromotility by sulfhydryl specific reagents. AB - Mammalian outer hair cells can change length at acoustic frequencies when they are electrically stimulated. It was postulated that these length changes depend on electromechanical transduction based on voltage dependent conformational changes in a membrane motor protein. In this report, we describe the effect of various sulfhydryl (SH)-specific reagents on the OHC electromotility. p Chloromercuriphenylsulfonate (pCMPS), in addition to other mercurials that can react with well-protected SH-groups in proteins, inhibits this electromechanical transduction process. In contrast, N-ethylmaleimide and diamide, SH-reagents that only react with exposed SH-groups, showed no effect. These results suggest that one or more reactive SH-groups are present in a functionally important and protected region of the electromechanical transduction protein. Such reactivity can be utilized to identify and characterize this novel membrane motor. PMID- 8233060 TI - Use of mu-conotoxin GIIIA for the study of synaptic transmission at the frog neuromuscular junction. AB - We have looked at the effect of synthetic mu-conotoxin GIIIA, a selective blocker of muscle Na channels, on various parameters of synaptic transmission at the frog sartorius nerve-muscle preparation. We found that 5 microM mu-conotoxin consistently blocked muscle action potentials, but had no effect on nerve action potentials. The toxin also had no effect on the amplitude or frequency of miniature endplate potentials (MEPPs), on the amplitude or time course of endplate potentials (EPPs), or on stimulation-induced changes in EPP amplitude. The lack of an effect of synthetic mu-conotoxin GIIIA on transmitter release makes this toxin an invaluable tool in the study of neuromuscular transmission under conditions of normal levels of release. PMID- 8233061 TI - Species specificity of pharmacological characteristics of CCK-B receptors. AB - Novel CCK-B receptor antagonists, tetronothiodin and L-156,586, showed different affinities for CCK-B receptors in brain membranes from human, rat, guinea pig and mouse. [125I]CCK-8 bound to these membranes with a similar affinity. However, tetronothiodin was most potent in rat (IC50 = 3.6 nM), followed by guinea pig (96 nM), human (210 nM) and mouse (280 nM). L-156,586 bound with highest affinity to membranes from guinea pig (11 nM), and with lowest affinity to membranes from mouse (220 nM). These results suggest the existence of species specificity of CCK B receptors, and that these two compounds are useful tools for discrimination between these receptors. PMID- 8233062 TI - NADPH diaphorase activity is inhibited by EDTA in neurons but not in choroid plexus epithelium. AB - NADPH diaphorase histochemical staining was investigated in rat brain and choroid plexuses. All epithelial cells of the latter as well as some sparse neurons in striatum and cerebral cortex showed strong NADPH reaction product. While staining was homogeneous in neuronal cytoplasm, it was particulate in epithelial cells. Preincubation with EDTA (0.1 mM, 2h) prevented appearance of NADPH diaphorase reaction in neurons but not in choroid plexuses. These data show that in rat brain two forms of NADPH diaphorase are present; they are specifically localized in neurons and choroidal cells, respectively. PMID- 8233063 TI - Dantrolene protects against ischemic, delayed neuronal death in gerbil brain. AB - The effect was examined of dantrolene, a drug for malignant hyperthermia acting through preventing release of Ca from the ryanodine-type intracellular stores in muscle cells, on ischemic delayed neuronal death in field CA1 of gerbil hippocampus. Dantrolene (1.6 mM in concentration, 3 microliters each in volume), when administered bilaterally in the lateral ventricles 30 min after reperfusion after transient forebrain ischemia for 3 min at 37 degrees C, significantly protected against the neuronal death. It is proposed that the dantrolene sensitive (most likely, the ryanodine-type) intracellular Ca stores in CA1 pyramidal cells play a pivotal role in the development of ischemic neuronal damage. PMID- 8233064 TI - Presence of C-flanking peptide of neuropeptide Y(C-PON)-immunoreactive neurons in the olfactory cortex of the hedgehog (Erinaceus europaeus). AB - Although the presence of the neuropeptide C-terminal flanking peptide of neuropeptide Y, C-PON, has been described in the central nervous system (CNS) of mammals, to date there is no information related with its involvement in brain functions. An analysis of the location of C-PON in specific neuronal circuits of known anatomy and physiological action should provide light on its physiological role. The presence, distribution and morphology of C-PON-containing neurons in the olfactory cortex of the hedgehog was studied by immunocytochemistry. Immunoreactive neurons to C-PON were widely distributed in the three layers of the olfactory cortex of this primitive mammal. These neurons were medium sized and showed two or three immunostained, poorly branched, dendrites. In some positive neurons, a fine, beaded axon-like process was also immunostained. Although direct evidence of a physiological function of C-PON in the olfactory cortex of the hedgehog cannot be accurately stated from our findings, the morphology of C-PON neurons and their distribution in the deep cortical layers, where the majority of pyramidal neurons are located, suggest that this neuropeptide may play a role in the intrinsic neuronal circuitry of the relatively well-developed hedgehog paleocortex. A regulatory vascular role of some peptide-immunoreactive neurons can be inferred since occasional C-PON positive neurons have been located near blood vessels. PMID- 8233065 TI - Trypsin enhances sympathetic neuron-dependent plasma extravasation in the rat knee joint. AB - Perfusion of 6-hydroxydopamine through the rat knee joint causes an increase in plasma extravasation by activation of sympathetic neuron terminals. Similarly, the increase in plasma extravasation in the rat knee joint produced by the inflammatory mediator bradykinin is dependent on the sympathetic post-ganglion neuron. There is evidence that both 6-hydroxydopamine and bradykinin release a number of mediators, some of which appear to enhance plasma extravasation and some which inhibit it. We attempted to determine the nature of inhibitory factor(s) by co-infusing trypsin (which rapidly cleaves peptides) with 6 hydroxydopamine. We observed a marked enhancement of 6-hydroxydopamine-induced plasma extravasation by trypsin. This effect appeared to be specific to neurogenic plasma extravasation since trypsin alone had little effect on plasma extravasation and trypsin did not affect non-neurogenic plasma extravasation (that produced by platelet activating factor). Taken together, the data suggests that 6-hydroxydopamine not only releases mediators from the sympathetic neuron that produce plasma extravasation, but also an inhibitor(s) of plasma extravasation that is peptide in nature. PMID- 8233066 TI - The role of the striatum in organizing sequences of play fighting in neonatally dopamine-depleted rats. AB - Juvenile rats sustaining dopamine depletions by intraventricular injections of 6 hydroxydopamine (6-OHDA) as neonates were used to study the role of the striatum in controlling play fighting. As juveniles, the rats exhibited all the behavior elements typical of play fighting. However, they were more likely to use defensive tactics that shortened the playful contact between partners; and when contacting the partner, they were more likely to switch to other behaviors, such as allogrooming and sexual mounting, rather than continue with the play sequence. It is suggested here that the striatum is important for maintaining sequential organization of play fighting. PMID- 8233067 TI - Frequency-dependent plasticity of potentials evoked by repetitive stimulation of the lateral olfactory tract in rat olfactory cortex slices. AB - Habituation and sensitization were demonstrated in slices of olfactory cortex. One of the habituation properties, its dependence on the frequency of the lateral olfactory tract (LOT) stimulation, was tested. It was used in the frequency ranges (1, 10, 20, 30, 40, 50 and 100 impulses/s) which imitated the spontaneous activity of pyriform cortex (PC). Monosynaptic field potentials (FPs) were recorded in PC. Repeated stimulation with one of the frequencies of this range evoked different types of reactions in the PC populations of neurons. Three groups of neuronal reactions were distinguished. The first group of neurons manifested habituation (H-group), while the second one demonstrated sensitization (S-group). In the third group primary sensitization was observed, which then transformed into a short-term phase, when the amplitude of FPs did not change. After that the habituation was found to develop (S-H-group). In the H-group, it was more difficult to evoke habituation at the frequencies of 20-40/s and the number of stimuli in a trial of repeated stimuli (n) being equal to 10 and 150. In the S-group, it was more difficult to induce sensitization at the frequencies of 30-40/s and n equal to 10 and 150. This data proves that the development of habituation and sensitization depends on two factors at least: on the frequency of stimulation and on the type of excited synapse/cell population. Thus, in the PC neuronal populations, a property of habituation suggested by Thompson and Spencer [Psychol. Rev., 73 (1966) 16-43] (higher stimulation frequency causes more rapid and/or pronounced habituation) is not true for all ranges of frequencies. PMID- 8233068 TI - Immunocytochemical demonstration of neuropeptide Y and luteinizing hormone releasing hormone-immunoreactive structures in the organum vasculosum laminae terminalis of juvenile gilts. AB - Immunocytochemical investigations on the immature gilt organum vasculosum laminae terminalis showed extensive neuropeptide Y- and luteinizing hormone-releasing hormone-immunoreactive innervation of the organ. The luteinizing hormone releasing hormone-containing varicose fibers ran along the organum vasculosum laminae terminalis in close association with blood vessels. The nerve processes originating from well-stained luteinizing hormone-releasing hormone immunoreactive perikarya were distributed around the organum vasculosum laminae terminalis. The matrix of the organum vasculosum laminae terminalis was abundantly supplied by neuropeptide Y-immunoreactive varicose fibers. Numerous neuropeptide Y-immunoreactive terminals seemed to penetrate the ependymal lining of the organ. From these observations, it is concluded that there are favorable morphological conditions for secretion of neuropeptide Y into the cerebrospinal fluid of the third ventricle and release of luteinizing hormone-releasing hormone into fenestrated capillaries of the organ. PMID- 8233069 TI - O2 occlusion and cyanide induced immediate relaxation and contraction of murine skeletal muscle. AB - The acute changes of muscle tone and membrane current upon occlusion of oxygenation (O2 occlusion) were studied in vitro in mouse diaphragms. O2 occlusion immediately produced a contraction and a relaxation, respectively, in ryanodine- and high K(+)-contracted muscles while a biphasic change (an initial decrease then a late increase) of muscle tone was produced in muscles contracted with caffeine. The O2 occlusion effects were reversed after reoxygenation. CN- produced similar acute changes of muscle tone and abolished O2 occlusion effects. The O2 occlusion-induced relaxation in high K+ medium was converted into a contraction by 3,4-diaminopyridine and by low Cl- Tyrode's. O2 occlusion induced a small outward current and membrane hyperpolarization at a rate slower than the changes of muscle tone. Glybenclamide inhibited all of the changes induced by O2 occlusion. It is possible that the K+ and Cl- permeabilities of sarcoplasmic reticulum are highly sensitive to hypoxic challenge and related to the immediate changes of muscle tone after O2 occlusion. PMID- 8233070 TI - 1-Oleoyl-2-docosahexaenoyl phosphatidylcholine increased paradoxical sleep in F344 rats. AB - The effects of administration of 1-oleoyl-2-docosahexaenoyl-sn-glycero-3 phosphorylcholine (O-DH-PC), a kind of lecithin, and of glycerophosphorylcholine (GPC) on sleep were investigated in male F344 rats. Intracerebroventricular administration of O-DH-PC at a dose of 10 micrograms/rat induced significant increase in paradoxical sleep time and total sleep time in the following 24 h, while administration of GPC did not. Results suggest that O-DH-PC affects on neuronal mechanism relating to paradoxical sleep, and that the effect of O-DH-PC might be caused by fatty acid residues, rather than choline residue. PMID- 8233071 TI - Gene expression of high- (p140trk) and low-affinity nerve growth factor receptor (LNGFR) in the adult and aged human peripheral nervous system. AB - Steady-state mRNA levels and immunoreactive proteins for high- (p140trk) and low affinity nerve growth factor receptor (LNGFR) in the adult and aged human peripheral nervous system (PNS) were examined in autopsied material. trk mRNA expression was observed only in the sympathetic and dorsal root ganglia, while LNGFR mRNA was expressed widely through the PNS as well as non-neural tissues. Immunoreactive trk proto-oncogene product (p140trk) and LNGFR occurred in the perikarya of the subset of the sympathetic and dorsal root ganglion neurons, but only LNGFR immunoreactivity also occurred in the perineurium and the outer layer of the vessels. The spatial patterns of the trk and LNGFR gene expression in the adult human PNS were similar to those observed in the rat, mouse and chick, and their expression was well preserved in the aged. PMID- 8233072 TI - Response of neurons in the solitary tract nucleus, area postrema and lateral parabrachial nucleus to gastric load of hypertonic saline. AB - c-Fos was induced in the solitary tract nucleus (NTS), area postrema (AP) and lateral parabrachial nucleus (LPB) by the intragastric injection of hypertonic saline. The effect was bilateral and most dramatic in the NTS caudal to the facial nucleus and the AP, the areas receiving dense vagal afferent projection from the subdiaphragmatic structures. At least a part of these Fos-LI is considered to have been induced by the activation of the visceral osmoreceptors. PMID- 8233073 TI - Identification of androgen receptor in the rat spinal motoneurons. Immunohistochemical and immunoblotting analyses with monoclonal antibody. AB - Androgens play an important role in motoneuron growth, development and regeneration. We proved the existence of androgen receptor (AR) in the motoneurons of the rat spinal cord by the immunohistochemical stain and Western blotting. The possibility that AR protein in spinal cord is expressed in tissue specific form is proposed, being different from other androgen-dependent tissues. AR abnormality in X-linked spinal and bulbar muscular atrophy (SBMA) among a variety of motor neuron diseases were reported recently. Our study may give some clue to the AR abnormality leading to the degeneration of motoneurons. PMID- 8233074 TI - Excitatory and inhibitory effects of toluene on neural activity in guinea pig hippocampal slices. AB - To investigate the effect of toluene and its derivatives on neural activity, postsynaptic field potential (population spike, PS) of granule cells as well as antidromic potential (AP) and presynaptic fiber potential (FP) (perforant path) were recorded in the guinea pig hippocampal slices. Toluene at the concentration of 0.2 ng/ml to 20 micrograms/ml in the perfusion medium increased the amplitude of PS to 109-150%. Toluene also increased the amplitude of FP and AP, although the most remarkable enhancement was observed in the PS. However, toluene at the concentrations over 1000 micrograms/ml completely depressed the PS, whereas it increased the amplitude of AP to 130% of the original level. These results indicate that toluene has excitatory and inhibitory biphasic effects on neurotransmission in the hippocampal slices according to concentration applied. PMID- 8233075 TI - Are the fine-structural characteristics of mouse hippocampal mossy fiber synapses determined by the density of mossy fiber axons? AB - Heritable variation of mossy fiber synapses in hippocampal region CA3 was studied in the two inbred mouse strains C3H and CPB-K. Previous Timm studies had shown a larger mossy fiber projection in C3H mice. In contrast, synaptic boutons of CPB-K mice were larger in size and perimeter and were contacted by more dendritic spines than in C3H mice. These results point to an inverse relationship between the size of the mossy fiber projection and the number of spine synapses formed by an individual mossy fiber bouton. Thus, the fiber density of a projection may be crucial for the actual morphology of the synaptic contacts formed. PMID- 8233076 TI - Regional differences in the dopaminergic regulation of cyclic AMP formation within the rat nucleus accumbens: comparison with the striatal complex of the lizard Gekko gecko. AB - Effects of dopamine D2 receptor activation on the forskolin or D1 receptor stimulated formation of cyclic adenosine monophosphate (cyclic AMP) were investigated in tissue slices of two subregions of the rat nucleus accumbens and in the striatal complex of a lizard. Cyclic AMP production in the tissue was estimated by measuring the conversion of [3H]adenine in a superfusion system. Activation of a D2 receptor appeared to inhibit the D1 receptor agonists or forskolin stimulated formation of cAMP in the rostrolateral but not in the caudomedial part of the rat nucleus accumbens. In the striatal complex of the lizard the formation of cAMP was dramatically stimulated by forskolin, but only marginally by dopamine. Neither the forskolin, nor the dopamine stimulated cyclic AMP formation could be inhibited by activation of D2 receptors. These findings are compatible with previously obtained functional data indicating that especially the caudomedial part of the rat nucleus accumbens has much in common with the striatal complex of the lizard. PMID- 8233077 TI - Differential effect of chronic ethanol treatment on barbiturate and steroid modulation of muscimol-binding to rat brain cortex. AB - We report the differential alterations produced by chronic ethanol treatment on the modulation, by the barbiturate thiopental and the steroid 5 beta-pregnan-3 alpha-ol-20-one, of the binding of [3H]muscimol to membrane preparations from rat brain cortex. We found a clear barbiturate- and steroid-promoted enhancement of muscimol-binding to membranes in both control and ethanol-treated animals. However, the enhancements were higher in control animals, using the barbiturate, and in ethanol-treated rats, using the steroid, Bmax and Kd values were also differentially affected in control and ethanol-treated animals by the presence of the barbiturate or the steroid. PMID- 8233078 TI - Scyliorhinin-I and -II induce reciprocal hindlimb scratching in mice: differentiation of spinal and supraspinal neurokinin receptors in vivo. AB - Scyliorhinin-I amide (SCY-I) (selective for NK-1 and NK-2 receptors) or scyliorhinin-II amide (SCY-II) (selective for NK-3 receptors) were injected either spinally (i.t.; intrathecally) or supraspinally (i.c.v.; intracerebroventricularly) to mice. Following i.c.v. administration, SCI-I and SCY-II produced potent, dose-related reciprocal hindlimb scratching about equipotently (ED50 = 0.05 and 0.08 nmol, respectively). However, following i.t. administration, only SCY-I elicited greater than 50% response (ED50 = 0.07 nmol). Reciprocal hindlimb scratching is a behavioral response that has not been associated previously with neurokinins. These results might provide the first functional in vivo correlate for the differential localization of neurokinin receptor types within the mammalian central nervous system. PMID- 8233079 TI - Propentofylline (HWA285) inhibits the release of dopamine during transient ischemia and modulates its metabolism in rat striatum. AB - Propentofylline (HWA285) has been reported to protect neuronal cells through the inhibition of glutamate release during transient ischemia. We studied whether HWA285 inhibits dopamine (DA) release, and how HWA285 modulates DA metabolism in the rat model. HWA285 was perfused through a microdialysis probe placed in the rat striatum during 20 min transient ischemia. In rats perfused by HWA285, ischemic DA release was significantly inhibited, and DA metabolism showed better recovery in contrast with unperfused rats. PMID- 8233080 TI - Effects of tracheostomy on electrically induced vocalization in decerebrate cats. AB - Electrical stimulation delivered to the rostral pons induced well coordinated activity of the intralaryngeal muscles and the ventilatory muscles for vocalization in unanesthetized, decerebrate cats. The stimulation reset the normal respiratory rhythm and induced vocalization with a newly developed rhythm. Continuous opening of the tracheostomy decreased the breathing cycle during induced vocalization. Opening of the prepared tracheostomy during vocalization caused a sudden decrease in the subglottic pressure, but the expiration phase was not abruptly terminated. These findings suggested that vagal feedback from pulmonary stretch receptors plays a more significant role in the maintenance of the breathing cycle during electrically induced vocalization than the laryngeal feedback from the laryngeal mucosa. PMID- 8233081 TI - K(+)-stimulated 45Ca2+ flux into rat neocortical mini-slices is blocked by omega Aga-IVA and the dual Na+/Ca2+ channel blockers lidoflazine and flunarizine. AB - High-threshold neuronal voltage-sensitive Ca2+ channels (VSCCs) have been classified into at least three subtypes, including L, N, and P, based on biophysical and pharmacological criteria. We examined K(+)-induced 45Ca2+ flux into rat neocortical mini-slices to determine which of these subtype(s) might be involved in this phenomenon. Neither the L-type Ca2+ channel antagonist isradipine at 10 microM nor the N-type antagonist omega-conotoxin GVIA at 1 microM were effective antagonists of 45Ca2+ flux in this model. However, the P type Ca2+ channel antagonist, omega-Aga-IVA, blocked 70% of flux at 200 nM, with an IC50 of 17 nM, strongly implicating P-type Ca2+ channel involvement in K(+) stimulated Ca2+ entry into mammalian nerve terminals. About 30% of the flux response was resistant to the action of omega-Aga-IVA, suggesting that a still uncharacterized subtype of VSCC is involved in Ca2+ entry into mammalian nerve terminals. Both the omega-Aga-IVA sensitive and insensitive components of 45Ca2+ flux were blocked by the diphenylalkylpiperazines, lidoflazine and flunarizine (IC50 = 6.4 microM and 11 microM, respectively), which have dual Na+/Ca2+ channel blocking actions. PMID- 8233082 TI - Age-related loss of mRNA encoding adenosine A2 receptor in the rat striatum. AB - The distributions of adenosine A2 receptor mRNA and adenosine A2 binding sites were studied by in situ hybridization histochemistry and receptor autoradiography in young (3 months) and old (24 months) rat striatum and compared to that of enkephalin mRNA. In the old striatum, the levels of adenosine A2 receptor mRNA and adenosine A2 binding sites were reduced by 32% and 20%, respectively. This decrease is similar to the 24% decrease of enkephalin mRNA and to the previously reported 20% neuronal loss. This indicated that, in contrast to the dopamine D2 receptor, the reduction of adenosine A2 receptors in the rat striatum during aging is mainly due to the neuronal loss. PMID- 8233083 TI - Tonic GABA secretion of cultured rat hippocampal neurons rapidly transformed by Zn2+ into quantal release. AB - Different mechanisms of neurotransmitter secretion at synapses have been ascribed to quantal, transient signals [4-9] and to continuous, tonic activity [11, 21, 25]. Quantal transmission is imputed to be the release of transmitter packets, or quanta [6], assembled and stored in vesicles present in presynaptic terminals [2,8]. We report that 300 microM Zn2+ rapidly transformed tonic Cl- conductance mediated by the transmitter gamma-aminobutyric acid (GABA) into transient synaptic-like signals. After addition of Zn2+, the size of the elementary fluctuations in the Cl- current progressively increased while randomly activated Cl- channel kinetics remained unchanged and accounted for the bi-exponential time course of average transient decay. The results suggest that Zn2+ rapidly transforms tonic GABA secretion into quantal-like transient release. PMID- 8233084 TI - The noradrenergic innervation density of the monkey paraventricular nucleus is not altered by early social deprivation. AB - A series of neuroanatomic analyses have been undertaken to identify potential neuropathological changes seen in monkeys exposed to early social deprivation, which leads to psychopathology, inappropriate responses to stress and appetitive disorders. The animals used in this study were either socially reared or maternal and peer-deprived. Within this framework, the distribution and density of noradrenergic (and adrenergic) varicosities was assessed in the hypothalamic paraventricular nucleus of rhesus monkeys using dopamine-beta-hydroxylase immunohistochemistry combined with laser scanning microscopy. Quantitative analysis of dopamine-beta-hydroxylase-immunoreactive varicosity density within magnocellular and parvicellular regions revealed no significant differences between rearing conditions, suggesting that this chemically identified afferent input to the paraventricular nucleus was not affected by the early environmental insult of social deprivation. The apparent lack of vulnerability of the paraventricular nucleus to differential rearing conditions contrasts with the neuropathological changes observed in several discrete brain regions. PMID- 8233085 TI - Inositol 1,4,5-trisphosphate 3-kinase highest levels in the dendritic spines of cerebellar Purkinje cells and hippocampal CA1 pyramidal cells. A pre- and post embedding immunoelectron microscopic study. AB - Inositol 1,4,5-trisphosphate 3-kinase (InsP3 3-kinase) plays a crucial role in calcium homeostasis by regulating InsP3 levels. We have reported the highest concentrations of InsP3 3-kinase in the dendrites of cerebellar Purkinje cells and hippocampal pyramidal cells of the CA1 sector of the Ammon's horn. We here investigate its subcellular localization by pre- and post-embedding immunoelectron microscopic study. In both populations of neurons, the major structure expressing a high level of InsP3 3-kinase is the dendritic spines. PMID- 8233086 TI - Glycine effects on in vitro motor pattern generation in mouse spinal cord. AB - Bath applications of glycine typically inhibited electromyogram (EMG) activity in the gastrocnemius (G) and tibialis anterior (TA) muscles of neonatal mice, in vitro. Although rhythmic bursting occurred in response to glycine administration, cycle alternation between individual EMG bursts in G and TA muscles was not observed. That strychnine (a glycine 1 receptor antagonist) and cycloleucine (a glycine 2 receptor antagonist) could evoke motor rhythm, when given separately or in combination, confirms that glycine transmission is not required for motor pattern generation in mice. Strychnine application resulted in synchronized EMG bursting in G and TA muscles, suggesting that glycine 1 receptor activation does provide some reciprocal inhibition between the G and TA motor nuclei. PMID- 8233087 TI - Chronic neonatal N-methyl-D-aspartate receptor antagonism with MK-801 increases the number of corticospinal cells retained into adulthood in the rat. AB - The corticospinal tract elaborates and matures from an initial imprecise and wide spread projection to the more limited pattern seen in adult animals. This cortical projection to the spinal cord is refined through the elimination of 'inappropriate' axons. In the present experiments on the developmental shaping of corticospinal connections, an effect was observed after treatment with a non competitive antagonist of the N-methyl-D-aspartate (NMDA) receptor. Rat pups were exposed to the NMDA receptor antagonist MK-801 daily from the first to third postnatal week; littermates were injected with 0.9% saline and served as controls. At 4 weeks of age, animals were anesthetized, and Fast blue was inserted into the corticospinal tract at the 4th cervical segment of the spinal cord. The number of cortical neurons labeled with Fast blue was counted and compared between the two groups. MK-801 treated animals were smaller than their littermate controls. The results support the following conclusions: (1) the total number of cortical cells labeled from the cervical spinal cord placement was significantly increased (> 25%; P < 0.0005) in those animals who received MK-801 daily; and (2) the greatest increases of labeled cells were seen in the frontal and occipital cortices. These data emphasize the significance of the NMDA receptor in the shaping of central nervous system projections, and supports the use of the corticospinal projection as a model of development for glutamatergic connections. PMID- 8233088 TI - Ciguatoxin, extracted from poisonous morays eels, causes sodium-dependent calcium mobilization in NG108-15 neuroblastoma x glioma hybrid cells. AB - Measurement of intracellular Ca2+ concentration ([Ca2+]i) in cultured mouse NG108 15 neuroblastoma x glioma hybrid cells, using the fluorescent probe fura-2, revealed that 5-25 nM ciguatoxin (CTX) increased [Ca2+]i either in cells bathed in standard medium or after removal of external Ca2+ by a Ca(2+)-free medium supplemented with EGTA. Tetrodotoxin prevented the CTX increased [Ca2+]i suggesting that CTX-induced mobilization of intracellular Ca2+ depends on Na+ influx through voltage-gated Na channels. CTX-induced Ca2+ mobilization prevented subsequent action of bradykinin (1 microM) suggesting that CTX stimulates the inositol 1,4,5-trisphosphate-releasable Ca2+ store. PMID- 8233089 TI - Ganglioside GD3 enhances adherence of botulinum and tetanus neurotoxins to bovine brain synapsin I. AB - Tetanus toxin (TTx) and botulinum toxin serotype A (BTxA), preincubated with trisialoganglioside GT1b, adhere to proteins present on blots of bovine synaptosomal proteins. Differential solubilization and ammonium sulfate fractionation provided material enriched in two proteins that appeared to be adhered to most strongly by the labeled neurotoxins. After excision of the appropriate bands from blots of electrophoretically separated proteins, N terminal amino acid sequence analysis permitted identification of the proteins as synapsins Ia and Ib. Comparison of the effectiveness of different gangliosides at enhancing adherence of the neurotoxins to blots of synapsins Ia and Ib indicated that GD3 was most effective. PMID- 8233090 TI - Parvalbumin-immunoreactive dystrophic neurites and aberrant sprouts in the cerebral cortex of patients with Alzheimer's disease. AB - Parvalbumin-immunoreactive dystrophic neurites and aberrant terminal sprouts associated with senile plaques, together with preserved density of parvalbumin immunoreactive neurons, were found in the cerebral cortex of two patients with Alzheimer's disease, one of them familiar, in which a biopsy of the left frontal lobe was carried out for diagnostic and counselling purposes. These findings suggest that, although parvalbumin-immunoreactive cells are relatively resistant to degeneration in Alzheimer's disease, parvalbumin-immunoreactive neuronal processes can degenerate in some cases of Alzheimer's disease. PMID- 8233091 TI - NMDA-evoked adenosine release from rat cortex does not require the intermediate formation of nitric oxide. AB - Excitatory amino acids (EAAs) such as glutamate release the inhibitory neuromodulator adenosine from superfused rat cortical slices through the activation of both NMDA and non-NMDA EAA receptors. This study investigated the possibility that NMDA-evoked adenosine release may involve the intermediate formation of nitric oxide (NO). However, sodium nitroprusside did not evoke the release of adenosine, L-arginine did not augment and L-Nv-nitroarginine did not diminish NMDA-evoked adenosine release. It appears, therefore, that NMDA-evoked NO formation does not play a role in NMDA-evoked adenosine release in the cortex. PMID- 8233092 TI - Dependence of functional synaptic connections of hippocampal and neocortical neurons on types of memory. AB - Different types of memory are thought to be processed by different neuronal networks. The present study evaluated the functional synaptic connections among neurons by means of cross-correlation analysis when two types of memory, working and reference memory, were being processed in behaving rats. Some of the synaptic connections of neurons in the hippocampal CA1, CA3, dentate gyrus and auditory cortex functioned when the rat was performing only one of the memory tasks. Working memory needed more of these memory-type dependent synapses in the hippocampal regions than reference memory. PMID- 8233093 TI - GABAB receptor-mediated inhibition of forskolin-stimulated cyclic AMP accumulation in rat spinal cord. AB - GABA (30-1000 microM) and the GABAB agonist, (-)baclofen (10-100 microM), but not (+)baclofen, inhibited forskolin-induced cAMP formation in rat spinal cord slices. In contrast, GABA and (-)baclofen failed to enhance the stimulation of cAMP induced by noradrenaline in the same tissue, even though they both increased the response to noradrenaline in cerebral cortex slices. Neonatal capsaicin treatment, which reduces the density of GABAB binding sites in the cord, did not modify the effect of GABA or (-)baclofen on the forskolin-induced elevation of cAMP. (-)Baclofen and GABA inhibition of forskolin effects were insensitive to the GABAB antagonists CGP 35348 and CGP 36742 in the spinal cord. Since CGP 35348 antagonizes baclofen-induced antinociception, it seems unlikely that this effect stems from any change in cAMP within the spinal cord. PMID- 8233094 TI - Hypothalamic involvement in stress-induced hypocalcemia in rats. AB - Although hormonal regulation of blood calcium homeostasis has been intensively investigated in the peripheral organs, the involvement of the central nervous system in calcium regulation is still poorly understood. In the present study, we found that (1) bilateral lesions of the ventromedial nucleus of the hypothalamus (VMH), but not those of the paraventricular hypothalamic nucleus or the lateral hypothalamic area, eliminated immobilization (IMB)-induced hypocalcemia, and (2) electrical stimulation of the VMH decreased the blood calcium level. The results suggest that the VMH has a hypocalcemic function and plays a role in IMB-induced hypocalcemia. PMID- 8233095 TI - Behavioral activation by stimulation of a GABAergic mechanism in the preoptic area of rat. AB - The locomotor activity and grooming of conscious freely moving rats were recorded during a 60-min unilateral perfusion of the preoptic area with neuroactive compounds using the microdialysis technique. The GABA agonist, muscimol (10, 20 and 100 microM) induced a dose-dependent increase in locomotor activity and grooming which was attenuated by co-perfusion with the GABA antagonist, bicuculline (10 microM), and was blocked by systemic injection of haloperidol, a preferential dopamine D2 receptor antagonist (0.25 mg/kg). Muscimol-induced hyperactivity was associated with a simultaneous increase of striatal extracellular dopamine. These data suggest that the preoptic area is functionally linked with the extrapyramidal dopaminergic system possibly via GABAergic system. PMID- 8233096 TI - Muscarinic receptors mediate attenuation of extracellular acetylcholine levels in rat cerebral cortex after cholinesterase inhibition. AB - Muscarinic autoregulation of extracellular acetylcholine levels was investigated by microdialysis in the cerebral cortex of freely moving rats under basal conditions as well as following systemic administration of a reversible cholinesterase inhibitor. Atropine (2.2 mg/kg s.c. or 0.2 microM via the dialysis probe) did not affect basal extracellular acetylcholine levels in the cerebral cortex. However, it did potentiate the elevation of extracellular acetylcholine levels produced by a dose of systemic heptylphysostigmine which inhibited 25% of cortical and 40% of plasma cholinesterase activity. These observations suggest that the extracellular concentration of acetylcholine following moderate acetylcholinesterase inhibition is regulated through muscarinic receptors. PMID- 8233097 TI - Deficits in working but not reference memory in adult rats in which limbic seizures had been induced before weaning: implications for early brain injuries. AB - Male adult rats that displayed limbic seizures between postnatal days 18 and 21 after a single s.c. injection of Li followed 4 h later by a muscarinic agent were trained in a radial arm maze; they were compared with rats that had received the Li-pilocarpine (but had not displayed overt seizures) and to nonhandled controls. Only the rats that had displayed the (preweaned) seizures displayed significant impairment for working memory but not for reference memory. Light microscopy demonstrated histological evidence of earlier damage only within select thalamic structures that are directly associated with the amygdaloid-hippocampal complex. The results are compatible with the hypothesis that early seizures during the time of CA1 hippocampal maturation can produce long-term changes in the efficacy of short-term memory. PMID- 8233098 TI - The effects of cyclosporine on morphine-induced antinociception in diabetic mice. AB - The effects of cyclosporine on the antinociceptive effect of morphine were examined in diabetic mice. Sensitivities to the antinociceptive effect of morphine (5 mg/kg s.c.) in diabetic mice which had been pretreated with cyclosporine (30 mg/kg per day s.c.) for 14 days returned to those in vehicle treated nondiabetic mice. Naive mice which had been injected with the supernatant of spleen cell homogenate (SSCH) from vehicle-treated diabetic mice were less sensitive to morphine-induced antinociception. However, adoptive transfer of SSCH from cyclosporine-treated diabetic mice to naive mice had no effect on the recipients' antinociceptive sensitivities to morphine. These results suggest that the abnormal antinociceptive effect of morphine and the immune responses of diabetic mice may somehow be related. PMID- 8233099 TI - Evidence for D1 dopamine receptor-mediated modulation of the synaptic transmission from motor axon collaterals to Renshaw cells in the rat spinal cord. AB - The possible modulatory role of D1 dopamine receptors on the excitability of lumbar spinal Renshaw cells was studied in anesthetized rats spinalized at T4 level. Burst responses elicited by single electrical shocks to ipsilateral ventral roots L6 (frequency 0.5 Hz, stimulus width 0.1 ms) and spontaneous activity were recorded extracellularly using conventional 3 M KCl filled glass micropipettes. The specific D1 agonist SKF 38393 (0.5-1 mg/kg i.v.) enhanced Renshaw cell burst responses by 20-60% (n = 7) and increased their spontaneous discharge rate (n = 3). This effect was clearly antagonized by the specific D1 antagonist SCH 23390 (1 mg/kg i.v.) although SCH 23390 proved ineffective per se. We conclude that SKF 38393 induced facilitation was due to activation of the specific D1 receptors which could be the functional counterpart of the presynaptic D2 receptors described earlier by us in the same synapse. PMID- 8233100 TI - Fos induction in brainstem neurons by intravenous hypertonic saline in the conscious rat. AB - Experiments were done in conscious rats to investigate the effect of intravenous infusion of hypertonic saline on the induction of the protein Fos, in brainstem neurons. Neurons containing Fos-like immunoreactivity were observed in the caudal nucleus of the solitary tract (NTS), the caudal and rostral ventrolateral medulla, and parabrachial nucleus after an infusion of solutions containing 1.4 M NaCl. Little or no expression of Fos was detected in brainstem neurons after intravenous infusions of either physiological (143 mM) or hypotonic (106 mM) NaCl solutions. These data provide evidence for the involvement of brainstem structures in osmoregulatory functions and suggest that brainstem neuronal circuits that function in cardiovascular regulation may also be shared by those involved in body fluid homeostasis. PMID- 8233101 TI - The effects of thiamin and its phosphate esters on dopamine release in the rat striatum. AB - The effect of thiamin and its phosphate esters on dopamine (DA) release was examined in the rat striatum using an in vivo microdialysis. Intrastriatal administration of thiamin triphosphate (TTP) or thiamin diphosphate (TDP) induced DA release, but thiamin monophosphate (TMP) or thiamin did not show any change. In the absence of Ca2+ in the perfusate, TTP did not increase the DA release. omega-Conotoxin did not decrease the TTP-dependent DA release. These findings suggest that, in contrast to TMP and thiamin, TTP and TDP may play a specific role in DA release from nerve terminals. PMID- 8233103 TI - Probing blood supplies. PMID- 8233102 TI - GABAA-receptor immunoreactivity in the rat dorsal thalamus: an ultrastructural investigation. AB - The ultrastructural localization of GABAA-receptor (GABAA-R) immunoreactivity (ir) in representative nuclei of the rat dorsal thalamus was investigated using the monoclonal antibody 62-3G1 to the beta 2 and beta 3 subunits of the GABAA-R [8]. The pattern of distribution and the subcellular localization of ir were similar in all the thalamic nuclei examined, with the exception of the reticular nucleus that was unlabeled. The reaction product was present along somatic and dendritic plasma membranes of thalamic neurons and on their intracellular membranes. No labelling was observed in glial cells. The ir was present in areas of plasma membranes related and non related to terminals containing flat vesicles, and also on invaginated plasma membranes suggesting a recycling process of the receptor complex. The distribution and mismatches between GABA neurotransmitter and its receptor localization are discussed. PMID- 8233104 TI - Recent advances in laser treatment of benign cutaneous lesions. AB - Highly specific lasers provide an effective form of treatment for benign vascular and pigmented lesions. These lasers produce excellent cosmetic results while virtually eliminating adverse effects. With several laser treatments, clinicians can achieve dramatic results. PMID- 8233105 TI - Guide to pre- and postexposure rabies treatment. New Jersey State Department of Health. AB - Physicians are required by law to report all animal bites and human rabies postexposure treatments to a local health department. This information is extremely important to physicians and to New Jersey residents. The authors present recommendations for rabies pre- and postexposure treatment. PMID- 8233106 TI - Accurate bedside technique for differentiating wide-QRS complex tachycardias. AB - Patients undergoing open-heart surgery routinely receive temporary atrial pacing wires. The authors present a case report involving the use of these wires to differentiate the origin of a wide-QRS complex tachycardia. The technique, described in detail, should be utilized when appropriate. PMID- 8233108 TI - The new CLIA category: physician-performed microscopy. AB - The newest additions to the Clinical Laboratory Improvement Amendment (CLIA)-88 regulations are in the physician's interest. It is hoped this trend will continue as the Health Care Financing Administration (HCFA) reviews physician comments on the regulations. PMID- 8233107 TI - Teaching humanistic behavior: humanities study in the internal medicine residency. AB - Training in humanism is required in internal medicine residency programs, but teaching and evaluating humanism is difficult. Our program integrates the study of humanities into residency training for professional and personal development; betterment of communication skills; and understanding of ethics. PMID- 8233109 TI - Medical history: a New Jersey physician in the kingdom of Siam. AB - Charles Braddock, MD, lived and worked in Siam from 1902 to 1906. He was recognized as an expert in the management of epidemic diseases including smallpox, bubonic plague, cholera, and malaria. At his death in 1916, Dr. Braddock was noted to have a worldwide reputation. PMID- 8233110 TI - Pemphigus presenting as oral ulcers refractory to acyclovir therapy. AB - The authors present a patient with severe and persistent oral ulcerations despite acyclovir therapy. Because the patient initially had no skin lesions, the diagnosis of pemphigus was delayed. However, pemphigus commonly presents with oromucosal lesions before skin manifestations appear. PMID- 8233111 TI - Postsurgical radiation therapy for prophylaxis of heterotopic ossification. AB - Radiation therapy has been shown to be the most effective prophylactic measure against heterotopic ossification (HO) in surgically repaired hip fractures. Current research is seeking the minimum effective dose of radiation. The authors present four case reports and a review of the literature on HO. PMID- 8233112 TI - Physician hospital organizations: through lawyers' eyes. AB - A physician hospital organization may be a necessary step in the evolution of health care. However, many important decisions must be made in addressing the competing requirements of hospitals and physicians in the original design of the organization. PMID- 8233113 TI - Review article: transesophageal echocardiography. AB - Since 1990, we have performed transesophageal echocardiography (TEE) on 508 patients. This procedure has provided valuable information. The author reviews the indications and usage of TEE in various clinical settings and the need for further evaluation. PMID- 8233114 TI - An overview of post-stroke depression. AB - Post-stroke depression occurs in 22 to 60 percent of the 600,000 Americans affected each year by strokes. Without intervention, the prevalence and severity of depression for these patients is highest between six months and two years post stroke. PMID- 8233115 TI - Resident's note: entering a practice in the 1990s in New Jersey. PMID- 8233116 TI - Development of a motor nerve in the caudal fin of the medaka (Oryzias latipes). AB - The entire process of the development of the motor nerve to the middle interradial muscle (MIR muscle) in the tail region of the medaka (Oryzias latipes) was examined by light and electron microscopy. The motor nerve to the MIR muscle was observed to develop in a series of four steps. (1) Trunk-formation step: from stage 30 to stage 31, when the MIR muscle had not yet differentiated to the myotube stage, the motor axons starting from the caudal spinal cord, including spinal segment 27, formed a nerve trunk, the ventral caudalmost nerve (VCN). The VCN bypassed the place where the MIR muscle would differentiate and extended further caudad. (2) Wandering step: at stage 32, nerve processes of the motor axons (short branches) wandered from the course of the VCN. (3) Connection step: from stage 33/34 to stage 35, the mesenchyme cells differentiated into the MIR myotubes in the vicinity of the short branches, and the nerve and the muscle became connected. (4) Modification step: after stage 35, the morphological pattern of the motor nerve was modified by the growth and the shift of the short branches, and by the disappearance of the caudalmost part of the VCN. These results suggest that the complex pattern of the motor nerve in the adult fish is gradually constructed by multiple steps, each of which may have its own mechanism. PMID- 8233117 TI - Differential phasic modulation of short and long latency afferent sensory transmission to single neurons in the ventroposterolateral thalamus in behaving rats. AB - Single neurons were recorded in the forepaw area of the ventroposterolateral (VPL) thalamus of awake rats during rest and running behaviors. Locomotor step cycle dependent changes of the transmission of the short (4.0 +/- 0.1-10.1 +/- 0.3 ms, SEURs) and the long (10.2 +/- 0.2-26.0 +/- 2.1 ms, LEURs) latency somatic sensory responses were tested by generating post-stimulus time histograms of these neurons' responses to stimulation through electrodes chronically implanted under the skin of the forepaw. The magnitudes of firing during these responses were measured and normalized as percent increases over background firing. Times of footfall were determined through frame-by-frame analyses of video recordings and peri-footfall histograms were generated to differentiate a total of 40 VPL thalamic neurons into two types, footfall responsive (n = 21) and unresponsive (n = 19) neurons. Perifootfall gating patterns were determined for both types of cells. The SEURs and the LEURs showed significantly different phasic sensory modulation patterns across the locomotor step cycle. Major difference of the sensory modulations between footfall responsive and footfall unresponsive cells was noted during swing phase of the locomotor step cycle. In footfall responsive cells, the SEURs were suppressed most strongly just after footfall, while the LEURs were tonically suppressed during late stance and swing phases. The SEURs were disinhibited during the swing phase, while the LEURs were disinhibited during the middle stance phase. In footfall unresponsive cells, the LEURs were suppressed more strongly around footfall event than the SEURs were.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233118 TI - Pontine-induced generalized suppression of postural muscle tone in a reflexively standing acute decerebrate cat. AB - In a reflexively standing acute decerebrate cat, the pontine tegmentum was electrically stimulated and the effects on postural muscle tone and locomotor movements evoked by stimulation of the mesencephalic locomotor region (MLR) were studied. A stimulating microelectrode was placed systematically at 1-mm increments throughout the pons (H - 2 to H - 10) at levels ranging from P0.0 to P6.5 dorsoventrally and mediolaterally from LR0 to L4 or R4. Another stimulating microelectrode was placed in the physiologically identified MLR. Stimuli delivered to the dorsomedial regions of the pontine tegmentum (P3 to P4, LR1.5 to 2.5, H - 4 to H - 6) resulted in simultaneous and bilateral suppression of tonic activities in the neck, lumbar back, forelimb and hindlimb muscles. The pontine inhibitory sites corresponded to the medial area of the central tegmental field (FTC) and the central area of the gigantocellular tegmental field (FTG), bilaterally. Effects of pontine induced suppression on those muscles were stimulus frequency and stimulus intensity-dependent and the effects persisted even after termination of the stimulation. With concomitant pontine stimulation, MLR-evoked locomotor movements were suppressed along with prolongation of the forelimb and hindlimb step cycles. PMID- 8233119 TI - Extrastriate cortical neurons correlated with ocular convergence in the cat. AB - Responses of neurons in the lateral suprasylvian area to visual stimulation in association with ocular convergence were studied in eight alert cats trained to track a visual target moving in depth. Activities of 18 (3%) of 659 cells were related to ocular convergence. These 18 neurons were divided into two groups: activities of seven neurons (40%) correlated with peak velocities of convergence eye movement at both fast and slow target speeds (group I), while those of five neurons (30%) correlated with them only at faster target speed (group II). Activities of six other neurons correlated with peak velocity of ocular convergence at faster target speed but were not tested at slower speed. Activities of four group-I neurons (60%) did not or only weakly correlate with lens accommodation, while those of four group-II neurons (80%) correlated with peak velocity or amplitude of lens accommodation. It can therefore be concluded that the four group-I neurons are primarily related to ocular convergence. Other cells were either convergence-related, lens accommodation-related or both. It is suggested that these different types of neurons contribute in combination to optimal control of convergence eye movement. PMID- 8233120 TI - The membrane protein A5, a putative neuronal recognition molecule, promotes neurite outgrowth. AB - The A5 is an unique membrane protein that is expressed in the neurons of the visual system and general somatic sensory system of Xenopus laevis. We cultured retinal explants or trigeminal ganglion neurons on the monolayers of A5 expressing line cells obtained by transfection with the cDNA, and found that the A5-expressing transfectants promote neurite outgrowth for these A5-expressing neurons. The neurite outgrowth-promoting effect was inhibited by anti-A5 antibodies. While, the neurite outgrowth-promotion by the A5 transfectants was not observed for the vestibulocochlear ganglion neurons which lack the A5. These results indicate that the membrane protein A5 is a potent substrate for neurite extension of the A5-expressing neurons but not for the A5-negative ones, suggesting its involvement in specific neuronal interaction. PMID- 8233121 TI - A memorial address to the late Masao Udo. PMID- 8233122 TI - Structure and function of cerebral GABAA and GABAB receptors. AB - The receptor for GABA (gamma-aminobutyric acid), an inhibitory neurotransmitter in the brain, has been classified into GABAA and GABAB types. The GABAA receptor was purified by means of affinity column chromatography using benzodiazepine as an immobilized ligand. The results indicated that the GABAA receptor consists of several subunits and forms a GABA-gated Cl- channel, which is coupled with the benzodiazepine receptor. The molecular weight of the GABAA receptor complex was estimated to be approximately 300 kDa. Furthermore, cDNA cloning of GABAA receptor subunits was performed and the primary structure of these subunits was deduced. The results suggested that these subunits possess four transmembrane domains in their structure which are important for the formation of the Cl- channel. On the other hand, activation of GABAB receptors induced the inhibition of adenylyl cyclase activity and phosphatidylinositol turnover via inhibitory GTP binding proteins such as G(i) and/or G(o). The GABAB receptor was purified using baclofen affinity and immunoaffinity column chromatographies. It was confirmed that the purified GABAB receptor protein is about 80 kDa in its molecular weight. This protein is capable of inducing the inhibition of adenylyl cyclase when it is reconstituted with G(i)/G(o) protein in the phospholipid vesicle system. Currently available data indicate that GABAA and GABAB receptors in the central nervous system are distinct not only in terms of their molecules but also their signal transduction systems. However, the primary structure and synaptic localization of GABAB receptor molecules in the brain remain to be clarified. PMID- 8233123 TI - Functional roles of microglia in the brain. AB - It has been suggested that microglia, a type of glial cells in the central nervous system, play various important roles in normal and pathologic brains. In this article, we discussed the association or roles of microglia in injury and in brain diseases such as Alzheimer's disease, AIDS dementia complex, multiple sclerosis and ischemia. Furthermore, microglia-derived cytotoxic products and other secretory factors were summarized. In addition to the pathological aspects, secretory factors that showed neurotrophic effects were described with special reference to their physiological significance in the neuronal growth, neuronal function and regeneration processes. Accumulated evidence suggests that microglia are associated with not only brain pathology but also normal physiology in the brain. PMID- 8233124 TI - Hypercapnia and hypoxia which develop during retching participate in the transition from retching to expulsion in dogs. AB - The roles of arterial and central chemoreceptors in the transition from retching to expulsion during vomiting were studied. In spontaneously breathing decerebrate dogs, actual vomiting induced by activation of abdominal vagal afferents always consisted of retching and subsequent expulsion phases. Pulmonary ventilation almost stopped during the retching phase. Arterial blood CO2 tension gradually increased and reached a maximum near the time of the transition from the retching phase to the expulsion phase. Similarly, when end-tidal CO2 was maintained higher than 4.6 +/- 0.7% in paralyzed, artificially ventilated decerebrate dogs, stimulation of abdominal vagal afferents induced fictive retching and fictive expulsion, which were identified from the characteristic discharge patterns of the motor nerves to the costal and hiatal parts of the diaphragm, the abdominal muscles and the digastric muscle. However, only fictive retching occurred at an end-tidal CO2 of less than 3.7 +/- 0.7%. Although end-tidal CO2 was at a low level, fictive retching was followed by fictive expulsion when artificial ventilation was interrupted during the fictive retching phase and when sinus nerve afferents were stimulated. Even after sino-aortic denervation, fictive retching and subsequent fictive expulsion could be induced by stimulation of either vagal afferents or the solitary tract and nucleus, but the threshold level of end-tidal CO2 which enabled the induction of fictive expulsion increased after denervation. These results indicate that the activity of arterial and/or central chemoreceptor afferents must exceed some critical level to induce the transition from the retching phase to the expulsion phase. PMID- 8233125 TI - Laminar difference in tetanus-induced increase of intracellular Ca2+ in visual cortex of young rats. AB - Changes in intracellular Ca2+ evoked by electrical stimulation of the white matter were observed by means of microfluorometry with a Ca2+ indicator, rhod-2, in slice preparations of the visual cortex obtained from young rats. Tetanic stimulation at 5 Hz for 1 min induced a marked fluorescence increase, while single-shock stimulation did not induce a sizable increase in normal perfusate. The tetanus-induced increase took place in a column-like manner from layer VI near the stimulation site to layer II/III of the cortex, although it spread horizontally in layer II/III. The magnitude of fluorescence rise was largest in layer II/III of the cortex. Since N-methyl-D-aspartate (NMDA) receptors are known to exist only on neurons, the following results are taken to indicate that the fluorescent signal is derived mostly from postsynaptic neurons: Application of NMDA in the presence of tetrodotoxin induced a marked fluorescence increase with the same laminar bias as tetanic stimulation did, and the fluorescence increase by single-shock stimulation in Mg(2+)-free medium was almost completely blocked by an antagonist for NMDA receptors. These results support the hypothesis that input-associated entry of Ca2+ into postsynaptic neurons triggers processes for induction of long-term potentiation of synaptic efficacy. PMID- 8233126 TI - Reduction of GABA inhibition in Purkinje and cerebellar nuclei neurons in climbing fibre deafferented cerebella of rat. AB - GABA agonists were iontophoretically applied to Purkinje cells (PCs) of the cerebellar cortex and to neurons of the cerebellar nuclei (NCNs) in normal and in climbing fibre (CF) deafferented cerebella of rat. The experiments were performed one and three months after CF deafferentation obtained by total inferior olive destruction with 3-acetylpyridine. All control PCs were dose-dependently inhibited by GABA and muscimol and nearly all by baclofen. After CF deafferentation, the number of PCs sensitive to muscimol remained the same but the number sensitive to baclofen was greatly reduced one month later and almost absent after three months. The number of NCNs inhibited by GABA was slightly reduced one month after deafferentation compared to controls, but reduced to less than half three months after. Thus CF deafferentation of the PC leads to changes in postsynaptic sensitivity to GABA, the presumed inhibitory neurotransmitter, affecting GABAA receptors of the NCNs and GABAB receptors of the PCs. PMID- 8233127 TI - Fate and behavior of genetically labeled cerebellar cells after transplantation into mouse cerebellum. AB - A foreign gene coding the bacterial enzyme, chloramphenicol acetyltransferase (CAT), was introduced into a primary culture of the mouse cerebellar primordium by a retrovirus vector which harbors the neomycin-resistant gene. Following selection of the gene-transferred cells based on neomycin resistance, most of the selected cells expressed the CAT gene product as well as a marker for astrocytes, glial fibrillary acidic protein (GFAP), when examined immunocytochemically. These cells were transplanted into the adult mouse cerebellum, and the surviving cells were examined immunohistochemically by marking them with anti-CAT antibody. The distribution of CAT-immunopositive cells coincided with that of GFAP immunopositive cells observed in serial sections of grafted sites at 10 days after transplantation. Some of the transplanted CAT-immunopositive cells extended processes and exhibited the morphological appearance of fibrous astrocytes. Migration of the genetically labeled cells into the host molecular layer was also observed and the morphological plasticity of the differentiated primary cells was shown according to the grafted sites. These results indicate that stable marking of cells for grafting can be accomplished by retrovirus-mediated introduction of a foreign gene into the primary culture, and that the fate and behavior of the labeled donor cells can be analyzed immunohistochemically following transplantation into neural tissue. PMID- 8233128 TI - Using questioning as a teaching strategy. AB - Effective use of questioning as a teaching strategy can help nursing students draw on and apply acquired knowledge in new and unique situations. By considering the question's classification, environment, source, delivery, and response, faculty can improve their use of this important teaching strategy. PMID- 8233129 TI - Use of clinical journals to enhance critical thinking. AB - The clinical journal offers a valuable medium through which faculty can teach critical thinking. Both analysis and evaluation of the interactions and relationships between clinical practice, theory, and research can be greatly enhanced through careful structuring of clinical journal assignments. The authors discuss ideas about structuring clinical and journal assignments and strategies for giving feedback. PMID- 8233130 TI - Using algorithms to teach dosage calculation. PMID- 8233131 TI - Educational implications of community health preceptors' practice values. AB - Educators in baccalaureate nursing programs realize that clinical experiences in community health differ from those in acute care facilities. Competencies required in community settings must be identified to ensure that students have had opportunities to develop necessary skills before obtaining clinical experience. Such identification is also necessary for evaluating student competence in the clinical setting. The authors identify valued community health practice competencies that could be used by educators to evaluate students and help them prepare for clinical practice. PMID- 8233132 TI - Martha Rogers' model in associate degree education. AB - This article describes the first associate degree nursing program in the country to adopt Martha Rogers' science of unitary humans as the conceptual model for the program. The authors discuss the rationale for choosing the model, the curriculum design, the special challenges to implementation, and student and faculty reactions to the model. PMID- 8233133 TI - Caring relationships between nursing students and their patients. AB - The meaning of a caring experience between a nursing student and a patient was explored in this phenomenologic study. Twenty-two undergraduate students wrote an in-depth account of a caring experience they had with one of their patients. Using Van Manen's phenomenologic approach, the following five essential themes of a caring nursing student-patient experience emerged: authentic presence, competence, emotional support, physical comforting, and positive consequences. Implications for nurse educators are addressed. PMID- 8233134 TI - Bias against the nurse. PMID- 8233135 TI - Avoiding a type VI error in program evaluation. PMID- 8233136 TI - Healthcare for the homeless: RN to BSN clinical education. AB - Registered nurse students benefit from community health clinical placements in which they are able to effect change for the clients and the community in which they work. A nursing clinic in a soup kitchen for homeless people has demonstrated the effectiveness of student involvement in program development. Students and faculty work on the front line of life in direct care and advocacy for an underserved population. PMID- 8233137 TI - Can competency-based education survive the nineties? PMID- 8233138 TI - Nursing articulation in the state of Idaho. PMID- 8233139 TI - Nursing theories: perception of a student. PMID- 8233140 TI - Discovery of meaning through imagined experience, writing, and evaluation. AB - Writing is frequently referred to as a skill. Its value, however, is most often judged by what it produces, when, ironically, studying the process of writing may be the best way to improve the product. This teaching plan asks students to develop a meaningful written product while focusing on context, content, and process evaluation. It also provides a learning experience that has the potential to enhance empathy and caring among nursing students. PMID- 8233141 TI - Nurse-midwifery within a faculty group practice. PMID- 8233142 TI - A social survey: public perceptions of crisis in health care and responses to advanced nursing role in a Midwestern community. PMID- 8233143 TI - Quality, access, and clinical issues in a nurse practitioner colposcopy outreach program. AB - From 8/89 to 7/92 a NP-colposcopist performed 593 colposcopy examinations including cervical biopsies, endocervical curettages, and Pap smears. Procedures were performed in selected health departments in north-central Florida. This portable outreach program expanded services to a primarily rural, medically indigent population. Specimen results were 21.1% negative, 5.6% human papilloma virus (HPV) only, 37.3% CIN I, 17.4% CIN II, 11.9% CIN III, 2.4% CIS, 0.3% carcinoma, and 4% other. Many CIN lesions also included HPV, yielding an overall incidence of HPV at 62.7%. Using the Bethesda system, 42.8% were low grade lesions, 32.1% high grade. Fourteen cases of CIS and two of carcinoma (one invasive, one microinvasive) were found. An 87.7% colposcopic diagnostic acumen level is favorable compared with published physician studies. One third of undercalls were due to occult canal lesions. Significant risk factors were smoking (48.6%), first coitus at < 18 years old (74%), history of HPV (49.6%), and life-time sexual partners > 2 (61.3%). Holistic prevention enabled 209 non colposcopy problems. PMID- 8233144 TI - Preventing diabetic foot disease. AB - Diabetic foot disease results from two common pathologies: peripheral vascular disease and diabetic neuropathy. If these pathologies are not identified, ulceration may occur in the foot. Ulcers can lead to infection and finally amputation. This article discusses the components of the physical exam, current treatment for both pathologies, and client education. Because the two pathologies are similar in presentation, it is important for clinicians to distinguish between the two pathologies. Careful attention to the feet during the physical exam and assessment for symptoms can help distinguish the two pathologies. There are new treatments available for peripheral vascular disease to improve the client's circulation. Treatment of peripheral neuropathy is usually palliative, but can improve the client's quality of life. Through early identification and treatment of peripheral vascular disease and diabetic neuropathy, clients with diabetes can avoid ulceration. Client education is central to the treatment plan. PMID- 8233145 TI - Guidelines for primary care follow-up of premature infants. AB - This article provides general guidelines for assessing premature infants during their first two years of life. Statistics are presented to validate this population's increased risk of medical, neurologic and developmental problems. Developmental assessment and the need to use corrected age with standardized tools is explained. Immunizing preemies according to their chronological age is discussed. The following content is reviewed: growth parameters with head circumference and intrauterine growth retardation, nutrition and caloric requirements, necessary screenings with referral information, medical problems with apnea monitor management, and anticipatory guidance including the Vulnerable Child Syndrome. PMID- 8233146 TI - Encopresis: a study of treatment alternatives and historical and behavioral characteristics. AB - The historical, demographic and pre/post treatment behavioral profiles of a Health Maintenance Organization's (HMO) encopresis patients were studied. The efficacy of two maintenance treatments were also compared. Management included an initial bowel evacuation, high fiber diet, behavior modification program, and random assignment to maintenance on oral laxatives or conditional rectal cathartics. Of the 136 patients evaluated, 61 completed treatment (oral 24; rectal 37). Significant behavioral problems were identified. However, the majority (65%) of patients were managed without psychosocial intervention and successful completion of treatment significantly improved most behaviors. There were no significant differences in efficacy between the oral laxative or conditional rectal cathartic maintenance groups completing treatment. In 87% success continued 6 to 12 months post-treatment. PMID- 8233147 TI - Health care crises: nursing solutions. PMID- 8233148 TI - Affecting children's attitudes toward diabetes through camp. PMID- 8233149 TI - Survey of NP salaries in Maryland. PMID- 8233150 TI - More tips for a successful independent practice. PMID- 8233151 TI - How to analyze nurse practitioner licensure laws. AB - Nurse practitioners (NPs) are capable of analyzing the regulation of their practice by state authorities. The annotated nursing statute, the medicine and pharmacy statutes, regulations from those three boards and cases noted in the annotated statutes should be read. Nurse practitioner definitions of practice can be broadly written (making relicensure in a state with similar definition easier) or specific (requiring all states to adopt similar qualifications for endorsement, as is done now with basic RN licensure). Requirements for NP credentialing by nonstate entities have advantages and problems too. State granted monopoly of licensure is good for NP economics and bad for patient economics. Medical practice law is important to the NP. Pharmacy statutes are important even if the NP statute provides for prescribing. Revision of NP pharmacy law is becoming less necessary as more control of prescribing by guidelines is exercised on nurses and physicians alike. Rules for analysis of statutes are given. PMID- 8233152 TI - Pediatric sexuality: promoting normal sexual development in children. AB - Since the sexual revolution of the 1960s there has been an openness regarding sexual exploration that has resulted in an increase of sexually transmitted diseases and teenage pregnancies. Clinicians can mitigate the unhealthy results of such exploration through a therapeutic relationship with their patients. This article provides practical ways to approach and educate the pediatric patient and parent regarding normal sexual growth and development and the promotion of healthy, responsible sexual behavior. Using developmental theories as a foundation, sexuality is examined from birth through the adolescent years. PMID- 8233153 TI - Social constructivist approaches for brief, episodic, problem-focused family encounters. AB - Clinicians of many health care disciplines provide health care services to the family unit. Because of the multiple perspectives that individual family members bring to the health care encounter, family-focused primary health care presents special challenges for providers. The need to reconcile multiple individual family member perspectives requires that clinicians develop increasingly more sophisticated, effective, family intervention skills that can be used in busy primary care settings. Social constructivism can provide a means through which clinicians can quickly engage families in creating a consensus on health issues, thus leading to effective, efficient problem solving. This paper reviews the origins and principles of social constructivist thought and describes assessment questions and therapeutic stances that can be used to empower families to become active participants in the process of initiating their own health-related life changes. These approaches are useful for episodic, brief, problem-focused encounters with families in many primary care practice settings. PMID- 8233154 TI - Primary care screening and evaluation of hearing loss. AB - Approximately 20 million Americans suffer from some degree of hearing loss. Screening, evaluation, treatment and referral in the primary care setting can reduce the degree and adverse effects of the loss. By asking specific health history questions, the clinician can identify risk factors, symptoms and possible causes of hearing loss. Simple behavioral tests, tuning fork tests, audioscopy, audiometry and tympanometry help the clinician determine the presence, type and degree of hearing loss. Some causes of hearing loss can be treated in the primary care setting, but many require special testing, surgical intervention and rehabilitation. Resources for referral include otolaryngologists, audiologists and organizations for the hearing impaired; they can provide further testing, education and support to clients with hearing loss. PMID- 8233155 TI - Assessing pulse deficit. AB - Pulse deficit--the difference between the apical and peripheral pulse rates--can signal an arrhythmia. You'll need to monitor this deficit if your patient's pulse rhythm is irregular. PMID- 8233156 TI - Notifying the doctor by phone. PMID- 8233157 TI - A.I.D.S. patient. Finding contentment. PMID- 8233158 TI - Determining your elderly patient's nutritional health. PMID- 8233159 TI - Instilling eardrops. PMID- 8233160 TI - Managing the adult organ donor. PMID- 8233161 TI - Responding to pulmonary hemorrhage. PMID- 8233163 TI - Cost-effective teamwork. PMID- 8233162 TI - What you need to know about seizures. PMID- 8233164 TI - Measuring paste another way. PMID- 8233165 TI - What you can do to reduce increased I.C.P. PMID- 8233166 TI - We felt safe in Iowa ... until the explosion. PMID- 8233167 TI - Administering insulin through an indwelling catheter. PMID- 8233168 TI - Dropping the barriers between Jim and us. PMID- 8233169 TI - Myths & facts ... about insulin. PMID- 8233170 TI - Improving your listening skills. PMID- 8233171 TI - Six tips to boost morale. PMID- 8233172 TI - Something to remember. PMID- 8233173 TI - Bone mineral density in patients receiving suppressive doses of thyroxine for thyroid carcinoma. AB - AIMS: Studies of the effect of thyroxine therapy on skeletal integrity have given conflicting results; the reductions in bone mass reported by some have prompted recommendations that the prescribed replacement doses of thyroxine should be reduced. We have examined bone mineral density in a group of patients with differentiated thyroid carcinoma receiving high doses of thyroxine to suppress thyroid stimulating hormone (TSH). METHODS: The 44 patients (6 male, 38 female) had a median age of 49 years (range 27-75) with median duration of thyroxine therapy of 9.0 years (range 3 to 42) and mean dose of thyroxine 0.167 mg/day (range 0.125-0.3). TSH levels were chronically suppressed in 39 subjects. Bone mineral density (BMD) was measured by dual energy x-ray absorptiometry (DEXA) in all subjects at the femoral neck and lumbar spine and compared with previously established local reference ranges. RESULTS: There was no reduction in bone mineral density in the thyroxine treated group compared with the local reference population at both lumbar spine and femoral neck, and no correlation with duration of therapy. CONCLUSIONS: These negative findings, that thyroxine in suppressive doses does not significantly reduce bone mineral density in New Zealand patients suggest that thyroxine therapy alone is not a major risk factor for the development of osteoporosis. PMID- 8233174 TI - Ultraviolet protection without ultraviolet exposure? PMID- 8233175 TI - Education about death and dying at Otago University Medical School. AB - AIMS: To ascertain if medical training provides adequate preparation for students regarding; (a) coming to terms with death and dying, (b) communication pertaining to terminal illness and (c) symptom control of the dying. METHODS: A questionnaire was distributed to all undergraduates in years 3 to 6, of the Otago Medical School, including all three clinical schools. The students were asked to rate their perceived abilities in given clinical situations. RESULTS: There were 392 replies giving ratings on a scale from 0, for least confidence in ability, to 5, for most confidence in ability. A mean rating of 2.45 was given for preparation prior to cadaver dissection. A mean rating of 2.31 was given overall for communication skills pertaining to terminal illness with an average increase of only 0.49 between years 3 and 6, but 1.89 in the area of symptom control, with an average increase of 2.32 between years 3 and 6. Many students stated that extra teaching was required, primarily in areas relating to communication. CONCLUSION: A challenge exists for medical teachers to increase curriculum content in this area and suggestions are given. PMID- 8233176 TI - Erythema multiforme associated with an outbreak of Mycoplasma pneumoniae function. AB - AIMS: To document the association between erythema multiforme and Mycoplasma pneumoniae. METHOD: All cases of erythema multiforme presenting to the dermatology department of Waikato Hospital over the summer of 1992/3 were tested for mycoplasma serology. Trends in recorded cases of M pneumoniae infection were examined from data supplied by the virology department of Waikato Hospital. RESULTS: Four of seven cases of erythema multiforme were due to M pneumoniae infection. This was associated with a marked local increase in the number of M pneumoniae infections recorded in our hospital. CONCLUSION: Mycoplasma serology is an important investigation in the management of erythema multiforme, especially when associated with an outbreak of M pneumoniae infections. PMID- 8233177 TI - Case mix and outcome for patients with fracture of the proximal femur. AB - AIMS: To analyse the way in which case mix and outcome are related for fracture of the proximal femur in a base hospital. METHODS: All patients over age 60 admitted to Waikato Hospital during one year with fracture of the proximal femur were assessed for premorbid function and comorbidity and were followed to discharge. RESULTS: There was a low frequency of post operative morbid events. Post operative mortality was 3%. Average length of stay was 19.9 days. Increased age and disability, trochanteric fracture, residence in a rest home, previous hypertension or stroke, technical complication, simultaneous fracture of another limb and perioperative myocardial infarction were all associated with increased length of stay. CONCLUSION: Factors are described which will alert clinicians to people likely to have a long length of stay and may need formal rehabilitation. Continuous quality improvement techniques may be a way of improving services such as these further. PMID- 8233178 TI - Patient's medicines brought to hospital: an overlooked resource? AB - AIM: To quantify potential cost advantages and identify practical safeguards required for utilising patients own medicines while in hospital, and returning them on discharge. METHODS: All medicines brought in by patients in two wards of a geriatric assessment and rehabilitation unit at Auckland Hospital were examined by the pharmacist, and their suitability for re-issue assessed. Medicines were regarded as suitable for use where they could be positively identified, had been dispensed within 3 months of admission, or if packed in foil, provided the expiry date and manufacturer identification were on the foil. RESULTS: Medicines (260 items) totalling $2,976, assessed over a 6 month period, were regarded as suitable for use by the patient, with a mean value of $11.36 per patient. CONCLUSIONS: Patients own medicines used within the hospital with a unit-of-issue distribution system, and taken home by them on discharge, would provide appreciable savings for the hospital medicine budget and reduction in waste of the overall health dollar. PMID- 8233179 TI - The management of epilepsy in adults. PMID- 8233180 TI - Tropical cyclone Prema. PMID- 8233181 TI - Attitudes to alcohol: regulating the market. PMID- 8233182 TI - A chair of rehabilitation medicine at Otago? PMID- 8233183 TI - Dr Jean Pierre Allain and factor VIII. PMID- 8233184 TI - Post polio syndrome. PMID- 8233185 TI - Occupational overuse syndrome. PMID- 8233186 TI - Sexual abuse and mandatory reporting. PMID- 8233187 TI - Fluoridation and bones. PMID- 8233188 TI - Diabetes and the cost of coronary heart disease. PMID- 8233189 TI - Screening microtympanometry. PMID- 8233190 TI - Firearm related deaths in New Zealand 1978-87. AB - AIMS: To identify the role of firearms as a public health issue in New Zealand and to consider this information in the context of international research on potentially effective interventions for the prevention of firearm deaths. METHODS: National data on firearm related mortality for the period 1978 to 1987 were abstracted to identify all firearm deaths categorised as unintentional, suicide, homicide and of undetermined intent. RESULTS: Over this 10 year period, there were an average of 91 firearm deaths annually, accounting for 0.3% of all deaths. Suicides accounted for 75.5% of these deaths, unintentional deaths for 11.6% and homicides for 10.6%. No significant temporal changes were observed. CONCLUSIONS: While deaths from firearms are not a major public health problem in New Zealand, there is still a need to identify strategies likely to lead to reductions in the current levels of firearm deaths. Appropriately targeted legislative and educational strategies may be effective in reducing such deaths. PMID- 8233191 TI - New Zealand general practitioner referral patterns. AB - AIM: The aim of this study was to describe the referral patterns of general practitioners in New Zealand, for a defined list of medical conditions. METHODS: A postal questionnaire was sent to a random sample of 200 general practitioners, selected from the Medical Council of New Zealand list. Responses were made on five point scales to indicate general practitioners' views on the appropriateness of referral. For each medical condition, derived from An Educational Guide for General Practice, respondents also indicated favoured agencies for referral. Cluster analysis was used on an SPSSX dataset. RESULTS: A response rate of 87% (91.5% of eligible contacts) was achieved. The conditions for which referrals were least often made included mild hypertension, rheumatoid arthritis not requiring second line therapy, hypothyroidism, and obesity. Referral was common for rheumatoid arthritis requiring second line therapy, insulin dependent diabetes mellitus, altered bowel habit with rectal bleeding, multiple sclerosis, and postmenopausal bleeding. Seventy seven different referral agencies were indicated by respondents: the most common was the general physician (13% of all agencies indicated). CONCLUSIONS: Although there are a large number of conditions which are routinely managed only in general practice, general practitioners need to be able to refer patients to other health carers, even if only occasionally. There is a high degree of consensus as to the conditions for which referral is usually appropriate. They tend to be conditions requiring well defined investigation, treatment or management procedures which are commonly provided by centralised secondary or tertiary institutions. PMID- 8233192 TI - Prevention of cytomegalovirus disease in transplant recipients. PMID- 8233193 TI - Trends in primary medical care related services and expenditure in New Zealand 1983-93. AB - AIMS: To analyse recent trends in primary medical care (PMC) related expenditure, general medical services (GMS), pharmaceuticals, laboratory, Accident Compensation Corporation (ACC) and other categories, as a basis for general practitioners, other primary health care providers and regional health authorities becoming better positioned to make more effective decisions about the use of this expenditure. METHODS: Data from the Department of Health, Accident Compensation Corporation and other sources of this expenditure over the period 1983-93 were obtained and analysed. RESULTS: Primary medical care related expenditure between 1982-3 and 1990-1 increased annually at an inflation adjusted rate of 6.1% to a total of $1006.2M. In 1991-2 there was a slight inflation adjusted decrease in overall primary medical care related expenditure of 0.5% although maternity benefits increased by 20.7%. An overall increase of 2.7% occurred in 1992-3 with a 10.1% increase in laboratory benefits and 16.4% in maternity benefits. Price, volume, mix and other factors were analysed to explain these increases. The overriding determinants of the cost increases are the volume of primary medical care related services such as pharmaceuticals, laboratory and maternity services and the mix effect ie, more expensive drugs, tests, etc. CONCLUSION: Alternative strategies for control of primary medical care related service utilisation costs are discussed. It is suggested that the most effective strategy is some form of general practitioner or community based budget holding within which general practitioners and/or communities are empowered to make decisions about their priorities in achieving the best possible quality and quantity of primary health care. PMID- 8233194 TI - Prevalence of insulin self manipulation in young women with insulin dependent diabetes. AB - AIMS: 1. To determine the prevalence of insulin self manipulation for the purposes of weight control, in a population based study of young women with insulin dependent diabetes. 2. To compare the North Canterbury prevalence of insulin self manipulation with figures reported in overseas studies. METHOD: All women with insulin dependent diabetes aged 18 to 30 years inclusive and resident in North Canterbury, were invited to an interview to document their diabetes self care behaviour. A blood sample was taken at the time of interview for measurement of glycated haemoglobin. RESULTS: Ninety three percent of eligible women agreed to participate in the study. Twelve of the 69 participating women (17%) admitted to reducing their insulin dose at some time in the past, for the purposes of weight loss. This figure is significantly lower than the 37% (p = 0.047), reported from a British study, and the figure of 39% (p = 0.006), reported from the United States. The median glycated haemoglobin of the twelve subjects who reported manipulating their insulin was slightly higher than that of the remaining 57 subjects, but this difference did not reach statistical significance (79 mmol/mol haem versus 71 mmol/mol haem, p = 0.07). CONCLUSIONS: The self reported prevalence of insulin manipulation for the purposes of weight loss appears to be lower in North Canterbury, compared to overseas studies. The reason for this difference is unexplained. Health care professionals should nevertheless be aware of this method of weight reduction, as it has been associated with poor diabetes control, clinical eating disorders and possible long term detrimental effects on health. PMID- 8233195 TI - The effect of maternal smoking on admission to hospital in infancy. PMID- 8233196 TI - The prevalence of bad headaches including migraine in a multiethnic community. AB - AIM: Overall and ethnic specific prevalences of bad headache including migraine, for the New Zealand population, are unknown. A study was carried out in South Auckland to estimate prevalence and to explore ethnic differences in doctor attendance for the diagnosis and management of bad headaches. METHODS: Telephone interviews were administered to respondents selected by random digit dialing of households. RESULTS: 40.6% of the respondents suffered from bad headaches. 54.5% of these had the characteristics of bad headache with features symptomatic of migraine. Trends in the prevalence of bad headache with features symptomatic of common migraine, peaked between the ages of 30-49 years in both men and women. A difference was seen in the prevalence of bad headache with aura, with or without common migraine features, when ethnic groups and gender were examined. The difference in prevalence of aura was particularly noticeable between Pacific Island men and women. Although there was no difference between ethnic groups in doctor attendance, headaches were more likely to be labelled as migraine in Europeans than in the Polynesian groups. CONCLUSION: Ways in which people perceive and report their bad headaches have a bearing on management by general practitioners. Although no overall ethnic predominance was seen, there was a gender difference amongst Pacific Island people in reporting bad headaches with aura. The labelling process, and thus the management by general practitioners does demonstrate likely ethnic differences. PMID- 8233198 TI - New patient confidentially rules--good or bad. PMID- 8233197 TI - Alzheimer's disease and vegetable oils. PMID- 8233199 TI - Love sick. PMID- 8233200 TI - The Medical Council and transmission of major viral infections. PMID- 8233201 TI - Pouring gasoline on an incendiary issue. PMID- 8233202 TI - A camp health aide? PMID- 8233203 TI - Freedom of choice. PMID- 8233204 TI - Why nurses lose their licenses--Part II. PMID- 8233205 TI - AONE's healthcare reform transition. American Organization of Nurse Executives. PMID- 8233206 TI - Ethics and economic pressure: a case in point. PMID- 8233207 TI - Nurses in public policy. PMID- 8233208 TI - OBRA 1993 slices Medicare payments. Omnibus Budget Reconciliation Act. PMID- 8233209 TI - Beware the runaway IS. PMID- 8233210 TI - Maximizing limited resources through TEAMCARE. AB - A restructuring of staffing for a 30-bed orthopedic unit is described. A team nursing approach was initiated after careful delineation of the roles of the various nursing personnel. Specific training of non-RNs expanded their skill base and versatility. Productivity of non-licensed staff increased markedly and RN focus shifted from task completion to managing and directing care. PMID- 8233211 TI - A new documentation tool. AB - A Gwinnett Hospital System task force successfully establishes a 24-hour patient care record which consolidates paperwork and, when necessary, eliminates it to use nursing knowledge and time more efficiently. PMID- 8233212 TI - Understanding variances in hospital stay. AB - Systems for tracking, analyzing, and reporting variances in the length of hospital stay enable nurse case managers, nurse managers, and nurse executives to ensure desirable clinical outcomes in a cost-effective manner. PMID- 8233213 TI - Part II--clinical information systems: the nursing interface. AB - Given the fact that nurses generate over 50 percent of patient information, usable, automated nursing systems are essential. Neglecting the issues of usability of computer systems in healthcare results in poorly designed user interfaces, leaving nurses battling against extremely difficult and hostile systems. Two state-of-the-art technologies and nursing interfaces needed to develop mission-critical Clinical Information Systems are explored. PMID- 8233214 TI - Developing a nursing managed central transportation service. AB - Using computer simulation, a centralized transportation system was designed which resulted in 1) more time at the bedside for nurses and technicians, 2) fewer FTEs needed for transporting, 3) less waiting time for patients, and 4) better overall control of the transportation system. PMID- 8233215 TI - A psychiatric patient classification system that works! AB - Having used a Patient Classification System for six years which had limited psychiatric indicators and data processed elsewhere at considerable delay, Alberta Hospital Ponoka implemented a new system which has met with great success. Generic workload indicators were developed by staff. These could be modified easily to meet various unit needs. Relatively accurate prediction of staffing needs greatly facilitates budgeting. PMID- 8233216 TI - Health reform proposals: which is the fairest of them all? PMID- 8233217 TI - Moving out of the middle: transition to nurse executive. AB - Many factors should be considered by persons thinking about a move from midlevel to top-level nursing administration. A number of different programs exist, all of which involve mentoring with the chief nurse executive for a time. The seven month VA program is described, which trains for the positions of Associate Chief for Education, Supervisor, Nursing Home Care Unit and Assistant Chief Nurse. Key steps include 1) developing a clear understanding of the proposed role, 2) getting personal affairs in order and 3) considering the fiscal implications. PMID- 8233218 TI - Job sharing and the clinical nurse specialist role. AB - Shared employment is a promising form of flexible scheduling for career-oriented CNSs who wish to remain in the workforce but at the same time need to work part time temporarily to attend to family responsibilities. The organization benefits from increased productivity and less turnover. While job sharing is known to be extremely successful, documenting more wide-spread practice of this work option is necessary. Nurse administrators should be sensitive to the parental concerns of nurses, particularly CNSs, whose valuable knowledge and skills will continue to be in high demand in the healthcare marketplace. PMID- 8233219 TI - Implementing the Patient Self-Determination Act. AB - An interdisciplinary process action team developed policies and procedures for implementation of the Patient Self-Determination Act. Specific roles were defined for all involved in the process: Admissions Office, Unit Nurse, Social Worker, Attending Physician, Medical Records, Public Affairs, Patient Administration, Legal Office, Medical Library, Medical Law and Chaplain. An educational effort provides orientation for the hospital staff and the community on development of advance directives by patients. PMID- 8233220 TI - Life and death decisions: Mr. H. AB - Touched by a tragic situation involving one patient's quest for a peaceful, dignified death, a critical care nurse describes the scenario involved. A compassionate resolution can be reached by examining all medical aspects- psychosocial as well as physical. Decisions affect not only our patients and their families, but ultimately our profession. PMID- 8233221 TI - Developing a unit-based council in the medical ICU. AB - A unit-based council provided an environment within a medical ICU which increased commitment and improved morale. All healthcare members are encouraged to participate in the unit's decision-making process while working towards common goals--participative management and quality patient care. PMID- 8233222 TI - The nursing plan: innovative home health documentation. AB - In order to improve documentation, a home health agency task force analyzed and revised its current nursing care plan system. Nurses in the five-county service area were involved in all aspects of the development. The system combines nursing interventions and patient outcomes with the overall physician plan to generate a totally computerized treatment plan. PMID- 8233223 TI - House supervisors: the pulse of hospital life. PMID- 8233224 TI - "Focus" on patient care plans. PMID- 8233225 TI - The effect of normal infant maturation on nuclear medicine studies. PMID- 8233226 TI - Nuclear medicine and AIDS. AB - The human immunodeficiency virus (HIV) infection and its associated illnesses in a relatively young population of patients provides an expanding role for nuclear medicine. The disease enforces a review of each department's infection control procedures. It also necessitates a widening of the differential diagnosis in high risk patients. It has also resulted in an increase in the number of patients presenting with diseases such as Pneumocystis carinii pneumonia, Kaposi's sarcoma etc. which prior to the HIV epidemic were extremely rare. Thus in high risk patients the interpretation of abnormalities in nuclear medicine scans needs to include the spectrum of opportunistic infections and unusual tumours. The presence of opportunistic infections in the severely immunocompromised patient has lead to the development of techniques not normally used, i.e. lung 99Tcm diethylenetriamine pentaacetate (DTPA) transfer/clearance, donor leukocyte scanning to allow rapid diagnosis of an abnormality. Radionuclide techniques are also used to monitor the effect of therapy directed at the HIV itself or against opportunistic infections. This review covers aspects of infection control as well as the use of radionuclides to investigate specific problems related to HIV infection and therapy of the associated disease processes. PMID- 8233227 TI - The role of nuclear medicine in acute gastrointestinal bleeding. AB - In most patients with upper gastrointestinal (GI) bleeding endoscopy will locate the site and cause of bleeding, and also provide an opportunity for local therapy. The cause of lower GI bleeding is often difficult to attribute, even when pathology is found by colonoscopy or barium enema. Nuclear medicine techniques can be used to identify the site of bleeding in those patients in whom the initial diagnostic procedures are negative or inconclusive. Methods using transient labelling of blood (e.g. 99Tcm-sulphur colloid) produce a high target to-background ratio in positive cases, give quick results and localize bleeding sites accurately, but depend upon bleeding being active at the time of injection. Techniques using stable blood labelling (e.g. 99Tcm-labelled red blood cells) may be positive even with intermittent bleeding but may take several hours to produce a result and are less precise in localization. In order for these methods to become more widely accepted by physicians and surgeons, and for them to be cost effective, patients should be carefully selected. The most useful application is in patients with recurrent or prolonged bleeding, those with inconclusive endoscopy or barium studies, and those who are high-risk surgical candidates. PMID- 8233228 TI - Treatment of malignant phaeochromocytoma, paraganglioma and carcinoid tumours with 131I-metaiodobenzylguanidine. AB - 131I-metaiodobenzylguanidine (MIBG) has been used for the therapy of neural crest tumours. We report our experience with this agent in nine patients (malignant phaeochromocytoma 2; malignant paragangliomas 3; malignant carcinoid tumours 4). Six patients had soft tissue metastases alone while three had both soft tissue and bone metastases. The follow-up period ranged from 8 to 110 months, the number of doses ranged from one to seven with 3.1 to 11.1 GBq per administration and a cumulative activity from 4.8 to 40.1 GBq, while the tumour response was assessed according to World Health Organization (WHO) criteria. None of the patients showed complete tumour regression. Five patients showed complete symptomatic and three showed complete hormonal responses. Three patients with carcinoid tumours died. 131I-MIBG is thus a useful therapeutic modality which provides temporary palliation of these difficult tumours, but is rarely curative. PMID- 8233229 TI - Evaluation of the ratio method compared with graphical analyses for estimating nigrostriatal function in human 18F-dopa PET studies with or without carbidopa. AB - The striatal to cerebellar (S/C) activity ratio for estimating nigrostriatal function was compared with the Patlak analyses in 18F-dopa (FD) positron emission tomography (PET) with pretreatment with 100 mg carbidopa (CD). Two different time activity curves of plasma FD or cerebellar 18F were used for the Patlak analyses. The S/C ratio increased linearly with time for 120 min and the ratio at 120 min correlated closely with the uptake constants by the two Patlak analyses (r = 0.94, 0.96) in six normal volunteers and six parkinsonism patients. The S/C ratio and the uptake constant by the cerebellar Patlak analysis without CD also showed a fairly good correlation (n = 30, r = 0.85). Then, the S/C ratios with and without CD were compared. Since CD increased both the striatal and the cerebellar radioactivities proportionally within each subject, CD did not change the S/C ratios. The S/C ratios both with and without CD were simple and comparable to the uptake constants in the FD PET studies. PMID- 8233230 TI - The evaluation of gallbladder function by quantitative radionuclide cholescintigraphy in patients with noninsulin-dependent diabetes mellitus. AB - Twenty-nine patients (24 males, 5 females; aged 62.8 +/- 10.1 years) with noninsulin-dependent diabetes mellitus (NIDDM) without gallstones and impaired liver function were included in our study. The patients were categorized by blood sugar control and disease duration. Twelve normal controls (10 males, 2 females; aged 62.8 +/- 12.4 years) were also studied for comparison. The gallbladder filling fraction (FF) and ejection fraction (EF) were calculated following the method of Krishnamurthy et al. The results showed no significant difference (P > 0.5) between (1) NIDDM and normal controls in FF (69.1 +/- 29.4 versus 67.5 +/- 24.0%) and EF (45.0 +/- 23.5 versus 54.8 +/- 10.0%) by a t-test, (2) good and poor blood sugar control in FF (64.9 +/- 30.9 versus 73.7 +/- 26.4%) and EF (42.0 +/- 25.4 versus 54.8 +/- 10.0%) by a Mann-Whitney U test, (3) long and short disease durations in FF (70.9 +/- 29.7 versus 67.9 +/- 28.7%) and EF (37.9 +/- 20.7 versus 50.0 +/- 24.1%) by a Mann-Whitney U test. In conclusion, the results challenge some previous reports and warrant further research. PMID- 8233231 TI - Imaging of renal cell cancer with radiolabelled octreotide. AB - Recently the presence of somatostatin receptors on human renal cell carcinomas has been demonstrated by autoradiographic techniques on surgically removed kidneys. In a prospective study we evaluated, by means of 111In-labelled octreotide scintigraphy, the in vivo tumour imaging in a group of patients with biopsy proven renal cell carcinomas at different tumour stages. Seven patients were studied. In three of them (43%) pathological tracer accumulation was demonstrated. In these patients 20 out of 23 known tumour localizations were clearly visualized. Tracer uptake could be inhibited by prior administration of cold octreotide. We conclude that 111In-octreotide scintigraphy can be used to demonstrate, in vivo, metastatic renal cell carcinoma. PMID- 8233232 TI - 99Tcm-labelled polyclonal human immunoglobulin G scintigraphy before and after intra-articular knee injection of triamcinolone hexacetonide in patients with rheumatoid arthritis. AB - The ability of 99Tcm-labelled polyclonal human immunoglobulin G (99Tcm-IgG) scintigraphy to monitor intra-individual variation in arthritis activity was studied in seven patients with rheumatoid arthritis (RA). These patients were treated with an intra-articular injection of 20 mg triamcinolone hexacetonide. The results of semiquantitative 99Tcm-IgG scintigraphy were compared with the degree of joint swelling and the histological changes observed in synovial biopsies before and 14 days after the injection. In all seven patients the local treatment resulted in a decreased arthritis activity of the treated knee as measured clinically or histologically. This decrease was parallelled, in all patients except one, by a lower uptake of 99Tcm-IgG after the injection when compared to uptake prior to treatment. This study shows that 99Tcm-IgG scintigraphy is able to reflect intra-individual variations in arthritis activity in patients with RA. PMID- 8233233 TI - A comparative study of labelled neutrophil behaviour in patients with rheumatoid arthritis and normal volunteers. AB - A comparative study of neutrophil pharmacokinetics and distribution between normal volunteers and patients with rheumatoid arthritis (RA) is presented. White cell viability was confirmed by prominent localization of activity in the spleen with minimal liver uptake and lack of retention in the lungs in both normal volunteers and RA patients. There were no significant differences between both groups in granulocyte clearance from the lungs and their subsequent distribution in the spleen, bone marrow and liver. The results of this study demonstrate that the behaviour of granulocytes in RA patients is essentially unchanged from that of normal volunteers. PMID- 8233234 TI - Abstracts of the Fifth European Symposium on Radiopharmacy and Radiopharmaceuticals. Cambridge, U.K., 21-24 March 1993. PMID- 8233236 TI - Question of the month--May 1993. PMID- 8233235 TI - Cerebral perfusion deficits in divers. PMID- 8233237 TI - Sacroiliac quantitative bone scintigraphy in ankylosing spondylitis: any clinical relevance? PMID- 8233238 TI - New perspectives in radioimmunoassay. AB - In addition to providing the basis of 'ultrasensitive' immunoassays, nonisotopic labels permit development of 'user friendly' methodologies, including simple devices designed for extralaboratory use and intended to provide yes/no answers to basic diagnostic questions, for example, the presence of an infectious disease, the commencement of pregnancy etc. Immunosensors are also intended to fulfil these objectives (albeit yielding quantitative information), but despite much research the prospect of adequately sensitive instruments of this type remains remote. Finally, nonisotopic labels enable the development of a new generation of miniaturized multianalyte assays permitting in principle the determination of entire analyte menus in small samples of body fluids. Such technologies will resemble those employed in compact disk recorders and other data-storage devices, differing only in that light emitted from discrete zones on a solid surface is fluorescent (or possibly chemiluminescent) rather than reflected, and yields chemical rather than physical information. The emergence of such technologies, will, inter alia, facilitate the screening of entire populations for infectious disease, endocrine dysfunctions, genetic abnormalities etc. using, for example, finger-tip blood samples, throat and endocervical swabs, and hopefully will contribute to the large-scale diagnosis and ultimate reduction of disease. PMID- 8233239 TI - Radionuclides and therapy of thyroid cancer. AB - The majority of thyroid carcinomas are removed surgically. The appropriate surgical technique is still debated. After surgery the amount of residual thyroid or tumour and the presence of local or distant metastases is often in doubt, particularly if it is not detectable clinically. Therefore, methods for determining the presence of disease or the later recurrence of disease are needed. They commonly include serum thyroglobulin and imaging after diagnostic or therapeutic doses of 131I. Other techniques are used such as 131I whole body retention (using a whole body counter), 201Tl and 99Tcm-sestamibi imaging. The place of these diagnostic methods in the management of thyroid cancer is reviewed in this article. Radioiodine would seem an ideal treatment for recurrence of functioning thyroid carcinoma as 131I targets the lesion and has minimal side effects. However, the indolent nature of well-differentiated thyroid carcinomas makes it difficult to assess the benefits of radioiodine therapy both in its ability to ablate the normal thyroid and to treat recurrent and metastatic disease. However, the addition of radioiodine therapy to local surgical removal reduces both the occurrence of metastases and the morbidity with prolonged follow up. Unresolved issues that remain concern the activities of radioiodine needed to achieve adequate ablation of residual thyroid tissue and to treat residual and recurrent cancer. There is also debate as to exactly which patients require radioiodine therapy. This review also considers radiation protection and the side effects of 131I therapy. PMID- 8233240 TI - Analysis of radiation doses received by the public from 131I treatment of thyrotoxic outpatients. AB - Radiation doses to the family and general public from 131I therapy of outpatients were assessed for comparison with the 1990 International Commission on Radiation Protection (ICRP) dose limits. Doses to family members of such patients, measured by previous workers, were converted to show that on average 5.6 +/- 3.7 microSv MBq-1 is received by the spouse and 1.5 +/- 0.87 microSv MBq-1 by other family members. It was deduced that the average dose to the spouse (D microSv MBq-1) when couples sleep apart for T days after administration is given by: D = 2.528 + 3.072e-0.11T if an effective half-life, T(eff), of 6.35 days is used. The cumulative effect of repeated treatments should be considered and in view of ALARA it is recommended that couples sleep apart for at least 14 days after each administration, even if this is below limits permitted by authorities. The dose to a pregnant colleague or family member other than the wife should be below 2 mSv if the total administered activity is not more than 600 MBq. If the wife is pregnant, couples should sleep apart for at least 30 days after administration. PMID- 8233241 TI - Cellular versus vascular rejection in transplant kidneys. Correlation of radionuclide and Doppler studies with histology. AB - The presence of two distinct subtypes of renal allograft rejection are well documented by histological studies. The differentiation between vascular rejection (VR) and cellular rejection (CR) is essential for proper management by avoiding the need for unnecessary and potentially harmful immunosuppressive treatment of VR. A histological pattern with features that are similar and confusable with some cases of rejection may be seen in cyclosporin A toxicity (CyT). To evaluate the efficiency of Guy's perfusion index (GPI) and the Doppler pulsatility index (DPI) in differentiating these two histological subtypes, a prospective study was designed in which a total of 140 radionuclide tests and 133 ultrasounds scans performed on the same day on 58 patients during the first 3 months post-transplant were analysed, and the results correlated with the histological findings of 84 renal biopsies. Results show that the GPI had a sensitivity of 86.5% and a specificity of 94% in differentiating VR and CyT from CR, while the DPI had values of 83% and 69%, respectively. Chi-squared analysis showed a higher significant association between the GPI and histology (P < 0.0001) compared to that of the DPI and histology (P < 0.005), while Youden's index (J) showed a significant difference (P < 0.05) between GPI and DPI. It is concluded that GPI is more sensitive and specific than DPI in differentiating transplants that are well perfused from those with poor perfusion (VR and CyT). PMID- 8233242 TI - Detection of lung and bone infection with anti-granulocyte monoclonal antibody BW 250/183 radiolabelled with 99Tcm. AB - Twenty-four patients with suspected infection (eight bone, 16 lung) were studied using monoclonal antibody BW250/183 which recognizes epitopes present on the surface of granulocytes. Bronchofibroscopic samples (microbiological studies and alveolar cell counts) were obtained from 14/16 patients with lung disease. Bronchofibroscopy isolated a micro-organism nine times. In two other cases, the diagnosis of infection was based on clinical course data. Infection was confirmed by surgical biopsy in the eight patients with bone pathology. Scans were performed 2 and 24 h after injection of 1 mg BW 250/183 labelled with 99Tcm. For lung disease, immunoscintigraphy was positive six times (five true positive, one false positive) and negative 10 times (six false negative, four true negative). Immunoscintigraphy was false negative when the lung infection was not systematized or no granulocytes were mobilized in the infectious site. Immunoscintigraphy was falsely positive when noninfectious lung disease mobilized granulocytes. It was positive in all patients with bone infection. Images recorded at 24 h had better sensitivity (five false negative) than those at 2 h (eight false negative). PMID- 8233243 TI - Radioimmunoscintigraphy of colorectal carcinoma with a 111In-labelled anti-TAG-72 monoclonal antibody. AB - Radioimmunoscintigraphy (RIS) of colorectal carcinoma with a 111In-labelled anti TAG-72 monoclonal antibody (CYT-103) has been performed in 24 patients with five primary lesions and 10 suspicious of recurrence (in one of these patients two RIS were made). Histopathological confirmation of the disease and surgical liver examination were carried out in all primary tumours and in 12 possible recurrences. In the remaining eight patients a final diagnosis was established according to the clinical course and other diagnostic procedures. Planar and tomographic scans were obtained at 48 and 72 h postinjection in all patients. All primary tumours were detected by RIS. In the group with recurrences confirmed pathologically the results were nine true positive, two true negative and one false positive in a patient affected only with liver disease. The nine true positive studies corresponded to four positive by computed tomography (CT), four negative by CT and one nonconclusive by CT. Surgical liver examination results were 15 true negative and two false negative. No correlation was found between serum levels of carcinoembryonic antigen or TAG-72 and the detection of the lesions. In 10 patients human anti-mouse antibody (HAMA) levels were studied. In conclusion, RIS with an anti-TAG-72 monoclonal antibody is a useful technique for the study and localization of colorectal tumours, mainly in cases of recurrence, being also indicated in patients with normal TAG-72 serum levels. PMID- 8233244 TI - Radioimmunoscintigraphy of recurrent colorectal carcinoma using 111In-labelled murine monoclonal antibody B72.3: a comparison with contrast-enhanced computed tomography. AB - Ten patients underwent contrast-enhanced abdominal computed tomographic (CT) carcinoembryonic antigen (CEA) measurement and murine 111In-B72.3 monoclonal antibody radioimmunoscintigraphy for recurrent or residual colorectal carcinoma. All patients had undergone primary colorectal tumour resection at a median of 3 (range 1-18) months previously. Histological analysis of pelvic tissue biopsies confirmed that six patients had recurrent or residual colorectal cancer and that four were tumour free. Whilst scintigraphy correctly showed all six patients with recurrent or residual disease, both CT and CEA measurement were correct in only four patients. In the four patients without recurrent cancer, CT was correct in two patients whilst scintigraphy and CEA measurement were accurate in three patients. Six out of the 10 patients showed marked nonspecific colonic excretion of radiolabelled antibody which was correctly interpreted in five patients. Future prospective studies comparing CT scans, CEA estimation and B72.3 radioimmunoscintigraphy in a larger group of patients with suspected residual or recurrent colorectal adenocarcinoma may help to define the respective sensitivity and specificity of these techniques. PMID- 8233245 TI - Sensitivity and specificity of 99Tcm-HMPAO single-headed SPECT in dementia. AB - The sensitivity and specificity of 99Tcm-hexamethylpropyleneamine oxime (HMPAO) single-headed single photon emission computed tomography (SPECT) in dementia were evaluated in elderly patients with a mean age of 84 years suffering from dementia (n = 59) and compared to an age- and sex-matched control group with normal cognitive function (n = 14). The demented patients were classified as suffering from primary degenerative dementia (PDD, n = 51) based on DSM-III-R criteria. Moreover, PDD patients were divided into three subgroups of severity of cognitive impairment, according to their Folstein score. Multi-infarct dementia (MID, n = 8) was diagnosed based on clinical features, computed tomographic findings and Hachinski ischaemic score. Images were assessed qualitatively by visual interpretation of shades of colour in cortical regions. A SPECT defect confined to the frontal, parietal, temporal or (parieto) occipital lobe was defined as one lesion. The mean number of SPECT lesions was 1.4 (range 0-4) in the control group, 2.4 (range 0-8) in the PDD group and 2.9 (range 2-5) in the MID group and showed a significant difference (P < 0.02) between groups. To obtain an acceptable level of specificity of 64.3%, a cut-off value of three lesions had to be chosen. In the PDD group, sensitivity was then 25% for the mildly, 43.8% for the moderately and 46.7% for the severely affected PDD subgroup. In the MID group sensitivity was then 75%. Interestingly, in this elderly patient population the location of lesions was homogenously distributed in all groups, including the control group. PMID- 8233246 TI - Scintigraphic visualization of acute pancreatitis in cats with 99Tcm-citrate. AB - The accumulation of 99Tcm-citrate in abscesses and arthritic joints has previously been demonstrated. In the present study its usefulness was evaluated in 10 mongrel cats with acute pancreatitis induced by metronidazole infusion. Five cats, used as normal controls, were given physiological saline. All 15 cats were injected intravenously with 185 MBq 99Tcm-citrate. Scintigrams were obtained at 30 min intervals up to 2 h after injection by a gamma camera. The cats were then sacrificed. Tissue samples were obtained, weighed and counted against a 1/1000 dilution of the injected dose. In normal cats the kidneys and the urinary bladder were the only organs that could be visualized. In acute pancreatitis the pancreas and its exudate in the abdomen were also evident on the images. The pancreas to other neighbouring organ ratios increased 2.3-9.7 fold in acute pancreatitis compared to normal controls. Gross autoradiograms showed localization of radioactivity in the lobules of pancreas. Histologic examination of sections of pancreas demonstrated oedema, haemorrhage and some fat necrosis in pancreatic parenchyma, confirming acute pancreatitis. In conclusion, it has been demonstrated that acute pancreatitis in cats can be visualized scintigraphically with 99Tcm-citrate. PMID- 8233247 TI - Regional distribution of 201Tl during one-leg exercise: comparison with leg blood flow by plethysmography. AB - To validate the use of 201Tl distribution as an estimate of regional blood flow in the legs, 201Tl leg uptake was compared by whole body scintigraphy and simultaneously measured leg blood flow by plethysmography during one-leg exercise in 11 male subjects. 201Tl leg uptake ratio and leg blood flow ratio were also compared to exclude the effect of cardiac output variation in each subject. There was a good correlation between 201Tl leg uptake and leg blood flow (r = 0.85, P < 0.01, y = 0.28x + 1.00). Moreover, there was a highly linear correlation between these ratios (r = 0.98, P < 0.01, y = 0.75x + 0.13), although the 201Tl leg uptake ratio somewhat underestimated the leg blood flow ratio as exercise became strenuous. It is concluded that 201Tl leg uptake reflects regional blood flow in the legs during exercise. PMID- 8233248 TI - Scintigraphic evaluation of mycetoma. AB - Mycetoma is a rare deep mycosis caused by certain fungi. Clinical findings include enlargement and tumefaction of the affected region accompanied by grains coming out of the lesions. The feet are the most frequently affected area. Four patients have been studied who presented with mycetoma of the feet. Gallium scintigraphy was always abnormal in the affected areas and bone scintigraphy was abnormal in three of the four patients. The patient who had a normal bone study had received effective treatment (7 years). In the other patients the results were poor. Scintigraphic findings seem to be similar to those noted in cases of acute osteomyelitis associated with cellulitis, despite long-term therapy. Bone and gallium studies may be of possible value for therapy follow-up in cases of mycetoma. PMID- 8233249 TI - Radiochemical stability during infusion of 131I-labelled metaiodobenzylguanidine for therapeutic use. AB - 131I-labelled metaiodobenzylguanidine ([131I]MIBG) is used both for diagnostic scintigraphy and for radionuclide therapy of neural crest derived tumours in particular neuroblastoma, malignant phaeochromocytoma and paraganglioma. This paper presents data on radiochemical stability during the infusion of 3.8, 5.7 and 7.7 GBq [131I]MIBG in 100 ml infusion fluids for therapy. The period of investigation started at the moment of dilution of the thawed infusion concentrates (t = 0), which arrived in a frozen condition from the manufacturer, and ended at the termination of infusion (t = 7 h). In 7 h the percentage of free [131I]iodide increased from 3.74 to 5.82, from 3.52 to 6.02 and from 3.72 to 6.40% in the 3.8, 5.7 and 7.7 GBq [131I]MIBG infusion fluids, respectively. The 0 7 h increases of the different radioactive concentrations did not differ significantly. All the infusion fluids contained less than 7% free [131I]iodide at the end of infusion (t = 7 h). PMID- 8233250 TI - A comparison of the accuracy of the TDx-FLM assay, lecithin-sphingomyelin ratio, and phosphatidylglycerol in the prediction of neonatal respiratory distress syndrome. AB - OBJECTIVE: To ascertain whether the TDx-FLM assay is a better predictor of neonatal respiratory distress syndrome (RDS) than the lecithin-sphingomyelin ratio (L/S) and phosphatidylglycerol (PG). METHODS: Amniotic fluid samples of 140 women requiring assessment of fetal lung maturity and delivering within 72 hours of amniocentesis were analyzed by the three methods. The accuracy in predicting RDS was assessed by conventional statistical techniques. RESULTS: The TDx-FLM test had a better sensitivity (89.6 versus 48.2%; relative risk [RR] 3.9, 95% confidence interval [CI] 1.35-11.23, P = .001) and negative predictive value (96.4 versus 86.8%; RR 0.36, 95% CI 0.12-1.04, P = .02) than the L/S. However, the L/S had a better specificity (89.1 versus 73.8%; RR 0.64, 95% CI 0.49-0.82, P = .005) than the TDx-FLM assay. The PG test had a predictive ability similar to that of the TDx-FLM assay. However, there was no significant difference in the overall accuracy of the three tests, and the areas under the receiver-operating characteristic curves for the L/S and TDx-FLM (0.80 and 0.87) were similar. CONCLUSIONS: The TDx-FLM assay performs similarly to the L/S and PG tests in the prediction of RDS and is quantitative, rapid, and reproducible. PMID- 8233251 TI - A randomized comparison of 15- and 40-minute dosing protocols for labor augmentation and induction. AB - OBJECTIVE: To compare two low-dose oxytocin protocols in terms of fetal distress, uterine hyperstimulation, cesarean delivery rate, maximum dose of oxytocin, and length of labor. METHODS: We randomized 865 patients into 15-minute (incremental dose 1 mU/minute until 5 mU/minute, then 1 or 2 mU/minute) or 40-minute (incremental dose 1.5 mU/minute until 7 mU/minute, then 1.5 or 3.0 mU/minute) low dose protocols. Before oxytocin use, all subjects were stratified according to parity and purpose of oxytocin, ie, for induction or augmentation of labor. RESULTS: The 40-minute dosing protocol had a significantly lower maximum dose of oxytocin (augmentation, 6.5 versus 8.2 mU/minute, P < .001; induction, 11.5 versus 14.5 mU/minute, P < .001), a lower incidence of uterine hyperstimulation (augmentation, 18.8 versus 31.8%, P < .001; induction, 19.1 versus 33.0%, P < .002), and less fetal distress (augmentation, 15.5 versus 26.1%, P < .005). No significant differences were found in the cesarean rate or length of labor. CONCLUSION: A dosing interval of 40 minutes led to lower incidences of uterine hyperstimulation and fetal distress, and decreased the maximum dose of oxytocin, without affecting the length of labor or the cesarean rate. PMID- 8233252 TI - Prostaglandin E2-induced ripening of the human cervix involves changes in proteoglycan metabolism. AB - OBJECTIVE: To elucidate how prostaglandin E2 (PGE2) induces cervical ripening. METHODS: Cervical biopsies were obtained immediately postpartum from women successfully treated with PGE2 gel intracervically. Six specimens were incubated with [35S]sulfate and five were used to characterize the nonlabeled proteoglycan composition. In separate experiments, biopsy specimens from three term pregnant women with unripe cervices were incubated with PGE2 in organ cultures. Proteoglycans were isolated and characterized using ion-exchange and gel chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. RESULTS: During PGE2-induced cervical ripening, the synthesis of proteoglycans, especially a large chondroitin/dermatan sulfate proteoglycan and biglycan, increased three- to sixfold. This resulted in a net increase in the large proteoglycan in the PGE2-treated cervices. In organ culture, on the contrary, incubation with PGE2 decreased the proteoglycan synthesis. CONCLUSION: Prostaglandin E2-induced cervical ripening is accomplished by increased remodeling of the cervical connective tissue, involving changed proteoglycan metabolism and composition. PMID- 8233253 TI - A review of postmenopausal hormone therapy recommendations: potential for selection bias. AB - OBJECTIVE: To survey the advice given to physicians in regard to postmenopausal hormone therapy, and thus to find out indirectly whether selective prescribing may have contributed to the findings of health effects. DATA SOURCES: Surveys on the health benefits of postmenopausal therapy were identified from previous reviews. Experts in various fields were consulted regarding relevant textbooks, and recommended booklists were used. METHODS OF STUDY SELECTION: Most of the surveys were conducted in the United States and concerned only estrogens; therefore, we focused on these data. Advice given to United States physicians on estrogen prescribing was abstracted from the Physicians' Desk Reference, five textbooks, and other sources written between 1945-1990. DATA EXTRACTION AND SYNTHESIS: In advice given to physicians, there was concern that estrogens could cause cancer. In addition, it was not recommended to give estrogens to women in poor general health. Advice concerning cardiovascular diseases and estrogens was contradictory and variable. CONCLUSIONS: If the advice observed reflects the prescribing practices of physicians, then surveys on the health impact of estrogen therapy may have underestimated the risk of breast cancer and overestimated the prevention of fractures. The impact on studies of cardiovascular diseases is less clear, and other mechanisms of selection bias may have been more important. PMID- 8233254 TI - An ethical justification for emergency, coerced cesarean delivery. AB - OBJECTIVE: To provide an ethical justification for emergency coerced cesarean delivery without a court order when a pregnant woman refuses cesarean delivery. METHODS: Ethical analysis is conducted of the existing literature and an ethical justification illustrated by a clinical example. RESULTS: Absolute and near absolute objections to coerced cesarean delivery fail for lack of adequate arguments to show that the woman's autonomy is not constrained by obligations to the at-term fetal patient. The ethical justification for emergency coerced cesarean delivery requires that three criteria be satisfied: 1) high reliability of the prognostic judgment that on balance cesarean delivery is expected to prevent serious infant morbidity or mortality, 2) lack of physical resistance that could significantly increase the risks of maternal or fetal harm from coerced cesarean delivery, and 3) insufficient time to consider a court order. CONCLUSION: The obstetrician may justifiably coerce emergency cesarean delivery without a court order only when these three criteria are satisfied on a case-by case basis. PMID- 8233255 TI - Asthma in pregnancy. National Asthma Education Program Working Group on Asthma and Pregnancy. National Institutes of Health, National Heart, Lung and Blood Institute. AB - This manuscript is a summary of a comprehensive report dealing with asthma and pregnancy issued by the working group on Asthma and Pregnancy, National Institutes of Health (NIH), National Heart, Lung, and Blood Institute. The report was developed by a panel of obstetricians, pharmacologists, internists, allergists, and pulmonologists, who met over an 18-month period under the auspices of the NIH. Undertreatment of pregnant asthmatics, partially because of unfounded fears of adverse pharmacologic effects on the developing fetus, remains the major problem in the management of asthma during pregnancy in the United States. The four key components of asthma management during pregnancy are: 1) objective assessment of maternal lung function and fetal well-being, 2) avoidance or control of environmental precipitating factors, 3) pharmacologic therapy, and 4) patient education. PMID- 8233256 TI - Morphologic precursors of ovarian epithelial tumors. PMID- 8233257 TI - Ovarian management during radical hysterectomy in the premenopausal patient. PMID- 8233258 TI - Does midline episiotomy increase the risk of third- and fourth-degree lacerations in operative vaginal deliveries? PMID- 8233259 TI - Magnetic resonance imaging of pelvic floor relaxation: dynamic analysis and evaluation of patients before and after surgical repair. AB - OBJECTIVE: To evaluate structures involved in pelvic support using conventional and snapshot magnetic resonance imaging (MRI). METHODS: We used conventional spin echo MRI and dynamic snapshot GRASS MRI at various levels of the Valsalva maneuver to describe and quantitate the anatomy of pelvic floor relaxation and to assess anatomical changes produced by surgical repair. Ten female volunteers were evaluated to define normal anatomy and reference measurements. Five women with pelvic floor relaxation were evaluated before and after surgical repair. RESULTS: Static and dynamic MRI were more sensitive than clinical pelvic examination in assessing and grading pelvic floor relaxation. Quantitative results showed widening of the levator hiatus and more vertical lie of the levator plate postoperatively. Descent of the pelvic organs on maximal straining postoperatively was the same as that in normal volunteers. The posterior urethrovesical angle on MRI was more than 110 degrees in 14 of 15 continent subjects. CONCLUSIONS: Magnetic resonance imaging may be valuable in analyzing and assessing pelvic floor relaxation and in understanding anatomical changes occurring before and after surgical repair. The increased sensitivity of MRI in grading prolapse may make it useful in evaluating women with symptoms of pelvic floor relaxation but who have negative findings on clinical examination. PMID- 8233260 TI - The Q-tip test and urethrovesical junction mobility. AB - OBJECTIVE: To determine the accuracy of the Q-tip test to measure urethrovesical junction mobility. METHODS: One hundred fourteen women with urinary incontinence or genital prolapse were evaluated with a Q-tip test and perineal ultrasound scanning to measure urethrovesical junction mobility with strain. The sensitivity, specificity, and positive and negative predictive values of the Q tip test to diagnose urethrovesical junction hypermobility were calculated using ultrasound as the standard. Repeat Q-tip and ultrasound measurements were obtained in ten subjects and analyzed for reproducibility. RESULTS: The Q-tip test had a sensitivity of 25%, specificity of 78%, positive predictive value of 67%, and negative predictive value of 37%. The Q-tip and ultrasound measurements were reproducible, as the within-subject variation was significantly small when compared to between-subject variation (P = .0001). CONCLUSION: The Q-tip test is inaccurate for measurement of urethrovesical junction mobility and should not be used in the manner described to diagnose urethrovesical junction hypermobility. PMID- 8233261 TI - Morbidity associated with abdominal myomectomy. AB - OBJECTIVE: To review the recent experience at Yale-New Haven Hospital with abdominal myomectomy and to assess morbidity associated with the procedure. METHODS: The hospital records of 128 women who underwent abdominal myomectomies during a 39-month period were reviewed retrospectively. Particular attention was given to blood transfusion requirement and the febrile morbidity rate. RESULTS: The average (+/- standard error of the mean) intraoperative estimated blood loss was 342 +/- 37 mL. Five cases (4%) had an estimated blood loss greater than 1000 mL. Intraoperative or postoperative transfusion occurred in 26 cases (20%); 70% of the transfused patients received autologous blood only. The operation was converted intraoperatively to a hysterectomy in one case. There were no other intraoperative complications. Febrile morbidity occurred in 15 patients (12%). Three women (2%) experienced postoperative complications including paralytic ileus, wound infection, and deep venous thrombosis. CONCLUSION: Febrile morbidity and blood transfusions associated with abdominal myomectomy appear to occur at an acceptably low rate, especially when unnecessary blood transfusions are excluded. These findings should modify the way patients are counseled before abdominal myomectomy. PMID- 8233262 TI - The concentrations of collagen-associated amino acids are higher in GnRH agonist treated uterine myomas. AB - OBJECTIVE: To test the hypothesis that the effects of estrogen reduction on uterine leiomyoma regression are mediated through changes in cell density or the extracellular matrix. METHODS: Uterine myomas were obtained from 20 women who had received randomly either the GnRH agonist leuprolide acetate depot for 3 months or placebo. The biochemical and morphologic characteristics studied included: total protein, DNA, and amino acid concentrations; histologic appearance; collagen content; and nuclear density. RESULTS: The absolute and relative concentrations of hydroxylysine, hydroxyproline, glycine, and proline were significantly greater (P < .05) in uterine myomas from patients pretreated with a GnRH agonist compared with placebo-treated controls. The GnRH agonist was also associated with trends toward increased mean total protein, DNA, and nuclear density, but the differences did not reach statistical significance. CONCLUSIONS: The concentrations of the amino acids contained in collagen were significantly greater in uterine myomas from patients treated with the GnRH agonist compared to myomas from placebo-treated controls. In addition, our observations suggest that the reduction in uterine myoma volume associated with GnRH agonist therapy is associated with alterations in the extracellular matrix. PMID- 8233263 TI - Randomized controlled study of premenstrual symptoms treated with ear, hand, and foot reflexology. AB - OBJECTIVE: To determine whether reflexology therapy--the application of manual pressure to reflex points on the ears, hands, and feet that somatotopically correspond to specific areas of the body--can significantly reduce premenstrual symptoms compared to placebo treatment. METHODS: Thirty-five women who complained of previous distress with premenstrual syndrome (PMS) were randomly assigned to be treated by ear, hand, and foot reflexology or to receive placebo reflexology. All subjects completed a daily diary, which monitored 38 premenstrual symptoms on a four-point scale. Somatic and psychological indicators of premenstrual distress were recorded each day for 2 months before treatment, for 2 months during reflexology, and for 2 months afterward. The reflexology sessions for both groups were provided by a trained reflexology therapist once a week for 8 weeks, and lasted 30 minutes each. RESULTS: Analysis of variance for repeated measures demonstrated a significantly greater decrease in premenstrual symptoms for the women given true reflexology treatment than for the women in the placebo group. CONCLUSION: These clinical findings support the use of ear, hand, and foot reflexology for the treatment of PMS. PMID- 8233264 TI - Effects of power and electrical current density variations in an in vitro endometrial ablation model. AB - OBJECTIVE: To investigate the thermal effects of electrical power and waveform on uterine tissue in vitro, using the gynecologic resectoscope with a 2.5-mm roller bar electrode. METHODS: The power setting was increased in a serial fashion using both the modulated or damped (coagulating) and unmodulated or undamped (cutting) current to ablate endometrial uterine tissue, using a technique similar to that employed in clinical situations. The power setting ranged from 20-75 W with the modulated waveform and 40-160 W with the unmodulated waveform. Measurements of tissue damage were made by staining for hematoxylin and eosin and the respiratory enzyme dehydronicotinamide adenine dinucleotide phosphate diaphorase. RESULTS: The amount of thermal injury present correlated linearly with the amount of wattage used for either current. Regardless of the power used, the maximum amount of injury into the myometrium was 4.2 mm, representing 19% of the uterine wall thickness, well within the limits of safety. Destruction to 3 mm of myometrium was achieved significantly more often using an unmodulated waveform at greater than 90 W (chi 2, P = .03). CONCLUSIONS: Although the correlation between power and tissue damage achieved statistical significance, the relationship was weak; less than 12% of the observed variation was attributable to increasing wattage. Power and waveform alone do not appear to be clinically significant determinants of the amount of thermal injury occurring during endometrial ablation. PMID- 8233265 TI - A comparison of esterified estrogens with and without methyltestosterone: effects on endometrial histology and serum lipoproteins in postmenopausal women. AB - OBJECTIVE: To compare the efficacy and safety of esterified estrogens with and without methyltestosterone. METHODS: Twenty-six women participated in a double blind randomized trial for 6 months. Outcome measures included serum total and lipoprotein-bound cholesterol, vasomotor symptoms, vaginal cytology and endometrial histology, and chemistry values. Analysis of variance and t test statistics were used to assess differences. RESULTS: After 6 months of therapy, the treatment groups were comparable with regard to symptom scores, vaginal cytology and endometrial histology scores, and clinical laboratory test values. Treatment with esterified estrogens plus methyltestosterone significantly decreased total cholesterol, high-density lipoprotein cholesterol (HDL), HDL2, HDL3, and apolipoprotein A1 compared to esterified estrogens alone. CONCLUSIONS: Esterified estrogens with or without methyltestosterone were effective at reducing menopausal symptoms and were well tolerated over 6 months of continuous treatment. A significant decrease in cholesterol and apolipoproteins in the estrogen plus methyltestosterone group suggests a potentially adverse impact on the beneficial effect normally imparted by estrogen therapy. PMID- 8233266 TI - Evaluation of routine diabetes and lipid screening after age 35 in candidates for or current users of oral contraceptives. AB - OBJECTIVE: To evaluate the efficacy of a diabetes and lipid screening program in women aged 35 or older who either were currently using a low-estrogen-dose oral contraceptive (OC) (0.03-0.04 ethinyl estradiol) or who desired to initiate low dose OC therapy after using a non-OC method or following a recent pregnancy. METHODS: Healthy women with no history of abnormal glucose tolerance, hypertension, or hyperlipidemia aged 35 years and older were routinely screened for diabetes (fasting and 2-hour values after 75 g glucose) and dyslipidemia (fasting total cholesterol, high-density lipoprotein [HDL] and low-density lipoprotein [LDL] cholesterol, and triglycerides). Three groups of women were studied: women currently using low-dose OCs (OC group), women desiring to change to OC therapy (non-OC group), and women within 3 months post-pregnancy (postpartum group). RESULTS: The three groups were similar in age, parity, body mass index (BMI), and mean arterial pressure. The mean months of contraceptive use were not different between the OC group (30.2 months; 95% confidence interval [CI] 26.6-33.8) and the non-OC group (38.4 months; 95% CI 32.0-44.8). The mean fasting serum glucose, after adjusting for age and BMI, was significantly lower in the OC group (82.4 +/- 9.4 mg/dL) than in the non-OC and postpartum groups (87.2 +/- 12.5 and 86.8 +/- 13.0 mg/dL, respectively; P = .01). After adjustments, no differences between the groups were found in the 2-hour post glucose levels, triglycerides, or HDL or LDL cholesterol levels. The non-OC group was found to have a significantly lower total cholesterol level (188.7 +/- 32.4 mg/dL) than the OC and postpartum groups (202.7 +/- 38.1 and 204.4 +/- 41.0 mg/dL; P = .02) after adjusting for age and BMI. This association lost its significance after applying multiple range tests, maintaining an alpha error of 0.05. CONCLUSION: The absence of significant adverse metabolic markers in longer term OC users over the age of 35, in comparison to similar-age control groups, does not support the routine use of lipid and diabetes screening before initiating or during the use of low-dose OCs. PMID- 8233267 TI - Oral contraceptives and endometrial cancer. AB - OBJECTIVE: To evaluate the effect of ever-use of oral contraceptives (OCs) on the risk of endometrial cancer in women aged 50-64 years. METHODS: A case-control study was carried out using 142 incident cases of endometrial cancer and 1042 matched controls, aged 50-64. All subjects were members of Group Health Cooperative of Puget Sound in Seattle, Washington. Relative risk (RR) estimates were calculated for ever-users of OCs compared to never-users. RESULTS: Twenty six cases (18%) and 270 controls (26%) had taken OCs at some time, yielding an RR of 0.48 (95% confidence interval [CI] 0.26-0.89) for ever-users compared to never users. This estimate was adjusted for multiple risk factors. There was little effect of duration of OC use, age at first use, or number of years since last use in these data. CONCLUSION: Women aged 50-64 years at Group Health Cooperative who had taken OCs at some time appear to be protected from developing endometrial cancer later in life, compared to women who had never taken OCs. PMID- 8233268 TI - Ultrasonic thickness of the endometrium correlated to body weight in asymptomatic postmenopausal women. AB - OBJECTIVE: To estimate the prevalence and significance of an endometrial thickness of 5 mm or greater measured by ultrasound in postmenopausal women without vaginal bleeding. METHODS: Three hundred asymptomatic postmenopausal women scheduled for cervical cancer screening were also examined by transvaginal ultrasound. When the endometrium was 5 mm thick or greater, ultrasound was repeated every third month. Curettage was performed at the end of the study or if endometrial growth or vaginal bleeding occurred. RESULTS: The mean (+/- standard deviation) endometrial thickness was 2.3 +/- 1.8 mm (range 0-10). In women with endometrium measuring less than 5 mm, the endometrial thickness correlated to body weight. The endometrium measured 5 mm or more in 22 women, who were followed with ultrasound for 1-25 months. Body weight and body mass index (BMI) were higher in women with a thick endometrium. Serum levels of estrone, estradiol, FSH, and LH differed from those in women with normal ultrasound findings. Curettage revealed no case of malignancy. Fourteen cases of benign ovarian lesions were also diagnosed, and only one case of pelvic fluid was found (in a patient with liver cirrhosis). CONCLUSION: The prevalence of a thick endometrium was high in asymptomatic women, and endometrial thickness correlated with BMI. PMID- 8233269 TI - Local immune response in persistent cervical dysplasia. AB - OBJECTIVE: To investigate the differences in local immune response between persistent and regressive cervical dysplasia. METHODS: We conducted a quantitative study of Langerhans cells, pan-T cells, and helper-inducer T cells in the subepithelial connective tissue using immunohistochemical techniques with S-100 protein antibody, UCHL1, and OPD4, respectively, in 52 paraffin sections. The subjects were patients with persistent cervical dysplasia and a comparable control group of 46 patients with regressive dysplasia. RESULTS: In the subepithelial stroma, the mean (+/- standard deviation) numbers of S-100- and OPD4-positive cells in the persistent group were 8.6 +/- 8.1 and 84.6 +/- 66.3, respectively, compared with 15.1 +/- 9.4 and 147.0 +/- 67.7, respectively, in the regression group. These data demonstrate a significant reduction of Langerhans cells and helper-inducer T cells (P < .0002 and P < .0001, respectively) with persistent dysplasia. CONCLUSION: The decreased numbers of S-100-positive Langerhans cells and helper-inducer T cells in persistent dysplasia compared to those of regressive dysplasia strongly support a decreased local immune response in persistent cervical dysplasia. PMID- 8233270 TI - Macrophage colony-stimulating factor as a tumor marker for epithelial ovarian cancer. AB - OBJECTIVE: To determine the serum level of macrophage colony-stimulating factor in ovarian cancer patients in order to evaluate its role as a marker for ovarian cancer. METHODS: Serum macrophage colony-stimulating factor levels were assayed in 69 patients with epithelial ovarian cancer, 55 with benign ovarian tumors, and 634 healthy individuals, including 398 women, using an enzyme-linked immunosorbent assay. RESULTS: The average serum macrophage colony-stimulating factor level was 754.4 +/- 153.9 U/mL in healthy females; 1056 U/mL (mean plus 1.96 standard deviations) was considered to be the upper limit of normal. Serum macrophage colony-stimulating factor levels were significantly elevated in patients with ovarian cancer (average 1460.5 +/- 1006.2 U/mL; P < .001) and exceeded 1056 U/mL in 42 of the 69 patients with ovarian cancer (61%). No differences in levels were observed among the histologic types. No definite relationship was found between serum levels of macrophage colony-stimulating factor and those of CA 125. We found that 96% of the patients with ovarian cancer had high serum levels of macrophage colony-stimulating factor and/or CA 125 values. There was no significant difference in the levels of macrophage colony stimulating factor between patients with benign ovarian tumors and healthy controls. Only 7.3% of the group with benign tumors had levels exceeding 1056 U/mL. CONCLUSION: Macrophage colony-stimulating factor is a marker for ovarian cancer. Determination of serum levels can be useful in detecting ovarian cancer, particularly in combination with CA 125. PMID- 8233271 TI - Pain relief following day-case diagnostic hysteroscopy-laparoscopy for infertility: a double-blind randomized trial with preoperative naproxen versus placebo. AB - OBJECTIVE: To study the effects of preoperative naproxen on postoperative and post-discharge outcome and consumption of analgesics in patients undergoing diagnostic hysteroscopy and laparoscopy for infertility. METHODS: A double-blind, placebo-controlled study was conducted in 60 healthy women scheduled for day-case hysteroscopy and laparoscopy. Subjects were randomized to receive either 500-mg naproxen suppositories (n = 30) or placebo (n = 30) preoperatively. Following discharge, each patient was given two naproxen suppositories for treatment of pain at home. Pain was scored on the Visual Analogue Scale, and the postoperative and post-discharge use of analgesics was determined, recorded, and analyzed. RESULTS: Patients premedicated with naproxen had significantly less postoperative pain, showed more rapid ambulation, could be discharged earlier, and had less post-discharge pain. On the day after surgery, only six of 28 naproxen-treated patients needed analgesics, compared to 18 of 30 placebo patients. CONCLUSION: Day-case diagnostic hysteroscopy and laparoscopy for evaluation of infertility is potentially painful and stressful. Preoperative naproxen contributed to postoperative pain prevention, reduced hospital stay and consumption of analgesics, and shortened the period of post-discharge abdominal discomfort. PMID- 8233272 TI - Pregnancy planning and pre-conception counseling. The PRAMS Working Group. AB - OBJECTIVE: To estimate the percentage of women with one or more of four potentially modifiable risks who could have availed themselves of pre-conception counseling. METHODS: We defined pre-conception counseling to be consultation that occurs shortly before a couple attempts conception. Thus, we assumed that to obtain pre-conception counseling, a woman must plan her pregnancy. We used data from a population-based survey of 12,452 new mothers in four states who delivered babies during 1988-1990. Mothers were contacted 3-6 months after delivery and asked about pre-conception behaviors and the planning status of their pregnancies. We estimated the percentage of women who planned their pregnancies and had an indication for pre-conception counseling related to smoking, drinking, being underweight, or delaying initiation of prenatal care. RESULTS: State specific response rates ranged from 68-84%. Sixty percent of mothers reported that their pregnancies were planned. In general, mothers with unintended pregnancies were more likely to have an indication for pre-conception counseling than mothers with planned pregnancies. Thirty-eight percent of all mothers planned their pregnancies and had one or more indications for pre-conception counseling. An additional 30% had one or more indications for counseling but did not have a planned pregnancy. CONCLUSIONS: Despite the limited range of indications for counseling that we considered, a substantial percentage of women potentially could have used counseling. A similar percentage of women could have benefited from family planning services. PMID- 8233273 TI - The association between increased mean arterial pressure and abnormal uterine artery resistance to blood flow during pregnancy. AB - OBJECTIVE: To examine the relation between increased mean arterial pressure (MAP) and the uterine artery resistance index (RI) to blood flow in the third trimester of pregnancy. METHODS: One hundred consecutive patients in the third trimester whose pregnancies were complicated by hypertensive disorders, with or without proteinuria, had uterine artery resistance to blood flow measured using continuous-wave Doppler. RESULTS: The RI was in the normal range in 20 patients. Unilaterally abnormal RI was found in 32 and bilaterally abnormal RI in 46 (two subjects were excluded from this categorization because measurements were obtained from only one side of the uterus). A MAP below 100 mmHg was associated with a normal RI. All six subjects with MAP of 140 mmHg or greater had bilaterally abnormal RI. Mean arterial pressure values of 100-139 mmHg included 90% of the women with normal RI, 100% of those with unilaterally abnormal RI, and 87% of those with bilaterally abnormal RI. Proteinuric and nonproteinuric groups differed with respect to both MAP (P = .04) and RI (P = .01). Pregnancy outcome was less favorable in the unilaterally abnormal RI subgroup than in the normal RI subgroup. The bilaterally abnormal subgroup of patients had the most adverse pregnancy outcomes. CONCLUSIONS: Pregnant patients in the third trimester who have increased MAP but a normal uterine artery RI can expect a good pregnancy outcome. Increased MAP associated with abnormal uterine artery RI may result in an unfavorable pregnancy outcome, particularly in those with bilaterally increased RI to blood flow. Doppler measurement of uterine artery impedance is recommended in pregnancies complicated by hypertensive disorders. PMID- 8233274 TI - Pregnancy outcome in relation to uterine artery flow velocity waveforms and clinical characteristics in women with antiphospholipid syndrome. AB - OBJECTIVE: To determine whether uterine artery velocimetry is a useful tool for identifying pregnancies with antiphospholipid syndrome at risk for adverse outcome. METHODS: Twenty-four women with antiphospholipid syndrome, who had experienced 56 fetal losses in 63 previous pregnancies (88.9%), were treated with prednisone (40 mg/day) and aspirin (100 mg/day) during 28 pregnancies. Color Doppler ultrasound was performed at 18-24 weeks' gestation to investigate the resistance index of the uterine arteries. RESULTS: Treated women delivered 23 live infants in the 28 pregnancies (82.1%). Three infants weighed less than the tenth percentile (13%). Five pregnancies were complicated by preeclampsia and ten by nonproteinuric gestational hypertension. Positive results for all three assays for antiphospholipid antibodies (anticardiolipin antibodies, lupus anti coagulant, VDRL) at conception identified pregnancies destined to have poor fetal outcome and a significantly lower birth weight compared to pregnancies not having all three assays positive. An abnormal resistance index of the uterine arteries predicted pregnancies with poor fetal outcome in terms of week of delivery, birth weight, and birth percentile, as well as four of five cases of preeclampsia. CONCLUSIONS: Three assays positive for antiphospholipid antibodies at conception and an abnormal resistance index of the uterine arteries at 18-24 weeks' gestation predicted pregnancies at major risk for obstetric complications. Future studies should determine whether treatment can be modulated based on the Doppler findings. PMID- 8233275 TI - Uterine artery Doppler velocity waveforms in twin pregnancies. AB - OBJECTIVE: To compare uterine artery resistance index values in twin and singleton pregnancies, to examine eventual modifications of these values in twin pregnancies complicated by gestational hypertension and preeclampsia, and to determine whether resistance index values in twin pregnancies could predict the development of gestational hypertension and preeclampsia. METHODS: In a cross sectional study, reference limits for gestation were constructed for the uterine artery resistance index (higher, lower, and mean values) in 96 uncomplicated twin pregnancies and compared to the reference limits constructed from 315 normal singleton pregnancies. Uterine artery resistance indexes obtained in 53 twin pregnancies complicated by either gestational hypertension or preeclampsia were compared with the newly established nomograms. The clinical efficacy of the uterine artery resistance index to predict hypertensive complications was evaluated prospectively in 64 twin pregnancies studied at 20-24 weeks' gestation. RESULTS: In both singleton and twin pregnancies, uterine artery resistance indexes decreased linearly with advancing gestation. However, twin pregnancies showed significantly different slopes and constant values, resulting in lower resistance indexes at all gestational ages examined. No significant differences were found when comparing resistance indexes in all patients with gestational hypertension or preeclampsia to the reference limits. Statistically significant differences were obtained for the higher (P < or = .05) and mean (P < or = .01) resistance indexes when the comparison was restricted to preeclamptic patients. In the twin pregnancies studied at 20-24 weeks' gestation, the diagnostic efficacy of the uterine artery resistance index for predicting the development of gestational hypertension and/or preeclampsia was disappointingly low (k < 0.10). CONCLUSIONS: Resistance index values in the uterine artery are lower in twin pregnancies than in singleton pregnancies. Gestational hypertension and preeclampsia may occur in twin pregnancies despite normal uterine artery velocity waveforms, suggesting a limited role of this measurement in the management and prediction of hypertensive complications in twin pregnancies. PMID- 8233276 TI - Epidural anesthesia during labor and stress incontinence after delivery. AB - OBJECTIVE: To test the hypothesis that epidural anesthesia during labor prevents the development of stress incontinence after vaginal birth. METHODS: We interviewed 208 primiparas about stress incontinence 3 months postpartum. The women who developed stress incontinence after delivery were interviewed again 1 year postpartum. RESULTS: Twelve of 45 women (27%) who had epidural anesthesia developed stress incontinence de novo after vaginal delivery, versus 21 of 163 (13%) who did not receive epidural anesthesia, a marginally significant difference (P = .05). Those who had epidural anesthesia also had a significantly longer first stage of labor. One year after delivery, three of 42 (7%) in the epidural anesthesia group had stress incontinence, compared with five of 163 (3%) in the control group. CONCLUSION: Our results do not support the hypothesis that epidural anesthesia protects against the development of stress incontinence after vaginal delivery. PMID- 8233277 TI - Fetal platelet counts correlate with the severity of the anemia in red-cell alloimmunization. AB - OBJECTIVE: To determine whether fetal anemia secondary to maternal red-cell alloimmunization is associated with thrombocytopenia. METHODS: The records of 78 patients undergoing intrauterine transfusion for red-cell alloimmunization were reviewed. Pre-transfusion fetal platelet counts were compared between hydropic and nonhydropic fetuses. A regression analysis was performed between the fetal platelet counts and the fetal bilirubin levels, hematocrits, and reticulocyte counts taken at the initial transfusion. The hematocrits, reticulocyte counts, and bilirubin levels were adjusted for gestational age by calculating the number of standard deviations (SDs) from the mean for that age or the multiples of the mean (MOM). Student t test, Pearson coefficient, and contingency table randomization test were used to analyze the data. P < .05 was considered significant. RESULTS: Thirty-seven fetuses were hydropic and 41 were nonhydropic. Hydropic fetuses had a significantly lower platelet count than nonhydropic fetuses (197.5 +/- 86.4 versus 252.6 +/- 73.7 x 10(3)/microL; P < .01). Platelet counts correlated negatively with the reticulocyte count MOM (r = -0.652; P < .01) and the hematocrit SDs below the mean (r = -0.659; P < .01), but did not correlate with the bilirubin MOM (r = -0.183; P = .2). CONCLUSION: Hydropic and severely anemic fetuses are at increased risk for thrombocytopenia. We suggest that increased erythropoiesis diverts the hematopoietic stem cell away from thrombopoiesis. PMID- 8233278 TI - Fetal endothelin levels and placental vascular endothelin receptors in intrauterine growth retardation. AB - OBJECTIVE: To examine endothelin 1 (ET-1) in normal pregnancies and those complicated by severe intrauterine growth retardation (IUGR) with normal or increased fetoplacental vascular resistance as assessed by umbilical artery Doppler flow velocimetry. METHODS: Endothelin 1 concentrations were measured by radioimmunoassay in fetal umbilical arterial and venous and maternal venous plasma, and vascular ET-1 receptors were quantified by radioligand binding methods in placental primary and secondary stem villous vessels. RESULTS: Fetal and maternal ET-1 concentrations were similar in normal pregnancy (10-15 pmol/L) and were not influenced by mode of delivery (vaginal versus cesarean). Fetal levels were identical to control values in the growth-retarded fetuses with normal umbilical artery flow, but were markedly elevated (above 50 pmol/L) in two cases with increased fetoplacental vascular resistance. The corresponding maternal levels also appeared elevated, but to a lesser extent. Endothelin receptors were identified in placental vascular tissue. No differences in receptor-binding characteristics were observed between the groups. CONCLUSION: The increased fetoplacental vascular resistance present in a proportion of pregnancies complicated by severe IUGR may be associated with elevated fetal plasma ET-1 concentrations, but with no apparent change in vascular sensitivity to the peptide. PMID- 8233279 TI - Does Doppler-detected fetal movement decrease the incidence of nonreactive nonstress tests? AB - OBJECTIVE: To determine whether a decreased incidence of nonreactive nonstress tests (NSTs) in antepartum testing was attributable to the addition of fetal movement detection to the standard NST. METHODS: Monitors with standard fetal heart rate recording capabilities were used, as were new monitors producing a Doppler-detected recording of fetal movement (NST-fetal movement). Cross sectional retrospective analysis of NST results was carried out by chi 2. RESULTS: Comparison of the 10-month period before fetal movement detection to the 10 months including NST-fetal movement monitoring showed a significant decrease in nonreactive NSTs from 5.7% to 3.3% (chi 2 = 61.7, 95% confidence interval [CI] 0.97-0.98). This reduction in nonreactive tests disappeared (3.3% to 5.1%) when the NST-fetal movement-capable monitors were no longer available (chi 2 = 24.2, 95% CI 1.01-1.03). CONCLUSIONS: Nonreactive NSTs decreased by 58% with the introduction of fetal movement monitoring in our antepartum testing center and increased when the NST-fetal movement-capable monitors were removed. A reduced incidence of nonreactive NSTs associated with NST-Doppler-detected fetal movements should effect a savings in both time and resources. PMID- 8233280 TI - Antitumorous and immunomodulatory effects of the Viscum album L. preparation Isorel. AB - There are numerous data on the immunostimulative and antitumorous activity of various Viscum album tissue extracts. Isorel (Novipharm, Austria) is one of these compounds. We found that in mice an increased number of plaque-forming cells to sheep red blood cells (SRBC) followed the injection of Isorel together with SRBC. Further, survival time of a foreign skin graft was shortened if Isorel was applied at the correct time. Finally, suppressed immune reactivity in tumorous mice recovered following Isorel injection. Isorel was further shown to be cytotoxic to tumor cells in vitro. Its application to tumor-bearing mice could prolong their life but without any therapeutic effect. However, a combination of local irradiation and Isorel was very effective: following 43 Gy of local irradiation to a transplanted methylcholanthrene-induced fibrosarcoma (volume about 240 mm3) growing in syngeneic CBA/HZgr mice, the tumor disappeared in about 25% of the animals; the addition of Isorel increased the incidence of cured animals to over 65%. The combined action of Isorel, influencing tumor viability on the one hand and the host's immune reactivity on the other, seems to be favorable for its antitumor action in vivo. PMID- 8233281 TI - A study of the mechanisms involved in the immunostimulatory action of the pineal hormone in cancer patients. AB - The mechanisms responsible for the immunostimulatory role of the pineal hormone melatonin (MLT) are still obscure. To investigate the influence of MLT on interleukin-2 (IL-2)-induced immune effects in cancer, we compared the results obtained in 14 cancer patients treated with IL-2 (6 x 10(6) IU/day s.c. for 5 days/week for 4 weeks) plus MLT (10 mg/day orally) with those seen in 14 patients treated with IL-2 alone and with those obtained from 14 other patients treated with MLT only. All patients were affected by metastatic solid neoplasms. The increase in the mean number of lymphocytes, T lymphocytes, natural killer cells, CD25-positive cells and eosinophils was significantly higher in patients treated with IL-2 plus MLT than in those receiving IL-2 alone. On the contrary, the increase in mean serum levels of the macrophage marker neopterin was significantly higher in patients treated with IL-2 alone than in those treated with IL-2 plus MLT. Finally, MLT alone has no significant effect on immune cell mean number and on neopterin secretion. These results would suggest that the immunostimulatory action of MLT requires the concomitant presence of IL-2 and that two of the main target cells for MLT activity in humans are represented by T helper lymphocytes of type 2, which are involved in IL-2-induced eosinophilia by the release of IL-5, and macrophages, which may inhibit IL-2-dependent immune functions. PMID- 8233282 TI - Utility of serum activity of angiotensin-converting enzyme as a tumor marker. AB - The objective of this study was to evaluate the diagnostic utility of the measurement of the serum activity of angiotensin-converting enzyme (SACE) as a cancer marker. This case-control study included 135 patients with cancer of different sites, confirmed histologically, and 145 controls (107 normal individuals plus 38 chronically ill patients with nontumoral diseases). Determination of SACE activity was done by a spectrophotometric method using as substrate the synthetic tripeptide N-(3-[2-furyl]acryloyl)-L-phenylalanylglycine. There were no sex- or age-related variations in SACE activity. Mean SACE activity in 107 normal controls was 51.6 U/l (95% C.I., 50.1-53.1); in 145 nontumoral individuals, including 38 chronic nonmalignant diseases plus 107 normal controls, 51.5 (50.1-53.1) and in malignant tumors 35.7 (32.8-38.5). There was no statistically significant difference between chronic diseases and normal controls (p > 0.05); but there was one between cancer patients and nontumoral individuals, normals and chronic nontumoral diseases. The mean of SACE activity values by tumoral site are (U/l; 95% C.I.): breast, 41.3 (36.2-46.5); gastrointestinal 31.5 (24.3-38.8); head and neck, 32.3 (26.7-37.8), and lung 27.6 (21.6-33.6) (p < 0.001). The means by clinical stage are: complete remission, 58.0 (53.7-62.3), significantly higher than in normal controls (p < 0.001); local disease, 40.56 (34.5-46.5); locoregional disease, 35.09 (30.7-39.4); metastatic disease, 23.04 (19.5-26.5), and in relapse at diverse stages, 30.86 (25.1-36.5). In clinical active cases, there is a statistically significant decrease of SACE activity, especially in metastatic disease (p < 0.001). The calculated cutoff value, excluding complete remission cases, is 40.7 U/l, with sensitivity of 69.5% and specificity of 91.6%. We conclude that there is a decrease of SACE activity in cases of clinically active cancer and an increase in clinical complete remission. PMID- 8233283 TI - Clinical utility of TPS, TPA and CA 19-9 measurement in pancreatic cancer. AB - The aims of this study were to (1) evaluate the diagnostic utility of a new tumor marker, TPS, with respect to TPA and CA 19-9 in patients with pancreatic cancer; (2) ascertain the reliability of the markers in predicting survival, and (3) evaluate the effect of liver dysfunction on the results. CA 19-9, TPA and TPS were measured in the serum of 19 control subjects, 42 patients with pancreatic cancer, 29 with chronic pancreatitis, and 52 with extrapancreatic diseases. CA 19 9 was confirmed to be the best serological indicator of pancreatic cancer, while TPA and TPS lacked both sensitivity and specificity. Pancreatic cancer patients with liver metastases had higher mean CA 19-9 and TPA, but not TPS, values than pancreatic cancer patients without metastases. A shorter survival time was associated with the presence of liver metastases and with higher serum tumor marker levels. CA 19-9, TPA and TPS were found to be correlated with liver function test results (ALT, ALP and bilirubin). IN CONCLUSION: (1) TPS adds no significant information to that obtained using CA 19-9 in the diagnosis of pancreatic cancer; (2) CA 19-9 and TPA, but not TPS, are influenced by the presence of liver metastases; (3) the main factor to influence survival is advanced disease, which is in turn associated with higher tumor marker levels, and (4) liver dysfunction can influence not only CA 19-9 and TPA, as already described, but also TPS. PMID- 8233284 TI - Symptomatic cardiotoxicity with high-dose 5-fluorouracil infusion: a prospective study. AB - Cardiotoxicity with 5-fluorouracil (5-FU) therapy has been reported to range from asymptomatic EKG abnormalities to fatal myocardial infarction. We report a prospective study in 100 consecutive patients receiving 5-FU infusion in combination with other chemotherapeutic agents or alone for the treatment of different malignancies with the aim of identifying patients who develop symptomatic cardiotoxicity. Patients with a history of cardiac illness, abnormal EKG or cardiac enzyme levels were excluded. Patients were observed during the total period of infusion, daily EKG was performed on asymptomatic patients, those who developed cardiotoxicity were monitored till symptom-free for 24 h. Eight patients developed symptoms suggestive of cardiotoxicity. Pain was the commonest symptom (5/8), followed by palpitation and sweating. Three patients developed EKG abnormalities and 1 went into cardiogenic shock. Time to toxicity ranged from 18 to 30 h (mean 24 +/- 3.7 h) and serial cardiac enzyme levels remained normal in all patients. The symptoms reversed immediately on cessation of the treatment in most of the patients (7/8). Time to recovery ranged from 5 to 60 min (mean 19.28 +/- 19.6 min). There was no recorded death due to toxicity. We conclude that 5-Fu infusion is associated with a significant risk of symptomatic cardiotoxicity. Concomitant chemotherapeutic agents, received by all the affected patients, may have a contributory effect too. Cardiotoxicity seems to be completely reversible, particularly in patients without underlying cardiac disease. The patients should be informed about the symptoms and the condition recognised and managed immediately. PMID- 8233285 TI - Arm morbidity following treatment of breast cancer with total axillary dissection: a multivariated approach. AB - In order to clarify the factors that mainly influence arm morbidity following treatment of breast cancer with the full axillary dissection protocol, we evaluated, in a model of multiple regression analysis, parameters such as the type of breast surgery, adjuvant radiotherapy, time of irradiation, age, number of dissected nodes and axillary nodal status. A total of 104 women were studied. Late arm edema was observed in 17% of the patients and was more frequent when (1) irradiation was given immediately after the operation than if it was given 6 months later (p = 0.009) and (2) the number of removed nodes exceeded 40 (p = 0.037). Upper limb pain was reported by 16% of the patients and was reported more frequently from patients over 60 years of age (p = 0.036), as well as from patients who underwent modified radical mastectomy (p = 0.044) and those in whom 30-40 nodes were dissected (p = 0.025). Shoulder joint mobility was impaired in 17% of the patients, and it was not affected by any of the examined factors. It seems that conservative breast surgery or adjuvant breast radiotherapy 6 months after the operation might reduce independently the likelihood of arm morbidity by 25%. PMID- 8233286 TI - Prophylaxis of superficial bladder cancer with a modified intravesical epirubicin treatment schedule. AB - A controlled prospective trial on 99 eligible patients evaluated the efficacy of intravesical epirubicin administration as prophylaxis against tumor recurrences after complete endoscopic resection of superficial bladder cancer. The treatment schedule consisting of an initial 6- or 8-week course of instillations, followed by single maintenance doses (to the responders) at follow-up examinations, was modified in those of the initial responders who were at high risk for recurrence and who received an additional separate 4-week course of therapy. Sixty percent of the epirubicin-treated patients and 41% of the controls (who underwent resection alone) remained free of recurrences for a mean follow-up of 32.1 months, but the difference was not significant. However, in terms of relative risk for recurrences (recurrence rate per 100 patient-months and disease-free interval), comparisons between the two groups of patients revealed a significant benefit of epirubicin, overall as well as for those with a history of previous tumor recurrences, multifocal disease, and stage Ta and grade 2 neoplasms. Drug induced toxicity was acceptable. PMID- 8233287 TI - Propofol improves patient comfort during cisplatin chemotherapy. A pilot study. AB - 20 consecutive patients with nausea and vomiting secondary to cisplatin chemotherapy uncontrolled by serotonin-antagonist and corticosteroid prophylaxis during their first cycle received adjuvant propofol. This new anesthetic agent was added at subhypnotic doses, i.e. 1 mg/kg/h, as a continuous intravenous infusion during the two subsequent chemotherapy cycles. In 85 and 90% of patients, nausea and vomiting were prevented in the first 24 h following the first and second propofol-supplemented chemotherapy cycles respectively. 24-72 h postchemotherapy, this side effect remained suppressed in 75 and 70% of patients, respectively. Patients' comfort and appetite were improved. All 20 subjects preferred the propofol-containing regimen. PMID- 8233288 TI - Analysis of time to response to chemotherapy in 316 metastatic breast cancer patients. AB - Chemotherapy is a major tool for metastatic breast cancer treatment. In this study, a series of 316 patients have been analyzed to evaluate the time needed to reach tumor response by means of combination chemotherapy. Twenty-five percent of patients responded within 3 months and virtually all responses occurred within 7.5 months. The time curves of response (any) and best response are superimposable. A subset analysis has shown that the following pretreatment characteristics predict a significantly longer time to response: prior exposure to adjuvant chemotherapy, nodal positivity at diagnosis, no previous endocrine treatment and osseous metastases. PMID- 8233289 TI - Effect of various doses of intravenous polyclonal IgG on in vivo levels of 12 pneumococcal antibodies in patients with chronic lymphocytic leukaemia and multiple myeloma. AB - Polyclonal intravenous IgG (IVIG) was administered as an infusion 6 times every 3 weeks (week 0, 3, 6, 9, 12, 15) in doses of 0.1, 0.4 and 0.8 g/kg BW to determine the dose causing an increase in 12 pneumococcal antibody types above the protective level of 200 ng/ml of antibody N. The dose of 0.4 g/kg BW was found to be optimal in patients with chronic lymphocytic leukaemia (CLL). From the first infusion onwards at least 80% of CLL patients had increases in all 12 antibodies. Five weeks after the last infusion the antibody levels were still elevated in 80% of patients with CLL. The dose of 0.8 g/kg raised all 12 antibodies in 53-73% of CLL patients when assessments were made after each infusion. In multiple myeloma (MM) patients, 73-82% and 73-91% of patients had increased antibody levels, respectively, before and after the 4th-6th infusions at the 0.8 g/kg dose level. However, in only 45-50% of patients did the antibodies remain increased 2 weeks after the treatment at this dose. The dose of 0.4 g/kg caused antibody increases in only 30-50% of patients when measured before the 4th-6th infusion. Serum IgG increased significantly only in the CLL patients, whereas in the MM patients it was high from the beginning owing to the disease. Therefore, the pneumococcal antibody levels were a better marker for the purpose of dose finding. The dosage recommendation in CLL is 0.4 g/kg every 3 weeks until week 12, when steady state is reached. The maintenance dose is 0.4 g/kg every 5 weeks. In MM patients, who have a faster elimination rate of antibodies, the recommended loading dose is 0.8 g/kg, followed by 0.4 g/kg every week as a continuous treatment. Treatment with IVIG in CLL and MM was generally well tolerated. Only 25% of patients experienced minor side-effects, the most frequent being febrile reactions, shivering and headache. PMID- 8233290 TI - The effect of minocin on the pericardium. AB - The purpose of this study was to evaluate in a dog model whether intrapericardial instillation of tetracycline is superior to the effect of drainage alone in causing pericardial adhesions and cavity obliteration. Twelve mongrel dogs were randomly divided into two experimental groups. All dogs received a pericardial drain through a sterile thoracotomy. Group A dogs (n = 6) received minocycline hydrochloride, 20 mg/kg, group B received normal saline. All dogs were sacrificed 1 month later. Echocardiograms performed 1 week postsurgery were normal in all dogs. Macroscopic evaluation disclosed that all group A dogs had over 25% cavity obliteration whereas group B dogs had no adhesions or had obliteration of less than 25% of the cavity area. Microscopic evaluation showed that group A dogs had severe pericardial fibrosis and thickening with slight focal lymphoplasmocytic infiltration. Myocardial damage was not seen. Our results support the clinical use of injecting tetracycline into the pericardium (in addition to drainage) to cause cavity obliteration in patients with persistent pericardial effusion. PMID- 8233291 TI - Treatment with tamoxifen and progestins for metastatic breast cancer in postmenopausal women: a quantitative review of published randomized clinical trials. AB - The efficacy of tamoxifen (TAM) was compared to that of progestins (medroxyprogesterone acetate, MPA, and megestrol acetate, MA) in the treatment of metastatic breast cancer in postmenopausal women by a quantitative analysis of the results of published randomized clinical trials. Seven studies involving a total of 801 subjects compared TAM with MPA. Overall, the frequency of complete and partial response was 9 and 18%, respectively, in the women treated with TAM, versus 9 and 28% in those given MPA. Considering complete and partial responses together, the frequency of response was 29% in the TAM group and 39% in the MPA group, the corresponding pooled odds ratio (OR) of response being 1.5 (95% confidence interval, CI, 1.1-2.0). The median duration of response was greater in the TAM-treated patients; however, the difference was small (14 vs. 11 months). The probability of response to MPA treatment was about 3-fold higher compared with the response to TAM treatment in the subgroup with bone metastases (OR 3.4), and 2-fold higher in the subgroup with visceral metastases (OR 2.2), but the difference in the OR estimates was not statistically significant. The response to the two drugs was similar in the subgroup with metastases in soft tissues. Four studies compared TAM with MA, taking in 463 subjects. The overall frequency of complete and partial response was 35% in the patients who received TAM compared with 29% in those treated with MA. The corresponding pooled OR was 0.8 (95% CI 0.5-1.1). Analysis of the results according to site of metastases revealed no significant difference in the frequency of complete or partial response in the two treatment groups. PMID- 8233292 TI - A phase II study of levofolinic acid and 5-fluorouracil plus cisplatin in patients with advanced head and neck squamous cell carcinoma. AB - Forty patients with advanced squamous cell carcinoma of the head and neck (SCHNC) were treated with a combination of levofolinic acid 100 mg/m2+5-fluorouracil 375 mg/m2 in a 4-hour infusion plus cisplatin 20 mg/m2 in a 2-hour infusion for 5 consecutive days, repeated every 21-28 days. In the group of 20 previously untreated patients, a 90% overall response rate (ORR) with a 30% complete response rate (CRR) was obtained. In the group of 20 pretreated patients with recurrent and/or metastatic SCHNC, a 55% ORR with 15% CRR was achieved. This treatment was given on an outpatient basis and was generally very well tolerated with only 2 patients requiring hospitalization. Grade 1-2 gastrointestinal and hematological side effects were the most frequent toxicities. One patient had grade 4 liver toxicity, 1 had grade 4 anemia, and 1 grade 3 neurotoxicity. This treatment seems very active in both previously untreated and pretreated patients. However, in the latter group the mean duration of complete response (12.2+ months) and of partial response (7.4+ months) are, in our opinion, still unsatisfactory. PMID- 8233293 TI - The clinical significance of the histologic subclassification of colorectal carcinoma. AB - According to the histologic features at the deepest level of tumor invasion (the tumor apex), we classified colorectal carcinomas as follows: well differentiated (W), moderately differentiated (M), and mucinous (Muc). By assessing its glandular configuration and cellular arrangement, the M type was further subdivided in to two different groups: moderately well differentiated (Mw) and moderately poorly differentiated (Mp) carcinomas. In our sample there were 56 W, 53 Mw, 20 Mp and 13 Muc tumors. The malignant potential of each tumor was evaluated by examining for the presence or absence of lymphatic invasion, venous invasion, lymph node metastases and liver metastases. In comparison with the other tumors, the Mp tumors proved to harbor the highest malignant potential, exhibiting a higher incidence of lymphatic invasion (95%, 19/20 of the cases), venous invasion (75%, 15/20 of the cases), lymph node metastases (80%, 16/20 of the cases), and liver metastases (40%, 8/20 of the cases). Colorectal carcinomas are composed of multiple cell populations with different biological and malignant properties, and through the histologic subclassification we could identify that Mp tumors have the highest potential to invade normal tissue and metastasize. PMID- 8233294 TI - Use of the succinate dehydrogenase inhibition test in assessing the heat sensitivity of tumor cells. AB - The in vitro succinate dehydrogenase (SD) inhibition (SDI) test was used to determine the heat sensitivity of tumor cell lines and human tumor cells. The tumor cells were exposed to heat in vitro and the decrease in SD activity was assayed using a colorimetric assay, the SDI test. With respect to survival curves of HeLa cells, the SDI test correlated well with the clonogenic assay and the dye exclusion assay. Decrease in the SD activity of HeLa cells after the heat treatment (41-44 degrees C, 10 min to 5 h) depended on both the temperature and the duration of heat exposure. S- and G2/M-phase-rich HeLa cells were more sensitive to heat than were the G1-phase-rich cells. The SDI test exhibited a wide variation in the heat sensitivity among four cell lines (HeLa, B-16, V-79, and a human GT-1 squamous cell carcinoma). Variation in heat sensitivity was also detected among individual tumor tissues obtained from clinical specimens of gastric, esophageal, and colorectal cancers. Gastric cancer tissues were more sensitive to heat than were esophageal and colorectal tissues. We recommended that if the SDI test is used to assess the heat sensitivity of clinical tumor tissues in vitro, appropriate therapy for individual patients can be designed. PMID- 8233295 TI - CVM versus ACE in the treatment of small cell lung cancer. AB - In an attempt to achieve adequate palliation in patients with small cell lung cancer (SCLC) while keeping toxicity to a minimum, we compared CVM (carboplatin/vinblastine/methotrexate) and standard therapy with ACE (doxorubicin/cyclophosphamide/etoposide). None of the 104 SCLC patients with limited or extensive disease who participated had received previous treatment. After stratification according to disease extent, patients were randomized to receive either CVM or ACE. The maximum number of chemotherapy courses was six. When response rates were compared, ACE was found to be somewhat superior to CVM in terms of objective response [CVM 67%, 95% confidence interval (CI) 54-79%; ACE 88%, 95% CI 80-97%; p = 0.06]; however, a significant difference was evident only among extensive-disease patients. Median response durations (CVM 6 months, 95% CI 5-8; ACE 5 months, 95% CI 3-6) and median survival times (CVM 8 months, 95% CI 7 10; ACE 7 months, 95% CI 4-9) were comparable. CVM met the goal of producing significantly less hematologic toxicity than occurred with ACE. Leukopenia affected 92% of ACE-treated patients and 48% of CVM-treated patients (p = 0.005), and was severe in 80% of the ACE group and 20% of the CVM group. Alopecia occurred much more frequently among those treated with ACE (91 vs. 24%; p < 0.001), as did infection (59 vs. 24%; p < 0.001). The selection of a specific chemotherapy regimen must be individualized. CVM may be appropriate for patients in whom intensive chemotherapy is contraindicated due to performance status, age, concomitant medical disease, or patient refusal. PMID- 8233296 TI - Combined radiochemotherapy with carboplatin in the treatment of advanced head and neck carcinomas. AB - From 1987 to 1991, 100 evaluable patients with advanced head and neck carcinomas (T2-4, N0-3) were treated with radiotherapy and simultaneous carboplatin. Tumors were located in the oral cavity in 33 patients, oropharynx in 8 patients, and hypopharynx in 7 patients. Four patients had a tumor of the epipharynx, 3 of the larynx, and 45 had involvement of two or more compartments. Radiotherapy was performed in a fractionation of 5 x 2 Gy/week up to a dose of 50 Gy. Carboplatin was administered in a dose of 60 or 70 mg/m2 from days 1-5 and 29-33. After a 2 week interval, tumor involution was evaluated and a decision was made on the patients' operability. In cases of inoperability, radiotherapy was continued up to a dose of 70-74 Gy. Thirty patients underwent surgery after 50 Gy. Eight patients showed a histologically complete remission (CR), 7 showed microscopic residual tumor, and 15 showed macroscopic tumor. Seventy patients were treated with radiotherapy and concomitant carboplatin only. Thirty-nine of them achieved a CR and 30 a partial remission (PR). The residual tumor was operable in 8 of the latter patients. Only a minor response was achieved in the remaining patient. At the end of the treatment 77 patients achieved a CR with this combined modality. From 1990 to 1992, 20 patients with locally advanced head and neck carcinomas underwent hyperfractionated accelerated radiotherapy (2 x 1.6 Gy/day, 5 days per week: total dose, 64-67.2 Gy) and simultaneous intravenous carboplatin (60 mg/m2, days 1-5 and 29-33) in a pilot study. Fifteen patients had T4 and 5 had T3 tumors. Six weeks after the end of treatment, 16 patients (80%) had CR, and PR was seen in the other 4 patients (20%). Overall and disease-free survival at 1 year was 82 and 81%, respectively. Although acute side effects were more pronounced compared with conventional irradiation, this treatment regimen is feasible and the initial CR rate of 80% is encouraging. Because of the results achieved with hyperfractionated accelerated radiotherapy, we initiated a multicenter randomized study in November 1991. Patients with advanced head and neck carcinomas are either randomized for conventional radiotherapy with carboplatin or hyerfractionated accelerated irradiation with carboplatin. Results will be forthcoming. PMID- 8233297 TI - Results of a Southwest Oncology Group phase III trial of carboplatin plus cyclophosphamide versus cisplatin plus cyclophosphamide in advanced ovarian cancer. AB - Cisplatin combined with cyclophosphamide has been considered a very active treatment for advanced ovarian cancer. Unfortunately, cisplatin is associated with dose-limiting neurotoxicity, as well as possible neuropathy, ototoxicity, and occasional renal dysfunction. Carboplatin, a cisplatin analogue, is active against advanced ovarian cancer, with a presumed lower incidence of emesis, ototoxicity, neuropathy, and renal dysfunction. The Southwest Oncology Group initiated a phase III randomized trial, in which 342 patients with stage III (suboptimal disease) and stage IV ovarian cancer were randomly assigned to treatment with six courses of intravenous cisplatin 100 mg/m2 plus cyclophosphamide 600 mg/m2 or carboplatin 300 mg/m2 plus cyclophosphamide 600 mg/m2. The median survival for the cisplatin arm was 17.4 months; for the carboplatin arm, median survival was 20.0 months. The null hypothesis of a 30% survival superiority with the cisplatin arm was rejected at the p = 0.02 level. Clinical response rates were 52% for the cisplatin arm and 61% for the carboplatin arm. There was less thrombocytopenia in the cisplatin arm (p < 0.001); however, there was less nausea and emesis (p < 0.001 for courses one to five), renal toxicity (p < 0.001), anemia (p < 0.001), hearing loss (p < 0.001), and neuromuscular toxicity (p < 0.001) in the carboplatin arm. Carboplatin/cyclophosphamide proved to have a significantly better therapeutic index than cisplatin/cyclophosphamide in these patients with advanced ovarian cancer. PMID- 8233298 TI - Neoadjuvant chemotherapy with carboplatin/5-fluorouracil in head and neck cancer. AB - In a prospective, randomized, multicenter study, neoadjuvant chemotherapy (CT) with carboplatin and 5-fluorouracil (5-FU) followed by locoregional treatment (LRT) was compared with locoregional treatment alone in the treatment of patients with head and neck cancer. This study, which includes 324 patients, was conducted from January 1988 to July 1991. The aim of this study was to evaluate the impact of the carboplatin/5-FU regimen both on the incidence of mutilating surgery and on the survival rate. Chemotherapy consisted of three cycles of carboplatin 400 mg/m2 day 1 and 5-FU 1 g/m2 days 1-5, repeated every 3 weeks. Patients with a complete tumor response then received radiotherapy alone, instead of the treatment planned initially. Three hundred patients were analyzed: 79 had tumors of the oral cavity, 106 oropharyngeal tumors, and 115 pharyngolaryngeal tumors. One hundred fifty patients underwent CT+LRT; 150 patients had LRT alone. Grade 3 and 4 toxicity rates were minimal in the CT+LRT group; toxicity was mainly hematologic (24% neutropenia, 19% thrombocytopenia). There were 3 toxic deaths (2%), 2 due to septicemia and 1 due to cardiac toxicity. One hundred forty-three patients were evaluable for efficacy. The tumor objective response rate was 63% and complete response rate was 31% (35% for oropharyngeal, 34% for pharyngolaryngeal tumors, 22.5% for oral cavity tumors), which led to a 29% decrease in the rate of mutilating surgery. Conservative treatment was performed in 57% of patients in the CT+LRT group vs. 24% in the LRT group (p = 0.001). There was no significant difference between survival curves in the CT+LRT and LRT groups. At 4 years, overall survival rates were 56 and 46%; disease-free survival rates were 33 and 30% in the CT+LRT and the LRT groups, respectively. The survival rates were not significantly different in the two groups. The locoregional recurrence rates were 35% in the CT+LRT arm and 25% in the LRT arm (p = 0.04), with median follow-up of 25 months. The rates of secondary localization and distant metastasis were not significantly different in the two groups. PMID- 8233299 TI - Carboplatin and urothelial tumors. AB - The prognosis of advanced-stage bladder cancer is poor. Chemotherapy, particularly regimens including platinum salts, appears to increase survival moderately but at the cost of severe, mainly renal toxicity. Platinum is a major factor in this toxicity, and new platinum salts (chiefly carboplatin) have therefore been developed. Carboplatin has no renal toxicity at usual doses, and its use does not require concomitant hyperhydration. Its gastrointestinal, otologic, and general tolerability is excellent. In contrast, most patients develop thrombocytopenia, which can be important, but which is always transitory. The platelet count reaches its nadir (grade 2 or 3) at around day 20, and the leukocyte nadir (grade 2 or 3) occurs about day 19. Anemia is rare. The literature on the use of carboplatin for the treatment of advanced-stage urothelial tumors is reviewed. Carboplatin is used at doses varying between 200 and 400 mg/m2, administered in 28-day courses. Dose adjustment is based on serum creatinine level, creatinine clearance, nadir blood cell levels, or previous treatment, reflecting the wide disparity between different studies. Used alone, carboplatin achieved objective responses (ORs) in 14% of patients (3% complete responses, CRs, and 11% partial responses, PRs) in a total group of 327 patients included in 13 trials. In polychemotherapy various combinations of carboplatin with other agents have been reported, most frequently carboplatin/methotrexate/vinblastine; the OR rate was 63% (CR rate 19% and PR rate 44%) among 88 patients in four studies. These results confirm the relative efficacy of carboplatin in the treatment of advanced-stage urothelial tumors, particularly when it is combined with other agents. Its efficacy is similar to that of cisplatin, but it is far less toxic. A prospective, comparative trial will be necessary to confirm these data. The pharmacokinetic behaviors of the two platinum salts are markedly different, as carboplatin does not undergo tubular metabolism. The efficacy of carboplatin could be optimized by adapting the dosage to the glomerular filtration rate, which is a more accurate method than extrapolation from the serum creatinine or creatinine clearance values. This has been shown in the case of nonseminomatous germ cell tumors. Calculation of the optimum carboplatin dose should now be applied to urothelial tumors. The general and renal tolerability of a platinum salt is an important element of choice when the efficacies are equivalent. These considerations fully warrant further clinical trials of carboplatin. PMID- 8233300 TI - Carboplatin in the treatment of non-small cell lung cancer: a review. AB - Cisplatin is among the most active agents available for the therapy of non-small cell lung cancer (NSCLC). Its congener, carboplatin, has been the focus of intense clinical trials over the last several years because of its improved therapeutic index. Results from randomized trials have shown that carboplatin is active in NSCLC, and that it can be effectively integrated into combination chemotherapy programs, with survival benefits comparable to cisplatin-containing regimens. Ongoing trials address critical issues, including: the optimization of platinum dose by either varying the carboplatin dose or combining cisplatin and carboplatin; evaluating the activity of carboplatin-containing programs with stem cell support; and combining low-dose carboplatin with radiation therapy in an attempt to integrate these effective modalities. Finally, the availability of new active agents such as TAXOL (paclitaxel) and the camptothecin analogs has led to clinical trials to determine the optimal interaction between these agents and carboplatin. PMID- 8233301 TI - High-dose carboplatin in the treatment of hematologic malignancies. AB - Carboplatin is a second-generation platinum complex developed to be less ototoxic and nephrotoxic than cisplatin. The major toxicity was found to be myelosuppression; thus, it was tried in acute leukemia. When given by daily bolus injection for 5 days, carboplatin exhibited some activity but was associated with additional nonhematologic toxicity as well. When administered by continuous infusion, responses were higher and toxicity less. The Eastern Cooperative Oncology Group (ECOG) conducted a phase II study of carboplatin 315 mg/m2 daily given by continuous infusion for 5 days to adults with refractory and relapsed acute leukemia. A second course was given if the bone marrow on day 14 revealed persistent leukemia. Those achieving a complete remission (CR) were given an additional course as consolidation. The median age was 49 years among acute myelogenous leukemia (AML) patients and 46 years in acute lymphoblastic leukemia (ALL) patients. Of 46 eligible patients enrolled in the study (36 AML and 10 ALL), 8 (17%) achieved a CR (6 AML and 2 ALL). Remissions were observed in 2 of 10 primary refractory patients (1 AML and 1 ALL). When treated in first relapse, 5 of 14 patients (36%) achieved a CR. In 38 instances marrow specimens were examined after treatment; 10 (26%) showed no change, 16 (42%) were hypoplastic, and 12 (32%) were hypoplastic with residual leukemic cells. Of the 18 deaths that occurred on study, 14 were due to infection, 2 due to infection and bleeding, 1 due to uncontrolled gastrointestinal bleeding and 1 due to graft-versus-host disease in a patient who had relapsed after bone marrow transplantation. Marrow suppression was usually prolonged. Nonhematologic toxicity was mild. Gastrointestinal toxicity consisted of easily controlled nausea and vomiting. Three patients had grade 3 diarrhea. Grade 3 or more renal toxicity was observed in 8 patients, all of whom had received nephrotoxic antibiotics for treatment of bacterial or fungal infections. One patient died of renal failure that developed near the end of a second induction course. Ototoxicity was observed in 11 patients (24%) and was grade 2 or less in all but 3. These results indicate that carboplatin is an active agent in leukemia. Further studies are under way in combination with other agents such as etoposide, mitoxantrone, 5-azacytidine, and daunorubicin in treatment of acute leukemia and in combination with ifosfamide and etoposide in refractory lymphomas. PMID- 8233302 TI - Escalating high-dose carboplatin and autologous bone marrow transplantation in solid tumors. AB - Twenty-seven patients with poor-prognosis malignancies were treated with a combination (CARBOPEC) of fixed-dose etoposide (1,750 mg/m2), cyclophosphamide (6,400 mg/m2), and escalating doses of carboplatin (from 800 to 1,600 mg/m2) followed by autologous bone marrow transplantation (ABMT). All patients had previously received platinum derivatives. The diagnoses were as follows: germ cell tumors (GCTs; n = 15); ovarian carcinomas (n = 8); rhabdomyosarcomas (n = 3), and Hodgkin's disease (n = 1). All 27 patients were fully evaluated for toxicity. The median duration of granulocytopenia (leukocytes < 0.5 x 10(9)/l) and thrombocytopenia (platelets < 20 x 10(9)/l) was 23 and 20 days, respectively. Hematologic growth factors were used in 3 cases. The main nonhematologic toxicity was gastrointestinal, with moderate to severe diarrhea in 18 patients. No significant renal toxicity was observed. The overall response rate to this high dose chemotherapy was 55%, with a complete response (CR) rate of 45% (9 patients). The median duration of CR was 9 months. Five of the 27 patients are alive with no evidence of disease (NED) at 5, 22, 27, 40, and 43 months after ABMT. Four of the 11 patients with refractory GCTs have NED at 5, 22, 27, and 40 months, together with 1 of the 3 responders (43 months). Our study shows the encouraging antitumor activity of this regimen. Similar chemotherapy schedules have also been used with high response rates in GCT, ovarian cancer, breast cancer, and soft tissue sarcoma in children. The CARBOPEC protocol seems to be a good candidate for therapy intensification in patients with various malignancies. A European trial for salvage therapy in GCT will be activated in the near future. Moreover, results should improve with the widespread use of hematopoietic growth factors and optimization of carboplatin administration that takes pharmacokinetic parameters into account. PMID- 8233303 TI - [Genetic disorders in the pollen cells of waxy barley in radioactive contamination after the Chernobyl accident]. AB - Genotoxic effects during microsporogenesis and gametogenesis were studied in barley of the waxy strain grown on 3 experimental plots situated in the region of the Chernobyl disaster (plot 1 in the town of Chernobyl, plots 2 and 3 in the town of Yanov, 3 km away from the damaged power plant). It was determined that an increased level of radionuclide pollution and chronic irradiation during ontogenesis results in higher incidence of meiotic disturbances and abnormalities during formation of the male gametophyte. Incidence of waxy reversions in pollen grains depends on the activity of radionuclide pollutants as well. We suggest that selection affecting both diplont and haplont forms will provide for the absence of significant aberrations in subsequent plant generations. PMID- 8233304 TI - [The sorting of heterotypic cells in mixed cultures]. AB - Bovine tracheal epithelium, FBT cells, transformed mouse kidney epithelium, MPTR cells, and normal mouse embryo fibroblasts were used to demonstrate cell sorting in mixed monolayer cultures grown on solid substrates. Sorting was caused by contact cell competition for the substrate territory. Central position after sorting was occupied by most adhesive cells capable of displacing other groups of cells from the substrate. PMID- 8233305 TI - [The sex dependence of the expression of the particular estrogen-binding protein in the hepatocytes in rat ontogeny]. AB - Formation of dependence on sex of the unusual estrogen-binding protein (UEBP) expression in rat liver during postnatal development was studied using radioligand and immunohistochemical techniques. UEBP was shown to appear in the male and female liver by day 45 to 50 of postnatal life. In females, UEBP remained at a low level until day 75 to 80, corresponding to faint UEBP-specific staining of a single layer of hepatocytes surrounding the central vein. Thereafter UEBP level decreased below the resolving capacity of the method. In males, UEBP content progressively increased during subsequent stages of ontogenesis. This is accompanied by formation of a stable descending centrolobular-periportal gradient of UEBP-positive hepatocytes. High UEBP expression in the male liver was found to result from the programming effect of androgens exerted between days 1-3 and 12 of postnatal development. The formation of hepatic lobuli is not a condition sufficient for switching on the androgen program of UEBP expression. The absence of testicular androgens in males after day 12 of life does not affect the androgen-programmed ontogenetic time course of UEBP expression at high levels. Experimental programming of UEBP expression in hepatocytes of immature females with the use of androgens results in lower UEBP levels than in males. These levels are comparable to those found in male liver at early stages of regeneration (96 h) after partial hepatectomy. It is suggested that androgens determine not only UEBP-producing capacity of hepatocytes but also the system of its control. PMID- 8233306 TI - [The behavior of embryonic nerve cells transplanted into the brain]. AB - Small solid pieces of 3H-thymidine labeled rat neocortex at day 20 of embryonic development were transplanted homotopically into the cortex of adult rats. Transplanted cells migrated from the site of implantation into cortical layers 2 4 of the host to the distance of about 1 mm. No labeled neurons was found in cortical layers 5-6 (the target site during their normal migration). Migration of glial cells was similar to that of astrocytes moving from the suspension grafts. In some cases, similar distribution of migrating donor cells around the implantation site was observed despite the fact that a solid grafted tissue was absent. PMID- 8233307 TI - [An analysis of chimeric mice obtained by the injection of the inner cell mass into the blastocyst]. AB - Mouse chimeras were produced using injections of ICM cells into blastocysts. Chimerism of resulting animals was determined by their coat color and spectrum of glucosephosphate isomerase isoenzymes. The use of modifications of the injection method for solving different genetic and embryological problems is discussed. PMID- 8233308 TI - [The morphology and postnatal development of the skin in purebred and mixed-breed cattle (Bos taurus)]. AB - Growth and differentiation of skin layers, hair follicles, and skin glands were studied in cattle of two genotypes during the period from early postnatal development until the age of 30 months. Development of skin and its derivatives was shown to proceed irregularly and include alternating periods characterized by an increase, temporary arrest, or even decrease of corresponding morphometric indices. On the basis of these data a conclusion is drawn that periodic changes in thickness of skin and its individual layers during ontogenesis are a natural phenomenon. It is suggested that a complex of morphometric indices can reflect specific characteristics of individual breeds and their productivity. PMID- 8233309 TI - [A comparative morphological analysis of the development of the ear in mammals]. AB - This paper is the first comparative study into prenatal ontogenesis of the peripheral auditory system in mammals. A wide range of ecologically different species was studied, including terrestrial, semi-aquatic and aquatic forms with auditory systems functioning at low, medium, and high frequencies. Structural peculiarities of the development of auditory organ were determined for each studied species in relation to its operation frequency and acoustic properties of the outer medium. The origin and individual stages of morphological adaptations found in semi-aquatic, aquatic, and echolocating species are described. The data obtained allowed to reveal general principles of development of the peripheral auditory system in mammals with different ecological specialization. PMID- 8233311 TI - Unions, health care, and the pansy-picking phenomenon revisited. PMID- 8233310 TI - [The survival of anlagen of the human hypothalamic supraoptic and paraventricular nuclei in the cavity of the third ventricle of the rat brain]. AB - We studied survival, integration into surrounding tissues, and morphogenesis of embryonic neurosecretory cells of human supraoptic and paraventricular hypothalamic nuclei transplanted into the third ventricle of the rat brain. It was found that embryonic brain cells survive after xenotransplantation and acquire features characteristic of corresponding cells during normal ontogenesis. These results suggest that the genetic program plays a key role in development of nervous cells. PMID- 8233312 TI - Lamellar flap corneal biopsy. AB - A modified technique of corneal biopsy is described that allows a lesion in the deep corneal stroma to be accessed for scraping or biopsy while sparing the stromal tissue anterior to the lesion. This technique may be especially valuable in cases of deep corneal pathology in which traditional methods of corneal biopsy risk excessive removal of tissue, which, in turn, can lead to complications such as thinning or perforation. PMID- 8233313 TI - Wedge resection for postkeratoplasty astigmatism. AB - Twelve patients underwent a modified wedge resection after keratoplasty; 11 of them were followed for 6 to 24 months. The average preoperative astigmatism was 15.40 diopters (range, 12.50 to 22.00 D). The wedge was resected from the donor side and the wound closed with 10-0 or 11-0 Mersilene sutures. Postoperatively, the average astigmatism was 4.36 D (range, 1.50 to 9.00 D), and the visual acuity, corrected with spectacles, was 20/40 or better in five cases (45%). Complications included high residual astigmatism or irregular astigmatism and extensive scarring due to the use of Mersilene sutures. Overall, modified resection of the wedge is an effective technique for managing postkeratoplasty high astigmatism. PMID- 8233314 TI - Focal and diffuse visual field defects: their relationship to intraocular pressure. AB - The view that visual field loss in "low-tension" glaucoma (LTG) is more focal than in ordinary or "high-tension" glaucoma (HTG) is controversial. The basis of the controversy may be the inappropriate presupposition that an arbitrary intraocular pressure (IOP) level effectively differentiates these entities. We attempted to determine whether meaningful differences exist between glaucoma patients classified on the basis of pattern of visual field loss rather than IOP. One hundred consecutive glaucomatous visual fields were classified as focal or diffuse by two independent observers masked to all other clinical information. Several clinical parameters were then evaluated, comparing patients with focal visual field loss, with those with diffuse loss. Maximal pretreatment IOP was significantly lower among the patients with focal visual field loss than it was among those with diffuse loss (21.5 mm Hg +/- 5.7 vs 25.9 mm Hg +/- 4.7; P < .01). There was no difference between the two groups in terms of age, blood pressure, visual acuity, or color vision. We conclude that focal glaucomatous visual field loss occurs at a lower IOP than diffuse loss and, as such, may be a marker by which to identify patients whose optic nerves are abnormally susceptible to glaucomatous injury. PMID- 8233315 TI - Transscleral neodymium:YAG laser cyclophotocoagulation for end-stage glaucoma, refractory glaucoma, and painful blind eyes. AB - We used transscleral Neodymium:YAG laser cyclophotocoagulation (TSYLCC) to treat 47 patients (47 eyes) with end-stage, refractory, or absolute glaucoma. The mean pretreatment intraocular pressure (IOP), 40.5 +/- 10.8 mm Hg, decreased a mean of 13 +/- 8 months following treatment to 15.6 +/- 10.6 mm Hg, a statistically significant change (P < .05). Our overall rate of success (final IOP < or = 25 mm Hg, with or without medications) was 79% (37 eyes). Early complications included pain (11 eyes) and inflammation (13 eyes); late complications included hypotony (7 eyes), high IOP (5 eyes), and decreased visual acuity (4 eyes). TSYLCC seems to be a safe, effective, and convenient outpatient procedure. PMID- 8233316 TI - Energy levels needed for cyclophotocoagulation: a comparison of transscleral contact cw-YAG and krypton lasers in the rabbit eye. AB - The energy levels needed for cyclophotocoagulation using either transscleral contact krypton or cw-YAG lasers were compared in pigmented rabbits. The same transscleral laser probe was used for both lasers. With the krypton laser, 0.25 J of energy caused macroscopically detectable lesions, and 1.5 J led to widespread destruction of the ciliary body. With the cw-YAG laser, the first detectable lesions were produced at 0.5 J. Similarly, at higher energies, twice as much energy was required using the cw-YAG as compared with using the krypton laser to produce comparable lesions. Histologically, lesions of the same macroscopic severity made with the two lasers were similar. We conclude that the transscleral contact krypton laser is an efficient instrument for cyclophotocoagulation. Although these results may not be directly applied to human eyes, it appears that the poorer scleral transmission of the krypton beam is offset by its higher level of absorption in the pigmented epithelium of the ciliary body. PMID- 8233317 TI - Pars plana vitrectomy for complications of retinal arterial macroaneurysms--a case series. AB - Retinal arterial macroaneurysms are distinct vascular lesions with a typically benign course. Laser treatment to these vascular lesions is controversial, and has been limited to cases of persistent macular edema. Devastating vision loss can occur, however, through vitreous or subretinal hemorrhage. We describe eight cases of retinal arterial macroaneurysms associated with subretinal or vitreous hemorrhage requiring pars plana vitrectomy. Final visual acuities ranged from 20/20 to 20/400 at most recent follow up. Until reliable indicators of rupture are identified, we suggest consideration of laser photocoagulation to clinically active, identifiable retinal arterial macroaneurysms within the macula. PMID- 8233318 TI - Perfluorocarbon liquid in traumatic vitreous hemorrhage and retinal detachment. AB - Patients with penetrating ocular trauma also may have severe vitreous hemorrhage and associated retinal detachment. Removing the hemorrhage and repairing the detachment can be a difficult surgical problem. Besides the limited surgical view due to the hemorrhage, an incomplete separation of the posterior hyaloid membrane can allow the detached retina to be drawn toward the port of the vitrectomy instrument, producing an inadvertent retinal tear. We have used perfluoroperhydrophenanthrene (Vitreon) to manage three cases of penetrating ocular trauma with concurrent retinal detachment and a partial vitreous detachment, either at the time of surgery or as noted ultrasonographically. The perfluorocarbon liquid helped to separate the partially detached posterior hyaloid membrane and flatten the detached retina. This maneuver simplified removal of the vitreous hemorrhage and management of the retinal detachment. PMID- 8233319 TI - Postoperative refraction and anterior chamber depth after complicated cataract surgery with vitreous loss and transscleral suture fixation of posterior chamber lenses. AB - The postoperative refraction and anterior chamber depth of 50 eyes that had complicated cataract extraction with vitreous loss were compared with those of 25 fellow eyes (that had uncomplicated cataract extraction) of the same patients. Transscleral suture fixation of the posterior chamber intraocular lens (PC-IOL) was required in 34 of the 50 study eyes; the lens was implanted onto the remaining defective capsule in the remaining 16. The anterior chamber depth was significantly deeper after transscleral suture fixation of the PC-IOL (mean, 0.54 mm; P < .01). The difference between the refraction of the eyes with transscleral suture fixation and that of the control eyes was +0.58 diopters. Although each surgeon should assess specific results and modify the lens calculations accordingly, we have found that with our technique, for eyes that have required transscleral suture fixation of a PC-IOL, +0.50 D should be added to the calculated IOL power. PMID- 8233320 TI - Acquired blepharoptosis secondary to essential blepharospasm. AB - We treated four patients with essential blepharospasm, receiving botulinum A toxin, in whom, although they had no preexisting blepharoptosis, a concurrent bilateral acquired blepharoptosis developed. Since the blepharoptosis did not improve after the period of time during which the effects of botulinum A toxin would have been expected to resolve (2 to 10 weeks), we judged that its development was unrelated to the toxin. We propose, rather, that the stretching, attenuation, disinsertion, or dehiscence of the upper eyelid levator muscle caused by the blepharospasm were at least partly responsible for the onset of the blepharoptosis. To ensure appropriate treatment in these cases, careful clinical evaluation is required to differentiate the two conditions. PMID- 8233321 TI - Subretinal hemorrhage after grid laser photocoagulation for idiopathic juxtafoveolar retinal telangiectasis. AB - A 56-year-old man had decreased vision secondary to bilateral idiopathic juxtafoveolar retinal telangiectasis. One month after grid laser photocoagulation for macular edema in the right eye, a subretinal hemorrhage developed. Over the next 2 months, the hemorrhage spontaneously resorbed and vision improved to 20/50. At 2-year follow up, without further treatment, vision was still 20/50. PMID- 8233322 TI - Suture use in pediatric cataract surgery: a survey. AB - Many different sutures have been advocated for use in closing the corneoscleral incision during cataract surgery. We surveyed 354 North American pediatric ophthalmologists to find out the type of suture they use for pediatric cataract surgery and any associated complications. Of the sutures indicated in the 244 (69%) surveys returned, 10-0 nylon sutures were the most popular choice despite a relatively large number of related complications and the occasional need for general anesthesia to remove them. PMID- 8233323 TI - Dynamic documentation of digital dacryocystography. AB - Digital dacryocystography has proved to be the optimum method of x-ray diagnostics in connection with managing obstructions of the lacrimal pathways. We present 50 patients in whom digital dacryocystography was performed with a computer-controlled x-ray unit and a C-arc coupled to an image intensifier TV system. A nonionic water soluble contrast medium was used. The bilateral contrast medium filling process appearing on the monitor was simultaneously recorded on videotape. Videocassettes, with patient data attached, were stored in a videotape library, providing repeatable diagnostic documentation and an excellent base for scientific analyses. PMID- 8233324 TI - Unilateral congenital cataract and binocular vision. PMID- 8233325 TI - Zeiss four-mirror gonioscopy. PMID- 8233326 TI - Reverse pupillary block in pigmentary glaucoma: follow up and new developments. PMID- 8233327 TI - Converting an American Optical ophthalmoscope handle to use standard C batteries. PMID- 8233328 TI - Diode versus Nd:YAG laser for cyclodestructive procedures. PMID- 8233329 TI - A modified removable suture in trabeculectomy. AB - We modified the closure of the scleral flap in trabeculectomy by using an externalized hemibow tie, easily removable in the early postoperative period. We used this externalized knot in 20 eyes of 17 patients undergoing trabeculectomy and in eight eyes of seven patients undergoing combined trabeculectomy and cataract extraction with intraocular lens implantation. The sutures were removed between 2 and 72 days after surgery (16.75 +/- 14.49 days). With a mean follow up of 11.36 months, the overall success rate was 78.6%. The sutures of nine eyes (32%) were removed due to elevated intraocular pressure and/or decreased bleb size. The immediate and long-term efficacies were 77% and 56%, respectively. The trabeculectomies we performed using this safe and simple technique, as compared with those using other methods of scleral flap closure, provided a lower incidence of postoperative complications due to overfiltration, and made it possible to increase the degree of filtration when needed during the early postoperative period. PMID- 8233330 TI - Association between surgically-induced astigmatism and cataract incision size in the early postoperative period. AB - One hundred sixteen eyes with cataracts were prospectively randomized into four surgical groups. Silicone and polymethylmethacrylate (PMMA) intraocular lenses were implanted into the posterior chamber of these eyes through 4-, 6-, or 11 millimeter incisions by one surgeon. Surgically-induced astigmatism was evaluated in terms of: (1) the proportion of eyes achieving a refractive cylinder of 1.5 or less, without regard to axis; and (2) the induced cylinder, with regard to axis, using vector analysis. One week postoperatively, the eyes with a 4-millimeter incision had a significantly lower mean induced cylinder than those with larger incisions (P = .019); and all three phacoemulsification groups had a significantly higher proportion of eyes with a cylinder of 1.5 or less than the nuclear expression group (P < .01). One month postoperatively, the 4-millimeter incision group had a significantly lower mean induced cylinder (P = .044) as well as a significantly higher proportion of eyes with a cylinder of 1.5 or less, than the larger-incision groups (P = .05). Three months postoperatively, no value of any parameter in any group differed significantly from that of that parameter in any other group. PMID- 8233331 TI - Use of perfluoroperhydrophenanthrene in the management of posteriorly dislocated crystalline and intraocular lenses. AB - We describe a series of 28 patients in whom a posteriorly dislocated crystalline or intraocular lens was successfully removed with the intraoperative use of perfluoroperhydrophenanthrene (Vitreon), a heavy liquid perfluorocarbon. The Vitreon was used to float the lens into the midvitreous cavity, thereby avoiding hazardous micromanipulation with intraocular instruments in the macular area. In addition, the Vitreon served to protect the posterior retina by keeping the posteriorly dislocated lens fragments from falling backward onto the posterior pole as the lens was being removed from the eye. Postoperative visual acuity improved in 73% of the patients, remained the same in 15%, and worsened in 12%. PMID- 8233332 TI - Deep venous thrombosis following immobilization after retinal detachment surgery. AB - Although deep venous thrombosis (DVT) and pulmonary embolism are potentially fatal complications of numerous surgical procedures, DVT associated with ophthalmic surgery has rarely been reported in the current literature. We report the development of a DVT in a patient following complicated vitreoretinal surgery. PMID- 8233333 TI - Silicone oil tamponade for retinal detachment and delayed treatment of retinal tears. AB - Four patients underwent a vitrectomy procedure for retinal detachment associated with anterior and posterior proliferative vitreoretinopathy (D-1 type). In each patient, an internal tamponade was achieved with 1000-centistoke silicone oil. Preretinal and/or subretinal hemorrhage that was difficult to remove during surgery, as well as remnants of subretinal fluid, prevented complete intraoperative treatment of the retinal tears or the retinotomy with endolaser photocoagulation. The silicone oil tamponade kept the retina flat until laser treatment was applied to the retinal tears several weeks later, when the retinal blood and subretinal fluid had reabsorbed. When circumstances make achieving intraoperative laser marks difficult, silicone oil tamponade appears to be superior to a long-lasting gas tamponade. It provides a more permanent tamponade than gas and excellent visualization of the fundus several weeks after surgery, at which time laser marks can be completed. PMID- 8233334 TI - Migration of solid silicone encircling element through four rectus muscles. AB - Anterior migration of an encircling element through a rectus muscle is an unusual complication of scleral buckling surgery. We describe two patients with migration of a solid silicone encircling element through the insertions of all four rectus muscles after retinal detachment surgery. Buckle migration was associated with elevated intraocular pressure and glaucomatous optic atrophy. Rectus muscle function was preserved in both patients. Despite transsection of the anterior ciliary blood supply, anterior segment ischemia did not develop. PMID- 8233335 TI - Pseudo-Brown's syndrome as a complication of glaucoma drainage implant surgery. AB - Two cases of pseudo-Brown's syndrome occurring after superior nasal implantation of glaucoma filtering devices are described. We hypothesize that this restriction in ocular motility was due to the mechanical effects of large filtering blebs or increased inflammation and scar formation in the superior nasal quadrant. Serial ultrasound examinations of our patients demonstrated development of a large bleb, coincidental with the disturbance in motility. Because of the risk of strabismus, we no longer recommend placing large implant devices in the superior nasal quadrant in eyes that have good vision. PMID- 8233336 TI - Prilocaine: an old anesthetic agent and a new ophthalmic procedure. AB - We present a new local anesthetic technique for ophthalmic surgery that reduces the risks associated with retrobulbar and peribulbar anesthesia. This method uses topical proparacaine plus 1.5 mL of prilocaine (3%) with felypressin injected into the subconjunctival (sub-Tenon's) space. Of 5210 consecutive adult patients in whom the technique was used, all demonstrated adequate analgesia. Sixty-three (1.2%) of the eyes required supplemental analgesia, provided by a single injection of prilocaine (0.5 mL). Ecchymosis and subconjunctival hemorrhage developed in 63 (1.2%) of the eyes. There were no instances of ptosis. PMID- 8233337 TI - Indications and technique for reopening closed filtering blebs using the Nd:YAG laser--a review and case series. AB - We reviewed the results of Nd:YAG laser treatment used with gonioscopy to reopen the internal sclerostomy in 10 consecutive patients with failing filtering blebs in whom the sclerostomy had become occluded internally with a membrane or iris after filtering surgery. Sixteen to 50 applications of 6- to 6.9-millijoules Nd:YAG laser irradiation were directed to the internal sclerostomy site. In 6 of the 10 patients, filtration was successfully reestablished, initially reducing the mean intraocular pressure (IOP) from 26.5 mm Hg (range, 20 to 40 mm Hg) to 8.6 mm Hg (range, 5 to 16 mm Hg), with an average IOP of 13.5 mm Hg at 7 months after the laser treatment. Nine of the patients were on significantly fewer glaucoma medications. We conclude that the YAG laser can be successfully used to reopen the fistula by the internal approach in carefully-selected patients who previously have had well-established filtering blebs. Identification of these suitable candidates involves careful gonioscopy. PMID- 8233338 TI - Repair of iridodialysis using a 17-millimeter straight needle. AB - A technique for repair of an iridodialysis using a 17-millimeter straight needle is described. Using this needle greatly facilitates the surgery and enables maintenance of a closed system, without loss of the anterior chamber or trauma to the cornea. PMID- 8233339 TI - Pars planitis presenting with vitreous hemorrhage. AB - A 31-year-old woman complained of a sudden, painless decrease in vision of the right eye. Ophthalmic examination demonstrated a vitreous hemorrhage in the right eye secondary to peripheral neovascularization. The left eye demonstrated findings consistent with pars planitis. Before this episode, vision was good in both eyes. Although rare, vitreous hemorrhage can occur in the setting of pars planitis and may be a presenting symptom. Pars planitis should be considered in the differential diagnosis of young patients with vitreous hemorrhage. PMID- 8233340 TI - Sodium hyaluronate dissection of conjunctival flap for trabeculectomy after extracapsular cataract extraction. PMID- 8233341 TI - Keratoplasty in the presence of anterior chamber drainage tube. PMID- 8233342 TI - Use of the infrared refractor to measure the pupillary diameter in darkness. PMID- 8233343 TI - Linguistic novelties in refractive surgery. PMID- 8233344 TI - Restoration of the outer blood-retinal barrier after krypton laser photocoagulation. AB - The restoration of the outer blood-retinal barrier following krypton laser injury to the rat retina was studied at small and large laser lesions using intravenously injected sodium fluorescein, horseradish peroxidase, and catalase. The regenerated choriocapillaris and new blood vessels were permeable to fluorescein and peroxidase, but not to catalase. The regenerating retinal pigment epithelium gradually reformed a continuous sheet of cells covering all small laser sites and the periphery of large lesions. Zonulae occludens between the regenerated cells restored the outer retinal barrier and prevented diffusion of peroxidase into the retina. This occurred along Bruch's membrane and the new blood vessels that were covered by the regenerated pigment epithelial cells, but not in the center of the large lesion that was not relined by regenerated cells. PMID- 8233345 TI - Experimental corneal cryopreservation: impact of postmortem time on corneal endothelial cell survival. AB - Clinical and experimental studies with rabbit and human corneas have shown the correlation between short postmortem times and successful corneal cryopreservation. In this experimental study we investigated this phenomenon considering the latent freeze-thaw-induced cell damage. Enucleated eye-balls of freshly slaughtered pigs were stored in moist chambers at 4 degrees C for 2, 4, 8, 16, 32, and 72 h before cryopreservation. After thawing, the corneas were organ-cultured for 1 day. After staining with trypan blue and alizarin red S the tissue was evaluated morphometrically, calculating the amount of necrotic areas on the central corneal surface and the endothelial cell density. Corneas stored up to 32 h before cryopreservation showed no difference regarding the amount of necrosis and endothelial cell density compared to freshly cryopreserved tissue. Corneas stored 72 h before cryopreservation revealed no endothelial cell survival. We conclude that a post-mortem time of up to 32 h before corneal cryopreservation has no influence on endothelial cell survival. PMID- 8233346 TI - Regional differences in the polypeptide composition of pig lens intracellular matrix and plasma membrane. AB - Analysis of pig lens urea-soluble fractions (USF) by gel filtration and SDS-PAGE revealed that the cortical USF is characterized by a high content of noncrystallin bands of 42, 44, 52, 55, 100 and 200 kD. These bands are markedly decreased in the nuclear USF though the 44-kD band to a lesser extent. These results suggested that pig lens nucleus contains much less cytoskeletal proteins than the cortex. Analysis of pig lens membrane showed that MP26 and MP17 are the major intrinsic membrane proteins and the relative amount of alpha A and beta B1b chains attached to the membrane are more abundant in the nucleus than in the cortex. PMID- 8233347 TI - Distribution of autofluorescence in the rabbit corneal epithelium. AB - Autofluorescence from reduced pyridine nucleotides (PN) and oxidized flavoproteins (Fp) was measured in order to detect the difference in redox states in rabbit corneal epithelium. The enucleated rabbit eye was mounted in an eye bank eye container with McCarey-Kaufman medium, and the autofluorescence was measured using ocular redox fluorometry as a function of depth. The PN signal distributed evenly whereas the Fp signal was greater in the posterior epithelial region than in the anterior region (p < 0.05). The PN/Fp ratio, a sensitive indicator of tissue redox state, was less in the posterior region. After the application of 1 mM of potassium cyanide in the medium, the ratio increased significantly in each layer (p < 0.001), and the difference between anterior and posterior region diminished. These results indicate that ocular redox fluorometry has the potential to resolve the redox states of the various layers of the corneal epithelium. The posterior region of the epithelium is more active in mitochondrial respiration than the anterior region. PMID- 8233348 TI - Human retinal pigment epithelium in vitro: influence of low oxygen tension, glucose and insulin. AB - Retinal pigment epithelium (RPE) cells migrating through the damaged retina into the vitreous body seem to play an important role for the pathogenesis of proliferative vitreoretinopathy (PVR) and other proliferative retina diseases. It is so far not known how the RPE cells are able to survive in the vitreous body without contact to the blood vessels of the choroid supplying them with oxygen and nutritive substances. To answer this question, we studied growth characteristics and sensitivity to glucose and insulin of human RPE cells, incubated with reduced oxygen partial pressure. In the first study, RPE cultures of 58 postmortem human eyes were grown with 5% O2/5% CO2 and with standard conditions (20% O2/5% CO2). The growth was assessed in five graded stages. Our data show that human RPE grows better under 5% oxygen than under 20% O2 (p < 0.0001). In consideration of this effect, we cultivated, in a further study, pigment epithelium of 49 postmortem human eyes with 5% oxygen and with 4 different glucose concentrations with and without addition of insulin. We found that glucose in higher concentrations was a potent stimulator of growth, whereas insulin was a modest stimulator when used alone. The combination of glucose and insulin was significantly more effective (p = 0.01) in the period of the first 7 days. These results suggest that proliferation of human RPE cells can be increased by oxygen reduction, glucose and insulin. These interactions may help in understanding the pathophysiology of retina damage and proliferative retina diseases like PVR. PMID- 8233349 TI - Healing response of the epithelium after lateral needle injury of mouse lens. AB - Characteristic changes of the epithelial healing process in germinative and equatorial zones of the ddY mouse lens were studied morphologically. After germinative zone injury, the wound was covered by proliferating epithelial cells which increased and migrated posteriorly as a function of time after injury. Epithelial posterior migration from the lens equator was mild. By contrast, with equatorial injury, the wound was covered by proliferated epithelial cells. Moreover, the equatorial area was characterized by disconfiguration of the lens bow and posterior epithelial migration. The results suggest that the larger capsulectomy in IOL operation, which damages the peripheral part of the lens epithelium, might contribute to secondary cataract. PMID- 8233350 TI - Comparison of electro-oculogram recording methods. AB - The purpose of this study was to compare the variability of the amplitude ratios and latencies of the electro-oculogram (EOG) light peak (LP) in three different recording conditions: Arden ratio (dark trough (DT): 12 min), modified Arden ratio (DT: 15 min) and light peak/dark baseline ratio (LP/BL: 35 min of dark adaptation). Additionally, EOGs from eyes with dilated and undilated pupils were recorded. The light intensity stimulating the eyes with dilated pupils was attenuated 1 log unit. The EOG amplitude ratios displayed no significant difference between the three conditions tested. The comparison of implicit times (time from onset of light exposure to LP) revealed significantly (p < 0.001) shorter values when the recording of the LP was preceded by a dark adaptation of 35 min. The broadening of the curves in DT/LP recordings might be caused by a superposition of a residual dark oscillation on the LP. The recordings from eyes with dilated pupils--stimulated with 1 log unit lower intensity--revealed lower amplitude ratios than the recordings from eyes with undilated pupils. PMID- 8233351 TI - Role of hydrogen peroxide in experimental optic neuritis. A serial quantitative ultrastructural study. AB - In order to determine the role of H2O2 in demyelination of the optic nerve, serial quantitative analysis of H2O2-derived cerium perhydroxide reaction product particles was obtained by computerized digitization of electron micrographs of the myelinated retrobulbar optic nerve, unmyelinated optic nerve head, and the optic nerve sheath of guinea pigs sensitized for experimental allergic encephalomyelitis (EAE) and euthanized 3-14 days later. We found that cerium perhydroxide reaction product particles were greatest in the myelinated optic nerve 3 days after antigenic sensitization, but at this focus decreased 7-14 days after antigenic sensitization. Reaction product accumulated in the unmyelinated optic nerve head and optic nerve sheath 3-14 days after sensitization. These results in the myelinated optic nerve suggest H2O2 consumption results in peroxidation of myelin lipid as demyelination proceeds 7-14 days after antigenic sensitization. Hydrogen peroxide accumulation in the optic nerve head and the optic nerve sheath appears to provide a reservoir for diffusion of H2O2 into the retrobulbar optic nerve and adjacent perineural nerve, contributing to the frequent predilection for optic nerve involvement in EAE and perhaps in multiple sclerosis. PMID- 8233352 TI - Angle closure glaucoma in Alagille syndrome. A case report. AB - Alagille's syndrome or arteriohepatic dysplasia is a unique and uncommon congenital disorder exhibiting a combination of systemic and ocular abnormalities. It is of interest to the ophthalmologist presenting with a combination of anterior and posterior segment anomalies and it is unique for the paediatrician in that these anomalies distinguish this condition from all other causes of neonatal jaundice. The authors describe a case of a young girl with Alagille's syndrome who developed bilateral acute angle closure, a hitherto unreported finding in this condition. PMID- 8233353 TI - Eighties and nineties have witnessed an explosion of new information on the genetics of human diseases. PMID- 8233354 TI - Olivopontocerebellar atrophy with retinal degeneration. Fundus characteristics and diagnostic MRI findings. AB - Olivopontocerebellar atrophy with retinal degeneration (OPCA type III) and autosomal dominant cerebellar atrophy of late onset (type II) appear to represent the same disease which is characterized by dominantly inherited cerebellar ataxia and pigmentary retinal degeneration. In June, 1988, a 15-year-old girl presented with objective visual acuity loss but no other findings. When seen again in January, 1991, macular changes were noted. At this time, her mother, diagnosed with multiple sclerosis, was examined and found to have atrophic macular lesions and poor vision. A brain MRI was performed which showed findings diagnostic of OPCA type III, including cerebellar and pontine atrophy and specific loss of the inferior olives. Fundus features, electroretinographic results and other clinical findings in OPCA type III are discussed and diagnostic MRI lesions are demonstrated. PMID- 8233355 TI - Non-conventional ultrasonography (power spectrum analysis) in the management of retinoblastoma. AB - The differential diagnosis among the diseases that may simulate retinoblastoma is supported by several techniques; however, none of them can give an undeniable answer. In the authors' opinion the tissue characterization by power spectrum analysis of the radiofrequency ultrasound data may play an important role in the backscattered signal spectrum while being sensitive to the spectral shift trend. In order to evaluate the patterns of regression of quiescence of retinoblastoma foci after conservative treatment the authors try to create a model of tissue characterization that provides information not available in conventional A & B scan ultrasonography about cell type, vascularization and necrosis. PMID- 8233356 TI - Did atomic bomb radiation influence the incidence of retinoblastoma in Nagasaki and Hiroshima? AB - The authors studied the effect of atomic bomb radiation in Nagasaki and Hiroshima on the incidence of retinoblastoma by examining the history of exposure to atomic bomb radiation among the grandparents and parents of retinoblastoma patients in Nagasaki and Hiroshima. Of 85 patients born in Nagasaki Prefecture from 1965 to 1986 and in Hiroshima Prefecture from 1966 to 1986, 42 had records to whether or not their grandparents and parents had been exposed to atomic bomb radiation. Seven had parents or grandparents exposed to atomic bomb radiation. These patients showed neither a high incidence of bilateral retinoblastoma nor a family history of retinoblastoma, although a higher incidence of cancer in the family history was found in the exposed group. Exposure to radiation showed no correlation with the onset of retinoblastoma. Moreover, the incidence of retinoblastoma in Nagasaki and Hiroshima Prefectures was almost the same as the average incidence in Japan. The authors failed to find evidence of increased incidence of retinoblastoma in the offspring of those exposed to atomic bomb radiation in Nagasaki and Hiroshima. However, this does not mean that the effect of radiation on the occurrence of retinoblastoma can be ruled out. PMID- 8233357 TI - Cataracts in glucose-6-phosphate dehydrogenase deficiency. AB - Glucose-6-phosphate dehydrogenase (G-6-PD) deficiency plays an important biochemical role in the metabolism of the lens. Controversies exist in the literature on the possible association between G-6-PD deficiency and the development of cataracts. The authors present ten patients, aged between infancy and 40 years of age, who were admitted for bilateral congenital or presenile cataracts. These patients had no ocular or systemic disease which might have caused their cataract. The only systemic finding they had was G-6-PD deficiency. Two other patients among the families described suffered from bilateral congenital or presenile cataracts with no G-6-PD deficiency. This deficiency state does not appear to play a role in the production of their cataracts. PMID- 8233358 TI - Cataracts in Morquio syndrome (mucopolysaccharidosis IV A). AB - Three siblings with Morquio syndrome (mucopolysaccharidosis IV A) are described. In addition to the characteristic dwarfism with skeletal deformities, odontoid anomalies, hearing loss and corneal clouding, the authors found almost identical lens opacities in all three patients. Lenticular opacities have not previously been described in patients with Morquio's syndrome IV A. PMID- 8233359 TI - Anterior segment malformations in 18q- (de Grouchy) syndrome. AB - The authors examined two patients with deletions of the terminal end of the long arm of chromosome 18. The ocular findings in one patient with 46, XX, del 18 (q21) consisted of hypertelorism, epicanthus, strabismus, myopia, microphthalmia, microcornea, corneal opacities, iris hypoplasia with full thickness defects, corectopia and large peripapillary staphylomata. The second patient with 46, XX, del (18) (pter --> q21.2 :: q22 --> qter), inv (21) (q21 --> p12 :: q21 --> qter) only had epicanthus, strabismus, myopia and peripapillary crescents. Based on the findings in these two patients and on a review of previously reported patients with del 18 qter it appears that the loss of band 18q23 may be responsible for malformations of the anterior segment in the 18q-syndrome. PMID- 8233360 TI - Acetazolamide in dominant cystoid macular dystrophy. A pilot study. AB - Eight patients (four men, four women), with low visual acuity caused by autosomal dominant cystoid macular dystrophy, were treated daily with oral 250 mg dose acetazolamide. Treatment ranged from two to 17 months. None of these eight patients had improvement of visual acuity of more than 0.1. CONCLUSION: Treatment with 250 mg acetazolamide appears not to be an effective therapy for cystoid macular oedema in dominant cystoid macular dystrophy. The electroretinography b wave/a-wave ratio was normal. The primary lesion in dominant cystoid macular dystrophy remains obscure. PMID- 8233361 TI - Integrating public health in professional education. AB - In view of the spiralling costs of treating those who are ill, the author urges greater efforts toward disease prevention. Integration of public health education and professional training is advocated. The curriculum of the School of Public Health, University at Albany, is described. PMID- 8233362 TI - The critical importance of epidemiology in public health and professional education. AB - The author discusses the uses of epidemiology in identifying underserved populations and the positive impact of preventive health care on poverty and social distress. The U.S. Public Health Service document, Healthy People 2000, and its recommendations regarding eye care, are discussed. PMID- 8233363 TI - Manpower issues affecting public health policy. AB - This paper addresses the efficiency of health care delivery in several countries. Confusions arising from comparing the share of gross domestic product going to health care in various countries are discussed, along with the varying levels of coverage enjoyed by the populace. Factors contributing to the rising cost of health care are analyzed and suggestions for improvement are offered. PMID- 8233364 TI - The making of health policy affecting health professions. PMID- 8233365 TI - Theoretical effect of refractive error and accommodation on longitudinal chromatic aberration of the human eye. AB - Simple formulas based on reduced eyes have been developed to predict the variation in longitudinal chromatic aberration with variation in ametropia or accommodation. Two formulas were developed, one for axial ametropia and one for refractive ametropia. The latter also served as a model for accommodation. The results using the formulas are in close agreement with results obtained using raytracing through more sophisticated models. Combining the results of different methods gives the following predictions of change in chromatic difference of focus, between wavelengths of 400 and 700 nm, with change in each diopter of refractive error or accommodation: axial ametropia 0.012 to 0.017 D (0.6 to 0.9%), refractive ametropia 0.05 D (2.2 to 2.4%), and accommodation 0.04 to 0.05 D (2.1 to 2.6%). The chromatic aberration effects of correcting lenses with low dispersion are intermediate in effect and opposite in sign to the effects of corresponding degrees of axial ametropia and refractive ametropia. PMID- 8233366 TI - Distance visual acuity and monovision. AB - We have investigated aspects of visual acuity with monovision correction. Binocular distance visual acuity with monovision was approximately equal to monocular visual acuity for addition powers (monocular defocus) of +1.00 to +2.50 D, using both high and low contrast logMAR visual acuity charts. Neither the eye chosen to be defocused (dominant or nondominant) or pupil size (3.5, 5 and 7 mm) affected the binocular visual acuity loss with monovision for high contrast charts. We also investigated the effect of induced residual astigmatism (+0.50, +1.00, and -1.00 D) on binocular distance visual acuity in monovision correction. Residual astigmatism caused a significantly greater reduction of binocular visual acuity in the monovision condition than it did in normal binocular conditions. This effect appears to be related to a process of meridional interocular suppression. It may therefore be of clinical importance to correct low amounts of residual astigmatism in monovision corrections to provide optimum binocular visual acuity. PMID- 8233367 TI - Intraocular pressure response to scleral indentation. AB - PURPOSE: Intraocular pressure (IOP) is influenced by many factors. Our purpose was to determine the immediate response of the IOP to scleral indentation (SI). METHODS: Twenty-five subjects had IOP measured using Goldmann applanation tonometry before and after SI. The total indentation time was 40 s. IOP was remeasured in a masked fashion. RESULTS: Average IOP for the control eyes was 15.32 mm Hg (range: 10 to 22 mm Hg) before SI and 14.80 mm Hg (range: 9 to 24 mm Hg) after SI. The average IOP for the subject eyes was 15.08 mm Hg (range: 8 to 22 mm Hg) before SI and 13.14 mm Hg (range: 6 to 22 mm Hg) after SI. CONCLUSIONS: These results indicate an average decrease of 1.94 mm Hg for the subject eyes vs. 0.52 mm Hg for the control eyes in response to SI. This represents a statistically significant drop in IOP among the subject eyes in response to SI (p < 0.001). PMID- 8233368 TI - Rigid contact lens design equivalencies of overall diameter and axial edge lift: individual variations. AB - We compared the effects on rigid contact lens tear pump efficiency of changes in lens overall and optic zone diameter and of changes in lens axial edge lift by measuring corneal oxygen uptake on the right eyes of six human subjects after 5 min of both static and dynamic contact lens wear. Two sets of five lenses were worn by each subject. In set 1, overall diameter varied from 8.2 to 9.4 mm in 0.3 mm steps, with the optic zone diameter being 1.4 mm smaller and a 0.09 mm axial edge lift. In set 2, axial edge lift varied from 0.05 to 0.13 mm in 0.02-mm steps; overall diameter was 8.8 mm, and optic zone diameter was 7.4 mm. For the pooled data, a 0.3-mm decrease in overall diameter was approximately equivalent to a 0.03-mm increase in axial edge lift as a means of enhancing tear pump efficiency. However, considerable intersubject differences were exhibited, with the following increases in axial edge lift being equivalent to the 0.3-mm reduction in overall diameter: 0.01, 0.01, 0.04, 0.06, 0.09, and > 0.20 mm. This illustrates that subject factors (i.e., palpebral aperture size, lid-to-cornea geometry, corneal toricity, peripheral corneal topography, lid tension) influence lens positioning, movement, and tear pump efficiency. PMID- 8233369 TI - Effects of ultraviolet radiation on contact lens parameters. AB - Ultraviolet (UV) radiation has been proposed as an alternative method for contact lens disinfection. In order for UV radiation to be considered a viable contact lens disinfection method, its effects on lens polymers and parameters must be minimal. To evaluate this, soft contact lenses from each of the four FDA categories, soft lenses with handling tints, and rigid gas permeable (RGP) lenses were exposed to UV radiation in a controlled laboratory setting and their parameters measured. After 20 h of UV exposure, we found statistically significant changes in at least one parameter for each lens type evaluated. Most of these changes were small and within the error of measurement and, therefore, were not considered clinically significant. However, the group 2 soft lenses showed an increase in all four measured parameters--power, center thickness, diameter, and water content. These changes may have an impact on lens fitting characteristics and performance. Other than these changes for group 2 soft lenses, 20 h of UV exposure appeared to have little clinical effect on the contact lenses evaluated. This suggests that UV radiation may prove to be a feasible alternative method of disinfection for most types of contact lenses. PMID- 8233370 TI - Electroretinogram in unilateral vascular stress in nondiabetic and diabetic subjects. AB - PURPOSE: The oscillatory potentials (OP's) of the flash-elicited electroretinogram (ERG) are vulnerable to changes in retinal circulation such as occur in insulin-dependent diabetes mellitus (IDDM). In an earlier study it was shown that rod-isolated ERG's were more sensitive than cone-isolated ERG's to transient changes in the ocular perfusion pressure (OPP). In the present study, a comparison was made of the susceptibility of scotopic white, red, and blue flash OP's to transiently decreased perfusion in normals. Scotopic white flash OP's were then compared to those obtained in subjects with IDDM. METHODS: The OPP was decreased by 20 and 40% through scleral suction in 10 normotensive healthy volunteers, as well as in 11 young subjects with IDDM. RESULTS: The data indicate that white flash OP's include early cone and later rod contributions. Both red and blue OP's were more variable than white OP's. For white flash OP's, component specific changes to decreased OPP were found. CONCLUSION: The overall OP response profiles for the diabetic subjects during a stepwise short-term decrease in the OPP did not differ from those for nondiabetic subjects. The results are discussed in relation to retinal vascular autoregulation (AR) and choroidal blood flow. PMID- 8233371 TI - Initial cross-sectional results from the Orinda Longitudinal Study of Myopia. AB - BACKGROUND: Although investigations of human refractive error development and normal ocular growth have been conducted for the last 50 years, no previous study of refractive error and the ocular components has measured all the ocular components. METHODS: The Orinda Longitudinal Study of Myopia was initiated to characterize the development of refractive error and normal eye growth in a sample of predominantly Caucasian children ages 6 to 14 years. RESULTS: Cross sectional results from 530 children ages 5 to 12 years in the 1st, 3rd, and 6th grades are presented. CONCLUSIONS: This sample's refractive error decreased toward emmetropia with age from an average of +0.73 D at age 6 years to an average of +0.50 D by age 12 years. Between the ages of 6 and 12 years, the vitreous chamber elongated (by 0.52 mm) and the crystalline lens power decreased (by 1.35 D); surprisingly, the crystalline lens thinned by 0.14 mm during this same time period. PMID- 8233372 TI - Assessment of the reliability of calculations of the coefficient of variation for normal and polymegethous human corneal endothelium. AB - In endothelial morphometry, uncertainty exists concerning how many cells should be measured. A study was undertaken to calculate mean cell area and coefficient of variation (COV) of cell areas using different numbers of cells from photo slitlamp pictures and published micrographs. Groups of 65, 95, or 165 tesselated cells were measured and area and COV values calculated in progressive sets of 5 cells; each pair of values was compared to that obtained using all cells in each group. The results show that, for both normal (homomegethous) and irregular (polymegethous) endothelia, even cell counts as low as 50 cells can usually provide average cell area values that are within 1 to 2% of the values estimated from larger groups of cells. A similar reliability was observed for estimates of COV for normal endothelia. However, for polymegethous endothelia, even with 100 cells analyzed, the estimates of COV generally only approached a +/- 4% reliability. This uncertainty in COV estimates should be considered in both comparative studies and in regression analyses of COV changes over time or other variables. PMID- 8233373 TI - Diagnosis and management of temporal arteritis: a review and case report. AB - Temporal arteritis is an insidious disease which, if not recognized and treated with high-dosage oral prednisone or intravenous prednisolone, can result in unilateral or even total blindness due to anterior ischemic optic neuropathy (AION) or closure of the central artery of the retina. Unfortunately, the symptoms and clinical signs of temporal arteritis mimic those of a number of other conditions including angle-closure glaucoma, hypertension, migraine, trigeminal neuralgia, temporomandibular joint syndrome, carotid artery occlusive disease, Foster-Kennedy syndrome, and nonarteritic AION. When a patient complains of a severe pain in the temporal region, along with scalp tenderness and a feeling of malaise or depression--with or without episodes of transient loss of vision--he or she should be referred for a diagnostic work-up which includes an erythrocyte sedimentation rate and a temporal artery biopsy. We present here a review of the recent literature concerning temporal arteritis, followed by a report of an unusual case in which high-dosage prednisone therapy was effective in relieving the patient's symptoms and lowering the sedimentation rate in spite of a negative temporal artery biopsy. PMID- 8233374 TI - Cataract surgery in children with JRA. PMID- 8233375 TI - Vitrectomy specimen culture technique. PMID- 8233376 TI - Precortical vitreous pockets and proliferative retinopathy. PMID- 8233377 TI - Treatment of Propionibacterium endophthalmitis. PMID- 8233378 TI - Cost of sustained release treatment of CMV retinitis. PMID- 8233379 TI - Corneal trabeculectomy. PMID- 8233380 TI - Retinal pigment epitheliopathy after macular hole surgery. PMID- 8233381 TI - Excimer PRK for myopia. PMID- 8233382 TI - Surgical management of idiopathic macular holes. PMID- 8233383 TI - Relationship of drinking alcohol and smoking to prevalence of open-angle glaucoma. The Beaver Dam Eye Study. AB - PURPOSE: To evaluate a possible association of glaucoma with alcohol ingestion and cigarette-smoking behavior. METHODS: A population-based survey was performed of persons 43 to 84 years of age in Beaver Dam, Wisconsin (n = 4926). The diagnosis of glaucoma was based on visual field testing, measurement of intraocular pressure, and cup-to-disc ratios based on detailed protocols for examination, measurement, and gradings of photographs. Alcohol intake and smoking history were based on questionnaire responses. The prevalence of glaucoma in "heavy drinkers," whether current or past, was contrasted with the prevalence in those not reporting heavy drinking. Prevalences in never, past, and current smokers were compared. RESULTS: There was no difference in frequency of glaucoma by drinking status. Similarly, there was no difference in the frequency of glaucoma by cigarette-smoking status. CONCLUSION: In these prevalence data, neither heavy drinking nor cigarette-smoking behavior was related to the prevalence of open-angle glaucoma. This finding may reflect a real absence of a pathologic effect. PMID- 8233384 TI - Long-term follow-up of diode laser trabeculoplasty for primary open-angle glaucoma and ocular hypertension. AB - BACKGROUND: Initial studies of laser trabeculoplasty using infrared energy (810 nm) emitted by diode semi-conductor lasers have been encouraging. A 2-year study of diode laser trabeculoplasty (DLT) in the control of primary open-angle glaucoma (POAG) and ocular hypertension has been completed. METHODS: Patients with uncontrolled POAG or ocular hypertension were treated with DLT to one half of the trabecular meshwork using a trabeculoplasty lens. Spot size was 100 microns, exposure time was 0.2 second, and mean power was 1096 mW (+/- 46.5 mW). The desired endpoint was a mild blanching of the meshwork only. RESULTS: Twenty five eyes of 16 patients were treated. Mean intraocular pressure reduction was 9.24 mmHg (+/- 3.4 mmHg) at 6 weeks, 9.32 mmHg (+/- 3.6 mmHg) at 3 months, 9.34 mmHg (+/- 3.8 mmHg) at 6 months, 8.42 mmHg (+/- 2.62 mmHg) at 12 months, 8.14 mmHg (+/- 3.42 mmHg) at 18 months, and 7.9 mmHg (+/- 3.63 mmHg) at 24 months. No pressure peaks (> 5 mmHg) were recorded after therapy. Inflammation and discomfort were minimal after laser treatment. Of 16 eyes examined at 2 years, there were no peripheral anterior synechiae. During the course of the study, six eyes became uncontrolled, despite one session of DLT. Four eyes regained control with a further session of DLT, but two required trabeculectomy. CONCLUSION: Diode laser trabeculoplasty is an effective form of therapy in POAG and ocular hypertension. Hypotensive effects and success rates are comparable with argon laser trabeculoplasty (ALT). Reduced inflammation after laser treatment may be due to reduced absorption of infrared energy by the melanin of the anterior segment. The portable nature of these lasers may allow for laser delivery in developing countries and remote situations. PMID- 8233385 TI - Safety and efficacy of mitomycin C trabeculectomy in southern India. A short-term pilot study. AB - PURPOSE: Glaucoma is a major cause of blindness in developing nations. Medical and laser therapy have limitations in this environment. Filtration surgery alone often fails in young, darkly pigmented, individuals. The authors, therefore, evaluated the safety and efficacy of filtration surgery, using topical mitomycin C, in patients undergoing trabeculectomy in India. METHODS: A short-term pilot study used mitomycin C at the time of trabeculectomy in a population of southern Indians with various forms of glaucoma. A single 5-minute intraoperative application of 0.4 mg/ml mitomycin C was given to 76 consecutive patients requiring filtration surgery for the control of intraocular pressure (IOP). All patients were admitted for 6 days and were seen at 6-week intervals for a minimum of 18 weeks. RESULTS: The mean preoperative IOP was 35.3 +/- 9.6 mmHg. Postoperative IOP control was attained in 93.4% of patients as defined by an IOP of 20 mmHg or less without additional glaucoma medications (or an IOP reduction > 25% if the preoperative IOP was < or = 20 mmHg). No corneal epithelial defects or wound leaks were seen. The major complications were cataract formation and a decreased visual acuity in ten (13%) eyes. CONCLUSION: Mitomycin C-aided trabeculectomy was highly successful in controlling IOP in this darkly pigmented population and may become an important adjunct in the treatment of glaucoma in developing countries. PMID- 8233386 TI - Low-dose and high-dose mitomycin trabeculectomy as an initial surgery in primary open-angle glaucoma. AB - PURPOSE: The purpose of the study is to determine the optimum regimen of intraoperative administration of mitomycin as an adjunct to trabeculectomy. METHODS: Of 11 patients with primary open-angle glaucoma, 22 eyes that had not undergone any surgical intervention were included. In each patient, one eye was randomly allocated to a mitomycin 0.2-mg group and the fellow eye to a mitomycin 0.02-mg group. Mitomycin was applied for 5 minutes only once during trabeculectomy. The follow-up period was 6 to 17 months. RESULTS: Eleven (100%) eyes in the 0.2-mg group and 7 (63.6%) in the 0.02-mg group achieved successful control of intraocular pressure with or without topical antiglaucoma medication. Transient hypotony maculopathy (18%) and cataract progression (18%) were noted in the 0.2-mg group exclusively. The incidence of other complications was similar between the two groups. CONCLUSION: These data suggest that the most appropriate dose of mitomycin for primary surgery seems to be in between the two doses tested in the current study. PMID- 8233387 TI - Postoperative intraocular pressure abnormalities in the Silicone Study. Silicone Study Report 4. AB - BACKGROUND: Chronically abnormal intraocular pressure (IOP) may follow surgery for proliferative vitreoretinopathy (PVR), using either long-acting gas or silicone oil tamponade. Its prevalence and clinical significance are unclear. METHODS: In the Silicone Study, 241 eyes with severe (> or = C-3) PVR were treated with vitrectomy, randomized to perfluoropropane gas (C3F8) or silicone oil, and followed for 6 months or longer. Chronic IOP abnormalities, based on findings at two consecutive or any three postoperative visits, were defined as (1) low IOP (hypotony), 5 mmHg or less, or (2) elevated IOP, more than 25 mmHg. RESULTS: Eleven (5%) eyes had chronically elevated IOP and 58 (24%) had chronic hypotony. Chronically elevated IOP was more prevalent in eyes randomized to silicone oil than in those randomized to C3F8 gas (8% versus 2%; P < 0.05). Chronic hypotony was (1) more prevalent in eyes randomized to C3F8 gas than in those randomized to silicone oil (31% versus 18%; P < 0.05); (2) more prevalent in eyes with anatomic failure (48% versus 16%; P < 0.01); and (3) correlated with poor postoperative vision (P < 0.0001), corneal opacity (P < 0.001), and retinal detachment (P < 0.001). Factors prognostic of chronic hyotony included preoperative hypotony (P < 0.01), diffuse contraction of the retina anterior to the equator (P < 0.01), rubeosis (P = 0.02), and large retinal breaks (P = 0.02). In a multivariate analysis, diffuse contraction of the retina anterior to the equator remained an independent factor prognostic of chronic hypotony (odds ratio = 4.2), regardless of whether the retina was attached postoperatively. CONCLUSION: Intraocular pressure abnormalities are a common postoperative complication in eyes with PVR, and may occur with either C3F8 gas or with silicone oil. The presence of diffuse contraction of the retina anterior to the equator should alert the vitrectomy surgeon that the eye is likely to be hypotonus postoperatively. PMID- 8233388 TI - Flexible open-loop anterior chamber intraocular lens implants. AB - PURPOSE: To determine the efficacy of flexible open-loop anterior chamber intraocular lens (AC IOL) implants in cataract surgery complicated by vitreous loss and in secondary implantation. METHOD: Results were reviewed in 18 eyes in the former group and in 43 eyes in the latter group that were operated on by the author over an 8-year period. RESULTS: Of 18 eyes in the former group, 13 (72%) had a final visual acuity of 20/40 or better. Cystoid macular edema was the most common cause of the decrease in visual acuity. Of 43 eyes in the latter group, 37 (86%) had a final vision equal to the best-corrected preoperative vision. A retinal complication developed in 6 of 18 eyes that received a primary AC IOL with vitreous loss, whereas no retinal complications developed in any of the 14 eyes that received a secondary AC IOL with vitreous loss (P = 0.021). For secondary AC IOL implantation, eyes with vitreous loss treated by Weck-cel sponge vitrectomy had the same visual results as eyes without vitreous loss. CONCLUSION: Flexible open-loop AC IOLs are safe and effective for primary implantation after vitreous loss and for secondary implantation. PMID- 8233389 TI - Intravitreal pharmacokinetics of liposome-encapsulated amikacin in a rabbit model. AB - BACKGROUND: Intravitreal injection of antibiotics has become a standard therapy for bacterial endophthalmitis. The duration of effective antimicrobial levels in the vitreous after single injection, however, may not be long enough to get optimal response. The authors prepared liposome-encapsulated amikacin for prolonging the duration of intravitreal therapeutic concentrations and investigated the intravitreal pharmacokinetics of the liposomes and amikacin in phosphate buffer solution (PBS) as control. METHODS: The liposome-encapsulated amikacin was prepared by reverse-phase evaporation method. The intravitreal pharmacokinetics of the liposomes was compared with amikacin in PBS by fluorescence polarization immunoassay. Albino rabbits were randomly distributed into 12 groups. Rabbits in groups 1 to 6 and in groups I to VI (control groups) received an intravitreal injection of the liposome-encapsulated amikacin and amikacin in PBS, respectively. RESULTS: The encapsulation rate of amikacin was 91%. The time of 50% spontaneous degradation (half-life) of the liposomes in PBS (38 degrees C, pH 7.4) was 47.6 days, and the time of 50% release (half-life) of the drug from the liposomes in PBS was 84.8 hours. The vitreous amikacin concentrations in groups 1 to 6 were significantly greater (P < 0.05) than those in control groups I to VI in every time interval, except in groups 1 to 3 at 1 hour after injection. The difference was particularly obvious in the endophthalmitis groups. The clearance of encapsulated amikacin in vitreous appeared to be related to the state of blood-ocular barrier and to the structural integrity of vitreous. The distribution, the absorption, and the elimination of encapsulated amikacin in vitreous showed the first-order kinetics. CONCLUSION: The liposome-encapsulated amikacin prolonged half-life of the drug in vitreous. The results of the pharmacokinetic analysis suggested that in endophthalmitis, especially in severe cases, the liposomes may be preferable to conventional preparation. PMID- 8233390 TI - Retinal and other manifestations of incontinentia pigmenti (Bloch-Sulzberger syndrome). AB - BACKGROUND: One of the largest series of patients with incontinentia pigmenti who have undergone detailed ophthalmologic examination is reported here, including previously unrecognized findings with visually disabling consequences. METHODS: Thirteen females with incontinentia pigmenti from eight unrelated families were evaluated. The diagnosis of incontinentia pigmenti had been established previously by a referring pediatrician or dermatologist in 12 patients and by the authors in one other patient on the basis of retinal findings and history of characteristic skin manifestations. RESULTS: Abnormalities of the eye, central nervous system, skeleton, teeth, and hair develop in a majority of patients. The authors describe the evolution of retinal vascular abnormalities, and, for the first time, document foveal hypoplasia (in 4 of their 13 patients). The authors also provide the first report of a child with a normal brain shown on computed tomographic scan at 3 days old, which evolved to devastating cerebral ischemia, edema, and cortical blindness beginning at 6 days old. CONCLUSIONS: Incontinentia pigmenti should be included in the differential diagnosis of patients with peripheral retinal vascular nonperfusion, preretinal neovascularization, infantile retinal detachment, or foveal hypoplasia, particularly if there is evidence of characteristic dermatologic or other systemic manifestations. In infants with incontinentia pigmenti, retinal vascular anomalies are best detected by examination under anesthesia using fluorescein angiography. PMID- 8233391 TI - Randomized comparison of krypton versus argon scatter photocoagulation for diabetic disc neovascularization. The Krypton Argon Regression Neovascularization Study report number 1. AB - BACKGROUND: The Krypton Argon Regression of Neovascularization Study (KARNS) was designed to compare the efficacy of red krypton versus blue-green argon laser photocoagulation for the management of high-risk proliferative diabetic retinopathy. METHODS: A prospective, randomized clinical trial was performed in 24 clinical centers. Patients (n = 696, 907 eyes) with diabetes and neovascularization of the disc (NVD) of one-third disc area or greater in extent were assigned at random to either argon or krypton laser scatter (panretinal) photocoagulation. The major endpoint of the regression of NVD to less than one third disc area in extent at 3 months was evaluated by comparisons of gradings of the fundus photographs obtained at baseline and follow-up. RESULTS: At 3 months' follow-up, the proportion of eyes with regression of NVD to less than or equal to one-third disc area in extent were 41.1% in the argon-treated group and 41.8% in the krypton-treated group (P = 0.92). The odds of regressing to this extent of NVD at 3 months for argon-treated eyes versus krypton-treated eyes was 0.98 (95% confidence interval, 0.74-1.31). CONCLUSION: Scatter laser photocoagulation with either krypton red or argon appears to be equally effective in the treatment of proliferative diabetic retinopathy with NVD. PMID- 8233392 TI - Visual acuity in patients with best vitelliform macular dystrophy. AB - PURPOSE: Forty-seven patients with Best vitelliform macular dystrophy were evaluated in a cross-sectional fashion for visual acuity loss with age. METHODS: The authors assessed only patients who had at least one eye with a recognizable phenotype of Best vitelliform macular dystrophy. Patients with absent foveal changes or with only minimal foveal pigment mottling and hypopigmentation in each eye were excluded. RESULTS: A significant difference was noted between the visual acuities of the two eyes of the patients (2 lines or greater in the majority [64%] of patients). Nevertheless, for both eyes a significant correlation was noted between patient age and visual acuity, with older patients tending to have worse visual acuities. In the eyes with the best visual acuity, the majority of patients younger than 40 years of age (76%) had a visual acuity of 20/40 or better. In patients older than 30 years of age, a substantial percentage (74%) had a visual acuity of 20/100 or worse in at least one eye. CONCLUSION: The authors' findings indicate that although patients with Best vitelliform macular dystrophy who show characteristic macular lesions may retain good visual acuity in at least one eye, an appreciable number can lose substantial visual acuity, at least monocularly. In this population, no patient older than 50 years of age fulfilled the visual acuity criterion of 20/40 in at least one eye, the requirement in most states for an unrestricted driver's license, and only 20% of patients older than 40 years of age fulfilled this visual acuity criterion. PMID- 8233393 TI - Vitreous surgery for macular holes. AB - BACKGROUND: To surgically treat patients with macular holes, the authors previously reported both anatomic (re-attachment) and visual success (2 lines of improvement) in a series of 52 eyes. They now have operated on an additional 118 eyes using similar techniques, for a total of 170 eyes. METHODS: After ophthalmologic examination and history, the authors operated on suitable patients. The surgical objectives included relief of all tangential traction and retinal tamponade with intraocular gas. All eyes were followed for at least 6 months postoperatively. RESULTS: In the total population of 170 eyes, anatomic success was achieved in 73% and vision improved at least two lines in 55%. Twenty nine percent (49/170) of patients had a visual acuity of 20/40 or better at last examination. Patients with symptoms of less than 6 months' duration managed better than those with symptoms of longer duration (P = 0.3001). In the former group of 66 eyes, anatomic success was achieved in 80% (n = 53), whereas visual acuity improved at least two lines in 68% (n = 45) and at least four lines in 55% (n = 36). CONCLUSIONS: The authors suggest that macular hole surgery may provide meaningful improvement in visual acuity in most patients, especially in those whose symptoms are of less than 6 months' duration. PMID- 8233394 TI - Fine-needle aspiration biopsy of suspected intraocular tumors. The 1992 Urwick Lecture. AB - BACKGROUND: Fine-needle aspiration biopsy recently has been used as a diagnostic modality for selected intraocular tumors and simulating conditions. However, the value of fine-needle aspiration biopsy for intraocular tumors previously has not been clarified. METHODS: Transocular fine-needle aspiration biopsy was performed on selected patients who had intraocular lesions that were suspected clinically to be neoplasms but in which there was diagnostic uncertainty based on noninvasive clinical evaluation. These cases were analyzed to determine accuracy, complications, and limitations of the technique. RESULTS: Of 6500 patients referred to the Oncology Service for evaluation of possible intraocular tumor, transocular fine-needle aspiration biopsy was used clinically in 159 cases (2.4%). It proved to be a reliable diagnostic method for intraocular malignancies such as uveal melanoma, uveal metastasis, retinoblastoma, lymphoma, and leukemia. In the 140 cases (88%) in which adequate cytologic material was obtained, the sensitivity rate was 100% and the specificity rate was 98%. In 19 cases where the cytologic material was too scant to render a diagnosis, the sensitivity rate was 84% and the specificity rate was 98%. The problem of insufficient material for cytologic diagnosis has been greatly minimized with the recent use of a 22-gauge needle. The main complication was localized intraocular hemorrhage. Retinal detachment and tumor recurrence have not been observed. CONCLUSIONS: Transocular fine-needle aspiration biopsy is a safe and reliable diagnostic method for suspected intraocular tumors and inflammatory conditions in which noninvasive diagnostic modalities have failed to establish the diagnosis and in which cytologic verification of the diagnosis is necessary to institute appropriate treatment. Although the authors have had few complications with fine-needle aspiration biopsy, the technique should be reserved for selected cases where the diagnosis has not been established by less-invasive diagnostic measures. PMID- 8233395 TI - Long-term visual morbidity of cytomegalovirus retinitis in patients with acquired immune deficiency syndrome. AB - BACKGROUND: Patient survival with the acquired immune deficiency syndrome (AIDS) and cytomegalovirus (CMV) retinitis is increasing because of earlier diagnosis and improved medical therapy. Visual morbidity that occurs with prolonged survival has not been well described. METHODS: To evaluate the progression of retinitis, ocular complications, and visual morbidity, the authors retrospectively reviewed the records of 22 patients who had tested positive for human immunodeficiency virus since December 31, 1987. Each patient had an ophthalmologic diagnosis of CMV retinitis and had survived for a minimum of 6 months after diagnosis of retinitis. Patients were treated with intravenous ganciclovir alone, ganciclovir followed by foscarnet, or foscarnet alone (1 patient). RESULTS: Mean survival was 16.7 months after the diagnosis of retinitis. The retinitis progressed in 74% of eyes during therapy. Visual acuity of 20/70 or worse occurred in 79% of eyes at a mean of 7.6 months from diagnosis of CMV retinitis. No light perception occurred in 49% of eyes after a mean of 15 months patient survival. Eleven patients with visual acuity of 20/70 or worse in both eyes survived a mean of 11 months. In 39 eyes with CMV retinitis, the following complications occurred: retinal detachment (33%), papillitis (32%), branch retinal artery occlusion (10%), persistent iritis (5%), and cataract (2.5%). CONCLUSION: Improved modalities of therapy will continue to increase the survival of patients with AIDS and CMV retinitis. Progression of the retinitis occurs with current therapeutic regimens in the majority of patients. As survival increases, significant visual loss and ocular complications may compromise patient care and quality of life. PMID- 8233396 TI - Bilateral optic nerve cryptococcosis in sudden blindness in patients with acquired immune deficiency syndrome. AB - PURPOSE: A neuroanatomic study was undertaken to search for the cause of sudden, simultaneously bilateral blindness in a patient with acquired immune deficiency syndrome who had cryptococcal meningitis. METHODS: Careful gross examination was performed, and microscopic sections were cut at 50- to 100-microns intervals of the entire visual pathway. RESULTS: Focal cryptococcosis destroyed segments of the right intracanalicular optic nerve and the left intraorbital optic nerve adjacent to the optic canal. The meninges were heavily infiltrated by Cryptococcus organisms around the optic tracts, optic nerves, and optic chiasm; however, only a few scattered cryptococcal organisms were found in the periphery of the chiasm contiguous with heavy meningeal infection. Blood vessels supplying the chiasm appeared normal. Generalized cerebral edema and focal vacuolization of periventricular white matter were evident. CONCLUSION: The authors believe that sudden, simultaneously bilateral visual loss in this patient was caused by focal but fulminant necrosis of both optic nerves. However, the presence of cryptococcal organisms throughout the basal meninges and in the sheaths of both optic nerves suggests that cryptococcosis may produce visual loss by damaging multiple areas of the anterior visual pathway. PMID- 8233397 TI - Diffuse unilateral subacute neuroretinitis. Morphometric, serologic, and epidemiologic support for Baylisascaris as a causative agent. AB - PURPOSE: Several nematodes have been postulated as etiologic agents in diffuse unilateral subacute neuroretinitis (DUSN), but the cause of this condition remains uncertain. The authors report the first case of DUSN from the western United States (northern California), along with morphometric, serologic, and epidemiologic evidence supporting Baylisascaris procyonis as its cause. METHODS: One patient was examined and evaluated for disc edema and transient obscurations of vision. A diagnosis of DUSN was confirmed when a motile nematode was identified within the substance of the patient's retina. Morphometric analysis of the nematode was done from projected fundus photographs. Serologic evidence of Baylisascaris infection was suggested by Western blot analysis. A necropsy was done on 12 raccoons from the area. They were examined for evidence of Baylisascaris infection, previously believed to be nonendemic in the region. RESULTS: The intraocular nematode measured 1727 x 67 microns, most consistent with Baylisascaris. The patient had considerable exposure to raccoons, and was seropositive for B. procyonis infection on Western blot analysis. Necropsy evaluation showed B. procyonis infection in 8 of 12 raccoons examined from the area. CONCLUSIONS: The morphometric, serologic, and epidemiologic findings in this case provide evidence that the raccoon ascarid, B. procyonis, is a cause of the large nematode variant of DUSN. PMID- 8233398 TI - Orbital pseudotumor. Fine-needle aspiration biopsy and response to therapy. AB - PURPOSE: To determine the diagnostic accuracy of fine-needle aspiration biopsy and study the natural history and therapeutic response of a group of patients with orbital pseudotumor. METHODS: Retrospective review of patients seen by one ocular oncologist for diagnosis and management of presumed orbital pseudotumor. RESULTS: Thirty-eight patients had fine-needle aspiration biopsy under computed tomographic (CT) guidance. No false-positive results were observed. In 10% of fine-needle aspiration biopsies, negative results were observed. In all patients in this series, lymphomas could be differentiated from pseudotumors. Flow cytometry or Southern blot analysis could be performed on these cytopathologic specimens and was useful in selected cases. Eighty-one patients were followed after various forms of treatment. Essentially complete remissions were observed in 49 of 81 patients. High-dose oral corticosteroids were effective in approximately 31% of patients. A poor steroid response was observed in patients with mass lesions and those with fibrotic disease. Surgical resection was effective in 76% of patients; most of these had relatively discrete mass lesions. Radiation was effective in approximately 64% of patients. Radiation was more effective in nonfibrotic lesions and in patients with erythema. This type of treatment was less effective in fibrotic lesions and in patients who had a longer interval between diagnosis and therapy. CONCLUSIONS: Fine-needle aspiration biopsy is useful in evaluating patients with presumed orbital pseudotumor. There is a high success rate in many patients with orbital pseudotumor, although patients with diffuse fibrotic disease have a poor prognosis. PMID- 8233399 TI - Metastatic prostate cancer to the optic canal. AB - PURPOSE: Optic neuropathies represent a common cause of blindness among patients with cancer. The authors present the clinical and imaging findings and discuss the management of optic nerve involvement from metastatic prostate cancer to the optic canal. METHODS: Five patients with advanced metastatic prostate cancer had visual loss. Neuro-ophthalmologic examination and pre- and post-gadolinium orbital and brain magnetic resonance imaging (MRI) were performed. Compressive optic neuropathy secondary to extensive optic canal involvement was demonstrated in all of the patients. Neuropathologic examination was accomplished in two cases. A combination of steroid and focal radiation therapy was used for treatment in four patients. RESULTS: Imaging and neuropathologic findings demonstrated a compressive optic neuropathy secondary to both epidural metastases and to deformity and hypertrophy of the optic canal due to metastatic cancer. Treatment was of benefit in three cases. CONCLUSION: Acute and subacute unilateral or bilateral optic neuropathy in patients with prostate cancer may be the result of optic canal metastases. Magnetic resonance imaging shows a characteristic pattern of bone hypertrophy and deformity with optic nerve compression. Early combination steroid and radiation therapy may induce long lasting improvement. PMID- 8233400 TI - Apocrine adenocarcinoma arising in Moll gland cystadenoma. AB - CASE REPORT: An 85-year-old woman had a bluish tumor medially on the margin of the left upper eyelid. Malignant melanoma of the conjunctiva was presumed; the lesion was excised and submitted for routine histopathologic examination. There are no signs of recurrence or metastatic disease after follow-up for 1 year. METHODS: Light microscopy with routine and histochemical stains were used for diagnosis. Immunohistochemical analysis included the use of a broad-spectrum "cocktail" of cytokeratin antibodies (AE1/AE3). RESULTS: The features were those of a cystic cavity lined with apocrine epithelium in one part infiltrating the wall and consistent with an apocrine adenocarcinoma derived from a Moll gland cystadenoma. CONCLUSION: Malignant Moll gland tumors are extremely uncommon but may be confused with conjunctival malignant melanoma or metastatic disease. The differential diagnosis to other sweat gland tumors such as eccrine adenocarcinoma and benign apocrine tumor or to sebaceous carcinoma can be difficult. PMID- 8233401 TI - Primary basal cell carcinoma of the limbal conjunctiva. AB - BACKGROUND: The authors describe a 66-year-old man with primary basal cell carcinoma of the conjunctiva. He presented with a fleshy nodular conjunctival lesion of 4 months' duration, located at the nasal limbus of the left eye. No associated cutaneous lesions were present. RESULTS: The lesion was completely excised, and results of histopathologic examination showed a primary basal cell carcinoma of the conjunctiva. CONCLUSION: This report provides clinicopathologic evidence that primary basal cell carcinoma can rarely occur in the conjunctiva. The authors summarize two well-documented previously reported cases of this unusual lesion. PMID- 8233402 TI - Standardized color-coded maps for corneal topography. AB - PURPOSE: Modern videokeratoscopy is useful in assessing corneal shape. The purpose of this study is to compare color-coded topographic maps using standardized scales with 1.0- and 1.5-diopter (D) intervals. The authors assessed the use of the two scales for detecting clinically relevant features of corneal topography such as regular astigmatism, irregular astigmatism, early keratoconus, and contact lens-induced corneal warpage. METHODS: A total of 50 normal corneas, 50 corneas with contact lenses, 50 that had keratoconus (25 early to moderate and 25 advanced), 50 that had penetrating keratoplasty, 20 that had extracapsular cataract surgery, 17 that had excimer laser photorefractive keratectomy for myopia, 10 that had radial keratotomy, 3 that had aphakic epikeratophakia, and 2 that had myopic epikeratophakia were analyzed with a corneal topographic analysis system. Color-coded maps with 1.0-D intervals (Maguire/Waring scale) and 1.5-D intervals (Klyce/Wilson scale) were compared. RESULTS: There were no topographic characteristics that were not appreciated with either scale for corneas with dioptric powers that fell within their ranges. Conversely, for corneas that had powers outside the range of the 1.0-diopter scale, but within the range of the 1.5-diopter scale, the former produced a map in which the flattest or steepest areas were artifactually smoothed. CONCLUSIONS: This study suggests that the Klyce/Wilson scale (constant, 1.5-D intervals) provides the best combination of sensitivity for detection of clinically significant topographic features and the widest range of coverage of powers that are found on a variety of normal, pathologic, and surgically altered corneas. The adoption of the Klyce/Wilson scale by all manufacturers of corneal topographic instruments as a primary standard will facilitate communication and will make the interpretation of corneal topography easier for both the expert and the novice. PMID- 8233403 TI - Risk factors of corneal graft failure. AB - PURPOSE: To measure the association between potential risk factors and corneal graft failure. Two failure outcomes are compared: those with and those without a prior immune allograft reaction. METHODS: Based on a single-center observational study design, 539 adult recipients of a corneal graft were followed for a median time of 30 months. Survival analysis was carried out. RESULTS: Eighty-two graft failures were recorded. Of 82 failures, 53 (65%) were not preceded by an immune allograft reaction. Presence of blood vessels in the recipient cornea was associated with a twofold increase in risk for both failure outcomes. Three factors increased the risk of failure without an immune reaction: prior glaucoma or uveitis (adjusted relative risk estimate = 3.1), vitreous surgery with the graft (adjusted relative risk estimate = 2.0), and a repeat graft in the study eye (adjusted relative risk estimate = 2.0). Conversely, large graft wound size (adjusted relative risk estimate = 2.0). Conversely, large graft wound size (adjusted relative risk estimate = 2.9) and human leukocyte antigen (HLA)-A, -B incompatibility (adjusted relative risk estimate = 2.2) were associated with failures that followed an immune reaction. CONCLUSION: In this study, the authors support the clinical impression that corneal graft failures with and without a prior immune reaction are distinct phenomena. Enhanced surveillance in recipients with glaucoma and early intensive treatment of allograft reactions are recommended to improve the outcome of corneal grafts. PMID- 8233404 TI - Nd:YAG photodisruptors. American Academy of Ophthalmology. AB - The Nd:YAG laser surgery can cut lens capsule, vitreous and capsular membranes, strands, and adhesions, and the iris within the surgically unopened eye, thereby avoiding or minimizing infection, wound leaks, and other complications of conventional intraocular surgery. The technique has found its most widespread use in performing posterior capsulotomies after extracapsular cataract surgery. It has an extremely low complication rate when used in the anterior segment and is a preferred alternative to surgical discission. The uncertainties regarding its safety in creating iridotomies in phakic eyes have lessened with its extensive use in patients with pupillary-block glaucoma. However, caution is urged in other applications in phakic eyes. Following each Nd:YAG laser procedure, the eye should be monitored for elevation of intraocular pressure during the first two hours, and for retinal tears, retinal detachment, or cystoid macular edema during the first month after the procedure. Laser applications in the vitreous cavity appear to be of limited use. PMID- 8233405 TI - Considerations in the dental treatment of chemically dependent patients. PMID- 8233407 TI - Registered retirement savings plans (RRSPs rules and strategy for dentists). PMID- 8233406 TI - Management of the patient with a hyperactive gag reflex. Part one. PMID- 8233408 TI - Current concepts in the use of analgesics in dentistry. PMID- 8233409 TI - The higher cost of infection control. PMID- 8233410 TI - In search of excellence in endodontics. PMID- 8233411 TI - Registered retirement savings plans (more RRSP rules and strategies for dentists). PMID- 8233412 TI - Third molar surgery: current concepts and controversies. Part 1. AB - The removal of impacted third molar teeth ranks as the commonest procedure in the practise of oral and maxillofacial surgery. It is the purpose of this review article to present the major controversies and correct some of the misconceptions associated with third molar surgery. This paper deals primarily with the discussion of the indications for the removal while part II describes the influence of a variety of factors upon the outcome of third molar surgery. PMID- 8233413 TI - Third molar surgery: current concepts and controversies. Part 2. AB - The outcome of third molar surgery is influenced by a variety of factors including among others, mucoperiosteal flap design and flap closure and the use of antibiotics and corticosteroids. This article discusses the special significance of modifying factors upon the incidence of perioperative complications associated with third molar surgery. PMID- 8233414 TI - Aseptic practices in dental offices. PMID- 8233415 TI - The business side of dentistry/where is it going? PMID- 8233416 TI - Recurrent aphthous ulcers in association with HIV infection. PMID- 8233417 TI - Wide variation in patient response to midazolam sedation for outpatient oral surgery. AB - Intravenous midazolam in doses of between 0.07 mg/kg and 0.1 mg/kg has been recommended for sedation in dentistry and some medical procedures and investigations. This study examined the variable sensitivity of patients to midazolam. One hundred and thirty-four fit but anxious patients between the ages of 16 and 63 years received midazolam intravenously for sedation for minor oral surgery. Doses required ranged from 0.04 to 0.40 mg/kg. Nineteen patients required doses less than 0.07 mg/kg; 59 patients required doses greater than 0.1 mg/kg. The mean dose per kilogram required for males was significantly less than for females. The wide variation in sensitivity to midazolam is confirmed. PMID- 8233418 TI - Side effects and complications associated with third molar surgery. AB - A retrospective analysis of complications and side effects associated with surgery for 1000 mandibular and 500 maxillary impacted third molars was performed. The analysis included 614 patients with impacted mandibular third molars and 274 with impacted maxillary third molars who were treated in a 5-year timeframe from 1987 to 1992. The incidence of intraoperative complications and side effects of mandibular third molar surgery was 1.1% and 4% for maxillary third molar surgery whereas postoperative complications were 4.3% and 1.2%, respectively. PMID- 8233419 TI - Long-term progression of a traumatic bone cyst. A case report. AB - Through a series of similarly produced panoramic films, a chronologic picture of a traumatic bone cyst is demonstrated. This case represents a progressive rather than static lesion. The progression of the lesion is indicative of a etiologic process other than the solitary inciting event usually associated with traumatic bone cysts. PMID- 8233420 TI - Infrared laser soft tissue ablation versus ultraviolet excimer laser. Experimental introduction of the Hol:YAG-laser in oral surgery. AB - The in vivo tissue ablation characteristics of a pulsed infrared laser (Hol:YAG, lambda = 2120 nm) and a pulsed excimer laser (XeCl, lambda = 308 nm) were studied in an animal model. Laser energy was delivered via nylon fibers for the Hol:YAG laser and via quartz fibers for the excimer laser. Laser incisions were made under precise reproducible conditions on the sublingual side of the tongue and the gingiva of white rats. Laser surgery was done at two different energy output settings for the Hol:YAG laser and at one setting for the excimer laser. Histologic studies revealed tissue defects with clean contours for both laser types with small zones of necrosis of the adjacent tissue (Hol:YAG: 180 microns to 640 microns; excimer: 40 microns to 160 microns) and without carbonization. Both laser types function on the principal of photoablation and permit excellent control of tissue ablation. Wound healing was studied over a 10-day period and showed complete wound closure by re-epithelialization. The in vivo tissue ablation characteristics and the surgical reliability of the two lasers are compared and discussed with respect to oral and periodontal surgery. PMID- 8233421 TI - Benign osteoblastoma associated with an aneurysmal bone cyst of the mandibular ramus and condyle. AB - This article reviews the clinical behavior, the histologic conditions, and the treatment of a benign osteoblastoma associated with an aneurysmal bone cyst of the mandibular condyle and ramus. A 14-year-old boy, otherwise healthy, was referred for pain and enlargement in his right temporomandibular joint-cheek region. The swelling was firm and tender, a slight facial asymmetry was present, and the mouth-opening capacity was reduced. During a 6-weeks period from the primary examination to surgery the lesion expanded extensively, anesthesia appeared in the right inferior alveolar nerve and a lateral open bite developed on the affected side. Computed tomography displayed a total destruction of the right mandibular condyle and ramus to the level of the mandibular foramen. The lesion was radiolucent without distinct borders to adjacent bone. Surgery revealed a bluish, well-vascularized predominantly cystic tumor. The lesion included the temporomandibular joint disk but without overgrowth to the temporal component. After resection of the lesion, the condyle and the mandibular ramus was reconstructed with the use of an autogenous costochondral graft. The postoperative healing was uncomplicated. At follow-up 2 years after surgery the boy was free of recurrence with normal motor and sensory nerve function. PMID- 8233422 TI - Lesch-Nyhan syndrome and the lower lip guard. AB - Lesch-Nyhan syndrome, a rare inborn error of metabolism, is characterized by mental retardation and factitious oral lesions, which cause immense difficulties in its management. A case of an 11-year-old boy, the elder of two siblings with this condition, who had severe self-inflicted injuries to oral and perioral tissues is presented highlighting the different treatment modalities considered. A tailor-made lipguard proved to provide the best cosmetic result short term. PMID- 8233423 TI - Effects of chlorhexidine during immunosuppressive chemotherapy. A preliminary report. AB - Immunosuppressive chemotherapy and bone marrow transplantation are increasingly used to treat and, in some cases, cure numerous malignant conditions. The systemic sequelae as a result of these immunosuppressive techniques lead to a host of oral and dental complications. The direct and indirect stomatotoxic effects lead to development of ulcerative, hemorrhagic, or infectious complications that potentially can lead to increased mortality and morbidity. Chlorhexidine was studied to evaluate its broad spectrum antimicrobial properties and to evaluate its effect on immunosuppression-induced mucositis. Our double blind randomized study revealed that chlorhexidine versus controls had considerable effect in the bone marrow transplant cohort. Patients who received chlorhexidine while undergoing bone marrow transplantation were found to exhibit fewer and less painful mucositis lesions. This study supports prophylactic use of chlorhexidine in patients who undergo bone marrow transplantation. PMID- 8233424 TI - Audit of diagnosis and investigations in patients with recurrent aphthous stomatitis. AB - The responses of clinicians to results of hematologic and serologic investigation of 79 patients with recurrent aphthous stomatitis have been reviewed. Twenty percent of patients with abnormal results were not adequately managed by attending clinicians. PMID- 8233425 TI - Clinical, historic, and therapeutic features of cicatricial pemphigoid. A literature review and open therapeutic trial with corticosteroids. AB - Cicatricial pemphigoid is at present an incurable, autoimmune disease that involves mucosa and skin. We have documented the clinical, microscopic, and therapeutic features of 23 patients with cicatricial pemphigoid and added these to reports of past literature. The mean age at the time of diagnosis was 63 years, and women were involved twice as often as men. Eighty-three percent of patients had oral mucosal involvement, 70% had conjunctival involvement, and 22% had skin involved. Direct immunologic evaluation revealed IgG at the basement membrane in 57% of cases and C3 in 66%. We have been able to manage the disease adequately in most patients with topical or systemic corticosteroids. The most commonly encountered side effect was oral candidiasis. PMID- 8233426 TI - White lichenoid lesions of the buccal mucosa in patients with HIV infection. AB - We report on eight patients who developed white lichenoid lesions of the buccal mucosa during the course of human immunodeficiency virus infection. In five patients the lesions appeared after the administration of zidovudine, in two after the intake of both zidovudine and ketoconazole, and in one after ketoconazole. In the majority of cases, lesions presented as bilateral reticular keratosis or atrophic changes of the buccal mucosa. Three patients manifested lichenoid atrophic changes of the dorsum of the tongue. The histopathologic features were hyperkeratosis, epithelial atrophy, basal cell liquefaction, and the presence in the lamina propria of either a patchy or diffuse lymphocytic infiltrate. All specimens tested negative for Epstein-Barr virus and human papillomavirus. Our study suggests that lichenoid lesions of the buccal mucosa, similar to what has been described as lichenoid drug reactions or idiopathic lichen planus, can be observed during human immunodeficiency virus infection and that administration of zidovudine and ketoconazole should be considered as a possible cause. PMID- 8233427 TI - Morphology of the bone that supports endosteal dental implants. Transmission electron microscopic and high voltage electron microscopic observations. AB - The morphologic features of the bone-dental implant interface were investigated using an in vivo dog model. The undecalcified bone and associated support tissues were serially sectioned and examined with both conventional and high voltage transmission electron microscopy. A varied morphologic appearance of the tissues supporting clinically and radiographically appearing integrated implants was observed. Osteoblasts were observed at the implant interface, and osteocytes were routinely seen encased within lacunae extremely close to the implant surface. Often these osteocytes extended cellular projections to the implant surface. The variable tissue types observed were suggestive of healthy lamellar and appositional type mineralization patterns adjacent to the implants. PMID- 8233428 TI - Histocompatibility antigens and geographic tongue. AB - The HLA-A, B and HLA-DR antigens were investigated in 50 unrelated Greek persons with geographic tongue and in 380 healthy control persons. An increased incidence of DR5 and DRW6 antigens was observed in the blood of persons with geographic tongue. Ten (20%) of them had the DRW6 antigen, and it was significantly increased compared with 29 (7.6%) of the controls (p < 0.01, RR = 3.32). Twenty seven (54%) of the experimental group showed DR5 antigen compared with 136 (35.7%) of the controls (p < 0.025, RR = 2.18). On the contrary, only 12% (5) of the experimental group had the B51 antigen in comparison with the controls (26.3%) (p < 0.05, RR = 0.37). A decrease of the DR2 antigen was also found in the persons with geographic tongue (24%) in comparison with the controls (39.2%) (p < 0.05, RR = 0.58). PMID- 8233429 TI - Clinical diagnosis of caliber-persistent labial artery of the lower lip. AB - Only 14 cases of caliber-persistent labial artery of the lower lip have been reported to date. Six of these were misdiagnosed and treated as squamous cell carcinoma, another as a mucocele. The correct diagnosis emerged only after the wedge resections were examined histopathologically. We report the first cases of caliber-persistent labial artery to be diagnosed clinically since the original description of the condition by Howell and Freeman in 1973. Our first case was a nonpulsatile hard, linear, "gooseneck lamp" submucosal nodule of the lower lip. On the basis of an initial misdiagnosis of sclerosing sialadenitis, a biopsy was attempted. Brisk pulsatile bleeding proved the lesion to be an artery, and the superficial location and large diameter of the vessel lead to the clinical diagnosis of caliber-persistent labial artery. The "gooseneck lamp" hardening is typical of Monckeberg's arteriosclerosis. The second case was a pulsatile blue linear submucosal nodule of the lower lip. The clinical diagnosis of caliber persistent labial artery was confirmed when angiography showed the lesion to be an abnormally dilated labial artery. Both cases were successfully ligated with no complications at 16 and 10 months after surgery. PMID- 8233430 TI - Human pulpal response to hydroxyapatite and a calcium hydroxide material as direct capping agents. AB - Forty maxillary and four mandibular first premolars that were to be extracted for orthodontic reasons were used in this study to evaluate human pulpal response to hydroxyapatite and Dycal as direct capping materials. After the postoperative periods of 2 days, 30 days and, 60 days, teeth were extracted and prepared for the histologic examination. The results of this study showed that hydroxyapatite does not induce hard tissue bridging at the exposure site in human dental pulp. No hard tissue bridging at the exposure site was observed in any of the teeth capped with hydroxyapatite. Moderate infiltration of the inflammatory cells was seen generally in the teeth capped with hydroxyapatite. However, this study has verified that exposed human dental pulp will heal under Dycal application with hard tissue bridging and with none or minimal inflammatory response. PMID- 8233431 TI - Tissue reactions to endodontic materials implanted in the mandibles of guinea pigs. AB - Teflon, Intermediate Restorative Material, and Prisma VLC Dycal were implanted in the mandibles of 16 guinea pigs. After experimental periods of 4 and 12 weeks, the animals were killed and tissue sections were prepared. No severe inflammatory response was noted in any of the groups except for a persistence of moderate inflammation in the poorly condensed Teflon group. Bone apposition occurred in the Prisma VLC Dycal group; none-to-mild inflammation and a thin fibrous capsule characterized the IRM group; and the Teflon group had a thick fibrous encapsulation except for samples where the Teflon material appeared to be poorly condensed and dispersed into the bony tissue. In areas where the teflon was loosely condensed, chronic inflammation and active phagocytosis were seen along the surface of the material. PMID- 8233432 TI - 35-mm film scanner as an intraoral dental radiograph digitizer. I: A quantitative evaluation. AB - A 35-mm slide scanner digital imaging system was tested for its suitability in digitizing intraoral dental radiographic film for quantitative studies. The system (Nikon model LS-3510AF Nikon Electronic Imaging, Nikon, Inc., Melville, N.Y.) uses a charge-coupled device linear photodiode array. The data content in the original film images was evaluated, and the system performance assessed objectively with the use of specially designed test films. Radiometric and geometric performances for the digitizing system were extracted from measurements and observations, and these were compared with published data for two other film digitizing systems (video camera DAGE MTI, Michigan City, Ind. and Barneyscan 35 mm film digitizer Barneyscan, Berkeley, Calif.). The techniques used to evaluate this system are easy and suitable for evaluation of any digitizing system. This scanner system (Nikon) was superior to previously evaluated systems in transforming and recording radiographic film densities across the range (0.3 to 2.0 optical density units) of clinically relevant optical densities. The scanner offers substantial advantage over the other digitizing systems for gray scale information from clinically important optical densities. PMID- 8233433 TI - 35-mm film scanner as an intraoral dental radiograph digitizer. II: Effects of brightness and contrast adjustments. AB - Typical 35-mm slide scanners use a photodiode array and software that allows for digital and analog controls that are manually adjustable. The digital controls provide brightness and contrast adjustments, whereas corresponding analog controls adjust the exposure time and black level that determines the clamping level of the charge-coupled device for the maximum black in the image. The objective of this study was to determine the effects of these controls on the radiometric data of intraoral dental radiograph images, to establish recommended settings, and to set specific standard guidelines for the digitization process. Three approaches were used. The results of this study demonstrate that brightness and contrast control alterations on the digitizer produces different optical densities and modulation transfer function values. The impact of these results is unresolved yet must be considered in analysis on quantitative radiometric studies. PMID- 8233434 TI - Absorbed doses with intraoral radiography. Function of various technical parameters. AB - Both the maxillary incisor and mandibular molar regions of a Rando phantom were radiographed using specific technical parameters. The parotid gland and thyroid gland doses were measured using thermoluminescence dosimetry. Tube voltage, total filtration, mAs, collimation, and cone length were varied while keeping the other factors constant. Increased tube voltage or total filtration generally resulted in a slightly increased absorbed dose. Absorbed dose was decreased as the beam was more tightly collimated or the cone length was increased. PMID- 8233435 TI - Radiologic features of desmoplastic variant of ameloblastoma. AB - Desmoplastic variant of ameloblastoma is a newly described variant characterized by unique histologic and radiographic features. This study reviews and analyzes the clinical and radiographic features of 15 cases of desmoplastic variant of ameloblastoma (14 cases from the literature and one case from our files). Desmoplastic variant of ameloblastoma shows a predilection for the maxilla and has a tendency to occur in the anterior region of the mandible and maxilla. Radiographically, almost all desmoplastic variant of ameloblastomas exhibit a mixed radiolucent-radiopaque appearance. Most show poorly defined or diffused borders and are mistakenly diagnosed as fibro-osseous lesions. PMID- 8233436 TI - Antral exostosis in panoramic radiographs. PMID- 8233437 TI - Clinical significance of the panoramic innominate line in the diagnosis of a maxillary sinus carcinoma. PMID- 8233438 TI - Focal osteoporotic bone marrow defect in the anterior maxilla. AB - An unusual presentation of a focal osteoporotic bone marrow defect is documented. A definitive diagnosis could not be established on the basis of the location, history, clinical, and radiographic appearance. A biopsy provided the identity of the lesion. PMID- 8233439 TI - Drug-induced gingival overgrowth. AB - Certain anticonvulsants, cyclosporine, and a variety of calcium channel blockers have been shown to produce clinically and histologically similar gingival enlargements in certain susceptible patients. These drugs appear to be similar with respect to their pharmacologic mechanism of action at the cellular level. The primary target tissue is the most essential difference among them. Therefore it is tempting to speculate that these agents may act similarly on a common secondary target tissue, such as gingival connective tissue, and cause a hyperplastic response. This tissue reaction may involve a disturbance of calcium ion influx into specific cell populations with a resulting alteration in collagen metabolism and other host cell response mechanisms. A connection between ion exchange, folate uptake, collagenase activation, and bacterial inflammation may exist. Until a more effective approach can be developed from future research results, treatment should continue to emphasize plaque control, professional debridement, and resective gingival procedures to improve function, esthetics, and access for home care. PMID- 8233440 TI - Efficacy of digital subtraction angiography for confirmation of position of the infusion tube for delivery of intra-arterial chemotherapy as treatment of the head and neck cancer. AB - This study was conducted in 47 patients with biopsy-proven cancer of the head or neck. These patients each had an intra-arterial infusion tube inserted retrogradely. Positioning in the infusion tubes had been determined by staining of the mucocutaneous area of the malignant lesion under observation with injection of blue dye. Digital subtraction angiography (DSA) was performed from the infusion tube. DSA demonstrated that infusion tubes were in the correct position in 26 patients (55%), and in an incorrect position in 21 patients (45%). The infusion tubes of all patients with cancer of the maxillary sinus were in the correct position. However, the majority (54%) of infusion tubes in patients with cancer of the tongue were in an incorrect position. Observation of arterial flow from the infusion tube using DSA should be made in all patients with head or neck cancer in order to avoid the potential complications of intra-arterial chemotherapy. PMID- 8233441 TI - Neuroanatomical analysis of peptidergic input in the spinal nucleus of bulbocavernosus. AB - The spinal nucleus of bulbocavernosus (SNB) is located in the anterior horn of the lower lumbar and upper sacral segments of the spinal cord and plays a significant role in the regulation of external sphincters and other muscles of the urogenital tract and rectum. The present study examined characteristic features of peptidergic input to this nucleus in rat using a double immunostaining method at both light and electron microscopic levels, focused on an analysis of relationship between cholinergic motoneurons and neuropeptide Y (NPY) axon terminals in the SNB. At the light microscopic level, NPY immunoreactive terminals were frequently distributed in the SNB and some were in the vicinity of the cholinergic cells. At the electron microscopic level, some of NPY immunoreactive terminals were seen to form synaptic contacts with cholinergic neurons on their somata and dendrites in the SNB. These findings suggest that NPY terminals are involved in the functioning of cholinergic SNB motoneurons via the synapse. PMID- 8233442 TI - Intravenous digital subtraction angiography of the internal jugular veins. AB - 92 patients with various head-related symptoms were underwent study with IV-DSA. The width and density of the internal jugular veins were compared with the width and density of the common carotid arteries. The right internal jugular vein was more than twice as broad as the common carotid artery in 58 patients (63%), and was denser than the common carotid artery in 14 patients (15%). On the other hand, the left internal jugular vein was broader than the common carotid artery in 76 patients (83%) and more than twice as broad as the common carotid artery in 19 patients (21%). However, the left jugular vein was narrower than the common carotid artery in 12 patients (13%), and in 5 patients (5%) could not be demonstrated. If it is recalled that the internal jugular veins are sometimes congenitally aplastic or hypoplastic as developmental variations of the confluence of sinuses, the occurrence of complications associated with retrograde catheterization can be minimized. PMID- 8233443 TI - [Insulin-like growth factors]. PMID- 8233444 TI - [The place of insulin-like growth factor I in the diagnosis of acromegaly]. AB - Insulin-like growth factor-I (IGF-I) and basal growth hormone (GH) (average of GH values measured at -20 an 0 minutes) levels were measured in 32 (29 treated, 3 untreated) patients with acromegaly. In 15 patients IGF-I and basal GH values were normal. In 12 patients both GH and IGF-I were elevated. In 5 cases normal basal GH and elevated IGF-I levels were found. We assume that elevated serum level of IGF-I is a reliable sign of acromegalic activity. Compared to this, the basal GH level proved to be a much less reliable indicator to predict or exclude active acromegaly (sensitivity: 0.7, specificity: 1, positive predictive value: 1, negative predictive value: 0.75). PMID- 8233445 TI - [Which patients are dying of tuberculosis in the 90-s?]. AB - Medical and social characteristics of 27 patients deceased in tuberculosis in the Koranyi National Institute for Tuberculosis and Pulmonology, Budapest, were analysed and evaluated. M. tuberculosis strains sensitive to all antituberculotic drugs were isolated from all cases. Characteristics of patients: males over 40 years of age (17 cases), single (9 cases), alcoholics (16 cases) who were transferred from another hospital (13 cases). All patients had pulmonary tuberculosis, in 3 cases together with extrapulmonary lesions. Sputum direct smear examination revealed acid-fast bacteria in 9 cases. There were 8 relapses. It is proposed to consider possibility of tuberculosis in the presence of the above mentioned "risk factors". PMID- 8233446 TI - [The development of spinal movements and sagittal curves in postural defects, idiopathic scoliosis and Scheuermann disease (results of longitudinal studies on 5, 10 and 14 year old subjects)]. AB - A given population was followed by authors for 10 years. Children suffering from postural defects, structural scoliosis, spinal osteochondrosis were separated from a normal group. Spinal mobility and posture were compared with the mobility of normal, healthy boys and girls aged 5-14 years. In postural defects physiologically curves could be seen. The range of motion hasn't been physiologic in structural scoliosis. The rotation and flexion of the spine were increased while the lumbar flexion was decreased. Physiologic trend was found in Scheuermann disease, however high values of spinal rotation wasn't found. Extremely increased thoracic kyphosis was found only in children aged 14 years. Perhaps the change of motion is an etiologic factor in structural scoliosis. PMID- 8233447 TI - [A case of fatal squamous cell carcinoma, originating from the skin of the hand]. AB - Case report of a 77 year old man has been shown, who had a squamous cell carcinoma of the left hand, with axillary metastases. Seven months after the recognition of disease, incision of primary tumor, skin transplantation and removal of axillary tumor mass, the patient has died. PMID- 8233448 TI - [53 years of activity of a Hungarian physician in Spain (Fulop Hauser, 1832 1925)]. PMID- 8233449 TI - [Pal Bugat, reformer of Hungarian medical terminology]. PMID- 8233450 TI - [Heart diseases and blood groups]. PMID- 8233451 TI - [Neonatal neuroblastoma revealed by ultrasonic screening]. PMID- 8233452 TI - [Physicians in the Vac prison hospital 1958-63]. PMID- 8233453 TI - [Screening of congenital anomalies in the mid-trimester of pregnancy. Prospective epidemiologic study. Developmental anomalies]. AB - Authors report about the data of 60,000 pregnant women from three district counties of Eastern-Hungary between 1988 and 1990, on whom both ultrasound and alfa-fetoprotein screenings were performed. They demonstrate the value of this screening for the detection of fetal malformations and for the birth prevalence rate as well. This effective screening-protocol is proposed for a nation-wide application. PMID- 8233454 TI - [Spiro-ergometry in chronic obstructive lung diseases]. AB - The relationship of ventilatory function (FEV1, RV) with ergospirometric parameters (VO2max, VEmax/MVV) and arterial blood gas changes (delta paO2, delta paCO2) was investigated in 52 COPD patients. After exercise group I patients (n = 32) had better oxygenation (paO2: 62.6 +/- 7.2-->78.2 +/- 11.2 mmHg, P < 0.001), while paO2 in group II (n = 20) deteriorated (60.9 +/- 7.3-->54.1 +/- 5.4 mmHg, p < 0.001). Significant correlation have been found only in group II between FEV1 and VO2max (r = 0.65, p < 0.01) and RV and VO2max (r = -0.64, p < 0.01). Of the 41 patients who reached anaerobic threshold (AT), only 8 showed worse paO2 after exercise. Our results suggest that simultaneous observation of metabolic (AT) and respiratory (paO2) parameters during exercise are necessary to select those COPD patients who could have benefit from high intensity regular physical training. Those, who are able to reach AT and paO2 does not deteriorate during exercise are the best candidates for this kind of treatment. PMID- 8233455 TI - [Experience with Tenckhoff catheter implantation in chronic peritoneal dialysis]. AB - The authors performed 310 Tenckhoff catheter implantations during the last 17 years. They had 64 complications, and 29 cases the catheter had to be changed. The incidence of peritonitis was 0.22 patients/year. They present the technic of implantation, the complications, and the possible abdominal surgeries with the patients under dialysis treatment. PMID- 8233456 TI - [Diabetes mellitus and lipoproteins]. AB - The leading cause of death of diabetic patients is coronary heart disease developing on the basis of accelerated arteriosclerosis. Lipoprotein disorders commonly accompanying diabetes mellitus (DM) promote this process and their joint presence represent cumulated risks for patients. The present review summarizes the various disorders accompanying the two types of DM. In case of insulin treated, type I, well adjusted DM without signs of primary disturbances of lipid metabolism, quantitative lipid alterations cannot be found. In poorly treated DM with insufficient insulin levels the accumulation of triglyceride particles is due to diminished lipoprotein lipase activity. The main consequence is hyperchylomicronaemia but all three lipoprotein classes are affected. The most characteristic lipid abnormalities in non-insulin-dependent (type II) DM are hypertriglyceridaemia, increased VLDL cholesterol concentration and decreased HDL levels, which frequently remain unchanged even upon the proper treatment of glucose metabolism. The alterations are related to increased free fatty acid levels and decreased glucose uptake resulting from elevated insulin levels. In about one-fourth of cases, primary hyperlipaemia is also present. The treatment of primary and secondary lipid disorders accompanying DM by diet and drugs is of the most uttermost importance. PMID- 8233457 TI - [Laryngopharyngectomy and free jejunal graft reconstruction in the treatment of hypopharyngeal tumors]. AB - The authors performed a total laryngopharyngectomy due to a hypopharyngeal neoplasm. The continuity of the alimentary tract was restored by the insertion of a vascularized free jejunal graft, using microsurgical technique. On the occasion of this case the authors give a survey of the therapeutical methods used in the therapy of hypopharyngeal tumors, of the indications of the free jejunal transplant and of the technical issues and complications of this procedure. PMID- 8233458 TI - [Informal conversation with colleagues about drugs, prices, as well as common duties and obligations]. PMID- 8233459 TI - [Unusual ultrasonic image of the gallbladder]. AB - It often occurs during ultrasonography, that the gallbladder is not visible. The cause of this is most frequently cholelithiasis, but one must know the less common causes as well. Authors review the characteristic reasons and give accounts of their own experience, which was gained in the course of more than 60,000 examinations. PMID- 8233460 TI - [Certain aspects of continuous ventilation]. AB - The indication of mechanical ventilation is either a failing ventilatory function of the thorax or a failing gas exchange function of the lung. The ventilation affects every organ function in more or less degree. The starting point of long term ventilation is debated, arbitrary. Some ventilatory devices are simulating the natural intrapleural negative pressure principle, but most of them applies intermittent positive pressure into the lung. For improving oxygenation, a moderate level of positive end expiratory pressure, eventually inversed or 1:1 inspiratory:expiratory ratio can be applied. The computerized electronic ventilators offer a big selection of assisting and controlling ventilatory modes, according to the requirements of the patients. For routine use, in ventilatory failure, pressure cycled controlled ventilation, in gas exchange failure, the combination of synchronized intermittent mandatory ventilation, positive end expiratory pressure and pressure support can be recommended. For special tasks there are existing special ventilatory or other supportive means. In most of the cases some sedation of the ventilated patients is appropriate, muscle paralysing is restricted to a few situations. It is important to humidify the inhaled air. The detailed monitoring of the patients is essential during ventilation. The weaning of the patients from the ventilator is a complex procedure, which involves ventilatory, nutritional, pharmacologic and psychologic interventions. PMID- 8233461 TI - [Fetal chromosome abnormalities diagnosed by chorionic villi sampling]. AB - Chorionic villus sampling was performed for chromosome analysis in 387 cases during a 4-year-period. In 115 cases transcervical while in 272 cases transabdominal sampling was carried out. Chromosomal abnormalities were found in 25 cases (6.4%). Autosomal trisomies occurred in 17 cases, structural anomalies in 2 cases and sex chromosomal aberrations in 6 cases. The pregnancy was terminated in 19 cases because of chromosome abnormality, in 5 further cases because of X-linked disease and male fetus. After transcervical sampling spontaneous abortion occurred in 7 cases (5.8%), while after transabdominal sampling in 8 cases (2.8%). The authors prefer in their practice the early transabdominal CVS, which can be performed safety already at the end of the first trimester. PMID- 8233462 TI - [Giant coronary thrombus as a complication of Kawasaki disease]. AB - We report the case of a 3 yrs old boy with the most serious complication of Kawasaki disease. 2 dimensional echocardiography detected multiple coronary aneurysms and in the giant left coronary aneurysm a thrombus formation. The thrombus disappeared after one week anticoagulant therapy. In addition the aneurysm were proved by coronarography. During the 2 years follow-up period the child is on anticoagulant therapy and he is symptom free. This is the first case in our country with a huge thrombus in a giant coronary aneurysm due to Kawasaki disease who was successfully treated using anticoagulant therapy. PMID- 8233463 TI - [Development of Hungarian medical publications]. PMID- 8233464 TI - [Psychology and psychopathology of love in literature and forensic psychiatry]. PMID- 8233465 TI - [Drug therapy of iatrogenic hypothyroidism in patients with differentiated thyroid cancer]. PMID- 8233466 TI - [The right of the patient]. PMID- 8233467 TI - [The right of the patient]. PMID- 8233468 TI - [Management of pathological fractures in bone metastases. Principles, methods, results]. AB - The data of 96 patients treated for impending or existed fractures between 1980 1990 are discussed in this paper. The authors describe a practical method of treatment based on a simple staging. The analysis of the survival times support the applicability of this recommended method. The authors suggest to the importance of the early diagnosis and draw attention to the control with scintigraphy of the tumor patients. The necessity of the surgical treatment of the patients suffered from pathological fractures is nowadays hardly debatable. The early diagnosis and the "radical" surgery improve the patient's chances for survival and can better the quality of life as well. PMID- 8233469 TI - [Complex therapy of cystine calculi]. AB - Authors report their experiences with management of cystine urolithiasis between 1975-1992. 61 patients were observed and treated with dietotherapy, medicaments, open surgery, percutaneous nephrolithotomy and extracorporeal shock wave lithotripsy. Difficulties of treatment of this rare stone have been demonstrated with data of 3 patients (lack of screening and early diagnosis, unsystematic prophylaxis, formation of large stones). PMID- 8233470 TI - [Needle biopsy of the pleura]. AB - A total of 171 pleural needle biopsies were carried out in 123 patients with pleural effusion. The sensitivity of the method proved to be 51% for malignant diseases and 75% for tuberculotic pleurisy, respectively. Performing biopsy simultaneously with the cytological examination of the pleural fluid, malignancy was proved only by needle biopsy in 20% of the patients, while cytology was found negative in these cases. Simultaneously performed thoracocentesis and pleural needle biopsy is suggested. PMID- 8233471 TI - [Clinical significance of lumbosacral transitional vertebrae (Bertolotti syndrome)]. AB - 500 lumbosacral plain x-ray injured patients and 1000 myelography of patients suffering from low back pain and nerve root sign were reviewed to evaluate the rate of transitional lumbosacral vertebra. This anomaly appeared in 4.6% in the normal population. The rate of the transitional vertebra in patients with nerve root syndrome was double (8.4%) whereas that with lumbar disc herniation four time as many as in the normal population. The data demonstrate that the asymmetrical anomalies have more clinical significance. The authors consider, that the role of the transitional vertebra is twofold: it can be consider primary in low back and sciatic pain, but in the formation of lumbar disc herniation is secondary. They find the analysis of the lumbosacral plain x-ray is necessary even in cases with CT and MRI. PMID- 8233472 TI - [Pericardial-peritoneal shunt]. AB - A new shunt procedure is reported for persistent pericardial effusions. Abdominal approach to pericardial sac via transphrenic window is described. The technique proved to be useful in cases when the standard pericardiopleural procedure was not the proper solution. A decision-making algorithm is suggested when one faces different types of pericardial tamponade. PMID- 8233473 TI - [Sandor Koranyi in Hungarian medicine]. PMID- 8233474 TI - [Centenary of the Debrecen Municipal Hospital. Message to the participants of the bicentenary celebration]. PMID- 8233475 TI - [Position of the Work Group for Nursing in the Operating Room Service]. PMID- 8233476 TI - [Rosa Ecker-Zohrer: "publicity work for even more initiative. Interview by Harald Verworner]. PMID- 8233477 TI - [Sexual relations in nursing]. PMID- 8233479 TI - [Sexual activity in old age]. PMID- 8233478 TI - [Dialog with Mrs. Margarethe Lindner, psychotherapist, psychoanalyst. Interview by Doris Hoffmann and Harald Verworner]. PMID- 8233480 TI - [Sexuality and the image of the nursing personnel: Madonna, witch, prostitute, gay, impotent]. PMID- 8233481 TI - [Sexuality in nursing]. PMID- 8233482 TI - [They all were laughing uproariously]. PMID- 8233483 TI - To be or not to be a risk-taker. PMID- 8233484 TI - Aminoglycoside cochlear ototoxicity. AB - The aminoglycoside antibiotics streptomycin, kanamycin, neomycin, gentamicin, tobramycin, amikacin, and netilmicin are discussed in this article on cochlear toxicity of aminoglycosides. Topics discussed include pharmacokinetics, comparative studies on toxicity, toxicity in neonates and children, histopathology, and aminoglycoside monitoring. PMID- 8233485 TI - Vestibular ototoxicity. Clinical considerations. AB - Ototoxicity may arise from a variety of medications, chemical solvents, or environmental compounds. The resulting changes in auditory or vestibular function may cause significant morbidity as well as devastating effects on socioeconomic productivity and overall quality of life. Through careful attention to drug selection, dose, monitoring, and antecedent risk factors, ototoxic damage can be minimized or even prevented. Otolaryngologists play a critical role in identifying ototoxicity and in the treatment and rehabilitation of affected patients. PMID- 8233486 TI - Therapeutic use of aminoglycosides in Meniere's disease. AB - This article details the various effects of aminoglycosides used to treat individuals with Meniere's disease. Effects of aminoglycosides on hair cells and dark cells are described in addition to intramuscular streptomycin, intratympanic application of aminoglycosides, and the application of streptomycin to the lateral semicircular canal. PMID- 8233487 TI - Ototoxicity of topical agents. AB - Topical otic preparations are commonly prescribed by otolaryngologists--head and neck surgeons. It is well documented in the literature that some agents in these preparations are ototoxic when applied to the middle ear in experimental animals, although the occurrence of ototoxicity in the clinical setting is controversial. This article discusses topical otic preparations currently available as well as various other agents. The clinician should prescribe these preparations with close surveillance for potential ototoxicity. PMID- 8233488 TI - Ototoxicity of chemotherapeutic agents. AB - This chapter summarizes the reported ototoxicity data on the most clinically important ototoxic chemotherapeutic agents, notably emphasizing the oto(neuro)toxicities of the more commonly administered platinum compounds, cisplatin and carboplatin. Currently, in the United States, the only other marketed ototoxic chemotherapeutic agents are nitrogen mustard, alpha difluoromethyl ornithine (DFMO), and the vinca alkaloids (vincristine and vinblastine sulfate); for these groups, animal ototoxicity data is sparse, and audiovestibular records of human ototoxicity are not available from most prospective, randomized controlled clinical trials. Future phase I, II, and III clinical oncologic trials of "experimental" chemotherapeutic agents should include methodology for audiovestibular monitoring, just as present FDA-approved cancer protocols with either monotherapy or combined therapy of known "ototoxic" agents should include standardized audiovestibular assessment in the database. Finally, continued clinical application of cisplatin alone or in combination with other chemotherapeutic agents in the successful treatment of solid tumors mandates decreasing or eliminating specifically the dose-dependent sensorineural hearing loss (partially in cases of complete or long-term partial remission) in addition to other common antiproliferation-induced side effects (nephritis, peripheral neuropathy, intractable nausea and vomiting, electrolyte imbalance, anaphylactic-like reactions, and myelosuppression). Because of the chemotherapeutic superiority of cisplatin, it is essential to continue to investigate methods of altering the dose-limiting oto(neuro)toxicity without causing a "counterproductive" reduction of the antitumor activity of cisplatin (or other second- or third-generation ototoxic platinum agents). PMID- 8233489 TI - Ototoxicity of salicylate, nonsteroidal antiinflammatory drugs, and quinine. AB - Salicylates and most NSAIDS in high doses cause mild to moderate temporary hearing loss, either flat or greater in the high frequencies. Hearing loss is accompanied by tinnitus and suprathreshold changes. Salicylates may or may not exacerbate hearing loss and cochlear damage induced by noise. The mechanism of salicylate ototoxicity seems to be multifactorial. Morphologic studies suggest that no permanent cochlear damage occurs with salicylate ototoxicity. Electrophysiologic, morphologic, and in vitro data conclusively demonstrate that salicylate affects outer hair cells. In addition, salicylates appear to decrease cochlear blood flow. Salicylates and NSAIDs inhibit PG-forming cyclooxygenase, and recent studies suggest that abnormal levels of arachidonic acid metabolites consisting of decreased PGs and increased LTs may mediate salicylate ototoxicity. As with salicylate, quinine ototoxicity appears to be multifactorial in origin. The mechanism includes vasoconstriction and decreases in cochlear blood flow, as measured by laser Doppler flowmetry, motion photographic studies, and histologic studies. Reversible alterations of outer hair cells also appear to play an important role, as demonstrated by histology, electron microscopy, isolated hair cell studies, and cochlear potential evaluations. Unlike with salicylate, however, the role of prostaglandins in quinine ototoxicity has not been clearly demonstrated. Also, one of quinine's principal actions, antagonism of calcium dependent potassium channels, has yet to be investigated for its potential role in ototoxicity. PMID- 8233490 TI - Ototoxic liability of erythromycin and analogues. AB - This article details clinical reports and studies of ototoxicity associated with the administration of erythromycin and its analogues. Suspected mechanisms of ototoxicity also are discussed. Ototoxicity due to erythromycin appears to be clearly dose related. PMID- 8233491 TI - Ototoxicity of vancomycin and analogues. AB - This article details clinical reports and animal studies of ototoxicity associated with vancomycin and its analogues. From these studies, the ototoxicity of these agents is still not clear. In the author's opinion, vancomycin must affect the auditory system in a manner that results in augmentation of the usual ototoxicity of aminoglycoside antibiotics. This postulated effect may manifest as a temporary hearing loss in humans. More studies are needed, however, before a definitive conclusion can be made. PMID- 8233492 TI - Ototoxicity of loop diuretics. AB - The loop diuretics are drugs that increase the excretion of water and electrolytes in the urine by their action on the cells in the loop of Henle. Clinical reports of ototoxicity of these agents are reviewed, and the results of a number of studies in experimental animals are discussed. These drugs can cause either a temporary, or in some cases, a permanent loss of hearing in patients. Animal experiments show that these drugs act on the stria vascularis, producing edema of these tissues and a temporary loss of function, resulting in a decrease of the endocochlear potential. This can result in secondary effects on sound evoked measures of hearing. As new information unfolds about protective agents, it may be possible to preserve hearing and maintain the desired therapeutic effect. PMID- 8233493 TI - Biochemical basis of aminoglycoside ototoxicity. AB - The basis for the development of a rational explanation of aminoglycoside toxicity now appears to exist. The acute effects of these drugs are primarily based on calcium antagonism and block of ion channels. The chronic toxicity requires metabolism, and the expression of tissue-specific toxicity is a balance between synthesis of the toxin and its detoxification. Further investigations into the nature of the toxic metabolite should allow us to combine this information with previously established intracellular actions of aminoglycosides to create a unified hypothesis of action. The ability of glutathione to block toxin formation or to increase detoxification (or both) may have clinical implications for the prevention of aminoglycoside-induced ototoxicity. The clinical use of aminoglycosides has somewhat decreased over the last decade because of the introduction of the less toxic cephalosporins of the third generation and the quinolones, which are effective against Pseudomonas infections. Development of bacterial resistance against aminoglycosides is another factor, although resistance to the cephalosporins is also rapidly becoming a serious problem that eventually will limit their usefulness. Only through a detailed knowledge of the molecular basis of toxicity can we rationally pursue the development of new aminoglycosides with less ototoxic and nephrotoxic potential and devise treatments that will prevent the adverse side effects of these antibiotics. PMID- 8233494 TI - Developmental ototoxicity. AB - The aminoglycoside antibiotics are active bactericidal agents used for the treatment of tuberculosis and gram-negative bacterial infections. Well over 2 million patients per year receive aminoglycosides; however, a major limitation to their use is ototoxicity (cochleotoxicity and vestibulotoxicity), nephrotoxicity, and neuromuscular blockade. Ototoxicity can result from aminoglycoside administration to any patient population, including adults, fetuses, and preterm and full-term neonates. Preterm infants and experimental animals are hypersensitive to aminoglycoside ototoxicity during the period of anatomic and functional maturation of the inner ear. Developing animals, including humans, are also hypersensitive to loop diuretics and especially to the combination of loop diuretics and aminoglycosides. Ototoxicity depends upon several factors in addition to age, including patient population, dose and duration of therapy, genetic and pharmacokinetic factors, and the particular aminoglycoside used. The molecular mechanisms of developmental hypersensitivity have not yet been determined. PMID- 8233495 TI - Current concepts in hair cell regeneration. AB - Hair cell regeneration following various mechanical or chemical insults to the inner ear has become the subject of intense scientific research. This article details current concepts relating to this topic, including descriptions of the hair cell precursor population, the ultrastructural properties of the newly regenerated hair cells, and regeneration of hair cells on the vestibular epithelium. Numerous photomicrographs supplement the text. PMID- 8233496 TI - Audiologic monitoring for ototoxicity. AB - In summary, the clinician has a variety of protocols available for monitoring ototoxicity. Depending on the patient's risk factors and ability to be tested, the protocol for a given patient may vary. A flow chart reflecting some of the possible decisions and options is presented in Figure 1. Certainly, as we learn more about ototoxicity and the advantages and disadvantages of the various test methods, further refinements of patient and test selection will ensue. Ototoxicity is a rapidly expanding and interesting area. Hopefully, the care of patients receiving ototoxic medications will continue to improve and will ultimately prevent, or at least ameliorate, ototoxic hearing loss. PMID- 8233497 TI - Risedronate activity in the fetal and neonatal mouse. AB - Otosclerosis, chronic otitis media with and without cholesteatoma, and Paget's disease of bone are just a few of the many diseases of the ear that exhibit abnormalities of bone modeling and remodeling. These diseases result in chronic infection, vestibular dysfunction, and hearing loss. Bisphosphonates are a promising new class of drugs potentially useful in the treatment of these disorders. Currently used in diseases with high rates of bone turnover (Paget's disease of bone, hypercalcemia of malignancy, and osteoporosis), they have been found to be strong inhibitors of bone resorption. A third generation bisphosphonate, 2-(3-pyrindyl)-hydroxyethylidene bisphosphonate (risedronate) is being investigated for toxicity, increased efficacy, and oral administration. In this study the in vitro and in vivo anti-resorptive activity of risedronate was quantified by measuring calcium release in a neonatal mouse calvarial culture system. This model was used to test direct in vitro effects, in vivo exposure in neonatal mice, and the possible effects of in utero and lacteal exposure. Calcium release activated by parathyroid hormone (PTH) was significantly inhibited when risedronate was only present in the pre-incubation media. When risedronate was administered subcutaneously to neonatal mice it resulted in a significant decrease in PTH-activated calcium release in explanted calvaria in vitro. Transplacental and lactational transfer of biologically effective risedronate was not demonstrated in this study; however, a paradoxic increase in PTH-stimulated calcium release in vitro from calvaria theoretically exposed transplacentally and lacteally was noted. This effect was unexplained by the data. PMID- 8233498 TI - Mucociliary function and endothelins 1, 2, and 3. AB - Endothelins are recently discovered peptides that cause vasoconstriction and bronchoconstriction. The significance to the mucociliary system of endothelins (ET) 1, 2 and 3 in upper as well as lower airways has not yet been clarified. Effects of these active peptides were investigated, combining both in vitro and in vivo rabbit models of mucociliary activity from the maxillary sinus and the trachea. The studies were performed using computerized photometric microscopy. Immunohistologic staining procedures were used to determine the presence of endothelins in sinus and trachea epithelium. Significant effects on mucociliary activity were noted for ET-1, -2, and -3 in vitro as well as in vivo. All endothelins were noted to accelerate mucociliary activity of both sinus and tracheal mucosa. The effects of endothelins were greater in the sinus than in the trachea. In vitro studies using the calcium blocker nifedipine and the cyclooxygenase inhibitor diclofenac indicated that the mechanism of action involves an intermediary prostaglandin pathway but is independent of release of intracellular calcium. These results were confirmed using ET-1 in vivo. Immunohistochemical staining showed endothelin to be present in both maxillary and tracheal epithelium of the rabbit. We conclude that endothelins have significant mucociliary stimulatory effects and that the presence of endothelins in normal mucosa indicates a potentially important role in respiratory homeostasis as well as inflammatory conditions. PMID- 8233499 TI - Control of the mucosal microcirculation in the upper respiratory tract. AB - This study was designed to investigate the regulatory mechanisms of the mucosal microvascular network in the upper respiratory tract. Tracheal mucosal circulation was observed using a specially constructed chamber that allowed direct microscopic visualization of mucosal arterioles. Solutions of increasing hypertonicity (500 and 900 mOsm) applied to the tracheal epithelium resulted in increasing dilation of the underlying mucosal arterioles (p < 0.001). N(omega) nitro-L-arginine methyl ester (L-NAME, 1 mmol/L), a specific inhibitor of nitric oxide synthesis, added to a hypertonic solution inhibited dilation of mucosal arterioles (p < 0.001). Addition of the substrate for nitric oxide synthesis, L arginine (0.6 mmol/L) to the hypertonic solution containing L-NAME resulted in dilation of mucosal arterioles once again. These data demonstrate that nitric oxide is a crucial mediator in the response of mucosal arterioles to the hypertonic stimulus presented to the epithelial surface of the trachea. Further elucidation of the control of the mucosal microcirculation in the upper respiratory tract could be implemented in new treatment for pathologic processes of the upper respiratory tract such as mucosal congestion and edema. PMID- 8233500 TI - A comparison of cocaine vs. lidocaine with oxymetazoline for use in nasal procedures. AB - This study compared the degree of vasoconstriction and anesthesia obtained with cocaine vs. that obtained with a mixture of lidocaine and oxymetazoline (lido/oxy) in healthy, male volunteers. Blood flow was evaluated by laser Doppler flowmetry. Anesthesia was measured with Semmes-Weinstein monofilaments, testing both sensation threshold and pain perception. A greater decrease in blood flow was seen after the administration of lido/oxy than after the administration of cocaine. Pain perception change was not significantly different between treatment groups at 10 minutes after removal of the medication-soaked pledget, but lido/oxy caused a greater decrease in pain perception than cocaine after 50 minutes. No significant difference in sensation threshold change was seen between treatment groups. Lidocaine with oxymetazoline is, therefore, believed to be an effective alternative to cocaine for nasal procedures. PMID- 8233501 TI - Effect of otologic drill noise on ABR thresholds in a guinea pig model. AB - The noise generated by the otologic drill has been implicated as a cause of sensorineural hearing loss after ear surgery. However, clinical studies on this subject are contradictory and difficult to interpret. Therefore a guinea pig model was used to study whether the level of noise generated by the otologic drill can cause threshold shifts in the auditory brainstem response (ABR). The source noise was a recording obtained during a human cadaver mastoidectomy using a microphone and an accelerometer. Ten female Topeka-strain guinea pigs were exposed to the recorded drill noise for a period of 55 minutes. Exposure included both air-conducted energy from a speaker and bone-conducted energy from a bone vibrator applied directly to the skull. ABR threshold measurements were taken pre exposure (baseline), immediately after exposure, and at weekly intervals thereafter for 3 weeks. Three control animals were subjected to the same procedure without the sound exposure. A significant threshold shift (p < 0.0001) was seen for each frequency tested (2, 4, 8, 16, 20, and 32 kHz) immediately after exposure to noise in all experimental animals. Thresholds returned to baseline within 3 weeks. We conclude that the level of noise generated by the otologic drill in mastoid surgery can cause a temporary threshold shift in this guinea pig model. PMID- 8233502 TI - Effect of pentoxifylline [corrected] on myocutaneous flap viability in pigs. AB - Partial necrosis of a skin flap can complicate reconstructive surgery. We performed a double-blinded crossover study to determine if pentoxifylline improves perfusion and survival of a myocutaneous flap. Ten 20-kg pigs were fed pentoxifylline (400 mg three times a day) or placebo for 1 week before and after raising a 5 x 30 cm panniculus carnosus flap on one flank. After 1 week of washout, each pig began the opposite drug treatment and the surgery was repeated on the opposite flank. Immediately after surgery, perfusion dermofluorometry and laser Doppler velocimetry demonstrated a significant increase in the perfusion of pentoxifylline-treated flaps compared to control flaps. On postoperative day 7, the mean area of clinical necrosis was 39.7 +/- 4.7 cm2 on the placebo sides and 30.1 +/- 4.6 cm2 on the pentoxifylline sides (t = 2.21, p < 0.05). We conclude that pentoxifylline improves perfusion and survival of myocutaneous flaps in pigs. Clinical trials appear to be indicated on the basis of the findings of this experiment. PMID- 8233503 TI - Characteristics of cerumen in diabetic patients: a key to understanding malignant external otitis? AB - Malignant externa otitis is a potentially fatal disease in diabetic and other immunocompromised patients. Cerumen contains defense properties that protect the patient against infection. We tested the hypothesis that patients with diabetes mellitus have abnormalities in their cerumen that affect the environment of their external auditory canals and may predispose them to malignant externa otitis. PMID- 8233504 TI - Diuretic and diet effect on Meniere's disease evaluated by the 1985 Committee on Hearing and Equilibrium guidelines. AB - Fifty-four patients, diagnosed with Meniere's disease and treated with diuretics and a low-salt diet, were evaluated retrospectively with the 1985 AAO/HNS Committee on Hearing and Equilibrium (CHE) guidelines for vertigo and hearing changes. The patient data base was also evaluated with other methods that helped determine the effectiveness of the 1985 AAO/HNS CHE guidelines. After 24 months of therapy, vertigo control was complete or substantial in 79% of the patients, limited or insignificant in 19%, and worse in 2% as evaluated by the CHE 1985 guidelines. Hearing improved in 35% of the patients, was unchanged in 29%, was worse in 22%, and could not be classified by CHE guidelines in 14%. Hearing was also evaluated by comparison of individual thresholds before medical therapy, and at 22 and 74 months after the start of medical therapy. We found a stabilization of low- and mid-threshold frequencies, with an average rate of hearing loss approximating 0 dB/yr with 74 months of followup. The results of this preliminary study suggest that diuretics and a low-salt diet may decrease the natural progression of sensorineural hearing loss in patients with Meniere's disease. Compared with other methods of data analysis, the 1985 CHE guidelines lacked sensitivity to evaluate the hearing changes observed. PMID- 8233505 TI - New dimensions for the vital storage of microsurgical free flaps: an experimental approach. AB - A revolutionary circulatory system has been developed to nourish big, free osteomusculocutaneous flaps extracorporally. Thus we will be able to transplant the free flaps to defect areas that have no sufficient vascular situation. In respect to the cold storage of microsurgical free flaps, to date maximal periods of ischaemic tolerance have been considerably exceeded; the maximal period is currently 168 hours. The vitality of the flap is monitored through parameters setting forth the consumption of oxygen together with histology and electron microscopy. The oxygenation of the nutritive medium is achieved through an aqueous phase gas exchange. PMID- 8233506 TI - Are thymus-derived lymphocytes (T cells) defective in the nasopharyngeal and palatine tonsils of children? AB - The present study was conducted to evaluate the response of adenoidal T cells and B cells in the production of immunoglobulins. There appears to be a consistent inability of adenoidal T cells to turn on B cells to mature into immunoglobulin secreting plasma cells. This phenomenon did not appear to be due to suppressor activity of adenoidal T cells because T cells from other sources appeared to effectively result in adenoidal B cell maturation, even in the presence of adenoidal T cells. Both tonsils and adenoids appear to have defective IL-2 production, in response to both mitogens and specific antigens. It is hypothesized that a cytokine(s) may be released in adenoids that downregulate IL 2 production and result in immune suppression in the adenoids of children with recurrent otitis media and chronic sinusitis. PMID- 8233507 TI - The role of gentle ventilation in prevention of subglottic stenosis in the newborn. AB - Prolonged endotracheal intubation has become the standard of care in most neonatal units for maintenance of mechanical ventilation in the presence of respiratory distress. Unfortunately this approach has become associated with significant complications, including acquired subglottic stenosis. We have successfully used nasal continuous positive airway pressure to avoid or decrease the incidence and duration of endotracheal intubation. With use of this technique we have been able to significantly reduce sequelae (i.e., bronchopulmonary dysplasia, chronic lung disease, intraventricular hemorrhage) and have not encountered subglottic stenosis in more than 200 cases. The use of this technique may be of significant value in preventing or reducing the incidence of acquired subglottic stenosis. PMID- 8233508 TI - The safety and efficacy of bedside tracheotomy. AB - Many institutions require that tracheotomies be performed in the operating room. Movement of critically ill patients dependent on multiple life support systems is technically difficult, labor intensive, and potentially dangerous for the patients. Between 1983 and 1992, 1088 tracheotomies were performed on patients ages 1 week to 94 years at the University of Rochester affiliated hospitals on critically ill patients as isolated procedures. The procedure was performed in the Intensive Care Units (ICU) on 996 patients, (92.9%), whereas 92 patients (7.1%) had tracheotomies in the operating room (OR1). An additional 346 tracheotomies took place in the operating room in conjunction with other head and neck procedures (OR2). Incidence of perioperative bleeding (within 48 hours) was 2.3% in the ICU group, 2.1% in the ORI group, and 2.0% in the OR2 group. Incidence of stomal infection was also similar among the three groups at 1.8%, 2.1%, and 1.5%, respectively. Tube dislodgement in all groups was a complication. No statistical differences were noted among the three groups (ICU, OR1, OR2) at the p < 0.01 level. Criteria for performing the tracheotomy in the ICU are delineated and discussed. PMID- 8233509 TI - The retreat of Hemophilus influenzae type B invasive disease: analysis of an immunization program and implications for OTO-HNS. AB - Hemophilus influenzae type b (Hib) is the most common cause of bacterial meningitis among children under 5 years old. Hib is also responsible for other invasive diseases including epiglottitis, cellulitis, sepsis, pneumonia, and osteomyelitis. A child's cumulative risk of systemic Hib disease during the first 5 years of life is approximately 1 in 200. A polysaccharide Hib vaccine was first marketed in 1985, and newer, more effective conjugated vaccines have been licensed since 1987. Immunization schedules have included increasingly younger children. No studies have been published that analyze the effects of a vigorous immunization program on a sample population representative of the United States at large. Records of pediatric patients ages 5 years and younger who were treated for Hib meningitis or epiglottitis (N = 373) at all U.S. Army medical facilities between 1986 and 1991 were reviewed. The combined incidence of these diseases declined by more than 86% in the study group during this period. The largest decrease occurred in infants less than 1 year old, before vaccines were licensed for use in this group. Meanwhile, the number of cases of bacterial meningitis due to other organisms in this cohort remained unchanged. Economic modeling validates the cost-effectiveness of vaccination. The impact of these preliminary trends on health care systems and otolaryngology-head and neck surgery will be significant. Almost two thirds of Hib disease has involved infants under 15 months old, for whom a conjugated vaccine has been available only since October 1990. The change in disease frequency will have substantial bearing on training programs, because management of neurologic sequelae and the emergent airway require the expertise of otolaryngologists. In the face of medical onslaught, Hib invasive disease is in retreat. PMID- 8233510 TI - Effective treatment protocol for posterior epistaxis: a 10-year experience. AB - Posterior epistaxis remains a serious health problem. Modification of a traditional posterior pack using a Foley catheter has proved to be an effective method of treating these sites of severe posterior bleeding. Technological advancements in the form of a PCA infuser and pulse oximetry have allowed better control of packing-associated pain and identification of potential hypoxic complications. PMID- 8233511 TI - The effects of preoperative steroids on tonsillectomy patients. AB - A prospective, randomized, double-blind study to determine the postoperative efficacy of steroids in tonsillectomy was performed in 49 children. A single dose of intravenous dexamethasone or placebo was administered after each child was anesthetized. Postoperatively each child was examined for objective signs of trismus (measured by interincisor distance), temperature elevation, and weight loss, as well as for subjective signs of mouth odor, oral intake, pain, level of activity, and analgesic usage. There were no statistical differences noted in any of the variables compared in the two groups and the complication rates were also similar. PMID- 8233512 TI - Hypersensitivity to latex in health care workers: report of five cases. AB - Concern about transmission of acquired immunodeficiency syndrome and hepatitis has greatly increased the use of latex gloves. Latex allergy is a newly emerging problem with potential life-threatening sequelae among health care personnel. Patients are also at risk. We report on five cases of latex allergy in health care workers who were using latex gloves for variable lengths of time before their allergic reaction. All workers had a significant history of reactions to other allergens, including inhalants and food. Each individual had at least one systemic symptom that was directly attributed to latex exposure. Reactions to latex ranged from severe contact urticaria (all cases), bronchospasm (three cases), angioedema (two cases), and rhinorrhea (one case) to anaphylactic reactions that required immediate attention in three cases. Levels of specific IgE to latex (as measured by in vitro enzyme immunoassay) and total IgE were elevated in all five patients (total IgE > 100 kU/L). Long-term treatment included avoidance of latex, administration of antihistamines, and desensitization to other, nonlatex allergens. Health care workers should have increased awareness of latex allergy. At risk are health care workers with a history of other allergies and those with atopic dermatitis or eczema. In vitro testing may be useful, especially in persons with skin conditions such as eczema or urticaria or for persons with a history of systemic reactions to the latex allergen as reported in these five cases. PMID- 8233513 TI - Double-blind, placebo-controlled trial of rehabilitation for bilateral vestibular hypofunction: preliminary report. AB - Vestibular rehabilitation (VR) is increasingly popular, but few data exist to support enthusiasts' claims of efficacy in improving functional abilities of patients with bilateral vestibular hypofunction (BVH). A double-blind, controlled study of eight subjects (mean, 64 +/- 12 years; seven females, one male) with bilateral vestibular hypofunction was conducted. Subjects in group A received 8 weeks of VR followed by 8 weeks of home VR exercises, whereas those in group B received 8 weeks of control treatment (isometric strengthening exercises) followed by 8 weeks of VR. At the end of 8 weeks, group A walked 8% faster and, during paced gait and stair-climbing, with greater stability, evidenced by a 10% larger maximum moment arm and a 17% decreased double-support duration during gait and stair stance. Group B improved less than 1% during the control treatment. Self-reported Dizziness Handicap Inventory scores did not differ significantly between control and active VR. All subjects improved compared with baseline tests at the 16-week post-test on both functional testing and on the Self-reported Dizziness Handicap Inventory scale. We conclude that in this small sample, VR effectively improved functional, dynamic stability during locomotion, but even strengthening exercises result in self-reported symptomatic improvement. PMID- 8233514 TI - Results of revision stapedectomy for conductive hearing loss. AB - Revision stapedectomy operations performed over a 13-year period (1977 to 1990) for a conductive hearing loss are reviewed in terms of intraoperative findings and hearing results. All operations were performed in a conventional manner without use of laser techniques. A management algorithm based on intraoperative findings is described. Results are compared with previously reported series. The 66 cases include 20 males and 46 females, ranging in age from 8 to 73 years. Mean time between original and revision stapedectomy was 12.5 years. Prostheses encountered at time of revision included wireloop (29), Robinson (18), polyethylene (14), other (3), and two were not found. The most common cause of failure was displacement of the prosthesis. Incus erosion was found in 48% of wireloops, 35% of polyethylene, and only 11% of the Robinson prostheses. Revision resulted in closure of the pure-tone average (PTA) air-bone gap to within 10 dB in 46% and to within 15 dB in 76%. Sensorineural hearing loss (> 10 dB) occurred in 5 cases (7.6%), and a decline in speech discrimination (> 10%) occurred in 17%. Three of four cases requiring drillout had gap closure to within 10 dB. Findings suggest that our management technique produces results comparable to other large reported series. Drillout at the time of revision can be recommended. PMID- 8233515 TI - Detection of largyneal sensory-evoked potentials (LSEPs) in the cat. AB - We previously reported on evoked potentials elicited by electrical stimulation of the superior laryngeal nerve at an appropriate site on the dural surface as a reflection of activities in the brain stem and cortex in anesthetized cats. This evoked potential was called the laryngeal sensory evoked potential (LSEP). In this study we attempted to establish a less invasive procedure for measuring LSEP. The procedures were recording on the scalp using chloride-coated silver disk electrodes and stimulation by insertion of a bipolar platinum hooked wire electrode into the laryngeal mucosa. Evoked potentials could be detected using these less invasive procedures. The response morphologies and relative timing of LSEP components were quite similar for each method in a given cat. However, the amplitudes were slightly lower and the latencies were slightly prolonged with the less invasive techniques. These results suggest that this LSEP method might be applicable to human beings as a noninvasive method for evaluating the function of the laryngeal sensory pathway. PMID- 8233516 TI - Infantile myofibromatosis: a commonly misdiagnosed entity. PMID- 8233517 TI - Herpes simplex esophagitis mimicking a lye burn in a child. PMID- 8233518 TI - Ethmoid cholesterol granuloma. PMID- 8233519 TI - Laryngocele after childbirth. PMID- 8233520 TI - Orbitocranial exposure in the management of infratemporal fossa tumors. PMID- 8233521 TI - Disseminated Pneumocystis carinii infection involving the neck and nasopharynx. AB - In summary, several points should be emphasized: 1. Adenoidal hypertrophy in a person who previously underwent adenoidectomy should lead to the suspicion of HIV infection. 2. P. carinii should be included in the differential diagnosis of adenoidal enlargement in HIV infections. 3. Extrapulmonary calcification should lead to the suspicion of P. carinii infection. PMID- 8233522 TI - Modifications of a nerve integrity monitor useful in skull base surgery. PMID- 8233523 TI - A new stent for endoscopic sinus surgery. PMID- 8233524 TI - Congenital cholesteatoma. PMID- 8233525 TI - Human brain measures of clinical pain: a review. I. Topographic mappings. AB - A comprehensive paper (Parts I and II) has been developed to review the cerebral measures employed in studying the brain neurophysiological activities of clinical pain. Part I focuses on the electro-, magnetic-physiological assessment of clinical pain, and Part II concerns the anatomico-, chemical-physiological assessment of clinical pain. In Part I, these measures include the qualitative inspection of the conventional electroencephalogram, quantitative assessment of brain electrical spectral activity through cortical power spectrum density and coherence analyses, and quantitative averaging of cortical electrical or magnetic activities using brain evoked potentials. The mapping and measurement of these electrical activities and magnetic fields are results of recent advent in computer technology and advanced algorithms. Promises and limitations of these topographic measures in understanding pain in the brain are stated. The next article (Part II) of this paper will review tomographic imaging of pain-related brain activities in regional cerebral flow, the scanning of gross and fine brain structures by computerized axial tomography or magnetic resonance imaging, and the imaging and measurement of brain metabolic changes, energy uptake, and receptor bindings through positron emission tomography or single-photon emission computerized tomography. Molecular chemical transformation by the nuclear magnetic resonance analysis of tissue changes and analgesic-receptor interactions will also be noted. PMID- 8233526 TI - Human brain measures of clinical pain: a review. II. Tomographic imagings. AB - This paper (Parts I and II) reviews the measures employed in studying the brain neurophysiological activities of clinical pain. In Part II, these measures include the imaging and measurement of brain blood flow and hemodynamics in various regions of the brain, the scanning of gross and fine brain structures by computerized axial tomography or magnetic resonance imaging, and the imaging and measurement of brain metabolic changes, energy uptake, and receptors bindings through positron emission tomography or single-photon emission computerized tomography. Molecular chemical transformation by the nuclear magnetic resonance analysis of tissue changes and analgesic-receptor interactions is also noted. Most studies of the cerebral measures of traumatic and pathophysiological pain reported in the literature are concerned with headache. The relationships of brain activities among sensory processes of nociception, subjective experience of pain intensity and quality, emotional reaction, and cognitive coping often are complex and not well elucidated in man. Although significant changes in the cerebral physiological parameters are frequently reported in pain patients, the specificity and sensitivity of these measures as objective markers for human pain, reviewed from Part I and Part II together, has not yet been conclusively defined. Also, normative data basis and criteria for classifying abnormality of these brain measures must be established and their validity and reliability be carefully examined so that they can be confidently applied in diagnosis and management of clinical pain. Nevertheless, advancement on measurement of temporal dynamics in 3-D topographic mapping of cortical activities and source localization modeling, together with tomographic imaging of neurochemical metabolisms in the brain will further our scientific understanding of cerebral pain mechanisms. A window on the brain of human pain is being opened. PMID- 8233527 TI - Neurolytic superior hypogastric plexus block for chronic pelvic pain associated with cancer. AB - Twenty-six patients with extensive gynecologic, colorectal or genitourinary cancer who suffered uncontrolled, incapacitating pelvic pain were enrolled in this study during a 1-year period. All the patients receiving oral opioids who developed poor pain response due to the progression of disease or untoward side effects necessitating other modes of therapy were eligible to participate. Bilateral percutaneous neurolytic superior hypogastric plexus blocks with 10% phenol were performed in every patient, 1 day after receiving successful diagnostic blocks using 0.25% bupivacaine (BUP). All patients reported a visual analog pain score (VAPS) of 10 of 10 before the block. Eighteen patients (69%) had satisfactory pain relief (VAPS < 4 of 10): 15 (57%) after 1 block and 3 (12%) after a second block. The remaining 8 patients (31%) had moderate pain control (VAPS 4-7 of 10) after 2 blocks and received epidural bupivacaine-morphine (BUP MS) therapy with good results. Both groups experienced significant reductions in oral opioid therapy after the neurolytic blocks. No additional blocks were required by patients who had a good response during a follow-up period of 6 months. No complications related to the block were experienced by any patient. In conclusion, neurolytic superior hypogastric plexus block was both effective in relieving pain in 69% of the patients studied (95% confidence interval of 48 85%). Additional neurolytic blocks using higher volumes of the neurolytic agent may be needed in patients with extensive retroperitoneal disease, a group in whom moderate or poor results should be expected. PMID- 8233528 TI - Characteristics of chronic pain patients: factor analysis of the MMPI-2. AB - The Minnesota Multiphasic Personality Inventory (MMPI) has been used extensively in the assessment of chronic pain patients. More recent studies using cluster analytic techniques, investigation of sub-scales, and factor analysis have refined its usefulness. The present study factor-analyzed MMPI-2 data from a sample of 114 chronic pain patients from two different treatment programs. MMPI-2 t scores for the validity and clinical scales, as well as the Harris and Lingoes sub-scales for scales 1, 2 and 3, were used in the analysis. Four interpretable factors emerged and were labeled 'Psychological Dysfunction', 'Interpersonal Isolation', 'Psychomotor Retardation', and 'Physical Dysfunction' to reflect factor loadings. Results are discussed in terms of demonstrating support for using the MMPI-2 factors to refine the assessment of chronic pain patient characteristics. PMID- 8233529 TI - Tyramine conjugation deficit in patients with chronic idiopathic temporomandibular joint and orofacial pain. AB - This study was carried out to explore the value of the tyramine conjugation test, an established trait marker for 'endogenous unipolar depression', in patients with chronic idiopathic temporomandibular joint and orofacial pain. Our results show that the pain patients excrete significantly lower amounts of tyramine sulphate than controls (P < 0.0004). Psychiatric assessment by the structured clinical interview for the diagnosis of mental disorders according to DSM-III-R revealed that 48% of the patients had a history of depression and 10% were currently depressed. However, the never-depressed group of patients had the lowest tyramine sulphate excretion values. These findings suggest that a common biological abnormality underlies the pathogenesis of both chronic idiopathic facial pain and depression. PMID- 8233530 TI - The use of topical guanethidine in the relief of dentine hypersensitivity: a controlled study. AB - After a pilot study had indicated that the topical application of guanethidine might be of value in the treatment of tooth dentine hypersensitivity, a double blind study was undertaken to investigate whether guanethidine was more effective than a distilled water control in alleviating dentinal pain. In 39 adult subjects, who had complained of dentine hypersensitivity, a 1 sec blast of air from a dental 3-in-1 syringe was directed onto sensitive root dentine at an angle of 90 degrees from a distance of 1 cm. The severity of the resulting pain was indicated by the subject on a 10 cm visual analogue scale (VAS). One droplet of either 1% guanethidine monosulphate (Ismelin, Ciba-Geigy) or distilled water, allocated at random, was applied to the dentine for 1 min. The tooth was re tested with the standardised air blast and a second VAS was marked by the subject. In total, 19 patients received guanethidine and 20 received water. VAS scores before treatment in the 2 groups treated with either guanethidine or water (means: 6.5 +/- 2.2 and 6.0 +/- 2.5, respectively) were not significantly different (Mann-Whitney, P = 0.48) whereas the VAS scores in the 2 groups after treatment with guanethidine or water (means: 2.7 +/- 2.4 and 4.8 +/- 2.9, respectively) were significantly different (Mann-Whitney, P < 0.02). It was concluded that guanethidine reduced dentinal hypersensitivity more effectively than distilled water.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233531 TI - Management of terminal cancer pain in Sweden: a nationwide survey. AB - The aim of this nationwide survey was to evaluate the extent of pain problems, the principles of pain management and the use of newer drug delivery techniques in terminal cancer patients. 'Terminal period' was defined as an expected life span of about 2 weeks. Another aim was to evaluate the level of knowledge of physicians and nurses and to assess their educational needs regarding management of cancer pain. A total of 456 questionnaires were sent to chairmen and head nurses of 6 major specialties (228 departments) that treat cancer patients in Sweden. Depending on the specialty, each questionnaire consisted of 50-60 questions. The response rate was high, ranging from 79% (surgeons) to 100% (gynecological oncologists). During a 1-year period 3767 patients (about 10% of total cancer population) were admitted to hospitals due to severe cancer pain. The results show that almost all physicians follow the analgesic 'ladder' principle recommended by WHO, prefer the oral route for morphine administration, prescribe opioid analgesics on a regular 'by-the-clock' basis and in unrestricted doses. However, about 30% of physicians believe that all patients have moderate to severe pain at the time of their death. Up to 78% of physicians and nurses believe that periodic severe pain is common in terminal cancer patients. In general physicians do not evaluate different pain types nor do they use any instruments for measurement of pain intensity. Surprisingly intermittent subcutaneous (s.c.) or intramuscular (i.m.) injections of morphine are preferred to continuous s.c., intravenous (i.v.) infusions or to intermittent s.c. administration through an indwelling butterfly needle. Few physicians routinely prescribe prophylactic laxatives. Antidepressive medication is rarely used. Neuropathic and bone metastasis pain appear to be poorly managed. More than 50% of physicians and nurses admit that they have inadequate knowledge about pain evaluation techniques, newer analgesics and newer drug delivery systems. More aggressive use of opioids administered by more humane routes and better use of adjuvant therapy can be expected to improve pain management for the terminal patient. There is also a need for better physician and nurse education. PMID- 8233532 TI - Development of secondary hyperalgesia following non-painful thermal stimulation of the skin: a psychophysical study in man. AB - A psychophysical study has been carried out in 10 normal human subjects to examine whether conscious perception of pain is necessary for the development of secondary hyperalgesia. Prolonged thermal stimulation of the skin was applied to the subjects at intensities known to evoke discharges in polymodal nociceptors but insufficient to evoke pain sensations. During this stimulation the development of punctate and of stroking hyperalgesia was examined as was the presence of a skin flare indicative of nociceptor activation. All subjects developed a flare and an area of hyperalgesia following the application of the non-painful heat stimulus. The first change observed in the subjects was the appearance of an area of hyperalgesia to punctate stimuli, followed by flare and by stroking hyperalgesia. The onset of pain was always reported sometime after these events. Statistical analysis of these data for all subjects showed a highly significant difference between the time of onset of pain and the time of onset of any of the other 3 phenomena. Significant differences were also observed between the onset of punctate hyperalgesia and the onsets of flare and of stroking hyperalgesia. No difference was observed between the onset of flare and of stroking hyperalgesia. These results show that cutaneous hyperalgesia can be evoked in normal human subjects by prolonged thermal stimulation of the skin at temperatures that are not perceived as painful. The development of a flare in all subjects simultaneously with stroking hyperalgesia but before the perception of pain suggests that activation of nociceptors is necessary for the hyperalgesia to occur.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233533 TI - Interactions between sympathetic vasoconstrictor outflow and C nociceptor-induced antidromic vasodilatation. AB - The interaction between orthodromic sympathetic vasoconstrictor and antidromic vasodilator effects of C-fiber stimulation was studied in normal human volunteers. Excitation of C fibers was achieved through administration of intermittent painful intraneural microstimulation to upper limb nerves, while recording cutaneous thermal emission profiles in the projected fields of cutaneous sensation. During initial stimulation, when both systems are co activated, skin temperature decreased. Temperature increased after termination of the stimulus. Eventually, temperature decreased again upon renewed stimulation. Thus, sympathetic vasoconstriction was found to override the antidromic vasodilator effect induced by stimulation of C nociceptors. When interpreting abnormal deviations of skin temperature in patients with peripheral nerve disorders, the dynamic interplay between opposing vasomotor effects driven by sympathetic and somatic systems must be taken into consideration. PMID- 8233534 TI - Diclofenac evaluated in a human experimental model of central pain. AB - The putative central analgesic activity of diclofenac was investigated in a human experimental pain model using intraneural electrical stimulation in the median nerve. Since pain is induced proximal to the peripheral nociceptors, the model can be used to test central analgesic properties of i.a. pharmacological interventions performed during series of repeated stimulations. A single intravenous dose of 50 mg diclofenac or saline was administered during an ongoing series of painful intraneural stimulations in a double-blind cross-over study in 10 healthy volunteers. Neither diclofenac nor saline caused any significant change in the level of pain experienced during stimulation. Thus, no central analgesic effect of diclofenac could be demonstrated in this model. The stability of individual visual analogue scale (VAS) scores throughout the experimental sessions, also after administration of the potent peripheral analgesic agent diclofenac, underlines the validity of intraneural stimulation as a central pain model in humans. PMID- 8233535 TI - Increased in vivo release of calcitonin gene-related peptide-like material from the spinal cord in arthritic rats. AB - Possible alterations in spinal systems containing calcitonin gene-related peptide (CGRP) due to polyarthritis were assessed in rats 3-4 weeks after an intradermal injection of Freund's adjuvant in the low back. The tissue levels of CGRP-like material (CGRPLM) were approximately 50% higher in the dorsal zone of the spinal cord and dorsal root ganglia at both the cervical and lumbar (but not thoracic) segments in polyarthritic rats than in age-paired control animals. In addition the rate of the spinal release of CGRPLM determined through an intrathecal perfusion procedure in halothane-anaesthetized animals was approximately 15-fold higher in polyarthritic rats than in controls. The blockade of mu-opioid receptors by intrathecal perfusion with 10 microM naloxone produced a larger increase in the spontaneous CGRPLM outflow in polyarthritic rats than in age paired controls. Furthermore, the stimulation of mu-opioid receptors by intrathecal perfusion with 10 microM DAGO significantly inhibited the spinal outflow of CGRPLM only in polyarthritic rats. These data indicate that CGRP containing primary afferent fibres are markedly activated in chronic suffering polyarthritic rats. This activation occurs in spite of an increased tonic inhibitory control by endogenous opioids acting at mu receptors. PMID- 8233536 TI - On the nature of neck pain, discography and cervical zygapophysial joint blocks. AB - To determine the prevalence of disc pain and zygapophysial joint pain occurring simultaneously in the same segment of the neck, 56 patients with post-traumatic neck pain underwent both provocation discography and cervical zygapophysial joint blocks. Both a symptomatic disc and a symptomatic zygapophysial joint were identified in the same segment in 41% of the patients. Discs alone were symptomatic in only 20% of the sample. Zygapophysial joints were symptomatic but discs asymptomatic in 23%. Only 17% of the patients had neither a symptomatic disc nor a symptomatic zygapophysial joint at the segments studied. These observations indicate that the investigation of neck pain by discography alone or by zygapophysial blocks alone constitutes an inadequate approach to neck pain which fails to identify the majority of patients whose symptoms stem from multiple elements in the 3-joint complexes of the neck. PMID- 8233537 TI - Ketamine hydrochloride in the treatment of phantom limb pain. AB - Three cases are described in which phantom limb pain was successfully treated with ketamine hydrochloride. Possible mechanisms for this observation are suggested. PMID- 8233538 TI - The 'significance' of statistical significance. PMID- 8233539 TI - Significance, non-significance--who has found significance? PMID- 8233541 TI - Vigilance in defense of animal welfare. International Association for the Study of Pain. PMID- 8233540 TI - The role of conditioning in the placebo response: a comment on Voudouris et al., PAIN, 43 (1990) 121-128. PMID- 8233542 TI - Nociception from skeletal muscle in relation to clinical muscle pain. PMID- 8233543 TI - Spinal nitric oxide synthesis inhibition blocks NMDA-induced thermal hyperalgesia and produces antinociception in the formalin test in rats. AB - To assess the possible role of spinal nitric oxide (NO) synthesis in nociceptive processing, we examined the effect of intrathecal (i.t.) injection of arginine analogs that act as alternate substrates for NO synthase and thus inhibit NO production. NG-nitro-L-arginine ester (r-NAME) and NG-monomethyl-L-arginine (L NMMA) produced a dose-dependent, stereospecific inhibition of the second phase (10-60 min; ED50, 135 and 246 nmol) of the formalin test with minimal effect on the first phase (0-9 min; ED50 > 1.1 mumol). The inhibitory action of L-NAME was dose-dependently reversed by i.t. L-arginine (ID50, 4.9 mumol) but not by D arginine (ID50 > 14 mumol). The suppression of the second-phase formalin response by L-NAME was similar whether administered before or after formalin injection into the rat paw. Spinal administration of L-NAME (370 nmol), but not D-NAME (3.7 mumol), also blocked thermal hyperalgesia induced by i.t. injection of N-methyl-D aspartate (NMDA; 6.8 nmol). The effect of L-NAME was reversed by L-arginine (4.7 mumol) but not with D-arginine (14 mumol). None of the compounds, L-NAME, D-NAME or L-arginine, when injected alone, had any effect on normal thermal response latencies or on the 52.5 degrees C hot plate. These studies indicate that modulation of spinal NO synthesis can diminish the facilitated processing of afferent activity which is induced by a continued afferent barrage (second phase of the formalin test). This hyperalgesic component appears initiated by the activation of a spinal NMDA receptor that, through the generation of NO, leads to the observed augmented processing of afferent input and the associated hyperalgesic component of the subsequent pain behavior. PMID- 8233544 TI - The NMDA antagonist Memantine blocks pain behavior in a rat model of formalin induced facial pain. AB - Recent studies have provided evidence that excitatory amino acid antagonists can exert analgesic effects in animals. These studies, however, have focused primarily on phasic pain or hyperalgesia rather than tonic pain. The present study evaluates the effects of systemic administration of Memantine (1-amino-3,5 dimethyl-adamantane), a clinically used N-methyl-D-aspartate (NMDA) receptor antagonist, on formalin-induced phasic and tonic pain behavior in the rat. Memantine (2.5, 5.0, 10.0 and 20.0 mg/kg) or normal saline was injected i.p. 1 h prior to a s.c. injection of formalin (5%, 50 microliters) into the vibrissal pad of adult rats (n = 5/group). Pain behavior was measured by the number of seconds of formalin-induced face grooming during a 42-min post-injection observation period. Saline-injected animals displayed a biphasic face-grooming response, consisting of an early, phasic phase (0-6 min) and a delayed, prolonged tonic phase (12-42 min). Memantine at doses of 2.5-10 mg/kg produced a significant dose related inhibition of the second phase (65-93%) and a much smaller inhibition of the first phase (up to 52%). A higher dose (20 mg/kg) further inhibited both phases but also produced other motor effects (increased exploratory and decreased freezing behavior, hind-paw weakness and gait ataxia) which were not observed at the lower doses. These results suggest that the NMDA receptor antagonist Memantine can block formalin-induced tonic and, to a lesser extent, phasic pain, at doses that do not alter observed motor behaviors. PMID- 8233545 TI - Spinal cord morphology and antinociception after chronic intrathecal administration of excitatory amino acid antagonists in the rat. AB - Drugs that antagonize the action of excitatory amino acids on the NMDA receptor in the spinal cord are of interest in pain treatment. Before such drugs can be applied clinically, their potential toxicity should be studied. This study was performed in rats in order to reveal possible neurotoxicologic side effects following chronic intrathecal (i.t.) application of two NMDA receptor antagonists: 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) and kynurenic acid (KYN). Rats equipped with i.t. catheters were injected twice a day for 2 weeks with saline, 2 nmol (0.5 micrograms) CPP or 210 nmol (40 micrograms) KYN, where the doses of CPP and KYN were chosen on the basis of similar analgesic effects after one administration. Antinociception was tested daily using the tail flick and hot-plate tests. The antinociceptive effect was similar in CPP- and KYN treated rats on days 1 and 2. The effect of CPP decreased during the following days, whereas that of KYN persisted for the 12-day testing period. The spinal cord was then removed and prepared for light and electron microscopic examination, and a morphometric method using an unbiased stereological estimator of cell number and cell volume was applied as a sensitive variable of spinal cord neurotoxicity. Morphologic and ultrastructural analyses of the spinal cord segment adjacent to the tip of the catheter showed normal appearance with no differences between the groups. Furthermore, no differences in cell number or cell volume in the dorsal horn were found between the groups. In conclusion, chronic i.t. administration of pharmacologically active doses of CPP and KYN in rats did not produce neurotoxic effects in the spinal cord.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233546 TI - A psychosocial and behavioral comparison of reflex sympathetic dystrophy, low back pain, and headache patients. AB - Based primarily on anecdotal evidence, patients with reflex sympathetic dystrophy (RSD) have often been suspected of having a high degree of psychosocial disturbance prior to the onset of symptoms as well as in reaction to the disorder. In the present study, patients presenting to a pain center with RSD were compared to patients with low back (LBP) and headache pain (HAP) on a variety of self-reported demographic, behavioral, pain, and mood measures. Typical of most patients experiencing chronic pain, all three groups demonstrated elevations indicative of pain, emotional distress, and behavioral disturbance. However, although the RSD patient group reported the highest level of pain intensity, the most employment disruption, and contained the highest percentage of patients receiving financial compensation, this same group paradoxically reported less emotional distress on the Symptom Checklist-90R than did LBP and HAP patients. This paradox may be due to the lesser chronicity of the RSD patients as well as to their apparently experiencing a more sympathetic response from doctors, employers, and insurance carriers than their LBP and HAP counterparts. On balance, the present data do not support the hypothesis the RSD patients, relative to other pain patients, are uniquely disturbed in psychosocial functioning. PMID- 8233547 TI - How frequent is anesthesia dolorosa following spinal posterior rhizotomy? A retrospective analysis of fifteen patients. AB - Anesthesia dolorosa has been considered an infrequent complication of spinal posterior rhizotomy. We reviewed the data of all patients who underwent rhizotomy between 1962 and 1972 (15 cases). Thirteen were affected by cancer and 2 by non neoplastic conditions. Eight developed a typical deafferentation pain (i.e., anesthesia dolorosa) (53%) while 3 who were found to have a brain (frontal 2; parietal 1) metastasis did not. Anesthesia dolorosa developed 1.5-8 months after rhizotomy. We conclude that anesthesia dolorosa following rhizotomy is more frequent than usually stated and that rhizotomy should be restricted to patients with a less than 3-month life expectancy. PMID- 8233548 TI - Nurses' assessment of postoperative patients' pain. AB - This study examined nurses' assessments of pain in postoperative patients. Subjects were 119 registered nurses and 119 abdominal surgery patients at two community and two university hospitals. Patients completed a visual analogue scale (VAS) when they indicated the presence of pain. Simultaneously, the nurse with the primary responsibility for the patient's care completed a VAS to indicate an assessment of the patient's pain. The results indicated nurses' assessments were modestly, but significantly, correlated with their patients' pain. The majority of the nurses underassessed their patients' pain. Further inspection of the data, however, revealed that nurses' assessments were related to the degree of pain experienced by patients. Nurses underassessed more severe pain and overassessed mild pain. Nurses' accuracy in assessments were related neither to patients' age nor to the length of time since surgery. Implications for clinical practice and suggestions for further research are discussed. PMID- 8233549 TI - Failure hurts: the effects of stress due to difficult tasks and failure feedback on pain report. AB - Two experiments were conducted to investigate the effects of task difficulty and perceived success/failure on pain report. The first experiment found that difficult reading comprehension problems led to an increase in pain report. The second experiment found that task difficulty per se may not have accounted for the effects, but that perceived failure of the more difficult task led to increased pain report. Social and theoretical implications are discussed. Failure may lead to negative affectivity, which can increase the report of painful stimuli. PMID- 8233550 TI - Polarised views on treating neurogenic pain. AB - This study aimed to identify areas of disagreement in the management of neurogenic pain. A short questionnaire was mailed to 179 consultants with an interest in chronic pain (response rate 89%). The questionnaire listed 11 specific conditions involving nerve pain (e.g., post-herpetic neuralgia, causalgia) together with 11 treatments (e.g., antidepressants, neurectomy). Consultants were asked to rate the use of each treatment for each condition as 'appropriate', 'no value or positively harmful' or 'no opinion'. Much disagreement emerged about the value of each therapy for each condition: in almost every instance at least some consultants disagreed with the majority view. The dissenting minority was greater than 20% of those who gave an opinion for 48 of the 121 applications of therapy asked about. The appropriateness of treatments for trigeminal neuralgia, amputation stump pain and phantom pain was most often in dispute and there was little consensus on the value of nerve blocks. There were a few areas of near agreement. Antidepressants and anticonvulsants were mostly identified as appropriate for all the conditions listed and there was some agreement that strong opioids and the neuroablative techniques were appropriate for cancer pressure or infiltration of nerves but, with a few exceptions, of no value for all other neurogenic pain conditions. Divergence of views about treatments may indicate a lack of credible evidence on the value of therapies or a lack of professional knowledge. Where published evidence is clear, the consequences for patients may be under-use of useful therapies or potential iatrogenic harm. PMID- 8233551 TI - Memory for pain and affect in chronic pain patients. AB - Forty chronic pain patients recorded their levels of sensory and affective pain, and anxiety and depression at the beginning of a 6-week pain management program. At the completion of the program, they again recorded their levels and also rated their memory for each dimension at the beginning of the program. Results indicated that patients reported only marginal improvement after the program. Patients who reported increased pain or depression over the course of the study overestimated their memory for their initial pain or depression. Results are discussed in terms of the influence of subjective changes on pain memory distortion and the need to investigate the interaction between patients' mood and autobiographical memory. PMID- 8233552 TI - A controlled study of the effects of an early intervention on acute musculoskeletal pain problems. AB - Current conceptions of chronic pain clearly suggest that proper care at the acute stage should prevent the development of chronic problems. Patients (198) seeking help for acute musculoskeletal pain (MSP), e.g., back and neck pain participated in two studies of the effects of an Early Active intervention which underscored 'well' behavior and function compared to a Treatment as usual control group. The quantity of the Early Active treatment was a median of 1 doctor's appointment and 3 meetings with a physical therapist. Study I concerned patients with a prior history of sick-listing for MSP, while study II involved patients with no prior history of MSP. Treatment satisfaction, pain experience, activities and sickness absenteeism were assessed before, after and at a 12-month follow-up. In study I (patients with a history of MSP), the results showed significant improvements for both groups, but virtually no differences between the groups. Similarly, in study II (no history of MSP) both groups demonstrated significant improvements, e.g., for pain intensity and activity levels. However, the Early Active treatment resulted in significantly less sick-listing relative to the control group. Moreover, the risk of developing chronic (> 200 sick days) pain was 8 times lower for the Early Activation group. This investigation shows that relatively simple changes in treatment result in reduced sickness absenteeism for 'first-time' sufferers only. Consequently, the content and timing of treatment for pain appear to be crucial. Properly administered early intervention may therefore decrease sick leave and prevent chronic problems, thus saving considerable resources. PMID- 8233553 TI - Comments on topical clonidine for relief from allodynia. PMID- 8233554 TI - Clonidine: effects on pre- vs. post-junctional adrenoceptors. PMID- 8233555 TI - Comments on Coggeshall et al., PAIN, 52 (1992) 233-242. PMID- 8233556 TI - Central neuroplasticity. PMID- 8233557 TI - Comments on Gracely et al., "Painful neuropathy: altered central processing maintained dynamically by peripheral input" (PAIN, 51 (1992) 175-194) PMID- 8233558 TI - Stage and isotype specific immune responses in a rat model of filariasis. AB - Inbred PVG rats infected with 100 Brugia pahangi infective larvae (L3) divide into rats that develop microfilaraemic infection (Mf+), and those that remain microfilaria negative (Mf-). All rats had high levels of specific IgG to adult worm and L3 extracts, however, after the onset of microfilaraemia, Mf+ rats had significantly higher levels of IgG to Mf extract. Mf+ rats recognized several antigenic components of each developmental extract that were not responded to by Mf- rats. In particular Mf+ rats recognized a triplet of proteins of 61-67 kD in microfilarial extract from day 74 post infection onwards. This indicated that these proteins were stage specific to Mf and Mf- rats had not been exposed to Mf rather than Mf absence being due to a protective antibody response. Analysis of the immunoglobulin isotype usage during infection revealed that each isotype was independent both in its period of induction and the developmental stage to which it responded. The predominant isotypes responding throughout infection were IgG1, IgG2a and IgM. Specific IgG2b and IgG2c were elevated early in infection but after the onset of microfilaraemia antibody of these subclasses was suppressed. The antigenic profiles recognized on immunoblots by IgG1, IgG2a and IgM were very similar. PMID- 8233559 TI - Ultrastructural localization of major hydatid fluid antigens in brood capsules and protoscoleces of Echinococcus granulosus of human origin. AB - Monospecific rabbit antisera obtained through experimental immunization with previously purified proteins were used in the ultrastructural localization of two hydatid fluid antigens, in brood capsules and protoscoleces of Echinococcus granulosus of human origin. The antigen-antibody reaction was revealed by a colloidal gold based method. Reaction was evident in the connective region of the germinal membrane and in the parenchyma of the protoscoleces. Both antigen 5 and antigen B were located in the interstitial material between the parenchymal cells and precisely associated with disorganized areas. The brood capsule wall and the brood capsule contents, the tegument of the protoscoleces, the parenchymal cells, the muscle cells, the calcareous corpuscles and the hooks did not contain antigen 5 or antigen B. Label was not observed in the lumen of the collecting ducts or in the flame cells, although antigen 5 was evident in the periluminal cytoplasm. The origin of the antigens and their release are discussed. PMID- 8233560 TI - Type I hypersensitivity reactions in intestinal mucosae from rats infected with Fasciola hepatica. AB - Type I hypersensitivity reactions in the intestinal tract of sensitized animals may contribute to resistance to reinfection with Fasciola hepatica. Colonic mucosae isolated from previously infected rats were voltage clamped in Ussing chambers. Antigen was prepared as a crude homogenate from adult liver fluke. Assay of serum antibodies against fluke antigen confirmed sensitization. Antigen challenge evoked a rapid onset, transient inward current in sensitized but not in control preparations. Chloride secretion accounted for at least part of the response since the loop diuretic bumetanide reduced the effect of antigen by 61%. Anti-rat IgE mimicked the response to antigen and desensitized tissues to subsequent antigen challenge. Local synthesis of eicosanoids may mediate the response to antigen since the cyclo-oxygenase inhibitor piroxicam reduced the response by 76%. In contrast, mepyramine which is a histamine receptor antagonist did not alter the ion transport response evoked by antigen. Tetrodotoxin reduced the response to antigen by 53% implicating intrinsic neurons within the lamina propria as effector cells in the responses of this tissue to antigen. We propose that antigen stimulation of electrogenic chloride movement and consequent fluid secretion in vivo may contribute to a local effector mechanism in prevention of reinfection of previously sensitized hosts. PMID- 8233561 TI - Antibody responses to a 33 kDa cysteine protease of Trypanosoma congolense: relationship to 'trypanotolerance' in cattle. AB - A cysteine protease of Trypanosoma congolense (congopain) elicited IgG1 antibodies in those cattle which exhibited a degree of resistance to disease during experimental infections (Authie et al. 1992, 1993). The aim of the present study was to investigate further the association between anti-congopain antibodies and resistance to trypanosomiasis, and to provide a lead into the mechanisms responsible for the differential responses to congopain in cattle. Isotype characteristics and kinetics of the antibody response to congopain were studied in three N'Dama (trypanoresistant) and three Boran (susceptible) cattle during primary infection with T. congolense ILNat 3.1. In both groups an IgM response to congopain was elicited, thus demonstrating that congopain is antigenic in both types of cattle. Most of the IgM appeared to be incorporated into immune complexes. IgG was detected as free antibody; IgG1 but not IgG2 was detected. All three N'Dama, but none of the three Boran cattle, mounted a significant IgG response to congopain. Sera from 70 primary-infected cattle belonging to five breeds of differing susceptibility were tested for their reactivity to congopain. High levels of IgG to congopain were observed in the two trypanotolerant breeds, whereas the three susceptible breeds had lower levels of these antibodies. Crosses between N'Dama and Boran cattle, which exhibit an intermediate susceptibility, had intermediate levels of antibodies. Thus, the results from experimental infections confirmed our initial observations. However, under natural tsetse challenge, repeated infections and trypanocidal treatments in Zebu cattle stimulated as high anti-congopain antibody levels as in non treated trypanotolerant taurine cattle. PMID- 8233562 TI - Comparison of the effects of immune killing mechanisms on Trypanosoma brucei parasites of slender and stumpy morphology. AB - Trypanosoma brucei slender forms predominate over stumpy forms as the parasite population grows but at the peak of a parasitaemic wave and during remission of infection stumpy forms predominate. To determine whether this change in predominance might be caused by selective killing of slender forms, the fates of slender and stumpy form trypanosomes in two in vitro assays of immune-mediated killing were compared. Parasite populations in which > 90% of cells were of slender morphology were observed to be killed by antibody-dependent complement mediated lysis approximately five times faster than populations in which < 15% of cells were slender and most were of intermediate or stumpy morphology. Quantification of the relationship between the proportion of slender forms in the population and the rate of lysis indicated that slender forms were killed approximately 7.3 times faster than other forms. In an opsonization assay, no differences were observed between slender and stumpy forms in the extent to which they attached to macrophages in an antibody-dependent manner. These results suggest that the change in proportions of slender and stumpy forms at the peak of a parasitaemic wave results from slender forms being more susceptible to complement-mediated killing as the antibody response develops. PMID- 8233563 TI - Salivary gland material from the sand fly Lutzomyia longipalpis has an inhibitory effect on macrophage function in vitro. AB - Previous work from our laboratory demonstrated that the infectivity of the protozoan parasite Leishmania major was enhanced in mice if the infecting inoculum contained salivary gland lysates from the sand fly vector Lutzomyia longipalpis. The present study was designed to address the hypothesis that sand fly salivary gland material may function by inhibiting the host immune response. Results indicated that sand fly saliva inhibited the ability of macrophages to present leishmanial antigens to parasite-specific T cells. PMID- 8233564 TI - Effect of macrophage infection by Leishmania on the proliferation of an antigen specific T-cell line, TPB1, to a non-parasite antigen. AB - The ability of Leishmania mexicana amazonensis to inhibit antigen specific T-cell proliferation against a non-parasite polypeptide antigen, poly(LTyr, LGlu) poly(DLAla)--poly(LLys), was examined. Infection of mouse peritoneal macrophages by promastigotes blocked the proliferation of the T-cell line, TPB1. This effect was correlated with the level of parasite infection, and the timing of macrophage infection and antigen addition. Peritoneal macrophages from both BALB/b and C57BL/6 mice showed reduced ability to serve as antigen presenting cells. PMID- 8233565 TI - Multivariate chemosystematics demonstrate two groups of Actinobacillus actinomycetemcomitans strains. AB - Chemical analysis by us has indicated that Actinobacillus actinomycetemcomitans is not a homogeneous species. The present study used chemometric methods and a multitude of chemical characters to examine this further. Strains were characterized by cell sugar and fatty acid contents, lysis kinetics during EDTA and EDTA plus lysozyme exposure, methylene blue reduction, and API ZYM enzymatic assessment of whole cells and outer membrane vesicles/fragments. In total, 41 quantitative variables were analyzed from each of 9 strains and treated with principal component analysis and soft independent modeling of class analogy. These methods divided A. actinomycetemcomitans into 2 strain groups. One group contained ATCC 33384, ATCC 29522, FDC 2112 and FDC 2043; the other comprised ATCC 29524, ATCC 29523, FDC 2097, FDC 511 and FDC Y4. With an F-test, the groups (classes) of A. actinomycetemcomitans strains could be distinguished at 95% confidence limits. Both groups were distinct from members of the genera Haemophilus and Pasteurella (Haemophilus aphrophilus, Haemophilus paraphrophilus, Haemophilus influenzae, Pasteurella multocida and Pasteurella haemolytica). PMID- 8233566 TI - Killing of Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus by human polymorphonuclear leukocytes in serum and saliva. AB - The ability of polymorphonuclear leukocytes from human peripheral blood to kill Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus was examined with fresh isolates and laboratory strains from each species (5 strains within each group) under different conditions. Bacterial cells were mixed with a polymorphonuclear leukocyte suspension in the presence of either active serum or heat-inactivated serum or active serum together with sterile-filtered saliva. Surviving bacteria were determined by counting the number of bacterial colony forming units in the mixtures after a 60-min incubation at 37 degrees C. Mixtures without polymorphonuclear leukocytes served as controls for the evaluation of the degree of killing of the bacteria. In general, A. actinomycetemcomitans resisted phagocytic killing to a greater extent than H. aphrophilus, and the killing of the former species mainly depended on the presence of heat-labile serum components, probably complement factors. Laboratory strains of A. actinomycetemcomitans were more easily killed than fresh isolates. The presence of saliva in the reaction mixtures decreased the degree of killing. However, strain-dependent variations in the killing were found under either condition. The leukotoxic activity of A. actinomycetemcomitans strains, determined by a [51Cr] release assay, was not correlated with the resistance of these strains to the phagocytic killing. The results point out a strain-dependent difference in the ability of A. actinomycetemcomitans to evade the inflammatory response associated with polymorphonuclear leukocytes. This difference may constitute a potential virulence factor for this periodontopathogen. Furthermore, the leukotoxicity of the strains is not the main determinant that modifies the interaction of A. actinomycetemcomitans with human neutrophils. PMID- 8233567 TI - Analysis of the genetic control of antibody response to Actinobacillus actinomycetemcomitans by immunoblotting inbred strains of mice. AB - Genetic regulation of the immune response may be involved in the onset and progression of an early-onset type of periodontitis. We analyzed the genetic control of the primary antibody response to Actinobacillus actinomycetemcomitans in inbred strains of mice using an immunoblot technique. Mice of 5 independent inbred strains, 6 H-2 congenic strains and 4 B10 intra-H-2 recombinant strains were immunized with sonicated extracts of A. actinomycetemcomitans. On the seventh day their sera were examined for reactivity to the antigenic components of this organism. Western blot analysis clearly distinguished 2 different groups of antigens, one consisting of common antigens (molecular weights, 28, 34, 36 and 40 kDa) that reacted with sera from all strains and one consisting of specific antigens (molecular weights 31, 65 and 69 kDa) that reacted only with sera from distinct strains. Blot analysis of sera from H-2 congenic strains demonstrated that the reactivity to the second group of antigens was regulated by the H-2 complex. In B10 intra-H-2 recombinant strains, only the I-Ab allotype strains produced immunoglobulin G antibody that reacted to the 65 kDa antigen. This evidence indicates that the primary immune response to the A. actinomycetemcomitans antigen with a molecular weight of 65 kDa is controlled by genes in the I-A subregion of the H-2 complex. This 65 kDa antigen was also highly reactive with some human sera from early-onset periodontitis patients. Further analysis of this antigen is required. PMID- 8233568 TI - Serum immunoglobulin G antibodies to Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Streptococcus sanguis during experimental gingivitis in young adults. AB - Twenty-eight young, healthy adults completed an experimental gingivitis study in which blood and clinical recordings were obtained at baseline; after a 4-week period of thorough oral hygiene; after a subsequent 3-week period of plaque accumulation; and after another 2 weeks of thorough oral hygiene. Serum immunoglobulin G antibodies against whole cells of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Streptococcus sanguis were determined using enzyme-linked immunosorbent assay. Mean serum immunoglobulin G antibody levels to P. intermedia, F. nucleatum and S. sanguis remained essentially constant during the experiment, whereas the immunoglobulin G antibodies to P. gingivalis declined during the initial period of oral hygiene and the subsequent period of plaque accumulation to an average of 84.5% of the baseline value. This reduction could be attributed to the people who developed marked gingival inflammation during the period of plaque accumulation, indicating that the systemic host response may be associated with local tissue responses to variations in oral hygiene. These people were, however, also characterized by higher initial serum immunoglobulin G responses to P. gingivalis than people who developed less pronounced gingival inflammation during the experiment. The variability and individuality noted in the host response to potential pathogens have important implications for attempts to use such measures for establishing a diagnosis or prognosis for the individual patient. PMID- 8233569 TI - Immunochemical characterization of the carbohydrate antigens of serotype c/Lancefield group C "Streptococcus milleri". AB - Carbohydrate antigens of the serotype c/Lancefield group C "Streptococcus milleri" were extracted by autoclaving whole cells of the type c reference strain K51Y. The type c and group C antigen molecules were separated and partially purified by a DEAE-Sephadex A-25 column chromatography followed by Sephadex G-100 gel filtration. The purified type c antigen and group C antigen were homogeneous in the double diffusion and in the immunoelectrophoresis. The type c antigen was composed principally of glycerol, rhamnose, glucose and N-acetylglucosamine in a molar ratio of 0.22:0.27:1.00:0.48. The quantitative precipitin inhibition test indicated that N-acetylglucosamine played a major role in immunodeterminant structure. Thus, the type c antigen of "S. milleri" is a new carbohydrate type antigen and is immunochemically different from the Ottens-type antigen III found occasionally in group C streptococci. In contrast, the group C antigen preparation contained a high proportion of N-acetylgalactosamine in addition to glycerol, rhamnose, glucose and N-acetylglucosamine, and the N acetylgalactosamine residue was involved in the immunodominant epitope, being in good agreement with the previously proposed chemical structure of the group antigen. N-acetylgalactosamine was detected in the autoclaved extracts of a nontypeable/group C strain but not of a type c/ungroupable strain. PMID- 8233570 TI - Branched-chain amino acid transport in Streptococcus mutans Ingbritt. AB - Leucine transport in glucose-energized cells of Streptococcus mutans exhibited Michaelis-Menten-type kinetics at low extracellular concentrations, with a K1 of 15.3 microM and a Vmax of 6.1 nmol/mg dry weight/min. At high extracellular leucine concentrations, the transmembrane diffusion of leucine was not saturable, indicating that passive diffusion becomes a significant mechanism of leucine transmembrane movement under these conditions. The proton motive force (PMF) was measured in glucose-energized cells of S. mutans and was found to have a maximum value of 126 mV at an extracellular pH (pH0) of 5.0; this decreased to 45 mV at pH0 8.0. The intracellular accumulation of leucine was significantly correlated with the magnitude of the PMF. The addition of excess isoleucine or valine caused a marked decrease in the leucine transport rate. Maximal rates of leucine transport occurred at pH0 6.0, and the rate of leucine transport was independent of the growth medium. The results suggest that there is a PMF-driven, branched chain amino acid carrier in S. mutans with a proton: substrate stoichiometry of 1. PMID- 8233571 TI - In vitro antimicrobial susceptibility to combinations of drugs on bacteria from carious and endodontic lesions of human deciduous teeth. AB - The purpose of this study was to clarify the antibacterial efficacy of mixed antibacterial drugs on bacteria of carious and endodontic lesions of human deciduous teeth in vitro. The antibacterial drugs used in this study were mixtures of ciprofloxacin, metronidazole, plus a third antibiotic: amoxicillin, cefaclor, cefroxadine, fosfomycin or rokitamycin. Samples taken from carious dentin (17 cases) and infected pulpal tissues (14 cases) were cultured on control plates and plates containing the mixed drugs. No bacteria were recovered in the presence of any combination of the mixture of the drugs (100 micrograms each/ml), and the bacterial growth occurred on control plates (10(1) to 10(7) colony forming units), indicating that the mixed drugs inhibit the growth of bacteria in the samples. When carious and endodontic lesions on split surfaces of freshly extracted teeth were covered overnight with alpha-tricalcium phosphate cement containing a mixture of ciprofloxacin, metronidazole and cefaclor (1% each; 5 cases), no bacteria were recovered from the lesions. No bacteria were recovered from carious and endodontic lesions when these lesions were immersed in a solution of the mixture (200 micrograms each/ml; 5 cases). These findings indicate that carious and endodontic lesions can be sterilized by the mixed drugs in situ. PMID- 8233572 TI - The in vitro lysozyme susceptibility of Candida albicans cultured in carbohydrate supplemented media. AB - The in vitro lysozyme susceptibility of three oral isolates of Candida albicans cultured in carbohydrate-supplemented media was studied. Lysozyme was shown to have a dose- and time-dependent killing effect on C. albicans isolates. Fungicidal activity persisted to varying degrees when yeast isolates were cultured in a variety of carbohydrates (glucose, galactose, sucrose, maltose, xylitol and lactose) before exposure to 20 micrograms/ml lysozyme. Sucrose and galactose grown yeasts exhibited increased resistance to lysozyme compared with (in decreasing order) those grown in glucose, maltose, xylitol or lactose. Further, the C. albicans isolates tested demonstrated strain variations in their susceptibility to lysozyme. These results suggest that dietary carbohydrate may play a role in modulating the yeast cell populations in the oral cavity by altering the fungal susceptibility to salivary lysozyme. PMID- 8233573 TI - Sensitization of periodontopathogenic bacteria to killing by light from a low power laser. AB - Cultures of Porphyromonas gingivalis, Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans were treated with a range of photosensitizers and then exposed to light from a 7.3 mW helium/neon laser for up to 80 s. Toluidine blue O (25 micrograms/ml) and methylene blue (25 micrograms/ml) were effective lethal photosensitizers of all 3 target organisms, enabling substantial light dose related reductions in viable counts. Dihaematoporphyrin ester and aluminium disulphonated phthalocyanine were lethal photosensitizers only of P. gingivalis. In the absence of a photosensitizer, exposure to laser light had no significant effect on the viability of the cultures. If such low doses of light (22 J/cm2) are effective at killing bacteria in vivo, the technique may be useful as a means of eliminating periodontopathogenic bacteria from diseased sites. PMID- 8233574 TI - In vitro adherence of rat lymphocytes to salivary gland epithelia. AB - Glandular mucosal tissues contain lymphocyte populations that contribute to expression of IgA antibodies in external secretions. Interaction of circulating lymphocytes with glandular structures may regulate lymphocyte accumulation. An in vitro assay was used to investigate adhesive interactions between lymphocytes and salivary gland tissues. Thoracic duct lymphocytes (TDL) bound to the serous acinar epithelia of parotid salivary glands and to the mucous tubulo-acinar epithelium of submandibular salivary glands. Lymph node cells and splenocytes adhered to these tissues in lesser numbers and thymocytes bound in negligible numbers. TDL adherence was an active process, being time- and cell dose-dependent and requiring intact membrane as well as cytoskeletal and metabolic function. Calcium was required in each case and binding was mediated by a trypsin-sensitive lymphocyte surface determinant. These findings suggest that the lymphocyte composition of salivary gland tissues is regulated by active lymphocyte interaction with the glandular epithelium. PMID- 8233575 TI - Nursing management of a child with juvenile rheumatoid arthritis. AB - Juvenile rheumatoid arthritis is an incurable chronic disease of childhood which involves connective tissue in the joints. Treatment aims are supportive and directed toward pain management, alleviation of inflammation, and optimization of joint function. A multidisciplinary team approach is helpful, with nursing care an essential part of the treatment plan. PMID- 8233576 TI - Subjective data related to altered bowel elimination patterns among hospitalized elder and middle-aged persons. AB - This exploratory study reports subjective data about bowel elimination pattern(s) (BEP) for elders admitted to an acute care hospital. This sample of a population from northern New England (n = 303) was collected from a small rural hospital and a large university hospital. A comparison group of 149 middle-aged subjects (40 to 64 years) was used to highlight responses of the elders (> or = 65 years). Data were collected through interviews and questionnaires and analyzed via content analysis. More than twice as many elders as middle-aged subjects reported a regular bowel habit as important to health. Significant changes in BEP were found to occur in both hospitalized elders and middle-aged subjects. Nurses need to pay careful attention to the BEP of these patients. Nurses should actively investigate BEP of subjects with increased risk and intervene with patient education or appropriate therapy as needed. PMID- 8233577 TI - Hypothermia and trauma. AB - Hypothermia can induce profound physiologic changes. There are varying degrees of hypothermia, each requiring aggressive methods of treatment. Trauma patients have multisystem involvement, and hypothermia can increase the morbidity and mortality of the trauma patient. The degree of hypothermia is affected by age, severity of illness, environmental exposure, and the presence of open wounds. It is important for all nurses who work with trauma patients in the perioperative phase to incorporate the assessment and planning to prevent or minimize hypothermia in the trauma patient. PMID- 8233578 TI - Program planning and evaluation: principles and procedures for nurse managers. AB - This article provides an overview of essential principles and procedures of program planning and evaluation. The relevance of these concepts to the task of health and nursing care is discussed in terms of three primary evaluation domains: (1) estimating need and designing evaluable programs; (2) determining the extent of program implementation; and (3) assessing program-related outcomes. The article has relevance for nurse managers concerned with programmatic decision making and accountability. PMID- 8233579 TI - Salmon calcitonin. AB - Calcitonin has a specific role in prevention and treatment of osteoporotic fractures. Calcitonin is a hormone that is synthesized and secreted by the thyroid gland. The circulating level of ionized calcium controls calcitonin release. Treatment of osteoporosis with calcitonin was approved by the FDA in 1985 (Fatourechi & Health, 1987). Calcitonin therapy suppresses bone loss and in some cases increases bone mass. Since intranasal and suppository routes are not approved in the United States, the primary route of administration is subcutaneous. In increasing incidence, nurses are teaching patients to self administer this medication. This article describes the purpose, action, and outcomes of calcitonin therapy and the required education needed by patients receiving this medication. PMID- 8233580 TI - Continuity of care: the orthopaedic nurse practitioner in tertiary care. AB - Since 1965 registered nurses have been educated to perform expanded functions in primary health care. The nurse provider role evolved from a shortage of primary health care providers. "Nurse practitioners" were needed to increase access to health care in urban and rural areas of the country that lacked the necessary number of providers to give quality patient care. Until recently, the majority of nurse practitioners have continued in the traditional primary care role, practicing in clinics, private physician offices, and on an independent basis. However, many factors now provide the impetus to have nurse practitioners develop tertiary care practices within the nation's acute care hospitals. This article describes the development and implementation of an orthopaedic collaborative practice in tertiary care. PMID- 8233581 TI - Vertical shear fractures of the pelvis. PMID- 8233582 TI - Letter from the management SIG post-congress 1993. PMID- 8233583 TI - MP implants for rheumatoid arthritis of the hand. AB - Rheumatoid arthritis is a devastating disease, which can incapacitate a person in activities of daily living. Only after all other methods have been exhausted, should patients undergo surgery for pain relief, mobility, and cosmetic reasons. A lengthy rehabilitative period following the placement of the metacarpophalangeal implants assures maximum mobility in the new joints. Nurses must be aware of their educational responsibilities to the patient before, during, and after the surgery for patient compliance. PMID- 8233584 TI - The mini-exon gene: a molecular marker for Endotrypanum schaudinni. AB - The mini-exon gene repeats from three different strains of Endotrypanum schaudinni have been amplified by the polymerase chain reaction (PCR). Sequence analysis of the cloned products shows the gene and intergenic region to be identical in two of the strains (LV86 and M6159); the intergenic region from the mini-exon gene of the third strain (LV59) is significantly different. The LV86 gene and an intergenic probe from the LV59 mini-exon gene do not cross-hybridize with the mini-exon gene from New World Leishmania species. These data provide the basis of a PCR assay to detect E. schaudinni and distinguish it from New World Leishmania species, and which should be applicable to epidemiological studies in insect vectors and mammalian reservoirs. The identification of two different mini exon gene repeats in E. schaudinni isolates is indicative of further strain variation within this species. PMID- 8233585 TI - The effect of transmission-blocking antibody ingested in primary and secondary bloodfeeds, upon the development of Plasmodium berghei in the mosquito vector. AB - The effects of purified monoclonal immunoglobulins from control, or transmission blocking anti-Pbs21 antibodies, upon the infection of Anopheles stephensi by ookinetes of Plasmodium berghei are compared. Anti-Pbs21 antibody reduced mean intensity and prevalence of infection by 94.7 and 58.7% respectively if added to the infectious bloodfeed at a concentration of 100 micrograms/ml. Fab fragments were of similar efficacy. No transmission enhancement was detected with declining antibody concentrations. Addition to subsequent (second) feeds reduced mean oocyst intensity but not prevalence. The reduction in blockade declined from 41% at day 2, to 4% at day 8. Second bloodfeeds, containing control globulin taken 4 or 6 days (but not 2 days) after infection, increased sporozoite burden in the salivary glands. At all times anti-Pbs21 reduced sporozoite number in the thorax compared to time-matched controls, but again highest gland intensities were obtained when the second bloodfeed was given on day 4. We conclude that second bloodfeeds containing transmission-blocking antibody simultaneously serve two opposing roles, (1) inhibition of parasite development and (2) the supply of nutrients which permit more sporozoites to be produced by each oocyst. PMID- 8233586 TI - Response of inbred mice to infection with a new isolate of Trypanosoma musculi. AB - A new isolate of Trypanosoma musculi has been derived from organisms recovered from Mus spretus captured in Lisbon, Portugal. The time-course and profiles of infection with this isolate in inbred mice have been compared with those obtained with the existing Partinico II isolate. Infections with the Portuguese isolate are less intense, and controlled more quickly than those with the Partinico isolate. As with the latter, there are marked mouse strain-dependent influences on infection with the Portuguese isolate, but these strain-dependent characteristics differ considerably with each isolate. For example, NIH mice were the most susceptible to the Partinico II isolate, but virtually refractory to the Portuguese isolate. Mice exposed to infection with one isolate show complete immunity to both homologous and heterologous challenge infections. These striking interactions between host and parasite genotype are discussed in terms of immunological influences on infection. PMID- 8233587 TI - Trypanosoma brucei brucei: differences in the nuclear chromatin of bloodstream forms and procyclic culture forms. AB - Nucleosome filaments of two stages of the life-cycle of Trypanosoma brucei brucei, namely bloodstream forms and procyclic culture forms, were investigated by electron microscopy. Chromatin of bloodstream forms showed a salt-dependent condensation. The level of condensation was higher than that shown by chromatin from procyclic culture forms, but 30 nm fibres as formed in rat liver chromatin preparations were not found. Analysis of histones provided new evidence for the existence of H1-like proteins, which comigrated in the region of the core histones in SDS-PAGE and in front of the core histones in Triton acid urea gels. Differences were found between the H1-like proteins of the two trypanosome stages as well as between the core histones in their amount, number of bands and banding pattern. It can be concluded that T. b. brucei contains a full set of histones, including H1-like proteins, and that the poor condensation of its chromatin is not due to the absence of H1, but most probably due to histone-DNA interaction being weak. It is obvious that structural and functional differences of the chromatin exist not only between T. b. brucei and higher eukaryotes, but also between various stages of the life-cycle of the parasite. It is therefore not adequate to investigate the chromatin only of the procyclic culture forms as a model for all stages of the life-cycle of T. b. brucei. PMID- 8233588 TI - Transmission of Theileria parva to cattle by Rhipicephalus appendiculatus adults fed as nymphae in vitro on infected blood through an artificial membrane. AB - A technique is described for the efficient feeding of Rhipicephalus appendiculatus nymphae on cattle blood through an artificial membrane bearing tactile and olfactory stimuli. The effect of four anticoagulation methods on the feeding of nymphae was compared and heparinized blood was found to be the most efficacious, followed by defibrinated blood. Blood treated with acid citrate dextrose (ACD) or ethylenediamine tetraacetate (EDTA) inhibited nymphal feeding. Nymphae fed on heparinized and defibrinated blood obtained engorgement weights within the range of ticks fed on mammalian hosts and they subsequently moulted and fed normally as adults and produced viable eggs. Nymphae fed on membranes using either defibrinated or heparinized blood infected with Theileria parva piroplasma developed salivary gland infections as adult ticks and transmitted East Coast fever (ECF) to susceptible cattle. There were indications that T. parva-infected defibrinated blood was not as infective to the feeding nymphae as the infected heparinized blood. When T. parva-infected heparinized blood was used to feed nymphae through membranes in two experiments, it was found that the infections in the resultant adult ticks could be comparable to those of nymphae fed on donor cattle, but were usually lower. The membrane feeding technique will enable the study of factors affecting the tick and T. parva transmission without the complication of host/T. parva interactions and could be useful for both tick maintenance and Theileria parasite isolation and maintenance. PMID- 8233589 TI - In vitro feeding of instars of the ixodid tick Amblyomma variegatum on skin membranes and its application to the transmission of Theileria mutans and Cowdria ruminatium. AB - An in vitro feeding method using rabbit or cattle skin membranes, applied successfully to all stages (larvae, nymphae and adults) of the ioxodid tick, Amblyomma variegatum, is described. The feeding apparatus consisted of a blood container with a membrane placed on top of a tick containment unit. A carbon dioxide atmosphere of between 5 and 10% and a temperature of 37 degrees C were used as stimulants for the attachment of the ticks. High CO2 concentrations in the atmosphere improved the feeding success of all instars. The effect of anticoagulation methods for the bloodmeal was investigated, and heparinized blood was found to be the most suitable for tick feeding. When the bloodmeal was replaced by tissue culture medium for feeding nymphs the subsequent moulting success was reduced. Adult ticks of both sexes remained attached for up to 16 days, until completion of their bloodmeals. All stages of the tick fed on whole blood in the artificial feeding system and all reached engorged weights less than those achieved by control ticks fed on experimental animals. A large proportion of ticks, fed artificially on whole blood, moulted or laid eggs successfully. The method was successfully applied for the transmission of Theileria mutans and Cowdria ruminantium to cattle. PMID- 8233590 TI - Protein synthesis and release by cultured schistosomula of Schistosoma mansoni. AB - The lung schistosomulum of Schistosoma mansoni is the target of protective immunity in mice singly vaccinated with irradiated cercariae. Since the effector responses are T cell-mediated, their initiation requires the release of antigens from the intact parasite. We have used the technique of biosynthetic labelling with [35S]methionine, before and after transformation of the cercariae, to analyse the kinetics of protein synthesis and release by the schistosomulum. In addition, the proteins present in the soluble fraction of the parasite and those released during in vitro culture have been characterized. During a 7-day culture period schistosomula derived from labelled cercariae lost proteins most rapidly within the first 3 h after transformation. Two proteins of molecular weight 61 and 20 kDa were dominant and may correspond to areas of proteolytic activity. Analysis of the rate of protein synthesis of schistosomula labelled after transformation revealed four different phases, which may relate to the developmental processes occurring in vivo. During the first 24 h, synthesis was very low, increasing to a plateau and then rising to a peak at day 8; therefore the rate declined rapidly. Whilst some stage-specific synthesis of proteins was detected in the soluble fractions of the parasite bodies, the pattern of proteins released by cultured larvae was remarkably uniform. At least 15 proteins were detected by autoradiography with bands at 61, 45 and 20 kDa being particularly prominent. These proteins merit further study as potential mediators of the protective immune response. PMID- 8233591 TI - Biochemical and morphological pathology of the foot of the schistosome vector Biomphalaria glabrata infected with Schistosoma mansoni. AB - Infection by Schistosoma mansoni resulted in morphological and biochemical changes to the foot of its intermediate host, Biomphalaria glabrata. Migration through, and emergence of cercariae from, the foot was observed and evidenced by lesions on the ciliated foot surface. This was accompanied by a significant decrease in the velocity of movement by infected individuals. In vivo 31P NMR spectral analyses demonstrated that the foot of infected snails had a lower phosphoarginine (PA)/adenosine 5' triphosphate (ATP) ratio than that of uninfected controls. Moreover, kinetic experiments, employing saturation transfer, demonstrated the pseudo-first-order rate constant for the arginine kinase-catalysed exchange reaction in the forward direction, that is, PA-->ATP was decreased by infection. The reverse reaction was not observed by the NMR methods used. PA was depleted upon exposure to hypoxic conditions suggesting its traditional role in preserving ATP level. Partly oxidized metabolic end-products were not observed in snails maintained under aerobic conditions, but succinate, propionate, acetate and lactate rapidly accumulated under hypoxic conditions. PMID- 8233592 TI - Biased sex ratios and parasite mating probabilities. PMID- 8233593 TI - Fasciola hepatica: the effect of the microtubule inhibitors colchicine and tubulozole-C on the ultrastructure of the adult fluke. AB - The effect of the microtubule inhibitors colchicine (1 x 10(-3) M) and tubulozole C(1 x 10(-6) M) on the ultrastructure of adult Fasciola hepatica has been determined in vitro by transmission electron microscopy (TEM), using both intact flukes and tissue-slice material. With colchicine treatment, the apical membrane of the tegument became increasingly convoluted and blebbed, while accumulations of T1 secretory bodies occurred in the basal region of the syncytium, leading to progressively fewer secretory bodies in the syncytium. In the tegumental cells there were distinct accumulations of T1 secretory bodies around the Golgi complexes, which remained active for up to 12 h incubation. Tubulozole-treated flukes showed more severe effects, with initial accumulations of secretory bodies, both at the tegumental apex and base. This was followed in the later time periods by the sloughing of the tegumental syncytium. In the underlying tegumental cells, the granular endoplasmic reticulum (GER) cisternae were swollen and disrupted, becoming concentrated around the nucleus. The Golgi complexes were dispersed to the periphery of the cells and gradually disappeared from the cytoplasm. After treatment with both drugs, the cell population in the vitelline follicles was altered, with an abnormally large proportion of stem cells and relatively few intermediate type 1 cells. The nurse cell cytoplasm became fragmented and was no longer in contact with the vitelline cells, while the shell globule clusters within the intermediate type 2 and mature cells were loosely packed. In the mature vitelline cells, 'yolk' globules and glycogen deposits became fewer than normal and lipid droplets were observed. The results are discussed in relation to the different modes of action of the two drugs and potential significance of this to anthelmintic (benzimidazole) therapy. PMID- 8233594 TI - Vaccination against Trichinella spiralis in mice using antigens from different isolates. AB - Antigen preparations from three isolates of Trichinella spiralis (sensu-stricto) have been tested for their ability to immunize mice against homologous and heterologous challenge infections. Immunization was measured by accelerated expulsion of adult worms, reduction in female worm fecundity, stunting of growth, and level of antibody responses before and after challenge. Antigens of two isolates, from London (L) and Poland (P), immunized effectively against homologous challenge, adult worm recoveries being reduced by 64% and 51% respectively within 6 days. Antigen from a Spanish (S) isolate gave no homologous protection in terms of worm recovery at day 6 and only 43% reduction at day 8. L- and P-isolate antigens immunized well against heterologous challenge (day 6 worm recoveries reduced by 66% and 59%) but similar protection against S-isolate challenge was not seen until day 8. Immunization with S antigen gave better protection at day 6 against L-isolate challenge (77%) than against challenge with P-isolate (37%). These results imply considerable, but not complete, antigen cross-reactivity between the isolates and this was reflected in antibody responses. These variations in immunogenicity and immunization potential are discussed in terms of constraints on the use of vaccines against parasites that are widely distributed geographically. PMID- 8233595 TI - Immunological relationships during primary infection with Heligmosomoides polygyrus. Regulation of fast response phenotype by H-2 and non-H-2 genes. AB - The inheritance of response phenotype to Heligmosomoides polygyrus was investigated in F1 hybrid progeny of fast and slow responder mouse strains. The fast responses of SJL(H-2s) and SWR(H-2q) mice were mediated by dominant genes complementing each other in F1 hybrids which lost worms earlier and produced faster parasite-specific IgG1 antibody responses than either parent. However, the response of F1 hybrids from crosses of C57BL/10 (B10, H-2b) mice with either SJL or SWR differed from that of the parental strains and from each other: (B10 x SWR)F1 lost worms earlier than SWR whilst (B10 x SJL)F1 lost worms later than SJL mice. The F1 progeny of SJL mice with congenic strains B10.G (H-2q) and B10.S (H 2s) lost worms as quickly as SJL. Therefore, the response phenotype mediated by the genome of SJL mice was unaffected by H-2 heterozygosity (with H-2q) or homozygosity (H-2s) despite heterozygosity with B10 background genes, but was slowed significantly by heterozygosity with H-2b. All hybrids involving heterozygosity with B10, irrespective of MHC haplotype or background, failed to clear worms completely, in each case a proportion of mice harbouring residual worm burdens after loss of worms from parental strains. PMID- 8233596 TI - Commission of the European Communities 2nd Malaria Contract Holders Meeting. Orvieto, 20-22 March 1992. Proceedings. PMID- 8233597 TI - The epidemiology of malaria in southern Sierra Leone. AB - The epidemiology of malaria was investigated in a high rainfall, forested area of southern Sierra Leone. The prevalence rates of P. falciparum, P. malariae and P. ovale in 0-7 year old children, during two surveys conducted over a 12-month period, averaged 61%, 12% and 1% respectively. Groups of febrile children had higher prevalence rates than afebrile groups. Overall, gametocyte rates were approximately one fifth of the trophozoite rates. Malaria accounted for 27% of deaths, as did malnutrition, although no malaria associated deaths occurred in 0 12 month olds. Spleen rates were similar to P. falciparum prevalence rates, and the size did not appear to be related to parasite load at the time of the surveys. Packed cell volumes had normal distributions, with a lower mode after the peak prevalence period. Chloroquine usage increased during the post-rains period compared to the pre-rains period. PMID- 8233598 TI - Plasmodium falciparum proteinases and red blood cell invasion. AB - Malarial proteinases of the erythrocytic life-cycle are used to design new inhibitors capable of blocking the parasite's development. The Merozoite Proteinase for Erythrocytic Invasion (MPEI) of Plasmodium falciparum, a neutral proteinase, and the acidic Pf37 proteinase acting on spectrin as substrate, are good candidates for this kind of strategy. PMID- 8233599 TI - Population dynamics of human malaria parasites. AB - Highly sensitive detection and identification of the four Plasmodium species infecting man, and the characterization of P. falciparum isolates, have been achieved by PCR amplification. The results obtained from field and laboratory studies are described. The significance of these observations to the design and interpretation of epidemiological investigations is discussed. PMID- 8233600 TI - Sensitive detection of Plasmodium falciparum in blood and mosquito by DNA amplification. AB - By means of enzymatic amplification of Plasmodium falciparum DNA using the polymerase chain reaction (PCR), we have been able to detect as little as 20 parasites in 20 ml infected human blood based on the visualization of a 206 bp fragment in ethidium bromide-stained agarose gel. Comparison, through microscopic examination of 2030 blood samples collected from various endemic areas in Thailand, indicates that the PCR-based detection system is 96% specific and 81% sensitive, with a disagreement of 6.6%. The same protocol can be applied to detect a minimum of 10 sporozoites or a single oocyst dissected from P. falciparum-infected mosquito. PMID- 8233601 TI - Phosphorylation of infected erythrocyte membrane proteins in Plasmodium chabaudi. AB - We identify phosphoproteins that are associated to the infected erythrocyte membrane of Plasmodium chabaudi. The specific antibodies that recognize the erythrocyte membrane-associated antigens, namely PcEMA1 and PcLEMA, are studied. According to sequencing analysis, neither the gene structure nor the predicted properties of these two antigens are completely distinct. The PcEMA1 is a 50 kDa acidic protein, with a pI of about 4.4, that contains 25 phosphorylation sites principally located in the repetitive sequence. Almost all of this molecule is hydrophilic. The predicted (predictment ability) of this protein is, hence, associated to the host's erythrocyte cytoskeleton and it is synthesized through an entire erythrocyte cycle. PcLEMA, instead, is a 74 kDa basic protein, with a pI of about 9.8, that contains 11 possible phosphorylation sites. This gene contains two exons: exon 1 shows transmembrane characteristics while exon 2 contains part of the hydrophilic repetitive sequences. Thus this protein is predicted as a transmembrane-associated protein and is synthesized only at the last stages of the erythrocytic cycle. In in-vitro phosphorylation experiments, both PcEMA1 and PcLEMA are phosphorylated by endogenous kinases activities. However, the degree of phosphorylation differs between the two in that PcEMA1 reveals a higher phosphorylation intensity than PcLEMA. PMID- 8233602 TI - The design of original antimalarial drugs. An example of phospholipid metabolism. AB - The aim of our program was to find an original chemotherapeutical treatment (and eventually a preventive treatment) of malaria, an illness largely predominant in developing countries, by interfering on an essential metabolism developed by Plasmodium during its erythrocytic phase. Apart from what has been learnt about metabolism and the pharmacological target, a crucial step has been taken during this contract by passing from micromolar in vitro active concentrations (during 1986-1990) to nanomolar ones (during 1990). These compounds should naturally short-circuit resistance phenomena already established against drugs in current use, as has already been verified on polypharmacoresistant strains or isolates of P. falciparum. The administration of a therapeutic dose of our molecules would now appear to be possible in all cases. PMID- 8233603 TI - Antigenic diversity, antigenic variation and merozoite surface protein 1. AB - Antigenic diversity and immune evasion by the parasite are important factors in the epidemiology of malaria and in determining the incidence of severe malaria. The dynamics of the parasitaemia in current studies in mice immunized with P.c.chabaudi native MSP1 or recombinant C-terminal subunits thereof, strongly indicate evasion of components of the immune response induced by these antigens by parasites multiplying after challenge infection. PMID- 8233604 TI - Plasmodium vivax malaria in Sri Lanka: transmission, disease and immunity. AB - We have established a study site in a malaria endemic region around Kataragama in the south of Sri Lanka. A population of about 3,000 individuals in a group of villages around Kataragama have been provided with access to a small field clinic enabling us to monitor aspects of malarial infection under conditions of natural transmission. At the present time, both Plasmodium falciparum and Plasmodium vivax are prevalent to a similar degree although prior to about five years ago P. falciparum had been virtually absent for most of the previous ten years. PMID- 8233605 TI - Genetic study of the susceptibility of Anopheles gambiae to infection with malaria parasites. PMID- 8233606 TI - Advances in the study of Afrotropical malaria vectors. AB - The advances in the study of Afrotropical malaria vectors outlined in this report were obtained with the support of the EC-STD2 program by a network led by a research team of the Istituto di Parassitologia, University of Rome "La Sapienza" and including various collaborators based in different African countries. A list of authors and collaborating laboratories is given in the acknowledgements. The main results obtained are as follows. 1) Discovery of new chromosomally recognized taxa of the An. gambiae complex in Mali. One of them, the Mopti chromosomal form, is specifically adapted to breeding during the dry season and replaces other members of the complex in irrigated areas also in Burkina Faso and Northern Benin. 2) Evidence for increasing potential for P. falciparum transmission in the area of Cotonou (Benin) where highly anthropophilic An. gambiae (fresh water breeder) replaces the less effective vector An. melas (salt water breeder) when lagoon pile-dwelling traditional villages are converted into unplanned urban settlements. 3) Analysis of the vector system responsible for the P. falciparum epidemics on the Madagascar plateau showing the absence of An. gambiae, the secondary role of An. arabiensis (characterized by zoophilic and exophilic behaviour) and the primary role of An. funestus which is characterized by a peculiar focal distribution presumably depending on larval overwintering. 4) Evidence for a role for An. pharoensis (or at least of one of the taxa of this complex) in the transmission of P. falciparum malaria in the Senegal river delta. 5) Evidence for bionomical differences between An. gambiae and An. arabiensis and for intraspecific heterogeneities in both taxa associated with paracentric inversion polymorphisms. PMID- 8233607 TI - DNA probes for the identification of malaria vectors. AB - The correct identification of the mosquito vectors of malaria is essential to determine the vectorial significance of the different species within species complexes and to optimise vector control programmes. A number of methods are available including cytogenetic analysis, isoenzyme characterisation, and mating incompatibility. We describe here the development of a simplified DNA probe methodology for the identification of malaria vector specimens, with particular emphasis on the identification of mosquitoes of the Anopheles gambiae complex, which includes the most important vectors of malaria in Africa. The method incorporates the use of a simple 'squash-blot' sample preparation, synthetic oligonucleotide probes, non-radioactive labelling and detection systems, cheap and stable reagents, and all processes are carried out at room temperature without the need for equipment. Identification of up to 300 specimens (larvae, pupae or adults) per day is possible by a single worker using this simple technique and costs $0.02-$0.04 per specimen. PMID- 8233608 TI - Lymphocyte response in vitro to Plasmodium falciparum merozoite antigens in donors from a holoendemic area. AB - Crude merozoite antigens from P. falciparum were used to analyse the proliferative response of peripheral blood mononuclear cells from 114 inhabitants of the village of Dielmo (Senegal, West Africa), who are exposed continuously to malaria transmission. The high or low responses to merozoite antigens obtained in lymphocyte stimulation assays were correlated to the presence or absence of parasites, to the IFN-gamma production and to the HLA-phenotype. High responders produced high levels of IFN-gamma while low responders did not secrete IFN-gamma (23/27). The two HLA phenotypes HLA-B51 and HLA-DR1 were significantly associated with high response (p < 0.05). PMID- 8233609 TI - Mobile repeat units in Plasmodium berghei. PMID- 8233610 TI - The role of the host during the development of Plasmodium berghei hepatic schizonts. AB - Immature exoerythrocytic stages of Plasmodium berghei are immunogenic and produce antigens with protective capacities. Immunization experiments show a strong dependency of the responses on the host species and strain. To study this dependency a potential natural host of Plasmodium berghei, Thamnomys gazellae, was introduced, a species which is very susceptible for infection. Young liver stages were produced in different hosts after inoculation with irradiated sporozoites or after treatment with difluoromethylornithine. PMID- 8233611 TI - Glutamate rich Plasmodium falciparum antigen (GLURP). AB - We have characterized a glutamate rich Plasmodium falciparum antigen, GLURP, which by immunoassays appears to be present in both the pre-erythrocytic and erythrocytic stages of the vertebrate life cycle. The gene, which is located on chromosome 10, encodes a polypeptide of 1271 residues with a predicted molecular mass of 145 kDa. Rabbit antiserum against a fusion protein expressing the C terminal end of the molecule detects a protein with a molecular mass of 220 kDa. The sequence includes two hydrophobic regions: one consisting of 23 residues and located by the N-terminus which may act as signal peptide, and the second located at the C-terminus consisting of 33 predominantly hydrophobic residues. Except for these hydrophobic regions the protein is hydrophilic and highly charged. The sequence has two tandem repeats designated as R1 and R2. These regions were found to be conserved in isolates from different geographical areas. High levels of anti-GLURP antibodies have been shown to correlate with low parasite density. The indication of GLURP being present in all stages of the parasite in the human host raises significantly the prospects of the potential of this molecule. PMID- 8233612 TI - The ecology and behaviour of the forest form of Anopheles gambiae s.s. AB - Studies on the ecology of Anopheles gambiae s.s. and the transmission of malaria were undertaken in a high rainfall forested area in southern Sierra Leone. Anopheles gambiae s.s., identified by chromosomal techniques as the Forest form, was the only malaria vector in the study village. Surprisingly, rice fields or swamps were not favoured breeding places for this species; breeding mainly occurred in temporary pools. The mean annual sporozoite rate of An. gambiae s.s. determined by ELISA was 3.9%. Pyrethrum spray, human bait, and exit trap collections, as well as identification of mosquito blood-meals using the ELISA method, showed that the forest chromosomal form of An. gambiae s.s. was highly anthropophagic and exophilic. PMID- 8233613 TI - Genome organization, chromosome translocation and size polymorphism in rodent malaria parasites. AB - In our laboratory, rodent malaria models are used to investigate processes that underlie cell differentiation with specific emphasis on sexual development. The rodent parasite Plasmodium berghei is particularly suited for this research since the different sexual stages (from young gametocytes to mature ookinetes) can be obtained pure and in large numbers. PMID- 8233614 TI - Chronobiology and chronotherapy of malaria: investigations with murine malaria models. AB - The schizogonic rhythm of rodent malarias in the blood of mice varies from one species or subspecies to the other. Synchronous strains enable a precise study of the sensitive stage of the parasite to antimalarial drugs. Asynchronous strains are less sensitive than synchronous strains because of the delayed penetration of merozoites into red blood cells. PMID- 8233615 TI - Hepatic stages of malaria: specific and non-specific factors inhibiting the development. AB - Protection against pre-erythrocytic stages of malaria is possible, as demonstrated by the resistance obtained by immunizing with irradiated sporozoites. However, the involved mechanisms are more numerous and intricate than previously believed. Recently, the hepatic stage, rather than the sporozoite stage, has been seen as the target of immune attack. PMID- 8233616 TI - Malaria transmission-blocking activity in the plasma of Plasmodium falciparum gametocyte carriers in Cameroon. AB - Experimental infections of Anopheles gambiae were carried out with Plasmodium falciparum gametocytes from 65 naturally infected patients in Cameroon. A comparison was made between infections with blood containing autologous plasma and blood in which the plasma was replaced by plasma from donors without previous malaria exposure. A lower mosquito-infection rate was observed in 50 out of 65 autologous plasma samples. The transmission was completely blocked in 8 infections, whilst belonging exposures to heterologous plasma led to infected mosquitoes. Evidence is shown that blood plasma factors of gametocyte carriers from a population living in a malaria-endemic area are able to reduce transmission capacity. PMID- 8233617 TI - Thrombospondin related anonymous protein (TRAP) of Plasmodium falciparum in parasite-host cell interactions. AB - Thrombospondin related anonymous protein (TRAP) of Plasmodium falciparum is characterized by the presence of an amino acid motif based on the sequence Trp Ser-Pro-Cys-Ser-Val-Thr-Cys-Gly (WSPCSVTCG) that is found in a growing family of proteins. The sequence WSPCSVTCG is considered to confer sulpho-galactosyl cerebroside (sulphatide) binding properties to antistasin, TSP, CS protein and properdin. The observation that TRAP is localized both on the micronemes and on the surface of P. falciparum sporozoites would suggest a role played by TRAP, and its putative sulphated glycoconjugates binding motif, in the recognition and/or entry of hepatocytes by the sporozoite. Our results indicated that TRAP constructs, expressed in E. coli, bind to sulpho-galactosyl-cerebrosides (sulphatides) and to the surface of HepG2 cells using the conserved amino acid motif WSPCSVTCG. Antisera raised against TRAP constructs inhibited sporozoite invasion of HepG2 cells thus suggesting, thus, that TRAP may be one of the parasite-encoded molecules implicated in the sporozoite invasion of hepatocytes. Moreover, the possibility that TRAP antibodies may be relevant in malaria immunity is supported by the results obtained in a prospective study conducted in a malaria endemic area. In adolescents, the presence of TRAP antibodies, before malaria transmission, correlated positively with the control of parasite density. PMID- 8233618 TI - Temporal and spatial expression of serine protease genes in Anopheles gambiae. AB - Serine proteases play a crucial role during the digestion of the blood meal in the mosquito gut. The isolation and the analysis of the genomic organisation of the corresponding genes may lead to the characterization of gut-specific, inducible promoters, suitable for the expression of anti-parasitic agents in the gut of transgenic mosquitoes. We report here on the identification of a trypsin and a chymotrypsin gene family of Anopheles gambiae. Following a blood meal, the transcription of all members of the two identified gene clusters, seven trypsin genes (Antryp1-7) and two chymotrypsin genes (Anchym1-2), is induced. Recombinant Antryp1 and Antryp2, expressed in E. coli, were both active in vitro against blood proteins. Moreover, mouse sera raised against Antryp1, Anchym1 and Anchym2 recognized the corresponding proteases among the proteins of a lysate prepared from dissected guts of An. gambiae mosquitoes. PMID- 8233619 TI - Malaria sporozoites and circumsporozoite protein bind sulfated glycans: carbohydrate binding properties predicted from sequence homologies with other lectins. AB - Circumsporozoite (CS) proteins, the major surface proteins of the sporozoites of the various malaria (Plasmodium) species, share a region of highly conserved sequence homology in common with sporozoite surface protein 2 (SSP2) and a group of proteins observed to specifically bind sulfated glycoconjugates. Recombinant P. yoelii CS protein was found to bind selectively to heparin-, fucoidan-, and dextran sulfate-Sepharose, but poorly to chondroitin sulfate A- or C-Sepharose. It also bound with lower affinity to a heparan sulfate biosynthesis-deficient mutant cell line compared with the wild-type. Likewise, P. berghei sporozoite invasion into hepatocytes was selectively inhibited by fucoidan, heparin, and dextran sulfate, and sporozoites bound specifically to sulfatide [galactosyl (3 SO4) beta 1-1 ceramide] coated surfaces. Sporozoite infectivity in mice was significantly inhibited by dextran sulfate 500,000 and fucoidan. Taken together, these data indicate that CS proteins bind selectively to certain sulfated glyconjugates and invasion of host hepatocytes by sporozoites, and sporozoite infectivity can be inhibited by such compounds. PMID- 8233620 TI - Controlled trial of lambda-cyhalothrin impregnated bed nets and Maloprim chemosuppression to control malaria in children living in a holoendemic area of Sierra Leone, west Africa. Study design and preliminary results. AB - The effect of lambda-cyhalothrin impregnated bed nets and maloprim/placebo was studied in approximately 1,500 children living in 17 villages in a rural area of Sierra Leone, approximately 150 miles south east of Freetown, 30 miles north of the town of Bo. Villages were selected randomly amongst villages with impregnated bed nets and villages with no nets at all. Within these villages, children with ages ranging between 3 months to 6 years were chosen to receive maloprim or a double-blind distributed placebo fortnightly. In the villages randomised to receive nets, all beds have received nets. Malaria morbidity is estimated from weekly active case detection, and the impact on the Anopheles vector is being estimated by indoor spray catching, exit trap catching, human night landing catches and light trap catches. During the first 8 weeks of the intervention there was a significant reduction in slide positive rates, reported fever rates and children with temperature > or = 37.5 degrees C in the villages with impregnated bed nets. PMID- 8233621 TI - Chromosomal polymorphism and sexual differentiation in Plasmodium. AB - The correlation observed in several instances between the loss of ability to produce gametocytes and chromosomal rearrangements, prompted us to investigate in further detail the molecular bases of chromosomal polymorphism in Plasmodium. Generation of polymorphic karyotypes in Plasmodium involves important rearrangements, mostly occurring in subtelomeric position. Detailed analysis on the organisation of these regions have been carried out on the rodent malaria P. berghei and the human malaria P. falciparum. A 2.3kb sequence, tandemly organised in long clusters is shared by many P. berghei chromosomal ends. Variations in the copy number of this "module" account for most of the observed polymorphisms. In a P. falciparum cloned line (3D7) a common region spanning at least 40 kb, is present. It does not contain any repetitive structure other than the rep20 cluster, that appears to be completely contained within the common region. Notwithstanding the structural differences, human and rodent Plasmodia share the common feature of possessing long subtelomeric regions showing, thus, a homology between the different chromosomes. PMID- 8233622 TI - A study of the consequences of malarial infection in pregnant women and their infants. AB - An outline is given of a field research study to be undertaken in Malawi to investigate the pattern and consequences of malaria in pregnancy and infants. The central question to be investigated is whether babies born to anaemic mothers in malarious areas are at increased risk of developing anaemia or altered risk for morbidity from malaria or develop anaemia in the first year of life. The framework for the case control and cohort study to be undertaken is outlined. PMID- 8233623 TI - Polytene chromosome microdissection and molecular genome mapping in Drosophila and other dipterans. AB - We are constructing a molecular physical map of the Drosophila melanogaster genome, using microdissection of polytene chromosomes as a source of region specific probes applied to direct mapping work. Chromosome microdissection has been used for a low resolution map of the Anopheles gambiae s.s. genome. PMID- 8233624 TI - [Multicenter study of the in vitro sensitivity of genital mycoplasmas to antibiotics]. AB - The in vitro susceptibility of Ureaplasma urealyticum (Uu) and Mycoplasma hominis (Mh) was evaluated in a multicentric study performed in seven hospitals from different geographic areas in France. During a three month period, 324 Uu and 72 Mh clinical isolates were tested using a system ready for use, SIR Mycoplasma (Sanofi Diagnostics Pasteur). For Uu, the percentage of strains intermediate (I) or resistant (R) was as follows: doxycycline (3), minocycline (2.5), lymecycline (6.7), erythromycin (72, most I), josamycin (0.9), clindamycin (88), pristinamycin (0.3), ofloxacin (34, most I). For Mh, the percentage of strains I or R was respectively: doxycycline (2.7), minocycline (5.5), lymecycline (15.2), erythromycin (100), clindamycin (1.4), ofloxacin (2.7), josamycin (0) and pristinamycin (0). Comparable results were observed in the different geographic areas. The frequency of acquired resistances does not justify modifications in the usual treatment of genital mycoplasma infections but leads to monitor their susceptibility to antibiotics. PMID- 8233625 TI - [Compared antibacterial activity of a new fluoroquinolone, sparfloxacin (AT 4140, RP 64206) and four other fluoroquinolones against 332 strains of enteropathogen bacteria]. AB - The in vitro bacteriostatic and bactericidal activity of five fluoroquinolones- sparfloxacin, ciprofloxacin, ofloxacin, lomefloxacin and pefloxacin--was tested against 332 strains of enteric pathogens belonging to the genera Salmonella, Shigella, Escherichia, Yersinia, Vibrio, Campylobacter and Helicobacter. Some of the strains were resistant to one or several antibiotics. Each fluoroquinolone showed identical bacteriostatic activity against susceptible and resistant strains except of those resistant to nalidixic acid. MIC 90% were always below 1 mg/l. The MIC were lowest with ciprofloxacin followed by sparfloxacin and highest with pefloxacin. The five compounds showed different activity against nalidixic acid resistant strains: MIC 90% increased from 0.06 to 4 mg/l for Salmonella, from 0.5 to 4 mg/l for C. jejuni strains, from 0.12 to 16 mg/l for H. pylori strains. All strains remained susceptible to ciprofloxacin and sparfloxacin, but some were intermediate or resistant to the three other compounds. The minimal bactericidal concentrations of the five agents against the nalidixic acid susceptible or resistant strains were one or two times the corresponding MIC. PMID- 8233626 TI - [Action of teicoplanin on coagulase negative staphylococci in hospital units of the hospital Hotel-Dieu in Paris]. AB - Teicoplanin, a glycopeptide antibiotic, is active against Staphylococci, with the exception of some strains of coagulase negative Staphylococci (SCN). Determination of the in vitro activity of teicoplanin by standard disk methodology is not sufficient, and has to be confirmed by the determination of the minimal inhibitory concentration (MIC) for all strains presenting an inhibition zone diameter below 17 mm. We have studied during four months, the susceptibility of SCN to teicoplanin isolated from different units of the hospital, and in particular from the haematology unit (HU). 186 strains were isolated, 35 p. cent from HU and 65 p. cent from the other units of the hospital. All strains belonged to 11 species: 136 S. epidermidis, 14 S. haemolyticus, 8 S. lugdunensis, 5 S. warneri, 4 S. saprophyticus, 3 S. sciuri, 3 hominis, 2 S. chromogens, 2 xylosus, 2 S. cohnii and 1 S. schleiferi. Twenty five strains out of 186 (13.4%) presented a MIC of 8 or 16 mg/l (MIC verified by the agar dilution method): 7 S. haemolyticus, 16 S. epidermidis, 1 S. warneri et 1 S. capitis. During this study SCN resistant to teicoplanin were never isolated. Twelve out of 25 came from the HU, which correspond to 18 p. cent of the isolated strains in this unit. This result was not significantly different, compared to that of the other units. PMID- 8233627 TI - [In vitro study of the sensitivity to antibiotics of 213 Pseudomonas aeruginosa strains isolated in an intensive care unit of the Sfax Hospital (Tunisia)]. AB - Susceptibility of 213 strains of Pseudomonas aeruginosa isolated in a general intensive care unit during two years 1989 and 1990 was tested against 11 antibiotics: ticarcillin (TIC), ticarcillin+clavulanic acid (TCC), cefsulodin (CFS), ceftazidime (CAZ), imipenem (IMP), gentamicin (G), tobramycin (TOB), amikacin (AN), ofloxacin (OFX), norfloxacin (NOR) and ciprofloxacin (CIP). Antibiotic susceptibility testing was performed by disk agar diffusion test and by measuring minimum inhibitory concentrations (MICS using test agar dilution. 65% of strains were isolated from respiratory tract infections. Resistance frequency of this antibiotics was: TIC: 41.3%, TCC: 33.8%, CFS: 26.5%, CAZ: 23.2%, IMP: 11.6%, G: 72.5%, TOB: 46.5%, AN: 12.6%, OFX: 29.3%, NOR: 21.3%, CIP: 17.6%. We observed by measuring CMI that imipenem and amikacin were the most active antibiotics. From 1989 et 1990, the increase number of strains resistant to imipenem, amikacin and fluoroquinolones could be explained by massive use of those antibiotics. Besides their high level of resistance, strains isolated in intensive care unit ere characterized by their multiresistance: 24 strains were resistant to 8 of 11 antibiotics tested; four of them showed resistance to all antibiotics the multiresistance of those strains suggest strongly that decreased permeability could be the cause. PMID- 8233628 TI - In vitro evaluation of activities of azithromycin, clarithromycin and sparfloxacin against Chlamydia trachomatis. AB - The in vitro activities of azithromycin, clarithromycin and sparfloxacin were evaluated by studying inhibition of in vitro Chlamydia trachomatis propagation in McCoy cells, comparatively with erythromycin, ofloxacin and tetracycline. Fifteen clinical isolates of C. trachomatis were tested with an inoculum of 5.10(3) inclusion--forming units in a 96--well microtiter plate. Minimal inhibitory concentration (MIC) ranges were as follows: azithromycin, 0.06 to 0.125 microgram/ml; clarithromycin, 0.008 microgram/ml; erythromycin, 0.06 to 0.125 microgram/ml; ofloxacin 0.5 to 1 microgram/mg; sparfloxacin, 0.03 to 0.06 microgram/ml; and tetracycline 0.125 to 0.25 microgram/ml. Minimal bactericidal concentration (MBC) ranges, calculated from passage into antibiotic--free medium, were as follows: azithromycin 0.25 to 0.5 microgram/ml; clarithromycin, 0.03 to 0.125 microgram/ml; erythromycin, 0.25 to 2 micrograms/ml; ofloxacin, 0.5 to 1 microgram ml; sparfloxacin, 0.03-0.06 microgram/ml; and tetracycline, 1 to 4 micrograms/ml. Clarithromycin and sparfloxacin showed the greatest activity and clinical studies of these agents in C. trachomatis infections are therefore indicated. PMID- 8233629 TI - [Ciprofloxacin, clarithromycin, rifampin: in vitro bactericidal activity of double and triple drug combinations against strains of atypical mycobacteria]. AB - Bactericidal activity of ciprofloxacin (CIP), clarithromycin (CLA), rifampin (RIF) alone and in combinations was studied against five atypical mycobacteria (4 isolates from patients with Acquired Immunodeficiency Syndrome and 1 collection strain) by time killing curves method. Drugs were used at their attainable serum levels: CIP (1 microgram/ml), CLA (2 micrograms/ml), RIF (16 micrograms/ml). The decrease in CFU in comparison to inoculum (10(6)-10(7) CFU/ml) was evaluated by viable counts after 1, 2, 3, 4 and 5 days of incubation at 35-37 degrees C. Drug combinations enhanced slightly the killing of drugs alone. CIP+CLA was the most effective against MAC strains whereas CIP+RIF appeared as the best combination against rapidly growing species except M. phlei which was the most resistant strain. PMID- 8233630 TI - Determination of MICs for staphylococci using the API ATB quinolone and API ATB macrolide systems. AB - The determination of minimal inhibitory concentrations (MICs) is cumbersome, but remains necessary in certain cases. We tested the two ATB MIC experimental strips (Biomerieux SA), of which each contains 4 antimicrobials of the same class. These strips can be read automatically. The MIC quinolone strip contains nalidixic acid, pefloxacin, ofloxacin and ciprofloxacin, whereas the MIC macrolide (lincosamide-streptogramin) strip contains erythromycin, clindamycin, lincomycin, and pristinamycin. In order to evaluate these strips, 102 S. aureus and 63 coagulase negative staphylococci were used. Correlation coefficients for these MICs (micrograms/ml) and disk diffusion inhibition zone diameters (mm) were nalidixic acid -0.59, pefloxacin -0.95, ofloxacin -0.95, ciprofloxacin -0.91, erythromycin -0.98, clindamycin -0.96, lincomycin -0.96, and pristinamycin -0.64. Using the Biomic system (Giles Scientific USA), the same zone diameters were converted to MICs (micrograms/ml). Rates of agreement (+/- 1 dilution) between ATB MICs and Biomic MICs were nalidixic acid 96 p. cent, ciprofloxacin 98 p. cent, erythromycin 99 p. cent and clindamycin 98 p. cent. Rates of agreement between MICs for the same strains determined using agar dilution and ATB MICs were nalidixic acid 93 p. cent, pefloxacin 100 p. cent, ciprofloxacin 99 p. cent, ofloxacin 94 p. cent and erythromycin 96 p. cent. The ATB MIC strips are an easy to-use tool for MIC determination and their composition is well-suited to the study of phenotypic resistance and detection of low-level resistance. PMID- 8233631 TI - [Detection of the phenotypes of resistance of enterobacteriaceae to aminoglycosides with ATB Plus Expert System]. AB - ATB Plus Expert (Biomerieux SA) is an expert system which has been developed to perform an interpretative reading of ATB susceptibility tests. The system was tested on the results obtained for 217 strains of enterobacteriaceae. These strains were selected in order to cover a maximum of bacterial species and resistance mechanisms. The isolates were tested on rapid ATB E, rapid ATB G-, rapid ATB Ur, ATB G- and ATB Ur strips. In parallel, a disc diffusion susceptibility test was performed with 5 discs of aminoglycosides (kanamycin, gentamicin, tobramycin, netilmicin, amikacin) and the interpretation was carried out according to the criteria usually followed. Of the 217 strains tested, 122 showed a resistance phenotype. Only the rapid ATB E strips included kanamycin and allowed the detection of APH(3') phenotypes. Amikacin was not included in the ATB Ur strip, consequently it was impossible to discriminate AAC(3)-II and AAC(6') + AAC(3)-I phenotypes. 12 strains did not grow within 5 hours using the rapid ATB methodology. Not taking into account the problems previously encountered, different phenotypes between the 6 susceptibility tests were found for 16 strains. In 5 cases the expert system detected an anomaly instead of the correct phenotype, and in 3 cases of unknown phenotypes, the answers were variable. In the other cases, the main difficulty was the detection of the isolated resistance to gentamicin (AAC(3)-I phenotype). The expert system automatically corrects the susceptibility test result according to the phenotype observed. PMID- 8233632 TI - [Multifactorial analysis of the phenotypes for beta-lactams of 1044 Escherichia coli strains]. AB - 1,044 E. coli strains were randomly collected by the beginning of 1992. Their susceptibility for seven beta-lactam antibiotics: amoxycillin, augmentin, ticarcillin, claventin, cephalothin, cefoxitin and cefotaxime, was studied routinely by the agar diffusion method. The datas were analyzed by the CERIB multifactorial analysis package which yields to homogeneous populations. This analysis showed four well defined populations: 1) 588 strains (56.4%) susceptible to all antibiotics; 2) 410 strains (39.3%) present a penicillinase phenotype; 3) 11 strains (1.05%) are cephalosporinase producer; 4) 7 strains (0.67%) were identified as producing an extended-spectrum beta-lactamase. The remaining strains: 28 (2.68%) had a reduced susceptibility to all antibiotics, which suggests the combination of few resistance mechanisms or other hypothesis. PMID- 8233633 TI - [Determination of the isoelectric point of beta-lactamases isolated from 67 Klebsiella oxytoca strains and phenotype behaviour against eight beta-lactam antibiotics]. AB - Klebsiella oxytoca is naturally resistant to aminopenicillins and carboxypenicillins by production of a chromosomal beta-lactamase but susceptible to third generation cephalosporins. The third generation cephalosporins activity may be reduced by: overproduction of the chromosomally encoded beta-lactamase or an extended-spectrum beta-lactamase. These activity modification are rarely found in the hospital of Aix-en-Provence (France). The activity modification rate of K. oxytoca resistant by one of these mechanisms between 1986 and 1991 are 3.6% for chromosomally encoded beta-lactamase overproduction and 0.7% for acquisition of an extended-spectrum beta-lactamase. We have determined the isoelectric point (pI) by isoelectrofocusing of the beta-lactamases isolated from 67 K. oxytoca and the activity of 8 beta-lactams has been studied by disk-diffusion. 51 wild strains, 14 overproducing strains (including 2 in vitro mutants) and 2 strains with extended-spectrum beta-lactamase were studied. For every wild strain, we observed only one band except for two strains with two bands (pl 5.4 + 6.3 and pl 5.6 + 7.7). The isoelectric points for the other strains are comprised between pl 5.25 and pl 8.2: 22 pl 7.7; 13 pl 5.25; 4 pl 5.6; 4 pl 6.3; 2 pl 6.6; 2 pl 8.1 and 2 pl 8.2. In the strains with chromosomally encoded beta-lactamase overproduction we observed several bands in each extract and only the major band was considered. The isoelectric point of in vitro mutant strains with beta lactamase overproduction was the same that the wild strains. We observed 3 pl: 5.25 - 1 pl: 5.6 - 3 pl 6.3 and 5 pl 7.7.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233634 TI - [Evaluation of a micromethod for the bactericidal activity of five disinfectants on 108 hospital strains]. AB - The authors studied 5 disinfectants containing aldehyde and quaternary ammonium against 108 hospital strains. The minimal bactericidal activities (MBC) were determined with a microdilution method and the dilution-neutralization method (AFNOR T 72-150). The microdilution method results were validated using the MBCs of four reference strains tested with the dilution-neutralization method (AFNOR T 72-150). 3 disinfectants proved to be valuable. 1 disinfectant was valuable but with a MBC toward all bacteria only a dilution away from the dilution of use. 1 disinfectant was determined as unsatisfactory as 9.5 p. cent of all strains proved resistant toward it. All disinfectants matched AFNOR standards; but those specifications did not match toward hospital strains. Such an evaluation using a simple and reliable method should be of use. PMID- 8233635 TI - [Chlorhexidine digluconate/chlorocresol combination. In vitro study of interactions observed against potential cutaneous strains]. AB - The interaction type of chlorhexidine digluconate/chlorocresol association (foaming solution DC 102/1, Laboratoires Pierre Fabre Medicament) have been studied according to the chess-board method, on microplate, by determining MBC (Minimal Bactericidal Concentrations) (or MFC (Minimal Fungicidal Concentrations)) adjusted to T 72-150 AFNOR norm (dilution-neutralization, contact 5 mn) and computation of FBC (or FFC) Index. Trials were carried out against 18 hospital strains (Staphylococcus epidermidis, Staphylococcus aureus, Propionibacterium acnes, Candida albicans) and against 4 strains of the AF-NOR norm. MBC of chlorhexidine, against Staphylococci were included between 6.25 and 25 micrograms/ml, and between 25 and 200 micrograms/ml against the others strains. MBC of chlorocresol were included between 800 and 3,200 micrograms/ml. Against the 22 tested strains, 18 synergistic interactions (FBC Index < or = 0.75) were observed, with 4 to 8 reducing factor for chlorhexidine, and > or = 2 for chlorocresol. No antagonism was noted (maximal FBC Index = 1). PMID- 8233636 TI - Antiseptic activity of some antidental plaque chemicals on Streptococcus mutans biofilms. AB - S. mutans is one of the bacterial species involved in the initiation of cariogenic dental plaque. In situ, antiseptic activity evaluation of dental plaque chemicals is fairly difficult. A dynamic biofilm model of S. mutans on Tygon support is suitable for the quantification of antiseptic activity and the comparison of commercial chemical formulations, submitted or not to the french standards and Pharmacopea. Some of these chemicals, especially quaternary ammonia solutions, do not reduce living bacteria counts by 10E5 as AFNOR specifies. In the case of dental plaque, antiseptic activity is probably not the only relevant parameter: biofilm dissociation is rather more relevant because the dead or living bacteria adhere to the teeth, favouring the establishment of a new active biofilm. Two proceedings can be followed: support surface can be modified, or bacteria exopolymers synthesis can be inhibited to prevent cell adherence; the disruption of the biofilm glycocalyx can be achieved with enzymes or surfactants. PMID- 8233637 TI - [Experimental study of the antibacterial activity of cloth impregnated with a disinfectant solution]. AB - Disinfecting cloths were commercialized for several years. The solutions of disinfectant used are generally active in vitro in the conditions required by the french standards published by AFNOR, but it is also necessary to know the effectiveness of disinfecting cloths in conditions of use. We describe in this paper a method for this determination. Four bacterial stains recommended in the french standards were used: Staphylococcus aureus ATCC 9144, Enterococcus hirae ATCC 10541, Pseudomonas aeruginosa CIP A 22 and Escherichia coli ATCC 10536. The bacteria were layed on a test watch-glass and the disinfecting cloth was applied on it following the conditions recommended by the manufacturer. 15 minutes after using the disinfecting cloth, the bacteria on the watch-glass and those which were recovered on the disinfecting cloth during application were collected in a neutralising agent suitable for the disinfecting solution used and the surviving bacteria were counted. A similar test was performed using cloth impregnated with sterile distilled water. We studied the activity of unweaved cloths impregnated with a mixture of quaternary ammonium compounds (0.5%) and peroxides (0.1%). The results show that the method allows us to estimate the proper activity of the disinfectant used. In this study, a much higher decontamination of the watch glass was observed with impregnated cloths than with cloths containing distilled water only. The mean reduction of the number of bacteria layed on the watch-glass was 5.95 log10 in 15 minutes in the first case and only 2.4 log10 in the second case. In addition, the disinfectant leads to the destruction of the bacteria recovered on the cloth during application. PMID- 8233638 TI - [Tissue diffusion of amoxicillin-clavulanic acid for antibiotic prophylaxis in colorectal surgery]. AB - Tissue penetration of amoxicillin and clavulanic acid given for antibiotic prophylaxis was studied in 15 patients. On induction of anesthesia patients were given 2 g amoxicillin and 200 mg clavulanic acid. The same dose was given two hours later. At time of surgical incision amoxicillin and clavulanate levels were 15.9 +/- 7.7 micrograms/g and 2.23 +/- 2.69 micrograms/g in abdominal wall fat and 16.6 +/- 10.5 and 1.72 +/- 1.35 micrograms/g in epiploic fat, respectively. Similar levels were measured at time of closure of the abdomen. In colonic wall, amoxicillin et clavulanate levels were 22.7 +/- 13.4 micrograms/g and 2.65 +/- 2.35 micrograms/g. By comparison with blood levels, tissue penetration of both drugs was 20 to 30 p. cent into fatty tissues and 60 to 70 p. cent into colonic wall. PMID- 8233639 TI - [Effect of ketoprofen on the pharmacokinetics of two fluoroquinolones in males]. AB - The influence of a non steroidal antiinflammatory drug (NSAID), ketoprofen, on the pharmacokinetics of two fluoroquinolone derivatives, pefloxacin (P) and ofloxacin (O), was studied in ten healthy adult male volunteers. The subjects were given orally for three days the quinolone alone (P: 400 mg q 12 h and O: 200 mg q 12 h), with at least a one week interval between the two quinolone studies. On day 4, the first kinetic study of pefloxacin and ofloxacin was performed. During the three following days, the quinolone was administered in association with ketoprofen (100 mg daily). Another pharmacokinetic study of P and O was performed on day 8 and the kinetic data obtained were compared to those found on day 4. During the two kinetic studies (D4 and D8), blood samples were taken at times 0, 1, 2, 3, 4, 6, 8, 10, 12 et 24 h and urine was collected during the time periods 0-4 h, 4-8 h, 8-12 h and 12 24 h. Plasma and urine concentrations of the active P and O drug were measured by microbiological assay. Ketoprofen administered for three days with the fluoroquinolone derivative induced no statistical modification in the kinetic parameters of both P and O: peak plasma levels, time to peak level, areas under the curve, apparent volume of distribution, total and renal clearances. PMID- 8233640 TI - Serum and urinary cefpodoxime levels and time killing curves performed in the urine of children presenting urinary tract infections. AB - Since resistance to several oral antimicrobials useful for the treatment of pediatric urinary tract infections (UTI) is overwhelming in Argentina, an in vitro investigation was performed testing 400 isolates obtained from urines of children suffering UTI's, 200 collected in 1990 and 200 in 1991. Their susceptibility against oral antimicrobials marketed in Argentina and appropriate for the treatment of UTI was determined by the agar dilution methods. An increase of the resistance to aminopenicillin combined with beta-lactamase inhibitors and to fluoroquinolones was observed comparing the two periods. Cefpodoxime (CPD), cefixime and fluoroquinolones except norfloxacin were the sole oral antimicrobials showing in vitro activity at the 90 per cent level. Unfortunately fluoroquinolones are not yet approved for pediatric use. Consequently we realized an in vitro and in vivo pharmacokinetic study in order to determine CPD activity against E. coli isolated in UTI cases. Five children (6-10 y) showing E. coli UTI infections received 10 mg/kg/d CPD in a single oral daily dose and were treated up to 10 days, 3 had lower UTI and 2 upper UTI. All patients were clinical and bacteriologically cured. Cultures obtained up to 4 weeks after treatment were negative. CPD serum levels at 2 hours after the first dose of treatment showed a median of 2.7 mg/l (2.3-3.4 range). Bactericidal serum titers at the same time against the patients own strain and an E. coli TEM-1 hyperproducer strain (MIC 4,096 mg/l for ampicillin and 0.5 mg/l for CPD) showed a median value of 1/8 against patients strains and 1/2 against the THP strain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233641 TI - [Mechanisms of adhesion of Staphylococci to biomaterials: effect of fusidic acid]. AB - The production of slime, adherence to plastics and hydrophobicity are factors which regulate the colonisation of biomaterials by Staphylococci. The influence of fusidic acid on these 3 factors was studied by using 3 pairs of pathogenic strains of S. aureus and S. epidermidis. Each pair presented differences in the expression of one or several of these factors. The influence of fusidic acid was initially studied by determining the expression of these factors by these strains cultured in the presence of 0.03 mg/l and 0.5 mg/l of antibiotic. Hydrophobicity was measured by the Bath-test method, slime was detected by Trypan blue staining after fixation with Carnoy's fixative and adherence was determined on polystyrene. The variations observed were generally minor, except for S. epidermidis, high slime produces, which showed a reduced production. Using this collection of strains, we then selected mutants resistant to 2 micrograms/ml of fusidic acid. This resistance induced a reduction in the 3 colonisation factors and it can be proposed that strains resistant to fusidic acid have a lesser capacity to colonise than sensitive strains. The bacteria adhere to and colonise the majority of surfaces proposed to them [3, 7, 10]. This was the case for biomaterials used in medicine, whose number and diversity (catheters, prostheses) are continually increasing. Apart from thrombosis, the major complication is the development of infection. Coagulase positive or negative Staphylococci are very frequently responsible for this type of infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233642 TI - [Quantitative viremia and p24 antigenemia in short-term follow-up of the treatment with zidovudine in twenty HIV infected patients]. AB - The authors emphasize the interest of detecting HIV p24 antigen after immune complexes dissociation as well as changes in cellular viremia titers in the therapeutic follow-up of HIV infected patients. A significant decline of cell virus titers was found in 4-8 weeks among 13 of 20 zidovudine treated patients. A significant decrease of immune complexes dissociated p24 antigenemia was noted in 11 out of 17 patients. PMID- 8233643 TI - Impairment of natural resistance to Toxoplasma gondii infection in rats treated with beta adrenergics, beta blockers, corticosteroids or total body irradiation. AB - The data shows that Fisher rats treated with propranolol, betamethasone or exposed to cobalt60-treatment before challenge with T. gondii exhibit an impaired resistance to this opportunistic parasite infection. In fact, in rats propranolol treatment induces an increase mortality rate up to 66.6% (P < 0.01, with respect to controls). Whereas betamethasone or cobalt60-treatment induces an increased of mortality rate about 33.3% (P < 0.05 respectively, with respect to unirradiated rats). In contrast to beta blocker, corticosteroid-treated or irradiated rats, beta adrenergic-treated rats do not significantly differ from untreated rats in their time of survival. These data lead the authors to hypothesise that in rats the natural resistance to T. gondii can be modulate beta adrenergics or corticosteroids. PMID- 8233644 TI - [Value of filariopheresis in the treatment of Loa loa filariasis]. AB - To prevent the side effects as encephalitis, related with diethylcarbamazine treatment in loiasis, an exchange blood transfusion was proposed. To avoid the disadvantage of this heavy technique, the authors are proposing filariopheresis to trap the microfilariae using cytapheresis technique and filtration. They obtained a medium 75 p. cent microfilariae extraction: 55-70 p. cent by cytapheresis and 5-20 p. cent by filtration. The number of blood cells must be supervised as an important decrease of the platelets may occur temporary. This simple and not expensive technique is efficient, but diethylcarbamazine treatment must be instituted very quickly after filariopheresis session. Furthermore, the millions of microfilariae collected can provide useful antigen for immuno parasitological tests. PMID- 8233645 TI - [Prevalence of Clostridium difficile and toxin A in feces of HIV infected patients]. AB - Patients with AIDS are immunodeficient, receive multiple antibiotic treatments, occasionally anti-cancer chemotherapy and are often hospitalised; thus they are susceptible to develop a Clostridium difficile infection. The aim of this study was to evaluate the role of C. difficile in diarrhoea in this patient population. Therefore, C. difficile and toxin A which plays a major role in pathogenicity were examined in faecal samples of HIV infected patients. Between January 1991 and June 1992, 102 stool samples from 67 patients were studied. Ninety p. cent of these patients were hospitalised (length > 3 days), 80% had a diagnosis of AIDS stage IV, and 66% had diarrhoea. Nineteen point four p. cent of the patients were carriers of C. difficile. Different associations were found: 1) presence of non toxigenic strains and absence of toxin A in stool samples (6 patients), 2) presence of toxigenic strains and absence of toxin A in stool samples (6 patients), 3) presence of toxigenic strains and toxin A in stool samples (2 patients). None of the patients developed a colitis or pseudomembranous colitis. The carrier rate was identical to those found in other hospitalised populations without AIDS. The prevalence of C. difficile diarrhoea or colitis is low. In this study, AIDS patients do not seem to constitute a risk group for C. difficile intestinal pathology. However, carriers of C. difficile were subjected to strict hygiene rules to prevent nosocomial spread. PMID- 8233646 TI - [Survey with three epidemiological markers after 22 cases of diarrhea caused by Clostridium difficile in a geriatric hospital]. AB - In a geriatric hospital of Saint-Etienne (Charite), among 153 patients having presented a nosocomial diarrhea from September 1990 to August 1991 Clostridium difficile (C.d.) has been isolated in 22 cases. Two of the nine units of the hospital had the highest incidence rates: 4.6 and 3.7%. In the faeces of 16 patients, C.d. was toxinogenic. In all cases, except one, antibiotic preceded diarrhea. Amoxicillin + clavulanic acid treatment was the most frequently responsible (65%). For detecting an eventual outbreak, several epidemiologic markers were evaluated: Clindamycin MIC, protein profiles, serotyping. Clindamycin susceptibility differentiated two Cd. types, but has no epidemiologic value. Protein profiles, performed by SDS-Page, individualized 6 different profiles, but 10 strains gave no classifiable profiles. Serotyping, applied by M. Delmee, appeared as the most interesting marker. Inquiry allowed to eliminate an outbreak but revealed two episodes of cross contaminations in the 3 units, 2 of them having the highest incidence rates. Markers proved persistence of the same C.d. strain in some patients who were correctly treated by metronidazole. PMID- 8233647 TI - [Evaluation of the carriage of Staphylococcus aureus in patients and the personnel of a hemodialysis center for the prevention of infections]. AB - With the aim of improving the prevention of Staphylococcus aureus infections in hemodialysis patients, an evaluation of S. aureus nasal carriage was carried out at the hemodialysis center of CHU-Rouen between the 1st of January, 1991 and the 30th of June, 1991. The S. aureus strains were classified according to their antibiotypes, serotypes and lysotypes. The carriage rate appears to be similar to that of the general population but inferior to what has been previously reported in hemodialysis centers. We report our findings on nasal carriage strains. The risk of infection is low. The standards of hygiene, adopted by hospital personnel, seem to be effective although cross colonization may have possibly occurred. Taking into account the different epidemiologic circumstances encountered in each hemodialysis center it is, therefore, necessary to determine the rate of carriage and identify the personnel at risk (persistent carriers, patients with a previous history of S. aureus septicemia, meti-R strains) in order to implement prophylaxis accordingly to epidemic characteristics of each center. PMID- 8233648 TI - [Evaluation of three automated blood culture systems. Bio Argod, Bact T/Alert, bactec NR-860]. AB - In 1991 and 1992, three automated blood culturing systems were successively assessed ("Diagnostics Pasteur" Bio Argos: 1,120 Vials, "Organon Teknika" BacT/Alert: 1,400 vials and "Becton Dickinson" bactec NR-860: 2,146 standard vials 6 and 7 and 2,360 resin vials 26 and 27), and compared with the conventional system (C) (Roche aerobic BHI-S Liquoid, and Diagnostics Pasteur anaerobic prereduced Shaedler). Duplicate blood cultures were collected in various hospital services. They were processed over a 7-day period by the automated instruments and were incubated for 10 days using conventional techniques. The percentage of positivity (clinically significant results) obtained with the automated instruments was similar to that obtained with conventional techniques. Bio Argos: 3.4 p. cent vs 3.8 p. cent with C, BacT/Alert: 3.9 p. cent vs 3.5 p. cent, Bactec standard 6 and 7 vials: 5.6 p. cent vs 6.4 p. cent, and resin 26 and 27: 4.2 p. cent vs 3.1 p. cent. A better sensitivity was found with Bactec resin 26 and 27 vials when compared to C for Staphylococcus aureus (22 p. cent of S. aureus strains were isolated only with resin vials 26 and 27). False positives were as follow: 0.36 p. cent for BacT/Alert, 2.1 p. cent for Bio Argos and 3.5 p. cent for Bactec. The fastest detection rate was observed with the instrument which shakes the vials continuously (BacT/Alert): 73.6 p. cent of positive vials were detected on the day the vials were received at the laboratory. Ninety-four percent of the Bactec resin 26 and 27 positive vials, 90.6 p. cent of the BacT/Alert positive vials, 84 p. cent of the Bactec 6 and 7 positive vials and 73.7 p. cent of the Bio Argos positive vials were detected within twenty-four hours. PMID- 8233649 TI - [Expression of phosphoprotein P53 in pathology]. PMID- 8233650 TI - [Expression of P53 protein in cutaneous melanoma]. AB - P53 protein in cutaneous melanoma. We report the results of an immunohistochemical analysis about the nuclear phosphoprotein P53 expression performed on 48 primary and 10 metastatic cutaneous melanoma in order to assess the prevalence of the expression of mutant P53 protein (m-P53) in this skin tumour. In our study m-P53 was found in about 46% of primary tumours without any significant relationship with the corresponding metastatic lesions. Therefore the P53 count in cutaneous melanoma is not a prognostic marker of tumour spread and aggressiveness. PMID- 8233651 TI - Nucleolar organizer regions in lymphomas: a quantitative study. AB - Using a silver staining technique, nucleolar organizer region-associated proteins (Ag-NORs) have been studied in paraffin sections of 76 non-Hodgkin's lymphomas, five normal lymph nodes, and five <> lymph nodes. The mean number of nucleolar organizer regions per nucleus was 1.19 (SD:0.09) for normal lymphocytes, 3.04 (SD:0.14) for reactive lymph nodes, 2.79 (SD:0.44) for low grade lymphomas, 6.33 (SD:1.58) for intermediate-grade lymphomas, and 10.53 (SD:1.97) for high-grade lymphomas. There were highly significant differences in Ag-NOR counts among the groups (p < 0.001). The Ag-NOR regions were often observed in nuclei in areas where nucleoli themselves were invisible. It is suggested that this method is useful in diagnostic histopathology and in differentiation of the grade of lymphomas. PMID- 8233652 TI - [Concentration of C cells in the thyroid of rats treated with mercaptoimidazole or levo-thyroxine]. AB - Concentration of C cells in the thyroid of rats treated with Mercaptoimidazole or Levo-thyroxine. The aim of the present study was to elucidate the possible functional relationship between follicular and parafollicular cells of the thyroid gland; therefore we have investigated the behaviour of calcitonin producing cells and serum calcitonin concentration in rats both in resting and hyperstimulation conditions of follicular cells. Our results showed that with regard to follicular mass the concentration of the C cells was reduced in the two groups of rats treated compared to control rats. C cell concentration decrease was associated with reduced serum calcitonin concentration. In conclusion C cell activity is independent of TSH and thyroid hormones circulating levels. PMID- 8233653 TI - [Giant cell arteritis, synonym of Horton disease?]. AB - Giant cell arteritis: a synonym of Horton's disease? Biopsies from the temporal artery of 50 patients with Horton disease were evaluated retrospectively by light microscopy and immunohistochemical methods. All patients were adults over 60 years: 28 female, 22 male. Histopathologic study of the biopsies disclosed four main variants most likely related to different stages of temporal arteritis. Only in 22 cases the typical granulomatous giant-cell pattern was observed. However similar pathologic findings occur in several other chronic vasculitis, granulomatous giant cell inflammation being a mark of an advanced stage of the disease rather than a mark of a specific type of arteritis. Therefore <> and <> cannot be considered synonymous, contrary to what usually happens in clinical practice. PMID- 8233654 TI - [Pericardial mesothelioma. Evolution and update in the last decade]. AB - Pericardial mesothelioma: epidemiological updating on the last decade. In the last decade, 109 cases of primitive malignant pericardial mesothelioma, histologically proved and reported in world medical literature have been added to the 206 cases described in a previous article. This illness, although rare, has been found more frequently in Japan (25 subjects) and in USSR (21 subjects). From the table in annex, the following points emerge: a) On average, pericardial mesothelioma tends to develop in fairly young people (compared to pleural or peritoneal mesothelioma). b) There is more proof that asbestos has an harmful effect on pericardial serosa. c) The most frequent clinical diagnoses refer mainly to acute pericarditis, constrictive pericarditis, cardiac tamponade and sometimes to various types of coronary heart disease, which autoptical examination reveals to be due to the presence of an external compression of the coronary arteries by pericardial mesothelioma. d) This illness, perhaps because of greater research and improved diagnostical means seems to be increasing when compared to the previous period (although this does not appear to be the case in our country). PMID- 8233655 TI - [What is the prognostic significance of histomorphology in small cell lung carcinoma?]. AB - What is the prognostic significant of the histomorphology in the small cell carcinomas of the lung? After the WHO classification of the lung cancer (1981), several studies criticized the subdivision of the small cell carcinoma in three sub-types (oat-cell, intermediate cell and combined types). The role of histology in the prognostic predition has been devaluated. In order to verify the prognostic value of the morphology of the small cell types of lung cancer, we performed a multivariate analysis in 62 patients. The survival rate was analytically compared with the following parameters: nuclear maximum diameter, nuclear form, nuclear chromatism, chromatine distribution, presence of nucleolus, evidence of cytoplasm. The results showed that none of these parameters are able to express a prognostic value. According to the recent studies, we think that the small cell carcinoma of the lung is a neoplasia with a multiform histologic pattern. Differences observed in clinical management are not correlate with the morphology, but with other biological parameters still unknown. PMID- 8233656 TI - [Pharmacokinetics of cytochrome P-450]. AB - Pharmacogenetics of cytochromes P-450. Cytochromes P-450 are a large family of enzymes found in all living species whose function is the activation of molecular oxygen which, in turn, will oxidize an organic substrate. They are divided in two groups: one including the constitutive enzymes that intervene in vital processes such as cholesterol synthesis, cholesterol transfer into steroid and sex hormones, prostaglandin synthesis, etc.; the other group including the inducible enzymes, responsible of the metabolism of exogenous substances. Their concentration increases in the presence of specific substrates, like herbicides, cigarette smoke, hydrocarbons, insecticides, etc.. Of the latter group, the genetic polymorphism of two families is described. Family I is involved in the metabolism of polycyclic aromatic hydrocarbons: an allele codifying for a low activity cytoplasmic receptor (autosomic recessive inheritance) and a high affinity one (recessive inheritance) are present. The transformations carried out by the cytochromes P-450 give origin to intermediate reactive products, epoxides, that bonding to nucleoproteins or nucleic acids, can have either toxic or carcinogenic action. Therefore, the subjects with high affinity genes have an increased risk of cancer. This phenomenon, relating to pulmonary cancer, has been demonstrated in cigarette smokers. Family II is the group of greatest pharmacogenetic and clinical interest, since it is responsible of the polymorphism of the response to different drugs, such as halothane, (malignant hyperthermia), ethanol (alcohol intolerance), nitrosamine, (cancer), debrisoquine (hypotension), spartein (excessive uterine contractions). An increased or reduced ability to metabolize specific substances is the consequence: the pharmacological effects can therefore vary very much in the two classes of carriers of different alleles. Possible future applications of these polymorphisms in clinical practice are discussed. PMID- 8233657 TI - [Bone marrow cryptococcosis as the first manifestation of AIDS]. AB - Bone marrow cryptococcosis as initial sign of AIDS. The Authors describe a case of bone marrow cryptococcosis in a patient, female aged 27, in which a toluene induced aplastic anemia was suspected. The mycosis was at first extracerebral and represented the initial manifestation of unexpected AIDS. The Authors discute the symptomatology and the clinical findings leading to the diagnosis and remark the main point of bone marrow evaluation as aid in diagnosing the infective pathology. PMID- 8233658 TI - [Chorioangioma (placental hemangioma): anatomo-pathologic identification and possible clinical relevance]. AB - Chorioangioma (Placental hemangioma): Anatomo-pathological identification and possible clinical importance. Chorioangioma is found in about 1% of all placentas. However, many small tumours are missed in routine examination or easily mistaken for infarcts. Cases of large chorioangiomas may be related with a number of feto-maternal pathological conditions. Here we report a case of chorioangioma with a brief review on the possible several complications related to its occurrence. PMID- 8233659 TI - [Well-differentiated acinic cell carcinoma of the parotid]. AB - Well-differentiated acinic-cell carcinoma of parotid gland. The authors report a case of well-differentiated acinic-cell carcinoma of the parotid gland with an important lymphoid component, whereas in the aspiration a diagnosis of adenolymphoma was made. In our case cytologic examination showed an abundant cellularity, consisting mostly of lymphocytes. Epithelial elements were monomorphic and had a discrete quantity of amphophilic, granular cytoplasm. Such structures sometimes appeared to be in continuity with ductal-type formations, suggesting normal glandular acini of the serous type. Elements oncocytes-like with abundant, basophilic granular cytoplasm were seen also. These cells oncocytes-like, the rich lymphoid component and the presence of little amorphous debris, suggested an adenolymphoma. The differential diagnosis between ACC, adenocarcinoma, and lymphonodal metastasis did not show problems because of the lacking of the cytological atipie. Histologic examination of the surgical specimen did not show problems of differential diagnosis. Morphologic patterns included acinar, tubuloductal, microcystic and follicular structures. Periodic acid Schiff positivity, resistant to diastase, was noted in most serous cells. An acid alcian blue stain was negative within the cell cytoplasm but moderately positive in the microcystic cavities. The final histologic diagnosis was that of a well-differentiated acinic cell carcinoma with a rich lymphoid component. PMID- 8233660 TI - [Histogenetic problems of a "granular cell tumor" in the trachea]. AB - On the possible ontogeny of the cells of a "granular cell tumor" of the human trachea. In a young patient affected with pulmonary tuberculosis during bronchoscopy a "granular cell tumor" of the membranous part of trachea was occasionally evidentied. After pathologic examination, the specimen is analyzed in order to assess the kind of cells that are present in the lesion: the ultrastructural and immunohistochemical study shows Schwann cells which particularly well developed lysosomal complement. PMID- 8233661 TI - Fatal tubular nephropathy following an overdose of theophylline in an infant. AB - A case of fatal acute intoxication in a 4-month and 20-day-old male infant from an overdose of theophylline (Paidomal) drops retard is described. Apart from the usual symptoms caused by theophylline overdose, severe obstructive tubular nephropathy was also observed. PMID- 8233662 TI - [Hypothesis on the Achean disease]. AB - A hypothesis on the Achaean disease. Analyzing the text in the light of the modern medical knowledge, the article traces a profile of the mortal illness described in the first book of Homer's Iliad. The disease was a severe, acute epidemic zoonosis, rapidly bringing death. First, it affected mules and dogs, then it involved men; the infection spread quickly and the number of deaths was high. These elements suggest that the illness occurred among the Achaeans can be identified with glanders, an infectious disease affecting primarily mules and horses that can be transmitted to man by contact with these animals. PMID- 8233663 TI - The opening pitch in the health care reform playoffs. PMID- 8233664 TI - Tuberculosis in infants, children, and adolescents: new dilemmas with an old disease. AB - After nearly three decades of decline, tuberculosis has resurfaced as a major public health problem in the United States. The largest increase in cases is among 25-to-44 year-olds, an age group likely to have children in their households who are at risk of becoming infected. Aggressive interventions must be pursued to interrupt this continuing cycle of transmission. PMID- 8233665 TI - Should there be universal childhood vaccination against hepatitis B? Part 1: A commentary. AB - In 1991, the Immunization Practices Advisory Committee (ACIP) issued recommendations calling for universal immunization of all U.S. infants against hepatitis B. These recommendations have been widely accepted without addressing several considerations including epidemiology of hepatitis B, economic costs, and questions regarding long-term immunity. Both sides in this issue are discussed. PMID- 8233666 TI - Registered nurses' use of universal barrier precautions in the pediatric emergency room. AB - To assess the frequency of Universal Barrier Precautions (UBPs) use by registered nurses in a pediatric emergency room at a large medical center in the southeastern United States, 10 nurses were observed individually for four-hour periods. Observations were noted using the Universal Precautions Assessment Tool (Gauthier, Turner, Langley, Neil, & Rush, in press). Based on 160 opportunities for UBP use, the nurse compliance rate was 71.9%. PMID- 8233667 TI - Incorporating play interventions into nursing care. AB - Play, as a nursing intervention, has often taken a back seat to all the technical skills performed by nurses. Nurses can and should contribute to implementing and designing play interventions. A review of the play literature forms the basis of a case illustration presented to assist the nurse to plan and implement play in the busy hospital setting. PMID- 8233669 TI - Humanitarian award winner. Ida M. Martinson: an outspoken advocate of home care for children with cancer. PMID- 8233668 TI - A developmental approach to childhood safety education. AB - Injuries are the leading cause of childhood mortality in the United States. Both susceptibility to injuries and understanding of safety issues are significantly influenced by a child's developmental level. Knowledge of developmentally based teaching strategies will enable nurses to provide effective safety education to parents and children. PMID- 8233670 TI - Parents and children, war and separation. AB - During Operation Desert Storm, many parents were separated from their children while they served their country. This article describes the behavioral responses of children separated from their mother or father for military duty. PMID- 8233671 TI - High and dry--low and wet: the key to DI and SIADH. AB - The pediatric critical care nurse is often faced with children who have underlying neurologic problems. Anticipating the occurrence of diabetes insipidus (DI) and syndrome of inappropriate secretion of antidiuretic hormone (SIADH) in these children is essential. There is much confusion, however, surrounding recognition and management of these distinctly different although related problems. The key to differentiating these clinical entities may be found in the simple statement, high and dry; low and wet. PMID- 8233672 TI - Lidocaine 2.5%/prilocaine 2.5% EMLA cream. AB - EMLA cream is a topical emulsion composed of prilocaine and lidocaine, produced complete anesthesia of intact skin following application. For the optimum effect, EMLA cream must be applied and covered with an occlusive dressing for 60 minutes prior to the procedure. This limits its use in emergency situations. However, in most situations where procedures are anticipated, EMLA can be applied in advance, including outpatient surgery. Furthermore, if the procedure is a venipuncture, multiple sites should be prepared in case one's initial attempt is unsuccessful. EMLA has been used in children of all ages and provides significant pain relief for venipunctures and lumbar punctures; it is being investigated in children for intramuscular injections and circumcision. PMID- 8233673 TI - Pediatric management problems: upper respiratory infections. PMID- 8233674 TI - Child neglect in home care: weighing legal and ethical issues. AB - Individual cases of child neglect and/or abuse in pediatric home care pose a number of legal and ethical questions for health care providers. Staff members and agencies must carefully consider all legal and ethical issues before arriving at decisions to terminate care due to noncompliance. PMID- 8233675 TI - Pediatric skin care issues for the home care nurse. AB - Home care nurses are faced with increasingly complex nursing care issues regarding pediatric patients in the home setting. Guidelines for prevention, and principles for treatment as it pertains to pressure-related issues and ostomy care are provided in this article. PMID- 8233676 TI - Applying research to practice on a nursing unit. PMID- 8233677 TI - Latex allergy: a threat to you and your patients? PMID- 8233679 TI - Advocating for children of the world. PMID- 8233678 TI - The Gulf War: pediatric emergency room function. AB - PURPOSE: To analyze the effects of war on the civilian population through patterns of medical service consumption in a pediatric emergency room. METHODOLOGY: A survey of variables related to use of emergency room services before, during, and after the Gulf War. RESULTS: There was a dramatic decrease in the number of visits during the war. During the war, there were fewer visits for URI, home accidents and intoxications; there were more visits for complaints such as headaches, restlessness, and tremors. CONCLUSIONS: Visits during the ware were more related to emergency situations with fewer visits of the walk-in clinic type. PMID- 8233680 TI - MRI of the pulmonary artery after arterial switch operation for transposition of the great arteries. AB - Twenty-six children who underwent an arterial switch operation for transposition of the great arteries were examined with MRI and Doppler ultrasound (US). The aim was to determine the capability of MRI for detecting a possible stenosis of the pulmonary arteries. In 10 children angiocardiography (ACG) was done after the MRI examination. Using ACG as the gold standard, MRI was sensitive in the detection of pulmonary artery stenosis. MRI was more precise in the localization of a stenosis than was Doppler US. Right ventricular hypertrophy was a sensitive indicator of pulmonary artery stenosis. These results suggest that MRI could serve as a screening examination for pulmonary artery stenosis after an arterial switch operation. PMID- 8233681 TI - Comparison of sedation regimens for pediatric outpatient CT. AB - The purpose of this study was to compare the concurrent use of chloral hydrate (CH), intramuscular Nembutal (IMN) and intravenous Nembutal (IVN) for sedation. Data was collected on all pediatric outpatients requiring sedation for CT over a 2.5-year period. During this period, 2178 outpatients required sedation of whom, 1324 (60.8%) received IMN, 710 (32.6%) IVN and 110 (5%) CH. The overall success rate was 97% and was similar in all three groups. IVN however, produced better quality sedation than IMN or CH. The IVN group received a significantly lower dose of Nembutal than the IMN group (p = 0.001). Patients receiving IVN had a significantly lower induction time (p = 0.0001) and total examination time (p = 0.001) than IMN or CH. There was an increased occurrence of desaturation in patients sedated with IVN, especially in those with airway obstruction. IVN sedation permits faster, more efficient and better-quality sedation for outpatient CT scanning than IMN or CH in patients requiring intravenous contrast but may not be appropriate for departments that perform relatively few sedations or lack proper resuscitation facilities. PMID- 8233682 TI - Pediatric sedation: short-term effects. AB - A prospective investigation on the short-term effects of various sedation regimens on 549 nonhospitalized magnetic resonance (MR) patients was performed. The drugs evaluated were chloral hydrate, pentobarbital, midazolam, and diazepam (fentanyl was used for enhancement after any of these drugs). The overall safety and efficacy were quite good with all the regimens. Overall, 84% of children slept less than 8 h after the examination, 90% were drowsy and/or unsteady for less than 8 h after they awoke, and 97% resumed their usual activities by 24 h. Significant hyperactivity was seen only with pentobarbital and occurred in 8.4% of children over 8 years of age. The multiple-dose regimen of pentobarbital and fentanyl had a significant short-term effect on the children less than 8 years of age, with 35% sleeping longer than 8 h after the MR. Ten children who had needed the multiple-dose pentobarbital regimen or who had failed prior pentobarbital sedation presented for repeat sedation. Midazolam was effective in 9 of these 10 children. PMID- 8233683 TI - Mucopolysaccharidosis: thickening of dura mater at the craniocervical junction and other CT/MRI findings. AB - Cranial CT and/or MRI imaging of 8 patients with mucopolysaccharidosis (MPS) was retrospectively evaluated. Two patients had MPS IH, 1 had MPS IS, 1 had MPS IVA and 4 had MPS IV. CT and MRI showed thickening of dura mater at the cranio cervical junction, causing narrowing of the subarachnoid space, in all the patients examined. Spinal cord compression was detected in 4 patients. Other findings were: white matter alterations, mild to severe hydrocephalus, skull dysplasia and odontoid dysplasia. White matter alterations were evident as large areas and as multiple dispersed spots of prolonged T1 and T2 value. Reduced gray/white matter contrast was demonstrated on T2-weighted MRI images. It is important to examine the cranio-cervical junction carefully for thickening of dura mater in all patients with mucopolysaccharidosis examined by CT or MRI, because of the generally progressive clinical course of MPS. In patients with symptomatic cord compression, surgical intervention should be considered. PMID- 8233684 TI - Mucopolysaccharidosis I and intracranial tumor in a patient with high-pressure hydrocephalus. AB - In a 19-month-old patient with mucopolysaccharidosis I (Pfaundler-Hurler, MPS I/H) high-pressure hydrocephalus required the implantation of a ventriculo peritoneal shunt. Despite a reduction in both ventricular volume and intracranial pressure, clinical symptoms suggesting compression of the brain stem persisted. Brain MRI revealed a tumor within the posterior cranial fossa. Cytologic examination of the cerebrospinal fluid was suggestive of a poorly differentiated ependymoma. High-pressure hydrocephalus is a common complication in MPS I/H. As changes in mucopolysaccharide metabolism may be associated with an increased risk of developing neoplasms, the possibility of an intracranial tumor should be considered in patients with MPS I/H and high-pressure hydrocephalus. PMID- 8233685 TI - Radiographic characteristics of isolated invaginated Meckel's diverticulum. AB - A 3-year-old boy with a radiographic finding of an isolated invaginated Meckel's diverticulum is presented. The abnormality simulates a polypoid filling defect in the distal small bowel on barium examination. This particular manifestation of Meckel's diverticulum is very rare and has been reported only once previously. This article re-emphasizes the need to think about this possibility when a polypoid filling defect is seen in the distal small bowel. At the time laparotomy was performed, the abnormality had progressed into a triple intussusception, a rare surgical finding. PMID- 8233686 TI - Gallbladder pathology in pediatric beta-thalassemic patients. A prospective ultrasonographic study. AB - Eighty-three beta-thalassemic patients, aged 1.5-19 years, were prospectively studied for evidence of gallbladder pathology by real-time, high-resolution ultrasound. Cholelithiasis was present in 7 patients (8.4%). Sludge, a clinical entity that, when it persists, can be a predisposing factor for cholelithiasis and cholecystitis, was detected in 29 patients (34.9%). Prevalence of sludge to this extent has not been reported in the literature and we believe this is due to better technology offered by modern ultrasound equipment. Pseudo-sludge, a condition that is created by a beam averaging effect (partial-volume phenomenon), at the diverging portion of the ultrasound beam, has always been confusing and a source of difficulties in the ultrasound examination of the gallbladder. For the first time, a method is described, to easily differentiate between true sludge and pseudo-sludge. PMID- 8233687 TI - Retrograde jejunal-duodenal intussusception associated with a jejunal duplication cyst in a newborn. AB - Retrograde intussusception is a rare occurrence. We report a retrograde jejunal duodenal intussusception associated with a jejunal duplication cyst, which presented as intermittent biliary and upper gastrointestinal (UGI) obstruction and a mobile abdominal mass in a newborn infant. PMID- 8233688 TI - Sonography of the circumrenal and horseshoe adrenal gland in the newborn. AB - The adrenal gland can be visualized using ultrasonography in at least 90% of fetuses of more than 26 weeks gestation. The fetal and neonatal adrenal gland is described as a structure having a characteristic V or Y shape. Despite the frequency of visualization of the neonatal adrenal gland, few reports of adrenal anomalies are present in the radiologic literature. The purpose of this paper is to describe and depict the sonographic appearance of the "horseshoe" and the "circumrenal" adrenal gland and to discuss adrenal anomalies in general. PMID- 8233689 TI - Duplex Doppler examination of renal allografts in children: correlation between renal blood flow and clinical findings. AB - We reviewed the diastolic blood flow (DBF), pulsatility index (PI), and resistive index (RI) in 211 duplex Doppler ultrasound examinations of 63 pediatric renal transplant recipients. We correlated our findings with the clinical diagnosis, independently determined at the time of ultrasound examination. DBF, evaluated in 202 examinations, was found to be decreased, absent, or reversed in acute rejection (kappa = 0.4, p < 0.01) and acute tubular necrosis (ATN) (kappa = 0.3, p < 0.01). The PI and RI were evaluated in 87 and 78 ultrasound examinations respectively. The range of normal PI and RI values in recipients between six and 18 years of age were similar to those reported in adults. However, normal PI and RI values were somewhat higher in recipients less than six years of age who had received adult donor allografts. Overall, the PI was greater than 1.5 in acute rejection (kappa = 0.3, p < 0.05) and ATN (kappa = 0.3, p < 0.01), but not in chronic rejection (kappa < 0, p = NS). The RI was greater than 0.7 in chronic rejection (kappa = 0.3, p < 0.05) and ATN (kappa = 0.3, p < 0.05), but not in acute rejection (kappa = 0.03, p = NS). We conclude that the PI and RI vary in pediatric recipients of renal allografts and are not associated with a specific clinical diagnosis. PMID- 8233690 TI - Vaginal occlusion induced by cancer therapy. AB - Two adolescent girls with acquired vaginal occlusion and resultant hematometracolpos are presented. Both are long-term survivors of childhood malignancies, who prior to multiagent chemotherapy and pelvic irradiation had normal uteri by imaging. Both were amenorrheic post therapy. Routine follow-up computed tomography (CT) scans to monitor their malignancies demonstrated unexpected cystic pelvic masses, subsequently diagnosed as hematometracolpos. PMID- 8233691 TI - Acute urinary retention in an infant: an unusual presentation of a paraureteral diverticulum. AB - We present a case of a male infant with no previous urological symptoms who presented in acute urinary retention caused by compression of the posterior urethra by a paraureteral diverticulum. We discuss the etiology, evaluation and management of bladder outlet obstruction secondary to a paraureteral diverticulum in children. PMID- 8233692 TI - Surfactant protein B deficiency: radiographic manifestations. AB - Surfactant is a complex structure primarily composed of phospholipids, but containing essential proteins as well. Congenital deficiency of Surfactant Protein-B (SPB) has recently been documented for the first time in two siblings. The pathologic findings in these infants was that of congenital pulmonary alveolar proteinosis and the radiographic manifestations were strikingly similar to hyaline membrane disease. PMID- 8233693 TI - High-resolution CT in the assessment of bronchiectasis in children. AB - To assess the possibilities and limitations of high-resolution CT (HRCT) in the evaluation of bronchiectasis in children, we conducted a prospective study of 20 children with clinical and/or chest film findings suggestive of this diagnosis. The 2-mm collimation, 4.3-s HRCT scans with 10 mm interslice spacing were obtained in areas of suspected bronchiectasis; in nonsuspect areas 25-30 mm interslice spacing was used. No preparation for examination was required. Bronchiectasis was revealed in ten patients (50%), being bilateral in four cases and unilateral in six cases. All types of bronchiectatic patterns were found. Cooperation during the examination was the only difference when compared with an investigation of adults. It was not a serious problem in children aged 7 years and older; scans in 6-year-old children were diagnostic but not ideal. Nondiagnostic scans were obtained in a 3-year-old girl. At the time of the scans only one patient had undergone surgery. Preoperative bronchography confirmed the CT findings. The authors conclude that HRCT can limit the need for bronchography in children with a CT finding of focal bronchiectasis in whom surgery is contemplated. When using longer scanning times it is not possible to obtain good results without sedation of children younger than 6 years. PMID- 8233694 TI - High resolution CT of the lungs in acute disseminated tuberculosis and a pediatric radiology perspective of the term "miliary". AB - High resolution CT (HRCT) of the lungs in six children with acute disseminated tuberculosis was evaluated. There was a wide variation in the HRCT appearances. This covered differences in size, distribution and concentration of nodular opacities. Coalescence of nodules and the presence of interstitial thickening was variable. The recognition of acute disseminated tuberculosis is important for diagnosis and has prognostic implications. The use of the term "acute disseminated tuberculosis" rather than "military tuberculosis" is advocated. PMID- 8233695 TI - Pericardial effusion and cardiac hemangioma in the neonate. AB - In the absence of hydrops or sepsis, a pericardial effusion is a rare occurrence in the neonate. We report a case of a neonate with a pericardial effusion in which there was an associated intracardiac hemangioma. Our literature review found 32 cases of pericardial effusion without hydrops in infants under 3 months of age; twelve of these cases were associated with intracardiac and pericardial tumors; 20 others were discovered to be randomly associated with other problems. PMID- 8233696 TI - Bubbly intrapleural mucous simulating congenital lobar emphysema. AB - A neonate presented for an esophageal atresia repair and developed respiratory distress in the immediate post-operative period. The initial conventional chest radiographs suggested that there was a right sided pneumothorax; however, this did not resolve, even after the placement of two chest tubes. A CT examination suggested that the findings were due to hyperinflation of the right upper lobe. Thoracoscopy was performed and revealed a complex pneumothorax composed of bubbly mucous from a post-operative pleuro-esophageal fistula. PMID- 8233697 TI - Significance of periportal low-attenuation zones following blunt trauma in children. AB - The CT scans of 400 consecutive children evaluated with CT following blunt abdominal trauma were evaluated to determine the frequency of periportal low attenuation zones, assess patterns of associated intraabdominal injury, and examine clinical outcome. Periportal low-attenuation zones were noted in 60 children (15%). The presence of these zones was associated with a significantly higher incidence of intraabdominal injury (60% versus 11%, p = 0.0001). Injuries most frequently associated with zones of periportal low-attenuation included hepatic (n = 23, 38%), and adrenal (n = 14, 23%). Children who had periportal low attenuation zones tended to be more physiologically unstable as evidenced by a lower Trauma Score (diffuse, 11.9; focal, 13.4) than children without the zones (15.1, p = 0.0001). The presence of these zones was also associated with a significantly higher mortality rate (13% versus 1%, p = 0.0001). Ten children who had periportal low-attenuation zones and no hepatic injury on CT had a normal appearing liver on gross inspection at surgery or autopsy. In conclusion, periportal low-attenuation zones are common in children who have hepatic injury. These zones may be seen in conjunction with non-hepatic visceral injury or in the absence of intraabdominal injury. The presence of zones of periportal low attenuation is associated with a higher index of physiologic instability, and higher mortality. PMID- 8233698 TI - Urokinase treatment of forearm ischemia complicating supracondylar fracture of the humerus in three children. PMID- 8233699 TI - Cranial ultrasound findings in aspartoacylase deficiency (Canavan disease). AB - Canavan disease (CD) is a rare leukodystrophy which is lethal in infancy or early childhood. The underlying biochemical abnormality in CD is a hereditary deficiency of N-aspartoacylase transmitted in an autosomal recessive fashion. We report on the ultrasound (US), CT, and MRI findings of three unrelated boys with biochemically confirmed CD. At 6 and 9 months of age, two CD patients with rapid neurological deterioration showed markedly enhanced acoustic attenuation of the white matter with the exception of the corpus callosum, giving the appearance of a reversed pattern of echogenicity of cortical gray and subcortical white matter. While gyri and sulci had an almost normal US appearance, the periventricular gray matter featured prominently with increased echogenicity. In contrast another CD patient with a more protracted course had ventricular enlargement when examined by US at 5 and 9 months but no alteration in white matter echogenicity. MRI showed impaired myelinization in all three patients with Canavan disease. PMID- 8233700 TI - Cranial fasciitis of childhood. AB - We present a child with a rapidly growing mass and lytic skull lesion that on pathologic evaluation was diagnosed as cranial fasciitis. This disease entity is not widely known by radiologists, and should be included in the differential diagnosis of lytic skull lesions. PMID- 8233701 TI - Adrenoleukodystrophy. A case report demonstrating unilateral abnormalities. AB - Adrenoleukodystrophy is a hereditary disorder characterized by progressive demyelination of cerebral white matter and adrenal insufficiency. Typical CT and MRI findings in the brain have been documented recently and consist of bilateral white matter abnormalities. We report the case of an 8-year-old boy whose CT and MRI scans showed unusually florid unilateral abnormalities. PMID- 8233702 TI - CT of diffuse leptomeningeal metastasis from primary extracerebral neuroblastoma. AB - Leptomeningeal metastasis due to primary extracerebral neuroblastoma is rare. Diffuse leptomeningeal contrast enhancement resulting from this condition has been demonstrated only in a recent case report utilizing cranial CT imaging. The case presented here is the second example of diffuse leptomeningeal metastasis from primary extracerebral neuroblastoma. Metastasis was visualized by pre- and postcontrast cranial CT scans 6 months after the diagnosis of the disease when the patient had developed multiple cranial nerve palsies. PMID- 8233703 TI - Inadvertent transcervical catheterization of the uterus in an infant with intraperitoneal injection of a contrast medium. AB - A case is presented in which attempted catheterization of the urethral meatus resulted in the inadvertent catheterization of the cervical os. Visualization of the urethral meatus is a requisite for successful catheterization of infant females and young girls. PMID- 8233704 TI - Aneurysmal bone cyst in early childhood. Ultrasound findings. AB - A 1-year-old child with an aneurysmal bone cyst in the scapula is presented. The sonographic examination showed a cystic formation with intraosseal fluid levels that shifted after repositioning of the child. Similar findings have been reported on CT scans and it is suggested that shifting fluid levels are an indicative feature of aneurysmal bone cyst. PMID- 8233705 TI - Clear cell sarcoma of the kidney. A case report. AB - Clear cell sarcoma of the kidney, a tumor which can resemble Wilms' tumor on imaging studies, possesses a much worse prognosis because of its skeletal metastatic pattern. A case of such a tumor is described here. PMID- 8233706 TI - Detection of a deep lipoblastoma by MRI and ultrasound. AB - Lipoblastoma is a rare benign tumor of the soft tissue occurring predominantly during the first two years of life. We report here the application of MRI and ultrasound to the detection of a soft tissue lipoblastoma. By MRI and ultrasound, we precisely evaluated the extent of the tumor, the presence of atypical areas of cystic and mucoid degeneration within the tumor, and its lack of vascularity. Our patient's lipoblastoma did not demonstrate the signal characteristics of a fatty tumor on MRI. PMID- 8233707 TI - Postnatal growth of fetus-in-fetu. AB - A 6-month-old-boy who presented with an abdominal mass disclosed a well-formed fetus in the retroperitoneum. This mass was retrospectively noted at the age of 1 month. Comparison of two radiographic examinations done 5 month apart clearly demonstrated the enlargement of an included fetus in the bearer's abdomen. Our report of this rare condition focuses on the growth of the fetus-in-fetu detected by roentgenograms. PMID- 8233708 TI - Smooth muscle tumors in immunocompromised (HIV negative) children. AB - Two cases of smooth muscle tumor developing in immunocompromised children are described. Both children presented after transplant surgery, one with a hepatic leiomyoma and the other with a hepatic leiomyosarcoma. PMID- 8233709 TI - Muscle carnitine repletion by long-term carnitine supplementation in nephropathic cystinosis. AB - The renal tubular Fanconi syndrome of children with nephropathic cystinosis causes plasma and muscle carnitine depletion. L-Carnitine replacement therapy for up to 18 mo has previously been shown to normalize plasma but not muscle carnitine levels. We treated six cystinosis patients, aged 1 to 4 y, with a mean dosage of 92 mg L-carnitine/kg/d given every 6 h for an average of 62 mo. Despite fractional excretions of free carnitine ranging from 55 to 108%, plasma-free and total carnitine concentrations were maintained at or above normal levels. At the end of the carnitine replacement period, the six children had muscle-free carnitine values ranging from 16.0 to 28.0 nmol/mg noncollagen protein compared with values of 3.0 to 11.4 for cystinosis children not supplemented with carnitine [normal, 22.7 +/- 5.0 (SD) nmol/mg protein]. Total muscle carnitine values were also normalized by L-carnitine replacement. The monthly increase in total body creatinine production, a measure of muscle mass, was higher (p = 0.036) in children with normal plasma free carnitine concentrations (3.4 +/- 0.9 mg/d) than in children with low plasma free carnitine (2.3 +/- 0.7 mg/d). No serious side effects, such as severe diarrhea, were observed. We conclude that oral L-carnitine replacement can normalize muscle carnitine content in children with cystinosis. PMID- 8233710 TI - Effects of endothelin on renal function in newborn rabbits. AB - The renal effects of endothelin-1 were investigated in 16 anesthetized and mechanically ventilated newborn rabbits. Renal blood flow and glomerular filtration rate were determined by the clearance of para-aminohippuric acid and inulin, respectively. Each animal acted as its own control. In eight newborn rabbits, a bolus injection of 5 nmol.kg-1 of endothelin-1 caused an initial fall in mean arterial blood pressure followed by a gradual, significant increase in mean arterial blood pressure that lasted for 45 min. The dramatic increase in renal vascular resistance (+28 +/- 4%) induced by endothelin led to a fall in glomerular filtration rate (-12 +/- 4%) and renal blood flow (-16 +/- 3%). In spite of the reduction of glomerular filtration rate and renal blood flow, urine flow and sodium excretion rates increased significantly (+20 +/- 5% and +49 +/- 9%, respectively). In eight additional newborn rabbits, a bolus injection of 1 nmol.kg-1 of endothelin--a dose that usually induces marked renal and systemic vasoconstriction in adult models--did not affect systemic or renal hemodynamics. In conclusion, endothelin induces renal and systemic vasoconstriction and affects water and sodium homeostasis during the neonatal period. These effects occur under higher doses than those used in adult animals. This age difference in systemic and renal responsiveness is probably mediated by receptor immaturity and/or interference of high levels of counteracting hormones present during the neonatal period. PMID- 8233711 TI - Developmental regulation of the alpha 1B-adrenoceptor in the sheep kidney. AB - The expression of renal alpha 1B-adrenoceptor (alpha 1B-AR) mRNA was studied and contrasted with the expression of renal renin mRNA in fetal and newborn sheep. Fetal sheep between 90 and 91, 116 and 118, and 139 and 141 d gestation (term is 145 d gestation) as well as newborn lambs between 1 and 2 d old and 8 and 10 d old were studied (n = 3 for each age range). The role of the renal nerves in regulating changes in alpha 1B-AR gene expression was also investigated by measuring renal cortical alpha 1B-AR mRNA levels and receptor kd and maximum number of binding sites in 24-h-old lambs that were either denervated (n = 6) or sham-operated (n = 5) 3 d before birth. During development, renal alpha 1B-AR mRNA levels show a marked increase in term fetuses; this increase persists into the first 2 d of life and is distinct from the developmental pattern seen for renal renin mRNA levels. Denervation of term fetuses does not alter the expression of renal alpha 1B-AR mRNA in newborn lambs when compared with sham operated controls but decreases significantly the expression of the renin gene (p < 0.05). These results suggest that the alpha 1B-AR gene is developmentally regulated in the kidney in a pattern distinct from that seen for renin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233712 TI - Decreased plasma ubiquinone-10 concentration in patients with mevalonate kinase deficiency. AB - Patients with mevalonate kinase deficiency suffer from psychomotor retardation, ataxia with progredient cerebellar atrophy, and myopathy. The pathophysiology of the disease remains unclear. The mevalonate kinase product, cholesterol, is within the normal range in patient plasma and fibroblasts. In search of the pathophysiology of this disorder, another mevalonate kinase product, ubiquinone 10, was studied. The concentrations of ubiquinone-10 in patient plasma (n = 6) and ubiquinol-10 in patient LDL (n = 2) and the synthesis of ubiquinone-10 in patient fibroblasts (n = 4) were determined. After oxidative modification of LDL by copper in vitro, the concentrations of alpha-tocopherol and polyunsaturated fatty acids in LDL and the relative electrophoretic mobility of LDL were measured to determine the antioxidant capacity of LDL samples of two affected siblings. The ubiquinone-10 concentrations in plasma samples (median = 508 micrograms/L, range = 488-642 micrograms/L) versus controls (median = 613 micrograms/L, range = 564-809 micrograms/L; p < 0.005) were decreased. In LDL samples of two affected siblings, the concentration of ubiquinol-10 and the resistance to oxidation in vitro were found decreased during intercurrent patient crisis condition. In patient fibroblasts (median = 533 dpm/mg protein, range = 399-1,047 dpm/mg protein) versus controls (median = 40,731 dpm/mg protein, range = 12,774-54,739 dpm/mg protein), the synthesis of ubiquinone was found to be decreased. We conclude that mevalonate kinase deficiency leads to a decreased synthesis of ubiquinone-10 and that ubiquinone-10 deficiency is responsible for the clinical progression of this disease characterized by increased lipid peroxidation, cerebellar atrophy, cataract development, and myopathy with increased creatine kinase activity. PMID- 8233713 TI - An estimate of fetal autonomic state by spectral analysis of fetal heart rate fluctuations. AB - The assessment of the functional state of the autonomic nervous system (ANS) in real time, by means of spectral analysis of fetal heart rate variability, may serve to improve the diagnosis of pathologic conditions of importance to the perinatologist. The combination of two approaches, namely an efficient method for detecting fetal ECG from the abdominal maternal signal, followed by spectral analysis of heart rate variability, is tested as a new noninvasive tool to assess fetal viability in real time. This study demonstrates a pattern of ANS development via the spectral contents of heart rate variability. It is shown that during "quiet state," the "young" fetuses (gestational age = 23.5 +/- 1 wk) present twice as much power of heart rate fluctuations at all frequencies from 0.2 to 1.0 Hz as "mature" fetuses (gestational age = 39.75 +/- 1.5 wk). This finding is coherent with the evolution of a stable and mature ANS activity. At frequencies below 0.1 Hz, a 1/f alpha power law relationship (alpha = 0.85, r2 > 0.9) between spectral density and frequency is displayed in the two age groups. A respiratory peak has been observed in some of the short (64-s) traces we analyzed. However, no respiratory peak was ever observed in a long (256-s) trace, due to the episodic nature of the fetal breathing and immaturity of the ANS. PMID- 8233714 TI - Time-dependent changes in the chronotropic response to vagal stimulation in the newborn canine. AB - We characterized changes in the vagal chronotropic response during 3-min trains of vagal stimulation at 3, 5, and 8 Hz in anesthetized, chemically sympathectomized (6-hydroxydopamine) newborn canines (< 15 d of age). In response to vagal stimulation, the sinus cycle length gradually increased (within 30 s) to a maximum value that was dependent upon the stimulation frequency (p < 0.001). The chronotropic response then attenuated over the remainder of the vagal train. However, unlike in adult dogs, the degree of attenuation of the vagal chronotropic response (fade) was also highly dependent upon the frequency of vagal stimulation in the range 3-8 Hz (p < 0.002). We then compared the maximum change in sinus cycle length and fade in a group of neonates while stimulating the vagus at 3 Hz before and after the administration of physostigmine (0.2 mg/kg i.v.). Physostigmine resulted in a significant increase not only in the maximum percent change in sinus cycle length but in the magnitude of attenuation of the vagal response as well. Therefore, unlike the adult, in the newborn the magnitude of both the vagal response and fade are dependent upon concentrations of acetylcholine released in response to modest levels of vagal stimulation (< or = 8 Hz). This may be the result of differences between the newborn and the adult in the prestreceptor mechanisms of fade or in the susceptibility of the muscarinic receptor to desensitization by the neurotransmitter acetylcholine. PMID- 8233715 TI - The relationship between bradycardia, apnea, and hypoxemia in preterm infants. AB - The pathogenesis of bradycardias in preterm infants is poorly understood. Because their pathogenesis may involve both apnea and hypoxemia, we set out to analyze the proportion of bradycardias that were associated with an apneic pause and/or a fall in arterial oxygen saturation (SaO2), and the temporal sequence of the three phenomena, in overnight tape recordings of SaO2 (Nellcor N100 in beat-to-beat mode), breathing movements, nasal airflow, and ECG in 80 preterm infants at the time of discharge from hospital. A bradycardia was defined as a fall in heart rate of > or = 33% from baseline for > or = 4 s, an apneic pause as a cessation of breathing movements and/or airflow for > or = 4 s, and a desaturation as a fall in SaO2 to < or = 80%. A total of 193 bradycardias were found in 46 (58%) of the recordings (median, three per recording; range 1-18). There was a close relationship between bradycardias, apneic pauses, and desaturations: 83% of bradycardias were associated with apneic pauses and 86% with desaturations. Where all three phenomena occurred in combination, the time from the onset of apnea to the onset of the fall in SaO2 was shorter (median interval, 0.8 s; range -4.9-+ 11.5 s) than that from the onset of apnea to the onset of bradycardia (median, 4.8 s; range -4.0-+ 14.0 s). Hence, most bradycardias (86%) commenced after the onset of the fall in SaO2. We conclude that bradycardia, apnea, and hypoxemia are closely linked phenomena in preterm infants. PMID- 8233716 TI - Abolition of fetal breathing movements by spinal cord transection leads to reductions in fetal lung liquid volume, lung growth, and IGF-II gene expression. AB - Fetal breathing movements (FBM) are considered necessary for normal growth and structural maturation of the fetal lung, but the underlying mechanisms are unclear. The small fluctuations in lung dimensions caused by FBM have been proposed as a stimulus to lung growth, but it is equally possible that FBM act by maintaining the basal level of lung luminal volume, which is an established determinant of fetal lung growth. Our aim, therefore, was to determine the effects of abolishing FBM, while retaining the integrity of the diaphragm, on the volume and rate of production of fetal lung liquid, gene expression for IGF-II, and fetal lung growth. FBM were abolished in seven fetal sheep by high spinal cord transection at 114 +/- 1.2 d of gestation; seven intact fetuses served as controls. At 119 to 124, 125 to 130, and 131 to 136 d, we measured the volume and secretion rate of lung liquid by dye dilution. At these three age ranges, the lungs of cord-transfected fetuses contained 27 to 53% less lung liquid than controls (p = 0.004), and their rates of secretion were 65 to 138% greater (p = 0.001). At postmortem (135 +/- 0.1 d), the lungs of the cord transected fetuses contained less DNA per kg body weight and tended to be lighter and to contain less protein than controls. IGF-II gene expression in the lungs of cord transected fetuses was significantly less than that in controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233717 TI - Distribution of exogenous surfactant in rabbits with severe respiratory failure: the effect of volume. AB - The transient effect of surfactant therapy that is observed in some patients might, at least in part, be explained by a nonhomogeneous distribution. Therefore, we investigated the distribution of a surfactant preparation (Alvofact, 45 g/L) that is used clinically. Rabbits with severe respiratory failure were treated with this surfactant at a dose of 100 mg/kg body weight, and the distribution of surfactant was determined by the use of 141Ce-labeled microspheres that were mixed with the surfactant. Fifteen min after surfactant administration, the rabbits were killed, and the lungs were removed and divided into 200 pieces. The radioactivity per mg lung tissue was determined in each piece. We found that the endotracheal instillation of this surfactant preparation results in a nonhomogeneous distribution. However, a significantly improved distribution was obtained when this dose of surfactant (100 mg/kg body weight) was diluted with normal saline to a concentration of 6.25 g/L. The consequence of the administration of this dose was an intratracheal fluid administration of 16.0 mL/kg body weight. The distribution was also nonhomogeneous after the administration of a small-volume (2.4 mL/kg body weight), low-concentration surfactant preparation (6.25 g/L). We conclude that a surfactant preparation with clinical application is distributed nonhomogeneously in the lungs after endotracheal administration. The distribution can be significantly improved by increasing the fluid volume in which the surfactant is suspended. PMID- 8233718 TI - Interleukin-8 concentrations are elevated in bronchoalveolar lavage, sputum, and sera of children with cystic fibrosis. AB - Concurrent pulmonary inflammation and neutrophil infiltration are characteristic of children with cystic fibrosis (CF). The production of the major neutrophil chemotactic cytokine IL-8 by alveolar macrophages or other cells could be of great importance in the pathology of acute lung disease, but its role in the persistent lung inflammation characteristic of CF has not been evaluated. In this study, we have measured, by ELISA, the concentration of IL-8 in sputum, bronchoalveolar lavage, and sera specimens obtained from children with CF. For comparison, IL-8 in bronchoalveolar lavage obtained from asthmatic patients and from non-CF children with or without lung infection and in sera from age-matched controls was measured. High levels of IL-8 were measured in sputum (mean = 2952 pM) and in bronchoalveolar lavage (mean = 6624 pM) from CF patients. In both cases, there was a significant correlation between clinical status (Schwachman score) and IL-8 levels. This was not true for IL-8 levels measured in sera, which nevertheless were significantly higher in CF patients (p = 0.0001) than in normal controls in the over-10-y age group. PMID- 8233719 TI - Circulating tumor necrosis factor-alpha levels and lipid abnormalities in patients with cystic fibrosis. AB - Hyperlipidemia is prominent among the disturbances in intermediary metabolism that occur subsequent to infections by microorganisms. The response to such infections is known to involve several cell types and is mediated by cytokines. We hypothesized that metabolic lipid disturbances seen during infection in cystic fibrosis (CF) patients may partly be the result of excessive tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine known to cause a large spectrum of pathophysiologic alterations, including impaired lipid metabolism. Therefore, we determined the circulating concentration of TNF-alpha and analyzed its relationship to lipid and lipoprotein levels, as well as lipoprotein lipase activity, in 31 CF patients. Plasma TNF-alpha values were significantly (p < 0.01) elevated in patients with CF compared with controls. The CF subjects were found to have decreased plasma cholesterol (25%), LDL cholesterol (35%), and HDL cholesterol (19%) concentrations, whereas plasma triglycerides were significantly increased (p < 0.001). The apo A-I level was reduced (p < 0.005), whereas apo B levels were normal. Low levels of the major essential fatty acids were found in the plasma of the CF patients, and the triene/tetraene ratio confirmed their essential fatty acid deficiency. Postheparin lipolytic activity was lower in CF patients than in controls, and the decreased activity was accounted for primarily by a decline in hepatic lipase. A significant positive correlation (p < 0.001, r = 0.70) was found between TNF-alpha and plasma triglyceride levels. However, no association was noted between TNF-alpha and essential fatty acid, cholesterol, or lipoprotein cholesterol levels, or with lipoprotein lipase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233720 TI - Immunoprophylaxis and immunotherapy of respiratory syncytial virus-infected mice with respiratory syncytial virus-specific immune serum. AB - The effects of passive respiratory syncytial virus (RSV) immune serum given as preinoculation prophylaxis, postinoculation prophylaxis, and as therapy on d 5 after inoculation were evaluated in an illness model of RSV infection in BALB/c mice. Pre- and postinoculation prophylaxis reduced RSV replication in lung after primary RSV infection and prevented illness. Day 5 treatment did not affect peak titer of RSV in lung but resulted in more rapid recovery from illness than seen in untreated mice. Prophylaxis prevented antibody responses and lymphocytic infiltrates in lung after primary RSV infection. Treatment caused diminished antibody and pathologic responses. Prophylaxis increased susceptibility to reinfection, although illness after rechallenge was mild. Mice treated therapeutically were less susceptible to reinfection than mice treated prophylactically, but they also experienced mild illness after rechallenge. Passive antibody prophylaxis and treatment of RSV infection are promising approaches to attenuating lower respiratory tract illness from primary RSV infection. The ability to measure illness endpoints and pathology make the BALB/c mouse model of RSV infection a useful system for the preclinical evaluation of immunoprophylactic and immunotherapeutic modalities. PMID- 8233721 TI - Simultaneous influence of blood pressure, PCO2, and PO2 on cerebral blood flow velocity in preterm infants of less than 33 weeks' gestation. AB - In extremely preterm infants, the protective capacity for cerebral blood flow (CBF) autoregulation may be impaired or absent, which increases the risk for developing cerebral lesions. The purpose of this study was to quantify the simultaneous influence of several vital parameters, such as mean arterial blood pressure (MABP), PCO2, and PO2, on cerebral blood flow velocity (CBFv), which is used as a measure for CBF. In 16 mechanically ventilated infants of < 33 wk gestation, the CBFv in the internal carotid artery was measured every minute for 1 h by a computer-controlled pulsed Doppler device. MABP and transcutaneous PCO2 and PO2 were recorded as well. A multiple linear regression analysis was performed in each patient to determine the individual MABP, PCO2, and PO2 reactivities as a measure for CBF autoregulation. The medians (and ranges) of the whole group were an MABP reactivity of 7.5% (-12.5 to 20.1%) rise in CBFv/1 kPa rise in MABP, a PCO2 reactivity of 32.7% (-8.1 to 79.5%) rise in CBFv/1 kPa rise in PCO2, and a PO2 reactivity of -3.1% (-14.2 to 7.9%) fall in CBFv/1 kPa rise in PO2. In preterm infants, the individual's capacity for MABP-, PCO2-, and PO2 dependent CBF autoregulation can be estimated by means of the present method, even if the vital parameters change simultaneously. PMID- 8233722 TI - The influence of indomethacin on the autoregulatory ability of the cerebral vascular bed in the newborn lamb. AB - Prevention of hyperperfusion of the brain in the perinatal period has been thought to be an important mechanism by which indomethacin reduces the risk for severe periventricular-intraventricular hemorrhage. The present study investigated whether an indomethacin-induced enhancement of the upper limit of cerebral vascular autoregulatory ability in the neonate contributed to this reduction in cerebral blood flow. In seven anesthetized newborn lambs, we measured temporal blood flow velocity (TMFV) in the carotid artery over a wide range of mean aortic blood pressures (MABP) before and 30 min after an i.v. dose of 1 mg/kg indomethacin. TMFV in the carotid artery was used as an estimate for changes in cerebral blood flow. Stepwise changes in MABP of approximately 10 mm Hg were achieved by progressive balloon occlusion of the thoracic aorta or by progressive bleeding. Multiple linear regression analysis of TMFV versus MABP, indomethacin, and the possible interactive effects confirmed that, at MABP values up to 86 mm Hg, indomethacin lowered TMFV of the carotid artery. Above 86 mm Hg, indomethacin reduced the slope of the TMFV-MABP relationship, indicating an improvement of the autoregulatory ability of the cerebral vascular bed. There was a significant interanimal variability. Thus, indomethacin may reduce the risk for PIVH by limiting cerebral blood flow, especially during increased cerebral perfusion pressures, which often occur after birth asphyxia. PMID- 8233723 TI - Escherichia coli 0111 B4 lipopolysaccharide given intracisternally induces blood brain barrier opening during experimental neonatal meningitis in piglets. AB - Neonatal bacterial meningitis remains a life-threatening infection, and severe neurologic sequelae may be left in survivors as well. The goal of the study was to develop and characterize a porcine model of the disease with intravital observation of the permeability changes in cerebral microvessels. Eighteen newborn piglets were given doses of 0 ng (group 1), 20 ng (group 2), and 200 ng (group 3) of Escherichia coli 0111 B4 endotoxin (LPS) intracisternally (n = 6 in each group). Cardiovascular parameters were without changes, but a compensated metabolic acidosis occurred in group 3 4 h after LPS injection. Using the open cranial window technique combined with fluorescence excitation, there was no blood-brain barrier leakage in pial-arachnoid microvessels for sodium fluorescein during the 4 h of experiments in group 1 piglets, whereas spotty extravasations occurred in group 2 and in group 3 after the LPS injections (70.5 +/- 10.5 and 55.2 +/- 4.1 min, respectively, mean +/- SEM). A dose-dependent increase in sodium fluorescein uptake in brain regions examined (parietal and occipital cortex, cerebellum, and periventricular white matter) was also found by fluorescence spectrophotometry. LPS-treated piglets had developed pleocytosis. Four h after the challenge, the white blood cell counts in cerebrospinal fluid were (mean +/- SD): group 1, 8.2 +/- 7.6 microL-1; group 2, 453 +/- 703 microL-1; and group 3, 1 027 +/- 620 microL-1, respectively, whereas there was no change in white blood cell count of peripheral blood samples.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233724 TI - Potentiation of N-methyl-D-aspartate-mediated brain injury by a human immunodeficiency virus-1-derived peptide in perinatal rodents. AB - In this study, we tested the hypothesis that human immunodeficiency virus (HIV)-1 derived peptides augment the neurotoxicity of excitatory amino acid agonists in vivo in postnatal day (PND) 7 rats. Stereotaxic intracerebral injections of the excitatory amino acid agonist N-methyl-D-aspartate (NMDA), alone or coinjected with an HIV-derived recombinant fusion peptide envelope gag (env-gag) were performed in PND-7 rats [group I: 5 nmol NMDA, n = 20; group II: 5 nmol NMDA + low-dose (1 or 50 ng) env-gag, n = 27; group III: 5 nmol NMDA + high-dose (100 ng) env-gag, n = 20], and brain injury was evaluated on PND 12. Based on histopathology scoring and measurements of hippocampal cross-sectional areas in the injected and contralateral hemispheres, coinjection of 100 ng of env-gag with 5 nmol of NMDA markedly increased the severity of resulting injury (p < 0.002, comparing histopathology scores; p < 0.003, comparing interhemispheric differences in hippocampal areas). These data suggest that in the developing nervous system HIV neurotoxicity may result, at least in part, from overactivation of excitatory amino acid receptors, and that perinatal rodent models may provide clinically relevant insights about the pathophysiology of HIV mediated brain injury. PMID- 8233725 TI - Complement activation in newborn infants with early onset infection. AB - The complement system is an important element in host defense. Quantitative deficiencies of total hemolytic complement activity and decreased C3 levels were reported in sera from normal neonates. However, little is known about complement activation products in the newborn. In a prospective study, complement activation products were determined in 32 healthy term neonates, in 41 neonates with colonization of their mothers, in 15 colonized neonates, and in 10 neonates with early onset infection. In all newborns, EDTA plasma was obtained within the first 6 h of life. The anaphylatoxin C3a-desArg was determined with a novel ELISA using an MAb reacting with a neoepitope of C3a-desArg. C3bBbP (alternative pathway convertase) and C1rsC1-inactivator (activation product of classical pathway) were measured with double-sandwich ELISA. C3 was determined by radial immunodiffusion. Plasma concentrations of C3a-desArg were similar in healthy term neonates and healthy adults, whereas diminished C3 levels were observed in the newborn infants. There were no significant differences between healthy neonates, neonates with colonized mothers, and colonized neonates. In neonates with infection, a significant elevation of C3a-desArg was found at the onset of the disease, resulting from alternative pathway activation. In contrast, the C1rsC1 inactivator complex showed no significant differences among healthy, colonized, and infected neonates. The anaphylatoxin C3a mediates inflammatory reactions such as vasodilatation and an increase in microvascular permeability and might therefore play an important role in severe neonatal infection. PMID- 8233726 TI - Human colostrum has anti-inflammatory activity in a rat subcutaneous air pouch model of inflammation. AB - An animal model was used to examine the effect of human colostrum on an acute inflammatory process in vivo. Subcutaneous air pouches on the backs of outbred rats were injected with carrageenan as an inflammatory challenge, normal saline, pooled aqueous human colostrum, or carrageenan plus colostrum concurrently. Oral dexamethasone or indomethacin was administered to some animals before and during challenge as anti-inflammatory agents. Polymorphonuclear leukocyte (PMN) counts in pouch fluid were determined 6 h postchallenge. Carrageenan challenge resulted in a significant acute inflammatory response [48.8 +/- 4.9 x 10(6) PMN/pouch (mean +/- SEM, n = 46)] compared with normal saline controls [0.9 +/- 0.2 x 10(6) (n = 31, p < 0.001 versus carrageenan)] or with colostrum [4.3 +/- 0.8 x 10(6) PMN/pouch (n = 11, p < 0.001 versus carrageenan)]. The concurrent injection of colostrum plus carrageenan challenge significantly reduced the PMN response compared with carrageenan alone [18.8 +/- 2.9 versus 48.8 +/- 4.9 x 10(6) PMN/pouch (carrageenan plus colostrum versus carrageenan, n = 41 versus 46, p < 0.001)]. This degree of suppression of PMN influx was not significantly different from that seen with indomethacin treatment but was significantly more than that seen with dexamethasone treatment. The decreases in PMN counts observed most likely reflect suppression of the acute inflammatory response because a significant amount of PMN lysis in colostrum was not observed in vitro and the accumulation of PMN granule contents was not seen in pouch fluid from colostrum treated animals in vivo. These data directly demonstrate for the first time that human colostrum has a biologically significant effect on the inflammatory process in vivo. PMID- 8233727 TI - Cytokine production in mononuclear cells of human milk studied at the single-cell level. AB - In this study, we demonstrate that mononuclear cells of human milk have a potential for production of many different cytokines. We applied a technique for cytokine detection at the single-cell level using cytokine specific MAb and immunofluorescence. The characteristic staining pattern obtained represents intracellular cytokine production, which allows for the assessment of the cellular origin of production. Milk mononuclear cells were mitogen-stimulated in vitro and cultured for 4 h and then stained for 13 cytokines. Lipopolysaccharide stimulation induced extensive production of the following monokines: IL-1 alpha, IL-1 beta, IL-1ra, IL-6, IL-8, and tumor necrosis factor-alpha. IL-10 and granulocyte-macrophage colony-stimulating factor were smaller products, although detectable in most samples. The abundant monokine production correlated with the high number of macrophages in milk. Spontaneous monokine production in unstimulated cells could be detected in six out of 11 samples. The highest incidence was evident for IL-8. No spontaneous lymphokine production was detected. Considering the low proportion of lymphocytes, stimulation with phorbol myristate acetate in combination with ionomycin resulted in considerable production of the following lymphokines: IL-2, IL-3, IL-4, IL-10, interferon gamma, tumor necrosis factor-alpha. Macrophages contributed to the high production of tumor necrosis factor-alpha and GM-CSF. IL-5 synthesis was detectable in only one sample. This work reveals that human milk mononuclear cells are potent producers of cytokines when mitogen stimulated in vitro. The in vivo implications of these findings remain to be investigated further. PMID- 8233728 TI - The effect of caloric supplementation on selected milk protective factors in undernourished Guatemalan mothers. AB - The level and avidity indices of specific antibodies against tetanus toxoid, Escherichia coli O6 and a pool of 10 common E. coli O antigens, as well as the concentration and daily output of lactoferrin and total secretory IgA (SIgA), were evaluated in the milk of moderately undernourished mothers who were in a random blind design divided into two groups and given different caloric supplementations. Group A received a high caloric supplement (500 kcal/d), and group B received a low caloric supplement (140 kcal/d). Determinations were done using ELISA in various modifications, except for lactoferrin, which was quantified by single radial immunodiffusion. The avidity indices were investigated as an evaluation of the antibody quality. In all the parameters evaluated, the only difference found between the two groups at the end of the supplementation period was in the content of total SIgA, which was lower in group B, both in concentration and daily output. However, the SIgA remained within the normal range. Increases as well as decreases in the levels of specific IgA antibodies occurred within both groups. Avidity was decreased in group B only against one of the antigens tested. We conclude that moderate undernutrition does not impair the levels of milk antibodies, and supplementation does not enhance them but prevents the decrease in the content of total milk SIgA. There is a suggestion that the avidity of certain antibody specificities could be hampered. PMID- 8233729 TI - Endogenous nitric oxide protects against platelet-activating factor-induced bowel injury in the rat. AB - Platelet-activating factor (PAF) causes bowel necrosis in animal models that is histologically identical to that seen in neonatal necrotizing enterocolitis, but little is known about endogenous mechanisms that might protect against PAF induced bowel injury. We hypothesized that endogenous nitric oxide might represent such a protective mechanism. Adult male Sprague-Dawley rats were pretreated with 2.5 mg/kg NG-nitro-L-arginine methyl ester (L-NAME), a potent nitric oxide synthase inhibitor, and given injections of 1.5 micrograms/kg PAF 15 min later. Animals treated with normal saline placebo, L-NAME alone, and PAF alone were also studied. Superior mesenteric artery blood flow and blood pressure were continuously recorded. At the end of 2 h or upon death of the animal, hematocrit was measured and intestinal samples were taken for histologic examination and determination of myeloperoxidase activity, a measure of intestinal neutrophil content. Compared with animals given PAF alone, animals pretreated with L-NAME followed by PAF developed significantly worse bowel injury (median injury scores: 2.5 versus 0.5, p = 0.005), hemoconcentration (final hematocrit 65.2 +/- 2.0% versus 53.9 +/- 1.0%, p < 0.001), and intestinal myeloperoxidase activity (12.45 +/- 1.94 U/g versus 6.51 +/- 0.57 U/g, p < 0.01). The last two effects were further accentuated when 10 mg/kg L-NAME was given before PAF. Treatment with sodium nitroprusside, a nitric oxide donor, for 10 min before and after PAF administration reversed the effects of L-NAME. Animals pretreated with phenylephrine rather than L-NAME did not develop worse injury than animals treated with PAF alone despite comparable reductions in superior mesenteric blood flow before PAF treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233730 TI - Spontaneous gonadotropin and testosterone concentration profiles in prepubertal and pubertal boys: temporal relationship between luteinizing hormone and testosterone. AB - To investigate the detailed pattern of change in circulating gonadotropin and testosterone concentrations around the onset of puberty and to determine the temporal relationship between the gonadotropin and testosterone secretion, plasma gonadotropin and testosterone were measured at 20-min intervals for 24 h in 21 normal short boys. The obtained plasma hormone concentrations were analyzed by Cluster pulse detection algorithm, cosinor analysis, and cross-correlation analysis. The 21 subjects were divided into the prepubertal (n = 16) and early pubertal (n = 5) groups. All subjects showed nocturnal LH and FSH pulses and had significant circadian LH and FSH rhythms. Except for six boys of prepubertal group, all subjects showed nocturnal testosterone pulses and had significant circadian testosterone rhythms. The acrophase (clocktime of maximal value) of circadian testosterone rhythm was 0308-0428 h. Cross-correlation analysis demonstrated significant positive cross-correlations between LH and testosterone that were maximum at a testosterone lag of 60-120 min. Further, to eliminate intrinsic autocorrelations within the LH and testosterone time series, we filtered the data before subjecting them to the cross-correlation analysis. As a result, significant positive cross-correlations were found at a testosterone lag of 40 min in 10 peripubertal boys. We conclude that testosterone concentration profiles are pulsatile and show marked circadian rhythm well before the onset of puberty. LH and testosterone time series are significantly coupled when testosterone lags LH by about 40 min. This time lag might correspond to the time for synthesizing and secreting testosterone in Leydig cells after binding of LH to the Leydig cell receptors. PMID- 8233731 TI - Platelet-activating factor-induced ischemic bowel necrosis: the effect of platelet-activating factor acetylhydrolase. AB - In the present investigation, the rat model of necrotizing enterocolitis (NEC) was further developed by injection of platelet-activating factor (PAF) in the descending aorta. The role of the plasma PAF-acetylhydrolase (PAF-AH) was examined in the prevention of this disease. PAF (0.35 micrograms) caused ischemic intestinal necrosis when administered intraaortically. The effects of PAF injection on the small intestine were examined histologically in samples of the duodenum, jejunum, and ileum. The administration of PAF resulted in extensive hemorrhagic damage in all regions of the small bowel and a marked hemoconcentration. Pretreatment of the rats with dexamethasone or medroxyprogesterone significantly increased plasma PAF-AH activity. Dexamethasone and medroxyprogesterone prevented the gross and histologic features of NEC as well as the hemoconcentration. In contrast, lower amounts of PAF were sufficient to cause bowel necrosis and hemoconcentration when decreased activities of plasma PAF-AH were induced by 17 alpha-ethynylestradiol or 4-aminopyrazolopyrimidine administration. We have recently reported that PAF-AH is present in human milk. The beneficial effect of breast feeding in preventing the development of NEC in the newborn is discussed and a mechanism proposed to explain this finding. It is suggested that PAF may play an important role in the pathogenesis of NEC and that the increased plasma activity of PAF-AH caused by dexamethasone and the presence of this enzyme, of milk origin, in the lumen of the small bowel may prove to be beneficial in the prevention of this disease. PMID- 8233732 TI - Annual summary of vital statistics--1992. AB - A new low in the infant mortality rate was reached in 1992, at 848.7 deaths per 100,000 live births, a decline of 5% from 894.4 in 1991. Birth, death, and marriage rates were also lower, but the divorce rate inched up to 4.8 per 1000 population, the same level as in 1988. The age-adjusted death rate was 504.9 per 100,000 population, the lowest in US history. Natural increase in the population, excess of births over deaths, decreased from 1,941,389 to 1,907,000, from 7.7 to 7.5 per 1000 population. Births outside hospital were fewer, both in numbers and in proportion to all births. Birth rates increased at both ends of the age range but declined in the principal childbearing years. Births to unmarried mothers increased again, comprising more than one fifth of white births and two thirds of black births. A higher proportion of newborns weighed less than 2500 g than in 1989. Life expectancy at birth increased again, to 75.7 years overall, paralleled in both sexes and white and black races. The age-adjusted death rate for cardiovascular diseases declined, but malignancies of the respiratory system increased again, to almost six times what it was in 1940. Chronic obstructive pulmonary diseases, despite slight improvement since 1991, caused death more than eight times as often as in 1940. Black and white infant mortality rates both showed a decline, greater in the white neonatal component; the black/white discrepancy widened slightly. Infant mortality in those of Hispanic origin was slightly higher than non-Hispanic whites, but the National Center for Health Statistics warns that Hispanic rates may be understated. There was little change in causes of infant mortality, or in black to white ratios for the several causes. On the world scene, most industrialized countries showed declines in infant mortality matching the US. In 1991, 21 countries, 15 in Europe, 3 in Asia, 2 in Oceania, and 1 in North America, had infant mortality rates less than the US. The decline in most other countries has been more rapid than in the US. PMID- 8233733 TI - Randomized trial of silver nitrate, erythromycin, and no eye prophylaxis for the prevention of conjunctivitis among newborns not at risk for gonococcal ophthalmitis. Eye Prophylaxis Study Group. AB - OBJECTIVE: To compare the efficacy of commonly used forms of eye prophylaxis for newborns with no prophylaxis in the prevention of nongonococcal conjunctivitis. DESIGN: Randomized doubly masked clinical trial. SETTING: University of Washington Hospital and affiliated clinics, Seattle, between 1985 and 1990. SUBJECTS: The medical records of 8499 women were evaluated for possible participation; 2577 were eligible. Of the 758 enrolled, the infants of 630 were evaluable. INTERVENTION: Comparison of silver nitrate, erythromycin, and no eye prophylaxis given at birth for the prevention of conjunctivitis. MAIN OUTCOME MEASURES: Conjunctivitis during the first 60 days of life and nasolacrimal duct patency in the first 2 days of life. RESULTS: The frequency of impatent tear ducts at the 30- to 48-hour examination did not differ significantly by prophylaxis group. Among the 630 infants randomized and observed, 109 (17%) developed mild conjunctivitis. Sixty-nine (63%) of the cases appeared during the first 2 weeks of life. After 2 months of observation, infants allocated to silver nitrate eye prophylaxis at birth had a 39% lower rate of conjunctivitis (hazard ratio = 0.61, 95% confidence interval = 0.39 to 0.97), and those allocated to erythromycin had a 31% lower rate of conjunctivitis (hazard ratio = 0.69, 95% confidence interval = 0.44 to 1.07), than did those allocated to no prophylaxis. CONCLUSION: Silver nitrate eye prophylaxis caused no sustained deleterious effects and even provided some benefit to infants born to women without Neisseria gonorrhoeae. However, the effect was modest and against microorganisms of low virulence. The results suggest that parental choice of a prophylaxis agent including no prophylaxis is reasonable for women receiving prenatal care and who are screened for sexually transmitted diseases during pregnancy. PMID- 8233734 TI - Penicillin-resistant systemic pneumococcal infections in children: a retrospective case-control study. AB - OBJECTIVE: To determine whether there are any risk factors that differentiate children with systemic infections due to Streptococcus pneumoniae relatively or fully resistant (minimum inhibitory concentration > 0.1 microgram/mL) to penicillin from those children with infections due to S pneumoniae susceptible to penicillin. DESIGN: Retrospective case-control study. SETTING: A large children's hospital. PARTICIPANTS: Forty-three children with systemic penicillin-resistant S pneumoniae infections identified at Texas Children's Hospital over the 51-month period from January 1989 through March 1993. Each case had one or two controls matched only for age and date of S. pneumoniae infection. Sixty-six controls were selected from a group of 341 children with susceptible isolates. OUTCOME MEASURES: Variables compared included gender, race, diagnosis, underlying conditions, past hospitalization, geographic area of residence, antibiotic use in past month, amoxicillin-clavulanic acid use in past month, and outcome. RESULTS: Thirty-seven patients (86%) had relatively resistant isolates (minimum inhibitory concentration range 0.125 to 1.0 microgram/mL) and six patients (15%) had fully resistant isolates (minimum inhibitory concentration range 2.0 to 8.0 micrograms/mL). Thirty-three percent of the cases vs 36% of the controls had underlying conditions. Seventy-one percent of the cases vs 39% of the controls had received antibiotics in the previous month. Compared with their matched controls, the patients with penicillin-resistant systemic pneumococcal infections were more likely (P = .02) to have received a course of antibiotics within the month prior to their infection. CONCLUSION: The only identified associated risk factor in children who developed a systemic penicillin-resistant pneumococcal infection appears to have been the use of antibiotics within the month prior to their infection. PMID- 8233735 TI - Comparison of prophylaxis and rescue treatment with Curosurf in neonates less than 30 weeks' gestation: a randomized trial. AB - OBJECTIVE: The aim of this randomized clinical trial was to evaluate the immediate effects of prophylactic administration of Curosurf and to compare outcomes after prophylactic or expectant management. STUDY DESIGN: Porcine surfactant (Curosurf, 200 mg/kg body weight) was administered intratracheally within 10 minutes of birth to preterm neonates with a gestational age of 26 to 29 weeks (n = 75); rescue-eligible neonates (n = 72) were initially subjected to a sham maneuver. The primary end points of the trial, evaluated at the age of 6 hours, were to obtain (1) a 40% decrease in the ratio between transcutaneous oxygen tension (tcPO2) (kPa) and fraction of inspired oxygen (FIO2), and (2) a 50% decrease in the incidence of radiologically verified respiratory distress syndrome (RDS). After 6 to 24 hours, a similar dose of surfactant was given to the neonates of both the prophylaxis and the rescue-eligible group, if they needed mechanical ventilation with an FIO2 > or = 0.6. RESULTS: At 6 hours the prophylaxis group had, in comparison with the rescue-eligible group, significantly higher tcPO2/FIO2 ratios (mean +/- SD: 39.7 +/- 15.3 vs 28.1 +/- 18.1; P < .001) and less severe RDS by radiological scoring (chi 2 = 14.9; P = .005). Severe RDS was present in 19% of the prophylactically treated neonates versus 32% in the rescue-eligible group (P < .05). The prophylaxis group needed shorter periods of FIO2 > 0.40 than the rescue-eligible neonates (P < .01), and eight neonates of the prophylaxis group (11%) versus 23 of the rescue-eligible group (32%) qualified for rescue treatment with surfactant in the interval 6 to 24 hours (P < .01). There were no differences in the incidence or severity of pneumothorax, pulmonary interstitial emphysema, cerebral hemorrhage, periventricular leukomalacia, patent ductus arteriosus, in the duration of mechanical ventilation or time in supplemental oxygen, or in mortality. CONCLUSIONS: Subgroup analysis revealed (1) that administration of corticosteroids reduced the risk of developing neonatal RDS as effectively as did surfactant prophylaxis at birth, and (2) that prophylaxis was effective especially in neonates with gestational age < 28 weeks or birth weight < 1000 g, in male neonates, and in neonates who had received no antenatal treatment with corticosteroids. Our data indicate that prophylactic treatment with surfactant should be considered in high-risk neonates fulfilling these latter criteria. PMID- 8233736 TI - Surfactant therapy and intracranial hemorrhage: review of the literature and results of new analyses. AB - BACKGROUND AND OBJECTIVE: Surfactant replacement is a powerful therapy for newborns with respiratory distress syndrome, but limited observations suggest that alterations of cerebral blood flow can accompany the use of several available surfactants. An early European multicenter controlled study with beractant demonstrated an increased rate of intracranial hemorrhage in treated patients. Nine additional controlled studies were subsequently performed and included follow-up evaluations through 2 years adjusted age. This clinical experience provided a database of approximately 1700 infants to examine retrospectively for any relationship between surfactant therapy and intracranial hemorrhage. METHODS: Cumulative incidence rates, hazard ratios, and 95% confidence intervals for intracranial hemorrhage were computed for each study and for appropriately pooled studies of similar design. Where an association between surfactant and the risk of intracranial hemorrhage was found, additional analyses were performed to attempt to identify intermediate physiologic events that might link administration of surfactant to the occurrence of intracranial hemorrhage. These analyses were guided by literature reports of hemodynamic changes observed in association with surfactant therapy. RESULTS: During the controlled studies with beractant, treated newborns of 600 to 750 g birth weight were at higher risk for grades I and II intracranial hemorrhage than control newborns. There was no increased risk for grades III and IV hemorrhage among these newborns, nor was there increased risk of hemorrhage among any other patient groups. This finding did not result in increased morbidity for the affected patients; at 2 years adjusted age, they were not different from the control infants of 600 to 750 g birth weight. Retrospective examination of the database could not pinpoint the mechanism behind the finding, but it might have been related to changes in cerebral blood flow after surfactant uncompensated by ventilator management of oxygenation and ventilation. CONCLUSIONS: Surfactant therapy may set in motion hemodynamic changes that could predispose to intracranial hemorrhage in certain circumstances, but this can probably be compensated by careful management of oxygenation and ventilation. A relationship between surfactant therapy and intracranial hemorrhage is probably not isolated to any particular surfactant preparation or method of delivery; studies comparing surfactants have shown no differences in rates of intracranial hemorrhage. PMID- 8233737 TI - Ocular sequelae in premature infants. AB - OBJECTIVE: The authors report on the incidence of myopia and strabismus at 12 and 24 months postterm in a cohort of 190 premature infants with birth weights of less than 1251 g born in 1986 and 1987. METHODS: The neonatal and follow-up eye charts of a cohort of 190 premature infants were retrospectively reviewed. All 138 children who survived the neonatal period had at least one eye examination between day 28 and 42 of life that documented the presence and staging of retinopathy of prematurity (ROP) according to the International Classification of ROP. No infants received cryotherapy. Eye examinations conducted at 12 and 24 months postterm included assessment of vision, fundus, ocular motility, anterior segment abnormality, and refractive error. Eyes were refracted using cycloplegic retinoscopy. Strabismus was detected using the Hirschberg and cover tests. Eye reports were available for 80% (n = 110) at 12 months and 36% (n = 50) at 24 months. RESULTS: Fifty-three percent of the cohort exhibited ROP in the neonatal period; 12% of these progressed to stage 3 or 4 ROP. Myopia was observed in 16% (18/110) of the cohort at 12 months of age; 4.5% (5/110) measured more than 4.0 diopters of myopia. Children with birth weights of less than 751 g were 3.2 times more likely than those with birth weights between 751 and 1000 g and 10 times more likely than those with birth weights between 1001 and 1250 g to develop myopia in the first year of life. The likelihood of myopia at 12 months doubled with each increment in ROP stage. Of the 50 children reexamined at 24 months postterm, more than 80% demonstrated deteriorating vision. The incidence of myopia increased to 38% (19/50) overall, with 24% (12/50) of the cohort showing severe myopia. Astigmatism and anisometropia were highly correlated with severe myopia. Strabismus was seen with increasing frequency through the second year of life. All children with grade III or IV intraventricular hemorrhage in the neonatal period developed esotropia. CONCLUSION: This study emphasizes the significant roles of low birth weight, ROP, and intraventricular hemorrhage in the development of myopia and strabismus. Follow-up to 2 years of life is recommended given the demonstrated deterioration in our cohort. PMID- 8233738 TI - Circumcision in children beyond the neonatal period. AB - OBJECTIVE: Although many boys are circumcised after the newborn period, there are limited data concerning the proportion of uncircumcised infants who subsequently have the procedure, as well as minimal information about characteristics of this population. POPULATION: Data and medical records of boys born in US Army hospitals from 1985 through 1992 were reviewed. General information regarding a portion of the 32,072 boys who were not circumcised during their neonatal hospitalization, but subsequently had the procedure, is reported. In addition, specific details are presented about 476 boys who were circumcised after their birth hospitalization. RESULTS: The postneonatal circumcision frequency rate of these children increased significantly over this period (P < .0001), particularly after the 1989 American Academy of Pediatrics statement on circumcision. In general, most boys were not circumcised as neonates because of parental choice (55%) or because of a complicated neonatal course (32%). "Sleeve" and "freehand" resection were the most common methods of circumcision (93%), and general anesthesia was used in 91% of the boys. The mean age at postneonatal circumcision was 2.92 years. Common reasons for the ensuing procedure included parental choice (39%), coincidence with other surgery (27%), recurrent balanoposthitis (23%), and urinary tract infections (7%). Complications occurred in 8 of the 476 boys whose specific records were reviewed. These included excessive bleeding (3), malignant hyperthermia (2), aspiration pneumonia (1), large hematoma development (1), and postoperative fever (1). CONCLUSIONS: An increasing number of boys are circumcised beyond the neonatal period, and the procedure performed at this time can be associated with substantial complications. This information may be of use when performing informed-consent counseling for neonatal circumcision. PMID- 8233739 TI - The inconspicuous penis. AB - OBJECTIVE: To describe the etiology and management of the group of abnormalities referred to as the inconspicuous penis. DESIGN: Analysis of 19 cases seen over a period of 2 years by chart review. SETTING: Children's hospital in a major metropolitan area. PATIENTS: Nineteen boys referred to two pediatric urologists over a period of 2 years with penises that appeared abnormally small, but on palpation and measurement, were found to have a normal shaft with a normal stretched length. Diagnoses included were buried penis, webbed penis, and trapped penis. Patients ages ranged from 1 week to 13 years. FINDINGS: There were eight patients (42%) with trapped penis, and all were complications of circumcision (age 1 week to 7 months). Of nine (47%) patients with buried penis, two had been circumcised prior to diagnosis. One (5%) patient had webbed penis and one (5%) had combined buried and webbed penis. INTERVENTION: Six trapped penises were surgically repaired, and two resolved spontaneously. Five patients with buried penis had surgical repair, and two are being followed up for probable repair at age 9 to 12 months. Two were not repaired because of medical conditions or parental concerns. The webbed penis was surgically repaired as was the combined buried and webbed penis. The repair were all successful and had no complications. CONCLUSIONS: Inconspicuous penis encompasses a group of conditions in which the penis appears small but the shaft can be normal or abnormal in size. Circumcision is contraindicated in these patients until they have been evaluated by a urologist. Further study is needed to determine the natural history of these disorders and to better define which patients will benefit from surgical intervention and at what age. PMID- 8233740 TI - Clinical features and epidemiology of invasive Kingella kingae infections in southern Israel. AB - OBJECTIVE: To characterize the clinical spectrum and epidemiology of invasive Kingella kingae infections in children living in southern Israel. DESIGN: Five year observational, descriptive study. POPULATION: Children in whom K. kingae was isolated from blood or other normally sterile body fluid. RESULTS: Twenty-five patients with invasive K. kingae infection (13 male and 12 female) were identified. Twenty-four of these children were younger than 2 years. The annual incidence was 14.3, 27.4, and 31.9 cases per 100,000 children < or = 4 years, < or = 24 months, and < or = 12 months, respectively. Seventeen (68%) of 25 patients sought treatment between July and December. Concomitant upper respiratory tract infection or stomatitis was observed in 14 (56%) of the patients, suggesting a respiratory or buccal source for the infection. Four children were bacteremic: 2 of them suffered from a lower respiratory tract infection, and the remaining 2 had bacteremia with no evident focal infection. Twenty-one children had skeletal infections and none of them was bacteremic; 16 had septic arthritis, 3 had osteomyelitis, 1 had both osteomyelitis and septic arthritis of the adjacent joint, and 1 had dactylitis of the hand. Involvement of the ankle was unusually frequent among children with septic arthritis, whereas the calcaneus was involved in 3 of the 4 children with osteomyelitis. Antibiotic treatment resulted in full recovery in all cases, and only 2 patients with septic arthritis required surgical drainage. CONCLUSION: Kingella kingae is a much more common cause of invasive infection in young children than has been previously recognized. The disease has a clear seasonal pattern, usually affects the skeletal system, frequently involves unusual bones and joints, and follows a benign course. PMID- 8233741 TI - Adolescent gestational weight gain and birth weight. AB - OBJECTIVE: To examine the relationship among maternal age, prepregnancy weight, gestational weight gain, and birth weight in 141 low-income black adolescents and their infants. STUDY SAMPLE: One hundred forty-one consecutively enrolled, low income, black adolescents who entered prenatal care prior to their 23rd week of gestation, were free of chronic diseases, took no regular medications, had no known uterine anomalies, and gave birth to one live neonate. RESULTS: After controlling for prepregnancy weight and other potentially confounding variables, we found a significant relationship between gestational weight gain and infant birth weight among younger adolescents (< 16 years old at conception), but not among older adolescents (16 through 19 years old at conception); younger adolescents contributed more of their gestational weight gain to their fetuses than did older adolescents. Among younger adolescents the rate of maternal weight gain during the entire gestation was significantly correlated with birth weight (r = .40; P < .01), whereas for older adolescents only maternal weight gain during the second half of gestation was significantly correlated with birth weight (r = .25, P < .05). CONCLUSIONS: The data do not support the thesis that younger/adolescents compete with their fetuses for nutrients; in fact, younger study adolescents transferred more of their gestational weight gain to their fetuses than did older adolescents. PMID- 8233742 TI - Apnea and factitious illness (Munchausen syndrome) by proxy. AB - OBJECTIVE: Munchausen syndrome by proxy (MSP) is recognized in the differential diagnosis of apparent life-threatening events, but the early signs and the full spectrum of this presentation are not well recognized. We aim to describe MSP presenting with apnea to illustrate this spectrum and the evolution in our management over a period of 10 years. PATIENTS AND RESULTS: Eleven children in five families seen in one institution and assessed by one team are described in detail. The children had apnea and/or pallor, but with a wider age range than usually seen with apparent life-threatening events, sometimes associated with other injuries, and a large percentage of parents were health care providers. In no case was apnea witnessed by health care professionals other than the parents. There were frequent disagreements in management between professionals and, consequently, delays in considering the diagnosis at first. There were two deaths. A team developed, allowing the diagnosis of MSP to be considered sooner and the cases to be assessed and managed consistently. CONCLUSION: MSP is part of child abuse, and it needs to be recognized by all physicians. Family assessment is required and the development of a team interested in MSP facilitates assessment and management. PMID- 8233743 TI - Maternal smoking before and after pregnancy: effects on behavioral outcomes in middle childhood. AB - OBJECTIVE: The aim of this research was to examine the extent to which maternal smoking before and after pregnancy was associated with childhood disruptive behaviors when due allowance was made for potentially confounding factors which may have been associated with both maternal smoking habits and childhood problem behaviors. METHODS: During the course of a 15-year longitudinal study of a birth cohort of 1265 New Zealand children, the following measures were obtained: (1) measures of daily cigarette intake during pregnancy and after pregnancy; (2) measures of childhood disruptive behaviors including conduct problems and attention deficit behaviors based on both maternal and teacher report data; and (3) measures of a series of potentially confounding family, social, parental, and related factors. RESULTS: Before adjustments for confounding, maternal smoking both before and after pregnancy was found to be associated with significant increases in rates of childhood problem behaviors: children whose mothers smoked in excess of 20 cigarettes per day had mean problem behavior scores that were between 0.16 and 0.56 standard deviations higher than those of children whose mothers were nonsmokers. The results were then adjusted using regression methods to take account of (1) correlations between pregnancy and postpregnancy smoking and (2) potentially confounding factors. The results of regression adjustment suggested that maternal smoking during pregnancy was associated with small but statistically detectable increases in rates of childhood problem behaviors, with children whose mothers smoked in excess of 20 cigarettes per day having mean scores that were 0.10 to 0.36 standard deviations higher than those of the offspring of nonsmokers, even after adjustment for a series of confounding factors. However, smoking after pregnancy was not significantly associated with increased rates of childhood problem behavior after adjustment for sources of confounding. CONCLUSION: The results are generally consistent with the hypothesis that smoking during pregnancy may be associated with small but detectable increases in the risks of problem behaviors in childhood. This suggests that possible adverse effects on childhood behavioral adjustment should be added to the growing list of adverse consequences of parental smoking for childhood health and well-being. PMID- 8233744 TI - Left-handedness as a risk factor for unintentional injury in children. AB - OBJECTIVE: To determine whether left-handedness is a risk factor for unintentional injury among children and adolescents. DESIGN: Case-control study. SETTING: Pediatric emergency department of Arkansas Children's Hospital. PATIENTS: 265 patients sustaining unintentional trauma aged 6 to 18 years and 494 control patients who did not have trauma were given a questionnaire to determine handedness, past unintentional injury, and parental perception of injury proneness. RESULTS: The frequency of left-handedness in the trauma group (18.1%) was significantly greater than frequency of 10.5% in the control group (P < .003, odds ratio = 1.80, 95% confidence interval 1.20 to 2.72). Multivariate analysis revealed handedness as the only significant variable between trauma and control (P < .04). The proportion of left-handers who had been hospitalized previously for injury treatment (20.0%) was larger than the proportion of right-handers, (12.0%) (P < .026, odds ratio = 1.84, 95% confidence interval 1.03 to 3.27). More parents of left-handers rated their child as "more clumsy than average" than parents of right-handers (26.0% vs 15.2%, P < .007). CONCLUSIONS: Left-handedness appears to be a risk factor for unintentional injury in children and adolescents in a pediatric emergency department population. PMID- 8233745 TI - Safety and immunogenicity of a combined diphtheria, tetanus, pertussis and Haemophilus influenzae type b vaccine in young infants. AB - OBJECTIVE: To study the safety and immunogenicity of a combined diphtheria tetanus-pertussis (DTP)-Haemophilus influenzae type b (HbOC) vaccine (TETRAMUNE) in infants as young as 2 months of age as compared to separate administration of DTP and HbOC. METHODS: Two-month-old infants were randomized to receive three doses 2 months apart of either DTP-HbOC as a single 0.5-mL injection or to receive 0.5 mL of DTP and HbOC concurrently in separate legs. Local and systemic adverse reactions were monitored within 72 hours of each immunization, and immunogenicity of each of the four vaccine components was measured. RESULTS: The incidence of both local and systemic adverse events following the tetravalent vaccine was similar to the incidence following separate vaccine administration. After three doses of vaccine, the response to each of the vaccine components was higher in the combined vaccine when compared to separate administration. In the case of the Haemophilus influenzae type b component, this enhancement was also seen after two doses. The response to the combined vaccine was consistent among the three lots tested as was the enhancement over separate administration. CONCLUSIONS: The DTP-HbOC vaccine was safe and immunogenic in young infants and was generally more immunogenic than separate vaccination with DTP and HbOC. The use of such a combined vaccine reduces the number of injections given to young infants by half and is an important step toward improving vaccine delivery. PMID- 8233746 TI - Clinical survey of natural varicella compared with breakthrough varicella after immunization with live attenuated Oka/Merck varicella vaccine. AB - OBJECTIVE: To prospectively characterize varicella occurring in children previously immunized with a live attenuated varicella vaccine (breakthrough varicella) through daily observation by medical personnel and to compare it with natural varicella followed in the same manner. DESIGN: A blinded clinical survey. SETTING: Four pediatric practices (two private; two hospital-based). PARTICIPANTS: Healthy 12-month-old through 17-year-old children with chickenpox were studied; 92 had natural varicella and 58 had breakthrough varicella. SELECTION PROCEDURES AND INTERVENTIONS: Study personnel, unaware of vaccination status, documented the clinical characteristics of each patient in the office or at the patient's home each day from enrollment until the day after the total number of lesions increased less than 10%. A standard form documenting number and description of lesions, temperature, duration of illness, and associated clinical complaints was completed each day by the same study personnel. Acute and convalescent sera were obtained on breakthrough cases. MEASUREMENTS AND RESULTS: Antibody to varicella-zoster virus was measured by the glycoprotein-based enzyme linked immunosorbent assay. Of those with sera available, 85% were serologically confirmed. Eighty-seven percent of enrollees had a known exposure to chickenpox, with at least two thirds of each group having a greater than 4-hour or a household exposure. The numbers of total and vesicular lesions were significantly higher in the natural varicella group, regardless of exposure status (P = .021 to < .001). The group with breakthrough varicella had a significantly lower incidence of fever (P < .001) and a significantly shorter duration of illness (P < .001). Other associated constitutional complaints and complications were not significantly different between groups. CONCLUSION: Varicella in vaccine recipients is clinically modified and significantly less severe than natural disease. PMID- 8233747 TI - Neurodevelopmental testing of children born to human immunodeficiency virus type 1 seropositive and seronegative mothers: a prospective cohort study in Kigali, Rwanda. AB - OBJECTIVE: The results of developmental testing of 218 children born to human immunodeficiency virus (HIV)-seropositive mothers and infected or uninfected themselves were compared with those of 218 children born to HIV-seronegative mothers in an ongoing cohort study in Kigali, Rwanda. METHODS: When the children were 6, 12, 18, and 24 months of age, a specific neurodevelopmental examination was performed blindly by study physicians assessing gross motor development, fine motor development, language acquisition, and social contacts. RESULTS: Only one acute severe HIV-related encephalopathy was identified among the 50 infected children. The proportion of abnormal neurologic examinations in HIV-infected children varied from 15% to 40% according to age and was always higher than in HIV-uninfected children born to HIV-seropositive and seronegative mothers (< or = 5% or less of abnormal examinations at each time period). After excluding those children with clinical acquired immunodeficiency syndrome (AIDS) from the analysis, the proportion of abnormal examinations in infected children was 12.5% at 6 months, 16% at 12 months, 20% at 18 months, and 9% at 24 months of age and was still more frequent than in HIV-uninfected children. The developmental delay was principally due to significantly lower gross motor scores. CONCLUSIONS: HIV-1 infected children are more frequently developmentally delayed than uninfected children during the first 2 years of life in this African population. This developmental delay is related to the AIDS stage of pediatric HIV infection. PMID- 8233748 TI - Childhood-onset systemic lupus erythematosus: antiphospholipid antibodies in 37 patients and their first-degree relatives. AB - OBJECTIVE: Antiphospholipid antibodies (aPL) are noted with increased frequency in patients with systemic lupus erythematosus (SLE). The main manifestations found to be associated with aPL are arterial and venous thrombotic events, thrombocytopenia, and recurrent pregnancy loss. This study is an attempt to define the incidence of aPL in patients with childhood-onset SLE and in their relatives and to correlate their presence with clinical manifestations, and especially, to evaluate the risk of thrombosis in aPL-positive subjects. METHODOLOGY: We studied 37 unrelated patients and 107 of their first-degree relatives. VDRL, IgG and IgM anticardiolipin, and IgG antiphosphatidylethanolamine antibodies were studied in all probands during periods of clinical remission and in first-degree relatives at the time of interview. Lupus anticoagulant had only been studied in probands during an SLE flare-up. RESULTS: Thirty-eight percent of probands and 19% of relatives were positive for at least one aPL, with little overlap between the different aPL studied. -No aPL-negative proband developed thrombosis. Two of the aPL-positive probands had thrombotic events before testing, and a third one showed thrombosis after testing. Only two probands had high levels of IgG aCL and showed thrombosis. The occurrence of aPL positivity in relatives was not always related to its presence in probands. None of the aPL-positive relatives had had thrombosis, but recurrent fetal loss was noted in one aPL-positive mother with SLE. Although there was a high frequency of SLE, SLE-like disease, auto-immune disorders or positive serological findings for lupus in first-degree relatives, many of these relatives did not test positive for aPL. CONCLUSION: The high levels of IgG aCL may be considered a risk factor for thrombosis. Findings in relatives suggest a multifactorial origin for autoimmune disease and antibody production. PMID- 8233749 TI - Twinning and cerebral palsy: experience in four northern California counties, births 1983 through 1985. AB - BACKGROUND: Twinning is associated with heightened risk of cerebral palsy (CP) and is increasing in the United States and elsewhere. METHODS: Twins with moderate or severe congenital CP were identified in a cohort of 155,572 children born 1983 through 1985 in four northern California counties and surviving to 3 years. The prevalence of CP in twins and factors associated with increase in risk were examined. RESULTS: Among 2985 twins, 20 children in 18 pairs had CP. The prevalence of CP was 6.7 per thousand 3-year-old twin children (95% confidence interval [CI], 4.2 to 11), 12 per thousand twin pregnancies (95% CI, 7.2 to 19), and 1.1 per thousand singletons (95% CI, 0.97 to 1.3). Ten percent of all CP was in twins; 22% of CP in infants of less than 1500 g birth weight occurred in twins. Twins were over-represented among very low birth weight infants but their risk of CP was comparable with that of very low birth weight singletons. Twins born weighing 2500 g and more had a CP risk 3.6 times that of singletons of similar weight. In children who survived fetal death of a co-twin, CP was 108 times more prevalent (95% CI, 42 to 273) than in singletons and 13 times more prevalent (95% CI, 4.5 to 37) than in twins whose co-twin was born alive. The CP rate in unlike-sex pairs was 13 per thousand (95% CI, 4.8 to 32), not significantly different from 11 per thousand (95% CI, 5.7 to 19) for like-sex pairs. CONCLUSION: Twin pregnancies produced a child with CP 12 times more often than singleton pregnancies. The heightened risk was largely related to the tendency of twins to be low in birth weight and to a greater risk of CP in twins of normal birth weight compared with singletons of similar weight. Twins of unlike-sex pairs, necessarily dizygotic, were not at lower risk than like-sex pairs. The current increase in multiple births is likely to contribute more children with CP. PMID- 8233750 TI - The new National Institutes of Health strategic plan. PMID- 8233751 TI - Circumcision updated--indicated? PMID- 8233752 TI - Hypervitaminosis D after prolonged feeding with a premature formula. PMID- 8233753 TI - Papular-purpuric gloves and socks syndrome: a characteristic viral exanthem. PMID- 8233754 TI - Henoch-Schonlein purpura in infants. PMID- 8233755 TI - The origin of the National Institute of Child Health and Human Development. PMID- 8233756 TI - American Academy of Pediatrics Committee on Child Abuse and Neglect: Shaken baby syndrome: inflicted cerebral trauma. AB - The shaken baby syndrome is a clearly definable medical condition. It requires integration of specific clinical management and community intervention in an interdisciplinary fashion. PMID- 8233757 TI - American Academy of Pediatrics Committee on Children With Disabilities and Committee on Psychosocial Aspects of Child and Family Health: Psychosocial risks of chronic health conditions in childhood and adolescence. PMID- 8233758 TI - American Academy of Pediatrics Committee on Children With Disabilities: Provision of related services for children with chronic disabilities. PMID- 8233759 TI - Statistics and Swedish 'sambos'. PMID- 8233760 TI - What sunscreen should I use for my 3-month-old baby? PMID- 8233761 TI - Bike dangers. PMID- 8233762 TI - Near infrared spectroscopy research. PMID- 8233763 TI - Sleep, feeding, and opinions. PMID- 8233764 TI - Think globally; act locally. PMID- 8233765 TI - A comparison of paraplegic gait performance using two types of reciprocating gait orthoses. AB - This study examined the energy cost of ambulation using the reciprocating gait orthosis (RGO) and the modified Isocentric RGO in paraplegic spinal cord injured subjects. In 4 subjects, the rates of O2 consumption per minute, O2 cost per metre, heart rate (HR), respiratory exchange ratio, velocity, and physiologic cost index (PCI) were measured during ambulation with the two orthotic devices. PCI was calculated by dividing the difference between walking and resting HR by velocity. PCI was significantly lower during ambulation trials with the Isocentric RGO compared to the RGO, but was the only measurement that detected a significant difference between the two orthotic devices. These results indicate that energy costs of ambulation at self-selected speeds were lower with the Isocentric RGO compared to the standard RGO. Furthermore, PCI could be used as a sensitive indicator of gait efficiency in spinal cord injury subjects. PMID- 8233766 TI - Enhancement of hemiplegic patient rehabilitation by means of functional electrical stimulation. AB - This presentation will review briefly the current practice and state of the art in functional electrical stimulation (FES) as applied to stroke, head injured or brain tumour operated patients. A similar application is used in paretic patients following trauma or other aetiology. Over 20 years experience in the application of FES, as practised in Ljubljana, will be highlighted and the devices currently in use will be described. The statistics show the results obtained on 2,500 hemiplegic patients examined for FES application during the last 10 years. The statistics and results of the Slovenian population indicate 0.15-0.20% new cases annually or 1,500 new cases per million inhabitants. Up to 63% of annual cases are candidates for an FES based therapeutic locomotion rehabilitation programme. Experience indicates that 60% of hemiplegic patients received single-channel stimulation to correct equinovarus or foot drop, 30% obtained dual or even three channel stimulation treatment and only 10% of patients were involved in multichannel FES of four to six or even eight channels of stimulation. The benefits and outcome of rehabilitation will be presented and discussed in regard to current trends in the field of FES for hemiplegic and paretic patients. The partly inactive but very important field of FES application to the upper extremity in hemiplegic and paretic patients will be discussed and the relatively modest achievements presented. Future developments will be presented together with advances foreseen by steadily improving technology. PMID- 8233767 TI - Multi-adjustable post-operative orthosis for congenital muscular torticollis. AB - A multi-adjustable torticollis orthosis is described for the post-operative bracing of patients after surgical correction of congenital muscular torticollis. The orthosis can be put on in the early post-operative period and the head and neck position can be maintained in the corrected, and later over-corrected position by the built-in multi-adjustable joint-mechanism. The details of the manufacturing are described. Twenty-five patients (13 girls and 12 boys) from age 1 to 22 with congenital muscular torticollis were fitted with the orthosis post operatively for an average duration of 10 weeks. Satisfactory compliance with the orthosis was found in 23 cases. Complications were minimal (3 cases) and were related to scalp irritation which improved after minor adjustments of the halo. PMID- 8233768 TI - Automatic suspension device for gait training. AB - The automatic suspension device (REHABOT) suspends the patient's body in a standing position allowing the patient to walk around the circular handrail without forward propulsion. Reduction of body weight is accurately maintained automatically while safely supporting the patient. The device was used for 23 patients with orthopaedic disorders or central nervous system disorders who were chosen because of their initial difficulties with gait training in parallel bars. Its advantages are that (1) it may be used for patients with open wounds or cardiac problems, or patients using prostheses or orthoses, (2) preparation and walking practice are simpler both for patients and staff than the therapeutic pool and walking trolley, (3) running costs are lower than the therapeutic pool. Its drawbacks are that the initial cost is relatively high, only one patient can be trained at a time, and the effect of warm water is missing. The automatic suspension device will become one of the new and fundamental pieces of equipment for gait training, especially for hospitals where there are many elderly patients and also severely and multiple disabled persons. PMID- 8233769 TI - Lightweight prostheses for bilateral below-elbow amputees. AB - In view of the anticipated activity of the patient and working environment, lightweight prostheses were designed for an adult female, bilateral below-elbow (BE) amputee at NIRTAR to provide the greatest degree of function. The prostheses were fabricated using lightweight materials and new techniques. Depending on the stump length there were two different types of lightweight prostheses designed and successfully used, (1) an endoskeletal BE prosthesis and (2) an exoskeletal BE prosthesis. After periodic follow-up and evaluation the function of the prostheses was found to be most satisfactory. By reducing the weight considerably compared to other available alternatives, it is more likely that the amputee will make use of the prostheses to efficiently perform various activities. The new prosthesis designs may counteract the high rejection rate of old conventional ones and the principle may be applied to the fabrication of all BE prostheses. PMID- 8233770 TI - Continuous passive motion in hand rehabilitation. AB - This paper reviews the literature comparing the results obtained in applying regimes involving motion with those involving rest following injury or surgery. The deleterious effects of immobilisation are compared to those obtained under conditions of passive motion and intermittent passive motion. It is concluded that continuous passive motion (CPM) represents an improvement on intermittent motion. Models of CPM machines are described and some results presented. It is proposed that "intelligent" CPM would represent a further improvement in technique and a prototype machine for this purpose and some preliminary results are described. PMID- 8233771 TI - The war-injured with physical disabilities are often forgotten in time. PMID- 8233772 TI - Walking speed of normal subjects and amputees: aspects of validity of gait analysis. AB - This study investigated some aspects of the validity of walking speed recording in 15 normal subjects, 16 trans-femoral amputees and 8 knee disarticulation amputees. The variability and test-retest reliability of walking speed and the influence of simultaneous recording of EMG and goniometry on comfortable and fast walking speeds were studied. The variability between sessions was mainly determined by the variance within each session. The variance of speed within sessions while walking with fast speed, was higher when walking without equipment than when walking with equipment. The variances of speed within sessions of the normal subjects were higher than those for both amputee groups. The test-retest reliability, expressed by the intra-class correlation coefficient, was good: between 0.83 and 0.98. The speed when walking without equipment was significantly higher both in normal subjects and amputees than the speed when walking with equipment. PMID- 8233773 TI - Normative ground reaction force data for able-bodied and trans-tibial amputee children during running. AB - The purpose of this investigation was to develop normative ground reaction force data for able-bodied (AB) and trans-tibial amputee (TTA) children during running. Two hundred AB (mean age 9.4 years, range 7-12) and 21 TTA (mean age 11.1 years, range 5-17) children ran (2.2 m/s +/- 10%) over a force platform. Ground reaction force data were normalized, averaged within groups and plotted to produce force time curves characterizing the different leg types (i.e. able-bodied, non prosthetic and prosthetic). In addition, discrete variables characterizing the leg type differences were determined. One way ANOVA determined significant differences between variables and a TukeyB Post Hoc analysis defined which variables were significantly different (p < 0.05). Results generally indicated differences between the three leg types with the non-prosthetic leg indicating greater forces than the prosthetic and AB legs. The results of this investigation provide normative ground reaction force data for both AB and TTA children during running and can be used for comparison with other groups of children. PMID- 8233774 TI - Comparison of gait using a Multiflex foot versus a Quantum foot in knee disarticulation amputees. AB - The subjective responses and gait patterns of unilateral knee disarticulation amputees wearing prostheses fitted first with the Multiflex foot and then with the Quantum foot were studied. Nine amputees were included in the trial. A questionnaire asked the amputees about their preference for one of the feet. Gait analysis was performed measuring temporal parameters and goniometry of hips, knees and ankles in the sagittal and frontal planes. There was a slight preference for the Quantum foot. Preference seemed not to be related to physical characteristics of the amputees nor to gait parameters. There were no differences in gait as far as the temporal factors were concerned. The main differences in the range of motion of the joints were in the frontal plane: the eversion inversion movement of the ankle and the adduction-abduction movement of the hip. During walking at comfortable speed with the Multiflex foot the ankle and hip range of motion averaged 2.1 and 3.1 degrees respectively, less than during walking with the Quantum foot. PMID- 8233775 TI - The CAT-CAM socket and quadrilateral socket: a comparison of energy cost during ambulation. AB - Twenty unilateral trans-femoral amputees fitted with either the Contoured Adducted Trochanteric-Controlled Alignment Method (CAT-CAM) socket (n = 10) or the quadrilateral (QUAD) socket (n = 10), and a "non-amputee" control group (n = 10) participated in the study. Subjects meeting the following criteria were studied: healthy males between the ages of 18 and 55 years, amputation due to non vascular pathology, an unaffected sound limb, at least six months use of the test prosthesis, and a minimal stump length of 15 cm. Subjects ambulated in two randomized trials separated by 20 minutes of rest at 2 assigned speeds: a pace reflecting normal walking speed (97 m/min = 2.5 mph) or a slower speed (48.5 m/min = 1.25 mph). Heart rate (HR) and Oxygen uptake (VO2) measured during steady state walking were analyzed via two-way ANOVA. Differences among means were further analyzed using Tukey post hoc and simple effects tests. Significant differences were observed between the control group and CAT-CAM subjects with respect to VO2 (p < 0.05) and HR (p < 0.01) at the slower speed. The control group and subjects using the QUAD socket also differed with respect to VO2 (p < 0.01) and HR (p < 0.01) at the slower pace. Faster pace required more energy expenditure (p < 0.01) and produced higher HR (p < 0.01) than slower speeds. At faster pace, a significantly higher energy expenditure in the QUAD than the CAT CAM group was observed (p < 0.01). It is concluded that ambulating at normal pace using the CAT-CAM socket design uses less energy than when using a QUAD socket design. PMID- 8233776 TI - A deletion mutant of the type IC restriction endonuclease EcoR1241 expressing a novel DNA specificity. AB - We have developed a complementation assay which allows us to distinguish between mutations affecting subunit assembly and mutations affecting DNA binding in the DNA recognition subunit (HsdS) of the multimeric restriction endonuclease EcoR1241. A number of random point mutations were constructed to test the validity of this assay. Two of the mutants produced were found to be truncated polypeptides that were still capable of complementation with the EcoR1241 Hsd subunits to give an active restriction enzyme of novel DNA specificity. The N terminal variable domain (responsible for recognition of GAA from the EcoR1241 recognition sequence GAAnnnnnnRTCG) and the spacer region (central conserved region) is intact in both of these mutants. One of these mutant genes (hsdS(delta 50) has been cloned as an active Mtase. Purification of the Mtase proved to be difficult because the complex is weak. However, Mtase activity was obtained from a soluble cell extract, and this allowed us to determine the DNA recognition sequence of the Mtase to be GAAnnnnnnnTTC. This recognition sequence is an inverted repeat of 5'-end of the EcoR1241 recognition sequence. This suggests that the mutant Mtase is assembled from two inverted HsdS(D50) subunits, possibly held together by the HsdM subunits. PMID- 8233777 TI - The synthesis of oligoribonucleotides containing O6-methylguanosine: the role of conserved guanosine residues in hammerhead ribozyme cleavage. AB - The synthesis is described of oligoribonucleotides containing the modified nucleoside O6-methylguanosine. Solid-phase oligoribonucleotide assembly was carried out by use of 2'-silyl-protected nucleoside phosphoramidites, a new O6 methylguanosine-containing synthon and a mild deprotection method. The O6 methylguanosine-modified oligonucleotides were used in the study of the role of conserved residues G5, G8 and G12 in hammerhead ribozyme cleavage. Hammerheads thus substituted at any of these positions showed an approximately 75-fold reduction in kcat whereas Km was unaffected. Hammerheads with modifications at G5 or G8 showed a significant reduction in magnesium binding affinity whereas modification at G12 had no effect. The results show that the three conserved G residues play crucial but different role sin hammerhead cleavage. PMID- 8233778 TI - Studies on the synthesis of nucleotidyl-peptides. II. The preparation of a nucleotidyl-peptide having a 5'-nucleotidyl-(P-O)-serine phosphodiester bond. AB - The preparation of a nucleotidyl-peptide having a thymidine-5'-yl-(P-O)-serine phosphodiester bond, [H-Ala-Ser(pTpT)-Phe-OH](24) is described. After condensation between the phosphorylated peptide component and an oligonucleotide component, all protecting groups could be removed under neutral conditions without beta-elimination of the pTpT from the serine moiety. PMID- 8233779 TI - Characterization of gene clusters encoding the fucoxanthin chlorophyll proteins of the diatom Phaeodactylum tricornutum. AB - We are studying the multigene family encoding the fucoxanthin-chlorophyll binding proteins (fcp genes) that constitute the major component of the photosystem II associated light harvesting complex in diatoms and brown algae. The characteristics of clusters of fcp genes on the genome of the diatom Phaeodactylum tricornutum are described. Sequence analysis of two genomic clones, PT5 and PT4, has demonstrated the presence of four fcp genes (fcpA, fcpB, fcpC, fcpD) on the former and two fcp genes (fcpE, fcpF) on the latter. The proteins encoded by the six characterized fcp genes range in similarity from 86% to 99%. The genes within each cluster are separated by short intergenic sequences (between 0.5 to 1.1 kb). None of these genes contain introns and all appear to be transcribed with short 5' transcribed, untranslated leader sequences; the transcription initiation sites were mapped 26 to 48 bases upstream of the ATG translation start site. Small conserved motifs are found among all of the genes just upstream of both the translation and the transcription start sites. The codon bias is similar in all of the fcp genes, with a predominance of pyrimidines in the third positions of codons of the four codon families. The two fcp genes that are most similar are fcpC and fcpD, and might represent a recent gene duplication. Southern analyses using fcp cDNAs as hybridization probes suggest that there may be additional sequences on the P. tricornutum genome that resemble the characterized fcp sequences. PMID- 8233780 TI - Contributions of discrete tRNA(Ser) domains to aminoacylation by E.coli seryl tRNA synthetase: a kinetic analysis using model RNA substrates. AB - The aminoacylation kinetics of T7 transcripts representing defined regions of Escherichia coli serine tRNAs were determined using purified E.coli seryl-tRNA synthetase (SerRS) and the kinetic values were used to estimate the relative contribution of various tRNA(Ser) domains to recognition by SerRS. The analysis revealed that the extra stem/loop structure, characteristic of type II tRNAs such as tRNA(Ser), is the domain which makes the largest contribution to kcat/Km of aminoacylation. Moreover, Km of aminoacylation was increased by a factor of about 1000 when the extra stem/loop was changed to the consensus sequence of type I tRNA extra loops indicating that the stem structure contributes significantly to the binding of tRNA(Ser) to SerRS. A model RNA, which represents only the tRNA(Ser) coaxial acceptor-T psi C stem/loop domain, was also specifically aminoacylated by SerRS having a kcat/Km about 1000-fold greater than background levels. A significant portion of the contribution of this domain to aminoacylation is attributable to the acceptor stem sequence making the acceptor stem the second most important domain for recognition by SerRS. Finally, kcat/Km was essentially unchanged when the entire anticodon stem/loop of tRNA(Ser) was deleted indicating that neither the anticodon nucleotides nor the surrounding stem/loop structure are important for recognition by SerRS. PMID- 8233781 TI - The 5' flanking sequence negatively modulates the in vivo expression and in vitro transcription of a human tRNA gene. AB - The consequences of altering the 5' flanking region of a human amber suppressor tRNA(ser) gene on phenotypic expression in vivo and transcription in vitro was examined by constructing a series of upstream deletion and substitution mutants. The resulting tDNA variants were examined for functional tRNA expression in vivo, by measuring suppression of a nonsense mutation in the Escherichia coli chloramphenicol acetyltransferase (cat) gene in co-transfection assays, and for transcriptional activity in vitro using HeLa cell nuclear extracts. Mutant genes in which the 18 nucleotides 5' proximal to the coding region were deleted and replaced with heterologous sequences were 2 to 5 fold more active in vivo in comparison to the wild type gene. There was a strong, but not exclusive, correlation between the levels of nonsense suppression observed in vivo and transcriptional activity in vitro. In certain cases, introduction of an oligonucleotide encompassing this 18 nucleotide element upstream of more active tRNA genes reduced both the levels of suppression and template activity. These results indicate that the immediate 5' contiguous sequence of this tRNA gene negatively modulates expression both in vivo and in vitro. PMID- 8233782 TI - Efficient method for constructing comprehensive murine Fab antibody libraries displayed on phage. AB - We have developed efficient methodologies for construction and expression of comprehensive phage display libraries of murine Fab antibody fragments in E. coli cells. Our methods optimize several critical steps of the polymerase chain reaction (PCR) amplification of transcripts of the re-arranged immunoglobulin genes and of their subsequent assembly and expression: Firstly, we have designed exhaustive sets of PCR primers of low degeneracy for the amplification of transcripts of the Fab region of the heavy and light-chain genes. These primers proved effective in amplification of Fab gene fragments from a large panel of hybridoma cell lines of different specificity and family sub-type. Secondly, we have developed a 'jumping PCR' technique that effectively assembled and recombined the amplified heavy and light-chain gene fragments into a bi-cistronic operon. Thirdly, we have constructed expression vectors for insertion of the combinatorial Fab gene-cassette in fusion with a truncated version of the phage surface protein, gIIIp. The heavy chain and the light chain-gIII fusion are transcribed as a polycistronic mRNA from the lacZ promoter and efficient transcriptional control is provided by wildtype lacI present on the vector. The utility of the system was demonstrated by isolating several antigen-binding clones from hybridomas and libraries made from immunized mice. PMID- 8233783 TI - Synthesis and hybridization properties of oligonucleotides containing 2'-O modified ribonucleotides. AB - A versatile, general way is described for the introduction of different functional groups into oligonucleotides by means of a simple linker at the 2' position of the sugar. Nucleotide building blocks carrying lipophilic, intercalating or tertiary amino groups can be placed deliberately at any desired position of oligonucleotides by standard automated oligonucleotide synthesis. Thermal denaturation studies with these oligonucleotides reveal the following general trends: i) Modification with lipophilic n-octyl groups has little if any effect on duplex stability; a destabilizing (lipophilic) substituent is better tolerated at or near the ends than in the middle of the oligo. ii) An intercalating substituent (2-aminoanthraquinone) substantially increases duplex stability. iii) N,N-Dimethyl amino residues also increase duplex stability though to a smaller extent than intercalating residues. iv) Modifications at the 5'-end have a more pronounced influence on the TM than the corresponding 3' modifications. v) Oligonucleotides modified in such a way show little or no loss in sequence specificity. PMID- 8233784 TI - Transactivation of the human papilloma virus 16 octamer motif by the octamer binding protein Oct-2 requires both the N and C terminal activation domains. AB - The upstream regulatory region (URR) of the human papillomaviruses HPV16 and 18 contains a sequence with a seven out of eight base match to the consensus binding site for octamer binding transcription factors. This motif acts as a target for repression by the Oct-1 transcription factor and therefore inhibits promoter activity in non-cervical cells expressing only Oct-1. In contrast the HPV octamer motif activates promoter activity in cervical cells. Here we show that cervical cells express the activating form of the Oct-2 transcription factor, Oct 2.1 and that this factor can transactivate promoter activity via the HPV16 octamer. This effect is dependent upon both the N and C-terminal activation domains of Oct-2. The expression of specific octamer binding proteins such as Oct-2 in cervical cells thus allows the HPV16 motif to produce opposite effects on gene expression in cervical and non-cervical cells suggesting that it may play a role in the cervical specificity of URR driven gene expression. PMID- 8233785 TI - Relative specificities in binding of Watson-Crick base pairs by third strand residues in a DNA pyrimidine triplex motif. AB - The specificity of binding of Watson-Crick base pairs by third strand nucleic acid residues via triple helix formation was investigated in a DNA pyrimidine triplex motif by thermal melting experiments. The host duplex was of the type A10 X-A10: T10-Y-T10, and the third strand T10-Z-T10, giving rise to 16 possible triplexes with Z:XY inserts, 4 duplexes with the Watson-Crick base pairs (XY) and 12 duplexes with mismatch pairs (XZ), all of whose stabilities were compared. Two Z:XY combinations confirm the primary binding of AT and GC target pairs in homopurine.homopyrimidine sequences by T and C residues, respectively. All other Z:XY combinations in the T:AT environment result in triplex destabilization. While some related observations have been reported, the present experiments differ importantly in that they were performed in a T:AT nearest neighbor environment and at physiological ionic strength and pH, all of which were previously untested. The conclusions now drawn also differ substantially from those in previous studies. Thus, by evaluating the depression in Tm due to base triplet mismatches strictly in terms of third strand residue affinity and specificity for the target base pair, it is shown that none of the triplet combinations that destabilize qualify for inclusion in the third strand binding code for the pyrimidine triplex motif. Hence, none of the mismatch triplets afford a general way of circumventing the requirement for homopurine.homopyrimidine targets when third strands are predominated by pyrimidines, as others have suggested. At the same time, the applicability of third strand binding is emphasized by the finding that triplexes are equally or much more sensitive to base triplet mismatches than are Watson-Crick duplexes to base pair mismatches. PMID- 8233786 TI - HAPPY mapping of a YAC reveals alternative haplotypes in the human immunoglobulin VH locus. AB - We have identified and sequenced 14 human immunoglobulin VH segments cloned in a yeast artificial chromosome, and have used a rapid PCR-based technique (HAPPY mapping, 12) to derive the order and approximate distances between them. The sequences mapped comprise thirteen germline VH segments and one rearranged VH3 gene. Comparison of our map with other data suggests the existence of at least two distinct haplotypes, differing in the presence or absence of the consecutive genes DP-78, DP-46 and DP-64, and in the duplication of segments DP-49 and DP-65. Screening of ten individuals confirms the existence of both haplotypes, and indicates that both are common amongst the population. PMID- 8233787 TI - An adaptive, object oriented strategy for base calling in DNA sequence analysis. AB - An algorithm has been developed for the determination of nucleotide sequence from data produced in fluorescence-based automated DNA sequencing instruments employing the four-color strategy. This algorithm takes advantage of object oriented programming techniques for modularity and extensibility. The algorithm is adaptive in that data sets from a wide variety of instruments and sequencing conditions can be used with good results. Confidence values are provided on the base calls as an estimate of accuracy. The algorithm iteratively employs confidence determinations from several different modules, each of which examines a different feature of the data for accurate peak identification. Modules within this system can be added or removed for increased performance or for application to a different task. In comparisons with commercial software, the algorithm performed well. PMID- 8233788 TI - Exploiting structural differences among heteroduplex molecules to simplify genotyping the DQA1 and DQB1 alleles in human lymphocyte typing. AB - A novel approach to DNA probe hybridization and heteroduplex analysis, termed directed heteroduplex analysis (DHDA) is presented here to illustrate its utility in simplification of human lymphocyte antigen (HLA)-typing. By strategic labeling of single-stranded probe sequences, DHDA allows the identification of specific heteroduplex structures that contribute to the differentiation of DQA1 and DQB1 alleles. Because of the high degree of polymorphism among major histocompatibility complex class II second exon sequences, this analysis of 50 different heteroduplex molecules provides evidence of the importance of unpaired bases and mismatched base pairs and their effect on heteroduplex electrophoretic mobility differences. This strategy is further used to genotype accurately a family for DQA1 which was previously analyzed by sequence specific oligonucleotide (SSO) probe hybridization. To differentiate by SSO-typing among the DQA1 and DQB1 alleles analyzed in this study requires the use of 23 different probes. Equivalent results are obtained by DHDA using only three probes. Therefore, this study suggests that accurate HLA-typing can be simplified by DHDA. Additionally, DHDA may be useful for differentiation of DNA sequence polymorphisms in other genetic systems. PMID- 8233789 TI - Structures of bulged three-way DNA junctions. AB - We have studied a series of three-way DNA junctions containing unpaired bases on one strand at the branch-point of the junctions. The global conformation of the arms of the junctions has been analysed by means of polyacrylamide gel electrophoresis, as a function of conditions. We find that in the absence of added metal ions, all the results for all the junctions can be accounted for by extended structures, with the largest angle being that between the arms defined by the strand containing the extra bases. Upon addition of magnesium (II) or hexamine cobalt (III) ions, the electrophoretic patterns change markedly, indicative of ion-dependent folding transitions for some of the junctions. For the junction lacking the unpaired bases, the three inter-arm angles appear to be quite similar, suggesting an extended structure. However, the addition of unpaired bases permits the three-way junction to adopt a significantly different structure, in which one angle becomes smaller than the other two. These species also exhibit marked protection against osmium addition to thymine bases at the point of strand exchange. These results are consistent with a model in which two of the helical arms undergo coaxial stacking in the presence of magnesium ions, with the third arm defining an angle that depends upon the number of unpaired bases. PMID- 8233790 TI - Specific binding of MobA, a plasmid-encoded protein involved in the initiation and termination of conjugal DNA transfer, to single-stranded oriT DNA. AB - MobA protein, encoded by the broad host-range plasmid R1162, is required for conjugal mobilization of this plasmid. The protein is an essential part of the relaxosome, and is also necessary for the termination of strand transfer. In vitro, MobA is a nuclease specific for one of the two DNA strands of the origin of transfer (oriT). The protein can cleave this strand at the same site that is nicked in the relaxosome, and can also ligate the DNA. We show here that purified MobA protein forms a complex that is specific for this single oriT strand. The complex is unusually stable, with a half-life of approximately 95 min, is not disrupted by hybridization with the complementary strand, and reforms rapidly after boiling. Both the inverted repeat within oriT, and the eight bases between this repeat and the site cleaved by MobA, are required for binding by the protein. Mutations reducing base complementarity between the arms of the inverted repeat also decrease binding. This effect is partially suppressed by second-site mutations restoring complementarity. These results parallel the effects of these mutations on termination. Footprinting experiments with P1 nuclease indicate that the DNA between the inverted repeat and the nick site is protected by MobA, but that pairing between the arms of the repeat, which occurs in the absence of protein, is partially disrupted. Our results suggest that termination of strand transfer during conjugation involves tight binding of the MobA protein to the inverted repeat and adjacent oriT DNA. This complex positions the protein for ligation of the ends of the transferred strand, to reform a circular plasmid molecule. PMID- 8233791 TI - A pre-export U1 snRNP in Xenopus laevis oocyte nuclei. AB - We demonstrate that precursors of U1 snRNA are associated with nuclear proteins prior to export to the cytoplasm. The approximately 15S complexes containing pre U1 RNA, which we call pre-export U1 snRNPs, were identified in extracts of Xenopus laevis oocyte nuclei that were synthesizing U1 RNAs from injected U1 genes. The U1 snRNP-specific A protein was associated with nuclear pre-U1 RNA since both this protein and the RNA were co-precipitated by antibodies directed against either the m7G-cap of the precursor RNA or the U1-A protein. The interaction of the U1-A protein with pre-U1 RNA required sequences in the loop II region although this region of U1 RNA was not necessary for the association of U1 A protein with mature U1 snRNPs. The U1 A protein helps protect pre-U1 RNA against degradation in the nucleus. PMID- 8233792 TI - Base composition analysis of oligonucleotides containing apurinic sites. AB - Upon base composition analysis, oligonucleotides which are labeled at the 3' terminus with fluorescein or biotin generate an additional, late eluting peak in the HPLC chromatogram. Investigation of this effect revealed the haptens acted as apurinic sites, and phosphodiesterase cleavage of the phosphate bond between the upstream nucleotide and apurinic site is inhibited. Extension of this work with a base-stable apurinic site inserted into all possible junctures of 5'-TGAC-3' tetramers showed this to be a general effect. As a consequence of this work, acid catalyzed depurination resulting in apurinic sites can be monitored in oligonucleotide synthesis. PMID- 8233793 TI - A novel method for the determination of post-transcriptional modification in RNA by mass spectrometry. AB - A method is described for the detection, chemical characterization and sequence placement of post-transcriptionally modified nucleotides in RNA. Molecular masses of oligonucleotides produced by RNase T1 hydrolysis can be measured by electrospray mass spectrometry with errors of less than 1 Da, which provides exact base composition, and recognition of modifications resulting from incremental increases in mass. Used in conjunction with combined liquid chromatography-mass spectrometry and gene sequence data, modified residues can be completely characterized at the nucleoside level, and assigned to sequence sites within oligonucleotides defined by selective RNase cleavage. The procedures are demonstrated using E.coli 5S rRNA, in which all RNase T1 fragments predicted from the rDNA sequence are identified solely on the basis of their molecular masses, and using E.coli 16S rRNA for analysis of post-transcriptional modification, including placement of 3-methyluridine at position 1498. The principles described are generally applicable to other covalent structural modifications of RNA which produce a change in mass, such as those resulting from editing, photochemical cross-linking, or xenobiotic modification. PMID- 8233794 TI - Cloning the RAD51 homologue of Schizosaccharomyces pombe. AB - The RAD51 gene of Saccharomyces cerevisiae encodes a RecA like protein, which is involved in the recombinational repair of double strand breaks. We have isolated the RAD51 homologue, rhp51+, of the distantly related yeast strain Schizosaccharomyces pombe by heterologous hybridization. DNA sequence analysis of the rhp51+ gene revealed an open reading frame of 365 amino acids. Comparison of the amino acid sequences of RAD51 and rhp51+ showed a high level of conservation: 69% identical amino acids. There are two Mlul sites in the upstream region which may be associated with cell cycle regulation of the rhp51+ gene. The rhp51+ null allele, constructed by disruption of the coding region, is extremely sensitive to X-rays, indicating that the rhp51+ gene, like RAD51, is also involved in the repair of X-ray damage. The structural and functional homology between rhp51+ and RAD51 suggests evolutionary conservation of certain steps in the recombinational repair pathway. PMID- 8233795 TI - DNA binding alters the protease susceptibility of the p50 subunit of NF-kappa B. AB - The subdomain structure of the p50 subunit of NF-kappa B (amino acids 35-381) was investigated by partial proteolysis of the native protein. Trypsin cleaves p50 at a limited number of sites with an initial cleavage at low trypsin concentration occurring after R362 and a second cleavage taking place at higher trypsin concentration after K77. The cleavage after R362 does not alter the DNA binding characteristics of p50 but removes the nuclear localisation signal indicating that this region occupies a highly exposed position on the surface of the protein. The second cleavage after K77 generates a protein that although dimeric is incapable of binding DNA, thus emphasising the importance of residues 35-77 in DNA recognition. However p50 dimers containing one molecule cleaved after K77 and one molecule with this region intact are capable of binding DNA. When very high concentrations of trypsin are employed p50 is completely degraded. However if p50 is bound tightly to DNA containing its specific recognition site prior to trypsin addition the cleavage after K77 is almost completely blocked and the protein becomes highly resistant to proteolysis. These data suggest that bound DNA may mask critical trypsin cleavage sites or that DNA binding is accompanied by a conformational change in protein structure that renders the protein resistant to proteolysis. PMID- 8233796 TI - Reduced synonymous substitution rate at the start of enterobacterial genes. AB - Synonymous codon usage is less biased at the start of Escherichia coli genes than elsewhere. The rate of synonymous substitution between E.coli and Salmonella typhimurium is substantially reduced near the start of the gene, which suggests the presence of an additional selection pressure which competes with the selection for codons which are most rapidly translated. Possible competing sources of selection are the presence of secondary ribosome binding sites downstream from the start codon, the avoidance of mRNA secondary structure near the start of the gene and the use of sub-optimal codons to regulate gene expression. We provide evidence against the last of these possibilities. We also show that there is a decrease in the frequency of A, and an increase in the frequency of G along the E.coli genes at all three codon positions. We argue that these results are most consistent with selection to avoid mRNA secondary structure. PMID- 8233797 TI - Dam methyltransferase from Escherichia coli: sequence of a peptide segment involved in S-adenosyl-methionine binding. AB - DNA adenine methyltransferase (Dam methylase) has been crosslinked with its cofactor S-adenosyl methionine (AdoMet) by UV irradiation. About 3% of the enzyme was radioactively labelled after the crosslinking reaction performed either with (methyl-3H)-AdoMet or with (carboxy-14C)-AdoMet. Radiolabelled peptides were purified after trypsinolysis by high performance liquid chromatography in two steps. They could not be sequenced due to radiolysis. Therefore we performed the same experiment using non-radioactive AdoMet and were able to identify the peptide modified by the crosslinking reaction by comparison of the separation profiles obtained from two analytical control experiments performed with 3H AdoMet and Dam methylase without crosslink, respectively. This approach was possible due to the high reproducibility of the chromatography profiles. In these three experiments only one radioactively labelled peptide was present in the tryptic digestions of the crosslinked enzyme. Its sequence was found to be XA GGK, corresponding to amino acids 10-14 of Dam methylase. The non-identified amino acid in the first sequence cycle should be a tryptophan, which is presumably modified by the crosslinking reaction. The importance of this region near the N-terminus for the structure and function of the enzyme was also demonstrated by proteolysis and site-directed mutagenesis experiments. PMID- 8233798 TI - Retroviral-type zinc fingers and glycine-rich repeats in a protein encoded by cnjB, a Tetrahymena gene active during meiosis. AB - We have determined the nucleotide sequence of the cnjB gene from the ciliate Tetrahymena thermophila. This gene is transcriptionally active only during early conjugation, peaking in meiotic prophase. It contains 13 introns, four transcription start points and codes for a putative polypeptide (CnjB) of 1748 amino acids with a calculated molecular weight of 200 kilodaltons and a pl of 7.9. The coding region of cnjB has a low GC content (32% GC) and unusual codon usage. The C-terminal one-third of CnjB consists of three repetitive domains. Introns were absent in this region of cnjB. One of the repetitive domains consists of seven CCHC or retroviral-type zinc fingers, a motif found in one or two copies in retroviral nucleocapsid proteins. This motif has also been found recently in seven copies in the human nucleic-acid binding protein CNBP, in an apparent CNBP homologue in Schizosaccharomyces pombe and in one copy in a Xenopus gene active in early embryos. The other two domains are on either side of the zinc finger domain and contain a repeated glycine-rich motif seen in the heterogeneous nuclear ribonuclear proteins A1 and A2/B1 as well as other proteins. Both CCHC zinc fingers and glycine-rich repeats have been found in proteins with single-stranded nucleic acid-binding activity as well as strand annealing activity. CnjB is, to our knowledge, the first protein found to contain both types of motifs. PMID- 8233799 TI - A more efficient and specific strategy in the ablation of mRNA in Xenopus laevis using mixtures of antisense oligos. AB - Previously, antisense oligodeoxyribonucleotides (oligos) have been used to ablate specific mRNAs from the maternal RNA pool of Xenopus laevis oocytes. However, this strategy is limited by the dose of oligo which can be used and the fact that 100% cleavage of the target RNA is rare. Further, non-specific cleavage of other RNAs can also occur. We demonstrate that the use of several oligos against the histone H4 RNA results in a marked improvement in the efficiency of target degradation, due to synergistic action between oligos and the existence of RNA in at least two different secondary structures. We show, by using a set of overlapping oligos complementary to the entire H4 RNA, that the amount of oligo required for efficient target ablation is greatly lowered and non-specific effects are reduced. PMID- 8233800 TI - Encapsidation of heterologous RNAs by bacteriophage MS2 coat protein. AB - The RNA bacteriophages of E. coli specifically encapsidate a single copy of the viral genome in a protein shell composed mainly of 180 molecules of coat protein. Coat protein is also a translational repressor and shuts off viral replicase synthesis by interaction with a RNA stem-loop containing the replicase initiation codon. We wondered whether the translational operator also serves as the viral pac site, the signal which mediates the exclusive encapsidation of viral RNA by its interaction with coat protein. To test this idea we measured the ability of lacZ RNA fused to the translational operator to be incorporated into virus-like particles formed from coat protein expressed from a plasmid. The results indicate that the operator-lacZ RNA is indeed encapsidated and that nucleotide substitutions in the translational operator which reduce the tightness of the coat protein-operator interaction also reduce or abolish encapsidation of the hybrid RNA. When coat protein is expressed in excess compared to the operator lacZ RNA, host RNAs are packaged as well. However, elevation of the level of operator-lacZ RNA relative to coat protein results in its selective encapsidation at the expense of cellular RNAs. Our results are consistent with the proposition that this single protein-RNA interaction accounts both for translational repression and viral genome encapsidation. PMID- 8233801 TI - The interaction between the iron-responsive element binding protein and its cognate RNA is highly dependent upon both RNA sequence and structure. AB - To assess the influence of RNA sequence/structure on the interaction RNAs with the iron-responsive element binding protein (IRE-BP), twenty eight altered RNAs were tested as competitors for an RNA corresponding to the ferritin H chain IRE. All changes in the loop of the predicted IRE hairpin and in the unpaired cytosine residue characteristically found in IRE stems significantly decreased the apparent affinity of the RNA for the IRE-BP. Similarly, alteration in the spacing and/or orientation of the loop and the unpaired cytosine of the stem by either increasing or decreasing the number of base pairs separating them significantly reduced efficacy as a competitor. It is inferred that the IRE-BP forms multiple contacts with its cognate RNA, and that these contacts, acting in concert, provide the basis for the high affinity of this interaction. PMID- 8233802 TI - SnoI, a novel alternatively spliced isoform of the ski protooncogene homolog, sno. AB - We have cloned and sequenced a novel human isoform of sno, snoI for insertion. SnoI contains 1330 nucleotides inserted in place of 7 nucleotides of the snoN mRNA. Sno is a member of the ski protooncogene family, which has been implicated in muscle development. The two previously known sno alternatively spliced isoforms are snoN (684 amino acids), and snoA (415 amino acids); snoI encodes a truncated isoform of 399 amino acids (44,298 MW). Southern blot experiments show that snoI contains a third alternative exon from the sno gene; a single sno gene can express all three isoforms of sno by alternative splicing. All three isoforms contain the region that is most similar to the ski proto-oncogene. The relationship between snoI and snoN is analogous to that between delta fosB and fosB, where a truncated form of the fosB transcription factor is produced by alternative splicing. We find conservation of human snoI-specific sequences in several mammalian species, in monkey, dog, cow, rabbit and pig, but not in rodents, whereas the common portion of the sno gene is conserved in all vertebrate species tested. SnoN, snoA, and ski mRNAs accumulate in many human tissues including skeletal muscle; the snoI alternative mRNA accumulates more specifically in skeletal muscle. SnoI is also expressed in rhabdomyosarcoma tumor, a tumor that contains differentiated skeletal muscle. The tissue-specific alternative splicing of human snoI, an mRNA in the ski/sno gene family, and the presence of sno mRNAs in muscle are consistent with a proposed role for the sno oncogene in muscle gene regulation. PMID- 8233803 TI - Isolation and characterization of cDNAs encoding the sea urchin (Strongylocentrotus purpuratus) homologue of the CCAAT binding protein NF-Y A subunit. PMID- 8233804 TI - Complete nucleotide sequence of Candida albicans 5.8S rRNA coding gene and flanking internal transcribed spacers. PMID- 8233805 TI - A method for creating chromosome-specific plasmid libraries enriched in clones containing [CA]n microsatellite repeat sequences directly from flow-sorted chromosomes. PMID- 8233806 TI - Temperature-gradient gel electrophoresis for the detection and characterization of curved DNA-fragments. PMID- 8233807 TI - A rapid assay for quantitative detection of specific RNAs. PMID- 8233808 TI - Different thermostable DNA polymerases may amplify different RAPD products. PMID- 8233809 TI - Solution hybridization with Hybond-mAP paper. PMID- 8233810 TI - Branched poly-labelled oligonucleotides: enhanced specificity of fork-shaped biotinylated oligoribonucleotides for antisense affinity selection. PMID- 8233811 TI - Chromatin analysis in yeast using NP-40 permeabilised sphaeroplasts. PMID- 8233812 TI - Methylation analysis by means of PCR SSCP: application to clonality studies. PMID- 8233813 TI - Direct PCR from solid tissues without DNA extraction. PMID- 8233814 TI - Conserved sequence motif DPPY in region IV of the phage T4 Dam DNA-[N6-adenine] methyltransferase is important for S-adenosyl-L-methionine binding. AB - Comparison of the deduced amino acid sequences of DNA-[N6-adenine] methyltransferases has revealed several conserved regions. All of these enzymes contain a DPPY [or closely related] motif. By site-directed mutagenesis of a cloned T4 dam gene, we have altered the first proline residue in this motif [located in conserved region IV of the T4 Dam-MTase] to alanine or threonine. The mutant enzymic forms, P172A and P172T, were overproduced and purified. Kinetic studies showed that compared to the wild-type [wt] the two mutant enzymic forms had: (i) an increased [5 and 20-fold, respectively] Km for substrate, S-adenosyl methionine [AdoMet]; (ii) a slightly reduced [2 and 4-fold lower] kcat; (iii) a strongly reduced kcat/KmAdoMet [10 and 100-fold]; and (iv) almost the same Km for substrate DNA. Equilibrium dialysis studies showed that the mutant enzymes had a reduced [4 and 9-fold lower] Ka for AdoMet. Taken together these data indicate that the P172A and P172T alterations resulted primarily in a reduced affinity for AdoMet. This suggests that the DPPY-motif is important for AdoMet-binding, and that region IV contains or is part of an AdoMet-binding site. PMID- 8233815 TI - A study of oligonucleotide reassociation using large arrays of oligonucleotides synthesised on a glass support. AB - An extensive analysis of oligonucleotide interactions was carried out by hybridising a synthetic pool of 256 10mers, A(C,T)8A, representing all oligopyrimidine octamer sequences to an array of four copies of all 256 different octapurine sequences. The resulting 256 duplexes were quantified by phosphorimaging and analysed to determine the dependence of duplex formation on base composition, sequence, and salt concentration. The results show that the base composition dependence of duplex formation can be reduced by high concentrations of tetramethylammonium chloride. This chaotropic solvent also increases duplex yield by up to fifty-fold. PMID- 8233816 TI - Conformational analysis of the 5' leader and the gag initiation site of Mo-MuLV RNA and allosteric transitions induced by dimerization. AB - Dimerization of genomic RNA is a key step in the retroviral life cycle and has been postulated to be involved in the regulation of translation, encapsidation and reverse transcription. Here, we have derived a secondary structure model of nucleotides upstream from psi and of the gag initiation region of Mo-MuLV RNA in monomeric and dimeric forms, using chemical probing, sequence comparison and computer prediction. The 5' domain is extensively base-paired and interactions take place between U5 and 5' leader sequences. The U5-PBS subdomain can fold in two mutually exclusive conformations: a very stable and extended helical structure (E form) in which 17 of the 18 nucleotides of the PBS are paired, or an irregular three-branch structure (B form) in which 10 nucleotides of the PBS are paired. The dimeric RNA adopts the B conformation. The monomeric RNA can switch from the E to the B conformation by a thermal treatment. If the E to B transition is associated to dimerization, it may facilitate annealing of the primer tRNAPro to the PBS by lowering the free energy required for melting the PBS. Furthermore, dimerization induces allosteric rearrangements around the SD site and the gag initiation region. PMID- 8233817 TI - Artificial self-cleaving molecules consisting of a tRNA precursor and the catalytic RNA of RNase P. AB - We synthesized two types of chimeric RNAs between the catalytic RNA subunit of RNase P from Escherichia coli (M1 RNA) and a tRNA precursor (pre-tRNA); one had pre-tRNA at the 3' side to the M1 RNA (M1 RNA-pre-tRNA). The second had pre-tRNA at the 5' side of the M1 RNA (pre-tRNA-M1 RNA). Both molecules were self-cleaving RNAs. The self-cleavage of M1 RNA-pre-tRNA occurred at the normal site (5'-end of mature tRNA sequence) and proceeded under the condition of 10 mM Mg2+ concentration. This reaction at 10 mM Mg2+ was an intramolecular reaction (cis cleavage), while, at 40 mM and 80 mM Mg2+, trans-cleavage partially occurred. The self-cleavage rate was strictly affected by the distance between the M1 RNA and the pre-tRNA in the molecule. The self-cleavage of pre-tRNA-M1 RNA occurred mainly at three sites within the mature tRNA sequence. This cleavage did not occur at 10 mM Mg2+. Use of M1 RNA-pre-tRNA molecule for the in vitro evolution of M1 RNA is discussed. PMID- 8233818 TI - 3' truncated tRNAArg is essential for in vitro specific cleavage of partially synthesized mouse 18S rRNA. AB - In vitro synthesized 5' portions of mouse 18S rRNA are cleaved efficiently at a specific site in partially purified extracts of mouse FM3A cells and several mouse tissues. This activity is composed of both protein and RNA, and can be reconstituted with the protein component in micrococcal nuclease-treated extracts and the RNA component in phenol-treated ones. The RNA component of about 65 nucleotides with the complementing activity was purified from total RNA in the partially purified FM3A cell extracts by polyacrylamide gel electrophoreses. Chemical sequencing of this RNA elucidated that it is tRNAArg lacking nine nucleotides from its 3' terminus. Ribonuclease H treatment directed by deoxyoligonucleotides complementary to tRNAArg completely abolishes the cleavage activity, supporting the above conclusion. PMID- 8233819 TI - Coordinate translational regulation in the syntheses of elongation factor 1 alpha and ribosomal proteins in Xenopus laevis. AB - The regulation of the synthesis of elongation factor 1 alpha (EF-1 alpha) in Xenopus laevis has been analyzed from the point of view of translational control. The 5' end of EF-1 alpha mRNA, examined by primer extension, revealed the presence of a terminal pyrimidine tract that is characteristic of ribosomal protein mRNAs (rp-mRNAs). We have then compared the translation pattern of EF-1 alpha and rp-mRNAs during Xenopus embryogenesis and in Xenopus cultured cells during growth rate changes. In Xenopus embryos EF-1 alpha transcripts, that appear after midblastula transition, are initially mostly localized on mRNP and translationally inactive. Only later in embryogenesis, together with rp-mRNAs, they are gradually recruited on polysomes. Also in Xenopus cells B 3.2, EF-1 alpha mRNA shows a distribution change similar to an rp-mRNA: part of it moves from polysomes to mRNP during serum deprivation and goes back on polysomes after restitution of serum to the culture. Moreover EF-1 alpha mRNA, similarly to rp mRNAs, is always localized on mRNP or fully loaded on polysomes but never on small polysomes. Therefore EF-1 alpha mRNA for structural features and translation behavior can be included in the 'regulatory' group of rp-mRNAs. PMID- 8233820 TI - Cpf1 protein induced bending of yeast centromere DNA element I. AB - The centromere complex is a multicomponent structure essential for faithful chromosome transmission. Here we show that the S. cerevisiae centromere protein Cpf1 bends centromere DNA element I (CDEI) with the bend angle ranging from 66 degrees to 71 degrees. CDEI DNA sequences that carry point mutations which lead to reduced Cpf1 binding affinity and in vivo centromere activity are still able to show bending. The Cpf1 induced bend is directed towards the major groove with the bend centre located in CDEI. An intrinsic bend cannot replace the Cpf1 induced DNA bend for in vivo centromere function. An in vivo phasing experiment suggests that both the distance and the correct spatial arrangement of the CDEI/Cpf1 complex to CDEII and CDEIII are important for optimal centromere function. PMID- 8233821 TI - Nucleosomal structure at hyperacetylated loci probed in nuclei by DNA-histone crosslinking. AB - Chemically induced histone-DNA crosslinking in nuclei is used to monitor structural changes in chromosomal domains containing hyperacetylated histones. Core particles harbouring the crosslinks are immunofractionated with antibodies specific for acetylated histones. Crosslinking is revealed by gel separation of tryptic peptides from core histones that carry 32P-labelled residual nucleotide. The large number of DNA-histone crosslinks retained indicates that acetylated core histone tails are not totally displaced from the DNA. Changes in the patterns of crosslinked peptides imply a restructuring of hyperacetylated histone DNA interactions at several points within the nucleosome. This demonstrates that a distinct conformational state is adopted in acetylated nucleosomes, known to be concentrated at transcriptionally active loci. PMID- 8233822 TI - Effect of excess water on the desilylation of oligoribonucleotides using tetrabutylammonium fluoride. AB - The most commonly available 2' hydroxyl protecting group used in the synthesis of oligoribonucleotides is the tert-butyldimethylsilyl moiety. This protecting group is generally cleaved with 1 M tetrabutylammonium fluoride (TBAF) in tetrahydrofuran (THF). The efficiency of this reaction was tested on ribonucleotidyldeoxythymidine dinucleotides (AT, CT, GT, and UT). We have found that the efficiency of desilylation of uridine and cytidine is greatly dependent on the water content of the TBAF reagent. Conversely, the water content of the TBAF reagent [up to 17% (w/w)] had no detectable effect on the rate of desilylation of adenosine and guanosine. It was concluded that for effective desilylation of pyrimidine nucleosides the water content of the TBAF reagent must be 5% or less, which is readily achieved using molecular sieves. TBAF dried in such a manner was shown to be effective in deprotecting an oligoribonucleotide containing both purine and pyrimidine residues. PMID- 8233823 TI - In vivo analysis of Pim-1 deficiency. AB - The Pim-1 proto-oncogene encodes a highly conserved serine/threonine phosphokinase which is predominantly expressed in hematopoietic organs and gonads in mammals. Overexpression of Pim-1 predisposes to lymphomagenesis in mice. To develop a further understanding of Pim-1 in molecular terms, as well as in terms of its potential role in hematopoietic development, we have generated mice deficient in Pim-1 function. Pim-1-deficient mice are ostensibly normal, healthy and fertile. Detailed comparative analyses of the hematopoietic systems of the mutant mice and their wild-type littermates showed that they are indistinguishable for most of the parameters studied. Our analyses revealed one unexpected phenotype that correlated with the level of Pim-1 expression: Pim-1 deficiency correlated with a erythrocyte microcytosis, whereas overexpression of Pim-1 in E mu-Pim-1-transgenic mice resulted in erythrocyte macrocytosis. In order to confirm that the observed decrease in erythrocyte Mean Cell Volume (MCV) was attributable to the Pim-1 deficiency, we developed mice transgenic for a Pim 1 gene construct with its own promoter and showed that this transgene could restore the low erythrocyte Mean Cell Volume observed in the Pim-1-deficient mice to near wild-type levels. These results might be relevant to the observed involvement of the Pim-1 gene in mouse erythroleukemogenesis. The surprising lack of a readily observed phenotype in the lymphoid compartment of the Pim-1 deficient mice, suggests a heretofore unrecognized degree of in vivo functional redundancy of this highly conserved proto-oncogene. PMID- 8233824 TI - Methylphosphate cap structure increases the stability of 7SK, B2 and U6 small RNAs in Xenopus oocytes. AB - We studied the role of the methylphosphate cap structure in the stability and nucleocytoplasmic transport by microinjecting U6, 7SK and B2 RNAs into the Xenopus oocytes. In every case, the methylphosphate capped RNAs were 3 to 9 times more stable than the uncapped RNAs. When a methylphosphate cap structure was placed on human H1 RNA which is normally not capped, its stability was improved 2 7 fold. These data show that the methylphosphate cap enhances the stability of 7SK, B2, H1 and U6 RNAs. The methylphosphate-capped 7SK RNA was transported into the nucleus from cytoplasm, but remained in the nucleus when injected into the nucleus; in this respect, 7SK RNA exhibited properties previously shown for U6 RNA. Both U6 and 7SK RNAs with ppp on their 5' ends were transported from cytoplasm to the nucleus suggesting that the methylphosphate cap structure is not required for transport of these RNAs across the nuclear membrane. PMID- 8233825 TI - cis-elements involved in alternative splicing in the rat beta-tropomyosin gene: the 3'-splice site of the skeletal muscle exon 7 is the major site of blockage in nonmuscle cells. AB - We have been using the rat beta-tropomyosin (beta-TM) gene as a model system to study the mechanism of alternative splicing. The beta-TM gene spans 10 kb with 11 exons and encodes two distinct isoforms, namely skeletal muscle beta-TM and fibroblast TM-1. Exons 1-5, 8, and 9 are common to all mRNAs expressed from this gene. Exons 6 and 11 are used in fibroblasts, as well as in smooth muscle cells, whereas exons 7 and 10 are used exclusively in skeletal muscle cells. Our previous studies localized the critical elements for regulated alternative splicing to sequences within exon 7 and the adjacent upstream intron. We also demonstrated that these sequences function, in part, to regulate splice-site selection in vivo by interacting with cellular factors that block the use of the skeletal muscle exon in nonmuscle cells (1). Here we have further characterized the critical cis-acting elements involved in alternative splice site selection. Our data demonstrate that exon 7 and its flanking intron sequences are sufficient to regulate the suppression of exon 7 in nonmuscle cells when flanked by heterologous exons derived from adenovirus. We have also shown by both in vivo and in vitro assays that the blockage of exon 7 in nonmuscle cells is primarily at its 3'-splice site. A model is presented for regulated alternative splicing in both skeletal muscle and nonmuscle cells. PMID- 8233826 TI - A correlation between dexamethasone inducibility and basal expression levels of retroviral vector proviruses. AB - Identical transcription units inserted at different positions of mammalian chromosomes may vary widely in transcriptional activity. We have used a set of ten cell clones with random unselected single integrations of retroviral vectors to study such position effects. The vector used carries a neo gene driven by the Akv murine leukemia virus long terminal repeat that has only a weak promoter enhancer activity in the target cell, the lymphoid cell line L691. Under transient expression conditions, the strength of the Akv promoter-enhancer in the L691 cells is increased by dexamethasone. In cell clones with single vector integrations, a correlation is observed between the non-induced expression levels and the degree of dexamethasone induction. The strongest relative induction is found for the integrated vectors with the lowest non-induced expression levels and approaches the inducibility under transient expression. These results indicate that expression levels are composed of distinct contributions from the integrated vector and from the site of integration and are best explained in terms of a model in which the sites of chromosomal integration exert variable positive enhancer effects upon vector transcription. PMID- 8233827 TI - A method for the generation of YAC transgenic mice by pronuclear microinjection. AB - Yeast artificial chromosomes (YACs) represent the latest generation of vectors which have the great advantage of large insert size. The introduction of YACs into mammalian cells and organisms has become an important goal, since it offers the potential to study the control of large and complex transcription units and identify genes by complementation. Microinjection into the nucleus is the most direct and efficient way of delivering YAC DNA into cells, but requires the purification of the YAC from the remaining yeast chromosomes. Here we describe a detailed method for the isolation of pure, intact and highly concentrated YAC DNA. As a model system the murine tyrosinase gene was chosen and four YACs covering this locus were isolated. Introduction by homologous recombination in yeast of sequences permitting YAC amplification greatly facilitated the isolation of YAC DNA at high concentrations. YAC DNA stabilized in a salt and polyamine containing buffer did not compromise the survival of microinjected oocytes and was suitable for the generation of transgenic mice. Applications and benefits of this technique will be discussed. PMID- 8233828 TI - Mapping and positioning DNA-binding proteins along genomic DNA. Structure of D. melanogaster ribosomal 'Alu-repeats' and 1.688 satellite chromatin. AB - Chromatin structure of so-called 'Alu-repeat' in D. melanogaster ribosomal non transcribed spacer that contains sequences homologous to the promoter of ribosomal genes has been studied. Using the 'protein image' hybridization assay based on UV-light-induced DNA-protein crosslinking and 2-D gel retardation electrophoresis, two proteins of the molecular mass of 50 kD (rABP50) and 70 kD (rABP70), associated with 'Alu-repeat' DNA have been found. Exo III mapping of crosslinking sites and DNase I footprinting have provided a detailed map of H1, rABP50 and rABP70 contacts within the 'Alu-repeat' and H1 and a non-histone protein contacts on satellite DNA. These data indicate precise positioning of non histone proteins, histone H1 and nucleosomes within genomic regions studied and account for the presence of unusual 240 bp long nucleosomal particles in 'Alu repeats'. The same approach can be adapted for successive mapping and positioning proteins on genomic DNA. PMID- 8233829 TI - A novel transforming growth factor-beta 1 responsive cytoplasmic trans-acting factor binds selectively to the 3'-untranslated region of mammalian ribonucleotide reductase R2 mRNA: role in message stability. AB - Ribonucleotide reductase is a highly regulated enzyme that provides the four deoxyribonucleotides required for DNA synthesis. Our studies showed that TGF-beta 1 treatment of BALB/c 3T3 mouse fibroblasts markedly elevated ribonucleotide reductase R2 mRNA levels, and also increased the half-life of R2 message by 4 fold from 1.5 h in untreated cells to 6 h in treated cells. We describe a novel 75 Kd sequence-specific cytoplasmic factor (p75) that binds selectively to a 83 nucleotide 3'-untranslated region of R2 mRNA and did not bind to the 5'UTR, the coding region of the R2 message or to the 3'UTRs of other mRNAs (from c-myc, GM CSF and the iron responsive element from the transferrin receptor mRNA), or to the homopolymer poly(A) sequence. p75-RNA binding activity, which requires new protein synthesis, is not present in untreated cells, but is induced following TGF-beta 1 stimulation. The in vivo kinetics of appearance of p75 binding activity paralleled the accumulation of R2 mRNA. Insertion of the 3'-untranslated region into the chloramphenicol acetyltransferase (CAT) message confers TGF-beta 1 induced stability of RNA in stably transfected cells, while the same insert carrying a deletion of the 83-nucleotide fragment had little affect on RNA levels. Furthermore, in vitro decay reactions that contained the 83-nucleotide RNA or deletion of this fragment caused a significant decrease in TGF-beta 1 stabilization of R2 message. A model is presented of R2 message regulation in which TGF-beta 1 mediated stabilization of R2 message involves a specific interaction of a p75-trans-acting factor with a cis-element(s) stability determinant within the 83-nucleotide sequence which is linked to a reduction in the rate of R2 mRNA degradation. PMID- 8233830 TI - Triple helix formation at distant sites: hybrid oligonucleotides containing a polymeric linker. AB - An oligonucleotide hybrid is described which possesses two triple helix forming oligonucleotides which have been connected by a flexible polymeric linker chain. As a prototype, binding of this class of oligonucleotide to duplex DNA has been studied using a segment of the HSV-1 D-glycoprotein promoter, which possesses a pair of 12bp target sites for stable triple helix formation, separated by a duplex spacer region which is one helical turn long. Band shift and footprinting analysis show that such hybrids can bind to both 12bp elements simultaneously, if flexible linkers are included which are longer than 20-25 rotatable bonds. Molecular modeling confirms that a flexible polymeric linker as short as 22 rotatable bonds is enough to link the two distant segments of triple helix, providing that the linker element travels a path which is external to the helix grooves and parallel to the long helix axis. PMID- 8233831 TI - Mapping of replication initiation site in Mycoplasma capricolum genome by two dimensional gel-electrophoretic analysis. AB - The homolog of the dnaA gene, which has been reported to be present in the vicinity of the initiation site of replication in the genome of Mycoplasma capricolum (M.Miyata, L.Wang, and T.Fukumura, J. Bacteriol. 175: 655-660, 1993) was mapped precisely. A 9540-bp region containing the dnaA gene was cloned and the entire region was sequenced with the exception of a previously reported region of 2517 bp (Fujita, M.Q., Yoshikawa, H. and Ogasawara, N. Gene 93: 73-78, 1992). The organization of the 9540-bp region was compared with that of corresponding regions in other bacteria. The arrangement and directions of rnpA, rpmH, dnaA, dnaN were conserved, but no other open reading frames were found that were homologous to those that are commonly found around dnaA genes in other bacteria. The directions of movement of the replication fork around the dnaA gene were analyzed by neutral/alkaline two-dimensional gel electrophoresis. The forks developed in a 1569-bp region that consisted of the dnaA structural gene and its downstream non-coding region, and then they proceeded bidirectionally. PMID- 8233832 TI - In vivo footprinting of the human IL-2 gene reveals a nuclear factor bound to the transcription start site in T cells. AB - The IL-2 gene is a T cell specific gene that is expressed early during the activation-specific T lymphocyte development program. Electrophoretic mobility shift assay (EMSA) and DNase I footprinting assays have defined DNA/protein interactions at the IL-2 promoter cis-elements in vitro. To determine if the trans-activators documented in T cell nuclear extracts actually bind the IL-2 promoter in vivo, ligation mediated PCR (LMPCR) genomic footprinting was performed on the IL-2 promoter in both activated and non-activated T cells and HL60 promyelocytes, which do not express the IL-2 gene. The in vivo footprints indicate that the IL-2 gene transcription start site and TATA sequence are protected in both activated and resting T cells, prior to the appearance of detectable IL-2 steady state message. The distal NF-AT and the NF kappa B sites are each footprinted and the Oct/OAP site contains hypersensitive residues in the unstimulated T lymphocytes. Additional residues are protected in each of these sites after T cell activation. The proximal NF-AT site (NF-IL-2B) and the AP-1 site at -150 are protected in activated Jurkat T lymphocytes, but these two sites are not protected in activated Jurkat lymphocytes stably transfected a gene construct containing multiple NFAT binding sites. PMID- 8233833 TI - Recombinant retroviral systems for the analysis of drug resistant HIV. AB - Two recombinant retroviral systems are described that can be used to analyze antiretroviral drug activity and HIV breakthrough (replication in the presence of the drug). The first system utilizes a recombinant HIV encoding beta galactosidase as a reporter gene (HIV-LacZ). The defective HIV-LacZ virus is produced in COS cells after co-transfection of a plasmid encoding the HIV-LacZ genome with a plasmid encoding HIV proteins necessary for packaging and infectivity. Subsequent infection of CD4+ target cells, followed by assay for LacZ expression, permits the rapid identification of individual virus-infected cells. This system can be used to quantitate the inhibition of early events in the HIV replicative cycle and is suitable for the screening of compounds for anti HIV activity. However, this system cannot be used to analyze HIV drug resistance because of the limited genetic heterogeneity of the virus that is produced in COS cells. To circumvent this problem, a second system has been developed in which heterogenous recombinant HIV is produced by rescue with replication-competent 'helper' HIV. This system required the production of CD4+ cell lines containing defective proviruses encoding either LacZ or guanosine phosphoribosyl transferase (gpt). The defective proviruses are rescued by infection of the cell lines with 'helper' HIV and used to infect target cells in the presence of antiretroviral agents. Subsequent reporter gene assay is used to identify virus-infected cells. This system has been used to detect rare HIV breakthrough infection of cells in the presence of the non-nucleoside reverse transcriptase inhibitor TIBO R82150. Similar analyses with other antiretroviral agents, alone and in combination, may help identify therapeutic strategies that minimize breakthrough replication of HIV. PMID- 8233834 TI - Isolation and characterization of the modification methylase M.SauLPI from Streptomyces aureofaciens B-96. PMID- 8233835 TI - The complete murine cDNA sequence of the transcription factor AP-2. PMID- 8233836 TI - Nucleotide sequence of the small subunit rRNA of Septata intestinalis. PMID- 8233837 TI - A highly conserved sequence motif defining the family of MutT-related proteins from eubacteria, eukaryotes and viruses. PMID- 8233838 TI - Rapid purification of high-activity Taq DNA polymerase. PMID- 8233839 TI - PCR priming from the restriction endonuclease site 3' extension. PMID- 8233840 TI - [Functional scintigraphy in the follow-up of transplants]. AB - Nuclear medicine procedures have special indications in the follow-up of transplanted patients: In case of renal transplantation perfusion, function (glomerular filtration) and urinary flow measured by bolus application of 99mTc DTPA is an important tool to detect and monitor acute tubular necrosis, rejection, urinary leakage, obstruction or vascular complications. This method is used for pancreatic grafts, too. To exclude rejection in case of heart transplantation more than one year after operation the antimyosin antibody scintigraphy is introduced avoiding biopsies. The radionuclide ventriculography is able to monitor heart function by determining the ejection fraction. Cholescintigraphy in liver transplants is sensitive to detect complications of bile flow. In case of an auxiliary liver transplantation a competition between two livers has been described. This method allows a distinction of functional performance of donor and recipient liver. The same problem is solved by ventilation-perfusion scan in lung transplants. Bone scintigraphy is of prognostic value for graft viability. Other radionuclide examinations have been developed to measure perfusion, glucose-, fat-metabolism to detect rejection episodes, abscesses or tumor recurrencies. PMID- 8233841 TI - Gated blood pool SPECT and phase analysis to assess simulated Wolff-Parkinson White syndrome in the baboon. AB - This study assesses the diagnostic potential of a tomographic technique with phase analysis to detect premature electroventricular contraction patterns simulated by pacing in the baboon. The data of gated SPECT were analysed by backprojection of the Fourier coefficients, followed by angulation and integration to thick slices of the entire ventricular mass yielding separate ventricular contraction patterns in three perpendicular views. Electrodes were implanted in each baboon: at the sinoatrial node; posterior, left ventricular; anterior left ventricular; on the left and the right lateral ventricular walls. The atrium was stimulated throughout at a fixed rate. Subsequent ventricular stimuli followed during the QRS complex, such to invoke the appearance of pre excitation QRS morphology. The first points of activation (FPA) from this algorithm were correctly detected for the RV, for the anterior and posterior sites, although the latter two manifested first points in the RV. LV pacing manifested also as a FPA in the RV, but was followed by a true subsequent point in the LV. PMID- 8233842 TI - [Brain SPECT with 123I-labelled iodobenzamide (IBZM): aspects of semiquantitative evaluation]. AB - The striatal dopamine-D2-receptor uptake of 123I-IBZM in 32 patients (18 without and 14 under therapy with typical neuroleptics) was measured semiquantitatively using different ROI techniques. The aim of the study was to evaluate the influence of these techniques on the different ratios (striatum/cortex) found by various examiners. Using the same SPECT system no major differences were found between rectangular and manually drawn ROIs of the same size. All ROI techniques could differentiate between patients with and without therapy on a highly significant level. Therefore, the resulting ratio is mainly dependent on the spatial resolution of the camera system and only to a relatively minor extent on the ROI technique. Ratios obtained by different observers are not comparable quantitatively. PMID- 8233843 TI - Quantitative bone scintigraphy in prostatic carcinoma--long-term response to treatment. AB - Quantitative bone scintigraphy was performed in 24 patients with prostatic carcinoma before orchiectomy and up to one to four years after operation. The gamma camera count rate was recorded over the lower thoracic and all lumbar vertebrae 4 h after injection of 99mTc-MDP. Twelve patients had normal bone scintigrams throughout the study. They showed from two years after operation a slight increase in count rate values compared with the preoperative values, probably due to hormonal changes after orchiectomy and to age-related alterations in skeletal metabolism. Twelve patients had abnormal bone scintigrams. They showed as a response to treatment the flare phenomenon with an increase in count rate over the abnormal vertebrae when measured two weeks after operation followed by a decrease after two months. The lowest count rate values were obtained between six months and one year after operation. Thereafter the count rate seemed to remain on the same level. An increase in count rate was connected to progression of disease and the patients died of prostatic carcinoma within one year thereafter. PMID- 8233844 TI - [Approximation errors in the physical planning of radioiodine therapy of the thyroid]. AB - The approximations in the equations used for the calculation of the radioiodine therapy are reviewed and compared with a reference formula, based on ICRP Publication 30. Applying these equations to two clinical cases, we find pronounced differences compared to the reference method. Investigations of a collective of 314 patients yield in a relatively high fraction delayed maxima in the uptake curves. Simulations prove that serious errors occur when the ascending slope of the uptake curve is neglected. An equation taking care of this effect is presented. Pronounced individual differences are found between uptake curves recorded before and after therapy. The effects on the difference between planned and actually applied dose to the thyroid are shown. The difficulties of the determination of the thyroid mass are discussed and a combined method using scinti- and sonographic data presented. PMID- 8233845 TI - [Highlights and trends--German Society of Nuclear Medicine, Annual Meeting 1993 in Cologne]. AB - The lecture presented at the end of the annual meeting 1993 of the German Nuclear Medicine Society in Cologne contrasts with the usual form of "high-light lecture". The aim was to elucidate developments and to analyse trends in nuclear medicine between the '87 congress in Freiburg and the '93 meeting. This analysis covered the following topics: In-vitro tests, magnetic resonance imaging, therapy with radioisotopes, "newly" developed tracers, methods and their comparison, correlative studies, interventional activities, relationship of the presented papers to daily clinical routine and, finally, scope and size of the study populations. Only few scientific contributions were mentioned to support the trend analysis presented. PMID- 8233846 TI - Fatty tissue visualization 18 hours after 133Xe lung ventilation study. AB - In order to check a new rebreathing unit for ventilation investigations of the lungs with radioactive gases, a 133Xe lung ventilation study was performed in a 56-year old healthy volunteer. The contamination check on the following day triggered a radiation warning caused by the retained xenon activity in the body. The whole-body scan performed 18 h after the inhalation showed clearly a faint delineation of the xenon activity, corresponding to the subcutaneous fatty tissue of the individual studied. PMID- 8233847 TI - [The value of scintigraphic functional assessment in extreme hydronephrosis]. AB - Renographic studies are occasionally suspected of overestimating the excretory function of obstructed kidneys. Presenting the case of a ten months old boy with obstruction of the ureterovesical junction and severe hydronephrosis, we discuss the validity of the assessment of renal function by radionuclide urography. Normal postoperative excretion as well as renographic controls confirmed the good relative function of the hydronephrotic kidney. Instead of nephrectomy a reconstructive operative procedure was employed. PMID- 8233848 TI - Misfortunes of war. PMID- 8233849 TI - Goodbye to all that? PMID- 8233850 TI - Taking control. PMID- 8233851 TI - Sticky wicket. PMID- 8233852 TI - Community chaos? PMID- 8233853 TI - Partners in care. PMID- 8233854 TI - Keeping a high profile. PMID- 8233855 TI - Continuing education. No easy option. PMID- 8233856 TI - Night school. Cystic fibrosis. PMID- 8233857 TI - Transplanting hope. PMID- 8233858 TI - Out in the open? Lesbianism. PMID- 8233859 TI - The experience of infectious patients in isolation. AB - Patients with infections are sometimes nursed in side rooms or isolation units. Little is known of how this experience of being isolated is perceived from either the patient's or the nurse's viewpoint. In this research eight patients and nurses were interviewed to explore their perceptions. Analysis indicated a wide range of response, with some patients valuing the privacy, solitude and control that the situation provided. However, expressions of neglect and isolation were common, and some patients perceived themselves to be stigmatised and lonely. Nurses described their patient's responses fairly accurately, but their ability to alter practice to meet the needs of these patients appeared constrained by time, the physical environment, fear of infection and the limitations of defining nursing interventions primarily in terms of physical care. PMID- 8233860 TI - Stroke care. Altered circumstances. PMID- 8233861 TI - Continence. A dry run. PMID- 8233862 TI - Continence. Picking and choosing. PMID- 8233863 TI - Employers urged to adopt HIV policy. PMID- 8233864 TI - A difficult age? PMID- 8233865 TI - A credibility problem. PMID- 8233866 TI - Home unrest. PMID- 8233867 TI - Learning quality. PMID- 8233868 TI - Preventing suicide. PMID- 8233869 TI - Suicide. Logical conclusion? PMID- 8233870 TI - Midwifery. Learning practice. PMID- 8233871 TI - Making sense of sex-linked inheritance. PMID- 8233872 TI - Writing a care study. PMID- 8233874 TI - Therapeutic touch. PMID- 8233873 TI - Professional development. Preparing for the future. PMID- 8233875 TI - Confronting prejudice. Gay and lesbian issues. PMID- 8233876 TI - Diabetes. Management training. PMID- 8233877 TI - Rehabilitation challenge. PMID- 8233878 TI - Positively passive. PMID- 8233879 TI - Power trip. PMID- 8233880 TI - A time for rebirth. PMID- 8233881 TI - A dangerous precedent. PMID- 8233882 TI - East bound. PMID- 8233884 TI - Mental health. Talking blues. PMID- 8233883 TI - Rites of passage. PMID- 8233885 TI - Maternal instincts. PMID- 8233886 TI - Absorbing effects. PMID- 8233887 TI - Treatment for life. PMID- 8233888 TI - War babies. PMID- 8233889 TI - Dignity under fire. PMID- 8233890 TI - Graduates only? PMID- 8233891 TI - Mental health. Learning independence. PMID- 8233892 TI - Wound care. Bridging the gap? PMID- 8233893 TI - Wound care. An appropriate choice. PMID- 8233894 TI - Teaching and learning in practice. 3. The teaching-learning relationship. (i) What is teaching? PMID- 8233895 TI - Aborted hopes. PMID- 8233896 TI - All mixed up. PMID- 8233897 TI - Reborn in the USA? PMID- 8233899 TI - Time to get changed. PMID- 8233898 TI - The numbers debate. PMID- 8233900 TI - Quality management work would improve nursing-home standards. PMID- 8233901 TI - HIV transmission ideas outdated. PMID- 8233902 TI - Head injury. Coming back to life. PMID- 8233903 TI - Follow the leader. PMID- 8233904 TI - Head injury. Safe transfer. PMID- 8233905 TI - HIV/AIDS--partners in need. PMID- 8233907 TI - Systems of life. The eye and vision. 4. PMID- 8233906 TI - Learning disabilities--paving the way. PMID- 8233908 TI - Long-stay care. A long-term view. PMID- 8233909 TI - Professional development. Blow your own trumpet. PMID- 8233910 TI - Promoting preceptorship. PMID- 8233912 TI - Theatre nursing. Role over? PMID- 8233911 TI - On the road. PMID- 8233913 TI - Theatre nursing. Chilling facts. PMID- 8233914 TI - The teaching-learning relationship (ii). What's in a title? PMID- 8233915 TI - Heading the league. PMID- 8233916 TI - Graduating in Gaza. PMID- 8233918 TI - On target? PMID- 8233917 TI - Betrayal of trust. PMID- 8233919 TI - The quest for quality. PMID- 8233920 TI - Don't dismiss exercise. PMID- 8233921 TI - Home truths. Nurses' homes. PMID- 8233922 TI - Boxed in. Nurses' homes. PMID- 8233923 TI - Making the grade. PMID- 8233924 TI - Midwifery. By mouth or muscle? PMID- 8233925 TI - Leaps of knowledge. PMID- 8233926 TI - Community nursing. A census of direction. PMID- 8233927 TI - London shake-up. A special case? PMID- 8233928 TI - Rights of way. PMID- 8233929 TI - Supporting dying people. AB - A descriptive interview survey was carried out to identify and assess the needs of dying people and their families and to see how far those needs were met by services available in the district. The main carers were interviewed six months after the death, and the majority said they were satisfied with care offered. However, a detailed analysis of comments made in response to the open-ended questions that were asked sheds light on the difficulties faced by many people in gaining access to services. PMID- 8233930 TI - Don't hold your breath. PMID- 8233931 TI - Wound care. Reducing the pressure. PMID- 8233932 TI - Wound care. Mattresses on trial. PMID- 8233934 TI - Wound care. Dressing practices. PMID- 8233933 TI - Wound care. A new policy of prevention. PMID- 8233935 TI - Healing alternatives. PMID- 8233936 TI - Teaching and learning in practice. 4. Rites, rights and power (i). Power and the individual. PMID- 8233937 TI - Cut to the quick. PMID- 8233938 TI - Morale dilemma. PMID- 8233939 TI - Cutting classes. PMID- 8233940 TI - Valuable assets. PMID- 8233941 TI - The next decade. PMID- 8233942 TI - Shocked by removal of cervix. PMID- 8233943 TI - Wielding the hatchet. PMID- 8233944 TI - Cot death. Reducing the risk. PMID- 8233945 TI - Cot death. Tragic statistics. PMID- 8233946 TI - Cot death. The never-ending loss. PMID- 8233948 TI - Solving the problem. PMID- 8233947 TI - Complementary medicine. Herbal medicine. PMID- 8233949 TI - The real Makoye. Interview by Daloni Carlisle. PMID- 8233950 TI - Birth pangs. PMID- 8233951 TI - Strong medicine. PMID- 8233952 TI - Stoma care--mixed meals. PMID- 8233953 TI - Mental health: sporting chances. PMID- 8233954 TI - Mental health: a change of perspective. PMID- 8233955 TI - Mental health: the cooperative dividend. PMID- 8233956 TI - Call to improve guidelines on antenatal HIV screening. PMID- 8233957 TI - Rites, rights and power (ii). The balance of power. PMID- 8233958 TI - Last rights. PMID- 8233960 TI - Body count. PMID- 8233959 TI - Open to abuse. PMID- 8233961 TI - Cervical screening. Testing times. PMID- 8233962 TI - Positive moves on HIV. PMID- 8233963 TI - Cervical screening. AB - Jane Hurley presents the results of a study that set out to establish women's attitudes towards a cervical screening programme and whether this affected their use of the service. The majority of women participating had used the service, but a clear need for improvement was identified. The need for more information about the purpose of cervical screening was highlighted, as was the need for health professionals to improve their communication skills. PMID- 8233964 TI - Learning disabilities. Fit for the future. PMID- 8233965 TI - Family crisis. PMID- 8233966 TI - Making sense of radio-frequency endometrial ablation. PMID- 8233967 TI - Off to a poor start? PMID- 8233968 TI - Ethnic minorities. Healthy relations. PMID- 8233969 TI - The kitchen's on fire. PMID- 8233970 TI - Speaking up. PMID- 8233971 TI - A&E nursing. Naming names. PMID- 8233972 TI - A shared vision. PMID- 8233973 TI - Theatre of memories. PMID- 8233974 TI - New group launched for HIV carers. PMID- 8233975 TI - UKCC changes AIDS guidelines for staff. PMID- 8233976 TI - Dietary fat and fiber modulate colonic cell proliferation in an interactive site specific manner. AB - A 3 x 3 factorial experiment was conducted to examine how dietary fiber and fat interactively affect cell proliferation in the colon in rats. Groups of 10 male Sprague-Dawley rats were fed one of nine experimental diets for three weeks: three types of fat at 15% by weight (beef tallow, corn oil, and fish oil) x 2 types of fiber (pectin and cellulose) plus a fiber-free diet as a control. Cell proliferation was measured by in vivo incorporation of bromodeoxyuridine into DNA. The main effect of dietary fiber on cell proliferation was found in the proximal colon, where the pectin diet stimulated cell proliferation compared with the cellulose and fiber-free diets (p < 0.05). In comparison, the main effect of dietary fat was in the distal colon, where the beef tallow diet was more promotive with respect to cell proliferation than the fish oil diet and the corn oil diet had an intermediate effect (p < 0.05). In the proximal colon, however, the effect of fiber on cell proliferation was highly dependent on the source of fat in the diet. Pectin exerted a hyperproliferative effect when the source of fat in the diet was corn oil, but not when beef tallow or fish oil was the fat source. These data indicate that dietary fiber and fat modulate cell proliferation of the colon in an interactive site-specific manner. PMID- 8233977 TI - Suppression of growth by dietary fish oil of human breast carcinomas maintained in three different strains of immune-deficient mice. AB - It has been reported that high levels of dietary fish (menhaden) oil, compared with corn oil, suppress the growth of MDA-MB231 and MCF-7 human breast carcinomas maintained in female athymic nude (T lymphocyte-deficient) mice. The purpose of this study was to determine whether dietary fish (menhaden) oil, compared with corn oil, can also suppress the growth of these carcinomas when maintained in female beige-XID-athymic nude (T lymphocyte- and NK/LAK cell-deficient) mice and in female severe combined immune-deficient (SCID) mice (total lack of functional T and B lymphocytes). Results clearly show that dietary fish (menhaden) oil can significantly (p < 0.05) suppress the growth of these carcinomas in the beige-XID athymic nude mouse and the SCID mouse. Such results provide evidence that the growth suppression of MDA-MB231 and MCF-7 human breast carcinomas, induced by dietary fish oil, is not mediated by immune system mechanisms involving T lymphocytes, B lymphocytes, and/or NK/LAK cells. PMID- 8233978 TI - Bioavailability of selenium from selenium-enriched garlic. AB - We previously reported that garlic grown in a selenium-fertilized medium (selenium-enriched garlic) is superior to regular garlic in mammary cancer prevention in an animal model (Nutr Cancer 17, 279-286, 1992). The present study was designed to evaluate the nutritional bioavailability of selenium from this garlic with use of two liver selenoenzymes as biomarkers: glutathione peroxidase and type I 5'-deiodinase. Rats were fed a selenium-deficient diet (0.01 ppm Se) from weaning for four weeks to deplete both enzymes. They were then supplemented with nutritional levels of selenium (0.1-0.5 ppm) in the form of sodium selenite (positive control) or selenium-enriched garlic. Our results showed that selenium enriched garlic was just as effective as selenite in restoring the activity of both selenoenzymes. This was demonstrated in a time course repletion experiment as well as in a dose-response experiment. Thus the selenium in selenium-enriched garlic has potent nutritional and anticancer efficacy. The type I 5'-deiodinase enzyme catalyzes the conversion of thyroxine (T4) to 3,5,3'-triiodothyronine (T3) and is responsible for most of the circulating T3. Because cancer chemoprevention by selenium usually requires pharmacological levels of selenium, we also examined the possible modulation of type I 5'-deiodinase by long-term feeding of selenium enriched garlic at 3 ppm Se in the diet. The observation that a high intake of selenium-enriched garlic did not affect 5'-deiodinase activity suggests that its anticarcinogenic effect is unlikely to be mediated by an imbalance in the blood T4-to-T3 ratio. PMID- 8233979 TI - Inhibition of oral carcinogenesis by glutathione. AB - Forty young adult Syrian hamsters (Mesocricetus auratus) were divided into four groups of 10 animals each. In Group 1 (tumor control), the right buccal pouches were painted three times per week with a 0.5% solution of 7,12 dimethylbenz[a]anthracene (DMBA) in heavy mineral oil (USP) with a no. 4 sable brush. In Group 2 (experimental group), the right buccal pouches were painted with DMBA, as in Group 1. In addition, Group 2 received 1 mg of reduced glutathione in 0.5 ml of mineral oil three times per week on days alternate to the DMBA application. The glutathione was administered systemically by mouth with a pipette. Group 3 received only glutathione, and Group 4 was untreated (control groups). Animals were sacrificed after 14 weeks, and tumors were counted and measured. Both right and left pouches were photographed, excised, fixed in formalin, sectioned in paraffin, and studied histologically. The animals receiving glutathione demonstrated significantly fewer and smaller tumors. The mean tumor burden was 315 mm3 in the glutathione-treated group and 3,040 mm3 in the untreated group. The statistical significance by Student's t test was < or = 0.0001. Histological study also revealed significantly fewer areas of dysplastic leukoplakia in the group treated with glutathione. This study represents the first demonstration of the anticancer effect of systemically administered reduced glutathione. PMID- 8233980 TI - The effectiveness of a mixture of beta-carotene, alpha-tocopherol, glutathione, and ascorbic acid for cancer prevention. AB - Previous studies have shown that beta-carotene and alpha-tocopherol can act synergistically to inhibit the growth of experimentally induced oral cancer. The initial studies on the synergistic anticancer activity of antioxidants have been extended to include reduced glutathione and ascorbic acid. Sixty male hamsters (4 5 wks old) were divided into six equal groups. Groups 1-6 were treated with 7,12 dimethylbenz[a]anthracene (DMBA) (0.5% solution). Group 2 received a mixture containing equal amounts of beta-carotene, dl-alpha-tocopherol (vitamin E), glutathione, and l-ascorbic acid (vitamin C) (12.5 micrograms) delivered orally by pipette. Groups 3-6 were treated with beta-carotene alone (50 micrograms), vitamin E alone (50 micrograms), glutathione (50 micrograms) alone, and vitamin C alone (50 micrograms). Animals were euthanized at 12 and 14 weeks. Tumors were counted and measured, and tumor burden was calculated for each experimental group. The mixture of antioxidants significantly reduced tumor burden, whereas the beta-carotene, vitamin E, and reduced glutathione treatments also reduced tumor burden. beta-Carotene and glutathione provided greater levels of chemoprevention than vitamin E as single agents. In contrast, vitamin C treatment produced no antitumor effect but increased tumor burden by Week 14. This mixture of antioxidants produced a significant synergistic chemoprevention of oral cancer. PMID- 8233981 TI - A comparison of diets of blacks and whites in three areas of the United States. AB - Dietary factors may contribute to the increased cancer risk of blacks. As a first step to explore this hypothesis, we examined food frequency data obtained by interview with 1,976 adults (881 blacks and 1,095 whites) randomly selected from three areas of the United States. The a priori hypothesis was that blacks were more likely to consume diets low in fruits and vegetables and/or high in fat, particularly saturated fat. Contrary to expectation, blacks were more frequent consumers of fruits and vegetables considered to be protective against cancer (e.g., citrus fruits, cruciferous vegetables, and vegetables rich in vitamins A and C). Intake of both total and saturated fat was slightly lower among blacks than whites. This analysis does not rule out a role for these dietary factors in the etiology of cancer but indicates that ascribing the excess cancer risk among blacks to their frequency of fruit and vegetable consumption or intake of fat per se is inadequate. This suggests that alternative dietary explanations for the racial disparity in cancer risk should be pursued in future studies. PMID- 8233982 TI - Dietary factors and survival from breast cancer. AB - The association between self-reported intake of various dietary factors at diagnosis and survival from breast cancer was studied in a population-based cohort of breast cancer patients in Adelaide, South Australia. These patients had been recruited between 1982 and 1984 into a case-control study of diet and incident breast cancer. Of the 451 patients recruited originally, 412 were followed for a median interval of 5.5 years. There were decreases in the risk of death from breast cancer ranging from 25 to 40% at all levels of energy and protein intake above the baseline, whereas for fat intake there was a 40% increase in risk at the uppermost quintile level. There was also some reduction in risk at the upper levels of intake of beta-carotene and vitamin C. However, there were no dose-dependent variations in risk of death by level of intake for any of the dietary factors studied, and most of the variation in risk that was observed was relatively insubstantial. PMID- 8233983 TI - Relation of body size and the risk of lung cancer. AB - Two population-based case-control studies of lung cancer were conducted on the island of Oahu, HI, between 1979 and 1985. Data from these studies were combined to form the basis of this analysis. Interviews were obtained from 518 men and 230 women with lung cancer and 1,102 male and 524 female controls frequency matched to the cases by sex and five-year age group. The interview consisted of a complete tobacco history, information on body size, and other demographic and life-style information. Weight and Quetelet index (kg/m2) 5 years before diagnosis, but not at 20-29 years of age, were inversely related to the risk of lung cancer among men and women. Cases tended to gain less weight during adulthood than did controls. These associations persisted after adjustment for age, ethnicity, tobacco smoking history, and beta-carotene intake. Our results are consistent with reports by several other investigators of an inverse association between body weight and the risk of lung cancer. However, we were unable to rule out the possibility of bias in our findings due to preclinical disease. PMID- 8233984 TI - Dietary restriction reduces the incidence of NMU-induced mammary tumors and alters retinoid tissue concentrations in rats. AB - Previous studies suggested a relationship between dietary restriction (DR) effects on mammary carcinogenesis and DR effects on liver retinoids. Therefore, in this study, retinoid concentrations were measured by high-performance liquid chromatography in the plasma, liver, and peripheral organs of DR rats with chemically induced carcinogenesis. Rats were injected with N-methyl-N-nitrosourea (MNU) and maintained on graded levels of DR (reduction of 10-40% from energy ingested by control animals with free access to food). Mammary tumor incidence and multiplicity induced by MNU were reduced in relation to the degree of DR, with virtual prevention occurring at 30% and 40% DR. Total hepatic retinoid concentrations (retinol + retinyl esters) were significantly greater in rats given MNU and subjected to DR, but liver total retinoid content was comparable between the groups. However, plasma retinol concentrations were significantly lower in DR rats than in controls given the carcinogen without DR. Retinoid concentrations were also elevated in adipose tissue, lungs, and intestine of DR rats, while renal concentrations remained unaltered. Retinoid concentrations in mammary glands and mammary tumors were similar in all groups. Thus, in DR rats, vitamin A concentrations in liver and other target tissues are maintained or increased despite decreases in plasma. It remains to be investigated whether these alterations in retinoid content have any relationship to the cancer preventive effect of DR. PMID- 8233985 TI - Menhaden, coconut, and corn oils and mammary tumor incidence in BALB/c virgin female mice treated with DMBA. AB - Omega-3 fatty (n-3) acids are believed to inhibit the rate of occurrence and the growth of mammary tumors in rats treated with 7,12-dimethylbenz[a]anthracene (DMBA). Linoleic acid, on the other hand, has been shown to promote mammary tumorigenesis. This study was undertaken to see whether replacing 18% of the corn oil (high in linoleic acid) in a 20% fat diet with menhaden oil (high in n-3 fatty acids, low in linoleic acid) or coconut oil (low in n-3 fatty acids, low in linoleic acid), while keeping constant the cholesterol, antioxidant, and total fat content, would affect tumor incidence in virgin female BALB/c mice dosed with DMBA. Dietary treatment had no effect on body weight, feed intake, or survival to 44 weeks of age (36 wks after the first of 6 DMBA doses). Mammary tumor incidence was the same in the menhaden oil and coconut oil diet groups but was significantly higher in the 20% corn oil diet group. The protective effect of menhaden oil and coconut oil may be due, at least in part, to the decreased linoleic acid content of these diets relative to the corn oil diet. We conclude that n-3 fatty acids per se do not seem to inhibit tumor formation. PMID- 8233986 TI - Dental attitudes and memories: a study of the effects of hand over mouth/restraint. AB - The purpose of this study was to document whether there was a significant difference in the number and severity of generalized fears and dental fears between patients who did and patients who did not experience hand-over-mouth and/or restraint as children. Patient records in a dental school children's clinic and a private pediatric dental practice were examined to identify patients who had experienced hand-over-mouth and/or restraint. A set of verbal questions was designed, tested, and used to ascertain the differences between the HOM/restraint group and the comparison group. One hundred twenty-two subjects were interviewed, 61 who had experienced HOM/restraint and 61 who had not. When compared for generalized fears and specific dental fears, the two groups showed no statistically significant differences (P = 0.86 and P = 0.36 respectively). No statistically significant difference appeared between the two groups when asked how they felt about visiting the dental office (P = 0.41). When three different formats were used to question the subjects relative to their early dental memories, the two groups showed no statistical difference in negative or positive responses (P = 0.38, 0.75, and 0.25 respectively). More than two times as many HOM/restraint subjects as comparison subjects described negative experiences in a physician's office or hospital. This difference was statistically significant (P < 0.01). PMID- 8233987 TI - Dentist's reassuring touch: effects on children's behavior. AB - Physical contact with patients by health care providers has been found to benefit the patients by reducing their fearful or avoidant reactions. This study tested whether a reassuring touch could be used during a routine pediatric dental examination to reduce children's anxiety and improve their behavior. Thirty-eight children between 3.5 and 10 years of age were randomly assigned to one of two experimental conditions. Children assigned to the touch condition were patted on the upper arm or shoulder on two separate occasions by the dentist during the examination while simultaneously receiving verbal reassurance and descriptions of the upcoming procedures. Children in the no-touch control condition received only the reassuring verbal descriptions without contact. Results indicated that touched children between the ages of 7 and 10 years (but not children aged 3.5 to 7 years) displayed less fidgeting behavior than their no-touch counterparts (P < 0.05). Post-treatment, children who were touched tended to report greater pleasure (P < 0.06) but less dominance (P < 0.10) than children not touched. PMID- 8233988 TI - Variables influencing hemoglobin oxygen desaturation in children during routine restorative dentistry. AB - Pulse oximetry to monitor oxygen saturation during pediatric dental sedations has revolutionized the early detection of hypoxemia. Previous sedation studies confirmed the occurrence of hemoglobin oxygen desaturation when sedative agents are administered in conjunction with nitrous oxide. The purpose of this study was to monitor the hemoglobin oxygen saturation levels of nonmedicated pediatric patients during routine restorative procedures to study the effect of nitrous oxide, chronologic age, tonsil size, and lidocaine dose. Two identical pulse oximeters and probes were attached to a patient's right and left feet to validate the occurrence of oxygen desaturations. The results revealed that 20.3% of the patients demonstrated at least one occurrence of oxygen desaturation during the routine dental procedure. Administration of nitrous oxide had no effect on the prevalence of oxygen desaturation. Although chronologic age, tonsil size, and lidocaine dose did not exert a statistically significant influence when analyzed independently, when combined they were related statistically (P < 0.0001) to the occurrence of oxygen desaturation as defined by the parameters of the study. These data suggest that some of the oxygen desaturations recorded during routine restorative appointments may result from probe error and a combination of patient related variables. PMID- 8233989 TI - Delayed tooth formation in low birthweight African-American children. AB - Low birthweight (LBW) infants are susceptible to several developmental problems (e.g., pulmonary diseases, hyperbilirubinemia, hypocalcemia) with potentially long-lasting effects that slow growth during infancy and childhood. Dental age (DA), judged from stages of permanent tooth mineralization, was scored in 4- to 7 year-old LBW African-American children (N = 66; x = 5.5 years) to test whether they were delayed due to LBW and its consequences. Data were matched in a case control fashion to African-American children with normal birth-weights (N = 76). Only the early-forming teeth (incisors, first molars) were delayed significantly in their formation. Children with the lowest height-for-age centiles were the most delayed dentally which suggests that height status would improve as dental age caught up with chronological age (CA). Older children were more delayed because there is a proportionately greater opportunity for DA to diverge from CA as children grow older. Since only those teeth undergoing rapid differentiation neonatally were affected systematically, it was speculated that perinatal insults may have an enduring impact on developing primordia, while leaving later-forming teeth unaffected. PMID- 8233990 TI - Project USAP the use of sedative agents in pediatric dentistry: 1991 update. AB - A national survey of members of the American Academy of Pediatric Dentistry was conducted to update information on the use of sedative agents by pediatric dentists. All 2532 members of the Academy were sent questionnaires regarding their use of sedation and 1497 responded. Practitioners were questioned regarding how frequently they used sedative agents and the nature of patients receiving sedation. In addition, they were questioned on reasons for change, if any, in their use of sedation during the past two years. A separate survey was conducted of directors of all 55 postdoctoral training programs in pediatric dentistry. Forty-six directors responded to questions concerning the type and frequency of sedation use in their programs. Both surveys demonstrated wide differences in sedation use between different training programs and between different practitioners. The heavier use of sedation by some practitioners or by some training programs, as compared with others, generally was not related to the percentage of handicapped patients who received sedation, nor to the type of training. Frequent users of sedation (more than once each day) were located more in the south/southeastern and western parts of the country and they tended to have been in specialty practice for more than 10 years. PMID- 8233991 TI - Social and biological factors contributing to caries of the maxillary anterior teeth. AB - Caries prevalence among 3- to 4-year-old Head Start children and psychosocial information from their parents were obtained. The prevalence of maxillary anterior caries in the 369 children was 16%, with the most severely affected tooth surfaces being the mesial surfaces of the central incisors. Approximately 90% of parents whose children were found to have maxillary anterior caries knew that allowing their child to take a bottle to bed would harm the child's teeth. Significant differences were found in mutans streptococci levels between children with and without the anterior caries pattern. Although 86% of children with anterior caries were reported to have taken a bottle to bed, 69% of those children who did not have anterior caries also were reported to have taken a bottle to bed. Of the children with maxillary anterior caries, 87% had posterior caries, suggesting that anterior caries subsequently may contribute to an increased caries risk in other teeth. PMID- 8233992 TI - Avulsion of a maxillary primary first molar in a 19-month-old child. PMID- 8233994 TI - American Board of Pediatric Dentistry Clinical Section: Case Review Option. Guidelines and instructions for candidates. Approved September 1991. PMID- 8233993 TI - Spondylometaphyseal dysplasia (Kozlowski type): case report. AB - Kozlowski syndrome is the most common type of spondylometaphyseal dysplasia (SMD). It is characterized by short stature (130 to 150 cm), pectus carinatum, limited elbow and hip movement, mild bowleg deformity, and curvature of the spinal column. Children with Kozlowski dwarfism usually are not recognized at birth, since they have normal clinical features, weight, and size. This article reports the dental treatment and oral findings of a 14-year-old female patient with Kozlowski dwarfism. PMID- 8233995 TI - Chemoradiation therapy: effect on dental development. AB - Chemoradiation therapy used on pediatric oncology patients often causes dental developmental anomalies that affect future dental care. Defects noted include tooth and root agenesis, root thinning and shortening, and localized enamel defects. Histologically, these defects appear as osteoid-like niches in the developing dentin which alter the overlying enamel. Odontogenic cell sensitivity is dependent upon the position on the cell cycle and the mitotic activity at the time of chemoradiation therapy. Knowledge of the stage of dental development at the time of oncology treatment and the type of therapy allows the clinician to predict dental effects of the chemoradiation. Representative cases illustrate the clinical manifestations of chemoradiation on the developing dentition. PMID- 8233996 TI - Pathophysiology of heart failure. AB - Heart failure, a major contributor to cardiovascular disease morbidity and mortality, is newly diagnosed in approximately 400,000 patients each year, and is particularly prevalent in individuals over age 65 years. Average mortality rates 5 years after diagnosis are 45-60%, and may be as high as 50% after 1 year for those with New York Heart Association class IV heart disease. Heart failure occurs when myocardial muscle dysfunction prevents the heart from pumping enough blood at normal cardiac pressures to meet the metabolic needs of the body, especially during exercise, and compensatory hemodynamic and neurohormonal mechanisms are overwhelmed or maladaptive. Pathologic classifications are broadly based on the presence of systolic (dilated cardiomyopathy) or diastolic (hypertrophic or restrictive cardiomyopathies) dysfunction. The etiologies of heart failure may include inadequate coronary blood flow, pressure or volume overload, cardiomyopathy, or pericardial disease. Coronary artery disease, idiopathic dilated cardiomyopathy, and hypertension are the most frequent causes, and certain drugs may also worsen myocardial function. When contractility is reduced, stroke volume and cardiac output are decreased, and alterations in the kidneys may induce fluid retention to compensate for the perceived low output and reduced circulating blood volume. Fluid retention in turn causes preload or filling pressure to increase and symptoms of pulmonary congestion to emerge. Depressed contractility also results in a reduction in blood pressure, leading to compensatory neurohormonal activation and vasoconstriction, which significantly elevate afterload and further reduce stroke volume. The overall approach to heart failure includes defining the etiology, identifying precipitant factors, and assessing the severity of myocardial dysfunction and clinical symptoms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233997 TI - Assessing the treatment of congestive heart failure: diuretics, vasodilators, and angiotensin-converting enzyme inhibitors. AB - Congestive heart failure (CHF) causes disabling symptoms and increases the likelihood of decreased survival. Diuretics, direct vasodilators, and angiotensin converting enzyme (ACE) inhibitors can be used to reduce symptoms, prolong life, or both, in these individuals. Diuretics induce sodium and water excretion, leading to decreased cardiac preload and wall tension, and an effective decrease of symptomatic pulmonary and systemic congestion. They have not yet been shown to prolong life in patients with CHF, however. Direct vasodilators, which induce venodilation, arterial dilation, or both (balanced vasodilators), may improve symptoms, and some but not all prolong life. Venodilators, such as nitrates, exert a venous pooling effect, decreasing cardiac preload and symptoms of congestion. Arterial dilators, such as hydralazine, decrease afterload and improve cardiac output. The combination of hydralazine and isosorbide dinitrate provides balanced vasodilation. It also improves survival, but is associated with a relatively high frequency of side effects necessitating discontinuation of one or both agents. The drugs are not approved by the Food and Drug Administration for the treatment of heart failure. Flosequinan, a new orally administered, long acting, balanced arteriovenous dilator, improves exercise tolerance and symptoms. However, preliminary analysis of data from a large, multicenter trial revealed increased mortality and hospitalization for worsening CHF. The drug has recently been withdrawn from the market. The ACE inhibitors can cause hemodynamic and neurohormonal changes that lead to a reduction of preload and afterload, decreasing symptoms of heart failure. They significantly decrease CHF mortality, and might also deter the development of overt heart failure in some asymptomatic patients with left ventricular dysfunction. PMID- 8233998 TI - Clinical trial strategies and future challenges in the investigation of human congestive heart failure. AB - Clinical research in the therapeutics of chronic congestive heart failure (CHF) has evolved from simple case reports and general-response studies in the 1960s to large, randomized, controlled trials conducted in the late 1980s and 1990s. Cumulatively, the investigations show that angiotensin-converting enzyme (ACE) inhibitors improve clinical status, exercise capacity, and survival; digoxin augments exercise capacity and, probably, clinical status, with unknown effects on survival; the vasodilator combination hydralazine-nitrate improves survival, and perhaps clinical status and exercise capacity; and the new vasodilator flosequinan has a favorable effect on clinical status and exercise capacity, but is associated with an increased risk of mortality at 100 mg/day (this agent has subsequently been withdrawn voluntarily by the manufacturer from clinical use). Advances in study design and methodology, combined with the outcomes of major clinical trials, have resulted in new challenges in the investigation of CHF in humans. To date, only the ACE inhibitors have been shown to affect all of the major CHF therapeutic end points in a positive manner. Is it reasonable to expect similar results from all future drug interventions? If not, which end points should be targeted? Must the survival end point be investigated for all CHF drugs under development? Can studies be performed ethically without background ACE inhibitor therapy? Have the optimum duration and methods for therapeutic trials in humans been determined? Should controlled trials of nonhemodynamic and nonneurohormonal interventions (e.g., anticoagulation, magnesium, exercise regimens) be performed? How can the results of clinical trials best be applied in treating the broad range of patients encountered in practice?(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8233999 TI - [Bone marrow transplantation--present status and perspectives]. PMID- 8234000 TI - [Influence of progressive systemic sclerosis on gastric emptying]. AB - Gastric motility was measured with an isotopic method in patients with progressive systemic sclerosis (PSS). Twenty-six women aged 48 +/- 10 yr with PSS were investigated. The duration of PSS prior to the study ranged from 2 to 25 yr. Control values were obtained from 12 healthy female volunteers aged 34 +/- 8 yr. Endoscopic examination was carried out in all investigated individuals to exclude abnormalities that were non-related to PSS. Measurements were done at morning, after an overnight fast. Patients and controls received a test meal of 1650 kJ containing human albumin microspheres labelled with 7.4 MBq of 99mTc. Subjects were investigated with a gamma-camera. It was shown that gastric emptying (GE) t 1/2 was delayed in the patient with PSS (289 +/- 325 min) as compared to the controls (92 +/- 19 min), p < 0.001. Delayed t 1/2 over x + 2SD of the control group was found in 69% of patients with PSS while in the control group O. The mean transit time (MTT90) was in the PSS group significantly longer than in the control group, 42.3 +/- 1.4 and 40 +/- 0.9 min respectively (p < 0.001). There was no correlation found between GE t 1/2 and the time of PSS duration (r = 0.08), its type II or III as well as the titer of Scl-70 antibodies in blood serum (r = 0.29, p > 0.1). PMID- 8234001 TI - [Level of beta endorphins and insulin in blood of obese subjects. Effect of surgical treatment for obesity on higher exchange parameters]. AB - The basal and hyperglycaemia-stimulated secretion of glucose, IRI and beta endorphin (BE) were studied in subjects who had gone surgical treatment for obesity few years ago and the results were compared with those of obese subjects and lean controls. 58 persons were divided into the following groups: A-obese subjects BMI > 30, B--obese subjects 25 < BMI < 30, C--subjects treated by truncal vagotomy and gastric banding, D--subjects treated by jejunoileostomy, E- control group BMI < 25. Oral glucose (75 g) tolerance test was performed in all subjects. Blood concentration of glucose, and serum concentration of IRI and BE were studied before and 30, 60, 90 and 120 minutes after ingestion of glucose. The basal levels and areas over basal values (AOBV) of investigated parameters were evaluated. Both the basal and glucose stimulated levels of IRI and BE were higher in the obese subjects than in the control group. Truncal vagotomy and gastric banding or jejunoileostomy resulted in reduction of IRI secretion without any decrease in BE levels. The alteration of the opioid system may play some role in the pathogenesis of obesity. PMID- 8234002 TI - [Evaluation of uric acid transport in nephrons of patients with calcium nephrolithiasis and hyperuricosuria]. AB - Renal excretion of uric acid was studied in 48 patients with recurrent calcium nephrolithiasis. Hyperuricosuria was found in 16 patients (33%) and among them 10 patients had increased values of urate clearance and fractional excretion. The tubular transport of urate was evaluated by means of pharmacological tests with pyrazinamide (PZA) and benzbromarone (BB) in these 11 patients with hyperuricosuria of renal origin. PZA suppression test was normal in 10 patients while in one case uric acid excretion was not suppressed sufficiently by PZA. Maximal uricorusirc response to BB was increased in two patients, normal in one patient and impaired in eight patients. PZA and BB tests revealed isolated defect of uric acid postsecretory reabsorption in seven patients, impaired reabsorption at both pre- and postsecretory site in one case and enhanced tubular secretion of uric acid in two patients. This study indicates that different defects of uric acid transport in nephron are the frequent cause of hyperuricosuria in patients with recurrent calcium nephrolithiasis. PMID- 8234003 TI - [Measurement of glycosylated hemoglobin as a useful method for controlling type II diabetes mellitus in patients suspected of incomplete compensation]. AB - The conduction of levels of glycosylated hemoglobin in patients with type II diabetes mellitus was studied A group of 111 ambulant patients was analyzed and special attention paid to those patients who were given the highest permissible oral dose. The dependence between the achieved glycosylation tests results and types of therapy, the clinical course of diabetes mellitus, as well as the conduction of results of standard compensation tests was analyzed. A lower correlation degree between the level of HbA1 and the results of standard compensation tests was indicated. At the same time a high correlation degree between HbA1 and clinically proven diabetes complication progress was observed. All achieved results suggest the usefulness of HbA1 determination in patients with type II diabetes mellitus suspected of incomplete compensation for instance treated highest permissible oral dose. PMID- 8234004 TI - [Reactive hyperinsulinemia in coronary artery disease. 1]. AB - Two groups of patients were studied: group with stable coronary artery disease and group without the coronary artery disease. Classical risk factors of coronary artery disease as well as glycaemia and insulinemia during the oral glucose tolerance test was investigated. Positive correlation between coronary artery disease and classical risk factors as well as reactive insulin level was found with normal reactive glycemia. PMID- 8234005 TI - [Reactive hyperinsulinemia in coronary artery disease. 2. Correlation between reactive hyperinsulinemia and other risk factors of coronary artery disease]. AB - Two groups of patients were studied: group with stable coronary artery disease and group without the coronary artery disease as well as glycaemia and insulinaemia were studied during the oral glucose tolerance test. The results show positive correlation between hyperinsulinemia and some classical risk factors of coronary artery disease. PMID- 8234006 TI - [Liver in Wilson's disease]. PMID- 8234007 TI - [Glucose-6-phosphate dehydrogenase deficiency in erythrocytes observed in two cases]. AB - In two male patients the deficiency of G-6-PD in erythrocytes became manifested in 27th year of age with severe hemolytic crisis. In the first case hemolysis appeared after three days of heavy alcohol drinking. There was no evident relation between alcohol and hemolysis. In the second patient (an Iraqi Student) suffering from diabetes since adolescence, hemolysis was provoked by respiratory tract infection, antiinflammatory drugs and concomitant ketosis. Pneumonia, renal insufficiency and acute pancreatitis complicated the course of illness. The patient required a consecutive erythrocyte concentrate transfusions. Aplastic crisis was treated with folic acid. PMID- 8234008 TI - [Eleven year period of asymptomatic HIV infection in a patient after open heart surgery]. AB - A case is described of a 62 years old patient in the initial stage of clinically overt HIV infection. The infection occurred probably 11 years earlier by means of blood transfusion during open heart surgery. The possible reasons for the long term asymptomatic carriage of the virus and factors influencing the development of symptomatic HIV infection (AIDS) are discussed. PMID- 8234009 TI - Structure-activity relationships for inhibitory insect myosuppressins: contrast with the stimulatory sulfakinins. AB - Unusual among insect neuropeptides, the decapeptide myosuppressins are capable of inhibiting contractions of visceral muscle, including the isolated cockroach hindgut. The C-terminal pentapeptide Val-Phe-Leu-Arg-Phe-NH2 has been identified as the myosuppressin active core, the minimum number of residues required to elicit hindgut myoinhibitory activity. Activity of the same magnitude as the parent neuropeptide requires the C-terminal heptapeptide fragment Asp-His-Val-Phe Leu-Arg-Phe-NH2. Evaluation of a series of substitution analogs delineates structural features critical for myoinhibitory activity within this important fragment. The branched, hydrophobic residues in myosuppressin position 6 (Val) and particularly position 8 (Leu), their absence in the myostimulatory sulfakinins, and the different roles played by the shared Asp residue (myosuppressin position 4; leucosulfakinin position 5) in peptide-receptor interaction, account in large degree for the contrasting biological activities elicited by these otherwise structurally similar peptide families. The results may have broad significance for other invertebrate myotropic systems, such as the locust heart and the pharyngeal retractor muscle of the mollusc Helix aspersa. PMID- 8234010 TI - Cardiovascular actions of angiotensin(1-7). AB - Angiotensin(1-7) had a compound effect on blood pressure of pithed Sprague-Dawley rats. The initial phase of the response consisted of an increase in MAP of short duration and independent of injected dose, followed by a decline of arterial pressure to values below baseline. Both the magnitude (range: -4 +/- 1 to -13 +/- 1 mmHg) and the duration (range: 83 +/- 13 to 255 +/- 17 s) of the depressor response correlated with the dose of peptide. Indomethacin (5 mg/kg) eliminated the depressor component. Only [Sar1,Thr8]Ang II inhibited the effect of Ang(1-7) completely. We conclude that angiotensin(1-7) possesses myotonic actions that are in part related to release of vasodilator prostaglandins through an angiotensin receptor other than AT1 or AT2. PMID- 8234011 TI - Isolation, purification, and characterization of calcitonin gene-related peptide receptor. AB - Intact calcitonin gene-related peptide (CGRP) receptors were solubilized from porcine neural membranes using sodium cholate: potassium buffer. The solubilized receptors were purified sequentially by hydrophobic interaction and ion-exchange chromatography followed by specific affinity chromatography. Using these procedures, we have isolated 2 nmol of highly purified active CGRP receptor to a homogeneity (5 x 10(8)-fold purification). The isolated receptors retained their specificity and the capacity to bind to 125I-CGRP, and showed no cross-reactivity with a number of other peptides, except with amylin having 46% amino acid sequence homology to h-CGRP. The solubilized receptors were adsorbed by WGA agarose and concanavalin-A, suggesting a glycoprotein nature. SDS-PAGE, size exclusion HPLC, and autoradiography confirmed that CGRP receptor is a monomeric membrane protein with M(r) 66 kDa. PMID- 8234012 TI - Arginine vasopressin- and oxytocin-like peptides in the testis of two Australian marsupials. AB - High performance liquid chromatography (HPLC) and specific radioimmunoassay (RIA) for arginine vasopressin (AVP), mesotocin (MT), and oxytocin (OT) were used to identify and quantify these peptides in the testis of the brushtail possum (Trichosurus vulpecula) and the northern brown bandicoot (Isoodon macrourus). Arginine vasopressin (0.092 +/- 0.041 ng/g) and MT (0.198 +/- 0.089 ng/g), but not OT, were found in the possum testis, while the bandicoot testis contained AVP (0.061 ng/g), MT (0.108 +/- 0.024 ng/g), and OT (0.114 +/- 0.053 ng/g). The values correlate well with those reported for AVP- and OT-like peptides in the testis of eutherian mammals. It was concluded that there are neurohypophysial peptides present in the marsupial testis. PMID- 8234013 TI - Influence of growth hormone (GH) and GH-releasing factor on locomotor activity in rats. AB - Acute recombinant human growth hormone (r-hGH) administration (1-250 micrograms/kg; IP) decreased locomotor activity (LA) in a dose-dependent manner, F(5, 43) = 3.55, p < 0.009, with maximum effect at a dose of 10 micrograms/kg [control = 1655 +/- 659 inputs/outputs (I/O); r-hGH = 909 +/- 436 I/O; p < 0.05]. The daily treatment with r-hGH (10 micrograms/kg/day; 5 days; IP) reduced LA counts during the first trial, with no apparent changes in motor behavior after habituation. In contrast, growth hormone-releasing factor (GRF; 10 micrograms/rat/day; 5 days; IP) enhanced LA (control = 1045 +/- 566 I/O; GRF = 2284 +/- 894 I/O; p < 0.01) during the 5-day treatment period, inhibiting habituation. Moreover, the individual differences in LA persisted during the treatment period in response to GRF or r-hGH, but not in control rats. These results seem to indicate that the effects of GRF on LA are not mediated by the release of peripheral GH, and suggest that GRF might influence psychomotor behavior by a central mechanism. PMID- 8234014 TI - The ovine hypothalamus and pituitary have markedly different distribution of C type natriuretic peptide forms. AB - C-type natriuretic peptide (CNP) was measured in the hypothalamus and pituitary of four sheep by radioimmunoassay after extraction. The mean concentration of CNP in the hypothalamus was 1.01 +/- 0.08 pmol/g and 45.8 +/- 12.8 pmol/g in the pituitary. Analysis of these extracts by size exclusion HPLC showed the presence of two immunoreactive CNP components that cochromatographed with porcine CNP-53 and CNP-22 standards. Similar amounts of CNP-53- and CNP-22-like IR-CNP were present in the ovine hypothalamus (ratio 0.9:1), whereas in the pituitary, the bulk of the immunoreactive CNP was in the CNP-53-like form. These results show major differences in the distribution of IR-CNP forms between the hypothalamus and pituitary, which may reflect differences in CNP prohormone processing in these two tissues. PMID- 8234015 TI - Mechanism of bombesin-induced pancreatic secretion in unanesthetized rats. AB - It is unclear whether stimulation of pancreatic enzyme secretion by intravenously administered bombesin is a direct effect on acinar cells or is mediated by release of CCK; this distinction is important for defining the potential role of bombesin-like peptides as regulators of pancreatic secretion. The role of CCK in bombesin-induced pancreatic secretion was examined in rats using CCK radioimmunoassay and the CCK receptor antagonist L-364,718. A biphasic pancreatic response occurred to sequential doubling doses of bombesin (31 to 2000 pmol/kg/h, each for 30 min; n = 9 rats); amylase secretion increased to peak at 250 pmol/kg/h (11.5 +/- 1.7 kU/30 min; 4.2 +/- 0.6 kU/30 min, basal) and then declined to basal levels at 2000 pmol/kg/h. The ED50 dose of bombesin for stimulation was 31 pmol/kg/h, and the maximal response did not differ significantly from that to exogenous CCK-8 (10.6 +/- 1.5 kU/30 min) in the same rats. When single doses of bombesin were infused for 2 h (31, 62, 125, 250 pmol/kg/h; one dose per day; order randomized; n = 8), a similar dose-response relationship was seen, both for peak amylase response and cumulative output over basal. L-364,718 (0.5 mg/kg IV) had no effect on the pancreatic response to ED50 or maximal doses of bombesin. Neither dose of bombesin altered plasma CCK levels. In contrast, other stimulants of pancreatic secretion (food ingestion, soybean trypsin inhibitor) caused marked elevations in plasma CCK levels. These results indicate that the potent stimulation of pancreatic secretion by exogenous bombesin in rats is not mediated by CCK, similar to findings in humans. PMID- 8234016 TI - Multiple molecular forms of gonadotropin-releasing hormone in the brain of an elasmobranch: evidence for IR-lamprey GnRH. AB - These studies investigated brains of skate, Raja erinacea (order Rajiformes, class Chondrichthyes), for gonadotropin-releasing hormone (GnRH) peptides by chromatograph and immunoreactivity with region-specific antisera raised against mammalian GnRH and lamprey GnRH. The region-specific antibody to lamprey GnRH-I was produced following conjugation to bovine serum albumin using the bis diazotized benzidine method. This antibody was characterized by assaying a range of increasing dilutions of the known vertebrate GnRHs, as well as analogs to lamprey GnRH-I. Two analogs, lamprey [Phe2]GnRH-I and lamprey [Leu7]GnRH-I, were synthesized by solid phase peptide synthesis using a benzhydrylamine resin as the supporting medium and purified by chromatography. This antibody demonstrated less than 0.01% cross-reactivity with all GnRH peptides tested, suggesting a highly specific antibody with a region of amino acids 2-8 that appears essential for binding. In the skate brain, five immunoreactive (IR) GnRH forms were detected, four of which eluted in the same positions as synthetic mammal and chicken GnRH-I (which coelute): lamprey GnRH-I, salmon and chicken GnRH-II, and one that was an unidentified form. A minor peak coeluted with lamprey GnRH-III. The major form in the skate brain is considered to have eluted with synthetic mammalian GnRH. These studies confirm an earlier report of an IR-mammalian GnRH peptide and provide new evidence for IR-lamprey GnRH in the brain of an elasmobranch. PMID- 8234017 TI - Enhanced antiopiate activity and enzyme resistance in peptidomimetics of FMRFamide containing (E)-2,3-methanomethionine. AB - FMRFamide is a molluscan peptide that has shown antiopiate activity in a number of mammalian test systems. The current study determined the antiopiate potency of FMRFamide and two conformationally constrained peptidomimetics of FMRFamide containing stereoisomers of (E)-2,3-methanomethionine. Morphine abstinence signs were observed after varying doses (0.25-25.0 microgram) of these substances were injected into the third ventricle of morphine-dependent rats. Both peptidomimetics were far more potent than FMRFamide itself. In addition, although both peptidomimetics bound with lower affinity than FMRFamide to rat spinal cord receptors for NPFF (the mammalian FMRFamide-like peptide), they were far more resistant than FMRFamide to enzymatic degradation by leucine aminopeptidase. PMID- 8234018 TI - Crustacean cardioactive peptide in the sphinx moth, Manduca sexta. AB - The isolation, identification, and actions of crustacean cardiactive peptide (CCAP) have been examined in the sphinx moth Manduca sexta. A sensitive and specific enzyme-linked immunosorbent assay (ELISA) was used to quantify CCAP-like immunoreactivity in the nervous system. The CCAP-like immunoreactivity from the abdominal CNS was then purified, and its sequence was ascertained by amino acid analysis, mass spectral analysis, and HPLC. These studies showed that the nervous system of M. sexta contains a peptide with the sequence Pro-Phe-Cys-Asn-Ala-Phe Thr-Gly-Cys-NH2, identical to CCAP originally isolated and sequenced from the shore crab Carcinus maenas. The actions of CCAP on the isolated heart of M. sexta and the extensor-tibia muscle of Schistocerca americana were tested. Crustacean cardioactive peptide had excitatory actions on both preparations: a dose dependent increase in the rate of contractions was observed on the heart, and an increase in the rate of the myogenic rhythm was observed in the leg muscle. Moreover, purified and synthetic CCAP had identical effects on the isolated heart. We conclude that CCAP occurs in M. sexta and exerts potent neurotransmitter or neurohormonal actions on a variety of muscles. PMID- 8234019 TI - CRF in the paraventricular nucleus of the hypothalamus stimulates colonic motor activity in fasted rats. AB - The influence of corticotropin releasing factor (CRF) microinjected into the paraventricular nucleus of the hypothalamus (PVN) on colonic motility was investigated in conscious, fasted rats. Rats were chronically implanted with a bilateral guide cannula into the PVN and a catheter into the proximal colon to record motor activity manometrically. Microinjection of CRF (0.6 nmol/rat) into the PVN increased both phasic and tonic motor activity in the proximal colon. Atropine sulfate (1 mg/kg, IP) completely abolished the colonic motor response to CRF. Microinjection of CRF (0.6 nmol/rat) into sites outside of the PVN did not modify colonic motor activity. These data show that CRF acts in the PVN to stimulate tonic and phasic motor activity in the proximal colon. Corticotropin releasing factor action is site specific and mediated through cholinergic pathways. PMID- 8234020 TI - Effects of F8Famide analogs on intestinal transit in mice. AB - In an attempt to establish the role of F8Famide in opioid activity modulation, we examined the effects of intracerebroventricular administration of the F8Famide analogs (1DME)Y8Fa and (3D)Y8Fa on intestinal transit in mice. (1DME)Y8Fa (0.88 to 22 nmol) inhibited intestinal transit as did F8Famide and morphine. An IP injection of naloxone (2 mg/kg) decreased the morphine effect but had no effect on the response to (1DME)Y8Fa. In contrast, a subthreshold dose of morphine (0.22 nmol) inhibited the response to (1DME)Y8Fa, (3D)Y8Fa delayed intestinal transit only at large dose (22 nmol) but decreased (1DME)Y8Fa and morphine effects at lower ineffective doses. Our findings demonstrate that although F8Famide and morphine could induce the same pharmacological effect, F8Famide receptor activity was modulated by a low-level stimulation of opioid receptors. Furthermore, (3D)Y8Fa should be a useful probe to elucidate neuronal mechanisms controlled by opioids. PMID- 8234021 TI - Comparative study of vascular relaxation and receptor binding by PACAP and VIP. AB - The pharmacological properties of the pituitary adenylate cyclase activating peptides (PACAPs) and vasoactive intestinal peptide (VIP) were compared using: (i) relaxation of vascular and gastric smooth muscle in vitro, and (ii) radioligand binding to membrane preparations of a variety of tissues. Vasoactive intestinal peptide and PACAP-27 were similarly potent in relaxing rat mesenteric arteries, porcine coronary arteries, and rat gastric smooth muscle, whereas PACAP 38 was either more or less potent than the other two peptides depending on the tissue model. Cross-desensitization to relaxation and radioligand binding studies of porcine coronary arteries suggested that VIP and the PACAPs interact with a common receptor in this tissue. A PACAP-preferring receptor with low affinity for VIP was identified in radioligand binding studies of rat brain and anterior pituitary. A second, nonselective, receptor that binds VIP and both PACAPs with high affinity was observed in preparations of rat and porcine arteries and rat lung, liver, brain, and anterior pituitary. PMID- 8234022 TI - Antinociceptive effects of the enkephalinase inhibitor, SCH 34826, in the snail, Cepaea nemoralis. AB - In vertebrates the effects of endogenous opioid peptides are limited by proteolytic enzymes such as endopeptidase 24.11 (enkephalinase), which cleaves the Gly-Phe bonds in both methionine- and leucine-enkephalin. SCH 34826 ((S)-N-[n [1-[(2,2-dimethyl-1,3-dioxolan-4yl) methoxy]carbonyl]-2-phenylethyl]-L phenylalanine-B-alanine) is a potent, highly specific, enkephalinase inhibitor that has marked analgesic effects in mammals. The present study examined the effects of SCH 34826 on opioid-mediated aversive thermal (nociceptive) response of an invertebrate, the land snail, Cepaea nemoralis. SCH 34828 had significant, dose-related antinociceptive effects in Cepaea that were reduced by naloxone and completely blocked by the specific data opiate antagonist, ICI-174,864, and only weakly affected by the specific kappa opiate antagonist nor-binaltrophimine. These findings with SCH 34826 suggest that an enkephalinase similar to that in vertebrates is present and involved in the mediation of opioid (enkephalin) activity in the snail, Cepaea. PMID- 8234023 TI - Role of second messengers in the release of motilin from isolated canine intestinal cells. AB - We examined the intracellular mechanisms for the release of motilin in a preparation of mucosal cells obtained from dog duodenum. Enzymatically dispersed cells were separated by counterflow elutriation to enrich motilin content. Postreceptor activation process was studied by comparing the release of motilin obtained with exogenous analogues or stimulants of the various intracellular signal pathways. Dibutyryl cyclic adenosine monosphate (10(-3) M) and forskolin (10(-5) M) induced a moderate response in motilin secretion. Phorbol ester beta phorphol-12-myristate-13-acetate and phospholipase C were potent stimulants of motilin release. Raising intracellular calcium concentration by calcium ionophore A23187 or increasing calcium content in the incubation milieu failed to modify the secretion of motilin. Analogues of 8-bromoguanosine-3'5' cyclic monosphosphate were ineffective. Therefore, the motilin cell was very sensitive to protein kinase C activators and appeared moderately responsive to a stimulation of the adenylate cyclase pathway. PMID- 8234024 TI - Receptor binding profile of neuropeptide gamma and its fragments: comparison with the nonmammalian peptides carassin and ranakinin at three mammalian tachykinin receptors. AB - The tachykinin binding site preferences of neuropeptide gamma (NP gamma), its C terminal fragments AcNP gamma(3-21), AcNP gamma(5-21), AcNP gamma(7-21), and AcNP gamma(9-21), other mammalian tachykinins, and the nonmammalian tachykinins ranakinin and carassin were examined in membrane binding competition studies. [125I]-Bolton-Hunter [Sar9,Met(O2)11]SP (BHSarSP), [125I]-neurokinin A (INKA) and [125I]-Bolton-Hunter scyliorhinin II (BHScyII) were used to investigate NK-1, NK 2, and NK-3 sites, in rat submandibular gland, gastric fundus, and brain, respectively. Elongation of the neurokinin A molecule does not appear to influence binding to rat tachykinin NK-1 and NK-2 binding sites. Ranakinin has affinity for the NK-1 and NK-2 site similar to that of substance P and neurokinin A, respectively, but has low affinity for the NK-3 site. Despite its structural similarities to neuropeptide gamma, carassin has only moderate affinity for rat tachykinin binding sites. Possession of an acidic residue at position 4 appears critical for binding to rat NK-2 sites. PMID- 8234025 TI - Histones and related preparations interfere with immunoassays for peptide hormones. AB - We report here that histones and certain related preparations generate a consistent interference with radioimmuno (RIA), immunoradiometric (IRMA), and enzyme-linked immunosorbent (ELISA) assays for a number of peptide hormones. Histones H1, H2A, H2B, H3, HIIA, HIIS, protamine, and the related preparations homeostatic thymus hormone and peptide MB35 generated a dose-dependent signal in both the human corticotropin-releasing hormone (CRH) and the human adrenocorticotropic hormone (ACTH) IRMA. This signal was not affected when the linker antiserum was removed from the IRMA reagent mixture, thus proving that the signal was not due to cross-reaction or sample contamination with CRH or ACTH. The above histone preparations, as well as protamine, but not ubiquitin, also generated a strong negative interference with RIAs for ACTH, CRH, rat growth hormone (rGH), and rat prolactin (rPRL). In an ELISA system for the thymic peptide facteur thymique serique, histones and protamine again showed a strong interfering activity. When known amounts of rGH, rPRL, and hACTH were dissolved in charcoal-washed horse serum or supernatants from rat liver homogenates (centrifuged 1 h at 10,000 x g), and the corresponding RIAs and IRMA (for ACTH) were performed in the absence or presence of histones HIIA and HIIS (at 1 mg/ml level), an interfering activity of histones was again observed. We conclude that histones and some related peptide preparations have, when present in biologic fluids, a significant capacity to interfere with peptide immunoassays. PMID- 8234026 TI - The sodium influx stimulating peptide of the pulmonate freshwater snail Lymnaea stagnalis. AB - In Lymnaea stagnalis integumental Na+ uptake is stimulated by the sodium influx stimulating (SIS)-peptide. Its primary structure was determined as: SRTQSRFAS- YELMGTEGTECVTTKTISQICYQCATRHEDSFVQVYQECCKKEMGLREYCEEIYTELPIRSGLWQPN++ +. Antisera raised against parts of SIS-peptide stained neurons in the visceral, parietal, and pleural ganglia, and in the proximal parts of the intestinal, anal, and right internal pallial nerves. Locations and axon projection patterns of these neurons suggest that they represent the previously described neurosecretory yellow cells. PMID- 8234027 TI - Increased neuropeptide Y concentrations in specific hypothalamic nuclei of the rat following treatment with methysergide: evidence that NPY may mediate serotonin's effects on food intake. AB - Neuropeptide Y (NPY) is a potent central appetite stimulant found in hypothalamic neurons that have close anatomical associations with neurons containing serotonin, a powerful anorectic agent. To determine whether the two neurotransmitters interact functionally, we have studied the effects on regional hypothalamic NPY concentrations of acute and chronic administration of methysergide, a 5-HT1BC/serotonin receptor antagonist. Chronic methysergide treatment (10 mg/kg/day) was given by subcutaneously implanted osmotic minipumps (n = 8). Acute effects of methysergide were determined 4 h after a single injection (10 mg/kg) in a separate group (n = 8). Controls (n = 8) had implanted minipumps delivering saline, and also received a saline injection 4 h before sacrifice. Food intake was significantly increased (p < 0.01) by both acute and chronic methysergide treatment. In the chronically treated rats, NPY levels were significantly increased over controls in the arcuate nucleus (ARC; by 41%, p = 0.02) and paraventricular nucleus (PVN; by 40%, p < 0.01). Acute methysergide treatment also increased NPY concentrations in the ARC (by 81%, p < 0.01) and PVN (by 30%, p < 0.01). Methysergide administration, which stimulated feeding, therefore raised NPY concentrations in the ARC, where NPY is synthesized, and in the PVN, a major site of NPY release where NPY injection induces hyperphagia. These findings suggest that NPYergic and serotoninergic innervations in the hypothalamus interact to regulate food intake, and raise the possibility that increased NPY release may mediate the hyperphagic effect of serotoninergic 5 HT1BC/receptor blockade. PMID- 8234028 TI - Pancreastatin decreases plasma epinephrine levels in surgical stress in the rat. AB - Pancreastatin is a novel peptide, isolated from porcine pancreatic extracts, that is known to be derived from chromogranin A. Since chromogranin A-derived peptides have been shown to control secretion from chromaffin cells, we studied the effect of rat pancreastatin, injected intravenously via portal vein, on plasma catecholamine levels in the anesthetized, laparotomized rat. Rat pancreastatin reversibly decreased plasma epinephrine levels, in a dose-dependent manner, without modifying plasma norepinephrine and dopamine levels. These findings suggest that pancreastatin, released from the gastroenteropancreatic system or derived from chromogranin A, may have a role controlling secretion from the adrenal medulla in surgical stress. PMID- 8234029 TI - Activation of CCK-B receptors elevates cytosolic Ca2+ levels in a pituitary cell line. AB - Cytosolic Ca2+ levels ([Ca2+]i) in GH3 cells, a rat anterior pituitary tumor cell line, were monitored with fura-2 by fluorescence measurements. Cholecystokinin octapeptide (CCK-8) produced a transient elevation of [Ca2+]i. The elevation of [Ca2+]i by CCK-8 was inhibited by L-365,260, but not by devazepide. It was still observed when extracellular Ca2+ was eliminated, indicating that CCK-8 mobilizes Ca2+ from intracellular storage sites after interaction with CCK-B receptors. Cholecystokinin octapeptide increased the turnover of phosphatidylinositol, but it did not affect cyclic AMP levels. A possible involvement of phosphatidylinositol breakdown and calcium mobilization in the transduction system of CCK-B receptors in GH3 cells is suggested. PMID- 8234030 TI - Characterization of immunoreactive AVP in the ovine hypothalamo-pituitary axis. AB - The aims of these studies were to determine the precise molecular nature of immunoreactive (IR-) vasopressin (AVP) in the ovine hypothalamo-pituitary axis and to examine a possible role for glucocorticoids in regulating both AVP processing and levels in this axis. The IR-AVP in extracts of paraventricular nucleus, median eminence, portal blood, and anterior and neurointermediate pituitary elutes as a single peak on two distinct HPLC solvent systems, suggesting that AVP is processed identically in these tissues. Identical profiles were also found in extracts from pituitaries and sheep subjected to chronic (10 days) glucocorticoid treatment, or hypothalamo-pituitary disconnection. The latter findings confirm that in sheep, like the rat, AVP is synthesized and processed in the anterior pituitary and is not sequestered from extrapituitary sources. PMID- 8234031 TI - Genetic-based differences in neurotensin levels and receptors in brains of LS x SS mice. AB - Levels of endogenous neurotensin (NT-IR) in the LS x SS RI strains differed by 3.0-, 4.7-, 5.4-, and 6.9-fold in the ventral midbrain (VMB), hypothalamus (HY), nucleus accumbens (NA), and caudate putamen (CP), respectively. Frequency distributions and estimates of the number of genes indicate that differences in NT-IR are polygenically influenced. The NT-IR levels in NA and CP were significantly correlated, but levels in the VMB did not correlate with those in the NA or CP. Specific binding to either low (NTL)- or high (NTH)-affinity receptors as measured in the absence or presence of levocabastine differed significantly in brain regions from among LS X SS mouse strains. Results indicate a polygenic influence mediating the differences in receptor densities and suggest differences in genetic regulation of NTL and NTH receptors. PMID- 8234032 TI - Binding of [3H]angiotensin II and [3H]DuP 753 (Losartan) to rat liver homogenates reveals multiple sites. Relationship to AT1a- and AT1b-type angiotensin receptors and novel nonangiotensin binding sites. AB - The binding characteristics of radiolabeled angiotensin II and the nonpeptidergic angiotensin AT1 receptor antagonist, DuP 753 (Losartan), were studied in rat liver homogenates. Competition experiments with human angiotensin I, II, and III and with the angiotensin antagonists, CGP 42114A, saralasin, DuP 753, and PD123177, confirmed that the [3H]angiotensin II binding was to an AT1-type receptor. Computer analysis of the competition studies using the human angiotensins demonstrated that the data could be best fitted to a model that considers interaction at two sites. Angiotensin II, angiotensin III, and an angiotensin II analogue, [Sar1]angiotensin II, were calculated to have approximately one hundredfold selectivity at each of the two binding sites, but angiotensin I and the antagonists did not show a difference in affinity between the two sites. The addition of 120 mM NaCl and the nonhydrolyzable analogue of GTP, GppNHp (100 microM) to the buffer resulted in a reduction in [3H]angiotensin II binding at both sites. Thus, we suggest that the two sites may represent distinct angiotensin AT1-type receptors, possibly AT1a and AT1b subtypes. The addition of dithiothreitol (DTT) reduced [3H]angiotensin II binding, confirming the binding to AT1-type receptors. Binding studies using the selective AT1 angiotensin II receptor antagonist, [3H]DuP 753, were also performed on the rat liver homogenates. Saturation studies using both angiotensin II and DuP 753 to define nonspecific binding showed that [3H]DuP 753 bound to at least two types of site, one smaller population of receptors that was sensitive to both angiotensin II and DuP 753 and a second site that was sensitive to DuP 753 only.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234033 TI - Effects of thyrotropin-releasing hormone and phorbol ester on dopamine release from dispersed rat tuberoinfundibular dopaminergic neurons. AB - We have investigated the intracellular mechanisms underlying thyrotropin releasing hormone (TRH)-mediated [3H]dopamine ([3H]DA) release from dispersed rat tuberoinfundibular dopaminergic (TIDA) neurons. The specific binding of [3H]Me TRH to these cells is characterized by a single, high-affinity binding site (Kd = 1.2 nM) with a Bmax value of 178 fmol/mg protein. Thyrotropin-releasing hormone markedly increased [3H]DA release and intracellular free calcium concentration ([Ca2+]i) in TIDA neurons, and its effect was abolished by treatment with EGTA (5 mM) or chlordiazepoxide, a specific TRH receptor antagonist (10 microM). Furthermore, to examine the involvement of protein kinase C on [3H]DA release, we investigated the effect of phorbol myristate acetate (PMA), which is known to activate protein kinase C directly. Phorbol myristate acetate induced a significant increase in [3H]DA release in a concentration-dependent manner. Treatment with TRH (1 microM) plus PMA (100 nM) resulted in an additive increase in [3H]DA release. Thyrotropin-releasing hormone (1 microM) still increased [3H]DA release even after preincubation with PMA (500 nM) for 24 h, but PMA (100 nM) did not under the same conditions. These results suggest that TRH may induce DA release in dispersed rat TIDA cells by increasing calcium influx and activating the protein kinase C system. PMID- 8234034 TI - Effects of neonatal blockade of bombesin (BN) receptors with [D-Phe6, phi Leu13 Cpa14]BN(6-14) on adult behavior and sensitivity to BN. AB - Long-term consequences of neonatal blockade of bombesin (BN) receptors were examined in the present study. Rat pups were injected twice daily with [D-Phe6, phi Leu13-Cpa14]BN(6-14), a BN receptor antagonist, at either high (10 mg/kg; HD group) or low (5 mg/kg; LD group) doses from postnatal day 1 through 8. Their behavioral responses to a variety of conditions were compared to those of rats neonatally injected with saline (SAL group) or animals handled but not injected during infancy (UNT group). Adult HD rats entered and spent more time on the open arms of the elevated plus maze than LD, SAL, or UNT animals. Under the conditions of a water deprivation schedule, neither central nor peripheral injections of BN differentiated the neonatally pretreated groups as determined by measures of grooming, feeding, and drinking behaviors. These results indicate that at the dosage regimen employed, neonatal injections of [D-Phe6, phi Leu13-Cpa14]BN(6-14) had little effect on adult sensitivity to BN, but that such treatments could alter activity on the elevated plus maze through as yet unknown mechanisms. PMID- 8234035 TI - Differential N-acetylation of alpha-MSH and beta-endorphin in the intermediate pituitary of the turtle, Pseudemys scripta. AB - Steady-state analyses of the intermediate pituitary of the turtle, Pseudemys scripta, indicated that alpha-MSH-sized immunoreactive forms and beta-endorphin sized immunoreactive forms are major end products of melanotropic cells. Three forms of alpha-MSH-related immunoreactivity were detected. The two major forms had the same reversed-phase HPLC properties as synthetic N,O-diacetyl-ACTH(1-13) NH2 and N-acetyl-ACTH(1-13)-NH2. These forms accounted for 97% of the total alpha MSH-related immunoreactivity detected. A minor peak of ACTH(1-13)-NH2 was also detected. Multiple forms of beta-endorphin-related immunoreactivity were detected, which varied in net positive charge (+1 to +5), apparent molecular weight (2.4 to 3.5 kDa), and degree of N-terminal acetylation. Although N acetylated forms of beta-endorphin were detected in the turtle intermediate pituitary, the major forms of turtle beta-endorphin were nonacetylated. These features of the turtle intermediate pituitary POMC-specific N-acetylation mechanism are similar to, yet distinct from, the POMC N-acetylation mechanisms observed for mammals. These data suggest that POMC-specific N-acetylation mechanisms were present in reptiles prior to the divergence of the anapsid and synapsid lines. PMID- 8234036 TI - POMC-derived peptide immunoreactivity in neural lobe axons of the human pituitary. AB - The efferent projections of proopiomelanocortin (POMC) neurons in the arcuate nucleus and nucleus of the solitary tract have been extensively characterized in the rat, but are less well understood in the human brain. We report here that ACTH, alpha-MSH, beta-endorphin, and N-acetyl-beta-endorphin immunoreactive axons are localized in the neural lobe of the human pituitary gland, in congruence with prior evidence that beta-endorphin and other POMC-derived peptides modulate vasopressin and oxytocin secretion. PMID- 8234037 TI - [Progress in diagnosis of viral hepatitis]. PMID- 8234038 TI - [Markers of hepatitis A infection in drug addicts]. AB - Prevalence of HAV infection markers was studied in 100 drug addicts whose sera were collected between 1988 and 1989. Anti-HAV antibodies were found in 65 (65%) of the tested drug addicts and in 55% of the individuals serving as the control group. No correlation between the presence of HAV infection markers, and sex, duration of drug abuse or HIV status was seen. However, drug addicts with anti HAV antibodies were older than those without these antibodies. There was no difference in a mean titre of anti-CMV and anti-HSV type 1 antibodies between the individuals with and without HAV infection markers. It suggests that the tested markers are specific for HAV infection. PMID- 8234039 TI - [Evoked potential test in patients with acute and chronic liver diseases]. PMID- 8234040 TI - [Activity of angiotensin converting enzyme I and levels of acid alpha glycoprotein in selected liver and biliary tract diseases]. AB - Differential value of ACE activity and acid alpha 1-glycoprotein was evaluated in the selected liver and biliary tract diseases. The study involved 75 patients divided into 4 subgroups, according to the character of their disease: patients with the acute viral hepatitis, chronic viral hepatitis, liver cirrhosis, and cholelithiasis. It was found that ACE activity was significantly increased in all pathologies involving liver parenchyma, and normal in patients with extrahepatic cholestasis. It was also shown that simultaneous assays of ACE and acid < alpha 1 glycoprotein may serve as a sensitive test differentiating jaundice in parenchymal hepatic diseases from that in the course of extrahepatic pathology. PMID- 8234041 TI - [Evaluation of microsomal liver enzyme function in patients with duodenal ulcer treated with ranitidine]. AB - Functioning of hepatocytes has been assessed with respiratory test with aminopyrine labelled with 14C radioisotope in patients with duodenal ulcer treated with ranitidine. Reversibility of changes in liver microsomal system at different intervals after the completion of therapy has also been evaluated. Altogether 30 patients with duodenal ulcer treated with ranitidine have been examined: prior to the treatment--group Ia, after one day of ranitidine administration--group Ib, within 4 weeks of therapy--group Ic, within 8 weeks of therapy--group Id, and one week after the completion of the treatment--group Ie, as well as after a 4-week follow-up period--group If. Significant decrease in the results of respiratory test has been noted in patients treated with ranitidine after both four and eight weeks. Activity of liver mono-oxidases approached baseline values after one week following the completion of therapy. The results of routine liver functioning tests in all patients of groups I and II have been within normal values. The obtained results suggest that patients treated with ranitidine should cautiously be given other medicines, especially those affecting liver functioning or metabolized in the liver. PMID- 8234042 TI - [Liver damage in drug addicts]. PMID- 8234043 TI - [Changes of certain biochemical parameters of liver function in patients after allogenic bone marrow transplantation]. AB - The disturbances in bilirubin level, transaminases (GOT, GPT) and alkaline phosphatase (AP) activities in serum of patients undergoing allogeneic bone marrow transplantation are discussed. Hyperbilirubinemia was considered in all, increase in transaminases activity in half of the group of 10 patients after marrow infusion. In the patients displaying hemopoietic reconstitution bilirubin levels returned to normal. In case of bone marrow graft failures or severe complications in posttransplant period the profound abnormalities in tested parameters were encountered, particularly before death. Possible causes of such disturbances in biochemical hepatic parameters are also briefly discussed. PMID- 8234044 TI - [Evaluation of interferon alpha in treatment of chronic viral hepatitis B]. PMID- 8234045 TI - [Endogenic hepatic coma in the course of acute hepatitis treated with arterialization of portal blood]. AB - Symptoms of endogenic hepatic coma were observed in the course of acute hepatitis in 17 patients admitted to the I Clinic of Infectious Diseases of Silesian Medical School between 1987 and 1992. Five of them were treated with the arterialization of portal blood. At least one exchange transfusion preceded the arterialization in four cases. Recovery was obtained in 3 patients. Two patients died because of complications which occurred during the twenty-four hours after the intervention. In the first case the reason of the death was the extensive myocardial infarction, in the second one-DIC and ARDS. As it has been observed, the prothrombin rate should not be lower than 30% in these patients who are to undergo the arterialization of portal blood. This value of the prothrombin rate is provided by at least one exchange transfusion. PMID- 8234046 TI - [Hydrothorax as a major symptom of decompensated liver cirrhosis]. AB - A case of a 54-year patient with decompensated liver cirrhosis is presented. As a major symptom rapidly increasing hydrothorax on the right side without marked ascites was observed. Diagnosing and simultaneously establishing etiology of hydrothorax on the base of clinical and laboratory findings was a difficult process. Imaging tests did not show any signs of splenomegaly. PMID- 8234047 TI - [Current classification of viral hepatitis with particular reference to non-A, non-B type]. PMID- 8234048 TI - [Hemodynamic problems in portal hypertension]. PMID- 8234049 TI - [Mechanism of cell infection with HIV]. AB - The mechanisms of HIV infection of target cells are described. Particularly the role of three types of cell receptors, which participate in the process of virus cell interactions: CD4 protein, receptor for Fc fragment of antibodies and complement receptors are discussed. The variability of the virus variants, which determines the virus tropism and cytotoxic properties towards specific cell types is stressed. PMID- 8234050 TI - [Appearance of thrombocytopenia in patients infected with HIV]. AB - Results of quantitative determinations of thrombocytes and selected, basal immune parameters in 263 HIV infected subjects are presented. A decrease in the platelet count was observed more frequently in symptomatic subjects, including AIDS patients, than in asymptomatic HIV carriers or patients with generalized lymphadenopathy only. A significant correlation between thrombocyte number and percentage and number of CD4+ lymphocyte and CD4+/CD8+ lymphocyte ratio was found only in the group of symptomatic subjects. The mechanisms responsible for thrombocytopenia in HIV infected patients ore discussed in details. PMID- 8234051 TI - [Labial herpes in patients with affective disorders receiving long-term lithium carbonate]. AB - Lithium carbonate has been administered to 69 patients (45 women and 24 men) for 1-17 years as affective disorders prevention. Its effect on the recurrence and clinical course of labial herpes infection has been analysed both prior to and after the administration of lithium carbonate. Labial herpes has been diagnosed in 28 patients before lithium prophylaxis. The drug significantly decreased virus infection recurrence incidence in this group. No labial herpes recurrence has been noted in 13 patients after the treatment. Lithium efficiency has not been dependent on patients' age, duration of therapy, and lithium levels in both blood serum and erythrocytes. These results suggest, that lithium salts may be effective in certain herpes simplex infections at doses used for prevention affective disorders. PMID- 8234052 TI - [Complications of monocytic angina in children]. AB - The studies aimed at assessing the kind and the incidence of the complications of monocytic angina in childhood. In 7 out of 343 hospitalized children neurological disorders were found, thrombocytopenia was seen in 5 cases, pneumonia in 3 cases, bronchitis in 12 cases, and AV block in 3 cases. The most frequent complication of monocytic angina was respiratory tract obturation (6.4% of cases). PMID- 8234053 TI - [Incidence of measles in vaccinated children]. PMID- 8234054 TI - [Evaluation of preventive vaccination against rabies performed at the Department of Parasitic and Occupational Diseases in Bialystok in 1989-1990]. AB - Between 1989-1990, 502 individuals exposed to the animals with rabies or suspected of rabies reported to out-patient clinic of agricultural occupational diseases at the K. Dluski Hospital and to the Department of Parasitic and Occupational Diseases in the Medical Academy in Bialystok. Sixty one (12.2%) of these individuals were vaccinated. Most frequently these people were attacked by dogs (70.4%), cats (11.4%), rats (6.5%), and foxes (3.2%). Rabies was diagnosed only in 9.8% of cases (Negri bodies in the brain and positive immunofluorescence test.). A vaccine inactivated on the human diploid tissue (Institute Merieux) was applied without any postvaccination complications. Transient micro-macular rash on face and chest of 3 patients, and local inflammatory reaction in 2 patients did not disturb vaccination process. Active-passive immunization was used in case of extensive lesions. PMID- 8234055 TI - [CMI multitest in assessment of cell-mediated immunity in patients with chronic tonsillitis]. AB - Cell-mediated immunity has been assessed in 20 patients with chronic tonsillitis with CNI multitest. The test was performed both prior to and after tonsillectomy. A reduced skin reactivity, especially after surgery, has been observed in patients as compared with a control group. PMID- 8234056 TI - [Ultrasound examination in presentation A for diagnosis of maxillary sinusitis in childhood]. AB - Maxillary sinusitis is frequent in childhood. This study aimed at evaluating the value of ultra sound in presentation A in the diagnosis of maxillary sinusitis in childhood. Altogether 324 maxillary sinuses in 162 children were examined. A value of ultra sound examination was verified comparing the obtained results with the results of X-rays, considering three classifications of the latter. Besides the attempt of correlation of the results, ultra sound specificity and sensitivity were evaluated. Relatively high sensitivity and relatively low specificity of ultra sound technique were noted. It seems that ultra sound examination in presentation A may serve as a screening technique in children. Being sensitive and safe, this technique may be used in patients who require frequent examinations in persisting chronic disorders or recurrent maxillary sinusitis. However, the obtained results should be verified with X-rays because of the low specificity of ultra sound examination, especially prior to therapy of patients with unclear clinical picture of the disease. PMID- 8234057 TI - [Phage and serological types of bacteria infecting patients in anesthesiology and intensive therapy (1988-1990)]. AB - 1736 of biological materials, being taken from 264 patients, were investigated since 1988 to 1990. 1410 kinds of microorganisms were cultured from 999 biological materials, in which the growth of bacterial flora was noticed. Following species were isolated most frequently: Pseudomonas aeruginosa 15.39%, Proteus mirabilis 12.91%, Klebsiella pneumoniae 10.43% and Staphylococcus aureus 10.43%. The most frequent serological type according to Fisher's scheme was Pseudomonas aeruginosa--immunotype T 3.7 and according to Habs scheme--immunotype P 16. Strains of Staphylococcus aureus were most frequently sensitive to phages the group II. In case of Klebsiella sp. bacilli, the most predominant strains were not typed either by basic or extended phage sets. PMID- 8234058 TI - [Cefuroxime axetil--efficiency and safety for treatment of upper airways and middle ear infections in children]. AB - The study aimed at assessing the clinical efficiency, safety, and tolerance of cefuroxime axetil suspension in the treatment of children with the acute upper respiratory infections and/or the acute otitis media. The trial was open, multicenter, involving 304 children aged between 3 months and 12 years. They were recruited from 18 general practice centers in Poland. Children were given cefuroxime axetil suspension in the dose of 10 mg/kg body weight (upper respiratory) or 15 mg/kg otitis media. max. 250 mg) bid. Children were examined prior to the treatment, 3-4 days following the start of therapy, 1-2 days after completion of the treatment, and followed-up for 14 days. Post-therapy examination has shown 93% cure rate. During the follow-up period 0.77% of patients relapsed. Only minor adverse reactions were reported by 4.9% of patients. Most common complaint was vomiting. Cefuroxime axetil suspension was safe and effective therapy in the acute upper respiratory infections and the acute otitis media in childhood. PMID- 8234059 TI - [Hydatid cyst]. PMID- 8234060 TI - [Attacks on people by Argas (A) reflexus (Ixodida, Argasidae)--harmful parasites for humans and animals]. PMID- 8234061 TI - [Infection with parvovirus B 19 virus in humans]. AB - Human parvovirus B 19 (B 19) causes endemic infections in Germany. Most frequently. B 19 infection is associated in with erythema infectiosum. In patients with underlying chronic-haemolytic anaemia, B 19 infection can result in aplastic crisis. In patients with hereditary or acquired immuno-deficiency syndromes, B 19 virus may persist causing chronic bone marrow aplasia. In pregnancy B 19 infection may cause hydrops fetalis resulting in fetal loss. B 19 infection may cause hydrops fetalis resulting in fetal loss. B 19 infection in sometimes associated with vascular purpura. Infections occur mostly as epidemic outbreaks in families, kindergartens and schools as well as hospital wards. Acute or past B 19 infections are confirmed by detection B 10-specific antibodies (IgM/IgM) by ELISA. PMID- 8234062 TI - [Tetanus--remarks after 50 years of clinical experience]. PMID- 8234063 TI - [HIV infection and acquired immunodeficiency syndrome (AIDS) in France--warning for Poland]. PMID- 8234064 TI - [What does periodontology concern itself with?--overview of problems]. PMID- 8234065 TI - LY207320 (6-methylene-4-pregnene-3,20-dione) inhibits testosterone biosynthesis, androgen uptake, 5 alpha-reductase, and produces prostatic regression in male rats. AB - LY207320 is an in vitro inhibitor (estimated IC50 = 0.06 microM) of steroid 5 alpha-reductase that catalyzes the conversion of testosterone (T) to dihydrotestosterone (DHT). In contrast, LY207320 was only moderately active against rat prostatic 5 alpha-reductase in vivo (32% inhibition at 50.0 mg/kg single dose). LY207320 did, however, inhibit the in vivo uptake of [3H]-T by the prostate. The antiprostatic and endocrine effects of this agent were evaluated following daily (21 days) administration to castrated, androgen-supplemented castrate, and intact rats. LY207320, which has modest progestational competitive binding activity, does not bind to rat prostatic androgen or uterine estrogen cytosolic receptors. In the castrated male rat, subcutaneously (s.c.) administered LY207320 had no androgen agonist activity, as evidenced by a lack of accessory sex organ weight gains. Administration of s.c. LY207320 to intact rats for 21 days at doses greater than 5.0 mg/kg-day produced significant (P < 0.05) reductions of seminal vesicle and ventral prostatic weights (maximal regression = -65% and -40% from control values, respectively at 50.0 mg/kg-day). The compound had no regressive activity on male accessory sex organs when administered orally. LY207320 did not alter circulating prolactin, LH, or corticosterone levels, but at high doses (> or = 50.0 mg/kg-day), lowered circulating T[-67% from intact control levels (P < 0.05)]. Histological analysis of the rat ventral prostates (RVPs) in LY207320-treated rats was consistent with an androgen-deprived state. Decreased circulating androgens and prostatic regression are associated with inhibition of testicular 17 alpha-hydroxy/C17,20-lyase enzyme activity (IC50 = 0.06 microM). These findings support the contention that LY207320 is a physiological antagonist of androgen action in male rats, and that its effects are mediated primarily through inhibition of testicular androgen production rather than accessory sex organ 5 alpha-reductase. PMID- 8234066 TI - Presence of 11 beta-hydroxysteroid dehydrogenase enzyme in the human prostate tumor cell line LNCaP. AB - 11 beta-Hydroxysteroid dehydrogenase (11 beta-HSD) is the enzyme that catalyzes the reversible oxidation of the biologically active steroid cortisol and corticosterone to their inactive metabolites cortisone and dehydrocorticosterone. We report its presence in significant levels in the human prostate carcinoma cell line LNCaP cultured in medium RPMI-1640 with 10% fetal calf serum (FCS). The 11 dehydrogenase activity of 11 beta-HSD is present, while the 11-reductase activity is undetectable in these tumor cells under the present culture conditions. The enzyme activity is found to be linear with time of incubation, and is proportional to plated cell density. The enzymatic activity can be determined in cultures maintained for longer times. Carbenoxolone, the potential inhibitor of the 11 beta-HSD, inhibits 95% of the dehydrogenase activity of the tumor cells when used in nM concentration. The presence of this enzyme in tumor cell line indicates that 11 beta-HSD plays an important role in maintaining the active glucocorticoid levels in the prostate. PMID- 8234067 TI - Endocrine and antiprostatic effects of raloxifene (LY156758) in the male rat. AB - The benzothiophene anti-estrogen, raloxifene [LY156758; (6-hydroxy-2-(4 hydroxyphenyl) benzo(b)thien-3-yl)(4-(2-1-piperidinyl)ethoxy)phenyl methanone hydrochloride] has selective estrogen pharmacological antagonist activity in female rats. The present studies were done in the male rat to assess activity of raloxifene related to inhibition of prostatic growth and effects on the hypothalamic-pituitary-gonadal axis. Raloxifene did not compete for binding of the androgen, [3H]-methyltrienolone (R1881) in cytosolic extracts of ventral prostate. Similarly, the compound did not inhibit prostatic 5 alpha-reductase or testicular 17 alpha-hydroxy/C17,20-lyase activities. Raloxifene had no effect on the ventral prostatic uptake of [3H]-R1881 in vivo. Administration of estradiol to castrated male rats stimulated fourfold increases of in vitro ventral prostatic binding of [3H]-R1881. Raloxifene was devoid of agonist activity in castrated animals, because the compound had no stimulatory effect on prostatic androgen receptor binding activity. When raloxifene was coadministered with estradiol, the compound markedly antagonized the estrogen-induced increase of prostatic [3H]-R1881 binding, confirming its antiestrogenic properties in male rats. Serum prolactin was also elevated significantly (P < 0.05) with a single injection of raloxifene (20.0 mg/kg). In these same animals, serum FSH was significantly (P < 0.05) decreased by one dose (10.0 mg/kg) of the compound. Luteinizing hormone levels in castrated male rats were unaffected by raloxifene administration. Raloxifene treatment of castrated males significantly (P < 0.05) antagonized the stimulatory response of the ventral prostate (VP) to exogenous androgens in a dose-dependent manner. Raloxifene treatment of intact male rats for 14 and 28 days produced significant (P < 0.05) dose-dependent regression of the VP and seminal vesicles (SV). The VP regressive responses to raloxifene were associated with a decline in serum testosterone levels. Histological analysis of the VPs in raloxifene-treated rats was consistent with an androgen-deprived state. These findings support the contention that raloxifene is a pure estrogen antagonist and a physiological antagonist of androgen action in male rats. These pharmacological properties provide support for further structure-activity and mechanistic investigations with benzothiophenes in the medical management of prostatic neoplasia. PMID- 8234068 TI - Heat shock protein response in a prostate tumor model to interstitial thermotherapy: implications for clinical treatment. AB - Hyperthermia is being utilized individually and in conjunction with other therapies in treating malignant and benign tumors, though few studies have examined cellular effects of elevated temperatures in the prostate model. Highly conserved proteins of the 70 kDa heat shock protein family (HSP 70) are produced in response to environmental stresses, including heat, and are found in all organisms. HSPs are an indicator of cell damage, are associated with thermotolerance, and provide cells with transient resistance to subsequent thermal challenges. Transient thermotolerance is important in the determination of temperature, duration, and sequencing for treatments. This preliminary study analyzes the HSP 70 response of the Dunning R3327 adenocarcinoma model to a single 50 degrees C 1 hr treatment. Elevated HSP levels were found between 10 and 16 hr, returning to baseline by 24 hr. As some fractions of the cells are able to produce HSP 70 following treatment, the data suggest that currently utilized clinical temperatures (42-46 degrees C) administered for 1 hr are inadequate. HSP levels in response to hyperthermia, radiation, and chemotherapy may be useful in finding optimal treatment regimens for prostate cancer. PMID- 8234069 TI - Arthroscopic management of rotator cuff disease. AB - The etiology of rotator cuff disease is controversial, but is not always secondary to cuff impingement between the humeral head and the coracoacromial arch. Rotator cuff disease, and not impingement syndrome, more accurately describes this process. The spectrum of rotator cuff disease begins with rotator cuff inflammation (Stage I) and progresses to tendinitis (Stage II) and partial or full thickness tears (Stage III). Refractory Stage II and III disease without rotator cuff tears undergoes subacromial bursectomy and acromioplasty. Partial thickness tears are debrided with an acromioplasty, bursectomy, and removal of inferior clavicular osteophytes. Full thickness tears are treated arthroscopically or open, depending on the tear size and the patient expectations. In general, tears from 0 to 5 cm in active patients undergo subacromial decompression and a mini-open repair. Low demand patients with 0 to 5 cm tears or tears greater than 5 cm are treated with subacromial decompression and rotator cuff debridement. PMID- 8234070 TI - Arthroscopically assisted repair of acute Bankart lesion. AB - Acute traumatic anterior shoulder dislocations are associated with high recurrence rates in young athletic individuals. Arthroscopically assisted stabilization of acute initial traumatic anterior dislocation was carried out in 11 young athletes. The average age was 20 years, with nine males and two females. The anterior acute Bankart lesion was surgically repaired using the technique described by Caspari/Morgan. There were no surgical complications. All patients were able to return to their sport without restriction. One patient had a transient episode of instability 8 months postoperatively associated with trauma, but no further sequelae. Using the Rowe scale there were nine excellent and two good results. Using repair of a Bankart lesion provides a stable shoulder with minimal loss of function and low recurrence rate. Early intervention should be considered in young, highly competitive athletes. PMID- 8234071 TI - Current concepts in ankle arthroscopy. AB - Ankle arthroscopy has rapidly become an important diagnostic and therapeutic procedure. Currently, indications for operative arthroscopy include transchondral talar dome fractures, acute articular fractures with hemarthrosis, posttraumatic synovitis, loose bodies, inflammatory synovitis, degenerative joint disease, and soft tissue impingement. Diagnostic arthroscopy is indicated for the patient with a chronically painful, symptomatic ankle when nonoperative treatment has failed and other measures have failed to produce a diagnosis. Three standard portals are used for routine ankle arthroscopy and allow a systematic examination of the joint. Mechanical distraction may be required to visualize the entire joint, the tight ankle, the ankle with posterior lesions, or to allow operative instruments to be introduced. The use of lasers in arthroscopy has yet to be clearly defined. The small size of the laser is an advantage in the ankle, but cost remains a disadvantage. Advances in technique and equipment will continue to expand the indications for this procedure. PMID- 8234072 TI - Advances in elbow arthroscopy. AB - Arthroscopy of the elbow is a very precise and demanding technique. Because of the proximity of neurovascular structures to the recommended portals, a thorough knowledge of regional and intraarticular anatomy is essential. The high degree of congruency of the articular surfaces and the relatively small capsular volume make sound arthroscopic skills a necessity. With proper precautions, the technique is safe and affords the opportunity to obtain valuable diagnostic information and to perform selected procedures without the morbidity of an arthrotomy. PMID- 8234073 TI - Arthroscopic categorization of intercarpal ligamentous injuries of the wrist. AB - Recent advances in our knowledge of wrist ligamentous anatomy, as well as improvements in arthroscopic techniques, permit the diagnosis and treatment of a wide variety of wrist injuries. The authors review current concepts in wrist ligamentous anatomy, patterns of instability, and the role of the arthroscope in the evaluation and treatment of such instability. Specifically, wrist portal anatomy and techniques of arthroscopic treatment of patients with partial and complete intercarpal ligament injuries are described. Chronic complete injuries are a contraindication of arthroscopic treatment. The authors conclude that wrist arthroscopy provides a valuable method of diagnosing and treating partial and complete intercarpal ligamentous injuries. PMID- 8234074 TI - Arthroscopic management of wrist triangular fibrocartilage complex injuries in the athlete. AB - Wrist arthroscopy can facilitate early definitive diagnosis of debilitating soft tissue injuries in athletes. Many such injuries can be treated successfully with minimally invasive arthroscopic techniques, reducing the morbidity associated with surgical exposure and permitting earlier return to competition. The triangular fibrocartilage complex (TFCC) is vulnerable to injury from rotational forces or axial load applied to the hand. Under arthroscopic control, injuries to the central articular disk can be treated by excision of unstable tissue fragments; peripheral separation of the disk from the dorsal or volar ligaments can be repaired with sutures to achieve complete healing. Avulsion fractures from the dorsal ulnar margin of the sigmoid notch of the radius are better treated through a small arthrotomy after initial arthroscopic evaluation. PMID- 8234075 TI - Orthopedic rehabilitation: the legacy of Dr Nickel. PMID- 8234076 TI - Radiologic case study. Macrodystrophia lipomatosa. AB - Macrodystrophia lipomatosa is a rare form of gigantism involving an extremity consisting of predominately fibroadipose tissue. Radiographically, it appears as hypertrophy of soft tissues and bone. CT and MR scanning delineate the tissues involved and are virtually diagnostic, demonstrating the predominantly adipose component as well as the overgrowth of the bones, nerves, and vessels also involved. PMID- 8234077 TI - Endoscopic ACL reconstruction. AB - The endoscopic technique offers the advantage of one incision and a femoral osseous tunnel trajectory that is more in line with the collagen fibers of the graft. Technically, it is more demanding to reproducibly obtain the interference fixation than using the two-incision technique. There is a tendency for screw and graft divergence in the femoral tunnel. The exact clinical significance of this screw and bone plug divergence has yet to be clarified. Surgeons must assess their ability to obtain the best results for the patient. There is a definite learning curve for the endoscopic technique. We believe that, with further advances in instrumentation, fixation, and alternative grafts, it will eventually be the approach of preference for ACL reconstructions. PMID- 8234078 TI - Technique of arthroscopically assisted PCL reconstruction. PMID- 8234079 TI - Current concepts in meniscal repair. AB - As the importance of meniscal preservation has become recognized, meniscal repair has become a more commonly practiced procedure. We briefly review the current, commonly accepted indications, techniques, and our rehabilitation protocol for meniscal repair. Both open and arthroscopic repairs are reviewed, including the arthroscopic techniques classified as inside-out, outside-in, and all inside. We conclude with a review of the results of meniscal repair and some thoughts on future directions. PMID- 8234080 TI - Complications in arthroscopic surgery of the knee and shoulder. AB - Arthroscopy is a valuable technique used by the majority of orthopedic surgeons in practice. Complications in arthroscopy have been compiled on a formal basis since 1983. Several studies on complications are reviewed and summarized. A recent prospective study on complications is reviewed in detail. The most frequent type of complication encountered in arthroscopic surgery is hemarthrosis. The procedure with the highest complication rate is the lateral retinacular release. Technical considerations are discussed which would be useful in lessening the incidence of complications in arthroscopy. PMID- 8234081 TI - Arthroscopic management of instability of the shoulder. AB - Anterior shoulder dislocation in the young athlete may be difficult to treat and, without proper care, usually results in recurrent episodes of instability. By permitting direct visualization of all intraarticular pathology, the arthroscope assists in the diagnosis and helps to determine appropriate therapeutic interventions. Numerous techniques have been developed for arthroscopic reconstruction of unstable shoulders. These can be classified according to the type of fixation employed. The categories include metal devices such as staples or screws, absorbable devices, and suture-based methods, both anterior and transglenoid. An anterior suture anchor method using a new suture passing device known as a Shuttle Relay (Linvatec Corp, Largo, Fla), which allows the surgeon to incorporate non-absorbable mattress sutures in the repair, is described in detail. PMID- 8234082 TI - Population reports. Controlling sexually transmitted diseases. AB - While AIDS seizes the headlines, other sexually transmitted diseases (STDs) create devastation of their own. In women STDs can lead to pelvic inflammatory disease, causing lifelong pain, infertility, and ectopic pregnancy, which can kill. Children are born with blinding eye infections. Men are left infertile. People die of advanced stages of syphilis. Furthermore, STDs multiply the transmissibility of HIV, the AIDS virus, as much as ninefold. PMID- 8234083 TI - [Chromosomal aberrations in human sperm]. PMID- 8234084 TI - [Mechanisms of cellular cytotoxicity. I. Stages of the lytic process]. PMID- 8234085 TI - [Mitotic cyclins--new possibilities for examining mechanisms of neoplasm growth]. AB - Mitotic cyclins constitute a regulatory subunit of the histone H1 kinase complex. On the ground of their primary structure they are divided into two classes A and B, both necessary for the mitosis. Cyclin A activates histone H1 kinase and becomes destroyed by proteolysis earlier than cyclines B and plays an important role in the DNA replication. Cyclins A and B may be involved in the development of neoplastic disorders either directly (inappropriate expression of the cyclin A gene caused by hepatitis B virus in hepatocellular carcinoma, or interactions of this cyclin with factors participating in the regulation of cell proliferation) or indirectly by phosphorylation of some oncogene or antioncogene proteins by a cdk (cyclin dependent kinase). PMID- 8234086 TI - [Role of melatonin in human physiology and pathology. I. Circadian rhythm of melatonin secretion. Involvement of melatonin in reproductive physiology. Melatonin and hypothalamic-pituitary-adrenal cortex axis]. AB - This paper summarizes the recent knowledge on the role of melatonin in humans. The first part of the present review considers the following aspects: circadian rhythm of melatonin excretion, melatonin in body fluids, melatonin influence on sexual maturation, changes of melatonin blood concentration during the menstrual cycle and pregnancy, and the influence of melatonin administration on prolactin concentration. Current views on the relationship between melatonin and the hypothalamo-pituitary-adrenal cortex axis, as well as on changes of melatonin concentration during hypothyreosis, hyperthyreosis, and hyperparathyroidism are also discussed. PMID- 8234087 TI - [Yeast model of an eucaryotic organism in molecular biology]. PMID- 8234088 TI - [Sequencing the genome of yeast Saccharomyces cerevisiae. Aims, strategies and results]. PMID- 8234089 TI - [Function and biogenesis of peroxisomes in yeast Saccharomyces cerevisiae]. PMID- 8234090 TI - [Biosynthesis of heme in yeast Saccharomyces cerevisiae]. PMID- 8234091 TI - [Use of yeast mutants for examining common biosynthetic pathways of dolichols, sterols and ubiquinones]. PMID- 8234092 TI - [Genetic regulation of N-glycosylation in yeast protein]. PMID- 8234093 TI - Toxic shock syndrome. Are you recognizing its changing presentations? AB - Nonmenstrual cases of toxic shock syndrome now occur at least as often as the classic menstrual cases. The syndrome has been reported in every conceivable clinical setting involving colonization or infection with Staphylococcus aureus and as a complication of surgical procedures, traumatic injuries, and local infections. Unusual clinical manifestations have been observed in patients with AIDS. Cases of toxic shock syndrome also have been linked to infection with toxin producing strains of coagulase-negative staphylococci and group A streptococci. Knowledge of the expanding spectrum of toxic shock syndrome and early consideration of the disease in patients with fever, hypotension, rash, and/or multisystem disease are essential for timely diagnosis and optimum management. PMID- 8234094 TI - Rhabdomyolysis after exercise. AB - Rhabdomyolysis should be considered in the differential diagnosis of acute muscle weakness or pain in patients who have recently increased their level of exercise, no matter what their previous level of physical fitness. Measurement of an elevated creatine kinase level confirms the diagnosis. Management of rhabdomyolysis is aimed at avoiding acute renal failure through aggressive fluid replacement to maintain a high urine output. PMID- 8234095 TI - The 'Salesman-in-chief' sells a health plan. PMID- 8234096 TI - Does rheumatoid factor always mean arthritis? AB - Rheumatoid arthritis is a clinical syndrome, and the diagnosis requires the presence of pain, swelling, and tenderness in the joints. In the absence of these features, identification of rheumatoid factor in the serum is of little use. Because rheumatoid factor is an immune complex, it is a marker of immune activation and, therefore, may be present in the circulation of persons with a variety of inflammatory conditions that stimulate the immune system. Also, 5% of the healthy population have significant titers of rheumatoid factor in their serum. PMID- 8234097 TI - Germ warfare. PMID- 8234098 TI - Gallstones. Who are good candidates for nonsurgical treatment? AB - Although surgery remains the ultimate treatment for symptomatic gallstone disease, the number of nonsurgical alternatives continues to grow. It is important to consider these options. The patient's age and overall health and the composition, number, and size of gallstones are major factors in determining whether alternative methods of therapy are appropriate. Many patients are eliminated from consideration when criteria for nonsurgical therapy are carefully followed. For example, oral dissolution therapy is relatively effective, but only about 20% to 25% of patients are candidates. PMID- 8234099 TI - Biliary obstruction. Nonsurgical treatment with endoscopic and radiologic techniques. AB - Several methods and combinations of techniques are available to treat biliary obstruction. The approach to treatment should be determined by the patient's situation and the expertise available in the various treatment options rather than by adherence to a standard plan. Clinical outcomes reported in the literature reflect results achieved by experts, and traditional surgical approaches may be most appropriate if experience with endoscopic and radiologic techniques is lacking in the patient's community. Various methods can often be used in a complementary or sequential fashion to provide optimal patient care. PMID- 8234100 TI - Primary sclerosing cholangitis and primary biliary cirrhosis. How effective is medical therapy? AB - Although a great deal of investigation has led to some novel approaches in treating primary sclerosing cholangitis (PSC) and primary biliary cirrhosis (PBC), no method has been proven to definitively alter the long-term outcome of either disease. In addition, many of the medical regimens reviewed in this article have serious toxicity, which limits their widespread use. Furthermore, the natural history of PSC and PBC may vary widely between individual patients, complicating the decision of whom and when to treat. Ursodeoxycholic acid (Actigall) appears to be a safe and possibly effective agent for both diseases in their early stages, and its use in selected patients may be warranted. Methotrexate (Folex, Mexate, Rheumatrex Dose Pack) may improve liver enzyme levels in some patients, but it is hepatotoxic and should be used cautiously. The general application of toxic immunosuppressive agents should await the results of large, long-term, prospective trials. Liver transplantation is the only proven effective therapy for advanced, decompensated PSC and PBC. PMID- 8234101 TI - Brain stethoscopes: the use and abuse of brief mental status schedules. PMID- 8234102 TI - Current concepts of neuropsychiatric systemic lupus erythematosus (NP-SLE). PMID- 8234103 TI - Serum lipids and coronary heart disease in British elderly. AB - To assess the relationship between serum lipids and coronary heart disease (CHD) in an elderly population (mean age = 77.4; SEM +/- 0.4; range 70-92 years), serum total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) were measured in 84 (males = 52) subjects with evidence of definite myocardial infarction and 125 (males = 77) age- and sex matched controls free of CHD on Rose questionnaire and resting electrocardiogram. On multivariate logistic regression analysis, CHD showed inverse association with HDL-C (odds ratio for decrease by 1 s.d. = 1.6, P = 0.02) in males and positive association with TC (odds ratio for increase by 1 s.d. = 1.8, P = 0.03) and LDL-C (odds ratio for increase by 1 s.d. = 1.8, P = 0.04) in females. Of the other variables only diabetes in males (odds ratio 2.4, P = 0.05) and left ventricular hypertrophy in females (odds ratio 8.8, P = 0.03) were associated with CHD. The results suggest that further prospective studies to further evaluate the relation of serum lipids with CHD in this age group are indicated. PMID- 8234104 TI - Right-sided endocarditis in the non-drug addict. AB - Fifteen patients with right-sided infective endocarditis during a 5 year period (1985-1990) were retrospectively reviewed. Isolated tricuspid valve involvement occurred in nine patients. Staphylococcus aureus was the causative organism in seven cases; four were culture negative. The diagnosis was established by two dimensional echocardiography in 11 patients and at postmortem in the remaining four patients who succumbed shortly after admission. Fever, tachypnoea and pneumonia were universal features. A successful outcome ensued in eight patients with medical therapy alone and in two patients who were submitted to valve replacement. Five patients died, two from uncontrolled infection with repeated pulmonary emboli. Right-sided infective endocarditis should be suspected in any pneumonic illness that complicates post-abortal infection or other inadequately treated sepsis. Two-dimensional echocardiography is important in diagnosis since cardiac signs are minimal at presentation. PMID- 8234105 TI - Atypical presentations of pulmonary tuberculosis diagnosed by fibreoptic bronchoscopy. AB - A total of 356 patients were subjected to fibreoptic bronchoscopy from September 1989 to June 1991 to exclude bronchial carcinoma. Bronchial biopsy, bronchial brush smears and bronchial wash were obtained. Bronchial wash was examined for acid fast bacilli (AFB) compatible with Mycobacterium tuberculosis. The total number diagnosed as pulmonary tuberculosis by fibreoptic bronchoscopy was 21(5.8%). The sputum smears were negative for AFB in all these patients. Previous studies have shown the importance of fibreoptic bronchoscopy in suspected cases of tuberculosis where the sputum smear is negative. This study is further evidence of the importance of routine examination of bronchial wash for AFB in all cases undergoing fibreoptic bronchoscopy to detect atypical cases of pulmonary tuberculosis. PMID- 8234106 TI - Naftidrofuryl treatment for rest cramp. AB - A double-blind placebo-controlled study was carried out in 14 subjects to investigate the effect of naftidrofuryl in the treatment of rest cramp. Naftidrofuryl caused a significant reduction in cramp frequency (median naftidrofuryl = 5; median placebo = 17; P < 0.004) and a significant 34% increase in cramp free days (median naftidrofuryl = 22; median placebo = 14; P < 0.004). Naftidrofuryl is an effective alternative to quinine in the treatment of this painful condition. PMID- 8234107 TI - Colon filling defect. PMID- 8234108 TI - Curriculum design for primary care physicians. PMID- 8234109 TI - The role of clinical tutors. PMID- 8234110 TI - Lymphocytic meningitis as the sole manifestation of Q fever. AB - A young man who presented with a 3 week history of fever and severe headache accompanied by mild leukocytosis, was found to have lymphocytic meningitis due to Coxiella burnetti. Thus, Q fever can present as lymphocytic (aseptic) meningitis responsive to tetracycline with no evidence of pulmonary involvement. PMID- 8234111 TI - Pseudotumour cerebri associated with arteriovenous malformations. AB - The association of intracranial hypertension and arteriovenous malformations is described in two patients. Both patients had the typical clinical features of pseudotumour cerebri and were found to have intracranial arteriovenous malformations on arteriography. The mechanism of raised intracranial pressure in patients with arteriovenous malformations is discussed. PMID- 8234112 TI - Salivary gland involvement as initial presentation of Wegener's disease. AB - We describe a case of Wegener's disease with massive painful bilateral submandibular gland enlargement as the presenting symptom. The diagnosis was based on histologically documented nasal mucosa involvement, focal necrotizing glomerulonephritis, episcleritis and anti-neutrophil cytoplasmic antibody. PMID- 8234113 TI - Acquired C1 inhibitor deficiency with lymphoma causing recurrent angioedema. AB - A 52 year old man who developed recurrent, massive and generalized angioedema for the first time during adult life was found to have an acquired deficiency of C1q esterase inhibitor (C1 INH) in association with a B cell lymphoma producing a paraprotein. He had low levels of C4 and C1 INH during the attacks which returned to normal after the successful treatment of lymphoma. An underlying lymphoproliferative disease should always be considered in adult patients with this immunological profile, recurrent angioedema and a negative family history. PMID- 8234114 TI - Malignant change in dermatitis artefacta. AB - Dermatitis artefacta is a chronic skin lesion produced by self-trauma. Avoidance of further trauma, topical steroids and psychological therapy all play a part in the treatment of such lesions. Unresolved lesions may become large and disfiguring and subject to infection. We report a case of one such lesion in an elderly woman who persistently excoriated a cholecystectomy scar over 40 years. Malignant transformation occurred in a manner analogous to the neoplastic change observed in other types of chronic ulcer (Marjolin's ulcer). The squamous cell carcinoma presented with widespread metastases from which the patient eventually died. Recent literature concerning Marjolin's ulcers is reviewed and it is noted that this is the first reported case of death caused by malignant change in dermatitis artefacta. PMID- 8234115 TI - Hairy leukaemic cell influx into the cerebrospinal fluid secondary to encephalomyelitis. AB - A case report of a 53 year old male with hairy cell leukaemia is presented in whom encephalomyelitis caused by toxoplasmosis resulted in an influx of hairy cells into the cerebrospinal fluid following disruption of the blood-brain barrier. These cells subsequently disappeared as the barrier reformed. It is suggested that the presence of hairy cells in the cerebrospinal fluid is a secondary self-limiting phenomenon. PMID- 8234116 TI - Haemolytic-uraemic syndrome with thrombotic microangiopathy of the retina following cytomegalovirus infection: postmortem findings. AB - Haemolytic-uraemic syndrome is characterized by thrombotic microangiopathy of the glomeruli and smaller arterial vessels of the kidney. Extrarenal thrombotic microangiopathy occurs, but ocular involvement is rarely demonstrated microscopically. We describe a 33 year old woman with a 3 week febrile episode and seropositivity for cytomegalovirus infection who developed acute renal failure, blindness and severe encephalopathy which was the cause of death. Thrombotic microangiopathy of retinal vessels may be more common than is expected from the literature. The exclusion of ocular structures in postmortem examinations explains the lack of anatomoclinical correlation. PMID- 8234117 TI - Sclerosing cholangitis associated with Crohn's disease and autoimmune haemolytic anaemia. AB - A middle-aged man was found to have autoimmune haemolytic anaemia. Seven years after the first manifestations of the anaemia, he developed jaundice without haemolysis and a diagnosis of primary sclerosing cholangitis was made by endoscopic retrograde cholangiography. Crohn's colitis was later confirmed by X rays and colonoscopy. This association is unique to the best of our knowledge and suggests that genetic and immunological mechanisms may be involved in the pathogenesis of these diseases. PMID- 8234118 TI - Gastric phytobezoar: unusual association and resolution. AB - A 58 year old man underwent a laparotomy for intestinal obstruction, which proved to be due to intestinal pseudo-obstruction. Following a slow postoperative course, he was discovered to have a gastric phytobezoar. The association between bezoars and pseudo-obstruction has not been previously described, but might be expected. The gastric phytobezoar could not be fragmented mechanically, but it completely disappeared when enteral feeding was commenced. The possibility of a future role of enteral feeding in this situation is suggested. PMID- 8234119 TI - Paracetamol cardiotoxicity. PMID- 8234120 TI - Maternal paroxysmal supraventricular tachycardia treated with adenosine. PMID- 8234121 TI - Correlated responses to selection for high or low incidence of tibial dyschondroplasia in broilers. AB - Selection for a high or a low incidence of tibial dyschondroplasia (TD) at 7 wk of age was practiced for four generations in broiler breeders keeping an unselected control (C) line and using low intensity X-ray imaging for TD diagnosis. The incidence of TD, BW at 4 and 7 wk of age, age at first egg, and egg production for the first 12 wk of production were determined in the four generations. Age at initial semen production and semen volume and concentration of males were determined only in Generations 3 and 4. The TD scores and BW at 7 wk were collected on 1,366, 1,301, and 1,389 broilers from the C, high (H), and low (L) incidence of TD lines. Incidence of TD at 7 wk in the fourth generation was 16.1, 61.9, and 3.5% for the C, H, and L lines, respectively. There were no significant differences in BW between the H and L lines in Generations 1, 3, or 4. However, in Generation 2, the H line was heavier (P < .01) than the L line. Second generation H line broilers with TD were heavier than those without TD at 4 and 7 wk of age (P < .01). There were no significant differences in age at first egg, age at semen production, or in semen volume between the H and the L lines. Egg production in the L line was greater (P < .01) than that of the H line in all four generations, and semen concentration was also greater (P < .01) in Generations 3 and 4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234122 TI - Responses of dwarf and normal chickens to feed restriction, Eimeria tenella infection, and sheep red blood cell antigen. AB - Relationships among stress responses, habituation to feed restriction, resistance to Eimeria tenella, and antibody response to SRBC were studied in dwarf and normal White Plymouth Rocks. Transfer of chicks at 22 days of age from starter to developer batteries resulted in an increase within 24 h of heterophil:lymphocyte (H:L) ratios of chicks of both genotypes. Restriction of feed intake from ad libitum (AL) to 60% of ad libitum reduced body weight and increased size of the crop-esophagus. As measured by H:L ratios, the effect of adapting to the 60% feed restriction dissipated between 12 and 16 days after initiation. Release of 60% restricted chicks to 80% of ad libitum also elicited a stress response as measured by H:L. These effects were noted in both dwarf and normal chicks. Time required for chicks on restriction to consume their daily allotment was curvilinear with a plateau occurring at a smaller value for dwarf than for normal chicks. Neither feeding regimen nor genotype had an effect on antibody response to SRBC. Resistance to E. tenella was greater in normal than dwarf chicks and greater for restricted than ad libitum chicks. PMID- 8234123 TI - The efficacy of using prelay and early-lay rations for brown-egg pullets. AB - Experiments were conducted using two brown-egg strains to determine whether additional calcium or CP is required by these pullets prior to sexual maturity (prelay) and whether increased CP during the early part of production (early lay) would improve overall performance. In Experiment 1, the prelay treatments were arranged at a 2 x 3 factorial arrangement with two levels of CP (15 or 17%) and three levels of calcium (.9, 1.8, or 3.5%) and given to the pullets from 18 to 20 wk of age. From 20 to 26 wk of age, pullets were given either a 17 or 19% CP ration with 3.5% calcium, resulting in a total of 12 treatments. There were no significant effects of the prelay or early-lay rations on eggs per hen housed, hen-day percentage production (HDP), or feed efficiency. Egg weight was significantly improved using the 17% prelay ration at 26, 28, and 31 wk of age, but adversely affected by the higher levels of CP in the early-lay ration at 28 and 31 wk of age. Increasing calcium in the prelay ration had only minimal effects on egg size distribution. Experiment 2 consisted of a 3 x 2 factorial arrangement of treatments with three levels of prelay CP (15, 17, or 19%) and two levels of early-lay CP (17 or 19%) and given to the pullets from 18 to 20 to 41 wk of age, respectively. There was a significant improvement in HDP from 30 to 41 wk of age using the 19% CP prelay ration. Egg size distribution was not significantly affected by either the prelay or the early-lay rations at 35 wk of age. PMID- 8234124 TI - Broiler chick and laying hen tolerance to sodium hypochlorite in drinking water. AB - Two broiler and two laying hen experiments were conducted to establish threshold levels for sodium hypochlorite use in poultry drinking water. Both broiler experiments were of 21-days duration, and the first and second hen experiments ran for 8 and 4 wk, respectively. A 5.25% sodium hypochlorite solution was used to provide chloride levels of 0, 10, 100, or 500 ppm in the first broiler study and 0, 150, 200, 250, 300, 350, 400, or 450 ppm in the second study. Comparable sodium levels from sodium bicarbonate were additional treatments in the first broiler experiment. The initial hen study had drinking water treatments of 0, 50, or 100 ppm chloride whereas in the second study, treatments varied from 0 to 70 ppm chloride at 10-ppm intervals. In chick experiments, water intake was reduced by 100 ppm chloride and body weight by 300 ppm. Hen experiments demonstrated the adverse effect level to be 40 ppm for water consumption and 60 ppm for egg production when the application was to pullets producing eggs at a high rate in warm weather. Water consumption of older hens in cooler weather was adversely affected by 50 ppm chloride, but egg production, egg weight, or daily feed intake was not reduced by 100 ppm. PMID- 8234125 TI - Feed intake adjustments by hens to feeding regimens in which dietary methionine is varied. AB - At 20 wk of age, 315 ISA Brown pullets were individually caged and randomly assigned to one of seven feeding regimens for the next 20 wk. In Regimen 1 Diet A was fed, in Regimen 2 Diet B was fed, Regimens 3 and 4 alternated Diets A and B weekly (the week Diet A was fed in Regimen 3, Diet B was fed in Regimen 4), Regimens 5 and 6 alternated Diets A and B monthly (reversing as with 3 and 4), and in Regimen 7 pullets were given a choice of Diets A and B. Diet A was methionine-deficient (.23%) and Diet B was balanced for methionine (.33%). Feed and nutrient consumption, egg production, egg weight, and egg mass were lower in Regimen 1 than 2. Pullets in Regimen 1 lost body weight and those in Regimen 2 gained weight. Overall feed consumption for Regimens 3 through 7 was similar to that for Regimen 2. For Regimens 3 through 6, however, consumption increased when the balanced diet was fed and decreased when the deficient diet was fed. Concomitant with these oscillations in feed consumption were changes in egg mass, so that means for these traits were intermediate and different from those for Regimens 1 and 2. For Regimen 7, in which Diets A and B were continuously available, no preference of diet was observed during the first 4 wk.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234126 TI - Two-step mucosal competitive exclusion flora treatment to diminish salmonellae in commercial broiler chickens. AB - There is a need to control the intestinal colonization of broiler chickens by salmonellae in order to reduce the contamination of poultry products. A two-step treatment of broiler chicks with a mucosal competitive exclusion culture (MCE) was tested, in which the MCE was first sprayed on chicks in the hatchery followed by administration in the first drinking water. Three commercial flocks were treated and compared with parallel, untreated control flocks. Customary husbandry practices were employed. Environmental, hatchery, skin with feathers, and cecal samples were analyzed at 3 and 7 wk for the presence of salmonellae. Carcass rinse samples of fully processed birds were analyzed similarly. The results indicated that initial feed, water, and litter contamination was at a low frequency (< 10%). Eggshell fragments and chick paper pads were frequently contaminated (> 50%). After 3 wk growth, contamination of litter, skin with feathers, and ceca were significantly (P < .05) reduced in treated flocks as compared with control flocks. Salmonellae prevalence in ceca and in processed carcass rinses was also significantly (P < .05) reduced from 41% in control flocks to 10% in treated flocks. The study showed that treatment of chickens in a commercial setting with MCE cultures can serve as a useful means to reduce salmonellae contamination. PMID- 8234127 TI - Effect of atherogenic diet on chicken plasma lipids and lipoproteins. AB - Adult male White Leghorn chickens were used in an experimental model system to study atherogenesis, and the effects of an atherogenic diet on plasma lipoprotein composition including carotenoids were determined. This model also included treatment with diazepam, a drug known to reduce formation of atherogenic plaques. After 6 wk consumption of a high-cholesterol, high-triglyceride diet, chickens had mean total plasma cholesterol, triglyceride, and carotenoid concentrations that were significantly increased over those from chicks that consumed the standard diet. Diazepam treatment had no significant effect on whole plasma concentrations of these lipids. Total body weight gain was unaffected by diet, but liver weight expressed as percentage of body weight was significantly increased in chickens that consumed the atherogenic diet. High density lipoprotein (HDL) and low density lipoprotein (LDL) fractions were isolated from plasma samples by ultracentrifugation. The atherogenic diet increased the carotenoid, cholesterol, and protein content of the LDL fractions but not the HDL fractions. PMID- 8234128 TI - Cutaneous hypersensitivity responses in chickens divergently selected for antibody responses to sheep red blood cells. AB - Cellular immunity in vivo of chickens selected for high and low antibody responses to SRBC was evaluated by primary and secondary cutaneous hypersensitivity reactions to BSA and phytohemagglutinin (PHA)-induced delayed type hypersensitivity (DTH). Similar to PHA-induced DTH, primary and secondary cutaneous hypersensitivity to BSA comprised acute (1- and 2-h), intermediate (4- and 6-h), and late (24-h) wing web swelling responses. Primary and secondary hypersensitivity to BSA revealed line differences between the High (H) line and a randombred Control (C) line on the one hand, and the Low (L) line on the other hand. Hypersensitivity to BSA was almost absent or decreased in the L line. The late phase of PHA-induced DTH was significantly elevated in the H line as compared with the L line. Naive recipients sensitized systematically with either serum from chickens sensitized with BSA, or with BSA-binding IgG, developed an antigen-specific early wing web swelling response following local challenge with BSA. It is suggested that the acute component of cutaneous hypersensitivity is mediated by Ig. The acute response may be a prerequisite for the development of classical (late) cutaneous DTH responses or may be initiated simultaneously with DTH. The current results indicate that the selection did not result in a higher hypersensitivity response to another T cell-dependent antigen and a mitogen in the H line, but in a lower response in the L line. PMID- 8234129 TI - Effects of selected minerals on acid-base balance and tibial dyschondroplasia in broiler chickens. AB - Experiments were conducted to determine the relative acidogenicities of several anions, including chloride, sulfate, and mono-, di-, and tribasic phosphate in commercial broiler chickens. Graded levels (80 and 160 meq/kg of diet in two experiments; 50, 100, and 150 meq/kg in a third experiment) of all anions were substituted as calcium salts on a molar equivalent basis for calcium carbonate in a semipurified diet. All diets contained equivalent amounts of calcium. The occurrence of tibial dyschondroplasia was determined by visual scoring of the epiphyseal growth plate in the distal end of the tibiotarsus. None of the anions added to the diet at levels of 160 meq/kg or less affected body weight or feed consumption. A higher level of chloride (240 meq/kg) that was utilized in some experiments decreased weight gain by 16 to 22%. All anions except tribasic phosphate resulted in reduced blood pH or bicarbonate concentrations or both. Monobasic phosphate was more acidogenic than dibasic phosphate and sulfate and chloride was the most acidogenic of all mineral anions. All anions including tribasic phosphate increased the severity of tibial dyschondroplasia. There was no consistent relationship between blood pH, bicarbonate concentration, or partial pressure of CO2 and the severity of tibial dyschondroplasia. PMID- 8234130 TI - Influence of prelay treatment and dietary protein level on the reproductive performance of White Leghorn hens. AB - Two experiments were undertaken to study the influence of prelay diet treatment on subsequent laying performance. Feeding either a conventional laying or growing diet or diets containing essentially only wheat bran from 18 to 21 wk resulted in little grower diet carryover effect beyond 24 wk of age, although the wheat bran diets resulted in a marked drop in body weight of the pullets during the prelay treatment. Similar performance was noted for pullets in a second experiment in which prelay treatments consisted of feeding just corn or wheat bran from 18 to 20 wk of age. In interpreting the results of the present study, it is important to consider the fact that "mature pullet weights" were obtained before the prelay diet treatments were employed. Where prelay diet treatments have had significant effects on laying house performance, in most cases, immature or underweight pullets were involved. Feeding hens low-protein diets (13 versus 17%) from 20 to 44 wk of age resulted in similar egg production; however, egg weight and thus egg mass were slightly reduced with the lower protein diet. There was no laying by growing treatment interaction. The importance of good pullet weight at the start of the production cycle and the subsequent performance of such pullets fed low protein diets is discussed. PMID- 8234131 TI - Jojoba meal (Simmondsia chinensis) in the diet of broiler breeder pullets: physiological and endocrinological effects. AB - The present studies evaluated the ability of jojoba meal (JO) to inhibit feed intake of broiler breeder pullets to limit body weight gain as recommended by the breeder company. A first experiment, using graded levels of JO supplementation (0 to 12%), was conducted to establish appropriate JO supplementation. Adequate reduction of growth rate was obtained with 4% JO supplementation. However, notwithstanding their similar growth rate, 4% JO chickens consumed considerably more feed compared with feed-restricted chickens. The dose-dependent impairment of feed intake with increasing levels of JO supplementation was also associated with increased plasma growth hormone and thyroxine and with decreased plasma insulin-like growth factor-I and triiodothyronine concentrations compared with 0% JO chickens. A second experiment included a pair-fed group. Notwithstanding their similar feed intake, 4% JO chickens gained significantly less body weight compared with their pair-fed counterparts. The 4% JO chickens also had a longer feed transit time per kilogram body weight. Again, circulating levels of the somatotrophic and thyrotrophic hormones were altered according to the dietary treatment. From all these observations, it was concluded that the growth retardation caused by JO supplementation was provoked by an inhibition of appetite linked with the simmondsin content of JO as well as by other antinutritional compounds affecting digestibility. PMID- 8234132 TI - The influence of differences in dietary amino acids during the early growing period on growth and development of Nicholas and British United Turkey toms. AB - From 0 to 4, 4 to 8, and 8 to 12 wk of age, Nicholas (NIC) and British United Turkeys (BUT) toms were fed one of two series of experimental diets. All toms were fed the same diet, appropriately adjusted for age period, from 12 to 16 and 16 to 24 wk. The first series was formulated to 1984 NRC specifications whereas the second series contained higher concentrations of lysine and total sulfur amino acids (NRC+). The NIC toms were heavier (P < or = .05) at 4, 8, 12, and 16 wk, but there were no strain differences at 20 wk. The NRC+ diets significantly increased BW in both strains at 12 wk, but there were no diet effects at 16 or 20 wk. At 12 wk, BUT toms had significantly more carcass protein and less carcass lipid compared with NIC toms, but there were no significant dietary effects on these variables. In both strains, the NRC+ diets significantly increased the relative weight of the eviscerated carcass and both the absolute and relative weights of the Pectoralis major muscles at 12 wk. At 20 wk, the absolute and relative weight of the Pectoralis minor muscles was significantly heavier in BUT toms and in toms fed the NRC+ diets. The tibia and femur were longer in BUT toms compared with NIC toms. From 1 to 24 wk, weekly BW from 50 toms per strain and treatment combination were used to calculate growth curve parameters according to the Gompertz equation. The slope of the growth curve was significantly less in BUT toms, but the BW and age at the point of infection were increased compared with NIC toms. There were no significant diet effects on any growth curve parameters. PMID- 8234133 TI - Effect of enzyme supplementation on the performance and digestive tract size of broiler chickens fed wheat- and barley-based diets. AB - Three experiments were conducted to study the effects of crude enzyme preparations on the performance and gastrointestinal tract size of chicks fed wheat and barley diets. In the first experiment, enzyme addition (100 and 200 mg/kg of Roxazyme G and 1,000 mg/kg of Avizyme SX) to diets containing Bedford barley improved weight gain (6%) and the feed to gain ratio (5%) over a 6-wk period for both male and female broilers. In Experiment 2, enzyme addition to diets containing Scout (hulless) and Bedford (hulled) barley improved (P < or = .05) weight gains of Leghorn chicks by 25 and 11% and the feed to gain ratios by 10 and 6%, respectively. Feed consumption increased significantly (16%) only in the case of birds fed enzyme with Scout barley. Corresponding reductions in the relative weights of the crop and gizzards were 15 and 17% for birds fed Scout barley and 7 and 8% for those fed Bedford barley. Enzyme treatment of the diet containing Scout barley also reduced the relative length of the duodenum, jejunum, and ileum and the relative weight of the proventriculus, whereas a similar treatment of Bedford barley resulted in changes in the relative length of the duodenum and jejunum (P < .05). In the final broiler experiment (42 days), crude enzyme addition (100 mg/kg) to wheat and barley diets improved weight gains by 13 and 9% and feed to gain ratios by 7 and 10%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234134 TI - Evaluation of egg injection method of pantothenic acid in turkey eggs and effect of supplemental pantothenic acid on hatchability. AB - Two experiments were conducted with a commercial strain cross of 120 Large White British United Turkeys of America to determine the effect of pantothenic acid egg injections and dietary pantothenic acid on hatchability. The hens were housed individually in cages in a conventional house. In Experiment 1, three dietary treatments were used: 1) an unsupplemented practical corn-soybean meal basal diet; 2) the basal diet supplemented with 37.4 mg pantothenic acid/kg; and 3) the basal diet supplemented with 74.8 mg pantothenic acid/kg. Incremental dietary supplemental pantothenic acid levels increased the transfer of pantothenic acid in eggs, but did not result in a hatchability increase over the unsupplemented pantothenic acid basal diet. The response patterns from dietary pantothenic acid for the reproductive variables were similar whether the data were analyzed on a production period basis using all of the hens or on a subset of hens producing eggs in each production period. In Experiment 2, with hens fed 37.4 mg supplemental pantothenic acid/kg of diet, hatchability did not increase in eggs injected with 1,800 micrograms pantothenic acid per egg as compared with uninjected eggs and eggs injected with the vitamin carrier solution. The results of the study indicate that hatchability was not increased in turkey eggs from hens fed supplemental pantothenic acid or with egg pantothenic acid injections, which suggests that pantothenic acid is not limiting for hatchability in commercial turkey hen diets that contain 10.5 mg/kg or more of pantothenic acid. PMID- 8234135 TI - Estimates of copper bioavailability from liver of different animal species and from feed ingredients derived from plants and animals. AB - Bile Cu accumulation in Cu-depleted chicks fed Cu concentrations between .56 and 1.56 mg/kg was used to estimate Cu bioavailability in several feed ingredients from both plant and animal sources, including liver from different species. Liver from slaughtered animals is rich in minerals and vitamins and is a significant source of Cu in canned pet foods. Liver from different species, however, was found to vary widely in Cu bioavailability (relative to CuSO4.5H2O, which was set at 100%). The bioavailability of Cu in freeze-dried (FD) chicken liver and poultry by-product meal was 116 and 97%, respectively, but that in FD pork liver was not different from zero. Relative bioavailability of Cu in FD beef, sheep, and turkey liver was 82, 113, and 83%, respectively. Copper in FD liver from the rat, a species that does not possess a gall bladder, was 21% bioavailable. Copper in the feed ingredients from plants: corn gluten meal, dehulled soybean meal, cottonseed meal, peanut hulls, and soy mill run was 48, 38, 41, 44, and 47% bioavailable, respectively. In addition, when the fibrous ingredients peanut hulls or soy mill run were added to the basal diet containing .5 mg Cu/kg from CuSO4.5H2O, Cu bioavailability in CuSO4.5H2O was reduced. The results of this study demonstrate a wide variation in Cu bioavailability among feed ingredients originating from plants and animals. PMID- 8234136 TI - Median eminence and anterior pituitary degradation of luteinizing hormone releasing hormone in hens undergoing changes in luteinizing hormone secretion. AB - Studies described in this report provide physiological evidence for a possible involvement of median eminence (ME) and anterior pituitary (AP) luteinizing hormone releasing hormone degrading activity (LHRH-DA) in the genesis of the hen's preovulatory surge of luteinizing hormone (LH). Serum LH and progesterone (P4), ME LHRH content and LHRH-DA, and AP LHRH-DA were determined in laying hens in the following reproductive conditions: 1) a "spontaneous" preovulatory LH surge; 2) a "premature" preovulatory LH surge; and 3) an ovulatory failure induced by feed withdrawal. The premature preovulatory surge of LH occurred 3 h after P4 administration and was preceded by an increase in both ME and AP LHRH-DA and by a decrease in ME LHRH content 1 h after P4 administration. However, the premature preovulatory LH surge was associated with a decrease in LHRH-DA back to control levels as ME LHRH content increased, even though LHRH was presumably being released from the ME at this time to maintain the preovulatory surge of LH. Although similar changes in LHRH-DA (an increase in both ME and AP LHRH-DA, followed by a decrease) also preceded the spontaneous preovulatory surge of LH, its profile was significantly blunted and no changes in ME LHRH content were associated with this LH surge. In contrast, ovulatory failure was correlated with a decrease in ME LHRH content but no changes in LHRH-DA. Therefore, in the hen, ME and AP enzymatic degradation of LHRH I might be involved in the genesis of a premature preovulatory surge of LH. PMID- 8234137 TI - Perinatal thyroid hormones and hepatic 5'deiodinase in relation to hatching time in weight-selected lines of chickens. AB - Plasma thyroid hormone concentrations, liver weight, body weight, and hepatic capability for triiodothyronine (T3) production were measured at 8-h intervals on Days 19 through 21 in embryos and chicks from lines selected for high (HW) and low (LW) body weight at 8 wk of age. Embryos were categorized as not having entered the perinatal period (nonpipped, NP) or as being in one of three perinatal stages: embryos that had pipped into the air cell (AC), embryos pipped through the shell (TS), and chicks within 24 h of hatching (H). Both plasma thyroxine (T4) and T3 peaked at the TS stage in embryos of both lines. Embryos had higher concentrations of both hormones and hatched earlier in the LW than in the HW line. This association and evidence from other studies is suggestive that T3 may be playing a role in line differences in hatching time. Hepatic 5'deiodinase (5'D) activity was higher in LW than HW embryos and chicks at the AC, TS, and H stages although the opposite had been the case prior to the perinatal period (NP stage). Hepatic 5'D increased at each stage from AC to TS to H. When the patterns of liver growth and body weight were considered, this pattern and the differences between the lines were exaggerated further. Comparison of the plasma and 5'D patterns during the perinatal period suggests that substrate (T4) supply may be more important than 5'D capability of the liver in determining the pattern of T3 supply during some of these stages. PMID- 8234138 TI - Fate of Listeria monocytogenes and pediococcal starter cultures during the manufacture of chicken summer sausage. AB - Two formulations of chicken summer sausages [100% hand deboned chicken meat (HDCM) and 85% HDCM and 15% chicken hearts (HDCM-CH)] were prepared with a nonpediocin-producing (PED-) Pediococcus acidilactici starter culture and inoculated with 10(4) or 10(7) cfu of a five-strain mixture of Listeria monocytogenes/g of batter. Sausages were fermented to pH 5.0 (11 h), cooked to an internal temperature of 66.5 C, cold-showered, and stored at 4 C (60 days) and 30 C (7 days). For both formulations and inoculation levels, L. monocytogenes populations decreased 1.3 to 1.8 log10 cfu/g by the end of fermentation. No L. monocytogenes organisms were recovered from sausages (by enrichment) following the cook and shower or storage at 4 or 30 C. In contrast, P. acidilactici increased .7 to 1.2 log10 cfu/g during fermentation, and < 10(2) cfu/g remained after the cook and shower and storage at 4 and 30 C. In a second set of experiments, sausages (HDCM) were prepared with a PED- or a pediocin-producing (PED+) P. acidilactici starter culture and challenged with the L. monocytogenes mixture (10(7) cfu/g). The PED- culture reduced numbers of L. monocytogenes 1.2 log10 cfu/g during fermentation, whereas L. monocytogenes numbers declined 2.6 log10 cfu/g in the presence of the PED+ culture. Although acid production by both starter cultures was equivalent, greater inhibition of L. monocytogenes by the PED+ compared with the PED- starter culture was attributed to in situ production of pediocin. Pediococcal starter cultures and proper cooking eliminated L. monocytogenes from sausages and established that PED+ cultures provide an additional hurdle against poultry-related listeriosis. PMID- 8234139 TI - Research note: the effect of organic acids on Salmonella contamination originating from mouse fecal pellets. AB - Laboratory mice were inoculated with a nalidixic acid and novobiocin-resistant strain of Salmonella typhimurium. Contaminated fecal pellets were harvested 2 days postinoculation. Each half of a fecal pellet was found to contain equal numbers of Salmonella on a per weight basis. When separate halves were placed into a poultry feed, either treated or untreated, with an organic acid mixture (SAL CURB), the treatment was able to significantly (P < .001) reduce Salmonella contamination of the feed by almost two log orders per gram when compared with the untreated control. PMID- 8234140 TI - Research note: effect of carbonated drinking water on production performance and bone characteristics of laying hens exposed to high environmental temperatures. AB - An experiment was conducted to determine the effects of providing heat-stressed laying hens with carbonated drinking water on production performance and tibia bone breaking strength. A total of 32 commercial laying hens (89 wk of age) were housed in an environmental chamber and exposed to 1 wk of a constant thermoneutral temperature (21 C), followed by 6 wk of a daily heat stress temperature cycle (34 C for 8 h; 21 C for 16 h). One half of the hens received tap water and the other half were provided carbonated water immediately after housing. Following 6 wk of heat stress temperature exposure, all hens were killed by cervical dislocation and left tibia bone strength measurements were taken. Hen day egg production, egg weight, egg yield, feed consumption, and feed efficiency did not differ between water treatments; however, egg specific gravity was depressed for hens provided carbonated versus tap drinking water. Left tibia bone breaking strength per 100 g body weight was improved (P < or = .05) for hens provided with carbonated drinking water during exposure to heat stress temperatures. These results suggest that carbonated drinking water may enhance bone integrity by increasing tibia bone breaking strength of older laying hens exposed to a short-term heat stress period. PMID- 8234141 TI - Therapeutic potential of cholecystokinin receptor antagonists in CNS disorders. PMID- 8234142 TI - Favourable effects of sub-MIC rufloxacin concentrations in decreasing the pathogen-host cell adhesion. AB - The in vitro effects of sub-inhibitory concentrations of rufloxacin on Escherichia coli and Staphylococcus aureus adherence to human buccal epithelial cells were investigated. Epithelial cells were incubated with bacteria, grown in presence of serial dilutions of rufloxacin, from 1/2 MIC (Minimum Inhibitory Concentration) to 1/128 MIC. The mean number of bacteria adhering to cells and the percentage of cells bearing > or = 50 bacteria on their surface were counted and compared with those of controls (cells incubated with bacteria without rufloxacin). For both the bacterial strains tested, the maximal inhibition of bacterial adhesion occurred at 1/2 MIC concentrations. A lower, but statistically significant, inhibition was also observed with other sub-MIC concentrations up to 1/32 for both S. aureus and E. coli adhesion. These findings indicate that sub inhibitory concentrations of rufloxacin, which may be present in vivo in the mucosae in the troughs of the dosage cycle, significantly inhibit the adhesive properties of bacteria involved in mucosal colonization. PMID- 8234143 TI - Possibility of spontaneous drug abuse tested in rat. AB - Given that a number of the techniques used to test drug abuse liability are not free from criticism, a series of oral free-choice experimental procedures was adopted. When simultaneously offered as alternatives to glucose using the classical polydipsic procedure, no preference for buprenorphine (0.025 mg/ml), morphine (0.5 mg/ml) or fentanyl (0.005 mg/ml) solutions was shown by premedicated rats. The same result was obtained when the two-bottle procedure was used for at least one month to offer etonitazene (10 micrograms/ml), buprenorphine (60 micrograms/ml), cocaine (300 micrograms/ml) and haloperidol (25 micrograms/ml) solutions as simultaneous alternatives to aspartame. This absence of preference was maintained even when the rats showed evident pharmacological effects and, in the case of the opiates, tolerance and withdrawal syndrome. However, when a gustatory marker (quinine) was introduced into one of the two solutions, preference was always shown for the other. Finally, in a conditioned taste aversion (CTA) test, etonitazene (5 or 40 micrograms/kg, i.p.) and haloperidol (0.5 or 2 mg/kg, i.p.) did not induce any reduction in saccharin consumption, while morphine (40 mg/kg) did. Pretreatment with naloxone (120 micrograms/kg, i.c.v.) did not antagonize morphine-induced CTA, while it did antagonize morphine-induced analgesia. PMID- 8234144 TI - No modifications of GABAA and benzodiazepine receptors following experimental dysthyroidism in rats. AB - The effect of a treatment with L-triiodothyronine (T3) or propylthiouracil (PTU) on the characteristics of benzodiazepine and chloride ion channel binding sites in rat hippocampus and cerebral cortex was studied using a radiolabelled technique. In our experimental conditions, neither hyper- nor hypothyroidism modified number and affinity of [3H]flunitrazepam or [3H]butylbicycloorthobenzoate (TBOB) binding sites. These data indicate that neither benzodiazepine nor chloride ionophore sites of the GABA complex are modified in an experimental condition of dysthyroidism. PMID- 8234145 TI - Flumazenil prevents the anxiolytic effects of diazepam, alprazolam and adinazolam on the early acquisition of two-way active avoidance. AB - The effects of diazepam (DZ, 4 mg/kg), alprazolam (ALP, 1.25 mg/kg) and adinazolam (ADIN, 6 mg/kg), as well as their interaction with the benzodiazepine receptor antagonist flumazenil (Ro15-1788), were studied on the early acquisition of two-way active (shuttlebox) avoidance in male Sprague-Dawley rats. The three benzodiazepines increased shuttlebox avoidance acquisition, and their effects were prevented (antagonized) by flumazenil (10 mg/kg). The present results indicate that central benzodiazepine (BZ) receptors are involved in the anxiolytic effects of diazepam and triazolobenzodiazepines on the early acquisition of two-way active avoidance. PMID- 8234146 TI - Effect of a benzylidene derivative, a novel antirheumatic agent, on IL-1 production. AB - 3,4-Diacetoxy benzylidene diacetate (ACP) is a prodrug of protocatechualdehyde (PAL). PAL significantly inhibited interleukin-1 (IL-1) production and release in human monocytes in a dose dependent fashion under lipopolysaccharide (LPS) stimulation. ACP showed inhibitory effects on cartilage destruction of the femoral condyles induced by adjuvant arthritis in vivo in a significant and dose dependent fashion. To clarify the mechanism of action of ACP on rat adjuvant arthritis, we investigated the effects of PAL, a metabolite of ACP, on IL-1 production using synovial cell cultures derived from patients with rheumatoid arthritis. PAL significantly inhibited the IL-1 beta production induced by IL-1 alpha or PMA without inhibition of total protein synthesis and cytotoxicity. A protein kinase C (PKC) inhibitor, staurosporine, also suppressed the IL-1 beta production induced by IL-1 alpha or PMA, suggesting that the PKC pathway plays an important role in IL-1 alpha-induced IL-1 beta production. The calcium ionophore A23187 (A23187) potentiated the IL-1 beta production induced by IL-1 alpha. Whereas PAL slightly inhibited under these conditions, it was not statistically significant. These results suggest that PAL has a favourable action on cartilage destruction through the inhibition of IL-1 production induced by the modification of the PKC pathway. PMID- 8234147 TI - Efficient production of the cyclosporin metabolite AM1 by rabbit hepatic enzymes. AB - We describe here conditions for the in vitro preparation of the main metabolite of cyclosporin A, namely AM1, with rabbit liver enzymes. The compound was purified and was identified by comparison with the metabolite isolated from urine of rats treated with [3H]CsA and by its Mass-Spectrometry Fast Ion Bombardment (MS-FIB) and 1HNMR spectra. The procedure is simple and with the enzyme fraction derived from one rabbit liver 3-4 mg of the pure metabolite can be obtained. PMID- 8234148 TI - The Association of State and Territorial Directors of Nursing: leadership in official state agencies. PMID- 8234149 TI - One hundred years of powerful women: a conversation with Myrtis J. Snowden. Interview by Ann H. Cary. PMID- 8234150 TI - One hundred years of powerful women: a conversation with Sylvia R. Peabody. Interview by Grace P. Erickson. PMID- 8234151 TI - Sense of coherence in minority women at risk for HIV infection. AB - It is theorized that persons with strong sense of coherence are likely to define an event as less stressful and be able to manage a problem more successfully than those with weak sense of coherence. The study investigated the relationship among coherence and personal and environmental concerns, appraisal of threat, emotional distress, and high-risk behaviors in minority women at risk for human immunodeficiency (HIV) infection. As predicted, a significant negative relationship was seen between level of coherence and concerns. Moreover, women strong in coherence reported less negative appraisals of threat, less emotional distress, and fewer high-risk behaviors than those with weak coherence. Results of tests of a path model investigating the impact of coherence and appraisal on distress and risk revealed coherence to be significantly and negatively associated with appraisal, distress, and risk, both directly and indirectly through its association with appraisal. The path model accounted for 45% of the variance in distress, 10% of the variance in appraisal, and 4% of risk behavior. Continuing investigation of factors such as coherence that can lessen the seriousness of environmental stressors is imperative as it relates to women at risk for HIV infection. PMID- 8234152 TI - Knowledge of AIDS and safer sex practices among college freshmen. AB - We assessed knowledge of the acquired immunodeficiency syndrome (AIDS) and of safer sex practices among college freshmen. A second purpose of the study was to assess this knowledge among black as well as white students. Students attending classes at three private colleges in a large southern city were asked to participate in the study. Respondents completed the modified AIDS information survey, the knowledge of safe sex practices questionnaire, and a demographic data sheet. A total of 689 questionnaires were received from single college freshmen. The results indicated that respondents were knowledgeable about the cause and transmission of AIDS but were less knowledgeable about medical aspects. Most knew that condoms are effective in preventing the spread of AIDS, but fewer could differentiate between the effectiveness of latex and nonlatex condoms. These findings are useful to health educators in improving AIDS education programs. PMID- 8234153 TI - A model of health care delivery in a child day-care setting. AB - Many children in our country spend all or part of their days in group day care while their parents work or go to school. These children are not only at risk for day-care-related illnesses, but also may lack preventive health care because their parents have time constraints or financial barriers. A group of nursing faculty members found that by collaborating with early childhood educators, accessible and cost-effective health care could be provided to children at a campus day-care site. This collaboration allowed for the early detection of problems, thereby reducing barriers to learning. Furthermore, by entering a day care system, nurses found opportunities to conduct health education and promotion that had a positive impact on children, parents, and day-care staff. This model, or components of it, could be replicated in similar settings. PMID- 8234154 TI - Five hundred cases of pneumonia. 1902. PMID- 8234155 TI - Realities of developing community health orientation programs. AB - The realities of developing an orientation program include recognizing the critical influence of organizational culture and management priorities on the final form of plans and projects. The step from theory to practice is illustrated in the experience of three of the authors as they began to implement competency based orientation programs. The common experiences of these three very different agencies were that programs are affected by organizational readiness, evolution to meet organizational needs, and the administrative context. PMID- 8234156 TI - Theory-based practice: attitudes of nursing managers before and after educational sessions. AB - In Canada, professional associations are mandating theory-based nursing as a standard of practice. Few systematic studies of nurses' attitudes toward, knowledge of, and exposure to theory-based nursing are in the literature. We used a convenience sample of nursing managers in a public health department to describe these aspects before and after an educational program, and to identify preferred ways of learning about nursing models. The managers had moderately positive attitudes toward theory-based nursing, which increased and were maintained six months after an educational program. Case studies and small group discussions were preferred over lectures and reading as methods of learning nursing theory. The significant increases in positive attitudes after the educational program suggest that knowledge and experience in applying nursing theory had an impact on the managers' attitudes. Managers identified a need to explore a variety of models before making a commitment to one. Leadership, commitment, and careful planning are required when the decision to introduce a model is made. PMID- 8234157 TI - Predicting use of hearing protection: a preliminary study. AB - The National Institute of Occupational Safety and Health targeted noise-induced hearing loss as one of the top 10 occupational hazards. Research has been directed toward reducing noise exposure rather than identifying the factors associated with workers' decision consistently to use hearing protection. The long-term goal of this program of research is to develop a causal model explaining workers' use of hearing protection to guide development of nursing interventions. This study was a preliminary one to prepare for testing Pender's health-promotion model (HPM) as a causal model. Ninety-eight skilled tradesmen completed written questionnaires at their work site. Although 98% indicated they were supposed to wear protection, their reported use averaged 50.3% of the time. Reported use significantly and positively correlated with workers' perceptions of the benefits and self-efficacy of use, value of outcomes regarding keeping out noise and increased well-being, and health-promoting behaviors in the areas of self-actualization and stress management. Perceptions of barriers of use were significantly and negatively related to use. In regression analyses of use of hearing protection on components of the HPM and dimensions of health behavior and lifestyle, one component of the HPM (benefits) and two dimensions of health behavior and lifestyle (self-actualization and interpersonal support) significantly predicted use, accounting for 24% of the variance in use of protection. Knowledge of the predictors of hearing protection use will ultimately aid nurses in implementing interventions, increasing use, and decreasing hearing losses. PMID- 8234158 TI - Illicit drug use among young adults with genital herpes. AB - Although an association exists between illicit drug use and sexually transmitted diseases (STDs) in known high-risk populations such as prostitutes, gay men, and intravenous drug users, little is known with respect to middle-class, heterosexual populations thought to be at less risk. We examined findings related to illicit drug use among young adults with genital herpes from a larger study of young adults' adaptation to chronic disease. In this cross-sectional retrospective study, 70 subjects age 18 to 35 years were interviewed and asked to fill out questionnaires. The sample was predominantly white, heterosexual, employed, and college educated. Responses to open-ended questions related to illicit drug use were coded and compared with findings from two other surveys: a national household survey, Pacific region, metropolitan areas; and a household survey of a San Francisco Bay area county. All but eight subjects (88.6%) reported that they had used an illicit drug at some time in the past. The median number of drugs used was 4; the maximum number was 11. Only six drug-using subjects reported that they had restricted their use to marijuana. Those with genital herpes were more likely to report ever using an illicit drug at some time in the past, and, more specifically, in the past year, than subjects in the two comparison surveys. Such findings suggest that programs screen for illicit drug use as a risk factor in middle-class young adults with genital herpes and other STDs, and for STDs in persons referred for drug abuse treatment. PMID- 8234159 TI - The relation of public health nursing to the public health campaign. 1915. PMID- 8234160 TI - The use of solution theories for predicting water vapor absorption by amorphous pharmaceutical solids: a test of the Flory-Huggins and Vrentas models. AB - The limitations of traditional gas adsorption models for describing water vapor sorption by amorphous pharmaceutical solids are described and an alternative approach based on polymer solution theories is proposed. The approach is tested by comparing a priori predicted isotherms with literature data for the poly(vinylpyrrolidone)(PVP)-water system. The well-known Flory-Huggins model is able to describe the water vapor sorption isotherm only when the PVP-water mixture is in the rubbery state (i.e., above its glass transition temperature). However, a newer model developed by Vrentas and coworkers, which takes into account the plasticizing effect of water on the polymer, is able to describe the entire form of the isotherm. Consideration of the parameters in this model allows a number of critical variables to be identified and also enables the characteristic shape of the water vapor sorption isotherm to be explained. PMID- 8234161 TI - Peptide stability in drug development. II. Effect of single amino acid substitution and glycosylation on peptide reactivity in human serum. AB - The determination of peptide stability in human serum (HS) or plasma constitutes a powerful screening assay for eliminating unstable peptides from further development. Herein we report on the stability in HS of several major histocompatibility complex (MHC)-binding peptides. Some of these peptides are in development for the novel treatment of selected autoimmune disorders such as rheumatoid arthritis and insulin-dependent diabetes. For most of the l-amino acid peptides studied, the predominant degradation mechanism is exopeptidase-catalyzed cleavage. Peptides that were protected by d-amino acids at both termini were found to be more stable than predicted, based on additivity of single substitutions. In addition, N-acetylglucosamine glycopeptides were significantly stabilized, even when the glycosylation site was several amino acids from the predominant site(s) of cleavage. This indicates that long-range stabilization is possible, and likely due to altered peptide conformation. Finally, the effect of single amino acid substitutions on peptide stability in HS was determined using a model set of poly-Ala peptides which were protected from exopeptidase cleavage, allowing the study of endopeptidase cleavage pathways. PMID- 8234162 TI - Preparation, characterization and pharmaceutical application of linear dextrins. I. Preparation and characterization of amylodextrin, metastable amylodextrins, and metastable amylose. AB - The linear dextrin amylodextrin was prepared by enzymatic hydrolysis from waxy maize. Four metastable amylodextrins were prepared by complexation with different volatile organic compounds. All products showed partial dissolution into water at room temperature, because of dissolution of molecules with a lower DP. X-ray diffractometry revealed a helical conformation with six glucose units per turn for amylodextrin and metastable amylodextrins prepared with small molecules, and a helical conformation with seven glucose units per turn for metastable amylodextrins prepared with larger molecules. All metastable amylodextrins showed a helix with reduced distance between two turns as compared to amylodextrin. Metastable amylose, prepared from Amylose V, showed a helical conformation again with a reduced distance between two turns compared to Amylose V. FTIR analysis indicated a more flexible conformation for Amylose V and metastable amylose than for the amylodextrins. PMID- 8234163 TI - Preparation, characterization, and pharmaceutical application of linear dextrins. II. Complexation and dispersion of drugs with amylodextrin by freeze-drying and kneading. AB - The ability of amylodextrin (a linear dextrin) to act as a complexing agent or as a carrier for solid dispersion was evaluated. Blends of amylodextrin with diazepam or prednisolone were freeze-dried and kneaded at elevated temperatures, respectively. The products were analyzed by DSC, X-ray diffractometry, and FTIR spectroscopy. Complex formation with amylodextrin by freeze-drying was found not to occur for diazepam but for prednisolone at a molar ratio of 1 to 1. The freeze dried product of diazepam with amylodextrin proved to be a solid dispersion. Solid dispersions were formed by both wet (with ethanol) and dry kneading at elevated temperatures of low-melting drugs such as lidocain, diazepam, and methyl PABA with amylodextrin. No solid dispersions were obtained for high-melting drugs such as prednisolone and salicylic acid. The results point to the formation of solid dispersions by a melting mechanism during the process of kneading at elevated temperatures of low-melting drugs with amylodextrin. PMID- 8234164 TI - Evaluation of the physicochemical properties and dissolution characteristics of mesalamine: relevance to controlled intestinal drug delivery. AB - The physicochemical properties of mesalamine and the effect of pH and buffer concentration on the dissolution rate of pure mesalamine and mesalamine with Carbopol 974P were investigated. The aqueous solubilities at 25 and 37 degrees C were 0.844 and 1.41 mg/mL, respectively. Consistent with the observed pKa1 (2.30) and pKa2 (5.69) or mesalamine, the solubility-pH profile is increased at pH < 2.0 and pH > 5.5 and is minimized from pH 2.0 to pH 5.5. The flux data were consistent with the solubility data from pH 1.0 to pH 5.5. The flux increased and plateaued at pH values 5.5 to 7.0 and was dependent on the bulk buffer concentration. At low bulk buffer concentrations, mesalamine reduces the pH in the diffusion layer, which results in a decrease in flux. The medium with the highest buffer capacity has a greater ability to increase the surface pH and dissolution rate. The addition of Carbopol reduces the flux and the sensitivity of the dissolution rate of mesalamine to increasing bulk buffer concentration. This reduction is postulated to be due to neutralization of the basic dissolution media, gel formation, and possible drug-polymer interactions. PMID- 8234165 TI - In vivo skin penetration of acitretin in volunteers using three sampling techniques. AB - Etretinate and acitretin are given orally to treat psoriasis and various keratinization disorders. Acitretin, the main active metabolite of etretinate, has the pharmacokinetic advantage of being rapidly eliminated, but it shares etretinate's toxicologic profile. Thus a topical delivery of acitretin with no or reduced systemic adverse effects is desirable. To characterize the therapeutic potential of topically delivered acitretin, we quantitatively assessed its percutaneous penetration in healthy human volunteers. Additionally, three skin sampling techniques, the punch biopsy, the shave biopsy, and the suction blister technique, were validated to quantitate acitretin in the skin. The results suggest that topical delivery of acitretin renders skin concentrations which exceed those reported after oral administration of etretinate or acitretin. However, because of possible interlaminate drug contamination, drug localization within a particular skin compartment cannot be determined. PMID- 8234166 TI - The dissolution and bioavailability of etodolac from capsules exposed to conditions of high relative humidity and temperatures. AB - The dissolution and bioavailability of etodolac from capsules exposed to high relative humidity and temperature were compared to those from capsules stored at room temperature (RT). Dissolution of stressed and control capsules was evaluated using a USP basket apparatus at 100 rpm with 900 mL pH 7.5 phosphate buffer (0.05 M) at 37 degrees C. The dissolution of etodolac from capsules exposed to stressed conditions was also evaluated with enzymes (pancreatin, 1%, w/v) added to the dissolution medium. The bioavailability of etodolac from capsules exposed to stressed conditions was compared in both dogs and humans to capsules stored at RT conditions. Capsules, 200 and 300 mg, exposed to stressed conditions failed the dissolution (without enzymes) specification [not less than 85% released (80% Q) in 30 min]. However, upon enzyme addition, all capsules met the specification. The rate and extent of absorption from these 200 and 300 mg etodolac capsules in dogs were equivalent to those from capsules stored at RT conditions that passed the dissolution specification. Similarly, the bioavailability of etodolac from 300 mg capsules that failed the dissolution specification upon exposure to stressed conditions was equivalent to that of control capsules in 24 adult male volunteers. Thus, an in vitro dissolution test with enzymes provides a better indication of stressed capsule performance in vivo. PMID- 8234167 TI - Lack of effect of ammonium glycyrrhizinate on the morphology of ovine nasal mucosa in vitro. AB - Glycyrrhetinic acid derivatives are reported to be nasal absorption promoters (1). Effects of ammonium glycyrrhizinate (AMGZ) on the in vitro morphology of ovine nasal mucosa were therefore examined by light and electron microscopy. Nasal mucosa was stripped from the submucosa and mounted in Ussing chambers. Exposure of the apical surface to 2% ammonium glycyrrhizinate (24 mM) for 90 min caused no histopathological changes to the nasal epithelium. Epithelial integrity remained intact as evidenced by the continued presence of morphologically intact junctional complexes. No sloughing of the epithelial layer from the basement membrane was observed, and cilia and microvilli were not affected by treatment with AMGZ. The results indicate that short-term exposure in vitro to ammonium glycyrrhizinate caused no overt morphological damage to ovine nasal mucosa. PMID- 8234168 TI - Dissolution modeling: factors affecting the dissolution rates of polydisperse powders. AB - The dissolution rates of two lots of hydrocortisone (fine and coarse) were simulated using a computer program based on a Noyes-Whitney-type equation. Derivations of the equation were made to compare the accuracy of simulations using spherical and cylindrical particle geometry, with and without a time dependent diffusion layer thickness. To approximate better the shape of the hydrocortisone particles, a shape factor was used to relate cylindrical length to radius. The most accurate simulations were obtained by assuming cylindrical geometry with and without a time-dependent diffusion layer thickness for the fine and coarse hydrocortisone, respectively. The program was also modified to simulate initial particle size distributions based on the log normal probability density function. PMID- 8234169 TI - Convective solvent flow across the skin during iontophoresis. AB - Enhanced flux of neutral solutes during transdermal iontophoresis is attributed largely to electroosmotic volume flow. In this study, the iontophoretic fluxes of tritiated water (3H2O) and 14C-labeled mannitol through hairless mouse skin (HMS) were determined. The following questions were addressed: (i) What is the variability of water flux during iontophoresis? (ii) To what extent is the iontophoretic flux of a neutral solute correlated with water flux? (iii) Does the intrinsic permeability of the skin to neutral solutes change following iontophoresis? (iv) What is the effect of low pH on electroosmotic volume flow? and (v) Does the skin remain permselective after removal of the stratum corneum? Transport of both water and mannitol reached steady-state levels during 10 hr of constant-current iontophoresis (0.36 mA/cm2). Anodal fluxes exceeded cathodal values. Cathodal mannitol flux was retarded, relative to passive transport, by net volume flow in the opposite direction, such that transport of this molecule increased significantly after the termination of current passage. Anodal equivalent volume flows for water and mannitol, respectively, were 2.7 (+/- 1.3) and 1.23 (+/- 0.59) microL/hr cm2, indicating that only approximately 50% of the water flux participated in the electroosmosis of mannitol. The passive permeability of water and mannitol after 10 hr of iontophoresis was, respectively, 6 and 30 times greater than the pretreatment values. At pH 7, the cationic permselectivity of HMS was marginal [the Na+ transport number (tNa+) was determined to be 0.46] and less than that reported for human skin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234170 TI - A floating controlled-release drug delivery system: in vitro-in vivo evaluation. AB - A novel floating controlled-release drug delivery system was formulated in an effort increase the gastric retention time of the dosage form and to control drug release. The buoyancy was attributed to air and oil entrapped in the agar gel network. A floating controlled-release 300-mg theophylline tablet having a density of 0.67 was prepared and compared in vitro and in vivo to Theo-dur. The in vitro release rate of the floating tablet was slower. In vivo scintigraphic studies for a floating and a heavy nonfloating tablet, under fasting and nonfasting conditions, showed that the presence of food significantly increased the gastric retention time for both tablets, and tablet density did not appear to make a difference in the gastric retention time. However, the positions of the floating and nonfloating tablets in the stomach were very different. Bioavailability studies in human volunteers under both fasting and nonfasting conditions showed results comparable to those with Theo-dur. The floating controlled-release theophylline tablet maintained constant theophylline levels of about 2 mg/mL for 24 hr, which may be attributable to the release from the agar gel matrix and the buoyancy of the tablet in the stomach. PMID- 8234171 TI - Topical penetration of piroxicam is dependent on the distribution of the local cutaneous vasculature. AB - The mechanism of the topical delivery of piroxicam, a nonsteroidal antiinflammatory drug, has been controversial as to whether systemic absorption is required for topical efficacy. This study, using in vivo pigs treated with topical 3H-piroxicam gel, was designed to assess the role of systemic absorption on its delivery to deep tissues. Further, the role of the structure of the cutaneous vasculature (e.g., direct cutaneous or musculocutaneous) was studied. Finally, piroxicam delivery was measured using in vitro diffusion cells with pig skin obtained from the same sites to determine inherent permeability independent of vascular anatomy. These studies showed that penetration of the radiolabel occurred in subcutaneous and muscle tissue only under the dosed sites and not at the remote sites, ruling out systemic absorption as a prerequisite for local delivery. Tissue penetration in vivo was enhanced at the musculocutaneous compared to the direct cutaneous sites. In contrast, in vitro flux was identical in skin harvested from the two vascular sites, suggesting that the vasculature plays a pivotal role in deep tissue penetration of piroxicam. In conclusion, local delivery of topical drugs occurs independent of systemic absorption and the nature of the cutaneous vasculature at different sites must be taken into consideration for optimal delivery. PMID- 8234172 TI - Rates of systemic degradation and reticuloendothelial system (RES) uptake of thermosensitive liposome encapsulating cisplatin in rats. AB - The systemic degradation and reticuloendothelial system (RES) uptake of cisplatin (CDDP)-encapsulated thermosensitive liposomes composed of dipalmitoylphosphatidylcholine (DPPC) and distearoylphosphatidylcholine (DSPC) (DPPC/DSPC = 9/1, 7/3, and 5/5, w/w) after intravenous administration to rats were examined by measuring the platinum (Pt) levels in the blood and RES (liver and spleen). The blood liposome level profile showed first-order rate elimination for each liposome administration. The elimination rate (Kel) was faster when the content of DSPC was lower (Kel: 1.3/hr for 9/1-liposomes, 0.7/hr for 7/3 liposomes, 0.5/hr for 5/5-liposomes). On the other hand, the RES liposome level profile showed distribution of liposomes followed by elimination therefrom. The RES level of the liposomes was lower when the content of DSPC was smaller (maximal level: 25% for 9/1-liposomes at 1 hr, 32% for 7/3-liposomes at 1 hr, 37% for 5/5-liposomes at 2 hr). The kinetic analysis demonstrated that the RES uptake rate (Kres) was almost the same among the liposomes (0.4/hr), while the systemic degradation rate (Kdeg; Kel-Kres) became larger as the content of DSPC decreased (0.9/hr for 9/1-liposomes, 0.3/hr for 7/3-liposomes, and 0.1/hr for 5/5 liposomes) and that the RES liposome distribution amount was dependent not only on the Kres but also on the Kdeg and the rate of RES liposome degradation. The Kdeg for each type of liposome corresponded with the systemic CDDP release rate. PMID- 8234173 TI - Effect of age on distribution of zidovudine (azidothymidine) into the cerebrospinal fluid of Macaca nemestrina. AB - The brain tissue is an important target for anti-HIV drug therapy. Since the permeability of the blood-brain and blood-cerebrospinal fluid (CSF) barriers may differ between neonates and adults, we have determined the effect of age on the distribution of zidovudine (ZDV or azidothymidine) into the CSF in the macaque (M. nemestrina). Five newborn macaques were administered ZDV (iv bolus, 5 mg/kg) at various ages (2 days to 4 months). Both CSF (cisternal) and venous blood samples were obtained at approximately 60 and 90 min after drug administration. In another series of experiments, adult female macaques received ZDV as either an iv bolus (5 and 10 mg/kg) or an infusion for at least 12 hr. CSF (lumbar) and venous blood samples were obtained at approximately 60 and 90 min after iv bolus and at more than 12 hr after iv infusion. ZDV concentration in the CSF and the plasma samples was determined by high-performance liquid chromatography. The CSF/plasma concentration ratio of ZDV in the newborn and adult macaques, after iv bolus administration, was independent of time. In addition, no significant (P > 0.05) difference was observed in the pooled iv bolus ZDV CSF/plasma concentration ratio between the adult group (0.236 +/- 0.058) and the newborns (0.213 +/- 0.039). Moreover, the ZDV CSF/plasma concentration ratio in the adults and the newborns, after iv bolus administration, was found not to be significantly (P > 0.05) different from the ratio obtained at steady state in the adults (0.224 +/- 0.094). These data indicate that the distribution of ZDV into the CSF in macaque neonates and adults is similar. PMID- 8234174 TI - Effect of apparent elimination half-life on nitroglycerin-induced hemodynamic rebound in experimental heart failure. AB - Hemodynamic rebound after abrupt withdrawal may be an important consideration associated with nitroglycerin (NTG) monotherapy. This phenomenon may arise from unopposed neurohormonal vasoconstriction because of rapid elimination of NTG. The role of NTG pharmacokinetics in the development of hemodynamic rebound was examined using a rat model of congestive heart failure. NTG was infused for 90 min, then the dose was either abruptly stopped (n = 8) or gradually reduced by 20% every 20 min (n = 7). Abrupt withdrawal caused rebound elevations of left ventricular end-diastolic pressure (LVEDP) to about 25% above baseline values, at 30-60 min after drug termination (P < 0.01), but this was completely avoided by graded NTG withdrawal. A positive correlation was observed (P < 0.05) between the percentage reduction in LVEDP during infusion and the maximum percentage rebound in rats after abrupt withdrawal but not after graded withdrawal. These results suggest that NTG-induced hemodynamic rebound is related to its short biological half-life and that this phenomenon is consistent with a mechanism of neurohormonal compensation. PMID- 8234175 TI - Correlation between the plasma concentration of mepirodipine and its occupancy of Ca2+ antagonist receptors in rats. AB - The relationship between the plasma concentration of mepirodipine (1,4 dihydropyridine Ca2+ antagonist) and its occupancy of cardiac and cerebral Ca2+ antagonist receptors in rats has been characterized by a radioreceptor assay technique using (+)-[3H]PN 200-110. Oral administration of mepirodipine in rats produced a dose-dependent and sustained decrease in the number of specific (+) [3H]PN 200-110 binding sites in both tissues, and the effect was more pronounced in the cardiac tissue than in the cerebral cortex. The occupancy of cardiac and cerebral Ca2+ antagonist receptors by mepirodipine correlated well with its plasma concentration, whereas a 20-fold higher plasma concentration of this drug was necessary to occupy Ca2+ antagonist receptors in the cerebral cortex. Thus, these data suggest that mepirodipine occupies Ca2+ antagonist receptors in cardiovascular tissue selectively over those in brain tissue. PMID- 8234176 TI - Phosphoryloxymethyl carbamates and carbonates--novel water-soluble prodrugs for amines and hindered alcohols. AB - Phosphoryloxymethyl carbonates and carbamates of the type R1R2X-CO-O-CH2-O-PO3( 2) (X = O or N) were evaluated as potentially novel water-soluble collapsible prodrugs for alcohols and amines. These were prepared by reaction of alpha chloromethyl chloroformate with the starting alcohol or amine to give the corresponding alpha-chloromethyl carbonate or carbamate, respectively. Reaction with silver dibenzyl phosphate followed by debenzylation by hydrogenolysis gave the desired products. The aqueous chemical stability of the phosphoryloxymethylcarbonyl derivatives of 2-indanol (3a), beta-(3,4 dimethoxyphenyl)ethylamine (3b), and benzocaine (3c) were evaluated. The aqueous hydrolysis of 3a-3c resulted in regeneration of the parent alcohol or amines. As expected, the hydrolytic behaviors of these derivatives were found to differ from that of simple alkyl and aryl phosphomonoesters. The rates of hydrolysis were extremely rapid, with the dianionic phosphate species possessing a higher reactivity than the monoanionic species. This was attributed to the proximity of the phosphate group to the carbonyl moiety. The carbamate derivatives, 3b and 3c, displayed greater chemical stability compared to the carbonate derivative, 3a. Alkaline phosphatases-mediated hydrolysis of the phosphate ester bond in 3c led to a rapid cascade reaction resulting in regeneration of the parent amine, benzocaine. Although the alcohol derivative described here appeared to be too chemically unstable to be ideal as a prodrug, the derivatives of the amines might have some use. They are expected to be cleaved in vivo by alkaline phosphatases. PMID- 8234177 TI - The effect of dihydronicotinate N-substitution on the brain-targeting efficacy of a zidovudine chemical delivery system. AB - Enhanced brain delivery of zidovudine (AZT) has been demonstrated using a redox based chemical delivery system (CDS). Optimization of the prototype AZT-CDS (5' [(1-methyl-1,4-dihydropyridin-3-yl)carbonyl]-3'-azido-3'-deoxy thymidine ) was investigated by manipulation of the N-methyl group present on the dihydronicotinate portion of the molecule and examining the release of AZT in vivo in a rat model. Of the five compounds examined, all produced higher brain levels and lower blood levels of AZT than did AZT itself. In comparing the novel AZT-CDS analogues to the N-methyl benchmark, the N-propyl system proved to be the most efficient of the compounds tested. PMID- 8234178 TI - The effect of swelling characteristics of superdisintegrants on the aqueous coating solution penetration into the tablet matrix during the film coating process. AB - The mechanism and the extent of sorption of water molecules by tablets containing superdisintegrants and microcrystalline cellulose, following the aqueous film coating of formulated tablets, were investigated. The penetration of water from the coating solution into the tablet matrix resulted in significant changes in physical properties of the coated tablet cores, such as residual moisture content, tensile strength, and pore-size distribution. The swelling and the morphological characteristics of each individual disintegrant compound and microcrystalline cellulose were found to have important implications on the extent of penetration of water from the aqueous film coating solution. A hypothesis concerning the interaction between microcrystalline cellulose and the superdisintegrant particles present in the tablet matrix is proposed. PMID- 8234179 TI - The influence of blood sampling site in nasal drug delivery studies in rats. PMID- 8234180 TI - Single-puff particle-size analysis of albuterol metered-dose inhalers (MDIs) by high-pressure liquid chromatography with electrochemical detection (HPLC-EC). PMID- 8234181 TI - Preparation of small gelatin and albumin microparticles by a carbon dioxide atomization process. PMID- 8234182 TI - Solid-state emulsions: the effects of maltodextrin on microcrystalline aging. PMID- 8234183 TI - BRL 38227--a potassium channel opener, antagonizes digoxin-induced convulsions. AB - Experimental evidence suggests that potassium channel openers play an important role in convulsions. In this study, the anticonvulsant activity of BRL 38227, a new potassium channel opener against digoxin-induced convulsions, is reported. Intraventricular administration of digoxin (7.5 micrograms), included "popcorn type" convulsions in rats. BRL 38227, injected centrally increased the onset time of convulsions and decreased the mortality rate in a dose-dependent manner. Pretreatment with 4-aminopyridine, a potassium channel blocker antagonized the protective effect of BRL 38227. These findings show the involvement of potassium channels in digoxin-induced convulsions. Further these results indicate that in the future potassium channels might be a target for new anticonvulsant drugs. PMID- 8234184 TI - Oxysterols inhibit gap junctional communication between rat hepatocytes in primary culture. AB - Several oxysterols were examined for their effect on gap junctional communication between rat hepatocytes in primary culture. 25-Hydroxycholesterol, 22(S) hydroxycholesterol and 7 beta-hydroxycholesterol, in decreasing order of potency, markedly inhibited gap junctional communication. In contrast, 7-ketocholesterol showed no inhibitory effect. The inhibition of gap junctional communication by oxysterols was not a consequence of changes in cell viability, as measured by lactate dehydrogenase leakage and 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) reduction activity. The addition of exogenous cholesterol to the culture medium did not abolish the effect of 25 hydroxycholesterol, suggesting that the capacity of oxysterols to inhibit gap junctional communication is independent of their inhibitory effect on cholesterol synthesis. We suppose that inhibition of gap junctional communication may be an early sign of oxysterols-induced toxicity on hepatocytes. PMID- 8234185 TI - Active transport of benzylpenicillin across the blood-milk barrier. AB - Passage of benzylpenicillin across the mammary gland epithelium was studied both after systemic administration of benzylpenicillin in four goats and after intramammary infusion in three goats and two cows. The results after benzylpenicillin administration alone were compared with the results after combined administration of benzylpenicillin and probenecid. The studies on passage of benzylpenicillin through the blood-milk barrier showed, when milk to plasma ultrafiltrate ratios of benzylpenicillin were plotted versus time for each half of the udders, that active transport takes place from blood to milk during steady-state plasma concentrations. Active transport was demonstrated by inhibition with probenecid: (1) The entry of benzylpenicillin into milk was slowed down under the influence of probenecid. The AUC (area under the curve) values during the first 30 min. were reduced by 66 +/- 9% compared with the AUC values found without coadministration of probenecid. (2) In the presence of probenecid and during equilibrium between blood and milk concentrations, benzylpenicillin reached concentration ratios in the ultrafiltrates of milk and plasma which corresponded to those expected for diffusion alone. Without probenecid these ultrafiltrate concentration ratios were more than two times higher indicating an active transport of benzylpenicillin from blood to milk. After intramammary infusion of benzylpenicillin, active transport was demonstrated from milk into blood. The absorption rate for benzylpenicillin from the mammary gland was reduced by probenecid, as measured by the ratio of benzylpenicillin to urea absorption half-life, which was increased by 40-50% in the presence of probenecid. PMID- 8234186 TI - Prenatal toxicity of xylene inhalation in the rat: a teratogenicity and postnatal study. AB - Technical xylene (cas. nr. 1330-20-7) was investigated for development toxicity in a teratology and in a postnatal study. Rats (Mol: WIST) were exposed to 500 p.p.m. 6 hr per day on days 4 to 20 of gestation. There were no signs of maternal toxicity. In the teratology study, no exposure-related differences were found except for delayed ossification of os maxillare. In the postnatal study, the xylene-exposed pups had a higher body weight and an impaired performance on a motor ability test (Rotarod). Due to the possibility of direct toxic effects of xylene on the developing central nervous system, further studies are needed to investigate dose-effect relationship for this type of effects. PMID- 8234187 TI - 15-Deoxyspergualin prolongs pancreatic islet allo- and xenograft survival in mice. AB - The new immunosuppressant 15-deoxyspergualin was evaluated in allogeneic and xenogeneic pancreatic islet transplantation. In the allograft study 500 collagenase-isolated C57BL/6 mouse islets were transplanted under the renal capsule of alloxan-diabetic C57BL/Ks mice that were either 15-deoxyspergualin treated (n = 15) or given saline only (n = 8). When 15-deoxyspergualin was given (5 mg/kg b.wt. intraperitoneally) until day 28 after transplantation in a special dosage schedule, 10 out of 15 animals were normoglycaemic one week after transplantation and 6 were still normoglycaemic after ten weeks. All 8 control animals were hyperglycaemic after 18 days. Light microscopy showed graft rejection in hyperglycaemic mice, but only mild infiltration of lymphocytes in the grafts of normoglycaemic animals. In the xenograft study C57BL/Ks mice were transplanted under the renal capsule with 500-750 foetal porcine islet-like cell clusters. The grafts were examined for evidence of rejection with light microscopy at different time points after implantation. In the control animals given saline only (n = 37) there was progressive evidence of rejection starting on day seven. In 15-deoxyspergualin treated animals (2.5 mg/kg intraperitoneally; n = 27) there was significantly less infiltration at days 7, 14 and 21. After 32 days there was, however, no difference between controls and 15-deoxyspergualin treated mice. A doubling of the 15-deoxyspergualin dose (5.0 mg/kg intraperitoneally; n = 5) did not further improve the survival of the xenografted islet-like cell clusters. There was no synergistic effect when cyclosporine A (12.5 mg/kg intraperitoneally) was added to the 15-deoxyspergualin therapy (n = 34).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234188 TI - Some biochemical and pathophysiological aspects of long-term elevation of brain ornithine concentrations. AB - Mice and chicken were given 5-fluoromethylornithine (5FMOrn), a selective inactivator of ornithine aminotransferase (OAT) over extended periods of time. This treatment allowed us to maintain elevated concentrations of ornithine in all tissues. Since gyrate atrophy, an autosomal recessive human disease, is characterized by the absence of OAT, special emphasis was put on the study of the visual system. Ophthalmoscopic and histologic examinations of the eye as well as electroretinograms and locomotor behaviour demonstrated an unimpaired visual system and brain. No toxic effects were observable in the treated mice. Likewise, chick embryo development was normal in spite of highly elevated brain and tissue ornithine concentrations. A likely explanation for the absence of toxic effects of 5FMOrn treatment, disregarding the non-toxicity of ornithine, is the fact that 10-20% of tissue OAT is refractory to inactivation by 5FMOrn. This residual activity may be sufficient to maintain vital functions. PMID- 8234189 TI - Calcium antagonistic properties of the sesquiterpene T-cadinol and related substances: structure-activity studies. AB - The calcium antagonistic properties of (+)-T-cadinol, some of its stereoisomers and related terpenes were investigated in both functional and radioligand binding studies, and the effects were compared with those of the dihydropyridine calcium antagonist (+/-)-nimodipine. In the isolated rat aorta, the terpenes relaxed contractions induced by 60 mM K+ more potently than those induced by phenylephrine. (+)-T-cadinol and its stereoisomers were the most potent among the terpenes to relax K(+)-induced contractions, whereas they were approximately 10,000 times less potent than (+/-)-nimodipine in this regard. Binding of the dihydropyridine radioligand [3H]-(+)-PN200-110 was studied on rat cerebral cortical membranes. Displacement and saturation studies indicated that (+)-T cadinol caused a competitive inhibition of binding. The log Ki values for (+)-T cadinol and (+/-)-nimodipine from displacement studies (-4.7 and -9.2) corresponded with the log RC50 values for relaxation of K(+)-contracted rat aortas (-5.0 and -9.0). For the terpenes, there was a significant correlation (P < 0.001, rs = 0.89) between displacement of dihydropyridine binding and the ability to relax K(+)-induced contractions. The structures of three terpenes were chemically modified by blocking hydroxyl groups. The potency of these derivatives, as well as the naturally occurring derivative-2-oxo-T-cadinol, to relax K(+)-induced contractions was not correlated to the lipophilicity of the compounds. Instead, other qualities appear to be of importance for the functional effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234190 TI - Ethylmorphine metabolism in isolated rat hepatocytes. AB - The metabolism of ethylmorphine has been studied in suspensions of isolated rat hepatocytes. Early during incubation, the two major metabolic intermediates detected were morphine and norethylmorphine following N- and O-dealkylation of ethylmorphine, respectively. During subsequent incubation the concentration of the second metabolic intermediate, normorphine increased, before the concentration peaked at approximately 20 microM (100 microM ethylmorphine). Both morphine and normorphine were glucuronidated to form morphine-3-glucuronide and normorphine-3-glucuronide, respectively, which appeared to be the major metabolic end products. The percentage of ethylmorphine metabolized to morphine-3 glucuronide was found to be dependent on the initial concentration of ethylmorphine. With increasing initial ethylmorphine concentration the relative formation of morphine-3-glucuronide was reduced (29 +/- 10% at 5 microM, 18 +/- 5% at 20 microM, and 15 +/- 4% at 100 microM mean +/- S.D., n = 10). The concentrations of ethylmorphine and its metabolites were found to be higher in liver cells than in medium. Thus the ratios between the intra-/extra-cellular concentrations of ethylmorphine increased somewhat from an initial value of 4 during the period for which ethylmorphine could be detected intracellularly. The drug metabolites all exhibited ratios above 10 for the initial 100 min. of incubation. With time these ratios showed a decline, but even for prolonged incubation the ratios were 5 or higher for the end products. Thus considerable drug concentration gradients existed across the cell membrane of isolated rat hepatocytes. PMID- 8234191 TI - Inhibition of human neutrophil actin polymerization, phagocytosis and oxidative burst by components of decongestive nosedrops. AB - Human neutrophil functions have been examined after exposure of leukocytes to components of decongestive nosedrops in vitro. Both the vasoactive components oxymethazoline chloride and xylomethazoline chloride, as well as the preservative benzalkonium chloride, showed a concentration- and time-dependent deleterious effect on neutrophil actin polymerization, phagocytosis and oxidative burst. The most toxic of the drug components was benzalkonium chloride, which in the commercial nosedrops tested was present in concentrations about 20 times higher than that compatible with intact neutrophil functions. These findings suggest possible inhibition of mucosal neutrophil activity following exposure to nosedrops in vivo, and support earlier reports that have questioned the use of preservatives in decongestive nosedrops. PMID- 8234192 TI - Reproducibility over time of mephenytoin and debrisoquine hydroxylation phenotypes. AB - Mephenytoin and debrisoquine hydroxylation phenotypes were determined twice in 15 Spanish healthy volunteers with an interval of about one year. The phenotype assignment did not change in any subject for either debrisoquine or mephenytoin. Among extensive metabolisers of mephenytoin, there was a slight increase (P = 0.04) of the mephenytoin-S/R enantiomeric ratio over the study period. The family members of a poor metaboliser of mephenytoin were phenotyped, and the heterozygous extensive metabolisers were found to have higher mephenytoin-S/R ratios than other extensive metabolisers suggesting a correlation between the genotype and the S/R ratio. PMID- 8234193 TI - Reverse cholesterol transport in the rat: influence of short-term D-myo-inositol 1,2,6 trisphosphate (PP56) infusion. AB - D-myo-inositol-1,2,6 trisphosphate (PP56) was infused in rats for 3 hr at a rate of 30.28 microM/hr/kg body weight. The cholesterol content in plasma, erythrocytes, the aorta, and the liver was monitored before and at the end of the infusion and at 1 and 21 hr after the infusion. We found that the aortic total and free cholesterol content was reduced after the infusion and continuously until the end of the experiment (by 27% each after 21 hr P < 0.05, P < 0.01, respectively). Erythrocyte cholesterol was increased by 6.6% (P < 0.05) at the end of the infusion and 9.5% (P < 0.01) at the end of the experiment, while plasma and high-density lipoprotein cholesterol did not change significantly. Free cholesterol in the liver after infusion was decreased by 27% (P < 0.001) and cholesterol ester increased by 97% (P < 0.05) and did not return to the preinfusion level. Short-term PP56 infusion in the rat apparently influences cholesterol transport in a way which would suggest a promoted reverse cholesterol transport. Since the lipid values did not return to the preinfusion level, a long term study is needed to elucidate in more detail the influence of PP56 on cholesterol metabolism in vivo. PMID- 8234194 TI - Comparative acute lethality of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 1,2,3,7,8-pentachlorodibenzo-p-dioxin and 1,2,3,4,7,8-hexachlorodibenzo-p-dioxin in the most TCDD-susceptible and the most TCDD-resistant rat strain. AB - We have previously demonstrated a more than 300-fold difference in acute LD50 values for 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD) between male Long-Evans (Turku AB; L-E) and Han/Wistar (Kuopio; H/W) rats after intraperitoneal exposure. In the present study, we compared the acute lethality of TCDD, 1,2,3,7,8 pentachlorodibenzo-p-dioxin (PCDD) and 1,2,3,4,7,8-hexachlorodibenzo-p-dioxin (HCDD) in these strains by intragastric administration. In agreement with previous data, H/W rats proved to be strikingly resistant to TCDD, since even the highest dose tested, 7200 micrograms/kg, was below the LD50 level for both genders. The corresponding LD50 values for female and male L-E rats were 9.8 and 17.7 micrograms/kg, respectively. A similar strain difference was discovered for PCDD: the LD50 value was > 1620 micrograms/kg for female H/W rats and between 20 and 60 micrograms/kg for female L-E rats. Surprisingly, the acute lethality of HCDD did not follow the same pattern. Female H/W rats turned out to be only about 10 times less susceptible to that congener than female L-E rats (LD50 values 1871 and between 120 and 360 micrograms/kg, respectively). These findings do not support the widely accepted concept that sufficiently high doses of all dioxin congeners will produce the same effects. Either the higher chlorinated dioxins have toxic effects distinct from those of TCDD or the relative contribution of toxic impacts varies among these compounds. PMID- 8234195 TI - Important role of C-terminal peptide bonds for endothelin receptor binding and activity. PMID- 8234196 TI - Melanogenesis and the generation of cytotoxic molecules during insect cellular immune reactions. PMID- 8234197 TI - Interaction between chemicals and melanin. AB - Various drugs and other chemicals, such as organic amines, metals, polycyclic aromatic hydrocarbons, etc., are bound to melanin and retained in pigmented tissues for long periods. The physiological significance of the binding is not evident, but it has been suggested that the melanin protects the pigmented cells and adjacent tissues by adsorbing potentially harmful substances, which then are slowly released in nontoxic concentrations. Long-term exposure, on the other hand, may build up high levels of noxious chemicals, stored on the melanin, which ultimately may cause degeneration in the melanin-containing cells, and secondary lesions in surrounding tissues. In the eye, e.g., and in the inner ear, the pigmented cells are located close to the receptor cells, and melanin binding may be an important factor in the development of some ocular and inner ear lesions. In the brain, neuromelanin is present in nerve cells in the extrapyramidal system, and the melanin affinity of certain neurotoxic agents may be involved in the development of parkinsonism, and possibly tardive dyskinesia. In recent years, various carcinogenic compounds have been found to accumulate selectively in the pigment cells of experimental animals, and there are many indications of a connection between the melanin affinity of these agents and the induction of malignant melanoma. PMID- 8234198 TI - Free radicals in melanin-cationic porphyrin complexes in the dark and under light irradiation. AB - Electron Paramagnetic Resonance (EPR) spectroscopy was employed in the study of the interaction between L-3,4-dihydroxyphenylalanine (L-Dopa) melanin and the cationic porphyrins meso-tetrakis(1-methylpyridinium-4yl)-porphyrin (TMPyP), meso tetrakis-(1-benzylpyridinium-4-yl)-porphyrin (TBzPyP), and their respectives complexes ZnTMPyP and ZnTBzPyP. By monitoring signal intensities and progressive microwave power saturation it was shown that the interaction increases the equilibrium concentration of free radicals in L-Dopa melanin in the dark. The extent of increase is dependent on the presence of molecular oxygen and on the type of porphyrin. Not all interacting sites available for complexation in L-Dopa melanin are involved in the formation of free radicals. It was also observed that the interaction with porphyrins promotes an increase in the number of photoinduced free radicals in L-Dopa melanin during illumination with visible light. PMID- 8234199 TI - Application of photoacoustic phase angle spectroscopy (phi AS) to eumelanins and pheomelanins. AB - A new technique, based on the measurement of the phase angle of the photoacoustic signal, was used for investigating the absorption spectra of melanins in the dry state. The main advantages of such a method are the insensitivity to scattering of the light and the applicability to nontransparent or highly adsorbing and thermally thick samples. The spectra obtained on different kinds of natural and synthetic melanins show much more details than the corresponding ones obtained spectrophotometrically on aqueous suspensions of the pigment. Some relevant features of the spectra still cannot be interpreted, yet we were able to show the presence in eumelanins of dopachrome at 475 nm and melanochrome at 540 nm, and in pheomelanins of compounds previously isolated from red chicken feathers. Moreover, the well known trend of the optical absorption, i.e., its decrease with increasing wavelength, was unambiguously confirmed. PMID- 8234200 TI - Smyth chicken melanocyte autoantibodies: cross-species recognition, in vivo binding, and plasma membrane reactivity of the antiserum. AB - Smyth line (SL) chickens, which develop a depigmenting disorder similar to human vitiligo, produce circulating anti-melanocyte antibodies (Austin, L.M. et al., (1992) The detection of melanocyte autoantibodies in the Smyth chicken model for vitiligo. Clin. Immunol. Immunopathol., 64:112-120). In order to characterize these autoantibodies, we studied the reactivity of cultured chicken, mouse, and human melanocytes, as well as frozen sections of chicken feather follicles and embryonic eyes, against SL serum, employing indirect immunofluorescence. Light Brown Leghorn (LBL) serum was used as a negative control. Chicken (SL and LBL), mouse, and human melanocytes exhibited greater fluorescence with SL serum than with LBL serum (up to a 1:60,000 dilution). The fluorescent pattern was predominant along the perimeter of the cells, suggesting plasma membrane staining. Fluorescence-activated flow cytometry analysis and immunocytochemical localization at the ultrastructural level using intact chicken cells supported this hypothesis. Melanocytes were readily stained in cryosections of regenerating feather follicles and embryonic eyes incubated with SL, but not LBL, serum. In addition, amelanotic melanocytes in albino chicken feathers reacted with SL serum. SL serum also preferentially stained cells emigrating from cultured avian neural tubes and within the dermis of the proliferative germ of regenerating feather follicles suggesting that melanoblasts express the antigens. We conclude that Smyth line serum contains melanocyte autoantibodies that cross-react with mouse and human melanocytes, are able to bind to pigment cells within tissues, and recognize antigens expressed in the cytoplasm and on the surface of melanocytes and melanoblasts. PMID- 8234201 TI - Preparation of purified tyrosinase devoid of dopachrome tautomerase from mammalian malignant melanocytes. AB - Although tyrosinase has been considered for a long time the only enzyme involved in mammalian melanosynthesis, it has been shown that mouse melanoma melanosomes contain high levels of dopachrome tautomerase (DCT2), an enzyme catalyzing DC tautomerization to DHICA. At least in B16 mouse melanoma, DCT is present in higher catalytic amounts than tyrosinase. Moreover, it can be anticipated that tyrosinase and DCT should be very difficult to resolve by most conventional biochemical techniques because of the structural similarity between these enzymes, as predicted from the sequence of their corresponding cDNAs. It is shown that the presence of DCT can cause serious artifacts when tyrosinase activity is determined by most of the currently available methods, such as the Dopa oxidase and melanin formation assays. We describe a simple and convenient method for the preparation of tyrosinase devoid of DCT. The method takes advantage of the different thermal stability of both enzymes. Heating of crude melanosomal extracts at 60 degrees C for 1 hr results in a complete denaturation of DCT, while tyrosinase activity is recovered almost quantitatively. The resulting tyrosinase preparation is considerably purified and the electrophoretic, immunologic and kinetic characteristics of the enzyme appear unaltered. Because if its high yield and simplicity, the method can be used for the microscale partial purification of DCT-free tyrosinase from mammalian malignant melanocytes grown in culture. PMID- 8234202 TI - Chemical and electron microscopic studies of cattle (Bos taurus) with four types of phenotypic pigmentation. AB - The biological behavior of the pigmentary phenotypes of four breeds of cattle has been analysed: the black pigmentation of Holstein Friesian; the red pigmentation of Limousin; the dilution in Charolais; and the postnatal disappearance of red pigmentation in Chianina. The analytic techniques included the characterization of melanins by high-performance liquid chromatography, the examination of follicular melanocytes by light microscopy, and the examination of melanosomes by electron microscopy. The black phenotype was very strongly eumelanogenic. The red phenotype in Limousin is polymorphic: individual follicular melanocytes contain both mature eumelanosomes and pheomelanosomes. Charolais and Chianina cattle exhibited a dramatic reduction in melanogenic activity, which was characterized by the almost exclusive presence of prephaoemelanosomes in Charolais and of immature premelanosomes in Chianina. In the dilute Charolais phenotype, the density of distribution of follicular melanocytes also seemed to be reduced. The genes that are responsible for these four phenotypes seem to act on the maturation, differentiation, and density of distribution of the melanosomes. PMID- 8234203 TI - Evolution of the tyrosinase related gene (TYRL) in primates. AB - Tyrosinase is the major enzyme responsible for the formation of melanin pigment and is found throughout the animal kingdom. In humans, the tyrosinase gene (TYR) maps to the long arm of chromosome 11 at band q14-->q21, while a tyrosinase related gene (TYRL) maps to the short arm of chromosome 11 at p11.2-->cen. We and others have found that the TYRL locus contains sequences that are similar to exons IV and V of the authentic tyrosinase gene but lacks sequences of exons I, II, and III. In an attempt to understand the evolution of the human tyrosinase gene, we have analyzed TYR and TYRL in primates and have found that exons IV and V of the chimpanzee and gorilla TYR are very similar to the human, with the gorilla sequence being more similar than the chimpanzee. We have also found that the gorilla but not the chimpanzee contains a TYRL locus similar to the human TYRL locus. PMID- 8234204 TI - NCI cancer prevention and control research. AB - As the result of a major prevention and control effort, public health measures directed toward reducing smoking are slowly succeeding (1). Of parallel importance, and the focus of this article, is the research evidence that indicates direct and significant correlations between cancer and certain food groups such as vegetables, fruits, and fiber; the specific components in these foods that may influence cancer risk at several sites; and the intervention trials supported by the National Cancer Institute as part of its chemoprevention research program. The latter includes studies of nutrients and other food constituents as well as pharmaceutical agents that may be useful for cancer prevention. PMID- 8234205 TI - Dietary intervention and chemoprevention--1992 perspective. AB - BACKGROUND: Diet, including excess or scarcity of specific foods and nutrients, is important in the etiology of 30-60% of a variety of cancers. Dietary changes may favorably affect cancer risk and incidence. This overview summarizes recommended dietary changes and their nutritional implications and considers the problems of selecting and implementing such a program and identifying and targeting appropriate populations and subjects. RESULTS: Dietary changes that decrease the intake of fat and pickled/smoked foods, limit calorie intake and alcohol consumption, and increase the intake of fiber, fruits, and vegetables should help to decrease risk for cancers of the breast, colon, oral cavity, upper gastrointestinal tract, lung, and cervix. Micronutrients to increase include vitamins C, E, A, beta-carotene and other carotenoids, and folic acid. The diet should contain a variety of yellow-orange fruits and vegetables, green leafy and cruciferous vegetables, legumes, whole grains, low-fat dairy products, and lean meats, fish, and poultry. This diet must be adequate in proteins and minerals, palatable, affordable, and add to the quality of life. It is important that we target appropriate subjects and populations, modify foods, and design and fund research studies in areas of nutrient availability and metabolism and the biologic mechanisms of nutritional chemoprevention. CONCLUSION: A program of nutritional intervention by sound dietary changes that is effective, safe, and acceptable in chemoprevention should be the cornerstone of a cancer prevention strategy starting now. Directions for future research are discussed. PMID- 8234206 TI - Dietary supplementation: the case for placebo-controlled trials. AB - There are a variety of ways in which we learn about the relationship of diet to cancer prevention or cancer etiology in humans. Epidemiologic studies provide the basis for many of our hypotheses, while animal studies provide opportunities to try out more specific models. However, only human clinical trials can adequately test these concepts and identify which food compounds are related to cancer prevention. Further, clinical trials can provide opportunities to examine underlying mechanisms. Clinical trials in the area of diet and cancer prevention have largely been of two types: single (or dual) nutrients (purified, in high dose pills) tested in double-blind placebo-controlled trials, and randomized dietary interventions, using education of the subjects to achieve broad scale dietary change. This paper argues for an intermediate form of clinical trial, using a food-based supplement in a randomized double-blind placebo-controlled design. This model is particularly appropriate for studying food components such as fiber for which the activity may rely on higher order structure. PMID- 8234207 TI - Cancer prevention--behavior changes: the short and the long of it. AB - BACKGROUND: Behavioral factors play an important role in the adoption and maintenance of nutritional modifications, delivered either through chemopreventive regimens or through dietary adaptation. A body of research supports a protective role of fruits and vegetables, dietary fiber, and eating a low-fat diet in reducing cancer risk. There is only limited knowledge, however, about the cancer-preventing properties of specific micronutrients, apart from beta-carotene; about optimal levels of intake of differing micronutrients; and about patterns of food intake associated with reduced cancer risk. Thus, it would seem preferable, overall, to be able to recommend cancer prevention through dietary modification rather than through the administration of individual agents. METHODS: Studies of dietary adherence in cancer prevention have yielded varying success rates, but are generally quite promising. We have developed a model that encompasses the range of behavioral, psychological, social, and systemic variables thought to influence adherence to nutritional regimens. RESULTS: The model is being utilized to influence the form and content of nutritional regimens and to promote short-term change which can then be sustained as long-term lifestyle modification. The model can also be applied to evaluate adherence to nutritional or other behavioral modifications and to determine the factors predictive of success. PMID- 8234208 TI - Cancer prevention: chemoprevention vs dietary modifications. AB - BACKGROUND: The goal of cancer chemoprevention research is to identify agents which can reverse or arrest the process of carcinogenesis. Dietary supplementation with a specific agent or micro- or macronutrient has advantages over dietary manipulation. RESULTS: Prescribing a specific "pill" or supplement for long-term daily administration can lead to improved adherence over a behavior or dietary modification. Preclinical studies and clinical trials exploring pharmacokinetics, dose-response relationships, side effects, and ultimately efficacy are greatly facilitated when the agent of interest is defined. The discovery of new agents based on structure/activity relationships and mechanisms of action can be facilitated when an active agent is identified. The primary disadvantage of developing a drug as a cancer prevention agent is the potential for side effects. If active, the agent has the potential to be given to large unmonitored populations over prolonged periods of time. Any agents causing a significant increase in morbidity or mortality for whatever cause, may be more of a liability than an asset from a public health standpoint. CONCLUSION: Whether chemoprevention ultimately turns out to be a dietary constituent, a modification of a naturally occurring substance, or a synthetic compound, the recommendations for use will be based on side effect profile, dose-response, and convenience. PMID- 8234209 TI - Retinoids in chemoprevention of head and neck carcinogenesis. AB - There is currently considerable excitement about the prospects of developing effective chemoprevention strategies for head and neck cancer. The field cancerization model provides the basic assumptions which guide current head and neck chemoprevention trials. The retinoid 13-cis-retinoic acid, as well as other agents, is currently being studied in clinical trials and will hopefully establish the role of chemoprevention in head and neck cancer. PMID- 8234210 TI - Oral cancer prevention: the case for carotenoids and anti-oxidant nutrients. AB - The most convincing evidence for a preventive role for any modality is obviously demonstration of incidence reduction produced by that modality. However, cancer prevention trials with cancer incidence as an endpoint have logistic problems rendering them essentially impossible to conduct for most malignancies. Hence a workable strategy often involves analysis of other, indirect lines of evidence to reach conclusions. For oral cancer, dietary epidemiologic evidence points to a protective role for foods rich in carotenoids. Other anti-oxidants, such as vitamin C, are also implicated. Similarly, laboratory evidence points to a carcinogenesis inhibitory role for both retinoids and carotenoids. Clinical studies have targeted premalignant lesions, i.e., oral leukoplakia. For over two decades the efficacy of retinoids, natural and synthetic, has been known. Nevertheless, it has been difficult to translate this into a recommendation for prevention because of the toxicity of retinoids. The synthetic retinoid most often used in these trials is 13-cis-retinoic acid. This compound is toxic even at very low doses (0.1 mg/kg/day), particularly when given over several weeks to months. Hence, although effective, it cannot be advocated for prevention or oral cavity cancer. Studies with nontoxic antioxidants, such as beta-carotene, are much more recent. Early results are promising in that beta-carotene, alone or in combination with other nutrients, can reverse oral leukoplakia without toxicity in short-term trials. Studies currently under way will demonstrate whether durable remissions can be obtained using this strategy. It should be emphasized that such long-term trials are problematic to conduct with the toxic retinoids because the risks of prolonged exposure to them outweighs the chance of cancer development in the usual leukoplakia lesion. PMID- 8234211 TI - Highlights of the cancer chemoprevention studies in China. AB - New retinoids have been synthesized and screened in the search for chemopreventive agents of cancer. N-4-(Carboxyphenyl) retinamide showed a significant inhibitiory effect on carcinogenesis of cancers in the buccal pouch of hamsters and in the forestomach of mice. Clinical studies have demonstrated that N-4-(carboxyphenyl) retinamide is effective against oral leukoplakia, vulvar leukoplakia, and dysplasia of the uterine cervix and stomach. Field studies among a population at high risk for esophageal cancer in Linxian County, Henan Province, revealed that N-4-(ethoxycarbophenyl) retinamide decreased the incidence of this cancer. Qidong County is a high-risk area for hepatoma in China. This has been correlated to the low levels of selenium in the blood of the residents as well as in grain grown in the area. S. Y. Yu, W. G. Li, Y. J. Zhu, et al. (Biol. Trace Element Res. 1985; 7:22-26) reported that the administration of selenium inhibited the incidence of hepatoma induced by aflatoxin B in rats and in ducks. Experimental studies demonstrated that green tea extract inhibited 12-O-tetradecanoylphorbol-3-acetate-induced epidermal ornithine decarboxylase activity and counteracted 12-O-tetradecanoylphorbol-3-acetate-induced ear edema in mice. It is interesting that green tea extract inhibited the transformation of Balb/c 3T3 cells induced by methylcholanthrene and 12-O-tetradenanoylphorbol-3 acetate. Garlic has been used for thousands of years in Chinese cooking and folk medicine. Epidemiological studies show that the dietary intake of garlic is inversely related to gastric cancer incidence in Shandong Province. PMID- 8234212 TI - Retinoids or carotenoids: is there another choice? AB - Both retinoids and carotenoids decrease cancer occurrence in selected animal models of carcinogenesis, but the retinoids appear to have more potent activity against a wider variety of tumors. Further use of retinoids for cancer prevention will be limited, however, because of their toxic effects on bone and skin (among other organs). In contrast to retinoids, carotenoids seem to be free of important toxicity and this fact makes them more promising for use in the general adult population. However, the idea that carotenoids have a cancer-preventive effect in humans is based almost entirely on epidemiological studies of diet and serum which could simply reflect the effects of some other dietary constituents. In the search to explain the profoundly lower risk of death associated with eating a diet high in fruits, vegetables, and grains, there is a place for clinical trials testing one specific nutrient, such as beta-carotene, against one disease, such as lung cancer, but we should not rely exclusively on these narrowly focused studies. In addition, careful analyses of cohort studies may reveal broad patterns of diet that are associated with lower mortality. These diets can then be tested in clinical trials for their efficacy against a variety of causes of death and disability. For now, the most rational and prudent choice would be to consume a diet high in fruits, vegetables, and grains and not to take either supplemental antioxidants or retinoids until there is clear evidence of their effectiveness and safety. PMID- 8234213 TI - Controversial issues of dietary fat and experimental mammary carcinogenesis. AB - Epidemiological evidence from different countries worldwide has suggested a positive association between the availability of fat in the diet and variations in breast cancer mortality rate. A voluminous amount of information is also available in the literature linking increased fat consumption, particularly polyunsaturated fat, and stimulation of mammary tumorigenesis in animal models. In the past few years, our laboratory has been studying the impact of several confounding factors that could modulate the enhancing effect of fat on neoplastic development of the mammary gland in female rats which are treated with a carcinogen. It is our conclusion that fat promotes mammary carcinogenesis only under a very stringent set of conditions which might not be duplicated in the arena of fat intake and human breast cancer risk. Previous studies on fat and mammary cancer in experimental models have used young virgin rats which are given a dose of carcinogen at a particular age. The question arises as to whether the promoting effect of fat might be a consequence of the characteristics of the model. We have supportive evidence showing that the following criteria must be satisfied in order for fat enhancement of mammary carcinogenesis to be manifested: (a) carcinogen administered at a time when the mammary gland is exquisitely susceptible to tumor induction, (b) animals maintained on a semipurified diet, (c) ad libitum feeding necessary, and (d) unusually high requirement of linoleic acid for tumor development. On the other hand, the stimulatory effect of fat is attenuated or sometimes even negated by (a) feeding of a natural ingredient diet, (b) submaximal calorie intake, and (c) previous history of pregnancy and lactation. Given the spectrum of confounders that are inherent in epidemiological studies linking fat intake and breast cancer, including differences in lifestyle, reproductive history, eating habits, as well as complexity of the total diet, our findings suggest that there may be a need to reevaluate the validity of extrapolating animal data that are obtained under a highly defined set of conditions to the etiological significance of dietary fat in human breast cancer. PMID- 8234214 TI - Dietary fat, calories, and fiber in colon cancer. AB - The purpose of this presentation is (a) to provide an overview of the data thus far obtained in human epidemiological studies and laboratory animal models on the relationship between dietary fat, calories, and fiber and colon carcinogenesis and (b) to discuss whether the effect of dietary fat on colon carcinogenesis is due to the specific action of fat or to an associated caloric effect. Although the primary discussion of this presentation will be on laboratory animal model studies, reference will be made to human studies where appropriate. Future research will produce additional evidence for the etiologic role of types of dietary fat and fiber and total calories in cancer of the colon discussed in this presentation. PMID- 8234215 TI - Fat and calories in colon and breast cancer: from animal studies to controlled clinical trials. AB - BACKGROUND: Fat and total calories are closely correlated in the diet. Because of this correlation, studies of dietary fat and breast and colorectal cancer incidence have been peculiarly difficult to interpret. METHODS: This article reviews the information from rodent models of breast and colorectal cancer and epidemiological studies addressing the separate effects of calories and fat in the diet. Various statistical approaches described for separating the effects of calories and fat are outlined, with a review of epidemiological case-control, cohort, and prospective controlled clinical studies of diet and breast and colorectal neoplasia. RESULTS: Animal studies provide convincing evidence for a dominant effect of calorie restriction over fat modification in tumorigenesis. Individual case-control studies show no effect or are inconsistent in distinguishing calories from fat in colorectal carcinogenesis, and are largely null in breast cancer. However, a meta-analysis of these studies in breast cancer defines a relative risk (P < 0.05) of 1.40 for calories "solely" due to fat (1.48; P < 0.05). Cohort studies for colorectal cancer have reported variable results, but a recent large study of women implicates animal fats, particularly from red meat, as a risk for colon cancer, independently of calories. For breast cancer, a marginally significant effect of fat independent of calories was found in one recent cohort study, but earlier large cohort studies could not identify calories or fat as risk factors. CONCLUSION: Caloric restriction is likely to be important in inhibiting tumorigenesis based on animal studies. However, the influence of animal fats is seen both in animal studies and epidemiological studies of breast and bowel cancer. The differential effect of fats from different dietary sources in both colon and breast cancer points to a qualitative effect separate from calories in tumorigenesis. Extrapolation of significant analyses of trend to very low-fat diets should be interpreted cautiously without further information on concurrent nutrient deficiencies and competing illnesses. Randomized controlled trials of dietary intervention to prevent neoplasia are feasible and will enhance the quality of information available about diet and tumorigenesis. PMID- 8234216 TI - Colon cancer: dietary modifications required for a balanced protective diet. AB - BACKGROUND: There is increasing support of the view that our diet is too calorie dense, with its high animal fat, sugar, and alcohol content. Food processing has helped to create this situation as well as the desire to eat sugar- and fat-rich foods. By examining the influence of these dietary effects on colon cancer, experimental animal studies can help dissect the influences not readily assessable by epidemiological means. METHODS: The Sprague Dawley rat model of colon cancer induced by dimethylhydrazine provides a means of assessing dietary influences with the use of a semipurified diet and varying a single factor at a time. We have examined the influence of Ca vitamin E, protein type, and cereal dietary fiber sources on tumor burden and incidence in rats on a standardized experimental protocol. RESULTS: A significant interactive effect has been seen with high Ca and low vitamin E intake in protecting rats from tumors. When comparing differing protein sources, whey protein concentrate was found to be very protective relative to red meat and other protein sources. Spent barley grain was also shown to be very protective relative to wheat bran and commercial barley bran. CONCLUSIONS: There are several potentially useful strategies for protection from colon cancer by varying diet composition. Protein sources such as whey protein concentrate, insoluble dietary fiber from barley grain, and high calcium intake seem to be very promising. These need further detailed examination as to whether they can combine to reduce risk further and to understand better the mechanisms responsible for protection. They may provide greater potential than attempts to lower the fat in the human diet. PMID- 8234217 TI - Fat and calories in the epidemiology of colon cancer in western New York. AB - In this article, we contrast three models of macronutrients in the epidemiology of colon cancer. We then evaluate a dataset collected in Buffalo showing that the macronutrient model we use to guide our data analysis has a decisive bearing on the answers we obtain. We attempt to demonstrate that the method of statistical analysis we employ depends on the biologic-behavioral model that we assume and that, if we assume the wrong model, we may perform the wrong analysis and generate an answer that is wrong. We focus on dietary fat, because the intake of fat has received so much attention in the nutritional epidemiology of cancer. In addition, the intake of fat brings the issue of collinearity between a macronutrient and total caloric intake into focus and helps accentuate the contours of our debate. PMID- 8234218 TI - Cancer chemoprotection by oltipraz: experimental and clinical considerations. AB - Oltipraz (4-methyl-5-(2-pyrazinyl)-1,2-dithiole-3-thione) is an antischistosomal drug presently under evaluation as a possible chemoprotective agent in humans. To date, oltipraz has proved effective as an inhibitor of carcinogenesis in experimental models for breast, bladder, liver, forestomach, colon, tracheal, lung, and skin cancer. Studies on the mechanisms of action of oltipraz indicate that it affects the metabolism and disposition of chemical carcinogens, principally through the induction of electrophile detoxication enzymes. While this feature is common to many different classes of both natural and synthetic experimental chemoprotectors (i.e., phenolic antioxidants, isothiocyanates, flavonoids, indoles, cinnamates, coumarins, terpenes, and others), oltipraz may offer the earliest and easiest prospect for examining the role of enzyme induction as a protective strategy in humans. Unlike the situation with many of the anutrients, substantial preclinical research has already been conducted with oltipraz to establish its safety and efficacy in animals. Hopefully, Phase I investigations will demonstrate a high tolerance for oltipraz in the chemoprotective dose range of the drug. A major concern for the success of any trial is selecting participants who are likely to adhere to the intervention as well as to all aspects of the protocol. Factors influencing participation and adherence in a trial include the design of the trial, the nature of the disease under study as well as the nature of the intervention, in particular the toxicities of the intervention (J. A. Tangrea, M. E. Adrianza, and W. E. Helsel, Cancer Epi Biomarkers Prev 1992; 1:325-330). Chemoprotection trials with oltipraz have an excellent prospect for success. Individuals with known carcinogenic exposures are likely to be interested in participation in trials designed to reduce the risks of the exposures. Moreover, the availability of intermediate markers reflecting the modulation of the biologically effective dose of environmental carcinogens as study end points will enable efficient trials to be designed. PMID- 8234219 TI - Cancer prevention by protease inhibitors. AB - BACKGROUND: There is now compelling evidence that certain protease inhibitors have strong anticarcinogenic activity. A particular protease inhibitor derived from soybeans, the Bowman-Birk inhibitor, appears to be highly promising as a cancer chemopreventive agent. In this article, studies performed to characterize an extract of soybeans containing the Bowman-Birk inhibitor, termed Bowman-Birk inhibitor concentrate, are described. METHODS: A number of different methods have been utilized to evaluate the efficacy and safety of Bowman-Birk inhibitor concentrate as a cancer chemopreventive agent in cells and animals. Bowman-Birk inhibitor concentrate has been studied extensively in animal carcinogenesis model systems and in vitro transformation systems. RESULTS: Bowman-Birk inhibitor has been shown to prevent animal carcinogenesis in the absence of adverse side effects in several different systems. The dose proposed for human trials is approximately two orders of magnitude below doses shown to have no adverse effects in animal studies. At high doses, Bowman-Birk inhibitor concentrate has been shown to have a life-lengthening effect in mice. The intermediate marker endpoints known to be affected by Bowman-Birk inhibitor/Bowman-Birk inhibitor concentrate in in vitro and in vivo models systems include levels of oncogene expression and proteolytic activity. These intermediate marker endpoints, as well as histopathology, have been proposed for use in projected human trials which will be performed to evaluate Bowman-Birk inhibitor concentrate as an anticarcinogenic agent. CONCLUSIONS: Bowman-Birk inhibitor concentrate is now ready for human trials and is likely to emerge as a powerful chemopreventive agent for human cancer. PMID- 8234220 TI - [Microbiological synthesis of sterols]. AB - The data on preparation and application of microbial sterols are reviewed. The ways of optimization of ergosterol production are discussed. The microbiological techniques for obtaining other sterols are proposed based on using specific inhibitors and mutants. The data concerning chemical and biological transformation of yeast sterols to androstane hormones and D vitamins are presented. PMID- 8234221 TI - [Enzymatic profile of autolysing Saccharomyces yeasts]. AB - The influence of pH on the efficiency of autolysis of brewer's yeast was studied. It is shown that some kinds of peptidase activities in "solid" and "liquid" fractions of autolysates depend on the time of autolysis. The inactivation constants and activation energy of inactivation of yeast proteases were determined. It was found that heat stability of yeast proteases contained in the "solid" fraction of autolysates is significantly higher as compared to the enzyme present in the "liquid" fraction. From this results the conclusion that some of yeast proteases are immobilized on the cell wall fragments. A technique for activation of yeast enzyme systems in the course of autolysis is proposed. PMID- 8234222 TI - [Pulmonary toxicity caused by nitrofurantoin]. AB - Nitrofurantoin is used in the treatment of, and to prevent, urinary tract infections. Since this chemotherapeutic agent was introduced in the fifties, quite a number of cases of acute and chronic pulmonary damage have been reported that were conditioned by nitrofurantoin. This is a report on three cases of pulmonary nitrofurantoin reactions, one of them acute and two chronic. All three patients were examined with the inclusion of bronchoalveolar lavage and transbronchial lung biopsy. In all three cases alveolitides of varying activity were confirmed. The radiological and CT findings are described. PMID- 8234223 TI - [Criteria for determining the degree of handicap in bronchial asthma in childhood]. AB - Patients history, clinical findings, pulmonary function and necessary therapy are combined for a diagram, which should easily allow to grade childhood asthma for general practitioners, pediatricians and consultants of the health insurance. The grading system is based on the cited literature, dealing in part with the matter, and on the international consensus report on asthma. PMID- 8234224 TI - [The circadian rhythm of ciliary beat frequency of human nasal cilia in probands with healthy lungs and in patients with chronic obstructive lung disease. Includes adrenergic stimulation by terbutaline]. AB - Brush biopsies taken from the human nose eight times during 24 hours were examined under a phase contrast microscope and the ciliary beat frequency was measured directly by a photosensitive cell. Ten healthy volunteers showed in two series of measurements, at 600, 1200, 1800, 2400 initially and 300, 900, 1500, 2100 six weeks later, a marked decrease in ciliary beat frequency towards the middle of the day and a statistically higher beat frequency in the early morning. The influence of the adrenergic drug terbutaline led to a cilioexcitation ranging from 0.1-1.4 Hz (mean: 0.9 Hz). In contrast to, neither a circadian variation of the ciliary beat frequency nor a stimulation by terbutaline could be observed in a group of fifteen patients with chronic bronchitis or bronchiectasis at 600, 1200, 1800 and 2400. PMID- 8234225 TI - [Physiopathology of Respiration. 79th meeting of the working group of the German Pneumology Association. 1992. Abstracts]. PMID- 8234226 TI - Determination of the structure of symmetric coiled-coil proteins from NMR data: application of the leucine zipper proteins Jun and GCN4. AB - Previous attempts to determine the solution structures of homodimeric 'leucine zippers' using nuclear magnetic resonance (NMR) spectroscopy have been impeded by the complete symmetry of these coiled-coil molecules, which makes it impossible a priori to distinguish between intra- and intermonomer dipolar connectivities. Consequently, a number of ad hoc approaches have been used in an attempt to derive tertiary solution structures of these molecules from the NMR data. In this paper we present a more rigorous approach for analysing the NMR spectra of symmetric coiled-coil proteins. This analysis is based on calculations of intra- and intermonomer interproton distances in the recently determined crystal structure of the GCN4 leucine zipper [O'Shea, E.K., Klemm, J.D., Kim, P.S. and Alber, T. (1991) Science, 254, 539-543] and in symmetric coiled-coil models of the leucine zippers of GCN4 and the human oncoprotein Jun which we constructed using a dynamic simulated annealing approach. This analysis has enabled the formulation of a set of rules for interpreting the NMR spectra of symmetric coiled-coil proteins and has also led to the prediction of novel dipolar connectivities which we demonstrate in a 2-D NMR spectrum of the homodimeric Jun leucine zipper. PMID- 8234227 TI - An NMR study on the DNA-binding SPKK motif and a model for its interaction with DNA. AB - The solution structure of one and two repeats of the 'SPKK' DNA-binding motif is reported on the basis of NMR measurements. In dimethylsulphoxide (DMSO) the major population (approximately 90%) of peptides, SPRKSPRK(S2) and GSPKKSPRK(S2b), adopts a conformation, which has two trans prolines. The two 'SP(R/K)K' units in these peptides are equivalent and each adopts a turn structure exchanging with an extended structure. This is suggested by an NOE connectivity of the beta-turn type, between the backbone amide protons of residues (i+2) and (i+3) and NOE connectivities of the Asx(sigma)-turn type, between protons of the ith Ser and the backbone amide proton on residue (i+2). This suggests that each SP(R/K)K unit has a structural intermediate between (or a combination of) a beta-turn and an Asx(sigma)-turn. In 90-10% DMSO/H2O at 4 degrees C the two units of S2 are connected more tightly by folding into a short 3(10) helix, broken at the second proline. For another peptide, Thr-Pro-Arg-Lys(T1), the major population (75%) in 100% DMSO comprises a beta-turn in rapid exchange with an extended structure. We did not observe an NOE connectivity of the Asx(sigma) type with the T1 peptide. A possible structure of the SPKK motif in the complex with DNA is discussed. PMID- 8234228 TI - Substrate-induced activation of dienelactone hydrolase: an enzyme with a naturally occurring Cys-His-Asp triad. AB - The Cys-His-Asp catalytic triad found in dienelactone hydrolase (DLH) is unusual for several reasons. It has not been observed in other hydrolytic enzymes and it is virtually inactive when it is produced by site-directed mutagenesis in the proteases. We propose a model to explain why this triad is catalytically active in DLH but not in the proteases. In the resting state of DLH, His202 forms an ion pair with Asp171 and Cys123 exists as a thiol. The resting state thiol does not interact with His202 in the active site but instead forms a hydrogen bond with Glu36 in the interior of the molecule. In the absence of substrate, Glu36 is also ion paired with Arg206. When substrate binds, Arg206 forms a second ion pair with the anionic substrate and the Arg206/Glu36 ion pair weakens. The destabilized Glu36 carboxylate shifts towards and deprotonates the Cys123 thiol, thereby activating the nucleophile. As the thiolate anion is not energetically favoured in the hydrophobic interior of the enzyme, it swings into the active site where it can be stabilized by the His202 imidazolium and the dipole of helix C. The Cys123 thiolate which now lies adjacent to the acyl carbon of the substrate, is thus generated only in the presence of substrate. The mode of thiolate activation reduces the susceptibility of DLH towards thiol alkylating agents. PMID- 8234229 TI - The nature of the ion binding interactions in EF-hand peptide analogs: free energy simulation of Asp to Asn mutations. AB - The binding of the La3+ ion to a tridecapeptide, which is a model for the EF-hand in calcium-binding proteins, is studied in solution by free energy simulations. The calculations analyze the effect on the La3+ ion binding of the mutation of Asp to Asn for side chains that interact directly with the ion. The results are compared with the measurements of Marsden,B.J., Hodges,R.S. and Sykes,B.D. (1989) Biochemistry, 28, 8839, on the same system. They found that the Asp to Asn mutation has only a small effect on the binding; the observed differences in the free energies on changing one Asp to an Asn are between -0.3 and 1.8 kcal/mol. This result is analyzed by alchemical simulations for the tridecapeptide in the bound (loop) structure and free (extended) form. The free energy changes due to the mutation of an Asp to an Asn are large and positive for both the bound and free forms. However, since the values of the free energy changes are calculated to be similar in the two forms, the difference in the binding free energy of Asp and Asn peptides is found to be small, in agreement with experiment. By use of thermodynamic integration, the various contributions to the free energy changes are estimated. In the complexed form, the Asp to Asn mutation is favored by the reduction in the repulsive interaction with other charged residues of the peptide; it is disfavored by the reduction of the stabilization of the ion and the surrounding water has a small effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234230 TI - Protein fold refinement: building models from idealized folds using motif constraints and multiple sequence data. AB - A general solution to the problem of directly incorporating data from multiple sequence alignments into the construction of molecular models was approached through the calculation of an estimated pairwise distance based on conserved hydrophobicity. A scaling method was developed that allowed the required bulk geometric properties of the estimated pair-wise distances (mean and mean squared) to mimic those expected in a globular protein. These properties were maintained independently of the composition, length, number or degree of conservation of the original sequences. Despite being a poor estimate for individual distances were found to be compatible with the native structure and could be weighted highly. While the estimated distances provided a general drive towards hydrophobic packing, more specific structure (including secondary structures and motifs) were induced by regularization towards an ideal form. These constraints were used to refine an outline starting structure (derived only from secondary structure axes) towards a compact form that was sufficiently protein-like for side chains to be added with almost no further adjustment of the alpha-carbon positions. This process allows rough folds based on abstract representations of protein architecture to be rapidly converted to a form where they can be analysed by the growing number of methods designed to assess molecular models. PMID- 8234231 TI - Protein modelling using a chimera reference protein derived from exons. AB - Bovine pancreatic beta-trypsin (PDB ID-code: 1TPO) which is registered in the Brookhaven Protein Data Bank (PDB) consists of four exons. The results of homology searches for each exon in the PDB showed that homologous proteins were tonin (PDB ID-code: 1TON), rat mast cell protease (PDB ID-code: 3RP2__A), kallikrein A (PDB ID-code: 2PKA__B) and kallikrein A (2PKA__B) respectively. Thus, for the three-dimensional structure prediction of 1TPO, a chimera protein was constructed from the three proteins mentioned above and the 3-D structure prediction was performed using this chimera reference protein. The modelled structure of 1TPO was energetically optimized by molecular mechanics and molecular dynamics simulation and was compared with its X-ray crystal structure registered in the PDB. The root mean square deviations (r.m.s.d.) of main chain atoms and the neighbouring active site (5 A sphere from His57, Asp102 and Ser195) between the modelled structure and the X-ray structure were 1.66 and 0.94 A respectively. Porcine pancreatic elastase (PDB ID-code: 3EST) which is registered in the PDB was used as the reference protein and the modelled structure from 3EST was also compared with the X-ray data. The r.m.s.d. of main chain atoms and that of the active site were 2.14 and 1.18 A respectively. These results clearly support the propriety of this method using the chimera reference protein. PMID- 8234232 TI - Repeat of a helix-turn-helix module in DNA-binding proteins. AB - Helix-turn-helix motif is one of the common motifs observed in DNA-binding proteins. The motif interacts with DNA double helix and recognizes specific base sequences. It is assumed that the helix-turn-helix motif appears only once in seven prokaryotic transcriptional repressors of which 3-D structures have been determined by X-ray crystallographic studies. These prokaryotic repressors consist of several alpha-helices connected with turns. We report here that these repressors are decomposable into helix-turn-helix modules and their connectors. A module is defined as a compact structural unit with consecutive amino acid residues in a globular protein. Each of the helix-turn-helix motifs in the seven proteins corresponds approximately to a single helix-turn-helix module consisting of approximately 13 amino acids. Identification of modules of seven prokaryotic repressors and comparisons of their tertiary structures led to the conclusion that three of these DNA-binding proteins contain more than one helix-turn-helix module with a structure similar to the helix-turn-helix motif. The difference in module organization of these DNA-binding proteins paves the way for further classification of the DNA-binding proteins with the helix-turn-helix motif. The structural repertoire of these transcriptional regulators was increased through different utilizations in the number of helix-turn-helix and other modules. The difference in DNA base recognition ability in these helix-turn-helix modules is ascribed to a difference in size of a side chain at the fifth residue from Gly, on the turn. PMID- 8234233 TI - Predicting the point at which transmembrane helices protrude from the bilayer: a model of the antenna complexes from photosynthetic bacteria. AB - We describe a method for predicting the point at which a transmembrane helix leaves the bilayer and enters the more polar region of the aqueous exterior. This is achieved by comparing the relative directions of the hydrophobic and internal faces of the transmembrane helices which should be opposite for the regions within the bilayer but equivalent for the regions on the outside. This information provides a strong constraint in the process of modelling membrane proteins. We go on to use the approach to model the monomers of the bacterial light-harvesting antenna complexes. This information is then combined with some preliminary crystallographic data and biochemical results to produce a 3-D model of a tetramer. PMID- 8234234 TI - Pseudomonas glumae lipase: increased proteolytic stability by protein engineering. AB - The feasibility of stabilizing proteins towards proteolytic degradation was explored by engineering the primary proteolytic cleavage site(s). This novel approach does not require information on the 3-D structure of the native enzyme. As a model system, the extracellular lipase of Pseudomonas glumae was chosen, which is sensitive towards degradation by subtilisin-type proteases. The primary proteolytic cleavage in the lipase appeared to be located between amino acids serine 153 and histidine 154. Since subtilisins are known to show a preference towards amino acid residues surrounding the scissile bond, non-preferred amino acids were introduced in this area. Two concepts were tested: the introduction of arginine or glutamate residues (charge concept) and the introduction of proline residues (proline concept). Although the mutant lipases produced according to either of these concepts were still cleaved in the same area, they showed a considerably increased stability towards proteolytic degradation. PMID- 8234235 TI - Stability effects associated with the introduction of a partial and a complete Ca(2+)-binding site into human lysozyme. AB - Two mutants of human lysozyme were synthesized. Mutant A92D, in which Ala92 was substituted by Asp, contains a partial Ca(2+)-binding site and mutant M4, in which Ala83, Gln86, Asn88 and Ala92 were replaced by Lys, Asp, Asp and Asp respectively, contains the complete Ca(2+)-binding site of bovine alpha lactalbumin. The Ca(2+)-binding constants of wild type human lysozyme and of mutants A92D and M4, measured at 25 degrees C and pH 7.5, were 2(+/- 1) x 10(2) M 1, 8(+/- 2) x 10(3) M-1 and 9(+/- 0.5) x 10(6) M-1 respectively. Information gathered from microcalorimetric and CD spectroscopic measurements indicates that the conformational changes of the M4 mutant lysozyme, induced by Ca2+ binding, are smaller than those observed for bovine alpha-lactalbumin and for the Ca(2+) binding equine lysozyme. At pH 4.5, the thermostability of both the apo and Ca2+ forms of the A92D human was decreased in comparison with that of native human lysozyme. In particular, within the apo form of this mutant an alpha-helix containing sequence was destabilized. In contrast, at the same pH the thermostability of the apo and Ca2+ forms of the M4 mutant lysozyme was increased. The epsilon-ammonium group of the Lys83 side chain is assumed to be responsible for the stabilization of the apo form of this mutant. PMID- 8234236 TI - The role of asparagine-32 in forming the receptor-binding epitope of human epidermal growth factor. AB - The highly conserved asparagine residue at position 32 (Asn32) in the 'hinge' region of epidermal growth factor (EGF) separates the N- and C-terminal structural motifs of the EGF molecule and is therefore an appropriate target for structure-function studies. Analogs of human EGF (hEGF) were generated in which Asn32 was substituted with aspartate, glycine, isoleucine, lysine, proline and tryptophan. The relative affinity of the EGF receptor for mutant hEGF analogs was determined by radioreceptor competition assay. A wide range of receptor affinities was observed depending on the amino acid substitution. N32K and N32W hEGF analogs had relatively high receptor affinity, while the N32G and N32D analogs showed decreased affinity, 35% and 25% respectively, relative to wild type hEGF. However, no binding of the N32P analog was detected by radioreceptor competition assay. The N32P mutant displayed an NMR spectrum significantly different from that of native wild type hEGF, indicating gross structural perturbation. In contrast, the N32K and N32D analogs exhibited spectra similar to that of native wild type hEGF. Genetically combining the N32D hEGF with an hEGF species having either the mutation L26G in the N-terminal region or L47A in the C terminal region, generated double-mutant hEGF species which had relative affinities essentially equal to the product of the relative affinities of the parent hEGF mutants, indicating functionally independent changes in ligand receptor interaction. These studies indicate the requirement for H-bond donor functionality in the side chain of residue number 32 in forming a fully competent receptor-binding epitope. PMID- 8234237 TI - Efficient production of recombinant human factor VIII by co-expression of the heavy and light chains. AB - We have developed a high-level expression system for human blood coagulation factor VIII (FVIII) consisting of a 90 kDa heavy (H-)chain and an 80 kDa light (L )chain. Two expression plasmids were prepared, one expressing the H-chain and the other expressing the L-chain. These recombinant plasmids were designed to produce each chain linked to short additional amino acid residues derived from the FVIII precursor sequence. Furthermore, Kozak's translation initiation consensus sequence was introduced into the start codon for the H-chain. These modifications have dramatically increased the levels of expression of these chains. Chinese hamster ovary (CHO) cells co-transfected with these two recombinant plasmids were subjected to gene amplification and cloning. The final cell line, designated CTC CF8, secretes 15 IU/day/10(6) cells of active FVIII which is indistinguishable from plasma-derived FVIII in its structure and biochemical properties. This system is suitable for large-scale production of pathogen-free recombinant human FVIII which can be used for the treatment of haemophilia A patients. PMID- 8234238 TI - The limits of simulations. PMID- 8234239 TI - Automated docking in crystallography: analysis of the substrates of aconitase. AB - Automated docking of substrates to proteins of known structure aids the process of crystallographic analysis in two ways. First, automated docking can be used to generate a small number of starting models for substrates using only protein coordinates from an early stage of refinement. Second, automated docking provides a method for exploring aspects of catalysis that are inaccessible to crystallography by postulating binding modes of catalytic intermediates. This paper describes the use of automated docking to explore the binding of substrates to aconitase. The technique starts with a substrate molecule in an arbitrary configuration and position and finds favorable docked configurations in a (static) protein active site based on a molecular mechanics type force field. Using protein coordinates from an early stage of refinement of an aconitase isocitrate complex, we successfully predicted the binding configuration of isocitrate. Four configurations were found, the energetically most favorable of which fit the observed electron density well and was used as a starting model for further refinement. Two configurations were found in citrate docking experiments, the second of which approximates the mode of substrate binding in an aconitase nitrocitrate complex. We were also able to propose two binding modes of the catalytic intermediate cis-aconitate. These correspond closely to the isocitrate and the citrate binding modes. The relation of these new results to the proposed reaction mechanism is discussed. PMID- 8234240 TI - Crystallization and preliminary X-ray crystallographic analysis of chitinase from barley seeds. AB - Chitinase from barley seeds has been crystallized at room temperature using polyethylene glycol as precipitant. The crystal is monoclinic, belonging to the space group P2(1), with unit cell parameters of a = 69.43 A, b = 44.55 A, c = 81.41 A, and beta = 111.95 degrees. The asymmetric unit seems to contain two molecules of chitinase with a corresponding crystal volume per protein mass (VM) of 2.25 A3/Da and a solvent content of 45% by volume. The crystal diffracts to at least 2.0 A with X-rays from a rotating anode source and is very stable in the X ray beam. X-ray data have been collected to better than 2.2 A Bragg spacing from a native crystal. PMID- 8234241 TI - A computer model of the interleukin-4/receptor complex. AB - Interleukin-4 is a member of the cytokine family, a group of related messenger proteins which collectively help to moderate and control the immune response. It is believed that the folding topology of the beta-sheets of the interleukin-4 receptor (IL4R) is the same as that seen in the crystal structure of CD4. Although the sequence identity is low, homology modeling techniques have been used to model the IL4R structure from CD4. Refinement by molecular dynamics leads to a suggested structure which has been docked to interleukin-4 (IL4). Several residues of apparent importance for binding are identified. PMID- 8234242 TI - NMR docking of the competitive inhibitor thymidine 3',5'-diphosphate into the X ray structure of staphylococcal nuclease. AB - In the X-ray structure of the ternary staphylococcal nuclease-Ca(2+)-3',5'-pdTp complex, the conformation of the bound inhibitor 3',5'-pdTp is distorted by Lys 70* and Lys-71* from an adjacent molecule of the enzyme in the crystal lattice (Loll, P. J. and Lattman, E. E. Proteins 5:183-201, 1989; Serpersu, E. H., Hibler, D. W., Gerlt, J. A., and Mildvan, A. S. Biochemistry 28:1539-1548, 1989). Since this interaction does not occur in solution, the NMR docking procedure has been used to correct this problem. Based on 8 Co(2+)-nucleus distances measured by paramagnetic effects on T1, and 9 measured and 45 lower limit interproton distances determined by 1D and 2D NOE studies of the ternary Ca2+ complex, the conformation of enzyme-bound 3',5'-pdTp is high-anti (chi = 58 +/- 10 degrees) with a C2' endo/O1' endo sugar pucker (delta = 143 +/- 2 degrees), (-) synclinal about the C3'-O3' bond (epsilon = 273 +/- 4 degrees), trans, gauche about the C4' C5' bond (gamma = 301 +/- 29 degrees) and either (-) or (+) clinal about the C5' O5' bond (beta = 92 +/- 8 degrees or 274 +/- 3 degrees). The structure of 3',5' pdTp in the crystalline complex differs due to rotations about the C4'-C5' bond (gamma = 186 +/- 12 degrees, gauche, trans) and the C5'-O5' bond [beta = 136 +/- 10 degrees, (+) anticlinal]. The undistorted conformation of enzyme-bound metal 3',5'-pdTp determined by NMR was docked into the X-ray structure of the enzyme, using 19 intermolecular NOEs from ring proton resonances of Tyr-85, Tyr-113, and Tyr-115 to proton resonances of the inhibitor. van der Waals overlaps were then removed by energy minimization. Subsequent molecular dynamics and energy minimization produced no significant changes, indicating the structure to be in a global rather than in a local minimum. While the metal-coordinated 5'-phosphate of the NMR-docked structure of 3',5'-pdTp overlaps with that in the X-ray structure, and similarly receives bifunctional hydrogen bonds from both Arg-35 and Arg-87, the thymine, deoxyribose, and 3'-phosphate are significantly displaced from their positions in the X-ray structure, with the 3'-phosphate receiving hydrogen bonds from Lys-49 rather than from Lys-84 and Tyr-85. The repositioned thymine ring permits hydrogen bonding to the phenolic hydroxyl of Tyr-115.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234243 TI - Mutational tests of the NMR-docked structure of the staphylococcal nuclease-metal 3',5'-pdTp complex. AB - In the X-ray structure of the staphylococcal nuclease-Ca(2+)-3',5'-pdTp complex, the conformation of the inhibitor 3',5'-pdTp is distorted by Lys-70* and Lys-71* from an adjacent molecule of staphylococcal nuclease (Loll, P.J., Lattman, E.E. Proteins 5:183-201, 1989). In order to correct this crystal packing problem, the solution conformation of enzyme-bound 3',5'-pdTp in the staphylococcal nuclease metal-pdTp complex determined by NMR methods was docked into the X-ray structure of the enzyme [Weber, D.J., Serpersu, E.H., Gittis, A.G., Lattman, E.E., Mildvan, A.S. (preceding paper)]. In the NMR-docked structure, the 5'-phosphate of 3',5' pdTp overlaps with that in the X-ray structure. However, the 3'-phosphate accepts a hydrogen bond from Lys-49 (2.89 A) rather than from Lys-84 (8.63 A), and N3 of thymine donates a hydrogen bond to the OH of Tyr-115 (3.16 A) which does not occur in the X-ray structure (5.28 A). These interactions have been tested by binding studies of 3',5'-pdTp, Ca2+, and Mn2+ to the K49A, K84A, and Y115A mutants of staphylococcal nuclease using water proton relaxation rate and EPR methods. Each mutant was fully active and structurally intact, as found by CD and two-dimensional NMR spectroscopy, but bound Ca2+ 9.1- to 9.9-fold more weakly than the wild-type enzyme. While the K84A mutation did not significantly weaken 3',5'-pdTp binding to the enzyme (1.5 +/- 0.7 fold), the K49A mutation weakened 3',5'-pdTp binding to the enzyme by the factor of 4.4 +/- 1.8-fold. Similarly, the Y115A mutation weakened 3',5'-pdTp binding to the enzyme 3.6 +/- 1.6-fold. Comparable weakening effects of these mutations were found on the binding of Ca(2+)-3',5'-pdTp. These results are more readily explained by the NMR-docked structure of staphylococcal nuclease-metal-3',5'-pdTp than by the X-ray structure. PMID- 8234244 TI - Performance evaluation of amino acid substitution matrices. AB - Several choices of amino acid substitution matrices are currently available for searching and alignment applications. These choices were evaluated using the BLAST searching program, which is extremely sensitive to differences among matrices, and the Prosite catalog, which lists members of hundreds of protein families. Matrices derived directly from either sequence-based or structure-based alignments of distantly related proteins performed much better overall than extrapolated matrices based on the Dayhoff evolutionary model. Similar results were obtained with the FASTA searching program. Improved performance appears to be general rather than family-specific, reflecting improved accuracy in scoring alignments. An implementation of a multiple matrix strategy was also tested. While no combination of three matrices performed as well as the single best matrix, BLOSUM 62, good results were obtained using a combination of sequence based and structure-based matrices. This hybrid set of matrices is likely to be useful in certain situations. Our results illustrate the importance of matrix selection and the value of a comprehensive approach to evaluation of protein comparison tools. PMID- 8234245 TI - A homology model of human interferon alpha-2. AB - An atomic coordinate five alpha-helix three-dimensional model is presented for human interferon alpha-2 (HuIFN alpha 2). The HuIFN alpha 2 structure was constructed from murine interferon beta (MuIFN beta) by homology modeling using the STEREO and IMPACT programs. The HuIFN alpha 2 model is consistent with its known biochemical and biophysical properties including epitope mapping. Lysine residues predicted to be buried in the model were primarily unreactive with succinimidyl-7-amino-4-methylcoumarin-3-acetic acid (AMCA-NHS), a lysine modification agent, as shown by mass spectrometric analysis of tryptic digests. N terminal sequence analysis of polypeptides generated by limited digestion of HuIFN alpha 2 with endoproteinase Lys-C demonstrated rapid cleavage at K31, which is consistent with the presence of this residue in a loop in the proposed HuIFN alpha 2 model. Based on this model structure potential receptor binding sites are identified. PMID- 8234246 TI - Primary structure effects on peptide group hydrogen exchange. AB - The rate of exchange of peptide group NH hydrogens with the hydrogens of aqueous solvent is sensitive to neighboring side chains. To evaluate the effects of protein side chains, all 20 naturally occurring amino acids were studied using dipeptide models. Both inductive and steric blocking effects are apparent. The additivity of nearest-neighbor blocking and inductive effects was tested in oligo and polypeptides and, surprisingly, confirmed. Reference rates for alanine containing peptides were determined and effects of temperature considered. These results provide the information necessary to evaluate measured protein NH to ND exchange rates by comparing them with rates to be expected for the same amino acid sequence is unstructured oligo- and polypeptides. The application of this approach to protein studies is discussed. PMID- 8234247 TI - Isotope effects in peptide group hydrogen exchange. AB - Kinetic and equilibrium isotope effects in peptide group hydrogen exchange reactions were evaluated. Unlike many other reactions, kinetic isotope effects in amide hydrogen exchange are small because exchange pathways are not limited by bond-breaking steps. Rate constants for the acid-catalyzed exchange of peptide group NH, ND, and NT in H2O are essentially identical, but a solvent isotope effect doubles the rate in D2O. Rate constants for base-catalyzed exchange in H2O decrease slowly in the order NH > ND > NT. The alkaline rate constant in D2O is very close to that in H2O when account is taken of the glass electrode pH artifact and the difference in solvent ionization constant. Small equilibrium isotope effects lead to an excess equilibrium accumulation of the heavier isotopes by the peptide group. Results obtained are expressed in terms of rate constants for the random coil polypeptide, poly-DL-alanine, to provide reference rates for protein hydrogen exchange studies as described in Bai et al. [preceding paper in this issued]. PMID- 8234248 TI - Daidzin and daidzein suppress free-choice ethanol intake by Syrian golden hamsters. AB - Syrian Golden hamsters prefer and consume large and remarkably constant amounts of ethanol in a simple two-bottle free-choice regimen. Ethanol intake is significantly suppressed by zimelidine, bromocriptine, buspirone, and lithium carbonate, pharmacological agents that have been shown to be beneficial in controlling ethanol intake in alcohol-dependent humans. These results suggest that this ethanol-drinking animal model has high "predictive validity" and can be used effectively in the search for and identification of new agents for the treatment of alcohol abuse. The model has enabled us to confirm the putative antidipsotropic effect of Radix puerariae (RP), an herb long used in traditional Chinese medicine for the treatment of patients who abuse alcohol. A crude extract of RP at a dose of 1.5 g.kg-1 x day-1 significantly suppresses (> 50%) the free choice ethanol intake of Golden hamsters. Moreover, two major constituents of RP, daidzein (4',7-dihydroxyisoflavone) and daidzin (the 7-glucoside of daidzein), were also shown to suppress free-choice ethanol intake. Daidzin and daidzein, at doses of 150 and 230 mg.kg-1 x day-1, respectively, suppress ethanol intake by > 50%. RP, daidzein, and daidzin treatment do not significantly affect the body weight and water or food intake of the hamsters. These findings identify a class of compounds that offer promise as safe and effective therapeutic agents for alcohol abuse. PMID- 8234249 TI - Oligonucleotide clamps arrest DNA synthesis on a single-stranded DNA target. AB - Triple helices can be formed on single-stranded oligopurine target sequences by composite oligonucleotides consisting of two oligonucleotides covalently linked by either a hexaethylene glycol linker or an oligonucleotide sequence. The first oligomer forms Watson-Crick base pairs with the target, while the second oligomer engages in Hoogsteen base pairing, thereby acting as a molecular clamp. The triple-helical complex formed by such an oligonucleotide clamp, or "oligonucleotide-loop-oligonucleotide" (OLO), is more stable than either the corresponding trimolecular triple helix or the double helix formed upon binding of the oligopyrimidine complement to the same oligopurine target. Attaching a psoralen derivative to the 5' end of the OLO allowed us to photoinduce a covalent linkage to the target sequence. The psoralen moiety became covalently linked to all three portions of the triplex, thereby making the oligonucleotide clamp irreversible. These crosslinking reactions introduced strong stop signals during DNA replication, as shown on a plasmid containing a portion of the HIV proviral sequence of human immunodeficiency virus. A 16-mer oligopurine sequence corresponding to the "polypurine tract" of human immunodeficiency virus was chosen as a target for a psoralen-OLO conjugate. Three different stop signals for DNA polymerase were observed, corresponding to different sites of polymerase arrest on its template. Even in the absence of photoinduced crosslinking, the psoralen-OLO conjugate was able to arrest DNA replication. The formation of triple-helical structures on single-stranded targets may provide an alternative to the antisense strategy for the control of gene expression. PMID- 8234250 TI - An unusual active hexose transport system in human and mouse preimplantation embryos. AB - In a metabolic study of human and mouse preimplantation embryos (preembryos), we measured glucose uptake and phosphorylation with nonradioactive 2-deoxyglucose (DG) as tracer. Initial experiments indicated an active hexose transport capacity, a property thought to be restricted in mammals to intestinal villi and kidney tubules [Baly, D. L. & Horuk, R. (1988) Biochim. Biophys. Acta 947, 571 590]. Significant findings are as follows: (i) During a 60-min incubation with a low level of DG, mouse blastocyst DG rose to levels up to 30 times that of the medium. (The intestinal active system does not transport DG [Crane, R. K. (1960) Physiol. Rev. 40, 789-825].) (ii) Active preembryo transport was not blocked (as it would have been in the intestine) by phlorizin [Alvarado, F. & Crane, R. K. (1962) Biochem. Biophys. Acta 56, 170-172 and Sacktor, B. (1989) Kidney Int. 36, 342-350] or by replacement of Na+ with choline+ or K+ [Crane (1960) and Sacktor (1989)]. (iii) Transport of DG was blocked by cytochalasin B (which is not true for the intestinal transporter). We conclude that a distinct active hexose transporter and at least one facilitated transporter are present in preembryos, perhaps appearing in tandem on different membranes during formation of the increasingly complex preembryo structure. PMID- 8234251 TI - Amino acid sequence of a putative sodium channel expressed in the giant axon of the squid Loligo opalescens. AB - A full-length cDNA encoding a putative Na+ channel (GFLN1) has been cloned from a library prepared from the stellate ganglion of Loligo opalescens. The cDNA encodes a predicted protein of 1784 amino acids. Regions of the GFLN1 protein with defined functional importance (membrane span S4, the SS1 and SS2 segments, and interdomain III-IV) are highly conserved among all vertebrate Na+ channel alpha-subunit structures. Northern blot hybridization and RNase protection assays verify that mRNA corresponding to GFLN1 is expressed in neurons of the giant fiber lobe that form the giant axon. We propose that GFLN1 encodes the Na+ channel that has been extensively studied in the squid axon. PMID- 8234252 TI - Linking yeast genetics to mammalian genomes: identification and mapping of the human homolog of CDC27 via the expressed sequence tag (EST) data base. AB - We describe a strategy for quickly identifying and positionally mapping human homologs of yeast genes to cross-reference the biological and genetic information known about yeast genes to mammalian chromosomal maps. Optimized computer search methods have been developed to scan the rapidly expanding expressed sequence tag (EST) data base to find human open reading frames related to yeast protein sequence queries. These methods take advantage of the newly developed BLOSUM scoring matrices and the query masking function SEG. The corresponding human cDNA is then used to obtain a high-resolution map position on human and mouse chromosomes, providing the links between yeast genetic analysis and mapped mammalian loci. By using these methods, a human homolog of Saccharomyces cerevisiae CDC27 has been identified and mapped to human chromosome 17 and mouse chromosome 11 between the Pkca and Erbb-2 genes. Human CDC27 encodes an 823-aa protein with global similarity to its fungal homologs CDC27, nuc2+, and BimA. Comprehensive cross-referencing of genes and mutant phenotypes described in humans, mice, and yeast should accelerate the study of normal eukaryotic biology and human disease states. PMID- 8234253 TI - Heterologous expression of specific K+ channels in T lymphocytes: functional consequences for volume regulation. AB - It has been postulated that the K+ channel isoform Kv1.3 plays a role in regulatory volume decrease (RVD) in response to hypotonic shock. We show that a mouse cytotoxic T-lymphocyte line, CTLL-2, is devoid of voltage-dependent K+ channels and is unable to volume regulate. Transient transfection of these cells with Kv1.3 reconstitutes their ability to volume regulate. As predicted by our model, this ability depends critically on volume-induced changes in membrane potential and the isoform of the K+ channel used. When the cells were transfected with Kv3.1, an isoform believed to be expressed in a specific subclass of mouse thymocytes, the CTLL-2 cells did not show RVD. The difference in the ability of the two isoforms to confer the capacity for RVD is expected from differences in the voltage dependence of activation of the channels, according to our proposed model for RVD. The experimental approach that we use, transient transfection and panning to select positive transfectants, is highly effective; it has a > 95% efficiency. This method, and this cell line, may be important tools in studying lymphocyte K+ channels and their function in situ. PMID- 8234254 TI - Selective inhibition of the carotid body sensory response to hypoxia by the substance P receptor antagonist CP-96,345. AB - Carotid bodies are sensory organs for monitoring arterial oxygen and CO2. Previous studies have shown that chemoreceptor tissue contains substance P (SP) and exogenously administered SP augments chemosensory discharge. In the present study, we examined the physiological importance of SP in carotid body chemoreception by using a selective nonpeptide SP [neurokinin (NK) 1] receptor antagonist CP-96,345. In experiments performed on anesthetized cats, sensory discharge was recorded from the carotid body in situ. To control for alterations in blood flow, additional studies were conducted on the carotid body in vitro. In in vivo studies, close carotid body (intraarterial) administration of CP-96,345 attenuated the sensory response to hypoxia in a dose-dependent manner with 73% of the response abolished at doses of 0.3-0.6 mg/kg. Comparable doses of the (2R,3R) enantiomer had no effect on hypoxia-induced excitation, indicating that the effect of CP-96,345 was not due to nonspecific action. In contrast, the carotid body response to high CO2 was not affected by CP-96,345, implying that only the hypoxic response is mediated by NK-1 receptor and confirming that the effect of the SP antagonist was not due to nonspecific actions. Marked attenuation of the sensory response to hypoxia was also obtained in the carotid body in vitro, suggesting that the effects of the NK-1 antagonist were not secondary to cardiovascular changes. These results demonstrate that CP-96,345 attenuates or abolishes the chemosensory response to hypoxia but not to CO2 and suggest that SP mediates the hypoxia-induced sensory excitation in the cat carotid body via NK-1 receptor activation. PMID- 8234255 TI - The role of complement component C3b and its receptors in sperm-oocyte interaction. AB - Previous studies have shown that human sperm that have undergone the acrosome reaction express a unique tissue-specific variant of the complement component 3 (C3)-binding molecule membrane cofactor protein (MCP, CD46) and that damaged or dead sperm activate the alternative pathway of complement and bind C3 catabolites. In this study we provide evidence that MCP on sperm that have undergone the acrosome reaction specifically binds dimeric C3b and that human sperm acrosomal proteases released during the acrosome reaction directly cleave C3, facilitating its binding to MCP. Furthermore, human and hamster oocytes can activate the alternative pathway of complement and also bind human C3 fragments. Monoclonal antibodies specific for complement receptors type 1 (CD35) and type 3 (CD11b/CD18) bind to the human oocyte plasma membrane, indicating that specific complement-binding molecules may play a role in the attachment of C3 catabolites to oocytes. Subsaturating concentrations of dimeric C3b (0.01-1 microM) promoted penetration of hamster oocytes by human sperm, whereas saturating doses (> 10 microM) inhibited this process. In addition, antibodies to both MCP and C3 significantly inhibited penetration of hamster oocytes by human sperm. These data provide evidence that regulated gamete-induced generation of C3 fragments and the binding of these fragments by selectively expressed receptors on sperm and oocytes may be an initial step in gamete interaction, leading to membrane fusion and fertilization. PMID- 8234256 TI - Molecular cloning and functional expression of a cDNA encoding glycosylation inhibiting factor. AB - By using probes based on partial amino acid sequence of glycosylation-inhibiting factor (GIF) from a mouse T-cell hybridoma, a full-length cDNA encoding mouse GIF was isolated. A cDNA clone encoding human GIF was isolated from cDNA libraries of a GIF-producing human T-cell hybridoma by using mouse GIF cDNA as a probe. The cDNAs encode a putative 12.5-kDa peptide of 115 amino acids. Northern blot analysis demonstrated a single, 0.6-kb transcript. Polyclonal rabbit antibodies against the Escherichia coli-derived recombinant 13-kDa peptide bound hybridoma derived GIF. Although the peptide did not contain a signal peptide sequence, transfection of the cDNA into COS-1 cells resulted in secretion of 13-kDa peptide, but the peptide had substantially less bioactivity than the hybridoma derived GIF. However, expression of a chimeric cDNA encoding a fusion protein consisting of the N-terminal pro region of calcitonin precursor and human GIF and cotransfection with furin cDNA to allow intracellular cleavage of the fusion protein resulted in secretion of 13-kDa peptide that was comparable to hybridoma derived GIF in its bioactivity. Both the 13-kDa peptide and GIF bioactivity in the transfected COS-1 supernatant bound to a monoclonal antibody against hybridoma-derived human GIF. These results indicate that the 13-kDa peptide represents recombinant GIF, but posttranslational modification of the peptide is important for generation of the bioactivity. The GIF cDNA had high homology with the cDNA encoding macrophage migration inhibitory factor. However, the recombinant GIF failed to inhibit migration of human monocytes, and recombinant human macrophage migration inhibitory factor did not have GIF bioactivity. PMID- 8234257 TI - Characterization of a gene encoding a Ca(2+)-ATPase-like protein in the plastid envelope. AB - By screening an Arabidopsis expression library with an antiserum against chloroplast envelope proteins, we have isolated a partial cDNA with an open reading frame that encodes a polypeptide similar to P-type cation-transporting ATPases. The corresponding genomic clone was isolated and the complete coding sequence was deduced after identification and mapping of introns. The gene has been designated PEA1 (plastid envelope ATPase) and the predicted polypeptide PEA1p. PEA1p has 946 amino acids and a molecular mass of 104 kDa. This protein is 40-44% identical to various mammalian plasma membrane Ca(2+)-ATPases but lacks the C-terminal calmodulin binding domain present in the mammalian polypeptides. When aligned with mammalian plasma membrane Ca(2+)-ATPases, PEA1p has a 70- to 80 amino acid N-terminal region that extends beyond the N terminus of these enzymes. This extension has some similarity to the transit peptide of the plastid envelope phosphate translocator and may function to target the protein to the plastid. Antibodies raised against a portion of PEA1p recognize a single 90- to 95-kDa polypeptide in chloroplast inner envelope preparations. Transcript abundance as determined by RNase protection was found to be 7- to 9-fold higher in roots than in leaves. Possible roles for a plastid envelope calcium pump are suggested. PMID- 8234258 TI - Molecular cloning of the human and monkey sperm surface protein PH-20. AB - The guinea pig sperm surface protein PH-20 has an essential function in sperm adhesion to the zona pellucida of guinea pig eggs. Fully effective contraception has been achieved by immunizing either male or female guinea pigs with purified guinea pig PH-20. Here we report the isolation of human and cynomolgus monkey PH 20 cDNAs as a key step toward testing the function of primate PH-20 and the contraceptive efficacy of PH-20 immunization in primates. The deduced amino acid sequence of human PH-20 has 509 residues and is 59% identical with guinea pig PH 20, suggesting they may have a conserved function and immunogenicity. Southern blots show that there is a single PH-20 gene in the human genome and Northern blots of human testis poly(A)+ RNA show a 2.4-kb message. Northern blots of tissues other than testis are negative for PH-20, indicating that human PH-20 is testis-specific. PMID- 8234259 TI - Ets oncogene-related gene Elg functions in Drosophila oogenesis. AB - Members of the ets gene family encode transcription factors that regulate the expression of a variety of cellular and viral genes including several protooncogenes. We have utilized Drosophila to elucidate the in vivo function of one family member. We show by complementation rescue and sequence analysis that the female sterile mutant tiny eggs (tne) is an allele of the Drosophila Ets related gene Elg (also called D-elg). The mutation of a highly conserved tyrosine residue in the ETS DNA-binding domain of the Elg gene product demonstrates that normal gene function is required for proper follicle cell migration, chorion formation, and nurse cell-chromosome decondensation during Drosophila oogenesis. PMID- 8234260 TI - Phospholipase D-catalyzed hydrolysis of phosphatidylcholine provides the choline precursor for acetylcholine synthesis in a human neuronal cell line. AB - To identify the metabolic pathway that generates choline (Cho) for acetylcholine (AcCho) from its storage pool in membrane phosphatidylcholine (PtdCho), human neuronal cells (LA-N-2) were radioisotopically labeled with 1-O-hexadecyl-2 hydroxy-sn-glycero(3)phospho[14C]choline. The compound was efficiently taken up by the cells and metabolically labelled PtdCho, Cho, AcCho, and phosphocholine pools. In pulse-chase experiments, the specific radioactivities of the metabolites of 1-O-hexadecyl-2-hydroxy-sn-glycero(3)-phospho[14C]choline indicated that it was rapidly acylated to Ptd-Cho and then hydrolyzed first to free Cho and not to phosphocholine or glycerophosphocholine. This Cho was subsequently converted to AcCho and to phosphocholine. In the absence of exogenous Cho, at least 15% of the total cellular AcCho pool was synthesized by this pathway in 1 h. The data demonstrate that the liberation of the free Cho precursor for AcCho synthesis from PtdCho can be accomplished in a one-step process, indicating the involvement of a phospholipase D-type enzyme. In the presence of hemicholinium-3, which inhibits Cho transport, the amount of intracellular [14C]Cho metabolites that accumulated during the chase period was higher than in control cells, indicating that PtdCho hydrolysis liberated Cho directly into the cytoplasm. These data show that cholinergic cells are characterized by an intracellular pathway, catalyzed by a phospholipase D, that generates Cho for AcCho synthesis from PtdCho. Abnormalities in the regulation of this pathway may contribute to selective vulnerability of cholinergic neurons in certain neurodegenerative diseases, e.g., Alzheimer disease. PMID- 8234261 TI - Determination of mRNA fate by different RNA polymerase II promoters. AB - Translational stop mutations of the human beta-globin gene cause a reduction of cytoplasmic mRNA accumulation in thalassemia patients and in transfection models. The exact mechanism underlying this phenomenon has remained enigmatic but is known to be post-transcriptional. We have used transfected HeLa cells to study the expression of beta-globin mRNAs with nonsense or frameshift mutations within the three exons of this gene. Mutations in exons 1 or 2 reduce cytoplasmic mRNA accumulation whereas a mutation in exon 3 permits essentially normal expression. We report here that the post-transcriptional fate of mutated beta-globin mRNAs is differentially affected by the type of RNA polymerase II promoter driving expression. Replacement of the beta-globin promoter with the herpes simplex virus type 1 thymidine kinase gene promoter but not the cytomegalovirus immediate early promoter rescues the cytoplasmic accumulation of mutated mRNA to wild-type levels. This effect is shown to be independent of the absolute quantity and the kinetics of accumulation of mutated mRNA synthesized, and primer-extension analyses confirm that both viral promoters accurately utilize identical transcription start sites. These data thus reveal an unexpected property of RNA polymerase II promoters: determination of the post-transcriptional fate of the maturing mRNA, presumably by influencing alternative choices between as yet undefined processing and/or transport pathways. PMID- 8234262 TI - Disruption of mitotic spindle orientation in a yeast dynein mutant. AB - Dynein motor isoforms have been implicated as potential kinetochore-associated motors that power chromosome-to-pole movements during mitosis. The recent identification and sequence determination of genes encoding dynein isoforms has now permitted the in vivo analysis of dynein function in mitosis. In this report we describe the identification and mutational analysis of the gene, DHC1, encoding a dynein heavy chain isoform in Saccharomyces cerevisiae. Sequence analysis of a 9-kb genomic fragment of the DHC1 gene predicts a polypeptide highly homologous to dynein sequences characterized from sea urchin, Dictyostelium, Drosophila, and rat. Mutations in the yeast dynein gene disrupt the normal movement of the spindle into budding daughter cells but have no apparent effect on spindle assembly, spindle elongation, or chromosome segregation. Our results suggest that, in yeast, a dynein microtubule motor protein has a nonessential role in spindle assembly and chromosome movement but is involved in establishing the proper spindle orientation during cell division. PMID- 8234263 TI - Sequencing using pulsed field and image reconstruction. AB - The use of pulsed fields in a standard manual sequencing set-up results in the separation of > 2 kb on a single gel, as compared to 300-400 bases with a dc field. However, visual reading of the sequence from a film exposed to a pulsed field gel is not possible for more than 800-900 bases under the best conditions. The use of image reconstruction and enhancement techniques allows the reading of the M13mp18 sequence to > 1 kb, and individual bands can be identified at > 2 kb. PMID- 8234264 TI - A 4.5-megabase yeast artificial chromosome contig from human chromosome 13q14.3 ordering 9 polymorphic microsatellites (22 sequence-tagged sites) tightly linked to the Wilson disease locus. AB - We have previously performed a genetic analysis of multiply affected families to map a locus responsible for Wilson disease (WND) to a 0.3-centimorgan (cM) region within chromosome 13q14.3, between D13S31 and D13S59. Here we describe the construction of a contig of approximately 4.5 Mb, which spans this region and extends from D13S25 to D13S59. This contig consists of 28 genomic yeast artificial chromosome (YAC) clones. Five critical crossover events have been defined in this interval in two unaffected (Centre d'Etudes du Polymorphisme Humain) and three WND families. The combination of sequence tagged site content mapping of YACs with both polymorphic and nonpolymorphic markers and recombination breakpoint mapping resulted in the following order of polymorphic markers: centromere-RB1-D13S25-AFM205vh2-D13S31-D13S22 7-D13S228-AFM238vc3 D13S133- AFM084xc5-D13S137-D13S169, D13S155-D13S59-telomere. The recombination/physical distance ratio varies from approximately 3000 kb per cM in the region between D13S31 and D13S25 to 6000 kb per cM in the region between D13S31 and D13S59. Three WND families exhibiting recombination between the disease locus and D13S31 or D13S59 were genotyped for additional markers in this region and further refined the location of the WND gene to between D13S155 and D13S133. Nine of the markers in this region of < 1 cM are polymorphic microsatellites (seven have observed heterozygosities of 70% or above) that will be extremely useful in prenatal and preclinical diagnosis of this disease. This physical map is an essential step in the isolation of the WND gene and is a framework for the identification of candidate genes. PMID- 8234265 TI - Identification and characterization of hematopoietic stem cells from the yolk sac of the early mouse embryo. AB - The yolk sac is the first site of hematopoiesis in the mammalian embryo. However, little is known about the initial stem cells in the yolk sac. We have isolated hematopoietic stem cells from early mouse embryonic yolk sac by using a sequential protocol of nonadherence to plastic, density gradient centrifugation, immunocytoadherence, and cell sorting. Isolated, nonadherent, density < 1.077 g/cm3, surface antigen AA4.1+, wheat germ agglutinin bright (WGAbright) cells give rise to multiple lineages, including T cells, B cells, and myeloid cells, as detected by using fetal thymus organ culture, S17 stromal feeder layers, or methylcellulose culture colony-forming cells, respectively. AA4.1+, WGAbright cells expressed high levels of heat-stable antigen (HSA) and CD45 (Ly-5) but did not significantly express major histocompatibility complex antigens, CD44, or Sca 1. Peak stem cell concentration is reached by day 11, before stem cells can be found in the liver, omentum, or thymus. In vivo long-term reconstitution of lethally irradiated mice was effected by as few as 720 AA4.1+, WGAbright yolk sac cells, but it required addition of a subset of bone marrow cells capable of providing immediate (short-term) radiation protection. Yolk sac donor-derived T cells, B cells, and macrophages were readily identified 6 months after transfer of yolk sac-derived stem cells. We suggest that, because of their cell surface phenotype as well as their capacity to differentiate in vitro and in vivo, the cells isolated from the mouse embryonic yolk sac may include the most primitive hematopoietic pluripotential stem cells yet identified. PMID- 8234266 TI - Serine-173 of the Epstein-Barr virus ZEBRA protein is required for DNA binding and is a target for casein kinase II phosphorylation. AB - An Epstein-Barr virus-encoded protein, ZEBRA, mediates the switch from latency to the viral lytic life cycle. ZEBRA's domain structure and DNA binding specificity resemble that of cellular transcriptional activators such as c-Fos/c-Jun. We show that ZEBRA, like c-Jun, is phosphorylated by casein kinase II (CKII). The principal site of phosphorylation is serine-173 (S173), five amino acids upstream of the basic DNA recognition domain. CKII phosphorylation abrogated ZEBRA's capacity to bind its target DNA sequences. S173 is a functional component of ZEBRA's DNA binding domain, since mutation of S173 to alanine (S173A) reduced DNA binding in vitro to 10% of wild-type levels. Transcriptional activation of a native viral promoter in vivo by mutant S173A was also reduced markedly. Reversible phosphorylation of S173 is likely to be an important means of regulating ZEBRA's activity in vivo. PMID- 8234267 TI - Stimulation of Trypanosoma cruzi adenylyl cyclase by an alpha D-globin fragment from Triatoma hindgut: effect on differentiation of epimastigote to trypomastigote forms. AB - A peptide from hindguts of the Triatoma hematophagous Chagas insect vector activates adenylyl cyclase activity in Trypanosoma cruzi epimastigote membranes and stimulates the in vitro differentiation of epimastigotes to metacyclic trypomastigotes. Hindguts were obtained from insects fed 2 days earlier with chicken blood. Purification was performed by gel filtration and HPLC on C18 and C4 columns. SDS/PAGE of the purified peptide showed a single band of about 10 kDa. The following sequence was determined for the 20 amino-terminal residues of this peptide: H2N-Met-Leu-Thr-Ala-Glu-Asp-Lys-Lys-Leu-Ile-Gln- Gln-Ala-Trp-Glu Lys-Ala-Ala-Ser-His. This sequence is identical to the amino terminus of chicken alpha D-globin. On a Western blot, the peptide immunoreacted with a polyclonal antibody against chicken globin D. A synthetic peptide corresponding to residues 1-40 of the alpha D-globin amino terminus also stimulated adenylyl cyclase activity and promoted differentiation. This 125I-labeled synthetic peptide bound specifically to T. cruzi epimastigote cells. Activation of epimastigote adenylyl cyclase by the hemoglobin-derived peptide may play an important role in T. cruzi differentiation and consequently in the transmission of Chagas disease. PMID- 8234268 TI - Probing the role of loop 2 in Ras function with unnatural amino acids. AB - The YDPT sequence motif (residues 32-35) in loop 2 (residues 32-40) of Ha-Ras p21 protein is conserved in the Ras protein family. X-ray crystal structures have revealed significant conformational differences in this region between the GTP- and GDP-bound forms. Moreover, mutations in this region block neoplastic transformation and prevent interaction with GTPase-activating protein (GAP), suggesting that this region may contribute to the effector function of Ras. To better understand the structural features required for GAP interaction and GTPase activity, the expanded repertoire of unnatural amino acid mutagenesis has been used to investigate the roles of the key residues, Pro-34, Thr-35, and Ile-36. A Pro-34-->methanoproline mutant, in which residue 34 is locked in the trans conformation, was found to retain high levels of intrinsic and GAP-activated GTPase activity, making unlikely conformational isomerization at this position. Deletion of a single methyl group from Ile (Ile-36-->norvaline) abolished GAP activation of Ras, revealing a remarkable specificity in this protein-protein interaction. Finally, replacement of Thr-35 with diastereomeric allo-threonine led to inactivation of Ras, demonstrating the importance of the orientation of this critical residue in Ras function. PMID- 8234269 TI - beta-Amyloid precursor protein binds to the neurite-promoting IKVAV site of laminin. AB - We previously characterized a 110-kDa membrane-associated laminin-binding protein (LBP110) from brain which binds the laminin A chain -Ile-Lys-Val-Ala-Val-(IKVAV) site and increases in injury. Here we demonstrate that antisera directed against different epitopes of beta-amyloid precursor protein (APP) recognize LBP110 and that APP is recognized by LBP110 antiserum. APP specifically binds IKVAV and not another biologically active laminin-derived peptide containing the amino acid sequence -Tyr-Ile-Gly-Ser-Arg-. PC-12 cells transfected with antisense APP RNA produce less APP and LBP110, and they form fewer processes when cultured on either laminin or the IKVAV peptide. Thus, LBP110 is a member of the APP family and a function for APP in neurite outgrowth is now defined. PMID- 8234270 TI - Blastocyst's state of activity determines the "window" of implantation in the receptive mouse uterus. AB - The present investigation studied the influence of the blastocyst's state of activity on the "window" of implantation in the receptive uterus in the mouse. The receptive state of the uterus is defined as the limited time when the uterine milieu is favorable to blastocyst acceptance and implantation. In the mouse, implantation occurs on day 4 (day 1 = vaginal plug). Ovariectomy in the morning of day 4 prior to preimplantation estrogen secretion results in blastocyst dormancy and delayed implantation. These conditions are maintained by continued progesterone (P4) treatment but can be terminated with an injection of estrogen leading to blastocyst activation and subsequent implantation. Blastocyst transfers into intact pseudopregnant mice demonstrated that the window of implantation on day 4 remains open at least through 1800 h for normal day 4 blastocysts but only up to 1400 h for dormant blastocysts. These results suggested that the blastocyst's state of activity influenced the normally operative window of implantation in the receptive uterus. This finding was further confirmed by inducing conditions of delayed implantation in pregnant donors and pseudopregnant recipients. They were ovariectomized on the morning of day 4 and maintained with daily injections of P4 from days 5 to 7. On day 7, dormant blastocysts from P4-treated delayed donors were transferred into the uteri of P4-treated delayed pseudopregnant recipients at 1, 2, 4, or 8 h after an injection of 17 beta-estradiol (E2). Dormant blastocysts transferred into delayed recipients at 1 h after E2 treatment resulted in implantation in most of the animals as compared to complete failure of blastocysts to implant after transfer to P4-treated delayed recipients at 4 or 8 h after E2 treatment. However, implantation did occur in P4-treated delayed recipients at these later hours of E2 treatment when the P4-treated delayed donors also received E2 prior to blastocyst transfer. Furthermore, the majority of day 4 normal blastocysts implanted when transferred into P4-treated delayed recipients even at 16 h after E2 treatment. Interestingly, day 7 dormant blastocysts cultured for 8 or 24 h for in vitro activation failed to implant after transfer to P4-treated delayed pseudopregnant recipients at 4 ir 8 h after E2 treatment, although they did implant after transfer at 1 h after E2 treatment. As expected, normal day 4 blastocysts failed to implant after transfer to P4-treated delayed pseudopregnant recipients. Thus, these results establish that the blastocyst's state of activity alters the timing of implantation (window) in the receptive uterus. Thus, the window for successful implantation could be defined as a limited time span when the activated stage of the blastocyst is superimposed on the receptive state of the uterus. This window remains open for a shorter period for dormant blastocysts than for a normal or dormant blastocysts after E2 activation. Furthermore, dormant blastocysts, which apparently achieved metabolic activation in vitro, failed to attain the same status as blastocysts activated in utero by E2 for implantation into the receptive uterus. A key finding of this investigation is that E2 induces very rapidly, but transiently (1 h), a factor(s) in the P4-primed uterus that activates the dormant blastocysts for implantation in the receptive uterus. PMID- 8234271 TI - Borrelia burgdorferi is clonal: implications for taxonomy and vaccine development. AB - The chromosomal genes fla and p93 and the ospA gene from a linear plasmid were sequenced from up to 15 isolates of Borrelia burgdorferi, which causes Lyme borreliosis in man. Comparison of the gene trees provides no evidence for genetic exchange between chromosomal genes, suggesting B. burgdorferi is strictly clonal. Comparison of the chromosomal gene trees with that of the plasmid-encoded ospA reveals that plasmid transfer between clones is rare. Evidence for intragenic recombination was found in only a single ospA allele. The analysis reveals three common clones and a number of rare clones that are so highly divergent that vaccines developed against one are unlikely to provide immunity to organisms from others. Consequently, an understanding of the geographic and genetic variability of B. burgdorferi will prove essential for the development of effective vaccines and programs for control. While the major clones might be regarded as different species, the clonal population structure, the geographic localization, and the widespread incidence of Lyme disease suggest that B. burgdorferi should remain the name for the entire array of organisms. PMID- 8234272 TI - Beet western yellows virus-associated RNA: an independently replicating RNA that stimulates virus accumulation. AB - Infections of plants by subviral RNA agents, alone or in association with virus genomic RNA molecules, are well known. The ST9 strain of beet western yellows virus encapsidates not only the 5.6-kilobase genomic RNA that is typical of luteoviruses, but also a 2.8-kilobase-associated RNA that has a distinct nucleotide sequence. The ST9-associated RNA has been postulated to be a satellite RNA, which by definition would be capable of replicating only in coinfections with beet western yellows virus or closely related viruses. To characterize the associated RNA, we inoculated protoplasts and leaves with in vitro transcripts of the virus genomic RNA and the ST9-associated RNA separately and in combination. Surprisingly, the ST9-associated RNA alone replicated efficiently in both protoplasts and leaves, and it stimulated accumulation of the virus genomic RNA in protoplasts. Thus, the ST9-associated RNA is a newly discovered type of plant infectious agent, which depends on its associated virus, beet western yellows virus, for encapsidation but not for replication. PMID- 8234273 TI - Bacteriophage T7 gene 2.5 protein: an essential protein for DNA replication. AB - The product of gene 2.5 of bacteriophage T7, a single-stranded DNA binding protein, physically interacts with the phage-encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phages that contain null mutants of gene 2.5 were constructed by homologous recombination. These gene 2.5 null mutants contain either a deletion of gene 2.5 (T7 delta 2.5) or an insertion into gene 2.5 and cannot grow in Escherichia coli (efficiency of plating, < 10(-8)). After infection of E. coli with T7 delta 2.5, host DNA synthesis is shut off, and phage DNA synthesis is reduced to < 1% of phage DNA synthesis in wild-type T7-infected E. coli cells as measured by incorporation of [3H]thymidine. In contrast, RNA synthesis is essentially normal in T7 delta 2.5 infected cells. The defects in growth and DNA replication are overcome by wild type gene 2.5 protein expressed from a plasmid harboring the T7 gene 2.5. PMID- 8234274 TI - Allelic instability in mitosis: a unified model for dominant disorders. AB - Recent findings indicate that tandemly repeated triplet sequences in certain disease-causing human genes may render these genes highly unstable not only in meiosis but also in mitosis. Typically, a dominant mutation arises upon expansion in the number of these repeated elements. We have considered how mitotic instability of this sort might affect both phenotypic expression and allele transmission. A model based on these considerations leads to the following predictions: (i) Phenotypic severity among individuals who inherit an unstable allele should be highly variable due to stochastic variation in the stage of its earliest mutagenic expansion. (ii) Strikingly increased severity or decreased age of onset in some offspring should arise because of parental germ-line mosaicism for an expanded or mutant allele. (iii) The magnitude of genetic anticipation should be more strongly correlated with paternal than with maternal age at the time of conception. (iv) Given a child born with a severe phenotype, the recurrence risk for a second severely affected child should be significantly elevated. (v) The severity of phenotype in a child should be positively correlated with that in a parent. Available data on fragile X syndrome, Huntington disease, and myotonic dystrophy are shown to be consistent with the model, and implications for an understanding of achondroplasia and other dominant disorders are discussed. PMID- 8234275 TI - Lymphocytes and macrophages outnumber oligodendroglia in normal fish spinal cord. AB - As shown by staining with a monoclonal antibody against fish CD45, leukocytes are present in very large numbers in the fish central nervous system. Their subtypes were distinguished by electron microscopy and found to include all major hematogenous forms except thrombocytes, the most numerous being tissue macrophages and lymphocytes. As a population, they differ fundamentally from ramified microglia, the restricted form of myeloid cells present in the central nervous system in mammals. They are rare in most grey matter regions but are concentrated in myelinated fiber tracts as well as in certain strata of the radial glial network. The macrophages engulf discarded myelin and outnumber the oligodendrocytes in normal spinal cord white matter, where the density of lymphocytes is > 5000-fold greater than reported in rat. PMID- 8234276 TI - Transcription factors NF-IL6 and NF-kappa B synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8. AB - Single binding sites for transcription factors NF-IL6 and NF-kappa B are present in the promoter of the interleukin (IL) 6 gene. Previous studies of internally deleted promoter mutants demonstrated that these two sites are important for the transcriptional regulation of this gene. In this report, we describe the synergistic activation of the IL-6 promoter by transcription factors NF-IL6 and NF-kappa B. Cotransfection of NF-IL6 with the NF-kappa B p65 subunit resulted in strong synergistic activation of an IL-6 promoter-reporter construct. Both the NF IL6 and NF-kappa B binding sites in the IL-6 promoter were required for synergistic activation. Similar synergistic activation was observed in the IL-8 promoter, which also contains both NF-IL6 and NF-kappa B binding sites. Furthermore, we demonstrated that NF-IL6 and the NF-kappa B p65 subunit directly associated via the basic leucine-zipper domain of NF-IL6 and the Rel homology domain of p65. Since the promoters of many other genes involved in the inflammatory and acute-phase responses also contain binding sites for NF-IL6 and NF-kappa B, the cooperation between these two factors may have an important role in these responses. We also discuss the possible interplay between various viral gene products and these two factors in the process of viral infection and constitutive cytokine production. PMID- 8234277 TI - On the role of macrophages in anthrax. AB - Bacillus anthracis, the causative agent of anthrax, produces systemic shock and death in susceptible animals, primarily through the action of its lethal toxin. This toxin, at high concentrations, induces lysis of macrophages in vitro but shows little or no effect on other cells. We found that when mice were specifically depleted of macrophages by silica injections, they became resistant to the toxin. Sensitivity could be restored by coinjection of toxin-sensitive cultured macrophages (RAW 264.7 cells) but not by coinjection of other cell lines tested. These results implied that macrophages mediate the action of lethal toxin in vivo and led us to investigate their role in death of the mammalian host. Sublytic concentrations of lethal toxin, orders of magnitude lower than those required to induce lysis of RAW 264.7 cells, were found to induce these cells to express interleukin 1 (IL-1) and tumor necrosis factor in vitro. Passive immunization against IL-1 or injection of an IL-1 receptor antagonist protected mice from toxin challenge, whereas anti-tumor necrosis factor provided little, if any, protection. These results imply that systemic shock and death from anthrax result primarily from the effects of high levels of cytokines, principally IL-1, produced by macrophages that have been stimulated by the anthrax lethal toxin. PMID- 8234278 TI - Mutant woodchuck hepatitis virus genomes from virions resemble rearranged hepadnaviral integrants in hepatocellular carcinoma. AB - Although hepadnaviruses are implicated in the etiology of hepatocellular carcinoma, the pathogenic mechanisms involved remain uncertain. Clonally propagated integrations of hepadnaviral DNA into cellular DNA can be demonstrated in most virally induced hepatocellular carcinomas. Integration occurs at random sites in cellular DNA, but the highly preferred sites in viral DNA are adjacent to the directly repeated sequence DR1, less often DR2, or in the cohesive overlap region. Integrants invariably contain simple deletions or complex rearrangements that have been thought to occur after integration. We report here the detection of mutant woodchuck hepatitis virus (WHV) genomes cloned from virions in serum that are strikingly similar to the rearranged hepadnaviral genomes found previously as integrated sequences in cellular DNA. Of 102 cloned genomes studied, 2 had large inverted duplications, 1 a 219-nucleotide direct duplication, and 1 a 219-nucleotide deletion. Virus-virus DNA junctions occurred either adjacent to DR1 or DR2 or in the cohesive overlap region at preferred topoisomerase I cleavage sites. Since these sites are located in the single stranded regions of the genome, cleavage by topoisomerase I would produce linear molecules that would be expected to be highly recombinogenic since this enzyme, possessing nicking and ligating activities, would remain covalently attached. Sucrose density gradient centrifugation coupled with polymerase chain reaction studies confirmed that the mutant WHV DNA forms resided in virions and did not represent free viral DNA released from infected cells or were unlikely to be an artifact of the cloning process. Thus, the finding in virions of mutant WHV DNA similar to WHV DNA integrated into cellular DNA suggests that the processes of mutation and integration are linked in some instances. Furthermore, the mutant genomes that are preferentially integrated into cellular DNA may have an etiologic role in hepatocarcinogenesis. PMID- 8234279 TI - Control of folding and membrane translocation by binding of the chaperone DnaJ to nascent polypeptides. AB - Recent evidence supports the view that cellular protein folding may be mediated by molecular chaperones. A fundamental question concerns the stage in its biogenesis at which the folding protein makes first contact with these components. We show here by crosslinking that the chaperone DnaJ binds nascent ribosome-bound polypeptide chains as short as 55 residues. Cotranslational binding of DnaJ to firefly luciferase and chloramphenicol acetyltransferase resulted in an arrest of folding as long as the functional partners of DnaJ in Escherichia coli, DnaK and GrpE, were missing. Protein uptake into microsomes and mitochondria was also interrupted by DnaJ. Both folding and post-translational translocation recommenced upon addition of DnaK and GrpE. We propose that DnaJ protects nascent polypeptide chains against aggregation and, in cooperation with Hsp70, controls their productive folding once a complete polypeptide or a polypeptide domain has been synthesized. PMID- 8234280 TI - Investigation of the validity of species status of Ixodes dammini (Acari: Ixodidae) using rDNA. AB - The two internal transcribed spacers (ITS1 and ITS2) of rDNA of three members of the Ixodes ricinus "complex" (Acari: Ixodidae) were sequenced. Sequence variation was assessed for the North American species I. scapularis, I. dammini, and I. pacificus at three levels: within individual/population, between individuals of different geographic origin within a species, and between species. Both spacers are highly variable, particularly with regard to small deletions and additions which may arise via replication slippage. Homogenization of rDNA multigene arrays for particular sequence variants appears to occur at a relatively rapid rate, since I. pacificus sequences differ from the others at numerous invariant sites, facilitating the use of these sequences to assess sibling species relationships. Based on maximum parsimony and two distance methods (unweighted pair-group with arithmetic means and neighbor-joining), sequence variation in ITS1 and ITS2 suggests that I. scapularis and I. dammini are not distinct species and that even individuals from geographically isolated locations are very similar. Individuals from geographically separated populations of I. pacificus appear to be relatively less closely related to each other but distinct from those of I. scapularis/dammini. In I. scapularis/dammini, diversity within and between individuals from geographic populations contributed equally to total sequence diversity. PMID- 8234281 TI - Cationic drug analysis using matrix-assisted laser desorption/ionization mass spectrometry: application to influx kinetics, multidrug resistance, and intracellular chemical change. AB - Highly sensitive and convenient analysis of intracellular cationic drugs has been achieved by applying matrix-assisted laser desorption/ionization mass spectrometry (MALD-MS). Tetraphenylphosphonium cation was readily identified and quantified (using methyltriphenylphosphonium cation as an internal standard) at subpicomole levels in crude lysate from < 4 x 10(3) FaDu human hypopharyngeal carcinoma cells. A quantitative MALD-MS time course for tetraphenylphosphonium cation accumulation into FaDu cells was comparable to a time course using scintillation counting with tritiated tetraphenylphosphonium. MALD-MS was also capable of demonstrating the reduced accumulation of the cationic drug rhodamine 123 by DoxR MCF7, a multiply drug-resistant human breast adenocarcinoma cell line, relative to the nonresistant parent line MCF7. In addition, MALD-MS was used to follow a chemical reaction inside intact FaDu cells: the formation of a hydrazone (II-51) from benzaldehyde and an acylhydrazide, 5-[tris(4 dimethylaminophenyl)phosphonio]pentanoyl hydrazide (II-25). These results suggest that MALD-MS may provide a rapid and practical alternative to existing methods for the analysis of cationic drugs, toxins, and their metabolites in cells and tissues. PMID- 8234282 TI - mu opiate receptor: cDNA cloning and expression. AB - mu opiate receptors recognize morphine with high affinity. A 2.1-kb rat brain cDNA whose predicted translation product displays 63% identity with recently described delta and kappa opiate receptor sequences was identified through polymerase chain reaction and cDNA homology approaches. This cDNA recognizes a 10.5-kb mRNA that is expressed in thalamic neurons. COS-cell expression confers naloxonazine-, Na(+)-, and GTP-sensitive binding of mu but not delta or kappa opioid ligands. Expressing cells bind morphine, [D-Ala2,N-methyl Phe4,glyol5]enkephalin (DAMGO), and [D-Ala2,D-Leu5]enkephalin (DADLE) with nanomolar or subnanomolar affinities, defining a mu opiate receptor that avidly recognizes analgesic and euphoric opiate drugs and opioid peptides. PMID- 8234283 TI - Atrial G protein-activated K+ channel: expression cloning and molecular properties. AB - Activity of several ion channels is controlled by heterotrimeric GTP-binding proteins (G proteins) via a membrane-delimited pathway that does not involve cytoplasmic intermediates. The best studied example is the K+ channel activated by muscarinic agonists in the atrium, which plays a crucial role in regulating the heartbeat. To enable studies of the molecular mechanisms of activation, this channel, denoted KGA, was cloned from a rat atrium cDNA library by functional coupling to coexpressed serotonin type 1A receptors in Xenopus oocytes. KGA displays regions of sequence homology to other inwardly rectifying channels as well as unique regions that may govern G-protein interaction. The expressed KGA channel is activated by serotonin 1A, muscarinic m2, and delta-opioid receptors via G proteins. KGA is activated by guanosine 5'-[gamma-thio]triphosphate in excised patches, confirming activation by a membrane-delimited pathway, and displays a conductance equal to that of the endogenous channel in atrial cells. The hypothesis that similar channels play a role in neuronal inhibition is supported by the cloning of a nearly identical channel (KGB1) from a rat brain cDNA library. PMID- 8234284 TI - Essential role of hypothalamic muscarinic and alpha-adrenergic receptors in atrial natriuretic peptide release induced by blood volume expansion. AB - Expansion of the blood volume induces natriuresis, which tends to return the blood volume to normal. This response is mediated at least in part by the release of atrial natriuretic peptide (ANP) into the circulation. Previous experiments have shown the participation of the anterior ventral third ventricular (AV3V) region of the hypothalamus in the ANP release that follows volume expansion. When injected into the AV3V region, the cholinergic drug carbachol induces natriuresis and the release of ANP. In the present experiments, microinjection of norepinephrine into the AV3V region induced natriuresis and an increase in plasma ANP. To determine whether cholinergic and alpha-adrenergic pathways are crucial to the volume expansion-induced release of ANP, certain receptor-blocking drugs were injected into the AV3V region of conscious rats. Thirty minutes later blood volume was expanded by intravenous injection of 2.0 ml/100 g of body weight of hypertonic saline (0.3 M NaCl). Microinjection of isotonic saline (2 microliters) into AV3V region of control animals 30 min prior to volume expansion had no effect on the 3-fold increase in plasma ANP concentrations measured 5 min after volume expansion. In contrast, although the receptor-blocking drugs did not alter the initial concentrations of plasma ANP 30 min later, just prior to volume expansion, blockade of muscarinic cholinergic receptors by intraventricular injection of 5 nmol (2 microliters) of atropine sulfate or methylatropine markedly reduced the response to volume expansion but did not obliterate it. Microinjection of the alpha receptor blocker phentolamine (5 nmol) into the AV3V 30 min prior to volume expansion also markedly suppressed the ANP response. Intraperitoneal (i.p.) injection of methylatropine (0.01 mmol/100 g of body weight), which does not cross the blood-brain barrier, also did not affect the basal levels of ANP 30 min after i.p. injection. But, in striking contrast with the blockade of the response to volume expansion induced by intraventricular injection of methylatropine, the response to volume expansion was markedly enhanced by i.p. injection of methylatropine. The results therefore indicate that hypothalamic muscarinic and alpha-adrenergic synapses are essential to release of ANP in response to volume expansion. These results are consistent with a hypothetical pathway for physiological control of ANP release which involves distension of baroreceptors within the right atria, carotid and aortic sinuses, and kidney which alters afferent input to brain stem noradrenergic neurons with axons projecting to the AV3V region. There they activate cholinergic interneurons by an alpha 1-adrenergic synapse. The cholinergic neurons in turn stimulate ANP neurons in this brain region via muscarinic receptors. The stimulation of these neurons activates efferent pathways which induce the release of ANP. PMID- 8234285 TI - Relationship between NMR transverse relaxation, trabecular bone architecture, and strength. AB - Structure, biomechanical competence, and incremental NMR line broadening (R'2) of water in the intertrabecular spaces of cancellous bone were examined on 22 cylindrical specimens from the lumbar vertebral bodies of 16 human subjects 24-86 years old (mean, 60 years old). A strong association (r = 0.91; P < 0.0001) was found between Young's modulus of elasticity and R'2 for a wide range of values corresponding to cancellous bone of very different morphologic composition. NMR line broadening is caused by the inhomogeneity of the magnetic field induced as a consequence of the coexistence of two adjacent phases of different diamagnetic susceptibility--i.e., mineralized bone and water in the marrow spaces. Structural analyses performed by means of NMR microscopy and digital image processing indicated that the variation in R'2 is closely related to the trabecular microstructure. Mean trabecular plate density measured along the direction of the magnetic field was found to play a major role in predicting R'2 (r = 0.74; P < 0.0001). This behavior was confirmed when the plate density was varied in individual specimens, which was achieved by rotating the specimen, making use of the bone's structural anisotropy. It is concluded that the NMR transverse relaxation rate in human cancellous bone of the spine is significantly determined by trabecular structural parameters relevant to biomechanical strength. The results further underscore the important role played by the transverse trabeculae in contributing to cancellous bone strength. The work has implications on possible in vivo use of quantitative magnetic resonance for the assessment of fracture risk in osteoporotic patients. PMID- 8234286 TI - Inhibition of endosome fusion by phospholipase A2 (PLA2) inhibitors points to a role for PLA2 in endocytosis. AB - Fusion of intracellular membrane-bound compartments is a common step in the transport of macromolecules along the endocytic and secretory pathways. A large number of factors active in the fusion process or its regulation have been identified; however, the actual sequence of events leading to membrane fusion is still unknown. In this study, we have assessed a possible role for PLA2 in endosome fusion by using an in vitro reconstitution assay and by examining endocytosis in intact cells. Several PLA2 inhibitors blocked endosome fusion in a broken-cell preparation. Inhibition was reversed by addition of arachidonic acid. At the electron microscope level, endosome clusters were observed even in the presence of inhibitors; however, actual fusion between endosomes was largely reduced. Fusion frequency increased upon the addition of arachidonic acid. A membrane-permeable PLA2 inhibitor blocked mixing of ligands internalized sequentially but did not affect internalization. The results indicate that vesicle fusion along the endocytic pathway requires a PLA2 activity. The effect of this activity would be, at least in part, mediated by arachidonic acid release. PMID- 8234287 TI - Differential expression of four members of the POU family of proteins in activated and phorbol 12-myristate 13-acetate-treated Jurkat T cells. AB - The POU family of proteins binds specifically to octamer DNA motifs present in the promoters of several genes and regulates their expression. We identified the presence of four members of the POU family of proteins, Oct-1, Oct-2, Oct-T1, and Oct-T2, in the human T-cell line Jurkat. To obtain insight into the physiological role played by these proteins in T cells, we studied the time course of expression of these genes in resting, activated, and phorbol 12-myristate 13 acetate (PMA)-treated cells. The expression of the gene encoding Oct-1 (now assigned the name OTF1 for octamer-binding transcription factor 1) remained unchanged and the levels of Oct-T2 mRNA decreased with increasing time of incubation to undetectable amounts in all three states of T-cell growth. The levels of Oct-2 mRNA and protein were increased in activated cells, were increased to a lesser extent in the PMA-treated cells, and were undetectable in resting cells. The levels of the Oct-T1 transcripts increased dramatically in PMA treated cells but not in resting or activated cells. Sequence analysis of the Oct T1 cDNA showed an open reading frame coding for a POU domain-containing protein of 42.7 kDa. Transient transfection of the gene encoding Oct-T1 decreased the activity of the interleukin 2 gene promoter in activated Jurkat cells. Further, there is evidence for an additional octamer-binding protein, Oct-T3, in Jurkat T cells. PMID- 8234288 TI - Tissue- and stratum-specific expression of the human involucrin promoter in transgenic mice. AB - Involucrin is a marker of keratinocyte terminal differentiation and is expressed only in the suprabasal layers of stratified squamous epithelium. In a previous study with various cell types in culture, we noted that expression of the putative human involucrin promoter was keratinocyte specific. To determine if this promoter is sufficient to direct expression to the suprabasal cells of stratified squamous epithelia in vivo, we have now generated transgenic mouse lines harboring the involucrin promoter sequences linked to a beta-galactosidase reporter gene. In the resulting lines, beta-galactosidase was expressed in the suprabasal compartment of stratified squamous epithelia and in hair follicles in a tissue-specific manner. In the palate, distinct vertical stacks of beta galactosidase-expressing cells were present, suggesting movement of clonally derived cells through the epithelium. The involucrin gene has a single intron upstream of the translational start site, and removal of this intron did not affect tissue- or stratum-specific expression. These results show that the 3.7-kb involucrin upstream sequences contain all the information necessary for a high level of tissue- and stratum-specific expression. PMID- 8234289 TI - An ets-related gene, ERG, is rearranged in human myeloid leukemia with t(16;21) chromosomal translocation. AB - The t(16;21)(p11;q22) translocation is a nonrandom chromosomal abnormality found in several types of myeloid leukemia, which show variable cytomorphological features. We constructed rodent-human somatic cell hybrids containing the der(16) chromosome from leukemic cells of a patient with t(16;21). Using these hybrids, we mapped the translocation breakpoint on the Not I restriction map of chromosome 21 which we had previously constructed. The result showed the proximity of the breakpoint to the ERG gene, a member of the ets oncogene superfamily. Polymerase chain reaction and Southern blot analyses of genomic DNA from the hybrids and from peripheral blood cells and bone marrow cells of patients with t(16;21) showed that the breakpoints were clustered within a single intron in the coding region of the ERG gene. This finding and the results obtained by Northern blot analysis suggested the formation of a chimeric product(s) by fusion of the ERG gene and an unknown counterpart gene on chromosome 16. PMID- 8234290 TI - G-protein alpha o subunit: mutation of conserved cysteines identifies a subunit contact surface and alters GDP affinity. AB - The reversible association of alpha and beta gamma subunits of GTP-binding proteins is important for signal transmission from a variety of cell-surface receptors to intracellular effectors. Previous work showed that 1,6 bis(maleimido)hexane, which crosslinks cysteine residues, crosslinks alpha o and alpha i-1 to beta gamma. These crosslinks are likely to form through a conserved cysteine because 1,6-bis(maleimido)hexane can also crosslink alpha i-2, alpha 1, alpha s and Drosophila alpha 1 to give products of the same apparent molecular weight as crosslinked alpha o beta gamma and alpha i-1 beta gamma. These proteins have only two cysteines in common. Therefore, we mutated each of the two conserved cysteines of alpha o to alanines. Mutation of Cys215 prevents crosslinking to beta gamma, but does not affect binding of guanosine 5'-[gamma thio]triphosphate or the ability of the mutated alpha subunit to bind beta gamma. In models of the alpha subunit based on the crystal structure of p21ras, Cys215 is located on the face opposite to the GTP-binding site and near an area that changes conformation depending on the nucleotide bound. This surface on the alpha subunit overlaps a putative effector binding region, raising important questions about the spatial organization of the proteins as they form ternary complexes. Mutation of Cys325 has no effect on crosslinking but, surprisingly, decreases by a factor of 10 the affinity of the mutated protein for GDP, relative to wild type, without changing the affinity for guanosine 5'-[gamma-thio]triphosphate. This mutation falls within a region thought to contact receptors and may represent a site through which receptors enhance the release of GDP. PMID- 8234291 TI - 26-10 Fab-digoxin complex: affinity and specificity due to surface complementarity. AB - We have determined the three-dimensional structures of the antigen-binding fragment of the anti-digoxin monoclonal antibody 26-10 in the uncomplexed state at 2.7 A resolution and as a complex with digoxin at 2.5 A resolution. Neither the antibody nor digoxin undergoes any significant conformational changes upon forming the complex. Digoxin interacts primarily with the antibody heavy chain and is oriented such that the carbohydrate groups are exposed to solvent and the lactone ring is buried in a deep pocket at the bottom of the combining site. Despite extensive interactions between antibody and antigen, no hydrogen bonds or salt links are formed between 26-10 and digoxin. Thus the 26-10-digoxin complex is unique among the known three-dimensional structures of antibody-antigen complexes in that specificity and high affinity arise primarily from shape complementarity. PMID- 8234292 TI - Split decomposition: a technique to analyze viral evolution. AB - A clustering technique allowing a restricted amount of overlapping and based on an abstract theory of coherent decompositions of finite metrics is used to analyze the evolution of foot-and-mouth disease viruses. The emerging picture is compatible with the existence of viral populations with a quasispecies structure and illustrates various forms of evolution of this virus family. In addition, it allows the correlation of these forms with geographic occurrence. PMID- 8234293 TI - Conformation-sensitive gel electrophoresis for rapid detection of single-base differences in double-stranded PCR products and DNA fragments: evidence for solvent-induced bends in DNA heteroduplexes. AB - Several techniques have recently been developed to detect single-base mismatches in DNA heteroduplexes that contain one strand of wild-type and one strand of mutated DNA. Here we tested the hypothesis that an appropriate system of mildly denaturing solvents can amplify the tendency of single-base mismatches to produce conformational changes, such as bends in the double helix, and thereby increase the differential migration of DNA heteroduplexes and homoduplexes during gel electrophoresis. The best separations of heteroduplexes and homoduplexes were obtained with a standard 6% polyacrylamide gel polymerized in 10% ethylene glycol/15% formamide/Tris-taurine buffer. As predicted by the hypothesis of solvent-induced bends, when the concentration of either ethylene glycol or formamide was increased, the differential migration decreased. Also, single-base mismatches within 50 bp of one end of a heteroduplex did not produce differential migration. Sixty of 68 single-base mismatches in a series of PCR products were detected in some 59 different sequence contexts. The eight mismatches not detected were either within 50 bp of the nearest end of the PCR product or in isolated high-melting-temperature domains. Therefore, it was possible to predict in advance the end regions and sequence contexts in which mismatches may be difficult to detect. The procedure can be applied to any PCR products of 200-800 bp and requires no special equipment or preparation of samples. PMID- 8234294 TI - Formation of functional peptide complexes of class II major histocompatibility complex proteins from subunits produced in Escherichia coli. AB - Class II major histocompatibility complex molecules play a major role in the immune response by binding peptide fragments of exogenous antigens and displaying them on the surfaces of antigen-presenting cells, where they can be recognized by T cells. To facilitate structural and functional studies of these molecules, we have produced truncated alpha and beta chains of the murine class II molecule I Ek in Escherichia coli (Ec-I-Ek) and have developed conditions to fold them in the presence of specific peptides with yields of complex approaching 2%. Reconstitution is specific since only unlabeled peptide known to bind I-Ek compete with biotinylated peptide, as assessed by ELISA. Complexes of the refolded heterodimer (Ec-I-Ek) with either of two different peptide antigens remain associated during nonreducing SDS/PAGE. Immobilized Ec-I-Ek-peptide complexes stimulate lymphokine production by three T-cell clones in an antigen specific manner with a dose-response relation comparable to previously described soluble I-Ek molecules produced in CHO cells. These results demonstrate that folding of Ek alpha and Ek beta polypeptides does not require any other protein to produce the biologically relevant conformation and that carbohydrate modification of this class II molecule is not necessary for alpha beta T-cell recognition. PMID- 8234295 TI - Fibroblasts can induce thymocyte positive selection in vivo. AB - During development in the thymus, thymocytes bearing alpha beta T-cell receptors are selected to mature if the receptors they bear are able to interact in some way with major histocompatibility complex (MHC) proteins expressed on thymic stromal cells. It has been shown that thymus cortical epithelial cells are usually the cells presenting the MHC molecules involved in this process of so called positive selection. Here we tested the ability of fibroblasts to mediate positive selection in vivo. Fibroblasts transfected with the genes for the MHC I Ab proteins were injected intrathymically into irradiated H-2k animals reconstituted with H-2bxk F1 fetal liver cells. Eight weeks later, the recipient mice were immunized and shown to contain peptide-specific I-Ab-restricted T cells. This demonstrates the ability of I-Ab-transfected fibroblasts to participate in positive selection. Thus a cell type that is not specialized to process and present antigens in the context of MHC class II molecules can mediate positive selection when transfected with an appropriate MHC molecule. The data also support the idea that the ability to mediate positive selection may not be limited to thymic cortical epithelium. PMID- 8234296 TI - An ethoxyquin-inducible aldehyde reductase from rat liver that metabolizes aflatoxin B1 defines a subfamily of aldo-keto reductases. AB - Protection of liver against the toxic and carcinogenic effects of aflatoxin B1 (AFB1) can be achieved through the induction of detoxification enzymes by chemoprotectors such as the phenolic antioxidant ethoxyquin. We have cloned and sequenced a cDNA encoding an aldehyde reductase (AFB1-AR), which is expressed in rat liver in response to dietary ethoxyquin. Expression of the cDNA in Escherichia coli and purification of the recombinant enzyme reveals that the protein exhibits aldehyde reductase activity and is capable of converting the protein-binding dialdehyde form of AFB1-dihydrodiol to the nonbinding dialcohol metabolite. We show that the mRNA encoding this enzyme is markedly elevated in the liver of rats fed an ethoxyquin-containing diet, correlating with acquisition of resistance to AFB1. AFB1-AR represents the only carcinogen-metabolizing aldehyde reductase identified to date that is induced by a chemoprotector. Alignment of the amino acid sequence of AFB1-AR with other known and putative aldehyde reductases shows that it defines a subfamily within the aldo-keto reductase superfamily. PMID- 8234297 TI - Arabidopsis thaliana auxotrophs reveal a tryptophan-independent biosynthetic pathway for indole-3-acetic acid. AB - We used tryptophan auxotrophs of the dicot Arabidopsis thaliana (wall cress) to determine whether tryptophan has the capacity to serve as a precursor to the auxin, indole-3-acetic acid (IAA). Quantitative gas chromatography-selected ion monitoring-mass spectrometry (GC-SIM-MS) revealed that the trp2-1 mutant, which is defective in the conversion of indole to tryptophan, accumulated amide- and ester-linked IAA at levels 38-fold and 19-fold, respectively, above those of the wild type. Tryptophan and free IAA were isolated from the trp2-1 mutant grown in the presence of [15N]anthranilate and [2H5]tryptophan, and the relative 15N and 2H5 enrichments of tryptophan and IAA were determined via GC-SIM-MS. The 15N enrichment of tryptophan, 13% +/- 4%, was less than the 15N enrichment of the IAA pool, 39% +/- 4%; therefore, IAA biosynthesis occurs via a tryptophan-independent pathway. The amount of 2H5 incorporated by the plant into IAA from tryptophan (9% +/- 4%) was low and only slightly above the level of spontaneous, nonenzymatic conversion of [2H5]tryptophan to [2H5]IAA. These results show that the dicot Arabidopsis is similar to the monocot Zea mays in that the major route of IAA biosynthesis does not occur through tryptophan. PMID- 8234298 TI - Cloning of cDNA for the alpha subunit of mouse insulin-like growth factor I receptor and the role of the receptor in metanephric development. AB - Various growth factors influence mammalian development by binding to specific cell surface receptors. These interactions are followed by a series of intracellular transductional events leading to a wide variety of biological effects. To establish the role of insulin-like growth factor I receptor (IGF-IR) in renal development, cDNA for the alpha subunit of the mouse IGF-IR was isolated, characterized, and used in expression studies and antisense experiments in a metanephric organ culture system. A 989-bp insert, encoding the signal peptide and 299 amino acids, isolated from a newborn mouse kidney cDNA library had 99% and 91% homology with the nucleotide sequences encoding the rat and the human IGF-IR, respectively. An approximately 11-kb message was readily detected by Northern blot analysis of RNA from the developing kidney at day 13 of gestation, and it declined during the subsequent embryonal and neonatal periods. In situ hybridization revealed high levels of message over the ureteric bud and its branches. A lower level of message was seen in the neonatal kidney, confined mainly to the tubules. Antisense oligodeoxynucleotide-treated metanephric kidneys were reduced in size and had a decreased population of nephrons with marked disorganization of ureteric bud branches. Immunofluorescence studies indicated an arrest of IGF-IR translation after antisense exposure. Immunoprecipitation studies showed a marked decrease in the biosynthesis of various extracellular matrix proteins that serve as regulators of morphogenesis. These studies suggest that the nucleotide sequence encoding the alpha subunit of mouse IGF-IR is highly conserved and that the receptor might play an essential role in the organogenesis of the kidney. PMID- 8234299 TI - Molecular cloning and characterization of the human A3 adenosine receptor. AB - The human A3 adenosine receptor was cloned from a striatal cDNA library using a probe derived from the homologous rat sequence. The cDNA encodes a protein of 318 amino acids and exhibits 72% and 85% overall identity with the rat and sheep A3 adenosine receptor sequences, respectively. Specific and saturable binding of the adenosine receptor agonist N6-(4-amino-3-[125I]iodobenzyl)adenosine [125I]ABA was measured on the human A3 receptor stably expressed in Chinese hamster ovary cells with a Kd = 10 nM. The potency order for adenosine receptor agonists was N ethylcarboxamidoadenosine (NECA) > or = (R)-N6-phenyl-2-propyladenosine [(R)-PIA] > N6-cyclopentyladenosine (CPA) > (S)-N6-phenyl-2-propyladenosine [(S)-PIA]. The human receptor was blocked by xanthine antagonists, most potently by 3-(3-iodo-4 aminobenzyl)-8-(4-oxyacetate)phenyl-1-propylxanthine (I-ABOPX) with a potency order of I-ABOPX > 1,3-dipropyl-8-(4-acrylate)phenylxanthine > or = xanthine amino congener >> 1,3-dipropyl-8-cyclopentylxanthine. Adenosine, NECA, (R)- and (S)-PIA, and CPA inhibited forskolin-stimulated cAMP accumulation by 30-40% in stably transfected cells; I-ABA is a partial agonist. When measured in the presence of antagonists, the dose-response curves of NECA-induced inhibition of forskolin-stimulated cAMP accumulation were right-shifted. Antagonist potencies determined by Schild analyses correlated well with those established by competition for radioligand binding. The A3 adenosine receptor transcript is widespread and, in contrast to the A1, A2a, and A2b transcripts, the most abundant expression is found in the lung and liver. The tissue distribution of A3 mRNA is more similar to the widespread profile found in sheep than to the restricted profile found in the rat. This raises the possibility that numerous physiological effects of adenosine may be mediated by A3 adenosine receptors. PMID- 8234300 TI - The maize transposable element system Ac/Ds as a mutagen in Arabidopsis: identification of an albino mutation induced by Ds insertion. AB - A two-component transposon system based on the Ac element of maize was used as a mutagen in Arabidopsis thaliana. Transposition of a Ds element marked with a hygromycin-resistance gene was activated from four different locations in the Arabidopsis genome. The progeny of 201 plants carrying independent transposition events were screened for mutants with severe, visible phenotypes. Seven mutants were identified and four of them were analyzed genetically. Three mutations were shown to be very closely linked to a transposed copy of the element. Moreover, a mutation (alb3) causing an albino phenotype was conclusively shown to be caused by insertion of the Ds element: somatic and germinal reversion of the mutation occurred in the presence of the transposase gene but not in its absence, and in three revertants the Ds had excised from its position in the mutant line. The DNA adjacent to Ds in the mutant was isolated and it was demonstrated that revertants retained part of the 8-bp duplication caused by insertion of Ds. These experiments indicate that the Ac/Ds system can be used as an insertional mutagen in the heterologous host Arabidopsis, which will permit the isolation of genes from this species by transposon tagging. PMID- 8234301 TI - Mapping of interacting domains between the nucleocapsid protein and the phosphoprotein of vesicular stomatitis virus by using a two-hybrid system. AB - Specific interaction between the nucleocapsid protein (N) and the phosphoprotein (P) of vesicular stomatitis virus (VSV), an important step in the life-cycle of the virus, was studied by using a two-hybrid system. Plasmids encoding P fused with the yeast GAL4 DNA-binding domain (pGALP) and N fused with the herpes simplex virus VP16 transactivating region (pVPN) were transfected into CHO cells along with a reporter plasmid encoding chloramphenicol acetyltransferase (CAT). The ability of N and P to associate in vivo was measured by activation of the CAT gene by the VP16 transactivating region. Transfection of plasmids pGALP and pVPN resulted in a high level of CAT activity, indicating that the N and P portions of the fusion proteins associated very strongly with each other. Progressive C terminal deletions of the P protein revealed two regions that are important for association with the N protein: the N-terminal acidic domain and the C-terminal basic domain. Phosphorylation of P protein was not required for N-P association. Various deletions and mutations of the N protein revealed the C-terminal 5 amino acids (Val-Glu-Phe-Asp-Lys), in particular the amino acids Val-Glu-Phe, to be critical for N association with P. This two-hybrid system can be used in other viral systems to study the interaction between proteins involved in transcription and replication. PMID- 8234302 TI - Central oxytocin inhibition of salt appetite in rats: evidence for differential sensing of plasma sodium and osmolality. AB - Sodium chloride ingestion is stimulated during conditions of sodium deficiency to maintain body fluid and electrolyte balance. Recent studies have indicated that salt appetite in rats is often inversely related to peripheral and central secretion of the hormone oxytocin (OT). We studied the potential role of central OT on salt and water ingestion by treating rats intracerebroventricularly with OT conjugated to the A chain of the plant cytotoxin ricin (rAOT) to produce a chronic selective inactivation of brain cells containing OT-receptive elements. The rats treated with rAOT and control rats treated with the ricin A chain alone were given 5-hr two-bottle (water and 0.5 M NaCl) drinking tests 30 min after they were made hyperosmolar by injections of hypertonic (2M) mannitol solution, which elevated plasma osmolality but reduced plasma Na+ concentration. In the control rats only water intake was stimulated in response to the induced hyperosmolality, but in the rAOT-treated rats hypertonic mannitol caused a robust salt appetite as well as thirst. Analogous results were obtained in rats treated with two different OT-receptor antagonists prior to induction of hyperosmolality with mannitol. In contrast to these results, when hyperosmolality was induced by administration of equivalently hypertonic (1M) NaCl, which elevated both plasma osmolality and plasma Na+ concentration, only water intake but not salt intake was stimulated in both control and OT-receptor antagonist-treated rats. When salt appetite was stimulated by the physiological stimulus of polyethylene glycol induced hypovolemia, hypertonic mannitol similarly inhibited salt ingestion in control animals but not in rAOT-treated rats, whereas hypertonic NaCl inhibited subsequent salt ingestion in both groups. These results suggest that salt appetite is regulated by both Na(+)- and osmolality-sensing mechanisms in rats. In addition, they indicate that central OT likely mediates a significant component of osmolality-related inhibition of salt appetite but does not appear to be essential for Na(+)-related inhibition of this important homeostatic behavior. PMID- 8234303 TI - Altered protein binding to the octamer motif appears to be an early event in programmed neuronal cell death. AB - Electrophoretic mobility-shift assays were used to characterize binding of nuclear proteins to consensus sequences for Sp1, E2F, octamer, and cAMP responsive enhancer element (CRE) during neuronal death in vitro after removal of nerve growth factor (NGF). Molecular events occurring prior to cell death in terminally differentiated PC12 cells could be divided into three phases: (i) within 2 hr of removing NGF, binding to the octamer sequence decreased, (ii) after 5-7 hr an increase in binding to CRE occurred; and (iii) after 14 hr (the point at which 50% of the cells are committed to die) a decrease in binding to the Sp1 sequence occurred. Assays performed with extracts from sympathetic ganglia indicated that changes in binding to CRE and octamer motifs also occurred during the period of developmental cell death in vivo. Double-stranded oligonucleotides were delivered to neurons to act as dominant negative "promoters" unable to couple to transcriptional events but capable of binding and sequestering transcription factors. Double-stranded but not single-stranded octamer oligonucleotides increased cell death of primary cultures of sympathetic neurons. Most of the induced neuronal cell death could be blocked with NGF, which is consistent with oligonucleotides activating an endogenous death program rather than having a nonspecific toxic effect. Other double-stranded oligonucleotides as well as a mutant octamer oligonucleotide had little or no effect on cell death. These data are consistent with the hypothesis that cell death results from a cascade of cellular and molecular events and that an early event in programmed neuronal cell death is a decrease in binding of transcription factor(s) to octamer motif sequences. PMID- 8234304 TI - Salmonella typhimurium induces membrane ruffling by a growth factor-receptor independent mechanism. AB - Invasive Salmonella typhimurium induces dramatic actin rearrangements on the membrane surface of mammalian cells as part of its entry mechanism. These changes, which are best characterized as membranous ruffles, closely resemble the membrane changes that occur when a growth factor binds to its receptor. Recently, inhibition of the function of the small GTPases rac and rho in quiescent serum starved fibroblasts was demonstrated to abolish growth factor-mediated ruffling and stress-fiber formation, respectively. In addition, actin changes induced by the oncogene ras were also shown to be regulated by rac and rho. Because Salmonella-induced actin rearrangements resemble those caused by growth factors, we investigated whether ras, rho, or rac regulates the membrane ruffling elicited by S. typhimurium. Surprisingly, inhibition of the functions of these GTPases had no effect on the ability of invasive S. typhimurium to induce membrane ruffles on a variety of tissue culture cells including Madin-Darby canine kidney cells, Swiss 3T3 fibroblasts, and Hep-2 cells. These results led us to examine the interactions of S. typhimurium with Henle-407 intestinal cells, which lack epidermal growth factor receptor on their membrane surface. We found no difference in the ability of invasive S. typhimurium to induce membrane ruffling and to enter Henle-407 cells with or without the epidermal growth factor receptor on the membrane surface. We, therefore, conclude that invasive S. typhimurium induces membrane ruffling and its own internalization by a rac-independent, growth factor-receptor-independent signaling pathway. PMID- 8234305 TI - Further evidence for function of the Drosophila Notch protein as a transmembrane receptor. AB - N locus mutations associated with unusual mutant phenotypes were found to alter the structure of the encoded protein. Two mutations, NCo and N60g11, eliminate much of the cytoplasmic domain. NCo can act as a null allele or as a competitive inhibitor of N+ function, whereas N60g11 produces dominant gain of function in some cell types. This difference in function can be attributed to retention of cdc10/SWI6 repeats in the Notch60g11 protein. The results suggest a role for these repeats in intracellular signaling and are consistent with action of Notch as a receptor. nd3 and l(1)NB alter extracellular epidermal growth factor-like and lin-12/Notch elements, respectively. nd3 eliminates a conserved cysteine residue, so the mutation may result in complete loss of function for a single Notch epidermal growth factor element. N60g11 and l(1)NB produce related gain-of function phenotypes. It is proposed that l(1)NB produces an extracellular modification of the protein that stimulates aberrant intracellular signaling by the Notch cytoplasmic domain. PMID- 8234306 TI - Formation of layer-specific fiber projections to the hippocampus in vitro. AB - The factors determining the layer-specific termination of hippocampal afferents are not known. Previous studies have suggested that the laminated termination of afferent fiber systems is caused by their sequential ingrowth during development. Here we have tested this temporal hypothesis of fiber segregation by an in vitro confrontation system in which the sequential arrival of entorhinal and commissural fibers was reversed. However, despite the temporal reversal of ingrowth, both fiber systems terminated in their normal positions. We conclude that the sequence of fiber ingrowth does not determine the lamination of hippocampal afferents. PMID- 8234307 TI - Structural characterization of the complete human perlecan gene and its promoter. AB - The complete intron-exon organization of the gene encoding human perlecan (HSPG2), the major heparan sulfate proteoglycan of basement membranes, has been elucidated, and specific exons have been assigned to coding sequences for the modular domains of the protein core. The gene was composed of 94 exons, spanning > 120 kbp of genomic DNA. The exon arrangement was analyzed vis-a-vis the modular structure of the perlecan, which harbors protein domains homologous to the low density lipoprotein receptor, laminin, epidermal growth factor, and neural cell adhesion molecule. The exon size and the intron phases were highly conserved when compared to the corresponding domains of the homologous genes, suggesting that most of this modular proteoglycan has evolved from a common ancestor by gene duplication or exon shuffling. The 5' flanking region revealed a structural organization characteristic of housekeeping and growth control-related genes. It lacked canonical TATA or CAAT boxes, but it contained several GC boxes with binding sites for the transcription factors SP1 and ETF. Consistent with the lack of a TATA element, the perlecan gene contained multiple transcription initiation sites distributed over 80 bp of genomic DNA. These results offer insights into the evolution of this chimeric molecule and provide the molecular basis for understanding the transcriptional control of this important gene. PMID- 8234308 TI - Notice of retraction. PMID- 8234309 TI - Nitric oxide: foe or friend to the injured brain? PMID- 8234310 TI - Images of science: science of images. PMID- 8234311 TI - Model observers for assessment of image quality. AB - Image quality can be defined objectively in terms of the performance of some "observer" (either a human or a mathematical model) for some task of practical interest. If the end user of the image will be a human, model observers are used to predict the task performance of the human, as measured by psychophysical studies, and hence to serve as the basis for optimization of image quality. In this paper, we consider the task of detection of a weak signal in a noisy image. The mathematical observers considered include the ideal Bayesian, the nonprewhitening matched filter, a model based on linear-discriminant analysis and referred to as the Hotelling observer, and the Hotelling and Bayesian observers modified to account for the spatial-frequency-selective channels in the human visual system. The theory behind these observer models is briefly reviewed, and several psychophysical studies relating to the choice among them are summarized. Only the Hotelling model with channels is mathematically tractable in all cases considered here and capable of accounting for all of these data. This model requires no adjustment of parameters to fit the data and is relatively insensitive to the details of the channel mechanism. We therefore suggest it as a useful model observer for the purpose of assessing and optimizing image quality with respect to simple detection tasks. PMID- 8234312 TI - Image processing: some challenging problems. AB - Image processing can be broadly defined as the manipulation of signals which are inherently multidimensional. The most common such signals are photographs and video sequences. The goals of processing or manipulation can be (i) compression for storage or transmission; (ii) enhancement or restoration; (iii) analysis, recognition, and understanding; or (iv) visualization for human observers. The use of image processing techniques has become almost ubiquitous; they find applications in such diverse areas as astronomy, archaeology, medicine, video communication, and electronic games. Nonetheless, many important problems in image processing remain unsolved. It is the goal of this paper to discuss some of these challenging problems. In Section I, we mention a number of outstanding problems. Then, in the remainder of this paper, we concentrate on one of them: very-low-bit-rate video compression. This is chosen because it involves almost all aspects of image processing. PMID- 8234313 TI - Color, contrast sensitivity, and the cone mosaic. AB - This paper evaluates the role of various stages in the human visual system in the detection of spatial patterns. Contrast sensitivity measurements were made for interference fringe stimuli in three directions in color space with a psychophysical technique that avoided blurring by the eye's optics including chromatic aberration. These measurements were compared with the performance of an ideal observer that incorporated optical factors, such as photon catch in the cone mosaic, that influence the detection of interference fringes. The comparison of human and ideal observer performance showed that neural factors influence the shape as well as the height of the foveal contrast sensitivity function for all color directions, including those that involve luminance modulation. Furthermore, when optical factors are taken into account, the neural visual system has the same contrast sensitivity for isoluminant stimuli seen by the middle-wavelength sensitive (M) and long-wavelength-sensitive (L) cones and isoluminant stimuli seen by the short-wavelength-sensitive (S) cones. Though the cone submosaics that feed these chromatic mechanisms have very different spatial properties, the later neural stages apparently have similar spatial properties. Finally, we review the evidence that cone sampling can produce aliasing distortion for gratings with spatial frequencies exceeding the resolution limit. Aliasing can be observed with gratings modulated in any of the three directions in color space we used. We discuss mechanisms that prevent aliasing in most ordinary viewing conditions. PMID- 8234314 TI - Color appearance: the effects of illumination and spatial pattern. AB - The color we perceive at each point in an image depends on information spread across the three spatial arrays of cone photoreceptors. I describe experiments aimed at clarifying how information is integrated across the spatial arrays to yield a color experience. We have found that changes of color appearance due to changes of the ambient illumination and the pattern's spatial frequency can be described by using a simple set of optical and neural transformations. Each transformation can be thought of as having two parts. First, the transformation converts the color representation into a new coordinate frame that is independent of the image contents. Second, the transformation scales the neural responses in the new coordinate frame by a gain factor that depends on the image contents. PMID- 8234315 TI - Nonlinear processes in visual pattern discrimination. AB - Visual performance on many simple pattern discrimination tasks can be accurately predicted by quasi-linear models composed of oriented linear filters followed by simple contrast nonlinearities. However, many complex discrimination tasks require highly nonlinear processes for their explanation. Evidence is provided for two nonlinear processes in pattern discrimination: (i) one process involves a sequence of filtering, rectification, and subsequent filtering to extract texture boundaries; (ii) the second process results from contrast gain-control processes. It is suggested that quasi-linear processes and nonlinear texture-boundary processes, each with an appropriate contrast gain control, may operate in parallel to provide the basis for all higher-level visual analyses. PMID- 8234316 TI - Issues of imaging science for future consideration. AB - Acceleration of the emergence of imaging science as a new discipline will require the development of new organizational structures to foster research and educational programs that integrate components of the traditional disciplines, all of which stand to benefit. However, the greatest impact of imaging science will likely be from computer-based general educational programs that present both visual and verbal materials utilizing software that is not only interactive but also analytic, diagnostic, and adaptive in response to individual students. Ultimately, this powerful learning paradigm will have profound effects on all aspects of our culture. Imaging science will not have emerged fully until the conceptual, organizational, educational, cultural, and ethical issues it raises have been addressed. PMID- 8234317 TI - Nitric oxide protects against cellular damage and cytotoxicity from reactive oxygen species. AB - Nitric oxide, NO, which is generated by various components of the immune system, has been presumed to be cytotoxic. However, NO has been proposed to be protective against cellular damage resulting during ischemia reperfusion. Along with NO there is often concomitant formation of superoxide/hydrogen peroxide, and hence a synergistic relationship between the cytotoxic effects of nitric oxide and these active oxygen species is frequently assumed. To study more carefully the potential synergy between NO and active oxygen species in mammalian cell cytotoxicity, we utilized either hypoxanthine/xanthine cell cytotoxicity, we utilized either hypoxanthine/xanthine oxidase (a system that generates superoxide/hydrogen peroxide) or hydrogen peroxide itself. NO generation was accomplished by the use of a class of compounds known as "NONOates," which release NO at ambient temperatures without the requirement of enzyme activation or biotransformation. When Chinese hamster lung fibroblasts (V79 cells) were exposed to hypoxanthine/xanthine oxidase for various times or increasing amounts of hydrogen peroxide, there was a dose-dependent decrease in survival of V79 cells as measured by clonogenic assays. However, in the presence of NO released from (C2H5)2N[N(O)NO]-Na+ (DEA/NO), the cytotoxicity resulting from superoxide or hydrogen peroxide was markedly abrogated. Similarly, primary cultures of rat mesencephalic dopaminergic cells exposed either to hydrogen peroxide or to hypoxanthine/xanthine oxidase resulted in the degradation of the dopamine uptake and release mechanism. As was observed in the case of the V79 cells, the presence of NO essentially abrogated this peroxide-mediated cytotoxic effect on mesencephalic cells. PMID- 8234318 TI - Subunit structure of a class A aspartate transcarbamoylase from Pseudomonas fluorescens. AB - The class A aspartate transcarbamoylase (ATCase, EC 2.1.3.2) from Pseudomonas fluorescens was purified to homogeneity with retention of full catalytic and regulatory functions. Careful determinations under conditions that minimized proteolysis showed that the molecule is a 1:1 stoichiometric complex of two polypeptide chains of 34 and 45 kDa. Pyridoxal phosphate is a competitive inhibitor of the enzyme (Ki = 1 microM). Reduction of the pyridoxal phosphate enzyme adduct with sodium boro[3H]hydride showed that the active site is located on the 34-kDa polypeptide. Affinity labeling with 5'-[p (fluorosulfonyl)benzoyl]adenosine, an ATP analog, suggested that the regulatory site is also located on the 34-kDa species. While the function of the 45-kDa subunit is unknown, neither carbamoyl phosphate synthetase nor dihydroorotase activities are associated with the ATCase. The molecular mass of the enzyme was determined by gel filtration, sedimentation velocity, and electron microscopy to be 464 kDa. Thus the enzyme is composed of six copies of the 34-kDa polypeptide and six copies of the 45-kDa polypeptide. The molecule has a Stokes' ratio of 70.9 A and a frictional ratio of 1.37, suggesting a compact globular shape. We propose that the P. fluorescens ATCase is composed of two trimers of 34-kDa catalytic chains and is likely to be a D3 dodecamer with an arrangement of subunits analogous to that of the class B ATCase molecules. PMID- 8234319 TI - Repair by human cell extracts of single (6-4) and cyclobutane thymine-thymine photoproducts in DNA. AB - One cis-syn cyclobutane thymine dimer or one (6-4) thymine-thymine photoproduct was built into an identical sequence of a closed-circular M13 duplex DNA, and nucleotide excision repair synthesis carried out by human cell extracts in the area containing each lesion was determined. Extracts from normal cells repaired the (6-4) photoproduct with a patch size of approximately 20-30 nucleotides, but repair was at least 10-fold lower at the cyclobutane dimer. The (6-4) lesion was repaired with comparable efficiency to a single acetylamino-fluorene-guanine adduct in a similar location. Extract from nucleotide excision repair-deficient xeroderma pigmentosum group A cells could not remove any of these adducts but could complete repair of the lesions after incision with Escherichia coli UvrABC proteins. This direct comparison of repair of two UV photoproducts, in an in vitro system where chromatin assembly and transcription are absent, suggests that the more rapid repair of the (6-4) lesion observed in the mammalian cell genome overall is due in part to a significant difference in the ability of the repair complex to locate and incise these lesions in DNA. PMID- 8234320 TI - Escherichia coli ribosomal protein L7/L12 dimers remain fully active after interchain crosslinking of the C-terminal domains in two orientations. AB - Cysteine site-directed mutagenesis was used to create variants of Escherichia coli ribosomal protein L7/L12 that have single cysteine substitutions, at residues 63 or 89, located in different exposed loops in the structure of the globular C-terminal domain indicated by the crystallographic structure. That structure shows a possible dimer interaction in which the two sites of cysteine substitution appear to be too distant for disulfide bond formation. After mild oxidation in solution both of the overexpressed purified cysteine-substituted proteins formed interchain disulfide crosslinked dimers in high yield. Both crosslinked dimers were fully active in restoring activity in poly(U)-directed polyphenylalanine synthesis to ribosomal core particles depleted of wild-type L7/L12. These results show that the two C-terminal domains have independent mobility. The activity of dimeric L7/L12 does not require the independent movement of the two globular C-terminal domains in an L7/L12 dimer; moreover, it appears independent of their mutual orientation when joined by crosslinking at the two loops. A third variant with a cysteine substitution at residue 33 near the junction between the alpha-helical N-terminal domain and the flexible hinge was prepared and tested. This protein was active in the protein synthesis assay in the reduced state. Oxidation produced the interchain crosslinked dimer in high yield, but this crosslinked dimer was inactive in polyphenylalanine synthesis. The inactivation was due to the inability of the Cys33-Cys33 oxidized dimer to bind to the core particle. PMID- 8234321 TI - Gene targeting of X chromosome-linked chronic granulomatous disease locus in a human myeloid leukemia cell line and rescue by expression of recombinant gp91phox. AB - The X chromosome-linked chronic granulomatous disease (X-CGD) locus, which encodes the gp91phox subunit of the phagocyte respiratory-burst oxidase cytochrome b, was disrupted by homologous recombination in the PLB-985 human myeloid cell line to develop an in vitro model of X-CGD. Superoxide formation was absent in targeted cells after differentiation to granulocytes but was rescued by stable transfection and expression of wild-type gp91phox cDNA. The targeted cell line should be useful in experiments aimed at defining functional regions within gp91phox by expression of mutant gp91phox cDNAs, complementing studies of naturally occurring mutations in X-CGD. In addition, the mutant line provides a model system in which to establish an experimental basis for the treatment of X CGD patients with gene replacement therapy. Rescued clones containing even modest amounts of recombinant gp91phox had respiratory-burst activity comparable to the wild-type PLB-985 line, suggesting that functional correction of X-CGD neutrophils may not require high-level expression of gp91phox. PMID- 8234322 TI - Local breathing and global unfolding in hydrogen exchange of barnase and its relationship to protein folding pathways. AB - We have measured the rate constants for exchange of amide protons in 15N-labeled wild-type barnase and a disulfide mutant that is more stable by 2 kcal.mol-1. The relative rate constants for exchange for wild type and mutant should reflect the changes in the equilibrium constants for local or global unfolding. The values for regions whose structure has been shown to be unaffected by the mutation fall into three subsets: those that are essentially unaffected by the mutation and so presumably exchange by local breathing; those where the energies change by close to 2 kcal.mol-1 and so presumably require global unfolding for exchange; and intermediate values that probably reflect a mixture of local and global unfolding in wild-type barnase. Amide protons that require the full change in unfolding energy are predominantly in the beta-sheet, which forms early in folding, but also include two that are involved in tertiary interactions that are known not to be formed until late in the folding pathway. Exchange in the major helix, which is known to form early, is largely unaffected by mutation and so exchanges by local breathing. There is thus no direct relationship between hydrogen-exchange behavior and the protein folding pathway. However, experiments on mutants of varying stability may provide further evidence on the sequence of events in folding. PMID- 8234323 TI - Use of a gene encoding a suppressor tRNA as a reporter of transcription: analyzing the action of the Nun protein of bacteriophage HK022. AB - The Nun protein of phage HK022 blocks the expression of genes that lie downstream of the nut sites of phage lambda. Nun is believed to act by promoting premature termination of transcription at or near these sites. To test this hypothesis and to facilitate mapping the sites of termination, we inserted a gene encoding a suppressor tRNA immediately downstream of the lambda nutL site and determined the effect of Nun on tRNA level. We found that Nun severely reduced the accumulation of mature, biologically active tRNA and promoted the accumulation of short, promoter-proximal transcripts whose 3' ends were dispersed over a 100-nucleotide region downstream of nutL. These results are consistent with the hypothesis that Nun terminates transcription within the region immediately downstream of nutL and are inconsistent with the hypothesis that the only action of Nun is to prevent translation of genes located downstream of the nut site. The stability, small size, and easily assayable biological function of suppressor tRNA recommend it as a reporter of transcription in other systems. PMID- 8234324 TI - Refined 1.8 A structure of human aldose reductase complexed with the potent inhibitor zopolrestat. AB - As the action of aldose reductase (EC 1.1.1.21) is believed to be linked to the pathogenesis of diabetic complications affecting the nervous, renal, and visual systems, the development of therapeutic agents has attracted intense effort. We report the refined 1.8 A x-ray structure of the human holoenzyme complexed with zopolrestat, one of the most potent noncompetitive inhibitors. The zopolrestat fits snugly in the hydrophobic active site pocket and induces a hinge-flap motion of two peptide segments that closes the pocket. Excellent complementarity and affinity are achieved on inhibitor binding by the formation of 110 contacts (< or = 4 A) with 15 residues (10 hydrophobic), 13 with the NADPH coenzyme and 9 with four water molecules. The structure is key to understanding the mode of action of this class of inhibitors and for rational design of better therapeutics. PMID- 8234325 TI - Crystal structure of a bacterial sialidase (from Salmonella typhimurium LT2) shows the same fold as an influenza virus neuraminidase. AB - Sialidases (EC 3.2.1.18 or neuraminidases) remove sialic acid from sialoglycoconjugates, are widely distributed in nature, and have been implicated in the pathogenesis of many diseases. The three-dimensional structure of influenza virus sialidase is known, and we now report the three-dimensional structure of a bacterial sialidase, from Salmonella typhimurium LT2, at 2.0-A resolution and the structure of its complex with the inhibitor 2-deoxy-2,3 dehydro-N-acetylneuraminic acid at 2.2-A resolution. The viral enzyme is a tetramer; the bacterial enzyme, a monomer. Although the monomers are of similar size (approximately 380 residues), the sequence similarity is low (approximately 15%). The viral enzyme contains at least eight disulfide bridges, conserved in all strains, and binds Ca2+, which enhances activity; the bacterial enzyme contains one disulfide and does not bind Ca2+. Comparison of the two structures shows a remarkable similarity both in the general fold and in the spatial arrangement of the catalytic residues. However, an rms fit of 3.1 A between 264 C alpha atoms of the S. typhimurium enzyme and those from an influenza A virus reflects some major differences in the fold. In common with the viral enzyme, the bacterial enzyme active site consists of an arginine triad, a hydrophobic pocket, and a key tyrosine and glutamic acid, but differences in the interactions with the O4 and glycerol groups of the inhibitor reflect differing kinetics and substrate preferences of the two enzymes. The repeating "Asp-box" motifs observed among the nonviral sialidase sequences occur at topologically equivalent positions on the outside of the structure. Implications of the structure for the catalytic mechanism, evolution, and secretion of the enzyme are discussed. PMID- 8234326 TI - Analysis of somatic hypermutation in mouse Peyer's patches using immunoglobulin kappa light-chain transgenes. AB - We have exploited mice transgenic for an immunoglobulin kappa light chain in order to show that immunoglobulin genes in the B cells of Peyer's patches in unimmunized mice carry a high level of somatic mutations. Most of the mutations are found in the subpopulation of B cells which, based on peanut agglutinin binding, derive from the germinal centers. The number of mutations per clone and their distribution along the variable gene segment (indicative of untemplated point mutations) are very similar to those found in antigen-specific splenic B cells of normal mice after secondary immunization. The mutations accumulate mainly in complementarity-determining region 1, in particular in some specific codons (Ser-26, Ser-31, and Ser-77) which have been previously recognized as intrinsic hypermutational hotspots. These results suggest that, as in the spleen, somatic mutation occurs in B cells which have migrated to the germinal centers, probably as a consequence of stimulation by antigens present in the gut environment. Transgenic animals are increasingly being used to define the signals involved in hypermutation. However, their subsequent study is very time-consuming because it is based on immunization and analysis of hybridomas or antigen selected cells. We propose that the use of Peyer's patches of unimmunized adult mice offers a reliable and simple approach to analyze hypermutation of transgenes. PMID- 8234327 TI - Gene sequence tags from Plasmodium falciparum genomic DNA fragments prepared by the "genease" activity of mung bean nuclease. AB - A genes-first approach to genome sequencing is described which efficiently generates gene sequence tags from genomic DNA. Mung bean nuclease (EC 3.1.30.1) cleaves the genomic DNA of many organisms before and after genes and within some introns. Analysis of gene sequence tags prepared from mung bean nuclease-digested Plasmodium falciparum DNA demonstrates that this method has several advantages over the popular cDNA expressed sequence tag approach. To date, 673 sequence tags containing over 215 kb of sequence have been generated from 400 clones. Sixty clones (15%) have significant similarity to sequences in the protein and translated nucleic acid data bases. These represent 51 unique genes, of which only 5 encode previously known P. falciparum proteins. The identified proteins include those expressed in erythrocytic, exoerythrocytic, and gametocytic stages of the parasite. Thirty percent of clones identified appear to carry complete coding regions. The spacer DNA separating genes is rarely cloned. These gene sequence tags will form a useful data base from which to initiate projects to develop new therapeutics, vaccines, and strategies to control human malaria. PMID- 8234328 TI - Specific prolongation of allograft survival by a T-cell-receptor-derived peptide. AB - Allograft rejection results from the specific recognition by host CD8+ T cells of allogeneic major histocompatibility complex (MHC) molecules on the tissue graft. The specificity of this cellular response is determined by the molecular interaction of the T-cell receptor (TCR) on host T cells with the MHC molecule and its bound ligand on the grafted tissue. To better understand the precise manner by which the TCR interacts with the MHC-peptide complex and how to therapeutically intervene, we have studied the allogeneic response to the mouse class I MHC molecule Ld. In this report, the therapeutic potential of a synthetic peptide derived from the TCR V beta 8 variable region that predominates in responses to Ld was tested. This V beta 8-derived peptide was found to dramatically and specifically block the in vivo and in vitro allogeneic response to Ld. Furthermore, this specific blocking is not dependent upon the presence of V beta 8+ effector cells nor does the V beta 8 peptide bind to the Ld ligand binding cleft. We propose that this peptide functions as an antagonist, competing with the native TCR for recognition of the Ld molecule. PMID- 8234329 TI - Mutants defective in the energy-conserving NADH dehydrogenase of Salmonella typhimurium identified by a decrease in energy-dependent proteolysis after carbon starvation. AB - NADH dehydrogenase is the first component of the respiratory chain. It transfers electrons from NADH to ubiquinone and concomitantly establishes a proton motive force across the membrane. Salmonella typhimurium mutants defective in this enzyme were isolated in a screen for strains with increased expression of beta galactosidase from a hemA-lacZ protein fusion. This unexpected phenotype results from stabilization of the hybrid protein during carbon starvation and is apparently due to an energy requirement for proteolytic attack. Sequence analysis of DNA fragments cloned from an insertion mutant indicates that S. typhimurium has a large cluster of genes encoding the energy-conserving NADH dehydrogenase, similar to one recently described in Paracoccus denitrificans. These findings establish the potential for genetic analysis of a complex enzyme whose function, especially in proton efflux, is poorly understood. PMID- 8234330 TI - Genomic clone for sandbar shark lambda light chain: generation of diversity in the absence of gene rearrangement. AB - While the general structure of immunoglobulin chains has remained relatively unchanged throughout evolution, the organization of the genes encoding these molecules differs substantially. To understand how the rearranging immunoglobulin system arose, it is necessary to examine living representatives of the most early vertebrate phyla. Elasmo-branches, which include the sharks, skates, and rays, are the most primitive phylogenetic class of vertebrates from which immunoglobulin DNA sequences have been obtained. In the sandbar shark (Carcharhinus plumbeus), the genes are arranged in individual clusters in which a single variable (V), joining (J), and constant (C) region gene, along with upstream regulatory elements, span a distance of approximately 4.4 kb or approximately 5.8 kb. We report the complete sequence of a genomic clone encoding sandbar shark lambda light chain. A unique finding of our study is that the V and J genes are fused in the germ line. Three additional clones have been shown by DNA sequencing to also have fused V and J genes. The four clones have complementarity-determining regions 3 of various lengths and amino acid sequence variability similar to the products of rearranged genes. Furthermore, analysis by polymerase chain reaction technology revealed an additional 26 genomic clones demonstrating fusion of the V and J segments. Therefore, VJ fusion is the prominent organizational feature of sandbar shark immunoglobulin light chain genes. This finding raises questions concerning the necessity of recombination to produce an antibody repertoire capable of reacting against a diverse array of antigens. PMID- 8234331 TI - Occurrence of fragmented 16S rRNA in an obligate bacterial endosymbiont of Paramecium caudatum. AB - The phylogenetic position of Caedibacter caryophila, a so far noncultured killer symbiont of Paramecium caudatum, was elucidated by comparative sequence analysis of in vitro amplified 16S rRNA genes (rDNA). C. caryophila is a member of the alpha subclass of the Proteobacteria phylum. Within this subclass C. caryophila is moderately related to Holospora obtusa, which is another obligate endosymbiont of Paramecium caudatum, and to Rickettsia. A 16S rRNA targeted specific hybridization probe was designed and used for in situ detection of C. caryophila within its host cell. Comparison of the 16S rDNA primary structure of C. caryophila with homologous sequences from other bacteria revealed an unusual insertion of 194 base pairs within the 5'-terminal part of the corresponding gene. The intervening sequence is not present in mature 16S rRNA of C. caryophila. It was demonstrated that C. caryophila contained fragmented 16S rRNA. PMID- 8234332 TI - Localized 1H NMR measurement of glucose consumption in the human brain during visual stimulation. AB - Spatially localized 1H NMR spectroscopy has been applied to measure changes in brain glucose concentration during 8-Hz photic stimulation. NMR spectroscopic measurements were made in a 12-cm3 volume centered on the calcarine fissure and encompassing the primary visual cortex. The average maximum change in glucose levels was 0.34 mumol.g-1 (n = 5) at 15 min; glucose level had turned toward resting level at 25 min. The glucose change was used to calculate the increase of glucose cerebral metabolic rate in the visual cortex region for individual subjects by using the Michaelis-Menten model of glucose transport on the assumption of constant transport kinetics. The glucose cerebral metabolic rate was calculated to increase over the nonstimulated rate by 22% during the first 15 min of photic stimulation. A model in which the glucose metabolic rate gradually decreases during stimulation was proposed as a possible explanation for the recovery of brain glucose and previously measured lactate concentrations to prestimulus values after 15 min. PMID- 8234333 TI - Antisense inhibition of the p65 subunit of NF-kappa B blocks tumorigenicity and causes tumor regression. AB - The NF-kappa B transcription factor, composed of two proteins, p50 and p65, is a pleiotropic activator that participates in the induction of a wide variety of cellular genes. Various cell adhesion molecules have NF-kappa B binding sites and may play an important role in inflammatory response, tumorigenicity, and metastasis. In an earlier study, we demonstrated that adhesion of diverse transformed cells was blocked by antisense inhibition of the p65 subunit of NF kappa B. Since cell-substratum interactions play an important role in tumorigenicity, we reasoned that antisense p65 could inhibit tumorigenicity. In diverse transformed cell lines, phosphorothioate antisense oligonucleotides to p65 inhibited in vitro growth, reduced soft-agar colony formation, and eliminated the ability of cells to adhere to an extracellular matrix. Stable transfectants of a fibrosarcoma cell line expressing dexamethasone-inducible antisense RNA to p65 showed inhibition of in vitro growth and in vivo tumor development. In response to inducible expression of antisense RNA, a pronounced tumor regression was seen in nude mice. The administration of antisense but not sense p65 oligonucleotides caused a pronounced inhibition of tumorigenicity in nude mice injected with diverse tumor-derived cell lines. Inhibitors of NF-kappa B function may thus be useful in the treatment of cancer. PMID- 8234334 TI - Leucine aminopeptidase: an inducible component of the defense response in Lycopersicon esculentum (tomato). AB - A leucine aminopeptidase (EC 3.4.11.1) cDNA clone (DR57) that was induced in response to Pseudomonas syringae pv. tomato (P.s. tomato) infection was isolated using a subtractive hybridization-enriched cDNA probe. Genomic DNA blot analysis showed that the tomato genome had two leucine aminopeptidase genes. The levels of DR57 mRNAs after P.s. tomato infection and mechanical wounding were determined in two inbred tomato lines that exhibit susceptibility and resistance to P.s. tomato. DR57 mRNAs were detected 12 hours after infection and 4 hours after wounding. Furthermore, DR57 mRNAs were systemically induced in response to wounding. DR57 mRNAs were induced in leaves after Spodoptera littoralis feeding but were not detected in detached leaf controls. Possible roles for the DR57 leucine aminopeptidase in the defense reactions are discussed. PMID- 8234335 TI - Coding coenzyme handles: a hypothesis for the origin of the genetic code. AB - The coding coenzyme handle hypothesis suggests that useful coding preceded translation. Early adapters, the ancestors of present-day anticodons, were charged with amino acids acting as coenzymes of ribozymes in a metabolically complex RNA world. The ancestral aminoacyl-adapter synthetases could have been similar to present-day self-splicing tRNA introns. A codon-anticodon discriminator base complex embedded in these synthetases could have played an important role in amino acid recognition. Extension of the genetic code proceeded through the take-over of nonsense codons by novel amino acids, related to already coded ones either through precursor-product relationship or physicochemical similarity. The hypothesis is open for experimental tests. PMID- 8234336 TI - Mutations that alter the third cytoplasmic loop of the a-factor receptor lead to a constitutive and hypersensitive phenotype. AB - The STE3 gene of Saccharomyces cerevisiae encodes a G protein-coupled receptor that is specific for the mating pheromone a-factor. The ste3L194Q mutation, which leads to the substitution of glutamine for leucine-194 within the third cytoplasmic loop of the receptor, resulted in a 20-fold increase in pheromone sensitivity and also caused partial constitutive activation of the response pathway. Moreover, other amino acid substitutions at the 194 position and several deletion mutations that collectively remove most of the third cytoplasmic loop resulted in hyperactive receptors. Therefore, we suggest that one role of the third cytoplasmic loop is to function as a negative regulatory domain involved in the maintenance of a nonsignaling state of the receptor. The constitutive activity and the pheromone hypersensitivity of ste3L194Q cells were recessive, suggesting that the wild-type receptor can antagonize the signal associated with the activated receptor. The ste3 delta 306 mutation, which results in truncation of most of the C-terminal domain of the receptor, led to a 20-fold increase in pheromone sensitivity, indicating that this domain also mediates negative regulation of the receptor. The ste3L194Q and ste3 delta 306 mutations appear to affect receptor activity independently, because the double mutant was associated with a 400-fold increase in pheromone sensitivity. PMID- 8234337 TI - Genetic evidence for the roles of the bud-site-selection genes BUD5 and BUD2 in control of the Rsr1p (Bud1p) GTPase in yeast. AB - Yeast cells normally display either an axial (for MATa or MAT alpha cells) or bipolar (for MATa/alpha cells) pattern of bud-site selection. The RSR1 gene, which was previously identified as a multicopy suppressor of Ts- mutations in the bud-emergence gene CDC24, encodes a GTPase of the Ras family that is required for both budding patterns. Mutations in Rsr1p that presumably block its ability to bind or hydrolyze GTP cause a randomized budding phenotype, suggesting that regulators of Rsr1p will prove to be required for proper bud positioning. The BUD5 gene product is required for proper bud-site selection and contains similarity to GDP-dissociation stimulators (GDS) for Ras-type proteins, suggesting that Bud5p may be a GDS for Rsr1p. Here I report that BUD5 is required for wild-type RSR1, but not for mutationally activated rsr1val12, to serve as a multicopy suppressor of cdc24, indicating that Bud5p functions as a GDS for Rsr1p in vivo. To identify the GAP (GTPase-activating protein) for Rsr1p, a genetic selection was designed based on the observation that mutationally activated rsr1val12, but not wild-type RSR1, can serve as a multicopy suppressor of yeast RAS2(Ts) mutants. Mutants were selected that allowed wild-type RSR1 to act as a multicopy suppressor of RAS2(Ts). Two such mutations proved to be in the BUD2 gene, suggesting that Bud2p functions as a GAP for Rsr1p in vivo. PMID- 8234338 TI - Preformed dimeric state of the sensor protein VirA is involved in plant- Agrobacterium signal transduction. AB - Plant signal molecules such as acetosyringone and certain monosaccharides induce the expression of Agrobacterium tumefaciens virulence (vir) genes, which are required for the processing, transfer, and possibly integration of a piece of the bacterial plasmid DNA (T-DNA) into the plant genome. Two fo the vir genes, virA and virG, belonging to the bacterial two-component regulatory system family, control the induction of vir genes by plant signals. virA encodes a membrane bound sensor kinase protein and virG encodes a cytoplasmic regulator protein. Although it is well established from in vitro studies that the signal transduction process involves VirA autophosphorylation and subsequent phosphate transfer to VirG, the structural state of the VirA protein involved in signal transduction is not understood. In this communication, we describe an in vivo crosslinking approach which provides physical evidence that VirA exists as a homodimer in its native configuration. The dimerization of VirA neither requires nor is stimulated by the plant signal molecule acetosyringone. We also present genetic data which support the hypothesis that VirA exists as a homodimer which is the functional state transducing the plant signal in an intersubunit mechanism. To our knowledge, this report provides the first evidence that a bacterial membrane-bound sensor kinase exists and functions as a homodimer in vivo. PMID- 8234339 TI - Transforming growth factor beta 1 (TGF-beta 1) controls expression of major histocompatibility genes in the postnatal mouse: aberrant histocompatibility antigen expression in the pathogenesis of the TGF-beta 1 null mouse phenotype. AB - The phenotype of the transforming growth factor beta 1 (TGF-beta 1) null mouse has been previously described and is characterized by inflammatory infiltrates in multiple organs leading to a wasting syndrome and death as early as 3 weeks after birth. Since this phenotype occurs in the absence of any detectable pathogen, potential autoimmune disease mechanisms were investigated. We examined major histocompatibility complex (MHC) mRNA expression in tissues of the TGF-beta 1 null mouse and found levels of both the class I and class II MHC mRNA elevated compared to normal or TGF-beta 1 heterozygous littermates. This elevated expression was seen prior to any evidence of inflammatory infiltrates, suggesting a causal relationship between increased MHC expression and activation of immune cell populations. Cell surface expression of MHC molecules was detected by immunohistochemistry and correlated well with mRNA levels. Expression of mRNA for interferon gamma and its receptor was unchanged at the ages when increased MHC expression became apparent. Down-regulation of class I MHC expression by TGF-beta 1 was also demonstrated in vitro in fibroblasts isolated from TGF-beta 1 null mice. These findings suggest that one natural function of TGF-beta 1 is to control expression of both MHC classes. Altered regulation of MHC expression may be a critical step leading to the multifocal inflammation and wasting syndrome seen in the TGF-beta 1 null mouse. These results suggest potential applications for TGF-beta in the management of autoimmune disease, allograft rejection, and other problems associated with altered MHC expression. PMID- 8234340 TI - Three-dimensional model for the hormone binding domains of steroid receptors. AB - We have used a motif-based structural search method to identify structural homologs of the hormone binding domains of the nuclear receptors from among a set of known protein structures and have found the closest similarity with members of the subtilisin-like serine proteases. These proteins consist of an open twisted sheet of parallel beta-strands flanked on both sides by alpha-helices. The alignment with the protease scaffold was refined by using multiple sequence prealignment of different sets of nuclear receptors, and alternative model structures were screened by considering their consistency with the results of biochemical experiments defining the ligand binding pocket. In the most favored model, nearly all of the residues thought to be involved in ligand binding map to a pocket of appropriate dimensions where the subtilisin-like proteases have their active site. The three-dimensional model that we propose for the hormone binding domains of the nuclear receptors provides a framework for the design of experiments to further investigate nuclear receptor structure and function. PMID- 8234341 TI - Cloning and pharmacological characterization of a rat kappa opioid receptor. AB - A full-length cDNA was isolated from a rat striatal library by using low stringency screening with two PCR fragments, one spanning transmembrane domains 3 6 of the mouse delta opioid receptor and the other unidentified but homologous to the mouse delta receptor from rat brain. The novel cDNA had a long open reading frame encoding a protein of 380 residues with 59% identity to the mouse delta receptor and topography consistent with a seven-helix guanine nucleotide-binding protein-coupled receptor. COS-1 cells transfected with the coding region of this clone showed high-affinity binding to kappa opioid receptor-selective ligands such as dynorphin A and U-50,488 and also nonselective opioid ligands such as bremazocine, ethylketocyclazocine, and naloxone. Not bound at all (or bound with low affinity) were dynorphin A-(2-13), enantiomers of naloxone and levophanol [i.e., (+)-naloxone and dextrorphan], and selective mu and delta opioid receptor ligands. Activation of the expressed receptor by kappa receptor agonists led to inhibition of cAMP. Finally, in situ hybridization revealed a mRNA distribution in rat brain that corresponded well to the distribution of binding sites labeled with kappa-selective ligands. These observations indicate that we have cloned a cDNA encoding a rat kappa receptor of the kappa 1 subtype. PMID- 8234342 TI - Formation of the active site of ribulose-1,5-bisphosphate carboxylase/oxygenase by a disorder-order transition from the unactivated to the activated form. AB - Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyzes the key first step in photosynthetic CO2 fixation, the reaction that incorporates CO2 into sugar. In this study, refined crystal structures of unactivated tobacco RuBisCO and activated RuBisCO from spinach and tobacco, in complex with the reaction intermediate analog 2-carboxyarabinitol 1,5-bisphosphate (CABP), are compared. Both plant enzymes are hexadecameric complexes of eight large and eight small subunits with a total relative molecular mass of approximately 550,000. The comparison of activated and unactivated forms of RuBisCO provides insight into the dynamics of action of this enzyme. The catalytic site, which is open to the solvent in the unactivated enzyme, becomes shielded in the activated CABP complex. This shielding is accomplished by a 12-A movement of the active-site "loop 6" (residues 331-338) and a disorder-order transition of three loops near the active-site entrance, the N terminus, the C terminus, and a loop comprising residues 64-68. All these residues belong to the catalytic large subunit. Domain rotations of about 2 degrees are observed, also tightening the active-site cleft. These observations provide an explanation for the extremely tight binding (Kd < or = 10(-11) M) of the CABP molecule. A striking correlation exists between crystallographic temperature factors in the activated enzyme and the magnitude of the atomic movement upon activation. PMID- 8234343 TI - Rapid interchange between two distinct forms of cyanobacterial photosystem II reaction-center protein D1 in response to photoinhibition. AB - We have studied photoinhibition of photosynthesis in the cyanobacterium Synechococcus sp. PCC 7942, which possesses two distinct forms of the photosystem II reaction-center protein D1 (D1:1 and D1:2). We report here that when cells adapted to a growth irradiance of 50 mumol.m-2.s-1 are exposed to an irradiance of 500 mumol.m-2.s-1, the normally predominant D1 form (D1:1) is rapidly replaced with the alternative D1:2. This interchange is not only complete within the first hour of photoinhibition but is also fully reversible once cells are returned to 50 mumol.m-2 x s-1. By using a mutant that synthesizes only D1:1, we show that the failure to replace D1:1 with D1:2 during photoinhibition results in severe loss of photosynthetic activity as well as a diminished capacity to recover after the stress period. We believe that this interchange between D1 forms may constitute an active component in a protection mechanism unique among photosynthetic organisms that enables cyanobacteria to effectively cope with and recover from photoinhibition. PMID- 8234344 TI - Cell-specific action and mutable structure of a transcription factor effector domain. AB - POU proteins are cell-specific transcription factors whose specificity of action has been attributed to protein-DNA and protein-protein interactions mediated by their DNA-binding (POU) domains. Here we report that transcriptional activation by SCIP, a POU protein expressed by developing Schwann cells, is dependent on an amino-terminal effector domain and that this domain mediates cell-specific transactivation in the complete absence of the POU domain. When fused to a heterologous DNA-binding domain, this SCIP domain is a potent transactivator in Schwann cells but is inactive in three heterologous cell types. The primary structure of the SCIP amino-terminal domain is novel but contains a polymorphic string of alanine residues similar to those found in several other transcription factors. Although previously hypothesized to be important for transcription factor activity, we find that the SCIP string is functionally irrelevant. We propose that homopolymers of alanine, and certain other amino acids, do not represent a motif required for transcription factor function but instead reflect regions of unstable DNA related to those associated with four recently characterized human genetic disorders. PMID- 8234345 TI - Characterization of composite aminodeoxyisochorismate synthase and aminodeoxyisochorismate lyase activities of anthranilate synthase. AB - Anthranilate synthase [chorismate pyruvatelyase (amino-accepting), E.C.4.1.3.27] catalyzes the formation of anthranilate (o-aminobenzoate) and pyruvic acid from chorismate and glutamine. A mutant form of the enzyme from Salmonella typhimurium accumulates a compound that we had isolated and identified as trans-6-amino-5-[(1 carboxyethenyl)-oxy]-1,3- cyclohexadiene-1-carboxylic acid, commonly called aminodeoxyisochorismate (ADIC). Here we report that ADIC is formed by a reversible, Mg(2+)-dependent ADIC synthase activity of anthranilate synthase that can be functionally uncoupled from a Mg(2+)-dependent ADIC lyase activity of the enzyme by single amino acid substitutions in the TrpE subunit of the anthranilate synthase complex of S. typhimurium. Both of the component activities of the enzyme are sensitive to feedback inhibition by L-tryptophan. Purified ADIC is quantitatively converted to anthranilate and pyruvic acid by the ADIC lyase activity of wild-type anthranilate synthase. ADIC also serves as a substrate for the formation of chorismate by the enzyme in the absence of glutamine and (NH4)2SO4. The rate of ADIC formation by the mutant enzyme and the steady-state parameters for ADIC utilization by the wild-type enzyme are consistent with a role for ADIC as an enzyme-bound intermediate that does not accumulate during the course of the anthranilate synthase reaction. The altered catalytic specificity of mutant anthranilate synthase enzymes suggests a potential role for ADIC in secondary metabolism. PMID- 8234346 TI - D-chiro-inositol metabolism in diabetes mellitus. AB - D-chiro-inositol is a rare inositol isomer present in inositol phosphoglycans which are proposed mediators of insulin action. To study D-chiro-inositol metabolism in diabetes mellitus, a sensitive and specific assay was developed using negative-ion chemical ionization gas chromatography/mass spectrometry. Median urinary D-chiro-inositol excretion, which was 2.1 mumol/day in nondiabetics, was substantially increased to 12 mumol/day in non-insulin dependent diabetes (P < 0.0001) and to 74 mumol/day in insulin-dependent diabetes (P < 0.0001). Urinary D-chiro-inositol was strongly correlated with fasting plasma glucose (r = 0.568, P < 0.0001), glycated hemoglobin (r = 0.529, P < 0.0001), and urinary glucose (r = 0.368, P = 0.01). The renal clearance of D chiro-inositol was selectively elevated in both non-insulin-dependent and insulin dependent diabetes when compared with the clearances of L-chiro-inositol or myo inositol and exceeded the glomerular filtration rate in 71% of the diabetics but in none of the nondiabetics. In poorly controlled diabetic patients insulin treatment reduced urinary D-chiro-inositol losses by 63% and increased plasma levels by 8.8-fold. The metabolism of D-chiro-inositol is abnormal in diabetes and appears to be influenced by short- and long-term metabolic control. PMID- 8234347 TI - Activation by nitric oxide of an oxidative-stress response that defends Escherichia coli against activated macrophages. AB - Nitric oxide is a free radical (NO) formed biologically through the oxidation of L-arginine by nitric oxide synthases. NO is produced transiently in mammalian cells for intercellular signaling and in copious quantities to cause cytostasis and cytotoxicity. In the latter situation, NO is a deliberate cytotoxic product of activated macrophages, along with other reactive oxygen species such as hydrogen peroxide (H2O2) and superoxide (O2-). Escherichia coli has a complex set of responses to H2O2 and O2- that involves approximately 80 inducible proteins; we wondered whether these bacteria might induce analogous defenses against nitric oxide. We show here that a multigene system controlled by the redox-sensitive transcriptional regulator SoxR is activated by NO in vivo. This induction confers bacterial resistance to activated murine macrophages with kinetics that parallel the production of NO by these cells. Elimination of specific SoxR-regulated genes diminishes the resistance of these bacteria to the cytotoxic macrophages. The required functions include manganese-containing superoxide dismutase, endonuclease IV (a DNA-repair enzyme for oxidative damage), and micF, an antisense regulator of the outer membrane porin OmpF. These results demonstrate that SoxR is a sensor for cellular exposure to NO, and that the soxRS response system may contribute to bacterial virulence. PMID- 8234348 TI - Characterization of functional organization within rat barrel cortex using intrinsic signal optical imaging through a thinned skull. AB - We used optical imaging of intrinsic signals to characterize the functional representations of mystacial vibrissae (whiskers) in rat somatosensory cortex. Stimulation of individual whiskers for 2 s at 5 Hz resulted in a discrete area of functional activity in the cortex. Images of whisker representations were collected both through the dura and through a thinned skull. We characterized the functional representation of a whisker both spatially and temporally with two dimensional images and three-dimensional surface plots of intrinsic signal development in the cortex in response to whisker stimulation. Single unit recordings verified that the representation of the whisker obtained with optical imaging corresponded with the electrophysiological response area of that whisker in the cortex. Lesions in the center of the functional activity were found to be in the center of the dense cytochrome oxidase patch for the corresponding whisker. In addition, a 3 x 3 matrix of whiskers was stimulated and the distances between the centers of the imaged representations and the distances between the centers of the layer IV cytochrome oxidase staining of the nine whiskers were found to be highly correlated (r = 0.98). This study shows a striking correspondence among imaging, physiology, and anatomy in the rat somatosensory cortex. Furthermore, the ability to use optical imaging through a thinned skull should allow investigations into the long-term changes in a sensory representation within a single animal. PMID- 8234349 TI - The Nestle Lecture. Chronic undernutrition and metabolic adaptation. PMID- 8234350 TI - Micronutrient deficiencies in the preterm neonate. AB - The nutrition of the premature infant poses a critically important challenge to clinicians. Premature infants are a heterogeneous group; maternal status, gestational age, drug intake, respiratory distress, phototherapy, and infection all conspire to make it extremely unlikely that a recommendation for daily intakes will satisfactorily encompass all babies. Clinical and subclinical deficiencies evidently do occur, and the impact of nutrient imbalance may have serious implications for outcome. If advances in clinical practice mean enhanced survival rates of babies of very small gestational age, then it is of vital importance that we work to establish the most appropriate regimens for vitamin and mineral intakes in this group. PMID- 8234351 TI - Genetic regulation of enterocyte differentiation. PMID- 8234352 TI - Molecular aspects of enzyme synthesis in the exocrine pancreas with emphasis on development and nutritional regulation. PMID- 8234353 TI - Dietary regulation of intestinal nutrient carriers. PMID- 8234354 TI - Influence of the weaning diet on the changes of glucose metabolism and of insulin sensitivity. PMID- 8234355 TI - Nutritional supply of proteins and absorption of their hydrolysis products: consequences on metabolism. PMID- 8234356 TI - Manipulation of the gut microflora: experimental approach in animals. PMID- 8234357 TI - Manipulation of the human gut microflora. PMID- 8234358 TI - Factors affecting fermentation reactions in the large bowel. PMID- 8234359 TI - Interactions between nutrition and the intestinal microflora. PMID- 8234360 TI - No reduction in the cost of mating for Drosophila melanogaster females mating with spermless males. AB - Drosophila melanogaster females can incur a cost of mating, manifested as a decrease in longevity and lifetime reproductive success. We investigated whether the cost of mating was a cost of storing and/or receiving sperm by using two types of males that do not transfer sperm (transformer pseudomales and the male offspring of homozygous tudor mothers). Females that were intermittently exposed to males that did and did not transfer sperm did not differ in lifespan, in the absence of any differences in other costly aspects of reproduction, showing that there was no cost to receiving sperm. There was a cost of mating with spermless males; this suggested a potential cost of receiving accessory fluid. However, it was not possible to distinguish this possibility from other explanations, e.g. female injury at mating and the transfer of parasites. The reasons why females continuously exposed to males remated more than was in their reproductive interests is discussed. PMID- 8234361 TI - Facilitation of sodium currents in frog neuroglia by nerve impulses: dependence on external calcium. AB - Facilitation of voltage-gated sodium currents in glial membranes by nerve impulses has been studied by using both the whole cell and loose-patch clamp techniques in the isolated intact optic nerve of the frog. During facilitation there is a shift in the voltage dependence of glial Na+ channels such that a given depolarization produces a larger inward Na+ current. Decreasing external calcium from 4 times normal to 0.2 times normal produced a similar shift in the current-voltage relation. Increasing the external calcium concentration to 4-5 times normal blocks facilitation. In reduced calcium, 0.1-0.2 times normal, the peak of facilitation was unaffected, but its decay was slowed. The addition of 1 mM nickel and 2 mM cobalt or 2 mM cadmium, to prevent depletion of extracellular calcium that might result from voltage-dependent entry of calcium into the axons, did not block the facilitation. The results suggest that, even though facilitation is blocked by high extracellular calcium, a decrease in extracellular calcium produced by axon impulses is not the cause of the facilitation. PMID- 8234362 TI - Detecting chaos in a noisy time series. AB - We propose a new method for detecting low-dimensional chaotic time series when there is dynamical noise present. The method identifies the sign of the largest Liapunov exponent and thus the presence or absence of chaos. It also shows when it is possible to assign a value to the exponent. This approach can work for short time series of only 500 points. We analyse several real time series including chickenpox and measles data from New York City. For model systems it correctly identifies important spatial scales at which noise and nonlinear effects are important. We propose a further technique for estimating the level of noise in real time series if it is difficult to detect by the former method. PMID- 8234363 TI - Sperm competition games: sperm size and sperm number under adult control. AB - Evolutionary games of sperm competition in which two males mate with the same female have previously considered sperm size to be fixed at some (small) constant level. Although male gametes in multicellular organisms are typically small compared with ova, they vary greatly both between and within groups, and sperm size sometimes correlates with the probability of sperm competition. This paper examines 'raffle principle' sperm competition games in which both size and number of gametes can be varied strategically under control of the diploid parent. If ejaculate investment trades off against the number of matings that a male can achieve, the evolutionary stable strategy (ESS) ejaculate expenditure (as a fraction of reproductive effort per mating) approximates to p/4 when the probability, p, of sperm competition is low. Sperm size may either: (i) increase a sperm's competitive weight (a measure of its advantage in the fertilization raffle), or (ii) influence its mortality rate in the female tract. On the simplest model, size is optimized after the marginal value theorem, and may be large or small depending on how size influences competitive weight or survivorship. Further, sperm size is independent of the risk of sperm competition, and only sperm numbers increase with this risk. However, some recent studies show sperm size to increase with sperm competition. The present analysis offers the following possibilities: (i) there are unidentified constraints on sperm number, so that ejaculate mass can increase only by increase in sperm size; (ii) competitive benefits of size become more important as sperm numbers increase; (iii) size mainly increases survivorship, and sperm competition risk increases with the mean duration between mating and fertilization; and (iv) size increases competitive ability at the expense of survivorship, and sperm competition risk decreases with time between mating and fertilization. These conclusions relate to advantages conferred by size on sperm before fertilization; they do not affect the prediction of previous models that no component of sperm size should evolve for provisioning the zygote. PMID- 8234364 TI - Sperm competition games: sperm size and number under gametic control. AB - This paper examines sperm competition games in which both size and number of gametes in an ejaculate can be varied strategically, but under the control of the haploid sperm. It is a companion to the previous paper (Parker (Proc. R. Soc. Lond. B 253, 245 (1993))) for the case where the strategic control is by the diploid parent. Under haploid control, the optimal balance between size and number of sperm in an ejaculate becomes quite different from that predicted for diploid control; there is a conflict between parental and gametic interests over sperm size and number. This occurs because both intra-ejaculate and inter ejaculate sperm competition are present with haploid control, whereas intra ejaculate competition is absent under diploid control. The magnitude of this conflict is affected by the risk of inter-ejaculate sperm competition, and by the fact that gametic interests depend on the nature of available mutations. If ejaculate expenditure trades off against numbers of matings achieved, with haploid control there is escalation towards maximal expenditure on the ejaculate. If the ejaculate expenditure is fixed by the diploid parent, but there is a size number trade-off under haploid control, solutions depend on the mode of action of the mutation affecting sperm size or number. If mutant-bearing sperm deviate entirely at the expense of non-mutant sperm, any increase in size or number will spread. If mutant sperm deviate entirely at their own expense, the evolutionarily stable strategy (ESS) is the same as for diploid control; there is no conflict.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234365 TI - Badgers (Meles meles), cattle and bovine tuberculosis (Mycobacterium bovis): a hypothesis to explain the influence of habitat on the risk of disease transmission in southwest England. AB - Badgers are believed to be responsible for a high proportion of the cases of bovine tuberculosis in cattle in southwest England where, despite the onset of badger control operations in 1975, comparatively high numbers of cattle continue to fail the tuberculin test. To determine why the disease remains a problem in these areas, data on badger densities and patterns of land use were examined. Areas subject to repeated badger control operations had greater landscape heterogeneity and a higher density of linear habitat features. These habitat features were not related to badger density, measured as the mean number of social groups per square kilometre. Environmental contamination by infected badger urine is thought to be the main mode for the transmission of bovine tuberculosis. Field studies in an area with tuberculosis in both badgers and cattle showed that badgers may urinate on pasture after crossing through a linear feature, and that the number of these crossing-point urinations increases with the number of linear features crossed. The hypothesis is presented that these crossing-point urinations are a major source of bovine tuberculosis infection in cattle, and that areas with increased numbers of linear features have greater levels of contamination of pasture with badger urine and hence increased opportunities for disease transmission. PMID- 8234366 TI - Accounting for bias in estimates of the rate of polygenic mutation. AB - Experimental data on the rate of response to artificial selection in initially inbred lines or the rate of divergence among inbred sublines can be used to estimate the rate of increase in variance of quantitative traits from new mutations. So far estimates have been based on the infinitesimal model of many genes with small additive effects which imply a rate of increase in heritability for Drosophila melanogaster bristle number traits of about 0.1% per generation. Such estimates are biased because mutants tend to have large effects, to have non additive gene action, and to be deleterious. Here, recent information on the distribution of effects of new mutations on Drosophila melanogaster bristle number and viability is used to infer the direction and magnitude of this bias. The infinitesimal model tends to underestimate the mutational variance, typically by a factor of about 3, but this factor depends on the experimental design. Averages of revised estimates, accounting for this bias, of the per generation increment in heritability from mutation are 0.36% and 0.21% for abdominal and sternopleural bristle number, respectively, in experiments involving M strains, and 1.4% and 0.7% for abdominals and sternopleurals, respectively, in P strains. PMID- 8234367 TI - Presynaptic inhibitory action of a metabotropic glutamate receptor agonist on excitatory transmission in visual cortical neurons. AB - A family of metabotropic glutamate receptors (mGluRs) has been elucidated by molecular cloning. To study the possible modulatory role of mGluRs in synaptic transmission, we tested the effect of a mGluR agonist, (+/-)-1-aminocyclopentane trans-1,3-dicarboxylic acid (trans-ACPD), on the excitatory post-synaptic currents (EPSCS) recorded from neurons in thin slices of rat visual cortex, by using the whole-cell patch-clamp method. We found that trans-ACPD markedly suppressed the evoked EPSCS without affecting the mean amplitude of spontaneous miniature EPSCS. This effect on the evoked EPSCS was blocked by a potassium channel blocker, 4-aminopyridine (4-AP) in a dose-dependent manner. We suggest that trans-ACPD presynaptically inhibits EPSCS by a mechanism involving the 4-AP sensitive channels. PMID- 8234368 TI - Society for Experimental Biology and Medicine. 1994. Constitution and bylaws. Membership directory. PMID- 8234369 TI - Hepatic progenitor populations in embryonic, neonatal, and adult liver. AB - Oval cells, small cells with oval-shaped nuclei, are induced to proliferate in the livers of animals treated with carcinogens and are thought to be related to liver stem cells and/or committed liver progenitor cell populations. We have developed protocols for identifying and isolating antigenically related cell populations present in normal tissues using monoclonal antibodies to oval cell antigens and fluorescence-activated cell sorting. We have isolated oval cell antigen-positive (OCAP) cells from embryonic, neonatal, and adult rat livers and have identified culture conditions permitting their growth in culture. The requirements for growth of the OCAP cells included substrata of type IV collagen mixed with laminin, basal medium with complex lipids and low calcium, specific growth factors (most potently, insulin-like growth factor II and granulocyte macrophage colony-stimulating factor), and co-cultures of embryonic, liver specific stroma, strongly suggesting paracrine signaling between hepatic and hemopoietic precursor cells. The growing OCAP cultures proved to be uniformly expressing oval cell markers but were nevertheless a mixture of hepatic and hemopoietic precursor cells. To separate the hepatic and hemopoietic subpopulations of OCAP cells, we surveyed known antibodies and found ones that uniquely identify either hepatic or hemopoietic cells. Several of these antibodies were used in panning procedures and fluorescence-activated cell sorting to eliminate contaminant cell populations, particularly hemopoietic and endothelial cells. Using specific flow cytometric parameters, three cellular subpopulations could be isolated separately that were identified by immunochemistry and molecular hybridization assays as probable: (i) committed progenitors to hepatocytes; (ii) committed progenitors to bile ducts; or (iii) a mixed population of hemopoietic cells that contained a small percentage of hepatic blasts that are possibly pluripotent. The hepatic precursor cells have been characterized using immunochemistry, flow cytometry, and molecular hybridization assays. The hepatic blasts are small (7-10 microns) cells with high nuclear to cytoplasmic ratios and with minimal complexity of the cytoplasm. Cultures of the committed progenitors were found to differentiate into cells with recognizable parenchymal cell fates. We discuss our studies in the context of our model of the liver as stem cell and lineage system and suggest that a slow, unidirectional, terminal differentiation process, paralleling more rapid ones in the skin or gut, occurs at all times in the liver and is thought to vary primarily in kinetics during quiescent versus regenerative states.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234370 TI - Isolation, culture, and transplantation of rat hepatocytic precursor (stem-like) cells. AB - From a review of past studies and the report of new studies from our laboratory, this article provides strong evidence to show that WB-F344 (WB) rat liver epithelial cells are stem-like precursor cells for hepatocytes. WB cells are structurally and phenotypically simple epithelial cells that were isolated from the liver of an adult male Fischer 344 rat, under conditions that excluded their origin from hepatocytes in vivo. WB cells express a phenotypic repertory that overlaps, but is distinct from, that of both hepatocytes and bile duct epithelial cells. The complex phenotype of WB cells is compatible with their being embryonic or undifferentiated variants of either hepatocytes or bile duct epithelial cells. When WB cells are tagged genetically with genes for bacterial beta-galactosidase and neomycin resistance (BAG2-WB), they and their progeny can be distinguished from parental WB cells and hepatocytes by the expression of these gene products. Progeny of BAG2-WB cells that were transplanted into the liver parenchyma of syngeneic rats integrated into hepatic plates and acquired the morphological and functional attributes of adjacent host hepatocytes; the progeny of BAG2-WB cells in the liver express albumin, tyrosine aminotransferase, alpha-1-antitrypsin, and transferrin. We also demonstrate that progeny of BAG2-WB cells can be recovered from livers into which they have been transplanted, which may allow the elucidation of alterations in gene expression that accompany their differentiation. PMID- 8234371 TI - Phenotypic heterogeneity within clonogenic ductal cell populations isolated from normal adult rat liver. AB - Oval cells represent a heterogeneous population composed of ductal cells, transitional cells with characteristics of both hepatocytes and bile ductal cells, and bipotential stem cells capable of differentiation along a biliary or hepatocytic lineage. In an attempt to define markers that would distinguish between individual cell types within the oval cell population, a number of investigators have utilized hybridoma technology to produce cell type-specific monoclonal antibodies. Several of these have proved to be of value in delineating lineage relationships during fetal development and carcinogenesis in the adult liver. Most recently, monoclonal antibodies specific for OC2 and OC3, two oval cell antigens identified in our laboratory, have been used in combination with magnetic beads or a fluorescence-activated cell sorter to isolate antigenically defined subpopulations from adult and fetal rat liver. Using OC2-positive fetal liver cells as an immunogen, we have produced a monoclonal antibody identifying a bile ductal antigen, designated BD1, that is differentially expressed by oval cells and normal ductal cells. This antigen shows a heterogeneous pattern of reactivity that defines three distinct cell populations in regenerating rat liver: a BD1-negative, [3H]thymidine-labeled cell population thought to contain hepatic stem cells; a BD1-positive, thymidine-negative population of terminally differentiated ductal cells; and a BD1-positive, [3H]thymidine-positive population of mature ductal cells. Analysis of BD1 expression in vitro on continuous lines of bile duct epithelial cells (BDEC) demonstrated that BD1 was rapidly increased in late G1 and lost during G2/M. High passage cultures of BDEC and primary cultures of oval cells expressed low or undetectable levels of BD1 and high passage BDEC failed to express BD1 when arrested in late G1. Taken together, these results suggested that oval cells and high passage BDEC might share a subtle defect in cell cycle regulation marked by an inability to upregulate the expression of BD1. PMID- 8234372 TI - Gene therapy for human hemoglobinopathies. AB - Gene transfer of human globin genes into human pluripotent stem cells via viral vectors may soon be realized. The high level of globin gene expression believed to be required for the treatment of severe hemoglobinopathies necessitated the inclusion of cis-acting sequences (LCR). Retroviral vectors containing the LCR elements are prone to rearrangement, low titer, and poor expression. Inclusion of a "minilocus" containing four HS sites linked to a globin gene resulted in higher expression in transplanted mice, but rearrangement of the provirus still occurs, and it is unclear what significance these experiments have with regard to human marrow stem cell transduction. Recombinant AAV is among the newest of genetic transfer vectors. This once obscure virus possesses unique properties that distinguish it from all other vectors. Its major advantage is the lack of pathogenicity in humans. Wild-type AAV has the unusual ability to selectively integrate into the mammalian genome at a specific region, thus reducing the concern for genomic disruption and insertional mutagenesis. The ability of AAV to carry regulatory elements without interference from the viral template may enable greater control of transferred gene expression. Disadvantages currently include the inferior packaging systems which yield low numbers of recombinant virions which are contaminated with wild-type adenovirus. The small AAV genome that can be packaged (approximately 5 kb) rules out its use for transfer of larger genes. Recombinant AAV viruses do not appear to demonstrate the same site-specific genomic integration as wild-type viruses. Elucidation of the mechanism of site specific integration should prove useful in the development of safe vectors for gene transfer as well as provide insight into the nature of DNA recombination in humans. PMID- 8234373 TI - Molecular cloning and characterization of a cDNA encoding feline interleukin-6. AB - With the discovery that the cat is susceptible to infection by a lentivirus quite similar to the virus that causes the human acquired immune deficiency syndrome, the cat has become an important animal model with which the interactions between host and immunodeficiency virus can be studied. To facilitate investigation of the feline immune response and the response of cytokine expression to infection by feline immunodeficiency virus, our laboratory has focused on the isolation and molecular cloning of cDNA representative of feline lymphokines, cytokines, and monokines. Herein, we describe the molecular cloning and characterization of a cDNA encoding feline interleukin (IL)-6. The nucleotide sequence of feline IL-6 was found to exhibit 81%, 76%, 63%, and 61% homology with pig, human, rat, and mouse IL-6, respectively, while the predicted amino acid sequence exhibits 66%, 53%, 37%, and 30% homology with pig, human, rat, and mouse IL-6, respectively. PMID- 8234374 TI - Antitumor activity of an immobilized peroxidase system against murine Ehrlich ascites is mediated by the immune system. AB - Earlier studies have shown that a mixture of glucose oxidase and a peroxidase exerts a tumoricidal effect on rats bearing Novikoff hepatomas when the enzyme mixture is injected intraperitoneally. The enzyme mixture was shown to be nontoxic when injected into healthy animals at levels up to 600 times the therapeutic dose. In the present study, we have evaluated the possibility that the host immune defense system may be involved in the antitumor activity of the peroxidase system, using the murine Ehrlich ascites tumor as the target. The results revealed that the antitumor activity of the peroxidase system is absent in tumor-bearing animals whose immune system has been compromised by whole body gamma-irradiation or by an induced selenium deficiency. The peroxidase system was also found to be inactive in tumor-bearing mice whose immune system was suppressed by the administration of cyclosporin A as well as in athymic (nu/nu) mice. These results indicate that T lymphocytes may directly or indirectly be involved in the in vivo antitumor activity of the peroxidase system. This could explain the observed high selectivity toward tumor cells by the enzyme system in vivo and its lack of toxicity in healthy animals. PMID- 8234375 TI - Fibronectin synthesis by aorta explants from rabbits fed high cholesterol diets. AB - Fibronectin synthesis was studied in aorta explants in culture from rabbits fed a high fat-high cholesterol diet. [3H]Mannose and [14C]leucine were used to label oligosaccharide side chains and the protein core, respectively. The synthesis was followed by monitoring immunoprecipitable fibronectin from the culture medium using polyclonal goat anti-rabbit fibronectin antibody. Synthesis of fibronectin increased by [14C]leucine (81%) and [3H]mannose (29%) incorporation over controls. On gel filtration, fibronectin synthesized by controls and cholesterol fed rabbit resolved into four fractions. Pulse-chase experiment with [3H]glucosamine or [3H]leucine showed that fibronectin secreted by the aorta explants from rabbits fed high fat-high cholesterol diets incorporated an increased amount of radioactivity. Pulsing with [3H]mannose showed decreased incorporation of the label. During the chase period, the rate of secretion of fibronectin into the media by the hypercholesterolemic rabbit aorta explants was increased. The fibronectin that bound to the gelatin or heparin columns from cholesterol-fed rabbit aorta media had lower levels of [3H]mannose incorporated into the glycoprotein than the control. These results indicate that there is an alteration in carbohydrate composition of the fibronectin synthesized by the aorta explants from rabbits fed a high cholesterol diet. High fat-cholesterol intake could play a causative role in matrix dysfunction during atherogenesis by altering glycoprotein synthesis. PMID- 8234376 TI - Open discussion can lead to acceptance. The psychosocial effects of stoma surgery. AB - 1. Psychological and physical healing do not necessarily follow the same timepath. 2. Patients should be allowed to talk through their feelings. 3. Stoma surgery involves a profound sense of loss. 4. Patients should rehearse their responses with others. PMID- 8234377 TI - Living with a stoma. PMID- 8234378 TI - A tool to fill the gap. Developing a wound risk assessment chart for children. AB - 1. Consistent monitoring and assessment of wounds is vital in children as well as adults. 2. There are anatomic, physiological and psychological differences between adults and children, of which nurses should be aware. 3. To date, the practice of paediatric risk assessment is an unknown entity. 4. A pressure sore risk assessment chart was developed specifically for use with children. PMID- 8234380 TI - Community-versus hospital-based services for psychiatric care. PMID- 8234379 TI - Informed choices for long-term benefits. The management of catheters in continence care. AB - 1. Non-invasive methods of managing urinary dysfunction should be considered as alternatives to long-term catheterisation and reviewed on a regular basis. 2. Nurses must match choice of catheter and drainage equipment with the individual patient, to enhance self-care and independence. 3. Monitoring of catheter life can establish a pattern of recurrent blockage and allow pro-active, planned recatheterisation. 4. Good handwashing practice is important in catheter management to reduce the risk of cross-infection. PMID- 8234381 TI - Fulfilling the commitment to improve patient care? Assessment of the effect of the Patient's Charter in a community hospital. AB - 1. Interviewing patients provides important information on the priorities they place on their care. 2. Patient-centred standards based on this information can then be set by multidisciplinary groups. 3. Auditing these standards can lead to improved patient care. 4. Audit should be regarded by professionals as a positive process. PMID- 8234382 TI - The multiple benefits of accurate assessment. Effective management of leg ulcers. AB - 1. To be effective, the treatment of leg ulcers should be directed at the underlying cause. 2. If the underlying cause is chronic venous insufficiency, the most effective treatment is compression therapy. 3. There are a number of effective bandaging systems in use; the choice of any particular regimen is of less importance than the skill of the practitioner in applying it. 4. The prevention of recurrence should form a significant part of the care plan. PMID- 8234383 TI - The Norton scale revisited. PMID- 8234384 TI - Will the government take the long-term view? PMID- 8234385 TI - A figurehead that leads by example. Effective leadership through the management of resources. AB - A figurehead that leads by example: effective leadership through the management of resources. Professional Nurse, 9, 2, 80-84. 1. It is essential that encouragement and praise are given where appropriate. 2. It is important to support teams, even if they fail. 3. Managers need to be flexible in their thoughts and actions. 4. Managers are also required to be vigilant to what is happening around them and to the needs of the team. PMID- 8234386 TI - Let the team decide? Evaluation of self-rostering on an acute general medical ward. AB - 1. Self-rostering enables staff to develop their managerial skills. 2. Self rostering combined with a system of team nursing could allow the team to function at a high level of self-reliance and autonomy. 3. This project indicated that these outcomes could be achieved, but as with any implementation of change, the time required for staff preparation should never be underestimated. PMID- 8234387 TI - Managing chemotherapy-induced nausea and vomiting. AB - 1. Nausea and vomiting can further debilitate patients in whom physical and psychological status is already compromised, and may seriously diminish their quality of life. 2. A number of nursing, psychological and pharmacological measures can be employed to reduce these side-effects. 3. While conventional anti emetics have afforded some control of emesis, the 5-HT3 antagonists have proved to be effective and well-tolerated agents. PMID- 8234388 TI - A guide to advanced trauma life support. AB - 1. ATLS is a systematic approach, aimed at reducing mortality in the 'golden hour' after injury. 2. Injury to the cervical spine must be presumed, and effective immobilisation methods employed in all trauma patients until such an injury can be medically excluded. 3. Blood loss is the most common remediable cause of morbidity and mortality following trauma. 4. Many A&E nurses lack confidence in care of trauma patients and require further training in this area. PMID- 8234389 TI - Practical skills to overcome emotional problems. Developing a model of cognitive behavioural therapy nursing for acute inpatient psychiatric care. AB - 1. Cognitive behavioural therapy is increasingly being found helpful in treating various psychiatric disorders. 2. A nursing model is proposed that incorporates this therapy into inpatient psychiatric care. 3. The model may be helpful in panic, agoraphobia, self-harm and psychosis, and has the attraction that progress is measurable. PMID- 8234390 TI - Encapsulation of drugs and excipients in liposomes--measurements with drug specific electrodes. AB - In this study, the degree of encapsulation of benzalkonium chloride in liposomes was quantitatively measured using a potentiometric membrane electrode specific for benzalkonium chloride. The encapsulation of lidocaine hydrochloride was examined with another ion-selective electrode for comparison. Liposomes were prepared from a commercially available liposome concentrate (Phosal 75 SA). Photon correlation spectroscopy was used to detect the formation of liposomes in the size range of 200 nm. The measurements with the membrane electrode enabled the activity of the free drug to be quantitatively determined in the presence of liposomes. The investigations showed that, in the concentration range examined, up to 97% of the amphiphilic benzalkonium chloride is encapsulated in the liposomes. In the case of the hydrophilic lidocaine hydrochloride, virtually no liposomal encapsulation occurs. PMID- 8234391 TI - Stability of some phenolic antioxidants in fatty preparations. AB - Stability of butylated hydroxyanisole (BHA), propyl gallate (PG), trihydroxy butyrophenone (THBP), tert-butyl hydroquinone (TBHQ) and a mixture of PG and BHA (1:1) in fatty preparations stored under three different conditions was studied. Each antioxidant was added to a fatty model to determine its degradation kinetics and its evolution during the oxidation of the fatty product. Their degradation was adequately described by a first-order kinetic process. An opposite relationship between the stability of antioxidants and their rate protection against oxidation of the fatty product was obtained. At 25 degrees C, t90 values ranged between 10 days for TBHQ and 61 days for BHA. On the other hand, compared when they were formulated alone, the increase of PG and the diminution of BHA degradation kinetics were observed. A degradation product arising from TBHQ was found in samples and stock solutions; an HPLC method for its identification is proposed with detection at 254 nm. PMID- 8234392 TI - Adenosine receptors and their modulators. AB - The identification and characterization of adenosine receptors and the development of potent, receptor subtype-selective agonists and antagonists has been an active area of research for the past 20 years. Major recent advances in the field have been the cloning of several adenosine receptor subtypes of different species, including the discovery of a new subtype, designated A3, the discovery and development of new agonists and antagonists, particularly those with selectivity for the A2a adenosine receptor, the characterization of signal transduction pathways, and the development of agents which act indirectly on the adenosine receptor system. The present article focusses on aspects of pharmaceutical/medicinal chemistry related to adenosine receptors. PMID- 8234393 TI - [Barbiturate analysis with heavy metal ions from the beginning to the present]. PMID- 8234394 TI - [Computers in pharmacy--an overview]. PMID- 8234395 TI - [Process control and automatization of high-speed tablet presses]. PMID- 8234396 TI - [Xyloglucan--structure, genesis, and functions of a widely distributed substance group]. PMID- 8234397 TI - The concept of fractals in the pharmaceutical sciences. AB - An old problem is sometimes better understood when it is viewed from a new perspective than by investigating it repeatedly with classical methods. Such a new perspective is provided by the concept of fractals. Briefly, this term defines a spread class of geometric shapes, whose subunits replicate the structure of the larger unit in accordance with the formalization of the idea of self-similarity. This concept was introduced by Mandel-brot in respond to the need for a more sophisticated explanation of numerous phenomena commonly encountered in nature. In this review we will provide a short introduction into the concept of fractals and a concise overview of those studies where fractal geometry has been employed so far in pharmaceutical research. We believe that a review of the most recent findings in the application of fractal geometry to problems encountered in the pharmaceutical sciences might be helpful also to those people who are involved in the search for novel drug substances, as well as in the investigation of their performance in vitro and in vivo. A few suggestions will be made where this concept probably could be helpful in the future and instructions will be given on how the fractal dimension of rugged objects can be estimated practically. PMID- 8234398 TI - Recent evaluations of thiosemicarbazones and semicarbazones and related compounds for antineoplastic and anticonvulsant activities. AB - The antineoplastic activities of various carbocyclic, acylic and heterocyclic thiosemicarbazones as well as a number of semicarbazones are outlined. In addition, the anticonvulsant properties of a variety of cyclic and acyclic thiosemicarbazones and semicarbazones are discussed. PMID- 8234399 TI - [Synthesis of 4-methyl-6-phenyl-thieno(2,3-d)pyrimidines with a formamidino- or oxalamidocarbonic acid residue with antianaphylactic activity]. AB - 3-Amino-4-methyl-6-phenyl-thieno[2,3-d]pyrimidine-2-carbonic acid alkylesters 1a, b were hydrolyzed to the potassium salt of the carbonic acid 2. Cyclization of 2 with acetanhydride yielded the tricyclic 1,3-oxazinone derivative 4. This compound reacted with pyrrolidine by different conditions of reaction to give the bisamide 7, the acetamidino carbonic acid 5 and their decarboxylated product 6. Compound 1a yielded with Vilsmeier reagents the formamidino compound 3. 3-Amino thieno[2,3-d]pyrimidin-2-carbonitrile gave under different conditions of reaction with oxalic acid diethylester or with oxalic acid ethylester chloride the tetracyclic 4-methoxy-9-methyl-7-phenyl-thieno[2,3-d:4,5-d']dipyrimidine-2-car bonic acid methylester 10 or the N-(2-cyano-4-methyl-6-phenyl-thieno[2,3 d]pyrimid-3-yl)oxalamid ic ethylester 12. These compounds were hydrolyzed to give the carbonic acids 11 and 13. Some of the synthesized substances showed an antianaphylactic activity. PMID- 8234400 TI - [Determination of aluminum traces in dialysis concentrates]. AB - To determine traces of aluminium in dialyse concentrates trouble free and accurate by atomic absorption spectrometry a special procedure of enrichment was developed. The aluminium chromazurol S complex is adsorbed selectively at polyethylene powder whereby the disturbanced matrix (KCl) is separated. After elution by a 0.3% ethanolic solution of nitric acid the determination of aluminium is carried out by the graphite oven (flameless) atomic absorption spectrometry. All used materials have to be cleaned carefully. The determination limit for aluminium is 5 micrograms/l dialyse concentrate (the IUPAC commended limit is 15 micrograms/l). PMID- 8234401 TI - Herbal mixtures with claimed slimming activity: determination by TLC and HPLC of illegally added drugs. AB - A method for the detection and quantitation of several undeclared drugs in herbal preparations with slimming activity is proposed. Samples containing various anorexics, hypoglycemics and antidepressants were prepared by addition of the drugs to a synthetic mixture containing the most commonly used plant powders for those preparations. Each sample was subjected to a treatment that permitted, after a simple ethanolic extraction, the identification of the drugs by TLC using three different solvent systems. A further purification of the ethanolic solution through a polyamide column allowed for quantitative analysis of the drugs by a RP HPLC method. The analytical recovery was good (88-97%); the calibration curves were linear over a wide range of drug concentrations (30-500 micrograms/ml) (r > 0.9995); the precision was high (CV% = 0.4-2.8) as well as the accuracy (96 102%). PMID- 8234402 TI - [Improvement of the absorption of the active substance 2'-hydroxy-5' methyllaurophenoxamine (FLM 5011) with prodrugs and preparations]. AB - Methods to improve the absorption of problem drugs are presented for the strongly lipophilic, poorly water-soluble, potential lipoxygenase inhibitor FLM 5011 (1). The water-solubility is improved by using prodrugs or by solubilizing. The characterization of solubility has been characterized with an suitable in vitro model system. The bioavailability of 1 in rabbits after oral administration is markedly increased using 1-prodrugs studied. The good correlation between the ABC measured in vitro and the AUC estimated in vivo demonstrates that it is possible to predict the bioavailability at the rabbit of highly lipophilic drugs such as 1 using the flow through model system. PMID- 8234403 TI - An in vitro percutaneous absorption study of non-ionic compounds across human skin. AB - Generally, the mechanisms of percutaneous absorption are studied from correlations between representative penetration parameters (permeability coefficients) and variables accounting for lipophilicity or other related physicochemical properties. The present study was developed on the basis of the in vitro permeability coefficients through human skin, 280 microns in thickness, of a non-ionic homologous series of compounds (phenylalkanols). The corresponding penetration/lipophilicity correlations were compared with those found for a basic homologous series (4-n-alkylanilines) through a membrane of similar characteristics. The in vitro behaviour of both series of compounds may be regarded as similar. Differences were only observed in the permeability coefficients of those elements which in both series exhibited lipophilicity values above that considered optimum, as predicted by the selected biophysical penetration model. Consequently, it appears that the removal of the thick dermal tissue leads to increased permeation rates for the more lipophilic compounds and suggests that the dermis acts as an aqueous matrix. PMID- 8234404 TI - Influence of 3-cyano-2-morpholino-5-(payrid-4-yl)pyridine (AWD 122-14) and dopamine on left ventricular function during acute volume load in 18-month-old spontaneously hypertensive rats and Wistar-Kyoto rats. AB - Effects of AWD 122-14, a new cardiotonic agent, and dopamine were studied in an experimental model of congestive heart failure in 12- and 18-month-old spontaneously hypertensive rats (SHR rats) in comparison to normotensive Wistar Kyoto rats (WKY rats) as control group. This model combines an acute volume overloading with an already existing chronic pressure overload. Heart rate (HR), peak left ventricular pressure (PLVP), left ventricular enddiastolic pressure (LVEDP), and left ventricular contractility index (LV dp/dtmax) were significantly elevated in SHR rats versus WKY rats. Left ventricular mass (LVM) to body mass (bw) ratio was increased in SHR rats and there was a parallel, rightward shift of the left ventricular diastolic pressure-volume-relationship in the 18-month-old SHR rats. Thus for a given LVEDP, there is an increased left ventricular enddiastolic volume (LVEDV) in SHR rats, indicating a true structural outgrowth of the left ventricular lumen. During acute volume overloading LVEDP increased in the 12- and 18-month-old WKY rats and in the 12-month-old SHR rats. In contrast, the 18-month-old SHR rats showed no increase of the already very high baseline level of LVEDP. Dopamine and AWD 122-14 increased LV dp/dtmax in all groups. AWD 122-14 was able to reduce left ventricular filling pressure of the 18-month-old SHR rats. A further interesting finding was that AWD 122-14 reduced the content of thiobarbituric acid material in the left ventricle in the 18-month-old SHR rats (reduced lipid peroxidation), suggesting a possible cardioprotective action of this substance. PMID- 8234405 TI - Hemodynamic response and effects on myocardial energetics of 3-cyano-2-morpholino 5-(pyrid-4-yl)pyridine (AWD 122-14) in anesthetized minipigs. AB - AWD 122-14, a new positive inotropic and vasodilating agent, was investigated in comparison to amrinone, milrinone and dopamine in anesthetized minipigs. AWD 122 14 (1.17.10(-7)-37.5.10(-7) mol/kg) increased dose-dependent left ventricular contractility (LV dp/dtmax) (122x5 +/- 11x3%; ED50 = 8.1x10(-7) to mol/kg). Dopamine (2.64x10(-8)-21x12 x 10(-8) mol/kg) in comparison increased contractility up to 153.1 +/- 44.9% of control value and is about 20 times more potent than AWD 122-14 at the ED50 value and about 10 times more potent at the ED30 value. Amrinone (1.60x10(-6)-16.90x10(-6) mol/kg) and milrinone (1.48x10(-7) 23.70x10(-7) mol/kg) only slightly increased contractility in anesthetized minipigs, but they appear to posses a similar pharmacological profile like AWD 122-14. The hemodynamic effects were associated with an increase in myocardial oxygen consumption (E1: 18.8 +/- 10.0%) due to the marked increases in LV dp/dtmax and heart rate. LVMW was unchanged and LVSW decreased (-29.0 +/- 10.2%) after application of AWD 122-14. The reduction in left ventricular work (LVMW, LVSW) and the increase in myocardial oxygen consumption led to a decrease of left ventricular external mechanical efficiency of the non-failing minipig heart (Etam: -21.1 +/- 9.4%). Additional hemodynamic effects of AWD 122-14 were studied under calcium channel blockade (verapamil, nifedipine). After pretreatment with verapamil the agent (1.17.10(-7)-18.75.10(-7) mol/kg i.v.) increased left ventricular contractility between 42.9 +/- 41.6% and 58.5 +/- 33.3%. After pretreatment with nifedipine the agent induced a dose-dependent increase in LV dp/dtmax between 11.1 +/- 7.7% and 47.8 +/- 23.7%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234406 TI - [Solid state sample preparation of ifosfamide and dechloroethyl- metabolites in biological material. 2. Consideration of the separation mechanism]. PMID- 8234407 TI - Transdermal delivery of drugs. Part 7: Evaluation of the adhesive layer of matrices for TDS. PMID- 8234408 TI - Pharmacological evaluation of certain potential novel local anesthetics. PMID- 8234409 TI - Effects of piracetam on N-methyl-D-aspartate receptor properties in the aged mouse brain. AB - Subchronic treatment of aged mice with piracetam (500 mg/kg p.o. for 14 days) elevates N-methyl-D-aspartate (NMDA) receptor density by about 20% and normalizes the enhanced affinity of L-glutamate for the NMDA receptor. Since deficits at the level of the NMDA receptor might be one of the mechanisms underlying age associated cognitive impairment, the effects reported for piracetam may be relevant for the cognition-enhancing properties of this drug. PMID- 8234410 TI - Interaction of tacrine at M1 and M2 cholinoceptors in guinea pig brain. AB - Tacrine (THA) selectively modulates binding of M1 ligands in an allosteric fashion causing positive cooperativity. The binding affinity of THA to M1 and M2 cholinoceptors is similar. It is therefore proposed that the allosteric selectivity of THA is a function of the binding site and not of THA itself. Its interaction of M1 and M2 cholinoceptors was examined in guinea pig brain homogenates using the selective M1 and M2 antagonists [3H]-pirenzepine ([3H]PZ) and [3H]AF-DX 384. The dissociation constants were 0.36 nmol/l for the M1 receptor and 0.23 nmol/l for the M2 receptor. We also compared the binding of THA and methoctramine (MTA) at M2 receptors. Tacrine displayed similar binding affinity for both M1 and M2 receptor subtypes. MTA was 100 times more potent an inhibitor of [3H]AF-DX 384 binding at M2 receptors than THA. In addition, THA was found to slow the dissociation of [3H]PZ from the M1 receptor. In contrast, the dissociation of [3H]AF-DX 384 from M2 receptor subtypes was unaffected. We conclude that THA acts as an agonist at M1 cholinoceptors because it slowed the dissociation of [3H]PZ. At M2 cholinoceptors its nature is that of an antagonist because it had no effect on [3H]AF-DX 384 dissociation. PMID- 8234411 TI - The effect of an opiate receptor antagonist on the ileal brake mechanism in the rat. AB - Studies investigated the effect of the opiate antagonist naloxone (10 mg/kg) on stomach to caecum transit (SCTT) during ileal infusion of saline or Intralipid. SCTT of the head of the meal was measured by hydrogen analysis and meal distribution by the radiolabelled meal technique. Intralipid delayed SCTT by delaying both gastric emptying (p < 0.01) and small bowel transit. Naloxone did not affect SCTT during ileal saline infusion, but produced a distal shift (p < 0.05) in the geometric centre of the meal and increased radioactivity in the caecum (p < 0.001) 100 min after gavage. Naloxone abolished the delayed SCTT of the meal induced by ileal lipid infusion, with an associated increase in radioactivity in the caecum at 200 min (p < 0.01), although the geometric centre was shifted proximally within the intestine (p < 0.01). The results suggest that the ileal brake is mediated in part by endogenous opiate pathways. PMID- 8234412 TI - Effect of chronic exposure to cold on vascular responsiveness to phenylephrine and angiotensin II. AB - Chronic exposure of rats to cold (5 degrees C, 3-4 weeks) results in the development of hypertension. To assess potential mechanisms by which this may occur, the vascular responsiveness to administration of phenylephrine (an alpha adrenergic agonist) and angiotensin II (AII) was studied in unanesthetized rats at 1, 3 and 5 weeks of exposure to cold (5 degrees C). Vascular responsiveness to intravenous administration of graded doses of phenylephrine was reduced in cold treated rats, the earliest effect being observed within 1 week of exposure. With respect to AII, vascular responsiveness to graded intravenous doses increased maximally within 1 week of exposure to cold and returned toward the level of the control group at 3 weeks. After 5 weeks of exposure to cold, it had returned to the level of the control group. These results suggest that vascular responsiveness to alpha-adrenergic stimulation appears to be directed toward prevention of an elevation of blood pressure in cold-treated rats. In contrast, vascular responsiveness to administration of AII is increased during the first 3 weeks of exposure to cold, at a time when plasma renin activity is also increased, and may thus play an important role in the initiation of cold-induced elevation of blood pressure. PMID- 8234413 TI - Effect of ryanodine on the contractile response of the normal and hypertrophied rabbit urinary bladder to field stimulation. AB - Bladder contraction, similar to most smooth muscle contraction, is dependent on both the translocation of extracellular calcium across the cell membrane, and the calcium-stimulated release of intracellular bound calcium. Partial outlet obstruction of the urinary bladder induces a marked increase in bladder mass which results in part from a substantial hypertrophy of the smooth muscle elements. In addition, the increase in bladder mass is associated with specific contractile dysfunctions. It is not known if the contractile dysfunction induced by partial outlet obstruction is related to alterations in calcium metabolism. Ryanodine is a pharmacological tool which can be utilized to study the role of intracellular calcium in mediating contractile events. Ryanodine stimulates the loss of intracellular calcium (bound in the sarcoplasmic reticulum) and reduces the participation of intracellular calcium in the contractile response to specific forms of stimulation. The current study investigates the effect of partial outlet obstruction on the ryanodine inhibition of the contractile response of rabbit urinary bladder to field stimulation. New Zealand White rabbits were anesthetized and catheterized with an 8-french Foley catheter. A 00 silk suture was surgically placed around the catheterized urethra, the incision closed, and the catheter removed. After 1, 3, 5 and 7 days of partial outlet obstruction, the bladder was rapidly removed and utilized immediately for contractile studies. The effect of ryanodine (0-80 microM) on the peak response to 2-, 8- and 16-Hz field stimulation at 0.6, 1.8 and 5.4 mM calcium was determined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234414 TI - Histomorphological studies on the effect of recombinant human superoxide dismutase in biochemically induced osteoarthritis. AB - Enzymatic scavenging of .O-2 radicals by injections of superoxide dismutase has been described to inhibit the free radical reactions resulting in tissue damage. Using a biochemically induced model of osteoarthritis (OA) in the knee joints of hens, we investigated the histomorphological alterations under therapy with recombinant human superoxide dismutase (rH-SOD) in various doses by histological histochemical grading. Treatment of experimental OA with rH-SOD (0.1 mg/0.1 ml intra-articularly) led to a significant reduction in the intensity of cartilage degradation. The pathomorphological alterations in the osteoarthritic cartilage of hens treated with rH-SOD were quantitatively but not qualitatively different from the placebo-treated group. Our results indicate that rH-SOD exerts an inhibitory effect on the deleterious processes on articular cartilage tissue during the course of OA and may counteract cartilage degradations induced or accelerated by oxygen radicals. PMID- 8234415 TI - The effect of endotoxin on tobramycin pharmacokinetics in young and aged rats. AB - The pharmacokinetics of tobramycin as a single intravenous dose (3 mg/kg) were determined in young (2-3 months) and aged (22-24 months) rats and in similar groups 24 h after a single intraperitoneal injection of endotoxin (5 mg/kg). In aged rats the systemic clearance of tobramycin was significantly lower than in young rats. Endotoxin significantly decreased tobramycin clearances in both age groups, the effect being more pronounced in aged rats. The endogenous creatinine clearance followed a similar pattern as the tobramycin clearance. The volume of distribution at steady state (Vdss) was significantly lower in aged rates than in young rats. Endotoxin tended to increase Vdss in both age groups. In aged rats, however, Vdss was still significantly lower than values from young rats. The mean residence time (MRT) and terminal half-life (t1/2 beta) of tobramycin were similar in young and aged rats. Endotoxin had no effect on these parameters in young rats when compared with their controls. In aged rats, however, endotoxin significantly prolonged both MRT and t1/2 beta when compared with normal aged rats and with young endotoxin-treated rats. PMID- 8234416 TI - Alprazolam metabolism in vitro: studies of human, monkey, mouse, and rat liver microsomes. AB - Biotransformation of the triazolobenzodiazepine alprazolam (ALP) was studied in vitro using hepatic microsomal preparations from human, monkey, mouse, and rat liver tissue. Two principal hydroxylated metabolites were identified: 4-hydroxy- and alpha-hydroxy-alprazolam (4-OH-ALP and alpha-OH-ALP). In all species, rates of 4-OH-ALP formation exceeded those of alpha-OH-ALP. In human liver microsomes, ratios of 4-OH-ALP/alpha-OH-ALP reaction velocities calculated at clinically relevant plasma concentrations of ALP ranged from 7 to 17, qualitatively consistent with, but numerically larger than, the ratio of the plasma levels of the two metabolites during clinical use of ALP in humans. Km values for both 4-OH ALP (170-305 microM) and alpha-OH-ALP (63-441 microM) considerably exceeded the usual maximum plasma concentration observed in humans (200 ng/ml, 0.65 microM), consistent with the linear (dose-independent) pharmacokinetic characteristics of ALP observed in humans. Thus formation of 4-OH-ALP via hydroxylation is the major route of ALP metabolism. This pathway is probably mediated by the cytochrome P 450-3A subfamily. Factors that impair the activity of this cytochrome subtype are likely to impair clearance of ALP in vivo. PMID- 8234417 TI - Pharmacokinetic-metabolic studies with 14C-aloe emodin after oral administration to male and female rats. AB - After oral administration of 4.5 mg/kg 14C-aloe emodin (AE) to rats 20-30% of the dose was excreted in urine and the rest in feces. AE was quickly metabolized to rhein, to an unknown metabolite and to conjugates of all three. In the plasma about 10% of 14C-activity was identified as free AE. Maximum plasma values were reached 1.5-3 h p.a. with 248 (male) and 441 (female) ng equivalents AE/ml. Maximum concentrations in plasma were about 3 times higher than those in ovaries and 10 times higher than those in testes. Only liver, kidney and intestinal tract showed higher concentrations than plasma. Terminal half-life (for radioactivity) in blood was about 50 h. PMID- 8234418 TI - Rhein and aloe-emodin kinetics from senna laxatives in man. AB - Therapeutic doses of two laxatives (Agiolax and Sennatin) were repeatedly administered to 10 healthy volunteers in a two-way change-over design. Blood samples were collected up to 96 h after the first dose, and plasma levels of total aloe-emodin and rhein were determined simultaneously with a sensitive (lower limit of quantification: 0.5 ng aloe-emodin and 2.5 ng rhein per millilitre plasma) and specific fluorometric HPLC method. Aloe-emodin was not detectable in any plasma sample of any subject. Rhein concentration time courses showed highest levels of 150-160 ng/ml and peak maxima at 3-5 h and 10-11 h after dosing probably according to absorption of free rhein and rhein released from prodrugs (e.g. sennosides) by bacterial metabolism, respectively. PMID- 8234419 TI - Cleavages of the O- and C-glucosyl bonds of anthrone and 10,10'-bianthrone derivatives by human intestinal bacteria. AB - A strictly anaerobic bacterium, Bifidobacterium sp. SEN, capable of hydrolyzing the O-glucosyl of sennosides was isolated from human feces. The bacterium stepwisely hydrolyzed sennoside B to sennidin B through sennidin-8-monoglucoside in PYF medium but not in GAM broth. Addition of D-glucose to PYF medium resulted in loss of the hydrolyzing activity in culture but addition of D-fructose did not affect the activity. Coculture of this bacterium with Peptostreptococcus intermedius led to rapid accumulation of rhein anthrone in the medium. Similarly, a bacterium, Eubacterium sp. BAR, capable of cleaving the C-glucosyl of barbaloin was isolated from human feces. This bacterium grew in PYF medium containing barbaloin and produced enzyme(s) that cleave(s) the C-glucosyl. The induction of the enzymes was completely inhibited in the presence of D-glucose. Nojirimycin inhibited the enzyme activity induced by barbaloin but it did not inhibit the bacterial growth in the presence of D-glucose. PMID- 8234420 TI - A note on the complex metabolism of danthron infused into the rat. AB - Danthron infused intravenously in rats shows a complex dose-dependent pattern of metabolism and excretion. The metabolites, particularly the more polar ones, are in general excreted predominantly in bile, to a lesser extent in urine. They can be separated as metabolite groups according to polarity and molecular size on a Sephadex LH 20 column. The present paper describes a further study within a bile derived metabolite group, which proved to be particularly heterogeneous. It contained more than a dozen metabolites, which were conjugates of four different aglycons including the parent danthron. 1H NMR spectral data for danthron monosulfate and monoglucuronide are also presented. PMID- 8234421 TI - Adverse effects of laxatives: fact and fiction. AB - Laxatives are generally well tolerated and may be considered safe drugs. When taken at much higher than the recommended doses (laxative abuse) some side effects may occur (e.g. hypokalemia, metabolic alkalosis, renal tubular damage). It is controversial whether the laxatives currently used may lead to morphologic changes of the autonomous nervous system of the colon. Melanosis coli is due to pigment-laden macrophages within the submucosa. It occurs after long-term intake of anthraquinones and has no functional consequences. No case of 'cathartic colon' has been observed during the last few decades, and we can assume that it was probably caused by laxatives which are no longer in use. PMID- 8234422 TI - Sennoside-induced secretion and its relevance for the laxative effect. AB - The effect of oral treatment with sennosides (50 mg/kg) on the time-course of net H2O and electrolyte transport rates was studied in 1-hour incubation experiments in the rat colon in vivo. Net H2O, Na+ and Cl- absorption rates did not change during the first 4 h after treatment, but were reversed to net secretion after 6 h and partly recovered during the next 18 h. K+ and Ca2+ were secreted in controls, and net secretion increased from 6 to 24 h after treatment. Paracellular permeability of [14C]erythritol was 3-fold 6 h after treatment but unchanged at other times after treatment (2, 4, 12 or 24 h). LDH leakage into the lumen was not enhanced by treatment. Neither mucosal Na+, K(+)-ATPase activity nor cAMP or phosphodiesterase activity was affected by sennosides. As stool consistency and acceleration of transit by sennosides has entirely normalized 24 h after treatment but not net absorption of H2O and electrolytes, it is concluded that there may be regional differences in the absorptive behavior of the colon induced by sennosides. Slow transit and increased absorption in some parts of the colon may overcome secretion in other parts. PMID- 8234423 TI - Effects of long-term sennoside treatment on in vitro motility of rat colon. AB - Colonic motility of rats chronically pretreated with either 10 or 40 mg sennosides/kg body weight daily, 30 mg sennosides/kg twice a week or 500 mg danthron/kg daily for 6 months has been evaluated in ex vivo preparations. Registration of colonic contractions 48 h after the last application of drugs or placebo revealed striking differences between the ascending, transverse and descending parts of the colon as well as between corresponding circularly or longitudinally orientated segments. Treatment did not influence motility in the various parts of the colon except some reduction in spontaneous motility in the descending colon. Response to electrical stimulation and acetylcholine was independent of treatment in most segments, but a decrease in maximal contraction was observed by the high daily sennoside dose and by danthron in the ascending colon and descending colon, respectively. Sensitivity to rhein, an active metabolite of sennosides, was similar in all groups. Thus, chronic treatment with anthranoids reveals no abnormal motility pattern but a residual pharmacological activity 48 h after the last administration of high doses. PMID- 8234424 TI - In vivo motility of rat colon chronically pretreated with sennosides. AB - Ceco-colonic myoelectrical activity was investigated in rats pretreated for 23 weeks by sennosides (10 or 40 mg/kg/day), Na-picosulfate (2.5 or 10 mg/kg/day) or laxative vehicle (control). On the last week of treatment the animals were equipped with Nichrome electrodes on the cecum, the proximal and distal colon. In comparison with controls, sennoside or Na-picosulfate treatment did not induce any significant (p > 0.05) change in the duration of long spike bursts (LSB) which are associated with phasic contractions. On the last 2 days of treatment the frequency of LSB for 2 h before and 2 h after laxative administration, as well as for 30 min after a 3-gram meal was not significantly (p > 0.05) different in control and treated animals. Similarly, on the first 2 days, as well as on days 13 and 14, after the end of treatment, no significant (p > 0.05) difference in the LSB frequency appeared between control and treated animals, in the fasted state or after a 3-gram meal. It is concluded that long-term treatment with sennosides or Na-picosulfate does not induce chronic changes in colonic motility in rats. PMID- 8234425 TI - Chronic sennoside treatment does not cause habituation and secondary hyperaldosteronism in rats. AB - Rats were treated with sennosides (6 x 10, 6 x 40 or 2 x 30 mg/kg weekly) or with danthron (6 x 500 mg/kg weekly) for 6 months. The laxative effect as measured by faecal wet weight during the first 10 h after treatment increased 3- to 4-fold by the higher sennoside doses (daily or intermittently) and 1- to 3-fold by danthron. The low sennoside dose had no measurable effect except on the 1st day (2 fold) compared with the control group. Mean faecal water content increased from 53% (controls) to 66-79% in rats treated with high sennoside doses and to 57 (1st day) -69% in danthron-treated rats. Serum aldosterone levels and mucosal Na(+)-K(+)-ATPase activities in the small intestine and colon did not change with treatment. There were no signs of habituation or secondary hyperaldosteronism due to sennosides or danthron in spite of chronic diarrhoea over 6 months. PMID- 8234426 TI - Light and electron-microscopic changes in the colon of the guinea pig after treatment with anthranoid and non-anthranoid laxatives. AB - Male guinea pigs were treated with senna pods, sennosides, danthron or bisacodyl by gastric intubation for 14 consecutive days. The animals were then killed and the intestine was evaluated by light and electron-microscopic techniques. Macroscopically, the mucosa of the cecum and upper colon was brown in color in the animals receiving anthranoid laxatives. No visible changes were detectable in the bisacodyl group. Evaluation by light and electron microscopy revealed cytoplasmic degeneration and increased apoptosis in the colonic surface epithelium after all laxatives. Most of the resulting apoptotic bodies were found in the lamina propria or had been phagocytosed by stromal macrophages, where they had been transformed into dark brown (anthranoids) or grey to light brown (bisacodyl) pigments. The intestinal changes were most pronounced in the cecum and decreased towards the distal part of the colon. It can be concluded that the morphological changes in the large intestine were similar in anthranoid and non anthranoid treatment, with the exception that the pigments found in macrophages differed in color and were therefore not always detectable macroscopically. PMID- 8234427 TI - The genotoxicity status of senna. AB - Genotoxicity tests were performed by several laboratories with the drug fructus sennae, senna extract, sennosides, rhein and aloe-emodin. The drug fructus sennae, the sennosides and rhein did not increase mutation frequencies in the following test systems: bacterial systems (Salmonella reverse mutation test and/or Escherichia coli forward mutation test); mammalian cell cultures [hypoxanthine guanine phosphoribosyl transferase (HGPRT) test; mouse lymphoma test; chromosome aberration test with Chinese hamster ovary cells]; bone marrow (micronucleus test; chromosome aberration test); melanoblast cells (mouse spot test) of rodents. With aloe-emodin mutagenic effects were observed only in vitro in the chromosome aberration test with CHO cells and in the Salmonella reverse mutation test (frameshift mutations in strains TA 1537, TA 1538 and TA 98). In the in vitro gene mutation test with V79 cells (HGPRT test) no mutagenic potential of aloe-emodin was observed. In in vivo studies [micronucleus test with bone marrow cells of NMRI mice, chromosome aberration test with bone marrow cells of Wistar rats, mouse spot test (crossing DBA/2J x NMRI) no indication for a mutagenic activity of aloe-emodin was found. The relevance of the absence of a mutagenic potential in in vivo test systems was strengthened by the fact that aloe-emodin could be found in the blood serum after oral administration. Additional information on the interaction of aloe-emodin with DNA was obtained from an ex vivo unscheduled DNA synthesis test performed with hepatocytes of male Wistar rats: aloe-emodin did not induce unscheduled DNA synthesis as expression of DNA damage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234428 TI - Effects of 'contact laxatives' on intestinal and colonic epithelial cell proliferation. AB - Experimental studies indicate that laxatives may induce epithelial damage. In addition, some laxatives induce the release of prostaglandins. Epithelial cell damage and release of prostaglandins are two pathways by which epithelial cell proliferation could be influenced. Furthermore, fermentable laxatives like lactulose may influence large intestine cell proliferation by the trophic effect of the fermentation products such as short-chain fatty acids. For these reasons an in vivo study in rats was performed to compare the short- and long-term effect of sennosides, bisacodyl, sodium picosulfate and lactulose on epithelial cell proliferation in the ileum and large intestine. Cell proliferation was examined by the BrdUrd labelling technique after 2, 6 and 12 weeks of continuous treatment. Studies in control animals show that the Labeling Index (LI) is higher in the cecum compared with other segments of the colon, and higher in the ileum than in the colon. Treatment with sennosides, bisacodyl and sodium picosulfate does not influence the LI in the ileum and induces no statistically significant increase of the LI when the treated groups are compared with the control group. The proliferative pattern along the crypts remains unchanged with all the laxatives throughout the study. It appears therefore that 'contact' laxatives have no major influence on ileal and colonic epithelial cell proliferation and should not be regarded as tumor-promoting substances. PMID- 8234429 TI - The senna drug and its chemistry. AB - Senna consists of the dried leaflets or fruits of Cassia senna (C. acutifolia) known in commerce as Alexandrian senna and of Cassia angustifolia commonly known as Tinnevelly senna. The senna plants are small shrubs of Leguminosae cultivated either in Somalia, the Arabian peninsula and near the Nile river. Tinnevelly senna is obtained from cultivated plants mainly in South India and Pakistan. Owing to the careful way in which the plant is harvested, the leaflets of the drug are usually little broken. Damaged leaves and lower quality products are often used for making galenicals. The senna pods (fruits) are collected during the same period as the leaves, then dried and separated into various qualities. The active principle of Senna was first isolated and characterized by Stoll in 1941. The first two glycosides were identified and attributed to the anthraquinone family. These were found to be dimeric products of aloe emodin and/or rhein which were named sennoside A and sennoside B. They both hydrolyze to give the aglycones sennidin A and B and two molecules of glucose. Later work confirmed these findings and further demonstrated the presence of sennosides C and D. Small quantities of monomeric glycosides and free anthraquinones seem to be present as well. The active constituents of the pods are similar to those of the leaves but present in larger quantities. Two naphthalene glycosides isolated from senna leaves and pods are 6-hydroxymusicin glucoside and tinnevellin glucoside. Both compounds can be utilized to distinguish between the Alexandrian senna and the India senna, since tinnevellin glucoside is only found in the latter and the first only in the C. senna.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234430 TI - Sennosides and aloin do not promote dimethylhydrazine-induced colorectal tumors in mice. AB - In a model of dimethylhydrazine-induced colorectal tumors in male mice aloin- or sennoside-enriched diets (0.03%) did not promote incidence and growth of adenomas and carcinomas after 20 weeks. Furthermore, in anthranoid-fed mice no significant changes in serum electrolytes as well as parameters of hepato- and nephrotoxicity were observed. PMID- 8234431 TI - Two-year carcinogenicity study with sennosides in the rat: emphasis on gastro intestinal alterations. AB - A carcinogenicity study was conducted by administering a purified senna extract via the drinking water to Sprague-Dawley rats of each sex for 2 years. The daily doses received were 0, 5, 15 and 25 mg/kg. Histopathological examination was restricted to tissues from the gastro-intestinal tract, liver, kidneys, adrenals and from tissues with any observed abnormalities or masses. A laxative effect was observed in high-dose females, and in mid- and high-dose males. No significant differences in survival were found between treated and control groups. Mean body weight gain was significantly decreased in high-dose males. Increased kidney weights were noted in mid-dose males and females, and high-dose females. Histopathological examination of control and high-dose rats did not indicate any difference in the incidence of neoplastic lesions. As regards non-neoplastic lesions, a treatment- but not dose-related increase in reactive mesenteric lymph node hyperplasia was observed in preterminally sacrificed rats. However, a corresponding increase was not noted in the terminally sacrificed rats or when preterminal and terminal animals were combined. No ultrastructural changes in the myenteric nerve plexus of the colon and jejunum could be detected in the small number of investigated tissue samples. In conclusion, results from the present investigation do not indicate any relationship between long-term administration of purified senna extract and gastrointestinal, liver, kidney or adrenal tumors in the rat. PMID- 8234432 TI - Clinical management of constipation. AB - First, it is important to find out whether the patient is complaining of infrequent defaecation, excessive straining at defaecation, abdominal pain or bloating, a general sense of malaise attributed to constipation, soiling, or a combination of more than one symptom. Second, one must decide if there is a definable abnormality as a cause of the symptom(s). Is the colon apparently normal or is its lumen widened (megacolon)? Is the upper gut normal or is there evidence of neuropathy or myopathy? Is the ano-rectum normal or is there evidence of a weak pelvic floor, mucosal prolapse, major rectocele, an internal intussusception or solitary rectal ulcer? Is there any systemic component such as hypothyroidism, hypercalcaemia, neurological or psychiatric disorder or relevant drug therapy? Choice of treatment will depend on this clinical evaluation. The range of treatments available is: Reassurance and stop current treatment: Patients with a bowel obsession may take laxatives or rectal preparations regularly without need. Increase dietary fibre: Most cases of 'simple' constipation respond to increased dietary fibre, possibly with an added supplement of natural bran. Toilet training and altered routine of life: Young people particularly may need to recognise the call to stool and alter their daily routine to permit and encourage regular defaecation. Medicinal bulking agent: Ispaghula, methyl cellulose, concentrated wheat germ or bran, and similar preparations are useful when patients with a normal colon find it difficult to take adequate dietary fibre. These preparations increase the bulk of stool and soften its consistency. They may be useful for those patients with the constipated form of irritable bowel syndrome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234433 TI - Suppression of the purgative action of rhein anthrone, the active metabolite of sennosides A and B, by calcium channel blockers, calmodulin antagonists and indomethacin. AB - The involvement of Ca2+ in the mechanism of the purgative action of rhein anthrone was studied. Among individual or combination pretreatments with calcium channel blockers, calmodulin antagonists and prostaglandin biosynthesis inhibitors, the combination of indomethacin and nifedipine completely blocked the diarrhoea induced by rhein anthrone and also inhibited its effects on colonic fluid and electrolyte transport, and large intestinal motility. Calmodulin antagonists were less active regarding suppression of the effects of rhein anthrone. We concluded that, in addition to prostaglandins, diarrhoea induced by rhein anthrone must also involve the calcium channel which can be blocked by nifedipine, but not verapamil. PMID- 8234434 TI - Constipation and cathartics as risk factors of colorectal cancer: a meta analysis. AB - Since individual case-control studies have failed to resolve the question whether constipation and use of cathartics represent significant risk factors of colorectal cancer, a meta-analysis was performed. The method by Peto was used to calculate pooled odds ratios of the cancer risk among exposed and unexposed subjects. The analysis of 14 previously published case-control studies revealed statistically significant risks for colorectal cancer associated with both constipation and use of cathartics, the pooled odds ratios and their 95 percent confidence intervals being 1.48 (1.32-1.66) and 1.46 (1.33-1.61), respectively. The increased risk applied similarly to both sexes, it was higher in cancer of the colon than rectum. Since constipation and cathartics are associated with much lower odds ratios than various dietary components, such as fat, meat, alcohol, and low-vegetable or low-residue diets, it appears that their risk reflects the confounding influence of underlying dietary habits. PMID- 8234435 TI - Retrospective study on laxative use and melanosis coli as risk factors for colorectal neoplasma. AB - In a retrospective study a cohort of 2,277 patients was defined by colonoscopy. Among other factors it was tested whether in these patients laxative use or the endoscopically diagnosed presence of melanosis coli were risk factors related to colorectal neoplasma. In comparison to patients taking no laxatives there was no significant increase in colorectal cancer rate either in laxative users or in patients with melanosis coli. However, there was a statistically significant association between the occurrence of colorectal adenomas and laxative use (relative risk of all patients exposed to laxatives = 1.72; of patients exposed to laxatives without melanosis coli = 1.47). The relative risk of adenoma development in patients with melanosis coli was 2.19. Taking into account that polyps can be diagnosed in the dark mucosa of melanosis coli patients more easily, even this relative risk of 2.19 seems to be related to a generally enhanced risk of laxative intake rather than to a special group of (anthranoid containing) laxatives. PMID- 8234436 TI - Influence of senna, fibre, and fibre + senna on colonic transit in loperamide induced constipation. AB - Retarded colonic transit and disturbed defecation are the most prominent pathophysiological mechanisms in constipation. Both may be influenced by bulking agents and by laxatives such as senna. Direct measurements of the influence of such substances on colonic transit are rare mainly because of technical problems. We measured gastric emptying, small and large intestinal transit in 24 healthy volunteers by a newly developed method employing a metal detector. Twelve persons taking a normal diet received loperamide in a dose sufficient to double the individual transit time. All subjects measured gastrointestinal transit time under normal conditions and with Sennatin containing purified sennosides 20 mg, Agiocur (30 g) as a fibre product containing 20 g Plantago ovata seeds/husks, or Agiolax (10 g) as a combination of 5.4 g P. ovata seeds/husks + 1.2 g senna pod with a sennoside content of 30 mg. Colonic transit was reduced by Sennatin and by Agiolax from 39 +/- 4 h to 17 +/- 3 h (p < 0.005). Agiocur did not influence colonic transit (39 +/- 3 h). Loperamide prolonged colonic transit from 27 +/- 0.7 to 72 +/- 12 h. This effect was abolished by Sennatin (30 +/- 5 h) and Agiolax (27 +/- 1 h) (p < 0.005), but not by Agiocur (64 +/- 13 h). The same effects were seen when right and left colonic transit were analyzed separately. Neither gastric emptying nor small intestinal transit were affected by either substance. All of the three study drugs increased stool weight significantly (p < 0.05). When stool frequency and consistency were compared, the effects were less clear.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234437 TI - A comparison of Agiolax and lactulose in elderly patients with chronic constipation. AB - In a double-blind crossover study the efficacies of Agiolax, a combination of fibre and senna pod, and lactulose were compared in 77 long-stay elderly patients with chronic constipation. Mean daily bowel frequency, stool consistency and ease of evacuation were significantly greater with Agiolax than lactulose. The recommended dose was exceeded more frequently with lactulose than Agiolax (chi 2 = 8.38, p < 0.01). Adverse effects were not different for the 2 treatments. In long-stay elderly patients with chronic constipation Agiolax and lactulose were well tolerated, but Agiolax proved a more effective treatment. PMID- 8234438 TI - Safety and efficacy of a bulk laxative containing senna versus lactulose in the treatment of chronic constipation in geriatric patients. AB - Thirty geriatric long-stay patients aged 65-94 years (mean 81.8) participated in the trial the aim of which was to examine bulk laxative plus senna (Agiolax) in the treatment of chronic constipation using lactulose (Levolac) as a reference medicine. Bulk laxative plus senna (daily doses 14.8 g) produced more frequent (p < 0.05) bowel habits (4.5 vs. 2.2-1.9/week) than lactulose (daily doses 20.1 g). Both laxatives proved to be safe to use. Our study indicated bulk laxative plus senna to be more efficient in treating constipation in geriatric long-stay patients. PMID- 8234439 TI - How effective is nonlaxative treatment of constipation? AB - In almost any textbook of medicine so-called simple measures are proposed for treating constipation, such as increasing physical activity, drinking more fluid, triggering the gastrocolonic response by, e.g., drinking a glass of water or a cup of coffee, and bowel training. All of these have not been formally evaluated in constipated patients. Biofeedback training is useful for paradoxical puborectal contraction ('anismus'). Dietary fibre, e.g. in the form of wheat bran, increases stool weight and decreases colonic transit time. So every patient should have an empiric treatment with an adequate amount of fibre. Some patients, especially those with slow transit constipation and those with stasis in the right hemicolon, are not likely to respond to a therapy with bran, however. PMID- 8234440 TI - In vitro effects of sennoside on contractile activity and fluid flow in the perfused large intestine of the rat. AB - The effect of sodium rhein on contractile activity and fluid flow in the rat complete large intestine was studied in vitro. Contractile activity was recorded using serosal strain gauges and volume transducers recorded distal fluid flow from the segment. Luminal sodium rhein (1 mM) produced a protracted increase in caecal activity yet increased colonic contractility transiently. Fluid flow from the preparation was increased and the number of propagated complexes was elevated after the initial 10 min of exposure. The effect did not appear to be related directly to dose. Sodium rhein (0.1 mM) did not significantly stimulate contractility and a higher dose (5 mM) only produced a transient effect on propagated contractions. However, this dose had the effect of significantly reducing activity when the rhein was replaced by normal buffer. The data suggest that the action of sodium rhein is subtle; after an initial excitation, the glycoside shifts the pattern of motor activity in favour of propulsion at the expense of segmentation. The large intestine is more able, therefore, to expel luminal contents in a caudal direction following the addition of this anthraquinone laxative. PMID- 8234441 TI - Anthranoids and the mucosal immune system of the colon. AB - The mechanism of action of anthranoids in general and of sennosides at the cellular level is not precisely known. Pseudomelanosis or pseudolipofuscinosis, a condition characterized by the accumulation of pigmented macrophages in the lamina propria, is one of the well-known effects of these products. It is most probably the result of an interaction between apoptotic epithelial cells and the lamina propria cellular infiltrate. Treatment of cell suspensions of intestinal epithelial cells and of human intestinal epithelial cells in culture with rhein anthrone, the active compound of sennosides, demonstrates a direct influence of the drug on these epithelial cells. Low doses induce alterations in cellular shape and organelles consistent with increased metabolism. High doses induce apoptotic changes. The interaction between the epithelial cells and cells of the monocyte/macrophage lineages induces also the release of prostaglandins of the E series as shown by experiments on cell cultures of epithelial cells and peripheral blood cells. An increase of PGE2 release to about 140% of the control value is noted following administration of low doses of rhein anthrone to a combination of human intestinal epithelial cells and human peripheral blood mononuclear cells. This finding indicates that rhein anthrone is activating cellular components of the intestinal immune system and may by this pathway induce secretion and motility. PMID- 8234442 TI - Effect of senna is not mediated by platelet-activating factor. AB - The effect of in vivo treatment with senna was examined on the ex vivo formation of platelet-activating factor (PAF) by small and large intestine of rat, mouse and guinea pig. A single or a prolonged oral administration of senna (60-240 mg/kg) to animals did not increase intestinal PAF content. Nor did senna increase the intraluminal release of acid phosphatase. A similar result was obtained in the colonic tissue of rat perfused in vitro with rhein (1-500 micrograms/ml) or rhein anthrone (1-500 micrograms/ml). In contrast, a single oral administration of phenolphthalein (20 mg/kg), bile salts (20 mg/kg) or magnesium sulfate (30 mg/kg) to rats increased intestinal PAF content. Magnesium sulfate also increased the intraluminal release of acid phosphatase. Colonic tissue of rats perfused in vitro with calcium ionophore A23187 (10 micrograms/ml) formed large amounts of PAF and acid phosphatase. PAF stimulates intestinal motility and secretion and mediates gut damage while acid phosphatase is a marker of cellular damage. Therefore, our data suggest that senna is well tolerated in animals and PAF does not mediate senna-induced laxation. PMID- 8234443 TI - Serotonin antagonists inhibit sennoside-induced fluid secretion and diarrhea. AB - The aim of this study was to investigate whether 5-hydroxytryptamine (serotonin, 5-HT) is involved in the mediation of sennoside-induced colonic fluid secretion and diarrhea. Oral administration of purified sennosides (25, 40 and 64 mg/kg) dose-dependently reversed net fluid absorption to net fluid secretion, enhanced the incidence of diarrhea and stimulated the release of 5-HT into the colonic lumen from 7.1 to 17.3 ng/g wet weight. The 5-HT2 antagonist ketanserin and the 5 HT3 antagonist tropisetron dose-dependently but only partially reduced sennoside (40 mg/kg)-induced fluid secretion whereas the 5-HT3 antagonist granisetron dose dependently reduced and at 300 micrograms/kg totally abolished sennoside-induced secretion. Granisetron, but not ketanserin and tropisetron, reduced the incidence of diarrhea in sennoside-treated rats, indicating the involvement of 5-HT also in acceleration of large intestinal transit. It is concluded that 5-HT is an important mediator both of sennoside-induced fluid secretion in the rat colon and of diarrhea. PMID- 8234444 TI - Neuro-immunophysiology of colon function. AB - Neuroimmunophysiology involves direct communication between the mucosal immune system and the minibrain in the colon. Communication is chemical in nature (paracrine) and involves specialized sensing functions of the immune cells for specific antigens together with intelligent interpretation of the signals by the enteric nervous system. Immunoneural integration progresses sequentially starting with immune detection followed by signal transfer to enteric microcircuits followed by neural interpretation and then selection of a specific neural program of coordinated mucosal secretion and motor propulsion that acts to clear the antigenic threat from the colonic lumen. Histamine and serotonin are important chemical messages released from mucosal mast cells to act at specific receptor subtypes on neural elements of the enteric minibrain. PMID- 8234445 TI - Rhein stimulates electrogenic chloride secretion by activation of submucosal neurons in guinea pig colon. AB - Conventional flux chamber methods were applied to investigate the mode of action of rhein, an active metabolite derived from colonic microbial fermentation of the naturally occurring sennoside laxatives, in muscle-stripped segments of guinea pig colon. Mucosal or serosal application of rhein (10 nmol/1 to 0.5 mmol/l) resulted in a dose-dependent increase in short-circuit current (Isc) that was superimposed by irregular fluctuations in Isc. The response to electrical field stimulation was increased. The rhein-evoked increase in Isc was reduced by serosal addition of 50 mumol/l bumetanide, 1 mumol/l tetrodotoxin, 1 mumol/l atropine and 10 mumol/l piroxicam but not 100 mumol/l hexamethonium, 1 mumol/l ICS 205 930 or 10 mumol/l cimetidine. The study suggests that rhein activates chloride secretion by excitation of submucosal neurons and release of acetylcholine and endogenous prostaglandins, but not by release of histamine or serotonin. PMID- 8234446 TI - Antioxidant and radical scavenging effects of anthraquinones and anthrones. AB - The activity of seven anthraquinones and four anthrones against nonenzymatic and enzymatic lipid peroxidation in vitro and their ability to scavenge free radicals have been studied. In nonenzymatic peroxidation in rat hepatocytes induced by t butyl hydroperoxide, dithranol and anthrone were the strongest antioxidants, having IC50 values of 8 +/- 1 and 24 +/- 5 mumol/l, respectively. Rhein (IC50 64 +/- 2 mumol/l) and aloe-emodin (IC50 65 +/- 3 mumol/l) showed the highest inhibitory activity against peroxidation of linoleic acid catalyzed by soybean 15 lipoxygenase. Anthrone (IC50 62 +/- 2 mumol/l), dithranol (IC50 72 +/- 1 mumol/l) and rhein anthrone (IC50 76 +/- 6 mumol/l) were the most effective radical scavengers of the diphenylpicrylhydrazyl radical. The antioxidant activities in hepatocytes and the radical scavenging activities were correlated, whereas the inhibition of enzymatic lipid peroxidation showed no correlation with the two other effects. PMID- 8234447 TI - Metabolism and pharmacokinetics of anthranoids. AB - Anthranoid derivatives are used all over the world as a treatment for constipation. These compounds are present in several drugs of plant origin, especially as O- or C-glycosides. Besides featuring different substituents, the aglycone might consist of an anthraquinone, an anthrone or a dianthrone. So far, detailed information concerning their metabolism and pharmacokinetic characteristics is available only in a few cases. The best characterized compounds are sennoside, a dianthrone O-glycoside present in senna leaves and senna pods, and its aglycone (rhein anthrone). After oral administration, sennoside is degraded only in the lower parts of the gastrointestinal tract, releasing its active metabolite rhein anthrone. Nowadays, this process is understood at the molecular level. A study with 14C-labelled rhein anthrone administered intracecally to rats, revealed that the compound is scarcely absorbed. Since on the contrary its anthraquinone equivalent is absorbed to a much larger extent, it is inferred that dianthrone- or anthrone-glycosides exhibit a lower systemic availability than anthraquinone O-glycosides. PMID- 8234448 TI - Analytics of senna drugs with regard to the toxicological discussion of anthranoids. AB - Toxicological studies indicate that two hydroxyanthraquinones (HAs), aloe-emodin and emodin, present as minor components in senna, might represent a genotoxic or cancerogenetic risk for man. Since aloe-emodin and emodin occur in senna in the free form as well as their glucosides and dianthrone glucosides, a HPLC method was established to allow the quantification of all free and glycosidic 1,8 dihydroxy anthranoids. The sum of the free HAs and their calculated content in each of their prodrug forms is defined as the potential HA content. For the comparison of different senna drugs in respect to the genotoxic risk arising from their potential aloe-emodin or emodin contents, a risk index has been established. PMID- 8234449 TI - Benzodiazepine substitution in medical practice. Analysis of pharmacoepidemiologic data based on expert interviews. AB - Pharmacoepidemiologic data have shown a consistent reduction in benzodiazepine (BZ) prescriptions over the past decade. The question remains as to whether BZs are simply put aside or whether other medications are used as substitutes. Expert interviews, at which a stratified sample of 114 psychiatrists, internists, and general practitioners were presented with case studies, were conducted to learn about the therapeutic alternatives in the field of benzodiazepine-related indications administered in daily practice. These results were used to analyze trends in pharmacoepidemiologic prescription data for all patients under the general health insurance plan from 1981 to 1988. The experts identified distinct alternatives to BZs in different clinical situations, including neuroleptics, antidepressants, phototherapeutics, and analgesics. When these findings were transferred to pharmacoepidemiologic data, results revealed an increase in the prescription of alternative medications that apparently compensated for reduced BZ use. Overall there was no change (or, rather, no increase) in the total of psychotropic prescriptions during the period of reduced BZ prescriptions. Our findings indicate that reduction in benzodiazepines prescription is associated with substitution by various other psychotropic drugs. This has positive as well as negative consequences, and there must be discussed in detail before sound recommendations can be given as to which type of drug. BZs included, is the best choice in which type of illness. PMID- 8234450 TI - Benefits and risks of psychotropic medication in the eyes of the general public: results of a survey in the Federal Republic of Germany. AB - In 1990, a survey of the population of the FRG, including the new states, was carried out to investigate the attitude of the general public toward psychotropic drugs. This showed that they are regarded with fare more suspicion than trust. Their use in the treatment of mental disorder was rejected twice as often as it was recommended. Those questioned submitted three main arguments against the use of psychotropics: (1) they can cause unwanted effects, in particular dependency; (2) their action is restricted, i.e., they only treat the symptoms and not the cause of the illness; (3) they are ineffective, having either no effect or effects of doubtful or temporary nature. The use of psychotropic agents was recommended, if at all, for the purpose of sedating the patients, providing relief from their problems, and lessening their symptoms. PMID- 8234451 TI - Tricyclic-induced orthostatic hypotension. Significant difference in depressed and non-depressed states. PMID- 8234452 TI - Late onset tricyclic-induced agranulocytosis with an atypical course. PMID- 8234453 TI - Lorazepam in treatment of catatonic schizophrenia. PMID- 8234454 TI - Inappropriate patient sexual behaviors. PMID- 8234455 TI - The use of a mechanical model to describe the stiffness and damping characteristics of the knee joint in healthy adults. AB - BACKGROUND AND PURPOSE: Stiffness is a common clinical complaint but is rarely quantified by clinicians. The purpose of this study was to determine the reliability of a relaxed oscillation test that yields stiffness and damping coefficients of the knee. These coefficients describe the knee joint's resistance to bending and the time-dependent nature of that resistance. SUBJECTS AND METHODS: Effects of age, gender, and knee position on these coefficients were assessed in 96 healthy volunteers aged 20 to 79 years. Measures were based on the premise that the knee joint can be modeled as a damped spring. Oscillations of the knee were recorded using an electrogoniometer with the knee oscillating through about 45 and 75 degrees of knee flexion. RESULTS: Intraclass correlation coefficients revealed moderate to high reliability in the measurements taken three times in the same test session and on three separate days. Analysis of variance showed significant increases in stiffness and damping coefficients in the male subjects as compared with the female subjects. Analysis of variance also suggested an age effect on stiffness coefficients at the 75-degree test position, with decreasing stiffness with age. Both stiffness and damping coefficients were significantly smaller when measured at the 75-degree test position compared with the 45-degree position (Student's t test). CONCLUSION AND DISCUSSION: These results demonstrate a reliable method of measuring knee joint stiffness, and they correlate well with known morphological differences related to age and gender. This measure may prove to be more useful in evaluating the function of the knee than more commonly used assessments. It may also lead to a better understanding of how the knee functions in such activities as locomotion. PMID- 8234456 TI - Factors associated with burnout of physical therapists in Massachusetts rehabilitation hospitals. AB - BACKGROUND AND PURPOSE: The purpose of this study was to determine the factors associated with burnout in physical therapists at rehabilitation hospitals. Burnout is a serious concern because it can lead to psychosomatic complaints, work-associated withdrawal behaviors, and a decreased quality of care. SUBJECTS: Full-time, nonadministrative, nonsupervisory, staff physical therapists working in inpatient settings in rehabilitation hospitals in Massachusetts were surveyed. METHODS: The survey included demographic data, the Maslach Burnout Inventory (MBI), and questions designed to address attributes of the work environment and individual personalities. The MBI scores were calculated according to instructions provided by publishers to determine the degree of burnout experienced by this group. Questions involving personality and work environment were submitted to factor analyses. Multivariate analyses were done to determine the factors related to burnout. RESULTS: Forty-six percent of the respondents scored high on the emotional exhaustion subscale of the MBI, 20% scored high on the depersonalization subscale, and 60% scored low on the personal accomplishment subscale. As a whole, the sample demonstrated moderate burnout. Three factors emerged from the factor analysis. The factors communication/connectedness, achievement, and time constraints accounted for 69% of the variability in emotional exhaustion and 73% of the variability in depersonalization and personal accomplishment. Burnout was not significantly associated with the therapists' number of years of practice, number of years on the job, or number of patients seen daily. CONCLUSION AND DISCUSSION: Recognition of factors contributing to burnout may prove to be instrumental in the prevention of burnout in physical therapists and the problems that arise from it. PMID- 8234457 TI - Experiences of student and practicing physical therapists with inappropriate patient sexual behavior. AB - BACKGROUND AND PURPOSE: The objectives of this study were (1) to describe the extent to which practicing physical therapists and physical therapy students have reported experiencing inappropriate patient sexual behavior (IPSB), (2) to document the consequences of IPSB, and (3) to identify the strategies the subjects have used to manage IPSB. SUBJECTS AND METHODS: A survey questionnaire was sent to 118 physical therapists and 87 physical therapy students. Completed questionnaires were returned by 74.1% of the subjects (over 70% in each group). RESULTS: The majority of respondents (80.9%) reported having encountered some level of IPSB. Although almost half of the physical therapists and one third of the third- and fourth-year students reported experiencing severe IPSB, including forceful sexual touching and deliberate sexual exposure, only 20% of the respondents perceived that they had been "sexually barassed." Most of the respondents were satisfied with the way in which they handled the IPSB; however, almost 90% considered that in-service and undergraduate education would be desirable. CONCLUSION AND DISCUSSION: It is concluded that this problem should be addressed by further study with a broader sample and by inclusion of these issues in education programs for students and physical therapists. PMID- 8234458 TI - Correlates of exercise compliance in physical therapy. AB - BACKGROUND AND PURPOSE: This correlational study describes factors that are related to patient compliance with exercise regimens during physical therapy. We investigated whether patient compliance was related to characteristics of the patient or the patient's illness, to the patient's attitude, or to the physical therapist's behavior. SUBJECTS AND METHODS: Of a random sample of 300 physical therapists in private practice in the Netherlands, 222 therapists responded to a questionnaire survey. Eighty-four respondents also made audio recordings. Materials of the study were 1,931 registration forms, 1,837 audio-recorded sessions of physical therapy sessions, and 1,681 patient questionnaires. RESULTS: The results show that the three main factors related to noncompliance were (1) the barriers patients perceive and encounter, (2) the lack of positive feedback, and (3) the degree of helplessness. The first factor, the barriers patients perceive and encounter, shows the strongest relation with noncompliance. The results also show that noncompliance is more strongly related to the characteristics of the illness than to the illness, a bad prognosis is negatively related to compliance, and much hindrance of the complaint is positively related to compliance. There was no difference between men and women with regard to patient compliance, but less educated patients were slightly more compliant than more highly educated patients. CONCLUSION AND DISCUSSION: These correlational findings can be used to formulate hypotheses of cause and effect in future clinical research. Future research should take into account the type and efficacy of therapeutic exercise for different diseases. For physical therapy practice, it seems important that physical therapists carefully explore which problems patients encounter in their efforts to comply and that they seek solutions to those problems in mutual cooperation with their patients. PMID- 8234459 TI - Perceptions of physical therapists toward patient education. AB - BACKGROUND AND PURPOSE: The purpose of this study was to assess the perceptions of physical therapists regarding their involvement in patient education. We developed a questionnaire to assess procedures or activities taught, methods or tools used, important physical therapist behaviors, techniques used to assess the effectiveness of teaching, barriers to delivering effective education, relative value of patient care activities as compared with other activities, and factors contributing to the development of teaching skills. SUBJECTS AND METHODS: The questionnaire was sent to 300 American Physical Therapy Association members nationwide chosen by a random sample. Two hundred questionnaires, or 69% of the eligible responses were used in the data analysis. RESULTS: Most respondents indicated they teach 80% to 100% of their patients. Nearly all respondents teach range-of-motion techniques, home programs, and treatment rationale through the use of demonstration and verbal discussion. Most respondents recognize the importance of many therapist behaviors related to patient teaching and use observation and evaluation to assess the effectiveness of their teaching. Barriers to patient education most frequently cited were psychological factors of the patient. Interaction with patients was considered most important to the development of respondents' teaching skills. CONCLUSION AND DISCUSSION: Implications and suggestions for further research are discussed. PMID- 8234460 TI - Twenty-seventh Mary McMillan Lecture. On passing from ignorance to knowledge. PMID- 8234461 TI - PTA supervision. PMID- 8234462 TI - Upper excited state photochemistry of DNA. AB - The quantum yields for cyclobutylpyrimidine dimers, alkali-labile sites, and frank strand breaks in double-stranded DNA have been measured using low-intensity radiation at 199.8, 217.8, and 239.5 nm from a Raman-shifted frequency quadrupled Nd:YAG laser. The quantum yield for cyclobutylpyrimidine dimers was also measured using 254 nm radiation from a low-pressure mercury lamp. The quantum yield for cyclobutylpyrimidine dimers is constant within a factor of two between 254 and 199.8 nm except for 239.5 nm, indicating that upper excited singlet states of bases convert efficiently to the lowest singlet state. The quantum yields for alkali-labile sites and frank strand breaks both increase as the wavelength decreases but follow different patterns. These results indicate that alkali labile sites from a higher excited state of the base, whereas frank strand breaks form by excitation of the sugar-phosphate backbone. PMID- 8234463 TI - The photochemistry of adenosine: intermediates contributing to its photodegradation mechanism in aqueous solution at 298 K and characterization of the major product. AB - The steady-state (254 nm) photolysis of 9-(beta-D-erythropentofuranosyl)adenine (adenosine) in aqueous solution was studied. Photodestruction yields on the order of 1.3 x 10(-3) were determined at room temperature by measuring the initial decrease in the absorption maximum as a function of irradiation time. The use of high performance liquid chromatography (HPLC) permitted a more exact determination of the yield (2.5 x 10(-3). The formation of photoproducts was also studied using HPLC. In the photolysis of 50 microM aqueous solutions of adenosine under anaerobic conditions at least 11 stable photoproducts are formed that absorb at 260 nm, the wavelength of maximum absorption of adenosine. The major photoproduct was also isolated and characterized as adenine; its formation yield was determined to be 4.5 x 10(-4). This yield is affected by the presence of oxygen and by the initial concentration of adenosine employed. Fluorescence emission and excitation spectra were used to monitor the formation of highly fluorescent photoproducts that emit with maxima at 365, 398, and 430 nm and absorb in the wavelength region of 240-380 nm. The reactive species in the photodestruction mechanism were established using substrates that react selectively with the respective short-lived species. Photoionization is a primary photoprocess implied by these studies. The triplet state of adenosine also contributes to the photodestruction mechanism. PMID- 8234464 TI - Benzophenone-sensitized lipid peroxidation in linoleate micelles. AB - Benzophenone-photosensitized peroxidation in linoleic acid micelles (Type I reaction) has been investigated and quantum yields of both sensitizer photolysis and lipid peroxidation have been obtained. Benzophenone as well as its derivative, benzophenone-4'-heptyl-4-pentanoic acid (BHPA) have been utilized as sensitizers in this study. A quantum yield of 1.2 for hydroperoxide formation, compared with a benzophenone disappearance quantum yield of 0.41 (in the same linoleate micellar solution) was obtained; yields of 1.3 and 0.6 were found for comparable systems in which BHPA was the sensitizer. Although such data are consistent with the peroxidation mechanism of chain reaction, it is necessary to know the yields of initiating species to arrive at chain reaction lengths in these systems. Such yields have been determined by flash photolysis. The dependencies of sensitizer photolysis and hydroperoxide yields on linoleate concentration have also been determined in order to elucidate the effects of micelle formation or molecular organization on the peroxidative process. Additionally, for comparison in a saturated surfactant (lipid) system hydrogen abstraction was investigated by photolysis techniques in sodium dodecyl sulfate alone using these two sensitizers. PMID- 8234465 TI - Singlet-oxygen generation at gas-liquid interfaces: a significant artifact in the measurement of singlet-oxygen yields from ozone-biomolecule reactions. AB - Several ozone-biomolecule reactions have previously been shown to generate singlet oxygen in high yields. For some of these ozone-biomolecule reactions, we now show that the apparent singlet-oxygen yields determined from measurements of 1270 nm chemiluminescence were artifactually elevated by production of gas-phase singlet oxygen. The gas-phase singlet oxygen results from the reaction of gas phase ozone with biomolecules near the surface of the solution. Through the use of a flow system that excludes air from the reaction chamber, accurate singlet oxygen yields can be obtained. The revised singlet-oxygen yields (mol 1O2 per mol O3) for the reactions of ozone with cysteine, reduced glutathione, NADH, NADPH, human albumin, methionine, uric acid and oxidized glutathione are 0.23 +/- 0.02, 0.26 +/- 0.2, 0.48 +/- 0.04, 0.41 +/- 0.01, 0.53 +/- 0.06, 1.11 +/- 0.04, 0.73 +/ 0.05 and 0.75 +/- 0.01, respectively. These revised singlet-oxygen yields are still substantial. PMID- 8234466 TI - Calcium-dependent fluorescence lifetimes of Indo-1 for one- and two-photon excitation of fluorescence. AB - We characterized the fluorescence intensity decays of Indo-1, which is commonly used as an emission wavelength-ratiometric calcium probe. The apparent lifetime of the long-wavelength side of the emission of Indo-1 is dependent on Ca2+. This long-wavelength emission displays the characteristics of an excited-state reaction, that is, a negative preexponential component in the multiexponential analysis. The emission spectra and lifetime of Indo-1 appear to be identical for one-photon and two-photon excitation at 351 and 702 mn, respectively, suggesting that the relative one- and two-photon cross sections are similar for the calcium free and calcium-bound forms of Indo-1. Also, the two-photon cross section of Indo-1 is relatively high, about 4 x 10(-49) cm4 s/photon molecule at 690 nm for both the calcium-free and calcium-bound forms. Hence, Indo-1 can be used for calcium imaging based on one- or two-photon excitation, using either emission wavelength ratios or lifetime imaging methods. PMID- 8234467 TI - Photobleaching of mono-L-aspartyl chlorin e6 (NPe6): a candidate sensitizer for the photodynamic therapy of tumors. AB - Most sensitizers used for the photodynamic therapy (PDT) of tumors photobleach on illumination. Thus, it is of interest to examine the photobleaching behavior of new sensitizers proposed for use in PDT. This report surveys the quantum yields and kinetics of the photobleaching of mono-L-aspartyl chlorin e6(NPe6), a hydrophilic chlorin that has many of the photoproperties desirable in a sensitizer for clinical PDT. It is a very effective sensitizer for the PDT of several types of model tumors in animals and is now in Phase I clinical trials. The quantum yield of NPe6 photobleaching in pH 7.4 phosphate buffer in air was 8.2 x 10(-4); this is greater than the yields for typical porphyrin photosensitizers. For example, the yields for hematoporphyrin and uroporphyrin are 4.7 x 10(-5) and 2.8 x 10(-5), respectively. The yield decreased significantly in organic solvents of low dielectric constant. The Sn derivative of NPe6 was more light stable than NPe6 (yield = 5.7 x 10(-6), while the Zn derivative was more sensitive (yield = 1.9 x 10(-2). Oxygen appeared to be necessary for the photobleaching of NPe6; however, bleaching was not inhibited by 100 mM azide, an efficient quencher of singlet oxygen. The photooxidizable substrates cysteine, dithiothreitol and furfuryl alcohol increased the quantum yield of photobleaching two- to four-fold, while the electron acceptor, metronidazole, increased it almost six-fold. Photobleaching yields for several other chlorins were also measured. PMID- 8234468 TI - Phthalocyanine-induced photohemolysis: structure-activity relationship and the effect of fluoride. AB - Phthalocyanine (Pc) containing A1, Ga or Zn as central metal ligand and substituted with a varying number of sulfonic acid residues as well as additional benzene rings were synthesized and their photodynamic activity was assayed using photohemolysis of human erythrocytes as an endpoint. The Pc derivatives varied > 300-fold in their photodynamic activity. Activity correlated with binding of the dye to the cell, with the exception of some of the amphiphilic dyes where cell uptake was an order of magnitude higher than expected from the observed activity. Fluoride was shown to inhibit A1PcSn-induced photohemolysis. This effect occurred also with other A1Pc and GaPc derivatives, but the concentration of F- required to slow photohemolysis by a factor of two (Ki) varied between 4 microM and 10 mM. Fluorescence spectral studies indicated complex formation between F- and the dye, which was stronger for A1Pc than GaPc derivatives. Ultrastructural studies using scanning electron microscopy showed that the photosensitized cells were converted to spherocytes and that F- prevented this to a large extent. PMID- 8234469 TI - Efficient photodynamic action of Victoria blue BO against the human leukemic cell lines K-562 and TF-1. AB - Photodynamic induced cytotoxicity by Victoria blue BO (VB-BO), merocyanine 540 (MC540), Nile blue A (NB) and 4-tetrasulfonatophenyl-porphyrin (4-TSPP) has been studied on two human leukemic cell lines: K-562 and TF-1. Cells were incubated with dyes and irradiated with different doses of white light. Cell survival was assessed by propidium iodide (PI) staining using flow cytometry analysis. Concentrations of 5 x 10(-8) M VB-BO were found to kill 75% of cells, and a concentration of 1 x 10(-7) M induced more than 99% of cell killing. To obtain the same cytotoxic level, the presence of 2.6 x 10(-5) M of MC540 during irradiation was needed. Under the conditions used, NB was ineffective as a photosensitizer, although uptake studies showed that this dye was taken by the cells in much greater amounts than any other studied dye. Cell cycle distribution of TF-1 cells, surviving MC540 or VB-BO photosensitization has been studied by flow cytometry analysis after staining with Hoechst 33342 and PI. It was found that cells in G1 phase were slightly more resistant toward MC540- and VB-BO mediated photosensitization than cells in other phases of the cell cycle. PMID- 8234470 TI - Dye-sensitized destabilization of liposomes bearing photooxidizable lipid head groups. AB - Liposomes were prepared from mixtures of dipalmitoyl-L-alpha-phosphatidylcholine and up to 40% mol:mol of N-stearoyl-L-histidine (NSH) in the presence of the hydrophobic sensitizer DHE. In the dark such liposomes are stable and retain entrapped salts. On photolysis with visible light, liposomes leak trapped ions at NSH concentrations greater than 10% mol:mol. Up to 15% mol:mol NSH concentration leakage is seen only during the illumination period, whereas at higher concentration the liposomes continue to leak contents after illumination and fuse to form larger structures. Photolysis of the liposomes is accompanied by oxygen uptake in proportion to the NSH concentration within the bilayer. Photocontrol of liposome permeability through oxidation of membrane additives such as NSH offers a potential means for controlled drug delivery and might be useful as an adjunct to photodynamic therapy. PMID- 8234471 TI - Spectroscopic studies of cutaneous photosensitizing agents. XVIII. Indomethacin. AB - The photochemistry, photophysics, and photosensitization (Type I and II) of indomethacin (IN) (N-[p-chlorobenzoyl]-5-methoxy-2-methylindole-3-acetic acid) has been studied in a variety of solvents using NMR, high performance liquid chromatography-mass spectroscopy, transient spectroscopy, electron paramagnetic resonance in conjunction with the spin trapping technique, and the direct detection of singlet molecular oxygen (1O2) luminescence. Photodecomposition of IN (lambda ex > 330 nm) in degassed or air-saturated benzene proceeds rapidly to yield a major (2; N-[p-chlorobenzyl]-5-methoxy-2-methyl-3-methylene-indoline) and a minor (3; N-[p-chlorobenzoyl]-5-methoxy-2,3-dimethyl-indole) decarboxylated product and a minor indoline (5; 1-en-5-methoxy-2-methyl-3- methylene-indoline), which is formed by loss of the p-chlorobenzoyl moiety. In air-saturated solvents two minor oxidized products 4 (N-[p-chlorobenzoyl]-5-methoxy-2-methyl-indole-3 aldehyde) and 6 (5-methoxy-2-methyl-indole-3-aldehyde) are also formed. When photolysis was carried out in 18O2-saturated benzene, the oxidized products 4 and 6 contained 18O, indicating that oxidation was mediated by dissolved oxygen in the solvent. In more polar solvents such as acetonitrile or ethanol, photodecomposition is extremely slow and inefficient. Phosphorescence of IN at 77 K shows strong solvent dependence and its emission is greatly reduced as polarity of solvent is increased. Flash excitation of In in degassed ethanol or acetonitrile products no transients. A weak transient is observed at 375 nm in degassed benzene, which is not quenched by oxygen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234472 TI - Stress protein expression in murine tumor cells following photodynamic therapy with benzoporphyrin derivative. AB - Photodynamic therapy (PDT) has been proven as a method of tumor eradication and is currently being used clinically to treat a wide variety of malignancies. Although it is understood that the interaction of light and sensitizer results in the production of potentially damaging oxygen species, the mechanism by which tumors are destroyed has yet to be defined fully. Using a new porphyrin sensitizer, benzoporphyrin derivative (BPD), we examined protein expression in murine tumor cells following treatment as an indication of molecular changes to target tissue concurrent with PDT-mediated damage. In order to assess the relevance of the results obtained using an in vitro PDT model, metabolic labeling of proteins synthesized subsequent to PDT was performed both in tumor cells grown and treated in tissue culture dishes and in cells explanted from PDT-treated solid tumors. We observed that the oxidative stress associated with PDT-resulted in the induction of a number of proteins corresponding to a set of heat-shock or stress proteins, and that the pattern of expression was similar when tumor cells were treated in vitro and in vivo. These results support the use of in vitro models in the dissection of the molecular effects of PDT and provide the foundation for future experiments that will examine the role of the immune system in tumor eradication by PDT. PMID- 8234473 TI - Biologically effective dose of solar ultraviolet radiation estimated by spore dosimetry in Tokyo since 1980. AB - The biologically effective dose of solar UV radiation has been measured in Tokyo since 1980 using Bacillus subtilis spores. To determine the cumulative dose in a half day, several samples of UV-sensitive spores were exposed in successive intervals from the solar-noon time. Because fluence-survival curves were exponential, the number of lethal hits received by the spores was calculated for each interval and termed inactivation dose (ID). The total number of hits obtained in a half day (half-day ID) was correlated with the amount of global insolation by a power-function regression. The regression analyses were performed for the data collected on 35 days from 1980 to 1986 and for the data collected on 53 days from 1989 to 1991. The latter data set yielded significantly larger estimates of half-day ID relative to the insolation than the former. These analyses suggested that the biologically effective dose relative to the insolation increased about 30% at some time in the later part of 1980s at this location. Changes of solar activity, air pollution and stratospheric ozone layer were considered as potentially responsible for this increase, but identification of the causative factors requires further efforts. PMID- 8234474 TI - The effects of thromboxane inhibitors on the microvascular and tumor response to photodynamic therapy. AB - Vascular stasis and tissue ischemia are known to cause tumor cell death in several experimental models after photodynamic therapy (PDT); however, the mechanisms leading to this damage remain unclear. Because previous studies indicated that thromboxane release is implicated in vessel damage, we further examined the role of thromboxane in PDT. Rats bearing chondrosarcoma were injected with 25 mg/kg Photofrin (intravenously) 24 h before treatment. Light (135 J/cm2, 630 nm) was delivered to the tumor area after injection of one of the following inhibitors: (1) R68070: a thromboxane synthetase inhibitor; (2) SQ 29548: a thromboxane receptor antagonist; and (3) Flunarizine: an inhibitor of platelet shape change. Systemic thromboxane levels were determined. Vessel constriction and leakage were evaluated by intravital microscopy. Tumor response was assessed after treatment. Thromboxane levels were decreased more than 50% with SQ-29548 as compared to controls. Thromboxane levels in animals given R68070 and Flunarizine remained at baseline levels. SQ-29548 and R68070 reduced vessel constriction compared to controls, while Flunarizine totally prevented vessel constriction. R68070 and SQ-29548 inhibited vessel permeability compared to PDT controls; Flunarizine did not. Animals given these inhibitors showed markedly reduced tumor cure. These results indicate that the release of thromboxane is linked to the vascular response in PDT. PMID- 8234475 TI - Relative ground and excited state energies of CH3(CH = CH)5CH = NC4H9, its hydrogen-bonded and proton-transferred species, and charge partitioning and distribution in the protonated Schiff base of retinal. AB - CH3(CH = CH)5CH = NC4H9 (compound 1) is structurally related to the Schiff base of retinal, the prosthetic group in visual pigments. Dilute solutions of a weak acid (phenol) and 1 in a hydrocarbon solvent, when subjected to decreasing temperature, show striking changes in electronic absorption spectra. Initially only the spectrum of compound 1 is present, but as the temperature is lowered, the absorbance of 1 decreases, and the spectrum of the H-bonded form of 1 appears and increases. Continued temperature lowering then causes a decrease in absorption of the H-bonded form and an appearance and rise in absorption of the proton-transferred form of 1. Concentrations of the various species are measured as a function of temperature, and by standard procedures, the thermodynamic constants for both reaction steps are computed. Values of delta H0 are taken as relative energies among the three ground states, and the lambda max value of each species yields relative energies among excited states. By employing data from electronic absorption spectroscopy, nuclear magnetic resonance (NMR) and theoretical calculations for retinal Schiff base, charge partitioning between nitrogen and the polyene chain and charge distribution among the carbon atoms of the polyene chain are calculated. PMID- 8234476 TI - The conformational analysis and photoisomerization of retinochrome analogs with polyenals. AB - 3,7-Dimethyl-2,4,6,8,10-dodecapentaenal was synthesized for reconstitution of the retinochrome analog. Its opsin shift was 1000 cm-1 smaller than that of native retinochrome, whose chromophore contains the same number of double bonds. The conformational change from 6-s-trans to 6-s-cis, as figured in a retinal molecule, plays an important role in the formation of the retinochrome analog, based on the estimation of opsin shifts for retinal analogs locked in the 6-s conformation. Thus the conformation of the 6-7 single bond in the native retinochrome was suggested to be 6-s-cis. Analysis of the circular dichroic spectra of retinochrome analogs revealed that the 6-s conformation is independent of the appearance of the beta-band. The stereoselectivity in the photoisomerization of the retinal analogs by a retinochrome template depends on the hydrophobic binding in the region of the beta-ionone ring. PMID- 8234477 TI - Effects of bound monoclonal antibodies on the decay of the phototransformation intermediates I700(1,2) from native Avena phytochrome. AB - The kinetics of the microsecond phototransformation intermediates of 124 kDa Avena phytochrome (I700(1,2) were studied in the presence of bound monoclonal antibodies at various temperatures. A global analysis was applied to the decays at all wavelengths at each temperature in order to derive the rate constants and the decay-associated spectra of the three decay components. Monoclonal antibodies bound to specific epitopes altered the Arrhenius parameters of both I700(1,2) decay components. The strongest influence on these parameters was observed with OAT 8 (epitope between residues 624 and 686), which decreased by more than 50% the activation parameters of both components. This decrease is interpreted to result from an increased flexibility induced by this antibody in the ground state or in the transition state of bonds changing during the decay of both I700 transients. Thus, the OAT 8 epitope appears to be functionally important during the decay of the I700(1,2) intermediates. For the case of I700(1), bound OAT 23 and OAT 25 (epitopes between residues 1 and 66) reduced even further the relatively small flexibility of these bonds in the red light-absorbing form of phytochrome (Pr) without antibodies, as reflected by the high preexponential factors for its decay. This resulted also in higher activation energies for this decay in the presence of the antibodies. Thus, the amino-terminus should act as a rigid spacer of the chromophore cavity without affecting it during the microsecond transformation, because the Arrhenius parameters for these decays are similar to those for small phytochrome. The possible implications of the influence of the various antibodies on the bleaching remaining after the decay of I700(1,2) are discussed. PMID- 8234478 TI - Action spectrum for subliminal light control of adaptation in Phycomyces phototropism. AB - Adaptation processes enable phototropism and other blue light responses of Phycomyces to operate over a 10-decade range of fluence rate. Phototropic latency, used routinely to monitor the kinetics of sensitivity recovery after a step down in fluence rate, can be shortened by application of dim light for 35 min during the early part of the latency period. This light is termed subliminal, because it does not elicit phototropism under these experimental conditions; rather, it exerts its influence on the underlying adaptation kinetics. Fluence rate-response data for this latency reduction, obtained at 17 wavelengths of subliminal light from 347 to 742 nm, showed a variety of shapes that could be fit by zero, one, or two sigmoidal components, plus a constant term. At most wavelengths, the fluence-rate threshold for latency reduction by subliminal light tended to be well below the absolute threshold for phototropism, indicating that this effect is highly sensitive. An action spectrum for the sensitivity of the subliminal light effect, derived from the fluence rate-response curves, shows major peaks around 400 and 500 nm and a broad band from 570 to 670 nm, followed by a steep absorption edge. The sensitivity in the near ultraviolet region is relatively very low. The magnitude of the latency reduction also depends strongly on wavelength with a maximum at about 450 nm. The fluence-rate response data and the action spectrum--which is markedly different from that for phototropism and other blue-light responses of Phycomyces--indicate the participation of multiple pigments, or pigment states, in the photocontrol of adaptation. PMID- 8234479 TI - Multifrequency cross-correlation phase fluorometry of chlorophyll a fluorescence in thylakoid and PSII-enriched membranes. AB - We present here a comparative study on the decay of chlorophyll (Chl) a fluorescence yield in thylakoid membranes and photosystem II (PSII)-enriched samples, measured with multifrequency cross-correlation phase fluorometry. These measurements confirm the general conclusions of Van Mieghem et al. (Biochim. Biophys. Acta 1100, 198-206, 1992), obtained with a flash method, on the effects of reduction of the primary quinone acceptor (QA) on Chl a fluorescence yield of PSII. Different states of the reaction centers of PSII were produced by: (1) pretreatment with sodium dithionite and methyl viologen followed by laser illumination: the doubly reduced QA (QAH2) centers; (2) with laser illumination or pretreatment with diuron: QA- centers; and (3) the addition of micromolar concentration of dichlorobenzoquinone (DCBQ): oxidized QA centers. The data were analyzed with Lorentzian distribution as well as with multiexponential fluorescence decay functions. The analysis with Lorentzian distribution function showed that upon formation of QA-, the major lifetime distribution peak shifted to longer lifetimes: from 0.25 ns to 1.66 ns (pea thylakoid membranes) and from 0.24 ns to 1.31 ns (core PSII). However, when QAH2 was formed, the lifetime distribution peaks shifted back to shorter lifetimes (0.57-0.77 ns) both in thylakoids and PSII membranes. Multiexponential analysis showed three lifetime components: fast (40-400 ps), middle (300-1500 ps) and slow (5-25 ns). When QA- was formed in PSII centers, the amplitude of the fast component decreased, but both the amplitude and the lifetime of the middle component increased severalfold.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234480 TI - Light emission accompanies oxygen uptake during the peroxidative metabolism of tetracyclines. AB - Tetracycline molecules offer several sites for peroxidative metabolism of the type known to lead to oxygen consumption and electronic excitation. Accordingly, when tetracycline and chlortetracycline were exposed to horseradish peroxidase in the presence of hydrogen peroxide, oxygen was taken up and light emission was observed. The overall quantum yield of chemiluminescence is on the order of 10( 6), but that of chemiexcitation may be orders of magnitude higher as suggested by studies of sensitized emission. Given the widespread distribution of peroxidases, the formation of highly reactive metabolites of tetracycline may have biological importance. PMID- 8234481 TI - Frequency of ultraviolet radiation-induced mutation at the hprt locus in repair proficient murine fibroblasts transfected with the denV gene of bacteriophage T4. AB - The frequency of spontaneous and ultraviolet radiation (UVR)-induced mutation at the hprt locus was determined in control and denV-transfected, repair-proficient murine fibroblasts. Control cells removed an average of 25% of pyrimidine dimers induced by exposure to 150 J/m2 UVR from an FS40 sunlamp within 24 h; under the same conditions of induction and repair, denV-transfected cells removed an average of 71% of pyrimidine dimers. Control cells were somewhat more resistant than denV-transfected cells to killing by UVR. The average frequency of spontaneous mutation at the hprt locus for control and denV-transfected cells was 3 and 15 6-thioguanine (6-TG)-resistant colonies per 10(6) surviving cells, respectively; there was no statistically significant difference between control and denV-transfected cells. However, after exposure to 75 or 150 J/m2 UVR, denV transfected cells had a significantly lower frequency of mutation to 6-TG resistance. After exposure to a fluence of 75 J/m2, the average frequency of UVR induced mutation at the hprt locus was 166 mutant colonies per 10(6) surviving cells for control cells and 92 mutant colonies for denV-transfected cells; after 150 J/m2, control cells had 205 6-TG-resistant colonies per 10(6) cells, while denV-transfected cells had 61 mutant colonies. These results demonstrate that UVR induced pyrimidine dimers are mutagenic photoproducts in mammalian cells. PMID- 8234482 TI - DNA repair in the variable platyfish (Xiphophorus variatus) irradiated in vivo with ultraviolet B light. AB - Dark- and light-dependent DNA repair processes were studied in vivo in the variable platyfish, Xiphophorus variatus. Excision (dark) repair of the (6-4) photoproduct was more efficient than that of the cyclobutane dimer with approximately 70% of the (6-4) photoproducts removed by 24 h post-UVB radiation compared to approximately 30% of the cyclobutane dimers. Exposure to photoreactivating light resulted in rapid loss of most (> 90%) of the cyclobutane dimers and increased excision repair of the (6-4) photoproduct. Preexposure to photoreactivating light 8 h prior to UVB radiation increased the rate of photoreactivation two-fold. PMID- 8234483 TI - Small circular plasmids of the eukaryote Dictyostelium purpureum define two novel plasmid families. AB - Two novel groups of circular, nuclear plasmids were discovered in the simple eukaryote Dictyostelium purpureum. They define two new Dictyostelium plasmid families each containing three members: Dpp1A, Dpp1B, and Dpp2 in the Dpp1 family, and Dpp3, Dpp4, and Dpp5 in the Dpp3 family. These plasmids are among the smallest known, ranging in size from 1309 bp (Dpp1A and Dpp1B) to 1961 bp (Dpp4). Family members are very similar. The most distantly related members of the Dpp1 family (Dpp1A and Dpp2) are 89% identical, while the most distantly related members of the Dpp3 family (Dpp3 and Dpp4) are 91% identical. No sequence similarity is found between these plasmid families, or to any other known plasmid or chromosomal DNA sequence. A 72-bp inverted repeat present in one copy in Dpp1A and Dpp1B is tandemly repeated in Dpp2. The Dpp3 family contains a region of 102 160 nucleotides rich in short, overlapping direct sequence repeats. This region is present once in Dpp3 and Dpp5 and is tandemly repeated in Dpp4. The repeat structures in the Dpp1 and Dpp3 families are relatively rich in GC base pairs (29 41%) in comparison to the unique sequence regions of the plasmids (16-22%). The longest open reading frame (ORF) beginning with an AUG codon in these plasmids is 168 bp in the Dpp3 family, although longer ORFs without AUG start codons (up to 201 bp) also exist. Northern blot analysis did not detect any plasmid-specific transcripts in total RNA prepared from vegetative cells carrying Dpp1A or Dpp3. PMID- 8234484 TI - Fine mapping and DNA sequence of replication functions of Bacillus thuringiensis plasmid pTX14-3. AB - pTX14-3 is a 7.5-kb cryptic plasmid isolated from a Bacillus thuringiensis subspecies israelensis strain. Like many other small plasmids in gram-positive bacteria, pTX14-3 replicates via a single-stranded DNA intermediate. The nucleotide sequence of the replication region was determined and an open reading frame of 636 base pairs encoding a protein necessary for plasmid replication was identified by deletion analysis. No significant homology was found between this open reading frame and those encoding replication proteins identified on other plasmids isolated from gram-positive bacteria, nor could we find any homology to plus origins from other single-stranded DNA plasmids. Consequently, it seems that the replicon of pTX14-3 belongs to a new family of replicons in the group of single-stranded DNA plasmids. The sequence of the single-strand origin (i.e., the minus origin) responsible for the conversion of single-stranded plasmid DNA to double-stranded plasmid DNA was also determined. A partial homology between the minus origin of pTX14-3 and the Bacillus subtilis plasmid pBAA1 was identified. A previously identified locus that suppresses formation of high molecular weight multimers was also minimized and sequenced. PMID- 8234485 TI - Relationship between the replication functions of Streptomyces plasmids pJV1 and pIJ101. AB - The essential replication region of the Streptomyces phaeochromogenes plasmid pJV1 was sequenced and compared to the equivalent region of the well characterized Streptomyces plasmid pIJ101. The sequence revealed a similar organization in both plasmids, including a conserved region within both plasmid origins, which is just upstream of a gene encoding the replication protein (Rep). The Rep proteins of pJV1 and pIJ101 are very similar, and both have a conserved motif, also present in Rep proteins of single-stranded DNA plasmids of the low G + C gram+ bacteria that includes a tyrosine residue presumably involved in creating the single-stranded nick that initiates the replication event. A co integration experiment between these two replicons led to the formation of a novel replicon with a hybrid origin and allowed the identification of the nick site where replication starts, also showing that the replication functions of both plasmids are related. PMID- 8234486 TI - IS231V and W from Bacillus thuringiensis subsp. israelensis, two distant members of the IS231 family of insertion sequences. AB - IS231 constitutes a family of related insertion sequences (IS) from Bacillus thuringiensis. Two new IS231-related elements, IS231V and IS231W, have been isolated from the 72-MDa plasmid of B. thuringiensis subsp. israelensis. These closely related 1964-bp IS are delimited by 22-bp imperfect inverted repeats strongly similar to those of the other iso-IS231. Although the other known IS231 harbor a single long open reading frame (ORF), IS231V and W display two slightly overlapping ORF on the same DNA strand. They show about 50% identity with the transposase of the other iso-IS231. A frameshifting model is proposed for the synthesis of a fusion product which would constitute their active transposase. PMID- 8234487 TI - A transformation vector for dictyostelium discoideum with a new selectable marker bsr. AB - A new selectable marker for transformation of Dictyostelium discoideum cells was constructed by using the bsr gene from Bacillus cereus, which confers resistance to Blasticidin S. The bsr gene was driven by Dictyostelium actin 15 promoter and Dictyostelium actin 8 terminator for expression in Dictyostelium cells. To demonstrate the feasibility of using the bsr marker, we constructed an extrachromosomal replication vector by replacing the Neor gene of pnDeI (B. Leiting and A. Noegel (1988) Plasmid 20, 241-248) with the bsr gene cassette. A mutant Dictyostelium actin 15 gene was constructed and inserted into the vector. Dictyostelium cells were transformed with the resulting vector and then transformants were selected with Blasticidin S. The selected cells showed high level expression of the mutant actin, indicating an efficient selection of transformed cells with the bsr marker. PMID- 8234488 TI - Target sequence specificity of transposon Tn5 in the absence of major hotspots in the plasmid pBR322: identification of a new hotspot. AB - The plasmid pLB11-1 is a pBR322 derivative in which part of the tetracycline resistance (tet) gene (basepair coordinates 23 to 375), containing five hotspots for Tn5 insertion (D. E. Berg et al., Genetics 105, 813-828, 1983), has been replaced with a 5.1-kb fragment of Escherichia coli genomic DNA encoding an osmoregulatory function. Restriction mapping of 40 pLB11-1::Tn5 derivatives, chosen at random from 240 Tn5 insertion derivatives with an unaltered osmotolerant phenotype, placed Tn5 in the vector portion of 20 clones. The majority of these insertions (16/20) were located in a 0.09-kb region immediately downstream of the beta-lactamase (bla) gene. Nucleotide (nt) sequence analysis of seven insertions from this group revealed an identical site of insertion within pBR322, representing a hitherto unidentified hotspot for Tn5 insertion. The target sequence, 5'GTCTGACGC, was found to be duplicated in these cases. PMID- 8234489 TI - Effects of plasmid pBR322 on respiratory and ATPase activities in Escherichia coli. AB - The effects of antibiotic resistance plasmids on respiratory and ATPase activities were investigated in Escherichia coli. The rates of oxygen consumption coupled to the oxidation of succinate and NADH, ATP hydrolysis, and ATP-Pi exchange were measured in isolated membrane vesicles prepared from HB101 strains that contained derivatives of plasmid pBR322. The rates of oxygen consumption coupled to the oxidation of NADH were independent of the presence or absence of plasmids in the strains from which the vesicles were prepared. In contrast, the mean rates of oxygen consumption coupled to the oxidation of succinate, ATP hydrolysis, and ATP-Pi exchange were 140-292% higher in vesicles made from plasmid-containing strains than from plasmid-free HB101 and independent of the expression of the plasmid-encoded tetracycline/H+ antiporter. PMID- 8234490 TI - The tet(K) gene of plasmid pT181 of Staphylococcus aureus encodes an efflux protein that contains 14 transmembrane helices. AB - The corrected DNA sequence of the tet(K) structural gene, encoding the tetracycline efflux protein from Staphylococcus aureus, is reported. The Tet(K) protein is homologous to related tetracycline efflux proteins throughout the protein, including at the C-terminal end. Hydropathy plotting now clearly indicates that the Tet(K) protein contains 14 transmembrane alpha-helices. PMID- 8234491 TI - Insertion of transposon Tn5seq1 into G+C-rich DNA of Streptomyces avermitilis: generation of 8-, 9-, and 10-bp duplications. AB - The sites for 24 Tn5seq1 insertions into G+C-rich DNA of Streptomyces avermitilis were determined. Although Tn5-based transposons usually generate a 9-bp duplication at their insertion site, one Tn5seq1 insertion generated a 10-bp duplication and another insertion an 8-bp duplication. PMID- 8234492 TI - Transformation of Mycoplasma pneumoniae with Tn4001 by electroporation. AB - Mycoplasma pneumoniae was transformed with the Staphylococcus aureus transposon Tn4001 by electroporation. A transformation frequency of 10(-3) to 10(-5)/colony forming unit was observed using 30.0 microgram plasmid DNA and 10(7)-10(8) M. pneumoniae colony-forming units. DNA hybridization analyses using standard and pulsed field agarose gel electrophoresis confirmed chromosomal insertion of the transposon, apparently by a transpositional mechanism into random sites. These studies demonstrate the functionality of Tn4001 in M. pneumoniae and suggest its potential as a genetic tool in this mycoplasma. PMID- 8234493 TI - The genetics and basic biology of Mycoplasma pulmonis: how much is actually Acholeplasma? PMID- 8234494 TI - The OCT plasmid encodes D-lysine membrane transport and catabolic enzymes in Pseudomonas putida. AB - Pseudomonas putida (oleovorans) (Pp(OCT)) cured of its OCT plasmid (Pp) no longer grows on D-lysine. Conjugation of PpTrp- with three different methionine auxotrophs carrying the OCT plasmid resulted in PpTrp- (OCT) organisms that grew on D-lysine. Three early D-lysine catabolic enzymes encoded by the OCT plasmid are a lysine racemase, the proposed conversion of D-lysine to delta 1-piperidine 2-carboxylate (P2C), for which we provide evidence, and P2C reductase which converts P2C to pipecolate. In Pp the rate of active D-lysine transport is reduced by 30 to 50%. We consider this to reflect a loss of the gene for the D lysine carrier while the remaining activity is due to a chromosomally coded L lysine or D-ornithine carrier or both. The membrane carriers and catabolic enzymes for D- and L-lysine in P. putida P2 and for L-lysine in Pp(OCT) are chromosomally coded. PMID- 8234495 TI - Construction of shuttle plasmids which can be efficiently mobilized from Escherichia coli into the chromatically adapting cyanobacterium, Fremyella diplosiphon. AB - In some strains of cyanobacteria the composition of the light-harvesting antennae is determined by the color of available light. The mechanism of this chromatic adaptation involves the regulation of gene expression by red and green light and has been most studied in Fremyella diplosiphon (Calothrix sp. PCC 7601), a filamentous cyanobacterium for which there has been no reported means of genetic manipulation. We have constructed shuttle plasmids which can be efficiently mobilized by RP4 from Escherichia coli into F. diplosiphon and which can be recovered from transconjugant F. diplosiphon and returned to E. coli by transformation. The ability of these plasmids to replicate in F. diplosiphon is conferred by an 8.0-kb DNA fragment isolated from pFDA, a plasmid native to F. diplosiphon. To create these shuttle plasmids the 8.0-kb fragment was cloned into pJCF22, a mobilizable plasmid constructed from oriV and bom from pBR322, cat from pACYC184 and aphA from pACYC177.pJCF22 lacks sites for the restriction enzymes FdiI and II. Transconjugant F. diplosiphon containing shuttle plasmid pJCF62 are resistant to chloramphenicol and highly resistant to the aminoglycosides, G418 and neomycin. When aadA from the omega interposon was incorporated into a shuttle plasmid transconjugant F. diplosiphon could also be selected with streptomycin or spectinomycin. In F. diplosiphon shuttle plasmid pJCF62 replicates with a minimum copy number of seven. The oriV for replication in F. diplosiphon was localized to a 2.8-kb region within the cyanobacterial part of pJCF62. The presence on a shuttle plasmid of a single recognition site for FdiI reduced the efficiency of mobilization into F. diplosiphon by 5- to 10-fold. Restriction at this site was prevented when the E. coli donor strain in the mating contained the enzyme Eco47II methylase. PMID- 8234496 TI - Biodegradation of the polyurethane foam covering of breast implants. AB - Although it is generally accepted that polyurethane-covered breast implants have decreased the incidence of clinical capsular contracture, there remain many unanswered questions regarding the physical and chemical degradation of the polyurethane foam covering itself. We have systematically studied the fibrous capsule and polyurethane foam recovered from human breast "explants" in an effort to characterize more precisely the biodegradation of polyurethane foam in the human body. Seventy-five freshly retrieved polyurethane-covered implants and surrounding capsule from 47 patients have been analyzed. Capsular tissue from several sampling sites around the surface of the implants was digested in a collagenase solution until foam was recovered or all tissue was digested. Additional samples were fixed in 10% formalin. Scanning electron microscopy was used to look for structural changes in the recovered intact foam and to determine the foam strut widths. Fourier transform IR spectroscopy and x-ray photoelectron spectroscopy were used to analyze the chemical composition of the polyurethane. The formalin-preserved capsule samples were examined histologically for further evidence of foam degradation. Of the 75 prostheses analyzed, 36 (48 percent) were removed because of capsular contracture and 10 (13 percent) because of infection or exposure of the prosthesis. The remaining 29 (39 percent) implants were removed for various other reasons. Visibly intact foam was recovered from 36 (48 percent) prostheses after enzymatic digestion of capsule tissue. There was a progressive decline in the ability to recover intact foam as the total implantation time increased. Scanning electron microscopy revealed fractures and fissures in the foam structure and thinning of the polyurethane struts. The mean strut width of control, unimplanted foam was 49 +/- 1.5 microns (+/- SEM). Retrieved foam from implants which developed capsular contracture and the infected implants had strut widths of 30 +/- 3.1 and 32 +/- 3.1 microns, respectively. In implants removed for other reasons, the polyurethane foam strut width was 41.2 +/- 2.3 microns. Despite an inability to recover visibly intact foam from 39 specimens, standard light microscopy of 37 of these same specimens showed residual polyurethane still present in the capsule. Various degrees of scalloping and fracturing of the foam were seen in the histologic sections. There is convincing evidence by scanning electron microscopy and histology that polyurethane is degrading. It was not possible to quantitate accurately the rate of degradation, but factors such as capsular contracture, infection, and time appear to have a role in the biodegradation of polyurethane in the human body. These relationships require further study. PMID- 8234497 TI - Sagittal synostosis: quantitative assessment of presenting deformity and surgical results based on CT scans. AB - We reviewed our experience with nine consecutive patients with untreated isolated nonsyndromic sagittal synostosis. Using a method of 14 clinically relevant measurements taken from preoperative and postoperative CT scan images of these patients, we documented their presenting skeletal dysmorphology and the results of surgical correction at least 1 year after operation. Significant preoperative findings included an elongated cranial vault length that averaged 103 percent of normal and a narrowed cranial vault width both anteriorly at 92 percent and posteriorly at 86 percent of normal. Results of surgical correction, as documented by CT scan measurements, included normalization of the cranial length to 100 percent and of the anterior width to 101 percent of normal and improvement (but undercorrection) of the posterior width to 94 percent of normal. Quantitative measurement of CT scan images confirmed clinically observed findings in these patients before suture release and reconstruction and proved useful in assessing the surgical results achieved. PMID- 8234498 TI - Crouzon syndrome: quantitative assessment of presenting deformity and surgical results based on CT scans. AB - We reviewed our experience with 14 children who presented sequentially with untreated Crouzon syndrome and whose cranial vault presentation was with bilateral coronal synostosis. Using a method of 14 measurements in the cranio orbitozygomatic region taken from preoperative and postoperative CT scans in these patients, we documented their presenting skeletal morphology and the results of surgical correction at least 1 year after operation. Our preoperative measurements confirmed a widened anterior cranial vault at 108 percent of normal and a cranial length averaging only 92 percent of normal. In comparison with age matched controls, orbital measurements revealed a widened anterior interorbital distance at 122 percent of normal, an increased intertemporal width at 121 percent of normal, globe protrusion at 119 percent of normal, and a short medial orbital wall distance at only 86 percent of normal. The distance between the zygomatic buttresses and the interarch distance were found to be increased at 106 and 103 percent of normal, respectively. The zygomatic arch lengths were substantially shortened at only 87 percent of age-matched control values. These findings confirmed clinical observations of brachycephalic anterior cranial vaults with shallow, hyperteloric orbits and globe proptosis. Generally, in these patients the midface is horizontally retrusive and transversely wide, reflected in wide and shortened zygomas. Assessment of the postoperative results at least 1 year later showed no significant changes in any craniofacial measurements. Our findings indicate that early surgical attempts to decompress and reshape the cranio-orbital regions may limit the effects of increased intracranial pressure but do not correct the deformity as judged by CT scan evaluation at least 1 year later. Over the period of the study, the Crouzon deformity did not worsen after surgery, but the measurements remained far from normal. PMID- 8234499 TI - The mask rhinoplasty: a technique for the treatment of Binder's syndrome and related disorders. AB - Congenital and acquired facial deformities in which the nose is small relative to the rest of the face present a problem in reconstruction. Binder's syndrome is the most common of such deformities, where the noise is both short and lacking tip projection. A method of reconstructing such deformities is described. By means of a coronal skin incision and an approach through the upper buccal sulcus, the nasal soft tissues including the alar cartilages are mobilized. Nasal lengthening and improved tip projection are achieved and supported by a cantilevered graft of lyocartilage. The defect created within the nasal vestibule is repaired by means of mucosal flaps raised from the upper buccal sulcus. Ten cases have so far been treated successfully using this technique we call mask rhinoplasty. PMID- 8234500 TI - Fetal cleft lip and palate: sonographic diagnosis and postnatal outcome. AB - This is a retrospective analysis of neonatal outcome in 32 fetuses diagnosed over a 3 1/2-year period as having cleft lip and palate by ultrasonographic examination. Unilateral versus bilateral complete cleft lip was accurately diagnosed in fetuses after 16 weeks' gestation. Incomplete cleft lip was more easily detected after 27 weeks' gestation. In only three newborns was the severity of the cleft overappraised. In no instance was a complete cleft lip mistakenly diagnosed as an incomplete form. The fetuses with labial clefting also were categorized sonographically as those without associated abnormalities (n = 15) and those with other detectable anomalies (n = 17), of which 5 had autosomal trisomy. One neonate assessed in utero to have an isolated unilateral incomplete cleft lip had vertebral/rib anomalies and died soon after birth of unsuspected pulmonic stenosis. This study confirms the accuracy of prenatal sonographic examination and underscores the high incidence of spontaneous fetal loss (terathanasia) that occurs whenever cleft lip/palate is associated with aneuploidy or other malformations. PMID- 8234501 TI - Expanded unilateral forehead flap (sail flap) for coverage of opposite forehead defect. AB - This report details our experience with five patients who had skin defects of their forehead that were covered with an expanded unilateral forehead flap from the contralateral side that we call the sail flap. Skin expansion provides an ideal thin flap for the forehead aesthetic unit. The viability of the sail flap is sufficient through expansion, and it is available in suitable size and shape for the defect. The donor site is minimal and well accepted because the scar coincides with the hairline. PMID- 8234502 TI - Gore-Tex soft-tissue patch frontalis suspension technique in congenital ptosis and in blepharophimosis-ptosis syndrome. AB - The frontalis suspension technique is the surgical method of choice in ptosis patients with a levator function of 2 mm or less. This type of ptosis is found unilaterally or bilaterally in isolated congenital ptosis and mostly bilaterally in blepharophimosis-ptosis patients. The frontalis suspension technique is frequently used in children being operated on because of congenital ptosis, where the excision of the fasciae latae can be avoided so as not to risk scarring the donor area. Since the new heterologous material polytetrafluoroethylene (Gore-Tex soft-tissue patch) has been available (1986), it has been used on our ptosis patients with minimal levator function instead of the formerly much used fasciae latae. Thirty-seven frontalis suspension procedures have been performed in 26 patients, of whom 6 were children who suffered from bilateral blepharophimosis ptosis syndrome. All ptosis procedures were performed as Fox pentagons, and in the combined cases, the medial canthoplasty was carried out as a bilateral one stage procedure. The postoperative healing was free of complications. Three years postoperatively one patient did exhibit rejection of one of the two implanted Gore-Tex slings. In all other patients, the functional and cosmetic results were satisfying, and high biocompatibility of the Gore-Tex soft-tissue patch material could be shown. PMID- 8234503 TI - Eye socket reconstruction with free-flap transfer. AB - Patients afflicted with retinoblastoma or other malignant orbital tumors often undergo eyeball excision and irradiation in infancy, which results in severe deformities of the orbit. It is very difficult to achieve the reconstruction of such severely depressed and contracted eye sockets because both augmentation and formation of an eye socket are required. We usually perform a free-flap transfer with microvascular anastomoses in such situations and have experienced 25 cases since 1980. Various kinds of flaps, such as the groin flap, dorsalis pedis flap, scapular flap, and others, were utilized. The degree of depression and whether sufficient conjunctiva for lining of the eye socket remains are important in the selection and design of the donor flap. To obtain an orbit with a natural appearance, many of the patients required subsequent minor revision operations such as debulking, dermal fat grafts, canthoplasty, fascia suspension, and so on. PMID- 8234504 TI - The correction of lower lid malposition following lower lid blepharoplasty. AB - Lower eyelid malposition is the most common complication following lower eyelid blepharoplasty. This may take the form of a mild scleral show with the round eye syndrome or may progress to frank ectropion in some patients. When this problem occurs, it is imperative to soothe an already distressed patient by correcting the malposition by the most efficient method. A horizontal tightening of the lower lid at the lateral canthus can be helpful in many patients; however, in the more severe cases or in those in which the medial component is prominent, this procedure may not give the best correction. It is preferable to avoid using autogenous grafts in these patients because in most cases they will induce another incision line with potential deformity and, more important, are perceived by these sensitive patients as being more of an involved procedure. The preserved fasciae latae sling is an alternative procedure which is effective in the more severe cases of postblepharoplasty lower eyelid malposition, particularly in those cases in which the medial component of malposition is prominent. PMID- 8234505 TI - Intracapsular injection of triamcinolone for prevention of contracture. AB - The use of steroids for the prevention of implant capsular contracture is an attractive concept, but the method has been handicapped by difficulties getting the right amount of drug to the capsule at the right time. Triamcinolone "in the pocket" was found to be ineffective for prevention of capsular contracture. When the drug was injected into the capsule at 4 and 8 weeks postoperatively, it was very effective. The method for percutaneous injection of intracapsular steroid is explained in detail. PMID- 8234506 TI - "Mammacompliance": an objective technique for measuring capsular fibrosis. AB - The results achieved with silicone implants are usually assessed on the basis of the clinical and therefore subjective technique of Baker's classification. Yet, for scientific studies, more objective measurements are clearly necessary. For this reason, the calliper originally developed by Burkhardt and used for measuring compressibility of the breast was adapted, adding a force transducer and a distance-measuring device, which allowed us to plot a force-distance graph for each breast. In soft breasts, the curves obtained rise very slightly, while firm breasts produce sharply rising curves. The data computed from the graphs give the degree of "mammacompliance" and are expressed in centimeters at a defined force value. Sixty-eight augmented and reconstructed breasts were examined with this device. Measurements also were taken on a series of 59 normal, untreated breasts for comparison. A strong correlation was found between the mammacompliance values and Baker's classification. At the same time, the results revealed a number of clinical misjudgments. Our technique permits the objective assessment of capsular contracture. Being highly sensitive, it is an excellent method to control and monitor postoperative development and therapeutic concept. PMID- 8234507 TI - Tumescent technique for local anesthesia improves safety in large-volume liposuction. AB - The tumescent technique for local anesthesia improves the safety of large-volume liposuction ( > or = 1500 ml of fat) by virtually eliminating surgical blood loss and by completely eliminating the risks of general anesthesia. Results of two prospective studies of large-volume liposuction using the tumescent technique are reported. In 112 patients, the mean lidocaine dosage was 33.3 mg/kg, the mean volume of aspirated material was 2657 ml, and the mean volume of supernatant fat was 1945 ml. The mean volume of whole blood aspirated by liposuction was 18.5 ml. For each 1000 ml of fat removed, 9.7 ml of whole blood was suctioned. In 31 large volume liposuction patients treated in 1991, the mean difference between preoperative and 1-week postoperative hematocrits was -1.9 percent. The last 87 patients received no parenteral sedation. In a second study, a 75-kg woman received 35 mg/kg of lidocaine on two separate occasions, first without liposuction and 25 days later with liposuction; peak plasma lidocaine concentrations occurred at 14 and 11 hours after beginning the infiltration and were 2.37 and 1.86 micrograms/ml, respectively. PMID- 8234508 TI - Suction lipectomy: outcome relates to region-specific lipoprotein lipase activity and interval weight change. AB - To determine the effects of suction lipectomy on regional adipose tissue metabolism, nine women had repetitive circumferential measurements and biopsies of subcutaneous adipose tissue from a lipectomy site (site A) and a nonlipectomy site (site B) up to 12 months following lipectomy. Maximum reductions from preoperative baseline in weight, body mass index, and circumferences of sites A and B occurred at 3 months. Because of variable long-term compliance (6 to 12 months), we created a "last visit" time-point to assess adequately the effects of lipectomy for each individual. Not all subjects maintained reduction in site circumferences from 3 months to the last visit. The change in circumference of site A for that period was highly correlated with the change in circumference of site B (r = 0.828, p = 0.005). The change in circumference of site B, but not site A, at 3 months was related to the weight change above the weight of adipose tissue removed at suction lipectomy. Five subjects who were "sustained responders" to the lipectomy procedure were able to maintain or decrease circumferences of sites A and B from 3 months to the last visit. In contrast, four "limited responders" actually increased circumference of site A and had either no change or increased circumference of site B from 3 months to the last visit. The changes in circumference in both sites between 3 months and the last visit related to changes in body weight over the same interval. Ultimately, the decrement in circumference of site A was 3.4 percent (-2.4 cm) greater than that of site B (p = 0.0001). The response to lipectomy in site B, but not site A, between 3 months and the last visit was related to the change in fasting adipose tissue lipoprotein lipase from baseline to 3 months (r = 0.728, p = 0.026). This change in lipase activity in the control region may represent a metabolic defense of body weight in response to adipose tissue removal in the lipectomy site. PMID- 8234509 TI - Lower body lift with superficial fascial system suspension. AB - Multiple body contour deformities of the trunk and thighs are commonly treated in separate stages to limit postoperative complications and disability. Recent advances in the surgical design of the medial thigh lift and the lateral thigh/buttock lift along with an understanding of the functional anatomy of the superficial fascial system have significantly improved results and decreased complications of trunk/thigh lifts. The enhanced safety of current trunk/thigh lifts has allowed new combinations to treat multiple body contour deformities in a single stage. Laxity of the entire lower trunk and thigh regions can be treated in one stage in selected patients. The lower body lift combines the transverse flank/thigh/buttock lift and the fascial anchoring medial thigh lift in one operation. In addition to the expected tightening of the flank, buttocks, and total thighs, this procedure results in a surprising degree of epigastric and hypogastric tightening of mild to moderate abdominal laxity without direct surgical undermining or umbilical transposition. Ten patients having the lower body lift alone or in combination with liposuction and other body contouring procedures were followed for 6 to 24 months. The primary indication for surgery is moderate to severe soft-tissue laxity of the lower trunk and thighs with minimal or mild residual fat deposits. Skin contour irregularities due to skin laxity (cellulite of laxity) or to postliposuction adhesions are frequently present and may be severe. Patients with significant fat deposits may be treated initially with liposuction 3 to 4 months earlier to become candidates for this procedure. Key technical elements of this procedure include (1) both supine and lateral decubitus positioning with the hip flexed and abducted to allow overcorrection, (2) appropriate direct surgical undermining through superficial fascial system zones of adherence in the superior thigh while avoiding the lymphatics of the femoral triangle, (3) more distal undermining of the thighs with an atraumatic undermining cannula, (4) resection of redundant lower trunk and thigh tissues with incisional closure within high-cut bikini lines, and (5) complete superficial fascial system and dermal suspension throughout the repair. The lower body lift requires 5 to 6 hours of operating time, 2 units of autologous blood, 2 nights of nursing care, and 3 weeks off work. Although no major complications occurred, minor complications were significantly higher than with the component procedures alone and occurred in 50 percent of patients. Technical changes and improved patient selection have significantly improved the rate of complications in subsequent patients. Results have been consistent, and patient satisfaction has remained high. PMID- 8234510 TI - Comparison of rapid versus slow tissue expansion on skin-flap viability. AB - The effect of rapid versus slow tissue expansion on random-pattern skin-flap survival lengths was evaluated in two groups of pigs. Each group (group 1--slow; group 2--rapid) was further evaluated for the effect that delay (A), delay plus expansion (B), expansion only (C), and acute flap elevation (D) had on flap length viability. Intergroup and intragroup comparisons were made. In the slowly expanded group, significantly greater flap length viabilities were demonstrated in the slow expansion flaps as compared with the delay plus expander flaps and the delayed flaps, respectively. In the rapidly expanded group, the flap length viabilities of flaps C, B, and A were statistically equal. However, the flap length viability of the rapidly expanded flaps was statistically equal to that of the slowly expanded flaps (C1 = C2). The effect tissue expansion has on improving flap length viability as compared with the delay phenomenon may be related to the combination of a delay effect plus increased nutrient flow density that slow expansion produces. This combined metabolic enhancement is preserved with rapid tissue expansion. PMID- 8234511 TI - Clinical experience with the reverse lateral arm flap in soft-tissue coverage of the elbow. AB - Coverage of soft-tissue defects of the posterior aspect of the elbow is a difficult problem. Many techniques have been published, mostly case reports. The radial recurrent fasciocutaneous flap as described by Maruyama and Takeuchi represents one of the best solutions for medium-sized defects. We used it successfully in five clinical cases. The anatomy of this flap, based on the radial recurrent artery, has been confirmed by our cadaver studies, and a technique to raise this flap safely will be described. PMID- 8234512 TI - Anatomic basis of local muscle flaps in the distal third of the leg. AB - Soft-tissue defects of the distal third of the tibia are considered to be the realm of free-tissue transfer. We have found clinically that several local muscles can be used reliably in this area. The purposes of this study were: (1) to evaluate the potential use of the local muscles for soft-tissue coverage in the lower third of the leg and (2) to obtain numerical data that could be used preoperatively in the selection of potential local muscle flaps. All potential muscles, excluding the gastrocnemius, plantaris, and popliteus, from 10 fresh frozen legs were examined and the following details recorded: (1) the distance above the medial malleolus that the muscle bellies ended (musculotendinous junction), (2) the distance between the medial malleolus and the distal end of the transposed flaps (reach), (3) the area of tibia that could be covered, and (4) the vascular supply to these muscles. The soleus, extensor digitorum longus and peroneus tertius, extensor hallucis longus, peroneus brevis, and flexor digitorum longus were found to be the most anatomically suited muscles for local transposition to selected lower-third defects. We have worked with these muscles clinically and have found them to be useful and reliable when chosen appropriately. PMID- 8234513 TI - Fetal tendon healing: development of an experimental model. AB - An experimental model was developed to study the process of fetal tendon healing. The flexor digitorum profundus tendons of the right hindlimb of 14 fetal lambs were partially lacerated at 100 days' gestation (term 145 days) and then studied macroscopically and histologically at several postinjury intervals (2, 4, 7, 14, 28, 42, and 56 days); two lambs were studied at each interval. A similar procedure was done in 14 adult sheep, who served as a control group. The fetal lambs showed no subcutaneous scarring, the digital sheath and tendon healed 2 weeks after injury, and a smooth, gliding surface was reconstituted. Collagen fibers were randomly arranged at 1 week but became organized along the tendon axis by 2 weeks. No adhesions, ruptures, or triggering was noted in the healing fetal tendons. Normal morphology was restored by 6 weeks. In the adult, dense subcutaneous scarring was noted, the digital sheath healed by 4 weeks and the tendon gap by 6 weeks, but a smooth gliding surface was not restored. Collagen fibers were randomly arranged at 2 weeks and became organized along the tendon axis by 4 weeks after injury. There were no dense adhesions or ruptures, but 25 percent of the tendons showed triggering. PMID- 8234514 TI - Assessment of ischemia-induced reperfusion injury in the pig latissimus dorsi myocutaneous flap model. AB - Experiments were conducted to assess ischemia-induced reperfusion injury in the pig latissimus dorsi myocutaneous flap model. Forty Yorkshire pigs (19.5 +/- 0.6 kg) were assigned to groups A, B, C, and D (n = 10 pigs). Bilateral 8 x 13 cm latissimus dorsi myocutaneous flaps were constructed in each pig, and one flap was assigned to ischemic treatment and the contralateral flap served as a nonischemic control. The treatment flaps in groups A, B, C, and D were subjected to 2, 4, 6, and 8 hours of warm global ischemia, respectively. Pigs in groups A, B, C, and D were divided into two subgroups (n = 5 pigs), and extents of skin and muscle necrosis in control and treatment flaps were assessed with the fluorescein and nitroblue tetrazolium dye stain tests, respectively, after 2 and 7 days of reperfusion. Significantly (p < 0.01) greater extents of skin and muscle necrosis were observed in latissimus dorsi myocutaneous flaps subjected to 4, 6, or 8 hours of ischemia compared with their contralateral controls. Extents of skin and muscle necrosis also increased significantly (p < 0.01) with increases in ischemia time in treatment flaps. Of particular importance was the observation that there was no significant difference in the extent of skin or muscle necrosis between 2 and 7 days of reperfusion in all control and treatment groups. This observation indicates that 2 days of reperfusion time is adequate to assess the maximum extent of skin and muscle ischemia-induced reperfusion injury in pig latissimus dorsi myocutaneous flaps. Furthermore, it was observed that 1-cm segments of latissimus dorsi muscle were not too thick to allow the use of the nitroblue tetrazolium dye stain test for assessment of muscle viability, as judged by the highly correlated (r = 0.98, n = 40) linear relationship between assessment of muscle viability from one transverse cut surface of muscle segments and by weighing total viable and nonviable muscles dissected from the flaps according to the nitroblue tetrazolium dye stain on both transverse cut surfaces. It is important to note that the maximum length of the latissimus dorsi myocutaneous flap model for ischemia-induced reperfusion injury research should not exceed the maximum length of skin viability in the nonischemic control in order to avoid the complication of skin necrosis due to excessive length of skin.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234515 TI - Endoscopic diagnosis and treatment of persistent halitosis after pharyngeal flap surgery. AB - A patient presenting with persistent dramatically offensive fetid malodor 1 year after pharyngeal flap surgery is described. Nasendoscopic examination revealed the presence of a foreign body within the nasopharynx lying on the flap. The foreign body was immediately removed by endoscopic telebiopter and revealed itself to be a gauze pad forgotten during the pharyngeal flap surgery. Modern endoscopic equipment allowed both quick diagnosis of this very rare complication and immediate and simple treatment under topical anesthesia. PMID- 8234516 TI - Systemic inflammatory disorder related to fibrous breast capsules after silicone implant removal. AB - Silicone breast implants have been associated with connective-tissue inflammatory syndromes such as systemic sclerosis, and as with other artificial breast prostheses, fibrous capsules tend to form around the implants. The capsular tissue is generally considered inert and typically is left in situ when the prostheses are explanted. We report a patient who formed symptomatic bilateral submammary cysts associated with pain, swelling, arthralgia, fever, axillary lymphadenopathy, accelerated erythrocyte sedimentation rate, and antinuclear antibody following removal of intact silicone breast implants without capsulectomy. Clinical improvement followed removal of the capsules, which histologically displayed fragments of silicone, fibrous tissue, and inflammatory cells. Our experience suggests that when silicone breast implants are thought to be the cause of a clinical inflammatory syndrome characterized by mammary pain, swelling, arthralgia, or serologic abnormalities, consideration should be given to removing the capsules entirely so that the chance of a perpetuating reaction will be reduced. PMID- 8234517 TI - Modification of Mustarde technique for correction of epicanthus in Asian patients. AB - For correction of epicanthus in Asian patients, a modification of the Mustarde technique was done to avoid excessive scarring and to retain ethnic characteristics. The method includes adapting half Y-V advancement instead of transposing two cephalic flaps in the four-flap technique of Mustarde. Three patients with blepharophimosis and a patient with Down syndrome were operated on by this modification with satisfactory results. PMID- 8234518 TI - An approach to the senile upper lip. AB - Senile changes in the upper lip produce not only alterations in the length and shape of the vermilion border but also elongation of the skin area and vertical wrinkles. Correction of these changes can be obtained by means of the surgical technique described in this paper. The surgical technique must be exact and performed patiently. The complications are temporary and not serious. The results are satisfactory. PMID- 8234519 TI - An electronic device for surgical glove testing. AB - An electronic glove tester complying with safety regulations for electric current flow is described. In our laboratory, it was able to detect 100 percent of glove punctures caused by 4-0 and 6-0 surgical needles. When used regularly in the surgical suite, this device may help to minimize the surgeon's exposure to patients' blood and body fluids. PMID- 8234520 TI - Is a free or pedicled TRAM flap safe after liposuction? PMID- 8234521 TI - Some thoughts and questions about abdominal fat. PMID- 8234522 TI - Pregnancy after TRAM flap. PMID- 8234523 TI - Augmentation of craniofacial skeleton. PMID- 8234524 TI - The effect of denervation on soft-tissue infection. PMID- 8234525 TI - A new and exact way to make an osteotomy at the nasofrontal bone junction. PMID- 8234526 TI - Interplast program in Peshawar, Pakistan. PMID- 8234527 TI - Biological creep. PMID- 8234528 TI - The dorsal skin-flap model in the rat. PMID- 8234529 TI - Three-dimensional imaging and computer-generated models in complex facial reconstructions. PMID- 8234530 TI - Thoughts of a reader from faraway Russia about your journal--joys and problems. PMID- 8234531 TI - Le Temps. PMID- 8234532 TI - Giant pilar tumor of the scalp. PMID- 8234533 TI - [Changes in monoamine and monoamine metabolites in the nucleus accumbens and locomotor activity caused by p-chloroamphetamine: studies with intracerebral dialysis in freely moving rats]. AB - The effect of i. p. administration of p-chloroamphetamine (PCA) (2mg/kg) on the dopamine (DA), serotonin (5-HT), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) was estimated in freely moving rats by intracerebral dialysis in the nucleus accumbens. Locomotor activity was measured simultaneously by the Animex. After PCA administration, locomotor activity increased significantly in hours 1-2. DA concentration was elevated statistically for 1-2 hours after PCA treatment. And the change of locomotor activity and DA levels were correlated. 5-HT was slightly elevated for 1-2 hours after PCA treatment, although this was not statistical significance through the whole experimental period. These results suggest that an increase in locomotor activity induced by PCA may be explained by activation of the dopaminergic systems in the nucleus accumbens. DOPAC, HVA and 5-HIAA were suppressed significantly during 1-6th. These results suggest that suppression of DOPAC, HVA and 5-HIAA induced by PCA may be explained by PCA's pharmacological effect such as inhibition of monoamineoxidase. PMID- 8234534 TI - [Emotional condition in alcohol withdrawal acute psychosis]. AB - It is widely known that the acute psychotic states in alcohol withdrawal period, especially delirium tremens and acute hallucinosis, are often accompanied by intense anxiety and fear. On the other hand it is also noted that the emotional condition in delirium tremens is characterized with "Galgenhumor", namely the feeling of mixture of anxiety and euphoria. Fifty-nine patients with alcohol withdrawal acute psychosis, who admitted in Serigaya Hospital, Kanagawa Prefectural Center of Psychiatry, during the period August 1991-January 1993, were investigated clinically in focusing on the emotional condition. The emotional condition has been grasped in following four categories. 1. Moderate to intense anxiety, at least once during the course of withdrawal psychotic state: The patient shows manifest anxiety and irritability on his expression or behaviour and he complains his feeling of fear. 2. Only slight anxiety: The patient affirms his inner anxiety and fear, if he is asked. He is sometimes slightly restless but spends the time in the single room without any trouble. 3. No anxiety throughout the course of acute psychotic state. 4. Euphoric mood in the foreground, included cases which have slight anxiety together sometimes. Of 59 cases 6 belonged to the moderate and intense anxiety-group, 23 cases to the slight anxiety-group, 24 cases to the no anxiety-group and 6 to the euphoric group. This result indicates that anxiety in the alcohol withdrawal acute psychosis may be fewer in frequency and lighter in intensity in comparison with the past reports. The material is divided into two groups, delirium tremens and acute hallucinosis, with regard to the confusion of thinking and behaviour.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234535 TI - [On holding power of patients under voluntary admission--in relation to significance of voluntary admission system]. PMID- 8234536 TI - [Current problems with attitudes and informed consent on ECT: a questionnaire survey of psychiatrists]. PMID- 8234537 TI - [Reliability and validity of the Japanese version of the coping inventory for stressful situations (CISS): a contribution to the cross-cultural studies of coping]. AB - OBJECTIVE: There has recently been a dramatic increase in the number of studies on coping behavior as an intervening variable between stress and health. Most of the available measures of coping are, however, psychometrically inadequate. We therefore decided to develop the Japanese version of the Coping Inventory for Stressful Situations (CISS) with special regards to its cross-cultural equivalence, reliability and validity. The CISS is a self-report measure of an individual's typical pattern of coping along three orthogonal dimensions of Task , Emotion-, and Avoidance-oriented coping; its reliability and validity have been well studied in North America, where it was originally developed. METHOD: We obtained the Japanese version of the CISS (J-CISS) by means of back-translation. In Study 1, we administered the J-CISS and the 12-item General Health Questionnaire (GHQ) to 33 Japanese university students twice with an interval of four weeks. In Study 2,550 Japanese high school students completed the J-CISS and the Maudsley Personality Inventory. RESULTS: The equivalence of the Japanese version with the original was ascertained by means of back-translation involving multiple, independent mental health professionals and by factor congruence between the two versions. A principal component factor analysis (Varimax rotation) of the Study 2 data allowed us to extract three factors, which were virtually identical to the original ones. The high corrected item-remainder correlations, internal consistency reliabilities and test-retest reliabilities all attested to the reliability of the J-CISS. In order to examine its content validity, we compared the J-CISS with two coping questionnaires that have been in use in Japan, and found that the J-CISS covered most of the coping styles in these two questionnaires. However, such coping styles as "giving up," "to lose is to win (a Japanese proverb)," "it is best to do nothing" were not included in the original CISS and hence in the J-CISS. The criterion validity of the J-CISS was examined both in terms of predictive validity and concurrent validity. In Study 1, those who scored below the cut-off of the GHQ at Time 1 but above the cut-off at Time 2 had significantly higher Emotion-oriented coping scores at Time 1 than those who remained below the cut-off of the GHQ at Times 1 and 2 (predictive validity). In Study 2, the J-CISS scales and the MPI scales showed theoretically predicted correlations (concurrent validity). The results of the factor analysis and the corrected item-remainder correlations were suggestive of high construct validity of the J-CISS. Moreover, the mean inter-item correlation was between .20 and .40 for each scale, indicating its homogeneity. Factor analysis of each scale revealed that each scale indeed contained only one factor. Correlations among the three scales of the J-CISS established that the three scales formed multi dimensional measures of coping. CONCLUSION: The results of our study indicate 1) that the obtained Japanese version of the CISS is to be regarded as final, 2) that coping styles can be measured in a consistent and reliable manner both in Japan and North America, and 3) that this cross-cultural equivalence as well as the other validity studies have further augmented the validity of the CISS itself. PMID- 8234538 TI - [The substituted judgment to medical treatment and the guardianship]. PMID- 8234539 TI - Changes in sleep patterns by intralaminar thalamic microinjection of fluoxetine in rats. AB - Sleep patterns were investigated in rats by analysis of EEG recorded for 6 hours following fluoxetine or saline (control) microinjection into the centromedial intralaminar thalamus (CIT) at either 9:30 AM or 1:30 PM, 4 or 8 hours after lights on, respectively. The treatment of fluoxetine at 9:30 AM did not affect any sleep patterns. In contrast, fluoxetine significantly suppressed total paradoxical sleep (PS) time and the number of PS episodes during the second and third hour after administration at 1:30 PM. In addition, microinjection of fluoxetine produced an apparent enhancement of the duration of slow wave sleep (SWS) episodes and a small non-significant increase in total SWS time. On the other hand, no obvious changes in the basal sleep-wakefulness pattern were found between the two saline-treated control groups. These results indicate that CIT microinjection of fluoxetine at different time points acts differentially in sleep patterns and suggest that CIT may be one of the action sites for peripherally administered fluoxetine to alter sleep patterns. PMID- 8234540 TI - Effect of adenosine blockade on plasma renin activity and catecholamines. AB - We explored the hypothesis that chronic blockage of adenosine (Ado) receptors might augment the renin response to sodium restriction. In addition, an ancillary topic has been undertaken to test the hypothesis that endogenous Ado would restrain plasma norepinephrine (NE) and epinephrine (E) release. Male Sprague Dawley rats (250 +/- 20 g) were fed with low sodium chow (Na+ 0.03%) and drank either distilled water or caffeine containing water (0.1%). Blood pressures were recorded in the presence of different concentrations of Ado. Plasma renin activity was measured at the beginning, 1 week and 6 weeks after the low sodium diet. An in situ autoperfused rat kidney model was used to examine whether renal function was influenced by chronic inhibition of Ado receptors. Furthermore, the effect of theophylline on cardiovascular actions of Ado in anaesthetized rats, and the effect of 1,3-dipropyl-8-(p-sulfophenyl)xanthine (DPSPX) on acute hydralazine induced NE and E release in conscious, unrestrained rats were also determined. Our data showed that the inhibitory effect of Ado on renin activity and blood pressure in salt restricted rats was attenuated by caffeine at the first week but not at six weeks after institution of the low sodium diet. Renal function in sodium restricted rats did not change after chronic caffeine consumption. Theophylline attenuated the dose-related inhibitory actions of Ado on blood pressure. Finally, DPSPX significantly increased plasma catecholamines, and these effects were much greater in rats treated with hydralazine. We conclude that the augmented renin response is not continued during chronic blockage of Ado receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234541 TI - Beneficial effects of the opiate antagonist naloxone on hemodynamics and ventricular function following coronary artery occlusion and reperfusion in the dog. AB - It has been shown that endogenous opioid peptides (EOP) subserve important roles in cardiovascular regulation and are involved in the pathophysiology of myocardial ischemia, contributing to the deleterious effects. The aim of this study was to evaluate the effects of the opiate antagonist naloxone on the hemodynamics and ventricular function following coronary artery occlusion and reperfusion in the dog, utilizing the technique of cardiac catheterization. During myocardial ischemia and reperfusion, it was found that in the control group, there was reduction in the aortic, left ventricular, right atrial, pulmonary arterial and wedge pressures, and in the left ventricular dP/dt. The reduction in the aortic, left ventricular and pulmonary arterial pressures, and left ventricular dP/dt were significantly attenuated by pretreatment with naloxone. The results indicate a regulatory role of EOP in the cardiovascular function and suggest a possible involvement of EOP in myocardial ischemia and reperfusion causing detrimental effects such as arrhythmias, bradycardia, hypotension and, as shown in this study, impaired hemodynamics and ventricular function. The beneficial effects of naloxone on circulatory dynamics may have clinical implications in the prevention and treatment of ischemic heart disease. PMID- 8234542 TI - Influence of vegetarianism on fibrinolytic activity. AB - Blood glucose, cholesterol and triglyceride levels were determined and the venous occlusion test (VOT) was administered to 15 vegetarians (mean age 46.2 years, SD 15.1) and 20 non-vegetarians (mean age 38.3 years, SD 8.3). The tissue plasminogen activator (tPA), plasminogen activator inhibitor (PAI) and euglobulin lysis time (ELT) were measured before and after VOT. The means of all the parameters were not significantly different between vegetarians and non vegetarians. TPA increased after VOT in both the vegetarians and the non vegetarians, but the changing amplitudes of tPA, PAI and ELT induced by VOT were not significantly different in either group. This study evidenced no significant difference in the changes of fibrinolytic activity between vegetarians and non vegetarians. PMID- 8234543 TI - Complete nucleotide sequence and identification of a putative promoter region for the expression in Escherichia coli of the cryIA(b) gene from Bacillus thuringiensis var. aizawai HD133. AB - The sequence of a cry gene from Bacillus thuringiensis var. aizawai HD133 was determined. This cry gene encodes a protein of 1155 amino acids, the molecular weight of which is 130622 Da. When the nucleotide sequence of this cry gene was compared with the nucleotide sequence from B.t. var. aizawai IPL7 and var. berliner 1715, only five nucleotide changes were found. Therefore, this cry gene should be grouped into the cryIA(b) gene type. A 72 nucleotide sequence upstream of the open reading frame of the cry gene was enough to conduct the transcription of the cry gene in E. coli. Using a promoter probe vector system, it was demonstrated that any DNA fragment containing this 72 nt sequence exhibited promoter activity in vivo. It is likely that a putative promoter sequence may be present within this 72 nucleotides region for the expression of this cry gene in E. coli. PMID- 8234544 TI - Poor insight in schizophrenia. PMID- 8234545 TI - Do psychiatric patients need greater protection than medical patients when they consent to treatment? AB - It is commonly assumed that psychiatric patients require greater protection than medical patients when they make health care decisions. A review of the literature reveals that there is meager evidence for this belief and that both groups have significant capacity problems. Many factors other than psychiatric illness have been shown to impede understanding of informed consent information in health care settings. These include powerful emotional states, lack of a high school education, the presence of a severe medical illness, and increasing age. Clinicians can help patients become more adequate decision makers by taking such steps as providing adequate disclosure and inquiring about patient understanding. Even then, setting a realistically lenient standard of capacity seems the most practical approach. If screening for capacity is important for a specific health care decision, all vulnerable groups should be targeted. PMID- 8234546 TI - Genetic and biologic risk factors for suicide in depressive disorders. AB - Over the last few years data have accumulated suggesting that there may be genetic and biologic risk factors for suicide. This paper reviews studies from clinical populations, twins, the Amish, as well as data from the Iowa-500 and Danish-American adoption studies which suggest that there may be a genetic factor predisposing to suicide. The paper also reviews neurochemical data from cerebrospinal fluid, urine and postmortem studies which suggest that diminished central serotonin and dopamine may be implicated in suicidal behavior. PMID- 8234547 TI - Hippocampal atrophy in early Alzheimer's disease: anatomic specificity and validation. AB - We evaluated three groups of elderly individuals who were carefully screened to rule out clinically significant diseases that could affect cognition. They were matched for age and education. The groups included normals (N = 18), Alzheimer's Disease (AD) patients (N = 15), and minimally impaired individuals with memory complaints and impairments but who did not fulfill criteria for AD (N = 17). Volumetric measurements of different regions of the temporal lobe on the coronal scan as well as ratings of the perihippocampal cerebrospinal fluid (CSF) accumulation (HCSF) on the negative angle axial MR were carried out. Volume reductions were found in AD relative to the normals for both medial and lateral temporal lobe volumes. Only hippocampal volume reductions were found in the minimal group. The minimally impaired individuals had equivalent hippocampal volume reductions and significantly larger parahippocampal and lateral temporal lobe gyri than the AD group. The axial HCSF was validated using the coronal volumes. The combination of coronal hippocampal and perihippocampal CSF was the best predictor of the axial HCSF rating. The parahippocampal volume did not add to the predictive ability of the hippocampal-perihippocampal CSF combination. Future work should validate these findings with longitudinal designs as well as assess the issue of normal aging of these structures and their relationship to cognitive function. PMID- 8234548 TI - The temporal framework and Lacan's concept of the unfixed psychoanalytic hour. AB - Lacan's controversial clinical innovation of the unfixed analytic hour is evaluated within the context of Lacan's own theoretical and metaclinical views and against the classic, developmentally oriented concept of the fixed temporal analytic framework. Two clinical illustrations featuring disturbed reactions to the temporal framework of the analytic hour elucidate Lacan's proposed master/slave analogy for the treatment relationship, bringing to the fore, ironically, the basic contradictoriness of Lacan's clinical recommendation. PMID- 8234549 TI - The social matrix of aggression. Enactments and representations of loving and hating in the first years of life. AB - In this paper, we build on a developing body of work that addresses the various contributions to the child's emerging definition of self in the first five years of life. Described is the role of aggression in the shaping of the child's predominant modes of viewing and experiencing the world. How aggression toward another mixes with desire for another in the formation of early capacities for object relatedness is a central question for any developmental theory of aggression, for aggression is first experienced, shaped, refined, and remodeled in the context of loving relations. Studies from related disciplines provide observational data about how aggression is modified in the first five to six years of life and how the capacity to interpret the intentions of others is crucial to those modifications. Understanding the intentions of others relates also to the capacity to attribute affects, beliefs, and other mental states to others, a capacity referred to elsewhere as a theory of mind. The ability to reflect upon thoughts and feelings in oneself and others provides children with a fuller range of responses to their own and others' perceived aggression. Finally, external experiences of violence, abuse, and deprivation influence the child's experience of his own and others' aggression and prevent the normal modulation of aggression in the social matrix. PMID- 8234550 TI - Problems in female development. Comments on the analysis of an early latency-age girl. AB - A 6 1/2-year-old girl's clinical material relevant to the emergence of clitoral and vaginal masturbation and masturbation fantasies during analytic sessions is presented. The intense genital excitation, which she called "jeebies," often overwhelmed her ego functioning. She assumed the jeebies and the fantasies that precipitated and accompanied them would cause bad things to happen to her. External events and the nature of her object relations reinforced and confirmed her belief that being only female was unacceptable. Her penis envy, a wish to be a boy-girl, had multiple determinants. A detailed description of her evaluation and subsequent treatment reveals how her unconscious conflicts were resolved in analysis through the evolution and resolution of her transference neurosis, validating the psychoanalytic approach. PMID- 8234551 TI - The impact of divorce on a preadolescent girl. AB - This paper presents clinical material from the psychoanalysis of a 12-year-old girl whose parents had divorced several years earlier. Material drawn from her analysis permits us to examine the question: given the intrapsychic demands of the prepubertal period, how was the experience of divorce registered, what aspects of intrapsychic functioning became distorted, and what, if any, remained uncontaminated? The impact of the parental divorce lay in an intensification and entrenchment of characteristic latency defenses which, as puberty approached, contributed to heightened internal conflict and an insistent turn to reality as both explanation for and solution of intrapsychic conflict. PMID- 8234552 TI - Reality, fantasy, and adolescence. AB - Psychoanalytic views of reality and fantasy are contingent upon a theory of ego/mind: A mental-apparatus-objective approach is distinguishable from an experiential-subjective approach. These two views are highly correlated with Being and Doing as underlying modalities of processing experience. Adolescent development necessitates the integration and amalgamation of these two modalities, which are reflected in relating to reality as objective and separate from, or as merged and fused with, the ego, with corresponding fantasy contents and ego states. As social reality exists as an independent variable, it poses real problems for such crucial aspects of adolescence as identity formation, psychosocial moratorium, and the generational gap. PMID- 8234553 TI - Play in adulthood. A developmental consideration. AB - This paper is about normal development, addressing the basic characteristics and evolution of play throughout life, with particular emphasis on the nature of play in adulthood. Although the psychoanalytic literature on play in childhood is extensive, undoubtedly because of its relevance to child analysis, very little has been written on the subject of adult play or on the relationship between adult play and its childhood antecedents. PMID- 8234554 TI - Preventive intervention with the children of depressed parents. A case study. AB - Children of parents with depression are at significant risk for impairment, but little is known about preventive intervention strategies prior to the onset of illness in the children. This paper discusses a preventive approach to assisting families in helping children cope with parental affective disorder. A case study is presented which demonstrates the compatibility of this family-based intervention with psychoanalytically oriented treatment for the parent. Key aspects of this work include the enhancement of self-understanding and resiliency in children. PMID- 8234555 TI - The reaction of an early latency boy to the sudden death of his baby brother. AB - Surprisingly little has been written on the impact of the death of a baby on an older sibling. This paper describes how a narcissistically vulnerable latency boy grappled, in the course of his psychotherapy, with the painful loss of his baby brother. Emotional and cognitive aspects of his early confrontation with the reality of death are considered. The patient's six theories about the cot death are described. Attention is drawn to the value of the sibling relationship and to the narcissistic needs which this relationship fulfilled. The termination phase is discussed in the light of the dynamics of sudden loss. The safe therapeutic context provided the space in which this young, rather muddled latency boy could grieve and mourn this traumatic loss. PMID- 8234556 TI - Telling and enacting stories in psychoanalysis and psychotherapy. Implications for teaching psychotherapy. AB - This paper is about teaching psychoanalytic psychotherapy. The approach to teaching is based on a view of the patient's plight or psychopathology, and on a way of listening to psychotherapeutic process, that focus on the stories a patient tells and enacts. The author relates how various experiences in his life led him to this narrative paradigm. PMID- 8234557 TI - How people get better. PMID- 8234558 TI - Portraits of survival. A twenty-year follow-up of the children of Buffalo Creek. AB - This study is a follow-up of the children of Buffalo Creek "hollow" who survived the dam collapse and flood of 1972. It was conceived as a complement to the 1988 NIMH-funded follow-up investigation of the children of Buffalo Creek conducted by the University of Cincinnati Traumatic Stress Study Center. That 1988 study utilized standardized methodology to assess levels of psychopathology present among those who were children at the time of the 1972 flood. Among the original child subjects, results demonstrated that the rates of both posttraumatic stress disorder (PTSD) and levels of other psychopathology had sharply declined as the children moved to adulthood. Several questions remained unanswered: the long-term meaning which the flood had in the survivors' lives and its impact on their adaptation as they progressed through the subsequent stages of emotional development. For these reasons, we, as psychoanalytic investigators who had been members of the original clinical assessment team, planned an interview of child and adolescent survivors as a follow-up to our interviews in 1974 with a particular focus on meaning and adaptation. PMID- 8234559 TI - Reflections on a year's psychotherapy with a psychotic man. AB - A year's psychotherapy with a psychotic man during the author's residency training is described in detail. Fundamental questions regarding the patient's potential for therapeutic change are raised. These questions concerning this specific patient are then generalized to broader research questions regarding the efficacy of treatment for the severely and persistently mentally ill. PMID- 8234560 TI - Family romance fantasy resolution in George Eliot's Daniel Deronda. PMID- 8234561 TI - Insights into the struggle of creativity. A rereading of Anna Freud's "Beating fantasies and daydreams". AB - The wish to create, the dilemmas regarding the value, source, and ownership of one's potential and ideas, and the threats that these involve, are explored as they emerge in a dialogue between Anna Freud and her father, embedded in Anna Freud's first paper "Beating Fantasies and Daydreams" (1922). Insight into these aspects of the struggle of creativity contributes to the understanding of their relationship to writing, masochism, sublimation, and other associated psychological processes. Through the analysis of the paper the identity of the patient described is revealed to be Anna Freud herself. This sheds light on her personal development and the way in which it influenced developments in psychoanalytic thinking. PMID- 8234562 TI - The roles of mental representations and mental processes in therapeutic action. AB - In this paper we describe two models of the psychoanalytic treatment of mental disturbance. The first describes the mechanism by which the patient is helped to recover threatening ideas and feelings which have been repudiated or distorted as a result of conflict and defense. The second points to the therapeutic effects of engaging previously inhibited mental processes within the psychoanalytic encounter. The two forms of therapeutic action imply two distinct means available to the individual to deal with psychological conflict. They highlight different aspects of the psychoanalytic process and technique in child and adult psychoanalysis. PMID- 8234563 TI - The theoretical contributions of Hans W. Loewald. AB - An integration and dialectical reading of Hans Loewald's developmental theory is offered, demonstrating his ability to bridge theoretical dualisms. He maintains yet revises Freud's structural model and dual instinct theory in a way which embraces developmental and relational perspectives. PMID- 8234564 TI - Mental disorder and the strategies of prevention. PMID- 8234565 TI - Noradrenergic modulation of cognitive function: clinical implications of anatomical, electrophysiological and behavioural studies in animal models. PMID- 8234566 TI - A modern melancholia: Van Gogh's portrait of Dr Gachet. PMID- 8234567 TI - Reduction of cortical volume in schizophrenia on magnetic resonance imaging. AB - The MRI scans of 48 schizophrenic patients, fulfilling RDC criteria, were compared to those of 34 healthy controls matched for age, ethnicity and parental social class. The volume of the frontal and anterior parietal lobes was significantly reduced in the schizophrenic group as a result of a selective decrease in cortical volume, with a corresponding increase in the volume of sulcal fluid. Reduction in the volume of the temporal grey matter was more marked on the right, but was not in excess of the loss of volume observed in other areas of the cortex. MRI abnormalities correlated poorly with clinical parameters, although both unemployment and poor pre-morbid adjustment predicted reduced cerebral volume and increased sulcal volume. These results question whether the medial temporal lobes are the only site of structural pathology in schizophrenia. PMID- 8234568 TI - The Edinburgh cohort of HIV-positive drug users: the relationship between auditory P3 latency, cognitive function and self-rated mood. AB - One hundred and six HIV-positive drug users were tested with a two-tone auditory evoked potential (AEP) task and a small battery of neuropsychological tests, to examine the relationship between the latency of the P300 component (P3) of the AEP, intellectual function, mood and drug use. Principal components analysis revealed a significant correlation between P3 latency and the first principal component (r = -0.43, P < 0.001). Varimax rotation generated three orthogonal components which we interpreted as intellectual performance, memory, and mood. Intellectual performance and self-reported mood were individually correlated with P3 latency, but memory was not (r = -0.36, P < 0.001; r = 0.23, P < 0.05; and r = -0.18, NS, respectively). Subjects with symptomatic HIV disease had a higher correlation between P3 latency and intellectual performance than subjects with asymptomatic HIV disease and, among patients with symptomatic HIV disease, poorer memory was associated with a lower CD4 count. Opiate or benzodiazepine consumption did not correlate with poor intellectual performance, memory, or self rated mood in our sample. These results indicate that there is a relationship between AEP latency and neuropsychological measures of intellectual function, and that it is influenced by subjective mood. Surprisingly, declared current drug use has no discriminable effect on these relationships. PMID- 8234569 TI - Neuropathological basis for drawing disability (constructional apraxia) in Alzheimer's disease. AB - The performance on four drawing tasks was studied in a sample of patients with verified Alzheimer's disease in order to examine the relationship of 'constructional apraxia' to neuropathological changes in the parietal lobe and in other brain areas. Twenty-three patients were able to attempt to copy pentagons, a spiral and a three-dimensional drawing of a house, 22 patients were able to draw a clock-face spontaneously. The results were rank-ordered by two independent raters. The values obtained in the different drawing tasks were correlated significantly with each other, with global estimates of cognitive performance (CAMCOG, Mini-Mental State), with a shorter duration of illness, higher brain weight (in the subsample of female patients), higher counts of large neurons in the parahippocampal gyrus and hippocampus, but not in the parietal lobe. This suggests that there is no specific relationship between 'constructional apraxia' and neuropathological changes in the parietal lobes of patients with advanced Alzheimer's disease, but that there is a correlation between widespread brain changes and several neuropsychological deficits, one of them being drawing disability. PMID- 8234570 TI - Clinically diagnosed presenile dementia of the Alzheimer type in the Northern Health Region: ascertainment, prevalence, incidence and survival. AB - Potential cases of presenile dementia of the Alzheimer type (PDAT) in the Northern Health Region (estimated population aged 45-64, 655,800) were ascertained for the years 1979-86 from in-patient ICD-9 codes and other sources. An algorithm was applied to the casenote information to distinguish between Alzheimer-type and other forms of dementia. A search of the NHS central register was made to establish date and place of death. Estimates were made for patients with missing case records. The point prevalence rate for PDAT was estimated as 34.6 per 100,000 with an annual incidence of 7.2 per 100,000 in the 45-64 age range. These rates are compared with those reported in other studies. Five-year survival following diagnosis for incident cases of PDAT was 64% with a longevity quotient (LQ), the percentage of expected time actually survived, of 69%. There was no evidence of a more malignant course in PDAT when compared with survival in older patients with dementia of the Alzheimer type (DAT) in other studies. Sixty six per cent of deaths occurred in hospital, 19% at home and 15% in residential homes. PMID- 8234571 TI - War-induced prolonged stress and metabolic control in type 2 diabetic patients. AB - Sound evidence that psychological stress may significantly worsen metabolic control in diabetic patients in lacking. The stressful effects of war on glycaemia control have not been assessed as yet. A randomly selected sample of displaced Type 2 diabetic persons (N = 44) was compared to a matched group of diabetic persons who had not been forced to leave their homes. The self-reported stress, depression level, fasting and post-prandial blood glucose, glycosylated haemoglobin and serum lipids were compared. The two groups were found to be significantly different in scores for self-reported stress and depression levels (P < 0.001). The proportion of subjects belonging to the category of clinically significant depression was larger in the group of displaced persons (P < 0.001), as well as the proportion of extreme scores for self-reported stress (P < 0.001). No significant differences between the groups were found for variables measuring metabolic control (P > 0.05), except for the serum triglycerides which were found to be significantly higher in the group of displaced persons (P < 0.01). This is assumed to be the result of differences in nutrition. The results do not demonstrate an association between prolonged stress and glycaemic control in Type 2 diabetic patients. The variability of data measuring stress and depression level in two extreme groups with respect to HbA1c values suggests that individual factors which determine the response to stressors need to be explored. PMID- 8234572 TI - Psychosocial stress and breast cancer: the inter-relationship between stress events, coping strategies and personality. AB - This quasi-prospective study of 2163 women attending a breast-screening clinic for a routine medical check-up indicates that certain types of coping strategies and personality dispositions predispose some women to an increased risk of developing breast cancer following the occurrence of a major life-event such as bereavement or other loss-related event. Regular exposure to stress situations appears to reduce the risk of a malignancy; experiencing a single, major life event was found to be potentially much more damaging, particularly if the individual was unable to externalize her emotions and obtain appropriate help and counselling. PMID- 8234573 TI - Setting the scene for eating disorders: childhood care, classification and course of illness. AB - The aim of this study was to determine whether the childhood experiences of patients with anorexia nervosa and bulimia nervosa differ and affect the course of the illness. A semistructured interview developed by Harris et al. (1986) was used to assess the childhood family environment of 64 patients with restricting anorexia nervosa (RAN), 23 patients with bulimic anorexia nervosa (BAN), 37 bulimic patients with a history of anorexia nervosa (BN/HistAN) and 79 patients with normal weight bulimia nervosa (BN). There were no significant differences between groups in terms of parental mental disorder, low parental control or childhood sexual abuse. BN patients had had significantly more family arrangements and had experienced more parental indifference, excessive parental control, physical abuse, and violence against other family members than RAN patients with the BAN and BN/HistAN group being intermediate. There was a trend for BN-patients to have had more intra-familial discord than the other groups. Different aspects of adversity tended to cluster in the same patients and 65% of the bulimic group had experienced two or more types of childhood adversity. These results suggest that childhood experiences contribute to the form of eating disorder which later develops. PMID- 8234574 TI - Anger suppression: its relationship to beta-adrenergic receptor sensitivity and stress-induced changes in blood pressure. AB - While studies from diverse fields of research suggest a relationship between problems expressing anger and cardiovascular illness, few studies have provided a potential pathophysiological link of such a relationship. Forty-five males were classified according to one of three anger expression categories: those who did not suppress their anger (N = 13), those who partially suppressed their anger (N = 19), and those who definitely suppressed their anger (N = 13). For each, we determined lymphocyte beta-adrenergic receptor function and blood pressure responsiveness to a standardized mathematics stressor. Those subjects who routinely suppressed their expression of anger had increased beta-adrenergic receptor sensitivity (P = 0.01) (isoproterenol-stimulated cyclic AMP production) and a greater systolic blood pressure response to the stressor (P = 0.001). Anger suppression was unrelated to the subject's age, weight, or socioeconomic status. These findings may be germane to prior clinical and epidemiologic observations relating anger expression and cardiovascular illness. PMID- 8234575 TI - Childhood adversities and adult depression: basic patterns of association in a US national survey. AB - The long-term relationships between various forms of childhood adversity and adult episodes of major depression are explored in a representative household survey of the United States adult (age 25+) population. Seven of the eight childhood adversities considered are significantly associated with recent (12 month) episodes of depression. These effects are largely indirect consequences of some childhood adversities leading to a life history of depression and prior depression leading to new episodes. Only three of the eight childhood adversities directly affect recent onset or recurrence. The paper closes with a discussion of implications for future research on the long-term effects of childhood adversities. PMID- 8234576 TI - An epidemiological study of mental disorders in a 10-year cohort of young adults in Israel. AB - This is an overview of a two-phase epidemiological study of mental disorders among young adults in a ten-year birth cohort (1949-58) conducted in Israel. A sample of 4914 Israel-born offspring of Jewish immigrants was obtained by full probability sampling procedures and screened for caseness using psychometric symptom scales from the Psychiatric Epidemiology Research Interview (PERI). Those screened positive and almost a fifth of the negatives (N = 2741) were interviewed by psychiatrists using the Schedule for Affective Disorders and Schizophrenia, Israel version (SADS-I), in order to determine prevalence rates of specific disorders as defined by the Research Diagnostic Criteria (RDC). The completion rates for each interview phase were 94.5% and 90.7% respectively. Six-month prevalence rates are presented by gender, ethnic origin, and education. Approximately one-fifth of the birth cohort met current RDC criteria for a disorder at the definite level, excluding the RDC category of 'other psychiatric disorder'. Generalized anxiety disorder and major depressive disorder were the most commonly found types. The striking findings centre on alcoholism and drug use disorder which were exceedingly rare, and the unusually similar rates of major depression for males and females. The results from this study are discussed in comparison with those obtained from other epidemiological studies. PMID- 8234577 TI - The epidemiology of social phobia: findings from the Duke Epidemiological Catchment Area Study. AB - Social phobia was studied in a North Carolina community, using DSM-III criteria. Two kinds of comparison were made: social phobia v. non-social phobia, and comorbid social phobia v. non-comorbid social phobia. Six-month and lifetime prevalence rates were 2.7 and 3.8% respectively. Social phobia had an early onset, lasted a long time and rarely recovered. Predictors of good outcome recovery in a logistic regression analysis were onset of phobia after age 11, absence of psychiatric comorbidity and greater education. The disorder was often missed in medical consultation. Increased rates of psychiatric comorbidity existed, especially for other anxiety disorders and for schizophrenia/schizophreniform disorder. There was increased risk of neurological disorder. Social phobia was also associated with an increased rate of suicide attempts, antisocial behaviour and impaired school performance during adolescence, impaired medical health, increased health-seeking behaviour, poor employment performance, reduced social interaction and impaired social support. Comorbidity accounted for some, but not all observed differences. PMID- 8234578 TI - The prevalence of depressive disorders and the distribution of depressive symptoms in later life: a survey using Draft ICD-10 and DSM-III-R. AB - The point prevalence of depressive disorders was estimated in a sample of persons aged 70 years and over, which included both those living in the community and those in institutional settings. Lay interviewers administered the Canberra Interview for the Elderly to the subjects and their informants. The point prevalence of depressive episodes as defined by the Draft ICD-10 diagnostic criteria was 3.3%. The rate for DSM-III-R major depressive disorder was 1.0%. The latter prevalence rate is similar to those reported elsewhere for the elderly. Evidence is accumulating that older persons may indeed have low rates for depressive disorders at the formal case level. Possible reasons for this finding are offered. A scale for depressive symptoms, based exclusively on those specified in Draft ICD-10 and DSM-III-R, showed that the elderly do experience many depressive symptoms. Contrary to expectation, these did not increase with age. The number of depressive symptoms was correlated with neuroticism, poor physical health, disability and a history of previous depression. Attention now needs to be directed to the clinical significance of depressive symptoms below the case level in elderly persons. PMID- 8234579 TI - Unemployment, underemployment and depressive affect among Southeast Asian refugees. AB - This study of 1348 adult Southeast Asian refugees resettling in Vancouver, British Columbia and a comparison sample of 319 permanent residents of the city demonstrates a reciprocal relationship between unemployment and depressive affect. Increased risk of depression accompanied job loss and depression made it more difficult to stay employed. Although the association between unemployment and depression was common to both refugee and host society samples, links between these phenomena proved different in the two groups. In contrast to the refugees for whom income loss was the over-riding stress resulting from job loss, loss of esteem and loss of social contact also proved to be salient stressors for resident Canadians. Although a threat to the mental health of resident Canadians, underemployment--working at a level which considering one's education and previous occupation, is lower than might be expected--did not jeopardize the mental health of refugees. PMID- 8234580 TI - Symptom dimensions and cognitive and social functioning in recent-onset schizophrenia. AB - The relationships among symptoms, cognitive functioning and social functioning were investigated in 60 patients with recent-onset schizophrenia. Positive symptoms were unrelated to cognitive measures. Disorganization and depressive symptoms were correlated significantly with Card Sorting performance. Furthermore, only negative symptoms were correlated significantly with social functioning. These results replicate earlier studies with chronic and mixed samples, and support the validity of disorganization as a separate symptom dimension. In contrast to most previous studies, no significant correlations were found between negative symptoms and cognitive measures. However, some evidence was found for a non-linear association between negative symptoms had several cognitive measures. The variation explained by a curvilinear model was not high, but for some cognitive measures this model was clearly superior to a linear model. If replicated, this finding supports the position that cross-sectionally measured negative symptoms cannot be viewed as a unitary concept. PMID- 8234581 TI - Association between major depressive disorder and physical illness. AB - The association between major depressive disorder (MDD) and self-reported histories of specific physical illnesses was investigated in 320 controls and 1968 first-degree relatives and 254 spouses of probands in the NIMH Collaborative Depression study. The Schedule for Affective Disorders and Schizophrenia-Lifetime Version was used to assign Research Diagnostic Criteria (RDC) diagnoses and a structured self-report instrument was used to assess lifetime medical history. Lifetime MDD was diagnosed in 914 subjects, 402 of whom had been hospitalized or received somatic treatment ('treated' MDD). Strong associations were observed between MDD (either treated or untreated) and both frequent/severe headaches and migraine headaches. There was a marked gender effect such that the relative odds for a woman with treated MDD to report migraine were over 5:1. Other associations were found between MDD and skin infections, respiratory illness, ulcer, hypotension, and diabetes. This is the largest non-patient sample using standardized assessment of mental disorders by direct interview in which associations between specific physical illnesses and MDD have been demonstrated. Implications for clinical practice and neurobiological research in depression are discussed. PMID- 8234582 TI - Perceived health and high consumers of care: a study of mental health problems in a Swedish primary health care district. AB - Many patients with mental health problems are treated in primary health care services. They are often multi-users of care. In the present investigation we have studied patients visiting primary health care clinics who have been assessed for mental health problems according to the Hopkins Symptom Check List (HSCL-25). Two quality-of-life instruments (the Nottingham Health Profile and the Mood Adjective Check List) were applied to describe further the perceived health of the sample. A random sample of 93 patients was chosen out of 388 patients visiting a primary health care clinic in a Swedish health care district during four weeks in January and February 1990, and these patients were followed prospectively during one year. High consumers of health care during the follow-up period scored a poor perceived health. 'Feeling worthless', predicted a high number of days of sickness absenteeism. Hidden cases, not detected by the GPs, consumed more resources than those identified by the GPs. The importance of training and educating general practitioners in the early detection of patients with mental health problems is discussed. PMID- 8234583 TI - Expressed emotion and schizophrenia: the ontogeny of EE during an 18-month follow up. AB - Key relatives of 30 first or early admission psychotic patients (mainly with a diagnosis of schizophrenia) were interviewed (for a second time) 18 months after participating in an initial assessment of expressed emotion (EE). The patients were followed up throughout the interim period, during which time 17 experienced a psychotic relapse. Although there continued to be a negligible association between initial EE rating (established during the index admission) and relapse, a significant association between EE rating at 18 months and psychiatric status of the patient during the follow-up period emerged. Results are discussed in the context of recent interactional theories of the relationship between household EE and psychiatric morbidity. PMID- 8234584 TI - Inter-rater and temporal reliability of the Standardized Assessment of Personality and the influence of informant characteristics. AB - The Standardized Assessment of Personality (SAP) is a short, semi-structured interview designed to be used with an informant of the patient. The informants of 52 patients were interviewed separately by two raters for an inter-rater reliability study. The overall level of agreement was excellent, kappa = 0.76, with a range between 0.60 and 0.82 for the individual categories of personality disorder. For a temporal reliability study, the relatives of 77 patients were interviewed. The overall level of agreement was good, kappa = 0.65, with a range between 0.54 and 0.79 for individual categories. Characteristics of informants whose reports were rated with greater levels of reliability were female gender and greater length of acquaintance with the patient. PMID- 8234585 TI - Frontal lobe impairment in schizophrenia: relationship to intellectual functioning. AB - Schizophrenic subjects (N = 48) and individually matched healthy controls were administered the Verbal Scale of the Wechsler Adult Intelligence Scale (VIQ) and a test of verbal fluency. The verbal fluency and VIQ scores of the schizophrenic subjects were significantly lower than the scores of the control subjects. An additional sample of healthy subjects (N = 144) was used to generate a regression equation for the prediction of verbal fluency scores from Verbal IQ and age. The verbal fluency scores obtained by the schizophrenic subjects were significantly lower than the scores predicted from the regression equation, whereas a significant difference was not obtained in the matched controls. These results provide further evidence of frontal lobe dysfunction in schizophrenia. PMID- 8234586 TI - Psychiatry in the 1890s. PMID- 8234587 TI - F Scale elevation and PTSD MMPI profiles. AB - MMPI profiles for 87 PTSD veteran inpatients were classified and studied according to MMPI F Scale elevation. Mean MMPI profiles and frequency of high two point code types were studied for different levels of F Scale elevation. Similar mean profile configurations were found for subgroups with F > or = 70 with Scales 2 and 8 appearing as the two highest clinical scales. For F < 70 the configuration was different in that Scale 8 was not one of the two highest scales. The 2-8/8-2 high two-point code was the modal high two-point code for the total sample but the relative frequency of high two-point codes did vary somewhat within and across levels of F Scale elevation. PMID- 8234588 TI - Work stress, role conflict, social support, and psychological burnout among teachers. AB - This study examined a research model developed to understand psychological burnout among school-based educators. Data were collected from 833 school-based educators using questionnaires completed anonymously. Four groups of predictor variables identified in previous research were considered: individual demographic and situational variables, work stressors, role conflict, and social support. Some support for the model was found. Work stressors were strong predictors of psychological burnout. Individual demographic characteristics, role conflict, and social support had little effect on psychological burnout. PMID- 8234589 TI - Borderline personality disorder and suicidal behavior. PMID- 8234590 TI - Psychological consequences of genital herpes, an exploratory study with a gonorrhea control-group. AB - Measurements of some psychosocial variables were obtained from 27 patients with a genital herpes infection and compared with those from 12 patients with a gonorrhea infection. The measurements referred to the period before and during the infection. Evidence was found that during the infection patients were more anxious, more sexually inhibited, more bitter towards their partners, and had more psychological complaints than before the infection. Generally there was no difference between the two groups of patients, with one exception: patients with genital herpes judged themselves as having fewer psychological complaints prior to the disease than did patients with gonorrhea. PMID- 8234591 TI - Use of the MMPI to distinguish genuine from factitious multiple personality disorder. AB - MMPI special scales were repeatedly administered to several personality states of a woman alleging to have Multiple Personality Disorder. Her inability to produce a consistent response set on retesting helped to unmask a case of factitious multiple personality. PMID- 8234592 TI - Perceived crime, fear of crime, and walking alone at night. AB - This study examined longitudinal patterns in public opinion about crime, fear of crime, and walking alone at night. Data sets from the Winnipeg Area Study of 4050 randomly selected adults who were interviewed in their homes were used. The data reflect high concern by respondents for increasing crime rates. Analysis suggests that perceived crime in the city and in Canada were not associated with fear of crime and walking alone at night; perceived crime in respondents' neighborhoods showed a weak association. Implications for measures of perceived crime are discussed. PMID- 8234593 TI - Countertransference: a typological expansion. AB - Countertransference was viewed from the perspective of a 2 x 2 x n factorial typology (source x degree of consciousness x depth). This typology incorporated the conscious and unconscious influences of nonpsychological variables and their theoretical schema upon the analyst's therapeutic recommendations. PMID- 8234594 TI - Correlations among scales of the Beck Depression Inventory and the profile of mood states. AB - The revised Beck Depression Inventory and the Profile of Mood States were administered to 104 men. Scores on the Beck Depression Inventory correlated significantly with all six Profile scales, including .75 with Depression Dejection and -.53 with Vigor-Activity. PMID- 8234595 TI - Factors related to susceptibility and recruitment by cults. AB - Unprecedented escalation of secular and religious cults has necessitated further inquiry into more precise conditions under which individuals develop vulnerability and become converted by these groups. The present discussion focuses on a number of factors which seem to influence individuals' susceptibility and recruitment by cults. These variables include (a) generalized ego-weakness and emotional vulnerability, (b) propensities toward dissociative states, (c) tenuous, deteriorated, or nonexistent family relations and support systems, (d) inadequate means of dealing with exigencies of survival, (e) history of severe child abuse or neglect, (f) exposure to idiosyncratic or eccentric family patterns, (g) proclivities toward or abuse of controlled substances, (h) unmanageable and debilitating situational stress and crises, and (i) intolerable socioeconomic conditions. Also presented are methods utilized by cults, e.g., intimidation, coercion, and indoctrination, for systematically recruiting, initiating, and influencing inductees. More careful attention to these factors might help health care providers, educators, clergy, and concerned family and friends determine more precisely individuals at greater risk for recruitment into cults. PMID- 8234596 TI - Suicide and social status among Native Americans. PMID- 8234597 TI - Memory of Northern Irish Catholics and Protestants for violent incidents and their explanations for the 1981 hunger strike. AB - The aim of this study was to examine the memory of Northern Irish Catholics (n = 20) and Protestants (n = 21) for violent events which had occurred over the previous 11 years and their explanations for those events. It was predicted that Catholics would recall more events involving Catholic deaths than Protestants and that Protestants would recall more events involving Protestant deaths than Catholics. Although Catholics were as likely as Protestants to recall incidents which resulted in Protestant deaths, Protestants were less likely than Catholics to recall incidents involving Catholic deaths. Also, there were divergent explanations for the 1981 hunger strike with most Protestants attributing responsibility to factors internal to the hunger strikers and most Catholics attributing responsibility to factors external to the hunger strikers. PMID- 8234598 TI - Cognitive abilities and lipomyelomeningocele. AB - Ten children with lipomyelomeningocele were evaluated with the WISC--R, the Wide Range Achievement Test--Revised, the Developmental Test of Visual-motor Integration, and the Child Behavior Checklist. These children were consecutive referrals to a birth defects clinic. Unlike their meningomyelocele counterparts, as a group these children appear to be average in their intellectual, academic, and behavioral characteristics. However, they exhibited low average perceptual motor skills, a feature more commonly seen in meningomyelocele. PMID- 8234599 TI - Sexual fears are increasing. AB - This study compared scores from three samples of college students (N = 810) on the Sexual Aversion Scale, a measure of sexual anxiety based on DSM-III-R criteria for diagnosing sexual aversion disorder. Measurements were taken in 1988, 1991, and 1992. There was a significant increase in sexual anxiety during this period, most of it attributable to fear of acquiring AIDS. Although the women reported more sexual anxiety than the men over-all, there was no difference on fear of AIDS. These findings could foreshadow an increase in sexual dysfunctions and desire disorders. PMID- 8234600 TI - Relationship of loneliness and its affective elements to self-esteem. AB - This study was designed to investigate the relationship between loneliness and self-esteem for adolescents (n = 57; age range 16 to 18 years). As hypothesized, scores on the UCLA Scale for Loneliness correlated negatively with adolescents' scores on Coopersmith's self-esteem measure. And, scores on two affective dimensions of loneliness (Isolation and Dejection) correlated negatively with scores on self-esteem. PMID- 8234601 TI - AIDS and college students: a survey of knowledge, attitudes and beliefs. AB - A survey of beliefs about, knowledge of, and attitudes about AIDS was administered to 345 college students. While knowledge was generally high, reported behavior was not always consistent with that knowledge. PMID- 8234602 TI - Self-induced water intoxication in mental patients. PMID- 8234603 TI - Accurate knowledge about suicide and personality. PMID- 8234604 TI - Perspective reports of corporal punishment by pupils in Lesotho schools. AB - This study surveyed reports of practices of corporal punishment at secondary schools in Lesotho by 60 randomly selected pupils. There were 34 males and 26 females, whose mean age was 21 years, with a range between 14 and 29 years. Responses to a questionnaire confirmed that punishment was associated with pupils' reports of academic impairment, psychological damage, and physical injury. PMID- 8234605 TI - Dissociative experiences of Vietnam veterans with chronic posttraumatic stress disorder. AB - Interest in dissociation has been renewed, and its relationship to Post-traumatic Stress Disorder is especially intriguing. In this study 57 consecutively admitted chronic, combat-related Posttraumatic Stress Disorder sufferers were grouped by scores on a dissociative scale (Dissociative Experiences Scale). The three groups (high, medium, and low) were compared on personality measures (MMPI basic scales and subscales, and Millon's MCMI), Posttraumatic Stress Disorder measures, and a psychophysiological index of heart rate under baseline trauma conditions. The results showed that the survivors with more dissociative experiences show distinctive and higher symptom levels--excessive fearfulness, symptoms of strange experiences, and high tonic psychophysiological states--as well as greater severity of ratings of Posttraumatic Stress Disorder (on the Mississippi Scale). The discussion addressed the possible role of dissociation in Posttraumatic Stress Disorder. PMID- 8234606 TI - A psychological "sense-of-control" profile of patients with anorexia nervosa and bulimia nervosa. AB - To investigate control and self-control issues for patients with eating disorders, 10 individuals diagnosed with anorexia nervosa or bulimia nervosa were compared to two sex-matched groups of 9 and 50 adults on the Shapiro Control Inventory. Analysis of variance and subsequent planned comparisons showed significant differences indicative of pathology between the eating-disordered group and one or both comparison groups on the general domain sense of control scale and the positive sense of control scale. Three of the four mode scales and the domain-specific sense of control scale were also in the expected direction. Several clinically relevant individual items also supported this trend. The discussion notes the clinical importance of refining how control is measured for eating-disordered populations. PMID- 8234607 TI - Practical applications from chaos theory to the psychotherapeutic process, a basic consideration of dynamics. AB - This essay contains speculations about the import of some ideas from chaos theory for psychotherapeutic process. Brief consideration is given to areas of psychology in which such ideas have been applied, as well as some ideas on psychic energy. Then, using a psychodynamic perspective as a base, three aspects of psychotherapy are addressed, the container, the process, and the clinician as the agent of change. A central idea is that the emotional experience of chaos is a necessary developmental step. It is important for the therapist to recognize whether a client is avoiding this experience or is overwhelmed by this experience. PMID- 8234608 TI - Factors affecting the progression towards burnout: a comparison of professional and volunteer counsellors. AB - This study compared professional and volunteer counsellors on a number of key variables in the progression to burnout, namely, stressors, strain, defensive coping, social support, Type A personality, and breakdown. Job stressors unique to each group were identified but there was overwhelming evidence for professionals and volunteers belonging to a homogeneous counselling population as evidenced by similarity across major constructs. Path analysis supported theoretical formulations on burnout by Maslach and Cherniss, and gave support to Lin, Dean, and Ensel's argument that social support may act as a buffer to burnout. PMID- 8234609 TI - Self-report versions of the Yale-Brown Obsessive-Compulsive Scale: an assessment of a sample of normals. AB - The Yale-Brown Obsessive-Compulsive Scale was administered to 180 undergraduates and 50 medical clinic patients to obtain normative data for this widely used instrument. These data should prove useful to clinicians who assess obsessive compulsive symptomatology. PMID- 8234610 TI - Relationship of self-esteem, manifest anxiety, and obsessive-compulsiveness to personal habits. AB - 75 women and 64 men responded to the Coopersmith Self-esteem Inventory, the Manifest Anxiety Scale, and the Maudsley Obsessional-compulsive Inventory in addition to responding to a questionnaire on personal habits. The results indicated that more frequent hair-pullers and nervous twitchers scored lower on self-esteem and higher on anxiety. People who giggled and those who bit their fingernails more often scored higher on obsessive-compulsiveness. Self-reported gigglers were higher on manifest anxiety. If the criterion of self-assessed seriousness of the behavior problem was used, people who bit their nails, picked their noses, pulled their hair, chewed on objects, giggled, ground their teeth, twitched nervously, and picked at scabs scored lower on self-esteem. Higher manifest-anxiety scores were found among the people who regarded their nail biting, hair-pulling, object-chewing, nervous twitching, or giggling as serious problems. Finally, people who regarded their nail-biting as more serious tended to have higher obsessive-compulsive scores. The results in general suggest that the frequency of several of these behaviors is anxiety-related and that it is the person's assessments of these behaviors as problems rather than simply their frequency that is related to higher anxiety and lower self-esteem. PMID- 8234611 TI - Loneliness and its relationship with misery. PMID- 8234612 TI - Speed and accuracy of memory decisions in older adults. AB - An experiment examined the speed and accuracy with which older individuals made decisions about the contents of their memory. 63 volunteers aged between 50 and 80 years made speeded decisions about whether they knew the meaning of rare words presented to them on a computer screen followed by a forced-choice recognition test to assess the veracity of the memory decisions. Subjects were required to select 3 targets from a list of 10 that together constituted the definition of the original word. Analysis showed (i) the older adults were slower to decide whether or not they knew the words, (ii) positive decisions were quicker when more was known about the target, (iii) the age difference in decision speed was unrelated to the amount known about each item. PMID- 8234613 TI - Animistic thinking in psychotic versus conduct-disordered hospitalized adolescents. AB - 25 adolescents showing a Psychotic-spectrum Disorder were compared with 24 adolescents showing Conduct Disorder on a task eliciting reasons for attributing life to an ambiguous pictorial stimulus (dough on a beach). Consistent with expectations from earlier research, the psychotic adolescents showed greater modes of animistic thinking (attributions of life to nonlife forms) than conduct disordered youth, associated with theorized merging between self and the nonhuman environment. Implications are discussed. PMID- 8234614 TI - Overwhelming remembrance of things past: Proust portrays limbic kindling by external stimulus--literary genius can presage neurobiological patterns of puzzling behavior. AB - Proust detailed inexplicable behavior long before the neurobiologists Goddard and McIntyre in 1972 demonstrated that intermittent repetition of harmless stimuli can cause "kindling" of a seizure (with or without motor convulsions). Such brief seizures can occur especially in the evolutionarily old limbic system which mediates basic drives, their concomitant emotions, and certain aspects of memory. It appears that in humans the influence of specific external stimuli that revive the memory of repeated past experiences may "kindle" a transient episode of limbic overactivation. Thereupon the normal balance between the limbic and frontal lobe systems is disturbed (for a few minutes) as are normal human decision making and control of action. Linked with such a transient frontolimbic imbalance is out-of-character behavior, psychosis (hallucinations or delusions), autonomic activation, and severe distortion of affect and of action, culminating in extreme cases in a "Limbic Psychotic Trigger Reaction," as proposed by Pontius in 1981, in motiveless homicidal acts with mostly preserved consciousness and memory for the acts. PMID- 8234615 TI - Tedium, depression, and suicidal preoccupation. PMID- 8234616 TI - Causal attributions and coronary heart disease in women. AB - The relationship between coronary heart-disease endpoints and attributional style in women has been previously unexamined. This study examined the attributions of 73 postmyocardial infarction (MI) women about their heart disease and explored the relationship between attributions and nonfatal coronary recurrence. Women's primary causal attributions included personal behavior (9.6%), blaming others (19.3%), stress (28.8%), luck (12.3%), and family history (13.7%). The largest proportion of recurrences occurred in women attributing their infarcts to marital problems. Of the attributional ratings, ascriptions involving spouses were the only attributions that met entry criteria for logistic regression (p = .019) after controlling for severity of first infarction. PMID- 8234617 TI - Education and suicide: the quality of life among modern Americans. PMID- 8234618 TI - Adolescent offenders: comparison for sexual, violent, and property offences. AB - The psychological functioning and abuse history of 23 sexual, 51 violent, and 79 property adolescent offenders were compared. Minnesota Multiphasic Personality Inventory scores of these three groups did not differ although sexual offenders were twice as likely to have a history of sexual abuse, making those personal experiences more relevant to the form of their criminal behavior. PMID- 8234619 TI - Onsite psychological evaluation of a hostage taker. AB - Many law enforcement agencies have responded to the challenge of dealing with hostage situations by developing negotiation teams that include psychological consultants. These consultants are useful in determining whether the hostage taker is mentally ill and suggesting possible strategies for negotiation. The traditional psychological evaluation must be modified to fit the unique stresses and requirements of a hostage situation. Suggested is an onsite assessment that includes a mental status evaluation based on observation of a client and a selective history taken from collateral sources. PMID- 8234620 TI - Confusing the terms "guilty" and "not guilty": implications for alleged offenders with mental retardation. AB - Evaluations of competency to stand trial of 45 alleged offenders with mental retardation were inspected to examine their understanding of "guilty" and "not guilty." Seven defendants (16%) had virtually no knowledge of "guilty" and were judged not competent after a thorough evaluation. None received a jail sentence. Although the criminal justice system served well this group with mental retardation, questions remain about the fate of those not identified. Are some pleading guilty when actually meaning "I did not do it" and being surprised by an unexpected jail sentence? PMID- 8234621 TI - Divorce and dating violence revisited: multivariate analyses using Straus's conflict tactics subscores. AB - 514 men and 891 women college students provided information concerning behaviors both they and their partners used within the prior six months to resolve conflicts in their relationships. Multivariate analyses assessed whether experiencing the divorce of one's parents would be associated with respondents' report of their own or their partners' conflict behaviors. Students from divorced families reported higher scores for their own behavior on the Violence subscale only, while they reported higher scores for their partners on both the Verbal Aggression and Violence subscales. These results suggest that coming from a divorced family may have lasting effects on later relationships of these individuals, particularly in conflict resolution. PMID- 8234622 TI - Language in differential diagnosis of a teenage schizophrenic girl with aspects of multiple personality. AB - Diagnosis of a teenage schizophrenic girl showed many features of her language and behavior mimicked the presentation of a Dissociative Personality Disorder. These aspects were secondary to the primary schizophrenic disorder and treated via a multimodal approach combining pharmacotherapy, individual, family, and milieu therapy. Implications are drawn for tailoring verbal interventions to such overdetermined conditions. PMID- 8234623 TI - Eating attitudes test in boys and girls aged 6-18 years: decrease in concerns with eating in boys and the increase in girls with their ages. AB - Concerns with eating were studied in 130 Japanese boys and 125 girls aged 6-18 years using the Simplified Eating Attitudes Test (s-EAT). The s-EAT scores in girls slightly increased with age. The mean scores in girls at age 10 years or older were significantly higher than in boys of the same age, suggesting that pubertal girls have more concerns with eating. On the other hand, s-EAT scores in boys that were not overweight decreased as they grew older, contributing, at least partly, to the sexual difference in eating behavior. The mean scores in overweight boys were higher than in boys that were not overweight. The score in boys correlated significantly with weight though there was no significant correlation in girls. These results suggest that, in addition to increased concerns with eating in girls, decreased concerns with age in boys is one of the causes of the sexual difference in eating behavior, especially during puberty. Eating behaviors in girls seem to be less influenced by changes in body weight than in boys. PMID- 8234624 TI - Neuropsychological prediction of treatment efficacy and one-year outcome in schizophrenia. AB - The present study was conducted to establish the degree of interrelationship between neuropsychological functioning in the acute phase of the schizophrenic illness, clinical measures of treatment response (positive and negative symptoms), and 1-year outcome. Nineteen SADS/RDC schizophrenic inpatients were clinically rated during a 2-week drug washout period and again following 4 weeks of neuroleptic treatment. The findings revealed that reduced attentional ability in the baseline phase significantly predicted the presence of higher residual negative symptoms following 4 weeks of treatment, while neurocognitive status was unrelated to positive symptom response. With respect to prediction of the 1-year outcome, poor functional status of this schizophrenic population was significantly associated with the presence of baseline memory dysfunction. PMID- 8234625 TI - Somatopsychic aspects of paranoia. AB - The factors to which the pathogenesis of paranoia is ascribed are predisposition, personality and environment. This paper deals with further etiological preconditions such as psychoorganic impairment, convulsive disorders and physical handicaps together with their psychoreactive processing, based on detailed case reports. Such patients are relatively frequent within the (epidemiologically small) group comprising paranoia. The psychodynamic factors underlying the sensitive processing of physical handicaps are described, with means of prevention being discussed in conclusion. PMID- 8234626 TI - Clinical features of situational and nonsituational major depression. AB - Clinical lore has long supported the subtype of situational or reactive depression. To date, however, there has been limited empirical research support for this subtype of major depression. We examined demographic, clinical and personality features of situational and nonsituational depression in 89 outpatients with unipolar nonpsychotic major depressive disorder. Situational depressives had a less recurrent course of illness and appeared to respond more completely to the antidepressant used for their current episode. Demographic and personality measures did not distinguish situational and nonsituational depression. PMID- 8234627 TI - Obsessive-compulsive disorder and depression. A retrospective study on course and interaction. AB - In a retrospective study 62 patients, who fulfilled the ICD-8/9 criteria for obsessive-compulsive disorder (OCD), were followed up. Besides an assessment of the cross-sectional symptomatology of OCD and depression, the long-term course of OCD and its relationship to depression were investigated. Five courses of OCD could be differentiated: continuous and unchanging (27.4%); continuous with deterioration (9.7%); continuous with improvement (24.4%); episodic with partial remission (24.2%), and episodic with full remission (11.3%). There was no difference between primary or secondary depression on the prognosis of OCD, and there was also no difference between the continuous or episodic course with regard to primary or secondary depression. Our results may be biased by the fact that we selected a sample of OCD and not primarily major depressive patients. PMID- 8234628 TI - Comparison of schizophrenic delusions between Japan and Germany. AB - The content of schizophrenic delusions of 150 German patients in the Psychiatrische Universitatsklinik Tubingen and those of 324 Japanese patients (ICD-9:295) were compared according to the classification of Huber and Gross. The comparison revealed significantly higher frequencies of delusions of poisoning (18 vs. 8%; p < 0.01) and jealousy (6 vs. 1.9%; p < 0.05) in the German cases, but there was no significant difference in the incidence of comprehensive injury delusions. There was also no significant difference in the incidence of comprehensive grandeur delusions. However, comprehensive belittlement delusions were more frequent in the German group (24 vs. 13.3%; p < 0.01). Among the delusions of this type, the themes of guilt/sin (15.3 vs. 4.9%; p < 0.001), particularly those related to religion, were more frequent in the German group. Although religious delusions were more frequent in the German group (21.3 vs. 6.8%; p < 0.001), the difference regarding the incidence of 'delusions of world destruction' was not statistically significant. Among the persecution/injury delusions, themes of direct persecution from others (Verfolgungswahn) were conspicuous in the German group, whereas delusions of reference (Beziehungswahn) related to harassment, such as 'being slandered by others' or 'being known', were common in the Japanese group. Namely, the difference in the content of these persecution/injury delusions may derive from the different types of self in the two countries: the individually oriented self in Germany and the group-oriented self in Japan. Delusions of guilt/sin in the German group may reflect the characteristics of sin in Christianity. PMID- 8234629 TI - Influencing and being influenced: the other side of 'bizarre delusions'. 1. Analysis of the concept. AB - The psychopathological validity of the current concept 'bizarre delusion' is questioned. A review of the literature reveals that the traditional category 'ego disorders'--though preferable to 'bizarre delusion'--is also biased, taking into account mainly 'passivity experiences' of the kind of 'Gemacht', eclipsing experiences of active influence on external reality. Although anecdotal records of activity experiences in schizophrenic psychoses have been reported since Kraepelin, any general systematization is lacking. PMID- 8234630 TI - Influencing and being influenced: the other side of 'bizarre delusions'. 2. Clinical investigation. AB - A clinical follow-up lasting from 1 to 6 years on 25 outpatients affected by endogenous psychoses and manifesting ego disorders demonstrates the presence of both activity and passivity experiences in schizophrenias and affective psychoses. No differential diagnosis between the two nosographical groups can be based on the phenomenological characteristics of passivity and activity experiences. Regarding the schizophrenic sample, our observations suggest reconsidering on theoretical grounds the validity of the construct 'loss of ego boundaries'. PMID- 8234631 TI - Abnormal illness behaviour and somatisation due to leucorrhoea. AB - The belief that passage of a 'whitish discharge' is associated with bodily complaints of weakness, tiredness, exhaustion, multiple aches, and multiple somatic complaints is known to be widely prevalent among Asian women. However, this aspect has not gained research attention. The case reported here illustrates how multiple somatic complaints, psychosomatic in nature, are attributed to the passage of white discharge and manifest as an abnormal illness behaviour. PMID- 8234632 TI - Stability of diagnoses in various psychiatric disorders: a study of long-term course. AB - The frequency of diagnostic shift was investigated in 267 patients diagnosed with various psychiatric disorders according to the 9th revision of the International Classification of Diseases (ICD-9). Forty-six patients fulfilled the diagnostic criteria for schizophrenic psychosis, 71 for affective psychosis, 66 for neurotic disorder, 24 for personality disorder, and 40 for psychogenic reaction. The remaining 20 patients were diagnosed with heterogenous disorders. The mean follow up time was 12.5 years. Every episode of inpatient treatment was diagnosed cross sectionally. Patients with an initial episode of schizophrenic psychosis showed by far the greatest stability (93%). A high stability was found in patients diagnosed with neurotic disorders (79%). Forty-two patients with an index diagnosis of affective psychosis suffered more than one type of episode during the course of disease. The lowest stability was noted in patients with a psychogenic reaction (10%). Eleven percent of patients with a final diagnosis of schizophrenia had an index diagnosis of psychogenic reaction. Our findings demonstrate that the course of disease should be considered in making the final diagnosis. PMID- 8234633 TI - Delusions of transformation of the self. AB - A series of 20 patients suffering from delusions of physical and psychological transformation of the self is studied. Nosological, psychosocial, and biological aspects are discussed. Two cases are presented in detail. PMID- 8234634 TI - Some remarks on dysphoria from an anthropological point of view. AB - The concept of 'dysphoria', as defined by the Vienna Group from the psychopathological point of view, is analyzed on the anthropological level. In an anthropological perspective, personality is conceived as a factor modulating affective disorders, and the manifold clinical expressions of affective disorders are viewed as functions of the relationship between the entity of the 'endothymic' fluctuations and the amalgam of the person who is in charge to confront and cope with the thymic experience. The hypothesis that dysphoria may be interpreted as a sthenic, oppositional and therefore irritated and hostile personological reaction to the occurrence of endogenous 'restrictive' or 'expansive' mood swings is discussed. PMID- 8234635 TI - Conceptual disorganization in children at risk for schizophrenia. AB - The present study investigated the presence of conceptual disorganization among children at risk for future emotional disturbance. 105 children born to schizophrenic, depressed, and unaffected mothers were administered the Object Sorting Test (OST) which was scored for both developmental quality and the gradations of deviant responses. The results indicated that children with schizophrenic mothers exhibited more conceptual disorganization and lower scores for developmental quality in concept formation on the OST than did children of either depressed or unaffected mothers. For the three groups of children, lower scores for developmental quality in concept formation were significantly associated with ratings of greater severity in overall behavior dysfunction. PMID- 8234636 TI - Postpartum psychoses: onset and long-term course. AB - 86 cases of narrowly defined postpartum psychosis were investigated with regard to onset and course of symptomatology. The most frequent initial symptoms were restlessness, paranoid symptoms, catatonic excitement, anxiety, sleep disturbances and depressed mood. 16 patients had died at the time of follow-up. Concerning the further course of the disease 64% of the 61 women who were followed up (average 26 years from onset), had recurrences of illness. The most frequent longitudinal diagnosis, in 49% of the 61 cases, was 'schizoaffective disorder'; only 28% of patients were diagnosed as having schizophrenic disorders at follow-up. PMID- 8234637 TI - The modality of reconstruction in 'immodithymia'. AB - We studied the modality of reconstruction in the depressive patients whose premorbid personality presents 'immodithymia'. The patients try to pursue the reconstruction limitlessly in obsessive ways. That is, there is no discharge from their reconstruction, and the limitless succession of reconstruction supports the patients' existence. The hindrance of working for limitless succession provides the soil from which depression arises. The limitless reconstruction reflects an aspect of the deterioration of the depressive existence that is the loss of temporality or the dehistoricalization. On the other hand, when we consider the basic structure of manic-depressive psychosis under the concept of the excessive identification of the inner attitude, we can position the deterioration of our patients' existence between that of melancholics and that of manics. PMID- 8234638 TI - Factors influencing the long-term outcome of schizoaffective disorders. AB - Factors influencing the long-term outcome of schizoaffective disorders were investigated in 101 patients with a mean duration of illness of 25.5 years. Univariate statistical analyses and stepwise multiple discriminance analyses were applied. The most important factors influencing the development of persisting psychosocial alterations proved to be the absence of pure melancholic episodes during course and life events at onset, and the presence of first rank schizophrenic symptoms, schizomanic-depressive mixed episodes, higher number of episodes and higher annual frequency of episodes. The most relevant factors influencing the development of negative social consequences due to the illness were found to be an asthenic/low self-confident premorbid personality and the higher number of episodes and cycles. There are only partial similarities with schizophrenic or affective disorders. PMID- 8234639 TI - Age disorientation in Kraepelinian schizophrenia: frequency and clinical correlates. AB - Many schizophrenic patients exhibit significant neuropsychological impairment, and age disorientation is considered to be one of the more extreme manifestations. To evaluate clinical correlates of age disorientation with reference to the course of schizophrenic illness, we compared 39 deteriorated chronic Kraepelinian schizophrenic patients and 39 nondeteriorated schizophrenic patients early in the course of illness. Age disorientation was observed in 36% of patients in the Kraepelinian group, but in no patient in the non-Kraepelinian group. In the Kraepelinian group, age disorientation was unrelated to positive/negative symptoms or illness duration. Our data suggest that age disorientation may be a function of aging in schizophrenia, but is not merely a function of chronicity. PMID- 8234640 TI - The influence of social factors on the response to stress. AB - The studies described in this paper provide evidence for the importance of social factors in determining the response of the hypothalamic-pituitary-adrenal (H-P-A) system to stress. The presence of familiar social partners can reduce or eliminate the cortisol response to either loss of a significant social relationship or to fear inducing stimulus. In contrast, disruptions of ongoing social behavior can be a potent activator of the H-P-A axis. Social isolation, the formation of new social groups of unfamiliar monkeys, and disruption of social behavior under conditions of high demand food seeking, all result in elevations of circulating cortisol. Of importance is that these elevation persist for protracted periods of time and represents an animal model of chronic stress. PMID- 8234641 TI - Endocrine correlates of stress vulnerability. AB - Psychological stress is implicated as one element in a constellation of risk factors for the development and maintenance of psychosomatic disease. Endocrine responses to stressful experience vary widely among individuals, and may account for some of the individual variability in vulnerability to development of psychosomatic symptoms under stress. Endocrine stress responsiveness probably has a large genetic component, but is also known to be modifiable by prolonged and/or profound stressful experience. Social and cognitive factors influence the pituitary-adrenal response to stress, and sex differences may also be significant. It may be possible to develop specific tests of endocrine responsiveness, perhaps incorporating psychological and pharmacological challenges, that may serve as markers of chronic stress experience or of vulnerability to psychosomatic disorders. PMID- 8234642 TI - A clinical study of nightguard vital bleaching. AB - This double-blind, randomized, controlled clinical trial was designed to evaluate the efficacy and soft tissue effects of nightguard (mouthguard) vital bleaching. Subjects were assigned to one of two treatment groups, overnight (insertion of mouthguard at bedtime) or 3-hour (replenishment of test solution at hourly intervals for a total of 3 hours exposure daily). The experimental solutions included two products that are specifically designed for tooth bleaching, one product that is marketed as an oral antiseptic, and a control solution (glycerin). Each subject participated for a 3-week period. Tooth color analysis (shade determination) was done at baseline and the end of the treatment. Gingival and Plaque Indices were recorded at baseline and at the end of each week of study. Results indicated that the bleaching treatment was effective in most cases and caused no tissue inflammation or significant tooth sensitivity. The mean Gingival and Plaque Indices were generally lower (denoting improvement) at the end of treatment. Patient satisfaction with the procedure was high. PMID- 8234643 TI - Bonded composite resin Class II restoration: an example. AB - Bonded composite resin can be used for restoration of certain teeth with posterior proximal carious lesions. Successful restoration depends on careful tooth isolation, proper dentin and enamel bonding methods, selection of an appropriate composite resin, and ideal handling of all the restorative materials. This paper details a procedure for placing a mesio-occlusodistal bonded composite resin restoration in a maxillary premolar. PMID- 8234644 TI - Precision of hand-held dental radiometers. AB - The intensity of light within the wavelengths of 400 to 500 nm on a dental photocurable resin restorative material is a vital factor affecting many of the physical and chemical properties of the resulting restoration. This paper compares the precision of two recently introduced hand-held radiometers. The results indicate that the response of both hand-held meters was linear. For a given amount of decrease in light source intensity, both hand-held units responded with a corresponding decrement in measured intensity value. Both hand held meters were found to limit their pass band of intensity readings between 400 and 515 nm, making them very useful clinical instruments. However, the absolute intensity readings between the two units were significantly different. PMID- 8234645 TI - A clinical study of an intracoronal fixed partial denture attachment system. AB - An alternative tooth replacement system, based on pin retention, was clinically evaluated for up to 4 years in a prospective study. Thirty-two Universal Dental Anchorage anchors, supporting 28 fixed partial prostheses in 25 patients, were placed. Most of these prostheses had a cast complete crown on one abutment; on the other abutment, the UDA anchor functioned as a support for the extension pontine. After a mean observation period of 2 years, nine anchors failed. Around five of the failed anchors, caries was present; this occurred after 2 years or more of function of the prostheses. Early failure (4/9) were registered in the first year of function and were caused by loss of retention (three) or anchor fracture (one). The periodontal condition of the UDA anchor abutment teeth was not significantly different from that of the contralateral side. The number of failures, especially the number caused by recurrent caries, is cause for concern. PMID- 8234646 TI - Effect of luting media on the compressive strengths of two types of all-ceramic crown. AB - This study evaluated the effect of selected luting media on the compressive strength of two types of all-ceramic crown. Tooth preparation was standardized; each preparation had a shoulder width of approximately 1.2 mm, and all internal preparation angles were rounded. Hi-Ceram and Dicor all-ceramic crowns were fabricated and cemented into the preparations with zinc phosphate, glass-ionomer, or composite resin cement. Coronal compressive fracture strengths were determined, using a set of unrestored teeth as a control. There were no statistically significant differences among the mean compressive strengths of the three luting media, and there was no statistically significant difference between the mean compressive strength of Dicor and that of the natural tooth control. PMID- 8234647 TI - Self-reported occupation-related health complaints among dental laboratory technicians. AB - Occupation-related health complaints were investigated among 101 women and 100 men employed in dental technology in Norway. In technicians older than 30 years, there were no age-related differences in frequency of self-reported health complaints. Gender-related differences in responses were limited to specific musculoskeletal symptoms. About half of the employees had experienced some kind of job-related health problem, among which musculoskeletal and dermal reactions were common (39% and 28%). Other health problems were respiratory (16%), systemic (16%), sensory (10%), or neurologic (7%). Ergonomics and work-specific stress factors were important etiologic factors for the musculoskeletal reactions, whereas chemical factors of dental material origin were related to the other reactions. Comparison of the present results with corresponding data from other occupations indicated that the duties of a dental technician combine the risk of ergonomically induced musculoskeletal reactions characteristic of manual labor with the risk of chemically induced reactions. PMID- 8234648 TI - An analysis of causes of apical root resorption in patients not treated orthodontically. AB - With few exceptions, most cases of external apical root resorption have no evident etiology, particularly when root blunting from orthodontic treatment has been ruled out. This study determined the frequency of apical root resorption in the permanent dentition of patients who had not been treated orthodontically. Associations between the occurrence of external apical root resorption and three measures of oral health--number of missing teeth, periodontal probing depths, and alveolar crestal bone heights--were also examined. Between 7% and 10% of the 306 patients exhibited obvious apical resorption. Strong statistical associations were found between the occurrence of external apical root resorption and loss of teeth, increased periodontal probing depths, and reduced crestal bone heights; ie, root resorption was significantly more common in teeth with compromised support. Loss of stability from adjacent teeth, increased use of fewer remaining teeth, and loss of the root's anchorage in the bone are significant predictors of external apical root resorption. PMID- 8234649 TI - Composite resin inlays: a study of marginal adaptation. AB - Ten mesio-occlusodistal composite resin inlays were fabricated by the indirect method and cemented in extracted posterior teeth. The restorations were subjected to thermocycling and subsequently placed in a 50% silver nitrate solution. Specimens were prepared and inspected under a stereoscopic microscope and a scanning electron microscope. The results showed a total absence of microleakage at the margins terminating in enamel and a slight degree of microleakage in only two margins terminating in cementum. This corresponded with the general absence of defects and gaps that was observed in the scanning electron microscope. The exception was in the two specimens that exhibited dye penetration; these were found to have material in the marginal areas. PMID- 8234650 TI - Criteria of applicability for autoradiography of tritium. AB - Autoradiography is an effective tool for the imaging of radionuclide distributions in various samples. In sophisticated applications with special preparation and development of sample-emulsion combinations and subsequent grain counts it can be highly quantitative, but it requires carefully controlled conditions and a variety of counter-checks, for example through scintillation spectroscopy. Less refined applications use X-ray films as detectors, and their seeming simplicity tends to invite artefacts and misinterpretations. Particular care needs to be taken, if one deals, or presumes to deal, with the low-energy beta-emitter tritium. Because of the short electron ranges the film must be in intimate contact with the sample, which tends to produce chemographic artefacts; without added spectroscopic measurements it is impossible to discriminate the spurious signals from a blackening of the film due to tritium. Recent statements concerning autoradiographic tritium measurements in tree samples have created considerable public concern and have demonstrated the pitfalls of uncritical use. This paper presents order-of-magnitude criteria for the detection threshold in the autoradiography of tritium; they can serve as an exclusion principle for some of the more extravagant misinterpretations. PMID- 8234651 TI - Environmental behaviour of radionuclides deposited after the reactor accident of Chernobyl and related exposures. AB - Several radioecological experiences with isotopes of ruthenium, iodine, caesium and barium, obtained after the reactor accident of Chernobyl, are reported. It was found that for a wet deposition barium was the element with the highest retention on grass. The retention of caesium was lower by a factor of 1.6, retention of iodine by a factor of 2.4 and retention of ruthenium by a factor of 3.5. Former data on the caesium transport in cereals from leaves to grain were confirmed. Depending on the conditions the iodine transfer factor for milk varied between 0.002 and 0.007 d.kg-1. The caesium transfer factor for milk was (0.003 +/- 0.0006) d.kg-1 and was found to be relatively constant in the years 1986 1988. In 1991, the values were higher by a factor of 2-5. Radioecological model results of whole-body burdens in Southern Bavaria showed a reduction of the caesium ingestion doses due to countermeasures and spontaneous changes of consumption habits in the first 2 months by a factor of 5 and till the end of 1987 by a factor of 1.5. The model results agree well with the range of measured whole-body burdens. One month after the deposition, the external exposures in urban environments due to ruthenium, caesium and barium isotopes were found to be reduced by a factor of 2, compared with open lawns, in the case of iodine the reduction was even a factor of 2.5. External exposures of a population group from the Munich area were determined by thermoluminescence dosimetry. The results are in accordance with the spectral measurements of external dose rates in urban environments. PMID- 8234652 TI - Effective dose--how effective for patients? AB - The question discussed in this paper is whether effective dose can reflect the risk to patients from radiological procedures and can be used, for example, to optimise procedures and compare risks of various methods, to define dose constraints, and to estimate the risks to individuals or populations attributed to medical exposures. This report demonstrates that the use of effective dose for patients could be misleading or even wrong due to inappropriate simplifications of the underlying biological mechanisms and inappropriateness of the weighting factors connected with the definition of effective dose for a given patient population. We show that the choice of the most meaningful quantities to express patient exposure depends strongly on the respective situation. PMID- 8234653 TI - Behaviour of magnetic micro-particles in the human lung. AB - Magnetic micro-particles were used to investigate the defence system of the human lungs against foreign material. After primary magnetisation a remanent magnetic field (RMF) of the lung can be measured that allows estimation of the amount of dust retained in the lung. After calibration of the system with a lung phantom the magnetic contamination retained in the lungs of dental technicians and welders was estimated at mean values of 22 and 500 mg respectively. In normal controls only 0.3 mg was found. About 0.5 mg of spherical monodisperse magnetite particles was deposited in the alveolar region of the lung by voluntary inhalation. The decay of the RMF, called relaxation, results from a misalignment of the dipole particles due to the activity of pulmonary macrophages. This macrophage activity is characterised by a cellular energy Ez. With a secondary magnetisation the lung can be remagnetised by rotation of the dipole particles. This allows an estimation of the intracellular viscoelasticity and the motility of the alveolar macrophages in vivo. Secondary magnetisation and relaxation curves of spherical monodisperse magnetic particles are presented. Intracellular viscosity was estimated to be n approximately equal to 100 Pa.s at shear rates near 0.01 s-1, the rigidity modulus being v approximately equal to 4-8 Pa. Macrophage activity was described by a cellular energy EZ approximately 5 x 10( 18) J. Additionally, non-magnetic aerosol exposure resulted in a faster relaxation, which was interpreted to be due to activation of the macrophages. The magnetite particles were cleared with a half-time of approximately 110 days. PMID- 8234654 TI - Relation between rate of cell proliferation and formation of micronuclei after combined treatment with X-rays and caffeine. AB - We studied the effects of caffeine (2 mM), X-rays (1 Gy) and the combination of both agents on cell proliferation and formation of micronuclei in the early stages of preimplantation mouse embryos in vitro. Two-cell embryos were exposed to the agents shortly before division to the 4-cell stage. Proliferation and micronucleus production was monitored every 2 h in the 4- and 8-cell stages. A rather peculiar pattern of micronucleus formation after radiation exposure alone was observed for 8-cell embryos: those embryos that were the first to enter the 8 cell stage showed two to three times higher numbers of micronuclei per cell when compared with those embryos that entered the 8-cell stage some hours later. Studies of the kinetics of cell proliferation and of micronucleus formation in 4- and 8-cell embryos and exposure to caffeine revealed that this result could be explained by two factors: a slight asynchrony in the developmental stage at the time of exposure and the length of the interval being available for repair processes. When caffeine was present, a third factor had to be taken into consideration: direct inhibition of repair by caffeine. PMID- 8234655 TI - Estimation of double-strand break quality based on track-structure calculations. AB - The aim of this study was to investigate whether double-strand break (DSB) quality is related to the complexity of radiation-induced damage or to the spatial distribution of initial DSBs. The analysis was based on track structure calculations and DNA damage modelling. The results obtained indicate that the quality of DSBs is related to their initial spatial distribution rather than to their complexity. PMID- 8234656 TI - Different rates of ultraviolet-induced DNA damage in the epidermis and dermis of a platyfish model for carcinogenesis. AB - The purpose of this study was to compare fluence-response relationships for the production of cyclobutane pyrimidine dimers in epidermal or dermal DNA of platyfish Xiphophorus hybrids irradiated with UVB, and to determine photoreactivation from black light on dimers produced in situ. This was accomplished by quantitative gel electrophoresis of unlabeled DNA following extraction of the DNA and treatment with an enzyme specific for the detection of pyrimidine dimers. The dermis was the target tissue for UV-induced DNA damage in Xiphophorus hybrid fish skin. Shapes of dimer-fluence response data following filtered sunlamp irradiation (lambda > 290 nm) or monochromatic wavelength 302 nm in the epidermis or dermis were different. In the epidermis there was an initial step upward slope followed by a plateau, whereas in the dermis a linear relationship was observed. The final values of dimers at the high doses were, however, nearly equal in the epidermis and dermis exposed to either radiation. These differences in fluence-response relationships are probably attributable to the intertwining of the epidermis and to the shielding effect of the epidermal layer, with scales leading to a heterogeneous population of cells which are exposed to different UV doses. Photoreversal of dimers was readily observed by black light irradiation in both epidermis and dermis irradiated with either lambda > 290 nm or 302 nm. PMID- 8234657 TI - Using technique factors to predict patient ESE. AB - This paper describes a formula for estimating patient skin entrance exposures from representative techniques used in radiographic facilities. A flexible nomogram has been formatted to the parameters of patient dimensions and focal film distance. The corresponding exposure rate (mGy/mAs) is expressed for any assumed projection and selected kilovoltages. This measured approach is a valuable tool for exposure assessment and is in compliance with emerging government standards and the pursuit of the ALARA (as low as reasonably achievable) concept. PMID- 8234658 TI - How breast cancer patients choose a treatment method. AB - This paper discusses two treatment options for early stage breast cancer- modified radical mastectomy and lumpectomy plus radiation therapy--and examines the reasons why women choose one method of treatment over the other. In examining how women make their treatment decision, the paper reviews the literature regarding how psychological, emotional and psychosocial factors play a role in determining a method of treatment. It also discusses coping methods during breast cancer treatment, why women choose to undergo breast reconstruction and to whom they look for information when making health decisions. PMID- 8234659 TI - Understanding quality control. AB - Most of us have trouble defining "quality," but we all know what competency means. Rules, regulations and guidelines have been established to ensure competency among the members of our profession. Quality control is a common, easy way to demonstrate competency. It is not aimed at individuals; it is aimed at processes. Our goal, then, should be to develop QC programs that allow us to demonstrate that we are competent in the operation of our business--the production of consistently acceptable radiographs. PMID- 8234660 TI - Wiktor stent may offer new promise. PMID- 8234661 TI - The baccalaureate degree in radiography. PMID- 8234662 TI - In Hong Kong, change is a way of life. PMID- 8234663 TI - How JRCERT handles student complaints. PMID- 8234664 TI - RCEEMs, recorders, developers, sponsors. PMID- 8234665 TI - When a mammogram alone isn't enough. PMID- 8234666 TI - Just another flash in the pan. PMID- 8234667 TI - What makes an occupation a profession? PMID- 8234668 TI - [Function-related morphology of the intervertebral disks]. AB - The intervertebral discs contribute to shock absorption only to a limited degree; they are important however, for efficient orientation of axial pressure transmission to the entire surface of the contiguous vertebral bodies. So long as there is sufficient distension of the nuclei pulposi, the annuli fibrosi (particularly those of the lumbar vertebral column) play a part in the ligamentous control of the motion segment. From the 20th year of life onwards the discs are completely avascular, although they show high metabolic turnover. Their water content decreases with advancing age. As early as the first decade of life, beginning in the lateral, uncovertebral zones, clefts appear in the cervical intervertebral discs. These clefts frequently progress to tears involving the whole of the intervertebral disc. PMID- 8234669 TI - [Modern intervertebral disk diagnosis]. AB - Recent advances in our knowledge of the pathophysiology and function of the intervertebral disc, in combination with the use of CT and MRI have changed radiological diagnostic algorithms and enlarged our diagnostic scope. Microfissures in the cartilage endplates of the disc and in the endplates of the vertebral bodies and anulus fibrosus are the sequelae of loss of hydration of the nucleus pulposus leading to instability and abnormal load stress. MRI shows these early degenerative changes in the disc, but also reactive processes in the disc and the neighbouring vertebral bodies. On conventional X-rays only segmental malfunctions are visualized. Protrusion and prolapse can be recognized very well by CT, but best of all by MRI. Myelography and discography are very invasive and rarely used. Very important is that protrusion and prolapse are very often diagnosed without any clinical symptoms and that both may heal without any treatment. Therefore conservative treatment of prolapse and/or protrusion is the management of choice. Blastoma and infection of the disc are found in the vast majority of cases only in combination with corresponding diseases of the vertebral bodies. The only important exception are small children, in whom primary bacterial infection of the disc is well known. PMID- 8234670 TI - [Diagnostic imaging following surgery of spinal disk herniation]. AB - The magnetic resonance imaging findings recorded in patients after surgery for degenerative disc disease in the lumbar and cervical spine are discussed in comparison with conventional radiographs and computed tomography findings. In the lumbar spine normal postoperative findings in the immediate postoperative period can be demonstrated by MR imaging. Contrast-enhanced MR imaging can differentiate disc herniation from postoperative scar formation with a greater degree of confidence than other imaging modalities. MR imaging improves differentiation between other causes of failed back syndrome such as postoperative hematoma and infection, lateral spinal stenosis and arachnoiditis. In the cervical spine types of operative approaches, the appearance of bony stenosis and disc herniations by MR imaging are discussed. Computed tomography still has a role in the assessment of osseous complications such as central or foraminal stenosis. PMID- 8234671 TI - [Percutaneous nucleotomy]. AB - Percutaneous nucleotomy (PN) is a non-operative, percutaneous, intervening, radiologically controlled method for the treatment of non-sequestrated herniated discs. The diagnostic criteria are CT, MR, functional myelography with myelo-CT, discography and disco-CT. Of the indication for this procedure is strictly confined to patients pretreated in the conventional way, this method yields good or very good results in about 75-80% of cases. PN is performed under local anesthesia and under fluoroscopic control. Of the technical parameters are strictly observed, PN is considered a low-risk method. The rate of infection is lower than 1%. Even though there are no long-term results yet, the positive results we have observed for 1 to 6 years suggest that this method will become an established method of treating herniated discs, as there is little discomfort for the patient. PMID- 8234672 TI - [Cerebral manifestations of progressive facial hemiatrophy (Romberg's disease). Nuclear spin tomographic findings and literature review]. AB - Progressive facial hemiatrophy (PFH) is a sporadic disease of unknown etiology. It is characterized by shrinking and deformation of one side of the face. Potential CNS involvement has repeatedly been suspected in some patients, but is still a matter of controversy. In this article we describe the clinical and MR imaging findings of the CNS in three female patients with PFH and present a comprehensive review of the literature. One of three PFH patients had partial epilepsy. MRI showed ventricular enlargement, white matter lesions, flattening of the cortical surface and meningeal adhesions homolateral to the facial hemiatrophy. Two other patients had completely normal intracranial findings. These findings confirm that cerebral hemiatrophy occurs in a subgroup of PFH patients. The MRI pattern, however, does not seem to be consistent with a simple atrophic or malnutritional process. We consider chronic localized meningoencephalitis with vascular involvement as a possible underlying mechanism for the occasional CNS involvement in PFH. PMID- 8234673 TI - [An unclear osteolysis of the right lower jaw. Histiocytosis X]. PMID- 8234674 TI - [An unclear bone swelling? Osteogenesis imperfecta cystica]. PMID- 8234675 TI - [The radiologic diagnosis of venous diseases. A challenge]. AB - Limited accuracy in the clinical diagnosis of deep vein thrombosis (VT) makes such diagnostic tests such as duplex sonography or venography necessary. Exact information on the age and extent of the thrombus are necessary for the clinician to optimize the therapeutic management. The correct diagnosis of calf vein thrombosis and of recurrent VT in patients with postphlebitis changes also has implications for treatment. After exclusion of thrombosis, the radiologist should evaluate the leg for other possible causes of symptoms besides VT. Investigation of the venous system also has a role in the diagnosis in patients with suspected pulmonary embolism. In patients with chronic venous insufficiency the deep venous system should be assessed for patency and venous valve function. The superficial veins should be differentiated in segments with sufficient or insufficient venous valves, and it is also necessary to look for insufficiency of the perforating veins. In patients with superficial phlebitis there is risk of propagation into the deep venous system. PMID- 8234676 TI - [Imaging procedures in varicosities]. AB - A prospective study was performed to compare the diagnostic potential of color coded duplex sonography (CCDS) and that of ascending pressure phlebography (APP) in 61 patients before venous stripping. Varicosis of the long saphenous vein was correctly diagnosed and graded by CCDS in 87% of cases, and varicosis of the short saphenous vein in 97%. CCDS was inaccurate in 3 of 4 cases of recurrent varicosis of the long saphenous vein. Anterior femoral cutaneous varicosis (n = 3), mild postthrombotic syndrome (n = 1) and variant venous anatomy (n = 1) were not diagnosed on CCDS. One Baker cyst was diagnosed by CCDS. In the assessment of venous reflux CCDS seems to be superior to APP. Our results suggest that CCDS is a valuable diagnostic tool for routine imaging before venous stripping. However, patients with ambiguous or inconclusive CCDS results (eg. recurrent varicosis, venous variants) should be examined by APP. PMID- 8234677 TI - [Phlebography of the leg veins. Study quality and sources of error]. AB - In a retrospective study, 198 contrast venographies were evaluated by two blinded observers for quality of opacification of the venous system and quality of documentation. There was good correlation between the two observers in 91% of the cases. They found an adequate contrast quality in 183 cases (92.4%), while in 15 cases (7.6%) incomplete contrast was found in some parts of the calf or pelvic veins. Most venograms (195, or 98.5%) were adequately documented. In conclusion, contrast venography is an excellent method for evaluation of the venous system, if performed adequately. Nevertheless, there is room for improvement of both technique and documentation. PMID- 8234678 TI - [A comparison of sonography and phlebography in the diagnosis of deep venous thrombosis of the lower extremities]. AB - In recent years, non-invasive sonographic techniques [real-time ultrasound (US), duplex ultrasonography (DU) and color-coded duplex sonography (CCDS)] have increased in importance for the diagnosis of lower extremity deep venous thrombosis. To ascertain the accuracy and limitations of these techniques prospectively, various studies have been performed to compare sonography with venography. This review deals with basic methodologic considerations when two imaging methods are compared and describes the features of deep venous thrombosis seen on US, DU and CCDS. Reports comparing sonography and phlebography for the diagnosis of deep venous thrombosis are presented and discussed. The advantages and limitations of US, DU and CCDS for the diagnosis of iliac, femoral, popliteal and infrapopliteal venous thrombosis are compared with those of venography, and controversial areas are touched on. PMID- 8234679 TI - [The value of spiral-CT in primary and secondary liver neoplasms]. AB - Sixty-one patients in whom primary or secondary malignomas of the liver were suspected on the basis of ultrasound examination were examined with spiral CT and sequential CT during i.v. administration of contrast medium. In four further cases spiral CT-angiography (spiral-CTA) was performed. Advantages and disadvantages of spiral volume CT are described. PMID- 8234680 TI - [Methodology, technical prerequisites and postoperative morbidity of intraoperative radiotherapy (IORT) of soft tissue sarcomas. Heidelberg Krankengut 6/91-9/92]. AB - Since June 1991 the IORT facility has operated a dedicated linear accelerator, which was installed within the central operating theater of the Department of Surgery. As of 9/92 a total of 28 patients suffering from peripheral (n = 20) or centrally (n = 8) located soft tissue sarcomas had been were treated. Thirteen patients revealed a primary and 15 patients a recurrent tumor. Tumor resection with negative margins was performed in 20 patients, positive margins remained in 5 patients, and gross macroscopic residual disease in 3 patients. Combined intraoperative and external beam radiotherapy was applied in 22 patients, using IORT doses of 10-20 Gy and an external beam dose of 26-50 Gy. Three patients were irradiated intraoperatively twice with a time interval of 24 h. After a median follow-up of 9.9 months, 20 patients are disease free. Two patients died 4 and 5 months after the end of therapy with rapidly progressive distant metastases. An infield failure within the external beam target volume was seen in 1 patient and local failure at the field margin of the external field in 3 patients. So far, there have been no IORT infield failures. Follow-up is performed with magnetic resonance imaging. In 3 patients a second operation was necessary because of a severe wound infection, including one patient suffering from osteomyelitis of a neighboring bone. Mild sensory neuropathy occurred in 1 patient 7 months after treatment. Overall only mild and reversible postoperative and posttherapeutic complications were seen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234681 TI - [Soft tissue calcification in the back of the knee]. PMID- 8234682 TI - [36th Annual Meeting of the Hessen Association of Medical Radiation Therapy together with the South-West German Association of Radiologists and Nuclear Medicine Physicians. Bad Nauheim, 24-26 September 1993. Abstracts]. PMID- 8234683 TI - Epilepsy: surgery and imaging. PMID- 8234684 TI - Spiral (helical) CT. AB - Spiral (helical) computed tomography (CT) involves continuous patient translation during x-ray source rotation and data acquisition. As a result, a volume data set is obtained in a relatively short period of time. For chest or abdominal scanning, an entire examination can be completed in a single breath hold of the patient or in several successive short breath holds. The data volume may be viewed as conventional transaxial images or with multiplanar and three dimensional methods. The authors review the technologic aspects of spiral CT, as well as its advantages, limitations, and current clinical applications. PMID- 8234685 TI - Dynamic MR imaging of the head and neck: an idea whose time has come ... and gone? PMID- 8234686 TI - A critical appraisal of the Canadian National Breast Cancer Screening Study. PMID- 8234687 TI - Review of stereotaxic large-core needle biopsy and surgical biopsy results in nonpalpable breast lesions. PMID- 8234688 TI - Atypical hyperplasia: frequency and mammographic and pathologic relationships in excisional biopsies guided with mammography and clinical examination. AB - PURPOSE: To ascertain the frequency and pathologic relationships of atypical hyperplasia in biopsy specimens obtained after clinical and mammographic examination. MATERIALS AND METHODS: Clinical, mammographic, and histologic findings were prospectively correlated in 300 consecutive excisional biopsies. RESULTS: Atypical hyperplasia was detected in 26 (17%) of 154 biopsies with benign findings and 19 (13%) of 146 biopsies with malignant findings overall (P > .05). The frequency of atypical hyperplasia was only 4% (two of 55 specimens) in clinically prompted biopsies with benign findings but 24% (24 of 99 specimens) in mammographically prompted biopsies with benign findings (P = .002), increasing to 31% (21 of 68 specimens) in benign microcalcifications and 40% (18 of 45 specimens) in benign microcalcifications associated with adenosis. Atypical hyperplasia was found most often within (16 [62%] of 26 cases) or adjacent to (nine [35%] of 26 cases) another lesion that prompted biopsy. CONCLUSION: Most atypical hyperplasia is not a random finding in benign biopsy specimens but shows a statistically significantly greater frequency in association with microcalcifications detected with mammography only, especially those in areas of adenosis. PMID- 8234689 TI - Breast cancer malpractice litigation in New York State. AB - PURPOSE: To identify causes of purported malpractice in diagnosis and treatment of breast cancer. MATERIALS AND METHODS: The authors reviewed cases from The New York Jury Verdict Reporter listed between 1985 and 1991 to look for those in which there was alleged delay in diagnosis or treatment of breast cancer. RESULTS: Of 34 cases identified, 32 (94%) were based on presumed delay in diagnosis and only two (6%) on claims of therapeutic malpractice. Delay in diagnosis was commonly claimed in patients younger than 50 years (76%). Palpable masses were present in 94% of these cases. Either mammograms were not obtained (16 cases, 50%) or findings were interpreted as normal or as fibrocystic disease (12 cases, 38%). Specialists most frequently cited were gynecologists (16 of 39,41%). Highest awards (> or = $1 million) were more commonly given to patients younger than 50 years with proved distant or nodal metastasis (six of 34, 18%). CONCLUSION: Emphasis on early diagnosis has led to the perception that purported delay in diagnosis, however short, even in the presence of a palpable mass, changes the chances for survival. PMID- 8234690 TI - Hemoptysis: comparative study of the role of CT and fiberoptic bronchoscopy. AB - PURPOSE: A prospective study was performed to compare results of computed tomography (CT) and fiberoptic bronchoscopy in diagnosis of cancer in patients with hemoptysis. MATERIALS AND METHODS: Ninety-one patients (64 men, 27 women) with hemoptysis underwent thin-section CT and fiberoptic bronchoscopy. RESULTS: CT scans demonstrated all 27 tumors seen at bronchoscopy and an additional seven, five of which were beyond bronchoscopic range. In patients with normal chest radiographs, bronchial carcinoma was detected in 5% at both bronchoscopy and CT. In patients with abnormal findings on radiographs, bronchoscopy allowed both location and histologic diagnosis in 78% of carcinomas but was unreliable in locating peripheral tumors demonstrated at CT. CT was insensitive in demonstrating early mucosal abnormalities, bronchitis, squamous metaplasia, and a benign papilloma, all detected at bronchoscopy. CONCLUSION: The initial examination should be bronchoscopy when there is high clinical suspicion of carcinoma and relevant radiographic abnormality, and CT when strong clinical suspicion of carcinoma is not substantiated at bronchoscopy in patients with normal findings on chest radiographs. PMID- 8234691 TI - End-stage lung disease: CT findings in 61 patients. AB - PURPOSE: To determine whether the cause of end-stage lung disease could be determined on the basis of pattern and distribution of abnormalities at computed tomography (CT). MATERIALS AND METHODS: The CT scans from 61 consecutive patients with end-stage lung disease were independently assessed by two observers without knowledge of clinical or pathologic data. End-stage lung disease was defined by the presence of honeycombing, extensive cystic changes, or conglomerate fibrosis. The observers listed the three most likely diagnoses and recorded the degree of confidence in the first-choice diagnosis on a three-point scale. RESULTS: The observers made a correct first-choice diagnosis in an average of 87% of cases. A correct first-choice diagnosis was made most often in silicosis (100%), Langerhans cell histiocytosis (100%), asbestosis (90%), usual interstitial pneumonia (88%), extrinsic allergic alveolitis (87%), and sarcoidosis (83%). There was good agreement between the two observers on the first-choice diagnosis (kappa statistic, 0.89). CONCLUSION: The pattern and distribution of abnormalities in end-stage lung disease are determined by the underlying cause. In most cases, a specific diagnosis can be made on the basis of the CT findings. PMID- 8234692 TI - Importance of ground-glass attenuation in chronic diffuse infiltrative lung disease: pathologic-CT correlation. AB - PURPOSE: To correlate areas of ground-glass attenuation at computed tomography (CT) with findings at open lung biopsy in chronic diffuse lung disease. MATERIALS AND METHODS: The cases of 26 patients were included on the basis of (a) extensive areas of ground-glass attenuation as the predominant (n = 17) or exclusive (n = 9) abnormality at CT in the absence of honeycombing and (b) histologic evaluation at open lung biopsy. Severity of ground-glass attenuation was scored in the lobe sampled at biopsy, with separate analysis of associated lung changes. RESULTS: Correlation of CT with histologic findings at the 37 biopsy sites demonstrated that ground-glass attenuation corresponded to inflammation in 24 (65%) cases and to fibrosis in 13 (54%) cases. Eleven of the 13 patients (85%) with fibrosis had traction bronchiectasis or bronchiolectasis. These findings were not present in any of the patients with inflammation. CONCLUSION: In patients with chronic diffuse infiltrative lung disease, areas of ground-glass attenuation not associated with traction bronchiectasis or bronchiolectasis are a reliable indicator of inflammation. PMID- 8234693 TI - Quantitative differential pulmonary perfusion: MR imaging versus radionuclide lung scanning. AB - PURPOSE: To compare the results of quantitative analysis of differential pulmonary artery blood flow performed with phase-contrast cine magnetic resonance (MR) images with those of differential radionuclide pulmonary perfusion scanning in preoperative lung transplantation patients. MATERIALS AND METHODS: Forty preoperative lung transplantation patients underwent prospective, quantitative analysis of differential pulmonary artery blood flow performed with MR imaging and radionuclide pulmonary perfusion scanning. MR imaging analysis was performed at 1.5 T with a phase-contrast cine pulse sequence to acquire data perpendicular to each central pulmonary artery. RESULTS: Comparative analysis of differential pulmonary perfusion showed excellent correlation between the two modalities in all patients (P = .0001). MR imaging enabled calculation of the pulmonary artery blood flow to each lung; radionuclide lung scanning enabled only calculation of relative perfusion to each lung. CONCLUSION: MR imaging is as effective as radionuclide lung scanning in assessment of relative, and superior for determination of absolute differential, pulmonary perfusion. PMID- 8234694 TI - Anterior mediastinal lesions: transsternal biopsy with CT guidance. Work in progress. AB - PURPOSE: To assess the technique, results, and patient tolerance of transsternal biopsy of anterior mediastinal lesions under computed tomographic (CT) guidance. MATERIALS AND METHODS: Seven patients (four women and three men) with anterior mediastinal lesions underwent eight transsternal biopsies. Patients were included in the study if there was a risk of injury to internal mammary vessels or a danger of tranversing the lung by the biopsy needle. Patients were specifically questioned about the pain caused by the transsternal needle. A pain scale was not used. Conscious sedation and local anesthesia requirements provided gross evaluation of the patients' tolerance to the procedure. CT scans were used to guide and confirm optimal alignment of the transsternal needle with the lesion. RESULTS: Patients experienced minimal discomfort when the sternum was traversed. No instances of pneumothorax, postprocedural pain, or infection were encountered. In six of the seven patients, biopsy specimens were diagnostic. CONCLUSION: The transsternal approach for biopsy of anterior mediastinal lesions appears safe and is well tolerated. PMID- 8234695 TI - Hyperintense uterine leiomyoma at T2-weighted MR imaging: differentiation with dynamic enhanced MR imaging and clinical implications. AB - PURPOSE: To evaluate the role of dynamic enhanced magnetic resonance (MR) imaging in differentiating leiomyomas with high signal intensity at T2-weighted imaging. MATERIALS AND METHODS: Histologic features of 34 uterine leiomyomas that showed hyperintensity on T2-weighted images were compared with those of ordinary hypointense leiomyomas (n = 74). The results of treatment with gonadotropin releasing hormone analogue were also analyzed in relation to pretreatment signal intensity. RESULTS: Degenerated leiomyomas (n = 13) had higher signal intensity and heterogeneous architecture, but there was considerable overlap in signal intensity with cellular leiomyomas (n = 21). At dynamic enhanced MR imaging, cellular leiomyomas had marked contrast enhancement in the early phase, while degenerated leiomyomas showed slight or irregular enhancement. Cellular leiomyomas were reduced significantly in size after treatment (P = .02). The response in degenerated leiomyomas was minimal. CONCLUSION: Evaluation of signal intensity at T2-weighted imaging and degree of contrast enhancement at dynamic enhanced MR imaging provide useful information on the nature of leiomyomas. PMID- 8234696 TI - Decidual cyst: endovaginal sonographic sign of ectopic pregnancy. AB - PURPOSE: To determine the prevalence of decidual cysts in patients with ectopic pregnancy and assess the value of endovaginal sonographic demonstration of decidual cysts in predicting ectopic pregnancy. MATERIALS AND METHODS: A series of 288 proved ectopic pregnancies was reviewed, and a series of 179 patients with pregnancies of less than 8 weeks menstrual age was prospectively examined. RESULTS: Decidual cysts were identified in 30 (14.4%) of 208 ectopic pregnancies. In 12 patients a decidual cyst was the first sonographic sign of ectopic pregnancy, and in six patients it was the only abnormal sonographic finding. Four of five patients with decidual cysts had an ectopic pregnancy. Decidual cysts had a sensitivity of 21%, specificity of 92%, positive predictive value of 80%, and negative predictive value of 42% in the diagnosis of ectopic pregnancy. CONCLUSION: Patients with decidual cysts are at high risk for ectopic pregnancy and should be monitored or treated appropriately, depending on the clinical findings. PMID- 8234697 TI - Prenatal compensatory renal growth: documentation with US. AB - PURPOSE: To determine whether compensatory renal growth (CRG), which occurs in adults, children, and infants after loss of functioning renal tissue, occurs in fetal life. MATERIALS AND METHODS: Prenatal ultrasound scans were used to select 29 patients with a unilateral multicystic dysplastic kidney and four patients with unilateral renal agenesis. Accurate measurements of prenatal renal length were obtained in 21 of these patients; an accurate measurement of postnatal renal length and birth weight was obtained in 27 of these patients. Prenatal and postnatal renal measurements of the contralateral kidney were compared with renal length in 23 matched control patients and previously reported normal renal lengths. RESULTS: The single functioning kidneys in the study patients were significantly longer than those in the control patients before and after birth (P = .001). CONCLUSION: This study provides strong evidence to support the belief that CRG occurs before birth. PMID- 8234698 TI - Mediastinal Hodgkin disease in children: response to therapy. AB - PURPOSE: To determine the frequency of thymic and nodal enlargement at computed tomography (CT) in pediatric patients with Hodgkin disease, time to resolution of mediastinal disease, frequency of residual masses, and ability to distinguish between thymic hyperplasia and residual fibrosis with CT. MATERIALS AND METHODS: CT scans of 60 pediatric patients with Hodgkin disease were reviewed. Diagnosis of a residual or recurrent mass was based on surgical, gallium-67 scintigraphic, and clinical findings. RESULTS: In 18 (78%) of 23 patients who underwent serial chest CT, the mediastinum reverted to normal within a mean of 6.3 months. Of these 18 patients, four developed recurrent mediastinal widening due to thymic hyperplasia. Five patients had residual mediastinal masses. Benign fibrosis was confirmed in two patients at biopsy and presumed in three patients on the basis of Ga-67 scintigraphic findings. Recurrent lymphoma was not identified. CONCLUSION: Mediastinal masses resolve in the majority of pediatric patients with Hodgkin disease. Residual mediastinal widening is indicative of fibrosis, while an enlarged thymus at follow-up more likely represents hyperplasia rather than recurrent disease. PMID- 8234699 TI - Perforation complicating balloon dilation of esophageal strictures in infants and children. AB - PURPOSE: The authors retrospectively reviewed their experience with balloon dilation of esophageal strictures in infants and children to determine the prevalence of esophageal perforation during this procedure. MATERIALS AND METHODS: Balloon dilation was performed in 32 patients with esophageal stricture due to either congenital or postoperative stenosis. The total number of procedures was 141; each patient underwent one to 12 procedures. RESULTS: Four cases of esophageal perforation occurred during balloon dilation and were detected immediately after the procedure. Perforation in three of these patients was treated nonoperatively with parenteral fluid and antibiotics, and one patient was treated surgically with resection and creation of an anastomosis. The clinical course after treatment was favorable in all cases. CONCLUSION: Although esophageal perforation during balloon dilation of benign strictures has been reported as a rare complication, four perforations were observed in 32 patients. Esophageal perforation can occur in infants and children more frequently than previously reported. PMID- 8234700 TI - Cost containment in the use of low-osmolar contrast agents: effect of guidelines, monitoring, and feedback mechanisms. AB - PURPOSE: To describe a program for controlling intravenous use of low-osmolar contrast agents (LOCAs). MATERIALS AND METHODS: The department of radiology at the authors' institution adopted a policy of selective use of intravenously administered LOCAs. Clinical indications for LOCA use were specified after consultation with the administration, risk managers, legal department, and ethics committee of the hospital. The guidelines were then publicized throughout the department and hospital, and a contrast agent data form was developed to collect data on all cases. Monitoring mechanisms were instituted. RESULTS: Over the next 23 months, 11,373 patients received intravenous iodinated contrast agents, of whom 28.1% were deemed to be at high risk and were given LOCAs. Monthly tracking showed no evidence of a trend toward increasing use of LOCAs. CONCLUSION: Clear definition of use guidelines, close monitoring, and feedback can stabilize LOCA use at acceptably low levels. PMID- 8234701 TI - Oxygenation in the rabbit myocardium: assessment with susceptibility-dependent MR imaging. AB - PURPOSE: To determine the feasibility of using hemoglobin (Hb) desaturation as an indicator of myocardial oxygenation. MATERIALS AND METHODS: High-resolution gradient-echo nuclear magnetic resonance (MR) images of isolated, blood-perfused rabbit hearts were obtained at various blood oxygenation levels. The hearts were perfused at 37 degrees C with a Langendorff apparatus modified for nuclear MR imaging. The perfusate contained bovine red blood cells in a cardioplegic solution that eliminated motion artifacts and minimized arteriovenous oxygenation differences. Hb saturation was varied (7%-100%) randomly. Perfusion pressure was continuously monitored, and blood samples were obtained. RESULTS: There was a substantial correlation between image signal intensity in the myocardium and Hb saturation in the blood, believed to be due to susceptibility effects of the paramagnetic species deoxyhemoglobin. CONCLUSION: Direct and noninvasive determination of regional Hb saturation with susceptibility-dependent MR imaging may provide information regarding regional myocardial O2 content. PMID- 8234702 TI - Comparative evaluation of clinically available inferior vena cava filters with an in vitro physiologic simulation of the vena cava. AB - PURPOSE: A physiologic in vitro model of the human inferior vena cava (IVC) was developed to evaluate the clot-trapping efficiency of various IVC filters. MATERIALS AND METHODS: The flow model closely simulates the physical parameters of the human IVC in fluid viscosity, specific gravity, temperature, and pulsatility; blood flow velocity, volume, and inherent turbulence; and vein orientation, wall compliance, and clot composition. Five filters--Greenfield (G), Bird's Nest (BN), Vena Tech (VT), Simon nitinol (SN), and titanium Greenfield (TG)--were compared by using two vena cava and blood clot sizes and horizontal and vertical orientation of the model. RESULTS: Each filter varied significantly in its clot-capturing efficiency, depending on vena cava size and orientation and emboli size. Overall, decreasing rank order in filter clot-trapping efficiency was SN, BN, VT, TG, and G. CONCLUSION: The in vitro model of the human IVC is an alternative to multicenter clinical trials in evaluating the relative clot trapping efficiency of various IVC filters. PMID- 8234703 TI - Nitinol intravascular stent: results of preclinical evaluation. AB - PURPOSE: To test an expandable nitinol intraluminal stent for hoop strength, biocompatibility, corrosion resistance, and patency. MATERIALS AND METHODS: Forty four stents were implanted in the iliac arteries of 22 sheep. Follow-up was performed with angiography and histologic examination for up to 6 months. RESULTS: All but one stent remained widely patent during the follow-up period. Two stents in two sheep were placed incorrectly early in the study; these sheep were not followed up. Hoop strength was found to be superior to that of a similar, commercially available, self-expanding stent. Minimal corrosion was seen at 6 months, and the stent appears to be biocompatible. The stent has good radiopacity and deploys with minimal foreshortening. CONCLUSION: The stent can be reliably and safely deployed in the vascular system. Clinical trials in humans are warranted. PMID- 8234704 TI - Renal artery stenosis: analysis of Doppler waveform parameters and tardus-parvus pattern. AB - PURPOSE: The utility of Doppler parameters and waveform contour analysis for diagnosis of renal artery stenosis was studied in a hypertensive population screened for renovascular hypertension. MATERIALS AND METHODS: The Doppler results were compared with those obtained with angiography in a prospective double-blind study involving 118 kidneys in 46 patients with hypertension and 11 potential renal donors with normal blood pressure. Doppler parameters measured in the distal renal artery included systolic acceleration, acceleration time, peak systolic velocity, deceleration in late systole, pulsatility and resistivity indexes, and waveform features in early systole. RESULTS: Angiograms demonstrated 28 stenotic renal arteries. There was no significant difference between stenotic (> 50% diameter narrowing) and nonstenotic renal arteries for any parameters studied. When stenosis was further categorized as moderate (50%-79%) or severe (80%-95%), significant (P < .05) differences for acceleration time and systolic acceleration were found between nonstenotic and severely stenotic arteries. CONCLUSION: Doppler characterization of the tardus-parvus phenomenon in the distal renal artery is not an adequate screening method for detection of renal artery stenosis. PMID- 8234705 TI - Transjugular intrahepatic portosystemic shunt: US assessment with maximum flow velocity. AB - PURPOSE: To evaluate the ability of Doppler ultrasonography (US) to depict the patency of a transjugular intrahepatic portosystemic shunt (TIPS). MATERIALS AND METHODS: Twenty-eight patients were followed up after creation of TIPS with US, angiography, and endoscopy performed at regular intervals. Magnetic resonance (MR) angiography was performed in four patients. US was performed first, and the results were correlated with those at angiography. Measurements of maximum flow velocity (Vmax) were taken from the proximal part of the shunt. RESULTS: In normal shunts, mean Vmax was 95 cm/sec. Shunt obstruction was seen in 12 cases: occlusion in four and stenosis in eight. All cases of occlusion were detected with US and appeared as absent flow within the shunt. There was one false positive diagnosis of occlusion. All cases of stenosis resulted in reduced Vmax (mean, 32 cm/sec; P < .001). After successful revision in seven patients, Vmax rose to a mean of 122 cm/sec. Artifact from the metal stent prevented flow assessment with MR imaging. CONCLUSION: US can be performed to evaluate shunt status, and Vmax is an accurate noninvasive parameter for assessing shunt patency. PMID- 8234706 TI - Refractory ascites: early experience in treatment with transjugular intrahepatic portosystemic shunt. AB - PURPOSE: To evaluate the transjugular intrahepatic portosystemic shunt (TIPS) in treatment of refractory ascites. MATERIALS AND METHODS: Fourteen patients with chronic liver disease and portal hypertension were included in a prospective study. Six patients had Child-Pugh class B disease; eight had class C disease. Indications for TIPS were three previous hospital admissions over 9 months for treatment of tense ascites, no response to diuretic therapy, or an occluded peritoneovenous shunt with tense ascites. The volume of ascitic fluid was sonographically evaluated before and after TIPS placement. RESULTS: The technical success rate for TIPS creation was 93%. Mean portosystemic gradient decreased from 22.8 mm Hg +/- 7.2 before TIPS placement to 11.3 mm Hg +/- 3.6 after TIPS (P = .005). Complete resolution of ascites was achieved in seven (50%) patients. Treatment failed in seven; five had Child-Pugh class C disease, and four of these had a Child-Pugh score greater than 11. CONCLUSION: These results are comparable to those for other forms of treatment of ascites. The authors do not, however, recommend TIPS in patients with a Child-Pugh score greater than 11. PMID- 8234707 TI - Mechanical thrombectomy: results of early experience. AB - PURPOSE: To evaluate the clinical application of a mechanical thrombectomy device in occluded lower extremity arteries and bypass grafts. MATERIALS AND METHODS: A mechanical thrombectomy device was used in five patients with acutely occluded lower extremity bypass grafts (n = 2) or superficial femoral arteries (n = 3). RESULTS: The thrombus was completely removed in three cases (two bypass grafts and one native superficial femoral artery). In two of the patients with occlusions in the native superficial femoral artery, the thrombus was partially removed. In these two patients, the aspirate demonstrated a more fibrous thrombus that was not easily fragmented. In one of these two patients, the coagulum was thought to be embolic from the heart. CONCLUSION: The thrombectomy device works well in recent thromboses, particularly in polytetrafluoroethylene grafts in which organization does not occur. Its efficacy in chronically occluded vessels is not certain. PMID- 8234708 TI - Change induced by radiation therapy in FDG uptake in normal and malignant structures of the head and neck: quantitation with PET. AB - PURPOSE: To quantitate the changes induced in uptake of the glucose analog 2 [fluorine-18]fluoro-2-deoxy-D-glucose (FDG) in normal structures in the head and neck and compare these to the change in uptake in malignant structures in patients with head and neck tumors undergoing radiation therapy. MATERIALS AND METHODS: Eleven patients with biopsy-confirmed squamous cell carcinoma of the head and neck were studied before, during, and after a 6-week course of radiation therapy with positron emission tomography (PET)-FDG imaging. A ratio of FDG uptake in the structure compared with that in the cerebellum (termed metabolic ratio) within and outside of the field of radiation was determined in the adenoids; lingual and palatine tonsils; parotid, submandibular, and sublingual glands; and nasal turbinates, soft palate, and gingiva. RESULTS: The average metabolic ratio in the tonsils, nasal turbinates, soft palate, and gingiva did not change significantly with treatment. CONCLUSION: FDG uptake in normal structures does not change with radiation therapy. This fact is in marked contrast to the FDG uptake in squamous cell carcinomas in the head and neck, which decrease dramatically with treatment (P < .005). PMID- 8234709 TI - Dynamic MR imaging in the head and neck. AB - PURPOSE: To determine the utility of dynamic magnetic resonance (MR) imaging in evaluation of head and neck lesions. MATERIALS AND METHODS: Dynamic MR imaging was performed in 79 head and neck lesions. Five types of time versus signal intensity curves were obtained: type A, curve peaks 0-30 seconds after administration of contrast material; type B, curve peaks at 30-60 seconds; type C, curve peaks at 60-210 seconds; type D, gradual upward slope; and type E, flat. RESULTS: Use of dynamic imaging did not improve the sensitivity of nondynamic imaging for malignancy. Although malignancies had type B, C, or D curves, 39 (68%) of 57 benign lesions had the same patterns. Lesions with type A or E curves and lesions with type D curves and with marked hyperintensity on T2-weighted images were all benign. CONCLUSION: Although use of dynamic MR imaging contributes little in predicting malignancy in head and neck lesions, it can limit the differential diagnosis and has potential in predicting vascularity and recurrence. PMID- 8234710 TI - Elusive "stuck" disk in the temporomandibular joint: diagnosis with MR imaging. AB - PURPOSE: To determine the incidence of a "stuck" disk and its importance in the overall scheme of internal derangement. MATERIALS AND METHODS: Magnetic resonance (MR) images of 212 temporomandibular joints (TMJs) in 106 symptomatic patients were obtained and reviewed. Image analysis included assessment of disk position; disk mobility; and degree of condylar translation, with or without disk recapture in joints with displaced disks. RESULTS: Of the 127 joints with disks in a normal position, 94 had mobile disks and 23 had stuck disks. Ten joints revealed no condylar translation, which precluded assessment of disk mobility. Of 85 joints with displaced disks, 67 joints showed mobility, with recapture in 25 and no recapture in 42. Significant correlation existed between disk mobility and degree of condylar translation (P < .001). CONCLUSION: The TMJ disk may be stuck in a normal or displaced position and limits the normal degree of condylar translation, leading to restricted jaw opening. Recognition of a stuck TMJ disk could alter patient care. PMID- 8234711 TI - Frameless method of stereotaxic localization with DSA. AB - PURPOSE: To improve a method of stereotaxic localization with digital subtraction angiography that does not require use of a localization frame fixed to the patient's skull during examination. MATERIALS AND METHODS: An independent, low cost, stereotaxic computing system was devised. Software programs used magnetic resonance images, computed tomographic scans, and digitized radiographs of plastic bone implants with head landmarks to establish reference trihedrons in each imaging system, transpose target coordinates from one system to another, adjust surgical instruments, help plan radiation surgery, and compute and display isodose curves. RESULTS: The geometric distortions of the image intensifier were corrected. Distortion problems of the conic projection were solved, and three dimensional localization was achieved with only two associated front and lateral views. Accuracy to within 1 mm was achieved for the three coordinates. CONCLUSION: This frameless stereotaxic localization technique is highly accurate and reliable. The system allows maximum automation of examination procedures. PMID- 8234712 TI - Complex partial seizures and mesial temporal sclerosis: evaluation with fast spin echo MR imaging. AB - PURPOSE: This study was performed to determine the sensitivity and specificity of fast spin-echo (FSE) magnetic resonance (MR) imaging in detecting mesial temporal sclerosis. MATERIALS AND METHODS: Twenty-one patients with a diagnosis of complex partial seizures were studied. Criteria for abnormality included visual detection of hippocampal MR signal abnormality and atrophy or significantly decreased hippocampal volume determined by using MR morphometric techniques. RESULTS: The overall sensitivity and specificity of MR imaging in demonstrating correct lateralization of seizure focus were both 100%. The positive predictive value of FSE MR imaging was 100%. The negative predictive value was 50%. CONCLUSION: The authors recommend FSE MR imaging of the hippocampus as the imaging technique of choice in the evaluation of patients with complex partial seizures for mesial temporal sclerosis. PMID- 8234713 TI - Amyotrophic lateral sclerosis: T2 shortening in motor cortex at MR imaging. AB - PURPOSE: To determine whether decreased signal intensity of the motor cortex (T2 shortening) at magnetic resonance (MR) imaging is a useful finding for supporting the diagnosis of amyotrophic lateral sclerosis (ALS). MATERIALS AND METHODS: High field-strength (1.5-T) MR images of 15 patients (seven men and eight women, aged 28-80 years) and 49 neurologically normal age-matched control patients were examined for T2 shortening in the motor cortex. In addition, brains of patients with ALS were examined at autopsy. RESULTS: The MR images of 14 of the 15 patients showed T2 shortening in precentral cortices, while the images of all but one of the control patients showed no such finding. In three of eight brains at autopsy, sections from the precentral cortex showed sparsely distributed, intensely stained astrocytes and macrophages. CONCLUSION: Abnormal iron deposition associated with the degenerative process could be the source of T2 shortening, which is a useful MR imaging finding in the diagnosis of ALS. PMID- 8234714 TI - Standardized uptake values of normal tissues at PET with 2-[fluorine-18]-fluoro-2 deoxy-D-glucose: variations with body weight and a method for correction. AB - PURPOSE: To study the relationship between standardized uptake values (SUVs) in normal tissues and body weight determined with positron emission tomography (PET). MATERIALS AND METHODS: SUVs were determined in 28 nondiabetic women with newly diagnosed untreated primary breast cancers. Body weights ranged from 45 to 107 kg. SUVs for lung, liver, marrow, spleen, blood, and normal breast were determined from images obtained with PET 50-60 minutes after injection of 2 [fluorine-18]-fluoro-2-deoxy-D-glucose (FDG). RESULTS: There was a positive correlation between SUV and body weight for blood, (r = .75, P = .000004), liver (r = .72, P = .000081), and spleen (r = .57, P = .033), all of which had substantial F-18 activity. In heavy patients, SUVs for those normal tissues were up to twice those of lighter patients. Calculating SUV on the basis of predicted lean body mass (SUV-Lean) eliminated the weight dependence of the SUV for blood (r = .26, P = .18) and reduced weight dependence in other tissues. CONCLUSION: SUV-Lean is a weight-independent index for blood FDG accumulation at PET and may be more appropriate than SUV for quantifying F-18 activity in tumors. PMID- 8234715 TI - Small hepatocellular carcinomas: two-phase dynamic incremental CT in detection and evaluation. AB - PURPOSE: To compare dynamic incremental computed tomography (CT) with other techniques in detecting and evaluating small hepatocellular carcinomas (HCCs). MATERIALS AND METHODS: Two-phase (early vascular and parenchymal) dynamic incremental CT of the entire liver and ultrasonography (US) were performed in 184 patients with chronic liver disease. Thirty patients had 36 small HCCs (less than 30 mm in diameter); 27 of these 30 patients also underwent CT after intraarterial injection of iodized oil, and the results were compared. RESULTS: The HCC detection rate was 92% with dynamic incremental CT, 69% with US, and 81% with iodized oil-enhanced CT. Of six well-differentiated HCCs, two were detected with the first dynamic incremental CT series and two with iodized oil-enhanced CT, whereas all were detected with the second dynamic incremental CT series and five with US. CONCLUSION: Two-phase dynamic incremental CT was superior to the other techniques and is a useful and easy procedure for routine follow-up of patients with chronic liver disease. PMID- 8234716 TI - Hepatocellular carcinoma: correlation of CT, angiographic, and histopathologic findings. AB - PURPOSE: To elucidate the causes of various enhancement patterns of hepatocellular carcinomas (HCCs). MATERIALS AND METHODS: Computed tomographic (CT) scans of 68 surgically resected HCCs (62 patients) were obtained 45 seconds and 6 minutes after administration of contrast material. These scans were compared with angiographic and histopathologic findings. RESULTS: On the early images, 24 HCCs (35%) were hypoattenuating, 33 were totally or partially hyperattenuating (48%), and 11 were isoattenuating (16%) compared to adjacent liver. On the delayed images, 55 HCCs (81%) were hypoattenuating and 13 (19%) were isoattenuating. Twenty-nine of the 48 very hypervascular or hypervascular tumors (60%) were hyperattenuating or partially hyperattenuating at early CT. Sixteen of the 20 slightly hypervascular or hypovascular tumors (80%) were isoattenuating or hypoattenuating. CONCLUSION: Although there was relatively good agreement between tumor vascularity and enhancement pattern (60% correlation for hypervascular tumors, 80% correlation for hypovascular tumors), hyperattenuation of the large HCCs (> or = 5 cm) at CT appeared to be a function of dilated sinusoids within the tumor (peliotic changes) as well as vascularity. In patients with advanced cirrhosis, hypovascular HCCs could be hyperattenuating at CT. PMID- 8234717 TI - Dynamic breath-hold multiplanar spoiled gradient-recalled MR imaging with gadolinium enhancement for differentiating hepatic hemangiomas from malignancies at 1.5 T. AB - PURPOSE: To compare the enhancement patterns of focal liver lesions at dynamic breath-hold gadolinium-enhanced multiplanar spoiled gradient-recalled (SPGR) magnetic resonance (MR) imaging with T2 relaxation times in the differentiation of liver hemangiomas from malignancies. MATERIALS AND METHODS: Forty-seven patients with focal liver lesions underwent MR imaging with spin-echo and gadolinium-enhanced multiplanar SPGR techniques. T2 relaxation times and enhancement patterns were compared for accuracy in liver lesion characterization. RESULTS: Enhancement patterns allowed better characterization of liver lesions than did T2 relaxation times. Only specific patterns of contrast enhancement indicated a hemangioma. Although 18 of the 33 malignancies and 10 of the 12 hemangiomas showed progressive centripetal hyperintense enhancement, only hemangiomas filled in with hyperintense peripheral nodules. Malignancies often filled in with hyperintense thick rinds. CONCLUSION: Gadolinium-enhanced multiplanar SPGR imaging allows more accurate characterization of liver lesions than does T2 relaxation time. PMID- 8234718 TI - Confluent hepatic fibrosis in advanced cirrhosis: evaluation with MR imaging. AB - PURPOSE: To assess the usefulness of magnetic resonance (MR) imaging in diagnosis of confluent fibrosis in patients with advanced cirrhosis. MATERIALS AND METHODS: Findings on pretransplantation MR images were compared with findings on pretransplantation computed tomographic (CT) scans and posttransplantation gross appearance and histologic findings in 11 patients with confluent fibrosis. In all patients, MR imaging was performed before and after administration of gadopentetate dimeglumine. RESULTS: Nine lesions appeared wedge shaped and involved the medial segment of the left lobe, anterior segment of the right lobe, or both; two lesions consisted entirely of lobar or segmental fibrosis. Associated volume loss in the affected regions was seen in 10 lesions. All 11 lesions were hypoattenuating on non-contrast material-enhanced CT scans. CONCLUSION: Findings on MR images corresponded well to CT findings and gross appearance. MR imaging provided useful morphologic information about confluent fibrosis, but the MR signal characteristics are not unique and do not enable differentiation from hepatic neoplasms. PMID- 8234719 TI - Carcinoma of the gallbladder: CT evaluation of lymphatic spread. AB - PURPOSE: To clarify the computed tomographic (CT) characteristics of positive nodes and to assess the ability of CT to demonstrate lymphatic spread. MATERIALS AND METHODS: CT findings of lymph nodes in 51 patients with histologically proved gallbladder carcinoma were correlated with pathologic findings retrospectively. RESULTS: Thirty-one of 51 patients (61%) had a total of 157 positive nodes, 60 (38%) of which were depicted at CT. A positive node was defined at CT as a soft tissue mass with an anteroposterior dimension of at least 10 mm that showed ringlike or heterogeneous enhancement. These criteria had a positive predictive value of 75% for cystic nodes, 90% for pericholedochal nodes, 100% for posterosuperior pancreatoduodenal nodes, 100% for retroportal nodes, 88% for right celiac nodes, 100% for hepatic nodes, 100% for superior mesenteric nodes, and 83% for interaortocaval nodes. CONCLUSION: CT does not reliably demonstrate all regional nodes of gallbladder carcinoma, but the criteria of size and pattern of enhancement reliably indicate lymphatic spread, which has an important bearing on disease staging and treatment planning. PMID- 8234720 TI - Recurrent rectal cancer: diagnosis with dynamic MR imaging. AB - PURPOSE: To evaluate whether dynamic magnetic resonance (MR) imaging can increase the diagnostic accuracy in suspected local recurrence of rectal carcinoma. MATERIALS AND METHODS: Eighteen patients (seven men, 11 women, aged 26-78 years) with 19 suspect lesions were examined. T1-weighted (pre- and postcontrast) and T2 weighted spin-echo images were read by three observers. Computed tissue-specific enhancement parameters were obtained and displayed on gray-scale images (pharmacokinetic mapping). RESULTS: Reading of the spin-echo images yielded a sensitivity of 91%-100% (confidence interval, 67%, 100%), a specificity of 29% 43% (12%, 67%), and an accuracy of 71%-75% (48%, 91%). Analysis of the MR data showed greater (P = .0038) and faster (P = .0018) enhancement of malignant lesions (n = 12) compared with benign lesions (n = 7). CONCLUSION: Pharmacokinetic mapping of dynamic MR imaging data allows in vivo insight into tissue physiopathology, helping differentiate benign from malignant pelvic lesions in rectal cancer. PMID- 8234721 TI - Conjugation and hybrid MR imaging. AB - PURPOSE: To evaluate the use of conjugation with hybrid magnetic resonance imaging techniques in order to reduce artifacts from inconsistencies introduced by T2 decay. MATERIALS AND METHODS: Two phantoms containing either oil or water were used to study edge spread function (ESF) and signal-to-noise ratio (S/N) for three hybrid acquisition strategies (low pass, high pass, and ramp) in which a full set was acquired or with partial acquisition and conjugation. RESULTS: With half the echoes providing coverage of the same amount of k space, signal inconsistencies were reduced by a factor of at least e. Conjugation improved ESF degradation, and sequence efficiency increased by over 50%, which compensated for a loss in S/N. CONCLUSION: The time saved with conjugation can be used to nearly double the number of sections acquired for a given repetition time (TR) or to obtain two simultaneous echoes instead of one, with reduction of artifacts. Alternatively, the imaging time can be nearly halved for a given TR and number of sections. PMID- 8234722 TI - MR imaging in the prognostication of hamstring injury. Work in progress. AB - PURPOSE: To correlate morphologic and magnetic resonance (MR) imaging findings in hamstring injury with short-term prognosis and convalescence interval (CI). MATERIALS AND METHODS: A retrospective study of 14 professional athletes with hamstring injury was performed. The injuries were categorized according to muscle group involved, percentage of cross-sectional area affected, location, and signal intensity on T1- and T2-weighted spin-echo images. RESULTS: Longer CIs (> 6 weeks) were seen in injuries with complete transection, greater than 50% cross sectional muscle involvement, ganglionlike fluid collections (long T1 and T2), hemorrhagelike signal intensity (short T1 and T2), distal myotendinous junction tears, and deep muscular tears. Shorter CIs (< 5 weeks) were seen in superficial muscle injuries and muscle belly injuries that involved small cross-sectional areas of the muscle. CONCLUSION: Prediction of CI for high-performance athletes with complete hamstring injury may be accomplished with use of MR imaging and poor prognostic factors. PMID- 8234723 TI - Bone contusions of the knee: increased lesion detection with fast spin-echo MR imaging with spectroscopic fat saturation. AB - PURPOSE: To demonstrate that T2-weighted fast spin-echo (FSE) imaging with spectroscopic fat saturation (FS-FSE) increases the conspicuity between normal marrow and bone contusions in posttraumatic knees. MATERIALS AND METHODS: Seventy six magnetic resonance (MR) studies of the knee were prospectively evaluated in 73 consecutive patients with knee pain. Conspicuity of regions of microtrabecular trauma (bone contusions) was evaluated with conventional T2-weighted imaging in the sagittal plane, T2-weighted FSE imaging in the coronal plane, T2-weighted FS FSE imaging in the sagittal plane, and conventional T1-weighted imaging in the sagittal plane. RESULTS: Twenty-six foci of bone contusion were identified in 21 knees with the FS-FSE technique. Only 16 foci were demonstrated with conventional T2-weighted imaging. Six of the sites of bone contusion were not demonstrated with the FSE technique alone (without fat saturation). Four of the bone contusions were not seen on T1-weighted images; these sites of bone contusion were substantially more conspicuous on the FS-FSE images. In addition, FS-FSE imaging was more sensitive in demonstrating the extent of microtrabecular trauma. CONCLUSION: T2-weighted FS-FSE imaging is a sensitive and rapid method of identifying and assessing the extent of microtrabecular trauma about the knee. PMID- 8234724 TI - Prior lateral patellar dislocation: MR imaging findings. AB - PURPOSE: To illustrate the constellation of magnetic resonance (MR) findings that suggest prior patellar dislocation. MATERIALS AND METHODS: A computer search of all MR examinations of the knee with a coded diagnosis of patellar dislocation performed between February 1988 and October 1990 disclosed 22 cases (11 male and 11 female patients, aged 1-70 years). RESULTS: Eighteen cases (82%) demonstrated hemarthrosis, medical retinacular disruption, and contusion of the lateral femoral condyle. In 11 cases, the patella was displaced from the trochlear sulcus, but in no patient was there complete dislocation at the time of imaging. Axial short-inversion-time inversion-recovery (STIR) images demonstrated subtle subchondral and cancellous signal intensity abnormalities that were often poorly appreciated on T2-weighted images. In 11 cases, patellar dislocation was not suspected before MR imaging. CONCLUSION: Hemarthrosis, medial patellar and lateral femoral contusion, and retinacular disruption suggest prior lateral patellar dislocation. Because of its sensitivity to subtle intraosseous signal abnormality, axial STIR imaging is a valuable adjunct in the evaluation of the acutely injured knee. PMID- 8234725 TI - MR mammographic localization. Work in progress. AB - The authors present phantom test results for a stereotaxic device that may permit simple, rapid, and accurate needle biopsy and localization of breast lesions detected at contrast material-enhanced magnetic resonance (MR) mammography. The mechanical accuracy of this prototype MR breast localizer is approximately plus or minus 3.5 mm at 5 cm. PMID- 8234726 TI - Temporal lobes: surface MR coil phased-array imaging. AB - The authors describe a four-coil surface phased-array coil for magnetic resonance (MR) imaging of the temporal lobes. On the basis of measurements in a phantom, the signal-to-noise ratio of the array is 1.67 times or more better than that obtained with a standard quadrature head coil in the region of the hippocampi. The higher sensitivity of this phased array permits use of imaging pulse sequences for which the head coil is inadequate. PMID- 8234727 TI - Radiographic film fog artifact caused by fluorescent bandage. AB - Film fog is a common cause of x-ray film artifact. The authors report an unusual source of film fogging in the darkroom--seen in three consecutive radiographs of two different patients--the exposure of film to the low light emission of a fluorescent commercial adhesive bandage worn by the technologist. Heightened awareness of the effects of these bandages will help avoid creation of this film fog artifact. PMID- 8234728 TI - Large-caliber (24-28-F) catheters for radiologically guided percutaneous procedures. AB - A catheter set originally designed as a system for placement of large thoracostomy tubes with the Seldinger technique and sequential coaxial dilations was used to place 31 catheters in 28 patients. Either computed tomographic (CT) (n = 25) or fluoroscopic (n = 3) guidance was used. The tubes were 28 F (n = 13) and 24 F (n = 18). The tubes were successfully placed in 27 of 28 patients (96%), with the infection processes completely resolved in 22 of 25 patients (88%). Use of the system is simple and safe. These large catheters may be useful for cure of processes that do not respond to drainage with smaller tubes. PMID- 8234729 TI - Dangers of acronymophilia. PMID- 8234730 TI - Stereotaxic aspiration biopsy of the breast. PMID- 8234731 TI - [Evaluation of the genetic quality of mice used in biomedical research]. PMID- 8234732 TI - [Chlamydia pneumoniae: clinical, epidemiological and laboratory aspects]. PMID- 8234733 TI - [Effect of water activity on the growth of the xerophilic mold Eurotium herbariorum]. AB - The influence of water activity (aw) on growth of a xerophilic mold isolated from dried figs and identified as Eurotium herbariorum was studied on culture media of aw adjusted with sucrose or glycerol. Rate of radial growth (kr) and lag period were the kinetic parameters analyzed. Fungal growth was inhibited at aw > 0.97. In the presence of sucrose, optimum growth was found in M6OY agar (malt agar with yeast extract and 60% W/V of sucrose, aw = 0.95). On glycerol (aw ranging from 0.65 to 0.90) the fungus did not grow at aw < 0.80. Sucrose supported better growth than glycerol at aw 0.90. PMID- 8234734 TI - [In vitro control of Sclerotinia sclerotiorum and Gaeumannomyces graminis by bacteria of the fluorescent Pseudomonas group]. AB - Thirty six fluorescent Pseudomonas isolates were obtained from the rhizosphere of sunflower plants. By antibiosis tests, the six more efficient strains in Sclerotinia sclerotiorum growth inhibition, were selected. Simultaneously, twenty three fluorescent Pseudomonas isolates were recuperated from the rhizosphere of wheat plants and the five most efficient strains in growth inhibition of the fungi Gaeumannomyces graminis were selected. The strains selected from the rhizosphere of sunflower plants had no antagonistic effect on G. graminis and the bacteria isolated from the wheat rhizosphere showed no fungistatic activity on S. sclerotiorum. These results suggest the existence of a certain degree of plant bacteria pathogenic specificity. Among the selected bacteria, the strain FF5 of P. fluorescens originated the major inhibiting halo in vitro against S. sclerotiorum (Figure 1). In liquid culture medium this bacterium produces an antifungal substance that promotes lysis of fungi mycelium (Figure 2) and inhibition of ascospore germination and is not inhibited by the presence of Fe+3 in the culture medium (Table 1). Its synthesis is not associated with the production of fluorescein. Its action is not enzymatic because it is a substance of low molecular weight (< 2000), resistant to autoclave sterilization and photo stable. The amount of NH4+ and the high pH values produced by the FF5 strain in the liquid culture medium (Table 2) are not responsible for the antifungalal action. PMID- 8234735 TI - Effect of potassium sorbate and pH on the survival of Staphylococcus aureus in saturated sugar solutions. AB - Addition of 0.3% (w/w) of potassium sorbate (a permitted food additive) coupled with a reduction of pH from 7 to 6 or 5.5, resulted in a dramatic decrease in the survival of two S. aureus strains inoculated (10(8) c.f.u./ml) in laboratory media supplemented with sugar (227 g sugar/100 g water) and incubated at 35 degrees C. These in vitro results may be of potential value to improve the efficacy of sugar therapy for the treatment of certain infected wounds; however, clinical trials are needed to confirm these findings. PMID- 8234736 TI - [Post-epizootic activity of Western equine encephalitis virus in Argentina]. AB - It is shown here the WEE virus activity in ARgentina in 1983-1986 post-epizootic period. A surveillance system was established by the equine case notification and the sentinel animal method. Among the thirteen equine focus notified between September 1983-September 1985 in Cordoba and Santa Fe Provinces, 5 presumptive cases out of 16 sick horses were confirmed by the hemagglutination inhibition test for WEE epizootic virus. Twenty eight notified human cases were studied with negative results. The neutralizing antibody (Ac NT) prevalence among sentinel horses in Cordoba Province (4%) was lower than that of Santa Fe Province (13%) in April, 1984. It was shown that Ac NT disappeared in a 12-month period in 40% of the Santa Fe horses. Two seroconversions in Cordoba (12/1983 to 4/1986) and 9 seroconversions in Santa Fe (2/1984 to 7/1986) were registered. One of the sentinel horses form Cordoba Province also seroconverted with the enzootic WEE AG 80-646 virus. Due to the recognized cross-antigenicity between both viruses, reactivity was attributed to WEE epizootic virus. No viral activity was detected in the domestic birds. With this contribution, the study of one epizootic cycle of WEE virus was completed: pre-epizootic silence (1977-1980), epizootic (1982 1983) and residual focus plus inapparent infections during a post-epizootic period (1983-1986). PMID- 8234737 TI - Responses to speech signals in the normal and pathological peripheral auditory system. AB - The responses to single (/a/ and /i/) and double vowel (/a,i/) stimuli of normal guinea pig cochlear nerve fibres are compared with those from animals with a cochlear hearing loss. When the threshold losses are sufficient to exclude the higher harmonics of the /i/, the temporal representation of the second and higher formants is lost. Smaller threshold elevations allow a representation of the second formant when the vowel /i/ is presented alone. However, under double vowel stimulation wider auditory filters allow the capture of the synchrony of high characteristic frequency fibres by lower frequencies thereby losing the higher formants of the /i/ and also much of the information about its fundamental frequency. PMID- 8234738 TI - Functional consequences of neonatal unilateral cochlear removal. AB - The physiological consequences of unilateral cochlear removal in infancy were assessed by recording the responses of neurones in the ferret inferior colliculus and superior colliculus to acoustic stimulation of the intact ear. Animals were lesioned between postnatal days P5 and P40 and survived for at least a year prior to recording. In the inferior colliculus ipsilateral to the intact ear, neurones had lower thresholds and wider dynamic ranges following earlier (P5) than following later (P40, adult) cochlear removal. In the superior colliculus contralateral to the intact ear, neurones had broader spatial tuning in response to high-level, free-field stimulation following cochlear removal at P25, than had neurones in normal, unlesioned adults. The neural map of auditory space was also disrupted in the lesioned animals. However, at low stimulus levels the auditory space map was unaffected by the cochlear removal. These results show a developmental sensitive period for the effects of unilateral cochlear removal on the responses of ferret inferior colliculus neurones, and a level-dependent effect of cochlear removal on the responses of superior colliculus neurones. PMID- 8234739 TI - Cochlear function reflected in mammalian hair cell responses. AB - This report describes response patterns recorded in inner and outer hair cells in the apical three turns of the guinea pig cochlea. Characteristic frequencies (CF) in these regions are approximately 270 Hz in turn four, 1000 Hz in turn three and 4000 Hz in turn two. Although the two receptor types exhibit differences in resting membrane potentials and in response phase at low stimulus frequencies, they both produce ac and dc receptor potentials in response to sound. When measured around CF, both cell types produce a depolarizing dc response at low and moderate levels. This contrasts with results published for basal-turn outer hair cells (Russell and Sellick, 1983; Cody and Russell, 1987) whose responses become asymmetrical only at very high levels. PMID- 8234740 TI - Functional organization and learning-related plasticity in auditory cortex of the Mongolian gerbil. PMID- 8234741 TI - New aspects of human muscle coordination as revealed by motor-unit studies. AB - Increasing evidence has become available against the view that individual muscles should be regarded as functional units in the control of force by the motor apparatus. The relative contribution of various motoneurone pools (belonging to a single muscle) to muscle force appears to depend on the direction of torque exerted by the corresponding limb. This clearly indicates the presence of more than one control or activation parameter for the total motoneurone pool of a single muscle. Windhorst et al. (1989) have even proposed a combined sensory and neuromuscular partitioning, such that segmental control mechanisms are based on subdivisions of motoneurone pool--muscle complexes. Size-related recruitment occurs within these subdivisions rather than in the motoneurone pool as a whole. Moreover, motoneurones can operate in various modes (different relative contributions of the recruitment mechanism and the firing frequency mechanism to muscle force) related either to movement or to force. This indicates that muscle coordination is organised differently in force tasks and in movement tasks. These properties of motoneurone activation mentioned above are difficult to incorporate in existing models on sensorimotor organisation. Some of these discrepancies will be reviewed in order to highlight future directions for modelling. PMID- 8234742 TI - Interactions between pathways controlling posture and gait at the level of spinal interneurones in the cat. AB - The properties of three interneuronal populations controlling posture and locomotion are briefly reviewed. These are interneurones mediating reciprocal inhibition of antagonistic muscles and interneurones in pathways from secondary muscle spindle afferents to ipsilateral and contralateral motoneurones, respectively. It will be shown that these interneurones subserve a variety of movements, with functionally specialized subpopulations being selected under different conditions. Mechanisms for gating the activity of these neurones appear to be specific for each of them but to act in concert. Interneurones which are active during locomotion and postural reactions are distributed over many segments of the spinal cord and over several of Rexed's laminae, both in the intermediate zone and in the ventral horn (Berkinblit et al., 1978; Bayev et al., 1979; Schor et al., 1986; Yates et al., 1989). The location of neurones discharging during neck and labyrinthine reflexes is illustrated in Fig. 1A and B but indications that neurones with an even wider distribution contribute to locomotion, scratching and the related postural reactions have been provided by neuronal markers which preferentially label active neurones (WGA-HRP; see Noga et al., 1987) or neurones with active genetic transcription (c-fos; I. Barajon, personal communication; Dai et al., 1991). Such a wide distribution indicates a high degree of non-homogeneity, since neurones of different functional types are usually located in different laminae. It has been demonstrated that some of these neurones may be particularly important for setting up the rhythm of muscle contractions specific for different gaits or scratching, as part of their "pattern generators" (see, e.g., Grillner, 1981). Other neurones may be primarily involved in initiation of these movements or in postural adjustments combined with them. A considerable proportion of neurones mediating these movements are nevertheless likely to be used not in one particular type of movement but in a variety of movements, and contribute to postural reactions and locomotion as well as to various segmental reflexes and centrally initiated movements; they are likely to operate as last order (premotor) interneurones of several spinal pathways to motoneurones. One of the indications that this is the case is the overlap between the areas of location of interneurones active during postural reactions, locomotion, or scratching and the areas of location of premotor interneurones (Fig. 1C,D). The latter were labelled by loading motoneurones with WGA-HRP and by its subsequent retrograde transneuronal transport (see Harrison et al., 1986).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234743 TI - Fusimotor control of proprioceptive feedback during locomotion and balancing: can simple lessons be learned for artificial control of gait? AB - The possibilities for central control of primary spindle afferents through fusimotor efferents for gain control in motor control mechanisms are briefly reviewed. While the existence of separate pathways for independent control of static and dynamic gamma-motoneurones is well established, it proved more difficult to demonstrate that gain control of spindle feedback, attributable to alterations in static and dynamic fusimotor drive, indeed took place in voluntary movements. However, earlier qualitative indications, that Ia sensitivity (and hence the balance of static over dynamic drive) was adjusted differently in different motor tasks, have recently been confirmed in experimental simulation studies, in which the fusimotor activation profiles, that were required to reproduce chronically recorded spindle Ia discharge patterns, were reconstructed. These studies indicated that Ia sensitivity and dynamic gamma-drive were low in routine movements (walking), but that they could be dramatically increased in motor tasks which were either difficult or unfamiliar (landing from falls, balancing on narrow walk beams, adjustment to imposed disturbances). This suggested that sensitization of spindle feedback could play a significant role in motor adaptation. In line with this, studies in patients with large fibre sensory (including proprioceptive) neuropathies indicated that long-term motor deficits (affecting motor adaptation and learning) could be at least as serious as short term motor dysfunction (due to loss of reflex control). It is suggested that spindle Ia feedback may play a dual role: in addition to its contribution to short-term reflex control of posture and movement, it may also be used for optimization or maintenance of motor programs, especially if its gain is increased by significant dynamic fusimotor drive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234744 TI - Gating of reflexes in ankle muscles during human stance and gait. AB - Holding the body's centre of gravity steady represents the crucial variable for the stabilization of posture in upright stance in man. Results from two experimental approaches suggest that force-dependent receptors are required, in addition to the well-known systems involved in sway stabilization, for equilibrium control. One approach concerns bilateral leg muscle activation during stance. Unilateral or bilateral leg displacements were induced while subjects stood on a treadmill with split belts. A unilateral displacement induced a bilateral EMG response. During bilateral displacements the EMG activity was linearly summed or subtracted, depending on whether the legs were displaced in the same or opposite directions. Both legs acted in a cooperative manner: each limb affected the strength of muscle activation and the time-space behaviour of the other. This interlimb coordination is suggested to be mediated by spinal interneuronal circuits, which are themselves under supraspinal (e.g., cerebellar) control. The other approach concerns the modulation of postural reflexes under stimulated "microgravity" ir. water immersion. An approximately linear relationship was found between contact forces and impulse-directed EMG response amplitudes in the leg muscles. Out of water loading of the subjects resulted in no further increase of the response amplitude. It was concluded that the function of proprioceptive reflexes involved in the stabilization of posture depends on the presence of contact forces opposing gravity. Extensor load receptors are thought to signal changes of the projection of body's centre of mass with respect to the feet. The interaction of the afferent input from these receptors with the other systems involved in postural control is not yet fully understood. PMID- 8234745 TI - Modification of reflexes in normal and abnormal movements. AB - The trajectories observed for the limb during human locomotion are determined by a mixture of influences, some arising from neural circuits entirely within the central nervous system and others arising from a variety of sensory receptors. Muscle reflexes are highly modulated during locomotion in an adaptive manner within each phase of the step cycle. Furthermore, the modulation can be modified quickly for different tasks such as standing, walking and running, probably by changes in presynaptic inhibition. This modulation is often lost or severely reduced in patients with spasticity after spinal cord or head injury. In normal subjects cutaneous reflexes can be completely reversed from exciting to inhibiting a muscle during each step cycle, particularly in muscles that normally show two bursts of activity per cycle (e.g., tibialis anterior). In some patients stimulation of a mixed nerve (e.g., common peroneal) can directly produce muscle contraction, generate a reflex response (flexor reflex) and transiently reduce spasticity in antagonist (extensor) muscles. Thus, simple systems employing stimulation can enhance gait to a certain extent in patients with incomplete injuries. PMID- 8234746 TI - Synaptic organization of the vestibulo-collic pathways from six semicircular canals to motoneurons of different neck muscles. AB - The pattern of inputs from six semicircular canals to neck motoneurons was investigated by stimulating six ampullary nerves electrically and recording intracellular potentials from motoneurons of the rectus capitis dorsalis (RD), the complexus (COMP) and the obliquus capitis caudalis (OCA) muscles at the upper cervical cord of the cat. RD and COMP motoneurons received disynaptic excitation from bilateral anterior and contralateral horizontal ampullary nerves and disynaptic inhibition from bilateral posterior and ipsilateral horizontal ampullary nerves. OCA motoneurons received excitation from ipsilateral vertical and contralateral horizontal ampullary nerves and inhibition from contralateral vertical and ipsilateral horizontal ampullary nerves. Ipsilateral disynaptic inhibitory postsynaptic potentials and contralateral disynaptic excitatory postsynaptic potentials to these motoneurons were mediated by the medial longitudinal fasciculus (MLF) and the other postsynaptic potentials by the extra MLF pathways. The results indicated that motoneurons of a neck muscle have its own characteristic pattern of inputs from six semicircular canals. PMID- 8234747 TI - Sound preprocessing by ac and dc movements of cochlear outer hair cells. AB - In inner and outer hair cells, a sound event results mechano-electrically in a receptor potential from the hair cells by the functioning of apical and lateral K(+)-channels. However, after this point, the signal transfer is divided. Inner hair cells (IHC) release an unknown afferent transmitter. By contrast, outer hair cells (OHC) are proposed to produce mechanical ac and dc responses. In our model, the ac components of the sound signal, the carrier frequencies, determine the response of the OHC. Usually, they respond by ac and dc movements. The rapid ac movements of OHC, for which the underlying mechanism is unknown, may respond cycle-by-cycle to and interfere with the carrier frequency of the traveling wave. Near hearing threshold, they could drastically amplify the traveling wave thus contributing to the postulated cochlear amplifier. Active dc movements of the cytoskeleton of the cell body, as well as of the cuticular plate with the sensory hairs, are proposed to respond to millisecond changes of the sound stimulus over time. Such changes could be a modulation of the amplitude (AM), i.e., an increase or decrease of the sound pressure level (SPL), which is reflected in the envelope of the traveling wave. The active mechanical dc response of OHC to the amplitude (AM) and frequency modulation (FM) pattern is then expected to result in dc position changes of the reticular lamina (RL). These should control the operation point of the stereocilia, thus influencing their transfer function and sensitivity. In addition, experimental data suggest that there are modulations of the compliance of the organ of Corti (OC) and changes of its geometry. This dc modulation of micromechanical properties and geometry of the OC by active force generation of OHCs might contribute to automatic gain control, adaptation, TTS, as well as to the homeostasis of the basilar membrane location. In particular, the motile mechanism may protect the vulnerable cochlear partition against high sound pressure levels. Moreover, according to this model, changes of the sound signal with time are expected to be encoded in the actively produced dc movements of the RL. As the signal changes may carry important information (e.g., complex sound signal modulations such as formant transitions in speech), this is extracted and mechanically encoded by the proposed active dc mechanism. It cannot be excluded that the information-carrying dc signal is transferred to inner hair cells contributing to their adequate stimulus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234748 TI - Vestibulospinal reflexes and the reticular formation. AB - While both vestibulospinal and reticulospinal tracts contribute to vestibulospinal reflexes, their respective roles are not fully understood. Previous evidence suggests that reticulospinal fibers make an important contribution to the horizontal vestibulocollic reflex (VCR) of the decerebrate cat. Recent work addresses their contribution to the vertical VCR. On the basis of study of reflex and vestibulocollic neuron dynamics, it appears that processing which is necessary to produce some of the spatial properties of the vertical VCR takes place outside the vestibular nuclei. Recording from pontomedullary reticulospinal neurons receiving vestibular input and projecting to different levels of the spinal cord reveals that almost no cells receive only vertical canal input, while approximately half receive otolith input. As is the case for vestibulocollic neurons, these reticulospinal neurons also lack the properties required to produce all of the VCR's spatial properties. Two conclusions are that in response to stimuli in vertical planes pontomedullary reticulospinal fibers are best suited to contribute to otolith reflexes, and that spatial properties of the VCR depend in part on convergence of inputs within the neck itself. PMID- 8234749 TI - Stance and balance following bilateral labyrinthectomy. AB - Although vestibular input codes head acceleration, it is not clear whether or not this signal is critical for triggering the initial postural response to a perturbation of stance, and for determining the appropriate direction of response. These experiments were designed to examine the contribution of vestibular inputs to the control of balance in the freely standing cat. Four cats were trained to stand quietly on a moveable force platform. The animal's stance was unexpected perturbed by applying a linear ramp-and-hold translation to the support surface in each of eight different directions in the horizontal plane. The characteristics of quiet stance and the response to the perturbations were quantified in terms of the 3-D ground reaction forces under each paw and the EMG activity in selected muscles. The animals were bilaterally labyrinthectomized, and their responses compared before and after lesion. The cats were able to stand stably on the platform within 2-3 days of the lesion. During quiet stance, there was no change in the distribution of vertical forces under the paws and no increase in sway area. Horizontal plane forces, which were normally outwardly directed on the diagonals, became more laterally directed and transiently larger in amplitude. The level of tonic EMG activity increased in some extensors and flexors, and decreased in others, compared to control. The responses to platform translation were characterized by normal spatial and temporal patterns and latencies of EMG activity. Furthermore, all cats continued to use the force constraint strategy that is characteristic of the intact animal (Macpherson, 1988a). The only clear deficit in performance was a transient hypermetria, characterized by an over-response to the translation. Although the cats over responded, they were still able to maintain their balance successfully. The moderate changes in quiet stance and in response to perturbation gradually returned to control values over 8-10 days following the lesion. These results suggest that vestibular information is not necessary for triggering appropriate postural responses evoked by support surface translations, nor for selecting the direction of response. PMID- 8234750 TI - Vestibular control of skeletal geometry in the guinea pig: a problem of good trim? AB - Motor control of different segments of the body with multiple degrees of freedom appears to be coordinated by utilizing preferred axes of motor activity. This hypothesis may also be applied to vestibular control of posture. To explore this question we studied the anatomical relationship between the head and the cervical vertebral column by taking radiographs of the head-neck region in unrestrained alert guinea pigs. We determined that biomechanical constraints contribute to the stereotypical skeletal geometry observed in the resting animal and to a functional segmentation of the head-neck movement apparatus. Subsequent lesion studies of vestibular end organs with quantification of the resulting postural syndromes suggest that the functional segmentation of the cervical vertebral column corresponds to a functional partitioning of vestibular afferents. Our findings also indicate that the sensorimotor transformation mechanisms necessary to convert a given head velocity signal into the appropriate neck motor frame are already embedded in the networks provided by second-order vestibular neurons. Good trim of postural control will be the end result of an appropriate internal representation of the objective vertical. PMID- 8234751 TI - Geometrical approach to neural net control of movements and posture. AB - In one approach to modeling brain function, sensorimotor integration is described as geometrical mapping among coordinates of non-orthogonal frames that are intrinsic to the system; in such a case sensors represent (covariant) afferents and motor effectors represent (contravariant) motor efferents. The neuronal networks that perform such a function are viewed as general tensor transformations among different expressions and metric tensors determining the geometry of neural functional spaces. Although the non-orthogonality of a coordinate system does not impose a specific geometry on the space, this "Tensor Network Theory of brain function" allows for the possibility that the geometry is non-Euclidean. It is suggested that investigation of the non-Euclidean nature of the geometry is the key to understanding brain function and to interpreting neuronal network function. This paper outlines three contemporary applications of such a theoretical modeling approach. The first is the analysis and interpretation of multi-electrode recordings. The internal geometries of neural networks controlling external behavior of the skeletomuscle system is experimentally determinable using such multi-unit recordings. The second application of this geometrical approach to brain theory is modeling the control of posture and movement. A preliminary simulation study has been conducted with the aim of understanding the control of balance in a standing human. The model appears to unify postural control strategies that have previously been considered to be independent of each other. Third, this paper emphasizes the importance of the geometrical approach for the design and fabrication of neurocomputers that could be used in functional neuromuscular stimulation (FNS) for replacing lost motor control. PMID- 8234752 TI - Quantitative comparison of electrically and acoustically evoked auditory perception: implications for the location of perceptual mechanisms. AB - Electrical stimulation of the human auditory system produces different patterns of spatial and temporal neural activity than those that occur in the normal, acoustically stimulated system. Quantitative comparison of psychophysical performance measured with acoustic and electrical stimulation may allow us to infer the physiological locus of perceptual mechanisms. In this paper we compare psychophysical data on temporal resolution from normal-hearing listeners, cochlear implant listeners, and patients electrically stimulated on the cochlear nucleus. Measures of gap detection, forward masking, and modulation detection will be compared. These comparisons demonstrate that temporal processing is relatively similar across these three groups once the obvious differences in dynamic range are taken into consideration. In addition, preliminary results with speech processors indicate that implant patients can utilize all temporal information in speech. Thus, implant patients have relatively normal temporal resolution and can integrate temporal cues normally for the recognition of complex acoustic patterns such as speech. These results imply that the central auditory systems of implant patients are able to fully utilize the non-natural patterns of temporal neural information produced by electrical stimulation. The differences in the microstructure of the neural pattern (phase locking, stochastic independence of fibers, spatial distribution of activity, etc.) between electrical and acoustic stimulation are apparently not necessary for temporal processing. Thus, the physiological locus of temporal processing mechanisms must be more central in the auditory system than the cochlea and cochlear nucleus. PMID- 8234753 TI - Pattern recognition and masking in cochlear implant patients. AB - Studies of the temporal course of masking using pulsatile electrical stimulation provide a sensitive new technique for the investigation of central pattern recognition. The masked threshold for a single-pulse probe was studied for several different maskers as a function of the time between the probe and the start of the masker. These experiments showed the gradual development of a temporal pattern in the masked thresholds as the number of pulses in the masker was increased. For a 210 msec masker with pulses at 10 msec intervals, both backward and forward masking thresholds showed a well-defined peak at times 10 msec before and after the masker. Probe pulses presented at these times were probably perceived to be part of the masker pattern and therefore were not easily identified as probe pulses. This conclusion was confirmed by using a masker with pulses at 20 msec intervals. Only backward masking was tested, and the results showed a peak approximately 20 msec before the start of the masker, fitting in with the temporal pattern of the masker. PMID- 8234754 TI - A digital speech processor and various speech encoding strategies for cochlear implants. AB - We distinguish two main categories in the speech decoding strategies: those based on feature extraction and those based on time information of the filtered incoming signal. In the first group only those electrode pairs corresponding to the localisation of the four or more, maximum spectral peaks or the formants of the speech signal are stimulated. The second category is essentially based on the timing information that is included in the filter outputs of the filtered speech signal. The Laura implant has the capability to be programmed to the different decodings-strategies. To understand the potential possibilities the main features of the implant are discussed. PMID- 8234755 TI - New hardware for analog and combined analog and pulsatile sound-encoding strategies. AB - Development for cochlear implants of primarily analog design focuses in two directions. The first direction is miniaturization. A behind-the-ear (BTE) speech processor has been developed which can replace the body-worn processor for approximately 90% of the users of a MED-EL cochlear implant and works with two 1.4 V hearing aid batteries for between seven and twelve days. Consonant, vowel and sentence testing and patient questioning revealed that the BTE speech processor demonstrates a significant improvement in speech understanding compared to the body-worn processor, and that the patients' device acceptance is superior for the BTE processor. The result of the second direction for our cochlear implant development is the multichannel cochlear implant, CAP, with combined analog and pulsatile stimulation. It aims at complementing the information from the broad-band analog signal by adding spectral information, that is, tonotopic information. This device is capable of simultaneously stimulating one electrode channel with a broad-band analog signal and one of eight electrode channels with a pulsatile signal. The system can also be used for purely analog or for purely pulsatile stimulation. Preliminary results with the first recipient of a CAP cochlear implant system demonstrate that the device works as expected. PMID- 8234756 TI - Speech encoding strategies for multielectrode cochlear implants: a digital signal processor approach. AB - The following processing strategies have been implemented on an experimental laboratory system of a cochlear implant digital speech processor (CIDSP) for the Nucleus 22-channel cochlear prosthesis. The first approach (PES, Pitch Excited Sampler) is based on the maximum peak channel vocoder concept whereby the time varying spectral energy of a number of frequency bands is transformed into electrical stimulation parameters for up to 22 electrodes. The pulse rate at any electrode is controlled by the voice pitch of the input speech signal. The second approach (CIS, Continuous Interleaved Sampler) uses a stimulation pulse rate which is independent of the input signal. The algorithm continuously scans all specified frequency bands (typically between four and 22) and samples their energy levels. As only one electrode can be stimulated at any instance of time, the maximally achievable rate of stimulation is limited by the required stimulus pulse widths (determined individually for each subject) and some additional constraints and parameters. A number of variations of the CIS approach have, therefore, been implemented which either maximize the number of quasi simultaneous stimulation channels or the pulse rate on a reduced number of electrodes. Evaluation experiments with five experienced cochlear implant users showed significantly better performance in consonant identification tests with the new processing strategies than with the subjects' own wearable speech processors; improvements in vowel identification tasks were rarely observed. Modifications of the basic PES- and CIS strategies resulted in large variations of identification scores. Information transmission analysis of confusion matrices revealed a rather complex pattern across conditions and speech features. Optimization and fine-tuning of processing parameters for these coding strategies will require more data both from speech identification and discrimination evaluations and from psychophysical experiments. PMID- 8234757 TI - Performance of the avian inner ear. AB - In spite of morphological similarities, the avian inner ear has apparently developed mechanisms of sound transduction that differ from the mammalian solution. This paper is a compilation of the present knowledge. PMID- 8234758 TI - New processing strategies for multichannel cochlear prostheses. AB - Various strategies for representing speech information with multichannel cochlear prostheses were compared in tests with implant patients. The strategies included the compressed analog (CA) approach of a standard clinical device, and alternative interleaved pulses (IP) and continuous interleaved sampling (CIS) strategies. CA and IP strategies had been compared in previous studies with a wide range of subjects. The present studies compared all three types in tests with one subject and CA and CIS strategies in tests with six additional subjects. Subjects for the present studies were selected for their excellent performance with the clinical CA processor, and the tests included closed-set identification of consonants and open-set recognition of words and sentences. For every test, every subject obtained his or her highest score, or repeated a score of 100% correct, using a CIS strategy. In the comparisons of all three approaches, IP processor scores were between those obtained with CA and CIS processors. The results are discussed in terms of their implications for processor design. PMID- 8234759 TI - Synergies and strategies underlying normal and vestibulary deficient control of balance: implication for neuroprosthetic control. AB - Future developments of neuroprosthetic control will probably permit locomotion and posture to be maintained without the aid of crutches and will therefore require some form of balance control. Three fundamental questions will arise. First, the question of the location of imbalance-sensing transducers must be assessed. Secondly, the synergy, which is the relative amplitude and timing of muscle activity, and/or the strategy of joint torques required to re-establish a stable posture for different types of balance disturbances must be addressed. Thirdly, the control laws that map either trunk muscle activity or imbalance sensing transducer outputs into multi-joint postural control of standing by paraplegic individuals must be generated. The most appropriate means of gathering the relevant information applicable to neuroprosthetic control systems is through the detailed analysis of normal and non-normal human models. In order to gain such detailed insights into normal balance control and its dependence on head angular and linear accelerations, the synergy and strategy of balance corrections in normal subjects or patients with vestibular deficits were investigated for two types of support surface perturbation, a dorsiflexion rotation (ROT) and a rearward translation (TRANS). These experimentally induced perturbations to upright stance were adjusted to cause equal amplitudes of ankle dorsiflexion, thus providing additional information about the role of lower leg proprioception on balance control. Synergies defined on the basis of peak cross-correlations of each recorded muscle's EMG to that of the largest muscle response were significantly different for TRANS and ROT. Translation synergies consisted of a sequential coactivation at several levels (soleus and abdominals some 30 msec before hamstrings, and trapezius some 15 msec before paraspinals), whereas the sequential activation of paraspinals and tibialis anterior dominated the balance synergy to ROT. Likewise, response strategies, defined using cross-correlations of joint torques, differed. That for TRANS was organised as a multi-link strategy with neck torques leading those of all other joints by 40 msec or more; hip joint lead ankle torques by 30 msec. That for ROT was organised around hip and ankle torques without a major correlation to neck torques. Vestibulary deficient subjects developed weaker synergies with respect to subjects with normal balance systems under eyes-open conditions and there was no clear synergy with eyes closed. Consequently, hip torques were delayed some 180 msec with respect to ankle torques, and correlations to neck torques were completely out of phase under eyes-closed conditions. Fundamental changes in TRANS synergies and strategies also occurred in vestibulary deficient subjects for eyes-open and eyes closed conditions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234760 TI - Human standing posture: multi-joint movement strategies based on biomechanical constraints. AB - We developed a theoretical framework for studying coordination strategies in standing posture. The framework consists of a musculoskeletal model of the human lower extremity in the sagittal plane and a technique to visualize, geometrically, how constraints internal and external to the body affect movement. The set of all feasible accelerations (i.e., the "feasible acceleration set" or FAS) that muscles can induce at positions near upright were calculated. We found that musculoskeletal mechanics dictate that independent control of joints is relatively difficult to achieve. When muscle activations are constrained so the knees stay straight, to approximate the typical postural response to perturbation, the corresponding subset of the feasible acceleration set greatly favors a combination of ankle and hip movement in the ratio 1:3 (called the "hip strategy"). Independent control of these two joints remains difficult to achieve. When near the boundary of instability, the orientation and shape of this subset show that the movement strategy necessary to maintain stability, without taking a step, is quite restricted. Hypothesizing that regulation of center-of-mass position is crucial to maintaining balance, we examined the feasible set of center-of-mass accelerations. When the knees must be kept straight, the acceleration of the center of mass is severely limited vertically, but not horizontally. We also found that the "ankle strategy", involving rotation about the ankles only, requires more muscle activation than the "hip strategy" for a given amount of horizontal acceleration. Our model therefore predicts that the hip strategy is most effective at controlling the center of mass with minimal muscle activation ("neural effort"). PMID- 8234761 TI - An integrated EMG/biomechanical model of upper body balance and posture during human gait. AB - Full scale biomechanical and EMG analyses of the balance during human gait are required to understand the neural control of locomotion. The purpose of this paper was to develop an inverted pendulum model of upper body balance in both the plane of progression and the frontal plane, and a medical/lateral balance model of the total body. EMG evidence was also recorded to reinforce the conclusions from the moment of force analyses. The kinematics and kinetics for up to ten natural walking trials on each of four subjects and EMG records from walking trials on eleven subjects were investigated. The results support the following conclusions. (1) The hip extensors/flexors have an over-powering role in maintaining dynamic balance of the head, arms and trunk (HAT) in the plane of progression. Because of the lack of suitable neurological and biomechanical delays between the small head acceleration, presumably exciting vestibular afferents, and the hip moment patterns, the vestibular system appears not to be involved as a feedback sensor in the balance control during gait. (2) In the frontal plane, the hip abductors are dominant in countering the large medial lateral (M/L) imbalance of HAT during single support but are assisted by the medial acceleration of the hip joint. (3) the total body M/L balance is achieved by the M/L placement of the foot with some opposition and some assistance by the M/L acceleration of the subtalar joint. The subtalar invertors/evertors play an insignificant role during single stance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234762 TI - Control of standing and gait using electrical stimulation: influence of muscle model complexity on control strategy. AB - Three approaches to controlling standing and gait in paraplegics through functional electrical stimulation are presented. The approaches differ in their requirements for modeling the muscle actuator. The first approach describes a detailed muscle model and presents methods for rapid experimental parameterization of the model. The second requires a less detailed model and knowledge of model error bounds to design advanced, non-linear controllers which guarantee stability. The third approach needs no muscle model since it controls limb trajectories through combining stimulation with an orthosis containing controllable friction brakes at the joints. Future clinical systems may use one or a combination of these approaches to restore useful function. PMID- 8234763 TI - FES gait restoration and balance control in spinal cord-injured patients. AB - The status of gait restoration in spinal cord-injured patients by means of FES is reviewed and the main aspects are discussed. This introduction highlights the issues of balance control, stimulation sequence synthesis, and control of enhanced gait modes containing unbalancing. The use of statically unstable dynamic weight-transfer phases is important for enhanced gait modes. To show how this phase can be employed the mode of static balance currently used for FES assisted four-point gait in paraplegic patients is discussed, and how this mode of gait can be converted to a semi-dynamic gait mode is described. The possibilities and consequences of such an approach are briefly discussed. PMID- 8234764 TI - Finite state model of locomotion for functional electrical stimulation systems. AB - A finite state model of locomotion was developed to simplify a controller design for motor activities of handicapped humans. This paper presents a model developed for real time control of locomotion with functional electrical stimulation (FES) assistive systems. Hierarchical control of locomotion was adopted with three levels: voluntary, coordination and actuator level. This paper deals only with coordination level of control. In our previous studies we demonstrated that a skill-based expert system can be used for coordination level of control in multi joint FES systems. Basic elements in this skill-based expert system are production rules. Production rules have the form of If-Then conditional expressions. A technique of automatic determination of these conditional expressions is presented in this paper. This technique for automatic synthesis of production rules uses fuzzy logic and artificial neural networks (ANN). The special class of fuzzy logic elements used in this research is called preferential neurons. The preferential neurons were used to estimate the relevance of each of the sensory inputs to the recognition of patterns defined as finite states. The combination of preferential neurons forms a preferential neural network. The preferential neural network belongs to a class of ANNs. The preferential neural network determined the set of finite states convenient for a skill-based expert system for different modalities of locomotion. PMID- 8234765 TI - Fatigue during functional neuromuscular stimulation. AB - Discontinuous activation of muscle compartments is used to postpone the occurrence of fatigue during both normal activation and artificial stimulation. Periods of force development are interrupted by passive periods during which the muscle can recover. Since it is not known how fatigue parameters depend on intermittent stimulation, we compared fatigue generated by continuous electrical stimulation with fatigue resulting from intermittent stimulation schemes. T5-T6 paraplegics participated in the experiments. Continuous stimulation generated a torque time course that can be described by a rising and a falling exponential time constant. The falling time constant ranged from 100 to 200 sec in all four patients and did not depend on the amplitude of the stimulation pulses. The torque developed at times greater than 250 sec was asymptotically constant and proportional to the maximal torque developed by each patient. Intermittent stimulation appears to postpone fatigue markedly. The torque time course developed in each on/off cycle again was a double exponential. In intermittent stimulation schemes average torque determines the muscle's performance. Therefore, average torque versus time was calculated from the intermittent stimulation data. These relations also follow a double exponential, accurately providing confident estimates of fatigue parameters. Fatigue caused average muscle torque to decline to a constant level which, for each patient, was uncorrelated with the duty cycle of the stimulation pattern. Between patients these levels varied from 18.3 +/- 7.1 to 9.9 +/- 4.3 (% of maximal torque +/- standard error). These findings result in a model that could be of use in controllers for functional neuromuscular stimulation (FNS) that take into account non-stationarity caused by fatigue. PMID- 8234766 TI - Mechanical demodulation of hydrodynamic stimuli performed by the lateral line organ. AB - Tonic displacements of the fish lateral line cupula were observed during stimulation of the organ with amplitude-modulated water motion. The modulation frequency was fixed at 2.4 Hz and the carrier frequency was varied from 25 to 500 Hz. The time waveforms of the cupular displacement at carrier frequencies below 280 Hz and above 470 Hz were essentially amplitude-modulated waves. Between 350 Hz and 410 Hz the magnitude at the modulation frequency increased sharply and the predominant shape of the displacement waveform changed to that of the modulating frequency. The mechanism for extraction of the modulation component may play a key role in the decoding of sensory information. PMID- 8234767 TI - Amplitude fluctuations of spontaneous otoacoustic emissions caused by internal and externally applied noise sources. AB - The simplest description for the generator of a spontaneous otoacoustic emission (SOAE) is that of a single (isolated) limit-cycle oscillator. Evidence is given that this description is too simple. And it is concluded that study of systems of coupled oscillators is needed to obtain more insight in the processes that generate SOAEs. Part of this conclusion is based on the study of properties of coupled electronic Van der Pol oscillators. PMID- 8234768 TI - Exploration of cochlear function by otoacoustic emissions: relationship to pure tone audiometry. AB - The amplitudes, growth functions and detection thresholds of evoked otoacoustic emissions (EOE) were measured in 44 normally hearing subjects and 138 patients with two categories of cochlear dysfunctions: (a) acoustic trauma; and (b) presbycusis. Separate sets of experiments were also performed: (a) detection of stimulus frequency emissions; and (b) click EOE. EOE properties were studied around 2 kHz, 1 kHz and 0.75 kHz (+/- 0.1 kHz). A partial correlation and multivariate analysis was carried out to investigate the relationships between EOE properties and puretone auditory thresholds (from 0.25 to 8 kHz, half-octave steps). For each experiment and each frequency, only one highly significant correlation was found, linearly relating the n kHz EOE threshold with the hearing threshold at 2n kHz: there was a shift of about one octave between EOE amplitudes and audiometric data. This means that EOE thresholds give no direct information about the local cochlear state. A simplified model has been implemented, which assumes that EOE thresholds and amplitudes are proportional to the total number of residual active sites along the organ of Corti, i.e., to the total length of active basilar membrane towards the base of the cochlea. It is shown that this model accounts for the results revealed by the statistical analysis and closely fits the experimental data. PMID- 8234769 TI - Efferent synapse mechanisms in chick hair cells. AB - Electrical stimulation of the efferent fiber to the hair cell is known to suppress peripheral auditory function. In isolated hair cells from the chick, acetylcholine (ACh) generated membrane hyperpolarization when a puff was applied at concentrations from 1 microM to 5 mM. Ionophoretic application of ACh generated a membrane hyperpolarization with a rapid time course. This is due to the increase of K conductance and is accompanied by the rise of the [Ca]i. PMID- 8234770 TI - Distortion-product otoacoustic emissions in normal and impaired ears: insight into generation processes. AB - Otoacoustic emissions can be used to study cochlear function in an objective, non invasive and rapid manner. These and other desirable features of emissions have inspired a significant amount of investigation into the practicalities of utilizing evoked emissions as clinical tests of hearing. Variables which affect the measurement of emissions can be sorted into two major categories consisting of factors affecting either emission generation or expression. The present report consolidates and summarizes recent findings of a series of experiments in our laboratory which address both the generation and expression of transiently evoked and distortion-product otoacoustic emissions. Because these two emission types have the greatest promise of becoming clinically useful, a complete understanding of the factors responsible for their measured properties is particularly important. PMID- 8234771 TI - A comparison of transiently evoked and distortion-product otoacoustic emissions in humans. AB - The measurement of transiently evoked otoacoustic emissions (TEOAEs) can identify a hearing loss exceeding 25-30 dB HL with high sensitivity. However, further quantification of the hearing loss is not possible, and the frequency specificity of TEOAEs has been questioned. Distortion-product otoacoustic emission (DPOAE) measurements are being developed for clinical use in the hope that they will be more frequency-specific than are TEOAEs. We have compared TEOAEs and DPOAEs in both normally hearing and hearing-impaired subjects with the purpose of learning more about the frequency specificity of these two types of emissions. In a first investigation, toneburst-evoked OAEs were compared to DPOAEs stimulated at 1, 2 and 4 kHz in ten ears without spontaneous otoacoustic emissions of ten normally hearing subjects. Input/output (I/O) functions of DPOAEs at frequency regions of 1 and 2 kHz were characterized by roll-overs and irregularities that were not present in either DPOAE I/O functions at 4 kHz or in TEOAE I/O functions at 1, 2 and 4 kHz. Mean slopes of the I/O functions increased with increasing frequency for DPOAEs and decreased for TEOAEs. In a second investigation, click-evoked OAEs and DPOAEs (stimulated by pure tones in the frequency range of 0.75-6 kHz) were measured in 42 ears of 21 normally hearing subjects and 128 ears of 64 subjects with varying degrees of sensorineural hearing loss. Results from both investigations revealed that the amplitude ratio between DPOAEs and TEOAEs changed systematically with frequency. DPOAE amplitudes became larger with increasing frequency and TEOAE amplitudes became smaller.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234772 TI - Nonsustained ventricular tachycardia and other high-risk predictors following myocardial infarction: implications for prophylactic automatic implantable cardioverter-defibrillator use. PMID- 8234773 TI - Risk identification for sudden cardiac death--implications for implantable cardioverter-defibrillator use. PMID- 8234774 TI - Cost-effectiveness of the implantable cardioverter-defibrillator. PMID- 8234775 TI - Prevention of sudden death in patients with coronary artery disease: the Multicenter Unsustained Tachycardia Trial (MUSTT). AB - This trial will significantly advance our understanding of the prognostic and therapeutic usefulness of electrophysiologic studies in patients with coronary artery disease. Several features of this trial are worth emphasizing. First, the protocol for performing programmed stimulation and serial drug testing is designed to mirror those currently in use by many practicing electrophysiologists. While practice patterns vary, the procedures used in the trial reflect what is considered "usual and standard" practice. Second, because half of the patients with inducible sustained ventricular tachycardia will be given no antiarrhythmic therapy, we will be able to ascertain the true risk of sudden death in this patient population without the influence of these agents. Third, this trial will assess the usefulness of a method of guiding antiarrhythmic therapy (electrophysiologic testing) to reduce mortality in this high-risk population. It will not evaluate the efficacy of a specific type of antiarrhythmic therapy. PMID- 8234776 TI - Future developments in implantable cardioverter defibrillators: the optimal device. AB - Despite recent therapeutic advances, SCD remains the leading cause of mortality in industralized nations. The most frequent cause of SCD is ventricular tachyarrhythmias in the setting of advanced structural heart disease due to chronic coronary heart disease or idiopathic dilated cardiomyopathy. Although high-risk groups can be prospectively identified, attempts at primary prevention have been largely unsuccessful. Effective treatment strategies for SCD survivors include antiarrhythmic drug therapy guided by programmed stimulation, endocardial resection, and ICDs. Device therapy has proven extremely effective in preventing recurrent sudden death from ventricular tachyarrhythmias. Widespread application of ICD therapy, perhaps even to include members of high-risk populations that have not experienced cardiac arrest, will depend on many factors including the demonstration that device therapy improves total mortality, not just arrhythmia related mortality, reduction in cost, and improvements in the devices themselves. Some of the important characteristics of the optimal ICD of the future are nonthoracotomy lead placement; subpectoral generator placement; multiprogrammable, tiered therapy; improved diagnostic specificity, whether based on electrogram or hemodynamic-sensing algorithms; improved integration of brady- and tachy-sensing systems; and enhanced electrogram storage capability with trans telephonic retrieval of electrogram recordings. The creation of this ideal ICD will obviously require continued technological advances; however, given the tremendous improvements realized over the first three generations of ICD systems, optimism for the future seems warranted. PMID- 8234777 TI - Electrocardiographic changes associated with acute cerebrovascular disease: a clinical review. AB - Patients with acute vascular disorders of the CNS demonstrate an abundance of both rhythm and morphologic changes in their ECG. Of these a few will demonstrate myocardial dysfunction and or damage. The value of the ECG in evaluating and predicting which patients will have myocardial dysfunction or damage is questionable. One would assume the echocardiogram would be of more help than the ECG in identifying patients with myocardial damage; however, little data are available. The reason for the poor correlation between ECG findings and clinical correlates has not been explained to date, but it is possible to postulate a theory. There are two mechanisms that might mediate ECG changes in these patients, ie, autonomic neural stimulation from the hypothalamus or elevated circulating catecholamines. Hypothalamic stimulation may cause ECG changes without associated myocardial damage whereas elevated catecholamines may result in myocardial damage. This might explain why so many patients have ECG changes and very few have demonstrable myocardial damage in general, or ischemic damage in particular. That cardiac antiischemic therapy does not change mortality may relate to the fact that treatment has been directed towards patients with ECG changes, which in turn do not correlate with myocardial damage. Better patient selection for such therapy might rest upon demonstration of wall motion abnormalities on echocardiogram. The weakness of this strategy is that many patients with stroke have preexisting coronary disease and wall motion abnormalities and thus echo findings may only document remote infarction rather than acute ischemia. PMID- 8234778 TI - Early intravenous beta-blocker combined with thrombolytic therapy for acute myocardial infarction: the Thrombolysis in Myocardial Infarction (TIMI-2) Trial. PMID- 8234779 TI - Body composition in elderly persons: a critical review of needs and methods. AB - Significant changes in body composition that have important health related effects are believed to occur in the elderly. Knowledge of these changes is important for diagnoses, prognoses, and treatment of health problems. Many health problems in the elderly could be prevented or alleviated by nutritional modulation, but better understanding of the nature, extent, and underlying physiology of body compositional changes is needed for such interventions to be successful. There are currently few data for body composition in the elderly, especially for those greater than 75 y in age, partly because conventional methods of assessing body composition are difficult to apply for technical and conceptual reasons. As a result, little is known regarding the relationships of body composition to nutritional, functional or health status in non-hospitalized, free-living elderly persons. Knowledge of the "natural history" of body compositional changes and their relationships to other nutritional and health factors could lead to new insights on prevention and treatment, the reduction of morbidity and extension of the quality of life of older persons. PMID- 8234780 TI - Diabetes mellitus. What have we learned from animals? AB - Progress in our understanding of the pathophysiology of diabetes mellitus has been made possible because of the availability of animal analogs of the various human diseases. Diabetes mellitus can be mild, moderate or severe depending on the genetic error that is responsible for the disease. Present estimates of errors that result in diabetes range from 20 to 100. Because similar errors have been found in spontaneously diabetic animals scientists have been able to identify the sequence of metabolic events and subsequent tissue change in many of these phenotypes. Studies of the efficacy of various drugs, diets and lifestyle choices on disease development and management thus were made possible. PMID- 8234781 TI - Adenoviral DNA integration and changes in DNA methylation patterns: a different view of insertional mutagenesis. PMID- 8234782 TI - Regulation of Bacillus subtilis gene expression during the transition from exponential growth to stationary phase. PMID- 8234783 TI - Genomic organization of T and W, a new family of double-stranded RNAs from Saccharomyces cerevisiae. PMID- 8234784 TI - Mechanism of action and regulation of protein synthesis initiation factor 4E: effects on mRNA discrimination, cellular growth rate, and oncogenesis. PMID- 8234785 TI - Enzymology of homologous recombination in Saccharomyces cerevisiae. PMID- 8234786 TI - Posttranscriptional control of the lysogenic pathway in bacteriophage lambda. PMID- 8234787 TI - Global regulation of mitochondrial biogenesis in Saccharomyces cerevisiae. PMID- 8234788 TI - DNA polymerase II, the epsilon polymerase of Saccharomyces cerevisiae. PMID- 8234789 TI - [Front line of life science. Discussion]. PMID- 8234790 TI - [What we can learn from the history of rubiscology]. PMID- 8234791 TI - [A turning point of the world]. PMID- 8234792 TI - [New developments of life science in connection with information science--future of bio-informatics]. PMID- 8234793 TI - [Circumstances before the appearance of protein structure in three dimension]. PMID- 8234794 TI - [Stick to the question of life sciences]. PMID- 8234795 TI - [Creation of paradigm and international evaluation of scientific contributions]. PMID- 8234796 TI - [Turning points on my way of biochemistry]. PMID- 8234798 TI - [Biochemistry with a simple mind]. PMID- 8234797 TI - [Signal transduction and cellular responses]. PMID- 8234799 TI - [Mammalian embryology and embryonic engineering: a new connection from basic biology to society]. PMID- 8234800 TI - [Cancer and biochemistry: recollection of personal 30 years' experience]. PMID- 8234801 TI - [Study of protein synthesis in animal cells]. PMID- 8234802 TI - [The future of molecular biology]. PMID- 8234803 TI - [Attractiveness of microorganisms]. PMID- 8234804 TI - [The substrate specificity of cytochrome P450]. PMID- 8234805 TI - [The continuity of life and mutations]. PMID- 8234806 TI - [Snake neurotoxins and their corresponding genes]. PMID- 8234807 TI - [From hiochic acid to tunicamycin]. PMID- 8234808 TI - [Morphology in molecular cellular biology]. PMID- 8234809 TI - [Study of DNA replication in search of universality and diversity]. PMID- 8234810 TI - [NMR structural analyses of proteins: scope and limitations. Discussion]. PMID- 8234811 TI - [Intracellular signaling pathway: function of GTP-binding proteins]. PMID- 8234812 TI - [Development of human genome study and the future of molecular biology]. PMID- 8234813 TI - [On cancer research--personal thoughts]. PMID- 8234814 TI - [Capillary electrophoresis]. PMID- 8234815 TI - [Interfacial electrokinetic phenomena--for better understanding of capillary electrophoresis]. PMID- 8234816 TI - [Capillary electrophoresis of nucleic acids]. PMID- 8234817 TI - [Capillary electrophoresis of proteins]. PMID- 8234818 TI - [Structure and molecular dynamics of smooth muscle myosin examined by using a zero-length cross-linker]. PMID- 8234819 TI - [Regulations of cell division cycles and MPF]. PMID- 8234820 TI - [Mitotic cyclins and MPF]. PMID- 8234821 TI - [Regulation of mammalian cell cycle progression by cdc 2 kinase and cdk]. PMID- 8234822 TI - [Paroxysmal nocturnal hemoglobinuria--molecular mechanism of abnormal hemolysis]. PMID- 8234823 TI - [Mammalian sperm-egg interaction--gamete maturation and potential role of oligosaccharides in fertilization]. PMID- 8234824 TI - [Hydration and conformational changes in polynucleic acids]. PMID- 8234825 TI - [Control of mRNA stability in plants--cis-acting elements which affect mRNA stability]. PMID- 8234826 TI - [cdc25, wee1, mik1]. PMID- 8234827 TI - [Microfilament system in cell cycle]. PMID- 8234828 TI - An inhibitor of thromboxane production attenuates tumor necrosis factor release by activated human alveolar macrophages. AB - Tumor necrosis factor alpha (TNF alpha) and thromboxane A2 (TXA2) are major products of the activated alveolar macrophage and serve as key mediators of lung injury. In order to determine if the synthesis of TXA2 and the release of TNF alpha are associated, the production of these inflammatory agents by the human alveolar macrophage (AM), as a result of activation by lipopolysaccharide (LPS), was assessed in the absence and presence of the thromboxane synthase inhibitors UK 38,485 (Dazmegrel) and OKY 046. UK 38,485 and OKY 046 inhibited both LPS stimulated TXA2 production and TNF alpha release in a dose-dependent manner. Prostaglandin E2 (PGE2) production was not increased by UK 38,485 or OKY 046. Neither LPS nor UK 38,485 had any effect on LTB4 production by AM. Neither UK 38,485 or OKY 046 had any effect on LPS-stimulated interleukin-1 beta release. However, the TXA2 mimetic, U46619, did not stimulate TNF alpha release by AM either in the absence or presence of UK 38,485. These findings suggest that 1) UK 38,485 and OKY 046 are inhibitors of both TXA2 production and TNF alpha release by activated human AM, 2) UK 38,485 probably does not exert its inhibitory action on TNF alpha release through effects on eicosanoid production and 3) the possibility that TNF alpha- and TXA2-induced lung injury may be subject to amelioration by imidazole-based compounds should be further evaluated. PMID- 8234829 TI - Characterization of alveolar macrophage eicosanoid production in a non-human primate model of mineral dust exposure. AB - The relative activation of eicosanoid production which results from the exposure of the alveolar macrophage (AM) to mineral dusts is thought to be a key factor in the pathophysiology of occupational lung disease. We compared in vitro basal and silica-stimulated production of prostaglandin E2 (PGE2) and thromboxane A2 (TXA2) by AM from normal humans and non-human primates (Macaca nemestrina). In addition, we instilled mineral dusts directly into one lung of the non-human primate and evaluated AM eicosanoid production at two week intervals following dust instillation. Unstimulated AM from humans produce more PGE2 and TXA2 than do AM from M. nemestrina. However, in vitro exposure of AM from both species to silica dust produced a qualitatively similar increase in TXA2 production accompanied by no change in PGE2 production. Sequential analysis of AM eicosanoid production following a single bolus exposure to bituminous or anthracite coal dusts, titanium dioxide (TiO2) dust or crystalline silica showed marked variability among individual non-human primates in qualitative and quantitative aspects of dust-induced eicosanoid production. However, the rank order of potency of the different dusts (silica > anthracite > bituminous) correlated with epidemiological evidence relating the type of dust mined to the incidence of pneumoconiosis. These studies suggest that the non-human primate may serve as a model for the study of both the role of eicosanoids in the etiology of dust induced occupational lung disease and the biochemical basis for individual variability in the response of lung cells to mineral dust exposure. PMID- 8234830 TI - Stereospecific actions of misoprostol on rat colonic electrolyte transport. AB - Misoprostol (Miso) produces a mild, transient diarrhea in some patients, which is believed to be partly due to intraluminal fluid accumulation. To better understand this diarrheagenic action, we compared the effects of Miso, its 4 stereoisomers (11R16R, 11R16S, 11S16S, 11S16R), misoprostol free acid (Miso-FA), and 16,16-dimethyl PGE2 (dmPGE2) on rat colonic electrolyte transport in vitro. Increases in short-circuit current (Isc) were measured (after serosal addition) in segments of mucosa stripped of muscularis and mounted in Ussing chambers. The rank order of apparent potencies, in terms of threshold, were (microM): 11R, 16S (1.2) approximately dmPGE2 (1.0) > Miso-FA (10.0) approximately Miso >> 11R, 16R; 11S, 16R; 11S, 16S (all inactive at 100 microM). The response to dmPGE2 and Miso was attenuated in the presence of the Na+/K+/Cl- co-transport inhibitor bumetanide (100 microM). Pretreatment with atropine (0.1 microM) did not affect the Isc response to Miso, Miso-FA, or dmPGE2. Tetrodotoxin partially attenuated (39 +/- 9% inhibition) the response to Miso-FA, but did not affect Miso or dmPGE2. In conclusion, Miso increases Cl- secretion across rat colonic mucosa through a direct action on epithelial cells. The activity resides in the 11R,16S isomer, thus implying a stereospecific interaction at PGE receptors. The effect of Miso to stimulate epithelial Cl- secretion might contribute to its diarrheagenic action in vivo. PMID- 8234831 TI - Myocardial salvage efficacy of a thromboxane receptor antagonist, SQ, 30,741, in relation to inhibition ex vivo of platelet function in the ferret. AB - The myocardial salvage efficacy of a thromboxane A2/prostaglandin endoperoxide (TP) receptor antagonist has not been previously determined in a ferret model of ischemia and reperfusion. Assessments of the reproducibility of infarct size resulting from a 90 min period of occlusion followed by 5 hr of reperfusion of the left anterior descending coronary artery in saline-treated control ferrets revealed a consistent mean level of tissue damage representing 23.1 +/- 1.4% of the left ventricle. In subsequent studies, ferrets were given the thromboxane receptor antagonist SQ 30,741 (1 mg/kg bolus and 1 mg/kg/hr infusion, i.v.) or vehicle. At this dose, SQ 30,741 significantly reduced infarct size from that measured in control ferrets by 44%. Concurrently, the drug produced a 97% inhibition of platelet TP receptors as measured by inhibition of the ex vivo platelet shape change response to U-46,619. Drug administration was not associated with measurable alterations in mean blood pressure, heart rate or the rate-pressure-product. The importance of this finding to clinical utility and the mechanism of the observed cardioprotective action, however, remain unclear. These data indicate that the ferret represents a useful model for the assessment of the myocardial salvage efficacy of TP receptor antagonists and are consistent with attenuation of ischemic myocardial damage by doses of these agents which produce > 96% TP receptor blockade. PMID- 8234832 TI - Effect of prostaglandin E2 on rate of decidualization in rats. AB - Based on morphology, it has been suggested that prostaglandin E2 (PGE2) accelerates the process of endometrial stromal cell differentiation to decidual cells in the rat. The present study investigated this possibility, using changes in uterine weight and in endometrial alkaline phosphatase (ALP) activity as indicators of decidualization. Rats were ovariectomized and treated with one of two steroid protocols; the first was identical to that previously used for the study of morphology, the second a modified protocol which results in greater uterine sensitization for decidualization, producing larger amounts of decidual tissue, thereby making it easier to detect differences between treatments. On the day of uterine sensitization, rats within each treatment protocol were given a unilateral intrauterine infusion of phosphate-buffered saline (PBS) or PGE2 plus indomethacin, and killed 24, 48 or 72 h later. The time-courses for the increases in uterine weight and ALP activity in uterine horns infused with PBS or PGE2 plus indomethacin differed between steroid protocols, but within a protocol were statistically indistinguishable. The results do not support the hypothesis that PGE2 accelerates the process of decidualization but do provide additional support for the notion that PGE2 is a physiological rather than pharmacological mediator of decidualization. PMID- 8234833 TI - Modulation of human myometrial PGE2 receptor by GTP characterization of receptor subtype. AB - We studied PGE2 specific binding sites in human myometrial microsomes prepared from uterine specimens obtained by hysterectomy (women between 38 and 55 years of age). Competition experiments showed that the potency order for various prostaglandins (PGs) was: PGE2 > or = PGE1 >> PGF2 alpha > Iloprost > or = Carbacyclin >> ZK 110841 (PDG2 analogue). These relative affinities indicated that the receptor was of the EP type. In kinetic experiments GTP, GppNHp and GTP gamma S increased the rate of PGE2 binding (steady state was reached more rapidly in the presence of nucleotides) but maximal specific binding was not significantly different. Complete dissociation could not be obtained, even in the presence of GTP. Only 50% of maximal binding was readily dissociable. The dissociation rate was 4.56.10(-4) sec-1 (half time of about 660 sec) and in the presence of GTP analogues it was slightly increased (k-1 = 7.16 10(-4) sec-1, half time 420 sec.). Scatchard analysis of saturation curves showed an increase in ligand receptor affinity in the presence of GTP or nucleotide analogues: the Kd shifted from 9.66 +/- 2.8.10(-9) M to 4.96 +/- 1.25.10(-9) M, but the number of binding sites did not change significantly (310 +/- 37 to 350 +/- 17 fmol/mgP). The effect of GTP was observed at a concentration of 5.10(-4)M. GppNHp and GTP gamma S were effective at 1.10(-5) M. Pretreatment of myometrial membranes with pertussis or cholera toxins had no effect on PGE2 binding to membrane sites. Our conclusion is that GTP induced conversion of a population of low affinity sites into a population of higher affinity sites. This effect of guanine nucleotides was described in adipocytes and kidney medulla. Competition studies with PGE2 analogues (sulprostone, 17-phenyl-omega-trinor PGE2, M&B 28,767, misoprostol, butaprost) showed that this receptor mediates a contractile response and is probably an EP3 subtype. PMID- 8234834 TI - Constriction of canine coronary arteries by platelet activating factor after brief ischemia. AB - Platelet activating factor (PAF) has been suggested as a mediator of coronary spasm and acute myocardial ischemia. However, PAF can have either vasodilator or vasoconstrictor activities according to various reports. Because of the importance of endothelium in regulating vascular tone, we hypothesized that changes in endothelial function could modulate some of the observed differences in the activities of PAF. To test this hypothesis, PAF was infused directly into the left anterior descending coronary artery (LAD) of dogs at a rate of 0.3 microgram/min before and after 20 min of LAD ligation followed by reperfusion. Coronary blood flow (CBF) was measured continuously via a Doppler flow probe. Likewise, responses to the endothelium dependent vasodilator, acetylcholine (ACh) were measured before and after the LAD ligation. Before ligation, PAF produced a vasodilatory response in the LAD, resulting in 28.4 +/- 11.0% increase in CBF and a 21.5 +/- 5.8% decrease in coronary vascular resistance (CVR). However, after ligation and subsequent reperfusion, 0.3 microgram/min PAF produced vasoconstriction, resulting in a 10.2 +/- 8.7% decrease in CBF (p < 0.01 compared to pre-infusion change in (CBF), and a 27.8 +/- 23.2% increase in CVR (p = 0.05) compared to pre-infusion change in CVR). The vasodilator response to ACh was markedly blunted by ischemia. These results suggest that the coronary vascular response to PAF may depend upon the functional integrity of the endothelium, with endothelial damage resulting in constrictor responses to PAF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234835 TI - Effect of prostaglandin F2 alpha on uterine or ovarian secretion of prostaglandins E and F2 alpha in vivo in 90-100 day hysterectomized, intact or ovariectomized pregnant ewes. AB - Vehicle or 8 or 16 mg of PGF2 alpha per 58 kg body weight was given intramuscularly to intact, hysterectomized or ovariectomized 90-100 day pregnant ewes in three separate experiments. Both doses of PGF2 alpha increased PGF2 alpha in ovarian venous plasma compared with controls at 72 hr post treatment in intact (P < or = 0.05) but did not in hysterectomized (P > or = 0.05) 90-100 day pregnant ewes. Concentrations of PGE in ovarian venous blood of intact ewes did not differ (P > or = 0.05) between treatment groups and were equivalent to concentrations of PGE determined in uterine venous plasma. PGE was decreased in ovarian venous plasma by PGF2 alpha in hysterectomized ewes (P < or = 0.07). PGE in uterine venous plasma averaged 6 ng/ml over the 72-hr treatment period in intact and ovariectomized 90-100 day pregnant ewes and was 12 fold greater (P < or = 0.05) than PGF2 alpha which averaged 500 pg/ml in uterine venous plasma. Both PGF2 alpha and PGE increased (P < or = 0.05) by 64 hr in uterine venous plasma of the 8 mg PGF2 alpha-treated intact pregnant ewes. A significant quadratic increase (P < or = 0.05) was observed for PGF2 alpha and PGE in the vehicle and both PGF2 alpha treatment groups of intact ewes at the end of the 72 hr sampling period. It is concluded that the uterus and ovaries secrete significant quantities of PGF but little PGF2 alpha during midgestation. In addition, PGF2 alpha increased uterine secretion of PGE in vivo. PGE may be a placental stimulator of ovine placental secretion of progesterone or PGE may protect placental steroidogenesis from actions of PGF2 alpha. PMID- 8234836 TI - [National School of Hygiene]. PMID- 8234837 TI - [Program for elimination and eradication of poliomyelitis caused by wild polioviruses in Poland]. AB - Since 1988 the programme coordinated by the WHO of elimination and eradication poliomyelitis by the year 2000 is performed. In this article the following topics are presented: 1) the criteria of elimination and eradication of poliomyelitis caused by wild polioviruses; 2) the epidemiological situation in the world, European Region and Poland; 3) the role and activities of epidemiologists, clinicians and virologists in the programme. PMID- 8234838 TI - [Factors constituting a threat to humans of rabies in Poland]. AB - Today, there are two steps of human rabies impendence: a. exposures to rabid or rabies suspected animals b. human deaths due to rabies. Red foxes, dangerous rabies reservoir are responsible for the main tendency and spread of rabies in Poland (over 2000 cases in 1992). This has a direct bearing on the treatment of exposed persons and on rabies among unvaccinated domestic animals. The number of persons requesting post exposure rabies treatment is also increasing--6500 vaccinees in 1992. During last eight years no case of human death was registered in Poland. Mass parenteral canine vaccination campaign and effective control methods of rabies in dogs in Poland are the mostly successful and significant elements separating wild animal rabies from human society. Any inaccuracy on this field would be the risk of transmission of wild rabies to human surrounding. PMID- 8234839 TI - [An epidemiologic assessment of viral hepatitis type A morbidity in the Gdansk province in the years 1989-1990]. AB - The considerable increase in viral hepatitis type A morbidity was observed in 1989-1990 in the area of Gdansk province. That increase was clearly focusing, it concerned only the chosen towns and communities. In these areas and also in regions where the increase in morbidity did not take place, the special researches have been carried out among children and adults which determined the frequency of passed HAV infection. It has been tried to investigate the ways and to describe the reasons of epidemic enlargement. It was proved that in adults group, the difference of the passed infection HAV frequency was not significant statistically comparing to the towns and villages dwellers, either for the regions where epidemic took place or without it. Comparison of the examined children revealed statistically the essential differences between the towns and villages-dwellers and significantly higher anti-HAV frequency among the children from the areas with morbidity increase. For explanation of the epidemic enlargement ways, the fast spreading of infection was emphasised on the areas situated at Wierzyca river and all its tributary streams. PMID- 8234840 TI - [Routes of transmission of hepatitis B in Piotrokow and Sieradz provinces in 1986 1987]. AB - Epidemiological investigations of 1883 cases of acute viral hepatitis type B reported in two voivodships (Piotrkow, Sieradz) has been analysed. In 1104 cases (58%) parenteral procedures for therapeutical and diagnostic purposes have been performed during 6 months before the onset of disease. The incidence of cases after medical procedures was the highest in children aged 0-4 years (67%) and adult persons aged over 50 years (68%). Home contacts with HBV carriers and/or recoverers gave only 342 cases (18.2%). Professional infections occurred in 117 medical workers (6.2%). At that time both voivodship had high hepatitis B morbidity ranging 91.7/100,000 in Piotrkow voivodship in 1986 over two times higher than in the whole country (38.9/100,000). PMID- 8234841 TI - [Course of acute hepatitis B in children]. AB - This study presents the course of acute hepatitis B in 77 children. It has been recorded that the course of HBV infections in children is most often mild, and in children younger than 2 years of age it is usually inapparent. The most frequent markers of the infection are antigens HBs and HBe. ALAT in all children normalized 2 months at the longest, after the hospitalization had ended. The most considerable danger of carrier-state exists in children not older than 2 years of age; an increased danger of becoming a carrier exists during the time of pubescence. The main cause of HBV infections is hospital stay. There emerges a necessity of routine marking the HBs antigen in hospitalized children. PMID- 8234842 TI - [Hepatitis B virus infection in children]. AB - Retrospective analysis of 256 medical records of children with confirmed diagnosis of hepatitis B virus infection was done. The study focused on significance of patient's age and sex in the course of the disease, on circumstance accompanying the HBV infection and on course of infection, especially a chronic HBV infection. It has been found that factors increasing the risk of infection include: former hospitalizations, surgical procedures, malfunctions of an immune system and familial contacts. Hepatitis occurred more frequently in infants and young children and in this age it more often converted into a chronic form. The course of hepatitis infection depended on dynamics of the disease process. Seroconversion occurred more often after acute phase of the disease and asymptomatic course of infection most often resulted in chronic form. Treatment with immuno-potent (TFX, isoprinosine) had no effect on the course of hepatitis B infection. It should be recommended to extend prophylaxis to children of the risk group and to introduce routine test for HBs antigen in each inpatient child. PMID- 8234843 TI - [Evaluation of seroconversion after voluntary vaccination against hepatitis B in students at the University of Lodz]. AB - We have evaluated the results of the first action of voluntary vaccination against hepatitis B in the students of Medical University of Lodz using Energix-B vaccine (20 micrograms) in schedule 0, 1, 6 months. Few students applied for the vaccination: only 48.4% of persons planned for vaccination completed the schedule. Seroconversion rate (anti-HBs > 10u/l) in this group was 93.9%. Three doses of vaccine applied inconsistently with schedule resulted in the immunological response in 3 out of 4 students observed. Two doses of vaccine resulted in the effective response in 13 out of 16 students observed. One doses of vaccine proved to be ineffective. The blood tests performed prior to vaccination showed the presence of the HBV markers in 9.6% of students (HBsAg in 2.1% of persons and anti-HBs in 5.3/of persons). PMID- 8234844 TI - [Awareness of AIDS in selected population groups from the province of Lublin]. AB - The poll carried out among selected community groups in the Lublin Voivodship has shown that the problem of AIDS is looked upon multifariously depending on age. The paper is an attempt to compare opinions of the 18-25 age group (118 people) with those of the 40-60 age group (100 people). The poll, which consisted of 8 questions, has indicated that the knowledge of epidemiology and preventive measures in reference to AIDS is incomplete, the younger group revealing greater awareness of the problem. It has also been determined that the need for information on the HIV virus is high. It has been agreed that television and radio programmes constitute the best methods to convey information on AIDS; the press comes second followed by various publications and open lectures. The poll has also shown that the young tend to accept HIV carriers more than the 40-60 age group. PMID- 8234845 TI - [Influenza and other viral respiratory infections in recruits during the epidemiologic season of 1991-1992 in Poland]. AB - Viral Respiratory Infections in recruits have been presented as appearing in the epidemical season of 1991-1992. Three singular parainfluenza Type 1, Type 3 and RS virus-infections have been found. No influenza viruses were isolated. PMID- 8234846 TI - [Evaluation of seroconversion after vaccination against influenza during the epidemic season 1991-1992 in Poland]. AB - Vaccination was performed by influenza virus vaccine trivalent type A and B. We vaccinated about 75,000 persons in Poland in different groups of age. HI and NI tests were used for testing antibody level for A/Taiwan/1/86 (H1N1), A/Beijing/353/89 (H3N2) and B/Panama/45/90. The rise of antibody level was statistically significant. PMID- 8234847 TI - [Poisoning after eating sausage in observations from I Clinic of Infectious Diseases of the Slask Academy of Medicine in 1985-1992]. AB - On the basis of our own material and the literature review the analysis of 10 cases of botulism is presented. A mild course of the disease was observed in 40%, a medium-severe course in 30% and a severe course also in 30% of patients. Fatal courses were not recorded. All patients were discharged from the Clinic in the good state of health. Etiopathogenesis, symptomatology and treatment of botulism is discussed. Attention is paid to new diagnostic method. PMID- 8234848 TI - [Characteristics of Salmonella strains isolated from different environments in the years 1982-1993]. AB - The examination of Salmonella strains isolated from hospital patients and out patients was performed. The results of sensitivity tests of strains isolated in Poland, and from patients of "Third World countries" were compared. In both groups were the strains caused nosocomial infections, which were resistant to Ampicillin and Chloramphenicol. Eight of the Polish strains of S. Typhimurium were cultured from autopsy specimens which were obtained from the newborn ward. S. Typhimurium isolated from specimens of the patients of the children's, ward of Derna Hospital was the cause of 162 cases on nosocomial infections over the period of four months. The ill children were between the ages of the two days old (newborn child in incubator) and eleven years old. It is important to note that in one case S. Enteritidis was isolated from the blood of a hospital patient in critical condition. A much higher percentage (85%) of the strains isolated from out-patients in Poland showed resistance to Tetracycline in comparison with the strains cultured from out-patients in countries of the Third World (7%). The resistance to Tetracycline may be connected with the addition of this antibiotic to food for farm animals in Poland, and can indicate the circulation of animal strains in the human population. In this research we used the latest classification of Salmonella genus. PMID- 8234849 TI - [Toxicity of aldehydes used as disinfectants]. AB - The paper presents a review of current scientific literature on the toxicology of aldehydes used as cold disinfectants. A particular attention was paid towards the mutagenic, teratogenic and cancerogenic potential as well as occupational hazard and risk for patients exposed to aldehydes. PMID- 8234850 TI - [Evaluation of disinfection procedures conducted in hospitals in Poland from the aspect of hospital infection control]. AB - The physical and chemical procedures of disinfection presently employed in hospitals are described. It is emphasized that not all technical and organizational appliances, currently in use, satisfy the requirements. Propositions aimed at improving this situation are presented. PMID- 8234851 TI - [A case of infectious mononucleosis syndrome caused by cytomegalovirus]. AB - A case of 26-years old man with symptoms of infectious mononucleosis syndrome is presented. In the course of the disease: high temperature, weakness, loss of appetite, sore throat, myalgia, hepatomegaly, splenomegaly and some laboratory changes (leucocytosis with presence of atypical lymphocytes, elevated aminotransferase activity) have been observed. Serological tests have shoved: slighthly positive PBD-test in the first examination (second-negative) and the presence of IgM antibodies against CMV in a high titer with four-time decrease of the titer during the course of the disease. Because of the incomplete symptom complex for infectious mononucleosis caused by EBV we have put the diagnosis of cytomegaly coursed as a infectious mononucleosis syndrome. PMID- 8234852 TI - [Impressions of training at the infectious diseases hospital unit in Lyon]. PMID- 8234853 TI - [Social differentiation of increases in low birth weight in Poland--biological consequence of recession?]. AB - The reasons of the increase in the LBW rate in Poland from 8.1% in 1985 to 8.4% in 1990 was investigated using the individual records from the birth certificates. There was differential increase of the low birth weight among the social groups. The highest increase was observed among the less educated mothers, especially in big cities. The LBW among the newborns delivered by mothers who had finished their education at the primary school level changed from 10.6% (in big cities from 14.7%) in 1985 to 12.5% (in big cities to 16.2%) in 1990. PMID- 8234854 TI - [Effect of psychological family crises on manifestations of Alzheimer's disease in people of working age]. AB - Retrospective data concerning the presence of psychological family crises during one year preceding the onset of the Alzheimer's have been analysed in a group of 73 patients. A diagnostic of the disease (revealing of the symptoms) defined the "onset" of the illness in this study. The results demonstrated that the psychological family crises are an important element of the etiopathogenesis of the Alzheimer's disease. Their better understanding might be therefore an important factor in preventing the illness. Since psychological crises are commonplace in the widely understood human population, a prospective study investigating their coincidence with the onset of the Alzheimer's disease might bring further interesting results and is being suggested by the author. PMID- 8234855 TI - MR imaging of pleural and peritoneal effusion. AB - The purpose of this study was to assess the efficacy of MR imaging in predicting the quality of ascites and pleural effusion. MR examinations of 20 patients with different benign or malignant diseases accompanied with ascites or pleural effusion were retrospectively studied. Results were compared with histopathological findings. On T1- and T2-weighted images, the MR intensity of the effusion depended mainly on the concentration of protein, whereas on gradient echo images, it depended mainly on the concentration of blood. MR examination was useful in non-invasively differentiating exudative or hemorrhagic effusion from serous effusion. Knowledge or inference of the quality of effusion can direct MR interpretation and may improve diagnostic accuracy. PMID- 8234856 TI - Correlation of sequential MR imaging of the injured spinal cord with prognosis. AB - Forty-nine patients with acute spinal cord injuries were studied sequentially with MR imaging by using 0.5 Tesla superconductive units, and sequential MR changes were correlated with the prognosis of the patients. MR images were obtained within one week of the injury and then every two to six months when possible. The Frankel classification of neurologic function was correlated with MR findings. The most frequently observed types of signal intensity patterns on MR imaging were type 0 (isointensity on both T1- and T2-weighted images) and type I (isointensity on T1- and hyperintensity on T2-weighted images). In subsequent subacute and chronic stages, type II (hypointensity on T1 and hyperintensity on T2) was most frequently observed. The evolution of type 0 was to types I and II, whereas type I usually turned into type II or remained as type I. Type III (hyperintensity on T1 and hyper-, iso- or hypointensity on T2 images) patients were few in number. There was a good correlation between MR imaging patterns and neurologic recovery for initial and subsequent MR patterns, in that type 0 showed good recovery, whereas types I and II revealed good improvement or no recovery. In addition, the extent of the high signal intensity area on initial as well as on subsequent T2-weighted images was proportionally correlated to neurologic recovery. The degree of cord compression was also important for predicting recovery of neurologic function. Findings of MR imaging of acutely injured spinal cord suggested the prognosis of spinal cord injury, especially when sequential studies were obtained. PMID- 8234857 TI - Local control of prostate cancer with transurethral intracavitary radiation therapy. AB - As of January 1986, 13 patients with adenocarcinoma of the prostate had been treated in our clinic by remote after-loading transurethral high dose rate radiotherapy using a 60Co source. Of these patients, four were at stage B2, three at stage C, three at stage D1, and three at stage D2. The mean total dose of transurethral radiotherapy was 35.2 Gy to the most distant prostatic capsule from the source. Three patients with stage D1 disease and one patient with stage C disease received additional external radiation with a total dose of 20 Gy to the prostate and 40 Gy to the pelvis. Local tumor response proved rapid and satisfactory as verified by rectal examination, ultrasonography, and needle biopsy. Serious complications such as proctitis, cystitis, incontinence, and urethral stricture were not evident during the average follow-up term of 34.9 months. PMID- 8234858 TI - Disseminated necrotizing leukoencephalopathy following irradiation and methotrexate therapy for central nervous system infiltration of leukemia and lymphoma. AB - The authors report four adult patients with disseminated necrotizing leukoencephalopathy (DNL) following therapy for central nervous system (CNS) involvements of leukemia and lymphoma. Five to fourteen months after starting therapy with 30.6-62.5 Gy of whole brain irradiation and 120-500 mg of intrathecal methotrexate (MTX), DNL developed. Brain CT scans demonstrated a characteristic symmetrical white matter low density area. Furthermore, the brain CT scans disclosed tumorous lesions with contrast enhancement in three cases. In two of the three cases autopsy proved the tumorous lesions to be necrotic foci but not leukemic tumors. Post-mortem neuropathological studies of three patients disclosed characteristic features of DNL: multiple coagulative necrosis in the white matter with myelin degeneration, swollen axons, prominent calcification, and enlarged reactive astrocytes. Three of the four patients obviously received greater doses of whole brain irradiation and intrathecal MTX than patients who remained free from DNL after treatment with whole brain irradiation and/or MTX. Analysis of the etiological factors in this series underscores the prevailing danger of treatment for neoplastic CNS involvement with excessive doses of whole brain irradiation combined with intrathecal MTX. PMID- 8234859 TI - Chronological observation in early radiation myelopathy of the cervical spinal cord: gadolinium-enhanced MRI findings in two cases. AB - Gd-enhanced MR images of two patients with clinically and histopathologically diagnosed chronic progressive radiation myelitis (CPRM) were observed chronologically. One of them had had nasopharyngeal cancer and received radiotherapy at a dose of 100 Gy to the C1-2 level of the spinal cord. She developed CPRM 25 months after the termination of radiotherapy. The other had had malignant lymphoma originating from the tonsil and received chemoradiotherapy. The dose delivered to her cervical spinal cord was 40 Gy, and she developed CPRM 30 months later. Gd-enhanced MRI in the early phase revealed a small crescent shaped nidus with or without a very small central nonenhanced area in both cases. Enhancement was not great. At that time, patients noticed only the inability to perceive pain and temperature or paresthsia in the opposite side. In a few months, MRI revealed a much more strongly enhanced and larger nidus with enlargement of a central nonenhanced area accompanied by long segmental cord enlargement. The patients' neurological symptoms had also progressed, with hemiparesis developing, for example. PMID- 8234860 TI - Hypertensive encephalopathy: reversible CT and MR appearance. AB - The authors present a case of hypertensive encephalopathy in which CT and MR were performed. The brain stem and cerebellum were revealed as low density areas on CT and high intensity areas on T2-weighted MR images with mass effect. Slight abnormality was also noticed in the deep cerebral white matter. This distribution pattern is contrary to that described in previous reports in which cerebral white matter was mainly involved. The lesions showed definite improvement when the blood pressure was normalized. MR is more sensitive than CT, and is useful for early detection of the abnormality and follow-up study. PMID- 8234861 TI - Repositioning of a displaced catheter for percutaneous hepatic arterial infusion chemotherapy. AB - This is a report of a patient with hepatocellular carcinoma whose hepatic arterial catheter, which was introduced via a subclavian arterial branch, migrated into his right gastric artery. The catheter was successfully maneuvered to a suitable position without any further invasive procedure. The importance of preparatory embolization of the right gastric artery is underscored. PMID- 8234862 TI - Isolated Crohn's disease of the gastroduodenum: a case report. AB - We report a case of isolated Crohn's disease in the gastroduodenum. A 33-year-old man was hospitalized due to epigastralgia. Endoscopic and double-contrast radiographic studies revealed edematous gastroduodenal mucosa with a deep, undermining ulceration in the gastric antrum, which changed after a short interval into a typical "cobblestoned" appearance. A gastrectomy was performed due to antral narrowing, and Crohn's disease was histologically confirmed. Our objective was to reveal the change from a deep, undermining ulceration to a typical "cobblestoned" appearance using a double-contrast barium study. PMID- 8234863 TI - Hepatic granuloma with progressive calcification: CT appearance. AB - Calcification in the liver is uncommon but not rare in clinical practice, and may be caused by many pathological conditions. We report a patient with hepatic mass on abdominal sonogram and CT scan. The mass was shown as a hypoechoic lesion on abdominal sonogram and as a well-marginated hypodense mass on pre- and postcontrast CT. Inflammatory granuloma was diagnosed by pathological examination following percutaneous needle biopsy. A follow-up precontrast CT scan was performed 22 months after the initial examination, and the hepatic granuloma could be the cause of hepatic parenchymal calcification. PMID- 8234864 TI - Distribution of radiolabeled cis-diamminedichloroplatinum (II) on scintigraphic images in the tumor-bearing rabbit. AB - To evaluate the feasibility of using scintigraphic images to assess the distribution of cis-diamminedichloroplatinum (II) (CDDP), we obtained serial scintigraphic images using radiolabeled CDDP (Pt-195m CDDP), which had high specific activity (7.4 x 10(6) Bq [200 microCi]/mg-CDDP) in the rabbit VX-2 tumor system. Radioactivity at the heart decreased in a biexponential manner, with an initial half-time of 15.3 min and secondary phase of 4.5 days. Time-activity curves revealed that the tumor/heart ratio was greater than 1.0 (1.0-1.1) between 30 min and 4 hr after intravenous injection. The scintigraphic study clearly depicted the tumor. Pt-195m CDDP uptake by the rabbit tumor at 2 hr was 7.8% of the total injected dose. These data suggest that scintigraphic imaging using Pt 195m CDDP is a promising method for imaging tumors and evaluating their CDDP uptake. PMID- 8234865 TI - Beneficial liaisons: radiobiology meets cellular and molecular biology. AB - The scientific knowledge that is emerging in all fields of medicine is rapidly changing our understanding of the concepts in radiation oncology. In this review, some of the classic radiation biology theories and models are examined and newer 'models' are illustrated. The ability of radiation oncologists to remain current with the newer scientific findings is essential to the development of improved therapeutic strategies and, importantly, to the proper balance between investment in technology and biology. PMID- 8234866 TI - Radiation dose assessment in radioiodine therapy. Dose-response relationships in differentiated thyroid carcinoma using quantitative scanning and PET. AB - Dose-response charts have been constructed to determine the tumouricidal dose for differentiated thyroid carcinoma metastases and thus enable precise activities of radioiodine to be prescribed in order to maximise tumour kill and minimise morbidity. Tumour and normal residual thyroid absorbed doses from radioiodine-131 have been determined with increased precision using a dual-headed whole-body rectilinear scanner with special high-resolution low-sensitivity collimators. Improved accuracy in the estimation of functioning tumour mass has been achieved using positron emission tomography (PET) with a low-cost large area PET camera. Dose-response data have been obtained for 33 patients. Following near-total thyroidectomy and 3.0 GBq 131I, a mean absorbed dose of 410 Gy achieved complete ablation of thyroid remnants in 75% of patients. Patients who had persistent uptake in the thyroid region on subsequent radioiodine scanning had received a mean dose of only 83 Gy. Cumulative absorbed doses in excess of 100 Gy were found to eradicate cervical node metastases. Patients with bone metastases, who generally have a poor prognosis, were found to have received doses of the order of only 20 Gy to the tumour deposits. The dose-response data explain the spectrum of clinical responses to fixed activities of radioiodine. In future, they will enable precise prescription of radioiodine to achieve tumouricidal doses whilst avoiding the morbidity and expense of ineffective therapy. PMID- 8234867 TI - Black and white in accuracy assessment of megavoltage images: the medical decision is often grey. AB - Using different criteria for acceptance of the portal film taken at the first treatment session, a comparison was made of the relevance of the information obtained from such a single check for the subsequent irradiations. A total number of 234 verification films have been taken on 29 fields for 27 head and neck patients. Patients were immobilised with individual plastic masks fixed to the couch and treated on a 6-MV linac fitted with an automatic verification system. Field alignment was checked with a measurement in the anteroposterior (AP) and craniocaudal (CC) direction on each film. Referring to the simulated field, this group of patients was treated with excellent average precision (mean, -0.7 mm) and reasonable spread (s = 5 mm). The percentage of 'large' deviations (> or = 6 mm) occurring during the whole treatment course is proportional to the upper limit of deviation accepted in the first assessed field (for an upper limit < 6 mm): it goes progressively up from 5% (AP-CC direction) to 17% (AP) and 13% (CC) for accepted magnitudes of deviation going from 2 mm to 6 mm in the first film. As the reproducibility of the different treatment series (s = 2 mm) is independent of the upper level of error accepted on the first film, this means that errors are mainly systematic errors coming from the transfer of the simulation unit to the treatment unit. Precision in a series of set-ups is always expressed by a Gaussian curve.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234868 TI - A dosimetric quality audit of photon beams by the Belgian Hospital Physicist Association. AB - Since 1987, a dosimetric standardization process has been started by the Belgian Hospital Physicist Association (BHPA). As part of this project, on-site visits were performed by a voluntary team, by request of the local physicist. They included mechanical checks on treatment machines and simulators and a dosimetric intercomparison of photon beams, following the EORTC methodology. Until now 16 centres have participated. Dose measurements were performed in order to evaluate the uniformity in dosimetry after the adoption by the BHPA members of the Nederlandse Commissie voor Stralingsdosimetrie (NCS) code of practice for photon beams. Doses were measured with an ionization chamber in water under NCS reference conditions and under other conditions in 13 cobalt beams and 24 high energy X-ray beams from 4 to 25 MV. Under reference conditions, the mean ratio of measured to stated dose is 0.999 (+/- 0.010) and 1.006 (+/- 0.23) for cobalt and X-ray beams, respectively, with a difference between the extreme values (delta) of 0.042 for cobalt beams and 0.084 for X-ray beams. In other conditions, the spread is a little larger for all beams. The results are comparable, but with a smaller dispersion, with those of other national and international similar dose intercomparisons and show the importance of applying a common protocol. The results of mechanical checks show some large deviations in digital displays and indicate the need for a systematic mechanical and beam alignment quality assurance programme. PMID- 8234869 TI - Relative biological effectiveness of intermediate energy protons. Comparisons with 60Co gamma-radiation using two cell lines. AB - Range modulated proton beams are used for radiotherapy of malignant tumours at several accelerator laboratories with the aim of introducing proton therapy as a clinical hospital-based therapy modality. Due to the finite range and the sharpness of the dose gradients, the dose to well defined target volumes can be raised without excessive irradiation of non-target tissue. The prescribed proton doses are determined in part on the basis of the relative biological effectiveness (RBE) of the particular radiation quality. In this study, RBE values were determined for a proton beam with a maximal range of 33 mm, which corresponds to an energy of approximately 67 MeV. The range modulated depth-dose distribution, with a 20 mm extended Bragg peak, was mainly designed for high precision treatment of small targets such as uveal melanomas. The tested cell lines, LS-174T and V79-379A, were chosen because of their suitability for clonogenic assays. The cells were irradiated with single doses in the range 2-10 Gy at different depths in the extended peak region of the range modulated proton beam. RBE values were determined by comparing the doses needed to obtain the same reduction in colony formation (0.5, 0.1 and 0.01) as with the reference 60Co gamma source. The mean RBE value was 1.22 with a standard deviation of 0.08. The variations depended on both cell type and on the survival levels considered.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234870 TI - Effect of small doses per fraction in rat spinal cord: influence of initial vs. final top-up doses. AB - Previous data suggested that the linear-quadratic (LQ) model underestimated the sparing effect of small doses per fraction on the radiation tolerance of rat cervical spinal cord when 24 h was allowed between fractions for repair. In these experiments, animals had been given initial top-up doses consisting of 3 daily fractions of 9 Gy to represent 75% tolerance, followed by small fractionated doses in 1, 10, 20, 30 and 40 fractions given once daily. The end-point was forelimb paralysis secondary to white matter necrosis. To assess the possible perturbation of the initial top-up doses on the biological system, an experiment was performed with the small fractionated doses given initially, followed by the same top-up (final top-up) doses. The ED50s for 1, 10, 20, 30 and 40 fractions followed by final top-up doses were 9.5 +/- 0.3, 22.6 +/- 0.6, 32.4 +/- 0.6, 37.7 +/- 0.8 and 41.7 +/- 0.9 Gy, respectively; the corresponding ED50s obtained from the initial top-up experiment were 10.0 +/- 0.4, 20.7 +/- 0.5, 30.0 +/- 0.8, 37.0 +/- 0.8 and 39.9 +/- 0.7 Gy for 1, 10, 20, 30 and 40 fractions, respectively. Using the direct method of analysis and assuming complete repair between fractions, data from both experiments were not adequately described by the LQ model, which gave small alpha/beta values of 0.97 Gy for the initial top-up experiment and 1.23 Gy for the final top-up experiment, in contrast to an alpha/beta value of 2.41 Gy for the experiment with full course fractionation, fraction sizes down to 2 Gy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234871 TI - Enhancement of tumor cell killing in vitro by pre- and post-irradiation exposure to aclacinomycin A. AB - Aclacinomycin A (ACM), a potent inducer of leukemic cell differentiation, significantly enhances the radiosensitivity of a human colon tumor cell line (Clone A) when cultures are exposed to 15-nM concentrations for 3 days before irradiation. We now demonstrate that incubation with ACM after irradiation can also enhance Clone A cell killing. The maximum increase in cell killing, based on colony-forming ability, occurred when Clone A cells were exposed for 1 h to 5 microM ACM added 1 or 2 h after irradiation. The post-irradiation ACM protocol reduced the terminal slope (as reflected by D0) of the radiation cell survival curve with no change in the low-dose, shoulder region of the curve (Dq value). In contrast, for pre-irradiation treatment with ACM (15 nM, 3 days), the shoulder region of the curve was reduced with no change in the terminal slope. For pre- and post-irradiation ACM treatment the dose enhancement factors at 0.10 survival were 1.22 and 1.28, respectively. When ACM was given both before and after irradiation both the shoulder and terminal slope values decreased to produce a dose enhancement factor at a surviving fraction of 0.10 of 1.50. These data suggest that the enhanced cell killing produced by pre- and post-irradiation treatment with ACM is achieved through different mechanisms. PMID- 8234872 TI - Relationship between radiobiological hypoxia and direct estimates of tumour oxygenation in a mouse tumour model. AB - CDF1 mice were allowed to breathe either oxygen, carbogen or different concentrations of carbon monoxide gas. These treatments significantly altered the percentage of clonogenic hypoxic cells in a C3H mouse mammary carcinoma and these changes were found to correlate with direct measurements of tumour oxygenation obtained using an Eppendorf pO2 electrode. PMID- 8234873 TI - Tumor control probability for permanent implants in prostate. AB - The tumor control probabilities (TCPs) for 11 transperineal permanent implants of the prostate with 125I seeds have been computed, based on target-specific volume dose histogram data. Prostate dose-response of this modality is converted from that of external beam therapy based on biologically effective dose derived from the alpha beta model. The calculated TCPs for these transperineal prostate cases are compared with the results of 679 previous retropubic implants, providing interesting insight about these two techniques. Further refinement and validation of this model await long-term followup results of sufficient number of patients. PMID- 8234874 TI - Quality audit of megavoltage radiotherapy units: intercomparison of dose at a reference point using a mailed TL-dosimetry system. AB - A dosimetry intercomparison based on mailed TL-dosimeters and the well proven IAEA/EORTC phantom is described. Its aim is to identify discrepancies in dosimetry larger than +/- 3%. Dosimeters were mailed to all radiotherapy centres in Switzerland for irradiation with 2 Gy at a reference point in a water container, using photons and electrons. Thirty-six beams were monitored. The results show an agreement of within 2% for the majority of the beams monitored. Two electron beams were at 6% of the reference value. PMID- 8234875 TI - Analysis of dosimetric measurements in linac radiosurgery calibration. AB - The aim of this paper is to analyse the dosimetric parameters of a linear accelerator used in radiosurgery treatments. The influence of these parameters on the resulting dose distribution are basic for delivering the predefined dose to the vascular or oncological target volume. Several dosimetric methods have been used to define the output factors for small fields. The thimble and the Markus chambers, TLD and film dosimetry are investigated; all these dosimetric systems give reliable and almost similar results if used in the correct way. In the determination of tissue maximum ratio (TMR) the response curves obtained by ionometric and film dosimetry were investigated. For TMR determination the use of the Markus chamber and the correction factors to be applied as a result of the small dimension of the field were also investigated. PMID- 8234876 TI - Human errors in the calculation of monitor units in clinical radiotherapy practice. AB - Human mistakes are an important source of error in all steps of radiotherapy planning and their incidence should be investigated. As has been recognized by different authors and by the ICRU [4], the human error rate in the calculation of monitor units (MU) is relatively high. At our institute, we measured the human error rate in the calculation of MU by an independent check of the calculation. From September 1991 to June 1992 we identified and corrected 17 serious errors (deviation from the prescribed dose > or = 5%) over 890 controls (1.9%) (daily dose errors). We also found a serious global dose error rate (i.e. the errors induced on the total reference dose for the complete course of the treatment) of 1.3% (9/685) during the period November 1991-June 1992. These values suggest the importance of human errors in the calculation of MU and also confirm the validity of the independent check of MU calculation as one of the simplest ways of avoiding erroneous dose delivery by incorrect calculation of MU. PMID- 8234877 TI - Case #6. Smokeless tobacco keratosis. PMID- 8234878 TI - Keys to the profession. PMID- 8234879 TI - The write stuff. PMID- 8234880 TI - Nothing to fear but fear itself. PMID- 8234881 TI - More research needed to judge when to use chemical curettage. PMID- 8234882 TI - Suit up in suitable gear to guard against infection. PMID- 8234883 TI - Success, failure of implants depends on meticulous care. PMID- 8234884 TI - Build new bridges without tearing down the old ones. PMID- 8234885 TI - The unique stability of the marsupial sperm acrosomal membranes examined by unprotected freeze-thawing and treatment with the detergent Triton X-100. AB - In this study of the unique stability of the marsupial acrosome, experiments were carried out on the acrosomes of spermatozoa of the tammar wallaby (Macropus eugenii), common brushtail possum (Trichosurus vulpecula) and grey short-tailed opossum (Monodelphis domestica). Light microscopy showed that 4% of opossum and 15% of possum and wallaby spermatozoa lost their acrosomes after freeze-thawing. Electron microscopy revealed that freeze-thawing also induced changes in the acrosomal matrix of some acrosome intact spermatozoa. In both possum and wallaby, freeze-thawing increased the number of spermatozoa with vesiculation of the acrosomal matrix. Freeze-thawing disrupted the plasma membrane of spermatozoa but the acrosomal membranes remained intact. Immediately on addition of high concentrations of TX-100 (0.02% and 0.04%) there was significant loss of acrosomes and motility in possum and wallaby spermatozoa. Lower concentrations of TX-100 (< or = 0.01%) did not affect motility for up to 30 min in all three species, and there was no significant loss of acrosomes. Although loss of acrosomes did not occur under mild detergent treatment, 56% of wallaby and 70% of possum spermatozoa had altered acrosomes after 30 min in 0.01% TX-100. Electron microscopy revealed that acrosomes were undergoing a vesiculation process similar to that seen after freeze-thawing. Often the plasma membrane of detergent-treated spermatozoa was disrupted and had formed plasma membrane vesicles. However, the acrosomal membranes remained intact despite major changes to the acrosomal matrix. The study confirmed the remarkable stability of the marsupial acrosome and suggested that this is probably based in the acrosomal membranes. PMID- 8234886 TI - Tissue and endocrine responses to gestrinone and danazol in the treatment of endometriosis. AB - An open comparative study of gestrinone and danazol in the treatment of endometriosis demonstrated a similar decrease in the extent of active endometriotic deposits, assessed visually following therapy. Biochemical endocrinological changes during therapy with gestrinone showed that basal concentrations of gonadotrophin remained unchanged, oestradiol concentrations varied and the free fraction of testosterone increased. A similar range of side effects was reported in both treatment groups, consisting predominantly of 'androgenic' effects in 10 of 15 women treated with gestrinone and 5 of 12 treated with danazol. The results suggest that gestrinone is an effective alternative agent for the treatment of endometriosis. PMID- 8234887 TI - Catabolic utilization of glucose by the sheep conceptus between days 13 and 19 of pregnancy. AB - The production of carbon dioxide and lactate from glucose by sheep embryos and samples of extraembryonic membranes was measured during a 2.5 h incubation period. Both embryos and their membranes were active in the glycolytic and oxidative utilization of glucose and, in general, the utilization of glucose per unit weight fell as development progressed from Day 13 to Day 19 of pregnancy. Both oxidation of glucose and glycolysis by the extraembryonic tissues, expressed as activity per microgram dried tissue, fell progressively with development. The rate of decline in CO2 production was greater than the rate for glycolysis and, as a consequence, the contribution of glycolysis to the estimated energy yield from the catabolism of glucose rose with time. In the embryo, both glucose oxidation and glycolysis peaked on Day 15 with estimates of adenosine triphosphate (ATP) production from glucose per microgram dried tissue on this day being 50% above those on Day 13 and 100% above those on Day 17. In general, the estimated yields of ATP from glucose were similar for structures of the same developmental age except that, at Day 19, it was calculated that the rate of ATP production by embryos was double that by the extraembryonic membranes. In incubations using 5.56 mM glucose as sole exogenous energy source, glucose turnover by embryos and embryonic membranes tended to be higher in a bicarbonate buffered medium than in HEPES (4-(2-hydroxyethyl)-1-piperazincethane sulfonic acid) and phosphate-buffered media. As a result, the estimate of ATP yield plus the contribution of oxidative pathways to this yield were significantly higher in this medium than in the others. Glucose turnover by the embryo and its membranes in bicarbonate-buffered medium containing 0.56 mM glucose plus the alternate substrates, lactate and pyruvate, was severely depressed. Further experiments using samples of trophoblast and yolk sac indicated that both reduction in glucose concentration and the presence of the other substrates contributed to this suppression. Furthermore, an interaction between these factors was evident with the effects of alternative substrates being exaggerated when glucose concentration was low. PMID- 8234888 TI - Oxidative utilization of glucose, acetate and lactate by early preimplantation sheep, mouse and cattle embryos. AB - The production of radiolabelled CO2 from [U-14C]glucose, [1-14C]lactate, and [U 14C]acetate was used to study the oxidative metabolism of embryos recovered from sheep, mice and cattle. Sheep embryos showed an increasing capacity to oxidize glucose after the 4- to 8-cell stage and oxidative turnover of this substrate at the blastocyst stage was four times that at the early stages. Decarboxylation of carbon-1 of lactate followed a pattern similar to that seen for glucose oxidation, but acetate oxidation was low and did not follow the trends with development seen for the other substrates. Furthermore, estimates of incorporation of acetate into the macromolecules of sheep embryos were low compared with similar estimates for glucose and, unlike glucose, did not increase with development. Oxidation of all three substrates by mouse embryos increased with development but the rate of CO2 production from acetate was low compared with that from the other substrates. A combination of lactate or glucose with acetate had no influence on the utilization of acetate by mouse morulae/early blastocysts, nor did acetate influence utilization of the other substrates. Cattle morulae/early blastocysts also produced more CO2 from glucose and lactate than from acetate and the incorporation of carbon from acetate into the macromolecules of these embryos was less than from glucose. Overall, cattle embryos showed the greatest ability to metabolize acetate but were not as effective in using the other substrates as either sheep or mouse embryos at the same stage. PMID- 8234889 TI - Anti-sperm antibodies and semen profiles in re-anastomosed men. AB - A group of 29 re-anastomosed men were examined with respect to semen quality, anti-sperm antibody titres in serum and seminal plasma, presence of anti-sperm antibodies on sperm, and success rate in inducing pregnancy. Results indicated no association between pre-reversal serum anti-sperm antibody titres and post reversal semen quality, but a pregnancy induction rate of zero was associated with serum anti-sperm antibody titres greater than 160. It is recommended that men considering reversal, with anti-sperm antibody titres of this level, should receive counselling about the possibility of post-reversal infertility. PMID- 8234890 TI - Production and secretion of progesterone in vitro and presence of platelet activating factor (PAF) in early pregnancy of the marsupial, Macropus eugenii. AB - The corpus luteum (CL) of the tammar is suppressed by lactation. Removal of the sucking pouch young induces reactivation of the CL which produces a peak in plasma progesterone concentration on Day 5, 6 or 7; reactivation of the embryo after diapause precedes by one day this transient peak of progesterone. This study examines progesterone production and secretion in vitro by the CL and the production of platelet activating factor (PAF) by the endometrium during this stage of pregnancy (Days 0 and 3-8). Blood samples were collected twice daily to determine for each animal (n = 28) at autopsy the relationship of PAF production to the day of the progesterone peak. Significant changes in basal plasma progesterone concentrations were used to define four groups according to the time the animals were killed: Group A, Day 0; Group B, other animals showing no significant change; Group C, peripheral progesterone concentration increasing; Group D, peripheral progesterone concentration decreasing after the peak. At autopsy on successive days, blood samples were taken from the heart, caudal vein and uterine branch of the ovarian vein, and CL and endometria were prepared for explant culture. Progesterone contents of pre- and post-incubation luteal tissues and of medium were determined at 4, 12 and 24 h to estimate production and secretion rates. The values for all progesterone parameters from animals in Group C were significantly higher than in the other groups. It is concluded that the progesterone peak results from a change in rate of production and secretion of progesterone by the CL, one day before the peak in peripheral plasma, and that this change could provide the first signal to the uterus. Endometrium was incubated for 24 h and the medium assayed for PAF. High concentrations of PAF were detected in half the animals in Groups B and C, that is, before or at the time progesterone was increasing. This is the first measurement of PAF in any marsupial and the first demonstration of the release of PAF by the endometrium. The detection of PAF provides a candidate for an endometrial signal that is responsive to the rising progesterone concentration in the utero-ovarian vein draining the CL-bearing ovary and that may stimulate reactivation of the blastocyst. PMID- 8234891 TI - Ovarian function and its manipulation in the tammar wallaby, Macropus eugenii. AB - This study aimed to develop a superovulation protocol for the monovulatory tammar wallaby (Macropus eugenii), and examined the regulation of ovarian activity which leads to alternate ovulation in this marsupial. The most effective stimulatory treatment was 20 I.U. pregnant mare serum gonadotrophin (PMSG) given intramuscularly (i.m.) 20 days after the activation of an oestrous cycle by the removal of a sucking pouch young (RPY). Bromocriptine treatment was given at the time of RPY if the animal was in early seasonal quiescence. Mating had generally occurred when animals were examined 2 days after PMSG treatment on the morning of Day 22 RPY. Ovulation occurred only if the animal was treated on Day 22 or 23 (i.e. 2 or 3 days after PMSG) with gonadotrophin releasing hormone (GnRH) to induce a luteinizing hormone (LH) surge. Three 30-micrograms injections of GnRH (in 0.2 mL olive oil) were delivered as i.m. injections at 3-h intervals. Radioimmunoassay confirmed that the PMSG dose used did not elevate circulating steroid hormone concentrations beyond those found in normal cycles and that the GnRH protocol led to an LH surge of at least 6 h. Although multiple ovulation was achieved, the number of ovulations was low (2 or 3 per female). A major factor influencing the low ovulation rate was that generally only one ovary responded. Fertilized eggs and cleaving embryos were obtained. However, the fertility of induced ovulations has not yet been examined systematically. Laparoscopic examination through successive natural cycles confirmed that follicle growth and ovulation in the tammar wallaby alternates between the right and left ovary. Inhibition of follicle development in the corpus luteum (CL)-bearing ovary was also seen in females treated with the exogenous gonadotrophin (PMSG/GnRH) superovulation protocol. Follicle development was inhibited during the first half of the cycle in the non-CL-bearing ovary and during the entire cycle in the CL bearing ovary. This inhibition seemed to occur at the follicular level because exogenous gonadotrophin was unable to initiate a response during periods of inhibition and the response to gonadotrophin differed in the two ovaries. The number of follicles growing in the non-CL-bearing ovary in response to an exogenous gonadotrophin stimulus was inversely related to the weight of the growing CL for the first 19 days after RPY.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234892 TI - Supersensitivity to the stimulant action of noradrenaline on human myometrium near term. AB - Noradrenaline (10-50 nM) and tyramine (0.05-1 mM) enhanced contractile force elicited by field stimulation of strips of myometrium from non-pregnant and pregnant women. In higher concentrations, noradrenaline produced sustained contractions. The EC50 values for noradrenaline were 0.4 microM in tissues from pregnant women and 3.1 microM in tissues from non-pregnant women; maximum responses were greater in the former tissues. In addition, the effects of noradrenaline on myometrium from pregnant women were more marked on the inner layer than on the outer layer, antagonized by the alpha 1-adrenoceptor antagonist prazosin (0.1 and 1.0 microM), and unaffected by the inhibitor of neuronal uptake, nisoxetine (0.1 microM). Taken together, these observations confirm that supersensitivity to noradrenaline develops during pregnancy and is present near term. The supersensitivity to noradrenaline at term can be attributed only in part to a decrease in its removal by the sympathetic innervation, which declines towards term, because responses to tyramine were also enhanced in tissues from pregnant women. It is possible that gap junction formation may also contribute to this supersensitivity. PMID- 8234893 TI - Use of single or multiple injections of FSH in embryo collection programmes in goats. AB - The yield of embryos from goats treated with single injections of FSH combined with low doses of PMSG was compared with that from goats treated with conventional superovulatory regimens based on multiple doses of FSH. In one experiment, 101 female goats were assigned to 10 groups. One group received six injections of FSH-P (FSH-P control); a second group received eight injections of Ovagen (Ovagen control). The remaining eight groups conformed to a 2x2x2 factorial design in which goats received a single injection of a low or high dose of either FSH-P or Ovagen in combination with a low or high dose of PMSG. All goats were run with harnessed, entire bucks. Ovaries were examined by laparoscopy and embryos were recovered from goats that had ovulated and been mated. The mean number of normal embryos collected per goat was significantly greater in the FSH P control group in all except those groups treated with either low or high doses of PMSG combined with a low dose of Ovagen or a low dose of PMSG combined with a high dose of Ovagen. Most goats ovulated and ovulation rates were not significantly different from the mean of the FSH-P control, except for those treated with a low dose of FSH-P combined with either low or high doses of PMSG. Embryo recovery rates were significantly lower in groups treated with a high dose of PMSG combined with a high dose of either Ovagen or FSH-P, or with low doses of PMSG and FSH-P, than in the FSH-P control.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234894 TI - Fertility of superovulated ewes after intrauterine or oviducal insemination with low numbers of fresh or frozen-thawed spermatozoa. AB - Two experiments investigated the effects of number of spermatozoa inseminated and time and site of insemination of fresh and frozen-thawed semen on fertility in Merino ewes treated with intravaginal progestagen pessaries and a combination of pregnant mare serum gonadotrophin and follicle stimulating hormone. In Experiment 1, ewes were inseminated in the uterus or oviducts with totals of 10(4), 10(5), 10(6) or 10(7) fresh spermatozoa 44 or 68 h after pessary removal. Ova recovered 48 h later were classified as fertilized if they had cleaved. Proportion of fertile ewes (ewes with fertilized ova per ewes inseminated) and fertilization rate of ova (ova fertilized per ova recovered) were higher for inseminations 44 h (63% and 45%) than 68 h (38% and 19%) after pessary removal (P < 0.001). More ewes were fertile after oviducal (61%) than after intrauterine insemination (39%; P < 0.05), and with high (10(6) and 10(7)) than with low (10(4) and 10(5)) doses of spermatozoa for intrauterine (54% v. 24%; P < 0.05) but not for oviducal inseminations (63% v. 59%). Fertilization of ova was better after oviducal than after intrauterine inseminations (44% v. 22%; P < 0.001), and with high than with low doses of spermatozoa (45% v. 21%; P < 0.001). In Experiment 2, ewes were inseminated with 10(5), 5 x 10(5) or 10(6) total fresh or frozen-thawed spermatozoa in the uterus or oviducts, 44 h after pessary removal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234895 TI - Relationship between clustering of post-ovulatory and hierarchical follicles and their order of ovulation in the mature domestic hen (Gallus gallus domesticus). AB - Hierarchical (> 8 mm) and post-ovulatory follicles from the ovaries of laying hens at 45 and 86 weeks of age were assigned to clusters defined by the coalescence of their follicular stalks. They were also classified by size to give the succession in which ovulation had occurred, or would occur for the hierarchical follicles. The association between the order of ovulation and the observed clusters was compared with randomness and with dependence on the cluster in which the immediately preceding ovulation occurred. The order of ovulation was associated randomly with clusters, and was independent of the cluster containing the preceding ovulation. Many perceived clusters of successive follicles and post ovulatory follicles may be compatible with a random order of ovulation. These findings do not support theories of stimulation or inhibition of adjacent follicles, leading to eventual ovulation, by a dominant follicle. PMID- 8234896 TI - Effect of prepubertal gonadectomy and sex steroid treatment on the growth and lymphocyte populations of the rat thymus. AB - The age at which rats are most sensitive to prepubertal gonadectomy, in terms of thymic growth, was investigated. Gonadectomy enhanced thymic growth at each age; the greatest difference in thymic weight between gonadectomized and intact animals occurred in male rats gonadectomized at 5 weeks of age (64%) and in female rats gonadectomized at 4-5 weeks of age (43-46%). The effect of various synthetic sex steroids on growth and lymphocyte populations of the thymus in gonadectomized rats was examined. Diethylstilboestrol, 1 mg per animal, inhibited thymic growth by more than 35% in both males and females. Ethinyloestradiol, 40 micrograms per animal, inhibited thymic growth by 26% in males but by only 4% in females. Fluoxymesterone, 10 mg per animal, inhibited thymic growth by more than 46% in both sexes. Norgestrel, 12 micrograms per animal, had no effect on thymic growth. The synthetic steroids that significantly inhibited thymic growth decreased the intensity and altered the localization of staining for total T cells (antibody clone MRC OX 19), T helper cells and macrophages (W 3/5), T cytotoxic/suppressor cells (MRC OX 8) and B cells (MRC OX 12). PMID- 8234897 TI - Porcine granulosa cell conditioned media as autocrine regulator of progesterone secretion. AB - The ability of porcine granulosa cells to release a progesterone inhibiting substance(s) was examined in vitro. Granulosa cells (SGCs, MGCs and LGCs) were harvested from small, medium or large antral follicles respectively. The effect of granulosa cell conditioned media obtained from small follicles (SGCCM) was studied in the culture of LGCs by estimation of progesterone secretion; the conditioned media evoked the inhibition of progesterone secretion by the LGCs. SGCCM produced by various numbers of cultured granulosa cells showed a dose related inhibition of progesterone production. A maximum inhibitory effect was noted when a 5-fold concentration of SGCCM was used. The addition of SGCCM had no effect on the growth of the cultured cells. The factor(s) inhibiting progesterone secretion appeared to be a nonsteroidal substance of molecular mass greater than 10 kDa and was heat-stable and trypsin-sensitive. The data presented support the suggestion that the conditioned media generated by primary cultures of SGCs contain nonsteroidal regulators capable of inhibiting progesterone secretion by cultured LGCs; this inhibitory activity can play an important autocrine regulatory role in the process of follicular differentiation. PMID- 8234898 TI - Corticotropin-releasing hormone stimulates adenylyl cyclase activity in the retinas of different animal species. AB - In the present study we investigated the presence of corticotropin-releasing hormone (CRH)-stimulated adenylyl cyclase activity in the retinas of different animal species. CRH significantly stimulated adenylyl cyclase activity in homogenates of calf, pig, rabbit and guinea pig retinas. The stimulatory effects were concentration-dependent with half-maximal responses occurring at 20-30 nM CRH. The enzyme activities increased by 37-80% at the maximal concentration of CRH (1 microM). On the other hand, adenylyl cyclase activities of chicken and pigeon retinas were poorly stimulated by CRH. In calf, pig and rabbit retinas, the CRH effect was completely antagonized by the CRH receptor antagonist alpha helical CRH 9-41 and required the presence of GTP. The stimulatory response elicited by CRH was also found to be not additive with that produced by either vasoactive intestinal peptide or dopamine. These results provide evidence for the presence in retinas of different animal species of functional CRH receptors, an important criterion for the classification of CRH as a retinal neurotransmitter. PMID- 8234899 TI - Effects of central neuromedin B and related peptides on blood glucose. AB - Bombesin (Bn), a peptide of amphibian origin, has been shown to induce hyperglycemia when injected centrally. In recent years, two families of Bn-like peptides have been isolated from the mammalian brain: the gastrin releasing peptide (GRP) type and neuromedin B (NB) type. Distinct receptor subtypes with different mRNAs have also been identified for NB versus GRP/Bn using hybridization and receptor binding studies. It is thus possible that those two families of peptides may display distinct pharmacological profiles. The objective of the current experiment was to determine whether the NB-like peptides could also affect blood glucose levels. The peptide analogs utilized were Bn, NB-10, NAcNB-10 and NB-32 (0, 0.031, 0.062, 0.31, 0.62, 3.1 nmol/3 microliters; i.c.v.). Male rats, chronically implanted with 4th ventricular cannula, were injected with the various neuropeptide doses using the Latin square design. Blood samples were collected (120 microliters) from the tail immediately preceding and at 15, 30 and 60 min following peptide administration. Bn elevated glucose for over 60 min and this effect was maximal at 30 min. NB-10 and NAcNB-10 only slightly elevated plasma glucose. NB-32 elevated plasma glucose at all doses tested, the effect being evident up to 60 min at the highest dose. Our data indicate that at equimolar doses (0.31 nM) NB analogues elevate blood glucose with a lower efficacy than Bn (Bn > NB-32 > NB-10 > or = NAcNB-10). NB-32 appears more potent and efficacious than the other NB congeners used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234900 TI - Effects of the enkephalin analog (D-Met2,Pro5)-enkephalinamide on alpha melanocyte-stimulating hormone secretion. AB - We used the met-enkephalin analog (D-Met2,Pro5)-enkephalinamide (DMPEA) to investigate enkephalinergic control of alpha-melanocyte-stimulating hormone (alpha-MSH) secretion. Systemic (s.c.) administration of DMPEA elevated plasma titers of alpha-MSH in a dose- and time-related manner. Pretreatment with the opiate antagonist naltrexone had no effect on basal plasma levels of alpha-MSH but blocked DMPEA-induced alpha-MSH release. Treatment with a dose of naltrexone sufficient to block DMPEA-induced secretion of alpha-MSH had no effect on stress induced secretion of alpha-MSH. Although pretreatment with the dopamine receptor agonist apomorphine prevented DMPEA-induced alpha-MSH secretion, DMPEA had no effect on the synthetic activity of tuberohypophysial dopamine neurons as gauged by measuring the accumulation of 3,4-dihydroxyphenylalanine in the neurointermediate lobe (NIL) following administration of NSD-1015. In vitro treatment of isolated NILs with DMPEA resulted in a significant increase in alpha MSH release. Naltrexone completely blocked the stimulatory effects of DMPEA on alpha-MSH release in vitro. Our results indicate that DMPEA stimulates alpha-MSH secretion by acting directly through opiate receptors at the level of the NIL. PMID- 8234901 TI - MEN 10,573 and MEN 10,612, novel cyclic pseudopeptides which are potent tachykinin NK-2 receptor antagonists. AB - The activity and selectivity of MEN 10,573 and MEN 10,612, novel cyclic pseudopeptides which are selective tachykinin NK-2 receptor antagonists is described, as compared to that of previously characterized linear and cyclic compounds. For the NK-2 receptor, the activity of test compounds was investigated in the hamster isolated trachea (HT) and the endothelium-deprived rabbit isolated pulmonary artery (RPA), two preparations which are endowed with pharmacologically distinct forms of the NK-2 receptor. The novel cyclic pseudopeptides, MEN 10,573 and MEN 10,612 displayed very high affinity for the NK-2 receptor in the HT (pA2 8.66 and 9.06, respectively) which is higher than that observed in the RPA (pA2 7.31 and 7.41 for MEN 10,573 and MEN 10,612, respectively). The antagonism exerted by MEN 10,573 and MEN 10,612 was of competitive nature in both preparations. MEN 10,573 and MEN 10,612 also displayed competitive antagonism for NK-2 receptor-mediated responses in the rabbit bronchus (RB), rat vas deferens (RVD), circular muscle of the human colon (HUC) and ileum (HUI). In the RB, HUC and HUI, the potency of the novel cyclic pseudopeptides was comparable to that of MDL 29,913 and about 10-fold greater than that of L659,877. In the RVD however, the potency of MEN 10,573 MEN 10,612 or MDL 29,913 was similar to that of L659,877. In anaesthetized rats, i.v. injection of MEN 10,612 produced a selective and long-lasting blockade of the urinary bladder contraction produced by the i.v. injection of the NK-2 receptor selective agonist [beta Ala8]neurokinin A(4-10), without affecting the response to the NK-1 receptor selective agonist [Sar9]substance P sulfone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234902 TI - Comparison of the acute influence of neuropeptide Y and sympathetic stimulation on the composition of blood cells in the splenic vein in vivo. AB - The acute influence of exogenous transmitters and sympathetic nerve stimulation on the composition of blood cells in the splenic vein in relation to the splenic vascular effects was investigated in anaesthetized pigs. Intra arterial bolus injections of 720 pmol neuropeptide Y (NPY), 4.9 nmol noradrenaline (NA) and 20 nmol alpha,beta-methylene adenosine triphosphate (mATP) in the spleen were given and these doses caused arterial vasoconstriction in the same range, and increase in splenic venous haematocrit. NPY administration evoked a decrease in splenic venous blood flow and an unchanged leukocyte outflow from the spleen. mATP and NA, on the other hand, evoked increases in splenic venous blood flow and leukocyte outflow. Sympathetic nerve stimulation caused increases in haematocrit and leukocyte outflow in control pigs as well as in pigs with reserpine-induced depletion of tissue NA, although these effects, as well as the vascular effects, were significantly reduced after reserpine treatment. For comparison, the vasodilator calcitonin gene-related peptide increased leukocyte outflow without change in haematocrit. It is concluded that haematocrit and leukocyte concentration in the splenic venous blood are acutely modulated in different ways by vascular changes evoked by different sympathetic mediators. Furthermore, the capacitance function seems to be regulated by adrenergic and possibly purinergic transmission, whereas the non-adrenergic mediator NPY seems to be involved mainly in splenic arterial vasoconstriction. PMID- 8234903 TI - Noradrenergic and neuropeptide Y-immunoreactive nerves in the pancreatic islets of spontaneously hypertensive rats. AB - Innervation by noradrenergic (NA) and neuropeptide Y-immunoreactive (NPY-IR) nerves to the pancreatic islets and the arteries supplying the islets was studied by immunocytochemistry in 6- and 20-week-old spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). Since both the NA and NPY-IR nerves follow the intra- and interlobular small arteries and innervate the islets, we examined mainly the innervation of intra- and interlobular small arteries by these nerves. The NA nerve density of the intralobular small arteries supplying the islets was significantly more extensive in 6-week-old SHR than in age-matched WKY (7.11 +/- 1.78 vs. 4.04 +/- 0.71 x 10(-2) micron/micron2, P < 0.001), and the NPY-IR nerve density of the intralobular small arteries was significantly more extensive in SHR than in WKY at 6 weeks of age (7.68 +/- 2.28 vs. 5.69 +/- 0.91 x 10(-2) micron/micron2, P < 0.05). The interlobular small arteries branching off the intralobular small arteries in SHR were also more innervated by NA (P < 0.05) and NPY-IR nerves (P < 0.001) than by those in WKY at 6 weeks of age. At 20 weeks of age, the NA and NPY-IR nerve densities of the intra- and interlobular small arteries were higher in SHR than in WKY, and showed an increase with age in both strains. In addition, the islet innervation by these nerves in cross-section appeared denser in SHR than in WKY.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234904 TI - The primary structure of pancreatic polypeptide from a primitive insectivorous mammal, the European hedgehog (Erinaceous europaeus). AB - Pancreatic polypeptide (PP) has been isolated from extracts of the pancreas of the European hedgehog (Erinaceous europaeus) which is a representative of the order Insectivora, deemed to be the most primitive group of placental mammals. Pancreatic tissues were extracted in acidified ethanol and the peptide was purified chromatographically using a PP C-terminal hexapeptide amide specific radioimmunoassay to monitor purification. Two major PP-immunoreactive peptides were baseline-resolved following the final analytical reverse phase HPLC fractionation. Each was separately subjected to plasma desorption mass spectroscopy (PDMS) and gas-phase sequencing. The molecular masses of each peptide were similar: (I) 4237.6 +/- 4 Da and (II) 4238.2 +/- 4 Da. The full primary structures of each peptide were deduced and these were identical: VPLEPVYPGDNATPEQMAHYAAELRRYINMLTRPRY. The peptides were deemed to be amidated due to their full molar cross-reactivity with the amide-requiring PP antiserum employed in radioimmunoassay. The molecular mass (4233.8 Da) calculated from the sequence was in close agreement with PDMS estimates and the reason for the different retention times of each peptide is unknown at present. Hedgehog PP exhibits only 2 unique amino acid substitutions, at positions 1 (Val) and 19 (His), when compared with other mammalian analogues. PMID- 8234905 TI - Isolation and primary structure of a novel avian pancreatic polypeptide from five species of Eurasian crow. AB - Chicken pancreatic polypeptide is the prototype of the neuropeptide Y (NPY)/PP superfamily of regulatory peptides. This polypeptide was appended the descriptive term avian, despite the presence of some 8600 extant species of bird. Additional primary structures from other avian species, including turkey, goose and ostrich, would suggest that the primary structure of this polypeptide has been highly conserved during avian evolution. Avian pancreatic polypeptides structurally characterised to date have distinctive primary structural features unique to this vertebrate group including an N-terminal glycyl residue and a histidyl residue at position 34. The crow family, Corvidae, is representative of the order Passeriformes, generally regarded as the most evolutionarily recent and diverse avian taxon. Pancreatic polypeptide has been isolated from pancreatic tissues from five representative Eurasian species (the magpie, Pica pica; the jay, Garrulus glandarius; the hooded crow, Corvus corone; the rook, Corvus frugilegus; the jackdaw, Corvus monedula) and subjected to structural analyses. Mass spectroscopy estimated the molecular mass of each peptide as 4166 +/- 2 Da. The entire primary structures of 36 amino acid residue peptides were established in single gas-phase sequencing runs. The primary structures of pancreatic polypeptides from all species investigated were identical: APAQPAYPGDDAPVEDLLRFYNDLQQYLNVVTRPRY. The peptides were deemed to be amidated due to their full molar cross-reactivity with the amide-requiring PP antiserum employed. The molecular mass (4165.6 Da), calculated from the sequences, was in close agreement with mass spectroscopy estimates. The presence of an N-terminal alanyl residue and a prolyl residue at position 34 differentiates crow PP from counterparts in other avian species.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234906 TI - Expression of functional receptors for vasoactive intestinal peptide in freshly isolated and cultured gastric muscle cells. AB - Vasoactive intestinal peptide (VIP) receptors were characterized in freshly isolated and cultured smooth muscle cells from guinea pig stomach by radioligand binding and by measurement of relaxation in single isolated and cultured cells. 125I-VIP bound to both freshly isolated and cultured muscle cells: binding was rapid, specific, saturable and temperature-dependent, and was inhibited in a concentration-dependent fashion by VIP, VIP10-28, PHI and secretin, in this order. Competition curves for VIP could be resolved into high- and low-affinity components, yielding similar binding constants in freshly isolated and cultured cells (high-affinity Kd 0.11 and 0.22 nM; low-affinity Kd 59 and 37 nM; high affinity binding sites: 1183 and 1021 per cell, representing about 1% of total binding sites). VIP10-28 inhibited 125I-VIP binding completely and acted as potent competitive antagonist of VIP-induced relaxation (Ki 0.5 nM). PHI and secretin, however, inhibited partly 125I-VIP binding: the pattern of inhibition implied that VIP interacts with VIP-preferring receptors that are recognized by PHI and secretin as well as with VIP-specific receptors. The pattern of binding is consistent with recent evidence indicating that VIP activates two signalling pathways, a VIP-specific, nitric oxide/cGMP-dependent pathway and a common cAMP dependent pathway shared by all three peptides. PHI and secretin were relatively more potent as relaxant agents than as inhibitors of 125I-VIP binding raising the possibility that PHI and secretin could interact additionally with PHI- and secretin-preferring receptors in mediating relaxation. PMID- 8234907 TI - Cyclooxygenase products mediate the cutaneous vasodilation induced by endothelin 1 in humans. AB - Intradermal injection of endothelin-1 (ET-1) causes vasoconstriction (pallor) at the injection site, surrounded by a larger area of vasodilation (flare) in humans. Some of the vasomotor responses to ET-1 are thought to be mediated by prostaglandins. In the present study, we investigated the involvement of cyclooxygenase-derived products of arachidonic acid metabolism on the cutaneous vasomotor responses to ET-1. Ten normal subjects (25-44 years) were studied after treatment with either indomethacin (50 mg t.i.d.) or placebo according to a double blind cross-over design. Five doses of ET-1 (5 x 10(-5) to 5 x 10(-1) pmol) were injected intradermally 2 h after the last dose of indomethacin or placebo. Pallor and flare areas measured by planimetry 15 min after the injection were analyzed to evaluate cutaneous vasomotor responses to ET-1. ET-1 induced dose-dependent pallor and flare responses that were significant at the dose of 5 x 10(-3) pmol or greater. Indomethacin did not affect the ET-1-induced pallor but significantly shifted to the right the flare dose-response curve to ET-1. The inhibition of the flare response to 5 x 10(-1) pmol ET-1 was 58.9 +/- 8.5%. These results indicate that the cutaneous vasodilation induced by intradermal injection of ET-1 is mediated by the release of vasodilating cyclooxygenase products. PMID- 8234908 TI - Immunoreactive neuropeptide Y (NPY) in plasma and platelets of rat and mouse strains and human volunteers. AB - Immunoreactive-neuropeptide Y (i-NPY) is present in platelets of rats, and has recently been demonstrated to be authentic rat NPY based on its amino acid sequence. This potent vasoconstrictor and putative smooth muscle mitogen is released during platelet activation, suggesting a role in platelet-vascular interactions. We have now extended this work to several strains of rats and mice, and humans of both sexes. Among mice, strains in which NPY mRNA has been demonstrated in megakaryocytes have markedly higher levels of i-NPY (0.63-1.11 pmol/ml in NZB/B1NJ, NZBWF1/J, BXSB/MpJYaa, BALB/cJ) in platelet rich plasma (PRP) than other strains (DBA/2J, CBA/J, C3H/HeJ, MRL/MpJ-lpr, C57BL/6J; each < 0.02 pmol/ml). In rats, high content of i-NPY was observed in PRP and platelets of all strains examined (Sprague-Dawley, Wistar, Wistar Kyoto). i-NPY level was 30.6, 3.7 and 10.1 pmol/ml in PRP of the three strains, respectively. In humans, low levels of i-NPY occur in plasma and platelet fractions compared to rodents (0.069 and 0.048 pmol/ml in male and female PRP, respectively), but they, too, have greater i-NPY in platelet rich plasma and platelets than in platelet poor plasma. Assuming this is authentic NPY, platelet-derived NPY might have a role in pathophysiological states involving activation of platelets in humans. PMID- 8234909 TI - A proposed bovine neuropeptide Y (NPY) receptor cDNA clone, or its human homologue, confers neither NPY binding sites nor NPY responsiveness on transfected cells. AB - Receptors with seven transmembrane domains (7TM) constitute a large family of structurally and functionally related proteins which respond to various types of ligands. We describe here the cloning and expression of a human 7TM receptor, denoted hFB22 (human Fetal Brain 22), which is the homologue (92% amino acid identity) of a bovine receptor (LCR1) reported by others to bind neuropeptide Y (NPY) with a pharmacological profile of the Y3 receptor subtype. However, upon expression in COS1 (confirmed by Northern analysis), COS7 or CHO-K1 cells, the hFB22 receptor did not confer specific 125I-Bolton-Hunter-NPY, 3H-propionyl-NPY or 125I-peptide YY (PYY) binding sites, in either intact cells or in membrane preparations. Similarly, cells transfected with the corresponding bovine clone (LCR1) did not show specific NPY/PYY binding exceeding that resulting from endogenous binding sites; mock-transfected COS7 cells, used frequently for heterologous expression of receptors, were found to have endogenous specific 125I NPY binding sites (Bmax = 112 fmol/mg protein; Kd = 0.25 nM). Moreover, the hFB22 transfected cells, when compared to control transfected cells, did not display de novo NPY- or PYY-induced second messenger responses, i.e., (1) inhibition of forskolin-stimulated cAMP accumulation or (2) 45Ca2+ influx. The presence of hFB22 mRNA was detected in several human neuroblastoma cell lines, none of which was found to express Y3-like NPY binding sites. hFB22 displays 39% amino acid sequence identity (in the transmembrane regions) to the human interleukin-8 receptor, and 32-36% amino acid identity to the human receptors of angiotensin II, bradykinin, and n-formylpeptide, but only 23% amino acid identity to the previously described human NPY/PYY receptor of the Y1 receptor subtype. Our results show that hFB22 and LCR1 do not encode NPY receptors, and their true ligand(s) remains to be identified. PMID- 8234910 TI - Changes in enkephalin immunoreactivity of sympathetic ganglia and digestive tract of the cat after splanchnic nerve ligation. AB - The aim of the present study was to analyze changes in the enkephalin immunoreactivity of sympathetic prevertebral ganglia coeliac plexus and inferior mesenteric ganglion) and intestinal tract (myenteric plexus and external muscle layers) in cats 2 days after left thoracic splanchnic nerve ligation, using radioimmunoassay and immunohistochemical techniques. Specific polyclonal antibodies directed against methionine- and leucine-enkephalin were used. The nerve ligation led to a considerable increase in the enkephalin immunoreactivity in the cranial part of the ligated nerves. This finding confirms the presence, in the cat, of an enkephalin output originating from thoracic spinal structures which are probably enkephalin-containing preganglionic neurons. In prevertebral ganglia the nerve ligation induced a marked decrease in the enkephalin immunoreactivity, which was probably due to the interruption of thoracic enkephalin efferents projecting towards both the coeliac plexus and the inferior mesenteric ganglion. In the digestive tract, the nerve ligation depressed the methionine-enkephalin immunoreactivity only in the gastro-duodenal region, and had no effect on the ileo-colonic region. The results of the present study add to the growing evidence that the sympathetic nervous system is involved in regulating the enteric enkephalinergic innervation, which is probably involved in controlling the intestinal motility. PMID- 8234911 TI - A high-molecular IGF-2 immunoreactive peptide (pro-IGF-2?) in the insulin cells of the islets of Langerhans in pancreas of man and rat. AB - Histopathologically normal pancreatic parenchyma from 12 adult men and women, as well as that from 14 adult rats (Sprague-Dawley and Wistar strains), were investigated immunohistochemically with a mouse monoclonal antibody, raised against recombinant human pro-IGF-2. The antiserum showed no crossreactivity with insulin; IGF-1 had 0.1% of the reactivity of IGF-2. The immunohistochemical observations were checked by means of a radioimmunoassay (RIA), based on the same antibody, of an extract of a sample of one of the human pancreatic glands. Analogous investigations for insulin were made in parallel, using polyclonal insulin antisera. A high-molecular (12 kDa) IGF-2-like peptide was found in the islets of Langerhans, being localized to the insulin cells. These cells were identified as beta-cells by immunohistochemistry with insulin antisera on adjacent paraffin sections. From observations made by means of acid-gel chromatography, the peptide was tentatively supposed to represent either pro-IGF 2, or a partially processed form of it. PMID- 8234912 TI - Morphometry of gastric endocrine cells in hypergastrinemic patients treated with the somatostatin analogue octreotide. AB - The aim of the study was to evaluate whether treatment with 200 micrograms/d of the somatostatin analogue octreotide (SMS 201-995) for three months can influence the trophic action exerted by hypergastrinemia on endocrine cells of the oxyntic mucosa, a condition potentially leading to hyperplasia and carcinoid tumors. Endocrine cells were morphometrically investigated in Grimelius silver stained sections of endoscopic biopsies of oxyntic mucosa collected from 13 hypergastrinemic patients with Zollinger-Ellison syndrome (ZES) (n = 5), antral G cell hyperfunction (AGCH) (n = 4) and atrophic gastritis type A (AG-A) (n = 4) before and after 3 months treatment and 3 months after drug discontinuance. The treatment induced a reduction of the volume density (P < 0.015), profile cross sectional area (P < 0.05) and number of cell profiles per unit area (P < 0.015) of argyrophil cells. A rebound of all these parameters was observed 3 months after drug withdrawal with values usually exceeding those at the entry, except in cases of AG-A. The patients' plasma gastrin concentrations presented similar variations showing a significant relation with all morphometric parameters of argyrophil cells. Also, the cell content in alpha subunit of human chorionic gonadotropin was related to the plasma gastrin levels, a finding confirming the close gastrin dependence of the expression of this protein by oxyntic endocrine cells. No significant changes were observed in mucosal somatostatin D cells. These results indicate that variations in circulating gastrin levels are the most likely factor responsible for the hypotrophic effect of octreotide on oxyntic argyrophil cells (mostly corresponding to the ECL cells) of hypergastrinemic patients. PMID- 8234913 TI - The paradoxical lack of interspecies correlation between plasma concentrations and chemical carcinogenicity. AB - Several chemicals have been shown, within the limits of epidemiology, to be thus far not carcinogenic in humans at systemic exposure levels similar to those associated with carcinogenicity in rodents. The data are discussed in terms of mechanisms which appear to operate in rodents but not in humans. Pharmacokinetic methods for interspecies comparisons must always be validated by a complementary scrutiny of the pharmacodynamic processes involved, i.e., by considering the responsiveness of the species in question to a given systemic concentration of xenobiotic. For carcinogenesis this validation step is usually beyond reach, due to the time frame for the onset of cancer in humans. For these reasons it is argued that, in the absence of knowledge of mechanisms, there is usually no scientific basis for using the concentrations of a xenobiotic that occur in body fluids or tissues during a rodent carcinogenicity test to make a quantitative carcinogenic risk assessment in humans. PMID- 8234914 TI - The science of toxicology and its relevance to MCS. AB - The definitions and current practice of multiple chemical sensitivity (MCS) are considered in relation to the fundamental principles of toxicology. The salient problem with MCS is that there is no consistent and specific effect from exposure to any specific chemical. This does not allow for any objective test for any disease entity which might be caused by the chemicals as indicated by the theory of MCS. The effects of exposure to chemicals as defined today as MCS are subjective and no report is available to convincingly demonstrate that these effects would not have occurred merely by chance. It is concluded that (1) MCS, as defined, does not allow specificity or consistency for biological reactions to the effects of chemicals; (2) MCS does not provide a testable hypothesis; (3) MCS is contradictory to the fundamental principles of toxicology; (4) current testing procedures for MCS are so subjective that they are useless; (5) there is no evidence that the responses attributed to MCS are any other than those that would occur merely by chance; and (6) the MCS literature attaches an emotional bias to chemicals. PMID- 8234915 TI - Medical hypothesis and medical practice: autointoxication and multiple chemical sensitivities. AB - Searching for simple explanations for ill health is as normal as human nature itself. In every era people have suffered varieties of symptoms not readily explained by "known" diseases of the day. To provide explanations and treatment approaches, individuals with symptom complexes were frequently provided a unifying disease label. At the turn of the 20th century, that label was autointoxication. Today, it is multiple chemical sensitivities (MCS). Autointoxication, a popular turn-of-the-century phenomenon illustrates that absent common, reproducible, definable, and measurable findings and consistent, objective endpoints, newly named fad disorders have not survived long term. Autointoxication was a disorder very similar in manifestations to MCS. It was, we now know, a pseudodisease, the treatment of which was based upon an unproven hypothesis. If MCS cannot withstand the probing of scientific investigation, it too will disappear as a diagnosis. PMID- 8234916 TI - Environmental illness and misdiagnosis--a growing problem. AB - A fringe group of medical practitioners called clinical ecologists believes that hypersensitivity to common foods, chemicals, and organisms can disrupt the immune system and lead to diverse medical or psychiatric problems. They believe this condition, frequently referred to as environmental illness (EI), can be diagnosed on the basis of a patient's history of exposure, and the results of provocation testing and elimination diets. They advise treating the condition with avoidance of the offending agent(s), special diets, and symptom neutralization. Other treatments are often recommended. Clinical ecology beliefs and practices have been criticized by mainstream medical practitioners who have urged that EI not be recognized as a clinical syndrome. Research has shown that individuals receiving a diagnosis of EI frequently have common psychiatric or medical disorders, which are usually unrecognized and untreated. Thus, the clinical ecologists are misinterpreting common signs and symptoms of illness and failing to prescribe appropriate and proven therapies. The advice and recommendations of a clinical ecologist can lead to iatrogenic social and occupational disability. PMID- 8234917 TI - Multiple chemical sensitivity: controlled scientific studies as proof of causation. AB - Multiple chemical sensitivity is an environmental illness that demands exacting methods of diagnosis. Proposed associations between symptoms and specific substances, whether to one substance or to multiple chemicals, need to be critically examined through adequately designed scientific studies. Appropriate methods for controlled scientific study of adverse reactions to chemicals are discussed as well as the Food and Drug Administration's (FDA's) experience with aspartame as an example of the need for controlled scientific studies to refute or confirm anecdotal evidence. Since 1986 the FDA has received reports of 265 cases of epileptic seizures temporally associated with the ingestion of aspartame. Information obtained from the complainants' medical records as well as data on consumption patterns, temporal relationships, and challenge tests do not support the claim that the occurrences of the seizures are linked to consumption of aspartame. In addition, two double-blind, placebo-controlled crossover studies failed to demonstrate an association between epileptic seizures in children and adults and the ingestion of aspartame. PMID- 8234918 TI - Double-blind provocation chamber challenges in 20 patients presenting with "multiple chemical sensitivity". AB - A clinical algorithm was used to discriminate verifiable chemical sensitivity from psychological disorders in patients referred for evaluation of polysomatic symptoms attributed to hypersensitivity to workplace and domestic chemicals. These patients believed that they were reactive or hypersensitive to low-level exposure to multiple chemicals. Some had previously been evaluated and managed by the tenets of "clinical ecology" and diagnosed as having "multiple chemical sensitivity." Double-blind provocation challenges with an olfactory masker were performed in an environmental chamber on each of 20 patients. A variety of chemicals was employed, one or more per subject, dependent on individual clinical history. Clean air challenges with the olfactory masker were used as placebo or sham controls. As a group, probability analyses of patient symptom reports from 145 chemical and clean air challenges failed to show sensitivity (33.3%), specificity (64.7%), or efficiency (52.4%). Individually, none of these patients demonstrated a reliable response pattern across a series of challenges. Implications for future research in assessment methodology incorporating neurophysiologic and neurobehavioral measures are discussed. PMID- 8234919 TI - Immunological issues in "multiple chemical sensitivities". PMID- 8234920 TI - Safety assessment of hydrogenated starch hydrolysates. AB - Hydrogenated starch hydrolysates (HSH) are mixtures of polyhydric alcohols such as sorbitol, maltitol, and higher-order sugar alcohols. They are important food ingredients because of their sweetness, low cariogenic potential, and useful functional properties. These traits permit HSH products to be used as viscosity or bodying agents, humectants, crystallization modifiers, and rehydration aids. A substantial body of safety information is available for HSH products and their individual chemical components. Based on this information, the substances have received favorable evaluations from international expert safety organizations such as the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and European Community's Scientific Committee for Food. This same information has been submitted to the United States Food and Drug Administration (FDA) as part of the petitioning process to affirm the generally recognized as safe (GRAS) status of these substances. Some of the animal feeding studies important to a full safety assessment for HSH substances, while long available to international safety expert organizations and governmental organizations, have never been published in the literature. Three of these studies, i.e., a chronic (24-month) feeding study, a multigeneration reproduction study, and a teratology study, are reported on this article, together with metabolic information. The results of this evaluation establish HSH substances as safe food ingredients. PMID- 8234921 TI - Recent developments in alcoholism:ethnicity. AB - Advances in alcohol and ethnicity epidemiology during the past decade include greater emphasis on intraethnic variation, factors influencing drinking behavior, and building conceptual models. Despite progress, most research continues to focus on ethnicity/race as a demographic variable and few studies investigate ethnic hypotheses. Although prevention and treatment of alcoholic disorders in ethnic groups have been dealt with extensively in the clinical and ethnographic literature, little rigorous research has been conducted. The key issue in this area involves the extent to which prevention and treatment programming needs to be ethnically responsive in order to be effective. The very little empirical literature that exists on this topic suggests that answers will not be simple and will vary according to aspects of acculturation status and ethnohistorical factors. The most important development during the past 10 years has undoubtedly been the federal regulatory encouragement of research on the major ethnic groups of the United States. PMID- 8234922 TI - Recent developments in alcoholism:the workplace. AB - We examine the results of recent research that considers the relationship of the work environment to drinking behavior and problems. We bring the concept of culture to the foreground in this discussion, and show that much of what alcohol researchers term as work-related risk factors are, in effect, best understood as aspects or dimensions of cultural processes which operate in complex organizations. We identify and review four perspectives on alcohol use in occupational contexts that highlight these cultural processes at work: (1) the development of organizational norms with respect to drinking, definitions of problem drinking, and mechanisms of social control; (2) working conditions that produce in some workers experiences of powerlessness, alienation, or stress which are in turn alleviated by drink; (3) the interaction of cultural factors external to the workplace--regional/national, community, ethnic, and religious--with organizational cultural norms which affect alcohol values, beliefs, and behaviors; and (4) processes that underlie the development of occupationally based drinking subcultures. PMID- 8234923 TI - Recent developments in alcoholism:public drinking. AB - Drinking in bars and other public establishments has been a relatively neglected subject of research, despite its epidemiological significance. This chapter briefly summarizes the findings of observational studies in bars and taverns, as well as general population surveys on the correlates of tavern patronage. Recent developments in the control of public drinking are described. It is argued that greater attention should be placed on prevention programs that focus on drinking environments. In particular, the emergence of server intervention programs represents a promising new mechanism for the prevention of impaired driving and other alcohol-related problems. PMID- 8234924 TI - Recent developments in alcoholism:the liver. AB - The past decade has witnessed major gains in our understanding of the pathogenesis and therapy of alcoholic liver disease. The molecular biology of alcohol-metabolizing enzymes is well understood. Older concepts of liver injury, e.g., hypermetabolism, generation of free radicals, mitochondrial and microtubular dysfunction, and impairment of liver regeneration by ethanol, have been studied in greater detail. The fibrotic response to alcoholic liver injury has been explored, revealing complex interrelationships between the nonparenchymal cells of the liver and showing the importance of cytokines in regulating these cells. New mechanisms of injury have been appreciated, most prominently the association between hepatitis C infection and alcoholic liver disease, and the formation of protein-acetaldehyde adducts in the liver of alcohol-fed subjects. A new animal model of alcoholic liver injury, the alcohol infusion rat model developed by French and Tsukomoto, promises to provide a relatively simple model for researchers. The clinical management of alcoholic liver disease continues to evolve. Focal fatty change is recognized as a variant of alcoholic fatty liver. Nonalcoholic steatohepatitis has been described as a mimic of alcoholic liver disease, and may provide insight into the mechanisms of perivenular liver injury. The presence of perivenular fibrosis may predict at an early stage which patients are at risk for serious liver injury. Nutritional and corticosteroid therapy of alcoholic hepatitis are now established. Other therapies such as propylthiouracil, glucagon plus insulin infusion, and colchicine have been studied in large trials. Alcoholic liver disease can now be treated in selected cases by liver transplantation. PMID- 8234925 TI - Recent developments in alcoholism:genetic transmission. AB - This chapter is an overview of developments in the study of genetic factors in vulnerability in alcoholism. The focus is on recent developments, including heritability studies in twins and transmission studies in families, both of which have continued to reveal evidence for a substantial role for genetic factors but also for etiologic complexity and variation in vulnerability across generations and across cultures. Studies are discussed which utilized psychophysiological and neurochemical markers for alcoholism for analysis of genetic association, transmission, and linkage. These markers include the low P300 event-related potential, sensitivity to ethanol's intoxicating and euphoric effects, platelet adenylate cyclase, and neurotransmitter metabolite concentrations. Although it is highly likely that many alcoholism-associated physiologic phenotypes are secondary traits, these approaches have increased the specificity of genetic analyses and genetic analyses are clarifying their relationship to alcoholism. For example, early efforts to study, in families, the co-occurrence of the P300 marker and alcoholism have yielded results indicating that the P300 abnormality precedes significant exposure to alcohol and that relatives of alcoholics are more likely to have this trait. In the area of animal models, two nonhuman primate species, the vervet monkey and the rhesus macaque, were shown to willingly consume alcohol to intoxicating blood levels. Also, linkage studies using the quantitative trait locus (QTL) mapping strategy were attempted for phenotypes relevant for alcoholism. The QTL strategy is theoretically capable of identifying determinant genes which contribute only a small portion of the variance. In human linkage studies, a genetic association was found to the DRD2 dopamine receptor. The DRD2 finding generated controversy, as a number of other groups failed to replicate it, and also focused attention on the advantages and pitfalls of the population association approach for detecting genes influencing behavior. The relationship of the alcohol metabolic gene variants to alcoholism was clarified by the finding that functional variants of alcohol and aldehyde dehydrogenases can act additively to determine vulnerability to alcoholism. PMID- 8234926 TI - Recent developments in alcoholism:immunological aspects. AB - The association between chronic ethanol use and a predisposition to infection and increased severity of infection has been recognized by clinicians for many years. Clinical studies over the last century have substantiated individual clinical observations. Numerous studies have indicated that alcoholics are more susceptible to pulmonary infections and do not respond to treatment as well as nonalcoholic patients. A diminished ability to clear bacteria after chronic or acute ethanol treatment has also been demonstrated in a variety of experimental animals. Within the last few years a number of investigators have attempted to elucidate the mechanisms responsible for the apparent impairment of the immune system by either chronic or acute ethanol treatment. Alcohol has been reported to have adverse effects on all major components of the immune system. Ethanol affects the number of immunocompetent cells as well as the function of the remaining cells. In this review the effect of acute ethanol intoxication or chronic ethanol use on leukocyte and lymphocyte numbers, alteration of cellular function, and ability of the cells to arrive at the site of infection are addressed. PMID- 8234927 TI - Recent developments in alcoholism:biobehavioral correlates. AB - A literature review was conducted from 1940 through 1992 on the general topic of alcohol-induced hangover. We focused on the physiological, behavioral, and performance decrements demonstrated hours after a physiologically relevant dose of alcohol was administered in both humans and animals. The purpose of this report was threefold: (1) to review and report both previously published and new data from our laboratory concerning the "after effects" of acute high-dose alcohol administration; (2) to highlight the similarities between and usefulness of data generated from both human and animal studies; and (3) to propose that the hangover phenomenon may be the result of one of the three following hypotheses: (a) an opponent-process physiological rebound from alcohol's immediate, intoxicating effects, (b) some toxic reaction to alcohol or its metabolic by products, or (c) to the circadian dysrhythmia induced by the immediate intoxicating effects of alcohol. PMID- 8234928 TI - Recent developments in alcoholism:anthropology. AB - During the past few decades, the volume of literature on anthropological studies of alcohol has doubled every five years. New topics are being investigated, and a variety of new methods, including quantitative analysis, have been integrated with the proven qualitative approaches. This chapter reviews work since 1980, illustrating both continuity and change, as well as describing relevant developments in terms of practical applications of the findings. Special emphasis is given to recognition and interpretation of cross-cultural variation, qualitative data and emphasis on context, and critical discussion of concepts, models, and theories, as many more, better trained, investigators address new as well as old topics and relate their studies to issues of policy and action. PMID- 8234929 TI - Recent developments in alcoholism:typologies in women. AB - Historically, there has been considerable interest in developing alcoholic typologies with the belief that typologies would facilitate treatment development and stimulate meaningful research. Most of the existing typologies can be classified into one of two broad camps: the behavioral typologies and the family history/genetic typologies. Recent work has focused on the integration of these two strategies into comprehensive typological systems. This chapter provides an overview of the major typologies, noting their various strengths and weaknesses. Common weaknesses among existing typologies are that female alcoholics have not been adequately studied and/or are stereotyped. Behavioral heterogeneity within the female alcoholic population is discussed and suggestions for typological development are noted. PMID- 8234930 TI - Reducing the desire to drink. Pharmacology and neurobiology. AB - The past decade has witnessed major advances in understanding of neural functioning and neurobiological bases of alcohol consumption. Concurrent with this, a range of exciting investigations have been conducted on pharmacologic agents that may curb drinking behavior. Research is reviewed on several promising medications influencing neurotransmitter and endocrine systems with particular attention to the serotonergic and opioid systems. Following this overview, recommendations are offered regarding research methodology to support future pharmacotherapy trials. PMID- 8234931 TI - Developments in alcoholism treatment. AB - Alcohol treatment systems expanded and diversified considerably over the past decade. This reflects adaptation to a variety of forces, including developments in national health care financing and policy, changes in other health care systems with which alcohol treatment had strong ties, the more diffuse effects of social movements and a "drying trend" in American public opinion, as well as agitation by advocacy and provider groups within the alcohol field. Drawing on national monitoring data, this chapter reviews developments at the levels of financing policy, organizations, client populations, and treatment modalities, documenting expansion in private sector alcohol treatment units, a growing emphasis on providing outpatient treatment, a merger between services for alcohol and drugs at the organizational and conceptual levels, increases in service delivery to coerced populations, as well as demographic change in alcohol treatment caseloads during the 1980s. PMID- 8234932 TI - Recent developments in alcoholism:behavioral treatment. AB - Behavioral approaches have made significant contributions to understanding the etiology and treatment of alcohol dependence. The present chapter provides a brief overview of changes that have occurred within this perspective over the past decade. Significant changes at a theoretical level have included a move toward mediational cognitive-behavioral models of etiology and maintenance, integrative multivariate models, and an empirical, developmental model based on the natural history of the drinking and alcohol problems. Significant changes in clinical practice have included an expanded role of assessment within the context of a biopsychosocial model of alcoholism, an increased focus on secondary prevention and brief intervention with less-alcohol-impaired individuals, a focus on "harm reduction" as a goal of such interventions, the development of a stepped care approach to treatment with a reliance on patient-treatment matching, an increased emphasis on the maintenance of therapeutic gain and relapse prevention, and the incorporation of behavioral and cognitive principles into self-help programs. PMID- 8234933 TI - Recent developments in alcoholism:pharmacological treatment. AB - Neurobiological research over the past decade has greatly expanded our knowledge of alcohol's interaction with the nervous system and promises significant improvement in pharmacological treatment for all phases of alcoholism. Although no new treatment has yet become standard clinical practice, over the next decade benzodiazepine receptor antagonists and inverse agonists should improve treatment of acute alcohol intoxication, adrenergic agents and anticonvulsants should improve treatment of acute alcohol withdrawal, and serotonin uptake blockers and mu opiate receptor antagonists should improve long-term treatment of alcohol dependence. Future clinical research should reduce the current lack of knowledge on pharmacological treatment of alcoholism in special populations such as women, the elderly, and those with concurrent psychiatric diagnoses. PMID- 8234934 TI - Recent developments in alcoholism: inpatient treatment. AB - The historical role of inpatient treatment for alcoholism is reviewed in terms of its advantages and disadvantages. The factors that have forced a change in the utilization of inpatient treatment include increasing recognition of the heterogeneity of alcoholic patients, negative outcome studies, and cost containment efforts. The clinical domains that warrant inpatient treatment are outlined, and decisions of treatment placement are necessarily guided by the factors of acuteness, ability, safety, and stabilization. PMID- 8234935 TI - Recent developments in alcoholism:psychology. AB - During the past decade, the concerns of psychological research within alcohol studies have been far ranging and even within the areas of personality and neurocognition, the literature is voluminous and many faceted. This chapter focuses on aspects of the personality system that appear to increase vulnerability to excessive alcohol or other drug use or the experience of use related problems as well as issues related to temporal stability and change in intrapersonal development and drinking behaviors. The bulk of the neurocognitive literature reviewed deals with the impairments that accrue as a direct or indirect consequence of excessive alcohol consumption. While the individual empirical studies are often, by necessity, narrow in focus, the import of intrapersonal sources of vulnerability and the processes and mechanisms through which these may drive and moderate alcohol and other drug use behaviors and outcomes will be more fully understood within the context of conceptual models and research approaches that are complex, multilevel, and integrative. PMID- 8234936 TI - Recent developments in alcoholism:psychodynamic approaches. AB - A review of the substance abuse literature in the 1980s focuses on deficits in self-regulating capacities, in self-care functions, and in other vulnerabilities of the self, such as an inappropriate use of defenses and problems in intimacy. Various models which attempt to explain these behaviors are described, as well as techniques of dynamic psychotherapy for substance abuse, which emphasize the building of ego functions and the internalization of self-preservation functions. Commonalities and differences of substance abuse with other intensely driven compulsive behaviors (e.g., pathological gambling or certain sexual behaviors) are also considered. PMID- 8234937 TI - Dealing with alcohol problems in the workplace. AB - The employee assistance program (EAP) is the principal vehicle for intervention in the workplace directed at employees with alcohol problems. As broad-gauged programs, EAPs represent "mainstreaming" of alcohol-problem concerns into health care management. However, alcohol problems are defined broadly within EAPs, going beyond other definitions of alcohol dependence. The historical context is important for understanding EAPs and their dynamics. External environmental changes have impacted EAPs considerably over the past decade. Particularly important have been the increased prominence of employee drug abuse, managed care, and increasing prevalence of dual diagnoses. Results of research about EAPs conducted over the past decade are reviewed. The persistent issue of EAP efficacy is considered, highlighting the variable structure of EAPs and the multiplicity of motives for their voluntary adoption. Prospects for continuing research are considered in light of problems in research funding, the ambiguous identity of EAPs, and the practical orientations of those who might facilitate such research. PMID- 8234938 TI - Recent development in alcoholism:sociology. AB - This chapter reviews several domains of sociological research conducted in the field of alcohol studies during the last decade. The domains are organized within the sociostructural, social constructionist, and distribution of consumption theoretical perspectives. Discussions of the current state of the field and definitional and measurement issues are also included. Within the sociostructural perspective, sociocultural theories, social deviance theories, age, gender, ethnic, and occupational group differences, and other related deviant behaviors are examined. Several examples of applications of social constructionist theory are included covering the new temperance movement, the Mothers Against Drunk Driving movement, Alcoholics Anonymous, and the Children of Alcoholics movement. In the section on distribution of consumption models, alcohol availability and drunk driving control policies are discussed. There are several sociopolitical issues that need to be addressed in the next decade and further collaboration across disciplines is warranted. PMID- 8234939 TI - Children and alcohol. AB - The literature on what, when, and how young children learn about alcohol and drinking is critically reviewed and key issues are highlighted. Available evidence points to recognition of alcohol, identification of drinking norms and rules, formation of attitudes toward drinking and drinkers, and appreciation of some of the consequences of alcohol consumption as early as the preschool years. Understanding of drinking motives appears to require a level of cognitive development that is not present until about 10 to 12 years of age, although there is a dearth of research on this probably crucial determinant of later decisions to drink. Among the factors involved in the acquisition of alcohol cognitions and dispositions are observational learning via families, media, and peers as well as personal drinking experience. Salience of alcohol and its management in the child's immediate environment appears to be an important moderator/mediator of learning. Application of relevant theory to understanding the development of the meaning of alcohol and drinking in preadolescent children is seen as underdeveloped at this time. However, there are a number of potentially fruitful avenues to pursue which could have important implications for future education and prevention efforts. PMID- 8234940 TI - Recent developments in alcoholism:family systems. AB - Over the past decade, research on alcohol and family systems has made notable advances, as reflected by major publications. Concepts elucidated in these writings encompass issues of (1) risk and vulnerability (resiliency, vulnerability, at-risk groups, mediating moderating risk factors, multifactorial determinants, and predisposing and precipitating factors); (2) family process (regulatory mechanisms, homeostasis, morphogenesis, systemic maturation, developmental distortion, family rituals, family identity, and deliberateness); (3) drinking context (sobriety-intoxication cycle, steady-episodic drinking, and alcohol expectancies); (4) alcohol controls (coercive controls, informal drinking controls, and formal controls); and (5) family response to alcoholism (codependence, denial, and definitional careers). In the following review of some of the major contributions to this research tradition, we pay particular attention to how alcohol researchers have refined many of these concepts in light of new empirical data. PMID- 8234941 TI - Recent developments in alcoholism:gender issues. AB - Study of women and alcohol may involve a female/male comparison or comparison among subgroups of women. Epidemiological study over the last decades has not supported the hypothesis of convergence of male and female drinking rates, but changes in women's drinking patterns have occurred. Problem drinking and alcohol abuse in men and women are compared: genetics, vulnerability to liver damage, age at onset, spouse's drinking, use of other drugs. A critical issue is the public/private nature of drinking and the consequences which follow from that. Major antecedents to female problem drinking appear to be difficulties in impulse control, depression, and the earlier appearance of other diagnostic syndromes such as eating disorders or phobia. The consequences of problem drinking and alcoholism--interpersonal, legal, occupational, and medical--are discussed. In spite of the increase in gender-related research reports, women remain "the second sex" in theory development and diagnostic definitions. PMID- 8234942 TI - Environmental fate and effects of dicamba: a Canadian perspective. AB - Literature on the environmental fate and effects of the benzoic acid herbicide dicamba was reviewed to provide a scientific basis to derive Canadian Water Quality Guidelines. Included in the review was information on the uses and production of dicamba, its physical and chemical properties, environmental monitoring data in Canadian surface water and groundwater, soils, sediments, and biota, and its environmental degradation, persistence, and fate. Through monitoring, dicamba has been detected in less than 8% of surface-water samples to a maximum concentration of 13 micrograms.L-1, while 2% of groundwater samples were positive up to 517 micrograms.L-1. Only one study that analyzed sediments (with no detections) and no field studies that investigated residues in biota were found. Microbial degradation is the most important process governing the dissipation of dicamba in aquatic and soil environments. Photolysis, hydrolysis, volatilization, adsorption to sediment, and bioconcentration are not expected to be significant removal processes, based on limited environmental fate data. The half-life of dicamba in water is < 7 d, although residues have been detected in surface-water supplies in Alberta more than 6 mon after application. The literature reports the half-life in soils ranges from 4 to 555 d; however, < 12 wk would be typical under Canadian conditions. High moisture and temperature, and other conditions that favor microbial degradation, would likely reduce the half life to < 4 wk. The principal soil and plant metabolite is 3,6-dichlorosalicylic acid, with minor amounts of 2,5-dihydroxy-3,6-dichlorobenzoic acid and 5 hydroxydicamba found. Dicamba is highly mobile in soil, and significant leaching is possible; its water solubility is 6.5 g.L-1 (25 degrees C) and it has a log octanol-water partition coefficient of 0.477. Acute and chronic toxicological studies for all nontarget plants and animals were also reviewed. The major groups of organisms for which toxicological data were collected were freshwater fish, invertebrates and plants, tame hays and cereals, legumes, and other crops, and livestock poultry and mammals. The acute toxicity (< or = 96-hr LC50) to freshwater fish ranged from 28 to 516 mg.L-1, whereas that for invertebrates ranged from 3.9 to > 100 mg.L-1. No chronic data were found for either of these groups. The chronic EC50 to 14 freshwater algae, based on growth inhibition, ranged from 100 to > 10,000 micrograms.L-1. No studies on freshwater macrophytes or any marine organisms were found. Agricultural crops exhibited varying toxicity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234943 TI - Fate and effects of the surfactant sodium dodecyl sulfate. AB - Sodium dodecyl sulfate is the most widely used of the anionic alkyl sulfate surfactants. Its surface-active properties make it important in hundreds of household and industrial cleaners, personal care products, and cosmetics. It is also used in several types of industrial manufacturing processes, as a delivery aid in pharmaceuticals, and in biochemical research involving electrophoresis. SDS synthesis is a relatively simple process involving the sulfation of 1 dodecanol followed by neutralization with a cation source. Purification is accomplished through repeated extraction. It is available commercially in both broad-cut and purified forms. Although its environmental occurrence arises mainly from its presence in complex domestic and industrial effluents, SDS is also directly released in some applications (e.g., oil dispersants and pesticides). Although surfactants are known to significantly contribute to the toxicity of some effluents, no official water quality standards currently exist. Research has shown SDS to be highly biodegradable by a large number of naturally occurring bacteria, and degradation is generally reported to be > or = 90% within 24 hr. The process involves initial enzymatic sulfate liberation and conversion to dodecanoic acid, followed by either beta-oxidative shortening or elongation and desaturation. All surfactant properties are lost after initial sulfate hydrolysis. SDS can enhance absorption of chemicals through skin, gastrointestinal mucosa, and other mucous membranes. Thus, it is used in transepidermal, nasal, and ocular drug delivery systems and to enhance the intestinal absorption of poorly absorbed drugs; enhancement is concentration dependent. Human exposure is mainly through oral ingestion and dermal contact, although cases of respiratory exposure are known. The main sources of daily intake are ingestion of personal care products, residues on insufficiently rinsed utensils, and contaminated drinking water. Uptake, distribution, and excretion of SDS are all rapid. In fish, uptake in various tissues plateaus within 24-72 hr, with elimination occurring within < 24-48 hr; selective accumulation occurs in the hepatopancreas and gall bladder. In mammals, it is readily absorbed via the intestine, colon, and skin. Metabolism is similar in fish and mammals, proceeding from initial omega-oxidation to a carboxylic acid, then to beta-oxidation to butyric acid 4-sulfate, which is finally nonenzymatically desulfurated to gamma butyrolactone and inorganic sulfate. SDS elicits both physical and biochemical effects on cells, with the membrane the primary target structure. Effects are concentration dependent and range from loss of barrier function and increased permeability to complete cell lysis. Hemolysis in mammals is pH dependent.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8234944 TI - Body surface area misconceptions. PMID- 8234945 TI - Key issues in carcinogen risk assessment guidelines, Society for Risk Analysis. PMID- 8234946 TI - An overview of the report: correlation between carcinogenic potency and the maximum tolerated dose: implications for risk assessment. AB - Current practice in carcinogen bioassay calls for exposure of experimental animals at doses up to and including the maximum tolerated dose (MTD). Such studies have been used to compute measures of carcinogenic potency such as the TD50 as well as unit risk factors such as q1 * for predicting low-dose risks. Recent studies have indicated that these measures of carcinogenic potency are highly correlated with the MTD. Carcinogenic potency has also been shown to be correlated with indicators of mutagenicity and toxicity. Correlation of the MTDs for rats and mice implies a corresponding correlation in TD50 values for these two species. The implications of these results for cancer risk assessment are examined in light of the large variation in potency among chemicals known to induce tumors in rodents. PMID- 8234947 TI - The importance of data on mechanism of carcinogenesis in efforts to predict low dose human risk. PMID- 8234948 TI - The value of animal test information in environmental control decisions. AB - Value of information (VOI) analytic techniques are used to evaluate the benefit of performing animal bioassays to provide information about the cancer potency of specific chemical compounds. These tools allow the identification of the conditions in which the cost of reducing uncertainty about potency, by performing a subchronic or chronic bioassay, is justified by the benefit of having improved information for making control decisions. The decision analytic results are readily scaled to apply to a range of human contact rates (exposures) and a variety of control strategies. The sensitivity of results to uncertainty about animal to human extrapolation and the design of the bioassay is explored. An evaluation of the possible gains in general understanding about the mechanisms of carcinogenicity resulting from chronic bioassays is beyond the scope of this approach. PMID- 8234949 TI - Demographic influences on risk perceptions. AB - Over the past 15 years, psychologists have empirically investigated how people perceive technological, consumer, and natural hazards. The psychometric-attitudes to risk being summarized by three factors: "dread," whether the risk is known, and personal exposure to the risk. The results have been used to suggest that certain types of hazards are viewed very differently from other hazards. The purpose of this paper is somewhat different, in that it investigates whether individual demographic characteristics influence psychometric perceptions of risk. This paper makes use of a large, professionally conducted, survey of a wide cross-section of the residents of metropolitan Chicago. One thousand adults were interviewed in a random-digit dial telephone survey, producing a useable dataset of about 800. Data on the three risk factors mentioned above were obtained on 7 point scales for four common hazards: aviation accidents, fires in the home, automobile accidents, and stomach cancer. The survey also collected demographic data on respondents' age, schooling, income, sex, and race. Regressions were then conducted to relate the demographic characteristics to risk perceptions. Some strong general conclusions can be drawn. The results suggest that women, people with lower levels of schooling and income, younger people, and blacks have more dread of hazards. The exception being age-related illnesses which, not unnaturally, are feared by older people. Unlike previous literature, we cannot substantiate the argument that these groups of people are less informed about hazards and thus less accepting of them. The most likely leading explanation of the relationship between demographic factors and dread of a hazard is the perceived personal exposure to the hazard.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8234950 TI - Risk perception among nuclear power plant personnel: a survey. AB - This study investigated risk perception, well-being, and organizational commitment among nuclear power plant personnel. The study group, 428 employees from a nuclear power plant, completed a questionnaire which included the same questions as those in previous surveys on risk perception of lay persons and industrial workers. Hazards at work were not seen as a sizable problem by nuclear power plant personnel. The study group estimated the safety of nuclear power plants better and the possibility of serious nuclear accident as more unlikely than the general public. Compared to employees in other industrial companies, the overall perceived risks at work among plant personnel did not exceed the respective perceptions of the reference groups. Risk-related attitudes did not explain well-being among plant personnel, but the relationship between the perceived probability of a serious nuclear accident at work and organizational commitment yielded to a significant correlation: Those plant workers who estimated the likelihood of an accident higher were less committed to the organization. PMID- 8234951 TI - Politics and scientific expertise: scientists, risk perception, and nuclear waste policy. AB - To study the homogeneity and influences on scientists' perspectives of environmental risks, we have examined similarities and differences in risk perceptions, particularly regarding nuclear wastes, and policy preferences among 1011 scientists and engineers. We found significant differences (p < 0.05) in the patterns of beliefs among scientists from different fields of research. In contrast to physicists, chemists, and engineers, life scientists tend to: (a) perceive the greatest risks from nuclear energy and nuclear waste management; (b) perceive higher levels of overall environmental risk; (c) strongly oppose imposing risks on unconsenting individuals; and (d) prefer stronger requirements for environmental management. On some issues related to priorities among public problems and calls for government action, there are significant variations among life scientists or physical scientists. We also found that--independently of field of research--perceptions of risk and its correlates are significantly associated with the type of institution in which the scientist is employed. Scientists in universities or state and local governments tend to see the risks of nuclear energy and wastes as greater than scientists who work as business consultants, for federal organizations, or for private research laboratories. Significant differences also are found in priority given to environmental risks, the perceived proximity of environmental disaster, willingness to impose risks on an unconsenting population, and the necessity of accepting risks and sacrifices. PMID- 8234952 TI - Radon testing behavior in a sample of individuals with high home radon screening measurements. AB - Although radon exposure has been identified as the second leading cause of lung cancer, fewer than 6% of U.S. homeowners test their homes for radon. This report examines participants' follow-up radon testing behavior subsequent to receiving an initial screening radon level greater than 20 pCi/L. Sixty-two participants in the Iowa State-Wide Rural Radon Screening Survey who had radon screening measurements over 20 pCi/L were questioned by phone survey 3 months after receipt of their radon screening result to assess: whether participants were aware of radon's health risk; if participants recalled the radon screening results; how participants perceived the relative health risk of radon and whether participants planned follow-up radon testing. Only 19% of the respondents specifically identified lung cancer as the possible adverse health outcome of high radon exposure, and the majority of participants underestimated the health risks high radon levels pose when compared to cigarettes and x-rays. In addition, less than one third (29%) of the participants actually remembered their radon screening level within 10 pCi/L 3 months after receiving their screening results. Only 53% of the individuals correctly interpreted their screening radon level as being in the high range, and only 39% of the participants planned follow-up radon measurements. Receipt of radon screening test results indicating high radon levels was not an adequate motivational factor in itself to stimulate further radon assessment or mitigation. Our findings suggest that free radon screening will not result in a dramatic increase in subsequent homeowner initiated remediation or further recommended radon testing. PMID- 8234953 TI - Perceptions of nuclear and other risks in Japan and the United States. AB - As part of a study of nuclear power development in Japan and the United States, surveys of perceptions of risk toward 30 activities, substances, and technologies have been carried out in the Pacific Northwest and Tokyo, Japan. The results show that people in both countries have the highest level of dread toward nuclear waste disposal, nuclear accidents, and nuclear war, greater even than their dread of crime and AIDS. In addition to comparisons of dread, the paper also discusses similarities and differences between Japanese and American responses for other dimensions of risk perception. PMID- 8234954 TI - Baseline risk and preference for reductions in risk-to-life. AB - The typical model of individual attitudes toward risk-to-life suggests that an individual's willingness to pay for a reduction in mortality risk increases with the baseline risk. The higher-baseline hypothesis has been the subject of several empirical tests but results have so far been mixed. Using survey evidence, we present a situation in which subjects do prefer to reduce risks for which the baseline is higher. This finding is robust to several alternative explanations. Survey responses reflect subjects' concerns about government effectiveness in risk reduction, environmental effects associated with the various hazards, and other idiosyncratic elements of the risks; however, these concerns appear to occur in addition to, not in lieu of, the preference to reduce higher risks. PMID- 8234955 TI - An enforceable indoor air quality standard for environmental tobacco smoke in the workplace. AB - Environmental tobacco smoke (ETS) has recently been determined by U.S. environmental and occupational health authorities to be a human carcinogen. We develop a model which permits using atmospheric nicotine measurements to estimate nonsmokers' ETS lung cancer risks in individual workplaces for the first time. We estimate that during the 1980s, the U.S. nonsmoking adult population's median nicotine lung exposure (homes and workplaces combined) was 143 micrograms (micrograms) of nicotine daily, and that most-exposed adult nonsmokers inhaled 1430 micrograms/day. These exposure estimates are validated by pharmacokinetic modeling which yields the corresponding steady-state dose of the nicotine metabolite, cotinine. For U.S. adult nonsmokers of working age, we estimate median cotinine values of about 1.0 nanogram per milliliter (ng/ml) in plasma, and 6.2 ng/ml in urine; for most-exposed nonsmokers, we estimate cotinine concentrations of about 10 ng/ml in plasma and 62 ng/ml in urine. These values are consistent to within 15% of the cotinine values observed in contemporaneous clinical epidemiological studies. Corresponding median risk from ETS exposure in U.S. nonsmokers during the 1980s is estimated at about two lung cancer deaths (LCDs) per 1000 at risk, and for most-exposed nonsmokers, about two LCDs per 100. Risks abroad appear similar. Modeling of the lung cancer mortality risk from passive smoking suggests that de minimis [i.e., "acceptable" (10(-6))], risk occurs at an 8-hr time-weighted-average exposure concentration of 7.5 nanograms of ETS nicotine per cubic meter of workplace air for a working lifetime of 40 years. This model is based upon a linear exposure-response relationship validated by physical, clinical, and epidemiological data. From available data, it appears that workplaces without effective smoking policies considerably exceed this de minimis risk standard. For a substantial fraction of the 59 million nonsmoking workers in the U.S., current workplace exposure to ETS also appears to pose risks exceeding the de manifestis risk level above which carcinogens are strictly regulated by the federal government. PMID- 8234956 TI - Using decelerative contingencies to reduce the self-injurious behavior of people with multiple handicaps: the effects of response satiation? AB - The response satiation model of instrumental performance was used to establish contingencies to reduce the self-injurious behavior (SIB) of two adults with profound mental retardation and multiple handicaps. In Experiment 1, withdrawal designs indicated behavior reductions of 68% and 82%, respectively, when contingencies were implemented that required the participants to engage in leisure activities at greater than baseline levels if they continued to perform the SIB at their baseline level. Experiment 2 involved the transfer of the treatment to the participants' respective living areas using staff as trainers. The largest reductions in behavior were associated with treatment packages that contained response satiation contingencies, and generality of the treatment packages across locations and trainers occurred. The limitations of this study as an analysis of response satiation theory were noted; however, it was concluded that the response satiation approach to establishing contingencies for reducing SIB was worthy of future investigation. PMID- 8234957 TI - The effects of aerobic exercise on psychological and behavioral variables of individuals with developmental disabilities: a critical review. AB - Physical fitness of persons who are developmentally disabled has received relatively little attention in the special education literature when compared to intellectual functioning (e.g., learning, memory, and language) and to acquisition of functional skills (e.g., self-care, community, and vocational). Despite an increased interest in recreational programming stimulated by the concept of functional curricula, teachers may still be reluctant to include physical fitness activities in their students' schedules. Perhaps physical fitness programming for those with developmental disabilities would have wider appeal and application if it were embedded in the broader context of psychological and behavioral change (i.e., engagement in exercise produces generalized changes beyond direct improvement in physical well-being). This article is a review and critique of literature that focuses on the effects of participation in aerobic exercise on three classes of psychological/behavioral variables for persons with mental retardation and associated disabilities. The methodology that characterizes this literature is analyzed, and recommendations for future research are proposed. PMID- 8234958 TI - Effectiveness of home-based early intervention on the language development of children of mothers with mental retardation. AB - The authors evaluated the effects of a home-based parent training program for mothers with mental retardation on the language development of their children who were at risk for language delay. The participants, 28 mothers labelled mentally retarded with children under 28 months of age, initially showed significantly fewer positive mother-child interactions and child vocalizations and verbalizations than did a comparison group of 38 families with children of similar age whose mothers were not mentally retarded. The 28 mothers with low IQ were then matched on child entry age and randomly assigned to either an interaction training or attention-control group (this group received training in safety and emergency skills unrelated to interactions). Interaction training consisted of verbal instruction, modelling, feedback, and tangible reinforcement. After training, the training group scores were no longer lower than those of the comparison group of mothers without mental retardation and were also significantly higher than the scores of the attention-control group on all maternal positive interactions, child vocalizations, verbalizations, and language and social domains of the Bayley Scales of Infant Development. Speech emerged significantly sooner in the training group as compared to the control group. The training group parents and children maintained improvements up to 82 weeks following training, and the attention-control group, when subsequently trained, replicated the original training group results. Thus, home-based parent training increased positive maternal interactions of mothers with mental retardation, which facilitated language development in their young children. PMID- 8234959 TI - Comprehension monitoring skills of adults with mental retardation. AB - The ability of 30 adults with mild and moderate levels of mental retardation to monitor their comprehension while performing a direction-following task was examined. Subjects were "employed" to aid the investigator in compiling objects for gift bags. The task consisted of 60 directions (e.g., "Give me a blue pencil"), of which 6 contained trouble sources (ambiguous directions, unintelligible words, and compliance problems). Each subject's response to the directions involving trouble sources was scored to determine if the subject (a) demonstrated effective comprehension monitoring (as indicated by an immediate awareness of the problem and effective attempt to rectify the problem), (b) requested clarification after attempting to comply with the direction and being unable to do so, (c) demonstrated ineffective comprehension monitoring (as indicated by an awareness of the problem but an ineffective means of dealing with it), or (d) showed no awareness of the trouble source. The ability of subjects to monitor comprehension varied with type of trouble source. Ambiguous trouble sources were the most difficult for the subjects to detect, and compliance problem trouble sources were the most frequently identified. PMID- 8234960 TI - New approaches in asthma and COPD. October 1992, Davos, Switzerland. Proceedings. PMID- 8234961 TI - Immunohistology in bronchial asthma. PMID- 8234962 TI - The clinical relevance of bronchial biopsies in asthma. PMID- 8234963 TI - Practical aspects of bronchoalveolar lavage. PMID- 8234964 TI - Bronchial hyperresponsiveness today. PMID- 8234965 TI - Inflammation and bronchial hyperresponsiveness in allergic asthma and chronic obstructive pulmonary disease. PMID- 8234966 TI - Direct and indirect bronchial responsiveness. PMID- 8234967 TI - Epidemiology and the concept of underlying mechanisms of nocturnal asthma. AB - Nocturnal symptoms are common in asthma, even when patients are regularly seen at an outpatient clinic. Inflammation is generally accepted as a general feature of asthma and the severity of this basic inflammatory process can be increased by exogenous triggers such as exposure to allergens and nonallergic stimuli. Superimposed endogenous circadian rhythms may play a more important and intricate role in the circadian modulation of the inflammatory process by changing the number of cells, their release of mediators and/or the susceptibility of airway smooth muscle and vasculature. For instance, an increase in vagal tone may induce nocturnal bronchoconstriction which is further enhanced by falling catecholamine levels. Together, the reduced nocturnal catecholamine levels and the diminished bronchodilating capacity of the NANC system and the low cortisol levels oppose possible protection against inflammatory processes leading to nocturnal airflow obstruction. PMID- 8234968 TI - Pulmonary rehabilitation in asthma and COPD, physiological basics. PMID- 8234969 TI - Prevalence and pathophysiology of nutritional depletion in chronic obstructive pulmonary disease. PMID- 8234970 TI - Chronic obstructive lung disease and respiratory muscle function: the role of nutrition and exercise training in cystic fibrosis. AB - Malnutrition may be an extremely important complicating factor in patients with a variety of chronic lung diseases. The impact of malnutrition on lung disease and respiratory muscle function has been extensively studied in patients with chronic lung disease due to cystic fibrosis (CF). In patients with CF it appears that malnutrition and deterioration of lung function are closely interrelated and interdependent, with each affecting the other, leading to a spiral decline in both. The occurrence of malnutrition during childhood seems to be associated with impaired growth and repair of the airway walls. Therefore, in children, when growth in body-length may be expected, prevention of malnutrition is associated with better lung function and improved survival. Nutritional intervention in adults, when no further growth may be expected, will only lead to an increase of body fat. However, when combined with a physical training programme it may lead to an improvement in bodyweight, respiratory muscle function, lung function and exercise tolerance. In addition, this approach will lead to an increase in both respiratory and other muscle mass. PMID- 8234971 TI - Methotrexate in the treatment of severe asthma. PMID- 8234972 TI - Lung transplantation and chronic obstructive pulmonary disease. PMID- 8234973 TI - Coping with asthma. AB - Coping is a dynamic process playing a major role in a person's behaviour to respond in the best possible way to his environment. Behaviour and environment mutually influence one another. Non-compliance appears in all therapies prescribing the patient to administer his own medication. A number of studies shows that approximately 50% of the chronic patients do not take their medication or do not take it in accordance with the prescription. Knowledge and skills are important variables for maintaining desired behaviour of asthma patients. In order to cope adequately with the disease it requires the patient's understanding of the daily use of an often complex medication regimen. With respect to asthma patients health education is particularly aimed at supplying them with knowledge and skills that enable them to cope adequately with the disease in everyday life. Moreover, health education functions to motivate patients. On investigation into the relation between coping and disease it is important that one should not only focus on the disease and the treatment, but particularly on the relation between the patient, his immediate environment and society, which relation should be adapted. PMID- 8234974 TI - Bronchial epithelial shedding in asthmatics and non-asthmatics. PMID- 8234975 TI - [Norwegian scabies in AIDS]. PMID- 8234976 TI - [Epidemic outbreak of scabies related to a patient with human immunodeficiency virus infection and Norwegian scabies]. AB - A patient with AIDS was admitted to our hospital because of pruritic and disabling erythroderma, it was treated with Lindane without improvement. Skin biopsy of a crusty lesion showed Sarcoptes scabiei hominis, and the diagnosis of Norwegian scabies was made. After a long and intensive treatment with several drugs, cure was achieved. In the following three months we detected 42 people with scabies, all in some relation with the index case. This feature of high contagiousness of Norwegian scabies in a constant, through the scanty literature we have found; and it is due to its diagnostic and therapeutic problems. PMID- 8234977 TI - [Epidemiologic and clinical characteristics and clinical course of the HIV positive patient infected by heterosexual transmission]. AB - We reviewed the features of patients with human immunodeficiency virus (HIV) infection, whose only way of acquisition was heterosexual contacts, analyzing differences with others risk groups. Epidemiological features (age, gender, family situation, socioeconomic status and sexual behavior), clinical manifestations immunological status, and evolution of 40 patients with HIV infection through heterosexual contact are studied, and compared with others risk groups. All were attended in our center from 1985 to October 1991; 15 were in stage IV. Along the study period, the proportion of heterosexual patients has steadily increased to represent 40% on total number of acquired immunodeficiency syndrome (AIDS) cases diagnosed in 1991. This study allow to establish two groups of patients depending on the gender. All men had had occasional sexual contacts with prostitutes, most had no stable partner, they mainly were between their fourth and sixth decade of life, and they were stable partners of seropositive patients or prostitutes, they had an averaged age significantly lower than men, and they were diagnosed in earlier stages. Two groups with different features can be established among the patients infected through heterosexual contacts. Men are adults, and acquired HIV infection through contacts with prostitute; they are diagnosed in late phases of the disease. On the contrary, women are younger, with no homogeneous patterns of behavior and they are diagnosed in earlier stages. Complications throughout the evaluation were similar in the heterosexual and in the other groups. PMID- 8234978 TI - [Anatomopathologic evaluation of the pathergic reaction in Andalusian patients with Behcet's disease]. AB - Pathergic reaction (PR) has been recently included in a system of diagnostic criteria for Behcet's disease (BD), justified for its high frequency in several countries like Turkey (58%). As in occidental countries it is much less common, we advise to evaluate its diagnostic usefulness in our country. An anatomopathological study of the PR in 23 patients with BD, and 14 healthy volunteers, was conducted. Punch samples were stained with hematoxylin-eosine and for immunofluorescence. Macroscopically, there was no PR in any person (neither patients nor controls). About half of the patients showed leukocytoclastic vasculitis and/or cellular infiltration (mainly neutrophilic). These findings are no specifics (they were seen in same controls) and have a low diagnostic sensitivity. Authors consider unsuitable the inclusion of PR, as an independent BD diagnostic criterion, in our country. PMID- 8234979 TI - [Pheochromocytoma with ectopic secretion of ACTH]. AB - Pheochromocytoma is a tumor displaying a considerable degree of clinical polymorphism both with respect to its location and the secretion of different types of catecholamines and its capacity to synthesize several ectopic hormones. The present work reports on a case of pheochromocytoma with secretion of ACTH that exhibited several clinical and biological peculiarities. Firstly, the clinical picture of the patient was characterized by a predominance of manifestations of adrenal hyperfunction as compared with those of the original tumor. Secondly a striking finding was an important degree of neutrophilia without band forms and a marked hypokalemia produced by the joint secretion of catecholamines and steroid hormones, as demonstrated later. Finally, the images obtained with computerized tomography were very characteristic, such that together with the above data it was possible to establish a preoperative diagnosis. PMID- 8234980 TI - [Type I familial pseudohypoparathyroidism: clinical and hormonal study]. AB - Pseudohypoparathyroidism is a rare disease characterized by a target tissue resistance to the action of the parathyroid hormone (PTH). Frequently, others hormonal disorders can be associated. We report a family with pseudohypoparathyroidism type I, and discuss the clinical-hormonal alterations founded; it is emphasized that while there were no phenotypical differences among the family members; from the hormonal features point of view, some differences do exist, and several degrees of peripheral resistance were noted. PMID- 8234981 TI - [Functional abnormality of the diaphragm in the "shrinking lung" in systemic lupus erythematosus]. AB - Functional respiratory disorders in patients with systemic lupus erythematosus (SLE) have been previously reviewed. We report a patient with SLE who developed progressive dyspnea with a restrictive pattern in the functional respiratory tests, and diaphragmatic disfunction. Symptoms and functional tests improved after a treatment with steroids. We also review the pathophysiologic mechanisms of this disorder. PMID- 8234982 TI - [Cognitive functions in agenesis of the corpus callosum]. AB - Four patients with agenesis of the corpus callosum diagnosed through neuroradiologic tests has been studied, from the upper cerebral functions point of view. Three had consulted because of seizure, and the other because of cerebellar hemorrhage. Multiple fields of the cognitive functions were examined and the presence of an interhemispheric disconnection syndrome was investigated. All tests were absolutely normal in the four patients. We discuss clinical and radiologic aspects as well as other associated malformations. PMID- 8234983 TI - [Myopathy and polyneuropathy caused by colchicine]. AB - A forty-six year old man with chronic renal failure and a toxic chronic liver disease developed progressive muscle weakness after a long trial with colchicine. Physical exam revealed muscle weakness and proximal muscle atrophy with hyporeflexia. Serum levels of creatine kinase were high and signs of myopathy and axonal and demyelinating polyneuropathy was evidenced in electrophysiological studies. Muscle biopsy disclosed a vacuolar myopathy, disruption of myofibers and dilatation of sarcoplasmic reticulum. The clinical pictures was attributed to a toxic myopathy and polyneuropathy due to colchicine, thus this treatment was discontinued. Four week later, the patient was symptom free, the levels of seric creatine kinase were normal and a new muscle biopsy was normal, with disappearance of previous histological findings. PMID- 8234984 TI - [Frontal tumor and mental deterioration]. PMID- 8234985 TI - [Massive hemoptysis in a patient with arterial hypertension]. PMID- 8234986 TI - [Mortality due to respiratory diseases in Spain (1977-1985)]. AB - A descriptive and comparative epidemiological is made of mortality due to respiratory disease by provinces in Spain. The study period covers 1977 to 1985, this being the last year reported by the Natural Movement of the Spanish Population (Movimiento Natural de la Poblacion Espanola). In particular, 5 causes of death were analyzed in accordance to the International Classification of Disease (IX Revision): respiratory tuberculosis, influenza, chronic pulmonary disease, pneumonia and other pathologies of the respiratory apparatus. The results reveal an annual increase in these diseases. Standardized mortality ratio in turn reflects the provinces with increased mortality, the distribution of which varies for each pathology studied as a result of the different factors involved. Variation is also seen in terms of sex. Thus, influenza predominates among women, whereas males are more frequent as regards the remaining pathologies. Mortality due to pneumonia is similar in both sexes, this being the disease exhibiting the greatest increase during the study period. The results obtained may be of use in protocolizing resources for better control and prevention of respiratory disease in this country. PMID- 8234987 TI - [Immunopathogenesis of chronic autoimmune hepatitis]. PMID- 8234988 TI - [A male with human immunodeficiency virus infection and pulmonary nodule]. PMID- 8234989 TI - [Lumbago, infrequent presentation form of neuroblastoma in the adult]. PMID- 8234990 TI - [Erythrocyte dehydrogenase glucose-6-phosphate deficiency and selective IgA deficiency: a familial study]. PMID- 8234991 TI - [Spontaneous peritonitis caused by Campylobacter jejuni]. PMID- 8234992 TI - [Primary thyroid angiosarcoma]. PMID- 8234993 TI - [Hypocholesterolemia and hypoapolipoproteinemia A1 in a female patient diagnosed with acute lymphoblastic leukemia]. PMID- 8234994 TI - [The concept of left ventricular hypertrophy: the frontiers between physiology and pathology]. PMID- 8234995 TI - [Cardiovascular risk factors in a population of schoolchildren in Asturias]. AB - A transversal epidemiological study was carried out to discover the most important cardiovascular risk factors in the child population of Asturias, Spain. The study took place over a period of 4 years (1987-91) and consisted of a representative sample of boys and girls (6-13 years of age) from different socioeconomic and cultural backgrounds. Biochemical factors (serum total cholesterol, HDL-cholesterol, serum triglycerides and glucose) and anthropometric measures to evaluate obesity were analysed. Hypercholesterolemia appeared to be the most prevalent risk factor with a prevalence of 38.27% and a mean value of 178.31 mg/dl, followed by obesity (15.58%), hypertriglyceridemia (14.05%) and hyperglycemia (0.77%). Risk factors were more frequent in girls than boys, and in younger children in relation to older ones. Hypercholesterolemia and obesity were more prevalent in children from an urban environment while hypertriglyceridemia was the main risk factor in those from a rural one. Considering the high prevalence of the evaluated risk factors, the prevention and control of such is of extreme importance in leading children to becoming healthy adults. PMID- 8234996 TI - [Cholesterol and other risk factors in schoolchildren]. PMID- 8234997 TI - [The prevalence of cardiovascular risk factors in a population of essential hypertension patients]. AB - INTRODUCTION AND OBJECTIVES: High blood pressure is a cardiovascular risk factor whose coincidence with other risk factors considerably increases the probability for coronary heart disease. This study investigates the prevalence of different risk factors in hypertensive subjects. METHODS: We studied 891 patients with high blood pressure in their first visit to a hospital hypertension unit between 1987 and 1991. We assessed their clinical evolution and the prevalence of obesity, smoking, alcohol consumption, elevated blood pressure, diabetes, sedentary lifestyle, left ventricular hypertrophy and family history of high blood pressure. RESULTS: Sixteen per cent of the hypertensive subjects related previous history of evolutive accident. Stroke was the most frequent one. Prevalences of cardiovascular risk factors were as follows: family history of high blood pressure 53%, sedentary lifestyle 52%, elevated blood cholesterol 37%, smoking 35%, obesity 33%, left ventricular hypertrophy 16%, alcohol consumption 13% and diabetes 11%. Obesity prevalence was twice as high in women than in men. Males had higher prevalence for left ventricular hypertrophy, smoking and alcohol consumption. Diabetes was more prevalent in hypertensive patients older than 50 years. Prevalence of high blood cholesterol was greater in the group of women older than 50 years. CONCLUSIONS: Hypertensive patients have high prevalences of other cardiovascular risk factors. Their treatment should be aimed to improving the individual profile of cardiovascular risk. PMID- 8234998 TI - [The acute and chronic effects of 10-mg nitroglycerin patches in stable angina of effort]. AB - INTRODUCTION AND OBJECTIVES: To assess the anti-ischemic efficacy of nitroglycerin patches (10 mg/day), we studied, by means of serial exercise testing (Bruce protocol), 10 patients with stable effort angina in a randomized, placebo-controlled, cross-over, double-blind essay. METHODS: Patients were exercised 1, 4, 12 and 24 hours after a single patch, and 4 and 12 hours after a 48 hours therapy course. Chronic therapy was assessed after both continuous and intermittent (intermission of 12 hours) patch application. RESULTS: After single patch, time to angina and time to 1 mm ST depression were significantly increased with respect to placebo at 1-hour test (83 +/- 27 s and 119 +/- 39 s, respectively), 4-hour test (100 +/- 34 s and 87 +/- 29 s, respectively) and 12 hour test (46 +/- 15 s and 64 +/- 20 s, respectively). No effect was demonstrated at 24-hour test. After continuous treatment no differences with respect to placebo were found at any test. After intermittent treatment time to angina was prolonged (75 +/- 23 s) only at 4-hour test, and time to 1 mm ST depression at 4 hour test (61 +/- 19 s) and 12-hour test (41 +/- 14 s). CONCLUSIONS: Nitroglycerin patches improve parameters of exercise ischemia for a 12 hours period. Tolerance is developed very early and provokes absolute lack of efficacy. Tolerance can be avoided with intermittent patch application. No treatment schedule shows 24 hours efficacy. PMID- 8234999 TI - [Does captopril possess antianginal actions? An evaluation of its effects on myocardial ischemia by stress tests in patients with stable angina and normal ventricular function]. AB - INTRODUCTION: We have studied the effects of captopril on myocardial ischaemia in normotensive and hypertensive patients with coronary artery disease and stable effort-induced angina. STUDY DESIGN: A maximal treadmill effort test (Bruce modified) was obtained before and after 60 min administration of 25 mg. p.o. of captopril. In a first open pilot essay, 12 patients were studied. In a double blind randomized, placebo-controlled, without crossover, 20 patients with the same characteristics and methodology were divided in two groups (10 in placebo and 10 in captopril group) and compared in a maximal treadmill effort test. RESULTS: In the pilot essay, the second test don't showed increments in heart rate, systolic and diastolic blood pressure showed a significant reduction at rest (p < 0.01) and at 1 mV ST segment depression (p < 0.01). Captopril increased exercise duration at 1 mV ST segment depression, time to angina and total exercise time (p < 0.01). In double-blind essay, captopril group results were similar to the pilot essay. In comparing results between placebo and captopril groups the most relevant differences were: significant increment in the time to 0.1 mV ST segment depression (p < 0.01), to angor (p < 0.05) and total exercise duration (p < 0.01). Maximal work-load sustained increased significantly with captopril (7.43 +/- 2.1 to 10.34 +/- 1.8 METS) (p < 0.01). CONCLUSIONS: We conclude that captopril used in monotherapy in patients with chronic stable angina, seems to reduce clinical and electric ischemia and to ameliorate maximal exercise duration and work-load. PMID- 8235000 TI - [Cardiac arrhythmias (X). Vasovagal syncope. Its physiology, diagnosis and treatment]. PMID- 8235001 TI - [Revascularization in multivessel disease: the viewpoint of the clinician]. PMID- 8235002 TI - [Revascularization in multivessel disease: the viewpoint of the surgeon]. PMID- 8235003 TI - [Revascularization in multivessel disease: the viewpoint of the interventionist cardiologist]. PMID- 8235004 TI - [The adult respiratory distress syndrome as a complication of the APSAC treatment of acute myocardial infarct. Apropos a case]. AB - Thrombolytic therapy has shown to be effective in acute myocardial infarction, and its benefits on left ventricular function and later survival are well known. However it is not a therapy free of complications and side effects. Allergic reactions, anafilaxia, hypotension, and several kinds of hemorrhages have been reported. Adult respiratory distress syndrome after streptokinase administration has been also described, and one case, recently communicated, after APSAC therapy. We present the case of a male with acute myocardial infarction who developed adult respiratory distress syndrome after APSAC therapy, with different outcome than the first case published in the literature. Finally, we discussed the mechanisms by means these drugs can produce such a complication. PMID- 8235005 TI - [A large aneurysm of the interatrial septum simulating an atrial mass. Its identification by transesophageal echocardiography]. AB - A patient with a large atrial septal aneurysm mimicking a right atrial mass is reported. Diagnosis was correctly obtained by transesophageal echocardiography. Clinical importance of this entity and usefulness of transesophageal echocardiography in its diagnosis is discussed. PMID- 8235006 TI - [Acute pericarditis as the initial presentation of a primary cardiac angiosarcoma. The anatomicoclinical correlations]. AB - We report the clinic and postmortem pathologic examination of a 38-years-old man with a primary angiosarcoma of right atrium and many widespread metastatic. Both his presentation, a clinical course of acute pericarditis self-limited, as the negatives complementary examination, give at case unusual characteristics, for his high malignancy. PMID- 8235007 TI - [Differences in prediction of mortality and non-fatal complications after acute myocardial infarction: a multivariate analysis]. AB - The global prognostic after acute myocardial infarction depends of the left ventricular function and the residual ischemia. However, there is controversy about the capacity of some variables reflecting this prognostic factors to predict futures complications. The main objective of this study is to know the influence of the kind of complications in the prognostic value of the variables more frequently used in the clinical practice. We studied 121 consecutive patients with acute myocardial infarction discharged from hospital. The previous and during acute infarction clinical variables were analyzed. The regional and global left ventricular function was studied by 2D echocardiography. An exercise test symptoms-limited was performed at 3rd week after discharge in 94 patients selected. The patients were followed during selected one year. The first year cardiac mortality rate was 8.6%. In the multivariate, the independent predictors of mortality were: age > 60 years (p = 0.02) and the impossibility to perform exercise test (p = 0.0002). In the univariate analysis the echocardiographic motility score > or = 17, eyection fraction < 35% and Killip class > or = II were also death predictors. Non fatal complications (angina, reinfarction o heart failure) were present in 47 patients (42.3%) of 111 survivors. The ischemic complications were predicted in multivariate analysis only by postinfarction angina (p = 0.0007), and heart failure by eyection fraction < 35% (p = 0.006), previous infarct (p = 0.002) and Killip class (p = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235008 TI - [Post-infarction prognosis: separating ischemia from heart failure and death]. PMID- 8235009 TI - [Evaluation of home physical training soon after acute myocardial infarction]. AB - Little attention has been directed to the utility of unsupervised physical training at home in postmyocardial infarction patients. The present investigation was designed to assess the effects of exercise training at home on functional parameters soon after myocardial infarction. One hundred and eighteen male postmyocardial infarction patients under 70 years old, that performed a predischarge exercise test, were randomly allocated soon after the acute event, into a 12-14 weeks unsupervised physical training at home and into a control group. The control group patients were informally encouraged to practice exercise. Before training, the cardiorespiratory parameters were similar in both groups. After training, improvement of all these variables was seen in both groups, although the magnitude was greater in the training group. Maximal oxygen consumption increased 7.6 +/- 3.6 ml/kg/min in the training group by 3.8 +/- 2.6 ml/kg/min in the control group (p = 0.00001), maximal heart rate increased 14.6 +/- 13.4 by 8.4 +/- 5.7 beats/min (p = 0.001), and similar results were obtained with maximal double product and exercise test duration (33 +/- 46 versus 17 +/- 12, p = 0.009; 2.2 +/- 1.1 versus 0.9 +/- 1.1 min, p = 0.00001) always in trained patients favour. Nine trained patients and 16 control patients showed myocardial ischemia in both exercise tests. At ischemic threshold, the achieved improvement was again greater in trained patients, concerning to heart rate (p = 0.02), double product (p = 0.0005) and exercise time (p = 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235010 TI - [Validation of a new questionnaire to evaluate the quality of life in patients after myocardial infarction]. AB - Quality of life is an important measurement in chronic diseases and especially in patients after myocardial infarction. We designed and structured a new Questionnaire of Quality of Life for Spanish speaking patients from the existing English questionnaires. The new Questionnaire of Velasco-Del Barrio has a total of 44 items, grouped in 9 subscales: health, sleep and rest, emotional behaviour, concerns to the future, mobility, social relations alertness behaviour, communication, and work and leisure time. The questionnaire has been validated taking as a gold standard, the Quality of Life Questionnaire for Myocardial Infarction (QLMI-Q) of Oldridge. The reliability and the reproducibility have also been studied. The validity of the Velasco-Del Barrio Questionnaire was high (r = 0.81), as its reproducibility (0.75) and reliability (0.90). We assume that the usefulness of the new Questionnaire is at least, similar to that of the Oldridge's one, and that it can be recommended in postinfarction patients, even though it can be more advantageous in patients of Spanish speaking cultures. PMID- 8235011 TI - [Probability analysis of the value of ergometry in the detection of restenosis after coronary angioplasty]. AB - To evaluate the usefulness of treadmill exercise test in the diagnosis of coronary restenosis, we have chosen a sequential and probabilistic analysis. The rate of coronary restenosis found in the sample (n = 213) was of 30%. This initial probability of having restenosis was radically changed by the recurrence of typical angina: 77% in patients with this symptom, 17% in those without (p < 0.001). In contrast, the qualitative result of the exercise test was unable to change significantly the probabilities established after anamnesis. This was due to a lower specificity for exercise test (76%) than for angina (93%), the sensibility being the same (56%) for both. Employment of a discriminant value that integrates different ergometric variables did not enhanced sensibility, but allowed the identification of a good number of false positive results and therefore palliated the negative impact of faulty specificity. Being applied to a "discordant group" (no angina/positive exercise test, n = 34) was able to achieve a significant change in probability: from 15% to 50% or 4% (p < 0.01), according to the discriminant being lesser or greater than a preestablished value. In conclusion, prevalence of the event and evaluation of symptomatology are both important to estimate the probability of suffering coronary restenosis. On the contrary, exercise test, in the absence of a weighty method, adds more confusion than clarity to the analysis of the problem. Our results can be used to improve the follow-up of patients who have been successfully dilated by angioplasty. PMID- 8235012 TI - [Stable single-vessel disease: revascularization or medical treatment? Point of view of the intervention cardiologist]. PMID- 8235013 TI - [Stable single vessel disease: revascularization or medical treatment? Point of view of the cardiac surgeon]. PMID- 8235014 TI - [Stable single vessel coronary disease: revascularization or medical treatment? Point of view of the clinical cardiologist]. PMID- 8235015 TI - [Cardiac arrhythmia (IX). Indications and methods of cardiac stimulation in the 90s]. PMID- 8235016 TI - [Brucella infective endocarditis on sclerodermic mitral valve. Report of a case]. AB - We report the case of a 67 years old woman diagnosed of CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysfunction, sclerodactyly, telangiectasias) variant of generalized scleroderma, that suffered a Brucella endocarditis on mitral valve affected by the primary disease. We examine the cardiac lesions in progressive systemic sclerosis, pointing up the rare involvement of the valves, together with the evolution and treatment of Brucella endocarditis, uncommon entity. PMID- 8235017 TI - [Left coronary artery-right ventricle fistula secondary to heart surgery. Non invasive diagnosis with Doppler-color bidimensional echocardiography]. AB - A case of 20 year-old man with a left coronary artery fistula to right ventricle secondary to open heart surgery is reported; he was asymptomatic and a continuous precordial murmur was found on physical examination. Two dimensional echocardiography-Doppler color flow imaging visualized a dilated left coronary artery with flow into the right ventricle. The discovery was confirmed by coronary angiography. We conclude that two dimensional echocardiography Doppler color flow imaging may be a method included in evaluation of pediatric patients with coronary artery fistula suspicion. PMID- 8235018 TI - [Aortic hypoplasia in a case of single ventricle with normally related vessels]. AB - In cases with double inlet left ventricle when there exists a hypoplasia of one of the great arteries, the affected vessel arises from the accessory right ventricular chamber. This is due in general to the presence of a restrictive bulbo-ventricular foramen that results in hypoplasic vessel. The case we present in this paper has unusual anatomic morphology: the hypoplasic vessel (the aorta) arises from the main ventricular chamber. We analyse the electrocardiographic findings useful for the differential diagnosis from cases of isolated aortic coarctation of hypoplasia. We review also the intraventricular anatomic malformations that may cause the presence of a hypoplasic vessel. Even if, our patient died in the neonatal period, the adequate surgical procedure for this case is reviewed. PMID- 8235019 TI - [The role of endoscopic intervention in the treatment of bronchial cancer]. PMID- 8235020 TI - [Role of chemotherapy in the treatment of advanced stage non-small-cell bronchial cancer]. PMID- 8235021 TI - [Pulmonary emphysema: quantification using computed tomography and correlations with respiratory function tests]. AB - Pulmonary emphysema can be defined in precise anatomical terms. Only histological examination of an entire lung will enable the extension and the severity of the emphysema to be fully assessed. The authors propose a visual score, using computed tomography to quantify emphysema in 61 subjects. 51 had chronic airflow obstruction (BPCO) and were divided into 31 chronic bronchitics (BC) and 20 emphysematous subjects (EP). 10 volunteers who were free of any respiratory pathology were chosen as controls. A visual score for the computed tomography was established for the subjects as a whole. Double reading of the data enabled the reproducibility of the method to be checked in 10 subjects (r = 0.98, p < 0.001). No emphysema was found in the 10 controls, the computed tomography score was appreciably more elevated in the EP subjects than in the BC group at 1.3 and 0.44 respectively (p < 0.001). In the BC, the computed tomographic score was not correlated with the PaO2 (r = 0.54, p < 0.001) and the FEV1 (VEMS) (r = < 0.44, p < 0.05). On the other hand, in the PE group, the score was correlated with the FEV1 (r = 0.52, p < 0.05) and the residual volume (r = 0.06, p < 0.05) and the total lung capacity (r = 0.63, p < 0.05) and the TLCO (r = 0.56, p < 0.05) and the TLCO/VA (r = 0.59, p < 0.05). The adoption of a visual computed tomographic score enabled the authors to find the correlations between pulmonary emphysema and the most specific tests of respiratory function. PMID- 8235022 TI - [The Quebec system of indemnification for occupational asthma. Description, efficacy, and costs]. AB - This study describes the Quebec system of compensation for occupational asthma, assessing the functional and social outcome of claimants and estimating the efficiency and cost. Information was obtained on the clinical, functional and social outcome as well as the estimated costs for 134/211 subjects (participation rate of 64%), who received compensation between 1986 and 1988. At the time of assessment (2 years and more after the diagnosis), 93% of participants still demonstrated significant bronchial hyperresponsiveness and 84% required anti asthma medication. None of the participants remained exposed to the offending agent: 67% were working for the same or another employer, 16% were retired, 8% were retraining for a new job and 8% were still unemployed. Quality of life was mildly affected, more so than for a control group of subjects. The mean interval between the time claims were addressed and the first medicolegal decision was 8.1 months. The mean total cost (including temporary and permanent disability indemnities, medical and technical costs) was $CAN 49,200 (minimum and maximum values of $2,100 and $330,900). We conclude that for subjects with occupational asthma in Quebec: 1) the mean interval for a medicolegal decision to be made is eight months: 2) a minority is still unemployed two to four years after being assessed; 3) the quality of life is more affected than in a control group; 4) the mean cost is close to $CAN 50,000. PMID- 8235023 TI - [Invasive pulmonary aspergillosis in 4 patients with acute decompensation of chronic respiratory insufficiency]. AB - Invasive pulmonary aspergillosis (API) is a necrotising pneumonia generally occurring in profoundly immunodepressed subjects. These observations were based on four patients in the intensive care unit, suffering from chronic respiratory failure (IRC), without profound immunodepression. After a pathophysiological and clinical review, a focus on the diagnostic methods permits one to stress on the reliability, in this type of patient, of the evidence from direct examination of aspergillus filaments in the bronchoalveolar lavage (LBA) or protected bronchial brushings, taking account of the weak value of routine culture of spit or bronchial aspiration in IRC in whom patients are frequently colonised. These four cases permit one to discuss the factors which predispose to the development of API outside the usual immune suppression: IRC itself, by the disorder of mucociliary function, which it leads to; repeated antibiotic therapy which destabilises the saprophytic flora; viral infections which would be responsible for transitory immunodepression. But it is above all steroid therapy which seems to be the major factor favouring the development of API without producing profound immunodepression but probably because it inhibits phagocytosis of aspergillus spores. In these circumstances it is necessary to make an early diagnosis and to use fibre optic bronchoscopy with protected sampling and bronchoalveolar lavage with a complete microbiological. Only early treatment allows one to contemplate a cure. PMID- 8235024 TI - [Prognosis of repetitive excisions of bronchial cancer]. AB - The indications for repeated excision for bronchial cancer may be raised in cases of a synchronous second (or subsequent cancer) or in the case of a local recurrence of the initial cancer which was operated upon. Ninety consecutive patients had a repeat operation for cancer over a 30 year period. There were 86 men and 4 women with a mean age of 59. The initial cancer had most often been treated with lobectomy or bilobectomy and was relatively localised (90% stage I or II). However, 7 patients had a pneumonectomy. The repeat operation was carried out for 30 recurrences and in 44 for secondary subsequent cancers and in 16 cases for a bilateral synchronous cancer. The second operative intervention led to 30 pneumonectomies, 29 lobectomies and 20 atypical excisions. On the otherhand 11 patients had an exploratory thoracotomy and on 10 occasions there was local regional recurrence. These secondary localisations were classified as 44 stage I, 9 stage II and 26 stage III. Eight patients had major loss of lung parenchyma with a pneumonectomy on one side and an atypical excision or a lobectomy on the other. Six patients (6.5%) died in the post-operative period (two from respiratory failure and two due to cardiovascular causes and two sepsis). The actual survival at 5 years was 20% in cases that had an excision. The prognosis was identical in cases having a resection for a recurrence or for secondary subsequent cancer or for bilateral synchronous cancers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235025 TI - [Environmental mesotheliomas in northeast Corsica]. AB - Since 1980, we have collected fourteen cases of mesothelioma induced by environmental exposure to asbestos, going back to childhood in patients from north-east Corsica, in a region which was remote from the asbestos mine of Canari. There were eight men and six women with a mean age of 69.5 +/- 4 years. Six patients presented with bilateral calcified pleural plaques as evidence of environmental exposure. The mineral analysis carried out on five patients (four had thoracoscopies and one an alveolar lavage), showed a moderate deposit of chrysotile (0.3 to 3.4 x 10(6) fibres per gram of dry tissue), and elevated level of tremolite (1.4 to 62 x 10(6) fibres/g). The ambient dosage of asbestos has confirmed the existence of environmental pollution by chrysotile fibres and above all by tremolite. In addition, the same type of fibres have been identified in the parietal pleural of animals subjected to the same risk. In this region, the risk is estimated, on the basis of our results, as 10 cases of mesothelioma per 100,000 inhabitants per year. PMID- 8235026 TI - [Allergic bronchopulmonary aspergillosis and cystic fibrosis. 2 case reports in adults]. AB - The authors report two cases of allergic bronchopulmonary aspergillosis complicating cystic fibrosis in adults: respectively an acute and an exacerbation stage. Association cystic fibrosis-allergic bronchopulmonary aspergillosis is not uncommon and should be considered in all patients with cystic fibrosis especially with atopic disorders. These two cases are a reminder of the diagnosis criteria (clinical, radiologic and biological) and treatment. PMID- 8235027 TI - [Disseminated infection due to an unusual strain of Nocardia farcinica]. AB - Nocardia farcinica, isolated by Nocard in 1888, has recently been redefined as a separate species to asteroides and has been recognised as human pathogen since 1975. Following airborne contamination, lung infection is most usual with frequent dissemination especially to central nervous system. Usually drug resistance makes treatment difficult. The authors describe the first case of Nocardia farcinica disseminated infection in France with lung primarily affection, empyema and encephalitis. PMID- 8235028 TI - [Problems related to thoracoscopic resection of pulmonary nodules]. AB - We present the initial results fo thoracoscopic resection of pulmonary nodules, based on a report of 23 patients. The resection was performed through a sole thoracoscopy in 12 cases, but was associated with a mini-incision in 11 cases. In 4 cases, the procedure had to be converted to thoractomy. We discuss the problems encountered and the indications of the technique. PMID- 8235029 TI - [Pulmonary involvement of Bourneville's tuberous sclerosis. The value of early detection. A familial form]. AB - Pulmonary disease is very rare during the course of tuberous sclerosis of Bourneville (STB). The authors report two cases of STB with pulmonary involvement occurring in the same family, mother and daughter. Both presented with typical cutaneous manifestations of the disease and bilateral renal angiomyolipomas. In the daughter, the early pulmonary diagnosis was made by computed tomographic examination (TDM) which showed the images of the cyst very sharply, although the pulmonary radiograph was normal. Prolonged follow up with pulmonary function tests is important. Lung function tests were very abnormal in the mother with a frank diminution of the TLCO and hypoxia at rest. In the daughter, they revealed the development of obstructive airways disease. Bronchoalveolar lavage was carried out in both the mother and daughter and showed intra-alveolar haemorrhage (with a ground glass appearance on computed tomography in the mother). Pulmonary lymphangiomyomatosis (LPM) and STB with pulmonary involvement are clinical disorders which are anatomically closely related. If the value of hormonal treatment has been shown during the course of LMP, their efficacy in STB is variable. PMID- 8235030 TI - [Mediastinal bronchogenic cyst associated with a multiple malformation syndrome]. AB - The authors report a case of a patient presenting with a bronchogenic mediastinal cyst, which presented as a super-infection of the cyst. The cyst was associated with malformation of the cervical vertebra, cardiovascular abnormalities and congenital deafness, placing this in the group of rare complex polymalformation syndrome with Klippel-Feil Syndrome. The authors also stress the value of computed tomography, and above all of nuclear magnetic resonance in the analysis of mediastinal tumours and in particular, of bronchogenic cysts. PMID- 8235031 TI - [Good health (III). A health structure in southern Italy. A report on its productivity]. PMID- 8235032 TI - [Cardiac abnormalities in systemic lupus erythematosus and their association with antiphospholipid antibodies]. AB - An echocardiographic study was performed in 60 unselected patients affected with Systemic Lupus Erythematosus (SLE) and in 30 age- and sex-matched control subjects, to evaluate the incidence of cardiac abnormalities and the possible correlation with the presence of antiphospholipid antibodies. 33 patients (55%) had major cardiac abnormalities as valvular stenosis and/or regurgitation (35%), pericardial effusion or thickening (26%), left ventricular hypertrophy (21%), regional or global left ventricular dysfunction (10%). Some patients presented more than one cardiac dysfunction: five patients had one major and one minor lesion, 11 presented with two major lesions, and in five of them a pancarditis was found. Minor cardiac abnormalities as mitral valve prolapse, valvular thickening without valvular dysfunction, calcification of the mitral annulus were demonstrated in nine patients (15%). Increased levels of antiphospholipid antibodies were found in 25 out of 60 patients (41.6%). No clear correlation was evident between endocardial or pericardial involvement and such autoantibodies. In fact, valvular heart diseases were present in 36% of the patients with antiphospholipid antibodies and in 34% of the patients without antiphospholipid antibodies; pericardial involvement was evident in 24% and in 28% of patients with and without them, respectively. On the contrary the demonstration of antiphospholipid antibodies in five out of six patients with regional or global left ventricular dysfunction could suggest a pathogenetic role of these autoantibodies in myocardial hypokinesis. Therefore, antiphospholipid antibodies could represent only one of the pathogenetic factors of the cardiac lesions in SLE patients, together with immunologic and iatrogenic factors. The involvement of other systems as renal, vascular and pulmonary certainly play an important role in predisposing to the development of secondary cardiac manifestations. PMID- 8235033 TI - [Liver involvement in systemic lupus erythematosus]. AB - From the revision of the literature it emerges that liver disease in systemic lupus erythematosus (SLE) has been reported in 8 to 23% of the patients and usually is of modest clinical relevance. The incidence of liver abnormalities in 86 patients affected by SLE (9 men and 77 women, mean age 42.1 +/- 13.1 years) was therefore analysed. In our study, liver involvement was defined when at least two abnormal liver function tests in two different occasions were detected. Evidence of liver disease was found in 20 of our patients (23.2%); in 6 cases liver abnormalities were not directly caused by SLE: in 3 subjects the abnormalities found were induced by drugs, in 2 patients by infection and in the last one by fatty liver. In the remaining 14 cases (16.2%), whose hepatic involvement was really due to SLE, a higher frequency of positivity for anti-ds DNA antibodies than in the other patients was observed (92.8% and 51.3% respectively in the two groups, p < 0.05). As not reported previously, these data suggest that liver disease is a prerogative feature of severe SLE. From our casistic in only 1 case the hepatic injury was the onset clinical manifestation of the disease and was clinically relevant as it was diagnosed as chronic active hepatitis. These observations therefore offer the opportunity to discuss the relationship between SLE and autoimmune hepatitis. PMID- 8235034 TI - [Long-term treatment with i.v. immunoglobulin in the therapy of systemic lupus erythematosus]. AB - BACKGROUND: Treatment of several autoimmune diseases with intravenous gammaglobulins (IvIg) has been demonstrated to be safe and effective in determining clinical improvement and decreasing autoantibody titer. We tested the hypothesis that IvIg would be effective in patients with Systemic Lupus Erythematosus (SLE). METHODS: We conducted an open trial involving 12 patients with SLE refractory to conventional treatments with monthly infusion of IvIg at a dosage of 400 mg/kg/day for 5 consecutive days. The duration of the therapy with the same dose every 4 weeks was from 16 to 24 months. RESULTS: A progressive clinical improvement was observed in 11 patients during IvIg therapy, and it persisted during the all period of treatment. The clinical improvement was associated with an increase of haemoglobin, albumin levels, total serum complement and C3 and C4 components, platelets count in 2 thrombocytopenic patients, and a progressive reduction of ESR, plasma immunocomplexes and antinuclear antibodies. A marked improvement in serum urea, creatinine clearance and proteinuria was also observed in the patients with renal involvement. We have not observed any adverse effects with the long-term use of this treatment. Several mechanisms have been postulated to explain the effect of IVIg in the treatment of autoimmune diseases. Many experimental and clinical data support the interesting hypothesis that IVIg may be effective in some autoimmune diseases by restoring a normal function or the physiological immune network through the same regulatory mechanisms leading to a suppression of autoimmune disorders in normal individuals. The presence of a wide spectrum of anti-idiotypic antibodies in IVIg could regulate T and B-cell activities preventing the emergence of B-cell clones with specificity for autoantigens epitopes and down-regulating autoantibodies production. CONCLUSION: The results obtained suggest that IvIg therapy seems to be a promising and beneficial approach in the treatment of SLE. However, double blind studies are necessary to confirm the results obtained particularly to ascertain the optimal dosage, the schedule of infusion and the duration of maintenance therapy with IVIg, and to determine their long-term effectiveness, the reduction in late morbidity and mortality and the effect on the quality of life of patients with SLE. Moreover, because of the high cost of IVIg, their use seems to be especially indicated for patients non responders to conventional treatments and for these with infectious complications. PMID- 8235035 TI - [Acute adrenal failure due to adrenal hemorrhagic necrosis secondary to heparin induced thrombocytopenia]. AB - The authors describe a clinical case with a peculiar sequence of unhealthy events. An operated by osteotomy woman presented a deep venous thrombosis of lever lower extremity with following pulmonary embolism. The patient was treated with heparin. After 5 days, the patient showed a thrombocytopenia, that was not determined by an immune mechanism. The heparin was stopped and the thrombocythemia returned to normal values. But the patient still presented somnolence, asthenia and hypotension. The tests of adrenocortical function showed below normal values. The abdominal CAT showed haemorrhagic necrosis of the suprarenal glands. PMID- 8235036 TI - [New horizons in medicine. Complexity and predictability in internal medicine]. AB - Being a highly sophisticated structure, the human body should be considered on a physical level as a whole of inter-correlated non-linear dynamic systems, which are by definition, complex systems because they are always conditioned in their operativity by numerous variables. We must draw attention to the opportunity that within the field of medical science the concept of complexity referring to diagnostic, therapeutic and prognostical problems, is distinct between structural and functional. The functional complexity of a system, a phenomena, an event, MY be recognized in its dynamics, only if analysed using modern methods recently brought to light, on the relationship which exist between classic determinism and deterministic chaos. Furthermore, also on practical level, the recent discoveries on the general principles, which are at the base of the functional complexity of non-linear dynamic systems, demonstrate exactly how fragile is the probabilistic predictability which guides a doctor in clinical reasoning in all its various stages; in the gathering of anamnestic and objective data; in the programming the researches aimed at confirming the validness of the diagnosis; in the therapy aimed at reaching the best possible results; in the prognosis which summarize the destiny of the binomial patient illness. The scientific enunciate of universal nature, which demonstrate how, from a tiny uncertainty in the initial functional data of a system, of a phenomena, of an event can unleash a condition of absolute unpredictability, find daily confirmation in both clinical and experimental medicine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235037 TI - [Alcoholism. Aspects of its pharmacology, clinical picture and therapy]. AB - In every age and culture, people have had to distinguish between positive and negative consequences of the use of psychoactive substances. Since the ancient times it was known that alcohol, easily available in wide quantities, could facilitate social contacts. The abuse and the addiction represent, on the other hand, a complicated social phenomenon and are the expression of physiological adaptive mechanisms brought to the extreme point. The high level of diffusion and the complexity of these problems made it necessary to create diagnostic classifications to classify illnesses on the basis of definite and reliable principles, with the main goal of obtaining therapeutic progress. The consequences of alcohol abuse are both physical and psychic, thus therapeutic approaches must take into account these two points. Some pharmacological agents may lead to cross-tolerance with alcohol and, besides, they are not necessary in case of a slight abstinence where brief psychotherapies are sufficient. Recent advances in the knowledge of the mechanisms which underlie alcohol addition suggest approaches particularly aimed to correct imbalances of neurotransmitters that can determine addiction. Among these, GHB acid seems to be particularly important. At the end, on the basis of the latest progress in psycho immunopharmacology, the relationships between alcohol and disease are considered. PMID- 8235038 TI - [The visualization of the secondary lobule by high-resolution CT: the semeiological implications in the compartmental definition of changes]. PMID- 8235039 TI - T-cell recognition of superantigens. PMID- 8235040 TI - Analyses of the staphylococcal toxin binding sites on MHC class II molecules. PMID- 8235041 TI - Endogenous superantigens: tools for dissecting T-cell repertoire selection. PMID- 8235042 TI - Roles of superantigens in microbial infections? AB - Superantigens have been defined in a variety of infectious particles such as bacteria and viruses. These superantigens have the capacity to stimulate a large percentage of the host T cells by interacting specifically with the T-cell receptor V beta chain which is shared by about 1-20% of mature T cells. The recent discovery that mammary tumour viruses express such superantigens enabled the analysis of the retroviral life cycle and led to questions about the role of superantigen in amplification of the infection. PMID- 8235043 TI - The fringes of superantigen reactivity: what do they tell us? PMID- 8235044 TI - Superantigens. PMID- 8235045 TI - Rabies virus superantigen. PMID- 8235046 TI - Parental influences on childhood fitness and activity patterns. AB - The purpose of this study was to determine the effect of parental attitudes and self-reported exercise habits on the fitness and activity levels of their children. One parent from each of 1,253 families was asked about exercise habits and completed the Exercise Benefits and Barriers Scale (EBBS) to determine their personal attitudes toward exercise. The children completed a self-reported activity questionnaire (SRA) and had their aerobic power predicted using a cycle ergometer test. Multiple regression indicated that the parents' EBBS scores were weakly associated with the child's maximal oxygen uptake (VO2max); the mothers' association was significant, but the fathers was not. The children's SRA scores were not correlated with parents' attitudes or exercise habits. These results suggest either that factors other than parental attitudes and exercise habits are more influential in determining the fitness and activity levels of children or that the instruments lack precision. Thus, to improve the cardiovascular fitness of children, we need to examine factors in the child's environment that may be more influential than parental attitudes and examples. PMID- 8235047 TI - The multidimensional structure of physical fitness: invariance over gender and age. AB - This investigation extends the factor analytic approach pioneered by Fleishman (1964) by incorporating subsequent developments in the application of confirmatory factor analysis and the physical fitness literature. Specifically, it tests the ability of an a priori factor structure of physical fitness to fit (i.e., account for) data based on 25 indicators of fitness (field exercises, technical measures, and laboratory measures) for 2,817 boys and girls aged 9, 12, and 15. An eight-factor (Cardiovascular Endurance, Explosive/Dynamic Strength, Static Strength, Flexibility/Joint Mobility, Blood Pressure, Lung Function, Body Girth, and Skinfold) model derived from previous research fit the data well for each of the six age/gender groups, considered separately. Based on tests of factorial invariance, factor loadings and factor correlations were reasonably invariant across the six groups. This important finding indicates that all 25 indicators are equally valid for boys and girls aged 9, 12, and 15 and that the multidimensional structure of physical fitness generalizes over gender and age. PMID- 8235048 TI - Rapid movement kinematic and electromyographic control characteristics in males and females. AB - Maximally fast, self-terminated elbow flexion movements were performed by 10 male and 10 female college-aged subjects to assess potential gender-related differences in kinematics and the triphasic electromyographic (EMG) pattern. The subjects were instructed to move their forearms as fast as possible through 90 degrees of elbow flexion range of motion and stop as sharply as possible at the terminal point. An electromagnet, set to 0, 40, and 70% of each subject's maximal isometric torque, provided resistance to movement initiation and resulted in quick release movements. Surface EMG was collected from the biceps b. and triceps b. muscles. Results indicated that the males had faster movements and accelerations under all conditions. EMG records indicated that the males had faster rates of EMG rise, particularly in the triceps b., and more tightly coupled reciprocal activation. The quick release afforded faster accelerations for both groups, yet only the males moved faster throughout the full range of motion. Following the quick release, the males differed from the females by increasing the triceps b. EMG amplitude. Hence, the males were able to shorten movement time in quick release movements by increasing triceps b. activation and, thus, braking ability. These results suggest that the females were more neurally constrained than the males with respect to rapid EMG activation of the triceps b., resulting in limits in the braking process. PMID- 8235049 TI - Rhythmicity, ritual, and motor performance: a study of free throw shooting in basketball. AB - The purpose of this study was to determine the importance of timing during an auto-communicative ritual to successful performance. Eight members of a university varsity basketball team served as subjects for this study. Each subject performed 15 free throws in each of four different conditions. Condition 1 required subjects to use their standard free throw ritual prior to shooting. Condition 2 required subjects to maintain the relative timing of ritual behaviors but reduce the absolute time of the ritual by one-half. Condition 3 required that relative timing be maintained but that the absolute time of the ritual be doubled. Condition 4 required the same behaviors and the same absolute time as the standard ritual but that the relative time of the ritual be altered. Data were collected with a 16-mm high-speed camera. Types of behaviors, timing aspects of the ritual, successful attempts, and shot mechanics were dependent measures. Analysis of variance (ANOVA) of dependent measures and Fisher's scores from correlation coefficients of dependent measures indicate that relative timing of behaviors (rhythmicity) is more important to success than the absolute time of rituals. Behaviors most important to free throw success are those that may be totally controlled by the subject and yet remain stable with changing conditions. PMID- 8235050 TI - A Pathfinder analysis of pedagogical knowledge structures: a follow-up investigation. AB - This study is an extension of an earlier investigation of undergraduate students' acquisition of key pedagogical concepts in a physical education teaching methodology course. In that study, Pathfinder, a method for eliciting associative memory networks, was used to describe and compare the pedagogical knowledge structures of students to that of the course instructor. After the course, students' pedagogical knowledge structures corresponded more closely with that of the instructor, and students who corresponded the closest performed better in the course. The results raised an interesting issue regarding the acquisition of knowledge in undergraduate students. Did students acquire a generalizable body of pedagogical knowledge applicable beyond the context of the teaching methodology course or a highly contextualized reflection of their course instructor's knowledge base? In the present study the external validity of the pedagogical knowledge base was examined by using Pathfinder to compare the knowledge structures of students from the initial investigation with knowledge structures of five experienced teacher educators from five different teacher education programs. The findings indicated that students' knowledge structures became significantly more correspondent with that of the experienced teachers' structures from the beginning to the end of the course. Also, students' correspondence with teacher educators' structures following instruction was found to be significantly correlated with academic and teaching performance. The findings point to the external validity of the domain of knowledge under study and the robustness of Pathfinder for capturing pedagogical knowledge. PMID- 8235051 TI - Physiological responses to maximal treadmill and handweighted exercise. AB - The purpose of this investigation was to compare the peak physiological responses among four protocols that employed different amounts of handweighted exercise in 16 males (aged 26.3 +/- 4.1 years). The four protocols were (a) uphill treadmill running (UR; 3.36 m.s-1, 2.5% grade increase-3 min-1); (b) uphill treadmill walking while pumping 1.36-kg handweights (HW) (UWHW; 1.79 m.s-1, 5.0% grade increase x 3 min-1; (c) treadmill walking while pumping .91-kg HW (WHW; 1.79 m.s 1, 0% grade, .91-kg HW increase x 3 min-1); and (d) standing in place and pumping HW (SHW; arm work as described in WHW). It was hypothesized that the peak responses would be inversely proportional to the estimated muscle mass activated (i.e., UR = UWHW > WHW > SHW). Dependent variables included peak oxygen uptake (VO2 peak), peak heart rate (HRpeak), peak ventilation (Ve peak), and peak respiratory exchange ratio (RERpeak). No differences were noted between UR and UWHW with respect to any of the dependent variables. All variables (except RERpeak) were greater (p < .01) in UR and UWHW than either WHW or SHW. RERpeak was greater (p < .01) in UR and UWHW than in WHW. VO2 peak and HRpeak were greater (p < .01) in WHW when compared to SHW. Mean VO2 peak was 97.5, 69.7, and 60% of UR for UWHW, WHW, and SHW, respectively. Therefore, walking and pumping handweights provides a maximal stimulus to the oxygen transport system. PMID- 8235052 TI - Exercise and VO2 max in children: a meta-analysis. AB - Despite widespread belief that children are aerobically trainable, studies examining the ability of a child to improve maximal oxygen uptake (VO2max) have yielded inconsistent findings. The present investigation, using meta-analysis, examined the effects of physical activity, gender, experimental design, and sufficiency of exercise on the VO2 max of child subjects. Sixty-nine studies examining the effects of training on children were originally located; 28 met criteria for inclusion. From these studies, 70 effect sizes (ESs) were calculated. Some studies employed cross-sectional (XS) designs involving comparisons of intact groups of subjects; the others used a pretest-posttest (PP) design, which followed subjects throughout a specified training program. Average ES indicated a considerable difference between trained and untrained subjects though several possible sources of confounding (e.g., subject self-selection) in XS studies were identified. Effect sizes of .94 (+/- 1.00) and .35 (+/- 0.82) were achieved for XS and PP designs, respectively. Further analyses were conducted with the PP design studies. In these studies, subjects improved approximately 2 ml.kg 1 x min-1. In the PP studies, effect sizes were not significantly affected by (a) gender, (b) "sufficient" and "insufficient" training protocol, or (c) test mode. Results indicated that reported changes in VO2max in children are small to moderate and are a function of the experimental design used. PMID- 8235053 TI - The role of cognitive appraisal and friendship provisions in adolescents' affect and motivation toward activity in physical education. AB - This study investigated the mediational role of affect in the influence of perceived success, causal attributions, and friendship provisions on choice and future expectancies for success and enjoyment, using a multi-sample structural equation modeling approach. Subjects were 422 seventh- and eighth-grade students participating in physical education classes. Higher levels of perceived success, personally controllable attributions, companionship, and esteem support were found to positively influence levels of affect experienced, which in turn enhanced future expectancies for success and enjoyment in physical education and choices to participate in physical activity outside of school. Furthermore, participation experience outside of school influenced adolescents' perceptions within the physical education setting and their consequent affect and motivation. PMID- 8235054 TI - Effects of same-sex versus coeducational physical education on the self perceptions of middle and high school students. AB - The purpose of this field experiment was to investigate the effects of attending either a coeducational or a same-sex physical education class on several self perception variables. Middle and high school youth who had previously been in coeducational classes were assigned to either a same-sex or a new coeducational physical education class for a 10-lesson unit of basketball. Analyses were conducted at both the group and the individual levels. Self-perception variables examined included perceived self-confidence of learning basketball, perceived usefulness of basketball, and perceived gender-appropriateness of basketball. Results of hierarchical linear model group level analyses indicated that the variability in groups for self-confidence could be explained by grade, class type, and the interaction between gender and class type. At the individual level, multivariate results showed that, after the unit, males in coeducational classes were significantly more confident in their ability to learn basketball than males in same-sex classes. Also, males in same-sex classes decreased in confidence from pretreatment to posttreatment. Perceived usefulness of basketball emerged as the strongest predictor of self-confidence for learning basketball for both genders. In general, middle school students preferred same-sex classes, whereas high school students preferred coeducational classes. PMID- 8235055 TI - Path analysis of a self-esteem model across a competitive swim season. AB - An adaptation of the previously developed Exercise and Self-Esteem Model (Sonstroem & Morgan, 1989) was tested longitudinally with 93 male interscholastic swimmers from nine high schools who were evaluated at pre-, mid-, and postseason. Swimmers completed three self-perception scales that ranged from evaluations of specific swim skills (SKILL) through broader perceived physical competence (PC) to global self-esteem (SE). Performance scores (PERF) were calculated across events by standardizing swim times to a mean of 0 and a standard deviation of 1. Structural modeling analysis across the three time waves indicated an excellent data fit (chi 2 = 25.46, p > .15). The model was able to explain 84, 83, and 80% of PC, SKILL, and PERF, respectively, at the third testing period. Swim improvement tended to be small (median change = 2.9%). Relationships among variables at each testing period and among the same variables at different testing periods were large and as hypothesized. Several significant relationships occurred between different variables across different testing periods. These were not extinguished when social desirability was added to the model. Social desirability failed to significantly influence subsequent measures of any model variable. PMID- 8235056 TI - Relationships between changes in health and fitness and the perception of exercise. AB - This article examines the answers respondents gave to two questions about their perception of recent changes in their health and fitness in relation to the answers respondents gave to questions asking if they thought (a) more exercise would improve their health, (b) they got as much exercise as needed, and (c) they were as active as others of a similar age. A theoretical model is discussed that emphasizes the importance of a change in the perception of health and fitness as a cue to reevaluating exercise and leisure time-related beliefs, attitudes, and behaviors. Data from 9,125 female and 7,157 male respondents collected by telephone in the urban areas of Glasgow and Edinburgh were analyzed. The analysis indicated a strong relationship between individuals reporting a negative change in health or fitness and those individuals being of the opinion that more exercise would improve their health, that they got less exercise than they needed, and were less active than others of a similar age. PMID- 8235057 TI - A preliminary investigation of the effect of weight training on salivary immunoglobulin A. PMID- 8235058 TI - How "significant" is your reliability? PMID- 8235059 TI - Laboratory assessment of choice between exercise or sedentary behaviors. PMID- 8235060 TI - A proposed test for determining physical working capacity at the heart rate threshold. PMID- 8235061 TI - Effect of NS-398, a new nonsteroidal anti-inflammatory agent, on gastric ulceration and acid secretion in rats. AB - The ulcerogenic activity of NS-398 was compared with that of indomethacin and the effects of NS-398 on stress-induced ulceration, gastric acid secretion and gastric mucosal prostaglandin (PG) contents were investigated in rats. NS-398 in a single dose of up to 1,000 mg/kg, p.o. did not significantly cause gastric ulceration while other nonsteroidal anti-inflammatory drugs such as, loxoprofen, indomethacin, diclofenac and ibuprofen, produced distinct gastric lesions. In cases of stress-induced ulcerations, NS-398 at 30 mg/kg, p.o. had no significant influence while indomethacin markedly potentiated the ulceration in a dose dependent manner. In basal gastric secretion studies, both NS-398 and indomethacin decreased secretion volume and acidity. However, in the 2-deoxy-D glucose-stimulated gastric acid secretion study, NS-398 had no significant influence on gastric secretions while indomethacin significantly potentiated the secretion. Both NS-398 and indomethacin to much the same extent significantly decreased prostaglandin E2 (PGE2) contents in inflammatory tissue. However, with respect to gastric mucosal PGE2 contents, NS-398 did not decrease PGE2 contents while indomethacin significantly decreased the contents. It would thus appear that the absence of ulcerogenic properties of NS-398 is due to a relative lack of activity in inhibiting gastric PG synthesis. PMID- 8235062 TI - Plasma angiotensin I concentration and serum placental leucine aminopeptidase (P LAP) activities in puerperal hypertension. AB - To elucidate the involvement of angiotensin in puerperal hypertension with and without preeclampsia in the course of prior pregnancy, we measured plasma angiotensin I levels and serum placental leucine aminopeptidase (P-LAP) activities in several puerperal hypertensive patients. Placental leucine aminopeptidase (P-LAP) activities of puerperal hypertensive patients with severe preeclampsia during prior pregnancy were equal to or somewhat lower levels than those in normal puerperal women after a normal pregnancy. However, their angiotensin I levels were higher than those in normal puerperal women after a normal pregnancy. In puerperal hypertensive patients with and without preeclampsia during prior pregnancy, P-LAP activities tended to show much lower levels that those in normal puerperal women, while angiotensin I levels tended to show much higher levels than those in normal puerperal women, respectively. Since serum placental leucine aminopeptidase acts as an angiotensinase via degradation of angiotensin III (A-III), our present data suggest the involvement of angiotensin in puerperal hypertension. PMID- 8235063 TI - Antitussive effect of captopril, an angiotensin converting enzyme inhibitor, in the rat. AB - The effects of acute treatment with captopril, an angiotensin converting enzyme inhibitor, on the capsaicin-induced cough reflex were examined in rats. Intraperitoneal injection of captopril in doses of 3 and 10 mg/kg decreased the number of coughs dose-dependently. Although the peak effect was similar to that of morphine (0.5 mg/kg), the duration of captopril's effect (10 mg/kg) was longer than that of morphine. Furthermore, while pretreatment with naloxone significantly decreased the duration of the antitussive effect of captopril, it had no significant effect on the early-phase (within 60 min) effect of captopril. These results suggest that the mechanism which underlies the antitussive effect of captopril involves mediation by both nonopioid and opioid systems. PMID- 8235064 TI - Overexpression of HLA class I antigen reduces insulin secretion in pancreatic beta cells (RINM5F): evidence for a non-immune mechanism. AB - Our recent observation indicated that overexpression of HLA-class I antigen on pancreatic beta cells is one of the features of insulin-dependent diabetes mellitus (IDDM). To analyze the effect of class I antigen overexpression, we have introduced human HLA class I (HLA-Cw3) gene into rat pancreatic beta cell lines, RINm5F. Several stable transformants with various levels of surface expression of class I antigens have been cloned. The insulin secretion of these transfectants was analyzed to evaluate the functions of beta cells. Highly negative correlation between the level of insulin secretion and that of class I expression (correlation coefficiency: r = 0.89) has been observed. These data suggest that the overexpression of HLA class I antigen itself may impair pancreatic beta cells through a nonimmune mechanism. PMID- 8235065 TI - Biochemical characterization of flestolol esterase. AB - The blood esterase that mediates the metabolism of flestolol, an ultra short acting beta blocker, was characterized. Esterase activity occurred in plasma of human, dog, rat, and guinea pig and not in erythrocytes of the same species. The esterase activity was greatest in humans and guinea pigs followed by dogs and rats. Purified human serum cholinesterase was very active against flestolol while human serum albumin was slightly active. Human and bovine erythrocyte membrane acetylcholinesterases, electric eel acetylcholinesterase, human hemoglobin, dog, rat, chicken, and bovine serum albumin were all inactive. Esterase activity with flestolol was inhibited in human, dog, and rat blood by echothiophate, eserine, and sodium fluoride. Guinea pig blood esterase activity was inhibited by echothiophate and sodium fluoride, but not by eserine. Metabolic interaction studies indicated that succinylcholine, procaine, and chloroprocaine interfere with the metabolism of flestolol in human blood. Succinylcholine prolonged the in vitro half-life of flestolol in dog blood, but acetylcholine, procaine, and chloroprocaine had no effect. Flestolol did not affect the metabolism of procaine or chloroprocaine in human and dog blood. The metabolism rate of flestolol decreased in individuals with atypical, fluoride-resistant and silent forms of serum cholinesterase. PMID- 8235066 TI - Increased excretion of proline-containing peptides in dipeptidyl peptidase IV deficient rats. AB - We have reported that a substrain of Fischer 344 rat exhibits a deficiency of dipeptidyl peptidase IV (Watanabe et al. Experientia 43: 400-401, 1987). In this study, this rat model was found to excrete large amounts of proline-containing peptides in the urine. The peptide fraction also contained other amino acids, such as hydroxyproline and glycine. The results indicate that the enzyme might be involved in the metabolism of collagens and related peptides. PMID- 8235067 TI - Effect of diacylglycerol kinase inhibitor, R59022 on cytosolic free calcium level and force development in guinea pig taenia coli. AB - We previously described that carbachol (CCh) increased intracellular phosphatidic acid (PA) in guinea pig taenia coli and the increase was inhibited by diacylglycerol (DG) kinase inhibitor, R59022. The present study was conducted to clarify the role of DG kinase in CCh-induced increase in cytosolic free calcium and force development. Application of CCh to taenia coli induced transient increase in calcium level and contraction followed by sustained and lower calcium level and smaller contraction, and R59022 inhibited the latter responses specifically. The sustained responses were also evoked by 50 mM KCl, but these responses were slightly inhibited by R59022. Inhibition of calcium influx reduced responses to CCh but not KCl. Therefore, we consider that the sustained contraction induced by CCh is attributed to activation of DG kinase which generates PA and accelerates calcium influx. PMID- 8235068 TI - Muscle relaxant action of dantrolene sodium in rats. AB - The relationship between the muscle relaxant action and intramuscular concentration of unchanged dantrolene sodium [1-[5-(p-natrophenyl) furfurylidene amino] hydantoin sodium hemiheptahydrate, dantrolene] by intravenous administration of dantrolene was examined in rats. The mechanism of the muscle relaxant action of dantrolene was also examined. Dantrolene, unlike tolperisone, a centrally acting muscle relaxant, dose-dependently inhibited twitch contraction evoked by electrical stimulation of both the muscle and the phrenic nerve in the rat isolated phrenic nerve-diaphragm preparation to a similar extent (IC50: 9.5 and 6.5 x 10(-6) M, respectively). Inhibition of both twitch contractions by dantrolene (50 microM) was antagonized by 4-aminopyridine (42.5 microM) and caffeine (4 mM). The twitch contraction of the gastrocnemius muscle elicited by sciatic nerve stimulation was inhibited by intravenous administration of dantrolene in a dose-dependent manner. The effective intramuscular concentration of dantrolene producing 50% depression (ID50) was 3.4 micrograms/g tissue, equal to about 1 x 10(-5) M. We thus found a close relationship between IC50 in vitro and ID50 in situ. Furthermore, this study raised the possibility that dantrolene produces its muscle relaxant action by affecting peripheral muscular Ca2+ mobilization. PMID- 8235069 TI - Metabolism of allylnitrile to cyanide: in vitro studies. AB - In liver fractions from male Sprague-Dawley rats, the metabolism of allylnitrile (ALN) to cyanide (CN-) was localized in the microsomal fraction and required NADPH and oxygen for maximal activity. The biotransformation of ALN to CN- was characterized with respect to time, microsomal protein concentration, pH and temperature. Metabolism of ALN was increased in microsomes obtained from phenobarbital-treated rats (160% of control) and decreased with cobaltous chloride and beta-diethyl aminoethyl-2,2-diphenyl pentanoate (SKF 525-A) treatments (48% of control). Addition of SKF 525-A to the incubation mixtures inhibited ALN metabolism to CN-. Addition of the epoxide hydrolase inhibitor, 1,1,1-trichloropropane 2,3-oxide, decreased the formation of CN- from ALN. Addition of glutathione, cysteine, D-penicillamine, and 2-mercaptoethanol enhanced the release of CN- from ALN. These findings indicate that ALN is metabolized to CN- via a cytochrome P-450-dependent mixed-function oxidase system. PMID- 8235070 TI - Low response of DNA synthesis upon growth stimulation in LEC mutant rat. AB - Long-Evans Cinnamon (LEC) mutant rat, which spontaneously develops a chronically necrotizing hepatic injury at 4 months of age, exhibits an excess hepatic copper accumulation. DNA synthesis upon growth stimulation in LEC rat primary-cultured hepatocytes markedly decreased when compared to that of normal rat hepatocyte culture. Low response of DNA synthesis seems to be associated with nuclear disorganization induced by copper toxicity. However, LEC rat electrophoretic profile and content of each histone component responsible for DNA stability were indistinguishable from those of normal rat. These results suggest that copper generated damage of substrates other than histones causes nuclear damage in LEC rat. PMID- 8235071 TI - Effects of ferric citrate injection on the composition of bile acids in rats. AB - Ferric citrate was examined for its possible induction of acute and chronic changes in bile acids in rats. Three hours after a single intravenous Fe(III) injection both the biliary bile acid level and the percentage of cholic acid in bile were significantly higher in treated rats than in control rats. However, no significant difference was observed after treatment with the same daily dose for one week. The superoxide dismutase (SOD) activity in liver tissue increased in the chronic group. These results suggest that oxygen free radicals, mainly the hydroxyl radical, promote hydroxylation during bile acid biosynthesis by enhanced monooxygenase activity. PMID- 8235072 TI - Fibroblast migration and proliferation during in vitro wound healing. A quantitative comparison between various growth factors and a low molecular weight blood dialysate used in the clinic to normalize impaired wound healing. AB - During the formation of granulation tissue in a dermal wound, platelets, monocytes and other cellular blood constituents release various peptide growth factors to stimulate fibroblasts to migrate into the wound site and proliferate, in order to reconstitute the various connective tissue components. The effect on fibroblast migration and proliferation of these growth factors, and of Solcoseryl (HD), a deproteinized fraction of calf blood used to normalize wound granulation and scar tissue formation, was quantified in vitro. The presence of basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), transforming growth factor-beta (TGF-beta) and hemodialysate (HD) increased the number of cells in the denuded area, i.e., in the "wound space" of an artificially ruptured monolayer of LM-fibroblasts (mouse lung fibroblasts). When cell proliferation was blocked with Mitomycin C, in the first 24 h all factors, i.e., bFGF, PDGF, TGF-beta and HD, promoted cell migration, whereas after 48 h it became obvious that each factor stimulated both migration and proliferation, each in a characteristic way. The effects were significant and more distinct after 48 h, following the order: PDGF (46%) approximately bFGF (87%) > HD (45%) approximately TGF-beta (40%) > control (62%). The relative contributions of migration after inhibiting proliferation are given in brackets. The modulatory activity of HD was localized in its hydrophilic fraction. It was destroyed by acid hydrolysis. Furthermore, this activity could be blocked by protamine sulfate, an inhibitor blocking peptide growth factor receptor binding. PMID- 8235073 TI - Ketamine-diazepam N2O combination anesthesia--a new "circulatory-system-sparing" type of anesthetic in experimental surgery. AB - For studying the hemodynamics of the greater and lesser circulatory system in animals, the impact of general anesthesia upon cardiovascular functions should be as small, as possible. The marked circulation-depressing effect of halothane, which has so far been given preference for experiments on large animals, was confirmed by the present study of the miniature pig. There was a significant decrease in MAP, cardiac index and dp/dtmax/IP, and an increase in PAPp. Useful insights into hemodynamics, derived from experiments modeling the heart, lungs or vascular system cannot be gained for a cardiovascular system, the functions of which are greatly altered by such an anesthesia. As a new form of anesthesia with minimal cardiovascular influence, the ketamine-diazepam N2O combination, which has already been successfully applied in operations on the open human heart, is introduced into experimental surgery. PMID- 8235074 TI - The adaptive response of the rat small intestine after resection and segmental transplantation during the early postoperative phase. AB - Organ harvesting from a living donor or spatial constraints in the recipient's abdominal cavity are the main factors to be considered in the segmental transplantation of the small intestine. It was the aim of the following study to gain insight into the functional characteristics of different portions of the small intestine either after partial resection or syngeneic and allogeneic transplantation during the early postoperative period. 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 BW s.c.) was administered for graft acceptance. Controls were unoperated, 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 the three portions investigated (P < 0.01) but did not affect maltose absorption. Excretion of fecal fat was elevated 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. Postmortem examination revealed no signs of acute rejection. When transplantation of short intestinal segments is considered, it is of vital importance to take into account the functional differences and the influence of immunosuppressive drug therapy in the regulatory bowel function. PMID- 8235075 TI - Increased endothelin plasma concentrations in patients with coronary artery disease or hyperlipoproteinemia without coronary events. AB - Endothelins are made by endothelial cells, macrophages, and vascular smooth muscle cells, among others, and are the most potent endogenous vasoconstrictors yet discovered, with additional growth-promoting properties. A locally increased endothelin production in coronary artery disease or other atherosclerotic diseases may increase circulating endothelin plasma concentrations before symptoms of disease are manifest. We determined endothelin plasma concentrations (1) in 43 patients suffering from coronary artery disease (CAD); (2) in 43 patients with hyperlipoproteinemia without coronary artery disease (HLP); (3) in 29 healthy control subjects (C), by means of a novel extraction procedure and radioimmunoassay followed by chromatographic separation. Plasma concentrations in HLP and C overlapped, but were still significantly different (29 +/- 10 vs 21 +/- 8 fmol/ml, ANOVA and Duncan's test). Significantly increased plasma concentrations were also found in patients with CAD, with the highest levels in a subgroup of 8 patients presenting with unstable angina (43 +/- 12 vs 53 +/- 15 fmol/ml). There were no statistically significant differences between CAD groups with (n = 28) or without hyperlipoproteinemia (n = 15) (42 +/- 14 vs 41 +/- 16 fmol/ml; n.s.). Likewise there was no relationship between endothelin plasma concentration in any of the patients studied and lipid fractions in serum. Increased endothelin plasma concentrations in HLP patients without evidence of coronary artery disease are thus not related to the hyperlipidemic state per se, but may rather indicate presence of an increased vasoconstrictor tonus, pre clinical or silent atherosclerotic disease. PMID- 8235076 TI - Portal vein infusion of cancer chemotherapeutic agent emulsified with Lipiodol in regenerating liver after partial hepatectomy in rats. AB - Portal vein infusion of aclarubicin (ACR) emulsified with Lipiodol (LP) in regenerating liver after 70% partial hepatectomy in rats was evaluated for its safety and usefulness. DNA synthesis in regenerating liver was transiently suppressed by LP, ACR or LP+ACR, but no obvious inhibition was seen in aminopyrine N-demethylase activity and liver weights. LP significantly enhanced hepatic tissue levels of ACR and its active metabolites by long-term retention in the sinusoidal space. This study demonstrates that in rats LP has a powerful effect on the long-term retention of anticancer agents in the sinusoidal space after infusion into the portal vein, without aggravating hepatic damage by anticancer agents. PMID- 8235077 TI - Metabolic and haemodynamic effects of dopamine plus domperidone in volunteers. AB - There are no studies of the relationship between infusion rate of dopamine and the arterial and venous dopamine plasma concentration and the resulting haemodynamic and metabolic effects. Dopamine was administered to seven volunteers using five infusion rates (1, 3, 6, 9, 13 micrograms/kg per minute) in an escalating sequence lasting for 30 min for each step. Since dopamine can cause nausea and vomiting, this relationship was investigated after administration of domperidone for infusion rates above 3 micrograms/kg per minute. Haemodynamic effects were assessed using 2-dimensional echocardiography. During the highest infusion rate the arterial plasma dopamine concentration reached 1,379 +/- 181 nmol/l. There was a linear correlation between the dopamine infusion rate and both the arterial and the venous plasma concentration. There was no significant change in heart rate or diastolic blood pressure. Systolic blood pressure, ejection fraction and cardiac index increased in a dose-dependent manner. Systemic vascular resistance decreased during the two low doses of dopamine and was not different from baseline values during the three high infusion rates. The plasma concentrations of glucose and non-esterified fatty acids increased from 5.3 +/- 0.4 to 0.68 +/- 0.9 nmol/l, and from 360 +/- 119 to 971 +/- 307 mumol/l, respectively, during the 13 micrograms/kg per minute infusion rate. As the plasma noradrenaline concentration increased up to 7.84 +/- 2.46 nmol/l in correlation to the dopamine plasma concentration, an indirect sympathomimetic effect may contribute to the actions of dopamine plasma concentration. PMID- 8235078 TI - Lung compliance, lung volume and transfer factor for carbon monoxide in anaesthetised sheep: normal values and reproducibility of measurements. AB - Measurements of quasistatic compliance (Cqst), effective alveolar volume (VA,eff) and single-breath transfer factor for carbon monoxide (TL,CO, 'sb') were completed in 16 normal, anaesthetised, adult Texel ewes. Regression equations were computed for these variables as a function of bodyweight and the optimal equations selected. The 95 per cent prediction intervals for the equations were calculated such that normal lung function in similar sheep could be accurately predicted. The long term reproducibility of these measurements was assessed in nine sheep, measured at monthly intervals over a period of five months. Although measurements made in individual sheep were often highly variable, the variation between repeated measurements on the separate days for the group was insignificant. PMID- 8235079 TI - An experimental study of cross-circulation for cardiopulmonary bypass in dogs. AB - Cardiopulmonary bypass using cross-circulation was evaluated in dogs. In eight experiments a greyhound (the donor) was used to perfuse a small mongrel dog (the recipient). Recipients averaged 40 per cent of the donor's weight. Total cardiopulmonary bypass was successfully established in all eight experiments and maintained for 60 to 90 minutes. Haemodynamic values (mean arterial pressure, central venous pressure, cardiac output) and arterial blood gas tensions in the recipient and donor dogs remained at near-baseline levels for the duration of cardiopulmonary bypass. Seven of the eight recipient dogs were successfully weaned from cardiopulmonary bypass at the end of the procedure. PMID- 8235080 TI - Furosemide magnifies the exercise-induced elevation of plasma vasopressin concentration in horses. AB - The purpose of this study was to test the hypothesis that furosemide administration before exercise would cause greater increases in plasma arginine vasopressin (AVP) concentration in exercising horses than exercise alone. Six adult, clinically normal, unfit mares underwent three randomly ordered 60 minute standard exercise tests on an equine treadmill to examine the effect of furosemide administration on plasma AVP concentration. In one trial, furosemide (1 mg kg-1) was infused four hours before exercise (FUR-4) and a placebo (10 ml saline) was infused two minutes before exercise; in another trial the placebo was infused four hours before exercise and drug was infused two minutes before exercise (FUR-2); in a third trial a placebo was infused four hours and two minutes before exercise (CON). During the treadmill test each mare ran up a fixed 4 degrees incline for one hour at a speed previously determined to produce a heart rate of 65 per cent of each horse's maximum heart rate. Venous blood samples were collected at rest in the stall, immediately before exercise while standing on the treadmill, and at 15 minute intervals during the treadmill test. Plasma AVP concentration was measured by radioimmunoassay. In the CON trial, plasma AVP concentration increased 561 per cent (P < 0.05) from 6.3 +/- 1.0 pg ml 1 (mean +/- SE) at rest to 38.8 +/- 12.8 pg ml-1 at the end of the 60 minute run. During the FUR-2 trial, AVP increased 1185 per cent (P < 0.05) from 5.9 +/- 1.7 pg ml-1 to 75.8 +/- 17.7 pg ml-1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235081 TI - Effects of papain on the agglutination of canine red cells with serum autoantibodies. AB - The papain test, which detects incomplete anti-red blood cell (RBC) autoantibodies in serum, was positive in 12 out of 16 anaemic dogs. Positive results were significantly correlated (chi 2 = 11.1, P < 0.001) with increased levels of RBC-bound immunoglobulin, but in three of these cases it was considered that a diagnosis of autoimmune haemolytic anaemia could not be justified. Furthermore, enzyme-linked antiglobulin test measurements of RBC-reactive serum IgG were increased in only three of the dogs with a positive papain test. Papainised canine RBC did not consistently take up more serum IgG than untreated cells in an indirect enzyme-linked antiglobulin test. However, the zeta potential of the RBC was reduced after enzyme treatment, and electrophoretic analysis revealed that glycophorins, which bear a strong negative charge, were cleaved from the cell membrane. It is concluded that a positive papain test alone is not reliable in the diagnosis of canine autoimmune haemolytic anaemia and that the enzyme increases the agglutinability of RBC by reducing the mutually repulsive electrostatic forces between the cells, rather than by increasing the amount of autoantibody bound. PMID- 8235082 TI - Evaluation of free or liposome-encapsulated ribavirin for antiviral therapy of experimentally induced feline infectious peritonitis. AB - Ribavirin, either free in aqueous solution or incorporated into liposomes, was evaluated in 50 specific-pathogen-free kittens after experimental challenge exposure with feline infectious peritonitis virus (FIPV). Ribavirin was administered daily for 10 to 14 days at 16.5 mg kg-1 bodyweight given per os, intramuscularly or intravenously beginning 18 hours after kittens were challenge exposed with FIPV. All kittens, including ribavirin-treated and untreated kittens, succumbed to FIP. Clinical signs of disease were more severe in the ribavirin-treated kittens and their mean survival times were shortened. The clinical efficacy of free ribavirin given intravenously at a reduced dosage (5.5 mg kg-1 bodyweight) was compared to that of ribavirin incorporated into lecithin containing liposomes (5 mg kg-1) intravenously. Drugs were given once daily for three consecutive days of each week for three weeks, beginning 18 hours after virus challenge exposure. There was no significant difference either in survival rate or severity of disease between kittens given free ribavirin, liposomal ribavirin or saline only. Because of its intrinsic toxicity and low therapeutic index against FIPV and its marginal antiviral activities in vivo at maximal doses, ribavirin cannot presently be recommended as primary antiviral chemotherapy against FIP. PMID- 8235083 TI - Scrapie associated fibril detection on decomposed and fixed ovine brain material. AB - Samples of cerebral cortex from eight scrapie affected sheep and two unaffected control sheep were stored for up to nine days at temperatures ranging from 18 degrees C to 29 degrees C. Scrapie associated fibrils (SAF) could be detected in proteinase K treated brain extracts from all the eight scrapie affected animals after five days storage and in six out of the eight after nine days storage. SAF could not be detected in any brain extracts from the two control animals. Formol saline fixed brain material from a further six scrapie affected and two clinically normal sheep, were also subjected to an extraction technique used to detect fibrils. No characteristic SAF were observed in any of these fixed samples. Long filamentous structures were observed in four of the fixed scrapie affected brain extracts and in one of the fixed unaffected control brain extracts. PMID- 8235084 TI - Serological characterisation of Actinobacillus pleuropneumoniae strains of serotypes 1, 9 and 11. AB - Various serological tests such as agglutination, coagglutination, indirect haemagglutination, immunodiffusion and counterimmunoelectrophoresis were used to characterise serologically Actinobacillus pleuropneumoniae isolates of serotypes 1, 9 and 11 using rabbit hyperimmune sera against serotypes 1 to 12. The rapid slide agglutination test using whole-cell suspension and the indirect haemagglutination test using whole-cell saline extract as antigens gave type specific reactions for serotypes 1 and 9. Antigens comprising saline extracts of boiled or autoclaved cells demonstrated common epitopes among the isolates of all three serotypes in the indirect haemagglutination test. Quantification of the type and group-specific antigens by the coagglutination test. Quantification of the type and group-specific antigens by the coagglutination test distinguished serotypes 1 and 9 strains from those of serotype 11. Results of absorption studies in immunodiffusion tests indicated that the cross-reactivity encountered among strains of serotypes 1, 9 and 11 might be due to common epitopes associated with cell-wall antigens. However, they certainly also have type-specific epitopes, possibly associated with superficially located, capsular antigens. PMID- 8235085 TI - Postnatal closure of the ductus arteriosus in West African dwarf goats. AB - Fifteen West African dwarf goats aged between 0 and 42 days were used to study the closure time of the ductus arteriosus using dye injection and histological techniques. The ductus arteriosus appeared haemodynamically patent in kid goats 0 to three days old. There was a progressive contraction of the smooth muscle cells of the wall of the ductus from birth, resulting in the obliteration of the lumen in four and six days at the pulmonary and aortic ends, respectively. This contraction phase was followed by disruption of the luminal endothelium, intimal thickening and necrosis of smooth muscle cells in goats nine to 18 days old. Beyond 21 days there was connective tissue formation resulting in the permanent sealing of the ductal lumen in goats 28 to 33 days old. Although lumina of varying sizes were observed in histological sections of the ductuses of four- to 32-day-old goats, these were haemodynamically insignificant. The results suggest three phases in the closure of the ductus arteriosus of West African dwarf goats which was completed between 28 and 33 days. PMID- 8235086 TI - Pulsed wave Doppler echocardiographic evaluation of intracardiac blood flow in normal sheep. AB - The blood flow velocities and flow patterns were evaluated in normal sheep across the four cardiac valves using pulsed wave spectral Doppler echocardiography. Merino ewes (n = 20) aged between 16 and 61 months with bodyweights ranging between 51 to 72 kg were evaluated. The sheep were fully conscious and required no sedation. The mean peak velocities for the left atrioventricular, right atrioventricular, aortic and pulmonary valves were 68.4, 51.9, 99.1 and 92.7 cm s 1, respectively. Additionally the wave forms were described and mean velocities and a variety of flow variables determined for each of these flow areas. The mean left atrioventricular valve E:A ratio was under 1 which differs from humans and dogs. Aortic and pulmonary flow variables were compared to each other. PMID- 8235087 TI - Association between cessation of leptospiruria in cattle and urinary antibody levels. AB - The shedding of Leptospira interrogans serovar hardjo in the urine of cattle and the local and systemic response to these organisms was monitored in experimentally and naturally infected animals. Twenty yearling heifers, 10 infected by the instillation of leptospires into the conjunctival sac (supraconjunctival route) and 10 infected intrauterinely, shed leptospires for up to 60 weeks after infection. Five of 15 naturally infected pregnant heifers with microscopic agglutination test titres > or = 1:300 continued to shed leptospires from 28 to 40 weeks after initial detection. Serovar hardjo was isolated infrequently from the urine of a further five naturally infected animals during the first eight weeks of the study but no leptospires were isolated from the remaining five animals for the duration of the study period (84 weeks). Cessation of leptospiruria in the 20 experimentally infected animals and in the five naturally infected persistent excretors was invariably associated with a sharp increase in urinary anti-leptospiral IgG and IgA antibody levels. PMID- 8235088 TI - Characterisation of Ostertagia ostertagi antigens by the different bovine immunoglobulin isotypes. AB - Antigenic differences between the developmental stages of Ostertagia ostertagi were studied by SDS-PAGE and Western blotting. Gel electrophoresis showed a complex protein pattern different for every stage with the O ostertagi fourth stage larvae (L4) showing an intermediate protein pattern between the third stage larvae (L3) and the adult stage. Immunoblotting showed that IgG1, IgG2 and IgM immunoglobulins present in serum from uninfected calves identified several O ostertagi antigens at every stage. When using serum from O ostertagi infected calves, O ostertagi specific IgG1 was the predominant bovine immunoglobulin. Specific IgG2 and IgM responses were also observed, while specific IgA antibodies were hardly detectable. Severe IgG1 cross reactivity was demonstrated when using anti-Cooperia oncophora serum. PMID- 8235089 TI - Influence of Newcastle disease virus on the severity of Pasteurella anatipestifer infection in turkeys. AB - This study was designed to examine whether vaccine or virulent strains of Newcastle disease virus (NDV) would potentiate the disease caused by Pasteurella anatipestifer infection in turkeys. The studies were conducted in turkeys of two age groups. There were three experiments. In two experiments four-week-old turkeys were exposed either to vaccine or virulent strains of NDV after experimental P anatipestifer infection. In the third experiment 14-week-old turkeys were first exposed to virulent NDV and superimposed with P anatipestifer infection. In experiment 1, one bird died where P anatipestifer was given in combination with the vaccine strain of NDV. However, there was no difference in the clinical signs, gross lesions and histopathology compared with turkeys given P anatipestifer alone. In experiment 3 where turkeys received a virulent strain of NDV in combination with P anatipestifer, birds became dyspnoeic and showed signs of illness. There was a difference in the course of the disease, gross lesions and histopathology when compared with turkeys that received P anatipestifer only. PMID- 8235090 TI - Histochemical and morphometric properties of muscles of the upper airway of goats. AB - This study investigated the histochemical and morphometric properties of fibres in laryngeal, hyoid, tongue and pharyngeal muscles which contribute in maintaining patency of the upper airway. Muscle specimens from adult female goats were stained for nicotinamide adenine dinucleotide dehydrogenase-tetrazolium reductase and myosin adenosine triphosphatase activities, and the composition and size of the fibre types determined. These muscles contained types 1, 2A, 2B and 2C fibres with type 2 fibres predominating and the fibres possessed oxidative enzyme activity suggesting fast contraction speed and yet moderate resistance to fatigue. Abductor laryngeal muscles contained more type 1 fibres than the adductors. Among pharyngeal muscles fibre size and type 1 fibre composition increased progressively from the hyopharyngeus caudally. Upper airway muscles contained relatively small fibres (range of mean diameter: 25.7 to 46.1 microns) with the pharyngeal and lingualis proprius muscles containing the smallest fibres. These properties might influence the response of upper airway muscles to neuromuscular blocking drugs. PMID- 8235091 TI - Effect of thiomolybdate and ammonium molybdate in pregnant guinea pigs and their offspring. AB - Groups of eight guinea pigs and their offspring were given drinking water containing molybdenum as ammonium molybdate (AM) or thiomolybdate (TM) throughout and subsequent to pregnancy. All adult females had oestrous cycles and conception rates were unaffected. Fetal death was common in groups given the high dose of TM. The concentration of copper in liver was reduced in all groups at all ages except for pups killed at birth from animals given AM. The concentration of molybdenum was elevated in liver and kidney of all groups and was statistically significant in the majority. The concentration in plasma of copper, molybdenum and copper insoluble in trichloroacetic acid was elevated in all groups. Superoxide dismutase activity was significantly reduced in dams and six-week-old pups in which TM administration commenced before mating. Histological damage occurred in the pancreas of animals given AM or TM. The effects on the fetus and pancreas were considered to result from copper deficiency rather than molybdenum toxicity. PMID- 8235092 TI - Folate sensitive common fragile sites in chromosomes of the domestic pig (Sus scrofa). AB - Chromosomal fragile sites were studied in blood lymphocytes of the pig after induction with 0, 5, 10 and 20 micrograms of fluorodeoxyuridine (FudR) ml-1 of culture. The percentage of cells showing fragile sites and total breaks and gaps in 100 cells was significantly higher in all treated cultures than the control (P < 0.001). A concentration of 5 to 10 micrograms of FudR ml-1 of medium is recommended as the optimum for mitotic index and fragile site expression in the pig. Localisation of the fragile sites was by GTG banding after destaining of the conventionally stained Giemsa preparations. Common fragile sites in the pig were identified in 24 sites on 10 chromosomes. Some fragile sites corresponded to the breakpoints in known reciprocal translocations. PMID- 8235093 TI - Behavioural responses of lambs of three ages in the first three hours after three methods of castration and tail docking. AB - The behavioural responses of groups of seven lambs were compared with control groups after castration and tail docking by rubber rings, application of a Burdizzo clamp in addition to a rubber ring and after surgical castration at five, 21 and 42 days. All methods at all ages produced changes in behaviour which were interpreted as indicative of considerable pain. The rubber ring groups showed most changes in behaviour at all ages. The rubber ring with Burdizzo groups showed least changes and some lambs in these groups showed much less response than others. The surgical groups showed some behavioural responses which were different, both qualitatively and quantitatively to those in other groups. It is concluded that indices used for recognition and assessment of acute pain received conditional support, that modification of the rubber ring with Burdizzo may provide the least painful method without local anaesthesia and that age had little effect on the responses. PMID- 8235094 TI - Changes in plasma cortisol concentration in lambs of three ages after three methods of castration and tail docking. AB - Lambs were handled only or castrated and tail docked at five, 21 and 42 days of age by either surgery, rubber ring or rubber ring and Burdizzo. Plasma cortisol was measured in blood samples taken before and at 12, 24, 36, 48, 60, 72, 84, 96, 138 and 180 minutes after castration and docking. Pre-treatment and peak cortisol values were highest in five-day-old lambs. The peak cortisol values, at each age, were similar for surgery and rubber ring groups. However, the peak occurred earlier after surgery and rubber ring Burdizzo than after rubber ring only treatment. The cortisol peak was 28 nmol litre-1 lower after rubber ring Burdizzo than surgery or rubber ring only. Plasma cortisol returned to pretreatment values within 84 minutes after rubber ring Burdizzo, 96 to 138 minutes after rubber ring only but not within 180 minutes after surgery. The changes in plasma cortisol together with the changes in behaviour suggest that the rubber ring Burdizzo method of castration and docking of lambs at all ages, was probably the least painful of the methods tested. PMID- 8235095 TI - Erythrocyte ALA-d activity in experimentally lead-poisoned ducks and its change during treatment with disodium calcium EDTA. AB - To determine useful procedures for the diagnosis and prognosis of lead poisoning in waterfowl caused by ingestion of lead pellets, erythrocyte delta aminolevulinic acid dehydratase (ALA-d) was investigated in experimentally lead poisoned ducks. A highly positive correlation was observed between the concentration of blood lead and the ALA-d activity ratio (the ratio of activated:non-activated enzyme activity) in those birds given seven lead pellets (3 mm diameter). The ALA-d activity ratio rapidly increased after the administration of lead pellets, but began to fall immediately after the initiation of disodium calcium ethylenediamine tetra-acetate (CaEDTA) therapy which resulted in a rapid decrease in the concentration of lead in the blood of these birds. In contrast, the ALA-d activity remained inhibited even after blood lead levels began to decrease following treatment. These results demonstrated that the ALA-d activity ratio is a very useful and sensitive indicator for the diagnosis and evaluation of therapeutic effects after lead poisoning in waterfowl. PMID- 8235096 TI - In vitro infection of bovine brain endothelial cells by Cowdria ruminantium. AB - Endothelial cells from bovine brain capillaries (BBEC) were successfully infected with the Gardel stock of Cowdria ruminantium. Growth conditions of cowdria in BBEC and bovine umbilical endothelial cells (BUE) were identical with average intervals between passages of 8.6 and 8.9 days in BBEC (17 passages) and BUE (28 passages), respectively. The time required to complete the life cycle and the morphology of the parasite were identical in both types of cells. The demonstration that BBEC may be infected with C ruminantium offers the means to study the mechanism of infection of the blood brain barrier endothelium by this parasite. PMID- 8235097 TI - Twenty-four hour gastric pH monitoring and blood gastrin concentrations in fasted ponies. AB - Gastric pH varied widely among eight fasted ponies, of which seven exhibited discrete episodes of spontaneous alkalinisation (SA). SA occurred at all times of the study and no significant variation in the summary variables of pH (median pH, mean pH and percentage of readings exceeding pH 4.0) was noted among the periods 0 to eight, eight to 16 and 16 to 24 hours. The occurrence of SA has significant implications for the performance of acid secretory studies in the fasted pony. There was, however, no significant correlation between pH and plasma gastrin concentration measured using a commercially available radioimmunoassay kit validated for use in the horse. This implies a lack of dependency of the incidence of SA on plasma gastrin or, conversely, of plasma gastrin on the variable pH associated with SA. PMID- 8235098 TI - Use of digitized fluorescein angiogram system to evaluate laser treatment for subretinal neovascularization: technique. AB - In patients with subretinal neovascular membranes (SRNVM), persistence of the SRNVM after laser treatment is a major cause of decreased visual acuity. Persistence is associated with inadequate initial laser treatment. A technique using a digitized fluorescein angiogram (DFA) system is presented that allows immediate evaluation of the adequacy of laser treatment for SRNVM. This involves overlaying a red-free photograph taken after treatment on a fluorescein angiogram taken before treatment using a digital angiogram system that evaluates the adequacy of treatment. A protocol for augmentation of laser treatment has been developed that will aid in the reduction of persistence after laser treatment of SRNVM. PMID- 8235099 TI - Evaluation of persistence of subretinal neovascular membranes using digitized angiographic analysis. AB - A study was conducted using the digitized fluorescein angiogram (DFA) system to evaluate adequacy of coverage of laser treatment for subretinal neovascular membranes (SRNVM) and to confirm that complete coverage lowers the incidence of persistent SRNVMs. Ten consecutive cases were evaluated with the DFA system after undergoing a single laser treatment. Three cases were judged to be adequately treated, and the remaining seven cases were judged to be inadequately treated. The DFA system was then used during photocoagulation in 16 consecutive cases to verify complete coverage of the SRNVM or to indicate the need for photocoagulation augmentation. Despite augmentation, 2 of the 16 cases remained inadequately treated. Overall, the cases that were adequately treated had a 12% (2/17) persistence rate, whereas those that were inadequately treated had a persistence rate of 67% (6/9). The data indicate that adequate coverage of SRNVMs is essential to prevent persistence. PMID- 8235100 TI - Ocular findings in neonates after extracorporeal membrane oxygenation. AB - Extracorporeal membrane oxygenation (ECMO) is a modified, prolonged cardiopulmonary bypass procedure used to treat newborns who have reversible cardiac or respiratory failure. The venoarterial bypass technique requires cannulation of both the right carotid artery and the internal jugular vein, and after decannulation these vessels are permanently ligated. Left-sided retinal vascular changes after ECMO have been reported, and were attributed to ligation of these vessels. A retrospective review of the results of ocular examinations of 86 infants who had undergone ECMO therapy at Childrens Hospital in Los Angeles between March, 1987 and May, 1991 was conducted. Normal findings were noted in 73 infants. One infant had bilateral retinal vascular tortuosity, and 12 infants had incidental ocular findings, but there was no evidence of left-sided retinal hemorrhage, venous congestion, or tortuosity. Our results suggest that left-sided retinal vascular changes after ECMO do not occur, occur only rarely, or clear rapidly and result in no permanent retinal damage. To unequivocally rule out the possibility that ECMO can cause transitory retinal vascular changes, we recommend further prospective studies, with ocular examinations performed before, during, and after ECMO. PMID- 8235101 TI - Visual outcome in acute retinal necrosis. AB - Acute retinal necrosis (ARN) is generally a progressive retinitis, often complicated by retinal detachment and a poor visual outcome. A series of 12 consecutive patients (13 eyes) with ARN is reported. All patients were examined early, with a limited extent of disease (in 77% of the eyes, less than 25% of retina involved). Patients were treated with high-dose intravenous acyclovir (10 mg/kg every 8 hours) and laser photocoagulation. During a follow-up period of 3 to 21 months, only 1 patient developed a rhegmatogenous retinal detachment resulting in visual loss. No patients developed bilateral disease once intravenous antiviral therapy was begun. Final visual acuity was 20/40 or better in 6 eyes (46%), and 20/400 or better in 12 eyes (92%). Two patients were examined within 6 weeks of extracapsular cataract extraction with intraocular lens implantation. In the majority of the cases, clinical findings were limited to several quadrants of the peripheral retina. Eyes with limited disease, if aggressively treated, have a relatively good visual prognosis. PMID- 8235102 TI - An animal model of focal, subacute, viral retinitis. AB - To study local intravitreal therapies for retinitis due to herpes viruses, an animal model of focal, subacute, relatively nonlethal herpes family retinitis is needed. Herpes simplex virus type 1 (HSV-1) was injected into the subretinal space of 33 Dutch pigmented rabbit eyes. The animals were observed for up to 42 days after the inoculation. All inoculated eyes developed a focal, enlarging area of retinitis in a predictable manner, showing focal enlarging areas of retinal opacification and necrosis with variable retinal hemorrhage. In the inoculated eyes, retinal detachment developed in all animals within 21 days; 33% of the animals developed focal retinitis in the uninoculated eye. Histologic examination showed encephalitis to be present in 11 (73%) of the 15 animals studied after 1 week. This model may be used to evaluate the therapeutic efficacy of new antiviral agents and modalities in the treatment of herpes family viral retinitis. The model is most similar to herpes simplex or zoster retinitis in humans, but also shares some similarities (and differences) with cytomegalovirus (CMV) retinitis in humans. PMID- 8235103 TI - Harvey Lincoff, noted ophthalmologist and surgeon, receives Man of the Year Award. PMID- 8235104 TI - A comparison of argon and diode photocoagulation combined with supplemental oxygen for the treatment of retinopathy of prematurity. AB - The efficacy of argon and diode laser photocoagulation of the avascular peripheral retina for threshold retinopathy of prematurity (ROP) was compared in a prospective trial. The study group included 9 premature infants (17 eyes). In each infant, one eye was treated with the diode laser indirect ophthalmoscope (LIO), and the fellow eye was treated with the argon LIO. Supplemental oxygen therapy was administered after laser treatment to all nine infants. The mean duration of the follow-up period was 9.7 +/- 2.6 months. All 17 eyes (100%) had complete regression of ROP and favorable outcomes. The diode and argon LIO appear to be equally effective in treating threshold ROP. Two patients sustained burns of the tunica vasculosa lentis and anterior lens capsule in the argon laser treated eye but not in the fellow diode treated eye. Photocoagulation with either the diode or argon LIO, combined with supplemental oxygen treatment, appears to be a promising treatment for retinopathy of prematurity. PMID- 8235105 TI - Visual abnormalities associated with high-energy microwave exposure. AB - A 44-year-old man was accidentally exposed to high-energy microwave irradiation. After resolution of facial erythema and iritis, he noted a foreign body sensation and blurring of vision. Ophthalmoscopic examination showed bilateral, small hard drusen. Ancillary tests were consistent with abnormal cone function. Electroretinogram testing revealed a marked decrease in the flicker electroretinogram. Results of D15 and Farnsworth Munsel Hue 100 color tests were abnormal. Two years later, the patient's visual acuity was stable at 20/25 in both eyes; however, results of flicker electroretinogram test remain markedly decreased. PMID- 8235106 TI - Electrophysiologic detection of choroidal detachment in eyes with rhegmatogenous retinal detachment. AB - The base values of electrooculograms (EOGs) of 39 patients with new, unilateral rhegmatogenous retinal detachment and clear media were studied. None of the patients had a history of ocular surgery. Ophthalmoscopic examinations revealed choroidal detachment in 11 eyes. The mean (+/- SD) amplitude, expressed as a percentage of the amplitude recorded for unaffected fellow eyes, was 116 +/- 31% in the 28 eyes without choroidal detachment, and 39 +/- 10% in the 11 eyes with choroidal detachment. This difference was statistically significant (P < 0.0001). There was no significant correlation in either group between baseline value and extent of retinal detachment or intraocular pressure. These results indicate that choroidal detachment associated with rhegmatogenous retinal detachment can be revealed electrophysiologically by the baseline value of an EOG. PMID- 8235107 TI - Fatal acute myocardial infarction after intravenous fluorescein angiography. AB - Since its introduction, fluorescein angiography has been widely used to investigate diseases of the ocular fundus. A case of fatal acute myocardial infarction after intravenous fluorescein angiography is presented. This appears to be the first case documented by autopsy in which the findings are compatible with myocardial infarction as the cause of death. Although there are no known contraindications for fluorescein angiography in patients with a history of cardiovascular disease, the indications for this elective procedure should be carefully reviewed in such patients. Adequate emergency resuscitation equipment should be available in the fluorescein angiography suite. PMID- 8235108 TI - Intraocular gnathostomiasis. AB - The clinical and histologic features of an intraocular parasite from a 29-year old woman are reported. The live organism was located in the posterior vitreous near the optic disc and was successfully removed by vitreous aspiration. The parasite was identified as an advanced stage larvae of Gnathostoma spinigerum. This is the first case report of the scanning electron microscopic findings of intraocular gnathostomiasis. The features of nine previously reported cases of intraocular gnathostomiasis are reviewed. PMID- 8235109 TI - Silicone oil as a delivery vehicle for BCNU in rabbit proliferative vitreoretinopathy. AB - The antiproliferative properties of a single intravitreous injection of BCNU [carmustine; 1,3-bis(2 chloroethyl)-1-nitrosourea] dissolved in 1 ml of medical grade, 1,000-centistoke silicone oil were evaluated in a rabbit model of proliferative vitreoretinopathy (PVR). 10 micrograms and 15 micrograms of BCNU in 1 ml silicone oil resulted in a 46% and 27% reduction in the incidence of retinal detachments, respectively. Chi-square tests showed a significant statistical difference (P < 0.05) between the control and treated groups. However, histopathologic examination of the eyes injected with BCNU and silicone oil indicated some retinal disorganization even at the lower therapeutic levels. With a therapeutic index of < 1.0, BCNU in silicone oil has a low margin of safety for treatment of PVR. Nevertheless, this study demonstrates that silicone oil when used for retinal tamponade in PVR may be the vehicle for delivery of a lipophilic antiproliferative agent. PMID- 8235110 TI - Indocyanine green-enhanced diode laser treatment of melanoma in a rabbit model. AB - Indocyanine green (ICG)-enhanced laser therapy was evaluated for the treatment of experimental intraocular melanoma. Immediately after an intravenous injection of ICG, a 790-nm chromophore, 810-nm semiconductor diode laser was used to irradiate Greene hamster melanomas that had been implanted in the iris of rabbits. ICG enhanced laser treatment of melanoma (14 eyes) was compared with treatment by laser alone (4 eyes), ICG alone (1 eye), and no treatment (2 eyes). Tumors treated with ICG-enhanced laser showed no growth after treatment, as judged by clinical examination and photography. Histologically, 4 of the 14 tumors treated with ICG-enhanced laser showed total necrosis, whereas the remaining 10 tumors treated similarly demonstrated only rare viable cells around blood vessels or at the tumor periphery. Laser treatment without ICG enhancement resulted in only superficial tumor necrosis, and all four of these tumors continued to grow after treatment. With further evaluation, indocyanine green in combination with a commercially available diode laser may be useful in the treatment of ocular melanoma. PMID- 8235111 TI - Bread crumb-flecked retinopathy. PMID- 8235113 TI - The Fight for Sight scientific grants program. PMID- 8235112 TI - Tie up retinal surgery with the silicone lace. PMID- 8235114 TI - [Transcatheter embolization of ascending aorto-pulmonary fistula combined with coronary arteriovenous fistula]. AB - We encountered a patient with a coronary arteriovenous fistula accompanied by an ascending aortopulmonary fistula, a rare malformation. The patient was a 42-year old female complaining of exertional dyspnea and chest pain at rest. She noticed dyspnea while going up and down the stairs, and chest pain at rest at the age of about 40 years. While she was being treated for hypertension, a cardiac murmur was detected. She was admitted to our hospital for close examination. Coronary angiography (CAG) showed a coronary arteriovenous fistula that branches from the left anterior descending branch and forms an aneurysm, running to the trunk of the pulmonary artery. On March 28, 1989, ligation of the fistula was performed. Postoperative CAG demonstrated an ascending aorto-pulmonary fistula that flowed to an aneurysm adjacent to the pulmonary artery. On November 29, transcatheter embolization was performed. She was discharged 7 days after embolization without complications. She is now being treated on an outpatient basis, and the symptoms in her chest have disappeared. Transcatheter embolization used in this patient may also be applicable in the treatment for coronary arteriovenous fistulae. PMID- 8235115 TI - [A case of congenital antithrombin III deficiency with pulmonary thromboembolism and cerebellar infarction]. AB - A 41-year old woman was admitted to a local hospital because of leg pain, chest pain and dyspnea after taking estrogen for two months for irregular menstruation. On admission to the hospital, her lung scintigram showed multiple segmental perfusion defects in the right lung. Pulmonary angiography showed several thrombi in the proximal pulmonary artery. A diagnosis of pulmonary thromboembolism was made. AT-III activity was found to be only 50% of normal value, indicating that the pulmonary thromboemboli were due to AT-III deficiency. The patient was then put on anticoagulants for two years. When she was 43-year-old, she was admitted to Keio University Hospital because of worsening of dyspnea. After admission her dyspnea had got better due to bed rest. Her brain CT, which was performed because of her gait disturbance, indicated past right cerebellar infarction. We gave her 2 mg of warfarin and maintained a thrombotest of about 30%. The investigation of her family revealed that her son also showed decrease AT-III activity, indicating a congenital abnormality. 26 families with this disease have been reported in Japan. Most had venous thromboemboli, but only a few cases had brain infarction as well. In addition, it is suggested that an estrogen therapy for irregular menstruation may have contributed, in this particular case, to the onset of thromboemboli. PMID- 8235116 TI - [Two case reports of refractory heart failure treated by continuous ambulatory peritoneal dialysis]. AB - In this report, we present 2 cases of severe congestive heart failure and mild renal insufficiency in patients who underwent continuous ambulatory peritoneal dialysis (CAPD) after stabilization using the extracorporeal ultrafiltration method (ECUM). Long-term good control of congestive heart failure was achieved following the institution of CAPD. Case 1, a 58-year-old woman with rheumatic arthritis and diabetes mellitus had anteroseptal myocardial infarction at the age of 52. And case 2, a 68-year-old man, who underwent coronary artery bypass surgery at the age of 66 and had extensive anterior infarction after the operation. They were admitted to the hospital with dyspnea due to congestive heart failure. In both cases, systolic cardiac function was severely impaired and mild renal insufficiency was present at the time of hospitalization. After admission, symptomatic relief was not obtained by conventional therapies and symptoms of congestive heart failure worsened until the patients suffered from severe respiratory distress even at rest. ECUM was then instituted to remove excess fluid and clinical improvement was achieved. After the initiation of ECUM, responsiveness to diuretics was not restored, and the procedure was necessary every day or every other day for the prevention of symptoms due to fluid overload. About 20 days after the initiation of ECUM, CAPD was begun for the long term control of congestive heart failure and renal failure, and for the purpose of hospital discharge. Good control of heart failure was achieved after the initiation of CAPD.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235117 TI - [Cardiovascular diseases and hemorheology]. PMID- 8235118 TI - [Drug-induced pulmonary disorders]. PMID- 8235119 TI - [Evaluation of intra- and inter-observer reliability]. PMID- 8235120 TI - [Eosinophils: eosinophil's infiltrations into the lung tissue and increases in peripheral blood]. PMID- 8235122 TI - [Intravascular ultrasound]. PMID- 8235121 TI - [Nasal CPAP and nasal BiPAP]. PMID- 8235123 TI - [An experimental study of catheter ablation using microwave energy via coaxial electrode catheter]. AB - The purpose of this study was to examine the efficacy, arrhythmogenicity, and the size of ablated lesions of catheter ablation with microwave energy. Microwave energy generated by a device with a frequency of 2450 MHz, was delivered via a bipolar electrode coaxial catheter with an electrode ball tip. Microwave ablation was applied (50 watts for 15-150 sec.) on the left ventricular endocardium in 6 dogs. No ventricular arrhythmia occurred during ablation in any of the dogs when microwave catheter ablation was applied for less than 45 sec. However, when the duration of microwave catheter ablation was longer than 45-60 sec, ventricular premature contractions were observed in all 6 dogs. When ablation time was set for a longer time, the dogs developed nonsustained ventricular tachycardias. Nevertheless, ventricular arrhythmias after ablation did not occur in all dogs. Ventricular programmed stimulation after ablation did not induce ventricular tachycardia in any of the dogs. When ablation time lasted more than 120 sec, the coagulation layer was extended to the epicardium in all 6 dogs. The results of this study suggest that microwave ablation is feasible for the treatment of tachyarrhythmias from a deep focus of the ventricular myocardium. PMID- 8235124 TI - [Frequency dependent prolongation of inter-atrial conduction time by mexiletine in human atrium]. AB - To investigate frequency dependent conduction slowing by mexiletine in the human atrium, we examined inter-atrial conduction time (IACT) at different stimulation frequencies (100/min to 220/min) in 13 patients with paroxysmal atrial fibrillation (Paf) and 7 patients without Paf. IACT was prolonged as the stimulation frequency was increased, and either before or after mexiletine administration IACT was longer in the Paf group (max 114 +/- 9 msec, p < 0.01, mean +/- SD) at any stimulation frequency than in the non-Paf group (max 100 +/- 8 msec). The change in IACT induced by mexiletine administration (% delta IACT) was larger in the Paf group (max 14.8 +/- 5.4%, p < 0.05) than in the non-Paf group (max 5.5 +/- 2.2%) at stimulation frequency over 140/min. Effective refractory period measured at the high right atrium was slightly decreased due to mexiletine in both the non-Paf and the Paf group. In conclusion, mexiletine showed frequency dependent suppression of conduction in the human atrial myocardium especially in patients with Paf. PMID- 8235125 TI - [Application of lactate threshold in the exercise program for patients with coronary heart disease]. AB - This study was conducted 1) to measure the exercise intensity corresponding to lactate threshold (LT) and 2) to clarify the method of prescribing LT level for patients with coronary heart disease (CHD) on the assumption that LT level indicates an optimal exercise intensity for those patients. Three males and 9 females with CHD, aged 57.6 +/- 7.7, participated in this study. Oxygen uptake (VO2), heart rate (HR), and ratings of perceived exertion (RPE) at LT and at maximal exercise were measured before (pre), 4 months (mid) and 8 months (post) after exercise conditioning. The intensity of most exercises was maintained at the level of LT. A significant increase was found in VO2LT from pre- to mid-test, and further increase was found from mid- to post-test after matching exercise intensity to the improvement in VO2LT. It is suggested that matching the progression of the absolute exercise intensity with the training-induced improvements of aerobic capacity i.e., maintaining the exercise intensity at LT for a sustained period of time, is a valid procedure for further improvement in aerobic capacity of CHD patients. PMID- 8235126 TI - Factors affecting distribution of airflow in a human tracheobronchial cast. AB - Air velocity was measured at end airways of hollow replicate casts of the human tracheobronchial tree in order to determine the flow distribution within casts extending to 3 mm diameter airways. Measurements were made by hot-wire anemometry for constant inspiratory flow rates of 7.5, 15, 30 and 60 L.min-1. Average flow distribution among the lung lobes was as follows: right upper, 18.5%; right middle, 9.2%; right lower, 32.3%; left upper, 15.7%; and left lower, 24.3%. An empirical model derived from the experimental flow distribution data demonstrated the effect of various morphometric parameters of the hollow cast on the distribution of airflow. Airway cross-sectional area, branching angle and total path-length were found to have the greatest influence. As the tracheal flow rate decreased from 60 to 7.5 L.min-1, the influence of branching angle was reduced, while total path-length became more influential. These results provide evidence for the transition of flow regimes within the TB tree within normal physiological flow ranges. PMID- 8235127 TI - Independent modulation of horse airway smooth muscle by epithelium and prostanoids. AB - The effects of epithelial removal and cyclooxygenase inhibition on contractions induced by exogenous acetylcholine (ACh) and electrical field stimulation (EFS) were evaluated in horse tracheal strips and bronchial rings. Epithelial removal potentiated the response to ACh but had no influence on the response to EFS. The effect of epithelial removal was not altered by pretreating the tissues with meclofenamate, a cyclooxygenase inhibitor. In trachealis strips, meclofenamate augmented contractions induced by EFS but not by ACh. In bronchial rings, meclofenamate augmented EFS-induced contraction to a greater extent than ACh induced contraction. These effects of meclofenamate were epithelium-independent. We conclude that horse airway epithelium produces a relaxant factor that is not a prostanoid. Endogenous prostanoids originating from non-epithelial sites inhibit only cholinergic nerves in the trachea but both parasympathetic nerves and smooth muscle in the bronchi. PMID- 8235128 TI - Abdominal muscle activation by expiratory threshold loading in awake dogs. AB - Abdominal muscle activation produced by expiratory threshold loading (ETL) helps prevent an increase in FRC thus, optimizing diaphragm length and defending VT. However, anesthesia may affect abdominal muscle activation, and the pattern of recruitment and level of activation of individual abdominal muscles may well be dependent on body position. Therefore, individual abdominal muscle response to ETL was assessed in awake dogs, lying in the lateral decubitus position. Eight, tracheotomized dogs were chronically instrumented with sonomicrometer transducers and bipolar, fine wire EMG electrodes, in each of the four abdominal muscles. ETL produced increases in active, expiratory shortening of the transversus abdominis (TA), internal oblique (IO) and external oblique (EO). In addition, tonic activity, assessed from a decrease in baseline length, increased in the IO. There was a significant increase in FRC during ETL but it was less than would be expected without tonic and phasic abdominal muscle activation. Although FRC increased, VT and breathing frequency were maintained. As was found previously in supine, anesthetized dogs, the internal abdominal muscle layer (TA and IO) was recruited preferentially; substantiating its greater role in the defence of lung volume. PMID- 8235129 TI - The pattern of respiration with increasing metabolism in a small dasyurid marsupial. AB - Previous studies have indicated that the pattern of respiration in marsupial and placental mammals may be different. Some marsupials have larger tidal volumes and slower respiratory rates under basal conditions. This study examined the respiratory responses of a small marsupial, Dasyuroides byrnei, to increasing metabolic demand. The highest metabolic rate elicited by cold exposure in a helium-oxygen atmosphere was 10.4 times the basal metabolic rate. Basal tidal volumes and respiratory rates were 138% and 46% respectively of the values predicted for placental mammals. The increasing oxygen demands of metabolism were met by increases in ventilation rather than by changes in oxygen extraction. Initially, tidal volume increased until it reached a maximum value 2.6 times that of basal tidal volume. Subsequently, ventilation was augmented by an increase in respiratory frequency. Ventilatory accommodation to an increasing oxygen demand indicated that D. byrnei has an excellent respiratory capacity to deal with the thermogenic demands of a cold environment. PMID- 8235130 TI - Depth profiles of pH and PO2 in the isolated brain stem-spinal cord of the neonatal rat. AB - We have measured depth profiles of extracellular pH (pHECR) and PO2 (PtO2) as well as the kinetics of changes of pHECR in the isolated brain stem-spinal cord preparation of the neonatal rat using pH and PO2 microelectrodes that entered from the ventral surface. When the preparation was superfused with control mock cerebrospinal fluid (Control mock CSF; pH = 7.5, PO2 = 630 Torr, PCO2 = 28 Torr, at 27 degrees C), the pH in the medulla diminished with a nearly constant gradient from the surface to a depth of about 1000 microns, the slope being about 0.1 pH unit per 100 microns. A similar gradient in the 200 to 300 microns of the CSF above the surface suggested existence of unstirred layers despite continuously flowing superfusate. The pH gradient in the spinal cord was somewhat smaller than that in the medulla. The PO2 gradients in both medulla and spinal cord were about 100 Torr per 100 microns from 200 microns above to 100 to 200 microns below the surface; PO2 reached zero at about 450 (medulla) to 600 microns (spinal cord). Although the preparation was anoxic and acidic except for a small layer below the surface, respiratory activity was recorded for several hours in C4 phrenic roots. The kinetics of changes in pHECF were recorded at 100 and 200 microns depth while rapidly replacing the control mock CSF by more acidic CSF, either with increased PCO2 ("Respiratory acidosis") or by adding fixed acid ("Metabolic acidosis"). The changes in pHECF were smaller than those in pHCSF, particularly during respiratory acidosis, as a result of the buffering of the brain tissue. Our results show the importance of superficial layers of the ventral medulla in producing respiratory rhythmicity; they further suggest that somewhat alkaline CSF (pH about 7.8) should be used in this preparation to ensure physiologic surface pH values despite unstirred surface layers. PMID- 8235131 TI - Hypercapnia and medullary neurons in the isolated brain stem-spinal cord of the rat. AB - We have extracellularly recorded single neuron activity in the ventral medulla of the isolated brain stem-spinal cord preparation of the neonatal rat (37 preparations) in order to test their sensitivity to changes in CO2/H+. Search for neuronal activity was performed when the preparation was superfused with control mock CSF (equilibrated with 2% CO2, 90% O2 in N2; pH = 7.8 at 27 degrees C). Neurons, found down to about 500 microns from the surface, could be classified as R neurons when they showed rhythmic discharge in phase with phrenic activity, recorded from C4 ventral roots; or as Non-R neurons when they did not exhibit such phasic discharge. Among the 89 Non-R neurons, 20 responded to rapidly replacing the control CSF by hypercapnic CSF (8% CO2, 90% O2 in N2; pH = 7.2) with increased, 44 with reduced activity, while 25 did not respond to hypercapnia. Five Non-R neurons became phasic with respiration during hypercapnia. Of the 14 R neurons, 10 fired predominantly in expiration (R-E), 4 in inspiration (R-I). Only one R-E and two R-I neurons were excited by hypercapnia, the remaining were either inhibited or did not respond. Excited Non R and R neurons were mainly encountered in rostral parts of those areas in the ventral medulla that have been reported as chemosensitive. PMID- 8235132 TI - Temperature and central chemoreceptor drive to ventilation in toad (Bufo paracnemis). AB - The central chemoreceptor drive to ventilation was assessed in unanesthetized toads, Bufo paracnemis, exposed to three different temperatures: 15, 25 and 35 degrees C. The acid-base status of the fourth ventricle was manipulated by mock CSF perfusion. In additional experiments, arterial pH was varied by inspiration of hypercapnic gas mixtures. Ventilation was measured directly by pneumotachography and arterial blood samples were analyzed using electrodes for pH and PO2. Regardless of temperature, the ventilatory control of acid-base status was predominantly central. Moreover, an increase in temperature was accompanied by a proportional increase in the ventilatory response to chemoreceptor stimulation by either lowered mock CSF pH or hypercapnia. The alphastat hypothesis could not adequately account for the temperature effects on the ventilatory responses to hypercapnia or on air convection requirements in the toad. PMID- 8235133 TI - [Psychosocial support in pregnancy as a strategy to promote the newborn's health]. AB - In this project we developed a social support and health education intervention meant to answer whether or not a program of this kind, within the present medical system, can improve perinatal outcomes. A multicenter randomized controlled trial was conducted in four Latin American countries. In Mexico, the project was carried out in a third level obstetric hospital. Patients were screened between the 15th and 22nd week of gestation. Inclusion criteria were factors associated with high risk of low birthweight. A total of 620 women were recruited, randomized and followed up during pregnancy and up to the 40th day post partum. Women in the intervention group were visited four to six times during pregnancy, could use a telephone hot line and a consultation service. A poster and a pamphlet meant to reinforce health education were handed out in the first visit. A "guided tour" for women at the end of pregnancy was organized periodically. Outcomes were measured in both groups at the 36th week of gestation, post partum and 40 days after delivery. Neonatal outcomes were obtained by blind interviewers from the clinical records. Neither in the multicenter nor in the Mexican sample were we able to modify the incidence of low birthweight and prematurity. However, in Mexico we obtained positive effects on birthweight and gestational age in the whole sample and in some specific strata. These results strongly suggest the convenience of replicating the study in Mexico. PMID- 8235134 TI - [Surgery in portal hypertension. Which patient and which operation?]. AB - A prospective trial of a cohort of patients (N = 94) with portal hypertension and history of bleeding was selected for surgery based on strict clinical and laboratory criteria. All of them were treated with portal blood flow preserving procedures. The following selection criteria were used: good cardiopulmonary function without pulmonary hypertension and good liver function (Child-Pugh A). All patients were operated in an elective fashion and the operations performed were: selective shunts (N = 38) (distal splenorenal and splenocaval), low diameter mesocaval shunts (N = 13) and the esophagogastric devascularization with esophageal transection (Sugiura-Futagawa) (N = 43). Patients were selected for each operation according to their anatomical conditions. Sixty-one of the patients were cirrhotics. Operative mortality was 8% and rebleeding was observed in 5% of the cases. Postoperative encephalopathy was seen in seven patients (three selective shunts, two low diameter mesocaval shunts and two devascularizations). In 13 of 62 patients postoperatively evaluated by means of angiography, portal vein thrombosis was shown (seven selective shunts, two low diameter shunts and four devascularizations). Twenty-two patients with preoperative portal vein thrombosis (and treated with a Sugiura-Futagawa operation) were excluded from postoperative angiographic evaluation. Survival (Kaplan-Meier) was 85% at 60 months. Portal blood flow preserving procedures are the treatment of choice for patients with hemorrhagic portal hypertension and good liver function. The kind of operation is selected according to the individual anatomical status of the patient. PMID- 8235135 TI - [Surveillance of therapy with aminoglycosides at a specialized hospital]. AB - Aminoglycosides (AG) are excellent antibiotics against gram-negative bacilli infections, but their use implies potential ototoxicity and nephrotoxicity if an excessive dosage is prescribed. In this study we evaluated 105 episodes of AG treatment in 104 patients hospitalized in a specialty hospital in the city of Leon, Mexico. In each case a basal serum creatinine, weight and age were recorded. A single serum AG assay per episode of treatment was done. Either amikacin or gentamicin were one the AG prescribed. Prior to AG administration only 54 treatment episodes (51%) had a before treatment creatinine assay. The initial creatinine clearance showed normal values in 43% (23/54 episodes) and it was altered (below 80 mL/min) in 31 (57%) of the episodes. In the 31 cases with an altered renal function only 15 (48%) underwent dosage adjustment. In summary the AG were prescribed in excess in 54% (29/54) of the episodes. The serum AG levels within toxic range were observed in 11 episodes. These results suggest that in our hospital AG treatment is not adequately done and monitored. PMID- 8235136 TI - [Chemico-clinical permanent program of the NIH. I. The first phase of the program]. AB - BACKGROUND INFORMATION: This is the report of the first phase of an internal/external permanent program of the MNIH (Mexican National Institutes of Health). The permanent program evolved from a previous one operating from 1984 to 1988 in the MNIH (Refs 1-5). OBJECTIVE: To evaluate the precision of the assay systems used currently by the clinical chemistry laboratories of the MNIH. MATERIAL AND METHODS: A single batch of a lyophylized control serum prepared for the program by an agency of the Ministry of Health was used throughout the study. During a period of four months, the participants were provided with flasks of the control serum every two weeks to assay the analytes included in the routine daily work of each laboratory. They also received every two weeks an update of the mean of means and interlab CV of the information available. RESULTS: The 11 participants reported 6278 results of 24 analytes (6 enzymes, 8 electrolytes and 10 organic metabolites) assayed in 179 systems. The majority of the systems (162/179 = 91%) were autoanalyzers. There were 168 systems which could be evaluated in precision and 115 in accuracy. The rate of imprecision was 15% (25/168) and of inaccuracy 17% (19/115). CONCLUSIONS: 1. The participants were able to act as a reference group to assign the value of 12 analytes in a control serum with low concentration of most analytes. 2. Imprecision was concentrated in two participants (Labs B and J with 15 of the 25 imprecisions: see table 2). The second phase of the program will be to identify and if possible eliminate the sources of variation in the systems of participants B and J. PMID- 8235137 TI - [Current concepts on prolactin physiology: molecular variants and mechanisms of action]. AB - Prolactin is a polymorphic hormone with multiple biological functions. The versatility in the actions of prolactin depends on its structural polymorphism and perhaps on the feasibility of prolactin to be converted into different bioactive forms. In this review we attempt to summarize information concerning the synthesis, structure and mechanisms of action, including the heterogeneous nature of prolactin and its possible physiological significance in humans. PMID- 8235138 TI - [Structure and regulation of the expression of the angiotensinogen gene]. AB - Angiotensinogen is a glycoprotein synthesized mainly in hepatocytes and secreted into the circulation. Angiotensinogen is cleaved by the enzyme renin to produce angiotensin I, which is further converted into a vasoconstricting peptide, angiotensin II, the biologically active peptide of the renin angiotensin system. The concentration of angiotensinogen is rate-limiting in the production of angiotensin I and therefore plays an important role in the regulation of angiotensin II production. The development of recombinant DNA technology has introduced new directions for the study of the angiotensinogen molecule. The human, rat and mouse angiotensinogen gene contains five exons interrupted by four intervening sequences and spans 12 kb approximately. In its 5' flanking region multiple regulating elements, as well as the major control elements, are present. The cloning and sequencing of the angiotensinogen cDNA demonstrates the similarity of angiotensinogen to various serine protease inhibitors produced by the liver and was the beginning of studies looking for new physiological roles of angiotensinogen, in addition to the substrate for renin. The circulating levels of angiotensinogen are altered in many different physiological and pathological states. High levels of this protein are seen in hypercorticism, inflammation, pregnancy, and contraceptive therapy, and low levels are associated with adrenal insufficiency and converting enzyme inhibition. These variations are mostly due to modifications of the hepatic biosynthesis under the control of hormonal factors such as glucocorticoid, estrogen, thyroid hormone, insulin and angiotensin II. In addition, it has been found that these changes in the hepatic biosynthesis are due mainly to changes in the angiotensinogen gene expression. PMID- 8235139 TI - [Passive smoking, a problem of public health]. PMID- 8235140 TI - [Development of an immunoenzyme assay technique of ELISA type for detecting anti SSB antibodies. Comparative study of 2 kits]. AB - The anti-SSB antibodies were measured using two enzyme immunoassays (ELISA). The difference between the both results from the preparation of the SSB antigenic extract. The first method, developed in our laboratory, uses a purified SSB antigen extracted from calf thymus, while the other uses an antigen cloned by genetic engineering. We have realized an analytic investigation about the repetability, the reproducibility and the detection limit of our ELISA method and we have refined its evaluation in using a clinic study carried out on 203 subjects (55 had a Sjogren syndrome, 47 had a systemic lupus erythematosus, 17 had a rheumatoid arthritis, 13 a progressive systemic sclerosis, 10 a polymyositis and 61 healthy subjects (blood donors)). These measures were worked out in order to compare them with the Ouchterlony method of reference. The results we have obtained are totally similar to the ELISA methods, with a global correlation factor of 0.96 in spite of the difference on the preparation of SSB antigenic extract. The enzyme immunoassay is a lot more sensitive than the Ouchterlony method since, for a Sjogren sample, we obtain a sensitivity of 0.69 while the sensitivity is only of 0.51 for the immunoprecipitation. In the lupus sample, the sensitivity is respectively of 0.42 and 0.25. PMID- 8235141 TI - [Vertebral compression in epilepsy. Analysis of 8 cases]. AB - Epileptic seizures can produce dislocations or fractures of the limbs and vertebral compression fractures. We tried to determine the clinical and radiological features of the latter in 8 patients: 6 men and 2 women including one aged only 41 years. The often multiple compression fractures involved the first thoracic (n = 3) and the intermediate thoracic (n = 3) or lower lumbar (n = 2) vertebrae but, contrary to common osteoporosis, never the thoracolumbar hinge. In 2 cases, the appearance of the fracture and the neurological signs led to surgery. Posterior dislocation of the shoulder was associated with the fractures in 2 cases. Chronic alcoholism (4 cases), anti-epileptic drugs (1 case) or corticosteroids (1 case) might have facilitated a demineralization which was however not obvious on X-ray films. The seizure was a first ever event in 6 cases. In the absence of controls the fracture was considered to be spontaneous. These particularities explain why a metastatic compression fracture was initially suspected in 3 cases, leading to explorations which sometimes were aggressive. In the presence of a suspicious vertebral compression fracture the clinician must consider the possibility of an epileptic seizure, a trauma which is sufficient to explain the fracture, as shown by the complications of electro-convulsive therapy. PMID- 8235142 TI - [Medicalization of information systems in Internal Medicine and Geriatrics: a true medical challenge]. AB - Elderly people constitute an ever growing part of short-stay hospital patients. At the moment, 25% of these patients age aged 75 or more. The demographic curve, the effect of public health policies favouring treatment at home and the new characteristics of demand for care by the oldest patients will undoubtedly result in an increasing number of these hospitalizations. This tendency has repercussions on the organization of hospital structures, but it also raises the problems of specificity of consumption of goods and medical hospital units by elderly patients and of their financing as part of budget allotments. The progressive but imperative installation, forced on us by decrees, of medicalized management tools (e.g. the Information System Medicalization Programme) in public hospitals and some private clinics throws doubt on the value of this tool to describe the stays of elderly people. PMID- 8235143 TI - [Tick-borne diseases]. AB - Due to their worldwide distribution, from hottest to coldest climates, and due to their behaviour, ticks are capable of transmitting numerous human and animal bacterial viral or parasitous diseases. Depending on the disease, they play the role of biological vector or intermediate host. In France, six tick borne diseases are of epidemiologic importance. Q fever (not often tick-borne), Mediterranean Spotted Fever, Lyme disease, Turalemia (human and animal), Babesiosis and Tick-borne Viral Encephalitis. PMID- 8235144 TI - [Current aspects of brucellosis]. AB - In France, human brucellosis decreased significantly between the 70's (800 annual cases) and 1992 (102 cases), the result of the veterinary fight against animal disease. Bovin brucellosis is almost eradicated except into Massif Central, Pyrenees and Alpes, where goats and sheep are always responsible for the contamination of cows. B. melitensis 9/11 biovar 3 is the more frequent contaminant. Clinical features of human disease are unchanged but skeletal and visceral focalisations become less frequent than previously--Blood cultures--too often left--and seroagglutination remain classical means of diagnosis. I.I.F. and ELISA are more precise because recognition of specific IgM and IgA which are good evidence of recent or persistent infection. In vitro antibody production--assay (IVAP) is more sophistical; its value should be assessed. Today antibiotherapy for acute or subacute forms is based on associations: Tetracycline (2 g/d) or better Doxycycline (200 mg once daily for 45 d) are always associated with either Streptomycin (1 g daily for 21 days) or Rifampicin (900 mg once daily for 45 d). The first regimen is probably more efficient than the second one because relapses occur more often (5-10%) with Doxycycline-Rifampicin than with Doxycycline Streptomycin (relapses: 4-5%). In pregnant women Cotrimoxazole-Rifampin is efficient enough and atoxic if the treatment is stopped 7 days before delivery. The same regimen is recommended in children less 8 years of age. Veterinary action should be going on in spite of its cost. Individual measures (gloves, antiseptic solution for shoes, boiled milk...) are always necessary. Specific immunization (vaccin Merieux, France) often seems efficient despite a few light adverse effects; its use is recommended in farmers, veterinary and bacteriologic workers. PMID- 8235145 TI - [Infectious diseases transmitted by animal bites]. AB - The animal population, and most of all pets, entail a high risk of bites, some of them severe, which may lead to complications among which infection is a major one. Epidemiological data about the germs liable to grow (Pasteurellae, pyogenic germs, anareobes...) are helpful to guide curative, and even more, prophylactic approaches to treatment. PMID- 8235146 TI - [Horton's disease disclosed by involvement of the lower limbs]. AB - We report the case of a 62-year old woman hospitalized for acute ischaemia of the right lower limb, caused by stenosis of the superficial femoral artery associated with thrombosis of the deep femoral artery. Thromboendarterectomy was performed, and histology of the operative specimen showed thickening of the media with clusters of giant cells and fragmentation of the internal elastic lamina, without atheroma. The diagnostic of giant cell arteritis was then considered and confirmed by the presence of headaches, 38 degrees C fever and inflammatory syndrome with ESR at 75 mm in the first hour. On the other hand, biopsy of a temporal artery was negative. Corticosteroid therapy was prescribed and gave excellent clinical, laboratory and arteriographic results. It has now been demonstrated that the arteries of the lower limbs may be involved in Horton's giant cell arteritis, which is often overlooked. This involvement is rare and exceptionally proven at histology. Clinically, the most frequently described form is one of pseudo-arteritis with claudication, but a few cases of gangrene have been reported. Withdrawal of corticosteroid might be a facilitating factor. As in our patient, the arteriographic lesions are often segmental, multifocal and symmetrical, predominant in the superficial femoral artery and the arteries of the legs. The lesions regress under corticosteroid therapy, and surgery can usually be avoided. Anticoagulants are commonly prescribed when the large vessels are involved. When surgery is not indicated the diagnosis can be confirmed by biopsy of the temporal artery, which is positive in the majority of cases. PMID- 8235147 TI - [Remission under treatment of cutaneous xanthomatosis disclosing myeloma: apropos of a case with a three-year course]. AB - We report a case of IgG lambda multiple myeloma with inaugural cutaneous xanthomatosis. A three years follow up showed parallel evolution of xanthomatosis and monoclonal gammapathy during therapy suggesting that the association is not coincidental. A decrease in IDL catabolism could be responsible. The monoclonal gammapathy could react with IDL and disturb the recognition of apolipoprotein E by its hepatic receptor. PMID- 8235148 TI - [Bronchiectasis following colectomy for hemorrhagic rectocolitis]. AB - Pulmonary disease is an uncommon extraintestinal manifestation of inflammatory bowel disease. We report the case of a patient in whom colectomy for ulcerative colitis was followed by development of bronchiectasis. A discussion of the relation between ulcerative colitis and bronchial disease is presented. PMID- 8235149 TI - [Arteriopathies as occupational diseases]. AB - Arteriopathies of occupational diseases predominate in the upper limbs and are usually expressed as Raynaud's phenomenon ("white finger"), sometimes complicated by digital gangrene. Hand-arm vibration disease, hypothenar hammer syndrome, exposure to silica even without silicosis and vinyl chloride intoxication are the principal causes of occupational diseases acknowledged as such by the laws. In a personal series of 60 patients with complicated upper limb arteriopathies, 25% had a recognized occupational disease. Since December 1992, Erasmus' syndrome and systemic scleroderma due to silica without silicosis have been accepted in France as being occupational diseases. The therapeutic problems are discussed, notably the surgical indications in hypothenar hammer syndrome, the possible medical treatments of finger necrosis occurring in Erasmus' syndrome and, in particular, the role played by spinal cord neurostimulation. PMID- 8235150 TI - [Cancer of the anal canal; role of radiotherapy and combinations of chemotherapy and radiotherapy]. AB - Carcinoma of the anal canal is a rare disease, more common in women than in men. The mean age distribution at presentation is about 60 years. The natural history is mainly loco-regional, distant metastases being uncommon. Histologically, most cancer are of the squamous cell type of different keratinization. The pre treatment evaluation is mainly performed by methodical clinical examination. There is no widely accepted staging system for these tumours. Three statistically significant prognostic factors are admitted: tumour size, regional nodal involvement and histological grade. The french school (Tenon, Institut Gustave Roussy, Institut Curie, Lyon) and the experience at the "Princess Margaret Hospital" at Toronto have shown that irradiation of these tumours is an adequate therapy. In Europe, irradiation has always played a more important role in the therapy of these tumours, than in North America where surgery was often preferred as the initial therapy. With the introduction of combined modality treatment, the use of pre-operative concomitant radiochemotherapy, in North America, has again changed the treatment policy towards a conservative radiotherapeutic approach. The present study analyses the modalities and the results from radiation therapy alone and the preliminary results from concomitant irradiation and chemotherapy. PMID- 8235151 TI - [Para-septal emphysema and pseudo-interstitial syndrome]. PMID- 8235152 TI - [Incidental detection of retro-pneumoperitoneum in Cushing disease]. PMID- 8235153 TI - [Selective IgA deficiency and antiphospholipid syndrome]. PMID- 8235154 TI - [Acute pleuropericarditis with eosinophilia after ingestion of tryptophan]. PMID- 8235155 TI - [Arthropathy caused by fluoroquinolones in a patient after liver transplantation]. PMID- 8235156 TI - [Hepatocellular insufficiency and pseudo-surgical abdominal pain in adult onset Still disease]. PMID- 8235157 TI - [Emotional factors, growth, metabolism, and immunity]. PMID- 8235158 TI - [3 beta-hydroxysteroid dehydrogenase defect: frequency of presentation in a sample of Chilean hirsute women]. AB - The defect of 3 beta hydroxysteroid dehydrogenase (3 beta HSD) is frequent among hirsute women and clearly dependent on the ethnic composition of the studied population. Our aim was to study the frequency of 3 beta HSD deficit in a group of Chilean hirsute women. Basal and post ACTH concentrations of cortisol, 17 hydroxyprogesterone and 17 hydroxypregnenolone were measured by RIA in 40 hirsute post puberal women and in 15 normal age matched female volunteers. Criteria for considering a 3 beta HDS deficit were 17 hydroxypregnenolone values and 17 hydroxypregnenolone/17 hydroxyprogesterone and 17 hydroxypregnenolone/cortisol ratios after ACTH stimulation over the 95% confidence intervals of normal women. Basal dehydroepiandrosterone sulphate and testosterone levels were also measured in hirsute women. All samples were obtained during the follicular phase of the menstrual cycle. ACTH stimulated hormone values and ratios were diagnostic for 3 beta HDS deficit in 7.5% of hirsute women. Basal testosterone was over 80 ng/dl in 47.5% and dehydroepiandrosterone sulphate over 3.9 micrograms/ml in 52.5% of these women. There was no correlation between dehydroepiandrosterone or testosterone values and ACTH stimulated hormone values. It is concluded that 3 beta HSD is frequent in hirsute women and that its diagnosis requires the determination of ACTH stimulated 17 hydroxypregnenolone values and 17 hydroxypregnenolone/17 hydroxyprogesterone ratio. PMID- 8235159 TI - [Hemolysis induced by Loxosceles laeta venom. In vitro experience]. AB - To study the effects of loxosceles laeta venom on red blood cells and the possible factors involved in hemolysis during arachnidism, in vitro models were designed to measure the role of loxosceles venom, calcium, complement and antibodies in the mechanism of red blood cell destruction. The degree of basal hemolysis was measured in a 5% suspension of group O, Rh (+) red blood cells in pH 7.4 buffer. In a similar suspension spider venom was added in amounts equivalent to one venom gland. After 72 hours of incubation, basal hemolysis was 5.59 +/- 2.04% which increased to 26.01 +/- 7.9% adding venom (p < 0.001). Adding calcium to the incubation medium, hemolysis increased to 88.5 +/- 7.16% (p < 0.001). Incubating red blood cells with control human serum and venom, hemolysis was 14.58 +/- 2.42%, which decreased significantly to 6.85 +/- 3.35% when serum was heat inactivated; this demonstrates an effect of the presence of complement. We did not find antivenom antibody production in patients with arachnidism 10, 15 or 30 days after the spider bite. It is thus demonstrated that loxosceles laeta venom has a direct lytic action on red blood cells that is calcium and complement dependent and is not mediated by antibodies or other substance. PMID- 8235160 TI - [Epithelial lesions associated with gallbladder carcinoma. A methodical study of 32 cases]. AB - Using mapping techniques, 32 consecutive gallbladder carcinomas diagnosed in our department were studied. The defined lesions were dysplasia, carcinoma in situ and superficial spreading carcinoma. A total of 2257 lesions were recorded from 1072 inclusions. All the defined lesions were present in about 10% of all sections. However, if each case is considered separately, dysplasia was present in 81.3% of cases, carcinoma in situ in 65.6% of cases and superficial spreading carcinoma in 68.8% of cases. An isolated focus of atypical epithelium without anatomic relation with the main tumor was demonstrated in only 3 cases and a close relation between invasive carcinoma and dysplastic lesions was observed in the remainder. Infiltration of Rokistansky-Aschoff sinuses was observed in 40% of cases. We were not able to demonstrate, on histological basis, differences between superficial spreading carcinoma and carcinoma in situ. We conclude that atypical lesions of the gallbladder are not frequently seen and that their presence must prompt the search of a carcinomatous lesion by means of serial sections of all the histological sample. PMID- 8235161 TI - [Is it necessary to supplement with folic acid patients in chronic dialysis treated with erythropoietin?]. AB - The need for folate supplementation in patients on chronic hemodialysis receiving erythropoietin (EPO) remains to be determined. Thirty five patients on chronic hemodialysis were studied; of these 10 did not receive EPO nor folic acid, 12 received EPO with folic acid supplementation and the rest only EPO. In these groups, after 9 +/- 2.9 months of treatment, serum folate levels were normal, although higher in those patients supplemented with folate. An additional group of 8 patients, previously supplemented with 2 mg/week of folate, was studied during the first 10 weeks of EPO treatment. In these patients a significant decrease in serum folate was observed from the first to the tenth week (from 18 +/- 29 to 7 +/- 4 ng/ml). Red cell folate had an unexplained raise during the first four weeks and went back to near basal levels during the next weeks. As expected serum ferritin levels decreased at the end of the study period, but remained over 100 ng/ml Red blood cell protoporphyrin remained normal. We thus recommend the measurement of serum and red cell folate levels during the first and tenth weeks of the induction phase of EPO treatment. Also, folic acid supplementation in doses of 2 mg/week is recommended to maintain adequate body stores, specially in extremely anorectic hemodialysis patients or those in whom strict diets without fruits are prescribed. PMID- 8235162 TI - [Prospective study on post-acute myocardial infarction patients]. AB - Long-term prognosis in acute myocardial infarction (AMI) can be related to a number of risk factors, including the electrocardiographic stress test (ECG Test). This study analyzes its contribution to the prognosis and management of such patients. 60 individuals discharged from a coronary care unit, without or with persisting complications underwent an ECG test 4-6 weeks following AMI, and were followed for a period of 12-39 months. 40 cases had a negative ECG test. During follow-up they continued to be generally asymptomatic and their survival was 100%. Coronary angiography in patients younger than 45 years revealed mainly single vessel disease. 20 cases (33%) had a positive ECG test. During follow-up 50% of these had anginal manifestations, and coronary angiograms revealed mainly 2-3 vessel disease. 6 patients underwent surgical revascularization. One patient died awaiting surgery. Overall survival in this group was 95%. It is concluded that stress testing performed during the convalescence period of patients with an AMI allows recognition of patients with a potentially poorer prognosis, which can be modified by appropriate study and revascularization in selected cases, bringing survival expectations to the same level of more benign cases. PMID- 8235163 TI - [Diagnosis of bacterial meningitis by latex agglutination tests]. AB - Fifty samples of cerebrospinal fluid (CSF) from patients with acute bacterial meningitis were studied with direct gram examination, culture and latex agglutination. Twenty one of these samples had previous antimicrobial treatment. In 42 patients, blood cultures were also made. Thirty control CSF samples were also studied. The diagnostic yield of CSF culture was 72%, of direct gram examination 74% and latex agglutination 80%. All the techniques as a whole has a diagnostic yield of 92%. In the 21 CSF samples with previous antimicrobial treatment, culture was positive in 47.6% and latex agglutination in 85.7%. All control CSF samples were negative. It is concluded that the diagnostic yield of latex agglutination test in patients with previous antimicrobial treatment is significantly better than culture. PMID- 8235164 TI - [Results of surgical treatment and prognosis of colonic neoplasms complicated by perforation]. AB - Twenty two patients (13 males), whose ages ranged between 25 and 94 years, were subjected to surgical treatment for perforative carcinoma of the colon in a period of 10 years. Seventy three percent of tumors were localized in rectum and sigmoid colon. The perforation lied within the tumor in half of the patients and proximal to the tumor, specially in the cecum, in the rest. The perforation produced a local abscess in 5 patients and a diffuse peritonitis in 17 patients. Fifteen patients were subjected to segmental resections with a 40% operative mortality rate and 5 to a fecal diversion procedure with a 57% operative mortality. Tumor staging was similar to that of patients subjected to elective surgery. Crude 5 year survival was 18% and, excluding operative mortality, 57%. PMID- 8235165 TI - [Fatigue resistance of inspiratory muscles in patients with severe mitral valve stenosis]. AB - The strength and endurance of the inspiratory muscles was assessed in 14 patients with severe mitral valve stenosis uncomplicated by other diseases. Strength was evaluated measuring the maximal static inspiratory pressure (MIP). Endurance was measured using a two minute weight incremental test, to obtain the maximal sustainable pressure (SIP), which is the highest pressure that a subject can generate to mobilize air with incremental weight during inspiration. MIP was similar to that of 8 normal subjects (110 +/- 21 and 128 +/- 27 cm H2O respectively p = NS). SIP, maximal sustainable weight and SIP as percentage of MIP were 47 +/- 10 cm H2O, 261 +/- 81 g and 43 +/- 6% respectively, all lower than in normal subjects (89 +/- 25 cm H2O, 525 +/- 167 g and 71 +/- 15% respectively) We conclude that patients with severe mitral valve stenosis have decreased inspiratory muscle endurance compared to normal subjects and this feature may be related to their decreased exercise tolerance. PMID- 8235166 TI - [Mortality in an intensive care unit: predictive value of APACHE II severity score versus maximum APACHE]. AB - Patients admitted to intensive care units (ICU) experience constant changes in their general condition. Therefore, the determination of Apache score within the first 24 hours of admission may not be a reliable index of severity. The aim of this study was to measure daily Apache scores in ICU patients, and to determine if the maximum score (Maximum Apache) attained during hospitalization had a better prognostic value than that of admission. We measured daily Apache scores to 314 patients admitted to Hospital Clinico UC ICU. Mean admission score was 17.5 +/- 9.3 (range: 1-47) and mean maximum score was 18.6 +/- 9.7 (range: 1-47), p < 0.001. Mortality during ICU stay was 17.2%, and 8.6% during hospitalization after ICU discharge. Only 78.3% of patients attained their maximum apache score during the first 24 hours of admission, whereas 21.7% attained it during the rest of ICU hospitalization. Excluding subjects with less than one day of ICU stay, 33% of patients attained maximum Apache score after 24 hours of ICU admission. Statistical analysis showed that maximum Apache score was a better predictor of mortality than that of admission. PMID- 8235167 TI - [Nail-patella syndrome associated with monoclonal gammopathy. Review regarding a case]. AB - A 54 years old female with significant impairment of renal function is presented. On physical examination, the presence of dystrophic nails, elbow dysplasia and prominent iliac horns. Familial study showed similar nail and bone deformities in 3 of six sons. The nail patella syndrome diagnosis was based on these findings. This is a rare autosomal dominant hereditary disease, probably related to congenital alterations in collagen metabolism. Clinical characteristics include bone abnormalities that principally involve knees and elbows, nail alterations and the presence of iliac horns, that are considered pathognomonic of the syndrome. Renal involvement is observed in 30 to 55% of cases. This patient had also an IgA-lambda paraprotein, whose relationship to the above mentioned syndrome is uncertain, since no evidences of malignant plasma cell dyscrasia were demonstrated. PMID- 8235168 TI - [Disseminated histoplasmosis in a patient with acquired immunodeficiency syndrome]. AB - A twenty nine year old male homosexual presented with malaise, weight loss, fever and profuse sweating. An ill defined abdominal mass was found during physical examination in the right lower quadrant and chest X rays disclosed a pleural effusion. HIV antibodies and hepatitis B surface antigen were positive and immunological parameters were altered. Light and electron microscopic examination of operative biopsies of the abdominal mass revealed the presence of Histoplasma capsulatum. Treatment with Amphotericin B was started with a favorable response and the patient was discharged. He was readmitted with a septic shock and died. Necropsy showed pulmonary histoplasmosis. This is the first case of disseminated histoplasmosis in a patient with AIDS described in Chile. PMID- 8235169 TI - [Colonic neoplasms complicated by perforation]. PMID- 8235170 TI - [Significance of Helicobacter pylori infection in the pathogenesis and natural history of peptic ulcer]. AB - Recent studies strongly suggest that Helicobacter pylori might have a significant role in the pathogenesis and rate of recurrence of peptic duodenal ulcer. Various factors, such as the existence of ulcerogenic strains of the bacteria and susceptible subjects, might explain the fact that not more than 10% of the patients infected with H pylori actually have duodenal ulcer. Its association with gastric ulcer and gastric carcinoma is not well defined. PMID- 8235171 TI - [Neurobiological bases of schizophrenia]. AB - Environmental and biological factors play an important role in the etiology of Schizophrenia. We believe that the disorder is probably multifactorial with a genetic predisposition that triggers morphological (CT, MRN) and functional (EP, SPECT and PET) brain alterations in the areas, where the integration of the affect, comprehension and expression of emotions occur. These areas are also involved in the planification and organization of cognitive and emotional functions. These findings may have clinical implicancies (soft signs), and neurochemical corroborations (neurotransmitters and receptors qualifications and imaging in SPECT and PET. PMID- 8235172 TI - [Vasodilator agents in chronic heart failure: which is the best option?]. AB - Vasodilator therapy has been utilized for the treatment of congestive heart failure in the last 20 years. These drugs contribute to increase cardiac output, decrease peripheral vascular resistance and favour venous dilatation. Recent multicenter trials have addressed the issue of the impact of vasodilator therapy upon survival. Thus, the VHEFT-I and Consensus studies have shown that both the combination of nitrates and hydralazine and ACF inhibitors improve life expectancy in patients with moderate and severe heart failure. Moreover, the SOLVD study showed that ACE inhibitors improve survival and reduce cardiac events in patients with mild heart failure and depressed myocardial function at the end of 2 years of follow-up. The VHEFT II trial compared the effects of the nitrate hydralazine combination versus ACE inhibitors upon the clinical course of patients with moderate heart failure. This last trial showed that although nitrates and hydralazine exerted a slightly better benefit upon exercise tolerance and left ventricular ejection fraction, patients that were treated with ACE inhibitors had a significantly reduced mortality. Differences in mortality when both groups of vasodilators drugs were compared were due to reduction of arrhythmias and sudden death. It is likely that this greater benefit obtained with ACE inhibitors when compared to nitrates and hydralazine in heart failure might be due to their favourable effects upon the abnormal neurohormonal activation observed in this syndrome. Thus ACE inhibitors have turned out to be one of the cornerstones in the treatment of congestive heart failure. PMID- 8235173 TI - [Role and responsibility of the professions in the medical act and health actions]. AB - The medical act is a part of health actions, but not every health action is a medical act. The primarily and most generalized medical act is the care of a sick individual. The requirements to execute a medical act are to possess the specific knowledges and skills, have a recognition of the possession of these qualities and to be qualified by the state to practice the profession. The governing condition of medical task is the disease and the essential part is diagnosis and treatment. The diagnosis and treatment prescription are exclusively medical acts. The execution of treatment may be shared with other health care professionals. The actions of health promotion, prevention, repair and rehabilitation must necessarily be performed by a multi professional team in which the physician (or the odontologist) has predominantly the task of repairing the lost health. The diversification of health professions has created interprofessional tensions, with an underlying social power problem. Nevertheless, each profession has its own dignity and specific action boundaries. Although there are always unavoidable superpositions in their respective frontier limits role. PMID- 8235174 TI - [Poisoning by the consumption of shellfish contaminated with paralytic venom in the XII Region, Chile. Anatomopathological study]. AB - The necropsy findings of ten subjects died as a consequence of paralytic shellfish intoxication are presented. These deaths occurred between march 1991 and january 1992, affected to seamen and occurred within 72 hours of contaminated shellfish ingestion. Necropsies were performed within 24 hours of death. The most outstanding necropsy findings were the presence of pink lividities, mydriasis and isocoria, airway obstruction with gastric contents, severe brain, lung liver and spleen swelling and edema, and digestive and respiratory mucosal congestion and friability. The toxicological study of urine, gastric content and organ samples revealed the presence of shellfish paralytic poison. PMID- 8235175 TI - [Alcohol drinking in parents and its relation with intellectual score of their children]. AB - Among children living in orphanages of the Metropolitan Region, the paternal alcohol ingestion patterns were investigated. According to abnormal parental alcohol ingestion, a sample of 291 children was studied, of whom 32% were living in simple protection homes for infants with normal intellectual coefficient and 52% in homes for mentally retarded boys. There was a higher frequency and intensity of abnormal alcohol ingestion among parents of infants living in homes for the mentally retarded. Among these, 29% of mothers were illiterate and 11% had incomplete primary school education compared to mothers of simple protection homes in whom the frequency of illiteracy was 10% and of incomplete primary education, 50%. Twenty percent of fathers of children living in homes for the mentally retarded were unemployed, 16% had occasional jobs and 11% had a stable work; these numbers were 11, 30 and 15% respectively for fathers of simple protection homes. Among infants living in homes for the mentally retarded, undernutrition was found in 44%, growth retardation in 51% and microcephaly in 34% compared to 33, 30 and 11% in children living in simple protection homes. Sixteen percent of the sample had fetal alcohol syndrome, 24% among those living in homes for the mentally retarded and 12% in simple protection homes. There was an inverse relationship between offspring intellectual coefficient and the number of drinking parents. PMID- 8235176 TI - Verifying keyed medical research data. AB - Although diminished use of double keying for the verification of keyed medical research data has occurred, there are no published data that demonstrate the existence of equivalent or better replacement. The simple replacement of double keying with visual comparison is not recommended because visual comparison is too susceptible to the transient energy levels of the individual who makes the comparison. Some system design innovations applicable in specific situations allow elimination of verification by providing nearly equivalent or possibly even superior data quality assurance opportunities. These alternatives include designs where one replaces the transcription to data forms by direct keying from original source documents coupled with extensive quality reviews, and designs with direct data base input of data subjected to intense automated consistency checking and immediate analytic use. In both cases, the absence of keyed data verification places a greater burden for data quality assurance on other subsystems, such as quality review, and ultimately the analytic process. Data entry systems based on new technologies, such as tablet computers, electronic image processing, and voice input, will require similar considerations for assuring data quality. PMID- 8235177 TI - A regression model for the rate of success of in vitro fertilization. AB - The treatment of infertility by in vitro fertilization involves several aspects of human reproductive physiology. Since failures are still numerous and since improved techniques are becoming increasingly available, it is important to develop a model where the components of success are identified, allowing estimation of the occurrence of these components of success in the infertile population. The model of Speirs reduces these components to two: uterine receptivity and embryo viability. The probabilities of uterine receptivity and embryo viability were estimated here in a retrospective study of patients in a Belgian hospital sample. For comparison, Australian and U.S. data were used. It was shown that the estimate of uterine receptivity for the Belgian sample is located between the estimates for Australian and U.S. samples. In view of the psychological problem of repeated unsuccessful attempts at in vitro fertilization in some patients, it is important to be able to single out some prognostic factors that would allow more precise estimation of the probability for success in each case. A regression method was developed for that purpose. Only the effect of age, in the Belgian sample, was found to influence uterine receptivity significantly. PMID- 8235178 TI - A model for seasonal changes in time series regression relationships: with an application in psychiatry. AB - This paper proposes a two-stage linear model for the regression relationships between a time series and a set of time-dependent covariates when the relationships change according to a seasonal pattern. The first stage consists of a piecewise constant autoregression model for these relationships. The second stage models the piecewise coefficients with harmonic equations to examine for seasonal trends. This approach models the seasonal trends in time series regression relationships in the presence of non-constant autoregression, non constant residual variance, and stochastic variation in the regression effects across time. Seasonal affective disorder (SAD) time series data illustrate the methodology. I model the seasonally changing regression relationships between the change in daily sunlight (from the previous day) and depressive mood and between the change in daily temperature and depressive mood for a SAD patient, and I employ an empirical Bayes method to obtain efficient month-specific estimates for these relationships. PMID- 8235179 TI - Further results on the non-parametric linear regression model in survival analysis. AB - This paper gives further developments of a non-parametric linear regression model in survival analysis. Three subjects are studied. First, martingale residuals, originally developed for the Cox model, are introduced for our linear model. Their theory is developed and they are shown to be useful for judging goodness of fit. The second focus of the paper is on the use of bootstrap replications to judge which features of the cumulative regression plots are likely to reflect real phenomena and not merely random variation. In particular, this is applied to judging whether the effect of a covariate disappears over time, a problem for which no formal test exists. The third subject is density type, or kernel, estimation of the regression functions themselves. This might give more direct information than the cumulative plots. The approaches are illustrated by data from a clinical trial of carcinoma of the oropharynx, and by survival times of grafts in renal patients. PMID- 8235180 TI - Estimating the causal effect of smoking cessation in the presence of confounding factors using a rank preserving structural failure time model. AB - Estimating the causal effect of quitting smoking on time to death or first myocardial infarction requires that one control for the differences in risk factors between individuals who elect to quite at each time t versus those who elect to continue smoking at time t. In this paper we examine the limitations of standard time varying Cox proportional hazards models to yield tests and estimates of this effect. Implementing the method of G-estimation proposed by Robins, we perform an observational analysis of data from the Multiple Risk Factor Intervention Trial (MRFIT) and estimate the causal effect of cigarette cessation while controlling for such time varying confounders as angina. We reject the null hypothesis of no effect of quitting on time to failure, and estimate that by quitting smoking, an individual increases by 50 per cent his time to death or first myocardial infarction (MI). PMID- 8235181 TI - Incomplete factorial designs for randomized clinical trials. AB - Recently there has been increased interest in considering factorial designs for randomized clinical trials when one wishes to study two or more treatments. Such designs may offer impressive gains in efficiency compared with a series of trials studying one treatment at a time. This is especially true when the treatments do not interact with one another. If interactions are of special interest, factorial designs provide one sensible approach for studying them, but larger sample sizes would be required because tests for interactions have lower power than those for main effects. In trials designed to test putative agents for preventing cancer, interactions may be of less interest so that fractions of higher-order factorial designs might be appropriate. Sometimes it may not be reasonable, interesting, feasible, or ethical to study all treatment combinations required in a complete or fractional factorial design, yet one may want to preserve some of the factorial structure to increase efficiency and to aid understanding. For such situations, incomplete factorial designs are proposed. Although not all of the advantages of full factorial designs are preserved, such designs may provide reasonable compromises for certain situations. PMID- 8235182 TI - Approaches to sample size estimation in the design of clinical trials--a review. By A. Donner, Statistics in Medicine, 3, 199-214 (1984) PMID- 8235183 TI - [In memoriam Pierre Dustin]. PMID- 8235184 TI - [Mesenteric venous infarction and antithrombin III deficiency: description of 2 cases and literature review]. AB - Congenital deficiency of antithrombin III, the main physiological inhibitor of blood coagulation, enhances recurrent venous thromboses, usually in the mesenteric network. Two such cases are reported. PMID- 8235185 TI - [Monitoring of fontanelle pressure in neonates and infants. Evaluation of a new measuring technique, determination of normal values and clinical usefulness]. AB - The experience with 200 measurements of anterior fontanelle pressure with the Rotterdam Teletransducer in newborns and infants is reported. Statistical analysis of 25 comparative measurements between anterior fontanelle pressure and invasive cerebrospinal fluid pressure showed an excellent correlation (rs = 0.95). Measurements were reproducible and the plot quality allowed visualisation of pulse pressure and pressure waves. Normal values of the anterior fontanelle pressure, pulse pressure amplitude and pressure wave maximal amplitude were established in 15 prematures, 27 term newborns and 10 infants. Anterior fontanelle pressure monitoring was performed in 19 term newborns with post asphyxial encephalopathy, 18 newborns and infants with hydrocephalus, 8 preterm and term newborns with respiratory distress and 19 patients with subdural haematomas, metabolic diseases, meningitis, subarachnoidal haemorrhage, head trauma, post cardiac arrest encephalopathy and abnormal head growth or bulging fontanelle. Abnormal patterns of anterior fontanelle pressure monitoring were found in moderate or severe neonatal post-asphyxial encephalopathy, evolutive hydrocephalus, subdural haematomas, metabolic diseases with hyperammoniemia and other clinical situations. In contrast, anterior fontanelle pressure monitoring yielded normal values in mild post-asphyxial encephalopathy, arrested hydrocephalus, well functioning ventriculo-peritoneal derivation, and in normal infants with rapid head growth or bulging fontanelle. The Rotterdam Teletransducer provides thus accurate and reproducible values of intracranial pressure. Anterior fontanelle pressure monitoring may be of value in many situations in clinical practice. PMID- 8235186 TI - [Ignorance and misconceptions in the use of contraceptive methods]. AB - Good knowledge of the available contraceptive methods ensures better acceptability, continuity and satisfaction, and lower failure rates of these methods. This paper describes an inquiry on the contraceptive knowledge in 114 women attending three Brussels family planning clinics. Misconceptions, beliefs in false rumors, and considerable insufficiencies in knowledge were detected. In order to increase patient's ability to correctly use birth control methods, so as to lower unwanted pregnancy rate, efforts should be made by health practitioners as well as by mass media and teaching institutions. PMID- 8235187 TI - [Superior mediastinal abnormality in thoracic radiography]. PMID- 8235188 TI - [Medical secret and insurance]. PMID- 8235189 TI - New frontiers in thermoregulation and exercise. PMID- 8235190 TI - Biomechanics of rugby union scrummaging. Technical and safety issues. AB - In the game of ruby union, the scrum epitomises the physical nature of the game. It is both a powerful offensive skill, affording a base for attacking play, and a defensive skill in denying the opposition clean possession. However, the scrum has also been implicated in a large proportion of serious spinal injuries in rugby union. The majority of injuries are found to occur at engagement where the forces experienced by front-row players (more than two-thirds of a tonne shared across the front-row) can exceed the structural limits of the cervical spine. These large forces are a consequence of the speed of engagement and the weight (and number) of players involved in the scrum. This highlights not only the need for physical preparation of all forwards but particularly player restraint at engagement, and justifies the 'crouch-pause-engage' sequence recently introduced to 'depower' the scrum. As the hooker is the player exposed to the greatest loads throughout the scrum and subsequently most at risk, he should determine the timing of engagement of the 2 front-rows. Stability of the scrum is an indication of front-row players' ability to utilise their strength to transmit the force to their opponents as well as the push of second-row and back-row players behind them in the scrum. This appears to be independent of the size of players. Equally, it reflects the risk of chronic degeneration of the musculoskeletal system through repeated exposure to these large stresses. However, not only are older and more experienced players better able to generate and transmit these forces, they are also able to maintain the integrity of the scrum. A large proportion of individual players' efforts to generate force is lost in their coordinated effort in a normal scrum. It is assumed these forces are dissipated through players re-orientating their bodies in the scrum situation as well as to less efficient shear forces and to the elastic and compressive tissues in the body. It again reinforces the importance of physical preparation for all forwards to better withstand the large forces involved in scrummaging. Despite negative publicity surrounding the risk of serious spinal injury in rugby union, limited research has been conducted to examine either the mechanisms of injury or techniques implicated in causing injury. Biomechanical information can provide systematic bases for modifying existing techniques and assessing the physical capacities necessary to efficiently and safely play in the serum. This will both improve performance of game skills and minimise the potential for injury. PMID- 8235191 TI - Applied physiology of female soccer players. AB - Women's soccer is in its infancy as a sport and research into its physiological demands and the physical characteristics of players is somewhat limited. There is now an increasing demand for scientific investigation of the female game and of the players, match analysis and role variations requiring particular attention. Current research suggests that the demands of the game for women are similar to those placed on male players. Women are reported to cover a similar distance (mean 8471m) to their male counterparts during a game and much the same proportions of the game appear to be devoted to exercise of varying intensities. Furthermore, female and male players appear to tax the aerobic and anaerobic energy systems to a similar level. The physical and physiological characteristics of female soccer players are comparable with those of other female games players and are more favourable than average for the population. Mean body fat percentages of between 19.7 and 22.0% and VO2max values of between 47.1 and 57.6 ml/kg/min have been reported for elite female players, while faster than average sprint times are also characteristic of them. PMID- 8235192 TI - Amino acid supplementation and exercise performance. Analysis of the proposed ergogenic value. AB - Increasing the relative amount of protein in the diet of athletes has been suggested to optimise anabolic processes and improve both physiological responses to training and performance. While energy balance studies generally support the concept that athletes may require additional protein in their diets in comparison with the Recommended Dietary Allowances (RDA), most sport nutritionists contend that as long as athletes maintain energy balance and ingest 15% of their total caloric intake in the form of protein, additional supplementation of protein is not necessary. Recently, amino acids have become a popular nutritional supplement marketed to athletes. In strength athletes, amino acid supplementation has been proposed to increase the availability of essential amino acids, enhance anabolic processes promoting tissue accretion, and accelerate the rate of recovery during training. In endurance athletes, amino acid supplementation has been proposed to improve physiological and psychological responses during endurance exercise and training. There appears to be little scientific evidence to support the hypothesis that amino acid supplementation may enhance the physiological responses to strength training when athletes consume dietary protein within the recommended guidelines. Results of the effects of amino acid supplementation on the physiological and psychological responses to endurance exercise are preliminary. However, the findings suggest that amino acid supplementation with carbohydrate before, during and after exercise may alter the ratio of free tryptophan to branch-chained amino acids. Further research is required before definitive conclusions can be drawn regarding the proposed ergogenic value of amino acid supplementation. PMID- 8235193 TI - The effect of taping on ankle stability. Practical implications. AB - Ankle ligamentous injuries are very common in sports. One of the most popular methods of supporting the weakened ankle is external support with ankle tape. The tape can be used to prevent ankle injuries, to stabilise the injured ankle to prevent further injury, and as a stabiliser during the rehabilitation phase. Of the various taping methods, basket weaves (combined with stirrup and a heel-lock) are the most effective for increasing the mechanical stability of the ankle, especially in the frontal plane (talar tilt). A combination of increased mechanical stability, decreased range of ankle motion and an improvement in the proprioceptive function of the ankle ligaments are factors associated with effectiveness of ankle taping. The major drawbacks of ankle taping are the high costs and decreased support after exercise. PMID- 8235194 TI - Achilles tendon lesions in sport. AB - Achillodynia (Achilles tendon pain) is a significant source of disability to many people taking part in sports. Papers in the English language published since 1986 are reviewed here, grouped into specific subject areas including biomechanics, pathology, general clinical presentations, experimental treatments, steroids, podiatry and surgery. While there has been no dramatic breakthrough in the field, there have been various interesting advances with particular reference to imaging and conservative management, which will hopefully stimulate further studies. Many problems of Achilles tendon lesions in athletes remain unsolved, however, and much is yet to be done to provide adequate and generally effective methods of prevention and conservative treatment. PMID- 8235195 TI - The paradox of Parse's theory. PMID- 8235196 TI - The nurse-patient relationship. AB - My article considers elements of the philosophy of Buber and Marcel in relation to recent changes in nurse education which have potential for limiting the clinical contact time for student nurses. Under three headings, the nurse-patient relationship is discussed: time limitations, I-thou and I-It forms of relating, and the nature of presence. The literature supports the view that a nurse's openness or permeability in relating to patients in their real needs, is of major importance in meeting the patient's needs. The concept of presence indicates that changes occur in both parties in relationships in the influx of each other's presence. Being closed to the other's influx renders the relationship one of I-It in which objectification of the patient can occur--he or she is seen as a 'case'. Even counselling skills could fall into this category in a 'double glance'. PMID- 8235197 TI - The role conflict of the forensic nurse. PMID- 8235198 TI - Absent nurses. PMID- 8235199 TI - Care for the mentally ill. PMID- 8235200 TI - Nursing theory vs nursing practice. PMID- 8235201 TI - The nurse teacher's new clinical role. PMID- 8235202 TI - Comparing degree and diploma courses. PMID- 8235204 TI - A case for humanism in nurse education. PMID- 8235203 TI - The beginning of the end of andragogy. PMID- 8235205 TI - Nursing futures. PMID- 8235206 TI - Quality strategy in nurse education. PMID- 8235207 TI - Mental health in action. PMID- 8235208 TI - [Effects of the stimulation of the subthalamic nucleus in Parkinson disease]. AB - In Parkinson's disease, experimental studies favour a neuronal hyperactivity of the subthalamic nucleus. We carried out a subthalamic nucleus electrical stimulation in a patient aged 51, suffering for 8 years from a severe akineto rigid form of Parkinson's disease, complicated with an on-off effect. Stereotaxic surgery was done under local anaesthesia on one side. Within the theoretical target, a 130 Hz stimulation induced akinesia alleviation mainly on the contralateral limbs. No abnormal movement was noticed. Then a long-term quadripolar DBS Medtronic electrode was inserted in that area. The study of the effects of chronic stimulation is in progress to determine the best temporal and electrical stimulation variables. PMID- 8235209 TI - [Transcutaneous mechanical vibrations: analgesic effect and antinociceptive mechanisms]. AB - Although vibration has been used empirically for a long time in medicine, it was only about ten years ago that it began to be investigated scientifically. In the first part of this paper, we review the main data showing that vibratory stimulations can relieve both acute and chronic pain. In the second part, we consider the mechanisms responsible for the observed analgesic effects and discuss the nature of the sensory receptors involved and the effects of their activation on the triggering of central pain control systems. PMID- 8235210 TI - [A clinicopathological case of progressive aphasia]. AB - We report the case of a 59-year old right-handed woman presenting with progressive aphasia without any other neurological deficit and characterized by anomia, agramatism and auditory comprehension difficulties. CT scan showed no abnormalities. Four years later, aphasia was complete but neurological examination was normal. CT scan disclosed a mild cortico-subcortical cerebral atrophy with slight widening of both sylvian fissures. The results obtained with the Wisconsin Card Sorting Test and the Raven's Matrices showed only minor deficits. The IQ (WAIS) was 90. During the following 6 years the patient remained fully self-sufficient and carried out her homework normally. At that time MRI showed progression of cerebral atrophy more pronounced on the left side. Besides a rapid deterioration, twelve months later she developed severe dementia and died 13 years after the onset of the illness. Brain examination disclosed a severe atrophy (brain weight: 880 g) prominent in both frontal lobes and in the anterior perisylvian structures, more pronounced on the left side. There were no vascular lesions. Microscopy revealed widespread neuronal loss and astrocytic fibrillary gliosis confined to cortical areas and vacuolation in the superficial layers. Neurofibrillary tangles and neuritic plaques were found in the most atrophic areas but not in sufficient number to fulfill the histological criteria for Alzheimer's disease. There were no neuropathological changes of Pick's disease or subcortical degeneration. Previous microscopical studies of primary progressive aphasia showed non specific, mostly lobar atrophy similar to that observed in our case, although Alzheimer, Pick and Creutzfeldt-Jakob diseases have been reported. This neuropathological heterogeneity confirms that progressive aphasia is a non specific language disorder mostly observed in lobar forms of brain degeneration. PMID- 8235211 TI - [Polysaccharide bodies: an unusual finding in a case of temporal epilepsy. Review of the literature]. AB - Massive occurrence of polyglucosan bodies (PB) was found in the surgically removed temporal lobe of a 34 year-old woman presenting with complex partial seizures. The term of PB was proposed in order to group Lafora bodies (LB), corpora amylacea (CA) and Bielschowsky bodies (BB) on the basis of their biochemical similitude. A rigorous histochemical differentiation between these anomalies appears to be impossible at present. LB, BB and CA are mainly made up of glucose polymers i.e. polyglocusans. The pathway(s) leading from glycogen accumulation to PB formation is still unknown. PB are a hallmark of two diseases: Lafora disease and adult polyglucosan body disease. PB have occasionally been reported in rare cases of a variety of other neurological diseases. In all cases they were located within the neurons. This site characterizes Lafora bodies. BB are intraneuronal inclusions but restricted to neurons of the external pallidum. CA occur predominantly in the astroglia during the course of ageing. The significance of these structures depends on their regional distribution. The resemblance does not imply a common etiology for all conditions in which such bodies occur; it is probably due to the sharing of the final path in their causative pathway. Our case does not correspond to any of the classical diseases in which PB have been found. In our opinion, the patient exhibited a localized form of glycogen storage disease. PMID- 8235212 TI - [Tick-borne encephalitis in Alsace]. AB - Central European tick-borne encephalitis is mainly found in Central European countries and Austria where hundreds of cases are reported each year. Apart from 2 cases diagnosed in Alsace in 1968 and 1970 respectively, this disease was hitherto unknown in France. We report 8 new cases observed in Alsace between 1985 and 1990. Clinical presentation in these 10 patients was a pure meningitis syndrome in 4 cases and meningo-encephalitis in 6 cases, very severe in 3 of them. All patients recovered rapidly, and only 3 have slight sequelae. In a seroprevalence survey conducted in 1989 among 619 professional foresters of Eastern France, 8% were found to be seropositive, which suggests that the disease is often unrecognized. A study of the large series published in Austria and in other Central European countries has shown that the prognosis of tick-borne encephalitis is not always as favourable as it was in the Alsatian cases: severe sequelae or death occur in 1 to 2% of the patients. The need for a better detection of the disease and for vaccination of the subjects at risk must be emphasized. PMID- 8235213 TI - [Hemifacial spasm treated with botulinum toxin]. AB - Botulinum toxin produces a transient presynaptic focal block at the neuromuscular junction. Thus it induces muscle weakness which has a significant beneficial effect on hemifacial spasm. Fifty-four patients were treated and received at least 2 and at the most 22 consecutive injections over a 5-year period. Injections were repeated every 9 weeks on average. The interval between injections corresponded to the time elapsed between the last injection and the re emergence of spasms. Nineteen patients abandoned the treatment for various reasons. The most frequent side-effect was a ptosis which was observed in 1 out of 6 injections. Ptosis is due to diffusion of the botulinum toxin over a territory larger than expected. The results obtained in 42 patients were analyzed: there was no effect in 7 patients (17%); 11 patients improved by less than 50%; 13 by 50 to 70% and 11 more than 75%. PMID- 8235214 TI - [Visual evoked potentials and face recognition. Influence of celebrity and emotional expression]. AB - Visual evoked potentials (VEP) were recorded in the right and left parietal and occipital regions of 40 right-handed controls in a facial recognition task. VEP were studied first according to the renown of the faces, then according to their emotional expression. Asymmetry was noted between the hemispheres: P100 was of greater amplitude and longer latency in the left occipital region. Later components (P400 and P600) were of greater amplitude and longer latency in the right parietal region in all situations. P100 latency on the left side was shorter for renowned faces than for non-renowned faces (P = 0.05). P600 latency was shorter on the right (P < 0.03) and left (P < 0.05) sides for smiling than for non-smiling faces. When the subjects were asked to look for emotional expression of the face (smiling or non-smiling) P400 was very ample and P600 of little amplitude. When the subjects were asked to recognize the face (renowned or not renowned) P600 was very ample and P400 of little amplitude. Thus, there seems to be a differential treatment of information: automatic and rapid to detect emotion (P400), controlled, tardy (P600) and involving memory in the search for renown. PMID- 8235215 TI - Wernicke-Korsakoff syndrome following attempted hanging. AB - The case of an alcoholic man is reported who survived a suicidal hanging attempt, and subsequently suffered of amnesia, dementia, apathy and behavioural abnormalities. A clinical diagnosis of hypoxic brain damage was made, but upon autopsy six years later, extensive pseudosystematic thalamic degeneration and mammillary body atrophy were found, indicating a status post Wernicke encephalopathy. Precipitation of the disease is attributed to the synergetic effect of cerebral hypoxia/ischemia and thiamine deficiency. PMID- 8235216 TI - [Iatrogenic axonal neuropathy and erythroderma induced by piroxicam. Manifestations of hypersensitivity?]. AB - A case of peripheral neuropathy following treatment with piroxicam is reported. The patient developed muscle pain and asymmetrical sensory distal neuropathy of the lower limbs a few months after the onset of the treatment. Piroxicam was withdrawn and full recovery was achieved within one year. The simultaneous appearance in the same patient of erythroderma with microvasculitis on nerve and skin biopsies suggests a common type III hypersensitivity mechanism in the pathogenesis of these two side-effects of piroxicam. PMID- 8235217 TI - [Familial complex myxoma-lentiginosis disclosed by probable transient ischemic accident]. AB - A probable transient ischemic attack in a 45-year-old woman revealed bilateral atrial myxoma associated with lentiginosis. The patient's 20-year-old son had isolated lentiginosis. The characteristics of this autosomal dominant syndrome are reviewed. PMID- 8235218 TI - [Primary lymphoma of the central nervous system. Multiple remission with corticosteroid therapy]. AB - A primary T-cell lymphoma of the central nervous system was diagnosed at autopsy in a 73-year-old woman. The course of the illness was made of multiple neurological episodes: hypersomnia, palsies of cranial nerves, aphasia, hemiparesis, epileptic fits during 4 years, with fast recoveries under corticosteroid therapy. CT scan and MRI abnormalities were also dramatically improved by the treatment. PMID- 8235219 TI - [Transient neurologic deficit preceding intracerebral hemorrhage. Physiopathological hypotheses]. AB - Seventeen out of 238 cases (7%) of intracerebral hemorrhage were preceded by transient neurological deficit. In two cases, intracerebral hemorrhage and transient neurological deficit were thought to be due to amyloid angiopathy, this diagnosis was confirmed at autopsy in one case. In one case, intracerebral hemorrhage and transient neurological deficit were due to a cavernoma. In the other 14 cases, hypertension was the major risk factor. In 3 of them, CT-scans made after the transient neurological deficit but before the intracerebral hemorrhage showed deep small hypodensities suggestive of lacunas. The mechanisms of transient neurological deficit preceding intracerebral hemorrhage are reviewed. PMID- 8235220 TI - [Arnold-Chiari malformation in adults. Development of diagnostic conditions since the availability of MRI]. AB - We reviewed retrospectively 47 patients with a Chiari malformation (CM) who had been admitted to the Clinique de Neurologie of Lyon from 1971 to 1991. The purpose of this report is to assess the possible influence on the frequency and the clinical pattern at presentation of CM since 1985 when Magnetic Resonance Imaging (MRI) became available to us. A diagnosis of CM was made for 21 patients before 1985 (group 1) and for 26 patients thereafter (group 2), i.e., a 2.5 increase of incidence of CM frequency (1.5 patients/year before 1985 vs 3.7 patients/year thereafter). This increase was even more striking for cases without syringomyelic symptoms and signs (SSS) (42.9% in group 1, vs 76.9% in group 2). The accuracy of clinical diagnosis was 68.1% for the total surveyed period (90.5% before 1985 and 50% thereafter). For cases with SS at presentation, a syringomyelic cavity was radiologically demonstrated in 80% of the cases in the pre-MRI period and 90.9% of the cases since MRI was available. On the other hand, when a syringomyelic cavity was radiologically demonstrated, there was a related clinical SS in 70% of group 1 patients and 50% of group 2 patients. PMID- 8235221 TI - [Adrenoleukomyeloneuropathy in an adult]. AB - We report a case of adrenomyeloneuropathy in a 35-year-old man. He experienced acute adrenal insufficiency at 29 years of age, followed by psychic disorders and mental deterioration, then spastic paraparesia, dysarthria and peripheral neuropathy. MRI showed high-intensity signal areas in the cerebellum and periventricular white matter. Clinical symptoms, neurophysiological and neuroradiological examinations disclosed both features of adrenoleukodystrophy and adrenomyeloneuropathy. This indicates that adrenoleukodystrophy and adrenomyeloneuropathy probably represent two portions of the same entity. PMID- 8235222 TI - [Agenesis of the corpus callosum and epilepsy. 26 cases]. AB - Agenesis of the corpus callosum is a cerebral malformation diagnosed by CT scan. Beside mental retardation, the most frequent clinical manifestation is epilepsy. In a personal study of 26 cases, the authors analysed the clinical features of epileptic seizures observed with this malformation. They put the stress on the paradoxical presence of generalized seizures. PMID- 8235223 TI - [Ataxic monoparesis of the upper limb and suspension of the tonus caused by parietal lesion]. AB - A 58-year old right-handed man with chronic non valvular atrial fibrillation developed an acute left upper limb ataxia with transient numbness and mild motor impairement. Two weeks later, there was a severe degree of ataxia of the left upper limb and prominent asterixis of the left hand. CT scan and MRI showed a small parietal infarct. Initial median nerve somatosensory evoked potentials (SEPs) showed mild impairement of right parietal responses with absent right frontal SEPs P22 and N30. Two months later, parietal responses were normal but right frontal SEPs P22 and N30 remained abolished while ataxia of left upper limb persisted. Electromyographic activity recorded at the same time showed periodic involvement of the left hand distal tonus. These finding suggested that both ataxia and asterixis were due to a single postcentral infarct. Frontal SEP components are known to convey proprioceptive inputs which could be received by neocerebellar afferent pathways. Generators of these components are presumably located in premotor cortex and can be activated through parietofrontal connections. In our case it can be assumed that the parietal infarct involved these connections, which are mainly implied in the regulation of postural tonus of the distal upper limbs, and simultaneously impaired neocerebellar afferent pathways resulting in the emergence of parietal ataxia. PMID- 8235224 TI - [Bulbar hematoma of the olivary area. An analysis of the autonomic syndrome]. AB - The consequences of an anterolateral medullary hematoma in a 50 year old patient are reported. The lesions principally involved the inferior olive nuclei, the lateral reticular nucleus, and, partially, the nucleus ambiguus. Long-lasting postural control deficit and hypotonia were observed, associated with alpha motoneurons hyperexcitability. Analysis of the heart rate regulation showed a severe reduction of the normal accelerating reactions and a relative preservation of slowing down reactions that suggested a deficit of the ortho-sympathetic control. Unilateral palsy of the tongue and pharynx was also observed without sensory or sensorial deficit. The long term course was favourable. PMID- 8235225 TI - [Emotional disorders in HIV infection]. AB - Psychiatric and neurological symptoms have been the target of many studies, but the emotional and behavioral modifications in HIV infection remain quite unknown. Working on the emotional dimensions and the heterogeneity of depressive mood, we were interested in evaluating the emotional symptomatology in HIV infected patients. Fifteen HIV-positive and fifteen HIV-negative homosexual men paired by age and educational level were studied. They were seen by two trained psychologists who assessed depression, anxiety and mood dimensions with the MADRS depression scale, Covi's anxiety scale, Depressive Mood scale, Abrams and Taylor scale for Emotional Blunting and Retardation scale. HIV-positive subjects had significantly higher scores of emotional blunting: anhedonia and hypoexpressiveness. Scores of depression, anxiety, irritability and hyperexpressiveness were not significantly different between both groups. Hypoexpressiveness scores were correlated to the CDC stages of the disease. This means that the emotional deficit seems to increase with the course of the disease, and is present in the absence of depression or anxiety. The question of the origin of this emotional blunting can be raised: is it the result of an adaptative behavior and/or the action of the virus on the central nervous system? Further studies are needed to confirm these results and answer this question. PMID- 8235226 TI - [Demyelinating nervous lesions in myalgia-eosinophilia syndrome induced by L Tryptophan]. AB - A case of severe peripheral nerve lesions in a patient with eosinophilia-myalgia syndrome induced by L-Tryptophan is reported. Both superficial radial nerves were very tender on palpation. Biopsy of the radial nerve showed myelin abnormalities of the type observed in experimental chronic nerve entrapment. Nerve lesions included asymmetrical distortion of the myelin sheath, segmental demyelination, axonal degeneration and regeneration. Such abnormalities suggest that mechanical factors, presumably due to proliferation of connective tissue, play a role in the nerve lesions observed in this patient. PMID- 8235227 TI - [4 new cases of neuromyositis, one of them associated with HTLV-I infection]. AB - Neuromyositis is a very rare type of polymyositis where, beside the usual muscular manifestations, there are signs of peripheral neuropathy which can be found at clinical, electromyographic and/or pathological examination. We have seen between 1983 and 1990 four cases of neuromyositis. The neurological disorder was an axonopathy in two cases which is usual in neuromyositis; in the other two cases, there was a polyradiculoneuritis which seems to be very rare in this syndrome. In the four patients the disease was particularly severe and unresponsive to treatment. One of our patients had HTLV-I infection diagnosed by polymerase chain reaction amplification and in situ hybridization, while the serological test was negative. To our knowledge only three cases of HTLV-I associated neuromyositis have been reported in the literature. PMID- 8235228 TI - [Cerebral infarction, cardiac myxoma and lentiginosis]. AB - Occlusion of the middle cerebral artery in a 16-year old woman revealed a left atrial myxoma associated with a mucocutaneous lentiginosis. Two-dimensional transthoracic or transesophageal echocardiography should be performed in all patients with stroke and lentiginosis to look for cardiac myxoma. After treatment of the cardiac tumor, echocardiography should be repeated at regular intervals to detect a recurrence of the myxoma. As this cardio-cutaneous syndrome is a familial disorder with autosomal dominant inheritance, examination of all family members by echocardiography is recommended. PMID- 8235229 TI - [Gelastic epilepsy. Probable diencephalic hamartoma]. AB - We report a of gelastic epilepsy. MRI showed a probable hamartoma in the diencephalic region. PMID- 8235230 TI - [Infarction of the red nucleus and crossed cerebellar diaschisis]. AB - We report a case of rubral infarct involving adjacent ipsilateral thalamic structures in a 54-year-old man with hypertension and coronary heart disease. Clinical picture at onset was suggestive of Weber's syndrome leading to a Claude's syndrome with vertical gaze palsy and sleep disturbances. Single photon emission computed tomography using Hm-PaO-Tc revealed a crossed cerebellar diaschisis. This is, to our knowledge, the first case of such crossed cerebellar diaschisis in a patient with rubral infarction. PMID- 8235231 TI - [Paraneoplastic cerebral venous thrombosis]. AB - A 45 year-old patient presented with an epidermoid carcinoma of the tongue and cranial hypertension. MRI showed dural venous sinus occlusion, without infiltration or compression by a metastatic tumor. Anticoagulation led to recanalization of the occluded sinuses, as shown by the follow-up MRI, three months later. The pathophysiology of these rare paraneoplastic cerebral venous thrombosis is discussed. PMID- 8235232 TI - [Unilateral infarction of the cervical spinal cord. 2 cases identified by MRI]. AB - We report two cases of unilateral upper cervical spinal cord infarction in the territory supplied by the anterior spinal artery. Early nonspecific MRI abnormalities consisted of hypointense T1 signal and hyperintense T2 signal. A late MRI study showed partial decrease in lesion size due to perifocal edema resorption with negative Gd-DTPA enhancement. The presumed cause of infarction was occlusion of a sulcocommissural artery or of the ipsilateral nutrient branch of the spinal artery. PMID- 8235233 TI - [Abnormalities of the muscular bioenergetics in Steinert's disease]. AB - The thenar muscles and gastrocnemius of a patient with myotonic dystrophy were investigated, at rest, by phosphorus nuclear magnetic resonance spectroscopy. A decrease in phosphocreatine level and an increase in inorganic phosphate and phosphodiester levels were found in the gastrocnemius, which was clinically spared, whilst the thenar muscles, which were wasted and affected by myotonia, exhibited only an increased inorganic phosphate level and an elevated pH. These findings were comparable with those found in other muscular disorders, such as Duchenne's and Becker's dystrophies, as well as in limb girdle dystrophy. They suggested that the abnormalities observed were unrelated to myotonia or wasting, and the possibility of a secondary mitochondrial disorder in myotonic dystrophy, is to be considered. PMID- 8235234 TI - [Presentation at the Salpetriere. March 1991. Changes in the general condition, intracranial hypertension and subacute meningitis in a 45-year-old man]. PMID- 8235235 TI - Review: sleep in heart failure. PMID- 8235236 TI - Cardiac failure and benzodiazepines. AB - Nine patients with stable cardiac failure and mean left ventricular ejection fraction of 30% were investigated. All had previously been prescribed a benzodiazepine hypnotic by their home physicians, but the medication had been discontinued for at least 1 month. Subjects were monitored under three conditions: 1) without any sleeping medication, 2) during nasal CPAP administration and 3) at two points during a month-long administration of the benzodiazepine that had initially been prescribed to them. Overall, the benzodiazepine hypnotic improved the sleep fragmentation noted in these patients by decreasing the arousal index from a mean of 18 +/- 6 per hour at baseline to a mean of 9 +/- 6.5 per hour after one month of benzodiazepine therapy. Total nocturnal sleep time was consequently improved [baseline mean nocturnal total sleep time: 313 +/- 27.3 minutes; benzodiazepine mean nocturnal total sleep time: 350 +/- 17.3 minutes (p < 0.0003)], as was sleep efficiency. However, the benzodiazepine hypnotic had no significant effect on central hypopneas or apneas [baseline mean respiratory disturbance index (RDI): 20.5 +/- 5.85 events/hour; mean RDI after 1 month of drug intake: 21.3 +/- 2.5 events/hour]. Nasal CPAP was also ineffective on the disordered breathing. In this group of subjects, respiration was even significantly worsened with nasal CPAP compared to baseline, as indicated by RDI (p < 0.0001), lowest SaO2 (p < 0.0001) and total nocturnal sleep time (p < 0.0001) measurements. PMID- 8235237 TI - Benzodiazepines in congestive heart failure: effects of temazepam on arousability and Cheyne-Stokes respiration. AB - We studied seven male patients with moderate to severe congestive heart failure (CHF) [left ventricular ejection fraction (LVEF) = 22.4 +/- 6.7; mean +/- SD] in a double-blind crossover trial to determine the effects of temazepam 15 mg on arousability, sleep architecture, Cheyne-Stokes respiration (CSR) and nighttime oxygen saturation. Sleep architecture was not markedly improved with temazepam. There was no significant change in total sleep time (TST) (383.1 +/- 14.1 minutes to 396.6 +/- 15.4 minutes, p = ns) (mean +/- SE, placebo vs. temazepam) or total wake time (TWT) (96.9 +/- 14.0 vs. 81.4 +/- 14.0 minutes, p = ns). Sleep stage proportions did not change appreciably except for a reduction in stage 1 sleep (6.7 +/- 1.2% vs. 4.0 +/- 1.0%, p < 0.05). Microarousals per hour of sleep decreased with temazepam (21.1 +/- 2.7/hour vs. 13.9 +/- 2.1/hour placebo, p < 0.05), with the largest change occurring in stage 2 (24.9 +/- 5.4/hour vs. 15.0 +/- 3.1/hour, p < 0.05). Wake time during sleep (WDS) was reduced from 82.5 +/- 11.7 minutes to 54.5 +/- 9.4 minutes, p < 0.03. Daytime alertness was improved with temazepam as was indicated by an increase in mean latency to sleep [multiple sleep latency test (MSLT) = 7.1 +/- 2.4 vs. 5.7 +/- 2.0 minutes, p < 0.04) on days following treatment with temazepam. There was no significant change in CSR as a percentage of TST (38.7 +/- 13.6% vs. 32.5 +/- 11.8%, p = ns). However, the apnea/hypopnea index (AHI) (10% filter) was decreased in stage 1 (28.1 +/- 9.7/hour vs. 15.6 +/- 8.2/hour). Overnight oxygen saturation did not change with temazepam (95.1 +/- 0.6% both nights) and the percentage of TST spent below 90% oxygen saturation was minimal for both conditions (1.5 +/- 1.1% vs. 2.2 +/- 1.7%, p = ns). We conclude that CHF patients with CSR experience frequent arousals and that these arousals can be reduced with temazepam. There was an improvement in daytime somnolence. There was no worsening of nighttime oxygen saturation. PMID- 8235238 TI - Effect of nasal continuous positive airway pressure during sleep on 24-hour blood pressure in obstructive sleep apnea. AB - Ambulatory blood pressure (BP) was measured noninvasively (Oxford Medilog ABP) at 15-minute intervals for 24 hours before and after 8 weeks of treatment with nasal continuous positive airway pressure (nCPAP) in 19 men with obstructive sleep apnea (OSA). We included both normotensive and hypertensive patients, but hypertensives were studied after withdrawal of antihypertensive drugs. Ambulatory BP before and after treatment was compared using patients as their own controls. Treatment with nCPAP was successfully established in 14 of the 19 patients (74%). Blood pressure fell significantly in patients who were successfully treated: 24 hour mean BP (systolic/diastolic) decreased from 141 +/- 18/89 +/- 11 mm Hg to 134 +/- 19/85 +/- 13 mm Hg (p < 0.05). The reduction in 24-hour mean systolic BP occurred during both day and night, but a significant fall in mean diastolic BP was only observed during the day. The mean blood pressure fell in both normotensive and hypertensive patients. Patients who were inadequately treated with nCPAP had no reduction in mean 24-hour BP. Effective treatment of sleep apnea with nCPAP was associated with a significant fall in both systolic and diastolic BP independent of changes in body weight or alcohol consumption, suggesting that sleep apnea was an independent factor contributing to elevated nighttime and daytime BP in these patients. PMID- 8235239 TI - Long-term nasal continuous positive airway pressure administration can normalize hypertension in obstructive sleep apnea patients. AB - We investigated the way in which nasal continuous positive airway pressure (CPAP) affects the circadian profiles of blood pressure (BP) and heart rate (HR) in obstructive sleep apnea syndrome (OSAS) patients. Nine patients with OSAS, confirmed by nocturnal polysomnography, were studied with ambulatory blood pressure monitoring (Colin ABPM-630) during two 48-hour periods, before and during nasal CPAP treatment, at the Stanford University Sleep Disorders Clinic. During each 48-hour monitoring period, blood pressure measurements were taken by the ambulatory device every 30 minutes. During the ambulatory blood pressure recordings, nocturnal sleep time was defined as the period between 0000 hours to 0600 hours and active daytime was defined as the period between 1000 hours and 2000 hours. An average systolic blood pressure > 135 mm Hg during the 48-hour baseline recording was defined as hypertensive. Using these criteria, we selected four hypertensive and five normotensive patients. Average BP (systolic/diastolic) and HR during the 48-hour periods decreased significantly from 148.6/88.2 mm Hg to 138.7/81.4 mm Hg, and from 77.9 beats per minute (bpm) to 67.2 bpm in hypertensives during CPAP treatment (p = 0.04), but there were no significant changes observed in normotensives. Average BP, during the day and at night, decreased from 152.3/91.8 mm Hg to 141.2/85.1 mm Hg and from 133.9/76.8 mm Hg to 125.9/73.7 mm Hg, respectively, in the hypertensives during CPAP, but such changes were not observed in normotensives. Average HR during the day and at night decreased significantly from 85.2 bpm to 72.6 bpm and from 69.8 bpm to 56.5 bpm in the hypertensives (p = 0.04), but not in normotensives.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235240 TI - Relationship between coronary hemodynamic changes and the phasic events of rapid eye movement sleep. AB - Previous studies in dogs showed dramatic increases in coronary blood flow associated with episodes of sinus tachycardia during rapid eye movement (REM) sleep. The present study demonstrates that 90% of these surges in heart rate and coronary flow are concentrated during periods of phasic REM sleep and only 10% in tonic REM sleep. Intensely phasic REM was distinguished from moderately phasic REM sleep by the degree of phasic eye movement. The surges were three times more frequent during intensely phasic REM than in moderately phasic REM sleep. However, the magnitudes of heart rate (37% +/- 3%) and coronary flow (25% +/- 3%) surges were unaffected by the specific substage of REM sleep. The incidence of surge events was almost eleven times greater in epochs of phasic REM that also contained a muscle twitch than in those that did not. During REM sleep, muscle twitches accompanying surges were not associated with any additional elevations in coronary flow or myocardial demand. Our data indicate that the sinus tachycardia-associated surges in coronary flow represent integrated autonomic responses intrinsic to phasic periods of REM sleep in dogs. PMID- 8235241 TI - ST segment abnormality, angina during sleep and obstructive sleep apnea. PMID- 8235242 TI - Nocturnal motor activity in a community sample. AB - The relationships between nocturnal motor activity and daytime psychophysiological activation were investigated in a random community sample of 199 subjects aged 35-55 years. Nocturnal motor activity was recorded with the static charge sensitive bed (SCSB, Bio-Matt). The association of nocturnal motor activity with demographic features, health status, laboratory blood values, afternoon electrodermal activity (EDA) and psychological distress was studied. A model for nocturnal motor activity was constructed and statistically analyzed. The analysis revealed that psychological distress, breathing disturbance, plasma glucose level and sympathetic activity were related significantly and independently to nocturnal motor activity. Their relations and the associations of sex, age, body mass index (BMI), sleep latency and health status with nocturnal motor activity were discussed in the context of the arousal theory of poor sleep. PMID- 8235243 TI - Periodic leg movements (PLM): their relationship to sleep stages. AB - We investigated the characteristics of periodic leg movements (PLM) during nocturnal sleep and wakefulness in 13 drug-free patients presenting with the restless legs syndrome (RLS, n = 9) or with isolated PLM (n = 4). Eight-hour polygraphic sleep recordings included the electromyogram (EMG) of both tibialis anterior muscles. Scoring of leg movements was done according to established criteria for periodic movements in sleep, but movements occurring during episodes of wakefulness were scored as well. Twelve out of 13 patients had PLM during wakefulness, including three subjects not affected by RLS. The frequency of periodic movements in sleep (PMS) per hour of total sleep time was significantly lower than the frequency of PLM (including movements during wakefulness) per hour of polygraphic recording. Movement indices based on PMS alone underestimated the relative frequency of PLM particularly in patients with high amounts of wakefulness (> 20%). All features of PLM clearly differed between sleep stages. Relative frequency of movements, their duration and their arousing effect decreased along the nonrapid eye movement (NREM) sleep stages, whereas the intermovement interval increased. During rapid eye movement (REM) sleep the duration of movements was shortest and the intermovement interval was longest. The results presented suggest that the processes underlying PLM are most active at the transition from wakefulness to sleep and considerably attenuated during deep NREM sleep and even more during REM sleep. We suggest including movements during wakefulness in routine PLM scoring to get a more complete picture of the disturbance. PMID- 8235244 TI - Stability and instability of sleep in older persons recorded in the home. AB - Using a nonintrusive procedure for sleep monitoring, four 24-hour recordings of 40 independently living women, 65-94 years, were made during a 3-month period. Although the subjects distributed their sleep over much of the day, all measures showed significant reliability: with four recordings, reliabilities (r44 values) were > 0.70; and with only one recording, reliabilities (r11) were > 0.40 [r11 for the respiratory disturbance index (RDI) was lower but still significant]. Within-individual variability over recordings was low for total sleep time (TST), sleep (S), and sleep efficiency (SE) and high for sleep latency (SL) and RDI. The mean TST was 7.6 hours, which is not markedly different from that of younger adults. Within the age range studied, there was no relationship between age and any of the variables, including the amount of within-individual variability. These results, using the Home Monitoring System (HMS), indicate that both stability and instability of sleep parameters characterize the sleep of older women. The nature of specific forms of individual variability needs to be explored as possible indices of aging as distinct from indications of neurobiological abnormalities. PMID- 8235245 TI - Hartley transforms and narrow bessel bandpass filters produce similar power spectra of multiple frequency oscillators and all-night EEG. AB - Frequency specific power obtained from time and frequency domain analyses are explored in simulated signals and all-night electroencephalogram (EEG). Signals were subjected to a fast Hartley transformation (FHT) and to digital sixth-order Bessel bandpass filters (BDF) of the infinite impulse response type. Numeric values of FHT, BDF and, if suited, authentic frequency specific power were subjected to a Pearson correlation. Frequency bins at 1.6-2.4 Hz (delta), 4.75 5.9 Hz (theta), 9.3-11.5 Hz (alpha), 12.5-14.9 Hz (sigma) and 16.6-19.5 Hz (beta) were investigated. When compared with true power of single frequency oscillators (256-sample windows), frequency specific power of the FHT correlated functionally (1.0) and BDF correlated highly (0.85, delta; 0.99, other bins). For analyses of "white noise", a multiple frequency oscillator and all-night EEG, four rectangular window sizes were applied (256, 512, 1,024 or 2,048 samples). The FHT power correlated better with authentic frequency specific power of "white noise" (256-sample windows) (0.61-0.98) than BDF power (0.67-0.89). With 512-sample windows of "white noise", the estimate of both the FHT (0.69-0.99) and BDF (0.71 0.93) improved. Direct comparison between FHT and BDF frequency specific power obtained from "white noise" or all-night EEG revealed a high degree of compliance between methods for all frequency bins (up to 0.99). For delta, the accord was relatively low for the 256-sample window (EEG, 0.68; "white noise", 0.72), but increased with lengthening window size (2,048-sample: 0.97; 0.99). Averaging of multiple EEG 256-sample windows also increased the agreement between methods.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235246 TI - Bibliography of recent literature in sleep research. PMID- 8235247 TI - Age and loss of bone density in pre-menopausal women. AB - Some studies have suggested that premenopausal women show a decline in bone mass in function of age while other studies have not. Bone density in the spinal column and proximal femur (neck, Ward's triangle and trochanter) in 75 healthy white women between the ages of 30 and 49 without a history of irregular menstrual periods was correlated with age, weight, height and body mass. No statistically significant decline in bone density with age was seen during the premenopausal period. There was a positive correlation between bone density in the spine and proximal femur and weight and body mass indices. Our results suggest that healthy women do not show loss before menopause and it also seems that obesity has a favorable effect on these women's bone mass. PMID- 8235248 TI - Retrospective prevalence and incidence of drug-induced agranulocytosis in the city of Sao Paulo-Brasil. AB - Retrospective study carried out in Sao Paulo, Brazil, from September 1981 through March 1990. During this period the study investigated 19,389 hematological patients, aiming to assess the prevalence and estimate the incidence of drug induced agranulocytosis. To assess the Hematology Center's catch population, where the study took place, we adopted the incidence of hemophilia A, chronic myeloid leukemia and acute leukemia, described by the international literature as reference for our catch population estimate. Our findings revealed a prevalence of 1 case of agranulocytosis per 3,878 subjects treated at the Sao Paulo Hematology Center which translates into an incidence of 0.44 to 0.82 cases of agranulocytosis per million inhabitants, per year. PMID- 8235249 TI - Study in necropsy material of "cause-specific mortality" in diabetics, in Sao Paulo-Brasil. AB - Specific causes of death in diabetic patients was studied in retrospective work from necropsies performed at the Medicine School of the University of Sao Paulo from 1931 until 1989. From 145,043 necropsies, 813 indexed cases were found: 449 females (55.2%) and 364 males (44.8%). The race ratio of patients was: 584 (71.8%) white, 208 (25.6%) negro and 21 (2.6%) asiatic. The predominant age of death incidence was between the fifth and sixth decades. The main causes reported were infections (42.68%), coma (12.79%), neurologic involvement (11.56%) and cardiac diseases (11.07%); indeterminate causes of death were related in 7.4% of cases. Our results differ from the others of literature mainly in two points: the high prevalence of infectious diseases and the proportional low incidence of cardiopathies. We concluded that, in our casuistic, infectious diseases in diabetic patients represent an important cause of death establishing important clinical parameters for their care. PMID- 8235250 TI - Nipple discharge--study on 100 patients. AB - In a retrospective study the authors analyzed 100 cases of women with serous, serosanguineous, and sanguineous nipple discharges, without palpable and radiologically visible tumors, that were submitted to resection of main galactophorous ducts and biopsy of underlying breast parenchyma. The sensibility of the cytologic examination of nipple discharges for carcinomas was 16.6%. The histopathology showed incidence of 61% of mammary dysplasia, 33% of intraductal papillomas single or multiple and 6% of carcinomas. The early diagnosis of carcinoma gave good results after radical surgical treatment. PMID- 8235251 TI - Anthropometric nutritional assessment of critically ill hospitalized children. AB - In a prospective study, the nutritional status was evaluated in 46 consecutive admissions to a Pediatric Intensive Care Unit, using anthropometric parameters. About 65% of the patients presented malnutrition on admission, with predominance of the chronic form. The mortality rate was greater in the malnourished (20 versus 12.5%) and chronic malnutrition was associated with a higher incidence of infection (42%). There was a fall in channel of percentile for weight-for-height in 36% of the patients evaluated at the final follow up. These results suggest that a significant number of patients are at nutritional risk at the time of hospital admission, and there is an association between nutritional status and hospital course. The anthropometric nutritional evaluation is a simple an reproducible method, and a valuable parameter for an objective nutritional assessment of the critically ill child. Therefore, it should be performed on admission and follow up of hospitalized children. PMID- 8235252 TI - Cartilage-hair hypoplasia syndrome: immunological evaluation of two cases. AB - PURPOSE: Immunological evaluation of patients with cartilage-hair hypoplasia. TYPE: Prospective and retrospective studies. PLACE: Division of Allergy, Clinical Immunology and Rheumatology-Dept. of Pediatrics-"Escola Paulista de Medicina". PATIENTS: Two children with cartilage-hair hypoplasia syndrome. METHODS: Clinical and immunological evaluation. Humoral immunity (immunoglobulin levels, poliovirus antibodies, etc.) and T cell immunity (in vitro cultured lymphocytes stimulated with PHA, Con A and PWN, total T cell and subset determination) were studied. RESULTS: Cellular immunodeficiency and hypogammaglobulinemia were observed in one patient and normal values in the other. CONCLUSIONS: Immunological evaluation (cellular and humoral) should be performed in all patients with cartilage-hair hypoplasia. PMID- 8235253 TI - Respiratory monitoring: pulse oximetry and capnography in children during anesthesia and intensive care. AB - Monitoring the critical care patient by pulse oximetry and capnography permits the early diagnosis and follow-up of various clinical events in a precise manner, with considerable safety and with no need for invasive procedures. These techniques have been extensively evaluated in adults, but only recently have they been studied in pediatric patients, including newborn infants. In the present review we discuss the working principles, indications, advantages and limitations of each technique, as well as the interpretation of the results. PMID- 8235254 TI - Cytology by capillarity and anatomopathological diagnosis of solid breast tumors. PMID- 8235255 TI - Cytology by capillary. PMID- 8235256 TI - [Indications and limitations of non-hormonal treatments of osteoporosis]. AB - Postmenopausal osteoporosis is a multifactorial disease stemming from the accumulation of several risk factors, among which estrogen deficiency plays a key role. Although the only currently available preventive treatment is replacement estrogen and progesterone therapy, there is evidence suggesting that calcium influences cortical bone loss. In contrast, calcium alone has no effect on trabecular bone loss. Calcitonin, biphosphonates and fluoride salts are not indicated for primary prevention, in patients with no fractures. Ongoing studies are evaluating the efficacy of intranasal calcitonin and of biphosphonates for the prevention of postmenopausal osteoporosis. In the future, these specific treatments will be indicated only in women at high risk for osteoporosis. Curative therapy of patients with trabecular osteoporosis responsible for vertebral crush fractures rests on fluoride salts, calcitonin and etidronate. Fluoride is the only compound capable of inducing a substantial rise in bone mass. Some studies, but not all, suggest that fluoride treatment reduces the fracture risk. Determining the effective dosage is essential. Calcitonin is used in France in sequential treatment. Although sequentially administered calcitonin may increase bone mass, there is no other evidence that this drug reduces the fracture risk. Disodium etidronate given sequentially has recently been approved for use in osteoporosis because of data demonstrating that it decreases the incidence of new fractures over a two-year period. PMID- 8235257 TI - [Respective importance of the different risk factors for osteoporosis]. AB - Among the factors which influence accumulation of bone mass, genetic factors related to race and family are preponderant: they account for approximately 80% of the variance observed in adulthood. At puberty, nutritional factors and physical activity modulate the accumulation of bone and determine whether or not optimal bone mass is achieved. In women, estrogen deprivation following menopause, and also more minor disorders of gonad function, are the main causes of decreased bone mass; their respective contributions are analyzed. PMID- 8235258 TI - [Substitutive hormone therapy in postmenopausal osteoporosis. Are there any contraindications?]. AB - Estrogen deficiency is the main cause of postmenopausal osteoporosis. Replacement estrogen therapy protects the bone by reducing bone resorption and activating osteoblasts, as well as by promoting absorption of calcium and production of calcitonin. These preventive effects are especially marked in cancellous bone, provided estrogen therapy is initiated as soon as menstruation stops. Effects are dose-dependent and efficacy of the treatment is noticeable mainly during the period of administration. The rare contra-indications to estrogen therapy include hormone-dependent cancers, cholestatic jaundice, and large uterine myomas (for which surgical treatment is recommended). Metabolic disorders, arterial hypertension, and thyroid function disorders are less common with the new natural estrogens (estradiol) given orally or percutaneously to avoid hepatic passage of the drug. As for treatment induced cancers, sequential administration of a progestagen protects the endometrium and the relative risk seems negligible for breast cancer. Although concomitant use of a progestagen is mandatory, either natural progesterone or norpregnanes should be given to avoid adverse metabolic effects. Emphasis has recently been put on the role of concomitant progestagen therapy which may promote the formation of bone, probably by competing for glucocorticoid receptors in bone. There is still a need for prospective epidemiological studies, although evaluation methods and the long follow-ups needed raise significant problems. PMID- 8235259 TI - [Contribution of medical imaging. What examinations for what indications?]. AB - Osteoporosis is an obvious diagnosis in patients with fractures due to decreased bone mass. However, in many cases, major bone loss is infraclinical and requires quantification. Two methods are available to achieve this, i.e., dual energy X ray absorptiometry (DXA) and quantitative computerized tomography (QCT). QCT, which measures the central trabeculae in the lumbar vertebra, is a sensitive technique and has demonstrated a mean bone loss of 1.1 p. cent per year in cancellous bone. However, use of QCT is limited by the high radiation exposure and costs involved. Dual energy X ray absorptiometry is a highly reproducible method which ensures satisfactory correction of soft tissue effects and entails only low exposures to radiation. However, errors occur in patients with aortic calcifications, posteriorly located osteophytes or scoliosis. The optimal frequency of quantitative evaluations of bone varies with the type of disease; in patients with osteoporosis, it is unnecessary to perform evaluations more than once every three years to monitor the course of the disease. PMID- 8235260 TI - [A case of cervico-isthmic pregnancy with delayed development]. AB - This is the case of a para three, gravida three, who had a caesarean section for her first pregnancy and a normal delivery for the second one. During her third pregnancy the patient is hospitalised for first trimester bleeding. Ultrasounds show two liquid compartments separated by placenta. The lower one in cervico isthmic position contains the foetus. During the 24th week gestation the pregnancy is complicated by severe metrorrhagia that leads to abdominal exploration and total hysterectomy. The pathology conclusions confirmed the per operative impression of cervico-isthmic pregnancy. The authors emphasise the rarity of the cervico-isthmic pregnancy (1 for 10,000 pregnancies) and wonder about atypical evolution of this case and about the possibilities of saving the uterus by an earlier intervention in view of ultrasonographic data. PMID- 8235261 TI - [Value of standardized Ginkgo biloba extract (EGb 761) in the management of congestive symptoms of premenstrual syndrome]. AB - The efficacy of standardized Ginkgo biloba extract (EGb 761) in treating congestive symptoms of premenstrual syndrome (PMS) was evaluated in a controlled multicentric double blind study versus placebo. The population studied was a group of 165 women aged between 18 to 45, in genital activity period, suffering since 3 cycles from congestive premenstrual troubles during at least 7 days per cycle. The characteristics of patients and PMS were the same in both groups (EGb 761 and placebo). The observation of one menstrual cycle confirmed the diagnosis of PMS. Then, during the 2 following cycles, each patient received either EGb 761 or placebo from the 16th day of the first cycle till the 5th day of the next cycle. A double evaluation of the symptoms was realized by the patient using a daily rating scale (auto-evaluation), by the practitioner during visits at the premenstrual phase before and after the two cycles treatment. From 165 patients included, 143 observations were available. With a good acceptability, EGb 761 was effective against the congestive symptoms of PMS, particularly breast symptoms with a statistical significance between EGb 761 and placebo. Neuropsychological symptoms were also improved. EGb 761 is an alternative of interest to therapeutics already used in treating PMS or can be associated without any inconvenience. PMID- 8235262 TI - [New perspectives in the treatment of rheumatoid disease]. PMID- 8235263 TI - Arrangement of the collagen and elastic fibers in the upper human duodenum. AB - The arrangement of the elastic and collagen fibers in the upper human duodenum was examined. These fibers lie parallel to the muscle fibers in the tunica muscularis. In the submucosa they form a pantographic network arrangement, the cross-over angles of which increase in the oro-aboral direction. The collagen and elastic fibers show a polar orientation among the muscle fibers. PMID- 8235264 TI - [Vascular complications of the Ilizarov method]. AB - Between 1987 and 1991, 209 patients were operated by the Ilizarov method in the Department of Orthopaedic and Traumatologic Surgery of the University of Sao Paulo Medical School. Five of them presented vascular complications during the treatment. All were adults treated for long bone pseudarthrosis. The vascular lesion occurred in the femoral superficial artery (one case), in the popliteal artery (one case), and in the posterior tibial artery (one case); in the remaining two patients it was not possible to determine the exact site of the lesion. The clinical manifestation of the vascular problem was bleeding at the cutaneous point of the Kirschner transfixation wires. Arteriography was done in all cases but its interpretation was difficult because of the presence of the external fixator. The treatment of the vascular complications was following: in the case of injury in the femoral artery, the external fixator was taken off and a by-pass with an inverted graft of the saphenous vein was made the same procedure was done in the popliteal artery but in this case it was not necessary to remove the external fixator; the remaining case with the lesion in the posterior tibial artery was treated by ligature. In two cases where the artery involved was not determined--one in the upper extremity with the external fixator applied to the femur--the bleeding stopped after the removal of wires. PMID- 8235265 TI - Thymectomy in over 50 years-old patients with myasthenia gravis: long and short term results in 21 patients. AB - Thymectomy is considered to be an important method for the treatment of myasthenia gravis in older patients when the improvement cannot be achieved with the use certain drugs such as anticholinesterasic, corticosteroids and immunosuppressive agents. In our series of 281 thymectomized patients, 13 were male and 8 female, varying their age between 50 and 73 years. The duration of the disease prior to the thymectomy varied between 30 days and 15 years; in 12 of them (57%) it lasted already 2 years. Eleven patients had the severe and eight the moderate clinical form of the disease. Nine patients were treated with corticosteroids before submitted to thymectomy. In 11 patients the histopathological changes in the thymus were evaluated: eight had thymomas, one atrophic changes, one lipomatosis and one a normal thymus. The short-term follow up revealed important improvement in 12 patients, slight improvement in four and worsening in two; two patients died. It is concluded that thymectomy may be indicated in older patients when their condition worsens in spite of the use of corticosteroids and immunosuppressive agents, in the presence of serological markers or computerized tomography image indicating thymoma, and intolerance for corticosteroids. PMID- 8235266 TI - [Duodenal perforation following endoscopic papillotomy: report of 3 cases]. AB - Duodenal perforation is a rare complication of endoscopic sphincterotomy and its treatment is controversial. The authors report three cases of duodenal perforation by endoscopic sphincterotomy. In two cases with history of a previous biliary infection, surgical treatment had to been performed because of a peripancreatic abscess. PMID- 8235267 TI - [Frantz's tumor (papillary epithelial and cystic neoplasms of the pancreas): report of 3 cases]. AB - Three cases of Frantz tumor, a papillary and cystic tumor of the pancreas, are discussed and compared with other cases reported in the literature. The neoplasm behaved in these three cases as a benign tumor. Pancreaticoduodenectomy was performed in one patient and distal pancreatectomy in the other two. C T scan and ultrasonography are useful for the diagnosis of these tumors. Cut surfaces show mostly well encapsulated tumors with the characteristically solid and hemorrhagic necrotic patterns. Usually they are of a very low-grade malignancy so the complete surgical removal is the treatment of choice. PMID- 8235268 TI - [Traumatic right diaphragmatic hernia: report of a case and review of the diagnostic methods]. AB - Rupture of the right hemidiaphragm from blunt trauma is uncommon. Better management and more rapid transportation of victims has increased the frequency with which patients sustaining this injury are arriving at the emergency room alive. A case of right-sided diaphragmatic hernia from blunt trauma in a 36-year old man is reported. Diagnosis was established by chest x-ray, CT scan and digital exploration after unsuccessful thoracoscopy. The methods used for diagnosis of this condition are analysed. PMID- 8235269 TI - [Tissue necrosis: a side effect of sodium diclofenac: report of cases and discussion of the physiopathology]. AB - Diclofenac sodium is a non-steroidal anti-inflammatory widely used drug both in the treatment of the rheumatic diseases and as analgesic. The fact that is a potent analgesic spread its use to almost all fields in medicine, increasing the reports of adverse effects associated with it. The main complications reported are gastrointestinal, dermatological and of the central nervous system. The authors present seven cases that developed tissue necrosis as a result of the intramuscular injection of diclofenac sodium. Because of the low incidence or this adverse effect, a review is carried out and the physiopathologic mechanisms likely to be involved are discussed. Our aim is to warn against the indiscriminate intramuscular use of this drug. PMID- 8235270 TI - [The laparoscopy method in preoperative evaluation of patients with malignant stomach neoplasm]. AB - In 190 patients with malignant gastric neoplasia the value of laparoscopy for diagnosing intra-abdominal metastases was investigated. At the laparoscopic examination the presence of tumoral tissue was appraised on the serosal coat of the stomach, epiploons, parietal and visceral peritoneum, and in the liver. On hand of laparoscopic findings (peritoneal or hepatic metastases) the surgical treatment was discarded in 34 patients (70.6%). In the remaining 29.4% of patients a bad general condition or concomitant diseases contra-indicated surgical intervention. A comparison of laparoscopic and surgical findings showed that correct diagnosis was established by laparoscopy in 61.4% of cases with tumoral invasion of gastric serosa, in 21.4% of cases with involvement of epiploon, in 27% of parietal peritoneum, and in 33.3% of the liver. Falsely positive findings were rare (5% of examinations). PMID- 8235271 TI - [Superposition syndrome of connective tissue diseases: current view with special focus on mixed connective tissue disease]. AB - Attention is called to mixed connective tissue disease which, twenty years after it's original description, has now reached the syndromic individualization with important therapeutic and prognostic implications. In particular the discussion concerns the pulmonary complications (hypertension and fibrosis) responsible frequently for fatal outcome. PMID- 8235272 TI - [Preference for general practice in the medical course: relationship with changes in the clinical environment]. AB - Career preferences were investigated before and after a major change of clinical training setting at the University of Brasilia. The inquiry addressed to pre clinical students for a total of 15 cohort years (N = 1107). Data were gathered through self-report questionnaires on main preference, its inducing factor (a priori, theme study, task experience) and on learning style. The results showed a significant difference between student-pool career preference profiles, before and after change, which was mostly due to a decrease (from 24% to 10%) in preference for general practice. Lesser proportions of aprioristic induction, active learning style and male students were featured in the after-change student pool. Relation-ships were found between preference for general practice and aprioristic induction, active learning style and male sex. Decline in preference for general practice is ascribed more to factors reducing access to course than to reorientation of the curriculum. PMID- 8235273 TI - [Infection involving arterial prosthesis: clinical picture, etiology, and predisposing factors]. AB - Thirty patients were operated for arterial reconstruction with synthetic grafts. They presented one of the following complications: 1) prosthesis exposed by a cutaneous fistula; 2) prosthesis in communication with a hollow viscus; 3) prosthesis involved by pus; 4) positive culture of a fragment of the prosthesis or of the surrounding secretion. The clinical manifestations were cutaneous fistula in 21 patients (70%), external bleeding in 14 (47%), exposition of the prosthesis in five (17%), anastomotic aneurysm in five (17%), and enteric fistula in four (13%). The infection became evident during the first postoperative year in half of the patients. The most important agents of infection were staphyilococci and Gram negative bacteria. PMID- 8235274 TI - [Acute nephrotoxicity caused by amphotericin B in the dog]. AB - Amphotericin-B is the drug of choice for the treatment of serious systemic fungal infections. The major limitation for the use of this drug is its potential nephrotoxicity. The chronic and acute administration causes both a reduction of renal blood flow and glomerular filtration rate, determining a fall of renal concentrating ability. Our study was designed to determine the acute effects of Amphotericin-B on dogs renal function, during hydropenia and water diuresis. In addition a hemodynamic study, with the measurement of intrarenal blood flow by the microsphere method was performed. The hydropenic dogs received the drug directly into the left renal artery. There was a significant decrease in creatinine clearance, urinary osmolality, and urinary solute excretion, in the left kidney only. Amphotericin-B was infused into the aorta above the renal arteries in the group in which the water diuresis was induced. This acute infusion produced intense fall in creatinine clearance and in the free water formation, as well as in absolute excretion of sodium. The hemodynamic study showed that the reduction of creatinine clearance may be due to the decrease in renal blood flow, and the alterations in the urinary excretion of water and sodium could not be correlated with the distribution of cortical blood flow to the deep nephrons. These findings suggest that the effects of Amphotericin-B may be attributed, at least in part, to its ischemic action. PMID- 8235275 TI - [Cavity effusions in patients with chronic pancreatitis]. AB - In Brazil, the incidence of IPF due to alcohol-induced CP seems to be higher than in other countries. The authors analysed some general, epidemiologic, laboratory and radiologic features in 98 patients with CP, all of them alcoholics, divided in two groups: 49 patients without IPF (I) and 49 patients with IPF (II). The authors first analysed the following parameters in each group and then comparatively in both groups: age, sex, race, smokers and daily consumption of cigarettes, daily consumption of alcoholic beverages, years of alcohol consumption, and mortality. The authors concluded that pancreatic fistula plays in effusions the main role, secondary to cysts disruption or necrosis of the main pancreatic duct. PMID- 8235276 TI - [Local treatment of burns: analysis of the use of antimicrobial agents]. AB - The authors studied the local treatment of burns in 11045 cases: 6063 (54.9%) were treated without topical antibacterials and 4985 (45.1%) with antibacterial agents. In 62.1% of cases the topical antibacterial treatment was done mainly in the hospital ward (69.5%) while the treatment without topical medication was mostly performed in the operating room. The use of topical antibacterials was studied during eight years, from 1985 to 1992. In the earlier periods the justifications were not related to burn wound sepsis. In the recent years the topical antibacterials were used chiefly for the treatment and prophylaxis of wound infection (96.8% of cases). PMID- 8235277 TI - [Surgical treatment of infections involving arterial prosthesis in aorto-femoral position]. AB - This article is to evaluate the results of surgical treatment of aorto-femoral graft infections. The records of 20 patients with aorto-femoral graft infections were reviewed. The patients were submitted mostly to a combination of surgical procedures. They were: a) conservative treatment; b) partial removal of the infected graft and c) total removal of the infected graft. The removal of the prosthesis, partial or total, was followed by a new arterial reconstruction in some patients. The final treatment was: total removal of the infected graft in 11 (55%) patients, total removal plus arterial reconstruction in six, partial removal in two and partial removal plus arterial reconstruction in one. The results of treatment were presented as the survival rate and the effective palliation (patient alive, lower limbs preserved and infection cured). The survival rates after one month, 12, 24, 36 and 48 months were respectively: 80%, 60%, 53%, 42% and 27%. The effective palliation rates after one month and after 48 months were respectively 50% and 27%. The highest effective palliation rate was obtained when the infected prosthesis was removed and a new arterial reconstruction performed. We conclude that the treatment of choice of aorto femoral graft infections seems to be the total removal of the graft followed by new arterial reconstruction. PMID- 8235278 TI - [Traumatic rupture of the diaphragm: study of 35 cases]. AB - A retrospective review of 35 patients with diaphragmatic injury attended at the Emergency Service of the Hospital das Clinicas, University Sao Paulo is analysed. Diagnosis was made through chest roentgenogram (48.5%), peritoneal lavage (37%) and contrast studies (14.5%). The most common extra-abdominal lesion were due to head, pelvic and thoracic trauma. Thirty-one patients sustained injuries to the left-side and four to the right. Associated intra-abdominal injury occurred in 89% of them. Retroperitoneal haematoma, spleen and liver injury were the most frequent lesions. All cases were treated by laparotomy, pleural space irrigation through the diaphragmatic lesion, thoracic drainage and suture of the injury. The morbidity rate was 40% and deaths occurred in 22.8%. Based upon these data, we concluded: the mechanism of trauma, chest RX findings and pelvic fracture are important clues for early recognition; irrigation of pleural space seems to be important in order to prevent empyema. Despite of this, the mortality remains high. PMID- 8235279 TI - [Femoral hernia: anatomical basis for its predominant occurrence on the right side]. AB - The possible relationship between the greater incidence of femoral hernia on the right side of the human body and anatomic differences between the right and left femoral ring were studied. The morphology and the transverse diameter of the femoral rings on both sides of 38 adult, formolized corpses were compared. There was no difference between the two sides. PMID- 8235280 TI - [Radiological staging index of osteoarthrosis]. AB - The association of joint pain with radiological changes (such as osteophytes, joint space narrowing, and sometimes, bone cysts and erosion) is the most commonly criteria used in daily medical practice for the diagnosis of osteoarthrosis. However, these findings do not allow objective quantification of the intensity of the joint involvement. In an attempt to measure quantitatively the radiological lesions encountered in osteoarthrosis the authors elaborated a radiological index based on the following parameters: I. severity of radiological changes, II. area of the affected joint and III. number of affected joints. The application of this index in the medical practice allows a simple way of quantification of radiological lesions of osteoarthrosis and thus, a better clinical monitorization of the disease. PMID- 8235281 TI - [Paracoccidioidomycosis of the central nervous system: study of 5 cases by magnetic resonance]. AB - Paracoccidioidomycosis is a systemic mycosis, endemic in South and Central America, that affects the central nervous system in almost 10% of the cases. Neurological involvement includes to different clinical forms: meningeal and granulomatous form, also known as pseudotummoral form. Five patients with intracranial paracoccidioidomycosis were submitted to MR examination in a 1.5T system. All patients presented multiple round or lobulated lesions, hyperintense in T1-weighted images and predominantly hypointese in T2-weighted images. The lesions were distributed diffusely, with a slight predominance in the supratentorial compartment, although infratentorial lesions were also observed, mainly in the cerebellum. All lesions had ring or nodular contrast enhancement. Three patients were reexamined for treatment control, with disappearance of the lesions in two, with persistence of hypointense area in T2-weighted images. MR imaging proves to be a good method to evaluate the neurological involvement in paracoccidioidomycosis, mainly due to its high sensitivity for posterior fossa lesions. Further studies are needed to correlate the MR findings and the changes found in MR examination after treatment. PMID- 8235282 TI - [Gastric epithelioid leiomyosarcoma simulating pancreatic pseudocyst. Report of a case and review of the literature]. AB - The authors report a case of malignant gastric leiomyoblastoma (epithelioid leiomyosarcoma) which was mistaken for a pseudocyst of the pancreas. The patient was operated several times in another hospital. At one of the operations a cystogastrostomy was performed. In november, 1989, when operated again by us, a large tumoral mass arising from the posterior gastric wall was found. The histopathological diagnosis was leiomyoblastoma of the stomach. It is concluded that the wall of a pancreatic cystic lesion should always be submitted to histologic examination in order to be excluded the possibility of a malignant neoplasm. PMID- 8235283 TI - Implant terminology survey results. PMID- 8235284 TI - Where are we? PMID- 8235285 TI - An esthetic solution to a difficult implant restoration. PMID- 8235286 TI - Laboratory procedures for fabrication of a self-curing resin reduction coping and intraoral application. PMID- 8235287 TI - Infection control in the dental laboratory. PMID- 8235288 TI - Long-term experience with low dose methotrexate in rheumatoid arthritis. AB - One hundred twenty-six patients with rheumatoid arthritis (RA) were treated with weekly low doses of methotrexate (MTX) for a mean period of 36.8 months (range 13 110 months). The overall probability of continuing with MTX therapy was 72% at 2 and 3 years, 67% at 4 years and 65% at 5-7 years. Seronegative patients had a higher probability of continuing therapy than seropositive patients (P < 0.05). Out of the whole group, 8% showed no improvement, 16% showed mild improvement, 30% showed moderate improvement, and 45% experienced marked improvement. Eight patients (6%) of the latter group achieved complete clinical remission. In the course of the follow-up period there was a significant decrease in the mean daily dosage of prednisone and NSAIDs. Minor side effects were common (68%), but therapy was discontinued in only 27 patients (21%) because of major complications. In most of them (25 out of 27) these occurred within the first 24 months of therapy. Although malignancy was revealed in 5 patients during the follow-up period, its occurrence did not differ from expected rates. PMID- 8235289 TI - Differences in anti-Fab antibodies in adult and late onset rheumatoid arthritis. AB - In the present study the ratio of antigen-bound anti-IgG-Fab antibodies (hidden aFab) to free aFab was found to be significantly increased in patients with adult onset rheumatoid arthritis (AORA) as compared to late onset rheumatoid arthritis (LORA). The overall amount of aFab was similar in both groups. The difference was only seen in seropositive patients. Within the seropositive AORA group, the aFab ratio was correlated with the duration and the stage of disease but not with the patients' age at investigation. This might reflect a higher affinity of anti-Fab antibodies and/or a greater diversity of the idiotypic repertoire in adult onset disease resulting in the formation of immune complexes, the stability of which might be enhanced further by the presence of rheumatoid factors. Although a pathophysiological involvement of aFab cannot be concluded from our observations, it is conceivable that different immunoregulatory mechanisms could be operative in RA with onset at different ages. PMID- 8235290 TI - Levels of soluble receptors for tumor necrosis factor type I and type II in paired synovial fluids of arthritis patients. AB - The objective of this study was to determine whether levels of soluble receptors for tumor necrosis factor type I and type II (sTNF-RI and sTNF-RII) as measured in paired synovial fluids (SF) of arthritis patients are associated with clinical or laboratory parameters of local inflammation. sTNF-RI and -RII were measured by ELISA. We found that sTNF-RI and -RII did not correlate with activity of local inflammation. sTNF-RI levels correlated with sTNF-RII concentrations. We concluded that sTNF-RI and -II did not represent markers for local disease activity in arthritis patients. PMID- 8235291 TI - Immunoglobulin G and serum albumin isolated from the articular cartilage of patients with rheumatoid arthritis or osteoarthritis contain covalent heteropolymers with proteoglycans. AB - The present study was undertaken to identify the cartilage matrix molecules that are bound with intermolecular disulfide bonds to IgG and serum albumin molecules recovered from the articular cartilage of patients with rheumatoid arthritis (RA) or osteoarthritis (OA). The cartilage specimens were extracted sequentially with three changes of neutral buffer, three changes of 6 M guanidine hydrochloride and then partially degraded with bacterial collagenase. The extracted IgG and albumin, along with matrix molecules bound to these proteins, were isolated with affinity chromatography using antibodies to IgG or human serum albumin conjugated to agarose beads. The isolated materials were characterized with sodium dodecyl sulfate polyacrylamide gel electrophoresis and transfer blotting, using specific antibodies to IgG, albumin, and proteoglycans. In the isolated materials, heteropolymers with IgG or albumin were identified. These polymers contained keratan sulfate and less frequently chondroitin-4-sulfate and chondroitin-6 sulfate. These findings identified the keratan sulfate rich proteoglycans, prevalent at the surface of joint cartilage, as the most common cartilage matrix molecules that are covalently bound to IgG or to serum albumin by disulfide bonds in the articular cartilage of patients with RA or OA. PMID- 8235292 TI - Studies on the expression of the TNF alpha receptors (p55 and p75) and their relative contributions to prostanoid production and glycolytic rate by rheumatoid synovial fibroblasts in vitro. AB - The effect of monoclonal antibodies against the 55- and 75-kDa (p55 and p75) tumour necrosis factor (TNF) receptors on two tumour necrosis factor alpha (TNF alpha) induced responses was studied in rheumatoid synovial fibroblasts (RSF) in vitro. This provided functional evidence for the expression of both receptor types, which was confirmed by substantial inhibition of 125I-TNF alpha binding to RSF by both antibodies. TNF alpha stimulation of prostanoid production was found to be a function of ligand binding to both receptor types, whereas the stimulation of cellular glycolysis was largely mediated via the p55 receptor. Thus, we showed that RSF express both types of TNF receptor and functional studies showed that stimulation of each receptor results in both similar and dissimilar cellular responses. PMID- 8235293 TI - Reactive nitrogen intermediates, antinuclear antibodies and copper-thionein in serum of patients with rheumatic diseases. AB - Sera from 354 patients with various inflammatory and autoimmune rheumatic diseases were screened for the presence of reactive nitrogen intermediates, antinuclear antibodies and the anti-oxidase copper-thionein (Cu-thionein), and compared to sera from healthy donors and patients with non-rheumatic diseases including AIDS, various internal as well as neurological diseases and carcinoma of different organs. When compared to healthy individuals, the levels of nitric oxides in sera from patients with autoimmune rheumatic diseases were elevated by 240-600% (P < 0.01). The status of reactive nitrogen intermediates (NOx, RNI) in sera from donors with inflammatory rheumatic diseases was increased by 170-540%, but was also significantly enhanced in sera of patients with non-rheumatic diseases, indicating a general inflammatory mechanism that is predominantly triggered by inducible nitric oxide (NO) syntheses of phagocytes. All rheumatic sera were dramatically depleted of the anti-oxidase Cu-thionein (P < 0.001), a powerful consumer of hydroxyl radicals and singlet oxygen and an efficient superoxide dismutase. The NOx levels were positively correlated with the serum titers of antinuclear antibodies (r = 0.77) and negatively correlated with Cu thionein levels (r = 0.94), reflecting a high steady-state concentration of free radicals generated during inflammatory and autoimmune rheumatic diseases. PMID- 8235294 TI - [The microeconomic reform of health]. PMID- 8235295 TI - [The child with chronic problems. Continuing nursing care]. PMID- 8235296 TI - [Monitoring. Electronic surveillance]. PMID- 8235297 TI - [Addictions and learning processes]. PMID- 8235298 TI - [The coming meeting of the IV Congress of Catalan Nursing. The extensive debate on the situation of nursing]. PMID- 8235299 TI - [The sleep apnea syndrome in Alava. A research project on its prevalence is being conducted at Hospital Txagorritxu]. PMID- 8235300 TI - [Continuing education and quality of life]. PMID- 8235301 TI - [The cancer patient]. PMID- 8235302 TI - [The treatment of pain. 2. The pharmacology of analgesics and postoperative pain]. PMID- 8235304 TI - [The future of health in Europe]. PMID- 8235303 TI - [Absorbents for urinary incontinence in adults]. PMID- 8235305 TI - [Continuing education. 82. Medical-surgical. Cardiorespiratory arrest]. PMID- 8235306 TI - Characterization of recombinant human neuron-specific enolase and its application to enzyme immunoassay. AB - Human gamma-enolase cDNA prepared by reverse transcriptase-polymerase chain reaction was cloned into the Escherichia coli expression vector pKK223-3. The resulting plasmid, pHTK503, expressed human gamma-enolase as a 46-kDa protein in SDS-PAGE, and in the cells as the active gamma gamma form (designated as recombinant human NSE; R-NSE). R-NSE was purified from E. coli by several chromatographic elutions. Finally, 6.0 mg of R-NSE from 8.1 g cells was purified with a specific activity of 86 units/mg protein. The structural properties of R NSE were compared with the NSE purified from human brain tissue (B-NSE). The biochemical and enzymatic characteristics were essentially the same, except for the isoelectric point (4.5 for B-NSE and 4.7 for R-NSE). In an NSE immunoassay system, R-NSE and standard NSE were almost equal in reactivity to the anti-NSE antibody. These results indicate that R-NSE can be used as standard assay material. PMID- 8235307 TI - Alteration of laminin production in small-cell lung carcinoma: possible correlation with the absence of the basement membrane. AB - Basement membranes are frequently absent in small-cell lung cancer (SCLC). To study this phenomenon, the production of laminin by SCLC cells, both in vitro and in vivo, has been investigated and compared with the laminin production by lung carcinomas of other histotypes (non-SCLC, NSCLC). Immunoradiometric and immunoperoxidase tests, respectively, carried out on culture supernatants and cells using antilaminin rabbit antiserum, revealed that in 1 (H446) out of 8 SCLC cell lines tested and in the NSCLC line Calu3, laminin was detectable both in the culture medium and in the cytoplasm of the cells. After treatment of an SCLC negative cell line (N592) with monensin, a molecule which inhibits protein secretion, laminin became detectable in the cytoplasm. Similar results were obtained by FACS analysis on cells permeabilized with saponin. Northern analysis indicated that the laminin B1 gene was transcribed. The level of mRNA for the B1 laminin subunit in the N592 cells was twice and 4 times higher than that found in the laminin-secreting Calu3 and H446 cell lines. The production of laminin in SCLC and NSCLC surgically resected samples using immunoperoxidase staining of cryostatic sections was also investigated. The results indicate that 85% of the NSCLC cases tested showed diffuse staining in the cytoplasm of the tumor cells and strong staining at the basement membrane level, whereas a similar staining was only found in 15% of the SCLC cases tested. The treatment of SCLC cells with differentiating agents in vitro has been shown to induce the adhesion of these cells. In fact, n-butyric acid induced the disaggregation of floating-clump cells and single-cell adhesion in the N592 line, whereas after treatment with retinoids the clumps of the N592 cells were still present, but 30% of these were found to adhere to the plate. However, no increase in the laminin production was found in the cytoplasmic or culture medium under these conditions. PMID- 8235308 TI - Cytotoxic effect of the protein-doxorubicin conjugates on the multidrug-resistant human myelogenous leukemia cell line, K562, in vitro. AB - In vitro studies were performed to examine the antitumor effect of protein doxorubicin (DXR) conjugate on the growth of the multidrug-resistant human chronic myelogenous leukemia cell line, K562/DXR. The 50% inhibitory concentration (IC50) for DXR in the K562/DXR cell line was 20 nM (in the K562 parental cell line, IC50 was 3.2 nM). Treatment of both types of cells with various concentrations of DXR or conjugates at equivalent concentrations of DXR was carried out. One type of the conjugates used was human serum albumin (HSA) DXR conjugate and human transferrin (Tf)-DXR conjugate via a glutaraldehyde bridge (HSA-ga-DXR, Tf-ga-DXR, respectively) and another type used was HSA-DXR conjugate with a dextran bridge (HSA-dex-DXR). All of these conjugates showed potent dose-dependent inhibition of cell growth against the K562/DXR cells as compared with the cells treated with DXR or other controls. IC50 for HSA-ga-DXR, Tf-ga-DXR and HSA-dex-DXR conjugates in the K562/DXR cell line was 2.4, 3.6 and 1.0 (equivalent DXR) nM, respectively, which were approximately similar to the value of the K562 treated with DXR. Through the treatment of K562/DXR cells with HSA-DXR conjugate, the intracellular drug concentration increased as a function of time up to 24 h compared with the cells treated with DXR. Intracellular DXR effluxed rapidly from K562/DXR cells, but HSA-ga-DXR as well as HSA-dex-DXR conjugates remained in the cells at a relatively high concentration for a long time. These results indicate that it may be possible to overcome multidrug resistance by chemically modifying DXR, such as by conjugation of the drug with proteins. PMID- 8235309 TI - Serum soluble interleukin-2 receptor assay in epithelial ovarian cancer. AB - Preoperative serum soluble interleukin-2 receptor (sIL-2R) and CA 125 levels were measured in 183 patients with ovarian masses undergoing laparotomy. Serum sIL-2R levels were higher in the 54 patients with epithelial ovarian cancer than in the 129 patients with benign ovarian diseases (p < 0.0001). Elevated serum levels of sIL-2R (> or = 71 U/ml) and CA 125 (> or = 83 U/ml) were found in 79.6 and 77.8% of patients with epithelial ovarian cancer, respectively. Serum sIL-2R and CA 125 positivity rates correlated with the FIGO stage (p = 0.0033 and p = 0.0001, respectively). Raised serum levels of sIL-2R and CA 125 were detected in 11.6 and 7.0% of patients with benign ovarian diseases, respectively. The combination sIL 2R or CA 125 had a sensitivity of 88.9%, and the association sIL-2R and CA 125 had a specificity of 98.4% for epithelial ovarian cancer. As for the 16 patients with this malignancy who were serially monitored during and after chemotherapy, changes in sIL-2R and CA 125 levels correlated with the clinical course of disease in 62.3 and 94.3% of 53 instances, respectively. In conclusion, the serum sIL-2R assay could represent a useful adjunctive tool for the differential diagnosis of ovarian masses, while it seems to be of limited benefit for monitoring the response to chemotherapy and follow-up of patients with epithelial ovarian cancer. PMID- 8235310 TI - Characterization of CA 125 synthesized by the human epithelial amnion WISH cell line. AB - CA 125 has been established as an important tumor marker for monitoring patients diagnosed with nonmucinous ovarian cystadenocarcinoma although it has also been shown to be expressed by other carcinomas, normal epithelial tissues, and fetal tissues. Current evidence implicates a role for CA 125 during early fetal development. The human epithelial amnion WISH cell line is a known secretor of CA 125. WISH cells have been investigated as a model system to characterize the structure of cell-associated and secreted CA 125 of fetal origin. CA 125 secretion was maximal in confluent monolayers of WISH cells where it averaged 2,081 units/ml/24 h. Secretion ranges from 600 to 900 units/10(6) cells/24 h. [35S]-Methionine-labelled CA 125 can be detected by 4 h and reached maximal levels of radioactive incorporation in tissue culture medium by 12 h when analyzed by immunoprecipitation with the M11 anti-CA 125 monoclonal antibody and SDS-PAGE, followed by autoradiography. Both cycloheximide and actinomycin D inhibited CA 125 synthesis. CA 125 was demonstrated to incorporate [3H]-galactose but the level of radioactive incorporation was greatly reduced when WISH cells were incubated in the presence of phenyl N-acetyl-alpha-D-galactosaminide (an inhibitor of O-linked glycosylation) or monensin (an inhibitor of intracellular protein transport within the Golgi complex). Treatment of WISH cells with tunicamycin (an inhibitor of N-linked glycosylation) only slightly decreased label incorporation. PMID- 8235311 TI - Health care reform. Nursing braces for what could be a tough transition. PMID- 8235312 TI - Managing diabetes: a major study injects good news. PMID- 8235313 TI - OTC. GI remedies. PMID- 8235314 TI - New eye into the heart. PMID- 8235315 TI - Did Henry deserve more from us? PMID- 8235316 TI - 1993 earnings survey. Double-digit raises disappear. PMID- 8235318 TI - Here we go again. PMID- 8235317 TI - Staying out of trouble on the telephone. PMID- 8235319 TI - Going to camp: an Rx for burnout. PMID- 8235320 TI - Public awareness can save a profession. PMID- 8235321 TI - What nurses can do to save the environment. PMID- 8235322 TI - Task force seeks RN comments on infection control. PMID- 8235323 TI - A cystic fibrosis patient wants a DNR order. What would you do? PMID- 8235324 TI - Your guide to trouble-free transfusions. PMID- 8235325 TI - Managing AIDS-related meningitis. PMID- 8235326 TI - When gauging respiratory status is critical. PMID- 8235327 TI - Put these on your bookshelf. PMID- 8235328 TI - Emergency cardiac care: the new drug protocols. PMID- 8235329 TI - Organ donation: whose decision is it anyway? PMID- 8235330 TI - AIDS and HIV resources for the illiterate. PMID- 8235331 TI - Join the game. PMID- 8235332 TI - Watch for severe reactions to vitamin K. PMID- 8235333 TI - When you're ready to give up. PMID- 8235334 TI - [Aluminum in drugs]. PMID- 8235335 TI - [Influence of oral administration of aluminum chloride on intestinal transport of leucine and methionine]. PMID- 8235336 TI - [Examining the possibilities of biological utilization of coagulated sludges containing aluminum from water purification plants]. PMID- 8235337 TI - [Tolerance of rye inbred strains on toxic action of aluminum ions]. PMID- 8235338 TI - [Aluminum in infant food]. AB - The potential risk for health because of aluminium content of infant formulas has been taken into consideration. Results of aluminium determination of milk and milk-cereal products and others, from different countries, have been discussed. Primary study on the aluminium content of Polish commercially available infant formulas were performed. The obtained levels were comparable to those reported by other authors. PMID- 8235339 TI - [Toxicology of aluminum]. PMID- 8235340 TI - [Biochemical processes and molecular mechanisms of aluminum toxicity]. PMID- 8235341 TI - [Aluminum in chronic renal failure]. AB - In the paper below are presented the undesirable effects of aluminium in patients with chronic renal failure. Until now aluminum treatment has been used to reduce the level of phosphates in blood serum or in haemodialysis a this group of patients. The toxic activity is a result of cumulation of the element in organism, mainly owing to limited elimination through kidneys, but also as a consequence of disturbed absorption in digestive tract or intravenous infusion of blood serum and albumins. It can be expressed by several clinical syndromes, especially dialysis, encephalopathy, osteomalacia, microcystic++ anaemia, more rarely calcinosis and increased general morbidity and mortality rate. The diagnosis of suspected toxic activity of aluminium in patients with uraemia is based mainly on estimation of concentration of this ion in blood serum and aluminium deposits in skeletal system and skin. The effective treatment consist in intravenous infusions of deferoxamine in a dose 2 g 3 times a week for a long time. PMID- 8235342 TI - [Role of aluminum in the etiopathogenesis of neurological disturbances]. AB - Some more important former and recent views on Aluminium influence on degenerative changes of nervous tissue-including those of Alzheimer's type-were presented. Attention was payed also to Aluminium participation in developing of encephalopathy in dialysed persons and in accumulation of metallic deposits in brain in the so-called Fahr's syndrome. PMID- 8235343 TI - [Aluminum in food]. AB - Literature of the subject has been used to present data on the content of aluminium in groups of food products, and possible sources of its intake. Aluminium content in majority of naturally derived products does not exceed 10 mg/kg (usually 0.1-1 mg/kg). This element is consumed by humans mainly through cereals, cheese and salt. Herbs, spices and tea have a naturally high content of aluminium. Nutrients are a significant course of aluminium in infants and small children. Its content in milk-based mixes is over ten times higher, and in soya based mixes up to several dozen times higher than that reported for breast milk. Aluminium compounds used as food additives are an additional source of this element in food in many countries. Such additives are not permitted in Poland. Food pollution with aluminium may, to some extent, be augmented by use of aluminium cutlery and kitchen utensils, equipment used in food industry, as well as packaging. Consumption analysis presented in 1989 by the FAO/WHO Experts Commission for food additives conclude that the daily intake of aluminium in children is 2-6 mg/kg, and in adults 6-14 mg/kg. The PTWI parameter for aluminium is 7 mg/kg body weight. PMID- 8235344 TI - Aluminium in some surface waters in Poland. AB - This paper describes the results of the study of water acidification and aluminium concentration. High concentration of aluminium was observed in water in Tatra Mts, Karkonosze Mts, Lysogory Mts and on the south side of Barania Mountain. The concentration of aluminium exceeded many times toxic level for water living organisms. PMID- 8235345 TI - Aluminum and metabolism of some plant and animal organisms. AB - Aluminium ions are a serious toxic factor for many organisms living under conditions of the environment. Current views on the mechanisms of aluminium tolerance are presented. PMID- 8235346 TI - [Aluminum as an adjuvant in vaccines and post-vaccine reactions]. AB - Aluminium compounds have been widely used as adjuvants in prophylactic and therapeutic vaccines. Adjuvants are able to stimulate the immune system in a nonspecific manner, i.e. high antibody level can be obtained with minimal dose of the antigen and with reduced number of inoculations. Adjuvants use has been mostly empirically determined by such factors as efficacy and safety. The mechanism of action of the aluminium adjuvants is not completely understood and is very complex. The basic factors of the mode of action: 1) the complex of antigen and aluminium gel is more immunogenic in structure than free antigen, 2) effect "depot"--The antigen stimulus last longer, 3) the production of local granulomas. Vaccines adsorbed onto aluminium salts are a more frequent cause of local post-vaccinal reactions than plain vaccines. 5-10% those vaccinated can develop a nodule lasting several weeks at the injection site. In some rare cases the nodules may become inflammatory and even turn into an aseptic abscess. The nodules persisting more than 6 weeks may indicate development of aluminium hypersensitivity. Finally aluminium adjuvant immunogens induce the production of IgE antibodies. PMID- 8235347 TI - [Therapeutic and toxic properties of pharmaceutical preparations containing Al3+]. PMID- 8235348 TI - [Aluminum in the environment]. PMID- 8235349 TI - [Analytical methods of aluminum determination]. PMID- 8235350 TI - [The effect of aluminum on activity of certain enzymes and on the level of calcium and creatinine in rabbit serum]. PMID- 8235351 TI - [Effects of chronic parenteral administration of aluminum observed in liver of mice]. PMID- 8235352 TI - [Testing the usefulness of using ASV methods for determining aluminum contamination in serum derived products]. PMID- 8235353 TI - [Effect of aluminum on some stages of heme biosynthesis in rats]. PMID- 8235354 TI - [Epidemiology of tobacco smoking in France]. AB - The authors analyse the results of the last national survey on the prevalence of tobacco smoking, carried out by the French Committee for health education in 1992. The smoking percentage is analyzed by sex, age, instruction level and, parents smoking behaviour. These results are also compared with data from previous surveys performed since 1977 in young people and since 1974 in adults. A decreasing trend is noticed in young people from 46% in 1977 to 34% in 1992. This optimistic evolution however has to be relativised, taking into account the increase of the consumption level. For adults, the general trend is the stability of the smoking percentage (40% of the population smoke) but the evolution shows totally different results for males and females. The smoking prevalence is decreasing among men (from 59 to 48%), whereas it is increasing among women (from 28 to 33%). Moreover the smoker percentage among young adult women (18-24 year old) is nearly the same as for young adult men (63.4 vs 63.8%). The data provided by this survey enables the authors to set priorities in the campaign against tobacco smoking. PMID- 8235355 TI - [Toxicology of tobacco]. AB - Tobacco is one of the major toxic agents in our civilization. The extent and severity of the current smoking epidemics are due to the use of industrial cigarettes: their smoke, less irritant than that of cigars and pipe, can be inhaled intensively with rapid absorption of all toxic compounds. Tobacco smoke is composed of a gas phase (CO2, CO, CNH, etc.) and a particle phase (a very fine aerosol) in which more than 4000 substances have been identified. The main toxic compounds in tobacco are Co, nicotine and tars. CO binds to haemoglobin to form COHb, a major factor of hypoxia and vascular accidents. Within a few seconds, nicotine reaches the brain where it binds to specific receptors, which explains its psycho-active effects (psychological dependence) and the induction of physical dependence. This dual dependence is responsible for the failures and relapses observed in attempted withdrawals. Furthermore, nicotine stimulates the sympathetic system with hypersecretion of catecholamines resulting in vascular complications. Tars contain carcinogenic substances, such as aromatic hydrocarbons, which exert local effects on the respiratory tract and systemic effects since they are absorbed by the lungs; this explains the remote neoplasias such as cancer of the bladder. Tars also contain irritant agents (acrolein, formaldehyde, etc.) and oxidative substances responsible for chronic bronchitis and emphysema. Thus, all organs and tissues can be damaged by the toxic compounds that are present in tobacco and, in particular, in tobacco smoke. PMID- 8235356 TI - [Mortality from tobacco use]. AB - The health consequences of tobacco smoking can be estimated from tobacco consumption data, mortality statistics and estimates of risks observed among populations of smokers in cohort studies. In 1990, 60,000 deaths were attributable to smoking in France, corresponding to more than 10% of all deaths. The effect of smoking is much greater in the male (55,000 deaths) than in the female population (5,000 deaths). It is in the age group 45 to 64 that tobacco has the largest effect, contributing to 28% of all deaths in the male population. The health effects observed in 1990 are the consequences of habits which were adopted 20, 30 or 40 years ago. Because the consumption of cigarettes increased regularly between 1950 and 1976, and remained stable since, one can predict that the number of tobacco related deaths will increase for several more decades. This increase will be largest in the female population, and what is already seen in the USA: more women dying from lung cancer than from breast cancer, will one day be seen in France. Despite anti-smoking activities, there is no marked reduction in tobacco sales, and the decrease in the relative price of cigarettes observed between 1960 and 1980, will not be completely compensated by the increases planned for 1993. PMID- 8235357 TI - [Cancers related to tobacco smoking]. AB - Tobacco smoking is the major cause of lung cancer. Cigarette smokers have a risk of lung cancer 10 to 15 times greater than nonsmokers. Tobacco and alcohol are the main risk factors for cancers of oral cavity, larynx, pharynx and oesophagus (cancers of the upper respiratory and upper digestive tract) and the effects of tobacco and alcohol are multiplicative. For these cancers, the risk associated with tobacco was about 2 to 4 among people who drink little or no alcohol. Risks of lung cancer and of cancers of the upper respiratory and upper digestive tract increase with an increasing number of cigarettes smoked per day and duration of smoking. Tobacco is also a risk factor for bladder cancer. Cigarette smoking is a possible contributory factor in the development of kidney, pancreatic and cervical cancers. Among males, lung cancer mortality increased regularly over time and today, lung cancer is the leading cause of death and illness from cancer. Substantial reductions in the number of deaths from tobacco-related cancers could be achieved if a large proportion of smokers stopped smoking. PMID- 8235358 TI - [Tobacco smoking and cardiovascular diseases]. AB - Epidemiological studies have shown that chronic tobacco smoking is associated with a significant increase in risks of coronary disease (chiefly myocardial infarction and sudden death), occlusive arteriopathy of the lower limbs and cerebral vascular accident. The risk is strongly augmented by the presence of other vascular risk factors such as arterial hypertension, hypercholesterolaemia, diabetes and oral contraception. Nicotine and carbon monoxide seem to play a major role in the effect of smoking on vessels. In addition to its acute haemodynamic effects, tobacco not only has an atherogenic effect (endothelial toxicity and changes in lipid profile), but it also facilitates thrombosis (by alteration of platelet functions and elevation of fibrinogen level, haematocrit level and blood viscosity) and spasm (by modification of prostaglandin metabolism and action on catecholamines). All these isolated or associated mechanisms account for clinical and evolutive manifestations or coronary or peripheral arterial lesions. The progression of female smoking partly explains the frequency of vascular pathology in young women. Recent experimental data and epidemiological studies have confirmed the responsibility of passive smoking for coronary pathology, thus justifying the measures enforced to limit the exposure of non-smokers. To cease smoking is probably the most efficient primary or secondary preventive measure, as it results in a relatively rapid decrease in the risk of complications, and notably thrombosis. No effort should be spared to obtain these results and at the same time correct other vascular risk factors. PMID- 8235359 TI - [Chronic respiratory diseases from tobacco use]. AB - Cigarette smoke exerts significant deleterious effects on both lung structures and function. Tobacco alone is the most important risk factor for the development of chronic obstructive lung disease (COLD). The responsibility of smoking for the occurrence of COLD has been demonstrated by epidemiological studies and anatomicopathological findings. Smoking-related lesions initially involve the small airways and remain silent for a long time. There are still uncertainties concerning the mechanisms through which some smokers develop clinical COLD. The clinical diagnosis of COLD rests on the demonstration of dyspnoea by meticulous questioning. Only respiratory function tests can assert the presence of bronchial obstruction. These tests also are necessary to evaluate the severity of the disease, determine its prognosis and follow up its course. In clinical practice the early diagnosis of COLD rests on repeated measurements of FEV1 in smokers. PMID- 8235360 TI - [Passive smoking. Effects on health]. AB - Passive smoking--also called involuntary or environmental smoking--is the exposure of non-smokers to the tobacco smoke released by smokers. The physico chemical composition of tobacco smoke, and notably its contents in toxic and carcinogenic substances, is the same in the secondary stream between puffs as in the primary stream released by the smoker. The pathogenic effects of passive smoking are increasingly well known and accepted. A high incidence of respiratory tract infections and of chronic respiratory and asthmatic symptoms is observed in children. In adults, passive smoking seems to be one of the main risk factors for cardiovascular diseases. Its repercussions on the respiratory tracts is difficult to evaluate, but there are marked by an increase of respiratory symptoms and perhaps of chronic obstructive lung diseases. Finally, it is now recognized that passive smoking is a major risk factor for primary lung cancer in non-smokers exposed to tobacco smoke. PMID- 8235361 TI - [Tobacco and weight]. AB - The weight increase which immediately follows the cessation of smoking is often dissuasive. All things being equal otherwise, the weight and adiposity of smokers are lower than those of non-smokers. The calorie intake of smokers is equal to that of non-smokers, but the nicotine absorbed with each cigarette smoked temporarily increases the basal metabolism and in particular the energy expended in all physical activities. The lack of compensatory increase of food intake in response to the increase of energy expenditure explains why smokers are underweight. Food consumption might be reduced by an increase of circulating fatty acids and by a nicotine-induced inhibition of insulin secretion. The weight increase observed after tobacco withdrawal is caused by the normalization of expenditures and by release of the brake on food consumption. Weight increase is usually important in heavy smokers, women and subjects with low physical activity. Fifty per cent of smokers who have given up smoking put on less than 3 kg. The effort required to mitigate this disadvantage is modest and it can be delayed and separated from the acute weaning period with a medicinal treatment. PMID- 8235362 TI - [Drug aid to tobacco withdrawal]. AB - Persistent tobacco abuse is due to a pharmacological dependence to nicotine and to a psychological dependence to smoking. Tobacco replacement therapies, i.e. nicotine chewing gums and patches, are the only ones that are effective in aiding tobacco withdrawal; they reduce the deprivation syndrome, thereby facilitating smoking cessation and increasing its success rate. The stronger the dependence to nicotine the greater their efficacy. The insufficient results obtained at mid term prove that replacement therapies alleviate only the dependence to nicotine and that the psychological dependence should not be overlooked. PMID- 8235363 TI - [Non-drug aid to tobacco withdrawal]. AB - The smokers who receive the interventions that have often been the source of information in the literature about smoking cessation represent about 4% of the population of smokers: volunteers and help-seekers. This population is not representative of the smokers seen by their doctors; smoking patients can be at any of the stages in their smoking career. But doctors can play an important role in facilitating a smokers's progression towards smoking cessation by a clear, consistent and unequivocable message not to smoke. Depending on the time available and patient desire, doctors can call upon strategies from techniques developed in cessation clinics. The number of contacts with the smoking patient to provide these strategies is the most important factor in the aid the doctor can provide. PMID- 8235364 TI - [Evaluation of smoking prevention programs]. AB - To be generalized, preventive or curative health actions must be efficient. Unfortunately, at this moment in France few promoters of anti-smoking actions have tried to demonstrate the efficiency of the programmes they had proposed. Any evaluation aimed at demonstrating the efficacy of a programme (experimental situation) or its routine applicability requires, like all scientific works, formulation of the objectives, choice of the adequate methodology and list of the tools needed for the analysis. Two levels must be evaluated: the process which includes feasibility, acceptability and efficiency, and the results expressed in terms of changes in behaviour and incidence of target diseases. A few actions which have been evaluated include sensitization by the media, therapeutic assistance in withdrawal, education in schools and legal actions; they are commented in this article. PMID- 8235365 TI - [Biphosphonates in the treatment of osteolytic metastasis. Essential actions on hypercalcemia and important on lytic lesions]. PMID- 8235366 TI - [Respiration through mask. A major progress in the treatment of COBP outbreaks]. PMID- 8235367 TI - [Concomitant radiotherapy and chemotherapy for infiltrating cancers of bladder. Conservative treatment with very promising perspectives]. PMID- 8235368 TI - [Therapeutic perspectives in primary glomerulonephritis from recent physiopathological data]. AB - In this review the authors describe the physiopathological mechanisms responsible for inflammatory lesions in primary glomerulonephritis. Primary glomerular nephropathies can be reproduced experimentally: models have been set up for non proliferative kidney diseases such as minimal change disease or membranous glomerulopathies, and for proliferative kidney diseases such as Berger's disease. Studies performed with these models have resulted in the identification of inflammatory mechanisms creating glomerular lesions which may be acute (proteinuria) or progressive (glomerular sclerosis). These mechanisms include activation of resident cells, notably mesangial cells, and recruitment of circulating cells such as neutrophils, monocytes/macrophages and platelets. These cells participate in the local inflammatory reaction by releasing soluble mediators including biologically active lipids, such as eicosanoids, reactive oxygen and nitrogen derivatives, proteases, cytokines and vasoactive polypeptides. The importance of these mediators has been deduced from the fact that they are produced locally in excess, their introduction produces lesions and, above all, their suppression by inhibitory or antagonistic pharmacological agents reduces the severity of the lesions. Several observations have shown that the development of all the lesions can be prevented simply by blocking the activity of only one of these mediators. It is therefore possible to consider replacing the conventional glucocorticoid treatment, which has multiple pharmacological actions, by a more specific treatment directed against a single mediator. PMID- 8235369 TI - [Neurosis syndrome: obsessive neurosis. Diagnosis, treatment]. PMID- 8235370 TI - [Nitrate derivatives. Principles and rules of use]. PMID- 8235371 TI - [Acute fever (less than 5 days). Diagnostic orientation]. PMID- 8235372 TI - [Infectious meningitis with clear fluid. Epidemiology, etiology, diagnosis, development, prognosis, treatment]. PMID- 8235373 TI - [Acne. Etiology, physiopathology, diagnosis, treatment]. PMID- 8235374 TI - [Alcoholism. Average alcohol content and energy value of wine and beer. Kinetics of alcohol. Epidemiology of consumption and pathology related to excessive alcohol use. Structure of management]. PMID- 8235375 TI - [Non-cirrhotic alcoholic hepatopathy. Diagnosis, development, prognosis, pathological anatomy, treatment]. PMID- 8235376 TI - [Cancer of the esophagus. Epidemiology, etiology, diagnosis]. PMID- 8235377 TI - [Measuring the state of health of the population. Mortality, morbidity, indicators of health, incidence, prevalence, attack levels. Risk, risk factors, risk groups, principal characteristics of the morbidity and mortality in France]. PMID- 8235378 TI - [Erythema nodosum. Diagnostic orientation]. PMID- 8235379 TI - [Erythropoiesis and its regulation]. AB - Erythropoiesis is the process leading to the formation of red cells. Our understanding of erythropoiesis regulation has been due to the characterization of the erythroblast progenitor cells BFU-E and CFU-E. These cells cannot be recognized morphologically, but they proliferate and differentiate in vitro under the effect of growth factors. They correspond to different steps in erythroblast differentiation which is in fact subjected to different regulations. BFU-E cells are the earliest cells of erythropoiesis and are regulated by such growth factors as GM-CSF, Il-3 or Il-9, which are not specific to erythropoiesis. As they differentiate, the BFU-E cells become sensitive to erythropoietin, the erythropoiesis-specific hormone secreted mainly by the kidney in response to hypoxia. The action of erythropoietin and that of other growth factors are synergistically reinforced by a non erythropoiesis-specific growth factor called Steel factor which plays a very important physiological role since defects in its synthesis caused by genetic anomalies produce a lethal anaemia in mice. CFU-E cells, which precede proerythroblasts, are the most sensitive to the action of erythropoietin both in vitro and in vivo. A better knowledge of erythropoiesis regulation mechanisms should help us in the understanding of the physiopathology of some congenital or acquired diseases, such as erythroblastopenia or polycythaemia vera. PMID- 8235380 TI - [Chronic anemia: diagnostic tools and approach]. AB - Anaemia is frequently encountered in daily practice. With full knowledge of its pathophysiology a rational classification is possible allowing a suitable approach for diagnostic investigations. In a first stage, the data provided by blood counts, erythrocyte constants and reticulocyte counts guide the diagnostic rationale. In cases with microcytic and hypochromic anaemia, measurement of ferritin level separates iron deficiency anaemia from the so-called "inflammatory" anaemias. A high number of reticulocytes points to haemolytic anaemia. Among the many causes of haemolysis, one must first look for autoimmune haemolysis. Elsewhere, the clinical context and morphological red cell abnormalities point to a hereditary disease affecting the erythrocyte membrane, enzymes or globin content. Although rare, microangiopathic anemia with schizocytosis and paroxysmal nocturnal haemoglobinuria must not be misdiagnosed. Bone marrow examination remains the clue in non-regenerative normochromic, normo- or macrocytic anaemias. In difficult cases, other investigations, such as cytogenetics, isotopic examination and progenitor culture, may help in characterizing the anaemia. Serum erythroproietin essays and plasma transferrin receptor counts will soon figure among the methods used to explore anaemias. PMID- 8235381 TI - [Inflammatory hyposideremic anemia]. AB - Inflammatory low iron is the second cause, after true iron deficiency, of acquired anaemia. It is mainly due to insufficient erythropoiesis resulting from inhibition of the erythroid progenitor and to disturbances in the synthesis and action of erythropoietin. These changes seem to be dependent on factors, such as TNF-alpha, interleukin-1 and interferon-gamma, which are released in inflammatory processes. Alterations in iron metabolism seem to be secondary, but also partly provoked by the same inhibitory agents. All these anaemias share a common character, i.e. lowering of serum iron level without increase of transferrin level, while plasma ferritin level is within normal limits. In addition to symptomatic therapy by red cell transfusions, numerous trials have shown that recombinant erythropoietin is effective in the treatment of the anaemia that accompanies cancers, chronic inflammatory and rheumatic diseases and of the anaemia provoked by HIV infection. PMID- 8235382 TI - [Iron deficiency anemia]. AB - Iron deficiency anaemia is characterized by the conjunction of a microcytic, typically are generative anaemia and a biochemical syndrome in which sideropoenia is associated with transferrin increase. It is usually due to a chronic exsudative gastrointestinal or gynaecological bleeding. Diagnosing the mechanism of anaemia is easy in most cases, but difficulties arise from association with a pathology, such as chronic inflammatory syndrome, disturbing the haematological or biochemical data. The prognosis of iron deficiency anaemia depends on its cause. Treatment is aetiological (the cause of chronic bleeding is suppressed) and symptomatic (the body's iron reserves are quickly restored). PMID- 8235383 TI - [Anemias due to disorder of folate, vitamin B12 and transcobalamin metabolism]. AB - Macrocytic megaloblastic anemia is the most typical but the latest sign of a cobalamin (vitamin B12) and/or folic acid deficiency or of a congenital abnormality of cobalamin and folate metabolism. Macrocytosis in blood and megaloblastosis in bone marrow are the morphological features of a disturbance in cell division related to a defect in DNA biosynthesis. Macrocytosis without anemia, normocytic normochronic anemia with a low reticulocyte cell count or microcytic hypochromic anemia in case of associated iron deficiency do not exclude a vitamin deficiency. Neurological or psychiatric disorders and immune abnormalities have been reported in patients with vitamin B12 or folate deficiencies or in children with congenital abnormalities of these 2 vitamins; such manifestations may even occur without anemia. PMID- 8235384 TI - [Immune hemolytic anemias (autoimmune, drug-induced)]. AB - The serological investigation of immunohaemolytic anaemias is based on the direct antiglobulin test, on the study of the serum and on the eluate, thus allowing a classification, which is still valid and correlates well with the clinical features. The use of more sensitive techniques and of reagents made from monoclonal antibodies should improve the study of these anaemias, but emphasizes the issue of the limits between physiological and pathological states. The addition of new therapeutical approaches should improve their prognosis. The increasing consumption of drugs and the development of the investigations of the adverse reactions including the haemolytic anaemias have allowed the emergence of new pathogenic concepts. The severity of the course of some of these immune haemolytic anaemia stresses the importance of an adequate therapeutic survey and the development of new means of diagnostic. The clinicians should be aware of the possibility of cross reactivity for drug dependent antibodies with other components of related chimical structure. PMID- 8235385 TI - [Anemias due to erythroblastopenia]. AB - Anaemias caused by pure red cell aplasia may regress spontaneously (acute) or remain chronic. They have multiple causes, the most frequent of which, in chronic pure red cell aplasia, is chronic lymphocytic leukaemia. It is important to find that the anaemia is due to pure red cell aplasia since this may raise hopes of cure after an immunosuppressive treatment. In patients with chronic pure red cell aplasia the in vitro study of erythroid precursors has a prognostic value. PMID- 8235386 TI - [Paroxysmal nocturnal hemoglobinuria]. AB - Paroxystic nocturnal hemoglobinuria (PNH) is an acquired hemolytic anaemia related to an increase susceptibility of erythrocytes to complement-mediated lysis. PNH is a clonal disease of an hematopoietic stem cell which lost, by mutation, the ability to synthesized phospholipid anchor of membranous proteins, i.e. complement regulatory proteins: DAF, C8BP or CD59. The clinical features of PNH are hemoglobinuria episodes associated with chronic hemolytic anaemia or pancytopenia with active bone marrow or aplastic anaemia. The clinical course is marked by severe thrombotic complications (such as Budd-Chiari syndrome), hemorrhages or infections. The diagnosis is confirmed by in vitro hemolysis tests, and now by facs analysis of cell membrane expression of deficient proteins. Different treatments have been proposed with various results (corticosteroid therapy, androgens, chemotherapy...) but the only way to eliminate the abnormal clone appears to be related bone marrow allograft. PMID- 8235387 TI - [Myelodysplastic syndromes or refractory anemias]. AB - Myelodysplastic syndromes are relatively frequent, with a distinct predominance in elderly subjects. They are characterized by a disorder of myeloid precursor cell maturation, which explains the presence of blood cytopenia responsible for clinical manifestations (anaemia, infection, haemorrhage). Beside cytopenia, the main risk is transformation into acute myeloid leukaemia. As a rule, these diseases are easily recognized by the conjunction of blood count and bone marrow aspirate. Apart from the intensive therapy prescribed for myelodysplastic syndromes in young subjects, treatments seldom have beneficial effects and are still symptomatic in most cases. PMID- 8235388 TI - [Myelodysplastic syndromes in children]. AB - Myelodysplastic syndromes are clonal haemopathies known to exist in elderly people where they are classified into 5 categories: refractory anaemia, acquired idiopathic sideroblastic anaemia, refractory anaemia with excess of blasts, refractory anaemia with excess of blasts undergoing acute transformation, and chronic myelomonocytic leukaemia. Transformation into acute leukaemia is frequent. These syndromes seem to be rarer in children, but they are often misdiagnosed. Some of their forms are particular to childhood. They include association with constitutional blood diseases predisposing to acute leukaemia, or with congenital or non-congenital malformative syndromes; there are hypoplastic forms, forms evolving towards stabilization of haematological abnormalities, and border forms with certain myeloproliferative syndromes. The diagnosis of myelodysplasia rests on morphological, functional, biochemical, isotopic data reflecting inefficient haematopoiesis, and on cytogenetic data. Studies of clonality and malignancy markers will in the near future enable us to distinguish between true preleukaemic states and polyclonal constitutional or virus-induced myelodysplasias, probably more frequent in children. The choice of treatment depends on the severity of the myelodysplastic syndrome, defined by the initial partial blastosis, the presence of cytogenetic anomaly and the certainty of clonality. In cases with poor prognosis, early bone marrow allograft is the only possible treatment. PMID- 8235389 TI - [Molecular pathology of the erythrocyte membrane]. AB - The erythrocyte membrane, in the broad sense of the term, comprises a lipid bilayer and below it the erythrocyte skeleton responsible for the mechanical properties of red cells. The lipid bilayer contains numerous proteins including the anion transporter. The underlying portion contains spectrin, actin and protein 4.1. At the interface between these two portions, ankyrin, protein 4.2 and other proteins are found. The structure and mode of expression of the corresponding genes are roughly known, as are the structure-function relationships within the proteins. In the wake, or sometimes ahead of these fundamental advances, dramatic strides have been made in the molecular pathology of the erythrocyte membrane. Hereditary spherocytosis results from mutations altering on ankyrin, the anion transporter or protein 4.2, while elliptocytosis results from mutations affecting spectrin or protein 4.1. Thus, the knowledge already acquired makes it possible to diagnose, at molecular scale, not only hereditary spherocytosis and elliptocytosis, but also several other genetic diseases of the erythrocyte membrane. PMID- 8235390 TI - [Hemolytic anemias due to enzymopathies]. AB - The most inherited enzymopathies associated with hemolytic anaemias are due to enzyme deficiencies (as a consequence of either synthesis reduction or impairement of the enzyme function). Such deficiencies can disturb some metabolism of the red cells. The most of enzyme deficiencies are involved in glycolysis that represents the main source of energy for this cell. Generally the genetic transmission of these enzymopathies is recessive autosomal with the exception of two cases (glucose 6-phosphate dehydrogenase and phosphoglycerate kinase) which are X-chromosome-linked. It stands to reason that the only treatment of these enzymopathies is dependent on gene therapy, a fine future method. Nevertheless prenatal diagnosis can be considered in some cases that display particularly severe symptoms. PMID- 8235391 TI - [Blood transfusion in anemic patients. When and how?]. AB - The functional red cells infused must breathe and circulate. The effectiveness of transfusion depends on different factors in patients (lack of antibodies or hypersplenism) and on the way the blood has been preserved. The transfusion related risks may be immediate (immunological conflicts between erythrocytes or leucocytes) or delayed (alloimmunization induced by transfusion, transmission of or viral or parasitic diseases). The current preventive measures are highly efficient, provided all "safety catches" are respected, including selection of donors on immunological and serological grounds, and choice of the red cell concentrate most appropriate to the clinical situation and the transfusional future of the patient. All patients on chronic transfusion must be under long term surveillance, with search for antibodies and serological profile. The decision to transfuse blood rests on the levels of erythrocyte constants and on clinical tolerance to anaemia, taking into account the compensatory mechanisms. Several formulas are available to calculate the red cell volume to be transfused in relation to haemoglobin values. The advances achieved in blood perfusion are such that anaemic patients nowadays are both less and better transfused. PMID- 8235392 TI - [Immunoscintigraphy. Functional imaging for the future]. PMID- 8235393 TI - [Some acquired facts apropos of the effectiveness of heparins in the prevention of postoperative phlebitis and pulmonary embolism]. PMID- 8235394 TI - [Imaging of scoliosis]. AB - The majority of scoliosis (80%) and especially the idiopathic scoliosis are simply investigated by plain films of the entire spine with erect frontal and lateral views, and supine frontal view at the initial examination. Repeated erect frontal views are sufficient for the follow-up. The important radiation dose (average of 1.2 mG for the frontal view and 2.2 mG for the lateral views) imposes to reduce the frequency of these radiographs. This radiological study allows: to separate non structural and structural curves; the identification and the measurement of a structural curve, discussion for its etiology, appraisal of its progressiveness, discussion of the operative or non operative treatment, and radiological follow-up. Modern imaging (C.T, C.T myelography, MRI) is indicated only in very particular cases: idiopathic scoliosis with neurological symptoms, congenital scoliosis with structural vertebral abnormalities, especially these with posterior arch abnormalities, and progressive scoliosis in spinal dysplasiae. Painful scoliosis are unusual. If pain is present consideration should be given for diagnosis of an underlying lesion. In this case a relevant strategy using modern imaging becomes useful and will be discussed. PMID- 8235395 TI - [Bedridden state. Etiology, prevention, management]. PMID- 8235396 TI - [Pruritus (except anal and vulvar). Diagnostic orientation]. PMID- 8235397 TI - [Tetanus. Physiopathology, diagnosis, prevention]. PMID- 8235398 TI - [Complications of biliary calculi. Etiology, diagnosis, treatment]. PMID- 8235399 TI - [Gastro-esophageal reflux. Physiopathology, diagnosis, development, treatment]. PMID- 8235400 TI - [Headache. Diagnostic orientation]. PMID- 8235401 TI - [Acute pancreatitis. Etiology, diagnosis, development]. PMID- 8235402 TI - [Atrial activity and its effects]. AB - The initiation of cardiac impulse is located in the sinus node, in the upper anterior part of the right atrium. The importance of the atrium is not only linked to the regulation of heart rate, but also to its haemodynamic function. Indeed, atrial depolarization leads to atrial contraction which can be responsible for up to 30% of cardiac output by way of ventricular filling. Supraventricular arrhythmias are related to one of the following mechanisms: abnormal automaticity, triggered activity, and reentry. Most of supraventricular tachycardias are due to a reentrant phenomenon (intranodal reentrant tachycardia, orthodromic circusmovement tachycardia, atrial flutter and atrial fibrillation). At the onset of a supraventricular tachycardia, the loss of efficacious atrial contraction as well as the increased heart rate may abruptly decrease ventricular filling. As a consequence, stroke volume is reduced, leading to a decrease in cardiac output and in arterial blood pressure, explaining that the patient may experience syncope. Usually, blood pressure reduction resumes within 30 seconds after activation of the autonomic adrenergic nervous system. In case of an underlying heart disease, the supraventricular tachycardia may lead to acute cardiac failure. When reentry is concerned, the tachycardia is going around a specific circuit. The existence of such a circuit in most of supraventricular tachycardias has led to the development of ablation therapy, the goal of which is to destroy a critical portion of the circuit hence making the recurrence of reentrant tachycardia impossible. PMID- 8235403 TI - [Non-invasive exploration methods of supraventricular arrhythmia in current practice]. AB - In patients with supraventricular rhythm disorders ambulatory electro cardiographic recording (Holter system) is an indispensable examination as it detects attacks that pass unrecorded by conventional ECG, being asymptomatic, too brief or too rare. It confirms the diagnosis, defines the factors triggering the attacks, detects the association of rhythm and conduction disorders, guides the treatment and monitors its effectiveness. Sequential ambulatory recording lends itself particularly well to this last objective. Biochemical examinations explain the cause of certain relapses (potassium depletion, high alcohol blood level) or detect the origin, clinically more or less obvious, of these disorders of rhythm (essays of thyroid hormones). Measuring blood levels of therapeutic drugs makes the handling of these various drugs safer. Finally, echocardiography detects an underlying heart disease, evaluates the size of the left atrium (a factor of relapse when it is dilated and of embolism when it harbours thrombi) and assesses the left ventricular function before administration of antiarrhythmics which, to varying extents, are all negative inotropic drugs. PMID- 8235404 TI - [Electrophysiological diagnosis of supraventricular tachycardia]. AB - Supraventricular tachycardias are frequent but seldom life-threatening. The role played by electrophysiological exploration in the evaluation of patients is still restricted. The exploration can be performed by the intracavitary or transoesophageal route. The problem is radically different in patients with d atrial arrhythmias or junctional tachycardias, where this exploration aims at determining the degree of atrial vulnerability in the first instance and the exact mechanism of tachycardia in the second. Electrophysiology is indicated principally in junctional tachycardia recurring despite medicinal treatment, when radical ablative therapy is contemplated. In atrial arrhythmias, it is used to study the arrhythmogenic substrate in the forms that resist empirically prescribed treatments and also in young subjects with embolic cerebrovascular accident of unknown origin. PMID- 8235405 TI - [Attacks of junctional tachycardia: from arrest of crisis to radical cure]. AB - Junctional tachycardias are among the most frequent or paroxysmal supraventricular tachycardias. They are due to a reentry mechanism and include the so-called nodal reentrant tachycardias as well as tachycardias which imply a patent or hidden accessory pathway. The prognosis of these tachycardias is usually benign, but it can be made unfavourable by repeated attacks or by the presence of an accessory pathway with short anterograde refractory period which exposes the patient to severe arrhythmia. Clinicians are now provided with a therapeutic armentarium that enables them to reduce easily any attack of junctional tachycardia, but also to prevent recurrences. The intracavitary ablation technique by application of radiofrequency currents ensures the radical cure of recurrent or threatening arrhythmias by suppressing the indispensable anatomical substrate of tachycardias. PMID- 8235406 TI - [Atrial flutter and tachy-systole]. AB - Atrial flutter and tachysystoles may complicate the course of all heart diseases or may be discovered independently of any detectable cardiopathy. Functional tolerance mainly depends on the presence and severity of the associated heart disease, if any. A regular cardiac rhythm of 150 beats/minutes at rest suggests a 2:1 block flutter. Electrocardiographic identification of the disorder is usually easy and may lead, in certain cases, to vagal manoeuvres to recognize the atriogramme and the atrium-ventricle relationship. In all cases, 2:1 flutters must be reduced with amiodarone, by electrical stimulation or by electric shock. A prophylactic treatment of recurrences is often, but not always, indicated. The treatment of tachysystoles must take into account the cause of the ventricular rate. PMID- 8235407 TI - [Paroxysmal atrial fibrillation]. AB - Paroxysmal atrial fibrillation is the most frequent of cardiac arrhythmias and its frequency increases with age. Its diagnosis is difficult since the symptoms are often absent or misleading. It is based on electrocardiogram performed during the attack, and this has been made easier by the Holter technique or by sequential recordings triggered by the patient himself. It is only in special cases, when percritical recordings could not be made that intracavitary electrophysiological study is used to obtain indirect arguments in favour of the diagnosis. All heart diseases can be attended by this cardiac rhythm disorder which usually marks an unfavourable turn in the course of the disease, with an increased risk of sudden death in the end. But in most cases the only disorder found is a common arterial hypertension. Atrial fibrillation carries a risk of peripheral, and in particular cerebral, embolism evaluated at 5% per annum. This risk is high in patients with cardiopathy, but it also exists even with a normal heart. The value of anticoagulants has been demonstrated by large controlled studies, but it is important to know that a very strong anticoagulant treatment is unnecessary to reduce the risk of embolism. Only antiarrhythmic agents can maintain the patients in sinus rhythm for as long as possible before the passage to permanent fibrillation. Although no beneficial effect on mortality has been proved, these drugs are indicated for patients who have very marked symptoms or a major cardiopathy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235408 TI - [Antiarrhythmic treatments of permanent atrial fibrillation]. AB - The treatment of chronic atrial fibrillation has 3 different objectives: the control of the ventricular rate, the attempts to restore sinus rhythm, and the prevention of arrhythmia recurrences. Digitalis compounds, beta-adrenergic antagonists and calcium-channel blocking agents can be used to achieve reduction of the ventricular response, in recent-onset arrhythmia with rapid heart rate, and for long-term rate control in patients who cannot be converted to sinus rhythm. In some of them, a combination of 2 of these drugs is needed to slow the ventricular rate, at rest and on exertion. Sinus rhythm can be restored by direct current cardioversion or by using various regimens of amiodarone, a type III antiarrhythmic drug, given orally or intravenously. Cardioversion might also be accomplished by type IA and IC antiarrhythmic drugs, including quinidine, flecainide and propafenone. However, proarrhythmic effects and haemodynamic deterioration are a major concern with these compounds. On the other hand the best indication of type IA and IC antiarrhythmic drugs is the control of arrhythmia recurrences; beta-blocking agents are highly effective in patients with a high sympathetic tone or hypertrophic cardiomyopathy. Finally the widespread use of amiodarone should be limited by its non-cardiac toxicity. Whatever the drug employed in the treatment of atrial fibrillation, monitoring the inotropic effect of the drug and the effect on cardiac conduction is mandatory. PMID- 8235409 TI - [Antithrombotic treatment of permanent atrial fibrillation]. AB - Whether idiopathic or associated with an underlying heart disease, complete arrhythmia induced by atrial fibrillation ranks first among the causes of arterial embolism. It has been recognized that 30 to 35% of subjects with arrhythmia develop a transient or permanent ischaemic cerebral accident, and it is therefore rational to assess the value of systematic antithrombotic treatment in the prevention of such accidents. It is agreed by all that anticoagulants are necessary when atrial fibrillation occurs in the course of cardiac valve diseases, notably those of rheumatismal origin. Conversely, the idiopathic complete arrhythmia, which usually occurs in subjects aged below 60, calls for either therapeutic abstention or prescription of antiplatelet drugs. In all other cases, five randomized trials can be used as reference. They all show that thromboembolic accidents and mortality can be reduced and favour a systematic preventive treatment. The risk of haemorrhage has been diversely evaluated in these five trials. The choice between anticoagulant and antiplatelet therapies therefore rests on a case by case analysis aimed at balancing in each patient the benefits expected against the possible risks. PMID- 8235410 TI - [Disorders of supraventricular rhythm in children]. AB - In children with otherwise normal heart, reentry is the most common cause of supraventricular tachycardia. The tachycardia is stopped by vagal stimulation, and digoxin is then given to prevent recurrences; in patients with Wolff Parkinson-White syndrome, transeosophageal stimulation is performed to examine properties of anterograde condution across the accessory connection, and to select the most appropriate treatment. Radiofrequency ablation is selected only in children with refractory supraventricular tachycardia or who were admitted for syncope due to rapid conduction over an accessory pathway. Neonatal junctional ectopic tachycardia may be very severe and even results is death when the heart rate is not slowed down; amiodarone is the safest and the most effective drug. Atrial flutter is encountered in 2 groups of children: neonates, in whom it is converted by rapid transeosophageal pacing, with excellent prognosis, and children who have had atrial surgery (Mustard and Senning operations). These patients have sinus failure, with paroxysmal atrial flutter possibly responsible for sudden death when rapidly conducted to the ventricles. It is a new and concerning pathology, as these children have chronic arrhythmias requiring aggressive therapy such as amiodarone and cardiac stimulation. PMID- 8235411 TI - [Dislodgement of articular prosthesis. Role of mediators of inflammation]. PMID- 8235412 TI - [Mutants of hepatitis B virus. Description and value of a better understanding of this disease]. PMID- 8235413 TI - [Use of pancreatic extracts in chronic pancreatitis]. PMID- 8235414 TI - [Arterial hypertension and hypokalemia]. AB - The occurrence of hypokalemia in association with high blood pressure is suggestive of primary hypermineralocorticism since in this case both abnormalities might result from a single mechanism. However, adrenal adenomas is well as the various forms of adrenal hyperplasia appear to be quite uncommon, whereas a number of other causes of potassium depletion are far more prevalent and may be associated with essential hypertension. The demonstration of the precise mechanism of both decreased serum potassium and increased blood pressure is a prerequisite for a successful treatment. PMID- 8235415 TI - [Epidemiology of contagious diseases. Methods of surveillance, notification, investigation of outbreak in a community and preventive measures]. PMID- 8235416 TI - [Rhizomelic pseudo-polyarthritis. Diagnosis and treatment. Horton's disease. Pathological anatomy, diagnosis, development, treatment]. PMID- 8235417 TI - [Pulmonary embolism. Etiology, physiopathology, diagnosis, clinical course and treatment]. PMID- 8235418 TI - [Alcoholism: acute and chronic intoxication, withdrawal syndrome. Epidemiology, diagnosis, principles of treatment (including prevention)]. PMID- 8235419 TI - [Periosteal or juxtacortical chondroma of the mandibular condyle]. AB - One case of juxtacortical chondroma is reported, and the authors review the clinical, radiological, pathological and therapeutic data concerning this tumour which was first individualised by Lichtenstein and Hall in 1952 but never described in this location. Conservative surgery with complete excision of the chondroma usually results in permanent cure. PMID- 8235420 TI - [Central odontogenic fibroma]. AB - Odontogenic fibroma is an infrequent benign tumor, originating in the mesenchyma and including an inactive odontogenic epithelial component. The central form, of which one case is reported here, is expressed by a relatively well-delineated radiolucent area, whose interest and difficulty lie in the differential diagnosis. On the basis of a review of the literature, details are given on the clinical, clinicopathologic, diagnostic and therapeutic data. PMID- 8235421 TI - [A brown tumor of the mandible and hyperparathyroidism secondary to chronic renal insufficiency. Review of the literature apropos of a case]. AB - We report on a brown tumor of the mandible in a hemodialysed female patient with severe uncontrolled secondary hyperparathyroidism. Multiple other brown tumors were associated. The hyperparathyroidism was treated with a subtotal parathyroidectomy because only three parathyroids were found. Parathyroid removal led to normal C-terminal parathyroid hormone concentration (0 ng/ml). A review of the literature has shown that parathyroidectomy is usually curative. Our patient demonstrated, eleven days after the parathyroid surgery, rapid growth of the mandibular tumor. This unusual evolution has justified a local excision of the tumor. Histological preparations have confirmed the diagnostic of brown tumor with an important capillary proliferation. PMID- 8235422 TI - [Toxic epidermal necrolysis associated with lymphoproliferative syndromes. Anatomo-clinical study. Presentation of 2 cases]. AB - Two cases of Toxic Epidermal Necrolysis, with a previous history of drugs intake, and associated to a lymphoma and to a chronic lymphoid leukemia, are presented. The oral lesions are described and the clinical and pathological particularities of the cases are commented. The rule of the drugs and of the neoplastic diseases in the aetiology of these cases is discussed. In conclusion, we emphasize the N.E.T. as an entity of stomatological interest. PMID- 8235423 TI - [Lip ulceration in paraneoplastic dermatomyositis. Apropos of a case. Review of the literature]. AB - A 71 year old woman with metastatic ovarian cancer developed ulceration of the lips during the course of a paraneoplastic dermatomyositis. The principal diagnostic features of multiple myositis are described, as well as the relationship of buccal lesions to the dermatomyositis. Data are lacking on the frequency of these polymorphous and non specific lesions. PMID- 8235424 TI - [Saethre-Chotzen syndrome. A study apropos of 17 cases]. AB - Case reports of seventeen patients with the Saethre-Chotzen syndrome treated at the Stomatology and Maxillofacial Surgery Clinic, Nantes, France are analyzed, together with clinical and paraclinical signs of this syndrome, included within the framework of the acrocephalosyndactylies. Results of cases reported in the published literature are reviewed and compared with those of the present series. The characteristic craniofacial equilibrium of the Saethre-Chotzen syndrome is discussed, together with findings of a teleradiographic study performed according to Delaire's analysis. A discussion on the differential diagnosis of this syndrome is enlarge to include all acrocephalosyndactylies. PMID- 8235425 TI - [Bone age and maxillo-mandibular dysharmony in children with cleft lip and palate]. AB - The clinical examination, the bone age and the palatine surgical trauma served to evaluate the factors producing maxillo-mandibular dysharmonies in cleft lip/palate patients. 55 patients with total unilateral cleft lip and palate aged from 10 months to 13 years were used for this study. By clinical examination the patients were divided in: group A--37 cases without dysharmonies and B--18 cases with dysharmonies. By the bone age examination: group N--39 cases with normal variations (+/- 1.99 SDS) or acceleration (+2.00 SDS) and group R--16 cases with retardation (-2.00 SDS). By the surgical trauma: group M--11 cases with multiples operations and K--44 cases as "control" group. The correlations between the three factors showed that for the prognosis of maxillo-mandibular dysharmonies are significant: 1. The surgical trauma--73% of the patients with multiples operations (group M) are also part of the group B (with dysharmonies), versus 23% for the "control" group K (p < 0.001). 2. The retardation of the bone age--50% of the patients of the B group have retarded bone age, versus 24% of the patients of the group A (p < 0.05). 56% of the patients of the group R (with retardation) are part of the group B, versus 23% of the group N (p < 0.02). The retardation of the bone age is a factor of risk for maxillo-mandibular dysharmonies in cleft lip/palate children, which means that in these cases, the treatment and the control should be more attentive, beginning as soon as possible. PMID- 8235426 TI - [Complex reconstruction of the lower lip. Apropos of various cases]. AB - Preservation of buccal cavity relations raises problems when treating extensive defects of lower lip. After a review of anatomical features, several case reports are presented to illustrate the advantages and inconveniences of three interventions for labial reconstruction. PMID- 8235427 TI - [An auricular prosthesis attached by intra-mastoid implants. Apropos of 11 cases]. AB - The use of osseointegrated implants to provide support for auricle prosthesis as been carried out by Tjellstrom. This method has provided the maxillofacial surgeon with another approach to the treatment of defect of the auricle. The principles and concepts of extra-oral osseointegrated implants are the same as those of the intra-oral application developed by Branemark 26 years ago. This simple and reliable method allow the attachment to splint bar fixated to three or two mastoid osseointegrated implants. This method represent an interesting possibility for the management of auricle defects. We report on our experience of 11 cases. PMID- 8235428 TI - [External rhinoplasty. 250 cases over a 6-year period]. AB - Description by the authors of their experience on external rhinoplasty in 250 cases, the advantage of this technique, compared to the classical extramucosa rhinoplasty and its main indications. PMID- 8235429 TI - The Gastroenterology Prize for 1993. PMID- 8235430 TI - The duodenojejunal junction and Treitz ligament in the regulation of duodenal emptying. AB - In chronic experiments in dogs the evacuatory and absorptive duodenal functions and the level of duodenogastric reflux (DGR) in sham-operated dogs and dogs with transected Treitz ligament (TL) and resected duodenojejunal junction (DJJ) were studied. The electrophysiologic characteristics of the antrum, duodenum, and DJJ in sham-operated dogs were examined. It was shown that there is a coordination of gastric and duodenal solid carbohydrate food emptying. The process of evacuation from both organs was characterized by three closely correlated phases, phase 2 of emptying being exponential. In the TL transection and DJJ resection groups the coordination of gastric and duodenal emptying disappeared. The rate of carbohydrate solid food evacuation from the duodenum decreased, on average, by 45.5% +/- 5.5% (P < 0.05); phase 2 of duodenal emptying became unexponential, and the level of DGR considerably increased. The rate of duodenal absorption of main organic chyme components increased significantly in most of the time intervals studied. Significantly higher amplitudes of slow waves and spike potentials were recorded from the DJJ than from the duodenum. We conclude that the DJJ and TL are possible regulators of duodenal emptying. The DJJ is probably a functional sphincter that provides coordinated activity of the stomach and duodenum. PMID- 8235431 TI - Gastric mucosal blood flow and neutrophil activation in aspirin-induced gastric mucosal damage in man. AB - Gastric and intestinal injury induced by nonsteroidal anti-inflammatory agents (NSAIDs) such as aspirin (ASA) is a common side effect of this class of drugs, but the mechanism by which these drugs act is not fully explained. In this study the effects of 3 days of continuous oral ASA administration (1 g twice daily) to eight healthy male volunteers were studied. To estimate the extent of mucosal damage, gastroscopy was performed before and after 3 days of ASA treatment, during which the mucosal blood flow was measured by means of laser-Doppler flowmetry. Before each endoscopy gastric microbleeding was measured. Since neutrophil activation has recently been suggested to be involved in the pathogenesis of ASA-induced gastric mucosal damage, we examined the influence of ASA treatment on the activation of leukocytes by determining their association with platelets in the blood. Aspirin-induced acute gastric damage reached about 3.5 in the endoscopic Lanza score. Mucosal blood flow increased significantly after ASA treatment, by about 50% in the oxyntic gland area and by 87% in the antral area. Gastric microbleeding rose from about 0.38 ml/day in the intact stomach to about 7.7 ml/day after ASA treatment. The platelet/neutrophil adherence increased significantly in both thrombin-unstimulated and thrombin stimulated platelets. We conclude that acute 3 days' administration of ASA in man produces well-defined areas of gastric damage accompanied by a significant increase in gastric microbleeding and gastric blood flow and that ASA promotes platelet/neutrophil adhesion that may resemble the neutrophil/endothelium interaction in the gastric mucosa. PMID- 8235432 TI - Circadian acidity pattern in prepyloric ulcers: a comparison with normal subjects and duodenal ulcer patients. AB - We used continuous 24-h pH monitoring to compare the circadian intragastric acidity of 36 patients with prepyloric ulcers (PPU) with that of 101 normal subjects (NS) and that of 206 patients with duodenal ulcer (DU). The ulcer crater was endoscopically ascertained in all cases, and PPU were located within an area up to 2 cm proximal to the pylorus. The pH curve pertaining to DU patients ran below that of NS during most of the circadian period, whereas the pH profile of PPU patients was higher than that of NS, and this was particularly true during the evening and the night. The acidity of PPU patients was significantly lower (p < 0.01) than that of NS during the night only, whereas it was lower (p < 0.05 0.001) than that of DU patients during each time interval analysed (24 h, nighttime, and daytime). Our findings show that the gastric acidity of PPU patients differs greatly from that of DU patients, since it is lower throughout the whole 24-h period, and particularly during the night. Thus these two entities are pathophysiologically different with regard to the acidity pattern and should be considered two distinct subgroups of peptic ulcer disease instead of being incorporated, as usually happens, in the clinical group 'duodenal ulcer disease'. PMID- 8235433 TI - Scintigraphic study of gastrointestinal transit and disintegration sites of mesalazine tablets labeled with technetium-99m. AB - Tablets of mesalazine covered with a pH-dependent coating (Pentacol), labeled by an original technique with technetium-99m, were administered to 12 patients, 9 with Crohn's disease, 3 of which recurrent, 1 with ulcerative colitis, and 2 with irritable bowel syndrome, with the aim of verifying in vivo the intestinal site of disintegration and how the contents spread throughout the intestine. In all cases the tablet was broken down in the distal ileum at extremely variable intervals, from 5 to 27 h, and the contents spread into the nearby loops and into the colon. The notable differences in the residence time of the whole tablet in the ileum can be explained by differences in adhesion to the inflamed mucosa and by a lower pH in the part of the ileum affected by the disease. PMID- 8235434 TI - Postprandial conjugated and unconjugated serum bile acid levels after proctocolectomy with ileal pouch-anal anastomosis. AB - In patients with ileal pouch-anal anastomosis (IPAA) bile acid reabsorption may be impaired, and stasis may lead to deconjugation and dehydroxylation of bile acids as a result of bacterial overgrowth. We therefore studied fasting and postprandial conjugated and unconjugated serum levels of cholic (CA), chenodeoxycholic (CDCA), and deoxycholic acid (DCA) in 11 patients who underwent proctocolectomy with IPAA and in 11 healthy controls. Fasting levels of conjugated DCA but not CA and CDCA were significantly lower in IPAA patients. Postprandially, conjugated bile acid levels were significantly lower in IPAA patients. Postprandial unconjugated CA levels were significantly higher and CDCA levels tended to be higher in IPAA patients, whereas unconjugated DCA levels were lower in IPAA patients. These data suggest that reabsorption of conjugated bile acids is impaired after IPAA; deconjugation of bile acids may result from bacterial overgrowth secondary to stasis in the pouch; and dehydroxylation of bile acids is decreased after proctocolectomy with IPAA. PMID- 8235435 TI - Intra- and peri-hepatic effects of ethanol injections in the pig liver. AB - We studied the intrahepatic (spread and scarring) and perihepatic effects of percutaneous ethanol injection therapy and its dose-necrosis relationship in normal pig livers. Ethanol (99.5%) was injected in volumes of 0.5-2.0 ml into the livers of six pigs during laparotomy. The livers were dissected and studied histopathologically 30 min and 2 weeks after the injections. Ethanol-induced liver necrosis, the diameter of which could roughly be calculated by dividing the injected dose by 2 (inversely: volume (ml) = 2 x diameter (cm)). Two weeks after the injections all pigs showed intra-abdominal adhesions and severe intrahepatic scar formation, unlike three control pigs similarly operated on without ethanol injections. The 30-min lesions showed thin projections extending into the neighboring liver tissue. Major escape of ethanol outside the needle tip area in the liver occurred twice. Uncontrolled spread of ethanol may be a potential hazard of this treatment. PMID- 8235436 TI - Structural and morphometric analysis of murine small intestine after indomethacin administration. AB - Indomethacin, a nonsteroidal anti-inflammatory drug, induces the formation of gastrointestinal ulceration both in experimental animals and in humans. A study of indomethacin-induced ulcers in the mouse showed that two doses of indomethacin, each administered subcutaneously at 85 mg/kg body weight, induced well-defined gastrointestinal ulcers in C57 mice, accompanied by inflammatory and vascular changes in the stomach and small intestine. Maximal damage was observed 20 h after the second dose of indomethacin. Morphometric analysis identified changes in all compartments of the small intestine. There was a marked reduction in the length of the small intestine, intestinal dilatation, a significant decrease in villous height, with the formation of subepithelial blisters or blebs within villi, and submucosal vascular dilatation. There was no change in the number of villi or of submucosal arterioles or in the total amount of muscle present in the wall of the intestine. The tissue changes identified in this study may have implications for gut function at specific periods during indomethacin treatment. PMID- 8235437 TI - Cholesterol nucleation time measurement in nasobiliary or nasoduodenal bile. Comparison with surgical bile. AB - The usual technique of collecting gallbladder bile at laparotomy is not suitable for sequential studies of cholesterol nucleation time (NT) in patients receiving therapy to prevent or dissolve cholesterol gallstones. Our aim was to study the feasibility of measuring NT in bile obtained by nasobiliary or nasoduodenal intubation. We studied a total of 10 cholesterol gallstone patients; in 8 bile was collected by nasobiliary drainage, in 7 it was collected by nasoduodenal intubation, and in 3 it was collected at laparotomy the next day. Three patients developed abdominal pain and increased serum amylase after endoscopic retrograde cannulation. All three biles obtained at operation nucleated quickly (NT, 1-4 days), whereas duodenal biles were all beyond the expected range (NT, > 21 days). Chymotrypsin activity, as a marker of pancreatic juice contamination, was detected in five of eight nasobiliary biles and in all seven duodenal biles but in none of the surgical biles. Free fatty acids (reflecting lipolysis) were significantly higher in duodenal than in surgical biles, with nasobiliary bile showing intermediate values. Nasobiliary bile showed either a rapid (median NT, 3 days) or a slow (median NT, 22 days) NT, depending on whether chymotrypsin activity was absent or present (p < 0.05). We conclude that duodenal bile is never suitable for NT determination because of contamination by pancreatic enzymes, and that nasobiliary bile, if not contaminated by pancreatic enzymes, may be suitable for NT determination but that its collection via a nasobiliary tube after cholecystokinin injection carries a risk of pancreatitis. PMID- 8235438 TI - Diurnal profile of oxyntomodulin-like immunoreactivity in duodenal ulcer patients. AB - Plasma concentrations of oxyntomodulin-like immunoreactivity, a group of intestinal peptides capable of mediating an enterogastrone signal, were measured during a 24-h period in 6 duodenal ulcer patients and compared with those of 16 age-matched controls. Each subject was submitted to 18 oxyntomodulin-like immunoreactivity determinations. Four standardized meals were given during the test. Furthermore, each patient was evaluated for peak acid output after pentagastrin stimulation. The values of the duodenal ulcer subjects were predominantly within normal acid secretion limits. Fasting levels, meal-induced variations, and nocturnal production of oxyntomodulin-like immunoreactivity were similar in the two groups. A negative correlation was observed between peak acid output and oxyntomodulin-like immunoreactivity evaluated either as nocturnal production or as maximum nyctohemeral concentration. We conclude that, taken as a whole, duodenal ulcer disease is not caused by a defect in oxyntomodulin-like immunoreactivity secretion. However, this study does not rule out the possibility of a selective deficiency of these peptides in some duodenal ulcer subgroups such as hypersecretory patients. PMID- 8235439 TI - Treatment of ulcerative colitis with high doses of oral prednisolone. The rate of remission, the need for surgery, and the effect of prolonging the treatment. AB - Treatment of acute attacks of ulcerative colitis in 89 patients with doses of prednisolone above or equal to 40 mg resulted in an overall remission in 67%. Remission rate and colectomy rate were 47% and 42%, respectively, when the disease was severe, 80% and 13% when moderate, and 84% and 3% when mild. The need for surgery was 28% in pancolitis, 11% in left-sided colitis, and 5% in proctitis. After subsequent treatment episodes colectomy was performed in 35% of patients with pancolitis, in 37% with left-sided colitis, and in 5% with proctitis. The median total duration of therapy in patients who went into clinical remission was 4 months, and the median dose just above 3 g prednisolone. Patients who stayed in remission during the follow-up received a significantly higher start dose and total dose of prednisolone in the treatment episode than patients who had a relapse. In 25 patients treatment with doses equal to or above 75 mg of prednisolone was continued beyond 10 days, and 11 patients experienced remission whereas 14 patients had surgery performed. Orally administered corticosteroids produce results comparable to those obtained after the previously suggested intravenous regimen. PMID- 8235440 TI - Changes in oesophageal pH associated with gastro-oesophageal reflux. Are traditional criteria sensitive for detection of reflux? AB - Traditionally, gastro-oesophageal reflux is deemed to have occurred when oesophageal pH falls below 4. Other 'non-traditional' pH changes that do not fall below pH 4, that fall below 4 for only brief intervals, or that occur when basal pH is less than 4 are usually disregarded. The aim of this study was to determine whether these non-traditional pH changes represent gastro-oesophageal reflux or are artefactual. The 3-h postprandial combined oesophageal pH and manometric records of 22 patients referred for investigation of suspected gastro-oesophageal reflux were reviewed. All pH falls of > or = 0.5 pH units were analysed for manometric evidence of reflux that was classified as definite, probable, or possible. In total, 196 traditional and 223 non-traditional pH events were scored and analysed. The majority of traditional (80%) and non-traditional (60%) events were associated with definite manometric evidence of reflux, although a greater proportion of non-traditional events were associated with only probable evidence of reflux (33%) compared with traditional events (18%). The proportions of possible reflux were similar in the two groups. Limiting pH events to only those satisfying traditional criteria excluded an additional 32% with definite manometric evidence of reflux and 49% with definite or probable evidence of reflux. Most pH falls that remained above 4 or fell across 4 for < 15 sec occurred in the 1st h postprandially, compared with traditional pH events, which occurred equally throughout the 3-h period. We conclude that traditional criteria for scoring pH episodes substantially underestimate the number of reflux episodes. PMID- 8235441 TI - Effects of dibutyryl cyclic AMP and papaverine on intrahepatocytic bile acid transport. Role of vesicle transport. AB - The secondary messenger cyclic AMP plays an important role in regulating biliary excretory function by stimulating the transcytotic vesicle transport system, whereas papaverine exerts an inhibitory effect on this system. We therefore investigated their effects on bile acid-induced cytotoxicity and intrahepatocytic content of bile acid in primary cultured rat hepatocytes. Simultaneous addition of 1 mM dibutyryl cyclic AMP (DBcAMP), an analogue of cAMP, with 1 mM taurochenodeoxycholic acid (TCDCA) significantly decreased the release of lactate dehydrogenase (LDH) as compared with the case with 1 mM TCDCA alone (7.1 +/- 0.13% of total versus 10.7 +/- 0.3%). In contrast, 0.1 mM papaverine approximately doubled the amount of LDH (22.0 +/- 0.6% of total versus 10.7 +/- 0.3%; P < 0.01). The intracellular content of TCDCA 180 min after the administration of 1 mM TCDCA alone was 20.8 +/- 0.7 nmol/mg protein, that after simultaneous administration of 1 mM DBcAMP, 16.2 +/- 1.0 nmol/mg protein, and that after the simultaneous administration of 0.1 mM papaverine, 38.5 +/- 1.9 nmol/mg protein. A clear correlation between the release of LDH from hepatocytes and the intracellular content of TCDCA was thus observed. When given together with 1 mM taurocholic acid (TCA) or 1 mM tauroursodeoxycholic acid (TUDCA), papaverine exerted little effect on cytotoxicity or intrahepatocytic bile acid content. When cells were bathed in a medium free of bile acid after pretreatment with 1 mM TCDCA and 1 mM DBcAMP, additional exposure to DBcAMP for 30 min significantly stimulated reduction of intracellular TCDCA content (30.2 +/- 0.4% of total versus 44.0 +/- 1.4%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235442 TI - Macrophage-mediated enterocyte damage in BALB/c mice infected with different strains of Giardia lamblia. AB - The mechanism of mucosal injury in Giardia lamblia-infected animals and humans is not well understood, although the role of gut macrophages in killing the trophozoites is well known. It is speculated, however, that macrophage products have a role in tissue injury and inflammatory response during infection, as in other inflammatory diseases. Therefore, in the present study an attempt was made to examine the mechanism involved in enterocyte damage during giardiasis. This was achieved using co-culture of enterocytes and gut macrophages obtained from infected BALB/c mice. The extent of tissue damage was assessed by measuring the marker enzyme of enterocyte damage, lactate dehydrogenase. To investigate the role of the various proteases and free oxygen radicals released by activated macrophages on enterocyte damage, inhibitors of various proteases and free oxygen radicals were used. Superoxide radical and certain proteases were found to have important roles in bringing about enterocyte damage during this infection in mice. Parasite load, lactate dehydrogenase release, and extent of lipid peroxidation were more pronounced in mice infected with symptomatic strains than in asymptomatic ones. The theory of inflammatory cell-mediated enterocyte damage in Giardia lamblia infection is proposed. PMID- 8235443 TI - Characterization of the joining chain (J-chain) promoter. AB - A 300 b.p. promoter from the mouse joining chain (J-chain) gene was studied with regard to functional activity and protein/DNA interactions. The promoter only stimulated expression of a chloramphenicol-acetyl-transferase (CAT) reporter gene when an enhancer was present in the construct, regardless of whether the construct was transfected into cell lines that did or did not express an endogenous J-chain. Furthermore, deletion mutants lacking the 5' portion of the promoter were transcribed at a higher rate than the intact promoter in both J chain positive and J-chain negative B-cell lines but not in untransformed B lymphocytes stimulated by lipopolysaccharide, indicating the presence of a negative control element in the 5' portion of the J-chain promoter active in tumour cells only. The octamer element in the J-chain promoter was found to bind Oct proteins, albeit with a low affinity. The penta-deca (p.d.) element in the J chain promoter bound proteins in extracts from untransformed B cells but not in the tested cell lines. The protein binding to the J-chain p.d. element did not compete efficiently with a p.d. element from the SP6 kappa promoter. A protein binding to the 5' portion of the J-chain was expressed in some cell lines but not in others; neither a negative nor a positive correlation to J-chain expression could be seen. It was concluded that the J-chain promoter is equivalent to a kappa promoter and that differentiation-specific J-chain expression is governed by distal, positive control elements located outside the analysed region. PMID- 8235444 TI - Differential expression of rabbit serum amyloid A genes in response to various inflammatory agents. AB - Serum amyloid A (SAA) is an acute-phase plasma protein which increases up to 1000 fold after an acute-phase stimulus. Several SAA genes and corresponding protein isotypes exist in individual species. Liver is the main source of production, but extra-hepatic SAA expression has been described. In this study inflammation was induced in rabbits with lipopolysaccharide, turpentine, or casein. Transcription of SAA mRNA was studied using Northern blot analysis with probes specific for three different rabbit SAA isotypes and analysed by scanning densitometry. In the stimulated liver slight variation in SAA mRNA transcription level was seen after stimulation with different inflammatory agents. After lipopolysaccharide stimulation SAA gene expression was also seen in most of the extra-hepatic organs. After turpentine stimulation SAA mRNA was seen in the liver, the ovary, and the small intestines, and after casein stimulation only in the liver and the ovary. SAA1 and SAA2 were induced exclusively in the liver, while SAA3 was induced mainly in the extra-hepatic organs. This indicates that the SAA genes probably are independently regulated both in relation to stimulus, gene- and tissue-specificity. PMID- 8235445 TI - Impaired phagolysosomal fusion of peripheral blood monocytes from HIV-infected subjects. AB - We evaluated phagolysosomal fusion in peripheral blood monocytes from 20 HIV infected individuals and 40 normal controls, using a fluorescence assay with acridine orange as marker. The percentages of phagolysosomal fusion of monocytes from HIV-infected subjects, after 30 and 60 min of yeast ingestion, (mean +/- standard deviation) 57.2 +/- 17 and 63.2 +/- 18.6, respectively, when compared to normal controls (72.4 +/- 7.8 and 77 +/- 8.1), did not differ significantly. However, there was a direct linear association between the percentages of phagolysosomal fusion and CD4+ lymphocytes (P < 0.001) or CD4/CD8 T-cell ratio (P < 0.01). These results suggest that phagolysosomal dysfunction becomes evident at late stages of HIV infection and progresses as CD4+.T-lymphocyte count and CD4/CD8 T-cell ratio decrease. On the other hand, recombinant gp120 inhibited significantly normal phagolysosomal fusion at concentrations ranging between 1 and 1000 ng/ml. Taking together the results obtained, we can conclude that gp120 could be responsible for monocyte phagolysosomal dysfunction observed in HIV infected patients. PMID- 8235446 TI - Characteristics of IgA antibodies against HIV-1 in sera and saliva from HIV seropositive individuals in different clinical stages. AB - IgA antibodies were analysed in sera and saliva from 40 HIV-1 seropositive individuals. The level of total IgA in serum was elevated according to the progress of the disease. IgA antibodies against p24 and gp160 were detected in the asymptomatic phase of infection. However, they declined in the symptomatic phases in contrast with IgG antibodies. Interestingly, three patients in the symptomatic phase who showed high levels of IgA antibodies were all in relatively good clinical condition. The IgG and IgA antibodies in saliva declined in the symptomatic phase. The level of IgG anti-p24 antibodies in saliva correlated with that in serum, suggesting that IgG anti-p24 antibodies in saliva originated from those in the serum. These results indicate that IgA antibodies are regulated independently from IgG antibodies and that the mucosal immune system is impaired early in the symptomatic phase of HIV infection, which starts with mucosal impairment. Detection of IgA antibodies may be useful for prognosis of the disease in HIV-infected individuals. The results indicate also that treatment for the impaired IgA mucosal immune system should be taken into consideration. PMID- 8235447 TI - Serum amyloid-P component of the Armenian hamster: gene structure and comparison with structure and expression of the SAP gene from Syrian hamster. AB - Serum amyloid P (SAP), a phylogenetically conserved pentraxin, is an integral component of all amyloid deposits. Regulation of expression of SAP gene expression is quite different in two related hamster species. In Syrian hamsters, the resting serum levels of SAP are determined by gender, and the direction of alteration following inflammation is divergent. In Armenian hamsters, SAP is not a prominent acute-phase reactant and there is no gender dimorphism of expression. The structure and expression of the SAP gene of the Armenian hamsters was investigated by isolation of genomic clones, nucleotide sequence analysis, and RNA studies. The gene structure of Armenian hamster SAP is similar to the genes of all other pentraxins studied. While the upstream regions of the SAP genes of Syrian and Armenian hamsters are quite similar, important differences in potential enhancer sites have been recognized by comparing the corresponding sequences of SAP genes from both species. Little alteration in hepatic levels of transcripts encoding SAP or CRP, the other pentraxin, were noted following administration of lipopolysaccharide to Armenian hamsters. This relative lack of response occurred despite a marked acute phase reaction documented for serum amyloid A mRNA levels. PMID- 8235448 TI - Do human Peyer's patches contribute to the intestinal intraepithelial gamma/delta T-cell population? AB - T-cell receptor gamma/delta+) (TcR gamma/delta+) lymphocytes in human Peyer's patches (PP) adjoining ileal mucosa were studied by monoclonal antibodies with paired immunofluorescence staining in situ and by flow-cytometric phenotyping of isolated cells. The proportion of gamma/delta+ T cells in the follicle-associated epithelium outside the M-cell areas (median 4.1%, range 2.2-30.1%) was similar to that in mucosal villous epithelium (median 4.4%, range 0.5-30.5%). Most intraepithelial gamma/delta cells (approximately 90%) expressed CD45R0 but only a few expressed CD8 (< 10%) and none L-selectin; a dominating subset (median 46%) employed the V delta 1/J delta 1 gene product (range 22-100%). The M-cell areas lacked gamma/delta cells but contained clustered CD20+ and CD3+ lymphocytes. The subepithelial PP dome area and interfollicular (T-cell) zones, as well as the mucosal lamina propria, contained very few gamma/delta cells (median 1.7%, range 0.4-8.9%) which were dominated (88-100%) by the V delta 2-encoded subset. Those in the dome area and lamina propria were often (approximately 75%) CD45R0+ (range 44-90%) while very few (approximately 2%) expressed L-selectin (range 0-15%). By contrast, CD45R0 expression on gamma/delta cells in the PP T-cell zones was relatively low (approximately 46%) and that of L-selectin relatively high (approximately 43%). In conclusion, TcR gamma/delta + cells are quite rare in human PP and belong mainly to the V delta 2-encoded subset, thus being different from most intraepithelial gamma/delta cells that probably have another origin. The L-selectin+ fraction of PP gamma/delta cells presumably represent newly recruited 'naive' T lymphocytes while CD45R0+ gamma/delta cells both in PP and lamina propria are probably antigen-primed. PMID- 8235449 TI - Polyclonal activation of B-lymphocytes and induction of autoimmunity in retrovirus infected NMRI mice. AB - Polyclonal activation of B-lymphocytes accompanies many retroviral infections. Friend derived murine immunosuppressive virus (Fd-MIV) is a non-defective murine retrovirus which was isolated from T-helper cells from mice infected with the acute transforming retrovirus Friend leukaemia complex (FLC). In contrast to FLC, Fd-MIV does not induce acute transformation of lymphoid and erythroid cells but causes immunosuppression and lymphadenopathy in adult NMRI mice. The effect of Fd MIV infection on B-lymphocytes was studied. Fd-MIV infection led to a persistent hypergammaglobulinemy with a significant increase in the level of circulating IgG, IgM and immune complexes. In the spleen and lymph nodes, B-lymphocyte proliferation was found. Parallel to the development of hypergammaglobulinemy, autoantibodies to a variety of nuclear and other autoantigens was detected. In conclusion, the Fd-MIV infection leads to a B-lymphocyte dysfunction that has many parallels with AIDS. Furthermore, the Fd-MIV infection seems to represent an example of an autoimmune condition caused by an exogenous infectious agent. PMID- 8235450 TI - The detection and biological activity of human antibodies to IL-2 in normal donors. AB - Antibodies to interleukin-2 (IL-2) have been reported in the sera of patients under therapies involving this cytokine and in the sera of patients infected with human immunodeficiency virus (HIV). Our study proves that auto-antibodies to IL-2 are present also, at a lower titre, in healthy individuals. These antibodies were affinity purified and studied for their capability to interfere with the in vitro biological activity of IL-2. Data obtained show that human anti-IL-2 antibodies can interfere with lymphocyte proliferation both in the lymphokine activated killer (LAK) cell assay and in the mixed lymphocyte reaction (MLR). However, the kinetics of inhibition by anti-IL-2 antibodies differs from LAK cell assay to MLR as the former are always inhibited in a time-independent manner, and the latter only by adding antibodies at the outset of culture. The neutralizing activity observed in vitro suggests that such antibodies play a part in the elaborate cytokine network by which the immune system regulates the amplitude and duration of its response. PMID- 8235451 TI - Immune modulatory effects of immunoglobulins on cell-mediated immune responses in vitro. AB - Intravenous Immunoglobulin (IVIG) at a concentration of 5 mg/ml, significantly inhibited mitogenic responses to phytohaemagglutinin (PHA), concanavalin A (conA) and pokeweed mitogen (PWM) by peripheral blood cells from healthy donors. No difference in inhibition by IVIG was seen when stimulating different T-lymphocyte cell subsets. Inhibition by IVIG was dose-dependent. An increased response was observed when IVIG was added more than 12 h after PHA compared to adding 1 h before [P = 0.05]. Intravenous immunoglobulin added to mixed lymphocyte cultures (MLC), reduced the median response by more than 60% (range 14-89%; P = 0.03) and almost completely abrogated the lymphocyte response to Staphylococcus aureus protein A (SPA), whose median inhibition was 94% (range 90-99%; P = 0.02). When comparing 12 different commercial IVIG preparations at a concentration of 2.5 mg/ml, the median inhibition of the PHA stimulation ranged from 4% to 35% and the MLC response from 0% to 66%. In the presence of IVIG the lymphocyte response to different herpes virus antigens was reduced by > 50%. No difference in inhibitory effect was seen when comparing IVIG and cytomegalovirus (CMV) hyper Ig, but CMV negative Ig resulted in lower inhibition [P = 0.05]. Three out of five IgG preparations (2.5 mg/ml) made from single donors inhibited PHA stimulation significantly more than commercial IVIG [P < 0.05]. Mean inhibition was 61% compared to 35%. Inhibition by pooled IgG from five donors was 56%. F(ab')2 fragments of IVIG inhibited the MLC response by more than 50% (range 34-75%), SPA stimulation by 97% (83-104%) and PHA stimulation by more than 30% (26-37%). One of two Fc preparations tested had an inhibitory effect, but the inhibition was less than that obtained with the F(ab')2 fragments [P = 0.04]. These results further strengthen the notion that IVIG exerts its immune modulatory effect by binding to leukocyte surface receptors. A clear inhibition was obtained with concentrations corresponding to the serum levels obtained when IVIG is given 250 500 mg/kg bodyweight. F(ab')2 fragments have the same inhibitory effect as intact IgG molecules but the role of Fc fragments still remains unclear. Differences in the immunosuppressive effect of various IVIG preparations may be associated with the method of preparation. PMID- 8235452 TI - Bacterial lysates and ribosomes as inducers of specific immune responses: a comparative study. AB - A bacterial lysate (OM-85 BV), a preparation of purified bacterial ribosomes (D53) and a placebo were tested for ability to induce the local appearance of specific antibody-containing cells. The three compounds were given orally to 90 children who required tonsillectomy. Surgery was carried out after 1 month of therapy. Frozen-cut sections of each tonsil were tested in indirect immunofluorescence. Cells containing antibodies directed to Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae or Klebsiella pneumoniae were enumerated. Lowest values were noted in the placebo group. Slightly higher numbers were observed after treatment with OM-85 BV, but significant increases were noted only for the elevated numbers of specific antibody-containing cells observed after D53 therapy. Bacterial ribosomal preparations thus contribute efficient induction of specific local immune responses in man. PMID- 8235453 TI - Herpes simplex encephalitis. Early diagnosis and immune activation in the acute stage and during long-term follow-up. AB - From a series of in all 93 patients with herpes simplex encephalitis (HSE), verified by biopsy and/or the demonstration of intrathecal synthesis of antibodies to the virus, cerebrospinal fluid (CSF) and serum samples were analysed and compared with samples from 80 patients with non-HSE, i.e. acute encephalitis of non-HSV origin (approximately 50% with other known aetiology, 50% of unknown origin) treated on the suspicion of HSE but in whom no signs of intrathecal HSV antibody synthesis were found, and samples from an additional 42 patients with other verified or suspected diseases of the CNS. To improve the early non-invasive diagnosis of HSE, a HSV IgG capture enzyme linked immunosorbent assay (ELISA) was developed to demonstrate intrathecal synthesis of antibodies to the virus and the results were compared to those of the indirect ELISA. The capture ELISA was found to be advantageous in detecting the early antibody response and yielded more clear-cut results. No correction for damage to the blood-CSF barrier was needed and the method was therefore less labour intensive than the indirect ELISA. Furthermore, a polymerase chain reaction (PCR) assay, with two "nested" primers pairs selected in the glycoprotein D gene of HSV 1, was developed for the amplification of HSV DNA in CSF. The method was found to be a rapid and non-invasive means of diagnosing HSE in a very early stage of the disease; it was highly sensitive and specific. With a combination of nested PCR assays for HSV-1 and HSV-2 (primers in the glycoprotein G gene) in 10 microliters of CSF, HSV DNA was detected in CSF from 88 out of 93 patients (95%) with HSE. Evidence of HSV-2 aetiology was found in 6 of 93 consecutive cases of HSE in immunocompetent patients by type-specific assays for the demonstration of HSV-2 DNA (primers in the gG gene) and HSV-2 antibodies (to gG2 antigen) in the CSF. Five of the 6 patients with HSV-2 encephalitis exhibited a clinical picture of severe HSE indistinguishable from that of "classical" HSV-1 encephalitis. The combined use of PCR for the detection of HSV DNA in the CSF and the demonstration of intrathecal synthesis of antibodies to the virus will yield a reliable diagnosis and is now the method of choice for the diagnosis of HSE.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8235454 TI - Contagion in schizophrenia: a critique of Crow and Done (1986). AB - This article discusses the literature on the possible role of viruses in the development of schizophrenia and outlines the evidence that compelled Crow and Done (1986) to reject a horizontal contagion hypothesis (e.g., sibling to sibling transmission). We posit a genetically determined age after which one becomes vulnerable to the illness, rather than a strict age of onset. We also propose an environmentally determined range of resistance to this susceptibility as an alternative conceptualization of the role of inheritance and of viral exposure in the etiology of the disease. The predictions derived from our new model fit the findings of Crow and Done and are consistent with the older literature of possible viral factors, thus reaffirming the horizontal contagion hypothesis. PMID- 8235455 TI - Eye tracking dysfunction and schizophrenia: a critical perspective. AB - Eye tracking dysfunction (ETD) has been found in large numbers of schizophrenia patients and their first-degree relatives. Because of the many replications of the central findings, ETD has been proposed as a useful way of expanding the schizophrenia phenotype in genetic studies. We critically review the literature on ETD with respect to issues of measurement and the search for quantitative indices of ETD; syndrome and familial specificity of ETD for schizophrenia; statistical, interpretive, and methodological considerations in the use of mixture analysis; the association of ETD with clinically and psychometrically defined schizotypy; and the questions of trait stability and medication effects. PMID- 8235456 TI - Neurobiological investigations in cingulate cortex of schizophrenic brain. AB - Several studies have indicated that the pathophysiology of schizophrenia may involve discrete alterations of intrinsic circuits within the anterior cingulate cortex, particularly in layer II. These findings have suggested the possibility that a disturbance in neuronal migration or differentiation during the perinatal period could be related to these differences in schizophrenia subjects. To explore the idea that schizophrenia may be a neurodevelopmental disorder in which precise changes in synaptic connectivity occur, model circuits can be used as empirical tools for identifying the nature and origin of layer II abnormalities in the cortex of schizophrenic brain. Eventually, a strategy of this type may point the way to novel treatment approaches that will limit or perhaps even prevent the deterioration that occurs in patients with schizophrenia. PMID- 8235457 TI - Minor physical anomalies in schizophrenia and mood disorders. AB - The Waldrop Physical Anomaly Scale was used to assess the prevalence of minor physical anomalies in three groups: schizophrenia patients (n = 118), patients with mood disorders (n = 33), and normal controls (n = 31). Patients with schizophrenia had significantly more anomalies than controls. Patients with mood disorders did not have significantly different anomaly scores than schizophrenia patients or controls. Patients with tardive dyskinesia (TD) had significantly more anomalies than those without TD. Physical anomalies in the schizophrenia group were not found to be related to severity of psychopathology, age of onset, positive or negative schizophrenic symptoms, or socioeconomic status. PMID- 8235458 TI - Seasonality of schizophrenia and stillbirths. AB - A study of the birth pattern of 30,467 patients with schizophrenia and 428,406 stillbirths in New York State showed a significant winter-month excess in both schizophrenia births (p = 0.0000) and stillbirths (p = 0.0000). Excess births of individuals with schizophrenia in November and December provided additional evidence to refute the age-incidence explanation for the observed seasonality. Time series spectral analysis showed coherence in the pattern of schizophrenia births and stillbirths of 0.898 (p < 0.003) with schizophrenia births preceding stillbirths by 13 days. These results suggest that a common etiological seasonal factor affects a subgroup of births of individuals who later develop schizophrenia and a subgroup of stillbirths. Nutritional factors, temperature, and infectious agents known to cause both central nervous system disease and stillbirths are possible candidates. PMID- 8235459 TI - Childhood precursors of affective vs. social deficits in adolescents at risk for schizophrenia. AB - Childhood attentional and neuromotor precursors of social competence and affective deficits in adolescents at risk for schizophrenia, adolescents at risk for affective disorder, and matched comparison adolescents were examined. The subjects were offspring of parents with schizophrenia or affective disorder and of normal parents matched on age, sex, and socioeconomic status from the New York High-Risk Project (Sample B). On the basis of interviews conducted when the subjects were children and adolescents, social competence was rated from child reports and parent reports, affective deficits were assessed by affective flattening ratings, and smiling was assessed by counting broad smiles. Adolescents at risk for schizophrenia had significantly greater social and affective deficits than adolescents at risk for affective disorder and comparison adolescents. In subjects at risk for schizophrenia, childhood neuromotor dysfunction predicted adolescent affective flattening, and childhood attentional dysfunction predicted adolescent social deficits. The results suggest that affective and social deficits in schizophrenia have different childhood precursors. PMID- 8235460 TI - Schizophrenic delusions: a phenomenological approach. AB - The issue of specificity of delusions in schizophrenia is still a matter of debate. The authors analyze the delusion formation in schizophrenia from a prototypical, phenomenological point of view, focusing on the subject's experience. This perspective links delusion formation to the autistic predisposition, which is considered here as the elementary phenotypic expression of the vulnerability to schizophrenia. Autism is viewed as a defective preconceptual (i.e., before language) attunement to the world. It impedes the individual's sharing of "common sense" with others and impairs the ability to project into the future. The development of delusions is illustrated, in part, by Klaus Conrad's work on the onset of paranoid schizophrenia. Delusions are viewed as transformations of the structure of experiencing. When threatened in future ability to be, the autistic, vulnerable person looks for the clues to becoming by attributing significance to disparate elements of the environment, which become self-referential. The link established between these disparate elements is based on universal characteristics that give the schizophrenic delusion a metaphysical quality. The transitivistic experience in delusions of control and omnipotence points to a specific way of crossing the border between "mine" and "yours" (disturbances of the experiencing "I"). What strikes a clinician in these delusions is that the normally tacit link between the sense of being and the sense of acting becomes quite apparent. The authors also propose a specificity in the themes of schizophrenic delusions. Delusions acquire a schizophrenic quality when ontological (i.e., universal) elements of the discourse between the locutor and the Other dominate at the expense of the worldly elements. It is emphasized that delusional content and form are dialectically related and hardly distinguishable. The authors consider the delusion formation as a phenomenon of emergence, a situation in which a new qualitative order arises from the reorganization of essentially unchanged elements. To consider schizophrenia as an emergent, particular way of experiencing, related to the autistic defect, has important consequences for research and for treatment. A dialectic exchange is needed between prototypical models generated by phenomenological inquiry and empirical, operational validation of testable aspects of such models. PMID- 8235461 TI - Psychosocial functioning and subjective experience in schizophrenia. AB - This study examines the relationship between psychosocial functioning and subjective experience in 193 individuals diagnosed with schizophrenia or schizoaffective disorder according to Research Diagnostic Criteria. Psychosocial functioning was measured as work functioning, social functioning, living situation, symptomatology, and intrapsychic aspects of the deficit syndrome. Subjective experience consisted of measures of self-esteem, satisfaction with life, and subjective distress. Multivariate analyses resulted in two major findings. The first finding is a model of psychosocial functioning that consists of two factors: disorder-related variables (symptomatology and intrapsychic deficits) and functional status variables (work, social, and living situation). The second major finding is a two-tiered model of the relationship between psychosocial functioning and subjective experience. The model suggests a primary and pervasive relationship between the disorder-related variables and subjective experience and a secondary and less pronounced relationship between functional status variables and subjective experience. The implications of these findings for treatment and rehabilitation and for the study of subjective experience in schizophrenia are discussed. PMID- 8235462 TI - Determinants of psychological distress in relatives of people with chronic schizophrenia. AB - Research shows that the relatives of people with chronic schizophrenia suffer psychological distress and interference in daily life from the burdens of caregiving. In this study, members of a self-help group for the relatives of schizophrenia sufferers provided information through a mail survey, using standardized measures of psychological distress and burden, and severity of the sufferer's illness. Caregiver psychological distress was high compared with test norms, and the level of behavioral disturbance in the sufferer was found to contribute to caregiver distress after controlling for the caregiver's age, sex, and social supports. An unexpected finding was that those caring for female sufferers reported greater distress than those caring for males. PMID- 8235463 TI - Negative symptom assessment of chronic schizophrenia patients. AB - A new scale for assessing negative symptoms in schizophrenia, the Negative Symptom Assessment (NSA), was administered to 101 male chronic, inpatient schizophrenia patients. Factor analysis of the NSA yielded seven factors, but most of the explained variance resided in Factor 1, Restricted Affect/Emotion. The factors that emerged from this study closely resembled NSA factors derived from an earlier study of outpatient schizophrenia patients, which indicates the factor structure of the NSA is robust. A constellation of variables reflecting long-term or chronic illness were significantly related to six of the seven factors. These results suggest that "institutionalism" may play a role in the evolution of some negative symptoms. PMID- 8235464 TI - Sibling perspectives on schizophrenia and the family. AB - Intensive audiotaped interviews with 14 adult siblings of schizophrenia patients were used to explore their experiences. Interviews focused on the ways adult siblings define and experience their sibling's mental illness and their participation in illness management. Cross-case analysis revealed that recognition of schizophrenia in a brother or sister was characterized by a pivotal, reorganizing event that redefined behavior previously viewed as difficult or different. Subjects' approach to the sibling's illness reflected three distinct patterns: collaborative, crisis-oriented, and detached. The impact of a mentally ill brother or sister on subjects' lives ranged from pervasive to discrete and was seen as emanating from the family and the sibling, but in different ways. These findings can help sensitize clinicians to the complex nature of sibling experiences as well as to the needs of siblings and their families. PMID- 8235466 TI - [Legionella II. New contributions to the evaluation of hygienic problems]. PMID- 8235465 TI - First person account: the children of madness. PMID- 8235467 TI - [Incidence and evaluation of Legionella in hospitals and other large buildings]. PMID- 8235468 TI - [Clinical aspects and therapy of legionella infection]. PMID- 8235469 TI - [Occurrence of Legionella in drinking water supply facilities in single and two family houses]. PMID- 8235470 TI - [The DVGW professional regulations W 551]. PMID- 8235471 TI - [Further technical regulations and the Legionella problem]. PMID- 8235472 TI - [Reduction of Legionella in shower water of indoor pools. UV disinfection as an alternative to temperature increase?]. PMID- 8235473 TI - [Comments on the multiplication and eradication temperatures of Legionella]. PMID- 8235474 TI - [Prevention of Legionella multiplication in swimming and whirlpools]. PMID- 8235475 TI - [Legionella in dentistry equipment]. PMID- 8235476 TI - [Results of chemical and microbiological studies of indoor climate control equipment in Berlin]. PMID- 8235477 TI - [Indoor climate control equipment. Requirements for microbiocide additives to humidifier water]. PMID- 8235478 TI - [Diagnosis of legionellosis--current status]. PMID- 8235479 TI - [Incidence and significance of Legionella in industrial air conditioning systems]. PMID- 8235480 TI - [Practical experience with control of Legionella in cold water circulation systems by water conditioning measures]. PMID- 8235481 TI - [Environmental health aspects for control of legionellosis. Recommendations of the World Health Organization (WHO) Regional Office for Europe, Copenhagen 1986]. PMID- 8235482 TI - [Comments on the problem of Legionella in Germany]. PMID- 8235483 TI - [Epidemiology of legionellosis]. PMID- 8235484 TI - [Netherlands' experience with legionellosis outbreaks]. PMID- 8235485 TI - [Surveillance and control of Legionnaires' disease in England and Wales]. PMID- 8235486 TI - [Temperature layers and their effects on serial water heaters]. PMID- 8235487 TI - [Implementing the federal communicable disease regulation]. PMID- 8235488 TI - [Technical prerequisites of equipment for hygienically warmed water]. PMID- 8235489 TI - IgE antibodies specific for cartilage collagens type II, IX and XI in rheumatic diseases. AB - Serum samples from 149 patients with RA and other rheumatological diseases, and 57 non-arthritic controls have been assayed for IgE antibodies to the cartilage collagen types II, IX and XI in their native and denatured state. Using an improved ELISA technique together with antigen-binding inhibition studies to confirm specificity, 10 of the 149 (7%) patients showed IgE antibodies to human collagen type II and bovine collagen types II, IX or XI. Some patients responded to only one collagen type whereas others had IgE positive responses to two or all three collagen species. Most of the IgE responses detected were directed towards the denatured collagens. Those sera showing an IgE response to bovine type II collagen produced a similar response to the human equivalent, including two patients with SLE. None of 57 control subjects demonstrated IgE specificity for any of the cartilage collagens. Patients with IgE specificity for the cartilage collagens did not demonstrate IgM or IgA specificity for these antigens, but two of these patients showed IgG responses to type II and XI collagens. Whereas eight patients were exclusively IgE-positive for the cartilage collagens, others expressed specificities for IgG, IgM or IgA. It therefore appears that the specific autoimmune profile for each patient is often different from others, both in terms of the class of immunoglobulin expressed and the collagen antigens recognised. At present no correlations were observed between the IgE-positive patients and their clinical assessment and/or prognosis. PMID- 8235490 TI - Serum levels of soluble interleukin-2 receptor in patients with systemic lupus erythematosus and systemic idiopathic vasculitis. AB - Soluble interleukin 2 receptor (sIL2R) may be used as an index of immune perturbation. We report on the correlation between the serum levels of sIL2R, as assessed with a sandwich-ELISA, and disease activity in 61 patients with Systemic Lupus Erythematosus (SLE) and in 15 with systemic idiopathic vasculitis (SIV). The mean levels of sIL2R in SLE and SIV patients were significantly higher than in healthy controls and higher values were detected in patients with active disease or severe organ involvement or infection. We also studied patients with clinically silent SLE, characterized by the presence of several immunologic abnormalities. The sIL2R mean level in this group did not differ from that of quiescent SLE patients, suggesting that immunologic alterations are present even in inactive SLE. Finally, the sIL2R assay showed higher sensitivity and specificity than most of the common immunologic parameters. PMID- 8235491 TI - The prognostic value of HLA DR4 and B27 antigens in early rheumatoid arthritis. AB - The prognostic significance of HLA DR4 and B27 antigens was investigated in a 3 year follow-up of 87 patients with early rheumatoid arthritis (RA). The frequencies of DR1, DR4 and also of B27 were increased and the frequencies of DR2, DR3 and DR7 decreased compared with the normal Finnish population. During the follow-up with antirheumatic treatment, a similar improvement in clinical variables and laboratory measure assessing disease activity was found in both DR4 positive and DR4-negative RA patients. Despite clinical improvement a fast radiological progression in peripheral joints was observed but the presence of DR4 or B27 had no impact on the progression of joint damage. In some patients cervical changes developed early in the course of RA but were not related to DR4 or B27 positivity. The earlier observation of increased prevalence of HLA B27 in the Finnish RA patients was confirmed but the presence of B27 did not modify the clinical picture of RA. PMID- 8235492 TI - Lung involvement in rheumatoid factor-negative arthritis. AB - Bronchoalveolar lavage (BAL) was performed on 13 asymptomatic patients with sero (rheumatoid factor)-negative arthritis (SNA); (6 with peripheral psoriatic arthritis, 2 with axial psoriatic arthritis, 3 with ankylosing spondylitis, 2 with sacroiliitis). BAL revealed a significant decrease of neutrophil granulocytes and an increase of B-lymphocytes in patients with SNA in comparison with 64 patients with rheumatoid arthritis (RA; 24 seronegative, 39 seropositive) and 15 healthy controls. Patients with SNA and RA had a significant increase of lymphocytes, especially T, T-helper and activated cells. In addition patients with RA had a significant increase of natural killer cells and a lower percentage of alveolar macrophages and T-suppressor cells. Transbronchial biopsy was performed on 9 patients with SNA and on 59 patients with RA. Abnormal histologic features of lung tissue were observed in 4 out of 9 patients with SNA (2 with fibrosis, 1 with follicular lymphoid hyperplasia, 1 with desquamative interstitial pneumonitis). The abnormal lung histology in RA patients was more pronounced, however, the differences between SNA and RA were not significant. The data from BAL and histology suggest, that the pulmonary involvement in SNA and RA is caused by an unspecified immunologic process. PMID- 8235493 TI - Fibromyalgia in the adult Danish population: I. A prevalence study. AB - Epidemiologic studies of fibromyalgia have so far been based on rheumatologic and general practice settings, which are poor proxies for the underlying population. The study is based on a national health interview survey carried out by the Danish Institute for Clinical Epidemiology in 1990/91 on approx. 6000 randomly selected Danish citizens. For this study 1219 subjects from the eastern part of Denmark aged 18 to 79 years were asked about widespread muscle pain. One-hundred and-twenty-three persons fulfilled the screening criteria. Clinical examination could be performed on 65 persons (53%). Eight subjects, all female, met the 1990 American College of Rheumatism criteria for fibromyalgia. Dropouts were regarded as not having fibromyalgia. The prevalence of fibromyalgia in the Danish population between 18 and 79 years of age was found to be a minimum estimate of 0.66% (95% confidence limits 0.28%-1.29%). PMID- 8235494 TI - Fibromyalgia in the adult Danish population: II. A study of clinical features. AB - Clinical characteristics of fibromyalgia have so far been based mainly on patients identified in rheumatologic settings. This paper offers the clinical findings in fibromyalgia based on a national health interview survey, in which 123 persons fulfilled preset criteria for widespread pain. Clinical examination could be performed on 65 subjects (53%) and included physical examination, tender point palpation by two blinded trained physicians, blood sample analysis, measurement of dynamic muscular strength and a detailed self-administered questionnaire. Significantly more subjective swelling, fatigue, headache, difficulty in stair-climbing, and poorer self-evaluated health with more tender points was found. Contrary to that which was expected, fibromyalgia subjects did not suffer from sleep disturbances, irritable bowels or morning stiffness. Our findings indicate that clinical characteristics of fibromyalgia in the general population may differ from those found in rheumatological settings. PMID- 8235495 TI - Ultrasound examination of metacarpophalangeal joints in rheumatoid arthritis. AB - To evaluate the ability of ultrasonography with high frequency transducer (13 MHz) in detecting metacarpophalangeal (MCP) joint abnormalities, 20 patients with rheumatoid arthritis (RA) and a symptomatic involvement of MCP joints were studied. Twenty healthy subjects served as controls. In all RA patients, ultrasonography clearly demonstrated one or more soft tissue and/or bone abnormalities. Sixteen patients (80%) had joint cavity widening because of effusion (1 case), synovial thickening (7 cases), and joint effusion with synovial thickening (8 cases). Loss of definition of the metacarpal articular cartilage was observed in 17 cases (85%). Sixteen (80%) had bone erosions and 9 (45%) showed a definite widening of the flexor tendons sheath. Margin irregularities of the extensor and flexor tendons were respectively visualized in 7 (35%) and in 8 (40%) cases. An extensor tendon rupture was observed in 2 cases (10%). Ultrasound examination by 13-MHz transducer appears to increase the accuracy of MCP joints evaluation in RA with respect to conventional radiography. It can be recommended as the first investigation in the assessment of soft tissue involvement in RA patients. PMID- 8235496 TI - Frequency of amyloidosis in Estonian patients with rheumatoid arthritis. AB - Occurrence of secondary amyloidosis in hospitalized Estonian patients suffering from rheumatoid arthritis was studied using subcutaneous fat aspiration biopsies. Only 47 out of 59 samples included fat, and amyloidosis deposits were found in 11 of these (23%). The high frequency of amyloidosis may be due to selection of the most severe cases in the hospital. PMID- 8235497 TI - Subcutaneous nodular amyloidosis in Sjogren's syndrome. AB - We describe a female patient with primary Sjogren's syndrome (SS) associated with a subcutaneous tumor formed by amyloid surrounded by mature IgG-kappa plasma cells, in the absence of lymphoproliferation or amyloid deposit in other organs. This case represents an unusual presentation of localized amyloidosis, without systemic involvement. Together with other reported cases this suggests that only immunocytic and not reactive systemic amyloidosis develops in chronic SS patients. PMID- 8235498 TI - Monoarticular juvenile chronic arthritis of the shoulder. Report of a case and review of the literature. AB - In 20% of the cases Juvenile Chronic Arthritis (JCA) has a monoarticular onset. Usually the inflammatory process spreads out to other joints with pauciarticular or polyarticular course. Very rarely the disease persists in one joint only and this is in about 70% of the cases, the knee. We describe a case of Monoarticular JCA with isolate and persistent involvement of the shoulder never reported in the literature. The clinical, pathologic features and different diagnoses are discussed. PMID- 8235499 TI - Children with and without disabilities in the Nordic countries. A Nordic project. AB - A socio-pediatric study of children's health and well-being has been carried out, based on questionnaires from 2000 children with long-term illnesses and disabilities and from a representative sample of 10,000 children from the five Nordic countries. The overall results confirm that Nordic children enjoy a high standard of living, and that they are healthy, both physically, mentally and socially. They frequently use the health services but mainly for minor complaints and their hospital stays are short. Disabled children and their families have the same material standard of living and a similar social network as the others. Their leisure time activities, however, are reduced and the children's peer acceptance and self-esteem are lower and they have more psychosomatic symptoms. It is mainly the mothers who have to interrupt their career to take care of the disabled children. Most families, both disabled and others, are quite satisfied with the children's medical care, especially where continuity and specialist services are provided. PMID- 8235500 TI - Morbidity and mortality in Estonia, Latvia and Lithuania in the 1980's. AB - The purpose of the present study was to examine the general morbidity and mortality rates in the three Baltic republics--Estonia, Latvia and Lithuania during a decade before the collapse of the Soviet Union. Official statistical data were used to compare morbidity and mortality rates. A method of standardization and life table functions were employed. Soviet morbidity statistics were predominantly descriptive, and based mainly on crude rates registered cases of illness during a year per 100,000 population. The death rates during the Soviet period are a better indicator of the health of the populations than more specific health indicators. A general deterioration of the ecological, social and economic situation in the Baltic republics at the end of the 1980s and early 1990s may have resulted in the mortality rising and aborted any trends to increasing life expectancy in the next few years thus requiring more radical reforms in health and social policy. The study demonstrates a noticeable difference in mortality rates between demographic and socio-economic groups in the Baltic states. PMID- 8235501 TI - Self-reported health and function status of elderly Turkish immigrants in Copenhagen, Denmark. AB - This study analyses the self-reported health and functional status of elderly Turkish immigrants in Copenhagen. One hundred and sixteen Turkish immigrants aged 55 and over were interviewed at home about their health problems and their ability to perform a number of daily activities. Only one-third of the respondents reported having no chronic illness, while chronic arthritis was the one mentioned most frequently (41%). Almost 2/3 of the respondents reported having pains in their locomotor systems. Women are overrepresented, but the differences are not statistically significant. As regards the 11 daily activities in the questionnaire, only 39 of the respondents reported being able to perform all of them without difficulty. The article suggests some possible explanations for the results and suggests future areas for research. PMID- 8235502 TI - Precursors of repeated short-term sick-leave: an empirical review of some background, job and well-being characteristics. AB - The aim of the study was to search for bivariate associations between on the one hand samples of background, job-related and well-being characteristics, and on the other hand longitudinally recorded sick-leave in individuals with repeated short-term sick-leave. Out of altogether 123 subjects included, 113 self-rated their well-being at the onset of, and 88 also after, a supportive programme. The well-being rating covered experience of treatment by other people, reservation, loneliness, inferiority, tension, vulnerability, guilt, security and indolence. Socio-demographic and job-related characteristics and attitudes were registered and declared at the initial contact. Correlations between these separate sets of independent variables, and sick-leave occasions and days in each of the two years following the contact were computed. There was an almost complete lack of covariation between background variables and sick-leave. Neither did job-related characteristics correlate. Only an expressed wish for a job change was vaguely associated with more absence days, but not with more spells. In striking contrast, a convincing consistent longitudinal pattern between self-rated well being and sick-leave behaviour emerged. The poorer the well-being, the greater the subsequent sick-leave. Yet, a major part of the sick-leave variance remained unexplained. Accordingly, other etiological factors may still interact with well being in the comprehensive setting behind the sick-leave behaviour.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235503 TI - Working at the loom and musculoskeletal disorders in a female population of Crete, Greece. AB - The influence of the working at the loom on the appearance of musculoskeletal disorders has been studied in the adult female population of a highland community of Crete, in which home-based handicraft has an important place in the economic activity. Information about symptoms and clinical signs related to the musculoskeletal system, as well as about socioeconomic characteristics was collected by means of a personal interview and a medical examination. Regression analysis shows that there is a statistically significant association between the intensity of symptoms from the upper extremities and the shoulder-neck region and the number of years of working at the loom, independently from other studied variables, such as BMI, age and number of children per woman. PMID- 8235504 TI - Indicators of "avoidable" mortality in health administrative areas in Sweden 1974 1985. AB - When comparing health administrative areas in Sweden the variation in death rates for 13 suggested indicators of avoidable mortality was analysed for the time periods 1974-79 and 1980-85. For most indicators the variation was significant. The level of systematic (nonrandom) variance differed between the sexes for certain conditions. According to the theory of mass-significance it would be appropriate to use a sharpened significance test of high SMRs. For certain health policy indicators it was possible to detect areas accounting for nearly all the deviation above the national standard using a sharpened test (p < 0.001). For most medical care indicators, however, the high death rates had a lower level of significance. When sensitivity is given priority an ordinary test (p < 0.05) should be preferred. This means, however, dealing with potential false positive warning signals. Epidemiological surveillance of indicators of avoidable mortality should be the starting point for in-depth studies. PMID- 8235505 TI - Modernization and smoking--regional differences in adolescent smoking in Finland in 1977-1987. AB - The purpose of the study was to test the applicability of the modernization framework to changes and regional diversity in adolescent smoking. A biannual cross-sectional survey data from the Adolescent Health and Lifestyle Survey was used. A national sample of 14, 16- and 18-year-old Finns answered a mailed questionnaire every other year from 1977. The 1977, 1979 and 1981 data were pooled as were data from 1983 and 1985 (N = 8505, 9777). The 1987 sample was weighted in accordance with the province size (N = 11,374). Modernization trends found in the literature were apparent among adolescent smokers between 1977 and 1987. The use of hand-rolled cigarettes nearly stopped when the use of manufactured cigarettes increased. The use of low-tar cigarettes increased and that of other tobacco types decreased. Regional differences in tobacco types were associated with the structural modernization level of the regions. Snuff use was most prevalent in the most modern and the most traditional regions. The modernization framework proved useful in explaining changes in smoking. PMID- 8235506 TI - Predictors of smoking cessation in pregnancy. AB - Smoking cessation is difficult to obtain, even among pregnant women who should be highly motivated to quit. Identification of predictors of unsuccessful smoking cessation may be of help in identifying pregnant smokers who need special attention in order to reduce or eliminate the intrauterine tobacco smoke exposure. The study was done in two consecutive samples of pregnant women in Odense and Aalborg, Denmark. Altogether 13,815 pregnant women participated in the study by reporting detailed information on their lifestyle during pregnancy and before pregnancy. Recruitment of pregnant women started in 1984 and ended in 1987. The study is based upon self-administered questionnaires filled out in the 36th week of gestation. Only those who were smokers prior to the pregnancy are included in the study (5,724). The study showed that the strongest predictors of unsuccessful smoking cessation were: the level of smoking prior to pregnancy, coffee consumption, and the partners smoking habits. Parity, school education, and social status also played a role but were less strongly associated with continuous smoking in pregnancy. PMID- 8235507 TI - Students opinions about sex education in a Swedish school region. AB - Sex education has been a requirement in Swedish schools for the past 30 years. The purpose of the study was to describe the opinions of 17- and 18-year-old Swedish students at the upper secondary school regarding the sex education they received at school as well as their knowledge about the anatomy and physiology of reproduction. A questionnaire concerning education about sex and relationships was answered by 421 students. The results of questions regarding human reproduction illustrated that the students have insufficient knowledge about men's and women's reproductive organs. Two-thirds of the students who had received sex education thought the instructions were insufficient. The students wanted to talk more about positive and negative feelings within a relationship and feelings concerning sexually transmitted diseases and abortion. In summary, the results indicated that the content of sex education should focus on women's reproductive organs and the feelings and relationships between men and women. The type of instructions and how instruction time is utilized should be examined closely. PMID- 8235508 TI - A study of hospital admissions over time, using longitudinal latent structure analysis. AB - The aim was to study patterns of utilization of non-psychiatric admissions over time and factors affecting the utilization. The study cohort includes all individuals born 1934-66, living in one of two Danish municipalities and admitted to a non-psychiatric department at least once in 1977 (n = 2,686). The hospitalizations of the cohort were followed during a 5-year period by means of the Danish National Patient Register. The data were analysed using a longitudinal latent class (LC) model and a longitudinal latent Markov (LM) model. The LC model suggests that among the cases in the cohort there were 4 variants of utilization patterns. The LM model adequately described the sample in only 3 variants or classes. These classes may be interpreted as a small group of "chronically ill" individuals (1.9% of the cohort), a major group of "healthy" individuals, with no, or only a single, random re-admission during the follow-up period (74.4% of the sample), and finally an intermediate group of "high utilizers" (23.7% of the sample). This "chronicity" variable was markedly associated with mental illness, multiple discharge diagnoses from non-psychiatric departments and total utilization of hospitalizations during the follow-up period. Conversely, gender, age and days in hospital per admission were without importance. The study implies that the analysis of patterns of hospital admissions over time can yield important insight into health service utilization and that longitudinal latent structure analyses are powerful statistical tools in this aspect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235509 TI - "Project lifestyle" and new forms of living for the mentally retarded in the local community. AB - Since 1890, representatives of the Ministries and Departments of Social Security in Norway, Sweden, Finland, Iceland and Denmark have been working to create a foundation and a framework for a common Scandinavian research and development project concerning the quality of life of the mentally retarded. In 1992, with financial support from The Nordic Council of Ministers, the exchange of information and the building up of a network was begun, involving those taking part in some 50 development projects in Scandinavian countries. The project has focussed on three themes central to efforts made on behalf of and with the cooperation of mentally retarded people in these countries: the mentally handicapped person in the local community, independence and the quality of life, and communication. The aim of the project is to establish a Scandinavian network of key persons in the field, to systematize and propagate promising developments and to cast light on differences and similarities between various Scandinavian models for research and development related to the mentally retarded. PMID- 8235510 TI - Observations on the dose-response curve for arsenic exposure and lung cancer. AB - Occupational studies in three countries have related quantitative estimates of arsenic exposure to lung cancer risks. Mine exposures in China appear to incur a higher relative risk than arsenic exposures elsewhere. All of the studies with quantitative data are consistent with a supralinear dose-response relationship. Two studies are also consistent with a linear relationship over an elevated background risk of lung cancer among arsenic-exposed workers. Neither toxicokinetic mechanisms nor confounding from age, smoking, or other workplace carcinogens that differ by exposure level appear likely to explain this curvilinearity. Plausible explanations include (i) synergism (with smoking) which varies in magnitude according to the level of arsenic exposure, (ii) long-term survivorship in higher exposure jobs among the healthier, less susceptible individuals, (iii) exposure estimate errors that were more prominent at higher exposure levels as a result of past industrial hygiene sampling or worker protection practices. PMID- 8235511 TI - Pesticide poisonings in Costa Rica. AB - A descriptive epidemiologic study, conducted in Costa Rica, investigated the incidence of pesticide poisonings with special attention to agricultural workers and occupational exposure. Information from three national registers (occupational accident and disease reports, hospitalizations, and deaths) were used. During 1986, 1800 occupational accidents caused by pesticides were reported; between 1980 and 1986 altogether 3330 persons were hospitalized and 429 died. Cholinesterase inhibitors caused 71% of the reported occupational accidents, 63% of the hospitalizations, and 36% of the deaths. Paraquat caused 21% of the occupational accidents, 24% of the hospitalizations, and 60% of the deaths. Hospitalizations and deaths were 13 and 11 times, respectively, more frequent among agricultural workers than among the rest of the population. High risk groups for occupational poisonings included agricultural workers aged 15-29 years, female workers, and banana plantation workers. The yearly incidence of symptomatic occupational pesticide poisonings among agricultural workers was estimated at 4.5%. PMID- 8235512 TI - Solvent exposure, alcohol consumption and liver injury in workers manufacturing paint. AB - Liver enzyme activity was examined in 89 South African paint makers currently exposed to a mixture of organic solvents at fairly low levels. However, the duration of exposure was substantial for many. Fifty-eight workers (65%) had at least one enzyme value above the upper reference limit. Gamma glutamyl transferase (gamma GT) activity was elevated in 46% of the workers and aspartate aminotransferase (ASAT) in 52%. In a comparison between workers with high and low solvent exposure, the gamma GT and ASAT values were higher in the most exposed group (eg, gamma GT mean 108 versus 69 U.l-1, P > 0.05). Adjustment for confounding by alcohol consumption and body mass index eliminated the differences due to exposure. It was concluded that the measures of liver injury used did not demonstrate solvent-induced hepatic damage but that excessive alcohol consumption was an important factor. PMID- 8235513 TI - Effects of occupational exposure to organic solvents and noise on hearing. AB - This study explored the effects of occupational exposure to solvents and noise on hearing. Interviews and hearing tests were conducted for printing and paint manufacturing workers. The experimental groups included unexposed (N = 50) workers and workers exposed to noise (N = 50), noise and toluene (N = 51), or an organic solvent mixture (N = 39). The risk of hearing loss was greater for the exposed groups than for the unexposed group. The adjusted relative risk estimates were four times greater [95% confidence interval (95% CI) 1.4-12.2] for the noise group, 11 times greater (95% CI 4.1-28.9) for the noise and toluene group, and five times greater (95% CI 1.4-17.5) for the solvent-mixture group. The findings suggest that exposure to the studied solvents had a toxic effect on the auditory system and that an interaction between noise and toluene took place. The audiological results of the noise and toluene group suggest a central auditory pathway involvement in the hearing losses observed. PMID- 8235514 TI - Monitoring occupational exposure to styrene from hemoglobin adducts and metabolites in blood. AB - Monitoring occupational exposure to styrene was achieved through quantification of adducts of styrene 7,8-oxide to N-terminal valine in hemoglobin (Hb) on the basis of the enrichment of adducted globin chains by ion-exchange chromatography and gas chromatographic-mass spectrometric analysis by the use of the N-alkyl Edman method. Application to blood samples from reinforced plastics workers exposed to styrene and from referents showed Hb adduct levels correlating with the blood styrene glycol and urinary mandelic acid concentrations. The blood styrene glycol and styrene 7,8-oxide levels of the exposed workers averaged 2.5 mumol.l-1 (17 subjects) and 0.09 mumol.l-1 (7 subjects), respectively. The blood styrene glycol and urinary mandelic acid content (mean 9.5 mmol.l-1, 17 subjects) suggested a styrene concentration of about 300 mg.m-3 (75 ppm) in the workplace air. The Hb adduct levels were low (mean 28 pmol.g-1), indicating rapid detoxification of styrene 7,8-oxide in humans. PMID- 8235515 TI - Neurobehavioral functions among workers exposed to manganese ore. AB - The Neurobehavioral Core Test Battery of the World Health Organization was performed on 17 baggers (mean age 36.6 years) in a manganese ore milling plant and 17 referents (mean age 35.7 years). The baggers had a mean exposure period of 7.4 (range 1-14) years with a mean blood manganese concentration of 25.3 (range 15-92.5) micrograms.l-1 [0.46 (0.27-1.68) mumol.l-1]. Of the 37 symptoms related to the nervous system, 20 were more frequently reported by the exposed workers. No significant differences were noted between the nerve conduction of the median and ulnar nerves of the dominant forearm of the two groups. The exposed workers had significantly poorer motor speed, visual scanning, visuomotor coordination, visuomotor and response speed, and visuomotor coordination and steadiness. A clinical examination did not reveal any abnormality among the two groups. Hence a neurobehavioral test battery may be a more sensitive method than a clinical examination in detecting early changes in motor function among manganese-exposed workers. PMID- 8235516 TI - Digital arterial responsiveness to cold in healthy men, vibration white finger and primary Raynaud's phenomenon. AB - Finger systolic pressure (FSP) was measured during a standardized cold test in 291 healthy male subjects divided into five age groups. The age groups showed no difference in the change in FSP (FSP%) after cold provocation at 15 and 10 degrees C. In the entire population, the lower normal limits of FSP% were estimated as 76% at 15 degrees C and 63% at 10 degrees C. When a discriminating threshold of FSP%10 degrees < 60% was applied to the results of the cold test of 31 referents, 65 chain saw workers with or without vibration-induced white finger (VWF), and 20 male patients with primary Raynaud's phenomenon (PRP), the sensitivity of the test to detect digital vasospasm was 84% for VWF and 95% for PRP. In the chain-saw worker group, the positive and negative predictive values of the cold test were 94%. Therefore the measurement of FSP during cold provocation can be considered a useful laboratory test to confirm Raynaud's symptoms objectively in both groups and individuals. PMID- 8235517 TI - Development of trapezius myalgia among female workers performing light manual work. AB - The aim of this prospective study was to detect a possible development of trapezius myalgia likely to be work-related in initially healthy female packers. Symptoms were recorded on a five-step intensity scale in a daily diary and in an interview at ten-week intervals. All of the subjects were interviewed concerning assumed risk factors during work and leisure time, and a clinical examination was performed. Within their first year of employment, 17 of 30 subjects developed work-related trapezius myalgia of sufficient intensity to be classified as patients. The median time before a clinically confirmed patient status was 23 (range 6-51) weeks. The symptoms showed a fluctuating pattern, decreasing on the weekends and during some holidays. In conclusion, likely work-related trapezius myalgia of a nonchronic character showed a high one-year cumulative incidence at a plant with repetitive light work. PMID- 8235518 TI - Electromyographic evaluation of muscular work pattern as a predictor of trapezius myalgia. AB - Electromyographic (EMG) measurements and interviews concerning muscular pain and disability were performed prospectively every 10th week on 30 healthy new female employees of a chocolate manufacturing plant. The static muscle activity and rate of short unconscious interruptions in EMG activity (EMG gaps) of the trapezius muscle during repetitive work tasks were evaluated as possible risk factors for patient status with trapezius myalgia. At the start of employment, the static and median contraction levels were significantly higher in future patients than in the rest of the subjects (nonpatients). In subsequent recordings these values were reduced to the level of the nonpatients. Throughout the study, the future patients had a lower frequency of EMG gaps than the nonpatients. A regression analysis showed a significant value for a low rate of EMG gaps to predict future patient status. PMID- 8235520 TI - [New guidelines and another layout]. PMID- 8235519 TI - Meeting of the IARC working group on occupational exposures of hairdressers and barbers and on some colorings and dyestuffs. PMID- 8235521 TI - Overdenture abutments and reduced periodontium in elderly patients. A retrospective study. AB - Among elderly patients attending a private office, a large number was seen having only few remaining teeth with advanced reduction of periodontal support. The long term effect of overdenture rehabilitation within a mean observation time of 5.9 years (2 to 10 years) could be assessed in 109 patients (average age 74.1 years) wearing a total of 125 overdentures. After completion of treatment, 76% had regularly followed the maintenance care program. The degree of bone loss of the selected abutment teeth was assessed by means of radiographs. 56% showed a highly reduced periodontal support with bone loss of more than 50% of the root length. No, or moderate loss of periodontal support was found in only 9.5%. The overdentures were initially retained by 359 cast gold copings. Biologic and technical failures were registered for 54 copings and examined with regard to the reduction of periodontal support of the abutment teeth. Repair and treatment were prescribed for 30 copings, including guided tissue regeneration for 6 abutments. Twenty-four abutment roots with copings were extracted. Although most biologic failures were encountered in abutment teeth with severe reduction of the periodontium, the failure rate was not statistically higher for these abutments. PMID- 8235522 TI - [A comparison of 3 laser systems for dental enamel ablation]. AB - Three different laser systems were compared according to their applicability in dentistry: a Ho:YAG laser (lambda = 2, 12 microns, pulse duration = 250 microseconds and 4 microseconds, pulse energy up to 300 mJ), an Er:YAG laser (lambda = 2,94 microns, pulse duration = 60-100 microseconds, pulse energy up to 100 mJ) and an in dentistry so far unknown picosecond Nd:YLF laser (lambda = 1,053 microns, pulse duration = 30 ps, pulse energy up to 1 mJ). The morphological changes of the enamel due to laser radiation were studied using light and scanning electron microscopy. Possible enamel fractures were visualized with dye penetration tests. Thermally caused, about 3 mm long enamel fractures were only observed using the Ho:YAG laser. Smaller fractures, up to 300 microns, were generated with the Ho:YAG and the Er:YAG laser. The Nd:YLF laser produced well defined square cavities with a high precision. No enamel fractures could be observed. The dye penetration in the cavity walls was less than 20 microns. PMID- 8235523 TI - Effects of veneering and glazing on the strength of heat-pressed ceramics. AB - A newly developed press-type all-ceramic crown system, the IPS-Empress system (Ivoclar), has recently been introduced. Two methods may be used to obtain the desired shade: surface staining and glazing; veneer technique. The purpose of this study was to determine whether these two methods affected flexure strength of Empress glass ceramic. Eight groups of test bars were pressed. In groups 1 and 2, one surface was stained and glazed. The bars were placed face down (1) or up (2) for testing. For comparison, group 3 was heat-treated only (simulating stain and glaze firing). In groups 4-7, one surface of the bars was either veneered with porcelain on the bottom (4) or top surface (5) and then subsequently glazed (6 and 7). Group 8 was just heat-treated (simulating veneer and glaze firings). The results showed that there were no significant differences in strength between groups 2, 5 and 7 compared to the reference groups 3 and 8 (159 +/- 28 and 175 +/ 32 MPa, respectively), indicating that, from the mechanical point of view, the two surface techniques can be equally used on Empress ceramic. If the porcelain veneer supported the ceramic (4), the strength was significantly decreased. The highest mean value was obtained in group 1 (220 +/- 34 MPa). PMID- 8235524 TI - [The strength of ceramic and polymer after CAD/CIM treatment and in a bond with dentin]. AB - The flexural strength of bar samples (2 x 4 x 12 mm) of Vita Mark II, Dicor MGC Dark & Light and CR-Polymer was examined after machining with the Cerec E-Drive unit using grinding wheels coated with coarse (126 microns) and fine (46 microns) diamonds. The flexural strengths of both Vita MK II (98.5-112.4 MPa) and Dicor MGC Dark (135.1-205.3 MPa) were significantly (p < 0.01) different after machining with the coarse and fine discs respectively. The flexural strengths of both Dicor-MGC Light (192.6-220.1 MPa) and of CR-Polymer (103.9-104.3 MPa) were not significantly changed by machining with coarse or fine diamonds, respectively. Bonded sandwich samples (2 x 4 x 12 mm) were made from parts (1.45 x 4 x 12 mm) of Vita MK II, MGC D&L and CR-Polymer and of parts (0.45 x 4 x 12 mm) of bovine dentin using a dentin adhesive and a 100 microns layer of fine hybrid composite resin. The flexural strengths of the samples machined from porcelain and glass ceramic (2 x 4 x 12 mm) and the sandwich samples composed of ceramic (1.45 x 4 x 12 mm) and bovine dentin (0.45 x 4 x 12 mm), respectively, did not differ significantly (p > 0.001): Vita MK II 106.4 +/- 12.2/98.5 +/- 25.8 MPa. Dicor MGC Dark 195.6 +/- 34.6/145.6 +/- 31.2 MPa. Dicor MGC Light 204.1 +/- 29.1/171.5 +/- 18.2 MPa.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235525 TI - [Gingival recession. A review of its prevalence, etiology and classification]. PMID- 8235526 TI - [Corrective therapy in gingival recession. Langer's subepithelial connective tissue graft and Holbrook-Ochsenbein's free mucosal transplant--methods and case examples]. PMID- 8235527 TI - [New methods for the covering of exposed necks of teeth. A modification of the connective tissue transplant]. PMID- 8235528 TI - [Fluorides and fluoridation: a declaration of principles of the FDI. Federation Dentaire Internationale]. PMID- 8235529 TI - [The HIV test in Switzerland--an epidemiological position determination?]. PMID- 8235530 TI - [Dentistry and the environment ... D-day soon!]. PMID- 8235531 TI - [The evaluation of the cytotoxicity of 2 dental adhesives using human pulp cells in culture]. AB - This in vitro study was designed to ascertain the cytotoxicity of two new dentin systems, Scotchbond MP (3M) and A.R.T. Bond (Coltene), on cultured cell monolayers prepared from fresh explants of human pulp tissue and placed in contact with the materials according to two methods. In a direct method, the two components (primer, adhesive) of each bonding system were placed directly on the cell layers. In an indirect method the bonding agents were placed on dentin slices of 0.3 and 1.0 mm thickness, that were interposed between agents and cells. After 8 days the cytotoxicity was quantitatively assessed by counting of the remaining living cells. The results indicate that the different primer solutions, when placed in direct contact with the cells, are more cytotoxic than the adhesive resins. In indirect contact through dentin, both bonding systems exhibited a similar cytotoxicity. The cytotoxicity appeared to be considerably diminished with a thick dentin slice interposed. These data confirm the role of dentin as a diffusion and buffer membrane for bonding materials. This kind of experiment could in the future improve the correlation between in vitro and in vivo results. PMID- 8235532 TI - [Microscopic studies of the structures of different dental floss types]. AB - 26 different dental flosses, made from silk, polyamide (Nylon) or Teflon, were analyzed with respect to their surface structure and their cross-section. Principally, two different types of manufacturing could be distinguished. The dental flosses either consisted of a large number of individual fibers, or a single, falted membrane. Even flosses that were manufactured in the same manner revealed considerable differences. Apart from the chemical composition, the structure of dental flosses is decisive for their use and this is why an exact classification would be desirable. In this study the requirements for classifying dental flosses are provided. PMID- 8235533 TI - [The effect of the aeration status on the corrosion behavior of Ni-Cr alloys]. AB - Using in vitro tests, the influence of aeration cells that develop in narrow crevices of dental restorations, on the corrosion behaviour of two Ni-Cr alloys (Wiron 88 and Rexillium III) was investigated. The results revealed a 1.6-fold higher ion release in the deaerated part of the electrolyte compared with the aerated one, whereas the ion release was 18-fold higher with Rexillium III. The total analyzed substance loss was 11.1 (S.D. 5.7) and 181.9 (S.D. 48.0) micrograms/cm2, respectively. PMID- 8235534 TI - [The germ colonization of the oral cavity in small children with the nursing bottle syndrome]. AB - 46 infants, aged 23 to 77 months, presenting with the nursing bottle syndrome, were examined clinically [dmf(t)] and microbiologically. Saliva and carious tooth substance was analysed for the presence of Streptococcus mutans, Lactobacilli and Candida albicans, using in part commercial tests. For comparative reasons, a group of caries-free children, aged 37 to 77 months, was examined as well. Increased numbers of specific caries bacteria (mutans streptococci and lactobacilli) and of Candida albicans were found in saliva and in the carious material of children with diseased teeth. The results confirm that excessive drinking from nursing bottles over a period longer than the first 12 months of life does not only lead to problematic tooth decay and its consequences but also to pathological changes in the entire oral flora. The invasion of candida forces us to reconsider the development of caries in infants. PMID- 8235535 TI - [The hygienically correct practice concept (II). Passive protective measures (inoculations, work protection, work clothes, gloves, mouth protection, protective glasses). The Commission for Practice Hygiene of the Societe Suisse d'Odonto-Stomatologie]. PMID- 8235536 TI - [Practical dentistry in the 18th century. Historical grave findings from Saint Hippolyte, Le Grand-Sacconnex GE]. PMID- 8235537 TI - [Implants in the atrophic jaw. A report on the technical meeting of the Swiss Society for Implantology (SSI) of 7 June 1993 in Basel]. PMID- 8235538 TI - [Bone regeneration in theory and practice. A report on the continuing education course of 7 June 1993 at the Basel Exhibits and Convention Center]. PMID- 8235539 TI - [Amalgam separators: the current status]. PMID- 8235540 TI - [Swiss hygienists: already 20 years!. Interview by Francois Reymond and Kurt Venner]. PMID- 8235541 TI - [Use in the cat of the "small external fixator for hand and forearm" of the Association for Osteosynthesis Inquiry. I. Introduction]. AB - In the first part general principles and indications for external fixator use in cats for treatment of fractures and luxations are described. This study considers only one of many external fixator systems. The results of the clinical use of this system in cats are described in the second part of this investigation. Technical aspects, post operative care, implants removal, possible complications and their therapy are discussed. PMID- 8235542 TI - [The effect of pregnancy and lactation in sows on the pharmacokinetics of the gyrase inhibitor marbofloxacin]. AB - Six pregnant sows were treated in early pregnancy, late pregnancy and during lactation. Four empty sows served as control. Marbofloxacin was administered (2 mg/kg body weight) intravenously and orally. The active drug concentration in the plasma was quantitated by use of high performance liquid chromatography (HPLC) and microbiological assay. Pharmacokinetic parameters were calculated by use of a noncompartment model. In lactating animals, the concentrations in milk were also determined by HPLC. Mean elimination half-life of the drug after oral administration was significantly shorter in lactating sows (5.74 h) than that of the early pregnancy group (10.09 h). Total body clearance was highest in the lactating sows (3.27 ml/minute.kg body weight). The volume of distribution was large in all physiological states studied indicating good tissue penetration. Bioavailability was about 80% in pregnant and lactating sows. Antimicrobial secretion in milk contributed greatly to marbofloxacin elimination. These results indicate an important influence of lactation on marbofloxacin pharmacokinetics in sows. Therefore, in such cases, the antibiotic dose should be increased during lactation. PMID- 8235543 TI - [Histological and histochemical studies of the innervation of the teat and the skin of the bovine udder]. AB - The nervous supply of the teat and udder skin of the bovine was examined by means of the AChE-reaction. In comparison to the silver impregnation method, a rich subepidermal fibre-network with intra-epithelial nerve endings and, especially, a strongly developed nerve fibre meshwork associated with muscle fibres was seen. PMID- 8235544 TI - [The concentration of ascorbic acid in the liver, the cerebrum, the kidney, the adrenal glands and the musculus longissimus dorsi of dog fetuses, dogs and cats of different ages]. AB - In dog fetuses aged of 45 and 55 days as well as in dogs and cats of different ages the body weight and the concentration of ascorbic acid in different tissues were analysed. In the liver, the highest concentration was determined in the dogs aged 28 days (432 +/- 71 micrograms/g wet weight) and in the cats aged of 42 days (474 +/- 85 micrograms/g). In the cerebrum the highest concentration was found in dog fetuses aged 55 days (821 +/- 75 micrograms/g) and in cats aged 1 to 3 days (888 +/- 56 micrograms/g). In the kidneys and in the M. longissimus dorsi, the concentration was also in part age-dependent. The concentration in the adrenals of dog fetuses was lower than that after birth: the highest concentration was analysed in dogs aged 28 days (1780 +/- 340 micrograms/g) and in cats aged 6 months (1497 +/- 203 micrograms/g). The significance of ascorbic acid for the development of the tissues of carnivores is discussed and compared with the concentration in other species. PMID- 8235545 TI - [What is your diagnosis? Sterile, suppurating meningitis]. PMID- 8235546 TI - Toward the design of a new bowel care chair for the spinal cord injured: a pilot study. AB - Bowel care is a critical aspect of daily living for person with disabilities; ineffective bowel care can lead to severe and costly complications. Staff, patients, and caregivers have often found existing bowel care chairs to be inadequate. These chairs are frequently unsafe, inconvenient, and ineffective for showering and bowel elimination. Because bowel care procedures can be lengthy, proper seating posture and comfort is necessary to prevent pressure ulcers. Patient falls is another common problem, occurring during transfer, transport, or during actual bowel care/shower procedures. There are over 200,000 persons with spinal cord injuries (SCI) in the United States today. The majority of these patients have neurogenic bowels, requiring bowel care an average of three times a week. Each bowel care procedure takes 30 minutes to 3 hours to complete. Advances in bowel care chair design would significantly impact on quality of life, self esteem, and physical well-being of persons with SCI. Enhanced design would also improve the safety, effectiveness, and efficiency of nurses and caregivers who perform bowel care procedures. An interdisciplinary research team, comprised of an industrial designer, a nurse, two physicians, and a human factors psychologist collaborated to address this important problem. Three bowel care chairs, commonly used in SCI, were evaluated using a combination of videotaping, still photography, and questionnaires. Based on this data, performance criteria for the design of an optimal bowel care chair design were developed. PMID- 8235547 TI - The prevention of violent spinal cord injuries. AB - Interpersonal violence has been defined by the USPHS as a national critical health care problem. Of the 10,000 to 12,000 new spinal cord injuries in the United States each year, about a 1,000 will be caused by gunshot wounds. Violence is identified as part of our heritage, inherent in our entertainment and, sadly, part of our everyday life. Community response to these intentional injuries has indicated they will no longer be ignored. Conventional prevention efforts are often not relevant. Violence prevention efforts require the focus to be upon the critical precipitating factors. The development of school, community and youth based violence-specific injury prevention programs are essential. PMID- 8235548 TI - Advancement in computer technology: a tool to identify seating problems and prevent pressure ulcers. PMID- 8235549 TI - Hard times. Occupational injuries among children are increasing. PMID- 8235550 TI - Evolutionarily mobile modules in proteins. PMID- 8235551 TI - Simulating brain damage. PMID- 8235552 TI - More fun than a root canal. PMID- 8235553 TI - Were four corners victims biowar casualties? PMID- 8235554 TI - Unraveling Alzheimer's. A major cause of the disease yields to researchers. PMID- 8235555 TI - Chemical signaling in the brain. PMID- 8235556 TI - Ancient DNA. PMID- 8235557 TI - Prevention of coronary disease in women. PMID- 8235558 TI - The Scottish Renal Registry. PMID- 8235559 TI - Home parenteral nutrition in Tayside 1980-1992. AB - The experience of Home Parenteral Nutrition (HPN) in Tayside over the last 13 years has been reviewed with particular reference to the indications for, complications of, and quality of life whilst receiving HPN. The observed complication rate has been compared with that of the UK experience, and the observed complication rate for the first seven years of HPN experience has been compared with the subsequent six years. The Tayside experience equates broadly with that of the UK, although there was a different pattern of complications. Increasing experience with HPN has resulted in a fall in HPN related complications to 1 per 3.4 treatment years in Tayside, a rate as low as that of the most experienced HPN centre in the UK. PMID- 8235560 TI - Dilated cardiomyopathy in haemophiliacs infected with the human immunodeficiency virus. AB - Infection with the human immunodeficiency virus (HIV) can result in several cardiac abnormalities including dilated cardiomyopathy. These phenomena have been described in people contracting the virus through sexual intercourse, injection drug use and by vertical transmission. We have identified recently two Scottish haemophiliacs who have developed dilated cardiomyopathy in the context of HIV infection acquired through treatment with contaminated factor VIII. The significance of this finding is discussed. PMID- 8235561 TI - Protein C and S deficiency causing childhood stroke. AB - The annual incidence of cerebrovascular disease in children is 2.5/100,000 and cerebral infarction is being increasingly recognised in neonates. Deficiency of proteins C and S and their roles in thrombosis have only recently been recognised. Immunologic and functional assays of these proteins now make it possible to determine whether deficiency of them is associated in any particular case of childhood cerebrovascular accident (CVA). We describe two patients, both presenting with stroke in childhood, who were found to be deficient, one in protein C and one in protein S. PMID- 8235562 TI - An audit of hospital discharge letters in patients admitted with acute asthma. Tayside Asthma Audit Group. AB - The management of patients following discharge with acute asthma is dependent on effective communication with general practitioners. A retrospective audit was therefore performed on copies kept of 81 typed discharge letters with a diagnosis of acute asthma between March and October 1991. A subset of 42 interim (handwritten) letters were also audited where case notes could be retrieved. Details on clinic follow up were also obtained from the case notes (n = 42). In the typed letter documentation of severity and treatment of the acute attack was accurate in most cases. Information concerning a precipitating factor was provided in 54% of cases and a smoking history in 57%. Deficiencies were found in specifying inhaler delivery devices (40% recorded), and whether inhaler technique had been formally assessed whilst in hospital (17%). Drug prescribing on discharge was as follows: oral steroid (69%), inhaled steroids (77%), inhaled B2 agonists (92%), theophylline slow release (38%), salbutamol controlled release (20%), and antibiotics (30%). The implementation of a self-management plan and domiciliary peak flow was mentioned in 66% of the letters. The interim letter was generally poor in particular for mention if discharge peak flow (2%), clinic follow-up (64%) and prednisolone regime (61%). Mean +/- s.d. time for clinic follow-up (n = 42) was 4.7 +/- 1.7 weeks (range 1-13 weeks) with 24% non attendance. Thus, improvements in discharge letters are clearly required for optimum continuity of care in the community. PMID- 8235563 TI - Fifty years after: some experiences of the medical care of prisoners of war. AB - Mention is made of Italian prisoners-of-war treated in Catterick military hospital but in October 1943 Italy joined the Allied side. An account is given of the medical condition of 78 Japanese POWs and of 228 members of the Japanese Indian Forces (JIFs) in a transit hospital in eastern India (now Bangladesh). Tropical sprue appeared to occur in an epidemic form each June and July in Burma and eastern India, but did not seem to have affected the Japanese or JIFs. A brief account is given of the condition of 180 European or Australian service men who had survived harsh conditions in Japanese POW camps until liberation in September 1945, then being admitted to a military hospital in Rangoon en route for home. Some had suffered from blindness as a result of malnutrition, the precise reason being uncertain, but there was some improvement after the injection of crude liver extract. PMID- 8235564 TI - The field management of hypothermic casualties arising from Scottish mountain accidents. AB - The purpose of this review is to examine the current literature on hypothermia as it applies to exposure hypothermia of the type encountered in the mountains of Scotland. A brief overview of the problem, illustrated by a case example, is given. The current literature is then explored and then a set of guidelines for mountain rescue personnel is proposed. It is hoped that this article will stimulate debate on the subject to facilitate the development and dissemination of any such guidelines. PMID- 8235565 TI - Health care reform. PMID- 8235566 TI - Non-immune hydrops associated with congenital herpes simplex infection. AB - Non-immune hydrops fetalis is a rare pregnancy complication, which can be caused by a myriad of conditions. Etiologies include metabolic or chromosomal disorders, fetal structural anomalies, and fetal infections. We present a case of non-immune hydrops caused by an intra-uterine infection with Type 1 Herpes Simplex Virus. Evaluation of the fetus with non-immune hydrops should include an amniocentesis for viral culture of the amniotic fluid. PMID- 8235567 TI - Is there an optimal modern method for the diagnosis of breast carcinoma? PMID- 8235568 TI - Thoughtful restraint. PMID- 8235569 TI - Statewide lung cancer study. South Dakota Tumor Registrars' Association. PMID- 8235570 TI - Drug therapy for hyperlipidemia. PMID- 8235571 TI - Memory, transient global amnesia, and history taking. PMID- 8235572 TI - Patient counseling: there is a need. PMID- 8235573 TI - Urinary mercury level in Japanese school children: influence of dental amalgam fillings and fish eating habits. AB - The influence of dental amalgam fillings and fish eating frequency on the urinary mercury (Hg) level was evaluated in 1642 children (ages: 3-18) living in Tokyo. Geometric mean of the urinary Hg level was 1.9 microgram Hg/l and 1.9 micrograms Hg/g creatinine (Cr) for boys and 2.1 micrograms Hg/l and 2.0 micrograms Hg/g Cr for girls. Multiple regression analysis was conducted using: (1) urinary Cr concentration (only in the case of urinary Hg expressed as microgram Hg/l); (2) age; (3) gender; (4) number of dental amalgam fillings; and (5) fish eating frequency as independent variables. As a result, all the variables examined were significant and the explanatory powers (square of multiple correlation coefficient) of these variables were 17 or 7% for urinary Hg expressed as microgram Hg/l or microgram Hg/g Cr, respectively. Number of dental amalgam fillings or fish eating frequency explained only up to 1.5% of the total variance of urinary Hg. Contribution of Cr concentration, age and gender to urinary Hg level is also discussed. PMID- 8235574 TI - Ingested soil as a source of 137Cs to ruminants. AB - Ingestion of soil has been identified as a potentially important source of radionuclides, particularly to grazing ruminants. Soil artificially contaminated with radiocaesium was administered orally each day for 33 days to Bergamasca sheep and the transfer to milk measured. Two soil types were used, with a clay content of 11% and 16%, respectively. Transfer coefficient (fm) values to sheep milk, calculated at the equilibrium (period from 7th to 21st day after soil administration), were 3.72 x 10(-4) days l-1 (S.D. = 7.2 x 10(-5) days l-1) and 6.11 x 10(-4) days l-1 (S.D. = 9.1 x 10(-5) days l-1), respectively for the two soils. These low values indicate that radiocaesium is not removed from the binding sites on the clay minerals in the gut of the sheep. PMID- 8235575 TI - Frontiers in neuroscience. PMID- 8235576 TI - NCRR funding mechanisms. PMID- 8235577 TI - NIH support for graduate students. PMID- 8235578 TI - Environmental hazards: real or exaggerated? PMID- 8235579 TI - Environmental hazards: real or exaggerated? PMID- 8235580 TI - Carcinogenicity certification. PMID- 8235581 TI - New Sellafield study poses a puzzle. PMID- 8235582 TI - EMF-cancer links: yes, no, and maybe. PMID- 8235583 TI - Human embryo cloning reported. PMID- 8235584 TI - New protein appears to be long-sought neural inducer. PMID- 8235585 TI - Seeing the mind. PMID- 8235586 TI - Molecular genetics of neurological diseases. PMID- 8235587 TI - Excitement about calcium signaling in inexcitable cells. PMID- 8235588 TI - Thalamocortical oscillations in the sleeping and aroused brain. AB - Sleep is characterized by synchronized events in billions of synaptically coupled neurons in thalamocortical systems. The activation of a series of neuromodulatory transmitter systems during awakening blocks low-frequency oscillations, induces fast rhythms, and allows the brain to recover full responsiveness. Analysis of cortical and thalamic networks at many levels, from molecules to single neurons to large neuronal assemblies, with a variety of techniques, ranging from intracellular recordings in vivo and in vitro to computer simulations, is beginning to yield insights into the mechanisms of the generation, modulation, and function of brain oscillations. PMID- 8235589 TI - Neuronal mechanisms of object recognition. AB - Recognition of objects from their visual images is a key function of the primate brain. This recognition is not a template matching between the input image and stored images like the vision in lower animals but is a flexible process in which considerable change in images, resulting from different illumination, viewing angle, and articulation of the object, can be tolerated. Recent experimental findings about the representation of object images in the inferotemporal cortex, a brain structure that is thought to be essential for object vision, are summarized and discussed in relation to the computational frames proposed for object recognition. PMID- 8235590 TI - Programmed cell death and the control of cell survival: lessons from the nervous system. AB - During the development of the vertebrate nervous system, up to 50 percent or more of many types of neurons normally die soon after they form synaptic connections with their target cells. This massive cell death is thought to reflect the failure of these neurons to obtain adequate amounts of specific neurotrophic factors that are produced by the target cells and that are required for the neurons to survive. This neurotrophic strategy for the regulation of neuronal numbers may be only one example of a general mechanism that helps to regulate the numbers of many other vertebrate cell types, which also require signals from other cells to survive. These survival signals seem to act by suppressing an intrinsic cell suicide program, the protein components of which are apparently expressed constitutively in most cell types. PMID- 8235591 TI - Neural induction by the secreted polypeptide noggin. AB - The Spermann organizer induces neural tissue from dorsal ectoderm and dorsalizes lateral and ventral mesoderm in Xenopus. The secreted factor noggin, which is expressed in the organizer, can mimic the dorsalizing signal of the organizer. Data are presented showing that noggin directly induces neural tissue, that it induces neural tissue in the absence of dorsal mesoderm, and that it acts at the appropriate stage to be an endogenous neural inducing signal. Noggin induces cement glands and anterior brain markers, but not hindbrain or spinal cord markers. Thus, noggin has the expression pattern and activity expected of an endogenous neural inducer. PMID- 8235592 TI - Structure at 2.5 A of a designed peptide that maintains solubility of membrane proteins. AB - A 24-amino acid peptide designed to solubilize integral membrane proteins has been synthesized. The design was for an amphipathic alpha helix with a "flat" hydrophobic surface that would interact with a transmembrane protein as a detergent. When mixed with peptide, 85 percent of bacteriorhodopsin and 60 percent of rhodopsin remained in solution over a period of 2 days in their native forms. The crystal structure of peptide alone showed it to form an antiparallel four-helix bundle in which monomers interact, flat surface to flat surface, as predicted. PMID- 8235593 TI - Analysis of a nucleoprotein complex: the synaptosome of gamma delta resolvase. AB - The gamma delta resolvase protein is one of a large family of transposon-encoded site-specific recombinases. It performs recombination in a DNA-protein complex that contains 12 resolvase protomers and two copies of the 120-base pair DNA substrate, res (each with three binding sites for a resolvase dimer). A derivative of resolvase with altered DNA binding specificity was used to show that the role of resolvase at site I, which contains the crossover point, differs from its role at the other two binding sites. The resolvase dimers that initially bind to site I are the only ones that require the residue Ser10, essential for catalysis of DNA breakage. In addition, these site I-bound dimers do not use a specific interaction between dimers that is required elsewhere in the complex for synapsis of the res sites. PMID- 8235594 TI - Calcium sparks: elementary events underlying excitation-contraction coupling in heart muscle. AB - Spontaneous local increases in the concentration of intracellular calcium, called "calcium sparks," were detected in quiescent rat heart cells with a laser scanning confocal microscope and the fluorescent calcium indicator fluo-3. Estimates of calcium flux associated with the sparks suggest that calcium sparks result from spontaneous openings of single sarcoplasmic reticulum (SR) calcium release channels, a finding supported by ryanodine-dependent changes of spark kinetics. At resting intracellular calcium concentrations, these SR calcium release channels had a low rate of opening (approximately 0.0001 per second). An increase in the calcium content of the SR, however, was associated with a fourfold increase in opening rate and resulted in some sparks triggering propagating waves of increased intracellular calcium concentration. The calcium spark is the consequence of elementary events underlying excitation-contraction coupling and provides an explanation for both spontaneous and triggered changes in the intracellular calcium concentration in the mammalian heart. PMID- 8235595 TI - Microdomains with high Ca2+ close to IP3-sensitive channels that are sensed by neighboring mitochondria. AB - Microdomains of high intracellular calcium ion concentration, [Ca2+]i, have been hypothesized to occur in living cells exposed to stimuli that generate inositol 1,4,5-trisphosphate (IP3). Mitochondrially targeted recombinant aequorin was used to show that IP3-induced Ca2+ mobilization from intracellular stores caused increases of mitochondrial Ca2+ concentration, [Ca2+]m, the speed and amplitude of which are not accounted for by the relatively small increases in mean [Ca2+]i. A similar response was obtained by the addition of IP3 to permeabilized cells but not by perfusion of cells with Ca2+ at concentrations similar to those measured in intact cells. It is concluded that in vivo, domains of high [Ca2+]i are transiently generated close to IP3-gated channels and sensed by nearby mitochondria; this may provide an efficient mechanism for optimizing mitochondrial activity upon cell stimulation. PMID- 8235596 TI - Alterations of a zinc finger-encoding gene, BCL-6, in diffuse large-cell lymphoma. AB - The molecular pathogenesis of diffuse large-cell lymphoma (DLCL), the most frequent and clinically relevant type of lymphoma, is unknown. A gene was cloned from chromosomal translocations affecting band 3q27, which are common in DLCL. This gene, BCL-6, codes for a 79-kilodalton protein that is homologous with zinc finger-transcription factors. In 33 percent (13 of 39) of DLCL samples, but not in other types of lymphoid malignancies, the BCL-6 gene is truncated within its 5' noncoding sequences, suggesting that its expression is deregulated. Thus, BCL 6 may be a proto-oncogene specifically involved in the pathogenesis of DLCL. PMID- 8235597 TI - Isolation of the cyclosporin-sensitive T cell transcription factor NFATp. AB - Nuclear factor of activated T cells (NFAT) is a transcription factor that regulates expression of the cytokine interleukin-2 (IL-2) in activated T cells. The DNA-binding specificity of NFAT is conferred by NFATp, a phosphoprotein that is a target for the immunosuppressive compounds cyclosporin A and FK506. Here, the purification of NFATp from murine T cells and the isolation of a complementary DNA clone encoding NFATp are reported. A truncated form of NFATp, expressed as a recombinant protein in bacteria, binds specifically to the NFAT site of the murine IL-2 promoter and forms a transcriptionally active complex with recombinant protein fragment react with T cell NFATp. The molecular cloning of NFATp should allow detailed analysis of a T cell transcription factor that is central to initiation of the immune response. PMID- 8235598 TI - Spatial learning in mutant mice. PMID- 8235600 TI - Guam ALS-PDC: possible causes. PMID- 8235599 TI - Guam ALS-PDC: possible causes. PMID- 8235601 TI - Security of e-mail. AB - In Robert K. Englund's review of Before Writing by Denise Schmandt-Besserat (University of Texas Press, Austin, 1992) in the 11 June issue (p. 1670), two cuneiform characters in columns 1 and 2 on page 1671 were misrepresented. The correct characters are as follows. PMID- 8235602 TI - Hantavirus outbreak yields to PCR. PMID- 8235603 TI - The mouse-pinon nut connection. PMID- 8235604 TI - Women's health initiative draws flak. PMID- 8235605 TI - Cancer vaccines get a shot in the arm. PMID- 8235606 TI - In pursuit of protein folding. PMID- 8235607 TI - Hantavirus pulmonary syndrome: an emerging infectious disease. PMID- 8235608 TI - Multiple RNA polymerase conformations and GreA: control of the fidelity of transcription. AB - Pre-steady state kinetics of misincorporation were used to investigate the addition of single nucleotides to nascent RNA by Escherichia coli RNA polymerase during transcription elongation. The results were fit with a branched kinetic mechanism that permits conformational switching, at each template position, between an activated and an unactivated enzyme complex, both of which can bind nucleotide triphosphates (NTPs) from solution. The complex exists most often in the long-lived activated state, and only becomes unactivated when transcription is slowed. This model permits multiple levels of nucleotide discrimination in transcription, since the complex can be "kinetically trapped" in the unactivated state in the absence of the correct NTP or if the 3' terminal residue is incorrectly matched. The transcription cleavage factor GreA (or an activity enhanced by GreA) increased the fidelity of transcription by preferential cleavage of transcripts containing misincorporated residues in the unactivated state of the elongation complex. This cleavage mechanism by GreA may prevent the formation of "dead-end" transcription complexes in vivo. PMID- 8235609 TI - Guanidinium chloride induction of partial unfolding in amide proton exchange in RNase A. AB - Amide (NH) proton exchange rates were measured in 0.0 to 0.7 M guanidinium chloride (GdmCl) for 23 slowly exchanging peptide NH protons of ribonuclease A (RNase A) at pH* 5.5 (uncorrected pH measured in D2O), 34 degrees C. The purpose was to find out whether GdmCl induces exchange through binding to exchange intermediates that are partly or wholly unfolded. It was predicted that, when the logarithm of the exchange rate is plotted as a function of the molarity of GdmCl, the slope should be a measure of the amount of buried surface area exposed to GdmCl in the exchange intermediate. The results indicate that these concentrations of GdmCl do induce exchange by means of a partial unfolding mechanism for all 23 protons; this implies that exchange reactions can be used to study the unfolding and stability of local regions. Of the 23 protons, nine also show a second mechanism of exchange at lower concentrations of GdmCl, a mechanism that is nearly independent of GdmCl concentration and is termed "limited structural fluctuation." PMID- 8235610 TI - Formation of a molten globule intermediate early in the kinetic folding pathway of apomyoglobin. AB - Hydrogen exchange pulse labeling and stopped-flow circular dichroism were used to establish that the structure of the earliest detectable intermediate formed during refolding of apomyoglobin corresponds closely to that of a previously characterized equilibrium molten globule. This compact, cooperatively folded intermediate was formed in less than 5 milliseconds and contained stable, hydrogen-bonded secondary structure localized in the A, G, and H helices and part of the B helix. The remainder of the B helix folded on a much slower time scale, followed by the C and E helices and the CD loop. The data indicate that a molten globule intermediate was formed on the kinetic folding pathway. PMID- 8235611 TI - Detection of transient protein folding populations by mass spectrometry. AB - Hydrogen-deuterium exchange measurements are becoming increasingly important in studies of the dynamics of protein molecules and, particularly, of their folding behavior. Electrospray ionization mass spectrometry (ESI-MS) has been used to obtain the distribution of masses within a population of protein molecules that had undergone hydrogen exchange in solution. This information is complementary to that from nuclear magnetic resonance spectroscopy (NMR) experiments, which measure the average occupancy of individual sites over the distribution of protein molecules. In experiments with hen lysozyme, a combination of ESI-MS and NMR was used to distinguish between alternative mechanisms of hydrogen exchange, providing insight into the nature and populations of transient folding intermediates. These results have helped to detail the pathways available to a protein during refolding. PMID- 8235612 TI - Determination of type I receptor specificity by the type II receptors for TGF beta or activin. AB - Transforming growth factor-beta (TGF-beta) and activin signal primarily through interaction with type I and type II receptors, which are transmembrane serine threonine kinases. Tsk 7L is a type I receptor for TGF-beta and requires coexpression of the type II TGF-beta receptor for ligand binding. Tsk 7L also specifically bound activin, when coexpressed with the type IIA activin receptor. Tsk 7L could associate with either type II receptor and the ligand binding specificity of Tsk 7L was conferred by the type II receptor. Tsk 7L can therefore act as type I receptor for both activin and TGF-beta, and possibly other ligands. PMID- 8235613 TI - Interaction of Shc with the zeta chain of the T cell receptor upon T cell activation. AB - The shc oncogene product is tyrosine-phosphorylated by Src family kinases and after its phosphorylation interacts with the adapter protein Grb2 (growth factor receptor-bound protein 2). In turn, Grb2 interacts with the guanine nucleotide exchange factor for Ras, mSOS. Because several Src family kinases participate in T cell activation and Shc functions upstream of Ras, the role of Shc in T cell signaling was examined. Shc was phosphorylated on tyrosine after activation through the T cell receptor (TCR), and subsequently interacted with Grb2 and mSOS. The Src homology region 2 (SH2) domain of Shc directly interacted with the tyrosine-phosphorylated zeta chain of the TCR. Thus, Shc may couple TCR activation to the Ras signaling pathway. PMID- 8235614 TI - Gene replacement in Toxoplasma gondii with chloramphenicol acetyltransferase as selectable marker. AB - A system for stable transformation of Toxoplasma gondii tachyzoites was developed that exploited the susceptibility of Toxoplasma to chloramphenicol. Introduction of the chloramphenicol acetyltransferase (CAT) gene fused to Toxoplasma flanking sequences followed by chloramphenicol selection resulted in parasites stably expressing CAT. A construct incorporating the tandemly repeated gene, B1, targeted efficiently to its homologous chromosomal locus. Knockout of the single copy gene, ROP1, was also successful. Stable transformation should permit the identification and analysis of Toxoplasma genes important in the interaction of this opportunistic parasite with its host. PMID- 8235615 TI - Genetic identification of a hantavirus associated with an outbreak of acute respiratory illness. AB - A mysterious respiratory illness with high mortality was recently reported in the southwestern United States. Serologic studies implicated the hantaviruses, rodent borne RNA viruses usually associated elsewhere in the world with hemorrhagic fever with renal syndrome. A genetic detection assay amplified hantavirus specific DNA fragments from RNA extracted from the tissues of patients and deer mice (Peromyscus maniculatus) caught at or near patient residences. Nucleotide sequence analysis revealed the associated virus to be a new hantavirus and provided a direct genetic link between infection in patients and rodents. PMID- 8235616 TI - Alpha helix propensity of amino acids. PMID- 8235617 TI - Multifactorial nature of human immunodeficiency virus disease: implications for therapy. AB - The immunopathogenic mechanisms underlying human immunodeficiency virus (HIV) disease are extremely complex; the disease process is multifactorial with multiple overlapping phases. Viral burden is substantial and viral replication occurs throughout the entire course of HIV infection. Inappropriate immune activation and elevated secretion of certain cytokines compound the pathogenic process. Profound immunosuppression ultimately occurs together with a disruption of the microenvironment of the immune system, which is probably unable to regenerate spontaneously. Thus, therapeutic strategies in HIV disease must not be unidimensional, but rather must be linked to the complex pathogenic components of the disease and must address where feasible each of the recognized pathogenic processes for the possibility of therapeutic intervention. PMID- 8235618 TI - Dependence of calmodulin localization in the retina on the NINAC unconventional myosin. AB - Calmodulin is a highly conserved regulatory protein found in all eukaryotic organisms which mediates a variety of calcium ion-dependent signalling pathways. In the Drosophila retina, calmodulin was concentrated in the photoreceptor cell microvillar structure, the rhabdomere, and was found in lower amounts in the sub rhabdomeral cytoplasm. This calmodulin localization was dependent on the NINAC (neither inactivation nor afterpotential C) unconventional myosins. Mutant flies lacking the rhabdomere-specific p174 NINAC protein did not concentrate calmodulin in the rhabdomere, whereas flies lacking the sub-rhabdomeral p132 isoform had no detectable cytoplasmic calmodulin. Furthermore, a defect in vision resulted when calmodulin was not concentrated in the rhabdomeres, suggesting a role for calmodulin in the regulation of fly phototransduction. A general function of unconventional myosins may be to control the subcellular distribution of calmodulin. PMID- 8235619 TI - Crystal structure of neocarzinostatin, an antitumor protein-chromophore complex. AB - Structures of the protein-chromophore complex and the apoprotein form of neocarzinostatin were determined at 1.8 angstrom resolution. Neocarzinostatin is composed of a labile chromophore with DNA-cleaving activity and a stabilizing protein. The chromophore displays marked nonlinearity of the triple bonds and is bound noncovalently in a pocket formed by the two protein domains. The chromophore pi-face interacts with the phenyl ring edges of Phe52 and Phe78. The amino sugar and carbonate groups of the chromophore are solvent exposed, whereas the epoxide, acetylene groups, and carbon C-12, the site of nucleophilic thiol addition during chromophore activation, are unexposed. The position of the amino group of the chromophore carbohydrate relative to C-12 supports the idea that the amino group plays a role in thiol activation. PMID- 8235620 TI - An archimedian spiral: the basal disk of the Wolinella flagellar motor. AB - The motor that powers the rotation of the bacterial flagellum reaches through both membranes into the cytoplasm of Gram-negative bacteria. The flagellum is connected by a flexible link (hook) to the motor axis, which passes through the center of a structure called the basal disk. The basal disk functions with the L P ring complex as a bushing, enabling the rotation of the motor in the cell wall. The protein subunits of the basal disk of Wolinella succinogenes form an Archimedian spiral. The polymerization of subunits from a nucleation point at the motor in the form of a spiral allows constant growth of the basal disk. The disk is thought to provide a reinforcement at the flagellar insertion at the cell pole and to disperse forces that are generated by the momentum of the flagellar rotation. PMID- 8235621 TI - Replication-specific inactivation of the pT181 plasmid initiator protein. AB - Replication of the Staphylococcus aureus plasmid pT181, which occurs by the rolling circle mechanism, is accompanied by the covalent attachment of a approximately 12-residue oligodeoxy-nucleotide to one subunit of the dimeric plasmid-coded initiator protein, RepC. This oligonucleotide represents the plasmid sequence immediately 3' to the initiating nick site. The resulting heterodimeric protein lacks the topoisomerase and replication activities of unmodified RepC, suggesting that the regulation of plasmid DNA replication requires post-replicational inactivation of the initiator protein as well as control of its synthesis. PMID- 8235622 TI - Release of active cytokinin by a beta-glucosidase localized to the maize root meristem. AB - A beta-glucoside encoded by a cloned Zea mays complementary DNA (Zm-p60.1) cleaved the biologically inactive hormone conjugates cytokinin-O-glucosides and kinetin-N3-glucoside, releasing active cytokinin. Tobacco protoplasts that transiently expressed Zm-p60.1 could use the inactive cytokinin glucosides to initiate cell division. The ability of protoplasts to sustain growth in response to cytokinin glucosides persisted indefinitely after the likely disappearance of the expression vector. In the roots of maize seedlings, Zm-p60.1 was localized to the meristematic cells and may function in vivo to supply the developing maize embryo with active cytokinin. PMID- 8235623 TI - Rice prolamine protein body biogenesis: a BiP-mediated process. AB - Rice prolamines are sequestered within the endoplasmic reticulum (ER) lumen even though they lack a lumenal retention signal. Immunochemical and biochemical data show that BiP, a protein that binds lumenal polypeptides, is localized on the surface of the aggregated prolamine protein bodies (PBs). BiP also forms complexes with nascent chains of prolamines in polyribosomes and with free prolamines with distinct adenosine triphosphate sensitivities. Thus, BiP retains prolamines in the lumen by facilitating their folding and assembly into PBs. PMID- 8235624 TI - Formation and hydrolysis of cyclic ADP-ribose catalyzed by lymphocyte antigen CD38. AB - CD38 is a 42-kilodalton glycoprotein expressed extensively on B and T lymphocytes. CD38 exhibits a structural homology to Aplysia adenosine diphosphate (ADP)-ribosyl cyclase. This enzyme catalyzes the synthesis of cyclic ADP-ribose (cADPR), a metabolite of nicotinamide adenine dinucleotide (NAD+) with calcium mobilizing activity. A complementary DNA encoding the extracellular domain of murine CD38 was constructed and expressed, and the resultant recombinant soluble CD38 was purified to homogeneity. Soluble CD38 catalyzed the formation and hydrolysis of cADPR when added to NAD+. Purified cADPR augmented the proliferative response of activated murine B cells, potentially implicating the enzymatic activity of CD38 in lymphocyte function. PMID- 8235625 TI - Immune responses in interleukin-2-deficient mice. AB - The role of costimulatory signals in T cell induction was evaluated in mice lacking the interleukin-2 (IL-2) gene. In vitro secondary antiviral T cell responses were absent unless IL-2 was added, confirming the crucial role of IL-2 in vitro. In vivo, primary and secondary cytotoxic T cell responses against vaccinia and lymphocytic choriomeningitis virus were within normal ranges. B cell reactivity to vesicular stomatitis virus was not impaired. T helper cell responses were delayed but biologically functional. Natural killer cell activity was markedly reduced but inducible. These normal in vivo immune responses in IL-2 deficient mice question the importance of IL-2 as defined by in vitro studies. PMID- 8235626 TI - A Ca-dependent early step in the release of catecholamines from adrenal chromaffin cells. AB - Intense stimuli, such as trains of depolarizing pulses or the caffeine-induced release of calcium from intracellular stores, readily depress the secretory response in neuroendocrine cells. Secretory responses are restored by rest periods of minutes in duration. This recovery was accelerated when the concentration of cytosolic calcium was moderately increased and probably resulted from calcium-dependent replenishment of a pool of release-ready granules. Continuously increased concentrations of calcium led the over-filling of such a pool. Subsequently, secretory responses to stronger calcium stimuli were augmented. Hormone-induced calcium transients with a plateau phase of increased concentration of calcium may enhance the secretory response in this way. PMID- 8235628 TI - Minorities in science '93. Trying to change the face of science. PMID- 8235627 TI - Protection from HIV infection or AIDS? PMID- 8235629 TI - Minorities move from lab rats to policy wonks. PMID- 8235630 TI - What works. Building a global lab. PMID- 8235631 TI - Bringing science back to the neighborhood. PMID- 8235632 TI - Plenty of jobs, little minority support in biotech. PMID- 8235633 TI - AIDS activism and the OAR. PMID- 8235634 TI - Multiregional evolution. PMID- 8235635 TI - Jitters jeopardize AIDS vaccine trials. PMID- 8235636 TI - Popovic is cleared on all charges; Gallo case in doubt. PMID- 8235638 TI - Possible neandertal ancestor found. PMID- 8235637 TI - Medical ethics. A tough line on genetic screening. PMID- 8235639 TI - Tropical poison frogs. PMID- 8235640 TI - Misconduct in science. PMID- 8235641 TI - Misconduct in science. PMID- 8235642 TI - Can Russia slim down to survive? PMID- 8235643 TI - ORI drops Gallo case in legal dispute. PMID- 8235644 TI - Survey tracks misconduct, to an extent. PMID- 8235645 TI - German HIV-blood scandal reveals flaws in the system. PMID- 8235646 TI - Cell biologists explore 'tiny caves'. PMID- 8235647 TI - New Alzheimer's theory stirs controversy. PMID- 8235648 TI - The mice that roared. PMID- 8235649 TI - This is your brain on stress. PMID- 8235650 TI - Transgenic mice display a class (switching) act. PMID- 8235651 TI - The remaining stocks of smallpox virus should be destroyed. PMID- 8235652 TI - Why the smallpox virus stocks should not be destroyed. PMID- 8235653 TI - Checkpoints that couple gene expression to morphogenesis. PMID- 8235654 TI - Effective coupling in biological electron transfer: exponential or complex distance dependence? AB - Calculations for a simple model electron transfer system and tuna cytochrome c demonstrate a dichotomy in the distance dependence of the effective coupling. In one regime, the effective coupling varies exponentially with distance and depends primarily on the average properties of the bridging material; in the other regime, the effective coupling has a complex distance dependence and is more sensitive to the details of the bridging material. Experiments and theory indicate that both regimes may occur in biological systems, providing a perspective on a recent controversy over the nature of the distance dependence. PMID- 8235655 TI - Genetic relationships determined by a DNA heteroduplex mobility assay: analysis of HIV-1 env genes. AB - The genetic diversity of human immunodeficiency virus (HIV) is a major concern thought to impact on immunologic escape and eventual vaccine efficacy. Here, simple and rapid methods are described for the detection and estimation of genetic divergence between HIV strains on the basis of the observation that DNA heteroduplexes formed between related sequences have a reduced mobility in polyacrylamide gels proportional to their degree of divergence. Reliable phylogenetic subtypes were assigned for HIV-1 strains from around the world. Relationships between viruses were closest when derived from the same or epidemiologically linked individuals. When derived from epidemiologically unlinked individuals, the relationships between viruses in a given geographic region correlated with the length of time HIV-1 had been detected in the population and the number of strains initiating widespread infection. Heteroduplex mobility analysis thus provides a tool to expedite epidemiological investigations by assisting in the classification of HIV and is readily applicable to the screening and characterization of other infectious agents and cellular genes. PMID- 8235656 TI - Induction of metaphase arrest in cleaving Xenopus embryos by MAP kinase. AB - The natural arrest of vertebrate unfertilized eggs in second meiotic metaphase results from the activity of cytostatic factor (CSF). The product of the c mos(xe) proto-oncogene is thought to be a component of CSF and can induce metaphase arrest when injected into blastomeres of two-cell embryos. The c Mos(xe) protein can directly activate the mitogen-activated protein kinase kinase (MAP kinase kinase) in vitro, leading to activation of MAP kinase. MAP kinase and c-Mos(xe) are active in unfertilized eggs and are rapidly inactivated after fertilization. Microinjection of thiophosphorylated MAP kinase into one blastomere of a two-cell embryo induced metaphase arrest similar to that induced by c-Mos(xe). However, only arrest with c-Mos(xe) was associated with activation of endogenous MAP kinase. These results indicate that active MAP kinase is a component of CSF in Xenopus and suggest that the CSF activity of c-Mos(xe) is mediated by MAP kinase. PMID- 8235657 TI - Targeted insertion of a variable region gene into the immunoglobulin heavy chain locus. AB - A mutant mouse strain has been generated in which a rearranged immunoglobulin heavy (H) chain variable (V) region gene is placed into the heavy chain locus in its natural position, replacing the JH elements. In homozygous mutant mice, essentially all B cells in the spleen express the transgenic VH region in their antibodies. The proper location of the transgene relative to the constant region genes allows it to participate in isotype switching and undergo somatic hypermutation. Immunoglobulin transgenic mice generated in this fashion by gene targeting should prove useful for the exploration of immunoregulatory mechanisms. PMID- 8235658 TI - Generation of a mouse strain that produces immunoglobulin kappa chains with human constant regions. AB - Humanized antibodies are highly efficient as immunotherapeutic reagents and have many advantages over rodent antibodies. A mouse strain was generated by gene targeting to replace the mouse kappa light chain constant (C) region gene with the human C kappa gene. Mice homozygous for the replacement mutation (C kappa R) produced normal concentrations of serum antibodies, most of which carry chimeric kappa light chains, and mounted normal immune responses to hapten-protein conjugates. This technology provides a feasible option for the generation of high affinity humanized antibodies by means of the powerful somatic hypermutation selection mechanism. PMID- 8235659 TI - Bcl-2 inhibition of neural death: decreased generation of reactive oxygen species. AB - The proto-oncogene bcl-2 inhibits apoptotic and necrotic neural cell death. Expression of Bcl-2 in the GT1-7 neural cell line prevented death as a result of glutathione depletion. Intracellular reactive oxygen species and lipid peroxides rose rapidly in control cells depleted of glutathione, whereas cells expressing Bcl-2 displayed a blunted increase and complete survival. Modulation of the increase in reactive oxygen species influenced the degree of cell death. Yeast mutants null for superoxide dismutase were partially rescued by expression of Bcl 2. Thus, Bcl-2 prevents cell death by decreasing the net cellular generation of reactive oxygen species. PMID- 8235660 TI - Sensing structural intermediates in bacterial flagellar assembly by export of a negative regulator. AB - The ability of a regulatory protein to sense the integrity of the bacterial flagellar structure was investigated. In response to a defective hook-basal body complex, the anti-sigma 28 FlgM protein inhibits flagellin transcription. In cells with a functional hook-basal body complex, the flagellin genes are transcribed normally and the FlgM protein is expelled into the growth medium. In strains with a defective hook-basal body structure, FlgM is absent from the media. The presence of flagellin protein in the media is substantially reduced in strains carrying a FlgM-LacZ protein fusion, suggesting that the fusion is blocking the flagellar export apparatus. These results suggest that the FlgM protein assesses the integrity of the flagellar hook-basal body complex by itself being a substrate for export by the flagellar-specific export apparatus. PMID- 8235661 TI - False-positive human immunodeficiency virus testing in patients with lupus erythematosus. AB - Testing for human immunodeficiency virus (HIV) infection in patients with systemic lupus erythematosus (SLE) leads to a higher than expected rate of positive ELISA results and indeterminate Western blot results. Because most of these patients lack any risk factors for HIV infection and the coexistence of SLE and HIV infection is extremely rare, most of these results represent false positives. Two cases of false-positive HIV tests are reported and the related literature is reviewed. Further, Celum's algorithm for evaluating indeterminate HIV Western blot tests, which is especially valuable in lupus patients, is discussed. PMID- 8235662 TI - Course and prognostic value of Systemic Lupus Erythematosus Disease Activity Index in black Caribbean patients. AB - The course and prognostic value of disease activity measured by the validated Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) was investigated in 68 newly diagnosed black Caribbean cases. A high percentage of patients had clinical renal involvement (78%). Disease activity at onset was mild to moderate (SLEDAI < or = 10) in 36% of patients; about half never reached a higher SLEDAI score, whereas the other half advanced to higher disease activity (SLEDAI > 10). SLEDAI scores decreased significantly over time from diagnosis. Within 3 months after disease onset, 54% of patients reached their maximum SLEDAI scores. There were no differences in clinical features or survival between these patients and those with later (mean, 35 months) maximum disease activity, although the latter had more frequent disease flares. Overall survival was poor (91% and 56% at 1 and 5 years, respectively). High persistent disease activity (weighted average of SLEDAI scores > 10) was independently associated with decreased survival, whereas a high initial SLEDAI, a high maximum SLEDAI, and an increase number of flares were not. The main cause of death was infection, which often was associated with active disease (mean SLEDAI at death, 16 +/- 8.9). SLEDAI was a practical and reliable way of evaluating disease activity but was of limited prognostic value. PMID- 8235663 TI - Outcome measurement in scleroderma clinical trials. AB - Clinical trials in scleroderma were reviewed to assess the clinimetric properties of frequently used outcome measures. Twenty-seven controlled intervention studies were found in the English literature; nine demonstrated effective therapy. The outcome measures used included skin involvement, functional status, physical performance (grip strength, oral aperture), and internal organ involvement (pulmonary, gastrointestinal, renal, and cardiac). Very few outcome measures detected between- or within-group differences even when an active drug was compared with a placebo. Skin measures were found to yield statistical differences in seven studies, patient global assessment in three, and physician global assessments in four. Internal organ measures detected differences between groups only rarely; the pulmonary diffusing capacity was statistically different twice. Physical performance measures (eg, grip strength and oral aperture) never yielded statistical differences, and in only one of five trials did a functional assessment detect statistical differences. To show drug efficacy in future trials in scleroderma, better outcome measures need to be developed and a consensus obtained on which outcomes to use so that potentially effective therapies can be tested in a standardized fashion against a placebo or current therapy. Currently, because of a lack of clinimetric data on outcome measures, therapeutic inefficacy cannot be differentiated from a lack of sensitivity in the outcome measures used. In the future, outcome measures should be chosen on the basis of the adequacy of their reliability, construct, and content validity and be sensitive to change. Ideally, outcome measures also should have criterion validity, ie, show a strong association between the measure (such as a skin score) and an irrefutable gold standard (such as skin pathology). PMID- 8235664 TI - Comparative efficacy and safety of advanced drug therapy in children with juvenile rheumatoid arthritis. AB - Results from three randomized placebo-controlled trials were combined in a meta analysis to compare the clinical utility of four advanced drug therapy agents used to treat juvenile rheumatoid arthritis (JRA): D-penicillamine (10 mg/kg/d), hydroxychloroquine (6 mg/kg/d), auranofin (oral gold, 0.15 to 0.20 mg/kg/d), and two low dose levels of methotrexate [5MTX, 5 mg/M2/wk; 10MTX, 10 mg/M2/wk]. A total of 520 children with JRA were enrolled into these trials. Only 10MTX resulted in significantly greater improvement than placebo in variables that assess effectiveness: physician's global assessment, a composite index, and erythrocyte sedimentation rate. Treatment effect sizes were the largest in the 10MTX group for all articular disease indices. The short-term safety profiles were similar across all treatment groups. It is concluded that the current trend among pediatric rheumatologists to use oral methotrexate as the first advanced drug therapy in JRA is appropriate and that the minimum effective dose is 10 mg/M2/wk. PMID- 8235665 TI - Osteonecrosis: current perspectives on pathogenesis and treatment. AB - Nontraumatic osteonecrosis results from impairment of circulation to the affected bone. The femoral head is affected most frequently. The underlying cause for the circulatory defect in osteonecrosis varies and may involve both local and systemic changes. Steroid use, alcohol consumption, pancreatitis, and lipid disorders appear to lead to bone death either by development of fat emboli in the microcirculature surrounding the affected bone or by fatty infiltration of the marrow. Decompression syndrome results from the presence of gaseous emboli in the microcirculature. In Legg-Calve-Perthes disease other associated features are present such as short stature, suboptimal growth velocity, and/or hormonal imbalances, and it is likely that osteonecrosis may be secondary to systemic abnormalities, although specific factors have not been identified. Other frequently suggested pathogenic factors that play a role in the development of osteonecrosis include increased intraosseous pressures, the presence of cytotoxic cellular factors, intravascular coagulation, venous stasis, and the hyperviscosity syndrome. Some investigators have attempted, without success, to find a common etiology for all cases of osteonecrosis. In addition, patients have developed osteonecrosis without any known risk factors; this syndrome has been coined idiopathic avascular necrosis. In advanced stages of femoral head osteonecrosis, total hip arthroplasty appears to be the best therapeutic modality, particularly in older individuals. PMID- 8235666 TI - Bronchiolitis obliterans organizing pneumonia and rheumatoid arthritis. AB - The case of a 68-year-old woman who presented with dyspnea and upper lobe pulmonary infiltrates and shortly thereafter developed seropositive, erosive polyarticular rheumatoid arthritis (RA) is presented. An open-lung biopsy to evaluate progression of infiltrates showed bronchiolitis obliterans-organizing pneumonia (BOOP). Both lung and articular disease responded rapidly to corticosteroid therapy. Interrelationships between BOOP, bronchiolitis obliterans, and interstitial fibrosis with connective tissue disease are discussed. PMID- 8235667 TI - Pulmonary alveolar hemorrhage in systemic lupus erythematosus. AB - Pulmonary alveolar hemorrhage (PAH) is a rare and often fatal presenting feature of systemic lupus erythematosus (SLE). Reported mortality rates are extremely high, 70% to 90%. Death frequently occurs within the first several days of the hemorrhage. The hospital records of all inpatients with PAH and SLE between April 1986 and May 1991 at the Hospital of the University of Pennsylvania were reviewed. The complete resolution of PAH and the 75% survival rate found in this study is in marked contrast to previous reports. This experience suggests that patients with PAH and SLE may have a better prognosis than previously reported. PMID- 8235668 TI - [Suture protection of acute ruptured anterior cruciate ligament by the Pet-band (Trevira extra strong). Indications, technique results of a five-year study]. AB - In a prospective clinical study 56 acute tears of the anterior cruciate ligament (ACL) were treated between 1986 and 1991 by reinsertion and protection of the suture by means of a PET ligament (Trevira hochfest). The patients concerned were 31 men and 23 women (1 man and 1 woman had both knees operated on), with an average age of 39.7 years; 28 left and 28 right knees were affected. In 19 cases the ACL tear occurred in isolation, while in 37 concomitant intraarticular lesions were present. Nine patients had suffered multiple injury, and 3 had additional fractures distant from the knee. Haemarthrosis was encountered in 34 cases, and a clear effusion in 3 cases. The main causes of ACL rupture were sports injuries (n = 32), followed by traffic accidents (n = 14), activities of daily life (n = 6), and work accidents (n = 4). The preoperative diagnosis of ACL rupture was made correctly in 54 cases. After injury, 37 knees were operated on within the first 2 weeks, and 19 between the 3 and the 8 week. After arthroscopic repair of the concomitant lesions the alloplastic ligament was implanted isometrically by arthrotomy or miniarthrotomy through two bone tunnels and fixed to the bone with staples. Postoperative treatment included immediate continuous passive motion (CPM) and early weight-bearing with the protection of a brace. The patients were followed up at yearly intervals. At the last follow up, 6 years after the beginning of the study, 50 patients were examined clinically and radiologically, and the mean follow-up interval in these 50 was 40.2 months (12 79 months).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235669 TI - [Irrigation-suction drainage and synovectomy in the treatment of empyema of the knee joint]. AB - Following irrigation-suction drainage and synovectomy in 68 knee joints for empyema between 1971 and 1987, all were available for clinical and radiological follow-up. Since in 24 cases the empyema occurred after total knee arthroplasty, this group of patients was evaluated separately. The mean follow-up time in the no-prosthesis group was 8.3 and that in the prosthesis group, 5.3 years. In particular the aetiological factors, the bacterial pathogens and the effect of infection on radiological signs of degeneration were studied. The clinical results, including subjective findings, were assessed with the aid of HSS score. Our late results show that the prognosis for knee joint empyema can be rated as good if it is diagnosed early and treated aggressively. The poor outcome in a few cases was rather the result of underlying local disease and concomitant factors, and seemed not to be an immediate consequence of the infection itself. The relatively poor results in prosthesis group have to be referred predominantly to cases dating from the early years of total arthroplasty, with implants of the metal-on-metal type. PMID- 8235670 TI - [Minimal osteosynthesis of tibial pilon fracture]. AB - Fifty-eight patients with a total of 60 pillion fractures of the tibia were treated, mainly operatively, in the Department of Surgery (Klinikum Innenstadt, University of Munich) from 1984 to 1989. After 1-4 years, it was possible to question 90% of the patients, and 75% were also examined clinically and radiologically. The most frequent cause of fracture was a heavy impact, whereas sporting injuries accounted for only 13%. Thus, there was a high incidence (39%) of severe fractures (type C, AO classification) with tissue damage. The initial operative procedure was minimal osteosynthesis in 77% of the patients and tibial plate fixation in 20%. Delayed wound healing was observed in 17% with a rate of osteitis of 5%. The follow-up examination showed good to excellent results in 74%, and in two-thirds of the patients only minor arthrotic changes or none of all were found. Despite less severe fracture types in the group with tibial plate stabilization they more frequently had impaired wound healing (27% vs 14%) and severe posttraumatic arthrosis (43% vs 18%) than patients with minimal osteosynthesis. The results are discussed. PMID- 8235671 TI - [Long-term results of surgically treated Achilles tendon ruptures]. AB - The rupture of the Achilles tendon is a frequent sports injury. Between 1978 and 1988 we treated 358 men and 54 women, with an average age of 43 years, for this. In most cases the site of rupture was 3-5 cm proximal to the distal insertion of the tendon. The long-term subjective results of operative treatment of Achilles tendon ruptures (122 patients) were "good" or "very good" on follow-up examination in 85% of cases, and 97% of the patients treated surgically would choose the same therapy in the event of a new rupture. The isokinetic results demonstrated a (9.1%) loss of static and a 16.7% loss of dynamic strength in plantar flexion of the upper ankle joint compared with the healthy side. Ultrasound examination revealed thickening of the tendon itself and of the dorsal paratendon, with alterations in the tendon structure. In spite of the good results, the high rate of complications, 15.1%, shows that new and extensive studies on functional conservative therapy of Achilles tendon rupture are necessary to find whether conservative or operative therapy is better. PMID- 8235672 TI - [Simple elbow dislocation. Comparison of long-term results after immobilization and functional treatment]. AB - From 1976 to 1985 the treatment used for simple dislocation of the elbow consisted in closed reduction and immobilization for about 3-4 weeks. Since 1985, simple dislocation of the elbow has been treated with closed reduction and immobilization for fewer than 3 days. A group treated by immobilization (20 patients) and a group that underwent functional treatment (24 patients) were both evaluated with regard to elbow motion, instability, isometric flexor and extensor strength of the forearm, and subjective (patient) judgement of the elbow function. An average of 8 +/- 5 years after the treatment 39 elbows were re examined. In both groups, results were generally good, 22 patients having no residual symptoms or signs. However, 17 did have symptoms: 6 patients had a slight (loss of extension < 10 degrees, loss of flexion < 10 degrees) and 1 patient a moderate decrease in the range of motion (loss of extension > 10 degrees). Some instability was found in 5 patients and strength was slightly decreased in 9. The subjective judgement on their elbow function was very good in 22 patients. At follow-up, no statistically significant differences were found between the two groups. The retrospective study, however, revealed that immobilization for 3-4 weeks more than doubled the time of after-treatment, the period of disability of the elbow, and the period of physical rehabilitation. Therefore, an accelerated functional treatment is recommended for the treatment of simple elbow dislocation: this is more effective, takes less time and is more economical. PMID- 8235673 TI - [Sinus tarsi syndrome. Results of surgical treatment]. AB - Following exclusion of other causes of pain debridement of the sinus tarsi was carried out in 47 hindfeet with persisting pain despite local infiltration with steroids. In 41 patients with a total of 42 feet treated this way, evaluation by a research questionnaire was possible: 15 patients had some persistent problems or had inadequate results after surgery, and in 13 of these a clinical follow-up examination was carried out. Our retrospective study confirms that surgery is successful in treating chronic pain syndromes in the region of the sinus tarsi, as is known from the literature. It is especially successful if the pain is not due to an earlier fracture. In 60% of the hindfeet an instability required a ligamentous reconstruction is found on surgery. This high percentage suggests that the development of the sinus tarsi syndrome may quite often be due to an instability in the hindfoot. PMID- 8235674 TI - [Bilateral reversed palmaris longus muscle--a rare cause of peripheral median nerve compression syndrome. Case report]. AB - A rare case of median nerve compression syndrome outside the carpal tunnel in the distal forearm is reported. A 21-year-old man suffered while working from symptoms of temporary median nerve compression in both forearms; this was caused by hypertrophy of reversed palmaris longus muscles. Resection of the abnormal muscle bellies relieved the symptoms immediately. Only ten similar cases have been reported in the literature, and this is the first case with bilateral symptoms. PMID- 8235675 TI - [Local and systemic trauma in plate osteosynthesis of femur shaft fracture]. AB - Compression plating of fractures of the femur has a number of logistic advantages over intramedullary stabilization techniques though it is equally demanding for the surgeon. In this retrospective study of 77 cases, we try to show that compression plating of fractures of the femoral shaft is still a viable alternative to intramedullary fixation, especially in the light of local complications, postoperative systemic implications for the patient with multiple injuries, and the time required to achieve bone union. The data relating to all three of these aspects permit the conclusion that the results reached with compression plating of both simple and comminuted femoral shaft fractures are comparable to those reported following intramedullary nailing. Our treatment regimen does not involve a higher risk to the patient. PMID- 8235676 TI - [Tensile strength of various anchor systems in surgical correction of instability of the shoulder joint]. AB - Commercially available fixation devices used for reattachment of capsuloligamentous tissue to the glenoid rim were tested until they failed in 42 human cadaveric shoulders, and compared to the tension strength of the standard Bankart procedure and that of the intact anterior capsule. The bone quality in the glenoid was measured by quantitative computed tomography, which allowed selective quantification of cortical and cancellous quantification of cortical and cancellous bone. The mean load at failure perpendicular to the bone surface varied from 90 to 115 N and was lower than in the standard Bankart procedure (127 N). All suture anchors demonstrated similar holding strength with the exception of an absorbable wedge (P < 0.05). Due to reduced mineral content in the inferior part of the glenoid, the fixation strength of all suture anchors was significantly decreased. In more than 20% a tension load of 100 N caused the suture anchor to pull out, but in general the fixation strength was limited by the suture attachment and the quality of the soft tissues. These data together with the first clinical results of 49 cases of anterior instability operated on between 1988 and 1992 allow the conclusion to be made that this modification will provide sufficient stability for early mobilization after surgical repair. PMID- 8235677 TI - [Unilateral locking of cervical facet joints. Frequency and value of roentgenologic signs]. AB - Unilateral locking of cervical facet joints is often misdiagnosed and inadequately treated, because it is not readily detected on plain radiographs. Primary radiographs of 17 patients were analysed to evaluate radiographical signs with reference to frequency and significance. Direct signs of locking were present in no more than 53% of cases. Of these, an abrupt change in laminar space width, seen in 88.2%, proved to be the most reliable sign. Displacement of the spinous processes above and below the lesion was found in the same percentage of cases, but it carries less diagnostic weight. While present in all cases, anterolisthesis is only diagnostic if additional oblique views show evidence of lateralized cervical anterolisthesis. Clearly less reliable indirect signs included: the bow tie sign (29.4%), dehiscence of the spinous processes (23.5%), differences in the sagittal diameters of the vertebral bodies above and below the lesion (35.2%), double facet contour (47%) and tilting of the cranial segment of the cervical spine away from the locked facets (58.8%). Abnormalities of disc shape were not seen on a.-p. projections. Those detectable in axial views are irrelevant to locking, as are empty facets. If more than one indirect sign is present and if unilateral locking is suspected but cannot be established unequivocally even on additional oblique views, computed tomography is indicated, because reduction continues to be the first step in the management of fresh injuries. PMID- 8235678 TI - [Malignant non-Hodgkin's lymphoma of the joint. Report of two cases, review of the literature and problems of classification]. AB - This paper gives a survey of malignant Non-Hodgkin lymphomas (NHL) of joints. Primary and secondary malignant lymphomas of joints are rare. Our analysis of 6 years', new entries in the lymph node register in Kiel, covering 37.367 cases of malignant lymphomas, revealed only 20 cases (0.05%) with infiltration of joints. Slightly more men than women (1.18:1) are actually affected by articular lymphomas. In addition to 34 articular lymphomas of joints described in the literature between 1974 and 1992, we analyse 2 cases seen in our own institute and the 20 cases taken from the Kiel lymph node register. The peak incidence is between 40 and 60 years of age. One of our own cases, a woman of 92, is the oldest person ever affected. The most frequent localizations of NHLs are the knee, hip, and shoulder joints, with low-grade lymphomas predominating. Osteolytic lesions on radiographs are of decisive clinical and diagnostic significance for malignant articular lymphomas with affected bones. Lymphomas of joints are treated almost exclusively by radiation and chemotherapy, and sometimes a combination of both. Decisive prognostic significance attaches to the grade of malignancy and the fact of primary or secondary infiltration of the synovium and of the bone. The average survival time for patients with low-grade NHLs is 19 months with infiltration of the synovium and 27 months with infiltration of joint forming bones only, whereas the average survival time for patients high-grade lymphomas is 8 months with infiltration of the synovium and 11 months with infiltration only of joint-forming bones.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235679 TI - [Suture of the anterior cruciate ligament--what is the real value of this method?]. AB - After suture of the anterior cruciate ligament without anatomical augmentation, used as a routine method, 88 patients were followed up after a minimum of 2 years using the Lysholm score and the activity scale. Residual laxity was checked clinically and with the KT-1000. For comparison, 20 patients with chronic symptomatic anterolateral instability were followed up who had been operated on using the Eriksson technique during the same time period. Postoperative treatment was a limited functional approach. The peripheral ligament structures were treated following the Hughston and Muller principles. In two-thirds of the knees lateral tenodesis was added. The results of the suture method were disappointing and inferior to the results obtained by the Eriksson technique (trend). The Lysholm score was 77 and 82, respectively. The activity scale was 4.4 (recreational sports) for both groups. The Lachmann sign (KT-1000) in side-to side comparison (89 N) showed a residual laxity of 1.9 mm and 0.8 mm (average), respectively. In no series was tractopexy of demonstrable advantage. The final conclusion must be that the suture method (with iliotibial tract tenodesis) is not superior to guided conservative treatment. Despite an unfavorable situation preoperatively, the (abandoned) technique of ligament substitution is superior to the suture method (trend). In isokinetic testing dominant knees fare better. In good results, a high hamstrings-quadriceps ratio is typical. The reasons for this are not well-trained hamstrings, but the relative insufficiency of the quadriceps mechanism. PMID- 8235680 TI - [Primary suture of the anterior cruciate ligament. A critical analysis]. AB - Recently some authors have claimed that primary repair of an acute tear in the anterior cruciate ligament will fail in the long run. A review of the literature reveals that this opinion is mainly based on an American study in 1976. However, the poor results presented in this study were never reproduced. The study is critically analyzed, and 14 more papers presenting successful primary reconstructions of the anterior cruciate ligament are discussed. Preserving proprioceptive structures may be an important advantage of this technique, as it is hypothesized that the anterior cruciate ligament functions as a significant sensory organ, not only providing proprioceptive information, but also initiating protective and stabilizing muscular reflexes. In conclusion, it is recommended that primary suture be used combined with intra-articular semitendinosus tendon augmentation for rapid rehabilitation in cases of acute tears in the anterior cruciate ligament. PMID- 8235681 TI - [Nuclear magnetic tomography of the meniscus in asymptomatic probands]. AB - In 82 asymptomatic subjects aged 8-62 years we evaluated the menisci by MRI [1.0 tesla; SE (spin-echo) 700/20, PS (partial saturation) 500/10, STIR (Short-Time Inversion-Recovery-Sequence) 1600/130/30]. The meniscus was graded by means of a standard classification (grades 0-4). In SE sequences we found grade 0 in 45.5% of all cases, grade 1 in 33.2%, grade 2 in 16.1%, grade 3 in 4.5%, and grade 4 in 0.7%. In PS sequences we found grade 0 in 23.9%, grade 1 in 27.5%, grade 2 in 31.5%, grade 3 in 15.9%, and grade 4 in 1.1%. In STIR sequences we found grade 0 in 56%, grade 1 in 25.4%, grade 2 in 11.1%, grade 3 in 6%, and grade 4 in 1.6%. Correlation of meniscus degeneration with subjects' age showed relation of meniscus degeneration with subjects' age showed relation of meniscus degeneration with subjects' age showed an increase of grade 3 and grade 4 lesions with advancing age. Subjects older than 50 years presented grade 3 and 4 lesions in the SE sequence in 28.5% of the cases, in PS sequences in 40.7% of the cases, and in STIR sequences in 25% of the cases. MRI shows meniscal lesions in a significant proportion of asymptomatic patients, especially among subjects older than 50 years. The orthopedic surgeon has to test whether the clinical findings in patients with knee complaints coincide with the results of MRI. PMID- 8235682 TI - [Autologous transplantation of rib perichondrium in treatment of deep cartilage defects of the knee joint of sheep. Morphologic comparison of two resorbable fixation methods]. AB - The purpose of this study was to examine the fate of autologous perichondrial grafts after transplantation into cartilage lesions in weight-bearing joints, with reference to different fixation methods and different regimens of postoperative functional treatment. Full-thickness osteochondral lesions were drilled in the articular surface of knee joints in 36 sheep. The defects were filled with autologous rib perichondrial grafts, which were secured by either collagen sponges (group A; n = 12 animals) or fibrin glue (group B; n = 12 animals). Defects not filled with perichondrial grafts served as controls (group C; n = 12 animals). Following 1 week of immobilization of the treated leg, the plaster was removed and the animals were allowed to move freely. Animals were sacrificed after 4, 8, 12 and 16 weeks. The grafts were removed and investigated histologically. In contrast to weight-bearing areas and control defects, hyaline like cartilage formation could be noted in non-weight-bearing areas after 4 weeks. This newly formed cartilage revealed strong metachromasie following staining with acidic toluidine-blue and reacted positively with PAS, indicating de novo synthesis of proteoglycans and glycoproteins. Enzyme-histochemistry for alkaline and acid phosphatase activity showed positive reactivity only at the base of the transplants. Quantitatively, hyalin-like cartilage was achieved in 40/48 transplants from the non-weight-bearing area (20/24 transplants fixated with collagen sponge (group A); and 20/24 fixated with fibrin glue (group B). In control defects only fibrous scar tissue was detectable. In transplants from the weight-bearing area only 7/48 (24 of group A and B each) a differentiation into hyalin-like cartilage was visible.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235683 TI - [Late results of 351 femoral neck fractures (1986-1987 treatment period)]. AB - A total of 351 femoral neck fractures were treated from 1986 to 1987. The fractures were classified into three groups according to Pauwels. Nineteen percent of fractures were treated conservatively; in 65% internal fixation was performed, while in 16% total hip endoprosthesis was done. Of these patients 254 were reexamined three years later. In only 24.4% of the 254 patients were poor results--necrosis of the femoral head--found. PMID- 8235684 TI - [Radiologic determination of bone screw exit site on the femoral condyle. A method of exact imaging and length determination of internal fixation devices (screws, K-wires) in metaphyses of larger tubular bones exemplified by the distal femur]. AB - Distal femur imaging (A-P and lateral X-rays) after internal fixation often give only poor visualization of the exact position. A model with two anteriorly positioned cortex screws was used to show that an additional oblique X-ray, or fluoroscopy of the tip of the screw, allows much more exact determination of its length, so that it can be changed if necessary. This method is considered by the authors as a simple check of screw length measurement in "foyer fermee" procedures. PMID- 8235685 TI - [Ankle sprains. Comparison of long-term results of functional treatment methods with adhesive tape and bandage ("brace") and stability measurement]. AB - Both taping and bracing can be applied in the early functional treatment of ankle sprains. In this study the long-term (2.3 +/- 0.5 years) results of functional treatment with two types of bandages were compared. Out of 165 patients treated, 112 were available for interview, 60 of whom had been treated with adhesive, non elastic tape and 52, with a confection brace. In 93 of these patients (47 in the tape group), the ankle was examined and stabilometry was performed. The distance (D) and area (A) covered by each patient's centre of gravity while he/she stood on one leg for 30 s were measured. The following symptoms were found on the total population: pain on movement in 5%, swelling in 8% and functional instability (recurrent sprain or a feeling of giving way) in 38%. Mechanical instability was found in 34%. Pain on palpation of the lateral ligaments was the only symptom whose frequency differed significantly (P < 0.05) between the tape group (47%) and the brace group (20%). Stabilometric measurements revealed no substantial difference between the tape group (D: 436 +/- 100 mm; A: 192 +/- 87 mm2) and the brace group (D: 459 +/- 111 mm; A: 206 +/- 92 mm2). Nor was any difference in stability observed between the injured and the non-injured ankle, between the stable and the unstable ankle, or between the unstable ankle with and without brace application. Stabilometry is thus not an appropriate means of quantifying the symptoms of ankle instability.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235686 TI - [Impaired gait after base fractures of the 5th metatarsal bone]. AB - Between 1982 and 1990, twenty fractures of the base of the fifth metatarsal were treated operatively and followed up for a mean of 4.5 years (range: 1-9 years). All patients were evaluated clinically, roentgenographically and with kinetic gait analysis. A scoring system was used to record and evaluate the clinical and radiological data. Nine patients had an excellent result and nine a good result; one patient had a fair and one a poor result. Examination by kinetic gait analysis showed a gait asymmetry in eleven of the twenty patients, with decreased load on the formerly injured areas of the involved foot and increased load on the corresponding areas of the contralateral foot. However, only three patients had a clinically visible gait disorder. We believe that the gait disorder described is an automatically adopted movement pattern. Its cause lies in the initial pain after the trauma and during the postoperative care and sometimes continues even after the cessation of pain. Kinetic gait analysis allows quantification of asymmetry of gait and clinically non-visible load disorder. Therefore, pain and established gait asymmetry are clinically relevant in such patients because they can be treated specifically. PMID- 8235687 TI - [Vacuum sealing as treatment of soft tissue damage in open fractures]. AB - In 1992, 15 of 152 patients with open fractures were treated with vacuum sealing. Drainage tubes are inserted into polyvinyl foam, which is used to fill in the wound or tissue defect. Polyvinyl foam and adjacent skin are covered with a transparent polyurethane dressing which is impermeable to bacteria. The connection of the drainage tubes to a suction device, such as vacuum bottles, produces negative pressure in the polyvinyl foam, which means a high-contact zone of the foam-wound interface. This results in efficient cleaning and conditioning of the wound, with marked proliferation of granulation tissue. Bone infection did not occur in any of our 15 patients; 1 patient sustained a soft tissue infection due to an insufficient sealing technique. When the correct technique was applied the infection cleared up. PMID- 8235688 TI - [Documentation of injuries]. AB - The expert assessment of injuries due to criminal offenses is of great importance for any later legal proceedings. The initial medical examination of severe lesions is very often performed by clinicians; however, and when it which must be surgically shows that immediate surgical treatment is indicated later forensic interpretation is only possible if precise records documenting the nature of the wounds have been kept by the first doctor involved. The most frequent patterns of these injuries and relevant forensic aspects are discussed. PMID- 8235689 TI - [Therapy of severe liver trauma--is there an indication for liver transplantation?]. AB - The therapy of severe hepatic trauma requires complex surgical procedures. Our current surgical management of these injuries is described, including liver transplantation for the treatment of potentially lethal hepatic trauma. PMID- 8235690 TI - Fludarabine: pharmacokinetics, mechanisms of action, and rationales for combination therapies. AB - Knowledge of the pharmacokinetics of a drug is essential to the optimal design of the dose and schedule of chemotherapy protocols. As an extension, an understanding of the mechanism of drug action is necessary to construct the optimal strategy for combination chemotherapy. Nucleoside antimetabolites such as fludarabine are pro-drugs that must enter cells and be phosphorylated to the nucleoside triphosphate before they can elicit biologic activity. Thus, knowledge of the pharmacokinetics of the triphosphate in target cells and an understanding of the mechanisms by which this active form of the drug act are indispensable to the rational design of treatment protocols. This article reviews the essential elements of the pharmacokinetics and mechanisms of action of fludarabine to provide a rationale for combinations of fludarabine with other chemotherapeutic agents and anti-cancer modalities. PMID- 8235691 TI - Comparison of fludarabine, cyclophosphamide/doxorubicin/prednisone, and cyclophosphamide/doxorubicin/vincristine/prednisone in advanced forms of chronic lymphocytic leukemia: preliminary results of a controlled clinical trial. The French Cooperative Group on Chronic Lymphocytic Leukemia. AB - In a multicenter clinical trial conducted by the French Cooperative Group on Chronic Lymphocytic Leukemia (CLL) between June 1, 1990, and October 1, 1992, 183 patients with stage B CLL and 79 patients with stage C CLL were randomized to receive either cyclophosphamide/doxorubicin/prednisone (CAP) (71 stage B patients and 25 stage C patients), or cyclophosphamide/doxorubicin/vincristine/prednisone (CHOP) (56 stage B patients and 27 stage C patients), or fludarabine (FDB) (56 stage B patients and 27 stage C patients). The mean follow-up was 14 months (standard deviation, 7 months). At the 6-month follow-up examination, the results suggested that FDB was more effective than CAP and CHOP in patients with stage B disease (n = 183), with 19% of FDB-treated patients achieving "clinical and hematological remission" (CR) compared with 11% of the CHOP-treated patients and 7% of the CAP-treated patients (P = .08; 6 degrees of freedom; chi-squared test). The rates of partial remission (PR) and overall response (CR + PR) were, respectively, 75% and 94% for the FDB-treated patients, 64% and 75% for the CHOP treated patients, and 65% and 72% for the CAP-treated patients. However, in patients with stage C CLL (n = 77) at entry to the study, the remission status at 6 months showed slightly greater improvement in the CAP-treated group (n = 25), in which 84% of patients achieved remission (CR + PR) compared with 64% of patients in the FDB-treated group (n = 27) and 63% of patients in the CHOP treated group (n = 27). Further analysis of the study data may help to clarify the significance of these findings and to determine whether FDB improves survival in patients with advanced CLL. PMID- 8235692 TI - Fludarabine in low-grade lymphoma. AB - During the last 4 years, 88 patients with low-grade non-Hodgkin's lymphoma have received fludarabine, 25 mg/m2 daily for 5 days, repeated every 3 to 4 weeks. Fifty-one patients received fludarabine at recurrence or when the disease was deemed resistant to conventional treatment, 21 patients received the drug in the context of "minimal residual disease" in the hope of complete remission being achieved with a view to proceeding to myeloablative therapy (cyclophosphamide and total body irradiation) with autologous bone marrow transplantation, and 16 newly diagnosed patients received fludarabine as first-line therapy. Myelosuppression was the predominant toxicity, with 55% and 31% of previously treated and newly diagnosed patents, respectively, becoming neutropenic (neutrophils < or = 1.0 X 10(9)/L). The response rate (complete and partial response) was 44% for both patients with recurrent/resistant disease (20 of 45 evaluable patients) and for those with "minimal residual disease" (nine of 20 evaluable patients). In newly diagnosed patients, the response rate was 69% (11 of 16 patients). Five patients died of infection while neutropenic. These results confirm the activity of fludarabine in low-grade non-Hodgkin's lymphoma. Its precise role remains to be determined. PMID- 8235693 TI - Fludarabine single-agent therapy for relapsed low-grade non-Hodgkin's lymphomas: a phase II study of the German Low-Grade Non-Hodgkin's Lymphoma Study Group. AB - In a phase II study, 45 patients with advanced low-grade non-Hodgkin's lymphomas (NHLs) who had failed on or had relapsed after first-line chemotherapy were treated with a 5-day regimen of fludarabine, 25 mg/m2/d, by a 30-minute infusion. All patients were pretreated and had received one to 11 preceding regimens (median, three regimens). Histologic subtypes included 17 centrocytic/centroblastic NHLs, three centrocytic NHLs, 23 lymphoplasmocytoid immunocytomas, and one case each of peripheral T-cell and lymphocytic lymphoma. From 38 presently evaluable patients, 12 (31%) cases responded (five [13%] complete and seven [18%] partial remissions). Treatment-associated toxicity was mild to moderate, with myelosuppression comprising the major side effect. From the 12 complete and partial response patients, seven are currently in unmaintained remission for more than 12 months. These data indicate a high activity of fludarabine in heavily pretreated patients with low-grade NHL. Further investigations are warranted to assess the most appropriate usage for this highly promising agent at earlier stages of low-grade NHL therapy. PMID- 8235694 TI - Interferon alfa has become a standard in the treatment of chronic myelogenous leukemia. PMID- 8235695 TI - Interferon in the treatment of chronic myeloid leukemia. Proceedings of a workshop. February 4-7, 1993, Flims, Switzerland. PMID- 8235696 TI - Combination of chemotherapy and interferon alfa-2b in the treatment of chronic myelogenous leukemia. PMID- 8235697 TI - Treatment of chronic myelogenous leukemia with interferon alfa-2c: response rate and toxicity in a phase II multicenter study. The Austrian Biological Response Modifier (BRM) Study Group. PMID- 8235698 TI - United Kingdom Medical Research Council Randomized Trial of Interferon Alfa in Chronic-Phase Chronic Myelogenous Leukemia. PMID- 8235699 TI - Combination therapy for chronic myelogenous leukemia with interferon alfa and other cytokines. PMID- 8235700 TI - Interferon alfa and low-dose cytosine-arabinoside for the treatment of patients with chronic myelogenous leukemia in chronic phase. French CML Study Group. PMID- 8235701 TI - Evolving approaches with interferon alfa in chronic myelogenous leukemia. PMID- 8235702 TI - Regulation of the cytokine network by interferon: a potential mechanism of interferon in chronic myelogenous leukemia. PMID- 8235703 TI - Prospects of T-cell immunotherapy for cancer by peptide vaccination. PMID- 8235704 TI - Axl, a novel receptor tyrosine kinase isolated from chronic myelogenous leukemia. PMID- 8235705 TI - Adoptive immunotherapy in human and canine chimeras--the role of interferon alfa. EBMT Chronic Leukemia Working Party. PMID- 8235706 TI - Prophylactic use of interferon alfa after bone marrow transplantation for patients with chronic myelogenous leukemia at high risk of relapse: a pilot study. The Chronic Leukemia Working Party of the EBMT. PMID- 8235707 TI - Chronic myelogenous leukemia: progress in chemotherapy and evaluation of prognostic score 1. The German CML Study Group. PMID- 8235708 TI - Bone marrow transplantation from unrelated donors in chronic myelogenous leukemia. PMID- 8235709 TI - Autologous blood stem-cell transplantation and recombinant interferon alfa in chronic myeloid leukemia. PMID- 8235710 TI - Evaluating the effect of P210 BCR/ABL on growth of hematopoietic progenitor cells and its role in the pathogenesis of human chronic myelogenous leukemia. PMID- 8235711 TI - Recent progress in the elucidation of interferon alfa/beta and interferon gamma actions. PMID- 8235712 TI - The pathogenesis of cholestasis: past and future trends. Introduction. PMID- 8235713 TI - Bile acid metabolism and the pathophysiology of cholestasis. PMID- 8235714 TI - Hepatocellular transport of bile acids. PMID- 8235715 TI - Dubin-Johnson and Rotor syndromes: molecular basis and pathogenesis. AB - The Dubin-Johnson syndrome, as well as the related conjugated hyperbilirubinemia conditions found in sheep and in rats, are most likely caused by a congenital impairment of an ATP-dependent transport system specific for a variety of multivalent organic anions, including bilirubin diglucuronide. Bilirubin is thus conjugated but inefficiently secreted into bile. The symptoms are mild, possibly due to the presence of residual transport activity in the membrane catalyzed by separate transport proteins. Corollary effects include, at least in rats, a reduction of bile acid-independent bile flow due to the inhibition of biliary GSH transport by accumulating substrates of the defective transporter. Dark discoloration of the liver is evident in humans and sheep but not (or to a lesser extent) in the albino rats. The discoloration is due to deposition of a pigment formed from products of amino acid catabolism that cannot be secreted into bile. The relationship of further secondary effects, such as a change in the pattern of excreted coproporphyrins, to the primary defect is less well defined. The identification of the canalicular transporter or transporters for organic anions and a molecular characterization of the defects are in progress. PMID- 8235716 TI - Disappearing intrahepatic bile ducts: the syndromes and their mechanisms. PMID- 8235717 TI - Neonatal cholestasis: developmental aspects and current concepts. AB - Cholestatic disorders in infancy present the pediatric gastroenterologist, clinical investigator, and basic scientist with a unique model to probe the interfaces between the development and expression of essential functional elements in the liver and the extrauterine environment. This article has addressed some of the developmental features that define the process of bile formation in the neonate. Specific areas that limit effective maintenance of the extrahepatic circulation or interrupt the transition from an intrauterine to an extrauterine environment, including infectious diseases or metabolic disorders, have been identified. One may anticipate that a greater understanding of those features that are central to the processes of hepatic regeneration, the structural organization of the liver, and the mechanisms of hepatic injury will continue to make this an exciting area of investigation. PMID- 8235718 TI - Intrahepatic cholestasis of pregnancy: an estrogen-related disease. AB - In the recent years new avenues have been opened in the treatment of ICP, a complex disorder that seems to represent a maladaptation of some young and otherwise healthy women, to estrogens or other sex hormones. New drugs have been shown capable of providing promising therapeutic effects either on pruritus, the main distressing symptoms of cholestasis (such as epomediol, silymarin) or both on pruritus and some biochemical abnormalities (such as UDCA). Future clinical and experimental studies should provide better insight into the pathogenesis of cholestasis, the mechanisms of bile formation and secretion, and the metabolism of estrogens and other sex hormones and their alteration relationship to cholestasis, a disorder that is highly prevalent in humans. PMID- 8235719 TI - Pharmacologic treatment of cholestasis. PMID- 8235720 TI - [Effects of epidermal growth factor on growth and differentiation of rat granulosa cells]. AB - It was known that epidermal growth factor (EGF) plays an important role in the regulation of reproduction. The present study was undertaken to investigate the effect of EGF on the proliferation and differentiation of cultured rat granulosa cells. The results showed that EGF inhibited the 3H-TdR incorporation into DNA of granulosa cells, while the progesterone production was increased due to enhanced 3 beta-HSD activity. Radioreceptor assay (RRA) suggested that there were specific receptors for EGF on the granulosa cell with a Kd of 1.83 +/- 0.30 x 10(-8) mol/L and a Bmax of 1.75 +/- 0.29 x 10(4) sites/cell. Using method of immunohistochemistry, no EGF-like immunoreactivity was found in the granulosa cells at different age or different estrous cycle, but in the theca folliculi, interstitium and corpus luteum. These results suggest that EGF can regulate the growth and differentiation of the granulosa cells in the course of maturation of the folliculi and granulosa cells. PMID- 8235721 TI - [Pancreatic polypeptide in the control of beta-endorphin and prolactin release from rat anterior pituitary in vivo and in vitro]. AB - This study was designed to examine the possible effects of pancreatic polypeptide (PP) on beta-endorphin (beta-EP) and prolactin (PRL) release from rat anterior pituitary in vivo and in vitro. Injection of 0.5 microgram or 2.0 micrograms PP into the third ventricle of the brain (3rd v.i.) produced a significant decrease of the beta-EP and PRL resting secretion. 0.5 microgram PP (3rd v.i.) did not affect restraint stress-induced release of beta-EP, but partially lowered stress induced release of PRL. 2.0 micrograms PP (3rd v.i.) partially reduced restraint stress-induced release of beta-EP and completely suppressed stress-induced release of PRL. In order to investigate a possible direct action of PP on beta-EP and PRL secretion from the anterior pituitary gland, we incubated dispersed anterior pituitary cells with synthetic PP (0.05, 0.625 and 1.00 micrograms) for 1 n, the secretion of beta-EP was not affected at any dosage tested, but 0.625 and 1.00 micrograms PP significantly decreased the PRL secretion. These data indicate that PP may have an inhibitory role in the control of beta-EP secretion at the level of the hypothalamus, and an inhibitory role in the control of PRL secretion at the level of either hypothalamus or anterior pituitary. PMID- 8235722 TI - [Physiological properties of A delta and C fibres of superficial branch of radial nerve in human]. AB - By means of intraneural micro recording (INMR) and intraneural microstimulation (INMS), properties of receptive units innervated by A delta and C fibers of the superficial branch of the radial nerve were measured on humans. Thresholds of 24 A delta units to mechanical stimulation varied significantly; their receptive fields ranged from a single point to 75.4 mm2, with one to four receptive sensitive zones. A delta units by INMS evoked tingling with projected fields varying between 1.2 and 97 mm2. Receptive fields and projected fields of these units overlapped each other and showed a proximodistal size gradient (P < 0.05). The results suggest that A delta units have accurate localizing capacity. Eleven nociceptive units had higher threshold to mechanical stimulation. INMS of C nociceptive units evoked burning pain. Both the receptive fields and projected fields of most of these units showed overlapping indicating that C nociceptive units, to some extent, can localize noxious stimulation. PMID- 8235723 TI - [Studies on the descending projections from the pontine pneumotaxic region to the nucleus raphe magnus in cats]. AB - Experiments were done on 23 pentobarbital anesthetized adult cats. 67 units recorded from the parabrachial and Kolliker-Fuse nuclear complex (NPBM-KF) could be antidromically activated by electrical stimulation of nucleus raphe magnus (NRM). Of the 67 units, 7 were respiratory related units (6 inspiratory, 1 expiratory). These comprise 8% of the 87 respiratory units examined. The antidromic latencies ranged from 0.4 to 2.5 ms. After injection of WGA-HRP (wheat germ agglutinin-conjugated horseradish peroxidase) into the NRM, many retrogradely labelled neurons were observed in the NPBM-KF. These results showed that neurons in the NPBM-KF sent axonal projections to the NRM, which may be involved in the control of respiration as well as nociception. PMID- 8235724 TI - [Effects of angiotensin II, atrial natriuretic peptide and arginine vasopressin on activity of subfornical organ neurons in rat brain slices]. AB - The effects of angiotensin II (AG II), atrial natriuretic peptide (ANP) and arginine vasopressin (AVP) on 87 subfornical organ (SFO) neurons from 31 brain slices of rats were observed. After perfusing the brain slices with AG II (10(-7) mol/L, 3 min), spontaneous discharge rate of 40/55 (72.73%) neurons was significantly increased, while that of 3/55 (5.45%) was decreased and 12/55 (21.82%) neurons were non-responsive. The excitatory effects of AG II on neurons in SFO were completely blocked by AG II receptor blocker saralasin (10(-6) mol/L). As the brain slices were perfused with atrial peptide III (AP III) (10( 7) mol/L, 3 min), the firing rate of 7/17 (41.18%) neurons was significantly decreased, while that of 2/17 (11.76%) neurons was increased and 8/17 (47.06%) neurons were non-responsive. By perfusing brain slices with AVP (10(-7) mol/L, 3 min), the firing rate of 8/15 (53.33%) neurons was significantly increased, while that of 3/15 (20.00%) neurons was decreased and 4/15 (26.67%) neurons were non responsive. Twelve SFO neurons were successively perfused with three peptides. Among them, one was excited by both AG II and AVP, 3 were excited by AG II and inhibited by AP III, and the other one was excited by AVP and inhibited by AP III. The results suggest that the discharge rate of SFO neurons is affected by AG II, ANP and AVP. SFO may be one of the central regions in regulation of water balance and blood pressure. PMID- 8235725 TI - [Potentiation of morphine- and ohmefentanyl-induced analgesia by cholecystokinin receptor antagonists in rat]. AB - It has been reported that intrathecal (i.t.) injection of CCK-8 showed a marked antagonism to analgesic effects mediated by mu-opioid receptors in rat. The present study was performed to ascertain whether the blockade of endogenously released CCK-8 by potent and selective CCK-A antagonist devazepide and CCK-B antagonist L-365260 would affect opioid analgesia at the spinal cord level. A marked potentiation of the analgesic effect induced by morphine (4 mg/kg, sc) was produced by i.t. injection of 100 ng devazepide or 2.5 ng L-365260. Dose-response curves for the enhancement of the two drugs on morphine analgesia were bell shaped. Intrathecal injection of 66 ng devazepide or 1.25 ng L-365260 was also shown to potentiate the analgesic effect induced by the selective mu-opioid agonist ohmefentanyl (OMF) (32 ng, i.t.). The dose-response curves were also bell shaped. Devazepide or L-365260 per se produced no significant changes in rat tail flick latency (TFL). The above results are interpreted to mean that endogenously released CCK-8 in the spinal cord plays an antagonistic role to opioid analgesia, and it is the CCK-B receptors that mediate the anti-opioid effect since the dose of devazepide is 40-50 times higher than that of L-365260. PMID- 8235726 TI - [Further studies on the depressor effect of erythrocytic antihypertensive factor]. AB - We previously demonstrated the presence of a long-acting antihypertensive factor (AHF) in the erythrocytes of essential hypertensive subjects (EHS). The present investigation demonstrates further that the AHF is also capable of producing a rapid and transient blood pressure lowering effect in stroke prone spontaneously hypertensive rats (from 26.8 +/- 1.7 to 20.1 +/- 1.5 kPa by 10-30 s, P < 0.001). An even more stronger antihypertensive effect could be found if the AHF prepared from normal subjects or rats was used. We also found that some hypertensive factor was present in EHS plasma, but not in normal subjects. The results of the present paper suggest that AHF deficiency and higher level of pressor substance may play an important role in the development of essential hypertension. PMID- 8235727 TI - [Inhibitory effect of intraventricular administration of morphine on the hyperviscosity and elevation of blood pressure induced by stress in the rat]. AB - Experiments were performed on 99 Wistar rats. It was found that hyperviscosity and elevation of blood pressure (BP) could be induced by hanging and restraining conscious rats with their four limbs tied on a frame. These effects were unaffected by bilateral vagotomy. By intravenous injection of propranolol or phentolamine, elevation of BP could be reduced, while stress-induced hyperviscosity could only be reduced by propranolol (i.v.). Stress-induced hyperviscosity and elevation of BP could be inhibited by electroacupuncture applied to the right hind leg or microinjection of morphine into 4th ventricle of the brain. On the other hand, if opiate receptor antagonist naloxone was given into the 4th-ventricle, the stress-induced hyperviscosity and elevation of BP could no longer be inhibited by electroacupuncture. It is suggested that the hyperviscosity and elevation of BP induced by hanging and restraining are mediated by excitatory cardiovascular sympathetic outflow with the result of activation of adrenoreceptors. Activation of the opiate receptors in the hindbrain may be responsible for decrease in stress-induced hyperviscosity and elevation of BP and for the inhibitory effect of electroacupuncture of the right hind leg on stress-induced hyperviscosity and elevation of BP. PMID- 8235728 TI - [The stimulatory effect of dorsal vagal complex injection of pentagastrin on gastric motility in rats]. AB - Changes in antral motility induced by pentagastrin (G5) microinjected into the dorsal vagal complex (DVC) were investigated in fasting conscious rats. Antral motility was monitored by using chronically implanted force transducer sutured to the antrum. DVC injection of G5 markedly stimulated the amplitude of gastric contraction in a dose-dependent manner. The effect, being rapid in onset and long lasting, could not be mimicked by intravenous administration of G5 but could be markedly inhibited by microinjection of antigastrin serum into DVC. Atropine (100 micrograms.kg-1.h-1) and vagotomy could also abolish the G5 induced antral motility. No significant changes in plasma motilin were observed in portal venous blood after administration of G5 in DVC. These results suggested that DVC is an important site of action for G5 induced stimulation of gastric contraction as mediated by vagal cholinergic pathway. PMID- 8235729 TI - [Role of oxygen-derived free radicals in stress-induced gastric ulceration]. AB - In the present work, the role of oxygen-derived free radicals in the pathogenesis of the gastric mucosal injury induced by cold-restraint stress was studied in rats. The results were as follows. (1) In rats pretreated with superoxide dismutase (SOD), a scavenger of superoxide anions, or with dimethyl sulfoxide and mannitol, scavengers of hydroxyl radicals, the gastric lesions induced by stress became much less extensive. (2) The mucosal content of malondialdehyde, a metabolic product of lipid peroxides, was significantly increased during stress. (3) Histochemical study revealed that the gastric mucosa abounded in xanthine oxidase (XO), the enzymic activity of which was increased during stress. In the rats pretreated with allopurinol, to inhibit XO activity, the extent of gastric mucosal lesions was decreased significantly. These suggested that oxygen free radicals might be one of the important factors in inducing gastric mucosal injury during stress and the increase of XO activity might be responsible for the production of the radicals. PMID- 8235730 TI - [Effect of intracerebroventricular injection of caerulein on stress-induced gastric mucosal lesion in rats]. AB - The effect of caerulein injected into cerebroventricle on the stress-induced gastric mucosal lesion of rats with tied four limbs immersed in water was investigated. Caerulein (1.0 ng/rat) reduced significantly gastric mucosal lesion, decreased gastric acid content, but increased the gastric content of the mucus and PGE2. Under electromicroscope, morphological sign of hyposecretion in the parietal cells and hypersecretion in the mucus cells could be seen. The effect of caerulein could be prevented by intraventricular injection of naloxone or subcutaneous injection of indomethacin, but not by atropine, phentolamine and propranolol. The results indicate that the protective effect of caerulein on the gastric mucosa is mediated partly by central morphine receptors and partly by enhancement of endogenous PGE2. PMID- 8235731 TI - [The action of medullary tail-flick related neurons in electroacupuncture analgesia]. AB - Three types of cells, i.e., off-neurons, on-neurons and neutral neurons, could be recorded in the rostral ventromedial medulla (RVM) in light anesthetized rats. Just prior to the occurrence of tail flick elicited by noxious heat, the on-cells showed a burst of activity while the off-cells exhibited a cessation of discharge. No change of neutral cell activity related to tail flick was observed. Electroacupuncture at two "Ci Liao" points which resulted in inhibition of tail flick reflex affected the spontaneous activity of off- and on-cells in a different way, i.e, the discharge of most off-cells was significantly increased (P < 0.001), while that of on-cells showed no obvious change (P > 0.05). Tail flick related responses of the both cells were inhibited. These results provide the evidence that the off-cells may be the main efferent neurons in RVM involving in electroacupuncture analgesia. PMID- 8235732 TI - [Effect of stimulation of renal afferent nerves on plasma cortisol concentrations]. AB - The effect of stimulation of the renal afferent nerve (RAN) on plasma cortisol concentrations before and after lesions of paraventricular nuclei of hypothalamus (PVN) in chloralose-anesthetized cats was investigated. In animals with intact arterial baroreceptor stimulation of RAN did not significantly change the plasma cortisol concentration. However, the same stimulation elicited a significant increase in plasma cortisol concentration when the baroreceptor was denervated. Lesioning of bilateral PVN by kainic acid could abolish the cortisol response to stimulation of RAN. These data indicate that arterial baroreceptor reflex inhibited the cortisol response to stimulation of RAN and that PVN may be involved in the cortisol response to stimulation of RAN. PMID- 8235733 TI - The impact of a child death on marital adjustment. AB - One continuing concern in the sociological and psychological literature has been with the mental health consequences of stressful life events. Occasionally such stressful events have been linked to other outcomes such as a deterioration in the extent or quality of the relationship between a cohabiting (usually married) couple. This paper takes data from a longitudinal study of parents of an infant who has died (due to a Stillbirth, Neonatal Death or Sudden Infant Death), to determine whether the relationship between the parents is adversely affected. The follow-up data is available 2 months and 6-8 months after the infant death. The results indicate there is an increased marital break-up rate for parents whose infant has died. Further, shortly after the death there is evidence of a deterioration of the quality of the marital relationship between the partners whose relationship has remained intact. Both these consequences of an infant death are unlikely to be due to chance, but their magnitude is relatively modest. At the 6 month follow-up, there is evidence of a deterioration in the quality of the relationship for those partners whose infant survived, such that at 6 months there are no longer any marital adjustment differences between parents experiencing an infant death and parents whose infant survived. PMID- 8235734 TI - Social class gradients in years of potential life lost in Switzerland. AB - Analysis of the official Swiss mortality data have shown considerable social differences. In an earlier study of Swiss men aged 15-74 for the period 1979-1982 the social class mortality differentials have been analysed using standardized mortality ratios (SMR). The present study extends this previous work by calculating years of potential life lost (YPLL) before age 75, an additional indicator of mortality that puts more importance on deaths at younger ages. Emphasis is given on causes contributing to most years of life lost, especially to accidents and violent deaths, which result in more than 30% of total years of life lost. The distribution of years of life lost of the most important causes to social classes is illustrated also for age-specific groups. Additionally, this article presents all causes which account for more than 3% of total years of life lost. The social inequalities are shown as ratios between the social class with the highest (skilled manual workers) and the lowest risk (professionals). Most years of life are lost by skilled manual workers not only in general but also cause-specific. While the SMR from all causes of death showed a 2-fold difference between professionals and skilled manual workers, the social gradient in YPLL rate was even larger (2.5). Hence, the measure of years of potential life lost emphasizes the disadvantage of skilled manual workers to die earlier than professionals. The concept of YPLL proved to be a useful additional indicator not only of mortality in general, but also especially for monitoring causes, related to the lower social classes. PMID- 8235735 TI - Health behavior: issues, contradictions and dilemmas. AB - American medicine faces many contradictions and dilemmas. This is especially the case with regard to preventive health behavior. This paper explores the effects of several issues, contradictions and dilemmas on the American experience with primary preventive health behavior. These issues include: individualism, victim blaming, therapeutic nihilism, the over abundance of health information, America as a culture of risk takers, and the dilemma of the jungle vs the zoo. Four types of health behavior are defined. The first type of health behavior is the primary prevention of disease, defect, injury or disability. The second type is detection of asymptomatic disease, injury and defect. Third, is the promotion of enhanced levels of health, wellness and quality of life. And the fourth, at a more societal level, protective behaviors to make environmental transactions safe from disease, injury, defect and disability. These four types of health behavior are each explored in relation to societal values, technology and economics to determine which of these facilitate or impede health behavior at both the individual and societal levels. PMID- 8235736 TI - Variation in hemodialysis patient compliance according to demographic characteristics. AB - Patient noncompliance with treatment regime undermines the effectiveness of medical care, resulting in an unpredictable progression of the primary disease and a greater likelihood of complications. Hemodialysis patients are well suited for studying noncompliance because their treatment is prolonged and intensive, and medical regimens are clear cut and easily determined with objective measures. Moreover, noncompliant behavior by these patients not only endangers their life in the long run, but also results in negative effects within a day or two. Despite severe consequences, noncompliance with their medical regimen is the norm for dialysis patients rather than the exception. A large-scale study of chronic, outpatient hemodialysis patients (N = 1230 patients, response rate = 96%) treated in a variety of facility types (N = 29 facilities) was conducted to investigate prevalence and associated demographic characteristics of noncompliance with four treatment regimens--protein and potassium restrictions in the diet, medication regimen, and fluid restrictions. Few patients were noncompliant with diet regimens (9% with protein and 2% with potassium restrictions) but half were noncompliant with medication taking (50.2%) and fluid restrictions (49.5%). The prevalence of noncompliance with medication and fluid regimens was consistent with previous studies of dialysis patients but the prevalence of noncompliance with diet was much lower than that reported elsewhere. Prevalence of noncompliant patients varied between studies according to the cut-point used to establish compliance criteria, where more stringent cut-points inflated the percentage and more lenient cut-points reduced the percentage. Thus, there is a need to establish uniform criteria in order to test real differences in compliance between patient groups vs simply differences in measurement.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235737 TI - The hoarding habit, countertransference, and consultation anthropology in a Peruvian psychiatric hospital. AB - The 'hoarding habit' is the practice of collecting a large number of mostly useless objects by psychiatric patients. Countertransference consists of distorted perceptions by psychotherapists in their study of individuals, or social scientists in their study of human groups, which interfere with the pursuit of their therapeutic or research goals. A case is here presented from observations made by the author in one pavilion of the Hospital Victor Larco Herrera in Lima, Peru, of a 'bag man' who persisted in carrying his 'hoard' with him in large market bags. The practice of hoarding is related to the culture and social structure of the hospital, patients' use of physical space, the existence of trade networks, the smuggling of alcohol, and patients' needs to retain a feeling of selfness and personal autonomy as well as to maintain ties with the external world. In all these senses the hoarding habit is overdetermined: it is both a symptom of pathology and a sign of healthy functioning. However, its clinical construction may be expanded by an observer who can function as a 'culture broker' and who may be able to apply the dialectic of pathology and health to the operation of psychiatric services for more therapeutic ends. However, the observer, in a consultation capacity, is advised to go beyond the study of transference and countertransference in his or her subjects to focus on his/her own countertransference in order to consult more effectively. PMID- 8235738 TI - Inequalities in women's and men's ill-health: Britain and Finland compared. AB - This paper examines inequalities in ill-health among men and women in Britain and Finland, using national survey data from the mid-1980s. Age-standardised illness ratios are compared followed by multivariate logistic regression analyses. The degree of social inequality in ill-health for women and men is greater in Finland than in Britain. British employed women in each class report less limiting long standing illness than their Finnish counterparts. A major difference between the two countries is the poor health of British housewives. We relate these differences to societal variations in the participation of women in paid employment. In Finland women participate fully in paid work, whereas in Britain women are more likely to be full-time housewives or part-time employees. Unlike Finland, state provisions do not support the economic independence of British women. Structural variables, encapsulated by occupational class and employment status' are the primary factors associated with men's ill-health in Britain and Finland and also with Finnish women's ill-health. The difference between British and Finnish women is striking: class is associated with ill-health amongst women in both countries, but housing tenure and family roles are additional factors only among British women. In Britain, previously married women have particularly poor health. Our findings suggest that in a society such as Britain where paid employment is not universal for women, women's family roles and housing quality are associated with ill-health, but this is not the case in Finland, where women's participation in the labour market is near universal. PMID- 8235739 TI - Determinants of lactation performance across time in an urban population from Mexico. AB - We identified determinants of breastfeeding and full breastfeeding among 165 healthy mothers from Hermosillo, Mexico who planned to breastfeed and delivered vaginally a healthy term infant. Deliveries took place in either a nursery (n = 58) or a rooming-in (n = 107) public hospital where formula supplementation was not allowed. Multivariate analyses indicated that at 1 week full breastfeeding was associated (P < or = 0.05) with early milk arrival, social support for full breastfeeding and planned breastfeeding duration. At 2 months, full breastfeeding was associated with social support for full breastfeeding and early milk arrival. Breastfeeding was positively associated with early milk arrival and inversely associated with early introduction of supplementary bottles, maternal employment, maternal body mass index and infant age. At 4 months, full breastfeeding was positively associated with social support for full breastfeeding and inversely associated with infant age. Breastfeeding was positively associated with planned breastfeeding duration and inversely associated with early introduction of a bottle and urban background. Rooming-in mothers reported that their milk came in earlier (P < or = 0.05) than did the nursery group. Milk arrival was later when a bottle was introduced in the first week; both of these variables might be important in explaining a positive effect of rooming-in on lactation performance. PMID- 8235740 TI - Working with community organizations to evaluate potential disease clusters. AB - Increasingly, private citizens are organizing themselves to resist the intrusion of hazardous substances in their residential environments, or to promote the removal of the same. This movement is the source of the acronym (NIMBY-Not In My Back Yard). In North Carolina, the Cancer Surveillance Section has several experiences working with such community organizations. We have developed a commentary of our experience and observations on some of the characteristics that are more conductive to successful endeavors to resolve perception of increased health risk. Advantages and disadvantages of joint (health agency/concerned citizens group) studies or surveys are described. A case study of a successful joint survey to ascertain possible increased cancer incidence is described. PMID- 8235741 TI - Complexity, conflict, chaos, coherence, coercion and civility. PMID- 8235742 TI - On conflict resolution and change. PMID- 8235743 TI - A good word for the "young man (or woman) with a gun": whose violence? Whose "salutogenesis"? PMID- 8235744 TI - Sense of coherence and sociology of emotions. PMID- 8235745 TI - Alternative or additional medicine? An exploratory study in general practice. AB - In a South London general practice a substantial proportion of patients in some age groups (35 and 45 yr) had tried alternative treatment. Satisfaction with the outcome was limited. The use of alternatives was associated with higher general practice consultation rates, especially among women. Qualitative interviews with users suggested that rapid cures for chronic conditions were rarely expected; rather, an alternative explanation for health problems was sought and a greater sense of autonomy in dealing with them. Negative attitudes towards modern medicine arose from perceptions of excessive technological intervention and iatrogenic disorders arising from long term drug treatment. PMID- 8235746 TI - Dry and tight: sexual practices and potential AIDS risk in Zaire. AB - Both men and women in central Zaire like a 'dry, tight' vagina because it increases pleasure during sexual intercourse. In focus group interviews, they described wiping and washing procedures, as well as 30 different substances, mostly leaves and powders, that women can insert into the vagina to produce the desired effects. Women who use leaves said they crush them, insert them for several hours, then remove them before intercourse. Women who insert powders leave them in place during intercourse. Individual interviews with 99 women (half of them unmarried prostitutes and half married women) showed that over one-third of each group had used intravaginal drying or tightening substances at some time. Vaginal examinations by a physician revealed that several of the substances cause inflammatory lesions of the vagina and cervix. Furthermore, some products cause extreme dryness that could foster epithelial trauma during coitus, both for the woman and for her partner. Breaks in the epithelium may promote the passage of organisms that cause AIDS and other sexually-transmitted diseases. Thus the sexual practices of drying and tightening the vagina may be increasing the risk of infection. PMID- 8235747 TI - [Nutrition programs in a rural environment in the Philippines]. AB - The study of the relationship between the nutrition programs and the villagers of the Municipality of Plaridel, the target population, reveals that the Philippine Nutrition Program, in the process of planning and implementing the intervention programs, keeps apart from the community. As a result, the barangay people consider that these programs don't meet their needs and very few participate on them. Furthermore, in Plaridel, the intervention programs did not modify the nutritional status of the pre-school children, and the main short-term objectives of the Philippine Nutrition Program are far from being achieved. However, in this Municipality, there are social forces susceptible to become the supports of the intervention programs if these are based on the articulation of the recognized community needs and the articulation of the community's capacities. PMID- 8235748 TI - Economic and demographic research on malaria: a review of the evidence. AB - The primary objective of malaria control is to reduce deaths and illness from the disease. Elimination of infection is far more difficult. However, in most developing countries where malaria is endemic, there are few reliable data on levels, differences and trends in malaria mortality, and its relative importance as a cause of death or illness in endemic communities. The large-scale efforts and resources being devoted to delivery of essential care to mothers and young children in most countries make it important to have a better understanding of the extent of cause-specific child mortality levels in different epidemiological conditions so that appropriate interventions can be targeted at high risk groups and properly evaluated according to their contribution to mortality reduction. As per capita resources necessary in relation to the scale of the problem remain scarce, governments are finding that they must use their available resources more efficiently in meeting their health objectives. In the case of malaria, there are far too few empirical data documenting the marginal costs and benefits of alternative interventions and their response to an expansion (or contraction) of services, and to the level of endemicity and distribution of disease. Equally important, the literature on private costs of malaria (treatment, transportation etc) provides incomplete guidance to gains from control unless the authors begin to examine the policy implications of their results; for example, in many countries there is accumulating evidence that private expenditures outweigh public expenditures on health. What effect does this mix have on infectious diseases like malaria where public sector expenditures on prevention are so important in affecting incidence? PMID- 8235749 TI - Determining malaria effects in rural Colombia. AB - Good health is an integral component of the quality of human life, a prerequisite for developing human potential and an important determinant of economic development. When a person is ill from a tropical disease in an agricultural economy, a complex interaction between the individual's welfare and the family's welfare is set in motion. So complex are these interactions that few empirical studies exist on this subject and even where they do, empirical quantification of these interactions and economic losses places the analyst in the minefield of valuing time, ability and contribution to economic welfare. Placing monetary values on these commodities is always a little unsatisfactory since dollar values do not adequately reflect the nature of the losses. Secondly, the ill person's struggle to minimize the economic effects of disease on family income will mask its true impact; thirdly, tropical diseases disproportionately affect low-income groups and therefore measuring the income effects of disease amongst these groups will only reach at the earnings effect, and underestimate the economic implications of tropical disease control. Despite these difficulties, quantification of the economic impact of disease is important from a public health point of view. This study is an attempt at such a task, and focuses on the intra-familial struggle to minimize economic losses due to malaria. Using a case control approach, time-losses and labour reallocations within the household are examined in an attempt to understand the economic consequences of the disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235750 TI - The impact of malaria on labour use and efficiency in the Sudan. AB - The paper seeks to examine the economic impact of malaria in the Sudan. Using longitudinal empirical data from a farming population in the Gezira, it examines and quantifies labour losses through malaria within families, observing simultaneously the manner in which altruism or "nafeer" helps families and communities insure their members against disease and other consequences of uncertainty. PMID- 8235751 TI - Agricultural land use patterns and malaria conditions in Kenya. AB - This paper studies the problem of malaria control in irrigation and non irrigation areas in Kenya. Empirical results show that in both areas, households' level of awareness of malaria as a health problem, including its cause, was very high. However, attempts to trace the direct effects of malaria upon income or upon agricultural production were not statistically important. This does not imply that malaria has no consequence on household welfare. It is possible that the model equations were mis-specified--aggregate variables (total family size, total family income) and failure to quantify land in the production relationships may have contributed to these results. In addition, poor separation of malaria as a disease, from malaria as an infection, may have underestimated the effect of the disease on production. Thirdly, labour substitution (hiring or within-family substitution) was not measured in this early study, but was taken into account in subsequent research. Finally, labour requirements in the annual crop production schedules and the co-relation between these labour requirements and the pattern of adult morbidity were not longitudinally monitored. Cross-section data would bias the findings, particularly in those areas where the peak transmission season is short, where the crop grown does not require major labour input during this transmission season, and where acquisition of immunity would reduce the clinical impact of malaria upon adult labour. These vulnerabilities in the specification of the model and the data collected, probably affect the results obtained. Our empirical work raises a number of interesting and important questions which should be taken into account in future research. PMID- 8235752 TI - Economic and social consequences of malaria in new colonization projects in Brazil. AB - The success or failure of colonization in frontier areas of Brazil is an important question because of the country's declared commitment to undertake agrarian reform. However, demographic analyses of settler populations in new frontier areas suggests that these areas are not retaining agricultural settlers, but attracting transient wage-labourers in ranching, mining, construction and entrepreneurial activities such as placer mining or small businesses. One reason for this trend is the high prevalence of endemic diseases such as malaria within frontier areas. High malaria prevalence interferes with the scope and stability of permanent, agricultural settlement and this imposes economic and social costs which extend far beyond immediate need for control and treatment of the disease. PMID- 8235753 TI - Malaria rates and fate: a socioeconomic study of malaria in Brazil. AB - This paper examines behavioural risk factors for malaria in the Machadinho resettlement area in the Amazonian forests of Brazil. Analysis suggests that economic status and knowledge of the importance and behaviour of the mosquito in transmitting malaria are significant factors in determining prevalence risk, irrespective of whether preventive precautions (DDT spraying of houses, and clearing vector breeding sites) are undertaken in the endemic area. However, a higher economic status combined with better knowledge of the vector and DDT spraying decreases the risks of infection considerably. The results suggest that economic status--which is not easily subject to intervention--plays a more important role in transmission than is normally suspected, although preventive actions diminish the disease burden significantly. One might conclude that the landless and impoverished migrants who seek income, and independence in the jungle are destined to have malaria as one of their many burdens. A more positive implication is that control programmes must work harder and more intensively on behalf of poorer migrants in order to diminish the disease burden for these groups. PMID- 8235754 TI - Malaria and mobility in Thailand. AB - This paper examines the relationship between malaria transmission and migration in three northern Thai villages. Data and observations indicate that land-poor families forced into swidden farming have greater contact with the primary vectors in Thailand--which breed in small pools in forested areas and shady clearings on hilly scrub terrain. Once infected, migrants from an endemic locus can introduce the parasite into an area with no transmission but potent vectors, thus becoming the cause of explosive epidemics; equally, non-immunes carrying out agricultural activities in or across forest and border areas can themselves be subject to seasonal morbidity. In addition to agricultural activities on clearings near forested areas, clandestine forest activities and cross border traffic contributes to the high prevalence of malaria in Thai border villages. Illegal economic activities--logging, poaching, cattle and goods smuggling- interferes with vector suppression campaigns and prompt detection of infected cases, and ultimately increases human infection not only within the mobile population, but also within the passive population of villages to which the migrants return periodically. Control measures therefore need to take into account the economic pressures which determine a high degree of mobility, the ethnic diversity of the groups which depend on fringe activities for their economic welfare, and the difficult geography of the areas in which they live. As long as economic circumstances forcing human-vector contact receives inadequate attention, better alternatives to current vector control campaigns (which are not effective amongst migrants) are not tried and malaria transmission continues. PMID- 8235755 TI - Human mobility and malaria risk in the Naya river basin of Colombia. AB - This paper develops and tests a methodology for examining the manner and the extent to which human mobility can be associated with malaria risk within an endemic area. The paper presents entomological, epidemiological and mobility results obtained from macro data and micro data. Results indicate that malaria is concentrated in the delta zone of the Naya river basin where the circulation of people into this zone for economic reasons occurs at a time when vector densities are high. High human density combined with high vector density ensures continuous and intense transmission of malaria. In attempting to measure intensity of transmission, microscopic analysis of parasitaemia was found to underestimate prevalence showing it to be very low or negligible, whereas serological data collected during the same period of time reveal that prevalence is high, and similar for both sexes. Serological data also indicate that prevalence increases with age particularly from adolescence onwards. Since mobility for agricultural purposes involves able-bodied males and females, there seems ample justification for concluding that human mobility into certain areas increases the risk of infection for adults, particularly when such mobility coincides with peak rainfall and vector densities. Micro-level data are critical in pinpointing causation and small area variations in risk. Thus the beach area is a high risk area within the delta region, particularly for adults, and this is likely to be because of high vector biting rates in the beach environment. One conclusion is that micro analysis of behaviour, when linked to epidemiological data are particularly useful in defining and locating areas and groups at highest risk; using such information to achieve intensive coverage on a small scale may be more cost-effective than attempting to achieve vector or chemotherapeutic control over a wider area for all groups. PMID- 8235756 TI - Illness perception: social cost and coping-strategies of malaria cases. AB - This paper looks at illness perceptions and lay management of malaria in the context of new settlement. While the new settlement places unique pressures on the population there are several trends observed that have relevance for program planning and implementation. PMID- 8235757 TI - [From symptomatology to the person: the role of nursing care]. PMID- 8235758 TI - [When the patient becomes the symptom]. PMID- 8235759 TI - [The accident as symptom]. PMID- 8235760 TI - [Obstacles in the path of recollection: symptomatology and transference]. PMID- 8235762 TI - [Current issues in corporal practices]. PMID- 8235761 TI - [A music therapy workshop for demented persons]. PMID- 8235763 TI - [The group phenomenon in a society]. PMID- 8235764 TI - [Symptomatology in psychiatry: importance and usefulness]. PMID- 8235765 TI - [Daily care of profoundly retarded adults]. PMID- 8235766 TI - [Medicalization, symptomatology or defense mechanism]. PMID- 8235767 TI - Human herpesvirus 6. AB - Human herpesvirus 6 (HHV-6) is a relatively recently discovered virus. Although systematic studies on the spread of this virus are lacking, considerable data have been collected regarding the prevalence of HHV-6 infection. A number of laboratory methods are available for the diagnosis of HHV-6 infection, but currently these tests are usually available only in research laboratories. HHV-6 has been implicated in a number of infectious and lymphoproliferative and malignant diseases. While the relationship between HHV-6 and human immunodeficiency virus is not yet clear, HHV-6 has been shown to cause exanthema subitum and acute febrile illness in young children. At this time no adequate antiviral therapy is available for HHV-6 infection. PMID- 8235768 TI - Analgesic rebound headache: experience in a community hospital. AB - In the past, chronic daily headaches have been attributed to many possible causes, including "tension," excessive contraction of pericranial muscles, and depression. A careful history reveals that many patients start out with fairly typical intermittent migraine attacks that evolve over the years into a pattern of daily or almost daily head pain. Overuse of analgesics is usually a part of this clinical picture, which has been termed "analgesic rebound headache." Fifty three of the first 100 patients admitted to a community hospital's inpatient headache treatment program had chronic daily headaches and were overusing analgesics or other headache relief medications. We report our retrospective review of these patients and describe treatment of this difficult and sometimes unrecognized syndrome, primarily with intravenous dihydroergotamine (DHE). Results at hospital discharge and at follow-up several months later showed significant reduction of pain. PMID- 8235769 TI - Compensation status as a predictor of outcome in nonsurgically treated low back injury. AB - Whether the compensation status of patients injured in occupational accidents influences treatment outcome remains a controversial issue. This article describes the treatment outcome status of compensated versus noncompensated patients who received comprehensive functional restoration treatment of low back pain in a hospital-based, interdisciplinary, occupational rehabilitation and pain management program. Results of this investigation revealed treatment outcome differences between the two groups in two of three measures at discharge (subjective pain intensity and return-to-work), and outcome differences in one of five measures at 6-month follow-up (subjective pain intensity). Interestingly, significant group differences in return-to-work rates noted at the time of treatment discharge were not found during the follow-up period, with no group outcome difference in return-to-work rates noted at 6-month follow-up. PMID- 8235770 TI - Interhospital transfers, the COBRA Law (1985), and the use of a public assistance health plan. AB - Public outcry regarding unsafe interhospital patient transfers led to the passage of the COBRA Law of 1985. Since its implementation in 1986, the law has been sharply criticized for its sporadic enforcement, narrow focus, and lack of care provisions for the poor. We reviewed retrospectively the charts of all emergency department patients transferred from two Milwaukee private hospitals during two identical 6-month periods in 1985-1986 and 1988-1989. We compared demographic and clinical characteristics of the patients transferred during the two periods. The emergency departments cared for 15% of Milwaukee's emergency patient population and transferred 216 and 200 patients during each respective period. Uninsured transfers fell from 32% to 17% of the study populations. Although the proportion of the local population belonging to a health maintenance organization (HMO) did not change, transfer of HMO patients increased from 14% to 27%. Transfers to the area's various tertiary care centers made up 21% and 35% of all transfers and were conducted without regard to insurance status. There was a fourfold increase in the use of the county's public assistance plan for emergency hospitalization of indigent patients at local hospitals. Although the number of uninsured patients transferred from the study hospitals decreased markedly, this decrease was counterbalanced by increased private hospital use of the county's emergency hospitalization plan for indigent patients. This successful plan bears further examination by health policy planners as an important model for the provision of emergency hospitalization for the indigent. PMID- 8235771 TI - Osteomyelitis of the jaw in patients infected with the human immunodeficiency virus. AB - Eight patients with osteomyelitis of the jaw were identified at two county hospitals over a 4-year period. Three patients (37.5%) were found to be HIV seropositive, including two patients not previously known to be HIV-seropositive. Signs and symptoms of infection were similar in both groups of patients, and commonly included fever, pain, and swelling. Radiographs showed specific findings of osteomyelitis in one of three HIV-seropositive patients and four of four seronegative patients with chronic jaw infection. HIV-seropositive patients appeared to have a worse clinical outcome than their seronegative counterparts. We conclude that osteomyelitis of the jaw may be the presenting manifestation of HIV infection, and that careful attention and close follow-up should be applied to such patients because of their poor overall clinical response. PMID- 8235772 TI - End-stage renal disease with venous occlusion in both upper extremities. AB - We studied 27 patients with venographically documented bilateral subclavian vein occlusion thrombosis after the use of dual lumen hemodialysis catheters. Immediate venous access was severely limited because all patients had previously had multiple access failures. The rate of venous pressure rise between hemodialysis treatments is inconsistent in suggesting imminent thrombosis as the sole explanation for venous thrombosis, yet it is a signal to investigate vein patency. Venography may be the best method, pending further development of duplex ultrasonography. The lack of clinical symptoms warrants early use of duplex ultrasonography and venography to document subclavian vein occlusion thrombosis so that heparin and thrombolytic therapy may be initiated. Length of time between onset of pressure increase and initiation of therapy did not affect successful outcome. PMID- 8235773 TI - Pyogenic hepatic abscess. AB - We reviewed the clinical courses of 37 patients treated for pyogenic liver abscesses. These abscesses were cryptogenic in 38% of the patients, associated with biliary tract or intra-abdominal disease processes in 48%, and hematogenous in 11%. Computed tomography was more accurate (96%) in diagnosing a hepatic abscess than ultrasonography (82%). The majority of the patients were treated initially with percutaneous drainage (18) or needle aspiration drainage (5); treatment failure occurred in 9 of these 23 patients (39%). In contrast, there were no treatment failures in the 9 patients treated with primary operation. Overall, 4 of the 37 patients died (a mortality rate of 11%) but 2 of them died of underlying malignant disease rather than the liver abscess. PMID- 8235774 TI - Eustachian tube dysfunction and its sequelae in patients with cleft palate. AB - The incidence of middle ear disorders in patients with cleft palate deformities was determined in a retrospective review of 110 children with palate or palate and lip involvement. Ages ranged from 2 months to 18 years and included patients with associated abnormalities such as Pierre Robin syndrome and Treacher Collins syndrome. Clinical manifestations of eustachian tube dysfunction were found in 79% in the form of effusions, retraction pockets, adhesions, and ossicular erosion. Only two patients had confirmed cholesteatoma formation. Despite the low incidence of acquired cholesteatoma in these patients, the frequent occurrence of other disorders that affect hearing warrants early and aggressive otologic management to prevent long-term sequelae. PMID- 8235775 TI - Evaluation of flumazenil for reversing the effects of midazolam-induced conscious sedation or general anesthesia. AB - To evaluate the effectiveness of flumazenil in reversing midazolam-induced conscious sedation and general anesthesia, we gave either flumazenil or placebo to 55 patients in a double-blind manner after surgery. Whether surgery was done under conscious sedation (CS group) or general anesthesia (GA group) depended on the procedure. Recovery was assessed by an Observer Assessment of Alertness and Sedation (OAAS) Scale, Finger-Nose (F-N) test, and picture recall and recognition. OAAS and F-N tests were done at baseline, 0 minutes (before the test drug administration), and at 5, 15, 30, 60, 120, and 180 minutes. Picture recall and recognition were tested at 180 minutes and 24 hours, respectively, after test drug administration. Patients who were sedated during surgery (CS group) received a mean of 10.3 +/- 5.3 mg midazolam. The patients receiving general anesthesia (GA group) were given 21.0 +/- 8.2 mg midazolam. In the CS group, improvement in scores on the OAAS scale and F-N test was similar after administration of both flumazenil and placebo. In the GA group, flumazenil produced significantly greater improvement in the OAAS scores at 5 and 15 minutes, and in the F-N test scores at 15 minutes. In both groups, picture recall and recognition improved significantly immediately after flumazenil administration, but this improvement was generally not sustained for pictures shown at later times. These results imply that flumazenil is beneficial for reversing amnesia briefly after midazolam induced sedation. However, flumazenil hastens recovery only when larger doses of midazolam are used for general anesthesia. PMID- 8235776 TI - Pseudoseizures: etiologic and psychotherapeutic considerations. AB - Pseudoseizures are a somatic form of communication involving dissociative mechanisms. This paper outlines basic strategies for psychotherapeutic management with a case example. PMID- 8235777 TI - Efficacy of radiographic evaluation of the cervical spine in emergency situations. AB - Evaluation of trauma to the cervical spine usually includes predominantly normal x-ray studies that prevent expeditious workup and treatment and increase cost. In a retrospective review of the cases of 1,686 consecutive trauma patients who had routine radiographs for possible injury to the cervical spine, injuries were detected in 32 (1.9%). In a similar consecutive group of 597 patients whose cases were more critically analyzed prospectively, 17 (2.8%) had suspected injuries; of these, five (0.8%) were true positives. All patients in the prospective group with suspected injuries had neck pain, neurologic deficit, or altered consciousness, suggesting that emergency radiographic screening according to more selective clinical criteria may be feasible. PMID- 8235778 TI - Staging ovarian cancer diagnosed during laparoscopy: accuracy rather than immediacy. AB - Immediate staging and debulking of an unsuspected ovarian malignancy detected at the time of diagnostic laparoscopy is appropriate when personnel knowledgeable in these procedures are available. However, when assistance is unavailable, termination of the diagnostic laparoscopy and timely referral is acceptable. This report reviews techniques to preoperatively distinguish a benign from a malignant adnexal mass, steps to evaluate an adnexal mass during laparoscopy, and ovarian cancer staging procedures. PMID- 8235779 TI - Elective combined abdominal-vaginal cesarean hysterectomy for cervical neoplasia. AB - From January 31, 1990 to November 15, 1991, I treated three patients by cesarean hysterectomy with cervical conization for cervical neoplasia. All three hysterectomies were facilitated by the use of a combined abdominal-vaginal technique, which is described in this report. PMID- 8235780 TI - Incidence and simple management of priapism following dynamic infusion cavernosometry-cavernosography. AB - Of 239 patients with erectile dysfunction (aged 36 to 70 years) who were evaluated with dynamic infusion cavernosometry-cavernosography, 32 (13.4%) developed priapism after the procedure and were successfully managed with immediate intracorporal injection of phenylephrine. No single risk factor for the development of priapism was identified in this group. Early pharmacologic intervention for priapism induced by dynamic infusion cavernosometry cavernosography is a simple, safe, and time-saving measure to achieve detumescence and prevent potential sequelae such as corporal ischemia or fibrosis. PMID- 8235781 TI - Sensitivity and positive predictive value of clinical signs of hypogonadism in elderly men. AB - One hundred three ambulatory elderly men had serum free testosterone (FT) assessed as part of a study of bone loss. The FT was analyzed using radioimmunoassay. Each participant was questioned regarding the presence of erections adequate for sexual activity and the presence of sexual desire. Each was examined for decreased axillary and pubic hair. A FT of less than 9.0 pg/mL was found in eight subjects. The sensitivity of the clinical predictors as an indicator of a low FT value ranged from 43% to 86%, while positive predictive value ranged from 12% to 19%. The abnormal clinical signs and symptoms investigated in this study obviously have mechanisms in addition to hypogonadism. A history of adequate erections ruled out a low FT value in all but 1 of 44 cases. PMID- 8235782 TI - Serious adverse effects induced by simultaneous administration of two nonsteroidal anti-inflammatory drugs. AB - We have described the case of a 58-year-old woman in whom acute hemolytic anemia, renal failure, and granulocytopenia developed after intake of mefenamic acid and diclofenac. Results of direct and indirect Coombs' tests were negative, but the lymphocyte transformation test was positive for these two nonsteroidal antiinflammatory drugs (NSAIDs). We suggest that the immune complex mechanism could have induced these adverse reactions. Although these drugs are very popular, simultaneous administration should be avoided because serious adverse effects can potentially occur. PMID- 8235783 TI - Life-threatening hyperkalemia associated with captopril administration. AB - Angiotensin converting enzyme (ACE) inhibitors have become commonly used medications for hypertension and congestive heart failure. These agents are noted for their low incidence of adverse effects; but in certain cases, these effects can be life-threatening. Severe hyperkalemia is one of the potentially dangerous effects of the ACE inhibitors. While cases of life-threatening hyperkalemia associated with the use of ACE inhibitors have been described previously, in no instance was dialysis required. Herein, we report a case of acute hyperkalemia in a patient with congestive heart failure and renal insufficiency, the resolution of which required hemodialysis. The hyperkalemia in this case occurred without an increase in the patient's azotemia. In addition, the patient did not respond to attempts to effect the intracellular shift of potassium. This suggested that there may have been a defect in internal potassium homeostasis. PMID- 8235784 TI - Hypoglycemia due to a pharmacy dispensing error. AB - Drug dispensing error should be considered as a cause of hypoglycemia when the usual initial workup is unrevealing, as in the case described. PMID- 8235785 TI - Pure red cell aplasia and hepatitis A. AB - A patient with pure red cell aplasia associated with hepatitis A showed a dramatic and rapid recovery after initiation of oral prednisone therapy. The response to corticosteroid therapy suggests an immune etiology for this life threatening disorder. PMID- 8235786 TI - Use of thioridazine in post-traumatic stress disorder. AB - Post-traumatic stress disorder is a condition that develops in persons who have experienced emotional or physical stress of sufficient magnitude to be extremely traumatic for virtually anyone. This may include natural catastrophes, combat experiences, rape, or other such horrifying events. The three major features of the disorder are reexperiencing the trauma through dreams, emotional numbing, and autonomic instability. To date, several treatment modalities have been used, usually consisting of a combination of psychotherapy and drug treatment. Although controversy exists, antidepressants and monoamine oxidase inhibitors are used most commonly, while other drugs such as lithium, carbamazepine, and antipsychotic drugs may be useful. We have reported a case involving a 44-year old combat veteran who experienced severe flashbacks of his time spent in Vietnam. His symptoms and general state of mind improved significantly while taking the antipsychotic drug thioridazine. PMID- 8235787 TI - Rhabdomyolysis-induced delayed hypocalcemia after parathyroidectomy. PMID- 8235788 TI - Primary bronchopulmonary fibrosarcoma of the trachea in a child. AB - We have reported the unusual case of a 7-year-old boy who was admitted with respiratory symptoms of several months' duration. He was found to have a tumor of the trachea, which proved to be a low-grade fibrosarcoma with smooth muscle differentiation. PMID- 8235789 TI - Internal closure of an aspergilloma cavity: report of a new technique and review of the literature. AB - We have described a case in which closure of an aspergilloma cavity was done internally when it was determined that pulmonary resection was too hazardous. There was an intense inflammatory reaction surrounding the aorta and pulmonary artery. In the absence of gross purulence and underlying parenchymal infection, this technique may be used to avoid the prolonged hospitalization and bronchopleural fistulas associated with external drainage procedures. PMID- 8235790 TI - Neuroimaging of arachnoiditis induced by spinal anesthesia. AB - Despite the simplicity, effectiveness, and safety of spinal anesthesia, arachnoiditis can occur as a complication. The cause of this is often unknown, especially since disposable spinal kits are now normally used. It is postulated that an untoward reaction to the anesthesia or chemical contamination was the cause of arachnoiditis in our patient. We also outline the use of MRI in the diagnosis of arachnoiditis. This mode of investigation is often sufficient to establish the diagnosis. When necessary, it can be augmented by myelography and CT. PMID- 8235791 TI - Magnetic resonance imaging of nalbuphine-induced myocutaneous sclerosis. AB - Myocutaneous sclerosis is a known complication of intramuscular and subcutaneous injection of narcotics. We have described the MRI findings of this entity with superimposed infection in a patient addicted to intramuscular and subcutaneously administered nalbuphine. The clinical and imaging features of this disorder are sufficiently characteristic to allow confident diagnosis. This is the first report of this disorder due to nalbuphine. PMID- 8235792 TI - Metastatic choriocarcinoma: correlation of MRI, CT, and angiography. AB - We have described a 46-year-old woman with metastatic choriocarcinoma, anemia, and a liver mass. She subsequently had widespread metastatic disease and died despite chemotherapy. Without an antecedent history of pregnancy, her clinical presentation posed a diagnostic dilemma. This case emphasizes the importance of including choriocarcinoma in the differential diagnosis of liver masses in women of childbearing age and of correlating CT, MRI, and angiographic findings. PMID- 8235793 TI - Cystic pheochromocytoma: radiologic diagnosis. AB - Cystic pheochromocytomas are unusual variants of adrenal pheochromocytomas. Computed tomography and pathologic findings in the two cases reported here illustrate three cardinal principles in the diagnosis of cystic pheochromocytomas: (1) A suprarenal cystic tumor discovered on CT may be the result of hemorrhage and necrosis of an adrenal pheochromocytoma. (2) The cystic pheochromocytoma typically exhibits areas of low attenuation, with Hounsfield units in the range of 5 to 15 and rim enhancement on contrast administration. (3) Incidental adrenal lesions with these features discovered on CT scanning are an indication for catecholamine assays to screen for pheochromocytoma. PMID- 8235794 TI - Oropharyngeal impalement on a wrought iron fence. AB - Oropharyngeal impalement is a potentially life-threatening injury with protean manifestations. Attention to airway obstruction and active hemorrhage take initial priority. Foreign bodies are not removed outside the operating room unless they are causing immediately life-threatening compromise. The majority of injuries that cause thrombosis of the internal carotid artery are in the peritonsillar region of the lateral palate. These patients typically have a delayed onset of symptoms, sometimes more than 24 hours, so close observation is warranted. PMID- 8235795 TI - Systemic lupus erythematosus with retroperitoneal fibrosis and thrombosis of the inferior vena cava. AB - I have described a 17-year-old adolescent in whom retroperitoneal fibrosis was the initial manifestation of systemic lupus erythematosus. She had thrombosis of the inferior vena cava during the course of her illness. Standard treatment with corticosteroids resulted in apparent resolution of this complication. PMID- 8235796 TI - Cutaneous larva migrans. AB - The case of cutaneous larva migrans presented here is typical for its mechanism and geographic location of infection, evolution of lesions, and prompt response to treatment. Except for pinworms, helminth infections are rarely thought of in emergency departments away from the areas where the parasites are especially prevalent. The several-day incubation period and modern-day ease of travel should place this illness on one's list of the differential diagnoses of pruritic lesions regardless of the location of practice. This case serves as a reminder that in a mobile society, diseases, as well as patients, can travel. PMID- 8235798 TI - The winds of change in medicine. PMID- 8235797 TI - Delayed recurrences in primary fallopian tube carcinoma. PMID- 8235799 TI - Simultaneous nocardial empyema and Pneumocystis carinii pneumonia as an initial manifestation of HIV infection. PMID- 8235800 TI - WHO's work on the epidemiology of mental disorders. AB - The WHO programme on epidemiology of mental disorders started in the early 1960's with a series of reviews of knowledge. These were followed by activities aiming at four main goals: the standardization of psychiatric diagnosis, classification and statistics; the development of standardized internationally applicable instruments for the assessment of mental patients and of variables relevant to the assessment of mental illness; the conduct of epidemiological studies of mental disorders; and the training relevant to the above goals. The paper provides a description of the activities undertaken to achieve these goals and stresses the importance and usefulness of the collaboration of individuals, experts and institutions all over the world in this effort. PMID- 8235801 TI - Concordance between two measures of depression in the Hispanic Health and Nutrition Examination Survey. AB - A concordance analysis between the Center for Epidemiologic Studies Depression Scale (CES-D) and the National Institute of Mental Health (NIMH) Diagnostic Interview Schedule (DIS) for current major depression was conducted using data from Cuban Americans and Puerto Rican respondents to the Hispanic Health and Nutrition Examination Survey (HHANES). Overall agreement between the two depression measures was relatively high, which suggested that the CES-D might be appropriate as a first-stage screening instrument for community-based surveys of clinical depression. Female gender and indicators of social class (education, income, poverty index, and employment) were related to low specificity and low agreement. The estimated cutoff points of the CES-D that best predicted DIS current major depression were different between the two ethnic groups; 17 for Cuban Americans, and 20 for Puerto Ricans. A receiver operating characteristics (ROC) curve analysis revealed that the traditional method of defining CES-D cases by summing the scores for each item was superior to counting only the persistent symptoms, that is, those present nearly everyday. PMID- 8235802 TI - Subtypes in schizophrenic disorders: frequencies in long-term course and premorbid features. AB - The subclassification of schizophrenic disorders according to four diagnostic systems (DSM-III-R, ICD-10, the positive vs. negative dichotomy and Schneider's first rank symptoms) was compared over the long-term course of the disease in 148 narrowly defined schizophrenic patients. A total of 595 episodes were classified over a mean observation period of 23 years (range 10-50 years). Initially, paranoid/positive subtypes predominated, while later in the course episodes fulfilling the symptomatological criteria of residual/negative subtypes became more frequent. Disorganised/hebephrenic and catatonic subtypes were found to be rare. Some premorbid features were investigated as non-symptomatological validators for subclassification. Significant differences were found with regard to age at onset. Patients whose first episode was paranoid or positive had the highest age at onset. Patients with initial disorganised/hebephrenic or "residual" episodes had the most unfavourable premorbid social adjustment, even when the influence of age at onset was discounted. The diagnostic systems investigated showed similarities and differences as a result of the underlying concepts. Methodological implications are discussed. PMID- 8235803 TI - The pattern of burden in families of neurotic patients. AB - The pattern of burden in 90 families with a neurotic patient, 30 each with dysthymia, generalized anxiety disorder and obsessive compulsive disorder was evaluated in this study. Burden was chiefly felt in four principal areas; financial burden, disruption of family routine, disruption of family leisure and family interactions. This pattern was similar among the three study groups. Analysis of problems perceived as burdensome demonstrated an increased concern among relatives regarding routine household matters, restricted leisure and ill effects of the patient's illness on the family atmosphere. Burden was significantly greater in cases of married patients, married relatives, housewives and employed patients. Such data, it is hoped, will help provide guidelines along which an enquiry into different aspects of burden can be conducted. PMID- 8235804 TI - Social functioning as a significant factor in women's help-seeking behaviour during the climacteric period. AB - In order to identify the psychosocial factors that lead to the demand for medical care related to the menopause, we carried out a case/control study. A case was defined as a woman who sought gynaecological care due to "menopausal complaints" (n = 85) and a control referred to a woman drawn at random from the general population. The cases showed greater psychiatric morbidity and social dissatisfaction, a lower level of diffused social support and a higher frequency of severe life events and the controls showed greater social maladjustment in objective conditions. The multivariate analysis (logistic regression) carried out after adjusting all the relevant variables, indicated that the demand for medical care due to menopausal complaints was not only determined by the menopausal status itself but also by a set of social and demographic variables, especially dissatisfaction and a lack of social support. PMID- 8235805 TI - A weighted time budget approach for the assessment of cognitive and social activities. AB - An instrument designed to assess potentially challenging cognitive and social activities in a wide range of both psychiatric and normal study populations is described in this paper. The technique used combines traditional time budget approaches with an additional, significant component, i.e. a rating indicating the amount of challenge for each activity that is sampled. Separate ratings reflecting the subject's and the interviewer's perspective are generated. Along with a detailed description of this weighted time budget instrument, preliminary data regarding practicality, reliability and validity are reported. An analysis based on time budget interviews with 30 schizophrenic patients showed overall interrater agreement to average 84% (kappa = 0.72). In a study with a possible bearing on the concurrent validity of the instrument, relapsing schizophrenics significantly differed from stable subjects in weighted time budget indices. PMID- 8235806 TI - Perception of problems in psychiatric inpatients: denial, race and service usage. AB - The importance of the ways in which people with psychiatric problems construe their difficulties is considered in this study. A study of 60 consecutive acute admissions to wards serving an inner city area in London (UK) is reported. The results indicated that 55.8% of the sample did not consider themselves to have psychiatric problems: 15.4% said that they had no problems at all and 40.4% thought they had physical or social problems rather than psychiatric ones. Although more younger people denied that they had problems and none of those who denied having problems sought the help of a general practitioner, there was a significant association between diagnosis and perception of problems, and when this was taken into account these associations disappeared. Of those who denied having any problems, only one person had no police involvement on admission. Significantly more of those who denied problems were compulsorily admitted and there were significant differences in the proportions of whites and African Caribbeans reporting different types of problems. African-Caribbeans were both more likely to consider that they had no problems at all and to be compulsorily admitted. Although African-Caribbeans were also more likely to be diagnosed as experiencing psychotic disorders, it was their ethnic status rather than their diagnostic category that determined both their status on admission and the way in which they construed their problems. Denial among whites tended to take the form of somatisation or construction of problems in terms of social difficulties. PMID- 8235807 TI - Post-traumatic stress disorder and coping after a natural disaster. AB - This study examines the role of coping in the onset of post-traumatic stress disorder (PTSD) in a nonpatient population following exposure to a natural disaster. In contrast to other studies, the use of all coping strategies was found to be associated with the presence of PTSD rather than the absence of symptoms. These data suggest that coping (in this sense) represents a psychological process used to contain the distress caused by symptoms as well as to manage environmental adversity. PMID- 8235808 TI - Evaluation of an adaptive behaviour classification for mentally retarded adults. AB - Adaptive behaviour was assessed in a total of 119 mentally retarded adults by means of the Disability Assessment Schedule (DAS). According to a newly developed classification scheme, the level of handicap was graded on a four-step scale according to the subject's score in adaptive behaviour on the DAS; this was highly correlated both to the Coloured Progressive Matrices (CPM)-IQ (r = 0.75) and the Columbia Mental Maturity (CMM) Scale (r = 0.77). The DAS classification was associated with relevant neuropsychiatric variables. The findings indicate that the DAS classification represents a practicable way for assessing the degree of handicap in mentally retarded adults and could be especially valuable in cases where intelligence testing proves impossible, as was the case for 45% (CPM) and 39% (CMM) of the present sample. PMID- 8235809 TI - A randomized, placebo-controlled trial of exercise therapy in patients with acute low back pain. AB - To assess the efficacy of exercise therapy for acute low back pain, a randomized, placebo-controlled trial was performed in 40 Dutch general practices. Patients received either exercise instruction with advice for daily life by a physiotherapist; placebo ultrasound therapy by a physiotherapist; or usual care by the general practitioner. All patients received analgesic agents and information on low back pain before randomization. Four hundred seventy-three patients were included. No differences in number of recurrences, functional health status, or medical care usage could be found among the three groups. In the exercise group, duration of recurrences was shorter and patients were less tired during the first 3 months than in the usual care group, but no differences were found between the exercise and placebo groups. It was concluded that exercise therapy for patients with acute low back pain has no advantage over usual care from the general practitioner. PMID- 8235810 TI - A stochastic model of trunk muscle coactivation during trunk bending. AB - Biomechanical models of the spine have traditionally assumed that workplace lifting conditions (weight, posture, motion, etc.) precisely dictate the magnitude of individual muscle forces necessary to maintain a biomechanical balance within the trunk. However, because there are a large number of muscle groups within the trunk there is also an infinite number of possible combinations of muscle forces that can satisfy this biomechanical balance requirement for a given condition. Currently there are no methods available to predict this possible variability in muscle activity. Such variability in a multiple muscle system can result in variations in spinal loading. To quantitatively capture this trunk muscle variability during bending motions, such as those involved in lifting, a stochastic (probabilistic) model of trunk muscle activation was developed. The model was based on a simulation of experimentally derived data and predicted the possible combinations of time-dependent trunk muscle coactivations that could be expected given a set of trunk bending conditions. These simulated muscle activities were then used as input to an electromyographically assisted biomechanical model so that the magnitude and variability of the spine reaction forces could be estimated. This procedure allows one to assess the range of spinal loads that would be expected with a particular task. Significant variability in muscle activities was observed for each specific lifting condition and explained biomechanically. The results indicated that the variability in trunk muscle force had a small effect on spinal compression variability (+/- 7% of the mean compression), but greatly influenced both lateral (+/- 90% of mean) and anteroposterior shear forces (+/- 40% of mean). A validation study confirmed that the model predictions were reasonable estimates of muscle activity variability under previously untested conditions. This work could help explain how some repetitive lifting motions could increase the risk of acquiring a low back disorder and the simulation model could help drive electromyographically assisted models without the need for recording actual electromyographic activity. PMID- 8235811 TI - Vasogenic edema induced by compression injury to the spinal nerve root. Distribution of intravenously injected protein tracers and gadolinium-enhanced magnetic resonance imaging. AB - The function of the blood-nerve barrier appears quite unique in the nerve root. Protein tracers that were injected into the subarachnoid space passed through the nerve root sheath and entered into the capillary lumen in the endoneurial space but tracers that were injected intravenously did not appear in the endoneurial space. Marked extravasation of protein tracers in the nerve root was induced at the compressed part by strong compression (60 gram force, 30 gram force) and capillaries in the nerve root showed opening of the tight junction accompanied by an increase in vesicular transport under the electron microscope. This situation was reflected as high intensity on Gadolinium-enhanced magnetic resonance imaging. In twenty-one of fifty patients with lumbar disc herniation, the affected nerve root was strongly enhanced by Gadolinium-diethylene triaminepentaacetic acid, indicating that the blood-nerve barrier in the affected nerve root was broken and intraradicular edema was produced in these cases. PMID- 8235812 TI - Autologous nucleus pulposus induces neurophysiologic and histologic changes in porcine cauda equina nerve roots. AB - Epidural application of autologous nucleus pulposus in pigs, without mechanical nerve root compression, induced a pronounced reduction in nerve conduction velocity in the cauda equina nerve roots after 1-7 days, compared to epidural application of retroperitoneal fat in control experiments. Histologically, the nerve fiber injury was more pronounced after application of nucleus pulposus than after control tissue application. The results demonstrate that nucleus pulposus may induce nerve tissue injury by mechanisms other than mechanical compression. Such mechanisms may be based on direct biochemical effects of nucleus pulposus components on nerve fiber structure and function and microvascular changes including inflammatory reactions in the nerve roots. PMID- 8235813 TI - The natural course of acute sciatica with nerve root symptoms in a double-blind placebo-controlled trial evaluating the effect of piroxicam. AB - To study the natural history of acute sciatica, 208 patients with obvious symptoms and signs of a lumbar radiculopathy (L5 and S1) were examined within 14 days of onset. A concomitant double-blind investigation of the effect of the nonsteroidal anti-inflammatory drug piroxicam was performed. The results measured by visual analog scale and Roland's functional tests showed a satisfactory improvement throughout the 4 weeks of observation. The piroxicam-treated group had same results as the control group. Based on questionnaires at months 3 and 12 approximately 30% of the patients still complained about back trouble and 19.5% were out of work after 1 year. Four patients underwent surgery during this period. PMID- 8235814 TI - Muscle fiber distribution, capillary density, and enzymatic activities in the lumbar paravertebral muscles of young men. Significance for isometric endurance. AB - The aim of this work is to expand the knowledge about fiber distribution, capillarization, and metabolic potential in different anatomic sections of the left and right human lumbar paravertebral muscles and further to investigate whether a relationship between these parameters and trunk muscle endurance exists. Lumbar (L3) paravertebral muscle samples were taken bilaterally from six young male cadavers (study 1) and from 10 normal young healthy men (study 2). The main findings are that the paravertebral muscles are dominated by slow twitch fibers with small fiber areas and a well-developed capillary network. This is most pronounced in the longissimus muscle. Glycogen concentration and enzyme activities (glycogen phosphorylase, lactate dehydrogenase, citric synthetase and 3-hydroxyacyl-CoA-dehydrogenase) in both aerobic and anaerobic metabolic pathways are large compared to other muscles. No obvious bilateral differences in the histochemical and biochemical results were present. The isometrical trunk extensor endurance time is markedly increased compared to other human skeletal muscles. PMID- 8235815 TI - Gait analysis of patients with low back pain before and after surgery. AB - Gait analysis could be a good objective and quantifiable assessment of function in patients with chronic low back pain. A study of the gait parameters of 20 normal subjects and 30 patients with low back pain, before and 2 years after surgery (mainly spinal arthrodesis) was undertaken. For the majority (67%) of patients, postoperative gait analysis confirmed their perception of pain and function. In a minority (33%) of patients, there was an improvement in gait parameters, despite the perception of continuing severe pain and poor physical functioning. It may be possible to reduce the number of patients experiencing symptomatic failure with surgery if this discrepancy between gait and disability due to pain can be discovered preoperatively. Then the advice against surgery could be given when indicated. PMID- 8235816 TI - A pathologic study of discs in the elderly. Separation between the cartilaginous endplate and the vertebral body. AB - Eighty-eight autopsy specimens from L4-5 lumbar discs of cadavers with an average age of 77.6 years were examined radiologically and histopathologically. They were classified into three groups by the height of intervertebral disc space: slightly degenerated (57 cases), moderately degenerated (25 cases), and severely degenerated (6 cases). Cartilaginous end-plate rupture was found most commonly in the severely degenerated group. Reverse orientation of the anulus fibrosus was found in one third of all specimens. End-plate was separated from vertebral body in 45 cases (51.1%) of 88 specimens. From the findings it is suggested that the end-plate is avulsed from the vertebral body under the precondition of separation and then herniated with anchoring anulus fibrosus. This type of herniation occurs more often than herniation of the nucleus pulposus in the elderly. PMID- 8235817 TI - Lumbar spinal fusion. A cohort study of complications, reoperations, and resource use in the Medicare population. AB - Regional variations in lumbar spinal fusion rates suggest a poor consensus on surgical indications. Therefore, complications, costs, and reoperation rates were compared for elderly patients undergoing surgery with or without spinal fusion. Subjects were Medicare recipients who underwent surgery in 1985, with 4 years of subsequent follow-up. There were 27,111 eligible patients, of whom 5.6% had fusions. Mean age was 72 years. Patients undergoing fusion had a complication rate 1.9 times greater than those who had surgery without fusion. The blood transfusion rate was 5.8 times greater, nursing home placement rate 2.2 times greater, and hospital charges 1.5 times higher (all P < 0.0005). Six-week mortality was 2.0 times greater for patients undergoing fusions (P = 0.025). Reoperation rates at 4 years were no lower for patients who had fusion surgery and results were similar in most diagnostic subgroups. Indications for fusion among older patients require better definition, preferably based on outcomes from prospective controlled studies. PMID- 8235818 TI - Long-term results of surgical treatment of lumbar spinal stenosis. AB - There are few data available regarding the long-term outcome of laminectomy for lumbar canal stenosis. Both in 1985 and in 1991 108 patients who had undergone surgery for lumbar spinal stenosis were reevaluated using the Oswestry disability questionnaire. There were 50 women and 58 men. The clinical diagnosis of stenosis was confirmed mainly by myelography. The mean follow-up time in 1985 was 6.8 years and in 1991 12.8 years, and the mean age of the patients at the time of surgery was 50.7 years. The mean Oswestry score in 1985 was 34.5 and in 1991 30.2 (P < 0.001). Men improved more than women. Laminectomy was performed on an average of 1.6 levels. Ten (9.3%) of the 108 patients underwent repeat decompression during the study period. The authors concluded that the results of the patients improved during the course of the longitudinal follow-up time of 7 and 13 years. The chances of a patient requiring reoperation after surgery for lumbar spinal stenosis were low. PMID- 8235819 TI - Decompression for degenerative spondylolisthesis and spinal stenosis at L4-5. The effects on facet joint morphology. AB - Anatomic variations exist in the facet joint orientation, shape, and size at L4 5. This morphology is further modified by degenerative changes in spinal stenosis and degenerative spondylolisthesis. This study explored the morphologic alteration of "pedicle-to-pedicle" decompression on the facet joints in normal patients, spinal stenosis patients, and degenerative spondylolisthesis patients. Using computerized digitization, computed tomographic scan images of the facet joint at L4-5 and the medial border of the pedicle at L5 were superimposed. The facet joint orientation, coronal dimension, percentage, and absolute reduction in coronal dimension after pedicle-to-pedicle decompression, and residual coronal dimension after decompression at L4-5 were measured for the three groups. There is a significantly reduced coronal dimension of the facet joint in degenerative spondylolisthesis patients compared with spinal stenosis and normal patients (P < 0.01). The average reduction of the facet joint coronal dimension is 34% (SD 30%) in degenerative spondylolisthesis, and 36% (SD 25%) in spinal stenosis. The smaller preoperative coronal dimension in degenerative spondylolisthesis leads to a significantly reduced residual coronal dimension in degenerative spondylolisthesis compared with normal patients (5.9 mm [SD 4.3 mm] vs. 9.3 mm [SD 3.5 mm]), respectively. Wide variations in facet joint reduction and residual facet joint coronal dimension exist. The significantly reduced coronal dimension after decompression in degenerative spondylolisthesis may be correlated to a trend to further anterior displacement if it is treated with decompression alone. Case-specific assessment of residual facet joint morphology after decompression in both spinal stenosis and degenerative spondylolisthesis patients should be integrated into decisions about fusion for stability at the L4-5 level. PMID- 8235820 TI - Postoperative spondylolisthesis at L4-5. The role of facet joint morphology. AB - Thirty-three patients underwent decompression without fusion at the L4-5 level for spinal stenosis or degenerative spondylolisthesis. Using preoperative and 1 year postoperative lateral lumbar spine radiographs, the incidence of postoperative spondylolisthesis of greater than 5% was found to be 58%. Computed tomographic scans were used to analyze the presurgical facet joint morphology and facet joint-pedicle spatial relationship. This allowed calculation of the facet joint orientation for each side; the coronal dimension of each facet joint; the amount of the facet joint coronal dimension removed if a decompression was performed up to the medial border of the L5 pedicle (facet joint reduction); and the residual coronal dimension of facet joint after such a decompression (residual facet joint). The lateral radiographs were analyzed for presurgical disc height and the presence of traction spurs or spondylophytes. A well maintained disc height was associated with an increase slip (7.47%) compared with those cases with a narrow or complete loss of disc space before surgery (4.84% P < 0.1 trend). Presence of spondylophytes was associated with a reduced tendency to slip. When spondylophytes were controlled for there was a significant relationship between slip of greater than 10% and sagittal facet joint orientation. Although there was a lesser residual facet joint after decompression in the group that slipped these values were not statistically significant. This study suggests that the development of postoperative spondylolisthesis is related to facet joint orientation and dimensions, rather than the absolute amount of joint removed. The stabilizing effects of reduced disc height and spondylophytes were confirmed. PMID- 8235821 TI - Failure of a modified posterior lumbar interbody fusion technique to produce adequate pain relief in isthmic spondylolytic grade 1 spondylolisthesis patients. A prospective study of 20 patients. AB - A modified posterior lumber interbody fusion procedure is described: this technique basically uses only two tricortical bone grafts and does not produce a distraction at the interspace while inserting the bone grafts. Twenty patients with intractable back and leg pain due to an isthmic spondylolysis and a grade 1 spondylolisthesis were operated using this technique and a prospective study with a mean follow-up of 41 months was conducted to evaluate the results. The final evaluation involved a questionnaire, a clinical examination, and a lumbar spine radiographic series. Fourty-five percent of the patients had good or excellent results whereas 55% had fair or poor results. The possible reasons for these unfavorable results are discussed. PMID- 8235822 TI - The association of mild-moderate isthmic lumbar spondylolisthesis and low back pain in middle-aged patients is weak and it only occurs in women. AB - This article reports on a cohort-like study of 20 women and 26 men with spondylolisthesis derived from a population of 1147 randomly selected 45-64-year old subjects against sex-matched control subjects derived from the same source. Seventy-five percent of the spondylolisthetic subjects belonged to Meyerding class I representing mild slip. Low back symptoms during the previous day were more prevalent in women with spondylolisthesis than in control subjects (risk ratio was 7.0; 95% confidence interval 1.4-35.7), and the back pain index was higher (P = 0.028). No significant differences were seen between the groups of men in back symptoms. The use of healthcare facilities and the proportion of subjects receiving invalidity pension because of low back disease did not show any differences between the study groups of either sex. In subjects with spondylolisthesis the degree of vertebral body slip was associated with degeneration of the olisthetic intervertebral disc (P = 0.003), but it was not associated with increased occurrence of back symptoms or pain index. These data suggest that mild-moderate spondylolisthesis detected by chance in a middle-aged population does not predispose to more disabling back pain than back problems experienced by those without spondylolisthesis. However, women with spondylolisthesis have mild back symptoms more often than control subjects, suggesting that in clinical practice and future studies the sex of the patient should be considered in the evaluation of the importance of radiologic spondylolisthesis. PMID- 8235823 TI - Spinal shrinkage as a parameter of functional load. AB - This article reviews the possibilities of spinal shrinkage in ergonomic evaluation of working situations. Because long-term health effects due to spinal loading are difficult to assess, short-term effects of spinal loading, such as spinal shrinkage, are used to obtain insight in the workload and its consequences, based on an assumed predictive value regarding health effects. The mechanism underlying spinal shrinkage (i.e., loss of motion segment height) and its possible relationship to low back pain, the way in which spinal shrinkage has been measured in vivo, and some results of in vivo studies are discussed. Conclusions are drawn on the validity and applicability of spinal shrinkage in ergonomic evaluations. The validity needs further investigation in relation to the loads imposed on the spine, the capacity of the worker, and the health consequences. PMID- 8235824 TI - Pivoting with the load. An alternative for protecting the back in asymmetrical lifting. AB - The purpose of this study was to examine different lifting conditions with combined flexion and twisting of the trunk relative to the pelvis (full twist and half-twist techniques) and to compare them to a situation using feet pivoting with minimization of trunk motions relative to the pelvis. Nine subjects with limited experience were tested with two force platforms and two 16-mm cameras coupled with two mirrors; dynamic three-dimensional multisegment models were constructed and the net muscular moments as well as the angular velocities of the trunk relative to pelvis were determined about the three orthogonal orthopaedic axes of the trunk at L5-S1 in flexion-extension, lateral bending, and twisting. The results showed that pivoting required larger extension moments but considerably smaller twisting moments and that these moments were exerted from a safer trunk posture. Pivoting appears to be a good compromise to avoid or minimize complex trunk postures and loadings. PMID- 8235825 TI - Can intervertebral disc prolapse be predicted by disc mechanics? AB - The hypothesis was tested that stress concentrations in the posterior anulus of an intervertebral disc predispose it to prolapse under high compressive loads and anterolateral bending. The distribution of compressive stress inside the intervertebral discs of 22 cadaveric lumbar motion segments was measured with the specimens loaded in pure compression and in compression combined with anterolateral bending. Each motion segment was then loaded to failure in combined compression and anterolateral bending. Failure occurred in the vertebral body (n = 12) or posterolateral anulus (n = 10); the latter group showed a significantly greater incidence of stress concentrations (P < 0.001) in the posterior anulus, when loaded in compression and bending. It was concluded that some discs are predisposed to prolapse because of damaging, localized concentrations of stress in the posterior anulus in combined anterolateral bending and compression. PMID- 8235826 TI - A combined finite element and optimization investigation of lumbar spine mechanics with and without muscles. AB - A combined finite element and optimization approach to study the effects of muscles on the biomechanics of the lumbar spine was initiated. Briefly, a three dimensional, nonlinear, finite element model of a ligamentous L3-4 motion segment was formulated (LIG model) for the predictions of stresses, etc., in the motion segment. A separate, biomechanical optimization-based force model with experimental input was developed to predict the forces in muscles and disc across the L3-4 segment in response to a person holding 90 N in his hands with spine flexed 30 degrees, and knees straight. The predicted muscle forces from the optimization model were then incorporated into the L3-4 finite element model as nodal forces to simulate the muscle action (MUS model). The predicted responses from the muscles active (MUS) finite element model were compared to the corresponding results from the ligamentous (LIG) finite element model subjected to an equivalent load. The biomechanical parameters compared were: translation and rotation of L3, disc bulge, intervertebral foramen gap, intradiscal pressure, facet loading, ligament tension, compressive disc load, and stresses in the vertebral body. The addition of muscular forces in the MUS model led to a decrease in the anteroposterior translation and flexion rotation (displacements in the sagittal plane) of the segment compared to the corresponding LIG model predictions. Thus, the muscles imparted stability to the ligamentous segment. The presence of muscles also led to a decrease in stresses in the vertebral body, the intradiscal pressure and other mechanical parameters of importance. However, the load bearing of the facets increased compared to the ligamentous model. Thus, facets play a significant role in transmitting loads in a normal intact spine. These results, for the first time, provide quantitative data on the stabilizing effects of muscles on the mechanics of a ligamentous spine. The results also provide a scientific explanation in support of the "degenerative cascade" concept proposed in the literature. The model predictions, in conjunction with the degenerative cascade concept, also support the observation that the osteoarthritis of facets may follow disc degeneration. Future research directions based on the current model are presented. PMID- 8235827 TI - The pial ligaments of the anterior spinal artery and their stretch receptors. A spinal cord distraction-sensing system? AB - The pial sheath of the anterior spinal artery displays a system of ligaments that course along the ventral surfaces of the anterior spinal artery and its medullary feeder arteries on the lower half of the spinal cord. Frequently, discrete ligamentous straps extend from these anterior spinal artery ligaments to the sheath of an anterior spinal nerve root to reinforce the general cauda equina pial connections to this system. Microscopy of ligament sections revealed that numerous Golgi-type neurofascicular receptors were oriented longitudinally among the ligament fascicles and associated with well-myelinated nerves. As this type of mechanoreceptor has been known only in association with stretch reflex mediation in the musculoskeletal system, it appeared likely that these anterior spinal artery ligaments and their homologous type of receptors may be implicated in sensing distraction of the thoracolumbar spinal cord and protectively modifying the actions of the involved spinal musculature. PMID- 8235828 TI - The effects of induced hypertension and acute graded compression on impulse propagation in the spinal nerve roots of the pig. AB - Injuries caused by compression of spinal nerve roots are frequently encountered clinically. Experimental studies show that several different factors affect the pathophysiologic changes that occur after these injuries. However, the effect of hypertension in conjunction with graded compression of spinal nerve roots is yet unclear. A previously established porcine model was employed, in which the spinal nerve roots were exposed and compressed by an inflatable balloon. Impulse propagation across the compressed nerve segment was studied by the recording of efferent and afferent nerve action potentials, and nerve conduction velocity. The systemic blood pressure was increased by administration of Neo-Synephrine hydrochloride (phenylephrine HCl) (Winthrop Pharmaceuticals, New York, NY) and elevated 40 +/- 5 mm Hg above the normal (100 +/- 5 mm Hg), and electrophysiologic baseline values were recorded. The spinal nerve roots were then compressed for 2 hours with either 0 (control), 50, 100, or 200 mm Hg. The balloon was deflated and the nerve roots were allowed to recover for 1.5 hours. Impulse propagation was studied every 15 minutes and hypertension was maintained throughout the experiment. The results showed no significant changes of the efferent and afferent nerve action potentials at 0.50 or 100 mm Hg. At 200 mm Hg, efferent and afferent nerve action potentials decreased rapidly and were almost abolished within 20 minutes of compression. Some but not significant recovery was seen of the nerve impulse. Compared to data from normotensive pigs in an earlier experiment, the current study showed that hypertension significantly decreases the susceptibility of the spinal nerve roots to compression at and below 100 mm Hg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235829 TI - Does Harrington instrumentation improve pulmonary function in adolescents with idiopathic scoliosis? A meta-analysis. AB - A meta-analysis of the effects of Harrington instrumentation on vital capacity in adolescent patients with idiopathic scoliosis was conducted in an attempt to clarify the conflicting conclusions of different studies. Thirty-eight studies were assessed. Those that did not prevent sufficient statistical information and those that did not allow for normal growth during the interval between tests were excluded. The five studies remaining for analysis, involving a total of 173 patients, showed an increase in mean vital capacity ranging from 2% to 11% of predicted vital capacity. The weighted mean effect size for the five studies analyzed was 0.22 with 95% confidence intervals of 0.01-0.43. The authors conclude that Harrington instrumentation leads to a small but statistically significant improvement in vital capacity. PMID- 8235830 TI - Combination of acute normovolemic hemodilution and deliberate hypotensive anesthesia. PMID- 8235832 TI - Scoliosis Research Society, 26th annual meeting. Proceedings. Kansas City, Missouri, September 23-26, 1992. PMID- 8235831 TI - Conventional discectomy. PMID- 8235833 TI - Outcome of spinal screening. AB - Spinal screening using the Scoliometer was performed on one thousand high school students to determine the prevalence of spinal deformity and the appropriate degree of deformity to be used as a selection criterion. Eighty percent of students were found to have 3 degrees or more angle of trunk rotation (ATR). This new information predicts a referral rate of three percent if 7 degrees degrees ATR is used as referral criterion (12% referral if 5 degrees ATR is the criterion). A paradigm using the best available information on natural history and screening allows a comparison of the outcomes of screened versus non-screened school populations. Spinal screening appears to be effective in reducing the need for surgical treatment of scoliosis, but does not decrease the total cost of care for this problem. An appropriate referral criterion for spinal screening appears to be 7 degrees ATR at any level of the spine. PMID- 8235834 TI - Diurnal variation of Cobb angle measurement in adolescent idiopathic scoliosis. AB - To investigate the possible influence of gravity on daily variations of curve magnitude in adolescent idiopathic scoliosis, standardized anteroposterior standing radiographs of the spine were taken at 8:00 AM and repeated in the same conditions at 8:00 PM on the same day, 19 girls with idiopathic scoliosis, aged 10-16 years were included. The Cobb angle of the primary thoracic curve was measured blindly by four skilled orthopaedic surgeons. The mean interobserver variation in measuring Cobb angles on the 38 radiographs was 1.6 degrees. The average Cobb angle on morning radiographs was 60 degrees (range, 42-91 degrees) as compared to 65 degrees (range, 47-89 degrees) in the afternoon, a significant (P < 0.001) difference using a paired two-sided Student t test. It was concluded that there is a statistically and clinically significant daily increase of curve severity in moderate to severe idiopathic scoliosis. PMID- 8235835 TI - Outcome of Wisconsin instrumentation in idiopathic scoliosis. Minimum 5-year follow-up. AB - Wisconsin segmental spinal instrumentation was developed by Drummond et al to provide rigid fixation for correction of spinal deformities without the risk of neurologic injury associated with the passage of sublaminar wires. Up to this point, there have been no outcome studies with follow-up of longer than 2 years. Thirty-five patients with adolescent idiopathic scoliosis treated by the Wisconsin procedure between 1984 and 1987 were reviewed by radiographs, physical examination, and an outcome scale. Minimum length of follow-up was 5 years with an average of 6.3 years. Curves were corrected from a mean of 59 degrees preoperatively to 32 degrees postoperatively (46%), and 36 degrees at final follow-up (39%). Wisconsin segmental spinal instrumentation did not increase thoracic kyphosis. There was evidence of slight lumbar flattening in long fusions. According to the criteria described, 92% had a successful outcome. Complications of the procedure included two wound infections (one superficial, one deep), one rod displacement, and two wire breakages. No pseudarthroses or neurologic complications were identified in this series. Wisconsin segmental spinal instrumentation safely achieves the objectives of partial correction, arthrodesis, and early return to function. The technique may still have a role, particularly in the thoracic region, in selective thoracic fusion of King type II curves, rigid curves, and double thoracic curves. PMID- 8235836 TI - Scoliosis associated with syringomyelia. AB - From January 1986 to December 1991, 1,059 patients with scoliosis of 20 degrees or more were reviewed. By magnetic resonance imaging, 43 patients were found to have syringomyelia. Thirty-eight patients had Chiari type 1 malformation. Twenty two of the patients were younger than 10 years. Scoliosis was the only presenting symptom in 32 patients. All patients had some neurologic abnormalities. Fifteen patients had left thoracic curves. The incidence of scoliosis with syringomyelia in our outpatient clinic was 4.0%; 18.4% in boys, and 2.6% in girls. Magnetic resonance imaging should be performed in patients who have abnormal neurologic findings. PMID- 8235837 TI - Are there indications for partial rib resection in patients with adolescent idiopathic scoliosis treated with Cotrel-Dubousset instrumentation? AB - In adolescent idiopathic scoliosis the cosmetically unacceptable rib prominence is one of the main reasons patients seek treatment. Ninety-eight patients were reviewed with Cotrel-Dubousset (CD) instrumentation (average follow-up, 27 months; average preoperative curve, 53 degrees) and 27 patients with Harrington instrumentation and rib resection (average follow-up, 43 months; average preoperative curve, 52 degrees). Of the 98 patients who had CD instrumentation, 15 underwent concomitant rib resection. Of the 83 patients without rib resection, 71 (72%) were rated satisfactory and 12 unsatisfactory by cosmetic criteria based on residual rib deformity. These results were compared to those of 27 patients receiving Harrington rod instrumentation and rib resection, of whom 23 (85%) were rated as satisfactory. All 15 patients with CD and rib resection were rated satisfactory. The CD patients were then redivided into two groups (rib resection indicated or rib resection not indicated) as follows: the 12 unsatisfactory CD patients without rib resection (in whom a rib resection should have been done) were grouped with the 15 CD patients who underwent rib resection, for a total of 27 rib resections indicated, or 28%. This group was compared to the 71 satisfactory CD patients without rib resection (rib resection not indicated). Patients with a rib prominence of > 15 degrees preoperatively had or should have had a rib resection. Patients with a higher chance of needing rib resection included those with a curve severity greater than 60 degrees, curve flexibility less than 20%, a preoperative rib prominence > 10 degrees, or intraoperative curve correction of less than 50%. PMID- 8235838 TI - Analysis of the upper thoracic curve in surgically treated idiopathic scoliosis. A new concept of the double thoracic curve pattern. AB - The authors reviewed 246 idiopathic scoliosis patients with the upper thoracic curve of more than 20 degrees. Group I (138 patients) had positive T1 tilt and a spinal fusion which was extended over both the upper and lower thoracic curve with the diagnosis of double thoracic curve. Group II (43 patients) had positive T1 tilt, but the fusion was limited to the lower thoracic curve. Group III (65 patients) had negative or neutral T1 tilt and the fusion was limited to the lower thoracic curve. The average age at operation was 15.9 years (range, 11.2-35 years) and the average length of follow-up was 4.8 years (range, 2-29.5 years). Positive T1 tilt did not correlate well with left shoulder elevation contrary to previous reports. The upper thoracic curve was more rigid than the lower curve in all groups and the lumbar curve was significantly more flexible than the upper and lower thoracic curves in all groups (P < 0.05). No significant difference in the flexibility of the upper thoracic curve was found between the groups regardless of the direction of T1 tilt. When only the lower curve was fused (groups II and III), progression of the upper thoracic curve was less than 5 degrees, and spontaneous correction of the unfused upper curve occurred in the majority of the cases following the supine bending study. Correction and fusion on the lower curve (groups II and III) aggravated shoulder imbalance of all patients with left shoulder elevation. Based on the findings of this study, the authors proposed that the diagnosis of idiopathic double thoracic patterns should be limited to those patterns which require fusion of both the upper and lower curves. This pattern of idiopathic scoliosis includes double thoracic curves with left shoulder elevation and/or a rigid upper thoracic curve. PMID- 8235839 TI - An experimental study in chickens for the pathogenesis of idiopathic scoliosis. AB - Experimentally induced scoliosis was investigated in pinealectomized chickens using pathologic and neurophysiologic means. A total of 90 chickens were tested; 30 served as a normal control, 30 received an autografted pineal body in the intramuscular tissue of the trunk, and 30 underwent pinealectomy without autograft. Scoliosis developed in all pinealectomized chickens within 2 weeks, showing gradual progression during the next 5 or 6 weeks. At 3 months, the three dimensional spinal deformity consisted of lateral curvature and vertebral body rotation, resulting in a prominent lordoscoliosis at the thoracic level. In contrast, scoliosis developed in only 10% of the autografted chickens. Histologic examination revealed no pathologic change in the brain in either the pinealectomized scoliosis group or in the autografted nonscoliosis group. Cortical potentials in the scoliosis group were delayed, thus suggesting conduction disturbance rostral to the brain stem. Although the relationship between the cause and effect is uncertain, these findings implicate neurotransmitters or neurohormonal systems in the pineal body as a major contributing factor in this type of experimental scoliosis. PMID- 8235840 TI - Vertebrodiatasis experimental lengthening of the juvenile goat spine. AB - The purpose of this study is to describe the biology of spinal lengthening (vertebrodiatasis) in a juvenile goat model. A small Orthofix dynamic fixator was affixed to 5.5 mm pedicular halfpins placed bilaterally at T12, L1, L5 and L6 in six juvenile goats. Distraction was performed across vertebral endplates between L1 and L5. Uninstrumented twins served as controls. Two pairs of animals were deleted due to anesthetic complications. Lumbar lengthening was successful in 3 animals resulting in an increase in length of up to 696% from L1 to L5 compared to the growth rate of the controls (mean 4.3 cm vs. mean .56 cm, respectively). Lengthening of the spinal cord was not associated with brain stem migration or spinal cord injury. PMID- 8235841 TI - Intraoperative long-latency reflex activity in idiopathic scoliosis demonstrates abnormal central processing. A possible cause of idiopathic scoliosis. AB - Segmental reflex regulation in 37 patients with idiopathic scoliosis and 8 patients with nonidiopathic scoliosis was studied by recording evoked reflex muscle potentials from four muscle groups in each lower extremity during partial neuromuscular blockade. Effects on reflex activity mediated through descending systems arising in the brain stem were investigated by recording from proximal distal and flexor-extensor muscles. Ipsilateral and contralateral long-latency complex polysynaptic activity was present in all 37 patients with idiopathic scoliosis. This reflex activity was absent in eight nonidiopathic scoliosis patients. Long-latency reflex activity may represent segmental disinhibition. The presence of long-latency reflex activity in patients with idiopathic scoliosis and the absence of this activity in nonidiopathic scoliosis patients with curves of equal magnitude demonstrates that the curve per se is not responsible for the activity. This would imply that abnormal reflex processing may play a role in the development of the spinal deformity in patients with idiopathic scoliosis. PMID- 8235842 TI - Efficacy of spinal cord monitoring in neuromuscular scoliosis. AB - Somatosensory cortical evoked potentials (SCEPs) monitoring in neuromuscular scoliosis surgery was evaluated in a large consecutive series of spinal reconstructions to define its role in the detection and prevention of spinal cord injury; 141 procedures in 101 patients were evaluated. In 28% a reliable tracing could not be obtained. Two postoperative neurologic deficits were missed by SCEP monitoring. Diagnosis (Duchenne muscular dystrophy and polio) was the only significant factor found to correlate with a reliable tracing. The use of sublaminar wires was associated with a higher rate of change in tracing. Transient changes were common. SCEPs are unreliable and nonspecific in neuromuscular scoliosis surgery and are not efficatious in preventing or detecting spinal cord injury when used alone. Adjunctive techniques using epidural and MEPs must be studied in these patients. PMID- 8235843 TI - Demineralized bone matrix. Enhancement of spinal fusion. AB - A study was conducted to determine the ability of demineralized bone matrix gel to act as an osteoconductive/osteoinductive material to enhance canine spinal fusion. Seven dogs underwent posterior spinal fusion. Four-level fusions were performed with one of four procedures at each level: decortication alone, with gel added, with autograft, or with both gel and autograft. Dogs were killed at 6 weeks and early histologic response was studied. At untreated control sites, little bone formation was evident. Gel-filled sites showed abundant osteoid, with 60% of demineralized particles fused to or surrounded by new bone. Sites filled with autograft had more new bone, but there was more osteoid at gel-treated sites. Autograft augmented with gel showed the most vigorous response, with extensive bridging between demineralized particles, host bone, autograft, and new bone. Significantly less autograft was needed to induce a similar amount of new bone formation when gel was added. Use of the gel as an autograft extender may improve the chance for successful spinal fusion. PMID- 8235844 TI - A preliminary investigation of shape memory alloys in the surgical correction of scoliosis. AB - Nitinol, a shape memory alloy, is flexible at low temperatures but retains its original shape when heated. This offers interesting possibilities for scoliosis correction. Of the shape memory alloys, nitinol is the most promising medically because of biocompatibility and the ability to control transition temperature. In vivo: Six goats with experimental scoliosis were instrumented with 6-mm nitinol rods. The rods were transformed, and the scoliosis corrected, in the awakened goats by 450-kHz radio frequency induction heating. The curves averaged 41 degrees before instrumentation, 33 degrees after instrumentation, and 11 degrees after rod transformation. The animals tolerated the heating without discomfort, neurologic injury, or evidence of thermal injury to the tissues or the spinal cord. In vitro: Nitinol rods were tested under both constant deflection and constant loading conditions and plotted temperature versus either force or displacement. The 6-mm rod generated forces of 200 N. The 9-mm rod generated up to 500 N. We safely coupled shape memory alloy transformation to the spine and corrected an experimental spinal deformity in awake animals. The forces generated can be estimated by the rod's curvature and temperature. The use of shape memory alloys allows continuous neurologic monitoring during awake correction, true rotational correction by rod torsion, and the potential option of periodic correction to take advantage of spinal viscoelasticity and the potential of true rotational correction by rod torsion. PMID- 8235845 TI - Pathomechanical analysis of thoracolumbar burst fracture reduction. A calf spine model. AB - Burst fractures were created in the spines of twelve dairy calves and the mechanism of indirect canal clearance studied. The spines were reduced posturally and divided in to four subsets--control, and subsets in which either the posterior longitudinal ligament (PLL), the PLL and annulus, or posterior osteoligmentous structures were sectioned. The specimens were then instrumented and further reduction obtained through application of extension and distraction forces. Statistically significant reduction of the intracanal burst fragment occurred even if the PLL or PLL and annulus were incompetent but did not occur in that group in which the posterolateral complex had been sectioned. PMID- 8235846 TI - Treatment of severe spondylolisthesis by reduction and pedicular fixation. A 4-6 year follow-up study. AB - Ten consecutive patients with severe spondylolisthesis were treated with reduction and pedicular fixation (four Internal Fixator instrumentations, six Cotrel-Dubousset instrumentations). There were six Grade III spondylolisthesis and four spondyloptoses at the L5/S1 level. All patients had complete clinical and radiographic evaluation with an average follow-up of 56 months (range, 43-75 months). The percentage of slippage averaged 78.5% preoperatively and 39.6% postoperatively. The slip angle averaged 43 degrees preoperatively and 17 degrees postoperatively. Four patients with spondyloptosis were treated with combined posterolateral and interbody fusion and had solid fusion without loss of reduction. Five of six patients in whom reduction and stabilization was performed by a single posterolateral fusion demonstrated loss of reduction, nonunion, and implant failure. Four of these patients were reoperated. Ultimately all patients had resolution of pain, solid fusion, and no further slip progression. Reduction, pedicular fixation, and combined posterolateral and interbody fusion is a technically demanding procedure, which should be reserved for selected patients. Pedicular fixation systems may only allow permanent reduction and stabilization of high-grade spondylolisthesis in conjunction with a combined interbody and posterolateral fusion. PMID- 8235847 TI - The use of primary internal fixation in spondylolisthesis. AB - A prospective randomized study was designed to examine differences in the primary fusion rate between two surgically treated groups of patients with spondylolisthesis. Twenty-seven patients underwent L5 laminectomy (Gill procedure) and L5 nerve root decompression. Fourteen patients (group I) underwent in-situ posterolateral fusion, thirteen patients (group II) received internal stabilization with the Steffee plate and screw system. All patients were followed for a minimum of two years. Ten of fourteen (72%) patients treated with non instrumented fusion went on to union. Ten of thirteen (78%) patients treated with internal fixation achieved union. No statistically significant increase in the primary fusion rate occurred with addition of internal fixation compared to non instrumented posterolateral grafting alone. PMID- 8235848 TI - Pedicle screw pullout strength. Correlation with insertional torque. AB - This study was designed to correlate several parameters regarding pedicle screw bone/metal interface strength. The insertional torque measured during tapping and placement of pedicle screws was correlated with the bone mineral density of the vertebral body, the dimensions of the pedicle, the method of preparation of the pedicle, and the amount of load and number of cycles to failure of the bone/metal interface. Thirty human cadaveric lumbar vertebrae were instrumented with 6.5-mm pedicle screws. The maximum torque achieved during insertion was digitally recorded. A cyclic pedicle screw pullout test was performed. A linear correlation existed between both the insertional torque when tapping or when inserting a screw and the number of cycles to ultimate pedicle screw pullout. An inverse linear relationship was found between the pedicle width and cycles to failure. There was no linear correlation found when comparing the number of cycles to failure to bone mineral density. These findings suggest that insertional torque is a good predictor of bone-metal interface failure. Bone mineral density of the vertebral body was less effective as a predictor of failure. Smaller pedicle width correlated with increased insertional torque and cycles to failure. This may explain why patients with osteoporosis on radiography may still obtain stable fixation with pedicle screws. Other factors, such as pedicle dimensions and shape, affect screw purchase as much as vertebral body bone density. Insertional torque less than 4.0 inch-pounds led to early pedicle screw pullout. This study forms the basis for the authors' clinical use of an instrumented torque screwdriver to measure insertional torque in the operating room. PMID- 8235849 TI - Static and cyclical biomechanical analysis of pedicle screw spinal constructs. AB - Biomechanical evaluation of twelve different spinal devices in vitro employing pedicle screws was performed using static (n = 5) and cyclical testing (n = 3) parameters. In general, the rank order of implant failures was similar between static and cyclical tests, performed at 600 N compressive load, 5 Hz, and 1 million cycles. The mean number of cycles to failure was higher for spinal instrumentation systems employing longitudinal rods than those using plates (ANOVA F = 16.94, P < .001). At 600 N, the compact Cotrel-Dubousset, TSRH, and ISOLA rod systems demonstrated mean cycles to failure ranging from 200,000 to 800,000 cycles. The remaining devices including Dyna-lok, Kirschner plate, and VSP devices had failures ranging from 50,000 to 210,000 cycles. Polyethylene cylinders representing vertebral bodies were used to eliminate the problems of biologic deterioration encountered with cadaveric spines (a full cyclical test to 1 million cycles required 56 hours), and thus to provide analysis of the weak portion of each spinal system. The failure of eleven of the twelve spinal systems occurred by fracture of a pedicle screw, most commonly at the junction of the upper screw thread and the collar (Kirschner, AO fixator, standard CD, ISOLA, and TSRH). However, in Dynalok and VSP systems, fracture of the threaded portion of the screw just posterior to the integral nuts was the most common screw fracture location. The compact CD system was the only spinal implant that consistently failed by fracture of the longitudinal spinal member (rod). The fatigue life of rod based systems was longer than plate based systems. These studies confirm the importance of anterior column load sharing (vertebral body, corpectomy bone graft) as the mean bending strength demonstrated by these implant systems was not inordinately high using this "worst case scenario" model. PMID- 8235850 TI - An anatomic, radiographic, and biomechanical assessment of extrapedicular screw fixation in the thoracic spine. AB - This study performed a thorough review of the pertinent thoracic spine anatomy to determine the appropriate landmarks for extrapedicular screw placement and indentified the anatomic structures at risk; assessed the safety and accuracy of the newly proposed screw insertion technique; determined the stability and strength of this new screw position through biomechanical testing; and analyzed the stability of this extrapedicular screw location with respect to bone mineral density of the vertebra. PMID- 8235851 TI - Longitudinal element size effect on load sharing, internal loads, and fatigue life of tri-level spinal implant constructs. AB - The effects of implant stiffness on load sharing and stress shielding, of vertebral column load sharing on implant fatigue life, and of instrumenting two versus one level adjacent to a comminuted segment on implant internal loads were studied. Finite element models of six screw constructs with 4.76 mm rod; 6.35 mm rod, and VSP plate tri-level instrumentation of two motion segments (healthy vertebra case and comminuted) and an adjacent healthy motion segment with dimensions representative of the human lumbar spine were used. Also a simplified model was developed to predict the percent of axial load passing through the column, which is a function of ki/kv the ratio of implant axial stiffness to instrumented vertebral column axial stiffness. For constructs with dimensions typical of the human lumbar spine, 77 to 80% of the axial load was predicted to pass through one or two healthy motion segments when instrumented with either 6.35 mm rod or VSP plates, compared to 90% when instrumented with 4.76 mm rods. When instrumenting smaller motion segments (in dogs) for comparison, 60% of the axial load was predicted to pass through the column for 4.76 mm rod and 33% for 6.35 mm rod constructs due to increased implant stiffness ki as a result of decreased AP and longitudinal construct dimensions, and lower canine motion segment stiffness kv.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235852 TI - 17-year prospective study of surgical management of spinal tuberculosis in children. Hong Kong operation compared with debridement surgery for short- and long-term outcome of deformity. AB - A comparative analysis of the short and long-term results of two surgical procedures, radical excision or debridement, for the management of spinal tuberculosis in children is reported. Eighty children (47 treated with radical surgery and 33 with debridement) were prospectively studied and followed to maturity (mean follow-up, 17 years postoperatively). The kyphus and deformity angles were measured from lateral spinal radiographs using an electronic digitizer. The results can be summarized as follows: 1) Long-term clinical outcome of the two surgical procedures were equally good for recovery of neurologic deficit and relief of pain. There was no incidence of reactivation and/or recurrence of tuberculous lesion in either group; 2) The mean changes in kyphus and deformity angles at the 6-month postoperative evaluation compared to their preoperative value were significantly different for the two surgical groups. There was an overall correction in these angles after radical surgery, whereas there was a deterioration after debridement surgery; 3) There were no significant differences in the changes in kyphus or deformity angles at final follow-up from their 6-month postoperative measurements between the two surgical groups; 4) The majority of children (56%) showed an improvement in deformity angle of 5 degrees or more after radical surgery at the 6-month postoperative evaluation, whereas 69% of children showed deterioration after debridement surgery; 5) At final follow-up in lumbar tuberculosis, 60% of patients in the debridement group had 10 degrees or more kyphus angle, whereas only one patient in the radical group had a kyphotic lumbar spine due to graft failure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235853 TI - The pain drawing and Waddell's nonorganic physical signs in chronic low-back pain. AB - Pain drawings were obtained from a group of 651 patients who had chronic low-back pain. Pain drawings were rated in four grades according to the degree of organic and nonorganic pain. Experienced and inexperienced evaluators were used. The reliability was excellent with an interevaluator reliability between 73% and 78%. A correlation between pain drawings and Waddell's nonorganic physical signs demonstrated that a large proportion of patients with high Waddell scores had nonorganic pain drawings. No significant differences were noted in the distribution of Waddell scores and pain drawings based on patient gender or payment status (i.e., medicolegal or workers' compensation). Pain drawings afford an important adjunct in the physician's assessment of chronic low-back pain. PMID- 8235854 TI - Sensitivity of Sickness Impact Profile items to measure change over time in a low back pain patient group. AB - The purpose of this study was to identify Sickness Impact Profile (SIP) items that are most sensitive to change in patients with low-back pain. Seventy-six patients with low-back pain were administered the SIP at their initial visit and after discharge from physiotherapy treatment. A formal item reduction was performed to identify the most sensitive items. An item was considered sensitive if it showed change in 20% of the patients and had an item-corrected total SIP score correlation greater than 0.30. Twenty items were identified. Seven of the 20 items identified in this study appear on the Roland-Morris disability questionnaire. Also, only 50% of the items identified are from the physical subscale of the SIP. PMID- 8235855 TI - Nonsurgical hospitalization for low-back pain. Is it necessary? AB - In 1988, the Diagnosis-Related Group for "Medical Back Problems" was the seventh leading reason for all U.S. hospitalizations. The authors sought to describe the content of these hospitalizations and consider the potential for shifting nonsurgical care to the outpatient setting. Three complementary data sources (the 1988 National Hospital Discharge Survey, a statewide Washington hospital discharge registry, and medical records) were used to examine the diagnoses, tests, treatments, resource use, and subsequent care associated with these hospitalizations. Nationally, nonspecific back pain and herniated discs were the most common diagnoses. Nearly half the hospitalizations were for diagnostic tests (especially myelography) and the other half for pain control. In Washington state, 43% of patients were admitted by family physicians or internists, and 40% by orthopedic or neurologic surgeons. Twenty percent of patients underwent subsequent back surgery within 1 year (most within 3 months), suggesting that many hospitalizations were "presurgical." Most of the tests and treatments identified are known to be safe in the outpatient setting. Focused medical record review indicated frequent psychosocial problems or complicating factors, including obesity, substance abuse, prior back surgery, psychologic diagnoses, or lack of a caregiver at home. The findings support other evidence that many hospitalizations for "medical back problems" are unnecessary, but also suggest a need for improved outpatient and home-based alternatives to hospitalization. PMID- 8235856 TI - The development of guideline factors for the evaluation of disability in neck and back injuries. Division of Industrial Accidents, State of California. AB - This is the second article documenting the development of guideline factors for the evaluation of spine injuries by a committee supported by the Division of Industrial Accidents (DIA) in the State of California. The committee was asked to develop guidelines that incorporated cervical and thoracic spine injuries into the original guidelines developed for lumbar spine injuries (Clark et al, Spine 1988; 13:332). Multiple Independent Medical Examiners (IMEs) were requested to rate disability on 42 case reports submitted to the DIA. A tremendous variation existed in the disability rating recommended by physicians given the same set of facts. One hundred eighty-five IMEs then were asked to evaluate a series of potential factors influencing disability. Each factor was listed according to its perceived importance in determining disability. These factors then were subjected to an intensive search of the medical and scientific literature to determine their validity. A final list of 28 factors for the cervical and thoracic spine were developed and combined with the previously published factors for the lumbar spine. This led to the development of 37 factors or guidelines for the evaluation of spine disability. Testing of this schedule was performed by reviewing 159 submitted disability consultation reports. This review showed that there were no factors in the reports that were not included in the schedule. An average of 3.1 subjective factors and 2.3 objective factors were noted in the reports, with a very poor relationship between the number of factors and the level of disability given by the assessing physician. It is thought that the incorporation of the proposed guideline factors into the current disability rating system would allow for a more reproducible evaluation of disability and allow Workers' Compensation judges and administrators to make disability decisions based on documentable and reliable facts. PMID- 8235857 TI - Dorsal root ganglia in the lumbosacral region observed from the axial views of MRI. AB - A total number of 442 dorsal root ganglia (DRG) in the lumbosacral region of 104 patients were depicted from the axial views of MRI and their localities were classified as intraspinal, intraforaminal, or extraforaminal types. The DRG of the more distal nerve root were located more medially. Asymmetrical locations were observed in 25 pairs of DRG among 221, of which the symptomatic side tended to locate either medially or proximally compared with those on the asymptomatic side. PMID- 8235858 TI - Increase of serum growth hormone-binding protein in patients with ossification of the posterior longitudinal ligament of the spine. AB - Serum growth hormone-binding protein (GHBP) was measured in 26 patients with ossification of the posterior longitudinal ligament of the spine (OPLL) and 19 age-matched controls. Serum GHBP level was significantly higher in the OPLL group than in the controls, whereas there was no statistical difference between the two groups in serum level of growth hormone, insulin-like growth factor (IGF)-1, and IGF-2. These results, taken together with the hypothesis that serum GHBP reflects the number of GH receptors in tissue, suggest that GH receptors are increased in patients with OPLL. PMID- 8235859 TI - Neutral proteinases in human intervertebral disc. Role in degeneration and probable origin. AB - Neutral proteinases were partially purified from human intervertebral disc by extraction with guanidine hydrochloride followed by Sephadex G-75 column chromatography. They showed gelatinolytic and elastolytic activities at neutral pH. The apparent molecular weights of these enzymes were 70 KD and 25 KD, the former being the complexed form of the latter. The sensitivity to various synthetic inhibitors indicated these enzymes to be serine elastases. Furthermore, they were considered to be leukocyte elastases because anti-leukocyte elastase antibody inhibited the proteinase activity of these enzymes. Proteinase activity was not detected in normal discs, except for some slight activity in the end plate. In contrast, the degenerated discs showed high activity, especially in the nucleus pulposus and end-plate. Similar high levels of proteinase activity were found in the vertebral body adjacent to normal as well as degenerated discs. These findings suggest that leukocyte elastase, which is normally present in the vertebral body, flows into and degrades the matrix of the intervertebral disc under pathologic conditions. PMID- 8235860 TI - Effects of laminectomy and retained extradural foreign body on cauda equina adhesion. AB - The histologic effects of posterior lumbar surgery and retained extradural foreign bodies on the cauda equina were investigated in rats over time. The following four groups of rats were provided: Group 1, sham operation (laminar exposure alone); Group 2, laminectomy alone; Group 3, laminectomy with retained extradural silk thread; Group 4, laminectomy with extradural kaolin; and the control. Histological study was performed on the transverse sections of laminectomized (L5) and nonlaminectomized (L4) areas. In Group 1, adhesive changes involving the cauda equina, consisting of dilatation of nutrient vessels, and eosinophilic exudative changes between the adhering cauda rootlets were seen the day after surgery, but the fibrinous adhesion was resolved spontaneously in all rats after 1 week without permanent sequelae. In Groups 2 and 3, postoperative adhesion of the cauda equina roots was conspicuous, but it resolved by 6 weeks. In Group 4, obvious adhesion persisted through the 12th week after surgery. Destruction of the myelin sheath and axonal loss of the cauda equina occurred in the first week after surgery in all the experimental groups except in the sham group. The severity and extent of the neural degeneration were mostly parallel to the severity of inflammatory changes in the epidural areas. At the nonlaminectomized area (L4) in each group, the arachnoid and cauda equina tended to congregate the day after surgery, but these changes were all transitory. The severity and persistency of the arachnoiditis and neural degeneration directly corresponded to the magnitude of the inflammation and wound healing processes. This seems to imply that careful surgical intervention with no use of foreign bodies is necessary to reduce postoperative adhesive arachnoiditis. PMID- 8235861 TI - Study on dermatomes by means of selective lumbar spinal nerve block. AB - Regions of sensory impairment of 71 patients in 86 lumbar spinal nerve blocks (L4 19 cases, L5 41 cases, S1 26 cases) were determined by the writing brush method. Distinctive regions included the medial side of the lower leg, the side of the first dorsal digit, and the side of the fifth digit respectively of the L4 (88%), L5 (82%) and S1 (83%) spinal nerves without 100% matching because of displacement of one spinal segment by the furcal nerve. A band-like impaired zone from the proximal to the periphery appeared in 42%, 44%, and 92%, respectively, of the L4, L5, and S1 blocks. The marked difference between L4, L5, and S1 may be caused by the diversity of the dorsal rami of the spinal nerves and double control by the lateral cutaneous nerve of the thigh. PMID- 8235862 TI - Effect of electrical stimulation on lumbar paraspinal muscles. AB - Forty healthy, sedentary, premenopausal women were entered into a randomized, single-blind, controlled study to determine the effects of capacitively coupled electrical stimulation on the strength of the lumbar paraspinal muscles and the bone mineral in the lumbar spine. All were between 35 and 45 years of age and had normal physiologic estrogen. The study group received electrical stimulation over the lower lumbar paraspinal muscles for 30 minutes twice a day. Isometric strength of the lumbar paraspinal muscles was assessed with a strain-gauge dynamometer at entry and again after 3, 6, 9, and 12 months. Bone mineral was measured in the lumbar spine by dual-energy x-ray absorptiometry at entry and at 6 and 12 months. After 3 months, the median increase in isometric strength was 8.1% in the study group and 1.6% in the control group (P < 0.03). This initial difference was maintained during the remainder of the study. No further changes were seen between the two groups at 6, 9, and 12 months. It was concluded that capacitively coupled electrical muscle stimulation can, throughout a 1-year period, improve and maintain isometric strength of the lumbar paraspinal muscles independent of exercise, but it has no measurable effect on bone mass in the lumbar spine. PMID- 8235863 TI - The efficacy of femoral nerve intraoperative somatosensory evoked potentials during surgical treatment of thoracolumbar fractures. AB - Somatosensory evoked potential (SEP) monitoring of thoracolumbar procedures typically includes posterior tibial and peroneal nerve recordings. The addition of femoral nerve SEP monitoring, however, should better predict the evolution of postoperative neurologic deficits affecting the midlumbar roots. To assess the value of intraoperative femoral SEPs, 26 cases of traumatic injury to the thoracolumbar spine between T12 and L4 were prospectively assessed. Twenty-four had clear femoral nerve responses bilaterally recorded from an epidural electrode. Five had significant intraoperative SEP changes: one had isolated femoral nerve changes, two had only peroneal or tibial nerve changes, and two had concomitant changes in both femoral and peroneal or tibial nerves. Loss of the femoral nerve response in one patient was correlated with marked postoperative knee extensor weakness, in spite of immediate action taken by the surgeon. The authors conclude that femoral nerve SEPs provide an effective tool to monitor the midlumbar roots intraoperatively. PMID- 8235864 TI - Effects of electrical stimulation of motor and cutaneous nerves on spinal cord blood flow. AB - The effects of electrical stimulation of the motor and cutaneous branch of the radial nerve on spinal cord blood flow (SCBF) in the cervical region and the correlation between the changes in SCBF and the amplitudes of spinal cord evoked potentials were investigated in cats anesthetized with alpha-chloralose. When measured by the hydrogen clearance technique, with stimuli of the cutaneous branch, the SCBF significantly (P < 0.01) increased from 37 +/- 8 (mean +/- SD) to 59 +/- 27 ml.100 g-1.min-1 at 10 Hz and from 39 +/- 6 to 54 +/- 9 ml.100 g 1.min-1 at 30 Hz, but stimulation of the motor branch had no effect. When measured by laser Doppler flowmetry, stimulation of the motor branch caused a small, transient, and significant increase in SCBF ending within 2 minutes. Stimulation of the cutaneous branch produced a large, prolonged, and significant increase in SCBF. The increase in SCBF produced by stimulation of the cutaneous nerve was significantly greater than that by stimulation of the motor branch. The SCBF was correlated linearly with the amplitude of the spinal cord evoked potential (r = 0.504; P < 0.01). The increase in SCBF caused by radial nerve stimulation may arise from changes in neuronal metabolism via peripheral nerve activation. PMID- 8235865 TI - Myofascial trigger points show spontaneous needle EMG activity. AB - Monopolar needle electromyogram (EMG) was recorded simultaneously from trapezius myofascial trigger points (TrPs) and adjacent nontender fibers (non-TrPs) of the same muscle in normal subjects and in two patient groups, tension headache and fibromyalgia. Sustained spontaneous EMG activity was found in the 1-2 mm nidus of all TrPs, and was absent in non-TrPs. Mean EMG amplitude in the patient groups was significantly greater than in normals. The authors hypothesize that TrPs are caused by sympathetically activated intrafusal contractions. PMID- 8235866 TI - The symptomatic lumbar disc in patients with low-back pain. Magnetic resonance imaging appearances in both a symptomatic and control population. AB - Magnetic resonance imaging of abnormal lumbar discs in 115 symptomatic patients (184 discs) and 63 symptom-free controls (78 discs) have been compared. All symptomatic patients were proven to have discogenic pain at discography. Discs were classified into patterns 1-6 reflecting the progressive changes of nuclear degeneration as previously described. In the symptomatic patient, the more degenerate the disc the more likely it was responsible for pain (P < 0.001), but there was no statistically significant difference in the distribution of abnormal signal patterns between the two groups studied. A small number of patients in both populations demonstrated high signal from disc protrusions but this also could not be used as a predictor of pain. No abnormal lumbar disc signal pattern could be identified that specifically indicated whether a disc would be painful. Magnetic resonance imaging is accurate in determining nuclear anatomy, but until more suitable techniques become available, discography is still the only method for symptomatic assessment in low-back pain. PMID- 8235867 TI - The role of T2-weighted gradient echo in MRI demonstration of spinal multiple myeloma. AB - Fifty patients with proven multiple myeloma (MM) underwent magnetic resonance (MR) examination of entire spine in sagittal view using T1-weighted image (T1), T2-weighted image (T2), and T2-weighted gradient echo (GE). In 18 patients, the myelomatous foci were hyperintensive in GE and T2 and hypointensive in T1. They corresponded with osteolytic lesions in computed tomography (CT) scan. In another 16 patients with MM, the hyperintensive vertebral foci demonstrated by GE, corresponded with unhomogenic pattern of the vertebral bone marrow in T1 and T2, and insignificant computed tomography (CT) scan. Needle biopsy confirmed histologically the diagnosis MM of involved vertebra in all of them. The vertebral bone marrow had lower signal intensity in GE, which ensured a good contrast between the myelomata and the uninvolved bone marrow. Practically GE enabled the radiologic diagnosis of the spine in 16 patients. It also can prevent a diagnostic pitfall when a fatty focus is suspicious for myeloma in T2 and its demonstration in T1 is poor. Fatty foci were demonstrated in a control group, which included 20 elderly patients who had no history of malignancy. The fatty foci had lower signal intensity in GE and different from the hyperintensive myelomata. Gadolinium diethylene triamine pentaocetic acid (Gd-DTPA) which was administered intravenously in three patients with spinal MM offered no advantage and obscured the lesions. PMID- 8235868 TI - Intraoperative use of the mobile gamma camera in localizing and excising osteoid osteomas of the spine. AB - Three patients (ages 12, 13, and 15 years) had histologically verified osteoid osteomas of the spine (specifically in the facet processes of L3, L2, and C5) which were localized preoperatively with nuclear scintigraphy utilizing technetium 99m diphosphonate. Intraoperative nuclear scintigraphy utilizing the mobile gamma camera was essential in both localizing the tumor during surgical excision and ensuring adequacy of excision. The authors strongly recommend intraoperative scintigraphy, and en bloc resection rather than shelling out and curettage. All three patients required facet process excision for successful tumor removal, and therefore required local fusion with internal fixation. PMID- 8235869 TI - The effects of resistance level on muscle coordination patterns and movement profile during trunk extension. AB - The objective of this research was to investigate the effects of varying resistance level on patterns of muscle activity and trunk motion during the performance of dynamic trunk extension. Twenty-five female subjects were tested at four resistance levels: nominal resistance, and 30%, 50%, and 70% of maximum isometric extension strength. Peak truncal velocity, acceleration, and deceleration decreased with increased resistance, although no difference existed between the nominal and 30% maximum voluntary contraction (MVC) conditions. Coactivation of abdominal muscles was present at all resistance levels, although it was greater for the nominal resistance condition than the other conditions. The timing of EMG activity of the extensor and abdominal muscles was significantly affected by resistance. The amplitude and time course of peak muscle activity are discussed in terms of the implications for spinal loading. PMID- 8235870 TI - Efficacy of flexion and extension treatments incorporating braces for low-back pain patients with retrodisplacement, spondylolisthesis, or normal sagittal translation. AB - Radiographic instability seemingly enjoys the status of a well-defined clinical syndrome. The concept is widely used, and specific treatments, usually spinal fusion, are routinely performed based on the diagnosis. The minimum standards necessary to establish radiographic instability as a legitimate clinical syndrome have not been established, however. The primary purpose of this study was to determine if treatment involving bracing, exercise, and education controlling either flexion or extension postures, would result in a distinctive pattern of favorable or unfavorable results, depending on the type of radiographic instability (retrodisplacement or spondylolisthesis). Fifty-six patients meeting strict study inclusion and radiographic evaluation criteria were assigned signed to a bracing treatment (flexion, extension, placebo-control) according to a randomization scheme, designed to ensure equal representation of translation categories (retro, normal, spondy) across treatment groups, and assessed at admission and 1-month follow-up. The sample was relatively evenly divided between men (46%) and women (54%), and by age. Translation classification was related to both gender and age, with men more likely classified as retro and women more likely spondy and patients in their 20s having lower incidence of spondy and higher incidence of normal translation. Translation classification was not related to selected indices of low-back pain history. Brace treatments were not shown to reduce patient range of motion or lessen trunk strength. A significant treatment by time interaction for the modified pain interference (VAS) scale indicated improvement for patients in extension compared with patients in flexion and control-placebo treatments. In conjunction with no significant three-way interaction between treatment, translation classification, and time, it was hypothesized that radiographic instability might more appropriately be considered a corroborative sign of advanced discogenic problems. Improvement in extension treatment, regardless of the type of radiographic abnormality, suggests that the treating clinician might consider extension treatment for chronic low-back pain patients. Causes and implications for the failure of this study to provide support for considering radiographic instability as a clinical syndrome are considered and future directions for this area of research suggested. PMID- 8235871 TI - Pedicle morphology of the lower thoracic and lumbar spine in a Chinese population. AB - Knowledge of pedicle diameter and surface landmarks is crucial for safe placement of screws. Little attention has been paid to variations of entrance points for pedicle screws, differentiation of male and female pedicle sizes and pedicle size differences in nonwhite populations. Forty thoracolumbar spinal columns from T9 to L5 were measured using vernier calipers. Cephalad-caudad and medial-lateral diameter of the pedicle, length of the pedicle from posterior cortex to anterior cortex at the midline and parallel to midline was measured. Relation of the centre of the pedicle to the transverse process (TP) and to the superior facet joint was noted. Twenty-five male and 15 female specimens were measured. Average pedicle width in the female was 5.2 mm at T9 (SD 0.9) to 13 mm at L5 (SD 2.7) and in the male 6.0 mm at T9 (SD 1.1) to 12.8 mm at L5 (SD 2.7). Cephalad caudad diameter was 12.5 mm (SD 1.2) at T9 to 20.5 mm (SD 3.6) at L5 in the male and in the female 12.2 mm (SD 1.3) at T9 to 18.7 mm (SD 3.9) at L5. All specimens had starting points cephalad to the midpoint of the TP at T9. At L5, 37 of 40 specimens had starting points at the midpoint of the TP. Starting points were parallel to the middle or lateral third of the superior facet joint at T9. At L5 starting points were at least one third of the facet joint lateral to the lateral border of the facet. Female pedicle width was smaller than male at T9 (P = 0.03) and T12 (P = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235872 TI - Spinal pedicle fixation. Confirmation of an image-based technique for screw placement. AB - Although many advances in the technique of pedicle screw insertion have been made, there still exist unacceptable rates of perforations through the pedicle cortex. Successful placement of a pedicle screw requires accurate identification of the entry point, correct transverse and sagittal plane angulation, safe preparation of a pilot hole, and appropriate depth of insertion. The authors propose that a technique of pedicle screw insertion using posteroanterior image intensification angled in the axis of the pedicle to define the entry point and guide insertion would improve the accuracy of this procedure. This study tests the accuracy of pedicle screw placement using this technique. Ninety pedicle screws were placed in human cadaveric lumbar spines. All specimens were then dissected and split longitudinally to assess accuracy of pedicle insertion by both visual and palpatory means. Five of 90 (5.5%) pedicle screws were found to have perforated the pedicle. Three of these were at L1, two of which were due to the transverse pedicle diameter being smaller than the screw diameter. Therefore, a 3.4% incidence of pedicle perforation due to malpositioning and a 2.1% incidence of pedicle perforation secondary to pedicle/screw size discrepancy was determined. This technique may lead to significant improvements over recently reported rates of pedicle screw perforations; however, this in vitro condition eliminated many potentially complicating factors that might be encountered in the clinical setting. PMID- 8235873 TI - The effect of a stiff spinal implant and its loosening on bone mineral content in canines. AB - A canine spine surgery model was used to study bone mineral loss induced in vertebrae bypassed by stiff implants. Loosening of implants was measured, and a determination was made as to the effect of loosening on the degree of bone mineral loss. Sixteen dogs were implanted with a rigid screw-rod construct and randomized to one of four groups depending on the motion segments immobilized. No fusions were performed. Bone mineral was studied by DEXA at baseline, and postoperatively at 3, 6, and 9 months. Loosening measurements were made at the implant-implant interface and the implant-bone interface at 9 months postoperatively. Results show that bone mineral loss occurred uniformly at 3 months postoperatively in vertebrae spanned by the implants, with the average bone mineral density at 3 months representing 81% of baseline. When the implant remained secure, bone mineral loss persisted at 6 and 9 months but did not worsen. When the implants loosened, there was recovery of bone mineral to the baseline level by 9 months postoperative. PMID- 8235874 TI - Complications of the Wiltse Pedicle Screw Fixation System. AB - The purpose of the current study is to retrospectively review the incidence and types of complications occurring using a single pedicle screw fixation system for lumbar spine fusions. For a 3-year period beginning in July 1987, 470 patients underwent spinal fusion and instrumentation with the Wiltse Pedicle Screw Fixation System. Although multiple configurations of the Wiltse System were included, the predominant construct was that of segmental pedicle screw fixation. Complications occurred in a total of 29 patients, for an overall complication rate of 6%. Among the complications were 10 wound problems (2%) that included 2 seromas requiring drainage and 3 superficial wound infections. The remaining five wound complications were deep wound infections requiring irrigation and debridement. The overall deep wound infection rate was 1%. Neurologic complications occurred in two patients, one with temporary foot drop and one with persistent L5 nerve root irritation confirmed by electromyogram (EMG). Both of these patients underwent anatomic workup, which failed to reveal any evidence of impingement by either screw or other hard tissue. Two intraoperative complications occurred, both involving fracture of the L5 pedicle. No adverse sequela resulted. There were 15 hardware complications, for an overall rate of 3%. This included four patients with broken screws (a total of eight screws). There were three failures at the screw-bone interface, as well as four patients with uncoupling of the screw rod linkage. In addition, there were four broken rods encountered. A total of 122 Wiltse devices were removed, for an overall removal rate of 25%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235875 TI - Thoracic disc herniation. Re-evaluation of the posterior approach using a modified costotransversectomy. AB - A consecutive series of 23 thoracic disc herniations in 21 patients treated between 1980 and 1988 were reviewed. All patients were decompressed through a posterolateral approach (costotransversectomy or transpedicular). Pain and weakness were the most common presenting symptoms. Twenty-one thoracic disc herniations in 19 patients were available for long-term follow-up, averaging 58.1 months. Sixteen patients had an excellent or good result. Three patients had a fair result. There were no poor results. All six patients with significant preoperative lower extremity weakness improved. Pain was relieved in 16 patients and reduced in three. There were no significant neurologic complications associated with the procedure. Posterolateral decompression for thoracic disc herniation remains a viable alternative without the inherent risk and morbidity of the transthoracic approach. PMID- 8235876 TI - Burst fractures of the fifth lumbar vertebra. AB - Eleven patients with burst fractures of the fifth lumbar vertebra were reviewed. The results of nonoperative treatment were compared with that of immediate surgery and stabilization with pedicle screw fixation. Five patients were treated nonoperatively and six patients underwent pedicle screw instrumentation and spinal fusion. Five patients had neurologic injury associated with their L5 burst fracture. Nonoperative treatment yielded excellent results in young patients with minimal canal compromise. Neurologic deficits responded more predictably to surgical decompression than to conservative treatment and internal fixation with pedicle screws restores spinal stability and allows early mobilization. PMID- 8235877 TI - Observations of resorption of the posterior lateral bone graft in combined anterior and posterior lumbar fusion. AB - The past 10 years have seen the development of a surgical reconstructive procedure for the treatment of patients disabled with severe postlaminectomy syndrome. The technique involves anterior lumbar fusion, replacing the excised disc with bone graft, and under the same anesthetic, applying a posterolateral bone graft along with segmental instrumentation. The purpose of this report was to compare cases with fused and nonfused posterolateral grafts in cases where the anterior fusion was successful. Seventy-four interbody segments in 30 patients were fused while 47% (14 of 30) of the posterolateral bone grafts underwent resorption. Both allograft and autograft were used and this did not appear to influence the resorption seen at the 4-year follow-up period. The most likely factor to explain resorption of the posterolateral graft is deprivation of the proper mechanical forces in the lateral gutter by the simultaneous use of anterior lumbar interbody grafts. PMID- 8235878 TI - Evaluation of the risk of instrumentation as a foreign body in spinal tuberculosis. Clinical and biologic study. AB - The risk of persistence and recurrence of infection in posterior spinal instrumentation surgery for spinal tuberculosis was studied clinically and microbiologically. Eleven patients with thoracic, thoracolumbar, and lumbar spinal tuberculosis treated by debridement, anterior fusion, and combined posterior instrumentation surgery were analyzed. Seven patients had tuberculosis in both anterior and posterior spinal elements. There were no cases of persistence or recurrence of infection after surgery, and instrumentation provided immediate stability and protected against development of kyphotic deformity. The adherence properties of Mycobacterium tuberculosis to stainless steel (SUS 316) was evaluated experimentally. The results showed that posterior instrumentation surgery was not a hazard to spinal tuberculosis infection when combined with radical debridement and intensive anti-tuberculosis chemotherapy. PMID- 8235879 TI - The pullout strength of titanium alloy MRI-compatible and stainless steel MRI incompatible Gardner-Wells tongs. AB - The stainless steel Gardner-Wells tongs and pins previously employed to reduce cervical spine fractures are increasingly being replaced by magnetic resonance imaging (MRI)-compatible graphite tongs and titanium pins. Concern regarding the relative pullout strengths of these two systems, however, prompted the performance of a cadaver study in which tongs and pins were placed in 10 cadaver skulls and tested to failure. The MRI-compatible tongs failed due to plastic deformation of the titanium pin tips resulting in tong slippage and loss of fixation, whereas the stainless steel tongs and pins failed when the skull fractured. These results indicated that MRI-compatible systems must be used with extreme caution, particularly when traction exceeds 50 pounds. PMID- 8235880 TI - Traumatic atlanto-occipital dislocation with survival. Serial computerized tomography as an aid to diagnosis and reduction: a report of three cases. PMID- 8235881 TI - One-stage "front" and "back" correction for rigid cervical kyphosis. A safer technique of correction for a rare case of adult-onset Still's disease. AB - This report describes a new safe technique for surgical correction of rigid cervical kyphosis, in a very rare case of adult-onset Still's disease. It differs from all the previously mentioned reports of similar corrective osteotomies, the essential difference being a combined anterior (front) and posterior (back) approach performed at one stage (under a single anesthesia). The "front" is a release procedure of an open-wedge variety, and the "back" is a closed-wedge type of osteotomy. Such a double procedure ensures a slow and controlled correction at the preoperatively determined level, rather than a snapshot, rapid, and often a violent correction, which may occur at an undesired level. This osteotomy is performed under general anesthesia after an endotracheal intubation by a special "guided" maneuver. This new technique, the authors believe, is only a more comfortable option for the surgeon, but also a safer alternative for the patient. PMID- 8235882 TI - Oligosegmental correction of post-traumatic thoracolumbar angular kyphosis. AB - Seventeen patients with rigid throacolumbar angular kyphosis due to neglected fractures or dislocations were treated by a standardized single-stage monosegmental or bisegmental anterior discectomy and posterior closing extension wedge osteotomy. The two- or three-level reduction-fixation (RF) instrumentation was used posteriorly for correction and fixation. Choosing angled pedicle screws according to preoperative measurements, the method can always correct the kyphosis to the approximate sagittal curvature that is planned to create preoperatively. The average precorrection of thoracolumbar kyphosis was 39 degrees and was restored to 1.2 degrees for an average correction of 37.8 degrees (range, 22 degrees-56 degrees) with subsequent average loss of 1.1 degrees at final follow-up. Before operation, the complaints were slow progression of kyphotic deformities, fatigue, and pain. All these problems were solved by this procedure. Complications were minimal and mild. No neurologic complications occurred. Follow-up averaged 2.8 years. This method can correct rigid post traumatic thoracolumbar angular kyphoses to normal geometric relationships as planned preoperatively without much negative effect in lumbar motion and any sacrifice of safety. PMID- 8235883 TI - The sternal-rib complex. A possible fourth column in thoracic spine fractures. AB - Two cases are described in which displaced sternal fractures were associated with thoracic spine fractures of minimal, initial displacement. Nonoperative, expectant treatment led to significant kyphotic deformity. It is postulated that the sternum and ribs represent a fourth column of structural thoracic spine support. An overriding, displaced sternal fracture is a marker for a severe flexion-distraction unstable thoracic spine injury with a propensity for deformity. PMID- 8235884 TI - Hematometra as a cause of lumbar radiculopathy. A case report. AB - Low-back pain combined with a positive straight leg raising test (Lasegue's sign) is uncommon in children and adolescents. Reproductive tract (Mullerian) anomalies causing an accumulation of menstrual blood in the vagina, uterus, and fallopian tubes represent an unusual extraspinal cause of low-back pain. The accumulation of blood in these cavities is termed hematocolpos, hematometra, and hematosalpinx. Amenorrhea with cyclic lower abdominal pain and a presenting pelvic mass are the usual presenting features. In the current report, we describe a case of an adolescent with low-back pain, positive straight leg raising test, and signs of L5 radiculopathy that were attributable to hematometra secondary to congenital partial absence of a vagina. This appears to be the first report of this type of presentation. PMID- 8235885 TI - Acrodysostosis associated with spinal canal stenosis. AB - An adult case of acrodysostosis with striking lumbar spinal canal stenosis is reported. She complained of numbness on the right arm, intermittent claudication with numbness on both legs and pain and weakness on the left leg. Although the reduced lumbar interpedicular distance has been reported, no adult case with neurologic symptoms has been reported. PMID- 8235886 TI - Spinal osteomyelitis due to Mycobacterium avium-intracellulare in an elderly man with steroid-induced osteoporosis. AB - Disseminated mycobacterial infections are becoming widely appreciated among immunosuppressed and debilitated persons. Specific microbiologic diagnosis may take several weeks, requiring empiric therapy in the interim. A case of an elderly man undergoing steroid treatment who developed spinal osteomyelitis initially thought to be caused by Mycobacterium tuberculosis is described. He was eventually shown to have infection due to M. avium-intracellulare, which requires a different therapeutic approach. The importance of considering atypical mycobacteria in systemic infections and the value of aggressive antimicrobial therapy in conjunction with a comprehensive rehabilitation program during hospitalization are emphasized in order to provide optimal management and prevent further disability. PMID- 8235887 TI - [Lyme disease in Mexico City]. PMID- 8235888 TI - [The evolution of the health services of the Secretaria de Salud]. PMID- 8235889 TI - [An epidemic of primary bacteremia due to an endemic strain of Serratia marcescens in an intensive care unit]. AB - An outbreak of Serratia marcescens bacteremia detected in the intensive care unit (ICU) of a tertiary care center on the last days of October, 1985, is described. The rate of primary S. marcescens nosocomial bacteremia during the pre-epidemic period (January-September 1985) was 6.25 per cent; and for the post-epidemic period compared with the epidemic were significantly different (p < 0.0001). The outbreak strains belonged to the biotype A8b, which has been endemic in our hospital. The responsible organism exhibited an unusual antimicrobial resistance pattern associated to the presence of a specific plasmid (greater than 50 kilobases), which showed similar fragments after restriction endonuclease digestion. No specific risk factors were identified in the case-control study. The outbreak was probably related to a greater influx of infected patients, resulting in less careful infection control measures, due to the emergency situation which suffered the hospital after the earthquakes in 1985. The unusual high rate of blood isolation of S. marcescens at the ICU was the first sign of the outbreak. The prompt reinforcement of infection control policies facilitated its resolution. PMID- 8235890 TI - [Accidents and injuries in 4 general hospitals of the Distrito Federal]. AB - We report the injuries for "E" code in 10191 subjects attending four general hospitals of Mexico City. Seventy-nine per cent of them were males and 29.1 per cent were females. THe highest utilization rate of the care unit was during weekends. Only 14 per cent of the study population were admitted to the hospital. People classified as drunk were more likely to have more severe injuries and to be admitted to the hospital and female drinkers had the highest risk of hospitalization (RR = 3.03; IC = 1.85-5.04). Only in 49 per cent of the cases was the external cause of injury coded The main causes were gunshots and stabbing with knife or cutting weapon. In this study we found a greater proportion of fractures in women than in men, pointing toward the possible presence of osteoporosis in the female population. PMID- 8235891 TI - [Outbreaks of food poisonings of microbial and parasitic origins]. AB - In order to know the agents and foods related more frequently with food-borne disease outbreaks, we reviewed all the outbreaks studied between 1980 and 1989 by the National Laboratory of Public Health. A total of 79 outbreaks of food-borne diseases of microbial origin were reviewed. The causative agent was identified in 50 (73%) outbreaks. Twenty-four per cent of the outbreaks occurred in parties, 10.3 per cent in school or nurseries, 8.6 per cent in restaurants and 8.6 per cent in hospitals. Staphylococcus aureus was the most common agent, causing 48.2 per cent of the outbreaks. Salmonella enterica was involved in 34 per cent of them. The most frequent serovar was typhimurium. Foods involved were: cheese in 29.3 per cent of the cases; cakes in 15.5 per cent; cooked meat in 15.1 per cent; milk in 13.8 per cent; and fish and seafood in 7.0 per cent of the cases. Since the number of studied incidents represents only a small proportion of all the outbreaks occurring in the country, the constant exchange of information among the laboratories which work on the problem and the promotion of the health care team are necessary in order to improve the epidemiologic surveillance systems and the study and prevention of food-borne disease and food poisoning outbreaks. PMID- 8235892 TI - [Family influence in the metabolic control of the type-2 diabetic]. AB - This paper presents the results of the identification and association of certain family characteristics of type II diabetes patients in relation with its metabolic control in 121 families of tw o health centers of Guadalajara, Jalisco, Mexico in 1989. The attitude an d information about of the sickness, and the complications and therapeutics were evaluated through psychosocial evaluations. The majority of the family members showed a negative attitude and a profound understanding of the disease process. Only a 16.5 per cent of the patients were under control, and found a strong association between lack of control and nuclear family involved in the last cycles of this type of unit: independence and retirement. PMID- 8235893 TI - [Community participation in health at the Instituto Mexicano del Seguro Social in Jalisco]. AB - This paper describes the achievements and problems of a community participation model in health care, developed by the Mexican Social Security Institute since 1985. The health team involved between 10 and 20 percent of the personnel due to the slow adoption this program requires in new of the cultural change it implies. The community's answer has been satisfactory, as it can be seen by the ware of 100 neighbours and school committees, and around the 2,000 housewives and school students who are been working as promoters. This figures stand monthly. The self care groups on chronic diseases constitute another form of participation. The community activities are progressively focused on causes of health and disease, embracing our 70 per cent of health promotion and disease prevention actions. The work realized by the community over cause one third of that realized by the preventive medicine personnel. This health community participation model represents a valuable and viable alternative. PMID- 8235894 TI - [Research in tropical medicine and primary health care in Peru]. AB - Tropical medicine's fundamental task is to improve health in the tropics. By adopting primary health care strategies, it satisfies the real needs of the population while doing research, improving its effectiveness and social impact. We illustrate this with some examples drawn from our experience, where this potentiation is evident. A sanitary dermatology study, based on health auxiliaries and promoters, encompassed a whole jungle province, with 68,977 km2 and 103,681 inhabitants. It resulted in an excellent relationship with the populations, and findings of significance for early diagnosis and control of hanseniasis and other diseases. It also facilitated an extension of activities to include the entire Amazonian Region, with specific concentration on training of the health personnel. Clinico-epidemiological studies on leishmaniasis in Andean valleys incorporated activities of sanitary education, health care, aspects of community development, etc., and extended into other geographic areas. Migrant workers from high-altitude communities in Cusco who have been to the jungle and acquired cutaneous or mucocutaneous leishmaniasis formed Patient Associations. The latter now receive support for their health and development needs from health authorities and many institutions; our Institute contributes with improved therapeutic procedures and further epidemiologic studies to orient preventive and control measures. PMID- 8235895 TI - [Errors in the certification of deaths from cancer and the limitations for interpreting the site of origin]. AB - The aim of this study is to assess errors in the filling-out of death certificates and difficulties for disclosing the origin of neoplasms. This study was carried out in Tijuana, Mexico. Death certificates containing a diagnosis of cancer were selected from all death certificates issued from January 1 to December 31, 1989. The following variables were identified: age, sex, place of residence and cancer site. Using these criteria, we found that 74.9 per cent of death certificates had been correctly filled out and 21.5 per cent were errors. It was seen that the main cancer sites causing mortality were the lungs, breast, cervicouterine and stomach. Cancer in unknown sites was registered in 20.5 per cent of all certificates. Implications of these results are analyzed in the light of mortality trends by causes and risk factors. We suggest that training courses are given to physicians so that they fill out the certificates properly. PMID- 8235896 TI - [A strategy for preventing health injuries due to observing the solar eclipse in Mexico]. AB - A total solar eclipse was watched by almost [corrected] 50 million people in the Mexican Republic on July 11, 1991. Cases of solar retinitis, which can even lead to permanent loss of visual function, have been reported in the international literature. The institutions of the National Health System employed a strategy for the prevention of risks and health damages caused by direct observation of the phenomenon, which implied the elaboration of a technical norm concerning the manufacturing of sun filters, the diffusion of preventive actions through the use of mass media, the detection and prompt care of cases and the establishment of an ophthalmologic care system and epidemiological surveillance system. The result was the detection of 21 moderate cases of solar retinitis, all of which recovered their full visual function after four months. The present article reports the implemented actions and the details of the cases. PMID- 8235897 TI - [An evaluation of a symposium via satellite on alcoholism and drug dependence]. AB - Results of a test given to participants in a symposium on alcoholism and drug abuse are presented. The symposium was broadcast via satellite simultaneously to five cities in Mexico, and included 8 pretaped panels covering topics on alcoholism and drug abuse. The methodology used for broadcasting the symposium allowed the interactive exchange of information between expert lecturers and participants. The quantitative and qualitative evaluation used the pretest posttest design. Most of the participants were physicians (28.9%) followed by psychologists (25.7%) and social workers (18.1%). The global cognitive change among participants was 6 per cent. Almost 77 per cent of participants had scores between 51 and 70 points (over a possible maximum score of 100) in the pre evaluation test, and 76.4 per cent had scores between 61 and 80 points in the postevaluation test. Health professionals with 1-3 years of experience had the largest change in scores (9%), followed by those with 3 to 5 years experience (8%). Professionals with 5 to 10 years of experience had a change of 5 per cent. Physicians showed the greatest cognitive change (7%) followed by psychologists and social workers with 5 per cent change. In the qualitative evaluation, 87.6 per cent of participants thought that the educational method used was "Excellent" of "Good". With respect to the satellite transmission, 79.4 per cent of participants thought it was "Excellent" or "Good". It is concluded that this type of educational events are useful in reaching health professionals who otherwise would not have access to specialized and updated information.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235898 TI - [The prevention of sexually transmitted diseases and AIDS by the use of the condom]. AB - This paper deals with the last literature and examines the most relevant studies of recent years related to condom effectiveness in preventing sexually transmitted diseases (STD), with experimental, epidemiological and seroconversion models. On the basis of these studies, it can be stated that the efficacy of condoms is close to 100 per cent when used correctly and consistently. Condom breakage has been reported in 0.5 per cent of vaginal intercourse and 3.6 to 10 per cent of anal intercourse. Most studies refer low failure rates due to manufacturing defects. Considering all constraints involved in masculine condom use and their implications for women, a female condom has been developed recently. It is not enough to give information or to make condoms available, if all the elements involved in the enhancement or failure of its use are not considered. Sexual education programs are the fundamental strategy in preventing STD and pregnancy. PMID- 8235899 TI - [Tetraethyl lead. 1925]. PMID- 8235900 TI - [The use of digital luminescence radiography in pediatric thoracic diagnosis]. PMID- 8235901 TI - [The first clinical experiences with spiral-CT-angiography of the abdomen]. PMID- 8235902 TI - [Initial results of the use of a new film-screen system in the UV region]. PMID- 8235903 TI - [The use of a "rapid sequence" in the diagnosis of tumors in the head and neck region]. PMID- 8235904 TI - [Rapid coloration and a fan-shaped puncture technique for improving the fine needle aspiration biopsy of palpable lesions of the female breast]. PMID- 8235905 TI - [Diagnostic and therapeutic radiologic interventions on the bile ducts]. PMID- 8235906 TI - [Magnetic resonance tomography of malignant peripheral soft tissue tumors]. PMID- 8235907 TI - [Anomalies of the inferior vena cava as pictured by computed tomography]. PMID- 8235908 TI - [The situational stress of radiation oncology patients--the possibilities of psychosocial support]. PMID- 8235909 TI - [Seven mysteries in rheumatoid arthritis]. PMID- 8235910 TI - [Two cases of acute progressive interstitial pneumonia associated with dermatomyositis--clinical features and immunological disorders]. AB - Some patients of dermatomyositis (DM) with interstitial pneumonia (IP) have common clinical features. Clinical features of these patients are acute onset, very poor prognosis and that patients have fever, arthritis, typical skin rash, mild myositis and show low ratio of CPK/LDH, low incidence of antinuclear antibody (ANA) appearance, low inflammatory signs. We experienced two cases of this category of DM with IP and examined immunological aspects. Case 1. A 52-year old woman was admitted in June 1, 1990 with a one-month history of arthralgia and a ten-days history of fever, skin rash, myalgia and dyspnea on exertion. On examination she had Gottron's papules on her fingers, erythema on back, bilateral elbows and legs, proximal muscle weakness and arthritis. Fine crackles were audible in the lower lung fields. Laboratory data included CPK 200 IU/l, ALD 3.2 IU/l, LDH 805 IU/l. Analysis of bronchoalveolar lavage fluid (BALF) revealed increased cellularity with lymphocytosis. She was treated with oral corticosteroid (CS), pulse CS, cyclosporin A. Inspite of these therapies, she died of progressive respiratory insufficiency in July 10, 1990. Case 2. A 23-year old woman was admitted in April 1, 1991, with a two-month history of arthralgia and a one-month history of fever, skin rash, stomatitis, alopecia. On examination she had Gottron's papules on her fingers, erythema on malar, bilateral elbows and legs, arthritis and stomatitis. Laboratory data included CPK 97 IU/l, ALD 8.5 IU/l, LDH 779 IU/l. She began experiencing dry cough and dyspnea on exertion in May 1991. Analysis of BALF revealed increased cellularity with lymphocytosis. She was treated with oral corticosteroid(CS), pulse CS, pulse cyclophosphamide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235911 TI - [Bronchoalveolar lavage fluid analysis in rheumatoid arthritis]. AB - In order to clarify the pulmonary involvement in rheumatoid arthritis (RA), bronchoalveolar lavage (BAL) was performed in 52 RA patients, in conjunction with pulmonary function test (PFT) and high-resolution CT (HRCT) of the chest. The results were as follows: 1. Out of nine RA patients with no evidence of lung involvement on HRCT, six (66.7%) showed abnormal differentials of the cells obtained through BAL. 2. RA patients with advanced interstitial lung disease on HRCT had significantly higher BAL fluid (BALF) cellular concentrations, increased percentage of BALF neutrophils and decreased BALF CD 4/CD 8 ratio, as compared with those with no or mild lung involvement. 3. In the non-smoking RA patients who also demonstrated no obstructive patterns on PFT, positive correlations were observed between %VC and the BALF CD4/CD 8 ratio (R = 0.481) as well as the BALF CD4+Leu8- (helper T) cell ratio (R = 0.497). On the other hand, a negative correlation was detected between %VC and the BALF CD8+CD11b- (cytotoxic T) cell ratio (R = -0.533). 4. In those with the increased percentage of BALF eosinophils and/or with increased BALF CD4/CD8 ratio, V25/Ht was depressed significantly, as compared with other patients. Likewise, those with severely depressed value of V25/Ht tended to have significantly elevated BALF CD4/CD8 ratio, as compared with other patients with normal V25/Ht value. Thus, through these detailed studies of BAL cellular differentials and BALF lymphocyte subsets, the diversity of pulmonary involvement was well demonstrated in RA. PMID- 8235912 TI - [Infection after implant arthroplasty]. AB - From 1982 to 1990, ten patients (twelve joints) who developed deep infection after total joint replacement were analysed. An average follow-up period after the surgical management was two years and eight months (range, one year and seven months to three years and ten months). Synovectomy and closed irrigation were carried out in seven of ten patients without waiting the results of cultures. In six of seven patients, the infection was successfully eradicated without removal of the components. Antibiotics were administered orally for two years after the closed irrigation. There was no recurrence. In four of ten patients, the components had to be removed. In one of four patients, synovectomy and closed irrigation failed, and arthrodesis was carried out. In three patients, loosening of the components were radiographically noticed. PMID- 8235913 TI - [Immunoglobulin decrease accompanying adverse reactions with bucillamine]. AB - Forty-two RA patients treated with Bucillamine (B) were divided into two groups with (n = 26) and without (n = 16) adverse reactions, and IgG, M, A and total protein concentration x globulin ratio (Ig) before and after B were compared. IgG did not differ before B administration, but was significantly lower in the adverse reactions group after B, and in each group it was significantly lower after B treatment with a much greater decrease demonstrated in the adverse reactions group. IgA before B did not differ either between the two groups or between before and after B administration in the non-adverse reaction group, but was significantly lower only after B in the adverse reaction group. IgM did not differ either before or after B treatment between the two groups, and was significantly lower following B administration in both groups. Ig showed results similar to IgA. In the adverse reaction group, IgG, A, M and Ig recovered following suspension of administration. It is concluded that the adverse reaction brought on by B administration might occur with the decrease in immunoglobulin. PMID- 8235914 TI - [A case of systemic lupus erythematosus who developed nodular shadows in the lung in parallel with the disease activity]. AB - A 35-year old woman was admitted to our hospital in April 1991 due to the appearance two nodular shadows on chest X-ray. The patient's past medical history was as follows; In 1986, she developed polyarthralgia, facial eruptions and Raynaud's phenomenon which was treated with prednisolone. In 1988, investigations revealed proteinuria where by the diagnosis of SLE complicated with lupus nephritis was established. Subsequently, treatment with cyclophosphamide was carried out. However, in January 1991, there was further increase in urine proteins in accompaniment with systemic eruptions. In addition, laboratory findings showed an increasing titre of anti-DNA antibody and decreasing serum compliments. In April 1991, as indicated before, a chest X-ray revealed nodular shadows in the right upper and lower lung fields, therefore, a transbronchial lung biopsy was performed. The histology revealed inflammatory infiltrates in the bronchoalveolar tissue, fibrosis of the septa- and exudates in the alveolar space. These findings suggested fibrosing alveolitis. It was true considered that these lesions in the lung were associated with the exacerbation of SLE. In view of this, the dose of prednisolone was increased to 60 mg/day. Consequently, the nodular shadows regressed and the clinical course of SLE improved. It is well appreciated that some patients with SLE develop variable pulmonary involvement, however, this case is of great interest due to the fact that multiple nodular shadows on chest X-ray are very rare in SLE. PMID- 8235915 TI - [Giant cell arteritis and polymyalgia rheumatica presenting as subclavian artery obstruction]. AB - Giant cell arteritis (GCA) and polymyalgia rheumatica (PMR) are common diseases in the elderly. The arteritis usually affects medium sized vessels, but large vessel involvement can also occur leading to arm claudication, bruits, loss of pulses and pallor of the upper extremities. The differential diagnosis of large vessel arteritis includes atherosclerosis and Takayasu's disease. Atherosclerosis, which affects patients of similar age to GCA is usually confined to the lower limbs and can be differentiated on the basis of the clinical setting and investigations such as the ESR, arteriography and temporal artery biopsy. Takayasu's arteritis' although histologically and arteriographically indistinguishable from GCA, is predominantly a disease of young women. A patient is described who presented with upper limb ischemia. A clinical examination revealed absence of right radial pulses and presence of murmurs at level of the carotids. The blood pressure was unrecordable in the upper right limb. The ESR was 102 mm/hr and the C-reactive protein was 11.66 mg/dl. A selective arteriography of the aortic arch and its branches revealed a right subclavian artery obstruction with good collateral circulation and a left subclavian artery stenosis. The biopsy of left temporal artery showed a typical GCA in acute stage. Treatment with prednisolone 30 mg/day was started and four weeks later, the ESR had fallen to normal. In addition this case confirms that PMR implies a systemic arteritis. PMID- 8235916 TI - [Renal and cerebral infarctions in a patient with systemic lupus erythematosus without antiphospholipid antibodies]. AB - Renal artery infarction is a very rare complication in patients with systemic lupus erythematosus (SLE), even in patients with antiphospholipid syndrome which often causes thromboembolism: Renal infarctions have only been reported in 4 SLE patients with antiphospholipid antibodies (aPL). Here we report a case of SLE without aPL who accompanied by renal and cerebral infarctions. A 42-year old Japanese woman with 8 year history of SLE manifested by arthralgia, central nervous system symptoms, positive-antinuclear and anti-DNA antibodies was admitted to our hospital for the treatment of progressive lupus nephritis. Physical examinations revealed hypertension (130-160/80-110 mmHg) without pitting pretibial edema. Laboratory evaluations showed proteinuria (3.7 g/day), normal serum creatinine level (0.9 mg/dl), low serum albumin level (2.3 g/dl) and high cholesterol level (317 mg/dl). Old cerebral infarctions were recognized by magnetic resonance imaging. However, hematological and immunological studies revealed that this case has neither a prolonged activated partial thromboplastin time, lupus anticoagulant nor anticardiolipin antibodies. Prednisolone was increased from 30 mg/every other day to 30 mg/day, and oral azathioprine, 50 mg/day, was started for the treatment of lupus nephritis. On the 11th day, she suddenly complained severe abdominal pain, which gradually localized on the right side. Computed tomography of the kidney suggested right renal infarctions, and arteriography of right renal artery confirmed both an obstruction of the ventral branch and a narrowing of the dorsal branch of right renal artery. No intra cardiac thrombus was demonstrated by echocardiography. Following to the treatment with fibrinolytic agent and anticoagulant, her symptoms have improved.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235917 TI - [Causes of loosening after total hip replacement and revision surgery]. PMID- 8235918 TI - [Study on Japan Rheumatism Association diagnostic criteria for early rheumatoid arthritis. 1. Application of the American Rheumatism Association diagnostic criteria to Japanese patients with early rheumatoid arthritis]. AB - The Clinical Research Committee of the Japan Rheumatism Association is planning to formulate the diagnostic criteria for early rheumatoid arthritis (RA). We have analysed 123 contemporaneously and consecutively registered patients with early RA and 61 control subjects with rheumatic diseases other than RA (non-RA). As the first step of the works, these 184 patients were applied to the criteria of ARA in 1985 and 1987 for the sensitivity, specificity and accuracy of each items. Consequently, we found that the two ARA criteria were not sufficiently sensitive as the diagnostic criteria for patients studied in this work. Therefore it would be necessary to formulate a new diagnostic criteria for the patients with early rheumatoid arthritis. PMID- 8235919 TI - Comparison of DNA-probe test and culture for the detection of Neisseria gonorrhoeae in genital samples. AB - BACKGROUND AND OBJECTIVES: The nonisotopic DNA probe assay (PACE 2) was evaluated for the detection of Neisseria gonorrhoeae in urethral and endocervical specimens and compared quantitatively and qualitatively using recommended culture methods. GOAL OF THIS STUDY: To evaluate whether culture methods could be replaced by the DNA probe assay for the diagnosis of N. gonorrhoeae. STUDY DESIGN: A total of 502 men and women were studied, including 42 non-registered and 132 legally registered prostitutes and 115 male and 213 female STD patients. RESULTS: The overall concordance of culture and the PACE 2 assay was 98.4%. Of the tested persons, 9.6% were positive for N. gonorrhoeae by culture technique compared with 11.2% using the DNA probe assay. All samples that were positive in culture were positive in the Gen-Probe test. Of samples positive in Gen-Probe, 14.3% (8/56) were negative in culture. Three of these results were determined false-positives and five were true positives when evaluated by further analysis. The sensitivity and specificity were 90.6% and 100%, for the culture technique, and 100% and 99.3% for the DNA probe assay, respectively, when compared with true positive and negative results. CONCLUSION: The data indicate that the DNA probe assay serves as a suitable screening and diagnostic test for the diagnosis of N. gonorrhoeae in symptomatic and in asymptomatic men and women. When the performance of culture technique is not possible or unreliable, the DNA probe assay can be used for testing those at high risk for gonorrhea. In borderline cases with a low value of RLU, the DNA test should be confirmed to avoid false positive results, especially in women. PMID- 8235920 TI - An in vitro human epithelial cell culture system for studying the pathogenesis of Chlamydia trachomatis. PMID- 8235921 TI - Influence of syphilis in hepatitis B transmission in a cohort of female prostitutes. AB - BACKGROUND AND OBJECTIVES. Prostitutes are a greater risk for hepatitis B virus (HBV) infection than the general population. We studied the influence of age and time as prostitute on HBV infection. We also examined the relationship between syphilis and HBV infection in a cohort of female prostitutes. STUDY DESIGN. The presence of hepatitis B surface antigen (HBsAg), antibodies to hepatitis B core antigen (anti-HBc), antibodies to hepatitis D virus (anti-HD) and treponemal antibodies (FTA-ABS) were determined in 368 prostitutes, of whom 147 were submitted to medical and serological follow-up every six months to evaluate the influence of syphilis in the transmission of hepatitis B. RESULTS AND CONCLUSION. The prevalence of HBsAg was 4.6%, of anti-HBc 31.2%, anti-HD 0.5% and FTA-ABS 35.0%. There was a statistical association between the presence of treponemal antibodies and anti-HBc (P = 0.022). The cohort study performed shows that the accumulated incidence of HBV infection in the FTA-ABS positive prostitutes (24.6%) was significantly higher than that of the FTA-ABS negative group (9.7%) (RR = 2.544; P = 0.034). Our results indicate that syphilis could facilitate the heterosexual transmission of HBV infection. PMID- 8235922 TI - Comparative evaluation of ofloxacin and metronidazole in the treatment of bacterial vaginosis. AB - BACKGROUND AND OBJECTIVES: The epidemiology and etiology of bacterial vaginosis is poorly understood. Many antimicrobial agents have been studied and suggested as treatment alternatives for this infection. Fluoroquinolones have in vitro activity against some of the organisms associated with bacterial vaginosis and may provide another treatment option. STUDY DESIGN: Women who presented with symptomatic bacterial vaginosis were randomly assigned to receive either oral ofloxacin 300 mg twice a day or oral metronidazole 500 mg twice a day for seven days. Twenty-seven of the 31 patients who were enrolled in this double-blinded study were evaluable. RESULTS: Fourteen women received ofloxacin. At follow-up, four had no residual symptoms, seven noted a reduction in symptoms, and three had no change in symptoms. Eight (57%) of the 14 women who received ofloxacin had secretions at follow-up that satisfied the criteria for the diagnosis of bacterial vaginosis. Thirteen women received metronidazole. Nine had no residual symptoms and four noted a reduction in symptoms. Eleven (85%) of the 13 women had normal vaginal secretions and two (15%) had secretions that were improved. None of the women who received metronidazole had secretions at follow-up that met the criteria for the diagnosis of bacterial vaginosis. CONCLUSION: We conclude that ofloxacin is less effective than metronidazole in the treatment of bacterial vaginosis. PMID- 8235923 TI - Vaginal infections in diabetic women: is empiric antifungal therapy appropriate? AB - BACKGROUND: Diabetes mellitus is reported to predispose women to vaginal candidiasis and hence patients attending busy diabetic clinics are often treated empirically with antifungal agents for genital symptoms. GOAL OF THIS STUDY: To investigate the etiology of vaginal infections in diabetic women and to determine appropriateness of empiric antifungal therapy for symptomatic women. DESIGN: Prospective study performed on consecutive patients attending two diabetic clinics. SETTING: Diabetic clinics serving developing communities belonging to two ethnic groups. STUDY POPULATION: Two hundred and one women (101 African and 100 Indian) comprising 90 women with symptoms of pruritus vulvae and/or vaginal discharge and 111 asymptomatic women. RESULTS: Candidiasis (50% vs. 24%; P = 0.012) and bacterial vaginosis (28% vs. 8%; P = 0.017) occurred significantly more frequently in symptomatic African women compared to asymptomatic African women. Among Indian women bacterial vaginosis (28% vs. 5%; P = 0.026) occurred more frequently in symptomatic women. The prevalence of trichomoniasis was similar in symptomatic and asymptomatic women of both ethnic groups. Mixed vaginal infections were detected significantly more often in African compared to Indian women (24% vs. 5%; P = 0.03). CONCLUSION: In diabetic women with genital symptoms, an attempt at diagnosis should be made prior to commencement of therapy. However, in busy clinics with overstressed facilities where investigations cannot be performed, the use of empiric antifungal therapy alone is not appropriate and consideration should be given to the use of an antifungal plus a nitroimidazole agent which would be effective for both trichomoniasis and bacterial vaginosis. PMID- 8235924 TI - Multicenter, comparative study of cefotaxime and ceftriaxone for treatment of uncomplicated gonorrhea. AB - BACKGROUND AND OBJECTIVES: Cefotaxime is a third-generation cephalosporin that has in vitro activity against Neisseria gonorrhoeae, including beta-lactamase producing strains. A single 1-g intramuscular dose is effective and is recommended by the Centers for Disease Control and Prevention as an alternative treatment for uncomplicated gonorrhea. GOAL OF THIS STUDY: This study was conducted to evaluate the efficacy and safety of a lower 500-mg dose of cefotaxime in the treatment of uncomplicated gonococcal infections. STUDY DESIGN: In a randomized multicenter study, patients who had uncomplicated gonorrhea were treated with 500 mg of cefotaxime or 250 mg of ceftriaxone. Both antibiotics were given intramuscularly. Efficacy and safety were assessed four to seven days following treatment. RESULTS: Six hundred thirteen patients were enrolled. Bacteriologic eradication rates for anogenital infection were 97.7% of the patients (213/218) in the cefotaxime group and 99.1% of the patients (221/223) in the ceftriaxone group (P = 0.243). Adverse events occurred in 4.2% and 7.5% of patients in the two groups, respectively. CONCLUSION: Cefotaxime 500 mg appears to be a safe and cost-effective alternative to ceftriaxone 250 mg for the treatment of uncomplicated gonorrhea. PMID- 8235925 TI - Determinants of genital human papillomavirus infection in low-risk women in Portland, Oregon. AB - OBJECTIVES: To confirm the risk factors for genital human papillomavirus (HPV) infection. GOAL OF THIS STUDY: To investigate risk factors for HPV detection independent of the correlated risk factors for cervical neoplasia in a low-risk population. STUDY DESIGN: HPV DNA was assessed among 483 cytologically normal women with no known history of cervical neoplasia. A cervicovaginal lavage was collected for HPV detection and typing using a PCR-based DNA amplification system. Information on risk factors of subjects was obtained through a questionnaire. RESULTS: HPV DNA was found in 17.7% of study women. On univariate analysis, factors associated with increasing HPV prevalence included younger age, fewer years of education, lower income, higher lifetime number of sex partners, lower age at first intercourse, nulliparity, oral contraceptive use, and current smoking. After statistical adjustment, we found younger age and higher number of sex partners were strongly and independently associated with higher HPV prevalence. We also observed increased HPV prevalence among women with lower levels of education and lower incomes. CONCLUSION: These findings and corroborative data from the companion reports in this issue of the journal support the sexual route of transmission of the virus. PMID- 8235926 TI - Determinants of genital human papillomavirus infection in low-income women in Washington, D.C. AB - OBJECTIVES: To confirm the risk factors for genital human papillomavirus (HPV) infection. GOAL OF THIS STUDY: To investigate risk factors for HPV detection independent of the correlated risk factors for cervical neoplasia, in a high-risk population. STUDY DESIGN: We investigated 404 cytologically normal women attending medical assistance clinics in the Washington, D.C. area. Risk factor information was obtained and a cervicovaginal lavage was collected and used for HPV detection and typing by a PCR-based technique. RESULTS: The point prevalence of HPV was 33.7%. This contrasts with the 17.7% and 44.3% observed in the companion reports published in this issue of the journal. HPV prevalence decreased with age and increased with greater numbers of sexual partners. Moreover, more recent sexual behavior was a better predictor of current HPV detection than lifetime number of sexual partners. Numbers of pregnancies and current pregnancy were positively associated with HPV prevalence and there was an indication that current oral contraceptive users had a higher prevalence of HPV compared to never users. Smoking was not associated with increased HPV prevalence. CONCLUSION: Findings support the sexual route of transmission of HPV and confirm the association of HPV detection with age, suggesting the host's ability to clear infection or the virus' ability to become latent. PMID- 8235927 TI - Determinants of genital human papillomavirus infection among cytologically normal women attending the University of New Mexico student health center. AB - OBJECTIVES: To confirm the risk factors for genital human papillomavirus (HPV) infection. GOAL OF THIS STUDY: To investigate risk factors for HPV detection apart from the correlated risk factors for cervical neoplasia. STUDY DESIGN: Cervical human papillomavirus (HPV) DNA was assessed in 357 cytologically normal women attending the University of New Mexico student health center. Cervical swab samples were obtained for HPV DNA detection and typing using a PCR-based DNA amplification system. Possible determinants of cervical HPV were examined including age, ethnicity, history of sexually transmitted disease, oral contraceptive use, smoking, age at first intercourse, lifetime number of sex partners, marital status, and history of pregnancy. RESULTS: A 44.3% overall prevalence of cervical HPV was observed. On univariate analysis, factors associated with increasing HPV prevalence included higher lifetime number of sex partners and single marital status. After adjustment for potential confounding variables, we found that HPV prevalence increased with higher lifetime number of sexual partners. CONCLUSION: These findings, along with those from the companion reports in this issue of the journal, support the sexual route of transmission of the virus. PMID- 8235928 TI - Treatment of uncomplicated gonorrhea with single doses of 200 mg cefixime. AB - BACKGROUND: Single-dose cefixime 400 mg orally is effective in the treatment of uncomplicated gonorrhea. However, lower doses of cefixime have not been studied, and the minimum effective single-dose regimen may risk selecting resistant strains of Neisseria gonorrhoeae. Therefore, we studied the efficacy of a lower dose of cefixime. OBJECTIVE: To assess the efficacy of 200 mg cefixime in the treatment of uncomplicated gonorrhea. METHODS: One hundred twenty-five patients (106 men, 19 women) with presumptive gonorrhea were treated with single doses of cefixime 200 mg orally in an open non-comparative study. Follow-up visits were scheduled for four to seven days after treatment. RESULTS: Genital and rectal gonorrhea were eradicated in 93 (95%, CI95 90.5-99.2%) of 98 patients who were culture-positive at enrollment and returned for follow-up. Treatment was effective in 78 (95%) of 82 men with urethral infection and 15 (94%) of 16 women with anogenital infection. Two of three pharyngeal infections also were eradicated. Persistent infection was not associated with resistance to cefixime, penicillin, or tetracycline. CONCLUSIONS: Cefixime 200 mg in a single dose has substantial efficacy in the treatment of uncomplicated gonorrhea, enhancing confidence that use of 400 mg cefixime for gonorrhea has a low risk of selecting gonococci with clinically significant antibiotic resistance. However, the 200 mg dose should not be used for routine treatment. PMID- 8235929 TI - Risk factors for sexually-transmitted diseases among deployed U.S. military personnel. AB - BACKGROUND AND OBJECTIVES: Information regarding risk factors for STD transmission is needed to assist in designing and evaluating prevention and control programs for US military populations. GOAL OF THIS STUDY: To obtain STD risk factor data among deployed U.S. military personnel. STUDY DESIGN: A questionnaire survey was administered to military personnel deployed aboard ship for six months to South America, West Africa, and the Mediterranean during 1989 1991. RESULTS: Among 1,744 male subjects (mean age, 23 years; 71% white; 96% enlisted), 49% reported prior sexual contact with a prostitute and 22% reported a history of a STD before deployment. During the subsequent six-month deployment, 42% reported sexual contact with a prostitute, 10% reported inconsistent use of condoms, and 10% acquired a new STD. By logistic regression analysis, sexual contact with a prostitute during deployment was independently associated with young age, nonwhite race/ethnicity, and being unmarried or divorced; inconsistent use of condoms was associated with Hispanic race/ethnicity. CONCLUSION: These data indicate that deployed U.S. military personnel frequently engage in high risk sexual behavior and that there is a continued need for comprehensive and culturally-sensitive STD prevention programs. PMID- 8235930 TI - [Role of aldosterone in essential arterial hypertension complicated by chronic renal insufficiency]. AB - The following parameters were studied in a group of patients suffering from essential arterial hypertension complicated by chronic renal insufficiency (CRI), mean age = 64.31 +/- 1.84, with creatinine clearance (CrC) ranging between 30 and 60 ml/min: blood pressure (systolic and diastolic arterial pressure), heart rate, plasmatic renin activity (PRA), plasma levels of aldosterone (ALDO) both in clino and orthostatism, as well as some metabolic parameters. All parameters were compared with those in a group of age- and sex-matched patients with slight or moderate essential arterial hypertension. Before starting the study all patients completed a wash-out period of one week to annual the effects of other drugs which might interfere with the RAA system. PRA levels were within the norm, whereas plasma levels of ALDO were high both in clino- and orthostatism. ALDO levels were also found to be inversely correlated with those of CrC. From these data it emerges that hyperaldosteronism, as observed in these patients with CRI, is a relatively reliable marker of the extent of CRI and may occur independently of the activation of the RAA system, given that other factors, such as orthostatic stimulation, alterations in the acid-base equilibrium, and the degree of aldosterone hepatic and urinary clearance, contribute to its pathogenesis. PMID- 8235931 TI - [Interaction between active metabolites of vitamin D3 and blood aluminum in chronic uremic patients on dialysis]. AB - In the present study, the authors show that 1, 25 (OH)2 D3 given to hemodialyzed patients taking Al (OH)3, increased their plasma concentrations of Al. Two mechanism can explain this increase: increased intestinal absorption or decreased tissue storage of Al. PMID- 8235932 TI - [Prevention of postoperative scrotal edema]. AB - The authors describe the technique of scrotal elevation and compression for the prevention of postoperative scrotal hematoma. This technique was used in 30 patients undergoing hydrocelectomy, spermatocelectomy or epididymectomy. There have been no untoward effects associated with this technique and this technique is applicable to a wide variety of scrotal procedures. The method described is simple, rapid to perform, comfortable for the patients and most effective to prevent postoperative complications in patients undergoing a scrotal operation. PMID- 8235933 TI - [Multilocular benign cystic nephroma. Case report and review of the literature]. AB - The authors report a case of multilocular cystic nephroma. According to the literature this is a rare lesion, which may be diagnosed any time during life, showing the same prevalence in childhood as in adults. Multilocular cystic nephroma is usually an unilateral lesion. Aetiology is unknown and preoperative ultrasonography and roentgenographic studies frequently led to misdiagnosis. Surgical excision of the cyst, with kidney sparing, is the therapy of choice, however nephrectomy is advised when intraoperative biopsy is ambiguous. PMID- 8235934 TI - [2 cases of unexpected anatomopathological discovery of a leiomyoma of the urogenital system]. AB - The authors report their recent experience in two cases of leiomyoma of the genitourinary tract not only because of relatively low incidence of this neoplasm in such sites, but especially since in the first case a previous urothelial lesion had led to the performance of a cystectomy and bilateral uretero-ileo cutaneostomy, which led us to attribute the later urethral lesion also to a transitional-type neoplastic site; in the second case, symptomatologic evolution, palpation and scrotal echography aroused suspicion of testicular neoplasm and only the observation during surgical operation made it possible to decide for a testis-preserving therapy enabled by the reassuring extemporaneous histological report. PMID- 8235935 TI - [Nephropexy using the Aglietti procedure. Case reports]. AB - The paper reports two cases of 3rd degree nephroptosis which were treated surgically using the technique described by Prof. Aglietti. Both cases were women suffering from associated pathologies. The need to distinguish between symptoms related to renal ptosis and those caused by different disorders required an accurate clinical and instrumental study to be performed in both patients. Only after the concomitant pathologies had been treated could the patients undergo nephropexy. The paper gives a brief description of the technique. The authors discuss the indications for surgery and analyse the results. PMID- 8235936 TI - [Sarcomatous degeneration of a giant scrotal lipoma. Presentation of a clinical case]. AB - This paper presents a case of giant scrotal lipoma with areas of sarcomatous degeneration which were evident on anatomico-pathological examination; the authors considered it worth-while reporting this case given that giant lipomas with sarcomatous degenerations are rare. It is important to underline that in comparison to other malignant tumours of the scrotum the frequency of liposarcomas is very low. In addition to the liposarcoma reported here, at the level of the scrotum it is worth recalling neurofibrosarcoma and fibrosarcoma and some rare sarcomas of the spermatic cord among the malignant sarcomas of sarcomatous origin reported in the literature. Difficulties may arise from a diagnostic point of view due to the scarcity of symptoms. Among the most frequent symptoms is the presence of a hard mass on palpation of the scrotum; the volume of this mass may increase very rapidly. It is important to note that this tumour may sometimes originate as malignant and then degenerate in toto or in some isolated areas. Instrumental tests include abdominal radiography without the use of contrast medium in order to exclude the presence of herniated abdominal viscera at the level of the scrotum. Ultrasonography may also be extremely useful since it is capable of providing information regarding the presence, inside the scrotal sack, of liquid retention or solid forms. At all events histological diagnosis is carried out in the majority of cases following the removal of the operated part. The exeresis of the tumour therefore becomes both diagnostic and therapeutic. Surgery consists in the simple exeresis of the neoplasia; in some cases orchiectomy is required in addition to exeresis. Some authors also recommend the dissection of inguinal lymph nodes following extemporary anatomo pathological tests. In other cases when histological tests confirm the malignant degeneration with a high degree of biological aggressiveness, the patient has to undergo postoperative radiotherapy and/or chemotherapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235937 TI - [Nephrogenic adenoma of the bladder associated with urinary tuberculosis. Case report]. AB - Nephrogenic adenoma is an uncommon benign lesion of the urinary tract, that histologically is characterised by glandular-like aspects resembling the distal part of the nephron. It is usually associated with antecedent inflammation, surgical procedures or other injuries. Personal experience with one additional case nephrogenic adenoma of the bladder in a patient with urinary tract tuberculosis is presented. PMID- 8235938 TI - [Standardization of methods for studying fibrinolysis]. PMID- 8235939 TI - [Splenic lymphoma with circulating villous lymphocytes. Study of 6 patients]. AB - PURPOSE: Splenic lymphoma with circulating villous lymphocytes (SLCVL) is a rare chronic B-type lymphoproliferative disorder, few series having been reported thus far. The major clinical, cytological, immunophenotypic and ultrastructural features, as well as the course and treatment of six cases seen in three hospitals of the Barcelona area in six years are reported herein. METHODS: The criteria of Melo et al were followed for the diagnosis of SLCVL. The clinical manifestations at onset and the morphology of lymphocytes from blood and bone marrow (aspiration and biopsy samples) were analysed. The samples were subjected to cytochemical (acid phosphatase and L-tartaric acid inhibition), immunophenotypic (alkaline immunophosphatase and immunofluorescence) and ultrastructural studies. The histologic study of the spleen of those patients who underwent splenectomy was also performed. RESULTS: The median age of the series was 63 years (range: 45-86). Five of the patients were women. Anaemia was the commonest clinical finding at onset, and splenomegaly was found in all cases. Villous lymphocytes were found in both peripheral blood, 3% to 85%, and bone marrow, 31% to 70%; they showed diffuse positivity to acid phosphatase stain, this being inhibited in all cases by L-tartaric acid. None of the patients had associated monoclonal gammopathy. Bone-marrow biopsy was performed in five occasions, showing nodular infiltrative pattern in three cases, interstitial in one and diffuse pattern in another. Nodular infiltration was seen in the white pulp of spleen, with no involvement of the red pulp. The ultrastructural study of villous lymphocytes from peripheral blood or bone-marrow disclosed short, slim villi with narrow bases, no lamellar ribosomal complexes being found. Mature B type lymphocyte proliferation was present in all instances, showing positivity for surface immunoglobulin; monoclonal antibody CD25 was negative in all cases. Three patients underwent splenectomy, and the remainders received no treatment. The expected survival at three years was 80%. CONCLUSIONS: SLCVL is a rare B-cell chronic lymphoproliferative disorder with recognised morphologic and immunophenotypic characteristics that make it distinguishable from other B-type lymphoproliferative diseases especially hairy cell leukaemia. Its clinical course is chronic and it has good prognosis. Response to splenectomy is usually favourable, although remission of the disease is not achieved. PMID- 8235940 TI - [Potentiation of the antiplatelet effect of dipyridamole and aspirin by erythrocytes. Study under flow conditions]. AB - PURPOSE: To assess, by means of a morphometric method, the interaction of platelets with subendothelium, as induced by perfusion with the Baumgartner system, and to compare the antiplatelet effect of acetylsalicylic acid (ASA) and dipyridamole (DIP) and the combination of both drugs. MATERIAL AND METHODS: Blood samples were drawn from healthy volunteers and platelet-rich plasma specimens were prepared at a concentration of 4 x 10(8)/mL. Abdominal aorta segments were obtained from New Zealand rabbits, they being deprived of endothelium by alpha chymotrypsin. Red cells and platelets were separately or conjointly treated on reconstituted whole blood using ASA and DIP at therapeutic doses, separately or in association. Perfusion experiences were carried out in Baumgartner's annular chamber. Upon finishing the studies, the aorta segments were morphometrically evaluated by an optic analysis system attached to a computer with an automatic image recognition programme. The statistical analysis was performed with Student's t test, differences between groups of p < 0.05 being assigned significance. RESULTS: Treatment with the association ASA-DIP was the only one to significantly reduce interaction, as both the size of aggregates and the extent of the covered surface decreased when whole blood was treated, and only the covered surface did when red cells were separately treated. When treating only platelets, the reduction of aggregates had no significance. CONCLUSION: The synergism of ASA and DIP in antiplatelet therapy was confirmed, as well as the important enhancing effect exerted by red cells. PMID- 8235941 TI - [Initial iron deposits in idiopathic myelofibrosis. Analysis of 20 patients]. AB - PURPOSE: To analyse the status of iron deposits at diagnosis in patients with idiopathic myelofibrosis (IM). PATIENTS AND METHODS: The study included twenty consecutive patients with IM. Haemoglobin, MCV, serum iron and serum ferritin were determined in each case. Perls stain was used to evaluate iron deposits in bone marrow samples attained by aspiration or biopsy. An attempt was made to correlate the semiquantitative estimation of bone marrow iron with the peripheral blood values connected with iron deposits. RESULTS: Anaemia was present in 15 patients, but in most cases MCV and serum iron were normal. Iron deposits in bone marrow aspirate could be evaluated only in one patient as in the remainders no material was obtained. The semiquantitative estimation of iron in bone marrow biopsy samples showed diminished or absent deposits in 15 cases (75%). In contrast, none of the patients had decreased serum ferritin levels. No correlation was found between bone marrow iron and the peripheral blood values related to iron deposits. CONCLUSION: Most IM patients have decreased bone marrow iron deposits at diagnosis. However, as in chronic myelogenous leukaemia and in essential thrombocytopenia, such finding does not mean a true iron deficiency status. PMID- 8235942 TI - [Invasive aspergillosis in patients with malignant hemopathies. Usefulness of direct and serologic diagnosis]. AB - PURPOSE: To evaluate the efficacy of different diagnostic methods for invasive aspergillosis (IA) in patients with haematologic neoplasms. PATIENTS: Eight patients were evaluated for this study (5 non-Hodgkin's lymphomas, 2 acute leukaemias, and 1 chronic myelogenous leukaemia in blastic crisis). Depending on the clinical characteristics of each case, one or more diagnostic procedures were used: (a) microscopic observation of fresh samples from sputum, urine sediment, or fine-needle aspirate; (b) culture in Sabouraud agar, glucose-gentamicin medium of the pathologic products; (c) serologic techniques for detecting anti Aspergillus spp antibodies and circulating antigen; (d) histologic examination of biopsy or necropsy specimens. RESULTS: The combination of microbiological methods (i.e., direct observation, culture, serology) provided the IA diagnosis in the 8 cases, this being confirmed by histologic observation in 5 instances (3 of them in necropsy studies). The observation of fresh samples allowed us to identify hyphae on different locations, although the culture of such material not always was positive. Seroconversion for Aspergillus spp was seen in 4 patients, whereas the antigen detection was negative in the 5 patients in whom this was attempted. CONCLUSIONS: (1) These findings suggest that different microbiological techniques must be combined for IA diagnosis when suspected in a severely compromised patient. (2) Direct microscopic observation may be of help, as it is a simple method allowing faster results than culture. (3) Serologic techniques, especially circulating antigen detection, are not fully developed. PMID- 8235943 TI - [Frequency of platelet alloantigens in the Spanish population]. AB - PURPOSE: The aim of this study is to investigate the distribution and incidence of platelet alloantigens in Spain and to compare this frequency with that found in other populations. MATERIAL AND METHODS: Five hundred blood donors were phenotyped for the HPA-1a (Zwa), HPA-3a (Bak(a)), HPA-4a (Yukb/Pen(a)) y HPA-4b (Yuk(a)). One hundred of these donors were also phenotyped for the HPA-5a (Br(a)) antigen. Furthermore, we also phenotyped a group of 50 black individuals from a community of west African immigrants who live in the province of Barcelona. The typing of all the antigens except those of the HPA-5 system was carried out using the technique of Indirect Immunofluorescence. To type the antigens of the HPA-5 system the MAIPA technique was employed. RESULTS AND CONCLUSIONS: The incidence of the different platelet specific antigens in the Spanish population is, in general, the same as that detected in the other European countries and in the white population of the USA. In Europe and North America, the HPA-1a antigen is the most important of all owing to its involvement in alloimmune clinical disorders. In Asia, this role seems to be played by the HPA-4b antigen. Moreover, polymorphism for the HPA-1a allele does not seem to exist. The incidence of the HPA-5b (Br(a)) antigen in Spain, which is higher than that for France and Germany, suggests a high involvement of this antigen in our country in different alloimmune processes. The preliminary data obtained in a group of black west Africans suggest the absence on polymorphism for the HPA-1a allele and that the incidence of the remaining antigens is comparable to that of the Spanish population. PMID- 8235944 TI - [Neutrophil chemotactic dysfunction in multitransfused thalassemia patients]. AB - PURPOSE: To evaluate the chemotactic capability of neutrophils in thalassaemic patients under poly-transfusion regimens and in thalassaemia carriers. PATIENTS AND METHODS: Twenty-one patients in multi-transfusion regimen diagnosed in the Ricardo Gutierrez Children Hospital were studied. Of them, 17 had thalassaemia major, 3 S/beta thalassaemia and one sickle-cell anaemia. Twenty-one normal subjects comprised a control group. Chemotaxis was evaluated by two methods, namely, migration under agarose layer and in microchemotaxis chamber under stimulation with N-formyl-methionyl-n-phenylalanine at optimal concentrations of 10(-5) M and 10(-6) M, respectively. RESULTS: In thalassaemia major patients, directed mobility of neutrophil assessed by both methods was significantly decreased with regard to the normal controls, whereas random mobility was preserved. The four patients under poly-transfusion who had not thalassaemia major showed the same neutrophil defect. On the contrary, chemotaxis and random mobility of the neutrophils from thalassaemia carriers (thalassaemia minor) were similar to those of the normal controls. CONCLUSIONS: These results suggest that the defect found in the patients might be caused by transfusion overload. PMID- 8235945 TI - [Clinical profile and course of paroxysmal nocturnal hemoglobinuria]. AB - PURPOSE: To evaluate the clinical characteristics, laboratory findings, course, treatment and survival of a series of patients with PNH. PATIENTS AND METHODS: The clinical records of 21 patients diagnosed of PNH in the La Fe University Hospital between 1970 and July 1991 were revised. Positivity to Ham's and sucrose tests was the major diagnostic criterion. Haemolysis was studied by means of reticulocyte count, unconjugated bilirubin, LDH levels, haptoglobin, serum iron, and haemosiderinuria. The survival analysis was performed in accordance with Kaplan and Meier. RESULTS: The median age in the group was 38 years (range: 18-72 years) and the M/F ratio was 11/10. The commonest symptoms at onset were weakness (76%), dark urine (47%), jaundice (33%), and purpura (24%). All the patients had anaemia, six of them (28%) presenting with aplastic anaemia. The complications most frequently seen included thrombosis (documented in 7 cases, clinically suspected in 6 others), infection (6 cases) and haemorrhage (6 cases). One patient developed aplastic anaemia after 16 years of follow-up, and another one had AML. None of the patients with PNH-associated aplastic anaemia developed thrombosis. The incidence of severe cytopenia was: Hb < 80 g/L, 62%; platelet count < or = 20 x 10(9)/L, 29%, and white cell count < or = 2 x 10(9)/L, 14%. The haemolysis-related findings were as follows: unconjugated hyperbilirubinemia, 100%; haemosiderinuria, 87%; decreased haptoglobin, 80%. Transfusion support consisted of washed red cells (total amount, 1,684 units) and platelets (137 units). A female patient with anticoagulant therapy developed haemolysis after non-isogroup plasma transfusion. Five patients required no transfusions. One patient was subjected to splenectomy and other underwent a successful bone-marrow transplant. The cause of death was related to PNH in three of the five patients who had died when this study was finished (mesenteric thrombosis, subcapsular haematoma of the liver and AML). The actuarial survival at 10 years was 68%. CONCLUSIONS: 1) The incidence of PNH is very low in our experience, excessive delay in diagnosis being the rule, as in other reported series. 2) Early anti thrombotic treatment should be carried out in PNH whenever severe thrombocytopenia is not associated. 3) Allogeneic BMT is the only curative treatment. PMID- 8235946 TI - [Colony-stimulating factors and interleukins. Regulators of hematopoietic production]. PMID- 8235947 TI - [Transient appearance of anti-erythrocyte autoantibodies during Rh alloimmunization]. AB - Transient appearance of anti-erythrocyte autoantibodies was demonstrated in two women in the first stages of Rh alloimmunization. Any mimicking antibodies, or concurrent autoimmune haemolytic anaemia could be reasonably discarded in both patients. These facts are discussed on the basis of polyclonal activation followed by clonal selection driven by the antigen, as a mechanism of the humoral immune response. Investigation of such cases seems of great interest in subjects with recent alloimmunization in order to assess the incidence of this phenomenon and the antigen systems involved. PMID- 8235948 TI - [Transfusion policy toward a patient with an anti-Cartwright antibody (anti Yta)]. AB - We have observed one patient with hepatic cirrhosis showing on her antibody screening an anti-Yta. We studied the 51 Cr-labelled erythrocyte survival using either Yt (a+) and autologous blood red cells. She was transfused two Yt (a+) red blood cell concentrates without any adverse reaction. PMID- 8235949 TI - [In vitro granulopoietic cultures. Analysis using monoclonal antibodies]. AB - Four monoclonal antibodies were applied, using an immunoenzymatic technique, the alkaline phosphatase anti-alkaline phosphatase, in granulopoietic cultures in vitro. In this manner, granulopoietic growth at tenth day of culture has been defined as positive for myeloid markers (CD15, CD11b) and negative for macrophagic (CD68) and proliferating cells (Ki 67) markers. PMID- 8235950 TI - [Cytogenetic study of 4 patients diagnosed with splenic B-cell lymphoma with villous circulating cells]. AB - We present the cytogenetic study of 4 patients diagnosed of splenic B-cell lymphoma with villous lymphocytes. In such patients, it is noteworthy the chromosomal abnormality, del(7)(q32), found in two cases (in one as a sole chromosomal abnormality). PMID- 8235951 TI - [Transient reticulocyte increase in patients undergoing bone marrow transplantation after erythrocyte concentrate transfusion]. PMID- 8235952 TI - [Treatment of anemia in myelodysplastic syndromes with recombinant human erythropoietin]. PMID- 8235953 TI - [Comparative study of the determination of hemoglobin (capillary and venous blood) in blood donors]. PMID- 8235954 TI - [Hemoglobinopathies and thalassemias in the Hindu settlement established in the Canary Islands]. PMID- 8235955 TI - Aims and opportunities in clinical anatomy. PMID- 8235956 TI - Anatomic basis for use of a gracilis muscle flap. AB - The aim of this study was to specify certain anatomic features of the gracilis m. with a view to the use of muscular or myocutaneous flaps. It was based on dissection of 84 gracilis muscles in 42 subjects as well as selective injection of the main pedicle of 20 muscles. This established the following points: 1) The arterial supply is abundant, consisting of several pedicles reaching the muscle on its deep aspect. The main neurovascular pedicle arises from the deep vessels of the thigh, via either the a. of the adductors (73%), the medial circumflex a. (19.2%) or as a double supply from both arteries (7.7%); 2) The cutaneous vascularisation over the gracilis m., derived from the solitary main pedicle, is inconstant. In 20 injections, it was satisfactory in 11 cases, poor in 5 and absent in 4; 3) The distal tendon of the gracilis m. is closely related to the posterior branch of the saphenous n. to the leg, which it crosses in an elongated X; 4) A simple method of calculation based on the distance between the upper border of the pubis and the medial femoral epicondyle allows quite precise determination of the point of entry of the main pedicle into the gracilis m. 5) Complete dissection of the main pedicle adds to the available length of the muscle flap. PMID- 8235957 TI - The trochlear nerve: anatomy by microdissection. AB - This work is based on the microscopic study of 30 trochlear nerve trunks (15 heads). In 17 cases, the trunk arose from two nerve bundles, in 8 cases from one bundle, and for the other 5 nerves, three or four bundles. The mean total length of the trochlear nerve was 86 mm. The nerve may be separated into the 3 following parts: infratentorial, intracavernous, intraorbital. In all 30 cases studied, the first part of the nerve was infratentorial, thus leading us to suggest the term "infratentorial part" for this segment of the nerve. In 27 cases, contact was found with the superior cerebellar artery, in the infratentorial part. In the intracavernous part of ten nerves we found two rami tentorii and in eight cases fibers were exchanged with the ophthalmic nerve. In the orbit, 18 trochlear nerves crossed the posterior ethmoidal artery. 23 trochlear nerves ended on the medial face of the superior oblique muscle. The remaining 7 ended at the superior border of the muscle. PMID- 8235958 TI - Microsurgical anatomy of the inferior alveolar neurovascular plexus. AB - Microdissection of the cadaveric human inferior neurovascular bundle showed that the inferior alveolar nerve after its origin from the mandibular division forms what can only be described as a plexus. The main trunk of the inferior alveolar nerve exhibits a branching structure reminiscent of the brachial plexus of the upper limb. This plexus-like nature is compounded by delicate interweaving of the inferior alveolar artery. Study of external carotid angiograms showed a general decrease in size of the inferior alveolar artery with increase in age. PMID- 8235959 TI - Densitometric analysis of the paranasal sinuses. AB - The knowledge of weak points in the osseous structure of the skull is of great importance for endonasal operations. For this reason the anatomy of the skull is accurately described in serial sections. Serial sections of 1.5 mm thickness are evaluated by a computerized image analyzing system, which allows qualitative as well as quantitative statements on the structure of the paranasal sinuses and their neighbourhood. The bone density in the frontobasal area of the skull as well as of the ethmoid cells is presented separately. The anatomy of the sphenoid sinus is described by densitometric and gross techniques. PMID- 8235960 TI - Arteries to the great and second toes based on three-dimensional analysis of 100 cadaveric feet. AB - Three-dimensional arteriography was used to analyse the arterial supply of the great and second toes of 100 cadaveric feet down to the microsurgical level. This information will aid in successful composite tissue transfers of these toes to the hand. The arterial blood supply of the great toe came principally from the first dorsal metatarsal a. (78%) and the first plantar metatarsal a. (22%), and secondarily from the medial tarsal aa. and the three terminal branches of the medial plantar a. For the second toe, the first dorsal metatarsal a. (78%) and the first plantar metatarsal a. (22%) supplied blood from the medial side, and the second dorsal metatarsal a. (78%) and the second plantar metatarsal a. (22%) supplied blood from the lateral side. Seven arterial patterns were found in the interdigital web space. The so-called general pattern was seen in the first web space in 65% of the feet examined. In the second web space it was found in 85%. The first intermetatarsal space sometimes contained a large artery arising directly from the dorsalis pedis or first proximal perforating a. as well as the first dorsal and first plantar metatarsal arteries. In this space, arterial patterns were classified into 4 types and 9 subtypes based on the origins and proximal courses of these arteries. The so-called standard pattern was found in only 19% of the feet, while an arterial pattern with a common proximal trunk on the plantar side for the first dorsal and first plantar metatarsal aa. was found most frequently (46%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8235961 TI - An anatomic study on the Martin-Gruber anastomosis. AB - The incidence of Martin-Gruber anastomosis was studied. Anatomical dissection of 108 cadaveric arms was performed. The Martin-Gruber anastomosis was found in 23 arms. This anastomosis was classified into four types: Type Ia: Communication between the anterior interosseous and the ulnar nn. (n = 13), Type Ib: Communication between the median and ulnar nn. (n = 1), Type II: Communication between muscular branches of the flexor digitorum profundus m. (n = 8), Type III: Combination of Type Ia or Ib and Type II (n = 1). The incidence of Martin-Gruber anastomosis was found to be significantly higher in the present study than in previous studies. PMID- 8235962 TI - Arteria thyroidea ima arising from the brachiocephalic trunk with bilateral absence of inferior thyroid arteries: a case report. AB - The anomalous first parts of the left and right subclavian aa. had no inferior thyroid aa. in the neck region. The thyroidea ima a. was found to arise from the brachiocephalic trunk, and bifurcated into two branches almost immediately after its origin. These branches ascended in front of the trachea and entered the bases of the right and left lobes of the thyroid gland. The left vertebral a. arose from the aortic arch in the superior mediastinum. The possible existence of this anomaly is important for parathyroid localization studies, in neck surgery and especially in tracheostomy. PMID- 8235963 TI - An abnormal pattern of blood vessels in the retroperitoneal space with a duplicated inferior vena cava in an adult: a case report. AB - During dissection a duplicated inferior vena cava (IVC) was found in an 82 year old male. The IVCs were connected with each other by two anastomoses. The course of the other retroperitoneal veins also exhibited some alterations: Just below the diaphragm the azygos and the right ascending lumbar v. ran into the right IVC separately; the hemiazygos and the left ascending lumbar vv. joined before entering the left IVC. The testicular vv. ended normally on the right side in the right IVC, on the left side in the left renal vein. The embryologic basis of the pattern of these vessels is discussed and a short overview of the literature given. In the present case there were also some alterations in the course of the renal and testicular aa. PMID- 8235964 TI - Unilateral duplication of the vestibular aqueduct. AB - A case of unilateral duplication of the vestibular aqueduct is presented. This very rare abnormality can be explained either by an excessive induction by the rhombencephalon or by an abnormal complete division of the endolymphatic duct into utricular and saccular parts. PMID- 8235965 TI - Lymphatic drainage of the esophagus in the adult. PMID- 8235966 TI - Anatomic basis of mandibular reconstruction by free vascularised fibular graft. AB - The free osteofasciocutaneous fibular transplant is a direct development of the free fibular bone graft used from 1983 in reconstructive surgery of the long bones. In 1989, Hidalgo was the first to publish a method of reconstruction of the mandible by composite free fibular transplants. The aim of this study was to specify the anatomy of the osteofascio-cutaneous fibular graft and particularly of the vessels to the skin. The sensory innervation of this transplant is described. PMID- 8235967 TI - Risks and cost-effectiveness of sublaminar wiring in posterior fusion of cervical spine trauma. AB - A report on the use of sublaminar wiring in the fusions of 34 patients with cervical spine injuries is given. No neurologic deficits were incurred in the use of this technique. There were no wire failures nor clinically significant complications. When compared with other forms of instrumentation to achieve similar results, the use of sublaminar wire is the most cost-effective. PMID- 8235968 TI - Effect of nimodipine or methylprednisolone on recovery from acute experimental spinal cord injury in rats. AB - The purpose of the present study was to examine the behavioral, electrophysiologic, and anatomic responses to nimodipine or methylprednisolone treatment of acute experimental spinal cord injury. Four groups of rats were injured at T1 by compressing the cord with a 52-g clip for 1 minute. The treatments were begun 15 minutes after injury, and the animals were observed thereafter for 8 weeks. Nimodipine 0.02 mg/kg/h intravenously (iv) for 8 hours with adjuvant albumen volume expansion, followed by 20 mg/kg nimodipine enterally three times per day for 7 days, produced a moderately better composite score comprising four endpoint parameters than the other treatments which consisted of nimodipine iv for 8 hours only, methylprednisolone 30 mg/kg iv bolus followed by 5.4 mg/kg/h iv for 8 hours, or control. PMID- 8235969 TI - Long-term outcome in surgically treated spina bifida cystica. AB - This report describes the long-term operative outcome of 72 patients with spina bifida cystica. The period of follow-up was between 4 and 20 years. In our series, 17 patients died, with the mortality rate increasing as the lesions were more rostral. All cases involving only a meningocele are living without handicap. The cases of spina bifida cystica with hydrocephalus had higher morbidity and mortality when compared to those without hydrocephalus. We conclude that the rostro-caudal location, the content of the sac, and whether there is associated hydrocephalus are important factors influencing the long-term prognosis of spina bifida cystica. PMID- 8235970 TI - A joint protocol for the neurosurgical and neuroradiologic treatment of cerebral arteriovenous malformations: indications, technique, and results in 76 cases. AB - We report on a group of 76 patients who had been treated during a period in which preoperative embolization using interventional neuroradiologic technique was standard procedure. During this time, 20 consecutive patients were operated on without embolization, eight patients were treated with embolization only, and 48 patients were operated on after embolization. In 35 cases surgery immediately followed the last embolization procedure. All arteriovenous malformations (AVMs) were classified and the outcome analyzed according to Spetzler [J Neurosurg 1986; 65: 476-83]. Those lesions treated with a combination of embolization and surgery had higher Spetzler grading than those that had been operated without previous embolization. The overall recovery rate was 82.9%. In 7.9% of the cases the AVMs were not totally extirpated. The overall complication rate was 9.2%. It is concluded that preoperative embolization facilitates surgery and reduces the risk of severe morbidity and mortality, especially in high-grade lesions. In addition to the clinical results, the application and usefulness of Ethibloc as an embolizing agent is reported. PMID- 8235971 TI - Postradiation volume changes in gamma unit-treated cerebral arteriovenous malformations. AB - Postradiation changes in angiographically determined nidus volume were quantitatively studied in 22 arteriovenous malformation (AVM) cases treated by gamma unit radiosurgery. The postradiosurgical decrease was statistically significant by post-treatment year 2 (p < .05). In children, AVMs tended to be obliterated more quickly than in adults. Volume reduction was more rapid in nidi receiving 25 Gy or more than in those receiving less than 25 Gy (p < .01). However, there were no significant differences in nidus volume decrease between these two dose groups at the second or third postradiosurgical year. Dose response curves were obtained 1, 2, and 3 years following treatment. PMID- 8235972 TI - Vertebral hemangioma late results of retrograde embolization--stabilization with methyl methacrylate in two cases. AB - In two cases of hemangioma of the D7 and L1 vertebral bodies intraoperative posterior filling of the angioma was described. The technique allowed a conveniently bloodless operation, which resulted in stabilization. No other stabilization was necessary. Six months after the procedure an increase of vertebral body density was seen radiologically. PMID- 8235973 TI - Solitary intracranial chondroma of the convexity dura: case report. AB - We present a rare case of chondroma originated from the dura mater of the cerebral convexity in a 16-year-old girl. Radiologic findings are reported with emphasis on computed tomography and magnetic resonance imaging scans, and histogenesis is briefly discussed. PMID- 8235974 TI - Self-inflicted orbital and intracranial injury with a retained foreign body, associated with psychotic depression: case report and review. AB - Reports of intracranial self-mutilation by psychotic individuals are associated with severe mental disorders, criminality, or both. We describe a psychotically depressed male who drove a ballpoint pen through his right medial canthus and into his intracranial compartment. The patient developed a cavernous sinus syndrome and a traumatic dissection of the cavernous portion of the carotid artery. The pen was removed intraoperatively. Postoperatively, the patient was placed on a course of broad-spectrum antibiotics, antidepressants, and antipsychotic medications, and he has received long-term psychiatric follow-up. The literature related to these unusual cases is reviewed, and relevant surgical, medical, and psychiatric aspects of treatment are discussed. PMID- 8235975 TI - Giant cervical dumbbell-shaped neurinoma treated by transuncodiscal approach. AB - We treated a case of giant dumbbell-shaped neurinoma extending from the C-3 to C 6 spinal canal and causing significant compression of the cord posteriorly. Total removal of the tumor was performed by transuncodiscal approach. The usefulness of this approach will be detailed in this paper. PMID- 8235976 TI - Cranial epidural fibrous tumor associated with hyperostosis: a case report. AB - This report describes an unusual frontal epidural fibrous tumor in an adult woman. There had been no prior surgical procedure, radiation therapy, or significant head trauma. Microscopically, the lesion consisted of hypocellular, sparsely vascular, dense fibrous tissue. The fibrous mass was accompanied by hyperostosis of the frontal bone. The clinical and pathological characteristics of this lesion are illustrated. The differential diagnosis is discussed with reference to the literature regarding intracranial fibrous lesions. We are unaware of previous report of similar lesions in this location. PMID- 8235977 TI - Hydromyelia associated with a posterior fossa cyst. AB - There are rare reports of children with hydromyelia in association with arachnoid cysts at the foramen of Magendie, and these cases have uniformly been associated with hydrocephalus. We report a case of a 45-year-old woman with a posterior fossa cyst associated with hydromyelia and normal ventricles. This was successfully treated with a cystoperitoneal shunt. We believe this unusual condition is of interest in elucidating potential mechanisms of hydromyelia. PMID- 8235978 TI - Intracranial meningeal melanocytoma: a report of two cases and a review of the literature. AB - Two rare cases of intracranial meningeal melanocytoma are reported with clinical, radiologic, and pathologic findings. In one of these the diagnosis was established by ultrastructural studies that showed premelanosomes and melanosomes in nonarachnoid-derived tumor cells. The other case was diagnosed retrospectively after a very long survival and is still alive 35 years after total excision and radiotherapy. The occurrence of these tumors in the cerebellopontine angle is rare as seen in our case. PMID- 8235979 TI - Fibrous dysplasia of the clivus. AB - The case of a 37-year-old man with monostotic fibrous dysplasia of the clivus is reported. This study includes clinical presentation, radiologic appearance, surgical findings, and pathology of this lesion. The significance of magnetic resonance imaging and management of this uncommon clival lesion are discussed. PMID- 8235980 TI - [Editor's foreword on the concept of Strahlentherapie und Onkologie]. PMID- 8235981 TI - [Radiotherapy in adults with localized supradiaphragmatic stages CS I/II of Hodgkin's disease. 2. The special aspects of radiotherapy: dosage, fractionation, focal volumes, risks, conclusions]. AB - For patients with Morbus Hodgkin and CS I/II of the low risk group and primary radiotherapy recommended treatment fields are: regional field for isolated high cervical involvement, mantle field for isolated mediastinal involvement and extended mantle field for the other patients. Omission of the infradiaphragmatic irradiation volume for PS I/II may be regarded as an advantage, which must be compared with the risk of a staging laparotomy, whereas the low risk of undertreatment of a small part of patients with CS I/II PS III probably does not outweigh the risk of the laparotomy. Equal efficacy of chemotherapy alone for these patients has not been proven sufficiently and important questions concerning long-term risks are unanswered. Ongoing studies will show, whether combinations with reduced chemotherapy or other types of chemotherapy and local radiotherapy are superior. Details of the mantle field borders and blocking are described. In most patients with adjuvant radiotherapy after complete remission after chemotherapy, the recommended target volume includes only the regions with proven involvement before chemotherapy. Details of the mediastinal treatment volume for patients with adjuvant radiotherapy after chemotherapy for bulky mediastinal disease are given. According to some recent analysis of a large body of dose-effect data, the recommended target doses in primary irradiation are between 36 and 40 Gy for regions with proven involvement and between 30 and 36 Gy for electively treated regions. The recommended target dose per fraction is between 1.5 and 1.8 Gy and less than 2 Gy in various critical tissues. According to the recent recommendations, the maximal total doses in mantle field radiotherapy to the spinal cord should be 38 Gy for radiotherapy alone and 36 Gy for radiotherapy combined with chemotherapy. The maximal total dose to the whole heart should be 15 Gy and for the other parts between 30 and 35 Gy. After chemotherapy with MOPP oder MOPP-like regimes, there is a cumulative risk of leukaemia between 2.2 and 11.9%. After radiotherapy alone, there is only a very low risk of leukaemia after radiotherapy and chemotherapy to the risk after chemotherapy. Most long-term studies show an increased risk of solid second malignancies associated with radiotherapy with a relative risk of approximately two. In the analyzed studies, the cumulative risk of solid second malignancy after seven to 15 years is between 7 and 11.2% after radiotherapy, between 7 and 11.7% after chemotherapy and between 7 and 11.7% after radiotherapy and chemotherapy.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8235982 TI - Comparison of early glottic and supraglottic carcinoma treated with conventional fractionation of radiotherapy. AB - There were a few reports to compare the treatment results of glottic and supraglottic carcinoma. We investigated the difference of glottic and supraglottic carcinoma. From 1967 through 1985, 338 cases with early glottic (T1: 210, T2: 52) and supraglottic carcinoma (T1: 34, T2: 42) were treated with the conventional fractionation of radiation and evaluated the tumor response during radiation at the Department of Radiology, Osaka University Hospital. The ten-year actuarial survival rates of T1, T2 glottic and T1 and T2 supraglottic carcinoma were 63%, 64%, 58% and 46%, the corresponding ten-year cause-specific survival rates 92%, 89%, 88% and 78%, and the corresponding ten-year local control rates 76%, 67%, 69% and 59%, respectively. Tumor clearance rates of T1 and T2 glottic and T1 and T2 supraglottic carcinoma at 40 Gy were 68%, 40%, 35% and 24%, respectively. The local control rate of the tumor which disappeared at 40 Gy was 84%, that persisted at 40 Gy and disappeared at 60 Gy 69%, and that persisted at 60 Gy 33%. The local control rate and the tumor clearance rate of T1 glottic carcinoma were the highest among early laryngeal carcinoma, and those of T2 glottic carcinoma were the same as those of T1 supraglottic cases. The local control rate, the cause-specific survival rate, and the actuarial survival rate of T2 supraglottic were significantly lower than other cases. PMID- 8235983 TI - [A comparative consideration of para-aortic fields within the framework of the German Hodgkin's Disease Study]. AB - In 33 patients of the multicenter German Hodgkin's Lymphoma Study Group the field borders of the paraaortic field were compared computer-assisted. It was seen that fields are chosen often too small or too large, though precise description of the fields is given in the protocol. In these patients the para-aortics were irradiated exclusively as extended field region. In addition to the above observation the clipping of the spleen pedicle provides the optimal help for correct positioning of this field. Overall a very low frequency of lymphatic clipping is observed, though clinical needs are obvious. PMID- 8235984 TI - Comparison of systemic radiotherapy with I-131-labeled monoclonal antibody BW575/9 to external beam radiotherapy in human neuroblastoma xenografts. AB - The therapeutic effectiveness of external beam radiotherapy (XRT) and radioimmunotherapy (RIT) was investigated in a human neuroblastoma (SK-N-SH) xenotransplanted to nude mice. This tumor model seems especially suitable for comparison of the relative biological effectiveness of RIT vs. XRT, as--in contrast to most tumor models--it shows an unusually homogenous uptake of the labeled MAb, thus providing a homogenous intratumoral dose distribution. XRT was performed using single fractions of 800, 1600, 2000 and 2400 cGy and RIT was delivered by intravenous injection of 15, 19 and 26 MBq of the I-131-labeled monoclonal antibody (MAb) BW575/9. Therapeutic efficiency of the two radiation modalities was assessed in terms of tumor volume doubling time (VDT). Miniature thermoluminescent (mini-TLD) dosimetry and MIRD-based dose calculations were used to evaluate the absorbed doses delivered by RIT and to assess the degree of homogeneity of the dose distribution. RIT with 19 MBq of the I-131 BW575/9 delivered a tumor dose of 2820 cGy measured by TLD and resulted in a tumor VDT of 32 days (vs. one day in controls). An equivalent effect on VDT was achieved by a single fraction XRT of 1600 cGy. The relative efficiency of XRT compared with RIT (ratio of dose XRT/dose RIT required to give the same VDT) was 0.57. Application of 26 MBq of the MAb (= 3200 cGy) resulted in complete tumor regression after ten days as did XRT with 2400 cGy, corresponding to a relative efficiency of 0.75. PMID- 8235985 TI - [The mixed fractionated radiotherapy of R1H tumors in the rat using neutrons and photons]. AB - A number of experimental investigations has demonstrated, that tumor growth delay due to a mixed fractionated treatment schedule with photons and neutrons can be prolonged compared to pure photon treatment. That question after the mechanisms remains undissolved. In the presented study of the R1H tumor of the rat 30 fractions using 2 Gy of photons in 39 days are compared to results of mixed fractionated treatment schedules with neutrons and photons. Neither a prolonged net growth delay (NGD) was found nor does the sequence in which neutrons and photons are given influence the outcome when replacing photons by neutrons in twelve of 30 fractions (RBE 2.5). In contrast, hypofractionated schedules (five fractions/total dose 40 Gy/four days) have a considerable longer NGD when replacing two fractions of photons by neutrons (RBE 2.5) (p < 0.01). This could be explained if 8 Gy photons/fraction destroyed all euoxic tumor cells, a 24 hour interval was too short for reoxygenation and thus make hypoxia to a major problem in 8 Gy/fraction photon treatment. PMID- 8235986 TI - Estimate of normal tissue damage in treatment planning for stereotactic radiotherapy. AB - A personal computer (PC) system was developed to perform treatment planning for radiosurgery and stereotactic radiotherapy. These techniques of irradiation of the brain may be accomplished with a linear accelerator by performing several non coplanar arcs of a highly collimated beam focused at a fixed point. The PC system allows the acquisition, reconstruction and the visualization of the target volume from CT or MR images, and then it permits to calculate a three-dimensional (3-D) dose distribution due to small photon beams and to visualize it. The software calculates not only total dose distribution, administered fractionated or in single fraction, but also the NTD2 (normalized total dose) predicted to have a biological effect equivalent to the single irradiation. The choice of the best technique is supported by the dose volume histograms (DVH) calculation and by an estimate of complication probability to the brain normal tissue (NTCP). The algorithm for NTCP calculation is based on two models: the linear quadratic and the logistic. A comparison of three different dose calculations for a typical cerebral target volume is presented to demonstrate the system performances. PMID- 8235987 TI - Cure in a patient with multiple osseus metastases in non-small cell lung cancer: a case report. AB - PURPOSE: This case was reported to describe a case of cure in a 61-year old patient with squamous cell lung cancer and multiple extrathoracic metastasis. METHODS AND MATERIALS: A left upper lobectomy of lung for a squamous cell carcinoma was performed on a 61-year old man with curative intent. Four months later two osseus metastases were irradiated with Cobalt 60 up to 40 Gy. RESULTS: The two irradiated lesions showed continuously shrinkage as well as signs of recalcification. Eleven years later the patient shows clinically absolute well being and on CT there are no signs of recurrent disease of the lung or bone anymore. DISCUSSION: To our knowledge has nobody so far reported of a case of as squamous cell lung cancer which was operated and irradiated on thus resulting in cure. Further on the authors discuss that it might well be worthwhile to define subgroups in stage 4 non-small cell lung cancer (presence of extrathoracic metastases) which might benefit from a more aggressive treatment approach than pure palliation. PMID- 8235988 TI - Lung irradiation for Ewing's sarcoma with pulmonary metastases at diagnosis: results of the CESS-studies. AB - BACKGROUND: In the German CESS-studies, Ewing's sarcoma patients with pulmonary metastases at diagnosis were considered as candidates for additional bilateral lung irradiation. We have retrospectively analyzed the impact of radiotherapy on survival. MATERIAL AND METHODS: Out of 42 patients presenting with pulmonary metastases of Ewing's sarcoma between 1981 and June 1992, 30 were evaluable. One patient died of progressive disease prior to lung irradiation. The other patients had a complete radiographic remission either after chemotherapy (n = 25) or chemotherapy plus resection of lung metastases (n = 4). 22 patients received bilateral lung irradiation with doses of 12 to 21 Gy (once daily 1.5 Gy or twice daily 1.25 Gy), six had no further treatment and one had bone marrow transplantation for consolidation. RESULTS: Ten patients are in complete remission, nine of them had received lung irradiation and one had a complete resection of three lung metastases. Two patients are alive with disease, two have died from treatment-related complications and 16 have died from systemic disease with or without local relapse. NED-patients had received significant higher lung doses than relapsed patients p = 0.028. Moreover, a dose-response relationship was detectable: 1/6 patients without lung irradiation vs. 4/10 with 12 to 16 Gy vs. 5/6 with 18 to 21 Gy lung irradiation were in remission. CONCLUSION: In this retrospective analysis, lung irradiation was beneficial in terms of improved survival of patients with pulmonary metastases of Ewing's sarcoma. We actually recommend lung irradiation with 18 (to 20) Gy for pulmonary metastases of Ewing's sarcoma. PMID- 8235989 TI - [Radiotherapy in ductal in-situ breast carcinoma effective]. PMID- 8235990 TI - [Radiotherapy following quadrantectomy in breast carcinoma]. PMID- 8235992 TI - [Authorship in medical publications]. PMID- 8235991 TI - [Can a delay in radiotherapy be justified by postoperative chemotherapy in children under 3 with malignant brain tumors?]. PMID- 8235993 TI - Sources of error with use of Goldmann-type tonometers. AB - There are many sources of error in the use of Goldmann-type applanation tonometers. In clinically normal corneas hypofluorescence of the precorneal tear film, accommodation, the Valsalva maneuver and vertical gaze are preventable causes of large tonometric errors. Repeated tonometry may induce a decline in the intraocular pressure (IOP). Variations in the corneal resistance to indentation between eyes cause significant errors. The most significant cause of error in clinically abnormal eyes is corneal epithelial edema, which causes a marked underestimation of IOP. Measurements obtained with Goldmann-type tonometers can be used with confidence to monitor changes in the IOP of an individual, but should not be relied on to determine the absolute manometric pressure within an eye or to compare the IOPs in eyes of different individuals. PMID- 8235994 TI - Vertical diplopia. AB - An accurate clinical evaluation of vertical diplopia is predicated upon meticulous history-taking, observations regarding the presence and pattern of an anomalous head position, and the analysis of several subjective and objective tests of extraocular muscle function. To reach a final diagnosis with minimum risk and expense to the patient the examiner must be familiar with the neuroanatomy of the supranuclear and infranuclear pathways which control the actions of the vertically-acting extraocular muscles, the clinical methods and pitfalls of a number of clinical techniques which are used to identify an underacting extraocular muscle, and the hallmark characteristics of a supranuclear, infranuclear and restrictive ophthalmopathy. PMID- 8235995 TI - Visual loss and a suprasellar mass complicated by pregnancy. AB - A 43-year-old woman who was 12 weeks pregnant presented with a one-month history of visual loss in the right eye. Neuroimaging studies revealed multiple intracranial aneurysms, one of which compressed the right optic nerve. Her pregnancy raised many issues concerning her management. Evidence of aneurysm enlargement and the increased risk of aneurysm rupture as gestation progressed led to early, successful surgical intervention. PMID- 8235996 TI - Intracranial plasma cell granuloma presenting as an optic neuropathy. AB - A 40-year-old man presented with a left optic neuropathy. Magnetic resonance imaging demonstrated a contrast-enhancing mass along the course of the left trigeminal nerve and in the region of the left cavernous sinus with suprasellar extension. Preoperatively, he had a serum polyclonal gammopathy. Pathologic diagnosis was an intracranial plasma cell granuloma. The patient responded to high-dose steroids with resolution of his optic neuropathy, marked decrease in the size of the mass, and resolution of the serum polyclonal gammopathy. Intracranial plasma cell granuloma is a rare lesion; only 11 cases have been described in the literature. Almost half of the patients presented with vision loss as their chief complaint. PMID- 8235997 TI - Could do better: a curious clinician looks back--and forward. AB - A recently retired Scottish academic originally chose ophthalmology by a process of exclusion. Basic training in Glasgow and London was followed by "permanent" consultant appointments in Bristol, with a year as retina fellow at the Massachusetts Eye and Ear Infirmary, then London, then chairborne in Manchester and Edinburgh. Many opportunities for advances in ophthalmology were missed, but there were some successes: beta-blockers and steroid blockers in glaucoma, and gene mapping of one X-linked retinitis pigmentosa gene. Hypothesis-making is advocated by regarding no situation as static or sacrosanct, maintaining wide interests and collaborating with basic scientists. PMID- 8235998 TI - Fuchs' endothelial dystrophy of the cornea. AB - Fuchs' endothelial dystrophy of the cornea is a significant cause of corneal blindness in the United States. The disease is characterized by a slow, continuous loss of morphologically and physiologically altered endothelial cells, eventually leading to corneal edema. The endothelial cells synthesize a thickened Descemet's membrane with focal excrescences of altered basement membrane material (guttae). This review details the epidemiological, clinical, and laboratory data that have accumulated on Fuchs' dystrophy. Several hypotheses regarding the pathogenesis of Fuchs' dystrophy are discussed, including the possible influences of aberrant embryogenesis, hormones, and injury on the development of the disease. The current state of medical and surgical management is summarized, along with the future prospects for treatment. PMID- 8235999 TI - Basal cell carcinoma of the eyelid and periocular skin. AB - Basal cell carcinoma is the most common human malignancy. This locally invasive tumor rarely metastasizes, but can cause considerable morbidity when on the eyelids and periocular skin. The successful management of BCC requires an awareness of its broad clinical spectrum and knowledge of the risk factors associated with therapeutic failure. The optimal therapy for basal cell carcinoma is controversial. Treatment strategy is formulated on an individual basis, taking into account tumor size, anatomic location, and growth pattern. The immune status and life expectancy of the patient also must be considered. While complete eradication of the tumor is desirable, this can be challenging since basal cell carcinomas often extend beyond their apparent clinical margins. Since prevention is preferable to treatment, the public needs to be informed that the incidence of periocular skin cancers can be reduced with the longterm use of sunscreens, sunglasses, and hats with brims. PMID- 8236000 TI - Management of periocular basal cell carcinoma: Mohs' micrographic surgery versus radiotherapy. AB - In treating periocular basal cell carcinoma the goal is to achieve the most complete eradication of affected tissue while preserving as much healthy tissue as possible. In the first of these "Viewpoints" articles, Drs. Leshin and Yeatts discuss the success of Mohs' microsurgical technique (MMS) in both regards. Moreover, they note that MMS offers the additional advantage of placing all surgical and histological phases of tumor excision in the hands of one specialist, thus circumventing problems that may arise with procedures requiring the separate involvement of several specialists. In the second article, Drs. Anscher and Montana point out that modern radiotherapy techniques may offer adequate tumor control with less damage to surrounding tissues than is caused by surgical excision and reconstruction. With small lesions, radiotherapy has a success rate similar to that of surgery. With large lesions, the success rate is lower; however, if surgical excision would cause extensive disfigurement or functional damage, an initial trial of radiotherapy may be warranted, then followed by surgery, if necessary. In an editorial, Dr. Dutton summarizes the indications for and advantages and disadvantages of both techniques. PMID- 8236001 TI - The "pseudo-CSF" signal of orbital optic glioma on magnetic resonance imaging: a signature of neurofibromatosis. AB - A five-and-a-half-year-old boy with neurofibromatosis had bilateral orbital optic gliomas visible on magnetic resonance imaging. Both tumors displayed a double intensity signal characterized by a circumferential area of CSF-intensity tissue surrounding and sharply delimited from a central linear core of opposite signal intensity. The peripheral CSF-intensity signal in orbital optic glioma correlates with the histopathological finding of perineural arachnoidal gliomatosis and serves as a neuroradiologic marker for neurofibromatosis. PMID- 8236002 TI - Myasthenia gravis by another name: an elusive imposter. AB - A patient with procainamide-induced "pseudo" myasthenia gravis is presented. This clinical entity is discussed in terms of its clinical features, diagnosis, and treatment. Attention is directed to the importance of role of the ophthalmologist in making the diagnosis. PMID- 8236003 TI - Conjunctivitis of allergic origin: clinical presentation and differential diagnosis. AB - Conjunctivitis of allergic origin is associated with a diversity of clinical presentations. Common features include pronounced itching, a milky conjunctival appearance, a stringy or ropy discharge and papillary hypertrophy of the tarsal conjunctiva in severe cases, and a family history of allergy. The diagnosis should be based on thorough history-taking and careful ophthalmic examination, and, when necessary, confirmation by laboratory testing. An IgE-mediated immediate hypersensitivity mechanism is associated with most types of allergic conjunctivitis, although contact allergy is mediated by lymphocytes rather than antibody. Treatment is based on the diagnosis and severity of signs and symptoms. An array of medications is available to control symptoms, and the regimen of choice should be based on the response to milder forms of therapy and consideration of drug side effects. PMID- 8236004 TI - Double-masked, paired-comparison clinical study of ketorolac tromethamine 0.5% ophthalmic solution compared with placebo eyedrops in the treatment of seasonal allergic conjunctivitis. AB - In a multicenter, double-masked, clinical study, we compared the ocular safety and efficacy of ketorolac 0.5% ophthalmic solution with placebo in alleviating the signs and symptoms of allergic conjunctivitis. The study was conducted in 93 patients who received one drop of the appropriate treatments in each eye, four times a day, for seven days. Of these subjects, 74 were evaluated for efficacy. The principal clinical sign of allergic conjunctivitis, conjunctival inflammation, and six allergic symptoms (itching, swollen eyes, burning or stinging, discharge or tearing, foreign body sensation, and photophobia) were evaluated by the investigators at midweek (day 3 or 4) and at the end of the study (day 7 or 8). Ketorolac was superior to placebo in reducing conjunctival inflammation (p = 0.003) and itching (p = 0.020), the principal clinical symptom, at the final examination. In addition, ketorolac was favored over placebo in reducing the other five symptoms evaluated. On the day of final examination, overall therapeutic response evaluated by the investigators rated ketorolac as superior to placebo (p = 0.007). A significant placebo effect was noted in this study, as has been previously reported in clinical studies of allergic conjunctivitis patients. Results of this study demonstrate that 0.5% ketorolac ophthalmic solution used topically four times daily, for seven days, was effective in alleviating the principal sign and the symptoms associated with allergic conjunctivitis. PMID- 8236005 TI - Clinical evaluation of ketorolac tromethamine 0.5% ophthalmic solution for the treatment of seasonal allergic conjunctivitis. AB - We evaluated 148 patients with allergic conjunctivitis in a double-masked, paired comparison clinical trial comparing ketorolac 0.5% ophthalmic solution with vehicle. Patients received one drop of each study medication in preassigned eyes, four times a day, for seven days. Both treatments showed significant changes from baseline in the signs and symptoms associated with allergic conjunctivitis. Evaluations at the final visit (day 7 or 8) showed that ketorolac-treated eyes had a significant treatment response when compared to vehicle-treated eyes for conjunctival inflammation (p = 0.010), ocular itching (p = 0.006), swollen eyes (p = 0.002), discharge/tearing (p = 0.021), foreign body sensation (p = 0.035), and conjunctival injection (p = 0.016). Mean scores evaluating the overall therapeutic effect of the study treatments at the completion of the study were higher for ketorolac-treated eyes than for vehicle-treated eyes as rated by investigators (p = 0.004) and study patients (p < 0.001). Results of this study confirmed the trends of a previous study showing that ketorolac 0.5% ophthalmic solution applied topically is an effective therapy for allergic conjunctivitis. PMID- 8236006 TI - Differentiating conjunctivitis of diverse origins. AB - While symptoms can be very distressing, patients with conjunctivitis general maintain good vision and recover completely without permanent sequelae. The great majority of cases of conjunctivitis are infectious or allergic in origin. Seen with increasing frequency are external eye diseases related to contact lens wear or prolonged use of ophthalmic medications. The various forms of conjunctivitis are often not associated with pathognomonic features. A thorough history and ophthalmic examination often permit a presumptive diagnosis and initiation of empiric therapy. For example, a chronic bilateral conjunctivitis, characterized by itching and papillary hypertrophy, suggests an ocular allergy, most frequently the result of exposure to airborne allergens. However, a number of causes, including infections and hypersensitivity reactions, have the potential to threaten vision or produce marked conjunctival scarring which must be identified by the use of appropriate laboratory techniques, followed by specific therapy. Most bacterial and viral conjunctivitis are self-limited, but antimicrobial therapy for the former is advocated to shorten the course, improve patient comfort, prevent recurrence, avoid complications and limit spread to other individuals. PMID- 8236007 TI - Surgery for morbid obesity. PMID- 8236008 TI - Ten years of experience with patients with chronic active liver disease variceal bleeding: ablative versus selective decompressive therapy. AB - BACKGROUND: Variceal hemorrhage is an added major threat to survival in patients with chronic active liver disease (CALD). The hemodynamic consequences of surgical therapy and the continued activity of the underlying liver disease both contribute to the hepatic dysfunction and determine patient survival. METHODS: Two hundred and seventy two consecutive cases of (Child A or B) variceal bleeding with chronic hepatitis were surgically treated during a 10-year period. Histologic chronic active hepatitis (CAH) was documented in 160 (59%) patients, whereas chronic persistent hepatitis (CPH) was evident in 112 (41%). The applied surgical procedure was distal splenorenal shunt (DSRS) in 99 (36%) patients, splenectomy and gastroesophageal devascularization (SGD) in 108 (40%), and splenectomy with left gastric ligation (SLGL) in 65 (24%) patients. The preoperative data base obtained on these patients was matched comparing the three surgical modalities within each pathologic group (p > 0.05). RESULTS: The operative mortality was low among the patients with CAH (DSRS, 5.1%; SGD, 4.2%) with no deaths occurring in the CPH group. Among the individuals with CAH, recurrent variceal hemorrhage occurred significantly (p < 0.05) more often after SLGL (26%) and SGD (17%) than after DSRS (5%). Sclerotherapy rescued 93% (SGD) and 70% (SLGL) of the patients with rebleeding. DSRS significantly (p < 0.05) increased the risk of encephalopathy (28%) compared with SGD (4.2%) and SLGL (8%). The morbidity rates were quite low among the patients with CPH with no significant (p > 0.05) differences noted when the three surgical modalities were compared. Both groups experienced a significant (p < 0.05) increase in aspartate aminotransferase levels after the three procedures with a significant (p < 0.05) increase in bilirubin level occurring only after DSRS. The 5-year survival rate for the patients with variceal bleeding with CAH was 76% (DSRS), 73% (SGD), and 88% (SLGL). The leading causes of death were liver failure after DSRS (70%), variceal hemorrhage after SLGL (60%), and equally divided between septicemia (43%) and liver failure (43%) after SGD. The patients with CPH had a better 5 year survival of 89% (DSRS) and 100% (nonshunt operation). CONCLUSIONS: These data showed that (1) CALD is common among cases of variceal bleeding; (2) elective surgical treatment of variceal hemorrhage in patients with Child A or B CALD has a low operative mortality; (3) SLGL backed up by sclerotherapy is a better surgical alternative to either selective shunt or SGD in patients with active hepatitis, and (4) both DSRS and nonshunt operation are equally good surgical options for patients with CPH. PMID- 8236009 TI - Parathyroid carcinoma: diagnosis and clinical history. AB - BACKGROUND: Parathyroid carcinoma is often over-diagnosed based on histologic appearance alone. We limited the definition of the disease to patients with recurrence, metastases, or frank capsular invasion on histologic examination. METHODS: With these criteria, fourteen cases of parathyroid carcinoma seen at our institution from 1955 to the present were identified. RESULTS: All patients presented with hypercalcemia, and all deaths were due to hypercalcemia. Two patients have been free of disease after initial operation for 31 and 180 months. Six other patients had a prolonged course with a median survival more than 80 months. Two of these patients have undergone one reexcision each for local recurrence, and four have undergone multiple resections for local recurrence or metastases. These reoperations usually resulted in satisfactory, albeit temporary, control of hypercalcemia. Finally, six patients died of disease after an aggressive course, with a short median survival (47 months). Four of these patients were seen in the 1950s before an aggressive approach to metastatic disease was adopted. CONCLUSIONS: The types of clinical courses observed in this study may be more reflective of the varied biologic features of parathyroid carcinoma and the approach to recurrence than of the initial operations. For patients with recurrent or distant disease, an operation appears to prolong survival and palliate the symptoms of hypercalcemia. PMID- 8236010 TI - Long-term evaluation of the endorectal Soave operation performed for ulcerative colitis or polyposis in the pediatric patient. AB - BACKGROUND: A follow-up study was conducted to evaluate the late results of the operation that we have performed for ulcerative colitis and polyposis during the past 25 years. METHODS: Sixty-seven consecutive patients less than 21 years of age who underwent a standard operation for ulcerative colitis or polyposis performed by or under direct supervision of one surgeon were included in the follow-up study 2 to 15 years after operation. RESULTS: The 11 patients with polyposis all had "excellent" results. Of the 56 patients with ulcerative colitis, results were "good" or excellent in 48. Of the eight patients with less than good results, one died of complications of preexisting muscular dystrophy; four, or possibly six, had Crohn's disease. Presumably the original disease was Crohn's colitis instead of ulcerative colitis. CONCLUSIONS: The results of the study suggested that this operation was not appropriate for Crohn's disease and underscored the importance and the difficulties of differentiating the two conditions, if they are indeed separate entities. PMID- 8236011 TI - Exposure rates to patients' blood for surgical personnel. AB - BACKGROUND: Surgical personnel are at risk of contracting blood-borne diseases through exposure to patients' blood. Exposure rates for each surgical subspecialty have not been previously reported. The purpose of this study was to determine the rates of exposure to patients' blood for operating room personnel. METHODS: The study was conducted at Yale-New Haven Hospital, a level I trauma center and tertiary care hospital. During a 3-month period, exposed personnel were interviewed by a study nurse immediately after a cutaneous exposure to blood or after a sharp injury. RESULTS: During 2292 surgical procedures, 70 sharp injuries and 168 cutaneous exposures to blood were reported. The combined exposure rate (skin contact and sharp injury) was 10.4 per 100 procedures (95% confidence interval, 9.1 to 11.6) and ranged from 21.2 for general surgery to 3.3 for pediatric surgery (goodness-of-fit chi-squared, p < 0.001). The combined exposure rates were also significantly different among types of surgery and ranged from 18 for laparotomies to 4.3 for craniotomies (chi-squared, p < 0.001). The overall sharp injury rate was 3.1 per 100 procedures (95% confidence interval, 2.3 to 3.8) and ranged from 4.3 for general surgery to 1.3 for vascular surgery. CONCLUSIONS: The rate of exposure to blood for operating room personnel, which differ from prior studies, was 10.4 per 100 procedures and was highest for general surgical procedures. The differences in rates among studies might be attributable to different surgical technique, dissimilar case-mix, or different research methods relating to definition or ascertainment of exposure. PMID- 8236012 TI - Multifocal desmoids. AB - BACKGROUND: This study examines the natural history of patients treated for multifocal desmoids at a referral center for sarcomas. METHODS: Patients with multifocal desmoids seen at Memorial Sloan-Kettering Cancer Center in the 9-year period between 1982 and 1991 were identified through a prospective sarcoma data base. The clinical histories were reviewed. In addition, tumor estrogen and progesterone receptor status were evaluated by immunohistochemistry. RESULTS: This rare condition occurred in six (4.8%) of the 124 patients with desmoid tumors and all occurred in premenopausal women. In each instance the tumors were restricted to one anatomic region of the body and no patient had familial polyposis or Gardner's syndrome. When tested immunohistochemically, the tumors displayed no nuclear reactivity for estrogen or progesterone receptors. CONCLUSIONS: Further investigative work must be undertaken before considering routine use of antiestrogen therapy even in this subset of patients with desmoid tumors. PMID- 8236013 TI - Prevention of peritoneal adhesions by administration of sodium carboxymethyl cellulose and oral vitamin E. AB - BACKGROUND: Previous studies have shown that dietary supplements of vitamin E or the intraperitoneal administration of sodium carboxymethyl cellulose (SCMC) solution reduces postoperative adhesions by approximately 50%. The aim of this study was to determine whether there is a synergistic beneficial effect of vitamin E and SCMC in reducing postoperative adhesions. METHODS: Sixty Sprague Dawley rats were fed an identical diet containing 32 IU vitamin E/kg and were divided into four main groups: group A (control) and groups B, C, and D (experimental). Group D was further subdivided into three subgroups (D1, D2, and D3). Oral supplements of vitamin E in doses of 10 IU, 30 IU, and 30 IU/kg body weight were given to subgroups D1, D2, and D3 and group B, respectively, 5 days before operation and were continued until the fourteenth postoperative day when all animals were killed. Adhesions were created by scraping the cecum with mesh gauze followed by application of absolute alcohol and placement of silk suture in the parietal peritoneum. SCMC solution was administered intraperitoneally in groups C and D before closure. Adhesions were graded by two different investigators. RESULTS: All control animals developed significant adhesions, compared with no adhesions in 30% of group B (vitamin E) (p < 0.04), 40% in group C (SCMC) (p = 0.0001), and an average of 90% in the D groups (SCMC+vitamin E) (p = 0.0001). CONCLUSIONS: It appears that there is a synergistic beneficial effect of oral supplements of vitamin E and the intraperitoneal administration of SCMC solution in reducing the incidence and degree of intraperitoneal adhesions. PMID- 8236014 TI - Shear stress increases the release of interleukin-1 and interleukin-6 by aortic endothelial cells. AB - BACKGROUND: The aim of this study was to determine the correlation between shear stress and the release of interleukin-1 (IL-1) and interleukin-6 (IL-6) by endothelial cells (EC). METHODS: Bovine aortic EC were seeded in fibronectin coated cylinders at 1.0 x 10(6) cells/tube and allowed to reach confluence and to adhere for 48 hours. The experimental groups were subjected to a laminar flow of 100 ml/min (6 dyne/cm2). The control group was subjected to similar incubation conditions without flow. The release of IL-1 and IL-6 by EC was measured by enzyme-linked immunosorbent assay. RESULTS: Shear stress increased significantly (p < 0.01) the release of IL-1 and IL-6 by EC. The release of these two cytokines had different kinetics. CONCLUSIONS: Increasing shear stress facilitates release of IL-1 and IL-6 by EC. Previous reports have shown that IL-1 and IL-6 promote vascular smooth-muscle cell proliferation. Thus abnormal flow conditions with increasing shear stress may predispose to smooth-muscle cell proliferation that characterizes early atherosclerotic plaque development by an interleukin-mediated mechanism. PMID- 8236015 TI - Tyrosine inhibits intestinal-phase gastric-acid secretion. AB - BACKGROUND: Intestinal infusion of leucine augments the maximal gastric-acid secretory response to pentagastrin and serves as a bioassay for intestinal-phase acid secretion. Another action of leucine is to decrease tyrosine flux into neural tissues. We hypothesized that the mechanism of leucine-stimulated acid secretion involves its ability to alter tyrosine absorption. METHODS: We administered tyrosine intrajejunally and intravenously to anesthetized, vagotomized rats during maximal pentagastrin stimulation and measured acid output in response to intrajejunal infusion of leucine. RESULTS: Intrajejunal tyrosine produced a dose-dependent inhibition of leucine-stimulated acid secretion but only mild effects on acid secretion in response to pentagastrin alone. Intravenous tyrosine infusions also decreased acid secretion in response to intestinal leucine but required much higher doses. CONCLUSIONS: Tyrosine exerts a specific inhibitory effect on leucine-stimulated acid secretion, which is mediated intraluminally. Because tyrosine is an important regulator of adrenergic nervous system activity, we speculate that the mechanism of leucine-stimulated acid secretion may involve modulation of the sympathetic nervous system, thereby affecting acid secretion. PMID- 8236016 TI - Clinical significance of routine imaging of iliac and calf veins by color flow duplex scanning in patients suspected of having acute lower extremity deep venous thrombosis. AB - BACKGROUND: Because duplex ultrasonography is used increasingly to evaluate patients suspected of having acute deep venous thrombosis of the lower extremity, the clinical significance of limiting venous duplex scanning to the common femoral, superficial femoral, and popliteal veins becomes an important question. METHODS: We prospectively studied by venous color flow duplex ultrasonography 181 patients referred for evaluation to rule out acute deep venous thrombosis to determine (1) the frequency with which the iliac and calf veins could be imaged adequately to determine the presence or absence of acute deep venous thrombosis, (2) the frequency of acute venous thrombosis in these venous segments, and (3) the time required to scan these additional segments. RESULTS: In 79% of the patients studied, one segment of the iliac venous system was imaged adequately to determine the presence or absence of venous thrombosis. In the lower extremity, the common femoral vein, the superficial femoral vein, and the popliteal vein were imaged adequately in 94% of the patients. In 76% of the patients all three calf veins were imaged adequately to determine whether acute deep venous thrombosis was present or absent. The anterior tibial vein was the most difficult vein to image consistently and was imaged adequately in 76% of the patients referred. The mean time to scan the affected limb was 8 minutes 37 seconds. Forty seven patients (26%) of the 180 patients studied had positive venous scans. Twenty-three percent of the patients who had positive scans were found to have iliac vein involvement. Thirty-two percent who had positive venous scans were found to have thrombosis of the calf veins. Fifteen percent of the 47 patients who had positive scans had either an isolated iliac or calf vein thrombosis. CONCLUSIONS: We believe this frequency of isolated iliac or calf vein thrombosis coupled with the frequency of successful imaging of these venous segments and the short additional time required to image these segments justifies the inclusion of routine imaging of the iliac and calf veins in patients undergoing venous duplex scanning to rule out acute deep venous thrombosis. PMID- 8236017 TI - Technical considerations and complications associated with the placement of 180 implantable hepatic arterial infusion devices. AB - BACKGROUND: Treatment regimens with hepatic arterial chemotherapy infusion are being investigated in an attempt to improve survival and quality of life for patients with primary and metastatic liver malignancies. The successful delivery of chemotherapeutic drugs through an implantable hepatic arterial infusion device depends on the surgeon's understanding of hepatic arterial anatomy, the proper cannulation technique, and the operative measures necessary to prevent misperfusion of drug. METHODS: Between January 1, 1987, and December 31, 1991, we placed implantable hepatic arterial infusion devices in 180 patients. The records of these patients were review to determine (1) the incidence and surgical management of variant hepatic arterial anatomy and (2) the complications associated with surgical placement of these devices. RESULTS: Variant hepatic arterial anatomy requiring ligation of the variant vessel or nonstandard cannulation was seen in 66 patients (36.7%). Treatment response rates and duration of treatment were no different for these 66 patients than for the 114 patients with standard hepatic arterial anatomy (p = 0.94). There were no operative deaths in this series. Operative or early postoperative (within 30 days) complications occurred in 10 patients (5.5%). However, late complications or device-related malfunctions developed in 52 patients (28.8%). CONCLUSIONS: An understanding of regional arterial anatomy is required to surgically place a catheter to achieve bilobar hepatic arterial perfusion and avoid gastroduodenal misperfusion of drug. Placement of hepatic arterial infusion devices has a low rate of early morbidity, but surgeons should be aware of late complications that may develop in patients undergoing hepatic arterial chemotherapy infusion through an implantable device. PMID- 8236018 TI - Congenital hemangiopericytoma: an unusual vascular neoplasm of infancy. AB - BACKGROUND: Hemangiopericytoma is an uncommon tumor of infants, which originates from the vascular pericytes. Although generally considered to benign, metastases can occur. METHODS: Five cases of congenital hemangiopericytoma were seen in infants; all were found in females. The mean age at diagnosis was 7 weeks (range, birth to 10 weeks). The lesions were located in the neck, the parotid, the axilla, and the retroperitoneum. One neck lesion was detected prenatally by ultrasonography. Each lesion was resected. The diagnosis of congenital hemangiopericytoma was established only after histologic examination. RESULTS: No evidence of recurrence has been found in four of the children. However, intrathoracic and intracranial metastases developed in one child with a neck lesion 28 months after the original resection, and the child was treated with chemotherapy, but she died of progressive disease. CONCLUSIONS: We advocate the consideration of congenital hemangiopericytoma in the newborn infant with a vascular mass. Because congenital hemangiopericytoma is unresponsive to steroid therapy, unlike other vascular malformations, resection is the treatment of choice. Long-term postoperative follow-up is essential for the early detection of metastases. PMID- 8236019 TI - Characterization of cholecystokinin receptors on the human sphincter of Oddi. AB - BACKGROUND: The present in vitro study investigated the interaction between cholecystokinin (CCK) and receptors on human sphincter of Oddi tissue obtained from donated human livers that were being transplanted. METHODS: Radiolabeled ligands with cholecystokinin receptor specificity, autoradiography, and crystal scintillation counting were used to directly characterize cholecystokinin receptors on tissue sections. RESULTS: The binding of 125I-BH-CCK-8 to the tissue was saturable, specific, and dependent on time, pH, and temperature. Saturable binding of 125I-BH-CCK-8 was localized on the smooth muscle layer, and binding was inhibited only by cholecystokinin-related peptides. Computer analysis of 125I BH-CCK-8 binding indicated the presence of two classes of binding sites, one with a high affinity and the other with a low affinity for CCK-8. CCK-8 caused relaxation (half-maximal concentration, 6 nmol/L) and carbachol caused contraction (half-maximal concentration, 10 nmol/L) of circular, cross-sectional strips of the tissue. Longitudinal strips were less responsive. The relative 125I BH-CCK-8 binding inhibition potency of CCK-8 agreed closely with its relative ability to cause sphincter relaxation. Tetrodotoxin (1 mumol/L) and atropine (1 mumol/L) caused a rightward shift of the dose-response curve for CCK-8-stimulated sphincter relaxation. CONCLUSIONS: The present results indicate that cholecystokinin receptors on the human sphincter of Oddi are sulfate dependent and mediate sphincter relaxation. PMID- 8236020 TI - Time course and variability of the endocrine and metabolic response to severe sepsis. AB - BACKGROUND: In trauma patients a specific pattern of the hormone endocrine response to injury has been described. Conflicting data exist regarding the hormonal changes during sepsis. We studied the time course and variability of the hormonal and metabolic responses to severe sepsis in twelve medical patients. METHODS: Hormone levels were measured every 6 hours. Urinary nitrogen and 3 methylhistidine excretion were measured daily. Indirect calorimetry was used to calculate energy production rate (EPR). RESULTS: None of the hormones measured showed a time course in individual patients. Growth hormone (1.69 +/- 0.17 ng/ml) and insulin (48 +/- 5 mU/L) values were within normal range. Cortisol value (474 +/- 44 nmol/L) was in the upper normal range. Insulin-like growth factor 1 level (12.5 +/- 5.7 nmol/L) was decreased. Nitrogen balance was negative and did not change during the study period. Neither oxygen consumption (VO2) (318 +/- 20 ml/min), carbon dioxide output (283 +/- 10 ml/min), EPR (2064 +/- 435 kcal/day), nor fuel utilization showed a time course. The variability during the study period compared with values on admission ranged from 20% (insulin-like growth factor 1) to 74% (growth hormone). The variability of calculated fuel utilization ranged from 35% (protein) to 180% (fat). The variability of VO2, carbon dioxide production, and EPR was for less, 8%. CONCLUSIONS: A specific pattern in the endocrine and metabolic responses as observed in trauma patients does not occur in medical patients with severe sepsis. Hormones and calculated fuel utilization show large variations when compared with values on admission. VO2 and EPR vary for less. Nutritional needs in patients with sepsis, therefore, can be calculated on admission and need not be recalculated during the first week after admission. PMID- 8236021 TI - Nitric oxide-mediated, endothelium-dependent vasodilation is selectively attenuated in the postischemic extremity. AB - BACKGROUND: Attenuation of endothelium-dependent relaxing factor (EDRF) release may contribute to adverse sequelae commonly seen after reperfusion of an acutely ischemic extremity. The purpose of this study was to identify the compound responsible for the EDRF activity in the extremity and to evaluate its modulation by ischemia and reperfusion. METHODS: Isolated rat hindlimbs were perfused at constant pressure with an albumin-enriched crystalloid buffer. Increasing log dose infusions of acetylcholine and nitroprusside were used to measure endothelium-dependent (EDRF-mediated) and endothelium-independent vasoreactivity, respectively. RESULTS: Graded reductions in total vascular resistance were seen in response to both agonists in the control group (n = 11). In the postischemic group (n = 7), 60 minutes of normothermic ischemia and 10 minutes of reperfusion reduced endothelium-dependent vasodilation to acetylcholine by approximately 40% (p < 0.01). Endothelium-independent vasodilation to nitroprusside was unaffected. NG-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of nitric oxide release, attenuated vasodilation to acetylcholine (p < 0.01) but not nitroprusside in both control and postischemic groups. CONCLUSIONS: Endothelium dependent vasodilation in the rat hindlimb, mediated by nitric oxide, was selectively impaired by injury from ischemia and reperfusion. Strategies designed to minimize postischemic attenuation of nitric oxide release may be beneficial in the management of acute limb ischemia. PMID- 8236022 TI - Comparison of outcome between extended and nonextended liver resections for neoplasms. AB - BACKGROUND: An aggressive approach to liver resection for neoplasms was adopted during a 5-year period. The results were evaluated by comparing the outcome between extended and nonextended liver resections. METHODS: Among 73 consecutive patients undergoing liver resections for neoplasm or suspicion of neoplasm, 33 underwent extended liver resections (26 extended right lobectomies and 7 extended left lobectomies) and 40 underwent resections of lesser extent. Mortality, morbidity, and survival between the two groups were compared. RESULTS: Overall morbidity was 21% and perioperative mortality was 5.4%. Morbidity and mortality were not significantly different between extended and nonextended liver resections (24% vs 18% and 6.1% vs 5.0%, respectively). Liver resections for extrahepatic cholangiocarcinomas were found to be associated with a higher morbidity (6 of 10) and mortality (2 of 10) when compared with liver resections for noncholangiocarcinomas. Bile leaks occurred mainly in patients with cholangiocarcinomas (4 of 6) and contributed to an increased overall morbidity in this series. After a median follow-up of 31 months, there was no difference in survival between extended and nonextended liver resections. CONCLUSIONS: Extensive liver resections for neoplasms can be carried out without significant increase in the operative risk. Short-term survival is comparable between patients undergoing extended and nonextended liver resections. PMID- 8236023 TI - Hepatic vein reconstruction of the graft in partial liver transplantation from living donor: surgical procedures relating to their anatomic variations. AB - BACKGROUND: The surgical procedures to reconstruct the hepatic veins differ according to their anatomic variations to obtain the optimal graft volume for the recipient. This is an overview of the procedures used in our 25 living related liver transplantations. METHODS: The donor/recipient body weight ratio ranged widely from 1.2:1 to 9.6:1 (5.3 +/- 0.5:1, mean +/- SEM). RESULTS: The graft weight/recipient body weight was 2.40% +/- 0.21%. Graft components, which were determined by the optimal graft volume, and their drainage veins were the following: (1) segments 2 and 3 (S2+3) were used in 13 cases, 11 with the left hepatic vein (LHV) and two with the LHV and a partial drainage vein of S3; (2) S2+3 and a part of S4 in eight cases, seven with LHV and one with LHV and a partial drainage vein of S4; (3) S2+3+4 in three cases, with the common trunk of LHV and middle hepatic vein in all cases; and (4) S5+6+7+8 in one case with right hepatic vein. In two of three cases in which the graft had two drainage veins, the two vessels were reformed to have a common anastomotic orifice by the back table plastic surgery procedure. In the other case in which the procedure could not be performed, two separate anastomoses of the individual vessels were performed successfully. Although stenosis of the reconstructed hepatic veins occurred four times in two cases at 3 months or more after transplantation, all incidences could be completely repaired by balloon dilation. CONCLUSIONS: These results show that, with careful consideration of the hepatic vein reconstruction, pediatric patients can receive optimal volume grafts from living donors. PMID- 8236024 TI - Axial volvulus of the small bowel caused by Meckel's diverticulum. AB - BACKGROUND: A mechanism where the presence of a Meckel's diverticulum might cause bowel obstruction by twisting the small bowel along its long axis has been described. The developmental anatomy of the Meckel's diverticulum and its blood supply has been traced. Various other ways a Meckel's diverticulum can cause an obstruction have been enumerated. METHODS: We have reported two cases where the presence of a Meckel's diverticulum was the cause of bowel obstruction in a fashion hitherto not described. One patient had intermittent bouts of obstruction; the second patient had an acute obstruction. RESULTS: Preoperative investigations for a Meckel's diverticulum are not always fruitful, and most of the time the diagnosis is made at operation. When a Meckel's diverticulum is encountered at operation, division of its mesentery, the mesodiverticular band, rather than diverticulectomy, to relieve the volvulus described might devitalize the Meckel's diverticulum and should be avoided. CONCLUSIONS: A new mechanism by which the presence of a Meckel's diverticulum can cause an obstruction has been described. A short mesodiverticular band causes the bowel to twist along its long axis, precipitating the obstruction. This condition should be kept in mind in patients with a bowel obstruction who have not undergone any previous abdominal operation. PMID- 8236025 TI - Spontaneous bilateral pneumothorax--complication of laparoscopic cholecystectomy. AB - Pneumothorax is a rare complication of pneumoperitoneum. We report a case of spontaneous bilateral pneumothorax during laparoscopic cholecystectomy. The history, pathogenesis, and therapeutic options of this unusual occurrence are discussed. Because laparoscopic operations have become increasingly popular in recent years, we call attention to this rare but potentially life-threatening complication. PMID- 8236026 TI - When to perform either an axilloaxillary bypass graft or a carotid-subclavian bypass graft in patients with a symptomatic lesion of the subclavian artery. PMID- 8236027 TI - A thromboxane A2 synthetase inhibitor, OKY-046, reduces liver damage in hepatectomy. PMID- 8236028 TI - Association of annular pancreas and intestinal rotational defect in an adult. PMID- 8236029 TI - Actinobacillus actinomycetemcomitans and localized juvenile periodontitis. Clinical, microbiologic and histologic studies. AB - The present studies examined Actinobacillus actinomycetemcomitans and its role in localized juvenile periodontitis (LJP). The distribution of the bacteria was studied in healthy normals, patients with adult periodontitis, diabetics, and those with LJP. Over 95% of the LJP patients harbored A. actinomycetemcomitans, whereas only 17% of healthy subjects, 21% of adult periodontitis patients, and 5% of diabetics were positive. All members of a LJP family harboring the organism yielded isolates of the same biotype and serotype. The transmission of the bacteria was studied after transfer of the bacteria, with periodontal probes from infected to healthy gingival sites, within the oral cavity of LJP patients. Newly colonized gingival sites, 50% of those involved, became free of A. actinomycetemcomitans after only 3 weeks. A purposely forceful inoculation contributed to a more predictable colonization (89%), but only prolonged the colonization with one week. Treatment of LJP lesions with scaling and root planing resulted in minimal clinical and microbiological changes during a 16 week follow-up period. However, gingival curettage and modified Widman flap surgery suppressed A. actinomycetemcomitans in 75% and 89% of the sites, and resulted in resolution of periodontal pocket depth and gain in attachment level. Gingival tissue specimens, from 35 LJP sites, 3 control sites, and one monkey biopsy, were studied to verify the hypothesis of gingival infiltration of A. actinomycetemcomitans. Bacteria were identified immunohistologically with rabbit antisera serospecific to the three A. actinomycetemcomitans serotypes. Positive staining was observed in the tissue from all but one LJP patient. Twenty-eight (80%) lesions were positive for A. actinomycetemcomitans antigens in the gingival connective tissue, often with antigens located both between and within cells. The specimen from a culture positive control demonstrated no signs of invasion, similar to the monkey specimen. Transmission electron microscopic examination verified gram-negative bacteria that appeared as single microbes or small clusters in the connective tissue. The viability of the invading bacteria was studied by selective culture of minced tissue specimens and a series of washing solutions, after a rigorous surface disinfection. The washings successfully removed the bacteria in most cases, and after mincing 73% of the biopsies showed to be culture positive. The culture recovery of A. actinomycetemcomitans from minced tissues correlated exceedingly well with the presence of specific antigens in the gingival tissues from corresponding gingival biopsies. To further demonstrate the importance of A. actinomycetemcomitans in LJP, six culture positive LJP patients were treated by systemic tetracycline alone.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8236030 TI - Clinical and microbiological effects of subgingival application of a chlorhexidine gel in chronic periodontitis. A pilot study. AB - Clinical and microbiological effects of subgingival application of a 1% chlorhexidine gel were studied. Eleven patients with chronic periodontitis participated in the study. In each patient two pairs of single rooted teeth with at least 20% attachment loss and probing depth > 4 mm were selected. Contralateral sites were randomly allocated to test and control groups receiving chlorhexidine or placebo gel respectively. The gels were applied once daily for two consecutive days. The clinical parameters comprised plaque index, gingival index, probing pocket depth and bleeding on probing. The composition of the subgingival microbiota was estimated with use of dark field microscopy. Baseline measurements were taken two weeks prior to chlorhexidine application. Microbiological and clinical parameters were further monitored 2, 7, 14 and 28 days after the last gel application. Chlorhexidine gel application did not significantly change the composition of the microflora compared to placebo application. There were no major differences in plaque and gingival indices. Effects on probing pocket depth of the chlorhexidine gel application may have been disguised by an influence from the diameter of the needle with which the application was performed. Needles with a diameter of 25G resulted in increases in probing pocket depth which indicates a possible mechanical trauma. This effect was not observed when needles with a diameter of 23G were used. PMID- 8236031 TI - Comparative SEM studies of the enamel surface appearance following the use of glass ionomer cement and a diacrylate resin for bracket bonding. AB - The enamel surface appearance after the use of glass ionomer cement and a diacrylate resin for orthodontic bracket bonding was compared in an in vivo test using scanning electron microscopy and an enamel surface index system. Significant alterations in the enamel surface were found irrespective of the bonding procedure. The use of glass ionomer cement resulted however in a somewhat less affected enamel surface compared to the acrylate resin. PMID- 8236032 TI - Combined use of hydroxy-apatite and Tisseel in experimental bone defects in the rabbit. AB - An experimental study in the rabbit has been undertaken to evaluate differences in bone regeneration in and around solid (Alveograf) and porous (Interpore 200) hydroxyapatite granulae. Material was placed into experimentally made bone defects and in half of the defects HA was also mixed with fibrin sealant (Tisseel). Material alone or mixed with Tisseel was also placed subperiosteally adjacent to the mandibular bone. The observation time was six months. The study showed, however, no difference in bone regeneration around solid or porous hydroxyapatite granula nor could we see differences when mixed with Tisseel or not. Implant material placed subperiosteally did not induce bone formation nor did it provoke bone resorption. The use of fibrin sealant (Tisseel) made the hydroxyapatite material much more easy to handle and to retain in the tissue. PMID- 8236033 TI - A method for dental care activities time study using observer monitored counting of frequencies. AB - In dentistry, there is a need for better understanding of the distribution of the time spent on different activities at dental clinics. In order to investigate these resource aspects, both for the total time and for various categories of personnel and/or patients in dental clinics at the Public Dental Service (PD), Goteborg, Sweden, a time study method was used. The present study reports on the development of the PD's time study method (TSM) and analyses its functionality. The TSM, based on formulated criteria, used a method for an observer-monitored counting of frequencies. Specially trained observers registered the sampled observations for each professional, using a separate code list for each personnel group to which the professional belonged. The observer registered the on-going activity, not the duration of the procedure. Each observer followed several persons working in separated areas of the dental clinic. In 1989, the TSM was tested with the purpose of obtaining mean time values for dental care activities in the PD clinics performing general dental care. This study comprised all personnel categories in the clinic teams. Clinics were randomly selected and stratified according to the amount of dentistry for children in relation to the total time for dental care, reported as the parameter "dentist hours". Eight out of a total of 36 clinics were randomized, two from each strata.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236034 TI - The reporting of dental care. A comparison of time and activity related follow-up methods. AB - In 1989, a time study method was developed and tested by the Public Dental Service, Goteborg Sweden (PD). The purpose was to estimate the distribution of time used for different activities in the PD clinics supplying general dental care. Using the results from the test of the time study (TIA) the aim of the present study was to compare, evaluate and discuss the corresponding reported results from the administrative Electronic Data Processing (EDP) systems used for reporting dental care, and to analyze the parameters included as tools for reporting dental care procedures. This study has shown that values for diagnostic and preventive procedures were too high as reported, in relation to intervention procedures, whereas agreement was good with regard to the values in the intervention categories for dental procedures. The reported values from the EDP systems which are based on the codes in the Swedish dental insurance tariff, represent dental care in the intervention categories. However, one must take account of the verified divergences in reporting values concerning all three main areas of dental procedures. Correct time reporting for the main sectors of dental procedures--diagnostic, prevention and intervention sectors--will improve the validity of EDP systems for cost report. PMID- 8236035 TI - Your money or your life. What health care reformists don't tell you as the hour of change draws near. PMID- 8236036 TI - The 1992 C.T. Rowland Award orthodontic case reports. PMID- 8236037 TI - Dentist volunteers needed in third world countries. PMID- 8236038 TI - Russian stomatology in the nineties: a Texas dentist observes it first-hand. PMID- 8236039 TI - Women in medicine not just a newfangled idea. PMID- 8236040 TI - Women's health is focus of continuing education course. PMID- 8236041 TI - Permanently unconscious patients: a radical proposal. PMID- 8236042 TI - TMA launches drive for border health commission. PMID- 8236043 TI - Tibet to Austin: physician meets medical needs of refugees. PMID- 8236044 TI - Houston scientists to track Chernobyl victims. PMID- 8236045 TI - Prevention remains center of attention at international AIDS conference. PMID- 8236046 TI - TDH helps physicians meet HIV-infected patients' needs. PMID- 8236047 TI - Managed care Q&A. PMID- 8236048 TI - All the right moves. Playing by new rules, medicine planned early and won big in the 73rd Texas Legislature. PMID- 8236049 TI - Texas researchers announce two major genetics breakthroughs. PMID- 8236050 TI - TMA studies options after state freezes Medicaid payments. PMID- 8236051 TI - Controversy over the silicone gel breast implant: current status and clinical implications. AB - The silicone gel breast implant controversy has generated much confusion, making informed decision making for physicians and patients increasingly difficult. In order to better understand the controversy, we review the steps taken by the Food and Drug Administration and the resultant status of breast implants. The currently available scientific data are summarized with regard to cancer development and detection, implant rupture and gel migration, and autoimmune disorders. By reviewing the presently available information on silicone gel breast implants, we can better address the concerns of our patients. PMID- 8236053 TI - HMO tactics hurt solo specialist's practice. PMID- 8236052 TI - Subtle subluxation of Lisfranc's joint. PMID- 8236054 TI - Lawsuit abuse continues to plague Valley physicians. PMID- 8236055 TI - Review of occupational health study clarified. PMID- 8236056 TI - First woman, first RN, chairs TDH Board of Health. An exclusive interview with Ruth Stewart. PMID- 8236057 TI - Reflections of a Vietnam nurse. PMID- 8236058 TI - ANA president answers nurses questions. PMID- 8236059 TI - Self-organization: the basic principle of neural functions. AB - Recent neurophysiological observations are giving rise to the expectation that in the near future genuine biological experiments may contribute more than will premature speculations to the understanding of global and cognitive functions. The classical reflex principle--as the basis of neural functions--has to yield to new ideas, like autopoiesis and/or self-organization, as the basic paradigm in the framework of which the essence of the neural can be better understood. Neural activity starts in the very earliest stages of development well before receptors and afferent input become functional. Under suitable conditions, both in nervous tissue cultures and in embryonic tissue recombination experiments, the conditions of such initial autopoietic activity can be studied. This paper tries to generalize this elementary concept for various neural centers, notably for the spinal segmental apparatus and the cerebral cortex. PMID- 8236060 TI - The visual system and levels of perception: properties of neuromental organization. AB - To see whether the mental and the neural have common attributes that could resolve some of the traditional dichotomies, we review neuroscientific data on the visual system. The results show that neuronal and perceptual function share a parallel and hierarchical architecture which is manifest not only in the anatomy and physiology of the visual system, but also in normal perception and in the deficits caused by lesions in different parts of the system. Based on the description of parallel hierarchical levels of active information processing in the visual brain, we suggest a concept of dissociable levels of perception, advocating that the phenomenal perception and recognition is realized in the functional integrity of a network of reciprocal cortico-cortical connections. The properties shared by neuronal and perceptional functions provide a basis for a neuromental monism in which both functions are attributed a causal role. PMID- 8236061 TI - Neurodynamic system theory: scope and limits. AB - This paper proposes that neurodynamic system theory may be used to connect structural and functional aspects of neural organization. The paper claims that generalized causal dynamic models are proper tools for describing the self organizing mechanism of the nervous system. In particular, it is pointed out that ontogeny, development, normal performance, learning, and plasticity, can be treated by coherent concepts and formalism. Taking into account the self referential character of the brain, autopoiesis, endophysics and hermeneutics are offered as elements of a poststructuralist brain (-mind-computer) theory. PMID- 8236062 TI - A discussion of the mind-brain problem. AB - In this paper Popper formulates and discusses a new aspect of the theory of mind. This theory is partly based on his earlier developed interactionistic theory. It takes as its point of departure the observation that mind and physical forces have several properties in common, at least the following six: both are (i) located, (ii) unextended, (iii) incorporeal, (iv) capable of acting on bodies, (v) dependent upon body, (vi) capable of being influenced by bodies. Other properties such as intensity and extension in time may be added. It is argued that a fuller understanding of the nature of forces is essential for the analysis of the mind-brain problem. The relative autonomy and indeterministic nature of mind is stressed. Indeterminism is treated in relation to a theorem of Hadamard. The computer theory of mind and the Turing test are criticized. Finally the evolution of mind is discussed. PMID- 8236063 TI - Mental illness and the mind-brain problem: delusion, belief and Searle's theory of intentionality. AB - Until recently there has been little contact between the mind-brain debate in philosophy and the debate in psychiatry about the nature of mental illness. In this paper some of the analogies and disanalogies between the two debates are explored. It is noted in particular that the emphasis in modern philosophy of mind on the importance of the concept of action has been matched by a recent shift in the debate about mental illness from analyses of disease in terms of failure of functioning to analyses of illness in terms of failure of action. The concept of action thus provides a natural conduit for two-way exchanges of ideas between philosophy and psychiatry. The potential fruitfulness of such exchanges is illustrated with an outline of the mutual heuristic significance of psychiatric work on delusions and philosophical accounts of Intentionality. PMID- 8236064 TI - Neuroscience and the problem of consciousness: theoretical and empirical approaches. An introduction. PMID- 8236065 TI - Drawing the boundary between subject and object: comments on the mind-brain problem. AB - Physics says that it cannot deal with the mind-brain problem, because it does not deal in subjectivities, and mind is subjective. However, biologists (among others) still claim to seek a material basis for subjective mental processes, which would thereby render them objective. Something is clearly wrong here. I claim that what is wrong is the adoption of too narrow a view of what constitutes 'objectivity', especially in identifying it with what a 'machine' can do. I approach the problem in the light of two cognate circumstances: (a) the 'measurement problem' in quantum physics, and (b) the objectivity of standard mathematics, even though most of it is beyond the reach of 'machines'. I argue that the only resolution to such problems is in the recognition that closed loops of causation are 'objective'; i.e. legitimate objects of scientific scrutiny. These are explicitly forbidden in any machine or mechanism. A material system which contains such loops is called 'complex'. Such complex systems thus must possess non-simulable models; i.e. models which contain impredicativities or 'self-references' which cannot be removed, or faithfully mapped into a single coherent syntactic time-frame. I consider a few of the consequences of the above, in the context of thus redrawing the boundary between subject and object. PMID- 8236066 TI - European standards for lung function testing: 1993 update. PMID- 8236067 TI - Effect of air pollution on the prevalence of asthma and allergy: lessons from the German reunification. PMID- 8236068 TI - Acute effects of inhaled salbutamol on the metabolic rate of normal subjects. AB - BACKGROUND: This study was designed to investigate the contribution of inhaled salbutamol to the increase in resting metabolic rate found in patients with chronic airflow limitation who were receiving bronchodilator therapy. METHODS: The resting metabolic rate of 10 normal subjects (age 20-47 years, weight 42-105 kg, seven men) was studied after inhalations of salbutamol or placebo. An open canopy method of indirect calorimetry was used to measure resting oxygen consumption (VO2) and resting carbon dioxide production (VCO2). Subjects inhaled two, four, eight, or 12 puffs (100 micrograms/puff) of salbutamol or placebo in a double blind manner. Recordings of VO2 and VCO2 were made after inhalation of the four doses of salbutamol or placebo, integrated over one hour, and compared. RESULTS: VO2 and VCO2 increased in a dose dependent manner after inhaled salbutamol with a maximum effect at five minutes after inhalation. After four puffs, VO2 was 203 and 188 ml/kg/h for salbutamol and placebo respectively. After eight puffs, VO2 was 207 and 185 and VCO2 was 167 and 155 ml/kg/h. After 12 puffs, VO2 was 220 and 190 with a VCO2 of 181 and 168 ml/kg/h. Twelve puffs of salbutamol increased the mean (SE) respiratory quotient from 0.85 (0.01) to 0.93 (0.04) at five minutes indicating an increase in ventilation in excess of metabolic demand. Mean heart rate increased in parallel with VO2. CONCLUSION: Inhaled salbutamol significantly increases resting metabolic rate in a dose dependent manner. PMID- 8236069 TI - Modification of the thermogenic effect of acutely inhaled salbutamol by chronic inhalation in normal subjects. AB - BACKGROUND: Acute inhalation of clinical doses of salbutamol in normal volunteers increases resting metabolic rate by up to 20% above control values. This study was designed to see if chronic treatment with salbutamol causes a sustained increase in metabolic rate and whether it modifies the acute thermogenic response to the drug. METHODS: The effects of chronic inhaled salbutamol on resting oxygen consumption (VO2) and carbon dioxide output (VCO2) were studied in seven normal subjects (age 20-47 years, weight 52-105 kg, five men). An open canopy method of indirect calorimetry was used to measure VO2, VCO2, and respiratory quotient (RQ). Subjects inhaled two puffs of salbutamol or placebo four times a day in a double blind manner. Measurements of resting VO2 and VCO2 after 10 days of salbutamol were compared with the values after 10 days of placebo and with those taken at the start of the study. At the end of each treatment period subjects inhaled eight puffs (800 micrograms) of salbutamol and the acute effects on VO2, VCO2 and RQ were monitored for one hour. RESULTS: Resting VO2, VCO2, and RQ were not significantly different at the end of the salbutamol and placebo periods but the acute response to eight puffs of salbutamol was abolished by regular inhalation. The mean VO2 integrated over one hour after 800 micrograms salbutamol given acutely was different (241.3 and 210.7 ml/kg/h in the placebo and salbutamol groups respectively). Differences were not significant between placebo and salbutamol groups for changes in VCO2, heart rate, blood pressure, and RQ after acute inhalation. CONCLUSION: Regular treatment with inhaled salbutamol (800 micrograms/day) does not cause a sustained increase in resting metabolic rate but prevents the increase in VO2 that occurs after acute inhalations in normal subjects. PMID- 8236070 TI - Effect of orally administered beclomethasone dipropionate on calcium absorption from the gut in normal subjects. AB - BACKGROUND: There is evidence that patients with chronic obstructive airways disease and asthma who take inhaled steroids have a low bone density. As most of a drug given from a metered dose inhaler is actually swallowed, the possibility that swallowed beclomethasone dipropionate acts topically in the gut to impair calcium absorption was investigated. Such an effect, if sustained, may be a causative factor of long term bone loss. METHODS: A two week randomised, double blind, placebo controlled, crossover trial was performed in 12 normal volunteers. Subjects were randomly allocated to swallow beclomethasone dipropionate capsules (500 micrograms twice a day) or placebo for one week. The alternate capsule was given throughout the second week. At the end of each week, calcium absorption was assessed by a strontium absorption test. Serum parathyroid hormone, plasma calcium, and plasma phosphate concentrations were determined on the last two days of each week. Twenty four hour urinary calcium, hydroxyproline, and cortisol concentrations were measured for four successive days in each week. RESULTS: All subjects completed the study. There was a 12% reduction in strontium absorption during the beclomethasone dipropionate ingestion week. There was also a 23% reduction in 24 hour urinary cortisol excretion during the same week. CONCLUSIONS: Calcium absorption (measured by a strontium absorption test) was reduced by oral administration of beclomethasone dipropionate for one week. Decreased calcium absorption due to swallowed corticosteroid may contribute to side effects of inhaled steroids and further long term studies are needed. PMID- 8236071 TI - Reproducibility of incremental maximal cycle ergometer testing in patients with restrictive lung disease. AB - BACKGROUND: Exercise testing has become an important tool in the diagnosis and treatment of restrictive lung disease. The reproducibility of variables measured during exercise testing was examined in subjects with stable restrictive lung disease. METHODS: Six subjects, who had never previously undergone exercise testing, each underwent three maximal incremental exercise studies on a bicycle ergometer conducted during a 28 day period. RESULTS: Data collected at rest, before exercise, were not significantly different during the three study days. Comparison of results at the end of the exercise tests from the three studies also revealed no evidence of a significant learning effect. Reproducibility of exercise performance by subjects was assessed by the coefficient of variation. The mean within subject coefficient of variation at the end of the exercise tests was 5.6% for work rate, 7.9% for exercise duration, and 9.5% for dyspnoea. The mean within subject coefficient of variation obtained at the end of the exercise tests was 5.3% for oxygen uptake (VO2), 2.5% for oxygen saturation (SaO2), 4.0% for heart rate (HR), 5.5% for minute ventilation (VE), 5.8% for respiratory frequency (f), and 4.6% for tidal volume (VT). The mean within subject coefficient of variation at 40% and 70% of maximal work rates for VO2 was 5.7% and 5.6% respectively, for SaO2 1.3% and 1.5%, for HR 4.8% and 4.0%, for VE 6.3% and 6.6%, for f 10.1% and 7.8%, and for VT 6.0% and 4.5%. CONCLUSIONS: Variables measured during clinical exercise testing in subjects with restrictive lung disease are highly reproducible. No significant learning effect was found on repeated testing in subjects who had never previously undergone exercise testing. PMID- 8236072 TI - Factors affecting peak expiratory flow variability and bronchial reactivity in a random population sample. AB - BACKGROUND: Bronchial reactivity measurements are widely used in epidemiological studies to provide an objective marker of asthma. There are, however, several potential advantages of measuring peak expiratory flow (PEF) variability instead, particularly in large studies. PEF variability and bronchial reactivity were compared in a population sample to assess the relationships of the two measurements to factors known to be associated with airways disease, and to compare their response rates. METHODS: Subjects aged 18-65 were randomly selected from the electoral register of an administrative area in eastern England and randomised to attend either for a bronchial challenge test measuring the provocative dose of methacholine producing a 20% fall in FEV1 (PD20), or to measure PEF at two hourly intervals during waking hours for one week. Skin tests with common allergens were performed and a smoking history obtained. PEF variability was expressed as the amplitude % mean (highest - lowest x 100/mean). RESULTS: A total of 273 subjects (69%) collected a PEF meter but a completed record sheet was returned by only 247 (62%); this was still significantly more than the 202 subjects (54%) who attended for and successfully completed a challenge test. Amplitude % mean was higher in women than in men (9.7% v 8.5%). In multiple regression analysis amplitude % mean increased significantly with age, mean skin weal diameter, and with current smoking. The odds of having a PD20 below 24.5 mumol increased with mean skin weal diameter and were greater in current smokers. Neither age nor sex had a significant effect on bronchial reactivity but there were significant interactions between age and the effects of both smoking and atopy. CONCLUSIONS: The higher response rate associated with the use of PEF variability measurement, and the association with factors implicated in the pathogenesis of airways disease, suggest that PEF variability would be a useful measurement to employ in epidemiological studies. PMID- 8236073 TI - Ethnic differences in lung function in Israeli children. AB - BACKGROUND: The population of Israel consists of immigrants from many different countries. It is not known whether a single nomogram can be used for spirometric values of children of different ethnic descent. METHODS: Spirometry was performed in 753 second or third generation Israeli children (7-14 years) of different ethnic groups. Both parents of 503 of the children were of the same ethnic background. Subjects were allocated to six ethnic groups (European, Iraqi, North African, Indian, Yemenite, and Georgian). RESULTS: Standing height contributed most to the prediction of spirometric values (forced expiratory volume in one second, forced vital capacity), whereas sitting height did not contribute further. Statistical analysis showed significant ethnic differences. The Georgians had higher spirometric values for FEV1 than all the other ethnic groups, and higher FVC values than those of the Yemenite, North African, and Indian groups. FVC was lower among the Indian than all other groups. CONCLUSION: Differences in normal spirometric values were found among second or third generation Israeli children of different ethnic origins. European, North African, Iraqi, and Yemenite children could be characterised by single equation, whereas children of Georgian and Indian descent needed different predicting equations. PMID- 8236074 TI - Daytime sleepiness, cognitive performance and mood after continuous positive airway pressure for the sleep apnoea/hypopnoea syndrome. AB - BACKGROUND: Patients with the sleep apnoea/hypopnoea syndrome often receive continuous positive airway pressure to improve their symptoms and daytime performance, yet objective evidence of the effect of this treatment on cognitive performance is lacking. METHODS: A prospective parallel group study was performed comparing the change in objective daytime sleepiness as assessed by multiple sleep latency, cognitive function, and mood in 21 patients (mean (SE) number of apnoeas and hypopnoeas/hour 57 (6)) who received continuous positive airway pressure for three months and 16 patients (49(6) apnoeas and hypopnoeas/hour) who received conservative treatment for a similar period. RESULTS: Both groups showed significant within group changes in cognitive function between baseline and three months, but when comparisons were made between groups the only significant difference was a greater improvement in multiple sleep latency with continuous positive airway pressure. However, the improvement in sleep latency with continuous positive airway pressure was relatively small (3.5 (0.5) to 5.6 (0.7) min). The group treated with continuous positive airway pressure was divided into those who complied well with treatment (> 4.5 hours/night) and those who did not. Those who complied well (n = 14) showed significant improvement in mean sleep latency and also in depression score compared with the controls but no greater improvement in cognitive function. CONCLUSION: This study confirms significant improvements in objective sleepiness and mood with continuous positive airway pressure, but shows no evidence of major improvements in cognitive function. PMID- 8236075 TI - Thoracoscopy assisted pulmonary lobectomy. AB - BACKGROUND: This report describes a preliminary experience with six patients undergoing video imaged thoracoscopic pulmonary lobectomy. METHODS: Three left upper lobectomies, and one each of right upper, right lower and left lower lobectomy were undertaken. The resections were performed as orthodox dissectional lobectomy procedures but were carried out under videothoracoscopic imaging with instruments introduced through two stab incisions. The entire resected lobe was delivered through a 7 cm submammary intercostal incision. RESULTS: There were no operative deaths or complications attributable to the technique. In three other patients conversion to an open thoracotomy was required because of bleeding (two cases) or obscure anatomy (one case). Post-operative pain in those undergoing thoracoscopic resection was less than that encountered with standard thoracotomy and early clinic review showed the patients to be pain free with excellent shoulder movement. CONCLUSIONS: Major pulmonary resection according to standard cancer practices is feasible with videothoracoscopic techniques. This approach is likely to offer considerable functional benefit to patients. Specimen delivery through the submammary incision imposes a 5 cm primary lesion size limitation. Detailed mediastinal assessment is necessary to exclude N2 status before undertaking thoracoscopic surgery. PMID- 8236076 TI - Empyema thoracis: a problem with late referral? AB - BACKGROUND: Patients are often referred to thoracic units for management of empyema after the acute phase has been treated with antibiotics but without adequate drainage. This study evaluates the effects of delay in surgical treatment of empyema thoracis on morbidity and mortality. METHODS: Thirty nine consecutive patients were studied from January 1991 to June 1992. Two groups (group 1, 16 patients; group 2, 23 patients) were compared depending on the time spent under the care of other specialists before referral to the thoracic unit (group 1, seven days or less; group 2, eight days or more). The reasons for delay in referral were analysed. RESULTS: Four patients were treated conservatively with chest drainage alone (all in group 1). Thirty five patients required rib resection and drainage of their empyema (group 1, 12 patients; group 2, 23 patients). Nineteen (all in group 2) of the 35 patients who had rib resections went on to have decortication. The commonest cause of empyema was post-pneumonic (37 out of 39 patients). Staphylococcus aureus was the commonest organism isolated. Misdiagnosis (five patients), inappropriate antibiotics (six patients), and inappropriate placement of chest drainage tubes (three patients) all contributed to persistence and eventual progression of empyema. The overall mortality was 10% and mortality increased with age. The median stay in hospital was 9.5 days (range 7-12 days, n = 4) for patients treated with closed tube drainage only; 18 days (range 10-33 days, n = 16) for patients who had undergone rib resections and open drainage; and 28 days (range 22-49 days, n = 19) for patients who underwent decortication. The likelihood of having a staged procedure (antibiotics, closed tube drainage, open drainage with rib resection, and finally decortication) increased when closed tube drainage was persevered with for more than seven days. The total hospital stay was positively related with the time before referral for surgical treatment. Anaemia, low albumin concentrations, and worsening liver function were found in group 2 compared with group 1. CONCLUSIONS: Early adequate operative drainage in patients with empyema results in low morbidity, shorter stays in hospital, and good long term outcome. These patients should be treated aggressively and early referral for definitive surgical management is recommended. PMID- 8236077 TI - Treatment of symptomatic pulmonary aspergillomas with intracavitary instillation of amphotericin B through an indwelling catheter. AB - BACKGROUND: The treatment of symptomatic pulmonary aspergillomas can be difficult. One approach has been to deliver antifungal drugs directly into the lung cavity. The use of this method of treatment is described in which an indwelling percutaneous catheter is used which avoids repeated needlings of the cavity and may allow extended treatment on a domiciliary basis. METHODS: Amphotericin B was delivered through indwelling percutaneous intracavitary catheters to treat five symptomatic episodes in four patients with pulmonary aspergillomas. RESULTS: The treatment was well tolerated by all patients and their symptoms resolved in four of the five episodes. Two patients died, one from an unrelated bronchopneumonia and the other from advanced cachexia. Two patients remain symptom free after eight and 12 months. CONCLUSIONS: Intracavitary administration of amphotericin through an indwelling catheter should be considered for any patient who has troublesome sputum production, haemoptysis, or systemic symptoms attributable to an aspergilloma. PMID- 8236078 TI - Cytokines and pulmonary fibrosis. AB - Chronically inflamed and fibrotic tissue of the respiratory tract can be shown to actively express the genes and products of a number of powerful growth and differentiating factors. The initial activation of lung inflammatory cells, including alveolar macrophages, is presumed to result in the release of early acting cytokines such as IL-1 and TNF. Subsequent activation and possible phenotype alteration of the structural cells results in release of other growth factors and accumulation of blood derived inflammatory cells. These cells, once they have entered the tissue and become further activated, may begin to release their own autocrine factors and "feed back" some of the similar signals to the tissue cells in a paracrine manner, further inducing differentiation and phenotype change. These internal tissue cell and cytokine cascades could account for the chronic nature of the inflammation. Therapeutic intervention must therefore take into account the inflammatory component as well as the nature of the cytokines and structural cells involved in the propagation of the disease. PMID- 8236080 TI - Successful treatment of diffuse lipoid pneumonitis with whole lung lavage. AB - Diffuse lipoid pneumonitis is rare. Prednisolone can be beneficial, but no other method of treatment has been tried. The first case of diffuse lipoid pneumonitis successfully managed with whole lung lavage is described. PMID- 8236079 TI - Exercise in patients with chronic obstructive pulmonary disease. AB - Sporadic visits to the local doctor followed sometimes by changes in oral and inhaled bronchodilators and occasionally by the addition of steroids frequently does little to significantly improve symptoms and function in the disabled patient with COPD. As in other chronic diseases, the management of these patients is facilitated by a team approach in conjunction with general rehabilitation principles. The rationale and practical implementation of such a programme has recently been outlined by the American Association of Cardiopulmonary Rehabilitation. These are multifaceted programmes but a key component, as outlined above, is exercise training. In this brief review the various approaches available have been described. Controversy still reigns regarding the optimal modes of training and there are important differences among the several approaches. Two main groups can be delineated. One emphasises the detailed definition of the impaired physiology with therapeutic measures targeted to specific defects. There is good documentation that, conversely, unstructured programmes that use treadmill and free range walking and cycling also improve endurance for walking. Upper extremity training is of additional benefit. Programmes with as little as three sessions per week of 1-2 hours of low intensity activity have achieved success so we know that simple programmes can be helpful. Moreover, without the necessity for complex testing and training methods these programmes can be implemented with relatively low costs. Future investigations to examine the relationship between improved exercise capacity for walking and arm exercise on the one hand, and the ease of performance of activities of daily living on the other, will help to reinforce the effectiveness of exercise programmes. PMID- 8236081 TI - Amyloidosis complicating cystic fibrosis. AB - Two patients with cystic fibrosis developed acute onset nephrotic syndrome and died within three months of presentation. Examination of renal biopsy specimens indicated amyloid. The onset of proteinuria or a fall in baseline renal function should alert the physician to this rare complication. PMID- 8236082 TI - Unusual presentation of a large tension bronchogenic cyst in an adult. AB - A routine chest radiograph in a 20 year old man revealed a giant air filled cavity of the left hemithorax under tension. At thoracotomy a large left lower lobe intraparenchymal cyst required lobectomy and the pathological findings were consistent with a bronchogenic cyst. Although tension bronchogenic cysts are common in infants, this case demonstrates their unusual presentation in an adult. PMID- 8236083 TI - Molecular biology of hemophilia B. PMID- 8236084 TI - Vascular smooth muscle cell proliferation: basic investigations and new therapeutic approaches. AB - Our investigations are based upon the hypothesis that synthesis and release of numerous mitogens allow interaction with a correspondingly large set of specific receptors to initiate a variety of secondary messages which eventually lead to SMC proliferation and migration. Unfortunately, the experimental tools available for studying these critical biologic systems under in vivo conditions are quite limited. Previous investigators have partially elucidated the growth factors responsible for intimal accumulation by infusing specific antibodies against individual mitogens such as PDGF or bFGF. However, no general approach was available to identify critical intracellular intermediates of the above pathway. We thought that antisense oligonucleotides might be useful in pinpointing these key components by specifically inhibiting their expression and then determining the effects of the suppression on SMC response in vivo. Data summarized above strongly imply that c-myb is a critical intermediate of a common mitogenic pathway required by all growth factors involved in SMC migration and growth. Given the previously documented in vitro effect of antisense oligonucleotide, we believe that the proto-oncogene is involved in proliferation. The drug heparin and blood vessel wall heparan sulfates probably also inhibit SMC growth via suppression of c-myb. Thus, endothelial cell and SMC heparan sulfates appear to constitute a natural blood vessel wall antiproliferative pathway. To the best of our knowledge, these studies constitute the first reported use of antisense oligonucleotide to inhibit synthesis of a normal gene product under in vivo conditions with a subsequent effect upon a cellular process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236085 TI - von Willebrand factor and cardiovascular disease. PMID- 8236086 TI - Mechanisms of shear-induced platelet adhesion and aggregation. PMID- 8236087 TI - Endothelium as a signal transduction interface for flow forces: cell surface dynamics. PMID- 8236088 TI - The effects of flow on blood coagulation and thrombosis. PMID- 8236089 TI - Regulation of PAI-1 gene expression in vivo. PMID- 8236090 TI - Plasminogen activator inhibitor 1 and atherothrombosis. PMID- 8236091 TI - Biology and facultative secretion of plasminogen activator inhibitor-2. PMID- 8236092 TI - Endothelial-leukocyte adhesion molecules in inflammation and metastasis. AB - Vascular endothelium is an active tissue. In response to various mediators, it expresses specific cell surface glycoproteins that support the binding and extravasation of blood leukocytes. These events are essential for host defense against pathogens and for repair of tissue damage. Unfortunately, it appears that a number of blood borne tumor cells may be able to use the same adhesive structures to facilitate their metastasis. The development of agents designed to block the adhesion of leukocytes (or tumor cells) to the vessel wall may prove valuable in the treatment of a variety of human disease processes, including adult respiratory distress syndrome (ARDS), ischemic reperfusion injury, autoimmunity, and metastatic spread of cancer. PMID- 8236093 TI - The role of thrombin and thrombin receptor activating peptide (TRAP-508) in initiation of tissue repair. PMID- 8236094 TI - Signaling pathways of the thrombin receptor. PMID- 8236095 TI - Protease nexin-1 and thrombin: injury-related processes in the brain. PMID- 8236096 TI - Modern treatment of hemophilia: from the shadows towards the light. PMID- 8236097 TI - Neointimal proliferation: the role of basic FGF on vascular smooth muscle cell proliferation. PMID- 8236098 TI - TGF-beta: structure, function, and formation. PMID- 8236099 TI - Gene induction in vessel wall injury. AB - VSMC have been long thought to play a critical role in restenosis by proliferating and migrating from the vessel media to the intima, resulting in intimal hyperplasia. Recently, it has been suggested that the VSMC may play an earlier role in the events leading to restenosis, for example through the production of PDGF. The work described above suggests that the VSMC may mediate both the early inflammatory and thrombotic responses associated with vessel injury. Thus the VSMC may be involved in all phases of vascular injury, including thrombosis, inflammation, and intimal hyperplasia. Additional work will be necessary to fully elucidate the programs activated in VSMC in response to growth and migratory factors. The recent advances in recombinant DNA technology provide the hope that this will lead to novel approaches to attenuate the response of the VSMC to injury. PMID- 8236100 TI - Thrombin receptor structure and function. PMID- 8236101 TI - The vascular angiotensin (AT1) receptor. PMID- 8236102 TI - Gene therapy of hemophilia B. PMID- 8236103 TI - Gene transfer in the treatment of thrombosis. PMID- 8236104 TI - Direct gene transfer: basic studies and human therapies. PMID- 8236105 TI - Low molecular weight heparin. PMID- 8236106 TI - Thrombin, thrombin inhibitors, and the arterial thrombotic process. PMID- 8236107 TI - Structural and functional characterization of tick anticoagulant peptide (TAP): a potent and selective inhibitor of blood coagulation factor Xa. AB - The discovery of rTAP has provided a wealth of important scientific information ranging from the structural and kinetic characterization of a novel serine protease inhibitor to the importance of factor Xa in the thrombotic process. The results obtained with this inhibitor thus far have broadened our understanding of hemostasis and thrombosis and ultimately may result in the development of newer and more efficient therapeutic agents to combat one of the leading causes of morbidity and mortality in man today. PMID- 8236108 TI - Signaling through G proteins and G protein-coupled receptors during platelet activation. AB - Recent studies have helped to define the early events of signal transduction in platelets. The best-described of these events are those in which heterotrimeric guanine nucleotide binding regulatory proteins (G proteins) mediate the interaction between cell surface receptors for agonists and intracellular second messenger generating enzymes. To date nine different G proteins have been identified in platelets. Their targets include phospholipases C and A2, and adenylyl cyclase. Efforts to clone the receptors that can couple to these G proteins have been successful for epinephrine, thrombin, TxA2 and platelet activating factor. Each of these is comprised of a single polypeptide with seven transmembrane domains and an extracellular N-terminus. In the case of the thrombin receptor, activation occurs by a novel mechanism in which thrombin cleaves its receptor, creating a new N-terminus that can serve as a tethered ligand. Shortly after activation, thrombin receptors become resistant to re activation by thrombin. This desensitization, which appears to involve receptor phosphorylation and internalization, provides a potent mechanism for limiting the duration of thrombin-initiated events in platelets and other thrombin-responsive vascular cells. PMID- 8236109 TI - Mechanisms of cytoskeletal reorganization during platelet activation. PMID- 8236110 TI - The evolution of vertebrate blood coagulation: a case of Yin and Yang. PMID- 8236111 TI - Molecular events that control the protein C anticoagulant pathway. AB - Despite its rather recent identification, the protein C activation system has afforded many investigators with unique opportunities to probe the molecular basis by which cofactors function. Thrombomodulin clearly exerts its specificity switch both by interacting directly with the fibrinogen binding site on thrombin (exosite 1) and by altering the conformation within the enzyme center. At least in the case of thrombomodulin, these conformational changes appear to overcome repulsive interactions between acidic residues in the substrate and the enzyme. To determine whether the models derived from attempts at the molecular analysis of the protein C activation complex are at all relevant to the other coagulation complexes will require further examination, but the concept that residues near the cleavage site contact residues in the free enzyme in an unfavorable fashion, and that the cofactors overcome these inhibitory interactions is a hypothesis that is directly testable for all of the complexes. The availability of crystal structures for the coagulation enzymes, coupled with the capacity to mutagenize both the substrate and the enzyme, promises to provide new insights into molecular events that control coagulation. PMID- 8236112 TI - Human fibrinogen--structural variants and functional sites. PMID- 8236113 TI - Primitive coagulation systems and their message to modern biology. PMID- 8236114 TI - Treatment of patients with factor VIII and IX inhibitors. PMID- 8236115 TI - The molecular basis of hemophilia A and the present status of carrier and antenatal diagnosis of the disease. PMID- 8236116 TI - Factor VIII structure and function. PMID- 8236117 TI - Factor XI deficiency. PMID- 8236118 TI - The role of factor XI in coagulation. PMID- 8236119 TI - Factor XI and platelets: activation and regulation. PMID- 8236120 TI - On the existence of platelet receptors for factor V(a) and factor VIII(a). AB - The surface of the platelet expresses procoagulant phospholipids that bind coagulation factors and contribute to the procoagulant activity of the cell. The evidence, however, also suggests that platelets may possess specific receptors for coagulation Factors V(a) and VIII(a). Examination of a limited set of the available data could lead to the controversial position that the procoagulant surface of platelets is defined exclusively by either procoagulant phospholipids, or specific receptors for the cofactors and proteases. At the present time insufficient data exist to exclusively support one or the other of these possibilities. Further work, however, should indicate clearly whether the putative receptors exist and what role they play, either alone or in concert with surface phospholipids, in the expression and regulation of platelet procoagulant activity. PMID- 8236121 TI - Platelet integrins. PMID- 8236122 TI - Clinical trials of platelet receptor inhibitors. PMID- 8236123 TI - von Willebrand factor: structure and function. PMID- 8236124 TI - The formation of the haemostatic plug--a special case of platelet aggregation. An experiment and a survey of the literature. AB - The formation of the haemostatic plug is an extremely fast process. This excludes, at least in its first phase, the involvement of soluble activating agents released from or produced by the platelets. An experiment with ADP activated, formaldehyde-fixed platelets shows that platelets with activated fibrinogen receptors will bind inactive platelets in the presence of fibrinogen and Ca(2+)-ions. A survey of the literature shows that platelet activation is accompanied by the clustering of the fibrinogen receptors. The surface of an activated platelet, which makes part of the growing haemostatic plug therefore is covered with patches of tightly packed fibrinogen. This allows the multisite combination with the statistically distributed low affinity receptors of the newly arriving platelets. Tightly packed fibrinogen, as present on clusters of the activated GP IIb/IIIa receptors as well as when absorbed to artificial surfaces acts as an activator of platelets. Thus, the propagation of the activation process is possible without a requirement for other, external activators. Such agents, which are released from platelets and, finally, thrombin formation, are nonetheless of vital importance, not for the formation but for the consolidation of the haemostatic plug. PMID- 8236125 TI - Treatment with tranexamic acid during pregnancy, and the risk of thrombo-embolic complications. AB - Tranexamic acid (AMCA) is an inhibitor of fibrinolysis used to treat fibrinolytic bleeding (e.g., menorrhagia and gastro-intestinal haemorrhage), and to prevent bleeding at surgery, in cases of abruptio placentae and general haemorrhage. As AMCA stabilises preformed clots and prolongs their dissolution, it has been debated whether treatment with AMCA might predispose to thrombosis by depressing the fibrinolytic system. Pregnant women constitute a group with low fibrinolytic capacity and an increased frequency of thrombosis further increased after Caesarean section, and are thus more likely to be susceptible to antifibrinolytic therapy. We therefore carried out a retrospective analysis of the case records of 2,102 patients with various bleeding disorders during pregnancy. Of the 256 patients treated with AMCA (mean duration of treatment, 46 days), 169 were delivered by Caesarean section. Of the remaining 1,846 patients (i.e., controls), 443 were delivered by Caesarean section. The relationship between the use of AMCA and the occurrence of thrombo-embolism was calculated with 95% confidence limits. Of the AMCA treated group (n = 256), two patients--one of whom belonged to the Caesarean section subgroup (n = 168)--had pulmonary embolism. Of the controls (n = 1,846), three patients had deep vein thrombosis and one had pulmonary embolism, all four cases belonging to the Caesarean section subgroup (n = 443). Thus, the findings in this high risk group of women with complicated pregnancies, frequently entailing delivery by Caesarean section, provided no evidence of any thrombogenic effect of AMCA. PMID- 8236126 TI - Effect of three fibrate derivatives and of two HMG-CoA reductase inhibitors on plasma fibrinogen level in patients with primary hypercholesterolemia. AB - In order to evaluate the effects of hypocholesterolemic drugs on plasma fibrinogen concentration, six groups of subjects with primary hypercholesterolemia have been put on treatment with diet alone or diet plus fenofibrate (100 mg t.i.d.), slow release bezafibrate (400 mg once a day), gemfibrozil (600 mg b.i.d.), simvastatin (20 mg once a day) or pravastatin (20 mg once a day) respectively. After 1 month of therapy, plasma fibrinogen significantly decreased by 9% and 15% in fenofibrate and bezafibrate groups respectively and increased by 19% in gemfibrozil treated patients. After 4 months of therapy the changes were -16% with fenofibrate, -10% with bezafibrate and +20% with gemfibrozil. No significant changes were observed in patients treated with diet alone, simvastatin or pravastatin. The fibrinogen lowering effect of fenofibrate and bezafibrate does not seem to be related to the hypolipidemic activity of the drugs. PMID- 8236127 TI - Protein S Tokushima: an abnormal protein S found in a Japanese family with thrombosis. AB - An abnormal protein S (PS) was found in a Japanese family with a high incidence of thrombosis. The proband is a woman who was born in Tokushima Prefecture. She had superior sagittal sinus thrombosis, thrombophlebitis of the left leg, and thrombosis of the placenta. She had a normal plasma level of free PS antigen but decreased PS activity. Her mother and aunt also had thrombophlebitis of the leg, and together with four other family members also showed a normal level but decreased activity of PS. This suggests that hereditary dysfunction of PS is inherited in this family as an autosomal dominant trait. The proband's PS appears to have a slightly higher molecular weight than normal PS both in the intact and modified form, suggesting that it has a molecular defect on the carboxyl-terminal side of the thrombin-sensitive site. This abnormal PS with apparently unique characteristics was named PS Tokushima. PMID- 8236128 TI - Replacement therapy with a purified protein C concentrate during initiation of oral anticoagulation in severe protein C congenital deficiency. AB - The case of an adult patient with moderately severe protein C deficiency (antigen 16%, activity 12%) is reported. Both parents had protein C levels compatible with heterozygous deficiency. Unlike other reported cases of severe protein C deficiency in adults, the onset of thrombotic symptoms occurred at 1 month of age; however, a symptom-free period until age 17 followed. Replacement therapy with a monoclonal antibody purified protein C concentrate was carried out during the initiation of oral anticoagulation after a course of i.v. heparin for deep vein thrombosis. The administration of the concentrate allowed maintenance of protein C above 50% until a stable therapeutic anticoagulation level could be obtained. This was reached within a short time, thus allowing safe administration of a loading dose of warfarin. We conclude that this approach to the prevention of skin necrosis seems more rapid and safer than previous schedules of oral anticoagulation in protein C-deficient patients. PMID- 8236129 TI - Positive association between self-reported fatty food consumption and factor VII coagulant activity, a risk factor for coronary heart disease, in 4246 middle-aged men. AB - Raised levels of factor VII coagulant activity (VIIc) have been reported to increase the incidence of CHD. Preliminary evidence from observational and experimental studies suggests that dietary fat intake is positively associated with VIIc. We explored this further in 4,246 men aged 45-69, who were found to be free of major CHD when screened for a primary prevention trial of antithrombotic medication. All men were asked about their consumption of fatty foods and changes in consumption in the last month. In the 9% of men who reported avoidance of fatty foods in the month before interview, age adjusted VIIc was 7.8% of standard (95% CI 5.1-10.6%) lower than in the remainder. Serum cholesterol and body mass index (BMI) were also significantly lower. The extent to which fat consumed in the past month had deviated from usual intake was significantly and positively related with VIIc, serum cholesterol and BMI. Thus, the VIIc difference between those eating much less fatty food than usual and those eating much more than usual was 11% of standard, with those eating their usual amount having an intermediate level. This study adds to the evidence that dietary fat intake influences VIIc and coagulability. The effect is rapid, so that much of the benefit of dietary fat reduction on thrombogenic risk in CHD is likely to occur within a short time. Thus, the results reinforce the value of a low fat diet, even in individuals with advanced atheroma, in whom dietary intervention has sometimes been considered unlikely to be effective. PMID- 8236130 TI - Thrombin production, inactivation and expression during open heart surgery measured by assays for activation fragments including a new ELISA for prothrombin fragment F1 + 2. AB - Activation of coagulation was studied during the peri-operative period in patients undergoing cardiopulmonary bypass (CPB) surgery using activation markers which have recently become available: prothrombin fragment F1 + 2 (F1 + 2), which is a measure of total thrombin generation, and thrombin-antithrombin complex, which is a measure of inactivation of free thrombin by antithrombin. Levels of the specific marker of fibrin breakdown, D-dimer, were also determined. F1 + 2 levels were assessed using a newly developed ELISA described herein which employs a neoantigen-specific capture antibody raised using a synthetic peptide; the latter antibody has been pre-adsorbed against prothrombin to ensure high specificity for F1 + 2. Increased generation of thrombin during surgery was clearly demonstrated despite maintenance of a high concentration of heparin during the period of extracorporeal blood circulation. There was a close association (r = 0.882) between the generation of thrombin (F1 + 2 levels) and its inhibition (TAT levels). Differences were noted, however, between the information provided by F1 + 2 and TAT, which are interpreted with regard to the different in vivo fates of F1 + 2 and thrombin. The enhanced activation and inhibition of coagulation observed during CPB was suppressed once physiological blood circulation was restored, with F1 + 2 returning to pre-surgical levels within 24 h after surgery. During the post-operative period D-dimer levels, which rose in concert with F1 + 2 and TAT levels, remained highly elevated, suggesting that not all of the generated thrombin was inactivated by antithrombin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236131 TI - Thrombin generation during cardiac surgery: is heparin the ideal anticoagulant? AB - Blood samples were collected from 43 patients undergoing elective cardiac surgery to determine the extent of thrombin generation and inhibition in patients when receiving heparin while undergoing cardiopulmonary bypass (CPB). Plasma prothrombin fragment F1 + 2 and thrombin-antithrombin III (TAT) levels were measured as markers of thrombin generation and inhibition, respectively. Both F1 + 2 and TAT levels increased significantly during the course of CPB despite the heparin causing significant systemic anticoagulation, i.e. the activated coagulation time (ACT) was prolonged to greater than 400 s throughout the entire surgical procedure. The extent of thrombin generation increased with time on CPB but did not differ between patients receiving normothermic and hypothermic cardioplegia during CPB. Furthermore, thrombin generation increased following the neutralization of the heparin with protamine sulphate, and continued to be elevated significantly 24 h post surgery. The observation that high dose heparin did not prevent thrombin generation during CPB, is consistent with previous experimental studies demonstrating that thrombin bound to fibrin or other surfaces (e.g. the CPB conduit) is resistant to antithrombin III/heparin inhibition, and thus able to facilitate further thrombin generation. The observation that thrombin generation continued to be elevated post surgery i.e. 24 h after neutralizing the heparin with protamine sulphate, suggests that the high dose heparin did not inhibit effectively all of the thrombin that had been generated. Thus, CPB patients may be at risk not only of bleeding and other side effects associated with the acute use of high dose heparin, but may also be at risk of further thrombosis-related events either acutely or chronically. PMID- 8236132 TI - Congenital histidine-rich glycoprotein deficiency. AB - The proband, a 43-year-old woman, suffered from right transverse sinus thrombosis during oral contraceptive treatment. A month after stopping the drug, her plasma activities of antithrombin III, protein C, protein S, heparin cofactor II, plasminogen and plasminogen activator inhibitor were normal, but her plasma histidine-rich glycoprotein (HRG) level was only 21% of the normal level of 109.5 +/- 51.5% (mean +/- 2 SD). The HRG concentrations in her plasma determined on four different occasions over 6 months were similar. She showed no clinical signs of liver insufficiency or sepsis. Low levels of plasma HRG (20% to 35% of normal) were also found in her aunt, uncle and two daughters. These results suggest that congenital HRG deficiency is inheritary in this family. PMID- 8236133 TI - Features of thrombi and diagnostic accuracy of impedance plethysmography in symptomatic and asymptomatic deep vein thrombosis. AB - Impedance plethysmography (IPG) has high sensitivity and specificity in patients with symptomatic deep vein thrombosis (DVT) while it fails to detect asymptomatic DVT. The aim of this study was to determine whether the features of thrombi such as location, size and occlusiveness could explain the different accuracy of IPG in symptomatic and asymptomatic DVT patients. One-hundred and seventeen consecutive outpatients with a clinical suspicion of DVT and 246 consecutive patients undergoing hip surgery were admitted to the study. In symptomatic patients IPG was performed on the day of referral, followed by venography, while in asymptomatic patients IPG was performed as a surveillance programme, followed by bilateral venography. A venography proved DVT was observed in 37% of the symptomatic patients and 34% of the asymptomatic limbs. A significantly higher proportion of proximal DVTs was found in symptomatic patients than in asymptomatic patients (78% vs 46%; p = 0.001). The mean Marder score, taken as an index of thrombus size, was significantly higher in symptomatic patients than in asymptomatic patients (19.0 vs 9.6; p = 0.0001). A significantly higher proportion of occlusive DVTs was observed in symptomatic than in asymptomatic patients (69% vs 36%; p = 0.001). We conclude that the unsatisfactory diagnostic accuracy of IPG in asymptomatic DVT is due to the high prevalence of distal, small and non occlusive thrombi. Such thrombi are unlikely to cause a critical obstruction of the venous outflow and therefore to produce a positive IPG. PMID- 8236134 TI - Pharmacokinetics of monoclonally-purified and recombinant factor VIII in patients with severe von Willebrand disease. AB - A monoclonally-purified factor VIII (FVIII) concentrate, containing little von Willebrand factor (vWF), was infused to 11 patients with severe von Willebrand disease and unmeasurable levels of plasma vWF. In comparison with the historical data obtained infusing hemophiliacs in the same conditions, monoclonally-purified FVIII had a significantly shorter half-life and faster clearance from plasma but similar in vivo recovery and volume of distribution. Two additional patients with severe von Willebrand disease were also infused with recombinant FVIII totally devoid of vWF. Half-life was very short and in vivo recovery low, with a larger volume of distribution than for monoclonally-purified FVIII. We conclude that in patients with severe von Willebrand disease the small amounts of vWF contained in the monoclonally-purified FVIII concentrate are not sufficient to stabilize infused FVIII, nor to support the normal circulation of endogenous FVIII that these patients produce at a normal rate. PMID- 8236135 TI - Protein C activity in healthy volunteers--influence of age, sex, smoking and oral contraceptives. AB - Interpretation of protein C (PC) levels in a given individual has several limitations. A normal PC activity does not necessarily exclude a genetic deficiency nor can a reduced level confirm it. Measuring PC amidolytic activity in 9,648 healthy blood donors has allowed identification of demographic factors which cause variation in PC activity and further hinder interpretation. PC activity displays a log normal distribution and significant variation with age. This is most marked in young adult males when mean PC activity rises from 0.86 iu/ml (15-19 years) to 1.04 iu/ml (45-49 years; P < 0.0001). Pre-menopausal females, who for most age ranges, have mean PC activity below their male contemporaries, show a less marked rise with age until the menopause when PC activity rises further. The use of hormonal contraceptive preparations is associated with an increase in mean PC activity of 0.05-0.08 iu/ml while smoking habit has no influence on PC activity. In view of these findings we strongly recommend the use of age and sex restricted reference ranges when interpreting PC activity. PMID- 8236136 TI - Effects of hirudin on activated partial thromboplastin time determined with ten different reagents. AB - Hirudin prolongs the APTT when added to normal plasma and the extent of prolongation depends on the type of reagent used. The aim of this study was to compare the dose-response curves of 10 widely used APTT reagents for linearity and parallelism. On each of 10 working days a normal pooled plasma was mixed with increasing amounts of recombinant hirudin (HBW023) ranging from 0 to 5 micrograms/ml and tested for APTT by photo optical coagulometer. Within each working day, clotting times were measured in duplicate and the order of testing with each reagent was changed every day. Results were expressed as ratios of clotting times with hirudin to clotting times without hirudin, and the values plotted against the hirudin concentration on a log-log scale. The dose-response curves for all reagents were linear over 0.3-1.2 micrograms/ml. The reagent related slopes ranged from 0.225 +/- 0.003 to 0.303 +/- 0.003 (mean +/- SE) and were significantly different. Precision studies indicated that the least sensitive reagent was also the least precise. These findings indicate that the clotting time values obtained for patients treated with hirudin will vary depending on the APTT reagent used. PMID- 8236137 TI - On the mechanism of coagulation inhibition on surfaces with end point immobilized heparin. AB - A well established technique to improve blood compatibility of artificial materials for use in the circulation is to coat the surface with heparin. The present report describes the antithrombin mediated inhibition of thrombin and factor Xa by surfaces modified with end point immobilized heparin. The reaction was followed by conventional chromogenic substrate based enzyme assays as well as by immunological measurement of the enzyme inhibitor (thrombin-antithrombin) complex formation. Both enzymes were rapidly inactivated by heparin surfaces after selective presaturation with antithrombin on the immobilized high affinity heparin molecules. The thrombin inhibitory capacity was enhanced when both high and low affinity heparin were preadsorbed with inhibitor. The main part of the thrombin-antithrombin complex formed remained bound to the surface, however, without functionally blocking the activity of the high affinity sequence of the immobilized heparin. Aliquots of recalcified plasma were slowly rotated in loops of heparinized tubing to investigate whether the main thromboresistant function of the surface was exerted at the level of thrombin or by inactivation of preceding enzymes. After 1 h no visible clotting occurred and only trace amounts of thrombin (0.07 IU/ml), measured as thrombin-antithrombin complexes, had been formed. In non-heparinized loops and in the test tube plasma clotted after 20 min. The thrombin generation when clotting occurred was in the order of 10 IU/ml. It is concluded that the immobilized heparin mediates inhibition of the coagulation cascade prior to prothrombin activation. PMID- 8236138 TI - Thrombolytic and haemorrhagic effects of bolus doses of tissue-type plasminogen activator and a hybrid plasminogen activator with prolonged plasma half-life (K2tu-PA: CGP 42935). AB - K2tu-PA is a hybrid plasminogen activator linking the kringle 2 domain of tissue type plasminogen activator (t-PA) to the catalytic protease domain of single chain urokinase-type plasminogen activator (scu-PA). K2tu-PA, as t-PA has high affinity for fibrin and is activated by fibrin but has a longer plasma half-life (over 30 min). The aim of this study was to compare the effects of bolus doses of recombinant t-PA (rt-PA) and K2tu-PA, on: 1) lysis of performed thrombi (fibrinolysis), 2) accretion of new fibrin on pre-existing thrombi during fibrinolysis (thrombus growth), 3) thrombolysis as assessed by reduction of thrombus weight and 4) systemic plasma proteolysis and blood loss from a standard wound. A jugular vein thrombosis model and an ear bleeding model were adopted in rabbits. Saline produced 11 +/- 2% fibrinolysis. rt-PA, 0.2 mg/kg, 0.4 mg and 0.8 mg/kg produced 35 +/- 4%, 54 +/- 4% and 78 +/- 6% fibrinolysis, respectively. K2tu-PA, at the same doses, produced 39 +/- 5%, 57 +/- 6% and 83 +/- 6% fibrinolysis, respectively. Thus, no differences in the fibrinolytic activity of rt-PA and K2tu-PA were observed. Injection of saline was followed by an accretion of 56.4 +/- 5.9 micrograms of radioactive new fibrin on the thrombi. The injection of the three increasing doses of rt-PA was followed by an accretion of 54.9 +/- 5.3 micrograms, 49.1 +/- 6.1 micrograms and 47.2 +/- 4.8 micrograms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236139 TI - The roles of alpha 2-antiplasmin and plasminogen activator inhibitor 1 (PAI-1) in the inhibition of clot lysis. AB - The relative importance of the two major inhibitors of fibrinolysis, alpha 2 antiplasmin (alpha 2-AP) and plasminogen activator inhibitor (PAI-1), were investigated using a simple microtitre plate system to study fibrin clot lysis in vitro. Cross-linked fibrin clots contained plasminogen and tissue plasminogen activator (t-PA) at concentrations close to physiological. Purified alpha 2-AP and PAI-1 caused dose-dependent inhibition. All the inhibition due to normal plasma, either platelet-rich or poor, was neutralised only by antibodies to alpha 2-AP. Isolated platelets, at a final concentration similar to that in blood, 2.5 x 10(8)/ml, markedly inhibited clot lysis. This inhibition was neutralised only by antibodies to PAI-1. At the normal circulating ratio of plasma to platelets, alpha 2-AP was the dominant inhibitor. When the platelet:plasma ratio was raised some 20-fold, platelet PAI-1 provided a significant contribution. High local concentrations of PAI-1 do occur in thrombi in vivo, indicating a role for PAI-1, complementary to that of alpha 2-AP, in such situations. PMID- 8236140 TI - A slow clearing, fibrin-specific, PAI-1 resistant variant of t-PA (T103N, KHRR 296-299 AAAA). AB - Site directed mutagenesis was used to construct a t-PA variant that contains an additional glycosylation site in the first kringle domain (T103N) combined with a tetra-alanine substitution in the protease domain (KHRR 296-299 AAAA). This combination variant has a plasma clearance rate that is 4.5-fold slower in rats and 5.4-fold slower in rabbits than t-PA. It is also less than one tenth as active as t-PA towards plasminogen in the presence of fibrinogen, and has approximately twice the normal activity in the presence of fibrin. It shows substantial resistance to the fast acting inhibitor, plasminogen activator inhibitor-1 (PAI-1), requiring a 10-fold greater molar excess of PAI-1 to reduce its activity by 50%, compared to t-PA. This is the result of a reduction of nearly 100-fold in the second order rate constant for PAI-1 inactivation. These results show that it is possible to combine mutations in different domains of t PA to construct a variant which is simultaneously slower clearing, less reactive towards plasminogen in the absence of a fibrin clot, and resistant to inactivation by PAI-1. PMID- 8236141 TI - A variant of t-PA (T103N, KHRR 296-299 AAAA) that, by bolus, has increased potency and decreased systemic activation of plasminogen. AB - In the accompanying paper, we reported that the properties of decreased plasma clearance rate, increased fibrin specificity, and resistance to inactivation by PAI-1 could be effectively combined in the t-PA variant T103N, KHRR 296-299 AAAA. In the current study we evaluated the in vivo efficacy of this variant as well as variants containing the individual mutations T103N and KHRR 296-299 AAAA. Plasma clearance and in vivo lysis of whole blood and platelet-rich clots were determined in a rabbit arterio-venous shunt model. The T103N containing variants were administered as an intravenous (i.v.) bolus. KHRR 296-299 AAAA and t-PA were infused i.v. over 90 min. The clearance rate of the KHRR 296-299 AAAA variant was similar to t-PA. However, the clearance of the T103N and T103N, KHRR 296-299 AAAA variants were 8 and 6-fold reduced, respectively. Potency of the variants relative to t-PA on whole blood clots ranged from 0.9 (T103N, KHRR 296-299 AAAA) to 1.7 (T103N). Relative potency on platelet-rich clots ranged from 2.4 (T103N) to 4.2 (T103N, KHRR 296-299 AAAA). Fibrinogen concentrations in rabbits 120 min after dosing with a 2.5 mg/kg bolus were: 24, 16, 82, and 77% of initial for t PA; T103N; KHRR 296-299 AAAA; and T103N, KHRR 296-299 AAAA treatment groups, respectively. These results suggest that the T103N, KHRR 296-299 AAAA variant of t-PA, given as a bolus, could result in greater efficacy, particularly on refractory platelet-rich clots, without inducing the severe systemic lytic state produced by a bolus of a less fibrin specific variant. PMID- 8236142 TI - Pharmacokinetics of saruplase, a recombinant unglycosylated human single-chain urokinase-type plasminogen activator and its effects on fibrinolytic and haemostatic parameters in healthy male subjects. AB - Pharmacokinetics of two doses of the recombinant single-chain urokinase-type plasminogen activator (r-scu-PA) saruplase (40 and 20 mg) and its effect on fibrinolytic and haemostatic parameters were studied in six healthy male subjects using a randomized, double-blind, placebo-controlled, cross-over study. Special precautions were taken to prevent artefactual in vitro effects on fibrinolytic activity. The clearance of saruplase ranged from 310 to 862 ml/min and the apparent volume of distribution of the central compartment was about 8 1. Both doses of saruplase caused alpha 2-antiplasmin consumption, indicating some systemic fibrinolytic activation. However, the 20 mg dose caused no detectable fibrinogen breakdown and only a small increase in total fibrin/fibrinogen degradation products (TDP) (from 0.16 microgram/ml [range 0.14 to 0.19] to 0.78 microgram/ml [range 0.56 to 1.26]), while the 40 mg dose produce a fibrinogen breakdown to an average value of 44% (range 19 to 60%) and TDP increased from 0.12 microgram/ml (range 0.11-0.12) to 2.29 micrograms/ml (range 0.45 to 5.55). The breakdown of fibrinogen was related to the quantity of saruplase converted to active two-chain u-PA (tcu-PA) in vivo (6 to 22% conversion). There were no important effects of saruplase on overall blood coagulation (activated partial thromboplastin time) and platelet function (collagen induced platelet aggregation, urinary [2,3-dinor]-thromboxane B2 excretion and plasminogen activator inhibitor 1 [PAI-1] release from platelets). Saruplase is cleared rapidly from the plasma and a variable amount is converted to tcu-PA. This two chain form of u-PA probably causes the dose-dependent systemic fibrinolytic activation. PMID- 8236143 TI - Functional properties of p-anisoylated plasmin-staphylokinase complex. AB - The kinetic and fibrinolytic properties of a reversibly acylated stoichiometric complex between human plasmin and recombinant staphylokinase (plasmin-STAR complex) were evaluated. The acylation rate constant of plasmin-STAR by p amidinophenyl-p'-anisate-HCl was 52 M-1 s-1 and its deacylation rate constant 1.2 x 10(-4) s-1 (t1/2 of 95 min) which are respectively 50-fold and around 3-fold lower than for the plasmin-streptokinase complex. The acylated complex was stable as evidenced by binding to lysine-Sepharose. However, following an initial short lag phase, the acylated plasmin-STAR complex activated plasminogen at a similar rate as the unblocked complex, whereas the acylated plasmin-streptokinase complex did not activate plasminogen. These findings indicate that STAR, unlike streptokinase, dissociates from its acylated complex with plasmin in the presence of excess plasminogen. In agreement with this hypothesis, the time course of the lysis of a 125I-fibrin labeled plasma clot submerged in citrated human plasma, is similar for acylated plasmin-STAR, unblocked plasmin-STAR and free STAR (50% clot lysis in 2 h requires 12 nM of each agent). The plasma clearances of STAR-related antigen following bolus injection in hamsters were 1.0 to 1.5 ml/min for acylated plasmin-STAR, unblocked plasmin-STAR and free STAR, as a result of short initial half-lives of 2.0 to 2.5 min. The dissociation of the anisoylated plasmin-STAR complex and its consequent rapid clearance suggest that it has no apparent advantages as compared to free STAR for clinical thrombolysis. PMID- 8236144 TI - Effect of diltiazem and low-dose aspirin on platelet aggregation and ATP release induced by paired agonists. AB - The authors studied the effects of diltiazem, administered alone and together with low-dose aspirin, on the platelet response to paired agonists. After a baseline period, 25 healthy volunteers were given oral diltiazem for 1 week (120, 240, or 360 mg/day), and then crossed over randomly between 1 week on diltiazem plus aspirin (81 mg/day), and 1 week on aspirin (81 mg/day) alone. Platelet function was tested on 2 consecutive days in each period. Synergistic platelet aggregation and ATP release were obtained at baseline using a subthreshold concentration of arachidonic acid combined with platelet activating factor, ADP, or epinephrine. Diltiazem resulted in significant decrease from baseline in platelet aggregation and ATP release using the arachidonic acid-epinephrine combination (35% and 40% decrease, respectively, p < 0.01) and a significant decrease in aggregation using the arachidonic acid-ADP combination (22% decrease, p < 0.01). The effects were neither dose-related, nor accompanied by any significant change in serum thromboxane B2 levels or bleeding times. There was no significant difference between the effects of aspirin alone and aspirin plus diltiazem on the synergistic platelet aggregation and ATP release induced by the paired agonists, or on thromboxane B2 levels or bleeding times. Diltiazem administered in vivo partially inhibits the synergistic platelet aggregation and ATP release induced by paired agonists; however, in contrast to a previous in vitro study it does not potentiate the platelet-inhibitory effect of aspirin. PMID- 8236145 TI - Platelet activating properties of murine monoclonal antibodies to beta 2 glycoprotein I. AB - Previously developed murine monoclonal antibodies (MAbs) to human beta 2 glycoprotein I (beta 2 GPI), a plasma protein required for the binding of anti phospholipid antibodies, were studied for anti-platelet reactivity and influence on platelet function. The six MAbs (IgG1 isotype) tested interacted with both intact and fixed platelets in a beta 2 GPI-dependent manner. Carbamylated beta 2 GPI was still recognized by MAbs but was unable to mediate platelet-antibody binding. MAbs induced aggregation and secretion responses of platelets in platelet-rich plasma (PRP) and whole blood, provided subthreshold concentrations of weak agonists (i.e. ADP or adrenaline) were added. When aggregation in PRP was evaluated by a counting technique instead of turbidometrically, the sole addition of MAbs led to a rapid fall in single platelets. Triggering gel-filtered platelets with MAbs together with beta 2 GPI, but not its carbamylated form, led to platelet activation after a lag time, as monitored by aggregometry, measurements of ATP and beta-thromboglobulin secretion and calcium mobilization. F(ab')2 fragments of one of the MAbs failed to activate platelets but inhibited the responses to the whole antibody. This process thus depends on MAbs binding to platelets through both Fab and Fc domains, as confirmed by the suppression of platelet responses upon pretreatment with the anti-Fc gamma RII MAb IV.3. Aggregation and secretion induced by MAbs plus beta 2 GPI did not require exogenous fibrinogen and were variably inhibited in the presence of acetyl salicylic acid, apyrase or Ca2+, depending on the concentrations used for the two proteins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236146 TI - Anticardiolipin antibodies block the inhibition by beta 2-glycoprotein I of the factor Xa generating activity of platelets. AB - Antiphospholipid antibodies, defined either by lupus anticoagulant (LA) activity or positive anticardiolipin immunoabsorbent assay (ACA) are associated with a predisposition to thromboses, recurrent fetal loss or thrombocytopenia. The mechanisms for these predispositions remain undefined. We have enriched immunoglobulin fractions from two patient plasmas to obtain antibodies with LA activity but no ACA, or conversely, with ACA positivity but no LA, in order to investigate in vitro characteristics which might explain a thrombotic propensity. beta 2-glycoprotein I (beta 2-GPI), the plasma cofactor required for ACA binding to negatively charged phospholipid, has previously been shown to inhibit prothrombinase generation in the presence of activated platelets (8). We now report that beta 2-GPI, at physiological concentrations, inhibits the generation of factor Xa in the presence of activated gel-filtered platelets. Further, ACA interferes with this inhibition, resulting in protracted, unopposed factor Xa generation. This interference with beta 2-GPI, a natural anticoagulant component of plasma, is potentially prothrombotic. LA immunoglobulins behave differently and inhibit factor Xa generation in a manner similar to beta 2-GPI. These findings provide the basis for a previously unsuspected mechanism for thrombosis in patients with aPL. PMID- 8236147 TI - Standards for assay of von Willebrand factor concentrates: a collaborative study. AB - A multilaboratory collaborative study was undertaken to assess the feasibility of using a plasma standard for expressing the results of assays for the von Willebrand factor content of von Willebrand factor concentrates and of factor VIII concentrates. Thirteen laboratories tested six concentrates for von Willebrand factor antigen, ristocetin cofactor activity, and multimer content using the World Health Organization plasma standard for factor VIII/von Willebrand factor, 87/718, as a standard. Only a few assays were invalid because of nonparallelism or nonlinearity. Significant interlaboratory and interassay differences were found for both von Willebrand factor antigen and ristocetin cofactor activity. There was generally good agreement between the laboratories with respect to the multimer content in the preparations. With respect to assay validity, a plasma standard could be suitable for assaying concentrated preparations of von Willebrand factor. PMID- 8236148 TI - Registry of clinical trials of antithrombotic drugs in cancer: second report. The Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis Subcommittee on Hemostasis and Malignancy. PMID- 8236149 TI - Antithrombin III mutation database: first update. For the Thrombin and its Inhibitors Subcommittee of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis. PMID- 8236150 TI - A deletion located in the 3' non translated part of the factor IX gene responsible for mild haemophilia B. PMID- 8236151 TI - International sensitivity or insensitivity indexes for calibrating working methods against a reference procedure. PMID- 8236152 TI - rHuEPO increases urinary excretion of fibrin degradation products in haemodialysed patients. PMID- 8236153 TI - Further evidence that alpha-granule components such as platelet factor 4 are involved in platelet-IgG-heparin interactions during heparin-associated thrombocytopenia. PMID- 8236154 TI - Clot retraction increases clot resistance to fibrinolysis by condensing alpha 2 plasmin inhibitor crosslinked to fibrin. PMID- 8236155 TI - The antithrombotic and anticoagulant effects of a synthetic tripeptide and recombinant hirudin in various animal models. AB - The pharmacologic activities of two thrombin inhibitors (D-MePhe-Pro-Arg-H, recombinant-hirudin) were compared in two animal models. The antithrombotic effect was investigated in vivo in rabbits using a modified Wessler stasis thrombosis model. During these experiments, blood was drawn for ex vivo testing to determine the coagulation profile and to determine plasma concentrations using pre-constructed calibration curves. A dose-dependent antithrombotic effect was observed for both agents. On an equigravimetric basis (100 micrograms/kg i.v.), r hirudin showed a stronger antithrombotic effect than the tripeptide, which correlated well with the ex vivo anticoagulant effect. No adverse reactions were observed during this study. In a rabbit ear bleeding model, a dose and time dependent hemorrhagic effect was observed for both agents. Only slight bleeding effects were observed at 1.0 mg/kg dosages. These studies show that the tripeptide D-MePhe-Pro-Arg-H and r-hirudin are specific thrombin inhibitors with potent antithrombotic effects and a high therapeutic (antithrombotic/hemorrhagic) index. Furthermore, the results of these two animal models and ex vivo analyses can be used to determine the therapeutic index of thrombin inhibitors. PMID- 8236156 TI - Thermal stability of low molecular weight urokinase during heat treatment. I. Effects of protein concentration, pH and ionic strength. AB - Exposure of low molecular weight urokinase (LMW-UK) to prolonged heating (60 degrees C, 10 hours) is used to inactivate possible viral contaminants. This process leads to a significant loss of active enzyme. Amidolytic activity was monitored following heat treatment in order to establish the conditions for maintaining the optimal stability of LMW-UK. The effects of pH, ionic strength, protein concentration, and various ionic additives were examined. While LMW-UK is stable across a wide pH range (pH 2-11), heating LMW-UK in aqueous solution leads to complete loss of activity except between pH 4 and 7.5. The mechanism of inactivation was delineated using activity assays as well as turbimetric and spectroscopic methods. Thermal inactivation occurs via aggregation of unfolded LMW-UK, followed by subsequent precipitation. Threshold effects upon the thermally-induced aggregation of LMW-UK were observed. PMID- 8236157 TI - Effect of leucocyte products on platelet thrombus formation, coagulation and spontaneous thrombolysis, as measured from native human blood, in vitro. AB - Contribution of leucocytes to formation and lysis of arterial (platelet) thrombi was investigated. Secretory products of polymorphonuclear leucocytes (PMNs) during phagocytosis and cell lysates were prepared from eight volunteers. Platelet-rich thrombi were formed in flowing native human blood either by shear stress or by collagen fibre, by haemostatometry. Tested in eight volunteers, PMN products significantly enhanced both thrombotic reactions and induced lysis of these thrombi. A specific inhibitor of leucocyte proteases (eglin c) inhibited platelet reaction to shear-stress and collagen and spontaneous thrombolysis. Our findings provide further evidence for the prothrombotic and potent thrombolytic effect of leucocytes associated with an arterial thrombus. PMID- 8236158 TI - The potential role of platelet PAl-1 in t-PA mediated clot lysis of platelet rich plasma. AB - The potential role of platelets in platelet rich plasma clot lysis induced by tissue plasminogen activator (t-PA) was investigated. At the various concentrations of both single chain t-PA (sct-PA) and two chain t-PA (tct-PA) (1.5nM, 3nM, and 6nM), we compared the t-PA mediated lysis time of platelet rich plasma clot (PRP-clot) with that of platelet poor plasma clot (PPP-clot). At the concentrations ranged from 1.5 to 6 nM of both types of t-PA, the clot lysis time of PRP-clot was longer than that of PPP-clot. This elongation was more significant in the tct-PA induced clot lysis than that in the sct-PA induced clot lysis. At the concentration of 3nM of tct-PA, the lysis time of PRP-clot was longer by a factor of 30% in comparison with that of PPP-clot. When the release and the aggregation of platelets were blocked by prostaglandin E1 (PGE1) and theophylline in this experiment, the lysis time of PRP-clot was essentially the same as that of PPP-clot. We then measured the antigen levels of total PAI-1 and t-PA-PAI-1 complex in the lyzed solutions of PRP-clot and PPP-clot to analyse the possible effect of plasminogen activator inhibitor-1 (PAI-1) present in platelets. Most of PAI-1 in a PPP-clot lyzed sample existed as t-PA-PAI-1 complex. In the lyzed solution of PRP-clot, however, the antigen levels of both total PAI-1 and t-PA-PAI-1 complex were significantly higher than those in PPP clot, and larger amounts of PAI-1 existed as free PAI-1 which possesses activity. These data suggest that at least certain amounts of PAI-1 in platelets exist as an active form and inhibits t-PA activity resulting in the prolongation of the clot lysis time. Activation of platelets, therefore, seems to play an important role in the platelet rich plasma clot lysis induced by t-PA. PMID- 8236159 TI - Mouse antithrombotic assay. Inhibition of platelet thromboembolism by disintegrins. AB - The mouse antithrombotic assay represents a model of fatal pulmonary thromboembolism induced by intravenous injection of collagen and epinephrine. Mice were protected by low doses of two disintegrins, albolabrin (10 micrograms/mouse) and eristostatin (0.6 micrograms/mouse), whereas high doses of a thrombin inhibitor and an inhibitor of von Willebrand Factor binding to glycoprotein Ib were not effective. Injection of collagen and epinephrine resulted in the drop of platelet count and accumulation of platelet aggregates in the lung that appears to be the immediate cause of death. Albolabrin or eristostatin administration did not prevent the decrease of platelet count. Injection of albolabrin resulted in the formation of smaller and reversible platelet aggregates in the lungs and decreased accumulation of 51Cr-labeled platelets in the lung suggesting that this disintegrin decreases formation of platelet aggregates in vivo. We compared the effects of albolabrin and erisostatin on platelet aggregation, tail bleeding time, and survival of challenged animals. Eristostatin was about 5 times more potent in inhibiting platelet aggregation in vitro than albolabrin and 38 times more potent than albolabrin in protecting animals from sudden death. Both disintegrins, at the same doses (0.6-5 micrograms/mouse), caused similar dose-dependent prolongation of the bleeding time; however, only eristostatin exerted a protective effect. In conclusion, a) the mouse antithrombotic assay is a suitable model to screen and to evaluate the potency of platelet fibrinogen receptor antagonists in vivo; b) the results of the antithrombotic assay correlate better with the inhibition of platelet aggregation in vitro than with the prolongation of bleeding time. PMID- 8236160 TI - Antithrombotic activity of recombinant tick anticoagulant peptide and heparin in a rabbit model of venous thrombosis. AB - An in vivo rabbit model of venous thrombosis which includes physiological blood flow was used to compare the efficacy of the potent and specific factor Xa inhibitor recombinant tick anticoagulant peptide (rTAP) with standard heparin in the prevention of venous thrombus formation. In anesthetized rabbits, an autologous thrombus was induced with thrombin in a jugular vein and the increase in thrombus size was determined by measuring the accretion of intravenously injected [125I]fibrin(ogen) onto the developing thrombus. The effects of rTAP on hemostasis were monitored by changes in APTT values and template bleeding times. Inhibition of thrombus formation by an intravenous bolus followed by infusion of either rTAP or heparin exhibited a dose-response relationship with an IC50 of 0.9 micrograms/kg/min and 0.12 units/kg/min, respectively. At the IC50 doses, both rTAP and heparin inhibited fibrin(ogen) deposition without any significant effect on APTT or bleeding times. Bleeding times were modestly elevated at the fully efficacious doses of rTAP and heparin. Significant changes in APTT (1.9 +/- 0.3 fold over baseline) were only evident at the highest dose of rTAP while heparin caused a significant dose-dependent increase from 1.3 +/- 0.2 to greater than 4.2 +/- 0.6 fold over baseline. Therefore, in this rabbit model of venous thrombosis, specific inhibition of factor Xa by rTAP is an effective antithrombotic mechanism that does not require changes in systemic hemostatic parameters. PMID- 8236161 TI - Antithrombotic effect of recombinant human soluble thrombomodulin on endotoxin induced disseminated intravascular coagulation in rats. AB - Thrombomodulin (TM) is an endothelial cell membrane glycoprotein which neutralizes thrombin procoagulant activity and accelerates the thrombin-catalyzed activation of protein C. We expressed recombinant human soluble TM (rhs-TM) in Chinese hamster ovary cells and compared the effects of rhs-TM and heparin on endotoxin-induced experimental disseminated intravascular coagulation (DIC) in rats. Experimental DIC was induced by a continuous intravenous infusion of endotoxin for four hours. rhs-TM or heparin was infused simultaneously with endotoxin. Treatment with rhs-TM significantly reversed the endotoxin-induced changes in significantly reversed the endotoxin-induced changes in following parameters: platelet count, fibrinogen level and fibrinogen and fibrin degradation products. Furthermore, glomerular fibrin deposits elevated by endotoxin treatment were reduced by the rhs-TM administration. Heparin showed the similar effects to rhs-TM. Activated partial thromboplastin time (APTT) in rats receiving rhs-TM were slightly longer than APTT in endotoxin-treated rats, but rats receiving heparin had much more prolonged APTT. From these results, we concluded that rhs-TM may be useful for the clinical treatment of DIC while having only minor adverse effects on APTT. PMID- 8236162 TI - Homocysteinemia: association of a metabolic disorder with vascular disease and thrombosis. PMID- 8236163 TI - Effects of Poloxamer 188 on fibrin network structure, whole blood clot premeability and fibrinolysis. AB - The effects of Poloxamer 188 (0-5 mg/ml) on the permeability, turbidity, compaction, and fibrinolysis of fibrin network developed in human plasma, and on the permeability and fibrinolysis of network developed in whole blood were examined. Poloxamer 188 was found to increase network permeability and compaction in plasma. In networks in plasma, effects on the fibre mass-length ratio from turbidity and fibrinolysis with recombinant tissue plasminogen activator were small. Poloxamer did not alter the fibrinolysis with streptokinase. The increase in fibrin network permeability at low poloxamer concentrations was not attributable to an increase in fibre thickness, but results from alterations in the arrangement of fibrin fibres. Poloxamer also significantly increased the permeability of networks developed in whole blood. Studies with the platelet inhibitor cytochalasin B demonstrated that this effect in whole blood networks was partly from facilitation of platelet induced clot retraction. Poloxamer was not found to affect streptokinase induced fibrinolysis of whole blood networks. The effects of poloxamer support the hypothesis that depletion flocculation of fibrin intermediaries by soluble macromolecules is a significant determinant of network permeability. The therapeutic use of poloxamer will result in altered fibrin function in particular its permeability and mechanical stability. These alterations may contribute to its described antithrombotic and rheological effects. PMID- 8236164 TI - Molecular cloning of the mouse equivalent of CD9 antigen. AB - The CD9 antigen was originally described as a 24 kDa molecule present on B lineage-derived acute lymphoblastic leukemia cells and developing B-lymphocytes. However, platelets express a large amount of CD9 antigen and can be activated by CD9 antibodies. We report here the cloning and sequencing of a cDNA coding for the mouse CD9 antigen. There is 89% homology at amino acid level between the human and mouse CD9 molecules. Most of the differences (19 out of 24) are located in the large putative extracellular domain encoded by exons 5 and 6. CD9 antigen belongs to a new cell surface protein family which includes TAPA1 and the platelet activation antigen CD63. These proteins share common structural features with the CD9 antigen and a similar distribution of the evolutionarily variable region. PMID- 8236165 TI - The effects of calpeptin (a calpain specific inhibitor) on agonist induced microparticle formation from the platelet plasma membrane. AB - Platelets activated by various agonists produce formation of vesicles shed from the plasma membrane (microparticles). However, the mechanism of microparticle (MP) formation has not been clarified yet. The aim of the present study was to determine the possibility of involvement of calpain (a Ca(2+)-dependent thiol protease) in MP formation. Washed platelets preincubated with calpeptin, a cell permeable calpain specific inhibitor, or with a vehicle were activated by thrombin plus collagen or by calcium ionophore A23187. Flow cytometry was used to detect the amount of microparticle formation by using murine monoclonal antibodies against GP IIb-IIIa or GP IIb and fluorescein 5-isothiocyanate labeled goat anti-mouse IgG. MP formation stimulated either by thrombin plus collagen or by A23187 was inhibited by calpeptin in a dose dependent manner. The microparticle formation from platelets activated by A23187 reached a plateau in approximately 5 min after activation, whereas that from platelets activated by thrombin plus collagen reached a plateau at 30 min following the stimulation. These time sequences corresponded well with those of degradation of actin-binding protein (ABP), a well known substrate of calpain, of platelets activated by these two stimulations. However, the inhibition of MP formation by calpeptin was more marked in the early stage (within 10 min) than in the late stage (after 30 min) of platelet activation. At 30 min after platelet activation by either two stimulations, a significant amount of microparticle formation was observed in the presence of 30 microM calpeptin, which inhibited hydrolysis of ABP almost completely. Our data suggest the involvement of calpain in the early stage (especially within 10 min) of microparticle formation. PMID- 8236166 TI - Two-year follow-up of aspirin responder and aspirin non responder. A pilot-study including 180 post-stroke patients. AB - Aspirin is proposed to be effective in stroke-prophylaxis because it completely inhibits the platelet prostanoid-pathway. In about 90% of stroke victims, increased platelet reactivity (PR) can be reduced to the normal range by aspirin. Twelve hours later, about one third of them show an enhanced PR again. These patients are called secondary aspirin non responders (SANR). In this study the potential pathogenetic and prognostic impact of this biological feature on stroke recurrence was evaluated. Before discharge from the hospital, PR was determined 12 hours after an oral administration of 500 mg aspirin in 180 patients aged 58 +/- 15 years; 74 were female and 106 male. All had suffered a stroke in the internal carotid artery territory. Patients were treated with 3 x 500 mg aspirin/d and were followed up over a 24-month period. Major endpoints of this study were stroke, myocardial infarction or vascular death. On discharge from the hospital, 120 of the 180 patients showed a normal PR under aspirin treatment. High test values were found in 60 patients (SANR). Six patients were lost for follow-up. Because of side effects 36 (20%) of the 180 patients enrolled discontinued medication. Major endpoints occurred in 4 of these 36 patients (11%) and in 25 of the 138 remaining patients (18.1%); 19 patients died in consequence of a vascular event during the observation period. Major endpoints were seen in only 5 of 114 (4.4%) of the aspirin responders, but in 24 out of 60 SANR (40%, p < 0.0001). It may be assumed that early identification of SANR's is a clinically useful tool to classify patients at high risk for recurrence of vascular events. PMID- 8236167 TI - Changes in fibrinolytic parameters in male patients with type 2 (non-insulin dependent) diabetes mellitus. AB - We examined changes in fibrinolytic parameters in male patients with diabetes mellitus (DM) and controls. DM patients were divided into three groups: patients without retinopathy, patients with simple retinopathy, and patients with proliferative retinopathy. Plasma levels of t-PA (tissue plasminogen activator) and t-PA-PAI-1 (plasminogen activator inhibitor-1) complex increased with increase in age, but those of PAI-1 (total and free) did not change in controls. On the other hand plasma levels of PAI-1 decreased with increase in age in DM patients. Plasma levels of t-PA, t-PA-PAI-1 complex, free and total PAI-1 increased with increase in body mass index in controls, but no significant changes were shown in these parameters in DM patients. When compared with controls, plasma levels of t-PA, t-PA-PAI-1 complex and PAI-1 were lower in DM patients. Plasma levels of UK (urokinase) and Lp(a) were higher in DM patients. ELT (euglobulin clot lysis time) was significantly shorter in DM patients than in controls. Patients without retinopathy showed increased fibrinolytic activities compared with those with retinopathy due to the increased levels of t-PA in plasma. These results seem to indicate that blood vessels release larger amounts of t-PA at the early stage of DM, then release being impaired at its advance stage. It is also suggested that the regulatory control mechanisms of fibrinolytic activity associated with mechanisms of fibrinolytic activity associated with change in age and body mass index are different between patients with DM and normal people. PMID- 8236168 TI - Antithrombotic activity of Desmin 370. Comparison with a high molecular weight dermatan sulfate. PMID- 8236169 TI - [The right patient in the right place]. PMID- 8236170 TI - [Thiazides in the treatment of hypertension--friends or enemies?]. PMID- 8236171 TI - [Quality control in the last round. Autopsy is still necessary!]. PMID- 8236172 TI - [Forensic analyses of causes of death and prolonged alcohol abuse]. PMID- 8236173 TI - [Causes of death among alcoholics]. AB - During the five year period 1987-91, 377 deaths among alcoholics were examined at the Gade Institute, Department of Forensic Medicine, University of Bergen. The material was dominated by male alcoholics. More than half the cases were regarded as natural deaths. Among these, the dominating cause of death was cardiovascular disease. The rest of the material, where death was attributed to non-natural causes, was dominated by lethal intoxications. A large number of the deaths were suicides. PMID- 8236174 TI - [Surgical treatment of fractures of the acetabulum]. AB - Since January 1988, 18 patients with acetabular fractures were operated on. Seven fractures were "both columns", six were posterior wall fractures, four were of the transverse/single column types, and one was unclassified. Four patients were operated by ileo-inguinal incision alone, three by combined ileo-inguinal and posterior Kocher-Langenbeck, ten by Kocher-Langenbeck, and one by triradiate incision. Early complications were 1 DVT and 1 infection (subsided after removal of the plate from crista iliaca). One patient with brain injury developed cerebral infarction, and one multi-traumatized patient died of lung embolus after transfer to another hospital. 11 patients are back at work, one has retired, and two are seeking new education after the injury. We conclude that operative treatment is indicated for these fractures, and that the results and rate of complications are acceptable. This treatment should be centralized. PMID- 8236175 TI - [Cerebellar infarctions]. AB - Infarction of the cerebellum comprises about 1.5 percent of all strokes. The symptoms are initially very similar to those of benign labyrinthitis, and the diagnosis is easy to miss. There are three major causes of clinical deterioration and death: Expansion of the infarct to the brainstem, swelling of the infarcted cerebellum with compression of the brainstem, and hydrocephalus. Surgical intervention may be lifesaving if hydrocephalus develops. Five patients admitted to our department are described. All described sudden vertigo or dizziness, which in three of them was accompanied by headache. All had ataxia. In four this was ipsilateral to the infarction and in one bilateral. Two had slight hemiparesis and plantar inversion. Two patients developed hydrocephalus. One was operated with external drainage. It is important to have this diagnosis in mind in all patients with acute labyrinthine symptoms. Acute CT scanning should be carried out if the patient becomes less conscious. PMID- 8236176 TI - [Recurrent abdominal pain. A prospective study of 68 children]. AB - We followed 68 children with recurrent abdominal pain in a prospective study including a standard investigation programme and abdominal ultrasound. We found an organic disease which explained the pain in eight patients (12%). Most children with recurrent abdominal pain can be dealt with by a general practitioner. In our opinion, further investigation, including abdominal ultrasound should only be carried out in the event of specific suspicion of organic disease. At follow-up 16-34 months later, 55 children (85%) had improved or were free of pain. We discuss the differential diagnoses and propose a plan of investigation for these children. PMID- 8236177 TI - [Villous glandular adenocarcinoma of the uterine cervix. A subtype with favourable prognosis?]. AB - The frequency of adenocarcinoma of the uterine cervix is increasing, and accounts for 10-20% of all cervical carcinomas. Adenocarcinoma usually affects women in their fifties and sixties. Mortality is high, with a five-year survival between 49 and 65%. Important prognostic factors are tumour grade, tumour diameter and clinical stage. The prognostic value of histologic subtyping is still under debate. Villoglandular (papillary) adenocarcinoma is a recently described form of well-differentiated adenocarcinoma in the cervix. This lesion apparently affects young women and carries an excellent prognosis. We describe five patients with villoglandular adenocarcinoma in the cervix, clinical stage IB and IIA. Mean age was 34.2 years and mean depth of cervical infiltration was 1.25 cm. Four patients were treated with radical hysterectomy and bilateral pelvic lymphadenectomy and one with simple hysterectomy. All five received postoperative chemotherapy. There is no evidence of recurrent tumour after a follow-up of 18 to 28 months. Cervical conization or simple hysterectomy is probably adequate treatment for this subtype of adenocarcinoma. PMID- 8236178 TI - [Transposition of submandibular ducts in the control of drooling]. AB - Drooling is a major problem in patients with cerebral palsy and various other neurological dysfunctions. Submandibular duct transposition for uncontrolled drooling is a technically straight forward operation. It has proved to be a safe procedure with low morbidity and a high rate of success. The authors review the literature, and present a material consisting of 21 patients from the University Hospital, Bergen, and Central Hospital, Rogaland. PMID- 8236179 TI - [Acute psychiatry]. AB - The author discusses psychological assessment and treatment of acute mental disorder in general practice. An initial assessment by telephone may provide the physician with important information about the nature of the problems and the patient's state of mind. Later, the immediate impression upon seeing the patient, and further observation during the consultation, are sources of important clinical information. The paper purports the view that psychological control over the situation is best achieved by means of psychological techniques adjusted to the specific behavioural disturbance in question. Drugs should only be used as one of several secondary options in the treatment of acute psychiatric disturbances with a non-organic etiology. PMID- 8236180 TI - [Accidental hypothermia]. AB - Accidental hypothermia is an important clinical condition in emergency and disaster medicine. It is usually classified as mild, moderate, severe, or extreme (body temperature below 18-20 degrees C; no recordable EEG activity). However, exposure time, trauma, serious illness, or the effects of drugs or alcohol may both attenuate and complicate the clinical course. This paper describes exposure mechanisms, the pathophysiologic processes, the body's thermo-regulating mechanism and diagnostic criteria. The author also discusses choice of treatment in the acute stages, during transportation and in hospital. The treatment should take into account not only the degree of hypothermia, but also exposure time, state of consciousness, and complicating factors such as trauma, drugs or alcohol. When hypothermia is associated with cardiac arrest, rewarming by extracorporal support is recommended. PMID- 8236181 TI - [Attitude of the public to autopsy. A questionnaire study]. AB - A high autopsy rate is a requirement for confirming or correcting clinical diagnoses and for providing opportunities for medical education on pre- and postgraduate level. A telephone survey was conducted to obtain information about the opinion of the Norwegian public on this matter. A randomised sample of 1,050 persons over 15 years of age were asked whether they would consent to autopsy if a close family member died in a hospital. Of the 954 (91%) who agreed to answer, 86% (95% CI 84-89) would consent, while 14% (95% CI 11-16) were unsure or would object. Demographical background variables did not significantly influence the response. When asked if they would give prior consent to an autopsy on themselves, should they die in a hospital, 84% (95% CI 82-87) said they would agree, while 16% (95% CI 13-18) were unsure or would object. Age between 31-50 years, male sex and married civil status significantly increased the likelihood of a positive attitude towards autopsy. The survey revealed a generally favourable attitude in the population towards autopsy. This positive confidence has to be maintained through careful talks with relatives about autopsy, and arrangements for relatives to be informed of the result of an autopsy. PMID- 8236182 TI - [Are thiazides diabetogenic?]. AB - Thiazide treatment is a first-line choice for hypersensitive patients: Thiazides have a flat dose-response as regards blood pressure, but induce biochemical changes only at higher doses. Given in low doses, which careful monitoring of potassium, thiazides provide adequate control of blood pressure without having any substantial impact on blood glucose. In high doses, thiazides may induce three to six new cases of impaired glucose tolerance per 1,000 patient years. The thiazide-induced changes in blood glucose can be prevented by giving lower doses and correcting hypokalaemia, and can be reversed by stopping the drug. In most diabetics, low doses of thiazides do not seem to diminish control of blood glucose. The well-documented effect of thiazides in reducing cardiovascular morbidity and mortality, their safety and their low price make them an excellent first-line choice for treatment of hypertension. PMID- 8236183 TI - [Environmental health promotion in municipalities--resources, organization and activities]. AB - The Municipal Health Services Act with amendments from 1988 defines environmental health promotion activities directed at physical, chemical, biological and social factors as mandatory for the local Norwegian health authorities. In addition, the municipal health services are responsible for health surveillance and for initiating cross-sectorial preventive measures. In 1991, we undertook a national survey among the Norwegian municipal health services to monitor planning activities, manpower resources, cross-sectorial cooperation, and project-oriented activities within the field of environmental health promotion. Less than one third of the municipalities employed technically trained hygienic personnel. However, three of four municipalities had carried out some environmental health promotion projects within the last two years. The following factors were all independently related to level of environmental health promotion activity: the availability of technical assistance, the level of cross-sectorial cooperation and the size of the population in the municipality. We conclude that this area of health promotion should be improved by better planning, a higher level of technical hygienic competence within the municipal health services, more inter sectorial cooperation and greater emphasis on visible projects of limited duration. PMID- 8236184 TI - [Intruders or innovators. More women physicians?]. AB - The article discusses the impact of an increasing proportion of women medical doctors in Norway. Will a changing gender composition affect the medical culture as such and, in the event, how? The theoretical point of departure is the social construction of gender. One argument is that gender relations tend to be reconstructed through gendered processes, many of them leading to new forms of gendered divisions of labour among doctors. Women doctors may be considered as the same as, or as different from their male colleagues. Such a distinction is often blurred, but is nevertheless operative. Medicine as a system of production of scientific knowledge is itself assumed to be primarily a product of male thoughts and ideas. Conventional forms and procedures may be challenged by its new members. Medical work as such runs the risk of being trivialized. To both understand and explain current developmental tendencies and contradictions, we need to identify gendered dynamics. There are no easy political solutions, however, to the "gender question" in medicine. PMID- 8236185 TI - [Tobacco death--the tuberculosis of our time]. PMID- 8236186 TI - [Can diabetes be cured by islet transplantation?]. PMID- 8236187 TI - [Blood glucose level means everything]. PMID- 8236188 TI - [We can prevent pain]. PMID- 8236189 TI - [Von Willebrand's disease--gene, molecule, clinical findings, treatment]. AB - In 1926, Erich von Willebrand described a bleeding disorder which, because of its autosomal inheritance and the tendency for spontaneous mucocutaneous bleeding, he called hereditary pseudohaemophilia. As a tribute the disease was named after him. However, its etiology and pathogenesis remained obscure until 1971, when Zimmermann and co-workers discovered that the blood of von Willebrand patients contained a lower than normal level of an antigen, the von Willebrand factor, which was associated with coagulation factor VIII. This paper briefly reviews the main points of what is known today about von Willebrand's disease: subtypes, gene, von Willebrand factor molecular/biological function relationships, prevalence, clinical expression, diagnosis and treatment. PMID- 8236190 TI - [Hodgkin's disease or lymphogranulomatosis. New views on prognosis, complicating conditions and complications after treatment]. AB - Between the years 1968-85, 1,177 patients started treatment for Hodgkins disease at the Norwegian Radium Hospital. In this unselected material from Norway the age distribution was bimodal with the highest incidence between 20-40 years of age. Survival is dependent on age, stage and histology, and was better between the years 1980-85 as compared with the period 1974-79. During the last ten years, treatment has been decentralized in Norway, and all centres cooperate by using the same protocol for staging and treatment. The article summarizes the results of the treatment, and related complications. PMID- 8236191 TI - [New cancer disease after treatment of Hodgkin's disease]. AB - The risk of a second cancer was assessed in 1,152 patients with Hodgkins disease who were treated at the Norwegian Radium Hospital from 1968-85. 68 patients developed a second cancer more than one year after the diagnosis of Hodgkins disease. These included nine acute non-lymphocytic leukemias, eight non-Hodgkins lymphomas and 51 solid tumours, including 11 lung cancers. The overall relative risk (observed/expected ratios) of developing a second cancer was 1.86. After 18 years the cumulative risk of developing a second cancer was 14.4% +/- 2.9%, of which 11.2% +/- 2.6% referred to solid tumours. The cumulative risk of leukemia appeared to reach a plateau level of 1.5% after 12 years while the risk of non Hodgkins lymphomas and lung cancer continued to rise with time to 2.1% and 3.3% respectively after 18 years. The risk of developing leukemia increased after treatment with alkylating agents and Procarbazine. The risk of non-Hodgkins lymphoma was not related to any specific type of therapy. Excess lung cancer risk was noted in patients treated with radiotherapy, and the cancers appeared within the treated areas. PMID- 8236192 TI - [Granulomatous mastitis in a patient treated with prednisone]. AB - A 36 year old woman, mother of a two year old child developed, in the course of one night, a tender mass in the upper medial quadrant of the left mamma. Treatment with antibiotics had no effect, and after a week the patient was admitted to hospital for drainage of the abscess and further examination. She had then developed reactive arthritis. Histological examination of a specimen from the mamma revealed lobular granulomatous mastitis. This connection has not been described before in the literature. Further examination showed no signs of infectious disease or sarcoidosis. Surgical drainage had only a minor effect on the breast-mass. The patient was treated with prednisone for six months, and after one year of observation the mass has disappeared, but the arthralgias persists. PMID- 8236193 TI - [Treatment of menorrhagia with tranexamic acid. A clinical trial of an underestimated problem in general practice]. AB - 44 women with heavy menstrual bleeding participated in a six months randomised, double blind, cross over clinical trial. The objective was to use simple methods to study whether treatment of menorrhagia with tranexamic acid has an effect that can be detected by the women themselves. The effect was evaluated by means of diaries and a visual analog scale. The bleeding during each period of treatment was compared with the bleeding during untreated periods. Bleeding during the different periods of treatment was also compared directly. The results indicate that tranexamic acid was more effective than placebo. This difference was not disclosed, however, in the laboratory data. After the study, 75% of the women wished to continue treatment with tranexamic acid. PMID- 8236194 TI - [Cardiac manifestations of Borrelia burgdorferi infection (Lyme-borreliosis)]. AB - Lyme borreliosis, caused by the tick-borne spirochete Borrelia burgdorferi, has been found to cause a variety of clinical syndromes including cardiomyopathy, dermatopathy, neuropathy, and arthropathy. Lyme carditis was originally described as a mild self-limited carditis, primarily involving the conduction system. However, recent reports suggest that cardiac involvement may be more serious than previously suspected, and may cause heart failure and probably congestive cardiomyopathy. PMID- 8236195 TI - [Body language in live communication]. AB - The core of every consultation is the patient-doctor interaction connected to the patient's problem. The physician's perception of this problem is based on an interpretation of words and of bodily signs. In Western medicine these two modes of expression are divided and separated from the relevant biographical connections. Taking case histories and clinical examination, defined and practised in accordance with the current epistemological traditions of science, unacceptably limit an understanding of human health or sickness. Some historical and theoretical traces are followed for the purpose of promoting a change in medical understanding. PMID- 8236196 TI - [New views on pathogenesis and treatment of pain caused by nerve injury]. AB - The article focuses on the pathophysiology of pain as a result of injury to the peripheral or central nervous system. The nervous system is dynamic, and nerve injury leads to long-lasting changes in the nociceptive neurons. Plasticity in the neuronal systems regulating nociception is important in explaining clinical features associated with nerve injury pain. To treat nerve injury pain more effectively, measures should be taken to both prevent and reverse the functional neuronal changes and plasticity caused by nerve injury. PMID- 8236197 TI - [Pain regulation and plasticity]. AB - The pathophysiology of chronic pain is not well understood. To try to explain why pain may become long-lasting or chronic, it is necessary to understand the plasticity of the neuronal circuits involved in nociception. It seems that mechanisms of learning and memory are involved in this process. Interactions between peptide and nonpeptide neurotransmitters may be important. Other significant factors appear to be changes in gene induction and transcription, and cell injury and destruction as a result of excitation. Through these mechanisms, painful states may induce changes in the nervous system which lead to greater pain, making adequate prevention and treatment of nociceptive pain essential. PMID- 8236198 TI - [Diagnosis of hemoglobinopathies and thalassemias]. AB - During recents the number of immigrants to Norway from Africa and South East Asia has risen considerably. These persons come from countries with a high prevalence of haemoglobinopathies. Most of the immigrants with haemoglobinopathies are silent carriers, but some have a serious disease or have offspring with serious disease. This situation calls for increasing awareness and knowledge of haemoglobinopathies, especially the thalassemias and sickle cell trait. Genetic counselling is particularly important, since the majority of these immigrants marry within small ethnic groups. This paper gives a survey of the most frequent haemoglobinopathies likely to be encountered in Norway. PMID- 8236199 TI - [Lipid-lowering medication--indications and possible hazards. Report from a hearing]. AB - Drug treatment of hyperlipidaemia should be founded on scientific evidence. A hearing was arranged to arrive at practical measures based on reported clinical trials. Several studies during recent years have shown regression of coronary atheromatosis, and some people would say the effect is striking. Studies using clinical endpoints, like morbidity and mortality, have not yielded equally convincing results, but it should be noted that the results of studies with more than 10% reduction in total serum cholesterol levels are not yet available. So far, the trials have been too small to judge effects on total mortality. It is uncertain whether the treatment actually has untoward effects on the incidence of violent deaths and neoplasms. An upper limit for drug treatment of about 8 mmol/l was proposed during the hearing. The limit can be lowered to 7 if established coronary disease is present. For women without coronary disease the limit should be higher than that for men. All in all, this implies an increase in the total number of individuals treated compared with present practice. It was agreed that at present, high risk individuals are undertreated. The new levels have been set partly on the basis of economic considerations. PMID- 8236200 TI - [Meningococcal project Telemark. Experiences after 5 years with contact tracing and eradication of the pathogenic bacteria in near contacts of the patients with meningococcal disease]. AB - Since 1987 we have analysed throat samples from 1,086 healthy contacts of 32 patients with meningococcal disease. The disease-causing strain was found in contacts of 17 out of the 32 patients. 161 (18%) of the contacts carried meningococci, and 30 (3%) of them were carriers of the disease-causing strain as determined by DNA fingerprinting. The carrier strain was eradicated in 29 of these 30 contacts by treatment with rifampicin. No secondary case of meningococcal disease was observed. During the four-year period 1984-87, there were 39 confirmed cases of meningococcal disease, including 12 verified and four suspected secondary cases of meningococcal disease. Therefore identification and eradication of the disease-causing strain seems to prevent secondary cases. A change in the Norwegian recommendations for preventing secondary cases of meningococcal disease should be discussed. PMID- 8236201 TI - [Role of the basic group in health insurance. Experiences with 70 cases in the municipality of Verdal]. AB - The role and conclusions of the discussions in the basic group in the municipality of Verdal have been evaluated. 70 patients were referred to the group during a three year period. The large majority of the patients suffered from minor mental disease or musculoskeletal disease. 63% of the patients and 69% of their own doctors attended the discussion. Follow-up, first after one year and later after two to four years showed a fair degree of reliability as regards the accuracy of the group's conclusions. The basic group is an important collaborator for the Norwegian National Insurance Scheme and should be consulted at an early stage in many cases evaluated for rehabilitation benefits or disability pension. PMID- 8236202 TI - [Physicians' reports about the mechanisms of whiplash injuries]. PMID- 8236203 TI - [Iron deficiency in teenagers]. PMID- 8236204 TI - [Problem-based teaching]. PMID- 8236205 TI - [The Nobel Prize winners' days 1993]. PMID- 8236206 TI - [Sense and concentration--about words and rules]. PMID- 8236207 TI - [Academic or practical social medicine?]. PMID- 8236208 TI - [Heart transplantation in terminal heart failure. Still a therapeutic challenge after 10 years]. PMID- 8236209 TI - [Organ transplantation--still uncovered needs]. PMID- 8236210 TI - [Is multiple personality a mental disorder?]. PMID- 8236211 TI - [Myths and expectations. Consequences of increased number of women physicians]. PMID- 8236212 TI - [Are sleeping pills dangerous to the elderly?]. PMID- 8236213 TI - [Use of hypnotics by elderly patients admitted for rehabilitation]. AB - 60 persons (13 men, 47 women, median age 80 years) who were admitted to a nursing home for rehabilitation were studied with respect to sleep-related problems and consumption of hypnotic drugs. 25 patients (42%) used hypnotics regularly. The median time of drug use was 72 months. More users than non-users reported difficulty in falling asleep (OR 3.1) and were less likely to feel rested in the morning (OR 4.4). There was no difference between users and non-users with respect to sex, age, living condition, use of analgesics or drugs that can influence sleep. In general, drug consumption was significantly higher among users of hypnotics than among non-users, and more of the users drank coffee or tea in the afternoon or evening. Ten of the users did not want to discontinue their drug consumption. PMID- 8236214 TI - [Multiple personality--a phenomenon also in Norway?]. AB - Dissociative disorders are little known and seldom referred to in the Norwegian medical literature. Our paper describes signs and criteria for multiple personality and dissociative syndromes, and reviews the recent literature. Three cases are briefly presented. PMID- 8236215 TI - [Analysis of the Duchenne muscular dystrophy gene with PCR analysis in paraffin embedded tissue. A new diagnostic possibility]. AB - Analysis of DNA from archival, paraffin-embedded muscle tissue allowed tracing of the mutated dystrophin gene in two families with Duchenne muscular dystrophy. There were no living patients in these families. In one family this diagnosis contributed to the birth of a normal male, in the other family to the demonstration of carrier status. These analyses, based on the polymerase chain reaction, are relatively rapid and simple, and lend increased value to old tissue samples stored in pathology departments. PMID- 8236216 TI - [DNA diagnosis of fragile X syndrome in a family. A new type of heredity--dynamic mutations]. AB - In typical cases, the diagnosis of fragile X syndrome can be made clinically, but has so far been based on cytogenetic detection of a fragile site on the X chromosome at Xq27.3. Cytogenetic analyses are time-consuming, and false negative results have been a problem. Following the successful molecular cloning of the gene causing the fragile X syndrome, a novel genetic principle was discovered: the "dynamic" DNA mutation. DNA analyses have now the power to distinguish between normal copies of the gene, carrier mutations and fragile X mutations. We have performed both cytogenetic and DNA analyses in an extended fragile X family. DNA fragment length variations in the gene were identified by Southern blot analysis, and the results were used diagnostically in the family. Instability of the mutation (dynamic mutation) occurred between generations and within individuals. DNA analysis, as performed here, improves diagnostic accuracy and genetic counselling in fragile X families. PMID- 8236217 TI - [Diagnostic aspects of subacute osteomyelitis in children and adolescents. Clinical and radiographic resemblance to primary malignant tumors]. AB - During a six-year period, 17 patients younger than 20 years of age, with a final diagnosis of subacute osteomyelitis, were admitted to the Norwegian Radium Hospital because of an initial suspicion of primary malignant bone tumour. The most common localizations were the metaphyses of long bones (eight patients) and the clavicle (four patients). Pain was the dominating symptom. Common radiological findings were localized osteolysis and/or sclerosis, cortical bone destruction, periosteal reaction and an adjacent, often palpable soft tissue mass. Clinical signs of infection were generally absent, and a positive bacterial culture was obtained from the biopsy material in only one patient. Following extensive investigations, a malignant bone tumour (especially Ewing's sarcoma) remained a differential diagnosis, and open biopsy was indicated in all cases. The patient material illustrates the difficulty in distinguishing between subacute osteomyelitis and malignant bone tumours, and it is stressed that diagnostic investigations for this type of patients should be performed in an oncological centre with experience of bone tumours. PMID- 8236218 TI - [Acute abdomen among children and adolescents. A retrospective study of 470 children and adolescents with acute abdominal pain]. AB - The causes of acute abdominal pain among children admitted to a surgical department were few and the fraction that needed surgical treatment was low (37%). The surgical intervention rate was age-dependent, rising from 11.4% (zero to three years of age) to 48.9% (12-15 years of age). The increase in surgical intervention rate was due to increasing incidence of acute appendicitis while the incidence of intestinal obstruction was unchanged during childhood. No child below the age of four had appendicitis, and the rate of perforated appendix among children seven years and younger (41.7%) was significantly higher than among children eight years and older (20.4%). For acute appendicitis, the surgeons' diagnostic accuracy was 77.9% and there was no significant difference between complications after appendectomy for appendicitis and complications after negative laparotomy. The diagnostic value of biochemical measurements was limited. However, the combined evaluation of C-reactive protein measurements and leucocyte counts possibly supports further observation rather than immediate operation. PMID- 8236219 TI - [Clinical knowledge--facts or something more? Some theoretical views]. AB - Origins, contents and validation procedures of medical knowledge are rarely discussed in medicine. Clinical knowledge is the core of medicine. Analysis of the procedures and strategies for seeking clinical knowledge, and the content of this knowledge, may enlighten the development of medical theory as well as constitute a basis for quality assurance of medical practice. The article presents epistemological challenges related to analysis of clinical knowledge, and delivers some theoretical frames of reference (postmodernism, pragmatism, practice theory and feminism) with a potential to provide a broader understanding of the medical field--acknowledged as living knowledge produced through human interaction. PMID- 8236220 TI - [Molecular genetic causes of familial hypercholesterolemia]. AB - Familial hypercholesterolemia (FH) is characterized by autosomal dominantly inherited hypercholesterolemia, xanthomas and premature coronary heart disease. The disease is caused by defective cell surface low density lipoprotein (LDL) receptors. This paper reviews the molecular defects underlying defective LDL receptors. Studies of the molecular genetics of FH have provided important information on the structure-function relationship of the LDL receptor. This information may explain the considerable variation in levels of serum cholesterol among FH subjects, and why FH subjects respond differently to lipid lowering drugs. The diagnostic and therapeutic implications of the molecular findings are discussed. PMID- 8236221 TI - [Cadaver organ donation in Norway 1989-92]. AB - The annual number of cadaveric organ donors in Norway increased from an average of 12.4 per million inhabitants per year in the period 1985-88 to 13.1 in the period 1989-92. An increase to 20 organ donors per million per year is required to meet the anticipated need for organs. The donation rates differ greatly between health regions. Possible strategies to increase the organ donation rate are discussed. PMID- 8236222 TI - [Establishing smoking habits among adolescents. A study of 9th-grade students]. AB - A cross-sectional study of 661 9th grade pupils in Bergen municipality was conducted in November 1992. The pupils completed an anonymous questionnaire on their smoking habits. 25% of the girls and 16% of the boys reported smoking daily. Among the daily smokers, the average daily consumption of cigarettes was seven for both sexes. Most of the experimenting with cigarettes took place from the 6th to the 8th grade. Most youngsters tried their first cigarette when outdoors with friends. 27% of the regular smokers reported smoking at home, and 40% at school. In spite of legislation banning the sale of cigarettes to persons below the age of 16 years, 75% of the smokers bought the tobacco themselves. PMID- 8236223 TI - [Health work in Guatemala among returning refugees]. PMID- 8236224 TI - [Norwegian drug policy--socioeconomically unprofitable, hostile to patients and to research]. PMID- 8236225 TI - [Embryo research: ethics of technology and technique of ethics]. AB - A short survey is given of the aims and methods used in embryo research. Ethical aspects of research and possible applications lead to the conclusion that questions are raised that concern society as a whole. In the last decade the awareness in this respect has increased. Methodology of broad ethical judgement and decision making however is still in a stage of development. It is suggested to strive for: 1) clarification of fundamental views on values in (human) life, 2) answering the question: who decides about what, and 3) determination of decision making procedures. PMID- 8236226 TI - [Twins in cattle. Freemartin or not? Current aspects]. AB - Methods are described for the detection of freemartins. The first step is to measure the length of the vagina, followed, if necessary, by laboratory investigations--blood group, chromosome, or DNA typing. PMID- 8236227 TI - [Feline fibro-epithelial hyperplasia in 3 castrated tomcats following treatment with progestagens]. AB - This article describes three castrated tomcats that were treated with progestins. All three developed symptoms of fibroepithelial hyperplasia of the mammary gland, probably as a consequence of this treatment. This disease is commonly found in intact young female cats, pregnant cats, and progestin treated cats. Therapeutical consequences are dealt with and the importance of (histo)pathological investigation to 'Good Veterinary Practice' is pointed out. PMID- 8236228 TI - [Epidemiology for application in the Tropics]. PMID- 8236229 TI - [Udder infections]. PMID- 8236230 TI - [Rottweiler male dog. What is your diagnosis? Vitiligo/poliosis]. PMID- 8236231 TI - [Castration for behavior correction]. PMID- 8236232 TI - [Salmonella enteritidis--monitoring and prevention in reproduction poultry in 1992]. PMID- 8236233 TI - [One and one half centuries Royal Dutch Society for Veterinary Medicine, department Groningen/Drenthe]. PMID- 8236234 TI - HLA-DR and -DQ genotyping by PCR-SSO in Shanghai Chinese. AB - The polymorphism of HLA class II (HLA-DRB1, -DQA1 and -DQB1) were investigated in 89 Shanghai Han Chinese using polymerase-chain reaction amplification and oligonucleotide (PCR-SSO) typing. Of the 43 DRB1 alleles tested, 24 were observed. DRB1*0901 (16.85%), *0803 (9.55%), *1202 (9.55%) and *1501 (12.92%) were most frequent and account for the high frequencies of DR9, DR8, DR5 and DR2 specificities in this population. The alleles DRB1*0101, *0102 and *1001 had very low frequencies. Among the 8 DQA1 and 13 DQB1 alleles tested, only DQA1*0401 and DQB1*0402 were absent. DQA1*0301, and DQB1*0301 and *0303 were among the most common alleles of the two loci respectively. Unusual DRB1-DQA1-DQB1 haplotypes were observed and a DRB1 "new" allele was suggested. PMID- 8236235 TI - The HLA polymorphism of two distinctive South-American Indian tribes: the Kaingang and the Guarani. AB - The HLA-A, B, C, DR and DQ antigens of 240 Kaingang and 98 Guarani individuals have been characterized. The most frequent antigens found among the Kaingang are A31, 2, 24; B35, 51, 39, 48; Cw4, 7, 3, 1; DR8, 4, 2; DQ blank, 3. In the Guarani, they are A2, 28, 31; B40, 62, "53G"; Cw3, 4; DR2, 4, 8, 6; DQ3, blank. B " 53G" is an unusual antigen of the B5 cross-reactive group. DQ blank possibly corresponds to DQ4, not tested in this study. The reaction patterns of B35, B40 and DR4 indicate intra-tribal (of B35 and B40), and inter-tribal (DR4, B40 and B35) heterogeneity of these antigens. 408 Kaingang and 141 Guarani haplotypes were defined by segregation analysis. Of the commonest 10 Guarani and 9 Kaingang haplotypes, only one is shared by both tribes. Significant, positive linkage disequilibrium values for HLA-A,B; HLA-A,C; HLA-B,DR and most HLA-B,C antigen pairs were also different for the two populations. Genetic distance estimates between these two and another seven South-American Indian populations, and relative to the major human races (negroids, caucasoids, and mongoloids) reveal a comparatively high degree of divergence between the Kaingang and the Guarani, which is uncommon for Amerindian populations living close one to another. PMID- 8236236 TI - HLA class II DNA typing in two Brazilian populations. AB - Brazil constitutes a melting pot of populations arising from three major groups, including Amerindians, Africans, and Europeans predominantly from Portugal who were later supplemented by migrations from other European countries. Although every possible combination of racial mixture exists in Brazil, we have selected for this study two groups of subjects residing in Rio de Janeiro. A predominant White population, among whom some Amerindian admixture may exist, and a predominantly African population having little admixture from the other races. We have used the polymerase chain reaction (PCR) and hybridization with oligonucleotide probes to perform a complete typing of the HLA class II alleles. We report the allele frequencies for HLA-DRB1, DQA1, DQB1 and DPB1. We also report on the postulated DR-DQ haplotypes based on family studies and observations in homozygous B-cell lines. These results may serve as background for various types of clinical studies in Brazilian populations. PMID- 8236237 TI - A novel HLA-DPB1 allele (DPB1*4501) in a Dutch Caucasian healthy control. PMID- 8236238 TI - A novel HLA-DQB1 allele: evidence for gene conversion event promoted by chi-like sequence at DQB1 locus. PMID- 8236239 TI - Analysis of genetic variables in selective human IgG3 deficiency. PMID- 8236240 TI - New DP sequences: three DPA1 and one DPB1. PMID- 8236241 TI - Perioperative alterations in polymorphonuclear leukocyte function of gastrointestinal surgery. AB - To characterize the alteration of perioperative polymorphonuclear leukocytes (PMNs) function in surgical stress, we studied twenty six patients undergoing gastrointestinal surgery. Seventeen patients with thoracic esophageal cancer underwent total thoracic esophagectomy through a right thoracotomy (severe surgical stress group). Nine patients underwent cholecystectomy (slight surgical stress group). Phagocytic oxygen-dependent microbicidal activity in the esophagectomy patients significantly increased postoperatively, by measuring O2- production (35.3 +/- 7.0 nmol/10(6) cells/ml/20 min on postoperative day 1 vs. 28.6 +/- 6.2 preoperatively, p < 0.01) and luminol-dependent chemiluminescence (99.5 +/- 29.9 x 10(5) cpm/10(5) cells on postoperative day 3 vs. 67.5 +/- 12.8 preoperatively, p < 0.01). On the other hand, only a slight change was seen in the cholecystectomy patients. We conclude that the postoperative PMN function in terms of oxygen-dependent microbicidal activity significantly increases when the degree of surgical stress is sufficient. In order to gain insight into the mechanism of PMN activation, we specifically analyzed the expression of complement receptors. Up-regulation of complement receptors were seen in the esophagectomy patients, which parallels the activation of PMN microbicidal activities. It was suggested that phagocytic PMN function in severe surgical stress significantly increases postoperatively, in part, based on the upregulation of cell surface complement receptors. PMID- 8236242 TI - Acute hemodynamic changes after Fontan operation: an experimental study. AB - Seven adult mongrel dogs with an average body weight of 11.3 kg were subjected to experimental Fontan operation in order to evaluate acute hemodynamic changes at the Fontan circulation. The Fontan circulation was established by occlusion of the tricuspid valve with tightening a purse-string suture which was placed around the valve under inflow occlusion and was passed through the anterior right atrial wall, and by connection with a 10 mm Gore-Tex graft between right atrium and pulmonary artery. When the Fontan circulation was established, aortic pressure and cardiac output (CO) decreased from 94.9 +/- 22.0 mmHg to 41.9 +/- 6.0 mmHg and 0.907 +/- 0.179 liter/min to 0.259 +/- 0.072 liter/min, respectively. There were no significant changes in heart rate and systemic vascular resistance between normal and the Fontan circulations. On the other hand, pulmonary vascular resistance (PVR) increased from 11.6 +/- 7.3 units in the normal circulation to 21.8 +/- 13.0 units in the Fontan circulation. From this study it was concluded that main changes in hemodynamic parameters immediately after establishing the Fontan circulation were marked decrease in systemic arterial pressure and CO, and an increase in PVR. Therefore, one should consider a possibility of an increase in the PVR to the critical level when the Fontan operation was applied to patients with pulmonary hypertension. PMID- 8236243 TI - Stimulation of pulmonary intravascular macrophages increases microvascular permeability in awake sheep. AB - The purpose of this study is to determine if stimulation of pulmonary intravascular macrophages (PIMs) increase microvascular permeability in sheep. We infused latex microbeads, 1 micron in diameter, for 1 hr continuously and analysed lung hemodynamic and lymph-dynamic changes. More than 70% of latex microbeads in the lung were assigned to PIMs, and caught in their phagosomes as determined by morphological examination. This implies that infused latex microbeads predominantly stimulate PIMs. Pulmonary arterial pressure increased during the infusion period, and returned to baseline after the infusion period. Lung lymph flow increased and remained high while the lymph to plasma protein ratio ultimately increased above baseline. This implies that infusion of latex microbeads increases pulmonary microvascular permeability. The increase in lung lymph protein clearance was blocked completely by pretreatment with indomethacin, but not with a thromboxane synthetase inhibitor (OKY-046). These data indicate that the increase in microvascular permeability is mediated by an arachidonic acid cyclooxygenase metabolites but not by thromboxane. We conclude that PIMs can act as an initiator to increase pulmonary microvascular permeability by releasing arachidonic acid cyclooxygenase metabolites through their stimulation with latex beads. PMID- 8236244 TI - The effect of topical tretinoin on the photodamaged skin of the Japanese. AB - Fifteen middle aged or elderly patients with chronic solar damage of the skin, eight patients with melasma and three patients with xeroderma pigmentosum were treated with topical tretinoin for 6 months. There was a significant improvement in fine surface lines in periorbital region, but no significant improvement was observed in deep furrows. No significant change was induced in melasma despite the improvement in smoothness of the skin surface. Global improvement was also seen in one patient with xeroderma pigmentosum. With regard to the functions of the stratum corneum that was assessed on the flexor surface of the forearms, values of water content as well as transepidermal water loss were found to increase one month after start of the application of tretinoin cream. On the other hand, there was no significant change in the amino acid content of the stratum corneum when measured after 4 months of the treatment. It is concluded that tretinoin cream is capable of partly reversing fine surface lines in photodamaged facial skin of the Japanese. However the irritation induced by 0.1% tretinoin cream was unexpectedly severe in the Japanese as compared to that reported in Caucasians. PMID- 8236245 TI - Reversal of glucose insensitivity of pancreatic B-cells due to prolonged exposure to high glucose in culture: effect of nicotinamide on pancreatic B-cells. AB - Prolonged in vitro exposure to high glucose has been shown to induce a decrease in pancreatic B-cell sensitivity to glucose stimulation. This experiment was designed to study whether nicotinamide affects the B-cells' response to regulatory stimuli of glucose after prolonged culture with high glucose. Neonatal pig pancreatic islet-like cell clusters (ICCs) formed from single cells were embedded in agarose-gel. Some of them were maintained in RPMI 1640 containing a high glucose concentration (16.7 mM) and 10% fetal bovine serum (FBS) with or without nicotinamide. Then, the embedded-ICCs were stimulated by glucose at concentrations of 5.5 mM and 16.7 mM. After the prolonged culture with high glucose, the ICCs showed no response to acute glucose stimulation. There was a significant increase in glucose-mediated insulin secretion when the ICCs were maintained with the medium containing nicotinamide. We conclude that nicotinamide could protect B-cell desensitization to glucose after prolonged exposure to high glucose. PMID- 8236246 TI - Simultaneous determination of organotin compounds in fish and shellfish by gas chromatography with a flame photometric detector. AB - In the present study the levels of the organotin compounds were determined in fish and shellfish which were purchased from retail markets in Niigata, Japan. The concentrations of dibutyltin (DBT), tributyltin (TBT), diphenyltin (DPT) and triphenyltin (TPT) compounds were separately determined by FPD-gas chromatography. Eight species (yellowtail, tuna, cuttlefish, olive flounder, northern shrimp, cinnamon flounder, Japanese sea bass and oyster) were studied and five samples of each species were analyzed in each season, which meant 40 samples in one season, and 160 in total. Among the compounds of interest, DBT, TBT and TPT were detected. The highest concentrations detected were 0.674 microgram/g for DBT, 0.669 microgram/g for TBT and 0.186 microgram/g for TPT. DBT was detected as much as TBT, however, its concentration was not correlated with that of TBT. Seasonal changes of the mean value of the concentration of DBT and TBT were observed. In some species, such as yellowtail and Japanese sea bass, combined contaminations of these organotin compounds were also evident. This study is the first step to elucidate possible health hazards by environmental pollution of organotin compounds on human beings. PMID- 8236247 TI - Plasma GH response to the sequential 3 day administrations of GHRH followed by arginine infusion in patients with idiopathic GH deficiency and normal short children. AB - To study the site of lesions in idiopathic growth hormone (GH) deficiency (IGHD), growth hormone releasing hormone (GHRH) was administered sequentially for 3 days to 19 patients with IGHD, 3 patients with GH deficiency (GHD) secondary to hypothalamic tumors, and 7 normal short children (NSC). GHRH (100 micrograms) was injected as a bolus on days 1 and 3, and was infused over 60 min on day 2. Of 19 patients with IGHD, 6 showed an improved GH response (group A), 5 a decreased response (group B) and the remaining 8 an unchanged response (group C) to sequential administration of GHRH. The response was unchanged in patients with secondary GHD or NSC. There was no significant correlation between the patterns of GH response and the findings on pituitary MR images or the delivery state at birth in IGHD patients. Ten patients with IGHD (4 of group A; 3 each of groups B & C) and 2 NSC showed much greater GH responses to arginine (0.5 g/kg i.v. for 30 min) injected with preceding GHRH than to arginine injected without preceding GHRH. These results indicate that hypothalamic lesions were primarily responsible for GH deficiency in about 60% of the patients with IGHD (groups A and B), and group C might have more severe hypothalamo-pituitary damages than the other groups. Hypothalamic somatostatin neurons seems to be functioning to a degree even in severe IGHD patients. PMID- 8236248 TI - Relative formation of dichloroacetate and trichloroacetate from trichloroethylene in male B6C3F1 mice. AB - The hepatocarcinogenicity of trichloroethylene (TRI) has been attributed to the metabolite trichloroacetate (TCA). However, mice also form dichloroacetate (DCA) and trichloroethanol (TCE) as metabolites of TRI. TCA and DCA have both been shown to induce hepatic tumors in mice. This study was undertaken to measure the kinetics of TCA and DCA formation in the B6C3F1 mouse using doses of TRI ranging from 0.38 to 15 mmol/kg and TCA at doses of 0.03 to 0.61 mmol/kg. The formation and elimination of TCA and DCA have been found to be nonlinear with the dose of TRI. Quantifiable levels of DCA were found in blood with doses above 0.76 mmol/kg TRI. The peak concentration of DCA did not show an appreciable change with an increased dose; however, the area under the curve (AUC) increased linearly with respect to the dose of TRI. Both peak concentration and AUC of TCA and TCE increased in a linear manner to a dose of 3.8 mmol/kg. The kinetics of TCA elimination following doses of TCA were similar to those found for TCA following doses of TRI. A significant dose-dependent partitioning of TCA into blood over liver was found at the higher doses of TRI and TCA investigated. Binding of TCA to plasma constituents accounted for this distributional pattern. Prior work has documented that DCA can be formed from TCA. However, the AUC for DCA following TRI exceeds that predicted from the formation of TCA from TRI. Additional pathways would, therefore, appear to account for the formation of DCA. Results from this investigation suggest that sufficient concentrations of DCA appear to be formed and may contribute to the hepatocarcinogenicity of TRI. PMID- 8236249 TI - Differences in the neuroexcitatory actions of pyrethroid insecticides and sodium channel-specific neurotoxins in rat and trout brain synaptosomes. AB - The effects of pyrethroid insecticides and other sodium channel-specific neurotoxins on synaptosomal membrane potential were investigated in rat and trout brain synaptosomes using the membrane-permeant lipophilic cation [3H]tetraphenylphosphonium (TPP+). Concentration-dependent and tetrodotoxin sensitive decreases in TPP+ accumulation, indicative of membrane depolarization, were produced by veratridine, aconitine, scorpion (Leiurus quinquestriatus) venom, and type I and type II pyrethroids, in both species. Veratridine, aconitine, and Leiurus venom were more potent and efficacious membrane depolarizing agents in rat synaptosomes than in trout synaptosomes. Type II (deltamethrin, cypermethrin) pyrethroids produced similar depolarizing responses in rat and trout synaptosomes; however, the 1R-cis-alpha R isomer of deltamethrin, which had no effect on membrane potential in rat synaptosomes, depolarized trout synaptosomes. This isomer of deltamethrin was also shown to produce toxicity in trout, but not in rats. The type I pyrethroids, permethrin and NRDC 157, exhibited significantly greater intrinsic activity in trout brain synaptosomes, producing maximal membrane depolarizations that were three times greater than those observed in rat brain synaptosomes. These results provide evidence of species-specific differences in the membrane-depolarizing properties of pyrethroid insecticides and sodium channel-specific neurotoxins. They also suggest that some of the neurotoxin binding domains of the voltage-sensitive sodium channel in trout brain differ from those in mammalian brain. The hypersensitivity of fish to the neurotoxic actions of pyrethroid insecticides may be related to these differences. PMID- 8236250 TI - A physiologically based pharmacokinetic model for nasal uptake and metabolism of nonreactive vapors. AB - A PB-PK model has been developed for nasal nonreactive vapor uptake in the F344 rat which incorporates nasal enzyme distribution as well as nasal airflow patterns. Nasal tissue is separated into respiratory and olfactory mucosal areas with each area containing mucus, epithelial, and submucosal compartments. Metabolic activities are distributed among the compartments in accordance with published histochemical data, and intercompartmental transfer rate constants are based on molecular diffusivity. Two airflow paths are assumed: a lateral/ventral path which passes over only respiratory mucosa and a dorsal medial path which passes over both respiratory and olfactory mucosa. This model was fit to F344 rat nasal uptake data obtained for five vapors: acetone (an unmetabolized vapor), isoamyl alcohol (alcohol dehydrogenase substrate), ethyl acetate (carboxylesterase substrate), and o-xylene and bromobenzene (mixed function oxidase substrates). These vapors cover a wide span of uptake values ranging from a fractional uptake of less than 0.01 for xylene to 0.80 for isoamyl alcohol. The model accurately predicted the fractional uptake of these vapors, the maximal deviation between actual and predicted values being 0.04. The best fit for the data was obtained assuming: (1) an asymmetric blood flow pattern (with the majority of the blood flow being apportioned to the respiratory mucosa), and (2) only a small fraction of the total airflow (8%) following the dorsal medial pathway and penetrating to the olfactory mucosa. Since only a small fraction of the inspired air passes over the olfactory mucosa, a site with high metabolic activity, an inspiratory flow rate dependence on inspired vapor metabolism results. At low flows, olfactory metabolism is limited by low vapor delivery to that site, an effect that does not occur at high flows. This highlights the potential importance of regional nasal airflow and enzymatic activity patterns in influencing uptake and metabolism of inspired vapors and indicates the need for physiologically relevant models incorporating these factors. PMID- 8236251 TI - Effects of in utero methylmercury exposure on a spatial delayed alternation task in monkeys. AB - Adult female monkeys (Macaca fascicularis) were exposed to 0, 50, 70, or 90 micrograms/kg/day of methylmercury prior to and throughout pregnancy and produced 11, 9, 2, and 2 infants, respectively. At birth, blood mercury levels of treated infants ranged from 1.04 to 2.46 ppm. At approximately 7 to 9 years of age, the monkeys were trained by successive approximation to respond on a lit button for a small amount of apple juice. The monkeys were then trained on a 0.1-sec spatial delayed alternation task to a specified criterion of performance. This was followed by 10 sessions each of fixed delay times of 0.5, 1, 3, 5, and 10 sec, followed by 20 sessions containing variable delay times of 0.1 to 15 sec. Data from all treated monkeys were combined. There were no differences between treated and control monkeys in initial button training or number of sessions to reach criterion on 0.1-sec delay procedure. On the fixed delay sessions, the treated monkeys had significantly more correct trials, and fewer incorrect responses, perseverative responses, and delay responses than controls. There were no differences between the treated and control monkeys on performance on the variable delay schedule. Results from this study indicate that in utero methylmercury exposure did not adversely affect the spatial memory of adult monkeys when tested on a delayed alternation task and may have facilitated performance on this task. PMID- 8236252 TI - Species differences in nucleotide pool levels of 2',3'-dideoxycytidine: a possible explanation for species-specific toxicity. AB - The nucleoside analogue, 2',3'-dideoxycytidine (ddC), a potent inhibitor of human immunodeficiency virus reverse transcriptase (in its anabolized triphosphorylated form), mediates virologic and immunologic improvements in AIDS patients. Clinical studies using ddC have shown various ddC-related toxicities, the most pronounced being a dose-limiting peripheral neuropathy. The dose responsiveness and manifestation of the ddC-related neuropathy vary among species, with greatest sensitivity in human > monkey > rabbit whereas mice and rats are insensitive to ddC-related neuropathy. This study has examined nucleotide pool sizes of ddCTP and its constituents (ddC, ddCMP, ddCDP) in cultured fibroblasts (human, rabbit, mouse) and freshly isolated peripheral lymphocytes (monkey, rabbit, rat, and mouse). Cells were treated with 10 microM [3H]ddC and nucleotide pool sizes analyzed by HPLC. The formation of nucleotide pools increased during the 24-hr assay period. Fibroblast pool formation of phosphorylated metabolites was significantly greater in human > rabbit > mouse. Lymphocytes demonstrated a similar pattern with monkey > rabbit > mouse = rat. Total ddC anabolite pools were also found to be significantly smaller (p < 0.05) in rodent lymphocytes than in those of rabbit or monkey, and rodent fibroblasts were smaller than those of human or rabbit (p < 0.05). These findings indicate that nucleoside phosphorylation and intracellular levels of phosphorylated metabolites may play an important role in determining species sensitivity and manifestation of ddC related toxicity. PMID- 8236253 TI - Species-specific cutaneous biotransformation of the pesticide propoxur during percutaneous absorption in vitro. AB - Propoxur (2-isopropoxyphenyl N-methylcarbamate) is a pesticide with a wide spectrum of applications, including use in agriculture and greenhouses. Percutaneous absorption and concurrent cutaneous metabolism of propoxur were studied in a two-compartment organ culture model. Nontoxic concentrations of [14C]propoxur were applied topically to skin discs from human, rabbit, and porcine origin. Permeation rates were comparable in human and rabbit skin, while pig skin was found to be twice as permeable. Furthermore, it was demonstrated that skin tissue of all three species had the capacity to metabolize propoxur. Hydrolysis of propoxur yielded 2-isopropoxyphenol (IPP), followed by phase II conjugation reactions. Interestingly, the type of IPP conjugation appeared to be species specific. In porcine skin cultures, glucuronides and sulfates were detected in equal amounts, whereas in human skin only sulfate conjugation was observed. For rabbit skin, glucuronidation was the major route of conjugation, with minor amounts of the sulfate conjugate and an unidentified metabolite. The percentage of propoxur metabolism in rabbit skin was not influenced by the dose in the range of 25-200 micrograms/cm2; in contrast, human skin metabolism was virtually saturated at 100 micrograms/cm2. PMID- 8236254 TI - Decreased expression of cytochrome P450 mRNAs and related steroid hydroxylation activities in hepatic hyperplastic nodules in male F344 rats. AB - To explore the mechanism(s) underlying the relationship between expression of cytochrome P450 (CYP) enzymes and growth of hepatic hyperplastic nodules, mRNA of CYP1A1, 1A2, 2B1, 2B2, 2C6, 2C11, and 3A1, and CYP enzyme-mediated hydroxylation of testosterone (T), 17 beta-estradiol (E), and corticosterone (C) were determined in a group of selected large nodules from livers of male F344 rats given the modified Solt-Farber resistance protocol and compared with control and surrounding tissue. Slot-blot analysis with specific oligonucleotides showed little or no mRNA of CYP2B1, 2B2, and 2C6 in nodules, surrounding tissue, and control liver without PB treatment. Constitutive expression of CYP1A2, 2C11, and 3A1 in nodules was from 2 to 10 times less in nodules than in control liver. BP treatment increased mRNA of CYP2B1, 2B2, 2C6, 2C11, and 3A1 in control and surrounding tissue substantially, but was 3 to 11 times less in the nodules than in the control liver. Messenger RNA of CYP1A1 was not detected in either nodules or control livers independent of PB treatment. HPLC determination demonstrated a general pattern of a decrease in CYP enzyme-mediated hydroxylation of T, E, and C in the nodules relative to the surrounding tissue and control liver. The differences between nodules and control liver were usually 3 to 10-fold for the 12 metabolites detected although smaller decreases were observed for a few metabolites in some nodules. PB increased the rate of hydroxylation for 11 of the 12 metabolites and resulted in 2 additional metabolites of both T and E in control liver and some nodules. However, the general pattern of lower hydroxylation activities in the nodules relative to the surrounding tissue remained the same as that in the non-PB group. These observations are consistent with the hypothesis that intracellular kinetics of endogenous CYP enzyme substrates may differ in some nodules, relative to the surrounding tissue, as a result of decreased expression of CYP genes. This difference in CYP expression pattern in turn may contribute to the selective growth and progression to cancer of certain nodules, as many endogenous CYP enzyme substrates are modulators of DNA and RNA synthesis and cellular growth and differentiation. PMID- 8236255 TI - Thymic atrophy caused by ethanol in a mouse model for binge drinking: involvement of endogenous glucocorticoids. AB - Consumption of large amounts of ethanol (EtOH) in a single drinking episode is common, but very little is known about the immunological effects of such occurrences. Exposure to EtOH for several days is immunosuppressive in rodent models, and a single dose of EtOH causes substantial increases in endogenous glucocorticoid levels which might have immunosuppressive effects. In the present study, the effects of a single dose of EtOH on the thymus and the role of endogenous glucocorticoids in these effects were examined in B6C3F1 female mice. A single dose of EtOH decreased thymus weight and cellularity, predominantly by elimination of CD4+CD8+ (immature) thymocytes. This occurred over a broad range of EtOH doses and was associated with behavioral effects (ranging from mild ataxia to unresponsiveness) similar to those noted in human binge drinkers. Several lines of evidence indicate that the effects of EtOH on the thymus are mediated by endogenous glucocorticoids: (1) corticosterone levels in EtOH-treated mice increased more than 10-fold and remained significantly elevated for up to 12 hr; (2) the most glucocorticoid-sensitive thymocytes (CD4+CD8+ cells) were preferentially depleted by EtOH; (3) before thymocyte depletion was evident, substantial DNA fragmentation occurred in the thymus as would be expected in the case of glucocorticoid-induced apoptosis; (4) the glucocorticoid antagonist, RU 486, blocked thymic atrophy and DNA fragmentation in EtOH-treated mice; (5) EtOH and its major metabolites at concentrations comparable to or greater than expected in vivo did not decrease thymocyte viability in 20-hr cultures, indicating that direct action of EtOH or its metabolites on thymocytes does not play an important role in EtOH-induced thymic atrophy. These results suggest that a single dose of EtOH induces thymic atrophy which is predominantly mediated by increased levels of endogenous glucocorticoids. The mouse model described here should be useful in evaluating other effects of binge drinking on the immune system, and the experimental strategy described should be applicable in investigating the role of endogenous glucocorticoids in thymic atrophy induced by other chemicals and drugs. PMID- 8236256 TI - Immunohistochemical localization of the cytochrome P450 isozymes LMC2 and LM4B (P4501A1) in 2,3,7,8-tetrachlorodibenzo-p-dioxin-treated zebrafish (Brachydanio rerio). AB - Dibenzo-p-dioxins are persistent pollutants in our environment that exert a variety of biological and toxic effects in various species, in particular developmental toxicity in fish. Using zebrafish (Brachydanio rerio) as a model system, we analyzed the expression and inducibility of a constitutive isozyme (LMC2) and a dioxin-inducible isozyme (LM4B or P4501A1) of cytochrome (cyt.) P450 in different organs by immunohistochemical techniques. In untreated zebrafish, LMC2 was constitutively expressed at high levels in the liver, kidney, skin, and oral mucosa, while moderate expression was detected in gills, pseudobranch, intestine, and ovaries. LM4B was either not expressed in these organs or was found at comparatively low levels. After treatment of zebrafish with 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD; total dose of 15 ng per fish), LM4B expression was strongly induced in livers and kidneys and less pronounced in gills and pseudobranches, whereas no or only marginal induction was seen in skin, intestine, oral mucosa, and ovaries. In addition, cyt. P450 catalyzed 7 ethoxyresorufin-O-deethylation and phenol UDP-glucuronosyltransferase activity were also found to be inducible in liver and kidney. Our results demonstrate that zebrafish respond to inducing stimuli of TCDD in a fashion similar to that observed with related compounds in other fish species, suggesting that zebrafish may be a useful experimental model for studying biological and toxic effects of TCDD and other environmental pollutants in fish. PMID- 8236257 TI - Mercury blockade of thiazide-sensitive NaCl cotransport in flounder urinary bladder. AB - The effects of HgCl2 on ion transport were investigated using isolated sheets of flounder urinary bladder, a model epithelium that is capable of electrically silent NaCl absorption and electrogenic K secretion. Exposure of the mucosal surface of the bladder to submicromolar doses of HgCl2 reduced K secretion, but the effect was not due to blockade of apical K channels. Rather, the effects of HgCl2 were virtually identical to those seen with experimental maneuvers that blocked the thiazide-sensitive NaCl cotransporter in the apical membrane, e.g., hydrochlorothiazide, Cl-free solutions, and Na-free solutions. Mucosal HgCl2 also blocked 22Na absorption, suggesting that the effect of the metal was mediated by blockade of NaCl entry. The effects of HgCl2 had a rapid onset and were readily reversed by washing, suggesting a noncovalent binding reaction. The abundance of polyanionic Hg complexes in salt solutions prompts the speculation that one of these may bind to the Cl-binding site on the cotransporter, thereby blocking it. The results provide the first evidence that the thiazide-sensitive NaCl cotransporter is a specific site of action for inorganic mercury. PMID- 8236258 TI - Comparison of the effects of losulazine and reserpine on central aminergic neurons. AB - The purpose of this study was to examine the effects of both acute and chronic administration of the peripheral sympatholytic antihypertensive agent losulazine on central dopaminergic, noradrenergic, and 5-hydroxytryptaminergic neurons in the rat. For comparison, the acute effects of reserpine were also examined. Acute systemic administration of losulazine produced marked dose- and time-dependent decreases in dopamine and norepinephrine concentrations in regions outside the blood-brain barrier (i.e., the median eminence, intermediate lobe, and neural lobe), that were accompanied by an increase in plasma concentrations of prolactin and alpha-melanocyte-stimulating hormone. By comparison, losulazine caused a relatively modest, transient depletion of dopamine and norepinephrine (but not 5 hydroxytryptamine) in regions of the brain protected by the blood-brain barrier (i.e., the striatum, nucleus accumbens, and dorsomedial nucleus of the hypothalamus). In contrast to losulazine, acute systemic administration of reserpine caused a prolonged depletion of dopamine, norepinephrine, and 5 hydroxytryptamine in all brain and pituitary regions examined. These results suggest that regional differences in the response of aminergic neurons to acute administration of losulazine and reserpine reflect differences in the ability of these drugs to penetrate the blood-brain barrier. Chronic systemic administration of losulazine produced a similar decrease in dopamine and norepinephrine in the median eminence, intermediate lobe, and neural lobe, suggesting that tolerance does not develop to the ability of losulazine to deplete catecholamines in these regions. Chronic losulazine administration also decreased dopamine concentrations in the striatum, and norepinephrine concentrations in the dorsomedial nucleus, suggesting that losulazine may have cumulative effects on central catecholamine neurons terminating in these brain regions. PMID- 8236259 TI - Cytochrome P4501A1 mediates the metabolism of 2,3,7,8-tetrachlorodibenzofuran in the rat and human. AB - Previous studies have established that TCDF is rapidly metabolized and excreted in rats and that pretreatment of rats with TCDD increases the rate of hepatic metabolism of this compound. The extrahepatic metabolism of TCDF was investigated to assess which enzyme was involved in the metabolism of this compound. Very little metabolism of TCDF was detected in control microsomes (0.3-3.0 pmol/mg/hr), while TCDF metabolism was increased 40- to 200-fold in TCDD-induced rat liver, kidney, and lung microsomes. Since TCDD induces cytochrome P4501A1 and P4501A2 (CYP1A1 and CYP1A2) in the rat liver but only CYP1A1 in kidney and lung, these results suggest that CYP1A1 metabolizes TCDF. To test this hypothesis, TCDF metabolism was investigated in the presence and absence of selective chemical inhibitors and antibodies to CYP1A1 and 1A2. 1-Ethynylpyrene, a suicide inhibitor of CYP1A1 and antibody to rat CYP1A1, produced a dose-dependent inhibition of TCDF metabolism in TCDD-induced rat liver microsomes. Conversely, 2 ethynylnaphthalene, a suicide inhibitor of CYP1A2 and antibody to rat CYP1A2, had no inhibitory effect on the hepatic microsomal metabolism of TCDF. Together, the results strongly indicate that rat CYP1A1 is the primary enzyme responsible for the metabolism of TCDF. 4-Hydroxy-2,3,7,8-TCDF was also identified as the major TCDF metabolite formed by rat CYP1A1. TCDF was also metabolized by human liver microsomes and recombinant yeast microsomes expressing human CYP1A1 and reductase but not by yeast microsomes expressing human CYP1A2 with or without reductase. A similar HPLC profile of TCDF metabolites was observed with microsomes from human liver and yeast expressing human CYP1A1. However, based on ethoxyresorufin-O deethylase activity, a marker of CYP1A1, the relative rate of TCDF metabolism is about 100-fold greater in TCDD-induced rat liver microsomes than in yeast microsomes expressing human CYP1A1 and reductase. Thus, although TCDF is metabolized by rat and human CYP1A1, the results indicate that there are marked quantitative differences in metabolism which suggest that TCDF will be more persistent in humans. PMID- 8236260 TI - Metabolic activation of butylated hydroxytoluene by mouse bronchiolar Clara cells. AB - Metabolism of BHT (2,6-di-tert-butyl-4-methylphenol) is requisite for its pneumotoxic activities. Previous evidence using microsomal preparations from livers and lungs of mice indicated that cytochrome P450-catalyzed hydroxylation of a tertbutyl group to form 6-tert-butyl-2-(hydroxy-tert-butyl)-4-methylphenol (BHTOH) is the first step in the bioactivation of this compound. Subsequent oxidation of BHTOH produces the quinone methide 6-tert-butyl-2-(hydroxy-tert butyl)-4-methylene-2,5-cyclohexadienone (BHTOH-QM), and this highly reactive electrophile may be directly responsible for the pulmonary effects of BHT. The present study assessed the ability of intact bronchiolar Clara cells isolated from mice, a major site of pulmonary xenobiotic metabolism, to convert BHT to BHTOH-QM. The data demonstrate that BHTOH is, in fact, the principal oxidation product in these cells, and that a substantial portion of this metabolite is oxidized further to the quinone methide. BHTOH was found to be considerably more toxic to Clara cells than BHT, and both toxicity and metabolism were simultaneously depressed by the cytochrome P450 inhibitor SKF 525-A. These results strongly support the hypothesis that BHTOH-QM is the active metabolite that generates acute pneumotoxicity and modulates lung tumor formation. PMID- 8236261 TI - Disposition of inhaled isobutene in F344/N rats. AB - Isobutene (2-methylpropene) (CAS No. 115-11-7) is a gas widely used in the chemical manufacturing industry. As an aid to planning long-term toxicity studies, research was conducted to determine the effect of exposure concentrations on the absorption and metabolism of isobutene in F344/N rats. Male F344/N rats (11-15 weeks of age) were exposed for 2 hr to 0, 40, 400, or 4000 ppm isobutene, and a time-course evaluation of blood levels of isobutene was performed using headspace analysis methods. Blood levels of isobutene were linearly related to exposure concentrations between 40 and 400 ppm but increased in a supralinear fashion at the highest concentration, suggesting that the capacity of the rats to metabolize isobutene had been exceeded. Total uptake, excretion patterns, and metabolic conversions were studied in rats exposed for up to 6 hr to 0, 2, 40, 400, or 4000 ppm [14C]isobutene. Absorption of the inhaled isobutene was approximately 8% up to 40 ppm isobutene, but decreased at the higher concentrations. The amount of isobutene metabolized per ppm.hr of exposure was also linear up to 40 ppm but decreased at higher concentrations. Over 90% of the absorbed isobutene was metabolized at exposure concentrations up to 400 ppm, but the exposure to approximately 4000 ppm isobutene resulted in approximately 20% of the absorbed dose exhaled as the unmetabolized isobutene. Two urinary metabolites were identified as isobutenediol and 2-hydroxyisobutyric acid. Two other urinary metabolites were tentatively identified as sulfate conjugates of isobutenediol. Based on these studies, linear dose-response relationships would be expected in chronic toxicity studies for exposures up to 40 ppm isobutene. Additional studies would be required to determine if repeated exposures would induce higher metabolic capacities in the exposed rats. PMID- 8236262 TI - Binding of the cationic dye, Janus green B, as a measure of the specific surface area of crystalline silica in aqueous suspension. AB - Twelve preparations of crystalline silica, with a wide range of particle sizes, were assayed by a new method, which measures surface adsorption of the cationic dye Janus green B to crystalline silica samples in a buffered aqueous suspension. The same samples were also assayed for total surface area by the Brunauer-Emmett Teller (BET) method of surface adsorption of nitrogen gas. A strong linear correlation was found between the two methods of measurement (r = 0.977). Reproducible specific surface area measurements by the Janus green B adsorption method were made on 2-mg samples using ordinary visible wave-length spectrophotometric equipment, whereas the BET method necessitated sample sizes in excess of 100 mg and more specialized instrumentation. Five size-fractionated preparations from the same Min-U-Sil alpha-quartz sample showed an increase in BET surface area and Janus green B binding per unit weight with decreasing particle size. Among four standard alpha-quartz samples tested, Min-U-Sil 5 and F600 had the lowest specific surface areas, whereas DQ-12 and Chinese standard alpha-quartz had much higher surface areas. The synthetic silica preparations cristobalite and tridymite had intermediate surface areas. Binding by the cationic dye Janus green B is consistent with a surface charge mechanism and provides a useful new technique for the assessment of surface characteristics of crystalline silica samples. Its linear relationship to surface area suggests that the ratio of aqueous surface charge to surface area is constant for different crystalline silica preparations. Comparison of surface areas for different preparations of crystalline silica is important in understanding the relative activities of these preparations in studies on mechanisms of silicosis and silica induced lung cancer. PMID- 8236263 TI - Digestive tract absorption of polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls in a nursing infant. AB - The digestive tract absorption of environmental contaminants is an important but poorly understood parameter in contaminant is an important but poorly understood parameter in contaminant risk assessments. The net absorption of polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls in a nursing infant was measured under natural conditions over 12 days. The levels of the substances in the mother's milk were typical for Germany. It was found that for almost all congeners over 90% of the ingested compound was absorbed. This indicates that the common assumption of 100% absorption in nursing infants is reasonable. No firm conclusions could be drawn regarding the absorption of Cl7- and Cl8DD/F due to high blank levels in the cotton diapers used. PMID- 8236264 TI - Variation in antioxidant enzyme activities in anatomic subcompartments within rat and rhesus monkey lung. AB - Antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase, and glutathione S-transferases are thought to be the primary cellular defense against reactive oxygen species. Since pulmonary injury produced by oxidant air pollutants like ozone is highly focal, involving primarily the trachea and centriacinar areas of the lung, measurements of alterations in antioxidant enzyme activities in whole lung may substantially underestimate changes occurring in target areas of the respiratory tract. We have applied a technique for preparation of lung specimens from well-defined anatomic locations to determine whether the focal injury associated with ozone exposure is related to an uneven distribution of antioxidant enzyme activity in the respiratory tract. Our study compared enzyme activities in rat and monkey, species which differ considerably in sensitivity to ozone-induced injury (monkey > rat). The activities of glutathione S-transferase varied less than twofold between different airway subcompartments for both the rat and monkey. Pulmonary veins had approximately 50% of the activity of airways in both species. Glutathione peroxidase activity was slightly higher in proximal compared to distal airways of the rat but was evenly distributed at all airway levels in the monkey. In both species, activity in pulmonary veins was lower than that in airways. The activity of superoxide dismutase was similar in rat and monkey and marked differences were not observed in the various subcompartments studied. Similarly, catalase activity was relatively evenly distributed in rat airways but, in the monkey, the distal bronchiole and lobar bronchus had marginally higher activity than the trachea. We conclude that: (1) measurement of antioxidant enzyme activities in anatomic subcompartments within the lung is feasible using microdissected specimens, (2) antioxidant enzyme activity can vary in different subcompartments of the lung of the same species, (3) the pattern of variation in enzyme activity differs by the enzyme and by species, and (4) species and subcompartment differences in ozone injury are not due primarily to differences in the distribution of antioxidant enzyme activity. PMID- 8236265 TI - Time-dependent effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on serum and urine levels of malondialdehyde, formaldehyde, acetaldehyde, and acetone in rats. AB - The ability of TCDD to produce lipid mobilization and alter lipid metabolism is well known. Previous studies have indicated that TCDD induces an oxidative stress and enhances lipid peroxidation. However, the products resulting from altered lipid metabolism in response to TCDD have not been studied. We have examined the time-dependent changes in serum and urine levels of malondialdehyde, formaldehyde, acetaldehyde, and acetone in response to a single oral 50 micrograms/kg dose of TCDD in rats. The changes in these four metabolic products were quantitated by high-pressure liquid chromatography (HPLC). The effects of TCDD were compared in ad libitum-fed control animals and pair-fed animals. Serum and urine levels of the four metabolites were assayed on Days 0, 3, 6, 9, and 12. Following TCDD administration, significant increases in the four metabolites present in serum and urine were observed at all time points. For example, on Day 6 post-treatment malondialdehyde, formaldehyde, acetaldehyde, and acetone increased approximately 1.8-, 1.4-, 2.0-, and 3.0-fold in serum, respectively, and 1.3-, 1.4-, 1.6-, and 2.0-fold in urine, respectively, relative to pair-fed control animals. Increases in the serum and urine levels of the four metabolites were significantly greater for TCDD animals than for pair-fed control animals at most time points. When the serum levels of malondialdehyde as determined by HPLC were compared with the results obtained by the thiobarbituric acid (TBA) colorimetric method, similar time courses were observed although higher results were obtained for the less specific TBA method. The results clearly demonstrate that TCDD causes markedly elevated serum and urine levels of four specific products associated with lipid metabolism. PMID- 8236266 TI - Nephrotoxicity in rats following liver transplantation from cadmium-exposed rats. AB - Although kidney is considered as the critical organ for cadmium (Cd) toxicity, little is known about the transport of Cd to kidney after chronic exposure. In order to study this transfer, male Lewis rats (150-200 g) were given eight injections (sc) of CdCl2 (3 mg Cd/kg) over 2 weeks which resulted in increases of tissue Cd and metallothionein (MT) concentrations (223 and 1850 micrograms/g, respectively, in the liver and 118 and 873 micrograms/g, respectively, in the kidney). Livers from Cd-injected rats were transplanted to age-matched control healthy Lewis rats and the recipient rats were killed at 2 to 47 days after transplantation. The levels of Cd and MT in the liver of recipient rats were decreased (106 and 1503 micrograms/g, respectively) with time after surgery. On the other hand, renal Cd and MT levels were markedly increased (195 and 1468 micrograms/g, respectively) and most of the Cd in the kidney was bound to MT. About 100 ng/ml of Cd and MT were detected in the plasma of recipient rats by ELISA. There was some periportal fibrosis in the liver due to transplant procedure which did not anastomose hepatic arteries. There was an increase in blood urea nitrogen levels in rats transplanted with Cd-containing liver. In addition, both necrosis and inflammation were observed in the epithelial cells in the proximal tubules in the kidney which typically occurs in chronic Cd toxicity. These results suggest that the major source of renal Cd in chronic Cd exposure may be derived from hepatic Cd which is transported in the form of Cd-MT in blood plasma. PMID- 8236267 TI - Reversibility of lead-induced depression of growth. AB - The overall objective of this investigation was to determine whether the growth depressive effects of lead (Pb) are reversible. The animal model used was the female weanling rat. In Study 1, one group of animals was exposed to Pb for 10 days, a second group was exposed for an additional 21 days, and a third group of animals served as controls. Animals whose Pb was terminated after 10 days caught up completely with controls in terms of linear growth but only partially in terms of ponderal growth. There was no growth catch-up vis-a-vis controls among animals whose Pb exposure was continued for an additional 21 days, to the end of the study. In Study 2, Pb was terminated after only 3 days of exposure. Depression of body weight gain was completely reversed within 1 day after termination of exposure. In Study 3, the role of undernutrition alone (food restriction) was investigated. Depression of body weight gain was immediately apparent and, as with Pb, decrease in linear growth (tail length) was not apparent until later. Catch-up to controls in terms of weight was achieved within 1 day of free access to food when food restriction was for 3 days. Catch-up was also complete after 10 days of food restriction, but was slower (7 days). Evidence for catch-up in terms of linear growth was equivocal. It is concluded that the reversibility of the effects of Pb on growth is dependent on the duration of Pb-induced reduction of food consumption and that linear growth depression is more readily reversible than ponderal growth depression. A final study (Study 4) was undertaken to assess the role of reduced water consumption in limiting food intake and growth. Comparison groups were (1) control, (2) Pb via drinking water, and (3) Pb via sc osmotic pumps. Water consumption was significantly decreased only with Pb by drinking water. Thus, the reduced water consumption with Pb delivered orally seems not to be causally related to the associated depression of food consumption and growth. PMID- 8236268 TI - Comparative effects of two polychlorinated biphenyl congeners on calcium homeostasis in rat cerebellar granule cells. AB - Some polychlorinated biphenyls (PCBs) have been reported to alter locomotor activity and decrease brain dopamine function in laboratory animals. PCBs with ortho- and/or parachlorine substitutions and varying number of chlorinations are known to decrease cell dopamine content in vitro and have been detected in brains of animals exposed to PCBs, suggesting that the neurotoxicity could be mediated by ortho-substituted congeners. Dopamine or other neurotransmitter uptake and release phenomena are dependent on the maintenance of intracellular Ca2+ homeostasis, and perturbations in Ca2+ homeostasis could lead to altered cell function and/or death. We compared the effects of two PCB congeners on Ca2+ homeostasis in cerebellar granule cells: 2,2'-dichlorobiphenyl (DCBP), a putative neurotoxic congener, and 3,3',4,4',5-pentachlorobiphenyl (PCBP), a presumed nonneurotoxic congener. In cerebellar granule cells (6-8 days in vitro), DCBP was cytotoxic as indicated by a significant increase in LDH leakage at 200 microM after 2 hr of exposure and at 100 microM after 4 hr exposure. PCBP, on the other hand, did not affect LDH leakage even at 200 microM for up to 4 hr. Although both congeners increased cerebellar granule cell [Ca2+]i, DCPB was more effective in increasing [Ca2+]i to a greater extent than PCBP. The increase in [Ca2+]i produced by both congeners was not transient, but a steady rise was observed with time. To understand cellular Ca(2+)-buffering capacity, Ca2+ sequestration and Ca2+ extrusion were studied in mitochondria, microsomes, and synaptosomes, isolated from adult rat cerebellum. DCBP was a potent inhibitor of 45Ca2+ uptake by mitochondria (IC50 = 6.17 +/- 0.53 microM) and microsomes (IC50 = 7.61 +/- 0.35 microM). PCBP inhibited Ca2+ sequestration by mitochondria (68% of control) and microsomes (72% of control), but the effects were much less than those produced by equivalent concentrations of DCBP. Synaptosomal Ca(2+)-ATPase was inhibited by DCBP, but not by PCBP. These results indicate that at concentrations where cytotoxicity in cerebellar granule cells was not observed, DCBP increased intracellular [Ca2+]i, and at the same concentrations, Ca2+ sequestration by intracellular organelles and Ca(2+)-ATPase in synaptic plasma membrane were inhibited. Although PCBP increased [Ca2+]i in cerebellar granule cells to some extent, it was not potent in affecting Ca2+ sequestration or Ca2+ extrusion in adult cerebellar components. Hence, PCBP-induced slight increase of [Ca2+]i levels in the cells might have been associated with effective Ca2+ sequestration by intracellular organelles, as seen in cerebellar preparations. The results of this study support the hypothesis that the position of chlorine substitution on the biphenyl ring and/or number of chlorine substitutions may have significant implications for predicting potential effects of PCB congeners in the nervous system, and perturbations in Ca2+ homeostasis might play a significant role in the neuroactivity of PCBs. PMID- 8236269 TI - Application of Cellular Systems in Drug Metabolism and Toxicity Studies. Proceedings of a meeting. Saarland, Germany, July 21-24, 1991. PMID- 8236270 TI - V79 Chinese hamster cells genetically engineered for cytochrome P450 and their use in mutagenicity and metabolism studies. AB - V79 Chinese hamster cells are being genetically engineered for stable expression of cytochromes P450 using a SV40 early promoter containing plasmid as the eukaryotic expression vector. V79 cells lack endogenous cytochrome P450 activity. Therefore, genetically engineered V79 cell lines are defined for the cytochrome P450 isoform acquired upon cDNA mediated gene transfer. V79 cells have a longstanding tradition as indicator cells for various biological endpoints, e.g. mutation, chromosomal aberration, cytotoxicity. The genetically engineered V79 cells combine these useful biological endpoints with metabolic competence. In this sense, V79 cell lines genetically engineered for stable expression of cytochromes P450 present newly developed tools for studying and understanding metabolism related problems in toxicology and pharmacology. PMID- 8236271 TI - Drug and xenobiotic glucuronidation catalysed by cloned human liver UDP Glucuronosyltransferases stably expressed in tissue culture cell lines. AB - Two human UDP-Glucuronosyltransferase (UGT) cDNA clones were stably integrated into V79 chinese hamster fibroblast cells and the functional enzymes were expressed in this heterologous environment. More than 100 drugs and xenobiotics were used as substrates for glucuronidation, catalysed by the cloned UGTs to determine the chemical structures accepted as substrates. UGT HP1 exhibited a limited specificity for planar phenolic compounds, whereas UGT HP4 was more promiscuous in acceptance of non-planar phenols, anthraquinones, flavones, aliphatic alcohols, aromatic carboxylic acids, steroids and many drugs of varied structure. These conclusions are illustrated here by using a series of alkyl- and halophenols. This work indicates the considerable potential value in use of these recombinant cell lines to study human drug glucuronidation. PMID- 8236272 TI - A comparative study of the use of primary Chinese hamster liver cultures and genetically engineered immortal V79 Chinese hamster cell lines expressing rat liver CYP1A1, 1A2 and 2B1 cDNAs in micronucleus assays. AB - Liver microsome preparations (S9 mix) have been extensively used for in vitro genotoxicity studies to provide the capacity for the activation of indirect genotoxins. However, the use of S9 preparations with mammalian cell cultures has raised considerable toxicity problems which limit their use to exposure times which are only a small fraction of the cell cycle. In addition, false negative results may be obtained if reactive metabolites are unable to penetrate the cell membrane or have short half-lives. The generation and detection of a promutagen within a single cell would therefore be advantageous. To this end, we have studied the bioactivation of a panel of promutagens (benzo[a]pyrene, cyclophosphamide, 2-aminoanthracene and sterigmatocystin) in low passage Chinese hamster fibroblasts of hepatic origin (LiC2 cells) and in a series of V79 Chinese hamster cell lines genetically engineered to express rat liver cytochrome P450 cDNAs. These include strains XEM2 (expresses CYP1A1), SD1 (CYP2B1) and strains XEMd-MZ and XEMd-NH which express CYP1A2. The end point selected for study was the induction of micronuclei. The protocol incorporated a cytochalasin B-induced cytokinesis block and the enumeration of micronuclei in the resulting binucleate cells which have undergone one nuclear division following the induction of chromosome damage. Micronuclei containing whole chromosomes and chromosome fragments were distinguished by the use of CREST antibody specific for kinetochore protein as a measure for the presence of centromeres. Micronuclei were induced by the test agents in low passage liver fibroblasts and in immortal V79 cultures only in the presence of Aroclor-induced S9 preparations. The data obtained from micronucleus assays of the genetically engineered V79 cell lines demonstrated the utility of each strain for the optimal detection and quantification of the activity of the individual test compounds. Kinetochore antibody demonstrated differences in the kinetics of induction of micronuclei containing chromosome fragments and whole chromosomes with chemicals such as benzo[a]pyrene. As part of this cytogenetic study, we also conducted karyotypic analyses and spindle fidelity assays of the V79 cell lines to investigate the presence of chromosomal instabilities which may arise as a consequence of the genetic engineering procedure. Such studies represent an important quality control step in the validation of the suitability of each cell line prior to their use in genotoxicity studies. PMID- 8236273 TI - Biotransformation of methylxanthines in mammalian cell lines genetically engineered for expression of single cytochrome P450 isoforms. Allocation of metabolic pathways to isoforms and inhibitory effects of quinolones. AB - V79 Chinese hamster cells genetically engineered for stable expression of single forms of rat cytochromes P450IA1, P450IA2, P450IIB1, human P450IA2, and rat liver epithelial cells expressing murine P450IA2 were used to allocate metabolic pathways of methylxanthines to specific isoforms and to test the suitability of such cell lines for investigations on drug interactions occurring at the cytochrome expressed. The cell lines were exposed to caffeine and/or theophylline and concentrations of metabolites formed in the medium were determined by HPLC. Caffeine was metabolized by human, rat and murine P450IA2, resulting in the formation of four primary demethylated and hydroxylated metabolites. However, there were differences in the relative amounts of the metabolites. The human and the mouse P450IA2 isoforms predominantly mediated 3-demethylation of caffeine. The rat cytochrome P450IA2 mediated both 3-demethylation and 1-demethylation of caffeine to a similar extent. The results support the hypothesis that caffeine plasma clearance is a specific in vivo probe for determining human P450IA2 activity. Addition of the quinolone antibiotic agents pipemidic acid or pefloxacin, both known to inhibit caffeine metabolism in vivo and in human liver microsomes, reduced formation rates of all metabolites of caffeine in cells expressing rat and human P450IA2. Theophylline was mainly metabolized via 8 hydroxylation. All cell lines tested were able to carry out this reaction, with highest activities in cell lines expressing rat or human P450IA2, or rat P450IA1. PMID- 8236274 TI - Rat hepatocyte cultures and co-cultures in biotransformation studies of xenobiotics. AB - Long-term cultures of hepatocytes could represent a suitable in vitro model for biotransformation studies of xenobiotics. At present however, no ideal culture system can be proposed since, in all existing models, phenotypic changes occur, affecting selectively some components of phase I and/or phase II xenobiotic metabolism. From the authors' own results and recent studies of several other investigators, carried out on rat hepatocytes, it becomes clear that four groups of factors may affect biotransformation capacity: soluble medium factors, extracellular matrix components, cell-cell interactions and factors affecting replication. For the maintenance of liver-specific functions, it seems of utmost importance that the tridimensional shape of the hepatocytes is kept. Usually, phase II enzymatic activity is better kept than that of phase I. The cytochrome P450 dependent monoxygenases, in particular, are easily lost. Interesting is the observation that co-cultures of rat hepatocytes with rat liver epithelial cells exhibit higher and much better preserved phase I and phase II biotransformation than monocultures. Clearly, further research is needed to improve this promising in vitro model. PMID- 8236275 TI - Use of human hepatocyte cultures for drug metabolism studies. AB - Among the in vitro models developed to investigate drug metabolism isolated hepatocytes have become the most powerful model. Human hepatocytes can be prepared from whole livers and surgical wedge biopsies. When placed in culture they retain their specific drug metabolizing activities including inducible cytochrome P450 enzymes for several days. Primary human hepatocyte cultures are now increasingly used for studying drug behavior during preclinical development, e.g. drug interactions, stereoselective drug metabolism and drug metabolic profiles. As a rule there is a good in vivo/in vitro correlation in drug biotransformation activity. The main metabolites found in vivo are recovered in vitro. However, quantitative differences are frequently observed. PMID- 8236276 TI - Expression of human cytochrome P450 enzymes in yeast and bacteria and relevance to studies on catalytic specificity. AB - Heterologous expression systems can be utilized to great advantage in the study of cytochrome P450 (P450) and other enzymes involved in the biotransformation of drugs and other xenobiotics. The list of studies made possible with the technology includes discernment of catalytic specificity, elucidation of structure-activity relationships, and various biophysical measurements. There are advantages and disadvantages to each of the vector systems and choices must be made on the basis of needs. Yeast expression systems were used to establish that different P450 2C enzymes are involved in the hydroxylations of tolbutamide and (S)-mephenytion. P450 3A4 was also expressed in yeast and its very broad catalytic specificity was confirmed. Recently, it has been possible to express P450 3A4 as well as other human and animal P450s in bacteria after slight modification of their 5'-coding sequences. PMID- 8236277 TI - Myeloperoxidase-mediated activation of xenobiotics by human leukocytes. AB - Peripheral blood leukocytes contain a variety of enzymes that are capable of metabolising xenobiotics. The enzyme myeloperoxidase (MPO) appears to be the most important for drug metabolism. MPO is a peroxidase/oxidase and generates the powerful oxidant hypochlorous acid. MPO- or MPO-generated oxidants are capable of oxidizing a wide variety of compounds and a broad range of functional groups, especially those that contain nitrogen and sulfur. Leukocytes have a role in immune response; therefore, reactive intermediates generated by leukocyte metabolism of xenobiotics may have a role in idiosyncratic drug reactions, particularly those that are immune-mediated such as drug-induced lupus or agranulocytosis. PMID- 8236278 TI - Prostaglandin-H synthase containing cell lines as tools for studying metabolism and toxicity of xenobiotics. AB - Prostaglandin-H synthase (PHS) can oxidize many xenobiotics and carcinogens (chemicals) in vitro and has been suggested to serve as an alternative metabolic activation enzyme, particularly in tissues low in monooxygenase activity. This article briefly describes types of PHS-catalyzed xenobiotic oxidations and discusses its determinants in cells. Methods employed for studying the involvement of PHS in the bioactivation of chemicals are reviewed with special emphasis on a cell culture system derived from ram seminal vesicles which has been used in studies on the metabolism and the genotoxicity of diethylstilbestrol. PMID- 8236279 TI - Heterologous expression of drug-metabolizing enzymes in cellular and whole animal models. AB - In this report we describe the heterologous expression of glutathione S transferase (GST) and cytochrome P450 reductase (Red) in E. coli and Salmonella typhimurium. The same expression vectors could be applied to both systems and high levels of catalytically active GST and Red were obtained. Interestingly the level of expression was invariably higher in S. typhimurium. The level of the alpha class GST being up to 20% of the total bacterial protein. A further advantage of the salmonella system is that strains were used which can be applied to mutagenicity tests. This system was validated by demonstrating increasing mutation frequency of halogenated hydrocarbons in strains expressing the GST and increased cytotoxicity of mitomycin C in cells expressing P450 reductase. PMID- 8236280 TI - Recombinant yeast in drug metabolism. AB - The usefulness of cDNA-directed expression of human hepatic P450s in yeast for the in vitro study of drug metabolism is emphasized. The major advantages of yeast expression are: (i) relatively high yields of heterologous P450 (approximately 5-10 nmol/l of culture medium) can be obtained; (ii) the expressed P450s are directly active in yeast microsomes, allowing the determination of specific catalytic activities of individual isoforms, which is a prerequisite for the prediction of metabolic pathways for new drug candidates; (iii) transformed yeast microsomes can also be used to study the specific affinity of individual P450s for various substrates and the formation of P450-metabolite complexes by difference visible spectroscopy; such studies can help to predict drug interactions. The advantages of expression in yeast with respect to biochemical studies of drug metabolism are illustrated with data about P450 NF25 (P450 3A4), the major form of human liver. Expressed P450 NF25 is obtained in a functionally active state, and some specific catalytic activities observed in liver microsomes could be reproduced directly with transformed yeast microsomes. The use of genomically modified yeast strains coexpressing human cytochrome b5 and/or overexpressing yeast P450-reductase allowed us to optimize these catalytic activities. In particular, this coexpression system was useful in the study of the in vitro formation of a P450 NF25 Fe(II)-RNO complex. Such inhibitory complexes have been implied in numerous drug interactions involving P450 3A4. PMID- 8236281 TI - Immunotoxicology and expression of human cytochrome P450 in microorganisms. AB - Drug-induced hepatitis can be caused by an abnormal immunological response. In the case of tienilic acid- and dihydralazine-induced hepatitis, we postulated a scheme in which a P450 produced a reactive metabolite (step 1); this reactive metabolite bound to the P450 producing it (step 2) leading to a neoantigen triggering the immune response (step 3); the autoantibodies produced during the immune response recognized the P450 producing the reactive metabolite (step 4). The use of microorganisms (yeast or bacteria) expressing cloned human P450 helped in proving some steps of this postulated scheme, particularly steps 1 and 4. PMID- 8236282 TI - Saccharomyces cerevisiae: an alternative source for human microsomal liver enzymes and its use in drug interaction studies. AB - Heterologous expression of human cDNAs in the yeast Saccharomyces cerevisiae represents an attractive alternative source of human enzymes and allows metabolic studies to be performed without the need of human tissue. Here we report on the functional expression of human microsomal epoxide hydrolase (hmEH) and cytochrome P450 1A1 and 1A2 in yeast. Microsomal fractions of corresponding yeast strains exhibited enzyme specific activities which allowed the characterization of the heterologous enzymes. The use of these microsomes enabled us to study drug interactions on the respective enzymes with pharmacologically relevant drugs such as carbamazepine epoxide, valpromide and ketoconazole. PMID- 8236283 TI - Molecular biology of human xenobiotic-metabolizing cytochromes P450: role of vaccinia virus cDNA expression in evaluating catalytic function. AB - Mammalian xenobiotic-metabolizing cytochromes P450s are membrane-bound enzymes that use O2 and electrons from NADPH to oxidize their substrates. For most chemical substrates, stable metabolites are produced that are destined for further metabolism and elimination from the cell. These enzymes are also capable of metabolically-converting promutagens and procarcinogens to their active proximate metabolites that can kill and transform cells. The xenobiotic metabolizing P450s reside with three distinct families of the large P450 super family. To study the catalytic activities of P450s, particularly human P450s that cannot be easily purified, a cDNA expression system was developed using vaccinia virus. P450 cDNAs incorporated into this lytic virus are efficiently expressed into catalytically-active enzymes that can be used to determine substrate specificities of specific human P450s forms. Activation of the hepatocarcinogen aflatoxin B1 was determined using a series of vaccinia virus-expressed P450s establishing that it is metabolically-activated to a DnA-binding derivative by several human P450 forms, albeit to differing extents. PMID- 8236284 TI - Human cell lines, derived from AHH-1 TK+/- human lymphoblasts, genetically engineered for expression of cytochromes P450. AB - We are developing a panel of human B lymphoblastoid cells which have been engineered to express specific human cDNAs for cytochrome P450 and other xenobiotic metabolizing enzymes. The recipient cells are of a human B lymphoblastoid cell line, designated AHH-1 TK+/-. These cells are transfected using two extrachromosomal vectors both containing OriP sequences derived from Epstein Barr virus but containing independent means of selection in mammalian cells. Using this system, the level of cDNA expression is nearly always stable and consistent from one transfection to another. Thus, once the level of expression has been characterized, cell lines with potentially interesting combinations of xenobiotic-metabolizing enzymes can be predictably developed. cDNAs encoding the following human enzymes have been expressed in this system: CYP1A1, CYP1A2, CYP2A6, CYP2B8, CYP2C6, CYP2C9, CYP2D6, CYP2E1, CYP3A4 and microsomal epoxide hydrolase. We have expressed all of these enzymes individually and have developed cell lines which express combinations of the xenobiotic metabolizing enzymes. The expression of multiple enzymes is important for generalized use of engineered cells as toxicology screening tools. We have primarily used the cell lines in applications to toxicology focusing on procarcinogen activation as detected in assays for the induction of gene locus mutations. In this chapter we discuss the general properties of the system and applications to toxicology testing. PMID- 8236285 TI - [The characteristics of treating the early stages of caries in pregnant women by using laser phoresis]. PMID- 8236286 TI - [The structural characteristics of the masticatory apparatus and dental prophylaxis]. AB - Maxillofacial anthropometric parameters (Pont's index, Tons' index, length of the anterior section of upper dental arch, ratio of upper dental arch to sizes of 12 teeth, ratio of apical basis width to sizes of 12 teeth, mandibular angle size, facial width index) were examined in 59 residents at the town of Omsk born there aged 20 to 25 with normal orthognathic occlusion and low intensity of carious involvement. The data helped define specific features of dental caries prevention measures in this population. PMID- 8236287 TI - [The use of tricalcium phosphate in the vital subtotal extirpation of the pulp]. AB - The authors have proved the desirability of vital subtotal extirpation of the pulp in experiments and clinically. Exposure to tricalcium phosphate transformed the terminal portion of the root pulp into osseocement biologic filling, obturated the apical opening and thus prevented the inflammation in the periodontium and protected it from pathologic exposures. PMID- 8236288 TI - [The morphology of the microorganisms from the contents of a periodontal pocket in relation to the severity of the periodontitis]. AB - Ninety-five patients with periodontitis of slight and medium severity were examined, 21 of them without concomitant diseases, 44 with essential hypertension, and 30 with diabetes mellitus. Besides clinical signs of periodontal abnormalities, the examinations revealed using phase-contrast microscopy four groups of microorganisms in the periodontal pouch contents: cocci, immovable and movable bacilli, and tortuous forms. Multidimensional analysis of clinico-laboratory data revealed a relationship between the counts of some microorganisms and severity of periodontal pathology. PMID- 8236289 TI - [The intensity of protein biosynthesis in the periodontium]. AB - Protein biosynthesis intensity in clinically healthy gingival tissues is the highest at the age 20-30 and reduces by 40-50. In generalized periodontitis this parameter glows, particularly so in exacerbations. An exacerbation is characterized by similar rises of protein synthesis activity in various degrees of the disease. In chronic periodontitis biosynthesis intensity is the minimal in slight and maximal in severe generalized periodontitis. Increased content of sulfhydryl groups in the gingiva in generalized periodontitis does not depend on the disease severity in an exacerbation; in a chronic course their level is the maximal in slight and minimal in severe generalized periodontitis. PMID- 8236290 TI - [Functional hyperemia of the periodontium during the use of chewing gum]. AB - Functional hyperemia developing in the periodontium after 30 sec chewing of a half of a standard chewing gum plate was studied by a noninvasive quantitative method. In subjects with clinically normal periodontium and normal tonic tension of the regional vessels the peak blood flow develops within the first minute of chewing and further vasomotions restore in 30 min. In changed vascular tone, particularly so in periodontal diseases, hyperemia develops persisting for at least 30 min. PMID- 8236291 TI - [Gingival hyperplasia induced by nifedipine]. PMID- 8236292 TI - [Amben and galascorbin in the combined treatment of ulcerative-necrotic lesions of the oral mucosa]. AB - Clinical efficacy of ambene-galascorbic solution was studied in 58 patients with ulcerative necrotic lesions of the buccal mucosa. The results were assessed by clinical and laboratory parameters. Local application of ambene-galascorbic complex was found to be safe and effective in patients with ulcerative necrotic lesions of the buccal mucosa of various origin. PMID- 8236293 TI - [The surgical treatment of periradicular cysts of the jaws]. AB - The author has succeeded in improving functional results of surgical treatment of the teeth that caused pararadical cysts. This was due to access to the pathologic focus, preservation of the 'causative' tooth root, filling of the residual osseous cavity with formalin-treated allogenic placental transplant. A total of 79 patients were operated on using this method. Late results were followed up for 2 years, no recurrences occurred and functional status of the operated on 'causative' teeth permitted their use as abutment teeth for bridge dentures. PMID- 8236294 TI - [The prevention and treatment of patients with suppurative-inflammatory diseases of the maxillofacial area by using vibromassage at autoresonance frequencies]. AB - Autoresonance frequency vibromassage sessions were administered to patients with pyoinflammatory processes in maxillofacial soft tissues. Comprehensive examinations of these patients were carried out to assess the efficacy of this treatment modality; the examinations included rheographic and ultrasonic examinations of blood flow intensity, analysis of neutrophilic activity by chemiluminescence amplitude, measurements of trypsin inhibitor levels and cholinesterase activity, assessment of wound exudate pH values and of resorption capacity of purulent wound tissues. Microbiologic, histologic, cytologic, clinical and biochemical studies were carried out. The findings permit a conclusion that autoresonance vibromassage enhanced biologic purification of the wound from pyonecrotic mass and accelerated reparative regeneration. PMID- 8236295 TI - [The dynamic calcitonin and parathyroid hormone levels of the blood in patients with acute and chronic odontogenic osteomyelitis of the mandible]. PMID- 8236296 TI - [A treatment method for chronic parenchymatous parotitis]. AB - A method for therapy of chronic parenchymatous parotitis is suggested supplementing dimethyl sulfoxide. As reported, the parotid glands produce parotin, an insulin-like substance, whose production is reduced in chronic parotitis; hence, short-acting insulin administered in microdoses was chosen for therapy. To potentiate local insulin effect and increase the sensitivity of oral mucosa peripheral receptors to it a 5% calcium pantothenate solution was used. This method was used in the treatment of 42 patients with chronic parenchymatous parotitis aged 23 to 62. The method is effective, it can be easily used by the patients themselves, and there are virtually no contraindications against such therapy. The authors have applied for inventors' certificate, the priority certificate is No. 4836436/14 as of June 27, 1990. PMID- 8236297 TI - [The practical importance of assessing the immune status of patients with complicated and uncomplicated mandibular fractures]. AB - The authors sum up the practically important general regularities in changes of a number of immunologic parameters associated with the clinical picture of the processes coursing after mandibular fractures. The knowledge of these regularities will help the physicians reduce the incidence of complications by improving the immunologic reactivity of the body using a variety of immunity modulators. PMID- 8236298 TI - [The results of research on the morphological asymmetry of the human maxilla and mandible]. PMID- 8236299 TI - [A clinico-genetic study of patients with congenital deformities of the jaw bones]. AB - Clinical and genetic examination of 78 patients, 43 of these women with jaw bone deformations aged 16 to 30, has revealed that inferior macrognathia combined with superior macrognathia were the most incident in jaw bone deformations. The share of familial cases was 23.9%, this proving a hereditary predisposition to such conditions. These data indicate an etiologic and pathogenetic heterogeneity of patients with deformations of the jaw bones. PMID- 8236300 TI - [Periodontal function and nonspecific resistance in the dynamics of experimental hypo- and hyperthyroidism]. AB - Morphologic study of white rat periodontal tissue and assessment of nonspecific resistance of the body by whole blood antibacterial activity, skin bactericidal activity, blood serum and mixed salivary lysozyme activity were carried out over the course of experimental hypo- and hyperthyrosis starting from day 5 to day 90. The detected pathologic changes in the periodontium were in line with the time course of nonspecific resistance of the body. In hypothyrosis an earlier depression of nonspecific resistance factors was parallelled by earlier and more marked inflammatory destructive processes in the rat periodontium as compared to that in hyperthyrosis. PMID- 8236301 TI - [Photoplethysmographic studies of the vessels of the skin during the elimination of maxillofacial defects with the use of expanders]. PMID- 8236302 TI - [The morphology of the facial skull in patients with active acromegaly based on lateral-view teleroentgenograms]. AB - Measurements of skull base, maxilla and mandible were carried out from lateral teleroentgenograms of the skull made in 27 patients with active acromegaly. Total anterior facial height and mandibular branch height were found the most increased both in men and women. Bone changes of the skull base and facial skeleton in patients with active acromegaly did not depend on the disease duration. PMID- 8236303 TI - [A method for preparing a resin-bonded fixed denture with parapulpar pins]. AB - The authors suggest a method for making an adhesive permanent denture with parapulpar dowels consisting in the use of a whole cast conductor for drilling parapulpar channels in the patient's oral cavity. The conductor is cast simultaneously with the prosthesis of the same metal, hence metal shrinkage in casting is the same, this being a guarantee of the denture accuracy. PMID- 8236304 TI - [The functional restoration of the organs of the oral cavity in complete loss of the teeth]. AB - Examinations of 24 subjects with complete loss of teeth have shown that the functions of various oral organs recover after fitting with complete removable dentures in various periods, from 3 to 60 days. A control group consisted of 18 subjects with intact dentition. The authors suggest assessing adaptation to complete removable dentures from a complex of parameters, including taste sensitivity, salivary gland functional activity, masticatory test, and mandibular movement in this test. PMID- 8236305 TI - [Orthodontic treatment in the distal displacement of the mandible]. AB - A total of 598 patients with various occlusions and dentition defects were examined; 112 (18.7%) of these presented with distal dislocation of the mandible. Shortening of interalveolar distance was found to play an important role in the pathogenesis of this dislocation. This shortening occurs in abnormal teeth abrasion and loss of many molars and premolars maintaining the occlusion height. Masticatory function was found disordered in 56 (50% of patients with distal dislocation of the mandible. Orthodontic treatment of these patients was carried out by sagittal shift of the mandible to a distance of 1 to 2 mm and subsequent fitting with prostheses. Good results were attained in 94.55% of cases. PMID- 8236306 TI - [The cephalometric characteristics of the facial skeleton and base of the skull in prognathic bite]. AB - Lateral teleroentgenography was used to examine facial skeleton and skull base in 62 patients with progenic occlusion formed because of adenoids. Thirty-four subjects with similar occlusions but without nasopharyngeal diseases were examined for control. Specific features of the cerebral and facial segments of the skull associated with the presence of adenoids were detected, namely, reduced size of the anterior and posterior sections of skull base, increased inclination angle, reduced length of maxillary body base and of apical basis and the distal position of the maxilla, increase of the maxillary body anterior height and of the central segment of the face, excessive growth of mandibular body and particularly of its branch parallelled by its increased angle and anterior height of the face, disordered correlations between various parameters. The authors come to a conclusion that progenic occlusion formation in adenoids results from early synostosis of synchondroses at the skull base and from disharmonious development of the entire facial skeleton. PMID- 8236307 TI - [Caries prevalence in preschool children in industrial cities]. AB - Caries incidence was analyzed in preschool children attending kindergartens living in two industrial towns differing by the level and specific features of air pollution and by fluorine levels in potable water. Caries incidence was found to be higher in a ferrous metallurgy center with very low (0.1 to 0.3 ml/l) fluorine levels in potable water. Passive smoking factor was found to contribute to development of caries in children, particularly so in girls. Motor car transport effects on caries development in children were not confirmed. Recommendations on reduction of caries development intensity are suggested. PMID- 8236308 TI - [The pathogenetic validation of the use of the helium-neon laser in treating acute herpetic stomatitis in children]. PMID- 8236309 TI - [The status of the teeth and periodontal tissues of children living in an area polluted by the wastes from an aluminum plant]. AB - The author compares dental, periodontal, and bone system status of children living in two regions characterized by different degrees and patterns of environmental pollution. Children living in regions polluted by aluminum plant waste (sulfur dioxide, nitrogen dioxide, carbon monoxide, dust, hydrogen fluoride) developed specific abnormalities because of body poisoning with fluorides, namely, dental fluorosis, osteoporosis and osteosclerosis of the bones; periodontal diseases in these children were more incident than in controls. These findings prompted the development of a complex of sanitary, technologic, health, therapeutic and prophylactic measures aimed at reduction of fluorine levels in the environment to the normal level and of dental diseases incidence among children living near the Tajik aluminum plant. PMID- 8236310 TI - [Experience in the dispensary observation of children with birth trauma to the maxillofacial area]. PMID- 8236311 TI - [Wound infection in the surgical treatment of children with congenital cleft palate]. AB - Clinico-microbiologic examinations of 153 children with cleft palate aged 2 to 9 were carried out. Two groups of patients with different pattern of operative wound healing after uranoplasty were singled out: 110 ones with primary healing and 13 ones with infectious inflammations of the wound. The incidence of S. aureus carriership on the palatal mucosa before surgery was higher among patients who developed postoperative inflammations. On day 3 after the operation gram negative bacilli were isolated from the majority of patients with postoperative wound inflammations coursing in the presence of marked dysbacteriosis as against patients in whom the wounds healed by primary intention. The authors suggest a method for wound infection prevention in uranoplasty, consisting in irrigation of the operative wound with acilact (a biopreparation) suspension and shortening of antibiotic prevention course to just 48-72 h; the efficacy of this method was confirmed by a reduction of the incidence of postoperative complications. Normal microflora representatives predominated in the microbiocenosis of the operative wound on days 3 and 10-12 after uranoplasty. PMID- 8236312 TI - [The treatment of diastema by using a device with a shape-memory effect]. AB - An orthodontic device for diastema elimination is suggested; the translocated teeth are forced by a TiNi device fit with a shape memory. Due to shape recovery effect occurring in contact warming to 36.6 degrees of the device with preset characteristics it 'strives' to assume the preset shape. The treatment is carried out in accordance with the plan, the shape memory device operates according to a program designed on sham jaws. The results of the treatment were good. PMID- 8236313 TI - [The comparative characteristics of the temporomandibular joints in the rabbit and man]. AB - Examination of rabbit temporomandibular joints has yielded new data contradictory to current notions accepted in dentistry. There are articular tubercles in the temporomandibular joints of rabbits, belonging to rodents, and these joints are incongruent, like in a man. Incongruence of rabbit joints permits combined mandibular movements. The presence of articular tubercles and occlusion curves in the rabbit is in line with an assumption of the compensation theory on their relationship and interdependence. These data permit a conclusion that rabbit temporomandibular joints are in many respects similar to those of man. PMID- 8236314 TI - [The efficacy of using an Iruxol ointment in treating alveolitis]. AB - Analysis of the results of treatment of 150 alveolitis patients has demonstrated the advantages of the method using iruxol ointment (75 patients) as against the routine one (reference group--75 patients). Therapy with this ointment resulted in complete arrest of the painful syndrome in 2.11 +/- 0.15 days, whereas in the reference group it was attained in but 5.40 +/- 0.18 days. The temperature reaction normalized in all the test group patients within 3 days, whereas in the reference group such results were attained in only 74.36% of the patients. Inflammatory symptoms in the paramaxillary soft tissues were liquidated with iruxol ointment in 5.96 +/- 0.14 days, whereas in routine therapy this was observed in only 7.46 +/- 0.18 days. PMID- 8236315 TI - [The use of the electric bulb from UAZ-469 automobiles in the lamps of US-30-01 dental units manufactured by the Media (Sofia) medical apparatus plant]. PMID- 8236316 TI - [Hypertrophic gingivitis of drug origin (a review of the literature)]. PMID- 8236317 TI - [The psychological characteristics of middle-aged and elderly patients and their consideration in the process of preparing dentures (a review of the literature)]. AB - Experience gained in orthodontic treatment of elderly patients and analysis of the papers of Russian and foreign authors helped us demonstrate the usefulness of knowledge of an aging man psychology to make up and maintain contacts with him or her. Recommendations are given concerning policy with elderly patients at different stages of treatment and a relationship singled out between personality traits of an orthodontist and his (her) ability to psychologically influence the patients. PMID- 8236318 TI - [The conducting of practical games during postgraduate instruction in departments of physician advanced training]. PMID- 8236319 TI - [On the centenary of the formation of the Society of Dentists and Dental Surgeons in Moscow]. PMID- 8236320 TI - [The prevention and treatment of local radiation reactions and complications (injuries) in the maxillofacial area (a lecture)]. PMID- 8236321 TI - [V. M. Uvarov and his contribution to the development of the scientific problems in maxillofacial surgery and dentistry (on the centenary of his birth)]. PMID- 8236322 TI - [Impressions of a visit to the Center for Plastic Surgery in Miami (USA)]. PMID- 8236323 TI - Synthesis and 1H and 13C nuclear magnetic resonance of 16-methylene-17 alpha hydroxypregna-1,4,9(11)-triene-3,20-dione. AB - A three-step synthesis of 16-methylene-17 alpha-hydroxypregna-1,4,9(11)- triene 3,20-dione, which has been previously obtained by microbial transformation, was achieved from 3 beta-hydroxy-16 alpha, 17-epoxy-16 beta-methyl-5 alpha-pregn 9(11)-en-20-one in an overall yield of 35%. All compounds involved in the process were characterized spectroscopically and their 13C NMR shielding effects are briefly discussed. PMID- 8236324 TI - H-2 gene complex and corticosteroid responsiveness: evidence that the corticosteroid hormone signal transduction pathway in the adult mouse lung is not associated with haplotype-specific responses to corticosteroids. AB - Differential responsiveness to corticosteroids (CORT) has been shown to be related to HLA haplotype. A strong association between the mouse homolog to the human HLA complex, the H-2 complex, and intrauterine responses to CORT have also been demonstrated; haplotype differences alter CORT-induced susceptibility to cleft palate and temporal differences in lung maturation. Since variation in the glucocorticoid receptor (GR) is associated with tissue specific responses to CORT, we hypothesize that haplotype-specific CORT responsiveness may be regulated by H-2 associated modification of GR expression and/or function. Given that H-2 congenic mice are genetically identical except at the H-2 complex on mouse chromosome 17 and the GR structural gene is encoded on chromosome 18, the GR gene is identical in these mice. However, any step in the GR signal transduction pathway may be regulated by gene(s) at or near the H-2 complex and result in haplotype-specific differences in CORT responsiveness. We have investigated differences in qualitative and quantitative characteristics of the adult B10 (H 2b) and B10.A (H-2a) pulmonary GR by Scatchard analysis, immunochemical and biochemical assays. No differences in the GR binding parameters (BMAX and Kd), receptor form and level, or ligand-GR complex binding to glucocorticoid response element (GR-GRE) were detected, leading us to conclude that H-2 associated factors do not regulate the relative intrauterine responses to CORT by modulating the adult GR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236325 TI - Comparative effect of synthetic aminoestrogens with estradiol on platelet aggregation. AB - The in vitro effect upon platelet aggregation of estradiol and synthetic estrogens (prolame, buame, and proacame) is described. Prolame and buame produced a dose-dependent inhibition on platelet aggregation. Estradiol and proacame did not show anti-aggregating effects. The results suggest that the use of prolame and buame in estrogen therapy could reduce the risk of thrombo-embolic accidents. PMID- 8236326 TI - A new 5 alpha,8 alpha-epidioxy sterol from the Okinawan marine sponge of the Axinyssa genus. AB - A new sterol (axinysterol) was isolated from the Okinawan marine sponge of the genus Axinyssa. The structure of axinysterol was assigned as 5 alpha,8 alpha epidioxyergosta- 6,22,25-trien-3 beta-ol based on spectroscopic analysis and chemical transformation. PMID- 8236327 TI - Synthesis and biochemical studies of 7 alpha-substituted androsta-1,4-diene-3,17 diones as enzyme-activated irreversible inhibitors of aromatase. AB - Several 7 alpha-thiosubstituted derivatives of androstenedione have demonstrated effective inhibition of aromatase, the cytochrome P450 enzyme complex responsible for the biosynthesis of estrogens. Introduction of an additional double bond in the A ring resulted in 7 alpha-(4'-amino)phenylthioandrosta-1,4-diene-3,17-dione (7 alpha-APTADD), a potent inhibitor that inactivated aromatase by an enzyme catalyzed process. Additional 7 alpha-thiosubstituted androsta-1,4-diene-3,17 dione derivatives were designed to further examine enzyme-catalyzed inactivation. Two halogenated and one unsubstituted 7 alpha-phenylthioandrosta-1,4-diene-3,17 diones were synthesized via an acid-catalyzed conjugate Michael addition of substituted thiophenols with androsta-1,4,6-triene-3,17-dione. Two 7 alpha naphthylthioandrosta-1,4-diene-3,17-diones were synthesized via either acid catalyzed or based-catalyzed conjugate Michael addition of substituted thionaphthols with androsta-1,4,6-triene-3,17-dione. These agents were evaluated for aromatase inhibitory activity in the human placental microsomal preparation. Under initial velocity assay conditions of low product formation, the inhibitors demonstrated potent inhibition of aromatase, with apparent Ki's ranging from 12 to 27 nM. Furthermore, these compounds produced time-dependent, first-order inactivation of aromatase in the presence of NADPH, whereas no aromatase inactivation was observed in the absence of NADPH. This enzyme-activated irreversible inhibition, also referred to as mechanism-based inhibition, can be prevented by the substrate androstenedione. Thus, the apparent Ki values for these inhibitors are consistent with earlier studies on 7 alpha-substituted competitive inhibitors that indicate bulky substituents can be accommodated at the 7 alpha-position.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236328 TI - 1,4-addition reaction with thiols and conformational analysis with PM3 molecular orbital calculations of 19-oxygenated androst-4-ene-3,6,17-triones. AB - Reactions of androst-4-ene-3,6,17-trione (1) and its 19-hydroxy or 19-oxo derivative (2 or 3), suicide substrates of aromatase, with thiols were initially studied. Treatment of 4-ene-3,6-diones 1-3 with benzyl-mercaptan in MeOH at room temperature gave the corresponding 4 alpha-benzylthio-5 alpha-androstane-3,6 diones (4-6) as the major products in 24-80% yields. The C18 steroid, estr-5(10) ene-3,6,17-trione (7), was also isolated on the treatment of 19-oxo steroid 3. Oxidation with NaIO4 and reduction with Raney Ni of the adducts gave the corresponding 4-ene-3,6-dione and desulfurized products, respectively. The results show that 19-oxygenated steroids 2 and 3 react with a thiol in a 1,4 addition manner. By means of PM3 molecular orbital calculations, the conformational features of the 19-oxygen functions of 4-ene-3,6-diones 2 and 3, 5 alpha-3,6-diones 10 and 11, and their 4 alpha-methylthio derivatives 14 and 15, model compounds of 1,4-adducts 5 and 6, were determined. In the compounds examined, the 19-hydroxy steroids favor a conformation having the hydroxyl group above the A-ring, whereas the 19-oxo substituent is oriented in the out-of-ring position (not above either A- or B-ring). These calculations suggest that compound 1 would inactivate aromatase by the same steric course of the oxygenation at C-19 as that of the natural substrate, androstenedione. PMID- 8236329 TI - Direct quantitative digital autoradiography of testosterone metabolites in the pilosebaceous unit: an environmentally advantageous trace radioactive technology. AB - Androgen metabolism is one of the major keys for a better understanding of conditions such as androchronogenetic alopecia, acne, and other androgen-related skin disorders. This paper addresses the process by which testosterone metabolism leads to the following major androgens: androstenedione, dihydrotestosterone, 3 alpha- and 3 beta-androstanediol, androsterone, epiandrosterone, and androstanedione. Our report describes a methodology developed for the direct quantitative measurement from the silica gel plate of these metabolites. After detailing the chromatographic procedures to achieve the complete separation of the seven testosterone metabolites on a single plate, specifications are given for obtaining accurate measurements by 1) calibration of the radiodetectors and 2) internal and external standardization of samples and plates. Analytical criteria are discussed in terms of comparison of the level of sensitivity, reproducibility, and practicability obtained by both the linear analyzer and the direct digital autoradiograph. Signals as weak as 25 dpm were easily detected and calibration curves were obtained for the range of 50-500 dpm. For biological measurements the coefficients of variation do not exceed 10%. Given the difficulty of obtaining large amounts of microdissected subfractions of the pilosebaceous unit and the necessity of evaluating the complete pattern of testosterone transformed into its 3 alpha,3 beta-, 5 alpha-reduced and 17 beta dehydrogenated secondary derivatives, digital autoradiography appears to be a powerful yet simple tool for studying androgen metabolism. In addition, this methodology offers an important environmental advantage: the high sensitivity of the detectors makes it possible to minimize the quantities of radioactive materials that must be handled or discarded. PMID- 8236330 TI - Enhanced androgen sensitivity in serum-free medium of a subline of the LNCaP human prostate cancer cell line. AB - The LNCaP-Fast Growing Colony (FGC) human prostate cancer cell line proliferates in response to the addition of dihydrotestosterone (DHT) 10(-10)-10(-8) M in charcoal-stripped serum-supplemented media. LNCaP-FGC cells will not attach or proliferate in serum-free conditions. LNCaP-FGC stock cultures were maintained in medium supplemented with 10% FBS and added DHT (10(-9) M) for > 25 passages (6 months). The resultant subline was designated as LNCaP-ss (supersensitive) because of its ability to attach in serum-free medium and to proliferate in response to very low levels of DHT. LNCaP-ss cells were grown in serum-free medium and proliferation assessed after 2, 3, 5, and 7 days' treatment with DHT. Significant enhancement of growth was demonstrated after 7 days' treatment with DHT over a wide range of concentrations (DHT 10(-15)-10(-7) M) with maximal stimulation (3 x control, p < .001) noted with DHT 10(-14) M. Changing the medium during the course of the experiment decreased, but did not eliminate, the DHT induced cellular proliferation. Scatchard analysis of binding studies with LNCaP ss cells revealed that both the Kd for the androgen receptor (AR) and the number of AR sites/cell were similar to the corresponding values reported for the parental line. AR mRNA levels in LNCaP-ss cells, as measured by RNase protection assay, were significantly down-regulated by 7 days' treatment with DHT 10(-15), 10(-13), and 10(-9) M.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236331 TI - Various formulae of androstanediol glucuronide. PMID- 8236332 TI - Headache in stroke. AB - BACKGROUND AND PURPOSE: The etiology of headache in stroke is not known, and its relation to migraine and tension-type headache is unclear. The aim of our study was to investigate and classify headache appearing in stroke patients prospectively, using the new headache classification as determined by the Headache Classification Committee of the International Headache Society (1988). METHODS: Two hundred eighty consecutively admitted patients aged younger than 81 years with acute stroke were examined and questioned about headache and prior headache complaints; 238 (85%) were able to communicate. RESULTS: Sixty-five (27%) of the 238 patients experienced headache from 3 days before to 3 days after stroke. Headache occurred in 50% of patients with intracerebral hemorrhage, in 26% with infarction, and in 15% of patients with lacunar infarction. Headache was more common when stroke occurred in posterior circulation than anterior circulation (P < .02). Fifty-six patients were able to give further information about headache characteristics. The headache in thromboembolic stroke was classified as tension-type headache (25 patients), migraine-like headache (14 patients), and other headache (12 patients). Migraine was more frequent in vertebrobasilar stroke. Headache was lateralized in 33% of cases. In patients with unilateral headache and unilateral stroke lesion, the headache was ipsilateral in 14 of 17 cases. In infarction, severity of headache showed no relation to lesion size or lesion localization. Patients with previous tension type headache and migraine experienced reactivation of known headache equally often. CONCLUSIONS: (1) Headache occurs in one fourth of patients with acute stroke. (2) Unilateral headache is usually ipsilateral to stroke lesion. (3) Headache severity is not related to size of ischemic stroke lesion. PMID- 8236333 TI - Apathy following cerebrovascular lesions. AB - BACKGROUND AND PURPOSE: Although apathy has been reported to constitute a frequent sequela of stroke lesions, there have been no prospective studies on the frequency and correlates of apathy after stroke lesions. In the present study, we examined the frequency and correlates of apathy in a consecutive series of 80 patients with cerebrovascular lesions. METHODS: We included patients within the first 10 days after a stroke lesion. Patients were examined with a comprehensive neuropsychiatric battery that included the Apathy Scale. RESULTS: Eighteen patients (22.5%) showed apathy, nine of whom were also depressed. On the other hand, 18 patients (22.5%) showed depression in the absence of apathy. Although depression and apathy may exist independent of one another, major depression (but not minor depression) was associated with an increased frequency of apathy. Apathy was also significantly associated with older age, cognitive impairments, and deficits in activities of daily living. Finally, apathy was significantly associated with lesions in the posterior limb of the internal capsule. CONCLUSIONS: These findings demonstrate that apathy is a frequent finding among patients with acute stroke lesions and may coexist with important emotional and cognitive poststroke disturbances. PMID- 8236334 TI - Incidence of silent cerebral infarction in patients with major depression. AB - BACKGROUND AND PURPOSE: There have been few studies of the incidence of silent cerebral infarction detected by magnetic resonance imaging in patients with presenile or senile major depression. METHODS: We examined silent cerebral infarction in patients with presenile and senile major depression who were diagnosed at Hiroshima Prefectural Hospital. The diagnostic criteria of the American Psychiatric Association (DSM-III-R) were used. Patients with stroke or focal neurological symptoms were excluded. RESULTS: Silent cerebral infarction was observed in 51.4% of the patients with presenile-onset presenile depression, and the incidence was significantly higher than in patients with juvenile-onset presenile depression (P < .01). Among the patients with senile major depression, silent cerebral infarction was observed in 65.9% of those with presenile-onset depression and in 93.7% of those with senile-onset depression. CONCLUSIONS: Our findings suggest that half of presenile-onset major depression and the majority of senile-onset major depression might be organic depression related to silent cerebral infarction. Because major depression occurring for the first time during or after the presenile period may be related to silent cerebral infarction, it is important to keep this possibility in mind when treating such patients. PMID- 8236335 TI - Stroke in a Saudi Arabian National Guard community. Analysis of 500 consecutive cases from a population-based hospital. AB - BACKGROUND AND PURPOSE: We sought to determine the crude incidence rate, patterns, and risk factors associated with different types of stroke in a defined Saudi population. METHODS: Records of 500 (342 male, 158 female) consecutive patients with first-ever stroke admitted from December 1982 to June 1992 in a hospital that exclusively serves the Saudi Arabian National Guard community were reviewed. Diagnosis was confirmed by brain computed tomography, and the most likely etiology was determined on the basis of relevant clinical, radiological, and laboratory data. RESULTS: The mean age of the patients was 63 +/- 17 years. Males predominated in all types of stroke (P < .001). The crude annual incidence rate was 43.8 per 100,000. Ischemic strokes accounted for 76.2%, and these included 52% with large and 24.2% with lacunar infarctions. Intracerebral hemorrhage was detected in 21.4%, whereas subarachnoid hemorrhage was rare (2.4%). Hypertension (56%), diabetes mellitus (42%), and cardiopathy (33%) were common risk factors. Sixty-one patients (12%) died during the first month after their stroke. CONCLUSIONS: The study suggests that stroke incidence is low in Saudi Arabia compared with industrialized countries, which could be because of the predominance of young age groups. The overall distribution of stroke types was closer to that of Western populations than to the Japanese, in whom hemorrhagic strokes are highly prevalent. However, the high combined frequencies of lacunar infarctions and intracerebral hemorrhages suggest that disease of the small cerebral arteries played a more important role in Saudis than in Western populations. PMID- 8236336 TI - Trends in survival of hospitalized stroke patients between 1970 and 1985. The Minnesota Heart Survey. AB - BACKGROUND AND PURPOSE: Age-adjusted stroke mortality rates declined approximately 50% between 1970 and 1990 in both the United States and Minnesota, but the reasons for this decline are not clear. This report examines possible improvements in short- and long-term survival of hospitalized definite stroke patients in the Minneapolis-St Paul (the Twin Cities) metropolitan area during this period. METHODS: Fifty percent random samples of patients discharged with an acute stroke diagnosis from area hospitals were selected in 1970 (n = 1200), 1980 (n = 1040), and 1985 (n = 896). Trained nurses abstracted pertinent clinical data from the hospital charts. By standardized clinical criteria similar to World Health Organization criteria (without computed tomography data), 376, 442, and 453 definite strokes were established for 1970, 1980, and 1985, respectively. RESULTS: Age- and sex-adjusted 28-day case fatality of definite stroke improved significantly from 1970 to 1985; the odds ratio (OR) of death within 28 days in 1985 (versus 1970) patients was 0.55 (95% confidence interval [CI], [0.39, 0.77]). Substantial improvements in 28-day mortality were observed both from 1970 to 1980 and from 1980 to 1985, although the latter change was not statistically significant. Further adjustment for predictors of early stroke mortality (such as level of consciousness) somewhat attenuated these results. Age- and sex-adjusted 5-year survival of definite stroke also improved significantly from 1970 to 1985 (OR, 0.72; 95% CI, [0.54, 0.96]), although the improvement was restricted to the 1970 to 1980 time period (OR, 0.76; 95% CI, [0.57, 1.01]). None of the survival trends differed significantly between men and women. CONCLUSIONS: There were marked improvements in survival from 1970 to 1985 among hospitalized stroke patients in the Twin Cities. These improvements occurred almost exclusively in the acute hospitalization phase. Although the advent of computed tomography and improvements in hospital record-keeping during this period prevent an unequivocal conclusion, improved medical care and decreased severity of stroke probably contributed to gains in survival. PMID- 8236337 TI - Impact of early surgery on outcome after aneurysmal subarachnoid hemorrhage. A population-based study. AB - BACKGROUND AND PURPOSE: Population-based patient materials have not been used earlier in assessing the effects of neurosurgical treatment on survival and functional outcome of subarachnoid hemorrhage. Moreover, the proportion of all subarachnoid hemorrhage patients who might be candidates for neurosurgical treatment has not been estimated. METHODS: We compared the survival and functional outcome of two population-based patient materials from Central Finland in 1976 through 1978 (n = 146) and 1980 through 1987 (n = 351). The most important basic characteristics of both materials were similar. In the 1970s, only patients aged < 60 years with carotid territory aneurysms were operated on after an interval of 2 weeks from the bleeding. In the 1980s, early surgery was attempted, and the other exclusion criteria were abandoned. Allocation to medical or surgical treatment was not randomized. RESULTS: During the 1970s, only 14% of the patients had surgical treatment, with a median delay of 15 days after the bleeding; in the 1980s, the corresponding figures were 46% and 4 days. Despite these fundamental changes in the treatment policy, the survival up to 3 years in the 1980s was only marginally improved compared with the 1970s. Conversely, the functional outcome at 4 years after the bleeding was significantly better in the 1980s than the 1970s, with 82% and 64% of the survivors, respectively, being independent in the activities of daily living (P = .002). We estimated that 60% of all patients with subarachnoid hemorrhage might be candidates for neurosurgical treatment, provided that there are no delays in admission or evaluation. CONCLUSIONS: An active treatment policy of subarachnoid hemorrhage including early surgery only marginally improves survival, but the quality of life of the survivors is significantly better. Only 60% of all patients in the population with subarachnoid hemorrhage can, at least theoretically, benefit from surgical treatment. PMID- 8236338 TI - Ischemic and hemorrhagic stroke in patients on oral anticoagulants after reconstruction for chronic lower limb ischemia. AB - BACKGROUND AND PURPOSE: Information on the long-term fate of patients with chronic lower limb ischemia is limited. We investigated the long-term risk of the first ischemic and hemorrhagic cerebral stroke in patients on long-term anticoagulant therapy after reconstruction for chronic limb ischemia. METHODS: In a retrospective study, 376 consecutive patients were seen at regular intervals according to a standard protocol. Only 3 (0.7%) were lost during follow-up (mean duration, 5.9 years). Anticoagulation was with coumarin derivatives followed by prothrombin times periodically. Primary end points were ischemic and hemorrhagic cerebral stroke events, which were confirmed by CT scan, autopsy, or operation in 85% of the cases. Major vascular events were analyzed as a composite secondary end point. The influence of several clinical variables on these outcome events was evaluated in univariate and multivariate analyses. RESULTS: Thirty-nine patients (10%) had 41 stroke events (23 ischemic, 18 hemorrhagic); 22 of these patients (56%) died from stroke. The cumulative ischemic stroke risk was 5% at 5 years and 12% at 15 years. Prior myocardial infarction was the only independent predictor (relative risk [RR], 3.1; P < .05). The cumulative hemorrhagic stroke risk was 3% at 5 years and 17% at 15 years. Systolic hypertension (RR, 4.8; P < .01) and insulin-dependent diabetes mellitus (RR, 5.4; P < .01) were significant and independent predictors. The risk for a major vascular event was 29% at 5 years and increased to 56% at 15 years. Independent predictors were advanced age (RR, 1.4; P < .005), insulin-dependent diabetes (RR, 2.2; P < .005), and prior myocardial infarction (RR, 1.8; P < .01). CONCLUSIONS: Patients with chronic lower limb ischemia, notably those with prior myocardial infarction, are at high risk for ischemic stroke. Those with systolic hypertension or insulin-dependent diabetes mellitus are at high risk for hemorrhagic stroke. PMID- 8236339 TI - Transhemispheric passage of microemboli in patients with unilateral internal carotid artery occlusion. AB - BACKGROUND AND PURPOSE: Ischemic episodes distal to an internal carotid artery occlusion are common. We undertook this study to look for evidence of transhemispheric passage of embolic material in this patient category as a mechanism for embolic events. METHODS: Seven symptomatic patients with unilateral internal carotid artery occlusion and contralateral stenosis were examined by transcranial Doppler ultrasonography with 2-MHz probe (average monitoring time, 2.5 hours per patient). Both middle cerebral arteries and (if present) the reverse-flow anterior cerebral artery ipsilateral to the occluded internal carotid were monitored. Three patients were reexamined 1 month after carotid endarterectomy. RESULTS: Embolic signals were detected in the middle cerebral artery ipsilateral to the stenosed internal carotid artery in all seven patients and in the opposite middle cerebral artery in four patients. In these four patients, a reverse-flow anterior cerebral artery was observed in which embolic signals were detected. No embolic signals were detected after surgery in any of the three patients who underwent carotid endarterectomy. CONCLUSIONS: Transhemispheric passage of embolic material occurs in patients with unilateral internal carotid artery occlusion and contralateral stenosis. Endarterectomy of the stenosed internal carotid artery may eliminate the detected embolic signals in both hemispheres. Transcranial Doppler ultrasonography could be used as a diagnostic tool to identify the embolic source in patients with unilateral carotid occlusion. PMID- 8236340 TI - Computerized detection of cerebral emboli and discrimination from artifact using Doppler ultrasound. AB - BACKGROUND AND PURPOSE: Transcranial Doppler ultrasound can detect circulating cerebral emboli. Monitoring of patients with potential embolic sources may allow identification of high-risk patients who can then be selected for prophylactic treatment. However, practical patient monitoring will require automated programs that can detect emboli and differentiate them from artifact. METHODS: A new off line algorithm for the detection of emboli, which detects the characteristic relative power increase occurring with an embolus, was evaluated in both an animal model and in patients. (1) In a sheep model, solid embolic materials (thrombus, platelet aggregates, and atheroma) were introduced into the proximal carotid artery while the distal carotid artery or a major branch was insonated. The signals resulting from 77 emboli (mean size, 1.77 mm) were studied and compared with the Doppler signals resulting from artifact. (2) In patients, 100 embolic signals occurring in three patients were analyzed and compared with signals associated with artifact in the same patients. RESULTS: (1) In the sheep model, emboli resulted in a short-duration, high-intensity signal, but intensity increase alone did not distinguish between emboli and artifact. In contrast, the algorithm discriminated embolus from artifact with a sensitivity of 98.7% and a specificity of 98.0%. (2) In patient studies, embolic signals were differentiated from artifact with a sensitivity of 97.2% and a specificity of 97.0% by the algorithm. CONCLUSIONS: Using such an algorithm, detection of cerebral emboli and discrimination from artifact are possible with a high sensitivity and specificity. Incorporation of such an algorithm into an on-line system should make prolonged patient monitoring practical. PMID- 8236341 TI - Activation of healthy hemisphere in poststroke recovery. A transcranial Doppler study. AB - BACKGROUND AND PURPOSE: The possible involvement of the healthy hemisphere in functional recovery in stroke hemiplegia is still controversial. This study of patients who had suffered ischemic stroke assessed circulatory changes in the healthy hemisphere by transcranial Doppler ultrasonography during opposition exercise of fingers that had recovered movement. METHODS: Measurements of flow velocity in the middle cerebral arteries were recorded in 12 recovered hemiplegic stroke patients and 12 healthy sex- and age-matched control subjects during a 2 minute sequential thumb-to-finger opposition task, alternatively performed with normal and recovered fingers. The effects of the motor task on heart rate, blood pressure, and end-tidal PCO2 were also evaluated. RESULTS: With respect to the basal values, a comparable increase of flow velocity (%mean +/- SD) in the contralateral middle cerebral artery occurred in patients during movement of normal (9.7 +/- 2.7%) and recovered fingers (10.4 +/- 5.1%) and in controls (11.3 +/- 4.9%). However, in the middle cerebral artery ipsilateral to the hand performing the motor task, the increase of flow velocity was significantly higher (P < .001, two-way analysis of variance) during movement of the recovered hand in patients (10.5 +/- 4.6%) than during the movement of the normal hand in both patients (1.7 +/- 3.7%) and controls (1.8 +/- 4.5%). No significant difference of changes in heart rate, blood pressure, and end-tidal PCO2 was observed for patients or normal control subjects during the various phases of the study. CONCLUSIONS: Our data suggest that activation of the healthy hemisphere contributes toward functional recovery of motor deficit after stroke and confirms the ability of transcranial Doppler ultrasonography to correlate cerebral artery flow dynamics with selective cerebral activation. PMID- 8236342 TI - Internal carotid artery dissection in a community. Rochester, Minnesota, 1987 1992. AB - BACKGROUND AND PURPOSE: Cervical internal carotid artery dissections are diagnosed with an increasing frequency, but reliable epidemiologic data are not available. The aim of this study was to determine the incidence rate of spontaneous cervical internal carotid artery dissection in a defined population. METHODS: Using the medical record linkage system used for epidemiologic studies in Rochester, Minn, all patients diagnosed with spontaneous cervical ICA dissection for 1987 through 1992 were identified. RESULTS: A total of 10 patients with spontaneous cervical internal carotid artery dissection (6 women and 4 men; mean age, 44 years) were identified. For the period 1987 through 1992, the average annual incidence rate for all ages was 2.6 per 100,000 (95% confidence intervals, 0.9 to 4.2). CONCLUSIONS: This study, for the first time, provides incidence rates for spontaneous cervical internal carotid artery dissections. No diagnoses were made before 1987, probably reflecting an increased awareness of the disorder among physicians. PMID- 8236343 TI - Clinical features of recurrent embolization in acute cardioembolic stroke. AB - BACKGROUND AND PURPOSE: Recurrent embolization is a serious problem in acute cardioembolic stroke. However, the clinical features and predisposing factors of recurrent embolization have not been fully elucidated. METHODS: Subjects were 227 consecutive patients (128 men and 99 women, aged 68.6 +/- 13.2 years) with acute cardioembolic stroke who did not receive anticoagulant therapy during the first 14 days after stroke onset. We assigned the subjects to two groups according to the occurrence or nonoccurrence of recurrent attacks within 14 days of the stroke onset. We assessed their clinical features, coagulation study results, and underlying heart disease. RESULTS: Recurrent brain or systemic embolization during the first 14 days after onset was noted in 46 patients (20.3%, group A) but not in the other 181 (group B). Recurrent embolization was more frequently noted at an early phase than at a late phase during the initial 14 days. Mortality was higher in group A (19.6%) than in group B (8.8%). The mean plasma level of antithrombin III (77.8 +/- 19.5%) at admission in group A patients was significantly lower than that in group B patients (87.9 +/- 15.5%). After admission, hematocrit decreased in group B patients but slightly increased in group A patients, in whom diuretics were more commonly used. Rheumatic heart disease and prosthetic valves, in addition to the presence of intracardiac thrombi, were seen more commonly in group A patients, whereas atrial fibrillation without organic heart disease and myocardial infarction were more frequent in group B patients. CONCLUSIONS: Low plasma levels of antithrombin III, dehydration, the use of diuretics, and the presence of rheumatic heart disease, prosthetic valves, and intracardiac thrombi seem to be predisposing factors for recurrent embolization. Immediate anticoagulation may be considered in acute cardioembolic stroke patients if such predisposing factors are demonstrated. PMID- 8236344 TI - The impact of cardiac index on cerebral hemodynamics. AB - BACKGROUND AND PURPOSE: Current noninvasive testing allows accurate assessment of cerebrovascular hemodynamics. The cardiovascular influence on the noninvasive assessment of cerebrovascular studies has not been defined. This study was designed to determine the effect of cardiac index (CI) on cerebral blood flow velocities, ocular pulse amplitude, ophthalmic systolic pressure, and ocular blood flow (OBF) as currently estimated by noninvasive laboratories. METHODS: Based on a retrospective study of 181 patients, we prospectively evaluated 45 patients undergoing right heart catheterization for hemodynamic monitoring to correlate the relation between CI, transcranial Doppler sonography, and ocular pneumoplethysmography. Patients with hemodynamic instability, severe carotid stenoses, massive cerebral infarct, or sepsis were ineligible for the study. Simultaneous recordings of systemic blood pressure, ophthalmic systolic pressure, heart rate, ocular pulse amplitude, middle cerebral artery blood flow velocities, and cardiac output were obtained on all patients. OBF was calculated from the heart rate and ocular pulse amplitude. RESULTS: The relation between OBF and CI is expressed by the equation CI = 2.36 + 0.61 x OBF (r = .47, P = .0010). The middle cerebral artery peak systolic velocities and CI had a correlation of .36 (P = .0181). The equation, derived from the linear relation between OBF and CI, was then validated on a sample of 15 patients. With the apparent linear relation between OBF and CI, we used the derived equation to predict CI from OBF. The OBF determination predicted CI within 30% in all patients and within 20% in 53.3% of the patients. CONCLUSIONS: We demonstrated that OBF and middle cerebral artery systolic velocity decrease with diminishing CI. Our findings suggest that CI may be potentially estimated in selected patients by noninvasive assessment of OBF using ocular pneumoplethysmography. PMID- 8236345 TI - Cerebral hypoperfusion in stroke prognosis and brain recovery. AB - BACKGROUND AND PURPOSE: The value of acute cerebral blood flow measurements in stroke prognosis is controversial. No previous study has determined whether acute perfusion deficits independently add to a validated clinical prognostic score. We aimed to compare the value of acute hypoperfusion deficits with a quantitative clinical score in stroke prognosis and to correlate the changes in perfusion with brain recovery. METHODS: Volumetric analysis of regional hypoperfusion was performed in 38 patients with middle cerebral infarction within 72 hours of onset by use of single photon emission computed tomography and 99mTc hexamethylpropylene amine oxime. Stroke severity was assessed by the Canadian Neurological Score and Barthel Index. Allen's prognostic score was determined acutely in all patients. Clinical outcome was evaluated in 36 of 38 patients, of whom 18 had repeat blood flow studies. RESULTS: Acute hypoperfusion correlated with both the outcome Barthel Index (P < .001, r = -.61) and Canadian Neurological Score (P < .001, r = -.64). Allen's score correlated better with both the outcome Barthel Index (P < .001, r = .80) and Canadian Neurological Score (P < .001, r = .81). Acute hypoperfusion deficits, after allowing for Allen's score, independently predicted neurological but not functional outcome. Despite overall neurological improvement, mean hypoperfusion increased on the repeat blood flow studies (P < .05). CONCLUSIONS: Volumetric analysis of acute regional hypoperfusion within 72 hours of onset predicts stroke outcome after 3 months, but Allen's score is a better prognostic method. Neurological recovery is not associated with chronic infarct reperfusion. PMID- 8236346 TI - Cerebellar infarction. Clinical and neuroimaging analysis in 293 patients. The Tohoku Cerebellar Infarction Study Group. AB - BACKGROUND AND PURPOSE: We performed this multicenter study to explore the full spectrum of the clinical characteristics and neuroimaging findings of cerebellar infarction, including patients with mild to severe illnesses. METHODS: We studied 293 consecutive patients with cerebellar infarction diagnosed by computed tomography and/or magnetic resonance imaging who were admitted to 36 hospitals during 5 years. RESULTS: Cerebellar infarcts constituted 2.3% of the total patients with acute brain infarction. The backgrounds and risk factors were similar to those in patients with infarctions of the cerebral hemispheres. At least 24% were embolic, and the diagnosis of embolism could not be ruled out in 27%. Infarcts involving the superior cerebellar artery (SCA) region (52%) and the posterior inferior cerebellar artery (PICA) region (49%) were far more frequent than those involving the anterior inferior cerebellar artery (AICA) region (20%). Patients with SCA infarcts exhibited obtunded consciousness and ataxia more frequently than those with PICA infarcts (P < .05). Infarcts in the PICA regions were associated with abnormalities of the PICA (64%) or the vertebral arteries (57%), whereas infarcts in the SCA and AICA regions were associated with abnormalities in the SCA or AICA, respectively, in approximately 30% of patients, in the basilar artery in approximately 16%, and in the vertebral artery in more than 60% of patients. Outcomes were poorer with SCA infarcts than with AICA and PICA infarcts. CONCLUSIONS: These data indicate similar frequencies of SCA and PICA infarcts and illustrate the difference in clinical presentation and outcomes between SCA and PICA infarcts. They also indicate that not only in situ thrombosis but also cardiogenic or artery-to-artery embolism and the insufficiency of collateral circulation play important roles in the pathogenesis of cerebellar infarction. PMID- 8236347 TI - Neuroprotection by excitatory amino acid antagonist augments the benefit of thrombolysis in embolic stroke in rats. AB - BACKGROUND AND PURPOSE: The effects of delayed thrombolysis with alteplase and neuroprotection with an excitatory amino acid receptor antagonist and their combination were tested in an embolic stroke model. METHODS: In 61 rats the carotid artery territory was embolized with arterial-like fibrin-rich clots. Hemispheric cerebral blood flow before and after embolization was measured by intra-arterial 133Xe injection method. The animals were assigned to one of the following treatments: (1) vehicle-treated controls (n = 15); (2) dizocilpine 1 mg/kg i.v. 5 minutes after embolization (n = 16); (3) alteplase 20 mg/kg as an intravenous continuous infusion starting 2 hours after embolization (n = 16); and (4) both agents (n = 14). Carotid angiography displayed the site of occlusion of the cerebral arterial tree immediately after and 3 hours after embolization, and the clinical neurological score was assessed after the rats recovered from anesthesia and before the rats were killed. Brains were fixed after 2 days and evaluated neuropathologically; infarct volume affecting cortical and deep brain structures was measured separately. RESULTS: Both alteplase and dizocilpine reduced the total infarct volume (P = .05 and P = .04, respectively, Mann-Whitney tests). Dizocilpine reduced the incidence of cortical infarctions by 48% (P < .001, Fisher's test). Only the combined treatment significantly reduced deep brain infarctions (P = .03, Mann-Whitney test). The combined treatment also improved the clinical score by 83% compared with controls, by 75% compared with the group treated by dizocilpine alone, and by 50% compared with the group treated by alteplase alone. Sixty-seven percent of thrombolytic-treated animals recanalized completely compared with 39% of those given no thrombolytics (P = .05, Fisher's test). The clinical outcome correlated with infarct size (P < .01, Spearman test). CONCLUSIONS: Our results document comparable efficacy of delayed thrombolysis and excitatory amino acid receptor antagonism in this model and suggest that combination of these two therapeutic approaches may yield additional benefit in treatment of thromboembolic stroke, particularly in cases where deep brain (end-artery-supplied) structures are affected. PMID- 8236348 TI - Improved posthypoxic recovery of synaptic transmission in gerbil neocortical slices treated with a calpain inhibitor. AB - BACKGROUND AND PURPOSE: Among the various calcium-induced biologic events occurring in hypoxic neurons, activation of the calcium-activated neutral proteinase (calpain) is a likely mediator of neuronal degeneration. In this study, we assessed the protective effects of a calpain inhibitor (Cbz-Val-Phe-H) against hypoxic damage to the neocortex. METHODS: An in vitro neocortical slice model from gerbils was used to study the delay to hypoxic depolarization during hypoxia and the recovery of synaptic responses after hypoxia. These responses were examined in control slices and slices treated with Cbz-Val-Phe-H. RESULTS: The delay to hypoxic depolarization did not differ between treated and control groups. In contrast, synaptic recovery after a fixed period of hypoxia (15 minutes) was significantly improved in the Cbz-Val-Phe-H-treated slices (P < .01). Concentrations of Cbz-Val-Phe-H of 50 mumol/L or greater were significantly more protective than a concentration of 20 mumol/L (P < .01). CONCLUSIONS: The data indicate that calcium-activated proteolysis plays a critical role in hypoxic damage to the neocortex and that calpain inhibitors may be useful therapeutic agents. PMID- 8236349 TI - Subarachnoid blood causes pial arteriolar constriction in newborn pigs. AB - BACKGROUND AND PURPOSE: The present study was designed to determine in newborn animals the delayed effect of subarachnoid blood on pial arteriolar diameter and eicosanoid concentrations in cortical periarachnoid fluid. METHODS: Forty-eight to 96 hours after subarachnoid blood installation, closed cranial windows were implanted over the cerebral area exposed to blood in anesthetized, artificially ventilated newborn piglets. All pial arterioles greater than 60 microns in diameter were measured, and cortical periarachnoid fluid was collected for the determination of eicosanoids. RESULTS: Subarachnoid blood resulted in a 20% to 30% decrease in the average diameter of pial arterioles exposed to blood for 48 to 96 hours, a decreased number of large pial arterioles (greater than 200 microns), and an increased number of small arterioles (60 to 100 microns). No changes in dilator prostanoids (prostacyclin [as 6-keto-prostaglandin F1 alpha] and prostaglandin E2) were detected. Concentrations of vasoconstrictor prostanoids in cortical cerebrospinal fluid increased. Thromboxane B2 increased to 430 +/- 70 pg/mL, and prostaglandin F2 alpha increased to 1370 +/- 180 pg/mL compared with 250 +/- 20 and 860 +/- 70 pg/mL, respectively, in the control group. The concentration of peptidoleukotrienes increased to 400 to 600 pg/mL 72 to 96 hours after blood installation, while the level in the control group was less than 80 pg/mL. CONCLUSIONS: The altered balance between vasodilator and vasoconstrictor eicosanoids could contribute to cerebral vasoconstriction after subarachnoid blood installation in newborn pigs. PMID- 8236350 TI - Mice expressing a bovine basic fibroblast growth factor transgene in the brain show increased resistance to hypoxemic-ischemic cerebral damage. AB - BACKGROUND AND PURPOSE: Cerebral intraventricular infusion of acidic or basic fibroblast growth factor has been shown to attenuate ischemic damage to hippocampal CA1 neurons in the gerbil. The purpose of the present study was to determine if the basic fibroblast growth factor transgenic mouse has an enhanced ability to resist the effects of severe cerebral hypoxemia-oligemia. METHODS: Mice that were transgenic for bovine basic fibroblast growth factor were exposed to right carotid artery ligation, hyperglycemia, and 20 minutes of 1% carbon monoxide. After 5 days' recovery, brains were examined for histological damage. RESULTS: Counts of CA1 neurons in the right hippocampus showed a significantly higher number of neurons per millimeter CA1 in hypoxic-ischemic transgenic mice compared with nontransgenic controls (transgenic, 260 +/- 33; nontransgenic, 151 +/- 37 neurons per millimeter CA1; P < .05). CONCLUSIONS: The results indicate that basic fibroblast growth factor transgenic mice, as judged by CA1 hippocampal neuronal survival, have an enhanced ability to resist the effects of a complex hypoxic-ischemic cerebral insult. PMID- 8236351 TI - Retention of cerebrovascular dilation after cortical spreading depression in anesthetized rabbits. AB - BACKGROUND AND PURPOSE: We examined responses of rabbit pial arterioles to three different stimuli before and after induction of cortical spreading depression. METHODS: In urethane-anesthetized rabbits equipped with a closed cranial window, we measured pial arteriolar diameter during baseline conditions, topical application of calcitonin gene-related peptide (CGRP), topical application of acetylcholine, and inhalation of 10% CO2 in air (arterial hypercapnia) before cortical spreading depression and 30, 60, and 120 minutes after cortical spreading depression. Cortical spreading depression was induced by localized application of a 5% KCl solution anterior to the arteriole being measured. RESULTS: Average baseline diameter was approximately 90 microns. During cortical spreading depression, arteriolar diameter increased to a peak value that was 50 +/- 4% above baseline (n = 32). Before cortical spreading depression, arteriolar diameter changed 47 +/- 7% (n = 9) during hypercapnia, 17 +/- 3% (n = 4) during 10(-9) mol/L CGRP, 42 +/- 10% (n = 7) during 10(-7) mol/L CGRP, 29 +/- 6% (n = 4) during 10(-6) mol/L acetylcholine, and 61 +/- 13% (n = 6) during 10(-4) mol/L acetylcholine. Arteriolar responsiveness to any of these stimuli was not changed significantly by prior cortical spreading depression. CONCLUSIONS: Dilator capacity of pial arterioles is still intact in urethane-anesthetized rabbits after cortical spreading depression. PMID- 8236352 TI - Interleukin-1 beta mRNA expression in ischemic rat cortex. AB - BACKGROUND AND PURPOSE: Interleukin-1 beta is a proinflammatory cytokine produced by blood-borne and resident brain inflammatory cells. The present study was conducted to determine if interleukin-1 beta mRNA was produced in the brain of rats subjected to permanent focal ischemia. METHODS: Rat interleukin-1 beta cDNA, synthesized from stimulated rat peritoneal macrophage RNA by reverse transcription and polymerase chain reaction and cloned in plasmid Bluescript KS+, was used to evaluate the expression of interleukin-1 beta mRNA in cerebral cortex from spontaneously hypertensive rats and normotensive rats subjected to permanent middle cerebral artery occlusion. Interleukin-1 beta mRNA was quantified by Northern blot analysis and compared with rat macrophage RNA standard. To correct for gel loading, blots were also analyzed with cyclophilin cDNA, which encodes an abundant, conserved protein that was unchanged by the experimental conditions. RESULTS: Interleukin-1 beta mRNA produced in the ischemic zone was significantly increased from 6 hours to 120 hours, with a maximum of 211 +/- 24% of interleukin 1 beta reference standard, ie, 0.2 ng stimulated rat macrophage RNA, mRNA compared with the level in nonischemic cortices (4 +/- 2%) at 12 hours after ischemia (P < .01; n = 6). Interleukin-1 beta mRNA at 12 hours after ischemia was markedly elevated in hypertensive rats over levels found in two normotensive rat strains. Neurological deficits were also apparent only in the hypertensive rats. CONCLUSIONS: Brain interleukin-1 beta mRNA is elevated acutely after permanent focal ischemia and especially in hypertensive rats. These data suggest that this potent proinflammatory and procoagulant cytokine might have a role in brain damage following ischemia. PMID- 8236353 TI - Progressive intracranial occlusive disease associated with deficiency of protein S. Report of two cases. AB - BACKGROUND AND PURPOSE: Deficiency of the free fraction of protein S has been associated with arterial or venous stroke. The pathogenesis of vascular occlusion in patients with protein S deficiency is not known. We present two cases of cerebral infarction and deficiency of protein S in which the subjects had progressive intracranial occlusions. CASE DESCRIPTION: A 16-year-old girl was admitted because of left brain stem infarction and protein S deficiency. Cerebral angiography disclosed stenosis of the right intracranial vertebral artery and occlusion of the left posterior cerebral artery. A second angiogram performed 18 months later disclosed occlusion of the right vertebral intracranial artery. In the second case, a 17-year-old girl was admitted because of left hemispheric cerebral infarction and protein S deficiency. Cerebral angiography showed stenosis of the left anterior cerebral artery, left supraclinoid internal artery, and left middle cerebral artery. A second cerebral angiogram performed 5 months later disclosed occlusion of the left anterior cerebral artery and poor hemispheric perfusion through the left middle cerebral artery. CONCLUSIONS: Based on our cases, we postulate that some patients with prothrombotic states may develop progressive intracranial arterial occlusions, possibly secondary to a permanent thrombogenic stimulus. We suggest routinely searching for prothrombotic states in young patients with intracranial occlusion, especially if the occlusion is progressive and other causes are not obvious. PMID- 8236354 TI - Hemiplegia due to posterior cerebral artery occlusion. AB - BACKGROUND: Hemiplegia is a rare manifestation of posterior cerebral artery occlusion. The acute clinical picture may be difficult to differentiate from occlusion of the middle cerebral artery. A mechanism for the hemiplegia has not been conclusively determined. CASE DESCRIPTION: We describe a patient with hemiplegia secondary to posterior cerebral artery occlusion by an embolized fragment of a prosthetic valve. Computed tomographic scan showed the foreign body just distal to the origin of the posterior cerebral artery with infarction of its vascular territory. These findings were later confirmed at autopsy. There was no radiological or autopsy evidence of involvement of the other cerebral arteries or their territories. CONCLUSIONS: The patient provides further evidence that occlusion of the posterior cerebral artery just distal to its junction with the posterior communicating artery may produce contralateral hemiplegia without oculomotor nerve nucleus involvement. PMID- 8236355 TI - Prediction of outcomes after intracerebral hemorrhage. PMID- 8236356 TI - Do anger and aggression affect carotid atherosclerosis? PMID- 8236357 TI - Neurobiology of depression in stroke patients. PMID- 8236358 TI - Carotid endarterectomy with TCD and EEG monitoring. PMID- 8236359 TI - Preliminary studies of the effects of extracorporeal membrane oxygenator on the disposition of common pediatric drugs. AB - There is an increased use of extracorporeal membrane oxygenation (ECMO) in the last 15 years for critically ill neonates. While receiving ECMO therapy, the critically ill infant needs various medications. We performed an in vitro study to evaluate the potential effect of the membrane oxygenator on drug extraction. Two closed ECMO circuits were set up at rates of 320 ml/min. One circuit was new and the other was used clinically for 5 days. Morphine at 8 ng/ml, gentamicin 10 micrograms/ml, vancomycin 40 micrograms/ml, phenobarbital 20 micrograms/ml, and phenytoin 20 micrograms/ml were injected into the circuit at 1-h intervals. Blood samples were drawn from the circuit at 10, 30, 60, and 240 minutes after injection. In the new circuit, drugs were eliminated as follows: vancomycin 36%, gentamicin 10%, phenobarbital 17%, phenytoin 43%, morphine 36%. In the used system, levels fell to a much smaller extent: vancomycin 11%, phenobarbital 6%, gentamicin 0%, phenytoin 0%, and morphine 16%. In a child receiving 20 micrograms/kg/h infusion of morphine, steady-state concentrations of 68.2 ng/ml fell to 11.6 ng/ml after changing the membrane. Our data indicate that the ECMO is associated with lowering of the concentrations of commonly used medications and that this process may depend partially on how new the membrane is. Before these changes may lead to new dosing guidelines for small children receiving ECMO, more experiments with new and used systems are warranted, as well as with different types of ECMO. PMID- 8236360 TI - Distribution of amitriptyline and nortriptyline in blood: role of alpha-1 glycoprotein. AB - To interpret blood levels of tricyclic antidepressants, we studied the distributions of amitriptyline and nortriptyline in human blood and explored their control by plasma factors. Each compound (300 ng/ml) was added to whole adult blood and to cord blood with decreased alpha-1-glycoprotein (AGP). Drugs (250 ng/ml) were also added to washed erythrocytes (RBCs) resuspended in autologous plasma or saline (hematocrit = 0.4) with or without AGP, albumin, or tris(2-butoxyethyl) phosphate (TBEP), used to displace AGP-bound drugs. Plasma AGP was determined in all adult blood donors (n = 17). With adult blood, plasma amitriptyline was 393 +/- 52 ng/ml, RBC amitriptyline was 184 +/- 33 ng/ml. Plasma and RBC nortriptyline were 199 +/- 28 and 288 +/- 39 ng/ml, respectively. With saline, cellular amitriptyline and nortriptyline were 81 +/- 10 and 88 +/- 6%, respectively. With plasma, cellular amitriptyline and nortriptyline were 25 +/- 8 and 49 +/- 10%, respectively. The corresponding cord blood values were 52 +/- 12 and 62 +/- 6%. Graded increments of AGP in saline reproduced the distribution pattern seen with increasing concentrations of plasma. Albumin did not influence drug distribution. TBEP markedly increased erythrocyte amitriptyline in adult but not in cord blood. Plasma AGP correlated positively (p = 0.031) with the RBC/plasma ratio of amitriptyline. Amitriptyline is predominantly distributed in plasma, nortriptyline in RBCs. This differential distribution is dose dependent and reflects the higher binding of amitriptyline to AGP when compared with nortriptyline. Interpretation of tricyclic antidepressant blood levels is clarified by obtaining assays from RBCs and plasma. PMID- 8236361 TI - Effects of plasma lipid levels on blood distribution and pharmacokinetics of cyclosporin A. AB - The present study attempted to characterize the distribution of cyclosporin A (CsA) among the lipoprotein fractions, very-low-, intermediate-, low-, and high density (VLDL, IDL, LDL, and HDL, respectively) in the plasma of patients awaiting heart transplantation and the influence of plasma lipid constituents on the pharmacokinetics of CsA. Major fractions of a therapeutic concentration of CsA were found in HDL and in LDL. In addition, plasma lipid concentrations (total cholesterol, triglycerides, phospholipids, VLDL-cholesterol--TC, TG, PL, VLDLc, respectively) are positively correlated with the CsA distribution within the LDL fraction, and negatively correlated with the CsA distribution within the HDL fraction. Thus, the percentage of CsA in each type of lipoproteins was shown to vary with the lipid levels among individuals. A significant negative correlation was found between apparent distribution volume at steady state (Vss) in plasma and TC, PL, and LDLc and between the area under the curve measured in blood (AUCB) for whole blood and PL. PMID- 8236362 TI - Generation of pharmacokinetic data during routine therapeutic drug monitoring: Bayesian approach vs. pharmacokinetic studies. AB - In three groups (each n = 12) of unselected hospitalized patients treated either with digoxin, theophylline, or gentamicin routinely performed TDM measurement of trough steady-state plasma levels (+ peak levels in case of gentamicin) was combined with a pharmacokinetic study at steady state (multiple blood sampling during one dosing interval). Pharmacokinetic parameters (apparent volume of distribution Vd, total plasma clearance CL) needed for individualization of dosage were evaluated by the Bayesian approach and a model-(in)dependent pharmacokinetic program (TOPFIT). Comparison of both methods revealed some small differences in the pharmacokinetic parameters for all three drugs. Mean deviations of the Bayesian estimates from the pharmacokinetic calculations of the three drugs ranged between 20 and 38% for Vd and between 13 and 22% for CL, indicating that the Bayesian approach provided reliable pharmacokinetic estimates for individualizing drug dosage under routine conditions. Therefore, it is suggested that routine TDM combined with Bayesian-based analyses can be regarded as an alternative to pharmacokinetic studies in clinically relevant populations. PMID- 8236363 TI - Audit of theophylline plasma level monitoring in a pediatric hospital. AB - Theophylline assays performed in a children's hospital were audited, focusing particularly on distribution of plasma concentration values, interpretation of the assay result, appropriateness of the advice conveyed to the physician, and the response to this advice. Assays performed by a multidisciplinary therapeutic drug monitoring (TDM) service during a 1-month period in 1988 and again in 1992 were chosen for review. Theophylline concentration in plasma was quantified by fluorescence polarization immunoassay. Medical technologists interpreted results and made specific recommendations concerning dose changes and/or need for follow up assays on 48.5 (1988 survey) and 37.9% (1992 survey) of results. Review of these recommendations revealed that only approximately 1% could have been improved or were incorrect. Physician compliance with dose change recommendations was similar in both surveys (chi 2 = 0.16, p = 0.7), with recommendations being followed on 75.9 (1988) and 73.3% (1992) of occasions. Compliance with recommendations for ordering additional samples at specific times also was similar between the two surveys (chi 2 = 2.06, p = 0.15), with recommendations being followed on 75 (1988) and 87.2% (1992) of occasions. Overall, combining the data for both surveys, (a) significantly more out-patients than in-patients (69.2 versus 27.8%, respectively) were subtherapeutic, (b) significantly fewer out patients than in-patients (27.6 versus 70.2%, respectively) were within the therapeutic range (chi 2 = 33.7, p < 0.001), but (c) numbers (2-4%) of potentially toxic results within the two groups were similar. PMID- 8236364 TI - In vitro evaluation of a continuous-sampling device for pharmacokinetic parameter estimation. AB - Using an in vitro pharmacokinetic model, area under the curve (AUC) estimates from an osmotic continuous-withdrawal device were compared to AUC estimates by a conventional trapezoidal method. Ten experiments were done under two different conditions: (a) half-life of 1 h (n = 5) and (b) half-life of 2 h (n = 5). Sampling was done for 6 and 12 h, respectively. The AUC estimates from the two methods were highly correlated (r = 0.948, p < 0.0001). The mean coefficient of variation was 9.5% (n = 10), and mean AUC values were not statistically different by analysis of variance (p = 0.4). The sampling device sampled at 0.121 +/- 0.011 ml/h; the mean volume was 0.79 +/- 0.03 and 1.34 +/- 0.02 ml over 6 and 12 h, respectively. Continuous sampling provided a reasonable estimation of AUC, required only one analytical sample, and sampled at a consistent zero-order rate. PMID- 8236365 TI - High-performance liquid chromatography determination of dextromethorphan and dextrorphan for oxidation phenotyping by fluorescence and ultraviolet detection. AB - To establish the usefulness of fluorescence detection to quantify urinary concentrations of dextromethorphan and dextrorphan for oxidation phenotyping, we determined the molar concentration ratio of dextromethorphan to dextrorphan in 38 subjects by UV and fluorescence detection. Dextromethorphan and dextrorphan concentrations were quantified after overnight hydrolysis of urine samples and organic solvent extraction with heptane and butanol. The compounds were separated by high-performance liquid chromatography using a phenyl column and a mobile phase consisting of acetonitrile and an aqueous mixture of 0.01 M heptane sulfonic acid and 0.01 M phosphate buffer. The eluents were detected in series by a UV detector (280 nm) and fluorescence detector (excitation 280 nm and emission 310 nm). The dextromethorphan to dextrorphan molar concentration ratio by UV and fluorescence detection was highly correlated (r = 0.997) and not statistically different (p = 0.1036). However, increased sensitivity with fluorescence detection enabled detection of lower dextromethorphan and dextrorphan concentrations when compared with UV detection. Fluorescence detection was able to detect dextromethorphan as low as 0.02 microgram/ml, which may be helpful in phenotyping individuals with extremely rapid metabolism of dextromethorphan. Fluorescence detection also produced chromatograms with significantly less interference and allows a more accurate quantitation of dextromethorphan and dextrorphan concentrations. PMID- 8236366 TI - Concurrent liquid chromatographic measurement of fluoxetine, amitriptyline, imipramine, and their active metabolites norfluoxetine, nortriptyline, and desipramine in plasma. AB - An expedient and specific liquid chromatographic method for a concurrent measurement of fluoxetine (FLU), norfluoxetine (NFLU), amitriptyline (AMI), nortriptyline (NTRIP), imipramine (IMI), and desipramine (DES) is described. Using a mixture of acetonitrile:methanol:0.056 M ammonium acetate:1 M ammonium hydroxide (100:10:4.5:2.6, by volume) as mobile phase, the compounds along with doxepin (DOX) (internal standard) were separated on a 10 mu, 8 mm x 10 cm C18 Resolve cartridge in conjunction with radial compression liquid chromatographic module, and were detected in the effluent spectrophotometrically at 220 nm. A hexane:isoamyl alcohol (98:2, by volume) solution was used for extraction of plasma and the drugs were removed from the organic phase with 0.03% phosphoric acid prior to injection. Under these conditions, no interference in the assay was observed, and the retention times of NFLU, DOX, FLU, AMI, IMI, NTRIP, and DES were 7.8, 11.6, 16, 17.8, 20.9, 31, and 35 min, respectively. The assay was highly linear (r > 0.994), and the within- and between-day coefficient of variance was consistently < or = 9.8%. This assay is currently being used to simultaneously measure these drugs in patients and to investigate their steady state pharmacokinetics when used in combination. PMID- 8236367 TI - Determination of phenobarbital, ethosuximide, and primidone in human serum by micellar electrokinetic capillary chromatography with direct sample injection. AB - The determination of antiepileptic drugs in human serum by micellar electrokinetic capillary chromatography (MECC) with direct sample injection is discussed. Nanoliter quantities of patient sera are applied to the beginning of a fused silica capillary filled with a phosphate/borate buffer (pH 9.2) containing 75 mM sodium dodecylsulfate. Upon application of an electric field along the capillary, endogenous and drug substances are transported toward the cathode and separate into distinct zones which are detected by on-column UV absorption. Phenobarbital, ethosuximide, and primidone are shown to elute in front of the solubilized proteins, thus permitting quantitation of these drugs without any sample pretreatment. For phenobarbital and ethosuximide, MECC data obtained using the external standard method and peak areas as the basis for quantitation are shown to be in excellent agreement with those of nonisotopic immunoassays and, for ethosuximide, also with those of high-performance liquid chromatography. The correlation coefficients (n = 50) are between 0.972 and 0.986. Intraday and interday reproducibility data are 2.0-4.5% and 4.5-8.0%, respectively. For primidone, insufficient samples have been available for a comprehensive comparison of MECC data with those of other analytic techniques. PMID- 8236368 TI - Determination of total and free carbamazepine and the principal metabolites in serum by high-performance liquid chromatography with photodiode-array detection. AB - A precise and accurate high-performance liquid chromatography (HPLC) method has been established for the simultaneous analysis of carbamazepine (CBZ), carbamazepine-10,11-epoxide (CBZ-E), and trans-10,11-dihydroxy-10,11-dihydro-CBZ (CBZ-H) in serum samples and their ultrafiltrates. CBZ and its metabolites are eluted in a 3-microM ODS-Hypersil column (250 x 2 mm) at a column temperature of 40 degrees C. The mobile phase is a mixture containing potassium phosphate buffer acetonitrile-methanol (110:50:30, vol/vol/vol) at a flow rate of 0.2 ml/min. Signals are monitored by a photodiodearray detector with a main sample wavelength of 215 nm and a bandwidth of 10 nm. Coefficients of variation (CVs) for within- and between-day are within 5%, with the recovery rates ranging from 98.16 to 104.64%. This method has the necessary sensitivity and linearity for routine therapeutic monitoring of both total and free CBZ and its principal metabolites. Total serum concentrations of CBZ, CBZ-E, and CBZ-H obtained from 55 epileptic children were 12.58 +/- 4.42, 2.45 +/- 1.22, and 5.83 +/- 3.17 (mean +/- SD, micrograms/ml), respectively. Levels of free CBZ, CBZ-E, and CBZ-H were 2.59 +/- 0.93, 1.05 +/- 0.57 and 3.73 +/- 1.87, respectively. Free fractions of CBZ, CBZ E, and CBZ-H were 20.98 +/- 4.34, 42.63 +/- 8.21, and 65.41 +/- 7.80%, respectively. CBZ-H and CBZ-E had larger CVs than did CBZ (54.34 and 49.75 vs 35.15%, respectively, for total levels, and 50.31 and 54.46 vs 36.22%, respectively, for free levels), as well as higher free fractions. Determination of both total and free concentrations and free fractions of CBZ and its metabolites, as well as their ratios, should provide additional needed information for therapeutic drug monitoring of CBZ. PMID- 8236369 TI - A sensitive and simple high-performance liquid chromatographic method for the determination of mitoxantrone in plasma. AB - A sensitive high-performance liquid chromatographic (HPLC) method for measuring the anthracene derivative, mitoxantrone, in plasma is described. After protein precipitation with 5-sulfosalicylic acid, samples are resolved by isocratic elution from a C18 reverse phase support and quantified by ultraviolet (UV) detection. Recovery of mitoxantrone after deproteinization was > 70%. Within-run and between-day coefficients of variation (CVs) were 5.1 and 13.7%, respectively, at mitoxantrone concentrations of 20 nM (n = 6). The limit of detection was 2.5 nM and the standard curve linear up to 1 microM. Stability studies have shown that mitoxantrone is stable in spiked whole blood for 3-6 h, provided the samples are kept on ice. The drug is stable in plasma and deproteinized plasma samples for at least 24 h. The method requires few manipulations and is readily adaptable to automation. PMID- 8236370 TI - Reversed-phase high-performance liquid chromatography determination of quinine in plasma, whole blood, urine, and samples dried on filter paper. AB - The analysis of quinine in whole blood, plasma, urine, and samples dried on filter paper is described. Extraction was made with toluene followed by back extraction into phosphate buffer. A reversed-phase liquid chromatography system with fluorescence detection was used. The within-day coefficient of variation of the method was 4-10% at the lower limit of determination (2 nM in plasma and 50 nM in whole blood, dried samples, and urine) and 2-4% at 10 microM. The quinine concentration was found to be lower in whole blood than in plasma (mean ratio, plasma-whole blood, 1.17). The concentration in capillary blood was lower than that in venous blood (mean ratio, capillary blood-venous blood, 0.93). PMID- 8236371 TI - Determination of midazolam and its alpha-hydroxy metabolite in human plasma and urine by high-performance liquid chromatography. AB - We describe a new high-performance liquid chromatography (HPLC) method for measurement of midazolam and its major metabolite, alpha-hydroxymidazolam, in clinical samples. Plasma or urine was mixed with 100 ng internal standard Ro 05 6669 and borate buffer, 0.1 M, pH 9. Midazolam and its related compounds were extracted into diethylether. The organic phase was evaporated to dryness. The residue was dissolved in HPLC mobile phase [methanol-isopropyl alcohol-perchloric acid, 0.5 microM (57:25:18)] and injected into the chromatograph. The separation of substances was performed on an Spherisorb S5CN 250 x 4.6 mm HPLC column maintained at 45 degrees C. The detection was performed by absorption measurement at 245 nm. At a flow rate of 1.7 ml/min, the retention times of Ro 05-6669, 1,4 dihydroxymidazolam, alpha-hydroxymidazolam, 4-hydroxymidazolam and midazolam were 4.0, 6.7, 7.8, 9.6, and 10.8 min, respectively. In the concentration range of 5 1,000 ng/ml, the calibration graphs for both compounds were linear. The coefficients of variation of the between-day and within-day assay were < 14% for the concentration range 5-10 and < 7% for the range 10-600 ng/ml. The limits of detection for midazolam and alpha-hydroxymidazolam were 2 and 4 ng/ml, respectively. This assay is more sensitive than earlier methods; it is simple and rapid, and it enables the quantification of midazolam and its alpha-hydroxy metabolite with very good precision and accuracy in human plasma and urine. PMID- 8236372 TI - Evaluation of the cloned enzyme donor immunoassay for measurement of phenytoin and phenobarbital in serum. AB - We report the evaluation of the cloned enzyme donor immunoassay (CEDIA) for the estimation of phenytoin and phenobarbital in serum. The assays were performed with a Hitachi 911 analyzer. Intra-assay coefficients of variation were from 1.5 to 4.4% for phenytoin and 1.6 to 5.5% for phenobarbital. Interassay coefficients of variation ranged from 1.8 to 5.3% for phenytoin and 2.9 to 4.8% for phenobarbital. Linearity was satisfactory, with a recovery of 103% at 11 mg/L and 110% at 22 mg/L phenytoin and 93% at 14 mg/L and 101% at 40 mg/L for phenobarbital. The detection limit was 1.2 mg/L for phenytoin and 0.6 mg/L for phenobarbital. Results of this assay correlated well with those of a conventional high-performance liquid chromatography (HPLC) method; r = 0.99 for phenytoin (n = 47) and r = 0.98 for phenobarbital (n = 48). The CEDIA was easy to handle and especially suitable for short turn-around time application. PMID- 8236373 TI - Identification of T-like and B-like lymphocyte subsets in sea star Asterias rubens by monoclonal antibodies to human leukocytes. AB - The axial organ of sea star Asterias rubens is a primitive immune organ. The total cell population was fractionated into two populations: adherent (B-like) and non-adherent cells (T-like) to nylon wool. These two cell subsets were previously defined as functionally acting as mammals T and B cells. In the present report, we pointed out that these T and B-like cells can be identified using mouse to human monoclonal antibodies. Reproducible results were obtained with anti-CD7 monoclonal antibody which detects the T-like cell subset and with anti-CD14 antibody that characterizes the B-like population. PMID- 8236374 TI - Involvement of IL-7 in the development of gamma delta T cells in the thymus. AB - The effect of IL-7 on the growth of thymic T lymphocytes was investigated by adding recombinant IL-7 into cell suspension cultures and submersion organ cultures (SOC) of murine fetal thymuses (FT) and newborn thymuses (NBT). FT and NBT were obtained from C57BL/6 mice at day 15 of gestational age and at day 3 after birth, respectively. In both cell suspension cultures and SOC, addition of IL-7 highly improved the cell recovery. In cell suspension cultures, addition of IL-7 resulted in the growth of gamma delta T-cells from FT-cells, whereas the same cytokine promoted the growth of both alpha beta and gamma delta T-cells from NBT-cells. These results may indicate that this cytokine is able to support the proliferation of T-cells of both alpha beta and gamma delta lineages. In marked contrast, in SOC, addition of IL-7 resulted in the growth of gamma delta T-cells not only in FT but also in NBT, despite the fact that the SOC of NBT without exogenous cytokine exclusively promoted the growth of alpha beta T-cells. A similar effect was also seen when IL-2 was added to NBT-SOC, though the skewing to gamma delta lineage was not so strong as in the case of IL-7. In addition, we found that IL-7 mRNA is expressed in the day 15 FT at a much higher level than in the adult thymus. These results strongly suggested that the production of a large amount of IL-7 synthesized in the FT is one of the major factors leading to the generation of gamma delta T-cells in FT. PMID- 8236375 TI - Thymic stroma-derived T-cell inhibitory factor (TSTIF) 1. TSTIF induces inhibition of antigen-stimulated T-cell proliferation. AB - The present study investigates the capacity of the MRL104.8a thymic stromal cell clone to modulate T-cell growth. The culture supernatant (SN) from the MRL104.8a stromal cell monolayer was added to cultures of Th-clones with or without T-cell receptor (TCR) stimulation as provided by antigen (Ag) plus splenic antigen presenting cells (APC). The results demonstrated that the MRL104.8a SN containing IL-7 activity induced dose-dependent proliferation of Th cells when they were not stimulated with Ag/APC. In contrast, addition of the same SN to cultures of Th cells during stimulation with Ag/APC resulted in potent dose-dependent inhibition of their proliferation. IL-7 contained in the SN was neither responsible for, nor involved in the inhibition event, because the inhibition was not observed with rIL-7 and was not neutralized by anti-IL-7 antibody. The growth inhibition of the Th clone in the presence of Ag plus APC was also induced by IL-10 or TGF-beta. However, the MRL104.8a SN-induced growth inhibition was mediated by a factor distinct from these cytokines, because (1) IL-10 cDNA was not amplified in polymerase chain reaction (PCR) products derived from MRL104.8a cells; (2) TGF beta cDNA was detected in the PCR products, but only marginal levels of TGF-beta activity in an active form were found in the MRL104.8a SN and the SN-induced inhibition was not prevented by anti-TGF-beta antibody; and (3) addition of rIL-7 to antigen-stimulated cultures containing rTGF-beta or rIL-10 induced IL-7 mediated Th proliferation, whereas the MRL104.8a SN-induced inhibition was still observed in the presence of excess rIL-7. Moreover, this factor, designated thymic stroma-derived T-cell inhibitory factor, was found to have a m.w. of 20-25 x 10(3) and to exhibit heparin-binding property. Thus, these results indicate that the MRL104.8a thymic stromal cell clone produces a potentially novel factor that induces inhibition of antigen-stimulated T-cell proliferation. PMID- 8236376 TI - Regulation of apoptosis in thymocytes. AB - Programmed cell death, or apoptosis, is tightly regulated during the development of T lymphocytes. Several studies have indicated that in normal mice, thymocyte are sensitive to apoptosis primarily during a brief period relatively late in the CD4+8+ maturation stage, when both positive and negative selection are thought to occur. One factor regulating sensitivity to apoptosis may be the expression and signalling capacity of the TcR/CD3 complex on developing thymocytes. In the present study, we report that sensitivity to apoptosis in immature thymocytes may also be regulated by a mechanism that can prevent induction of apoptosis in many thymocytes. This protective mechanism is induced by TcR/CD3 engagement and cross linking, as well as by agents that mimic TcR/CD3-dependent phosphoinositol bisphosphate hydrolysis and activate Ca++ fluxes and Protein Kinase C. Cyclosporin A (CsA) inhibits the protective mechanism, permitting the induction of apoptosis by TcR/CD3 or TcR/CD3-mimicking stimuli in otherwise resistant thymocytes. In contrast, mature naive T cells do not undergo apoptosis following stimulation by these agents, even in the presence of CsA, suggesting that in mature naive T-cells the apoptotic machinery itself is normally no longer inducible. We discuss the possible implications of these results for regulation of T-cell development. In this study we also demonstrate that CsA can inhibit the ability of accessory cells to trigger thymocyte apoptosis in accessory cell dependent assays, which may explain previous reports that CsA can inhibit the induction of thymic clonal deletion in vivo. PMID- 8236377 TI - Intestinal parasitic worms and the growth of children. PMID- 8236378 TI - Reappearance of Plasmodium malariae in Suriname? AB - Plasmodium malariae has not been reported from Suriname since 1979. In 1989 an increasing number of P. vivax infections among Bush-negroes returning from the eastern part of the interior was reported in Paramaribo. A microscopical re examination of all malaria cases in the eastern part of the country failed to confirm any P. vivax infections, but instead P. malariae infections were diagnosed. A study followed to determine the Duffy blood group antigens of 4 Bush negroes allegedly with a P. vivax infection in their medical history and of 28 and 32 unselected Bush-negroes and Amerindians respectively. Three of the 4 former Bush-negroes had the FyB antigen, while only 7% of the unselected Bush negroes had this antigen. This low frequency of the genotype is incompatible with reports of high P. vivax prevalences in Bush-negro populations. The Amerindians tested showed a low proportion of Fy0 genotype, which is compatible with the frequent diagnosis of P. vivax among this ethnic group. Reports of P. vivax infections among Bush-negroes are due to misdiagnosis of P. malariae, emphasizing the need to include all 4 species of human Plasmodium when (re)training microscopists. The question whether P. malariae reappeared in Suriname due to increased contact with the simian reservoir, or was simply missed, is discussed. PMID- 8236379 TI - Malaria attacks in children exposed to high transmission: who is protected? AB - In persons naturally exposed to high transmission, how significant, in terms of immune protection, is the occurrence or non-occurrence of a malaria attack during a given observation period? This question was studied in a West African village where Plasmodium falciparum malaria is holoendemic with intense and perennial transmission. A cohort of 94 children aged 4 months-14 years from Dielmo village, Senegal, was studied over 4 months, June-September 1990. 41 children had no malaria attack and 53 children suffered between one and 6 attacks. The average number of attacks was 1.80, 2.25, 1.87, 0.29 and 0.07, respectively, in children aged 4-11 months, 1-2, 3-6, 7-10 and 11-14 years. The transmission level was 75 infective bites per person. Analysis of the distribution of the number of attacks in individual children suggested that all children within the same age group had either the same, or a very similar, level of protection. This suggests that the acquisition of clinical protection in areas where malaria is highly endemic involves a progressive and homogeneous decrease of the probability of having a malaria attack (attacks occur less frequently as age increases, in all children), rather than the acquisition of complete protection by an increasing number of children. The differences between the number of clinical attacks observed in young children and their presumed exposure suggest that protective mechanisms become effective from the first reinfection onwards, and are independent of the cumulative exposure to a great variety of antigens. PMID- 8236380 TI - Evidence for a long-term increase in the incidence of Leishmania tropica in Aleppo, Syria. AB - We report the results of a short study of the epidemiology of Leishmania tropica in the Sheikh Maksoud District of Aleppo, Syria. The present and past status of infection in a community of about 100,000 people are assessed from prevalence and incidence data obtained by active and passive case detection, and from a skin test survey. L. tropica has apparently been endemic for at least 2-3 human generations in Aleppo, and incidence has increased over the past decade. The current estimated force of infection is 0.174/year, the incidence is about 5%, and the average age of infection is 14 years. L. tropica has the essential characteristics of a cyclic infectious disease, and the recent rise in incidence could be part of a long period cycle. A survey of leishmanial scars seriously underestimated the fraction of persons immune, as scar surveys usually do. We estimate that a passive case registration scheme, which has been established in response to growing concern about leishmaniasis in Aleppo, succeeds in recording and treating about one in 4 cases. PMID- 8236381 TI - Impact of the current National Bilharzia Control Programme on the epidemiology of schistosomiasis mansoni in an Egyptian village. AB - In a population-based study in the Nile Delta of Egypt, the modified Kato thick smear technique was used to study the epidemiology of Schistosoma mansoni. After 2 years of implementation of the National Bilharzia Control Programme by the Ministry of Health, the general prevalence dropped from 47.2% to 21.9%. The peak prevalence rate has shifted downwards in magnitude (40.4% compared to 72.1%) and backwards over the age scale (5-14 in contrast to 15-19 years age group). Redistribution of various infection grades has taken place. More than three quarters (75.6%) of the infected individuals have light infections in contrast to only one-fifth (20%) 2 years ago. The mean egg count for the entire sample population has dropped significantly (P < 0.05) from 128 to 17 eggs/g stools with no significant difference between males and females. This positive impact can be attributed largely to the case finding and treatment component of the programme. However, the incidence rate remained nearly the same before and 2 years after implementation of the programme (18.7 and 18.1%, respectively). The non-changing high incidence rate indicates that the knowledge, attitude and practice (KAP) of the population concerning water contact have not yet changed. Insufficient improvement in environmental sanitation may be a contributing factor. Continuation of serious efforts in case finding and treatment is recommended, with special emphasis on schoolchildren. This should be coupled with an intensive health education programme. PMID- 8236382 TI - Hydatid disease in the Hamar of Ethiopia: a public health problem for women. AB - A population-based study of hydatid disease was conducted among the Hamar of Ethiopia. Seven of 990 patients examined with a portable ultrasound scanner proved to have cysts (0.7% prevalence). Cysts were most common (4.7%) among women > or = 40 years of age. Only one male presented with a cyst. These results indicate a sex-specific hydatid public health problem in this tribe. PMID- 8236383 TI - The accumulation, distribution and viability of human parasitic nematode eggs in the sludge of a primary facultative waste stabilization pond. AB - Investigations into the enumeration, distribution and viability of nematode eggs in the sludge from a primary facultative waste stabilization pond showed that pond sludges are not nematologically safe for immediate land application. Up to 12% of Ascaris lumbricoides eggs were viable after the pond had been in operation for 2.5 years. Eggs were recovered from the sludge along the full length of the pond, although there were significant decreases in egg concentration and viability with increasing distance from the inlet. PMID- 8236384 TI - Initial report of a hepatitis investigation in rural Belize. AB - In spring 1991, Belizian health officials expressed concern about a possible hepatitis outbreak in a banana farming district. A study was designed to identify cases and to address the serological prevalence of hepatitis virus markers. Three populations were studied: (i) persons meeting a clinical case definition for hepatitis; (ii) designated banana workers; and (iii) people in a random sample of households in the community. Information was collected using questionnaires and sera were collected for laboratory testing. This report presents the preliminary results of a study conducted in June 1991. Among people who met the clinical case definition, 24% of 42 tested had immunoglobulin M antibody to hepatitis B virus (HBV) core antigen (anti-HBc IgM). In the worker and household survey populations, 284 and 280 people, respectively, were tested for anti-HBc IgM. In each group, 4% were positive. HBV surface antigen was found in 37% of 43 clinical cases, 18% of workers, and 13% of people in the household survey. Among the 3 study populations, the prevalence of HBV core antibody (anti-HBc) ranged from 73% to 81%. Almost all tested persons had evidence of prior hepatitis A virus infection. Evidence of prior infection with hepatitis viruses A and B was widespread, but an aetiology could not be established for most of the clinical cases. However, the prevalence of hepatitis B markers in this population was very high compared to other reports from the Caribbean. PMID- 8236385 TI - Hepatitis B in Ho Chi Minh City, Viet Nam. PMID- 8236386 TI - Prevalence of enteric viruses among hospital patients with AIDS in Kinshasa, Zaire. AB - Diarrhoea is the most common manifestation of acquired immunodeficiency syndrome (AIDS) in Africa. Numerous parasitic or bacterial agents have been implicated, but a pathogen-specific aetiology has not been found. Enteric viruses (i.e., rotavirus, small round structured viruses, coronavirus, and adenovirus) were detected by enzyme-linked immunosorbent assay or electron microscopy in faecal specimens of 17% of 198 consecutive adult admissions to a general medical ward of an urban hospital in Kinshasa, Zaire. Overall, 57% of patients were seropositive for infection with human immunodeficiency virus (HIV) 1; of these, 50% were classified as World Health Organization AIDS stage IV. The prevalence of enteric viruses in stool specimens did not differ significantly between patients with and without HIV infection, and was not associated with acute or chronic diarrhoea, or constitutional symptoms. However, a trend (P = 0.14) towards greater frequency of virus in stools from patients in the lower 3 quintiles of the CD4/CD8 T cell ratio was seen. This trend approached statistical significance (P = 0.07) with stratification by HIV infection. Although we found no evidence in this population to support a major pathogenic role for these viruses alone in the enteropathy of AIDS, increased viral shedding was weakly associated with immunodeficiency. PMID- 8236387 TI - Effect of a community-based Aedes aegypti control programme on mosquito larval production sites in El Progreso, Honduras. AB - This paper describes the results of a trial to assess the efficacy in reducing the density of larval production sites of utilizing community involvement in the planning and implementation of an Aedes aegypti control programme, in a dengue endemic city in Honduras. In addition to a substantial increase in knowledge about dengue transmission and prevention, a relative reduction in Ae. aegypti larval infestation indices was found in those city neighbourhoods where community involvement was utilized compared with their untreated counterparts. Several methods of improving the impact of this type of programme are discussed. PMID- 8236388 TI - Vibrio cholerae in South America: polymerase chain reaction and zymovar analysis. PMID- 8236389 TI - Polymerase chain reaction detection of Plasmodium falciparum in mosquitoes. AB - The polymerase chain reaction (PCR) procedure using a primer set derived from a repetitive deoxyribonucleic acid (DNA) sequence specific to Plasmodium falciparum was used to detect parasite DNA in mosquitoes. In laboratory-infected mosquitoes, PCR could detect as few as 10 sporozoites in a dissected salivary gland and a single oocyst in a dissected midgut. The ability to detect P. falciparum DNA in wild-caught mosquitoes indicated an advantage of the PCR over enzyme-linked immunosorbent assay for the detection of Plasmodium sporozoites in mosquitoes with low-grade parasite infections. PMID- 8236390 TI - Evaluation of the direct agglutination test as an immunodiagnostic tool for kala azar in India. AB - The direct agglutination test (DAT) has been assessed as a diagnostic procedure for visceral leishmaniasis. Fifty-six of 58 sera (96.5%) from confirmed cases of visceral leishmaniasis, whose bone marrow aspirates contained Leishmania donovani amastigotes, had agglutinating antibodies above the cut-off titre of 1:800. None of the sera from healthy control subjects from non-endemic or endemic areas had anti-leishmanial antibodies. Similarly, none of the sera obtained from cases of malaria or tuberculosis had agglutinating antibodies above the cut-off titre. A significant decline in agglutinating antibody titre in 3 cases following antileishmanial chemotherapy appeared to correlate with regression of clinical symptoms and the absence of amastigotes from bone marrow aspirates. One of 3 cases developed post-kala-azar dermal lesions and sera from this subject had an elevated agglutinating antibody titre. It is concluded that the DAT is a sensitive and specific test to confirm visceral leishmaniasis. As the formalin fixed promastigotes, stained with Coomassie blue, which are used as antigen could be stored at 4 degrees C for 6 months without any loss of ability to detect anti leishmanial antibodies, the DAT is recommended for use under field conditions. PMID- 8236391 TI - Dot-dye-immunoassay and dot-ELISA for the serological differentiation of acute and chronic schistosomiasis mansoni using keyhole limpet haemocyanin as antigen. AB - Two immunoassays, dot enzyme-linked immunosorbent assay (dot-ELISA) and dot-dye immunoassay (dot-DIA), using soluble egg antigen and keyhole limpet haemocyanin as antigens, were evaluated for the serological differentiation of 25 acute and 37 chronic patients infected with Schistosoma mansoni and 20 non-infected individuals, in comparison with ELISA. Efficiency was 92.7%, 90.0% for ELISA, dot ELISA and dot-DIA, respectively. Dipstick dot-ELISA and dot-DIA are described and shown to be reliable cheap and simple methods for the serological differentiation of acute and chronic schistosomiasis. PMID- 8236392 TI - A prospective study of the influence of alpha thalassaemia on morbidity from malaria and immune responses to defined Plasmodium falciparum antigens in Gambian children. AB - The protective effect of alpha thalassaemia (-alpha/alpha alpha) against morbidity from falciparum malaria was assessed in a prospective study of rural Gambian children. The gene frequency for single alpha-globin gene deletions was 0.12. Malariometric indices measured during cross-sectional surveys and morbidity from malaria determined by weekly surveillance were similar in children with alpha thalassaemia and in those with a normal alpha-globin genotype. However, the small number of children who carried both alpha thalassaemia and the sickle cell trait had fewer clinical episodes of malaria than children with the sickle cell trait alone. Specific antibody responses and cell-mediated immune responses in vitro to defined Plasmodium falciparum antigens were measured in children participating in the study. In general, there was no evidence of an increased prevalence or intensity of humoral or cell-mediated immune responses to the malaria antigens studied in children heterozygous for alpha thalassaemia compared with children with a normal alpha-globin genotype. PMID- 8236393 TI - Human leucocyte antigen (HLA) and malaria morbidity in a Gambian community. AB - Human leucocyte antigen (HLA) class I and class II typing was performed on 177 children in a rural area of The Gambia who were followed for 2 years in a longitudinal study of malaria morbidity. A comparison was made between those who experienced an episode of clinical malaria in one or both years and those who showed no evidence of infection in either year. No convincing association was found between morbidity and class I phenotype. An overall association of morbidity with the distribution of class II haplotypes was seen, but association with individual DR-DQ haplotypes were not conclusive. PMID- 8236394 TI - Coincidental severe Plasmodium falciparum infection and disseminated candidiasis. PMID- 8236395 TI - Schistosomiasis japonica on Jishan Island, Jiangxi Province, People's Republic of China: persistence of hepatic fibrosis after reduction of the prevalence of infection with age. AB - Hepatic fibrosis due to schistosomiasis japonica was examined by ultrasonography in a cross-sectional community study of 825 individuals on Jishan Island, Jiangxi Province, China. The prevalence of active infection was 39.4% with peak infection in the 10-19.9 years age group followed by a significant decline. A similar pattern was observed for intensity of infection. The prevalence of hepatomegaly in the midsternal line > or = 6 cm peaked at 60% in the fourth decade and remained elevated. A progressive increase in the severity of hepatic periportal fibrosis was observed with age, with advanced fibrosis peaking in the fifth decade. The proportion of individuals with advanced fibrosis was significantly greater in males than in females despite equivalent prevalence and intensity of schistosome infection. In addition, a positive association (P < 0.01) was found between periportal fibrosis and both hepatomegaly > or = 6 cm and splenomegaly. This study suggests that the natural history of schistosomiasis japonica in this hyperendemic community in China is marked by persistence of hepatomegaly and schistosome-induced periportal fibrosis in adults despite a decrease in the prevalence of infection. PMID- 8236396 TI - Crusted (Norwegian) scabies as a pre-diagnostic indicator for HTLV-1 infection. PMID- 8236397 TI - Fatal arteritis due to Pythium insidiosum infection in patients with thalassaemia. AB - Six thalassaemic patients had a distinct clinical syndrome characterized by progressive ischemia of the lower extremities, with ascending arteritis and thrombosis of the main arteries of the lower limbs. With periodic acid Schiff and Gomori's methenamine silver staining a large number of hyphae were revealed in the arterial wall and the outer part of the thrombus. Pythium insidiosum was isolated from 3 patients. The clinical course of the disease was progressive gangrene of the extremities and the patients invariably died when the infectious process reached the bifurcation of the aorta. There is no effective antimicrobial agent for the syndrome and radical amputation was the only method to ensure survival of the patients. P. insidiosum infection should be considered in thalassaemic patients with leg ulcers or arterial occlusion of the lower limbs. PMID- 8236398 TI - Comparison of intramuscular and intravenous quinine for the treatment of severe and complicated malaria in children. AB - To compare the efficacy and side effects of intramuscular (i.m.) and intravenous (i.v.) quinine, children in Mozambique with severe and complicated malaria between 6 months and 7 years were randomized to treatment with i.m. or i.v. quinine, both in a dosage of quinine dihydrochloride 20 mg/kg followed by 10 mg/kg every 8 h. Of 57 children treated with i.m. quinine, 4 died, 3 had neurological sequelae and 2 had sterile intramuscular abscesses. Of 47 children treated with i.v. quinine, 6 died and 1 had neurological sequelae. The mean parasite clearance time was 58.6 h in the i.m. group and 59.3 h in the i.v. group. Mean temperature clearance times were 56.1 and 51.8 h, and mean coma clearance times 40.4 and 38.7 h, respectively. None of these differences was statistically significant. Mean trough and peak concentrations of quinine were almost identical in the 2 groups, ranging from 10.5 to 12.6 mg/L, which is in the therapeutic non-toxic range. It is concluded that i.m. quinine is as effective as quinine by i.v. infusion in children with severe and complicated malaria; that minor local side effects can probably be avoided by using diluted quinine for i.m. injection; and that the optimal dose regimen for children with severe and complicated malaria in Africa at present is probably quinine salt 20 mg/kg followed by 10 mg/kg every 12 h. PMID- 8236399 TI - The potential of desipramine to reverse chloroquine resistance of Plasmodium falciparum is reduced by its binding to plasma protein. PMID- 8236400 TI - Efficacy of chloroquine-proguanil chemoprophylaxis against malaria in the Central African Republic. PMID- 8236401 TI - Spiramycin does not potentiate quinine treatment of falciparum malaria in pregnancy. PMID- 8236402 TI - The treatment of kala-azar in the Sudan with sodium stibogluconate: a randomized trial of three dosage regimens. AB - In a randomized study in the Sudan, 3 different regimens of sodium stibogluconate were compared in patients with parasitologically confirmed kala-azar (visceral leishmaniasis): 10 mg/kg for 30 d (38 patients), 20 mg/kg for 30 d (29 patients), and 20 mg/kg for 15 d (37 patients). Treatment failures were defined as death, partial response, relapse, or the development of post-kala-azar dermal leishmaniasis. The hazard ratio for failure of 20 mg/kg for 30 d vs. 10 mg/kg for 30 d 2.1 (95% confidence interval [CI] = 0.6, 7.6) and for 20 mg/kg for 15 d vs. 10 mg/kg for 30 d it was 1.7 (95% CI = 0.5, 6.1). No significant difference was detected between the 3 regimens in the rate of return to normal of haematological criteria, regression of spleen size, or weight gain. After 15 d treatment parasite clearance with 20 mg/kg for 30 d and 20 mg/kg for 15 d was more profound than with 10 mg/kg for 30 d (P < 0.05), but the difference was no longer present at the end of treatment. Further investigation of the effectiveness of short, intensive treatment regimens in the treatment of kala-azar is warranted. PMID- 8236403 TI - Treatment with liposomal amphotericin B of a child affected with drug-resistant visceral leishmaniasis. PMID- 8236404 TI - Circulating cathodic antigen levels in serum and urine of schistosomiasis patients before and after chemotherapy with praziquantel. PMID- 8236405 TI - Adverse reactions to ivermectin treatment for onchocerciasis. Results of a placebo-controlled, double-blind trial in Malawi. AB - A three-year placebo-controlled, double-blind trial involving 7148 persons was conducted to determine adverse reactions to ivermectin given annually for treatment of onchocerciasis by mass distribution. Musculoskeletal pains, oedema of the face or extremities, itching and papular rash were statistically associated with ivermectin treatment in the first year. In the second year of treatment these reactions were less frequently reported, although still more prevalent in ivermectin recipients. During the third year of treatment all persons received ivermectin. Persons who had missed ivermectin treatment in the second year did not have significantly more adverse reactions after treatment in the third year. Those who had received placebo during the first 2 years reported some adverse reactions more frequently following ivermectin than did those who had received ivermectin from the beginning of the study. Oedema was the adverse reaction of greatest concern to patients and this involved the face most commonly. A single episode of bullous skin lesions developed in 5 persons who had received ivermectin. These lesions did not reoccur with subsequent treatment. No episode of hypotension after treatment was observed. PMID- 8236406 TI - Absence of any adverse effect of inadvertent ivermectin treatment during pregnancy. PMID- 8236407 TI - Computer tomography of liver in alveolar echinococcosis treated with albendazole. AB - Fourteen cases of alveolar echinococcosis were examined by computed tomographic (CT) scanning of the liver, including 3 cases before treatment and 11 cases 2.0 7.5 years after long-term continuous albendazole therapy. In 7 cases the hepatic lesions were almost completely calcified and cured, while in 3 cases incomplete calcification of the peripheral margin of the hepatic lesions was observed. CT scanning of the one uncured case and the 3 cases before treatment showed active lesions with heterogeneous hypodense areas in the liver without calcification on the peripheral margin. Long-term continuous albendazole therapy can result in parasitological cure with death of the parasite. PMID- 8236408 TI - Detection of sporozoites of Plasmodium vivax and Plasmodium falciparum in mosquitoes by ELISA: false positivity associated with bovine and swine blood. AB - Blood samples from cows and pigs were tested for possible cross-reactivity with a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) kit designed for detection of human malaria sporozoites in mosquitoes. The results revealed that 4 of 16 cows (25%) reacted positively with both Plasmodium falciparum (2A10) and P. vivax (NSV3) monoclonal antibodies and 8 (50%) were positive with NSV3 only. One of 12 pigs (8.33%) was positive with both antibodies, and 2 (16.6%) were positive with NSV3 only. The positivity was associated with plasma, but not with the blood cell fraction. Antigenic extracts of Sarcocystis, Toxoplasma gondii and Trypanosoma evansi gave negative ELISA results, suggesting that these were not the factors in animal blood which gave positive results. Laboratory Anopheles dirus A fed on blood of a positive cow by membrane feeding also gave a positive ELISA result. Furthermore, some blood-fed culicine mosquitoes collected directly from a positive cow were ELISA-positive. The cross-reactive factor(s) in plasma has (have) not yet been identified. These false positive ELISA results could complicate the assessment of sporozoite rate in mosquito populations if the study were carried out by ELISA only, especially in areas where cattle and swine are present. PMID- 8236409 TI - Metacyclogenesis of Leishmania (Viannia) braziliensis in vitro: evidence that lentil lectin is a marker of complement resistance and enhanced infectivity. AB - Axenic culture in modified Grace's medium was used to induce metacyclogenesis of Leishmania (Viannia) braziliensis in vitro. Morphological characteristics, lectin agglutination profiles, susceptibility to complement lysis, and infectivity in vivo were compared between metacyclic promastigotes and promastigotes in mid-log phase growth. Short, arrow-like promastigotes and round, oval promastigotes were defined as putative metacyclic forms on the basis of being highly motile and free swimming, with a small cell body and long flagellum. These forms increased during metacyclogenesis to > 80% whereas long-bodied, slender promastigotes and intermediate slender promastigotes declined progressively. Lentil lectin selectively agglutinated L. braziliensis after the induction of metacyclogenesis, whereas concanavalin A, wheat germ agglutinin and peanut agglutinin similarly agglutinated metacyclic promastigotes and mid-log phase promastigotes. Metacyclic promastigotes survived in 7.5%-20% human serum whereas mid-log phase promastigotes did not. Five hundred metacyclic promastigotes were highly infective to hamsters whereas 500 mid-log phase promastigotes rarely caused any lesion. Specific agglutination by lentil lectin should allow purification of metacyclic organisms for standardization of immunoprotection and challenge experiments. PMID- 8236410 TI - Determination of anti-Cryptosporidium coproantibodies by time-resolved immunofluorometric assay. AB - The role of the mucosal immune response against Cryptosporidium has been suggested by studies on the therapeutic effects of hyperimmune colostrum. In order to study the intestinal response to this infection, we have developed a sandwich-type time-resolved immunofluorometric assay for the determination of anti-Cryptosporidium coproantibodies. This assay has the inherent sensitivity of an immunoassay without the problems due to background responses from other biological compounds, and is thus suitable for faecal samples. The intra-assay coefficients of variation (5.1%, 4.6%, and 5.8% for immunoglobulins (Ig) A, M and G respectively), inter-assay coefficients of variation (9.4%; 10.5% and 12.2% for IgA, IgM and IgG, respectively) and specificity (100% for all 3 isotypes) were all satisfactory. Using this assay to study 12 patients with the acquired immune deficiency syndrome (AIDS) who were infected with cryptosporidiosis, we found a marked elevation of anti-Cryptosporidium IgA and IgM coproantibody titres relative to 18 healthy control values, but no correlation with the gravity of the infection in terms of oocyst shedding. These results suggest that a non protective mucosal immune response develops to Cryptosporidium in AIDS patients. PMID- 8236411 TI - Cost-effectiveness of hepatitis B vaccine in The Gambia. AB - Hepatitis B vaccine has been progressively introduced into the Gambian programme of immunization since 1986. Other African countries are considering introduction but require evidence of cost-effectiveness to justify such a decision. The cost of the Expanded Programme on Immunization in The Gambia, which includes hepatitis B vaccine, was calculated for 1988. Estimates of the effects of this programme on the incidence of liver cancer were made, based on the national cancer registry and a case control study of primary liver cancer and hepatitis B, regarding hepatitis B vaccine costs as incremental. This gave an estimate of the cost of averting a death from liver cancer in the range US$150-200 (assuming a vaccine cost of US$1 per dose). This indicates that universal hepatitis B immunization is comparable, in terms of cost-effectiveness, to other health interventions in less developed countries. PMID- 8236412 TI - Yellow fever vaccination and pregnancy: a four-year prospective study. AB - During an outbreak of yellow fever (YF) in Nigeria in 1986-1987, women at various stages of pregnancy were vaccinated against YF, either because those pregnancies were not known at the time or because they requested vaccination out of fear of acquiring the disease. This offered an opportunity to assess the safety and efficacy of YF vaccine in pregnant women and the effect of this vaccine on their newborn children. Pre-vaccination and post-vaccination serum samples from the vaccinated pregnant women were tested by enzyme-linked immunosorbent assay and by neutralization tests for antibody to YF virus. The results showed that the antibody responses of these pregnant women were much lower than those of YF vaccinated, non-pregnant women in a comparable control group. Follow-up of these women and their newborn children for 3-4 years showed no abnormal effect that could be attributed to the YF vaccine, which suggests that vaccination of pregnant women, particularly during a YF epidemic, may not be contraindicated. PMID- 8236413 TI - Tuberculosis, AIDS and chemoprophylaxis. PMID- 8236414 TI - Hydatid immunoblot test and cross-reactivity with sera from patients with cysticercosis. PMID- 8236415 TI - Strongyloidiasis on the Thai-Cambodian border. PMID- 8236416 TI - It seemed a pity to throw away the red cells: selective component collection. PMID- 8236417 TI - Hemophilia: continued challenges but reason for optimism on many fronts. PMID- 8236418 TI - A prospective study to identify the risk factors associated with acute reactions to platelet and red cell transfusions. AB - It is generally assumed that febrile nonhemolytic transfusion reactions are an immunologically mediated reaction involving the recipient's plasma and the white cells in the donor unit. This has led to the use of white cell reduction and pretransfusion medication, to try to minimize these reactions. To better understand febrile transfusion reactions, a prospective study was performed in which all patients receiving platelet and red cell transfusions in a tertiary care medical center were interviewed before and after transfusion to obtain information about the typical presentation of the syndrome. It was found that transfusion reactions were much more frequently associated with platelet transfusion (30.8%) than with red cell transfusion (6.8%, p < 0.0005). The routine use of antipyretics prevented most episodes of fever but did not prevent the occurrence of other symptoms such as chills, cold, and discomfort. The application of logistic regression analysis revealed that the dominant factor determining the risk of a reaction was not white cell contamination, but the age of the component (p < 0.005). The significant relationship between reaction and the increasing age of the component suggests that cytokines released in the component during storage may be responsible for many reactions to blood components. PMID- 8236419 TI - Experimental animal model of refractoriness to donor platelets: the effect of plasma removal and the extent of white cell reduction on allogeneic alloimmunization. AB - Platelet transfusion effectiveness may be limited in multiply transfused patients by the development of the refractory state. White cell (WBC)-reduction filters with variable efficiency (1-3 log10 reduction) are available and have been shown to be effective in reducing the incidence of platelet alloimmunization. However, the threshold number of WBCs below which alloimmunization would no longer occur is yet to be determined. A previously established animal model was used to examine the relative efficiency of second- and third-generation filters in reducing the frequency of refractoriness to allogeneic platelets. In this model, California Black rabbits are used as blood donors and New Zealand White rabbits as transfusion recipients. Eight weekly transfusions of either second-generation or third-generation WBC-reduced blood resulted in no difference between the two groups in mean platelet survival and rate of refractoriness to allogeneic platelets. To evaluate the possible incremental benefit of removing supernatant plasma to prevent platelet refractoriness, experiments were performed in which groups of animals were given transfusion(s) with red cell suspensions that had been WBC-reduced or both plasma-depleted and WBC-reduced. A significantly lower rate of allogeneic platelet refractoriness was seen in the rabbits that received WBC-reduced and plasma-depleted red cells than in those that received red cells that had been WBC-reduced only. These data provide evidence that the combined use of plasma depletion and WBC reduction can decrease still further the frequency of refractoriness produced by allogeneic blood transfusions. PMID- 8236420 TI - Transfusion-associated Serratia marcescens infection: studies of the mechanism of action. AB - The growth of two strains of Serratia marcescens in blood components was tested in this study. One of the strains had been implicated in the epidemic of transfusion-associated sepsis experienced in Denmark and Sweden in 1991. In whole blood with a final concentration of 100 colony-forming units per mL of S. marcescens, there was an immediate reduction of more than 95 percent of colony forming units, but no reduction of the bacterial concentration if the blood had been white cell-reduced before inoculation. This is interpreted as an effect due to phagocytosis by white cells and as a lack of bactericidal effect of the plasma. A reduction to 10 percent of the original concentration, observed if the blood had a nominal content of white cells, was most likely due to phagocytosis. White cell reduction by filtration after inoculation further reduced the bacterial concentration of one of the strains tested, but, after a 1-week lag phase, growth accelerated to high concentrations by 6 weeks. In platelet-rich plasma prepared from S. marcescens-inoculated units, abundant growth was found after 24 hours, increasing to very high concentrations (10(12) colony-forming units/mL) during 10-day storage at 22 +/- 2 degrees C. Keeping the whole blood at ambient temperature for 20 hours before preparation of platelet-rich plasma caused only temporary reduction of bacterial concentration in the S. marcescens experiments, but resulted in a complete absence of bacteria in the platelet-rich plasma for 10 days in control experiments performed with Staphylococcus epidermidis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236421 TI - Prospective study of the evaluation of hepatitis C virus infectivity in a high purity, solvent/detergent-treated factor VIII concentrate: parallel evaluation of other markers for lipid-enveloped and non-lipid-enveloped viruses. The Ad Hoc Study Group of the Fondazione dell'Emofilia. AB - This prospective study was carried out with the aim of evaluating the efficacy of solvent/detergent inactivation of the hepatitis C virus (HCV) as applied to a chromatographic factor VIII concentrate. In parallel, the markers for other viruses, either lipid-enveloped (human immunodeficiency virus types 1 and 2 [HIV 1 and -2] and hepatitis B virus [HBV]) or non-lipid-enveloped viruses (such as B19 parvovirus and hepatitis A virus [HAV]) were evaluated. The study included 14 hemophilia centers, which enrolled 36 previously untreated patients (median age, 3 years; range, 1-56). The length of follow-up was 12 months, during which HCV (first- and second-generation assays and recombinant immunoblot assay), HIV-1 and -2, HBV, HAV (IgG and IgM), and parvovirus (IgG and IgM) antibodies, as well as alanine aminotransferase values were evaluated. Thirty-one patients were analyzable; none seroconverted for HCV, HBV, or HIV after exposure to a total of 165,000 IU of factor VIII (41 different lots). In one patient, alanine aminotransferase values rose to 167 mU per mL, 6 weeks after the first concentrate infusion, and this patient seroconverted for HAV 1 week later. Furthermore, 10 patients seroconverted for parvovirus during follow-up. This study suggests that the solvent/detergent method of virus inactivation is efficient in relation to lipid-enveloped blood-borne viruses but not in relation to non-lipid-enveloped viruses. PMID- 8236422 TI - Red cell collection by apheresis technology. AB - To determine the feasibility of collecting 2 units (450 mL) of red cells per donation by apheresis technology, apheresis red cell collections were compared to whole-blood donations. Forty blood donors were equally divided between the two study arms on the basis of gender and iron supplementation (650 mg ferrous gluconate/day vs. no supplementation). During the 1-year study period, the apheresis participants donated 450 mL of red cells three times, and the whole blood donors gave 225 mL of red cells (1 unit of blood) on six occasions. There were no reported side effects during the 102 whole-blood donations, whereas symptoms were noted in 83 percent of the 59 apheresis procedures. The most common symptoms were numbness and tingling, which were relieved by a decrease in the plasma-return rate or by the administration of oral calcium supplements. Seven donors dropped out or were deferred during the study. Two whole-blood donors left with medical problems unrelated to the study, one apheresis donor and one whole blood donor dropped out of the study because of excessive fatigue, and three non iron-supplemented whole-blood donors had unacceptably low hematocrit levels. By the end of the study, 70 percent of the apheresis donors considered the procedure acceptable, 15 percent were undecided, and 15 percent thought it was not acceptable. As measures of iron balance, the serum ferritin and the red cell zinc protoporphyrin:heme ratios were significantly more abnormal in the non-iron supplemented donors than in the iron-supplemented donors. However, there were no differences in iron balance according to the donation method. PMID- 8236423 TI - The efficacy of subcutaneous recombinant human erythropoietin in the correction of phlebotomy-induced anemia in autologous blood donors. AB - The efficacy of subcutaneous recombinant human erythropoietin (rhEPO) (500 U/kg; administered twice a week during the 3 weeks before surgery) in the recovery of preoperative hemoglobin concentrations within a 3-week period was studied in 40 patients, each of whom donated 2 units (900 mL) of blood for their own use before total hip replacement surgery. Twenty autologous blood donors received rhEPO (EPO group) and 20 were not treated (control group). The initial hemoglobin concentration (14.0 +/- 1.0 g/dL [140 +/- 10 g/L]) was completely recovered before surgery (14.0 +/- 1.6 g/dL [140 +/- 16 g/L]) in the EPO group, while a decrease from 13.8 +/- 1.1 to 12.2 +/- 1.3 g per dL (138 +/- 11 to 122 +/- 13 g/L) was observed in the control group. The preoperative reticulocyte count showed more than sixfold increase in the EPO group, whereas a twofold to threefold increase was found in the control group. Serum ferritin concentration fell to 42 +/- 29 micrograms per L in the EPO group and to 54 +/- 35 micrograms per L in the control group. The postoperative serum erythropoietin concentration in the EPO group was significantly lower than that in the control group, but it did not differ from the pretreatment value and was attended by a higher hemoglobin concentration after surgery. Only transient flu-like symptoms were mentioned by patients who were treated with rhEPO. Changes in blood pressure or platelet count or other adverse events were not observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236424 TI - Chagas' disease diagnosis: evaluation of several tests in blood bank screening. AB - Blood transfusion is one of the principal routes of transmission of Chagas' disease, a major endemic disease in Latin America. Methods for blood screening are not accurate and may yield false results that lead to high social and economic costs. This study compares two methods of diagnosing Chagas' disease (indirect immunofluorescence and hemagglutination) and several enzyme-linked immunosorbent assays (ELISAs) with regard to specificity and sensitivity, by using human sera with known serologic and parasitologic characteristics, as well as samples with discrepant results on conventional serologic tests. An ELISA using recombinant antigens showed no cross-reactivity with sera that were positive for other diseases. All evaluated ELISAs performed well, and their use may lead to a reduction of more than 50 percent in the number of discordant sera. Further improvements are needed in view of the complexity of the serologic diagnosis of Chagas' disease. PMID- 8236425 TI - Comparison of plateletpheresis with a standard and an improved collection device. AB - A blood cell separator with a specialized separation chamber ([TNX-6]CS-3000 Plus) was developed for the collection of platelet concentrates with higher platelet yields and lower white cell contamination than obtained with the standard blood cell separator (CS-3000). To compare these devices, normal donors were scheduled for paired plateletpheresis procedures spaced 4 weeks apart, with one procedure using the CS-3000 Plus and the other using the CS-3000. Overall, the platelet yield per unit (mean +/- SEM) was 4.3 +/- 0.1 x 10(11) with the CS 3000 Plus versus 3.7 +/- 0.1 x 10(11) with the CS-3000 (p < 0.001), and the white cell contamination per unit (mean +/- SEM) with the former was 2.4 +/- 0.7 x 10(6) versus 84.1 +/- 21.1 x 10(6) with the latter (p < 0.001). The sequence of procedures (i.e., the order in which the devices were paired) was selected randomly, and similar results were found regardless of sequence. When donors with predonation platelet counts of > or = 200 x 10(9) per L (n = 21) were studied separately, 76 percent of the collections by the CS-3000 Plus contained > or = 4 x 10(11) platelets versus 34 percent of those by the CS-3000 (p < 0.01), and 93 percent of the collections by the former contained < 5 x 10(6) white cells (69% contained < 1 x 10(6)) versus 0 percent of those by the latter (p < 0.01). Thus, platelet collections with the TNX-6 chamber consistently demonstrated high platelet yields and strikingly low white cell contamination--qualities that justify converting standard devices to devices with a TNX-6 chamber. PMID- 8236426 TI - A comparison of polymerase chain reaction and an infectivity assay for human immunodeficiency virus type 1 titration during virus inactivation of blood components. AB - Three examples of human plasma-derived concentrates, intermediate-purity factors VIII and IX, and fibrinogen were spiked with tissue culture-grown human immunodeficiency virus type 1 (HIV-1) strain RF. All examples were freeze-dried and heated at 80 degrees C for 72 hours by using validated production process models. HIV-1 infectivity was measured by a syncytial infectivity assay in C8166 cells and then compared with levels determined by nested HIV polymerase chain reaction (PCR). The infectivity assay demonstrated a reduction index of at least 4.5 log10, while PCR showed an average 1.7 log10. Large amounts of HIV-1 RNA (10(5)) were still detectable by PCR in samples in which infectivity assays failed to detect any HIV-1. These data suggest that HIV-1 PCR levels do not parallel HIV-1 infectivity levels during virus-inactivation procedures involved in coagulation factor concentrate production. PCR was able to detect the RNA associated with inactivated HIV-1 particles in the factor concentrates, which allows the conclusion that PCR is not a useful test with which to monitor virus inactivation procedures such as heating at 80 degrees C for 72 hours. This judgment contrasts with the more definite and sensitive role of PCR in diagnosing HIV-1 infection in patients in whom a positive HIV-1 PCR result correlates with active HIV-1 infection and with PCR's usefulness in monitoring virus removal. PMID- 8236427 TI - Analysis of a linear peristaltic infusion device for the transfusion of red cells to pediatric patients. AB - A linear peristaltic infusion device was evaluated for red cell (RBC) transfusion in the pediatric and neonatal setting. CPDA-1 RBC units (n = 24) divided into six groups of 4 units each underwent simulated transfusion. Blood was infused by using manufacturer-provided administration sets with either a 21-gauge needle or a 24-gauge catheter. Filters were used in two groups to evaluate the effect of negative pressure on filter function. Two groups of RBCs less than 1 week old were washed, irradiated, and infused at 5 mL per hour, by using a standard administration set, or at 10 mL per hour, by using a syringe set. Four-week-old RBCs (washed and irradiated, irradiated and filtered, filtered only, or unmanipulated) were infused at 100 mL per hour. Paired samples from 0 and 2 hours before and after infusion were analyzed for hemoglobin, hematocrit, RBC count, plasma hemoglobin, lactate dehydrogenase, potassium, alanine aminotransferase, and aspartate aminotransferase. Hausser and Nageotte hemocytometers were used to perform white cell (WBC) counts when a filter was used. By analysis of variance and percentage of change, data from 0 and 2 hours before and after infusion were compared. No clinically or statistically significant differences were seen for hemoglobin, hematocrit, or RBC count. The difference in preinfusion and postinfusion plasma hemoglobin levels in washed RBCs at 2 hours was statistically but not clinically significant (14.5 +/- 6.8 vs. 19.3 +/- 7.1 mg/dL). No clinically significant differences were noted for the remaining analytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236428 TI - Autologous and allogeneic red cell survival studies in the presence of autoanti AnWj. AB - Chromium survival studies were performed with AnWj-positive allogeneic blood in a patient with autoanti-AnWj. 99mTc-labeled autologous RBCs that had depressed AnWj expression had normal survival (77% [94.7% 51Cr equivalent]) at 24 hours, whereas 51Cr-labeled allogeneic AnWj-positive cells had 76 percent survival at 24 hours and 55 percent survival at 7 days. These studies suggest that the specificity of the autoantibody may have implications for transfusion therapy when the development of such autoantibodies is associated with decreased antigen expression on the patient's cells. PMID- 8236429 TI - An anti-EnaTS detected in the serum of an MiI homozygote. AB - The serum of EH reacted with all red cells (RBCs) except her own, ficin- or trypsin-treated red cells, and En(a-) red cells. This reactivity defined an anti EnaTS specificity. The red cells of the proposita typed as M-N+S-S+, Vw+Mur-Hil Hut-Anek-Lane-, Wr(a-b+), EnaKT+. Red cells of five relatives were Vw+ and positive with her serum. Titration studies suggest that EH is genetically an MiI homozygote and that her Vw+ relatives are MiI heterozygotes. There is no history of consanguinity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting studies have agreed with the serologic observations. A variant sialoglycoprotein of faster mobility than normal glycoprotein A, but no normal glycoprotein A, was detected on her red cells. Treatment with N-glycanase did not alter the mobility, which indicated that there was no N-glycosylation of residue 26. These findings are in agreement with the reported properties of the Mi.I specific glycoprotein A. The relatives' Vw+ red cells showed the variant sialoglycoprotein and normal glycoprotein A. EH appears to be the first reported MiI homozygote. PMID- 8236431 TI - National Hemophilia Foundation 44th Annual Meeting. November 19-22, 1992. Abstracts. PMID- 8236430 TI - The molecular basis for paroxysmal nocturnal hemoglobinuria. PMID- 8236432 TI - The effect of blood transfusion on serum ferritin level. PMID- 8236433 TI - Enzyme techniques in pretransfusion testing. PMID- 8236434 TI - Beware the Jabberwock and other perils of readability formulae. PMID- 8236435 TI - How receptor tyrosine kinases activate Ras. PMID- 8236436 TI - Does protein kinase C play a role in rhodopsin desensitization? PMID- 8236437 TI - Why NADP? PMID- 8236438 TI - Use of genetically prepared enzyme conjugates in enzyme immunoassay. AB - Enzyme immunoassay, using enzymes crosslinked to either antibodies or antigens, has proved a valuable immunological tool for many years. Recently, gene fusion techniques have been used to prepare these enzyme conjugates. This method may be especially advantageous in cases where (1) the antigen is difficult and costly to obtain in large quantities or (2) when the activity of the marker enzyme or the affinity of the antibody or antigen is severely reduced or even destroyed by use of conventional linking methods such as chemical crosslinking. This article uses specific examples to illustrate the potential of gene fusion as a conjugation method. PMID- 8236439 TI - New developments in the study of biomolecular associations via sedimentation equilibrium. AB - The measurement and analysis of sedimentation equilibrium provide one of the most powerful techniques for quantitative characterization of reversible and irreversible macromolecular associations in solution. The use of this technique by nonspecialists has been greatly helped in recent years by the development of new instrumentation, new types of experiments and new PC-based software for computer-aided analysis of experimental results. PMID- 8236440 TI - The pH dependence of the apparent equilibrium constant, K', of a biochemical reaction. AB - Biochemical reactions can be discussed in terms of chemical equations or biochemical equations. Both are right and both are needed. Therefore, it is important to be able to distinguish between them at a glance. Sometimes the relations between the apparent equilibrium constant 'K" for a biochemical reaction and the equilibrium constant, K, for a reference reaction written as a chemical equation is simple, and sometimes it is complicated. This article will discuss several examples. PMID- 8236441 TI - Transcriptional activator components and poxvirus DNA-dependent ATPases comprise a single family. PMID- 8236442 TI - Sequence similarity of phospholipase A2 activating protein and the G protein beta subunits: a new concept of effector protein activation in signal transduction? PMID- 8236443 TI - Translational introns: an additional regulatory element in gene expression. AB - The linear expression of a gene can be interrupted by the well-known RNA introns and the recently discovered protein introns. In both cases, splicing mechanisms physically excise the unexpressed segments. In this article we describe a third category of introns that we call 'translational introns'. These functional introns are not excised through a splicing mechanism; instead, the translational machinery bypasses a segment of the coding sequence of an mRNA. We suggest that 'translational introns' are part of a regulatory mechanism that may sense changes in the rate of translation and thereby control the ratio of alternative gene products. PMID- 8236444 TI - Bacterial L-serine dehydratases: a new family of enzymes containing iron-sulfur clusters. AB - Two families of enzymes are described which catalyse identical chemical reactions but differ in their prosthetic groups and hence in their mechanism of action. One family, the pyridoxal-5'-phosphate (PLP)-dependent L-threonine dehydratases, also use L-serine as substrate. The other, hitherto unrecognized family is the iron dependent, highly specific bacterial L-serine dehydratases. It has been shown that L-serine dehydratase from the anaerobic bacterium Peptostreptococcus asaccharolyticus contains an iron-sulfur cluster but no PLP. A mechanism for the dehydration of L-serine which is similar, but not identical, to that of the dehydration of citrate catalysed by aconitase is proposed. PMID- 8236445 TI - Lens crystallins: gene recruitment and evolutionary dynamism. AB - In a novel evolutionary process, enzymes and stress proteins have undergone direct gene recruitment as eye lens crystallins in a number of independent events. This may have allowed a dynamic response to changing visual environment during evolution. In spite of their diversity, many crystallins may share an origin in essential developmental processes such as cell elongation. PMID- 8236446 TI - The control of apoptosis in mammalian cells. AB - This article summarizes what is now known, more than 100 years after this prescient observation, of the mechanisms by which death of mammalian cells is regulated. Particular attention is paid to the elimination of self-reactive thymocytes, the induction of cell death during cancer therapy and the mechanism(s) by which cells die following growth factor removal. PMID- 8236447 TI - The BIOSCI newsgroups--computer networks changing biology. PMID- 8236448 TI - On a (beta-) roll. PMID- 8236449 TI - G proteins and beta ARK: a new twist for the coiled coil. PMID- 8236450 TI - Straightening out the dihedral angles. PMID- 8236451 TI - Nano-manipulation of actomyosin molecular motors in vitro: a new working principle. AB - Techniques have been recently developed that allow the direct observation of single actin filaments and their manipulation, using glass microneedles, in the nanometer range. Further development of these techniques has made possible the detection of subpiconewton-level forces of individual myosin heads. This in vitro motility model is sensitive in the submillisecond range and has allowed us to determine the force generation of an actomyosin motor directly at the molecular level. The results have led to a new conceptual framework for chemo-mechanical energy transduction in the molecular motor. PMID- 8236452 TI - How many intronic snRNAs? PMID- 8236453 TI - The PH domain: a common piece in the structural patchwork of signalling proteins. AB - The 'pleckstrin homology' domain is an approximately 100-residue protein module that has recently been added to the domain catalogue of signalling proteins. For this review we have made an extensive database search using a profile search method, and found a number of additional proteins that may contain PH domains. The PH domain is present in many kinases, isoforms of phospholipase C, GTPases, GTPase-activating proteins and nucleotide-exchange factors, including such proteins as Vav, Dbl and Bcr, and there are two PH domains in a guanine nucleotide releasing factor of Ras. Many PH-domain-containing proteins interact with GTP-binding proteins. We have also identified a PH domain in beta-adrenergic receptor kinase exactly in the region that has already been shown to be involved in binding to the beta and gamma subunits of a heterotrimeric G protein. This suggests that PH domains may be involved in interactions with GTP-binding proteins. PMID- 8236454 TI - Inhibitors of Ras farnesyltransferases. AB - Farnesyltransferase catalyses the post-translational modification of proteins by a cholesterol precursor, farnesylpyrophosphate. One of the substrates of this enzyme is the product of the ras oncogene. Recently, inhibitors of farnesyltransferase have been identified through two different approaches: microbial screens for natural compounds, and substrate analogues. These inhibitors may be useful in blocking the action of Ras proteins, in further characterizing protein prenyltransferases, and in elucidating the regulation of cholesterol metabolism. PMID- 8236455 TI - Brushing up on bristles: complex genes and morphogenesis in molds. PMID- 8236456 TI - Use of glycerol for enhanced efficiency and specificity of PCR amplification. PMID- 8236457 TI - Interpreting sequence motifs. PMID- 8236458 TI - Assembly and functions of the spindle pole body in budding yeast. AB - The spindle pole body (SPB) serves as the centrosome in yeasts and in a variety of other lower eukaryotes. In Saccharomyces cerevisiae, this organelle controls the assembly of all microtubules in the cell, acting not only as a pole of the mitotic or meiotic spindle but also as the site from which cytoplasmic microtubules emanate. The distinctive structure of the SPB has permitted definition of discrete stages in its duplication and behavior at all stages of the yeast life cycle. In association with genetic analyses, studies of the yeast SPB are providing insights into the mechanisms that control centrosomal behavior in this model eukaryote. PMID- 8236459 TI - Developmental regulation of human beta-globin gene transcription: a switch of loyalties? AB - Synthesis of different hemoglobin polypeptides during the early stages of human development is principally regulated by transcriptional control mechanisms that determine which of the five beta-type globin genes is expressed. The means by which this is achieved have been scrutinized for several decades, and insights have been gained from introducing segments of the human beta-globin locus into transgenic mice, and from analysis of naturally occurring mutations at the locus. I describe here a model which attempts to resolve several of the current puzzles and provides simple, testable predictions for how differential beta-globin gene transcription might be achieved during human development. PMID- 8236460 TI - Requiem for distributive segregation: achiasmate segregation in Drosophila females. AB - The segregation of achiasmate chromosome pairs at meiosis I is not brought about by a single 'distributive system' as previously thought, but rather by two separate mechanisms. One system uses the pairing of proximal heterochromatic sequences to mediate the segregation of achiasmate homologs-an observation that, at long last, defines a function for heterochromatin. The other system facilitates the segregation of heterologous chromosomes, by an as yet undiscovered mechanism. PMID- 8236461 TI - From cadherins to catenins: cytoplasmic protein interactions and regulation of cell adhesion. AB - Classical cadherins are complexed via their cytoplasmic domains with alpha-, beta and gamma-catenin. This complex formation links cadherins to the actin filament network and to other transmembrane and cytoplasmic proteins. alpha-Catenin is homologous to vinculin, and beta-catenin to the product of the Drosophila gene armadillo, while gamma-catenin seems to be identical to plakoglobin. Catenins are part of a higher order protein structure that is of crucial importance for the adhesive function of cadherins. A working model of the construction and regulation of this multiprotein interaction is proposed. PMID- 8236462 TI - In vivo mutation at the human HPRT locus. AB - The molecular nature of mutations that arise in vivo at the human hypoxanthine guanine phosphoribosyltransferase (HPRT) locus can be determined. A wide variety of such mutations can be detected, including large and small deletions, frameshift mutations and single-base substitutions, as well as alterations that cause aberrant mRNA splicing. Here, we review the available information on mutations at this locus. PMID- 8236463 TI - Urgent need for a new approach to the diagnosis of tuberculosis in developing countries in the decade of AIDS. AB - In many developing countries with a high prevalence of both tuberculosis and HIV infection a dramatic increase of tuberculosis is seen. Our present ability to diagnose tuberculosis in developing countries has its limitations. This review article discusses old and new approaches to the diagnosis of tuberculosis and their value for developing countries. PMID- 8236464 TI - Malaria chemosuppression during pregnancy. IV. Its effects on the newborn's passive malaria immunity. AB - The effect of malaria prophylaxis during pregnancy on the levels of cord blood anti-sporozoite antibodies was investigated in 203 newborns in Muheza, Tanzania. Mothers of 76 newborns had received prophylaxis with proguanil daily (PROG), 66 chloroquine once weekly (CQ), and 61 got a combination of the two drugs (CQ+PROG). Prophylaxis with PROG or CQ+PROG was more efficacious than with CQ. The mean antibody titres were comparable in all three groups, despite titres being significantly low in mothers of the CQ+PROG group. In 93% of 167 paired maternal-cord sera, maternal titres were higher than cord titres. The correlation between maternal and cord titres was low. Parity, placental malaria, and baby maturity showed little influence on titres. Titres of babies delivered by Caesarean section or whose placenta weighed between 0.75 and 1 kg were comparatively low. The first occurrence of a malaria parasitaemia in infants was independent of the levels of cord titres at birth. The results suggested that chemoprophylaxis as effective as PROG or CQ+PROG in holoendemic areas, insignificantly affects maternal-foetal transfer of anti-sporozoite antibodies, and that levels of these antibodies at birth do not modulate the first occurrence of infancy malaria parasitaemia. Interference with the maternal-foetal transfer of this antibody and possibly other component antibodies of passive immunity should not limit the selection of PROG or CQ+PROG for chemoprophylaxis. PMID- 8236465 TI - A prospective study of chloramphenicol induced aplastic anaemia in Nigerians. AB - A five-year prospective study of chloramphenicol induced aplastic anaemia in Nigerians was undertaken. Five proven cases were seen, representing 45.5% of all cases of aplastic anaemia diagnosed during the period of study and 1.0 cases per year. Chloramphenicol induced aplastic anaemia is rare (only 0.002% of all new non-obstetric patients seen in the hospital during the period of study) but preventable. Restriction of the drug or its total withdrawal is recommended. PMID- 8236466 TI - Podoconiosis in the Ethiopian Rift Valley. Role of beryllium and zirconium. AB - A geogenic origin has been proposed in the aetiology of non-filarial elephantiasis of the feet and legs, recently renamed podoconiosis. Soil collected in an area of the Ethiopian Rift Valley, the borough of Ocholo, known for its high prevalence of podoconiosis (5.06%), has been submitted to mineral analysis. High values of sulphur (S), cerium (Ce), lanthanum (La) and neodymium (Nd), typical for basaltic bedrocks, were found. Of special interest were the values for zirconium (Zr) and beryllium (Be), 618 +/- 87 ppm and 4.6 +/- 0.5 ppm respectively, twice as high as those recorded for soils sampled in neighbouring areas where the prevalence of podoconiosis is low. To be noted also, a high content in vanadium, above 250 ppm, in half of the soil samples collected in this region. Year-long exposure of unprotected feet to Zr and Be, known for their ability to induce granuloma formation in the lymphoid tissue of man, and present in a clay rich in colloidal silica particle, highly abrasive to skin, is doubtlessly a factor involved in the development of lymph node sclerosis leading to elephantiasis. PMID- 8236467 TI - The epidemiology of blindness and visual loss in Hamar tribesmen of Ethiopia. The role of gender. AB - We conducted a population-based survey in the Hamar tribe of Ethiopia to assess the problem of blindness and ocular morbidity. Blindness was detected in 1.9% of the Hamar. Among the age group 40 and over < 1.0% of men and 13% of women were blind. Women with low vision or blindness comprised 30% of all women while men with low vision or blindness only comprised 8% of men in the age stratum. Cataract is responsible for the excess blindness in women. Even mild visual disability is less common in men. In Hamar society men have two roles: as herders and as warriors. Warriors with visual loss will be less successful in defending themselves. We suggest that Hamar men who develop visual loss have increased mortality compared to women with visual loss. Vision loss in women does not appear to have life-threatening consequences. Cessation of hostilities between the Hamar and their neighbours could, conceivably, lead to an 'increase' in the incidence of vision loss among men. PMID- 8236468 TI - Attitudes and practices of breast-feeding in Sudanese urban and rural communities. AB - The study was designed to investigate attitudes and practices of breast-feeding in a representative sample of 1039 Sudanese mothers and document corresponding influencing factors in urban and rural communities in six states. Urban mothers were represented by three (high, middle and poor) socioeconomic classes. Almost all mothers (99.9%) initiated breast-feeding, mostly (83.2%) on the first day between 1-5 hours following delivery. Delay of 6 or more hours was practiced mainly by rural mothers (p < 0.001). The breast-feeding rate was 92.0% at 7 months and 65.0% at one year. Four per cent still breast-fed at 2 years. Urbanization affected frequency of breast-feeding adversely; whereas the presence of sore or retracted nipple had a negative effect on its duration (p < 0.02 and < 0.01, respectively). The majority (89.2%) thought that a new pregnancy contraindicated the continuation of breast-feeding and 67.1% reduced or stopped it if the baby had diarrhoea. On the whole, despite the rapid demographic shifts which the Sudan had witnessed recently, positive attitudes to breast-feeding and its duration were still holding. However, there was a tendency towards shorter duration of breast-feeding in urban affluent mothers in comparison to others. PMID- 8236469 TI - Growth of under five-year-old children in Kyeni, Kenya. AB - During a three-months period a cross-sectional study of the measurements of head circumference, mid-upper-arm circumference (MUAC) and weight was performed in 515 under five-year-old children in Kyeni, Kenya. Growth of Kyeni children seems to develop according to international standards for weight-for-age, MUAC-for-age and head circumference and for all parameters the average remains above minus 2 SD of the standard. Despite the presence of anthropometric malnutrition and downward trend in weight-for-age, MUAC-for-age and to a lesser degree the head circumference-for-age, the average growth of the Kyeni children in Kenya is within normal limits of international references. PMID- 8236470 TI - Disseminated strongyloidiasis in a diabetic patient. AB - A case of disseminated strongyloidiasis in an Indian diabetic patient who was not receiving any immunosuppressive therapy, is reported. Strongyloides stercoralis was detected in peritoneal fluid, stools and blood. To our knowledge this represents the first such case to be reported in the English literature. The need for both clinicians and microbiology laboratories to be on the alert for this parasite in patients who are compromised either because of underlying disease or therapy with immunosuppressive drugs, is emphasised. PMID- 8236471 TI - Filarial nephritis: a cause of nephrotic syndrome. AB - A 26-year-old male presented with oedema, massive albuminuria and microscopic haematuria. Kidney biopsy revealed enlarged cellular glomeruli infiltrated by polymorphs and eosinophils with focal fibrin deposits along the basement membrane. Microfilariae were seen in the lumen of few glomerular capillaries. Antistreptolysin titre was negative. The absence of other aetiological factors and presence of microfilariae within glomeruli suggests that nephrotic syndrome may be due to a filarial nephritis. PMID- 8236472 TI - Early repair of frontoethmoidal meningoencephalocele in Bulawayo, Zimbabwe. AB - Frontoethmoidal meningoencephalocele is distinctive in its demographic distribution and its effect on growth of other facial structures. Early diagnosis and referral are of paramount importance. The aim of treatment is to remove the lesion before the deformity disturbs facial growth or alters stereoscopic vision. One-stage, craniofacial surgical intervention is the treatment of choice. PMID- 8236473 TI - Prosthetic valve dysfunction in a Nigerian. AB - A case is presented of a mechanical cardiac valve dysfunction occurring in a 19 year-old Nigerian 7 years after valve surgery and presenting with cardiac failure, supraventricular tachycardia and later, cardiogenic shock but initially masquerading as generalised bleeding probably from Warfarin-drug interaction. The problem of management of an artificial heart valve in our setting is discussed. PMID- 8236474 TI - The retained urinary catheter. AB - Retention of urinary catheters is an increasing complication in our practice. We have reported 18 patients seen over a 6-month period during which 528 catheterizations were done, representing an incidence of 3.4%. Various forms of management were offered and blind percutaneous suprapubic puncture (BPSP) was found to be simple and effective. One out of 13 patients, however, undergoing BPSP developed transient haematuria. The high incidence of malfunctioning catheter balloon mechanisms may be due to poor quality equipment or lack of appropriate storage facilities. PMID- 8236475 TI - Strongyloidiasis in Campinas city region (Brazil). Parasitological and serological data. AB - A retrospective study of laboratory records in a 400-bed university hospital in Campinas city, SP (Southeastern Brazil) suggests that infection by Strongyloides stercoralis is widespread in the region. A prevalence of 10.81% was found in 37,621 stool specimens examined in a two-year period. Parasite-specific IgG antibodies were measured by ELISA in sera from 90 patients with strongyloidiasis. The ELISA detected antibody in 76 (84.44%) patients, indicating that serodiagnostic tests may be helpful in screening patients for strongyloidiasis. PMID- 8236476 TI - Human muscular sarcocystosis in Malaysia. PMID- 8236477 TI - Plasma progesterone and blood metabolite profiles in post-partum small east African zebu cows. AB - Plasma progesterone profiles were used to monitor post-partum reproductive activity in 12 Small East African zebu (Bos indicus) cows allocated to either supplementary or no supplementary feeding (control) with continuous or restricted (twice daily) suckling regimes. Intact bulls were used for breeding. Blood samples were collected 3 times a week for 33 weeks to determine plasma progesterone levels. Weekly blood samples were also used to determine blood metabolite concentrations. Plasma progesterone levels remained below 1 ng/ml in all cows until week 12 post-partum. Only 5 cows showed ovarian activity over the 33 week period. Cows that cycled expressed irregular and short-lived progesterone rises (> 1 ng/ml) lasting 8 to 12 days prior to establishment of normal patterns of progesterone secretion where progesterone levels ranged from 8 to 10 ng/ml in cows with normal cycles. Plasma total protein, albumin, globulin, blood urea nitrogen and glucose levels varied over time without consistent trends, and were not influenced by either supplementary feeding of suckling regimes, nor differed between cyclic and acyclic cows. It was concluded that extended post-partum anoestrus, conception failure and early embryonic mortality were responsible for lowered reproductive efficiency in zebu cows. Blood metabolite concentrations were not good indicators of nutritional status and were not related to post partum ovarian activity. PMID- 8236478 TI - Growth rate and egg production of Creole and Rhode Island red x Creole fowls. AB - In Guyana 3 experiments were conducted using 1,440 chicks and 640 pullets and the same number of hens in a randomised block design to determine the growth rate, egg production, and egg fertility and hatchability of 1/2 Creole x 1/2 Rhode Island Red (CR) and 3/4 Rhode Island Red x 1/4 Creole (CRR) fowls compared with the parental strains of Creole (C) and Rhode Island Red (R) fowls. The growth rate of CR and CRR was higher than C, but lower (P < 0.05) than R. Total egg production per hen housed during the pullet year was 59.5, 106.8, 130.7 and 171.8 for C, CR, CRR and R respectively and they were all significantly (P < 0.05) different. There were no significant (P > 0.05) differences between the 4 strains for fertility and hatchability percentages of the eggs which averaged 87.8 and 88.9 respectively. The level of production was comparatively high and mortality low enough for the crossbreds to be recommended for small holder poultry development, although the feed cost of production would be higher for CRR than for CR. PMID- 8236479 TI - Lactational performance of Jersey cows given Napier fodder (Pennisetum purpureum) with and without protein concentrates in the semi-humid tropics. AB - Two experiments with 12 and 18 lactating Jersey cows respectively were carried out in the coastal semi-humid zone of Kenya to assess the performance arising from the feeding of chopped Napier fodder (Pennisetum purpureum) given ad libitum with and without one of three sources of protein; fishmeal, copra cake and freshly cut Leucaena (Leucaena leucocephala). Each source supplied approximately 300 g crude protein daily. Dry matter intakes of Napier fodder fed alone averaged 7.1 and 5.5 kg in Experiments 1 and 2 respectively. Additional protein did not affect Napier fodder intake, but total intakes of dry matter were higher for the cows receiving the protein supplements, differences which were significant (P < 0.05) in Experiment 2. Average daily milk production from cows fed Napier fodder alone was 6.4 and 4.2 kg in Experiments 1 and 2 respectively. The additional 300 g crude protein increased milk production by 1.0 to 1.6 kg/day, increases which, except that for fishmeal, were significant (P < 0.05). Weight losses of the cows were either reduced or changed to weight gains by the provision of protein. The results are assessed in relation to the energy and protein requirements for milk production. PMID- 8236480 TI - Oestrus ovis myiasis in Libyan sheep and goats. AB - The heads of 1,489 sheep and 320 goats were examined for larvae of Oestrus ovis at 17 abattoirs in northern Libya in July to November 1988. The prevalence of O. ovis in sheep was 22.6% and in goats it was 18.4 per cent. Up to 14 and 11 larvae were collected from individual sheep and goats respectively. All larvae were recovered from the nasal passages and frontal sinuses, but only second and third instars were seen. PMID- 8236481 TI - Tick resistance to acaricides in western Ethiopia. AB - A survey of acaricide resistance was performed on ticks collected from 18 dairy farms and 6 veterinary clinics in western Ethiopia. Boophilus decoloratus collected from most of the dairy farms were resistant to toxaphene, but no resistance was detected in ticks collected from indigenous cattle. There is a need in Ethiopia to replace toxaphene on dairy farms and formulate a rational tick control policy for the whole country. PMID- 8236482 TI - Treatment of theileriosis in crossbred cattle in the Punjab. AB - One hundred and nine cases of bovine tropical theileriosis (Theileria annulata infection) in Punjab State, India, were treated with oxytetracycline (23 cases) or buparvaquone (86 cases). Ages of affected cattle ranged from 6 days to 3 years. Oxytetracycline cured only 7 animals (30.4%), all of them calves below 15 days old, while buparvaquone cured all but one (98.8%), a severely affected 10 day old calf. Cured cattle remained theileriosis-free for 12 to 18 months following recovery. Theileriosis in Punjab is predominantly a disease of young calves that cannot be protected by available cell-culture vaccines. It is suggested that the most economical way to control theileriosis in India would be to immunise calves by infection with sporozoite stabilate and simultaneous treatment with tetracycline, and to reserve buparvaquone for the treatment of clinical cases, in cattle of all ages. PMID- 8236483 TI - Folacin and cyanocobalamin in relation to natural Trypanosoma evansi infection in buffaloes. AB - The haematological values of haemoglobin (Hb), packed cell volume (PCV) and vitamin levels of folacin (folic acid), cyanocobalamin (vitamin B12) of buffaloes, in a surra endemic area of Eastern Haryana, India, were determined. Surra-positive buffaloes had significantly low levels of Hb, PCV, folic acid and vitamin B12. These low levels of folic acid and vitamin B12 may have enhanced the clinical signs of chronic trypanosomiasis, caused by Trypanosoma evansi. PMID- 8236484 TI - Severe complications induced by experimental bacterial superinfection of orf lesions. AB - Twelve goats about 3 months of age were divided into 4 equal groups. Goats in Groups 1 and 2 were infected with orf virus followed by Corynebacterium pyogenes infection of Groups 1 and 3, 3 days after the first appearance of orf lesions. Goats in Group 4 were uninfected controls. Complicated orf lesions which consisted of wet suppurative scabs around the entire lips were observed in goats in Group 1. The lesions persisted for 24 days but were most severe from days 8 to 13. Goats in Group 2 developed lesions typical of orf virus infection that lasted 10 days, while goats in Group 3 developed small nodules of about 1 cm diameter, 48 hours following the introduction of C. pyogenes, which persisted for only 6 days. No lesion was observed in goats in Group 4. Two goats in Group 1 with complicated orf died after 16 and 22 days respectively. PMID- 8236485 TI - Isolation of a rare Salmonella salmonella lohbruegge, and Vero toxin producing Escherichia coli from purging Norwegian dairy goat kids and their crosses. PMID- 8236486 TI - Prevalence of camel brucellosis in Libya. AB - Sera of 967 camels of both sexes were tested for antibodies to Brucella using the Rose Bengal plate test, serum agglutination test and the complement fixation test. The prevalence of positive sera was 4.1 per cent. Samples collected for cultural examination revealed 9 isolates. Five isolates were from milk samples, 3 from aborted foetuses and one from a vaginal swab. All isolates were identified and biotyped as Brucella melitensis biovar 1. PMID- 8236487 TI - Influence of gestation on oestrone sulphate concentration in milk of zebu and crossbred cows and Murrah buffaloes. AB - Single milk samples collected during pregnancy from 128 Sahiwal (zebu), 295 Karan Swiss and 198 Karan Fries cows and 164 Murrah buffaloes were analysed for oestrone sulphate by a sensitive, direct, radioimmunoassay procedure developed in the laboratory. Mean oestrone sulphate levels were below detection limit (< 50 pg/ml) during the first 2 months of pregnancy in all animals. There was an exponential increase in the hormone concentration beginning at the fourth month of pregnancy in cows, although the rate of increase was greatest among Sahiwal cows, followed by Karan Swiss and Karan Fries cows in that order (P < 0.01). An exponential increase in oestrone sulphate levels was also recorded in buffaloes beginning at the fourth month of gestation. However, the mean hormone levels in this species, after initially being lower than Sahiwal and Karan Swiss cows up to 6 months of pregnancy, increased to higher levels thereafter. From this study it was concluded that in addition to genetic factors, environmental adaptation could also influence the oestrone sulphate levels. In addition, the study also provided a basis for pregnancy confirmation by milk oestrone sulphate determination after 110 days in gestation in cattle and buffaloes using a simple, direct, assay procedure. PMID- 8236488 TI - Observations on the incidence of bovine trypanosomosis in cattle dipped in deltamethrin in a tsetse infested area of Zambia. PMID- 8236489 TI - Towards strategic control of ticks in the eastern Cape Province of South Africa. AB - The performance of alphamethrin and flumethrin treated and untreated Bonsmara cows and calves grazing in the thornveld of the Eastern Cape Province was compared over a 2 year period. The economically important tick species occurred seasonally but in insufficient numbers to achieve enzootic stability with respect to babesiosis and anaplasmosis. No differences in live mass were observed for the cows, but the untreated calves were heavier at weaning than the acaricide treated group. Markedly lower numbers of ticks occurred on the calves than on the cows. The implications of the findings are discussed in relation to planning alternative tick control strategies for the region. PMID- 8236490 TI - Antibodies to Cowdria ruminantium in Mozambican goats and cattle detected by immunofluorescence using endothelial cell culture antigen. AB - Endothelial cell cultures, established from bovine umbilical cord arteries and subsequently infected with Cowdria ruminantium, were used as antigen in the indirect fluorescent antibody test. Bovine sera (374) and caprine sera (388) collected in 6 provinces of Mozambique were tested. Overall, 30.4% of goat sera had antibodies to Cowdria, and 43% of sera collected from cattle. North of the River Save, where the tick Amblyomma variegatum is highly prevalent, overall percentages of positive sera were low, 10% in goats and 20% in cattle. However, south of the river where the tick Amblyomma hebraeum is abundant percentages were much higher, 63.5% in goats and 59.4% in cattle. These results are discussed in relation to field observations that clinical disease is rare or absent in the north with enzootic instability in goats and Friesian calves in the south. PMID- 8236491 TI - Observations on the use of Komarov strain of Newcastle disease vaccine in the Sudan. AB - Broilers at 18 days old were vaccinated with a single dose of the K strain of Newcastle disease vaccine administered by different routes. The serological results obtained by haemagglutination inhibition at weekly intervals post vaccination showed that birds vaccinated intranasally (i/n) or intramuscularly (i/m) had antibody titres higher than those vaccinated by the oral method. Furthermore, when challenged 3 weeks after vaccination, those broilers vaccinated i/n or i/m had 75% and 70% resistance respectively, while those vaccinated by the oral or spray methods failed to withstand the challenge. PMID- 8236492 TI - Financial losses caused by ovine fasciolosis in the Ethiopian highlands. AB - An estimate of the economic loss due to ovine fasciolosis in the Ethiopian highlands was made based on available data on mortality, weight loss, reduced reproductive efficiency and liver condemnation at slaughter. The economic effects of fasciolosis were identified and models for estimating the financial loss presented. Ovine fasciolosis losses were estimated at 48.4 million Ethiopian Birr per year of which 46.5, 48.8 and 4.7% were due to mortality, productivity (weight loss and reproductive wastage) and liver condemnation, respectively. These losses can be reduced substantially by fasciolosis control programmes that may include the use of anthelmintics, grazing management and nutritional supplementation. PMID- 8236493 TI - A survey of mineral status of soils, feeds and cattle in the Selale Ethiopian highlands. I. Macro elements. AB - The macro mineral status of cattle in the Selale highlands of Ethiopia was evaluated during the rainy and dry seasons of 1989 and 1990 in terms of mineral concentrations in soils, feeds, blood plasma and faeces on 25 farms. Bone samples from animals of local breeds from a slaughter house were collected during the rainy and dry seasons of 1990. Soils were analysed for pH, organic matter, Ca, P, Mg, Na and K. The results indicated wide variation in the concentrations of minerals on different farms. Available feeds in the area consist of pasture, hay, the straw of barley, oats and teff, and the grains of barley and oats. Pasture grass and other feeds were found to be deficient in Na, P and Mg in relation to dietary requirements. Analyses of blood plasma from crossbred and local cattle showed that a number of samples contained P below the critical level of 1.45 mmol/litre. Effects of year and season were significant for Ca, P, Mg and K. The effect of age was significant for P (P < 0.001). Faecal analyses revealed that about 80% of animals were deficient in Na. Wide variations in the mineral content of soils and lack of significant correlations among soil, pasture and blood plasma indicated that soil and pasture analyses are not reliable in assessing macro mineral status of grazing cattle in this environment. Analyses of bone and blood for Ca and P, blood for Mg and faeces for Na appear to provide better indices. PMID- 8236494 TI - Environmental and genetic effects on preweaning growth performance of hair sheep in Mexico. AB - Data on 4,754 lambs born from Pelibuey (PB) and Barbados Blackbelly (BB) ewes collected over 9 years at the Mococha Experimental Centre in Yucatan, Mexico, were used to determine environmental, dam-breed and sire-breed effects on birth weight (BW), weaning weight (WW) and average daily gain (ADG). Effects of year and season of birth, sex of the lamb, type of birth, breed of dam, breed of sire, their interaction and ewe body weight were included. Heritabilities were estimated for each breed. Season of lambing was a significant source of variation on WW and ADG. Male lambs weighed more than female lambs at birth and weaning. Single born lambs were heavier than twins and triplets. Lambs born from PB dams were heavier and grew faster than those born from BB dams. Heritabilities of BW, WW and ADG for the PB breed were 0.16 +/- 0.04, 0.12 +/- 0.03 and 0.11 +/- 0.03, respectively and 0.04 +/- 0.07, 0.17 +/- 0.09 and 0.15 +/- 0.08 for the BB breed. Heterosis values were close to zero. PMID- 8236495 TI - Factors which affect Gudali and Wakwa calf weights at birth and weaning on the Adamawa Plateau, Cameroon. AB - Birth and weaning data on zebu cattle (Gudali and Wakwa) of north Central Cameroon were analysed. Calf breed, sire and sex had significant effects (P < 0.01 or 0.001) on birth weight while parity and season of birth were not important. Calf breed, sex, season of birth and birth weight had significant effects (P < 0.05 or 0.01 or 0.001) on 8-month weaning weight. Sire and parity had no significant effects (P > 0.05) on weaning weight. Wakwa calves were heavier at birth (24.30 +/- 0.42 versus 22.6 +/- 0.56 kg) and weaning (158.32 +/- 3.46 versus 148.77 +/- 2.88 kg) than Gudali calves. An increase of 1 kg in birth weight led to an increase of 1.27 kg in weaning weight. Calves born at the end of the rainy season were heavier at birth, but were lighter at weaning than those born at the beginning of the rainy season. These results suggest that under extensive low input husbandry conditions calving should be programmed for the end of dry season/early rainy season (breeding during middle to end of the rainy season). PMID- 8236496 TI - Relationship of progesterone in plasma and whole milk of buffaloes during cyclicity and early pregnancy. AB - Blood and milk samples were collected on alternate days from 5 cycling buffaloes on 16 occasions and 2 pregnant buffaloes on 13 and 8 occasions. The blood and milk samples were analysed for progesterone by sensitive direct radioimmunoassay (RIA) procedures developed in this laboratory. The calculated correlation values (r) between plasma and milk progesterone concentrations during cyclicity were 0.8143, 0.5454 and 0.8147 on 0 to 4, 6 to 14 and -6 to -2 days, prior to next oestrus (day 0 = day of oestrus) and the correlations for individual buffaloes during cyclicity were in the range of 0.82 to 0.89 for plasma and milk progesterone concentration. The correlation between milk and plasma levels during early pregnancy in buffaloes was very low (0.1472). The linear regression equation between milk (Y, ng/ml) and plasma (X, ng/ml) progesterone concentration was Y = 1.8 + 3.61X which indicated that the milk progesterone levels were about 4 times higher than those in plasma. It was concluded from this study that in certain situations, such as during cyclicity when large variations occur in corpus luteum activity, the milk and plasma progesterone concentrations from the same buffalo are highly correlated. However, the correlation is very poor when corpus luteum activity remains more or less unchanged, as seen during pregnancy. PMID- 8236497 TI - Cancer mortality in Italy, 1989, and an overview of trends from 1955 to 1989. PMID- 8236498 TI - Hepatoma today: therapeutic experiences of multimodal approach. AB - AIMS AND BACKGROUND: Hepatocellular carcinoma (HCC) ranks third of cancer deaths in China, it kills 100,000 patients every year. In Italy HCC is an increasing malignant tumor and kills about 7000 patients every year. Fortunately, due to a multimodal approach to the treatment of this fatal disease, HCC has been changed from "incurable" to "partly curable". The authors report and comment the methods and the strategies that have been used to increase the cure-rate of HCC. METHODS: The actual approaches, developed in the last thirty years mainly in China, included: screening of cirrhotic patients, early resection, new surgical criteria for cirrhotic liver, early detection of subclinical recurrence and re-resection, multimodality treatment for cytoreduction of huge HCC, sequential resection after cytoreduction of unresectable HCC, targeting therapy using radiolabelled antibodies and lipiodol, transarterial embolization, radiotherapy combined with Chinese herbs, other regional cancer therapy and biological response modifiers. RESULTS: Are that these methods have reached a marked increase series 5-year survival rate and number of 5-year survivors in more expert institutions. CONCLUSIONS: We conclude that further biological and genetic studies on HCC are warranted and that it is mandatory to perform large clinical randomized trials comparing the more promising treatments. PMID- 8236499 TI - Conventional radiographic follow-up in the irradiated lung. AB - AIMS: To highlight the different changes induced in lung tissues by various forms of radiotherapy (RT) according to tumor site and type. METHODS: A retrospective analysis of the roentgenographic evaluation of and long-term follow-up data on 2375 patients who received RT for various intrathoracic and extrathoracic tumors at the National Cancer Institute of Milan. RESULTS: The iconographic patterns of post-RT changes, grouped by site and type of tumor and RT procedure and described in detail, afford deeper insight into a little-known area of lung pathology. CONCLUSIONS: These descriptions of common and uncommon patterns of the irradiated lung as they appear on conventional chest roentgenograms enable the radiologist and radiotherapist to assess exactly the response of tumor and lung tissues and to plan the most appropriate clinical follow-up. PMID- 8236500 TI - Prevention of exposure of young children to parental tobacco smoke: effectiveness of an educational program. AB - AIMS AND BACKGROUND: We conducted a population-based trial to evaluate the efficacy of an intervention aimed at preventing exposure of young children to parental tobacco smoke. METHODS: Of the 1142 eligible couples (parents of newborn babies), 1015 were recruited; the intervention was offered to 402 and not to 613 families. RESULTS: A strong association was found between social class and smoking behavior, in particular smoking during meals at home. The intervention itself had limited effectiveness in decreasing the number of smokers. The effect was stronger in mothers and in higher social groups. Among the "white-collar" families belonging to the intervention group, the proportion of mothers who stopped smoking was 3 times higher than in the control group (not statistically significant). CONCLUSIONS: Educational interventions against smoking should be planned taking into account the difference in efficacy according to social class. PMID- 8236501 TI - Subcutaneous low-dose interleukin-2 plus alpha interferon in advanced malignant melanoma. AB - AIMS AND BACKGROUND: Interferon (IFN) and interleukin-2 (IL-2) have been proven to be active agents in the treatment of malignant melanoma, but the most effective doses of these cytokines were often associated to important side effects and poor patient compliance. Recently, the subcutaneous administration of low-dose IL-2 was found to be a well-tolerated and effective treatment for renal cancer. Since the combination of low doses of IL-2 and IFN has been hypothesized to have synergistic biologic and cytotoxic effects, we evaluated feasibility and patient compliance of a scheme that combined recombinant IFN-alpha (rIFN-alpha) (3 million units by intramuscular injection, 3 times a week) plus low-dose IL-2 (9 million IU, 3 to 5 times a week) administered subcutaneously for 2 weeks every 28 days. RESULTS: Fifteen patients with disseminated malignant melanoma previously treated with chemotherapy entered the study. All but the first 2 self administered the therapy at home and were followed in an out-patient setting. None of them required in-patient care for toxicity. No WHO grade 4 side effects were detected; the only grade 3 side effects were fever and asthenia in 5% of the cycles. Mild hematologic toxicity (grade 2) was observed at the highest weekly dose of IL-2. No major responses were observed in this subset of heavily pretreated patients. CONCLUSIONS: We conclude that the regimen studied is feasible and well tolerated in an out-patient setting, but it is unlikely to be effective. The good patient compliance makes this schedule eligible to evaluate whether IL-2 plus rIFN-alpha can enhance the results of chemotherapy in this disease. PMID- 8236502 TI - M-VECA (methotrexate, vinblastine, epidoxorubicin and carboplatin) in the treatment of advanced urothelial carcinoma. AB - AIMS AND BACKGROUND: Urothelial cancer is a chemosensitive disease. However, cisplatin or anthracycline-containing regimens still provoke severe toxicity mainly due to reduced renal function and poor performance status (PS) of patients. The aim of this study was to verify the possibility of substituting carboplatin for cisplatin and epirubicin for doxorubicin in the M-VAC regimen in order to reduce toxicity and improve patient tolerance. METHODS: Twenty patients with advanced urothelial tract tumors were treated with a chemotherapeutic regimen composed of methotrexate (30 mg/mq iv on days 1, 15, 22), vinblastine (3 mg/m2 iv on days 2, 15, 22), epidoxorubicin (35 mg/m2 iv on day 2) and carboplatin (250 mg/m2 iv on day 2) every four weeks (M-VECA). All patients had bidimensionally measurable disease. RESULTS: Of the 18 evaluable patients, 3 (17%) obtained complete response and 7 (33%) obtained partial response (50% overall response). The median duration of response was 50 weeks (range, 28-88+). Grade III-IV toxicity (leukothrombocytopenia and mucositis) was observed in 20% of cases. Nevertheless, recovery was prompt in all but 2 patients with poor PS who died of nadir sepsis. CONCLUSIONS: M-VECA was an effective regimen for the treatment of patients with metastatic urothelial tumors and was safely employed in patients with a good PS. However, the possibility of substituting carboplatin for cisplatin as neoadjuvant therapy for less advanced stages needs further investigation in randomized studies. PMID- 8236503 TI - Combined use of interferon alpha-2, chlorambucil and prednisone in previously treated patients with low-grade non-Hodgkin's lymphomas. Results of a phase II study. AB - AIMS AND BACKGROUND: To investigate therapeutic activity and safety of alpha interferon (alpha-IFN) in combination with chlorambucil (CLB) and prednisone (PDN), we treated 9 low-grade non-Hodgkin lymphoma patients with clinical evidence of relapsed (5 cases) or resistant (4 cases) disease with such an association. METHODS: In all instances, treatment consisted of alpha-2a IFN administered by subcutaneous route thrice weekly for 3 weeks, CLB, 5 mg/day for 21 days, and PDN, 30 mg three times a week for 3 weeks. Cycles were repeated every 28 days. RESULTS: A well-documented clinical response was observed in 6 (4 CRs+2 PRs) of 9 patients. Interestingly, 3 of 4 CRs were achieved in patients with histologically proven bone marrow involvement. Median duration of response was 18.5 months (range, 4-29 months). Myelosuppression was a common side effect. Two patients experienced grade 3 hematologic toxicity which did not preclude continuation of therapy. CONCLUSIONS: As new purine analogues are not currently available, the combination of alpha-IFN, CLB, and PDN may represent, in our opinion, a valid therapy for patients not eligible for aggressive therapy such as autologous bone marrow transplantation. PMID- 8236504 TI - Radioendocrine therapy of brain tumors with the long acting opioid antagonist naltrexone in association with radiotherapy. AB - AIMS AND BACKGROUND: Malignant gliomas remain untreatable as the different therapeutic combinations are generally only palliative. Recent experimental evidence suggests that endogenous opioid peptides are involved in brain tumor growth. The aim of the present study was to evaluate the effect on survival of concomitant administration of the long-acting opioid antagonist naltrexone (NTX) in patients with malignant astrocytomas treated with radiotherapy (RT). METHODS: 21 patients with high grade malignant gliomas were randomized to receive RT alone or RT plus NTX. The dose of RT was 60 Gy. NTX was given orally at a dose of 100 mg every other day without interruption until disease progression. RESULTS: The objective tumor regression rate in patients treated with RT plus NTX was higher than that of those treated with RT alone but not significantly so. On the contrary, the percentage of survivals at 1 year was significantly higher in patients treated with RT plus NTX than in those treated with RT alone (5/10 vs 1/11, P < 0.05). NTX therapy was substantially well tolerated in most patients. CONCLUSIONS: The finding of longer survival in brain tumor patients treated with RT plus NTX than in those who received RT alone suggests in vivo involvement of endogenous opioid peptides in regulating the growth of malignant astrocytomas. PMID- 8236505 TI - Prevention by L-carnitine of interleukin-2 related cardiac toxicity during cancer immunotherapy. AB - AIMS AND BACKGROUND: Cardiac toxicity has been observed during IL-2 cancer immunotherapy. Because of its trophic action on the myocardial tissue, the use of L-carnitine has been evaluated during IL-2 therapy in advanced cancer patients with clinically important cardiac diseases. METHODS: The study included 30 cancer patients, who were randomized to treatment with IL-2 alone or IL-2 plus L carnitine (1000 mg/day orally). IL-2 was injected subcutaneously at a daily dose of 6 million IU for 5 days/week for 4-6 weeks. RESULTS: The percentage of cardiac complications was significantly lower in patients concomitantly treated with L carnitine than those receiving IL-2 alone (0/15 vs 4/15; P < 0.05), whereas no difference was seen in mean creatine phosphokinase levels on study. CONCLUSIONS: The results would suggest that L-carnitine may be used successfully to prevent cardiac complications during IL-2 immunotherapy in cancer patients with clinically relevant cardiac disorders. Since cardiac metabolism depends mainly on fatty acid oxidation, the stimulatory role of L-carnitine on fatty acid oxidation could explain at least in part its ability to prevent heart disturbances in response to IL-2 administration. PMID- 8236506 TI - Intestinal-type adenocarcinoma of the sinonasal tract: a clinicopathologic study of 18 cases. AB - BACKGROUND: Intestinal-type adenocarcinoma (ITAC) of the nose and paranasal sinuses is a relatively rare tumor. It commonly affects subjects exposed to wood or leather dust. METHODS: The authors present the clinicopathologic findings of 18 cases of sinonasal ITACs and review the proposed histologic classifications. RESULTS: All patients, except one, were males; mean age was 60 years (range, 41 79); in 9 cases an occupational exposure to wood or leather dust was found. Common presenting symptoms were epistaxis, nasal obstruction and rhinorrhea. Histologically, tumors were divided into four groups: well-differentiated (G1) ITACs = 3 cases; moderately differentiated (G2) ITACs = 8 cases; poorly differentiated (G3) ITACs = 2 cases; mucinous (M) ITACs = 5 cases. Immunocytochemically, 16/17 cases were positive for carcinoembryonal antigen, 1/17 for somatostatin, and 0/16 cases for gastrin. CONCLUSIONS: Sinonasal ITACs are aggressive tumors, often diagnosed in a relatively advanced stage. Owing the close similarity of the microscopic aspects, a histologic classification of ITACs analogous to that of colonic adenocarcinomas is proposed. PMID- 8236507 TI - Use of a pool of monoclonal antibodies in diagnosing cells from serous cavities. AB - AIMS AND BACKGROUND: The usefulness of monoclonal antibodies that recognize markers of neoplastic lesions in complementing conventional cytology was evaluated by the avidin-biotin-peroxidase complex, indirect immunoperoxidase technique. METHODS: In order to enhance the sensitivity of the traditional method, a pool of seven combined monoclonal antibodies (Pool C7), which reacts specifically with cells of epithelial origin and is able to distinguish between mesothelial and malignant cells, was tested on cytologic smears of 262 serous effusions. The effusions were benign or neoplastic, mainly from breast, ovary and lung cancers. RESULTS: Immunocytochemical method showed an 100% specificity and 100% of predictivity whereas the sensitivity was 98%, 96% and 95% for breast ovarian and lung carcinomas, respectively. CONCLUSIONS: The results demonstrated that the pool when used together with conventional methods, is useful in analysis of serous effusions in diagnostic investigations. PMID- 8236508 TI - Clinical evaluation of serum aminoterminal propeptide of type III procollagen as tumor marker in gynecologic malignancies. AB - AIMS: The aim of this study was to determine the possible usefulness of the assay of the aminoterminal propeptide of type III procollagen and fibronectin in detecting connective tissue changes associated with gynecologic malignancies. STUDY DESIGN: Serum aminoterminal propeptide of type III procollagen and plasma fibronectin were measured in 36 women with gynecologic malignancies, 20 women with benign gynecologic tumors and 10 healthy women serving as controls. RESULTS: A significant serum propeptide was significantly high in the group with gynecologic malignancies and normal in the benign tumor group. The serum propeptide levels were related to of disease stage and presence of ascites in patients with ovarian carcinoma but not in those with cervical or endometrial carcinoma. In the follow-up study, a favorable clinical response was associated with normalizing propeptide levels whereas in rapidly progressive disease the levels fell initially but rose again. In partial response with ultimate progression, the propeptide concentration decreased but remained clearly above the normal range. No difference in plasma fibronectin was found among the malignant tumor, benign tumor and control groups. CONCLUSIONS: The present study indicates that the aminoterminal propeptide of type III procollagen could serve as an additional, non specific marker to follow the clinical behavior of gynecologic malignancies and consequently of connective tissue metabolism reflecting tumor matrix interaction. PMID- 8236509 TI - Aggressive course of angiofibroma in an HIV-positive patient. AB - We report nasopharyngeal angiofibroma in a 13 year old boy treated with embolization, surgical excision and radiotherapy, which recurred 13 years later. No features of sarcomatous transformation were found, but the tumor had a locally aggressive course, possibly due to the HIV-positivity of the patient who then died of AIDS. The relations between the clinical course of the tumor and severe immunodeficiency are discussed. PMID- 8236510 TI - A calcitonin-producing neuroendocrine tumor of the larynx: a case report. AB - A moderately differentiated neuroendocrine carcinoma of the larynx is described. Although the patient had normal serum calcitonin level 36 months before, its level was elevated at the time the neck node was removed. This study supports the hypothesis that a diagnosis of calcitonin-producing neuro-endocrine tumor of the larynx with increased plasma calcitonin is possible and should be considered to avoid unnecessary thyroidectomy. PMID- 8236511 TI - Tamoxifen-induced immune-mediated platelet destruction. A case report. AB - Drug-induced immunologic thrombocytopenia, a fairly common disorder, is characterized by drug-dependent antiplatelet antibodies that destroy circulating platelets in the presence of the provoking drug or its metabolites. The development of reliable methods for the detection of platelet-bound immunoglobulins causing in vivo platelet destruction, such as the use of monoclonal antibodies tagged with fluorescein and flow cytofluorimetric analysis, has ushered in a new era to differentiate between immune and non-immune thrombocytopenias. A severe thrombocytopenia developed in an elderly female patient treated with tamoxifen, a non-steroidal anti-estrogen drug, after surgery for breast cancer. A tamoxifen-dependent platelet antibody was detected in the patient's serum and linked on the platelet membranes. This antibody reacted only in the presence of the offending drug and showed platelet specificity. Withdrawal of drug restored platelet count to normal levels. PMID- 8236512 TI - Neurophysiological studies of patients with classical phenylketonuria: evaluation of results of IQ scores, EEG and evoked potentials. AB - Neurophysiological studies were conducted in 42 patients with classical phenylketonuria. The results of the intelligence quotient scores, electroencephalogram, visual evoked potentials and brain-stem auditory evoked potentials were evaluated. When compared with the controls, the subjects demonstrated a significant prolongation in VEP P1 and BAEP I-V interpeak latencies and an increase in VEP N1P1 amplitudes. No relationship was found between these pathological responses and metabolic control. However, the observation of normal intelligence quotient scores in 14 out of 18 patients who displayed a pathological prolongation in P1 latencies led us to the conclusion that evoked potentials may have a significant role in the determination of neurophysiological defects and that even cases with good metabolic control may have some obscure neurophysiological dysfunction which should be evaluated more carefully. PMID- 8236513 TI - Deletion analysis of Duchenne muscular dystrophy. AB - DNA of 15 patients with Duchenne muscular dystrophy (DMD) were analyzed for deletions within the DMD gene by using recombinant DNA technology. Deletion frequency was 47 percent and six of the deletions occurred in the region of probe 7 + 8. Only one of the deletions was observed in the region of probe 9-7, and no deletions were found in the region of probe 30-1, 30-2 and 47-4 (5b + 6). The frequency of deletions found in the Turkish DMD patients corresponds to frequencies reported for other populations. PMID- 8236514 TI - Hemolytic-uremic syndrome (HUS): a clinicopathological study of 15 cases. AB - Although hemolytic-uremic syndrome (HUS) is a clinico-pathological entity, renal biopsies are usually not indicated for diagnosis, and therefore, studies concerning the histological aspects of the syndrome are few. This study mainly describes the morphological characteristics of 15 tissue-diagnosed sporadic cases of HUS. The ages of the patients ranged between 10 mos. to 15 yrs., with five being under two. The male/female ratio was 2:3. The prodromal phase was present in 10 patients (67%) with gastrointestinal symptoms in four patients (27%) with neurological symptoms, and in three patients (20%) with upper respiratory infections. Five patients had HUS associated with diarrhea (D+) (three infants and two children), while the remaining ten patients (two infants and eight children) had no diarrhea (D-). E. coli was identified in the stool of four of the D+ cases, one of which was also associated with Shigella. The shortest clinical course was 14 days and the longest 55 days in 13 patients. The disease recurred after three months in one patient, and on three occasions in 15 months after onset of HUS in the other. Fourteen patients died and one biopsy-diagnosed case recovered after the acute phase. All patients had anemia (Hb 3.4-10 g/dl) and acute renal failure. Seven cases demonstrated Burr cells, eight cases had thrombocytopenia and six cases oliguria/anuria. Microscopic hematuria was detected in four cases and gross hematuria in two cases. All patients revealed proteinuria and azotemia (40-200 mg/dl). Five/five (100%) cases had decreased creatinine clearance, 12/14 (86%) cases had increased uric acid levels, 9/14 (64%) cases had an electrolyte imbalance. Light microscopy revealed microangiopathic type involvement of the glomeruli in all cases. According to additional findings, the cases were classed into three histological groups: type 1 showing cortical necrosis (3 cases), type 2 predominant glomerular and arteriolar involvement (11 cases) and type 3 predominant arterial involvement (1 case). All cases were considered primary HUS except for one which was associated with membranous glomerulonephritis. (D+) HUS cases were predominantly of the microangiopathic type, similar to the (D-) group; the latter being contrary to the literature. Hypertension was present in 67% of cases and there was no correlation found between the clinical duration of HUS and the histological type. All five patients studied immunohistologically revealed a nonspecific type fibrinogen deposition. Extra-renal microangiopathy was demonstrated in the adrenals, stomach, pancreas, liver and skin in two necropsies studied. PMID- 8236515 TI - Plasma chromium levels in small-for-gestational-age newborn infants. AB - Plasma chromium (Cr) levels were determined in 24 preterms and 18 full-term newborn infants. There was no statistically significant differences in plasma Cr levels between the preterm and full-term infants. Plasma Cr levels were similar in small-for-gestational-age infants and in infants with hypoglycemia compared with healthy infants. PMID- 8236516 TI - The distribution of haptoglobin types in various regions of Turkey. AB - The distribution of haptoglobin (Hp) types and the gene frequencies of Hp1 in various regions' of Turkey are discussed. The Hp1 gene frequency ranged between 20.4 percent for Turks living in the eastern part of Turkey and 32.6 percent in those who either are at present living in or have migrated from Western Thrace. Generally, the Hp1 gene frequency in the Turkish population is 26.5 percent, a figure similar to that recorded for Asians. Minor variations in the Hp1 gene frequencies in various parts of Turkey may be assumed to have been caused in certain regions by intermingling with the local populations. PMID- 8236517 TI - Protein C and antithrombin III in children with acute leukemia. AB - In this study Protein C (PC) and antithrombin III (AT III) levels in childhood acute leukemia were investigated. The mean PC activity levels in 19 newly diagnosed cases of acute leukemia were significantly lower as compared with the normal controls (p < 0.05). A significant increase was found (p < 0.01) in the patients in remission. Prior to treatment 78.8 percent of patients had decreased PC activity levels, but all patients had normal PC activity during remission. Decreased PC activity levels were found to be independent of the leukocyte count and liver function. No statistically significant difference was found in the AT III antigen levels between the untreated patients, the patients in remission and the control group. Our results indicate that apart from thrombocytopenia, low PC activity levels and alterations in fibrinolysis and coagulation may be responsible for the hemorrhagic manifestations observed in cases of acute leukemia. PMID- 8236518 TI - The effect of exogenous surfactant replacement therapy on lung function. AB - Exogenous surfactant replacement therapy, regardless of type, reduces mortality and morbidity. We have reviewed the existing data to determine the effect this form of treatment has on lung function. Several studies have demonstrated that surfactant replacement therapy improves compliance, but the timing and magnitude of the effect is variable. The effect is dependent firstly on the type of surfactant used, natural surfactant being associated with a more rapid effect than artificial surfactant; secondly whether the surfactant is given as rescue or therapy as prophylaxis and thirdly the gestational age of the infant treated. Preliminary evidence also suggests that surfactant replacement therapy may influence lung function in the long term, since infants treated with surfactant rather than a placebo have lower airways resistance and higher specific conductance at follow-up. These preliminary reports are encouraging because they suggest that exogenous surfactant replacement therapy may have the additional benefit of reducing chronic respiratory morbidity associated with premature birth. PMID- 8236519 TI - Familial sick sinus syndrome in two siblings. AB - Two siblings with sinus node disease are presented. The patients were severely affected and required permanent pacing. A cardiac electrophysiologic study was conducted in Case 2, which revealed an atrioventricular conduction disturbance in addition to sinus node dysfunction. The parents and other siblings showed no evidence of sinus node disorder. The occurrence of this disease in these siblings suggests an autosomal dominant mode of inheritance with variable penetrance. PMID- 8236520 TI - Cardiac arrest: an unusual side-effect of intravenous ornidazole. AB - Nitroimidazoles have been used alone or in combination with other antibacterial agents in the treatment of brain abscess. They are associated with certain adverse reactions related primarily to the gastrointestinal tract and central nervous system. But, as far as we know, cardiac arrest due to ornidazole has not been reported in the literature. In this study, we presented a case in which cardiac arrest occurred as an unusual side-effect of intravenous ornidazole administered to a ten-year-old girl suffering from brain abscess. PMID- 8236521 TI - The role of high-dose intravenous methylprednisolone in the treatment of varicella associated thrombocytopenic purpura. AB - Two children with varicella associated thrombocytopenia are presented. Case 1, evidencing no mucosal or parenchymal bleeding was administered prednisone 2 mg/kg/day therapy after having been put under observation for a period of one week without medical treatment. There was an increase in the platelet count during the period of therapy culminating in a return to normal on the eighth day. Case 2, who was in poor clinical condition at presentation, was treated with high dose intravenous methylprednisolone (HIVMP) 30 mg/kg/day, resulting in the platelet count returning to a normal level (180,000/mm3) on the second day of therapy. HIVMP therapy is recommended in the treatment of life-threatening cases of thrombocytopenia because of its rapid action, limited side-effects, and its low cost. PMID- 8236522 TI - [Status of the antioxidant system and sorbitol pathway of glucose metabolism in diabetes mellitus]. AB - It is found that antioxidative activity (AOA) and activity of glutathione transferase in postmitochondrial placenta fraction of pregnant women with diabetes mellitus is lowered. Depression of AOA in placenta associated with a decrease of UDA and ceruloplasmin amount in blood of the newborns shows the exhaustion of the antioxidative protection system. A decrease of enzyme activity at the sorbitol way of glucose exchange (sorbitol-dehydrogenase and aldose reductase) in placenta reflects accumulation of sorbitol in tissue, which intensifies the damage of membrane structures in placenta. PMID- 8236523 TI - [Structural reorganization and change in the lipid composition of rat liver microsomes in chlorophos poisoning]. AB - The influence of chlorophos on the endoplasmatic reticulum of rat liver has been studied using the methods of gas-liquid chromatography and fluorescence. Experiments have demonstrated an increase of lysophosphatidylethanolamine, total phospholipids, unsaturated fatty acids (C20:4 n6, C18:1 n11, C18:1 n7) and a decrease of phosphatidylserine and phosphatidylethanolamine. Changes in microviscosity and surface charge were also shown. PMID- 8236524 TI - [Preparative isolation of the glycosylated protein and the major core protein of bovine leukemia virus]. AB - A technique for purifying intact bovine leukemia virus (BLV) and a procedure for preparing gag protein (p24) and envelope glycoprotein (gp 51) from intact BLV are described. The preparation method employs solubilization of BLV by n-octyl-beta-D Glycopyranoside and sodium desoxycholate with following ultracentrifugation in a linear 10-60% sucrose gradient. The isolated components preserve their antigenicity. PMID- 8236525 TI - [Rapid turbidimetric micromethod for the simultaneous estimation of parameters of coagulation and fibrinolysis in blood plasma]. AB - A method has been developed to determine six parameters of coagulation and fibrinolysis by means of turbidimetry; 0.1 ml of dissolved plasma after its activation with thrombin and streptokinase is used. Testing conditions which permit performing express diagnostics of haemostasis disturbances and controlling their correction have been optimized. PMID- 8236526 TI - [Intramolecular motion in the protein]. AB - The survey of basic dynamic properties of the globular proteins is presented and the model approach to the study of these properties is considered. Special attention is paid to the function of the intramolecular dynamics in the biochemical processes, in particular, in the enzymatic catalysis. The prospects of the further theoretical and experimental study are discussed. PMID- 8236527 TI - [Glutathione conjugation with diethylmaleate in various brain structures]. AB - It was shown that GSH concentration and glutathione peroxidase activity in hypothalamus (H) and sensomotor cortex (SC) were twice more than in ME. After single or double injection of diethylmaleate (DEM) two-phase change of the GSH level was observed in all the investigated structures. In the first 1-3 h phase, the pool of free GSH decreases by more than 50% as a result of conjugative effect of DEM. In the second phase which is about 3 days long the level of GSH is gradually restored mainly due to the activation of its biosynthesis. In various brain structures conjugation and reduction processes has their own peculiarities: conjugation processes predominate in H and SC while the reduction processes predominate in ME. These processes were almost two times intensified in all the investigated structures under double injection of DEM. PMID- 8236528 TI - [Hydrolysis and the products of 2',5'-oligoadenylate "core" cleavage]. AB - "Core" 2',5'-oligoadenylates resistance to the action of phosphodiesterases was investigated by HPLC and the method of capillary electrophoresis. Oligoadenylate hydrolysis followed by the formation of adenosine and AMP was shown both in the case of incubation with the cells lysate and with the snake venoms phosphodiesterase. Kinetics of products formation in the process of enzymatic hydrolysis was analysed. In contrast to HPLC the method of capillary electrophoresis was observed to be more efficient for 2',5'-oligoadenylates hydrolysis products cleavage. The substance under study was shown not to have sufficient resistance to the enzyme and the latter, probably, hampers its therapeutic use. PMID- 8236529 TI - [Paracatalytic inactivation of pyruvate decarboxylase in the presence of quinones]. AB - Pyruvate promotes the yeast pyruvate decarboxylase inactivation under the influence of substituted p-benzoquinones. Pyruvate decarboxylase activity is not renewed after the removal of low-molecular impurities by gel filtration and subsequent addition of dithiothreitol, thiamine diphosphate, magnesium chloride. The inactivation rate under joint action of 2-methyl-5-isopropyl-p-benzoquinone and pyruvate is regulated by the pseudo-first-order equation. The relationship between pseudo-first-order rate constant and pyruvate concentration takes the shape of hyperbola. The inactivation order with respect to quinone is determined by oxidant concentration and pH value. Maximum pseudo-first-order rate constant values in the presence of the excess substrate and 2-methyl-5-isopropyl-p benzoquinone are observed at pH 5.9-6.0. The data obtained evidence for the fact that during inactivation quinone interacts with "active acetaldehyde" being the intermediate in the process of catalysis with pyruvate decarboxylase. PMID- 8236530 TI - [Mechanism of interaction between quinones and 2alpha-carbanion in the active center of pyruvate decarboxylase]. AB - The kinetics of paracatalytic inactivation of pyruvate decarboxylase by joint action of the substrate and exogenous oxidant p-benzoquinone, methyl-p benzoquinone, 2-methyl-5-isopropyl-p-benzoquinone, trimethyl-p-benzoquinone, tetramethyl-p-benzoquinone has been investigated. Nonlinear correlation between the observed second-order rate constants and redox potentials of quinones has been found. It is supposed that negative deviations from linear dependence was caused by changing the rate determining step by the interaction of quinone with 2 alpha-carbanion in the active centre of pyruvate decarboxylase. According to structure of the oxidant the inactivation rate is limited by one electron transfer or the following protonating of the formed anion radical of quinone. PMID- 8236531 TI - [Kinetic characteristics of enzymes of ethanol metabolism in rats]. AB - Experimental animals were separated as for narcotic sleep length into short sleeping and long-sleeping rat groups and as for alcohol motivation to rats preferring ethanol or water or an intermediate group as well. Alcohol- and aldehyde dehydrogenase reaction maximum speed as well as Michaelis-Menten constants for these enzymes were measured for each of these groups. Alcohol dehydrogenase affinity to cofactor in the reverse reaction was an order of magnitude higher as compared with the affinity to substrate in all the animal groups. Apparent Michaelis-Menten constant values were 1.5-2 times higher in short-sleeping and ethanol preferring rats in the direct reaction. There were no such differences for the reverse reaction. Alcohol dehydrogenase reaction maximum speed for short-sleeping group is close to that for animals preferring ethanol or water. For different forms of aldehyde dehydrogenase an increase of the apparent maximum speeds was distributed as follows: water preferring group < intermediate group < ethanol preferring group. Aldehyde dehydrogenase-1 Michaelis-Menten apparent constants for acetaldehyde were changed in the same way. Thus, data obtained suggest that ethanol metabolism speed along the path alcohol/aldehyde dehydrogenases are close in ethanol-preferring and short-sleeping rats. PMID- 8236532 TI - [Amides of simple and complex ethers of glycolic acid--inhibitors of alcohol dehydrogenase in rat and human liver]. AB - Amides of glycolic ethers and esters have been synthesized and studied as the inhibitors of rat and human liver alcohol dehydrogenase. Amides of glycolic ethers inhibit the rat liver alcohol dehydrogenase activity by 50-85%, amides of esters by 10-40%. An analogous regularity has been established for human liver alcohol dehydrogenase. The inhibiting capacity of the compound for glycolic acid aromatic esters diamides increases depending on the position of two residua of glycolic acid: ortho < meta < para. A decrease in activity of the isoforms of rat liver aldehyde dehydrogenase does not exceed 40%. It is shown that glycolic acid ether amides are noncompetitive inhibitors, amides of aromatic esters in case of the substituent presence in the ortho- and para-positions are competitive or in the meta-position noncompetitive inhibitors. Linear correlation under the comparison of the inhibition constants with the corresponding parameters of hydrophobicity of butyl, 2-methylbutyl and 3-methylbutyl esters of glycolic acid have been found. PMID- 8236533 TI - [Effect of staphylococcal peptidoglycan on the activity of Ca2+-ATP-ase from the sarcoplasmic reticulum in the early stages of ionizing radiation effect]. AB - It is established that peptidoglycan of Staphylococcus aureus possess superficial activity and can render influence on the functional activity of biomembranes. It tends to increase Ca(2+)-ATP-ase activity in the sarcoplasmic reticulum membranes, but no changes have been observed in case of the enzyme solubilization. The effect of peptidoglycan was more expressed, if it was used in the early period of acute radiation injury (1 and 24 h) in a dose of 0.21 Cl/kg for samples with irradiation-induced decrease of Ca(2+)-ATP-ase activity. PMID- 8236534 TI - [Second messenger systems and protein kinase activity in rat thymus and liver in the early stages of radiation injury]. AB - The effect of ionizing radiation (7, 76 Gr) on the content of cyclic nucleotides and Ca2+ ions from rat thymus and liver has been established: it was increased 15, 30 min and 4.6 h and decreased 2, 12 h and 24 h after the irradiation. Changes in activity of cyclic nucleotide-dependent protein kinases and Ca2+, phospholipid-dependent protein kinase correlated with the contents of secondary messengers in the irradiated rats (30 min and 4 h after irradiation) are shown. PMID- 8236535 TI - [Superoxide dismutase in the nuclei of cerebral cortex cells of rats: isolation, properties, effect of ionizing radiation]. AB - Superoxide dismutase (SOD) from rat brain cell nuclei has been distinguished. After chromatography on hydroxylapatite Sephacryl S200 and Mono S SOD can be purified 1200 times. It is Mn-depending enzyme, which has molecular weight 10 kDa and firmly associated with transcriptionally active chromatin. SOD is most active at high ion strength (1.2-2 M NaCl). The more sensitive method for SOD activity determination is suggested (modification of adrenaline autoxidation at pH 10.2). PMID- 8236536 TI - [Role of trace elements in the development of chronic ischemic heart disease]. AB - It was shown that determination of erythrocyte superoxide dismutase activity proves to be the adequate test of antioxidant protection under chronic coronary heart disease as well as the reliable laboratory test for the control of the effect of prophylactic ultra-violet radiation. PMID- 8236537 TI - [Pathways of endogenous malonate formation in the rat liver]. AB - Malonic acid is one of the intermediates of cell metabolism. Malonic dialdehyde and malonyl coenzyme A may be important precursors of malonate. The correlation of biosynthesis intensity and utilization of malonate determines capacity of its intracellular pool. It is supposed that pool capacity is related both to metabolism regulation and to functional condition of organs and tissues. PMID- 8236538 TI - [Influence of ethanol and ethanol-induced lipid peroxidation on the steroidogenic activity of testicles]. AB - Chronic alcohol intoxication results in the diminished testosterone level and activation of lipid peroxidation in rat testes. A significant decrease in 3 beta hydroxysteroid-hydrogenase and 17-dehydrogenase activity has been observed in microsomes. Analogous shifts have been evoked in vitro by the induction of lipid peroxidation in microsomes. That permitted one to suppose that alcohol-induced alterations in tests are partially mediated by induction of lipid peroxidation. PMID- 8236539 TI - [Protein synthesis in tissues of guinea pigs with hypercholesterolemia]. AB - The composition and incorporation of labelled amino acids in electrophoretic fractions of soluble proteins of the liver, brain, heart, kidneys and adrenal tissues of guinea pigs have been studied after 3 months of hypercholesterolemia. Similar quantitative changes in all the investigated tissues have been discovered as manifested by the increase of high-molecular proteins concentration and decrease of low-molecular polypeptides. The decrease of specific radioactivity of total proteins and some electrophoretic fractions has been observed under hypercholesterolemia. Only one of the fractions in the adrenals was characterized by the increase of the label incorporation level. The obtained results indicate that the long-term hypercholesterolemia is accompanied by quantitative redistribution of soluble proteins of the tissues and by suppression of their synthesis. PMID- 8236541 TI - [Hip alloplasty in the 1990's]. PMID- 8236540 TI - [Effect of ionizing radiation on ATPase activity of thymocyte plasma membranes]. AB - Thymocytes were irradiated by the doses of 4-1000 Gr. Activity of Ca(2+)-ATPase and Mg(2+)-ATP-ase, and lipid peroxidation were determined. It is supposed that changes in Ca(2+)-ATPase activity is connected with the adsorption of irradiation energy, while changes in Mg(2+)-ATPase activity are determined by effect of lipid peroxidation. PMID- 8236542 TI - [Unsatisfactory results after repeated revision of hip alloplasties]. AB - Sixty one cemented second revision total hip arthroplasties and 18 cemented third revision total hip arthroplasties were studied with emphasis on cause of failure, complications, risk of further revision, and clinical and radiographic outcome of surviving, not further revised patients. Aseptic loosening was the major reason for both second and third revisions followed by recurrent dislocations. Twenty one second revisions failed again. Postoperative dislocation was the major complication, entered in 10/61 second revisions and in 4/18 third revisions. Ten second revisions were reoperated on without exchange of components, but for reasons related to the implant. Clinical and radiographic outcome was favorable, but must be seen in the light of the high rate of further revisions. In 33/53 second revisions and in 5/13 third revisions left for evaluation, the overall outcome was considered unsatisfactory. Reoperations for failed arthroplasties should be the prerogative of highly experienced centers. PMID- 8236543 TI - [Age and sex specific incidence of primary total hip alloplasty in the county of Southern Jutland]. AB - During the 10-year period 1981-90, 1752 primary total hip arthroplasties (THAs) were performed in the county of south Jutland, Denmark. The annual number of THA increased until a steady state level was reached during 1988-90. The age and sex specific incidences were calculated for this period using the population distribution of the County. The age specific incidences were highest in the age group 70-79 years for both female and males, namely respectively 485 and 410 THAs per 100,000 inhabitants. The overall incidence was 82 THA per 100,000 inhabitants. During the next 30 years, the demand for primary THA in Denmark is expected to increase 32 percent (from 4013 to 5307 THAs) as a consequence of demographic changes. PMID- 8236544 TI - [Induction of obsessions--folie a deux]. AB - The disorder folie a deux is described from the literature with the classical subdivision into four groups, and the characteristic features and involved mechanisms essential to the disorder are presented. Three cases are described, two of them concerning twins, namely a sister-sister and a brother-brother relationship, together with a mother-daughter relationship. The cases show the characteristics in folie a deux: 1. An intimate relationship; 2. Social isolation; 3. Delusions similar in content. Ways of treatment are described. PMID- 8236545 TI - [Nitrosamine formation via non-prescription drugs?]. AB - Both carcinogenic and non-carcinogenic nitrosamines can be formed under physiological conditions in the human body as a reaction between nitrite and a secondary or tertiary amine. A major part of the population is exposed daily through drinking water to high levels of nitrate, which can be reduced to nitrite. Moreover, nitrate and nitrite are both present in vegetables and nitrite is used in food preservation. Dietary exposure to amines is normally well below 100 mg per day, whereas paracetamol and antazoline, both secondary amines, are used therapeutically at much higher doses. Knowledge about the possible interactions between these widely used drugs and the background exposure to nitrite is presently not available. Therefore, an evaluation of the carcinogenic hazard related to this combination is needed. PMID- 8236546 TI - [Focal fatty liver. A dilemma of differential diagnosis in ultrasonography and CT]. AB - Based on four cases and relevant background literature we partly want to draw attention to the occurrence of focal fatty liver (FFL) and its manifold appearances in connection with ultrasonography (US) and computed tomography (CT), and partly to point out possible pitfalls in US-diagnosis of the liver in cases of extensive fatty liver, where the residual foci of normal liver may incorrectly be interpreted as abnormal lesions. US and CT examination of FFL may cause differential diagnostic problems with risk of misinterpretation and erroneous diagnosis, especially if the phenomenon is unknown. In cases of doubt in US examination, where the uncharacteristic findings together with patient information indicate further examinations, additional CT is first of all recommended. If this still brings no clarity to the diagnosis, the clinical condition in every case will prescribe whether to undertake additional CT after a few weeks or immediately carry out US/CT guided liver biopsy. Current as well as formerly presented research shows that FFL is mainly located in the right hepatic lobe, and possible reasons for that are discussed. PMID- 8236547 TI - [Serum sodium as prognostic factor in hepatic coma]. AB - Among 17,628 discharges from the Department of Medical Gastroenterology, Odense University Hospital during the period 1 April 1973 to 31 May 1992, 66 (3.74/1000) discharges in 55 patients were found with the EDP-code 573.00 coma hepaticum. The case records of these patients were examined, and the lowest serum sodium values were compared between the patients who survived hepatic coma and those who did not and were correlated to the total duration of hospitalisation and the serum creatinine value measured at the same time as the lowest serum sodium value. The case records of 54 patients could be found, but in one case there was no serum sodium value; thus, the final material included 53 patients with 53 cases of hepatic coma and 53 lowest serum sodium values, 25 from patients who survived hepatic coma and 28 from patients who died. 15/28 (53.6%) lowest serum sodium values during fatal coma admissions were 119 mmol/l or below, whereas 6/25 values during non-fatal coma admissions were 119 mmol/l or below (p < 0.05). There was a negative correlation between the lowest measured serum sodium value and the total duration of hospitalisation (p < 0.00004). Exponential regression analysis showed negative correlation between serum creatinine and serum sodium (p = 0.0144). CONCLUSION: The correlations mentioned above may raise the question whether serum sodium should be kept at 120 mmol/l or above in patients suffering from hepatic coma. It is not predictable whether such a measure will prevent or attenuate cerebral symptoms and improve survival rate. PMID- 8236548 TI - [Treatment of primary hypercholesterolemia with pravastatin. A placebo-controlled trial]. AB - We present the results of a multicentre, placebo-controlled, randomized double blind comparison of the treatment of primary hypercholesterolaemia with pravastatin (Pravachol), an HMG-CoA-reductase inhibitor. The study was conducted in both general practice and hospital settings. Two hundred and eighty-six patients who despite oral and written dietary instructions and advice on healthy living had elevated total serum cholesterol levels of between six and eight mmol/l were randomized to treatment with either parvastatin (Pravachol) or placebo. The study showed a significant reduction in both total serum cholesterol (of 16%) and serum LDL-cholesterol among those treated with pravastatin (Pravachol), as well as a small increase tin serum HDL-cholesterol of six percent. Advice on diet and healthy living resulted in a reduction of serum cholesterol of only 1.5%. There were no important differences between those treated from the hospital and those treated in general practice. The incidence of side-effects was low and comparable to placebo levels, and only two percent of the pravastatin (Pravachol) treated patients had to have the treatment stopped. All side-effects were reversible. No side-effects relating to the central nervous system were registered, and no elevations of serum creatinine phosphokinase were observed amongst the pravastatin treated patients, presumably because of the drug's selective inhibitory effect on cholesterol synthesis in the liver. PMID- 8236549 TI - [Ambulatory surgery and anesthesia. An inquiry study]. AB - Patients who had undergone surgical procedures as out-patients in either general or local anaesthesia were asked to complete a questionnaire with the aim of evaluating how they had experienced their operation, especially with respect to treatment and prevention of postoperative pain. Over a 12 month period, 851 patients (251 men and 600 women) who had undergone either abdominal, orthopaedic or gynaecological surgery as out-patients were given the questionnaire. Five hundred and fifteen patients (166 men and 349 women), i.e. 61%, answered. Nineteen percent had had their operation performed in local or regional anaesthesia, 30% in a combination of general anaesthesia with local infiltration, the remaining patients were operated under general anaesthesia. Eighty to ninety percent were satisfied with the preoperative information, and 88% were satisfied with the anaesthesia. Sixty-nine percent had had either no pain or almost no pain within the first 24 hours after the operation. Six percent found it difficult to stay home. Sixteen percent needed some kind of medical contact. Ninety-two percent said they would prefer out-patient surgery again as opposed to hospitalization, were they to need another operation. In conclusion, we found that these types of out-patient operations were acceptable to most of our patients (92%). We recommend widespread use of local anaesthetics in combination with general anaesthesia in order to minimize postoperative pain and facilitate the effect of postoperative analgetics. PMID- 8236550 TI - [Ambulatory anesthesia. An inquiry study of patient satisfaction and therapeutic quality]. AB - One hundred and forty-eight patients were anaesthetized in the Day Care Hospital in Ribe and Varde during the period 1st July 1992-30th september 1992. They answered a questionnaire concerning their treatment and their satisfaction. One hundred and twenty-eight patients answered the questionnaire. Sixty-six percent were nervous to some degree, especially the youngest and the women. Nevertheless, 57% men and only 28% of the women wanted sedatives prior to the operation. We concluded that premedication such as midazolam or triazolam should be available. The postoperative pain and side effects were significantly lower using regional analgesia as compared to general anaesthesia. Regional analgesia should be used when possible. There was no detected effect of using naproxen as premedication. The patients were very satisfied with the treatment. If necessary, 96% would prefer to be operated in a Day Care unit again. PMID- 8236551 TI - [Spinal analgesia for cesarean section]. AB - Over a twelve-month period, 47 of a total of 127 women had spinal anaesthesia (SA) for caesarean section (SC). Data from the anaesthesia and the postoperative period were collected and a questionnaire was sent to the mothers after discharge. The analgesia was sufficient in 45 patients. Two had general anaesthesia (GA). One further patient had GA because of overwhelming anxiety. Despite crystalloid preloading, 45% had a significant decrease in systolic blood pressure. Hypotensive cases were treated with a bolus of ephedrine i.v. Forty-one of the new-born babies scored Apgar 10/1 and all scored 10/5. No postdural puncture headaches were observed during the patients' stay in hospital. Forty women answered the questionnaire, and of these, 38 said they would prefer SA in the event of future SC. Six out of eight women who had previously had epidural analgesia preferred SA. We find that SA is a safe, easy, and reliable method for SC, although supplementary analgetic and antiemetic may be needed, but it is mandatory to maintain the systolic blood pressure (cardiac output) close to the preanalgetic values. PMID- 8236552 TI - [Combined spinal-epidural anesthesia for cesarean section]. AB - A retrospective review was undertaken of 61 consecutive cases of combined spinal and epidural block for caesarean section during the period from 27.3.1988 to 13.12.1991. It was found that combined spinal and epidural block was sufficient in 90% of the cases. In 10% it was necessary to give intravenous analgetic supplementation. A high frequency of intermittent hypotension (75%) was found and 15% developed postdural puncture headache. It is concluded that combined spinal and epidural anaesthesia for caesarean section combine the advances of spinal and epidural anaesthesia. However, smaller or Sprotte spinal needles must be used and more vigorous efforts made to avoid hypotension. PMID- 8236553 TI - [Late vascular lesions after cemented hip alloplasty]. AB - We describe a case of a 69 year-old woman with rheumatoid arthritis, who following hip replacement surgery experienced a late migration of the cup of the hip arthroplasty into the pelvis, with ensuing vascular disaster. Procedures for treatment of intra-pelvine cup migration are briefly discussed. PMID- 8236554 TI - [Malignant mesothelioma of the pleura with an unusual course]. AB - An unusual case of malignant mesothelioma of the pleura in a man aged 73 years is described. Four years after debut of symptoms a tumour emerged at the site of a previous pleural drainage and grew during the next two years to the size of a man's head. After removal of the tumour, the histological diagnosis was reviewed and confirmed. Six and a half years after the first diagnosis, the patient is still alive and in a reasonable condition in spite of bilateral pleural involvement. PMID- 8236555 TI - [Perineal hernia after rectal extirpation]. AB - Perineal hernia after abdominoperineal extirpation of the rectum is a rare condition. The most common symptom is perinea/discomfort and bulging. This condition most often develops within a year after extirpation of the rectum. In order to prevent perineal hernia great effort should be taken when closing the peritoneum and the muscular layer towards the pelvic cave. Several repair operations have been proposed in the literature. The authors recommend a transabdominal approach, possibly with interposition of a synthetic mesh. If the perineal skin is damaged, a combined approach with both transabdominal and perineal access should be used. PMID- 8236556 TI - [How much dialysis?]. PMID- 8236557 TI - [Cat-scratch nodes]. PMID- 8236558 TI - [Concerning chronic pelvic pain]. PMID- 8236559 TI - [Pregnancy and eczema]. PMID- 8236560 TI - [Sudden unexpected death in patients with asthma]. PMID- 8236561 TI - [Asthma as immediate cause of death]. AB - In 1987 the Danish National Health Board registered 223 deaths from asthma, and even after correction for errors this is an increase during the last decade. The increase is remarkable, because the antiinflammatory treatment during the same period has been intensified, and a reduction in lethal asthma was expected. Available records of the terminal courses are studied and referred, and it is concluded that well-known risk factors such as severe asthma, previous attacks of acute severe asthma and undertreatment can only explain half of the cases. The onset of maximal asthma treatment is delayed from three to 24 hours, averagely five hours from the first warning sign. The onset of treatment is dependent on the patients' own estimation of their actual state, and it is therefore concluded that better education of asthmatics in objectivizing their lung function is desirable. It is estimated that some asthma deaths might be avoided if the patients are given better and more frequent antiinflammatory treatment, if they use peak-flow meters regularly and if they cooperate with their doctor in working out a personal, specific plan of action in case of warning signs. PMID- 8236562 TI - [Use of glucocorticoids in acute exacerbation of bronchial asthma, parenterally of perorally?]. AB - This is a review concerning the use of glucocorticoids in patients with acute bronchial asthma. Using peak expiratory flow (PEF), forced expired volume in one second (FEV1) and forced vital capacity (FVC) the time course of response shows a significant improvement in one to three hours, maximal effect in eight hours and still significant effect after thirty hours. The smallest dose tested is 15 mg prednisolone eight-hourly orally which seems as effective as methylprednisolone 250 mg six-hourly intravenously. Methylprednisolone-(sodium-acetate) 80 mg given as single dose, intramuscularly seems better than methylprednisolone 32 mg orally twice daily (reducing to 0 mg over eight days) in outpatient treatment of patients not taking glucocorticoids. PMID- 8236563 TI - [Macrosomia. Diagnosis, delivery and complications]. AB - More than 15% of all newborn babies in 1989 in Denmark had a birthweight exceeding 4.000 g, and the incidence is increasing. Risk factors are described in the article. Diagnosing macrosomia before birth is quite difficult, whether using abdominal palpation, ultrasound or measurement of symphysis-fundus height. The foetal weight is often underestimated. Induction of labour due to suspicion of macrosomia is generally not recommended. Macrosomic infants delivered by caesarean section have significantly fewer birth injuries, but the Apgar score is not affected, remaining low. Macrosomia is associated with birth trauma. The severity of the injuries increases with increasing birth weight. More maternal complications are seen as well. Prenatal diagnosis is important. When a macrosomic infant is suspected, ultrasound measurement is recommended after the 38th week. The obstetrician must be trained in the management of shoulder dystocia and should be ready to perform caesarean section. When a birth weight exceeding 5.000 g/5.500 g is suspected, elective caesarean section is recommended. PMID- 8236564 TI - [General practitioners are carrying a heavy load of work, but they are satisfied with their choice of profession]. AB - Seven hundred and seventy-six general practitioners (GPs) sampled randomly from the list of GPs in Denmark were asked about job satisfaction and professional attitudes by questionnaire. The majority (four-fifth) felt that they would decide to become GPs if they were to choose again. On the other hand, two-thirds complained about a workload which had negative effects on their family life. Low job satisfaction was associated with a preference for public employment instead of the current system of private practice. Dissatisfied GPs tended to believe that patients have too great expectations and present problems that they ought to solve themselves. They, also, felt that daily work didn't leave time for preventive medicine and they felt anxiety because of the risk of overlooking serious diseases. PMID- 8236565 TI - [Magnetic resonance spectroscopy of acute cerebral infarctions]. AB - Eight patients with brain infarction were examined serially in the acute phase and one week and two to four weeks after stroke with water-suppressed proton magnetic resonance spectroscopy. Ten healthy volunteers served as controls. The time courses of N-acetylaspartate (NAA), total creatine (Cr), choline containing compounds (Cho), lactate content, and regional cerebral blood flow (rCBF) measured by SPECT were studied. A high lactate level was found in the acute phase. The lactate content decreased to barely detectable levels during the following two to four weeks, while rCBF increased during this period. The content of NAA in infarcted tissue was significantly reduced (p < 0.01) compared to healthy controls, Cr was also reduced (p < 0.02), whereas Cho content did not change. The inverse relationship between lactate level and cerebral blood flow suggests that lactate plays no substantial role in the vasodilatation underlying the hyperaemia that follows reperfusion. The amount of lactate present in the acute phase reflects the severity of ischaemia in the affected region. The content of NAA may be used as a neuronal marker, and thus perhaps as a marker of the effect of future treatment procedures. PMID- 8236566 TI - [Increased incidence of Crohn disease in the county of Copenhagen]. AB - The incidence of Crohn's disease was assessed in a population based study in Copenhagen County from 1962-1987. The incidence increased six fold during the study period from 0.62/10(5) in 1962 to a mean of 4.1/10(5) from 1979-1987, equally in both sexes. The highest incidence was found in the age group 15-24 years: 12.8/10(5) for women and 6.0/10(5) for men (as mean of the period 1979 1987). The prevalence at the end of the study was 54/10(5). The clinical symptoms and extent of disease at diagnosis did not change over the time, but a significantly higher disease activity was found in the 1980's, 74% (1962-1979) vs 84% (1980-1987), p = 0.04. The increase in incidence of Crohn's disease is well established in most parts of the industrialized world. The analysis shows that it is not due to a new disease entity, since the extent of disease, the age and sex distribution, and the symptoms remained unchanged during the 26 year study period. PMID- 8236567 TI - [Lead--a possible risk factor of increased blood pressure and cardiovascular disease]. AB - Many epidemiological and animal experiment studies support the hypothesis of there being a causal association between lead exposure and increased blood pressure/cardiovascular disease. This study includes 1,052 men and women from Copenhagen County, Denmark, who were examined in 1976 and 1981; in 1987, only the men were examined. Blood lead fell by approximately 40% for the men during the 11 year period and by approximately 30% for the women during the first 5-year period. There was a univariate association between systolic blood pressure and blood lead for both sexes in 1976, but it disappeared at the following examinations. The authors found a significant univariate association between changes in blood lead and changes in systolic blood pressure from 1976 to 1987 in the males. All participants taking part in the study in 1976 were followed with respect to hospital admissions and deaths throughout a follow-up period lasting 14 years. There was a significant univariate association between blood lead levels in 1976 and total mortality, coronary heart disease, and cardiovascular disease. However, with regard to coronary heart disease and cardiovascular disease, the association disappeared when controlling for confounders. Blood lead was a significant predictor of total mortality after control for relevant confounders. This study supports the hypothesis of there being a weak causal association between blood lead and blood pressure, total mortality, coronary heart disease, and cardiovascular disease. The importance of this association is very modest for the individual, but the population attributable risk may be considerable. PMID- 8236568 TI - [Early amniocentesis]. AB - Genetic amniocentesis performed at 7-14 weeks of gestation was studied in a series of 138 patients of whom 50 wanted termination of pregnancy (< or = 12 weeks). The material for analysis consisted of 132 samples due to two sampling failures and four samples being handled incorrectly. Forty-eight samples (36 percent) were taken at 7-12 weeks of gestation, mainly transvaginally (36/48:75 percent). The success rate of culture and karyotyping increased with the duration of pregnancy, but was only satisfactory from week 11 onwards. The time until harvest was then 14-15 days. The transvaginal approach is easy to perform and was accepted by the women, but we experienced bacterial or fungal overgrowth in 17 percent of these samples, whereas no infection occurred in the samples taken transabdominally (n = 96). We conclude that genetic amniocentesis is feasible from week 11, but further studies concerning side effects, especially focusing on the procedure-related abortion risk, should be carried out before early amniocentesis is routinely applied. PMID- 8236569 TI - [Pulmonary rheumatoid nodules. A rare disease onset in rheumatoid arthritis]. AB - Rheumatoid nodules are a benign differential diagnosis to neoplasia and infection, when round lesions are accidental findings on chest X-ray. Two cases are reported of rheumatoid lung nodules as presenting features of rheumatoid arthritis. In each case, the diagnosis was verified by open biopsy. PMID- 8236570 TI - [Corneal edema after cataract surgery]. PMID- 8236571 TI - [Official medical diagnosis and interprofessional views]. PMID- 8236572 TI - [Treatment of travelers' diarrhea]. PMID- 8236573 TI - [Use of oral contraceptives and smoking among Danish women]. PMID- 8236574 TI - [Oral contraceptives and cerebral thromboembolism. A Danish case-control study]. AB - A retrospective case control study was carried out in order to assess the risk of developing cerebral thromboembolism by use of low dose oral contraceptives (OC). The subjects were all the 794 women in Denmark aged 15-44 who had suffered a cerebral thromboembolic attack (CTA) during the period 1985-1989, and an age matched randomly selected control population of 1588 women. CTA included occlusion of precerebral arteria (ICD 432), cerebral thrombosis (ICD 433), cerebral embolism (ICD 434), transitory cerebral ischaemia (TCI; ICD 435) and the unspecified group apoplexia cerebri (ICD 436). Of 692/1584 case/control questionnaires sent out, 590/1396 (85.1/88.1%) were returned. Among the cases, 15 refused to participate, 69 had a revised or unreliable diagnosis, 40 had previously had thromboembolic disease, 13 were pregnant, and 152 had a disease predisposing them for CTA, leaving 323 without known predisposition, of whom 320 reported use/non-use of OC. Among 1396 controls, eight either refused to participate, were mentally retarded or resident in foreign countries; 18 returned an uncompleted questionnaire, 17 had previously had thromboembolic disease, 31 were pregnant, and 130 had a disease predisposing them to CTA. Thus, 1198 non predisposed controls were provided, among whom 1197 reported use/non-use of OC. Among 320 cases, 116 (36.3%) were OC users at the time of the CTA, vs. 191 users (16.0%) among 1197 controls, corresponding to a crude odds ratio (OR) of 3.00.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236575 TI - [Oral contraceptives: knowledge and compliance]. AB - One hundred and twenty (120) women, taking contraceptive pills, underwent a structured interview with a view elucidating their knowledge of the physiology of menstruation, the action and side effects of contraceptive pills and their compliance in the taking of contraceptive pills. The most important sources of information were the medical letters in magazines and the women's own doctors, while the teaching in the Folkeskole (primary and lower secondary school) had not had any major influence on the level of information. Well over one third of the interviewed women knew the most important action mechanism of the contraceptive pill, and half of the women could give a satisfactory explanation of the physiology of menstruation. Twenty-four percent (24%) thought that pregnancy could not occur until 1-2 months after the woman had ceased taking the pill. There was high compliance among the women i.e. that their behavior was correct when they had forgotten to take one or two contraceptive pills, when bleeding was irregular, and when beginning on a new package of pills. Eighty-three percent (83%) had experienced side effects that could be related to contraceptive pills. The investigation shows that there is a need for more efficient information about the effects of the Pill and about the physiology of menstruation. PMID- 8236576 TI - [Use of oral contraceptives in Denmark 1980-1990 and smoking habits among fertile women in 1990]. AB - The purpose of this study has been to chart the changes in usage of different types of oral contraceptives (the pill) throughout the 1980's, as well to estimate the extent to which these changes can be expected to influence the incidence of cerebral thromboembolic attacks amongst fertile women. The use of different types of the pill and smoking habits in different age groups are also investigated. The work is based om cross-sectional studies during the period 1980 1990; the use of the pill is documented by national sales statistics from 1980 1990 as well as from questionnaires answered by 1370 healthy women from 1990. The total sale of the pill has been fairly stable throughout the decade, corresponding to a theoretical user percentage of 23-26. The proportion of pills sold containing 50 micrograms oestrogen has fallen steadily from 51% in 1980 to 11% in 1990, while the proportion of pills containing 30-40 micrograms oestrogen (this group includes the mulit-phase pills) has doubled from almost 40% to almost 80% over the same period. Finally, the proportion of minipills has fallen from 10% to 5%. It is estimated that the changed distribution in use of the different types of the pill through the eighties can be expected to reduce the incidence of cerebral thromboembolic attacks among fertile women by 4.5%. Since the 95% confidence limits on the incidence of such attacks among fertile women in Denmark is +/- 14%, one cannot expect to show significant changes in the total incidence of cerebral thrombo-embolic attacks among these women as a consequence of the change in contraceptive pill profile.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236577 TI - [Non-peptide antagonists to angiotensin II receptors. A review]. AB - The renin-angiotensin system is the most important hormone system in the control of blood pressure and electrolyte homeostasis. Pharmacological blockade of the system by means of beta-blockers or ACE-inhibitors is a major tool in the treatment of hypertension and congestive heart failure. Inhibition of the binding of angiotensin to its receptor is, however, theoretically a more direct and selective blocking method. Recently, a series of potent non-peptide antagonists have been developed, which are active when given orally and appear to be promising drug candidates. The clinical and theoretical implications of this discovery are reviewed based upon the present knowledge of the renin-angiotensin system and the available methods for therapeutic intervention in the system. PMID- 8236578 TI - [Loop diuretics. Rational pharmacotherapy]. AB - The pharmacodynamics and -kinetics as well as rational pharmacotherapy of furosemide and bumetanide is reviewed. In renal insufficiency, a reduced response to diuretics is due to altered pharmacokinetics. The optimum dose can be determined within three to four hours by titration and the effect is measured by the amount of excreted sodium. In nephrotic syndrome, both pharmaco-kinetics and- dynamics are altered. The optimum dose is established as above. Starting and ceiling doses are given in tables for both drugs in renal insufficiency and nephrotic syndrome. In congestive heart failure, the difference is greater between oral and intravenous doses than apparent from the bioavailability of the drugs. If potent diuretics are without effect, the heart failure must be treated more vigorously or a combination with thiazides tried out. Potent diuretics are seldom used in the treatment of liver cirrhosis, but, if used, large doses are necessary. Non-steroidal antiinflammatory drugs are usually considered contra indicated in patients with severe renal insufficiency, since the pharmacodynamics of the diuretics are altered. CONCLUSION: The general strategy when using potent diuretics is titration to an effective dose and then using this dose as frequently as needed in order to obtain the desired response. PMID- 8236579 TI - [Short-term and long-term effects on glycosylated hemoglobin after transition from conventional insulin treatment to multiple insulin injection regime in a diabetic outpatient clinic]. AB - In order to examine the short- and long term effects of basal/bolus insulin therapy on the metabolic regulation assessed by glycosylated haemoglobin (HbA1c) the first 201 patients in the diabetes out-patient clinic of the Section of Endocrinology, Department of Medicine, Aalborg Hospital, assigned for this treatment were prospectively studied. In all 201 patients a significant decrease in mean HbA1c value was observed after 21 months on multiple injection therapy (8.5% vs 8.1%; p < 0.001). The higher the HbA1c value on conventional therapy, the greater was the observed decrease in HbA1c following six months on basal/bolus therapy (r = 0.57, p < 0.001). According to an anonymous questionnaire completed by the first 140 patients six months after the start of multiple injection therapy, 97% of the patients preferred to continue with basal/bolus therapy with NovoPen, and 87% felt that they had achieved a better general condition and that the basal/bolus regime made their daily life easier. It is concluded that basal/bolus therapy with NovoPen may combine the attainment of improved metabolic control assessed by a decrease in mean HbA1c in many poorly regulated patients and the subjective experience of an improved quality of life. PMID- 8236580 TI - [Effect of oxytocin on postoperative intestinal atonia after open cholecystectomy]. AB - A double-blind trial was undertaken to assess the effect of oxytocin on the duration of intestinal atony after cholecystectomy. Eleven pairs of patients were either given injections of oxytocin or placebo at regular intervals during the first and second postoperative day. Sequential analysis showed placebo as being more effective, regarding the time until flatus was passed. PMID- 8236581 TI - [Ultrasonically guided drainage of pleural empyema]. AB - Sufficient drainage of the infected pleural space may be difficult without other image guidance than simple frontal and lateral chest radiograms. When the suspected empyema is difficult to locate we recommend insertion of a 10F pigtail catheter guided by ultrasound. This is illustrated by two patient histories. The method is safe, with the advantage of a precise localisation of the drainage catheter in relation to the empyema, the lung and the diaphragm. PMID- 8236582 TI - [Pulse oximetry and obstructive sleep apnea]. AB - We report a case of a 54 years old man, treated with Methylphenidate (Ritalin) because of narcolepsy, who was referred with suspected vertebro-basilar insufficiency. In the hospital, apnoea during sleep attacks were observed. A polysomnography showed marked obstructive sleep apnoea with a disturbed hypnogram, but during the apnoeas the oxygen saturation only decreased by three to six percent. If a patient suspected for obstructive sleep apnoea has only small desaturations, an oximetry can be insensitive as the only "sleep examination", and a full polysomnography is recommended. PMID- 8236583 TI - [For how long should the antidepressive agents be administered?]. PMID- 8236584 TI - [Calcium and vitamin D can prevent fractures in postmenopausal women]. PMID- 8236585 TI - Ultrasonic absorption frequency dependence of two widely used anti-cancer drugs: doxorubicin and daunorubicin. AB - Low intensity ultrasound (approximately 10(-6) W cm-2) in the frequency range 0.5 6.0 MHz was employed to investigate the ultrasound absorption properties of doxorubicin (DOX) at several temperatures. At physiological temperatures, we found enhanced ultrasound absorption from DOX, and its closely related analogue daunorubicin (DNR), in the upper kilohertz frequency range. The findings do not conform to classical theory of ultrasound absorption, thus suggesting an ultrasound coupling with the drug molecules via structural and/or chemical relaxation processes. The absorption spectra are analysed from the point of view of the non-classical theory of sound absorption due to physical and/or chemical relaxations. Only one spectral difference between the two anti-cancer agents is observed, around 2 MHz, and may be attributed to the sole difference in the chemical make-up of the side chain of the two antibiotics. PMID- 8236586 TI - Flow velocity waveforms in the ductus venosus, umbilical vein and inferior vena cava in normal human fetuses at 12-15 weeks of gestation. AB - The objective was to determine the normal Doppler flow velocity waveform patterns in the human fetal ductus venosus, inferior vena cava and umbilical vein correlated with fetal heart rate, and to examine their reproducibility and their inter-relationship at 12 to 15 weeks of gestation. Cross-sectional recordings of 45 normal pregnant women were collected for a data reference range transvaginally and transabdominally depending on fetal size and position. Maximum flow velocity waveforms were obtained from the ductus venosus, the intra-abdominal part of the umbilical vein and inferior vena cava. Time-averaged velocities were calculated in all three vessels together with peak systolic, peak diastolic and time averaged velocities in the ductus venosus and inferior vena cava. Doppler recordings in 21 other patients displayed good reproducibility. Continuous forward flow in the umbilical vein was associated with pulsatile systolic and diastolic forward flow in the ductus venosus. Retrograde flow was present only in the inferior vena cava. Mean time-averaged velocity (SD) in the ductus venosus was 28.8 (6.1) cm/s, in the umbilical vein 9.7 (2.9) cm/s and in the inferior vena cava 10.9 (2.5) cm/s. No correlation could be established between waveform parameters and fetal heart rate. Combined transvaginal and transabdominal Doppler ultrasound allows reproducible blood flow velocity recordings at venous level in early pregnancy. Relatively high velocities were observed in the ductus venosus compared with the umbilical vein and inferior vena cava. Differences in flow velocities in the ductus venosus and inferior vena cava suggest that little or no mixing of blood occurs, a situation well described in sheep. PMID- 8236587 TI - Volumetric arterial flow quantification using echo contrast. An in vitro comparison of three ultrasonic intensity methods: radio frequency, video and Doppler. AB - The two hypotheses presented in this paper are: (1) absolute and relative volumetric flow rates in vessels can be measured by echo contrast time-intensity curves; and (2) echo contrast time-intensity curves generated by different ultrasound backscatter intensity techniques have equivalent capability for flow measurements. A nonpulsatile flow system was built for quantitative ultrasound backscatter measurements from bolus echo contrast injections using two different volumes of mixing. A total of 49 echo contrast bolus injections were made at various flow rates (0.44-2.59 L/min). Ultrasound backscatter time-intensity curves were generated by ultrasound radio frequency, video and Doppler techniques. The rate of backscattered ultrasound intensity washout for each technique (WASHOUT RATE), and relative change in WASHOUT RATE (delta WASHOUT RATE) were compared to the volumetric flow rate (FLOW) and changes in flow rate (delta FLOW), respectively. The relationship between WASHOUT RATE, FLOW and the volume of contrast mixing was studied. A linear relationship was demonstrated between WASHOUT RATE and delta WASHOUT RATE and the corresponding FLOW and delta FLOW by all three methods (r > 0.90 for all comparisons). The WASHOUT RATE was found to depend on the FLOW and the volume of contrast mixing, but the delta WASHOUT RATE was equal to the delta FLOW and independent of the volume of mixing. Time-intensity curves can be generated from different ultrasound backscatter intensity techniques and the WASHOUT RATE correlates well with FLOW. delta FLOW can be determined directly from the corresponding delta WASHOUT RATE. Doppler ultrasound, because of its natural association with the assessment of flow in chambers and vessels, is uniquely suited to assessment of arterial volumetric blood flow in vitro. PMID- 8236588 TI - Quantification of regurgitant flow through bileaflet heart valve prostheses: theoretical and in vitro studies. AB - A theoretical treatment using turbulent jet theory has yielded a new equation for predicting regurgitant flow through bileaflet heart valve prostheses, the most commonly implanted mechanical valve design. Previously reported techniques assuming an axisymmetric jet are not applicable to the slot-like orifices presented in these valves. The equations were therefore rederived in the context of the prosthetic valve geometry. The purpose of this study was to develop such a method and demonstrate its applicability in principle by using in vitro models. The method was validated under both steady and pulsatile flow conditions. Having derived a method geometrically specific to the orifices presented in bileaflet mechanical heart valves, it should be applicable from patient to patient due to the rigid nature of the valve. These idealized in vitro studies, along with the accompanying theoretical derivation, will guide implementation in the clinical setting. PMID- 8236589 TI - Compensation for the signal processing characteristics of ultrasound B-mode scanners in adaptive speckle reduction. AB - A systematic method to compensate for nonlinear amplification of individual ultrasound B-scanners has been investigated in order to optimise performance of an adaptive speckle reduction (ASR) filter for a wide range of clinical ultrasonic imaging equipment. Three potential methods have been investigated: (1) a method involving an appropriate selection of the speckle recognition feature was successful when the scanner signal processing executes simple logarithmic compressions; (2) an inverse transform (decompression) of the B-mode image was effective in correcting for the measured characteristics of image data compression when the algorithm was implemented in full floating point arithmetic; (3) characterising the behaviour of the statistical speckle recognition feature under conditions of speckle noise was found to be the method of choice for implementation of the adaptive speckle reduction algorithm in limited precision integer arithmetic. In this example, the statistical features of variance and mean were investigated. The third method may be implemented on commercially available fast image processing hardware and is also better suited for transfer into dedicated hardware to facilitate real-time adaptive speckle reduction. A systematic method is described for obtaining ASR calibration data from B-mode images of a speckle producing phantom. PMID- 8236590 TI - Non-Rayleigh first-order statistics of ultrasonic backscatter from normal myocardium. AB - Historically, it has been assumed that the first-order envelope statistics of ultrasonic backscatter from myocardium are best described by the Rayleigh probability distribution function (PDF); however, few studies have tested this assumption. This study compares the ability of five PDFs, including the Rayleigh, to describe high-frequency (5-15 MHz) ultrasonic backscatter from in vivo canine myocardium and in vitro human myocardium. The PDFs used have all been previously proposed as descriptions of backscatter from parenchymal tissue or other random media. In this study goodness-of-fit testing of backscatter recorded at several frequencies and insonification angles shows that only in a minority of cases is the envelope. Rayleigh-distributed. In most cases, the K distribution provides the best fit. In addition, computer simulation shows that sparse Poisson scattering can produce K-distributed backscatter. Scattering of this type may contribute to the observed non-Rayleigh character of myocardial backscatter. We conclude that the Rayleigh distribution is not an adequate description of backscatter from myocardium. Non-Rayleigh distributions may offer an additional approach to myocardial tissue characterization. PMID- 8236591 TI - Quantification of ultrasonic anisotropy in normal myocardium with lateral gain compensation of two-dimensional integrated backscatter images. AB - Anisotropy of ultrasonic scattering and attenuation in heart tissue depends on the specific orientation of myofibers with respect to angle of insonification. We used lateral gain compensation (LGC) to correct two-dimensional cardiac images for physiologic anisotropy. Normal hearts excised from three dogs and five pigs were insonified in a water tank with both 2.5 and 5.0 MHz phased-array transducers. Integrated backscatter was measured from a short-axis approach in the anterior wall perpendicular to the principal fiber axis, and in the septum parallel to the fiber axis. The gain in a vertical sector encompassing the septum was adjusted to compensate the image for anisotropy by matching the intensity of scattering from septal and anterior regions. The average gain required to compensate the septum for anisotropy was 16 dB at 2.5 MHz, and 20 dB at 5.0 MHz. Five healthy volunteers underwent imaging with a 2.5 MHz transducer from a parasternal short-axis view. The LGC required in vivo was approximately 16 dB at 2.5 MHz and was equivalent to that required for correction of septal anisotropy in excised hearts. Thus, normal myocardium exhibits substantial ultrasonic anisotropy that can be quantified and compensated for with clinically applicable tissue characterization techniques. PMID- 8236592 TI - Timing of exposures in ultrasonic hemorrhage of murine lung. AB - Pressure thresholds for lung hemorrhage by exposure to low-temporal-average intensity, pulsed ultrasound are of the order of 1 MPa. Earlier evidence suggested that ultrasound modifies the tissue over short periods of time in such a way that the nonthermal action of ultrasound is enhanced. Measurements of thresholds (1) for hemorrhage and (2) for penetration of the hemorrhage through the murine lung in which a given "on-time" was presented to the tissue over periods of time up to 3 min support the hypothesis. PMID- 8236593 TI - Long-term impact of conservative management on localized prostate cancer. A twenty-year experience in Japan. AB - The clinical outcome of 107 patients with localized prostate cancers over the past twenty years was analyzed retrospectively. Immediate endocrine therapy was administered in 55 patients after diagnosis. The other group of 52 patients did not receive any anti-tumor treatment until progression. Overall, 22 patients (21%) died of prostate cancer, while 45 (42%) died of other known causes. During a mean observation period of thirty-seven months, 27 (25%) experienced progression of the disease (local in 9, distant metastasis in 25, and both in 7 patients). The cancer-specific survival rates for these 107 patients were 78 percent at five years and 71 percent at ten years. The timing of endocrine therapy and age at diagnosis did not influence patient's prognosis. Tumor stage failed to demonstrate any prognostic significance after being controlled for other factors including tumor grade. Poorly differentiated histology appeared to be the sole and the strongest predictor for both tumor progression and cancer death. Prostate cancer may not differ significantly among races once it becomes clinically manifest. Expectant management for localized prostate cancer in well and moderately differentiated cancer may be justified because of the higher probability of dying of other intercurrent causes especially in the elderly group of patients. However, definitive forms of therapy should be considered for the group of patients with poorly differentiated tumor who have reasonably long life expectancy. PMID- 8236594 TI - Negative repeat transurethral resection of prostate fails to identify patients with stage A1 prostatic carcinoma at lower risk of progression: a long-term study. AB - Stage A1 (low-grade and low-volume) adenocarcinoma is associated with a low likelihood of progression. Repeat transurethral resection has been used to identify patients at increased risk (residual cancer noted) as well as those at low risk of progression (no residual cancer noted). We recently evaluated the ability of this technique to define a low-risk patient population. We reviewed the records of 24 patients who underwent repeat transurethral resection after they were identified as having Stage A1 prostatic cancer on initial resection (Gleason score < 5, tumor volume comprising < 5% of the resection specimen). Despite no evidence of residual carcinoma on repeat resection, 3 patients (13%) progressed at a mean follow-up of seven years (2 locally, 1 locally and distantly). We conclude that repeat resection does not effectively evaluate the risk of progression and that other techniques including transrectal ultrasonography and serial prostate-specific antigen measurements should be similarly evaluated. PMID- 8236595 TI - Patient compliance in treatment of prostate cancer with luteinizing hormone releasing hormone (LHRH) agonist. AB - Luteinizing hormone-releasing hormone (LHRH) agonist therapy is commonly used as a form of hormonal ablative therapy in patients with advanced prostate cancer. It is important to administer LHRH agonist every four weeks. Any delay of more than two weeks is associated with the risk of disease flare. A retrospective review of two groups of patients were compared. Twenty-five patients treated at the Veterans Administration Medical Center, Louisville, Kentucky, showed that 44 percent missed one or more injections and 24 percent had a delay of more than two weeks after the scheduled time for another injection. Twenty-three patients were treated by a private practice group in Louisville, Kentucky. There were no problems with compliance. An office nurse kept a separate register for patients receiving LHRH agonist therapy and their appointments. For LHRH therapy to be effective, we believe that the level of compliance one could expect from an individual should be determined before instituting LHRH agonist therapy. If good compliance is not assured, alternative forms of hormone ablative therapy may be preferable for patients with advanced prostate cancer. PMID- 8236596 TI - Simple screening for urinary incontinence in the elderly: comparison of simple and multichannel cystometry. AB - Urinary incontinence is a major cause of suffering and disability especially in the elderly. Accurate diagnosis is important to optimize treatment and multichannel cystometry is regarded as the gold standard. Simple cystometry is a technique of bedside cystometry using a single lumen catheter and filling syringe. Simple cystometry was compared with multichannel cystometry in 83 patients (mean age 73.5 years) and was found to have a specificity of 75 percent and a sensitivity of 88 percent for the diagnosis of detrusor instability. Simple cystometry is a quick, easy, and useful test which can be readily performed in the elderly by a doctor or trained nurse. Residual urine measurement, testing for stress incontinence, and estimation of flow rate can be performed at the same time. It can enable the most appropriate treatment to be initiated by improving the diagnostic accuracy of the clinical assessment when more complex resources and expertise is not available. In a proportion of patients further referral and investigation may be necessary. PMID- 8236597 TI - Varicocele: a progressive or static lesion? AB - Varicoceles are associated with ipsilateral testicular atrophy in the adolescent. To determine if varicoceles are capable of causing progressive loss of fertility, we conducted a date-matched, retrospective study. We reviewed 2,989 patients evaluated for infertility at our institution from 1985 to 1990. A total of 285 (8.5%) patients were diagnosed with secondary infertility. A varicocele was identified as the cause of the patient's infertility in 177 (69%) men with secondary infertility. When matched by date to an identical number of men with primary infertility in whom 128 (50%) were infertile on the basis of a varicocele effect, the difference was significant (p < 0.0001). We conclude that a varicocele in some men is a progressive and not a static lesion resulting in the loss of previously established fertility. In addition, varicoceles are not only the leading cause of infertility in men with secondary infertility, but also occur with a greater frequency than in men with primary infertility. PMID- 8236598 TI - Effectiveness of laparoscopic varicocelectomy. AB - Laparoscopic varicocelectomy is a new technique which has been described in a limited number of clinical reports. We reviewed the results of 46 patients undergoing laparoscopic repair of 75 varicoceles over a two-year period to assess the effectiveness of this operative technique. The internal spermatic artery was preserved in 80 percent of the varicoceles and two arteries were present in 5 percent. Our ability to preserve the internal spermatic artery improved significantly with the use of the intraoperative Doppler probe after the first six months of performing this operation (p < 0.01). Nineteen infertile patients had a minimum follow-up of twelve months with seminal improvement in 68 percent and a pregnancy rate of 26 percent. Complications occurred in 2 patients (4%), inferior epigastric vessel bleeding in one, and genitofemoral nerve injury in the other. There was one persistent varicocele (1%). Our results with laparoscopic varicocelectomy are comparable with those reported with standard open surgical approaches. PMID- 8236599 TI - Direct vision internal urethrotomy using topical anesthesia. AB - A consecutive series of 18 patients underwent attempted direct vision internal urethrotomy (DVIU) under topical lidocaine anesthesia. The procedure was completed successfully in 15 of 18 (83%) patients. In these patients 12 of 15 (80%) reported either minimal or no discomfort. Direct vision internal urethrotomy using topical lidocaine anesthesia is a safe and cost effective procedure. PMID- 8236600 TI - Levels of nitric oxide metabolites do not increase during penile erection. AB - Cavernosal smooth muscle relaxation, one of the primary events in penile erection, is initiated by the synthesis and release of nitric oxide (NO) from the neurons of the cavernosa. The present study was undertaken to determine whether or not serum levels of NO metabolites rise during an erection. Since NO is rapidly converted into nitrites and nitrates, we measured these serum levels in the peripheral and cavernosal blood of 15 potent adult male volunteers who were exposed to erotic stimuli in order to elicit a penile erection. Our data demonstrate that both nitrite and nitrate levels in the peripheral and cavernosal blood do not appreciably change during and immediately following an erection. This indicates that the determination of whether or not abnormalities in the synthesis and release of NO play any role in erectile dysfunction will require alternative testing methods. PMID- 8236601 TI - Genitourinary trauma in the pediatric patient. AB - Trauma is the major source of mortality in the pediatric population. A retrospective review was performed on patients admitted to the Children's Hospital and Health Center Trauma Program, San Diego, California, from August 1984 to May 1990. The purpose of this review was to evaluate pediatric trauma and to determine the best treatment and evaluation for genitourinary injuries. Blunt trauma was responsible for 98 percent of the injuries, with renal injuries being the most common. Bladder (7) and male urethral (2) injuries, and vaginal lacerations (8) also occurred. The most severe renal injuries (70%) and all significant bladder and urethral injuries were associated with gross hematuria. Hypotension was present in 31 percent of patients but rarely required surgical exploration for correction. Eighty-six patients underwent radiographic imaging. Computerized tomography (CT) scans demonstrated the most information about intra abdominal solid organ injuries but was inaccurate in detecting bladder or urethral injuries. Genitourinary injury is common in children but rarely requires surgical management. CT scan is the best study to determine extent of solid-organ injury but is inferior to cystourethrography to diagnose bladder or urethral injuries. PMID- 8236602 TI - Nonobstructive dilatation of upper urinary tract may later convert to obstruction. AB - We report on 5 babies found to have pyelocaliectasis or hydroureteronephrosis antenatally who were evaluated soon after birth because of persistent mild or moderate upper urinary tract dilatation. In each instance, vesicoureteral reflux was absent and the technetium-99m diethylenetriaminepentaacetic acid renal scan revealed good function in the ipsilateral kidney. Renogram curves, after furosemide, did not show an obstructed pattern. Indeed, allowing for the dilatation, the drainage patterns were thought to be normal. In each instance the patient represented or was found on follow-up to have increased dilatation. Renograms were then repeated, using the same radiopharmaceutical and dose of diuretic. An obstructive pattern was seen in each instance. These significant observations demonstrate that nonobstructive hydronephrosis, diagnosed prenatally, may later convert to obstruction at the ureteropelvic junction (UPJ) or in the juxtavesical ureter (UVJ). Thus, careful follow-up, for a period yet to be determined, is desirable in patients in whom the initial postnatal evaluation does not demonstrate obstruction by current criteria. PMID- 8236603 TI - Automatic spring-loaded biopsy gun with ultrasonic control for renal transplant biopsy. AB - The automatic spring-loaded biopsy gun with 18-gauge needle was used to perform 20 renal transplant biopsies. A total of 35 needle passes were used during the 20 biopsies to obtain 31 cores of renal tissue (ratio of successful cores to passes 0.88). Nineteen of 20 biopsies (95%) resulted in renal tissue sufficient for diagnosis. One patient experienced gross hematuria that required blood transfusion and resulted in temporary ureteral clot obstruction. We believe the automatic spring-loaded biopsy gun with ultrasonic control allows rapid, accurate, and safe histologic assessment of the renal allograft, and we recommend this system for routine use. PMID- 8236604 TI - Adaptation of Foley catheter for hemostasis during urologic laparoscopy. PMID- 8236605 TI - Fatal perforation of augmentation cystoplasty in an adult. AB - This is a case report of a fatal spontaneous perforation of an augmentation enterocystoplasty in a twenty-nine-year-old T12 paraplegic, thirteen months post augmentation and placement of artificial urinary sphincter. A review of the literature involving 41 episodes in 31 patients is provided with a discussion concerning presentation, etiology, and management. PMID- 8236606 TI - Toxoplasma cystitis associated with acquired immunodeficiency syndrome. AB - Ante-mortem diagnosis of vesical toxoplasmosis in a patient with acquired immunodeficiency syndrome (AIDS), was made by cystoscopic examination and biopsy of the bladder. Vesical biopsies can be helpful in determining appropriate therapy for AIDS patients with urinary tract involvement/disease. PMID- 8236607 TI - Synchronous bilateral testis tumor: mixed germ cell and theca cell tumors. AB - Synchronous bilateral testis tumors of different histologic types are rare. All previous cases have demonstrated germ cell tumors on both sides. The simultaneous appearance of a germ cell tumor and a contralateral non-germ cell tumor has not been reported. We herein report a thirty-four-year-old man who presented with a mixed non-seminomatous germ cell tumor of the left testis and theca cell tumor of the right testis. PMID- 8236608 TI - Lymphoma of urethra in patient with acquired immune deficiency syndrome. AB - Primary involvement of the urethra by lymphoma is rare. We report the first case of a primary urethral B-cell lymphoma as the initial manifestation of the acquired immune deficiency syndrome (AIDS). A fifty-seven-year-old black man was evaluated for obstructive voiding symptoms and bloody urethral discharge. Workup revealed a urethral mass which, on pathologic examination, was a large B-cell lymphoma. PMID- 8236609 TI - Buschke-Lowenstein tumor: therapeutic options including systemic chemotherapy. AB - Verrucous carcinoma of the penis (Buschke-Lowenstein tumor) is a rare variant of squamous cell carcinoma which has been reported to have limited potential for metastatic disease. We report on 2 patients who presented with locally advanced disease after prolonged intervals of neglect. In both cases, the disease was locally aggressive with extensive tissue destruction. In 1 patient, after failure to achieve tumor control with repeated aggressive surgical excision, systemic chemotherapy using bleomycin, cisplatin, methotrexate, and leucovorin led to a complete pathologic response. We believe this is the first reported case in which systemic chemotherapy has been used to successfully treat verrucous carcinoma of the penis. Surgical excision alone was successful in achieving local disease control in the second patient. PMID- 8236610 TI - Laparoscopic nephrectomy for renal neoplasms. AB - Laparoscopic nephrectomy has been shown to be an effective minimally invasive technique for treating benign renal disease requiring surgical excision. However, its application to approach renal malignancy has been limited. Herein, we report on 8 patients with renal tumor who underwent a laparoscopic nephrectomy. All kidneys were removed within Gerota's fascia, and in 3 patients with upper pole tumors, the adrenal gland was also removed en bloc. With limited follow-up of seven to thirty-five months (mean 14 months), there has been no clinical or radiographic evidence of tumor recurrence. We believe that laparoscopic radical nephrectomy with strict adherence to oncologic surgical principles is a practical, less invasive alternative in select patients with renal tumors. PMID- 8236611 TI - Laparoscopic retroperitoneal lymph node dissection in stage A nonseminomatous testis cancer. AB - Open retroperitoneal lymph node dissection in clinical Stage A testis cancer offers the advantages of definitive staging and high cure rates. Observation is elected in various settings to avoid the morbidity and possible sequelae of this procedure. We present a case report of a new procedure, laparoscopic retroperitoneal lymph node dissection, that decreases hospital stay, recovery time, and morbidity while maintaining the advantages of open retroperitoneal lymph node dissection. PMID- 8236612 TI - How to be an effective expert witness. AB - Service as an expert witness can be professionally and financially rewarding. A veterinarian should not assume the responsibilities of being an expert witness, however, unless he or she believes in the case of the party that seeks to hire them. The veterinarian must also be willing to undergo extensive pre-trial preparation to be an effective expert witness. Just as importantly, the veterinarian must possess the temperament and mental discipline to follow the fundamental rules of testifying in order to maintain professionalism and credibility on the witness stand. PMID- 8236613 TI - Veterinary lawsuits. Trends and defense strategies. AB - This article provides a synopsis of malpractice claims presented to veterinarians. An explanation of the coverages provided in a malpractice insurance program is included. The problems facing veterinarians in today's litigious practice environment are outlined. For example, attorneys representing animal owners present veterinary malpractice lawsuits similarly to medical malpractice litigation for the reason that "case law" regarding veterinary malpractice lawsuits is not readily available. PMID- 8236614 TI - Malpractice and liability. AB - Negligence and malpractice are two of the most dreaded words to a veterinarian. This article defines the law of negligence and malpractice. Case results are used to explain some of the problems that have arisen in practice. Ideas and methods that the practitioner can use to avoid lawsuits for malpractice and negligence are included. PMID- 8236615 TI - Aphis and the accredited veterinarian. A partnership in regulatory medicine. AB - APHIS has a long history of partnership with the private practitioner in the veterinary accreditation program. The accreditation program is truly the backbone of the nation's regulatory programs for the prevention, control, and eradication of domestic livestock diseases. The United States' animal export program also relies heavily on the expertise, diligence, and integrity of the accredited practitioner. Accredited veterinarians provide assurance to regulatory officials, and clients, both within the United States and internationally that newly acquired animals will not introduce disease into the livestock population. With the approach of the twenty-first century and the continuing trend toward a global agricultural economy, it becomes even more imperative that APHIS and the accredited veterinarian work together to develop and maintain an active and sound partnership to enhance American agriculture. PMID- 8236616 TI - Wildlife jurisprudence. AB - The area of wildlife law is complex and confusing. Significant international and federal legislation concerning wildlife conservation is reviewed in this article. State regulation, wildlife rehabilitation, and extralabel drug use are briefly discussed. PMID- 8236617 TI - Safety in the veterinary medical workplace environment. Common issues and concerns. AB - This article addresses some of the major areas of concern related to safety in the veterinary workplace. Some practical guidance is offered for setting up programs in areas such as medical waste, hazard communication, general occupational safety and health requirements, shipments of etiologic agents, and pesticides. PMID- 8236618 TI - Zoonoses in practice. AB - Twenty-five zoonoses of importance in the United States are focused upon in this article. Each is presented in outline form. Items are presented in a convenient format that can be of use in explaining zoonoses to owners of animals in which a specific zoonoses has been diagnosed. Also described are specific actions to be taken in diagnosing suspected zoonotic diseases, in preventing transmissions of zoonoses from infected animals to humans in contact or from infected humans to susceptible animals, in notifying physicians of potential human exposure to zoonotic disease, and in reporting to health authorities of those diseases that require notification. PMID- 8236619 TI - The impact of animal welfare and animal anti-cruelty laws on veterinarians. AB - This article discusses the ways in which a veterinarian may be legally involved when there are charges under a humane animal law or a cruelty-to-animals law. Included is a discussion of the elements to be considered when there is prosecution of the veterinarian, action by a veterinary licensing and disciplinary board, action against a client, the veterinarian's duty to warn or disclose, and involvement of the veterinarian as an expert witness. PMID- 8236620 TI - Estate planning for veterinarians. AB - A veterinarian has many tools from which to choose in forming an estate plan. This article defines the tools and explains the major concepts of which veterinarians should be aware in planning for the use and disposal of their assets. Augmented estates, probate and taxable estates, intestacy rules, trusts, and methods of ownership of property are examined as they relate to the practitioner. Included are possible options to consider with a qualified estate planner. PMID- 8236621 TI - Ethics: the why and wherefore of veterinary law. AB - It is impossible to understand one's legal obligations without appreciating the ethical values these obligations are intended to promote. This article presents an overview of the sources of ethical values in veterinary practice. The author explains how legal requirements embody basic moral standards that veterinarians themselves regard as central to their professional role. It is argued that the most effective way of predicting many legal obligations and of motivating oneself to fulfill them is to accept and live by fundamental moral values. The article concludes that the underlying ethical motivation of the law underscores the importance of making attention to ethics an essential part of one's practice of veterinary medicine. PMID- 8236622 TI - Choosing a lawyer. AB - Choosing a lawyer can be confusing. This article discusses criteria to use in selecting a lawyer. Included are appropriate questions to ask during the selection process; considerations related to specialty, expertise, and scope of practice; and determination of fees. PMID- 8236623 TI - Contractual considerations in veterinary practice. AB - Veterinary medicine is a profession based on contract. Many aspects of veterinary practice involve legally enforceable contract obligations. Legal rules established by statutes or court cases govern contract formation, interpretation, and enforcement. This article explains several legal principles governing contract law and applies some of these principles to common contractual settings in veterinary medicine. PMID- 8236624 TI - Employer-employee relations. AB - As an employer, a veterinarian must be vigilant to protect the rights of both job applicants and employees. Federal and state statutes guarantee that applicants and employees will not be subjected to discrimination in any aspect of employment, including recruitment, hiring, promotion, compensation, and termination of employment. An employer may not discriminate on the basis of sex, race, color, national origin, religion, age, or physical disability. In addition, common law tort actions give employees powerful causes of action be against those employers who injure employees. More than ever, employers must be diligent in obeying the law. Employers must also take measures to protect themselves from employee claims by instituting risk management plans and purchasing adequate insurance coverage. PMID- 8236625 TI - Property issues in practice. AB - This article discusses legal issues relating to property ownership. The primary focus is on land use control laws such as zoning ordinances. Cases involving legal questions over the location and operation of veterinary clinics are discussed. An explanation of how land use contracts work and how veterinarians may encounter them is included. The application of nuisance law is reviewed, and representative cases are discussed. Also included is a discussion of property liability issues and what veterinarians can do to avoid litigation. PMID- 8236626 TI - Food and drug regulatory issues. AB - The Federal Food, Drug, and Cosmetic Act does not purport to interfere with the practice of veterinary medicine; however, it does regulate the drugs that veterinarians use in their practice. According to federal law, it is illegal for a veterinarian to use animal drugs in a manner other than that stated on the approved label, to use human-labeled drugs products to treat animals, or to compound drugs for their patients. Because the Act fails to meet the needs of the veterinary profession, the Food And Drug Administration (FDA) has adopted a policy of discretionary enforcement to "allow" veterinarians to use approved animal drugs in an extralabel fashion or to use human drugs to treat animals in certain situations. FDA will not enforce the law against practitioners treating companion, exotic, or sporting animals. Instead, enforcement effort will be directed to treating food-producing animals under conditions most likely to result in illegal drug residues. This is current effort to amend the Act in a way that will make it legal for the FDA to permit extralabel drug use rather than doing so only as a matter of enforcement discretion. Even so, practicing veterinarians will still be limited in how they can use drugs in everyday practice. PMID- 8236627 TI - [Changes in oxytocin and cortisol levels in primiparous cows after changing from 21 days of suckling calves to machine milking]. AB - Four first-calvers of the Black-Pied breed with calves sucking their milk until day 21 of lactation were included in a trial. One suckler cow had four calves. The trial took place within five days which included days 20 and 21 in the first calvers, that means the last two days with sucking calves (designated as day 1 and day 2 of trial-first and second calf sucking) and days 22 and 24, that means the first and third day after the cows were moved to a cowshed (designated as the third and fifth day of trial-first and third evening machine milking). The first calvers were separated from the calves and moved to a cowshed at 8 o'clock a.m. on day 3 of trial (Tab. I). The responses to calf sucking were investigated after four-hour prevention of calves to approach the cows (12.00-16.00 p.m.). Blood was sampled with a catheter which was introduced into the vena jugularis a day before the trial outset. Blood samples for cortisol determination were taken in two-hour intervals (when the cows were moved to the cowshed twice in half-an-hour interval and once in an hour interval). Blood samplings were effectuated by day, before, during and after milking, or sucking. Machine milking was finished within 7 to 8 minutes. Two to four calves were intensively sucking a cow within the first 15 minutes, then sucking was irregular. Plasma samples were deep-frozen and stored at -20 degrees C until determinations were performed. Oxytocin and cortisol in the blood plasma were determined by radioassays. In the first evening machine milking (day 3) the second cow yielded 8.23 kg milk while on day 5 in the third milking it was 7.98 kg. The other first-calvers gave by 30-45% less milk on day 3 in comparison with evening milking on day 5 (Tab. II). A trend of the higher average values of oxytocin was observed in machine milking on day 5 of trial if compared with the values recorded during calf sucking, or the first milking (day 3), Fig. 1a, Tab. III. The mean oxytocin concentration during 7-minute milking (12.39 +/- 5.81 pg/ml) was significantly lower in the first-calvers on day 3 in comparison with the mean values recorded on day 5 (28.01 +/- 18.56 pg/ml; P < 0.001), or during sucking (18.32 +/- 7.15 pg/ml; P < 0.01).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8236628 TI - [Paroxysmal hemoglobinuria in calves and its effect on hematologic and acid-base profiles]. AB - Water intoxication is considered to be one of the possible causes of haemoglobinuria in calves. As to predisposing factors, prognosis and particularly differential diagnosis, uncertainty is rather frequent. Therefore we tried to provoke this state in an experiment, in which the disiony-induced changes of the acid-base balance (pH, pCO2, HCO3, pO2, BE, SAT of venous blood) were determined in addition to clinical observations (general behaviour, respiratory and pulse rates, body temperatures) and blood analysis (red and white blood cells, PCV and haemoglobin). Experimental paroxysmal haemoglobinuria was induced in eight 2 months-old male calves of the Black Pied breed weighing 55-70 kg that were held on a green diet. After the animals had been given cold water (12-14 degrees C) at an amount of 12% of their body weight, increased volume of the abdominal cavity, muscle tremor, and a stooping posture could be seen, and in 45-60 min. following water administration the first spontaneous haemoglobinuria occurred. Except the first 20 minutes following water gavage, when pronounced tachycardia accompanied by arythmia (100-130 per min.) was recorded, a tendency towards hypothermy, mild bradycardia and bradypnoe was observed; correction of these values occurred within 24 hours. Haematological indices revealed a hydraemic trend that was most pronounced and long-lasting in haemoglobin. Twenty-four hours after water gavage haemoglobin, red blood cells and PCV values were still below their starting levels; the tendency toward leucopoenia changed into leucocytosis. The acid-base balance revealed a slight acidemic trend with decreasing pH, actual bicarbonate and BE levels in the first hour; later, equilibration and increase within the physiological range occurred.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236629 TI - [Experimental paroxysmal hemoglobinuria in calves and selected biochemical indicators in the blood and urine]. AB - When examining diseased calves, sporadically pronounced haemoglobinuria with dark red urine can be observed. In serious cases the clinical picture may be manifold but peculiar; in easy cases, however, when there are no distinct clinical symptoms, a larger scale of examinations is needed to aid differential diagnosis. Eight roughage-fed bulls aged two months, weighing 55-71 kg were used in this experiment. Selected biochemical indices of the mineral, enzymatic, hepatic, energetic and urinary profile were determined in the blood serum and urine of the animals. After the administration of cold water at an amount representing 12% of the animal's body weight, ionogram values were determined. In all indices a positive correlation with hydraemia and a decrease in Na, Cl, Ca, Mg and P levels were observed. Correction of the above levels occurred within 24 hours, with the exception of Na and P concentrations that did not reach starting values. As to the enzymatic profile (AST, ALT, GGT), no pronounced disturbances could be observed. The most profound changes were seen in AST activity that increased in the 5th hour of the experiment. A slight tendency towards hypoproteinaemia was observed to continue even in 24 hours. Hypoglobulinaemia reached its starting value in the 24th hour while simultaneously albumin levels slightly increased. The increasing bilirubin levels reached their maximum in the 5th and 6th hour; correction of the former occurred within 24 hours. The urinary profile revealed polyuria, aciduria, aquaeous urine and haemoglobinuria, the latter reaching its peak between hours 1 and 3 following water administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236630 TI - [The effect of primary BHV-1 infection on the dynamics of T and B lymphocytes in the peripheral blood and levels of specific serum antibodies in calves treated with glucan]. AB - In the present study we investigated the dynamics of circulating T and B lymphocytes and serum specific antibodies in calves experimentally infected with IBR virus (I group) and in calves administered glucan (seven days before infection) and the infected (GI group). The percentages of T and B lymphocytes in the peripheral blood were determined from analyses by rosette methods; the titer of serum anti-IBR antibodies was determined by virus-neutralizing test on cell cultures. The dynamics of the percentage of circulating T lymphocytes showed a similar decreasing trend in both groups, with significant values on days 3 to 5 after infection (AI). In comparison with the calves of I group, the outset of T cell reduction was found to be less pronounced (within the first two days after infection), with a statistically significant difference on day 2 AI (P < 0.05), Fig. 1. A decrease in the percentage of T lymphocytes was related to an increase in the percentage of circulating B lymphocytes, with maximum on days 3 and 4 AI (P < 0.05), Fig. 1. All the calves before experiment beginning were free of serum anti-BHV 1 antibodies. They started responding to the experimental infection with IBR virus by production of serum antibodies between week 1 and 2 AI. The dynamics of serum anti-IBR antibodies showed an identical course in both experimental groups, with the more pronounced outset (P > 0.05) of immunological response in the calves of GI group (Fig. 2).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236631 TI - [The ovarian response and conception rate in cows during experimental nitrate loading]. AB - The effects of potassium nitrate were investigated on ovarian activity, especially on ovarian activity, especially on follicle ripening and ovulation, conception rate and progesterone levels in the milk of cows with cloprostenol induced oestrus (Oestrophan Spofa) at a rate of 500 micrograms i.m. Potassium nitrate was administered at an amount of 150 g per head/day, and this amount increased by 50 g every other fortnight to the total rate of 300 g per head/day. In a group of 11 cows exposed to potassium nitrate load, clinical examination of ovaries on day 8 after oestrus revealed the presence of CL in seven cows (63.6%), persisting follicles in two cows, and two cows did not have any CL or follicles (Tab. I). After the first insemination three cows got in calf (27.3%), and after all inseminations it was 10 cows in total (90.9%) on average in 24.4 days after cloprostenol administration, with SP 96.7 days and insemination index 1.7. No ovulation disorders were found in the control group. All five cows got in calf after the first insemination in three days after cloprostenol administration with SP 61.5 days and insemination index 1.0 (Tab. II). The progesterone levels in milk were 22.3 and 21.1 ng/ml before the second administration of cloprostenol, 2.7 and 2.1 ng/ml in oestrus and at the time of A.I., and 16.6 in eight days after insemination in cows exposed to nitrate load, and finally 21.7 ng/ml in the control group (Tab. III). Ovulation disorders were easily detectable by a clinical check of ovaries on day 8 after oestrus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236632 TI - [Biochemical indicators of nitrogen and energy metabolism in the plasma and blood of beef bulls during the grazing season]. AB - Concentrations of total proteins, urea, total lipids in the blood plasma and glucose concentrations in the blood of beef bullocks were followed after their transition from winter feed rations to pasture, and in the subsequent grazing cycles within two years. Blood was taken by punctures from the vena jugularis from the same six bullocks all the time. In the 1st year of observation, blood sampling took place a week before the bullocks were moved to pasture, in the 2nd year of observation blood was taken twice in weekly intervals before pasture. During grazing, biological material was sampled in weekly intervals between 1st and 5th week (May--cycle I, June--cycle II), then in the tenth week (July--cycle III) and in the 14 week (August--pasture cycle IV). The concentrations of total proteins in the blood plasma of beef bullocks (Fig. 1) insignificantly increased after transition from indoor housing to pasture, this increase continued in the second year of observation for two weeks after transition to pasture, and in the first year of observation for four weeks with a break in the 2nd week. In the subsequent period, the concentrations of total proteins decreased to the values approaching the starting values. The concentrations of plasma urea in beef bullocks (Fig. 2) significantly increased in both years of observation after transition from stable fattening to pasture (p < 0.01 and p < 0.05, resp.). In the second week after transition to pasture, further highly significant increase in the concentrations of plasma urea was observed in both years of observation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236633 TI - [Arachidonic acid in the liver, ovary and uterus of cows with various reproductive capability]. AB - Research was carried out at two localities ecologically different, i.e. in the "relatively" clean region of Klatovy, designed according to the standards of the Czech Ministry of Environment as a region with above-average quality environment and in the considerably affected region of Prague-East, designed as a region with below-average quality environment. The first group included 33 cows from herds with good fertility (average conception rate after 1st insemination 61.5 and service period 88 days) and the dairy cows were eliminated for reasons different as reproductive disorders (Klatovy), in the second group all 18 cows were eliminated because of reproductive disorders (Prague-East) and they were from the herds with much lower fertility (average conception rate after 1st insemination 46.9% and service period 107 days). Samples of liver, uterus and ovarian tissues of the cows were processed according to the method after Jary et al. (1987), analysed on the gas chromatograph Varian--Aerogramm 2100. The data obtained were processed on a computer IBM PC--AT with a numeric co-processor, and the programme Quatro Pro, version 2.0 was used. Differences in average amounts measured in both regions are objectively characterized in Figs. 1, 2 (with marked scatterings) and correspond to the amount given in Tab. I. As regards the saturated acid, stearic acid and oleic acid, no remarkable differences were found according to the localities (Klatovy and Prague), with the exception of the average contents of those acids in uterine horns. In that case the region of Klatovy is dominating.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236634 TI - [Manganese levels in biological materials from dairy cows]. AB - Manganese in serum, hair and excrements of 2,720 dairy cows was determined under the practical conditions of animal husbandry. Mn content in the above-mentioned materials indicated the state of sufficient supply in 1,480 dairy cows and Mn deficiency in 1,240 dairy cows. The results of Mn content in the followed materials obtained from dairy cows in a different state of organism supplying are presented in Tabs. I-III. Sufficiently supplied dairy cows (Tab. I, Ist group) displayed a significantly higher (P < 0.01) content of Mn in serum equal to 0.042 mg/l with 95% interval of reliability, of the mean value (95% IR) 0.039-0.046 mg/l compared with the pooled group of deficient dairy cows (K) with the content of Mn in serum 0.037 mg/l and 95% IR in the range of 0.034-0.040 mg/l of serum. Mn content in hair (Tab. II) of sufficiently supplied dairy cows (Ist group) exceeds the limit value 7.0 mg/kg DM as for both the mean value and the 95% IR. The content of this element in dairy cows in the state of primary and secondary deficiency (groups II and IV) lies below the limit value within the range. The IIIrd group of dairy cows, which was in the state of depletion, contained Mn above the limit value. The dynamic process of deficiency development is in progress in this group of dairy cows, indicated by a reduced content in serum and excrements and for time being a still sufficient content in hair which is expected to decline with time and further growth and eventually lead to primary deficiency.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236635 TI - [Chromosome aberrations in the peripheral lymphocytes in breeding sheep stock]. AB - The health of animals depends not only on the level of hygiene on the farms but also on the ability of the individual to withstand diseases and to face various unfavourable effects of the environment. On the cellular level, chromosome analysis in the peripheral lymphocytes of farm animals is used to determine the effects of various environmental factors. According to present knowledge the frequency of chromosome aberrations is considered to be a quantitative index of genetic damage. The present work reports on in-vitro observations of structural aberrations in the peripheral lymphocytes of Merino and crossbred sheep kept under good hygienic conditions. Comparison of the incidence of aberrant cells (AB.C.) was carried out in two seasons of the year (Tab. I). There was not a significant difference (P > 0.05) between the spring (0.96%-AB.C.) and autumn findings (1.59% AB.C.). In these seasons, chromatid breaks (B1) were the most frequently recorded structural aberrations. Comparison of the three age groups (comprising animals aged 1-2, 3-4 or 5-9 years) did not reveal any significant differences in the AB.C. percentage (P > 0.05, Tab. II). PMID- 8236636 TI - [The effect of experimental aerobic stabilization of swine slurry on survival of Salmonella typhimurium and Ascaris suum]. AB - Laboratory aerobic mesophilic stabilisation (fermentation) of pig slurry reduced the survival time of S. typhimurium, compared with their prevalence in anaerobic excrements. The decimation time T90 for S. typhimurium, was 32 hours. The effect of aerobic stabilisation on the survival of nonembryoed eggs of the model helminth A. suum was lower than that observed in Salmonella. After 54 hours of aerobic stabilization of slurry only 32% of eggs were devitalized. Stabilized control group did not develop 13% of eggs into their embryonated stage. PMID- 8236637 TI - [Serologic screening for the occurrence of Leptospira antibodies in small wild mammals]. AB - During the five years (1986-1990) free living small mammals were examined serologically for the presence of antibodies to leptospires. Standardized techniques were used in the serological examinations (Sebek, 1979). A total 4634 specimens representing 15 mammalian species were examined. During the studied period, higher occurrence of small terrestrial mammals was noticed in the years 1987-1988. In these years, 61% of specimens out of the total number of trapped animals were examined. The field mice (Apodemus sylvaticus and Apodemus flavicollis) and the common vole (Microtus arvalis) were most frequently examined species, their proportion in the total number being 35.4% and 28.4% respectively. Antibodies to leptospires were demonstrated in 557 cases, i.e. in 12.06%, in specimens representing 10 mammalian species, the higher percentage was recorded only in the year 1988, when antibodies were detected in 15.4%. In 550 cases, i.e. 98.7%, the antibodies were to leptospires of the serovar grippotyphosa and only in 7 cases, i.e. 1.3% to leptospires of the serological group Sejroe. Antibodies to leptospires of the serovar grippotyphosa were found in specimens captured in all localities studied. Antibodies were most frequently detected in the specimens of the common vole (Microtus arvalis) in 6.1% and in both species of field mice (Apodemus sylvaticus and Apodemus flavicollis) in 3.4% of the total number of animals examined. Antibodies to leptospires of the serological group Sejroe were found only in blood sera of the house mouse (Mus musculus) and only in one locality studied. Our results confirmed some previously known facts concerning the distribution and structure of leptospirosis foci on this territory. The study of population dynamics should not be neglected because it is directly proportional to the infestation of individual species as we have evidenced. This fact is most expressive in main reservoir species. PMID- 8236638 TI - [Clinical experience and comparison of ketamine-medetomidine with ketamine xylazine anesthesia in the African wild dog (Lycaon pictus) in captivity]. AB - The effects of two mixtures, ketamine-xylazine and ketamine-medetomidine, were compared in anesthesia of African wild dog (Lycaon pictus) in the Zoo park at Dvur Kralove; the effects of these combinations were also investigated on the triad values and on the basic hematological (red blood counts, hematocrit, hemoglobin content, derived parameters MCM, MCHC, MCV, white blood counts, differential blood counting) and biochemical (total proteins, glucose, creatinine, urea, cholesterol, magnesium, calcium, phosphorus, chlorides, sodium, potassium, alkaline phosphatase, gamma-glutamyl transferase, alanine transferase, aspartate transaminase) parameters of blood and blood serum in anesthesia. Tab. I shows the reasons for anesthesia in 68 individuals of African wild dog in the years 1980-1990. As for both mixtures, application of drugs with a blowpipe was used. In the anesthetized animals, the outset of ataxia, laying down and the outset of sleep were followed (Tab. II), as well as the time of wakening up without and with antidote administration (Tabs. IV and V). The outset of ataxia was fast 1.5 +/- 0.6 min) in the ketamine-xylazine mixture administered at doses of 5.07 +/- 1.16 mg/kg ketamine and 2.11 +/- 0.53 mg/kg xylazine, similarly like lying down (3.2 +/- 1.0 min) and loosing sensation (6.3 +/- 1.6 min). At the start of the drug action, vomiting was often observed, and sometimes in the first ten minutes after drug harpooning short clonic convulsions of the limbs or the whole body occurred. In further course, immobilization and anesthesia were complete and satisfactory in all cases. The first reactions to outer stimuli during wakening up without antidote administration were observed in 135 +/- 11.9 minutes while the animals stood up in 210 +/- 44.5 minutes after drug harpooning (Tab. IV). When the nonspecific antidote xylazine-yohimbine was used, the first reactions after i.m. instillation at a dose of 0.31 +/- 0.02 mg/kg appeared in 20.3 +/- 0.57 min, after i.v. instillation at a dose of 0.11 +/- 0.17 it was in 9.3 +/- 4.16 min after antidote administration. The animals stood up and started walking in 53.3 +/- 24.3 min after i.v. instillation (Tab. V). No significant changes (Tab. III) were observed in the values of temperature, breathing rate and pulse rate when the variations of triad values were investigated in five individuals within the first 30 minutes (up to 10 min, 20 and 30 min) after drug harpooning.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8236639 TI - 16 kDa envelope proteins in non-Serpulina hyodysenteriae spirochaetes isolated from pigs. AB - Spirochaetes isolated from field samples of diarrhoea, 'colitis' and mucoid diarrhoea from pigs were examined by a series of cultural, biochemical and serological tests. In addition sodium dodecyl sulphate polyacrylamide gel electrophoresis was used to determine whether the organisms possessed a 16 kDa protein thought to distinguish Serpulina hyodysenteriae from S innocens. Spirochaetal isolates which differed culturally and biochemically from S hyodysenteriae were found to possess a 16 kDa protein. One of these isolates was examined by electron microscopy and found to have an ultrastructure differing from that of S hyodysenteriae. Antiserum to the 16 kDa antigen of S hyodysenteriae reacted with isolate S80/5, the homologous strain, and with B78, the type species, but not with the 16 kDa antigens of the field isolates considered to be S hyodysenteriae or with the non-S hyodysenteriae spirochaetes. It was concluded that there may be a family of 16 kDa proteins located on the envelope of various spirochaetes responsible for diarrhoea in pigs. PMID- 8236640 TI - Comparison of the postoperative analgesic and sedative effects of carprofen and papaveretum in the dog. AB - Forty dogs undergoing a variety of surgical procedures were assigned randomly to one of two groups. All the animals were premedicated with acepromazine (0.05 mg/kg bodyweight) intramuscularly, and anaesthesia was induced with thiopentone sodium, or propofol in the case of lean animals, and maintained with halothane in an oxygen/nitrous oxide mixture using a non-rebreathing circuit. The dogs in group 1 were given papaveretum (0.2 mg/kg) slowly intravenously within 35 minutes of induction of anaesthesia and the dogs in group 2 were given carprofen (4 mg/kg) in the same way. The dogs were scored for sedation and pain by a trained theatre nurse, who did not know which group they belonged to, using a visual analogue scale, at 15, 30, 60, 120, 240 and 360 minutes after the halothane was switched off at the end of the procedure. Nine of the dogs were withdrawn from the trial (eight of them from the papaveretum group) because of inadequate pain relief and these animals were given pethidine (3 mg/kg intramuscularly) which produced adequate analgesia within 15 minutes in all but one case. Carprofen provided profound analgesia which was as effective and of longer duration than that produced by papaveretum, and was associated with significantly less postoperative sedation and a quicker return to the normal conscious state. PMID- 8236641 TI - Mycobacterium kansasii infection in a llama. PMID- 8236642 TI - Corpora amylacea in seminal vesicles and ampullae of tsessebi antelopes (Damaliscus lunatus). PMID- 8236643 TI - Low prevalence of feline viral infections in northern Greece. PMID- 8236644 TI - Regurgitation in a sheep with an obstructive thoracic oesophageal abscess. PMID- 8236645 TI - The veterinary profession and animal welfare. PMID- 8236646 TI - Oestrus control in ferrets. PMID- 8236647 TI - Pyometra and torsion of the uterus in a male dog. PMID- 8236648 TI - Feline bordetellosis: challenge and vaccine studies. AB - Four eight-week-old cats, shown to be free from feline calicivirus, feline herpesvirus and Chlamydia psittaci were challenged with an aerosol of Bordetella bronchiseptica. Within five days the cats developed signs of respiratory disease, characterised by nasal discharge, sneezing, spontaneous or induced coughing and dry or wet rales at auscultation. These signs were present for about 10 days, after which they began to resolve. To test the protective capacity of an experimental fimbrial antigen-based subunit vaccine, 10 kittens were vaccinated twice, with two weeks between the vaccinations, and five kittens were left unvaccinated. Two weeks after the booster the 15 kittens were challenged with an aerosol of B bronchiseptica as the sole pathogen. On the day of challenge the vaccinated kittens had a mean bordetella antibody titre of 2(9.5) whereas the control cats remained seronegative (titre < 2(2)). The control cats developed signs of respiratory disease after challenge, whereas the vaccinated cats were almost completely protected. The degrees of protection against rhinitis, sneezing, spontaneous or induced coughing, and dry or wet rales at auscultation were 100 per cent, 95 per cent, 95 per cent and 100 per cent, respectively. Furthermore, the vaccinated kittens cleared the challenge bacteria more quickly than the controls, resulting in a reduction of 80 per cent on days 15 and 18 after challenge and a reduction of 99 per cent on days 22 and 29 after challenge. The results show that B bronchiseptica can act as a primary pathogen in cats and that a vaccine containing the fimbrial antigen induces a protective immune response. PMID- 8236649 TI - Sensitivity, specificity and predictive value of blood cultures from cattle clinically suspected of bacterial endocarditis. AB - This study investigated the number of blood culture-positive cattle among 215 animals clinically suspected of having bacterial endocarditis. For animals that were necropsied, the sensitivity, specificity and predictive value of the diagnosis of endocarditis were calculated on the basis of the isolation of the causative bacteria from blood. Furthermore, it was investigated whether the glutaraldehyde coagulation time, total leucocyte count, per cent neutrophil granulocytes, pulse rate and duration of disease could help to discriminate endocarditis from other diseases. Among 138 animals necropsied the sensitivity, specificity and predictive value of blood cultivation were 70.7 per cent, 93.8 per cent and 89.1 per cent, respectively. None of the other measurements could be used to discriminate between endocarditis and non-endocarditis cases. PMID- 8236650 TI - Automated feeding of milk replacer and health control of group-housed veal calves. AB - The behaviour, growth rate, feed intake, health and slaughter quality of group housed veal calves fed automatically with milk replacer were compared with those of veal calves kept in individual crates and bucket-fed the same milk replacer. The growth rate and feed intakes of the group-housed calves were lower than those of the calves in individual crates. Their feed conversion rates were similar and there were no significant differences in carcase conformation. The meat of the group-housed calves was paler in colour. The feeding behaviour of the group housed calves was studied to see whether sick animals could be identified at an early stage but behaviour alone provided an inadequate detection system. Intensive health controls in one of the trials showed that infectious respiratory and digestive diseases may be a greater problem in group-housed veal calves than in veal calves kept in crates. PMID- 8236651 TI - Prophylactic efficacy of moxidectin for periparturient ewes and mid-summer lambs. PMID- 8236652 TI - Differentiation between enzootic and sporadic bovine leukosis by use of serological and virological methods. PMID- 8236653 TI - Hip dysplasia in a nine-month-old male Jersey calf. PMID- 8236654 TI - Pulmonary emphysema in an Arctic fox. PMID- 8236655 TI - Welfare implications of modern artificial breeding techniques for dairy cattle and sheep. AB - The legislation which protects animals from cruelty and poor welfare is described, from both a European and British perspective. Using well-established definitions of welfare, it can be seen that certain current practices related to farm animal breeding may constitute poor welfare particularly for dairy cattle. Taking only scientific measures of welfare as criteria for acceptable practice may not completely satisfy public concern over some biotechnical innovations in animal breeding. A reassessment of attitudes towards manipulation of animals for human use may be necessary. PMID- 8236656 TI - Long distance transport of lambs and the time needed for subsequent recovery. AB - The effects of nine and 14 hours of road transport and the subsequent recovery in lairage of 392 hill lambs were studied in August and November. The gathering and the handling of the lambs were stressful, both physically and psychologically, and the journey imposed further psychological and metabolic stress. The levels of noise in the trailer were high (90db[A]). There were no measurable differences between the responses of the lambs transported for nine or 14 hours and there appeared to be three stages in their recovery after transport. After the first 24 hours of lairage changes in the blood components usually associated with short term stress and dehydration had recovered; after 96 hours there had been a well defined recovery of liveweight and the levels of most of the metabolites measured appeared to have stabilised and after 144 hours the lambs had recovered almost completely, most of the creatine phosphokinase had been cleared from the plasma and their plasma protein levels had stabilised. PMID- 8236657 TI - Efficacy of recombinant bovine somatotrophin in the treatment of fat cow syndrome. AB - Recombinant bovine somatotrophin was used in addition to conventional therapy to treat a sample of post-calving cows in a herd suffering a high incidence of fat cow syndrome. Somatotrophin was also given to cows before they calved. There were no significant differences between the treated and untreated groups in the plasma concentrations of glucose, beta-hydroxybutyrate, albumin, total protein or calcium. Significant decreases in mean plasma urea concentration were observed in the pre-calving cows seven and 10 days after treatment with somatotrophin, and a significant decrease in mean plasma urea concentration was also recorded in post calving cows with the fat cow syndrome. There was no evidence that somatotrophin, with its many potent actions in cows with acetonemia, exacerbated clinical ketosis. The authors' subjective clinical impression was that somatotrophin was helpful in the treatment of cows with the fat cow syndrome. PMID- 8236658 TI - Can ground wheat form dough-balls in pigs' stomachs? PMID- 8236659 TI - Perirenal and intrapelvic pseudocysts associated with weight loss in a one-and-a half-year-old ram. PMID- 8236660 TI - Skin inflammation following triclabendazole treatment of dairy cows. PMID- 8236661 TI - Docking of dogs. PMID- 8236662 TI - Importing disease. PMID- 8236663 TI - Is unnecessary suffering avoidable? PMID- 8236664 TI - Unusual neoplasm in a dog. PMID- 8236665 TI - Experimental challenge of sheep 18 months after vaccination with a live (S48) Toxoplasma gondii vaccine. AB - Sheep can be immunised against toxoplasma abortion with a live vaccine composed of S48 tachyzoites. In this study the immunity induced in sheep was examined 18 months after vaccination. Seventy-three sheep were divided into three groups. Group 1 was vaccinated and 18 months later, when they were 90 days pregnant, the 27 sheep were challenged orally with Toxoplasma gondii oocysts. These ewes produced significantly more live lambs, after a significantly longer gestation period, than the 34 unvaccinated, challenged ewes in group 2. In addition there was less placental damage and fewer lambs were born with antibody to the parasite in the vaccinated group than in group 2. The 12 ewes in group 3 served as unvaccinated, unchallenged control animals. The degree of protection induced by the S48 tachyzoites was as good 18 months after vaccination as previously observed six months after vaccination. PMID- 8236666 TI - A comparison of topical, subcutaneous and oral administrations of ivermectin to reindeer. AB - The topical administration of ivermectin to adult reindeer hinds was evaluated by comparing its antiparasitic efficacy at a dose of 500 micrograms/kg bodyweight with that of oral administration at a dose of 200 micrograms/kg and subcutaneous administration at dosages of 20 and 200 micrograms/kg. The comparison included faecal examinations for nematode eggs and larvae and the counting of warbles and throat bots (Oedemagena tarandi and Cephenomyia trompe larvae). Weight changes and calf birth weights were recorded. All the treatments were 100 per cent effective against warbles and all except the low subcutaneous dose (20 micrograms/kg) against throat bots. The anti-nematodal efficacy of the topically applied ivermectin, the orally administered drug and the low subcutaneous dose was inferior to that of the subcutaneous dose of 200 micrograms/kg. There were no differences in weight changes between the groups. PMID- 8236667 TI - Winter dysentery diagnosed by farmers in dairy herds in central Sweden: incidence, clinical signs and protective immunity. AB - The occurrence of winter dysentery, diagnosed by farmers and defined as an outbreak of diarrhoea among at least 30 per cent of adult cattle in a herd, was monitored in 256 dairy herds in an area of central Sweden. The cumulative incidence of winter dysentery between April 1988 and March 1989 was 28.5 per cent. A typical outbreak lasted for one to two weeks and 74 per cent of the outbreaks occurred between November and January. A decrease in milk yield was reported in 90 per cent of the affected herds and the cows showed respiratory signs in 57 per cent of them. There was a significant (P < 0.05) association between the occurrence of fever and coughing. In 31 per cent of the outbreaks the farmer also noticed diarrhoea among the calves. One-third of the affected herds had experienced an outbreak within the previous four years and 18 per cent had at least one further outbreak during the following two years. There was a significantly (P < 0.05) lower disease score in herds that had had an outbreak within the previous four years than in herds which had had a less recent or no previous outbreak, indicating the development of temporary immunity to the causative agent. PMID- 8236668 TI - Time to loss of somatosensory evoked potentials and onset of changes in the spontaneous electroencephalogram of turkeys during gas stunning. AB - The times to the loss of somatosensory evoked potentials (SEPs) and the onset of suppressed and isoelectric electroencephalogram (EEG) were investigated in turkeys as they were stunned with gas mixtures consisting of one of three mixtures: (A) 30 per cent carbon dioxide and 60 per cent argon in air (2 per cent residual oxygen and 8 per cent residual nitrogen); (B) 90 per cent argon in air (2 per cent residual oxygen and 8 per cent residual nitrogen); (C) 65 per cent carbon dioxide in air (7 per cent residual oxygen and 28 per cent residual nitrogen). The time to the loss of SEPs, EEG suppression and the onset of an isoelectric EEG, respectively, were 22, 16 and 35 seconds in mixture A, 44, 41 and 101 seconds in mixture B, and 15, 15 and 67 seconds in mixture C. Stunning turkeys with mixture A or B would be suitable under commercial conditions. Mixture C, containing 65 per cent carbon dioxide in air, is considered on humanitarian grounds to be unacceptable for stunning turkeys owing to the pungency of the carbon dioxide at this concentration. PMID- 8236669 TI - Continued presence of phocine distemper virus in the Dutch Wadden Sea seal population. PMID- 8236670 TI - Seminal vesiculitis and epididymitis in an Anglo-Nubian buck. PMID- 8236671 TI - Characterisation of verocytotoxin-producing Escherichia coli isolated from pigs and cattle in northern Italy. PMID- 8236672 TI - Prevalence of lung abscesses and interstitial pneumonia in ovine oestrosis. PMID- 8236673 TI - CAE virus seroprevalence in a mixed goat herd. PMID- 8236674 TI - Treatment of wing tip oedema in raptors. PMID- 8236675 TI - Postoperative analgesia using phenylbutazone, flunixin or carprofen in horses. AB - Horses undergoing surgery were randomly assigned to one of three groups to receive phenylbutazone at 4 mg/kg (n = 72), flunixin at 1 mg/kg (n = 68) or carprofen at 0.7 mg/kg (n = 63) by slow intravenous injection at the end of surgery, just before they were disconnected from halothane. Pain was assessed by either of two resident surgical clinicians (who did not know which non-steroidal anti-inflammatory drug had been given) when the horses first stood up, two and four hours later and the next morning. If repeated doses of analgesic drugs were given the time was recorded and taken as an end point for the study. The presence or absence of side effects was also recorded. In the three groups there was no significant difference between the types of surgery performed, the numbers of horses requiring further analgesia or the pain scores at any time. In the horses needing further analgesia there was a significant difference in the time after surgery at which the further analgesia was given between those in the flunixin group, 12.8 +/- 4.3 hours (mean +/- sd) and those in the phenylbutazone group, 8.4 +/- 4.6 hours; the carprofen group had an intermediate interval of 11.7 +/- 6.9 hours. Significantly fewer of the horses that received butorphanol during surgery needed further analgesia than of those that did not receive any opioid. PMID- 8236676 TI - Transmission of bovine spongiform encephalopathy to sheep and goats. AB - Spongiform encephalopathy has been confirmed in both 'positive' and 'negative' lines of Cheviot sheep (selected for their differential response on experimental exposure to scrapie) after intracerebral injection or oral dosing with brain homogenate derived from cattle with bovine spongiform encephalopathy (BSE). With either challenge the incubation period of the disease ranged from 440 to 994 days in both lines of sheep. In a similar experiment, three Anglo-Nubian goats developed the disease 506 to 570 days after intracerebral infection with the same BSE homogenate, and two of three goats developed the disease 941 and 1501 days after oral dosing; the other goat and some sheep from each of the experimental groups remain alive 1720 days after exposure. This is the first report of the experimental transmission of BSE to sheep and goats. PMID- 8236677 TI - Relationship between the levels of antibodies to bovine viral diarrhoea virus in bulk tank milk and the prevalence of cows exposed to the virus. AB - A positive relationship was found between the prevalence of cows in a herd which were antibody-positive for bovine viral diarrhoea virus (BVDV) and the level of antibodies to the virus in bulk tank milk as determined by an indirect enzyme linked immunosorbent assay (ELISA). In herds with an ELISA absorbance value of up to 0.20 there were very few or no antibody-positive cows, whereas in herds with an ELISA absorbance value of at least 0.81, 87 to 100 per cent of the lactating cows were antibody-positive to BVDV. An analysis of the level of antibodies to BVDV in milk samples from Sweden and Finland showed that of 123 Swedish herds, 83.7 per cent had detectable antibodies to BVDV in their bulk milk whereas only 3.1 per cent of the 291 Finnish samples were antibody-positive. The incidence of BVDV infection in 105 herds in one area of Sweden was determined by analysing two samples of bulk tank milk taken one year apart. The infection had apparently occurred recently in five of these dairy herds. PMID- 8236678 TI - Infections with Babesia canis and Ehrlichia canis in dogs in Zimbabwe. AB - Of 105 dogs examined at a veterinary hospital in Harare, Zimbabwe, 52 per cent had antibodies reactive with Ehrlichia canis in indirect fluorescent antibody tests, 26 per cent had Babesia canis parasites in peripheral blood smears and 17 per cent had both infections. None of the dogs with serological evidence of ehrlichiosis had typical E canis morulae detectable in blood smears. The infections were regarded as incidental findings not readily related to the reasons for examination in 46 per cent of the dogs with antibodies to E canis and 17 per cent of the dogs with both infections. The most common laboratory abnormalities were anaemia and thrombocytopenia and the prevalence and severity of these in concurrent infections were intermediate to those found in individual infections. There were no pathognomonic clinical signs or laboratory abnormalities which could be used to distinguish between individual and concurrent infections. However, there was a significantly higher prevalence of non-regenerative anaemia in dogs with antibodies to E canis than in dogs with both infections. The prevalence of thrombocytopenia was significantly higher in dogs with babesiosis than in dogs with antibodies to E canis and the prevalence of hyperglobulinaemia was significantly higher in dogs with both infections than in dogs with antibodies to E canis. PMID- 8236679 TI - Anthelmintic dosing intervals for horses: comparison of three chemical groups. PMID- 8236680 TI - An outbreak of dermatophytosis in barking deer (Muntiacus muntjak). PMID- 8236681 TI - Veterinary nursing examinations. PMID- 8236682 TI - Veterinary nursing examinations. PMID- 8236683 TI - Dangerous Dogs Act. PMID- 8236684 TI - Docking of dogs. PMID- 8236685 TI - Veterinary histopathology. PMID- 8236686 TI - Bovine chlamydiosis in the United Kingdom. PMID- 8236687 TI - Study aims to keep horses running in the heat of Atlanta. PMID- 8236688 TI - Giardiasis in lambs at pasture. AB - Faecal samples from a group of lambs at pasture were screened at weekly intervals for nine weeks for the presence of Giardia species using a modified zinc sulphate flotation method. Fifty-nine of 86 lambs (68.6 per cent) excreted giardia cysts on one or more occasions. They were first detected at approximately three weeks of age and the highest incidence of excretion of cysts occurred when the lambs reached a mean age of 37 days. The lambs had diarrhoea but it was attributed to the presence of Eimeria ovinoidalis. PMID- 8236689 TI - Pharmacokinetics and clinical effects in dogs of a sustained-release formulation of theophylline. AB - Theophylline was administered orally in a sustained-release preparation at a dose rate of 20 mg/kg bodyweight to six beagle dogs. A mean (+/- se) maximum plasma concentration of 12.2 +/- 0.65 micrograms/ml was obtained 5.17 +/- 0.70 hours after administration and it was calculated to have a mean residence time of 9.09 hours. When given at 40 mg/kg bodyweight twice daily for five days the theophylline preparation was well tolerated, but, the dogs developed a marked tachycardia and hyperpnoea which were attributed to the high plasma concentrations (> 30 micrograms/ml) reached during this dosage regimen. PMID- 8236690 TI - Immuno-dot blot as a rapid diagnostic method for detection of chlamydial infection in koalas (Phasolarctos cinereus). AB - The sensitivity and specificity of an immunoscreening test for anti-chlamydial antibodies in koala (Phasolarctos cinereus) serum were determined after the adsorption of non-specific antibodies. The results of the test were compared with complement fixation tests, tissue culture, gene probe analysis and dot blot immunoscreening for host-borne chlamydial antigen. The immunoscreening test was the most sensitive test for the identification of chlamydial infection in koala serum samples, furthermore it was rapid, taking approximately 16 hours to complete, and inexpensive. However, for the assay of swab material from koalas, gene probe analysis remains the most sensitive method of detection of chlamydiae. PMID- 8236691 TI - Unusual mortality associated with poxvirus-like particles in frogs (Rana temporaria). PMID- 8236692 TI - High mortality associated with megabacteriosis of proventriculus and gizzard in ostrich chicks. PMID- 8236693 TI - Experimental infection of chicks with Enterococcus hirae. PMID- 8236694 TI - Conserving wildlife. PMID- 8236695 TI - Chicken anaemia agent in South Africa. PMID- 8236696 TI - Bovine mammillitis virus growth in human cells. PMID- 8236697 TI - Disease from the east: history repeats itself. PMID- 8236698 TI - Hereditary problems in Italian spinones. PMID- 8236699 TI - Veterinarians warned of a rise in sheep scab outbreaks. PMID- 8236700 TI - Norbrook Lecture. Mechanisms of pain and of its control. AB - I have tried to convey the notion that the pain system is a very flexible one. There is considerable excitement at present that we are beginning to understand the mechanisms whereby this flexibility results in clinical pain states; this must be a good prelude to trying to prevent or reverse them. But what is the relevance of all this to pain states in animals? There are still fundamental aspects about which we know too little. Detection of pain in animals is still difficult, and even when pain is present we often cannot tell how much it is affecting the animal, especially under conditions of chronic disease. We still do not know enough about the efficacy and kinetics of analgesic drugs in each species, a task for pharmacologists and clinicians alike. We know that sensitisation of nociceptive systems can occur after tissue trauma, as occurs during surgery, but we do not yet know whether this can be effectively prevented. Against this background of tasks yet to be undertaken, we can be optimistic that we are on the brink of understanding the triggers that alter pain sensitivity, that we shall soon know which mediators are involved, and that this will give rise to new strategies for the prevention as well as the alleviation of pain. There is indeed a period of excitement ahead. In all of this we must not lose sight of the ethical and welfare considerations of using animals for pain research.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236701 TI - Activity of morantel citrate against strains of benzimidazole-resistant nematodes of sheep in the United Kingdom. AB - The activity of morantel citrate (5.94 mg/kg base) was determined in laboratory tests against field isolates of benzimidazole-resistant nematodes. Its efficacies against adult and seven-day-old worms were 100 per cent and 100 per cent for Cooperia curticei, 95.1 per cent and 69.8 per cent for Haemonchus contortus and 100 per cent and 82 per cent for Ostertagia circumcincta. Morantel citrate was 100 per cent effective against benzimidazole-susceptible Nematodirus battus and Trichostrongylus vitrinus, and it reduced faecal egg counts by 97.9 per cent in sheep infected naturally with benzimidazole-resistant H contortus and O circumcincta. PMID- 8236702 TI - Effects of acepromazine, pethidine and atropine premedication on lower oesophageal sphincter pressure and barrier pressure in anaesthetised cats. AB - Combinations of acepromazine maleate, pethidine hydrochloride and atropine sulphate (0.05 mg/kg) or acepromazine maleate and pethidine hydrochloride and acepromazine maleate alone or atropine sulphate (0.1 mg/kg) alone were used to premedicate cats before they were anaesthetised with thiopentone, to investigate their effects on gastric pressure, lower oesophageal sphincter pressure and barrier pressure under anaesthesia. Manometric measurements were made by using a non-perfused manometric technique. The lower oesophageal sphincter pressure was lowest in the cats premedicated with atropine sulphate alone. The difference in barrier pressure between the atropine (0.1 mg/kg) and acepromazine treated cats was highly significant. The risk of gastro-oesophageal reflux appeared to be highest with atropine (0.1 mg/kg) if barrier pressure is used as an indicator of the likelihood of reflux. PMID- 8236703 TI - Effects of gonadotrophin releasing hormone administered 11 days after insemination on the pregnancy rates of cattle to the first and later services. AB - In a trial using 2530 recorded services of 1619 animals on 19 commercial dairy farms, alternate cows were injected intramuscularly with 10 micrograms buserelin, an analogue of gonadotrophin releasing hormone (GnRH), (Receptal; Hoechst Animal Health) 11 days after insemination. Pregnancy was diagnosed manually six to 12 weeks after the last service in all but three herds, for which calving data were collected. The pregnancy rates were calculated for control and treated cows, paired for calving to first service interval, parity and week of service. For 520 pairs of control and treated cows the pregnancy rates to first service were 50.6 per cent and 60 per cent, respectively (P < 0.01). For the second and subsequent services each cow was grouped according to its treatment or control status in the immediately preceding dioestrus period, in addition to its status on day 11 after service. As a result there were three treated groups: treated, treated; treated, control, and control, treated for comparison with the control, control group. For 136 pairs of control, control and control, treated cows the pregnancy rates to second service were 41.2 per cent and 54.4 per cent, respectively (P < 0.05), and for 67 similar pairs of cows the pregnancy rates to the third or later service were 23.9 per cent and 52.2 per cent, respectively (P < 0.001). For 40 pairs of control, control and treated, treated cows the pregnancy rates to the third or later service were 15.0 per cent and 45.0 per cent, respectively (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236704 TI - Relationship between herd calving to conception interval and culling rate for failure to conceive. AB - The factors affecting herd calving to conception intervals and culling rate are the interval to first service, the efficiency of heat detection, the pregnancy rate and the maximum number of oestrous cycles allowed to occur before the cow is considered to be so far into lactation that even if it does conceive, the length of the dry period and the delay to the next calving will make its production uneconomic. Depending on the fertility factor (the product of the average herd pregnancy rate and the oestrus detection rate) it is the maximum number of oestrous cycles, usually dictated by the length of the calving interval desired, that decides the outcome. A graphical representation of these mathematical relationships demonstrates methods for achieving different economic and other objectives. PMID- 8236705 TI - Serotyping and distribution of Listeria isolates from cases of ovine listeriosis. PMID- 8236706 TI - Isolation of bluetongue virus serotypes new to Indonesia from sentinel cattle in west Java. PMID- 8236707 TI - Anthrax and cowdriosis in an African elephant (Loxodonta africana). PMID- 8236708 TI - Docking of dogs. PMID- 8236709 TI - National Meat Hygiene Service. PMID- 8236710 TI - Untreated milk as a source of verotoxigenic E coli O157. PMID- 8236711 TI - Aleutian disease in the ferret. PMID- 8236712 TI - Association in bitches between breed, size, neutering and docking, and acquired urinary incontinence due to incompetence of the urethral sphincter mechanism. AB - The association in bitches between breed, size, neutering and docking, and acquired incompetence of the urethral sphincter mechanism was investigated. Observational studies were conducted on referred and first-opinion cases of hormonal urinary incontinence and on referred, confirmed cases of incompetence of the urethral sphincter mechanism. Large and giant breeds were at high risk, whereas small breeds were at low risk. Specific breeds at high risk were the old english sheep-dog, rottweiler, dobermann pinscher, weimaraner and Irish setter. The labrador retriever had a relatively low risk. There was also a positive association between docking and neutering, and the two conditions. PMID- 8236713 TI - An estimate of the prevalence of enzootic abortion of ewes in Scotland. AB - The prevalence of enzootic abortion of ewes in Scotland was estimated over a period of five years. Between 1987 and 1991, 30.7 per cent of all flocks with breeding ewes in Scotland submitted ovine abortion material, and of these flocks 28 per cent showed evidence of the infection, giving a prevalence of 8.6 per cent. Regional differences were observed in both the submission rate and the prevalence. PMID- 8236714 TI - Determination of the optimal treadmill slope for reproducing the same cardiac response in saddle horses as overground exercise conditions. AB - The purpose of this study was to define the most suitable treadmill slope for reproducing the same heart rate response as in horses being ridden on a track. Seven French saddle horses were exercised first on a level turf track and then on a treadmill. On the track the exercise test consisted of four periods of three minutes of increasing speeds at the trot and the gallop (96 to 600 m/minute). The treadmill exercise tests consisted of seven steps of increasing slope (0 to 9.6 per cent) at the trot (252 m/minute), followed, after an active recovery period at the walk with a 0 per cent slope, by five steps of increasing slope for two minutes each (0 to 6.3 per cent), at the gallop at 493 m/minute. The mean heart rate at each step of the exercise tests was measured with a heart rate recorder. On the exercise track there was a strong linear relationship (R = 0.89, P < 0.01) between heart rate and speed. The heart rate response of the horses exercised at different speeds and slopes on the treadmill was best fitted (R = 0.96, P < 0.01) by a multilinear model. The optimal treadmill slope was determined by equalizing the equations derived from the two tests: s = -0.001 V + 3.658. It was concluded that the optimal treadmill slope ranged between 3.0 and 3.7 per cent. In order to verify this result, a comparison was made between the heart rate response of the horses during the same incremental exercise test performed on the track and on the treadmill with a 3.5 per cent slope.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236715 TI - Hydrometra in dairy goats: reproductive performance after treatment with prostaglandins. AB - An intramuscular injection of 5 mg dinoprost caused a discharge of uterine fluid (cloudburst) in 49 cases of hydrometra in goats during the breeding seasons of 1988, 1989 and 1990. A spontaneous oestrus after the cloudburst was allowed to occur in 20 of the goats; in nine (45 per cent) of them a hydrometra recurred, three conceived at the first oestrus and eight returned to oestrus. Oestrus was induced in 29 other cases by means of a second intramuscular administration of 5 mg dinoprost, 12 days after the cloudburst. In this group a hydrometra recurred in only one goat, 14 goats (48 per cent) conceived at the first oestrus and 14 returned to oestrus. Of the animals in which a pseudopregnancy occurred once or more during the same breeding season, 85 per cent became pregnant, compared with 97 per cent of unaffected older goats. The mean number of kids of the goats that became pregnant and kidded after treatment for hydrometra was 2.0 compared with 2.3 for unaffected animals. The results indicate that a single administration of prostaglandin is not a satisfactory therapy for a hydrometra, but that reproductive performance improves when a second injection is given 12 days after the cloudburst. PMID- 8236716 TI - Osteitis and sequestrum formation of the interdental region of the mandible in 11 polo ponies. PMID- 8236717 TI - Persistence of African swine fever antibody reactivity on ELISA and immunoblotting assays. PMID- 8236718 TI - Technique of hoof biopsy in cattle. PMID- 8236719 TI - Detomidine-ketamine anaesthesia in chickens. PMID- 8236720 TI - The veterinary profession and animal welfare. PMID- 8236721 TI - National meat hygiene service. PMID- 8236722 TI - Crop stasis and regurgitation in racing pigeons. PMID- 8236723 TI - Behaviour of lame and normal dairy cows in cubicles and in a straw yard. AB - The behaviour of normal cows in cubicles was compared with that of normal cows in a straw yard and that of lame cows in cubicles. The normal cows in a straw yard lay down for longer in total (9.6 hours vs 6.8 hours) and during the night (8.55 hours vs 4.75 hours) and for significantly longer at a time (3.95 hours vs 2.45 hours) than normal cows in cubicles. The normal cows in a straw yard spent more time lying down and ruminating (5.1 hours) than normal cows in cubicles (3.3 hours). Lame cows in cubicles lay down for significantly longer during the day (3.3 hours) than normal cows in cubicles (2.1 hours). Although lameness did not affect the total time the cows spent in feeding and rumination, lame cows moved about less, and they adopted abnormal postures suggesting discomfort. PMID- 8236724 TI - Prevalence and incidence of scrapie in The Netherlands: a questionnaire survey. AB - Two independent nationwide surveys of randomly selected sheep farmers were conducted: in one a questionnaire was mailed to farmers and in the other the farmers were interviewed. The questionnaire was designed to obtain data on the occurrence of scrapie in the national sheep flock of the Netherlands. Farmers owning 30 or more ewes were selected from a database at the Ministry of Agriculture, Nature Management and Fisheries, resulting in 10,000 addresses. Two lists of names were compiled at random. In the postal survey 700 questionnaires were distributed, and 137 farmers were interviewed directly. The response rate in the postal survey was 37 per cent and in the interview survey 100 per cent. The results from the two surveys did not differ significantly. Six per cent of the farmers indicated that they had at some time observed signs of scrapie in their flock. For the total sheep population in the Netherlands, the prevalence of flocks with sheep showing signs of scrapie was between 3.8 per cent and 8.4 per cent (with 95 per cent certainty). In 1989 and 1990 the incidence rate within infected flocks was 1.27 cases/100 ewes/year, and the incidence rate for the total sample population was 0.1 case/100 ewes/year. PMID- 8236725 TI - Laboratory diagnosis of strangles and the isolation of atypical Streptococcus equi. PMID- 8236726 TI - Comparison of the efficacy of injectable and topical moxidectin for the reduction of faecal egg counts in cattle. PMID- 8236727 TI - A needle aspirate as an aid to diagnosis of lymphosarcoma in a guinea pig. PMID- 8236728 TI - Control of Johne's disease in a flock of sheep by vaccination. PMID- 8236729 TI - Symptoms and clinical pathology of rabies in the camel. PMID- 8236730 TI - Reducing the use of laboratory animals. PMID- 8236731 TI - Unusual neoplasm in a dog. PMID- 8236732 TI - Doramectin--a novel avermectin. PMID- 8236733 TI - Therapeutic and persistent efficacy of doramectin against Boophilus microplus in cattle. AB - One therapeutic and one persistent efficacy study were conducted in Brazil to evaluate doramectin at a dose rate of 200 micrograms kg-1 against induced infestations of the single host tick, Boophilus microplus. Doramectin was highly effective in eliminating established tick populations from cattle and also in preventing infestation by the parasite. In the therapeutic study, 12 calves were infested three times a week along the dorsal line with 2500 recently hatched larvae, for a total of 11 times before treatment. Animals were allocated to two groups on the basis of uniformity of established engorged tick burdens. Six calves were treated with doramectin and six received saline solution. From Day -3 to Day 21 post-treatment, individual collections of detached engorged female ticks were made from each calf. In the persistent efficacy study, 12 calves were allocated to two groups of six animals. Six calves were treated with doramectin and six received saline solution. From Day 1 to Day 17 post-treatment, each animal was infested three times a week along the dorsal line with 2500 recently hatched Boophilus microplus larvae, for a total of nine times. From Day 18 to Day 42 post-treatment, daily collections of detached engorged female ticks were made from individual animals. In the therapeutic study, efficacy (reduction of collected engorged female ticks) progressed from 51% at 24 h post-treatment (p.t.) to at least 99% at 4 days p.t., and reached 100% at 8 days p.t. With the exception of one tick that did not lay eggs, recovered from one animal at 11 days p.t., no more ticks were recovered from doramectin-treated calves for the duration of the experiment. For the first 6 days after treatment, only a few detached engorged ticks were collected from treated animals, and their oviposition and hatchability declined rapidly. In the persistent efficacy study, doramectin treatment was highly efficacious in preventing the establishment of Boophilus microplus populations for 20 days after the first ticks completed their cycle in the non-treated group. The oviposition and hatchability of the few ticks that completed their life cycle in the doramectin group were severely reduced. PMID- 8236734 TI - Effect of formulation on the pharmacokinetics and efficacy of doramectin. AB - The pharmacokinetics of doramectin, a novel avermectin, were evaluated following parenteral administration in a range of oil-based formulations in an attempt to optimise the formulation. Therapeutic and persistent efficacies against Cooperia oncophora were also evaluated. This approach led to the identification of formulations based upon sesame oil and ethyl oleate which gave more prolonged doramectin plasma concentrations with no loss in therapeutic efficacy and improved persistent efficacy following subcutaneous administration to cattle at a dosage of 200 micrograms kg-1. The importance of using both pharmacokinetic and efficacy end points to distinguish between formulations is discussed. All formulations were well tolerated as evidenced by the absence of any reaction to injection either in the form of behavioural responses, injection site swelling or postmortem lesions. Sesame oil with ethyl oleate was the best parenteral vehicle tested for doramectin, allowing the expression of a high level of therapeutic and persistent efficacy and offering the benefit of excellent injection site toleration. PMID- 8236735 TI - Activity of doramectin against nematode endoparasites of cattle. AB - A series of 28 controlled anthelmintic studies, involving 634 cattle, was conducted throughout North America and Europe to evaluate the efficacy of doramectin against a broad range of gastrointestinal parasitic nematode species and lungworms in naturally and experimentally infected animals. Within each study, one or two groups were treated with doramectin at 200 micrograms kg-1 and another group received no drug treatment. Worm burdens were estimated by standardised techniques, and efficacy assessed on reduction of worm burdens in doramectin-treated animals. Doramectin was at least 99.6% effective (P < 0.0002) in eliminating the immature and adult stages of the following 14 species of nematodes: Ostertagia ostertagi (including inhibited), Ostertagia lyrata, Haemonchus placei, Trichostrongylus axei, Trichostrongylus colubriformis, Cooperia oncophora (including inhibited), Cooperia pectinata, Cooperia punctata, Cooperia spatulata, Cooperia surnabada, Bunostomum phlebotomum, Strongyloides papillosus, Oesophagostomum radiatum and Dictyocaulus viviparus. Efficacy against adult Trichostrongylus longispicularis, Nematodirus spathiger and Trichuris spp. was 93.1%, 96.5% and 94.6%, respectively. Efficacies against adult and fourth stage larvae of Nematodirus helvetianus, the dose-limiting species, were 73.3% and 75.5%, respectively. PMID- 8236736 TI - Efficacy of doramectin against field nematode infections of cattle in Latin America. AB - Two studies were conducted, one in Argentina and one in Brazil, to evaluate the therapeutic efficacy of doramectin administered subcutaneously at a dose rate of 200 micrograms kg-1 (1 ml per 50 kg) to cattle harboring mixed field infections of gastrointestinal nematodes. In each trial, 20 heifers were randomly allocated to a control or treated group of ten animals each, on the basis of fecal egg counts. Animals in the treated and control groups received an injection of doramectin or saline in the lateral midline of the neck. From 14 to 18 days post treatment an equal number of animals from each group were slaughtered every day and the worm burdens of doramectin-treated animals were compared with those of non-treated controls. The efficacy of doramectin was at least 99.9% against adult stages of Ostertagia ostertagi, Haemonchus placei, Haemonchus contortus, Haemonchus similis, Trichostrongylus axei, Cooperia pectinata, Cooperia spatulata, Cooperia surnabada (Cooperia mcmasteri), Oesophagostomum radiatum and Dictyocaulus viviparus. Efficacy against Nematodirus helvetianus was 97.9% while efficacy against Trichuris discolor was 92.3%. Activity against inhibited larval forms of Ostertagia ostertagi, H. placei, Oesophagostomum radiatum, and Trichostrongylus axei, was at least 99.9%. PMID- 8236737 TI - Persistent efficacy of doramectin against experimental nematode infections in calves. AB - Three studies were conducted involving cattle exposed to experimental nematode infections. These studies were designed to investigate the prophylactic activity of a single subcutaneous treatment of doramectin at 200 micrograms kg-1 body weight against infections of Ostertagia ostertagi, Cooperia oncophora and Dictyocaulus viviparus. For each study, parasite-naive calves were randomly allocated to either a treated or a matched control group. One group received doramectin and the other received doramectin and the other received either no treatment or an injection of saline at 1 ml per 50 kg body weight by the subcutaneous route. Thereafter, all calves received a daily oral challenge of infective larvae of the particular parasite species on test in each study. Challenge of each pair of treatment/control groups continued for periods of 14, 21 or 28 days. An interval of 14-21 days was then allowed to permit the parasites which had established to mature, after which all animals were slaughtered and their worm burdens determined using standard techniques. Geometric mean worm burdens were calculated from the log worm counts and used to estimate percentage efficacy. Accumulated burdens of C. oncophora in doramectin-treated cattle resulting from a daily challenge infection for 14 or 21 days were reduced by 99.2% and 90.7% respectively, in comparison with those of non-treated control animals. For D. viviparus, burdens were reduced by 100% and 99.9% after a 21 or 28 day challenge, respectively. The corresponding figures for O. ostertagi were 99.9% after a 21 day challenge and 93.7% after a 28 day challenge. PMID- 8236738 TI - Doramectin--a potent novel endectocide. AB - Doramectin, 25-cyclohexyl-5-O-demethyl-25-de(l-methylpropyl)avermectin A1a, was selected as the best of a series of novel avermectins prepared by mutational biosynthesis. The primary evaluation of its in vivo antiparasitic activity was carried out using a rat Trichostrongylus colubriformis model and a rabbit Psoroptes cuniculi model. In each case the new avermectin performed favourably relative to dihydroavermectin B1a (DHAVM), the major component of ivermectin. Doramectin was extensively evaluated in cattle using an experimental micelle formulation, proving highly effective in cattle infected with Ostertagia ostertagi, Cooperia oncophora and Dictyocaulus viviparus when administered subcutaneously at 200 micrograms kg-1. The plasma pharmacokinetic characteristics of doramectin in cattle following intravenous administration revealed a plasma half-life of approximately 89 h. In the micelle formulation, doramectin administered subcutaneously at 400 micrograms kg-1 provided persistent activity against infection of cattle with C. oncophora and O. ostertagi for at least 8 and 12 days respectively. PMID- 8236739 TI - Efficacy of doramectin in the prevention of gastrointestinal nematode infections in grazing cattle. AB - Two studies were performed to investigate the efficacy of doramectin in the prevention of infection with Ostertagia ostertagi and Cooperia oncophora in grazing calves. In each study, 24 parasite-naive calves were randomly allotted to two equal groups and treated with either doramectin at 200 micrograms kg-1 or saline prior to mid-season turnout (Day 0) onto contaminated pasture. Faecal egg counts were carried out twice weekly from 15 to 64 days after turnout and the cumulative faecal egg count was calculated for each group of calves. In the doramectin-treated animals, eggs first appeared in the faeces 19 days and 22 days later than in controls for Studies 1 and 2, respectively. Mean cumulative faecal egg counts over the 64 days were reduced in the doramectin-treated groups by 71% and 87% for Studies 1 and 2, respectively (P < 0.01). The potential utility of injectable doramectin in the seasonal control of gastrointestinal nematode infestations in relation to these findings is discussed. PMID- 8236740 TI - Efficacy of doramectin against eyeworms (Thelazia spp.) in naturally and experimentally infected cattle. AB - The anthelmintic efficacy of doramectin was assessed for the control of Thelazia spp. in two studies using 44 naturally or experimentally infected calves. Cattle were randomly assigned based on weight to either a doramectin treatment group or a saline control group. Treated animals received doramectin at a dosage of 200 micrograms kg-1, subcutaneously in the lateral midline of the neck. Control animals received an equivalent dosage of 0.9% sterile saline. Animals were slaughtered 13-16 days after treatment and all eyes and associated tissues (including the lacrymal glands and ducts) were removed and examined for total number, species and viability of eyeworms. Two species of eyeworms, Thelazia skrjabini and Thelazia gulosa were found in the control group of naturally infected calves. Only T. skrjabini was found in the control group of experimentally infected calves. No eyeworms were found in any doramectin-treated animal. The efficacy of doramectin against Thelazia spp. in both naturally and experimentally infected calves was 100%. PMID- 8236741 TI - Spectrum of activity of doramectin against cattle mites and lice. AB - A series of 16 studies was conducted involving 288 cattle with naturally acquired or experimentally induced mite or louse infestations to evaluate the efficacy of doramectin. Four studies were conducted against the mite species Psoroptes bovis and two against the mite species Sarcoptes scabiei. Ten studies involved cattle with one or more of the following louse species: Damalinia bovis, Haematopinus eurysternus, Linognathus vituli and Solenopotes capillatus. In each study, a group of at least nine but not more than 25 cattle were randomly allocated to a control or a treated group. One group was treated with doramectin at 200 micrograms kg-1 and the other group received no medication. Mites or lice were quantified by species immediately before treatment and at weekly intervals for 4 weeks thereafter. Doramectin was 100% efficacious against infestations of P. bovis, Sarcoptes scabiei, H. eurysternus, L. vituli and Solenopotes capillatus. The drug reduced infestations of D. bovis by a mean of 82%. PMID- 8236742 TI - Efficacy of doramectin against warble fly larvae (Hypoderma bovis). AB - The efficacy of doramectin in the treatment of cattle harbouring naturally acquired infestations of first, second and third instar larvae of Hypoderma bovis was determined in two studies carried out in the Burgundy region of France. In the first study, cattle on six farms with a history of H. bovis infestations were treated during October 1989 with either doramectin at a dose of 200 micrograms kg 1 liveweight (186 animals) or with an equivalent volume of saline (157 animals). During the following March and April, all animals were examined for the presence of warbles. In the second study, cattle on four farms with warbles present in their backs were treated during March 1990, with either doramectin or saline (as before). The viability of larvae within each warble on all the animals was then assessed every 2 days for 14 days. In Study 1, no warbles were present in any of the doramectin-treated cattle at any time, whereas warbles were found in 135 saline-treated animals. In Study 2, all larvae in warbles on the backs of the doramectin-treated cattle were dead by Day 14 after treatment, whereas viable larvae were still present in warbles in the backs of all saline-treated cattle. No adverse reaction to doramectin treatment was observed in any animal at any time. It was concluded that doramectin is both safe and 100% efficacious in the treatment of first, second and third instar H. bovis infections of cattle. PMID- 8236743 TI - Therapeutic and persistent efficacy of doramectin against Dermatobia hominis in cattle. AB - One therapeutic and one persistent efficacy study were conducted in Brazil to evaluate doramectin at a dose rate of 200 micrograms/kg-1 against induced infestations of the tropical warble-fly, Dermatobia hominis. Doramectin was very effective in both the treatment of established infestations and also in the prevention of damage caused by the parasite. In the therapeutic trial, 12 calves were infested along the dorsal line with 25 first instar larvae of recent field isolates of D. hominis but in one calf nodules did not develop. Twenty-four days later animals were allocated to two groups on the basis of the number of parasite nodules present. Six calves were treated with doramectin, and five received saline solution. Animals were examined daily for 11 days post-treatment and the number of nodules mapped and recorded. Larvae that completed development were collected and incubated to evaluate viability. In the persistent efficacy study, 24 calves were allocated to six groups (T1-T6) of four animals each. On the day of treatment, three groups (T1, T3 and T5) were treated with saline and three groups (T2, T4 and T6) with doramectin. At 21 days, 28 days and 35 days post treatment, 25 first instar D. hominis larvae were seeded along the dorsal line of each calf of T1 and T2, T3 and T4, and T5 and T6, respectively. Animals were examined daily for 18 days and the number of nodules mapped and recorded 6, 12 and 18 days post-infestation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8236744 TI - Prophylactic and persistent efficacy of doramectin against Cochliomyia hominivorax in cattle. AB - Two prophylactic studies and one persistent efficacy study were conducted in Brazil to evaluate one injection of doramectin at a dose rate of 200 micrograms kg-1 against induced infestations of the New World screwworm, Cochliomyia hominivorax. In each prophylactic study, 12 calves were allocated to two groups of six animals. Six calves were treated with doramectin and six with saline. Two hours after treatment, animals were anesthetized and received two incisions, one in the shoulder and one in the rump. Each incision was then infested with 50 first instar larvae. In the persistent efficacy study, 24 calves were allocated to six groups (T1-T6) of four animals each. On the day of treatment, three groups (T1, T3 and T5) were treated with saline and three groups (T2, T4 and T6) with doramectin. All animals were anesthetized and received four incisions, one in the shoulder and one in the rump of each side. Each incision was then infested with 30 first instar larvae of C. hominivorax with the following schedule: T1 and T2 at Day 3, T3 and T4 at Day 7, and T5 and T6 at Day 14 post-treatment. Calves were observed daily and incisions evaluated for 8 days post-infection. Larvae that completed development and exited the wounds were collected and incubated to evaluate viability. In all experiments, doramectin was 100% efficacious in preventing screwworm infestation caused by field isolates of C. hominivorax. First instar larvae were eliminated from the incisions of doramectin-treated calves between 24 and 48 h post-treatment, and no third instar were recovered at any time. As a consequence, healing started at 24 h and was completed at 96-120 h post-infection. Screwworm infestations and viable third-instar larvae developed in all the incisions in non-treated calves, and healing did not begin until larvae had completed their normal life cycle and exited the wounds. The persistent efficacy of a single injection of doramectin extended beyond 14 days. PMID- 8236745 TI - Evaluation of bone-wax extract on the frequency of chromosomal aberrations in the bone marrow of Swiss mice. AB - Indigenously developed bone wax was evaluated for cytogenetic effects of a saline extract of the bone wax. Swiss mice were assigned to 5 groups of 6 animals each. Groups I, II, III were given non-lethal doses of 2.5, 5.0 or 10.0 ml bone-wax saline extract/kg body wt and Groups IV and V received sterile 0.9% saline vehicle or cyclophosphamide ip as controls. After 24 h the mice were sacrificed by cervical dislocation. Bone marrow preparations were stained with Giemsa's stain and examined by light microscopy. Abnormalities such as numbers of chromosome gaps, breaks and translocations were noted. The bone-wax saline extract did not induce chromosomal aberrations in the bone marrow of Swiss mice under our laboratory conditions. PMID- 8236746 TI - Studies of sheep experimentally poisoned with Hypericum perforatum. AB - Sheep given different dosages and frequencies of Hypericum perforatum had decreased hemoglobin, red blood cell count, packed cell volumes, total protein, glucose, cholesterol, triglycerides, and serum alkaline phosphotase activities. Blood urea nitrogen, sodium, potassium, bilrubin (total and direct), and the activities of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase and gamma glutamyltransferase increased. Hemato-biochemical assays were useful diagnostic aids to determine the severity of this plant's toxic effects. PMID- 8236747 TI - A bird's eye review of the evolution of the Canadian Poison Control Program (1957 1989). PMID- 8236748 TI - A survey into the perceived usefulness of the NPIS poster "A guide to commonly ingested non-toxic substances". AB - A poster listing non-toxic substances was distributed to all accident and emergency departments in the UK by the London center of the National Poisons Information Service (NPIS). To assess the usefulness of the poster and its impact upon the information seeking behaviour of NPIS users, a survey was carried out by postal questionnaire. This is the first such user survey carried out by the NPIS. A total of 197 questionnaires were returned (71%). Of those respondents who had received the poster, 123 (70%) indicated they would use the poster instead of contacting the NPIS. A larger number (169 or 96%) indicated they found the poster useful; in these cases it was possible the poster would be used as a training tool. A total of 74 (39%) respondents had a policy of contacting the NPIS for every case of potential poisoning, and of these only 21 indicated that they would not use the poster instead of contacting the NPIS. This survey highlights the potential benefits of such projects for both poisons information services and their users, and the need for a full NPIS user survey in order that future developments are readily incorporated into existing working practices. PMID- 8236749 TI - Microbiological significance of drug residues in food: welcome and introduction. PMID- 8236750 TI - Symposium. Microbiological significance of drug residues in food. June 8-9, 1992, Rockville, Maryland. PMID- 8236751 TI - Microbiological significance of drug residues in food: symposium objectives. PMID- 8236752 TI - The normal intestinal microflora: ecology, variability and stability. AB - The composition and functions of the intestinal floras from humans and other animals is compared and contrasted. Intrinsic and extrinsic factors, such as anatomy, age, and diet, help define the nature of the flora typical to each species of animal. Stable differences, such as the ability to reduce cholesterol or not, between the flora of individuals of the same species are described. The difficulties of characterizing a flora by cultural taxonomy are discussed and alternatives are suggested. These alternatives, collectively known as microflora associated characteristics (MACs), are measures of the biochemical activity of the bacterial flora. How MACs can be used to measure the effects of subtherapeutic levels of antibiotics on a flora is described. PMID- 8236753 TI - Historical perspectives on regulatory issues of antimicrobial resistance. PMID- 8236754 TI - Clinical importance and public health concerns for the emergence of antibiotic resistant organisms in the human intestinal microflora. AB - Infections with antibiotic-resistant organisms clearly have an economic impact, although the extent remains uncertain. Major problems are: the problem of multiresistance, i.e., the mutation or rearrangement of a single gene to cause resistance to multiple antibiotics of several different classes at once; the linkage between virulence and resistance, and the influence of antibiotics on these two seemingly unrelated events; and the problem and consequences of gene dissemination. There are simply no data on antibiotic usage levels required for selection of some of these genetic events. This an area where much more information is needed. PMID- 8236755 TI - Human studies to measure the effect of antibiotic residues. AB - This epidemiological study compares the frequency of resistant bacteria in stool microflora among vegetarians and nonvegetarians over a 12 month period. Two well characterized vegetarian populations (one in Boston, MA and the other in Loma Linda, CA) as well as appropriate controls were studied. No apparent differences in the prevalence of antibiotic resistance in the microflora were noted; however, vegetarians had a significantly greater incidence of multi-antibiotic resistance. E. coli of the same API biotype had the same frequency of antibiotic resistance in both vegetarians and nonvegetarians. Quantitative studies showed similar percents of tetracycline resistant facultative isolates and of "bacteroides." Klebsiella were more common in the stool of the nonvegetarians. As shown in previous studies, exposure to animal products either as meat eaters or production workers in a poultry abattoir was not associated with an increased incidence of resistant bacterial flora or infections caused by resistant strains. PMID- 8236756 TI - Current status of models for testing antibiotic residues. AB - Models for testing the effect of antibiotic residues on the human gut flora should be (i) feasible (quality of data, statistics, price, duration), (ii) sensitive (baseline resistance, background variability, response to low doses), and (iii relevant (composition and ecology of the flora, predictive value). An evaluation of existing models led to the following conclusions: 1. Flora. Relevant models must include anaerobic flora that antagonize colonization by minor bacterial populations. In vitro tests on pure strains, batch or continuous flow cultures, do not take this barrier effect into account. 2. Drug treatment. Treatment should be long enough to allow the detection of indirect selection due to a weakening of the barrier effect (i.e., greater than 15 days). For dose response analysis, trials should include at least 3 groups: a control group, an experimental group given the antibiotic at residue levels, and a positive control group given a high dose of the same drug. 3. Target strain(s). The target bacteria should be identified, resident in the gut, accurately countable, drug susceptible but with drug resistant variants, and related to a pathogen. Some gut aerobes meet these requirements: sensitive to the ecosystem balance, easy to count accurately, opportunistic pathogens, and R-plasmid carriers. Total anaerobes are not suitable. An anaerobe species may serve as a target strain if it is particularly susceptible to, or selected by, the test antibiotic. Thus, each in vivo protocol should be based on in vitro data. 4. Measure of resistance. The determination of the minimum inhibitory concentration (MIC) of selected clones from the model flora is time consuming, does not detect subdominant resistance (less than 1%); and a MIC shift is difficult to test statistically. In contrast, direct counting of bacteria on drug supplemented media allows for the rapid measure of minor resistant populations. 5. Statistics. Most published designs do not include a power calculation, nor yield enough data points for accurate statistical analysis. Sufficient statistical power is critical for studies using conventional humans and animals, because the data are highly variable and not normally distributed. 6. Human trials. The lowest level of antibiotic tested in human volunteers (2 mg oxytetracycline/day for 7 days in 6 subjects) increased the proportion of resistant fecal enterobacteria (p = 0.05). However, the large day-to-day and inter individual variation in the human flora limit the significance of this change. Confirmation of this finding would require a large number of volunteers.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8236757 TI - European perspectives on the regulation of antimicrobial drugs. AB - EEC legislation has been drawn up over the past few years to harmonize requirements for authorizing and marketing veterinary medicines. These measures have taken into account not only antimicrobials used for treatment and prevention of disease but also those used for growth promotion. Existing Directives are in the process of amendment and in some instances the amended legislation has already been agreed upon. One of the issues dealt with in the original Directives was that of multiple antibiotic resistance, although only limited emphasis was placed on dealing with this aspect of microbiological concern. More recently, a Regulation has been issued which requires that an MRL (maximum residue level) should be set before Member States may grant marketing authorizations for any new product. A number of toxicological and pharmacological issues have to be addressed by the applicant in making up a dossier for determining the MRL, and the microbiological potential of residues is a subject which has to be considered. The applicant is given some degree of choice in determining precisely which studies to use to ascertain whether the induction or selection of resistant bacteria could alter the consumer's intestinal flora. This paper describes some of the background to the EC legislation on the studies required for residues and evaluates the need for such studies. PMID- 8236758 TI - The IR-4 Project. A national program for the approval of new animal drugs for minor species and minor uses. Accomplishments and goal for minor ruminants. PMID- 8236759 TI - An overview of current programs on drug development and regulation for small ruminants--pharmaceutical industry perspective. PMID- 8236760 TI - Drug needs of the sheep industry. AB - Two areas of drug needs in the sheep industry are reviewed. The first is the need for prolificacy drugs such as those used outside the country. These drugs would encourage out-of-season lambing, increased lambs/per ewe during the traditional lambing season and synchronize estrus for easier management of lambing and increased use of artificial insemination for faster genetic improvement. The other area is drugs to treat or prevent major causes of disease, specifically abortion, neonatal diarrhea, pneumonia and internal parasitism. Efficacious drugs for these diseases are either not now cleared for use in sheep at all; a safe, easy to administer product is not available; or not cleared for sheep at a dosage level which is effective. These are drugs which will have a positive impact on the industry by increasing production, reducing expenses, and helping the U. S. to be able to compete with other sheep producing countries in the U. S. markets. PMID- 8236761 TI - An overview of selected diseases and drug needs for deer. PMID- 8236762 TI - An overview of the drug needs and present lack of approved drugs for sheep, goats, llamas, deer and bison. PMID- 8236763 TI - Meat/hair goats: overview of diseases and the need for drugs. PMID- 8236764 TI - Special-fed veal calves: overview of production methods and drug needs. AB - Current American special-fed veal production methods and the results of a concerted and effective quality assurance effort are reviewed. The veal industry has been successful in markedly reducing the frequency of violative chemical concentrations (including antibiotics) according to Food Safety and Inspection Service (FSIS) monitoring programs. The relation of drug availability and animal well-being is discussed. The need for increased availability of safe and effective treatments, primarily for enteric and respiratory use, is emphasized by the large number of source dairy farms and marketing facilities represented within a group of veal (or dairy-beef) calves started by a producer. The veal calf is a preruminant and as such is not covered by calf/cattle labels; effects of animal age and diet on drug metabolism and excretion are discussed. The greatest need for drugs is during the first 4 to 6 weeks of the 16- to 18-week production cycle, allowing a potential 10- to 14-week withdrawal period for drugs used early in the production cycle. The drugs most frequently used in the veal industry under Extra-Label Drug Use (ELDU) provisions and valid Veterinarian/Client/Patient Relationships (VCPR), most of which have calf/cattle labels are listed. PMID- 8236765 TI - Major infectious and non-infectious diseases of the llama and alpaca. PMID- 8236766 TI - Drug development for small ruminants: reflections on current status. PMID- 8236767 TI - Consideration of sheep as a minor species: comparison of drug metabolism and disposition with other domestic ruminants. AB - The processes which determine bioequivalence of a given drug among species are many, and include absorption from a site of administration, renal, biliary and intestinal elimination, sequestration (in particular, binding to proteins or other macromolecules), distribution and redistribution, biotransformation, and receptor population density and uniqueness. The present review is limited to physiologic and pharmacologic parameters which affect drug distribution, elimination, and metabolism, primarily because these are areas where sufficient data is available to make comparisons between sheep and other ruminants. The literature suggest a high degree of similarity among domestic ruminants in the distribution and elimination of drugs that are not metabolized but eliminated by passive processes such as renal glomerular filtration. Although the data are not as uniform as when conducted under rigorous control in the same laboratory setting, it is possible to predict very similar pharmacokinetic profiles for many of the antimicrobials between cattle, sheep and goats. Metabolic scaling of kinetic parameters could be predicted in comparative studies conducted under well controlled conditions, in which case the greatest similarities would occur among the small ruminants with values being somewhat dissimilar between sheep or goats and cattle. Plasma protein binding of drugs appears to be very similar among the ruminants, and its influence on drug distribution and elimination does not appear to vary appreciably between cattle and sheep. There is, however, very little definitive data which describes binding parameters, and there is very little data of any kind on binding to caprine albumin other plasma proteins of the goat. Sheep apparently differ from cattle, however, in transcortin concentrations, and this could affect the distribution and elimination of prednisolone or any other synthetic steroid which exhibits high affinity binding to this transport protein. Among the drug substances for which comparative information is available, there is very little to suggest qualitative differences in routes of metabolism among ruminants. There appears to be a remarkable degree of similarity in both major and minor pathways of drug metabolism among these species, and in fact, no documentable differences of a qualitative nature have been found. It should be noted, however, that documentable differences would only be established in a direct comparison of species conducted in the same laboratory under carefully controlled conditions. Such conditions would necessarily include assurance of no recent exposure to enzyme inducing agents or inhibitors or to agents which could deplete endogenous substances necessary to the enzymatic process.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8236768 TI - Pharmaceutical tools for wildlife medicine and management: 2000 and beyond. AB - Pharmaceuticals will play an increasing role in wildlife management in North American in the future. Pharmaceuticals for use in wildlife medicine and management must be made available to the wildlife veterinarian and wildlife manager to address the situations existing today. The challenges for pharmaceuticals to be used in wildlife are 1) development of new technology and molecules, 2) acceptable route of delivery, and 3) the challenge of the federal regulatory process. All three aspects must come together in an environment of informed cooperation between the needs of the wildlife veterinarian, the pharmaceutical industry and the appropriate regulatory agencies. It is the collective responsibility of all three to ensure these essential tools are available to meet the challenges for wildlife pharmaceuticals beyond 2000. PMID- 8236769 TI - Evaluation of the efficacy and safety of morantel tartrate in domestic goats. PMID- 8236770 TI - Residue depletion of morantel tartrate in goat tissue and milk. PMID- 8236771 TI - Scrapie control in the United States. PMID- 8236772 TI - A review of feline infectious peritonitis virus: molecular biology, immunopathogenesis, clinical aspects, and vaccination. AB - Feline infectious peritonitis (FIP) has been an elusive and frustrating problem for veterinary practitioners and cat breeders for many years. Over the last several years, reports have begun to elucidate aspects of the molecular biology of the causal virus (FIPV). These papers complement a rapidly growing base of knowledge concerning the molecular organization and replication of coronaviruses in general. The fascinating immunopathogenesis of FIPV infection and the virus' interaction with macrophages has also been the subject of several recent papers. It is now clear that FIPV may be of interest to scientists other than veterinary virologists since its pathogenesis may provide a useful model system for other viruses whose infectivity is enhanced in the presence of virus-specific antibody. With these advances and the recent release of the first commercially-available FIPV vaccine, it is appropriate to review what is known about the organization and replication of coronaviruses and the pathogenesis of FIPV infection. PMID- 8236773 TI - A gelatin test to detect activity and stability of proteases produced by Dichelobacter (Bacteroides) nodosus. AB - Previously reported tests to distinguish thermostable and thermolabile proteases of Dichelobacter nodosus used hide powder azure as a test substrate. This paper describes an alternative test for protease stability using gelatin, an inexpensive and convenient substrate. The test required less equipment and time than the hide powder tests, and simplified the testing of multiple samples. Proteases from 2965 isolates of D. nodosus from samples collected as part of a footrot eradication scheme were tested using the gelatin method, 1707 produced thermostable, and 1258 produced thermolabile protease. Production of thermostable protease was used to identify isolates of D. nodosus which had the potential to cause the virulent form of ovine footrot. Comparisons were made with a hide powder test on 47 isolates. Further characterisation of all proteases was undertaken using polyacrylamide gel electrophoresis. Isoenzyme patterns of thermostable protease producing isolates were identified as S1 (1688 isolates) and S2 (19) whilst thermolabile protease producing isolates showed patterns U1 (1104 isolates), U2 (40), U3 (32), U4 (47), U5 (6), U6 (28) and S1 (1). PMID- 8236774 TI - Outer membrane proteins of three pathogenic Leptospira species. AB - The outer membrane proteins of seven reference strains of pathogenic Leptospira (L. alstoni serovar grippotyphosa, L. borgpetersenii serovar hardjo, and L. interrogans serovars autumnalis, bratislava, canicola, icterohaemorrhagiae, and pomona) were investigated to identify common surface-exposed outer membrane proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of sodium-N lauroylsarcosinate-insoluble outer membrane enriched fractions of the reference serovars and two field isolates of serovars hardjo and pomona revealed six common protein bands with approximate molecular masses of 77, 66, 42, 35.5, 24, and 18 kDa. At times the 35.5 kDa endoflagellar band resolved into two distinct bands, 35.5 kDa and 34 kDa. Immunoblotting of the same fractions using rabbit leptospiral antibodies showed six bands to be common (66, 59.5, 44, 42, 35.5, and 18 kDa). The 44 kDa band stained poorly with Coomassie blue but prominently by immunoblotting. Four reference strains (serovars bratislava, canicola, icterohaemorrhagiae, pomona), and two field isolates of serovar pomona and one of serovar bratislava were grown in low iron media to which the iron chelators 2,2' dipyridyl or ethylenediaminehydroxyphenylacetic acid were added. No iron dependent expression of outer membrane proteins was observed. The only difference observed between the outer membrane proteins when reference serovars of canicola or pomona were grown in dialysis bags in the peritoneum of swine or in vitro was the loss of the 77 kDa band from in vivo grown organisms. Treatment of whole leptospires with proteinase K did not remove the 77, 66, 59.5, or 42 kDa protein; these proteins may not be surface expressed or are inaccessible to the proteinase K. The 44 kDa band could not be evaluated by this method and the 18 kDa band was proteinase K resistant. PMID- 8236775 TI - Development of neutralizing antibody in horses infected with Ehrlichia risticii. AB - The role of the humoral immune response in ehrlichial infection is unknown. Development of neutralizing antibodies during a course of Ehrlichia risticii infection in a pony was examined in vitro by determining the inhibition of E. risticii infection of P388D1 cells in the presence of the sera. The pony experimentally infected with E. risticii developed significant neutralizing activity in the sera by 15 days postinfection when parasitemia started to decline. Neutralizing activity continued to rise after recovery from the disease up to 34 days postinfection at which time the experiment was terminated. In vitro neutralizing activities in the sera from 3 additional ponies infected with E. risticii were lower at 2 weeks than at 4 weeks postinfection. The sera from vaccinated/challenged ponies had comparable neutralizing activity to those of the recovered ponies at approximately 3 to 4 weeks postchallenge. Equine sera from infected or vaccinated/challenged ponies were also effective in protecting mice from E. risticii infection. These studies demonstrated the significant development of neutralizing activity in the sera of recovered or vaccinated/challenged ponies. PMID- 8236776 TI - Characteristics of Vero cytotoxin producing Escherichia coli associated with intestinal colonization and diarrhea in calves. AB - Isolates of Escherichia coli which produce Vero cytotoxin (VTEC) were obtained during 1983-1989 from calves raised in 5 north-central states of the USA. All of the calves experienced intestinal epithelial colonization by VTEC, diarrhea or both; twelve of the calves had bloody diarrhea. Twenty one isolates were serogroup O111 and the others were O103, O69, O45, 026, O5, or non-typable (4 isolates). All but one of the isolates hybridized with the CVD419 probe which identifies most VTEC strains. Thirty two isolates hybridized with the VT1 probe, 3 with both the VT1 and VT2 probes, and one with neither probe. The culture filtrate of the VT probe negative isolate was partially neutralized by SLT I monoclonal antibody. For the other isolates, the results of toxin neutralization by anti-SLT I and anti-SLT II monoclonal antibodies corresponded exactly with the VT1 and VT2 probe hybridization results. Three of the strains adhered in a localized manner to HEp-2 cells and Intestine 407 cells. PMID- 8236777 TI - Cross-reactivity between B. burgdorferi and other spirochetes affects specificity of serotests for detection of antibodies to the Lyme disease agent in dogs. AB - Western immunoblots, the kinetics-based enzyme-linked immunosorbent assay (KELA), and the microagglutination test were used to evaluate cross-reactivity among antibodies to serovars of Leptospira interrogans (leptospiral serovars), and B. burgdorferi from naturally infected dogs, and to Serpulina (Treponema) hyodysenteriae from vaccinated rabbits. Whole-cell lysates from Borrelia spp., leptospiral serovars, and Serpulina spp. were used for SDS-PAGE, western blots, and KELA. Crossreactivity occurred between the antibodies to B. burgdorferi and leptospiral serovars when tested on the heterologous antigens. Antibodies to leptospiral serovars tended to cross-react more strongly with antigens of B. burgdorferi spp. than did antibodies to B. burgdorferi when tested against antigens of leptospiral serovars. The antibodies against B. burgdorferi showed a lesser degree of cross-reactivity to the antigens of S. hyodysenteriae and S. innocens than they did to leptospiral serovars. We conclude that cross-reactivity occurs between B. burgdorferi and leptospiral serovars. Validation and interpretation of ELISA tests for detection of antibody activity to whole cell lysates of the Lyme agent must take this cross-reactivity into consideration. Conversely, dogs infected with the Lyme agent do not show significant cross reactivity in the microagglutination test for antibody to the leptospiral serovars. PMID- 8236778 TI - Comparison of a cold enrichment method and the IDF method for isolation of Listeria monocytogenes from animal autopsy material. AB - The method most often used in Sweden for isolation of Listeria monocytogenes from animal autopsy material is a cold enrichment method. This method is very slow. The International Dairy Federation (IDF) has recently presented a method for detection of L. monocytogenes in milk and milk products that is complete in one week. During a two year period 69 specimens from dead animals with suspected listeriosis were examined for L. monocytogenes in parallel analyses with both the cold enrichment method and the IDF method. Samples derived from different autopsy material representing a variety of animals. L. monocytogenes was isolated in 27.5% of the samples with the IDF method but only in 4.3% with the cold enrichment method. It is concluded that the IDF method was more sensitive than the cold enrichment method. PMID- 8236779 TI - Meningitis in pigs caused by Streptococcus suis--a speculative review. PMID- 8236780 TI - Detection and identification of equine herpesvirus-1 and -4 by polymerase chain reaction. AB - A rapid method for detection and identification of equine herpesvirus-1 and -4 (EHV-1 and EHV-4) was developed using polymerase chain reaction (PCR). Primers for PCR were designed from aligned nucleotide sequences of glycoprotein B genes of EHV-1 and EHV-4 to amplify specific regions for EHV-1 or EHV-4 or a common region of both viruses. By using type specific primer mixture, amplified fragments were identified as EHV-1 or EHV-4 in a one-step reaction. We have applied this technique on specimens from aborted fetuses. The samples contained only EHV-1 and there was complete accordance between the results of PCR and virus isolation. Our PCR system could differentiate the two virus types rapidly in a one-step reaction. PMID- 8236781 TI - Comparative hybridization and nucleotide sequence information from two noncytopathic isolates of bovine viral diarrhea virus. AB - Comparative hybridizations of 8 cytopathic (CP) and 11 noncytopathic (NCP) isolates of bovine viral diarrhea virus (BVDV) were done using 4 different cDNA hybridization probes. The hybridization probes were prepared from cDNA synthesized from 1 CP BVDV (NADL) and 2 NCP BVDV isolates (SD-1 and NY-1) within the p80 region and from cDNA spanning the 5' untranslated region of NCP SD-1. Hybridization with the 5'/SD-1 probe detected 19 out of 19 isolates, whereas the p80/NADL, p80/NY-1 and p80/SD-1 hybridization probes detected only 12, 16 and 13 isolates, respectively. To determine the basis for the different patterns of hybridization, the nucleotide sequence was determined for the p80/NY-1. The nucleotide sequence was compared with the published CP NADL and CP Osloss and NCP SD-1 and NCP1 nucleotide sequences and in 45% of the base differences between NY 1 and NADL, NY-1 and Osloss were divergent from NADL, SD-1 and NCP1. Based on comparative nucleotide sequence data and the different reactivities of the p80/NADL, p80/NY-1 and p80/SD-1 hybridization probes the relationships of the various test isolates were characterized. PMID- 8236782 TI - Role of pseudorabies virus glycoprotein II in protection from lethal infection. AB - A monoclonal antibody (mAb), named 1.21, with complement-dependent neutralizing activity was produced against glycoprotein II (gII) of pseudorabies virus (PRV). By immunoaffinity chromatography using a mAB 1.21 column, gII was purified from Nonidet P40-lysates of PRV infected BHK21/13 cells. When mice and pigs were immunized with purified gII, complement-dependent virus-neutralizing antibodies were produced. The immunized animals survived potentially lethal challenge with PRV. These results indicate that an immunological response against gII plays an important role in the protection from PRV infection. PMID- 8236783 TI - Detection of Mycoplasma meleagridis by polymerase chain reaction. AB - A pair of 32 base primers was synthesized based on the DNA sequence data of a Mycoplasma meleagridis (MM) species-specific recombinant, pMM-2. The primers were used in a MM-polymerase chain reaction (PCR) to amplify a target DNA of approximately 850 bp. Annealing temperatures ranging from 58 degrees C to 61 degrees C could be used for the MM-PCR without loss of specificity. The primers amplified 1 ng of DNA from 17 strains of MM, but not 10 ng of DNA from 16 heterologous species of avian mycoplasma, pUC8 plasmid, lambda phage or calf thymus DNA. The minimum amount of target DNA detected by MM-PCR was 10 fg, which indicated that this procedure was 100000 times more sensitive than dot blot methodology using an MM recombinant DNA probe. PMID- 8236784 TI - In situ hybridizations of Eperythrozoon suis visualized by electron microscopy. AB - Eperythrozoon suis is an extracellular red blood cell parasite that causes icteroanemia and poor growth performance in feeder pigs and has been associated with anemias in baby pigs and reproductive failures in sows. At present, few efficient tests are available for the diagnosis and study of E. suis infection in swine. This report discusses how a recently developed recombinant DNA probe (KSU 2) specific to E. suis DNA was utilized in in situ DNA hybridizations that couple biotinylated TaqI digested products of KSU-2 DNA with an immunogold detection system allowing the visualization by transmission electron microscopy of the gold particles within eperythrozoon organisms. Specific labelling by the probe of eperythrozoon organisms demonstrated a pattern that changed with the various stages of the eperythrozoon life cycle. PMID- 8236785 TI - Bacterial isolates from blood cultures of dogs undergoing dentistry. PMID- 8236786 TI - Effects of exercise and polysulfated glycosaminoglycan on the development of osteoarthritis in equine carpal joints with osteochondral defects. AB - This study assessed the effects of postoperative exercise and intra-articular polysulfated glycosaminoglycan (PSGAG) on the repair of osteochondral defects in the carpal joints of ponies. Eighteen ponies with normal carpi had osteochondral defects (mean dimensions 2.4 cm x 0.9 cm) created arthroscopically on the dorsal aspect of the distal articular surface of the radial carpal bone. The ponies were randomized (while balancing for age [range, 2 to 15 years; median, 5.0 years]) to two groups--nine ponies were exercised and nine were stall confined. Beginning at surgery, six ponies in each group received five weekly intra-articular injections of PSGAG (250 mg) in one joint and lactated Ringer's solution in the contralateral joint; the remaining three ponies in each group received lactated Ringer's solution in both joints. The incremental exercise schedule on a circular, rotating walker was begun six days after surgery and occurred twice daily, reaching a maximum of 0.7 miles of walking and 2.7 miles of trotting by the third postoperative month. The effects of treatment on the joint tissues were determined by weekly lameness examinations and measurement of the range of carpal joint motion, carpal radiographs at six and 17 weeks after surgery, synovial fluid analysis, and cytologic evaluation of alcohol-fixed synovial fluid specimens at weeks 1 through 4 and week 17, and histology of the synovial membrane. Ultrasound images of the carpi were acquired before operation and at weeks 1, 2, 4, 8, 10, 13, and 17. Ponies were euthanatized 17 weeks after surgery. Exercise, without medication, caused more lameness throughout the study compared with no exercise. Exercised, nonmedicated ponies had the greatest limitation to carpal flexion (more painful joints), and nonexercised, nonmedicated (control) ponies had the least limitation to flexion. Radiographic scores indicated that the exercised, nonmedicated ponies had significantly (p < .05) more signs of osteoarthritis than exercised, medicated and control ponies. Ultrasonographic measurements indicated that exercise, without medication, caused the greatest increase in combined measurement of the joint capsule thickness and synovial fluid accumulation at all postoperative times. Synovial lining cell numbers in the synovial fluid from exercised ponies were significantly (p < .05) higher than in nonexercised ponies at week 1, and this trend continued at weeks 4 and 17 (p < .1). There were significantly (p < .05) more morphologic abnormalities in the synovial lining cells from exercised than from nonexercised ponies at week 17. Medication with PSGAG enabled exercised carpal joints to be flexed significantly further from weeks 2 through 6 compared with nonmedicated joints.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8236787 TI - Effects of neodymium:yttrium aluminum garnet laser irradiation on endometrium and on endometrial cysts in six mares. AB - Effects of neodymium:yttrium aluminum garnet (Nd:YAG) laser irradiation on equine endometrium were evaluated in vitro and in six mares with endometrial cysts. The Nd:YAG laser was applied to six endometrial sites, in each of five uterine specimens, with power densities of 5659 to 33,954 J/cm2. Depth of tissue ablation was measured and graded on histologic sections of the tissue lesions. Power density had a significant effect on the depth of tissue ablation (p < .001). Grade 3 lesions (full-thickness ablation of the endometrium) were created with energy densities of 16,977 to 33,954 J/cm2. Six mares had endometrial cysts treated by photoablation. Two of the four mares that were reproductively sound but barren, despite appropriate breeding, produced foals after treatment. One mare remained reproductively unsound after treatment, and another mare that was treated postpartum was bred successfully. PMID- 8236788 TI - Development of parenchymal abdominal organ models for use in teaching veterinary soft tissue surgery. AB - Models of the canine spleen, kidney, and liver were made from soft plastic to simulate the organs of the live animal as closely as possible in appearance and tissue handling properties. Each organ model was independently evaluated by five small animal surgeons who performed several common surgical procedures on each model. All models had a realistic appearance and, with the exception of one tissue handling problem with the kidney model, and one with the liver model, tissue handling properties of the models were comparable to those of the organs in the live animal. All models were useful for teaching each of the procedures evaluated. PMID- 8236789 TI - Transsplenic portal catheterization. Surgical technique and use in two dogs with portosystemic shunts. AB - The portal vasculature can be accessed by using a through-the-needle catheter system to pass a catheter through the splenic parenchyma and into a major splenic vein at the hilus. The authors have termed this technique transsplenic portal catheterization (TPC). Transsplenic portal catheterization is indicated for portal angiography, portal pressure measurement, and chronic portal blood sampling. Clinical applications of this technique include use in diagnosis and surgical management of portosystemic shunts and differentiation of prehepatic, hepatic, and posthepatic hypertension. This report describes the technique of transsplenic portal catheterization. Clinical use of this technique in two cases of portosystemic shunts are included. PMID- 8236790 TI - Electroencephalography of detomidine-ketamine-halothane and detomidine-ketamine isoflurane anesthetized horses during orthopedic surgery. A comparison. AB - This study was done to compare the electroencephalographic (EEG) response evoked by orthopedic surgery in halothane- and isoflurane-anesthetized horses. Eight horses scheduled for bilateral arthroscopic surgery of the stifle were premedicated with detomidine (20 micrograms/kg) intravenously and five minutes later induced to anesthesia with ketamine (2.2 mg/kg) intravenously. Anesthesia was maintained with either halothane or isoflurane. Assignment of inhalation anesthetic was done randomly. The multiple of minimal alveolar concentration (MAC) of halothane required for anesthesia was significantly higher than the multiple of MAC of isoflurane (p < .05) required. Total amplitude of the EEG with halothane was smaller than with isoflurane (p < .05), but 13.0 to 32.0 Hz high frequency/0.0 to 3.9 Hz low frequency (beta/delta) ratio was greater for halothane (p < .05). Arterial partial pressure of oxygen (PaO2) was significantly (p < .05) higher with isoflurane than with halothane. The differences in EEG frequency shift observed suggest that isoflurane provided better analgesia than halothane for this group of horses. PMID- 8236791 TI - Age-related increase of porcine natural interferon alpha producing cell frequency and of interferon yield per cell. AB - Porcine blood mononuclear cells (PBMC) were shown to secrete interferon alpha (IFN-alpha) after induction by a coronavirus, the transmissible gastroenteritis virus (TGEV). IFN-alpha producing cells, referred to as natural interferon alpha producing (NIP) cells, were detected by an ELISPOT assay using anti-porcine IFN alpha monoclonal antibodies. The frequency of NIP cells among blood cells is low, at most 40-110 per 10(5) PBMC and each NIP cell was found to produce several units of IFN. We have shown that NIP cell frequency and IFN yield per cell gradually increased with the age of the donor animals, from the neonatal period to the adult age, with a significant increase around puberty. Our present results also indicate that NIP cells may be influenced by physiological and genetic factors; thus (1) NIP cell frequency and IFN yield per cell were decreased during lactation; (2) Chinese (Meishan) pigs were found to have higher NIP cell frequency and IFN yield per cell than European (Large White) animals. PMID- 8236792 TI - Secretion of co-stimulatory cytokines by monocytes and macrophages during infection with Trypanosoma (Nannomonas) congolense in susceptible and tolerant cattle. AB - Bovine macrophages and monocytes were cultured in vitro and analyzed for their capacity to secrete co-stimulatory cytokines. To this end, the culture medium was titrated on suboptimally stimulated murine thymocytes. A low residual release by normal monocytes was noted which usually remained below the detection limit of the assay. These cells could be induced to secrete high titres following activation with bacterial lipopolysaccharide. When harvested from animals infected with Trypanosoma congolense, the cells released high titres spontaneously. This increase in co-stimulatory cytokine secretion was noted in both peripheral blood monocytes and splenic macrophages and was amplified by addition of indomethacin. The activation was transient, and the titres had dropped to pre-infection values at the end of the experiment. At that time, the monocytes were, however, still able to respond to external stimuli. Addition of neutralizing anti-transforming growth factor beta antibodies did not influence the thymocyte co-stimulatory activity of the supernatants. High levels of co stimulatory cytokine secretion were noted with monocytes from both the susceptible Boran breed and the tolerant N'Dama breed. Early in infection, at Day 10 post infection, the production by the N'Dama monocytes was 16 times higher than the production by the Boran monocytes. Later in the infection, the titres were similar in both breeds. PMID- 8236793 TI - Generation and partial characterization of an eosinophil chemotactic cytokine produced by sensitized equine mononuclear cells stimulated with Strongylus vulgaris antigen. AB - Supernatants generated by stimulation of peripheral blood mononuclear cells (PBMC) from Strongylus vulgaris sensitized or immunized ponies were assayed in vitro for eosinophil chemotactic activity (ECA) using the filter system in blind well chambers. The supernatants from these cultures were chemotactic for eosinophils, but not for neutrophils. Supernates from cultures of unsensitized PBMC stimulated with S. vulgaris antigen were not chemotactic for eosinophils. ECA was first detected in culture supernatants after 1.5 h of incubation and was dependent on both antigen and PBMC concentrations, but independent of serum concentrations. Both female and male S. vulgaris worm antigens stimulated ECA production from sensitized PBMC. ECA was not induced by in vitro stimulation of sensitized S. vulgaris PBMC by female Strongylus edentatus worm antigen. Partial characterization of the eosinophil chemotactic cytokine showed it to be nondialyzable, greater than 8000 molecular weight (MW), and sensitive to heating (56 and 95 degrees C), trypsin, and sodium metaperiodate treatments, suggesting that the cytokine is a protein containing some essential carbohydrate moieties. The cytokine described in this paper could partially contribute to the in vivo blood and tissue eosinophilia in experimental S. vulgaris infection. PMID- 8236794 TI - Newcastle disease virus: virus replication in the harderian gland stimulates lacrimal IgA; the yolk sac provides early lacrimal IgG. AB - An indirect immunoperoxidase monolayer assay was used to measure the location of virus and class-specific antibodies to Newcastle disease virus (NDV) of chickens. The intra-ocular vaccination of chicks with the Ulster and Hitchner B1 strains of NDV resulted in the highest titre of virus being recovered from the Harderian gland (HG) at 10(6) iu/0.1 g tissue without any accompanying virus in the faeces. Maternal IgG antibody which was detected in the lacrimal fluid at 1/50 of serum levels of IgG antibody could prevent this replication. Lacrimal IgA antibody to NDV was shown to be absolutely dependent on local virus replication for its production because it reached titres of 10(3) after local intra-ocular infection compared with < 10(1.5) after two intravenous inoculations of inactivated virus. Biliary IgA, by contrast, reached 10(3.5) after either immunisation schedule. Lacrimal IgG and IgM antibody to NDV occurred at 1-9% of the titres of serum antibody to NDV after immunisation with inactivated virus or the passive transfer of NDV-immune serum between chickens. This percentage increased to 13-33% of serum titres after intra-ocular infection with NDV as if the local replication of NDV in the HG stimulated lacrimal antibody of all classes. This is the first report of NDV replication in the HG and of virus-specific IgM in the lacrimal fluid. PMID- 8236795 TI - Local antibody forming cell responses to the Hitchner B1 and Ulster strains of Newcastle disease virus. AB - The Hitchner B1 and Ulster strains of Newcastle disease virus (NDV) replicated to high titre in the Harderian gland (HG) after eye-drop infection. The Harderian gland then became the major site of antiviral IgA-antibody-forming cells (AFC) in the body and their number correlated to the level of antiviral IgA antibody in the tears. The spleen, HG and femoral bone marrow all contained comparable levels of antiviral IgG-AFC and IgM-AFC after two intra-ocular inoculations of virus, whereas the caecal tonsil and bursa contained few AFC despite the local replication of the Ulster strain of NDV leading to high titres of virus in the faeces. Vaccines of the Hitchner B1 strain of NDV were much less effective at inducing antibody by the intranasal compared with intra-ocular route and no virus was re-isolated after intranasal vaccination. The intravenous inoculation of inactivated Iscoms of NDV could stimulate the spleen, but not the Harderian gland to the same extent as a live virus. PMID- 8236796 TI - Isolation of porcine IgE, and preparation of polyclonal antisera. AB - High Titre, Ascaris specific, porcine reaginic antibody was induced by combining parasitic infestation and antigen challenge. The reagin was characterised as being heat labile at 56 degrees C, possessing long term skin sensitising activity, and as having physicochemical properties similar to IgE in other species. Porcine IgE was purified by conventional immunochemical techniques, and an epsilon chain isolate prepared using SDS-PAGE gel excision and electro elution. Antisera raised against the intact IgE molecule and its epsilon chain were rendered monospecific by immunoabsorption and characterised by reverse cutaneous anaphylaxis (RCA) testing, passive cutaneous anaphylaxis inhibition, immunohistology and Western blotting. These antisera are currently being used to study the regulation of IgE responses in the pig. PMID- 8236797 TI - Effects of certain inflammatory mediators on bovine neutrophil migration in vivo and in vitro. AB - Migration of bovine neutrophils towards endotoxin, recombinant bovine interleukin 1 beta (rBoIL-1 beta), recombinant human tumor necrosis factor-alpha (rhTNF alpha), platelet-activating factor (PAF), complement factor C5a, leukotriene B4 (LTB4), and recombinant human interleukin-8 (rhIL-8) was studied in vivo, using the teat cistern model, and in vitro using the modified Boyden chamber method. Infusion of endotoxin, rBoIL-1 beta, rhTNF-alpha, PAF, or C5a into the teat cistern induced significant accumulation of leukocytes, mainly neutrophils, during the sampling period. Endotoxin was, on a molar basis, the most potent inducer of cell accumulation in vivo, followed by rBoIL-1 beta, while C5a, PAF and rhTNF-alpha were less potent. No significant cell accumulation was observed after infusion of LTB4 or rhIL-8. A significant migration of cells into the teat cistern was first observed 2 h after the infusion of endotoxin or rBoIL-1 beta, the rBoIL-1 beta-induced response started somewhat earlier. The first significant cell accumulation after infusion of PAF or C5a was observed already 1.5 h post infusion. The largest numbers of cells were reached 2.5-4.5 h after the infusion of endotoxin, rBoIL-1 beta, rhTNF-alpha, PAF or C5a. In vitro, significant migration of bovine blood neutrophils was observed towards C5a or rhIL-8, and to a lower extent towards LTB4, while no chemotactic response to endotoxin, rBoIL-1 beta, rhTNF-alpha, and PAF was observed. Possible roles of the different substances as inducers of neutrophil migration into the bovine teat are discussed. PMID- 8236798 TI - Production and characterization of mouse monoclonal antibodies directed against canine IgE and IgG. AB - Immediate-type hypersensitivity reactions have been studied infrequently in dogs, in part because of limited availability of antisera specific for canine IgE. A series of murine hybridoma cell lines were prepared, that produced monoclonal antibodies (MAb) with specificity for canine immunoglobulin E (IgE) and IgG. The MAb were tested for their ability to induce a reverse cutaneous anaphylaxis reaction in dog skin, to neutralize the Prausnitz-Kustner reactivity of atopic dog serum, to serve as a ligand in immunoaffinity chromatography, and to bind to IgE and other Ig subclasses in several ELISA systems. Some of the MAb produced were found to be specific for canine IgE. Other MAb recognized common or similar determinants on IgE and IgG, or on IgG and IgM, though with apparently differing affinities. Heat or acid treatment of canine IgE abolished most, but not all, of the reactivity with the anti-IgE MAb. These MAb will be useful for further study of IgE-mediated phenomena in the dog. PMID- 8236799 TI - Mitogen and antigen induced B and T cell responses of peripheral blood mononuclear cells from the harbour seal (Phoca vitulina). AB - In vitro assays were developed for studies concerning the functioning of the immune system of the harbour seal (Phoca vitulina). Proliferative responses of peripheral blood mononuclear cells (PBMC) were measured after stimulation with different concentrations of the mitogens concanavalin A (Con A), pokeweed mitogen (PWM), phytohaemagglutinin (PHA) or lipopolysaccharide from Salmonella typhimurium (LPS). Con A and PWM induced strong proliferative responses, while PHA and LPS induced comparatively low proliferative responses. Responses of mitogen stimulated PBMC to recombinant human interleukin-2 (rhIL-2) and in vitro immunoglobulin production by mitogen stimulated PBMC were measured to discriminate between stimulation of T cells and B cells. It was found that Con A and PHA stimulate phocine T cells, PWM stimulates both T cells and B cells and LPS predominantly stimulates phocine B cells. Antigen-specific immune responses were measured after immunization of seals with an inactivated rabies vaccine and/or with tetanus toxoid. Antigen-specific proliferation of PBMC and the presence of antigen-specific antibody forming cells were demonstrated for both antigens in the PBMC of immunized animals. The responses measured in vitro correlated well with the development of specific serum antibody titers to these antigens. PMID- 8236800 TI - Isolation of the porcine complement activation fragments C3c and C3d and their use in the preparation of monospecific antibodies. AB - The hemolytic test to date has been the sole analytic technique applied to study the complement reaction in the swine. To improve the analytical possibilities for this species we have developed polyclonal antibody reagents with specificities for the C3c and C3d activation fragments of swine C3. Access to these reagents, by which activation products can be analysed in tissues and biological fluids, will offer new possibilities for a more precise analysis of the complement reaction. PMID- 8236801 TI - Phagocytosis, killing, and oxidant production by bovine monocyte-derived macrophages upon exposure to Brucella abortus strain 2308. AB - Phagocytosis and intracellular survival of Brucella abortus, and oxidant production by monocyte-derived macrophages from ten B. abortus-naive cows were studied. Phagocytosis of bacteria opsonized with naive-autologous sera or reactor serum was significantly less than phagocytosis of bacteria opsonized with fetal bovine serum. After phagocytosis, intracellular survival of bacteria opsonized with naive-autologous or reactor sera was significantly less than survival of bacteria opsonized with fetal bovine serum. Production of oxidant by macrophages stimulated with B. abortus opsonized with naive-autologous, reactor, or fetal bovine sera was not significantly different. Although macrophages from one animal showed significantly less phagocytic activity, intracellular killing and oxidant production by macrophages from the ten individual cows toward B. abortus opsonized with naive-autologous, reactor, and fetal calf sera were homogeneous. The abilities of the macrophages to phagocytize and to kill B. abortus were not associated with each other or with oxidant production. Innate resistance or sensitivity to B. abortus was not identified in the cows based on macrophage function. PMID- 8236802 TI - Evaluation of whole cell and subcellular vaccines against Brucella ovis in rams. AB - Five antigen preparations from Brucella ovis strain REO 198 were incorporated with the pluronic polymer L-121 and muramyl dipeptide and tested as vaccines against B. ovis infection of rams. The antigenic preparations were: (1) a fraction enriched in outer membrane proteins and rough lipopolysaccharide (hot saline extract, HS); (2) the proteins from HS substantially free of lipopolysaccharide; (3) outer membrane blebs; (4) outer membrane-peptidoglycan complexes extracted with detergent; (5) killed whole cells. The experimental vaccines were compared with two standard vaccines, rough Brucella abortus 45/20 whole killed cells in an oil based adjuvant, and live Brucella melitensis Rev 1. Immunizations with non-living vaccines were performed on two occasions, 18 weeks apart. The rams were challenged with a virulent strain of B. ovis 31 weeks after the second vaccination and slaughtered 15 weeks thereafter. Rates of infection in groups vaccinated with Rev 1 (33%), and HS (40%) were significantly lower (P < 0.005 and P < 0.025, respectively) than that in the non-vaccinated control group (87%). Strain 45/20 was the only other vaccine that conferred a significant level of protection (50%) (P < 0.05). The organ distribution of the infection and the level of colonization of infected organs did not differ significantly between infected animals in the various vaccine groups and those in the unvaccinated control group. No statistically significant relationship was detected between the magnitude of the antibody responses to the HS extract, to outer membrane proteins, or to the rough lipopolysaccharide, and freedom from infection. The results indicate that the HS extract of B. ovis may represent a useful alternative to B. melitensis Rev 1 or B. abortus 45/20 as a vaccine against B. ovis. PMID- 8236803 TI - Role of Harderian gland on differentiation and proliferation of immunoglobulin A bearing lymphocytes in chickens. AB - The mechanism of accumulation of surface immunoglobulin A (sIgA) bearing cells in chicken Harderian gland (HG) was examined. Almost no sIgA-bearing cells were identified in HG of 1.5-week-old chickens. In 3.5-week-old chickens, however, 46.4% of the HG lymphocytes were sIgA-bearing cells. When bursa of Fabricius (BF) lymphocytes were transferred into recipients, BF lymphocytes migrated into the HG but the number of these lymphocytes was extremely small. In addition, no correlation was found between the surface immunoglobulin class of BF lymphocytes that migrated into HG and the migration. HG lymphocytes were co-cultured with HG whole cells (which contained 50-60% configurationally epithelial cells) in a non contacting manner for 80 h using an Intercell culture vessel. The results showed a significant increase in the ratio of sIgA-bearing cells in HG lymphocytes compared with the control which was cultured without placing HG whole cells in the external well. B lymphocytes that migrated into HG seemed to undergo specific differentiation in situ into sIgA-bearing cells owing to microenvironmental factors, probably humoral factors excreted from HG epithelial cells, and subsequently undergo proliferation to result in accumulation of sIgA-bearing cells in HG tissue. PMID- 8236804 TI - Analysis of macrophage functions in Rous sarcoma-induced tumor regressor and progressor 6.B congenic chickens. AB - Macrophage functional competence was studied in two congenic chicken lines 6.6-2 (B2B2) and 6.15-5 (B5B5) which are regressors and progressors, respectively, of Rous sarcoma-induced tumors. Sephadex-elicited abdominal exudate cells (AEC) were harvested from 4-week-old chickens to determine their total number, glass adherence potential, percentage of adherent macrophages and phagocytosis of antibody coated (ops) and uncoated (unops) sheep red blood cells (SRBC). Tumoricidal abilities of culture medium conditioned with lipopolysaccharide treated macrophages and of macrophages cocultured with target cells were assessed against 51Cr-labelled tumor cell targets. The congenic lines did not differ in total AEC or percent macrophages. However, AEC from B5B5 birds exhibited significantly lower (P < 0.05) glass-adherence potential than AEC from B2B2 birds exhibited significantly lower (P < 0.05) glass-adherence potential than AEC from B2B2 birds. The percentage of phagocytic macrophages did not differ between lines for unop-SRBC, whereas a higher percentage of B5B5 compared with B2B2 birds (P < 0.05) macrophages phagocytized ops-SRBC. Macrophages from B5B5 birds had significantly (P < 0.05) lower activity in both tumoricidal tests. These results imply that the tumor progression in B5B5 birds is associated with reduced activation of macrophages towards a tumoricidal pathway. PMID- 8236805 TI - Estimation of the genetic variation in complement activity of common carp (Cyprinus carpio L.). AB - The complement status of hybrid, laboratory raised carp was determined by an in vitro approach of the alternate complement activity (ACH50) and total haemolytic activity (CH50), and by measurement of serum C3 levels. The lysis of target sheep red blood cells (RBC) in the haemolytic assay for CH50 activity depended, amongst others, on the haemolysin concentration in the assay. Rocket electrophoresis showed a mean serum C3 concentration of 0.95 mg ml-1. The variation for both ACH50 and CH50 haemolytic activity was approximately 30%. The degree of genetic determination of the parameters was investigated by estimation of their repeatabilities, which were relatively high for CH50 (0.71) and ACH50 activity (0.72), but lower for C3 levels (0.54). Correlations between ACH50 values and C3 levels were significant, but moderate (0.54-0.58). PMID- 8236806 TI - Effect of bovine serum albumin on passive transfer of immunoglobulin G1 to newborn calves. AB - The molecular mechanism in the intestine of newborn calves that results in transfer of intact colostral immunoglobulin from the lumen to the circulation also is capable of transferring a variety of non-immunoglobulin macromolecules. If the capacity of this mechanism is limited, transfer of a large amount of non immunoglobulin protein may interfere with transfer of immunoglobulin. In this experiment, efficiency of IgG1 transfer in newborn calves was reduced from 59 to 36% by the addition of bovine serum albumin (37 mg ml-1) to colostral whey, while the addition of a similar mass of amino acids in the form of acid hydrolyzed casein (37 mg ml-1) did not detectably alter IgG1 transfer. Reduced IgG1 absorption efficiency in calves fed colostrum with added bovine serum albumin is consistent with a limited capacity for the macromolecular transport mechanism in the intestine of newborn calves. PMID- 8236807 TI - Polymorphonuclear neutrophil granulocyte chemotactic hyperresponsiveness in a case of canine acromegaly. AB - Growth hormone (GH) has recently been shown to affect polymorphonuclear neutrophil granulocyte (PMN) function and to be secreted by mononuclear cells, indicating that the hormone may be active in an immunophysiologic network, acting as an endo- or paracrine priming agent. The purpose of the present study was to evaluate the chemotactic responsiveness of canine peripheral PMN in a dog with acromegaly, caused by spontaneous, progesterone-induced hypersecretion of GH and, secondary to this, a seven-fold increase in insulin-like growth factor I (IGF-I). The chemotactic responsiveness towards zymosan-activated serum (ZAS) and leukotriene B4 (LTB4) was evaluated at a time when the dog suffered from acromegaly and again 57 days after corrective surgery (ovariohysterectomy). The experiments showed that PMN from the patient exhibited enhanced chemotactic migration that appeared to be associated with the hypersomatotropic condition as judged from the reversibility of the phenomenon. The glucose intolerance and elevated serum alkaline phosphatase that were observed in the acromegalic dog were also shown to be reversible following surgery. PMID- 8236808 TI - Effect of low temperature storage on the haemolytic complement of cattle and sheep. AB - Sera from cattle and sheep stored at -70, -20 and -10 degrees C were examined for haemolytic complement at Days 0, 37, 54, 86, 133 and 245. The decline in complement followed a similar trend, both by classical pathway (CPW) and alternative pathway (APW) assay methods. Both cattle and sheep sera retained 80% of their complement activity till Day 245 at -70 degrees C whereas at -20 degrees C cattle sera had lost 67% and sheep sera 80% of the activity. At -10 degrees C the decline in complement levels was, in general, similar to -20 degrees C. The complement level of sheep sera declined more rapidly than that of cattle. For reliable results, it is suggested that sera of cattle and sheep should be assayed for complement within 4-5 months at -70 degrees C and within a month at -20 or 10 degrees C. PMID- 8236809 TI - Opsonization of Anaplasma marginale mediated by bovine antibody against surface protein MSP-1. AB - Antibody from cattle immunized with purified major surface protein-1 (MSP-1) was demonstrated to significantly enhance phagocytosis of Florida strain Anaplasma marginale by bovine macrophages in vitro. Serum immunoglobulin from individual MSP-1 immunized, protected cattle varied in ability to promote phagocytosis, however all sera were significantly opsonic as compared with sera from sham immunized control cattle. PMID- 8236810 TI - Diffuse axonal injury: windows for therapeutic intervention allowed by its pathobiology. PMID- 8236811 TI - Immunohistochemical study of copper-zinc and manganese superoxide dismutases in the lungs of human fetuses and newborn infants: developmental profile and alterations in hyaline membrane disease and bronchopulmonary dysplasia. AB - To determine the late gestational development of copper-zinc (CnZn) and manganese (Mn) superoxide dismutases (SOD) in human lung, immunohistochemical localization was performed for each SOD. The lung samples were taken from five aborted fetuses, four fetuses in which intrauterine death occurred, one full-term neonate, two premature infants with hyaline membrane disease and one premature infant with bronchopulmonary dysplasia (BPD). Morphometry was performed, and the percent area of positive staining was computed. The bronchial epithelium was intensely stained from the early stages of gestation (i.e. 17 weeks), while the staining intensity for both CuZnSOD and MnSOD in the peripheral airways increased gradually during lung development. The mean percent area of the staining for CuZn SOD and MnSOD from 16 to 38 weeks was increased 30-fold and 8-fold, respectively, and further increases were observed postnatally. CuZnSOD staining was markedly decreased in lungs with respiratory disorders. However, proliferating type II pneumocytes were intensely stained for MnSOD in the BPD lungs, making the staining area 3-fold larger than that in the control lungs. These results clearly depict age-related increases in staining for both CuZnSOD and MnSOD and an alteration in SOD distribution associated with neonatal respiratory disorders. PMID- 8236812 TI - Abnormal distribution of cathepsin proteinases and endogenous inhibitors (cystatins) in the hippocampus of patients with Alzheimer's disease, parkinsonism dementia complex on Guam, and senile dementia and in the aged. AB - The immunolocalization of cathepsins B(CB), H and L and cystatins alpha(C alpha) and beta(C beta) were examined in the hippocampus of cases of sporadic Alzheimer's disease (12 cases), parkinsonism-dementia complex on Guam (eight cases), senile dementia of Alzheimer type (two cases), aged subjects with marked senile change (one case) and controls (12 cases, including six normal subjects). CB was lower in most nerve cells in patients than in controls, but markedly increased at the sites of intracellular neurofibrillary tangles (NFTs) and degenerative neurites and/or dendrites in and outside senile plaques (SPs), indicating its close involvement in the metabolisms of various proteins in NFT and SPs. Abundant C alpha and C beta were demonstrated in SP amyloid, suggesting that they are amyloid constituents or co-exist with amyloid. The present study indicated that CB, C alpha and C beta are closely involved in abnormal protein metabolism in NFTs and SP amyloid and suggested that degeneration or denaturation of intracellular proteins, including substrates for proteases and lysosomes, from some acquired cause, results in absolute and/or relative overload for these proteolytic systems, including their inhibitors. This results in incomplete and/or abnormal proteolysis related to NFT and/or amyloid formation. PMID- 8236813 TI - Prognostic significance of spontaneous tumour necrosis in osteosarcoma. AB - Preoperative chemotherapy is an integral part of the management of osteosarcoma, and the extent of tumour necrosis found at operation is an important prognostic variable. Knowledge about spontaneous, pretherapy necrosis is difficult to obtain but provides important quantitative information about the necrotic effect of chemotherapy. Using three different methods, we studied spontaneous tumour necrosis in 20 localized intramedullary osteosarcomas of the distal femur diagnosed between 1963 and 1972. All patients received surgical treatment only. All six patients with spontaneous necrosis involving more than 20% of tumour died. Five of 14 patients with necrosis amounting to less than 20% were long term, disease-free survivors. The extent of necrosis was independent of tumour size. Two semiquantitative methods of evaluation were easily applied and reproducible. Spontaneous necrosis in untreated osteosarcomas occurs frequently; extensive necrosis may indicate a rapid clinical course. Tumour necrosis can be quantified reliably in clinical work. PMID- 8236814 TI - Three-dimensional analysis of increased vasculature around the glomerular vascular pole in diabetic nephropathy. AB - In diabetic nephropathy, several small vessels are frequently observed around the glomerular vascular pole in addition to the usual afferent and efferent arterioles. To elucidate the morphology of these abnormal small vessels, a three dimensional study was performed by using computer-aided reconstruction techniques. In the present study, the renal tissue samples of 21 biopsy and 73 autopsy cases of diabetic glomerulonephropathy were examined. In addition to ordinary light microscopic observations, three series of serial sections from one autopsy and two biopsy cases were analysed. Five glomeruli with increased numbers of vessels around the vascular pole were reconstructed three-dimensionally. The vasculature in and around the glomerulus was analysed in detail by rotating and viewing in different planes via computer-generated three-dimensional images. These vessels anastomose to the lobular structure of the intraglomerular capillary network, mainly to the afferent branches through the widened vascular hilus. The distal end of the vessels anastomoses to the peritubular capillary. The increased vasculature is interpreted as neoangiogenesis resulting from diabetes, which may have a functional role in facilitating efferent blood flow from the glomerulus. PMID- 8236815 TI - Calcium pump expression in human bone and soft tissue tumours. AB - Ninety-one cases of human bone and soft tissue tumours were studied for calcium pump expression by strepto-avidin-biotin immunohistochemical staining with a monoclonal antibody against sarcoplasmic reticulum calcium-ATPase (mAb6F5). Two out of 5 cases of embryonal rhabdomyosarcoma, 1 out of 5 cases of biphasic synovial sarcoma, 4 of 4 cases of chordoma and all of 3 chondrosarcoma cases were positive for mAb6F5. Although this novel monoclonal antibody can be used as a marker of myogenic tumours, the present positive result for endoplasmic reticulum calcium-ATPase (calcium pump) in other tumours including chordoma, chondrosarcoma and synovial sarcoma indicates a wider immunoreactivity. The findings further suggest that intracellular calcium may play an important role in cell proliferation and/or differentiation. PMID- 8236816 TI - Localization of metallothionein in breast carcinomas. An immunohistochemical study. AB - Metallothionein (MT) is a cysteine-rich, low molecular weight protein that binds zinc, copper, and cadmium. It is present in a number of normal cells including hepatocytes particularly during fetal and early postnatal life. It has been suggested that developmental profile of MT is similar to other oncofetal gene products and hence, it could be used as a marker for aggressive tumour behaviour. In order to test that hypothesis, we used a monoclonal antibody to MT and immunohistochemically evaluated formalin-fixed, paraffin-embedded tissues from 79 breast carcinomas. In non-neoplastic breast tissue, a strong nuclear and cytoplasmic staining was observed in myoepithelial cells. Positive staining for MT was present in 35 (44%) of breast carcinomas. In most positive cases, nuclear, or both nuclear and cytoplasmic staining was seen. All positive tumours were invasive ductal carcinomas, including a medullary and a metaplastic carcinoma. None of the mucinous, lobular, or intraductal papillary carcinomas reacted for MT. A statistically significant association was found between MT immunostaining and histological grade (P < 0.01) as well as with nuclear grade (P < 0.01). We also observed an inverse relationship between MT staining and oestrogen receptor content of tumours (P < 0.01). Similarly, a statistically significant association was found between moderate and strong MT immunostaining and decreased overall survival and shorter disease-free survival (P < 0.01). MT immunostaining was also predictive of a worse prognosis in the subgroup of lymph node negative (P < 0.001) and oestrogen receptor negative patients (P < 0.01). No statistically significant association was found between MT staining and size of tumour or the presence of lymph node metastasis. We conclude that MT staining may be a useful marker of less differentiated and more aggressive carcinomas of the breast. PMID- 8236817 TI - Intratesticular serous cystadenoma of borderline malignancy. A pathological, histochemical and DNA content study of a case with long-term follow-up. AB - A 61-year-old man presented with a painless right testicular swelling of 6 months duration. A right orchiectomy was performed and pathological examination showed an intratesticular serous borderline tumour (SBT). Immunohistochemical staining was positive for carcinoembryonic antigen, LeuM1, B72.3, S100-protein, Ca125, cytokeratins AE1/AE3 and vimentin, suggesting a Mullerian origin or differentiation. DNA image analysis revealed an aneuploid histogram. The favorable outcome of the patient confirms that testicular SBTs behave as non aggressive tumours, even when characterized by aneuploid DNA content. PMID- 8236818 TI - Pearson's marrow/pancreas syndrome: a histological and genetic study. AB - A patient with features of Pearson's syndrome who presented with transfusion dependent severe macrocytic anaemia, neutropenia, thrombocytopenia, and insulin dependent diabetes mellitus in the neonatal period is described. His bone marrow was characterized by marked vacuolization of myeloid precursors and ringed sideroblasts. Autopsy examination revealed fibrosis and steatosis of the liver, reduction in the size and number of the islets, fibrosis and acinar atrophy of the pancreas, vacuolation of renal tubules, glomerulosclerosis, and "ragged red" fibres of skeletal muscles. Analysis of mitochondrial DNA (mtDNA) from the autopsied liver and skeletal muscle showed mtDNA heteroplasmy in both tissues, with one population of mtDNA deleted by 7374 bp. The deleted region was bridged by a single nucleotide, C, in normal mtDNA. PMID- 8236819 TI - Familial occurrence of periapical cemental dysplasia. AB - A family with periapical cemental dysplasia is reported. The affected individuals displayed classical features of periapical cemental dysplasia on radiographic examination. The lesions consisted chiefly of radiolucent areas; however, some had central areas of radiodensity. Histopathological examination of one of the lesions revealed fibrous elements containing fused dense sclerotic cemental masses. Familial incidence of florid cemento-osseous dysplasia with an autosomal mode of inheritance has been reported previously. The condition described in this report appears to be different. However, the two conditions may be part of a spectrum occurring in a single genetic entity with the diversity possibly resulting from variable expressivity of a single gene. PMID- 8236820 TI - Perlecan: the multidomain heparan sulphate proteoglycan of basement membrane and extracellular matrix. PMID- 8236821 TI - Verruciform xanthoma of the oral mucosa: a clinicopathological study with immunohistochemical findings relating to pathogenesis. AB - Verruciform xanthoma is an uncommon benign lesion with unknown aetiology and pathogenesis. In this study, we report ten cases of verruciform xanthoma and document their clinical and histopathological findings. An immunohistochemical investigation was performed using antibodies to macrophage, leukocyte common antigen, T lymphocytes, B lymphocytes, S-100 protein, lysozyme and alpha-1 antichymotrypsin. Our results were similar to the other reported cases. Eighty percent of our cases were found on the gingiva. Candidal hyphae were found in the superficial parakeratotic layers in five cases. The clinical diagnosis of the lesion ranged between papilloma and squamous cell carcinoma. It is important for clinicians to take into consideration the possibility of verruciform xanthoma in the differential diagnosis of papillary and granular lesions of oral mucosa. Immunohistochemically, all foam cells were strongly stained with antimacrophage antibodies. T lymphocytes were the predominant infiltrating lymphocytes in the lesion. Langerhans cells in the epithelia were fewer than those in corresponding normal tissue. Our immunohistochemical findings suggest that verruciform xanthoma is may be a local immunological disorder, with a cell mediated mechanism. PMID- 8236822 TI - Cell types infected in human cytomegalovirus placentitis identified by immunohistochemical double staining. AB - Chronic villitis is almost always present in intrauterine infection with human cytomegalovirus (HCMV). The inflammatory response to this virus has been described in detail. However, little is known about the types of placental cells that may be infected by HCMV and six cases of HCMV placentitis were thus investigated to identify the vulnerable cell types. Immunohistochemical double staining analyses were performed using antibodies to HCMV immediate early antigens and to specific cellular marker proteins. Fixed connective tissue cells could be demonstrated to be the predominantly infected cell type in each placental tissue. Endothelial cells and macrophages were also found to be infected in all six cases, whereas evidence of trophoblast infection was obtained in four cases. It is concluded that release of infectious virus by connective tissue cells, macrophages and endothelial cells may play a critical role in transplacental transmission of HCMV. The findings further suggest that the cytopathic effect of HCMV infection on these cells might be involved in the pathogenesis of intrauterine HCMV disease. PMID- 8236823 TI - Immunohistochemical analysis of p53 protein over-expression in endometrial carcinomas: inverse correlation with sex steroid receptor status. AB - Mutations of the tumour suppressor p53 gene have been reported in a variety of human malignant tumours, and are frequently associated with over-expression of p53 protein. To examine the significance of p53 gene alteration in endometrial carcinomas, we studied the immunohistochemical reactivity with a monoclonal antibody against p53 (PAb 1801) in 30 endometrial carcinomas as well as in 64 normal endometria. The presence or absence of correlation of p53 over-expression with the clinicopathological features and with the immunohistochemical expression of sex steroid receptors (oestrogen receptors; ER, progesterone receptors; PR) was also analysed. Expression of p53 was found in none of 64 normal endometria, but was identified in 5 of the 30 (16.7%) endometrial carcinomas. All 5 of the p53-positive tumours developed in women more than 3 years post-menopause, whereas the carcinomas in 5 pre-menopausal women and 3 women less than 3 years post menopause were p53-negative. None of the 5 p53-positive carcinomas was associated with adjacent endometrial hyperplasia. Two of the 5 p53-positive tumours showed non-endometrioid histology: serous papillary and clear cell carcinomas. In contrast, 6 carcinomas accompanied by adjacent hyperplasia were p53-negative. In addition, ER and/or PR expression was found in none of the 5 p53-positive tumours, but was present in 21 of the 25 p53-negative tumours (p < 0.01). These clinicopathological features of p53-positive carcinomas and the inverse correlation of p53 immunoreactivity with sex steroid receptor status suggest that p53 over-expression is frequent in a specific category of endometrial carcinoma, presumably oestrogen-unrelated tumours. PMID- 8236824 TI - Nasopharyngeal angiofibroma: an immunohistochemical study of 32 cases. AB - Thirty-two cases of nasopharyngeal angiofibroma, including 2 recurrences, all of which had been excised from males between 7 and 25 years, were subjected to systematic immunohistochemical study. Most of the tumour vessels, which lacked elastic laminae, were characterized by vascular walls of irregular thickness and variable muscle content. In places endothelial cells were only separated from the stroma by a single attenuated layer of contractile cells, whereas elsewhere the same vessel walls showed pad-like thickenings of their muscle coat. All cells of the vessel walls showed immunoreactivity for vimentin and smooth muscle actin, whereas desmin-positive cells were present only in small numbers in some vessels, generally those with thicker muscle coats. The stromal cells were decorated by vimentin antibodies only; however, in some more fibrotic hyaline areas the stromal cells displayed also reactivity for smooth muscle actin. In most cases S 100 protein-staining disclosed many nerves, and this accentuated their parital distortion by tumour tissue. Our findings provide an extended insight to the morphology of angiofibromas at this site, particularly highlighting the irregularity of their vascular walls, which, taken together with the lack of elastic laminae and elastic stromal fibres, can be held responsible for the typical pronounced tendency for haemorrhage in these lesions. PMID- 8236825 TI - Epidermal growth factor receptor expression in squamous cell lung carcinomas: an immunohistochemical and gene analysis in formalin-fixed, paraffin-embedded material. AB - Epidermal growth factor (EGF) and its receptor (EGFr) constitute an important and well-characterized mitogenic system in various ectodermal tissues. We evaluated the expression of EGFr and examined possible EGFr gene alterations in 18 formalin fixed, paraffin-embedded squamous cell lung carcinomas (SCLC) by an immunohistochemical assay, Southern blotting and differential polymerase chain reaction (DPCR). The immunohistochemical study employing the F4 and EGF-R1 monoclonal antibodies, directed against the intra- and extra-cellular portion of the receptor respectively, showed EGFr over-expression in 89% of the SCLC cases examined. All cases showed positive immunostaining for both antibodies, thus excluding the possibility of truncated receptors. In addition, analysis of the EGFr gene was carried out by Southern blotting and DPCR on paraffin extracted DNA from the same carcinoma cases. We found amplification of the EGFr gene in 5/18 (27%) SCLCs. All 5 positive cases showed EGFr over-expression, suggesting a possible correlation between the presence of EGFr gene amplification and over expression of receptor protein. No correlation was observed among EGFr staining, EGFr gene amplification and differentiation of carcinomas. In addition, Southern blot analysis with HER-A2, a probe which hybridizes a sequence of the receptor's intracellular domain, revealed three novel EcoRI restriction fragment patterns. We suggest that these patterns correspond to EcoRI polymorphic sites of the receptor's tyrosine kinase domain. PMID- 8236826 TI - Fatal infantile hypertrophic cardiomyopathy secondary to deficiency of heart specific phosphorylase b kinase. AB - We describe here a male infant with a rare form of glycogenosis caused by deficiency of heart specific phosphorylase b kinase. The disease phenotype was characterized by severe glycogenosis restricted to the heart muscle with secondary rapidly progressive hypertrophic cardiomyopathy causing death at the age of 47 days. PMID- 8236827 TI - Epithelioid angiosarcoma of the intestinal tract with endothelin-1-like immunoreactivity. AB - Two rare cases of intestinal epithelioid angiosarcoma arising in the sigmoid colon and small intestine are reported. The small intestinal tumours were located in the duodenum and upper jejunum. All tumours showed reddish-black protuberant masses with comparatively clear margins. Histology reveals solid and epithelioid growth of large polygonal cells, mimicking undifferentiated carcinoma. Vascular differentiation such as lamina containing red cells and intracytoplasmic vacuolization is noticed. Intense immunoreactivity to Ulex europaeus agglutinin I and JC70 (CD31), and sporadic positive reaction to factor VIII related antigen were detected in one case but not in the other. However, both tumours have cells which show intense endothelin-1 immunoreactivity. Positive immunostaining occurred with low molecular weight keratins suggesting epithelial differentiation and suggested epithelioid angiosarcoma as a diagnosis, a specific entity within angiosarcoma. Because of their histological features, epithelioid angiosarcomas may be confused with undifferentiated carcinomas. Endothelin-1-like immunoreactivity seems to be a good marker for this type of angiosarcoma, even when the tumour cells lack usual marker substances for endothelial cells. PMID- 8236828 TI - Synovial metaplasia of the skin. AB - We present two female patients, aged 15 and 45 years, with synovial metaplasia of the skin. Both lesions, localized in the knee and hand respectively, arose after preceding local trauma. Case 1 had intradermal irregular cystic spaces in the adjacent myxoid stroma of which large polygonal eosinophilic cells were found. In contrast, case 2 was characterized by a longitudinal space within the lower dermis and subcutis which was lined by a membrane similar to hyperplastic synovium. The cells of the membrane showed an eosinophilic spindle shaped cytoplasm with processes towards the lumen. In both cases the eosinophilic cells, strongly suggestive of fibroblasts, showed staining for vimentin only, whereas no reactivity could be obtained with antibodies to actin, desmin, S-100 protein, Factor VIII related antigen, cytokeratin, epithelial membrane antigen, carcinoembryonic antigen and CD 68. The absence of CD 68 positivity differs from that seen in normal and hyperplastic synovium. Delayed wound healing around a nidus seems the most likely cause of the change. PMID- 8236829 TI - Multiple papillary adenomas of type II pneumocytes found in a 13-year-old boy with von Recklinghausen's disease. AB - A case of multiple papillary adenomas of type II pneumocytes is reported. A 13 year-old boy with von Recklinghausen's disease had small nodular lesions in both lungs without symptoms. The biopsied lung contained greyish-white nodules ranging in size from 0.5 to 2 mm. Light microscopic examination revealed cuboidal to low columnar cells arranged in a papillary pattern. Elastic fibres were present in the tumour stroma. Electron microscopically, the cells had osmiophilic lamellar bodies in the cytoplasm and short microvilli along the free border. The tumour cells expressed immunoreactivity for epithelial membrane antigen and surfactant apoprotein antibodies. More than 6 years after open lung biopsy, the patient is well but small nodular shadows can still be identified. PMID- 8236830 TI - [Eradication of smallpox]. PMID- 8236831 TI - [HIV-1/AIDS epidemic in Southeast Asia]. PMID- 8236832 TI - [Rinderpest eradication and recombinant vaccine]. PMID- 8236833 TI - [Viral superantigens]. PMID- 8236834 TI - [Mechanism of viral antigen processing and presentation]. PMID- 8236835 TI - [Role of laboratory network in polio eradication]. PMID- 8236836 TI - [Cytotoxic T cell response to viral infection]. PMID- 8236837 TI - [Vaccine and protection against infection]. PMID- 8236838 TI - [Current status of information exchange program between Chinese and Japanese virologists]. PMID- 8236839 TI - [Present status and prevention of measles in developing countries]. PMID- 8236840 TI - [Present status of acute respiratory infections in developing countries and the countermeasures]. PMID- 8236841 TI - [Present status of viral diarrhea in developing countries and the countermeasures]. PMID- 8236842 TI - [Present status of rabies in developing countries and the countermeasures]. PMID- 8236843 TI - [Present status of viral haemorrhagic fevers in developing countries and the countermeasures]. PMID- 8236844 TI - [Flavivirus infections in tropical areas]. PMID- 8236845 TI - [Viral hepatitis--control of hepatitis B in Asia]. PMID- 8236846 TI - [Serological survey of cytomegalovirus infections in the north-east of Brazil]. PMID- 8236847 TI - [Present status of HTLV-I infections in developing countries and the countermeasures]. PMID- 8236848 TI - Chromatic aberration and accommodation: their role in emmetropization in the chick. AB - The roles of chromatic aberration and accommodation as cues to emmetropization in the chick were investigated. Myopia was induced monocularly by lid suture for a period of 1-2 weeks from hatching, after which eyes were reopened and the recovery process followed. Monochromatic light (ML) rearing conditions and ciliary nerve section surgery were used to eliminate chromatic aberration and accommodative activity respectively. Control animals were reared in white light (WL). When accommodation was left intact, chickens reared under monochromatic light were able to recover normally. However, ciliary nerve section produced hyperopia, deepening of the anterior chamber and a tendency towards axial lens thinning, irrespective of the light conditions used. Hyperopic refractive errors peaked at 4 weeks (mean refractive errors: +5.7 D, +4.21 D for ML, WL groups respectively, 4 weeks), with the ML group still exhibiting significant hyperopia at 7 weeks. Ciliary nerve section did not prevent the myopic response to lid suture (mean refractive errors: -22.65 D; -25 D for ML, WL groups respectively, 1 week) nor the elimination of myopia when eyes were reopened. These data indicate that neither accommodation nor chromatic aberration are fundamental to the gross operation of the emmetropization process although they may be essential for the fine tuning of refraction. PMID- 8236849 TI - Light adaptation of human rod receptors: the leading edge of the human a-wave and models of rod receptor activity. AB - The human rod receptors can be studied by measuring the leading edge of the rod a wave of the ERG. Computational models, previously shown to fit the recordings from single rods, are fitted to dark-adapted a-wave responses. A model proposed by Lamb and Pugh [(1992) Journal of Physiology, 499, 719-758] fits slightly better than the traditional models based upon n-stage exponential filters. To test alternative models of rod light adaptation, a-waves were recorded to flashes presented upon steady adapting lights. Steady adapting lights decrease the rods' sensitivity. Human rods must adapt as response compression alone predicts far greater decreases in sensitivity. The evidence suggests that the mechanism(s) of adaptation include a change in the time-course of the rod's response. Human rods appear to adapt in much the same manner as do the rods of other vertebrates. PMID- 8236850 TI - Orientation, masking, and vernier acuity for line targets. AB - In an attempt to uncover the properties of the psychophysical spatial mechanisms which optimally respond to the vernier offset between two abutting lines, we investigated the effects of one-dimensional band-limited spatial noise masks superimposed with the target, on vernier thresholds. Unidirectional vernier thresholds were measured in the presence of masks varying in orientation, spatial frequency content and luminance modulation. Because of the dependence of vernier thresholds on target visibility, the effects of these masks on target detection thresholds were also measured. In accordance with the results of Findlay [(1973) Nature, 241, 135-137] but contrary to an hypothesis that the direction of the vernier offset is mediated by the differential output of spatial filters of a single orientation, our results reveal a bimodal orientation tuning function for vernier acuity. We propose that, for offset line targets, the differential responses of at least two filters with orientations which straddle the target lines are combined to extract relative position information. The spatial frequency tuning characteristics of the optimal mechanisms for mediating vernier information are similar to those optimal for detecting the target lines themselves, except that they are tuned to a slightly higher spatial frequency and have a slightly narrower bandwidth. The spatial mechanisms most sensitive to the vernier offset and to target detection exhibit similar responses to increases in mask modulation. This finding suggests that these tasks are limited by the same source of noise, and explains why under a variety of experimental manipulations, equally visible vernier targets result in similar vernier thresholds. PMID- 8236851 TI - Estimating the gradient direction of a luminance ramp. AB - We report on the extent to which human observers are able to indicate the gradient direction of a luminance ramp. In our experiment modulation depth ranged from 1 to 64% and field sizes subtended 0.5 to 47.5 degrees of visual angle. Observers are not able to indicate the gradient direction for modulation depths below 6%. For values above this threshold, the angular standard deviation in the responses decreases proportionally with the logarithm of the modulation depth and is about 11 degrees for a modulation depth of 64%. The angular standard deviations for supra-threshold contrast are slightly increased for the field sizes of 0.5 and 47.5 degrees but are constant over a range of field sizes of 1 to 25 degrees. Thus, size invariance holds well for this range of field sizes. PMID- 8236852 TI - Fusional suppression in normal and stereoanomalous observers. AB - Observers with normal stereopsis suppress some of the monocular information contained in each stereo half-image, a phenomenon we call fusional suppression. We measured vernier acuity for an ordinary vertical vernier test target, presented to one eye, that was paired stereoscopically with a vernier target with a large fixed offset, presented to the other eye. The size of the fixed offset, and hence, the disparity of the upper target line, was varied parametrically. Vernier thresholds for the test target rose as a function of disparity, reaching a maximum at 20 min of disparity and then decreasing gradually as the disparity exceeded the limits of foveal fusion (40-60 min arc). In the two normal observers, fusional suppression was symmetrical; neither eye had good access to monocular information. In the stereoanomalous observers, fusional suppression was not symmetrical. When they viewed the vernier test target with the stronger of their two eyes, their vernier thresholds were barely affected by the stereo half image in the other eye, and so were better than those of the normal observers measured in the same condition. When the stereoanomalous observers viewed the test target with their weaker eye, their fusional suppression was similar in range and magnitude to the suppression found in normal observers. Amblyopic suppression in mild amblyopes may be a residual effect of normal fusional suppression, operating to suppress monocular signals in the weaker eye, without conferring the benefits of normal stereopsis and fusion. PMID- 8236853 TI - The visual acuity for the lateral visual field of the pigeon (Columba livia). AB - Previous studies on pigeons indicated that the visual acuity for the frontal visual field was much higher than that for the lateral one. Suspecting that the poor values for the lateral field were due to suboptimal testing conditions, we determined the lateral field acuity in eight head-fixed pigeons with high contrast square-wave gratings. An instrumental conditioning task with water as reinforcer and mandibulation as an operant was used. Subjects achieved a mean acuity value of 12.6 c/deg. The results show that the acuity of the lateral visual field is only slightly lower than that of the frontal field. These data provide a psychophysical basis for ecological observations that pigeons and most other birds gaze laterally when scrutinizing small and distant objects. PMID- 8236854 TI - Development of color vision in goldfish: selective delayed maturation of blue vision. AB - Wavelength discriminations in juvenile and adult goldfish were measured among three sensitivity maxima (450, 525 and 625 nm) of goldfish cone photoreceptors using a "go/no-go" (electroshock avoidance) task. The ability of juveniles to discriminate was significantly poorer in 450/525 nm and 450/625 nm discriminations than that of adults. While only a few juveniles acquired clear discriminative responses, there was a greater proportion of adults. In contrast to those discriminations, the ability of juveniles to discriminate between 525 and 625 nm was similar to that of adults. These results suggest that juveniles, in contrast to adults, have selective delayed development of the blue-sensitive mechanism. Some possible explanations for the poor blue discrimination of juveniles are proposed. PMID- 8236855 TI - Neural recruitment explains "Weber's law" of spatial position. AB - We ask whether the well known Weber's law between spatial localization and element separation for high contrast, spectrally broad-band stimuli is a consequence of the organization of the early visual filters, or a fundamental constraint on the computation of spatial position by more central mechanisms. We address this question by identifying the individual contributions of mechanisms tuned to different ranges of spatial frequencies and contrast. We measure spatial alignment and bisection error as a function of element separation at each of a number of spatial scales, using spectrally narrow-band stimuli of fixed supra threshold contrast. We show that stimuli which minimize the extent of neural recruitment across different spatial channels before the site of extraction of the local contrast energy (and to a lesser extent across different contrast channels) do not exhibit Weber's law for either alignment or bisection. We present evidence that Weber's law for localization with increasing separation, found for stimuli of high contrast and broad-band spatial frequency content, is a consequence of the successive disengagement of unitary neural mechanisms, each of which has different spatial and contrast properties, and none of which individually exhibits Weber's law for spatial position. PMID- 8236856 TI - Spatial interactions in apparent contrast: individual differences in enhancement and suppression effects. AB - The apparent contrast of a small grating patch embedded in the center of an annular grating filled surround was studied as a function of the surround to central patch contrast ratio, the width of the annulus, and the contrast level of the central patch. Data from ten observers show large individual differences in apparent contrast, but these differences can be interpreted in terms of the relative strength of two interacting mechanisms. One mechanism mediates enhancement of the central patch. The other mediates suppression. PMID- 8236857 TI - The inverse intensity effect is not lost with stimuli in apparent motion. AB - The inverse relationship between the visible persistence of a briefly presented stimulus and its intensity is well established for static displays. However, with non-static displays, this relationship is only partially reported by previous studies. In order to clarify this topic, we investigated the effect of luminance on the visible persistence of a stimulus in apparent motion. Assuming that persistence duration is a normally distributed random variable, we studied whether the mean persistence of a stimulus could be systematically varied by varying its luminance. Our paradigm permits evaluation of this effect without changing the temporal interval between two successive presentations of the stimulus, thus avoiding the potential influence of this latter factor on persistence. Our results show that the inverse intensity effect still occurs at each of the successive locations of a stimulus in apparent motion. In addition, we provide evidence that increasing the spatial separation between the successive presentations, and decreasing the background luminance, result both in longer persistence duration. Altogether, these findings favour the hypothesis that persistence is actively suppressed by inhibitory interactions between adjacent neural zones. PMID- 8236858 TI - Simultaneous luminance contrast with chromatic colors. AB - The variations in the color of a test field of constant luminance during changes in the luminance of a contiguous inducing field was measured psychophysically. The fields had the same hue (red, green, or blue). The colors induced in the test field could be specified by the strength of a chromatic quality, and by the strength of the opponent qualities luminous/black. The psychophysical relationship between the two kinds of perceptive variables and test and inducing luminance followed distinctly different functions. The luminous/black variable varied linearly with contrast over a large range, as previously found for achromatic colors. The contrast gain of the luminous/black variable for the red and green colors was the same as for achromatic colors. The gain for the blue colors was twice the gain for the other colors. The chromatic variables were primarily related to the local luminance. For a given test luminance they were maximal near zero contrast. They followed the same function as the white component of achromatic colors. It is suggested that the luminous/black variable is related to spectrally broadband cells with a strong center/surround antagonism, while the chromatic variables and white are related to cells that lack a spectrally broadband surround in their receptive field. PMID- 8236859 TI - Shape from texture: ideal observers and human psychophysics. AB - We describe an ideal observer model for estimating "shape from texture" which is derived from the principles of statistical information. For a given family of surface shapes, measures of statistical information can be computed for two different texture cues--density and orientation of texels. These measures can be used to predict lower bounds on the variance of shape judgements of "ideal" and human observers. They can also predict optimal weights for cue integration for the inference of shape from texture. These weights are directly proportional to the information carried by each cue. The ideal observer model therefore predicts that the variance of subjects' responses in a psychophysical shape judgement task should reflect the statistical importance of individual texture cues. Our results show that human performance in shape judgements for a one-parameter family of parabolic cylinders is often better than what an ideal observer achieves using a density cue alone. Therefore other information, for example the compression cue, must be used by human observers. For the first time, such results have been obtained without recourse to the unnatural cue conflict paradigms used in previous experiments. The model makes further predictions for the perception of planar slanted surfaces in the case of wide field of view. PMID- 8236860 TI - Plan and operation: National Nursing Home Survey Followup, 1987, 1988, 1990. PMID- 8236861 TI - Questionnaires from the National Health Interview Survey, 1985-89. PMID- 8236862 TI - [Atrial natriuretic factor as a marker of clinical severity in cardiac failure]. AB - The authors examined plasma levels of the atrial natriuretic factor (ANF) in 26 patients with manifestations of cardiac failure. The ANF levels were high as compared with reference values (21 +/- 16.7 fmol/ml vs. 4.9 +/- 2.1 fmol/ml) and different in relation to individual functional NYHA classes, i.e. the lowest ones were in the second and the highest ones in the fourth class. Concurrently the authors revealed a statistically significant negative exponential correlation between plasma ANF levels and the left ventricular ejection fraction (r = 0.550, p < 0.01) and a bordeline correlation between ANF and the stroke volume (r = 0.45, p = 0.05). The results indicate that the plasma ANF level is a good indicator of severity of cardiac weakness and can be used in clinical work as a prognostic indicator in patients with heart failure. PMID- 8236863 TI - [New findings on the immunologic diagnosis of systemic lupus erythematosus]. AB - The authors review recent findings pertaining to the pathogenesis of systemic lupus erythematosus. It was revealed that knowledge of impaired humoral and cellular immunity is of great practical importance and substantially improves the prognosis of the disease. PMID- 8236864 TI - [Damage caused by monoclonal immunoglobulins and possibilities of therapy. 2]. AB - Monoclonal immunoglobulins (M-Ig) cause due to their chemical structure and physical properties some pathological manifestations. In the text all skin changes are analyzed which are supposed to be associated with M-Ig. Some light chains form amyloid masses which can damage almost all organs. M-Ig can perhaps enhance also the capillary permeability although their role in the capillary leak syndrome remains obscure. PMID- 8236865 TI - [The Saint Vincent Declaration--a call for increased care of diabetics.VI]. PMID- 8236866 TI - [The Saint Vincent Declaration and the Budapest invitation. VII]. PMID- 8236867 TI - [Linkage analysis in pedigrees with Alzheimer's dementia]. AB - Linkage analysis with three specific DNA markers was carried out on the Alzheimer's dementia pedigrees. The DNA markers used in our study, D21S1/D21S11, D21S13 and D21S16 in the 21q11.2-21q21 regions are localized. The results from different brain DNA samples and from peripheral blood samples of AD families were evaluated by two-point linkage analysis. The linkage between DNA markers and one AD pedigree was observed (D21S1/S11 and D21S13). PMID- 8236868 TI - [Polymorphisms in the pseudoautosomal regions of X and Y in DNA diagnosis]. AB - New findings on the pseudoautosomal area of X and Y chromosomes potentiate the importance of this part of the human genome for direct and indirect DNA diagnosis. The submitted paper is focused on the characteristic features of the given area with special emphasis on loci used in the Institute of Haematology and Blood Transfusion during DNA monitoring of patients after allogeneic transplantation of bone marrow and which thus provide important information with a major clinical impact. The authors indicate trends of future research focused on investigation of the polymorphism of the receptor of the haematopoietic factor GM CSF in relation to its function and the influence on the pathogenesis of AML. PMID- 8236869 TI - [BCR and ABL gene rearrangement in chronic myeloid leukemia]. AB - The fact of chromosome 9 and 22 translocation connected with the fusion of BCR and ABL genes occurring in 95% patients with chronic myeloid leukemia (CML) enables us to use molecular biology methods in CML diagnosis. By means of these methods we also can prove the rearrangement of BCR gene in some cytogenetically negative cases that are without so called Philadelphia (Ph) chromosome. 51 patients with myeloproliferative disease have been tested by Southern technique during the last year. The rearrangement of BCR gene was detected in 28 patients, in 13 of 14 patients where the Ph chromosome and also in 3 of 13 patients where the Ph chromosome was not detected. The detection of BCR gene rearrangement helped us to set the diagnosis of CML more precisely. PMID- 8236870 TI - [Lipoprotein (a): a genetic risk factor for early ischemic cerebrovascular stroke]. AB - In 45 subjects after a cerebrovascular attack (age less than 55 years) and their first degree relatives -45 siblings, 60 children, 9 parents and 31 partners- lipoprotein (a) levels by the ELISA method as well as some other lipids, lipoproteins and apoproteins were assessed and compared with two control groups (I and II) of healthy subjects with a negative family history of early atherosclerosis. The lipoprotein (a) levels were significantly higher not only in probands, as compared with the control group I (27.4 +/- 30.7 x 16.0 +/- 18.9; p < 0.05, Mann Whitney U-test) but also in their children when compared with control group II (28.1 +/- 31.9 x 12.5 +/- 16.3; p < 0.05). The lipoprotein (a) levels did not correlate with age, there was, however, a significant positive correlation with total cholesterol (r = 0.2244, p < 0.01), LDL-cholesterol (r = 0.2834, p < 0.0001) and apolipoprotein B (r = 0.1759, p < 0.05) when Spearman's correlation coefficient was used. Familial dyslipidaemia Lp (a) defined as lipoprotein (a) higher than 25 mg/dl (or 30 mg/dl resp.) at least in two close relatives in one family was found in 27% (and 21.6%) evaluated families (10/37 and 8/37 resp.). "Risk" levels of lipoprotein (a) > 25-30 mg/dl were found most frequently in combination with other types of hyperlipoproteinaemia--in subjects with types IIa and IIb, in families in combination with familial combined hyperlipidaemia. The results indicate that "risk" levels of lipoprotein (a) are a frequent genetically conditioned metabolic disorder in patients with premature ischaemic cerebrovascular attacks. PMID- 8236871 TI - [Immunogenetics and immunologic aspects of kidney and bone marrow transplantation]. AB - Progress in comprehension of the immunogenetics of the HLA-complex and the discovery of new very potent immunosuppressive agents have enabled organ and tissue transplantations to be performed as a relatively routine therapeutic method. Long-term outcome measured as a half-life in kidney transplantations is 25 years among HLA-identical siblings, 12 years in one haplotype-mismatched paternal donors, and 7 years in cadaver transplantation program. The long-term outcome in the latter group can be markedly improved--to as much as 19 years- when six-antigen program is observed (i.e. donor and recipient are identical in HLA-DR, -B, and -A antigens). The survival of patients after bone marrow transplantation (BMT) having as a donor a HLA-identical sibling has improved remarkably, too. However, only about 30 percent of patients who might benefit from a bone marrow transplant have a genotypically. HLA-identical sibling who could be a donor. Transplants from unrelated donors have become therefore an alternative method. To find HLA-matched donors the establishment of large registries are needed. One of them--Them Bone Marrow Donors Worldwide--in Leiden has over 1.2 million of potential donors at the time being (1992). PMID- 8236872 TI - [Chronic fatigue syndrome]. AB - The authors followed up for a period of 1-14 years 52 patients with CFS who met the criteria outlined by Holmes. The group comprised 10 men and 42 women. In 15% of these patients after a mean period of 5.5 years thyroiditis was diagnosed. Complete recovery was recorded in 20%, improvement in 32% of the patients, on average after 7 years. In the course of treatment mainly immunomodulating preparations were used. Indication of these drugs was individual based on immunological examinations. The success was only partial. The clinical condition of the patients did not correlate with serological findings of IgM, IgA and IgG antibodies against VCA nor with antibodies against EA of the EBV virus. PMID- 8236873 TI - [The means to improve the efficacy of treating light casualties in a troop unit]. PMID- 8236874 TI - [The organization of surgical care for light casualties at the medical evacuation stages]. PMID- 8236875 TI - [The organization of therapeutic care for those with mild illness and minor injuries]. PMID- 8236876 TI - [The prospects for improving specialized gynecological care for light casualties and those with mild illness among women service personnel]. AB - The article contains the analysis of work of gynecological units and female hospitals during the Great Patriotic war. The authors stresses the necessity to make an opportune scientific elaboration of the system of medico-evacuation procedures as far as the problem of medical care to service women is concerned in modern conditions. The paper makes a proposal concerning the creation of a gynecological unit inside the organic structure of a hospital for minimally wounded. PMID- 8236877 TI - [The delivery of medical care to burn patients who can return to duty, their treatment and rehabilitation at medical evacuation stages]. AB - On the basis of clinical and medico-statistic data the article describes the main medical and evacuation indices of slight or middle-severity burn injured. The paper contains characteristics of medical care, settles the patient holding period, discusses methods and indications for rehabilitation measures depending on residual pathological disorders. PMID- 8236878 TI - [The treatment of soft-tissue wounds by using sorbent bandages]. AB - On the basis of experimental and clinical researches the authors has found out that the sorption via wound using activated carbonic fibrous material if started at an early post shock period could be considered as an effective method of treatment of severe mechanic injuries with a vast wound surface. This method contributes to the favourable outcome of local wound healing and diminishes the number of complications mainly of purulent-septic nature. The positive influence of sorbent applied directly on wound after surgical treatment consists in promoting tissue metabolism, and reducing the risk of inflammatory processes. There are straight manifestations of general detoxicating effect: diminishing of catabolism processes, reducing of toxic metabolite concentration, stimulating of immune system, falling the rate of toxemia. As a result the number of complications has diminished 40%, and the number of lethal cases--15%. PMID- 8236879 TI - [The current approaches to the diagnosis and treatment of diabetes mellitus]. PMID- 8236880 TI - [The outlook for the use of radiodiagnosis for the status assessment and triage of light casualties at military medical installations]. AB - Roentgenological examination can considerably facilitate the selection of minimally wounded. It is possible to improve the efficiency of roentgenological diagnosis with the help of application of modern traditional roentgenological technique, as well as the usage of computer tomography and ultrasonic examination. There is a proposal to equip the hospital for minimally wounded with a AP 2Y1 mobile roentgenological complex, ultrasonic diagnosis cabinet, and X-ray unit for intraoperative examination. The posts of roentgenologist-specialist in ultrasonic diagnosis and roentgenological laboratory assistant if stated inside the organic structure of these hospitals, will make it possible to increase the hospital triage capacity up to 200 patient per day. PMID- 8236881 TI - [The evaluation of the economic efficiency of a system for rendering medical care to light casualties, their treatment and rehabilitation]. PMID- 8236882 TI - [The problems in the epidemic-control protection of light casualties]. PMID- 8236883 TI - [The problems of the field water supply for the troops and for military medical installations]. AB - The article discusses the questions of the improvement of field water supply of troops. There is a proposal to create the Service of field water supplies as an element of the Logistic Command subordination, increase the water ration, improve the system of quality water control, create a unique system of expert control, reinforce the units of water supply in field hospitals and hospital bases, make regulations for water regimen of personnel in hot climate regions. There is a conclusion that the problem of water supply needs to be considered by the Command and Research establishments of the Ministry of Defence of Russia. PMID- 8236884 TI - [The nature of the health-promoting measures during the rehabilitation of light casualties and those with mild illness]. PMID- 8236885 TI - [The delivery of medical care to therapy patients with mild illness in the Navy]. PMID- 8236886 TI - [The views of V. A. Oppel' on the organization of medical support for light casualties]. PMID- 8236887 TI - [The organization of the staged treatment of light casualties based on the experience of World War II and local prewar conflicts]. AB - On the basis of the experience of medical support of troops gained during pre-war local conflicts and the Great Patriotic war the authors make the analysis of the main stages in the development and formation of the system of treatment of minimally wounded in the tactical, Army and Front regions. The article describes a number of aspects which have scientific and practical significance for the improvement of the modern system of evacuation and treatment procedures. PMID- 8236888 TI - [The 25th anniversary of the Military Medical Department of the Central Institute for the Advanced Training of Physicians]. PMID- 8236889 TI - [The delivery of medical care to light casualties, those with mild illness and minor injuries among the missile forces]. AB - It is supposed that medical care to minimally injured and wounded will be rendered immediately at the zone of launching positions or command posts. Their evacuation to the medical establishments can be performed after combat launch of missiles or after adequate personnel replacement. Multipurpose hospitals of rocket troops will be able to render secondary medical care with the elements of tertiary medical care. Casualty staging must be performed by helicopters or ambulances to the military hospitals or to the rear hospitals of the Ministry of Health of Russia. The multipurpose hospital must have a gynaecological unit for minimally wounded women. PMID- 8236890 TI - [The primary statistical processing of the results of biomedical research]. PMID- 8236891 TI - [Experience with the use of personal computers in the military polyclinic]. PMID- 8236892 TI - [Methods for the preparation and performance of research in areas with emergency situations]. AB - On the basis of the experience of liquidation of medical consequences in the zones of emergency the authors came to a conclusion that organizational and clinical aspects of the Disaster Medicine had to be studied more profoundly, including the realization of scientific and practical researches immediately at the zones of crisis situations. The article contains methodics for scientific experiments, as well as various methods of work for scientists at the zones of emergency. This material can be used during organization and carrying out of tactico-special exercises with rescue teams and special medical units, and also for realization of scientific researches immediately at the zones of crisis, with further analysis of data in order to elaborate practical recommendations for the improvement of medical support of troops and civilian population during calamities and disasters. PMID- 8236893 TI - [A resuscitation aid in the first aid and medical care for the victims of severe trauma received in emergency situations]. AB - On the basis of their own experience in combat casualty staging, emergency situations and various publications the authors theoretically substantiate the necessity to reinforce the structures of pre-medical and medical care with reanimation teams which must be equipped with adequate assets capable to meet with emergency situations. The article contains information about training, equipment and organic structure of these teams. Taking into account the pathophysiological mechanism of severe injuries the authors give characteristics of the volume of medical care provided by these teams at various periods of casualty staging. The article stresses the necessity to combine the conduction anesthesia with central analgesia. The authors substantiate the necessity of the quest for a new analgetic as a drug of choice at the pre-hospital period, and describe its desired characteristic features: effective analgesia without breathing depression or circulatory iatrogenic disorders; long-term analgetic effect which gives the possibility to perform evacuation to the secondary care medical unit; possibilities for national fabrication of this drug in large quantities, impossibility of addiction to this drug. PMID- 8236894 TI - [The use of external fixation rod devices with a wooden outer framework in disaster medicine]. PMID- 8236895 TI - [The methodological principles of extracorporeal detoxication]. PMID- 8236896 TI - [The outlook for the development of drugs for preserving the combat readiness of those with minor injuries from ionizing radiation (a review of the literature)]. PMID- 8236897 TI - [The mechanisms of the injuries to the nerve trunks in gunshot wounds of the extremities (experimental research)]. AB - Two series of experimental researches were conducted on rabbits who had received gunshot wounds of the hind pads inflicted by middle and high velocity missiles. On the basis of the data obtained with the help of electroneuromyography, as well as light and electronic microscopy, it was found out the correlation of clinical, electrophysiological and pathomorphological parameters. Morphofunctional changes were disclosed not only in the proper zone of wound, but also at the upper and lower sides of sciatic nerve, as well as in central nerve system (SNS). The results obtained during these researches give the possibility to make a conclusion that coexistent lesions of nerves in gunshot injuries depend on the velocity characteristics of a missile and have compressive-tractive mechanism. "Central disorders" which appear in SNS exert a strong influence on the development of degenerative-regenerative changes. PMID- 8236898 TI - [The microsurgical procedure in the treatment of a solid intratruncal neuroma in the superior primary cord of the brachial plexus]. PMID- 8236899 TI - [Experience with the use of a radionuclide method in the diagnosis of diseases of the internal organs]. PMID- 8236900 TI - [The organizational and surgical aspects of treating light casualties at medical evacuation stages]. PMID- 8236901 TI - [The diagnosis and treatment of skin melanoma]. PMID- 8236902 TI - [The basic trends in the combined treatment of trophic ulcers of the lower extremities]. AB - The article summarized the experience of treatment of 253 cases of trophic ulcers of lower extremities (patients' age 18-82). Surgical method was the main one. Operative interventions were oriented at the main point of pathogenesis in order to close the ulcer defect. Considering the abnormalities of microcirculation as a common component of pathogenesis, the authors substantiate the prolonged arterial infusion via large arterial branches, which were catheterized in 76 patients. It is recommended to perform gnotobiological isolation, immobilized protease, pulsed flow, ultrasonic cavitation, laser irradiation. PMID- 8236903 TI - [The importance of epidemiological diagnosis in the prevention of natural-focus infections (based on work experience with troops under the conditions of a mountainous desert locale)]. PMID- 8236904 TI - [An analysis of the fatal outcomes from amebiasis in a mountainous desert locale]. PMID- 8236905 TI - [An evaluation of the effect of the factors from a radioactively contaminated locale on immunity]. PMID- 8236906 TI - [Nonspecific body resistance in servicemen with different nutritional statuses]. PMID- 8236907 TI - [The annual work capacity dynamics of flight personnel]. PMID- 8236908 TI - [The health status and morbidity in the crew members of submarines at different periods of combat training activity]. AB - It was found out in the result of complex examination of health status of 2020 enlisted men from 65 submarine crews of the North Fleet, that the inflammatory and infectious morbidity during the post-cruise period was considerably higher than during seagoing period. This morbidity was characterized by seasonal outbreaks. As for the types of the immune dependency, there were no seasonal changes. The peak of infectious morbidity coincides with the pre-cruise preparative period and post-cruise recreation due to "mixing" of the crews and diminishing of the non-specific resistance of an organism. PMID- 8236909 TI - [Ioakim Romanovich Petrov (on the centenary of his birth)]. PMID- 8236910 TI - [Automated information systems in the work of the army medical services of advanced foreign governments]. PMID- 8236911 TI - [The system of medical evacuation measures during the cleanup of the sequelae of natural disasters and industrial catastrophes]. PMID- 8236912 TI - [The system of medical evacuation measures during the cleanup of the sequelae of natural disasters and industrial catastrophes]. PMID- 8236913 TI - [The medical rehabilitation of injury and somatic patients]. PMID- 8236914 TI - [Changes in drug pharmacokinetics and pharmacodynamics under the influence of microwaves of different ranges]. AB - Human and animal experiments were conducted to compare the effect of microwaves different by their range on drug pharmacodynamics and pharmacokinetics. The response to the microwaves depended on the dose and site of the exposure, on the frequency of the electromagnetic modes. The results suggest that it is possible to use microwaves for potentiating and prolongation of some drugs action. PMID- 8236915 TI - [Speleotherapy in the combined health resort treatment of children with bronchial asthma]. PMID- 8236916 TI - [The differentiated treatment of obstructive bronchitis in children]. AB - Infants with bronchial obstruction (BO) received differentiated regimens of physiotherapy. Spastic BO was managed most successfully by a complex involving ultrasonic inhalation of broncholytics and sinusoidal modulated currents. In BO with a hypersecretory component the best results were achieved with combination of UHF electromagnetic field plus ultrasonic saline-alkaline inhalations given in 2 steps. PMID- 8236917 TI - [The effect of the siting of exposure to decimeter-range electromagnetic waves on the function of the hypophyseal-thyroid and adrenal systems in viral hepatitis]. AB - The author studied time course of changes in the thyroid and adrenal systems as shown by the levels of T3, T4, T3/T4, TTH, hydrocortisone, ACTH in response to exposure to decimetric microwaves (460 MHz, 30 mW/cm2). The microwaves were directed to the area of hepatic, thyroid, adrenal projections and combinations thereof. An optimal effect was achieved at the course exposure of the adrenal projection. PMID- 8236918 TI - [The effect of interference currents on the functional status and physical work capacity of athletes]. AB - The authors studied 90 patients going in for cyclic and acyclic sports. All of them were exposed to interference currents given according to the segmentary technique. It was established that such treatment promoted cardiovascular recovery as well as that of neuromuscular system, enhanced physical performance, corrected impaired levels of catecholamines. PMID- 8236919 TI - [The infrared radiation dynamics of the hand under the influence of physical loading in the rehabilitation of patients with tendon injuries to the flexor muscles of the fingers and the nerves of the hand]. AB - Infrared imaging of the hands has been conducted in kinesitherapy of patients with injured finger flexor tendons and hand nerves as compared to healthy subjects. A trend to normalization of the infrared images was recorded in the regimens without force loading, duration of the exercise schedule not less than 1 week, start of the treatment in the early postoperative period. PMID- 8236920 TI - [The fungicidal action of integral and selective ultraviolet radiation on yeastlike fungi of the genus Candida]. PMID- 8236921 TI - [The antibacterial properties of the mineral waters from the health resort of Anapa]. PMID- 8236922 TI - [The comparative evaluation of the efficacy of treating patients with chronic nonspecific lung diseases by the Respiron program and by traditional sanatorium health resort treatment at a central higlands health resort]. PMID- 8236923 TI - [The microcirculatory bed of the ischemic myocardium under the combined action of a low-intensity helium-neon laser and finoptin]. PMID- 8236924 TI - [The effect of a red laser and of an alternating magnetic field on the reparative processes after uranoplasty]. PMID- 8236925 TI - [The efficacy of the therapeutic-rehabilitative measures in humeroscapular periarthrosis in combined sanatorium-health resort treatment]. PMID- 8236926 TI - [The treatment of osteophytes of the calcaneus at a municipal balneological and pelotherapy clinic]. PMID- 8236927 TI - [Experience with the use of leech therapy at a health resort on patients with hypertension and ischemic heart disease]. PMID- 8236928 TI - [The structure and causes of nonpsychotic disorders in patients at a health resort]. PMID- 8236929 TI - [The assessment of functional changes in the cardiorespiratory system during the sanatorium-health resort treatment of patients with neurocirculatory dystonia]. PMID- 8236930 TI - [The choice of the treatment method for anterior rheumatoid uveitis in children at a health resort]. PMID- 8236931 TI - [The status of the selection and sanatorium-health resort treatment of children from areas of radionuclide pollution]. PMID- 8236932 TI - [The balneological classifications of radon therapeutic agents]. PMID- 8236933 TI - [The individual positions in the lecture by V. S. Ulashchik The general principles of the therapeutic-prophylactic use of physical factors]. PMID- 8236934 TI - [The role of the thyroid in regulating the functional activity of immunity and the possibilities for physiotherapeutic immunomodulation]. PMID- 8236935 TI - [The 2nd Bad Kreuznach Protocol (1992) on the status of research in the field of radon therapy]. PMID- 8236936 TI - [Elements of yoga therapy in the combined rehabilitation of myocardial infarct patients in the functional recovery period]. AB - Fifty-nine postmyocardial infarction patients received combined therapy involving chemotherapy, physiotherapy, therapeutic exercises and yoga therapy. Thirty-seven controls received the same treatment without yoga exercise. The yoga complex implied elementary simple positions, relaxation exercise and respiratory exercise. A clinical response evident in both the groups appeared more pronounced in the test group as shown by marked improvement in external respiration and blood counts, in exercise tolerance and psychosomatic condition of the patients. PMID- 8236937 TI - [The incidence of detecting influenza virus antigens in the peripheral blood lymphocytes of donors in relation to the epidemic activity of influenza viruses A and B]. AB - Antigens of influenza A and B viruses in the peripheral blood lymphocytes of normal human subjects are found regularly both in epidemic and interepidemic periods. The level of detection of viral proteins in lymphocytes varies widely and correlates with the epidemic activity of the viruses. Influenza virus antigens were found several months before a rise in the incidence of the disease, the per cent ratio of the identified antigens correlating with the pattern of antigen detection in nasopharyngeal washings during an epidemic outbreak. Most frequently, the antigen found in lymphocytes was that of the main etiological agent of a definite epidemic; less frequently the hemagglutinin of the virus accompanying the one dominant in a given epidemic was found. PMID- 8236938 TI - [Interference and its overcoming during the reproduction of 2 influenza A virus strains in an experiment]. AB - The interference between two influenza A virus strains was investigated in vivo. In mixed infection of chick embryos or mice two results were observed: maximal reproduction of both strains or interference which resulted in a reduction of reproduction activity, antibody production, or virulence of one of the viruses. No interference was observed upon inoculation of two strains apathogenic for the study host combined with an equal concentration of an infectious virus. PMID- 8236939 TI - [The isolation of the herpesvirus from yaks during the epizootic in Mongolia in 1990]. AB - Strains of virus belonging, according to the results of electron microscopic examinations, to the herpes virus family, were isolated in Vero cell culture from the organs and aphtha secretions in the buccal cavity of yaks developing the disease during an epizootic (more than 500 animals) in Khubsugul aimak (Mongolian village) in Mongolia, 1990. Virus-neutralizing antibodies were detected in the blood sera of the sick animals. The isolated strains differ from herpes viruses affecting man. No infection could be induced by materials of the sick animals in cows and oxen of different breeds in Mongolia. Seroepidemiological surveys are planned because of the mass character of the disease and its repeated occurrence. PMID- 8236940 TI - [The genetic characteristics of the Vergina serotype of the tick-borne encephalitis virus and its pathogenetic traits]. AB - The Greek Vergina strain of tick-borne encephalitis (TBE) virus was studied in comparison with 7 other strains by molecular hybridization of nucleic acids and by clinicomorphological markers of pathogenicity for monkeys and Syrian hamsters. By the genetical features the Vergina strain differed from the eastern and western TBE subtypes but was found to be similar to the strains of other subtypes of the Urals-Siberian, east-Siberian (Aina-1448) and Central Asian antigenic variant. This group of strains hybridized with cDNA at 65 degrees C only in the absence of 50% formamide, reacted with probe 1115 complementary to protein E gene, with 1-3 probes complementary to the conservative region of the genome but did not react with the probes corresponding to the variable regions of the genome. The Vergina strain is close to TBE genotype III. The Vergina strain was found to be virulent inducing subacute meningoencephalomyelitis which developed slowly and was accompanied by less marked morphological lesions in the cerebral cortex than those induced by the eastern subtype. The Vergina strain was demonstrated to persist in the brain, liver, spleen, and lymph node tissues. PMID- 8236941 TI - [The immunity indices in the infection of mice of different strains with the Machupo virus]. AB - The pathomorphological patterns and the activity of serum interferon, interleukin 1, tumor necrosis factor, natural killers, and proliferative activity of lymphocytes were studied in BALB/c and C57B1/6 mice intracerebrally infected with Machupo virus. The BALB/c mice showed 100% lethality at 8-9 days after inoculation while C57B1/6 mice were found nonsusceptible to Machupo virus inoculation by this route. The pathomorphological findings at the peak of clinical manifestations in BALB/c mice revealed no organ whose functional deficiency could lead to the death of the animals. Investigations of nonspecific immunity parameters revealed a direct dependence between their high activity and susceptibility of the animals to Machupo virus infection. It is assumed that the endogenous shock due to the high activity of immune response mediators is the cause of death in Machupo virus infection. PMID- 8236942 TI - [The antiviral action of medicinal plant extracts in experimental tick-borne encephalitis]. AB - Some mechanisms of inducing resistance to experimental infection with tick-borne encephalitis virus were studied in experimental mice treated with aqueous extracts of berries of Vaccinium vitis-idaea, black currant, Vaccinium myrtillus, and of greater celandine grass. The condition of the immune system organs (spleen and thymus) after treatment with the extracts under study was analysed. A correlation was found between the degree of developing resistance to infection, virus accumulation in the brain, blood, spleen and thymus and changes in some parameters (spleen and thymus indices) of these immunocompetent organs. Possible mechanisms of induction of resistance to virus by herb extracts are discussed. PMID- 8236943 TI - [The intermolecular disulfide bonds of the surface polypeptides in the infectious laryngotracheitis virus of hens]. AB - The envelope proteins of chicken infectious laryngotracheitis virus were investigated using immunoblotting, radioimmuno-precipitation, and diagonal SDS PAGE. Polypeptides with molecular weights of 205, 155, 115, 90, and 60 kDa were detected by rabbit antiserum and those of 205, 115, 90, and 60 kDa by chicken antiserum. When chicken antiserum was used, the 60 kDa polypeptide appeared to be the major immunogen. The 60 kDa polypeptide was shown to be a disulfide linked dimer with a molecular weight of 112 kDa. It is suggested that 60 kDa polypeptide is present in virions both in dimer (112 kDa) and monomer (60 kDa) forms. PMID- 8236944 TI - [The morphological changes in Ebola infection in guinea pigs]. AB - Ebola virus reproduction and morphological lesions were investigated in infected guinea pigs by electron microscopy. The liver was found to be the main target organ, whereas in other internal organs the pathological changes were insignificant. Ebola virus reproduction was demonstrated only in cells of the mononuclear phagocyte system. PMID- 8236945 TI - [The ultrastructural changes in guinea pig organs during the serial passage of the Ebola virus]. AB - Morphological study of internal organs of guinea pigs inoculated with Ebola virus at 2-8 passages was carried out. In the course of these passages the number of infected cells and virus particles in the organs was shown to increase, and the destructive changes in organs became more pronounced. In the 1st-3rd passages Ebola virus replication was observed in macrophagal cells only but beginning from the 4th passage of virus reproduction was found also in hepatocytes, spongiocytes, and fibroblasts. PMID- 8236946 TI - [The immunological indices in the intranasal method of revaccination with a live measles vaccine]. AB - Immunological parameters of the blood were studied in intranasal revaccination with a live measles vaccine (LMV) from the strain L-16. Alongside with specific immune response, LMV was shown to produce nonspecific changes in the system of immunocompetent cells the amount and duration of which were vaccine dose dependent. The immunomodulating activity of LMV administered by different routes was different. Especially marked was the difference in the degree of side effect of the vaccine on T-lymphocytes. A dose of 10,000 TCD50 in 0.5 ml of LMV produced sufficient antigenic stimulation by the intranasal route of revaccination being quite satisfactory from the point of view of immunological safety of measles vaccine. PMID- 8236947 TI - [A nonradioactive method for the immunoprecipitation analysis of HIV-1 antibodies]. PMID- 8236948 TI - [The quality of the screening for antibodies to the human immunodeficiency virus]. PMID- 8236950 TI - [The human T-cell lymphotropic virus type 1 (HTLV-1)]. PMID- 8236949 TI - [The detection and sequence analysis of the simian T-lymphotrophic retrovirus (STLV-1 Papio) by using the polymerase chain reaction]. AB - The gene amplification technique was used for detection and sequence analysis of STLV-1 Papio proviral DNA. The polymerase chain reaction was performed with a primer pair at tax region of HTLV-1, 7336-7354, sense strand, and 7516-7494, antisense strand. One microgram of DNAs isolated from LUG-4 cells and autopsies was used in a reaction volume of 50 microliters involving 30 cycles of amplifications. The reaction product was blunt-end cloned into pUC19 cut with Smal. The sequence was done with T7-polymerase using 32P-dATR as a label. Our results indicate that STLV-1 Papio provirus is actually present in the cells of a lymphoid cell line and tumor cells of lymphomatous monkeys. There are some differences between STLV-1 Papio and reported sequences of HTLV-1 and STLV-1. PMID- 8236951 TI - [Fermentation of engineered strain producing cholera toxin B subunit]. AB - Studies indicate that the cholera B subunit could be synthesized efficiently in corn steep liquor by engineered E. coli strain MM2. We have also determined the optical density, pH and the yield of B subunit kinetically during fermentation. Corn steep liquor medium has advantages in low cost, simplicity in technological process and high yield of B subunit which can reaches 40 micrograms/ml in 50L fermentative tank. PMID- 8236952 TI - [Morphogenesis of Bacillus sphaericus Ts-1 and formation of toxic proteins to mosquito larva]. AB - The close relationships between morphogenesis of Bacillus sphaericus Ts-1 and formation of toxic proteins were studied systemically and comparatively by Enzyme Linked Immunosorbent Assay, FITC-IgG Location and SDS-PAGE etc. This study showed that the toxic proteins of Ts-1 were produced in 8h culture at 30 degrees C. The toxicity increased steadily along with the sporification and then declined as the free spores increased greatly without collection immediately. It is suggested that substances homologous with crystal toxic proteins were situated on the cell walls. PMID- 8236953 TI - [Ultrastructural characteristics of Lyme disease spirochetes (Borrelia burgdorferi) isolated from Ixodes persulcatus in China]. AB - This paper analyzed morphological characteristics of Lyme disease spirochetes (Borrelia burgdorferi), isolated from Ixodes persulcatus from different areas of China, with scanning and transmissive electron microscopies. Cells of 8 strains were 8.4-36.0 microns long and 0.12-0.35 micron wide. There were 1-9 left-handed spires with wavelength of 1.09-4.30 microns and amplitude of 0.38-2.10 microns. Ends of the cells were pointed or spindle. 7, 8 or 9 flagella were inserted subterminally at each end of the cell. Inner structures of cells were no tubles. A few cells were at stage of division. These results showed that Ixodes persulcatus spirochetes possessed morphological characteristics of Borrelia burgdorferi and there were many types of morphological characteristics. PMID- 8236954 TI - [Studies on the mechanisms of resistance to trimethoprim in Salmonella typhimurium]. AB - The mechanisms of resistance to trimethoprim (TMP) in the clinical isolates of Salmonella typhimurium were studied. The experimental results indicated that the frequency of TMP resistance of 50 strains of Salmonella typhimurium was 76%. Seven of the resistant strains were highly resistant to TMP and four of them contained different plasmids which could transfer in the same species and between different ones, and could be eliminated with 8% of SDS. The results of comparing the activities and characteristics of dihydrofolate reductase (DHFR) in crude extracts from seven resistant strains and those from controls suggested that overproduction of the chromosomal DHFR was the resistance mechanisms in three plasmid-free strains and the levels of DHFR activity of the strains was increased during prolonged exposure to TMP in vitro. However resistance to TMP of the plasmid-containing strains resulted from production of plasmid-mediated TMP resistant type Ia DHFR and a new type DHFR which has not been reported up to now. The present work provided the theoretical basis for clinical use of TMP and attempts to suppress development and spread of duge resistance of pathogenic bacteria. PMID- 8236955 TI - Are women human? PMID- 8236956 TI - Ethnic pain styles in acute myocardial infarction. PMID- 8236957 TI - Premature infant-nurse caregiver interaction. PMID- 8236958 TI - Animals, care, and nursing research. AB - These multiple measures of health were positive and encouraging. It was possible to integrate nursing actions into protocols for this long-term study. It is the author's belief that by strengthening the humane care and handling of animals the study results can more accurately reflect the discipline of nursing. Through such efforts, discipline-specific findings could be strengthened when an animal model is used, particularly in long-term or survival nursing research. PMID- 8236959 TI - Ways of managing the experience of caregiving to elderly relatives. PMID- 8236960 TI - Measurement of self-perception of intuitiveness. PMID- 8236961 TI - Client advocacy and the moral orientation of nurses. PMID- 8236962 TI - Buffering nursing stress through play at work. PMID- 8236963 TI - Predictors of success on the newest NCLEX-RN. National Council Licensure Examination for Registered Nurses. PMID- 8236964 TI - CARIN system--database for Bayes' theorem applications. Computer-Aided Research in Nursing. PMID- 8236965 TI - Family scores revisited--a comparison of three approaches to data aggregation. PMID- 8236966 TI - Special issue: Fetal medicine. PMID- 8236967 TI - Amniocentesis and chorionic villus sampling. AB - Amniocentesis and chorionic villus sampling have been shown through prospective, multicenter trials to be safe and effective methods of prenatal diagnosis; accordingly, a knowledge of these tests is important for those physicians who care for women during their childbearing years. We review the indications, techniques, safety, accuracy, and efficacy of amniocentesis and chorionic villus sampling and compare the advantages and disadvantages of each diagnostic test. This review should enable physicians to provide appropriate counseling and information to women at increased risk for fetal abnormalities detectable by either of these procedures. PMID- 8236968 TI - Fetal tissue sampling--indications, techniques, complications, and experience with sampling of fetal skin, liver, and muscle. AB - Invasive prenatal testing has become an important way to evaluate fetuses at increased risk for hereditary disorders. In utero sampling of fetal skin, liver, and muscle may be required to diagnose before-birth disorders that cannot be diagnosed by analysis using chorionic villi or amniotic fluid. In the next few years, many of these conditions will be detected by DNA analysis, and the need for these procedures may decrease dramatically. First performed by fetoscopy, fetal tissue sampling is now most frequently done by inserting a biopsy needle under continuous ultrasonographic guidance. We describe the indications, techniques, complications, and experience with obtaining fetal skin, liver, and muscle biopsy specimens. PMID- 8236969 TI - Fetal ultrasonography. AB - Since its introduction in the 1950s, ultrasonography in pregnancy has been helpful in determining gestational age, detecting multiple pregnancies, locating placentas, diagnosing fetal anomalies, evaluating fetal well-being, and guiding obstetricians with in utero treatment. We review current standards and controversies regarding the indications, safety, accuracy, and limitations of ultrasonography in pregnancy. PMID- 8236970 TI - Cardiac ultrasonography in structural abnormalities and arrhythmias. Recognition and treatment. AB - Fetal cardiac ultrasonography has become an important tool in the evaluation of fetuses at risk for cardiac anomalies. It can both guide prenatal treatment and assist the management and timing of delivery. We recommend that a fetal echocardiogram be done when there is a family history of congenital heart disease; maternal disease that may affect the fetus; a history of maternal drug use, either therapeutic or illegal; evidence of other fetal abnormalities; or evidence of fetal hydrops. The optimal timing of evaluation is 18 to 22 weeks' gestation. An entire range of structural cardiac defects can be visualized prenatally, including atrioventricular septal defect, ventricular septal defect, cardiomyopathy, ventricular outlet obstruction, and complex cardiac defects. The outcome for a fetus with a recognized abnormality is unfavourable, with less than 50% surviving the neonatal period. Fetal cardiac arrhythmias are also a common occurrence, 15% in the series described here. Premature atrial or ventricular contractions are most commonly seen and usually require no treatment. Supraventricular tachycardia can result in hydrops and require in utero treatment to prevent fetal demise. Complete heart block, particularly in association with structural heart disease, has a poor prognosis for fetal survival. PMID- 8236971 TI - Preimplantation diagnosis. AB - Preimplantation embryonic biopsy and analysis offer couples at increased risk of having offspring affected with a genetic disorder the possibility of an early prenatal diagnosis. For many couples, this approach would avoid the issue of the selective termination of affected fetuses. Substantial advances have been made in the area of preimplantation diagnosis, but the possible difficulties with this approach cannot be ignored. PMID- 8236972 TI - Prenatal diagnosis using fetal cells from the maternal circulation. AB - All current methods of fetal karyotyping are invasive and carry a definite, albeit small, procedure-related risk. Because of this and testing costs, only women older than 35 years who have a greater risk for fetal aneuploidy are currently offered prenatal testing. But this detects only 20% to 25% of fetuses with Down syndrome. It would be a tremendous advance to find a noninvasive technique for prenatal diagnosis that carries no procedure-related risk and could be offered to all pregnant women. We describe a possible technique for noninvasive prenatal diagnosis that aims to identify fetal cells in the peripheral maternal circulation and successfully garner them for prenatal testing. Early attempts at fetal karyotyping were hampered by inaccurate diagnostic methods and cumbersome cell-counting techniques. Today, improved capabilities of identifying and enriching for fetal cells, coupled with sensitive methods of analysis such as the polymerase chain reaction, bring renewed enthusiasm to this task. Many technical issues, as well as serious questions regarding the test's utility, still exist, however, and must be explored and answered before the capture of fetal cells in the maternal circulation translates into reality for noninvasive prenatal diagnosis. PMID- 8236973 TI - Intrauterine transfusion with red cells and platelets. AB - Having direct access to the fetoplacental circulation by ultrasound-directed needle puncture has led to therapeutic interventions for fetal anemia and thrombocytopenia. Most cases of red cell alloimmunization associated with fetal anemia are caused by the antibody to the D red cell antigen. The intravascular transfusion of red cells to a hydropic fetus in such cases has notably improved survival. Nonimmune hydrops fetalis due to maternal parvovirus infection has also been treated successfully with the intravascular transfusion of red cells, whereas fetomaternal hemorrhage has not proved amenable to such therapy. Sensitization to the PLA-1 platelet antigen is the most common cause of fetal thrombocytopenia in maternal platelet alloimmunization. Fetal platelet transfusions have not proved to be a practical therapeutic modality for this disorder owing to the short half-life of the platelets. Platelets transfusions to the fetus just before delivery may avert the need for cesarean section in cases of severe thrombocytopenia. PMID- 8236974 TI - Fetal drug therapy. AB - Fetal drug therapy encompasses several areas, including the prevention of external genital masculinization in 21-hydroxylase deficiency syndrome (congenital adrenal hyperplasia), biochemical amelioration of methylmalonic acidemia, and biotin-responsive multiple carboxylase deficiency. The correction of cardiac arrhythmias has become relatively commonplace, and a reduction in the risks of neural tube defects is now possible with the use of preconceptual and early conceptual folic acid. Similarly, fetal function can be altered by the induction of fetal lung maturity using a number of agents; corticosteroids are the most common fetal pharmaceutic agent, and a number of other agents have also been tried. The most common route of administering pharmaceutic agents is through the mother and the placenta, although the direct administration of certain agents is becoming more common. PMID- 8236975 TI - Nonvascular needle and shunt placements for fetal therapy. AB - The nonvascular placement of needles and shunts for the in utero treatment of fetuses with fluid-filled, space-occupying anomalies has been done for about 10 years. The rationale for this approach is to attempt to prevent progressive impairment of organ function or lethal damage by early decompression. Experience has taught us that the key to success in these cases is the exclusion of associated anomalies and the use of appropriate tests to assess the residual organ function at the time of first diagnosis. In fetuses with hydrothorax, shunts can prevent pulmonary hypoplasia, and in those with obstructive uropathy, they can prevent the development of progressive lung hypoplasia and renal damage before a fetus is fully viable. In fetuses with ovarian cysts, prenatal puncture is occasionally indicated, but in those with hydrocephalus, the beneficial effect of prenatal drainage is more controversial. The catheters used for in utero placement have been improved and carry a smaller risk than open fetal surgical procedures. PMID- 8236976 TI - Fetal surgery. AB - While most prenatally diagnosed malformations are best managed by appropriate medical and surgical therapy after birth, a few may require or benefit from correction before birth. Extensive experimental work in animals and innovative technologic developments have made fetal surgery a viable approach to perinatal care. Managing preterm labor continues to be a vexing problem. At present, only a few life-threatening malformations have been successfully corrected. Additional research into the abnormal function of some malformations, the development of less interventional techniques, and fetal stem cell transplantation should lead to successful treatment of more fetal disorders. PMID- 8236977 TI - Scarless skin wound repair in the fetus. AB - The ability of a fetus to heal without scar formation depends on its gestational age at the time of injury and the size of the wound defect. In general, linear incisions heal without scar until late in gestation whereas excisional wounds heal with scar at an earlier gestational age. The profiles of fetal proteoglycans, collagens, and growth factors are different from those in adult wounds. The less-differentiated state of fetal skin is probably an important characteristic responsible for scarless repair. There is minimal inflammation in fetal wounds. Fetal wounds are characterized by high levels of hyaluronic acid and its stimulator(s) with more rapid, highly organized collagen deposition. The roles of peptide growth factors such as transforming growth factor-beta and basic fibroblast growth factor are less prominent in fetal than in adult wound healing. Platelet-derived growth factor has been detected in scarless fetal skin wounds, but its role is unknown. An understanding of scarless tissue repair has possible clinical application in the modulation of adult fibrotic diseases and abnormal scar-forming conditions. PMID- 8236978 TI - Cytogenetic analysis in prenatal diagnosis. AB - Chromosome analysis is the single most frequent test used in laboratory prenatal diagnostic studies. I summarize the current status of the field, including diagnostic problems in the laboratory and the clinical problems associated with communicating unexpected laboratory findings. I explore the effect of molecular genetics on these issues and its possible future effects on the entire practice of prenatal diagnosis as it relates to the risk for chromosome nondisjunction (trisomy). I also discuss the use of cytogenetic analysis in the prenatal diagnosis of certain inherited genetic diseases. PMID- 8236979 TI - Prenatal diagnosis and significance of fetal infections. AB - Viruses like rubella, cytomegalovirus, varicella-zoster virus and parasites like Toxoplasma gondii can be transmitted from a pregnant woman to her fetus and can affect fetal development. Several factors determine the likelihood of fetal infection and the risk of consequences for the fetus, such as the timing of transmission during gestation or the immunologic status of the mother. No single diagnostic modality can be applied to all infections. Knowledge of the diagnostic methods available is essential for accurate counseling and treatment of affected pregnant women. PMID- 8236980 TI - Prenatal diagnosis of inherited metabolic diseases. AB - Advances in the prenatal diagnosis of inherited metabolic disease have provided new reproductive options to at-risk couples. These advances have occurred in both sampling techniques and methods of analysis. In this review we present an overview of the currently available prenatal diagnostic approaches for the diagnosis of metabolic disease in a fetus. PMID- 8236981 TI - Clinical teratology. AB - The field of teratology has become increasingly important in preventive medicine programs. By avoiding specific teratogenic agents, many birth defects can be prevented. In this review we will summarize the currently documented teratogenic agents in humans. PMID- 8236982 TI - Ethical considerations in prenatal diagnosis. AB - Prenatal diagnostic testing raises a number of important ethical issues, some related to diagnostic testing in general and others related to the special circumstances of pregnancy. These issues are most effectively addressed in the context of a broader understanding of the goals of prenatal diagnosis. Our dual obligations--to the pregnant woman and to the fetus--have an important influence on the goals of testing. Testing seldom leads to treatment beneficial to the fetus, but more often can be beneficial to the pregnant woman, particularly if the information provided enhances her ability to make sound decisions about reproductive matters. The process of prenatal diagnostic testing can, however, limit a woman's sense of control over the decisions made about her pregnancy. It can also provide an opportunity for third parties to become involved in what are usually considered private matters. It is therefore important that the process of testing include adequate counseling and follow-up and that the patient's confidence be respected. As prenatal diagnostic technology expands, both in terms of patients to be tested and diagnoses to be sought, society will face difficult questions concerning access to testing and the justification for its use. PMID- 8236983 TI - Ethical issues in recommending and offering fetal therapy. AB - Fetal therapy should be offered and recommended for a viable fetus when these criteria are met: invasive therapy is reliably judged to have a high probability of being life-saving or of preventing serious and irreversible disease, injury, or disability for the fetus and for the child it can become; such therapy is reliably judged to involve low mortality risk and low or manageable risk of serious disease, injury, or disability to the viable fetus and the child it can become; and the mortality risk and the risk of disease, injury, or disability to the pregnant women is reliably judged to be low or manageable. When one or more of these criteria are not satisfied, intervention is experimental and can only be offered, not recommended, on the basis of benefit to future patients and the autonomy of the pregnant woman. PMID- 8236984 TI - Ethics of fetal tissue transplantation. AB - Now that the Clinton Administration has overturned the ban on federal funding for fetal tissue transplantation, old ethical issues renew their relevance and new ethical issues arise. Is fetal tissue transplantation necessary and beneficial? Are fetal rights violated by the use of fetal tissue in research? Is there a moral danger that the potential of fetal tissue donation will encourage elective abortions? Should pregnant women be allowed to designate specific fetal transplant recipients? What criteria should be used to select fetal tissue transplants? Whose consent should be required for the use of fetal tissue for transplantation? We review the current state of clinical research with fetal tissue transplantation, the legal history of fetal tissue research, the major arguments against the use of fetal tissue for transplantation, and the new postmoratorium ethical dilemmas. We include recommendations for guidelines to govern the medical treatment of fetal tissue in transplantation. PMID- 8236985 TI - [Return of sinus rhythm in patients with paroxysmal heart rhythm disturbances]. AB - The purpose of the work was an assessment of the time and mechanism of the return of sinus rhythm in patients with paroxysmal supraventricular heart rhythm disturbances, treated pharmacologically. The Holter method enabled a precise determination of the mechanism of sinus rhythm return. A group of patients could be isolated with bradycardia-tachycardia syndrome, in whom attacks of MAS syndrome occurred only after regression of arrhythmia. At the time of the return of sinus rhythm and immediately after it transient conduction disturbances and hyperexcitability may occur. PMID- 8236986 TI - [Evaluation of microsomal enzyme function in the liver in patients with duodenal ulcer treated with famotidine]. AB - The studies of microsomal liver enzyme activity by means of a respiratory test with 14C-aminopyrine (ABT) were carried out in 20 patients with duodenal ulcer treated for eight weeks with famotidine. The respiratory test was performed before the introduction of treatment (group I), after four weeks of treatment (group II), and after eight weeks of treatment (group III). The mean value of the respiratory test before treatment was 5.32% of the dose/h. After four weeks of treatment with famotidine a significant decrease was found of the test value (x = 4.79% of the dose/h). An even stronger inhibitory action of famotidine on the microsomal system was shown after eight weeks of the treatment (x = 4.46% of the dose/h). These results evidence a depressive effect of famotidine on the activity of liver monooxygenases. In patients taking this drug, other drugs metabolised in the hepatocyte microsomal system should be used with high caution, as well as those which are known to be inhibitors of this system. PMID- 8236987 TI - [Clinical effectiveness of acipimox and fenofibrate in patients with hyperlipoproteinemia type II]. AB - Hypolipaemic effectiveness and unfavourable effects of acipimox and fenofibrate were evaluated in patients with essential hyperlipoproteinaemia type II. The studies were carried out in two groups of 30 patients each. The studied patients were treated for three months with diet, and for another three months one group received acipimox (750 mg/day), and the other group was given fenofibrate (300 mg/day). After three months of pharmacological treatment in both groups a statistically significant decrease was seen of the concentrations of total cholesterol (by 13.8% and 19.3% respectively), LDL cholesterol (by 16.3% and 22.1%). TG (by 22.7% and 28.9%). The increase of HDL cholesterol concentration (twofold increase after fenofibrate) was non-significant. Desired LDL cholesterol values were achieved in eight patients (26.7%) treated with fenofibrate and in two patients (6.6%) receiving acipimox. Both drugs were well tolerated. Unfavourable effects were slight and did not require drug withdrawal. PMID- 8236988 TI - [Bleeding time in patients with chronic liver diseases]. AB - In 105 patients with chronic liver diseases, bleeding time was studied by Ivy method. A statistically significant difference was found in the bleeding time between patients with chronic hepatitis and controlled cirrhosis and patients with uncontrolled cirrhosis. In 10 out of 26 patients with prolonged bleeding time with platelet count over 100,000/mcl and prothrombin index value over 60%, clinical signs of haemorrhagic diathesis were observed. We believe that the evaluation of platelet function by measuring the bleeding time by Ivy method should enter the set of routine laboratory tests in patients with chronic liver disease. PMID- 8236989 TI - [Laparoscopic cholecystectomy--treatment outcome of 500 patients]. AB - 500 patients with symptomatic biliary stones disease have been treated by laparoscopic cholecystectomy (LCh). Contraindications, such as: acute inflammation, earlier laparotomies, common duct stones or obesity were considered as relatives. In cases with duct stones, ERCP with sphincterotomy and evacuation of duct stones was performed before LCh. Small percentage of LCh failures (2.6%) and of postoperative morbidity (3.4%) by undoubted advantages as: lack of postoperative paresis of digestive tract, reduced inability time for professional activity and low risk of postoperative abdominal hernia make this procedure attractive for patients and surgeons. PMID- 8236990 TI - [Results of treating inflammatory aneurysms of the abdominal aorta]. AB - The authors present the results of treatment of six inflammatory aneurysms in the material of 53 aneurysms treated surgically in the years 1987-1992. Direct results of surgical treatment, hospitalization period, and peri-operational complications are the subject of analysis. Peri-operational mortality of the patients with inflammatory aneurysm was higher than that of patients with non inflammatory aneurysm, undergoing elective surgery. Higher incidence of peri operational complications indicates higher operational risk of inflammatory aneurysm. PMID- 8236991 TI - [Effect of CRP preparation on serum lipids in patients with hyperlipidemia]. AB - In a group of 18 patients with essential hyperlipidaemia the influence was studied of multivitamin CRP preparation given for 12 weeks, on lipid peroxides, cholesterol and triglycerides in the serum. The level of lipid peroxides was decreased significantly. This effect was associated with the increase of the serum level of vitamin E. A decrease was also found of the levels of total cholesterol and LDL-cholesterol. PMID- 8236992 TI - [Comparison of effectiveness of adriamycin and epirubicin administered in the CHOP protocol to patients with malignant non-hodgkin's lymphoma]. AB - In a group of 30 patients with the diagnosis of malignant non-Hodgkin lymphoma of high histological malignancy, a prospective randomized study was carried out concerning the effectiveness and toxicity of the CHOP chemotherapy using adriamycin vs epirubicin. The effectiveness and tolerance of both treatment programmes are comparable. Good tolerance of the used epirubicin dose encourages to its increase in combined chemotherapy programmes in order to achieve higher therapeutic effectiveness. PMID- 8236993 TI - [Study on the effect of cefradine (sefril) on results of determining concentration of uric acid in serum]. AB - The divergence of the obtained results of uric acid determination in patients with proliferative diseases of the haemopoietic system who were given cefradine concomitantly induced the authors to carry out a more profound analysis of that problem. The serum uric acid concentration was determined by two methods. The method with phosphorotungstenic acid showed an effect of cefradine present in samples on overestimation of uricemia, while no such effect was noted when the enzymatic method was used. PMID- 8236994 TI - [Value of mammography and sonomammography in the diagnosis of benign breast diseases coexisting with genital pathologic lesions]. AB - In the paper the relationship was analysed between certain gynaecological diseases and benign changes occurring within the breast. The study was carried out in 400 patients and 108 females were subjected to the analysis. In 42 patients in this group histopathological examination was also performed. On the basis of the studies carried out among women in the Bydgoszcz region, a relationship was found between benign changes in the breast and certain genital diseases. PMID- 8236995 TI - [Analysis of direct causes of death of patients treated for glomerulonephritis]. PMID- 8236996 TI - [Computer-assisted laboratory diagnosis with graphic demonstration of results (system "DIAGEMES")]. AB - A DIAGENES computer system was developed which enables an doctor without experience in informatics to organise a database, to fill it with own diagnostic data, to verify this data from various standpoints, to assess the discriminative value of each of the used studies and the whole database with its eventual modification, as well as to develop an own clinically useful discriminating system. The result of the discriminative analysis has the form of a likelihood list or a computer figure, easy to evaluate. PMID- 8236997 TI - [Transforming growth factors and their role in chronic liver diseases]. AB - The transforming growth factors are endogenous polypeptide substances having own cell receptors. Among them two main factors have been isolated--transforming growth factor alpha (TGF alpha) and beta (TGF beta). TGF alpha is secreted by activated proliferating hepatocytes and by certain neoplastic cells. It stimulates the synthesis of DNA and migration of hepatic epithelial cells. In chronic liver diseases it is produced in increased amounts and stimulates then proliferation and regeneration. TGF beta is released by non-parenchymal liver cells, thrombocytes, and neoplastic cells. It inhibits the synthesis of DNA in many cells, inhibits the synthesis of albumins and fibrinogen and regulates the production of acute phase proteins. In chronic liver diseases it increases the production of fibronectin and collagen, inhibiting at the same time their enzymatic breakdown. TGF beta participates in the development and progression of hepatic fibrosis. PMID- 8236998 TI - [Principles and technique of multiorgan procurement for transplantation]. AB - On the basis of own experience from many years and literature model principles are presented of multiorgan obtaining (MOO) for transplantation. The paper describes the general principles of MOO and the basic elements of the technique of obtaining the heart, liver, and kidneys. The main purpose of the publication is to present the problems of MOO to wider circles of doctors in the context of striving to a unification of obtaining methods on the basis of the widely accepted European model. PMID- 8236999 TI - [Stab wound of the heart and liver]. PMID- 8237000 TI - [Coexistence of acute intermittent porphyria and Crohn's disease]. AB - A very rare coexistence is described of acute intermittent porphyria and Crohn's disease in a 46-year-old woman. The diagnosis was made on the basis of the overall clinical picture confirmed by laboratory tests (porphyria) and histopathological examination (Crohn's disease). PMID- 8237001 TI - [Symptomatic aneurysm of the abdominal aorta with coexistent horseshoe kidney]. AB - A case of the 66 years old patient with an expanded, symptomatic abdominal aortic aneurysm has been reported. During an urgent operation a horseshoe kidney lying in the front of aneurysm was detected. The kidney was mobilised and good access to the aneurysm was achieved without division of the renal isthmus. PMID- 8237002 TI - [Primary bifocal large bowel neoplasms]. AB - Two cases are presented of bifocal, synchronous colonic cancers. In both patients resection of the large bowel was done achieving good final result. The necessity is stressed of careful diagnosis ruling out the presence of multifocal neoplastic lesions in patients with suspected large bowel cancer. PMID- 8237003 TI - [The cattle tick Boophilus microplus (Canestrini, 1888) (Acarina, Ixodidae)]. AB - The external morphology and life cycle of Boophilus microplus--a cattle tick- were given. The control of ticks as well as host resistance to ticks were also mentioned. PMID- 8237004 TI - Description of Neobrachiella sp. (Copepoda, Siphonostomatoida, Lernaeopodidae) parasitic in the buccal cavity of California halibut (Paralichthys californicus). PMID- 8237005 TI - Haematological and morphological changes in experimental anisakiasis in pigs. AB - The aim of this paper is to establish the haematological and organic changes in halothane-sensitive and halothane-resistant pigs in the course of experimental anisakiasis. Experiments were carried out on two groups of pigs (3 animals each). The pigs from the first group were given fifty A. simplex B larvae, the pigs from the second one received ten larvae and then again fifteen larvae each after the 5th and 6th days. The number of leucocytes, neutrophils, lymphocytes, monocytes and eosinophils was greater--different in both series, but similar in halothane (stress)-resistance and -sensitive pigs. In the case of sensitive pigs much greater reactive changes were found in the stomach submucosa than in that of resistant pigs. In this group of pigs nematode larvae have also been traced in the submucosa of the same organ. PMID- 8237006 TI - [Long term survival of first stage larvae of Elaphostrongylus cervi (Nematoda, Metastrongyloidea) in a frozen state]. AB - Faeces of red deer from Bialowieza Forest containing first stage larvae of Elaphostrongylus cervi have been stored during five years in -20 degrees C. After this time 52% of the larvae survived. These larvae were fully alive and in experimentally infected snails Helix pomatia developed to the infectious stage. PMID- 8237007 TI - Preparation of the oncospheres of the cestode, Hymenolepis diminuta for experimental studies. AB - The paper deals with methods facilitating the preparation of oncospheres of the cestode, Hymenolepis diminuta, for experimental studies. Described in detail are procedures for the infection of the definitive hosts with the oncospheres; collection and artificial hatching of oncospheres; purification of hexacanths; preparation of extracts from the hexacanths; and preparation of hexacanths for electronmicroscopic studies. PMID- 8237008 TI - [Level of total protein and activities of selected enzymes in hemolymph of Lymnaea stagnalis (L.) (Gastropoda: Pulmonata) naturally infected with digenetic fluke larvae]. AB - Total protein and activities of: alpha-amylase, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase in haemolymph of the freshwater snail Lymnaea stagnalis, naturally infected with digenetic larvae were investigated. There were no any changes in these parameters in the snails infected with Cercaria tenuispina that occupies mainly hematocele sinuses. Significant increase of activities of all examined enzymes in haemolymph of the snails infected with larvae belonging to the Furcocercariae group was ascertained. These changes were proportional to the degree of injury of the digestive gland, examined with the use of the thymol turbidity test. Total protein level was significantly higher only in haemolymph of snails with higher values of this test. PMID- 8237009 TI - [Localization and activity of several enzymes in tissues of Bothriocephalus acheilognathi and Khawia sinensis tapeworms parasitizing carp]. AB - In this study, an attempt has been made to compare the most common species of tapeworms in carp: non-segmentary Khawia sinensis and segmentary Bothriocephalus acheilognathi on the basis of histoenzymatic examination of 8 enzymes, i.e. alkaline phosphatase, acid phosphatase, adenosine triphosphatase, nonspecific esterase alfa, succinic dehydrogenase, lactate dehydrogenase, leucine aminopeptidase and gamma-glutamylo transferase. PMID- 8237010 TI - [Nosema disease in apiaries of the Pila and Poznan provinces in 1983-1988]. AB - Samples of bees from the Pila and Poznan voivodships were examined in 1983-1988. Nosema apis infection was diagnosed in 30.9-92.6% of apiaries of Pila voivodship and in 40.1-91.7% of apiaries of Poznan voivodship. PMID- 8237011 TI - [In Memoriam Prof. Dr. Gerhard Piekarski (1920-1992)]. PMID- 8237012 TI - [Artificial respiration of premature infants]. AB - Better understanding of respiratory physiology and progress in ventilator technology have contributed to improved mortality and morbidity of premature neonates. Yet, pulmonary complications remain high and there is no consensus about the optimal regimen of mechanical ventilation. We report our satisfactory 10-year experience with conventional mechanical ventilation based on a relatively low incidence of pulmonary barotrauma. The introduction of surfactant has facilitated the ventilatory management of premature neonates since the usually rapid improvement of the acute lung disease after surfactant administration allows for earlier weaning from mechanical ventilation. However, our own results and the results from most surfactant studies show no significant reduction in the incidence of intraventricular haemorrhage. Thus, though mechanical ventilation and surfactant administration are milestones in neonatal therapeutic management, the problems encountered in very low birth weight neonates both with respect to mortality and morbidity have not been generally solved and underline the role of optimal perinatal management. PMID- 8237013 TI - [The psychosocial status of HIV-positive mothers and their children]. AB - We studied the psycho-social circumstances of HIV-positive women and their children, who were born between January 1, 1984 and June 30, 1991 and are being treated at the Department of Pediatrics. We interviewed 19 women out of 21 families using the problem-centered interview technique according to Witzel. 29 out of 30 children were older than 18 months. Four children were HIV-positive and the serology of one patient aged 12 months who had been perinatally exposed was yet unclarified. The study showed massive problems within these families, such as considerable partner problems, drug abuse and social isolation. Furthermore, there was a lack of child care to relieve the mothers. The predominance of drug problems and insufficient experience of social institutions with the problematic nature of HIV-infection in mothers and children require more efficient preventive measures. PMID- 8237014 TI - [Hemorrhagic complications after tonsillectomy and adenoidectomy. Experiences with 7,743 operations in 14 years]. AB - During 14 years the department of otolaryngology at Leoben performed 7743 tonsillectomies and/or adenoidectomies in children in cooperation with the department of pediatrics. 97 patients were treated for postoperative bleeding = 1.25%; one child died as a consequence of severe bleeding. The analysis of age, timing and severity of the bleeding includes 15 additional children whose surgery had been performed elsewhere. Only 7 of the 112 bleeding episodes occurred during the first 24 hours after surgery. Most cases (18) occurred on the fifth postoperative day. The latest episode occurred on the 20th day after tonsillectomy. Because of severe blood loss, 18 patients received a blood transfusion. Despite pre- and postoperative coagulation tests no patient suffering from a coagulation disorder was identified. Neither the coagulation screening nor a 48 to 72 hour postoperative in hospital observation could prevent the risk of postoperative bleeding. PMID- 8237015 TI - [Haemophilus influenzae type B (Hib) meningitis in a 14-month-old child despite two vaccinations with Hib conjugate vaccine (PRP-D)]. PMID- 8237016 TI - [Course and management of tetanus]. AB - Within a span of 18 months 4 patients (2 men, 2 women) were admitted with generalized tetanus to our neurological intensive care unit. The "period of onset" ranged from 2 to 14 days. All patients needed artificial ventilation over 17 to 38 days. The clinical course was complicated by gram-negative pneumonia in 3 of the 4 patients. Time of hospitalization ranged between 35 and 60 days. One patient had never been immunized against tetanus. The other 3 had not received a booster dose for over 10 years. All 4 patients survived, 3 of them without sequelae. PMID- 8237017 TI - [Comment on the contribution by Ernst, E.: Is prevention really better than healing?]. PMID- 8237018 TI - [Sinus node syndrome after orthotopic heart transplantation]. AB - The sick sinus syndrome after cardiac transplantation is a new entity characterized by impaired automaticity (bradycardia, pathologic function tests, latent sinus node dysfunction) or entire lack of sinus activity (junctional escape mechanism), while the sinus node refractoriness and AV nodal function is usually unaffected. It may significantly affect quality of life after cardiac transplantation. Postoperative sinus node dysfunction is usually transient. However, patients in whom sinus rhythm has not been established by the fourth postoperative week should undergo pacemaker placement, given the unreliability of escape mechanisms in the denervated state. Patients with a latent type of sinus node dysfunction are likewise at risk of developing severe symptoms. These patients who usually are not prophylactically supplied with a pacemaker, may benefit from aminophylline therapy. PMID- 8237019 TI - [Laparoscopic colon surgery--from experiment to initial clinical experience]. AB - The technique of Laparoscopic resection of the colon sigmoideum was practised in an experimental model before starting a clinical program of laparoscopic colonic surgery. Until now, two resections of the colon sigmoideum, one ileocecal resection, one right colectomy, one Hartmann's procedure and one transversostomy have been performed laparoscopically. Three patients had colonic carcinoma. One sigmoid resection had to be converted to open surgery and this patient underwent relaparotomy because of postoperative ileus. The postoperative course of the other patients was uncomplicated. The promising initial experience demonstrates that laparoscopic colonic surgery is feasible, although further studies have to prove advantages and indications, especially with respect to oncology. PMID- 8237020 TI - [The value of cytogenetic, molecular genetic and flow cytometry methods in diagnosis and prognosis of lymphoproliferative diseases after lung transplantation]. PMID- 8237021 TI - [Intravesical electrostimulation in detrusor hypocontractility]. AB - Intravesical electrostimulation activates bladder mechanoreceptors and can induce detrusor contraction. The latency of afferent impulses indicates that myelinated A-delta fibres are used for impulse transmission. Twenty patients, of whom twelve showed acontractile, and eight hypocontractile detrusor muscles, were treated by means of intravesical electrostimulation. Prior to treatment, the average residual urine was 218 ml. Eleven out of the 20 patients on completion of the stimulation programme had a residual urine of less than 100 ml. Only four patients remain on intermittent clean catheterization, in comparison with nine patients before electrostimulation was started. Bladder sensitivity improved in 75% and detrusor contractility in 40% of the patients. No adverse effects were seen. PMID- 8237022 TI - [Comparison of parallel and divergent augmented reconstructions of the anterior cruciate ligament]. PMID- 8237023 TI - [Comparative study of the anatomy of the temporomandibular joint and its imaging in magnetic resonance tomography]. PMID- 8237024 TI - [Helicobacter pylori: clinical pictures of an infectious disease with pathogen persistence]. AB - It is the aim of this paper to provide survey of the very fast developing field of medical research on Helicobacter pylori which includes as different branches as gastroenterology, pharmacology or microbiology. After examination of the several diseases, pathological mechanisms, the diagnostical techniques and the therapeutical regimens, the authors of this study favorize an early indication for eradication of helicobater pylori. Since research on Helicobacter pylori is divided into many fields, there is the attempt to come to a clear and more rational point of view concerning therapeutical strategies and management of the infection. It is another focal point to emphasize the important role that chronical gastritis induced by Helicobacter pylori plays as an important risk factor for gastric carcinoma. PMID- 8237025 TI - [Chance occurrence in illness]. AB - The chance as an objective existing category is responsible for the fact that the processes in the nature and in the society seem to be chaotic although they are running of determinated. The chance is of special significance for the behaviour of complex organized living systems. It is an essential factor for the evolution and the development of the variety of the species. Disease represents a being of the life and in this context the chance is of great importance, too. This is concerning life and death, the process of the pathogenesis of the disease and its cause as well the individuality in relation to the group, the population and the species. In all these processes the chance is of different significance, so one can speak from the dialectic of the chance. The chance also determines the borderline of our scientific cognition. PMID- 8237026 TI - [Annual occult blood tests lower intestinal cancer mortality]. PMID- 8237027 TI - New therapies for multiple sclerosis--when? PMID- 8237028 TI - Managed care is at the core of socialized medicine. PMID- 8237029 TI - Tiny infant article raises questions. PMID- 8237030 TI - Crack and aortic dissection. AB - Aortic dissection represents a medical and, potentially, surgical emergency. Hypertension and cystic degeneration of the media are predisposing risk factors in the pathogenesis. Sporadic reports of aortic dissection in association with drug abuse especially crack cocaine are now appearing. We present such a patient whom we recently treated at University of Wisconsin Hospital. PMID- 8237031 TI - Endometrial ablation as an alternative to hysterectomy. AB - Each year in the United States, more than 700,000 women undergo hysterectomy. Nearly half of these procedures are done for abnormal menstrual bleeding. There have been few alternatives available. Since 1988, endometrial ablation has become an alternative to hysterectomy for women who: choose not to, or cannot have, an hysterectomy; cannot, or do not wish, to use long-term hormonal therapy; or those patients who are unable to tolerate more involved surgical procedures due to additional medical conditions. PMID- 8237032 TI - Physicians, hospitals, and managed competition. The beginning of a new era in medicine. PMID- 8237033 TI - New MSMS service will help "level the playing field" in contract negotiations. PMID- 8237034 TI - PHOs enhanced health care delivery through physician-hospital cooperation. PMID- 8237035 TI - Managed competition: some concerns for physicians. PMID- 8237037 TI - DOT changes response to "no driving" reports. PMID- 8237036 TI - Insurance chief speaks out on health system reform. PMID- 8237038 TI - New claim form rules from OCI. PMID- 8237039 TI - A study on financial viability of clinics and physician practices in under-served areas of Wisconsin: summary of results with recommendations. AB - A survey of 21 physicians and clinic managers located in Health Professional Shortage Areas (HPSAs) and other under-served areas in Wisconsin found that to combat increasing costs, low reimbursement rates, and increasing charity care demands, several practices rely on outside sources of funding to remain financially viable. Also, the financial viability of physician practices in under served areas of Wisconsin is threatened more by the low reimbursement rates of Medicaid and Medicare than by the provision of charity care. Though few are limiting the number of Medicare and uninsured patients they will treat, many have begun implementing cost containment measures, including more strict collection policies. There are also indications of restricted access for Medicaid patients. PMID- 8237040 TI - WIPRO clarifies changes to quality review process. PMID- 8237041 TI - Prevalence and characteristics of weight loss attempts in Wisconsin, 1989. PMID- 8237042 TI - EVP report: the view from here thus begins the American health care odyssey. PMID- 8237043 TI - We like it here, but we're moving. PMID- 8237044 TI - How to save money in the medical system. PMID- 8237045 TI - Alcohol use during pregnancy: implications for physicians. AB - There has been concern since antiquity that the ingestion of alcohol by the pregnant woman could damage the fetus, but it was not until 1973 that the fetal alcohol syndrome was first described. The broad effect of alcohol-related birth defects, including fetal alcohol effects (FAE), has become apparent even more recently. Physicians can play a central role in the prevention of alcohol-related birth defects through early detection, education of the mother, and appropriate referrals. This paper reviews the effects of maternal alcohol use on the fetus, discusses the importance of assessing alcohol use among all women of child bearing age, and examines intervention strategies that physicians can use to help pregnant women stop or reduce alcohol use. Funding for this paper was provided by the Maternal and Child Health Block Grant. PMID- 8237046 TI - Control of supraventricular tachycardia with transcatheter ablative technique using radio frequency as the energy source. AB - Supraventricular tachycardia constitutes a significant clinical problem, frequently requiring medical intervention. We analyzed data in 413 consecutive patients with supraventricular tachycardia treated at our center with the catheter ablative technique and found the following outcome. Among 272 patients who had regular narrow QRS tachycardia without ventricular pre-excitation during sinus rhythm (Group I), control was achieved in all but one case (success rate 99.6%). Patients in Group II had ventricular pre-excitation during sinus rhythm and narrow or wide QRS tachycardia. Elimination of ventricular pre-excitation and control of the tachycardia was achieved in 104 of the 106 cases in this group (a success rate of 98%). Group III patients had atrial flutter-fibrillation as the predominate arrhythmia and control of rapid ventricular response was successfully achieved with atrioventricular junctional ablation in all 35 cases. Serious complications were related to anticoagulation and were seen in less than 2% of the cases. There were no instances of thromboembolic phenomena or death related to these procedures. It is concluded that the catheter ablative technique with radio frequency as the energy source is a highly effective means for control of supraventricular tachycardia with minimal associated risk of serious complications. PMID- 8237047 TI - The physician's role in the Wisconsin Department of Natural Resources disability permits. PMID- 8237048 TI - Deaths from nine major chronic diseases, Wisconsin, 1979-1988. PMID- 8237049 TI - House calls in the good old days: the stories of elderly Wisconsin physicians. PMID- 8237050 TI - Statewide Physician Health Program procedures. PMID- 8237051 TI - [Violence and health]. PMID- 8237052 TI - Intentional violence among adolescents and young adults: an epidemiological perspective. AB - Intentional violence consists mainly of non-accidental interpersonal violence and suicidal behaviour; the remaining categories are other violence and/or violence with undeterminted intention [corrected]. In almost all the countries considered in this study, intentional violence is taking on worrying proportions in adolescence (10-19-year-olds) and is on the increase among young adults (15-24 year-olds). The scale of the problem is relatively well known thanks to national mortality rates. The relative proportions of murders and suicides vary considerably from one country to another, though they remain fairly constant over time, whether the diachronic progression of mortality due to intentional violence increases, remains stable or falls in the country concerned; this is true for both sexes and for both the age groups considered (15-19 and 20-24). Nevertheless, in view of the much higher incidence of intentionally violent behaviour that does not result in death, more resources should be allocated to epidemiological studies in that area, especially in terms of quantitative and qualitative methods, where possible in association with interdisciplinary projects. Only with a better knowledge of the risk factors and if possible identification of the predictive factors (in the probabilistic sense of the term) could we devise, conduct and evaluate preventive measures that are better targeted than those used so far, whether the factors are socio-cultural, socioeconomic or psychosocial. This is the justification, especially the ethical justification, of further epidemiological studies of an analytical or even interventional nature, going beyond the descriptive studies that have been made by most researchers to date. PMID- 8237053 TI - [Comparative analysis of mortality due to violence in developed countries and in a few developing countries during the 1985-1989 period]. AB - A comparative analysis of crude death rates and specifically of deaths from violence was conducted for a large number of developed and developing countries which provide data to the World Health Organization's data bank. For these countries, the analysis shows, first of all, that violent deaths rank third among the major causes of death, after diseases of the circulatory system and malignant tumours, in most developed countries and in some developing countries with reliable data. The comparative analysis also reveals substantial variations in the level and structure of death rates, both among the developed and the developing countries. Thus the crude death rate varies by a factor of 2 in both sexes, both in the developed and the developing countries. For violent deaths, the ranges are even wide: the highest death rate is 3 to 4 times greater than the lowest, except for women in developing countries where it is only twice as high. The method reveals in which countries the situation regarding violent deaths is relatively tolerable and in which countries the situation is decidedly bad. Although it is difficult to draw up a clear classification whereby countries can be grouped according to their profile of deaths by cause, it can be seen that a number of countries have a fairly characteristic profile of mortality by cause; these are mainly the developed countries (Western Europe and other regions) and some developing countries (Hong Kong and Israel) with low crude death rates. In these countries certain causes of violent death predominate as a result of the level of development achieved; motor vehicle accidents, falls and suicides. Moreover, in the other developed countries (in Eastern Europe) and in the developing countries (except Hong Kong and Israel) there is a more "traditional" profile of mortality by cause; this profile is characteristic of the less developed countries where there are generally fewer deaths from suicide and from motor-vehicle accidents and where unintentional factors predominate over intentional factors. This analysis also brings out the very distinctive situation of the countries of Latin America with regard to violent deaths, particularly the large number of deaths from homicide in a number of countries. This is unquestionably a high-risk region for which further studies need to be undertaken, and where preventive measures need to be applied in order to curb the rising tide of violence and its harmful consequences for these societies.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8237054 TI - Violence against women: the hidden health burden. AB - Violence against women is a major health problem around the world. It often goes unnoticed and undocumented partly due to its taboo nature. A number of recent studies have explored the extent and patterns as well as the health consequence of violence in different cultures. The studies cited indicate that violence against women is widespread and an important cause of morbidity and mortality among women. Injuries due to violence have only recently been recognized as an important public health problem. More research is needed to improve our understanding of gender violence, and to design better interventions. PMID- 8237055 TI - Metabolic fate and disposition of 1,1,1,2-tetrafluoroethane (HFC134a) in rat following a single exposure by inhalation. AB - 1. The metabolic fate and disposition of [U-14C]-1,1,1,2-tetrafluoroethane ([U 14C]-HFC134a) has been determined in the male and female rat following a 1 h single exposure by inhalation to atmospheres of 10,000 ppm. 2. Of the inhaled dose, approx. 1% was recovered in urine, faeces and expired air postexposure indicating that absorption of this fluorocarbon across the lung is poor. Of this 1%, approx, two-thirds were exhaled within 1 h of the cessation of exposure as unchanged HFC134a. The remaining radioactivity was exhaled as [14C]-carbon dioxide or excreted in urine and faeces as trifluoroacetic acid. 3. Carbon dioxide was the major metabolite of HFC134a accounting for 0.22 and 0.27% of the inhaled dose in the male and female rat, respectively. Urinary excretion accounted for 0.09% of the dose and faecal excretion 0.04% of the dose by both sexes. 4. Total metabolism measured as the sum of the radioactivities in urine, faeces and as carbon dioxide amounted to 0.34 and 0.40% of the inhaled dose in male and female, respectively. 5. There were no major sex differences in the rates, routes or amounts of radiolabel excreted. Analysis of a range of tissues at 5 days postexposure showed a relatively uniform distribution of radioactivity. There was no evidence for a specific uptake of HFC134a or a metabolite into any organ or tissue analysed, including fat. PMID- 8237056 TI - Metabolites of sodium selenite and methylated selenium compounds administered at cancer chemoprevention levels in the rat. AB - 1. The metabolism of orally-administered sodium selenite and five methylated selenium compounds was investigated in the female rat at dosages equivalent to those used in other studies for prevention of mammary cancer. Dimethyl selenide (DMSe) exhaled within 24 h following dosing was measured, along with inorganic and monomethylated (MMSe) forms of selenium plus trimethylselenonium ion (TMSe+) in urine. 2. MMSe was the dominant metabolite of selenite given at low levels (0.1 ppm in the diet), but excretion of DMSe and TMSe+ increased sharply when selenite dosage was increased to the chemopreventive range of 3 ppm dietary Se. When similar chemopreventive levels of mono-, di-, or trimethylated compounds were administered, the total quantity of methylated metabolites was greater than for selenite and the metabolite profile reflected the expected point of entry into the intermediary metabolism pathway; the major metabolites were MMSe from Se methylselenocysteine, DMSe from selenobetaine methyl ester, and TMSe+ from selenobetaine. However, the profile of metabolites provided clear evidence that the methylated selenium compounds underwent demethylation, as shown by the excretion of inorganic and MMSe. Selenium administered as dimethyl selenoxide was almost completely excreted and about 90% of the dose was recovered as DMSe, indicating that reduction was the major pathway. For TMSe+, about 10% of the dose was excreted as DMSe and 84% as TMSe+. 3. A low, non-toxic level of sodium arsenite (5 ppm As in the diet) that is known to modify differentially the anticarcinogenic activity of selenite and methylated selenium compounds did not modify the excretion of the methylated selenium metabolites. 4. It is concluded that high anticarcinogenic activity is associated with extensive excretion of methylated Se excretory metabolites, but high output of such metabolites per se does not necessarily lead to anticarcinogenic activity. The whole animal has extensive capabilities for interconverting forms of selenium, and retains significant amounts in tissues, complicating the interpretation of Se metabolism and anticarcinogenic action. Further research is needed on the forms of selenium present in tissues. PMID- 8237057 TI - Metabolic fate of a new dihydropyridine calcium antagonist, CD-349, in rat and dog. AB - 1. The metabolic fate of 14C-CD-349, a new calcium antagonist, was studied in rat and dog. 2. After oral administration of 14C-labelled drug in both species, the plasma levels of radioactivity reached maxima at 1-2 h and declined with elimination half-lives of 6-7 h. In both species, 71-85% of radioactivity was excreted in faeces and 17-27% in urine in 120 h. Biliary excretion in rat after oral doses amounted to 33%. 3. The low ratio of unchanged drug to total radioactivity in plasma suggested that CD-349 underwent rapid metabolism in both species. 4. Twenty-two metabolites were isolated and identified from dog urine and an incubation mixture with 9000 g rat liver supernatant. Principal routes of biotransformation of CD-349 were similar in both species, and involved: (1) oxidation of the dihydropyridine ring to the corresponding pyridine ring; (2) denitration of the nitrate ester; (3) hydrolysis of the carboxy ester to the carboxylic acid; and/or (4) oxidation of the side chain, although quantitative interspecies differences were observed. PMID- 8237058 TI - Effects of perfluoro fatty acids on peroxisome proliferation and mitochondrial size in mouse liver: dose and time factors and effect of chain length. AB - 1. Male mice were fed a diet containing perfluoro fatty acids of varying chain length (i.e. perfluoroacetic, -butyric, -octanoic and -decanoic acids) at different doses (0.02 or 0.1% w/w of diet) for different periods of time (2-10 days), and effects on liver weight, hepatic mitochondrial protein and hepatic peroxisomal palmitoyl-CoA oxidation, lauroyl-CoA oxidase and catalase were monitored. 2. The greatest effects were obtained with perfluoro-octanoic and perfluoro decanoic acids, while perfluoro acetic acid was inactive. The effects with 0.02% w/w of diet perfluoro-octanoic acid were at least as great as those observed with 0.1%. A more detailed dose-response investigation focused on perfluoro-octanoic acid revealed that maximal effects with this substance could be obtained with a dietary dose of 0.01% for 10 days and that significant changes were also observed with 0.001%. 3. Maximal effects with 0.02% w/w of diet perfluoro-octanoic acid were attained after 6-10 days of feeding. 4. As with other peroxisome proliferators, perfluoro fatty acids increase mouse hepatic peroxisomal fatty acid beta-oxidation more extensively than they increase catalase, thus increasing hepatic oxidative stress. 5. As with other peroxisome proliferators, perfluoro fatty acids increase mouse liver mitochondrial protein. This effect is due primarily to a redistribution of mitochondria from the nuclear to the mitochondrial fraction, caused by an apparent decrease in the mean size of hepatic mitochondria after treatment. PMID- 8237059 TI - Biological fate of sulphur mustard in rat: toxicokinetics and disposition. AB - 1. The toxicokinetics of sulphur mustard were studied after i.v. administration (10 mg/kg) to rat. 2. After i.v. administration, blood concentrations of sulphur mustard were best described by a two-compartment model with distribution and elimination half-lives of 5.6 min and 3.59 h, respectively. The apparent volume of distribution at steady state (Vdss) was 74.4 l/kg and total body clearance (Cl) was 21 l/h kg-1. 3. Unchanged sulphur mustard was still detectable in the systemic circulation 8 h after administration. Appreciable and long (96 h) accumulation of 14C was found in the systemic circulation, and significant high affinity of 14C-sulphur mustard for red blood cells. 4. The disposition of 14C was also investigated after i.v. injection of 14C-sulphur mustard to rat. Urine was the major route of excretion of sulphur mustard and/or its metabolites. Of the administered radioactivity 80% was excreted in urine over 96 h, the greater part in the first 24 h after administration. Residual 14C continued to be excreted until 4 days later. In urine, no sulpho- or glucuronyl-conjugates were detected. Of dose < 3% was recovered in faeces. PMID- 8237060 TI - Metabolism of Casodex in laboratory animals. AB - 1. Casodex, a non-steroidal antiandrogen, was eliminated primarily in faeces by rat, mouse, rabbit and dog. Rat, mouse and rabbit eliminated 20-30% of a single oral dose (8-25 mg/kg) in urine; only 3-4% was excreted in urine by dog (2.5 mg/kg). Oral absorption was about 80% in rat, mouse, rabbit and dog. 2. Most of the dose was recovered in 48 h from rat, mouse and rabbit. In rat, < 1% of the dose was exhaled as 14CO2 and < 1% remained in the carcass after 7 days. Recovery from dog was incomplete in 4 days but consistent with the long plasma elimination half-life of 7-7.5 days. Casodex was eliminated from rat plasma with a half-life of 17-21 h. 3. Examination of urine indicated extensive metabolism of Casodex and showed a marked species difference. In rat, mouse and dog, Casodex was cleaved at the amide to yield a carboxylic acid and an aromatic amine which subsequently underwent ring hydroxylation with sulphate conjugation. In rabbit, the major urinary metabolite was Casodex glucuronide, conjugated on the tertiary hydroxyl. 4. The major component in faeces of all species was unchanged Casodex; some hydroxy-Casodex was also observed in rat faeces. Analysis of rat and dog bile indicated that Casodex and hydroxy-Casodex were eliminated in bile primarily as glucuronide conjugates. PMID- 8237061 TI - Changes in the amounts of cytochromes P450 in rat hepatic microsomes produced by cyclosporin A. AB - 1. The effect of cyclosporin A, an immunosuppressive drug, on the levels of individual cytochromes P450 of rat liver was studied by immunoblotting with specific antibodies. Changes in the metabolic activities of the microsomal enzymes were similarly studied. 2. Total P450 was decreased by treatment with cyclosporin A for 17 days. NADPH-P450 reductase activity was also decreased by treatment with cyclosporin A for 10 days and decreased further after 17 days. 3. Cytochromes 2B2 and 2C6 concentrations were decreased by treatment with cyclosporin A for 17 and 10 days, respectively. Administration of cyclosporin A for 17 days also decreased the levels of the male specific cytochromes 2C11 and 3A2, and consequently decreased testosterone 2 alpha-, 16 alpha-, 2 beta- and 6 beta-hydroxylation activities. Shorter periods (7 and 10 days) of cyclosporin A treatment decreased the level of cytochrome 3A2, which metabolizes cyclosporin A. In contrast, the level of cytochrome 2C13 was unchanged. PMID- 8237062 TI - Effects of D-limonene on hepatic microsomal monooxygenase activity and paracetamol-induced glutathione depletion in mouse. AB - 1. D-Limonene, a monoterpenoid constituent of citrus fruit oil, blocks tumour induction by chemical carcinogens in laboratory animals, apparently by preventing bioactivation of procarcinogens and by enhancing conjugation of proximal carcinogenic metabolites. 2. Inhibitory effects of D-limonene were measured in vitro using cytochrome P450 isoform-specific substrates. D-Limonene inhibited p nitrophenol hydroxylase (pNP) activity in vitro in liver microsomes from acetone , phenobarbital (PB)- and beta-naphthoflavone (BNF)-treated mouse, and 7 ethoxyresorufin O-deethylase (EROD) activity in microsomes from PB- and BNF treated mouse. p-Nitrophenol and ethoxyresorufin are substrates for cytochromes P2E1 and P1A1, respectively. No inhibition of benzphetamine (BNZP) or aminopyrine (AP) demethylases by D-limonene was observed. 3. EROD, BNZP and AP activities in liver microsomes were increased 18 h after i.p. administration of D-limonene to acetone-induced mouse, while pNP activity was unchanged. The immunodetectable protein level of cytochrome P2B1 in non-acetone treated mouse was increased 18 h after D-limonene, with no differences in P2E1 or P1A1. 4. Acute D-limonene did not protect against paracetamol (acetaminophen)-induced depletion of liver reduced glutathione (GSH). A prolonged paracetamol challenge (0.6% diet for 10 days) elevated liver cytosolic GSH-S-transferase activity (GST) two-fold and decreased liver GSH to 46% of control values. Dietary D-limonene (1.0% diet for 10 days) maintained liver GSH concentrations at 92% of control values in the paracetamol-challenged mouse without altering GST activity. D-Limonene also increased liver GSH concentration (23%) in mouse fed 1.0% D-limonene alone. PMID- 8237064 TI - [Screening in the physicians practice--introduction to the topic]. PMID- 8237063 TI - [The role of Helicobacter pylori in development of ulcers]. PMID- 8237065 TI - [Screening of colorectal cancer: indications and recommendations for a practical procedure]. PMID- 8237066 TI - [Rational diagnosis of prostate cancer]. PMID- 8237067 TI - [Screening for malignant gynecologic tumors]. PMID- 8237068 TI - [Screening in arterial hypertension, coronary heart disease and heart failure]. PMID- 8237069 TI - [Screening in the physicians practice for detection of diabetes mellitus and hyperlipoproteinemia]. PMID- 8237070 TI - [Osteoporosis--are screening examinations in the physicians practice useful?]. PMID- 8237071 TI - [Diagnostic errors in symptoms of oncologic diseases]. PMID- 8237072 TI - [From general practice in expert assessment: in meningitis in infancy typical symptoms are often missing]. PMID- 8237073 TI - [Case from general practice (13): nocturnal asthma attacks]. PMID- 8237074 TI - [Racial health concepts reflected by the Dresden International Health Fairs 1911 and 1930]. PMID- 8237075 TI - [Antihypertensive therapy and vascular wall protection with special reference to calcium antagonists]. PMID- 8237076 TI - Lipoprotein metabolism and coffee intake--who is at risk? AB - Data from a representative health and nutrition survey of German adults (sample of 1073 women and 806 men) were used to investigate the relationship between coffee consumption and the concentration of cholesterol in serum as well as other lipoprotein constituents. For these outcome variables multivariate analyses were conducted separately for men and women. Differences in age, body mass index, smoking habits, use of oral contraceptives, physical activity, alcohol, fish, fat, milk and tea consumption were controlled for in the models. Interactions between coffee drinking behavior and smoking habits as well as between coffee and the use of oral contraceptives in their relationship with serum cholesterol were of special interest in the analyses. Higher coffee intake (> 400 ml/d) showed higher total cholesterol, LDL cholesterol and lower triglyceride rich lipoprotein (TRL) and triglyceride concentrations in serum compared to lower intake (< 200 ml/d). Smoking appeared to be an aggravating factor in these relationships. Results of the linear regression analysis demonstrated an increase of 1.66 mg/dL LDL-C per cup of coffee daily consumed for men and of 1.58 mg/dL for women. The combination of high coffee intake, smoking and no oral contraceptive use ever was associated with the highest total and LDL-C and lowest TRL concentrations in this population. The observed differences may be explained by an increase of lipoprotein lipase activity due to coffee consumption. PMID- 8237077 TI - Studies on the metabolism of metallothionein and alkaline phosphatase of adult rat primary hepatocyte cultures: role of fetal calf serum and agonists of the phosphoinositide cascade. AB - Adult rat primary hepatocytes maintained in DMEM/F12 (Ham) media were used as a model system for studying the role of fetal calf serum (FCS) and agonists of the phosphoinositide cascade in the metabolism of metallothionein (MT) and alkaline phosphatase (ALP). Experiments were performed both after a 24 h preincubation with FCS and with bovine serum albumin (BSA). Hepatocytes were treated with dexamethasone (DEX), zinc (Zn) and with the agonists of the phosphoinositide cascade A23187, 1,2-dioctanoyl-sn-glycerol (DiC8), 12-O-tetradecanoylphorbol-13 acetate (TPA), angiotensin II (AT), platelet activating factor (PAF), Arg8 vasopressin (VP) and were analyzed for MT and ALP activity in cell homogenates. Cell viability was evaluated by lactate dehydrogenase (LDH) liberation into culture medium, induction of tyrosine aminotransferase (TAT) through DEX and by trypan blue exclusion. Overall, cell viability was improved by the FCS pretreatment and by DEX. Exposure of hepatocytes to the established direct inducers Zn and DEX of MT resulted in a manifold increase in MT, independent of whether the cultures were FCS pretreated or not. The FCS preincubation produced a moderate elevation of ALP activity by stimulating cell viability. However, ALP was unaltered in response to Zn and DEX. None of the experiments conducted with agonists of the phosphoinositide cascade led to an elevation of MT and ALP. Only the incubation of hepatocytes with A23187 resulted in a concentration dependent significant decrease of MT and ALP. This observation was due to a cytotoxic effect of A 23187, displayed by LDH leakage and an increase in the number of cells stained with trypan blue. In conclusion, in primary hepatocyte cultures agonists of the phosphoinositide did not have an effect on the metabolism of MT and ALP. Previous in vivo results indicating alterations of Zn metabolism in liver, therefore seem to be caused by indirect systemic responses. PMID- 8237078 TI - [The effect of zinc depletion on the fat content and fatty acid composition of the liver and brain in forcibly fed rats]. AB - In the present work the influence of zinc deficiency on fat content and fatty acid composition of liver and fatty acid composition of brain of rats with a high food intake was investigated. Using the force-feeding technique the rats were fed 14.5 g food daily at days 1 to 4, and then 11.6 g food for later days. After 7 days the zinc-deficient animals had a fatty liver which was characterized by an increase in fat content (68%) and dry matter (23%). The amounts of lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, and oleic acid were also increased by 100 to 200% in the liver of zinc-deficient animals, whereas the amount of arachidonic acid was decreased by 29%. The amounts of phosphatidylcholine and phosphatidylethanolamine in the liver were not changed by zinc deficiency, but the fatty acid composition of these phospholipids was changed. The liver phospholipids of zinc-deficient animals had a decreased proportion of arachidonic acid, but an increased proportion of docosahexaenoic acid. In the zinc-deficient animals there also existed a positive correlation between the fat content in the liver and the ratio between linoleic and arachidonic acid in the liver and a negative correlation between the fat content in the liver and the amount of arachidonic acid in the liver. These correlations as well as the changes in liver fatty acid composition of zinc-deficient animals suggest that the fatty liver might be the result of a disturbed metabolism of linoleic acid. In contrast, zinc deficiency did not influence the fatty acid composition of brain. This means that brain is protected against the effects of short-term zinc deficiency. PMID- 8237079 TI - [Characterization of guinea pigs after adaptation to different high vitamin C supplies. 3. Microsomal cytochromes in the liver and kidney]. AB - The specific amounts of the microsomal cytochromes P-450 (P-454) and b5 and the amounts of microsomal protein in liver and renal cortex of guinea pigs depend on the extent of the vitamin C supply to which the animals are adapted (at least 6 weeks). In the case of low supply--with 5 mg vitamin C in 100 g food--which still permits to survive, the amounts of the cytochromes are decreased in both organs (an accumulative supply--every 3.5 days via stomach tube--causes no evident decreases). The reduced amount of cytochrome P-450 gives rise to a corresponding elongation of the sleeping time after injection of evipane; this agrees with other reports upon restriction of metabolic activities by marginal supply. But the amount of the cytochrome is induced as well in these animals by several injections of evipane, to the same level as in animals supplied with more vitamin C. Our early report on a considerable decrease of the specific amount of cytochrome P-450 in the liver by omission of vitamin C for 14 days proves correct only in this organ and when the guinea pigs were previously abundantly supplied with vitamin C. There is no corresponding decrease after an adaptation to medium and lesser supplies of vitamin C. The decrease of the cytochrome after abundant supply seems to be due to a diminished stimulation of its synthesis in connection with modifications in the hormonal control of the metabolism and, thus, is only indirectly connected with the lack of vitamin C. The effects of omission of vitamin C supply probably depend generally on the previous adaptation, at least in the liver. The considerable decrease of the cytochrome P-450 and the simultaneous reduction of the mitochondrial compartment raise the question of if a maximum supply with vitamin C is favorable resp. the physiological optimum. PMID- 8237080 TI - [The effect of highly unsaturated fatty acids on the parameters of lipoprotein metabolism and rheology during their administration to patients undergoing chronic hemodialysis treatment]. AB - Fifteen hemodialysis patients were treated with capsules of highly unsaturated fatty acids given in addition to their normal diet over a period of 6 months. At study start they received 12 capsules daily, each containing 105 mg eicosapentaenoic acid and 75 mg docosahexaenoic acid. There was a significant fall of triglycerides from an initial 521 mg/dl on average to 334 mg/dl. Total cholesterol was reduced from 266 mg/dl to 233 mg/dl with a continuous fall throughout the study period, ldl cholesterol being the main part, falling from 227 mg/dl to 192 mg/dl. These changes were accompanied by a reduction of apolipoprotein A1 from 118 mg/dl to 109 mg/dl and of B from 212 mg/dl to 204 mg/dl. Parameters of whole blood viscosity decreased significantly (viscous component eta' from 6.81 to 6.44 mPas--elastic component eta" from 1.85 to 1.53 mPas), while the plasma viscosity remained unchanged with 1.44 mPas. The improvement of flow properties results from adherence of omega-3 fatty acids to the phospholipids of the red cell membrane. The conclusion is that supplementation with fish oil in addition to normal food intake of dialysis patients contributes to an improvement of lipoprotein metabolism and flow properties. Thus, two risk factors concerning thrombosis and cardiovascular diseases can be positively influenced. PMID- 8237081 TI - [Lipid adjustment of alpha-tocopherol concentrations in plasma]. AB - Measurement of alpha-tocopherol concentrations circulating either in plasma or serum is the most common method of measuring the vitamin E status in men. The analytical determination with a suitable HPLC method produces precise and reliable results. The interpretation of the measured alpha-tocopherol values is complicated by the strong positive correlation with different blood lipids. A considerable misinterpretation of the data will occur if these confounding factors are not taken into account by a suitable adjustment. Different methods have been described in the literature which tried to control the confounding effect of blood lipids. These methods were not completely satisfying. Therefore, a mathematical regression model, based on residuals, is introduced and applied to the data of the VERA-study (Verbundstudie Ernahrungserhebung und Risikofaktoren Analytik) and the consequences of the lipid-adjustment of the tocopherol values are studied in further statistical procedures. Thereafter, lipid-adjusted tocopherol concentrations can be subjected to more extensive statistical evaluations without the requirement of special stratification. PMID- 8237082 TI - Impact of some commonly used Egyptian diets on plasma lipids profiles of rats. AB - Seven types of diets commonly consumed in Egypt were studied in hyperlipemic rats. Each of the seven diets were fed to a group of hyperlipemic rats. The diets contain white beans and whole wheat bread (Diet 1); cabbage and peas (2); spinach and carrots (3); whole wheat bread and bean sprouts (4); whole wheat bread, white beans and peas (5); white beans and carrots (6) and the last diet contains peas and carrots (7). The experiment continued for 5 weeks, at the end of which different plasma lipids and biological parameters were compared to hyperlipemic rats fed the control diet for 5 weeks. Results showed that plasma total cholesterol of rats of all groups showed significant decrease, except in rats fed diet 6 in which no significant change was noticed. Plasma total lipids of rats fed diet 5 decreased significantly. Plasma phospholipids of rats fed diets 3 and 7 increased significantly. Rats fed diet 3 showed decreased levels of plasma high density lipoprotein-cholesterol. Body weight gain of rats fed diets 1, 2 and 3 decreased significantly. Total food intake was noticed to decrease on feeding diets 1 and 6. Significant decrease was produced in food efficiency ratio of rats fed diets 1, 5 and 6. Feeding any type of the previous seven diets produced significant increase of feces weight. PMID- 8237083 TI - Miso from peas (Pisum sativum) and beans (Phaseolus vulgaris) of domestic origin. Fermented foods from agricultural products in Europe. II. AB - Miso is a fermented soybean paste widely used in Japan as a soup base or as a seasoning agent. Koji (cereal grains with the mold Aspergillus oryzae) serves as enzyme source. Peas (Pisum sativum) and beans (Phaseolus vulgaris) of German origin can be used as substitutes for soybeans in the preparation of miso-like products. The legumes (peas, beans and soybeans for comparison) are washed, soaked in boiled water, dehulled and cooked for 35 min at reduced pressure. After grounding the seeds are mixed with salt, koji and mugi miso as starter and incubated at 45 degrees C for 14 days. During fermentation the glucose content increases up to 8-10 days and subsequently drops down. Crude protein decreases during incubation while dry matter increases. The pH value of all three miso types decreases during the fermentation period. Most of the 40 test persons characterize odor and flavor of the three misos as aromatic or sour; pea miso is often recorded to have a sweet-like odor and flavor. A legume-like taste of the final products has not been recorded. PMID- 8237084 TI - [An analysis of the correlation between trait related anger variables of the STAXI and situational anger reaction in an anger experiment]. AB - In our study we investigated the relationship between anger variables, which are supposed to be stable, and situational variables measured in an experimental setting (EISI-experiment: Experimentally induced stress in dyadic interactions). The correlations of questionnaire data (STAXI: State-trait-anger expression inventory) with self-reported and observed emotions in the anger experiment are presented. Furthermore, we discuss emotional reactions also under the perspective of buffering effects of attributions, that were stimulated during a failure situation in our experiment. Based on the theory of Weiner, we examined the influence of attribution processes on emotional behavior. The effects of this variation are especially interesting under the aspect of the anger process during the experiment. Persons with a low anger-trait are more stimulated by attribution processes than are subjects with high anger-trait scores. Apparently persons with low anger-trait scores get angrier when their cognitive interpretations of the negative outcome is stimulated. These findings are theoretically discussed. PMID- 8237085 TI - [Level specific attention and dominance of global markers]. AB - The global (i.e., highest) level of a hierarchically constructed stimulus pattern can often be identified faster than any subordinate level (Navon, 1977). This phenomenon was studied in two experiments aiming to confirm an attentional explanation of it. The hypothesis tested was that the reaction times for identifying each level of a three-level letter should be equal in a condition where valid level-specific cues are used. Experiment 1 provided results that show reduced differences between the reaction times for each level, but the differences were not reduced to near zero. The amount of interference in a condition with different letters on the level to be identified and the context levels was reduced substantially by using valid cues. One possible reason that the reaction time differences do not approach zero could be the nature of the stimulus onset produced by the imperative stimulus. This may cause attention to be automatically focused at the global level. By using a method that reduces this onset substantially, the efficiency of the level-specific cues was increased in Experiment 2. Attention was set at a certain level before stimulus presentation by identifying the first letter at a certain level in a double identification paradigm. This resulted in almost identical reaction times when attention was preset at the level to be identified. The strength of the level-specific attentional focusing depended strongly on the distinctness of the element's perceptual segregation at a certain level. This indicates that level-specific attentional selection, too, can be regarded as object-based. PMID- 8237086 TI - [Effect of body position and food composition on esophageal motility in healthy probands]. AB - In this study we investigated the influence of body position and bolus consistency on esophageal motility. Esophageal motility was investigated during randomized swallowing (10 women, 10 men) of liquid (5 ml water) and solid boluses (piece of apple). Esophageal motility was altered by body position and bolus consistency: Contraction amplitude, duration, percentage of multipeaked and repetitive contraction and the resting pressure of the lower sphincter were significantly reduced and the relaxation duration of the lower sphincter was significantly prolonged in sitting as compared to supine position. With a food bolus compared to swallows of water we found significantly higher values for the contraction amplitude and the percentage of repetitive contractions, for the duration and for the percentage of multipeaked contractions at some orifices and for the relaxation duration of the lower esophageal sphincter. Propagation velocity and the resting pressure of the lower esophageal sphincter were significantly reduced with the food as compared to the water bolus. The alterations of esophageal motility by body position and bolus consistency has to be taken into account in clinical manometry. PMID- 8237087 TI - [Familial juvenile polyposis with predominant stomach involvement]. AB - Juvenile Polyposis (JP) is a rare disease that may be found anywhere within the gastrointestinal tract, almost most cases so far reported have involved the colon. It is a precancerous condition, with the subsequently developing carcinomas so far also being found almost exclusively in the colon. A familial form is found in 20 to 50% of the cases. The present paper describes a family in whom three members of the second generation developed massive JP in the stomach requiring partial resection of the stomach or gastrectomy. Three members of the first generation died of carcinoma of the stomach and a forth of carcinoma of the colon. A male member of the second generation was treated at the age of 38 years for a carcinoma of the colon; 16 years later, he underwent resection of the stomach for juvenile polyposis and the histological work-up of the surgical specimen revealed in addition, areas of dysplasia and early carcinomas restricted to the mucosa. PMID- 8237088 TI - A new fine needle for easier, single handed, ultrasound-guided biopsies, requiring less advancing forces into solid organs. AB - A fine punch needle with a new bevelled geometry for easy, single-handed, ultrasound-guided biopsies is presented. The needle requires a significantly lower advancing force compared with the commonly used punch needles of the Menghini type. Since, therefore, our needle penetrates the organs to be investigated easier, it enables the puncture of easily movable (stomach, gut) and/or solid organs (kidney) with the yield of sufficiently long punch cylinders. The first 196 biopsies were well tolerated and histological diagnosis could be made in 94% of the cases. PMID- 8237089 TI - [Smoking and gastrointestinal function]. AB - Aspects of gastrointestinal function pathogenetically related to peptic ulcer have been studied extensively. Even as to secretion of gastric acid and pancreatic bicarbonate a definite statement concerning the impact of smoking is difficult. Most of the available studies found that smoking increases acid secretion and decreases pancreatic secretion. This may, at least in part, explain the untoward effect of smoking in ulcer disease. Gastric emptying is slightly decreased by smoking. Gastro-oesophageal and duodenogastric reflux are obviously not influenced to a clinically relevant extent. With regard to the colon, we did not retrieve any relevant publications. In summary, in patients with ulcer disease there is one good reason more to discourage from smoking. PMID- 8237090 TI - [Gluten hypersensitivity--are sprue/celiac disease only the tip of the iceberg?]. PMID- 8237091 TI - [p53 and hepatocellular carcinomas]. PMID- 8237092 TI - [Cholinergic receptor of the parietal cell--fact or fiction?]. PMID- 8237093 TI - [Improvement in myocardial function and perfusion after recanalizing a chronic coronary artery occlusion]. AB - The purpose of the study was to evaluate the improvement of the left ventricular ejection fraction and myocardial perfusion after recanalization of chronic coronary artery occlusions. The patients were investigated by rest and exercise radionuclide ventriculography (25/31) and rest and exercise myocardial scintigraphy (22/31). The examinations were performed 3 +/- 1 days before and within 7 days and 4 months after recanalization. Exercise-induced chest pain was present in 77% (24/31) of the patients before, in 10% (3/31) after recanalization and in 23% (7/31) during follow-up. Six of the 7 patients with exercise-induced chest pain after 4 months developed restenosis in the former reopened coronary artery. The results of the exercise-ECG present that 71% (22/31) of the patients had ST-segment-depression before, 19% (6/31) after catheter-intervention and 26% (8/31) during follow-up. Six of the 8 patients with exercise-induced ST depression after 4 months had a restenosis in the former reopened coronary artery. Reopening resulted in an increase of global rest ejection fraction (EF) from 51 +/- 11% to 54 +/- 13% (p < 0.05) and sectorial EF from 56 +/- 17% to 61 +/- 21% (p < 0.01) after recanalization. After 4 months patients with excellent angiographic results still had an increased global and sectorial EF at rest (global: 54 +/- 9%, sectorial: 59 +/- 17%; n.s.). Patients with restenosis (note: no reocclusion) developed a decrease of global and sectorial rest EF (global: 49 +/- 14%, sectorial: 57 +/- 19%; n.s.).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237094 TI - [Improved left ventricular ejection fraction directly after transluminal coronary angioplasty]. AB - The purpose of this study was to evaluate if left ventricular ejection fraction (EF) at rest can improve already during the first 4 h after successful transluminal coronary angioplasty (PTCA). Measurements were carried out with the CardioScint LV Function Monitor Version 1.3. Comparison of ejection fraction values evaluated by the CardioScint Monitor revealed a good correlation with usual equilibrium radionuclide ventriculography (n = 28; r = 0.89). Also, intra- and interobserver variability was low (n = 26; r = 0.94 and n = 21; r = 0.92 respectively). Two hundred follow-up-measurements in eight patients over a time period of 4 h revealed a significant decrease of EF from 45 +/- 12% to 39 +/- 18% (p = 0.037) (reference group). This decrease was due to a continuous increase of background activity. The study population consisted of 14 patients with successful PTCA (dilatation from 85 +/- 7 to 34 +/- 9% linear vessel diameter). To evaluate the hemodynamic significance of the coronary lesion, left ventricular ejection fraction was registered during the balloon occlusion of the vessel. After dilatation, 25 EF measurements were carried out in each patient over the following 4 h (350 values). During PTCA, EF decreased from 52 +/- 16 to 37 +/- 9% (p = 0.0027). During the 4-h follow-up EF slightly decreased from 50 +/- 8 to 46 +/- 9% (p = 0.07).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237095 TI - [Right ventricular function in patients with rheumatic mitral valve stenosis. Effect of balloon mitral valvuloplasty]. AB - Postrheumatic mitral stenosis might cause impairment of right ventricular (RV) function due to both an increase in RV afterload and rheumatic myocardial disease. Therefore, we investigated in 19 patients with postrheumatic mitral stenosis and sinus rhythm right ventricular volumes and hemodynamics by a computerized thermodilution catheter during rest and supine bicycle exercise. In 14 patients the investigation was repeated within 2 days after balloon mitral valvuloplasty. Resting RV ejection fraction was decreased (43 (15-53)%, median (range)) and correlated significantly with RV end-systolic volume index (r = 0.90), stroke volume index (r = 0.77), RV end-diastolic volume index (r = -0.76), heart rate (r = -0.69), pulmonary artery resistance (r = -0.69), and mean pulmonary artery pressure (r = -0.68). RV end-diastolic volume index was 107 (81 200) ml/m2. Exercise induced a decrease of RV ejection fraction to 36 (13-48)% at 50 Watt (p < 0.001), while it increased RV end-diastolic volume index to 131 (78 231) ml/m2 (p < 0.001). Balloon mitral valvuloplasty improved RV ejection fraction at rest from 41 (15-47)% to 48 (39-55)% (p < 0.005) and from 30 (13-46)% to 43 (27-56)% during exercise (p < 0.005). The increase of RV ejection fraction after valvuloplasty was caused by an increase in stroke volume, but not by a reduction in RV end-diastolic volume. CONCLUSION: Depending on the increased RV afterload, RV function is markedly depressed in mitral stenosis. An immediate and almost complete improvement of RV function occurs with the reduction of RV afterload after balloon mitral valvuloplasty.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237096 TI - [Clinical long-term results after corrective surgery of tetralogy of Fallot]. AB - We studied 246 consecutive patients, mean age 11.9 +/- 6.7 years, with primary (n = 155) or secondary (n = 91) complete repair of tetralogy of Fallot (TOF) between 1961 and 1972. Prospective follow-up was complete and ranged from 18.1 to 29.3 (mean: 20.3 +/- 4.2) years. There were 46 operative and 21 late deaths. Cumulative survival was 0.76 +/- 0.03 after 1 year, 0.72 +/- 0.03 (10 years), 0.68 +/- 0.04 (20 years) and 0.63 +/- 0.05 (25 years). After 20 years of follow up, which was a follow-up time available for all patients, cumulative complication rates were 0.17 +/- 0.03 for documented ventricular tachycardias/fibrillation, 0.16 +/- 0.03 for right-heart failure, 0.13 +/- 0.03 for left-heart failure and 0.11 +/- 0.03 for infective endocarditis. Eighteen of the 21 late deaths were from cardiac causes: sudden (n = 9), infective endocarditis (n = 4), left-heart failure (n = 3), and right-heart failure (n = 2). The hazard for ventricular arrhythmias was inconstant and increasing with time from the initial operation. After 20 years of follow-up, the cumulative incidence of sudden death, documented ventricular tachycardia/fibrillation was 0.81 +/- 0.07. Younger age at surgery resulted in a significantly better long term prognosis (p = 0.03) with cumulative survival rates after 20 years being 0.90 +/- 0.06 (ages 1-9 years), 0.92 +/- 0.04 (10 to 14 years), 0.83 +/- 0.09 (15 to 19 years) and 0.69 +/- 0.11 for patients being operated beyond age 20. Twenty years following TOF repair 59.2% of the late survivors were in NYHA functional class I and 36.2% in NYHA II. PMID- 8237097 TI - [Persistent ductus arteriosus--is closure indicated also in asymptomatic adults with small ductus and minor shunt?]. AB - We hypothetized that conservative treatment might be justified in asymptomatic adults with a small persistent ductus arteriosus. The data of 100 patients (age, 18-72 years) who later underwent non-operative closure were retrospectively analyzed. Left- and right-heart catheterization as well as angiographic measurement of PDA-size had been performed in all patients. Thirty-five patients were asymptomatic and 65 patients had cardiac symptoms. Six patients had a history of endocarditis. There were no significant differences with respect to Qp/Qs ratio (1.69 +/- 0.45 vs. 1.57 +/- 0.32) and ductus-diameter (4.7 +/- 1.4 mm vs. 4.8 +/- 1.3 mm) between symptomatic and asymptomatic patients. In the six patients with endocarditis the diameter was 4.5-7.0 mm. Symptomatic patients had higher mean pulmonary artery pressures (22.1 +/- 9.7 mm Hg vs. 18.5 +/- 5.1 mm Hg; p < 0.05) and were older (48.4 +/- 14.3 years vs. 30.9 +/- 11.2 years; p < 0.001) than asymptomatic patients. Neither from the size of the ductus nor from the Qp/Qs ratio could the development of cardiac symptoms or the risk of endocarditis be predicted individually. From these data it might be concluded that closure should be performed even in asymptomatic adults with small ductus and insignificant left-to-right shunt. PMID- 8237098 TI - [Dose-response relationship of nisoldipine in patients with stable angina pectoris and stress-induced myocardial ischemia]. AB - The dose-effect relation of nisoldipine in 3 single oral doses (5 mg, 10 mg, 20 mg) on exercise induced ST-segment depression, blood pressure and peripheral vascular response was evaluated in 16 patients with angiographically proven coronary artery disease and stable exertional myocardial in ischemia. The study was conducted according to a single-blind, randomized, placebo-controlled protocol with intraindividual cross-over. Maximal ST-segment depression and ischemia-score (ST-segment depression, workload time) under a constant submaximal workload exercise-protocol were analyzed with blood pressure, heart rate and the index b/a of digital pulse-plethysmography. 2 hours after medication nisoldipine led to a dose-dependent reduction of ST-segment depression by 12% (n.s.), 29% (p < 0.05) and 43% (p < 0.01), respectively. 8 hours after medication the ischemia score decreased from 2.98 to 2.34 with the 20 mg-dose (p < 0.05). Systolic and diastolic blood pressure were reduced dose-dependent by 7%, 9%, and 14% with no change of heart rate at 2 h after medication. Digital pulse-plethysmography showed an acute reduction of the index b/a from 0.72 (placebo) to 0.63 (-12.5%, p < 0.01) with 10 mg and to 0.56 (-22%, p < 0.01) with 20 mg. Thus nisoldipine showed a dose-related antiischemic efficacy lasting for at least 8 h with the 20 mg dose. A small but significant peripheral vasodilatation 2 h after medication was observed. PMID- 8237099 TI - [Heart volume determination as an indication of rejection after orthotopic heart transplantation]. AB - Endomyocardial biopsy in cardiac transplantation is still the most exact test (gold standard) to confirm or exclude a suspected rejection. Up to now, all available noninvasive tests suffer from a low sensitivity and specificity. For a further reduction of biopsies early available noninvasive methods are desirable. In the data presented a comparison between histological findings and heart volume measurements is shown in 40 patients (112 histological findings) after orthotopic cardiac transplantation. In 46 cases with no evidence of histological rejection the echocardiographically or radiologically determined heart volume was 869 +/- 142 ml/1.73 m2 body surface area, compared to 1026 +/- 178 ml/1.73 m2 in 29 cases with mild cardiac rejection. In 37 cases with a moderate or severe rejection heart volume as 1094 +/- 181 ml/1.73 m2. The diagnosis of a moderate or severe rejection by biopsy is combined with a heart volume above 950 ml/1.73 m2 with a sensitivity of 78% and a specificity of 73%. Intramyocardial edema as well as an increase in right and left ventricular volumes due to heart failure may be the causes for heart volume increase in rejection. Thus, heart volume measurement which can be performed easily during routine echocardiography gives additional information about whether a rejection has to be assumed or can be excluded. Today, sensitivity and specificity of the test does not allow a reduction of biopsy frequency. Further investigations are necessary to analyze the course of heart volume changes during an episode of rejection. PMID- 8237100 TI - [Dynamic magnetic resonance tomography in aortic valve insufficiency: methodologic comparison with aortography and color Doppler echocardiography]. AB - To evaluate the usefulness of cine-MRI for the quantitative assessment of aortic valve regurgitation, 31 patients (54 +/- 15 years of age; 22 M, 9 F) and 10 normal volunteers underwent a multi-modality imaging protocol comparing cine-MRI with color Doppler echocardiography and contrast aortography. Twenty-one patients had aortic regurgitation with an associated transvalvular gradient, 10 patients had pure valve incompetence. Aortic insufficiency as assessed by the signal void from regurgitant flow on cine-MRI was best analyzed on transversal tomograms. Both the volume of the signal void caused by turbulence above a threshold velocity, and the ratio of the diameters of the regurgitant jet and the outflow tract (dAL-Jet/DLVOT) were found to correlate closely with the (Seller's) angiographic regurgitant score by r = 0.86 each (p < 0.001), and the color Doppler echocardiographic index by r = 0.74 and 0.89, respectively (p < 0.001). Cine-MRI failed to differentiate the angiographic grades I and II, however, clearly separated grades II, III, and IV in contrast to other non-invasive imaging modalities. Moreover, a semiquantitative index derived from cine-MRI allowed a rapid assessment of the severity of regurgitation, similar to color Doppler echocardiography and the semiquantitative angiographic Seller's score. Thus, cine-MRI volumetric evaluation of transvalvular flow turbulences provides a useful and reproducible means to quantify aortic regurgitation. It also allows serial atraumatic investigations as a diagnostic alternative to color Doppler examination in patients less suitable for echocardiographic evaluation and may prove helpful in monitoring the natural course of aortic valve disease. PMID- 8237101 TI - [The Commission on the Ethics of the Treatment of Animals]. PMID- 8237102 TI - [The place of epiphyseal-adrenocortical relations in the corrective regulation of behavior]. AB - In the norm between epiphysis and adrenal cortex the unstable functional relations are established. During pathologic enhancement of adrenocortical activity, that causes the stress and psychic depression-related disorders in an emotional sphere, the epiphysial effects assume a stable restrained character. The major epiphysial hormone, melatonin, under pathology starts limiting the secretion of glucocorticoids eliminating in addition the behavioral disorders. PMID- 8237103 TI - [A neurophysiological analysis of the mechanisms of neuroendocrine regulation in stress and under the antistress action of the delta sleep-inducing peptide]. AB - In the article one can find analysis of modern theories about mechanisms of neuroendocrine regulation under stress, influence of steroid hormones and neuropeptides (delta-sleep-inducing peptide) on the functional state of hypothalamic reticular limbic structures of the brain. Special attention is paid to the analysis of mechanisms of creation of integrative processes under aggressive-defensive behaviour and roles of biologically active factors in the regulation of the processes. PMID- 8237104 TI - [Hemispheric interaction during the processing of spatial information]. AB - As a result studying the features of single neurons of prestrial cortex (field 21) of a cat the interhemispheric differences are revealed. It is found that the sizes of receptive fields (RF) of the neurons of this region in the left hemisphere is 1.5 larger as in the right one. The RFs of the left hemisphere consist of zones (mainly 5-9) than the RFs of the right hemisphere which in turn consist of 1-4 zones. About 70% of neurons of the left hemisphere are invariant to a direction of the movement of stimulus, the similar number of the right hemisphere neurons are selective with respect to orientation of a stimulus. The study of the spatial-frequency characteristics of RTs of this region indicated that the left hemisphere neurons are tuned to more lower frequencies (0.05-0.89 cycle/grade) than neurons of the right hemisphere (0.11-3.5 cycle/grade). The study of an activity of the neurons of field 21 after commissurotomy has demonstrated that in this case the hemispheres are fully lost the possibility to receive information from contralateral semifield of the vision. The sizes of RFs are significantly less and become equal in the both hemispheres. The neurons of this region become selective to an orientation of stimulus both in the left and right hemispheres. On the basis of the results of clinical and psychophysical studies it is customary to assume that the two sides of the visual perception- concrete and abstract--to a certain extent are disintegrated between each other and are supplied chiefly by different hemispheres.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237105 TI - [Myocardial heterogeneity and cardiac arrhythmias]. AB - Analysis of publications according to which myocardium consists of heterogeneous elements functioning as the whole. Evolution of ideas about structural-functional organization of heart tissue is shown. The conception and model of cluster structural-functional organization of myocardium are formulated. On this model processes of circulation of stimulation under fibrillation of heart ventricles are analysed. PMID- 8237107 TI - [The neuronal network of the hippocampus: a morphological analysis]. AB - The use of new methods of morphologic analysis made it possible to detail significantly already known insights and to formulate the new ones concerning the hippocampal construction. This review discusses in detail the principles of structural organization of all intrahippocampal interneuronal connections and projections to the hippocampus from entorhinal cortex. Also, other afferent and efferent pathways and connections of polymorphic neurons of hippocampus are studied. Revision of the lamellar principle of hippocampal organization and involvement of inhibitory system in restricting an excitation propagation along the hippocampus is discussed. PMID- 8237106 TI - [Milk hormones and their physiological importance]. AB - The paper deals with an analysis of the recently published literature on hormonal composition of milk. The origin of milk hormones, the mechanisms of their penetration into internal medium of a newborn and effect on formatting the neuroendocrinic functions and development of the immunity in a postnatal ontogenesis are being studied. The question relating to a need of isolating the functional system "mother--nursing baby" and to long-term negative effect of the use of artificial milk mixtures instead natural breast milk is posed. The dependence of the "mother--infant" system upon environments is shown. PMID- 8237108 TI - Sequence analysis of the Ebola virus genome: organization, genetic elements, and comparison with the genome of Marburg virus. AB - Sequence analysis of the second through the sixth genes of the Ebola virus (EBO) genome indicates that it is organized similarly to rhabdoviruses and paramyxoviruses and is virtually the same as Marburg virus (MBG). In vitro translation experiments and predicted amino acid sequence comparisons showed that the order of the EBO genes is: 3'-NP-VP35-VP40-GP-VP30-VP24-L. The transcriptional start and stop (polyadenylation) signals are conserved and all contain the sequence 3'-UAAUU. Three base intergenic sequences are present between the NP and VP35 genes (3'-GAU) and VP40 and GP genes (3'-AGC), and a large intergenic sequence of 142 bases separates the VP30 and VP24 genes. Novel gene overlaps were found between the VP35 and VP40, the GP and VP30, and the VP24 and L genes. Overlaps are 20 or 18 bases in length and are limited to the conserved sequences determined for the transcriptional signals. Stem-and-loop structures were identified in the putative (+) leader RNA and at the 5' end of each mRNA. Hybridization studies showed that a small second mRNA is transcribed from the glycoprotein gene, and is produced by termination of transcription at an atypical polyadenylation signal located in the middle of the coding region. The predicted amino acid sequence of the glycoprotein contains an N-terminal signal peptide sequence, a hydrophobic anchor sequence, and 17 potential N-linked glycosylation sites. Alignment of predicted amino acid sequences showed that the structural proteins of EBO and MBG contain large regions of homology despite the absence of serologic cross-reactivity. PMID- 8237109 TI - Expression of poliovirus P3 proteins using a recombinant vaccinia virus results in proteolytically active 3CD precursor protein without further processing to 3Cpro and 3Dpol. AB - The expression of the poliovirus genome occurs by the translation of a single open reading frame to generate a long polyprotein which is subsequently processed by viral encoded proteases. The initial proteolytic cleavages result in the production of a P1 polyprotein which contains the capsid proteins, and the P2 and P3 polyproteins which contain proteins required for replication. The P3 polyprotein consists of the 3AB protein (containing the viral genome-linked protein, VPg), the viral protease, 3Cpro, and RNA polymerase, 3Dpol. To further study the expression and proteolytic processing of poliovirus P3 proteins in vivo, we have utilized recombinant vaccinia virus vectors to express nucleotides 5240-7400 containing the P3 region proteins of poliovirus. The P3 protein expressed from the recombinant vaccinia virus VV-P3 exhibited in vivo proteolytic activity as evident by processing of the polyprotein to generate the 3CD protein, consisting of a fusion between the 3Cpro and 3Dpol proteins. Further processing of the 3CD protein to 3Cpro and 3Dpol, however, was not detected in cells infected with VV-P3. Subcellular fractionation of VV-P3-infected cells demonstrated that the 3CD protein was present in both the soluble and membrane fractions. Finally, the 3CD protein expressed from VV-P3 was stable in cells co infected with VV-P3 and poliovirus and no further processing to 3Dpol was detected. These results are discussed with regards to in vivo studies which suggest that the 3CD polyprotein is not a precursor to 3Dpol in poliovirus infected cells. PMID- 8237110 TI - Identification in the sera from patients with advanced cancer of a factor which stimulates gene expression from human immunodeficiency virus type 1. AB - From the sera of patients with advanced cancer, a novel factor called SDF (serum derived factor) was partially purified. SDF was shown to stimulate transcription from the long terminal repeat (LTR) of human immunodeficiency virus type 1 (HIV 1) by transient CAT assay. It did not stimulate gene expression of various control promoters including Rous sarcoma virus, human c-fos, c-myc, c-H-ras and chicken beta-actin genes. The SDF preparation did not contain any detectable TNF alpha or TNF-beta, and differed in its physicochemical properties from TNFs. We concluded that SDF might be a novel factor associated with the clinical features of advanced cancer. It is speculated that SDF might have some role in disease progression of AIDS as well as in the development of the cachectic conditions in AIDS associated with malignancies. PMID- 8237111 TI - Molecular and biological characteristics of a novel HIV-2 isolate, HIV-2HOM. AB - In 1991 a new HIV-2 isolate (HIV-2HOM) was isolated first from a German individual most likely infected in West Africa in the beginning of the 1970s. The virus was isolated from both, the plasma and the peripheral blood lymphocytes of the patient by using OKT-3-stimulated cord blood lymphocytes. The recovered viruses could be further propagated on Jurkat cells and exhibited a broad cell tropism. Biochemical and antigenic properties of HIV-2HOM were examined by radioimmunoprecipitations. For a more detailed molecular characterization, a 1520 bp DNA fragment from the env gene and a 722 bp DNA fragment from the pol gene were amplified by polymerase chain reactions, cloned and sequenced. A comparison of both sequences to prototypic HIV-2 and SIV isolates revealed a close relationship to HIV-2ST. This strain originated from an asymptomatic Senegalese individual and is supposed to be of reduced pathogenicity. Taking into account genetic data, it may be assumed that HIV-2HOM and HIV-2ST are closely related strains with different growth characteristics and pathogenic features. PMID- 8237112 TI - Comparative analysis of the conserved region of the orthopoxvirus genome encoding the 36K and 12K proteins. AB - Genes encoding virus-specific proteins with molecular masses of 36 kDa and 12 kDa were mapped in HindIII-P and HindIII-U DNA fragments of vaccinia strain LIVP and ectromelia strain K-1 viruses, respectively, by hybrid selection of RNA to cloned DNA fragments followed by in vitro translation. The 36K translation initiation codon was detected in the HindIII-J fragment. The nucleotide sequences of corresponding genes from vaccinia, ectromelia, cowpox and variola virus genomes were determined. The 12K protein has similarity to mammalian glutaredoxins. The derived amino acid sequence of the 36K polypeptide was compared with the protein bank PIR. No homology was found between the 36K protein and known structures of proteins. The 36K protein genes of vaccinia and ectromelia viruses were cloned in pUR290, which led to the production of E. coli chimeric proteins, consisting of the sequence of beta-galactosidase and the viral protein on their C-ends. The chimeric proteins were shown to possess viral antigenic specificity. To identify the protein product of the 36K gene monospecific antisera to chimeric proteins were obtained. The late 36K protein is associated with virosomes but is not incorporated into the virions of orthopoxviruses. PMID- 8237113 TI - Attempts to purify a second cellular receptor for a coxsackievirus B3 variant, CB3-RD from HeLa cells. AB - A coxsackievirus B3 variant, CB3-RD, isolated on rhabdomyosarcoma (RD) cells is known to bind HeLa cells at two different receptor protein sites, HR1 and HR2. Since HR2 occurs in almost 50 fold excess of HR1 in HeLa cells, purification of HR2 was attempted, to obtain its partial N-terminal amino acid sequence and its further characterization. This study describes the purification of HR2 from octylthioglucoside solubilized HeLa cell membranes (HeLa-OTG) by preparative isoelectric focusing (IEF) followed by either preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) or affinity chromatography on immobilized receptor monoclonal antibody, RmcA (RmcA-agarose). IEF of HeLA-OTG showed that both HR2 and HR1 could be well separated by this technique and focused with peak maxima around pH 3.7 and 6.7, respectively. Both RmcA and CB3 RD recognized HR2 as doublet bands (60 kD major polypeptide and a minor 55 kD polypeptide) on electroblots under non-reducing conditions. Preparative SDS-PAGE of the pool of IEF fractions containing HR2 (IEF pool) and simultaneous elution of polypeptides from the bottom of the gel during electrophoresis, is shown to be a useful technique in purifying HR2 with only one contaminating polypeptide (65 kD). However, affinity chromatography of the IEF pool on RmcA-agarose yielded HR2 without any detectable contaminating polypeptide. A quantitative chemiluminescence assay was developed to estimate the amount of HR2 on HeLa cells and in solution, when dot blotted on polyvinylidene difluoride (PVDF) membranes and probed with RmcA. Assays revealed that about 1.2% of the total HR2 present on HeLa cells could be obtained by IEF followed by affinity chromatography. Efforts are continuing to obtain sufficient quantities of purified HR2 for partial N terminal amino acid sequencing. PMID- 8237114 TI - Alterations of the stalk of the influenza virus neuraminidase: deletions and insertions. PMID- 8237115 TI - [International cooperation on the detection of irradiated food]. AB - A survey over recent international developments to detect the irradiation treatment of foods is given, in particular the programmes of "ADMIT" (FAO/IAEA) and of BCR (European Community). The need to detect radiation treatment by analysing the food itself is desirable to check compliance with existing regulations, such as the enforcement of labelling and control of prohibition, to enhance consumer confidence in the correct application of radiation processing, and to protect consumers' freedom of choice between irradiated or unirradiated food products. Some larger collaborative studies on an international scale have already taken place, e.g. ESR measurements of bones from chicken, pork, beef, frog legs and fish, thermoluminescence of insoluble minerals isolated from herbs and spices, gas chromatographic analysis of hydrocarbons and alkylcyclobutanones derived from the lipid fraction of chicken and the microbiological DEFT/APC procedure for spices. These methods could soon be implemented in international standard protocols. PMID- 8237116 TI - Extraction and determination of biogenic amines in fermented sausages and other meat products using reversed-phase-HPLC. AB - A convenient method is described for the analysis of biogenic amines (BA) by means of reversed-phase-HPLC. The method is characterized by multi-channel UV detection (diodearray), subsequent post-column derivatization with o phthaldialdehyde and 3-mercaptopropionic acid, and fluorescence detection. For the analysis of meat products and especially fermented sausages an optimized perchloric acid extraction process was introduced to determine putrescine, cadaverine, histamine, tyramine and 2-phenylethylamine. BA recoveries from meat ranged between 96 and 113% with a detection limit for amines of 0.5 mg/kg. PMID- 8237117 TI - [Is there a correlation of dietary fiber to the calculation of calorific value of food? Comparison of the experimental determination and the calculated metabolizable energy of 16 diets with different fiber content]. AB - The metabolizable energy (ME) of 16 diets containing various amounts and sources of dietary fibre was calculated by different energy assessment procedures and determined experimentally in balance studies in humans. ME was calculated by the German procedure (section 35 LMBG; 4/9/4) and by the British procedure (4/9/3.75). In addition, both calculation procedures were modified to take into account the energy value of dietary fibre. Dietary fibre was multiplied by an energy conversion factor of 1 or 2 kcal/g, respectively. The best agreement between measured and calculated ME was obtained when a conversion factor of 1 kcal/g fibre (German procedure) or of 2 kcal/g (British procedure) was used. PMID- 8237118 TI - Determination of flavone C-glycosides in tea. AB - An HPLC method for the determination of flavone C-glycosides (FCG) from black tea has been developed. Sample clean-up was accomplished by means of polyamide column chromatography, followed by enzyme hydrolysis of interfering compounds such was flavonol glycosides and a second polyamide column chromatographic step. Using HPLC with gradient elution and photodiode array detection eight FCG were separated. Seven FCG were isolated by means of preparative HPLC. Identification was carried out using co-chromatography, FAB(Fast Atom Bombardment)-mass spectrometry and various nuclear magnetic resonance techniques. Apigenin 6-C glucosyl-8-arabinoside (schaftoside) and apigenin 6-C-arabinosyl-8-C-glucoside (isoschaftoside) as well as luteolin 8-C-glucoside (orientin) and luteolin 6-C glucoside (isoorientin) have been detected in tea for the first time. Three of the other compounds have been identified as apigenin 8-C-glucoside (vitexin), apigenin 6-C-glucoside (isovitexin) and apigenin 6,8-di-C-glucoside (vicenin-2). Their occurrence in tea has been previously reported. From its UV spectrum another compound was concluded to be an apigenin glycoside. The FCG were quantified in a variety of teas of different origins (16 black, two green and one oolong). The total amounts of the FCG were 0.48-2.69 g/kg dry weight. The FCG pattern of teas of different origins were similar to each other and no origin dependent characteristics have yet been observed. Small amounts of FCG (1.2-2.2 mg/kg) were detected in hydrolysates of high relative molecular mass fractions (Mr > 5000) of a black tea liquor. PMID- 8237119 TI - Photodegradation of the carbamate insecticide pirimicarb. AB - In order to study the photoreactivity of pirimicarb (2-(dimethylamino)-5,6 dimethyl-pyrimidin-4-yl-N-dimethylcarbamate ) on plant surfaces, model experiments with organic solvents were performed. Pirimicarb decomposed readily when irradiated (lambda > 280 nm) in the presence of the selected model solvents to form a number of products. Half-lives were in the order isopropanol < cyclohexane < cyclohexene (up to a maximum of 140 min). In the presence of all three solvents photooxidation predominated leading to N-formylpirimicarb and, in a second step, to N-desmethylpirimicarb as well as to further oxygenated products with the carbamate moiety intact. Photolysis in cyclohexene resulted in photomineralisation as the main degradation pathway. Irradiation of pirimicarb in the presence of isopropanol also yielded an addition product of the parent compound with the secondary alcohol. PMID- 8237120 TI - [Child and adolescent psychiatry and Europe]. PMID- 8237121 TI - [Treatment of reading and writing disorders after the primary grades: application and evaluation of a concept]. AB - An empirical investigation is described which demonstrates the effectiveness of a program for dyslexic children (n = 50). The program was used with small groups of approximately 6 children within the school setting and continued for a period of 1 1/2 to 1 3/4 school years. The entire investigation lasted 5 years, so it was possible to repeat the treatment program in other schools, and additional schools participated by providing control groups. The criterion used to test for treatment effects was improvement in spelling on a standardized test after the participation in the program. The results showed highly significant improvement in writing and spelling both in the treatment groups and in the classes of which these groups were a part, whereas in the control groups there was no significant improvement between the first and second tests. An error analysis showed that the children in the treatment groups improved most in the areas that had been worked on in the program. PMID- 8237122 TI - [Drug treatment of 7- to 14-year-old children. Results of a parent survey]. AB - In comparison to other European countries, Germany has by far the highest availability of drugs. Consequently the fear is often expressed that drugs may be over-prescribed even for children and that educational measures or home remedies would often be equally effective. In particular, the issue of inappropriate prescribing of psychotropic drugs for children is often raised, even though epidemiological studies have failed to document such a practice. In a study conducted to obtain comprehensive data about the use of drugs by children, questionnaires were given to parents of children at four schools in Bavaria (2 industrial towns, 2 university towns). This anonymous inquiry was supported by the Ministry of Education and Cultural Affairs. Of the 1750 questionnaires distributed 1189 were evaluated (67.9% response rate). On an average, each child took 3.17 drugs per year, mainly for respiratory infections, fever, pain and gastrointestinal symptoms. Analgesics, nasal preparations, cough medicine, gastrointestinal medications and antibiotics were used frequently and psychotropic drugs only rarely (rank 20). For psychiatric conditions, only one child in 170 was given psychotropic drugs and only one in 108 used sedatives. Overall, drugs and home remedies were used with almost equal frequency (1.23:1). For psychiatric problems, home remedies were used three times more frequently, whereas for other problems drugs predominated. Of the parents who responded were 79.3% satisfied with the prescribed drugs; 9.1% stated that they changed the dosage of the prescribed drugs. PMID- 8237123 TI - [Phenylketonuria: illness experience and coping mechanisms]. AB - If patients with PKU follow a strict diet low in phenylalanine from soon after birth their intellectual and psychomotor development will be within the normal range. Recent studies have shown that it is necessary to continue the burdensome diet throughout one's life. This is in contrast to past practice and puts additional emphasis on psychosocial aspects of PKU. In the present retrospective study we investigated how adolescent patients and their relatives' experience and cope with this chronic disease. PMID- 8237124 TI - [Developmental psychopathology--the disease concept in child and adolescent psychiatry in comparison with the psychosomatic viewpoint]. PMID- 8237125 TI - [The adoption problem in treatment of a 15-year-old boy]. AB - In a brief review of the literature, the typical psychological problems associated with adoption are summarized. There is a marked discrepancy between the positive prognosis for adoption seen in the prospective studies on the one hand and the often dubious outcome of clinical treatment on the other. Our case study suggests that the apparently ubiquitous "issue of adoption" needs other co factors to become a "pathology of adoption". Clinical experience underlines the difficulty in and importance of finding an adequate therapeutic setting for severely disturbed adolescent adoptees: outpatient treatment is sometimes overtaxed by the tendency of the family to reject the adoptee and the noisy often dissocial behaviour of the adoptee, but an inpatient setting may repeat the trauma of separation, and often there is not a sufficient period of time for treatment. In this "therapeutic dilemma" where there is a high risk of personal relationships we see the need for a better connection between inpatient and outpatient treatment: The inpatient therapist should be able to continue treatment on an outpatient basis after the patient is discharged. PMID- 8237126 TI - [The need for quality standards in child and adolescent psychiatry]. PMID- 8237127 TI - Pharmacological properties of nicotinic acetylcholine receptors on adrenal chromaffin cells and their modification by hypoxic exposure. AB - To characterize the properties of nicotinic acetylcholine receptors (nAChRs) in autonomic ganglia, we examined specific L-[3H]nicotine binding to membrane fraction prepared from cultured bovine adrenal chromaffin cells, using a modified filtration method. Binding of L-[3H]nicotine to non-treated glass fiber filters interfered with detection of specific binding to the membrane fraction. Presoaking the glass fiber filters in 3% or greater concentrations of polyethyleneimine solution (sixty times higher than the reported concentration) for more than 5 h could reduce the binding of L-[3H]nicotine to the filters to background level. Furthermore, specific L-[3H]nicotine binding to the membrane fraction was detected only when the membrane fraction was prepared in buffer containing no divalent cations such as Ca2+ and Mg2+ (EDTA and EGTA were added) and protease inhibitors. Specific binding of L-[3H]nicotine was saturable and reversible. Scatchard analysis revealed a single class of high affinity binding sites with an average Kd of 8.7 nM and a Bmax of 43.2 fmol/mg protein. Specific binding was sensitive to cholinergic agonists (carbamylcholine and L-nicotine) and ganglionic stimulating agents (lobeline and DMPP), but was resistant to neuromuscular blocking agents (alpha-bungarotoxin, d-tubocurarine) and hexamethonium. These results suggest that high-affinity nicotine binding sites on adrenal chromaffin cells are nAChRs of ganglion-type, which have properties different from nAChRs on the neuromuscular junction but similar to nAChRs in the brain. PMID- 8237128 TI - The discriminative effects of nicotine and their central sites in rats. AB - The mediations in the central sites of the discriminative effects of nicotine were studied. Rats were trained to discriminate between the effects of nicotine at 0.5 mg/kg, sc and saline under a fixed ratio 10 schedule, for food reinforcement in a 2-lever operant chamber situation. After the establishment of discrimination behavior, guide cannulae were implanted bilaterally into the lateral ventricle (n = 10), the nucleus accumbens (n = 11), the ventral tegmental area (n = 7), and the dorsal hippocampus (n = 9). In substitution tests, nicotine at 100 and 120 micrograms per rat administered into the lateral ventricle and this drug at 100 micrograms per rat administered into the nucleus accumbens substituted for subcutaneous nicotine. On the other hand, neither nicotine at 10 80 micrograms per rat administered into the ventral tegmental area, nor this drug at 5-20 micrograms per rat administered into the dorsal hippocampus substituted for the subcutaneous nicotine. In antagonism tests, mecamylamine was administered into each brain area in combination with nicotine at 0.5 mg/kg, sc. Mecamylamine at 180 micrograms per rat administered into the nucleus accumbens blocked the discriminative effects of the subcutaneous nicotine while mecamylamine, administered into the ventral tegmental area and the dorsal hippocampus, did not. These results may indicate that the discriminative effects of subcutaneous nicotine are at least partially mediated in the nucleus accumbens. PMID- 8237129 TI - Effect of nicotine on cognitive disturbance in animals with acetylcholine deficient brain. AB - The effect of nicotine on learning disturbance was tested in rats fed with a choline-deficient diet (Ch free) or in senescence-accelerated mice (SAM). Concentration of acetylcholine in the brain was significantly lower in rats fed with Ch free and SAM. Passive avoidance learning shows that rats fed with Ch free and SAM showed significantly impaired learning, compared to controls. Nicotine (0.04 mg/kg) administered intraperitonealy significantly improved learning in animals with learning disturbances, as well as in controls. Vitamin B12 facilitated acetylcholine synthesis or release in the brain and improved the cognitive disturbance. We therefore suggest that acetylcholine contributes to learning and that nicotine may improve learning in an acetylcholine deprived rats. PMID- 8237130 TI - Age-related alterations in receptor density and metabolic activity in the brain. AB - The present experiments were designed to investigate whether or not age-related changes in brain function became detectable by comparing the responsiveness to stimulants or blocking agents between young and relatively aged rats. Oxygen consumption by hippocampal slices under the resting condition (3.5 mM KCl) was not significantly altered by aging (between 26 and 78 wk of age). When KCl concentration was increased to 50 mM, oxygen consumption by the slices from both young and aged groups significantly increased, but the increase was significantly greater in slices from young rats. Enzyme activities of the glycolytic pathway as well as a non-energy metabolizing enzyme, choline acetyltransferase, were not different between 6 and 26 wk of age. [3H]Prazosin binding in the cerebral cortex tended to be lower in the aged (78 wk old) rats but not significantly different from that in the young (13 wk old) ones. On the other hand, the recovery from the reduced prazosin binding following administration of phenoxybenzamine was significantly delayed in the aged rats. These results indicate that the age related changes in brain function become evident by comparing the responsiveness to stimulants or blocking agents between young and aged rats. PMID- 8237131 TI - A study on cerebral nicotine receptor distribution, blood flow, oxygen consumption, and other metabolic activities--a study on the effects of smoking on carotid and cerebral artery blood flow. AB - We investigated middle cerebral artery flow velocity (MCA-FV) by a noninvasive method to determine whether or not smoking causes an increase in cerebral blood flow (CBF). Furthermore we determined sequentially the changes in CBF caused by smoking in order to evaluate changes in responses at different times in daily activities and the effect from meals. The subjects were 25 healthy individuals ranging in age from 20 to 36 yr. MCA-FV was measured by a transcranial Doppler system. They smoked a filtered cigarette for 5 min at 1 P.M., 3 P.M., 6 P.M., 8 P.M., 10 P.M., 8 A.M., and 11 A.M. Results (1) Smoking caused increases in both common carotid artery flow volume and MCA-FV, and the percentage increase of these parameters showed a good correlation (r = 0.809). (2) MCA-FV increased significantly during the first (by 6.6%) and second halves (by 5.4%) of the smoking period. (3) The change in MCA-FV after meals was slight. (4) Smoking tended to increase MCA-FV during each smoking session but the changes were not significant. The pulsatility index reduced significantly during almost every smoking session. These results lead to the conclusion that smoking reduces vascular resistance in cerebral arteries and increases CBF. PMID- 8237132 TI - Studies on the effects of smoking on cerebral perfusion and metabolism: basic studies with in vivo tracer methods. AB - In order to examine the effects of smoking on the central nervous system, we applied the radionuclide method to study the in vivo behavior of nicotine and the effects of nicotine on cerebral perfusion and metabolism. Biodistribution studies with optical isomers of carbon-11 labeled nicotine showed greater uptake and prolonged retention of (S)-nicotine in the brain with competitive inhibition by cold nicotine, suggesting the possibility of functional assessment and imaging of nicotinic receptors in human brain. Senescence accelerated model mouse, which was reported to show age related deterioration of memory, demonstrated decreased accumulation of nicotine in the brain. Intravenous injection of nicotine revealed markedly increased cerebral glucose metabolism with regional differences, while it slightly reduced cerebral blood flow. These results suggest that in vivo studies with radiolabeled tracers will play a significant role not only in clarifying the effects of smoking or nicotine but also in understanding the mechanism of interaction between neuronal function and cerebral perfusion and metabolism. PMID- 8237133 TI - Distribution of 11C-(R)nicotine in human brain measured by positron emission tomography: measurement of cerebral blood flow. AB - We synthesized 11C-labeled (R)nicotine (11C: a positron-emitting radioisotope with a half-life of 20 min, produced in a cyclotron), and utilized it for blood flow measurement in the human brain. The positron emission tomography was employed for the data acquisition and image reconstruction. The behavior of the isotope was formulated by a mathematical model, consisting of a blood compartment and a tissue compartment, and the time-activity curve in various parts of the brain was analyzed in a computer to obtain a mathematically simulated curve which fell in consistence with both the arterial blood curve and the brain curve. The cerebral blood flow (CBF) value thus determined was compared with the value obtained by the standard C15O2 inhalation method (= H2(15)O method). The nicotine method always gave a higher value than the standard method by 27-34%. It seems that CBF determined by the 11C-(R)nicotine method is more accurate than the value determined by the standard method, because our tracer is superior as a CBF indicator to radiolabeled water. PMID- 8237134 TI - Non-invasive assessment of acute effects of cigarette smoking on cerebral circulation. AB - The usefulness of sequential single photon emission computed tomograph technique using Tc-99m hexamethylpropyleneamine oxime was tested to assess the acute effects on cerebral circulation of cigarette smoking and other activation task. The evaluation of reproducibility, photo-stimulation study, and carbon dioxide inhalation study were carried out. In the subjects, who were examined for cerebral perfusion under the resting condition repeatedly, no significant perfusion changes were detected in visual analysis, in quantitative analysis, or in distribution change mapping. The significant increase of CBF in the occipital lobes was found by photo-stimulation. The CO2 reactivity, estimated by a pair of perfusion images obtained under normocapnia and hypercapnia, was lower in the affected side than in the non-affected side of the patients with unilateral major occlusive lesions in the carotid system. These results showed our method was applicable to assess acute effects on regional CBF of cigarette smoking. Five normal volunteers were investigated for the CBF response to cigarette smoking. Although localized CBF increases were detected in two cases, no characteristic pattern could be found. These results suggested that in normal subjects, the effects of cigarette smoking on regional cerebral circulation varied among individuals, depending on the pharmacological properties of the absorbed nicotine including different sensitivity of each subject to nicotine. PMID- 8237135 TI - Effect of chronic smoking on regional cerebral blood flow in asymptomatic individuals. AB - Correlations between cigarette smoking and cerebral blood flow (CBF) were determined. The purpose of the present study was to determine whether the level of cigarette use is a significant predictor of regional CBF (rCBF) when age, gender, mean arterial blood pressure, total cholesterol, fasting blood glucose, hematocrit, and presence of ST-T change and left ventricular hypertrophy on electrocardiogram (ECG) are controlled. We studied a continuous sample of 40 asymptomatic individuals including 20 smokers and 20 nonsmokers. Subjects (mean age 66.2 yr) had an average smoking index of 456 +/- 485.1 (mean +/- standard deviation) [(Number of cigarette/d) x (years of smoking history)]. Regional CBF was measured using the intravenous 133Xe injection method. Simple linear regression and multivariate regression analyses were performed, which modeled regional cerebral blood flow as a function of smoking index and other cerebrovascular disease risk factors. The male-to-female ratio was higher in the group of smokers (18/2) than in the group of nonsmokers (2/18) (P < 0.01). The mean hematocrit of smokers was significantly higher than that of nonsmokers (P < 0.01). There were no significant differences in other variables tested between the two groups. Simple linear regression analysis demonstrated a significant negative correlation of smoking index with CBF in whole brain (r = -0.33; P < 0.05), the right hemisphere (r = -0.34; P < 0.05), right parietal cortex (r = 0.36; P < 0.05), right occipital cortex (r = -0.34; P < 0.05) and left parietal cortex (r = -0.33; P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237136 TI - Review on the relationship between nicotinic acetylcholine receptors and dopaminergic neurotransmission in the central nervous system--dopa is an endogenous neuroactive substance. AB - L-3,4-Dihydroxyphenylalanine (DOPA) is believed to be an inert precursor for dopamine (DA). Contrary, transmitter-like endogenous DOPA is released from in vitro and in vivo striata: DOPA is released by neuronal activities under physiological conditions from striata of conscious rats. Furthermore, exogenous nanomolar DOPA itself produces an in vitro presynaptic response to facilitate the catecholamine release. An in vivo postsynaptic depressor response is elicited by DOPA microinjected into the nucleus tractus solitarii. These responses are antagonized by L-DOPA methyl ester, a competitive DOPA antagonist. In striata, DOPA is an endogenous potentiator for presynaptic beta-adrenoceptors to facilitate the DA release and also probably for postsynaptic D2-receptors to increase locomotor activities. Nicotine releases DA and transmitter-like DOPA in vitro and in vivo striata. Nicotine (0.1-1.0 mg/kg, sc) dose-dependently increases locomotor activities. This increase is stereoselective and mecamylamine (1.0 mg/kg, sc)-sensitive but not antagonized by L-DOPA methyl ester (200 micrograms, ivt). Then, a selective low ip dose of alpha-methyl-p-tyrosine (alpha MPT) to inhibit the basal release of DOPA without decreasing the basal release of DA was explored in vivo striata: it was 3 mg/kg. Pretreatment with this dose did inhibit the nicotine-induced increases in locomotor activities. This result suggests that endogenously released DOPA is in part relevant to nicotine-induced behavior in rats. PMID- 8237137 TI - Impairment of learning and memory and the accessory symptom in aged rat as senile dementia model (3): Oral administration of propentofylline produces recovery of reduced NGF content in the brain of aged rats. AB - The relationship between nerve growth factor (NGF) and senile dementia of the Alzheimer type is of interest. The administration of NGF in clinical trials has been expected; however, NGF cannot cross the blood brain barrier, since it is a polypeptide. We demonstrate here that oral administration of propentofylline, a stimulator of NGF synthesis in vitro, produced recovery of reduced NGF content. In aged rats (24-months-old), NGF content in the frontal cortex, parietal cortex, and hippocampus was lower than in young rats (8-weeks-old). Twenty-one-day successive administration of propentofylline (10 and 25 mg/kg, p.o.) produced recovery of reduced NGF content in the frontal cortex and parietal cortex of aged rats to a significant extent. There were no differences among aged, young, and propentofylline-treated groups in choline acetyltransferase activity in any brain regions. These results suggest that NGF content in the brain is reduced in aged rats and that oral administration of propentofylline leads to a recovery of this reduction. PMID- 8237138 TI - [Therapeutic and adverse effects of zotepine and their relationships with serum kinetics of the drug]. AB - The spectrum of therapeutic and adverse effects of zotepine and their relationships with serum concentrations of the drug were investigated in 28 schizophrenic inpatients by a fixed-dose (100 mg/day for the first 1 wk and 200 mg/day for the following 3 wk) study. The mean % improvement in Brief Psychiatric Rating Scale (BPRS) scores were 63.4% for total, 64.4% for positive, 41.8% for negative and 74.2% for anxiety-depression symptoms, suggesting that zotepine has a broad therapeutic spectrum. There were significant differences in improvement of negative symptoms after 1 wk and total and positive symptoms after 2 wk (P < 0.05) between responders (more than 50% reduction in total BPRS scores at wk 4) and nonresponders. The mean scores of UKU Side Effect Rating Scale were low in this study. UKU scores of psychic at wk 2 and total adverse effects at wk 3 and wk 4 (P < 0.05) were higher in nonresponders than in responders. A negative correlation between % improvement in total BPRS and total UKU scores was observed (P < 0.05). These two results suggest some relationship between poor clinical response and increased adverse effects during zotepine treatment. Lack of close correlation between serum drug concentrations and clinical or adverse effects indicates that therapeutic monitoring of zotepine concentrations would be of little significance. PMID- 8237139 TI - [A controlled study of the prophylactic effect of mianserin and placebo in recurrent depression]. AB - To examine the prophylactic effect of mianserin on depressive episode, 26 patients with recurrent depression were randomly allocated on a double-blind basis to treatment with either inactive placebo (PL group) or mianserin in daily dose of 20-60 mg (MIA group) for 18 months and the frequencies or durations of depressive episode during the treatment were compared between the two groups. Except for the 4 patients who dropped out, a comparison of the prophylactic effects of the two groups was performed on 9 patients of the MIA group and 13 patients of the PL group. As a result, recurrence of depressive episode occurred in 4 of 9 patients in the MIA group and in all 13 patients in the PL group during the study. The recurrence rate of the depressive episode during the period in the MIA group was always less than that in the PL group, and significant differences were observed between the two groups from the third to the 18th month of the study. Whereas the frequency and duration of the depressive episode in the MIA group during the period of the study were significantly less than those prior to the study, there was no such difference in the PL group. On the other hand, there was no significant difference in the safety between the two groups. From the result of this study, it was suggested that mianserin is effective in the prophylaxis of depressive episode in patients with recurrent depression. PMID- 8237140 TI - Cholinergic modulation of spatial working memory of mice in radial maze performance: retention curve analysis. AB - Cholinergic modulation of the spatial working memory of mice was investigated in an eight-arm radial maze. The mice were trained to achieve a stable baseline level of performance, and the retention of spatial working memory was then examined by means of imposition of retention intervals of 45, 90, 180, and 360 min between choices four and five. The animals were removed from the apparatus during these intervals. The effects of oxotremorine (0.025-0.1 mg/kg, ip), physostigmine (0.025-0.1 mg/kg, ip), and scopolamine (0.1-0.4 mg/kg, ip) on the number of the correct responses after the various time intervals (5-8 choices) were studied. Mice received drug injections 30 min prior to the first four arm choices before delay intervals (1-4 choices). The number of correct responses after the intervals decreased with the increased length of the retention interval. Oxotremorine and physostigmine exhibited dose-dependent stimulating effects of resistance to decreases in the number of correct responses, but scopolamine potentiated the decline in a dose-dependent manner. Significant interactions between the effects of cholinergic drugs and the length of retention over time were also observed. These observations indicate a cholinergic modulation of spatial working memory in mice engaged in radial maze performance, as assessed by means of retention curve analysis. PMID- 8237141 TI - [Clinical significance of plasma levels of clomipramine and its desmethylated and hydroxylated metabolites]. AB - Concentrations of clomipramine and its metabolites, N-desmethylclomipramine (DC), 8-hydroxy-N-desmethylclomipramine (HDC) and 8-hydroxyclomipramine (HC), in plasma were determined in 99 patients treated with clomipramine hydrochloride. Doses patients received were not fixed but titrated according to their clinical severity and response to the treatment. Large interindividual variations were present in the concentrations of parent or each metabolic compounds in plasma, however, strong correlations existed between these drug concentrations and daily doses of clomipramine (0.40-5.10 mg/kg of body weight) (r = 0.62-0.80). Metabolic ratios for desmethylation and those for hydroxylation were calculated from these data, and revealed very large inter-individual variations of 30-36 fold and 10-96 fold, respectively. Pharmacological data from 65 of these patients with DSM-III-R mood disorders were analyzed by discriminant analysis using the scores of Global Assessment of Functioning. The overall prediction value for responder or nonresponder was calculated as 70% 2 wk after the initiation of pharmacotherapy. Concentrations of hydroxylated metabolites in plasma does contribute as much to the prediction of clinical response to clomipramine as desmethylated metabolites. PMID- 8237142 TI - [Molecular genetics of aromatic L-amino acid decarboxylase]. AB - Aromatic L-amino acid decarboxylase (AADC) decarboxylates both L-5 hydroxytryptophan to serotonin in serotonergic neurons and pineal cells, and L dopa to dopamine in catecholaminergic neurons and adrenal medullary cells. Thus AADC produces two major mammalian neurotransmitters and hormones. We isolated and sequenced a full-length, neuronal-type, cDNA encoding human AADC. It consisted of 1932 bases containing an open reading frame encoding 480 amino acids residues with a molecular weight of 53,891. We expressed a recombinant human AADC in COS cells and proved that the expressed enzyme decarboxylated both L-5 hydroxytryptophan to serotonin and L-dopa to dopamine. We have cloned genomic DNA of human AADC and determined the structure. The genomic DNA of human AADC consists of 15 exons spanning about 100 kilobases and exists as a single copy in the hapoloid genome. We have mapped the gene to chromosome band 7p12.1-p12.3 by fluorescence in situ hybridization. We cloned the nonneuronal type cDNA from human liver and identified another first exon different from the neuronal type cDNA. This showed that an alternative usage of the first exon produced two types of mRNAs in AADC and suggested that alternative splicing would regulate the tissue-specific expression of AADC. PMID- 8237143 TI - [Opioid receptors in the brain related to behavior]. AB - The effects of opioid peptides selective for opioid receptors on drinking, dopamine-related behavior, discriminative stimulus and memory processes were investigated in detail. Mu opioid receptors in the paraventricular nucleus of hypothalamus played an important role in drinking behavior. The stimulation of mu and kappa opioid receptors inhibited dopamine D2 receptor-related behavior. The discriminative stimulus effects of morphine and pentazocine were mediated through mu opioid receptors, while the stimulus effects of cocaine through delta opioid receptors. Mu opioid agonists elicited memory dysfunctions, whereas kappa opioid agonists such as dynorphin A-(1-13) improved amnesia induced by ischemia, basal forebrain-lesion or scopolamine. It appears that opioid receptors are fully involved in the neural basis of behavioral responses. PMID- 8237144 TI - [Senile dementia in institutions for the elderly--agreement rate between clinical and pathological diagnoses]. AB - A study was performed to assess the occurrence of senile dementia in institutions for the elderly. In a total of 747 cases examined in 1986 we diagnosed 316 (42.3%) to be suffering from dementia. The patients were subdivided by etiology into 4 categories: Vascular type (VD) 51.6%, Alzheimer type (DAT) 21.8%, Mixed type 9.2% and others 17.4%. Subsequently 124 of these demented patients died and autopsies were performed on 104 cases by 1990. Final clinical diagnoses were made and correction of initial diagnoses was necessary in 15 cases. The agreement rate between final clinical and pathological diagnoses was 79.8% for all 104 cases where data was available. For individual categories treated separately, the rates were VD 83.3%, DAT 89.3%, Mixed 64.7% and others 63.6%. PMID- 8237145 TI - Studies on the refrigerated storage of wheat (Triticum aestivum). 2. Ergosterol, xanthomegnin, viomellein and brevianamide A after inoculation with Penicillium viridicatum. AB - Wheat seed was adjusted to 18, 20, 22, 24 and 26% moisture content (m.c.), and stored for 240 days at 4 or 10 degrees C following inoculation with a strain of Penicillium viridicatum producing the toxins, xanthomegnin (XA), viomellein (VIO), and brevianamide A (BA). Wheat kernels were not sterilized before inoculation. The concentration of ergosterol (ERG), a chemical indicator of fungal biomass, remained constant at 18% m.c./4 degrees C, but increased under the other conditions. The time before a detectable increase of ERG concentration was higher and the rate of ERG production lower with decreasing moisture content and temperature. XA and BA were produced at both temperatures at 20-26% m.c., VIO was produced at 22-26% m.c./4 degrees C and 20-26% m.c./10 degrees C. The results suggest or indicate that the onset of XA, VIO and BA production (detection limits: 10, 15, and 0.1 micrograms/kg, respectively) coincided with the onset of ERG production. Maximum toxin contents were lower with decreasing moisture content at both temperatures, but were similar at 4 and 10 degrees C at 22-26% m.c. It is concluded that wheat contaminated with P. viridicatum should not be stored beyond the onset of ergosterol production; maximum storage periods are recommended. PMID- 8237146 TI - [Importance of tumor growth and spread for surgical radical excision]. AB - Different organ tumours of the gastrointestinal tract show variations in invasion patterns which are partly dependent on the biological characteristics of the tumour and partly dependent on the anatomical situation of the involved organ. It is important for the surgeon to have knowledge of the local invasion behaviours in making a precise indication for surgery, in the planning and in the technical accomplishment of the operation. Residual tumour has been shown to be a particular important prognostic factor. This paper investigates a large series of patients with different organ tumours in order to find in which location a R1 situation must be expected and which surgical consequences should be thus derived. The percentage of R0 resections in all patients who underwent resection differed in the individually examined organs: oesophagus, 65%; stomach, 80%; pancreas, 77%; colon, 81%; rectum, 84%. R1 resections also showed differences in incidence: oesophagus, 17%; stomach, 7%; pancreas, 13%; colon, 3%; rectum, 4%. Residual tumour was most frequently found in the lateral (superficial or free) resection margins: oesophagus, 79%; stomach, 67%; pancreas, 86%; colon, 78%; rectum, 74%. The other resection margins were significantly less-frequently involved. PMID- 8237147 TI - [Is surgical therapy of locoregional recurrence of resected colorectal cancer effective?]. AB - The role of surgery in the treatment of local recurrence of colorectal carcinoma is controversial. This study aims to analyse the value of surgical reintervention. From July 1982 to June 1991 157 patients were treated for local recurrence after "curative resection" of primary colorectal carcinoma in our institution. The mean age was 60.3 years. In 67 patients tumor recurred in the pelvis (45% distant metastases) and in 90 patients in the abdomen (39% distant metastases). Intraluminal recurrence was found in 55 patients (35%), extraluminal recurrence in 102 patients (65%). The mean interval between primary tumor resection and recurrence was 24 months. The resection rate was 73.9% (n = 116). R0-resection was achieved in 38% (n = 44). There was no significant difference in the R0-resection rate between pelvic and abdominal recurrences (33% vs. 41%). In contrast to that intraluminal tumors showed a significant difference in R0 resections when compared with extraluminal tumor recurrences (57% vs. 23%). In 34 patients (22%) only palliative surgery was performed, while in 7 patients (45%) no surgical intervention was possible at all. Multivisceral resections were necessary in 36% of pelvic and in 81% of abdominal recurrences. The postoperative complication rate was 58% (19% major complications), mortality was 3.4%. For R0 resection mean survival was 44.1 months, for R1-resection 16.5 months and for R2 resection 10.9 months. Survival in intraluminal recurrences was 42 months compared with 13.8 months in extraluminal tumors. Tumor location (abdominal/pelvic) had no influence on survival.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237148 TI - [Prognostic factors in breast cancer--classification in non-university hospitals]. AB - Prognostic factors in breast cancer have been frequently discussed in the oncologic literature. We reviewed our data of 795 patients who were referred to the Department of Radiation Oncology of the University of Wuerzburg between 1978 and 1988 in order to find out, whether the well known and proven prognostic factors in breast cancer were sufficiently in use in clinical routine, even out of university oncologic centres. We found, that beside of determination of the histological tumor-type the assessment of tumor size (= pT-status) and axillary (= pN-)status is performed nowadays in all patients. In contrary, the determination of the hormonal receptor status (HRS) had a slow beginning. From 1980 the rate of determinated HRS arose from 5% to 80% nowadays. The demand of removal and examination of at least 12 axillary lymph nodes in axillary dissection in order to determine correctly the axillary status is not yet part of the oncological routine. In 1988 in only 40% of the referred patients axillary diagnosis was done in a manner corresponding to international rules. PMID- 8237149 TI - [Surgical treatment strategies in femoral metastases]. AB - Because of the increasing survival-time of tumor-patients bone metastasis is becoming an important subject in surgery. In the case of osteolysis of the femur a concept of treatment in consideration of various localisations is presented. In 81 patients with pathological fractures of the femur or osteolysis in this region from January 1987 to June 1992 96 stabilizing operations were performed. Almost one half of the patients suffered from breast carcinoma followed by hypernephroma, Plasmocytoma, bronchial-carcinoma and prostate-carcinoma. The average survival time after stabilization of the femur was 8.3 months. After the operation the condition of the patients improved on an average by 0.64 units of the modificated Karnowsky-Index. Surgical treatment of osteolysis in the femur is indicated either in pathological fractures or if instability is imminent. Moreover patients who suffer from severe pain should also receive palliative surgery. With regard to the site of the lesion various types of endoprostheses and osteosynthesic devices can be used. If the femoral neck or the intertrochanteric region is affected arthroplasty with cemented endoprosthesis may be performed. In subtrochanteric osteolysis long and thin endoprosthesis can be used as well as Gamma-nails. Pathological fractures in the shaft of the femur should be treated with intramedullary static locked nails, which can be strengthened by supplementary lateral plates. If the distal metaphysis of the femur is involved compound osteosynthesis with methylmethacrylate and dynamic condylar screws or angle plates can be used. PMID- 8237150 TI - [Extended resections in bronchial cancer: complications and late results]. AB - From 1982 through April 1992, 192 patients underwent extended resection for non small cell carcinoma of the lung (NSCLC) invading the chest wall (n = 52) or the mediastinum (n = 140). Complete resection of the tumor was possible in 81 (58%) patients with invasion of mediastinum and in 34 (65%) patients with invasion of the chest wall. The operative mortality was 9%, respiratory complications causing most of the postoperative morbidity and mortality. Incomplete resection was associated with poor prognosis. The mean survival of these patients was 10 month. The 5 year actuarial survival rate of patients having curative resection was 20%. In these patients the presence of lymphatic metastases significantly reduced survival, with a 3 year actuarial survival rate of 40% of patients with No disease, and 30% or 5% for those with N1 or N2 disease, respectively. In conclusion, extended resection for advanced NSCLC offers a significant chance for long-term survival in the absence of N2 lymphatic metastases. In the presence of N2 metastases and poor overall prognosis however, reasonable palliation is the primary goal of surgical management. PMID- 8237151 TI - [Expandable metal mesh stents for treatment of tracheal stenoses and tracheomalacia]. AB - The treatment of tracheo-bronchial stenosis or tracheomalacia is mainly carried out by means of resection or tracheoplastic operative strategies. Since the introduction of metal-mesh stents, a definitive endoluminal therapy has to be considered under new aspects. Six patients with malignant stenosis or tracheomalacia due to compression were treated by implantation of Palmaz- or Wallstents. Immediately after the implantation, patients were relieved from dyspnoea, the forced inspiratory volume-1 (FIV1) was normalized. All implanted stents were well tolerated, even in the long-time follow-up (19 months). Bronchoscopic control showed overgrowth of the metal meshes by respiratory epithelium. The implantation of metal-mesh stents is an adequate alternative in the treatment of malignant stenosis and tracheomalacia. PMID- 8237152 TI - [Video-thoracoscopic atypical lung resection]. AB - A new era in thoracoscopy within the framework of minimal invasive thoracic surgery was introduced due to the possibilities of video-endoscopy, the development of an accordingly advanced and suitable instrumentarium as well as the production of an endoscopically applicable linear stapler. The introduction of the endo-staplers has made the thoracoscopic resection even of larger parts of lung parenchyme possible. Just as in laparoscopy a clear reduction of post operative pain, a reduced impairment of post-operative pulmonary function, a shorter in hospital stay and an earlier return to normal activities are advantages of thoracoscopy when compared to the severely more traumatising thoracotomy. Good indications for this procedure are the spontaneous pneumothorax and the resection biopsy in the quest of finding the diagnosis of unclear pulmonary disease. For the diagnostic and should the situation arise therapeutic resection of solitary pulmonary nodules in contrast a stricter selection and handling of indications is imperative. The method however proved worthwhile under such conditions. The technique and results are reported of video thoracoscopies in 73 patients as well as 51 atypical lung resections carried out thoracoscopically with the Endo-GIA 30. A switch to open thoracotomy became necessary in 16.4%. Indications were pneumothorax (35.6%), hemothorax (1.3%), diagnostic resection of solitary nodule (42.5%), pneumopathy (11.0%), and extrapulmonary tumor (9.6%). Atypical resections were carried out only in the case of solitary nodule (51.7%), pneumothorax (39.7%), and pneumopathy (8.6%). A histological diagnosis was obtained in all cases. The average operating time for solitary nodule and pneumothorax was 1 hour, for pneumopathy 1/2 hour. Chest drains could be removed after 1.6-2.6 days. The postoperative complication rate amounted to 3.9%. There was no mortality. The introduced procedure represents a new and promising alternative in thoracic surgery. PMID- 8237153 TI - [Use of TNM classification in clinical practice]. AB - We have retrospectively evaluated the use of TNM in patients referred to our clinic for radiation treatment between 1979 and 1990. The percentage of TX-cases (unknown and not exactly classified primaries) decreased from 22% in 1979 to 3% in 1990 and was lowest in tumors treated according to well-defined treatment protocols (e.g. head and neck tumors, breast cancer). PMID- 8237154 TI - [Current focus: blood transfusion. Risks and problems in blood transfusion]. AB - Blood transfusion became the subject of major interest because of a possible HIV infection. It should not been forgotten, however, that HIV-infection is not the only of a wide variety of risks. The leading complication is still the hemolytic transfusion reaction, followed by transmission of hepatitis or other viral or bacterial germs. Less striking side effects, like an immunomodulation or a graft versus-host reaction, should also been included into the therapeutical considerations. One possibility to avoid risks, is the use of autologous blood. The most important requirement, however, remains to operate with a minimal blood loss. PMID- 8237155 TI - [Blood transfusion 100 years ago]. PMID- 8237156 TI - ["The interesting case" (issue 6 (1993) 368-71). T. Zimmermann, C. Kelm, C. Heirichs, G. Herold: Bilateral quadriceps tendon rupture]. PMID- 8237157 TI - [Endothelin--possible significance in pregnancy and hypertensive pregnancy]. AB - Endothelin (ET) is the most potent endogenous vasoconstrictory substance known. There are three structurally and pharmacologically separate endothelial isopeptides in humans; Endothelin-1 is exclusively produced in the vascular endothelium. It seems likely that ET acts as a local paracrine signal rather than a circulating hormone. The synthesis and release of ET is stimulated among others by hypoxia, thrombin and endotoxin. Its effects are mediated by specific, membrane-bound receptors, which are detectable in high concentrations in the fetoplacental tissue. ET-1 causes an initial transient fall in blood pressure, followed by a strong, long-lasting increase in peripheral resistance and blood pressure. Plasma ET-1 levels are increased in preeclampsia as compared to those of normal pregnancies, and do not correlate with mean arterial blood pressure and degree of proteinuria. In umbilical cord blood ET-1 concentrations are 2.5-10 fold higher than those of maternal plasma. Determination of plasma ET is unlikely to be of value in the prediction of the disease. ET-1 induces an increased synthesis of vasodilatory prostaglandins (PGI2, PGE2) and an increased production of endothelial-derived relaxing factor (EDRF); thromboxane concentrations in blood are elevated by thrombin-induced activation of platelets. In animal models ET-1 causes an activation of plasmatic coagulation with consecutive hypercoagulability. In preeclampsia ET may play an important role in the regulation of the endothelial balance. Future therapeutic approaches may include the development of effective ET-antibodies or of inhibitors of the endothelin converting enzyme. PMID- 8237158 TI - [Intervals for increasing the dosage of oxytocin]. AB - We retrospectively analyzed 468 deliveries in 1989, 935 deliveries in 1990, and 1020 deliveries in 1991 from cephalic presentation. Oxytocin was given for reason of not sufficient labor (cervical dilatation < 1 cm/hr) after spontaneous or artificial rupture of membranes. Oxytocin was increased at intervals of 20 minutes (in 1989) or 60 minutes (in 1990 and 1991). The percentage of deliveries augmented with oxytocin, the cesarean section rate in deliveries with or without oxytocin, the maximum oxytocin dose, and the condition of the neonates (arterial cord blood pH value, Apgar scores at 1 and 5 minutes) were compared. Prolonging the interval of increasing oxytocin did not adversely influence the condition of the neonates and was not associated with a significant change in the cesarean section rate. The average duration of oxytocin administration was prolonged slightly, but the maximum dose and therefore the average total dose administered were decreased. PMID- 8237159 TI - [Laser therapy of the vulva, perineal area and uterine cervix]. AB - Within a period of two years 205 patients of our colposcopy-laser-outpatient department have been treated with a 40 Watt CO2-laser. 97 patients had dysplastic or condylomatous lesions of the portio, 85 ones condylomas in the vulvar, vaginal or perianal regions. 70 per cent of laser vaporisations had been done ambulatoryly. Complication rate following laser conisation was 12 per cent, following laser vaporisation 0 per cent. The rate of recurrent or persistent dysplasias was 12 per cent, of condylomas 18 per cent on an average observation time of 16 months. PMID- 8237160 TI - [The value of Doppler imaging of the aorta, carotid artery and middle cerebral artery in obstetric diagnostics]. AB - The relation of blood blow in the fetal aorta and carotis is a marker for the well being of the unborn child. Increase of cerebral vascularity associated with decrease of blood flow in the aorta is a sign for a fetal risk situation. It is, however, of disadvantage that the fetal carotis is often difficult to image with an wedge-shaped angle between doppler beam and vessel. We therefore included screening of an intracerebral vessel (a. cerebri media) in order to find out whether examination of this vessel, much more easier to perform, may have a similar diagnostic value or be an even more sensitive prognosticator of fetal risk than the carotis. In two hundred and forty patients in the 29th to 40th week of gestation all these three vessels were scanned with the duplex system AI 3200, Dornier company, Germany. The result was correlated to sonographic biometry, birth weight, and mode of delivery. It could be proven that when analyzing individual findings, pathology in the risk groups is much higher, whereas mean values showed less marked differences between risk and control groups. Our results suggest that the a. cerebri media is a slightly more sensitive indicator of fetal risk, since in this vessel retrograde resistance can be detected earlier and is more evident. A combination between assessment of the aorta and a cerebral vessel, is however, the method of choice in cases of pathological findings in diagnosis of fetal well being, since only examination of a peripheral and a central vessel results in an exact diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237161 TI - [Breast asymmetry--differential diagnosis and surgical correction]. AB - The problems of asymmetry of the female breast have been exemplified on four cases. Changes in a single breast due to anatomical defects or as a result of an accident, as well as changes due to a large fibroadenoma have been presented. Surgical correction was made after an individual evaluation of each case and included reduction mammoplasty, breast augmentation, lifting and nipple transplantation or reconstruction. PMID- 8237162 TI - [Clinical aspects of primary malignant melanoma of the vagina]. AB - This is a German case report of 12 patients with malignant melanoma of the vagina. Their median age is 58.3 years. Four out of 12 patients have been treated by radical surgery alone. In 8 patients surgery was restricted to a wide excision. In four cases additional brachytherapy was performed. The prognosis of malignant melanoma of the vagina is extremely poor. Three patients survived for two years following primary therapy. All patients died within 6.5 years after diagnosis and initial treatment. In our experience and according to literature radical surgery seemingly achieves the best results within the limited prognosis of this disease. PMID- 8237163 TI - [Primary intracerebral non-Hodgkin's lymphoma--a clinicopathologic study]. AB - Primary cerebral lymphomas (PCL) were diagnosed with increasing frequency also in our retrospective study of 44 patients. Clinically these tumors presented with signs of a rapidly growing brain neoplasm. The analysis of CCT data showed that the tumors were of varying density before and showed mostly (60%) homogeneous enhancement after contrast medium application. MR imaging was more sensitive, but could not aid in distinguishing PCL from other brain tumors. The lesions lay mainly (82%) in the supratentorial space and involved the frontal lobe in 42% of cases. Only 16% were located in the periventricular region including corpus callosum and basal ganglia. 20% of cases showed multiple lesions. Suspected diagnoses were therefore mainly meningeoma, glioma and metastases. Morphological diagnosis was easily possible with the aid of immunohistological methods: there were 41 B-cell lymphomas (93%), two T-cell lymphomas and one large cell anaplastic lymphoma of the non-B non-T phenotype. An unequivocal correlation between morphology and radiological picture existed in the way that tumors with a dense cellular infiltrate appeared mainly as hyperdense lesions with homogeneous contrast enhancement. The clinical course was characterized by CNS-relapses frequently with multiple cerebral lesions and a spinal recurrence in one case. 7% of cases showed evidence of extracerebral disease in a bone marrow biopsy specimen or at autopsy. The average survival of the patients was 15 months, one year survival was 36%, two year survival 12% and 5% of patients lived for more than 5 years. PMID- 8237164 TI - [Tuberculum sellae meningioma and hypophyseal adenoma in a woman]. AB - The case of a 61 years old female patient with coincidental tuberculum sellae meningeoma and pituitary adenoma is demonstrated. Because of deteriorating vision of the left eye and bitemporal hemianopia the patient was investigated. Primary arranged computertomographical investigation showed a tall intra- and suprasellar solid homogeneous tumor. The plain skull film revealed sellar enlargement; angiographically craniolateral displacement of Aa. cerebri anteriores was found. Only MRI cleared up two coexisting tumors, an intrasellar lower enhancing tumor close to another extra-/parasellar. Neuroendocrinological investigations including insulinhypoglycaemiatest and the combined pituitary anterior lobe test showed findings to be seen typically in tuberculum sellae meningeomas. Only very discrete signs for a disturbance of anterior lobe secretion could be found. Postoperatively only a discrete deterioration of clinical symptoms and neuroendocrinological findings were noticed. The diagnosis of two coincidental tumors was confirmed histologically. Two years after operation no tumor recurrence was seen. This case is comparable with cases published until now. PMID- 8237165 TI - "Inverted sella turcica" appearance on skull X-ray due to chronic sphenoidal sinusitis. A report of two cases. AB - This paper presents two cases with a peculiar shape of the sella turcica on skull X-ray due to chronic sphenoidal sinusitis. This unusual "inverted sella turcica" sign, not previously reported, may be specific to this disease. PMID- 8237166 TI - Transsphenoidal extradural chiasmapexy in the management of the symptomatic primary empty sella syndrome. AB - When patients with the usually clinically silent primary empty sella syndrome become symptomatic through visual field defects or hormonal deficits the condition was frequently treated by a transsphenoidal intradural procedure with a risk of consecutive CSF leak or meningitis. We present 3 cases in which we performed a transsphenoidal extradural procedure packing the sella and sphenoid sinus with fat tissue and discuss the procedure and results. PMID- 8237167 TI - [Intracranial chordoma. Case report of a destructively growing chondroid chordoma in the area of the sella turcica]. AB - Evaluating the literature a short survey is given about chordomas of intracranial localization. A case of a chondroid chordoma growing destructively in the base of the skull is described. Because of the localisation in the region of the sella turcica the first diagnosis was an adenoma of the hypophysis or craniopharyngioma. Although it is a chondroid chordoma 14 months after operation a recidive tumour was seen. PMID- 8237168 TI - Estrogen and progesterone receptors in meningiomas and gliomas. AB - Tumor biopsy specimens from 15 patients with meningiomas and 15 patients with malignant gliomas were examined for estrogen and progesterone receptors. The frequency of receptor positivity is presented and results discussed along with the relevant literature. PMID- 8237169 TI - Metals in tumors of the sellar region. AB - The contents of the metals Ca, Mg, Zn, Cu, Cr, and Mn in 7 different entities of tumors of the sellar region were evaluated by atomic absorption spectrophotometry. Increased values of Mg in coincidence with high Ca contents are explained by the competition of Mg to Ca and its potency to inhibit mineralization. Reports on the frequent coincidence of increased Zn levels with increased Ca levels were confirmed. The pattern of metal contents encountered in some tumors was rather related to the biological state of the tissue than to morphogenesis or descent from any specific embryonic layer. Regressive transformation like calcification or liquefaction had a distinct bearing on the distribution of metals. PMID- 8237170 TI - Aluminum concentrations in serum of patients with intracranial tumors. AB - The concentration of serum aluminum was measured by electrothermal atomic absorption spectrometry in 71 patients with intracranial tumors. The data were compared with those from a reference group of 15 healthy adult individuals. The serum concentrations of aluminum were significantly increased (p < 0.01) in patients with malignant gliomas and meningiomas. The concentrations of serum calcium, known as an antagonist of aluminum, were normal in the same patients. PMID- 8237171 TI - Pseudo-cystic dilatation of suprapineal recess. Case report. AB - We report an odd case of marked enlargement of suprapineal recess in a patient with triventricular hydrocephalus. Radiological diagnostic steps and etiological theories are discussed. PMID- 8237172 TI - [Primary cerebral lymphoma. Current status of diagnosis and therapy]. AB - Primary cerebral lymphomas are rare tumors. In the last years the frequency of these space occupying lesions is increasing. Diagnosis by image-generating methods is often difficult. Stereotactic biopsy is a reliable diagnostic tool. Explorative craniotomy with tumor resection should be avoided if possible. An effective multimodal therapeutic regimen is the combination of radiotherapy and chemotherapy. Epidemiologic data, diagnostic methods and therapeutic proceedings are presented and discussed on a series of 7 patients with primary cerebral lymphomas treated in our department. PMID- 8237173 TI - [The postnatal development of porcine purine catabolism and the effect of the environmental temperature on turnover]. AB - Experiments in piglets indicate that the catabolic metabolism of purines is only developed during the first 4 weeks of life. During this time the catalytic activity of uricase in liver homogenates is increased about 5.5 fold (p < 0.01). The counts of uricase-carrying peroxisomes and cores are increased to a similar degree (p < 0.01). In the same time the catalytic activity of cytosolic xanthine oxidase is elevated about 4.6 fold (p < 0.01), mainly in the second and third week. In blood plasma uric acid is decreased from 19 to 12 molar % during weeks 1 to 4, hypoxanthine, xanthine and allantoin remain relatively constant. In urine xanthine and uric acid are decreased, hypoxanthine and allantoin are increased. At the age of 2-3 days a decrease of ambient temperature from 32 to 20 degrees C caused an increase of the plasma pool (p < 0.01) and turnover (p < 0.05) of uric acid. Pool size and turnover of hypoxanthine are only slightly elevated. The results are discussed in view of the slow development of hepatic enzymes, the increase of uric acid pool at lowered ambient temperature and causal events leading to urate nephropathia. PMID- 8237174 TI - Patterns of release of oxytocin, prolactin, insulin and LH in lactating sows, studied using continuous blood collection technique. AB - To describe the patterns of release of oxytocin, prolactin, insulin and LH in relation to suckling and to investigate relationships between them, blood was collected continuously from four sows at a rate of 1 ml per min for 12 h (0900 2100 h) on day 10 (D10) and day 20 (D20) of lactation. The 12-h period was divided into four smaller periods: P1 (0900-1200 h), P2 (1200-1500 h), P3 (1500 1800 h) and P4 (1800-2100 h). The number of nursings occurring during a 12-h period tended to decrease between D10 (17.3 +/- 0.3) and D20 (16.0 +/- 0.3). About 40-50% of all sucklings induced a significant rise in plasma oxytocin, while prolactin gradually increased following each suckling. Decrease in the levels of oxytocin (14.1 +/- 0.4 vs 7.1 +/- 0.4 pmol/l, P < 0.001), prolactin (12.9 +/- 0.3 vs 7.9 +/- 0.1 micrograms/l, P < 0.001) and insulin (24.8 +/- 1.7 vs 14.4 +/- 1.7 mU/l, P < 0.001) were observed between D10 and D20. Average and basal levels of LH and the number of LH pulses increased (P < 0.01) between D10 and D20. A negative correlation between number of sucklings and number of LH pulses was demonstrated on D20 (r = -0.59, P = 0.07). There was a positive correlation between levels of LH and oxytocin on D10 (r = 0.54, P = 0.001) and between prolactin and insulin levels on D20 (r = 0.34, P = 0.05). PMID- 8237175 TI - Ameloblastoma (adamantinoma) in a buffalo. AB - A six year old buffalo was admitted with a chronic indurated swelling at the left ramus of the mandible since 3 months. The radiographic picture showed a large osteolytic area and soft tissue swelling at the cheek teeth of the mandible. His topopathological examination disclosed neoplastic epithelial cells arranged in islands and cords. The cells were arranged in palisade form resembling the ameloblasts. The clinical findings, the radiographic features and the criteria of the neoplastic cells confirmed an ameloblastoma in a buffalo. PMID- 8237176 TI - Fate of the antinutritive proteins of soyabean in the ovine gut. AB - Defatted raw soyabean flour was administered intraruminally at a level of 10 g/kg body weight to a sheep fitted with ruminal, duodenal, and ileal cannulas. Soyabean lectin and trypsin inhibitor appeared in the duodenum within one hour, both reached the terminal ileum and disappeared within 24 hours. The passage of the lectin along the gut progressively lagged behind that of the inhibitor, which indicates that the lectin binds to the intestinal surface. The effect of the inhibitors was neutralized by a marked pancreatic hypersecretion. Contrary to earlier assumptions, the antinutritional proteins were not effectively degraded in the forestomachs. PMID- 8237177 TI - Perianal squamous cell carcinoma in goats. AB - Two cases of squamous cell carcinoma arising from the perianal skin of aged female Sannen crossbred goats are described. The occurrence of a tumour in this specific location is very rare, and the presence of the identical tumour in two animals of the same herd is unusual. PMID- 8237178 TI - Supplements of cod liver oil to lactating sows. Influence on milk fatty acid composition and growth performance of piglets. AB - Twelve sows received a supplement of 50 ml cod liver oil, without vitamin A and D, daily from day 107 of gestation until weaning. Eight sows served as control. A milk sample was obtained from each sow at parturition and on day 14 of lactation. The piglets were weighed at birth, on days 7, 14 and on the day of weaning. Eicosapentaenoic acid (20:5, n-3) (EPA), docosahexaenoic acid (22:6, n-3) (DHA) and docosapentaenoic acid; (22:5, n-3) (DPA) were found in milk fat from all sows. At parturition the contents of EPA, DPA and DHA were significantly higher in the group given cod liver oil supplement. After 2 weeks of lactation only EPA and DHA were elevated compared with the non-supplemented group. No effect on weight gain or on the overall morbidity of the piglets was found. The animal health in this farm was very good, and the growth rate faster than the average for Norwegian pigs. PMID- 8237179 TI - Physical performance of veal calves during chronic iron deficiency anaemia and after acute iron overload. AB - Treadmill experiments were performed with calves fed milkreplacer (MR) containing 8, 18, 52, or 86 mg of iron (Fe)/kg during the fattening period. Calves walked at 1 m/s for 10 min and then recovered for 15 min. Calves fed 8 and 18 mg of Fe/kg of MR developed severe hypoferraemia and anaemia whereas plasma (PFe) and haemoglobin (Hb) concentration, erythrocyte number (EC) and packed cell volume (PCV) in calves fed 52 and 86 mg of Fe/kg of MR were normal. PFe did not change, whereas Hb, EC, PCV, and blood lactate, heart rate (HR), respiratory rate (RR), respiratory minute volume (VE), fractional oxygen extraction rate (FO2) and O2 consumption (VO2) rapidly increased during walking, and then decreased during recovery. Hb, EC, PCV, partial O2 pressure of blood, FO2, and VO2 were lower, whereas HR, RR, respiratory exchange rate (RER) and blood lactate were higher in calves fed lowest as compared with those fed highest amounts of Fe, while these traits usually were intermediate in other groups. Calves fed 8 mg of Fe/kg of MR were in addition iv injected with FeOH for 3 d before walking. PFe, but not Hb, EC, and PCV, was drastically rised by FeOH. During walking, merely VO2 was improved, but only slightly. In conclusion, adjustments in calves with severe Fe deficiency anaemia were markedly impaired, and barely improved by acute elevation of PFe. PMID- 8237180 TI - Treadmill exercise of calves with different iron supply, husbandry, and work load. AB - Experiments were designed to study physiological responses of male veal calves to treadmill exercise, dependent on iron intake (20, 35, and 50 mg of iron/kg of milk powder, respectively). Calves were fattened from 75 to 180 kg BW and were either restrained or could freely move on straw litter. The ADG and feed utilization as well as haemoglobin and the plasma iron concentration were positively influenced by iron intake (P < or = 0.05). At the end of the fattening period, calves walked on a treadmill at 0.8, 1.1, or 1.3 m/s for 15 min. Resting values of all cardiorespiratory parameters were similar in all groups. During exercise, heart rate, respiratory rate, respiratory minute volume, and oxygen consumption increased and after 3 min reached steady-state values, which depended on work load. The except was respiratory rate (which reached a maximum already at the lowest speed) and oxygen extraction rate (which remained at basal level during exercise, but in part decreased after walking). In calves fed the least amount of iron, hence with the lowest haemoglobin concentrations, oxygen consumption was lower than in other groups. In addition, cortisol concentrations increased most markedly during exercise in calves fed the least amount of iron. PMID- 8237181 TI - Effects of concurrent oral administration of toxic levels of monensin and lead on concentration of different elements in the liver of broiler chicks. AB - Monensin and lead, separately or concurrently, were orally administered to broiler chicks at different toxic levels. Monensin slightly increased the selenium and profoundly increased the lead and iron levels of liver. Lead also increased the level of iron in liver. Levels of lead and iron in liver tissue further increased when monensin and lead were administered concurrently. An increased mortality was recorded due to concurrent administration of monensin and lead during acute toxicosis but during subacute toxicosis body weights were higher in birds administered monensin and lead concurrently than those given these substances separately. PMID- 8237182 TI - Mitogenic effect of lymph, amniotic and allantoic fluid in bovine fetal myoblasts. AB - Amniotic and allantoic fluids, as well as afferent lymph (tissue fluid), stimulated DNA synthesis in bovine fetal myoblasts (BFM) at low concentrations (5%). Higher doses abolished the mitostimulatory effect of these fluids and led toward inhibition of [3H]mTdR incorporation into DNA. These effects were much more evident (particularly in the case of amniotic fluid) when fluids were lyophylized prior to the experiment. BFM protein synthesis was accelerated by lymph in a dose dependent manner. PMID- 8237183 TI - Effect of a booster vaccination against influenza and equine herpes virus on cardio-respiratory adjustments to strenuous exercise and training in thoroughbred horses. AB - This study was conducted in order to assess whether exercise- and training induced cardio-respiratory adjustments are modified during the 10-day period which follows a booster vaccination with an oily adjuvanted inactivated vaccine against influenza and equine herpesvirus-1 (Equiffa). Nine healthy vaccinated thoroughbred horses were used. Six were revaccinated and three were kept as control. All the horses completed a standardised exercise test (SET) that was repeated 4 times, i.e. 10 (SET1) and 2 (SET2) days before revaccination, and 2 (SET3) and 10 (SET4) days after revaccination. During the whole experimental period the horses were trained 6 days per week according to an interval training schedule. Respiratory airflow, tidal volume, respiratory rate and expired minute volume (VE) were measured using a face mask and 2 ultra-sonic pneumotachographs. The oxygen uptake (VO2) and carbon dioxide production (VCO2) were obtained on a breath-by-breath basis, using a mass spectrometer. Heart rate (HR) was continuously measured with a polar horse tester. Venous blood was sampled before and after the test and analysed for biochemical determinations. Blood was also sampled before and 21 days after the booster vaccination for circulating antibody titration. The peak VO2, VCO2, ventilatory equivalent (VE/VO2) and oxygen pulse (VO2/HR) were significantly improved by the 3-week training period. The other cardio-respiratory parameters as well as most of the blood parameters remained unchanged throughout the 4 SETs. The revaccination did not impair any of the parameters measured before, during, or after the SETs. All revaccinated horses showed a rise in antibodies against influenza virus type A1 and A2 and EVH-1 compared to the control horses. It was concluded that, in our horses, revaccination with an oily adjuvanted inactivated vaccine against influenza and EVH-1 did not impair their cardiorespiratory and metabolic adjustments to strenuous exercise and intense training, and that the seroconversion due to revaccination was unaffected by the intense daily exercise experienced by the animals in the post-vaccination period. PMID- 8237184 TI - Desmitis of the accessory ligament of the deep digital flexor tendon in the horse: clinical and ultrasonographic features. A report of 24 cases. AB - The clinical and ultrasonographic features of desmitis of the accessory ligament of the deep digital flexor tendon (carpal check ligament) are reported in a series of 24 horses. Lameness associated with the carpal check ligament especially was found in Warmblood riding horses ranging in age from 6 to 18 years with a mean age of 11.3 years. The most striking clinical feature was a firm, painful swelling medially and laterally in the proximal and/or mid metacarpal region, corresponding with the dorsoproximal-palmarodistal oblique direction of the carpal check ligament. Sonographically swelling of the carpal check ligament was common, the enlarged ligament mostly showing a type 1-2 texture. Occasionally a more localized lesion with a type 3-4 texture was present. About 40% of the horses also showed additional lesions in (one of three other flexor tendons or ligaments in the metacarpal region. Most horses were treated conservatively, i.e. boxstallrest and/or a controlled exercise regime were applied. From the 19 horses available for follow up (1-36 months), only 3 horses resumed full work. Obviously the prognosis of this injury is guarded. PMID- 8237185 TI - Pentagastrin cytoprotection in ethanol-induced gastric mucosal lesions in rats. AB - To establish pentagastrin cytoprotection, the effectiveness of various doses of pentagastrin on ethanol induced gastric mucosal lesions was investigated in Wistar rats. Significant protection was obtained only after parenteral pretreatment with the exception of the lowest dose (1 microgram/kg b.w.). Pentagastrin cytoprotection is not mediated either by a dopamine, muscarinic or gastrin/CCK receptor or by prostaglandin synthesis. However, the protective effect of pentagastrin is abolished by prior vagotomy, although this procedure alone or sham operation is ineffective to influencing control-ethanol lesions. In secretory studies pentagastrin increased both the volume of gastric juice and total acid output. Unlike cytoprotection, these were reversed by vagotomy, but also with atropine and problumide, whereas domperidone and indomethacin were ineffective. PMID- 8237186 TI - Recording of intratracheal pressure in the horse under field conditions as a method for evaluation of upper airway resistance. AB - Recording of intratracheal pressure during exercise as a method for evaluating upper airway obstruction in the horse has previously been carried out on the treadmill. In the present study the method was applied to normal horses, outdoors under field conditions, to assess its usefulness for analysis of respiratory problems mainly occurring under racing conditions. The horses examined performed a standardized exercise test during which the intratracheal pressure was recorded with a tape recorder receiving amplified signals from a transducer connected to a percutaneously inserted intratracheal catheter. The exercise test was carried out under three different conditions: with (1) sulky or (2) rider outdoors and (3) on treadmill indoors. The latter test was included for comparison between field conditions and treadmill exercise. From the recordings obtained, several respiratory variables were calculated. The pressure graphs essentially resembled those obtained previously in tests performed exclusively on the treadmill. There were, however, some differences between the reactions of the horses to the three tests, which seem to necessitate a control material studied under comparable conditions. The technique is simple and the equipment for the outdoor tests is comparatively inexpensive, qualities which seem to make these tests suitable for studies of upper airway obstructions in clinical routine. Further, the outdoor tests may permit investigations of disorders of the respiratory tract that occur mainly under racing conditions. PMID- 8237187 TI - An outbreak of enterotoxaemia in suckling camels. AB - An outbreak of enterotoxaemia was observed for the first time in suckling camels in Saudi Arabia. The animals were weak, diarrhoeic and succumbed quickly to exertion. The main pathological findings were those of acute catarrhal enteritis and acute myocardial degeneration. Clostridium perfringens was isolated from the enteric lesions; Aeromonas hydrophila was also identified. The properties of both isolates were studied. PMID- 8237188 TI - Effect of serotonin on gastrointestinal electrical activity in the conscious piglet. AB - In conscious piglets with electrodes implanted in the wall of the antrum pylori, duodenum, jejunum and ileum, the influence of intravenous infusion of serotonin (5-HT), 4 micrograms/kg/min for 2 hours, on gastrointestinal electrical activity and arterial pressure was examined. 5-HT was without significant influence on antral electrical activity and arterial pressure. In the small intestinal parts 5 HT provoked a shortening of the interval of the migrating myoelectrical complex (MMC) by reducing the duration of phase I and phase II activities, and an increase in propagation velocity. 5-HT also induced an increase in phase III activity. A possible involvement of 5-HT in the initiation and propagation of the MMC is suggested. PMID- 8237189 TI - Evaluation of an enzyme linked immunosorbent assay for the determination of free thyroxine in canine plasma samples assisted by data on biological variation. AB - The purpose of the present study was to evaluate a commercially available enzyme linked immunosorbent assay (ELISA) for the determination of the free thyroxine (FT4) concentration in canine plasma samples. The test kit Enzymun-Test (Boehringer Mannheim) for determination of FT4 concentrations in human serum and plasma was used in connection with the analyzer Enzymun-Test(Boehringer Mannheim) System ES-33 by following the assay protocol suggested by the manufacturer. To assist in the evaluation of the ELISA method, goals for the precision of the assay were established from the inter- and intraindividual coefficients of variation estimated from determination of the plasma FT4 concentration in blood samples collected weekly from 8 healthy dogs for a period of 4 weeks. The intra- and interassay coefficients of variation ranged from 6% to 13%, the proposed goals for the precision ranging between 10% and 15%. The detection limit of the assay was 2 pmol/l which is well below the FT4 concentration commonly observed in plasma from dogs, and the FT4 concentrations determined in plasma from healthy dogs were comparable to those reported in the literature. Hence, the ELISA method evaluated in this study offers a precise and accurate measurement of the FT4 concentration in canine plasma samples, the inter- and intraassay precision comparing favorably to the proposed specifications for assay precision using estimates of the inter- and intraindividual coefficients of variation. PMID- 8237190 TI - The influence of storage time on the nucleotide concentration of canine platelets. AB - Nucleotide concentrations of canine platelets during storage for 50 days were determined to find out the variation in the concentration of various nucleotides in canine platelets during storage at +4 degrees C for 50 days. Platelets of 3 dogs were stored in whole blood in PVC bags containing CPDA-1 stabilizer. The bags were kept at 4 degrees C for 50 days. Changes in the energy metabolism of the platelets during storage were determined by successive measurements of the following nucleotides: adenosine-diphosphate (ADP), adenosine-tri-phosphate (ATP) and guanosine-tri-phosphate (GTP). In preparation for the measurement of their nucleotide content, the platelets were isolated from the blood at regular intervals and their membranes destroyed by deep freezing, thawing, double sonification in ice and protein precipitation with perchloric acid. The concentration of nucleotides within the platelets was determined by "High Pressure Liquid Chromatography" (HPLC). ADP, ATP and GTP were identified in the peak areas of the HPLC using 3 different techniques: comparison of retention time of standard nucleotide solutions as control samples, enzyme tests and thin layer chromatography. Adenosine-mono-phosphate (AMP), adenosine, hypoxanthine and inosine mono 5'-phosphate (IMP) were not found in dog platelets. Platelet ATP concentration decreased significantly during 50 days of storage (p < 0.001), and was reduced to 24% of the concentration measured after 1 hour of storage on day 50. ADP concentration also dropped significantly during 50 days of storage (p < 0.001). On day 50 the average ADP concentration was reduced to about 34% of the concentration measured after 1 hour of storage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237191 TI - Influence of naproxen on the zinc, copper, magnesium and calcium content of guinea pig tissues. AB - The effect of the administration of chronic doses of naproxen on the zinc, copper, magnesium and calcium concentrations in guinea pig tissues was studied. One hundred mg naproxen/kg body weight/week were administered intraperitoneally to adult female guinea pigs in two doses twice a week for eight weeks. A statistically significant decrease was found of the zinc content in the brain, lung, heart, liver, kidney, spleen and adrenals, of the copper content in the liver, kidney and adrenal and of the magnesium content in the spleen and heart. A significant increase was observed of the calcium content in the adrenals, heart, spleen and uterus of the treated animals, while no significant changes were observed in the thigh bone concentrations of the various ions tested. PMID- 8237192 TI - Dynamics of porcine humoral responses to experimental infections by Spanish Trichinella isolates: comparison of three larval antigens in ELISA. AB - Five groups of six helminth-free pigs of the Iberian and Landrace x White breeds were each experimentally-infected randomly at a dose of 150 larvae per kg, body weight with one of the five Trichinella isolates, coded as Gm-1, Co-77, Laso, C 76 and Mad-83. These isolates were selected from the two Trichinella species (T. spiralis and T. britovi) so far identified in the Iberian Peninsula. Specific antibodies against the infections were measured on days -14, 0, 6, 16, 20, 27, 34, 49, 63 and 82 post-infection (p.i.), in a comparative indirect ELISA assay using three different antigens prepared from the muscle stage of the parasite. The antigens were a crude saline larval extract (CSLE), excretory-secretory (ES) and surface stripped cetyl-trimethyl ammonium bromide detergent (CTAB) products. Over all, the dynamics of antibody responses were very similar for infections with the five different isolates, although a significant delay in positive and maximum antibody titres was seen in the group infected with C-76, the isolate exhibiting a marked low infectivity to domestic mammals. ELISA with the crude antigen was more sensitive (positive antibody titres appeared between days 6 and 16 p.i.) than with the ES and CTAB preparations (positive titres appeared between days 16 and 20 p.i.). For infections with the C-76 isolate, positive titres appeared between days 16 and 20 p.i. with the CSLE antigen and, on day 27 p.i. with the ES and CTAB antigens. Little cross-reactivity with other common porcine helminth infections appeared only when ELISA was carried out with CSLE and CTAB antigens. PMID- 8237193 TI - Comparison of different serological methods for the determination of antibodies to Chlamydia psittaci in pigeon sera. AB - Several methods for detecting antibodies to Chlamydia psittaci using sera from pigeons were compared with regard to their sensitivity, specificity and efficacy. The reference method used was indirect immunofluorescence (IFI), which uses as antigen intracellular inclusions in McCoy cell monolayers. Indirect micro immunofluorescence (MIF) using as antigen C. psittaci cultured on yolk sacs of embryonated eggs, direct and indirect complement fixation tests (CFT and ICFT, respectively), and enzyme-linked immunosorbent assay (ELISA) were compared to the reference method. Indirect micro-immunofluorescence proved to be the most efficient method, while ELISA was the most sensitive, though showing a very low specificity. No statistically significant difference was found in comparing the two complement fixation tests. The use of ICFT to check for the negative results obtained in CFT is questioned. Possible reasons for the different results with the methods used are discussed. PMID- 8237194 TI - NaCl-tolerance of Campylobacter isolates from birds and Campylobacter type strains and variation of their serological behaviour. AB - Growth on media containing 1.5% NaCl is one of the criteria for phenotypical differentiation of Campylobacter laridis from other thermophilic Campylobacter spp. Campylobacter isolates from birds and Campylobacter type strains could be adapted to growth at 3% NaCl within 19 to 72 subsequent passages on nutrient agar with increasing salt contents. The acquisition of salt-tolerance was stable after ten passages on media without salt and did not induce changes in other phenotypical characteristics. The results of slide agglutination demonstrate changes in the antigenic pattern of the Campylobacter strains after growth in salt. Heat-labile and heat-stable antigens of the salt-tolerant variants of Campylobacter type strains differed from those of the parent strains. PMID- 8237195 TI - [Comparison of two polysaccharide antigens for the serological diagnosis by ELISA of porcine pleuropneumonia (serotype 5)]. AB - Two antigens of polysaccharidic nature were evaluated for their specificity in serodiagnosis of porcine pleuropneumonia (serotype 5). Recently, long chains lipopolysaccharides (LC-LPS) was shown to increase the specificity of the ELISA test for A. pleuropneumoniae serotype 5 compared to a saline extract of boiled formalinized whole cells. Nevertheless, some cross reactions were noticed with LC LPS. We have thus cleaved the LPS molecule at the ketosidic bond between the lipid A and the core. The lipid A was separated from the polysaccharide (PS) by centrifugation. With an indirect sandwich ELISA, it was possible to demonstrate that the PS yielded positive responses with homologous sera (serotype 5) and negative responses with heterologous sera (serotype 3 and undeterminated serotype). Overall, the use of the PS did not notably increase the specificity of the ELISA test. Since production of PS from LC-LPS and its separation from the lipid A is not simple, we do not recommend its use in serodiagnosis. PMID- 8237196 TI - Uterine and placental alterations in pregnant sows associated with the porcine epidemic abortion and respiratory syndrome (PEARS). AB - In the winter of 1990/91 a new, economically devastating disease occurred in european pig breeding herds, characterized by late-term abortion, stillbirth and a high morbidity and mortality of suckling piglets. Because of the clinical picture the disease was named porcine epidemic and respiratory syndrome (PEARS). In this study investigations were carried out in tissues of uterus and placentae of late gestational sows (107 to 112 days of gestation) in three different groups of animals: group I = control animals (n = 2) group II = naturally infected sows (n = 12) from farms, where PEARS recently had been introduced; group III = sows (n = 2), experimentally inoculated with placental homogenates from animals of group II. Both in naturally infected as well as in experimentally infected sows a multifocal, lymphohistiocytic vasculitis and perivascular cell infiltration was observed in the endometrium and maternal part of the placenta, but not in the fetal one. In the fetomaternal unit there were multifocal microseparations of the epithelial layers present. Transmission electron microscopically spherical or oval virus-like particles of 45 to 75 mm in diameter were frequently found on the surface of endothelial cells of blood vessels in the maternal placenta, in a few cases in the intercellular channel system between uterine epithelial cells or on endothelial cells of capillaries in the fetal placenta. Serological results indicate, that an infection with Lelystad virus had occurred in naturally and experimentally infected sows and that transplacental infection was present. PMID- 8237197 TI - A close comparative study on the response of sheep and goats to experimental orf infection. AB - A comparative experimental study of the clinico-pathological and serological response of sheep and goats to orf infection was made. All infected animals depicted typical orf lesions and seroconverted. The clinical, serological, gross and histopathological responses were similar in both species. However, the disease span was longer in sheep than in goats. It was concluded that sheep could play an important role in maintenance of the virus in the environment. PMID- 8237198 TI - Experimental aspergillosis in goats: clinical, haematological and mycological studies. AB - Thirty-one (including ten lactating) healthy goats were divided into 3 groups viz. intravenous (I/V), intratracheal (I/T) and intramammary (I/M) on the basis of route of inoculation of Aspergillus fumigatus spores. Two animals were kept as control for each group. Clinical aspergillosis developed and was more severe and fatal in consequence in I/V group than those of other groups. Respiratory symptoms appeared and persisted in both I/V and I/T groups. Acute mastitis developed in the right infected udder halves only with a brief and transitory illness in all goats of I/M group and marked discrepancy in size between right inoculated and left uninfected udder halves was observed. Haematological examination revealed significant decrease of haemoglobin concentration and significant increase in both TLC and DLC (neutrophilia) in I/V group but no marked haematological change was observed in I/T and I/M groups. Mycological isolation in pure culture, demonstration of fungus in the impression smears by lactophenol cotton blue staining and in histological sections by PAS and GSM methods were achieved in the lungs, kidneys, heart, liver, spleen and abomasum in I/V group, in the lungs and occasionally in the mediastinal lymph node and liver in I/T group and in the right infected udder halves and their milk/secretions in I/M group. PMID- 8237199 TI - Prevention of cutaneous myiasis caused by screw worm larvae (Cochliomyia hominivorax) using ivermectin. AB - During March of 1991 and 1992, four trials were carried out in the central area of Argentina to evaluate the efficacy of ivermectin in the prevention of myiasis caused by Cochliomya hominivorax larvae. In the first trial 24 steers were artificially wounded, in the second and third trial, 36 and 20 bull calves were castrated while in the fourth trial, 30 newborn calves were used. On day 0, half of the animals of each trial was treated with ivermectin (200 micrograms/kg) and the rest remained as untreated controls. None of the treated animals sustained a screw-worm larva infestation, but 3 of 12 (25%), 8 of 18 (44%), 5 of 10 (50%) and 8 of 15 (53%) of the controls developed active myiasis in trials 1 to 4 respectively. PMID- 8237200 TI - In borna disease virus infected rabbit neurons 100 nm particle structures accumulate at areas of Joest-Degen inclusion bodies. AB - Borna disease virus infected rabbits were chosen to search for electronmicroscopic structures. Intensively investigated hippocampal neurons showed intranuclear inclusions; 100 nm particle-like structures surrounded by 20 nm granular forms were prominent. In connection with elsewhere reported in situ hybridization studies of virus-specific RNA to areas of the Joest-Degen inclusions we suggest that these particle structures may represent Borna virus. Jost-Degen (8) found intranuclear inclusion bodies in neurons to be pathognomonic for Borna disease (BD) in the horse. Half a century later these structures were suggested to represent BD virus (BDV)-specific antigen aggregates (15). A century later we characterized the virus to contain a single and negative stranded RNA of 8.5 kb, which transcribes in the nucleus (5) and could show that virus complementary RNA seems to hybridize spot-like to nuclear areas, probably representing the Jost-Degen inclusions (7). Electron microscopic (EM) findings on structures in BDV infected brain cells and about particle-like structures obtained from infected tissue culture cells have been reported by different groups (1, 2, 3, 4, 5, 6, 9, 11, 12, 13). The demonstration of crystalline aggregates and filament bundles in the cytoplasma and karyospheridia (nuclear bodies) were prominent. Such structures were seen in infected rabbits, hamsters, rats mice and naturally infected horses. The phenotypic description of filamentous structures, crystalline aggregates in the cytoplasma and large karyospheridia were prominent. Based on our experience with BDV infections in rabbits we selected this species for ultrastructural studies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237201 TI - Natural Borna disease in domestic animals others than horses and sheep. AB - In 24 cats (Uppsala, Sweden) with neurological signs of "staggering disease" and typical neuropathology, 44% had Borna disease virus (BDV)-specific antibodies. In 173 cat sera (Berlin, Germany) of animals with unknown record, 7% were BDV positive. Out of 24 cats with undefined neurological disorders, 13% were BDV positive. Similarities in staggering disease of cats and Borna disease of horses and sheep suggest related etiological agents. PMID- 8237202 TI - [Serologic diagnosis of avian salmonelloses: adjustment of an ELISA test using antigens adsorbed with the aid of anti-colibacillary sera]. AB - A serological ELISA test for diagnosis of avian salmonellosis infections with Salmonella typhimurium or enteritidis has been established. Plates were coated half with a negative antigen and half with a positive antigen. Both negative and positive antigens were adsorbed with a monovalent agglutinant anti Escherichia coli serum prior to being distributed into wells of the plates. Sera from SPF birds and from SPF birds vaccinated and/or inoculated with numerous viruses or bacteria, or sera from conventional birds bacteriologically free of any salmonellosis were tested and the percentage of false positive reactions was inferior than 1%. In groups of birds naturally infected or experimentally inoculated with Salmonella enteritidis or typhimurium the percentages of positive individuals were ranged from 15 to more than 90% according to the doses and routes of inoculation and the delay between contamination and sampling. PMID- 8237203 TI - A case of bovine mastitis caused by Campylobacter jejuni. AB - A Campylobacter jejuni mastitis in a Holstein cow, 60 days into the first lactation, is reported. This was characterized by a sudden onset, a pyrexia, painful quarter and pink milk with a few small clots present. There was good response to a treatment protocol of: parenteral oxytetracycline, frequent stripping and intramammary infusions of erythromycin. This organism has a zoonotic property. PMID- 8237204 TI - Important endoparasites in sheep and goat flocks in Jordan. AB - In spring 1992 in Jordan 70 flocks of sheep and goats were investigated coproscopically. 838 of altogether 882 small ruminants (= 95.01%) harboured endoparasites. In three different parts of the country--in the vicinities of Ma'an, Kerak and Amman--the results were similar (Dictyocaulus, Nematodirus, Moniezia) or they differed (protostrongylids, Marshallagia and other trichostrongylids, Eimeria). In connection with the established parasite status remarks concerning the therapy, including the problems involved as well as remarks concerning the time of treatment in the investigated area will be made. Finally, suggestions will be made for important and specialized follow-up studies regarding improvement of the situation in this country. PMID- 8237205 TI - Papovavirus-induced trichogenous tumours in Syrian hamsters (Mesocricetus auratus). AB - Tumorous, virus-induced skin lesions in two golden hamsters (Mesocricetus auratus) were characterized macroscopically and by means of light- and electron microscopy. Evidence of a virus was demonstrated in the ultra-thin sections and by the negative staining method. The morphological findings confirm the assumption that infections with papoviruses--probably of the polyomavirus genus- were involved. PMID- 8237206 TI - Use of a feline panleukopenia modified live virus vaccine in cats in the primary stage of feline immunodeficiency virus infection. AB - Cats in the primary stage of Feline Immunodeficiency Virus (FIV) infection and FIV seronegative cats were vaccinated with a modified live Feline Panleukopenia virus vaccine (FPV-MLV). The FPV-MLV strain was not pathogenic for FIV seronegative cats, whereas in FIV infected cats it produced severe clinical signs, similar to those typically observed in cats naturally infected with field strains of FPV (fever, diarrhoea, leukopenia). PMID- 8237207 TI - Experimental encephalomyocarditis virus infection in pigs. AB - A field isolate of Encephalomyocarditis (EMC) virus was inoculated intravenously into 8 pigs. Four animals died at post inoculation day (PID) 2, the remaining being sacrificed at PID 5, 7, 11 and 15. Two control, in-contact pigs were sacrificed at PID 19. Virus was isolated from leucocytes and nasal swabs until PID 4, from rectal swabs until PID 2 and, in the pigs found dead at PID 2, from several organs. EMC virus was further isolated from brain and spleen of the pig sacrificed at PID 7. One of the 2 control pigs became infected: virus was isolated from nasal swabs at days 6 and 7 and from leucocytes at day 4 of the experiment. Serum-neutralizing (SN) antibody was detected in the injected pigs starting from PID 4; two days later, it was also revealed in the infected, in contact control. To our knowledge, this is the first report of an experimental transmission of EMC virus infection in pigs by contact exposure. PMID- 8237208 TI - Efficacy of various non-oily adjuvants in immunization against the Aujeszky's disease (pseudorabies) virus. AB - Standard oil and various non-oily adjuvants were compared for use in immunization against the Aujeszky's disease (pseudorabies) virus, both in mice and swine, and using either inactivated virions or purified glycoproteins as antigen. Mineral oil, sodium alginate, aluminium hydroxide, and saponin were assayed in mice as adjuvants for inactivated virions, saponin being the most efficient. The addition of Mab anti-CD3 did not improve either immune response or protection achieved in mice using viral particles with oil or sodium alginate. When purified glycoproteins were used as antigens, the use of ISCOM greatly enhanced specific T cell responses and protection of mice. The incorporation of Mab anti-CD3 into ISCOM conferred 100% protection of mice. Surprisingly, when an ISCOM containing glycoproteins was assayed in swine in a single-dose trial, no improvement on the protection conferred by the oily adjuvant was observed. PMID- 8237209 TI - A sporulation gene in Coxiella burnetii? AB - During a search for conserved target sequences on the genome of Coxiella burnetii for a diagnostic PCR a NotI/EcoRV DNA fragment was cloned and sequenced from the isolate "Nine Mile", Phase I (sequence data are registered under accession number X70045 in data bases EMBL, GENEBANK and DDBJ). On this fragment we have found a sequence of 1741 base pairs which showed an exceptionally high homology (60% on the nucleic acid level and 49.6% on the amino acid level) to the sporulation gene "spoIIIE" of the gram-positive bacterium Bacillus subtilis. This is first evidence of the molecular basis for formation of spores in Coxiella burnetii which has been postulated in the literature by electron microscopic investigations. PMID- 8237210 TI - Prolonged persistence of cytopathogenic bovine viral diarrhea virus (BVDV) in a persistently viremic cattle. AB - A bull persistently viremic with noncytopathogenic (ncp) BVDV was inoculated with the cytopathic (cp) BVDV strain TGAC, which had been found to be antigenically different from the endogenous ncpBVDV (ncpW8). Neutralizing antibodies against strains NADL and TGAC were detectable 12 days and four weeks post infection, respectively. The animal developed fever and diarrhea 15 weeks post infection. On days 3 and 8 after onset of diarrhea a cpBVDV (cpX) was isolated from feces. Antigenic analysis using monoclonal antibodies (MoAbs) showed that cpX and the endogenous ncpBVDV (ncpW8) had identical reactivity patterns except for one epitope that was neither expressed on TGAC nor on ncpW8. Using polymerase chain reaction analysis it was shown that both TGAC and cpX contained a p8 phi gene duplication combined with genomic insertions of identical size. Restriction enzyme analysis of the TGAC and cpX amplicons using four enzymes showed an identical cleavage pattern, except for HaeIII digestion where an additional fragment was observed with cpX. These results suggest that cpBVDV strain TGAC persisted in the viremic animal and apparently caused disease after 15 weeks. PMID- 8237211 TI - Comparison of the use of UV light and nitrosoguanidine as mutagenic treatments in Aspergillus parasiticus. AB - In the present work we study two different mutagenic methods in Aspergillus parasiticus: UV light and Nitrosoguanidine treatments. With UV light we obtained more suitable results, with survival percentage around 1% in many experiments. With Nitrosoguanidine our results were around 10% survival rate. We suggest the use of UV light to obtain mutants from A. parasiticus. PMID- 8237212 TI - Further evidence for the involvement of SmI cortical neurons in nociception: their responsiveness at 24 hr after carrageenin-induced hyperalgesic inflammation in the rat. AB - In this electrophysiological study, the responsiveness of neurons in the primary somatosensory cortex (SmI) was analyzed in rats with carrageenin-induced hyperalgesia for 24 hr. The functional implication of some changes in neuronal activity was improved in a few cases by a pharmacological test with Xylocaine injection in or close to the neuronal receptive field (RF), or with systemic aspirin. Unit recordings were performed alternately in the SmI cortex contralateral (Cc) or ipsilateral (Ci) to the inflamed hindpaw. In 29 rats with hyperalgesia tested prior to the recording session, 218 cells (128 in the Cc, 90 in the Ci) were tested with mechanical stimuli. In each SmI cortex, about 50% of them were driven by the stimulus. The "nonresponsive" neurons exhibited a higher firing rate in the Cc than in the Ci. The "responsive" (i.e., the somatosensory) neurons were classified according to their response to light touch, pinch, or joint movement. There was a highly significant difference between the two cortices, essentially because of the high proportion of "joint" Cc neurons (27 of 73 [37%] of the somatosensory neurons in the Cc, vs. only 8 of 47 [17%] in the Ci). "Light touch" neurons (41 of 73 [56%] in the Cc, vs. 35 of 47 [74.5%] in the Ci) had small RFs contralateral to the recording site. Of the 41 Cc cells of this type, 23 did not exhibit the classical characteristics of "light touch" cells; in particular, they exhibited striking discharges triggered by the stimulus but outlasting the stimulus duration, or occurring without intentional stimulation. These abnormal discharges were depressed or suppressed by injection of a local anesthetic (Xylocaine) in or close to the neuronal RF. "Pinch" neurons were very rare (5 of 73 [7%] in the Cc, vs. 4 of 47 [8.5%] in the Ci). Responses elicited from the inflamed paw were more pronounced than those from the noninflamed paw. "Joint" neurons were more numerous in the Cc than in the Ci. In addition, their responses obtained from contralateral RFs, and therefore from the inflamed paw, were more sustained than Ci responses elicited from the noninflamed paw. Afterdischarges of Ce neuronal responses and spontaneous paroxysmal activity were common on this side and were depressed by local anesthetic (Xylocaine) in their RFs or by systemic aspirin. These electrophysiological data emphasize the implication of SmI cortex in inflammatory hyperalgesia and more generally in pain processing.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8237213 TI - Effects of attention on visuomotor activity in the premotor and prefrontal cortex of a primate. AB - We examined neuronal activity in the primate premotor (PM) and prefrontal (PF) areas during a demanding spatial matching task. On each behavioral trial, a rhesus monkey moved its forelimb when a visual stimulus, called the "prime stimulus," reappeared at a previously cued location. Because it triggered a movement, the part of space cued by the prime stimulus had to be either remembered or attended during the time between prime stimulus presentations. Between the first and second appearances of the prime stimulus, behaviorally irrelevant visual stimuli could appear at one or several locations other than that of the prime stimulus. We could thereby examine the activity that followed a stimulus when it was attended versus when it was irrelevant and presumably unattended. We found that visuospatial attention affected neuronal activity in both the motor and "nonmotor" parts of the frontal cortex. The magnitude of attention effects exceeded that previously reported--a finding that probably resulted from the intensive attentional demands of the present task. PMID- 8237214 TI - Barrels V: proceedings of a satellite symposium of the 1992 Society for Neuroscience meeting. PMID- 8237215 TI - Comparison of the connectional properties of the two forelimb areas of the rat sensorimotor cortex: support for the presence of a premotor or supplementary motor cortical area. AB - The existence of multiple motor cortical areas that differ in some of their properties is well known in primates, but is less clear in the rat. The present study addressed this question from the point of view of connectional properties by comparing the afferent and efferent projections of the caudal forelimb area (CFA), considered to be the equivalent of the forelimb area of the primary motor cortex (MI), and a second forelimb motor representation, the rostral forelimb area (RFA). As a result of various tracing experiments (including double labeling), it was observed that CFA and RFA had reciprocal corticocortical connections characterized by preferential, asymmetrical, laminar distribution, indicating that RFA may occupy a different hierarchical level than CFA, according to criteria previously discussed in the visual cortex of primates. Furthermore, it was found that RFA, but not CFA, exhibited dense reciprocal connections with the insular cortex. With respect to their efferent projection to the basal ganglia, it was observed that CFA projected very densely to the lateral portion of the ipsilateral caudate putamen, whereas the contralateral projection was sparse and more restricted. The ipsilateral projection originating from RFA was slightly less dense than that from CFA, but it covered a larger portion of the caudate putamen (in the medial direction); the contralateral projection from RFA to the caudate putamen was of the same density and extent as the ipsilateral projection. The reciprocal thalamocortical and corticothalamic connections of RFA and CFA differed from each other in the sense that CFA was mainly interconnected with the ventrolateral thalamic nucleus, while RFA was mainly connected with the ventromedial thalamic nucleus. Altogether, these connectional differences, compared with the pattern of organization of the motor cortical areas in primates, suggest that RFA in the rat may well be an equivalent of the premotor or supplementary motor area. In contrast to the corticocortical, corticostriatal, and thalamocortical connections, RFA and CFA showed similar efferent projections to the subthalamic nucleus, substantia nigra, red nucleus, tectum, pontine nuclei, inferior olive, and spinal cord. PMID- 8237216 TI - Tactile-evoked response of sensory fibers in buccal and submandibular regions of the rat. AB - Evoked neural responses to tactile stimulation were recorded electrophysiologically from the mechanoreceptive afferent fibers innervating the buccal and submandibular regions of Wistar rats anesthetized with sodium thiopental. Miniature probes 200 microns in diameter were used, and data analysis was performed on the mechanosensitivity of responses to tactile stimulation in the areas innervated by the mental, mylohyoid, auriculotemporal, and cervical nerves. Mechano-sensitivity of each area showed a characteristic distribution of slowly adapting (SA), rapidly adapting (RA), C-fiber (CF), and hair follicle (HF) units in individual receptive fields. The density of the SA units was high in the areas innervated by the mylohyoid and auriculotemporal nerves. The CF units were concentrated in the small dome in the area of the mylohyoid nerve and the auriculotemporal nerve, as shown by a significant response to the dynamic features of stimulation. Estimation of the current needed for tactile acuity suggests an important role of the SA fibers in the areas innervated by the auriculotemporal, mylohyoid, and cervical nerves. PMID- 8237217 TI - Tactile-vibration-activated foci in insular and parietal-opercular cortex studied with positron emission tomography: mapping the second somatosensory area in humans. AB - Positron emission tomographic measurements were used to study the distribution of focal changes in cerebral blood flow (CBF) induced by vibrotactile stimulation of the hands and feet in 22 normal humans. Subjects received bolus intravenous saline injections containing approximately 60 mCi 15O-labeled water. Active regions during stimulation were defined relative to resting, nonstimulated states. Scan data from different subjects were averaged after stereotactic standardization. The results identified previously described foci of increased CBF in postrolandic sensory cortex (primary somatosensory cortex) and supplementary motor cortex. New statistical testing procedures provided independent demonstrations of two additional increases in regional CBF, bilaterally, within the sylvian fissure. One site along the parietal operculum corresponded to previous conjectures about a second somatosensory cortical area (SII) in humans. Another site also was found on the insula. No topographic organization was found in either location. The discussion considers these responsive areas to innocuous tactile stimuli in reference to suggestions about a role for SII in the perception of pain. PMID- 8237218 TI - Physiological and morphological characteristics of spinal neurons projecting to the parabrachial region of the cat. AB - Neurons in the lumbosacral, superficial spinal dorsal horn in the cat were recorded extra- and intracellularly, using dorsal root stimulation as a search stimulus. Isolated neurons were tested for antidromic activation from the contra- and ipsilateral parabrachial region. Seventy-one nociceptive-specific neurons, 11 innocuous cooling neurons, and 8 multireceptive neurons were antidromically activated from the lateral parabrachial region. The receptive fields and response properties were typical of other lamina I and lamina II neurons, in that the receptive fields were usually discrete and relatively small, and the responses ranged from sluggish and decrementing to brisk and augmenting with afterdischarge. The conduction velocity to the parabrachial region averaged 3.7 m/sec for the nociceptive-specific neurons, 3.9 m/sec for the innocuous cooling neurons, and 13.5 m/sec for the multireceptive neurons. Intracellularly labeled neurons were mostly medium to large Waldeyer-like neurons in lamina I. Some had axon collaterals that distributed varicosities in laminae I, II, and V. These data indicate that a slowly conducting nociceptive-specific and thermoreceptive pathway exists between the superficial dorsal horn and the parabrachial region at the pontine-midbrain junction. PMID- 8237219 TI - Afferent modulation of warmth sensation and heat pain in the human hand. AB - The hands of 14 normal humans were used to determine the somatotopic organization of the modulation of warmth sensation and heat pain by different forms of cutaneous stimuli. Test stimuli were 5-sec heat pulses ranging from 36 degrees to 51 degrees C, delivered to the fingerpads of digits 1, 2, 4, and 5 with a contact thermode. Conditioning stimuli (15 sec) bracketed the test stimuli and included vibration, noxious and innocuous heat, cold, and electrical pulses delivered to the fingerpads of digits that were adjacent or nonadjacent to the tested digits. Noxious (48 degrees +/- 1.3 degrees C), but not innocuous (43 degrees C), heat stimuli increased the perceived magnitude estimation of innocuous test stimuli (36-43 degrees C) by 20-37% when delivered to adjacent, but not to nonadjacent, digits. No other conditioning stimuli had any effect on the intensity of warmth perception. In contrast, both noxious and innocuous heat or electrical conditioning reduced the magnitude estimation of noxious (50-51 degrees C), but not innocuous, test pulses by 12-22% when delivered to adjacent digits. Conditioning of nonadjacent digits was significantly less effective. The analgesic effects of noxious and innocuous conditioning were approximately equal. Vibratory (120 Hz, 3.5 microns) and cold (15 degrees C) conditioning stimuli were ineffective. The results are consistent with a dermatomal somatotopic organization of tactile and heat modulatory influences on warmth sensation and heat pain. The results further suggest that the neural mechanism subserving warmth mediate a negative feedback influence on heat pain intensity. PMID- 8237220 TI - Tectal and related target areas of spinal and dorsal column nuclear projections in hedgehog tenrecs. AB - The terminal distributions of spinal and dorsal column nuclear projections to tectum, pretectum, and central gray of hedgehog tenrecs (Echinops telfairi and Setifer setosus) were investigated using anterograde axonal flow and various tracer substances. In the inferior colliculus, the densest and most extensive mesencephalic projections were found within the pericentral regions. One target area, referred to as the external portion of the inferior colliculus, was represented as a semicircle of grain patches lateral and caudal to the central nucleus. This region received somesthetic afferents from the dorsal column nuclei and from spinal segments at various levels. In contrast, after high cervical injections, the pericentral portion dorsomedial to the rostral half of the central nucleus was labeled almost exclusively. This area of labeling was distinct from the labeling in the central gray and might be best compared with the intercollicular zone in other species. The superior colliculus received projections predominantly from the high cervical cord; minor projections also arose from lumbar spinal segments and the dorsal column nuclei. The terminal field covered roughly the caudal half of the colliculus and involved the stratum griseum intermediale in a patch-like fashion. Some labeling was also found in the stratum griseum profundum and in the stratum griseum superficiale. Other than in the colliculi, weak pretectal projections were observed following dorsal column nuclear injections, while the nucleus of Darkschewitsch was labeled best following lumbosacral injections. All mesencephalic target areas were labeled consistently on the contralateral side, while their ipsilateral side was involved to a varying degree: The relatively most prominent ipsilateral labeling was seen in the central gray, being roughly similar on both sides; scarcely any labeling was noted in the ipsilateral superior colliculus. Tectal injections of retrograde tracer, in addition, revealed a considerable number of labeled neurons in a relatively cell-poor region immediately ventral to the high cervical dorsal horn. This region might correspond to the lateral cervical nucleus, an aggregation of neurons that so far has only been demonstrated in higher mammals. PMID- 8237221 TI - [Studies on gap junction formation and junctional communication between cultured endothelial cells and smooth muscle cells]. AB - Gap junction (GJ) between two contacted cells is the structural basis of intercellular communication. Gap junction transfer plays an important role in regulation of cell proliferation. Cultured endothelial cells (EC) and smooth muscle cells (SMC) were injected with fluorescein Lucifer Yellow by iontophoretic injection, and examined under fluorescein microscope. We have demonstrated that there existed homologous and heterologous GJ and contact mediated dye transfer between cultured EC and SMC. LDL and insulin which promote SMC proliferation may inhibit the junctional communication between SMC and EC. Tumor promoter TPA induced almost complete inhibition of junctional communication between EC and SMC. The results indicate that normally regulative signals of metabolism and growth are transferred between EC and SMC via junctional communication to maintain the vessel wall homeostasis. High dosage LDL and insulin may keep SMC away from normal growth control and induce their proliferation by means of inhibiting junctional communication. It may be one of important mechanisms of atherogenesis. PMID- 8237222 TI - [The roles of monocytes and the interaction between monocyte and T cell in human immune suppression induced by trichosanthin]. AB - This paper deals with the roles of monocytes and the interaction between T cell and monocyte in Trichosanthin-induced human immune suppression. The data strongly argue that (a) monocytes could mediate the immune suppression of Trichosanthin, but T cells alone could not; (b) T cells acquire the ability to mediate the suppression effect if they were pulsed with Trichosanthin together with monocytes; (c) Antipain, a blocking agent against antigen processing and presentation, could diminish the suppressive activity of the Trichosanthin-pulsed monocytes. The results suggested the dependence of Trichosanthin on monocyte in immune down-regulation. One of the mechanisms might be that Trichosanthin induces an inhibitory network through antigen processing and presentation by monocytes. PMID- 8237223 TI - [Biological characterization of a monoclonal antibody TB 21 against human transforming growth factor-beta 1]. AB - A hybridoma TB 21 clone was derived from fusion between Sp 2/0-Ag 14 myeloma cells and spleen cells of BALB/c mice immunized with transforming growth factor beta 1 (TGF-beta 1) purified from human platelets. The TB 21 clone was identified to produce monoclonal antibody with IgG1 subclass and had sufficient titer for immunoreactivity to both human platelets-derived TGF-beta 1 and recombinant human TGF-beta 1 by enzyme-linked immunosorbent assay (ELISA). Western blotting studies demonstrated that two immunoreactive bands corresponding to 25 Kda and 12.5 Kda molecules were observed in the sample of acid/ethanol extracts from human platelets. The affinity constant (Kaff) was determined as 1.47 x 10(8) M-1 with non-competitive ELISA. Moreover, using bioassay for the effects of TGF-beta 1 on the growth of mink lung epithelial cells (CCL/64 cell line) and fibroblast cells (NRK 49 F cell line), TB 21 IgG was shown to be able to neutralize the action of TGF-beta 1 on the growth of these target cells. Therefore, this monoclonal antibody may be a useful probe for studying the growth modulatory activity of TGF beta 1 in a variety of cells and tissues. PMID- 8237224 TI - The Angelchik prosthesis in the surgical therapy of reflux oesophagitis. Short and long term results. AB - The authors report the results acquired from 1981 to 1985 using the Angelchik prosthesis in 22 patients with reflux oesophagitis resisting medical therapy for more than 6 months. Clinical evaluation has been done using Visick scale, as modified by Deakin. Clinical and instrumental controls have been done at 1 month, 12 months and 5-10 years after the operation. Intraoperative mortality and morbidity were zero. Complications directly related to the operation were: transitory dysphagia in 4 cases, persistent dysphagia in 3 cases, transitory gas bloat syndrome in 1 case and persistent gas bloat syndrome in another, persistent stenosis in 1 case and prosthesis dislocation in 6 cases. Clinical results were satisfactory (success rate: 90.9%) only in patients with a short term follow-up (30 days). The full rate of success has come down to 84.2% and 66.6% in patients having follow-ups respectively of 1 year and 5-10 years. The results are truly disappointing and the authors conclude that this procedure is not to be employed. PMID- 8237225 TI - Surgical treatment of duodenal diverticula. AB - Fourteen patients with duodenal diverticula are reviewed. The various aetiopathogenic assumptions and the different indications and surgical options are examined. PMID- 8237226 TI - Elective cholecystectomy in selected cirrhotic patients. AB - Of the 53 cirrhotic patients with cholelithiasis observed at our Institution from 1978 to 1991, 31 were operated on. Twenty-eight Child-Pugh class A, two class B and one class C patients underwent elective cholecystectomy with (5 cases) or without (26 cases) common bile duct exploration. Among the symptomatic patients, nine (18.7%) were refused for surgery because the risk was estimated to be too high. Symptoms ranged from mild-moderate abdominal pain to typical biliary colic. Acute biliary inflammatory complications as cholecystitis or cholangitis could be detected in the clinical history of 5 patients (16%). Although a common clinical feature, jaundice was directly related to gallbladder or common bile duct stones only in one half of the cases. A total of 18 postoperative non-lethal complications occurred in ten patients (32.2%), with haemorrhage from the gallbladder bed being the most frequent event. Bleeding was associated with increased prothrombin time more than 1.5 seconds above the control (p < 0.01) but severe haemorrhage occurred only when the platelet count was less than 100.000/ml (p < 0.05). Common bile duct explorations increased the risk of bleeding. Two of the 3 class B or C patients developed ascites (p < 0.05). It is concluded that elective cholecystectomy can be performed without mortality in selected and symptomatic patients with adequate hepatic functional reserve. PMID- 8237227 TI - Surgical treatment of hepatic hydatid cysts. AB - Ninety-two surgical procedures were carried out in 82 patients with 92 hepatic hydatid cysts. The most common complication of the hydatid cyst was biliary rupture (17.3%) followed by infection of the cyst cavity (5.4%). Omentoplasty was carried out for uncomplicated cysts (38.0%) with a low morbidity (14.2%) and short hospital stay (mean 12.8 days). External tube drainage was carried out in 30.5% of patients. The morbidity rate was 67.8% and the mean hospital stay was 19.8 days. No single method can be recommended for the treatment of hepatic hydatid cysts but the choice of the surgical method must be made according to the complications of the cyst. Omentoplasty is the procedure of choice for uncomplicated cysts with a low complication rate and relatively short hospital stay. External tube drainage is recommended for infected cysts and a biliary drainage procedure must be added to external tube drainage for cysts with intrabiliary rupture. PMID- 8237228 TI - Asymptomatic gallstones. What to do in patients undergoing colonic surgery for cancer? AB - Twenty-three patients who underwent elective surgery for colorectal cancer in our hospital between 1983 and 1989, underwent concomitant cholecystectomy for asymptomatic gallstones. In order to assess whether additional cholecystectomy increases postoperative morbidity and mortality, a comparison was made between these cases and 23 controlled patients (without gallstones) matched for sex, age (+/- 3 years), Dukes stage and type of primary colonic surgery. The duration of postoperative stay was similar in the two groups (14.1 +/- 4.5 days vs 12.4 +/- 2.3 days). Postoperative complications were more frequent among the case patients than among the controls (34.8% vs 8.7%, p = 0.04), and the same was true for the mortality (8.7% vs 0%). During the same period, another 11 patients with asymptomatic cholelithiasis were operated on for colorectal cancer but in these patients gallstones were left in place. Two of the patients died postoperatively and only one of the remaining 9 (11.2%) had an episode of biliary pain. The results of our study would suggest that in patients undergoing surgery for colorectal cancer the risk entailed in carrying out an additional cholecystectomy for asymptomatic gallstones is greater than the risk of future morbidity caused by gallstones left in place. PMID- 8237229 TI - The Hartmann procedure revisited. AB - The results of 37 consecutive cases of Hartmann's procedure over a 5-year period (1985-1990) were reviewed. The series consisted of 27 emergency procedures (11 diverticular disease, 13 carcinoma, two trauma and one sigmoid volvulus) and ten elective procedures (nine carcinoma and one peridiverticular abscess). The indications for emergency procedures were obstruction and perforation. All patients presented with faecal peritonitis owing to colonic perforation. The mean (range) age was 79.4 (34-90) years. The postoperative mortality rate was 30 per cent overall (11 of 37), 33 per cent (four of 12) in the diverticulitis group, 23 per cent (five of 22) in the carcinoma group, and 100 per cent in the iatrogenic trauma group. Death was mainly due to sepsis (82 per cent). Postoperative complications were mainly wound infections, which occurred in 43 per cent (16 of 37) cases. In 25 per cent of the surviving patients, re-establishment of continuity was performed in three of 17 (18 per cent) of the carcinoma group and three of seven (43 per cent) of the diverticulitis group. No attempt at restoration of intestinal continuity was made in six cases due to medical risk in two, extensive carcinoma in two and local recurrence with metastatic disease in two. Three patients refused all further intervention. There were no postoperative deaths after the restoration of continuity. This series reflects the severity of the pathology in this high risk group of patients. However, the operation can be life-saving for a selected group of patients and offers good palliation for advanced colorectal tumours. PMID- 8237230 TI - Staging of rectal cancer: a prospective study of digital examination and endosonography before and after preoperative radiotherapy. AB - Between January 1988 and January 1992, 34 patients with rectal cancer were evaluated both by clinical examination and endosonography before and after pre operative radiotherapy. Two criteria were correlated with histologic findings: confinement to the rectal wall or spread beyond, the presence of mesorectal lymph node involvement. The 32 patients who underwent endosonography before radiotherapy were staged as: uT2: 4, uT3: 26, uT4: 2 cases. Fifteen days after irradiation, endosonography showed tumour regression in all cases; uT stage was different in 15 patients, uN stage in 4 cases. Comparison of the pre-operative local invasion beyond the rectal wall with postoperative histopathy revealed a correlation with: digital examination after radiotherapy in 20 of the 31 patients with palpable tumours; endosonography before irradiation in 18 of the 32 staged tumours; endosonography after irradiation in 25 of the 32 staged tumours. The presence of mesorectal lymph node involvement determined by histologic examination was correlated with the results of endosonography after radiotherapy for 22 of the 32 staged tumours. Endosonography provides a good assessment of the tumour stage before irradiation. Since radiotherapy alters endosonographic staging of rectal cancer, this staging should be included in survival studies. PMID- 8237231 TI - Primary resection with and without anastomosis for perforation of acute diverticulitis. AB - The results of management of perforated large bowel diverticulitis were retrospectively studied over a 7-year period. 38 patients underwent operation, 20 for generalized peritonitis, 12 for local peritonitis, 5 for colovesical fistula and 1 for colovaginal fistula. The mean age of patients was 63 years (range 30-85 years). Depending on the symptoms, the spreading of the peritonitis and associated cardiovascular and pulmonary disease and diabetes mellitus, 4 types of operation were performed: primary left hemicolectomy and anastomosis with and without defunctioning colostomy, Hartmann procedure, suture and drainage with diverting colostomy. The overall mortality was 10.5%: resection and primary anastomosis entailed 3.8% mortality (1 case), while 3 deaths were observed in the 8 patients group having underwent an Hartmann procedure (37.5%). Drainage and/or diverting colostomy performed in 5 patients entailed no hospital mortality, but was followed by a 80% complication rate, requiring reoperation and several hospital admissions. The low mortality and morbidity rates obtained in the group having primary resection and anastomosis encourage wider application of this operation for perforated acute diverticulitis. Even the Hartmann procedure allows removal of the diseased colon but in a great proportion of cases reconstitution of continuity is not performed; nevertheless staged operation entailing major mortality and morbidity, expose these aged patients to remarkable hazard. Prerequisite of safe primary excision and anastomosis is vigorous intraperitoneal lavage and drainage, by the case associated to on table large bowel irrigation if concomitant obstruction is present. PMID- 8237232 TI - Diffuse cavernous rectal hemangioma--clinical appearance, diagnostic modalities and sphincter saving approach to therapy: report of 2 and a collective review of 79 cases. AB - Two cases of diffuse cavernous haemangioma of the rectum are reported and a thorough review of the world literature with a sum of 79 patients is done. We advocate low anterior resections for the treatment of this disease; sleeve resections are not necessary, since the recurrent bleeding and complication rates are not statistically different from the low anterior resections without mucosal stripping. PMID- 8237233 TI - Surgical management of primary retroperitoneal liposarcomas. AB - During a 21-year period (1970-1991), 7 patients with primary retroperitoneal liposarcomas were treated surgically in our department. All the tumours were excised completely, and in 2 patients an additional nephrectomy was performed. Postoperatively 4 patients had adjuvant radiotherapy. Two other patients received adjuvant chemotherapy. Both of them had to be reoperated, because of a local recurrence 12 and 9 months after their first operation. There was one death on the first postoperative day, and 4 patients died 1 1/2-2 years after the initial operation. Two patients are still alive 34 and 22 months after their first operation with recurrent disease. Adjuvant therapy proved to be ineffective on the survival of our patients. The prognosis of retroperitoneal liposarcomas continues to be poor and the primary treatment has to be aggressive with complete resection of the tumor. PMID- 8237234 TI - Virilizing adrenal ganglioneuroma containing Leydig cells. AB - An adrenal medullary ganglioneuroma containing Leydig cells and revealed by a virilizing syndrome in a 56-year old woman is presented. The syndrome, associating with masculinization an elevated serum testosterone level and a normal urinary 17 ketosteroids, is uncommon in adrenal tumours. The tumour was located by computed tomographic (CT) scan and treated by right adrenalectomy. Microscopic examination showed typical features of an adrenal medullary ganglioneuroma containing Leydig cells with Reinke crystalloids. Positive immunohistochemical study confirmed the testosterone secreting nature of the cells. The presence of Leydig cells in the adrenal gland is discussed according embryogenic studies: 1. the common origin of the gonad and the adrenal cortex from the coelomic epithelium, 2. the vicinity of adrenocortical gland and gonad during embryogenesis, 3. the thecal metaplasia of mesenchymal cells, 4. the development of Leydig cells from Schwann cells. Fourth case published in the world literature. PMID- 8237235 TI - Intraoperative real-time ultrasonography in the microsurgical removal of subcortical or deep-seated brain tumors. AB - Seventy-three patients with subcortical or deep-seated brain tumours have been operated on with the aid of intraoperative ultrasonography. Forty-one had metastases and thirty-two primitive neuroepithelial neoplasms. In all cases ultrasonography allowed an easy and precise localization of the tumours. The removal was gross total in 45 cases (61.6 per cent), subtotal in 17 (23.3 per cent) and partial in 11 (15.1 per cent). Preservation or improvement of the Karnofsky Performance Status was obtained in 63 patients (86.3 per cent), with a morbidity and mortality rate of 11 and 2.7 per cent respectively. Our results indicate that ultrasound guided microsurgery represents a handy, reliable and relatively low risk procedure for the treatment of intra-axial brain tumours. PMID- 8237236 TI - Transpedicular fixation in the treatment of various spinal disorders. AB - 36 patients with 157 inserted pedicular screws are presented. The patients are evaluated in 5 different groups; group I: fractures, group II: spinal tuberculosis, group III: spinal tumours, group IV: spondylolisthesis and group V: scoliosis. The mean age and the mean follow-up of the groups are respectively 42 years and 10.8 months. Cotrel-Dubousset instrumentation (CD), Alici Instrumentation, Fixateur Interne (Dick) or Steffee plate are used to stabilize the spine. The low complication rate of pedicular screw insertion is related to the used technique. PMID- 8237237 TI - Spontaneous splenic rupture in infectious mononucleosis. AB - A young man with an infectious mononucleosis underwent an emergency splenectomy for a spontaneous splenic rupture. This is a rather rare complication of mononucleosis caused on the one hand by a change in the histological structure of the spleen and on the other hand by a splenomegaly. The prognosis is favourable when the diagnosis is made on time and the correct treatment has been started immediately. With the exception of those cases which are haemodynamically stable (the lesion is limited and corresponds to a subcapsular haematoma), splenectomy remains the treatment of choice. Conservatively treated patients must be closely followed and at the first sign of destabilization, surgical intervention is required as soon as possible. PMID- 8237238 TI - Atopic diseases in Norwegian Lapps. AB - The occurrence of atopic diseases in Lapps was estimated by reviewing medical records in the local health centre of Kautokeino. Atopic diseases were found to be present or to have occurred in 306 (10.4%) of the 2,950 individuals reviewed, with no difference between males (10.6%) and females (10.2%). Atopic diseases were detected more frequently in those under 20 years (19.4%) than in those 20 years or older (5.5%). On average, asthma occurred more frequently in males (4.2%) than in females (2.9%), even more so in the younger age group (males = 7.5%, females = 2.5%). Similarly, the occurrence of allergic rhinoconjunctivitis was remarkably low, but found in more males (2.3%) than in females (1.9%) with the frequency increasing in those under 20 years (males = 4.7%, females = 2.9%). A peak prevalence was observed in the age group 10-19 years (6.1%). On the other hand, atopic urticaria was found more frequently in females (1.0%) than in males (0.3%), whereas atopic dermatitis occurred with the same frequency in females (6.4%) and males (6.1%) followed by the frequency in those under 20 years increasing to 13.9% and 13.5%, respectively. PMID- 8237239 TI - Declining trends in some sexually transmitted diseases in Norway between 1975 and 1991 with special reference to a Lapp population. AB - Trends of N. gonorrhoeae (1975-1991) and C. trachomatis (1988-1991) infections in Norway were analysed by the National Institute of Public Health (SIFF) using data reported by physicians. The validity of the reporting system was evaluated by comparing reported data from the three northernmost counties and particularly the Kautokeino community to SIFF with confirmed positive specimens analysed by the two microbiological laboratories serving this area. The incidence of gonorrhoea in Norway has declined continuously from 300 cases per 100,000 annually in 1976 to 15 in 1991, whereas C. trachomatis infections with annual rates of 300 cases per 100,000 during the last 4 years has shown no significant decrease. The incidence of gonorrhoea in the county of Finnmark has shown the same declining trend as for the rest of the country, but is still four times higher than the national average. C. trachomatis infections show no obvious declining tendency in any part of the country and is three to four times higher in Finnmark than the average for the country. Kautokeino experienced a dramatic decrease in gonococcal infections from more than 1000 cases per 100,000 in 1976 to almost eradication in 1991, whereas C. trachomatis infections show a four times higher annual incidence than the national average. Although gonorrhoea is almost eradicated in Norway, C. trachomatis infections remain an important public health problem. This study indicates that young, sexually active individuals need to be targeted for effective health education in order to modify behaviour patterns which put them at risk of acquiring STDs. PMID- 8237240 TI - Prevalence of dermato-venereological diseases in a Norwegian Lapp population. Introduction. PMID- 8237241 TI - Prevalence of psoriasis in a Norwegian Lapp population. AB - The occurrence of psoriasis in a Norwegian Lapp population was estimated by reviewing medical records in the local health centre of Kautokeino. Altogether 40 cases of psoriasis were registered in the study population comprising 99.6% of the total population of 2,963 individuals. Thirty-five cases of psoriasis belonged to the Lappish population of 2,508 people, giving a prevalence of 1.4%, with no difference between males and females. Five cases of psoriasis were found in the non-Lappish population of 442 individuals which gives a prevalence of 1.1% with no sex differences. A peak prevalence among Lapps was observed in the age groups 20-39 years (2.7%), equal for both sexes and in females aged 50-59 years (3.2%), whereas a lower prevalence of 0.6% was found in the age group 40-49 years. A seronegative psoriatic arthritis was recorded in 6 of the 35 Lappish compared to none of the non-Lappish cases. A familial association was confirmed in 11 of the 35 Lappish cases, in none of the non-Lappish. PMID- 8237242 TI - Adrenal imaging in Cushing's syndrome: morphology or function? PMID- 8237243 TI - Unusual presentation of a TSH-secreting pituitary adenoma. PMID- 8237244 TI - Hyperfunctioning unilateral adrenal macronodule in three patients with Cushing's disease: hormonal and imaging characterization. AB - We aimed to investigate the dynamics of adrenocorticotropin (ACTH) and cortisol secretion in pituitary-dependent Cushing's syndrome with bilateral macronodular adrenal hyperplasia presenting as a single adrenal macronodule, and to determine the imaging characteristics of this syndrome. Three female patients were studied. Plasma ACTH and serum cortisol secretion were studied by determining their rhythmicity and pulsatility and their responses to the administration of ovine corticotropin-releasing factor, thyrotropin-releasing hormone, metyrapone, tetracosactrin, insulin and dexamethasone. Techniques used to localize the anatomical lesion were bilateral simultaneous inferior petrosal sinus sampling, magnetic resonance examination of the pituitary, computed tomography (CT) scanning and [75Se]cholesterol scintigraphy of the adrenal glands. Plasma ACTH and serum cortisol levels were measured using a commercial radioimmunoassay and an immunoradiometric assay. The ACTH and cortisol pulse number and amplitude were calculated using established computer software. In all three patients ACTH and cortisol secretory dynamics fulfilled the requirements for diagnosis of pituitary dependent Cushing's syndrome. A close relationship between ACTH and cortisol pulses also favored a pituitary dependency. Study of the amplitude of cortisol pulses classified two patients in the group of hypopulsatile Cushing's disease. Adrenal CT scanning demonstrated the presence of a large single nodule. [75Se]Cholesterol scintigraphy showed bilateral radionuclide uptake, although mostly localized over the adrenal nodule. All patients underwent successful trans sphenoidal hypophysectomy. Over a period of 1 year, a slow shrinkage of the adrenal nodule was observed in two patients, while no change in volume was observed in one patient.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237245 TI - The prevalence of radiographic osteoarthritis is low in elderly patients with growth hormone deficiency. AB - The prevalence of radiographic osteoarthritis in hand and knee joints was studied in elderly patients with acromegaly and growth hormone deficiency, respectively, and compared with a normal population of elderly people. There were no major differences in the prevalence of osteoarthritis between the acromegalics and the normal population, but the patients with growth hormone deficiency had significantly (p < 0.001) less osteoarthritis than the normal population. The lack of differences between the acromegalics and the normal population could be an effect of the age interval studied in which the prevalence of osteoarthritis is high. The low prevalence of osteoarthritis in patients with growth hormone deficiency suggests that growth hormone is an important factor in the development of osteoarthritis. PMID- 8237246 TI - Analysis of the Gs alpha gene in growth hormone-secreting pituitary adenomas by the polymerase chain reaction-direct sequencing method using paraffin-embedded tissues. AB - We investigated the prevalence of Gs alpha gene mutations in growth hormone (GH) secreting pituitary adenomas from Japanese patients with acromegaly. Forty-five GH-secreting adenomas were examined for the presence of point mutations in codons 201 or 227 of the Gs alpha gene using the polymerase chain reaction-direct sequencing method and deoxyribonucleic acid extracted from paraffin-embedded tumor specimens. Mutation of codon 227 of the Gs alpha gene was not observed in any of the tumors, but a mis-sense mutation of codon 201 was identified in two tumors (4.4%). One lesion was a densely granulated GH cell adenoma in a patient with adenomatous goiter and breast cancer. The other was a mixed GH cell prolactin cell adenoma in a patient with multiple endocrine neoplasia type 1 associated with parathyroid hyperplasia and a pancreatic islet cell tumor. The Gs alpha gene detected in parathyroid tissue and pancreatic tumor tissue was of the wild type in this second patient, and the mutation was specific to the pituitary tumor. These results suggest that point mutations of codons 201 or 227 of the Gs alpha gene may not be important mediators of oncogenesis for GH-secreting pituitary adenomas in Japan. PMID- 8237247 TI - Hormonal status and clinical relevance of hirsutism in elderly women. AB - Hirsutism is a common condition of elderly women, but its aethiopathogeny and its clinical implications remain unclear. We therefore studied circulating androgen concentrations in elderly women. In addition, this study aims to define a possible relationship between hirsutism and anthropometric determinations, bone mass and serum lipids. Androgen levels were determined at basal state, after adrenocorticotrophin (ACTH) stimulation and after dexamethasone administration in 10 hirsute elderly women and compared to 10 age-matched non-hirsute women. Anthropometric determinations included measurements of skinfold thickness and body mass index. Spinal bone mass density was assessed using dual photon absorptiometry. Hirsute women presented significantly higher levels of testosterone than controls (1.49 +/- 0.38 vs 0.59 +/- 0.05 nmol/l, mean +/- SEM; p < 0.05) and dihydrotestosterone (0.54 +/- 0.07 vs 0.32 +/- 0.03; p < 0.02). 17 hydroxyprogesterone levels after ACTH stimulation tended to be higher in hirsute women than in controls. No differences were observed between the two groups in serum oestrogen concentrations, plasma lipid pattern or bone mineral density. Hirsute women had a lower body mass index and lower calculated percentage body fat than the control group. We conclude that: (i) hirsutism of elderly women is associated with increased androgen levels, probably from adrenal origin; (ii) in some cases, enhanced response in 17-hydroxyprogesterone after ACTH stimulation suggests a partial adrenal 21-hydroxylase deficiency; (iii) hirsute women present anthropometric characteristics compatible with the known anabolic effect of androgens on fat-free mass. PMID- 8237248 TI - Peripheral vein, ovarian vein and ovarian tissue levels of inhibin in a postmenopausal patient with a granulosa cell tumour. AB - In a patient with a granulosa cell tumour, stage 1a1, peripheral and ovarian vein blood samples were drawn before, during and after bilateral oophorectomy. Before operation inhibin and follicle-stimulating hormone levels were in the premenopausal range, whereas the peripheral level of oestradiol and luteinizing hormone were in the normal postmenopausal range. In the ovarian vein at the side of the tumour, inhibin and oestradiol levels were elevated, whereas in the contralateral ovarian vein the concentrations of inhibin and oestradiol were in the same range as in the peripheral vein. Immunoreactive inhibin levels in the homogenized tumour were 23 times higher than in the contralateral ovary, whereas inhibin bioactivities in the same samples amounted to 126 and 14 U/mg protein, respectively. After removal of the tumour, peripheral serum follicle-stimulating hormone and inhibin levels were in the normal postmenopausal range again. We conclude that inhibin can have a role as a marker for granulosa cell tumours. PMID- 8237249 TI - Long-term endocrine effects of administration of either a non-steroidal antiandrogen or a luteinizing hormone-releasing hormone agonist in men with prostate cancer. AB - The claimed ability of non-steroidal antiandrogens to preserve libido and sexual potency is sought as a potential improvement in the palliative management of prostate cancer. A critical issue for the clinical use of these compounds is, however, the reported evidence in the rat of an excessive increase in testosterone concentrations as a consequence of the androgen negative feedback interruption. On the other hand, the recovery of testicular function after long term inhibition by luteinizing hormone-releasing hormone (LHRH) analogs is also an important concern in view of the proposed use of these compounds for the treatment of several non-malignant conditions. We addressed these issues by studying the long-term endocrine effects induced by the administration of either the non-steroidal antiandrogen nilutamide or the depot preparation of D-Trp6-LHRH in men with prostate cancer. Treatment with the antiandrogen induced a marked increase in gonadotropin levels, LH concentrations rising from a mean (SEM) of 17.5 +/- 1.6 to a maximum of 56.6 +/- 6.9 kU/l (p < 0.001), while mean testosterone and 17 beta estradiol-concentrations rose only by about 50% and 70% over pretreatment values, testosterone levels reaching a plateau after 1 month of treatment. In the subjects treated with the LHRH agonist, 6 months after discontinuation of long-term administration the mean (+/- SEM) LH had risen to 36.9 +/- 6.8 IU/l while mean testosterone levels were still as low as 1.7 +/- 0.7 and rose only to a maximum of 4.2 +/- 1 nmol/l after high-dose human chorionic gonadotropin loadings.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237250 TI - Pentoxifylline inhibits the proliferation and glycosaminoglycan synthesis of cultured fibroblasts derived from patients with Graves' ophthalmopathy and pretibial myxoedema. AB - Excessive amounts of glycosaminoglycans accumulate in the extraocular muscles of patients with Graves' ophthalmopathy and in the affected skin of patients with pretibial myxoedema. It is widely accepted that fibroblasts are the sources of glycosaminoglycan synthesis. Pentoxifylline, an analogue of the methylxanthine theobromine, inhibits the proliferation and certain biosynthetic activities of fibroblasts derived from normal human skin and from skin of patients with some fibrotic disorders. Our objective was to determine whether pentoxifylline has similar effects on fibroblasts derived from patients with Graves' ophthalmopathy and pretibial myxoedema and could serve as a candidate for the treatment of these manifestations. Fibroblasts from the extraocular muscles of two patients with Graves' ophthalmopathy and normal extraocular muscles of two subjects with strabismus, as well as the affected skin of two patients with pretibial myxoedema were cultured in vitro in the presence and absence of pentoxifylline to assay its effect on the proliferation of fibroblasts and their production of glycosaminoglycans. In subconfluent fibroblast cultures, pentoxifylline treatment caused a dose-dependent inhibition of serum-driven fibroblast proliferation. In confluent fibroblast cultures both in the presence and absence of serum, exposure to pentoxifylline similarly resulted in a dose-dependent inhibition of glycosaminoglycan synthesis for all these different kinds of fibroblasts. These findings may form the rationale for a clinical trial using pentoxifylline for the treatment of Graves' ophthalmopathy and pretibial myxoedema. PMID- 8237251 TI - Thyroid peroxidase: kinetics, pH optima and substrate dependency. AB - The oxidation of iodide, guaiacol and 2,2'-azino-di[3-ethyl-benzthiazoline-(6) sulphonic acid] and the iodination of tyrosyl residues in bovine serum albumin, catalysed by partly purified thyroid peroxidase, were studied. The enzyme showed pH optima with all electron donors. With the exception of guaiacol, the position of the pH optima depended upon both the electron donor and hydrogen peroxide concentrations. With increased hydrogen peroxide concentrations the optima shifted to lower pH, and with increased iodide concentration to higher pH. For monoiodotyrosine (MIT) formation in bovine serum albumin the position of the pH optimum was also dependent on the hydrogen peroxide concentrations. The position of the pH optimum of the oxidation of guaiacol was pH 9 and independent of substrate and hydrogen peroxide concentrations. It is obvious from these findings that iodination reactions must be studied under well-defined conditions. PMID- 8237252 TI - Parathyroid hormone-related peptide and milking-induced phosphaturia in dairy cows. AB - Five intact and four thyroparathyroidectomized (TPTX) lactating dairy cows were fitted with polyvinyl catheters implanted into the left carotid artery and the left mammary vein (for blood sampling) and into the right external jugular vein (for injections), and with a Foley catheter inserted into the urinary bladder. In these nine cows, daily morning and evening milking induced a prompt and transient rise in mammary venous blood plasma parathyroid hormone-related peptide (PTHrP)(1 34) concentrations, followed by an increase in both urinary phosphate concentration and phosphate renal clearance. Such a milking-induced phosphaturia was not observed when (tyr)34-bPTH(7-34)-NH2 (5 pmol/kg body wt) was infused during milking into the TPTX cows. These results indicate that PTHrP released systematically by the mammary gland during milking might be responsible for the observed phosphaturia. PMID- 8237253 TI - Beta-adrenergic receptors, voltage-operated Ca(2+)-channels, nuclear triiodothyronine receptors and triiodothyronine concentration in pig myocardium after long-term low-dose amiodarone treatment. AB - Similar features during chronic amiodarone treatment and hypothyroidism suggest that amiodarone induces a state of "triiodothyronine (T3)-resistance" or "cardiac hypothyroidism", which may predispose the heart to pump failure under conditions with severe strain, such as recovery after cardiac surgery. Disagreements exists as to how amiodarone, and possibly its main metabolite desethylamiodarone, act upon the various receptor systems in the heart. The aim of the present study was to elucidate whether chronic amiodarone treatment leads to a functional reduction in the number of myocardial nuclear T3 receptors, the myocardial tissue T3 concentration and the number of beta-receptors and voltage-operated Ca(2+) channels. Finally, special attention was drawn to any changes that could contribute to explain previous reports on reduced haemodynamic reserve in animals exposed to severe cardiac strain, such as cardiac surgery. Pigs (72 +/- 2 kg) were assigned randomly to amiodarone treatment (20 mg.kg-1.day-1 for 30 +/- 1 days, N = 8); controls received no medical treatment (N = 6). The left ventricle was evaluated for beta-adrenergic receptors, voltage-operated Ca(2+)-channels, T3 nuclear receptors and tissue T3 concentration. Maximum binding capacity for beta receptors and Ca(2+)-channels was reduced in amiodarone-treated pigs (by 38%, p < 0.05, and by 52%, p < 0.01) and correlated with tissue drug concentrations for both receptor types (p < 0.05). No changes were observed concerning nuclear T3 receptors. In vitro competition studies revealed that amiodarone, but not desethylamiodarone, possessed binding properties to Ca(2+)-channels, whereas neither of the compounds bound to beta-receptors. Desethylamiodarone, but not amiodarone, competitively inhibited T3 binding to its nuclear receptors. Myocardial T3 was undetectable (< 0.05 nmol/kg wet wt) in amiodarone-treated pigs. From our observations we suggest that the active metabolite desethylamiodarone, rather than the parent drug, is mainly responsible for the observed local hypothyroid-like effects during amiodarone treatment. The observed changes after treatment with low-dose amiodarone in pigs are likely to have biological implications. Functionally, the changes may imply reduced cardiac reserve during conditions of extraordinary strain. PMID- 8237254 TI - Interaction of unsaturated fatty acids with rat liver glucocorticoid receptors: studies to localize the site of interaction. AB - Polyunsaturated fatty acids have been shown to decrease the binding of [3H]dexamethasone to rat liver glucocorticoid receptors by mixed non-competitive inhibition, suggesting that these fatty acids interact at a site on the receptor different from the hormone binding site. The present study was undertaken to localize the site of interaction of polyunsaturated fatty acids on the receptor by comparing the differential effects of docosahexaenoic acid (a 22-carbon polyunsaturated fatty acid of the series n-3) on antagonist (RU486) and agonist binding, by covalent cross-linking of the hsp 90 and other proteins to the receptor to attempt to mask the site of interaction, by limited trypsinization to cleave the site and by using antibodies against specific epitopes to prevent fatty acid access by steric hindrance. Binding [3H]RU486 was not inhibited by docosahexaenoic acid at a concentration (60 mumol/l) that increases the dissociation constant of [3H]dexamethasone eightfold. Covalent stabilization of the hetero-oligomeric glucocorticoid receptor structure did not keep the fatty acid from inhibiting [3H]dexamethasone binding. The binding to the receptor of monoclonal and polyclonal antibodies against different domains of the receptor did not sterically hinder the fatty acid interaction with the receptor. After limited trypsinization of the receptor, the fatty acid still increased the dissociation rate constant of [3H]dexamethasone binding, indicating that the site of interaction of polyunsaturated fatty acids is on a fragment of the receptor containing the hormone-binding domain and some sequences C-terminal of the DNA binding domain. PMID- 8237255 TI - Concentrations, metabolic clearance rates, production rates and plasma binding of cortisol in Antarctic phocid seals. AB - We have reported previously that plasma of the Weddell seal, a member of the phocid family, contains a very high concentration of cortisol. The present study was undertaken to determine whether high cortisol levels were common to seals in the Antarctic environment, or to other phocidae, and to determine the mechanism of the hypercortisolaemia. High levels of cortisol (0.82-2.38 mumol/l) were found in 4 phocidae (Weddell, crabeater, leopard and Southern elephant seals), whereas levels in a member of the otariid family (Antarctic fur seal) were similar to human values. Metabolic clearance rates (MCR) and production rates (PR) of cortisol were determined in the field in Weddell (N = 1), crabeater (N = 3) and leopard (N = 3) seals following bolus injections of [3H] cortisol. The MCR and PR did not differ between the three phocids, but whereas the MCR of 410-590 1/day was twice that of human values, the PR of 460-1180 mumol.m-2 x d-1 was up to 40 fold greater. The binding capacity of corticosteroid-binding globulin (CBG) was equal to or greater than the plasma concentrations of cortisol, resulting in relatively low concentrations of free cortisol. We conclude that hypercortisolaemia is maintained in phocid seals mainly by a high production rate -the highest (corrected for surface area) reported in any species. The relatively low cortisol levels in otariid seals studied in the same environment suggest that the high PR in phocidae is unrelated to the harsh climatic conditions, but may be part of their adaptation for diving to extreme depths.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237256 TI - Insulin resistance caused by amylin in conscious rats is independent of induced hypocalcemia and fades during long-term exposure. AB - To compare the effect of short- vs long-term amylin infusion on insulin sensitivity, glucose tolerance and serum calcemia, euglycemic-hyperinsulinemic clamp (26 pmol.kg-1 x min-1) and glucose tolerance tests (2.4 mmol/kg over 30 min) were performed in lean Zucker rats. Three infusion protocols were employed: control group: 24 h of i.v. saline; short-term amylin exposure: 22 h of i.v. saline followed by 2 h of i.v. amylin (20 micrograms/h); long-term amylin exposure: 24 h of iv amylin (20 micrograms/h). Insulin resistance was induced by short-term amylin infusion during euglycemic clamping, as shown by a 41% decrease in space-corrected glucose infusion rates (mumol.kg-1 x min-1; control group, 106.0 +/- 15.0; short-term i.v. amylin, 62.7 +/- 15.0; p < 0.005). After long term amylin exposure, insulin sensitivity was identical to control values (109.9 +/- 6.7). This fading action of amylin was confirmed by data from the glucose tolerance test, demonstrating glucose intolerance after short- but not after long term amylin exposure. Serum calcium concentration decreased during short-term (2 h) amylin infusion (from 2.52 +/- 0.15 to 2.09 +/- 0.12 mmol/l; p < 0.01) and hypocalcemia of a similar extent also was present after 22 h and 24 h of amylin exposure (2.10 +/- 0.09 and 2.04 +/- 0.14 mmol/l, respectively). The data demonstrate that short-term amylin infusion induces insulin resistance and glucose intolerance, both of which vanish during long-term (> 22 h) amylin exposure, being apparently independent of induced hypocalcemia. PMID- 8237257 TI - Amount of G-protein alpha-subunit in rat white adipocytes: lack of difference between subcutaneous and visceral fat. AB - It has been the purpose of this study to examine possible differences in the amount of stimulatory (Gs) and inhibitory (Gi) G-protein alpha-subunits (measured with a quantitative enzyme-linked immunosorbent assay in fat cell membrane preparation) between subcutaneous and intra-abdominal regions in rats. The lipolytic response to isoproterenol and the number of beta-adrenergic binding sites were also examined. These parameters were all evaluated simultaneously in subcutaneous (inguinal), epididymal and perirenal fat samples collected from six male Sprague-Dawley rats. The membrane contents of the Gs and Gi alpha-subunits were similar in the three depots. Moreover, no difference was found among the different regions with regard to isoproterenol-stimulated glycerol release and beta-adrenoceptor number, expressed per cell number. In conclusion, the present study shows for the first time in rats that the abundance of inhibitory and stimulatory G-protein alpha-subunits is similar in subcutaneous and in visceral adipocytes. Moreover, the number of beta-adrenoceptors and the lipolytic response to isoproterenol do not show significant variations with the anatomical site. As the present results are apparently in contrast with those obtained previously in human adipocytes, there is a possibility that the different results observed in rat and in human fat cells could be explained by species differences. PMID- 8237258 TI - Endocrine profile of catch-up growth in the cynomolgus monkey. AB - We studied catch-up growth following withdrawal of glucocorticoid administration in seven intact prepubertal cynomolgus monkeys. To reduce stress during blood sampling, a vascular access port was implanted subcutaneously in each animal for the duration of the study. After a baseline observation period of 50 weeks, the monkeys received injections of dexamethasone at a dose of 100 micrograms.kg-1.day 1 for 15 weeks. Growth velocity was monitored every 3 weeks by measuring lower leg length and body weight. Spontaneous serum growth hormone (GH) concentrations and GH levels after insulin and L-dopa stimulation, as well as serum insulin-like growth factor I (IGF-I) and plasma thyrotropin, triiodothyronine and thyroxine, were measured during the study. Differences between animals were analyzed by repeated measures analysis of variance and Student's paired t-test. Mean +/- SEM growth velocity (mm/3 weeks) decreased from 0.90 +/- 0.08 during the baseline period to 0.29 +/- 0.07 (p < 0.001) during the period of growth retardation, and increased to 1.23 +/- 0.2 (p < 0.001) during the period of catch-up growth. Spontaneous GH and peak GH levels following insulin did not show any significant variation during the study. Peak GH during the L-dopa test decreased from 15.4 +/ 3.2 micrograms/l during the baseline period to 6.2 +/- 2.4 micrograms/l during the period of growth retardation (p < 0.05), and increased to 23.0 +/- 5.9 micrograms/l during the period of catch-up growth (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237259 TI - [Effectiveness and safety of 150 mg vs 300 mg ranitidine twice a day in duodenal ulcer]. AB - 394 patients with endoscopically diagnosed duodenal ulcer were randomly allocated to treatment with ranitidine 150 bid o ranitidine 300 mg bid in a prospective double-blind multicenter trial conducted in seven LatinoAmerican countries. Endoscopy at 4 weeks showed complete ulcer healing en 171 of 196 patients (87.2%) treated with ranitidine 150 mg bid and 178 of 198 (89.9%) treated with ranitidine 300 mg bid. Both treatment regimens were equally effective at rapidly reducing the incidence of ulcer-related symptoms. It is possible that higher dosage regimen of ranitidine would be useful in patients with more severe duodenal ulcer disease. PMID- 8237260 TI - [Correlation of IgA class antigliadin and antiendomysial antibodies (IgA-AGA-IgA EMA) with intestinal histology in celiac disease]. AB - The aim of this work was to establish the diagnostic and follow up value of IgA class antiendomysium (IgA-EmA) and IgA-class antigliadin (IgA-AGA) antibodies in celiac disease. Correlation with the intestinal histology at the different stages of the disease was evaluated, as well as its therapeutic monitoring ability. Fifty six children, twenty seven girls and twenty nine boys, aged six months to twelve years old, were studied. Thirty nine celiac children were al different diagnostic stages of the disease. Seventeen children with malabsorption symptoms and with normal intestinal histology were used as controls. Sixty blood samples were obtained simultaneously with the small intestinal biopsy. IgA-AGA (ELISA method) and IgA-EmA (immunofluorescent test performed over lower third Rhesus monkey esophagus) were determined in every blood sample. In 34 serum samples from patients with total or subtotal villous atrophy, two were negative for IgA-AGA and only one was negative for IgA-EmA. In 26 samples from patients with normal intestinal histology, two were positive for IgA-AGA and four were positive for IgA-EmA. The results for IgA-EmA had sensitivity 97%, specificity 84.6%, positive predictive value 89.2% and negative predictive value 95%. In the case of IgA-AGA were: sensitivity 94.1%, specificity 92.3%, positive predictive value 94.1%, negative predictive value 92.3%. IgA-AGA and IgA-EmA showed a high correlation with intestinal histology and are in combination powerful tools for the diagnosis and follow up of celiac patients. Besides, they provide a useful aid in the indication of a jejunal biopsy and in close monitoring of the dietary treatment compliance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237261 TI - [Endoscopic sphincterotomy. Complications and follow-up]. AB - Between April 1981 and April 1991, there were 547 EPT performed, seventy-five of which had in situ gall bladder. They were performed in two groups, who were similar from the technical-anatomical point of view, and sociocultural background. The early morbidity showed 31 complications (5.6%). The most frequent ones were: perforation in 8 cases and haemorrhage in 5. Medical treatment, which included the endoscopic technique in 20 cases (3.7%) and surgical procedures in 11 cases (2.0%) was used. The mortality rate was 1.8% (10 cases), perforation (4) and cholecystitis (3) being the main causes. There were three deaths (0.5%) not related to the method. The morbi-mortality was not significantly increased by the complementary methods. After analysing the complications related to the cause of EPT performance, it was observed that they were more frequently produced in choledochal stones: 20 (3.6%), followed by choledochal and gall bladder stones: 7 (1.3%). However the mortality was 0.7% in the former and 0.5% in the latter. The morbi-mortality of the last two years is compared to that of the preceding ones. The follow-up was done in 114 (20.8%) of the cases with a marked difference between the high sociocultural group (76 cases) and the low one (36). Eighteen pathological cases were found (3.3%). The treatment of each complication in long term is shown. PMID- 8237262 TI - Chronic non specific gastritis: problems today. PMID- 8237263 TI - [Prophylactic antibiotics in acute necrotic pancreatitis]. PMID- 8237264 TI - [Functions of the colon]. PMID- 8237265 TI - Gastric chief cell mass in duodenal ulcer. AB - Chief cell mass and serum Pepsinogen I were calculated in 25 duodenal ulcer patients and in 40 healthy controls. Comparisons were also made with parietal cell mass and the stimulated acid secretion. Chief cell mass was expressed bay a Zymogenous Index (ZI) obtained by multiplying the number of chief cells per mm2 for the thickness of the glandular layer. The analysis of the data shows a significant decrease of ZI in duodenal ulcer patients in comparison with controls. The grouping of the subjects according to the age (over and under fifty years), confirms that ZI decreases significantly in duodenal ulcer groups in relation with the respective groups of control. In case of duodenal ulcer with fundic superficial gastritis no behavioural changes of ZI are observed in comparison to those affected by duodenal ulcer with normal fundic mucosa. The serum Pepsinogen I significantly increases in duodenal ulcer patients in comparison to the controls, either in the subjects over and under fifty. In the group of duodenal ulcer patients with superficial fundic gastritis a significant increase of serum Pepsinogen I can be pointed out in comparison to the ulcer patients with normal fundic mucosa. The grouping of the patients on the basis of hyperparietalism and normoparietalism has shown in the former group hypozymogenism and in the latter normozymogenism. Finally, no correlation has emerged between Zymogenous Index and serum Pepsinogen I. PMID- 8237266 TI - [Irritable intestine: a personal approach]. PMID- 8237267 TI - Quantitative and qualitative characteristics of colony-forming unit-erythroid colonies in myelodysplastic syndrome patients. AB - The microcytofluorometrical method was applied to determine the relative hemoglobin (Hb) content in the bone marrow colony-forming unit-erythroid (CFU-E) colonies from 6 patients with myelodysplastic syndromes (MDS) and 10 healthy subjects. This method relies on a photochemical reaction, by which intracellular Hb is converted into fluorescent porphyrin using a 0.2 M mercaptoethylamine solution (an SH donor) and violet light (lambda = 405 nm). The relative Hb content was determined as a function of the intensity of emitted porphyrin fluorescence. The number of colonies identified by porphyrin fluorescence was smaller in MDS patients than in normal subjects. The relative Hb content was also lower in MDS patients than in normal subjects. In addition, the coefficient of variation of the relative Hb content in the CFU-E colonies was larger in MDS patients than in normal subjects. These findings suggest that colonies with low relative Hb content undergo impaired erythropoiesis and that the CFU-E colonies undergoing the impaired erythropoiesis are mixed with CFU-E colonies showing normal erythropoiesis in the bone marrow of MDS patients. PMID- 8237268 TI - Variations in erythropoiesis and serum ferritin during erythropoietin therapy for anaemia of end-stage renal disease. AB - In order to study the relationship between erythropoiesis and serum ferritin (SF) during erythropoietin (rHuEPO) therapy in the anaemia of end-stage renal disease (ESRD), 19 patients were followed without iron supplementation and at a fixed dose of the drug (40 U/kg). Twelve patients failed to attain the target haemoglobin (Hb) value, 7 of whom due to the appearance of iron deficiency. Erythropoiesis, as measured by the serum transferrin receptor concentration, increased from 12 to 120% of the basal value. This increment was not constantly associated with a proportional rise of Hb or reticulocyte count. SF decreased exponentially from a median value of 221 micrograms/dl (range 42-470) to a median value of 54 micrograms/dl (range 20-172). Halving of the basal SF value (SF-T50) was reached at the 18th-95th day of therapy (median = 43), representing a iron shift of 3.4-11.6 mg/day (median = 5.4). SF-T50 was not correlated with the Hb increase, but with that of erythropoiesis (r = 0.78; p = 0.003). The minimum SF (MSF) value attained was not correlated with the appearance of iron deficiency. The conclusion is that the rate of SF decrease during rHuEPO in ESRD is a reliable measure of iron mobilisation for erythropoiesis, but not for haematologic response. The MSF value reached during therapy is not representative of available iron for erythropoiesis. PMID- 8237269 TI - Characterization of myeloid or lymphoid acute leukemia by a chemiluminescence assay. Comparison with immunocytochemistry using an antimyeloperoxidase antibody. AB - A simple and sensitive chemiluminescence assay for the demonstration of the activity of intracellular myeloperoxidase (MPO) is described, which is useful for the distinction between myeloid and lymphoid commitment in blasts from acute leukemia patients. When the cut-off point was settled at 13 mV of chemiluminescence all cases of acute myeloid leukemia (AML) were distinguished from those of acute lymphoid leukemia. In addition, this technique was able to demonstrate MPO activity in AML poorly differentiated (FAB-M0) which usually does not stain for MPO in classical cytochemistry preparations and could be negative also by immunocytochemistry with anti-MPO monoclonal antibody. Therefore the method here described presented a higher sensitivity than the immunocytochemistry procedure with anti-MPO. PMID- 8237270 TI - Low-grade haemolysis and assessment of iron status during the steady state in G6PD-deficient subjects. AB - We evaluated the iron status of 50 Sicilian patients with G6PD deficiency under steady-state conditions and compared our results with those for 50 control patients. We studied haemolysis and iron indices to evaluate the iron balance. These patients could be considered to be at risk of iron overload as a result of increased bone marrow activity. Reticulocytosis and macrocytosis with reduced levels of haptoglobin were found in the G6PD-deficient subjects, both of which are evidence of a moderate haemolysis. Iron status within the normal range, without iron overload or iron deficiency, was found. PMID- 8237271 TI - Endothelial activation antigens in pulmonary leukostasis in leukemia. AB - Adhesion between leukemic cells and the vascular endothelium has been suggested to play a role in the development of leukostasis in myelocytic leukemia. To define the role of adhesion molecules on the surface of endothelial cells in leukostasis, we used immunohistochemistry to study the expression of endothelial leukocyte adhesion molecule-1 (ELAM-1), vascular cell adhesion molecule-1 (VCAM 1), and intercellular adhesion molecule-1 (ICAM-1) in lung tissue of 4 patients with pulmonary leukostasis. Lung tissue of 2 patients with myelocytic leukemia without leukostasis and 4 patients with irrelevant nonpulmonary disease was used as a negative control. Positive control tissues included a lymph node with angioimmunoblastic lymphadenopathy and a hyperplastic tonsil. Weak positive staining for ELAM-1 was found in 1 patient in vessels, both with and without leukostasis. Expression of VCAM-1 and ICAM-1 in all patients tested was similar to that in the negative controls. The results of this study suggest that activation of endothelium, with increased expression of the endothelial adhesion molecules under study, is not a prerequisite for the development of pulmonary leukostasis in leukemia. PMID- 8237272 TI - Chronic idiopathic thrombocytopenic purpura complicated by chronic disseminated intravascular coagulation associated with abdominal aortic aneurysm. AB - A patient with chronic idiopathic thrombocytopenic purpura (ITP), chronic disseminated intravascular coagulation (DIC) and abdominal aortic aneurysm is described. He had severe thrombocytopenia and his platelet life span was shortened (29 h). Indium 111-labeled platelet scintigraphy demonstrated an increased accumulation of radioactivity over the aneurysm and the spleen. Following preoperative heparin therapy for DIC, the platelet count remained low (10 x 10(9)/l), while the DIC improved. Because of the elevated platelet associated IgG, we considered that the patient had complicated chronic ITP. After intravenous immunoglobulin therapy, his platelet count increased. The aneurysm was successfully replaced with an artificial Y-graft vessel and a splenectomy was performed at the same time. The platelet count was maintained at about 60 x 10(9)/l after the operation and the patient is quite well now. PMID- 8237273 TI - Aplastic anemia and idiopathic thrombocytopenic purpura with antibody to platelet glycoprotein IIb/IIIa following resection of malignant thymoma. AB - A case of a 64-year-old Japanese woman who developed aplastic anemia and idiopathic thrombocytopenic purpura with antibody to platelet glycoprotein IIb/IIIa 4 years following a resection of malignant thymoma is reported. Bone marrow was hypocellular and ferrokinetics revealed the prolongation of the half time of plasma iron disappearance and a decrease in red cell utilization, findings compatible with a diagnosis of aplastic anemia. The life span of platelets was markedly decreased to 3.07 h, and the test for antiplatelet glycoprotein IIb/IIIa antibody was positive. The patient's serum had a suppressive effect on the formation of colonies of burst-forming unit-erythroid and colony-forming unit-granulocyte using normal bone marrow cells. These results suggest that the aplastic anemia in this patient may have been induced by some suppressive activity in the serum. Splenectomy followed by an administration of cyclosporine effectively restored the peripheral blood count. PMID- 8237274 TI - Spontaneous regression in B cell, diffuse large cell type non-Hodgkin's lymphoma. AB - A rare case of non-Hodgkin's lymphoma (NHL) that showed transient spontaneous regression (SR) is described. After 6 months of remission, recurrence was noted in the lymph nodes, pleura and the spleen. Although transient improvement was observed following combination chemotherapy, the pleural effusion became refractory to chemotherapy and the patient died 9 months after the relapse. PMID- 8237275 TI - Low-grade lymphoma following intensive treatment of large-cell lymphoma. AB - Progression of a low-grade non-Hodgkin's lymphoma into a more aggressive histologic pattern is a well-described phenomenon. The converse phenomenon, often called downgrading, is much less frequent. We report 2 patients in whom relapse with an indolent lymphoma was noted after chemotherapy for higher-grade disease. Other previously reported cases are reviewed. The implications for appropriate management and pathogenesis are discussed. PMID- 8237276 TI - Analysis of BCR/ABL abnormalities in mRNA from 20-year-old paraffin-embedded tissue for BCR/ABL rearrangement by polymerase chain reaction. AB - We studied whether it is possible to obtain sufficient mRNA from old paraffin embedded samples of spleen and bone marrow for reverse transcription and polymerase chain reaction analysis. Spleen tissue from 6 subjects with chronic myelogenous leukemia was studied for rearrangement of BCR and ABL genes. Analysis was successful in 4 cases. These data indicate the feasibility of retrospective analysis of tissue samples of special interest by molecular techniques. PMID- 8237277 TI - Massive tissue necrosis can be induced by heparin. PMID- 8237278 TI - Retrospective study on the influence of human parvovirus B19 infection among children with malignant diseases. AB - Human parvovirus B19 (B19) is a known cause of erythema infectiosum (fifth disease) and aplastic crisis in patients with hemolytic anemias. When patients with malignant diseases are infected by B19 during chemotherapy, erythroid suppression of bone marrow sometimes occurs. We performed a retrospective investigation of B19 infection among 95 children with malignant diseases in our hospital during the past 14 years. By the method of dot blot hybridization, 9 of 95 patients were found to be positive for B19 DNA during chemotherapy. All 9 patients had reticulocytopenia at the time B19 DNA was detected in their serum samples. Neutropenia and thrombocytopenia were not found. Seven of them had only transient reticulocytopenia. Serum samples from 2 other patients were positive for B19 DNA for a longer time. They suffered from persistent anemia for about 2 and 13 month, respectively. The years when B19 DNA was detected from the 9 patients corresponded to the prevalence of erythema infectiosum in Japan. PMID- 8237279 TI - [Bacterial endocarditis: microbiological diagnosis]. AB - The microbiological laboratory can assist in diagnosis and therapy of bacterial endocarditis with the following parameters: cultivation and identification of the causative organism, sensitivity testing including assessment of the minimal bactericidal concentration (MBC) and testing of antibiotic combinations with checkerboard titrations and time-kill curves. Serum bactericidal titers, although controversial with respect to their utility in prognosing the clinical outcome, are regarded useful by some authorities but not by others. Antibiotic assays, mainly for aminoglycosides and vancomycin, serve to monitor antibiotic serum levels. PMID- 8237280 TI - [Continuous measurement of mixed venous oxygen saturation]. AB - The task of the cardiorespiratory system is to deliver enough oxygen to meet the metabolic requirements of the body. Of all metabolic substrates, oxygen has the highest percentage of extraction and oxygen reserves are exhausted within a few minutes. Arterial oxygen content and cardiac output are the determinants of oxygen delivery (DO2). O2-delivery in intensive care patients can be compromised by many factors, especially diminished O2-uptake in the lung, reduced hemoglobin content, and/or altered cardiac performance. To calculate DO2, determination of cardiac output (CO), O2-tension, O2-saturation, and hemoglobin is necessary. O2 consumption (VO2), the best mirror of the actual global metabolic activities of the tissues, can be measured invasively using arterial-venous O2-content difference (avDO2) and cardiac output, which requires pulmonary artery catheterization. The body has two basic mechanisms to meet increased VO2 or to compensate for a decrease in DO2: an increase in CO and a rise in O2-extraction ratio. The avDO2 and O2-extraction ratio are known to be very good indicators for the adaptation of the cardiorespiratory system to the actual metabolic requirements of the body. The oxygen supply-to-demand ratio (DO2/VO2) yields information on the extent to which these compensatory mechanisms either function or fail. It was demonstrated that changes of the avDO2 and changes of the DO2/VO2 are always paralleled unidirectionally by changes of mixed venous O2-saturation (SvO2). Therefore these parameters can easily be replaced by SvO2 for clinical purposes. Continuous real time in-vivo control of SvO2 has now become possible using commercially available devices with fiberoptic catheters. Pathophysiological changes in the O2-supply-to-demand-ratio as well as effects of ongoing therapy can be determined at the bedside. PMID- 8237281 TI - [Valve saving correction of aneurysms of the ascending aorta with aortic insufficiency or annular dilatation]. AB - The current technique to correct aneurysms of the ascending aorta with incompetence of the aortic valve or dilatation of the aortic root is the implantation of a valved conduit. In many cases however structurally normal aortic leaflets are resected by this technique and patients are at the risk of complications due to mechanical heart valve prostheses and the necessary anticoagulation respectively. By resuspension of the aortic valve into the aortic prostheses similar to the technique used at implantation of homograft valves, competence of the valve can be restored and the dilatation of the aortic annulus can be prevented or even reduced. Our article describes the successful, valve sparing correction of an acute aortic dissection type A with dilatation of the aortic annulus in a 43 year old patient and in another patient with ascending aortic aneurysm with aortic insufficiency. PMID- 8237282 TI - [Oxidized lipoproteins and atherogenesis]. AB - The assumption that the oxidation of lipoproteins plays an eminent role in the development of atherosclerosis is of increasing significance. Our examinations concerned the kinetics of lipid peroxidation of the poly-unsaturated fatty acids in lipoproteins and the existing protection mechanisms by antioxidants, such as vitamin E and carotenoids. Clinical studies proving a correlation of plasma levels and antioxidants, such as vitamin C and E and carotenoids in correlation with cardiovascular disease are cited and discussed. Certain aldehydes originating during lipid peroxidation in LDL modify the protein part of LDL. The consequence of the oxidative modification of LDL is a decreased recognition of LDL by the LDL-receptor and moreover the uptake by macrophages hereby changing to foam cells. Epitopes as developed in oxidatively modified LDL were also shown immunohistochemically in atherosclerotic plaques. Finally a hypothetic scheme is presented how LDL is oxidized in the vessel wall and how it initiates processes resulting in atherosclerosis. PMID- 8237283 TI - [The role of current diagnostic measures in hypertension]. AB - The problems in diagnostic steps in the hypertensive patient can be grouped in 4 categories: the etiological evaluation concentrates recently especially on renovascular hypertension since. This disorder is getting more common with the ageing population and since diagnosis and treatment have been simplified; the detection of additional risk factors is important to guide individual drug selection; the detection of organ manifestations (e.g. LVH) may help in the therapeutic decision especially in borderline hypertensives; ambulatory blood pressure monitoring is becoming increasingly used despite the lack of standardized and generally accepted means to evaluate the recorded data. PMID- 8237284 TI - [Significance of treatment of early forms and prevention of heart failure. Value of ACE inhibitors in therapy after myocardial infarct]. AB - Myocardial infarction has become the most important condition causing congestive heart failure. Several treatment regimens compete with each other in the early and later post myocardial infarction phase focussing on different pathophysiologic mechanisms. In this review emphasis is put on absolute risks and absolute risk reductions in patients with left ventricular dysfunction after myocardial infarction. In the first year mortality is still as high as 10% and the major risks are sudden unexpected death, reinfarction, and congestive heart failure. In the following years sudden death and development of congestive heart failure become less prominent, while a considerable risk of reinfarction persists. Thrombolysis and beta-blockade are the most important drugs in the acute infarction phase leading to a marked reduction in one year mortality. In the first year after infarction beta-blockade has an established place in the prevention of sudden death; revascularization and aspirin are established in reinfarction prophylaxis. However, since most drugs lower 1 year mortality by 0.5 to 2% only, drugs like ACE inhibitors that influence more than one risk, i.e. sudden death and reinfarction as well as deterioration of pump dysfunction might play a more important role in the therapeutic strategy after myocardial infarction. A supplementation by one or two drugs aiming at the individually estimated leading risks might be the best choice in patients with left ventricular dysfunction today. PMID- 8237285 TI - [Coronary artery dilatation after infarct]. AB - International studies (TAMI, TIMI II A, TIMI II B, European Cooperative Study, DANAMI, GUSTO, LATE, SWIFT, SAVE) confirmed the concept of coronary balloon angioplasty (PTCA) after acute myocardial infarction to be reduced to strict indications. These strict indications are on the one hand side based on anatomically suited coronary lesions, on the other hand side dependent on ischemia and/or angina. Direct (immediate) PTCA, rescue PTCA in evolving infarctions after failing lytic therapy are recommended, if operators and a well trained team are available on a 24 hours basis. Routine PTCA after myocardial infarction is not save enough any time after myocardial infarction with or without lysis, therefore "watchful waiting" (Braunwald) still is recommended. But ischemia also depends on how much you look for it! The state of total occluded vessels, significant stenoses without ischemia, the elderly, non Q-wave infarctions, reduced left ventricular function are a field of discussion and patients with these findings should rather undergo angioplasty and PTCA, as long as we have no other detailed results from ongoing studies. PMID- 8237286 TI - AARN initiatives for health care reform: what they are and how to get involved. PMID- 8237287 TI - Task force: direct access to services provided by registered nurses. PMID- 8237288 TI - NBSS Part 1: Does mammography screening provide a significant benefit to women aged 40-49? PMID- 8237289 TI - Dr. Verna Huffman-Splance, Dr. Richard Splane: an interview regarding their research study. Senior nurses in government: chief nursing officer positions in national ministries of health. Interview by Eve Henderson. PMID- 8237290 TI - Gerontological nursing education for Alberta nurses. PMID- 8237291 TI - Independence in nursing: reclaiming the past. PMID- 8237292 TI - Car '93--where are you? PMID- 8237293 TI - You have been laid off: what's next! PMID- 8237294 TI - Dinosaurs at Foothills Hospital: nursing research influences hospital wound care procedures. PMID- 8237295 TI - Bedside evaluation of intraocular pressure in critically ill patients, ventilated at different levels of positive end-expiratory pressure. AB - Animal experiments suggest that the application of positive end-expiratory pressure (PEEP) levels > or = 10 cm H2O increase intraocular pressure (IOP), probably through ophthalmic and episcleral venous outflow obstruction secondary to PEEP-induced increase in central venous pressure (CVP). To evaluate whether a similar response occurs in humans, we studied the effects of varying levels of PEEP on IOP in 11 critically ill sedated and mechanically ventilated patients, aged 35 to 88 yrs (mean: 68 yrs), without evidence of ocular disease. Measured variables included PaCO2, PaO2, mean arterial pressure (MAP), CVP and IOP, and were recorded at zero end-expiratory pressure and at 5, 10 and 15 cm H2O PEEP, applied in random order. IOP was measured by the same investigator at the bedside, using a portable and battery-operated tonometer (Tono-Pen). As expected, PaO2 increased significantly from baseline at all PEEP levels. At the three levels of PEEP, no significant change of PaCO2 from its corresponding baseline values was observed. At 5 cm H2O PEEP neither CVP nor IOP raised significantly from baseline. IOP increased significantly (p < 0.01) from 12 +/- 4 to 14 +/- 4 mmHg at 10 cm H2O PEEP and from 13 +/- 4 to 16 +/- 5 mmHg at 15 cm H2O PEEP. CVP also increased significantly (p < 0.01) and in parallel with IOP at 10 and 15 cm H2O PEEP. Since IOP tends to rise significantly when PEEP levels exceed 10 cm H2O, PEEP-ventilated patients in whom such increase is undesirable might benefit from regular bedside IOP monitoring. PMID- 8237296 TI - A rare complication of pulmonary artery catheterization during cardiac surgery: entrapment of pulmonary artery catheter around an intracardiac structure. PMID- 8237297 TI - Monoamine oxidase inhibitors and anesthesia: an updated literature review. AB - Safe anesthesia for patients taking MAOIs has been possible since the 1980s using judicious choice of agents. The introduction of a new class of MAOIs (RIMA) has increased the margin of safety for these anesthetics. These drugs allow selective and rapidly reversible inhibition of type A-MAO. The pharmacologic effects of this new class of drug as well as their potential interactions with agents used in anesthesia are reviewed. PMID- 8237298 TI - The vicarious liability of a surgeon for a negligent anesthesiologist. AB - In June 1992, the issue of whether a surgeon could be held liable for the negligence of an anesthesiologist was examined by the tribunal of Leuven (1). Liability was asserted not only against the negligent anesthesiologist, but also against the surgeon. The judgement is most interesting and raises many controversial points of law. Although, in most jurisdictions today, it is recognized that the surgeon is no longer the captain of the ship, but shares the responsibility with other colleagues such as the anesthesiologist, the Belgian judge, free as the trier of fact, can exceptionally hold a member of a surgical team liable for the negligence of his colleague. A critical analysis of this and two other recent cases in Belgium forms the basis of this paper, which deals with the liability of the surgeon for the anesthesiologist's negligence. This article will centre around four issues: First, the nature of the contractual relationship between anesthesiologist and patient. Second, the liability (criminal and civil) of a team member for his own fault if he observes a negligent colleague and does not take the appropriate measures. Third, the liability in Belgian tort law of the surgeon for the negligent anesthesiologist. Fourth, the liability in Belgian contract law of the surgeon for the anesthesiologist. PMID- 8237299 TI - [Buddhist mummies in Japan]. AB - The mummy of priest Kochi (preserved at Saishoji Temple, Teradomari, Niigata Pref.) has become famous, since it appeared in the book "Snow Country Tales" written by Bokushi Suzuki in 1841 (Fig. 1). In a country of high humidity, such as Japan, the belief that mummification could not, and did not, exist would not be altogether unfounded, but rather more a matter of common sense. There are two dozen Buddhist mummies in this country. It was not known until 1961 that a reliable source of artificial mummification has existed in Japan. The Japanese Buddhist mummies, apart from those of the Fujiwara family, a powerful clan of northeast Japan in the 12th century, dated mostly from the 17th to the 19th century as given in Table 1. Three principal types of mummification described by Vreeland, Jr. and Cockburn (1980) could be identified in the Japanese Buddhist mummies: type I, natural mummification; type II, intentional natural; and type III, artificial. Matsumoto (1990) classified the mummies into four groups, based on their ideological backgrounds: group A, mummies of the priests having faith in the Amitabha (the Supreme Buddha presiding over the Pure Land in the West); group B, sokushin-butsu mummies of the priests belonging to the Shingon sect of Japanese Buddhism founded by Kukai (Kobo-daishi); group C, nyujo mummies of the priests having faith in the Maitreya (the Buddha presiding over the Pure Land in the North, or the Buddha of the future); and group D, other mummies. These mummies of groups A, B, C and D are respectively listed in Table 2. Previous papers have shown that the mummies of the groups A, C and D belonged to the mummification of type I (natural mummification) or type II (intentional natural), whereas those of only the group B were of type III (artificial). The mummies of groups A to D were given as follows. a) Mummies of group A. The four mummies of the Fujiwara family in the Amitabha faith (preserved at Chusonji Temple, Hiraizumi, Iwate Pref.), which were laid in daisies for Buddhist images at golden Konjikido, a model of the Western Pure Land of Amitabha, did not bear any evidence of Ainu character in their physical traits. Of these, the mummy of Yasuhira Fujiwara, of which the head and neck were severely injured by swords, showed that it had been decapitated. This decapitated head had suffered mutilations: i.e. a long iron nail had been driven into the forehead, and the both auricles and the nose had been cut off.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8237300 TI - Current studies on tooth enamel development in lower vertebrates. AB - The tooth enamel is a unique tissue in the human body on many characteristic features, in both morphological and developmental aspects. This tissue is widely distributed in the vertebrate world. This hard tissue is divided into two groups, mainly from the developmental feature. One is the enamel in the upper classes of the adult amphibian, called true or ectodermal enamel. The other mainly distributes in fish (both of osteichytyes and chondroichtyes), called enameloid or mesodermal enamel. Enameloid seen in fish has many characteristics compared with enamel, although enameloid after eruption is analog tissue to enamel. Many studies on the enameloid with a long history show its character. (1) The enameloid matrix is mainly composed of collagen fibrils to make thick bundles, most of which are interwoven in a complicated manner. (2) Mineralization of enameloid is initiated by the matrix vesicles similar to bone and dentine. (3) Growth of crystals does not stop the retention of their thin needle shape as with bone and dentine during mineralization, but finally begins to show a large hexagonal shape, as seen in the mature enamel of mammals. (4) Enameloid contains much fluoride and the fluoride exists as a fluoroapatite in enameloid. Many of the recent studies on this subject applied various histochemical methods, especially immunohistochemical studies, which show some evidence that enameloid and enamel might not be continuous in enamel evolution. It is now necessary to reconsider the enameloid from the phylogenetic aspect. PMID- 8237301 TI - [Arterial embalming method of the cadaver and its application to research]. AB - Ever since 1974, the cadaver has been embalmed by the arterial embalming method, using pre-embalming fluid with blood clot disperser and cell conditioner for the removal of blood clots and drainage of blood, at the Department of Anatomy of the Kawasaki Medical School. According to this method, the cadavers are always very well fixed so that they can be used for not only anatomical dissection but also research for the vascular system by vasography, kinematics of the joint and other histologic examinations. In this report we have described our embalming procedure concretely and its application to research. PMID- 8237302 TI - Anomaly of bilateral persistent sciatic arteries. AB - A pair of persistent sciatic arteries was encountered in the body of a 91-year old Japanese woman in the course of an ordinary dissection at Nara Medical University. The artery of each side followed nearly the same course in the posterior aspect of the thigh; after leaving the pelvis through the infrapiriform foramen, the persistent sciatic artery was observed descending between the tibial and common peroneal nerves, accompanied by the sciatic vein. In the popliteal fossa, the persistent sciatic artery ended by joining the popliteal artery. The sciatic nerves on both sides were found to have divided, while in the pelvis, into the tibial and the common peroneal nerves; the former passed through the infrapiriform foramen, while the latter perforated the piriformis muscle to reach the posterior aspect of the upper thigh. PMID- 8237303 TI - Clinical experience with glass ionomer for proximal fillings. AB - The aim of this study was to evaluate, with the aid of a questionnaire handed out to a selected group of dentists, the use of glass ionomer cement (GIC) in different types of proximal preparations. The aim was to evaluate the experience of complications associated with the use of GIC. Very few had often observed secondary caries or gingival inflammation in association with GIC fillings, compared with about 70% of the dentists in association with posterior composites. Tunnel fillings had been made by 60% of the dentists, simple proximal fillings in primary molars by 80%, and sandwich restorations by 80%. Few dentists with at least 2 year's experience with tunnel fillings had observed biologic complications, but ridge fractures had often been observed by 10%. Among the dentists with at least 2 years' experience with proximal fillings in primary molars, 40% mentioned more complications with these than with amalgams. Biologic complications were also not a great problem with GIC/composite sandwich restorations, but wear or dissolution of the proximal GIC surface was often seen by 17% of the dentists. PMID- 8237304 TI - The fit of gold inlays and three ceramic inlay systems. A clinical and in vitro study. AB - Four inlay systems--gold, Cerec, Mirage, and Empress inlays--were evaluated for their adaptation to stone die and clinically to the tooth by means of a replica technique. Twenty inlays of each system were placed on premolars and molars in the lower jaw. A microscope was used to measure the adaptation at the approximal margin, at the inner axial wall, and at the occlusal cavosurface area. An overall better fit was observed for the gold inlays than for the ceramic inlays. When the different measuring locations were compared, a better fit was found for the occlusal area. The greatest discrepancies were recorded for the Cerec inlays, whereas the Mirage and Empress inlays were comparable. PMID- 8237305 TI - The ultrastructure of copper amalgam-covered dentin from human deciduous teeth. AB - The structure of dentin subjacent to Cu-amalgam restorations was studied. The restorations had been inserted 7-8 years before extraction of the teeth. No lining had been applied. The dentin subjacent to the restorations had a greenish grayish discoloration, and in the pulpal end irregular secondary dentin had formed. Energy-dispersive X-ray microanalysis of areas showing discoloration confirmed the presence of Cu. Semithin and ultrathin undemineralized sections were studied. Vital pulp tissue and irregular secondary dentin were observed. Close to the predentin and halfway into the dentin most of the dentinal tubules were occluded. In the control material only an occasional occluded tubule was seen. Close to the Cu-amalgam restoration some tubules were occluded or partly occluded, and some were open. The odontoblasts had responded to the irritating agent by obturation of the tubules, thus reducing the permeability of the dentin. Furthermore, irregular secondary dentin had formed in another attempt to wall off the irritating agent. PMID- 8237306 TI - Polishing systems for dental ceramics. AB - The increasing use of ceramic crowns and inlays in posterior teeth has highlighted the clinical difficulties involved in achieving a good surface finish after corrective grinding. In the present work several polishing systems and techniques were compared, initially using only one dental ceramic, Vita Mark I. The two systems that produced the best results were then further used for the polishing of several different dental ceramics, mainly newer types. The initial surface roughness and the surface condition attained after various polishing stages were evaluated by using a surface roughness analyzer. In the initial test series the SofLex system and the Shofu Porcelain Laminate Polishing Kit produced the best results; they were therefore chosen for the second part of the investigation. In this the two polishing systems concerned produced a satisfactory surface finish in a clinically acceptable time for all the ceramics tested. The use of a diamond-containing polishing past did not improve the surface smoothness obtained with the SofLex system but gave a more varied result for the Shofu system. PMID- 8237307 TI - Adherence of resin-based luting agents assessed by the energy of fracture. AB - The present study assessed the adherence of nine commercial resin cements by means of the wedge test. The beams of the test were made of a Ni-Cr-Be alloy, and the adhering surfaces were sandblasted with 250 microns Al2O3. The energy of fracture of the investigated cements varied from less than 10 J/m2 to 121 J/m2. The fracture energies were not influenced by 1) the thickness of the joint, 2) the width of the beams, 3) the use of an intermediary noncomposite resin, or 4) storage in water beyond 24 h. In spite of a rather high variability within groups, it is concluded that the wedge test may give relevant data on the performance of adhesive joints. PMID- 8237308 TI - Xylitol-induced changes of enamel microhardness paralleled by microradiographic observations. AB - The study, aimed to analyze the feasibility of a prospective field study, was carried out in Polynesian children with rampant untreated caries. Slabs of bovine enamel were inserted for 8-12 days in cavities and subsequently replaced by permanent fillings. Before use, the surface of the slab was polished, and one half predemineralized and tested for microhardness. The follow-up in 30 subjects involved 54 slabs, 30 from negative controls with no added sweets and 24 from subjects receiving 20 g/day of xylitol in candy. The microhardness of the slabs was reassessed, and the difference between measurements calculated and tested for significance. The differences between the groups were highly significant, the predemineralized halves showing pronounced rehardening at exposure to xylitol. Parallel microradiographic observations conformed with the above findings. The results indicate that the use of a noncariogenic sweetener might be of value in high caries risk subjects. PMID- 8237309 TI - Quality of fixed prosthodontics after 15 years. AB - Interviews and/or clinical examinations by means of the California Dental Association quality evaluation system were carried out in a group of persons who had received extensive restorative treatments with fixed partial dentures 15 years before this study. The studied group consisted of 77 persons who agreed to participate from an original group of 150 persons selected at random from the Swedish Dental Insurance System records. Of the original group 20 had died, 17 were not traceable or not able to participate for medical reasons, and 36 declined to participate. Thirty-two per cent of the recorded reconstructions had been lost, and 8% partially lost during the 15-year period. Thirty-five per cent of the reconstructions were rated as Satisfactory, whereas the remaining ones had mixed clinical quality ratings. Failures and Not Acceptable quality ratings were found to be caused mainly by fractures, loss of retention, and/or dental caries. PMID- 8237310 TI - 11-year assessment of Class-III resin restorations completed with two restorative procedures. AB - This randomized clinical trial was initiated to study the effect of two restorative procedures on the clinical quality and longevity of class-III resin restorations. The material consisted of 52 pairs of class-III restorations in the microfilled resin Silar completed with two modifications of the conventional acid etch restorative procedure: A) beveling of the margins of the cavity preparations, and B) treatment of the etched butt-joint cavities before filling with the dentin-adhesive NPG-GMA/ethanol together with re-etching and surface coating of the finished restorations with low viscous resin. The restorations were examined at base line and after 2, 4, 6, and 11 years. The cumulative 11 year survival rate for both types of restorations was 84%. Marginal discolorations, marginal discrepancies, and secondary caries were most often recorded corresponding to the beveled type-A restorations, whereas the esthetic quality of the two types of restorations was almost identical. Surface discolorations were most often recorded among smokers, and significant correlations were found between the patients' consumption of alcoholic beverages and corpus discoloration, surface discoloration, and surface wear of restorations. PMID- 8237311 TI - In vitro quality testing of dentin adhesives. AB - A tensile and a shear bond strength test are compared, using two well-known and two more recently marketed dentin adhesive systems: Gluma (Bayer), Scotchbond 2 (3M), Scotchbond Multi-Purpose (3M), and Syntac (Vivadent). The adhesives were used to bond composite resins to the buccal surface of human third molars. Specimens stored for 24 h before testing did not show significant differences between bond strength values obtained by either the tensile or the shear test method for any of the materials. The bond strength of thermocycled specimens did not differ for Gluma, was significantly lower for Scotchbond 2, and significantly higher for Scotchbond Multi-Purpose and Syntac. SEM observations showed increasing amounts of cohesive failure in resin with increasing bond strength values. Cohesive fractures in dentin were also observed. It is concluded that either the tensile or the shear test method can be used for quality testing of dentin adhesives. Thermocycling may provide interesting information about the quality of a bond. PMID- 8237312 TI - Prognosis of total hip replacement in Sweden. Follow-up of 92,675 operations performed 1978-1990. AB - A prospective, national multi-center study of all reoperations after total hip replacement (THR) was started by the Swedish Orthopedic Association in 1979. The material comprises all THR performed in Sweden, presently more than 10,000 yearly or 130 THR per 100,000 inhabitants; uncemented implants have been used in less than 2 percent. The main reasons for revision have been aseptic loosening 79 percent, infection 10 percent, technical error 6 percent, and dislocation 2 percent. The cumulative rate of revision for deep infection has dropped from 0.9 percent to < 0.5 percent for implants inserted 1979 and 1983, respectively. With the Charnley prosthesis as the gold, standard the performance of other prostheses was analyzed. Improved cementation techniques and anti-infection measures have continuously reduced the revision risk. The register demonstrates that the average orthopedic surgeon cannot match the results achieved by experts. However, the vast majority of THR, worldwide, are not performed by experts. Quality assurance in this sector of orthopedics demands a continuous analysis of the outcome of these operations. PMID- 8237313 TI - Wear of polyethylene acetabular cups against alumina femoral heads. 5 prostheses compared in a hip simulator for 35 million walking cycles. AB - 5 ultra-high molecular weight polyethylene acetabular cups articulated against alumina femoral heads for 35 million walking cycles in a hip joint simulator designed for wear tests of total hip prostheses. The specimens were from Protek, Biomet, Link, Howmedica and Thackray. In the Howmedica specimen, the wear was zero, and in Link and Thackray practically negligible. In Biomet, however, the wear was noteworthy and in Protek disastrous after 20 million cycles because in these 2 prostheses the head was attached to a titanium-alloy stem by taper-fit: titanium-alloy particles that were removed from the taper because of micromotion between the head and stem were entrapped between the head and cup, adhering to the head and making it rough, which led to severe abrasive wear of the Protek cup. It was worn through at 26 million cycles, the total wear being 3,170 mg. In Biomet, Link and Thackray, the total wear was 124, 5.3 and 17.6 mg, respectively. Polyethylene wear particles may lead to adverse tissue reactions and eventual loosening of the implant. The results indicate that by the use of alumina heads, polyethylene wear can be eliminated, but this advantage may be lost if the head is attached to a titanium-alloy taper. PMID- 8237314 TI - Association of overweight, trauma and workload with coxarthrosis. A health survey of 7,217 persons. AB - In a population sample health survey, body mass, previous trauma and physical stress were studied for associations with coxarthrosis. A sample of 8,000 persons representative of the Finnish population aged 30 or over was invited for examination, and 90 percent participated. On the basis of a standardized clinical examination, a physician diagnosed coxarthrosis in 6 percent of the women and 4 percent of the men. The prevalence rose with age. In persons with a past traumatic lower-limb injury, the odds ratio of unilateral coxarthrosis was 2.1 and of bilateral coxarthrosis 1.5, as adjusted for sex, age and other determinants using logistic regression. The sum index reflecting self-reported features of physical stress in present or previous occupations was directly proportional to the prevalence of coxarthrosis. Body mass index (kg/m2) was closely associated with bilateral coxarthrosis; the adjusted odds ratio (95 percent confidence intervals) for indices > 35, compared to those < 25, was 2.8 (1.4-5.7). In terms of the population attributable fraction, prior trauma, physical stress and body mass were estimated to explain 59 percent of the prevalence of coxarthrosis. The potential for primary prevention may be great, but longitudinal population studies are necessary to elucidate causal significance of the risk factors. PMID- 8237315 TI - A new surgical approach in 8 cases of polygonal triple pelvic osteotomy. AB - We have undertaken 12 polygonal osteotomies, using an improved approach in 8. The operation was successful in correction of acetabular dysplasia after ossification of the triradiate cartilage. It was also useful in early arthrosis with slight deformity of the femoral head. PMID- 8237316 TI - Increased blood loss after preoperative NSAID. Retrospective study of 186 hip arthroplasties. AB - We have evaluated bleeding during and after hip replacement in 186 patients in relation to preoperative intake of nonsteroidal anti-inflammatory drugs (NSAID) combined with low molecular weight heparin. NSAID was associated with increased preoperative bleeding and blood transfusion requirements. PMID- 8237317 TI - Plasma and bone concentrations of cefuroxime and flucloxacillin. Oral versus parenteral administration in 20 arthroplasties. AB - Our objective was to determine and to compare the range of bone levels of cefuroxime and flucloxacillin achieved after oral and intravenous administration in 20 arthroplasty patients, allocated to 4 groups: 1 x 500 mg or 7 x 500 mg oral cefuroxime was followed by 2000 mg Flucloxacillin i.v.; 1 x 500 mg and 7 x 500 mg oral flucloxacillin was followed by 1500 mg cefuroxime i.v. Bone samples of hip and knee were obtained. Oral administration did not result in a measurable bone concentration of any of the antibiotics. Intravenous administration resulted in measurable bone concentrations of both cefuroxime and flucloxacillin, with large inter-individual variations. The bone concentrations of intravenous cefuroxime were higher than those of flucloxacillin, despite the lower dose. Oral pretreatment had no effect on the bone concentrations after intravenous administration. No accumulation of the drugs in bone was observed. PMID- 8237318 TI - Periarticular bone mineral content in rheumatoid arthritis and arthrosis of the hip. Dual photon X-ray absorptiometry in 53 cases. AB - The bone mineral content (BMC) was determined by dual photon X-ray absorptiometry prior to hip replacement in 22 patients with rheumatoid arthritis and 31 with arthrosis. Compared with the arthrosis patients, the rheumatoids had approximately 20 percent lower bone mineral content in the proximal femur, the lumbar spine, and in the innominate bone superior to the acetabulum. PMID- 8237319 TI - Blood perfusion uneven in femoral head osteonecrosis. Doppler flowmetry and intraosseous pressure in 12 cases. AB - We measured the microvascular regional perfusion in 12 hips with non-traumatic osteonecrosis of the femoral head, using laser Doppler flowmetry (LDF). Simultaneously, the intraosseous pressure (IOP) was measured in the femoral head. For comparison, the same 2 parameters were measured in 6 normal femoral heads during surgery of the contralateral hip. In osteonecrosis, the regional blood cell flux in the intertrochanteric area was 165 mV, at the rim of the lesion 430 mV, and in the necrotic lesion 35 mV. In the same areas, the IOP was 38 mmHg, 61 mmHg and 55 mmHg. In the normal hips, the LDF signal was 221 mV and 224 mV, and the IOP was 21 mmHg and 19 mmHg intertrochanterically and in the femoral head, respectively. We conclude that the microvascular blood perfusion is uneven in an osteonecrotic head. PMID- 8237320 TI - Magnetic resonance imaging not indicated in healed Perthes' disease. Comparison with radiography in 10 cases. AB - We examined 10 patients with unilateral healed Perthes' disease with plain radiographs and magnetic resonance imaging (MRI). The mean time between the onset of symptoms and follow-up was 8 (4-10) years. The MRI scans did not contribute more information than the plain radiographs. PMID- 8237321 TI - Urban vs rural increase in hip fracture incidence. Age and sex of 901 cases 1980 89 in Olmsted County, U.S.A. AB - To determine if hip fracture incidence rates are greater in urban than in rural areas of the United States, as they appear to be in Scandinavia, we undertook a study in Olmsted County, Minnesota. During the decade 1980-89, the age- and sex adjusted incidence of proximal femur fractures among urban residents of the central city of Rochester was 36 percent greater than among residents of the rural remainder of Olmsted County. The difference was almost entirely accounted for by an excess of fractures due to moderate trauma in elderly urban women. These first detailed results from the United States confirm earlier observations from Scandinavia that hip fracture rates are lower in rural areas. PMID- 8237322 TI - Early complications after anterior dislocation of the shoulder in patients over 40 years. An ultrasonographic and electromyographic study. AB - The rate of complications after anterior dislocation of the shoulder was evaluated in 65 patients aged over 40 years. 36 of 55 cases had electromyographically verified axillary nerve or brachial plexus injury. Rotator cuff lesion was seen in 24 of the 63 sonographically examined cases. At follow-up in a telephone interview on average 3 years after the injury, 27 of the 57 cases had complaints from their shoulder. The incidence of initial nerve and/or cuff lesions was higher in those with persisting symptoms at follow-up. PMID- 8237323 TI - Transforming growth factor-beta enhances fracture healing in rabbit tibiae. AB - The ability of exogenous Transforming Growth Factor-beta (TGF-beta) to stimulate bone formation in fracture healing was investigated. TGF-beta was continuously applied in doses of 1 and 10 micrograms/day for 6 weeks to 2 groups of adult rabbits with unilateral plated mid-tibial osteotomies. A group receiving solvent without TGF-beta served as control. Fracture healing was evaluated by mechanical tests, bone morphometry and bone densitometry. Increased maximal bending strength and callus formation were demonstrated in the groups receiving TGF-beta. TGF-beta had no effect on bending-stiffness, bone mineral content, cortical thickness or haversian canal diameter. We conclude that local application of exogenous TGF beta may enhance fracture healing in rabbits. PMID- 8237324 TI - Basic fibroblast growth factor and bone induction in rats. AB - Bone induction is initiated by bone morphogenetic proteins, but local growth factors present in demineralized bone matrix (DBM) may further regulate the process. We have previously shown that local application of recombinant human basic fibroblast growth factor (bFGF) in a carboxymethyl cellulose gel to DBM implants increases the bone yield, as measured by calcium content. In the present study, similar experiments were evaluated with histomorphometry. The chondrocyte number at 2 weeks was increased by the application of 15 ng bFGF. This increase was due to an increased number of chondrocyte clusters, i.e., cartilage formation was initiated in more places within the implant. The size of the individual chondrocyte clusters was the same as in the controls. Thus, the bFGF had probably stimulated cellular events preceding chondrocyte proliferation. At 3 weeks, the chondrocytes were fewer than in controls, and instead there was more bone. Thus, cartilage formation was increased by bFGF, and its replacement by bone came earlier. However, 1900 ng of bFGF had a profound inhibitory effect on both cartilage and bone formation. PMID- 8237325 TI - Enhancement of bone formation in rabbits by recombinant human growth hormone. AB - We studied the effect of human recombinant growth hormone on diaphyseal bone in 40 adult rabbits. The diaphyseal periosteum of one femur in each animal was mechanically stimulated by a nylon cerclage band. The bands induced an increase in bone formation, bone mineral content, and maximum torque capacity of the diaphyseal bone at 1 and 2 months. Growth hormone enhanced the anabolic effect of the cerclage bands on bone metabolism, evidenced by a further increase in torsional strength of the femurs. PMID- 8237326 TI - Laxity after cruciate ligament injury in 94 knees. The KT-1000 arthrometer versus roentgen stereophotogrammetry. AB - We examined 94 knees with chronic anterior cruciate ligament injuries, 55 of which had been operated with ligament reconstruction, using the KT-1000 arthrometer (89 N anterior force) and roentgen stereophotogrammetry (RSA, 150 N anterior, 80 N posterior force). In intact knees the tibial displacement did not differ between the methods. In injured knees, operated or not, the KT-1000 recorded smaller AP translations and side-differences than RSA. Thus, the stabilizing effect of reconstructive surgery may be overrated, if evaluated with the standard KT-1000 technique. PMID- 8237327 TI - Effect of femoral condyle configuration on disability after an anterior cruciate ligament rupture. 100 patients followed for 5 years. AB - We measured the configuration of the femoral condyles on lateral radiographs in 100 consecutive, prospectively-studied patients with a complete rupture of the anterior cruciate ligament, with or without associated lesions of the menisci and collateral ligaments. The patients had mainly low-to-moderate activity demands, and in all the patients the cruciate tear was primarily treated non-operatively. A quotient was calculated from the measurements of sagittal depth and axial height in order to describe the geometry of the femoral condyles. Measurements were reproducible with an intra- and interobserver coefficient of correlation of 0.89-0.98. At follow-up, after 5 (3-6) years, 16 patients had developed disability leading to reconstructive surgery. The remaining 84 patients did not have any major functional limitations, but some had reduced their activity level. Individual variations in the articular geometry were found with a more spherical shape of the femoral condyles in the patients where non-operative treatment had failed. Our findings indicate that articular geometry is of importance for function after an anterior ligament lesion. PMID- 8237328 TI - Load and length changes in an artificial ligament substitute. 10 cases of anterior cruciate ligament reconstruction. AB - In 10 patients who had reconstruction of the anterior cruciate ligament, the load and length changes in an artificial ligament substitute were measured during passive knee motion. Using a special drill guide, the ligament was placed within +/- 2 mm of the normal anatomic center on the femur. With the femoral end fixed with a bicortical screw the ligament was preloaded to 40 N at the flexion angle with the shortest intraarticular ligament length, usually 45 degrees. The change in load was then registered from 90 degrees of flexion to full extension. In 2/10 cases loads of > 200 N were registered in full extension, but the mean load was 160 N. There was a higher loss of load during the first extension/flexion cycle than during the 4th cycle. The load change correlated to the length change, but the degree of length change could not predict the maximum load level. There was a large variation in load levels between different knees, even with similar ligament placements, but the least change in load and length was obtained by an anatomic placement. Isometer readings did not predict the load level in the ligament substitute, but could indicate the angle of flexion with minimum load. Therefore, the isometer can be used to control the placement of the attachment points for the substitute. After fixation, fiber settling and stretching the ligament, as well as adaptation of the tissues, will tend to reduce the load levels. PMID- 8237329 TI - Arthroscopy of the acute traumatic knee in children. Prospective study of 138 cases. AB - We performed diagnostic arthroscopy for acute knee trauma in 138 children and adolescents aged 13 (1-15) years. The compatibility between the clinical examination and the arthroscopic findings was 59 percent. Ligament injuries were found in 32 cases, 14 of which had rupture of the anterior cruciate ligament. The compatibility in the case of ligament injuries was 31 percent. 48 patients had dislocation of the patella, and a displaced osteochondral fragment was seen in 19, 14 of which were radiographically silent. In 37 cases of distortion of the knee a correct diagnosis would have been missed unless arthroscopy had been performed. Arthroscopy is therefore indicated in children with severe distortion of the knee, hemarthrosis and with dislocation of the patella. PMID- 8237330 TI - Collagen repair not improved by fibrin adhesive. Cruciate ligament ruptures studied in dogs. AB - The anterior cruciate ligament in 30 dogs was transected and repaired by simple suture. In every other dog, fibrin adhesive (Tisseel Kit, Immuno AG, Vienna, Austria) was applied to the transection area before suturing. The proportion of organized versus unorganized and inflammatory tissue formation was assessed histologically. At 3 weeks, the amount of normal organized collagenous tissue was reduced to 20 percent both without and with fibrin adhesive. After 6 weeks, a substantial increase of organized collagenous tissue was observed after suture only, which at 12 weeks reached about 70 percent of the total area. In contrast, repair with fibrin adhesive had at 12 weeks only 30 percent of normal collagenous tissue. PMID- 8237331 TI - No effect of mop-ending on ligament healing. Rabbit studies of severed collateral knee ligaments. AB - We have tested the hypothesis that increasing the surface area of cut ligament ends by the creation of "mop-ends" may alter the mechanical properties of healing medial collateral ligaments. In one group of New Zealand white rabbits, a 4-mm midsubstance segment was removed from the right ligament creating a gap (sharp cut-end healing group). In the other group, a similar gap was created but, in addition, cut ligament ends were split longitudinally across the width of the ligament creating "mop-ends", roughly tripling the total injury surface area. In all animals, the contralateral (left) leg was not operated on and served as an internal control. At least 8 animals from each injury model were killed at 3, 6, 14, and 40 weeks post-operatively. Both histological and mechanical tests showed that sharp-cut and mop-end injuries healed at similar rates with similar material. PMID- 8237332 TI - Insertion of K-wires by hammer generates less heat. A study of drilling and hammering K-wires into bone. AB - Laboratory tests were carried out on cadaver animal bones to evaluate the thermal effect of inserting K-wires with a pneumatic hammer as compared to drilling. The mean maximum bone temperature 0.5 mm from the K-wires was 34 (22-72) degrees C using the hammer and 54 (19-100) degrees C for drilling, 1.0 mm from the K-wire the mean maximum temperature was 31 (19-52) degrees C for hammering and 47 (17 91) degrees C for drilling. The mean time for the temperature exposure in the cases of drilling was 50 s and for hammering 41 s. The hammering device may reduce the risk of heat-induced injury. PMID- 8237333 TI - Fracture of the lateral malleolus. Comparison of 2 fixation methods in cadavers. AB - 20 pairs of lateral malleolar fracture were created by an osteotome at autopsy of elderly patients and repaired on one side with staples and cerclage wire and on the other side with plate and screws. The stability of the fixation was then tested in an apparatus creating outward rotational load. The breaking load in fractures fixated with staples and wire was about 60 percent (18 SD 7.6 Nm) of the plated fibular fractures (32 SD 11 Nm). However, at an outward deflection up to 30 degrees there was no difference in breaking load with the two methods of fixation. PMID- 8237334 TI - Dislodgement of the femoral component of a cemented hip. A case report. PMID- 8237335 TI - Reduction of missed posterior dislocation of the shoulder. Report of 2 cases, 1 of them bilateral. AB - We report 2 cases of missed posterior dislocation of the shoulder, 1 of them bilateral. Prominence of the acromion and coracoid, posterior bulging and complete elimination of external rotation of the humerus lead to the diagnosis which was established by an axillary radiographic view. All 3 shoulders were treated surgically according to Mc Laughlin. At 3-4 years follow-up the function was good and there were no signs of arthrosis or avascular necrosis of the head. PMID- 8237336 TI - Lipoma arborescens with hemarthrosis of the knee. A case report. AB - Lipoma arborescens is a rare intraarticular lesion, typically with a clear yellow aspirate. We report a case of lipoma arborescens with hemarthrosis of the knee. PMID- 8237337 TI - Hydroxyapatite ceramic coating for bone implant fixation. Mechanical and histological studies in dogs. AB - The success of bone ingrowth into porous coated implants depends on several factors which can be separated into five main groups: implant related factors, such as design of implant, surface structure and pore characteristics. status of host bone bed, such as underlying disease (rheumatoid arthritis, osteoporosis), available bone stock, use of drugs and surgical technique. mechanical stabilization and loading conditions applied on the implant. adjuvant therapies such as bone grafting and HA coating which might enhance the amount of bone ingrowth. remodeling of periprosthetic bone. Once bone ingrowth has occurred, maintenance of bony anchorage depends on bone remodeling at the interface. The present series of studies were performed in order to investigate the effect of some of these factors on bone ingrowth in relation to hydroxyapatite (HA) and titanium alloy (Ti) coating when subjected to pathological and mechanical conditions mimicking the clinical situation. HA- and Ti-coated implants were inserted into the femoral condyles of mature dogs. The observation period ranged from 4 to 16 weeks, and the results were evaluated by mechanical push-out testing, histomorphometric analysis, polarized light microscopy, UV fluorescence microscopy, collagen analysis and transmission electron microscopy (microanalysis). There were no complications related to the operative procedures and all dogs were terminated according to the original time schedule. Host bone related factors were studied in the initial experiments. First, the effect of a gap between bone and implant was studied and compared with press-fit insertion. The HA-coating yielded superior effect on bone ingrowth compared to Ti in situations where the implant was surrounded by a gap and also where the implants were inserted in press-fit. Gaps of 1 mm and 2 mm around the implant were bridged by bone around HA implants whereas significantly less amounts of bone filled the gap around Ti implants. The gap-healing capacity of bone was increased even at a relatively great distance (400 microns) from the HA surface. This finding indicates that the osteoconductive effect of HA is not limited to the bone forming capacity on the surface of the implant. A positive gradient of newly formed bone was found towards the HA-coating, this gradient not being found towards the Ti-coating. In order to investigate the significance of arthritic bone changes (osteopenia) on fixation of porous coated implants we adopted the Carragheenin-induced gonarthritis model resulting in substantial bone loss as determined by CT-scanning.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8237338 TI - [Plaster of Paris: a carrier for antibiotics in the treatment of bone infections]. AB - Local antibiotherapy by diffusion from plaster of Paris is a promising method in orthopedic surgery. Nevertheless, the characteristics of this matrix have not yet been extensively studied with respect to regulation of drug-carrier capacity. From the careful comparative investigation of five commercialized plasters of Paris, it appears that the material must be constituted of fine homogeneous crystals devoid of any additive in order to obtain reproducible implants which are sufficiently hard. This condition determines their use as a biodegradable filler of bone cavities. Because of the antimicrobial spectrum, diffusion velocity and synergistic effect of sodium fusidate, amoxicillin trihydrate and sodium amoxicillin, this association of antibiotics with the plaster of Paris appears to be a useful sterilizing delivery system. The plaster of Paris allows a significant release of sodium amoxicillin during the first 3 days. This is followed by an important sustained liberation of sodium fusidate and amoxicillin trihydrate for one week. Finally, the local release of sodium fusidate for at least 2 weeks at concentrations higher than the minimum inhibition concentrations (MIC) of the most frequently infecting pathogens in osteomyelitis, will allow the sterilization of bone cavities. PMID- 8237339 TI - Tumoral calcinosis, a clinical report of eleven cases. AB - Clinical observations of 11 new cases of tumoral calcinosis are reported. The condition is characterized by calcified masses of varying size in the region of major joints. Surgical excision is recommended in selected cases determined by the size of the lesion, the deformity present and functional complaints. In this series surgical excision was done in 6 patients, and the diagnosis was confirmed histopathologically. After a complete excision of the tumor, no recurrences were seen in 5 cases with a mean follow-up time of 25 months. Incomplete excision led to multiple recurrences in one patient. PMID- 8237340 TI - Conservative treatment of acromioclavicular dislocation. Evaluation of functional and radiological results after six years follow-up. AB - Fifty-eight patients with acute acromioclavicular dislocations of Type III, IV and V (Rockwood classification) were examined to assess the late results of conservative treatment. The average age of the patients was 31 years, and the interval between injury and final review was 6.3 years. Seventy-nine percent of the patients had excellent or good late results. Surprisingly the age and activity level of the patients did not influence our late results, nor did the radiological appearance of acromioclavicular osteoarthritis or periarticular calcification. The radiological appearance of the acromioclavicular joint improved in 41% of patients. In 10 failed cases, excision of the distal end of the clavicle with reconstruction of the coracoclavicular ligament (Weaver and Dunn procedure) resulted in 90% excellent or good results after a 3-year follow up. A high percentage of excellent results can be expected after this procedure, as long as it is correctly performed. PMID- 8237341 TI - [Proximal fibulo-tibial dislocation and ischemia associated with a leg fracture. Apropos of 2 cases]. AB - Proximal fibulotibial dislocation may remain escape detection in leg injuries. Posterior dislocations are unusual and are associated with shortening of the limb due to the tibial fracture. Two cases of such dislocation are reported, associated with lower limb ischemia due to an injury of the arterial popliteal trifurcation. An anatomical model indicates the mechanism of this lesion. Such an unusual association has not be found elsewhere in literature. PMID- 8237342 TI - Orientation of the sigmoid notch of the distal radius: determination of different types of the distal radioulnar joint. AB - The orientation of the sigmoid notch of the distal radius is related to the ulnar length. In the ulna plus there is a spherical notch, in the ulna minus it is conical and in neutral it is usually cylindrical, sometimes conical. PMID- 8237343 TI - Circumscribed myositis ossificans. Report of nine cases without history of injury. AB - The clinical features of 9 patients with circumscribed myositis ossificans (CMO) are described and the effects of treatment with surgical removal of ectopic bone are assessed. The average age of these patients was 24.4 years, and the average follow-up period was 7.4 years. Early correct diagnosis remains unusual, mainly because myositis may be mistaken for bruising, sarcoma or mumps. Once histological diagnosis was established in biopsy, surgical resection of the mass was found in these series. Although spontaneous recurrence was found in this series. Although spontaneous regression of the clinical findings has been reported, we consider surgery to be necessary in CMO in order to establish the diagnosis; furthermore, when the clinical and radiological diagnosis is uncertain or when the lesion causes pain or mechanical blocking of a joint, the removal of the mass is mandatory. PMID- 8237344 TI - [Reconstitution of femoral bone stock in loosening of total hip prosthesis]. AB - Between 1986 and 1990, 18 femoral loosenings were treated for severe destruction with an original technique. The average age of the 16 patients was 62 years. Two hips had a second loosening. According to the score of Merle d'Aubigne the pain was evaluated at a mean of 2.1 preoperatively. The gait was scored 1 in half of the patients. The prostheses were trochanterodiaphyseal in 14 patients and self locking in 4 patients. The aim of this technique was: 1. to bridge the diaphyseal bone loss by healthy cortical bone, for good primary stability, 2. to reconstruct the bone stock with fragmented allografts, 3. to avoid mixing grafts and cement, by using an interposed titanium net. The frequent postoperative complications (2 fractures, 4 diaphyseal bursts, 6 false channels) occurred during extraction of the cement. Weightbearing was allowed after an average of 2 months postoperatively. With a follow-up of 33.4 months, the functional result was excellent in 11 cases (score 17 and 18). No massive resorption of the endomedullary graft was observed; trabeculation of the graft was seen in 8 patients. No migration of the prosthesis was noted. In later years, there will be a problem of extracting the rod, and there exists a risk of stress shielding, due to the length of the stem. PMID- 8237345 TI - Derotation osteotomy to correct rotational deformities of the lower extremities in children. A comparison of three methods. AB - Twenty-one supracondylar femoral and 11 tibial derotation osteotomies were performed in 17 patients to correct rotational deformities. Three different methods were used to fix the fragments after derotation: pins incorporated in plaster, plate and screws, and the Ilizarov external fixator. A retrospective review showed no difference in accuracy of derotation between the three methods. However, we recommend the use of the external fixator because it gives less malunion and permits early mobilization and (partial) weight bearing. PMID- 8237346 TI - [Carpal perilunar dislocation. Clinical study (apropos of 39 cases)]. AB - Carpal perilunar dislocations are hidden lesions with uncertain outcome and treatment which is not yet standardized. This retrospective study including 39 cases of carpal perilunar dislocations helps clarify these issues. For fresh dislocations, the diagnosis was established in 66%. Surgical treatment was performed in 26 cases. Anatomic results showed nonunion, necrosis and residual instability. Function was always decreased and was worse in open forms or after orthopedic treatment. Scaphoid screwing results were better than those from pinning. Lateral wrist x rays must always be ordered in cases of wrist trauma to avoid delayed diagnosis. Surgical treatment must systematically use a palmar incision; it permits precise repair of bones and ligamentous lesions and reduces the risk of arthritis. PMID- 8237347 TI - Quadriceps tendon rupture complicating patellar tendon release. AB - Injuries to the extensor mechanism of the knee require a profound rehabilitation. The authors present a case in which postsurgical weakening was complicated by quadriceps tendon rupture. PMID- 8237348 TI - Displaced fracture of the lunate in a child. AB - This is a report of an additional case of transverse fracture of the lunate with marked displacement, requiring open reduction and osteosynthesis. The lunate became necrotic with all features of Kienbock's disease. PMID- 8237349 TI - Vertebral aspergillosis. A case report and review of the literature. AB - A case of Aspergillus osteomyelitis of the spine is presented and the world literature is reviewed, for a total of 32 cases. Aspergillus osteomyelitis is rare, but the spine is most frequently affected. It can be induced hematogenously, by contiguity or by direct inoculation. Up to now medical treatment has been based on amphotericin B, but its toxicity is problematic. Itraconazole and fluconazole do not have this shortcoming. Additional surgery has been advocated in the past. In the present series combined medical-surgical treatment (20 cases) led to a higher survival rate (70%) than medical treatment (12 cases: 58%), but neurological recovery was much more frequent in the second group (40% versus 13%). PMID- 8237350 TI - Anomalous origin of a lumbrical muscle as cause of a trigger finger. AB - We report a case of a trigger finger due to an anomalous origin of the fourth lumbrical muscle. Total excision of the involved muscle fibers gave complete relief of symptoms. PMID- 8237351 TI - Chronic lung disease in preterm infants. AB - The increasing significance of chronic lung disease (CLD) in the neonatal intensive care unit (NICU) is a result of the increased survival of extremely small and preterm infants and a more aggressive management policy for respiratory failure. Many survive with persistent pulmonary dysfunction, the aetiology of which remains poorly understood. CLD consists of several descriptive categories which have a number of clinical features in common. PMID- 8237352 TI - Intraventricular hemorrhage in preterm neonates--a two year experience. AB - A prospective brain sonograph study was performed on 134 preterm neonates with birth weight of less than 2500 gm. Ten (7.5%) infants had intraventricular hemorrhage (IVH). The incidence of IVH in preterm neonates weighing less than 1500 gm was 13% and in neonates of less than 32 weeks gestation was 11.27%. Ninety percent of the IVH occurred within the first 72 hours of life. Prematurity, and hypoxia were considered as significant contributing factors for neonatal IVH. Six of the 7 infants with IVH grade I have a favorable neurological outcome. One of the 3 infants with IVH grade III has psychomotor delay. Routine brain sonograph study is considered practical and invaluable for diagnosis of IVH in preterm neonates. PMID- 8237353 TI - Candidemia in the neonatal intensive care unit. AB - The hospital records of 18 infants (9 males & 9 females) with one or more positive cultures for Candida species were studied retrospectively in an attempt to define the characteristics, associated factors and treatment for candidemia in the neonatal intensive care unit. The number of patients have increased recently and the mortality rate is 56% (10/18). The Candida species isolated from blood were Candida albicans in 16 cases and Candida parapsilosis in 2 cases. Fever, not doing-well, and abdominal distention were the most common presentations, prompting us to the initial impression of bacterial sepsis and/or necrotizing enterocolitis. Eleven associated factors for candidemia were relating to the measures and therapy in the neonatal intensive care unit, such as prolonged use of broad-spectrum antibiotics, parenteral hyperalimentation etc. All of these 18 patients were treated with intravenous amphotericin B. Six patients were given adequate total dose (> 25 mg/Kg), while 12 patients underwent inadequate treatment (3.7 +/- 2.9 mg/Kg). The 10 fatal patients belonged to the inadequate treatment group. As there is continual progress in neonatal intensive care units, candidemia is becoming an increasing common problem and which deserves attention. PMID- 8237354 TI - Percutaneous central venous catheterization: five year experiment in a neonatal intensive care unit. AB - We evaluated the clinical application of percutaneous central venous catheter (PCVC) in our neonatal intensive care unit (NICU). During a five-year study, 610 PCVCs were placed in 496 infants for a total of 10,243 days. The success catheterization rate was 92%. Among them, 82.7% needed only one PCVC. The body weight of babies ranged from 460 g to 5,340 g. Of 610 PCVCs, 337 (45.2%) were placed in infants weighing 1,500 g or less. About half (50.7%) of the PCVCs were placed within the first 24 hours of life. Common sites for insertion were the dorsal aspect of hands (48.8%) and the antecubital region (25.1%). Mean PCVC stay was 17.6 days (range, less than 1 to 74 days). The duration was longer (18.5 days) in infants weighing 1,500 g or less. Almost four fifth (79.2%) of 586 PCVCs were removed electively and one tenth was due to either mechanical problems (10.7%) or suspected catheter-related infection (10.1%). Of 586 PCVCs, catheter related sepsis was confirmed in 3.4% (20/586), or 2.1 per 1,000 catheter-days. No immediate complication was attributed to the insertion procedure. We conclude that PCVC is a safe and effective technique for prolonged intravenous therapy in NICU. PMID- 8237355 TI - Using quantitative ultrasound to estimate renal maturation. AB - Change in renal cortex echogenicity relative to the adjacent liver, from neonates to adults, was determined by using ultrasonography in ninety-nine cases. The kidney/liver echogenicity ratio in neonates was 0.96 +/- 0.10, decreasing with maturation to an approximate adult level (0.66 +/- 0.05) by two years of age. The kidney/liver echogenicity ratio (Y) was constructed by regression analysis on the equation Y = 0.9309-0.1201X (X = age in years) for age less than two years. PMID- 8237356 TI - [Clinical study of infants with birth weight less than 1500 grams]. AB - One hundred and seventy-seven infants of birth weight less than 1500 grams admitted to the neonatal intensive care unit of Mackay Memorial Hospital in 1987 were studied. The sex distribution, male to female ratio was 100:77, inborn 78 cases, outborn 99 cases. At one year follow-up, the mortality rate of these weighed between 500 gm and 799 gm was 100%, between 800 gm and 999 gm 54%, between 1000 gm and 1249 gm 17%, between 1250 gm and 1499 gm 19% respectively. The mortality rate of outborns was higher than that of inborns (X2 = 6.03, P < .05). The most common cause of mortality of these infants was intracranial hemorrhage, it accounts for 55% of the mortality. Seventy-three percent of the deceased cases expired during the first three hospitalization days. Of these 177 cases, 94 were put on respirator with IPPB initially, another 47 cases were on nasal CPAP. Only 36 cases didn't require respiratory therapy. Complications of the extreme prematurity and management including intraventricular hemorrhage, pulmonary hemorrhage, sepsis, pneumothorax, persistent pulmonary hypertension, disseminated intravascular coagulopathy, electrolyte imbalance, bronchopulmonary dysplasia and retinopathy of prematurity were discussed. In order to improve survival and reduce complications of these extreme prematurity, advanced monitoring system, early detection and prevention of intracranial hemorrhage, establishment of the transport system are essential. PMID- 8237357 TI - Asymptomatic reopening of ductus arteriosus after successful closure by indomethacin in premature infants. AB - Indomethacin has been demonstrated to be effective for closure of hemodynamically symptomatic patent ductus arteriosus (PDA) in premature infants. The five cases in this study had acquired asymptomatic reopening of PDA a long time after successful closure by oral indomethacin. Providing adequate oxygen, giving another course of indomethacin, preventing excessive fluid intake and keeping hematocrit level greater than 40% had been tried to close the ducti but all in vain. However, most could be expected to close spontaneously later. Herein, the possible contributing factors and the treatment rationale are discussed. PMID- 8237358 TI - Neonatal Staphylococcus aureus pancarditis: report of one case. AB - Due to poor feeding, decreased activity and severe abdominal distension, a 20-day old female infant was admitted to our hospital. The patient was diagnosed as having necrotizing enterocolitis and received a multiple segmental resection. During the hospitalization, recurrent sepsis and candidiasis occurred. On the 56th day of admission, tachycardia, thrombocytopenia and cardiomegaly were noted. Sudden onset of cardiac arrest occurred and a post-mortem examination revealed staphylococcal pancarditis. Pancarditis is a rare and fatal disease in newborns. When a patient with a catheter insertion shows unknown cause of thrombocytopenia and acute onset of cardiomegaly, an echocardiogram is highly suggested. Early diagnosis and proper parenteral antibiotics may be life-saving. PMID- 8237359 TI - Still's disease: experience in 12 children. AB - Over a 9-year period, twelve patients (8 boys, 4 girls), from 3 to 14 years old, were diagnosed as having Still's disease. Intermittent spiking high fever, poly- or pauci- articular arthritis, and typical evanescent skin rash were the most prominent clinical features. Hemogram examinations showed that 36% of the patients had anemia, ninety-two percent had neutrophilic leukocytosis and 78% had thrombocytosis. Serologically, none had positive results of rheumatoid factor and anti-nuclear antibody. Serum ferritin level was obtained from six patients and all revealed marked elevation during active disease. C-reactive protein and erythrocyte sedimentation rate were both invariably elevated. Immunologically, elevated serum concentrations of IgG, IgA, and complements (C3, C4) were found in 33%, 20%, and 17%, respectively. Furthermore, eighty percent of patients showed an increased serum level of circulating immune complexes. Aspirin (ASA) was used in all patients, but 92% of them required non-steroid antiinflammatory drugs (NSAIDs) in combination to get a better response. Sixty-seven percent of patients needed corticosteroids to control the acute systemic manifestations. Other disease-modifying agents were also used in 33% of our patients. ASA-induced liver function impairment was found in two cases. In addition, one patient experienced an episode of upper gastrointestinal bleeding. Generally speaking, the overall prognosis was good. One patient (8%) died of internal bleeding after a needle liver biopsy. PMID- 8237360 TI - Hemorrhagic shock and encephalopathy-like syndrome: report of a case with emphasis on the pathological findings. AB - The case of a 5-month-old boy with sudden onset of shock, encephalopathy, watery diarrhea, bleeding tendency, and impaired hepatic and renal functions was reported. The different pathological findings as compared with the previous reports made the term "hemorrhagic shock encephalopathy-like syndrome" (HSE-like syndrome) more suitable for this case. Marked fatty metamorphosis with hemorrhage was found with a light microscope. Electron microscopy revealed flocculent densities in the mitochondria which might be a poor prognostic indicator in this syndrome. Diffuse electron-dense deposits in the peroxisomes were also seen and the significance of this finding was discussed. The patient died on the third day after admission due to multiorgan failures in spite of our aggressive management. PMID- 8237361 TI - Intensification of relative myocardial T2-weighted magnetic resonance signals in patients with acute viral myocarditis: report of one case. AB - A 10-year-old girl was admitted to our hospital because of frequent attacks of syncope over a period of two days. A physical examination, electrocardiogram, echocardiograms and a cardiac enzyme study concluded that the girl was likely to be suffering from acute viral myocarditis. Intensities of the T2-Weighted magnetic resonance signals between the myocardium and the trapezius muscle of this patient on the first, the eighth, and the twenty-eighth hospital day were 5.1, 7.8, and 3.6 respectively. These values were significantly higher than the values of seven other patients without a history of acute viral myocarditis. Therefore, we concluded that the myocardial T2-weighted signal did intensify in patients with acute viral myocarditis. PMID- 8237362 TI - [Hemophilus aphrophilus meningitis: report of one case]. AB - Hemophilus aphrophilus, a gram negative, capnophilic slow growing bacillus, is a rarely recognized pathogen in meningitis and is most frequently seen in patients with either endocarditis or brain abscess. This article reported one case with Hemophilus aphrophilus meningitis. A 10-year-old boy presented at the emergency room with chief complaint of fever for 2 days and sudden onset of loss of consciousness. Hemophilus aphrophilus was isolated from the blood and cerebrospinal fluid. Aqueous penicillin and chloramphenicol were given for three weeks. The patient discharged without any sequelae. Three months later, fever and consciousness disturbance were noted again. No pathogen was isolated from the cerebrospinal fluid and blood culture this time, but CSF finding was consistent with bacterial meningitis. Aqueous penicillin and chloramphenicol were readministered for 30 days. The patient recovered smoothly. Because the patient had no history of CSF rhinorrhea or hypogammaglobulinemia, recurrence of the bacterial meningitis could be due to incomplete treatment during the first admission. Brain computed tomography (CT) done during the two admissions showed focal cortical enhancement in the fronto-temporo-parietal region. This is presumed to indicate infarction over these regions. The findings of brain CT are in accordance with the development of hemiplegia in the patient. It is still unknown, however, whether Hemophilus aphrophilus meningitis also causes a higher incidence of brain infarction, which was frequently noted in patients with Hemophilus influenzae meningitis. PMID- 8237363 TI - [Insulinoma in childhood--report of a case]. AB - A 14-year-old boy was having recurrent hypoglycemia over the past 6 months, so he was brought to our hospital for further evaluation. During the episodes of hypoglycemia, his serum insulin level was inappropriately high and the symptoms of hypoglycemia were dramatically ameliorated by intravenously administering glucose bolus. Computed tomography of the abdomen revealed a 1 x 1 cm mass located at the tail of the pancreas. Under the impression of an insulinoma, enucleation of the tumor was performed smoothly, and the pathological finding confirmed the diagnosis. The postoperative course was smooth and no episodes of hypoglycemia were noted after treatment. In children with hypoglycemic attacks, adequate blood sampling before correction to determine plasma insulin levels is essential. PMID- 8237364 TI - Morphological alterations of glomerulus induced by infusion of cationized ferritin. AB - Morphological alterations of the glomerulus were induced by infusion of cationized ferritin. After a direct injection of highly cationized ferritin (CF) into the left kidney of rats, endothelial injuries were followed by activation of platelets and the coagulation system after 1-2 h, which occluded capillary loops. In most glomeruli, resolving processes occurred from 2 h onward, leaving a mild thickening of the mesangial region at 7 days. On the other hand, in severely involved glomeruli, capillary loops were still obstructed even at 24 h by hypertrophic and proliferated endothelial cells as well as mesangial cells, instead of platelets and fibrin strands. After this period, exfoliation of podocytes and endothelial cells occurred over a wide area, which resulted in glomerular obsolescence at 7 days. These progressive glomerular injuries were assumed to be closely related to the persistence of CF in glomeruli, which might be caused by disturbances of glomerular clearing systems. Mild proteinuria was only noticed in severely involved cases. It is concluded that an assault on glomerular endothelial cells by cationic macro-molecules can cause thrombotic complications leading to glomerular obsolescence. PMID- 8237365 TI - Effects of sex hormones on induction of intestinal metaplasia by X-irradiation in rats. AB - The influence of sex hormones on induction of intestinal metaplasia was examined in 5 week old Crj: CD (SD) rats of both sexes. At the age of 4 weeks, the animals were gonadectomized and given testosterone or dimethyl estradiol (DES). One week after operation, they were irradiated with two 10 Gy doses of X-rays to the gastric region at a 3 day interval for a total of 20 Gy. At the termination of the experiment, 6 months after the alkaline phosphatase (ALP) positive foci in males was significantly higher than in females, in orchidectomized males or orchidectomized plus DES treated rats (P < 0.01). On the other hand, the incidence of intestinal metaplasia with ALP-positive foci in normal females appeared lower than in ovariectomized females (P < 0.01), and was increased in rats by treatment with testosterone or decreased by DES. Numbers of foci of intestinal metaplasias with Paneth cells and total numbers appeared to increase in males treated with DES. The results suggested a promoting role for testosterone in the development of ALP positive lesions and indicated considerable heterogeneity between intestinal metaplasia subtypes. PMID- 8237366 TI - An experimental model for anaplastic astrocytomas and glioblastoma using adult F344 rats and N-methyl-N-nitrosourea. AB - An experimental model for induction of gliomas corresponding to human anaplastic astrocytomas and glioblastomas is reported. Eleven week old F344 and ACI rats were given 100 or 200 p.p.m. N-methyl-N-nitrosourea (MNU) solution as their drinking water for 42 weeks. Gliomas were induced at very high incidences (82.5 92.5%) in each group. Induced gliomas showed apparent evidence of morphologic malignancy by an analysis based on diagnostic criteria of human astrocytomas. All of the gliomas from the killed animals were classified histologically into subtypes according to the classification scheme used in the diagnosis of human gliomas. The majority of macrotumors more than 1 mm in diameter in both strains were diagnosed as anaplastic astrocytomas and glioblastomas. Immunohistochemically, tumor cells in these tumors were almost negative for glial fibrillary acidic protein, while ultrastructurally neoplastic astrocytes contained glial filaments. A strain difference was observed in the ratio of histological subtypes of macrotumors. In F344 rats, astrocytic tumors diagnosed as anaplastic astrocytomas and glioblastomas of an astrocytic type formed the majority, whereas glioblastomas of mixed oligo-astrocytic type predominated in ACI rats. The results indicate that MNU-administration to adult F344 rats may provide a suitable experimental model for gliomas which occur in adult humans. PMID- 8237367 TI - Effect of PSK, a protein-bound polysaccharide preparation, on liver tumors of Syrian hamsters induced by Thorotrast injection. AB - The contrast medium Thorotrast, an agent well known to be carcinogenic, was injected into 400 congeneic Syrian hamsters. The resulting incidence of malignant hepatic tumors such as cholangiocarcinoma, hepatocellular carcinoma and hemangiosarcoma, was significantly higher in the male experimental group than in the control group, and the 50% survival period in the male group was shortened by about 100 days (P < 0.01). However administration of the antitumor drug PSK (Polysaccharide Kureha), a protein bound-polysaccharide extracted from basidiomycete fungi, prevented this carcinogenic effect. The incidence of malignant hepatic tumors in the experimental group was 22.5% compared with 2.8% in the control group (P < 0.01) and 10.5% in the PSK-treated group (P < 0.01). PSK also increased the 50% survival period by 61 days (P < 0.01). PMID- 8237368 TI - Porcine vascular smooth muscle cells immortalized with SV40 ori-defective DNA: characteristics of cell growth and collagen synthesis. AB - A cell line derived from medial smooth muscle cells (SMC) was established from the porcine coronary artery by transfection with ori-defective simian virus 40 plasmid DNA (SV40 DNA). The characteristics of transfected cells (SV40-SMC) such as cell growth, collagen and non-collagen syntheses were investigated. SV40-SMC expressed SV40 large T antigen, c-myc and c-myb encoded proteins in the nuclei. SV40-SMC demonstrated a 'hills and valleys'-like arrangement in overconfluence and actin filaments upon immunofluorescent staining. Under electron microscopic observation, SV40-SMC had larger amounts of synthetic organelles and smaller amounts of filament bundles than those of SMC. SV40-SMC demonstrated three times higher growth activity and 4.4 times greater cellular density than SMC. Smooth muscle cells did not grow in media containing 5% plasma derived serum (PDS) instead of normal serum, whereas SV40-SMC proliferated in this medium. SV40-SMC did not grow in soft agar gel, while HeLa S3 cells, a cell line of human cervical carcinoma, formed colonies in this gel. By immunofluorescent (IF) staining, collagen phenotypes I, III, IV and V were detected in both SV40-SMC and SMC. However, protein synthesis including collagen and non-collagen was higher in SV40 SMC than in the control sample. It was concluded that SV40-SMC were a continuous cell line for vascular SMC regarding morphological characteristics, and demonstrated a higher growth activity, with increased collagen and non-collagen syntheses. This cell line is useful for the investigation of atherogenesis in relation to a proliferation of SMC and an accumulation of extracellular matrices in vascular intima. PMID- 8237369 TI - Markers of surface mucous cell type human gastric cancer cells: galactose oxidase Schiff reactive mucins, monoclonal antibody SH-9 reactive mucins and cathepsin E. AB - Cellular differentiation of gastric cancer cells allows the classification of cell type into surface mucous cell, pyloric gland cell, intestinal absorptive cell and goblet cell types by mucin histochemistry and pepsinogen (Pg) immunohistochemistry. Surface mucous cell differentiation of gastric cancers of each histologic type has previously been detected by the galactose oxidase-Schiff (GOS) reaction although this is not always positive in all cases. Mucus granules of surface mucous cells of normal gastric mucosa show an intense reactivity for SH-9 (monoclonal antibody against CA125-bearing antigenic molecule fragments). Cathepsin E is also expressed in the cytoplasm of surface mucous cells, weakly in absorptive cells of duodenal villi and occasionally in pyloric gland cells. Expression of SH-9 reactive mucin and of cathepsin E were therefore investigated as possible additional markers to distinguish between the gastric cancer cell type in 203 primary stomach cancers. SH-9 reactive mucin was found selectively in GOS positive cancer cells of surface mucous cell type and/or cancer cells unclassified by mucin histochemistry. These latter cells were therefore classified into the surface mucous cell category. Cathepsin E was found mainly in cancer cells of the GOS positive surface mucous cell type and occasionally, in intestinal absorptive and pyloric gland cell types. Galactose oxidase-Schiff, SH 9 and cathepsin E reactive or positive cancer cells were found in 145 (71.4%), 151 (74.4%) and 144 (70.9%), respectively, of the 203 primary stomach cancers investigated. PMID- 8237370 TI - Sclerosing hemangioma of the lung: pathological study and enzyme immunoassay for estrogen and progesterone receptors. AB - Sclerosing hemangioma of the lung is a rare benign neoplasm of uncertain histogenesis. In this study, thirteen cases of sclerosing hemangiomas were analyzed by immunohistochemistry for various histochemical markers including surfactant apoprotein. The pale cells were shown to share several immunohistochemical markers with epithelial lining cells and were suggested to be immature cells differentiating toward epithelial lining cells. Electron microscopic study failed to characterize the exact nature of pale cells, but it disclosed the precise structure of large lamellar bodies, which are frequently present in the cystic spaces of sclerosing hemangioma. These large lamellar bodies had common morphological characteristics to lamellar inclusion bodies of type II pneumocytes or epithelial lining cells. Eight cases of sclerosing hemangioma obtained freshly at surgery had a quantitative analysis of estrogen and progesterone receptors performed by enzyme immunoassay. The value of estrogen receptors ranged from 0 to 7.8 fmol/mg protein. In addition to this quantitative analysis, the immunohistochemical study demonstrated estrogen receptors mainly in the pale cell nuclei of several sclerosing hemangiomas. Estrogen receptors may play an important role in growth, differentiation and surfactant production in the sclerosing hemangioma. PMID- 8237371 TI - Encapsulated papillary carcinoma of the thyroid gland: clinicopathological and cytofluorometric study in comparison with non-encapsulated papillary carcinoma. AB - Clinicopathological and cytofluorometric studies were performed on nine encapsulated (EPC) and 23 non-encapsulated (non-EPC) carcinomas of the thyroid gland. The average age of the patients with EPC was 33 years, which was significantly younger than that of those with non-EPC. The average tumor size of EPC was twice as large as that of intraglandular non-encapsulated carcinoma. All patients with EPC were alive without disease, but three out of 23 patients with non-EPC had a recurrence of the disease or died. Cytofluorometric studies showed that the mean nuclear DNA content and percentage of tumor cells in the S-G2M phase of EPC were lower than that of non-EPC. According to the DNA histogram pattern, all EPC showed distinct modal DNA values in the diploid or near diploid region of normal cells. However non-EPC, especially extra-glandular non encapsulated papillary carcinoma, showed a wide variety of DNA histogram patterns. The present study suggested that EPC is a distinct subtype of papillary carcinoma of the thyroid gland clinicopathologically and that cytofluorometrically it is different from non-EPC. PMID- 8237372 TI - A case of severe IgA nephropathy associated with psoriatic arthritis and idiopathic interstitial pneumonia. AB - A patient is described with severe IgA nephropathy associated with psoriatic arthritis, idiopathic interstitial pneumonia and brain hemorrhage that developed serially over one and a half years. The histological findings of the renal biopsy showed severe endo- and extracapillary proliferative glomerulonephritis. Massive IgA deposits were observed by immunofluorescence not only in the mesangium but also along the capillary walls. Electron microscopy revealed abundant electron dense deposits in the mesangial and subendothelial areas. The overlapping or coexistence of these conditions has rarely been reported. PMID- 8237373 TI - Spindle cell hemangioendothelioma: a report of two cases. AB - Two cases of spindle cell hemangioendothelioma (SCH) are reported. One of the patients was a 16 year old Japanese female, who had been suffering from Ollier's disease (multiple enchondromatosis) since 3 years of age and had developed multiple SCH in the right leg at the age of 11 years. Spindle cell hemangioendothelioma lesions coincided with the site of enchondromatosis and increased in number thereafter. This is the first report of Ollier's disease complicated with multiple SCH. Another patient, a 33 year old Japanese female, who was a carrier of hepatitis B virus (HBV), developed solitary SCH in the lateral aspect of the right ankle where a lipoma was extirpated 10 years previously. Tumor cells of both cases were composed of four cell types: (i) spindle cells; (ii) epithelioid cells; (iii) vacuolated endothelial cells; and (iv) usual endothelial cells. Endothelia in the cavernous area and vacuolated cells reacted to Ulex europaeus agglutin 1 (UEA-I), factor VIII-related antigen and vimentin. Spindle cells and epithelioid cells reacted only to vimentin. PMID- 8237374 TI - [Effect of intracerebroventricular injection of N-methyl-D-aspartate on learning and memory in mice]. AB - In one trial passive avoidance response in mice, the effect of N-methyl-D aspartate (NMDA) on acquisition, consolidation and retrieval of memory were observed after icv administration of 10 ng, 1 ng or 0.1 ng/mouse. The results showed that NMDA 10 ng/mouse could promote the consolidation of memory in step down test and 1 ng/mouse could promote the retrieval of memory in step-through test. NMDA 1 ng/mouse significantly improved the retrieval impairment induced by 30% alcohol and the consolidation impairment induced by NaNO2 in step-down and step-through tests. The effect of NMDA on memory was antagonized by NMDA receptor antagonist, AP5. These results provide direct evidence for the action of NMDA subtype of glutamate receptor on memory. PMID- 8237375 TI - [The cytotoxicity and action mechanism of ranunculin in vitro]. AB - This paper describes the cytotoxicity of ranunculin (RAN) and its mechanism of action. The IC50 of RAN against the KB and Bel7402 cells in colony test were found to be 0.21 and 0.35 mumol/L respectively. RAN inhibited the incorporation of 3H-labeled precursors into DNA and RNA of L1210 cells. RAN (15 mumol/L) markedly decreased DNA synthesis catalyzed by DNA polymerase I and promoted the generation of superoxide anions in DMSO/KO2 system. In the meantime, SOD and CAT were shown to partly revoke the inhibitory effects of RAN upon the incorporation of 3H-TdR into DNA. No direct reaction between RAN and DNA template was observed and no effect of RAN on DNA TOPO II or RNA polymerase was found. Our results suggest that the cytotoxicity of RAN in vitro may be due to inhibition of DNA polymerase and increase of oxygen free radicals. PMID- 8237376 TI - [Effects of six naphthalenesulfonamide derivatives on LPS-induced release of tumor necrosis factor from mouse peritoneal macrophages]. AB - The effects of six naphthalenesulfonamide derivatives were studied on the LPS induced release of tumor necrosis factor (TNF) from mouse peritoneal macrophages primed with A23187. The calmodulin (CaM) antagonist, N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide (W-7) and its derivatives N-(6-aminobutyl)-5-chloro-1 naphthalenesulfonamide and N-(6-aminoethyl)-5-chloro-1-naphthalenesulfonamide (10 400 ng/ml) were found to inhibit LPS-induced TNF release in a dose-dependent manner, and the protein kinase C (PKC) activator, N-(n-heptyl)-5-chloro-1 naphthalenesulfonamide (SC-10) and its two derivatives, N-(n-quinyl)-5-chloro-1 naphthalenesulfonamide and N-(n-butyl)-5-chloro-1-naphthalenesulfonamide (1-16 micrograms/ml) were shown to increase LPS-induced TNF release at suboptimal doses in a dose-dependent manner. These results suggest that the LPS-induced release of TNF is CaM-dependent and PKC may play an important role in this process. PMID- 8237377 TI - [HPLC--detection of hydroxyl radicals in striatum extracellular fluid in rats subjected to reperfusion after cerebral ischemia and the action of vitamin E]. AB - The formation of hydroxyl radical in the extracellular fluid in striatum of rats subjected to 4-vessel occlusion during ischemia and reperfusion was measured by HPLC with UV detector using salicylic acid as a trapping agent. Hydroxyl radicals react with salicylic acid yielding 2,3- and 2,5-dihydroxybenzoic acids which can be separated by liquid chromatography. The striatum was perfused with artificial cerebrospinal fluid containing 0.5 mmol.L-1 salicylic acid at a flow rate of 2.5 microliters.min-1. The results indicate that the concentration of 2,5 dihydroxybenzoic acid in the perfusates was highest during 25 min reperfusion and no change was found during ischemia. Vit E at 30, 60 and 120 mg.kg-1 i.p. 20 min before ischemia was shown to dose dependently suppress the rise of 2,5 dihydroxybenzoic acid after reperfusion. The results suggest that the HPLC method is useful and reliable for the measurement of hydroxy radical in global ischemia in vivo. PMID- 8237378 TI - [Pharmacokinetics of dihydroqinghaosu in human volunteers and comparison with qinghaosu]. AB - Qinghaosu (QHS), also known as artemisinin and arteannuin, is a novel type of sesquiterpene with a peroxide linkage isolated from the Chinese herb Artemisia annua L. Since its discovery as an antimalarial with low toxicity, hundreds of derivatives have been synthesized among them artesunate (ATS), artemether (ATM) and dihydroqinghaosu (DHQHS) were found to be more active than QHS itself. A suppository of QHS, a dual-pack dosage form of ATS (artesunic acid to be dissolved in sodium bicarbonate solution just before iv injection) and an oil solution of ATM for im injection had been approved by our Ministry of Health for clinical use. However, a preparation for oral administration is still not available. We have reported that when dogs were given QHS tablets orally at the dose of 70 mg/kg, no drug was detected in the serum using the RIA method, whereas appreciable serum concentration was found by the same method when dogs were given DHQHS tablets at a dose as low as 10 mg/kg. This paper reports the pharmacokinetics of DHQHS in man studied with the RIA method and compared with QHS. When DHQHS in tablet form was given to human volunteers at doses of 1.1-2.2 mg/kg, peak serum levels of 0.13-0.71 micrograms/ml were obtained in 1.33 h with MRT of 2.26-2.36 h. When QHS tablets were given at the dose as high as 15 mg/kg, however, the peak serum level found in 1.5 h was only 0.09 microgram/ml with MRT of 1.33 h. Therefore, the bioavailability of QHS tablets is only 1.62-10.08% that of DHQHS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237379 TI - [Synthesis of gona-3,5-dienes and studies on their activity for interruption of early pregnancy]. AB - A series of gona-3,5-dienes were synthesized from norgestrel, norethisterone 17 alpha-hydroxyprogesterone and ethisterone via reduction of alpha, beta unsaturated 3-ketones to allylic alcohols and dehydration with concomitant migration of the double bond. The structures of these gona-3,5-dienes were verified by MS and 1H-NMR spectral data. Pharmacological results showed that compound IVb2 in this series gave rise to significant interruption of early pregnancy in mice. The mechanism of the early pregnancy terminating effect of this compound was discussed. PMID- 8237380 TI - [Isolation and identification of two new saponins from Bupleurum smithii Wolff]. AB - Two new saikosaponins of oleanane type were isolated from the roots of Bupleurum smithii Wolff, collected in Minhe County, Qinghai, namely saikosaponin k (VIb) and saikosaponin 1 (VIII). Their structures were elucidated on the basis of spectral analysis of UV, IR, 1HNMR, 13CNMR and FAB-MS. Saikosaponin k (VIb) was identified as 3 beta, 16 beta, 23,28-tetrahydroxyoleana-11,13(18)-dien-3-O-beta-D xylopyranosyl-(1-->2)-beta- D-glucopyranosyl-(1-->3)-beta-D-fucopyranoside, and saikosaponin 1 (VIII) as 3 beta, 16 alpha, 23,28,30-pentahydroxyoleana-11,13(18) dien-3-O-beta-D- glucopyranosyl-(1-->3)-beta-D-fucopyranoside, respectively. PMID- 8237381 TI - [Structure determination of saponin XI, XII and XIII from Dipsacus asper Wall]. AB - Three new triterpenoid glycosides (XI, XII and XIII) were isolated from the ethanol extract of the roots of Dipsacus asper Wall. Their structures were deduced as 3-O-[beta-D-glucopyranosyl(1-->4)][alpha-L-rhamnopyranosyl (1-->3)]- beta-D-glucopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-alpha- arabinopyranosyl-hederagenin (XI), 28-O-beta-D-glucopyranosyl(1-->6)-beta-D glucopyranosyl ester of XI (XII) and 3-O-[beta-D-xylopyranosyl(1-->4)-beta-D glucopyranosyl (1-->4)][alpha-L-rhamnopyranosyl(1-->3)]-beta-D-glucopyranosyl(1- >3)- alpha-L-rhamnopyranosyl(1-->2)-alpha-arabinopyranosyl-oleanolic.ac id 28-O beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl ester (XIII), respectively, based on spectral and chemical analysis. PMID- 8237382 TI - [Studies on chemical constituents of Gynostemma compressum]. AB - Four new saponins, gycomoside I-IV, were isolated from the aerial part of Gynostemma compressum X.X. Chen et D.R. Liang. The structures were elucidated on the basis of spectral and chemical evidences as 1 beta,3 beta,12 beta,20(S),26 pentahydroxy-dammar- 24(25)-en-20(S)-O-beta-D-glucopyranosyl1-6--beta-D glucopyranoside (gycomoside I); 1 beta,3 beta,12 beta,20(S)-tetrahydroxy-dammar 24(25)-en-3-O- beta-D-glucopyranosyl-20(S)-O-beta-D-glucopyranosyl1-6--beta-D-glu copyranoside (gycomoside II); 1 beta,3 beta,12 beta,20(S), 26-pentahydroxy-dammar 24(25)-en-20(S)-O- beta-D-glucopyranoside (gycomoside III) and 1 beta,3 beta,12 beta,20(S),26-pentahydroxy-dammar-24(25)- en-3-O-beta-D-glucopyranosyl-20(S)-O beta-D-glucopyranosyl1-6--bet a-D-glucopyranoside (gycomoside IV). PMID- 8237383 TI - [PAF antagonistic benzofuran neolignans from Piper kadsura]. AB - In a continuing search for PAF antagonists, five benzofuran neolignans have been isolated from the aerial part of Piper kadsura (Choisy) Ohwi, a Chinese traditional drug used for the treatment of inflammation and rheumatic conditions. The structure determination was based upon spectroscopic analysis. Two of the neolignans were found to have new structures and were named as (-)-denudatin B (the enantiomer of denudatin B, II) and kadsurenin M (7S,8S-3,4,3'-trimethoxy-7' oxo-nor-8',9'-7.O. 4',8,5'-neolignan, V). The known compounds kadsurenon (I), (-) acuminatin(III) and (+)-licarin A(IV) were also obtained from the same source. ( )-Denudatin B (II) showed potent PAF antagonistic activity in 3H-PAF receptor binding assay. PMID- 8237384 TI - [Column switching HPLC method for determination of dextrorphan, an active metabolite of dextromethorphan, in plasma]. AB - An HPLC method for the determination of dextrorphan, an active metabolite of dextromethorphan, in plasma was established using column switching technique. The column switching system was equipped with a per-column of 30 mm x 5 mm ID, packed with mu Bondapak C18, 37-50 microns, and an analytical column of 150 mm x 5 mm ID, packed with YWG-C18, 5 microns. A 0.2% acetic acid solution was used as the pretreating mobile phase to wash out impurities from the per-column. The analytical mobile phase consisted of acetonitrile-water-acetic acid-triethylamine dichloromethane (17:82:1:0.05:0.025). The plasma samples were directly injected into the HPLC system after enzymatic hydrolysis of dextrorphan glucuronide ester conjugate to free form with beta-glucuronidase. The dextrorphan was monitored with a fluorescence detector at 290 nm (excitation) and 315 nm (emission). The method was linear within the plasma concentration range of 20-640 ng/ml (r = 0.9987), and the detection limit was 4 ng/ml. The mean recoveries of the method averaged 103.8%. The relative standard deviations of the assay were less than 10% for both within-day and between-days. PMID- 8237385 TI - [Analysis of tramadol and its metabolites in human urine]. AB - A GC-MS method for the analysis of tramadol and its four metabolites in human urine is described. The urine samples were acid hydrolyzed with hydrochloric acid, cleaned with diethyl ether and extracted with dichloromethane-isopropanol (9:1). After derivatization, the solution was analyzed with GC-MSD. Tramadol and its 4 metabolites were detected in urine samples 2-40 hours after oral administration. The recovery of tramadol was 85.2% +/- 5.4 (n = 3), the detection limit was down to 12.5 pg. The derivatization methods was discussed. PMID- 8237386 TI - [Study on sodium norcantharidate albumin microspheres]. AB - In this paper sodium norcantharidate (Na2NC) was chosen as model drug and sodium norcantharidate albumin microspheres (Na2NC AM) was prepared by an optimal procedure. The embedding nature, appearance, morphology, size and size distribution, release characteristics in vitro, freeze-drying, 60Co radiosterilization and stability of Na2NC AM were tested. The results showed that the mean size was 0.43 +/- 0.12 micron, embedding concentration was 20.34-26.75 micrograms/mg, embedding ratio was 21.9-26.4%, release characteristics in vitro was in accord with Higuchi equation. Freeze-drying and 60Co radiosterilization showed no influence on Na2NC AM. The stability of Na2NC AM was good after 3 months storage. PMID- 8237387 TI - [Study on the degradation kinetics of vitamin K3 powder]. AB - The effect of temperature and humidity on the rate of discoloration and degradation of vitamin K3 powder was studied via accelerated test under isothermal, constant humidity and dark conditions. The photodegradation of vitamin K3 powder was studied under room temperature and constant humidity. The changes of color and concentration of vitamin K3 were monitored by diffuse reflectance spectroscopy and high performance liquid chromatography, respectively. At temperatures 50-90 degrees C and relative humidities 50-93%, the rate equation of discoloration is -dR/dt = KRR, that of degradation is d(c(o) c)/dt = kc (c(o)-c)1/2, and that of photodegradation is -dR/dt = kp (Ro-R)-1, where Ro and R are initial relative reflectance and relative reflectance at time t, respectively, c(o) and c are initial concentration and concentration at time t, respectively, kR, kc and kP are apparent rate constants of discoloration, degradation and photodegradation, respectively. The general equation of the relationship between the apparent rate constant k, absolute temperature T and relative humidity RH is k = k' exp (-Ea/RT) exp (m' RH), where Ea is apparent activation energy, k' and m' are constants. Within the ranges of experimental temperatures and humidities, the average apparent activation energies of discoloration and degradation are 105.51 and 130.72 kJ/mol, respectively. PMID- 8237388 TI - [Synthesis and antitumor activity of beta-diketonato complexes of titanium]. AB - Nine cis-beta-diketonato titanium (IV) complexes were synthesized and their chemical structures were confirmed by IR, UV spectra and elemental analyses. Among them 6 complexes have not been reported before. Preliminary pharmacological experiments showed that this kind of complexes were active against tumors. PMID- 8237389 TI - Pharmacokinetics and relative bioavailability of tablet of micronized glibenclamide in 4 Chinese healthy men. AB - Pharmacokinetic characteristics and relative bioavailability of the regular preparation (RG) and micronized preparation (MG) of glibenclamide (Gli) were studied in 4 Chinese healthy men. Each volunteer entered 2 consecutive experiments at the same dose (10.5 mg) of RG and MG tablets given orally. Blood samples were drawn before and 0.5, 1, 2, 3, 4, 6, 8, 10, and 12 h after po. A selective HPLC method with a uv spectrophotometric detector (sensitivity: 16 ng.ml-1) was established for determining serum Gli concentration. Cmax for RG was 212 +/- 41 ng.ml-1, for MG 529 +/- 73 ng.ml-1. Tmax for RG and MG were 3.5 +/- 0.6 and 2.2 +/- 0.3 h, respectively. The relative oral bioavailability of MG was found to be 77%, increased approximately 1.7-fold that of RG. The other pharmacokinetic parameters (Vd, Cl, T1/2) were substantially the same after ingestion of GR and MG. PMID- 8237390 TI - Effects of lisinopril and captopril on calcium in rat heart. AB - We studied the effects of lisinopril (Lis) and captopril (Cap), two angiotensin converting enzyme inhibitors, on calcium in ischemia/reperfusion and normal rat hearts. Ischemia/reperfusion hearts were subjected to 15 min ischemia followed by 1 or 30 min reperfusion. Lis 0.1 mumol.L-1 and Cap 200 mumol.L-1 decreased the concentration of calcium in ischemia/reperfusion hearts (the content of calcium in reperfusion 1 min heart were reduced from 4.0 +/- 0.6 to 2.7 +/- 0.5 and 3.0 +/- 0.9 mumol/g dry wt respectively). In cultured cell of neonatal rat heart, both drugs inhibited the uptake of 45Ca2+. The activity of Na+,K(+)-ATPase prepared from rat heart was increased (activity increased from 15.7 +/- 2.3 in control group to 21.2 +/- 2.0 and 22.0 +/- 3.1 mumol/h mg protein in Lis and Cap groups, respectively). This calcium lowering effects of Lis and Cap may be important in protecting the ischemia/reperfusion damage of myocardium. PMID- 8237391 TI - Inhibition of formalin-induced responses of spinal dorsal horn neurons by stimulation of lateral reticular nucleus in rats. AB - The effects of stimulation of the lateral reticular nucleus (LRN) on responses of spinal dorsal horn neurons produced by formalin (5%, 50 microliters, sc) were investigated. In 12 out of 19 neurons tested, formalin induced 2 distinct phases of response. The first phase was initiated immediately after the injection of formalin and lasted for 3-8 min. The second phase started gradually 20-35 min after administration of formalin and lasted 30-65 min. Both phases of the response were inhibited by LRN stimulation. PMID- 8237392 TI - Protective effects of 3,6-dimethylamino-dibenzopyriodonium edetate on global ischemia reperfused isolated rat hearts. AB - The effects of 3,6-dimethylamino-dibenzopyriodonium edetate (IHC-72) on global ischemia reperfused rat hearts were investigated. In the isolated working rat heart, 40-min global ischemia followed by 30-min reperfusion resulted in increases of ventricular tachycardia (VT) and ventricular fibrillation (VF), increases of creatine kinase (CK) release and malondialdehyde (MDA) contents, but decreased superoxide dismutase (SOD) activity. Following ischemia and reperfusion, the accumulation of myocardial calcium increased. IHC-72 50 mumol.L 1 given 10 min before ischemia and during reperfusion decreased the cardiac CK release, VT, and VF, reduced the MDA contents, prevented the reduction of SOD activity and attenuated the accumulation of myocardial calcium and sodium vs control. These results indicated that IHC-72 protected myocardial reperfused injury. PMID- 8237393 TI - Effect of phencyclidine on contraction of porcine coronary vessel strips. AB - Classic muscular contraction experiment was used to study the effect of phencyclidine (Phe) on spiral strips of porcine coronary vessels. It showed that Phe and its analogs: 1-[1-(2-thienyl)cyclohexyl] piperidine (TCP), (+)-5-methyl 10, 11-dihydro-5H-dibenzo [a,d] cyclohepten-5, 10-imine maleatel (dizocilpine maleate) and sigma receptor ligand N-allyl-N-normetazocine (SKF10047) all exhibited a concentration-dependent contraction of porcine coronary vessel strips. Phe had no effect on the electrically stimulated contraction of the spiral strips. Dextromethorphan (Dex), a Phe receptor antagonist, and haloperidol (Hal), a sigma receptor antagonist, partially antagonized and suppressed the actions of Phe in a non-competitive manner. These results suggested that Phe, TCP, dizocilpine maleate, and (+), (-)-SKF10047 cause contraction through Phe and sigma opioid receptors. PMID- 8237394 TI - Lytic solution of platelet-rich plasma clots potentiated platelet response to agonists by action on Ca2+ homeostasis. AB - Effects of thrombolytic agents and lytic solution of platelet-rich plasma (PRP) clots (LSPC) on platelet activation as indicated by platelet aggregation, generation of malondialdehyde (MDA), and the concentration of intracellular free calcium ([Ca2+]i) in rats were investigated. Neither urokinase nor streptokinase in vitro showed adverse effects on platelet function. The solution of PRP clots incubated either alone (as control) or with urokinase or streptokinase 2000 IU.ml 1 at 37 degrees C for 45 min potentiated the increase of platelet aggregation and MDA formation and produced a persistently high level of [Ca2+]i stimulated by thrombin, and platelet aggregation induced by ADP. But LSPC had no effects on the Ca2+ influx or the release of intracellular stored Ca2+, and no significant difference was found in the promotion of platelet response to agonists between the solution of the clots warmed in the presence or absence of thrombolytic agents. In the thrombosis model in rat abdominal aorta, both urokinase and streptokinase (40,000 IU.kg-1) slightly inhibited electrically stimulated thrombosis. In contrast, LSPC (600 microliters.kg-1) considerably enhanced the thrombosis. These findings suggested that the changes of platelet function in ischemic patients receiving thrombolytic therapy may be mediated by the proteolytic products of clots through acting on [Ca2+]i homeostasis after platelet stimulation rather than by the thrombolytic agents per se. PMID- 8237395 TI - Second peak of plasma diazepam concentration and enterogastric circulation. AB - Intragastric food administration caused a pronounced second peak of plasma diazepam concentration in rabbits after iv diazepam 5 mg.kg-1. The second peak disappeared after gastrostomy and choledochostomy. A large amount of diazepam was found in the gastric juice while its content in bile remained at a much lower level during the whole experiment. These results suggested that diazepam may undergo an enterogastric circulation in addition to its enterohepatic circulation, with the former mainly contributing to the appearance of the second peak. PMID- 8237396 TI - Effects of 4-[4''-(2'',2'',6'',6''-tetramethyl-1''-piperidinyloxy)amino]-4'-de methylepipodophyllotoxin on immune function in mice. AB - 4-[4''-(2'',2'',6'',6''-Tetramethyl-1''-piperidinyloxy)amino]-4'- demethylepipodophyllotoxin (GP-7) 10-40 mg.kg-1 ip daily for 7 d reduced the specific antibody formation of splenocytes, serum agglutinin titer, and hemolysin HC50 in mice immunized with SRBC. GP-7 inhibited the footpad delayed hypersensitivity reaction and decreased the weights of spleen and thymus, but did not affect the phagocytic function of the peritoneal macrophages. In vitro the proliferation of mouse splenic lymphocytes activated by Con A was markedly inhibited by GP-7 in a concentration- dependent manner. At concentrations of 0.05 5 mg.L-1, the inhibition rates were 24-96%. These results suggested that GP-7 was an immunosuppressive agent. PMID- 8237397 TI - Endocardial endothelium modulates cardiac responses to histamine and impromidine in isolated working right ventricle of guinea pigs. AB - In isolated working right ventricle of guinea pigs, the hypothesis that endocardial endothelium (EE) might be involved in the modulation of cardiac responses to histamine receptor agonists was tested. The functional EE was denuded by switching Krebs perfusion solution to the solution containing saponin (30 micrograms.ml-1) for 2 min at a rate of 16 ml.min-1, followed by thorough washing with Krebs solution. The cardiac responses to histamine receptor agonists were compared in the presence and absence of EE. Bolus injection of histamine 0.5 mg into right ventricle elevated the right ventricular pressure (RVP), +dP/dtmax, and -dP/dtmax by 11%, 17%, and 35%, respectively, in the presence of intact EE; whereas by 30%, 43%, and 92%, respectively, after chemically selective denudation of EE with saponin. Similarly, impromidine (a H2 receptor agonist) 1, 3, 9, 27, and 54 micrograms obviously potentiated the RVP and +/- dP/dtmax in a concentration-dependent manner in preparations either with or without EE. The effects, however, could be greatly enhanced in the absence of EE. Pulmonary outflow was declined at 27 micrograms impromidine in EE-removed group. The results suggested that the augmentation of cardiac responses produced by histamine receptor agonists in EE-denuded preparations might be due to blockade of release of endothelium-derived relaxing factor, resulting in a reduced abbreviating effect on the myocardial contraction. PMID- 8237398 TI - Pharmacokinetics of sustained-release tablets of metoprolol in Chinese. AB - Single and repeated oral doses pharmacokinetic studies of metoprolol sustained release tablets (Sino-Swed Pharmaceutical Co Ltd) were performed on 12 Chinese healthy subjects in an open randomized crossover manner, using metoprolol tablets from Sweden Astra International Pharmaceutical Co Ltd as control. Drug concentrations in plasma were assayed by gas chromatography-electron-capture detector method. The percent of drug absorbed in vivo at 1, 2, 4, 6, 8 h correlated well with the amount of drug released in vitro at corresponding time (P > 0.05). Pharmacokinetic parameters in Chinese after metoprolol tablets were comparable to the reported data in foreigners. PMID- 8237399 TI - Effects of (-)-stepholidine and tetrahydroberberine on high potassium-evoked contraction and calcium influx in rat artery. AB - The relaxant effects of (-)-stepholidine ((-)-SPD) and tetrahydroberberine (THB) on rat aorta were studied in vitro. (-)-SPD IC50 18.1 (95% confidence limits 11.1 29.5) mumol.L-1 and THB IC50 18.6 (95% confidence limits 9.2-37.9) mumol.L-1 inhibited the contractions caused by KCl (100 mmol.L-1) concentration dependently. Both (-)-SPD and THB markedly inhibited the 160 mmol.L-1 KCl stimulated 45Ca influx. The inhibitions by (-)-SPD 10 mumol.L-1 and 100 mumol.L-1 were 18 +/- 13% (P > 0.05) and 47.0 +/- 2.8% (P < 0.01), respectively. The inhibitions by THB 10 mumol.L-1 and 100 mumol.L-1 were 36 +/- 9% (P < 0.01) and 43 +/- 8% (P < 0.05), respectively. The results showed that the effective concentrations of the 2 drugs inhibiting high KCl-induced contraction and 45Ca transmembrane influx in rat thoracic aorta were at a similar level, and that they were nearly 1/100 and 1/10 of those of verapamil respectively, indicating that ( )-SPD and THB had similar calcium channel blocking effect on rat artery, but were weaker than verapamil. PMID- 8237400 TI - Effects of triptonide on mouse immune functions. AB - Triptonide (Tri) extracted from Tripterygium wilfordii Hook inhibited the proliferation of mouse splenocytes induced by suboptimal concentration of concanavalin A or lipopolysaccharide at concentrations of 0.02, 0.1, and 0.5 microgram.ml-1. It showed a suppressive effect on one way mixed lymphocyte culture (MLC) at 0.1-0.4 microgram.ml-1. The lymphocytes harvested from the first Tri (0.4 microgram.ml-1)-containing MLC inhibited the second MLC after being washed and irradiated with 60Co source (30 Gy). Serum anti-sheep red blood cell antibody (hemolysin) formation and clearance of charcoal particles were also suppressed by Tri in vivo. Although delayed hypersensitivity (DH) reaction to dinitrofluorobenzene (DNFB) was inhibited by Tri 1.2, 2.5, and 5.0 mg.kg-1 (ip, qd x 5 d), the spleen cell interleukin-2 secretion activities of these mice were not suppressed. Graft vs host reaction (GVHR) was not inhibited by Tri 2.5 and 5.0 mg.kg-1 (ig, qd x 5 d). Helper T cells (Th)/suppressor T cells (Ts) ratio was reduced at 2.5 mg.kg-1. The effects of Tri on mouse thymus and spleen weight were related to the age. Tri (1.2, 2.5, 5.0 mg.kg-1) had no effect on thymus and spleen weights in 8-wk-old mice. However, it increased the thymus and spleen weights in 12-wk-old mice at doses of 1.2 and 2.5 mg.kg-1. The data indicated that Tri had extensive suppressive effects on mouse immune function and its mechanism may be related to the reduction of Th/Ts ratio and the induction of Ts cells. PMID- 8237401 TI - Effects of Phytolacca acinosa polysaccharides I on immune function in mice. AB - Radioactivities of [3H]TdR uptaken by splenocytes and released from [3H]TdR labeled YAC-1 cell line were measured to determine the degree of lymphocyte proliferation and natural killer (NK) cell activity. Seven days after mice treated with Phytolacca acinosa polysaccharides I (PAP-I) 5-50 mg.kg-1, the NK cell activity, and lymphocyte proliferation induced by Con A 5 micrograms.ml-1 or lipopolysaccharides 10 micrograms.ml-1 were significantly augmented. Splenocytes from mice treated with ip PAP-I 5-50 mg.kg-1 were incubated with Con A 5 micrograms.ml-1 for 24 h to induce interleukin-2 (IL-2) and for 40 h to induce NK cytotoxic factor (NKCF). Radioactivities of [3H]TdR uptaken by CTLL-2 cell line and YAC-1 cell line were used to measure the IL-2 and NKCF activities, respectively. PAP-I enhanced the production of IL-2 and NKCF. These results suggest that PAP-I augments the immunological functions in vivo. PMID- 8237402 TI - Pharmacokinetics and relative bioavailability of lomefloxacin preparations in 10 healthy Chinese volunteers. AB - The pharmacokinetics of lomefloxacin tablet and capsule were determined following a single oral dose of 400 mg given to each of 10 Chinese healthy male volunteers in an open, randomized crossover study. Drug concentrations in plasma were assayed by HPLC method. The peak levels in plasma averaged 6.0 +/- 1.3 and 5.9 +/ 1.0 micrograms.ml-1 at 1.3 +/- 0.4 and 1.2 +/- 0.4 h, and the areas under the drug concentration curves were 43 +/- 15 and 44 +/- 13 h.micrograms.ml-1 for lomefloxacin tablet and capsule, respectively. The concentration-time courses after medication conformed to a 1-compartment open model with a first order absorption. Pharmacokinetic parameters after tablet did not differ significantly from the corresponding values after capsule. The bioavailability of tablet was comparable to that of capsule. PMID- 8237403 TI - [Triggered activities in cat heart induced by combined administration of ouabain and calcium gluconate]. AB - The pharmacological effects of i.v. a mixture of ouabain (Oua) (50 micrograms.kg 1) and calcium gluconate (Ca2+) (100 mg.kg-1) on the electric activities of the cat hearts (n = 19) were studied by using a contact electrode and a contact electrode catheter to record both epicardial and endocardial monophasic action potentials (MAP). The results showed that the 2 drugs together reduced the MAP amplitude, decreased Vmax, and lengthened the MAP duration. Ouabain induced triggered activities, eg, early afterdepolarization, early afterhyperpolarization, delayed afterdepolarization including oscillatory afterpotentials, and triggered arrhythmias, which were enhanced by calcium gluconate. Endocardium is more sensitive than epicardium to ouabain. PMID- 8237404 TI - [Effects of 6,7-dimethoxycoumarin on left ventricular function and hemodynamics in anesthetized cats]. AB - Scoparone (6,7-dimethoxycoumarin 1.25, 2.5, 5 mg.kg-1) influenced the cardiohemodynamic functions in anesthetized cats. After i.v. injection the LVP dP/dt loops and blood pressure were decreased. Both the contractile and relaxant functions of the left ventricle were inhibited. These were demonstrated by the reduction of dP/dtmax, Vmax, Vce-dp40, LVSP, prolongation of LVET, diminution of dP/dtmax, and extension of the constant time of isometric diastolic pressure decay (T value). The preload of the left ventricle, LVEDF, was raised and the afterload (MAP, TPB) was decreased: The systolic volume, systolic index, cardiac output, and cardiac index were decreased slightly. The parameters TTI and TTI x HR, which reflect the myocardial oxygen consumption, were reduced. Furthermore, the heart rate was decreased at 5 mg.kg-1. The results indicate that scoparone could suppress the myocardial activity. PMID- 8237405 TI - [Pharmacokinetics and pharmacodynamics of two morphologically homogenous forms of famotidine per os in Chinese healthy volunteers]. AB - The lg C of famotidine (Fam) A and B forms in plasma vs time curve following a single oral dose of 40 mg showed an one-compartment open model in 5 healthy volunteers. The T1/2Ke of Fam A and B forms = 3.06 and 3.48 h, Tmax = 2.96 and 2.68 h, The Cmax = 115 and 145 ng.ml-1, AUC = 811 and 1190 h.ng.ml-1, respectively. No significant difference was found in the pharmacokinetic and pharmacodynamic properties between Fam A and B forms. The mathematical model describing the whole course of blood concentration of Fam A and B forms in relation to inhibiting effects on gastric acid were: E (A) = 100.C2.04/(C2.04 + 15.0(2.04)) and E (B) = 100.C1.67/(C1.67 + 14.0(1.67)). Predication of blood drug concentration from pharmacodynamics or vice versa became possible using the mathematical equations. PMID- 8237406 TI - [Effects of ranitidine on ventricular refractory period and fibrillation threshold of normal and ischemic ventricle in rabbits]. AB - Effects of ranitidine (Ran) on ventricular refractory period and fibrillation threshold of normal and ischemic ventricle were investigated in pentobarbitone anesthetized rabbits. In saline-treated rabbits, relative refractory period (RRP) and effective refractory period (ERP) were shortened from 132 +/- 24 to 126 +/- 25 ms (n = 10, P < 0.05) and from 118 +/- 22 to 108 +/- 27 ms (n = 10, P < 0.05) in boundary zone of ventricular ischemic regions, respectively. The ventricular fibrillation threshold (VFT) was decreased from 5.5 +/- 1.4 to 2.8 +/- 1.1 V (n = 10, P < 0.01) after coronary artery ligation (CAL). Ran (25 mg.kg-1, i.v.) not only prevented these changes, but also lengthened RRP from 123 +/- 14 to 136 +/- 10 ms (P < 0.05) and ERP from 111 +/- 16 to 126 +/- 12 ms (P < 0.05), and increased VFT from 4.7 +/- 0.8 to 7.4 +/- 1.8 V (P < 0.01) in normal ventricle. The results suggest that Ran may have anti-arrhythmic effects in rabbits. PMID- 8237407 TI - [Effect of quercetin on chemiluminescence of human platelets induced by arachidonic acid]. AB - Arachidonic acid (AA)-induced platelet chemiluminescence (CL) was measured with a lumiphotometer. Quercetin remarkably inhibited the CL, the IC50 of quercetin was 3 mumol.L-1. When quercetin plus aspirin, which inhibits only cyclooxygenase, was added, the inhibitory rate of platelet-CL obviously increased (P < 0.01). On the other hand, the quercetin had a scavenging effect on superoxide anion radical using alkaline sodium dithionite solution generation. The IC50 was 20.9 mumol.L 1. In addition, superoxide dismutase of 0.1 mg.ml-1 inhibited the platelet-CL by 97.8%, while mannitol, a hydroxyl radical scavenger, only by 43.3% at a concentration of 80 mg.ml-1. These results suggest that the mechanism of the inhibiting AA-induced platelet-CL by quercetin was associated with scavenging the superoxide anion radical directly and with inhibiting the cyclooxygenase. PMID- 8237408 TI - [Effects of methylflavonolamine on TXA2 and PGI2 plasma levels in rabbits with myocardial infarction]. AB - Effects of methylflavonolamine (MFA) on TXA2 and PGI2 plasma levels in rabbits of myocardial infarction were investigated. It was found that TXB2 levels were inhibited by MFA 2.5 or 5.0 mg.kg-1 i.v. while 6-keto-PGF1 alpha levels were only slightly inhibited after repeated doses of 5.0 mg.kg-1 i.v. within 6 h. This result indicates that MFA exhibit stronger inhibition on cyclo-oxygenase in platelets than in endothelial cells. In addition, the myocardial injury and the size of myocardial infarction were much less in the MFA treated groups than in the control. PMID- 8237409 TI - [Antioxidative action of soyasaponin monomers I, A1, A2, and their inhibitory effects on action potential and beating of myocardiocytes in culture]. AB - Ventricular myocardiocytes from newborn Wistar rats were cultured. Soyasaponin monomers I, A1, A2 2 micrograms.ml-1 added to the culture medium, like nimodipine 1.25 microgram.ml-1, decreased the number of spontaneously beating clusters and the action potential parameters of myocardiocytes, which restored after washing out and were reversed by Ca2+ 80 micrograms.ml-1 or epinephrine 10 micrograms.ml 1, indicating the Ca2+ channel blockade action of the saponin monomers. Adding xanthine 0.42 mmol.L-1 and xanthine oxidase 5.4 nmol.L-1 into culture medium led to the increase of free radical content of the myocardiocytes and the decrease of action potential parameters, which were reversed by I 2 micrograms.ml-1, indicating the antioxidative action of saponin monomer I. PMID- 8237410 TI - [Immunopotentiating effects of polysaccharides isolated from Medicago sativa L]. AB - In vitro experiments showed that 250-500 micrograms.ml-1 of the polysaccharides isolated from Medicago sativa L (MSP) increased mouse lymphocyte uptake of [3H]thymidine (TdR) induced by PHA-p (0.15%), Con A (5 micrograms.ml-1), LPS (20 micrograms.ml-1), and PWM (2 micrograms.ml-1) at maximum by 5.13 +/- 0.81, 0.75 +/- 0.08, 4.52 +/- 0.36, and 0.86 +/- 0.24 folds, respectively. After MSP ip for 5 d, the average count of mouse splenic lymphocytes in 125 mg.kg-1 x d-1 group increased from 1.4 +/- 0.4 (x 10(7))/mouse to 3.6 +/- 0.5 (x 10(7))/mouse; in 250 mg.kg-1 x d-1 dose group increased to 13.5 +/- 0.3 (x 10(7)). MSP 125 mg.kg-1 x d 1 or 250 mg.kg-1 x d-1 ip for 5 d partially or completely antagonized the effect of cyclophosphamide (200 mg.kg-1 x d-1 x 1 d) on lymphocyte decrease, the average number of lymphocytes/mouse recovered from 1.04 +/- 0.23 (x 10(7)) to 2.1 +/- 0.4 (x 10(7)) and 2.9 +/- 0.6 (x 10(7)), respectively. In addition, the lymphocyte reactivity to Con A in MSP ip 125 mg.kg-1 x d-1 x 5 d group enhanced by 60%, in 250 mg.kg-1 x d-1 x 5 d group by 156%. The abilities of PWM-induced antibody production were elevated 51% and 78%, respectively. PMID- 8237411 TI - [Protective effects of nitrendipine against myocardial infarction in rabbits]. AB - In rabbits the left circumflex coronary artery was ligated for 24 h. Nitrendipine (Nit) 10 mg.kg-1 ig were given 1 h before ligation and 1, 4, and 7 h after ligation. Nit decreased myocardial infarctive size (MIS) from 25.2 +/- 1.7% to 15.5 +/- 1.6% (P < 0.01), lowered serum creatine kinase (CK) and lactate dehydrogenase isoenzyme 1 (LDH1) from 381 +/- 69 IU.L-1 and 42 +/- 6% to 281 +/- 49 IU.L-1 (P < 0.01) and 31 +/- 8% (P < 0.05), diminished injury of myocyte ultrastructure, especially mitochondria and myofibrillae, in ischemic and infarctive zone. Nit exerted no significant effect on plasma level of norepinephrine (NE). The results indicate that Nit has protective effects against the acute myocardial infarction. PMID- 8237412 TI - [Purification and anticancer activity of cytotoxin-14 from venom of Naja naja atra]. AB - Ion-chromatography of lyophilized cobra (Naja naja atra) venom on SP-Sephadex C 50 yielded 14 fractions, of which 7, 8, 9, and 10 possessed neurotoxic actions and 11, 12, 13, and 14 had cytotoxic activities. Chromatography of fraction 14 on SP-Sephadex C-25 gave cytotoxin-14. It was homogeneous on polyacrylamide gel electrophoresis with m = 7448 Da. It was a typical cytotoxin without phospholipase A2 activity and had a selective cytotoxic action on human cancer cell lines. PMID- 8237413 TI - [Protective effect on ischemic depletion of nucleotide phosphates in heart, brain, and kidney by cyclophosphamide]. AB - The levels of ATP, ADP, and AMP in heart, brain, and kidney suffering from 10-min ischemia after decapitation in rats were determined by a modified reverse-phase HPLC set with uv detection. The ischemic depletion of ATP was alleviated and the total amount of high energy phosphates was markedly reduced by the treatment of po cyclophosphamide 20 and 100 mg.kg-1 x 3 d. The protective effect on depleting the total amount of high energy phosphates which was better preserved than ATP in ischemic organs by cyclophosphamide was evidenced in a dose-related manner. Cyclophosphamide induced leukopenia in circulating blood. Two reasons for the anti-arrhythmic effect of cyclophosphamide are suggested: 1) the depletion of leukocyte reduced the plugging effect of neutrophil in myocardial capillaries; 2) blocking the KATP channel by elevating ATP level in myocardium. PMID- 8237414 TI - Effect of ethanol on hippocampal neurons depends on their behavioural specialization. AB - Acute effect of ethanol on hippocampal neurons was studied during food acquisition behaviour in seven rabbits. The rabbits were taught to acquire food from a feeder by pressing a pedal on the same side of the cage. The behaviourally specialized units (L units related to newly learned behaviour and M units related to behaviour formed before learning, e.g. certain movements) were comparable with the 'place' (projectional pyramidal and granular cells) and 'displace' (non pyramidal interneurons) units of the current classification. The same direction of ethanol effects was found as for the limbic cortex; the number of certain kinds of L units decreased and that of M units increased but there was no significant change in the relative number of L and M units as a whole. The background frequency of L units decreased, but the frequency within activations increased. The results confirm our earlier findings on the most marked depressive effect of ethanol on L units and show that it is the behavioural specialization, not the morphological unit type, which is a major determinant of the ethanol influence. PMID- 8237415 TI - Low doses of ethanol may induce anti-nociceptive effects via an oxytocinergic mechanism. PMID- 8237416 TI - Acute renal denervation influences the tubuloglomerular feedback mechanism. PMID- 8237417 TI - Atrial natriuretic factor increases peritoneal fluid formation in the pig. PMID- 8237418 TI - The role of sarcoplasmic reticulum in endothelium-dependent and endothelium independent rhythmic contractions in the rabbit mesenteric artery. AB - Upon stimulation with phenylephrine, the rabbit mesenteric artery displays endothelium-dependent and endothelium-independent rhythmic contractions in the absence and the presence of ryanodine, respectively. For examination of the involvement of the sarcoplasmic reticulum (SR) in these two types of rhythmic contractions, the mesenteric ring was suspended in an organ chamber for isometric tension recordings. Phenylephrine induced endothelium-dependent rhythmic contractions (EDRC), which were converted to endothelium-independent rhythmic contractions (EIRC) by the subsequent addition of ryanodine. Cyclopiazonic acid (CPA) also induced EIRC in the artery contracted with phenylephrine. The nifedipine-treated artery displayed neither EDRC upon phenylephrine stimulation nor EIRC by the addition of ryanodine or CPA: however, these agents relaxed the arteries. Phenylephrine induced EDRC in the artery treated with the K+ channel antagonist sparteine, but these rhythmic contractions were converted to a sustained contraction by ryanodine and CPA without producing relaxation of the artery. Ryanodine and CPA inhibited both phenylephrine-induced Ca2+ release from the SR and Ca2+ sequestration, without affecting Ca2+ influx across the plasmalemma, evaluated by monitoring agonist-induced contractions. These findings indicate that: (1) the EDRC may be attributed to Ca2+ release from the SR, which may be charged by Ca2+ influx via the voltage-dependent Ca2+ channel; and (2) the EIRC may arise from functional impairment of the SR and by the subsequent increase in the K+ efflux, presumably via the Ca(2+)-activated K+ channel. PMID- 8237419 TI - Evidence for an adrenergic innervation of the adrenal cortical blood vessels in rats. AB - The aim of this study was to investigate the blood flow in the adrenal cortex of the rat. Relative changes in the adrenal cortical blood flow were continuously measured by Laser Doppler flowmetry in 33 chloralose-anaesthetized artificially ventilated rats during electrical stimulation (1 ms, 5 V) of the left great splanchnic nerve (LGSN), which conveys both pre- and post-ganglionic nerve fibres to the adrenal gland. Laser Doppler flux (LDF) was decreased and regional resistance (RR) was increased by augmenting nerve stimulation at increasingly higher frequencies (2, 4, 8, 20 and 40 Hz). The decrease in LDF, when compared to pre-drug stimulations at 4 Hz was partially or totally inhibited by the adrenergic blocking agents trimethaphan (TRIM), guanethidine (GUA) and alpha 1 blockade with prazosin (PRAZ). Furthermore, both the decrease in LDF and the increase in RR were either completely or partially blocked by stimulation at 40 Hz after TRIM-treatment and GUA-treatment. It is concluded that the adrenal cortex in the rat is innervated by post-ganglionic adrenergic nerve fibres, which are involved in the regulation of blood flow in the adrenal cortex. PMID- 8237420 TI - Effects of adrenergic stimulants on the splenic diameter, haemoglobin content and haematocrit in anaesthetized dogs: determination of the adrenoceptor subtype responsible for changes in the splenic diameter. AB - Changes in splenic diameter measured by sonomicrometry in response to various adrenergic stimulants were estimated together with simultaneously measured arterial haemoglobin content (HGB) and haematocrit (HCT) in anaesthetized dogs. Splenic diameter decreased following intravenous injections (i.v.) of adrenaline, noradrenaline and phenylephrine and splenic nerve stimulation associated with increases in arterial HGB and HCT, which were significantly attenuated by prazosin i.v. After prazosin i.v., adrenaline i.v. increased splenic diameter significantly, but noradrenaline i.v. did not. Isoprenaline i.v. increased splenic diameter transiently, followed by a decrease that was abolished by prazosin i.v. During occlusion of splenic arteries and veins, adrenaline i.v. and phenylephrine i.v. did not cause any change in arterial HGB and HCT. Injection to splenic artery (i.a.) of phenylephrine induced a significant decrease in splenic diameter that was attenuated by prazosin i.a. but not by yohimbine i.a. Clonidine i.a. did not change splenic diameter. The present results indicate that splenic contraction, which is mediated through alpha 1-adrenoceptor activation, causes a significant increase in arterial HGB and HCT. PMID- 8237421 TI - Breakdown of membrane choline-phospholipids induced by endogenous and exogenous muscarinic agonist is potentiated by VIP in rat submandibular gland. AB - The outflow of tritium from rat submandibular gland fragments, pre-labelled with [3H]choline, following electrical or pharmacological stimulation was studied. Electrical stimulation of the tissue increased the outflow of tritium in a frequency dependent manner. Atropine treatment decreased the electrically-induced release, indicating that the outflow did not reflect acetylcholine from nerve endings, but was largely brought about by postsynaptic receptors. In agreement with this hypothesis, treatment with noradrenaline or carbachol induced a dose dependent increase in tritium outflow from the gland fragments which could be blocked by prazosin or atropine, respectively. Moreover, analysis of the tissue associated tritium revealed an incorporation primarily in the lipid fraction of the tissue (almost 80%), of which about 90% was in phosphatidylcholine, indicating that this was the source of the tritium outflow. Pre-incubation with vasoactive intestinal polypeptide (VIP), which coexists with acetylcholine in the parasympathetic neurons innervating the submandibular gland, increased the carbachol-induced tritium overflow significantly. The effect of VIP could be imitated by the adenylyl cyclase stimulator forskolin, which increased the carbachol-stimulated tritium efflux in a dose dependent manner. Taken together, our results suggests that muscarinic- and alpha 1-receptor agonists may activate a phospholipase coupled to phosphatidylcholine hydrolysis in the rat submandibular gland. Endogenous acetylcholine released from parasympathetic nerve endings appear to activate this mechanism. Furthermore, VIP treatment, and the concomitant cAMP-accumulation, potentiates the acetylcholine induced phosphatidylcholine hydrolysis, demonstrating a new type of interaction between the classical transmitter acetylcholine and the co-stored neuropeptide VIP. PMID- 8237422 TI - Potentiation of serotonin-induced contractility of gastric fundus strips in lactating rats. AB - Gastric fundus strips isolated from lactating, non-lactating and pregnant rats were used to obtain agonist-induced contractions in vitro and the mechanism of alteration in gastric contractility to serotonin during lactation was investigated. The gastric contractile responses to 5-HT expressed as area per unit mass of muscle (cm2 g-1 muscle) were 60-65% greater in lactating rats compared with non-lactating rats. However, responses evoked by acetylcholine (ACh) or histamine were not different. The EC50 values for 5-HT were not different in either group indicating that there was no alteration in the 5-HT affinity for its receptors in lactating rats. In pregnant rats, 5-HT-induced gastric responses were significantly (P < 0.05) lower than those of non-lactating rats. Pretreatment of non-lactating rats with haloperidol (which increases plasma prolactin levels) enhanced the gastric contractile response to 5-HT (P < 0.05). On the other hand bromocriptine administration (which lowers plasma prolactin levels) in lactating or immature rats, decreased the contractile response to 5-HT significantly, while bath application of bromocriptine (0.1 microM) had no effect. Incubation of fundus strips in physiological solution containing prolactin (10 micrograms ml-1) for 24 h decreased the 5-HT evoked contractions, but not the ACh evoked responses. The results in this study indicate that prolactin modulates the intrinsic contractile activity of the gastric smooth muscles to 5-HT. PMID- 8237423 TI - Measurements of duodenal wall motility, mucosal fluid transport and alkaline secretion. Description and evaluation of a methodological approach in the anaesthetized cat. AB - This paper describes and evaluates a method for simultaneous recordings of duodenal motility and mucosal secretions in chloralose-anaesthetized cats. Biliary and pancreatic secretions were diverted by separate cannulation of each duct. A 2 cm segment of the proximal duodenum was isolated between two luminally situated balloons (the proximal balloon being positioned immediately distal to the pylorus). Each balloon was connected to a barostatic device maintaining a constant pressure within the balloon independent of the degree of contraction. Motor activity in the duodenal wall was recorded as changes in balloon volume. The duodenal segment was perfused at a constant rate with isotonic saline containing [14C]PEG 4000 as a non-absorbable marker. The recovered perfusate was sampled in 5 min fractions and analysed with regard to alkalinity (back titration) and concentration of marker (liquid scintillation). Net alkalinization and net fluid transport was calculated with conventional equations. The sensitivity of the analytical procedures was tested bench-side. The technique was evaluated in vivo during basal conditions and during acid exposure of the mucosa. Exposing the duodenal segment to 30 mM HCl induced a marked change in the duodenal functional state with the occurrence of a polarized duodenal motility pattern, net fluid secretion and an increased alkaline secretion. PMID- 8237424 TI - Cardiovascular changes during an isometric contraction combined to a cold pressor test. AB - Fifteen normotensive male subjects (age, 27.9 +/- 1.3 years) performed isometric contractions (handgrip) at 15, 30 and 50% maximal voluntary contraction respectively for 7, 3 and 1 min. The contractions were sustained with or without a cold pressor test of the same duration (immersion of one foot in water of 5.2 +/- 0.1 degrees C). At rest, under the influence of a cold pressor test for 3 min, the heart rate increased from 74.3 +/- 2.2 to 83.3 +/- 2.1 beats min-1 (P < 0.001) in less than 10 s, and returned to the control value in 3 min; on the contrary, at the end of 3 min of cold pressor test, the systolic blood pressure was still above the control value (135.5 +/- 4.2 and 121.0 +/- 3.2 mmHg, P < 0.001). During the contractions, there was no significant difference between heart rate and systolic blood pressure values obtained with or without cold pressor test. Similarly, there was no significant change in the concentrations of plasma catecholamines. This absence of cardiovascular effect of cold pressor test associated with isometric contraction (for three relative force levels) is probably due to the increasing importance of sympathetic cardiac activation produced by the contraction with respect to that produced by the cold pressor test, the absence of supplementary changes in total peripheral resistance and a partial reciprocal inhibition of pain produced by the simultaneous performance of these two tests. PMID- 8237425 TI - Renal effects of urodilatin and atrial natriuretic peptide in volume expanded conscious dogs. AB - The renal effects of urodilatin and atrial natriuretic peptide (ANP) were examined in conscious dogs during acute volume expansion maintained through independent infusions of water and NaCl. Peptide was infused in a step-up fashion, in 40-min periods at rates of 2.5, 12.5 and 50.0 ng kg-1 min-1 (ANP, urodilatin) or 0.5, 2.5 and 10.0 ng kg-1 min-1 (second series of urodilatin). ANP immunoreactivity (ANPir) in plasma was measured with an antibody showing 100% cross-reactivity with urodilatin. At 50 ng kg-1 min-1, ANP increased plasma ANPir some 10-fold (64 +/- 6 pg ml-1 to 728 +/- 82 pg ml-1) while urodilatin elicited a 25-fold increase in ANPir (78 +/- 13 pg ml-1 to 1645 +/- 204 pg ml-1). ANP doubled sodium excretion (61 +/- 12 to 124 +/- 30 mumol min-1, P < 0.05) while urodilatin increased sodium excretion to 269 +/- 45 mumol min-1. Both peptides approximately doubled urine flow. Urodilatin at 10 ng kg-1 min-1 increased sodium excretion from 57 +/- 17 mumol min-1 to 106 +/- 25 mumol min-1, i.e. similar to the response ANP elicited at 50 ng kg-1 min-1. In the time control series a significant fall in sodium excretion was observed, despite the continuous replacement of sodium and water. The results demonstrate that in identical, relatively high doses, the changes in sodium excretion, diuresis, heart rate (HR) and arterial pressures elicited by urodilatin are more pronounced than those of ANP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237426 TI - Glycogen breakdown and lactate accumulation during high-intensity cycling. AB - High-intensity exercise results in a large breakdown of glycogen. The glycogen lost may reappear as hexose phosphates, lactate, or it may be fully oxidized. Part of the lactate produced may be transferred from muscle to blood. There is, however, incomplete information on the relative importance of each endpoint of glycogen breakdown during high intensity exercise. Therefore, 16 healthy men cycled for between 30 s and 3 min until exhaustion. Muscle biopsies were taken from m. vastus lateralis before and immediately after exercise and analysed for glycogen, glucose, glucose-6-phosphate and lactate. In addition the blood lactate concentration was measured at exhaustion, and the O2 uptake was measured throughout the exercise for calculation of glycogen oxidation. The muscle glycogen concentration fell by 17-24 mmol kg-1 wet wt muscle, the muscle glucose and G-6-P concentrations rose by 1 and 4 mmol kg-1 respectively, and the muscle lactate concentration rose by 20-30 mmol kg-1. The blood lactate concentration at exhaustion was 4-9 mmol l-1 above pre-exercise value. Consequently, 60% of the glycogen lost reappeared as lactate within the working muscle, another 20-25% was found as other glycolytic intermediates, 4-13% of the glycogen loss could be accounted for by oxidation. Lactate released to blood could account for approximately 10% of all lactate produced. Therefore, when large muscles are heavily engaged, as during high intensity cycling, most of the glycogen broken down appears as lactate within the working muscle. PMID- 8237427 TI - The effects of oestrogen and progesterone on insulin sensitivity in female rats. AB - The effects of 17-beta-oestradiol (E2) and progesterone (P) on insulin sensitivity were determined in oophorectomized (OVX) rats by the euglycaemic hyperinsulinaemic clamp technique combined with measurements of insulin stimulated 2-deoxy-D-glucose (2-DOG) transport and glycogen synthesis in white and red parts of the gastrocnemius, the extensor digitorum longus and soleus muscles as well as in the liver (only glycogen synthesis). OVX was followed by insulin resistance in the clamp measurements. This was paralleled by a decreased insulin-stimulated content of 2-DOG in muscles, an index of glucose transport. Glycogen synthesis in muscle was also decreased, although to less extent. E2, alone or in combination with P, restored this to values of intact controls, while P alone was followed by insulin resistance. Liver glycogen synthesis was also decreased by OVX but this required combination of E2 and P to be fully restored. It was concluded that particularly E2 plays an important role in the maintenance of normal insulin sensitivity while P alone seems to be followed by insulin resistance, both effects apparently mainly by regulation of glucose uptake in muscle. E2 + P may be of importance for maintenance of normal glycogen synthesis in the liver. PMID- 8237428 TI - Altering the blood-brain barrier in the rat by intracarotid infusion of polycations: a comparison between protamine, poly-L-lysine and poly-L-arginine. AB - To evaluate the role of surface charge for the blood-brain barrier permeability, the albumin content was determined in the cerebrospinal fluid and in the brain 1 h after intracarotid infusion of protamine sulphate, a natural polycationic protein with a high content of arginine (mol. wt 4000-4400), poly-L-arginine (mol. wt 11,600) or poly-L-lysine (mol. wt 10,200). Five milligrams (4 x 10(-4) mmol) poly-L-arginine increased the albumin content in the brain 15 times more than 5 mg (5 x 10(-4) mmol) poly-L-lysine (P < 0.001) and 3.5 times more than 5 mg (1 x 10(-3) mmol) protamine (P < 0.001); the difference between protamine and poly-L-lysine was also significant (P < 0.05). After 0.5 mg (4 x 10(-4) mmol) poly-L-arginine the albumin extravasation was still higher than after 5 mg protamine (P < 0.01) and 5 mg poly-L-lysine (P < 0.001). Cisternal albumin increased from control values 0.08 mg ml-1 to 0.30, 0.46 and 1.21 mg ml-1 in rats given 5 mg poly-L-lysine, protamine and poly-L-arginine, respectively (P < 0.01 for difference between arginine and the other two substances). The higher mol. wt and positive charge of poly-L-arginine may at least in part explain the more pronounced albumin leakage after arginine than after protamine. However, the difference between poly-L-arginine and poly-L-lysine suggests that other factors, possibly related to the guanidino groups, contribute to the blood-brain barrier opening by poly-L-arginine. PMID- 8237429 TI - [Psychiatry and history]. PMID- 8237430 TI - [Perspectives in psychiatric research in Latin America for the 21st century]. AB - Psychiatric research in Latin America lacks in originality and methodological rigor, and the human and economic resources are extremely limited. However, there are groups in several countries that have used their initiative and resources with a high level of efficiency, particularly in the psychosocial area. Epidemiological investigation in Latin America is exemplary and can, indeed, be one of the most significant contributions of our psychiatry. Assisted by social scientists, psychiatric researchers can study the most pressing problems in the region (alcoholism, substance abuse, violence, problems with children, adolescents and the elderly), as well as traditional areas of clinical investigation such as neuroses and psychoses. Biomedical investigation will continue in selected centers with a clear understanding of priorities and costs. Psychiatric research in Latin America requires realism and gradulism in its planning, multidisciplinary and intercountry collaboration, adequate publication of its findings, and academically trained personnel. Some countries have accumulated expertise in specialized areas and that, together with the contribution of Latin American researchers in other continents, should be positively utilized. PMID- 8237431 TI - [A neurobiological model of mind]. AB - An essential difference is established between the dorsal, and ventral regions of the cerebral cortex: While dorsal cortex--linked to archicortex or hippocampal allocortex--elaborates acts, and objects used in, or by, acts, ventral cortex- linked to paleocortex or olfactory allocortex, builds up preventive values favoring the inhibition of acts whenever the use of objects may become dangerous. Such a distinction between the dorsal and ventral regions of the cerebral cortex is founded mainly on disinhibition syndromes appearing in patients with ventral lesions--besides, the said distinction can explain some disinhibition demonstrations included in the schizophrenic clinical pictures. PMID- 8237432 TI - [Molecular chromosome mapping in psychiatric diseases]. AB - This paper describes the procedure to obtain some DNA probes likely to be good candidate genes for mapping major psychiatric disorders. The advantages in using this mapping strategy are discussed as well. PMID- 8237433 TI - [Ego strength evaluated through the Bender test]. AB - This article is aimed at observing the ego strength among a psychosomatic patients sample quantitatively. The Bender-Gestalt test (as per Pascal & Suttell's method) was administered to the above mentioned sample as well as a psychotic patients sample, and a control group. No differences were observed between the psychosomatic patients group, and the psychotic patients group performance. However, differences were observed when comparing with the control group performance. PMID- 8237434 TI - [Anxiety and depression levels in rape offenders]. AB - Whether the active subject of rape possesses a particular personality trait is a much debated matter. It is an accepted fact that a small amount of rapists fall into the sadistic diagnostic category. Other psychopatologic traits likely to identify a rapist are, however, unknown. The present study was carried over at a jail near the city of Xalapa (Veracruz) Mexico. Hamilton, IDARE, and Zung scales, in their modified Spanish version were applied to a sample of imprisoned adult rapists. Compared to their control group (men serving sentences for other felonies), rapists scored higher in anxiety, and depression scales. In the light of these results, it is suggested that some rapists are likely to be anxious depressive subjects that, after having "acquired" a sociopathic repertoire, offend other people in conditions of indefensiveness, contrary to depressive subjects who aggress themselves. PMID- 8237435 TI - [Mini-Mult 82: review and standardization of short version of the MMPI questionnaire]. AB - The correction norms for the Mini-Mult 82 (71 items from Kincannon's MMPI short form, plus 11 representative items from 5(Mf) Scale from the same test--developed by the authors--) are presented. A 1,343 normal, over 15-year-old Costarrican subjects sample was tested. Correction norms were obtained successfully. More reliable scores were obtained by using the Mini-Mult 82 as an independent test (i.e. without MMPI's score predictions). Statistical differences in F, 1(Hs), 2(D), 3(Hy), 4(Pd), 5(Mf) and 8(Sc) scales were found when gender was controlled. "T" scores were computed for raw scores on each scale. Additional research is suggested to improve the test's reliability and validity characteristics. PMID- 8237436 TI - [Depression in pregnancy and puerperium: study of risk factors]. AB - This study was aimed at determining risk factors associated to depression in pregnancy and/or the puerperium. By means of both observer-rated, and self-rated scales a 108-expectant mother sample was assessed by the co-investigators, interviewed by the principal investigator later on. The Psychiatric Assessment Schedule was used to obtain an RDC diagnosis. The same procedure was repeated 2 to 3 months after delivery. A greater exposure to life events, prior consultation for emotional problems as well as economic difficulties arising of late were all associated to depression in pregnancy and the puerperium. Getting separated from significant persons in a woman's life, and/or difficult relationships with either husband or mother have been strongly associated to depression at both periods. As regards puerperium depression, several associations directly related to childbirth (having had a low-weight baby) or the postnatal period (lactation problems) were detected. Anxiety and depressive symptoms during pregnancy have been also clearly linked to postpartum depression. The predictive value of such risk factors is discussed. It is suggested that questions on these factors should be included into ordinary health assessments of both pregnant and puerperal women. PMID- 8237437 TI - [The first case of melancholia in Rio de la Plata]. AB - Alonso Cabrera's is the first recorded case of melancholy in the history of the River Plate area so far. It is the author's contention that melancholy, as it appears in Western countries nowadays, could not have appeared prior to the Renaissance period: Indeed, it was necessary that Europeans were in a position to achieve a sufficient point of personal differentiation so that they could feel responsible for their acts by means of guilt--hence falling into melancholy. PMID- 8237438 TI - [More about panic disorders]. PMID- 8237439 TI - [Borderline personality disorders (ACTA 1992, 38 (4) 323)]. PMID- 8237440 TI - [Education and mental health]. PMID- 8237441 TI - [Semantic information. Internal language. Thinking]. AB - Semantic information has reached an objective condition after a lengthy history of semantic inquiries that instrumental neurophysiological devices--such as event related potentials, electroencephalographic spectral analysis, regional brain circulation, PET scan, deep brain electrodes, and other--have made easier. In turn, internal language, as screened according to Vigotsky's perspective, is considered a product of semantic information circulation understood as neurosemae interconnection. Finally, in normal adults, thinking processes are assumed to be made up by both sensoperceptive information (proprioceptive information included) and semantic information. Thus, an "extraverbal thinking" can be distinguished, whose activity is hardly describable in healthy adults but should be considered as a condition of non-educated deaf persons, and a "verbal thinking", or internal language, made up by semantic information. PMID- 8237442 TI - [Biological markers in psychiatry]. AB - Recent advances on the biological markers in Psychiatry are reviewed in this paper. For systematization's sake, the available information has been divided into five major areas: Neuroimage, neurophysiology, psychoendocrinology, neurochemistry and genetics, and molecular biology. Data generated by research in this field have extended our knowledge on mind functioning although their clinical applications are still limited for the time being. PMID- 8237443 TI - [Integration in psychotherapy]. AB - The dialectic relationship among trends aimed at either integration or disrupture in the field of psychotherapies over time is discussed. Theoretical, clinical, and social issues sustaining either a tendency toward an integration of the different psychotherapeutic orientations, or their counterparts whenever the different disrupture processes took place in times past are dealt with. PMID- 8237444 TI - [Periodicities in the longitudinal evaluation of affective status]. AB - Both circadian and infradian rythms in Affective State (AS) are reported in 12 subjects (10 with Affective Disorders, 2 without Mental Disorder). The segmented visual scale ESTA-III was used, which yields scores for two dimensions: Mood and Drive. Self-assessment of AS was carried out every 8 hours for about a month. The statistical procedure is underlined. Theoretical implications of findings are discussed. PMID- 8237445 TI - [Adolescents' values: motivational axes]. AB - The impact of both gender and nationality on the motivational dimensions of human values was assessed on a male/female 140-teenager sample studying at an American high school. Sample was a half-and-half split group between Americans and Brazilians. Sixty values were evaluated: 56 cross-cultural values, and 4 values peculiar to the Brazilian culture. Subgroup differences were checked up at both the motivational type level and the values bipolar dimension's level. Girls placed a stronger emphasis than boys on values related to self-transcendence of selfish concerns, and promotion of other people's welfare. Americans scored higher on achievement values while Brazilians scored higher on hedonistic values. PMID- 8237446 TI - [Adoption: a systemic point of view]. AB - From a systemic viewpoint, communication and structure of adoptive families are analyzed. An isomorphism between the environment interaction with the adoptive family, and the different family subsystems was found out. Within such a conceptualization, the psychological vulnerability of adoptive children is studied. PMID- 8237447 TI - [Beliefs on and attitudes toward mental diseases and the psychiatrist: exploratory study]. AB - Preliminary findings of a research carried out to determine the main beliefs and attitudes general public holds toward three closely related attitudinal objects (namely: Mental illness, the mentally ill, and psychiatrists) are presented. A questionnaire specially designed for this study was applied to a 150-subject sample from the adult general population of Mexico City. Subjects' information level was higher than expected. Their attitudes were also more positive than those foreseen: A majority of subjects recognized both the importance of mental illness, and the advisability of seeking qualified treatment. Nevertheless, information regarding causes, characteristics and course of mental diseases is lacking to a wide extent: In this connection, the need for developing education programs is emphasized. PMID- 8237448 TI - [Alejandro Korn, psychiatrist and philosopher]. AB - Alejandro Korn is a clear example of a man whose life evidences to be firmly coherent between thinking and action. He was a physician, a psychiatrist, and a philosopher, a first-line actor in various decisive events affecting his country, Argentina. PMID- 8237449 TI - Discriminative feature integration by individuals. AB - A tactile pattern on the skin generally comes to be recognised from its function within action. What is perceived tactually and may generate a textural or geometrical sensation is therefore a set of dynamic characteristics that can be extracted by mechanoreceptors and central neural processing from the physics of the interactions of hands, mouth or other parts of the body with common objects and materials. The recognised dynamic (as is true of any sensory modality) may be diagnosable as distinct features that are integrated by the whole system's operation. If so, this transmission of feature information over a channel or in a dimension can be measured as the strength of the influence of that aspect of the stimulating pattern on the observed output, in units of the acuity of response differences for deviations of the pattern from its familiar configuration of features. When applied at the individual level, such multidimensional discrimination analysis provides powerful diagnoses of pattern-recognition processes. PMID- 8237450 TI - Individual differences in sensory preferences for fat in model sweet dairy products. AB - Sensory preferences for fat in foods are influenced both by subject characteristics and by the nature of the food stimulus. Hedonic responses to sugar/fat mixtures depend on strong interactions between perceptions of the two ingredients, including tactile as well as gustatory effects. Fat-containing liquid and solid foods, and dessert-type sugar/fat mixtures produce different hedonic response profiles. Body weight, weight history and the presence of eating disorders are associated with differences in hedonic responsiveness to fat among women. This inter-subject variability in hedonic response profiles mandates the use of novel techniques for data analysis. Conventional hedonic response curves mask between-subject differences. Individual patterns of hedonic responses to sugar/fat mixtures are better represented by frequency distributions of maximum hedonic responses or of individual optimal sugar-to-fat ratios. PMID- 8237451 TI - Effect of size, shape and hardness of particles in suspension on oral texture and palatability. AB - Oral texture and consistency perception and preference depend on the size, shape and hardness of the particles in aqueous suspensions. Assessments of suspensions of model particles of garnet, polyethylene and mica showed that fairly large particles can be palatable if shape and hardness constraints are taken into account. Size differences matter more if particles are hard and sharp than if they are soft and rounded. PMID- 8237452 TI - Oral astringency: a tactile component of flavor. AB - The psychophysical, chemical and physiological bases of the oral sensation of astringency are discussed in an effort to clarify the nature of this enigmatic sensation. It is argued that, despite recent data from animal studies that suggest it is a taste sensation mediated by the chorda tympani nerve, astringency arises primarily from increases in friction between mucosal surfaces. Evidence is discussed that supports the view that oral astringency results primarily from the precipitation of salivary mucoproteins, which impairs the natural lubrication of oral surfaces. The ensuing rise in friction induces sensations of dryness and roughness which, although subtle at first, can grow in magnitude over repeated exposures. These changes in the surface texture of the mouth are an important component of the 'flavor' of common foods and beverages such as fruits, nuts, teas and coffee. PMID- 8237453 TI - Touch and taste in the mouth: presence and character of sapid solutions. AB - Complex dynamic properties of a bolus in the mouth are specified by touch. Cooperation between touch and taste may help to specify the character of the bolus. The role of touch in specifying the presence of the oral bolus, in contrast to it's characteristics, remains a theoretical and experimental challenge. PMID- 8237454 TI - Extracting object properties through haptic exploration. AB - This paper reviews some of our recent research on haptic exploration, perception and recognition of multidimensional objects. We begin by considering the nature of manual exploration in terms of the characteristics of various exploratory procedures (EPs) or stereotypical patterns of hand movements. Next, we explore their consequences for the sequence of EPs selected, for the relative cognitive salience of material versus geometric properties, and for dimensional integration. Finally, we discuss several applications of our research programme to the development of tangible graphics displays for the blind, autonomous and teleoperated haptic robotic systems, and food evaluation in the food industry. PMID- 8237455 TI - Responses of cutaneous mechanoreceptors to the shape of objects applied to the primate fingerpad. AB - The present study is one of a series whose aim is to determine how shape is represented in the activity of cutaneous mechanoreceptive peripheral nerve fibers. Cylindrical bars of varying curvature were indented into the receptive fields of slowly and rapidly adapting mechanoreceptive afferent nerve fibers (SAs and RAs respectively) supplying the fingerpad of the anesthetized monkey. The evoked pattern of nerve impulses in single nerve fibers was recorded electrophysiologically. SAs responded to differences in the curvature, both during the ramp and static phases of the skin indentation. RAs responded only during the ramp phase of the indentation, but their responses were not modulated by differences in curvature. Evidence from the present and previous studies is used to support the following hypotheses: Spatial parameters (such as the 'shapes' or 'widths' of responses rates plotted over the skin surface) of primarily SAs in a spatially distributed population of fibers govern the recognition of the overall object shape as a distribution of curvatures; Intensive parameters (such as the magnitude of discharge rates) of only SAs under static indentations, and both SAs and RAs under stroking, are important for discriminations of small differences in curvatures of objects belonging to the same category of shape. PMID- 8237456 TI - Roughness coding in the somatosensory system. AB - Roughness perception is coded in the somatosensory system by neurons in the type I slowly adapting (SAI) system. When the fingers scan a surface, an isomorphic representation of the surface is encoded in the discharge patterns of SAI afferents. Central neurons in area 3b of primary somatosensory (SI) cortex spatially filter the peripheral image to compute local spatial variation. The outputs from these neurons converge onto neurons in area 1 and onto neurons in secondary somatosensory (SII) cortex which we believe is the critical processing pathway underlying roughness perception. PMID- 8237457 TI - Physicochemical properties accounting for cheese texture. AB - Perception of texture is a complex dynamic series of events which cannot be modelled easily by physicochemical measurements. Results using cheese analogues suggest that moisture and fat are key components in determining texture because they soften the protein matrix and provide weaknesses through which the cheeses fracture. These characteristics are sensed as firmness and work during chewing. Possible models are proposed for further investigation of the tactile determinants of cognitive mechanics involved in cheese texture description. PMID- 8237458 TI - Rheological basis of oral characteristics of fluid and semi-solid foods: a review. AB - Oral viscosity plays an important role in the textural appreciation of fluid and semi-solid foods. Oral assessments of viscosity, thickness, stickiness and sliminess correlate with both small-deformation and large-deformation rheological measurements. Small-deformation measurements of viscosity at an oscillatory frequency of 50 rad.s-1 have the highest correlation with assessed thickness of texture but there is still much speculation as to what forces actually operate in the mouth. PMID- 8237459 TI - Multiple physical patterns in judgements of the creamy texture of milks and creams. AB - The textural sensation generated by dairy creams and milks in the mouth was hypothesized to arise from mechanoreceptor stimulation by the viscosity of these oil-in-water emulsions and features arising from characteristics of the globules of fat. The viscosity, range of sizes of the fat globules and the average distance between globules were varied independently in dairy emulsions and samples rated for creaminess by six assessors. In this brief report, each assessor's performance is tested for multidimensional combinations of discriminations among viscosities, globule-size ranges and interglobule distances. Those individuals whose judgements of creaminess were sensitive to these physical variables appeared to differ greatly in the factors on which they based their judgements. This also depended on whether a high-fat milk or a light cream was used as the standard for comparison. Subject to confirmation of the stability of the discrimination space for an individual using a standard, it is preliminarily concluded that recognition of the physical properties of fat in dairy emulsions is highly idiosyncratic, perhaps indicating that it has to be learned. PMID- 8237460 TI - Differential laminar expression of AMPA receptor genes in the developing rat visual cortex using in situ hybridization histochemistry. Effect of visual deprivation. AB - The postnatal development of laminar pattern of AMPA receptor mRNA subtypes GluR A through to GluR-D (flip variants) in the visual cortex of both normally raised and monocularly deprived rats (one eyelid sutured at the age of 11 days) was studied using in situ hybridization histochemistry and computer-assisted image analysis. The AMPA receptors GluR-A through to GluR-D transcripts exhibit a differential laminar expression pattern in the developing rat visual cortex. At birth the levels of GluR-A transcripts are lower by about 50% in each visual cortical layer as compared to the adult values. In contrast, GluR-B to GluR-D mRNAs are expressed in all cases at higher levels at birth than in the adult brain. Unilateral eyelid closure at postnatal day 11 for several periods of time resulted in both transient and permanent changes in the laminar development of GluR-A through to GluR-C transcripts but hardly affected the GluR-D mRNA subtype. The distinct laminar developmental pattern of AMPA receptor mRNAs in rat visual cortex as well as the differential effects of visual deprivation suggest specific roles of AMPA receptor subtypes during the early postnatal maturation of visual function. PMID- 8237461 TI - Excitatory amino acid response in cultured rat striatal neurons results in a developmentally regulated cGMP formation. AB - Glutamate and its analogues play a central role in excitatory neurotransmission throughout the brain. Their signal in the postsynaptic cells can be transduced by several second messengers. Here we show that in primary cultures of embryonic rat striatum, excitatory amino acid receptor stimulation increases cyclic GMP intracellular concentration and the magnitude of this response depends upon the time in culture. Formation of cyclic GMP appears to be mediated by both N-methyl D-aspartate (NMDA) and non-NMDA type excitatory amino acid receptors, it is blocked by specific excitatory amino acid antagonists and requires extracellular Ca++. The effect mediated via the NMDA receptor is also regulated by extracellular Mg++. These results show that excitatory amino acids make use of cyclic GMP for signal transduction in striatal neurons in vitro. We suggest that cyclic GMP may be an independent second messenger possibly important in the development of a defined population of striatal neurons. PMID- 8237462 TI - Differential development of the stress response in congenital learned helplessness. AB - Early in the development of the hypothalamic-pituitary-adrenal (HPA) axis, the rat undergoes a stress hyporesponsive period of blunted responses to several stressors including cold exposure (CE) and maternal deprivation (MD). We examined the development of the axis by monitoring adrenocorticotropin hormone (ACTH) plasma levels in an animal model of depression and/or anxiety characterized by learned helpless (LH) behavior and a dysfunctional HPA axis in adult life. On postnatal day 7 there was no significant difference in basal plasma ACTH levels between congenital (cLH) and controls, but cLH animals showed a blunted response to CE (P < 0.001). By postnatal day 14 there was a dramatic increase in ACTH response to CE (P < 0.005). On postnatal day 21 baseline ACTH and response to CE were again significantly suppressed in cLH rats. Stress responsiveness to MD was present in all groups and was insignificantly different for all ages of development between groups. These findings suggest that rats with congenital learned helplessness undergo a differential response in the development of the HPA axis in that the axis was hypersensitive at postnatal day 14 and became hyporesponsive beyond day 14, and this may, in part, account for the dysfunctional stress response observed during adulthood. PMID- 8237463 TI - Calcitonin gene-related peptide immunoreactive nerve fibers in the developing salivary glands of the rat. AB - The appearance and distribution of calcitonin gene-related peptide immunoreactive (CGRP-IR) nerve fibers was studied by indirect immunofluorescence microscopy in the developing salivary glands of the rat at daily age intervals from the 15th day in utero (i.u.) until birth, and subsequently on the 2nd, 5th, 7th, 16th, 30th postnatal (p.n.) days. The findings were compared to samples of adult rats. CGRP-IR fibers appeared on day 20 i.u. both in parotid and submandibular glands. In submandibular glands, fibers were first seen surrounding the developing ductal branches, in the mesenchyme and also in association with developing ducts and blood vessels. In the parotid gland CGRP-IR fibers first appeared around ductal branches and blood vessels on day 20 i.u. and on day 21 i.u. CGRP-IR nerve fibers were found in the mesenchyme and around developing ducts. The density of CGRP-IR fibers was highest between the 2nd and 16th postnatal days, after which the amount of CGRP immunoreactivity slowly decreased to the levels of adult glands. The increase in a number of CGRP-IR nerve fibers especially around the ductal and vascular elements in the developing salivary glands indicates that they may play a role in the functional maturation of salivary glands. PMID- 8237464 TI - Action of the diabetogenic drug streptozotocin on glycolytic and glycogenolytic metabolism in adult rat brain cortex and hippocampus. AB - In sporadic Alzheimer's disease (AD), a number of metabolic alterations to the brain have been observed soon after the onset of the initial clinical symptoms. In particular, impairments of glucose utilization and related metabolic pathways are prominent and well-established findings in incipient AD, resembling metabolic abnormalities such as have been found in noninsulin-dependent diabetes mellitus. To mimic these abnormalities, we administered an intracerebroventricular (icv) injection of streptozotocin (STZ) to rats and studied the effects of glucose and glycogen metabolism in the cerebral cortex and hippocampus compared with controls. The enzymatic activities studied dropped significantly by 10-30% in brain cortex (cort.) and hippocampus (hc) 3 and 6 weeks after icv STZ injection: hexokinase (15% 3 weeks cort.; 14% 6 weeks cort.; 12% 3 weeks hc; 28% 6 weeks hc), phosphofructokinase (15%; 15%; 24%; 15%), glyceraldehyde-3-phosphate dehydrogenase (10%; 12%; 30%; 19%), pyruvate kinase (22%; 13%; 22%; 28%), glucose 6-phosphatase (10%; 23%; 14%; 19%) and phosphorylase a (22%; 11%; 30%; 15%). The content of glycogen was significantly higher in STZ-treated rats than in control animals (7% 3 weeks and 15% 6 weeks in cortex). In contrast to the reduced enzymatic activities, we observed no changes in the concentrations of the glycolytic intermediates glucose, glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-diphosphate, pyruvate, lactate and glucose-1-phosphate. These data clearly indicate reduced glycolytic enzyme activity after icv administration of STZ and suggest gluconeogenesis consequent on abnormalities in glucose breakdown. This model may thus be assumed to be a useful tool to investigate pathogenetic factors involved in sporadic dementia of Alzheimer type. PMID- 8237465 TI - Glial fibrillary acidic protein and its encoding mRNA exhibit mosaic expression in a glioblastoma multiform cell line of clonal origin. AB - The expression of two astroglial differentiation markers, vimentin and glial fibrillary acidic protein, was investigated in a previously established human glioma cell line of clonal origin (GL15). Vimentin immunolabelling was homogeneously expressed in all cells. Glial fibrillary acidic protein and its encoding message, investigated by immunocytochemistry and in situ hybridization, showed a mosaic-like expression. Only 30% of the cell population expressed glial fibrillary acidic protein and its mRNA. Western and Northern blots performed for both markers confirmed the presence of both proteins and messages, and their level was correlated with the observed antigenic and molecular probe labelling. The overall antigenic pattern suggests that GL-15 cells do not belong to the O-2A progenitor cell lineage and may arise from a clonal expansion of astrocyte precursors. PMID- 8237466 TI - Ontogenesis of type II spiral ganglion neurons during development: peripherin immunohistochemistry. AB - In this study, we analysed the distribution of the intermediate filament peripherin in the developing cochlea of the rat. At gestational day 16, weak immunolabeling was observed in neuronal somas throughout the spiral ganglion. At gestational day 20, the peripherin labeling increased in intensity throughout the spiral ganglion. At gestational day 20, the peripherin labeling increased in intensity throughout the cochlea but became especially strong in some ganglion neurons of the basal turn. Homogeneous immunolabeling was observed throughout the spiral ganglion of the apical turn. Double immunofluorescence labeling of the prenatal cochlea with peripherin and neurofilament (NF) antibodies revealed colocalization on the same structures. By postnatal day 3, the peripherin labeling intensity had decreased in the majority of spiral ganglion neurons, but remained strong in some cells of the basal turn. Only a few neurons continued to be immunolabeled into adulthood that correspond to Type II spiral ganglion neurons expressing both NF protein and peripherin, two classes of intermediate filament proteins. In the organ of Corti, the first immunolabeling was observed on gestational day 20 as peripheral fibers reaching the receptor cells. Positive fibers were observed below both inner (IHCs) and outer (OHCs) hair cells. At birth and at postnatal day 3, peripherin immunolabeling was still observed below both IHCs and OHCs. By postnatal day 4, peripherin labeling became more dominant in fibers below OHCs, but some immunoreactivity was still present below IHCs. No immunoreactivity was present in the intraganglionic spiral bundle (IGSB) fibers containing the olivary complex efferent fibers before birth. A few days after birth some fibers of the IGSB started to be immunoreactive. PMID- 8237467 TI - Commercially available luminometers and imaging devices for low-light measurements and kits and reagents utilizing bioluminescence or chemiluminescence: survey update I. AB - This survey update was compiled in June 1993 and includes products not covered in the luminometer survey (Jan 1992; Stanley, PE. J Biolumin Chemilumin 1992; 7:77 108 and 7:157-69) and kit and reagent survey (Nov 1992; Stanley, PE. J Biolumin Chemilumin 1993; 8:51-63). Some products are new whilst for others details have only recently become available to the author. Technical details are provided together with company address and contact information. PMID- 8237468 TI - Flow injection determination of glucose, bile acid and ATP using immobilized enzyme reactor and chemiluminescent assay of NAD(P)H. AB - We have developed a chemiluminescent flow injection method for analysis of bile acid, glucose and ATP using the chemiluminescent assay of NADH using 1-methoxy-5 methylphenazinium methyl sulphate (1-MPMS)/isoluminol(IL)/microperoxidase (m-POD) system and immobilized enzyme reactors such as 3 alpha-hydroxysteroid dehydrogenase, glucose-dehydrogenase, hexokinase and glucose-6-phosphate dehydrogenase. The standard curves were obtained in the range of 5-100 pmol for bile acid, 0.5-5.0 nmol for glucose and 10(-7)-10(-5) mol/L for ATP. The coefficient of variation for each assay was not more than 4.1% for bile acid, 2.3% for glucose and 5.3% for ATP, respectively. PMID- 8237469 TI - Effect of prostaglandin E1 on chemiluminescence response and adherence of human polymorphonuclear leukocytes to human cultured endothelial cells of prostaglandin E1 treated polytraumatized patients. AB - We investigated the effect of prostaglandin E1 on human polymorphonuclear leukocytes, in vivo. Polymorphonuclear leukocytes of a prostaglandin E1 and placebo study group were harvested and their function, as production of oxygen derived metabolites and adherence to human cultured endothelial cells, was compared. Additionally, data obtained from polymorphonuclear leukocytes of a prostaglandin E1 and placebo group were compared with data obtained from polymorphonuclear leukocytes from 28 blood donors, who served as a control group. Production of oxygen-derived metabolites by polymorphonuclear leukocytes during contact with endothelial cells was measured by chemiluminescence. Chemiluminescence was significantly (p < 0.01) increased in the placebo group in comparison to the control group decreasing to values of control group after 6 d (post-trauma). Chemiluminescence response was not significantly suppressed in patients treated with prostaglandin E1 in comparison to the placebo group. Adherence of polymorphonuclear leukocytes (placebo group) to endothelial cells was significantly increased (p < 0.01) within the first 6d post-trauma. Following day 6, values were in the same range as values for the control group. Adherence was not significantly suppressed in patients treated with prostaglandin E1 in comparison to the placebo group. In conclusion, prostaglandin E1 at a dose of 20 ng/kg bw/min does not influence production of oxygen-derived metabolites and adherence in polytraumatized patients in comparison to a placebo group. Additionally, production of oxygen-derived metabolites by polymorphonuclear leukocytes in response to endothelial cells is shown and it is evident that endothelial cells might influence production of oxygen derived metabolites by polymorphonuclear leukocytes. PMID- 8237470 TI - Bioluminescence and chemiluminescence literature. Luciferase reporter genes--lux and luc. Part 2. AB - The following references encompass a review of recent literature where the in vivo expression of eukaryotic and/or prokaryotic luciferases provide for sensitive reporters of cellular activity. The list is subdivided into prokaryotic and eukaryotic applications. We have included the uses of luciferases in elucidating the control of gene expression or for monitoring cell viability. We have not included papers cited by Stanley and Stewart (J Biolumin Chemilumin 1990; 5:141-52) nor have we included papers on the structure and regulation of luciferases as this now substantial literature will be the subject of a future review. References cited in both this review and previous ones are referred to by the number assigned to them in the earlier review. PMID- 8237471 TI - Distribution of noncollagenous proteins in the matrix of adult human bone: evidence of anatomic and functional heterogeneity. AB - The microanatomic distribution of several noncollagenous proteins (NCPs) in bone matrix was examined by immunohistochemical analysis of glycol-methyl methacrylate embedded normal adult human bone biopsies. Osteopontin and bone sialoprotein stained throughout the lamellae of both trabecular and cortical bone. Cement lines (cortical and trabecular) and the mineralized matrix immediately adjacent to each Haversian canal were intensely stained. Osteocalcin was detected in cement lines; however, lamellar staining varied depending on the location within the individual unit of bone. In cortical bone, the inner concentric lamellae of osteons were often unstained but the outer lamellae were heavily stained for osteocalcin. Osteonectin was not detected in cement lines and in most specimens revealed a pattern similar to that of osteocalcin with respect to the absence of immunostaining within the inner concentric lamellae. Decorin was prominent in the perilacunar matrix, the canaliculi of osteocytes, and the matrix immediately adjacent to quiescent Haversian canals. Biglycan appeared evenly distributed throughout cortical and trabecular bone matrix. These results suggest that the incorporation of NCPs into matrix may vary depending on the stage of formation of individual bone units. The specific distribution and spatial relationship of these NCPs may be related to the function of each protein during bone resorption and formation. The distinct patterns of NCP localization in bone support the hypothesis that in addition to their structural and mineral-inducing properties, these proteins may influence the events associated with bone remodeling, such as recruitment, attachment, differentiation, and activity of bone cells. PMID- 8237472 TI - Post-translational control of collagen fibrillogenesis in mineralizing cultures of chick osteoblasts. AB - Cultured osteoblasts from chick embryo calvaria were used as a model system to investigate the post-translational extracellular mechanisms controlling the macroassembly of collagen fibrils. The results of these studies demonstrated that cultured osteoblasts secreted a collagenous extracellular matrix that assembled and mineralized in a defined temporal and spatial sequence. The assembly of collagen occurred in a polarized fashion, such that successive orthogonal arrays of fibrils formed between successive cell layers proceeding from the culture surface toward the media. Mineralization followed in the same manner, being observed first in the deepest and oldest fibril layers. Collagen fibrillogenesis, the kinetics of cross-link formation, and collagen stability in the extracellular matrix of the cultures were examined over a 30 day culture period. Between days 8 and 12 in culture, collagen fibril diameters increased from < 30 nm to an average of 30-45 nm. Thereafter, diameters ranged in size from 20 to 200 nm. Quantitation of the collagen cross-linking residues, hydroxylysyl pyridinoline (HP) and lysyl pyridinoline (LP), showed that these mature cross-links increased from undetectable levels to concentrations found in normal chick bone. Analysis of the kinetics of their formation by pulse-chase labeling the cultures with [3H]lysine showed a doubling time of approximately 5 days. The relationships between cross link formation, fibrillogenesis, and collagen stability were examined in cultures treated with beta-aminopropionitrile (beta-APN), a potent inhibitor of lysyl oxidase and cross-link formation. In beta-APN-treated cultures, total collagen synthesis was increased twofold, with no change in mRNA levels for type I collagen, whereas the amount of collagen accumulated in the cell layer was decreased by 50% and mineral deposition was reduced. The rate of collagen retention in the matrix was assessed by pulse-chase analysis of [3H]proline over a 16 day period in control and beta-APN-treated cultures. In control cultures, about 20% of the labeled collagen was lost from the cell layers over a 16 day period compared with > 80% in the presence of beta-APN. The beta-APN-treated cultures also showed a wider diversity of fibril diameters with a median in the > 45-60 nm range. In summary, these data suggest that cross-linking and assembly of collagen fibrils secreted by osteoblasts in vitro occur in a fashion similar to that found in vivo. The rate of cross-link formation is relatively constant and may be correlated with increasing collagen mass.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8237473 TI - Reduced creatinine clearance in primary osteoporosis in women. AB - We determined the relationship between bone mass and age, anthropometric variables, and serum and urine biochemical variables in 77 normal white women and 37 women with primary osteoporosis, 25 of whom had one or more vertebral compression fractures. Skeletal status was assessed by radiography of the hands with measurement of combined cortical thickness of the second metacarpal bones (CCT) or measurement of radial and lumbar bone density, or by both methods. Radial bone mineral content (RBMC) was measured by single-photon absorptiometry (SPA) and lumbar bone mineral density by dual-photon absorptiometry (DPA). Serum and urine biochemical variables were measured on days 6 and 7 of a controlled diet. In this mixed population of normal and osteoporotic women, we confirmed the strong positive correlation between creatinine clearance (Ccr) and bone mass that we previously reported in normal women. Multiple regression analysis showed that the relationship between Ccr and bone mass of the radius and lumbar spine was independent of age and body stature. Ccr was significantly lower in the 25 osteoporotic women with vertebral crush fractures than in age-matched normal women, and Ccr had predictive value for bone mass in individual subjects. The basis for the relationship between Ccr and bone mass has not been established. We excluded diminished production of 1,25-dihydroxyvitamin D as a result of declining renal function as a possible mechanism. PMID- 8237474 TI - Does weight-bearing exercise affect non-weight-bearing bone? AB - It was previously reported that weight-bearing exercise increases femur periosteal formation in 3-year-old, 230 kg cross-bred sows. To determine if this weight-bearing exercise also stimulated non-weight-bearing bone, bone formation in the seventh rib from these same sows was measured histomorphometrically on the periosteal, endocortical, and osteonal envelopes. The sows were randomly assigned by body weight to basal (B, n = 6), control (C, n = 7), or trained (T, n = 7) groups. After 3 weeks of exercise adaptation, T walked on a treadmill for 17 weeks at 5 km/h, 20 minutes/day, 5 days/week, at 5% grade. Groups were sacrificed initially (B) or after 20 weeks (C and T). Periosteal mineral apposition rate (MAR) was calculated over 136 days. Osteonal and endocortical MAR were calculated over the 14 days prior to sacrifice. There were no effects of exercise on the periosteal or endocortical percentage double-labeled surface (dLS/BS), osteonal remodeling frequency (N.dL.On/B.Ar), or MAR in any bone envelope of the rib. In conclusion, weight-bearing exercise sufficient to stimulate periosteal formation in the femur did not activate formation in the rib. Bone response to weight bearing exercise appears to be specific to the loaded bones. PMID- 8237475 TI - Oscillations in inositol 1,4,5-trisphosphate and diacyglycerol induced by vitamin D3 metabolites in confluent mouse osteoblasts. AB - For the last 5 years, attention has focused on the nongenomic effects of 1,25 (OH)2D3, but considerably less is known about the mechanisms of the nonnuclear actions of 24,25-(OH)2D3. The present study examines and compares the rapid (5-90 s) effects of 100 pM to 10 nM 24,25-(OH)2D3, 10 pM to 1 nM 1,25-(OH)2D3, and 1 100 nM 25-OHD3 on the formation of inositol phosphates and lipids in confluent mouse osteoblasts. 24,25-(OH)2D3 and 25-OHD3 effects were dose dependent; those of 1,25-(OH)2D3 were dose dependent in a bell-shaped manner. The two dihydroxylated metabolites induced a multiphasic response in inositol 1,4,5 trisphosphate (IP3) formation with three stimulation peaks; the IP3 response to 25-OHD3 was monophasic. The amplitude of the IP3 response to 24,25-(OH)2D3 was greater and its oscillation period was slower than that induced by 1,25-(OH)2D3. The diacylglycerol (DAG) responses to secosteroids showed two stimulation peaks that appeared at different times depending on the secosteroid used. Pretreatment with neomycin totally inhibited the first DAG response; neomycin had no effect on the second peak of DAG induced by 25-OHD3, whereas it partially blocked the second response of DAG to 24,25-(OH)2D3 and 1,25-(OH)2D3. These data show for the first time that 24,25-(OH)2D3 can modulate phospholipid metabolism in confluent mouse osteoblasts as early as 5-10 s. The first pathway used by all three secosteroids is that of the hydrolysis of phosphatidylinositol 4,5-bisphosphate via phospholipase C activation, leading to the formation of the two second messengers, IP3 and DAG, since neomycin totally blocked the response. Thus, the action of these secosteroids on the osteoblast membrane may also implicate several steps of the phosphatidylcholine cycle, according to the metabolite tested. Finally, these data point to a direct interaction of vitamin D metabolites with specific membrane recognition moieties. PMID- 8237476 TI - Bone resorption activity of particulate-stimulated macrophages. AB - Particulate wear debris from bone cement or prosthetic components can stimulate macrophages to cause bone resorption in a dose-dependent manner. This bone resorption activity of particulate-stimulated macrophages is associated with increased levels of both prostaglandin E2 (PGE2) and interleukin-1 (IL-1). In this study we compared the effect of particulate size, concentration, and composition on the secretion of IL-1 and PGE2 by peritoneal macrophages and on the bone-resorbing activity of conditioned medium (CM) harvested from particulate challenged macrophages. Particulates (titanium, Ti; polymethylmethacrylate, PMMA; and polystyrene, PS) only with phagocytosable size stimulated peritoneal macrophages to secrete IL-1 and PGE2 in a dose- and time-dependent manner. Ti particles (1-3 microns) exhibited significantly enhanced bone-resorbing activity measured as 45Ca release. The maximum bone-resorbing response was observed at a concentration of 0.1% Ti (approximately 10-15 Ti particulates per cell), which also corresponded with the highest IL-1 levels measured in particulate-challenged CM. This was measured using either conditioned media from Ti-stimulated macrophages or in cocultures of calvarial bone and macrophages in the presence of Ti. Exogenous PGE2 and recombinant human IL-1 could significantly increase the 45Ca release; indomethacin (IM) significantly reduced both the spontaneous calcium efflux and active 45Ca release from in vivo labeled calvarial bones. However, IM and/or anti-IL-1 antibodies could suppress only partly the macrophage mediated bone resorption, indicating that, in a macrophage-bone coculture system, factors other than PGE2 and IL-1 also may regulate particulate-induced bone resorption, probably involving multiple cell types. PMID- 8237477 TI - In vivo effect of 1,25-dihydroxycholecalciferol on the proliferation and differentiation of avian chondrocytes. AB - A combination of immunocytochemistry and in situ biochemistry has been used to determine the in vivo effects of 1,25-dihydroxycholecalciferol [1,25-(OH)2D3] on the proliferation and differentiation of chondrocytes. Chicks were fed a diet supplemented with 1,25-(OH)2D3 (2.5, 5, or 10 micrograms/kg diet) for 3 weeks, and measurements were made in sections of growth plate of chondrocyte proliferation and rate of maturation through the growth plate [using bromodeoxyuridine (BrdUrd) labeling] and also chondrocyte differentiation [assessed by alkaline phosphatase (ALP) activity]. The labeling indices of the control and supplemented chicks were similar (23.1 +/- 1.3 versus 23.2 +/- 1.6%); however, within a 21 h period the BrdUrd-positive cells of the supplemented chicks had moved down the growth plate significantly farther than in the control chicks (71.0 +/- 2.8 versus 52.6 +/- 1.8%). Greater ALP (mean integrated absorbance) activity higher up the growth plate of the supplemented chicks indicated a more differentiated phenotype in cells closer to the epiphyseal junction. Within individual transitional chondrocytes ALP activity in the 10 micrograms/kg supplemented chicks was 26.6 +/- 0.85, which was significantly higher (p < 0.01) than that of the control chicks (19.2 +/- 0.9). These results suggest that 1,25-(OH)2D3 in vivo does not increase the rate of chondrocyte proliferation but accelerates the onset of maturation. PMID- 8237478 TI - Training increases the in vivo strength of the lower leg: an experimental study in the rat. AB - The effect of training on bone strength has been investigated in rats. After 7 weeks of training, fracture strength of the tibia in vivo during muscle contraction and after resection was assessed. A group of 30 male rats 11 weeks old were randomized to exercise on a treadmill, sedentary (ordinary caging), and inactivity (right-sided patellar tendonectomy) groups. The training group ran on a treadmill with a 10% inclination for 1 h per day. After 4 weeks the animals in all groups were anesthetized and the right lower legs loaded in three-point ventral bending until fracture during electrically induced muscle contraction. The contralateral tibiae were tested correspondingly after resection. Ultimate bending moment, energy absorption to failure, bending stiffness, and deflection were assessed for the in vivo and the resected tibiae. The body weight gain was 37% higher in the sedentary and 57% higher in the inactive animals than in the training group (P < 0.05), indicating a physiologic effect of the training. In the dissected tibiae there were no significant group differences in any of the mechanical parameters, indicating that neither training nor inactivity changed the structural capacity of the tibiae per se. In contrast, there were significant differences between the in vivo tibiae. Ultimate bending moment was 12% higher in the training group than in the sedentary and inactive groups (P = 0.03). Energy absorption in the training group was 11 and 12% higher (not significant) than in the sedentary and inactivity groups, respectively. Bending stiffness was 7 and 17% higher in the training group compared to sedentary and inactivity groups (P = 0.018).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237479 TI - Human parathyroid hormone (1-34) and (1-84) increase the mechanical strength and thickness of cortical bone in rats. AB - An anabolic effect on bone of intermittent parathyroid hormone (PTH) treatment has been found in patients with osteoporosis and also in experimental animals. Controversies exist, however, about whether the positive effect on the trabecular bone balance occurs at the expense of the cortical bone. We examined the biomechanical quality of cortical bone after intermittent treatment with different doses of PTH and, furthermore, compared the effects of PTH-(1-34) and PTH-(1-84). Groups of rats were treated with biosynthetic human PTH-(1-34) or PTH (1-84), 1.1, 3.3, 10, or 30 nmol/kg/day for 30 days. No changes in the body weights and no changes in the lengths of the femora were observed after the PTH treatments. The biomechanical properties were analyzed by means of a materials testing machine. A dose-related increase in the bending strength and stiffness of the femora was found, and this increase in mechanical strength corresponds with a 9-12% increase in the cross-sectional area of the femoral diaphyses. The deflection capability and energy absorption were not influenced by any of the PTH treatments. No differences were found between the effects of PTH-(1-34) or PTH-(1 84) on the biomechanical properties of the femora. Consequently, intermittent treatment with biosynthetic PTH-(1-34) or PTH-(1-84) increased the formation of cortical bone, and the biomechanical competence of the femora was found to be preserved. PMID- 8237480 TI - Immunohistochemical detection and northern blot analysis of estrogen receptor in osteoblastic cells. AB - Expression of estrogen receptor (ER) was studied in MC3T3-E1 cells (mouse osteoblastic cell line), HOS TE85 cells (human osteosarcoma cell line), and primary osteoblastic cells derived from mouse calvaria with immunohistochemical techniques. The staining of ER was readily detectable in MC3T3-E1 cells, HOS TE85 cells, and primary osteoblastic cells by using a monoclonal anti-ER antibody that recognizes the DNA binding domain of ER. The immunoreactivity was distributed in the cytoplasm as well as in the nuclei. 17 beta-Estradiol (10(-8) M) did not alter this staining pattern. The expression of ER was confirmed by Northern blot analysis using rat ER cDNA probe, which revealed a 6.5 kb band in MC3T3-E1 cells and a 6.2 kb band in HOS TE85 cells. The mRNA level of ER was not altered by 17 beta-estradiol (10(-8) M). The immunohistochemical studies showed that ER was not detectable in all cells but in a small population of each cell type. This study is the first report to demonstrate the presence of ER immunohistochemically, and our results suggest the heterogeneity of ER expression among osteoblastic cells. PMID- 8237481 TI - Demonstration of a difference in urinary calcium, not calcium absorption, in black and white adolescents. AB - Bone mineral density (BMD) of the forearm, lumbar spine, and femoral neck is greater in black than in white children. Studies were performed to determine whether differences in intestinal absorption of calcium or urinary calcium or both account for an assumed more positive calcium balance and greater bone mass in black children. Normal black and white boys and girls were admitted to a metabolic ward and given a constant daily diet containing 1000 mg calcium, 60% as calcium carbonate, for 2 1/2 days (study I) or 3 1/2 days (study II). Fasting blood and 24 h urine collections were obtained, and in study II, unidirectional fractional absorption of calcium (alpha) was determined with stable isotopes of calcium. It was found that (1) serum 25-hydroxyvitamin D (25-OHD) and urinary calcium were lower and serum 1,25-dihydroxyvitamin D [1,25-(OH)2D] was higher in black than in white children, and (2) alpha was higher in boys than in girls with no racial difference, and (3) there were significant positive correlations between alpha and urinary calcium in the blacks and in the black and white children together. It is concluded that (1) alpha is higher in boys than in girls and (2) a lower urinary calcium, not increased intestinal absorption of calcium, is the means for a more positive calcium balance in blacks that accounts for the racial difference in BMD. PMID- 8237482 TI - Influence of exercise on cancellous bone of the aged female rat. AB - Dual-energy x-ray absorptiometry and dynamic histomorphometry were used to examine the effect of treadmill exercise on the bone density and cancellous bone formation and resorption in the proximal tibia and fifth lumbar vertebra (L5) of the aged female rat. Female rats aged 14 months were divided into four groups: 8 controls and 10 exercised for a 9 week study and 8 controls and 9 exercised for a 16 week study. Exercise consisted of running on a flat-bed treadmill, 17 m/minute, 1 h/day, 5 days/week. Tibial metaphysis and L5 vertebral density of each rat were measured in the 16 week study by DXA at weeks 0, 9, and 16. Compared to the control group, a significant increase in bone density in both metaphyseal tibia and L5 vertebra was apparent at 16 weeks after exercise training (P = 0.046 and 0.025, respectively, by two-way ANOVA). Histomorphometric analysis showed that the trabecular bone eroded surface and the ratio of eroded to mineralizing surface in tibial metaphysis were significantly lower in the exercised than in the respective control group in both the 9 and 16 week studies. In L5 vertebra, these decreases by exercise were apparent only in the 16 week study. A significant increase in the bone formation rate was apparent in the cancellous bone of the tibia but not of the vertebra after 16 weeks of exercise (P < 0.05). The trabecular architecture (bone number and separation) of the L5 vertebra in the exercised rats did not differ from that of the controls in either study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237483 TI - Osteoblast-specific expression of the alpha 2(I) collagen promoter in transgenic mice: correlation with the distribution of TGF-beta 1. AB - To begin to assess the transcriptional mechanisms that regulate type I collagen gene expression in differentiating osteoblasts, we have sought to determine the minimal promoter sequences that confer osteoblast-specific expression to the alpha 2(I) collagen gene during murine development. Transgenic mice were generated harboring DNA constructs in which the -2000, -500, and -350 to +54 regions located upstream of the start of transcription were linked to the Escherichia coli beta-galactosidase reporter gene (LacZ). Histochemical staining using X-gal indicated that the -2000 lacZ transgene was strongly expressed in newly differentiated and fully functional osteoblasts at intramembranous and endochondral sites of ossification. The promoter was also active in osteocytes in regions of bone remodeling within alveolar bone. The temporal and spatial activity of this region of the promoter closely resembled the developmental patterns of expression of the endogenous alpha 2(I) collagen gene as determined by in situ hybridization. The cis-acting elements within the 500 and 350 bp segments of the alpha 2(I) collagen promoter also drove reporter gene expression in forming osteoblasts, although levels of transgene expression were not as marked as that seen with the 2000 bp promoter. Furthermore, the synthesis and secretion of TGF-beta 1 in osteogenic zones coincided with areas where the alpha 2(I) collagen promoter constructs were transcriptionally active. Since a nuclear factor 1 binding site present at -300 has been shown to mediate the effects of TGF-beta 1 on the alpha 2(I) collagen promoter, these data support a role for TGF beta 1 in the control of this gene during development. PMID- 8237484 TI - Vertebral fracture assessment using a semiquantitative technique. AB - The assessment of vertebral fracture by conventional radiography has been refined and improved using either semiquantitative or quantitative criteria. The inter- and intraobserver variability was determined for a semiquantitative visual approach that we routinely use in clinical studies for assessing prevalent and incident vertebral fractures. In addition, the semiquantitative approach was compared with a quantitative morphometric approach. The incidence and prevalence of vertebral fractures were determined in 57 postmenopausal women (age 65-75 years) by three independent observers. The radiographic basis for fracture definitions and the source of interobserver agreement for the semiquantitative technique. We conclude that the semiquantitative approach can be applied reliably in vertebral fracture assessment when performed using well-defined criteria. PMID- 8237485 TI - Flanking markers define the X-linked hypophosphatemic rickets gene locus. AB - X-linked hypophosphatemic rickets (HYP) is an X-linked dominant disorder characterized by decreased renal tubular phosphate reabsorption and consequent hypophosphatemia. The defect in tubular phosphate reabsorption is probably secondary to an unidentified humoral factor. Identification of the humoral factor and a full understanding of the pathophysiology of the disease await the identification of the HYP gene. Previously we demonstrated that DXS257 and DXS41 are flanking markers for the HYP gene. Two markers, DXS365 and DXS274, are tightly linked to the HYP gene, but investigators have been unable to determine whether they are centromeric or telomeric to the disease gene. Since tightly linked flanking markers are necessary prerequisites to obtain the gene by positional cloning techniques, we sought to determine the relative positions of these markers to the HYP gene by expanding our data base for linkage studies. We also investigated a new polymorphic probe for linkage to HYP to construct a more detailed genetic map around the HYP locus. Our data indicate that the markers DXS365, DXS274, and DXS92 are tightly linked to the HYP locus and suggest a locus order of Xtel-(DXS444/DXS315)-DXS43-(DXS257/DXS3 65)-HYP-(DXS274/DXS41/DXS92)-DXS 451- DXS319-Xeen. These results will facilitate attempts further to localize and clone the HYP gene. PMID- 8237486 TI - The combined management of cerebral arteriovenous malformations. Experience with 100 cases and review of the literature. AB - A series of 100 patients treated for a cerebral arteriovenous malformation (AVM) is presented. Patients were admitted between 1985 and April 1992. Two groups are considered: the first group including 52 patients treated before the availability of radiosurgery (1985-1988), and the second group including 48 patients treated after the availability of radiosurgery (1989-1992). AVM's were classified in five grades according to the Spetzler's Grading System. Three techniques of treatment were used: surgical resection, intravascular embolization (with cyanoacrylate), and radiosurgery (linear accelerator). These three techniques were used either alone or in association, giving four types of management: surgical resection alone, embolization and resection, embolization alone, and radiosurgery (alone, or after embolization, or after surgical resection). From 1989 on, the availability of radiosurgery was responsible for the decrease of the "embolization and resection" group, which until then was predominantly used as well for low-grade (I, II, III) as for high-grade AVM's (IV, V). Overall, for the low-grade AVM's, the treatment of choice was surgical resection (79% of cases), with pre-operative embolization in one-half of these cases; the other low-grade AVM's were irradiated, with various combinations. For the high-grade AVM's, the treatment of choice was intravascular embolization (95% of cases), either alone, or followed by resection (45%) or radiosurgery (9%). Results were evaluated in terms of deterioration following treatment, in five groups: no deterioration (59%), minor deterioration (20%), long-lasting deficit (10%), major deterioration (5%), and death (6%). Overall, results improved after 1989: favourable outcome (no deterioration and minor deterioration) increased from 67% to 90%. Results were not related to the patients' age. More favourable results were obtained for low-grade AVM's (93%) than for high-grade AVM's (60%). For the low-grade AVM's the evolution from 1989 on (favourable outcomes increasing from 89% to 96%) occurred with the lowering of the mortality rate. For the high-grade AVM's, the evolution from 1989 onwards (favourable outcome increasing from 46% to 78%) occurred with the decrease of the cases with deficits. The angiographic results were strongly related to the management: 95% of complete eradication after surgical resection and 5% only after embolization alone. Concerning the results in irradiated cases, the follow-up is not long enough. The review of the neurosurgical literature since 1972 demonstrates progressive modifications in the therapeutic attitude as regards AVM's. The surgical management which was predominantly used at the beginning gave way progressively to a combined management, with a combination of embolization, surgery, and lately radiosurgery.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8237487 TI - The echo-guided removal of cerebral cavernous angiomas. AB - Twenty cavernous angiomas were surgically removed after being localised by intra operative echography. 10 patients were males and 10 females, with an average age of 34 years. Four patients had had intracerebral haemorrhage; all the remaining patients presented with headaches or seizures. The size of the lesion ranged from a minimum of 2.5 to a maximum of 4.0 cm. Its localisation was subcortical in 14 cases, paraventricular in 4 and in another 2 originated from the floor of the IV ventricle. Echography showed a blackberry-like cavernous angioma, hyperechogenous with respect to the surrounding parenchyma; the lesion was well-defined in all cases due to the absence of perilesional oedema with a clear demarcation from healthy tissue. After operation, pre-operative cranial nerve deficits (brought on by haemorrhage) only persisted in the two cases where the cavernoma involved the floor of the IV ventricle. In all cases post-operative radiological investigation recorded the complete removal of the cavernous angioma, the diagnosis of which was invariably confirmed by histological analysis. The authors draw the conclusion that intra-operative echography is capable of localising cavernous angiomas, providing the surgeon with real-time guidance during microsurgical removal. It is also useful for establishing as to whether complete removal has been accomplished. PMID- 8237488 TI - Shunt-dependent hydrocephalus after subarachnoid haemorrhage and aneurysm surgery: timing of surgery is not a risk factor. AB - Early hydrocephalus is a risk factor of shunt-dependent late hydrocephalus (SDHC). In the CT era 1980-1990 we had 835 consecutive patients operated on because of aneurysm and subarachnoid haemorrhage (SAH); 294 had an early hydrocephalus and 67 finally required a shunt. There were 14 patients with normal early CT and SDHC, in all 81 patients needed a shunt (10%). Patients with shunt did worse, they were older (53 vs 49) than the non-shunted group and there was a female preponderance. Pre-operative Grade correlated significantly with the need for a shunt operation; no one in Grade I developed SDHC, incidence in Grades III and IV was high (18% and 10%, respectively). Location was important; in vertebrobasilar area 28% and in anterior communicating area 14% but in middle cerebral area only 4% of the patients had SDHC. The amount of cisternal bleeding correlated significantly with SDHC; in 155 patients with non detectable or minimal cisternal blood only one developed SDHC, with severe cisternal bleeding the incidence was 16%. Ventricular bleeding increased the risk of SDHC, but intracerebral haematoma did not. Timing of surgery had no correlation with the risk of SDHC. Postoperative complications, haematomas and infections increased the risk of late SDHC. Delayed ischaemia correlated with the risk, but so did the treatment with nimodipine. Severe bleeding was the common predictor for the risk of SDHC. Location of the bleeding and postoperative problems are the other major causes. Outcome is, however, not so gloomy; 54% of patients with SDHC are independent one year later. PMID- 8237489 TI - Prediction of cerebral infarction due to vasospasm following aneurysmal subarachnoid haemorrhage using acetazolamide-activated 123I-IMP SPECT. AB - Prediction of cerebral infarction due to vasospasm (VS) following aneurysmal subarachnoid haemorrhage (SAH) was investigated using acetazolamide-activated (A A) N-isopropyl-p-[123I]iodoamphetamine (123I-IMP) single photon emission computed tomography (SPECT) in 79 SAH patients. A-A SPECT was undertaken twice or more for one each patient by Day 18. Fifty-six (71%) of the 79 patients presented with reduction of cerebral vasodilatory capacity (CVC) on SPECT due to VS by Day 18. Of the 56 patients, 29 showed CVC by Day 8 (Group A), while the other 27 first showed CVC reduction between Day 9 and 18 (Group B). Cerebral infarction on CT was revealed by Day 18 in 15 patients (52%) of Group A and 3 (11%) of Group B. Of the 56 patients, 20 showed reduced CVC in watershed[s] (Type 1), 12 in a sole territory of the intracranial major arterial trunk (Type 2), and 24 in several territories or in a sole territory with distant watershed[s] (Type 3). Cerebral infarction on CT by Day 18 was revealed in one patient (5%) in Type 1, 3 (25%) in Type 2, and 14 (58%) in Type 3. Twelve (71%) of 17 patients belonging to both Group A and Type 3 resulted in cerebral infarction. These results suggest that early and extensive CVC reduction are significant factors responsible for cerebral infarction due to VS following SAH. Cerebral infarction can be reasonably predicted using A-A SPECT in SAH patients. PMID- 8237490 TI - Failed back surgery syndrome. Part II: Surgical techniques, implant choice, and operative results in 171 patients with instability of the lumbar spine. AB - 171 patients with an instability-related failed back surgery syndrome underwent internal fixation. The operative indication was based on specific diagnostic procedures such as anaesthesia of articular nerves and the trial external immobilization in a plaster jacket. Taking account of the different types of hypermobility/instability of the lumbar spine, different spinal implants were used leading to excellent, good, satisfactory, moderate and poor results in 87 (53%), 42 (26%), 23 (14%), 9 (6%) and 2 (1%) patients, respectively. PMID- 8237491 TI - Metastatic spinal cord compression--options for surgical treatment. AB - Fourty-three cases with metastatic spinal cord compression were reviewed post operatively to clarify the usefulness of the procedures concerning restoration of neurological function, and pain relief. Only patients with pathological spinal instability and neurological sequelae were included. Posterior decompression and stabilization was performed in all but six patients. All but four patients (91%) reported decrease of pain symptoms. Amelioration of neurological function was achieved in 58%. Re-establishment of walking ability was obtained in 57%. Post surgery life expectancy averaged 11 months. In patients with widespread metastatic disease and/or multi-level instability of the spine restriction to palliative dorsal procedures is sensible. Post-operative ancillary treatment is necessary. PMID- 8237492 TI - Stereotactic management of congenital midline cysts. AB - The treatment of congenital midline cysts remains a controversial issue. The stereotactic management of 27 patients (6 symptomatic cavum septi pellucidi/cavum Vergae, 6 suprasellar cysts, 5 intraventricular cysts, 4 parasagittal cysts, and 6 supracollicular cysts) is reviewed. In 23 patients stereotactic ventriculo cystostomy by catheter implantation (internal drainage) led to clinical recovery or improvement accompanied by decreased cyst size. In four patients the internal drainage was not sufficient and was therefore completed as a ventriculo-atrial shunt system. Three minor complications (bleeding, aseptic meningitis, catheter infection) led to no sequelae. The results suggest that stereotactic internal drainage of these benign lesions is a safe, minimally invasive and efficient procedure. PMID- 8237493 TI - Air-ventriculography provokes an anterior displacement of the third ventricle during functional stereotactic procedures. AB - The width of the third ventricle, the length of the anterior commissure-posterior commissure line (AC-PC line), the spatial position of the midplane of the third ventricle, and the co-ordinates of the AC, the PC, and of 17 brain targets in the thalamus, hypothalamus and pallidum, were assessed on a pre-operative stereotactic computed-tomography (CT) study and compared to measurements on intra operative air-ventriculography, using a non-invasive relocatable stereotactic frame. There were no significant differences in the length of the AC-PC line, in the position of the midsagittal plane of the third ventricle, or in the vertical or lateral co-ordinates of the AC, the PC and the cerebral targets, between measurements on CT and on air-ventriculography. However, the width of the third ventricle was significantly larger, and the spatial positions of both AC and PC were significantly more anterior on air-ventriculography than on the CT study. This anterior dislocation of the commissures was presumably due to the insufflation of air into the ventricles of patients being in the supine position during surgery. PMID- 8237494 TI - Silicon prevents post laminectomy epidural root adhesions. An experimental study in rats. AB - An experimental study was designed to test the efficacy of a silicon barrier around a nerve root in order to prevent post operative epidural root scar adhesions. In 32 Sprague-Dawley rats a lumbar nerve root was microsurgically exposed bilaterally. In 16 animals a silicon tube, prepared with a longitudinal cut along half of its wall, was placed around one nerve root, while the contralateral side served as control. In another group of 12 animals, an autologous subcutaneous fat graft was placed on the exposed root. A group of 4 animals served as the control group which had not been operated upon. No post operative neurological deficit was observed in any of the animals. All animals were sacrificed 60 days after the operation and a block of tissue including the nerve roots were resected bilaterally without removal of the silicon or fat graft. Longitudinal and transverse cuts of the roots were stained with Haematoxilin and Eosin and with Masson's trichrome collagen stain. The roots in the "unoperated" control group were clean of any scar tissue. In 13 out of 16 animals, silicon prevented scar formation around the root as opposed to scar adhesions around control root on the contralateral side and as compared to unoperated roots. In the silicon group, adhesions penetrated only through the longitudinal narrow cut edge of the tube. Fat did not prevent adhesions in 11 out of 12 animals. We conclude that a silicon barrier is an effective method preventing post operative epidural root scarring in rats. PMID- 8237495 TI - 100 Years syrinx-surgery--a review. PMID- 8237496 TI - Pathogenesis of syringomyelia. PMID- 8237497 TI - Experimental models of syringomyelia--personal observations and a brief look at earlier reports. PMID- 8237498 TI - First experiences with syringomyelia imaging using a trans-oesophageal ultrasound technique. PMID- 8237499 TI - Technique and limitations of percutaneous sonography in the evaluation of the contents of the spinal canal. PMID- 8237500 TI - Value of intra-operative sonography in syringomyelia. PMID- 8237501 TI - Intra-operative sonography in syringomyelia--technique, results, limitations. PMID- 8237502 TI - On the evaluation of cranio-cervical decompression: normalisation of the cerebral spinal fluid circulation in MRI? AB - 1) By means of "cine-MRI" a semi-quantitative analysis of CSF pulsations in the aqueduct of Sylvius can be carried out. 2) This investigation enables a qualitative evaluation of the cranio-spinal CSF flow phenomena. On the one hand this information is a valuable aid for neurosurgeons for diagnosis and on the other, cine-MRI progress controls allow an evaluation of the success of an operation in the sense of a quality control. PMID- 8237503 TI - The value of somato-sensory evoked potentials in the diagnosis of syringomyelia. PMID- 8237504 TI - Intra-operative monitoring during syringo-endoscopy: results of median nerve stimulated somatosensory evoked potentials in nine patients. AB - Intra-operative SEP monitoring was performed in nine patients with cervical syringomyelia undergoing myelotomy and syringo-endoscopy. Several cortical and subcortical SEP components were evaluated. Pathological latency shifts of brain stem P14 were seen in two patients; other SEP components did not worsen nor recover intra-operatively. The clinical and pathophysiological significance of the data is briefly discussed. PMID- 8237505 TI - Syringobulbia: a surgical review. AB - Syringobulbia is a term which is applied to brain stem symptoms or signs in patients with syringomyelia or hindbrain herniation. Syringobulbia clefts due to a dissection of the CSF under pressure from the fourth ventricle should be differentiated from the ascending syringobulbia which may occur from upward impulsive fluid movements in a syringomyelia. Clinical analysis suggests that the commonest correlation of bulbar features is with neither of the above mechanisms but with pressure differences acting downward upon the hindbrain with distortion of the cerebellum and stem, traction on the cranial nerves or indentation of the brain stem by vascular loops. A review of patients with bulbar symptoms in a neurosurgical practice with an interest in hindbrain herniation in the Midland Centre for Neurosurgery and Neurology (M.C.N.N.) is presented. PMID- 8237506 TI - Syringomyelia: syringo-subarachnoid shunt versus posterior fossa decompression. PMID- 8237507 TI - Surgical strategies and results in syringomyelia. PMID- 8237508 TI - Neurosurgical management of the syringohydromyelia-complex. PMID- 8237509 TI - Syringo-peritoneal shunting: results and problems in a consecutive series. PMID- 8237510 TI - MRI follow-up in syringomyelia. Observations from twelve cases. AB - In 12 patients syringomyelia was diagnosed using MRI. The underlying cause was a hindbrain hernia in 8 patients, a posterior fossa cyst in one case, and in 3 patients the underlying cause remained unknown. Syringo-subarachnoid shunting in 8 patients failed to decrease pain and the size of the syrinx, motor lesion and gait disturbance in the majority of patients. A cysto-peritoneal shunt in the patient with a posterior fossa cyst and a syringo-peritoneal shunt resulted in a decreasing syrinx and clinical improvement. PMID- 8237511 TI - Disappearance of syringomyelia following resection of extramedullary lesion. A contribution to the aetiological enigma of syringomyelia. AB - A case of platybasia associated with syringomyelia is presented where the syrinx regressed nearly completely after transoral resection of the dens. This course seems to be of considerable interest because it is hardly explainable by the common theories which try to elucidate the pathogenesis of syringomyelic cysts. PMID- 8237512 TI - Two cases of thoracic intervertebral disk herniation combined with circumscribed syringomyelia--coincidental or causative? PMID- 8237513 TI - Introduction of a score system for the clinical evaluation of patients with spinal processes. AB - To facilitate statistical analysis of the clinical course of patients with spinal lesions such as tumours, dysraphic malformations, inflammation, or syringomyelia, we have developed a score system for each of the following symptoms and signs: sensory deficits, dysaesthesiae, pain, motor weakness, gait, bladder and bowel function. For each a score between 0 and 5 is given. The grading is designed in such a way that levels of functional significance receive separate scores: 0 = no function, 1 = severely disabled, 2 = disabled, 3 = severely compromised but function preserved, 4 = slightly compromised, and 5 = normal function. In general, scores between 0 and 2 indicate insufficient functional capacity and an unsatisfactory condition. Scores between 3 and 5 represent levels of function which should be the aim of treatment. We have used this scoring system for evaluation of almost 500 patients with spinal lesions both retrospectively and prospectively with good results in terms of reproducibility and validity. PMID- 8237514 TI - Mortality study on multiple sclerosis in the province of Ferrara, northern Italy, 1968 through 1989. AB - A mortality study on Multiple Sclerosis (MS) was carried out in the province of Ferrara, Northern Italy, over the years from 1968 to 1989 (mean population 382,379 inhabitants) to outline the temporal trend of the disease in the residing population that can be regarded as a representative sample of the caucasians of Northern Italy. Given the difficulties in performing retrospective incidence studies over long time periods, the mortality rate was used as an indirect indicator of MS occurrence. Through a review of mortality tabulations with 340 345 ICD code and an intensive survey of all the MS cases, with successive check of the deceased ones at the general register offices of the study area communes, 56 MS patients who had lived and died in the province of Ferrara in the period 1968-1989 were selected with an average crude death rate of 0.67 per 100,000 per year (95% confidence interval: 0.51-0.87), 0.55 per 100,000 if adjusted to the Italian population. The death rate was stable over the considered time period with only a slight but insignificant increase in the last years of the survey. No differences were found among the rates from the 5 Local Health Units (USLs) in which the study territory is subdivided. The highest age-specific death rates were in the fifth and sixth decade of life and the average duration was 21.17 +/- 11.05 years. The results are consistent with a relatively stable MS risk in the population of the province of Ferrara and a homogeneous occurrence of the disease in the study territory. PMID- 8237515 TI - Sporadic juvenile amyotrophic lateral sclerosis. AB - A case of sporadic amyotrophic lateral sclerosis with onset at 18 years of age and death one year after is reported. Only 8 other cases with such an early onset (and the typical clinical and pathological features of ALS) have been reported in the literature. Resemblances between these cases and adult ALS are pointed out, but since all knowledge about their etiology is lacking, the relationship between them remains unclear. PMID- 8237516 TI - Nociceptive threshold in hypothyroid patients. AB - The purpose of this study was to measure the nociceptive threshold in hypothyroid patients by determining when the nociceptive flexion reflex of the lower limb occurs under percutaneous electrical stimulation of the sural nerve, given that this threshold is well correlated with pain sensation. Twelve hypothyroid patients and twelve control subjects participated in the study. In the case of the hypothyroid patients, the nociceptive flexion reflex (or RIII reflex) was measured before and six weeks after the onset of substitution treatment. The results clearly indicate that the nociceptive threshold of the patients with hypothyroidism was significantly higher than that of the control subjects. After six weeks of substitution treatment, the RII threshold return to normal. The analgesia observed in the hypothyroid patients in this study do not appear to be correlated with the blood TSH level. The possible mechanisms of these analgesic effects are discussed. PMID- 8237517 TI - Long term prognosis of asymptomatic carotid artery occlusion. AB - Twenty six asymptomatic patients with internal carotid artery occlusion had ultrasound follow-up for a mean period of 34.3 months. During follow-up 3 patients (11.5%) reported transitory ischemic attacks (TIAs), one of these (3.8%) occurred in the vascular territory ipsilateral to the occlusion. There were not strokes. Annual TIA rate was 4% for all vascular territories, and 1.3% for the territory ipsilateral to the occlusion. Annual mortality was 9.4%. Our data on stroke risk in patients with asymptomatic carotid artery occlusion show a good prognosis in the long-term. PMID- 8237518 TI - Ischemic cardiopathy and other heart diseases: differences in carotid atheromas examined by duplex scanner. AB - Differences of frequency and morphology of carotid plaques are studied in 39 patients with ischemic cardiopathy matched for sex and age with 39 patients with various cardiopathies and 78 controls without heart diseases. Ischemic patients have more plaques, and these are more extended along the vessels. The plaques of the other cardiopathies reach greater thickness and degree of stenosis. PMID- 8237519 TI - Long-term survival of low-grade astrocytomas of the cerebral hemispheres. AB - The authors have reviewed 6 patients operated on, with histologically proven diagnosis of grade-II astrocytomas who have survived more than 10 years even with many reoperations. Young age, pre- and postoperative neurological status, aspect of the tumor mass on CT and MRI, site of the tumor in non-high functional areas and extent of surgical removal are the most important variables in predicting length of survival. In young patients without severe neurological deficits and with astrocytic tumors located in non high-functional areas, very long survivals may be obtained even with many operations, when the interval between the recurrences is at least three years. PMID- 8237520 TI - Urticarioid manifestation in course of cerebral imported malaria. AB - The authors report a case of urticarioid manifestation in a patient with imported pernicious cerebral malaria. They discuss the pathogenetic mechanisms of the cutaneous manifestation and point out the clinical importance of their observation. PMID- 8237521 TI - Hydatid cyst of the lumbosacral spine with large pelvic mass. AB - A rare case of hydatid cyst of the lumbosacral spine, causing extensive destruction of the sacrum and a large pelvis mass, is reported; a cutaneous fistula from the pelvic cavity to the posterior lumbar region was also present. The patient was studied by computerized tomography and operated on by combined anterior and posterior approach. The sacral localization of the spinal hydatidosis and its extension into the pelvic cavity are unusual. CT and MR allow a good definition of the bone destruction and the abdominal and pelvic extensions. Radical removal of spinal hydatid cysts may be rarely accomplished, because of the extensive bone invasion, and multiple recurrences, requiring repeating operations, occur in most cases. PMID- 8237522 TI - [Heparin and ischemic stroke]. AB - In absence of careful controlled trials, heparin therapy of ischemic stroke is based today on the clinical experience and personal belief of the physicians. Moreover, the incomplete knowledge of physiopathogenesis of ischemic stroke adds further confusion on those nosographic conditions treatable with heparin. This work aims to report the opinions prevalent in the literature, together with personal convictions, so to supply the reader with a complete view of the scientific discussion on this argument. PMID- 8237523 TI - [Bladder carcinoma: I. Epidemiologic considerations in the Basque country]. AB - Review of several facts of epidemiological nature, from a study on bladder urothelial carcinoma cases detected over the period 1983-1990 in our centre, a reference hospital serving the Bilbao area. Certain data recently published by the Euskadi Cancer Register allowed us to establish some considerations related to the presence of this entity in our setting. Its incidence nears the highest rates in industrial countries. Age distribution, but no the men/female ratio, is similar to that in other settings. Overall, no difference was detected in sex related survival. But, we corroborate that the youngest groups have the best prognosis. Also, an analysis is made of the association between age at diagnosis and several clinical and histopathological parameters. PMID- 8237524 TI - [Bladder carcinoma: II. Associations between various clinico-pathologic factors]. PMID- 8237525 TI - [Antimicrobial prophylaxis in urologic endoscopic surgery: pefloxacin versus ceftriaxone]. AB - Prospective, randomized study in 96 patients undergoing urologic endoscopic surgery to assess the efficacy of two antimicrobials, pefloxacin and ceftriaxone, in the prevention of urinary tract infections. The disease most frequently prompting surgery was vesical tumour and prostate adenoma. All patients had sterile pre-operative urine. Drugs were given endovenously two hours prior surgery in one single dose. Prophylactic efficacy was measured through urine culture before removal of the catheters. Incidence of bacteriuria greater than 100,000 colonies/ml was 20% for the pefloxacin group compared to 15.2% for ceftriaxone. These differences were not statistically significant (Squared-chi test). These results show that pefloxacin is as effective as ceftriaxone in the prophylaxis of endoscopic urological surgery. A single dose schedule was not entirely suitable to reduce post-operative bacteriuria. PMID- 8237526 TI - [Inhibition of tumor cell growth by the action of high-energy sources]. AB - In vitro comparison of the cytotoxic action of high-energy shockwave and megavoltage radiotherapy on tumoral cells. The inhibitory action these two therapeutic approaches have on cell viability is determined both singly and jointly, through life staining exclusion and nucleoside uptake tests. High energy shockwaves have a cytotoxic action significantly greater than low- and medium energy levels of megavoltage radiation. The cytotoxic effect from megavoltage radiation is significantly greater than that resulting from high levels of high energy shockwave as shown by the life staining exclusion tests. However, the nucleoside uptake test shows that cytotoxicity from either type of energy is no significantly different. When measuring the cumulative cytotoxic action of high energy shockwaves plus megavoltage radiation, the level of cytotoxicity is greater than the individual inhibitory actions on cell viability, but the degree of cytotoxicity induced cannot be comparable to that produced by higher energy levels from both sources separately. PMID- 8237527 TI - [Transplantectomy today: indications and complications]. AB - Analysis of the indications for transplantectomy and its complications over a 12 year period. Over a total of 159 failure grafts, we performed 53 transplantectomies (33.3%). The percentage of complications was 16.9%. Three patients died (5.6%) during the more or less immediate post-operative period. In our experience, consistent and prolonged maintenance of immunosuppression should avoid the need for transplantectomy in a high percentage of grafts. Only 15 transplantectomies were made on 104 failure grafts after 6 months (14.4%). Our short but successful experience with embolization of 2 rejected grafts confirms the validity of this conservative alternative versus conventional surgical transplantectomy in selected cases. PMID- 8237528 TI - [Transcervical resection (TCR) in uterine pathology]. AB - Review of our experience in transcervical resection. We contribute a total of 25 patients aged between 53 and 82 (mean age, 69 years), which presented with repeat metrorrhagia. The metrorrhagias were related to perimenopausal hormonal disorders (14 cases), submucosal uterine myoma (6 cases) and diffuse leiomyosarcoma (5 cases). PMID- 8237529 TI - [Acute scrotum as clinical presentation form of testicular tumor]. AB - Testis tumours usually emerge in young patients as a painless increase in the testis size accidentally noticed by the patient. Contribution of one case of non seminomatous testis tumour which presented clinically as an acute scrotum. An exhaustive appraisal of the clinical data allowed to choose the most appropriate approach for the patient in an emergency situation. PMID- 8237530 TI - [Leiomyosarcoma of the vena cava: presentation of 2 clinical cases]. AB - Presentation of two cases of leiomyosarcoma of a cava vein, an infrequent retroperitoneal tumour. Both cases were treated by means of anatomical reconstruction by a PTFE prosthesis. Brief review of diagnostic and therapeutical choices. PMID- 8237531 TI - [Bladder paraganglioma]. AB - By definition, the term paraganglioma includes all the tumours deriving from the body's paraganglion system, which are therefore ranked as intra-adrenal (pheochromocytoma) and extra-adrenal or paraganglion. Vesical paraganglion represents about 10% of all extra-adrenal paraganglioma and can be functional and non-functional. When functional, the symptoms are those originated from the products synthesised in excess, usually occurring in a paroxismical fashion, in relation with urination, ejaculation and defecation. Contribution of one vesical paraganglioma accidentally found in a 71 year-old female patients undergoing tests to study the extent of a right kidney carcinosarcoma. PMID- 8237532 TI - [Rupture of the cavernous body: report of 2 new cases]. AB - Review of two new cases of fracture of the cavernous body, one treated with early surgery and the other one conservatively. In both cases, the patients preserved good erections after one year. PMID- 8237533 TI - [High-flow priapism: early diagnosis and treatment]. AB - Contribution of a new case of high-flow priapism as clinical expression of an arterial-cavernous fistula caused by perineal injury. The diagnostic sequence was blood gasometry by means of puncture-aspiration of the cavernous body to establish the 'high-flow' taxonomical diagnose, with negative response to the intracavernous injection of vasoactive drugs; it was followed by arteriography, showing that the clinical picture was the result of an arterial-cavernous fistula, allowing at the same time a superselective therapeutic embolization. Vascular permeability and erection recovery were confirmed by ECO-doppler at the two- month assessment. PMID- 8237534 TI - [Intradiverticular bladder tumors]. AB - Retrospective study conducted on 58 patients with vesical diverticulum seen in our unit between 1975 and 1992, of which a total of 11 (18.9%) patients had vesical tumours. Of these, 6 (10.3%) were intradiverticular and 5 (8.6%) extradiverticular. Sixty-six percent of patients with intradiverticular tumours also had prior or concurrent history of extradiverticular vesical tumours. The most frequent clinical presentation was gross haematuria. The abdomino-pelvic CT is the most sensitive morphological examination although in 33% cases it resulted in overstaging. Curative treatment was only possible in the 4 patients with urothelial tumours, in whom 3 partial cystectomies with pelvian lymphadenectomy (2 pT1 G2 and 1 pT3 G3) and 1 TUR (T1 G2) were performed. The 2 (33%) remaining patients had advanced locoregional epidermoid carcinoma (T4 N+). All patients with urothelial carcinoma are alive with follow-up ranging between 6 months for the one infiltrant case and 136 months for a surface tumour undergoing partial cystectomy. Prognosis for both epidermoid carcinoma was ominous with mean survival time of 9 months. Review of the literature and discussion of epidemiological, clinical, diagnostic, therapeutic and prognostic issues. PMID- 8237535 TI - [Primary bladder carcinoma on diverticulum. Clinico-pathologic study of 5 cases]. AB - This paper reviews our experience with the vesical cancer which develops on the diverticulum, a situation occurring in 1.5% of all vesical tumours in our setting. The condition's clinical features (diagnostic and therapeutic), usually detected in advanced stages, are emphasized. From a morphological point of view, histology of over half these tumours differs from the pure transitional one. There were two cases of pure transitional carcinoma, one small cells cancer, one squamous cells cancer and one with adenocarcinoma differentiating foci. Several sets of data, which include histological variability as well as presence of congenital diverticulum and urethral tumours, have led us to suspect the existence of morpho-genetical factors in the development of these neoplasias. However, the widely accepted carcinogenetic theory continues to be the most likely. PMID- 8237536 TI - [Renal involvement in Burkitt's lymphoma]. AB - Review of one case of Burkitt's lymphoma in a 60 year-old male diagnosed with a renal tumour based on the symptomatology, intravenous urography (IVU), ultrasound and CT. In spite of the chemotherapy applied, the patient died 31 days after hospitalization. Burkitt's lymphoma is an infrequent tumour in our setting and renal invasion is rarely observed. The tumour's nature is neither revealed by IVU or ultrasound, CT being the most appropriate imaging technique showing a mass of decreased density that surrounds and infiltrates the tumour. The therapeutical approach for renal tumours is the same as for Burkitt's lymphoma. Besides the rarity of Burkitt's lymphoma, our patient was an atypical case for the age of presentation, early stage of renal invasion and fulminant evolution. PMID- 8237537 TI - [Bladder fibroma. Report of a case]. AB - Analysis of one case of benign vesical primitive connective tumour, with pathoanatomical description of vesical fibroma. The interest of this case rests on its rarity and uncertain origin, still subject to controversy. The paper presents the tumour's clinical history, pathological anatomy, therapeutical approach and prognosis. PMID- 8237538 TI - [Prostatic adenocarcinoma presenting as retrovesical cystic formation. Report of a case]. AB - Presentation of one case of prostate adenocarcinoma with a peculiar evolution towards a large cystic formation at the expense of the left prostate lobe, clinically revealed as a picture of poor performance status and impaired consciousness secondary to obstructive renal failure. The benign evolution of the case, even though the patient received no specific therapy, is emphasized. PMID- 8237539 TI - The value of the chest X-ray in making the diagnosis of bronchial asthma. AB - Although bronchial asthma is one of the most common chronic illnesses of children and young adults, it remains underdiagnosed. To assess the value of the chest X ray in helping to make the diagnosis of asthma at the primary care level, we studied the medical records of 58 patients aged 18-40 with the diagnosis of mild to moderate asthma and for whom both simple spirometry-forced one-second expiratory volume (FEV1) and/or peak expiratory flow rate (PEFR)--and a chest X ray had been performed. Only 21 of 58 (36%) had spirometry indicative of asthma (SPI+) while 34 of 58 (59%) had abnormal chest X-rays (CXR+)--"increased markings" and/or "low diaphragm." Although CXR+ discriminated between asthmatic patients and a normal control group, no difference was found between asthmatic patients and a group of patients with acute bronchitis. Nonetheless, the number of asthmatic patients with CXR+ and SPI - (n = 23) was significantly larger than the number with CXR- and SPI+ (n = 10), which indicates that for mild asthma the chest X-ray may be more sensitive than spirometry even though not as specific. These results were surprising at the time of the investigation. Subsequently, however, the importance of the inflammatory response in asthma came to light, which rendered the results more interesting than surprising. We conclude that the chest X-ray has value in making the diagnosis of mild bronchial asthma. PMID- 8237540 TI - An ecological approach to evaluating a special education program. AB - This study provides further evidence of the value of ecobehavioral measurements in program evaluations, especially since past evaluations have focused on individual-level factors rather than ecological factors. Static (e.g., classroom materials) and dynamic (e.g., group interactions) features of the classroom environment were used to describe the impact of a special education program on 84 behaviorally disturbed adolescent males. The program outcomes assessed included increases in appropriate and decreases in inappropriate behaviors. The findings reveal that the program apparently accomplished its objectives. However, the ecobehavioral data also suggest that these students were mostly involved in busywork; that is, they were working by themselves with instructional materials rather than participating in interactive activities with the teacher or peers. Ecological issues related to increasing student motivation are addressed. PMID- 8237541 TI - Attitudes of Mississippi college students toward David Duke before and after seeing the film Who is David Duke? AB - The attitudes of 211 students at a university in Mississippi were investigated both before and after seeing the Public Broadcasting Film Who Is David Duke? The film provided evidence of Duke's current racism, anti-Semitism, and pro-Nazi leanings. In a previous study with university students in Louisiana, the majority did not change their attitudes after watching the film (Eisenman, 1993). However, in the present study, students' attitudes showed change in an anti-Duke direction. The findings are discussed and reasons given for the differences between the two samples, and for the popularity of Duke, a former leader of the Ku Klux Klan. PMID- 8237542 TI - The ability of adolescents to predict future outcome. Part I: Assessing predictive abilities. AB - Because adolescents are generally viewed as "risk-takers," an assessment of planning and decision-making skills during the teenage years may reveal why this developmental group tends to engage in dangerous behaviors. One area in which these abilities have significant consequences for future outcome is pregnancy, a condition that requires a decision to either give birth or abort. Previewing, a process by which caregivers introduce infants to the physical sensations of imminent developmental change and the implications of such change on their relationship, may affect the adolescent's ability to predict upcoming change. This paper explores previewing skills during the adolescent years and how this capacity affects the teenager's orientation toward future outcomes. PMID- 8237543 TI - Cognitive, psychosocial, and reported sexual behavior differences between pregnant and nonpregnant adolescents. AB - A wide variety of variables have been related to the occurrence of adolescent pregnancy. However, many previous studies have produced conflicting results and are univariate in nature. The purpose of this study was to assess differences in pregnant and nonpregnant adolescents on variables from three domains: cognitive, psychosocial, and reported sexual behavior. Sixty-nine pregnant adolescents and 58 comparison adolescents filled out nine questionnaires presented on microcomputers. Significant differences were found on 10 of 24 univariate tests. The strongest differences concerned areas of scholastic functioning and reported sexual behavior; pregnant teenagers were more likely than nonpregnant peers to be doing poorly in school and less likely to use contraceptives. In addition, pregnant teenagers were more likely to have a relative or friend who was an adolescent mother and to expect child rearing to be easier than did the nonpregnant adolescents. A discriminant analysis was computed which correctly classified 83% of the sample, based on variables from each of the three domains. This study has served to replicate, refute, and expand on previous findings concerning the antecedents of teenage pregnancy. More importantly, this study has empirically demonstrated the multivariate and interrelated nature of variables associated with teenage pregnancy. PMID- 8237544 TI - Level of cultural mistrust as a function of educational and occupational expectations among black students. AB - The relationship between trust of whites and academic and occupational expectations of black students was explored. Black high school students were administered the Two-Factor Index of Social Position and the Cultural Mistrust Inventory. All students were also asked to indicate which occupation they expected to enter and the educational level they expected to attain. No differences were found between educational expectations and level of mistrust. However, students with lower occupational expectations had higher levels of mistrust. The implications of these findings are discussed. PMID- 8237545 TI - Transition-marking behaviors of adolescent males at first intercourse. AB - This study examined male transition-marking behaviors from adolescence into adulthood at the time of first intercourse. These include alcohol and drug use and sexual intercourse, and the companion behaviors of contraceptive use and planning of intercourse. The major findings were similar to those the authors described in a previous study as typifying female adolescents. Alcohol use at first intercourse was unrelated to use of contraceptives at that time, but was inversely related to whether the first intercourse was planned (i.e., those who did not drink were much more likely to plan). Planning was also positively related to contraceptive use, but the relationship was less strong than for female cohorts. PMID- 8237546 TI - Risk assessment for human immunodeficiency virus among pregnant Hispanic adolescents. AB - Inner-city adolescents are believed to be at increased risk for HIV infection because of their high-risk behaviors, and black and Hispanic teenagers may be at greatest risk as a consequence of these behaviors and inadequate AIDS education. In the present paper, the HIV risk status of pregnant Hispanic adolescents presenting for prenatal care to an inner-city municipal outpatient clinic located in New York City was assessed. The assessment consisted of a confidential structured interview. Based on known adult risk factors, adolescents were identified as being at increased risk or low risk. One-third of the 87 teenagers were identified as being at increased risk for HIV infection. Nobody reported symptoms suggestive of AIDS, had had a transfusion, or reported use of intravenous drugs or crack cocaine. Sexual risk-taking behavior was the most common factor that increased HIV risk. Sixteen adolescents were at increased risk solely because of a sexually transmitted disease and seven others reported an STD with at least one other risk factor. Although 86% of the pregnant teenagers reported the risk-taking behavior of substance use by self or partner, only four were at increased risk based on this factor alone. However, many adolescents were unaware of their partners' past substance use and sexual history and, therefore, may have underestimated their own risk. Birthplace (United States vs. foreign born) and nationality (Puerto Rican vs. non-Puerto Rican Hispanic) were significantly associated with the adolescents' HIV risk assessment via an interaction effect. None of the adolescents had previously considered themselves to be at increased risk for HIV infection. The overwhelming majority who were at increased risk declined referral for further counseling or testing. It is recommended that pregnant Hispanic adolescents be assessed for HIV risk and receive risk reduction counseling as part of their general health care. PMID- 8237547 TI - Adolescents and AIDS: knowledge and attitude. AB - Ninth- and tenth-grade adolescents' AIDS knowledge and attitude were assessed before and after a mandated AIDS education program. A control group of adolescents not exposed to the education program was included. As a result of the AIDS education, knowledge gains were indicated for the girls, but not for the boys. Both boys and girls gained in tolerance for AIDS victims. In turn, for the boys, but not for the girls, the knowledge gained had a direct effect on increasing tolerance toward AIDS victims. A plateau effect with regard to knowledge about AIDS is indicated. Gender differences and implications for future research on adolescents and AIDS education are discussed. PMID- 8237548 TI - Correlates of bulimia nervosa: early family mealtime experiences. AB - The relationship of early mealtime experiences to later bulimia was investigated in a sample of 128 female college students. Significant group differences were found between bulimics, nonbulimics, and repeat dieters on an early meal experience questionnaire, with the bulimic group reporting the most negative and unusual experiences. Significant differences were also found between groups on depression but not on family adaptability or cohesion. These findings suggest that bulimic, repeat dieter, and nonbulimic females have differentiating histories regarding eating-related issues in the family context. PMID- 8237549 TI - Lifestyle values of adolescents: results from Minnesota Heart Health Youth Program. AB - As a part of the Minnesota Heart Health Program (MHHP), a seven-year cohort study of adolescents in two different communities was conducted. A school-based intervention was implemented in one of the communities which addressed aspects of cardiovascular health promotion and risk-factor prevention. This paper focuses on changes in the adolescents' values and the importance of their behaviors and lifestyle patterns over the study period. Physical appearance was found to be the most valued characteristic of adolescents in both communities, the only value which grew in importance over time. The least valued behavior was the amount of TV they were allowed to watch. Students who participated in the intervention community tended to retain their positive values about physical exercise, whereas the reference community demonstrated gradual reductions. Girls in the intervention community tended to value the kinds of food they eat to a greater extent than did girls in the reference group. The study data might contribute to the search for more meaningful incentives in future preventive programs. PMID- 8237550 TI - Suicidal thoughts among adolescents: an intercultural approach. AB - Two epidemiological surveys of suicidal ideation were conducted among adolescents, aged 15-18, in France and the Canadian province of Quebec. The results suggest that, in both countries, suicidal ideation was linked to drug use (especially tobacco, illicit drugs, and psychotropic medicine), nonspecific somatic complaints (especially tiredness, sleep difficulties, depression, and feeling tense), a lack of self-esteem (pessimism, instability, boredom), and dissatisfaction with family relationships. There was no relationship between suicidal ideation and sports activities in either country. The implications for prevention are discussed. PMID- 8237551 TI - Cognitive correlates of depressive symptoms in hospitalized adolescents. AB - This study examined the relationship between self-reported depression and cognitive style in adolescent inpatients. Attributional style, perceived control over environmental events, and hopelessness were assessed in 80 consecutive admissions to an adolescent psychiatric unit. BDI scores were used to classify 36 patients as depressed and 44 as nondepressed. Adolescents who reported depression also reported significantly more internal attributions for negative events and less internal attributions for positive events, evidenced a more external locus of control, and described themselves as significantly more hopeless than did the nondepressed adolescents. Hopelessness and internal attributions for negative events were the strongest predictors of depressive symptoms. In contrast to the findings of most studies using clinical populations of depressed children and adolescents, a maladaptive cognitive style and self-reported depression were highly, positively correlated. PMID- 8237552 TI - The relationship of psychological adjustment to perceived family functioning among African-American adolescents. AB - The Family-of-Origin Scale for adolescents (FOS) is a 40-item ten subscale instrument designed to assess perceived psychological health in the respondent's family. To date, the FOS has been used exclusively with white adolescents. The FOS was administered concurrently with the Langner Symptom Survey (LSS), a measure of adjustment, to 135 African-American adolescents. The LSS exhibited moderate, yet significant, correlations with the FOS: A total score, two superordinate dimensions, and six subscales. The results lend modest support for the construct validity of the FOS. PMID- 8237553 TI - Correlates of loneliness among midwestern adolescents. AB - The extent of loneliness among midwestern adolescents was investigated in relation to depression, self-esteem, family strengths, parent-adolescent communication, age, and gender. The study sample consisted of 156 adolescents, ranging in age from 11 to 18 years, who were attending public schools in four midwestern communities. It was found that the subjects had high loneliness scores, and that older adolescents were lonelier than younger adolescents. There was a significant relationship between loneliness and depression. Loneliness was negatively related to self-esteem, family strengths, and mother-adolescent communication. The implications of these results for delineating the underlying dimensions of loneliness in adolescents are discussed. PMID- 8237554 TI - Characteristics of adolescent felons in a prison treatment program. AB - The entire population of inmates in a prison treatment program for youthful male offenders was studied (N = 43). Based on information from individual and group psychotherapy and from official files, and verified with another therapist, the following was found to be true for more than 50% of the offenders: they had anti social, anti-authority values; did not know how to be anything other than be a criminal; had been subjected to physical, psychological or sexual abuse, usually by a family member; were members of a minority group, i.e., black or Hispanic; and these were gang members. Whites were less likely to be gang members, but those who were tended to be Skinheads or Stoners. The findings can be used to help professionals understand and work with inmates to alter their criminal lifestyle. PMID- 8237555 TI - The role of activity in adolescent development: a study of employment. AB - Adolescent work values have historically been studied within the adult work value framework. This paper explores the notion that adolescence is a developmental stage, and therefore adolescent work values should be seen within this context. Such a developmental approach is found to be useful in reconciling some of the discrepancies in the literature on adolescent work values. In addition, two variables that may help explain the development of adolescent work values: activity and time perspective, are discussed. PMID- 8237556 TI - Metacognitive aspects of moral reasoning and behavior. AB - This study explored the notion that the development of moral reasoning and moral behavior may be linked to metacognition. The awareness of moral processes involved in moral reasoning and behavior was examined in 139 adolescents in three age groups. A number of significant moral metacognition-reasoning-behavior correlations were found. Intercorrelations for all dependent measures increased by age. A factor analysis revealed several components of moral metacognition (person, task, and strategy variables), and these were significantly related to increases in moral reasoning and behavior scores. More important, a contingency analysis supported the notion that specific types of metamoral knowledge (e.g., understanding the purpose, scope, and requirements of moral action) are related to high and low moral reasoning and behavior. PMID- 8237557 TI - Reflective monitoring in lonely adolescents. AB - Deficient social skills may be a primary contributor to chronic loneliness during adolescence. Specifically, adolescents at risk of becoming chronically lonely may have trouble reflecting on the appropriateness of their behavior, and learning from their actions. This inability to monitor and reflect on behavior may promote the development of the social, affective, and behavioral problems characteristic of chronic loneliness. Hence, some instances of chronic loneliness may be understood in terms of a developmental psychopathological model which delineates their cognitive, behavioral, and affective components. Dysfunctional familial or peer group interactions in childhood may contribute to the development of a reflective monitoring deficit. PMID- 8237558 TI - Perceived parental actions and evaluations of the family and its members. AB - In the present study, 64 college students described how their parents acted toward one another and also evaluated their families and various family members. Many significant correlations were found. Perceptions of parents' actions toward one another were associated with respondents' ratings of their families and individual family members. Interestingly, for females, but not males, there was a strong relationship between perceptions of how their fathers interacted with their mothers and how their mothers interacted with their fathers. Such findings lend support to the notion that ongoing parental interactions, and not just traumatic events such as parental death or divorce, are related to how youth and young adults subsequently view their families and the individuals within them. PMID- 8237559 TI - The cytokines: physiological and pathophysiological aspects. PMID- 8237560 TI - Estrogen and progesterone receptor proteins in patients with breast cancer. PMID- 8237561 TI - Advances in human leukocyte antigens and transplantation. PMID- 8237562 TI - Glutathione S-transferases: biomedical applications. PMID- 8237563 TI - Analytical applications of chemiluminescence. PMID- 8237564 TI - Contraception in Germany: a review. AB - In this paper studies on current contraceptive use and attitudes toward various contraceptive methods in Germany are reviewed. Both the former West and East Germany are considered. On the basis of the existing studies it is difficult to have a clear impression of which methods are currently used in both countries and how contraceptive practice developed over time, due to methodological differences between studies. Nevertheless, it can be concluded that use of reliable contraceptive methods is widespread in both East and West Germany. In this respect East Germany differs considerably from other previously communistic countries. Attitudes were mainly assessed in West Germany, and with respect to oral contraceptives and IUDs great concern appeared to exist regarding the safety of both methods. As far as oral contraceptives are concerned, selective mass media attention toward possible side-effects of oral contraceptives may have contributed to this situation. PMID- 8237565 TI - Quality of care in family planning in Latin America. AB - The majority of family planning programs in Latin America have been implemented in response to the region's population growth and aimed mainly at obtaining quantitative results, leaving quality, from the user's point of view, in second place. Recent decades have witnessed an increasing concern for quality of care in family planning because most researchers and service providers began to realize that when contraceptive methods are delivered without maintaining high quality levels, continuation rates are usually low and the image of methods and services is impaired. This paper reviews the level of quality of services in family planning in Latin America following Judith Bruce's framework and proposes, some activities that may improve it. The analysis found severe deficiencies in almost all the elements mentioned by Bruce, mainly in choice, technical competence, and mechanisms for improving continuity. The authors propose that to improve quality, services should (a) generate top management commitment to quality, (b) stay close to the client/user, (c) improve work conditions of providers/health workers, (d) measure quality, and (e) reward quality. The authors' conclusions are that (a) better quality services lead to better program results, (b) high quality is cost effective, (c) managerial commitment is critical to improvement of quality, and (d) quality of care is not a luxury; it is necessary and possible in all settings. PMID- 8237566 TI - Quality of care in family planning: clients' rights and providers' needs. AB - Quality of care means that the needs of the clients in the context of their personal life should be the major determinant of the behavior of the providers and the goal of the programs. Since family planning has been recognized as a right of individuals and couples, quality of care can be focused as a right of the client. Ten rights of family planning clients have been outlined by the International Planned Parenthood Federation (IPPF) as follows. Rights to: information, access, choice, safety, privacy, confidentiality, dignity, comfort, continuity, and opinion. The responsibilities for quality of care, and therefore fulfilling the rights of the clients, are distributed throughout the whole family planning program, but those who are actually seen as most responsible are the ones who are in direct contact with the clients--the service providers. A strategy for quality of care cannot be realistic without recognising that service providers have their own needs which can be outlined as: training, information infrastructure, supplies, guidance, back-up, respect, encouragement, feedback, and self-expression. When fulfilling the rights of the clients and needs of the service providers, both technical and human aspects should be taken into account. PMID- 8237567 TI - Oral contraception in the former Czech and Slovak Federal Republic: attitudes and use. AB - This paper presents and discusses data on oral contraception from a national representative survey (n = 1072) on knowledge, attitudes and practice with regard to contraceptive use in the Czech and Slovak Federal Republic (CSFR). Only 7% of the women at risk of unplanned pregnancy used the pill. The low rate of use seems to be due to the general negative image of oral contraception, reinforced by the experiences of past users and opinions expressed by relative, friends, doctors, and the media. In particular, perceived psychological disadvantages have an important bearing on the decision not to use the method, whereas perceived advantages have hardly any impact. It is likely that this negative image is closely linked with the types of oral contraceptives that have been available in the country and also with the limited choice. Second- and third-generation oral contraceptives have only very recently become available. It is recommended that balanced information on the advantages and disadvantages of oral contraception be made available to doctors and to the general public on a large scale and through a variety of channels. PMID- 8237568 TI - Family planning in Italy. AB - In the past 5 years, four knowledge, attitude and practice (KAP) surveys on family planning have been organized by the Unit of Population Survey Methodology of the Istituto Superiore di Sanita' (National Institute of Health). These surveys show that during the past 20 years, use of reliable contraceptives in Italy has increased, particularly in the North, encouraged by the more open attitude toward sexual behavior, following legalization of the provision of information on contraception and abortion upon request, and other social and cultural changes. In 1989 and 1991, 25% of women in Central and Southern Italy were using the pill or an IUD. However, the percentage of women not using contraceptives was still high (26% in 1989, and 19% in 1991) and withdrawal was the most widely used method (31% and 33%). The data show large regional differences. The main reasons for not using contraception, in the 1989 study, were fear of side-effects and ignorance, especially in the youngest and oldest women. Those least likely to use modern contraception were the young, the ill educated and the nulliparous women. Lack of information seems to be the main problem in Italy. In fact, the 5044 women interviewed showed a medium level of knowledge of contraception, with only 63% who could identify correctly the fecund period in the menstrual cycle. However, their attitude toward the use of modern contraception was positive; 65% of the women interviewed said they would recommend the Pill or an IUD to a friend, and 10% of them would recommend condoms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237569 TI - Factors associated with the choice of Norplant by women attending subsidized clinics in the US. AB - US women attending family planning clinics in Texas, were surveyed to determine which characteristics, experiences, behavior and specific needs were associated with the selection of the contraceptive Norplant. Race, ethnic origin, age and education were unrelated to choice of method. Increased numbers of previous pregnancies, familiarity with the method, dissatisfaction with previous methods, and sources of information about Norplant were associated with its selection. There was significant variation among the clinics in the proportion of patients to which they dispensed the method. Therefore, clinic selection may have influenced the outcomes of contraceptive decisions. PMID- 8237570 TI - The importance of training and supervision in quality of care. AB - Training of family planning service providers requires identification and selection of service providers, knowledge of their functions in the overall health team, provision of job descriptions and assessment of training needs. These can assist in the definition of training objectives and the establishment of program content. Varied teaching methodologies and aids can enrich the program and maximize learning and the venue and facilities provided for training should be accessible, appropriate in setting (using primary, secondary or tertiary facilities as necessary) and should provide an adequate standard of care. Evaluation of training performance should be regular and ongoing, with final assessment leading to recognition of training. The program should be evaluated by trainers and trainees to assist in improving the quality of the training and competence of the trainers. Finally, supervision and monitoring of performance in the workplace can be achieved by direct internal or external supervision, by indirect supervision using standing orders or guidelines, and by evaluation by managers, service providers and clients. PMID- 8237571 TI - Vasectomy and prostate cancer. AB - Numerous epidemiologic studies of vasectomized men have been conducted over the last decade. The majority of the reports have been reassuring with regard to possible long-term health consequences following male sterilization. In February 1993, two articles suggesting a relationship between vasectomy and prostate cancer appeared in the literature. Following publication of these studies, the United States National Institutes of Health (NIH) organized a meeting of experts to review the most recent literature on vasectomy and prostate cancer. After considering the evidence from the aforementioned studies and the preliminary findings of three unpublished studies, the NIH panel concluded that there was insufficient evidence of a real association between vasectomy and prostate cancer. This article reviews current information concerning the relationship, and recommends that providers continue to provide vasectomy as a contraceptive option. The research community should continue to investigate the possibility of a relationship between vasectomy and prostate cancer. PMID- 8237572 TI - Family planning in Russia: experience and attitudes of gynecologists. AB - A survey was made of 375 Russian gynecologists. The questionnaire on family planning and contraceptives was distributed at the beginning of three local educational symposia. Almost all participants filled in the questionnaire on attitudes, knowledge and experience with family planning. The sample consisted of departmental specialists and heads of departments/clinics, working mainly at in- or out-patient women's health clinics. The mean age was 37 years; 83% were women, living in medium-sized or large cities (80%). Half of them had been working as a gynecologist for more than 10 years. Only 55% had been trained in family planning. The main reasons mentioned for the high abortion rate in Russia were the lack of education, non-involvement of male partner, and lack of modern contraceptives. Most of the gynecologists were in favor of special family planning clinics with special attention to services for the users. About half of the physicians knew how the pill works and estimated that 41% of women know that the pill contains estrogens. Sixty-two percent found that patients are badly informed about available contraception. Main sources of information on contraception were journals/books, colleagues and mass media. The majority reported having a directive style of patient counseling, and stated that parents should be informed of their teenagers' sexual experiences. The more experienced physicians with a training in family planning were better informed on contraception and showed a more patient-concerned attitude. It is concluded that health care providers should be the main target group of training and education in family planning, and need the support of Western European family planning organizations. PMID- 8237573 TI - Inhibition of ovulation by a triphasic gestodene-containing oral contraceptive. AB - The minimal effective dose of gestodene for inhibition of ovulation was studied in 30 female volunteers. Daily doses of 10 micrograms to 50 micrograms gestodene were given orally for 21 days. A control cycle prior to treatment and a treatment cycle were monitored for LH, FSH, estradiol, progesterone and cervical score. At a daily dose of 40 micrograms of gestodene, 6/7 volunteers exhibited inhibition of ovulation and 1/7 had a cycle with luteal insufficiency. Ovulation was inhibited in all volunteers on 50 micrograms gestodene, suggesting that the minimum dose required to inhibit ovulation was 40 micrograms gestodene. Cervical score was blunted even at 10 micrograms gestodene. Similarly, 20 volunteers were treated with coated tablets containing ethinylestradiol/gestodene at 30/50 micrograms for 6 days, 40/70 micrograms for 5 days and 30/100 micrograms for 10 days. This triphasic gestodene-containing preparation inhibited ovulation in all 20 females. In one cycle in which follicle development was observed only 43 pg estradiol/ml was secreted. Data from this investigation suggest that this triphasic gestodene-containing OC has a high contraceptive efficacy. PMID- 8237574 TI - Evaluation of risk of thrombosis during use of low-dose ethinylestradiol desogestrel oral contraceptive. AB - Thirty healthy young women, non-smokers and of normal weight, used a combined oral contraceptive consisting of 20 micrograms ethinylestradiol and 150 micrograms desogestrel for 9 cycles. Before and during the 3rd, 6th and 9th cycles of contraceptive use, the following parameters were measured: triglycerides, total cholesterol, HDL-cholesterol, apolipoprotein A and B, prothrombin time, partial thromboplastin time, fibrinogen, antithrombin III, protein C, plasminogen, antiplasmin, tissue plasminogen activator, platelet count, platelet aggregation, beta-thromboglobulin and platelet factor 4. The ratios of total cholesterol/HDL-cholesterol and apolipoprotein A/B remained constant or showed only a slight increase. The clotting/fibrinolytic balance showed a similar trend. There was however, an inconstant but significant increase in antithrombin III and protein C. Platelet count and platelet function parameters were unmodified. Hence the contraceptive induced no substantial changes in lipid balance or blood clotting, at least during the study period. PMID- 8237575 TI - Some methodological considerations of a progestin-only oral contraceptive study from a programmatic perspective. AB - Among the limited number of reports on progestin-oral contraceptives (POCs), noncomparative studies conducted at family planning clinics have been the most often reported. This research trend will probably continue. Generally, noncomparative studies address a number of practical issues on POC use, and such studies have made important contributions to the understanding of this contraceptive modality. However, some studies have presented results that are either difficult to extrapolate for potential users or conflict with other findings. This article reviews several studies and suggests ways to improve the study methodology. Well-conceived, -executed, and -analyzed noncomparative studies can cost-effectively address a number of practical issues of POC use that have not been adequately addressed. Additional topics with programmatic utility, such as the effect of local background variables on POC acceptability, and the place of POCs in the ever-increasing contraceptive spectrum, are also discussed. PMID- 8237576 TI - Factors influencing attitudes about Norplant contraceptive subdermal implant. AB - Improving and widening Norplant use requires an understanding of users' complaints. A study was conducted of factors influencing user attitudes, with the goal of improving service delivery. METHODS: The study used a questionnaire consisting of 26 queries. An interview pertaining to the attitude of 610 users about Norplant implanted 5 years ago or more, was carried out in villages surrounding Semarang City, Central Java, Indonesia. RESULT: Of the 610 Norplant users, 84.59% expressed positive attitudes; 13.93% were neutral and 1.48% expressed negative attitudes. Factors significantly affecting user acceptance were: low socio-economic status; low education; activities in social groups; young age; more experience and knowledge about Norplant; few or no complaints. CONCLUSIONS: Acceptance of Norplant by villagers, depends on the socio demographic status and the complaints of users. This information can be used as the basis for improving the service delivery system. PMID- 8237577 TI - Planned condom use by women with Norplant implants. AB - This research examines the use of condoms to protect from disease women who selected hormonal implants for contraception. After receiving contraceptive implants, subjects were surveyed regarding their past use of condoms and their plans for using them to inhibit the transmission of HIV and other sexually transmitted diseases in the future. Twenty-one percent of women in the sample had used condoms in the past, but did not plan to use them once they have the implants. While decreasing their risk for unplanned pregnancy, they and/or their partners can be expected to experience an increased risk of exposure to sexually transmitted disease. Contraceptive providers must emphasize to their patients the need for protection from disease, in addition to pregnancy, when prescribing contraceptive methods. PMID- 8237578 TI - Evaluation of liver function and lipid metabolism following Norplant-2 rods removal. AB - The effect of Norplant-2 rods on liver function and lipid metabolism was evaluated in 100 Singaporean women. As with the Norplant six-capsule, the only change noted in liver function was that of a significantly elevated bilirubin, which was within the normal clinical range for the local population. As regards lipid metabolism, the mean values for total triglycerides, cholesterol, LDL cholesterol and HDL-cholesterol seen at the end of five years of Norplant-2 rod use and six months postremoval were similar to the preinsertion mean. As a result, the HDL-cholesterol/total-cholesterol-HDL-cholesterol and LDL cholesterol/HDL-cholesterol ratio remained fairly constant. This indicates that the use of Norplant-2 rods does not directly contribute to cardiovascular risk. PMID- 8237579 TI - Evaluation of hemostatic function following Norplant-2 rods removal. AB - Women using the Norplant-2 rod system in Singapore have been shown to have a change in their hemostatic function. There is a decrease in vitamin K dependent factors II, VII and a tendency toward lowered fibrinolytic activity after use of Norplant-2 rods. There was also an increase in platelet number and accelerated platelet aggregation. These changes are similar to those seen with the Norplant six-capsule system and it would appear that both systems do not activate the coagulation system or enhance a state of hypercoagulation. On removal of the Norplant-2 rods at the end of 5 years, the significant changes seen in hemoglobin function observed with Norplant-2 rod use remained. PMID- 8237580 TI - IUD (TCu380A) insertion following induced first trimester abortion. AB - In this study, 110 women (study group) who received an IUD (TCu380A) immediately after induced legal first trimester abortion were compared with 90 women (group 1) who started another type of contraception following legal abortion. Additionally, 100 women (group 2) who had an interval IUD insertion, were compared with both. In the study group, 3 (2.7%) patients were found to have an infection which did not fulfill the diagnostic criteria described by WHO and were treated easily by relevant antibiotics. One of 90 patients (1.1%) in group 1 developed acute PID after induced abortion within 7 days. None of group 2 patients had either a suspicion of pelvic infection or infection within the follow-up period (3 months). Among the women in the study group, bleeding and pain were not found to be more frequent, intense or of longer duration when compared with the other groups. Spontaneous expulsion of an IUD occurred in 2 patients (1.8%) of the study group and in 1 patient (1%) of group 2. At the end of the follow-up period, the continuation rates of the study group, group 1 and group 2 were 96.3%, 86.6%, 99%, respectively. PMID- 8237581 TI - Effects of the antiestrogen anordiol on ovulation and endometrial histology in the rat. AB - The rat was evaluated as a model for the study of the folliculostatic properties of anordiol (2 alpha,17 alpha-diethynyl-A-nor-5 alpha-androstane-2 beta,17 beta diol) which have been previously demonstrated in the primate. The effects in the rat were similar to those in the monkey in that anordiol: 1) extended the period of time for ovulation to occur, and 2) was effective only when administered during the early preantral stage of follicular development. The effect was different in that anordiol resulted in more complete luteolysis of newly formed corpora lutea. The presence of cornified cells in the vaginal smears 2 days after treatment with anordiol was a result of luteolysis and an intrinsic estrogenic activity of anordiol. Ovulation occurred on the first estrus after treatment whether the animals received 13 treatments every 5 days or only a single dose of anordiol. The uterine responses were characteristic of an antiestrogenic action of anordiol. It is concluded that the rat is a reasonable model for studies of the mechanism(s) of action of anordiol. PMID- 8237582 TI - Stress, the hypothalamic-pituitary-adrenal axis, and immune function. PMID- 8237583 TI - Marijuana and head and neck cancer. PMID- 8237584 TI - Evidence for a cannabinoid receptor in immunomodulation by cannabinoid compounds. PMID- 8237585 TI - Cocaine and immunocompetence: possible role of reactive metabolites. PMID- 8237586 TI - Molecular mechanisms associated with cocaine-induced modulation of human T lymphocytes proliferation. PMID- 8237587 TI - Opioids, receptors, and immunity. AB - The results achieved by those seeking to determine whether opioids and other drugs of abuse can affect immunity are quite astonishing given the short period of time that research has focused on this area. Despite the fact that there is no longer any question that opioids produce a variety of effects on the immune system, the extent and significance of these changes in the drug-abusing population remains to be determined. Whether or not the findings in mice and in in vitro preparations can be extrapolated to man is not yet known. Of major significance is the question of whether the endogenous opioid system is involved in immunoregulation. Given the multitude of drugs taken by drug abusers and the varying patterns of drug administration, the significance of the findings in the literature is still an open question. However, it is only by continuing studies such as those discussed at this meeting that we will find the answers. PMID- 8237588 TI - Effects of cocaine on the respiratory burst of murine macrophages. AB - Cocaine is a central nervous stimulant with a major potential for abuse. It is used clinically as a local anesthetic and vasoconstrictor. The effects of cocaine on the immune system have not been studied in depth. In this study, we have investigated the effects of cocaine on the respiratory burst (RB) of murine macrophages (M phi). The RB was measured by determining the increase in chemiluminescence. Both peritoneal and alveolar M phi were isolated from cocaine exposed mice and saline-exposed controls. Cocaine was administered by the intraperitoneal, intravenous, and intramuscular route. Both peritoneal and alveolar M phi from cocaine-exposed mice showed an increase in chemiluminescence when compared with M phi from matched controls. This effect was seen as early as one hour after cocaine exposure and lasted for up to 48 hours. Intraperitoneal injection of cocaine metabolites did not affect the RB. Macrophages exposed to cocaine in vitro failed to respond by an increase in RB. These findings indicate that cocaine induces the production of reactive oxygen intermediates (ROI) and suggests possible changes in M phi functions. PMID- 8237589 TI - Cocaine facilitation of cryptosporidiosis by murine AIDS in male and female C57/BL/6 mice. AB - As cocaine may affect progression of the Human Immunodeficiency Virus (HIV) infection to Acquired Immune Deficiency Syndrome (AIDS), we used a murine model of AIDS (MAIDS) induced by LP-BM5 murine leukemia virus to examine cocaine's possible role as a cofactor for secondary parasitic infections. Dissimilarities between the sexes were observed both in the absence and presence of the cocaine. The retrovirus-infected female mice had a much higher rate of Cryptosporidiosis than the retrovirus-infected male mice. Female, but not male, retrovirus-infected mice showed approximately 20-fold more Cryptosporidium per villus section than controls. Compared to respective gender controls, male and female animals infected with the retrovirus infection manifested a heightened Cryptosporidium oocysts count regardless of cocaine treatment. Overall, female groups incurred a higher incidence of infection compared to respective male groups. To determine the role of cocaine, groups of male and female C57BL-6 mice of similar age were treated with cocaine for 4 weeks followed by termination. Cocaine synergized with retrovirus infection in female mice to cause a 30-fold increase in the number of oocyst present. The spleen size and weight of female mice was significantly greater than uninfected controls or male mice. However, due to the very slow progression to murine AIDS in the males, parasite resistance was retained, including in cocaine treated C57BL-6 mice. Thymus cell number in the retrovirus infected female mice decreased significantly in comparison to uninfected female controls. Continued resistance to the parasite in male mice and its loss in female mice was due to the rate of immunosuppression and thus development of retrovirus-induced murine AIDS. PMID- 8237590 TI - Ethanol-induced suppression of in vivo host defense mechanisms to bacterial infection. PMID- 8237591 TI - Alcohol, cytokines and immunodeficiency. PMID- 8237592 TI - Ethanol affects macrophage production of IL-6 and susceptibility to infection by Legionella pneumophila. PMID- 8237593 TI - Suppression by dietary alcohol of resistance to Cryptosporidium during murine acquired immune deficiency syndrome. AB - Significant immunological changes occur following LP-BM5 murine leukemia retrovirus infection as well as chronic alcohol consumption. Retrovirus infection which has proceeded to murine AIDS permitted persistent Cryptosporidium infection, while non-retrovirus infected mice were resistant. Dietary alcohol provided until the day before parasite challenge did not affect resistance in controls, but increased the numbers of oocysts in the feces of retrovirus suppressed mice. Mortality was significant in retrovirus infected mice, and exacerbated slightly by dietary ethanol, while all controls survived parasite challenge. The retrovirus infected mice had greatly reduced numbers of intestinal CD4+ T helper cells and IgA+ B cells, which may explain their loss of intestinal resistance. Clearly, the severely immunosuppressed animals with murine AIDS were more sensitive to alcohol consumption than uninfected controls. This suggests that alcohol can synergize with murine retrovirus infection to exacerbate loss of resistance to an opportunistic pathogen common in human AIDS patients. PMID- 8237594 TI - Enhancement of HIV-1 replication by opiates and cocaine: the cytokine connection. PMID- 8237595 TI - Small animal model of AIDS and the feline immunodeficiency virus. PMID- 8237596 TI - Current status and future prospects in the immunotherapy of human immunodeficiency virus (HIV) infection. PMID- 8237597 TI - Consequences of opiate-dependency in a monkey model of AIDS. PMID- 8237598 TI - Perinatal AIDS: drugs of abuse and transplacental infection. AB - The number of children infected by the human immunodeficiency virus type-1 (HIV 1) who develop the acquired immunodeficiency syndrome (AIDS) continues to increase. While some children become infected after birth and others at the time of parturition, a significant percentage are infected during gestation and there is a positive correlation between maternal illicit intravenous drug use and fetal HIV-1 infection. Drugs can contribute in, at least, four ways to vertical transmission of HIV-1. These four ways are divisible into 2 main categories that are comprised of both direct and indirect mechanisms. For example, drugs of abuse can have a direct effect on the maternal-fetal interface. Cocaine is associated with vasculitis. If this occurs as a placentitis or chorioamnionitis, it can alter the permeability of these barriers to maternal blood and increase the number of potentially infected inflammatory cells in this tissue and as a result in the fetus. Another direct mechanism wherein drugs of abuse can increase the probability that a fetus will become infected is via an inflammatory reaction such as a vasculitis in the fetus rendering it more susceptible to viral infection. Drugs can also affect the course of HIV-1 infection via indirect mechanisms. An example of this may be by modulating the female immune system. This effect can exacerbate the woman's immunodeficiency and accelerate opportunistic infections. For example, cytomegalovirus infection resulting in placentitis might facilitate fetal HIV-1 infection. Lastly, a similar type of indirect mechanism can be postulated for the fetus wherein its developing immune system can be adversely effected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237599 TI - Solid tumors in HIV-infected patients other than AIDS-defining neoplasms. PMID- 8237600 TI - Stress, endocrine responses, immunity and HIV-1 spectrum disease. PMID- 8237601 TI - Immune function and drug treatment in anti-retrovirus negative intravenous drug users. PMID- 8237602 TI - Multiple pathogens may induce growth factor cascade resulting in KS. AB - While lipopolysaccharide endotoxin is the most prominent inducer of the kinecascade (TNF alpha, IL-1, 4, 6, 8) that leads to shock and multiple organ failure, bacterial exotoxins and products of certain gram positive bacteria can induce the same end results. We theorize that more than one pathogen can induce the sequence of protooncogene activation and growth factor release that results in the formation of KS. If KS has its own unique viral etiology, this virus has not as yet been isolated or identified but we continue to search for it. However, it is entirely possible that these lesions do not have a single well-defined etiologic agent but are the result of multiple agents cooperating in a set sequence. An endogenous, or apathogenic exogenous, retrovirus may replace HIV for initiator growth factor induction in CD4 cells in the classical (Mediterranean) or iatrogenic disease; and other pathogens co-exist or sequentially replace each other in the African endemic disease; whereas an array of viral pathogens (prominent among them CMV) take over growth factor induction in endothelial cells proliferating in response to the initiator growth factor (oncostatin M) released from HIV-infected CD4 lymphocytes in AIDS-KS. PMID- 8237603 TI - Exposure to the abused inhalant, isobutyl nitrite, compromises both antibody and cell-mediated immunity. PMID- 8237604 TI - Natural killer cells and Cryptococcus neoformans. PMID- 8237605 TI - Immune alterations in chronic morphine-treated rhesus monkeys. AB - Both immune and neuroendocrine abnormalities have been documented in heroin abusers. We investigated immunocompetence of peripheral blood mononuclear cells (PBMs) among separate groups of rhesus monkeys that were drug naive or received morphine either infrequently (twice/week) or daily (3.2 mg/kg). Both infrequent and daily morphine-exposed monkeys showed a decrease (10%) in the percentage of CD4+ circulating lymphocytes and an increase (19%) in the percentage of CD8+ cells. However, monkeys exposed daily to morphine showed a 30% increase in the helper-inducer CD4+ lymphocytes, CD4+CD29+, compared to untreated controls. PBMs taken from animals exposed daily to morphine responded poorly to forskolin in the production of cAMP compared with cells obtained from untreated animals. However, cells from monkeys that received morphine infrequently had elevated levels of cAMP in response to forskolin. These results suggest relatively low doses of morphine have a significant effect on immunocompetence in rhesus monkeys which parallels the effects observed in the opioid-dependent human population. Therefore, rhesus monkeys will provide a beneficial animal model to study the effects of opioid compounds on immunocompetence during exposure, withdrawal and substitution drug therapy in opioid-dependent animals. PMID- 8237606 TI - Immunosuppressive effects of morphine on immune responses in mice. AB - Implantation of a 75-mg morphine sulfate pellet subcutaneously into mice of different strains and sexes caused profound immunosuppression of their spleen cell primary in vitro antibody responses to sheep red blood cells. No sex differences were observed. In mice of the C3H lineage, naltrexone blocked the immunosuppression. In mice in the C57BL/6J lineage, naltrexone was ineffective in blocking the effects of morphine and was itself suppressive. In beige C57BL/6J bgJ/bgJ mice, placebo pellets were also suppressive. The mechanism of the morphine-induced immunosuppression was investigated in C3HeB/FeJ mice. Addition of normal splenic macrophages to in vitro cultures restored immune responses, as did IL-1, IL-6 and IFN-gamma, suggesting that morphine-induced immunosuppression is due to a deficit in macrophage function. Morphine pellet implantation induced splenic atrophy. Whether suppression is attributable to decreased macrophage numbers or to decreased functional capacity of individual macrophages is currently under investigation. PMID- 8237607 TI - Morphine-induced modulation of immune status: evidence for opioid receptor mediation and compartment specificity. PMID- 8237608 TI - Morphine binding sites on human T lymphocytes. PMID- 8237609 TI - Marijuana and bacterial infections. PMID- 8237610 TI - Psychological stress, immunity and immune depression. PMID- 8237611 TI - Effects of marijuana on spleen lymphocytes from mice of different age groups. PMID- 8237612 TI - Syphilis and drugs of abuse. AB - In summary, treponemal infections in hamsters treated with THC were slightly enhanced in comparison with vehicle controls. A greater degree of enhancement was exhibited in rabbits; treponemes proliferated more readily during treatment with THC than in control animals. Contrary to expectations, this occurred even in lesions which were not developed fully, i.e., were not ulcerated. Thus, treponemes were present in aspirates earlier during infection and from sites which had received smaller inocula of treponemes in these rabbits than in controls. Lesions in control groups developed ulcerations earlier than did the THC-groups, but treponemes were too scarce to be seen in pre-ulcerative lesions in these animals. It appears that THC-treatment enhanced infection with T. pallidum. It may be that in the local skin lesion, macrophages which are vitally important in early host responses to treponemal infection may not have been functioning optimally and mediators of tissue damage may not have been produced and, therefore, ulceration was delayed in spite of enhanced infectivity of the treponemes. PMID- 8237613 TI - Serum proteins affect the inhibition by delta-tetrahydrocannabinol of tumor necrosis factor alpha production by mouse macrophages. PMID- 8237614 TI - Blastocystis in humans and animals: morphology, biology, and epizootiology. PMID- 8237615 TI - Immunology of leishmaniasis. PMID- 8237616 TI - Transport of nutrients and ions across membranes of trypanosomatid parasites. PMID- 8237617 TI - The biology of fish coccidia. PMID- 8237618 TI - Giardia and giardiasis. PMID- 8237619 TI - Who should have a PEG? PMID- 8237620 TI - An evaluation of the efficacy and safety of tetrahydroaminoacridine (THA) without lecithin in the treatment of Alzheimer's disease. AB - Seventy-nine patients with probable Alzheimer's disease were enrolled into a double-blind, placebo-controlled cross-over study to assess the therapeutic effect and safety of THA (tetrahydroaminoacridine; tacrine) without concomitant lecithin administration. Forty-one patients completed the trial which consisted of two 12-week treatment phases separated by a 4-week wash-out period. Twenty-six subjects were withdrawn during the active treatment phase, mostly because of elevated transaminases or cholinergic side-effects, and ten during treatment with placebo. Statistical analyses were conducted on two groups of patients; those completing the cross-over and those with at least one evaluation in the first treatment period. This latter analysis, using the last observation carried forward was used to approximate an intention-to-treat analysis. THA was favoured over placebo in all three primary outcome measures (MMSE, ADAS Non-cognitive Scale, and the Functional Life Scale), but the results did not reach statistical significance. THA was favoured over placebo in five of the seven secondary outcome measures, but for only two of these was statistical significance attained. In terms of a three-point or greater increase in MMSE score, three to four times as many subjects improved on THA as on placebo. PMID- 8237621 TI - Post-operative electrolyte imbalance: its incidence and prognostic implications for elderly orthopaedic patients. AB - In order to assess incidence and prognostic implications of post-operative electrolyte disorders for elderly patients, 180 patients aged 79.5 +/- 6.8 years, range 70-96 years, admitted to a geriatric-orthopaedic liaison service were prospectively followed from admission to discharge or death. The association of age, basic medical conditions, type of anaesthesia and perioperative complications with post-operative sodium/potassium imbalance and fatality was assessed by logistic regression analysis. The incidence of post-operative electrolyte imbalance and fatality was 15% and 8.8%, respectively. Electrolyte imbalance was independently predicted by spinal anaesthesia [odds ratio (OR) = 2, confidence limits (CL) = 1.24-3.19], multiple pathology (OR = 2, CL = 1.1-3.58), use of cathartics (OR = 1.76, CL = 1.05-2.91) and intra-operative complications (OR = 1.7, CL = 1.03-2.88). Death was predicted by electrolyte imbalance (OR = 2.32, CL = 1.21-4.43), post-operative noninfective complications (OR = 2.3, CL = 1.09-4.84) and age greater than 79 years (OR = 1.17, CL = 1.06-1.3). Post operative electrolyte imbalance is a marker of very frail medical status and a risk factor for death among elderly orthopaedic patients. PMID- 8237622 TI - The association of age with plasma arginine vasopressin and plasma osmolality. AB - We investigated the association of age with supine and ambulant plasma osmolality (pOSM) and arginine vasopressin (AVP). Twenty-eight healthy-status-defined subjects were studied. Following an overnight fast, blood samples were withdrawn after one hour supine rest and two hours' ambulation. Neither supine nor ambulant pOSM nor AVP varied significantly with age. The results suggest that in healthy subjects age does not influence plasma osmolality or AVP. PMID- 8237623 TI - The scope of neurosurgery for elderly people. AB - Patients treated by a single neurosurgeon over a period of 17 years have been reviewed in order to assess the volume and nature of neurosurgical work performed on patients aged over 65 years. Based on this data we report: (1) The age distribution of the principal neurosurgical conditions, drawing attention to those conditions which are over- and under-represented in elderly patients. (2) A steady increase in the number of patients over 65 admitted over the last 15 years. (3) The numbers and types of neurosurgical operations carried out on elderly patients at the present time compared with 10 years ago. (4) The outcome of first-time surgery for intracranial meningiomas in 144 patients analysed by age. It is clear that a large and increasing proportion of neurosurgical work is carried out on persons over the age of 65. Age by itself appears to be no bar to a good outcome after a major neurosurgical operation provided that the surgery is elective and is not accompanied by a diffuse neurological disturbance. PMID- 8237624 TI - Hypertensive Old People in Edinburgh (HOPE) Study: electrocardiographic changes after captopril or bendrofluazide treatment. AB - Hypertension is a major risk factor for stroke and heart disease in elderly people. Eighty hypertensive subjects with mild cognitive impairment, aged over 70 years were drawn from a community screening programme and randomized to either captopril 12.5 mg twice daily or bendrofluazide 2.5 mg daily in a double-blind trial. Subjects were excluded if they had previously received antihypertensive treatment or had significant cardiac disease. Electrocardiograms (ECG) were recorded and scored at baseline and after 12 and 24 weeks of treatment. Mean blood pressure fell significantly and equally in both treatment groups. Most ECG variables remained unchanged throughout the trial but a significant reduction in QRS duration from 0.08 to 0.07 seconds was observed with captopril at 12 weeks (p = 0.004) and 24 weeks (p = 0.002). Subjects on captopril also exhibited a significant improvement in overall ECG classification at 12 weeks (p < 0.05), although this was not sustained at 24 weeks (p = 0.076). Captopril is a suitable alternative to bendrofluazide as an antihypertensive drug for elderly people with regard to ECG detected cardiac complications. PMID- 8237625 TI - Detecting depression in elderly medically ill patients: the use of the Geriatric Depression Scale compared with medical and nursing observations. AB - In a study of 59 elderly medically ill in-patients, 35% were found to have significant depressive symptomatology, as detected by the Geriatric Mental Status Schedule (GMSS). Of two screening methods, the Geriatric Depression Scale (GDS) performed satisfactorily but detection by nurses was poor. Acknowledgement of depression in medical casenotes was low. Training of nurses might improve detection; otherwise a mood-rating scale such as the GDS should be incorporated into routine practice. PMID- 8237626 TI - The association of age and frailty with the pharmacokinetics and pharmacodynamics of metoclopramide. AB - We have investigated the association of age and frailty with the pharmacokinetics and pharmacodynamics of the conjugated drug, metoclopramide. Six healthy young, six healthy elderly (> 65 years), and six frail elderly (> 65 years) subjects were studied on two occasions, receiving 10 mg metoclopramide by intravenous bolus and orally, in random order. Blood and urine were collected for measurement of pharmacokinetic parameters. Liver volume was measured by ultrasound. Sedation and contentment were self-recorded on visual analogue scales. Liver volume was not significantly different in the three groups, nor was bio-availability of metoclopramide. Clearance was similar in the young and fit elderly but reduced in the frail elderly subjects when compared with the young (p < 0.05), both when expressed in absolute terms and per unit liver volume. There were no differences in percentages cleared as the free drug or as the sulphate or glucuronide metabolite within or between groups, suggesting that frailty can produce a general impairment of conjugation pathways. The frail elderly subjects reported more sedation after intravenous dosage than the other subjects, whilst only young subjects reported akathisia. This did not relate to pharmacokinetic differences and seemed therefore to reflect associated pharmacodynamic changes in specific receptor or target sites. PMID- 8237627 TI - The social consequences of surgical complications for patients with proximal femoral fractures. AB - A case-matched study was performed to assess the social effects of surgical complications of operative fixation of fractures of the proximal femur. Fifty-two patients aged over sixty-five, half with deep wound infection, the remainder with serious early mechanical problems requiring revision surgery to correct these, had their hospital fatality, length of stay, mobility and social dependency at admission and on discharge compared with patients free from such complications. Deep wound infection was associated with a high fatality and in the survivors a prolonged period in hospital, a marked decline in mobility and without exception a great increase in social dependency; patients having revision surgery had a longer hospital admission, considerable alteration in social circumstances but a little decline in mobility and no significant increase in fatality when compared with matched cases. PMID- 8237628 TI - Prevalence and change of cardiovascular risk factors among men born 1900-19: the Finnish cohorts of the Seven Countries Study. AB - In a 30-year follow-up survey of the Finnish cohorts of the Seven Countries Study in 1989, 470 men aged 70-89 years were examined in two rural areas of Finland, in the East and in the West. Life-style-related coronary heart disease risk factors were at high levels in both groups, but the difference between areas found in the same cohort in middle age had mostly disappeared or partially reversed. Mean levels of systolic/diastolic blood pressure, body mass index, fasting blood glucose, blood glucose after 2-hour oral glucose tolerance test, and plasma fibrinogen were higher in the West than in the East (p < or = 0.05), but serum thiocyanate which reflects smoking habits was higher in the East (p < 0.001). No significant differences between areas were found in mean levels of serum total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides, apolipoprotein A-I, apolipoprotein B, lipoprotein (a), plasma coagulation factor VIIc, and prevalence of current smoking. PMID- 8237629 TI - Health care for elderly people in developing countries: a case study of Thailand. AB - The elderly populations of many developing countries are increasing rapidly. These demographic changes are a direct result of the success of socio-economic development that has led to declines in mortality rates at all ages and reductions in fertility. However, an aged population might be expected to suffer with the problems of dependency and disability leading to an increased burden of disease. Adequate disability and life expectancy data are essential for planning health and social policy but are currently insufficient. Making best use of the limited resources available for health care will require an emphasis on training of professional staff and carers, but with integration of health care for elderly people with established health services, particularly the existing primary health care systems. Strategies aimed at developing separate services for elderly people should be avoided because of the high likelihood of producing services of low quality with an inappropriate hospital emphasis. PMID- 8237630 TI - Review: trials of withdrawal of diuretics. PMID- 8237631 TI - Treatment of elderly people with hypertension. PMID- 8237632 TI - Investigation of 'syncope'. PMID- 8237633 TI - Aluminium and Alzheimer's disease. PMID- 8237634 TI - Aluminium and Alzheimer's disease. PMID- 8237635 TI - Anatomy of the human temporomandibular joint. An updated comprehensive review. AB - This article is an in-depth review of the current knowledge on human temporomandibular joints (TMJ's). All aspects of joint anatomy are described with emphasis on adaptability to biochemical stimuli throughout life. Each TMJ is a pressure-bearing compound double synovial joint. TMJ's are unique in having a movement not only controlled by the morphology of the joint per se but also by the dentition at the other end of the lever system. During life the temporal, condylar and discal articular surfaces undergo remodelling. The synovium is an important joint component which contributes to nourish and lubricate the avascular surfaces and has bactericidal properties. The joint capsule has privileged relationships anteriorly with the lateral pterygoid muscle. This muscle has two heads that show functionally reciprocal activation. The exact insertion and role of the superior head of the lateral pterygoid muscle remains controversial. The disc has a very low coefficient of friction and compensates for the lack of congruence between articular surfaces. Biomechanically it is stabilized between condyle and articular eminence by its thick rim which has special viscoelastic properties. Apart from the viscoelastic deformations the other important mechanism of disc stabilization seems to be related with the structure of some of the disc attachments. PMID- 8237636 TI - [Temporomandibular joint symptoms in orthognathic surgery: a retrospective study]. AB - The records of 152 consecutive patients who underwent orthognathic surgery in the Division of Maxillofacial Surgery A.Z. St.-Jan, Brugge, between 1/10/90 and 1/10/91 were evaluated for pre- and postoperative temporo-mandibular joint (TMJ) symptoms. Fewer TMJ symptoms were found postoperatively, than preoperatively (14.4% versus 19.7%). In the mandibular retrognathism group there were twice as much TMJ symptoms preoperatively in the low and normal mandibular angle patient group than in the high mandibular angle group (26% versus 13%). After surgery, there was a decrease of TMJ symptoms in the low and normal angle patient group (83% improvement). In the high angle absolute mandibular retrognathism group however, more new TMJ symptoms were seen postoperatively (21%). Bimaxillary surgical correction of a high angle absolute mandibular retrognathism case may provoke condylar resorption. Advice for follow-up and suggestions to reduce this annoying complication are given. PMID- 8237637 TI - The value of cryotherapy in the management of trigeminal neuralgia. AB - Cryotherapy to 32 individual branches of the trigeminal nerve in 27 patients, who were followed up for 3 years, resulted in pain relief out-lasting return of sensation. After one year 65% of patients were pain free. In the treatment of trigeminal neuralgia, cryotherapy is an easy method with similar results but lower complication rate in comparison with other peripheral methods. PMID- 8237638 TI - Neodymium-YAG and CO2 lasers in treatment of pre-cancerous lesions of the oral cavity. AB - The oral mucosa is an area of premalignant lesions such as leukoplakias and lichens. This work analyzes results obtained from patients who are carriers of these premalignant lesions treated with 1,060 and 1,320 nm Nd-YAG and CO2 lasers. Certain immediate and mediate post-op symptoms such as pain edema, hemorrhage, recidivism and scar fibrosis were studied and favorable results were obtained with this therapy. PMID- 8237639 TI - Feeding trials with dairy cows and heifers around parturition. PMID- 8237640 TI - Basic mechanisms for absorption of short-chain fatty acids. PMID- 8237641 TI - Sodium transport in cattle proximal and distal colon and interaction with short chain fatty acids. PMID- 8237642 TI - Interaction between absorption of sodium and acetate across the rumen epithelium of cattle. PMID- 8237643 TI - The pH fluctuation in the rumen of lactating cows. PMID- 8237644 TI - Acidosis in dairy cows. PMID- 8237645 TI - A pilot study on health and appetite in beef calves fattened on pelleted lucerne versus concentrate ad libitum. PMID- 8237646 TI - Some results of the etiology of alkalosis in high-yielding dairy cows. PMID- 8237647 TI - Different forms of rumen dystonia in dairy cows. AB - Four naturally occurring main forms of rumen dystonia of alimentary origin were observed in cattle: acidotic conditions; microflora inactivity; acidosis, and alkalosis. The following disturbances in the metabolism of carbohydrates, protein and energy were observed: 1. Acidotic conditions in rumen--ruminal pH, total and protein nitrogen decreased, whereas the total amount of VFA and the reduction activity of bacteria increased. 2. Inactivity of rumen microflora--significant changes of pH of rumen fluid were not observed, but the total number and activity of infusoria decreased. Fermentation of glucose, digestibility of cellulose, and reduction activity of bacteria decreased, whereas the amount of non-protein nitrogen increased. 3. Acidosis--ruminal pH, reduction activity of bacteria, and total number of VFA decreased. The percentage ratio between VFA changed--acetic acid concentration decreased, the concentration of valeric and caproic acids increased. The amount of total and non-protein nitrogen increased. 4. Alkalosis- ruminal pH increased, reduction activity of bacteria, fermentation of glucose, and concentration of VFA decreased. The amount of total and non-protein nitrogen increased. This investigation of different forms of rumen dystonia of alimentary origin is believed to be useful for the development of more effective treating methods and measures. PMID- 8237648 TI - Dutch experiments related to milk fever prevention. PMID- 8237649 TI - Nutritional risk factors for milk fever in dairy cattle: meta-analysis revisited. PMID- 8237650 TI - Propylene glycol in prevention of bovine ketosis. PMID- 8237651 TI - Is nine grams of calcium enough for treatment of parturient paresis? PMID- 8237652 TI - Incidence of milk fever and ketosis in Norway. PMID- 8237653 TI - Association between incidence risk of milk fever and lactation number, breed and season. PMID- 8237654 TI - Reports on disease in 14 organic farms in Denmark. PMID- 8237655 TI - Is rumen osmolality of grain-engorged cows influenced by pre-experimental diet? PMID- 8237656 TI - Diseases and feeding in Danish dairy herds. PMID- 8237657 TI - Disease and feeding in Norway. PMID- 8237658 TI - Periparturient diseases in dairy cows in Norway. PMID- 8237659 TI - Facts on diseases and feeding in the national Swedish dairy herd. PMID- 8237660 TI - National report from Iceland. PMID- 8237661 TI - Dairy production and diseases in Finland. PMID- 8237662 TI - Dairy cattle husbandry in Estonia. PMID- 8237663 TI - Dairy animal husbandry in Latvia. PMID- 8237664 TI - The problems of dairy farming development in Lithuania. PMID- 8237665 TI - Somatotropin, insulin-like growth factor-I and the mammary gland in regulation of nutrient and energy metabolism during early lactation. PMID- 8237666 TI - Hormonal regulation related to the development of fatty liver and ketosis. PMID- 8237667 TI - Epidemiological methods in integrated herd health programs. PMID- 8237668 TI - Epidemiological methods for identification and management of risk factors for production diseases in the dairy cow. PMID- 8237669 TI - Reproductive disturbances related to feeding. PMID- 8237670 TI - Appetite and gastroenteritis in ruminants. PMID- 8237671 TI - Total thyroid hormones and cortisol in plasma of Estonian black and white and Estonian red cows. PMID- 8237672 TI - Susceptibility to pregnancy disease in ewes and its relation to gestational diabetes. PMID- 8237673 TI - Dietary buffers and anion-cation balance in relation to acid-base status, feed intake and milk composition in the dairy cow. PMID- 8237674 TI - Parturition disease complex of the high-yielding dairy cow. PMID- 8237675 TI - Saliva secretion and acid-base status of ruminants. A review. PMID- 8237676 TI - Measurement of human brain lithium in vivo by MR spectroscopy. AB - PURPOSE: To quantify lithium in the human brain. METHODS: A 7Li MR spectroscopy method was developed with special features for high precision including: a) sampling a large cerebral volume to maximize the signal-to-noise ratio; b) adiabatic excitation pulses to ensure uniform spin mutation; c) morphometric analysis of the MR images of the sampled cerebrum; d) a mathematical model derived from empirical data to correct for receiver inhomogeneity effects; and e) a long interpulse delay, to eliminate errors arising from uncertain T1 values. RESULTS: A theoretical precision of 5.2% and an accuracy of better than 7.2% in someone with a brain lithium level of 1.0 mEq per liter of cerebral volume and precision and accuracy of 6.8 and 8.6%, respectively, in someone with 0.5 mEq/L brain lithium was calculated. This level of precision was surpassed in phantoms and patients. Brain lithium in 10 patients treated with lithium carbonate varied from 0.52 to 0.87 mEq/L (mean = 0.58 mEq/L; SD = 0.17 mEq/L). Brain-to-serum lithium ratios varied from 0.50 to 0.97 mEq/L (mean = 0.77 mEq/L; SD = 0.14 mEq/L). Substantial variation in brain lithium was observed in patients with similar serum lithium. CONCLUSIONS: A highly reliable method to quantify human brain lithium by 7Li MR spectroscopy has been implemented. Unexpected variability in brain versus serum levels of lithium was detected in patients with bipolar disease. PMID- 8237677 TI - Measurement of psychoactive drugs in the human brain in vivo by MR spectroscopy. PMID- 8237678 TI - MR detection of brain iron. AB - PURPOSE: To provide further quantitative studies concerning the relationship with age between regional brain iron and T2 shortening. METHODS: a) Quantitative T2 calculations of eight anatomic regions (red nucleus, substantia nigra, dentate nucleus, corpus callosum, caudate, putamen, temporal lobe white matter, and frontal lobe white matter) from T2-weighted spin-echo images were performed in 60 patients aged newborn to 35 years. b) Quantitative brain iron concentrations were obtained in six of the eight anatomic regions (red nucleus, substantia nigra, dentate nucleus, corpus callosum, cauda, and putamen) using 13 autopsied brains (newborn to 78 years). Brain tissue from these six regions was digested with 0.6 N HCl-2.5% wt/vol KMnO4 for 2 hours at 60 degrees C. After centrifugation, 0.1 mL of an iron-chelating reagent (2 mol/L ascorbic acid, 5 mol/L ammonium acetate, 6.5 nmol/L ferrozine, 13.1 mmol/L neocuprine) was added and the absorbance was measured at 562 nm/L and compared with a standard curve with ferric chloride. c) The in vivo iron concentrations in tissue that were obtained were reproduced in four test tube phantom studies with ferric ammonium sulfate or ferrous ammonium sulfate dissolved in either deionized water or 5% agarose. T2 calculations of the phantoms were made with a single-section multiple repetition time, multiple echo time acquisition. RESULTS: a) Clinical T2 calculations--all eight anatomic regions showed a decrease with age in T2 value, beginning shortly after birth. During the first three decades, the T2 shortening was most significant in the region of substantia nigra. b) Quantitative brain iron--five anatomic regions but not the corpus callosum demonstrated an age-related increase in brain iron (1449.6 nmol/g for the red nucleus versus 261.8 nmol/g for the corpus callosum). c) T2 effect of iron in vitro--both the ferric and ferrous iron phantoms showed a decreased T2 value in the in vivo concentration range of iron obtained from the postmortem studies. The T2 shortening was most marked for the ferric phantoms. CONCLUSION: There is an age-related accumulation of iron in five regions of the brain, correlating with an associated decrease in T2 value that can be demonstrated in iron phantoms. Brain iron appears to contribute to the progressive decrease of T2 signal that occurs with aging. PMID- 8237679 TI - The relation between regional brain iron and T2 shortening. PMID- 8237680 TI - Detection of recurrent thyroid cancer: MR versus thallium-201 scintigraphy. AB - PURPOSE: To compare MR with thallium-201 scintigraphy in the follow-up of patients who have undergone thyroidectomy and modified radical neck dissection for differentiated thyroid cancer. METHODS: Both MR imaging and 201Tl scintigraphy were performed in 39 patients after surgery for differentiated thyroid cancer. Ten patients did not have recurrence; 29 patients had 51 recurrent tumors in the neck and mediastinum. RESULTS: Among 51 tumor sites, 39 sites of recurrence were detected by MR and 24 were detected by 201Tl scintigraphy. The true-positive rate, false-negative rate, true-negative rate, and false-positive rate in detecting recurrent thyroid cancer were 76.5, 23.5, 100, and 0%, respectively, for MR and 47.1, 52.9, 100, and 0%, respectively, for 201Tl scintigraphy. MR was more sensitive than 201Tl scintigraphy in detecting recurrent tumors (especially small metastatic nodes). On T2-weighted MR images, recurrent tumor was characterized by high signal intensity. Scarring in the postoperative thyroid bed was characterized by low signal intensity on both T1- and T2-weighted images. Abnormal accumulation of 201Tl in the postoperative thyroid bed was not observed. CONCLUSIONS: Although both MR images and 201Tl scintigraphy were effective in distinguishing recurrent tumor from postoperative fibrous tissue, MR images were more sensitive than 201Tl scintigraphy in detecting recurrent tumors. These results suggest that MR imaging is more useful than 201Tl scintigraphy in the follow-up evaluation of patients after neck surgery for differentiated thyroid cancer. PMID- 8237681 TI - MR evaluation of laryngohypopharyngeal cancer: value of gadopentetate dimeglumine enhancement. AB - PURPOSE: To investigate the value of gadopentetate dimeglumine-enhanced MR imaging in determining the extent of laryngohypopharyngeal cancer. METHODS: Unenhanced and contrast-enhanced T1-weighted, proton-density-weighted, and T2 weighted images from 24 patients with laryngohypopharyngeal cancer were reviewed and compared with the pathologic findings of resected specimens. In 18 patients, ex vivo MR images of the resected specimens were also compared with pathologic findings. RESULTS: Laryngohypopharyngeal cancer showed intermediate intensity on T1-weighted and proton-density-weighted images, high intensity on T2-weighted images, and moderate enhancement on T1-weighted enhanced images. Cartilage invaded tumor enhanced moderately, whereas unossified cartilage in contact with tumors but without tumor invasion showed no enhancement. Laryngohypopharyngeal mucosa enhanced intensely; endolaryngeal muscles enhanced less. CONCLUSIONS: Enhanced MR images were more useful in assessing the extent of tumors and the presence of laryngeal cartilage invasion than were proton-density-weighted and T2 weighted images. PMID- 8237682 TI - Madelung disease: MR findings. AB - Two cases of Madelung disease (benign symmetrical lipomatosis) are presented. The MR findings in this striking condition are demonstrated. Short-repetition time/short-echo time sequences nicely show the relationship of the cervical lipomatous accumulations to the airway and major neurovascular structures in the carotid spaces. Fat-suppression techniques add no additional information in the radiologic evaluation of these patients. PMID- 8237683 TI - Anatomy of the junction of the inferior petrosal sinus and the internal jugular vein. AB - PURPOSE: To evaluate the anatomy of the junction of the inferior petrosal sinus and the internal jugular vein. METHODS: Using a previously described classification system, we prospectively classified venous anatomy bilaterally in 135 of 136 persons consecutively undergoing inferior petrosal sinus sampling. RESULTS: Type IV anatomy, with no anastomosis between the inferior petrosal sinus and the internal jugular vein, was significantly less frequent in our series than in a previous series (1 versus 7%; P < .001). Venous anatomy did not differ significantly between the left and the right junctions or between men and women. Venous anatomy was symmetric in only 65% of subjects (86 of 133). We describe an uncommon variant anatomy, incomplete type IV, found in 4.5% of our subjects (six of 133), that may cause incorrect results of petrosal sinus sampling. CONCLUSION: Bilateral sampling of pituitary venous effluent can be accomplished by the methods described, despite the presence of either incomplete or true type IV venous anatomy. Bilateral petrosal sinus sampling is anatomically possible in 99% of persons. PMID- 8237684 TI - Risk factors leading to cerebral arterial rupture by intravascular balloon. AB - PURPOSE: To clarify what is safe use of balloons in interventional neuroradiologic procedures. METHODS: Critical parameter values of balloon inflation and cerebral artery dilatation and rupture were determined. Dimensions and internal pressure were measured for a variety of latex and silicone balloons during inflation in both unconstrained and constrained environments including glass tubes, cadaveric human cerebral arteries, and canine basilar arteries. RESULTS: For unconstrained inflation, pressures within balloons inflated to the recommended maximum volume ranged from 200 to 650 mm Hg. When constrained, pressures became much higher for the same injected fluid volume. Balloon dilatation until artery rupture occurred only for balloons with diameters greater than 2.5 times the unstretched vessel diameter. Balloon pressures at vessel rupture ranged from 1000 to 2000 mm Hg. CONCLUSION: Pressures within inflated balloons vary with balloon type, material, degree of inflation, and constraint. Constrained balloons have markedly higher internal pressures, which may lead to vessel rupture if balloons are much larger than the vessel diameter. PMID- 8237685 TI - Failure-avoidance analysis: a brief introduction for neuroradiologists. PMID- 8237686 TI - Spontaneous carotid cavernous fistula presenting only with cranial nerve palsies. AB - PURPOSE: To discuss the differences in angiographic findings between cases of spontaneous carotid cavernous fistula with and without the classical triad of symptoms (pulsating exophthalmos, bruit, and conjunctival chemosis). METHODS: With CT, MR, and angiography, we examined 12 cases of spontaneous carotid cavernous fistula, five of whom presented only with cranial nerve palsies. RESULTS: In the seven cases with the triad, the main venous drainage from the cavernous sinus was the superior ophthalmic vein. Only one or two veins drained the cavernous sinus, and cortical venous drainage was not present in any case. In contrast, all but one case with only cranial nerve palsies had at least three venous drainage routes from the cavernous sinus, including cortical venous drainage. CONCLUSION: For the diagnoses of spontaneous carotid cavernous fistula, it is important to know that some patients do not show the classical triad of symptoms. In such patients, early diagnosis and treatment are particularly important because cortical venous drainage and a consequent risk of hemorrhage are frequently present. PMID- 8237687 TI - Percutaneous transvenous embolization through the thrombosed sinus in transverse sinus dural fistula. AB - Each of two patients presented with a dural arteriovenous fistula involving the transverse sinus. The sinus was thrombosed proximal and distal to the dural arteriovenous fistula with the venous drainage being retrograde through cortical veins. An ipsilateral percutaneous transjugular approach was used in both cases and allowed recanalization of the thrombosed sinus and embolization of the dural arteriovenous fistula with coils. Complete cure was achieved in one patient and 95% reduction in arteriovenous shunting including elimination of the cortical venous reflux in the other. This technique avoided surgical exposure of the sinus. PMID- 8237688 TI - Development of sigmoid dural arteriovenous fistulas after transvenous embolization of cavernous dural arteriovenous fistulas. PMID- 8237689 TI - Double spinal dural arteriovenous fistulas: report of two cases. AB - We report two unusual cases of double spinal dural arteriovenous fistulas. Both patients had signs and symptoms commonly described in this condition, but selective angiography of the spinal cord demonstrated two separate dural arteriovenous fistulas with separate venous drainage for each fistula. PMID- 8237690 TI - Percutaneous injection of an alcoholic embolizing emulsion as an alternative preoperative embolization for spine tumor. AB - A hypervascular pheochromocytoma metastasis in the sixth cervical vertebra was embolized preoperatively by an intravertebral injection of an alcoholic embolizing emulsion, when transarterial embolization was considered too dangerous because of the possibility of inadvertent embolization of the intracerebral vertebrobasilar territory. PMID- 8237691 TI - Mitochondrial disorders: analysis of their clinical and imaging characteristics. AB - PURPOSE: Investigation of the clinical, imaging, and in vivo MR spectroscopy (MRS) characteristics of disorders of mitochondrial function. METHODS: Clinical, imaging (five CT and 20 MR examinations), and MRS (six studies in five patients) findings in 19 patients with mitochondrial disorders were retrospectively reviewed. Results were critically analyzed and, when applicable, compared with results in the literature. RESULTS: Patients included four with mitochondrial encephalopathy with lactic acidosis and stroke-like episodes (MELAS), two with myoclonus, epilepsy, and ragged red fibers (MERRF), two with Kearns-Sayre syndrome, seven with Leigh syndrome, one with progressive cerebral poliodystrophy (Alpers syndrome), and three with trichopoliodystrophy (Menkes disease). MELAS, MERRF, and Kearns-Sayre tended to occur in older children and adults, whereas Leigh syndrome, Alpers syndrome, and Menkes disease occurred in infants and young children. All diseases involved gray matter early in their course, manifest primarily as T2 prolongation, with the deep cerebral nuclei being involved more often than the cerebral cortex. When T2 prolongation was seen in the white matter (MELAS, MERRF, Kearns-Sayre, Leigh), the peripheral and retrotrigonal white matter showed early involvement. Patients with Menkes disease showed rapidly progressive atrophy accompanied by large subdural hematomas. Proton MRS showed an elevated lactate level in involved regions of the brain; the lactate peak disappeared in old areas of T2 prolongation. CONCLUSIONS: Mitochondrial disorders have a wide range of both clinical and imaging findings. Although no one set of findings is diagnostic of these disorders, the combination of deep gray matter involvement and peripheral white matter involvement in young adults or children should suggest the diagnosis, especially when associated with an elevated lactate level on proton MRS. PMID- 8237692 TI - Moyamoya syndrome in young children: MR comparison with adult onset. AB - PURPOSE: To clarify whether there were any differences in MR appearance between the childhood and the adult moyamoya syndromes. METHOD: We compared the cranial MR findings in four children under the age of 6 who had moyamoya syndrome with previously documented adult cases. RESULTS: Moyamoya syndrome in younger children exhibited a significant increase in cortical and subcortical infarction, and a decreased incidence of deep white matter infarction in the centrum semiovale and basal ganglia, in contrast to adult cases. There were no remarkable differences between these two groups of moyamoya cases with regard to the occlusive changes of the internal carotid and middle cerebral arteries, or to the flow void sign on MR. CONCLUSION: These differences in the sites and frequencies of infarctions between the childhood and the adult moyamoya syndromes observed on MR might reflect differences in the cerebral circulation. PMID- 8237693 TI - Spiral CT of the temporal bone in unsedated pediatric patients. AB - PURPOSE: To determine the feasibility of performing rapid, high-resolution, low radiation-dose spiral CT scans of the temporal bone in unsedated pediatric patients. METHODS: Axial spiral CT examinations with an effective section thickness of 1.25 mm were performed in 41 infants and children. Multiplanar reconstructions were generated using 360 degrees and 180 degrees linear interpolation algorithms. Three-dimensional reconstructions of the middle ear were also performed. RESULTS: Complete axial spiral scans were performed without sedation in all patients. Three hundred sixty-degree linear interpolation images had comparable resolution to conventional CT; 180 degrees linear interpolation images with an effective section thickness of 1.0 mm at 0.2-mm intervals showed good detail of middle and inner ear anatomy, and multiplanar reconstructions from this algorithm were comparable to direct scans. Three-dimensional reconstructions clearly displayed the oval window, malleus, and incus. CONCLUSION: Spiral CT is a clinically feasible method for rapidly evaluating the temporal bone in pediatric patients, eliminating the need for sedation. The images obtained from this technique are comparable to conventional CT. High-quality multiplanar reconstructions can be generated from the spiral data set, providing an alternative to direct scans in multiple planes. Additionally, three-dimensional images of internal temporal bone anatomy can be generated from the spiral data. PMID- 8237694 TI - Radiologic-pathologic correlation. Alobar holoprosencephaly. PMID- 8237695 TI - Leigh disease (subacute necrotizing encephalomyelopathy): MR documentation of the evolution of an acute attack. AB - The radiologic evolution of Leigh disease is documented with sequential brain MR in the acute phase of the illness, at 3 weeks, and at 3 months. High-signal intensity lesions seen on T2-weighted images in the first week resolved by 3 months, whereas new lesions appear during the chronic stage. Putamenal involvement is not a pathognomonic radiologic finding. Brain stem tegmentum, particularly the mesencephalon, is characteristically involved on MR in the early and late phases of the illness. PMID- 8237696 TI - Menkes kinky hair disease: characteristic MR angiographic findings. AB - We report two cases of Menkes kinky hair disease in which MR and MR angiography were performed. The clinical and imaging features are reviewed. MR demonstrated characteristic cerebrovascular tortuousity and thus may be a valuable aid in diagnosis and follow-up. PMID- 8237697 TI - Infantile Krabbe disease: complementary CT and MR findings. AB - This case report of the infantile form of Krabbe disease in a 2 1/2-month-old boy illustrates the complementary findings that may be seen on CT and MR scans. The key finding on the CT scan was increased density in a bilateral symmetrical distribution involving the thalami with extension into the centrum semiovale. The MR scan, on the other hand, more clearly showed demyelination in the brain stem and cerebellum. In cases in which this involvement is minimal or absent, the MR scan may fail to detect an abnormality, and a CT scan will, therefore, still be necessary to detect characteristic abnormalities. PMID- 8237698 TI - Olfactory neuroblastoma: MR evaluation. AB - PURPOSE: To evaluate MR in the diagnosis and staging of olfactory neuroblastoma. METHODS: Five patients with histologically proved olfactory neuroblastomas were studied by MR. Four also had CT scans. RESULTS: The extent of disease delineated by MR agreed with surgical findings and surgical staging of the tumor in all five patients. All cases of olfactory neuroblastomas originated in the nasal cavity and involved the ethmoid sinuses. In three of the five cases, the tumors extended to the anterior skull base. In one case MR correctly suggested skull base involvement missed on CT. Lesions were isointense to hyperintense with muscle on T1-weighted scans. Compared with fat on T2-weighted scans, olfactory neuroblastomas were variable in signal intensity. Enhancement was minimal to moderate in all cases. CONCLUSION: In the evaluation of olfactory neuroblastoma, MR is most useful in delineating the extent of the tumor and may be more accurate than CT. However, the signal intensity characteristics of olfactory neuroblastomas may overlap other tumors. PMID- 8237699 TI - MR of intraparotid masses. AB - PURPOSE: To determine which MR techniques are best for identifying intraparotid masses and to assess the utility of MR for predicting specific diagnosis. METHODS: A retrospective review of 31 intraparotid masses was performed. Lesion conspicuity and signal intensity characteristics were evaluated by objectively measured regions of interest and by subjective visual review. RESULTS: The T1 weighted images produced the highest lesion conspicuity in 25 of 31 masses. All the masses were easily detected as hypointense lesions on the T1-weighted scans with good or excellent conspicuity; on the T2-weighted images nine lesions appeared isointense to the normal parotid tissue, including six malignancies. Gadolinium-enhanced images were not helpful for identifying the intraparotid masses. Warthin tumors and malignancies tended to be hypointense or low in conspicuity on T2-weighted images. CONCLUSIONS: The T1-weighted images were best for detecting parotid masses. The T2-weighted images may help in limiting a differential diagnosis, but a specific diagnosis cannot be made without biopsy. PMID- 8237700 TI - Posttraumatic rhinolith. AB - Rhinolith, calcified concretion of a nidus within the nasal cavity, is a possible diagnosis for certain patients with densely mineralized lesions that appear benign and without bone destruction. PMID- 8237701 TI - The accuracy of CT and MR evaluation of the sella turcica for detection of adrenocorticotropic hormone-secreting adenomas in Cushing disease. AB - PURPOSE: To document the accuracy of CT and MR of the sella turcica for detecting adrenocorticotropic hormone-secreting adenomas in Cushing disease. METHODS: The radiologic findings of the sella turcica prior to transsphenoidal surgery are reviewed in 141 patients who had biochemical evidence of pituitary-dependent Cushing disease. Axial thin-collimation CT scans with sagittal and coronal reformations before and after contrast enhancement were obtained in 125 patients. Seventy-eight patients had MR examinations with a 1.5-T superconducting magnet. In 11 of the patients gadolinium-enhanced MR scans were also obtained. The preoperative interpretation of the imaging studies was correlated with the surgical findings and patients follow-up. RESULTS: The sella turcica was enlarged in 43 cases (30%). In 125 patients reformatted or direct coronal thin-collimation CT scans were available. Seventy-eight of the patients had MR. In the 12 patients with pituitary macroadenomas, the accuracy of CT (n = 10) and MR (n = 10) in respect to detection of the lesion was 100%. Of the 98 microadenomas assessed by CT, 47 (48%) were directly depicted as distinct hypodense lesions. In only 31 of 73 cases (42%), however, could CT predict the precise anatomic location and extent of the lesions. Only patients in whom the hypercortisolism was corrected by later surgery were considered for the correlation analysis. Of the 52 microadenomas assessed by MR, 28 (53%) were directly depicted as distinct lesions of reduced signal intensity on T1-weighted images, and in only 21 of 41 cases (52%) did MR show good correlation to the surgical findings. Some degree of partially empty sella was found in 22% of the patients. CONCLUSIONS: Although both the sensitivity and the diagnostic accuracy of imaging methods of the sella turcica have been considerably improved in comparison with previous reports, they still provide only a minor contribution to the diagnosis and differential diagnosis of Cushing syndrome. PMID- 8237702 TI - Intrasellar and suprasellar paraganglioma: CT and MR findings. AB - We present the CT and MR findings of an intrasellar and suprasellar paraganglioma in a 17-year-old girl. Paragangliomas in this location are rare. The lesion had several imaging features characteristic of paragangliomas including marked enhancement after contrast and multiple vascular flow voids. PMID- 8237703 TI - MR imaging of "spray heads": toluene abuse via aerosol paint inhalation. AB - PURPOSE: Three male patients with a history of spray-paint inhalation are presented. METHODS: Spin-echo MR was used to evaluate the central nervous system changes secondary to toluene inhalation delivered via spray-paint fumes. RESULTS: The remarkable findings included the loss of cerebral and cerebellar gray-white matter discrimination, scattered multifocal deep white matter lesions, and gross generalized atrophy of the cerebrum, the cerebellum, and the corpus callosum. CONCLUSION: Although the observed changes are nonspecific, combined with a positive history the diagnosis of inhalation toluene abuse can be made on the basis of consistent MR findings. PMID- 8237704 TI - Multiple sclerosis presenting as Parinaud syndrome. AB - The authors report a patient presenting with a 1-week history of paresis of upward gaze as his initial manifestation of demyelinating disease. They stress that: 1) multiple sclerosis can present as Parinaud syndrome (paralysis of upward gaze); and 2) it is important to rule out a non-contour-deforming intraaxial midbrain lesion if a posterior third ventricle or pineal region mass is not identified on screening studies in these patients. PMID- 8237705 TI - Fast spin-echo MR imaging of the cervical spine: influence of echo train length and echo spacing on image contrast and quality. AB - PURPOSE: To examine the interaction of echo train length and interecho spacing and their effects on image quality and contrast in fast spin-echo sequences of the cervical spine. METHODS: Forty-three patients with suspected cervical disk disease were prospectively evaluated with fast spin-echo with varying echo train lengths and interecho spacing. A flow phantom was used to confirm findings related to cerebrospinal fluid pulsation. Parameters were manipulated to adjust contrast, signal-to-noise ratio, the effects of artifacts, and the speed of acquisition. RESULTS: In general, increasing echo train length increased homogeneity and high intensity of cerebrospinal fluid signal and reduced acquisition time; however, it decreased the signal-to-noise ratio of cerebrospinal fluid and cord and increased blurring, and, to a lesser extent, edge enhancement, and "truncation-type" artifact. Increasing interecho space permitted the use of longer echo times but minimally decreased contrast and signal-to-noise ratio of cord and cerebrospinal fluid. In addition, increasing echo spacing increased blurring, edge enhancement, truncation-type, magnetic susceptibility, and motion artifacts. CONCLUSIONS: For cervical spine imaging, a long echo train length and short echo spacing partially compensate for cerebrospinal fluid flow and produce the best myelographic effect but must be modulated by other constraints, such as artifact production or technical capabilities. PMID- 8237706 TI - Lumbar degenerative disk disease: prospective comparison of conventional T2 weighted spin-echo imaging and T2-weighted rapid acquisition relaxation-enhanced imaging. AB - PURPOSE: To compare conventional T2-weighted spin-echo imaging with a rapid acquisition relaxation enhanced (RARE) technique in the routine evaluation of lumbar degenerative disk disease. METHODS: Thirty consecutive patients referred for evaluation of the lumbar spine for suspected degenerative disk disease were evaluated with sagittal and axial T1-weighted spin-echo, conventional T2-weighted spin-echo, and T2-weighted RARE "turbo spin-echo" sequences (4000/93/2 (repetition time/echo time/excitations), 192 x 256, echo train length of 8). Conventional T2-weighted and RARE images were evaluated independently by two neuroradiologists for image quality, presence of artifacts, cerebrospinal fluid signal intensity, extradural interface conspicuity, intradural nerve root conspicuity, soft-tissue detail, and signal intensity of normal and degenerated intervertebral disks. RESULTS: Both readers rated the cerebrospinal fluid signal higher, the extradural interface conspicuity higher, and the nerve root detail greater on the turbo spin-echo than on conventional spin-echo images. Neither reader had a significant difference in ranking "normal" or "degenerated" disk signal on the two sequences. Both readers rated soft-tissue detail higher for conventional than for turbo spin-echo. CONCLUSION: RARE sequences can replace conventional T2-weighted spin-echo sagittal studies for degenerative lumbar disk disease. PMID- 8237707 TI - Subacute combined degeneration of the spinal cord: MR findings. AB - The authors present the initial and follow-up MR findings in a patient with subacute combined degeneration of the spinal cord, a complication of vitamin B12 deficiency, and a rare cause of demyelination of the dorsal and lateral columns of the spinal cord. Initial study showed high intensity in the dorsal columns of the cervical and thoracic spinal cord on T2-weighted image. Five months after treatment, the abnormal signal intensity was noted to have decreased. PMID- 8237708 TI - Cerebellar arteries originating from the internal carotid artery. AB - PURPOSE: To study the incidence of cerebellar arteries originating directly from the internal carotid artery, and to discuss the probable embryological origin and clinical implications of this anatomical anomaly. METHODS: We reviewed 5500 angiographic studies done at our institution in the last 4 years, searching for these anomalous vessels as well as for other anatomical variants. RESULTS: Eleven cerebellar arteries originating directly from the internal carotid artery and nine embryonic cerebral arteries were found. CONCLUSION: These arteries are considered persistent trigeminal artery variants and seem to occur as the result of the persistence of a primitive trigeminal artery associated with an incomplete fusion of the longitudinal neural arteries. These anomalous vessels are clinically significant particularly because of their role in endovascular therapeutic and surgical complications and the paradoxical lesions in the cerebellum that occur as a result of carotid disease. PMID- 8237709 TI - Hippocampal anatomy and pathologic alterations on conventional MR images. PMID- 8237710 TI - Cystic acoustic schwannomas: MR characteristics. AB - PURPOSE: To evaluate the spectrum of MR characteristics of cystic acoustic schwannoma and to investigate its incidence. METHODS: We retrospectively reviewed the MR findings and clinical records of 16 patients with cystic acoustic schwannomas. In addition, the MR examinations of 411 consecutive patients referred for clinical suspicion of acoustic schwannomas were reviewed retrospectively to assess the incidence of acoustic schwannomas with cystic lesions arising from the internal auditory canal. RESULTS: Of the 16 acoustic schwannomas with MR evidence of intramural cysts, 11 tumors had single small cysts, and five had multiple intramural cysts of variable size. Intramural cysts in 11 of the 16 tumors exhibited higher signal intensity than that of cerebrospinal fluid; the remainder were isointense to cerebrospinal fluid on both T1- and T2-weighted images. All intramural cysts showed circumferential enhancement after contrast administration. Nine of the 16 cystic acoustic schwannomas also had MR evidence of extramural/arachnoid cysts. Six of the extramural/arachnoid cysts had epicenters away from the dural interface, and the other three cysts were broadly based against the dura. The incidence of cystic acoustic schwannomas was 11.3% and association with extramural/arachnoid cysts 7.5%. CONCLUSION: Our series suggests that cystic changes in acoustic schwannomas and the association with extramural/arachnoid cysts are not as rare as previously reported by other diagnostic methods. The high signal intensity of intramural cysts is probably related to necrotic material, blood, or colloid-rich fluid. The difference in the MR characteristics of extramural/arachnoid cysts associated with acoustic schwannomas and those of typical arachnoid cysts not associated with neoplasia may be related to higher protein and/or colloid contents secreted by the tumor. Most extramural/arachnoid cysts had epicenters between the tumor and brain, suggesting that the most likely mechanism of formation is peritumoral adhesions. It creates a pseudo-duplication caused by the trapping of fluid between the leptomeninges and the mass, resulting in an acquired type of arachnoid cyst. PMID- 8237711 TI - Wegener granulomatosis with meningeal involvement. AB - The clinical, radiographic, and neuropathologic features of the case of a 41-year old man with Wegener granulomatosis presenting with neurologic symptoms are correlated. CT and MR scans of the head demonstrated extensive meningeal thickening and enhancement. The importance of considering this diagnosis, confirmed by antineutrophil cytoplasmic autoantibodies, is emphasized. PMID- 8237712 TI - Neurilemmoma of the oculomotor nerve presenting as an orbital mass: MR findings. AB - Neurilemmoma of the oculomotor nerve is rare and infrequently presents as an orbital mass. We report a neurilemmoma arising from the cavernous sinus with its major part extending into the orbit. The Antoni A part of the neurilemmoma showed intermediate signal intensity in both T1- and T2-weighted MR images and exhibited postcontrast enhancement. The Antoni B part revealed hypointensity on T1-weighted images, hyperintensity in T2-weighted images, and showed no contrast enhancement. PMID- 8237713 TI - Epidermal nevus syndrome: MR of intracranial involvement. AB - Focal thickening of the calvarium, hypoplasia of the white matter, cortical calcifications, and a leptomeningeal drape that enhanced after contrast injection were demonstrated by MR in the parietooccipital region ipsilateral to a facial sebaceous nevus in a patient with epidermal nevus syndrome. PMID- 8237714 TI - Sydenham chorea. PMID- 8237715 TI - More on handgun control. PMID- 8237716 TI - Cefadroxil reduces the production of IgE in a 3 year old asthmatic with juvenile rheumatoid arthritis. AB - Cefadroxil, a cephalosporin, had been prescribed to children with superinfected atopic dermatitis, and was shown to improve both the infection, the clinical status and induced a dramatic lowering of the serum total IgE levels. Further studies have confirmed the IgE immunomodulating properties of cefadroxil. We report the case of a 3 year old asthmatic child who was hospitalized for superimposed pneumonia and was included in a study evaluating cefadroxil. The child was also suffering of juvenile rheumatoid arthritis. After treatment with cefadroxil and oral salbutamol, the child fully recovered. The initially elevated serum IgE (day 1:556 IU/ml) dropped to normal values (day 21: 52 IU/ml), while the production of IgE in vitro by peripheral blood B cells was normalized. We suggest that one mechanism of action of cefadroxil is the stimulation of production of gamma interferon in patients with atopic disorders; this mechanism interferes with the IL-4 primary signal, as well as with other second signals recognized for the synthesis of IgE. Gamma interferon may also prove beneficial for the control of juvenile rheumatoid arthritis. PMID- 8237717 TI - In vitro donor-specific hyporesponsiveness and T cell subsets in renal allograft recipients. AB - In order to assess the immune mechanisms triggered by an immunosuppressive regimen consisting of prophylactic antilymphocyte globulin plus low-dose cyclosporine A and steroids, we studied the short-term evolution of both, anti donor in vitro alloresponse and peripheral blood T cell subsets in 21 recipients of a cadaveric kidney allograft. Spleen cells from cadaveric donors and peripheral blood lymphocytes from the respective recipients pretransplant (pre Tx), at three and six months posttransplant (post-Tx) were obtained to perform one-way mixed lymphocyte cultures and flow cytometry analysis of lymphocyte subsets. The results indicated the development of donor-specific mixed lymphocyte culture (MLC) hyporesponsiveness as early as three months post-Tx, paralleled by a decrease in CD4+CD29+ helper-inducer cells and by an increase in CD8+CD45RA+ suppressor lymphocytes in peripheral blood. These changes were reflected in a very good clinical outcome of the patients. The present results further suggest that suppression of the immune system just before transplantation is a suitable method to induce early specific hyporesponsiveness to the allograft. PMID- 8237718 TI - Specific IgE antibody response to human insulin in diabetic subjects. AB - Treatment with heterologous insulin preparations induce insulin antibody formation in most of the patients. A hundred of diabetic individuals in treatment with human insulin (HI) for at least 6 months at the time of the study and without any clinical immunological reaction, were selected in order to study the development of specific IgE antibodies against HI by in vivo and in vitro methods. Serologic specific IgE antibodies to HI were measured by RAST (Pharmacia) and 5 RAST-positive subjects were found. The specificity of these results were confirmed in 4 of the 5 subjects by RAST-inhibition. Thus, the prevalence of specific serologic IgE to HI in our study was 4% (1.1%-9.9%, p < 0.05). Only two of the RAST-positive subjects had cutaneous reactivity to HI by intradermal technique and one of the RAST-negative subjects presented also insulin skin reactivity. HI skin reactivity was found in 3% (0.6%-8.5%, p < 0.05) of the 100 diabetic subjects. These results differ from others obtained with heterologous insulins. PMID- 8237719 TI - Urticaria by Blastocystis hominis. Successful treatment with paromomycin. AB - Urticaria and angioedema are easily recognized disorders, but in at least 70 percent of individuals, chronic episodes of urticaria are of unknown causes. We present 10 cases of chronic urticaria associated parasitation by blastocystis hominis. This parasite has not been previously related with urticaria. Both intestinal parasitation as well as urticaria responded successfully to paromomycin sulfate. PMID- 8237720 TI - Cost-effectiveness of various methods of diagnosing hypersensitivity to Alternaria. AB - This study was undertaken for two reasons: 1) It is more difficult to diagnose hypersensitivity to molds than to other allergens, so an evaluation of diagnostic tests was needed. 2) Alternaria is the principal cause of mold sensitization in our area. Sixty-six patients (20 +/- 4 years) were selected and divided into two groups. Group A was made up of patients with rhinitis and/or asthma due to Alternaria sensitization. Group B consisted of patients sensitized to other allergens and patients with nonrespiratory allergic disorders. Skin tests (prick and intradermal), challenge tests (conjunctival, nasal, and bronchial), and specific IgE determination were performed for all patients. A biologically standardized extract of Alternaria tenuis (Alergia e Inmunologia Abello, S. A., Madrid, Spain) obtained from a single batch was used for all tests. Our diagnostic criterion was a clinical history of rhinitis or asthma that coincided with the results of nasal/bronchial challenge. The diagnostic value of the other tests was compared to this criterion. In the group of rhinitic patients, skin tests and conjunctival challenge were more sensitive than specific IgE determination. In asthmatic patients, the most sensitive techniques were nasal and conjunctival challenges, followed by prick and intradermal skin tests, and, lastly, serum specific IgE determination. When rhinitis and asthma were considered jointly, the most sensitive test was conjunctival challenge, followed by skin-prick and intradermal tests. All tests had the same specificity, regardless of disorder. Nasal challenge was positive in all patients. Skin tests are easy to perform, cheap, non-traumatic for the patient, and sufficiently specific and sensitive for the diagnosis of Alternaria hypersensitivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237721 TI - Plantain allergy (Plantago lanceolata): assessment of diagnostic tests. AB - A study was made of 55 patients with skin prick test results for Plantago lanceolata of 3 mm2 or more. They were evaluated in relation to the response elicited by 10 mg/ml of histamine HCl. Patients with a response to Plantago less intense than the response to histamine were considered as having a "negative" response, whereas those whose response to Plantago was more intense than the response to histamine were considered as having a "positive" response. In 35 cases a radioallergosorbent test (RAST) was performed and in 22, nasal provocative testing (NPT). The frequency of positive response was 0.63 for skin tests, 0.11 for RAST and 0.72 for NPT. Among the patients with positive NPT, the frequencies of positive response to skin tests and RAST were 0.71 and 0.6 (p < 0.05), respectively. Results are discussed and study methods are proposed. PMID- 8237722 TI - Retrospective histopathology of endocrine pancreas in bronchial asthma. AB - A retrospective histopathological, immunocytochemical and morphometric study was done on samples of pancreas of 16 asthmatic dead humans in comparison with 6 dead control subjects, by other causes not related with allergic or endocrine diseases. A ribbon-like type of islet was the only histological feature found with the exception of one case of chronic pancreatitis. A significative increase of insulin islets (p < 0.05) was observed in the asthmatics patients while no differences in calculated area, perimeter and diameter of pancreatic islets were recorded. A probable hyperplasia of beta-cells could be related to the onset of the asthmatic crisis based on the pro-inflammatory stimulation of insulin or the action of anti-asthmatic treatments in the acute attack. PMID- 8237723 TI - Confronting violence: role of the family physician. PMID- 8237724 TI - Carotid endarterectomy: to do or not to do. PMID- 8237725 TI - Naproxen and the occurrence of eosinophilic pneumonia. PMID- 8237726 TI - Akathisia in association with nortriptyline therapy. PMID- 8237727 TI - Family practice research at the National Institutes of Health. PMID- 8237728 TI - Pulmonary arteriovenous malformation in pregnancy. PMID- 8237729 TI - Symptomatic carotid artery disease: current management recommendations. AB - Proper management of symptomatic carotid artery disease requires prompt diagnosis and therapy based on both the patient's symptoms and the nature of the carotid lesion. Duplex ultrasonography is the preferred diagnostic modality for evaluating symptomatic patients for the presence of a hemodynamically significant carotid lesion. Arteriography can confirm severe carotid stenosis or delineate a nonstenotic, ulcerated plaque before surgery. Antiplatelet and anticoagulant agents administered after transient ischemic attacks or completed stroke have shown questionable benefit in stroke reduction as an independent variable. Results of randomized clinical trials support the use of carotid endarterectomy for symptomatic patients with ipsilateral carotid stenosis greater than 70 percent. The operation should be performed for appropriate indications by surgeons whose perioperative morbidity and mortality rates meet established guidelines. PMID- 8237730 TI - Perfusion imaging in ischemic heart disease. AB - Myocardial perfusion imaging with thallium-201 has been used extensively for many years in the evaluation of ischemic heart disease. It is a valuable adjunct to exercise treadmill testing and is especially helpful in evaluating cardiac perfusion in patients who have abnormal resting electrocardiograms, who are unable to exercise, and whose clinical history and exercise test results are discordant. Perfusion imaging is useful for both diagnosis and prognosis in patients with coronary artery disease. In patients unable to exercise, the coronary vasodilators dipyridamole and adenosine may be used in conjunction with thallium imaging. Two recently approved technetium agents, sestamibi and teboroxime, may be used in place of thallium. PMID- 8237731 TI - Oral hypoglycemic agents in the treatment of type II diabetes. AB - The cornerstones of therapy for non-insulin-dependent diabetes mellitus are patient education, dietary modification with weight loss, and regular exercise. If diet and exercise fail to control hyperglycemia, pharmacologic intervention is warranted. The sulfonylureas are the only hypoglycemic agents approved for use in the United States. Their hypoglycemic effect is achieved through stimulation of insulin secretion and reduction of insulin resistance in the peripheral tissues. Two classes of sulfonylureas are available. First-generation drugs, such as tolbutamide and tolazamide, are less expensive and less powerful than second generation drugs, such as glipizide and glyburide. PMID- 8237732 TI - Helping patients choose appropriate contraception. AB - Helping patients choose the proper contraceptive requires familiarity with the specific advantages and disadvantages, as well as failure rates, of available methods. Spermicides, condoms and contraceptive sponges are inexpensive and do not require physician prescription. Diaphragms and cervical caps help protect against sexually transmitted diseases but are more difficult to use effectively. Oral contraceptives are highly effective but have estrogen- and progesterone related side effects. Norplant is ideally suited for less motivated users but requires an office surgical procedure. Depo-Provera provides another long-term option but may result in amenorrhea. Intrauterine devices provide long-term contraception without hormonal side effects but increase the risk of pelvic infection and may cause heavy menstrual bleeding. Tubal ligation and vasectomy are options for permanent contraception when childbearing is no longer desired. PMID- 8237733 TI - AAP policy statement on screening, treatment and prevention of lead poisoning. PMID- 8237734 TI - Neurocysticercosis in the southwestern United States. PMID- 8237735 TI - The physician's role in preventing elder abuse. PMID- 8237736 TI - Hearing screening in kindergarteners. PMID- 8237737 TI - Use of mesalamine and olsalazine in ulcerative colitis. PMID- 8237738 TI - The second revolution in family practice has begun. PMID- 8237739 TI - Diary from a week in practice. PMID- 8237740 TI - Complications of vasectomy. AB - Family physicians should be aware of the potential effects and complications of vasectomy so they can appropriately counsel patients seeking sterilization. Vasectomy produces anatomic, hormonal and immunologic changes and, although not substantiated by clinical studies, has been reputed to be associated with atherosclerosis, prostate cancer, testicular cancer and urolithiasis. Complications of vasectomy include overt failure, occasional sperm in the ejaculate, hematoma, bleeding, infection, sperm granuloma, congestive epididymitis, antisperm antibody formation and psychogenic impotence. Compared with tubal ligation, vasectomy has fewer serious complications and a comparable failure rate. PMID- 8237741 TI - Basal cell carcinoma. AB - Fair-skinned persons who have had significant exposure to the sun are at greatest risk for basal cell carcinoma. Clinically, basal cell carcinomas include numerous nodular and flat types. The lesions of basal cell carcinoma are usually asymptomatic until enlargement, bleeding or invasion of underlying tissue occurs. Metastasis is rare. A biopsy should be obtained before destructive treatment is carried out. Surgical excision, curettage and electrodesiccation or cryosurgery are acceptable procedures for removal of most lesions. Periodic follow-up is necessary to screen for new or recurrent skin cancer. PMID- 8237742 TI - Treatment of nausea and vomiting in pregnancy. AB - The nausea and vomiting that often accompany early pregnancy may be ameliorated if the patient eats small portions of food at frequent intervals, increases the amount of carbohydrates and decreases the amount of fat, and avoids bothersome food odors. When these strategies fail, antiemetic drugs such as pyridoxine and phosphorated carbohydrate solution may help, as well as doxylamine, promethazine or trimethobenzamide. When drugs are considered for use in pregnant patients, the safety of the patient and the fetus must always come first. PMID- 8237743 TI - APA issues practice guideline for major depressive disorder in adults. American Psychiatric Association. PMID- 8237744 TI - AMA guidelines on HIV infection. PMID- 8237745 TI - Effects of stent design and serum cholesterol level on the restenosis rate in atherosclerotic rabbits. AB - We investigated the effect of serum cholesterol level and stent design on the restenosis rate within the stent after balloon angioplasty and stent implantation using atherosclerotic rabbits. Two types of nickel/titanium stents with gaps (open stent) and without gaps (closed stent) between the wire coils were implanted into the aorta of the rabbits 10 weeks after atherosclerosis had been induced using a standard high cholesterol diet and balloon abrasion. Each rabbit had an open stent and a closed stent implanted into the infrarenal abdominal aorta. Between these two stents a control segment of the aorta was treated with angioplasty alone. The animals were divided into two groups according to the diet protocol as follows: in group I (n = 9) a high cholesterol diet was stopped after stent implantation; in group II (n = 10) a high cholesterol diet was maintained after stent implantation. Digital subtraction angiograms were obtained every 4 weeks for up to 24 weeks and the narrowest diameter of the arterial segments within each stent and in the segment between stents was measured. The diameter narrowing within the closed stent was greater in the high cholesterol group compared with the low cholesterol group: 12 weeks (2.57 +/- 0.09 mm in group I vs 2.14 +/- 0.15 mm in group II, mean +/- S.E., p < 0.05); 16 weeks (2.55 +/- 0.09 mm vs 2.14 +/- 0.12 mm, p < 0.05); 20 weeks (2.59 +/- 0.06 mm vs 1.98 +/- 0.12 mm, p < 0.01); and 24 weeks (2.45 +/- 0.11 mm vs 2.01 +/- 0.11 mm, p < 0.05). No significant differences in the narrowest diameter of the arterial segments were observed between high and low cholesterol groups in the angioplasty alone areas or within the open stents. There was a significant difference in the narrowest diameter between stents with versus those without gaps (at 12, 16, and 20 weeks poststenting in group I and at 4, 8, 12, 16, 20, and 24 weeks in group II). Thus the stent with the least metal is correlated with less stenosis and intimal hyperplasia. From these data we conclude that both stent design and serum cholesterol are important factors for restenosis after stent implantation. PMID- 8237746 TI - Subcutaneous heparin in postangioplasty management: comparative trial with intravenous heparin. AB - Abrupt closure of a coronary artery after successful angioplasty remains a problem for the interventionalist. Many laboratories continue to administer heparin intravenously for 12 to 24 hours in an attempt to alleviate this problem. But prolonged heparin therapy delays sheath removal and may lead to groin and vascular complications, and so prolong the hospital stay. To test the hypothesis that subcutaneous heparin was as efficacious as intravenous heparin in preventing acute closure while reducing the vascular complications associated with extended sheath placement, we prospectively randomized 151 patients to two groups. The intravenous group, 77 patients, received continuous intravenous heparin at 1000 units/hour for 12 to 18 hours; the subcutaneous group, 74 patients, received 12,500 units subcutaneously every 12 hours for three doses after sheath removal 2 to 3 hours after the angioplasty. The activated clotting time immediately after the angioplasty was 401 +/- 108 seconds in subcutaneous group, as compared with 368 +/- 67 seconds in the intravenous group (p = 0.028). Patients receiving subcutaneous heparin continued to show adequate anticoagulation, with a PTT of 85 +/- 21 seconds obtained approximately 12 hours after the procedure. The PTT at discharge was statistically greater in the subcutaneous group, at 49.2 +/- 21 seconds, versus 35.6 +/- 13 seconds in the intravenous group (p < 0.001). Abrupt occlusion was similar in both groups, but the hematomas and bleeding/oozing in the intravenous group was significantly higher when compared with that of the subcutaneous group, 16 versus 6 (p = 0.026) and 26 versus 7 (p < or = 0.002), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237747 TI - Short- and long-term neurohormonal activation following acute myocardial infarction. AB - Plasma levels of atrial natriuretic peptide (ANP), angiotensin II, aldosterone, and catecholamines were followed for 1 month and then for 4 to 6 months in 55 patients with acute myocardial infarction. Plasma hormones were highest within the first 24 hours after the onset of infarction but normalized during the first few days in patients without heart failure. In patients with symptoms of heart failure, angiotensin II and norepinephrine remained elevated for 1 month and ANP remained elevated for 4 to 6 months. During head-up tilt, angiotensin II and norepinephrine increased most in patients with overt heart failure. Among patients with a first myocardial infarction, a positive correlation was found between infarct size and ANP, angiotensin II, and norepinephrine on day 5 to 7 and between infarct size and angiotensin II during head-up tilt at 1 month, and between infarct size and ANP at 1 month. A similar relationship was found when only patients without heart failure were studied. It is concluded that sustained neurohormonal activation after myocardial infarction mainly occurs in patients with clinical heart failure but is related to the magnitude of myocardial damage, even in patients without heart failure. Measurement of neurohormones during head up tilt may be an additive assessment for the detection of neurohormonal activation. PMID- 8237748 TI - Pretreatment with theophylline does not affect adenosine-induced thallium-201 myocardial imaging. AB - This study was designed to evaluate the effects of intravenous theophylline given before intravenous adenosine for thallium-201 imaging. Sixteen patients with known reversible thallium-201 defects were randomized to a double-blind crossover study of a 45-minute infusion of placebo or theophylline (given as the ethylenediamine salt, aminophylline, mean concentration 16.1 mg/L) before adenosine thallium-201 imaging. Adenosine was infused for 6 minutes at a rate of 140 micrograms/kg/min. Thallium-201 (2.5 to 3.0 mCi) was administered after 3 minutes of infusion. Blood pressure, heart rate, symptoms, and the electrocardiogram were monitored continuously. Planar thallium-201 imaging was obtained in three standard views and was interpreted using blinded segmental analysis and computerized quantitation. Systolic and diastolic blood pressure at baseline and during adenosine administration were similar following treatment with theophylline and placebo. The increase in heart rate observed during adenosine infusion was significantly reduced by theophylline pretreatment. Adenosine-induced symptoms (both cardiac and noncardiac) as well as ischemic electrocardiographic changes were significantly reduced after theophylline infusion (p < 0.05). In one patient, Mobitz type II heart block seen during adenosine infusion following placebo was absent with theophylline pretreatment. The size of adenosine-induced thallium-201 defects was unchanged by theophylline infusion using either segmental analysis (8 +/- 4 vs 9 +/- 5) or a computerized score (47 +/- 27 vs 45 +/- 21). Despite reduction in both symptoms and ischemic electrocardiographic changes, theophylline does not alter thallium-201 imaging following intravenous adenosine infusion. PMID- 8237749 TI - Prolonged bradyarrhythmias after isolated coronary artery bypass graft surgery. AB - To evaluate clinical and electrocardiographic (ECG) characteristics that may predict the occurrence of bradyarrhythmias after isolated coronary artery bypass graft (CABG) surgery, 1614 consecutive patients who had this procedure performed at our institution from January 1988 to December 1990 were reviewed. Thirteen (0.8%, 7 males and 6 females) patients had prolonged (mean 10.5 +/- 6.5 days) postoperative bradyarrhythmias and required insertion of a permanent pacemaker. Complete heart block occurred in eight patients and sinus node dysfunction in five. These 13 patients (group A) were compared with a group of 490 arbitrarily selected CABG patients (group B) without bradyarrhythmias whose preoperative ECGs were reviewed. Patients in group A were older (mean 69.2 vs 62.8 years; p = 0.0004) and had concomitant left ventricular (LV) aneurysmectomy more frequently (p = 0.02) and internal mammary graft revascularization less frequently (p = 0.022) than group B patients. Review of preoperative ECGs revealed a higher occurrence of complete left bundle branch block (LBBB) (5 of 13 vs 6 of 490; p < 0.0001) and a borderline, more leftward frontal plane QRS axis (-5.3 vs 13.1 degrees, p = 0.068) in group A patients. There were no differences between the groups with respect to gender, number of bypass grafts, location of prior myocardial infarction, and preoperative ECG intervals (PR, QRS, QTc). Multivariate analysis identified the presence of a preoperative LBBB, concomitant LV aneurysmectomy and age > 64 years as independent predictors of severe and prolonged postoperative bradyarrhythmias, mainly complete heart block.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237750 TI - Coronary artery size in mitral regurgitation and its regression after mitral valve surgery. AB - The relationship between coronary artery size and left ventricular (LV) muscle mass was studied in 10 control subjects and in 10 patients with chronic mitral regurgitation before and 28 +/- 15 months after mitral valve surgery. Left and right coronary artery size was determined by quantitative coronary arteriography. Left coronary artery size was significantly increased before surgery (26 mm2) and decreased after operation (23 mm2), but was still larger than in control subjects (14 mm2). The right coronary artery was also enlarged preoperatively (13 mm2; controls = 9 mm2), but was normalized after surgery (11 mm2). A linear correlation was found between LV muscle mass and left (r = 0.88, p < 0.001) and right coronary artery size (r = 0.84, p < 0.001) as well as between right coronary artery size and mean pulmonary artery pressure (r = 0.56, p < 0.01). Thus in chronic mitral regurgitation the enlargement of the left and right coronary artery is proportional to the degree of LV hypertrophy. The increase in right coronary artery size is probably the result of right ventricular pressure overload. Postoperatively there is only partial regression of left coronary artery size but normalization of right coronary artery size. PMID- 8237751 TI - Comparative prognostic value of clinical risk indexes, resting two-dimensional echocardiography, and dipyridamole stress thallium-201 myocardial imaging for perioperative cardiac events in major nonvascular surgery patients. AB - The relative prognostic value of widely accessible resting two-dimensional echocardiographic ventricular function data has not been compared with recognized clinical and scintigraphic risk markers in patients who are unable to exercise before major nonvascular surgery. To this end, 53 consecutive patients aged 67 +/ 13 years undergoing preoperative evaluation (intraabdominal, 23%; orthopedic, 30%; thoracic, 9%; other, 38%) for known or suspected coronary artery disease were followed up to evaluate the prognostic value of these studies for the perioperative cardiac events (cardiac death [n = 4], myocardial infarction [n = 2], unstable angina [n = 3], and pulmonary edema [n = 8]) that occurred in 13 of the 53 patients (25%). Dipyridamole thallium-201 myocardial redistribution defects occurred in 15 (28%) patients. Resting echocardiographic left ventricular dysfunction was present in 21 (40%) patients. Multivariate analysis of clinical, echocardiographic, and scintigraphic risk predictors revealed that cardiac events were not predicted by clinical variables, including Goldman class or score. Cardiac events were independently predicted only by the presence of significant left ventricular dysfunction on resting two-dimensional echocardiography (p < 0.042) and dipyridamole thallium-201 defect redistribution (p < 0.026). A dipyridamole-induced reversible thallium-201 perfusion defect was predictive of subsequent cardiac death or myocardial infarction (p < 0.02), whereas left ventricular dysfunction on resting echocardiography was predictive of perioperative pulmonary edema (p < 0.023). We conclude that stress thallium-201 perfusion imaging and resting two-dimensional echocardiography provide independent prognostic information in patients undergoing major nonvascular surgery who are at significant risk for ischemic cardiac events and who are unable to perform standard exercise stress tests.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237752 TI - Effects of canine myocardial infarction on sympathetic efferent neuronal function: scintigraphic and electrophysiologic correlates. AB - We studied the effects of nondecentralized left stellate ganglion stimulation on regional epicardial monophasic action potential duration at 50% (APD50) and 90% (APD90) repolarization from 104 sites in 10 surviving dogs with a chronic myocardial infarction model. These effects were correlated with thallium-201 and iodine-123 metiodobenzylguanidine (MIBG) imaging to identify areas of viable but denervated myocardium. Mean infarct size was 5.2% +/- 0.8% total heart weight, and the planimetered areas of denervation were always larger (18% +/- 4% total heart area). During constant ventricular pacing, stellate stimulation tended to shorten the APD90 only in normally innervated areas (364 +/- 5 to 358 +/- 5 msec) and to increase in denervated areas (358 +/- 5 to 362 +/- 5 msec), (p value not significant (NS) for prestellate and poststellate stimulation; p < 0.05 for difference between denervated vs innervated). The APD50 significantly shortened in innervated areas from 287 +/- 5 to 270 +/- 3 msec (p < 0.05) compared with denervated areas (283 +/- 4 to 274 +/- 5 msec, p = NS). We conclude that MIBG imaging demonstration of denervation identifies areas with impaired shortening of the epicardial APD50 in response to stellate stimulation and that nontransmural myocardial infarction produces areas of denervation larger than areas of necrosis. PMID- 8237753 TI - Determinants of inducible ventricular tachycardia in patients with clinical ventricular tachyarrhythmia and no apparent structural heart disease. AB - Thirty-seven patients with symptomatic ventricular tachyarrhythmia and no apparent structural heart disease were evaluated with multiple cardiovascular tests to establish the relationship between the results of programmed electric stimulation and other clinical and arrhythmia variables. Of 37 patients, 12 (32%) had inducible sustained ventricular tachycardia. Factors associated with the results of programmed electric stimulation included a history of ventricular tachycardia > or = 30 seconds requiring intervention for termination and global right heart abnormality documented by echocardiography. During treatment for a mean follow-up of 50 months, 29 patients did well, 6 patients had recurrences of major arrhythmia symptoms, 1 was lost to follow-up and 1 had a noncardiac death. Those patients with an adverse outcome were more likely to have inducible ventricular tachycardia. Thus certain clinical and echocardiographic data are associated with the results of programmed electric stimulation, which in turn have important prognostic value in this group of patients. Sustained ventricular tachycardia is unlikely to be induced in patients with no evidence of structural heart disease and clinical nonsustained ventricular tachycardia. PMID- 8237754 TI - Evidence for a reexcitability gap in man after treatment with type I antiarrhythmic drugs. AB - The intention of this study was to determine whether type IA antiarrhythmic drugs cause a disparity between refractoriness and repolarization, and if so, its magnitude. We simultaneously measured monophasic action potential duration (MAPD) and effective refractory period (ERP) at a right ventricular site in 11 patients without overt right ventricular disease. The pacing protocol, which included sinus rhythm and a 600 and 400 msec cycle length of ventricular drive, was performed at baseline and after the intravenous administration of 15 mg/kg of procainamide in nine patients, and of 10 mg/kg of quinidine in two patients. Despite trivial changes in sinus rates, drug therapy caused a 10% to 17% increase in MAPD and ERP that shortened with decreasing drive cycle lengths. At baseline there was a small gap (10 to 13 msec) between MAPD and ERP in sinus rhythm and with a 600 or 400 msec drive. However, the type IA drug caused a significant widening of this gap that was most profound in sinus rhythm (45 msec) and that shortened but was not abolished with a 600 and 400 msec drive (28 and 29 msec, respectively). The disparity was caused in one third of cases by postrepolarization refractoriness. Type I drugs increase the difference between repolarization and refractoriness, and this effect is partially reversed with increases in heart rate. The clinical implications of these findings are discussed. PMID- 8237755 TI - Verapamil for the suppression of idiopathic ventricular tachycardia of left bundle branch block-like morphology. AB - This study examines the efficacy of verapamil for the suppression of idiopathic ventricular tachycardia (VT) of left bundle branch block LBBB-like morphology. Forty-two patients (mean age 36.2 +/- 12.1 years; 20 men and 22 women) with VT and without any underlying cardiac abnormality on clinical examination and noninvasive investigation were studied. The inducibility of clinical VT was examined by treadmill exercise testing and programmed ventricular stimulation (PVS). In 29 patients VT was inducible by exercise testing, in 24 by PVS, and in 23 there was evidence of VT on Holter monitoring. After baseline testing, patients were treated with verapamil 120 mg thrice daily for at least 5 half lives for the drug to load before evaluation. With Holter monitoring, 74% of patients with evidence of VT at baseline testing demonstrated a change of status from nonsustained VT to no VT or from sustained VT to nonsustained VT. Four patients had nonsustained VT during verapamil treatment but no VT at baseline. There was a significant reduction in the number of ventricular ectopic beats over 24 hours (baseline: 15,541 +/- 17,599 vs verapamil treatment: 8892 +/- 15,582, p < 0.01). Exercise-induced VT was suppressed in 56% of patients with VT during baseline testing, but no effect of verapamil on the tachycardia was observed in 26%. The remaining patients demonstrated a partial response to verapamil; the rate of VT was unchanged, although the duration of the runs was reduced. Sustained monomorphic VT was inducible in only 5 patients, of whom 4 were rendered noninducible; 1 patient remained inducible.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237756 TI - Characteristics of Ca(2+)-activated K+ channels isolated from the left ventricle of a patient with idiopathic long QT syndrome. AB - Early afterdepolarizations (EADs), possibly caused by reduced K+ conductance, have been hypothesized to cause the long QTU interval and ventricular tachyarrhythmias (VT) in patients with the long QT syndrome (LQTS). In a 26-year old woman with aborted sudden death as a consequence of the idiopathic LQTS, we recorded with a contact electrode left ventricular endocardial EADs that were enhanced by epinephrine and phenylephrine. Because of uncertain efficacy and side effects achieved with beta-adrenoceptor blockade, the patient underwent left sided cardiac sympathectomy, at which time we obtained left ventricular biopsy tissue. Crude membrane vesicles were prepared from this tissue and single-channel activity was studied after incorporation of the vesicles in an artificial lipid bilayer (phosphatidylserine, phosphatidylethanolamine, 4:5 weight ratio in decane) in the tip of a patch clamp pipette. Bath and pipette contained 100 mmol/L KCI and 25 mmol/L N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) at pH 7.4. We recorded K+ conducting channels with a mean slope conductance of 49.9 +/- 4.7 picosiemens (pS) (n = 5). Channel open probability was increased by the addition of 1 to 10 mumol/L Ca2+ to the experimental chamber. Addition of charybdotoxin (1-3 nmol/L), a known specific inhibitor of Ca(2+)-activated K+ channels, blocked channel activity. These results are the first to demonstrate Ca(2+)-activated K+ channels from a patient with idiopathic LQTS. These channels appear to show normal characteristics when studied in an artificial planar lipid bilayer. PMID- 8237757 TI - Invasive electrophysiologic evaluation in octogenarians: is age a limiting factor? AB - To assess the indications, diagnostic yield, and incidence of complications of electrophysiologic testing in the elderly we reviewed our experience with 60 procedures in 45 patients aged > or = 80 years (range 80 to 92 years, mean age 83) undergoing full electrophysiologic evaluation in our laboratory over the past 7 years. The yield of inducible ventricular tachycardia (31%), supraventricular tachycardia (4%), and previously unsuspected conduction abnormalities significant enough to warrant permanent pacemaker implantation (9%), together with the low incidence of complications (1 patient had a deep venous thrombosis and femoral artery pseudoaneurysm, representing an incidence of 2.2% of patients undergoing studies or 3.3% incidence of complications per procedure), suggest that invasive electrophysiologic procedures in the elderly can provide useful information at a complication rate comparable with that of younger patients. PMID- 8237758 TI - Percutaneous balloon mitral valvuloplasty: analysis of echocardiographic and other variables related to outcome. AB - To determine whether mitral valve (MV) morphology influences the result of balloon mitral valvuloplasty (BMV) for mitral stenosis, two-dimensional echocardiography was performed before BMV in 53 patients and in 25 normal controls. The two-dimensional echocardiographic features of MV leaflets: thickness, length and motion, diastolic MV excursion, chordal length, MV annular diameter (MVAnD), subvalvular distance ratio (SDR), and effective balloon dilating area (EBDA) and diameter (EBDD) were then correlated to the immediate post-BMV mitral valve area (MVA). For the total patient population, post-BMV MVA increased from 0.76 +/- 0.24 to 1.91 +/- 0.59 cm2 (p < 0.0001) and mean diastolic transmitral gradient decreased from 20.1 +/- 6.15 to 5.8 +/- 3.29 mm (p < 0.0001). The patients were divided into two groups on the basis of post-BMV MVA. Group I had post-BMV MVA < 2.0 cm2 and group II had post-BMV MVA > or = 2.0 cm2. A statistically significant difference was noted in SDR (0.33 +/- 0.057 vs 0.45 +/- 0.042, p < 0.0001); mid-MV anulus to tip of papillary muscle (PM) distance (20.0 +/- 3.8 vs 27.9 +/- 4.54 mm, p < 0.0001); chordal length (4.3 +/- 3.6 vs 9.8 +/- 3.9 mm, p < 0.0001); diastolic MV excursion (15.5 +/- 2.6 vs 18.2 +/- 4.2 mm, p < 0.01); leaflet mobility (p < 0.05); and EBDA (4.4 +/- 0.6 vs 4.9 +/- 0.5 cm2, p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237759 TI - Long-term results after balloon pulmonary valvuloplasty in adults. AB - Percutaneous balloon pulmonary valvuloplasty for congenital pulmonary valve stenosis was performed in 40 adult patients (aged 18 to 56 years). The duration of follow-up was 24.5 +/- 11.5 months. The peak systolic pressure gradient from the right ventricle to the pulmonary artery decreased from 107 +/- 29 mm Hg to 37 +/- 25 mm Hg immediately after the procedure. On follow-up, gradient obtained either by cardiac catheterization or Doppler echocardiography was 31 +/- 13 mm Hg. In eight patients with residual gradient after valvuloplasty of > 50 mm, the mean peak systolic gradient decreased from 74.5 +/- 18.3 to 33.5 +/- 13.9 mm Hg on follow-up. Thus balloon pulmonary valvuloplasty is a safe and effective procedure for the treatment of adult patients with pulmonic stenosis; there is a tendency for high residual gradients to regress on follow-up. PMID- 8237760 TI - Valve area and cardiac output in aortic stenosis: quantification by magnetic resonance velocity mapping. AB - Valve area and cardiac output were determined with magnetic resonance (MR) velocity mapping in 12 patients with aortic stenosis. Heart catheterization, Doppler echocardiography, and indicator dilution were performed for comparison. Left ventricle could be catheterized in only nine patients; in these cases, MR measured a mean valve area of 1.2 cm2 compared with 0.9 cm2 derived from catheterization data, with a mean difference of 0.2 cm2 between the 2 methods. The limits of agreement were [0.0, +0.5] cm2, less in patients with an important degree of concomitant regurgitation. In the whole material, MR measured a mean area of 1.1 cm2 compared with 1.2 cm2 derived from Doppler echocardiography data, with a mean difference of 0.1 cm2 and [-0.5, +0.6] cm2 as limits of agreement. In 11 patients the cardiac output was quantified by MR to a mean of 4.9 L/min and by indicator dilution to 5.0 L/min, with a mean difference of 0.2 L/min, and [-0.6, +0.8] L/min as limits of agreement. In addition, MR offers the major advance of simultaneous quantification of regurgitant volume in cases of concomitant regurgitation. In conclusion, because the two important prognostic determinants in aortic stenosis--the valvular area and the cardiac output--may be quantified, MR has potential to become a clinical tool in assessment of severity in aortic stenosis. PMID- 8237761 TI - Cine magnetic resonance imaging and color Doppler flow mapping displays of flow velocity, spatial acceleration, and jet formation: a comparative in vitro study. AB - To study the effects of flow acceleration and high-velocity jets on the display characteristics of cine magnetic resonance imaging compared with color Doppler flow mapping, a custom-designed in vitro flow model was developed. This model consisted of a funnel segment tapering to an orifice (0.78 cm2) that leads into a confined receiving chamber with a second, discrete orifice (0.78 cm2) at its distal end. Cine magnetic resonance images obtained at varying flow rates (1.5 to 27.2 L/min) demonstrated loss of signal intensity throughout the tapering zone of spatial acceleration and a small zone of more marked signal loss immediately proximal to the second orifice (always < 50% of the signal intensity within the tapering funnel zone) associated with more rapid spatial acceleration. A formed jet was imaged distal to the first orifice, and the turbulence area surrounding the laminar central jet core correlated well with flow rate (r = 0.98), as did the distance from the orifice to the subsequent onset of flow relaminarization (r = 0.96). A turbulent spray area was always seen distal to the second, discrete orifice. Comparative observations with color Doppler flow mapping and continuous wave Doppler demonstrated that signal intensity on cine magnetic resonance imaging is reduced by both spatial acceleration, and the high-velocity and turbulent jets associated with obstructive and regurgitant lesions. In vitro evaluation of cine magnetic resonance imaging allows comparative observations to be made about the flow characteristics of cine magnetic resonance imaging and color Doppler flow mapping and provides a more rational basis for the interpretation of cine magnetic resonance imaging in the clinical setting. PMID- 8237762 TI - Timing of recipient atrial contraction: a major determinant of transmitral diastolic flow in orthotopic cardiac transplantation. AB - The effect of recipient left atrial contraction on diastolic transmitral flow was studied in eight asymptomatic heart transplant patients (seven men and one woman, mean age 51.8 +/- 9.7 years) with normal left ventricular systolic function. The transmitral flow velocity profile was evaluated in 326 beats (mean 40.8 +/- 12 beats, range 20 to 58 beats/patient). Recipient left atrial contraction was classified as early diastolic, late diastolic, and systolic. Results showed: (1) early diastolic transmitral flow velocity integral (in centimeters) was lower (p < 0.05) following late diastolic recipient left atrial contractions (7.5 +/- 2.1) compared with early diastolic (10.1 +/- 2.7) or systolic (8.5 +/- 1.5) recipient left atrial contractions; (2) late diastolic transmitral flow velocity integral was higher following late diastolic (4.7 +/- 1.8) compared with systolic (4.1 +/- 1.9) recipient left atrial contractions and was virtually abolished (0.45 +/- 14) following early diastolic recipient left atrial contractions; and (3) total diastolic transmitral flow velocity integral was significantly higher (p < 0.05) following late diastolic and systolic (12.2 +/- 2.8 and 12.0 +/- 2.5) compared with early diastolic left atrial contractions (10.5 +/- 2.8). Thus recipient left atrial contraction is a significant determinant of the transmitral diastolic flow pattern in heart transplant patients. Variations in the timing of recipient left atrial contraction may lead to fluctuations of diastolic flow and may compromise cardiac output. PMID- 8237763 TI - Abnormal Valsalva blood pressure response in dilated cardiomyopathy: association with "pseudonormalization" of echocardiographic Doppler transmitral filling velocity pattern. AB - In patients with dilated cardiomyopathies, the presence of an abnormal blood pressure response to the Valsalva maneuver has been shown to correlate well with increased left-sided filling pressures. The presence of a "pseudonormalized" early peak to late peak velocity (E/A) ratio on echocardiographic Doppler transmitral filling pattern has also been correlated with increased left ventricular end-diastolic pressures in these patients. Since both abnormal blood pressure response to the Valsalva maneuver and a "pseudonormalized" Doppler transmitral E/A ratio probably represent elevated left ventricular filling pressure, we postulated that there would be a positive correlation between the two in a group of patients with dilated cardiomyopathy and abnormal systolic function. Twenty-five consecutive male patients with New York Heart Association (NYHA) class II to IV heart failure and dilated cardiomyopathy were included. Patients with abnormal blood pressure responses to Valsalva had significantly larger peak early (E) velocities, smaller peak late (A) velocities, and larger E/A ratios compared with patients with normal responses. E/A ratio < 1.0 was present in eight (100%) of the eight patients with a normal blood pressure response to the Valsalva maneuver and E/A ratio > 1.0 was present in 12 (71%) of 17 patients with an abnormal response (p < 0.01). This correlation supports the hypotheses of the physiologic mechanisms of these phenomena and also provides two noninvasive methods of evaluating left-sided filling pressures useful in the diagnosis and treatment of congestive heart failure caused by dilated cardiomyopathy. PMID- 8237764 TI - Right ventricular cardiomyopathy in identical and nonidentical young twins. AB - We describe the first sets of identical and nonidentical twins with right ventricular cardiomyopathy (RVC). Pair A: A 12-year-old boy was referred because of palpitation and syncope. Clinical and instrument examinations revealed an enlarged and depressed right ventricle (end-diastolic volume = 110 ml/m2; ejection fraction = 44%), spontaneous ventricular tachycardia, and fatty-fibrous infiltrates in the biopsy specimens. His asymptomatic, monozygotic twin showed localized involvement of the right ventricle with isolated, ventricular extrasystoles. Pair B: These 18-year-old nonidentical twin boys showed diffuse right ventricular involvement (end-diastolic volume = 110 ml/m2 and 114 ml/m2; ejection fraction = 30% and 24%, respectively), induction of sustained and nonsustained ventricular tachycardia, respectively, and fibrosis on endomyocardial biopsy. One of the boys died suddenly at rest after documented ventricular fibrillation. These cases support the hypothesis of a genetic etiology with a minor role for genotype and point to the important influence of environmental factors in determining the clinical features of the disease. PMID- 8237765 TI - Ventricular tachycardia in patients without structural heart disease. AB - The existence of ventricular tachycardia in subjects without detectable heart disease has been recognized for more than half a century. These rhythms occur in younger individuals and account for about 5% of referrals to the electrophysiology service. In contrast, ventricular tachycardia in the setting of chronic ischemic heart disease occurs in an older age group and accounts for more than 90% of all referrals. It is important to be able to distinguish between the different varieties of ventricular tachycardia, because in patients without structural heart disease sudden cardiac death is rare, compared with the very high mortality associated with postischemic recurrent ventricular tachycardia. The natural history of ventricular tachycardia occurring without structural heart disease tends to be benign even when the tachycardia is untreated. Thus these rhythms are more of an annoyance than a life-threatening event. This variety of ventricular tachycardia is frequently not inducible by atrial or ventricular programmed stimulation but is inducible by atrial or ventricular pacing, exercise testing, or isoproterenol infusion. The ventricular tachycardia in patients without structural heart disease is frequently responsive to drugs such as beta blockers, verapamil, or adenosine that have little or no effect on ventricular tachycardia associated with previous infarction. These differences strongly suggest the possibility of a tachycardia mechanism other than reentry, although conclusive evidence is still lacking. PMID- 8237766 TI - Simultaneous assessment of left ventricular perfusion and function with technetium-99m sestamibi after coronary artery bypass grafting. AB - This study examined the left ventricular perfusion and EF by using simultaneous SPECT and first-pass radionuclide angiography with technetium 99m sestamibi in 95 patients after uncomplicated coronary artery bypass grafting. The patients were divided into those with normal EF and no previous myocardial infarction before surgery (group 1, n = 57), and those with abnormal EF or infarction (group 2, n = 38). The SPECT images were normal in 37 patients in group 1 and in 6 patients in group 2 (p < 0.0001). The patients with normal SPECT images had a higher EF after surgery than those with abnormal images (65% +/- 10% vs 50% +/- 14%, p < 0.0001) and was higher in group 1 than in group 2 (64% +/- 8% vs 46% +/- 16%, p < 0.0001). There was a significant correlation between the EF and the extent of perfusion abnormality (r = -0.44, p < 0.0001). The patients with normal SPECT images could not be separated from those with abnormal images based on peak CK, CK-MB, and the electrocardiographic changes. Of the 69 patients with postoperative EF > or = 50%, the perfusion pattern was normal in 41 and abnormal in 28; of the 26 patients with EF < 50%, 24 had abnormal SPECT (p < 0.003). There was no significant change in mean EF after surgery (55% +/- 14% before vs 56% +/- 15% after). Thus simultaneous assessment of left ventricular perfusion and function after coronary artery bypass grafting showed that an abnormal perfusion pattern may exist despite a normal EF. These patients could not be predicted by enzymes or electrocardiographic changes. PMID- 8237767 TI - Angioscopy: a valuable tool in the deployment and evaluation of intracoronary stents. AB - Intracoronary stents were designed to improve balloon dilation results; however, to accomplish this, various criteria of optimal stent deployment must be met. Standard imaging techniques are neither sensitive nor specific enough for intraprocedural use in the verification of these implantation parameters. To assess the usefulness of angioscopy in the procedural and follow-up evaluations of Palmaz-Schatz and Strecker coronary stent deployment, 17 patients underwent angioscopy, 15 during stent placement and 2 during follow-up for stent restenosis. In the latter cases, thrombus formation was suspected; however, angioscopy showed tissue subtotally occluding the lumen without thrombus, so thrombolytic therapy was avoided. Similarly, among the 15 intraprocedural assessments, angioscopy disclosed intravascular thrombus unappreciated on angiography in two cases; another patient at high risk for intravascular thrombus was found not to have clot. In four patients, angioscopy disclosed residual narrowing in need of redilation at the Palmaz-Schatz stent articulation site. Thus in 9 (53%) of 17 stent patients, angioscopic findings either guided therapeutic selection or significantly modified the anticipated procedure. Angioscopy offers important information critical to the accurate placement and evaluation of intracoronary stents. PMID- 8237768 TI - Emergency portable percutaneous cardiopulmonary bypass for left ventricular free wall rupture from myocardial infarction. PMID- 8237769 TI - Radiofrequency ablation of posteroseptal accessory pathways in patients with abnormal coronary sinus. PMID- 8237770 TI - Radiofrequency catheter ablation of a posteroseptal accessory pathway within a coronary sinus diverticulum. PMID- 8237771 TI - Response of the QT-sensing, rate-adaptive ventricular pacemaker to mental stress. PMID- 8237772 TI - Submammary implantation of a cardioverter-defibrillator with nonthoracotomy lead system. PMID- 8237773 TI - Atrial septal aneurysm simulating a left atrial mass diagnosed by transesophageal echocardiography. PMID- 8237774 TI - Unilateral pulmonary edema related to pulmonary artery compression resulting from acute dissecting aortic aneurysm. PMID- 8237775 TI - Detection of major aortopulmonary collateral arteries by transesophageal echocardiography in pulmonary atresia with ventricular septal defect. PMID- 8237776 TI - Anatomically corrected malposition: report of six cases. PMID- 8237777 TI - Congenital fibrous left ventricular diverticulum. PMID- 8237778 TI - Right congenital pericardial defects. PMID- 8237779 TI - Therapeutic approach to unstable angina: nitroglycerin, heparin, and combined therapy. PMID- 8237780 TI - Restenosis revisited: insights provided by quantitative coronary angiography. AB - In this editorial, the problem of restenosis after coronary balloon angioplasty and other transluminal interventions is reviewed from the perspective of quantitative coronary angiography. The review is largely based on the experience of the Thoraxcentre in the application of quantitative angiography to the study of restenosis over the past decade, with incorporation and discussion of relevant and significant contributions from other groups. Current discrepancies in the angiographic definition of restenosis are highlighted and the use of percent diameter stenosis or MLD as the measurement parameter of choice is objectively addressed. Perspectives on the pathologic paradigm of restenosis are briefly reviewed as a basis from which to evaluate quantitative angiographic information provided by various studies. Particular attention is then paid, in chronologic fashion, to discussion and elaboration of insights to the restenosis process provided by quantitative angiographic studies, which have led to the introduction of some new methodological approaches to the comparison of short- and long-term angiographic luminal changes after various interventions. A word of caution on the potential pitfalls of quantitative angiographic studies is provided and counterbalanced with a discussion of clinical correlations of quantitative angiographic measurements. Finally, a proposal is made for the application of quantitative angiographic measurements to randomized clinical trials for the purpose of comparing new interventional devices. PMID- 8237782 TI - States take lead on health care reform. PMID- 8237781 TI - More look-alike and sound-alike errors. PMID- 8237783 TI - Understanding drug-induced febrile reactions. PMID- 8237784 TI - Nutrition support pharmacist helps physician practice. PMID- 8237785 TI - The "i" plan. PMID- 8237786 TI - Infant formula products. PMID- 8237787 TI - Oral anticoagulation monitoring in a community pharmacy. AB - Efforts by pharmacists to become more involved in patient care move the profession beyond the traditional role of dispenser. These initiatives increase the professionalism of the pharmacist and carry the potential for monetary rewards. Monitoring oral anticoagulation therapy is an area in which pharmacists can use specialized drug knowledge and advances in technology to fulfill a special patient need in the community setting. This article has outlined factors that should be considered in the development of an oral anticoagulation therapy monitoring program. Pharmacists can apply these principles to their individual practice environments to determine whether such a program is feasible, practical, and profitable. Pharmacists interested in this type of program must then prepare themselves for the increased demands of documentation, quality assurance, and patient counseling. By implementing an anticoagulation service, pharmacists can improve patient outcomes and extend their practices into a new frontier of pharmaceutical care. PMID- 8237789 TI - American Industrial Hygiene Association expanded position statement on extremely low frequency and magnetic fields. PMID- 8237788 TI - 36 inches of counter space. PMID- 8237790 TI - An in vitro comparison of the permeation of chemicals in vapor and liquid phase through pig skin. AB - This study used pig skin to compare vapor and liquid permeation of benzene, n butanol, and toluene in vitro. Vapors of radio-labeled chemicals were generated by passing purified air through two saturators in series containing the labeled chemical. The generated vapor was directed into the donor compartment of a modified liquid permeation cell. For liquid permeation experiments, neat chemicals were dosed directly on the surface of the skin. The variability of the generated concentrations for the vapor phase of each chemical ranged from 3-7%. The mean flux of the liquid chemicals was significantly higher than those of the vapor phase. There was no significant difference in the flux of the individual chemicals in the liquid phase. In the vapor phase test, the flux of toluene and benzene were not significantly different; however, for n-butanol the flux was significantly lower than the for either benzene or toluene. PMID- 8237791 TI - Evaluation of shipboard formation of a neurotoxicant (trimethylolpropane phosphate) from thermal decomposition of synthetic aircraft engine lubricant. AB - MIL-L-23699 lubricants that are composed principally of trimethylolpropane triheptanoate (TMP) and tricresyl phosphate (TCP) have been shown to form a neurotoxicant, trimethylolpropane phosphate (TMPP), during pyrolysis and/or combustion. Mechanistically, TMPP is thought to irreversibly inhibit the GABA mediated inhibitory response and thereby produce epileptiform clonic/tonic seizures with convulsions followed by death. Thermal decomposition of MIL-L-23699 lubricant produces TMPP under laboratory conditions, but this product has not been detected in the workplace following actual fires. This study has examined whether TMPP is produced during an actual shipboard fire by placing the synthetic lubricant in a fire environment aboard the ex-U.S.S. Shadwell, Mobile, Alabama. Both biological and chemical analyses were performed on the thermally decomposed lubricant to ensure detection of the neurotoxic material. Under the conditions of this study, the formation of TMPP during a shipboard fire was confirmed. The implications of this finding for safe management of post-fire cleanup are discussed. PMID- 8237792 TI - Paraoccupational exposures to lead and tin carried by electric-cable splicers. AB - To test the hypothesis that electric-cable splicers contaminate their homes with lead and tin, nine splicers were matched with nine of their neighbors. House dust samples were collected in two areas within each home: a laundry room/dirty clothes area, and a composite sample from other areas in the house. Samples were analyzed by energy dispersive X-ray fluorescence for lead and tin (tin is a tracer to the occupational source of lead). The difference in the geometric mean lead concentrations in the laundry areas between the splicers' and neighbors' homes (1021 ppm and 390 ppm) was statistically significant (p < 0.025). The difference in concentrations from the other areas of the house (585 ppm and 329 ppm) was also significant (p < 0.05). Tin concentrations in house dust were very different between the two groups (p < 0.0005), suggesting that electric-cable splicers were contaminating their homes with lead and tin from work. Recommendations are included to prevent paraoccupational lead exposures by eliminating the pathways into the home. Another recommendation suggests that blood-lead levels be screened in children under the age of seven who live with electric-cable splicers. PMID- 8237793 TI - Successful reduction of silica exposures at a sanitary ware pottery. AB - Researchers from the National Institute for Occupational Safety and Health (NIOSH) conducted a joint survey with the New Jersey Department of Health (NJDOH) to measure crystalline silica exposures and evaluate the adequacy of the existing control measures for reducing these exposures at a sanitary ware pottery. This survey found that 95% of the personal and area samples from the Slip House, Casting, Glaze Spray, and Glaze Preparation Departments exceeded the NIOSH Recommended Exposure Level (87% exceeded the Occupational Safety and Health Administration Permissible Exposure Level) for crystalline silica. Three years later, a follow-up survey found statistically significant reductions in respirable crystalline silica exposures in two of four plant departments, and statistically significant reductions in area concentrations in all four plant departments. These reductions were accomplished through a combination of automating and enclosing the batching system in the Slip House and by replacing the mold parting compound with a nonsilica material, altering the method of dry sweeping, cleaning of castings while damp, improving exhaust ventilation at the spray booths, and improved housekeeping. PMID- 8237794 TI - Interpretation of urine results used to assess chemical exposure with emphasis on creatinine adjustments: a review. AB - This paper reviews the process of elimination of creatinine (CRE), and the limitations presented when using it to express urine concentrations. This literature review leads to three conclusions: (1) CRE excretion is subject to wide fluctuations due to specific internal and external factors; (2) the use of CRE to correct chemical concentrations in urine will not necessarily improve the correlation to the exposure dose for all chemicals (it may, in fact, worsen the result); and (3) other means of expressing urine concentration may offer greater accuracy towards estimating individually absorbed dose. PMID- 8237795 TI - Determination of pentamidine isethionate in air. AB - A concern currently exists regarding the potential for exposure of health care workers to pentamidine isethionate, a drug used for prevention and treatment of Pneumocystis carinii pneumonia in immunocompromised patients, including those with human immunodeficiency virus infection. In order to evaluate worker exposures, a sampling and analytical method for pentamidine isethionate in air has been developed. This method involves sampling with a 37-mm PVC membrane filter at 1 to 2 L/min, recovery with 3 mL of 50:50 ethanol:water with 0.085% phosphoric acid and 0.04% tetramethylammonium chloride in an ultrasonic bath for 10 min, and analysis by high performance liquid chromatography with fluorescence detection. The limit of detection is about 18 ng per sample, and the lower limit of quantitation is 50 ng per sample. Recoveries of pentamidine isethionate from PVC filters were 0.76 to 0.91 at 50 to 8820-ng levels of fortification. Samples were stable on PVC filters during 27 days of storage at room temperature. Because patients who are treated with pentamidine isethionate are at increased risk of contracting tuberculosis (TB), safety precautions for handling samples contaminated with TB were included in the sampling and analytical method. PMID- 8237796 TI - Percutaneous transluminal coronary angioplasty catheters versus fluid-filled pressure monitoring guidewires for coronary pressure measurements and correlation with quantitative coronary angiography. AB - The functional significance of a coronary stenosis can be assessed by measuring the translesional pressure gradient. Thirty-four patients were studied in the setting of percutaneous transluminal coronary angioplasty (PTCA) to evaluate the clinical relevance of the pressure gradient measurements by means of a PTCA balloon catheter. Both before and after PTCA, the mean pressure gradient across the stenosis was measured by means of a newly developed, 0.015-inch pressure monitoring guidewire, first with only the wire across the stenosis (delta Pw, considered as the actual gradient), and second with the deflated balloon catheter advanced over the wire in the stenosis (delta Pb). Pressure gradients were correlated with quantitative coronary angiography of the stenotic segment. Before PTCA, mean delta Pb was larger than delta Pw (62 +/- 14 vs 30 +/- 20 mm Hg; p < 0.01). After PTCA, delta Pb remained systematically higher than delta Pw (23 +/- 14 vs 3 +/- 5 mm Hg; p < 0.01), despite a significant reduction of percent area stenosis from 84 +/- 9 to 46 +/- 17%, and an increase in minimal obstruction area from 0.98 +/- 0.48 to 3.49 +/- 1.32 mm2. A significant correlation was found between delta Pw and percent area stenosis (r2 = 0.66), with a marked increase after percent area stenosis reached 80%. The correlation between delta Pb and percent area stenosis was weaker (r2 = 0.53), the scatter of the data was larger, and the inflection point of the curve was shifted toward less severe degrees of stenosis severity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237797 TI - Effect of restenosis at one previously dilated coronary site on the probability of restenosis at another previously dilated coronary site. AB - The purpose of this study was to determine whether in patients with 2 sites dilated by percutaneous transluminal coronary angioplasty (PTCA), the sites undergo restenosis independently. Although restenosis remains a critical limitation after PTCA, there is little information separating site- and patient dependent determinants of restenosis. In particular, if patients with 2 sites dilated have restenosis at 0 or 2 sites more frequently and at 1 site less frequently than expected by random chance, then patient-related factors may be important in the restenosis process. The source of data was the clinical data base at Emory University. Patients who had previously coronary surgery or PTCA, and those who underwent PTCA in the setting of acute myocardial infarction were excluded. In all, 515 patients with 2 sites dilated undergoing angiographic restudy at 4 months to 1 year after PTCA formed the study population. Site 1 was the first site dilated. At site 1, 224 of 515 sites (45%) were restenotic, and at site 2, 193 (33%) were restenotic. Multiple clinical and angiographic variables were analyzed as possible correlates of restenosis. The most powerful univariate and multivariate correlate of restenosis at either site 1 or 2 was the behavior of the other site. If site 2 was patent, then site 1 was restenotic 28% of the time compared with 69% if site 2 was restenotic. If site 1 was patent, site 2 was restenotic 20% of the time compared with 60% if site 1 was restenotic. This relation was stronger if the 2 sites were in the same coronary artery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237798 TI - Asymptomatic and electrically silent myocardial ischemia during upright leg cycle ergometry and treadmill exercise (clandestine myocardial ischemia). AB - The frequency of "clandestine" (electrically silent and asymptomatic, but scintigraphically evident) myocardial ischemia during treadmill and upright leg cycle ergometric studies was compared in 38 patients with coronary artery disease and reversible ischemia on both thallium-201 scintigraphy and exercise radionuclide angiography. A similar peak double product was attained in both studies. Angina pectoris was significantly more frequent with treadmill exercise than with leg cycling (29 vs 5%, p = 0.0079). An interpretable and positive exercise electrocardiogram occurred in 53% of treadmill tests, but in only 19% of leg cycle tests (p = 0.0025); in a group of 22 patients without scintigraphic ischemia, leg cycle testing had greater specificity (95 vs 64%, p = 0.0248). Ischemia was manifest by symptoms or an abnormal electrocardiogram in 59% of treadmill tests, but in only 18% of leg cycle tests (p = 0.0003). There were no differences between the exercise variables or the degree of scintigraphic abnormalities of the 22 patients with manifest ischemia and the 16 with clandestine ischemia. However, patients with previous coronary artery bypass surgery experienced clandestine ischemia more frequently than did those without bypass surgery (80 vs 33%, p = 0.0103). Thus, clandestine ischemia occurs more frequently during symptom-limited upright leg cycle ergometry (82%) than during treadmill exercise (42%). The symptomatic and electrical manifestations of exercise-induced ischemia have a critical dependence on the type of exercise used. PMID- 8237799 TI - Lack of usefulness of prolonged bleeding times in predicting hemorrhagic events in patients receiving the 7E3 glycoprotein IIb/IIIa platelet antibody. The TAMI Study Group. AB - Hemorrhagic events remain the most worrisome complication in patients receiving drugs that alter hemostasis for treatment of acute coronary syndromes. The 7E3 Fab monoclonal antibody provides a dose-dependent inhibition of platelet aggregation via the glycoprotein IIb/IIIa receptor. This study examines the correlation of hemostatic parameters with bleeding events in patients receiving intravenous 7E3 while enrolled in acute myocardial infarction and high-risk percutaneous transluminal coronary angioplasty pilot studies. Patients with acute myocardial infarction received 100 mg of tissue-type plasminogen activator over 3 hours followed by an escalating intravenous bolus dose of murine 7E3 (0.1 mg/kg [n = 5], 0.2 mg/kg [n = 22], 0.15 mg/kg [n = 13], 0.25 mg/kg [n = 20]). Patients in the high-risk angioplasty trial received a chimeric 7E3 bolus (up to 0.25 mg/kg) with (n = 32) or without (n = 15) intravenous continuous infusion of 7E3 (10 micrograms/min for 6 to 24 hours) after elective angioplasty. Patients in both studies received aspirin therapy (325 mg/day) and partial thromboplastin time-guided (1.5 to 2 times normal) heparin infusion. Bleeding events occurred in 34 of 124 patients (27%). The median template bleeding times (minutes) for patients in the groups with bleeding versus no bleeding events in the trials was 13.5 versus 14 and 30 versus 30, respectively (p = NS). In patients with myocardial infarction, a substantial decline in platelet count at 24 hours was associated with bleeding (p = 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237800 TI - Right ventricular dilatation and remodeling the first year after an initial transmural wall left ventricular myocardial infarction. AB - Left ventricular (LV) remodeling after LV myocardial infarction was described previously. Little is known regarding concomitant adaptation, if any, in right ventricular (RV) volumes after LV infarction. To examine this issue, cine computed tomography was used to determine serial changes in absolute global LV and RV volumes in 27 patients without clinical heart failure during the first year after an initial Q-wave myocardial infarction (14 anterior and 13 inferior). The patient group with anterior wall LV infarction showed progressive increases in LV and RV volumes from hospital discharge to 1 year (end-diastolic volumes +25 and +13%, respectively; and end-systolic volumes +35 and +15%, respectively). In patients with inferior wall LV infarction, both LV end-diastolic and end-systolic volumes increased significantly during the study period (+13 and +15%, respectively). Despite a trend for RV end-diastolic volume to be increased at 1 year, neither end-diastolic nor end-systolic volume increased significantly after hospital discharge following inferior wall LV infarction. Absolute RV end diastolic volume was not significantly different between the infarct groups at any time after infarction. In conclusion, global changes occur in both LV and RV volumes during the first year after an initial infarction regardless of infarct location. The magnitude of these changes was greater after anterior than inferior wall LV infarction. PMID- 8237801 TI - Comparative effects of atenolol versus celiprolol on serum lipids and blood pressure in hyperlipidemic and hypertensive subjects. AB - Antihypertensive drugs may affect serum lipoprotein levels in mixed populations but data in hyperlipidemic patients are scanty. Atenolol versus celiprolol effects on serum lipoproteins were compared in 159 hyperlipoproteinemic hypertensive patients. This was a randomized, double-blind, parallel-group, positive-controlled multicenter trial with centralized lipoprotein laboratory and diet constancy monitoring. Blood pressure reduction and serum lipoprotein and apoprotein levels were monitored for 3 months. Both drugs reduced systolic and diastolic blood pressures. Atenolol had greater effects than celiprolol on diastolic pressure, but effects on systolic blood pressure were not different. Patients receiving atenolol had lower serum high-density lipoprotein cholesterol levels and higher low-density lipoprotein/high-density lipoprotein cholesterol ratios, whereas patients treated with celiprolol showed no contrasting changes. These differences in lipoprotein levels between drug treatment groups were statistically significant at weeks 9 and 12. The difference between drug treatments was also significant if the values of the 9- and 12-week visits were averaged. Patients taking atenolol had statistically significantly higher serum levels of total cholesterol, triglycerides and apoprotein B at 9 weeks. These divergent directional changes were consistent throughout and statistically significantly different between drugs. PMID- 8237802 TI - Improvement in left ventricular function by ablation of atrioventricular nodal conduction in selected patients with lone atrial fibrillation. AB - Left ventricular (LV) function was studied in 30 patients with lone atrial fibrillation (AF) (paroxysmal [n = 27] and persistent [n = 3]) before and after ablation of atrioventricular conduction. In all patients, drug treatment did not control ventricular rate during AF or prevent recurrences of the arrhythmia, or both. LV ejection fraction, and LV end-systolic and end-diastolic, and left atrial dimensions were measured by echocardiography before (mean 7 +/- 10 months, range < 1 to 37) and after (14 +/- 20 months, < 1 to 77) ablation. Before ablation, LV ejection fraction was < or = 50% in 12 patients (group I) and > 50% in 18 (group II). After ablation, LV ejection fraction increased significantly in group I from 43 +/- 8% to 54 +/- 7% (p < 0.0001). There were also significant decreases in LV-end systolic and end-diastolic, and left atrial dimensions. No changes in these parameters were observed in group II. Groups I and II had a significant difference in the duration of AF (group I: mean 11 years, range 8 to 28; and group II: 5 years, 2 to 14) (p < 0.05). No difference was present in age, sex, New York Heart Association functional class for dyspnea, or type of ablation procedure. Thus, some patients with lone AF may show deterioration of LV function, which appears to be related to the duration of the arrhythmia; in these cases, LV function may improve significantly after ventricular rate control is accomplished by ablation of atrioventricular conduction. PMID- 8237803 TI - Long-term effects of theophylline in atrial fibrillation with a slow ventricular response. AB - In 17 patients (aged 78 +/- 9 years) with symptomatic atrial fibrillation and a slow ventricular response not related to drugs, a resting electrocardiogram and 24-hour Holter recording were obtained before and 5 to 6 days after administration of slow-release theophylline (700 mg/day), and successively every 3 months during the long-term phase. Fourteen patients had organic heart disease, and 13 complained of syncope or presyncope, and 4 of asthenia and easy fatigability. At the steady-state evaluation, theophylline significantly increased resting heart rate (HR) by 42%, mean 24-hour HR by 31% and minimal 24 hour HR by 34%. Cardiac pauses > 2,500 ms disappeared or markedly decreased. The daily number of wide QRS complexes increased. Serum theophylline level was 13 +/- 5 ng/ml. During the follow-up period (20 +/- 18 months), the mean daily theophylline dosage was 450 mg and the mean serum theophylline level 9 ng/ml. Seven patients died: 1 because of heart failure, and 6 because of noncardiac death. One patient complained of a syncopal episode during 1 visit. The drug markedly reduced asthenia and easy fatigability. During the long-term phase, HR increased spontaneously in 3 patients, and the treatment was interrupted. In 2 patients, theophylline had to be discontinued because of gastric intolerance. During long-term therapy, HR was similar to that observed at the steady-state evaluation, despite the reduction in daily dosage. The data suggest that theophylline is an effective therapy in most patients with symptomatic atrial fibrillation and a slow ventricular response. PMID- 8237804 TI - A prospective randomized comparison of autodecremental pacing to burst pacing in device therapy for chronic ventricular tachycardia secondary to coronary artery disease. AB - A number of modes of antitachycardia pacing therapies are available in the newer generations of implantable cardioverter/defibrillators. The efficacy of synchronized burst overdrive pacing for the termination of induced and spontaneous monomorphic ventricular tachycardia (VT) was compared with synchronized autodecremental (ramp) pacing in 21 patients who received an implantable antitachycardia pacemaker/cardioverter/defibrillator for treatment of recurrent sustained monomorphic VT. Patients undergoing serial noninvasive VT induction studies after device implantation were prospectively randomized to receive trials of burst or ramp pacing therapies in a crossover study design. Antitachycardia pacing therapies were equally efficacious in treating induced VT (68% for ramp, 76% for burst pacing trials). The efficacy of ramp (93%) and burst (96%) pacing therapies was significantly higher in terminating spontaneously occurring episodes of VT than in terminating induced episodes (p = 0.001). The incidence of tachycardia acceleration was similar for both modes of pacing. The incidence of VT acceleration was lower for spontaneously occurring episodes of VT (0.01%) than for induced episodes of VT (6%, p < 0.01). Thus, antitachycardia pacing is an effective therapy for episodes of monomorphic VT, and the risk of accelerating VT to a hemodynamically unstable form is low. Antitachycardia pacing therapies are more effective against spontaneously occurring episodes than induced episodes of VT. Differences in tachycardia cycle length and duration may contribute to these effects. PMID- 8237805 TI - Follow-up of asystolic episodes in patients with cardioinhibitory, neurally mediated syncope and VVI pacemaker. AB - The occurrence rate of spontaneous asystolic episodes during long-term follow-up in patients with abnormal asystolic responses induced by means of vasovagal maneuvers was evaluated. The heart rate of 23 patients (mean age 64 +/- 12 years; 6 women and 17 men) affected by neurally mediated syncope (mean 4.3 +/- 4.9 episodes) was continuously monitored by a specially designed implanted pacemaker able to detect and store in its memory all asystolic episodes lasting 3 to 6 or > 6 seconds. Asystolic, neurally mediated syncope was diagnosed when a reflex asystole of > 3 seconds was induced during carotid sinus massage (n = 22), eyeball compression test (n = 3) or head-up tilt test (n = 2). During a total of 357 months (mean 15 +/- 7) of monitoring, asystolic episodes occurred in 17 patients (74%): 1,765 episodes of 3- to 6-second (median 3) duration occurred in 14 patients, and 47 episodes of > 6-second (median 2) duration occurred in 11. The actuarial estimates of occurrence of asystolic episodes of > 3 and > 6 seconds were 82 and 53%, respectively, after 2 years of follow-up. Only 12 episodes of 3 to 6 seconds (0.7%), and 20 episodes of > 6 seconds (43%) resulted in presyncopal or syncopal symptoms. Thus, an asystolic response to vasovagal maneuvers predicts the occurrence of spontaneous asystolic episodes during follow up. With few exceptions, spontaneous episodes are asymptomatic and their incidence is low. PMID- 8237806 TI - Effects of long-term oral magnesium chloride replacement in congestive heart failure secondary to coronary artery disease. AB - Magnesium deficiency frequently develops in patients with congestive heart failure and may increase susceptibility to lethal arrhythmias and sudden death via multiple pathophysiologic mechanisms. The effects of peroral magnesium supplementation were investigated in a randomized, double-blind, crossover trial involving 21 patients with stable congestive heart failure secondary to coronary artery disease. All were receiving long-term loop diuretics, and had normal renal function, and low or normal serum magnesium concentrations. Subjects alternately received enteric-coated magnesium chloride (15.8 mmol magnesium per day) and placebo for 6 weeks. Magnesium therapy increased serum magnesium from 0.87 +/- 0.07 to 0.92 +/- 0.05 mmol/liter (p < 0.05), serum potassium from 4.0 +/- 0.3 to 4.3 +/- 0.4 mmol/liter (p < 0.01) and urinary magnesium excretion from 2.82 +/- 0.96 to 4.74 +/- 2.38 mmol/24 hours (p = 0.001). There was no significant change in heart rate or Doppler cardiac index, but mean arterial pressure decreased from 91 +/- 10 to 87 +/- 10 mm Hg (p < 0.05) and systemic vascular resistance from 1,698 +/- 367 to 1,613 +/- 331 dynes s cm-5 (p = 0.047). The frequency of isolated ventricular premature complexes was reduced by 23% (95% confidence interval [CI] 6 to 37%; p < 0.02), couplets by 52% (95% CI 30 to 65%; p < 0.001) and nonsustained ventricular tachycardia episodes by 24% (95% CI 15 to 49%; p < 0.01). Plasma epinephrine decreased from 447 +/- 535 to 184 +/- 106 pg/ml (p = 0.02), but there was no corresponding change in plasma norepinephrine or heart rate variability.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237807 TI - Effects of histamine type 2-receptor antagonists cimetidine and famotidine on left ventricular systolic function in chronic congestive heart failure. AB - Twelve patients with stable congestive heart failure and left ventricular dysfunction were enrolled in a double-blind, randomized crossover trial of famotidine, cimetidine and placebo to determine whether histamine type 2 (H2) antagonists adversely affect left ventricular systolic performance. Two dimensional echocardiograms were obtained at baseline, after 4 days of oral treatment with standard doses of famotidine and cimetidine, and placebo, and at the conclusion of the trial. The baseline mean ejection fraction was 19 +/- 7%. The changes in ejection fraction were +2 +/- 11% (95% confidence interval [CI] -5 to 9%) with famotidine, +3 +/- 10% (95% CI -3 to 10%) with cimetidine, and -3 +/- 7% (95% CI -8 to 2%) with placebo. There were no significant differences in changes in ejection fraction among the 3 experimental treatments (p = 0.22; analysis of variance). The changes in end-systolic wall stress/volume ratios from baseline were +6 +/- 21% (95% CI -6 to 18%) for famotidine, +8 +/- 29% (95% CI -8 to 25%) for cimetidine, and +2% +/- 29% (95% CI -15 to 18%) for placebo (p = 0.69; analysis of variance). No patient had a worsening of symptoms during therapy. Despite previous reports that H2 antagonists may depress left ventricular systolic function, at standard doses these agents result in no decrease in left ventricular systolic function in patients with chronic congestive heart failure. PMID- 8237808 TI - Left ventricular volumes during exercise in normal subjects and patients with dilated cardiomyopathy assessed by first-pass radionuclide angiography. AB - During isotonic exercise, left ventricular (LV) suction and the Frank-Starling law of the heart may have important roles in the enhancement of early LV diastolic filling and in the increase of myocardial contractility, respectively. It remains controversial whether these mechanisms operate in normal subjects or patients with dilated cardiomyopathy. Ten healthy subjects and 10 patients with idiopathic dilated cardiomyopathy who underwent maximal upright bicycle exercise testing were studied. First-pass radionuclide angiography was performed at both rest and peak exercise using a multicrystal gamma camera. In normal subjects, LV end-systolic volume at peak exercise was smaller than during baseline (17 +/- 7 vs 30 +/- 15 ml/m2; p < 0.05), whereas rapid filling volume was greater (52 +/- 16 vs 38 +/- 8 ml/m2; p < 0.01). In patients with dilated cardiomyopathy, both end-systolic (108 +/- 34 to 123 +/- 53 ml/m2; p = NS) and rapid filling (24 +/- 6 to 28 +/- 9 ml/m2; p = NS) volumes did not change from rest to peak exercise. A significant correlation was found between the changes in end-systolic volume at peak exercise and in peak rapid filling rate in normal subjects (r = 0.6; p < 0.05), but not in patients with dilated cardiomyopathy (r = 0.3; p = NS). In normal subjects, end-diastolic volume at peak exercise was similar to that during baseline (78 +/- 14 and 85 +/- 15 ml/m2, respectively; p = NS), whereas in patients with dilated cardiomyopathy, it was greater (164 +/- 50 vs 146 +/- 33 ml/m2; p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237809 TI - Systolic flow velocity profile in the left ventricular outflow tract in persons free of heart disease. AB - It is a common but unsubstantiated belief that blood flows through the left ventricular outflow tract (LVOT) with a spatially uniform velocity. Doppler echocardiography was performed to study in an apically obtained view of the LVOT the profile of subaortic velocities in 79 subjects free of clinical heart disease. The systolic velocities and time intervals were determined in the medial and lateral margins and the center of the LVOT. Peak systolic velocity measured 98 +/- 16 cm/s (mean +/- SD) in the medial margin of the LVOT close to the subaortic ventricular septum, 89 +/- 14 cm/s in the midline of the LVOT, and 79 +/- 14 cm/s in the lateral margin of the LVOT close to the base of the mitral valve (p < 0.001). The respective means of the velocity-time integrals were 19 +/ 3, 17 +/- 3 and 14 +/- 3 cm (p < 0.001). Systolic time intervals determined from the flow signals differed also statistically significantly across the recording sites in the LVOT. The velocity measurements obtained in the center of the LVOT were nearly identical with the values averaged over all measurements across the LVOT. It is concluded that the systolic flow velocity profile in the LVOT is not flat, as has been assumed, but skewed toward faster flow along the subaortic ventricular septum. Measurements made in the center of the LVOT can be used as surrogate for the spatial average velocities. PMID- 8237810 TI - Effects of chronic right ventricular pressure overload on left ventricular diastolic function. AB - Right ventricular (RV) function influences left ventricular (LV) diastolic filling in various clinical and experimental models. The influence of RV systolic function on LV diastolic performance was examined in patients with severe RV pressure overload. Eighty-two patients with pulmonary vascular or parenchymal disease who were referred for heart-lung or lung transplant evaluation were studied. All patients had radionuclide angiography from which RV ejection fraction and LV peak filling rate were measured. Most patients (n = 51) had right sided cardiac catheterization. In 24 patients (group 1), RV ejection fraction was < 30%, whereas in 58 (group 2), it was > 30%. Mean pulmonary artery pressure was greater in group 1 than in 2 (57 +/- 16 vs 34 +/- 20 mm Hg; p < 0.0001). Pulmonary artery wedge pressure was also greater in group 1 than in 2 (14 +/- 9 vs 7 +/- 2 mm Hg; p < 0.0001), whereas peak filling rate was decreased (2.16 +/- 0.88 vs 2.97 +/- 0.79 end-diastolic volumes/s; p < 0.0001). LV ejection fraction was normal in all patients. There was an inverse relation between RV ejection fraction and pulmonary artery wedge pressure (r = 0.45; p < 0.001; SEE 5.3). There was a direct relation between RV ejection fraction and LV peak filling rate (r = 0.49; p < 0.0001; SEE 1.34). In patients with RV pressure overload, RV systolic function is related to LV diastolic performance. This effect is most likely mediated by ventricular interdependence. PMID- 8237811 TI - Impact of heart rate and PR interval on Doppler indexes of left ventricular diastolic filling in an elderly cohort (the Framingham Heart Study). AB - The relations of heart rate and PR interval to Doppler-derived diastolic indexes were examined in 260 men (mean age 75 years) and 462 women (mean age 76 years) from the Framingham Heart Study. Subjects receiving any antihypertensive or cardiac medications were excluded from eligibility; those with mitral stenosis or prosthesis, pacemaker, atrial fibrillation, arrhythmia, left bundle branch block, congestive heart failure, previous myocardial infarction, and technically inadequate Doppler study were also excluded. Peak velocity of early (E) and late (A) diastolic left ventricular (LV) filling, ratio of peak velocities E/A, ratio of time velocity integrals E/A, and atrial filling fraction were studied by multivariable analyses adjusting for age, sex, blood pressure, heart rate and PR interval. Heart rate was a major determinant of all 5 Doppler indexes of diastolic filling; heart rate was inversely associated with peak velocity E, E/A, and time velocity integral E/A, and was directly associated with peak velocity A and atrial filling fraction. PR interval was inversely associated with time velocity integral E/A (p < 0.01) and directly associated with atrial filling fraction. The results were largely unaltered after further adjustment for LV wall thickness, LV end-diastolic diameter and left atrial diameter (in addition to age, sex and blood pressure). Heart rate and PR interval are independent contributors to Doppler-assessed LV diastolic filling in the elderly. The atrial contribution to LV filling depends on its timing in the cardiac cycle and on heart rate. Failure to account for heart rate and PR interval may lead to inappropriate assessment of Doppler diastolic filling. PMID- 8237812 TI - Effects of propranolol and pindolol on cardiac output during extended periods of low-intensity physical activity. AB - The effects of a nonselective beta-adrenergic blocking agent with (pindolol) and without (propranolol) intrinsic sympathomimetic activity properties, compared with placebo-controlled conditions, on metabolic and cardiorespiratory function during long-duration (2 hours) physical activity were examined. After initial cardiorespiratory testing, subjects performed 2-hour walks at 25 and 45% of maximal oxygen consumption (VO2max) under each of the following 3 treatments: pindolol, propranolol and placebo. Medication distribution was randomized and double-blinded. A supine resting blood pressure and electrocardiogram were obtained before each exercise trial. Oxygen consumption, heart rate, stroke volume, cardiac output and blood pressure were determined after 5 minutes of quiet sitting and every 30 minutes during each 2-hour exercise trial. Cardiac output was not significantly different at rest or during exercise, comparing pindolol and propranolol with placebo conditions. Cardiac output tended to decrease over time earlier during propranolol treatment for the 25% VO2max trials in trained normotensive subjects than for the other treatments. Cardiac output decreased at approximately the same time across treatments during the 45% VO2max trials in trained normotensive and untrained hypertensive groups. Finally, owing to the observation that a reduction in cardiac output was delayed or prevented in trained normotensive subjects when compared with that in untrained hypertensives while exercising at 25% VO2max, developing a subject's cardiovascular fitness level may be important in the maintenance of cardiac output during extended periods of low-to-moderate physical activities while under the influence of beta adrenergic blockade. PMID- 8237813 TI - Frequency of oral estrogen replacement therapy in women with normal and abnormal exercise electrocardiograms and normal coronary arteries by angiogram. PMID- 8237814 TI - Prognosis of patients with an isolated fixed thallium-201 defect and no prior myocardial infarction. PMID- 8237815 TI - Prognostic implications of normal exercise SPECT thallium images in patients with strongly positive exercise electrocardiograms. PMID- 8237816 TI - Effects of amlodipine versus diltiazem on morning peak in myocardial ischemic activity in angina pectoris. PMID- 8237817 TI - Plasma lipoproteins are not related to restenosis after successful coronary angioplasty. PMID- 8237818 TI - Stroke complicating percutaneous coronary revascularization. PMID- 8237819 TI - Long-term outcome of patients with asymptomatic restenosis after percutaneous transluminal coronary angioplasty. PMID- 8237820 TI - Low-energy ablation of accessory pathways in cases for which the radiofrequency approach had failed. PMID- 8237821 TI - Oral propafenone as treatment for incessant supraventricular and ventricular tachycardia in children. PMID- 8237822 TI - Canadian Implantable Defibrillator Study (CIDS): study design and organization. CIDS Co-Investigators. AB - The Canadian Implantable Defibrillator Study (CIDS) is an on-going randomized multicenter clinical trial that compares implantable cardioverter-defibrillator (ICD) therapy against amiodarone in patients with prior cardiac arrest or hemodynamically unstable ventricular tachycardia. Eligible patients are equally randomized to receive or not receive an ICD as initial management. Those not receiving an ICD receive amiodarone. All patients are seen in follow-up every 6 months. The primary outcome event cluster is arrhythmic death or any other death occurring within 30 days of therapy initiation. Secondary outcomes are all-cause mortality and nonfatal occurrences of ventricular tachycardia or fibrillation. The goal of the study is to recruit 400 patients over 4 years. All patients will be followed to the end of the year. This will result in an 80% chance of detecting a reduction in arrhythmic death of 58% by ICD if such a difference in truth exists. Recruitment began in October 1990 and 184 patients have been enrolled to date. PMID- 8237823 TI - Preliminary results of the Cardiac Arrest Study Hamburg (CASH). CASH Investigators. AB - Sodium channel blockers and class III antiarrhythmic compounds, as well as beta blockers, have been used in preventing recurrences of sudden cardiac death. In recent years, implantable cardioverter-defibrillators (ICDs) have been used increasingly, but no data from randomized trials comparing antiarrhythmic drug and ICD therapy have been reported in this setting. In 1987, the Cardiac Arrest Study Hamburg (CASH), a prospective, randomized trial, was initiated to compare metoprolol, amiodarone, propafenone, and ICD implantation in patients surviving sudden cardiac death due to documented ventricular tachycardia and/or ventricular fibrillation. The details of the study design and preliminary results are presented herein. The primary endpoint of the study is total mortality. The data reviewed in March 1992, representing a mean follow-up period of 11 months, indicated no significant differences among patients randomized to metoprolol, amiodarone, and ICDs. However, there was a significantly higher total mortality and cardiac arrest recurrence in patients randomized to propafenone compared with those randomized to the ICD treatment limb. The study continues with the deletion of the propafenone treatment limb. PMID- 8237824 TI - Choice and chance in drug therapy of cardiac arrhythmias: technique versus drug specific responses in evaluation of efficacy. AB - Numerous recent advances in pharmacotherapy for arrhythmia have necessitated a reorientation in terms of choice of specific agents, techniques for predicting drug effects, and the endpoints for judging therapeutic efficacy. For the management of ventricular arrhythmias and preventing mortality, several trends are becoming clear. It is unlikely that sodium channel blockers will continue to play a major role, except in patients with structurally normal hearts. Emphasis is shifting way from class I agents to those that act by prolonging repolarization without effect on conduction. These latter agents have been termed pure class III agents and have been developed because of the clinical experience with sotalol and amiodarone. On the other hand, there is compelling evidence that sympathetic inhibition per se (as exemplified by beta blockers) or as an integral component of more complex molecules (e.g., sotalol, amiodarone) is a critical feature of desirable antifibrillatory agents. Thus, compared with D,L-sotalol or amiodarone, pure class III agents are likely to be much less effective and may need to be used in combination with antiadrenergic compounds. Compared with amiodarone, they are likely to induce a higher incidence of torsades de pointes, especially in the case of concomitant diuretic therapy. Therapy guided by programmed electrical stimulation or Holter monitoring is likely to play a diminishing role in the development of antiarrhythmic drug regimens, and thus an antiarrhythmic agent's effectiveness may need to be evaluated against the background of implantable cardioverter-defibrillators or against amiodarone therapy. There is increasing evidence that "guided" therapy may simply identify responders from nonresponders and objective endpoints of therapy may be influenced more by drug-specific responses than by the techniques used for their selection. The data raise the issue whether in the future, therapy for ventricular tachycardia or fibrillation might be chosen empirically but from a limited range of compounds, such as beta blockers, amiodarone, sotalol, and possibly certain pure class III agents that are presently under development. Although it is reasonably certain that there is a need to shift from delaying conduction as a means for treating arrhythmias to one that entails prolongation of repolarization, it remains to be determined what might be the characteristics of an ideal antifibrillatory compound. The greatest promise is the area of complex molecules with a diversity of electrophysiologic actions, as exemplified by amiodarone and similar compounds that have the property of blunting sympathetic excitation. The complexity of their electrophysiologic and pharmacodynamic properties might provide a more favorable match with the vulnerable substrate for reducing electrical instability, thereby preventing ventricular fibrillation. PMID- 8237825 TI - Comparative mechanisms of action of antiarrhythmic drugs. AB - The most widely used classification of antiarrhythmic drugs, formulated by Singh and Vaughan Williams, divides antiarrhythmic agents into 4 categories: (1) sodium channel blockers; (2) sympatholytic agents; (3) drugs that delay repolarization; and (4) calcium antagonists. Despite some controversy regarding its value, the available evidence indicates that this classification relates well to the most important clinically relevant mechanisms of antiarrhythmic drug action. Amiodarone is unique in that it possesses properties belonging to all 4 of the Singh and Vaughan Williams classes; moreover, all 4 properties contribute to the beneficial actions of the drug. Class 1 effects are responsible for amiodarone's ability to slow ventricular tachycardias, making them hemodynamically better tolerated, and are likely important in amiodarone's premature ventricular complex suppressing properties. Class 2 effects may contribute to atrioventricular (AV) node-suppressing actions and may confer protection against sudden death in the postmyocardial infarction population. Class 3 actions contribute to amiodarone's ability to prevent reentrant atrial and ventricular arrhythmias, and may be responsible for a superior ability to maintain sinus rhythm after cardioversion of atrial fibrillation. Class 4 properties contribute to amiodarone's ability to slow the ventricular response in atrial fibrillation and to prevent AV node reentrant arrhythmias. Calcium channel antagonism may also suppress arrhythmias (such as torsades de pointes) caused by early after-depolarizations and contribute to the apparent infrequency of torsades, despite substantial QT prolongation, among patients treated with amiodarone. Consideration of the link between amiodarone's pharmacologic properties and clinical effects illustrates well the various mechanisms of antiarrhythmic drug action. PMID- 8237826 TI - Controlling cardiac arrhythmias by lengthening repolarization: historical overview. AB - An increasing confluence of experimental and clinical data on the gravity of proarrhythmic effects of class I agents has led to a shift to other electrophysiologic classes of agents for treating cardiac arrhythmias. The fact that beta blockers reduce mortality in a variety of subsets of patients has suggested a wider role for this class of agents. Recent investigations have focused on the potential role of sotalol and amiodarone, 2 agents that not only block sympathetic antagonism, but also prolong cardiac repolarization. They have been test drugs in an increasing number of controlled and uncontrolled trials in patients at risk for arrhythmic deaths. Their properties have formed the basis for the hypothesis that arrhythmias may be effectively controlled independently of changes in conduction; they are prototypes of compounds that may favorably influence electrical instability in the myocardium. Amiodarone and sotalol are, however, complex molecules with attendant side effects; therefore, attention is also focusing on compounds that act simply by selective prolongation of cardiac repolarization. These agents have been termed "pure" class III agents. The properties of sotalol, the prototype of class III agents, are of particular interest because it is a racemic mixture with the levo-isomer having 50 times the beta-blocking potency of the dextro-isomer; both have equipotent class III actions. Studies of the antiarrhythmic properties of beta blockers, D- and D,L sotalol, and amiodarone may provide insights into the nature of class III actions. There is clinical evidence indicating that class III drugs exert a varying spectrum of antifibrillatory and proarrhythmic (characterized by torsades de pointes) actions for a given degree of prolongation of repolarization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237827 TI - Antiarrhythmic versus antifibrillatory actions: inference from experimental studies. AB - Pathophysiology of the coronary circulation is a major contributor to altering the myocardial substrate, rendering the heart susceptible to the onset of arrhythmias associated with sudden cardiac death. Antiarrhythmic drug therapy for the prevention of sudden cardiac death has been provided primarily on the basis of trial and error and in some instances based on ill-suited preclinical evaluations. The findings of the Cardiac Arrhythmia Suppression Trial (CAST) requires a reexamination of the manner in which antiarrhythmic drugs are developed before entering into clinical testing. The major deficiency in this area of experimental investigation has been the lack of animal models that would permit preclinical studies to identify potentially useful or deleterious therapeutic agents. Further, CAST has emphasized the need to distinguish between pharmacologic interventions that suppresses nonlethal disturbances of cardiac rhythm as opposed to those agents capable of preventing lethal ventricular tachycardia or ventricular fibrillation. Preclinical models for the testing of antifibrillatory agents must consider the fact that the superimposition of transient ischemic events on an underlying pathophysiologic substrate makes the heart susceptible to lethal arrhythmias. Proarrhythmic events, not observed in the normal heart, may become manifest only when the myocardial substrate has been altered. We describe a model of sudden cardiac death that may more closely simulate the clinical state in humans who are at risk. The experimental results show a good correlation with clinical data regarding agents known to reduce the incidence of lethal arrhythmias as well as those showing proarrhythmic actions. PMID- 8237828 TI - Role of alterations in refractoriness and conduction in the genesis of reentrant arrhythmias. Implications for antiarrhythmic effects of class III drugs. AB - Despite the fact that a number of different electrophysiologic mechanisms are capable of causing cardiac arrhythmias, reentrant excitation has emerged as the most important mechanism causing life-threatening arrhythmias that arise in the ventricles. Pharmacologic therapy of arrhythmias caused by reentry is aimed at preventing the conditions that either facilitate the initiation of the circulating reentrant excitation wave or the conditions that permit its persistence. This involves alterations in either refractoriness or conduction by the drugs. Both atrial and ventricular tachyarrhythmias may follow premature depolarizations that occur at a critical coupling interval to a previous excitation. One desirable property of antiarrhythmic drugs might be to prevent the initiation of reentrant excitation by the triggering premature impulse. Mechanisms are described to show how drugs that prolong the action potential duration (class III antiarrhythmic drugs) might have this effect. It is, however, emphasized that drug effects that have been documented in electrophysiologic studies on normal myocardium might not occur in an arrhythmogenic region that has pathologic alterations, because of changes in the properties of ion channels of the diseased myocardial cells. Antiarrhythmic drugs might also terminate ongoing reentrant excitation by causing block of conduction in the reentrant pathway, at least for one beat. Class III drugs are expected to stop the perpetuation of reentry by prolonging the action potential duration and the refractory period of myocardial fibers in the reentrant circuit to such an extent that the propagating reentrant impulse no longer finds excitable myocardium but blocks in refractory tissue. Therefore, the effectiveness of this drug class to terminate reentry should depend on at least 2 factors: the size of the excitable gap as the reentrant impulse moves around the circuit, which may be related to the mechanism that causes reentry, and the degree to which the drugs can prolong the action potential duration and refractory period at the rapid rates of tachycardia. Each of these factors is discussed with relation to the proposed mechanism of action of drugs that prolong repolarization. PMID- 8237829 TI - Pharmacokinetics of amiodarone: implications for drug therapy. AB - Amiodarone is a complex molecule with multiple pharmacologic properties and a complex electrophysiologic profile. Its disposition kinetics and relation between plasma drug concentration and efficacy can be analyzed using principles identical to those applicable to other antiarrhythmic drugs. However, the drug's affinity for lipophilic tissues, its extremely slow elimination rate, and the likelihood that some of its effects may not be mediated by the usual antiarrhythmic mechanisms confounds traditional pharmacokinetic analysis. Further data that deal with the fundamental mechanisms of action of the drug, in addition to the nature of the relation between dose and uptake into cellular and subcellular fractions and its pharmacologic effects, will be of value in understanding how the drug exerts salutary actions in cardiac arrhythmias. PMID- 8237830 TI - Critical review of the approaches to the prevention of sudden death. AB - The types of cardiac rhythm in patients with sudden death may vary considerably, depending on the underlying cardiac disease. Although ventricular tachyarrhythmias are likely to be the most common causes, a significant proportion of patients may die suddenly due to asystole, electromechanical dissociation, or cardiac rupture. Therefore, the approaches to preventing sudden death may have to be multifactorial. The classic approach of arrhythmia suppression by empiric antiarrhythmic drugs has not resulted in a decrease in sudden death mortality. Although beta blockers have only a modest effect in suppressing arrhythmias, they have been clearly proven to prevent sudden death. Other promising approaches that require further evaluation include modulation of the autonomic balance between the sympathetic and parasympathetic nervous systems (perhaps by beta blockers, exercise training, or low-dose atropine or scopolamine), relief of ischemia by medical or surgical therapy, magnesium supplementation, and mechanical devices, such as the implantable defibrillator. PMID- 8237831 TI - Implications of the ESVEM trial for use of antiarrhythmic drugs that prolong cardiac repolarization. AB - In both the Cardiac Arrhythmia Suppression Trial (CAST) and the Electrophysiologic Study Versus Electrocardiographic Monitoring (ESVEM) trial, class I antiarrhythmic drugs did not appear to perform well, although in the latter trial the class III agent sotalol performed better than 6 class I agents. The results have led some to conclude that class III drugs are now preferred for treatment of ventricular arrhythmias. However, there are a number of reasons why such a conclusion may not be fully justified on the basis of these 2 trials. The data do suggest a need for a critical appraisal and reorientation of drug therapy of life-threatening ventricular arrhythmias. PMID- 8237832 TI - Empiric use of amiodarone and sotalol. AB - Several studies have demonstrated that class I antiarrhythmic drugs do not reduce, and may increase, sudden cardiac death mortality and total cardiac mortality. Because of this, alternative drug choices for antiarrhythmic therapy are necessary. Amiodarone has been demonstrated to be an important and effective antiarrhythmic agent, as has sotalol. The purpose of this article is to review the various indications and possible benefits of the empiric use of these 2 antiarrhythmic agents. PMID- 8237833 TI - The CASCADE Study: randomized antiarrhythmic drug therapy in survivors of cardiac arrest in Seattle. CASCADE Investigators. AB - The Cardiac Arrest in Seattle: Conventional Versus Amiodarone Drug Evaluation (CASCADE) study evaluated antiarrhythmic drug therapy in patients who had survived an episode of out-of-hospital ventricular fibrillation (VF) and who were thought to be at high risk for recurrence of VF. Therapy with empiric amiodarone was compared to therapy with other antiarrhythmic agents, guided by electrophysiologic testing and/or Holter recording. The study evaluated the endpoints of (1) cardiac death, (2) cardiac arrest from ventricular fibrillation with resuscitation, and (3) complete syncope followed by a shock from an automatic implanted defibrillator that restored consciousness. The study comprised 228 patients, 113 treated with amiodarone and 115 treated with conventional antiarrhythmic drug therapy. Most patients had coronary artery disease with a prior myocardial infarction, and one half of the population had a history of congestive heart failure. The mean left ventricular ejection fraction was 35%. Survival was better in patients treated with amiodarone than in patients treated with other antiarrhythmic agents. Patients treated with amiodarone were less likely to receive a shock from an implanted defibrillator, and syncope followed by a shock from a defibrillator was less common in patients treated with amiodarone. However, overall mortality was high, and side effects of therapy were common. Patients treated with amiodarone, even at the low doses used in this study, were still at risk for thyroid dysfunction (both hyperthyroidism and hypothyroidism) and for pulmonary toxicity. PMID- 8237834 TI - Low-dose amiodarone for atrial fibrillation. AB - Concerns about proarrhythmia risk and inefficacy associated with class I antiarrhythmic drugs have revived interest in low-dose amiodarone (maintenance dose 200-400 mg/day) for suppression of atrial fibrillation. In nonrandomized trials of amiodarone for atrial fibrillation refractory to conventional agents, amiodarone has been successful in maintaining sinus rhythm in 53-79% of patients during a mean follow-up of 15-27 months. Intolerable side effects, including pulmonary toxicity, are in the range of 1-12% per year and resolve following amiodarone withdrawal in the majority of cases. Proarrhythmia risk associated with amiodarone, even in the setting of left ventricular dysfunction, is extremely low. In patients with congestive heart failure, in whom other pharmacologic options are limited by proarrhythmia risk and negative inotropism, preliminary experience with amiodarone is especially promising. Randomized trials are needed, directly comparing amiodarone to conventional antiarrhythmic therapy for atrial fibrillation suppression and comparing amiodarone to warfarin for thromboembolism prevention in patients with atrial fibrillation refractory to conventional antiarrhythmic drugs. PMID- 8237835 TI - Secondary prevention after myocardial infarction with class III antiarrhythmic drugs. AB - First-year mortality after myocardial infarction (MI) is high, amounting to 15%. It has been well documented that ventricular arrhythmias late after MI constitute a risk factor for sudden cardiac death. Consequently, several authors undertook attempts to decrease post-MI mortality with antiarrhythmic drugs. Unfortunately, the class I drugs most widely used in clinical practice proved to be ineffective or they even increased the risk of death, as occurred in the Cardiac Arrhythmia Suppression Trial (CAST). So far, only beta blockers, although not particularly effective in controlling ventricular ectopic beats, have been found to decrease first-year mortality after MI by 26-36%. Class III drugs appear to be promising in this clinical setting. Early study with sotalol showed a positive, although statistically nonsignificant, trend toward decreasing mortality. In a more recent trial with amiodarone (Basel Antiarrhythmic Study of Infarct Survival [BASIS]) done in Switzerland, total mortality was reduced (p < 0.05). It should be stressed that the drug was administered at a low dosage level (200 mg/day) to 98 patients and did not cause serious side effects. Similarly encouraging results have been provided by the Polish Amiodarone Study. Amiodarone given to 305 patients at a low dose (200-400 mg/day) reduced first-year cardiac mortality by 42% (p < 0.05). No serious side effects were noticed. Several ongoing trials should further substantiate the impact of this regimen on mortality after MI. PMID- 8237836 TI - Canadian Amiodarone Myocardial Infarction Arrhythmia Trial (CAMIAT): rationale and protocol. CAMIAT Investigators. AB - The Canadian Amiodarone Myocardial Infarction Trial (CAMIAT) is a multicenter, triple-blind, randomized, placebo-controlled trial. Eligible patients are those found on 24-hour ambulatory electrocardiographic monitoring within 6-45 days of acute myocardial infarction to have ventricular premature depolarizations (VPDs) that are frequent (> or = 10/hr) or repetitive (> or = 1 three-beat run of ventricular tachycardia). Consenting patients are randomized to amiodarone or placebo with an oral loading dose of 10 mg/kg/day for 2 weeks; maintenance dose is 300-400 mg/day for 3.5 months, 200-300 mg/day for 4 months, and 200 mg/day for 5 or 7 days/week for 16 months. Patients are followed by alternate telephone and clinical visit at 2-month intervals for 24 months. The principal outcome is a composite of presumed arrhythmic death or resuscitated ventricular fibrillation. Outcomes are determined by an external validation committee. The anticipated rate of arrhythmic death is 7.5% over 2 years; the sample size is 1,200 patients. CAMIAT began in June 1990 and is anticipated to conclude enrollment by June 1994 and follow-up by June 1995. Recruitment rate is about 92% of projected. PMID- 8237837 TI - The European Myocardial Infarct Amiodarone Trial (EMIAT). EMIAT Investigators. AB - The objective of the European Myocardial Infarct Amiodarone Trial (EMIAT) is to assess the efficacy of amiodarone on mortality of patients with depressed left ventricular (LV) function following myocardial infarction (MI). The rationale for the trial is as follows: patients with poor LV function after acute MI have a high sudden cardiac death (SCD) mortality; amiodarone is a successful prophylactic therapy against SCD in patients with ventricular arrhythmias; a number of small studies (Canadian Amiodarone Myocardial Infarction Arrhythmia Trial [CAMIAT] pilot study, Basel Antiarrhythmic Study & Infarct Survival [BASIS], and the Polish Amiodarone Trial [PAT]) of prophylactic amiodarone post AMI have shown a beneficial response attributable to amiodarone. Patients are enrolled between 5 and 21 days after acute MI if LV ejection fraction (assessed by multiple-gated image acquisition nuclear angiography) is < or = 40%. The study group is stratified according to ejection fraction (stratum 1, 31-40%; stratum 2, < 31%). Amiodarone or placebo treatment (blind, randomized) is initiated prior to the discharge of the patient from the hospital and each patient is followed up for the duration of the study, at least 1 year. Recruitment began on November 30, 1990, and will continue for 4 years; > 700 patients are enrolled from > 60 centers (13 countries). The total study mortality (10% at 500 days) and the differential mortality of both strata are as anticipated. Side effects have been infrequent and very few patients have been withdrawn from the study. Trial conclusion is forecast for October 1995. PMID- 8237838 TI - Veterans Affairs congestive heart failure antiarrhythmic trial. CHF STAT Investigators. AB - This is a prospective, double-blind, placebo-controlled trial to determine the effect of antiarrhythmic drug therapy on mortality in patients with congestive heart failure and ventricular arrhythmia. Patients will be assigned to receive either amiodarone or placebo. Eligible patients include those with ischemic and nonischemic congestive heart failure and with > or = 10 ventricular premature beats per hour. All patients must have shortness of breath with minimal exertion or paroxysmal nocturnal dyspnea, a left ventricular internal dimension (LVIDd) by echocardiogram of > or = 55 mm or a cardiothoracic ratio of > 0.5 and an ejection fraction of < or = 40%. All patients will receive vasodilator therapy, unless they find it intolerable. Patients will be entered into the study for 2.5 years and followed for an additional 2 years. Drug therapy will be continued for all patients throughout the entire study unless adverse reactions occur that necessitate individualized treatment. The expectation is that 674 patients will be entered into the study from 25 participating centers. This sample size will allow for the detection of a 33% decrease in 2-year mortality (20% vs 30%) in the treated patients compared with those in the placebo group, with a power of 0.90 and a 2-sided alpha level of 0.05. Intermittent Holter monitoring, radionuclide ventriculograms, pulmonary function tests, echocardiograms, and blood tests, including arterial blood gases, will be required for each patient.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237839 TI - Body weight as essential data in the management of patients with human immunodeficiency virus infection and the acquired immunodeficiency syndrome. PMID- 8237840 TI - Nutrition knowledge and practices of physicians in a family-practice residency program: the effect of an education program provided by a physician nutrition specialist. AB - The purpose of this study was to determine the effects of a nutrition-education program provided by a physician nutrition specialist in a family-practice residency program on residents' and faculty members' nutrition knowledge and nutrition-related patient care, their patients' perceptions of the importance of nutrition, and the physicians' dietary patterns. The nutrition specialist provided recommendations for nutrition patient-care practices to the physicians for 6 mo. Pre- and post-intervention assessments included 1) nutrition exams for physicians and patients, 2) patient questionnaires concerning attitudes towards nutrition, 3) chart reviews, and 4) diet records for physicians. The educational intervention caused a small but statistically significant (P < 0.01) increase in physicians' nutrition-knowledge scores and a significantly higher (P < 0.05) frequency of physicians discussing nutrition and recommending diets for their patients. Effects were greater among residents than among faculty members. The results suggest that a physician nutrition specialist can provide effective nutrition education within a residency program. PMID- 8237841 TI - Effects of a nonenergy fat substitute on children's energy and macronutrient intake. AB - To determine whether children adjusted their energy intake in response to covert manipulations in the proportion of energy from dietary fat, 24-h food intake of 29 2- to 5-y-old children was measured over four 2-d blocks. In this within subject crossover design, in the first three meals of the first day of each block, children consumed foods containing dietary fat or a nonenergy fat substitute, which provided 10% of total daily energy intake. Children compensated for the missing energy; cumulative energy intake differed by only 100 kJ over 2 d. Substitution for dietary fat reduced the percent of energy from fat from 38.7% to 36.4%. Children's intake at individual meals was highly variable (mean CV 24.7%) relative to the variability of total daily energy intake (CV = 8.6%). To produce this pattern, children adjusted energy intake across successive meals. Use of a fat substitute at 10% of energy from dietary fat did not significantly reduce 24-h energy intake. PMID- 8237842 TI - Interactions between growth and nutrient status in school-age children of urban Bangladesh. AB - The relationship between biochemical, anthropometric, and sociodemographic indexes was investigated in 242 children aged 5-12 y from five schools in Dhaka City, Bangladesh. As height-for-age increased so too did the mean serum concentrations of hemoglobin, protein, vitamin A, and zinc; serum copper concentrations were highest in the shortest group. Serum copper concentrations were highest in those with the lowest serum vitamin A concentrations. By multiple regression analysis, family income, age, weight-for-age, hemoglobin, and serum copper were strongly related to serum vitamin A. For every unit change in serum vitamin A there was a 4.92 unit change in hemoglobin, when all the other factors were taken into account. This study shows that there is a complex interaction between concentrations of biochemical indexes of nutritional status and other anthropometric, biochemical, and sociodemographic variables. PMID- 8237843 TI - Decreased rate of stunting among anemic Indonesian preschool children through iron supplementation. AB - Effects of iron supplementation on growth and hematological status of Indonesian anemic preschool children with low weight-for-age were investigated. A treatment group (n = 39) received daily supplements of 30 mg Fe and 20 mg vitamin C, whereas a control group (n = 37) received 20 mg vitamin C only for a period of 2 mo. Supplement allocation was double blind. At the start and finish of the study, body weight, height, food intake, and hemoglobin and serum ferritin concentrations were determined. Only the treatment group showed a significant increase in all hematological values (P < 0.001). Height and weight of all children increased (P < 0.01). Increases in height and height-for-age Z score in the treatment group were larger (P = 0.001) than the increase in the control group. The positive effect of iron supplementation on linear growth was not caused by increased food intake, but seems to be influenced by decreased morbidity. Iron supplementation may be a relatively inexpensive way to help decrease the high prevalence of stunting. PMID- 8237844 TI - Iron status and intake of older infants fed formula vs cow milk with cereal. AB - One hundred four infants were randomly assigned to receive whole cow milk plus iron-fortified cereal (WCM + C) in accord with the previous recommendations of the Committee of Nutrition/American Academy of Pediatrics (CON/AAP); one of two iron-fortified, follow-up formulas; or an iron-fortified infant formula. Mean iron intakes and vitamin C exceeded the recommended dietary allowance in all groups. By 12 mo of age, mean ferritin and mean corpuscular volume were lower in the WCM + C group and significantly more infants had serum ferritin concentrations < 12 micrograms/L. We conclude that infants 6-12 mo of age fed whole cow milk and iron-containing table food are at risk of developing depleted iron stores but not anemia. The iron insufficiency in these infants is not due to inadequate intake of iron or vitamin C, but probably to relatively poor bioavailability of iron in infant cereal. PMID- 8237845 TI - Effect of taurine on synthesis of neutral and acidic sterols and fat absorption in preterm and full-term infants. AB - The effect of dietary taurine on the synthesis of neutral and acidic sterols and fat absorption was investigated in 30 newborn children 2 wk after delivery. The infants were divided into five different groups (n = 6 each) according to their gestational age (GA) and weight for GA, and randomly assigned to receive normal formula or formula supplemented with taurine (479 mumol/L). Neutral sterols, acidic sterols, and fatty acids were determined in formulas and feces by gas liquid chromatography. Only in preterm infants appropriate for GA and small for GA with a mean GA < 33 wk, did taurine supplementation result in lower cholesterol synthesis (-26 +/- 5% and -9 +/- 2%, respectively; P < 0.05) and higher bile acid excretion (100 +/- 35% and 150 +/- 68%, respectively; P < 0.05) and fatty acid absorption (20 +/- 8% and 8 +/- 3%, respectively; P < 0.05). On the basis of these results taurine supplementation is recommended in preterm as well as in small-for-GA neonates < 33 wks of GA who are not on human milk. PMID- 8237846 TI - Hemostatic factors and blood lipids in young Buddhist vegetarians and omnivores. AB - A comparison was made between the hemostatic and lipid profiles of 55 young Chinese Buddhist vegetarians (23 men, 32 women) and 59 Chinese medical students (20 men, 39 women) aged 20-30 y. The modern Buddhist vegetarian diet is high in carbohydrate (63% of energy in men, 58% in women) and has a high polyunsaturated saturated fatty acid ratio, with moderate fat content (25% for men, 30% for women). Rice and soybean proteins are the major protein sources. This study demonstrated that the major beneficial effects of a modern Buddhist vegetarian diet are on blood concentrations of cholesterol, the ratio of apolipoprotein A-I to apolipoprotein B, glucose, and uric acid, but not on most hemostatic factors studied, which included fibrinogen, factor VIIc, factor VIIIc, and plasminogen. Vegetarian men had significantly higher concentrations of antithrombin III than nonvegetarian men. PMID- 8237847 TI - Incorporation of dietary n-3 fatty acids into molecular species of phosphatidyl choline and cholesteryl ester in normal human plasma. AB - To understand the differences in the antiatherogenic actions of eicosapentaenoic acid (EPA, 20:5) and docosahexaenoic acid (DHA, 22:6), we determined their incorporation into molecular species of phosphatidylcholine (PC) and cholesteryl ester (CE) after feeding 12 g marine lipid concentrate/d to six normolipidemic males for 28 d. The time course of incorporation of EPA into plasma PC and CE showed a precursor-product relationship. In contrast, the DHA concentration of CE was markedly lower than that in PC, and the EPA-DHA ratio was 2-6-fold higher in CE than in PC at all time intervals. Three PC species--16:0-20:5, 16:0-22:6, and 18:0-20:5--increased, whereas 18:1-18:2, 18:0-18:2, and 16:0-20:3 decreased. In vitro formation of CE species in plasma by lecithin-cholesterol acyltransferase (LCAT) showed an increased formation of 20:5 CE but not 22:6 CE, indicating that DHA is a poor substrate for LCAT. These results demonstrate a differential incorporation of EPA and DHA into plasma lipids, which may be related to the differences in their biological effects. PMID- 8237848 TI - Can diarrhea induced by lactulose be reduced by prolonged ingestion of lactulose? AB - Twelve healthy volunteers were studied for two test periods, at the beginning of which they ingested a diarrheogenic load (60 g) of lactulose in 350 mL water with 10 g polyethylene glycol 4000 (PEG); the two periods were separated by a lactulose feeding period of 8 d, during which a nondiarrheogenic load (20 g) of lactulose was taken twice daily. The transit time and flow rates of water and lactulose in the distal ileum of four subjects were not different before and after the lactulose feeding period. In the other eight subjects, stool weight and frequency, fecal pH, and fecal outputs of carbohydrates and osmotic moieties after the ingestion of 60 g lactulose dropped significantly (P < 0.05) after the lactulose feeding period, whereas the orofecal transit time and fecal concentrations of beta-galactosidase and lactic acid increased (P < 0.05). We conclude that changes in colonic function induced by prolonged exposure to a nondiarrheogenic amount of lactulose mitigate the severity of the diarrhea because of the larger dose of lactulose. PMID- 8237849 TI - Estimated vitamin intakes of toddlers: predicted prevalence of inadequacy in village populations in Egypt, Kenya, and Mexico. AB - Vitamin intakes of 255 toddlers (aged 18-30 mo) were estimated from food consumption recorded during 1 y at sites in Egypt, Kenya, and Mexico. Mean intakes were compared with requirements standards by using a probability approach to estimate the prevalence of inadequate intakes. There were predicted inadequacies for vitamin A (32%) and riboflavin (20%) in Egypt, vitamins A (68%) and C (63%) and riboflavin (52%) in Mexico, and vitamin B-12 (44%) in Kenya. Vitamin E was inadequate in all diets, but in relation to polyunsaturated fatty acids only the intake in Mexico was low. No diet provided the recommended amount of vitamin D, but its dietary requirement is uncertain. Correlations among nutrient intakes suggest factors that may contribute to reported associations of consumption of animal products with improved growth or development among these children: provision of vitamin B-12 and available minerals, displacement of fiber and phytate-rich energy sources, and increased energy density. PMID- 8237850 TI - Vitamin A supplementation and dietary vitamin A in relation to the risk of xerophthalmia. AB - We examined the effect of 60-mg (200,000-IU) supplements of vitamin A administered every 6 mo on the incidence of xerophthalmia among preschool children who were free of eye symptoms and signs of vitamin A deficiency. We also prospectively studied the relationship of dietary vitamin A intake with the same endpoint. After 18 mo of follow-up, 400 children developed xerophthalmia during 80,104 child-periods of follow-up. Vitamin A supplementation only modestly reduced the risk of xerophthalmia (relative risk 0.88, 95% confidence interval 0.72-1.07, P = 0.19). On the other hand, total dietary vitamin A intake was strongly associated with reduced risk of xerophthalmia; the multivariate relative risk when children in extreme quintiles were compared was 0.38 (95% confidence interval 0.19-0.74; P for trend over quintiles = 0.002). These results emphasize the need for further data on factors that modify the bioavailability of large dose vitamin A supplements. Increased consumption of inexpensive vegetables and fruits is highly likely to reduce significantly the risks of vitamin A deficiency, including nutritional blindness in developing countries. PMID- 8237851 TI - Magnesium bioavailability from human milk, cow milk, and infant formula in suckling rat pups. AB - Little is known about the absorption of magnesium from infant diets. Magnesium bioavailability was evaluated from infant diets that varied by protein and carbohydrate source; magnesium, calcium, and phosphorus contents; and the form of magnesium fortification. Diets were separated into soluble, insoluble, and fat fractions to determine magnesium distribution. Most of the magnesium (> 62%) was found in the soluble fraction. Gel filtration of the soluble fraction from all diets studied showed that > 95% of magnesium is free or associated with low molecular-weight compounds. Distribution of 28Mg and native magnesium in fractions of the diets was similar, thus validating the use of an extrinsic label. In vitro digestion decreased the percent insoluble magnesium from as high as 35% to 2-8%. Whole-body retention of 28Mg-labeled diets in suckling rat pups 4 h after oral intubation ranged from 51% to 92%. No significant differences were found between human milk, cow milk, and infant formula. In conclusion, magnesium from the infant diets studied has high bioavailability, and moderate differences in their composition do not affect bioavailability significantly. PMID- 8237852 TI - Dietary intakes and urinary excretion of calcium and acids: a cross-sectional study of women in China. AB - The relationship between dietary intakes and urinary calcium was examined in a cross-sectional survey of 764 middle-aged and elderly women with markedly different dietary patterns and lifestyles. Urinary calcium was correlated positively with urinary acids, including titratable acid (r = 0.46, P < 0.0001), ammonia (r = 0.42, P < 0.0001), and sulfate (r = 0.52, P < 0.0001). Urinary excretions of calcium and acids were correlated positively with intakes of animal and nondairy animal protein but were correlated negatively with plant-protein intake, possibly because of the alkaline nature of plant foods. Further analyses showed that urinary calcium and acids were associated positively with acid forming foods and were associated negatively with plant foods. These results indicate that under free-living conditions urinary calcium excretion is likely determined by the acid-base status of the total diet, including among other factors the contribution of sulfur amino acids to urinary acid production. PMID- 8237853 TI - Lipids and lipoproteins of malnourished children during early renutrition: apolipoprotein A-IV as a potential index of recovery. AB - Twenty-six children with marasmus and 27 with kwashiorkor were compared with 23 control children of matching ages. Kwashiorkor was characterized by increased phospholipids (NS), low (P < 0.01) apolipoprotein (apo) B-rich LDL, and near normal apo A-I and HDL-C. In children with marasmus apo B (P < 0.02) LDL-C (NS), apo A-I (P < 0.01), and HDL-C (P < 0.001) decreased. Fifteen children in each group were followed for 2 wk. Control values were progressively reached after 2 wk. In the younger children final apo B was higher than in control subjects (P < 0.03) but apo A-I was identical. Apo A-IV, assayed because it correlates with the functional state of intestine, was near normal in children with kwashiorkor and decreased with treatment. In children with marasmus apo A-IV decreased by 50%, increased with treatment in older children, but further diminished in younger children. After 2 wk apo A-IV was significantly lower in all patients than in control subjects. Apo A-IV, by remaining depressed after other variables normalized, seems a good index of nutritional status. PMID- 8237854 TI - Glycosylated hemoglobin concentrations and vitamin E, vitamin C, and beta carotene intake in diabetic and nondiabetic older adults. AB - Studies indicate that large doses of all-rac-alpha-tocopherol in people with diabetes or ascorbic acid in nondiabetic subjects reduces protein glycosylation. The mechanisms by which these nutrients influence glycosylation are poorly understood but may be related to their ability to function as antioxidants. We examined the relationship between glycosylated hemoglobin (GHb) and intake of vitamins E and C and beta-carotene in a population-based sample of middle-aged and older adults participating in the Beaver Dam Eye Study. In people with diabetes, no significant associations were observed between GHb and intake of vitamins E and C and beta-carotene. In people without diabetes, energy-adjusted vitamin C intake was negatively associated with GHb after age and sex were controlled for (dietary, P = 0.02; total, P = 0.04). No significant relationships between GHb and intake of vitamin E and beta-carotene were observed. PMID- 8237855 TI - Prospective analysis of patterns of weight change in stage IV human immunodeficiency virus infection. AB - Weight loss is a major manifestation of infection with the human immunodeficiency virus (HIV). Prospective analysis of weight change was performed in 30 male subjects with stage IV HIV infection over a period of 9-49 mo and weight change events (> 4 kg) related to contemporaneous clinical events. Two distinct patterns of weight loss were observed: episodes of acute severe weight loss and episodes of chronic unremitting progressive weight loss. Thirty-three acute episodes (median 9.1 kg in 1.7 mo) and 23 chronic episodes (13.2 kg in 9.5 mo) were identified. Twenty-seven of 33 (82%) acute weight-loss episodes were associated with nongastrointestinal opportunistic infections and 15 of 23 (65%) chronic episodes with gastrointestinal disease (P < 0.01). Weight loss was neither inevitable nor unremitting. Periods of weight stability (> 4 mo) occurred in 13 individuals (43%); 35 episodes of weight gain were identified, mostly related to recovery from opportunistic infection. These findings have important implications for our understanding of the natural history of weight loss in HIV infection. PMID- 8237856 TI - Herman Award Lecture, 1993: a personal perspective on alcohol, nutrition, and the liver. AB - Alcohol causes primary malnutrition by displacing nutrients in the diet and secondary malnutrition via malabsorption and cellular injury through direct cytotoxicity. Hepatotoxicity results from metabolic disturbances associated with the oxidation of ethanol via liver alcohol dehydrogenase (ADH) and the redox changes produced by the generated NADH (the reduced form of nicotinamide adenine dinucleotide), which in turn affects the metabolism of lipids, carbohydrates, proteins, and purines. Ethanol is also oxidized in liver microsomes by an ethanol inducible cytochrome P450, which contributes to the alcoholic's tolerance and his increased vulnerability to the toxicity of industrial solvents, anesthetics, commonly prescribed drugs, over-the-counter analgesics, chemical carcinogens, and retinoids. Increased acetaldehyde generation, with formation of protein adducts, results in antibody production, enzyme inactivation, decreased DNA repair, impaired utilization of oxygen, glutathione depletion, free radical-mediated toxicity, lipid peroxidation, and increased collagen synthesis. Therapy may eventually improve with the use of supernutrients such as S-adenosyl-L methionine, which replenishes glutathione, restores methylation, and attenuates liver injury, as well as dilinoleoylphosphatidylcholine, which prevents cirrhosis. PMID- 8237857 TI - National Dairy Council Award for Excellence in Medical/Dental Nutrition Education Lecture, 1993: some early beginnings and what now? AB - In 1942 the Department of Nutrition was established at Harvard University jointly in the schools of public health and medicine. It continues to stimulate and expand the teaching of nutrition in these schools, as well as in the school of dental medicine and similar schools in other universities. Nutrition is so broadly involved in health and disease that it should be woven, in an organized fashion, into many of the standard courses in these and other health professional schools. A separate, more detailed course in nutrition may be made available for those who wish to learn more about nutrition. Exposing nutrition quackery and other types of nutrition misinformation should be part of any program in nutrition education, particularly in schools of medicine, public health, and dentistry. Stimulating the teaching of nutrition in schools for health professionals requires the enthusiastic support of the dean and faculty as well as adequate financial support. PMID- 8237858 TI - Fatty acid composition of colostrum of Turkish women. PMID- 8237859 TI - Comparison of intake and blood concentrations of micronutrients. PMID- 8237860 TI - Nutrition and human physiological adaptations to space flight. AB - Space flight provides a model for the study of healthy individuals undergoing unique stresses. This review focuses on how physiological adaptations to weightlessness may affect nutrient and food requirements in space. These adaptations include reductions in body water and plasma volume, which affect the renal and cardiovascular systems and thereby fluid and electrolyte requirements. Changes in muscle mass and function may affect requirements for energy, protein and amino acids. Changes in bone mass lead to increased urinary calcium concentrations, which may increase the risk of forming renal stones. Space motion sickness may influence putative changes in gastro-intestinal-hepatic function; neurosensory alterations may affect smell and taste. Some or all of these effects may be ameliorated through the use of specially designed dietary countermeasures. PMID- 8237861 TI - Use of dual-energy x-ray absorptiometry in body-composition studies: not yet a "gold standard". PMID- 8237862 TI - Resting metabolic rate and diet-induced thermogenesis: a methodological reappraisal. AB - The variability in resting metabolic rate (RMR), diet-induced thermogenesis (DIT), and fuel utilization rates as well as the impact of several factors on RMR and DIT were assessed in several studies with a total of 103 males and females. The intraindividual CV of RMR and respiratory quotients was 5-6%. The intraindividual variability in DIT and fuel utilization rates was substantially higher. RMR did not change from morning to afternoon. The menstrual cycle phase did not affect RMR and DIT. DIT after mixed meals of 1.3-2.6 MJ could be assessed with good accuracy in 3 h. It is concluded that the low reproducibility of DIT implies that sample sizes of < 10 individuals with one measurement per subject and per treatment have power levels < 80% of assessing true, relatively large (50%) treatment effects or between-group differences in DIT. PMID- 8237863 TI - A critical evaluation of heart rate monitoring to assess energy expenditure in individuals. AB - The prediction of energy expenditure (EE) from minute-by-minute heart rate (HR) recording was evaluated in 40 female workers, with emphasis on the inter- and intraindividual variation of this method. The results show that the relationship between EE and HR varied greatly between and within subjects. The inter- and intraindividual CVs were 14-18% and 11-20%, respectively. The poor limits of agreement in EE (mean difference +/- 2 SD) between group and individual calibration curves indicate that estimated EE from group curves is inferior to estimated EE from individual curves. Therefore, it is concluded that to have the best estimates of individual EE, only individual calibration curves should be used. At different occasions these curves should be generated again. Because the limits of agreement in EE between individual curves based on 18 and 9 different activities were wide (-2399 to 1817 kJ/16 h), it is preferable to have a wide range of different activities in the individual calibration procedure. PMID- 8237864 TI - Effects of infused sodium acetate, sodium lactate, and sodium beta hydroxybutyrate on energy expenditure and substrate oxidation rates in lean humans. AB - Infusion of sodium acetate in lean humans results in a decrease in respiratory exchange ratio, which may be advantageous in patients with respiratory failure. However, this potential decrease in respiratory work was observed to be offset by significant thermogenesis. The metabolic effects of sodium acetate, sodium lactate, and sodium beta-hydroxybutyrate, infused at a rate of 20 mumol.kg-1.min 1 for 3 h, was monitored in six healthy human volunteers. Respiratory exchange ratio decreased from 0.85 +/- 0.02 at baseline to 0.75 +/- 0.02, 0.75 +/- 0.02, and 0.80 +/- 0.02, after acetate, lactate, or beta-hydroxybutyrate, respectively (P < 0.05 for each). Acetate produced a larger thermic effect (22.7% of energy infused) than did lactate (16.3%) or beta-hydroxybutyrate (13.6%). Thus, sodium salts of organic acids may potentially decrease the respiratory requirements by decreasing the respiratory exchange ratio. However, this effect is partially offset by the thermic effect of these substrates. The maximal doses and safety of these anions during larger infusion periods remain to be determined. PMID- 8237865 TI - Adaptive role of energy expenditure in modulating body fat and protein deposition during catch-up growth after early undernutrition. AB - After 10 or 30 d of growth arrest due to undernutrition during the early weaning period, rehabilitated rats fed ad libitum showed the phenomenon of catch-up growth, ie, gains in body weight and body protein were greater by 50% and 25%, respectively, (P < 0.001) than those of controls with similar starting body weight [ie, weight-matched (WM) controls]. These increases, however, were entirely dependent on the higher food intake because they failed to occur when food intakes were maintained at similar amounts to those for WM controls. In contrast, independent of whether the rehabilitated groups were spontaneously hyperphagic relative to WM controls or made normophagic by pair-feeding to WM controls, the gross energetic efficiency was increased (P < 0.01) and body fat gain was more elevated (2-2.5 fold, P < 0.001) during the first 2 wk of refeeding compared with WM controls--differences that were uninfluenced by the duration of growth arrest. Taken together, these studies suggest that the often reported impressive gains in body fat during recovery from malnutrition may result not only from unbalanced diets or excess dietary intake, but also from a transitory enhancement in the efficiency of food utilization and a shift in energy partitioning in favor of an acceleration for the replenishment of fat stores. PMID- 8237866 TI - Evaluation of a gastric delivery system for iron supplementation in pregnancy. AB - The present investigation was undertaken to assess the efficacy of oral iron supplementation during pregnancy by using a gastric delivery system (GDS). Three hundred seventy-six pregnant women between 16 and 35 y of age and 14 and 22 wk gestation were selected if mild anemia was present (hemoglobin concentration 80 110 g/L). The participants were randomly assigned to one of three study groups given no iron, two FeSO4 tablets (100 mg Fe) daily, or one GDS capsule (50 mg Fe) daily. Blood was obtained initially and after 6 and 12 wk for measurement of red blood cell and iron indexes, including serum transferrin receptor. There was a significant and comparable improvement in hematologic and iron-status measurements in the two groups of women given iron whereas iron deficiency evolved in women given no iron supplement. We conclude that by eliminating gastrointestinal side effects and reducing the administration frequency of an iron supplement to once daily, a GDS offers significant advantages for iron supplementation of pregnant women. PMID- 8237867 TI - Effects of energy and protein intakes on pregnancy outcome: an overview of the research evidence from controlled clinical trials. AB - This overview summarizes the evidence from controlled clinical trials bearing on the effects of energy and protein intakes during gestation on the outcome of pregnancy, based on meta-analyses of reports contained in the Oxford Database of Perinatal Trials. Trials of nutritional advice to increase energy and protein intakes, and of balanced energy and protein supplementation, have demonstrated only modest increases in maternal weight gain and fetal growth, even in undernourished women, and no long-term benefits to the child in terms of growth or neurocognitive development. Neither balanced isoenergetic protein supplementation nor high-protein supplementation appears beneficial to either mother or infant and may even impair fetal growth. The same can be said for energy and protein restriction in pregnant women who are overweight or exhibit high weight gain. Future trials should concentrate on prevention of preterm birth and fetal and neonatal death; such trials will require substantially larger sample sizes than those used in the past. PMID- 8237868 TI - Effects of maternal nutritional status and maternal energy supplementation on length of postpartum amenorrhea among Guatemalan women. AB - To investigate the extent to which better maternal nutrition leads to reduction in length of postpartum amenorrhea, multivariate-logistic and linear-regression analyses were applied to data on 339 mother-infant pairs from the longitudinal Guatemalan Four Village Study, 1969-1977. Maternal triceps skinfold thickness was negatively associated with length of amenorrhea when infant supplementation (a proxy for reduced suckling) was accounted for. However, its effect was small: amenorrhea was only 0.5 mo shorter among women at the 75th percentile than among those at the 25th, equivalent to less than even one additional child during the women's reproductive years. Maternal supplementation was not associated with length of amenorrhea when infant supplementation was controlled. This is in contrast to previous studies in which breast-feeding or infant supplementation was not controlled. These results suggest that infant, not maternal, supplementation influences length of postpartum amenorrhea, and that maternal nutritional status has minimal influence. PMID- 8237869 TI - Selenium status of infants is influenced by supplementation of formula or maternal diets. AB - Plasma selenium of infants fed proprietary formula was significantly less than that in infants fed human milk. Addition of selenite to the formula (0.253 mumol Se/L) increased plasma selenium and activities of glutathione peroxidase (GPx) and total peroxidase (Px). However, erythrocyte selenium decreased significantly during the 12-wk study in infants receiving human milk or formula with or without supplemental selenite. Infants fed human milk from women receiving 0 or 200 micrograms supplemental selenium as selenomethionine or selenium-enriched yeast had plasma selenium that paralleled changes in their selenium intake. Plasma GPx and Px activities were unrelated to human milk selenium intake. Milk from women given either selenium supplement prevented the decline in infant erythrocyte selenium. Results of these studies suggest that the method of feeding modifies the infant's apparent selenium status and that the molecular form of selenium provided and/or its interaction with other milk constituents are determinants of infant selenium status. PMID- 8237870 TI - Selenium status of lactating women is affected by the form of selenium consumed. AB - The impact of providing selenomethionine (2.7 mumol Se) or selenium-enriched yeast (2.9 mumol Se) on the selenium status of lactating and nonlactating women with customary intakes of approximately 1.3 mumol Se/d was studied. Plasma selenium declined in unsupplemented lactating women but not in nonlactating women. Selenomethionine increased plasma selenium in both lactating and nonlactating women whereas selenium-enriched yeast increased plasma selenium only in nonlactating women. Erythrocyte selenium concentration was not significantly modified by lactation. Plasma glutathione peroxidase (GPx) activity decreased with duration of lactation in unsupplemented women and selenomethionine or selenium-enriched yeast supplementation prevented the decline. Milk selenium declined markedly for 20 wk after parturition in unsupplemented women. Selenomethionine significantly increased milk selenium concentrations whereas selenium-enriched yeast prevented a decline. These results clearly show that the source of selenium provided to lactating women can significantly influence selected indexes of selenium status, including milk selenium concentration. PMID- 8237871 TI - Dependence of human milk essential fatty acids on adipose stores during lactation. AB - The relationships between essential fatty acid (EFA) composition of colostrum, mature milk, and white adipose tissue (WAT) were examined on days 5 and 30 postpartum in 24 healthy French mothers. Fatty acid composition was assessed by capillary gas chromatography. In WAT, the proportion of individual polyunsaturated fatty acids (PUFAs) did not change during lactation and was greater (18:2n-6) or lower (18:3n-3, long-chain PUFAs) than values found in colostrum or mature milk (P < 0.04). The 18:2n-6 content and the ratio of 18:3n-3 to 18:2n-6 correlated between WAT and colostrum (r = 0.52 and r = 0.57, respectively) or mature milk (r = 0.64 and r = 0.65, respectively). These relationships agree with an expected qualitative effect of WAT fatty acid composition on interindividual variability of milk parent EFA content. The decrease in the long-chain PUFA content observed from colostrum to mature milk and the concomitant occurrence of a precursor-product relationship between the linoleate and its long-chain PUFA are consistent with the mobilization of a preformed long-chain PUFA pool during early lactation. PMID- 8237872 TI - Renal adaptation to dietary carnitine in humans. AB - Carnitine homeostasis in humans is maintained by dietary carnitine intake, a modest rate of endogenous carnitine synthesis, and efficient conservation of carnitine by the kidney. To assess the effect of dietary carnitine on the efficiency of carnitine reabsorption in humans, rates of carnitine excretion and reabsorption, indexed to the glomerular filtration rate, were determined over a range of plasma free and total carnitine concentrations in 12 strict vegetarians before and after dietary carnitine supplementation (0.248 mmol/d). This amount of dietary carnitine supplementation did not significantly increase plasma carnitine concentration and did not alter the glomerular filtration rate. At normal physiological plasma carnitine concentrations, the rate of carnitine excretion was increased and the rate of carnitine reabsorption was decreased by carnitine supplementation. We conclude that the kidney adapts to carnitine intake by reducing the efficiency of carnitine reabsorption. PMID- 8237873 TI - Comparative efficacy of n-3 and n-6 polyunsaturated fatty acids in modulating ventricular fibrillation threshold in marmoset monkeys. AB - Programmed electrical stimulation in anesthetized marmoset monkeys was used to examine relative antiarrhythmic efficacies of dietary n-3 and n-6 polyunsaturated fatty acids (PUFAs) from fish and plant oils. Diets contained 31% of energy (en%) as fat, comprising 15 en% saturated fat and 7 en% PUFAs, obtained by blending sheep fat with sunflower seed (SF/SSO) or fish oil (SF/FO) and a base diet. After 16-wk feeding, ventricular fibrillation (VF) was inducible in 6 of 10 animals on each diet under control conditions. The VF threshold (VFT) was significantly elevated in the SF/FO group (33.3 +/- 3.1 mA; n = 6) compared with the SF/SSO group (14.3 +/- 4.9 mA; n = 6). VFT, reduced during acute myocardial ischemia with 10 of 10 animals inducible per diet, remained significantly higher with SF/FO feeding. The SF/FO diet contained 3.8 en% as n-3 PUFAs, which was incorporated as 31% of myocardial membrane fatty acids. Dietary n-3 PUFA reduced vulnerability of normal or ischemic myocardium to arrhythmias in a nonhuman primate. PMID- 8237874 TI - Requirements for indispensable amino acids in adult humans: longer-term amino acid kinetic study with support for the adequacy of the Massachusetts Institute of Technology amino acid requirement pattern. AB - Twenty young men received an L-amino acid diet, supplying 140 mg N.kg-1 x d-1 and patterned as in the Egg diet for 1 wk, and then for 3 wk either a pattern based on international recommendations (modified FAO diet; n = 7), our new amino acid requirement pattern (MIT diet; n = 7), or the egg pattern (Egg diet; n = 6). At the end of the initial week, at 1 and 3 wk with the three experimental diets, and after 3 d after return to the Egg diet, an 8-h continuous intravenous infusion with [1-13C]leucine (3 h fast, 5 h fed while subjects received hourly meals supplying the equivalent of one-twelfth the daily intake) was conducted. After 3 wk with the different diets, mean daily leucine balances were lower (P < 0.01) with the FAO diet (-160 mumol.kg-1 x d-1) than with the MIT diet (-15 mumol.kg-1 x d-1). Together with changes in plasma amino acid profiles [eg, methionine increased (P < 0.05) during feeding with the FAO and Egg diets but not with the MIT diet; increased proline concentrations during the fed state (P < 0.05) with the FAO diet but not with the MIT or Egg diets] we interpret these findings to indicate that the FAO diet is not capable of maintaining amino acid homeostasis, as is the case with the MIT and Egg diets. PMID- 8237875 TI - Low serum 25-hydroxyvitamin D concentrations and secondary hyperparathyroidism in middle-aged white strict vegetarians. AB - The vitamin D status of vegetarians was studied in the winter. The groups studied were strict vegetarians (G1), lactovegetarians (G2), lactoovovegetarians eating some fish (G3), and vegetarians who were taking vitamin D supplements or who had been exposed to abundant sunlight during the last 6 mo (G4). A group of healthy women served as control subjects (C). The serum 25-hydroxyvitamin D [25(OH)D] concentration was significantly lower, the serum intact parathyroid hormone (S iPTH) concentration significantly higher, and the dietary vitamin D intake significantly lower in G1 than in C. S-iPTH correlated negatively with serum 25(OH)D and dietary calcium intake. In conclusion, white strict vegetarians are at risk of vitamin D deficiency, at least in the winter, primarily because of a low dietary vitamin D intake, despite a normal sunlight exposure in summer. Low serum 25(OH)D concentrations are accompanied by high S-iPTH concentrations, which also are affected by a low calcium intake. The effect of these changes on bone health remains to be evaluated. PMID- 8237876 TI - Zinc absorption in healthy elderly humans and the effect of diet. AB - Absorption of a zinc stable isotope was measured on two consecutive occasions in nine young and eight elderly healthy men aged 24-40 and 70-83 y, respectively. A zinc stable-isotope label (0.8 mg 70Zn) was added to a test meal of either high or low zinc bioavailability, depending mainly on phytic acid content. Zinc absorption from the high-bioavailability test meal was not significantly different (P > 0.05) in the young (38.9 +/- 9.8%, mean +/- SD) and elderly (35.0 +/- 10.9%) subjects. Zinc absorption from the low-bioavailability test meal was 40% and 43% lower, at 23.4 +/- 10.2% and 19.8 +/- 6.1% in these young and elderly men, respectively. Again, no significant effect of age was found. These results show that aging does not lead to nutritionally relevant changes in zinc absorption and in the effect of dietary inhibitors on zinc absorption. Thus, zinc absorption ability seems to be preserved in healthy elderly people, at least until the age of 80 y. PMID- 8237877 TI - Meal-induced changes in plasma, erythrocyte, and urinary zinc concentrations in adult women. AB - We attempted to determine whether there is a limit to the transient, meal-induced decline in plasma zinc and whether there is a concomitant increase in erythrocyte and erythrocyte membrane concentrations. Premenopausal women participated in a 17 h fasting trial, a one-meal trial with breakfast at 0700, and a three-meal trial with meals at 0700, 0900, and 1100. During fasting, plasma zinc increased 9%; it decreased 11% and 19% in the one- and three-meal trials, respectively (P < 0.001). A limit to the decline in plasma zinc was reached after the second meal in the three-meal trial. Erythrocyte, erythrocyte membrane zinc, and serum calcium and phosphorus concentrations did not change significantly during the three trials. Serum glucose concentrations were weakly related to plasma zinc concentrations, suggesting that the postprandial decline in plasma zinc is associated with the metabolic changes caused by food intake. PMID- 8237878 TI - Effects of dietary zinc restriction on postprandial changes in plasma zinc. AB - The principal objective of this study was to determine the effect of a short period of severe dietary zinc restriction on both prebreakfast and postbreakfast plasma zinc concentrations. After an overnight fast, plasma zinc was measured in five normal adults before and at 30-min intervals for 6 h after a standard zinc free test meal. This test was performed after a week on a zinc-adequate diet and repeated after a week of severe dietary zinc restriction. Postabsorptive premeal plasma zinc did not change after a week of severe dietary zinc restriction but the posttest meal decline was significantly greater and the maximum decline less variable (15-23%). It is concluded that postprandial plasma zinc is more sensitive than prebreakfast plasma zinc to dietary zinc depletion. Recent zinc intake is one factor that determines the magnitude of the postprandial net efflux of zinc from the plasma compartment. PMID- 8237879 TI - Variations in iron-status measures during the menstrual cycle. AB - To determine whether normal physiologic changes associated with hormone fluctuations over the menstrual cycle affect concentrations of iron-status indicators, we examined data from 1712 women aged 18-44 y from the Second National Health and Nutrition Examination Survey (NHANES II) after adjusting for potential confounders. Adjusted mean values of hemoglobin (Hb), transferrin saturation (TS), and serum ferritin (SF) were lowest for women whose blood was drawn during menses and highest for women examined in luteal or late luteal phase of the menstrual cycle (Hb = 130 vs 133 g/L; TS = 21.2% vs 24.8%, P < 0.01 for both; and SF = 17.2 vs 24.0 micrograms/L, P < 0.05). The prevalence estimate of impaired iron status was significantly higher for women whose blood was drawn during the menstrual phase than for women whose blood was drawn during the luteal and late luteal phases. Our findings suggest that the phases of the menstrual cycle affect the concentration or values of iron-status indicators. These cyclic variations in indicators of iron status are a potential source of error when iron status is assessed in large population surveys that include women of reproductive age. PMID- 8237880 TI - Resting metabolic rate in sickle cell disease. PMID- 8237881 TI - An increase in atherogeneity on a very-low-energy diet? PMID- 8237882 TI - Late luteal dysphoria and food intake. PMID- 8237883 TI - A modified method of hydrodensitometry in young children. PMID- 8237884 TI - Conversion of insulin units to SI units. PMID- 8237885 TI - Carrageenan: an asset or detriment in infant formula? PMID- 8237886 TI - Tumor necrosis factor and resting energy expenditure during the acquired immunodeficiency syndrome. PMID- 8237887 TI - Effects of sham and real auricular needling: implications for trials of acupuncture for cocaine addiction. AB - We conducted a single-blind study (n = 48) comparing subjective evaluations of needles inserted into sham and real auricular acupuncture points. Both ears were needled concurrently, one in sham sites and the other in active points used in the treatment of cocaine addiction. Subjects then completed a questionnaire rating the intensity of five sensations in each ear, and also attempted to identify which ear received sham and which ear received real acupuncture. Results showed that real points were more painful than sham; there were no other overall differences. Subjects' ability to identify which ear received sham and which ear received real acupuncture did not rise above the level of chance. PMID- 8237888 TI - Beneficial effect of acupuncture on adult patients with asthma bronchiale. AB - In a retrospective study 17 patients with long-standing history of asthma bronchiale were treated with acupuncture at the outpatient unit of the Department of Anaesthesia and Intensive Care in the University Hospital of Vienna. The subjective effectiveness of the treatment was determined using a standard questionnaire, which was sent to the patients' homes half a year after starting acupuncture treatment. Over 70% of our patients reported a significant improvement of their ailments after ten weeks of treatment as well as half a year after starting acupuncture. PMID- 8237889 TI - A comparison of the traditional Chinese versus transpositional zangfu organ association acupoint locations in the horse. AB - The traditional Chinese (TC) and transpositional (TP) methods of animal acupoint location result in different acupoint charts. Representative TC and TP equine zangfu organ shu-association acupoint charts are compared to each other and to a human chart. Despite their differences, practitioners of both methods appear to achieve equally effective therapeutic results--a phenomenon termed "traditional Chinese/transpositional equal efficacy" (TTEE). Common veterinary acupuncture practices, traditional Chinese medical theory, spinal cord anatomy, and a preliminary equine "association segment" chart are proposed to explain TTEE. The differences between the charts indicate that all documented animal acupoint locations should be explicitly described. PMID- 8237890 TI - Moxibustion for correction of breech presentation: a clinical study with retrospective control. AB - In this study we treated a group of women during pregnancy by moxibustion on point 67 B (Zhiyin), to obtain inversion of fetuses in breech presentation. Comparison is made with a control group drawn retrospectively from clinical files at a regional hospital. The aim of the study is to identify the ideal population (in terms of parity and gestational age) to be included in a randomized controlled trial. PMID- 8237891 TI - Pharmacological actions of "kyushin," a drug containing toad venom (3): Effects on experimentally induced arrhythmia. AB - The pharmacological effects of the toad venom-containing drug "kyushin" on aconitine- and thyroxine-induced arrhythmia in guinea pigs, on the conduction system in Langendorff preparations of rabbit hearts and on the autonomic nervous system in cats were studied. "kyushin" significantly inhibited the aconitine induced arrhythmia after intraduodenal administration (i.d.) with 80 mg/kg, and the thyroxine-induced arrhythmia with 40 mg/kg i.d. Although "kyushin" itself did not affect the conduction system with 30 mg/ml of the maximal concentration being able to be prepared, bufalin and cinobufagin as constituents of toad venom produced inhibition with 0.3 mg/ml and 1 mg/ml, respectively. The decrease in heart rate induced by electrical stimulation to the parasympathetic nerve (vagus nerve) was potentiated by "kyushin" at 30 mg/kg i.d. The anti-arrhythmic effects of "kyushin" may be attributable to both possible inhibitory effect on the conduction system and potentiating effect on the parasympathetic nervous system. PMID- 8237892 TI - Effects of kuei-pi-tang on cellular immunocompetence of gamma-irradiated mice. AB - Kuei-Pi-Tang is a kind of traditional Chinese medicine which has been suggested to have therapeutic effects on hemato-deficient disease and radiation injuries. In order to further investigate its protection function, this study is focused on the efficacy of kuei-pi-tang on cellular immunocompetence of gamma-ray irradiated mice. ICR strain male mice were chosen and divided into several groups for their different treatments of 4 Gy gamma-ray whole body irradiation and kuei-pi-tang administration. After the treatments, six to eight mice from each group were sacrificed on days 1, 5, 12, 19, 26 and 33. The body and splenic weights of mice by different treatments were measured and the splenic cells separated thereafter. The changes of cellular immunocompetence in mice following treatments were measured by 3H-thymidine incorporation. The results revealed that 4 Gy of gamma ray irradiation inhibits the increases of body and splenic weights and exerts a pronounced inhibitory effect on the incorporative rates of 3H-thymidine in the splenic lymphoid cells which have been stimulated by mitogens. kuei-pi-tang administration seems to increase the recovery of the cellular immunocompetence, especially for the treatment of kuei-pi-tang administered with the concentration of 20 mg/20 g body weight after gamma-ray irradiation. PMID- 8237893 TI - Effects of rehmanniae radix water extract on renal function and renin secretion rate in unanesthetized rabbits. AB - This study was undertaken to explore the effects of different preparations of Rehmanniae Radix on renal function and renin secretion rate. Steamed Rehmanniae showed a decrease in urine volume, renal hemodynamics, and urinary excretion of electrolytes confined to the infused kidney, and was followed by an increase in urinary excretion of electrolytes from both kidneys. Dried Rehmanniae showed a decrease in urine volume, glomerular filtration rate, and urinary excretion of electrolytes, while raw Rehmanniae showed only an increase in urinary excretion of electrolytes. Steamed Rehmanniae and dried Rehmanniae decreased renin secretion rate, but raw Rehmanniae tended to increase renin secretion. These results suggest that the Rehmanniae Radix preparations have different effects on renal function and renin secretion rate. PMID- 8237894 TI - Preliminary study on the anti-radiation effect of jen-sheng-yang-yung-tang. AB - Six to seven week old male mice of ICR strain were exposed to different doses of x-rays to determine if jen-sheng-yang-yung-tang could be a modifier in the elimination of radiation damage. Colony forming units of bone marrow cells in the spleen (CFUs) were measured before and after x-ray irradiation with intraperitoneal injection of 10 mg/20 g or 20 mg/20 g body weight of jen-sheng yang-yung-tang, once a day for seven consecutive days. The recovery of CFUs and hemocytes counts by 4 Gy irradiation with jen-sheng-yang-yung-tang administration was faster for a concentration of 20 mg/20 g than 10 mg/20 g. The measurement of 10-day CFUs showed an increase of radiotolerance in the treatment of 20 mg/20 g administration before x-ray irradiation. The injection of jen-sheng-yang-yung tang accelerated the recovery of hemocyte counts in mice irradiated with 4 Gy x ray; the effect was especially profound for leukocytes with 20 mg/20 g jen-sheng yang-yung-tang administration after irradiation. PMID- 8237895 TI - Two Class II, division 1 patients with congenitally missing lower central incisors. AB - Although orthodontic treatment objectives and procedures for apparent protrusion of the maxillary teeth vary among orthodontists and specific cases, the differences are even greater where there is disharmony of jaw relationship between the maxilla and the mandible. The two cases presented in this article resemble each other in appearance, but the growth patterns and reactions to appliance, as well as treatment progress, are quite different. The initial excessive overjet seemed equally severe. Treatment was started at the same age, both patients were eldest daughters, both had a convex type facial pattern and an abnormal perioral muscle function with mouth breathing. In addition to the marked overjet, deep bite, and Class II molar relationship, lower central incisors were missing, and second molars had not erupted. They both had a steep curve of Spee, disharmony between the upper and lower arch forms, and slight diastemata. The facial appearance has been improved significantly in both, and patient satisfaction is high. A Kloehn-type cervical headgear was used in both cases. Case 1 showed improvement in the relationship between the maxilla and the mandible with reduction of the overjet. However, in Case 2, the sagittal relationship became worse, and an open bite developed. The obvious question is why the big difference in treatment response? What role did clearing of the nasopharyngeal airway play? PMID- 8237896 TI - Effect of enamel bleaching on the bonding strength of orthodontic brackets. AB - Various whitening/bleaching systems have been introduced on the market, including an over-the-counter external bleaching material, containing carbamide peroxide. It has been suggested that the process might alter the enamel surface structure in a manner similar to acid etching. Orthodontists are interested in determining whether the process will alter the adhesive characteristics of orthodontic bonding systems. The results of this study indicate that the use of 10% carbamide peroxide did not result in significant changes in the shear debonding strength of orthodontic brackets. PMID- 8237897 TI - Initial stress induced in periodontal tissue with diverse degrees of bone loss by an orthodontic force: tridimensional analysis by means of the finite element method. AB - This study was undertaken to determine the stress that appears in tooth, periodontal ligament and alveolar bone, when a labiolingual force of 100 gm is applied in a labiolingual direction in a midpoint of the crown of an inferior digitalized canine, and its changes depending on the degree of loss of the supporting bone. The analysis of tensions was carried out by means of the finite element method (FEM) for a normal case and after reducing the periodontal support bone 2, 4, 6, and 8 mm. Three-dimensional images in false color in which intensity of tensions and its areas of extension are generated. Special attention was paid to changes at level D (apical transversal section) to which maximum, mean, minimum, and Von Mises tensions are calculated. After applying the labiolingual force in the canine, a progressive increase of the stress in the labial and lingual zones of the tooth, periodontal membrane and alveolar bone was observed when the alveolar bone was reducing. In the mesial and distal zones, no compensating forces appear, which could provoke a tooth rotation during the tipping movements. PMID- 8237898 TI - A comparison of surgery and orthodontics in "borderline" adults with Class II, division 1 malocclusions. AB - From a pool of 108 former patients, discriminant analysis was used to identify a homogeneous borderline prognostic subgroup of 27 adult orthodontic and 26 adult surgical Class II patients who, before treatment, were similar with respect to the characteristics on which the orthodontic/surgical decision appears to have been based. The fact that some had been treated orthodontically, whereas others had been treated surgically, was taken as empirical evidence that the patients in this stratum were equally susceptible to the two treatments and that the actual choice was largely a function of whose office they happened to contact. The former orthodontic patients were recalled an average of 7.1 years after treatment, the former surgical patients, 4.7 years after surgery. Each subject was evaluated with respect to skeletal and dental stability, profile esthetics, and temporomandibular function. Although there were dramatic differences in the nature of the correction (dental versus skeletal), both groups of patients generally thought that their profiles had been improved by treatment. As judged by data generated from visual analogue scales, the mean difference between the orthodontic and surgical patients' evaluations of their treatments was small and nonsignificant. Moreover, the "borderline" Class II orthodontic and surgical patients showed no significant differences in craniomandibular function and incisor stability. There was, however, one profound difference between treatments: 3 of the 26 surgical patients underwent extensive relapse, probably as a result of condylar resorption. From the standpoint of estimated probabilities and utilities for the various outcomes, the present results imply that orthodontics would be the better choice for the borderline adult Class II patient, whereas surgery would be appropriate for the more severely affected patient. PMID- 8237899 TI - A longitudinal cephalometric study of transverse and vertical craniofacial growth. AB - Longitudinal growth changes were studied from posteroanterior cephalometric radiographs of 25 male subjects from 4 to 25 years of age and 25 female subjects from 4 to 20 years of age who had Class I skeletal and dental patterns. Growth for males continued past age 18 years for all skeletal measurements, except for maxillary width. Growth for females was completed by 17 years for all skeletal measurements. At 6 years of age the transverse measurements had a greater percentage of the adult size completed than vertical measurements for both males and females. Gender differences at age 6 years were in the mean widths for the cranium, face, and maxilla. At age 12 years the differences were in cranial width, maxillary width, and maxillary and mandibular intermolar width (6-6). There were gender differences at age 18 years for all the variables, except for nasal width and mandibular intermolar width (6-6). Regression lines provided strong-to-moderate predictive equations to determine the size of most of the measurements at age 18 years, if the value at age 6 years is known. PMID- 8237900 TI - Increase in vertical dimension alters mechanical properties and isometric ATPase activity in guinea pig masseter. AB - To study the changes in mechanical and metabolic properties associated with an increase in the vertical dimension of the face, isometric tension, isometric ATPase activity, unloaded shortening velocity (Vmax), and the tension transients in response to step stretches in length were measured at constant levels of various Ca2+ activations in glycerinated masseter muscles (75 to 150 microns in diameter and about 3 mm long) from normal and bite opened (6 mm increase in the vertical dimension, period of 1 week) guinea pigs. The isometric tension increased sigmoidally with an increase in Ca2+ concentration in both preparations. However, the bite opening shifted the pCa-relative tension relationship in the direction of increasing Ca2+ required for activation, and the pCa at Km (Ca2+ concentration required to develop half maximum tension) was 6.40 +/- 0.08 in control and 6.27 +/- 0.05 in the bite opened preparations. The maximum isometric tension at the saturating Ca2+ concentration was greater in the bite opened preparation. The ATPase activity changed almost in parallel with the tension (i.e., Ca2+ concentration) in both preparations. However, the tension cost (ATPase activity/tension) was significantly (p < 0.05) lower in the bite opened preparation, compared with the control. The Vmax at the saturating Ca2+ concentration was lower in the bite opened preparations. The tension transients in response to step stretch in length were characterized by four distinct phases; the first phase of an immediate tension increase coincident with the stretch, the second phase of a rapid quasi-exponential tension reduction, the third phase of a minute tension increase and the fourth phase of a slow tension reduction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8237901 TI - Dynamics of tissue changes found after mechanical loading of the rat incisor. II. A three-dimensional longitudinal study of the histopathologic aspects. AB - The study comprised 60 lower left rat incisors subjected to 2 weeks of loading (19.7 +/- 1.6 gm) and 5 control teeth. After the loading period, 10 rats were killed with the springs in situ, and the remainder were killed in groups of 5 at intervals of 1 through 10 weeks after the springs were removed. The teeth were cross-sectioned serially (2 microns). The distance of each section from the apex and the time of its tissue formation in relation to load removal were calculated. Bone remodeling and dental and periodontal trauma were plotted according to their location on the tooth axis and circumference. Vigorous bone remodeling continued throughout the observation period, apposition and resorption sites intermingling around the tooth. Directionality, as expressed by the resorption/apposition ratio, for the formerly tensed and stressed areas was 0.61 +/- 0.07 and 1.79 +/- 0.52, respectively. However, in some of the groups directionality was negligible or reversed. New lesions of dentin (folding and resorption) were formed for 6 weeks after loading, whereas new damage to the enamel organ and enamel matrix appeared until week 9. The integrity of the PDL deteriorated for the first 8 weeks, as evidenced by edema, cell loss, scarring, and collagen fiber disorientation. Necrosis occurred throughout the recovery period. Fiber to bone attachment was disrupted by formation of big sinusoids adjacent to the bone. The frequency of most lesions decreased toward the end of the observation period. It is suggested that tissue recovery proceeds in subsiding cycles of tissue injury/tissue repair, a process that is much more prolonged than is generally believed. PMID- 8237902 TI - A redefinition of bilateral sagittal osteotomy (BSO) advancement relapse. PMID- 8237903 TI - Reflections on an academic career in teaching and research. PMID- 8237904 TI - Prepare, the world is coming to the end! PMID- 8237905 TI - The standard of care and expert testimony. PMID- 8237906 TI - Genetics: the quiet revolution in science and medicine. Implications for research on child health issues, education of health professionals, and the new preventive and curative medicine. PMID- 8237907 TI - The Human Genome Project and the future of medicine. AB - The Human Genome Project is an international research effort the goal of which is to analyze the structure of human DNA and to determine the location of the estimated 100,000 human genes. Another component of the program is to analyze the DNA of a set of nonhuman model organisms to provide comparative information that is essential for understanding how the human genome functions. The project began formally in 1990. In this report, we summarize the rapid progress that has already been made; the impact that the resources already developed by the Human Genome Project have had on the ability of investigators to identify and isolate human genes, particularly those associated with disease; and the promise that the project offers for profoundly altering our approach to medical care, from one of treatment of advanced disease to prevention based on the identification of individual risk. PMID- 8237908 TI - State of the art of biochemical genetics. PMID- 8237909 TI - Human cytogenetics. A current overview. AB - Chromosomal abnormalities are the basis for a substantial proportion of human morbidity and mortality. During the past 35 years, the field of human cytogenetics has helped to elucidate the etiology of many congenital malformation/mental retardation syndromes. Through adaptation of technological advances and integration of molecular biological techniques, cytogenetics continues to contribute significantly to our knowledge of clinical genetics, chromosomal fine structure and function, gene mapping, and prenatal diagnosis. This review outlines the basic concepts, recent findings, and current laboratory approaches to cytogenetic diagnosis. PMID- 8237910 TI - Gene therapy. PMID- 8237911 TI - Modern diagnosis and treatment of Gaucher's disease. PMID- 8237912 TI - The state of the art of dysmorphology. AB - The art of dysmorphology is simply a more detailed historical and clinical evaluation of a potentially dysmorphic individual. It requires precise observation and collating of all physical features, followed by interpretation, categorization, and diagnosis. It also requires an understanding of morphogenesis combined with an appreciation of form. Clinicians who practice dysmorphology well have certain personality traits that aid them in their pursuit of a diagnosis. Dysmorphology is at its apex of clinical maturation, but scientific discoveries and technology have begun to explain how and why malformations arise and why syndromes vary. Nevertheless, expertise in dysmorphology remains a clinical skill attainable by all who desire it. This visual diagnostic modality is inexpensive and often quick. Continued emphasis in training of dysmorphology in genetic fellowship programs will be critical to producing well-balanced clinical geneticists. PMID- 8237913 TI - Genetic diagnosis and treatment. Ethical considerations. PMID- 8237914 TI - Telediagnostic conferencing. PMID- 8237915 TI - Sickle cell anemia. Beta s gene cluster haplotypes as genetic markers for severe disease expression. AB - Identification of the beta s gene cluster haplotype and alpha-gene status provides a useful tool for the detection of high-risk patients with sickle cell anemia. Analysis of the relationship of the long-term clinical course to the above parameters has revealed that those with the haplotype designated Senegal have decreased severity, those with the Benin haplotype have intermediate severity, and those with the Central African Republic (CAR) haplotype have the most severe clinical expression. Further modulation of the clinical course occurs with the coinheritance of alpha-thalassemia-2. In both Africa and the United States, the CAR beta s haplotype increased the risk (relative risk, 2.25; 95% confidence interval, 1.41 to 3.87) of developing a complication and death at an early age. Detection of the CAR haplotype identifies the child with sickle cell anemia at risk for a rapid rate of progression of sickle-induced microvasculopathy, ultimately leading to irreversible organ damage during the first three decades of life. In patients with the CAR haplotype, potential curative therapy, such as bone marrow transplantation or gene insertion, should be seriously considered during childhood, before organ failure is clinically evident. PMID- 8237916 TI - The genetics of infantile hypertrophic pyloric stenosis. A reanalysis. AB - OBJECTIVE: To determine whether the existing family data for infantile hypertrophic pyloric stenosis (IHPS) are sufficient for the purposes of establishing the mode of inheritance of this condition. DESIGN: Reanalysis of the familial aggregation patterns exhibited by IHPS, using data from several published family studies. CONCLUSIONS: Due to several limitations of the available family data for IHPS, the results of this analysis should be interpreted cautiously. Within the context of these limitations, the familial recurrence pattern among monozygotic cotwins and more remote relatives of IHPS probands was found to be inconsistent with generalized single major locus inheritance. The familial recurrence pattern of IHPS is, however, compatible with multifactorial threshold inheritance or the effects of multiple interacting loci. Under a model of multiple interacting loci, no single locus can account for more than a fivefold increase in the risk to first-degree relatives of IHPS probands. In contrast to several earlier reports, this analysis does not support the existence of a maternal factor that contributes to the risk of IHPS in the offspring of affected females. PMID- 8237917 TI - Velo-cardio-facial syndrome. Intrafamilial variability of the phenotype. AB - We describe a mother and son with velo-cardio-facial syndrome (VCFS) in whom cytogenetic and DNA molecular studies demonstrate an interstitial deletion of the long arm of chromosome 22. Although these two individuals manifest the typical facial and cognitive features of VCFS, they are discordant for the cardiovascular and palatal anomalies, which are seminal manifestations of the disorder. Previously, this degree of phenotypic variability had not been well appreciated within a single family segregating the VCFS deletion. A review of other familial cases of VCFS suggests that the family described in this article is not atypical. Because a microdeletion would be expected to be inherited without alteration within individual families, the phenotypic variability observed in these families appears to be an intrinsic quality of the syndrome and not wholly due to genetic heterogeneity. PMID- 8237918 TI - The North American Collaborative Study of Maternal Phenylketonuria. Status report 1993. AB - Neonatal screening for phenylketonuria (PKU) has created an unexpected problem as females with PKU are reaching childbearing age. Surveys have revealed that maternal phenylalanine blood concentrations above 1200 mumol/L are associated with microcephaly, mental retardation, congenital heart defects, and intrauterine growth retardation among their offspring. It is estimated that as many as 3000 hyperphenylalaninemic females may be at risk for producing these fetal abnormalities. To examine this problem, the North American Maternal PKU Collaborative Study has been developed to evaluate the efficacy of a phenylalanine-restricted diet in reducing fetal morbidity. Preliminary findings indicate that phenylalanine restriction should begin before conception for females with PKU planning a pregnancy. Dietary control should maintain maternal blood phenylalanine levels between 120 and 360 mumol/L and should provide adequate energy, protein, vitamin, and mineral intake. Pregnant hyperphenylalaninemic females who achieved metabolic control after conception or by the 10th week of pregnancy had a better offspring outcome than anticipated. PMID- 8237919 TI - A study of the physical, behavioral, and medical phenotype, including anthropometric measures, of females with fragile X syndrome. AB - The physical features of fragile X, including a long face, prominent ears, and hyperextensible joints, are present in affected males and females. Cytogenetically negative heterozygotes have been considered to be unaffected by the fragile X mental retardation-1 (FMR-1) gene. This study investigated the penetrance of the FMR-1 gene in cytogenetically negative but DNA-positive heterozygotes with a premutation (cytosine guanine guanine [CGG] amplification in the 50 to 200 repeat range), compared with carriers with a full mutation (> 200 CGG repeats) and control subjects. One hundred thirty-nine women with normal IQs between the ages of 18 and 45 years were studied. All underwent cytogenetic and DNA testing to determine their fragile X carrier status. A medical history-taking and a physical examination, including selected anthropometric measurements, were performed. Results indicate that the FMR-1 mutation mildly affects the physical phenotype of individuals even in the premutation state, although less dramatically than more affected heterozygotes. Carriers with a premutation differed significantly from control subjects in overall physical index score and in the anthropometric measure of ear prominence. These results suggest a phenotypic impact of the FMR-1 mutation even at the 50 to 200 CGG repeat length. PMID- 8237920 TI - X-linked lymphoproliferative disease. PMID- 8237921 TI - Williams syndrome, Down syndrome, and cognitive neuroscience. PMID- 8237922 TI - Pitfalls in genetic counseling for childhood disorders: the pediatrician's role. PMID- 8237923 TI - Acute lymphoblastic leukemia in a 46,XY/47,XYY mosaic male: clonal origin of leukemia in the XY-bearing stem-cell line. PMID- 8237924 TI - High-grade dysplasia in Barrett's esophagus: an evolving clinical dilemma. PMID- 8237925 TI - Portal hypertensive gastropathy. PMID- 8237926 TI - Holding a hand is often as important as examining one. PMID- 8237927 TI - Review of alcoholic hepatitis, and its treatment. PMID- 8237928 TI - Surgical management of high-grade dysplasia in Barrett's esophagus. AB - The role of surgery in patients with Barrett's esophagus and high-grade dysplasia is controversial. The aims of this study were to determine the prevalence of unsuspected early cancer and to evaluate surgical outcome in a cohort of patients with high-grade dysplasia. Records of all 16 patients who underwent esophagectomy for high-grade dysplasia from 1986 to 1991 were reviewed. All had preoperative endoscopy with no gross evidence of carcinoma, and none had a preoperative diagnosis of intramucosal or invasive carcinoma. Intramucosal carcinoma was found in six (38%) resection specimens. There were no cases of invasive carcinoma or lymph node metastases. One patient (6%) died 3 months postoperatively. The remaining patients are alive without evidence of recurrent cancer (range of follow-up, 2-68 months). Early postoperative complications occurred in seven patients (44%). Late complications occurred in 11 patients (73%). Anastomotic strictures accounted for seven of the 11 (64%) late complications. Complications were successfully managed conservatively in all but two patients. One required laryngectomy for chronic aspiration and another required a gastrojejunostomy for gastric outlet obstruction. Intramucosal carcinoma that had been unsuspected is frequently found in patients with Barrett's esophagus and high-grade dysplasia. Mortality associated with esophagectomy is low, and perioperative complications can usually be managed conservatively. Esophageal resection is indicated in appropriately selected patients with Barrett's esophagus and high-grade dysplasia. PMID- 8237929 TI - Effects of transcatheter splenic arterial embolization on portal hypertensive gastric mucosa. AB - OBJECTIVES: The aim of this study was to address the effects of transcatheter splenic arterial embolization on portal hypertensive gastric mucosa. METHODS: Thirty patients with portal hypertension and hypersplenism were included in this study. Seventeen patients underwent transcatheter splenic arterial embolization and 13 did not. The latter served as controls. Gastric mucosal hemodynamics was assessed by reflectance spectrophotometry which provides gastric mucosal hemoglobin content and oxygen saturation. Portal hypertensive gastropathy was also classified into three major categories (i.e., absent, mild, or severe). These examinations were performed before and 3 months after the procedure (i.e., splenic embolization or control). RESULTS: Control had no significant changes on either gastric mucosal hemoglobin content or oxygen saturation. In contrast, splenic embolization induced a 11% reduction in gastric mucosal hemoglobin content (p < 0.01), although gastric mucosal oxygenation did not increase. Further, there was a significant difference in the rate of improvement of portal hypertensive gastropathy (splenic embolization vs. control; 71% vs. 8%, p < 0.05). CONCLUSION: These results suggest that transcatheter splenic arterial embolization has a beneficial effect on portal hypertensive gastric mucosa in patients with hypersplenism. PMID- 8237930 TI - A scoring system to predict rebleeding after endoscopic therapy of nonvariceal upper gastrointestinal hemorrhage, with a comparison of heat probe and ethanol injection. AB - We prospectively and randomly compared heat probe and ethanol injection in 80 patients with major nonvariceal upper gastrointestinal hemorrhage who were bleeding actively or had endoscopic stigmata associated with a high risk for rebleeding. We also attempted to predict which patients would rebleed within 72 h after successful endoscopic therapy, using a three-component scoring system. Heat probe and ethanol injection proved to be similar in efficacy and safety. Active bleeding was controlled with equal success with heat probe and ethanol injection (92% vs. 82%), and there was no difference in the rebleeding rate (11% vs. 13%). The scoring system was useful in predicting which patients would rebleed. Significant differences were seen in the mean values of all three scores, and specific cut-offs in the pre-endoscopy and post-endoscopy scores predicted patients who rebled. High likelihood ratios and post-test probabilities for rebleeding were found for the number and severity of concurrent illnesses, but not for endoscopic stigmata, implying that the excess risk associated with stigmata is eliminated after effective endoscopic therapy, and clinical factors become the primary determinants of rebleeding. PMID- 8237931 TI - Effects of intravenous nitroglycerin and metoclopramide on intravariceal pressure: a double-blind, randomized study. AB - In search of an effective agent for decreasing intravariceal pressure, 27 patients with portal hypertension and large esophageal varices were studied. Thirteen patients received iv nitroglycerin (150-micrograms bolus), and 14 received iv metoclopramide (20-mg bolus). Continuous measurement of variceal pressure and systemic hemodynamics was carried out. Compared with the baseline, the variceal pressure was significantly reduced 10 min after nitroglycerin injection (20.23 +/- 3.1 vs. 14.6 +/- 4.8 mm Hg, p < 0.05): the percentage reduction was 27 +/- 25.4%. With metoclopramide, variceal pressure decreased by 12.7 +/- 22%; from 21.9 +/- 5.6 to 19.1 +/- 7.7 mmHg (p = NS). There was no significant alteration in the heart rate and mean arterial pressure with either drug. Our results thus suggest that in the dosages used, intravenous nitroglycerin effectively and safely reduces the variceal pressure. Further studies using a combination of nitroglycerin and metoclopramide need to be considered. PMID- 8237932 TI - Physiologic and psychologic characteristics of an elderly population with chronic constipation. AB - OBJECTIVES: To characterize colorectal physiologic parameters and psychological profiles in subjects > or = 60 yr with chronic constipation who have not sought medical advice. METHODS: Colorectal physiologic studies and psychologic profiles were obtained in 18 self-identified constipated subjects and 17 nonconstipated control subjects. Constipation was defined as two or fewer bowel movements/week and/or defecatory difficulty for at least 6 months. All subjects underwent colonic transit with radiopaque markers, anorectal manometry and completed a self rated inventory assessing psychological distress (SCL-90-R). RESULTS: Slow colonic transit occurred in five constipated subjects. Pelvic floor dyssynergia and/or increased rectal compliance were found in nine subjects versus only two controls. One-third of constipated subjects scored in the top 10th percentile for global psychologic distress with higher scores for somatization, depression, and anxiety than control subjects. Neither physiologic nor psychologic abnormalities were predictable on the basis of patterns of bowel complaints. CONCLUSIONS: Colorectal physiologic and psychologic dysfunctions are common in elderly constipated subjects who do not seek medical attention for bowel dysfunction. PMID- 8237933 TI - Clarithromycin as monotherapy for eradication of Helicobacter pylori: a randomized, double-blind trial. AB - Current regimens to eradicate Helicobacter pylori usually consist of metronidazole plus a bismuth compound, as well as a third agent such as tetracycline. Such regimens are not ideal because organisms may be metronidazole resistant, side-effects occur, and compliance is often poor. This randomized, double-blind study was designed to assess the ability of clarithromycin, a new macrolide antimicrobial, as monotherapy to eradicate H. pylori. Thirty-seven healthy volunteers who were H. pylori positive by 13C-urea breath test plus histology and/or culture completed 14 days of oral therapy with clarithromycin in one of three dosages. Eradication, defined as all three tests negative at 4-6 wk after the end of therapy, was achieved in 2/13 (15%) with clarithromycin 500 mg bid, 4/11 (36%) with 1000 mg bid, and 7/13 (54%) with 500 mg qid. Isolates of H. pylori were resistant to clarithromycin prior to therapy in 12% of subjects, and became resistant during therapy in 21% of subjects. Taste perversion, the most common side effect, resulted in one subject terminating therapy. CONCLUSIONS: Whereas clarithromycin is a promising antimicrobial in the eradication of H. pylori, it is not sufficient to be used as monotherapy. PMID- 8237934 TI - Minimum analysis requirements for the detection of Helicobacter pylori infection by the 13C-urea breath test. AB - BACKGROUND: In an attempt to identify the minimum criteria for scoring the 13C urea breath test, we examined test results from 96 healthy subjects, 270 duodenal ulcer patients undergoing treatment to eradicate Helicobacter pylori, and 1000 consecutive breath tests analyzed in our laboratory. METHODS: The 13CO2/12CO2 isotope ratio in a baseline sample was compared with that of samples collected at 20, 30, 40, and 50 min post-dosing. A positive test was defined as an average increase in the ratio of 6/1000 over baseline. Values for a two-sample analysis (baseline and one other) and for a single-sample analysis were extracted from the data and compared with the original analysis outcome. RESULTS: Test results were negative for 186 patients and positive for 84. The two-sample method had high specificity (94-99/1000), sensitivity (95-99/1000), and positive predictive value (88-97/1000). False-positive results occurred most often with samples collected at 20 min and least often with the sample at 40 min (11 and 1, respectively). Analyses based on a single sample collected beyond 20 min, in which samples with an isotope ratio > or = -15/1000 versus the standard were defined as positive, gave comparable specificity (98-99/1000), slightly reduced sensitivity (92-94%), and positive predictive value (96-99%). The analyses of 1000 consecutive breath tests yielded approximately 1% false-positive and false-negative results by the two-sample method; the single-sample method yielded a 2% false-positive and false negative result. CONCLUSION: The 13C-urea breath test can determine H. pylori infective status from a single breath sample collected at least 30 min post-dose in which the absolute 13C abundance is > or = -15/1000 versus the standard. PMID- 8237935 TI - Successful application of extracorporeal liver perfusion: a technology whose time has come. AB - We have used extracorporeal liver perfusion (ECLP) to aid in the management of three patients with fulminant hepatic failure (FHF). Organs were used for ECLP only if they would have gone otherwise unused through United Network for Organ Sharing. In all three patients treated, serial serum bilirubin and arterial ammonia values trended toward the normal range. The neurologic examinations improved dramatically in two patients, and metabolic function of the extracorporeally perfused livers was unequivocally demonstrated by the clearance of theophylline in the last two patients. Two patients ultimately had successful liver transplants, whereas the third patient failed to improve neurologically despite evidence of metabolic function by the extracorporeally perfused liver, and died 7 days after ECLP was discontinued, from pulmonary and renal failure. These studies suggest that, 30 yr after initial clinical trials, ECLP can be applied safely without the need for arterial access 1) as a bridge to transplantation, 2) to assess whether patients in FHF will benefit from improved hepatic function and therefore transplantation, and 3) potentially, to evaluate the "usability" of questionable donor organs. PMID- 8237936 TI - Cytomegalovirus infection of the gastrointestinal tract in non-AIDS patients. AB - Cytomegalovirus infection is usually reported in immunocompromised patients. In this study, 19 patients with cytomegalovirus (CMV) infection of the alimentary tract were reviewed, none of whom had acquired immunodeficiency syndrome. The patients' sex distribution was similar, and their ages ranged from 22 to 84 yr (mean = 61 yr). Only four patients had underlying diseases resulting in immunocompromise. The most common presentation was gastrointestinal bleeding, which occurred in 11 of the 19 patients (58%). Two patients had an unusual presentation of obstructive jaundice due to exuberant growth of granulation tissue at the ampulla of Vater. The majority of the lesions (n = 23) occurred in the large intestine and duodenum. Sixteen of these 23 lesions (69.6%) showed ulceration. Of these 16 lesions, six were diagnosed macroscopically as malignant by the endoscopists. It was observed histologically that a predominant epithelial distribution of the CMV inclusion bodies was associated with no or only mild inflammation, whereas a predominant endothelial distribution of the inclusion bodies was associated with ulcerative, severely inflamed lesions. From these observations, we conclude that ulceration probably is due to an ischemic process resulting from narrowing of capillary lumens by swollen endothelial cells affected by CMV infection. Concomitant infection by Candida species was found in only one patient. One patient had coexistent ulcerative colitis while another also suffered from graft-versus-host disease after bone marrow transplantation. In five of the eight cases with follow-up biopsies, the CMV inclusion bodies disappeared in subsequent biopsies. To conclude, a high degree of alertness is required for the histological diagnosis of unexpected CMV infection of the alimentary tract, in order to warrant correct treatment for the patients. PMID- 8237937 TI - A randomized, controlled trial of recombinant alpha-interferon therapy for chronic hepatitis B. AB - OBJECTIVES: To evaluate the effect of recombinant alpha-interferon in chronic hepatitis B. METHODS: Patients were stratified at entry according to their serum aspartate aminotransferase (AST) values, randomized to receive alpha-interferon (alfa-2b, 10 million units three times weekly) or to be untreated controls for 16 wk. Effect of therapy on levels of hepatitis B viral (HBV) DNA and aminotransferase activities in serum and hepatitis B e antigen (HBeAg) status was monitored. RESULTS: Forty-seven patients entered the trial; 11 of 25 (44%) patients receiving interferon responded by clearing HBeAg and HBV DNA within 6 months, compared to one of 22 (5%) controls (p < 0.05). Among those with serum AST values < 100 U/L, 33% responded and among those with AST values > 100 U/L, 60% responded. Within the 6-month study period, 36% of treated patients had normal serum alanine aminotransferase (ALT) values, and 16% had cleared hepatitis B surface antigen (HBsAg) from serum, whereas none of the controls had normal ALT values or had lost HBsAg. Interferon was stopped early in three patients (6.5%), and dosage was reduced in a further 16 patients (35%) because of adverse effects. Predictive factors for a response were the pretreatment serum ALT and AST activities. CONCLUSIONS: alpha-Interferon therapy (three times weekly) is relatively well tolerated and is effective in clearing HBeAg and HBV DNA in approximately one-third of treated patients. PMID- 8237938 TI - Differences in manometry and 24-H ambulatory pH-metry between patients with and without endoscopic or histological esophagitis in gastroesophageal reflux disease. AB - In this study, our objective was to examine prospectively, by endoscopy and histology of the esophageal mucosa, the severity of reflux esophagitis and any possible correlation between endoscopic and histological findings on the one hand, and manometric and 24-h ambulatory pH-metry measurements on the other. Forty-two patients with gastroesophageal reflux were prospectively examined. The results were compared with those of 18 healthy controls. Methods used were: 1) upper alimentary endoscopy and grading of severity of esophagitis, 2) esophageal mucosa biopsies, to estimate severity of esophagitis on histology, 3) standard esophageal manometry, by using a water perfused catheter with four side holes, and 4) standard 24-h ambulatory esophageal pH-metry. RESULTS: The severity of esophagitis as determined by both endoscopy and histology was significantly inversely related to the amplitude (p < 0.001) and duration (p < 0.01) of esophageal peristalsis at 5 cm proximal to LES; it was significantly related to the pH-metry total composite score (p < 0.001 for endoscopy, p < 0.05-0.01 for histology), the total reflux time (p < 0.001 for endoscopy, p < 0.01 for histology), the duration of longest reflux episode (p < 0.001 for endoscopy, p < 0.01 for histology), the number of reflux episodes lasting more than 5 min (p < 0.05 for endoscopy), and the frequency-duration index of reflux episodes (p < 0.001 for endoscopy, p < 0.01 for histology). Furthermore, strength reduction of peristalsis (< 60 mm Hg x s) was associated with acid exposure greater than 40%, in esophagitis patients. We conclude that the severity of reflux esophagitis, not only through endoscopy but also histologically, is related to the amount of reflux, as expressed by the duration and frequency of the reflux episodes. A very high amount of reflux is in turn associated with impairment of the esophageal body motility, as expressed by the amplitude and strength of esophageal body peristalsis. PMID- 8237939 TI - A multicenter, multiyear study of the safety and clinical utility of esophagogastroduodenoscopy in 20 consecutive pregnant females with follow-up of fetal outcome. AB - OBJECTIVES: To analyze the risks versus benefits of panendoscopy to the pregnant female and fetus. METHODS: Retrospective study of 20 consecutive pregnant patients admitted to three university teaching hospitals during 7 1/2 yr who underwent panendoscopy. RESULTS: Indications for panendoscopy included hematemesis in eight, vomiting and abdominal pain in five, vomiting in four, abdominal pain in two, and melena without hematemesis in one. Six patients were in the first trimester of pregnancy, eight were in the second trimester, and six were in the third trimester. Fourteen patients (70%) had a lesion diagnosed by panendoscopy, including esophagitis in seven, duodenal ulcer in two, gastritis in three, and Mallory-Weiss tear in two. In particular, all nine patients (100%) with gastrointestinal bleeding had a lesion identified by esophagogastroduodenoscopy. No significant endoscopic complications occurred. Panendoscopy did not induce labor in any patient. Fetal outcome was ascertained in 19 (95%) of the pregnancies. Seventeen infants were delivered at full term. Two were delivered at 33 and 35 weeks of gestation. No infant had a congenital malformation noted in the neonatal nursery. The mean infant Apgar scores were 8.2 +/- 1.3 (SD) at 1 min, and 9.1 +/- 0.3 (SD) at 5 min. CONCLUSIONS: In this study of 20 pregnancies, panendoscopy did not induce labor or result in congenital malformations. Panendoscopy is not absolutely contraindicated during pregnancy. Panendoscopy appears to be beneficial in medically stable pregnant patients with significant gastrointestinal bleeding. Panendoscopy should be performed with monitoring by electrocardiography and pulse oximetry after stabilization of vital signs, which may require transfusion of blood products and supplemental oxygen administration. PMID- 8237940 TI - Helicobacter pylori is a risk factor for hepatic encephalopathy in acute alcoholic hepatitis: the ammonia hypothesis revisited. The Veterans Administration Cooperative Study Group No. 275. AB - OBJECTIVE: To determine whether infection with Helicobacter pylori is a risk factor for portosystemic encephalopathy in patients with acute, moderate or severe alcoholic hepatitis. DESIGN: Prospective, multicenter cohort study. SETTING: Eight Veterans Affairs Hospitals. PATIENTS: A cohort of 273 male patients enrolled in a Department of Veterans Affairs Cooperative Study performed to evaluate the efficacy of oxandrolone in combination with nutritional supplementation in moderate or severe alcoholic hepatitis. MEASUREMENTS: Admission serum IgG antibody titers against H. pylori by a specific and sensitive ELISA, demographic characteristics of patients, degree of protein calorie malnutrition, presence of ascites, bilirubin level, and known risk factors for hepatic encephalopathy (gastrointestinal bleeding, azotemia, hepatorenal syndrome, infection, and severity of disease); outcome was the presence of portosystemic encephalopathy. RESULTS: Of 188 patients with decompensated alcoholic hepatitis available for analysis, 117 (62.2%) had encephalopathy. Ninety-two (78.6%) of these were infected with H. pylori, compared with 62% of patients without encephalopathy (p = 0.013). In a step-wise regression model, H. pylori was an independent risk factor (relative risk: 2.4, 95% CI: 1.2-4.8) adjusting for ascites and protein-calorie malnutrition. CONCLUSIONS: Patients with acute, moderate or severe alcoholic hepatitis have a high H. pylori infection rate (as determined by serology), and those infected are at higher risk for portosystemic encephalopathy. PMID- 8237941 TI - Pyogenic liver abscess in Taiwan: emphasis on gas-forming liver abscess in diabetics. AB - Ninety-seven cases of pyogenic liver abscesses in a 4-yr period were studied: 27.8% (27 cases) were associated with biliary tract stone, 5.2% (five cases) were associated with biliary tract cancer, and there were two cases of diabetes (2.1%) associated with anal infection, but 63.9% (63 cases) were diagnosed as cryptogenic. Forty patients (64.5%) in the cryptogenic group had diabetes mellitus, and 23 of them (23/40, 57.5%) had gas-forming infection. All patients received parenteral antibiotics therapy, percutaneous aspiration, drainage, or operation. The overall mortality was 16.5%. Diabetes mellitus alone, without demonstrable infectious foci, was an important predisposing factor for pyogenic liver infection. Furthermore, to evaluate the clinical importance of gas-forming pyogenic liver infections, we separated these 42 diabetic patients into gas forming and non-gas-forming groups, after sonography and CT scan. Klebsiella pneumoniae was the major pathogen in both groups. There was no significant difference in the clinical manifestations, complication, bacterial culture, or laboratory data between these two groups, except that the AST level was higher in the gas-forming group. However, the gas-forming group had higher mortality rate (30.4% vs. 5.3%). Gas-forming liver abscesses were common among the diabetics. Early and adequate drainage for pyogenic liver abscesses with parenteral antibiotics are crucial in their management. PMID- 8237942 TI - p53 expression in precancerous gastric lesions: an immunohistochemical study of PAb 1801 monoclonal antibody on adenomatous and hyperplastic gastric polyps. AB - To determine the chronology of p53 mutation in the gastric carcinogenic sequence, we studied p53 overexpression in premalignant lesions, including 17 adenomatous polyps (10/17 surrounded by intestinal metaplasia and 11/17 harboring foci of adenocarcinoma or severe dysplasia), and 18 hyperplastic polyps (4/18 with focal adenomatous changes). Immunohistochemistry with PAb 1801 monoclonal antibody was performed on archival material; p53 nuclear staining was seen in 10/17 adenomas, but was limited to the foci of adenocarcinoma in three cases. Five adenomas with foci of severe dysplasia or carcinoma were nonreactive. Intestinal metaplasia, normal gastric mucosa, and 14/18 hyperplastic polyps were nonreactive. p53 Reactivity observed in four hyperplastic polyps was limited to adenomatous foci. These results suggest that p53 overexpression occurs in dysplastic epithelium of precancerous gastric lesions. Its absence in chronic atrophic gastritis with intestinal metaplasia suggests it is a relatively late event in the gastric carcinogenic sequence. PMID- 8237943 TI - Expression of plasminogen activators and their inhibitors in human pancreatic carcinoma: immunohistochemical study. AB - Expression of plasminogen activators (PA), tissue type (t-PA) and urokinase type (u-PA), as well as PA inhibitors (PAI), type-1 (PAI-1) and type 2 (PAI-2), were investigated immunohistochemically in 97 human pancreatic carcinomas. u-PA expression predominated in pancreatic carcinomas, compared with t-PA [u-PA expression in 76 specimens (78.4%) and t-PA in eight specimens (8.2%)]. PAI-1 expression was detected in 80 carcinoma specimens (82.5%) and PAI-2 in 79 carcinoma specimens (81.4%). PAI-2 expression was significantly lower in carcinomas with peritoneal metastasis (p < 0.02). Strong PAI-2 expression was associated with significantly higher survival than negative or weak PAI-2 expression (p < 0.05). We conclude that immunohistochemical analysis of PAI-2 expression in pancreatic carcinomas may yield important prognostic information. PMID- 8237944 TI - Barrett's esophagus in children with cystic fibrosis: not a coincidental association. AB - Barrett's esophagus (BE) is a premalignant condition, and a recognized complication of severe gastroesophageal (GE) reflux. Children with cystic fibrosis (CF) have a marked predilection to develop GE reflux, but Barrett's esophagus is one complication of GE reflux not previously described in CF. We describe in detail two adolescents with CF who were found to have Barrett's esophagus, and mention three other cases. The presence of Barrett's esophagus in CF patients may be missed because GE reflux is often relatively silent in CF, because patients may consider mild upper gastrointestinal (GI) symptoms as "part of CF," and because of the nature of Barrett's epithelium itself. Upper gastrointestinal (GI) endoscopy with documentation of landmarks and multiple targeted biopsies should be performed in children with CF with even mild symptoms of GE reflux or an abnormal 24 h intra-esophageal pH study. Any biopsies containing columnar epithelium should be stained with Alcian blue at pH 2.5 to look for goblet cell metaplasia, i.e., Barrett's esophagus. Children with CF may be a high-risk group for development of Barrett's esophagus and its complications, especially given the increased survival in CF. PMID- 8237945 TI - Dieulafoy-like lesion of the rectum presenting with exsanguinating hemorrhage: successful endoscopic sclerotherapy. AB - The classic Dieulafoy lesion is a minute gastric mucosal defect that bleeds massively from an exposed artery. Similar lesions have been identified in the small intestine, colon, and rectum. In this report, we describe a patient who presented with an exsanguinating hemorrhage from a small rectal ulcer with a visible vessel. Control of the bleeding was achieved with endoscopic sclerotherapy using absolute alcohol and 1.5% sodium tetradocyl sulfate, with eventual complete healing of the site. PMID- 8237946 TI - Vascular anatomy of duodenal varices: angiographic and histopathological assessments. AB - Duodenal varices are a rare occurrence, and the diagnosis and control of related bleeding is problematic. The vascular anatomy of duodenal varices in four patients was examined angiographically. The incidence of these varices was 0.4% in the patients with portal hypertension seen in our clinics from 1982 to 1991. The transparent specimen obtained at autopsy and treated with methyl salicylate and portographies in the remaining three patients revealed that the duodenal varix consisted of a single vessel with afferent and efferent vessels, forming a portasystemic shunt in the retroperitoneum. The varix transversed the duodenum and was present in the submucosal layer of the posterior wall. The afferent vessel was the superior or inferior pancreaticoduodenal vein originating in the portal vein trunk or superior mesenteric vein. The efferent vein drained into the inferior vena cava. In view of these observations, devascularization or percutaneous transhepatic obliteration of the varix would be our first choice of treatment for patients with duodenal varices. PMID- 8237947 TI - Ultrasound-assisted percutaneous endoscopic gastrostomy in a patient with advanced ovarian carcinoma and recurrent intestinal obstruction. AB - We report a case of recurrent bowel obstruction in a 56-yr-old woman secondary to progressive ovarian carcinoma. Due to a history of multiple prior surgeries, intraperitoneal chemotherapy, radiotherapy, and progressive intra-abdominal disease, placement of a percutaneous endoscopic gastrostomy (PEG) for decompression was thought to carry significant risk if standard techniques were used. We therefore used transcutaneous ultrasound imaging at the time of endoscopy, which allowed for rapid localization of an appropriate area for PEG placement, thereby minimizing the risk of inadvertently entering adjacent organs or metastatic foci. To our knowledge, this is the first such case in which ultrasound-assisted PEG placement has been utilized. PMID- 8237948 TI - Small, polypoid-appearing carcinoid tumors of the rectum: clinicopathologic study of 16 cases and effectiveness of endoscopic treatment. AB - Fifteen cases of small-size rectal carcinoid tumors (RCTs) were discovered among a total of 21,522 healthy teachers who received proctosigmoidoscopy. Adding another case seen in the outpatients, a total of 16 RCTs were clinicopathologically studied. The average age of the patients was 48.8 yr, and the incidence was predominantly among males. Prior to endoscopy, digital examination of the rectum revealed a palpable firm nodule in 13 patients. On endoscopy, RCTs were generally round, yellow-discolored polyps with a size less than 13 mm in diameter, covered by a normal-appearing mucosa. An erythematous change or depression was seen in three tumors. Histologically, they showed pure insular, trabecular, or mixed structures, and tumor cells were confined to the mucosa and/or submucosa in all cases. Immunohistochemically, RCTs never failed to show focal or diffuse positivities for chromogranin A and/or neuron-specific enolase. Pancreatic polypeptide was immunostained in 14 tumors to a varying degree. The tumors having been safely treated by endoscopic polypectomy and/or surgical excision, all of the patients are alive and clinically free of disease during the average observation period of 79 months. PMID- 8237949 TI - Radiologic regression of primary sclerosing cholangitis following combination therapy with an endoprosthesis and ursodeoxycholic acid. AB - Bile duct strictures seen in patients with primary sclerosing cholangitis are generally considered irreversible. Oral ursodeoxycholic acid therapy has been shown to improve symptoms and biochemical abnormalities, as well as reverse intrahepatic alterations in patients with biliary strictures. Endoscopic therapy also has been effective in relieving symptoms, improving biochemical and radiologic abnormalities, and preventing recurrent cholangitis in patients with primary sclerosing cholangitis. We report a case in which a combination of prolonged endoscopic stenting and oral bile acid therapy resulted in regression of cholangiographic changes and produced a sustained clinical remission. PMID- 8237950 TI - Achalasia of the esophagus in secondary amyloidosis. PMID- 8237951 TI - Cholestatic hepatitis associated with the use of nafcillin. PMID- 8237952 TI - A pseudocyst infected by Torulopsis glabrata: a unique problem. PMID- 8237953 TI - Gastric plasmacytoma: a rare cause of massive gastrointestinal bleeding. PMID- 8237954 TI - A case of splenic penetration by a benign gastric ulcer. PMID- 8237955 TI - Acute appendicitis secondary to metastatic cholangiocarcinoma. PMID- 8237956 TI - A case of small combined hepatocellular and cholangiocellular carcinoma arising in a nodule of atypical adenomatous hyperplasia of the liver. PMID- 8237957 TI - Natural history of post-transfusion hepatitis: the wolf in sheep's clothing. PMID- 8237958 TI - Hepatitis C: you cannot blame it on your mother. PMID- 8237959 TI - Acid inhibition and amoxicillin activity against Helicobacter pylori. PMID- 8237960 TI - Primary biliary cirrhosis and inflammatory bowel disease. PMID- 8237961 TI - Intestinal permeability to 51Cr-ethylenediaminetetraacetate in patients with ulcerative colitis. PMID- 8237962 TI - Finger clubbing in gastrointestinal polyposis. PMID- 8237963 TI - Combined intravenous and oral omeprazole/amoxicillin therapy in patients with Helicobacter pylori-positive acute peptic ulcer bleeding. PMID- 8237964 TI - Embryological development of the duodenal papilla, and related diseases: primitive ampulla theory. PMID- 8237965 TI - Lipoma of the colon: self amputation. PMID- 8237966 TI - Miscarriages among female physical therapists who report using radio- and microwave-frequency electromagnetic radiation. AB - Physical therapists are exposed to radio- and microwave-frequency electromagnetic radiation by operating shortwave and microwave diathermy units. Recent studies suggest that use of shortwave diathermy is associated with an excess risk of birth defects, perinatal deaths, and late spontaneous abortions among the offspring of exposed female therapists. To assess the impact of occupational use of microwave and shortwave diathermy at the time of conception, the authors mailed questionnaires to 42,403 physical therapists in 1989. Both occupational and reproductive histories were obtained. Exposures to shortwave and microwave diathermy were both assessed in the same fashion and were examined in relation to early recognized fetal loss in a nested case-control design. A total of 1,753 case pregnancies (miscarriages) were matched to 1,753 incidence density control pregnancies (other pregnancies except ectopic pregnancies). A pregnancy was considered "exposed" if the mother reported using microwave or shortwave diathermy anytime during the 6 months prior to the first trimester or during the first trimester. Pregnancies of mothers reporting microwave use 6 months prior to the pregnancy or during the first trimester were more likely to result in miscarriage (odds ratio (OR) = 1.28, 95% confidence interval (CI) 1.02-1.59). The odds ratio increased with increasing level of exposure (chi 2 = 7.25, p < 0.005). The odds ratio in the highest exposure group (20 or more exposures/month) was 1.59. The overall odds ratio was slightly lower after it was controlled for prior fetal loss (OR = 1.26, 95% CI 1.00-1.59), but the exposure-response effect remained (chi 2 = 5.17, p < 0.01). The risk of miscarriage was not associated with reported use of shortwave diathermy equipment (OR = 1.07, 95% CI 0.91-1.24). The odds ratio in the highest exposure group was 0.87. PMID- 8237967 TI - Exposure-response analysis of cancer mortality in a cohort of workers exposed to ethylene oxide. AB - The authors previously reported results from the largest cohort mortality study of ethylene oxide-exposed workers that has been conducted to date. Here they extend their previous work by quantitatively examining the relation between cancer mortality and ethylene oxide exposure. This study included workers from 13 of the 14 geographically distinct facilities that were included in the previous investigation. These facilities began regularly using ethylene oxide to sterilize medical supplies or spices sometime between 1938 and 1969. Workers were followed from first exposure through December 31, 1987. Historical exposures to ethylene oxide were estimated using a regression model. Standard life-table analysis was used to examine cancer mortality in three categories of cumulative exposure to ethylene oxide. The Cox proportional hazards model was also used to examine cumulative and other measures of ethylene oxide exposure as predictors of cancer mortality. In both the life-table analysis and the Cox model, a positive trend was observed in all lymphatic and hematopoietic cancer mortality for cumulative ethylene oxide exposure. This trend was strengthened when ethylene oxide exposures 10 years prior to death were discounted (lagged) and when the analysis was restricted to neoplasms of lymphoid cell origin. Despite limitations discussed in this paper, the authors believe that these findings provide some support for the hypothesis that exposure to ethylene oxide increases the risk of mortality from lymphatic and hematopoietic neoplasms. The authors intend to continue follow-up of this relatively young cohort, which may allow more definitive conclusions to be drawn in the future. PMID- 8237968 TI - Association of body fat distribution and family histories of breast and ovarian cancer with risk of postmenopausal breast cancer. AB - The authors recently published data from a prospective cohort study of postmenopausal women (N Engl J Med 1992;326:1323-9) which suggested that a high waist/hip ratio was associated with a significantly increased risk of breast cancer in women with a positive family history of breast cancer. Since families with aggregations of breast and ovarian cancer demonstrate more consistent genetic linkage to markers on chromosome 17q than do families with breast cancer alone, the authors performed additional analyses to examine whether the previously observed associations with waist/hip ratio differed when family history of breast cancer was partitioned according to whether or not ovarian cancer was also present in a close relative. Between 1986 and 1990, 620 incident cases of breast cancer were identified in a cohort of 37,105 postmenopausal Iowa women. A family history of breast cancer in first-degree relatives was associated with a relative risk of 1.34 (95% confidence interval (CI) 1.07-1.69); a family history of both breast cancer and ovarian cancer was associated with a relative risk of 2.36 (95% CI 1.12-4.98). Consistent with the authors' findings after 4 years of follow-up, a high waist/hip ratio (> or = 80th percentile vs. < 80th percentile) was associated with increased risk of breast cancer in the presence of a family history of breast cancer (relative risk (RR) = 2.10, 95% CI 1.43 3.09) but not in the absence of a family history of breast cancer (RR = 1.12). The combination of a high waist/hip ratio with a family history of breast and ovarian cancer was associated with 4.83-fold increased risk (95% CI 1.55-15.1). Neither a family history of breast cancer nor a family history of ovarian cancer was associated with significantly increased risk in the absence of a high waist/hip ratio. PMID- 8237969 TI - Randomized trial of the effect of calcium supplementation on fecal risk factors for colorectal cancer. AB - Newmark et al. (J Natl Cancer Inst 1984; 72:1323-8) hypothesized that supplementation with calcium would decrease the concentration of bile acids in aqueous phase feces and that such a reduction would reduce the risk of malignant disease in the bowel. A randomized trial was therefore undertaken to examine the effects of calcium supplementation on fecal biochemistry. A total of 68 men between 40 and 60 years of age volunteered to participate after having been selected randomly from population lists in Scarborough, a city in the Toronto area, Ontario, Canada. Participants were randomly assigned to receive a daily supplement of 3 g of calcium carbonate or a sucrose placebo for a period of 1 week. Fecal samples were collected for 2 days prior to supplementation and for the last 2 days of supplementation. Records of all foods consumed were kept throughout the study period. The average concentration of total soluble bile acids fell in the placebo group (-11.2 g/ml) but increased slightly in the calcium group (1.4 g/ml). Similar patterns of change were observed for deoxycholic acid (placebo, -3.0 g/ml; calcium, 4.5 g/ml). The distribution of changes in total bile acids and deoxycholic acid differed between randomization groups at the 10 percent level of significance in univariate analysis. After adjustment for initial stool chemistry and initial levels and changes in nutrient intake, no reduction in fecal bile acid levels was observed in association with calcium supplementation. In fact, a statistically significant (p = 0.05) increase in deoxycholic acid concentration remained in the calcium supplemental group. Thus, this study was unable to support the hypothesis that calcium supplementation alters aqueous phase bile acids in the manner hypothesized to be consistent with protection from colorectal cancer. PMID- 8237970 TI - Relation between blood lead and urinary biogenic amines in community-exposed men. AB - The cross-sectional relation between levels of urinary biogenic amines (dopamine, epinephrine, norepinephrine, and serotonin) and levels of blood lead was examined in a study of 645 male participants from a longitudinal study of aging. This stable population of men had initially been recruited from communities in and around Boston, Massachusetts, and had not been selected with regard to lead exposure. Blood lead samples and 24-hour and 2-hour urine specimens were collected during regularly scheduled clinic visits. In multivariate linear regression step-forward models, 24-hour epinephrine excretion was significantly and positively associated with blood lead (beta = 0.101 microgram(microgram/dl)-1 blood lead, SE (standard error) (beta) = 0.045, p = 0.026). Twenty-four-hour norepinephrine excretion was positively associated with blood lead (beta = 0.023 microgram(microgram/dl)-1 blood lead, SE(beta) = 0.029, p = 0.425), and both 24 hour dopamine (beta = -4.35 microgram(microgram/dl)-1 blood lead, SE(beta) = 6.90, p = 0.529) and 2-hour serotonin (beta = -0.348 microgram(microgram/dl)-1 blood lead, SE(beta) = 0.277, p = 0.210) excretion were negatively associated with blood lead; however, these relations did not achieve statistical significance. An increase of 10 micrograms/dl in blood lead was associated with an increase in epinephrine excretion of 11 micrograms/24 hours. These results support the hypothesis that epinephrine metabolism is influenced by low levels of lead exposure. PMID- 8237971 TI - Determinants of incident non-insulin-dependent diabetes mellitus among blacks and whites in a national sample. The NHANES I Epidemiologic Follow-up Study. AB - The excess incidence of non-insulin-dependent diabetes mellitus noted among African Americans in the past two decades may be attributable to variations in the distribution of specific risk factors, or the impact of these risk factors may differ by ethnicity or sex. Over the 16 years (1971-1987) of the First National Health and Nutrition Examination Survey (NHANES I) Epidemiologic Follow up Study, 880 incident cases of diabetes mellitus developed among 11,097 white and black participants who were between the ages of 25 and 70 years at baseline. There were substantial differences among the four race/sex groups with respect to age at baseline, as well as marked differences in the distribution of several major risk factors for diabetes, including obesity, subscapular and triceps skinfold thickness, blood pressure, income, activity, and educational level. The age-adjusted incidence of diabetes over the course of the study was 15.0% among black women, while it was 10.9% among black men. White women and men experienced similar, more moderate risks of 7.0% and 6.9%, respectively. The 100% excess risk among black women and the 50% excess among black men can in large measure explain the recent marked increase in diabetes rates in the black community. Furthermore, at nearly every level of obesity, blacks had a higher risk of diabetes than whites, suggesting that other factors contributed to risk. A significant interaction between race and body mass index (weight (kg)/height(m)2) was likewise demonstrated in multivariate analysis. Baseline age, race, body mass index, and ratio of subscapular skinfold to triceps skinfold were significantly related to incident diabetes, both overall and in separate models for men and women; in the entire cohort and in women alone, blood pressure, activity level, and education also contributed to risk. Other interactions were tested but were not found to be important. Despite sampling difficulties and inconsistencies in the data, the NHANES I Epidemiologic Follow-up Study provides evidence that the associations of anthropometric and sociodemographic variables with diabetes may vary among subgroups which have different mean levels and distributions of these risk factors. PMID- 8237972 TI - Self-rated health, mortality, and chronic diseases in elderly men. The Zutphen Study, 1985-1990. AB - The value of self-rated health in predicting mortality and the incidence of chronic diseases was studied in a cohort of 783 elderly Dutch men in the Zutphen Study. In 1985, 48% of the men felt "healthy," while 12% felt "moderately healthy" or "not healthy." As of 1990, 23% of the 783 men had died. Survival analysis showed that self-rated health was highly predictive of subsequent 5-year mortality from all causes (p < 0.001). When adjusted for the presence of major chronic diseases, age, medication use, smoking, alcohol consumption, physical activity, body mass index, systolic blood pressure, serum cholesterol, education, marital status, and family history of chronic diseases, the relative risk for "moderately healthy" or "not healthy" men compared with "healthy" men was 2.7 (95% confidence interval (CI) 1.8-4.3). Analysis of cause-specific mortality revealed that self-rated health was associated with cardiovascular mortality (crude relative risk (RR) = 2.7), but this finding resulted mainly from confounding by baseline prevalence of cardiovascular diseases (adjusted RR = 1.9, 95% CI 0.9-3.8). However, self-rated health was an independent risk factor for cancer mortality (adjusted RR = 4.2, 95% CI 1.9-9.4) and mortality due to other causes (adjusted RR = 3.0, 95% CI 1.2-7.8). Self-rated health did not independently predict the incidence of chronic diseases. This suggests that self rated health especially affects fatality from chronic diseases rather than their onset, and this issue should be pursued further. PMID- 8237973 TI - Epidemiologic studies of Escherichia coli diarrheal infections in a low socioeconomic level peri-urban community in Santiago, Chile. AB - The incidence of diarrhea due to six categories of diarrheogenic Escherichia coli was determined in two pediatric cohorts in a low socioeconomic level community in Santiago, Chile, with access to chlorinated water. An age cross-sectional cohort of 340 children aged birth to 47 months was assembled. A newborn cohort was assembled by enrolling 10-12 newborns monthly for 12 months. Episodes of diarrhea were detected by twice weekly household visits. E. coli from stool cultures of cases and matched controls were hybridized with DNA probes specific for enterotoxigenic, enteroinvasive, enteropathogenic, enterohemorrhagic, enteroaggregative, and diffuse adherence E. coli. Overall, the incidence of diarrhea was low (2.1 episodes/infant/year). Nevertheless, a putative E. coli enteropathogen was found in a large proportion of diarrheal episodes, particularly during the summer. In both cohorts, enterotoxigenic E. coli were important pathogens. Enteropathogenic E. coli were incriminated during the first year of life in the newborn cohort, where they were found significantly more often in cases (p = 0.021) than in controls; beyond this age, isolation rates were similar. In contrast, the relative risk of isolation of diffuse adherence E. coli increased with age in the age cross-sectional cohort, where, overall, the difference in rate of isolation between cases and controls was significant (p = 0.0024). Enteroinvasive and enterohemorrhagic E. coli were isolated infrequently. Enteroaggregative E. coli were encountered equally in cases and controls. Facile transmission of E. coli enteropathogens is occurring in this community despite the availability of potable water. PMID- 8237974 TI - Re: "Arsenic ingestion and internal cancers: a review". PMID- 8237975 TI - Re: "Declining seroprevalence and transmission of HTLV-I in Japanese families who emigrated to Hawaii". PMID- 8237976 TI - Re: "Magnetic fields of video display terminals and spontaneous abortion". PMID- 8237977 TI - Re: "Autoimmunity and genetics contribute to the risk of insulin-dependent diabetes mellitus in families: islet cell antibodies and HLA DQ heterodimers". PMID- 8237978 TI - Outbreak of Legionnaire's disease linked to a decorative fountain by molecular epidemiology. AB - The incubation period of Legionnaires' disease in five patients was traced to attendance at conventions in a hotel in the Orlando, Florida, area between January 6 and February 2, 1992. The five case patients (mean age, 69 years) were older than 55 randomly chosen controls (mean age, 53 years) who had also attended one of the same conventions (p = 0.007). All case patients were males, as were 40% of the controls (p = 0.01). No significant differences in exposures were found between case patients and controls, but all case patients and 65% of the controls reported exposure to a decorative fountain in the hotel lobby. Water from the fountain was the only one of 55 environmental specimens to test positive for Legionella. Both the environmental isolate and the only clinical isolate were Legionella pneumophila serogroup 1, with identical patterns identified on monoclonal antibody subtyping and pulsed-field gel electrophoresis (PFGE) of genomic restriction fragments. The fountain's recirculating system had been irregularly maintained, and water in the fountain may have been heated by submersed lighting. These findings demonstrate the utility of monoclonal antibody subtyping and PFGE of genomic restriction fragments in assessing the significance of environmental isolates of L. pneumophila, especially when other epidemiologic findings are inconclusive. They also show that decorative fountains may be a potential source of infection with L. pneumophila, and emphasize the need for standard maintenance and disinfection procedures. PMID- 8237979 TI - Malaria and perinatal mortality in central Sudan. AB - Hospital and community studies were conducted in Central Sudan during 1989 and 1990 to determine the association between maternal malaria and perinatal mortality. There were 197 cases of stillbirth and 812 controls in the hospital study. In the community study, 36 perinatal and 31 neonatal deaths were compared with 1,505 and 1,495 survivors of the early neonatal and entire neonatal periods, respectively. There was no overall association between perinatal mortality and malaria. However, the risk of stillbirth (particularly macerated stillbirth) was significantly increased among women who reported malaria attacks in the first and second trimesters of pregnancy (odds ratio (OR) = 1.4, 95% confidence interval (CI) 1.1-1.9). A reduced risk was associated with attacks in the third trimester (OR = 0.4, 95% CI 0.2-0.8), but this probably reflects a bias resulting from shorter gestation in cases of stillbirth. Increased risk of neonatal mortality was associated with maternal malaria (OR = 2.1, 95% CI 1.0-4.5). In areas where malaria is prevalent, it is recommended that malaria prevention by personal protection, prophylaxis, and treatment be initiated early in pregnancy. PMID- 8237980 TI - Self-reported morbidity and disability among Trappist and Benedictine monks. AB - Studies of religious groups that impose prudent lifestyles on their members show low mortality rates in these groups, but it is unclear whether their morbidity rates are also low. The authors studied the prevalence of self-reported morbidity and disability among Trappist and Benedictine monks in the Netherlands. A health interview survey was administered in seven monasteries; the response rate was 67% (n = 134). Rates of morbidity and disability among monks were compared with those among all Dutch males by calculation of standardized morbidity ratios, adjusting for age and (in the case of morbidity) education. Self-reported morbidity among monks was similar to that in the general population (e.g., for one or more chronic conditions: standardized morbidity ratio = 1.07, 95% confidence interval 0.89-1.26), but rates of disability related to activities of daily living were much higher (e.g., for any trouble sitting down and getting up from a chair: standardized morbidity ratio = 2.21, 95% confidence interval 1.44-3.32). The authors hypothesize that a prudent lifestyle may prolong life, but at the expense of a higher prevalence of disability. PMID- 8237981 TI - Effectiveness of the Waksberg telephone sampling method for the selection of population controls. AB - The effectiveness of the Waksberg telephone sampling method and practical aspects of its implementation were evaluated during the selection of control subjects for a population-based case-control study conducted in Maryland, Virginia, West Virginia, and Washington, DC, in 1991. The first stage of the two-stage sampling procedure was equivalent to simple random sampling. Each telephone number selected was dialed up to 13 times to establish the type of connection. Residential connections were used to generate additional telephone numbers in the second stage by changing the last two digits. Overall, 3,860 telephone numbers yielded 1,311 residences. The proportion of residences was higher in the second stage (54%) than in the first (11%), while that of numbers "not in service" was lower (12% vs. 67%, respectively). Fewer calls (8,735) were made than would have been required by a simple random sampling procedure (up to 18,522). The geographic distribution of residences was similar at both stages of sampling. At the end of enrollment, each sampling unit had yielded 9.8 residences, but only 64% reached 9-11 residences. Compliance rates were 96% for screening (1,259 of 1,311 residences), 90% for interviewing (362 of 403), and 86% overall. Relative to simple random sampling, the Waksberg method considerably reduced the total number of telephone calls, but the sampling process was more complex to manage. This method is particularly suitable for selecting population controls when the density of residential connections is low and variables of interest are not homogeneous within sampling units. Random digit dialing remains an effective method for selecting population controls. PMID- 8237982 TI - Family history, age, and risk of fatal breast cancer. AB - A family history of breast cancer potentially identifies women at high risk for whom mammography may be especially valuable. The authors examined the association of fatal breast cancer and family history in a large, prospective study of US adults. After 6 years of follow-up, 880 cases of fatal breast cancer were observed in a cohort of 604,412 women who were cancer free at interview in 1982. Cox proportional hazards modeling found that a family history of breast cancer in a mother or sister was significantly related to fatal breast cancer risk (rate ratio = 1.59, 95% confidence interval (CI) 1.29-1.95). The association was greatly modified by age; the rate ratio was 4.91 (95% CI 1.39-17.3) in women aged < 40 years at enrollment compared with 1.28 (95% CI 0.78-2.11) in women aged > or = 70 years. Among young (aged < 50 years) women, the risk associated with a positive family history was greatest if the relative was also young at the time of diagnosis (rate ratio = 5.22, 95% CI 3.00-9.11). These associations were not altered in multivariate analyses controlling for known breast cancer risk factors. In the authors' data, family history is strongly predictive of early fatal breast cancer, particularly when the disease is diagnosed before age 50 in the relative. Women with a family history of early breast cancer should receive special instructions regarding mammography and early detection. PMID- 8237983 TI - Incidence of cancer among lighthouse keepers exposed to asbestos in drinking water. AB - The study population comprises 690 Norwegian male lighthouse keepers whose water supply came from cisterns that received rain water off asbestos-cement-tiled roofs. The asbestos-cement tiles were installed in the late 1950s, and two decades later the fiber content in the tap water was analyzed. The fiber content ranged from 1,760 to 71,350 million fibers per liter, which is significantly higher than measured in any other Norwegian public water supply. During the follow-up period, 1960-1991, no statistically significant excess risk was found for any type of cancer in the group with a latency period of 20 years or more, except for stomach cancer (11 observed cases vs. 4.57 expected, standardized incidence ratio = 241, 95% confidence interval 120-431). No cases of malignant mesothelioma were found. The study is limited by lack of knowledge as to when the tiles began to deteriorate and, thus, the magnitude of total exposure as well as by the inability to control for such potential confounding factors as diet. PMID- 8237984 TI - Relation of age, exercise, anthropometric measurements, and diet with glucose and insulin levels in a population aged 70 years and over. AB - A community-based sample of people > or = 70 years from Mosgiel, New Zealand, was investigated to determine the relation of age, diet, exercise, drugs, and anthropometric measurements with glucose and insulin. From an initial sample of 856 subjects on August 1, 1988, 782 (91.4%) completed the questionnaires and physical examination. Glucose was estimated in 726 subjects and insulin in 607 subjects 2 hours after a standardized meal. In the multivariate analysis for women, glucose was related to age and exercise, and insulin was related to glucose levels, triceps skinfold thickness, and waist/hip ratios. In the multivariate analysis for men, none of the variables was related to glucose levels; insulin levels were related to glucose and waist/hip ratio. Impaired glucose tolerance in women was associated with high body mass index, waist/hip ratio, lower exercise levels, and the taking of thiazide drugs or oral steroids. In men, no significant model to identify those with impaired glucose tolerance could be developed. Glucose and insulin levels in women were related to age and external factors, particularly exercise and anthropometric measurements. In men, external factors were less clearly related to glucose and insulin levels, but this lack of association in men may be due to the smaller number of men in the sample, their younger age, and the narrower range of values found in the men. PMID- 8237985 TI - Comparison of recalled and validated oral contraceptive histories. AB - From a case-control study of the relation between oral contraceptives and breast cancer carried out in East Germany during 1982-1986, the authors obtained information on oral contraceptive use through interviews of study subjects and from the records of prescribing gynecologists. The degree of agreement regarding information from these two sources was assessed for 234 breast cancer cases and 524 controls who had ever used oral contraceptives. Agreement between information obtained from medical records and that from interviews on total duration of use, number of episodes of use, and time since first and last use was reasonably good, and levels of agreement did not differ appreciably between cases and controls. Lower levels of agreement were observed for individual brand names and the duration of use of specific brands. Attempts should be made to obtain information on specific brands from medical records when investigating the effects of individual preparations. PMID- 8237986 TI - Randomized trial of use of a monetary incentive and a reminder card to increase the response rate to a mailed health survey. AB - Insufficient response rates are a frequent problem in mailed epidemiologic health surveys. As part of a health survey of 1,235 young adults conducted from November 1992 through January 1993 in Geneva, Switzerland, a randomized factorial trial was carried out to assess the contributions of two tactics to increase response rates. The first incentive was the promise of sending 10 Swiss francs (7 US dollars) to respondents; the second was a red postcard, mailed 2 days after the questionnaire, which reminded potential participants to complete the questionnaire. The most effective strategy according to interim analysis was to be selected for follow-up mailings. Two weeks after the first mailing, response rates were 65% for those who had received both incentives, 57% for those who had been offered the money reward only, 54% for recipients of the reminder card, and 48% for those who had received neither incentive (p based on chi 2 (3 df) test < 0.001). The relative hazards of responding were 1.34 (95% confidence interval 1.15-1.55) for the money reward and 1.22 (95% confidence interval 1.05-1.41) for the reminder card. Follow-up mailings to all nonrespondents included both incentives. At the end of data collection, 1,007 persons (82%) had returned the questionnaire. The final response rates were 83, 84, 82, and 78% in the four groups, respectively (p = 0.29). Both the promise of a monetary reward and a reminder postcard considerably improved early response rates. The use of both incentives in follow-up mailings brought final response rates above 80%, except for those who had received no initial incentive. Testing various combinations of incentives in the early stages of a mailed health survey may be a generally useful strategy. PMID- 8237987 TI - Use of surnames to identify individuals of Chinese ancestry. AB - The objectives of this study were to develop and test surname lists for identifying Chinese ancestry. The Ontario all-cause mortality database for the period 1982-1989 was randomly split into source and test data sets. Frequencies by birthplace were compiled for each surname in the source data set, by sex, and the surnames were weighted based on their positive likelihood ratios. Lists of Chinese surnames were then assembled based on varying cutoff levels, and screening performance indicators for each list were calculated, including sensitivity, specificity, positive and negative predictive values, post-test odds, positive likelihood ratio, and yield. The internally generated lists were evaluated in the test data set. Results indicated that surnames have a good potential to identify individuals of Chinese origin. In the source data set, at a cutoff level of 100 for males (217 surnames) and females (210 surnames), both sensitivity and the positive predictive value of the surname lists for males and females were very high, above 80%, and the positive likelihood ratio was above 600. In the test data set and using the same surname lists, the sensitivity, positive predictive value, and positive likelihood ratio remained at a high level: 73%, 81%, and 603, respectively, for males; and 73%, 84%, and 772, respectively, for females. Various scenarios and their methodological implications are discussed. PMID- 8237988 TI - Risk factors associated with cervical human papillomavirus infections: a case control study. AB - As a part of the long-term prospective follow-up study conducted for women with genital human papillomavirus (HPV) infections in Kuopio University Hospital, Finland, since 1981, a case-control study was designed to assess risk factors for genital HPV infections. The cases (n = 691) were women who had been invited to attend the follow-up program on the basis of an abnormal cervical smear consistent with HPV-induced cytopathic changes, i.e., had a clinical HPV infection. The controls (n = 706) were a randomly selected group of women who had normal smears in the screening. Both groups were asked to fill in an extensive questionnaire focusing on detailed epidemiologic data on previous gynecologic and obstetric history, sexual practices, sexual partners, and smoking habits. In the multivariate analysis, eight variables emerged as independent risk factors for prevalent HPV infection. These variables could explain over 80% of the risk for infection. The risk for the infection varied with age, being highest in the age group 20-29 years, thereafter declining in the following 10-year age groups. The strongest independent risk factor was the number of sexual partners during the past 2 years (adjusted odds ratio = 12.1; 95% confidence interval 4.3-33.8 for five or more vs. one or no partners). Among the independent risk factors that increased the risk were also current smoking (adjusted odds ratio = 2.7; 95% confidence interval 1.7-4.3), warts in sexual partner(s) (adjusted odds ratio = 3.2; 95% confidence interval 1.6-6.5), and increasing frequency of sexual intercourse per week. Independent risk factors with a protective effect included a normal result in the last Papanicolaou smear, regular use of an intrauterine device as a contraceptive method, and good personal hygiene. No significant association between oral contraceptive use and risk for HPV infection was found. Condom use did not result in protection from the infection. The results of this study support the concepts that sexual intercourse is the main form of transmission among adults and that sexual promiscuity is the most important determinant for genital HPV infections. PMID- 8237989 TI - Divergent mortality for male and female recipients of low-titer and high-titer measles vaccines in rural Senegal. AB - The female/male mortality ratio among unimmunized children and children vaccinated with standard or high-titer measles vaccines was examined for all children born in the period 1985-1991 in a rural area of Senegal. The female/male mortality ratio from 9 months to 5 years of age for unvaccinated children was 0.94 (95% confidence interval (CI) 0.75-1.19), significantly different from the ratio of 0.64 (95% CI 0.48-0.85) for recipients of the Schwarz standard measles vaccine (p = 0.040). In the 4-year period, where high-titer measles vaccines were used in the study area, the female/male mortality ratio was 1.33 (95% CI 1.00 1.78) for recipients of high-titer Edmonston-Zagreb or Schwarz vaccines compared with 0.67 (95% CI 0.42-1.07) for recipients of the Schwarz standard vaccine (p = 0.013). Hence, the Schwarz standard and high-titer measles vaccines have divergent sex-specific effects on mortality throughout childhood. Further studies of the underlying mechanisms are needed. PMID- 8237990 TI - Results of a rubella screening program for hospital employees: a five-year review (1986-1990). AB - The Barnes Hospital Employee Health Service (St. Louis, Missouri) rubella screening program was evaluated over the 5-year period between January 1, 1986, and December 31, 1990. A total of 6,969 new employees were hired, and 6,115 (87.7%) were screened for evidence of rubella immunity by the Employee Health Service. Rubella serology was performed on 5,893 (96.4%) of the screened employees, while 222 (3.6%) had documentation of prior rubella vaccination or rubella infection. The absence of immunity was identified in 325 employees or 5.3% of all those screened. Women were more frequently screened by the Employee Health Service than were men (p < 0.0001), and blacks were more frequently screened than were non-Hispanic Caucasians (p < 0.0001). Physicians were less frequently screened than were other departmental groups (p < 0.0001). The rate of seronegativity for each year of hire varied from 4.45 to 6.76%, but these differences were not significant. Logistic regression analysis demonstrated that 5-year birth cohorts correlated significantly with serologic status. Employees born in 1960-1964 were least likely to be seronegative, and employees born in 1970 or later were most likely to be seronegative. Sex, race, and department group were not predictive of serologic status, although significant differences in results from different rubella assays were detected. Only 13.8% of seronegative employees were subsequently vaccinated by the Employee Health Service. This study demonstrates a lower seronegativity rate than did previous studies. It identifies groups of employees likely to escape rubella screening and low vaccination rates. It finds increasing seronegativity among employees born after 1964 that correlates with the reported increasing rates of rubella in the United States. PMID- 8237991 TI - Re: "Lymphohematopoietic cancer in styrene-butadiene polymerization workers". PMID- 8237992 TI - Recombinant human granulocyte-macrophage colony-stimulating factor plus recombinant human erythropoietin may improve anemia in selected patients with myelodysplastic syndromes. AB - The purpose of this study was to improve erythropoiesis in patients with anemia due to myelodysplastic syndromes (MDS). We treated 13 patients first with recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) for 6 weeks, then with recombinant human erythropoietin (rhEpo) and rhGM-CSF for the next 12 weeks. Five patients had refractory anemia (RA), 3 refractory anemia with ringed sideroblasts (RAS), and 5 refractory anemia with excess of blasts (RAEB). Ten patients were transfusion-dependent at the time of inclusion. Eleven patients completed this phase II study. Five responded with an increase in hemoglobin level (3 patients) or a reduction in transfusion requirement (2 patients). We registered no response in the remaining 6 patients during treatment. Patients responding to combined treatment had relatively low concentrations of plasma Epo and plasma ferritin before treatment with rhEpo and a normal karyotype throughout the study. Long-term bone marrow cultures did not predict the response. Still, responders seemed to have a higher number of colony-forming progenitors than nonresponders. In conclusion, combined therapy with rhGM-CSF and rhEpo may stimulate hematopoiesis and correct or improve anemia in some patients with MDS. PMID- 8237993 TI - Efficacy of intravenous immunoglobulin in the treatment of autoimmune hemolytic anemia: results in 73 patients. AB - To determine whether warm-antibody autoimmune hemolytic anemia (AIHA) responds to treatment with intravenous gammaglobulin (IVGG), we conducted separate pilot studies at three institutions enrolling a total of 37 patients. We combined these results with a review of 36 cases of AIHA treated with IVGG reported in the literature. Sixteen clinical variables were examined to determine associations with response to IVGG. Overall, 29 of 73 patients (39.7%) responded to IVGG therapy. Two variables were strongly related to a good response to IVIG: the presence of hepatomegaly (with and without splenomegaly) and a low pre-treatment hemoglobin. A trend towards a better response was observed in the 11 children. Overall, IVGG provided acute benefit in only 1/3 of patients and therefore cannot be recommended as standard therapy for AIHA. It may, however, be useful as adjunctive treatment in selected cases, such as in those with a pre-treatment hemoglobin < 6-7 gm/dl or those with hepatomegaly, and in clinical settings where the toxicity of other treatments may be an important consideration. PMID- 8237994 TI - Decreased fibrinolytic potential in patients with idiopathic avascular necrosis and transient osteoporosis of the hip. AB - A thrombotic etiology has been suggested as the cause of idiopathic avascular necrosis of the hip, although the underlying pathophysiological mechanisms are unknown. Transient osteoporosis of the hip has also been suggested to represent bone marrow edema that may be related to ischemia. We evaluated four patients with idiopathic avascular necrosis and one patient with transient osteoporosis of the hip for thrombotic potential placing a special emphasis on the fibrinolytic system. All five patients had identifiable abnormalities of fibrinolysis. Four patients had elevated levels of plasminogen activator inhibitor (PAI-1) and one patient had an inadequate increase in tissue plasminogen activator (tPA) post venous occlusion. Serum triglycerides were increased in three of the patients. These findings suggest an association between decreased fibrinolytic potential and the subsequent development of avascular necrosis and transient osteoporosis of the hip. These patients should have an evaluation of the fibrinolytic system with tPA and PAI-1 levels as well as a lipid profile. PMID- 8237995 TI - Regulation of megakaryocyte colony forming cell numbers and ploidy by dideoxynucleosides in immunodeficient mice. AB - We have recently demonstrated that azidothymidine (AZT) elevates the levels of circulating platelets in mice made immune deficient by infection with LP-BM5 murine leukemia virus (MAIDS mice). In an attempt to elucidate the mechanisms of the AZT platelet elevating effect, we examined the number of splenic and bone marrow megakaryocyte colony-forming cells (CFU-mk) and the ploidy of megakaryocytes derived from CFU-mk using fluorescence cytophotometric methods. Two other dideoxynucleosides (ddN) 2'3'-dideoxyinosine (ddl) and 2'3' dideoxycytidine (ddC) were assessed to determine the specificity of the effect of AZT. MAIDS mice were given ddN in drinking water for 15 days. AZT was the only ddN that significantly increased circulating platelet levels in MAIDS mice. AZT significantly increased splenic CFU-mk in MAIDS mice, but bone marrow CFU-mk were not affected. ddl and ddC failed to change either platelet levels or the numbers of splenic or bone marrow CFU-mk. The ploidy of megakaryocytes derived from splenic and bone marrow CFU-mk were examined by first identifying CFU-mk by staining for acetylcholinesterase, followed by nuclear staining with propidium iodide. The fluorescence of individual cells was then measured using a Perceptics image analysis system. Modal ploidy of CFU-mk megakaryocytes derived from spleen cells of AZT-treated immunodeficient mice was shifted upwards from 16N to 32N. Similarly, AZT treatment changed the modal ploidy number of colony megakaryocytes derived from bone marrows of immunodeficient mice from 16N to 32N. The ploidy distribution was also significantly shifted. ddl and ddC failed to significantly alter either modal ploidy number or distribution of megakaryocytes derived from splenic or bone marrow CFU-mk. These findings suggest that AZT may affect physiological processes that lead to platelet formation. PMID- 8237996 TI - Synergistic effect of storage pool deficient platelets and low plasma von Willebrand factor on the severity of the hemorrhagic diathesis in Hermansky Pudlak syndrome. AB - A study of 565 Puerto Rican patients with storage pool deficient (SPD) Hermansky Pudlak syndrome (HPS) demonstrated that most HPS patients had minor bleeding episodes while others had repeated, severe hemorrhagic episodes requiring transfusion. The severity of bleeding in these latter patients could not be attributed to their SPD alone. As swine with SPD platelets and low von Willebrand factor antigen (vWF:Ag) have more severe hemorrhages than pigs with either defect alone, 146 albino patients and 46 normally pigmented patients were examined for their level of vWF:Ag. The risk of SPD HPS patients having severe, repeated bleeding episodes increased when vWF:Ag fell below 70 U/dL. Family studies indicated that low vWF:Ag levels were more frequently associated with O blood group than from a gene suppressing production or release of vWF1. HPS patients should be tested for vWF:Ag levels. PMID- 8237997 TI - Hypofibrinolysis in patients with hypercoagulability: the roles of urokinase and of plasminogen activator inhibitor. AB - The prevalence of abnormalities of fibrinolysis in patients with venous thromboembolism is as yet unknown. Defined abnormalities include congenital dysfunction and deficiency of plasminogen, and probably impaired plasminogen activation secondary to elevated levels of plasminogen activator inhibitor type 1 (PAI-1) or to impaired release of tissue plasminogen activator (tPA). In this preliminary study, we analyzed plasma samples from 21 patients for whom an investigation for possible thrombophilia was requested. Twenty of the patients had venous thromboembolism, and one had arterial thrombosis at an early age. Two patients had deficiency of protein C or protein S, but no other recognized biochemical disturbances related to thrombophilia were identified. Patient samples and plasma from 25 normal controls were assayed for tPA activity, PAI-1 activity, and urokinase (uPA) activity and antigen. tPA activity and antigen were not significantly different in patients than in controls. PAI-1 activity was significantly greater in patients (P < 0.0001). uPA activity was not different in the two groups. However, uPA antigen was significantly reduced in patients compared to controls (P = 0.001). These data suggest that hypofibrinolysis leading to a risk of thrombosis may be caused not only by elevated PAI-1 activity but also by reduced total uPA concentration. PMID- 8237998 TI - Multimeric pattern of plasma and platelet von Willebrand factor is normal in uremic patients. AB - Several studies have reported increased von Willebrand factor (vWF) levels in plasma of uremic patients, with a normal multimeric pattern. Two recent studies, however, have shown a reduction of the larger multimers of plasma vWF and one has also found low levels of platelet vWF in uremic patients with prolonged bleeding time (BT). We have measured plasma and platelet vWF and analyzed its multimeric pattern in 20 uremic patients, 11 with a prolonged BT and 9 with a normal BT. For Plasma vWF, no difference for vWF:Ag, RiCof, and ristocetin-induced platelet agglutination was found between the two groups with normal and prolonged BT. The multimeric pattern of plasma vWF, as evaluated by densitometric scanning of the electrophoretic gels, was normal in both groups. For platelet vWF, vWF:Ag and RiCof content was similar in the two groups. The multimeric pattern was indistinguishable from that of normal platelets. In conclusion, our study does not confirm the presence of a structural defect of plasma vWF and the reduction of platelet vWF content in uremia. PMID- 8237999 TI - Hb Adana or alpha 2(59)(E8)Gly-->Asp beta 2, a severely unstable alpha 1-globin variant, observed in combination with the -(alpha)20.5 Kb alpha-thal-1 deletion in two Turkish patients. AB - We have identified a severely unstable hemoglobin variant through sequencing of amplified DNA involving the alpha 1-globin gene; the mutation is located in codon 59 (CCG CAG) and results in a Gly-->Asp replacement. This amino acid substitution concerns a glycine residue at an internal position in the E helix, which is in close contact with a glycine residue of the B helix; introduction of the larger and charged aspartic acid residue greatly affects the stability of the molecule. This variant was present in association with a common alpha-thalassemia-1 deletion [-(alpha)20.5 kb] in two adults and caused a severe type of Hb H disease with anemia, low levels of Hb A2, increased zeta chain, and Hb Bart's. In vitro chain synthesis in reticulocytes showed a high specific activity of the variant alpha chain. Only a minute quantity of Hb H was present but instead about 10% of Hb Bart's was observed. The increased synthesis of gamma chains was likely due to specific characteristics of a chromosome with haplotype #3, which was present in both patients. The same family was studied 18 years ago; the improved methodology presently available has led to a corrected diagnosis for these patients. PMID- 8238000 TI - Hemostatic molecular markers in nephrotic syndrome. AB - Quantitative changes of hemostatic molecular markers were studied in patients with nephrotic syndrome. The plasma levels of fibrinopeptide A (FPA), thrombin antithrombin III complex (TAT), products of thrombin activation, and fragment F1 + 2 (F1 + 2), a product of prothrombin activation, were measured by enzyme immunoassay in 21 patients with nephrotic syndrome and in 16 normal controls. The mean value of FPA was 17.5 +/- 7.5 ng/ml (mean +/- SD) in nephrotic patients and 4.5 +/- 0.3 ng/ml in normal controls (P < 0.02); F1 + 2 concentration was 1.4 +/- 0.3 ng/ml in nephrotic patients and 0.5 +/- 0.1 ng/ml in normal controls (P < 0.001); TAT level was 1.0 +/- 0.3 microgram/l in nephrotic patients and 0.2 +/- 0.1 microgram/l in normal controls (P < 0.05). These data indicated intravascular hemostasis activation. Based on these results, we propose that low antithrombin III level in nephrotic patients may be due to both urinary loss and intravascular consumption. PMID- 8238001 TI - Hemolytic anemia associated with lead poisoning from shotgun pellets and the response to Succimer treatment. AB - Symptomatic lead poisoning with severe hemolytic anemia was observed in a patient with retained shot gun pellets. Surgical resection of the retained pellets and the use of a newer chelating agent, Succimer (2,3-dimercaptosuccinic acid) successfully lowered blood lead level. Hemolytic anemia was associated with deficient erythrocyte pyrimidine 5'-nucleotidase, and lowering of the lead level corrected the deficiency, suggesting that the enzyme deficiency is responsible for the hemolysis associated with lead poisoning. This case illustrates that retained lead pellets from shotgun wounds can cause severe lead poisoning. PMID- 8238002 TI - Refractory thrombocytopenia in a 27-year-old female following allogeneic bone marrow transplantation. PMID- 8238003 TI - von Willebrand's disease and arteriosclerosis. PMID- 8238004 TI - Bone marrow aplasia associated with ticlopidine therapy. PMID- 8238005 TI - Lymphoma transformation in polycythaemia vera treated with hydroxyurea. PMID- 8238006 TI - Thrombocytopenia associated with use of Lorenzo's oil. PMID- 8238007 TI - Alport syndrome: from bedside to genome to bedside. AB - Alport syndrome is a genetic disorder of basement membranes manifested clinically by a progressive nephropathy and, in many families, sensorineural hearing loss and ocular lesions. During the 1980s evidence was amassed indicating type IV (basement membrane) collagen as the defective protein in Alport This hypothesis was confirmed in 1990 by the cloning of the X-chromosomal gene COL4A5, which encodes the alpha 5 chain of type IV collagen, and the discovery of mutations in this gene in many Alport kindreds. The results of results of recent studies suggest that the alpha 5(IV) chain forms a distinct collagenous network with the alpha 3 and alpha 4 chains of type IV collagen and that mutations in alpha 5(IV) may prevent the normal incorporation of alpha 3(IV) and alpha 4(IV) into basement membranes. Renal biopsy remains an important modality for making the diagnosis of Alport syndrome, but may eventually be replaced by molecular genetic techniques. Posttransplant anti-glomerular basement membrane nephritis occurs rarely in Alport patients and may be restricted to a subgroup with particular COL4A5 mutations. It is not clear why COL4A5 mutations result in glomerulosclerosis and renal failure, or whether this process may be slowed through dietary or pharmacologic intervention. PMID- 8238008 TI - Alport-leiomyomatosis syndrome: an update. AB - Alport-leiomyomatosis syndrome is a polygenic syndrome with a dominant X-linked inheritance pattern resulting from a large deletion in the 5' end of the COL4A5 gene coding for the type IV collagen alpha 5 chains. Hypothetically, the deletion extends beyond the 5' end and probably includes a second contiguous gene responsible for leiomyomatosis (the DL gene) and even a third one coding for congenital cataract (the CCT gene). PMID- 8238009 TI - Altered urinary excretion of lysosomal hydrolases in pregnancy. AB - Creatinine concentrations and the activities of five lysosomal hydrolases were measured in the serum and urine of 14 healthy nonpregnant control women and 19 healthy pregnant women. Fractional enzyme excretion (FEE) values for beta glucuronidase, beta-hexosaminidase, alpha-galactosidase, beta-galactosidase, and alpha-mannosidase were calculated and compared between the two groups of subjects. Fractional enzyme excretion was calculated as the ratio of enzyme clearance to creatinine clearance. The FEE values for beta-galactosidase and alpha-mannosidase between the nonpregnant and pregnant populations were not statistically different; however, relative to the nonpregnant control group, the median FEE values for beta-glucuronidase (P < 0.03), beta-hexosaminidase (P < 0.06), and alpha-galactosidase (P < 0.02) were decreased approximately 1.5-, 1.8 , and 2.7-fold, respectively, in the pregnant population. The median urinary beta galactosidase activity for the pregnant population, when expressed on the basis of creatinine, was twofold higher than that of the control group (P < 0.0005). These data indicate that with pregnancy there are marked changes in the urinary excretion of selected lysosomal enzymes, particularly alpha-galactosidase and beta-glucuronidase. When the molecular weights of these five hydrolases were compared between kidney homogenate and control urine, a correlation of 0.96 was observed, while the correlation between control serum and control urine was 0.69. This suggests that the FEE value differences between the pregnant and control groups are most likely due to changes in tubule cell metabolism, either decreased secretion or increased reabsorption. These biochemical changes may provide a means of assessing changes in renal function during pregnancy. PMID- 8238010 TI - Urinary epidermal growth factor levels in patients with acute renal failure. AB - To investigate the importance of epidermal growth factor (EGF) in patients with renal dysfunction, urinary human EGF (hEGF) levels were determined by radioimmunoassay in 16 patients with acute renal failure (ARF) and in 12 healthy controls. Seven patients with chronic renal failure also were studied. Urinary hEGF levels, corrected for urine creatinine concentrations, were significantly decreased in patients with ARF in the acute phase compared with normal control subjects (0.98 +/- 0.20 v 13.74 +/- 1.18 ng/mg creatinine, P < 0.001), and subsequently increased during the recovery phase (6.10 +/- 0.73 ng/mg creatinine, P < 0.001 v acute phase). A significant positive correlation existed between urinary hEGF levels and creatinine clearance in patients with ARF (r = 0.66, P < 0.001). Serum hEGF levels also were significantly lower in patients with ARF compared with normal control subjects (0.10 +/- 0.01 v 0.30 +/- 0.03 ng/mL, P < 0.001). No significant correlation was found between hEGF concentrations in serum and urine. In conclusion, measurement of urinary hEGF may be useful in the diagnosis of ARF and for following the recovery of the kidney after severe tubular injury. PMID- 8238011 TI - Reversible acute renal failure as an atypical presentation of ischemic nephropathy. AB - Ischemic nephropathy (IN) is defined as a clinically significant reduction in glomerular filtration rate in patients with hemodynamically significant obstruction to renal artery flow in a solitary functioning kidney or with bilateral renal artery stenosis (RAS). Ischemic nephropathy typically has a subacute to chronic course that may lead to end-stage renal disease. Acute anuric renal failure, which occurs less commonly with IN, is usually associated with moderately severe hypertension and has been attributed to certain risk factors: angiotensin-converting enzyme inhibition, a reduction in blood pressure secondary to antihypertensives or volume contraction, and exposure to contrast media. We present a series of six patients with IN and acute, anuric renal failure without either moderately severe hypertension or the previously defined risk factors. Of these six patients, four had RAS in a solitary kidney and two were found by ultrasound to have disparity in kidney size and bilateral RAS. Within 1 week of surgery, three patients developed renal failure that did not involve the kidney(s) responsible for the anuria and thus mimicked postoperative acute renal failure. Creatinine levels pre-anuria (1.2 to 2.1 mg/dL), during renal failure (5.0 to 12.8 mg/dL), and postrecovery (1.6 to 2.8 mg/dL) showed recovery of renal function, with renal artery bypass in four patients (sustained at 1 year). Two patients refused surgery and are on chronic dialysis. Acute renal failure in IN may occur postoperatively or spontaneously, and emergent intervention (ultrasound, angiography, angioplasty, and/or surgery) in this setting may lead to the correction of RAS and preservation of renal function. PMID- 8238012 TI - Reversible activation defect of the platelet glycoprotein IIb-IIIa complex in patients with uremia. AB - Patients with chronic renal failure may experience a bleeding tendency and blood loss after surgical procedures or trauma. Altered platelet function has been indicated as the major cause of uremic bleeding, but its pathogenesis remains to be clarified. In two groups of patients with chronic renal disease of various etiology, the receptor function of glycoprotein (GP) Ib and GP IIb-IIIa complex was studied. Glycoprotein Ib was assessed with both 125I-von Willebrand factor (vWF) and 125I-asialo-vWF binding to platelets. Activation-dependent receptor function of the GP IIb-IIIa complex was studied with 125I-fibrinogen and 125I-vWF binding to washed platelets stimulated with adenosine diphosphate plus epinephrine (10 mumol/L each). Flow cytometric analyses on resting and stimulated platelets were performed using an activation-dependent, anti-GP IIb-IIIa monoclonal antibody (PAC1) as well as an activation-independent antibody (LJ-P1). Binding of PAC1 also was assessed in washed and stimulated platelets and in platelet-rich plasma before and after dialysis. We found that the activation dependent receptor function of the GP IIb-IIIa complex is defective in uremia, as shown by decreased binding of both vWF and fibrinogen to stimulated platelets. Moreover, binding of the activation-dependent anti-GP IIb-IIIa monoclonal antibody, PAC1, was significantly decreased in uremia compared with that of the activation-independent antibody, LJ-P1. Thus, the number of GP IIb-IIIa receptors expressed on the platelet membrane is normal, but their activation is impaired. In contrast to the functional abnormality of GP IIb-IIIa, the vWF-binding activity of GP Ib was normal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238013 TI - Morphologic features of the myopathy associated with chronic renal failure. AB - Our previous studies indicate that impaired function of skeletal muscle limits the exercise tolerance of patients with end-stage renal failure who are either maintained on dialysis or undergo renal transplantation. To study the morphology of the condition, muscle biopsies were performed on eight patients with renal failure-associated myopathy. Control samples were taken from seven healthy athletes undergoing knee surgery and from five otherwise healthy but untrained subjects. Tissues were examined by routine light and transmission electron microscopy. Histochemical staining of frozen sections for myosin adenosine triphosphatase and quantitative computer-assisted morphometry of the fiber type and size was performed. The mean (+/- SD) size for type I fibers in patients was 61.2 +/- 11.8 microns, while type II fibers measured 46.7 +/- 11.4 microns. The mean percentage of type II fibers was 67% +/- 12%. These values are within the normal population range and were not different from controls. Significant changes were found on light microscopy of patient samples. These included fiber splitting, internalized nuclei, nuclear knots, moth-eaten fibers, fiber degeneration and regeneration, increased content of lipid droplets, and fiber type grouping. Electron microscopy showed a large variety of nonspecific abnormalities, including mitochondrial changes, Z-band degeneration, myofilament loss, and accumulation of intracellular glycogen. Ten of 12 control subjects showed no such changes; minor abnormalities were noted on both light and electron microscopy in the remaining two subjects. Muscle oxidative capacity (19.5 +/- 5.1 microL O2/min) for patients with end-stage renal failure was not different from values for those who had undergone renal transplantation, but was lower than values found in trained athletes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238014 TI - The effect of treatment with recombinant human erythropoietin on skeletal muscle function in patients with end-stage renal failure treated with regular hospital hemodialysis. AB - The effect of treatment with recombinant human erythropoietin on skeletal muscle was investigated in 11 anemic transfusion-dependent patients on the hospital hemodialysis program. There were no significant changes in anthropomorphic measurements during the study, but the maximum voluntary contraction increased significantly for each muscle group studied (pre-erythropoietin and post erythropoietin values, respectively, were as follows: biceps, median 170 N [range, 83 to 220 N] v 189 N [range, 89 to 245 N]; triceps, 88 N [range, 59 to 167 N] v 106 N [range, 95 to 185 N]; deltoid, 168 N [range, 78 to 247 N] v 193 N [range, 93 to 290 N]; and quadriceps, 202 N [range, 165 to 300 N] v 265 N [range, 185 to 335 N]; P < 0.05). There were no significant changes in muscle strength in a control group of regular hemodialysis patients. Programmed electrical stimulation of the quadriceps following erythropoietin treatment resulted in both a greater force generated and a longer duration of contraction. Following cessation of the electrical stimulus the relaxation rate for the type II fibers was quicker following erythropoietin therapy. This suggests that some of the benefit observed in physical well-being following correction of the anemia of chronic renal failure with erythropoietin is due to an improvement in voluntary skeletal muscle function. PMID- 8238015 TI - Synthesis of beta 2-microglobulin in lymphocyte culture: role of hemodialysis, dialysis membranes, dialysis-amyloidosis, and lymphokines. AB - Dialysis-amyloidosis (A beta 2M) is a recently recognized chronic complication in long-term dialysis patients, apparently effecting 5% to 10% of all dialysis patients. In 1985, Gejyo et al (Biochem Biophys Res Commun 129:701-706, 1085) and Shirahama et al (Lab Invest 53:705-709, 1985) identified beta 2-microglobulin (beta 2-M) as the major constituent protein of this unique type of systemic amyloidosis. The specific pathogenesis of A beta 2M remains unknown, although beta 2-M has been clearly identified as playing a central role as the amyloidogenic protein. To investigate the factors responsible for in vitro beta 2 M synthesis, we studied beta 2-M production by lymphocyte cultures obtained from dialysis patients and grown under a variety of different conditions, and compared the results to a control group of subjects with normal renal function. We could not demonstrate any stimulatory influence on beta 2-M synthesis by the hemodialysis treatment, the type of dialysis membrane used, or the clinical presence of A beta 2M. Rather, dialysis membranes, sterilized with ethylene oxide or gamma rays, added to the lymphocyte cultures exerted a strong dose-dependent inhibitory effect on beta 2-M synthesis. From the results of this study, we conclude that peripheral blood lymphocytes in uremic patients synthesize beta 2-M normally and that the direct interaction between circulating lymphocytes and the dialysis membrane that occurs during hemodialysis does not seem to contribute directly to beta 2-M synthesis. PMID- 8238016 TI - Tidal peritoneal dialysis: kinetics and protein balance. AB - Some patients find automated peritoneal dialysis preferable to continuous ambulatory peritoneal dialysis (CAPD). Unfortunately, automated peritoneal dialysis prescriptions are time consuming and can impede rehabilitation. We wished to determine whether an 8-hour tidal peritoneal dialysis (TPD) prescription could maintain the time averaged blood urea nitrogen at 60 mg/dL or less while patients consumed a diet containing approximately 1.2 g protein/kg body weight/d. Ten home dialysis patients previously stabilized on continuous cyclic peritoneal dialysis volunteered for a metabolic balance study conducted at the University of Iowa's Clinical Research Center. A peritoneal equilibration test was conducted and mass transfer area coefficients (MTaCs) were derived for each subject. Nitrogen balance was measured during the last 5 days of a 12-day constant diet while patients underwent a series of monitored nocturnal dialyses. Mass transfer area coefficient measurements were reproducible and independent of the filling volume and ultrafiltration, but varied between subjects (normalized MTaCurea = 33.6 +/- 16.3 mL/min, normalized MTaCcrt = 16.3 +/- 9.5 mL/min). Tidal peritoneal dialysis urea and creatinine clearances could be predicted by these MTaC values (r2 = 0.70 urea, r2 = 0.91 creatinine). Nitrogen balance assumptions predicted, and we confirmed, a relationship between dietary protein intake and urea nitrogen generation (r2 = 0.82) during TPD. A normalized protein catabolic rate of 1.2 g/kg/d resulted in a urea nitrogen generation rate of approximately 100 mg/kg/d. If a patient's protein intake was approximately 1.2 g/kg/d, then TPD with a weekly urea clearance normalized to body volume (Kt/V(urea)) of approximately 2.1 (urea clearance, approximately 0.35 mL/kg/min) could maintain a time averaged blood urea nitrogen of approximately 60 mg/dL.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238017 TI - Treatment of Staphylococcus aureus nasal carriers in continuous ambulatory peritoneal dialysis with mupirocin: long-term results. AB - We present the clinical results of a prospective protocol of the treatment of Staphylococcus aureus nasal carriers (SANCs) in our continuous ambulatory peritoneal dialysis unit with mupirocin (Bactroban, SmithKline Beecham Pharmaceuticals, Philadelphia, PA). We monitored the incidence of peritonitis and catheter exit-site infection, the rate of infection-related catheter loss, and the degree of association between SANC state and S aureus infection. The study group included 94 patients with a follow-up of 1,097 patient-months (phase B). The same information was retrospectively collected among 74 continuous ambulatory peritoneal dialysis patients treated during the 24 months preceding the study period (follow-up of 1,043 patient-months) (phase A). S aureus nasal carriage was observed in 47.5% of the patients. Mupirocin was very effective in eradicating S aureus from the nares, but most patients required periodic retreatment. The incidence of S aureus peritonitis decreased from 1 episode/58 patient-months in phase A to 1 episode/548 patient-months in phase B, and the incidence of exit site infection decreased from one episode/55 patient-months in phase A to 1 episode/137 patient-months in phase B. However, there was a simultaneous increase in the incidence of infections by other gram-positive and -negative bacteria. The rate of catheter loss after peritonitis (P = not significant) or exit-site infection (P < 0.05) tended to decrease from phase A to phase B. Seventy-seven percent of the peritonitis infections and 74% of the exit-site infections by S aureus occurred in SANCs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238018 TI - Pulmonary-renal syndrome in a patient with fibrillary glomerulonephritis. AB - A patient presented with a rapid deterioration in renal function requiring dialysis. Renal biopsy demonstrated fibrillary glomerulonephritis. The patient was treated with steroids and cyclophosphamide and recovered sufficient renal function to discontinue dialysis. However, as immunosuppressive therapy was withdrawn, renal function worsened and the patient developed pulmonary hemorrhage and respiratory distress. Immunosuppressive therapy was reinstituted. During treatment the patient's renal function improved and pulmonary manifestations resolved. This case is further evidence that fibrillary glomerulonephritis can present as a pulmonary-renal syndrome and can respond to aggressive treatment with immunosuppressive agents. PMID- 8238019 TI - Paraparesis due to massive ectopic paravertebral calcification in a patient on maintenance hemodialysis. AB - Secondary hyperparathyroidism is ubiquitous in patients with end-stage renal disease who are on dialysis. We report a dialysis patient with an unusual manifestation of hyperparathyroidism. Initially, the patient was misdiagnosed to have clavicular osteomyelitis. The underlying hyperparathyroidism was not recognized, only to be discovered later, when the patient presented with spastic paraplegia and massive ectopic calcification in the cervical spine. The patient made a complete recovery following surgical resection and parathyroidectomy. PMID- 8238020 TI - The role of insulin-like growth factors in diabetic kidney disease. AB - Early renal manifestations of type I diabetes include kidney enlargement, increased glomerular filtration rate, and renal plasma flow. These hemodynamic changes may be caused by a number of factors, including growth hormone and/or insulin-like growth factor-I (IGF-I). Streptozotocin-induced insulinopenic diabetes in rats represents a model of human type I diabetes and is associated with the early hemodynamic changes in the kidney seen in poorly controlled type I diabetic patients. These changes are preceded by an accumulation of IGF-I peptide in the kidney. Insulin-like growth factor-I is not locally produced, but rather accumulates from circulating IGF-I, trapped by increased levels of IGF-binding proteins, particularly IGF-binding protein-1. The hemodynamic effects, reproduced by infusions of recombinant human IGF-I in normal rats, may be blocked by co infusion of a kinin-receptor antagonist, suggesting that at least one of the mechanisms involved is the kallikrein-kinin system. These studies strongly support the notion that the IGF system may play a role in early hemodynamic manifestations of the diabetic kidney. Whether these effects lead to long-term diabetic renal disease remains to be studied. PMID- 8238021 TI - Eicosanoids in the pathogenesis of the functional and structural alterations of the kidney in diabetes. AB - Diabetes mellitus alters the cellular production of eicosanoids in a number of tissues, including the kidney, and these agents have in turn been implicated in the pathogenesis of diabetic nephropathy. As delineated in the streptozotocin diabetic rat (SDR) model, a preferential enhancement of glomerular synthesis of the vasodilatory prostaglandins (PGs) PGE2 and PGI2 with concurrent smaller increases in thromboxane (TX)A2 occurs within 1 week after induction of diabetes. This early alteration in glomerular synthesis of eicosanoids in the SDR has been linked to glucose-induced activation of the glomerular protein kinase C signalling system that enhances phospholipase A2 activity and, therefore, release of membrane-bound arachidonic acid for oxygenation. The preferential increase in glomerular production of vasodilatory PGs may contribute to the glomerular hyperfiltration that is characteristic of early diabetes. After more prolonged (months) diabetes in the SDR, glomerular generation and urinary excretion of thromboxane (TX) are preferentially enhanced. Studies with selective inhibitors of TX synthesis in the SDR have implicated this eicosanoid in the pathogenesis of both albuminuria and glomerular structural changes (basement membrane thickening and mesangial matrix expansion). Direct stimulation of matrix protein production has been demonstrated in cultured mesangial cells in response to both TX and high ambient concentrations of glucose. The actions of TX and glucose on mesangial cell matrix production appear to be interactive, with each signalled through distinct pathways of protein kinase C activation. PMID- 8238022 TI - The extracellular matrix in diabetic nephropathy. AB - In the subgroup of diabetic patients who are destined to develop the full spectrum of the clinical syndrome of diabetic nephropathy, the kidney is afflicted with a series of distinct structural lesions principally involving the extracellular matrices. Diabetic nephropathy is characterized by hypertrophy of both glomerular and tubular elements, progressive accumulation of extracellular matrix components in the glomerular mesangium, and thickening of the glomerular and tubular basement membranes. Albeit less well recognized, progressive tubulointerstitial fibrosis is also a feature of the syndrome. Irrespective of pathogenetic mechanisms (be they metabolic, hemodynamic, or genetic), the structural changes involving the renal extracellular matrix compartments are believed to be the basis for the appearance of overt dysfunction, namely, proteinuria, hypertension, and renal failure. Therefore, a full understanding of the mechanisms that culminate in irreversible kidney failure requires a closer inspection of the status of the extracellular matrix in diabetes. This review outlines the different structural changes that typically occur during the course of the disease. Both glomerular and tubulointerstitial changes are reviewed. Valuable structural-functional correlations have been derived from examining kidney specimens obtained from patients with a wide spectrum of disease stages. Experimental animal models, supplanted with recent investigations in tissue culture on the effect of high ambient glucose levels, have increased our understanding of the cellular mechanisms that underlie the disordered matrix composition. Alterations in the metabolism of the collagens, proteoglycans, and other matrix constituents are reviewed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238023 TI - The biology of angiotensin II receptors. AB - Angiotensin II is very important in the regulation of blood pressure. This small peptide binds to cell surface receptors, initiating a wide diversity of physiologic responses. There are two major subtypes of angiotensin II receptors referred to as AT1 and AT2. In this article we describe the cloning and the biochemical characterization of the AT1 receptor. Antibodies against this receptor have been used to define its tissue distribution. The AT1 receptor is a member of the seven transmembrane spanning class of receptors. It initiates a complex series of signaling events, including activation of membrane phospholipases and intracellular kinases. In the human a single AT1 receptor protein mediates virtually all the effects of angiotensin II, suggesting that tissue specificity of angiotensin II must be due to organ-specific intracellular signaling. PMID- 8238024 TI - Pulmonary edema and left ventricular hypertrophy. PMID- 8238025 TI - Child labor: a re-emergent threat. PMID- 8238026 TI - Regulation of child labor revisited. PMID- 8238027 TI - Youth, race, and labor: working kids and historical ambivalence in twentieth century America. AB - As a society, we share assumptions that exhibit a profound historical ambivalence regarding young people at work. On the one hand, we all acknowledge that there is something socially and morally reprehensible about forcing children to toil. Many of us see child labor as a vestigial remnant of the harshest aspects of the Industrial Revolution. On the other hand, we also continue to see work as a sign of the moral and social health of a community and hence, we see employment among the young as a stabilizing and redemptive activity. Especially when addressing the nation's poor, urban and African-American populations, youth unemployment is almost universally understood as a root cause of social unrest and community breakdown. This paper outlines some of the historical arguments about children in the work force and raises the question about the national resolve to seriously address this issue. PMID- 8238028 TI - Health hazards to children in agriculture. AB - Children comprise a significant portion of the agricultural workforce and are exposed to many workplace hazards, including farm machinery, pesticides, poor field sanitation, unsafe transportation, and fatigue from doing physically demanding work for long periods. Migrant farmworker children face the additional hazard of substandard or nonexistent housing in the fields. Children account for a disproportionate share of agricultural workplace fatalities and disabling injuries, with more than 300 deaths and 27,000 injuries per year. The most common cause of fatal and nonfatal injury among children in agriculture is farm machinery, with tractors accounting for the greatest number. Remedies to the problems of child labor must take into account family economics and the need for child care. Labor law reform and rigorous enforcement of existing laws and of workplace health and safety requirements are vital to better protect the children and adults working in agriculture. PMID- 8238029 TI - Health hazards to children in the service industries. AB - The purpose of this article is to examine the persistent, yet underrecognized, problem of adolescent occupational injury. There is little appreciation for the magnitude and scope of this problem, including, particularly, the underlying causes and factors of adolescent occupational injury; this is especially so outside the occupational health field. There are many reasons why the scope of adolescent occupational injury remains unappreciated. For example, injury surveillance systems, including workers' compensation records, do not reveal the level of this problem because many injuries are not properly compensated. This paper examines health hazards to children in the service industries from various perspectives. This scrutiny will include an examination of adolescent occupational injury levels and the reasons why this problem is large and growing. In closing, we consider ways to improve public policies in order to portray this problem accurately, the first step toward meaningful injury prevention strategies. PMID- 8238030 TI - Child and adolescent injury in the United States: how occupational injuries fit in. AB - Injury is the leading cause of death in adolescence, with the highest rates for 15- to 19-year-olds, an age span also marked, for many, by entry into the workforce. Occupational injuries affecting youth occur in the context of the larger problem of injury affecting children and adolescents. An understanding of the broader context can help to guide efforts to control occupational injuries. Nonfatal injuries vastly exceed fatal ones, but more is known about the latter. Injuries are the leading cause of death in the U.S. for ages 1 to 44 years. The proportion of child and adolescent deaths due to injury has risen for several decades; injury rates have been stable while deaths from natural causes have declined. Injury patterns vary with age, race, and sex, and also internationally. The two leading causes of injury deaths for U.S. adolescents are motor vehicle occupant injury (related largely to alcohol) and homicide (related largely to firearms). Contemporary injury prevention approaches focus on means to prevent the transfer of damaging amounts of energy to potential victims. The most effective approaches are passive (engineering/environmental) and/or require one time-only behaviors. Prevention of child and adolescent occupational injuries will need to build on approaches that have been successful for other child and adolescent injuries, as well ones that have been successful for adult occupational injuries. A multidisciplinary approach will be needed. PMID- 8238031 TI - Work-related injuries among Massachusetts children: a study based on emergency department data. AB - Millions of children in the United States work and, despite federal and state laws, face safety hazards. Previous studies have documented large numbers of injuries suffered on the job by working children. This study describes work related injuries to children 14-17 years old in 14 Massachusetts communities (representing 5% of the state population) based on data from emergency departments and hospital admissions collected as part of a large population-based surveillance study of injuries to children and adolescents from 1979 to 1982. An estimated 1,176 work-related injuries occurred during the three-year period, accounting for 7-13% of all injury-related emergency department visits in this age group; the proportion among 17-year-olds was 14-26%. Cuts and lacerations were the most frequent type of injury, cutting/piercing was the most common cause, and cutting instruments were the most frequently identified products. Seventeen children were hospitalized for work-related injuries during the course of the study. The estimated annual rate of occupational injury rose from 3.7/1,000 children for 14- to 15-year-olds to 44.7/1000 for 17-year-olds; these rates count all resident children, regardless of their employment status, in the denominator. Rates based on actual hours worked are much higher, and strongly suggest that working minors should be considered a high-risk group for occupational injury. This study adds to the mounting evidence that work-related injuries to children are a significant public health problem and contribute significantly to the overall incidence of injuries among children 14-17 years old. The study also demonstrates the potential of emergency department data as a valuable source of information about work-related injuries to children. Active surveillance systems combining data from emergency departments, workers' compensation, and other potential sources should be established to fully document the nature and extent of the problem. Efforts to prevent these injuries will require the combined attention of employers, parents, medical providers, educators and regulators, as well as working children themselves. PMID- 8238032 TI - Children at work: prevention of occupational injury and disease. AB - Because children are an isolated population that generally lacks a collective political voice, it is up to the rest of society to look after their well-being. The grim economic circumstances that plague impoverished nations around the world have resulted in many young children having to work to help their families survive. Often, these children have no choice but to work in dangerous places and under generally appalling conditions. Even in wealthy countries like the United States, the problems associated with child labor are a legitimate threat to our single most important investment for the future--the safety and health of our children. PMID- 8238033 TI - Remedies to the problem of child labor: the situation in the apparel industry. AB - When you realize how long the problem of child labor has been around, anyone who ventures into the terrain of remedies obviously needs a long memory and not a little optimism. What have we tried? What has worked? And what has not worked? To answer these questions, we must first look at how we have diagnosed the problem. Some say that the return of child labor is due to the present recession. Hard pressed businesses are looking for cheap and cheaper labor. Sweatshops proliferate. When the recession recedes, so will child labor. If it were that simple, we could all congratulate ourselves on having conducted this enlightened symposium and go home without worrying much more about the problem. The magic hand of the market, in due course, will straighten it all out. Let me tell you something about the apparel industry in New York where new laws and strict enforcement make the only difference. Over 80% of OSHA inspections were triggered by the state's Apparel Task Force. PMID- 8238034 TI - Child labor legislation in New York State: a 1990s update. AB - Presented here are the salient features of the provisions of the child labor law that New York State passed. The article confronts a basic question: "Should we have child labor at all?" PMID- 8238035 TI - Congressional approaches toward remedies to problems of child labor. AB - Hearings by the Employment and Housing Subcommittee and nationwide "sweeps" by the Labor Department focused attention in 1990 on the growing problem of child labor. Fast food and retail establishments employ the majority of minors and were found to have the most violations. Although the bulk of violations affected hours of work (too many or too late), many involved prohibited hazardous occupations. Reform measures were introduced in the House and the Senate. They would limit hours of work during school weeks for 16-17-year-olds, require school work permits, improve protection of farm workers, increase civil and criminal penalties for willful violations, and provide for national reports on employment of and injuries to workers under age 18. As of May 1992, the Senate Committee on Labor and Human Resources has voted out its bill, but floor action has not been scheduled. The House Subcommittee on Employment Standards has taken no action. PMID- 8238036 TI - Current trends in state child labor legislation and enforcement. AB - There is much current child labor law activity, none of it reflecting concern with occupational health hazards. The focus of the legislation is on time for and dedication to education as youth's foremost priority. The only areas of state legislative change that could affect children's health and safety in the workplace are a general movement toward monetarily increasing fines and penalties and some minor inclusions of agricultural restrictions on young workers. Enforcement efforts are in all cases a primary indication of state commitment to child labor law protections, and these efforts are decreasing. PMID- 8238037 TI - Substance abuse exacting toll on Medicaid. PMID- 8238038 TI - Heterosexual contact displaces drug abuse as main source of AIDS in women. PMID- 8238039 TI - Young women are more susceptible to AIDS, U.N. study reveals. PMID- 8238040 TI - Preparing an employee for retirement. PMID- 8238041 TI - Dealing with efficiency experts. PMID- 8238042 TI - Incompatibility of cefotaxime sodium and vancomycin sulfate during Y-site administration. PMID- 8238043 TI - Obtaining reimbursement for oncology pharmacy services provided to ambulatory patients. PMID- 8238044 TI - First dose at home. PMID- 8238045 TI - In-flight administration of methylprednisolone to patients with spinal cord injury. PMID- 8238046 TI - Cost avoidance, financial impact, and patient care. PMID- 8238047 TI - Ethics of maintaining patient confidentiality. PMID- 8238048 TI - Heroin and cancer pain initiatives. PMID- 8238049 TI - Spurious authorship. PMID- 8238050 TI - Personal computer-based expert system for quality assurance of antimicrobial therapy. AB - A personal computer (PC)-based expert system developed to monitor the appropriateness of antimicrobial therapy is described. Susceptibility test data and antimicrobial therapy data are downloaded daily from the microbiology department and pharmacy department computer systems. Relational database software allows for the indexing, sorting, and manipulation necessary for analysis. The expert system accomplishes its analyses using (1) databases of organisms, antimicrobial drugs, and susceptibility cutoff values, (2) programs for evaluating pathogenicity and therapy, and (3) algorithms that determine the timing and sequence of the analysis. System output consists of discrepant therapy reports that indicate that no therapy is being given despite the presence of pathogens, that the pathogens isolated are resistant to the therapy being given, that the therapy cannot be matched with susceptibility data on the isolates, or that the therapy was discontinued too quickly. The expert system has been in continuous operation at an 800-bed referral hospital since July 1991. During an 11-month period, the system generated 1538 discrepant therapy reports. The percentage of isolates resulting in reports varied substantially with the source of the isolate. Therapy was more likely to be improved when the physician was contacted about the potential problem indicated by the report than when the physician was not contacted. A PC-based expert system using data from unlinked pharmacy and microbiology computer systems automatically evaluated the appropriateness of antimicrobial drug therapy in light of susceptibility test data. PMID- 8238051 TI - Pharmaceutical services in the treatment of hemophiliac patients. AB - Pharmaceutical services in the treatment of hemophiliac patients in Sweden are described. To prevent arthropathy and improve quality of life, patients with severe hemophilia receive prophylactic treatment. Hemophiliac patients are referred for diagnosis, treatment, and follow-up to one of three hemophilia treatment centers. Patients are seen once or twice yearly for routine checkups. A new system for the distribution of antihemophilic factor and factor IX has been implemented to improve availability and reduce risks and costs. Patients registered at a center receive antihemophilic factor or factor IX from the corresponding hospital pharmacy. The number of different manufactured batches to which a patient can be exposed is restricted, and complete distribution data are recorded for each patient, enabling follow-up studies and facilitating withdrawals of faulty batches. Distribution of emergency kits to patients who do not receive supervised home therapy ensures the availability of treatment for bleeding episodes. Two studies performed to explore ways to reduce drug waste led to improvements in patient information, product guidelines, and equipment and a better understanding of the problems associated with product changes. Specialized pharmaceutical services in Sweden contributed to successful hemophilia treatment and reduced its cost. PMID- 8238052 TI - Use of i.v. cimetidine, ranitidine, and famotidine in 40 hospitals. AB - The drug experiences of 1200 patients receiving i.v. histamine H2-receptor antagonists were studied. Forty hospitals in five southeastern states participated in a drug-use evaluation (DUE). In addition to supplying background information, a pharmacist at each hospital used a standardized form to perform a concurrent review of 30 consecutive adult patients who had been started on i.v. cimetidine, ranitidine, or famotidine. In addition to the patient's age and the prescriber's medical specialty, specific DUE criteria included the reason given in the medical record for use; dosage regimen and adjustments made on the basis of the patient's renal function; other GI drugs taken concurrently; pharmacist intervention; simultaneous use of oral medications; occurrence of adverse events; H2-antagonist use with specified drug products known to affect serum drug concentrations of one or both medications; and use of gastric pH monitoring. Therapy with i.v. H2 antagonists was usually started by internal medicine specialists or surgeons, and most of the evaluated patients received such therapy for prevention of stress-related mucosal damage. According to estimated creatinine clearance, 34% of patients were in need of dosage adjustments, but such adjustments were made in only 49% of these. Forty percent of patients began therapy while taking at least one medication orally, and almost one fourth of patients were receiving an additional drug for the treatment of an acid-peptic or related indication. The occurrence of adverse events was similar for the three H2 antagonists studied. The findings identify several problematic areas in the way H2 antagonists are used in clinical practice. PMID- 8238053 TI - Authorship on a manuscript intended for publication. PMID- 8238054 TI - Pharmacy and therapeutics subcommittee on pediatrics. PMID- 8238055 TI - Pharmacist-physician drug fair for educating physicians in cost-effective prescribing. PMID- 8238056 TI - Specific gravity-based determination of dextrose content of total parenteral nutrient solutions for neonates. PMID- 8238057 TI - Stability of ceftriaxone sodium in infusion-pump syringes. PMID- 8238058 TI - Absence of liability in Illinois for failing to warn of a drug interaction. PMID- 8238059 TI - Use of pharmaceutical care plans in home i.v. therapy. PMID- 8238060 TI - Dentistry: it is safe and medically necessary! PMID- 8238061 TI - Thoracic complications of dental surgical procedures: hazards of the dental drill. AB - CASE REPORTS: Dental surgical procedures occasionally result in intrathoracic complications that may subsequently be encountered by clinicians. We report four patients with such complications, including pneumomediastinum, fatal descending necrotizing mediastinitis, and Lemierre's syndrome. In each of these patients, the commonly used dental handpiece with exhausted air directed to the working drill point was an important, but unrecognized, predisposition to their intrathoracic complication. CONCLUSION: Clinicians should be aware of the spectrum of these problems and, in particular, of the potential hazards of pressurized nonsterile air blown into open surgical sites by the dental drill. PMID- 8238062 TI - CD8 alveolitis in sarcoidosis: incidence, phenotypic characteristics, and clinical features. AB - PURPOSE: Although the accumulation of CD4 cells in the lung and other involved tissues is regarded as the distinctive immunologic feature of sarcoidosis, a few sarcoid patients can present with CD8 alveolitis. This study evaluates the incidence as well as the clinical and immunologic features of sarcoidosis presenting with CD8 alveolitis. PATIENTS AND METHODS: A total of 2,214 consecutive bronchoalveolar lavage (BAL) specimens obtained from 481 patients with sarcoidosis between January 1985 and December 1991 were retrospectively analyzed. Subjects who entered the study had the following characteristics: (1) lymphocyte alveolitis and (2) lung CD4/CD8 ratio less than 1.0. Only data obtained from patients with a first episode of pulmonary involvement were included in the analysis (394 patients). RESULTS: Fifteen of the 394 patients studied at the time of diagnosis showed CD8 alveolitis as the presenting manifestation; the incidence of this phenomenon was 3.8%. A follow-up study of BAL T-cell subsets demonstrated that patients who showed high-intensity CD8 alveolitis at the onset of the disease maintained the CD8 pattern of alveolitis during relapses. Phenotypic analysis of lung T cells revealed that the accumulation of CD8 lymphocytes was due to the discrete local increase of CD45RO+ "memory" cells equipped with a number of accessory structures, including adhesion molecules and class II major histocompatibility complex-related HLA-DR antigen. CONCLUSIONS: The accumulation of CD8 cells in the sarcoid lung is likely to reflect a homing of memory cells due to the ongoing immunologic response against the unknown antigen causing the disease. Although CD8 alveolitis can be considered a relatively rare event in sarcoidosis, the possibility that an increase of CD8 cells in the BAL fluid might be sustained by an underlying sarcoid inflammatory process should never be dismissed on clinical grounds in patients with interstitial lung disease. PMID- 8238063 TI - The economic burden of unrecognized vasodepressor syncope. AB - BACKGROUND: The objective of this study was to describe the cost of prior diagnostic evaluation in patients referred for evaluation of syncope whose history was typical of vasodepressor syncope. METHODS AND RESULTS: Thirty consecutive patients who were referred for evaluation of syncope of undetermined origin and whose history was highly suggestive of vasodepressor syncope participated in this study. These 30 patients represented 19% of 158 patients referred for evaluation of syncope during the period of enrollment. All patients had positive results of an upright-tilt test, confirming the diagnosis of vasodepressor syncope. At the time of evaluation, the type and results of all diagnostic tests that had been performed prior to referral were recorded for each patient. The cost of diagnostic testing was then determined based on the 1991 cost of these tests at the University of Michigan Medical Center. A mean of 4 +/- 2 major diagnostic tests were performed before referral to the University of Michigan Medical Center. The mean and median costs of diagnostic testing per patient prior to referral were $3,763 +/- 3,820 and $2,678 (range: 0 to $16,606) respectively. Six patients underwent no major diagnostic tests prior to referral and, therefore, the cost of major diagnostic testing was zero in these patients. In the remaining patients, the mean and median costs of diagnostic testing per patient were $4,704 +/- 3,713 and $3,777 (range: $1,025 to $16,606) respectively. CONCLUSIONS: The results of this study demonstrate that a diagnosis of vasodepressor syncope can be established clinically in approximately 20% of patients referred to a university hospital for evaluation of syncope of undetermined origin. Failure to recognize the clinical features of vasodepressor syncope in these patients resulted in up to $16,000 of unnecessary diagnostic testing. A greater awareness of the clinical features of vasodepressor syncope may, therefore, result in significant economic savings. PMID- 8238064 TI - Relationship of apolipoprotein E phenotypes to hypocholesterolemia. AB - PURPOSE: Persons with total cholesterol (TC) levels less than 130 mg/dL (less than 3.26 mmol/L) make up less than 1% of a healthy population. Causes of hypocholesterolemia include a diet very low in cholesterol and saturated fat, disease, genetic factors (including low apolipoprotein B-100 [apo B-100] and the apo E allele), and drug therapy. The purpose of this study was to determine the causes of hypocholesterolemia in a healthy Kaiser Foundation Health Plan (KFHP) population. PATIENTS AND METHODS: We conducted a dietary and health survey of 201 healthy hypocholesterolemic adults (range: 2.04 to 3.88 mmol/L [79 to 150 mg/dL]) and 200 matched control subjects with TC levels in the middle quintile of the population (range: 5.0 to 5.61 mmol/L [194 to 217 mg/dL]) who had routine health screening from 1983 through 1985. We did apo E phenotyping studies and lipid and apo A-1 and B-100 measurements in a subgroup of 45 hypocholesterolemic subjects (mean TC level: 3.26 mmol/L [126 mg/dL]) and in a comparison group of 49 unmatched volunteers (mean TC level: 5.04 +/- 0.75 mmol/L [195 +/- 29 mg/dL]). RESULTS: We found no differences in dietary intake or clinically significant medical illness between hypocholesterolemic and control subjects. In the hypocholesterolemic subgroup, we found an increased frequency of the apo E2 allele (epsilon 2) and a decreased frequency of the apo E4 allele (epsilon 4); the frequencies of the epsilon 2, epsilon 3, and epsilon 4 alleles were 33.3%, 63.3%, and 3.3%, respectively. The corresponding apo E allele frequencies in the comparison subgroup were 8.2%, 73.5%, and 18.4%, similar to those previously reported for the general population and significantly different from those found in the hypocholesterolemic subgroup (p < 0.0001). One hypocholesterolemic subject (a 46th patient) had a mutation in the apo B gene that resulted in the synthesis of a truncated species of apo B (apo B-46). CONCLUSION: Our study indicates that hypocholesterolemia in our KFHP urban population is usually not caused by diet or disease. Biochemical factors, including the increased frequency of the apo E-2 phenotype and the decreased frequency of the apo E-4 phenotype, are more important. PMID- 8238065 TI - Pseudomonas aeruginosa and Enterobacteriaceae bacteremia after biliary endoscopy: an outbreak investigation using DNA macrorestriction analysis. AB - PURPOSE: An outbreak of gram-negative bacteremia in patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) was investigated to determine the sources of infection and to control transmission. PATIENTS, METHODS, AND RESULTS: The incidence of post-ERCP bacteremia increased from 1.6% (60 of 3,696) procedures to 3.6% (53 of 1,454) procedures (relative risk 2.3, p < 0.0001) after endoscopes were processed in a new automated disinfector. Bacteremia involved nine species of Pseudomonas and Enterobacteriaceae, which were also isolated from processed endoscopes. Seven epidemic strains with highly related genomic macrorestriction profiles each infected 2 or more patients, accounting for 29 (55%) episodes of post-ERCP bacteremia. Strains recovered from endoscopes and from the disinfector were associated with 22 (42%) and 5 (9%) bacteremic episodes respectively. Effective endoscope disinfection was achieved by cleansing and disinfection of a blind channel not processed in the disinfector, additional isopropanol-air flush of all channels, and auto disinfection of the disinfector. In the following period, the incidence of post ERCP bacteremia returned to the pre-epidemic rate (1.7%, p = 0.0001). CONCLUSION: Bacterial genome fingerprinting by macrorestriction analysis enabled delineation of a multi-strain outbreak of post-ERCP bacteremia. Cross-contamination, and to a lesser extent, common-source contamination, appeared related to inadequate disinfection of endoscopes processed in an automated disinfector. PMID- 8238066 TI - Past infection by Chlamydia pneumoniae strain TWAR and asymptomatic carotid atherosclerosis. Atherosclerosis Risk in Communities (ARIC) Study Investigators. AB - PURPOSE: To determine whether past infection by Chlamydia pneumoniae strain TWAR is associated with asymptomatic atherosclerosis. Previous studies have linked this organism with symptomatic coronary heart disease. SUBJECTS AND METHODS: Between 1986 and 1989, 15,800 men and women aged 45 to 64 years were examined as part of the Atherosclerosis Risk in Communities Study, a prospective cohort study of atherosclerosis being conducted in 4 United States communities. The examination included B-mode ultrasonography of the carotid arteries and an assessment of cardiovascular disease risk factors. Carotid wall thickening (blood intima to medial-adventitial interface) in the absence of clinical cardiovascular disease was considered evidence of asymptomatic atherosclerosis. In 1991, IgG antibody titers to TWAR were assayed by microimmunofluorescence in stored sera from 326 case-control pairs matched by age group, race, sex, examination period, and field center. A titer of 1:8 or higher was considered a positive TWAR antibody response. RESULTS: Seventy-three percent of atherosclerosis cases had serologic evidence of past TWAR infection versus 63% of controls (matched odds ratio 1.76; 95% confidence interval, 1.21 to 2.57). After adjustment for age, hypertension, diabetes, cigarette smoking, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and education, the odds ratio for atherosclerosis was essentially unchanged at 2.00 (95% confidence interval, 1.19 to 3.35). The association was stronger for individuals aged 45 to 54 years than for those aged 55 to 64 years. CONCLUSION: There was a significant cross sectional association between past TWAR infection and asymptomatic atherosclerosis. This organism may be a contributor to the pathogenesis of atherosclerosis. PMID- 8238067 TI - Fever of nosocomial origin: etiology, risk factors, and outcomes. AB - OBJECTIVES: To identify the etiologies and risk factors associated with fever of nosocomial origin and compare the outcomes of patients with and without nosocomial fever. DESIGN: A prospective study with a case-control component. SETTING: The three medical services of a university hospital. PATIENTS AND METHODS: We evaluated 100 patients with fever of nosocomial origin. Etiologies were assigned by the consensus of three independent reviewers using Centers for Disease Control guidelines. Predictors of bacterial etiology were identified using logistic regression methods. Controls matched for age (+/- 5 years), sex, and comorbidity were identified for 65 of the nosocomial fever cases. Cases and controls were compared with conditional logistic regression to identify risk factors for fever of nosocomial origin. Cases and controls were followed at 1 month to compare length of stay, mortality, and hospital readmission rates. MEASUREMENTS AND MAIN RESULTS: An etiology was assigned in 81 cases: 51 had bacterial infection, 5 had nonbacterial infection, and 25 had a noninfectious etiology. Multivariate predictors of bacterial infection were the presence of diabetes mellitus (odds ratio [OR] = 5.74; 95% confidence interval of the odds ratio [CI] = 1.61 to 20.40), length of stay to fever onset after admission greater than 10 days (OR = 3.98, CI = 1.04 to 15.22), maximum temperature greater than 38.7 degrees C (OR = 3.37, CI = 1.28 to 8.88), and white blood cell count greater than 10 x 10(9)/L (OR = 4.64, CI = 1.68 to 12.77). Comparison of cases and controls indicated that patients with nosocomial fever had more invasive procedures in the 72 hours preceding enrollment in the study than controls (OR = 1.46, CI = 1.03 to 2.08). Although cases were hospitalized almost twice as long as controls (21.5 versus 12.5 days; p < 0.0001), neither 30-day mortality nor hospital readmission rates differed between cases and controls. CONCLUSION: Nosocomial fever often does not represent nosocomial infection and may constitute a wide variety of other conditions. Since bacterial etiology of fever is a major concern, clinical and laboratory findings can be used to predict patients at high risk for an infectious process. A reduction in the number of invasive procedures may decrease the incidence of nosocomial fever. New-onset nosocomial fever leads to an increase in length of hospital stay but is not a significant predictor of mortality. PMID- 8238068 TI - Spontaneous variability of ventricular function in patients with chronic heart failure. The Western Enoximone Study Group and the REFLECT Investigators. AB - PURPOSE: The spontaneous variation of the left and right ventricular ejection fractions (LVEF and RVEF) was evaluated in patients with chronic heart failure receiving only digoxin and diuretics over a 12-week interval. PATIENTS AND METHODS: Fifty-one patients with stable heart failure were studied with radionuclide angiography. A baseline evaluation and a 12-week follow-up study were performed. Heart failure therapy consisted of digoxin and diuretics alone during this time. RESULTS: The mean baseline LVEF (n = 51) was 27.2 +/- 9.5 (range: 7 to 50) and the LVEF after 12 weeks was 27.6 +/- 9.7 (range: 11 to 53; p = NS versus baseline). Mean RVEF (n = 19) was 31.9 +/- 11.3 at baseline and 30.3 +/- 11.3 (range: 14 to 50; p = NS versus baseline) after 12 weeks. Although there was no significant change in mean LVEF or RVEF in this group of patients, individual patients demonstrated considerable spontaneous variation. Individual LVEF values changed from 0 to 26 ejection fraction percentage points (mean of individual changes = 5.6 +/- 5.5). Individual RVEF determinations over the 12 week period varied by 0 to 15 percentage units (mean = 5.6 +/- 4.9). Thirty-five percent of patients had an absolute change in LVEF greater than 5 and 37% of patients had an absolute change of RVEF greater than 5. Even after deletion of the two worst outliers from the LVEF and RVEF data, a change in LVEF greater than 13 and a change in RVEF greater than 11% units were necessary to exclude spontaneous variation as a likely cause for the observed changes (95% confidence limits). No relationship between a change in the individual patient's LVEF or RVEF was found when these values were compared with exercise time, systolic or diastolic blood pressure, heart rate, or degree of baseline left or right ventricular dysfunction. CONCLUSION: In patients with heart failure, large (greater than 5) spontaneous changes in LVEF and RVEF may be seen in over one third of patients during a 12-week period. This variability should be considered when the ejection fraction is used as an index of improved or worsened cardiac function. The use of the LVEF and RVEF to assess interventions or therapy for heart failure should be interpreted with caution. PMID- 8238069 TI - Sickle myonecrosis revisited. AB - This review focuses on the classification of sickle myonecrosis in the context of sickle cell vaso-occlusive crisis. Further, the potential and novel use of magnetic resonance imaging to further clarify the nature of sickle crisis is discussed. PMID- 8238070 TI - The war within: 3. PMID- 8238071 TI - Hypothyroidism and enlarging thyroid mass in a young woman. PMID- 8238072 TI - Early manifestation of thrombotic thrombocytopenic purpura. PMID- 8238073 TI - Primary T-cell lymphoma of muscle in a patient infected with human immunodeficiency virus. PMID- 8238074 TI - Transient thyroiditis due to surgical trauma. PMID- 8238075 TI - Lovastatin-associated sleep and mood disturbances. PMID- 8238076 TI - Prostaglandin F2 alpha in the treatment of vinca alkaloid-induced ileus. PMID- 8238077 TI - Essential hypertension genetics, HLA associations, and heterozygous hemochromatosis. PMID- 8238078 TI - Could anti-beta 2 glycoprotein I antibodies potentiate antiphospholipid autoantibody activity? PMID- 8238079 TI - Effect of subclinical and overt hypothyroidism on the plasma lipid profile. PMID- 8238080 TI - Physician responsibility in driver assessment. AB - This study describes the present physician practice of counseling patients about driving and reporting potentially unsafe drivers. A survey was sent to all licensed general practitioners, internists, ophthalmologists, neurologists, and neurosurgeons in Connecticut. Fifty-nine percent (2,049/3,450) of eligible physicians responded to the questionnaire. Seventy-seven percent of responding physicians stated that they discussed driving with their patients and 14% reported patients to the Department of Motor Vehicles. Specialists were more likely to discuss driving. Seventy-four percent of physicians thought that after a certain age individuals should be screened for their ability to drive. Overall, 59% of physicians thought it was their responsibility to report unsafe drivers to the Department of Motor Vehicles and the primary reasons for reporting were that driver safety was a public health issue and that potentially unsafe drivers posed a risk to themselves or to others. It was concluded that physicians recognize that driving safety is an important public health issue that needs better-defined legal and medical guidelines. PMID- 8238081 TI - Insulin resistance and hypertension: glucose intolerance, hyperinsulinemia, and elevated free fatty acids in the lean spontaneously hypertensive rat. AB - Recent evidence suggests that insulin resistance may be a feature of essential hypertension in humans. However, there is some dispute over suitable animal models. To clarify these issues, we performed oral glucose tolerance tests in lean spontaneously hypertensive rats (SHR) as well as in age-matched normotensive rats from the parent Wistar-Kyoto (WKY) strain. In response to feeding, SHR were significantly hyperglycemic and hyperinsulinemic compared to WKY. In addition, free fatty acids were significantly higher throughout the oral glucose tolerance tests in SHR compared to WKY. Furthermore, this apparent insulin resistance occurred despite the fact that SHR were significantly leaner than age-matched WKY. When growth curves were compared for the two strains fed ad libitum, both SHR and WKY gained weight appropriately during the period of observation, although SHR were significantly lighter throughout. It is concluded that SHR express insulin resistance in terms of glucose and fatty acid metabolism, and therefore are a suitable model for insulin resistance and essential hypertension in non-obese humans. Further studies are needed to clarify the pathophysiologic defects leading to insulin resistance in these animals. PMID- 8238082 TI - Resource utilization in treatment of diabetic ketoacidosis in adults. AB - This study defines determinants of cost of treatment of diabetic ketoacidosis. A chart review for 92 cases of ketoacidosis from 1988 to 1992 in 40 females and 25 males (age range 18 to 81) showed a length of stay of 5.7 +/- 4.9 d. Length of stay did not correlate with the level of nursing care or any other component of the initial order set. Length of stay was shorter in cases managed by diabetologists. Length of stay was inversely proportional to the interval from arrival to the first shot of intermediate or long-acting insulin. Length of stay was longer in subjects with a positive bacterial culture (mean, 9.1 d) and was longer in subjects who arrived in the evening. There was a female predominance in total and recurrent cases of ketoacidosis. Female patients received fewer educational sessions than males. The grade of acidosis affected the duration of acidemia, but the grade of acidosis, APACHE scores, and admission lab values did not correlate with length of stay. The use of an intensive care unit (ICU) included more testing and expense without uniform clear benefit. Optimal transition from intensive to routine management includes resumption of long acting insulins as soon as possible. Optimization of hospital care and reduction of incidence of ketoacidosis in females would have a marked effect on health care costs. PMID- 8238083 TI - Diagnosis of mixed acid-base disorders in diabetic ketoacidosis. AB - In diabetic ketoacidosis, a mixed acid-base disorder is suggested when the anion gap increase (delta AG) does not equal the bicarbonate decrease (delta HCO3), or when the delta AG/delta HCO3 ratio does not equal 1.0. It is widely assumed that delta AG/delta HCO3 is significantly different from 1.0 when it is less than 0.8 or greater than 1.2. The validity of these ratio limits were examined by analyzing a normal control group of 68 subjects and 27 diabetic ketoacidosis admissions that had no evidence of mixed disorders. In the 27 ketoacidosis admissions, regression analysis showed that delta AG was predicted to equal delta HCO3, as expected in pure anion gap acidosis: delta AG = 1.0 delta HCO3 (r = 0.744, p < 0.001). It was found that delta AG is significantly different from delta HCO3 when they differ by more than 8 mEq/L, and equivalently, delta AG/delta HCO3 is significantly different from 1.0 when it is less than (1.0 - 8/delta HCO3) or greater than (1.0 + 8/delta HCO3). These criteria from regression analysis suggested that 4% of the 27 pure anion gap acidoses, and 3% of the control group, had mixed disorders. In contrast, the ratio limits of 0.8 and 1.2 suggested 56% of the pure anion gap acidoses, and 94% of the control group, had mixed disorders. It was concluded that mixed disorders are overdiagnosed by the ratio limits of 0.8 and 1.2. Mixed disorders are more accurately detected by noting whether delta AG and delta HCO3 differ by more than 8 mEq/L. PMID- 8238084 TI - Case report: profound neurobehavioral deficits in an oil field worker overcome by hydrogen sulfide. AB - A 24-year-old oil well tester was rendered semiconscious by hydrogen sulfide (H2S). He received oxygen and was hospitalized but released in 30 minutes. The next day, nausea, vomiting, diarrhea, and incontinence of urine and stool led to rehospitalization. These problems and leg shaking, dizziness, sweating, trouble sleeping, and nightmares prevented his return to work. A physical examination, chest x-ray, and pulmonary function tests were normal 39 months after the episode but vibration sense was diminished. Two choice visual reaction times were delayed. Balance was highly abnormal (5 to 6 cm/sec) with eyes closed. Blink reflex latency was slow (R-1 17.5 msec versus normal 14.3 msec). Numbers written on finger tips were not recognized. Verbal and visual recall were impaired but overlearned memory was intact. Cognitive functions measured by Culture Fair, block design, and digit symbol were impaired. Perceptual motor was slow. Scores for confusion, tension-anxiety, depression, and fatigue were elevated and vigor was reduced. Forty-nine months after exposure his reaction time, sway speed, and color vision had not improved. His recall and his cognitive, constructional, and psychomotor speeds had improved but remained abnormal. These deficits are most likely due to H2S. Similar testing of other survivors is recommended. PMID- 8238085 TI - Case report: a case of multiple coronary artery to left ventricular communications. AB - A 39-year-old man with anginal pain had multiple coronary artery to left ventricular communications. His electrocardiogram showed evidence of left ventricular hypertrophy, and an echocardiogram revealed a dilated left ventricle. A coronary angiogram revealed multiple coronary artery to left ventricular fistulae involving three major coronary arteries with no evidence of atherosclerotic lesions. Only 17 cases of such fistulous communications involving three major coronary arteries have been reported in the literature. It is suggested that the fistulous communications to the left ventricle was a cause of his angina pectoris, probably because of the coronary steal phenomenon. PMID- 8238086 TI - Case report: long-term remission of parathyroid cancer: possible relation to vitamin D and calcitriol therapy. AB - Recurrence of surgically treated parathyroid cancer occurs in 30% to 65% of patients and has a poor prognosis; only 1 of 29 cases remained normocalcemic more than 2 years later. No medical attempts to prevent recurrence have been reported. A 24-year-old pregnant woman whose mother died of parathyroid cancer underwent apparently successful surgery for parathyroid cancer. Serum Ca and parathyroid hormone (PTH) returned to normal levels but 3 months after surgery, although normocalcemic, the serum PTH level was elevated. The administration of vitamin D 200,000 U/month or calcitriol 0.5 microgram daily and 1 g of Ca supplementation daily, resulted in the normalization of PTH during 81 months of follow-up. On three occasions, when vitamin D or calcitriol were omitted, serum intact, C terminal, or mid-molecule PTH levels rose. Ionized and total serum Ca, creatinine, calcitriol and calcidiol levels were normal, and multiple ultrasounds of the neck remained negative after surgery. This observation suggests that serum PTH could be an early marker for the detection of recurrence in parathyroid cancer with normal serum Ca, and that suppression of PTH secretion by vitamin D or calcitriol could avert or delay the progression of recurrence. Additional trials with calcitriol in operated normocalcemic parathyroid cancer with an elevated serum PTH level is recommended. PMID- 8238087 TI - Case report: hypercalcemia in a patient with AIDS and Pneumocystis carinii pneumonia. AB - Pneumocystis carinii infection is commonly seen in patients infected with HIV, and there is evidence of macrophage involvement in the disease process. Macrophage dysfunction can result in abnormal vitamin D metabolism as is often seen in a granulomatous disease such as sarcoidosis. This article describes a patient with AIDS who had P. carinii pneumonia and hypercalcemia and had elevated 1,25-dihydroxyvitamin D levels, the first such reported case in the literature. There was no other evidence of a granulomatous disease such as sarcoidosis or tuberculosis to account for this. It is suggested that the increase in 1,25 dihydroxyvitamin D level was secondary to P. carinii induced macrophage dysfunction. As the patient's P. carinii pneumonia resolved, his 1,25 dihydroxyvitamin D level normalized along with the resolution of hypercalcemia. PMID- 8238088 TI - Case report: pseudoephedrine-associated thyroid storm: thyroid hormone catecholamine interactions. AB - Thyroid storm is an increasingly rare, life-threatening manifestation of thyrotoxicosis that can be precipitated by many physiologic stressors. The exact mechanism by which thyrotoxicosis decompensates into thyroid storm in certain individuals is unknown. These individuals may have an enhanced cellular sensitivity to either catecholamines or thyroid hormone. Serum levels of catecholamines are usually low or normal in thyrotoxic individuals. Ingestion of sympathomimetic drugs in thyrotoxic individuals may precipitate thyroid storm. This article presents an unusual case of Graves' disease that decompensated into thyroid storm after the ingestion of an over-the-counter cold medication containing pseudoephedrine. PMID- 8238089 TI - Case report: amelioration of insulin resistance in diabetes with dehydroepiandrosterone. AB - In hyperandrogenic females, the ratio of dehydroepiandrosterone (DHEA) to testosterone may be an important determinant of insulin sensitivity. This study involved changes in insulin sensitivity and glucose metabolism with therapeutic manipulation of DHEA (S)/testosterone in a female patient with non-insulin dependent diabetes and hyperandrogenism. Therapeutic intervention included 1 month treatment with 0.25 mg dexamethasone at bedtime and 1-month dexamethasone + DHEA. Insulin sensitivity and glucose tolerance were assessed before and after each treatment regimen by examining: 1) fasting and oral glucose tolerance test glucose and insulin levels, 2) hypoglycemic response to intravenous insulin, and 3) erythrocyte insulin receptor binding. With dexamethasone alone, DHEAS, testosterone, and their ratio were reduced with a concomitant increase (30%) in oral glucose tolerance test insulin levels and a decrease (33%) in erythrocyte insulin binding. With DHEA + dexamethasone, the ratio of DHEAS/testosterone increased 16-fold along with a marked improvement in insulin sensitivity, as determined by a more than 30% reduction in fasting and oral glucose tolerance test insulin levels, a threefold stimulation of the rate of glucose disappearance with intravenous insulin, and a 30% increase in insulin binding. DHEA improved insulin sensitivity and reduced fasting and oral glucose tolerance test glucose levels and ameliorated the diabetic state. The ratio of DHEAS/testosterone is an important regulator of insulin sensitivity and glucose tolerance and that DHEA therapy may be beneficial in the treatment of certain forms of insulin resistance. PMID- 8238090 TI - Role of the S-layer proteins of Campylobacter fetus in serum-resistance and antigenic variation: a model of bacterial pathogenesis. AB - Campylobacter fetus are microaerophilic gram-negative bacteria that are pathogens of animals and humans. These organisms possess paracrystalline surface (S-) layers, composed of acidic high molecular weight proteins. C. fetus strains possessing S-layers are resistant to C3b binding, which explains both serum and phagocytosis-resistance. C. fetus strains also can vary the subunit protein size, crystalline structure, and antigenicity of the S-layer it expresses. Therefore, its S-layer permits C. fetus to resist complement and antibodies, two of the key defenses against extracellular pathogens. C. fetus possesses several full-length genes encoding S-layer proteins with both conserved and divergent sequences, which permits gene rearrangement and antigenic variation. PMID- 8238091 TI - Perioperative prophylaxis of venous thromboembolism. AB - Venous thromboembolism is an important cause of morbidity and mortality in hospitalized patients, causing 100,000 to 200,000 deaths per year in the United States. Patients undergoing surgery are at the highest risk of venous thromboembolism. The magnitude of this risk in a patient depends on the surgical procedure performed and the presence of other risk factors that predispose to venous thromboembolism. The clinical diagnosis of both deep vein thrombosis and pulmonary embolism is notoriously inaccurate. Furthermore, two thirds of all fatal pulmonary emboli cause death within 30 minutes of the embolic episode, leaving little time for diagnostic work-up and effective treatment. Prophylactic treatment for prevention of venous thromboembolism is therefore important in these patients, and several effective mechanical and chemical methods for this purpose are available. The pathogenesis of deep vein thrombosis in the surgical patients, the predisposing risk factors, and the available prophylactic modalities are discussed in this article. Recommendations for the use of various approaches in different risk categories are provided. PMID- 8238092 TI - Insulin resistance in hypertension: a focused review. AB - The associations between insulin resistance, hyperinsulinemia, and hypertension are well recognized. The insulin resistance and hyperinsulinemia associated with hypertension is the result of increased renal tubular sodium reabsorption, increased sympathetic nervous system activity, and increased arterial wall smooth muscle reactivity. In insulin resistant states, intracellular calcium and sodium accumulation is thought to be the fundamental underlying abnormality. These is evidence that hyperinsulinemia is an independent risk factor for coronary artery disease. Therefore, therapeutic considerations in patients with insulin resistance should include nonpharmacologic methods for increasing insulin sensitivity as well as avoiding the deleterious effects on insulin action and lipid metabolism of various antihypertensive medications. Of the currently available antihypertensives therapies, the angiotensin-converting enzyme inhibitors, the alpha antagonists, and the calcium channel blockers are the most rational choices for treating patients with hypertension with evidence of coexisting insulin resistance. PMID- 8238093 TI - Always read medication labels. PMID- 8238094 TI - 24-hour pharmacy service a must. PMID- 8238095 TI - Taking the risk to care. PMID- 8238096 TI - Taking the risk to care. PMID- 8238097 TI - Bottom-line issues in home care. PMID- 8238098 TI - "What am I going to do now? PMID- 8238099 TI - Abdominal trauma: putting the pieces together. PMID- 8238100 TI - Home before dark: one nurse's personal experience with diabetic neuropathy. PMID- 8238101 TI - Back to basics: acidosis, alkalosis, and ABGs. PMID- 8238102 TI - Retraining the bladder after catheterization. PMID- 8238103 TI - Caring for the cardiac spouse. PMID- 8238104 TI - Venous air embolism: keeping your patient out of danger. PMID- 8238105 TI - The best buy in health care. PMID- 8238106 TI - The view from the bed. PMID- 8238107 TI - Progress notes on women's health. PMID- 8238108 TI - Speaking of sex. PMID- 8238109 TI - Maternal morbidity and mortality in 442 pregnancies with hemolysis, elevated liver enzymes, and low platelets (HELLP syndrome) AB - OBJECTIVE: Our purpose was to describe the incidence and effects of serious obstetric complications on maternal outcome in pregnancies complicated by HELLP syndrome. STUDY DESIGN: A prospective cohort study was performed on 442 pregnancies with HELLP syndrome managed at this center from August 1977 through July 1992. RESULTS: Of 437 women who had 442 pregnancies with HELLP syndrome; 309 (70%) of the cases occurred ante partum and 133 (30%) post partum; 149 (11%) developed at < 27 weeks and 80 (18%) at term. Maternal mortality was 1.1% (five patients). Serious maternal morbidity included disseminated intravascular coagulation (21%), abruptio placentae (16%), acute renal failure (7.7%), pulmonary edema (6%), subcapsular liver hematoma (0.9%), and retinal detachment (0.9%). Fifty-five percent of patients required transfusions with blood or blood products, and 2% required laparotomies for major intraabdominal bleeding. Abruptio placentae was strongly correlated with the development of disseminated intravascular coagulation (p < 0.0001), acute renal failure (p < 0.001), and pulmonary edema (p < 0.01). Moreover, there was a strong association between pulmonary edema and acute renal failure (p < 0.0001). There were no differences in laboratory findings between HELLP syndrome before and after delivery; however, women with postpartum HELLP syndrome had significantly higher incidences of pulmonary edema and renal failure. CONCLUSION: HELLP syndrome is associated with serious maternal morbidity, especially when it arises in the postpartum period. PMID- 8238110 TI - Peroxide induces vasoconstriction in the human placenta by stimulating thromboxane. AB - OBJECTIVE: Placental lipid peroxides and thromboxane are abnormally increased in preeclampsia. Thromboxane is a potent vasoconstrictor of the placental vasculature. Peroxides stimulate cyclooxygenase (prostaglandin H synthase), and thereby could increase thromboxane, to cause vasoconstriction in the placenta. This study was performed to test the hypothesis that peroxides would produce vasoconstriction in the human placenta by stimulating thromboxane production. STUDY DESIGN: Isolated human placental cotyledons were perfused for 20-minute intervals with 100 mumol/L t-butyl hydroperoxide alone, and during and after perfusion with low-dose aspirin (5 x 10(-5) mol/L) (n = 6) or the thromboxane receptor blocker SQ 29,548 (n = 2). Krebs-Ringer-bicarbonate buffer gassed with 95% oxygen and 5% carbon dioxide was used for the perfusion buffer. Perfusion pressure was monitored continuously, and effluent flow rates were measured during each experimental treatment. Maternal and fetal effluent samples were analyzed for thromboxane B2 and 6-keto-prostaglandin F1 alpha. RESULTS: Compared with control Krebs-Ringer-bicarbonate buffer perfusion, peroxide perfusion significantly increased (p < 0.05) vascular resistance (14 +/- 2 vs 25 +/- 3 mm Hg.min/ml, mean +/- SE, respectively), thromboxane B2 secretion (fetal 0.20 +/- 0.04 vs 1.65 +/- 0.26 ng/min, maternal 4.8 +/- 1.5 vs 8.1 +/- 2.1 ng/min) and 6 keto-prostaglandin F1 alpha secretion (fetal 21 +/- 5 vs 60 +/- 1.8 pg/min, maternal nondetectable). Peroxide perfusion increased the thromboxane B2/6-keto prostaglandin F1 alpha ratio threefold on the fetal side. Subsequent perfusion with aspirin significantly blocked the peroxide-induced vasoconstriction (13 +/- 1 mm Hg.min/ml during aspirin + peroxide) and the peroxide-induced increase in the secretion of thromboxane B2 (fetal 0.52 +/- 0.12 ng/min, maternal 2.0 +/- 0.3 ng/min) and 6-keto-prostaglandin F1 alpha (fetal 30 +/- 8 pg/min). After perfusion with aspirin the thromboxane B2/6-keto-prostaglandin F1 alpha ratio declined. When the thromboxane receptor blocker was used instead of aspirin, maternal and fetal secretion rates of thromboxane were still significantly increased by peroxide perfusion, but there was no change in perfusion pressure or vascular resistance. CONCLUSIONS: (1) Peroxide induces placental vasoconstriction coincident with increased secretion of thromboxane; (2) low-dose aspirin blocks both increased thromboxane secretion and vasoconstriction, whereas a thromboxane receptor antagonist allows increased thromboxane secretion but prevents peroxide induced vasoconstriction; (3) therefore, peroxide induces vasoconstriction by stimulating thromboxane synthesis. PMID- 8238111 TI - The risk of second-trimester amniocentesis in twin gestations: a case-control study. AB - OBJECTIVE: Pregnancy outcomes in patients with twin pregnancy undergoing second trimester amniocentesis for fetal karyotype assessment were compared in a case control study with twin pregnancies undergoing routine ultrasonographic studies at similar gestational ages. STUDY DESIGN: All spontaneous and induced twin gestations that underwent ultrasonographic examination between 14 and 20 weeks were compiled for the period January 1987 through January 1992. Patients having undergone multifetal reduction or chorionic villous sampling and those with fetal anatomic or chromosomal anomalies, discordant growth (> 20%), death, or a monoamniotic sac detected at ultrasonography were excluded. RESULTS: The mean (+/ SD) maternal age was significantly higher among the 101 cases than among the 108 controls (35.2 +/- 3.5 vs 30.4 +/- 5.3 years, respectively; p < 0.01). No differences were noted in gravidity, parity, number of prior spontaneous losses, or gestational age at ultrasonography between the two groups. The fetal loss rate was similar among cases and controls (seven of 202 [3.5%] vs seven of 216 [3.2%], relative risk 1.07, 95% confidence intervals 0.3 to 3.5). No losses occurred within 3 weeks of the procedure. Gestational age at delivery, birth weight, mean Apgar scores at 1 and 5 minutes, and length of neonatal stay were not significantly different between cases and controls. CONCLUSIONS: Second-trimester amniocentesis in twin pregnancies is apparently not associated with excess pregnancy loss. PMID- 8238112 TI - Tissue-specific mosaicism among fetuses with prenatally diagnosed diaphragmatic hernia. AB - OBJECTIVE: Our purpose was to determine if cytogenetic discrepancies between fetal blood and amniotic fluid are present in fetuses with prenatally diagnosed diaphragmatic hernia. STUDY DESIGN: Chromosome analysis was performed on 15 fetuses with prenatally diagnosed diaphragmatic hernia. Fourteen had both amniotic fluid and fetal blood studies. One fetus had an amniocentesis followed by postnatal skin and peripheral lymphocyte chromosome analysis. RESULTS: In one fetus with a normal karyotype on fetal blood, amniotic fluid mosaicism for a supernumerary isochromosome 12p was identified. Another fetus had normal amniotic fluid chromosome analysis but was diagnosed with mosaic isochromosome 12p on skin biopsy after birth. Concordant aneuploidy in both fetal blood and amniocytes was found in five pregnancies (three with trisomy 18, one with an unbalanced translocation, and one with mosaic supernumerary isochromosome 12p). Eight fetuses had normal karyotypes. CONCLUSION: Because diaphragmatic hernia is a common component of mosaic isochromosome 12p syndrome and this chromosome abnormality is predominantly found in fibroblasts but not lymphocytes, an amniocentesis may be more accurate than fetal blood sampling in defining the true fetal chromosome status when diaphragmatic hernia is detected prenatally. PMID- 8238113 TI - Cook obstetrics and gynecology catheter multicenter chorionic villus sampling trial: comparison of birth defects with expected rates. AB - OBJECTIVE: The null hypothesis was that offspring of women undergoing first trimester chorionic villus sampling do not experience a rate of birth defects exceeding background rates. STUDY DESIGN: Follow-up information regarding major malformations was prospectively sought on offspring of 4105 women undergoing first-trimester chorionic villus sampling from nine centers participating in a collaborative study with the Cook obstetrics and gynecology catheter. These data were compared with data from the Collaborative Perinatal Project and other registries. RESULTS: A total of 84 offspring with major malformations was identified (2.36%). Compared with background rates, there was no increase in the incidence of total malformations or specific malformations (including limb reduction defects) in the subjects. One institution experienced all three limb reduction defects in this series; the probability of this occurring by chance alone is < 1%. CONCLUSION: Chorionic villus sampling was not found to result in an increase in major birth defects or in specific categories of birth defects in this series. PMID- 8238114 TI - Effects of hydralazine on pregnant rats and their fetuses. AB - OBJECTIVE: We conducted this study to assess potential teratogenic or fetotoxic effects of in utero hydralazine exposure in the rat. STUDY DESIGN: Pregnant rats were treated with hydralazine (0, 15, or 30 mg/kg/day) by either gavage or subcutaneous injection during gestation days 8 through 19, inclusively. Surviving animals were killed, uterine contents examined, and fetuses evaluated by Wilson's method for teratologic examination. RESULTS: Oral administration did not affect maternal weight gain or fetal or placental weights. Subcutaneous administration decreased fetal and placental weights but only in dams given the highest dose subcutaneously; significantly increased maternal mortality and reduced maternal weight gain were also observed. There was no increased incidence of fetal anomalies in any group. CONCLUSION: Hydralazine does not have teratogenic or fetopathic effects when given to rats in dosages that are nontoxic to the mother. PMID- 8238115 TI - The risk of sexually transmitted diseases in human immunodeficiency virus infected parturients. AB - OBJECTIVE: We assessed the prevalence of and defined the relationship between other sexually transmitted diseases and human immunodeficiency virus infection. STUDY DESIGN: We performed a case-control study among 121 human immunodeficiency virus-infected and 222 randomly selected seronegative parturient women. These women were identified from a prenatal population undergoing routine voluntary antibody screening in a large urban hospital in the southeastern United States. RESULTS: During the 24-month study period, 16,868 women consented to human immunodeficiency virus antibody screening; 121 (7.2/1000) were infected with human immunodeficiency virus. Cases were significantly more likely than controls to be infected with at least one sexually transmitted disease during pregnancy (48% vs 21%; odds ratio 3.4, 95% confidence interval 2.1 to 5.7). The prevalence of Chlamydia trachomatis and hepatitis B infection did not differ significantly among the groups. Cases were significantly more likely than controls to be infected with Treponema pallidum (29% vs 4%; odds ratio 9.6, 95% confidence interval 4.2 to 22.4). This relationship persisted after we controlled for confounding risk factors (odds ratio 9.2, 95% confidence interval 2.1 to 13.3). In addition, cases were significantly more likely than controls to be infected with Neisseria gonorrhoeae (17.2% vs 4%; odds ratio 5.2, 95% confidence interval 2.1 to 13.3). This relationship also persisted after we controlled for confounders (odds ratio 3.7, 95% confidence interval 1.4 to 10.0). CONCLUSION: Human immunodeficiency virus-infected parturient women in our center are at substantial risk of having other sexually transmitted diseases. PMID- 8238116 TI - Amniotic fluid interleukin-6 levels correlate with histologic chorioamnionitis and amniotic fluid cultures in patients in premature labor with intact membranes. AB - OBJECTIVES: Our purpose was (1) to determine the frequency of intraamniotic and extraamniotic intrauterine infection in patients with premature labor and intact membranes and (2) to determine if intrauterine infection is associated with elevated amniotic fluid interleukin-6 levels. STUDY DESIGN: Amniocentesis was performed on 57 patients in preterm labor and 201 controls at various gestational ages without labor and at term with labor. The amniotic fluid was evaluated with gram stain, cultures, and an enzyme-linked immunosorbent assay specific for interleukin-6. Placentas from study patients (n = 52) and term controls (n = 120) were analyzed. RESULTS: The frequency of positive amniotic fluid cultures (intraamniotic intrauterine infection) was 10 of 57 (18%) in the preterm labor group and zero of 201 for controls. Histologic chorioamnionitis (extraamniotic intrauterine infection) was present in 21 of 24 (88%) of patients in preterm labor that failed tocolysis and 28 of 120 (23%) of term laboring controls. An amniotic fluid interleukin-6 level of > or = 600 pg/ml was 100% sensitive and 89% specific (positive predictive value 85%, negative predictive value 100%) for the identification of intrauterine infection. CONCLUSION: Interleukin-6 is a sensitive and specific marker for the identification of both intraamniotic and extraamniotic intrauterine infection in patients in preterm labor with intact membranes. PMID- 8238117 TI - Accelerated pulmonary maturation from preterm premature rupture of membranes: a myth. AB - OBJECTIVES: It is widely believed that premature rupture of membranes accelerates fetal pulmonary maturity. The purpose of our study was to determine the duration of premature rupture of the membranes required to achieve this effect. STUDY DESIGN: Retrospective analysis of our database yielded a group of 1395 patients who were delivered between 24 and 35 weeks' gestation and for whom we had complete data. The frequencies of premature rupture of the membranes and respiratory distress syndrome by each gestational week were analyzed with a log linear multiway contingency table analysis. Because gestational age was based on pediatric examination and was therefore somewhat subjective, birth weight was used to confirm results. Additional factors related to respiratory distress syndrome were considered in stepwise discriminant analysis. Results were further verified by the 1980 National Natality Survey data set. RESULTS: When we controlled for either gestational age or birth weight, there was no significant difference in the frequency of respiratory distress syndrome related to premature rupture of the membranes, but there was a suggestion (p < 0.08) that respiratory distress syndrome was actually more frequent after premature rupture of the membranes. Stepwise discriminant analysis revealed that gestational age, birth weight, race, sex, and Apgar score at 1 minute were all more important determinants than duration of premature rupture of the membranes. Duration of premature rupture of the membranes was associated with an increased risk of respiratory distress syndrome. Amnionitis was found to be highly related to the duration of premature rupture of the membranes. The incidence of amnionitis significantly increased 24 hours after premature rupture of the membranes occurred. A multiway frequency contingency table of the National Natality Survey data showed a significant increase in respiratory distress syndrome in association with premature rupture of the membranes. CONCLUSIONS: Pulmonary maturation continues but is not accelerated after premature rupture of the membranes. In fact, there is a strong suggestion that premature rupture of the membranes actually increases the risk of respiratory distress syndrome at a given gestational age. PMID- 8238118 TI - Elective cesarean section improves outcomes of neonates with gastroschisis. AB - OBJECTIVE: Our objective was to compare neonatal postoperative morbidity for the neonate with prenatally diagnosed gastroschisis delivered vaginally with that for the perinate undergoing elective cesarean at or before the onset of labor. STUDY DESIGN: Retrospective maternal and neonatal data were obtained by chart review on 22 neonates prenatally diagnosed with gastroschisis who underwent operative closure of the ventral wall defect between 1987 and 1991 at Loma Linda University Medical Center. Perioperative data and postoperative courses were compared between 12 infants who underwent labor with vaginal delivery and 10 infants who were delivered by elective cesarean section at or before the onset of labor. RESULTS: Neonatal transports and significant bowel edema were more likely (p < 0.05) in the vaginal delivery group. The elective cesarean section infants had less sepsis (p < 0.05), fewer hospital days (p < 0.01) and parenteral nutrition days (p < 0.01), and shorter time to enteral feedings (p < 0.01). CONCLUSIONS: Elective cesarean section at or before the onset of labor may benefit the fetus with gastroschisis, compared with undergoing labor and vaginal delivery. PMID- 8238119 TI - Blunt abdominal trauma: are there any predictive factors for abruptio placentae or maternal-fetal distress? AB - OBJECTIVES: Our objectives were to determine the incidence of abruptio placentae and fetal distress in pregnant women with noncatastrophic blunt abdominal trauma and to determine the utility of historical factors, clinical presentation, coagulation profile, and fetal monitoring in predicting fetal and maternal morbidity. STUDY DESIGN: We reviewed 233 consecutive hospitalizations for noncatastrophic blunt abdominal trauma. Outcome variables included abruptio placentae, fetal distress, preterm birth, and abnormal laboratory values. RESULTS: Preterm delivery (< 34 weeks) occurred within 1 week of trauma in only two patients (< 1%). Fetal distress was diagnosed between 4 and 48 hours after observation in four women (1.7%). These four patients had frequent contractions, but none had abruptio placentae at delivery. Six patients (2.6%) had abruptio placentae. None of these had fetal distress. All had a good neonatal outcome. Coagulation studies and Kleihauer-Betke tests were not predictive of fetal or maternal morbidity. CONCLUSION: Noncatastrophic blunt abdominal trauma is associated with a low frequency of abruptio placentae, fetal distress, maternal coagulopathy, and poor neonatal outcome. Prolonged monitoring is indicated only when there is evidence of impending abruptio placentae. Kleihauer-Betke testing is necessary only for patients who are Rh negative. Coagulation profiles are not clinically helpful. Routine hospitalization beyond 4 hours is not warranted. PMID- 8238120 TI - Currently recommended oral regimens for ritodrine tocolysis result in extremely low plasma levels. AB - OBJECTIVE: Our aim was to compare plasma drug levels in patients receiving ritodrine intravenously with those in patients receiving ritodrine orally at recommended dosages. STUDY DESIGN: Plasma samples from 20 pregnant patients treated with intravenous ritodrine (50 to 300 micrograms/min), 9 patients treated with oral ritodrine only (60 to 120 mg per 24 hours), and 9 patients treated first with intravenous and subsequently with oral ritodrine were analyzed for ritodrine concentration with the use of high-performance liquid chromatography. RESULTS: Average plasma ritodrine levels of patients receiving different intravenous dosages ranged from 27.8 +/- 3.5 to 113.3 +/- 38.8 ng/ml. Levels during oral therapy ranged between 9.8 +/- 3.2 and 13.8 +/- 4.4 ng/ml. In both modes of drug delivery, concentrations were significantly correlated with doses. In patients treated first with intravenous ritodrine and subsequently with the oral form, plasma concentrations during oral therapy averaged 27.7% +/- 18.8% of those obtained during intravenous infusion. CONCLUSION: Subtherapeutic plasma concentrations might be responsible for the failure to demonstrate clinical benefits of oral ritodrine in prevention of recurrent preterm labor. A twofold to threefold increase in the maximum recommended oral dosage of ritodrine should be considered, especially for patients who had previously required relatively high intravenous infusion rates (> 100 micrograms/min). PMID- 8238121 TI - Randomized trial of oral indomethacin and terbutaline sulfate for the long-term suppression of preterm labor. AB - OBJECTIVE: Our purpose was to determine the efficacy and safety of long-term oral tocolysis with indomethacin or terbutaline sulfate. STUDY DESIGN: Seventy-one patients at 26 to 32 weeks' gestation admitted for preterm labor were prospectively randomized to receive oral indomethacin or terbutaline sulfate after successful intravenous tocolysis. Patients were monitored weekly for cervical change, maternal side effects, amniotic fluid volume, and constriction of the fetal ductus arteriosus. Patients receiving indomethacin were converted to terbutaline at 34 weeks or with the occurrence of fetal ductal constriction or oligohydramnios. RESULTS: Of 71 patients randomized six were excluded after randomization. Thirty-three patients were randomized to indomethacin and thirty two to terbutaline. There were no differences in the percentage of patients achieving 34 weeks of gestation. No differences in neonatal outcome were noted. Nine (27%) fetuses receiving indomethacin had constriction of the fetal ductus arteriosus, and 13 (38%) had oligohydramnios. Most patients on terbutaline reported beta-mimetic side effects (53%), but only one required discontinuation of therapy. CONCLUSION: Both indomethacin and terbutaline sulfate are effective tocolytics, but major fetal side effects are common with long-term indomethacin use. PMID- 8238122 TI - Randomized trial of intraumbilical vein oxytocin in midtrimester pregnancy losses. AB - OBJECTIVE: The purpose of this double-blind prospective randomized trial was to determine whether high-dose intraumbilical vein oxytocin injection shortens the third stage of labor in midtrimester pregnancy losses. STUDY DESIGN: Patients (n = 50) with spontaneous or induced midtrimester pregnancy losses (14 to 26 weeks' gestation) were randomized to receive either 100 IU of oxytocin in 20 ml of normal saline solution or 20 ml of normal saline solution alone as a placebo. The umbilical vein was injected as soon as the cord was clamped. Outcome data were collected. RESULTS: Of the 50 patients randomized, 45 completed the study. Five were excluded after randomization because of either cesarean delivery (1 patient) or en caul delivery (4 patients). Twenty-one patients received oxytocin, and 14 received placebo. Ten patients who were not injected because of technical failure were evaluated separately. There were no differences between the three groups with regard to gestational age, fetal weight, length of the third stage, blood loss, or need for operative removal of the placenta. CONCLUSION: Injection of high-dose oxytocin into the umbilical vein in second-trimester pregnancy losses does not shorten the third stage of labor or decrease the need for surgical intervention because of retained placenta. PMID- 8238123 TI - Detection of human papillomavirus deoxyribonucleic acid sequences in amniotic fluid during different periods of pregnancy. PMID- 8238124 TI - Incidence and prevalence in cerebral palsy. PMID- 8238125 TI - Cerebral palsy linkage concepts need revision. PMID- 8238126 TI - Comments on trial of labor scoring system. PMID- 8238127 TI - In vivo assessment of the early uteroplacental circulation. PMID- 8238128 TI - Auscultation of the fetal heart: how to do it. PMID- 8238129 TI - Fetal biophysical profile score. VI. Correlation with antepartum umbilical venous fetal pH. AB - OBJECTIVE: Our objective was to determine the relationship, if any, between the fetal biophysical profile score and antepartum umbilical venous pH. STUDY DESIGN: This was a prospective observational study conducted concurrently in two centers and involving two discrete high-risk groups of fetuses. Fetal biophysical profile scores were compared with umbilical venous pH values measured in blood obtained by immediate cordocentesis. A total of 493 paired observations of biophysical profile score and pH were made; 104 observations were of fetuses with intrauterine growth retardation and 389 observations were of fetuses with alloimmune anemia. RESULTS: In both data sets there was a highly significant linear correlation between biophysical profile score and umbilical venous pH. Poor biophysical profile score performance (a score of 0 of 10) was always associated with a pH < 7.20, whereas the pH was always > 7.20 when the biophysical profile score was 10 of 10. Sequenced sensitivity of short-term biophysical variables in the detection of acidemia was observed. CONCLUSION: The fetal biophysical profile score accurately predicts antepartum umbilical venous pH. PMID- 8238130 TI - Antibiotic treatment of preterm labor with intact membranes: a multicenter, randomized, double-blinded, placebo-controlled trial. AB - OBJECTIVE: Although an association between subclinical intrauterine infection and preterm birth is well established, there is conflicting evidence regarding the benefits of antibiotic administration to women in preterm labor with intact membranes. We attempted to determine the effect of ampicillin-amoxicillin and erythromycin treatment on prolongation of pregnancy, the rate of preterm birth, and neonatal morbidity in patients with preterm labor and intact membranes. STUDY DESIGN: A multicenter, randomized, double-blinded, placebo-controlled trial was designed and implemented by the Maternal-Fetal Medicine Units Network of the National Institute of Child Health and Human Development. Two hundred seventy seven women with singleton pregnancies and preterm labor with intact membranes (24 to 34 weeks) were randomly allocated to receive either antibiotics or placebos. RESULTS: Of the 2373 patients screened for participation in this study in six medical centers, 277 women were enrolled (n = 133 for antibiotics group vs n = 144 for placebo group). In each study group, 60% of patients completed all the study medications. The overall prevalence of microbial invasion of the amniotic cavity was 5.8% (14/239). No significant difference between the antibiotic group and the placebo group was found in maternal outcomes, including duration of randomization-to-delivery interval, frequency of preterm delivery (< 37 weeks), frequency of preterm premature rupture of membranes, clinical chorioamnionitis, endometritis, and number of subsequent admissions for preterm labor. Similarly, no significant difference in neonatal outcomes could be detected between the two groups including respiratory distress syndrome, bronchopulmonary dysplasia, intraventricular hemorrhage, sepsis, and admission and duration of newborn intensive special care unit hospitalization. CONCLUSION: The results of this study do not support the routine use of antibiotic administration to women in preterm labor with intact membranes. PMID- 8238131 TI - Induction versus expectant management in premature rupture of the membranes with mature amniotic fluid at 32 to 36 weeks: a randomized trial. AB - OBJECTIVE: Our objective was to compare maternal and perinatal outcomes between two management schemes for women with preterm premature rupture of the membranes and documented fetal pulmonary maturity. STUDY DESIGN: Of 164 women with preterm premature rupture of the membranes at 32 weeks to 36 weeks 6 days' gestation, 93 eligible and consenting women were randomly selected for either induction of labor (n = 46) or expectant management (n = 47). Expectant management included hospitalization, assessment for fetal heart rate abnormalities, chorioamnionitis, and labor. Digital cervical examinations were prohibited until progressive labor occurred. Follow-up was also done for the 71 women who did not participate. RESULTS: The women in the induction of labor and expectant management groups had similar demographic characteristics and gestational ages (34.1 vs 34.3 weeks). Expectant management was associated with prolonged latencies to labor, delivery, and maternal hospitalization (p < 0.001), as well as increased hospitalization of infants at 2 to 5 days after delivery (p < 0.05). These patients had increased chorioamnionitis and fetal heart rate abnormalities before labor (p = 0.01, 0.03). Infants received more frequent (p < 0.001) and prolonged antimicrobial therapy after expectant management (p = 0.003) with no reduction in proven sepsis (6.8% vs 4.4%). These latter differences were influenced by the neonatologist's concern over potential neonatal infection. CONCLUSIONS: Among women with preterm premature rupture of the membranes at 32 to 36 weeks with mature surfactant profiles, immediate induction of labor reduces the duration of hospitalization and infection in both mothers and neonates. PMID- 8238132 TI - The acute effect of cocaine exposure on pregnant human myometrial contractile activity. AB - OBJECTIVE: The objective of this study was to test the hypothesis that cocaine acutely increases contractile activity in isolated pregnant human myometrium. STUDY DESIGN: Myometrial samples were obtained from the lower uterine segment at elective cesarean section from five women at term who were not in labor and who had no perinatal risk factors. Myometrial strips were suspended in contractile buffer, and isometric contractions were measured. Frequency, amplitude, duration, and integrated area (mean +/- SE) were compared before and after the addition of cocaine (10(-6) to 10(-4) mol/L) by means of analysis of variance and Duncan's multiple range test. RESULTS: Contraction duration, expressed relative to control, increased acutely after addition of cocaine (10(-5) mol/L, 2.0 +/- 0.29; 10(-4) mol/L, 2.8 +/- 0.64) (p < 0.001). Integrated area of contractions also increased relative to control (10(-6) mol/L, 1.6 +/- 0.18, p < 0.05; 10(-5) mol/L, 2.4 +/- 0.16 and 10(-4) mol/L, 3.5 +/- 0.23, p < 0.001). These effects were dose dependent. CONCLUSION: Cocaine acutely increases contractile activity in myometrium isolated from pregnant women. PMID- 8238133 TI - T-type and L-type calcium currents in freshly dispersed human uterine smooth muscle cells. AB - OBJECTIVE: Our purpose was to characterize the types of voltage-activated calcium currents that are found in human uterine myocytes and to determine the effects of magnesium and nifedipine on these currents. STUDY DESIGN: Electrophysiologic experiments were performed on freshly isolated human uterine smooth muscle cells by means of the nystatin modification of the whole-cell patch clamp technique. RESULTS: Two types of voltage-activated calcium currents that are similar to the T-type and L-type calcium currents observed in cardiac myocytes were identified in freshly dispersed, pregnant human uterine myocytes. Magnesium at 8 mmol/L reduced uterine myocyte T-type currents by 68% +/- 17% but did not reduce L-type currents. Nifedipine at 10(-6) mol/L blocked the L-type currents but had no effect on T-type currents. CONCLUSION: Freshly isolated human uterine smooth muscle cells exhibit subtypes of calcium currents that are analogous to those found in cardiac myocytes. The uterine myocyte T-type current may be primarily involved with action potential transmission and the L-type current primarily with increasing intracellular free calcium by bulk calcium transport. The differing physiologic effects of magnesium and nifedipine on the calcium current subtypes suggest that for the treatment of preterm labor the primary effect of magnesium therapy is to decrease the frequency of contractions and of nifedipine the strength. PMID- 8238134 TI - The presence of cervical and vaginal fetal fibronectin predicts preterm delivery in an inner-city obstetric population. AB - OBJECTIVE: It has previously been shown that fibronectin bearing a specific oncofetal domain is present at the chorionic-decidual interface and that its release into cervical and vaginal secretions accurately predicts preterm delivery in patients with uterine contractions. This study examines whether serial assessment of cervical and vaginal fetal fibronectin allows for the prediction of preterm delivery in symptom-free patients derived from an inner-city, general obstetric population. STUDY DESIGN: Cervical and vaginal samples were obtained from 429 consenting patients who received routine prenatal care between 24 and 37 weeks' gestation. A sensitive immunoassay was used to quantitate cervical and vaginal fetal fibronectin levels, and clinicians were blinded to fetal fibronectin results. Post hoc receiver operating characteristic curve analysis was used to determine which sample site (cervical or vaginal), fetal fibronectin concentration, and number of consecutive positive samples optimized screening efficacy. Logistic regression was employed to determine whether fetal fibronectin was an independent predictor of preterm delivery. RESULTS: The spontaneous preterm delivery rate was 11% (49/429). Among the 326 patients sampled within 28 days of delivery, receiver operating characteristic curve analysis indicated that the presence of a single cervical fetal fibronectin value > 60 ng/ml between 24 and 36 weeks' gestation predicted preterm delivery with a sensitivity of 73%, a specificity of 72%, and positive and negative predictive values of 25% and 95%, respectively. A vaginal fetal fibronectin value > 50 ng/ml predicted preterm delivery with a sensitivity of 68%, a specificity of 80%, and positive and negative predictive values of 30% and 95%, respectively. Cervical and vaginal fetal fibronectin predicted preterm deliveries resulting from both membrane rupture and preterm labor with intact membranes. A positive fetal fibronectin result preceded preterm delivery by 3.4 (+/- 3.2) weeks. Stepwise logistic regression demonstrated that cervical and vaginal fetal fibronectin levels were independent predictors of preterm delivery with adjusted odds ratios of 8.9 (95% confidence interval 3.6 to 22.1) and 6.0 (95% confidence interval 2.6 to 13.7), respectively. CONCLUSIONS: Among patients undergoing monthly cervical and vaginal sampling between 24 and 36 weeks' gestation, the presence of fetal fibronectin is a sensitive and specific predictor of preterm delivery. PMID- 8238135 TI - Surfactant replacement therapy in utero for prevention of hyaline membrane disease in the preterm baboon. AB - OBJECTIVE: A previous study of intraamniotic administration of surfactant in preterm rabbit fetuses demonstrated that exogenous surfactant can be taken up into the lungs from amniotic fluid in quantities sufficient to alter pulmonary mechanical properties. On the basis of these findings we chose to test the hypothesis that intraamniotic administration of surfactant to the preterm baboon 24 hours before delivery will prevent the development of clinical and pathological aspects of hyaline membrane disease. STUDY DESIGN: Characteristics of hyaline membrane disease in the preterm baboon model include atelectasis, the formation of hyaline membranes in the airways and distal air saccules, overexpansion of distal airways, and disruption of airways by barotrauma associated with neonatal intensive care practices. Nine preterm baboons were treated with either saline solution (n = 4) or surfactant (n = 5) by intraamniotic injection on 136 to 137 days' gestation. One day later fetuses were delivered by cesarean section and maintained for 24 hours with standard neonatal intensive care techniques. RESULTS: All neonates completed the protocol, and surfactant-treated animals had better clinical courses as documented by Pao2/PAO2 (p < 0.05), FIO2 (p < 0.005), and Paco2 (p < 0.05). Significant radiographic differences (p < 0.02) were noted to be the result of surfactant treatment. No differences in ventilator pressures or neonatal cardiovascular parameters were found. Significant differences in pulmonary pathologic conditions (p < 0.01) were also found, but all animals had some degree of pathologic pulmonary changes associated with hyaline membrane disease. CONCLUSION: Therefore a single treatment with surfactant in utero significantly improved the clinical course but did not completely prevent hyaline membrane disease in an established animal model. PMID- 8238136 TI - Interleukin-1 and tumor necrosis factor stimulate arachidonic acid release and phospholipid metabolism in human myometrial cells. AB - OBJECTIVE: Our aim was to evaluate the effects of the cytokines interleukin-1 and tumor necrosis factor on arachidonic acid release in human myometrial cells. STUDY DESIGN: Primary monolayer cultures of human myometrial cells prelabeled with tritiated arachidonic acid were exposed to interleukin-1 or tumor necrosis factor for varying periods and the release of tritiated arachidonic acid and its loss from phospholipids were measured by radiochromatography. To gain some information on the biologic action of interleukin-1 the contractile response to oxytocin was measured in myometrial strips preincubated with this cytokine. Data were statistically evaluated with analysis of variance or Student's test. RESULTS: Both cytokines caused a dose-dependent increase in tritiated arachidonate release that was suppressed by the protein synthesis inhibitor cycloheximide. Tritiated arachidonic acid release was maximal after 24 hours of stimulation with interleukin-1. Both interleukin-1 and tumor necrosis factor stimulated the release of the isotopically labeled fatty acid from phosphatidylcholine. In addition, interleukin-1 also increased the loss of arachidonic acid from phosphatidic acid and significantly potentiated the oxytocin-evoked myometrial contractility. CONCLUSIONS: Both interleukin-1 and tumor necrosis factor enhance arachidonic acid release, probably by inducing the synthesis of phospholipase A2 and possibly other enzymes involved in the metabolism of phospholipids. In turn, arachidonic acid itself may act as a second messenger, synergizing with other uterotonic agents, as well as serving as the precursor for prostaglandins and various other bioactive eicosanoids. PMID- 8238137 TI - Elevated second-trimester human chorionic gonadotropin and subsequent pregnancy induced hypertension. AB - OBJECTIVE: Pregnancy-induced hypertension is believed to be a disorder of the vascular endothelium, possibly focused in the placenta. Markers such as cellular fibronectin appear early in patients in whom pregnancy-induced hypertension develops. We hypothesized that patients with elevated second-trimester levels of human chorionic gonadotropin would have an increased risk for pregnancy-induced hypertension. STUDY DESIGN: We conducted a cohort study, using a database of patients undergoing second-trimester serum screening for Down syndrome and who were delivered at our institution between January 1990 and August 1991. We included all 180 women with human chorionic gonadotropin levels > or = 2.0 multiples of the median. A sequential sample of 369 women with levels < 2.0 multiples of the median served as the referent group. Standard American College of Obstetricians and Gynecologists definitions of pregnancy-induced hypertension and proteinuria were used. RESULTS: Patients with elevated human chorionic gonadotropin levels were at increased risk for pregnancy-induced hypertension (risk ratio 1.7, 95% confidence interval 1.2 to 2.4) and proteinuric pregnancy induced hypertension (risk ratio 5.1, 95% confidence interval 1.6 to 16.2). Adjusting for potential confounding factors did not alter these associations. CONCLUSIONS: Patients with elevated human chorionic gonadotropin appear to be at higher risk of pregnancy-induced hypertension and proteinuric pregnancy-induced hypertension. Second-trimester human chorionic gonadotropin levels should be further investigated as a potential marker for pregnancy-induced hypertension. PMID- 8238138 TI - The effect of intrapartum epidural analgesia on nulliparous labor: a randomized, controlled, prospective trial. AB - OBJECTIVE: Our purpose was to determine the effect of epidural analgesia on nulliparous labor and delivery. STUDY DESIGN: Normal term nulliparous women in early spontaneous labor were randomized to receive either narcotic or epidural analgesia. RESULTS: When compared with the group receiving narcotic analgesia (n = 45), the group receiving epidural analgesia (n = 48) had a significant prolongation in the first and second stages of labor, an increased requirement for oxytocin augmentation, and a significant slowing in the rate of cervical dilatation. Epidural analgesia was associated with a significant increase in malposition (4.4% vs 18.8%, p < 0.05). Cesarean delivery occurred more frequently in the epidural group (2.2% vs 25%, p < 0.05), primarily related to an increase in cesarean section for dystocia (2.2% vs 16.7%, p < 0.05). CONCLUSIONS: In a randomized, controlled, prospective trial epidural analgesia resulted in a significant prolongation in the first and second stages of labor and a significant increase in the frequency of cesarean delivery, primarily related to dystocia. PMID- 8238139 TI - The association between prenatal stress and infant birth weight and gestational age at birth: a prospective investigation. AB - OBJECTIVE: The aim was to test a model of the influence of maternal prenatal psychosocial stress on birth outcomes after controlling for biomedical risk. STUDY DESIGN: In a prospective study a sociodemographically homogeneous sample of 90 women was assessed during the third trimester with standard, reliable questionnaires that measured episodic and chronic stress, strain (response to stress), and pregnancy-related anxiety. Birth outcomes included infant birth weight, gestational age at birth, and intrapartum complications. Parity and biomedical (antepartum) risk was also coded. Bivariate and multivariate analyses were performed after controlling for the effects of biomedical risk factors. RESULTS: Independent of biomedical risk, each unit increase of prenatal life event stress (from a possible sample range of 14.7 units) was associated with a 55.03 gm decrease in infant birth weight and with a significant increase in the likelihood of low birth weight (odds ratio 1.32), and each unit increase of prenatal pregnancy anxiety (from a possible sample range of 5 units) was associated with a 3-day decrease in gestational age at birth. CONCLUSION: Independent of biomedical risk, maternal prenatal stress factors are significantly associated with infant birth weight and with gestational age at birth. PMID- 8238140 TI - The effects of centrally administered adenosine on fetal sheep heart rate accelerations. AB - OBJECTIVE: We examined the effect of administering a long-acting adenosine analog, L-2-N6-(phenylisopropyl) adenosine, into the cerebrospinal fluid of the fourth ventricle on fetal sheep heart rate accelerations. STUDY DESIGN: Pregnant ewes between 123 and 130 days' gestation were anesthetized, and the fetal head was exteriorized. Catheters were placed in the fourth cerebral ventricle through the foramen magnum and in the brachial artery to record fetal heart rate. Studies were performed in unanesthetized fetuses 4 to 7 days after surgery. Accelerations were defined as a 10 beats/min rise in heart rate for at least 5 seconds. RESULTS: The mean number of accelerations before administration of L-2-N6- (phenylisopropyl) adenosine was 3.9 +/- 0.7 (mean +/- SE) per 10-minute epoch, decreasing to 2.0 +/- 0.7 and 1.4 +/- 0.8 after instillation of 0.2 microgram and 0.5 micrograms of L-2-N6-(phenylisopropyl) adenosine, respectively (p < 0.05). Increasing the L-2-N6-(phenylisopropyl) adenosine dose to 10.0 micrograms resulted in loss of heart rate accelerations. Accelerations were not reduced when theophylline, an adenosine receptor blocker, was given before L-2-N6 (phenylisopropyl) adenosine. CONCLUSION: Increasing amounts of a centrally administered adenosine analog progressively decreased the number of fetal sheep heart rate accelerations, most probably by suppression of brainstem sympathetic outflow. PMID- 8238141 TI - The hemodynamic effects of isometric exercise during late normal pregnancy. AB - OBJECTIVE: Our study was designed to evaluate the hemodynamic effects of isometric exercise in late normal pregnancy. STUDY DESIGN: Study subjects were 10 healthy pregnant volunteers with uncomplicated singleton gestations between 25 and 36 weeks. Doppler methods were used to derive cardiac output, total peripheral resistance, and stroke volume before, during, and after a defined protocol of lower extremity isometric exercise. Hemodynamics and blood pressure were evaluated and compared. RESULTS: Mean arterial blood pressure and total peripheral resistance increased during the performance of isometric effort (mean blood pressure +/- SD was 78.9 +/- 7.3 to 97.5 +/- 8.6 mm Hg; total peripheral resistance +/- SD was 924 +/- 148 to 1153 +/- 18.3 dyne.sec.cm-5; p < 0.002 and p < 0.001, respectively). Cardiac output remained unchanged throughout the study period. CONCLUSION: In advanced normal pregnancy isometric exercise increases the mean arterial blood pressure by raising the total peripheral resistance. PMID- 8238142 TI - Reproducibility of the oral glucose tolerance test in pregnant women. AB - OBJECTIVE: The purpose of this study was to evaluate the reproducibility of the 3 hour oral glucose tolerance test during pregnancy and the potential factors associated with nonreproducible results. STUDY DESIGN: Thirty-eight women with a 1-hour glucose level > or 135 mg/dl had a 100 gm oral glucose tolerance test. During the test samples were obtained for glucose, insulin, cortisol, human placental lactogen, and norepinephrine levels. The oral glucose tolerance test was repeated 1 week later under similar metabolic conditions. RESULTS: The intraassay coefficient of variation in glucose from week 1 to week 2 was < 2%. There were no significant differences in the paired fasting 1-, 2-, or 3-hour glucose concentrations (p = 0.51 to 0.96) or the area under the glucose curve (p = 0.43) from week 1 to week 2, although the mean absolute difference in glucose values ranged from 4 (fasting) to 18 (3 hours) mg/dl. Oral glucose tolerance test results were classified as either normal or abnormal from week 1 to week 2; 16 normal/normal, 13 abnormal/abnormal, seven abnormal/normal, and two normal/abnormal. Norepinephrine (p = 0.03) and insulin (p = 0.05) were significantly greater in week 1 but not in week 2 in the abnormal/normal versus normal/normal and abnormal/abnormal groups. There were no significant differences in cortisol or human placental lactogen levels among groups at any time. CONCLUSIONS: The oral glucose tolerance test was not reproducible for diagnosis in 24% (nine of 38) of pregnant women. We speculate that maternal stress (increased norepinephrine) may have been a factor for the abnormal results in week 1 in the abnormal/normal group. PMID- 8238143 TI - Fetal renal pulsed Doppler waveform in prolonged pregnancies. AB - OBJECTIVE: Our purpose was to determine Doppler waveforms of the fetal human renal artery in prolonged pregnancy in the presence or absence of oligohydramnios. STUDY DESIGN: Fifty patients at or after 40 weeks were studied. Ultrasonography was performed to determine the amniotic fluid index by the four quadrants technique. Two groups of patients were obtained on the basis of the amniotic fluid index. Group 1 had an amniotic fluid index > 5 (normal) (n = 33); group 2 had amniotic fluid index < or = 5 (oligohydramnios) (n = 17). Umbilical artery and fetal renal pulsed Doppler waveforms were determined and analyzed. The fetal renal artery systolic-to-diastolic ratio of the two groups was compared. RESULTS: Fetuses with a low amniotic fluid index had a significantly higher ratio. A significant negative correlation coefficient between amniotic fluid index and fetal renal systolic/diastolic ratio was found (r = -0.435 and p < 0.01). CONCLUSIONS: In prolonged pregnancies there is a significant relationship between the amniotic fluid index and the fetal renal systolic/diastolic ratio. In pregnancies associated with oligohydramnios the systolic/diastolic ratio is significantly higher than in those with normal amniotic fluid volume. These data suggest that intrarenal flow in prolonged pregnancies complicated with oligohydramnios is significantly different. PMID- 8238144 TI - The effect of low-dose aspirin on fetal urine output and amniotic fluid volume. AB - OBJECTIVE: Patients given prostaglandin synthetase inhibitors in doses sufficient to inhibit labor are at risk for developing oligohydramnios (possibly related to a reduction in fetal urine output). We sought to ascertain whether the fetuses of women who received 60 mg of aspirin daily had a lower urine output than those whose mothers were given a placebo. STUDY DESIGN: Nulliparous women with singleton gestations in a double-blind preeclampsia prevention trial were randomly selected at 24 weeks' gestation to receive either 60 mg of aspirin daily or a placebo. Urine output was assessed in 59 fetuses (aspirin 32, placebo 27) by serial ultrasonographic measurement of their bladder volume (volume = 4/3 pi r3). Biochemical evidence of aspirin compliance was defined as an 80% reduction in maternal serum thromboxane B2 levels when comparing values obtained at randomization with those at 34 to 36 weeks' gestation. RESULTS: Visual assessment of amniotic fluid volume was similar in both groups. Four-quadrant amniotic fluid indexes also were similar (13.5 cm in aspirin group vs 12.2 cm in placebo group, p = 0.15). Mean fetal urine outputs were similar in the aspirin (57.7 ml/hr) and placebo (55.1 ml/hr) groups (p = 0.71). Moreover, the 23 women with a fourfold thromboxane B2 reduction had a higher mean fetal urine output (63.5 vs 51.8 ml/hr, p = 0.08) than did the remaining 35 patients. This study has a 96% chance (1-beta) of detecting a 50% (30 ml) reduction in fetal urine output. CONCLUSIONS: Daily maternal ingestion of 60 mg of aspirin did not decrease fetal urine output or amniotic fluid volume. PMID- 8238145 TI - Maternal and fetal cardiovascular responses to methamphetamine in the pregnant sheep. AB - OBJECTIVES: Our purpose was to evaluate the maternal and fetal cardiovascular effects of maternally administered methamphetamine and to determine the extent of placental transfer. STUDY DESIGN: Seven near-term pregnant sheep were surgically instrumented to measure maternal and fetal blood pressure and heart rate, cardiac output, uterine flow, and umbilical flow. A single dose of methamphetamine 1 mg/kg was administered as a bolus to study maternal and fetal cardiovascular responses and placental transfer. RESULTS: Maternal administration of methamphetamine caused increases in maternal blood pressure, heart rate, cardiac output, and systemic vascular resistance and decreased uterine blood flow. Peak maternal changes occurred within 10 minutes after methamphetamine administration and were near baseline by 180 minutes. Fetal blood pressure increased and returned slowly to baseline by 2 hours. After an initial decrease fetal heart rate increased above baseline values over the next 2 hours. Umbilical blood flow also decreased initially and then increased slightly. Fetal pH and PO2 tended to decrease. Maternal and fetal methamphetamine levels reached a maximum of 2.9 and 1.9 micrograms/ml, respectively. Rapid and significant placental transfer, delayed excretion into the amniotic fluid, and slow elimination from the maternal and fetal circulation were demonstrated. CONCLUSION: Methamphetamine readily crosses the ovine placenta, producing significant and long-lasting maternal and fetal cardiovascular effects, which may have long-term consequences, especially if administered repetitively. PMID- 8238146 TI - Maternal and fetal cardiovascular effects and placental transfer of the oxytocin antagonist atosiban in late-gestation pregnant sheep. AB - OBJECTIVES: Atosiban is a synthetic oxytocin antagonist that is currently undergoing dose-ranging clinical trials. To date, no data are available on the cardiovascular effects of combined oxytocin and vasopressin blockade during late pregnancy. Our aims were (1) to determine the effects of atosiban infusion on the maternal and fetal cardiovascular system and on uterine blood flow and (2) to determine the maternal pharmacokinetics and the rate of placental transfer of atosiban. STUDY DESIGN: Five chronically catheterized pregnant sheep were treated with a 2-hour infusion of atosiban (300 micrograms.min-1) at 116 to 126 days' gestation. Maternal and fetal blood pressure and heart rate and uterine blood flow were measured before and during the infusion. Maternal and fetal arterial blood samples were obtained throughout the experiment for measurement of plasma atosiban levels and blood gas values. RESULTS: No significant change in maternal cardiovascular parameters or uterine blood flow were observed. Similarly, no changes in fetal blood pressure and arterial blood gases were present during the infusion of the atosiban. Maternal plasma levels of atosiban reached a maximum of 585.2 +/- 82.2 (ng/ml mean +/- SD) at the end of the infusion and decreased biexponentially with a mean t1/2 alpha of 17 minutes and a mean t1/2 beta of 2.2 hours. Fetal plasma levels of atosiban were at or below the detection limit. CONCLUSION: Atosiban does not significantly affect maternal or fetal cardiovascular parameters when it is administered for 2 hours in late-pregnant sheep. Although significant levels were measured in maternal blood, negligible transfer to the fetus occurred. PMID- 8238147 TI - Acute hemodynamic effects induced by chorionic villus sampling: a preliminary investigation. AB - OBJECTIVE: Our purpose was to evaluate the effects of chorionic villus sampling on the fetal heart rate, the fetal umbilical artery pulsatility index, and the maternal arcuate artery resistance index. STUDY DESIGN: M-mode fetal heart rate, pulsed Doppler fetal umbilical artery pulsatility index, and maternal arcuate artery resistance index measurements were obtained before and immediately after chorionic villus sampling in 50 patients and were compared with 50 procedure free, gestational age--matched controls. Comparisons within groups were performed with paired T tests and mean changes between groups with two-sample T tests, and variance comparisons between groups were performed with F tests. RESULTS: The mean changes (SD) in fetal heart rate, umbilical artery pulsatility index, and arcuate artery resistance index in patients undergoing chorionic villus sampling and in controls were not statistically different from zero or from each other. However, the variance around the mean fetal heart rate change was 17.1 times larger in the chorionic villus sampling group than the control group (F 17.1, degrees of freedom 49,49, p < 0.0001), and the variance around the mean pulsatility index change was 2.7 times larger in the chorionic villus sampling group than in the control group (F 2.7, degrees of freedom 49,49, p = 0.0007). There was no significant difference in mean resistance index change or variance around the mean resistance index change between groups. CONCLUSION: Chorionic villus sampling induces significant but unpredictable fluctuations in fetal heart rate and umbilical artery pulsatility index but does not affect maternal arcuate artery resistance index. PMID- 8238148 TI - The effect of low-dose aspirin on umbilical artery Doppler measurements. AB - OBJECTIVE: Our purpose was to determine if the maternal administration of low dose (60 mg) aspirin from 24 weeks' gestation until delivery is associated with significant changes in longitudinal umbilical artery Doppler index values. STUDY DESIGN: Healthy, young nulliparous women with singleton gestations who were enrolled in a double-blind trial of low-dose aspirin for preeclampsia prevention underwent prerandomization and monthly postrandomization continuous-wave umbilical artery Doppler evaluations. The systolic-to-diastolic ratio and the resistance index were determined at each examination. Compliance with the medical regimen was assessed by pill counts and measurement of maternal serum thromboxane B2 levels. RESULTS: The prerandomization Doppler index values were similar between the two randomization groups in the 538 (aspirin 266, placebo 272) patients studied. We found no significant differences with univariate comparisons and multiple linear regression modeling in the postrandomization Doppler index values (p = 0.21 to 0.96). No differences were found when patients were assessed by randomization assignment group (intent to treat) or by biochemical evidence for compliance and absence of crossover with maternal serum thromboxane B2 levels. CONCLUSION: The use of low-dose aspirin from 24 weeks' gestation until delivery does not significantly affect umbilical artery Doppler index values. PMID- 8238149 TI - Contractile effect of endothelin in human placental veins: role of endothelium prostaglandins and thromboxane. AB - OBJECTIVE: The aim was to study the effects of endothelin-1 on human placental veins and the role of cyclooxygenase products as mediators of these effects. STUDY DESIGN: Rings of placental veins with and without endothelium were suspended in organ chambers filled with physiologic salt solution. After a period of stabilization at optimal basal tension, isometric tensions in the control group were recorded at increasing concentrations of endothelin-1 (10(-10) to 10( 7) mol/L). Rings in the experimental groups were treated with either indomethacin (cyclooxygenase inhibitor, 10(-5) mol/L), dazoxiben (thromboxane synthetase inhibitor, 10(-4) mol/L), or SQ29548 (thromboxane receptor antagonist, 10(-6) mol/L) before addition of endothelin-1. To demonstrate the presence of functional thromboxane receptors in the rings, contractile responses to U-46619 (10(-9) to 10(-6) mol/L), a thromboxane A2 analog were measured. The effectiveness of SQ29548 blockade was tested in rings treated with SQ29548 (10(-6) mol/L) before addition of U-46619. The concentration-response curves of the treated and control groups were compared with the Student paired t test. RESULTS: Endothelin-1 in doses of 10(-10) to 10(-7) mol/L caused concentration-dependent contraction of placental veins. Indomethacin significantly reduced the response of veins with endothelium to low endothelin-1 concentrations (10(-9.5) to 10(-9) mol/L), (p < 0.05). However, it had no effect at higher endothelin-1 concentrations or in vessels without endothelium. The presence of functional thromboxane A2 receptors was confirmed by the vasoconstrictor effect of U-46619 and its blockade by treatment with SQ29548. Neither SQ29548 nor the thromboxane A2 synthesis inhibitor dazoxiben significantly influenced the response to endothelin-1. CONCLUSIONS: These results demonstrated that endothelin-1 is a potent vasoconstrictor in the human placental vein. Although functional thromboxane A2 receptors exist in this vessel, endothelin-1's action is independent of thromboxane A2. Prostaglandins may mediate part of the endothelin-1-induced placental vasoconstriction. However, endothelin-1 acts primarily by a direct effect on vascular smooth muscle cells. PMID- 8238150 TI - Twin-to-twin transfusion syndrome: a subset of the twin oligohydramnios polyhydramnios sequence. AB - OBJECTIVE: Our purpose was to assess the adequacy of currently accepted criteria for the diagnosis of twin-to-twin transfusion syndrome. STUDY DESIGN: Between April 1991 and February 1992 nine consecutive women with twin gestations and ultrasonographic findings of marked growth discordance, coexistent oligohydramnios-polyhydramnios, monochorionic placenta, and same sex of the fetus in the second trimester were evaluated. Mean gestational age at diagnosis was 19 weeks 5 days +/- 5 weeks, and mean growth discordance was 34% +/- 9%. Seven women underwent attempted sequential amniocentesis and cordocentesis at 23 weeks 6 days +/- 2 weeks 4 days to obtain amniotic fluid and cord blood from each fetus. Type O negative, leucocyte-poor, washed adult red blood cells were transfused into the small, oligohydramnic twin during cordocentesis; immediately afterward blood from the polyhydramnic twin was tested with the Kleihauer-Betke stain. RESULTS: The combined procedure was successful in six (86%) of the cases attempted. True twin to-twin transfusion was confirmed in only four (44%) of those initially identified by ultrasonographic criteria. Hemoglobin difference > 5 g/dl was present in only one of the four cases, and in this case the recipient was anemic. CONCLUSIONS: Currently accepted criteria are insufficient for the diagnosis of true twin-to-twin transfusion. Ultrasonographic findings of marked growth discordance, coexistent oligohydramnios-polyhydramnios, monochorionic placenta, and same sex identify the twin oligohydramnios-polyhydramnios sequence. PMID- 8238151 TI - Placental pathology in discordant twins. AB - OBJECTIVE: The aim of this study was to evaluate placental abnormalities in relation to birth weight discordance in dichorionic and monochorionic twins. STUDY DESIGN: The maternal charts and placental abnormalities of 147 structurally normal twin pairs with cords labeled at delivery were reviewed. The placental weight belonging to each twin was determined by measuring the length, width, and thickness in each of the two placental disks. Placental weight, chorionicity, infarction, abruptio placentae, decidual vascular abnormality, villous fibrosis and hypovascularity, chronic villitis, and intraplacental thrombi were also assessed. Birth weight was discordant if > or = 20%. The data were analyzed with chi 2 and analysis of variance after log transformation of skewed discordancy values. RESULTS: Of the 147 twin pairs, 99 were dichorionic and 48 monochorionic. Placental weights were known for 91 dichorionic and 40 monochorionic twins. Of the lighter cotwins in dichorionic twin pairs 36.3% (33/91) belonged to the heavier placenta, 49.5% (45/91) belonged to the lighter placenta, and 14.3% (13/91) had an equal share of the placental weight with the heavier sibling (p < 0.05). In 42.4% (42/99) the lighter dichorionic twin had more placental lesions than the heavier twin, in 38.4% (38/99) the same number of lesions were present in both placentas, and in 19.2% (19/99) the heavier twin had more placental lesions. There was linear correlation between percent discordance and number of placental lesions in the lighter twin. In dichorionic twins 18 of the 99 (18.1%) were discordant. In 77.8% (14/18) the lighter twin had more placental lesions than the heavier twin, in 16.7% (3/18) the number of lesions was the same in both, and in 5.6% (1/18) the heavier twin had one more lesion than the lighter twin (p < 0.05). In monochorionic twins, regardless of birth weight discordance, no differences in placental abnormalities were observed. CONCLUSIONS: In dichorionic twins significant birth weight discordance was attributable not to differences in placental weight but to a greater number of placental lesions in the lighter twin than in the heavier twin (p < 0.05). This did not hold true for monochorionic twins. PMID- 8238152 TI - Factors contributing to the increased cesarean birth rate in older parturient women. AB - OBJECTIVE: Our purpose was to determine factors contributing to the increased use of cesarean section in patients > or = 35 years old. STUDY DESIGN: Data were collected prospectively on nulliparous patients in spontaneous labor with term, singleton pregnancies and vertex presentations. Criteria for the diagnosis of labor were standardized: regular, painful uterine contractions at least once every 5 minutes in the presence of either complete cervical effacement or spontaneous rupture of membranes. The labors of women > or = 35 years old (n = 74) were compared with those of women 20 to 29 years old (n = 275). RESULTS: The cesarean section rate was significantly greater for patients > or = 35 years old (21.6% vs 10.2%, odds ratio 2.4, 95% confidence interval 1.2 to 5.1). Mean birth weights were similar in the two groups, but when birth weight was > or = 3600 gm patients > or = 35 years old were more likely to be delivered by cesarean section (36.7% vs 12.2%, odds ratio 4.0, 95% confidence interval 1.4 to 11.9). There were no differences between the two age groups in physician factors that could explain the disparate rates of cesarean delivery. Indeed, of patients delivered vaginally the older parturients received oxytocin for longer duration (6.4 +/- 2.6 vs 5.0 +/- 3.1 hours, p < 0.05) and at higher maximum doses (12.4 +/- 6.1 vs 9.8 +/- 6.2 mU, p < 0.05). After controlling for potentially confounding variables with multiple logistic regression analysis, maternal age (R = 0.125, p < 0.005), birth weight (R = 0.196, p < 0.001), the need for oxytocin (R = 0.210, p < 0.001), and epidural anesthesia (R = 0.195, p < 0.001) were found to be independently associated with the increased rate of cesarean section. CONCLUSION: We could not identify any controllable physician factors affecting the rate of cesarean section in patients > or = 35 years old. The increased oxytocin requirements and the incidence of dystocia with birth weight > or = 3600 gm suggest that maternal and fetal characteristics contribute to the increased frequency of cesarean section in older parturients. PMID- 8238153 TI - Factors that influence route of delivery--active versus traditional labor management. AB - OBJECTIVE: Our purpose was to compare maternal and fetal factors that influence the route of delivery with active management of labor and a traditional labor management protocol. STUDY DESIGN: Data were collected prospectively on 346 consecutive patients receiving active management of labor and 354 patients who were managed traditionally. Within each group demographic and labor characteristics of patients undergoing cesarean section were compared with those of patients having vaginal deliveries by means of the Student t test, chi 2 analysis, and stepwise logistic regression. RESULTS: With both active management of labor and traditional labor management success in achieving vaginal delivery was related to the station of the fetal vertex at admission, the need for oxytocin augmentation of labor, the uterine response to oxytocin, the use of epidural anesthesia, and the development of chorioamnionitis. By means of multiple logistic regression analysis maternal age, height, payor status, and birth weight were also identified as risk factors for cesarean section with traditional labor management but not with active management of labor. CONCLUSIONS: Differences were identified in risk factors for cesarean section between active management and traditional labor management. Active management of labor may diminish or eliminate some patient characteristics as risk factors for cesarean birth. PMID- 8238154 TI - Uterine rupture after previous cesarean delivery: maternal and fetal consequences. AB - OBJECTIVE: The purpose of this study was to identify the risk factors associated with overt, catastrophic uterine rupture and to report maternal and neonatal outcomes. The associated elapsed time window for delivery of an uncompromised neonate was also investigated. STUDY DESIGN: A retrospective study with review of charts and monitor strips was performed. RESULTS: Between Jan. 1, 1983, and June 30, 1992, there were 106 cases of uterine rupture at our institution. Of these, seven charts were incomplete and excluded; of the remainder, 28 patients had complete, 13 patients had partial, and 58 patients had no fetal extrusion into the maternal abdomen. Maternal characteristics or intrapartum events were not predictive of the catastrophic extent of uterine rupture. There was one maternal death. Complete fetal extrusion was associated with a higher incidence of perinatal mortality and morbidity. Significant neonatal morbidity occurred when > or = 18 minutes elapsed between the onset of prolonged deceleration and delivery. CONCLUSION: Neonatal and maternal complications in uterine rupture with complete fetal extrusion were low with prompt intervention. PMID- 8238155 TI - Neonatal morbidity after preterm delivery in the presence of documented lung maturity. AB - OBJECTIVE: Our purpose was to determine the incidence of significant neonatal morbidity in fetuses with documented pulmonary maturity delivered before 37 weeks' gestation. STUDY DESIGN: A retrospective review of 213 pregnancies with documented fetal lung maturity (lecithin/sphingomyelin ratio > or = 2.0 or phosphatidylglycerol present) and delivery before 37 weeks was performed. The incidence of neonatal respiratory distress syndrome, bronchopulmonary dysplasia, grade 3 or 4 intraventricular hemorrhage, necrotizing enterocolitis, patent ductus arteriosus, retinopathy of prematurity, infectious morbidity, hyperbilirubinemia, and admission to the special care nursery was determined for those pregnancies with intact membranes and preterm premature rupture of membranes. RESULTS: Serious neonatal morbidity declined with advancing gestational age and was less common after 32 completed weeks of pregnancy. Although the frequencies of respiratory distress syndrome, grade 3 or 4 intraventricular hemorrhage, and necrotizing enterocolitis were 19.4% (12/62), 8.1% (5/62), and 4.8% (3/62), respectively, at < or = 33 weeks' gestation, one case of respiratory distress syndrome, one case of grade 3 intraventricular hemorrhage, and one case of necrotizing enterocolitis occurred in the 151 neonates born at > or = 34 weeks' gestation. CONCLUSIONS: In spite of fetal lung maturity major neonatal morbidity was observed in our patient population. These data relating neonatal morbidity to gestational age are useful in the critical decision regarding timing of delivery. PMID- 8238156 TI - Ritodrine increases leukotriene B4 concentrations in pregnant sheep. AB - OBJECTIVES: Our goals were (1) to determine if beta-adrenergic receptor stimulation leads to 5-lipoxygenase metabolism of arachidonic acid and (2) to determine if inhibition of prostaglandin synthase alters 5-lipoxygenase metabolism of arachidonic acid during beta-adrenergic stimulation. STUDY DESIGN: We infused saline solution, ritodrine (4 micrograms/kg/min), and a combination of ritodrine (4 micrograms/kg/min) and ketorolac (1.2 microgram/kg/min) into chronically catheterized pregnant sheep (gestational ages 110 to 120 days, term 147 days). With a radioimmunoassay we measured concentrations of leukotriene B4, a 5-lipoxygenase metabolite of arachidonic acid, in uterine venous and arterial plasma at 0, 2, and 4 hours during the infusion. RESULTS: Both uterine venous and arterial leukotriene B4 were increased during ritodrine infusion (mean uterine venous increase at 2 hours 218%, p < 0.05; mean arterial increase at 2 hours 280%, p < 0.05). Concentrations during combined infusion of ritodrine and ketorolac increased significantly but were not different than concentrations observed during ritodrine infusion. CONCLUSION: Infusion of the beta-agonist ritodrine leads to 5-lipoxygenase metabolism of arachidonic acid and increased concentrations of leukotriene B4. The increased concentration in both uterine venous and arterial plasma suggests a systemic source of leukotriene B4 production. Concurrent inhibition of prostaglandin synthase during ritodrine infusion does not change 5-lipoxygenase metabolism in this model. PMID- 8238157 TI - Efficacy and safety of nifedipine versus magnesium sulfate in the management of preterm labor: a randomized study. AB - OBJECTIVE: Our purpose was to establish the efficacy and safety of nifedipine versus magnesium sulfate in arresting preterm labor and the efficacy of nifedipine versus terbutaline in preventing recurrent labor. STUDY DESIGN: Singleton pregnancies at < 34 weeks in preterm labor were randomized to either oral nifedipine or intravenous magnesium sulfate. In case of tocolysis failure ritodrine was added. After labor was arrested, the patients in the nifedipine group were maintained on oral nifedipine, and those in the magnesium sulfate group were treated with oral terbutaline until completing 34 weeks. RESULTS: Of 100 patients 80 were considered eligible, of whom 39 were randomized to the nifedipine group. Both groups were comparable in terms of a number of entry variables, including cervical examination, contraction frequency, and gestational age. Both drugs were equally effective in arresting labor and delaying delivery > 48 hours, 92% versus 93%. Both study groups had a similar incidence of side effects, although four (10%) of magnesium sulfate-treated patients required drug discontinuation because of severe symptoms. Nifedipine was as effective as terbutaline in preventing recurrent labor, 26% versus 24%, and in achieving a gestation > 34 weeks, 62% versus 68%. CONCLUSIONS: Oral nifedipine is as effective as magnesium sulfate and terbutaline in arresting and preventing idiopathic preterm labor. PMID- 8238158 TI - The efficacy of oral terbutaline after intravenous tocolysis. AB - OBJECTIVE: Our purpose was to investigate in a prospective, randomized study the efficacy of oral terbutaline after successful intravenous tocolysis in reducing preterm birth. STUDY DESIGN: Patients between 28 and 35 weeks' gestation with uterine contractions and change in cervical examination were treated with intravenous magnesium sulfate for 12 to 24 hours. After successful tocolysis patients were approached for study participation and randomized to receive either oral terbutaline or no therapy. The dose of terbutaline was individualized to achieve a maternal pulse > 100 beats/min, and terbutaline was continued until 36 completed weeks of gestation. Recurrent preterm labor (contractions with change in cervical examination) for either group was treated with intravenous magnesium sulfate, and subsequent treatment was based on the previous randomization. RESULTS: Fifty-five patients were enrolled (28 terbutaline, 27 no oral tocolytic). No difference was found between groups with respect to time gained (4.0 +/- 2.7 vs 4.6 +/- 3.1 weeks, p = 0.412), gestational age at delivery (35.6 +/- 2.7 vs 36.1 +/- 2.4 weeks, p = 0.562), > or = 37 weeks at delivery (nine vs 13, p = 0.291), recurrent preterm labor (10 vs four, p = 0.104), recurrent uterine contractions alone (five vs eight, p = 0.527), birth weight (2616 +/- 633 gm vs 2645 +/- 599 gm, p = 0.785), special care nursery admissions (eight vs six, p = 0.759), or neonatal respiratory distress syndrome (three vs two, p = 0.965). CONCLUSION: The use of oral terbutaline after successful parenteral tocolysis failed to reduce the rate of preterm birth. PMID- 8238159 TI - Double-blind; placebo-controlled trial of piperacillin prophylaxis in preterm membrane rupture. AB - OBJECTIVE: We attempted to test whether antibiotic therapy prolongs pregnancy in preterm premature rupture of membranes, because preterm premature rupture of membranes is frequently associated with chorionic-decidual infection. STUDY DESIGN: Women with preterm premature rupture of membranes and a singleton gestation at 24 to 34 completed weeks were randomized to receive either piperacillin 3 gm or placebo intravenously every 6 hours for 72 hours and were managed conservatively until spontaneous delivery, chorioamnionitis, or fetal distress. RESULTS: Between January 1987 and January 1992, a total of 75 patients were randomized to receive piperacillin (n = 38) or placebo (n = 37). There were no differences between the piperacillin group and the placebo group in mean gestational age at randomization (30.2 +/- 3 vs 30.3 +/- 2.9 weeks). However, a greater number of patients had pregnancy prolonged beyond 7 days (42.1% vs 10.8% p = 0.005) and the mean latency period was significantly prolonged (11.4 +/- 18.8 vs 6.1 +/- 13.6 days, p = 0.001) in the piperacillin group compared with the control groups. CONCLUSIONS: Use of intravenous piperacillin for 72 hours in preterm premature rupture of membranes significantly prolongs the latency period between membrane rupture and delivery. PMID- 8238160 TI - Randomized trial of prophylactic antibiotic therapy after preterm amnion rupture. AB - OBJECTIVE: Our purpose was to determine whether maternal administration of prophylactic ampicillin or erythromycin after preterm amnion rupture is associated with maternal or neonatal benefits. STUDY DESIGN: Women with singleton pregnancies between 24 and 33 weeks 6 days of gestation were eligible if they had no immediate indication for delivery. After giving informed consent, patients were randomized either to receive ampicillin (erythromycin if penicillin allergic) until delivery or to enter a control group. Women whose cervical cultures were positive for either group B streptococci or Neisseria gonorrhoeae received treatment. Tocolytics and corticosteroids were not used. RESULTS: From January 1990 to February 1992 117 patients (antibiotics 59, control 58) were recruited and analyzed. Prophylactic antibiotics were associated with a longer latent phase (mean 12 vs 7.0 days, p = 0.004) and fewer maternal infectious complications (29% vs 60%, p = 0.001). A higher incidence of neonatal necrotizing enterocolitis was observed in the treatment group (14% vs 3.5%, p = 0.05). Other neonatal complications, including death, were lower in the treatment group, but none attained statistical significance (p = 0.09 to 0.33). CONCLUSION: The use of prophylactic antibiotics in selected patients after preterm amnion rupture appears to have a demonstrable maternal benefit. Large, multicenter trials may demonstrate a significant neonatal benefit or confirm any adverse outcomes. PMID- 8238161 TI - Multifetal reduction of triplets to twins improves perinatal outcome. AB - OBJECTIVE: Our purpose was to compare the perinatal outcome of triplet pregnancies reduced to twins with the outcomes of continuing triplet pregnancies and twin pregnancies. STUDY DESIGN: Pregnancy outcomes of triplet pregnancies reduced to twins delivered between July 1988 and July 1992 were compared with pregnancy outcomes of continuing triplet and twin pregnancies delivered over the same time period. RESULTS: The mean gestational age at delivery for the reduced triplets was 35.6 weeks, compared with 31.2 weeks in the nonreduced triplets (p = 0.002). The perinatal mortality rate was 30 per 1000 births in the reduction group and 210 per 1000 births in the nonreduced triplets (p < 0.0001). There were no statistically significant differences between the reduced and nonreduced twins. CONCLUSION: Multifetal pregnancy reduction of triplets to twins yields an improved perinatal outcome compared with nonreduced triplets and a similar outcome compared with nonreduced twins. PMID- 8238162 TI - Stuck twin syndrome: outcome in thirty-seven consecutive cases. AB - OBJECTIVE: Our purpose was to further evaluate the role of serial amniocentesis in pregnancies complicated by the "stuck twin" syndrome. STUDY DESIGN: A cohort of 37 consecutive cases of stuck twin syndrome was followed up from 1986 through 1992. Evaluations included gestational age at diagnosis and at delivery, mean number of amniocenteses, volume of amniotic fluid withdrawn, placentation, perinatal complications, fetal survival, and neonatal follow-up. RESULTS: Five pregnancies were terminated, five had no intervention, and 27 underwent serial amniocenteses. The mean number of amniocenteses was 3.4 (range 1 to 6), and mean total amniotic fluid volume withdrawn was 5.8 L (range 0.75 to 4.0). In the serial amniocentesis group mean gestational age was 23.1 weeks (range 16 to 30) at diagnosis and 31.5 weeks (range 20 to 38) at delivery. Eighty-two percent had monochorionic placentas, and 36% had marginal or velamentous cord insertions. Infant survival was 39 of 54 (74%) in the serial amniocentesis group compared with four of 10 (40%) in the nonintervention group (relative risk 0.46, 95% confidence interval 0.24 to 0.90). CONCLUSION: Serial amniocentesis was associated with a 54% reduction in fetal and neonatal death in cases of stuck twin syndrome. PMID- 8238163 TI - The use of electrosurgical techniques in the management of premalignant diseases of the vulva, vagina, and cervix: an excisional rather than an ablative approach. AB - In recent years electrosurgical excision techniques have been advocated for the evaluation and treatment of premalignant diseases of the vulva, vagina, and cervix. Technologic advances in electrosurgical generators and in fine wire loops have made these techniques feasible in the outpatient setting, and considerable experience has been accumulated in Europe and the United States. This article discusses the advantages and disadvantages of this "new" technology with emphasis on the available scientific literature. In summary, electrosurgical excision offers the benefit of providing tissue for complete histopathologic evaluation without compromising the established convenience, safety, and efficacy of ablative techniques. PMID- 8238164 TI - A case-control study of endometrial cancer in relation to cigarette smoking, serum estrogen levels, and alcohol use. AB - OBJECTIVES: The purpose of this study was to evaluate the effect of cigarette smoking and alcohol use on the risk of endometrial cancer. The impact of smoking on serum estrone, estradiol, and androstenedione levels also was examined. STUDY DESIGN: This hospital-based case-control study included 168 women with endometrial carcinoma and 334 control women. RESULTS: Women who were current smokers had a lower risk of endometrial cancer than did women who did not smoke (relative rate 0.69; 95% confidence interval 0.40-1.19), whereas women who were exsmokers and women who had never smoked had similar rates (relative rate 0.83; 95% confidence interval 0.46-1.48). There was little overall association between serum estrogen levels and cigarette smoking, although estradiol levels in overweight control subjects were lower among women who were current smokers than among women who had never smoked. Androstenedione levels were slightly higher among women in the control group who smoked than among women in the control group who did not smoke, but the difference was not statistically significant (p = 0.28, two-tailed). Alcohol use was unrelated to endometrial cancer in this study. CONCLUSIONS: The study provides additional support for the hypothesis that smoking is inversely related to endometrial cancer. The inverse smoking association with endometrial cancer may be more directly related to higher serum androstenedione levels than to lower serum estrogen levels except, perhaps, among overweight women. PMID- 8238165 TI - The effect of forceps delivery on cognitive development. AB - OBJECTIVE: We evaluated the influence of forceps delivery on cognitive development of preschool children. STUDY DESIGN: A cohort of 3413 children who were born at the Kaiser Permanente Medical Center in Oakland, California, were selected at age 5 for intelligence quotient testing with the Peabody Picture Vocabulary Test and the Raven Coloured Progressive Matrices. Data were analyzed by means of the Student t test to compare mean intelligence quotient scores of intervention modes of delivery with spontaneous delivery. RESULTS: No significant difference in intelligence quotient was seen in 1192 children delivered by forceps (114 midforceps) compared with 1499 who were delivered spontaneously. CONCLUSION: The association of forceps delivery with adverse cognitive development cannot be supported. PMID- 8238166 TI - Fetal lung maturity is not accelerated in preeclamptic pregnancies. AB - OBJECTIVE: Our purpose was to determine whether there is an increased incidence of pulmonary maturity in premature fetuses of preeclamptic women compared with fetuses of matched controls. STUDY DESIGN: A matched cohort study design was used. One hundred twenty-seven strictly defined preeclamptic women who had undergone amniocentesis for pulmonary maturity assessment were matched for gestational age, race, and infant gender to nonhypertensive women with preterm labor who had undergone the same procedure. Patients with multiple gestation, ruptured membranes, diabetes, or prior glucocorticoid treatment were excluded. Lung maturity was assessed with the Lumadex foam stability index test, the lecithin/sphingomyelin ratio, and the TDx fetal lung maturity assay. In addition, the occurrence of respiratory distress syndrome was assessed in 69 of the 127 matched pairs who were also matched for mode of delivery and amniocentesis-to delivery interval. RESULTS: There was no significant difference in the incidence of an immature result between the preeclamptic and matched control groups (39.4% vs 38.6%). Likewise, women with mild (n = 63) and severe (n = 64) preeclampsia or with small-for-gestational-age (n = 26) and appropriate-for-gestational-age (n = 101) infants, when analyzed separately, showed no differences in comparison with their matched controls. When analyzed by gestational age subgroups, no specific gestational age range was associated with accelerated maturity. The incidence of respiratory distress syndrome in the 69 matched pairs was slightly, but not significantly, higher in the preeclampsia group (relative risk 1.43, 95% confidence interval 0.94 to 2.37). CONCLUSION: This study does not support the contention that fetuses of preeclamptic women exhibit accelerated lung maturation. PMID- 8238167 TI - Interleukin-6 levels in amniotic fluid in normal and abnormal pregnancies: preeclampsia, small-for-gestational-age fetus, and premature labor. AB - OBJECTIVE: Our purpose was to determine if abnormal amniotic fluid interleukin-6 levels correlate with pregnancies complicated by preeclampsia, small-for gestational-age fetus, and premature labor. STUDY DESIGN: A case control study design was used to compare third-trimester amniotic fluid interleukin-6 levels from women with normal pregnancies (n = 25) and pregnancies complicated by preeclampsia (n = 15), small-for-gestational-age perinates (n = 8), and premature labor (n = 29). RESULTS: Amniotic fluid interleukin-6 levels in women with preeclampsia and small-for-gestational-age perinates were less than in controls (p < 0.01). Interleukin-6 levels in women with preterm labor refractory to tocolysis were higher than those in women with preterm labor responsive to tocolysis (p < 0.01). CONCLUSIONS: Interleukin-6 is normally present in amniotic fluid. In the third trimester of pregnancy low levels correlate with small-for gestational-age fetuses and preeclampsia, whereas elevated levels correlate with preterm labor and delivery. PMID- 8238168 TI - Production of prostacyclin and thromboxane A2 in mononuclear cells from preeclamptic women. AB - OBJECTIVE: Although the changes in prostacyclin and thromboxane A2 levels in preeclampsia have been well studied, little work has previously been done on mononuclear cells, and the cause of the changes in unclear. We determined the prostacyclin and thromboxane A2 production in mononuclear cells and investigated the effect of preeclamptic serum on this production. STUDY DESIGN: The production of prostacyclin and thromboxane A2 was measured by enzyme immunoassay in 16 normal pregnant women, nine preeclamptic women with proteinuria, and six preeclamptic women without proteinuria. Sera from these three groups of women were investigated to see whether the sera have any effect on prostaglandin production. RESULTS: The levels of thromboxane A2 in preeclamptic patients were found to be much higher than those of normal pregnancy, whereas the levels of prostacyclin tended to be lower. Such changes lead to a markedly increased ratio of thromboxane A2 to prostacyclin in preeclamptic patients. There was no difference in the levels of prostacyclin and thromboxane A2 between the preeclamptic patients with and without proteinuria. Serum from preeclamptic patients with proteinuria slightly reduced prostacyclin synthesis in normal pregnancy but significantly increased increased thromboxane A2 production, resulting in a ratio of thromboxane A2 to prostacyclin similar to that of preeclampsia. However, serum from preeclamptic patients without proteinuria failed to exert such effects. CONCLUSION: The imbalance between prostacyclin and thromboxane A2 occurs in mononuclear cells from preeclamptic women, and there is a factor(s) in proteinuric-preeclamptic serum to contribute, in part, to these changes. Our findings also suggested that the cause of abnormal prostaglandin production in preeclampsia was complicated and multifactorial. PMID- 8238169 TI - Fetal growth retardation in infants of multiparous and nulliparous women with preeclampsia. AB - OBJECTIVE: Infants of women with preeclampsia are considered at high risk of fetal growth retardation. The purpose of our study was to determine whether the risk differed by parity. STUDY DESIGN: We compared the outcomes of 133 case patients with preeclampsia (101 nulliparous women and 32 multiparous women) and 132 normotensive control patients (52 nulliparous women and 80 multiparous women) who delivered at Northern California Kaiser Permanente hospitals between 1984 and 1985. RESULTS: Women with preeclampsia were more likely than control patients to deliver a small-for-gestational-age infant (adjusted odds ratio 7.0, 95% confidence interval 2.8 to 18.1). After we controlled for smoking status, age, Quetelet index, and race, multiparous women with preeclampsia were at greater risk of having a small-for-gestational-age infant (adjusted odds ratio 29.4, 95% confidence interval 5.2 to 167.5) than were nulliparous women (adjusted odds ratio 4.1, 95% confidence interval 1.2 to 14.1) when compared with normotensive control patients of similar parity. Although multiparous women with preeclampsia had higher mean arterial pressures and somewhat earlier onsets of elevated mean arterial pressure than nulliparous women with preeclampsia, neither of these variables predicted whether the infants would be small for gestational age. CONCLUSION: Multiparous women with preeclampsia are at higher risk of having an infant with fetal growth retardation than are nulliparous women with preeclampsia. PMID- 8238170 TI - Avoiding serious infections associated with abdominal hysterectomy: a meta analysis of antibiotic prophylaxis. AB - OBJECTIVE: Our objective was to determine whether the use of preoperative antibiotics prevents serious infections associated with total abdominal hysterectomy. STUDY DESIGN: We identified 25 randomized controlled trials of antibiotic prophylaxis that used rigorous protocols. We performed meta-analysis and cumulative meta-analyses for all of the trials, and then we performed separate meta-analysis for cefazolin, metronidazole, and tinidazole. RESULTS: Overall, 21.1% (373 of 1768) of the patients who did not receive antibiotic prophylaxis had serious infections after abdominal hysterectomy. Among patients who received any antibiotics, we found that 9.0% (166/1836) had serious postoperative infections; among those who received cefazolin, metronidazole, or tinidazole, 11.4% (70 of 615), 6.3% (17 of 269), and 5.0% (5 of 101), respectively, had serious postoperative morbidity. The differences in the prevalence of infection between women who received prophylaxis and women who did not receive prophylaxis were statistically significant (any antibiotics, p = 0.00001; cefazolin, p = 0.00021; metronidazole, p = 0.015; and tinidazole, p = 0.034). CONCLUSION: Because preoperative antibiotics are highly effective in the prevention of serious infections associated with total abdominal hysterectomy, we believe they should be used routinely. In addition, we believe that the use of controls who receive no treatment is no longer justified in trials of antibiotic prophylaxis for total abdominal hysterectomy. PMID- 8238171 TI - Conservative management and pregnancy outcome in diethylstilbestrol-exposed women with and without gross genital tract abnormalities. AB - OBJECTIVES: Our goal was to determine the effect of conservative management on pregnancy outcome in diethylstilbestrol-exposed women with and without gross structural lesions of the genital tract. STUDY DESIGN: The study included a case series of women prospectively enrolled over a 10-year period. RESULTS: Pregnancy outcome and both antepartum and intrapartum events occurring in a case series of 120 conservatively managed pregnancies in 50 diethylstilbestrol-exposed women were reviewed. Group A (n = 34, 89 pregnancies) consisted of women with gross upper or lower genital tract lesions associated with diethylstilbestrol exposure, whereas group B (n = 16, 31 pregnancies) consisted of women whose lesions were limited to colposcopic findings. Cerclage was limited to women with a history of cervical incompetence (n = 1, two pregnancies) or acute cervical change in the second trimester (one pregnancy). Women with cervical change occurring after 25 weeks' gestation were managed with bed rest. Group A experienced more spontaneous first-trimester losses than group B (25.8% vs 12.9%, p < 0.01), whereas group B had a greater gestational age at delivery (39.8 +/- 1.5 vs 37.3 +/- 3.8 weeks, p < 0.01) than group A. Overall, pregnancies surviving the first trimester (n = 94) resulted in delivery of a viable infant discharged home 92.2% of the time; 77.9% of pregnancies reached term. The perinatal loss rate was 1.3%. Preterm labor (6.5%) and preterm rupture of membranes (3.8%) resulted in an overall rate of preterm birth of 9.2%. There were no significant differences in length of any labor stage between groups, nor were there any differences in the use of oxytocin augmentation or cesarean section rates between groups. Postpartum hemorrhage and retained placenta were infrequent complications. CONCLUSIONS: The majority of pregnancy loss in diethylstilbestrol-exposed patients occurs in the first trimester. Cervical incompetence is an infrequent cause of pregnancy loss even in patients with gross structural abnormalities of the genital tract. Patients who had conservative management had good pregnancy outcomes. Prophylactic cerclage for all diethylstilbestrol-exposed patients should not be recommended. PMID- 8238172 TI - Coitus during pregnancy is not related to bacterial vaginosis or preterm birth. AB - OBJECTIVE: Our goal was to assess the relationship between bacterial vaginosis and sexual intercourse and the impact of both on preterm birth. STUDY DESIGN: The presence of bacterial vaginosis was assessed in 790 healthy nulliparous women between 8 and 17 weeks' gestation, and they were then asked to record weekly the number of occasions of sexual intercourse from registration to term. The end point for each was the occurrence of preterm uterine contractions, preterm birth, or preterm rupture of the membranes. RESULTS: A total of 407 women returned the specific follow-up charts on vaginal intercourse. Bacterial vaginosis was detected at registration in 93 of the 407 subjects (22.9%), but the presence or absence of bacterial vaginosis had no predictive value as regards the frequency of subsequent intercourse. Of the 39 women (9.6%) who experienced threatened preterm labor, 16 were delivered before 37 weeks of gestation. In addition, premature rupture of membranes occurred in 48 women (9 preterm and 39 term). The 95% confidence interval for the weekly frequency of intercourse was calculated for those women delivered at term; a larger (p < 0.05) proportion of women delivered preterm, with (6/9, 66.7%) or without (13/16, 81.3%) premature rupture of membranes, had frequencies less than the 95% confidence interval during the second trimester of pregnancy; the respective proportions were 75.0% (6/8) and 78.6% (11/14) during the third trimester. Furthermore, among those women who had stopped having sexual intercourse in the last trimester, a larger (p < 0.05) proportion was delivered preterm (57.1%), with or without premature rupture of membranes, than was delivered at term (13.6%). In addition, only three women (18.8%) delivered preterm, with (two women) or without (one woman) premature rupture of membranes, reported having intercourse within 1 week before the onset of complications. CONCLUSION: For healthy nulliparous women, coitus during pregnancy is not related to bacterial vaginosis and does not predispose to preterm birth. PMID- 8238173 TI - A comparative study of danazol, a regimen of decreasing doses of danazol, and norethindrone in the treatment of objectively proven unexplained menorrhagia. AB - OBJECTIVE: Our purpose was to compare the efficacy of the recommended dose of danazol, a reduced-dose danazol regimen, and norethindrone in the treatment of objectively proven menorrhagia. Recurrence after discontinuing treatment was also assessed. STUDY DESIGN: The study was a single-blind, randomized, parallel, comparative study. After a placebo run-in period over two menstrual cycles, 57 patients with a baseline mean menstrual blood loss of at least 80 ml per cycle were randomly assigned to receive one of three therapies: danazol, 200 mg/day (n = 19) for three menstrual cycles; danazol, 200 mg/day for one cycle, 100 mg/day for one cycle, and 50 mg/day for one cycle (n = 19); and norethindrone, 5 mg three times daily on days 19 through 26 of the cycle for three consecutive cycles (n = 19). Patients in whom treatment was successful (those experiencing blood loss < 80 ml) were entered in the follow-up phase of the study, receiving placebo for a maximum of four menstrual cycles. RESULTS: The final menstrual blood loss on treatment was significantly less for those patients who received both danazol regimens compared with those who received norethindrone (p = 0.017 for reducing dose danazol vs norethindrone and p = 0.043 for 200 mg of danazol vs norethindrone). Both danazol treatment regimens were significantly more successful in reducing menstrual blood loss to within the normal range than was norethindrone. The reducing-dose danazol regimen was successful in eight of 17 patients (p = 0.027), and 200 mg of danazol was successful in nine of 19 patients (p = 0.029), compared with the two successes of 18 patients treated with norethindrone. Significantly more recipients of 200 mg of danazol than of norethindrone subjectively rated their treatment to be moderately or highly effective (p = 0.033). Both danazol treatment regimens were associated with a higher incidence of adverse events than was norethindrone therapy, although the number of withdrawals were similar and infrequent in the three groups. CONCLUSIONS: Both danazol regimens were significantly more effective than norethindrone in reducing the excessive menstrual blood loss of women with unexplained menorrhagia. A subjective assessment by patients found that only the 200 mg of danazol was judged to be significantly more effective than norethindrone in controlling the heaviness of menstrual bleeding. The reduced dose danazol regimen did not appear to markedly diminish the incidence of adverse events compared with the 200 mg of danazol regimen. PMID- 8238174 TI - The effect of colonization with group B streptococci on the latency phase of patients with preterm premature rupture of membranes. AB - OBJECTIVE: Our purpose was to determine whether colonization of pregnant women with group B streptococci shortens the latency time from rupture of membranes to delivery in patients with preterm premature rupture of membranes. STUDY DESIGN: All patients transferred to Long Beach Memorial Women's Hospital with the diagnosis of preterm premature rupture of the membranes were prospectively recorded. The patients included in the study were transferred between Jan. 1, 1986, and June 30, 1991. Data were collected in regard to various obstetric characteristics, the presence or absence of a digital vaginal examination, tocolytic usage, antibiotic usage, and results of the culture for group B streptococci. The latency period was defined as days from membrane rupture to delivery. Patients with multiple gestations, cerclage in place, advanced labor on admission, or an indicated delivery on admission were excluded from data analysis. Patients with a positive culture for group B streptococci were then compared with those having a negative culture in regard to latency time after membrane rupture. RESULTS: The study population was made up of 332 patients. Forty-three (13%) were positive for group B streptococci; they were compared with 289 that were culture negative. There was no difference in the latency period from membrane rupture to delivery when we controlled for digital vaginal examinations and antibiotic usage. In addition, all other comparisons between the two groups were not significant. CONCLUSION: When the presence of a digital vaginal examination, antibiotic usage, and tocolytic usage are controlled for, colonization with group B streptococci by itself does not appear to affect the latency time from premature rupture of membranes to delivery. PMID- 8238175 TI - Test performance of erythrocyte sedimentation rate and C-reactive protein in assessing the severity of acute pelvic inflammatory disease. AB - OBJECTIVE: The objective of the study was to evaluate the test performance of erythrocyte sedimentation rate and serum C-reactive protein in assessing the severity of acute pelvic inflammatory disease and to determine clinically useful cutoff levels to discriminate mild from severe pelvic inflammatory disease. STUDY DESIGN: The study population consisted of 72 women with acute pelvic inflammatory disease verified by laparoscopy and endometrial histopathologic studies; 37 patients had mild and 35 had severe pelvic inflammatory disease. Cutoff levels for erythrocyte sedimentation rate and C-reactive protein were determined to reach best sensitivity and specificity to discriminate between severe and mild disease. Clinical and microbiologic data were analyzed by chi 2, or t test. Logistic regression analysis was used to analyze risk factors for severe pelvic inflammatory disease. RESULTS: Patients with severe pelvic inflammatory disease had higher erythrocyte sedimentation rates and C-reactive protein levels than did those with mild disease. In detecting severe disease an erythrocyte sedimentation rate > or = 40 mm/hr and C-reactive protein levels > or = 60 mg/L had a sensitivity of 97%, a specificity of 61%, a negative predictive value of 96%, and a positive predictive value of 70%. All patients with tuboovarian abscess or perihepatitis and six of seven patients who had anaerobic bacteria isolated from the fallopian tubes tested positive with these cutoff levels. CONCLUSION: Combined use of erythrocyte sedimentation rate and C-reactive protein levels is useful in assessing the severity of acute pelvic inflammatory disease and augments the clinical decision making regarding treatment. PMID- 8238176 TI - Maternal serum creatine kinase: a possible predictor of tubal pregnancy. AB - Tubal pregnancy remains a clinical challenge in spite of improvements in diagnostic tests and procedures. In a prospective study we evaluated the role of the serum creatine kinase as a possible marker for tubal pregnancy. Three groups of 17 patients each were included in the study: group A, documented tubal pregnancy; group B, spontaneous abortion; and group C, normal pregnancy. Serum was tested for creatine kinase in all of the patients. Creatine kinase level was > 45 IU/L in all patients with tubal pregnancy, significantly higher than the level in patients in either of the two other groups (p < 0.0001). We conclude that a high serum creatine kinase level can be an important diagnostic test in the evaluation of a suspected tubal pregnancy. PMID- 8238177 TI - Plymouth randomized trial of cardiotocogram only versus ST waveform plus cardiotocogram for intrapartum monitoring in 2400 cases. AB - OBJECTIVE: The physiology of changes in the ST waveform of the fetal electrocardiogram has been elucidated in extensive animal and human observational studies. A combination of heart rate and ST waveform analysis might improve the predictive value of intrapartum monitoring. Our purpose was to compare operative intervention and neonatal outcome in labors monitored by the conventional cardiotocogram with those monitored by ST waveform plus the cardiotocogram. STUDY DESIGN: A prospective, randomized clinical trial was performed on 2434 high-risk labors in a district general hospital in Plymouth, England. Statistical analysis was performed by Student t test and chi 2 analysis. RESULTS: There was a 46% reduction (p < 0.001, odds ratio 1.85 [1.35-2.66]) in operative deliveries for "fetal distress" and a trend to less metabolic acidosis (p = 0.09, odds ratio 0.38 [0.13-1.07]) and fewer low 5-minute Apgar scores (p = 0.12, odds ratio 0.62 [0.35-1.08]) in the ST waveform plus cardiotocogram arm. CONCLUSIONS: ST waveform analysis discriminates cardiotocogram changes in labor, and the protocol for interpretation is safe. Further randomized studies are warranted. PMID- 8238178 TI - Endotoxin and interleukin-1 alpha in the cervical mucus and vaginal fluid of pregnant women with bacterial vaginosis. AB - OBJECTIVE: The purpose of our study was to determine the concentrations of endotoxin and interleukin-1 alpha in the cervical mucus and vaginal fluid of pregnant women who either did or did not have bacterial vaginosis. STUDY DESIGN: Samples of cervical mucus and vaginal fluid were collected from women in early pregnancy who had signs of bacterial vaginosis and from healthy control subjects. The samples were analyzed for the concentrations of endotoxin and interleukin-1 alpha. In addition, wet mounts were examined for signs of inflammation indicated by increased numbers of leukocytes. RESULTS: Both endotoxin and interleukin-1 alpha occurred in much higher concentrations (p < 0.0001, p < 0.0002) in both the cervical mucus and the vaginal fluid of women with signs of bacterial vaginosis than they did in healthy control subjects. A correlation was found between the interleukin-1 alpha concentrations in the vaginal fluid and the number of leukocytes as judged by a semi-quantitative evaluation of wet mounts (p = 0.0365). The concentrations of endotoxin correlated with those of interleukin-1 alpha in both fluids (vaginal fluid, p < 0.01; cervical mucus, p < 0.01). CONCLUSION: Our study shows that concentrations of endotoxin and interleukin-1 alpha in cervical mucus and vaginal fluid of women in early pregnancy who have bacterial vaginosis are significantly higher than the corresponding levels in control subjects. PMID- 8238179 TI - Absent umbilical artery diastolic flow in a fetus with a partial mole at 18 weeks' gestation. AB - We present a case in which Doppler velocimetry of the umbilical artery in a fetus with a partial mole at 18 weeks' gestation revealed absent diastolic flow. To the best of our knowledge, this is the first report of Doppler velocimetry of the umbilical artery in this rare condition. Increased placental resistance in this abnormal placenta may explain this finding. PMID- 8238180 TI - Preeclampsia is not associated with altered platelet vasopressin binding and cytosolic Ca++ concentration. AB - OBJECTIVES: Preeclampsia is an important cause of fetal and maternal morbidity and mortality. Recently it was described that platelet cytosolic Ca++ levels could be used to screen for preeclampsia. The current study investigated platelet arginine vasopressin receptor characteristics, platelet cytosolic Ca++ levels, plasma- and platelet-bound arginine vasopressin in white pregnant women. STUDY DESIGN: In a cross-sectional study nine third-trimester nulliparous pregnant women with gestational hypertension (seven with proteinuria, two with excessive weight gain without proteinuria) were compared with nine healthy nulliparous pregnant women matched for gestation length and age and 10 healthy age-matched nonpregnant women. Determined were (1) platelet arginine vasopressin receptor number and affinity, (2) platelet cytosolic Ca++ levels, both basal and on arginine vasopressin or thrombin stimulation, and (3) plasma- and platelet-bound arginine vasopressin levels. RESULTS: None of the measured parameters differed significantly among the three groups studied. Mean arginine vasopressin receptor number and affinity ranged from 108 to 143 receptors per platelet and 0.35 to 0.40 nmol/L, respectively. A single population of binding sites was found (Hill number 0.96). Basal Ca++ levels ranged from 113.4 to 133.3 nmol/L, on arginine vasopressin stimulation from 199 to 250 nmol/L. Median arginine vasopressin levels in platelet-poor plasma were between 1.2 and 2.4 pg/ml, with circulating platelets being estimated to possess two to five molecules of arginine vasopressin per platelet. A significant correlation was found between platelet cytosolic Ca++ levels before and after arginine vasopressin stimulation (r = 0.69, p < 0.001) and a weak correlation between platelet receptor density and arginine vasopressin-stimulated platelet cytosolic Ca++ levels (r = 0.38, p < 0.05). CONCLUSIONS: The studied parameters, platelet cytosolic Ca++ levels, whether basal or after stimulation with arginine vasopressin and vasopressin platelet receptor density and affinity, do not discriminate already hypertensive or preeclamptic white women from nondiseased subjects. A valuable test to screen for preeclampsia awaits further prospective studies. PMID- 8238181 TI - Presence of cytomegalovirus inclusion bodies in a recurrent ulcerative vaginal lesion. AB - Cytomegalovirus is present in the female lower genital tract with an incidence of 4% to 12%. Intracellular inclusion bodies, which constitute evidence of the presence of this organism, are noted on Papanicolaou smears or at direct biopsy. All previous reports regarding tissue diagnosis have involved the uterine cervix. We report an unusual case of a postmenopausal woman who was found to have recurrent symptomatic vaginal lesions; repeated biopsies of the lesion disclosed the presence of cytomegalovirus inclusion bodies. PMID- 8238182 TI - Are pill counts valid measures of compliance in clinical obstetric trials? AB - We compared pill counts with a biochemical measure of compliance in 283 women who participated in a randomized double-blind trial that evaluated the efficacy of low-dose aspirin in the prevention of preeclampsia. Subjects whose pill counts indicated a usage > 100% were less compliant than women with lower pill counts. PMID- 8238183 TI - A comparative metabolic study of two low-estrogen-dose oral contraceptives containing desogestrel or gestodene progestins. AB - OBJECTIVES: Our objective was to compare the effects of low-estrogen-dose oral contraceptives containing desogestrel or gestodene progestins on metabolic risk markers for coronary heart disease. STUDY DESIGN: A cross-sectional comparison of 70 women who used a formulation that contained 30 micrograms ethinyl estradiol and 150 micrograms desogestrel, 43 women who used a formulation that contained 30 micrograms ethinyl estradiol and 75 micrograms gestodene, and 54 women who did not use steroidal contraceptives was performed. RESULTS: Oral contraceptive users had higher concentrations of high-density lipoproteins than did women in the control group (+10% to +20%, p < 0.001) primarily because of increases in high density lipoprotein subfraction 3. High-density lipoprotein subfraction 2 concentrations were higher in users of the desogestrel formulation. Low-density lipoprotein cholesterol concentrations were normal in oral contraceptive users, but triglyceride concentrations were high (+80% to +100%, p < 0.001). Fasting glucose, insulin, and C-peptide concentrations were similar in the three groups, but their responses to a glucose load were higher in oral contraceptive users than in controls (p < 0.01 to p < 0.001). The late plasma insulin response to glucose was higher in the women who used the gestodene formulation than in those who used the desogestrel formulation. CONCLUSIONS: The metabolic profiles induced by these oral contraceptives were remarkably similar and may reflect the activity of the estrogen component. PMID- 8238184 TI - Transvaginal ultrasonographic measurements of the fetal adrenal glands at 12 to 17 weeks of gestation. AB - OBJECTIVE: Our purpose was to compile normative data of fetal adrenal gland measurements between 12 and 17 weeks' gestation with transvaginal ultrasonography. STUDY DESIGN: Transvaginal ultrasonographic measurements of the fetal adrenal length and adrenal-to-kidney length ratio calculations were performed on 100 normal fetuses at 12 to 17 weeks' gestation. The diameters of the fetal adrenals and kidneys on each side were measured, and nomograms were generated. These parameters were also measured in two fetuses whose mothers were treated for congenital adrenal hyperplasia. RESULTS: The measurements of the maximal longitudinal axis of the right and left fetal adrenal glands showed a linear increase with fetal age between 12 and 17 weeks of gestation. The adrenal to-kidney length ratio decreased linearly between these gestational ages. In maternal congenital adrenal hyperplasia treated with glucocorticosteroids the fetal adrenals were small. CONCLUSION: Fetal adrenal gland measurements in the early second trimester show a linear progressive growth between 12 and 17 weeks' gestation. PMID- 8238185 TI - Long-term psychosexual and psychosocial performance of patients with a sigmoid neovagina. AB - OBJECTIVE: We aimed to study the long-term psychosexual and psychosocial performance of patients with a sigmoid vagina in vaginal dysgenesis and male-to female transsexuals. STUDY DESIGN: Nineteen patients who had undergone surgery to create a neovagina with the use of a sigmoid segment participated in a long-term follow-up study. Evaluation was done with standardized gynecologic examination and a structured interview. RESULTS: The anatomic result was good in 18 patients, although several reoperative procedures had been necessary. Sexual adjustment was good or satisfactory in 12 of 19 patients. Of the 19 patients, 16 were capable of reaching an organism. Social adjustment was good or satisfactory in 16 patients. CONCLUSIONS: The creation of a sigmoid neovagina results in a long-term anatomically satisfactory situation, in both patients with vaginal dysgenesis and male-to-female transsexuals. In a majority of the patients sexual and social adjustment is good or satisfactory. PMID- 8238186 TI - Effects of rotation discipline on medical student grades in obstetrics and gynecology throughout the academic year. AB - OBJECTIVES: Our purpose was to determine whether the sequence of rotation disciplines taken can effect medical student examination scores on the National Board of Medical Examiners Subject Exam score for obstetrics and gynecology. STUDY DESIGN: A retrospective study was conducted of 439 student files for the academic years 1987 through 1991. The final clerkship grades and subject examination scores for internal medicine, pediatrics, psychiatry, surgery, and obstetrics and gynecology were reviewed. RESULTS: It was determined that completion of internal medicine or surgery before obstetrics and gynecology yielded higher scores in obstetrics and gynecology (p < 0.00001), with the largest improvement (from 533 to 586) observed when obstetrics and gynecology followed the internal medicine rotation. CONCLUSION: The sequence of rotation can affect scores on the National Board of Medical Examiners subject examination in obstetrics and gynecology. The largest improvement in student scores was observed when obstetrics and gynecology followed the internal medicine or surgery clerkships. PMID- 8238187 TI - Transvaginal ultrasonography in persistent trophoblastic tumor. AB - OBJECTIVE: We assessed the reliability of transvaginal ultrasonography in the detection of uterine involvement in cases of gestational trophoblastic tumor, to establish a possible role of this procedure in the management of such neoplasia. STUDY DESIGN: Transvaginal ultrasonography was performed in six women with gestational trophoblastic tumor at initial diagnosis, during the cytotoxic course when negative beta-human chorionic gonadotrophin levels were obtained, and within 3 to 6 months after the end of chemotherapy. RESULTS: In all cases in which metastatic disease was absent intrauterine localization was easily detected by transvaginal ultrasonography; it appeared as endometrial hypoechoic areas and intramyometrial nodules. Favorable response to chemotherapy was determined by a negative serum beta-human chorionic gonadotropin determination accompanied by the finding of regression of nodules at transvaginal ultrasonography. CONCLUSION: Our data support the introduction of transvaginal ultrasonography in the management of gestational trophoblastic tumor. PMID- 8238188 TI - Excessive estradiol secretion in polycystic ovarian disease. AB - Polycystic ovarian disease is both a hyperestrogenic and a hyperandrogenic syndrome, and all studies have shown that hyperestrogenemia is the result of an elevation of estrone with plasma estradiol levels in the normal follicular range. Because a literature search failed to reveal any report of polycystic ovarian disease with significantly elevated estradiol levels, we report a case in which the plasma estradiol was so massively elevated as to mimic an estrogen-producing neoplasm. This case also suggests that although polycystic ovarian disease is a very rare cause of such excessive estradiol production, it should be included in the differential diagnosis of estrogen-producing neoplasms. PMID- 8238189 TI - Blood flow variations in internal carotid and middle cerebral arteries induced by postmenopausal hormone replacement therapy. AB - OBJECTIVE: Our purpose was to clarify the mechanisms by which postmenopausal estrogen replacement therapy exerts its protective effect on cardiovascular risk. STUDY DESIGN: By means of a bidirectional Doppler ultrasonographic system we measured pulsatility index variations the internal carotid artery and middle cerebral artery in 25 early postmenopausal women during a 6-month period of hormone replacement therapy. Transdermal estradiol (50 micrograms/day) was continuously administered. A 12-day course of medroxyprogesterone acetate (10 mg/day) was added every second month. RESULTS: The pulsatility index showed a significant (p = 0.0001) reduction in both arteries after 6 weeks. At 22 weeks a 25% reduction was measured. No variation of the estrogen-induced pulsatility index reduction was observed at the end of every cyclic progestogen supplementation. CONCLUSIONS: In early postmenopausal women hormone replacement therapy causes a rapid reduction of pulsatility index in brain arteries. Cyclical progestational supplementation does not modify this positive effect on reactivity of the blood vessels. PMID- 8238190 TI - Do survival and morbidity of very-low-birth-weight infants vary according to the primary pregnancy complication that results in preterm delivery? AB - OBJECTIVE: This retrospective study was conducted to determine whether predischarge survival and morbidity of very-low-birth weight infants varied according to the principal pregnancy complication that led to preterm delivery. STUDY DESIGN: The hospital records of 535 consecutive live-born singleton infants who weighed between 500 and 1499 gm were reviewed, and five primary complications that resulted in preterm delivery were identified: (1) premature rupture of membranes (n = 244, 46%), (2) idiopathic preterm labor (n = 97, 18%), (3) antepartum hemorrhage (n = 58, 11%), (4) pregnancy-induced hypertension (n = 98, 18%), and (5) "other" complications (n = 38, 7%). Neonatal records were studied to identify the presence of respiratory distress syndrome, bronchopulmonary dysplasia, pulmonary interstitial emphysema, patent ductus arteriosus, necrotizing enterocolitis, intraventricular hemorrhage, retinopathy of prematurity, and infant death before hospital discharge. Logistic regression analysis was used to analyze the association of each pregnancy complication with the various forms of neonatal morbidity. RESULTS: There were no statistically significant differences in discharge survival rates (range 71% to 88%) among infants born to women who experienced one of the five types of primary complications. Independent of all confounders, premature rupture of membranes was associated with a decreased risk of respiratory distress syndrome, bronchopulmonary dysplasia, pulmonary interstitial emphysema, patent ductus arteriosus, and intraventricular hemorrhage. Preterm labor was associated with an increased risk of pulmonary interstitial emphysema, patent ductus arteriosus, and intraventricular hemorrhage. Pregnancy-induced hypertension was associated with an increased risk of respiratory distress syndrome, pulmonary interstitial emphysema, and patent ductus arteriosus. Antepartum hemorrhage was associated with an increased risk of patent ductus arteriosus. CONCLUSION: The principal pregnancy complication that led to preterm delivery significantly influenced predischarge morbidity but not the predischarge survival of live-born infants. PMID- 8238191 TI - The effect of provider education on blood utilization practices. AB - OBJECTIVE: Our aim was to determine prospectively if a process of provider education and subsequent audit of transfusion criteria can reduce inappropriate blood product infusion. STUDY DESIGN: Beginning in December 1990, a year-long process of provider education and quality assurance audit, with the use of guidelines based on National Institutes of Health blood product consensus conferences, was instituted. After this orientation and education period, the medical records were reviewed for patients on the obstetrics and gynecology service from Dec. 1, 1990, through Sept. 30, 1991, who received blood products (packed red blood cells, fresh-frozen plasma, cryoprecipitate). This group was compared with patients cared for by our service over a similar period before the institution of these guidelines. The incidence of cesarean birth and operative intervention for gynecologic malignancies, as well as the number of major operative gynecologic procedures, was found to be unchanged during the total study period (Dec. 1, 1988, through Sept. 30, 1991). RESULTS: In 1989 there were 1236 units of blood products transfused; in contrast, in 1991 428 units of blood were transfused. In 1989, 107.9 +/- 59.9 units of packed cells per month were used in 33.6 +/- 16.8 patients (3.2 units per patient). In contrast, in 1991, 14.4 +/- 5.8 patients per month received transfusions (2.82 units per patient) with 40.7 +/- 17.2 units of packed cells (p < 0.0001). This represents a 75% decrease in the total number of packed cells and a 60% decrease in the number of patients undergoing transfusion per month. Similar reductions in the usage of cryoprecipitate and fresh-frozen plasma were noted (p = 0.024 and 0.002, respectively). Acute operative blood loss was the most common indication for transfusion. Abdominal hysterectomy was the most common procedure followed by exploratory laparotomy and cesarean section. No patients in whom blood was not used had untoward effects. CONCLUSION: Education as to appropriate blood utilization and concurrent quality assurance audit techniques can safely reduce blood usage on a busy obstetrics and gynecology service in a tertiary care center. PMID- 8238192 TI - Ultrasonographically guided subclavian vein catheterization in critical care obstetrics and gynecologic oncology. AB - Invasive hemodynamic monitoring has become an integral part of intensive care management. Whereas the pulmonary artery catheter is the mainstay for determination of the hemodynamic profile and differentiation between cardiogenic and noncardiogenic forms of pulmonary edema, immediate central venous access in itself is of importance in rapid volume replacement in cases complicated by severe hypovolemia. Central venous catheters are also the preferred route of administration of total parenteral nutrition to patients with cancer. We present our experience with real-time ultrasonographic guidance during subclavian vein catheterization in critical care obstetrics and gynecologic oncology. PMID- 8238193 TI - Sustained-release subdermal estradiol implants: a new alternative in estrogen replacement therapy. AB - OBJECTIVE: This trial was a therapeutic and pharmacokinetic dose-finding study with subdermal implants that showed a constant in vitro daily release rate of estradiol. The aim was to find the daily release rate of estradiol that produces serum estradiol concentrations comparable with those obtained with transdermal estradiol at 0.05 mg/day. STUDY DESIGN: The study was an open crossover comparison of transdermal and subcutaneous delivery systems. Thirty-six postmenopausal (serum follicle-stimulating hormone concentration > 30 IU/L before commencing the study) were treated with transdermal estradiol for 4 weeks. Immediately after this transdermal estradiol delivery was replaced by either one or three estradiol implants. The serum concentrations of estrone, estradiol, follicle-stimulating hormone, and sex hormone-binding globulin were followed up for 8 weeks on implant therapy and compared with those during transdermal estrogen administration. To oppose the estrogen effect on the endometrium, each patient received an intrauterine device releasing a progestin, levonorgestrel. RESULTS: The serum estradiol concentrations achieved with three implants were comparable with those during transdermal administration of estradiol at 0.05 mg/day. Suppression of follicle-stimulating hormone was also better maintained with three implants compared with one. CONCLUSIONS: This study shows that constant release profiles can be achieved with nonbiodegradable estradiol implants. The results suggest that a single application should give sufficient estrogen substitution for more than a year. PMID- 8238194 TI - Nonsteroidal antiinflammatory drugs and reproduction. AB - The role of nonsteroidal antiinflammatory drugs in human reproduction and reproductive disorders is reviewed. Nonsteroidal antiinflammatory drugs inhibit the biosynthesis of prostanoids and can cross the placental barrier into the fetal circulation. Nonsteroidal antiinflammatory drugs are the treatment of choice in primary dysmenorrhea and intrauterine contraceptive device-induced dysmenorrhea and menorrhagia. Nonsteroidal antiinflammatory drugs can be used for effective control of menorrhagia, preterm labor, and polyhydramnios and for prevention of preeclampsia, but larger definitive clinical trials are needed. Prostaglandin inhibition with some nonsteroidal antiinflammatory drugs appear promising in the relief of some premenstrual symptoms and in the prevention of postoperative pelvic adhesion formation, but more studies are needed in women. Whereas implantation and tubal mobility are mediated by local prostaglandins, the potential use of nonsteroidal antiinflammatory drugs to suppress prostaglandin in these reproductive processes for enhancing or reducing fertility warrants further studies. In the last 2 decades nonsteroidal antiinflammatory drugs have been used increasingly in the treatment of some reproductive disorders and are promising for many others. PMID- 8238195 TI - Expression of connexin-43 in human myometrium and leiomyoma. AB - OBJECTIVES: The expression of connexin-43 was examined in human leiomyomas and in autologous myometrium. STUDY DESIGN: Indirect immunofluorescence was used to detect connexin-43 gap junctions in myometrial and leiomyoma tissues and in primary cultures. Immunoblot and Northern analyses were used to examine the levels of connexin-43 protein and cx43 messenger ribonucleic acid in primary cultures. RESULTS: Connexin-43 gap junctions were detected in leiomyoma tissue from nonpregnant premenopausal women but not in autologous myometrial tissue. Connexin-43 gap junctions form spontaneously in primary cultures of myometrium and leiomyoma in the presence of 17 beta-estradiol. Fluorescent dye injection confirmed that the gap junctions were coupled in the primary cells. Connexin-43 protein and messenger ribonucleic acid levels generally were higher in leiomyoma cells compared with those of autologous myometrial cells. Steady-state levels of cx43 messenger ribonucleic acid decreased with addition of medroxyprogesterone acetate to the cultures. CONCLUSIONS: Differences in the levels of cx43 expression in leiomyoma and myometrial cells may reflect differences in responses to steroid hormones. PMID- 8238196 TI - The effects of potassium channel openers on isolated pregnant human myometrium before and after the onset of labor: potential for tocolysis. AB - OBJECTIVE: Our purpose was to investigate the effects and pharmacologic properties of potassium channel openers in isolated pregnant human myometrium. STUDY DESIGN: Biopsy specimens of myometrium obtained from 67 women during pregnancy and labor were used for isometric recording under physiologic conditions. RESULTS: Levcromakalim and pinacidil, two prototype potassium channel openers, are potent inhibitors of spontaneous and induced (0.5 nmol/L oxytocin and 10 mumol/L phenylephrine) contractions in isolated human pregnant myometrium, obtained before and after the onset of labor. The sulfonylurea glibenclamide is an apparent competitive antagonist of this inhibition. No antagonism was observed with the sulfonylurea tolbutamide. Both potassium channel openers significantly inhibited contractility evoked by low (10 and 20 mmol/L) but not high (40 and 80 mmol/L) concentrations of extracellular potassium chloride. CONCLUSION: These findings suggest that the relaxant ability of levcromakalim and pinacidil in human pregnant myometrium is because of potassium channel activation. This introduces a potential new approach for tocolysis. PMID- 8238197 TI - Genital and nongenital teratogenesis of prenatal progestogen therapy: the effects of 17 alpha-hydroxyprogesterone caproate on embryonic and fetal development and endochondral ossification in the C57B1/6J mouse. AB - OBJECTIVE: The current study assesses the effects of 17 alpha-hydroxyprogesterone caproate on embryo-fetal development, with a particular focus placed on intrauterine endochondral bone development and growth. STUDY DESIGN: Primigravid C57 BI/6J mice were administered 17 alpha-hydroxyprogesterone caproate by means of subdermal pellets designed to deliver doses of 0.5, 5.0, and 50.0 mg/kg/day on gestational days 7 through 19. Assuming a dosage regimen of 250 mg per 50 kg per week in humans for the treatment of threatened spontaneous abortion, the doses used were 0.7, 7.0, and 70.0 times the human dose equivalent. The time course of exposure encompassed the period of organogenesis through the late fetal period. RESULTS: No maternal toxicity was noted throughout the course of 17 alpha hydroxyprogesterone caproate administration. Fetal weight at term and the percentage of resorptions, dead fetuses, male fetuses, and malformations at term were not significantly affected at any evaluated dose when compared with control group values. The steroid did not exert a significant influence on any assessed parameter of endochondral bone development or growth, even at a dosage concentration that far exceeded the human therapeutic dose equivalent. As would be expected, fetal weight was directly correlated with diaphyseal length and also influenced the relationship of cartilage to osseous tissue in both the humeri and femora. There were no 17 alpha-hydroxyprogesterone caproate dose and fetal sexual phenotype interactive effects noted with regard to any developmental or bone growth parameter measured, thus discounting any sexual phenotype-related toxic or teratogenic sensitivities. CONCLUSION: The data indicate that 17 alpha hydroxyprogesterone caproate administered at doses of up to 70 times the human dose equivalent did not induce increases in the frequency of genital and, importantly, nongenital teratogenesis in spite of exposure during the critical period of organogenesis. Additionally, the steroid did not affect limb morphogenesis or endochondral ossification. PMID- 8238198 TI - Neutrophil attractant/activating peptide-1/interleukin-8: association with histologic chorioamnionitis, preterm delivery, and bioactive amniotic fluid leukoattractants. AB - OBJECTIVES: The goals of this study were (1) to determine immunoreactive neutrophil attractant/activating peptide-1/interleukin-8 levels in amniotic fluid from patients with preterm labor and (2) to compare neutrophil attractant/activating peptide-1/interleukin-8 levels, amniotic fluid culture, Gram stain, and the leukotaxis bioassay for their ability to predict histologic chorioamnionitis and clinical outcome. STUDY DESIGN: Amniotic fluid was collected by amniocentesis from 55 patients with idiopathic preterm labor and three patients with preterm labor and clinical chorioamnionitis. Gram stain, culture (aerobic, anaerobic, and Mycoplasma species), leukotaxis bioassay, and a commercially available neutrophil attractant/activating peptide-1/interleukin-8 enzyme-linked immunosorbent assay (sensitivity 1 ng/ml) were performed on the amniotic fluid samples. Placentas and chorionic membranes were evaluated for evidence of histologic chorioamnionitis in patients delivered preterm. RESULTS: All patients with detectable leukoattractants by the leukotaxis bioassay had neutrophil attractant/activating peptide-1/interleukin-8 levels above the threshold of the assay. The presence of amniotic fluid neutrophil attractant/activating peptide-1/interleukin-8 is a more sensitive marker for histologic chorioamnionitis and delivery before 34 weeks than is amniotic fluid culture (100% vs 59%, p < 0.01; and 95% vs 56%, p < 0.01, respectively). Also, of patients in idiopathic preterm labor those without amniotic fluid leukoattractants (group 1) had the lowest amniotic fluid levels, followed by patients with amniotic fluid leukoattractants and a negative culture (group 2) and patients with amniotic fluid leukoattractants and a positive culture (group 3) who had the highest levels (group 1 vs group 2, p < 0.001; group 2 vs group 3, p < 0.01). CONCLUSIONS: Amniotic fluid neutrophil attractant/activating peptide 1/interleukin-8, like the leukotaxis assay, is an accurate antepartum predictor of histologic chorioamnionitis and subsequent early delivery in patients with preterm onset of labor. This study supports the role of neutrophil attractant/activating peptide-1/interleukin-8 in the recruitment of neutrophils into chorionic membranes and placenta during developing intrauterine infection. PMID- 8238199 TI - Three-dimensional intraabdominal sound pressures in sheep produced by airborne stimuli. AB - OBJECTIVE: Our aims were to investigate how airborne sound was distributed within the abdominal cavity of sheep as function of frequency. STUDY DESIGN: Airborne broad-band noise was measured with a hydrophone at 45 locations within the abdomen of five nonpregnant sheep post mortem and with a microphone extraabdominally. Sound pressure attenuation provided by the abdomen and its contents was determined for frequencies between 50 and 5000 Hz. An analysis of variance was applied to assess the effects of frequency, hydrophone location, and animal on intraabdominal distribution of sound pressures. RESULTS: Below 250 Hz sound pressure was higher inside the animal than outside. Little attenuation (< 10 dB) was found for sounds > 3000 Hz. Attenuation was greatest in the center of the abdomen and least along the inner margin of the abdomen. Intraabdominal sound pressure level varied with frequency (p < 0.0001) and with position of the hydrophone in the cross-sectional plane (p < 0.005) but not in the sagittal plane (p = 0.51). There was no animal effect (p = 0.18). CONCLUSION: During maternal exposure to airborne, broadband noise the fetus could be subjected to intense sound pressures at low frequencies regardless of position within the uterus and at high frequencies when positioned near the abdominal surface. PMID- 8238200 TI - Inhibition of nitric oxide synthesis in rats during pregnancy produces signs similar to those of preeclampsia. AB - OBJECTIVES: Preeclampsia is associated with hypertension, fetal growth retardation, and proteinuria. We hypothesized that impaired vascular nitric oxide synthesis during pregnancy may be an important causal factor in preeclampsia. STUDY DESIGN: An inhibitor of nitric oxide synthase, L-nitro-arginine methyl ester, or a nitric oxide donor, nitroglycerin, was infused subcutaneously to rats at a constant rate from day 17 of gestation. Systolic blood pressure, day of spontaneous delivery, weight, and mortality rate of pups were recorded. RESULTS: Systolic blood pressures in rats infused with L-nitro-arginine methyl ester at daily doses of both 25 and 50 mg were significantly elevated compared with controls. This treatment also caused a substantial decrease in the weight of pups, with an increase in mortality rate, without affecting the gestational length. These effects were dose dependent. Nitroglycerin infusion, on the other hand, affected neither the weight and mortality rate of the pups nor the length of gestation. CONCLUSIONS: Infusion of an inhibitor of nitric oxide synthesis during pregnancy causes hypertension and fetal growth retardation, without affecting gestational length. These signs are similar to those of preeclampsia and indicate that an alteration in nitric oxide synthesis may be one of the factors responsible for this disorder. Treatment with nitric oxide inhibitors may be used in an animal model for preeclampsia, to test various therapeutic strategies. PMID- 8238201 TI - Smoking and preterm labor: effect of a cigarette smoke extract on the secretion of platelet-activating factor-acetylhydrolase by human decidual macrophages. AB - OBJECTIVE: Maternal smoking in pregnancy is associated with a significant increase in the incidence of preterm labor, premature rupture of membranes, and premature delivery. Our aim was to clarify the cause underlying this association. STUDY DESIGN: The effect of cigarette smoke extract on the secretion of platelet activating factor-acetylhydrolase by both decidual macrophages and peripheral blood monocytes and macrophages was investigated. RESULTS: The cigarette smoke extract inhibited the platelet-activating factor-acetylhydrolase secretion by these cells. The inhibitory effect of cigarette smoke extract on the secretion was a hundred times more potent compared with its direct effect on the plasma enzyme. Glutathione and dithiothreitol blocked the inhibition, whereas catalase or superoxide dismutase did not. Nicotine and cotinine have no effect on the secretion. CONCLUSION: The presence in cigarette smoke extract of a potent inhibitor(s) of platelet-activating factor-acetylhydrolase secretion by decidual macrophages may provide an insight into the pathogenesis of preterm labor, premature rupture of membranes, and premature delivery in women who smoke during pregnancy. PMID- 8238202 TI - Gestational changes in L-arginine-induced relaxation of pregnant rat and human myometrial smooth muscle. AB - OBJECTIVE: We intended to demonstrate the presence of an L-arginine-nitric oxide system in human myometrium and to clarify the mechanisms of action of nitric oxide on rat myometrium during gestation. STUDY DESIGN: By examining very small myometrial muscle strips (approximately 750 muscle cells), characteristic features of contraction of rat longitudinal muscle at the midstage of gestation (day 16) and during delivery at term were determined. RESULTS: Spontaneous contractions were significantly different during delivery compared with the midstage of gestation of rat myometrium. L-Arginine relaxed spontaneous and carbachol-induced, but not potassium chloride-evoked, contractions at both stages. However, much higher concentrations of L-arginine were required during delivery, 8-Bromo-cyclic guanosine monophosphate inhibited spontaneous contractions from concentrations of 1 nmol/L in the midstage of gestation and from 0.1 mmol/L during delivery. In human myometrial tissues L-arginine also inhibited contractions during the late stages of gestation. CONCLUSION: (1) The experimental model is sufficient to compare properties of longitudinal myometrial strips during gestation. (2) In rat and human myometrium an L-arginine-nitric oxide system has an important role in inhibiting uterine contractility and possibly maintaining pregnancy. (3) The relaxing effect of the nitric oxide system is largely because of the voltage-independent action of cyclic guanosine monophosphate systems. PMID- 8238203 TI - Pregnancy and lipid peroxide-induced alterations of eicosanoid-metabolizing enzymes in the aorta of the rat. AB - OBJECTIVES: We examined whether pregnancy modulates the expression of prostaglandin endoperoxide synthase, prostacyclin synthase, and thromboxane A2 synthase in the systemic vasculature. Further, we examined whether elevated lipid peroxidation during pregnancy (induced by vitamin E deprivation) affects the normal adaptive process to pregnancy. STUDY DESIGN: Western immunoblotting was performed on aortas from normal and vitamin E-deprived late pregnant (18 to 19 days) and age-matched virgin control rats. RESULTS: Normal pregnancy resulted in an increased expression of prostaglandin endoperoxide synthase (2.91 vs 1.06 fmol/ng deoxyribonucleic acid, p < 0.05). Surprisingly, the expression for both prostacyclin and thromboxane A2 synthases were significantly decreased by pregnancy: prostacyclin synthase 2.60 versus 13.82 units/ng deoxyribonucleic acid and thromboxane A2 synthase 1.32 versus 9.85 units/ng of deoxyribonucleic acid. Elevation of endogenous lipid peroxidation partially reversed this normal pregnancy trend in enzyme expression: prostaglandin endoperoxide synthase 1.85 fmol/ng deoxyribonucleic acid, prostacyclin synthase 9.38 units/ng deoxyribonucleic acid, thromboxane A2 synthase 4.36 units/ng deoxyribonucleic acid. CONCLUSION: Changes in prostanoid activity in the systemic vasculature during pregnancy may be mediated by concerted induction and down-regulation of specific enzymes. Increased lipid peroxidation interferes with this normal pregnant pattern. Further studies on the cell-specific expression of these genes will help to define the cardiovascular role of prostaglandins in pregnancy and in preeclampsia. PMID- 8238204 TI - Developmental changes in atrial natriuretic factor content and localization of its messenger ribonucleic acid in ovine fetal heart. AB - OBJECTIVES: The current study investigated the developmental changes in atrial natriuretic factor peptide content and messenger ribonucleic acid localization in the atria and ventricles of the ovine fetus throughout the second half of gestation. STUDY DESIGN: Ovine fetuses from 67 to 146 days' gestation (term 147 days) and newborn lambs were used for the study. Tissue atrial natriuretic factor contents were determined by radioimmunoassay, and atrial natriuretic factor messenger ribonucleic acid distribution was determined by in situ hybridization. RESULTS: In fetal atria, atrial natriuretic factor peptide levels were much greater than those in the ventricles. The levels in the atria increased with advancing gestation from 70 to 140 days, reflecting an increase in weight of the atrial chambers. A similar trend was not observed in the ventricles. In the atria, atrial natriuretic factor peptide content (per unit protein) reached high levels at 100 to 110 days' gestation; this was associated with an increase in level of atrial natriuretic factor gene expression. In the ventricles, atrial natriuretic factor peptide content and gene expression were very low throughout the second half of gestation, except for a peak in content that occurred at 100 days. Atrial natriuretic factor messenger ribonucleic acid abundance was much greater in the atria than in the ventricles in fetuses from 90 to 130 days' gestation. The distribution of atrial natriuretic factor messenger ribonucleic acid was homogeneous throughout the thickness of the atria and ventricles of the fetal heart. CONCLUSION: During the second half of gestation in the ovine fetus, the expression of atrial natriuretic factor messenger ribonucleic acid in the atria and ventricles paralleled the appearance of the peptide. PMID- 8238205 TI - High intraabdominal pressure: effects on clinical parameters and lung pathology in baboons (Papio cynocephalus and Papio anubis). AB - OBJECTIVE: The risks of a high intraabdominal pressure during laparoscopy are poorly documented. These aspects were studied in a relevant nonhuman primate model, the baboon (Papio anubis, Papio cynocephalus). STUDY DESIGN: The animals used were to be killed at the completion of a vaccine project. They were anesthetized and intubated but allowed to breathe spontaneously. In group 1 (n = 10) a fixed intraabdominal pressure (between 10 and 35 cm H2O [7 and 26 mm Hg]) was applied over 30 minutes. In group 2 (n = 7) the intraabdominal pressure was increased up to a maximum of 105 cm H2O (77 mm Hg). Blood vessels of the parietal peritoneum and omentum were systematically opened from 30 cm H2O (22 mm Hg) onward. Respiration and pulse rate were measured regularly, and the degree of respiratory distress was assessed. After death of the baboons a necropsy was carried out immediately to study the degree of lung atelectasis. RESULTS: In group 1 the respiration rate increased and the heart rate decreased. At an intraabdominal pressure of > or = 30 cm H2O (22 mm Hg) moderate respiratory distress was seen after 20 minutes. In group 2 one animal had a bilateral pneumothorax at a pressure of 60 cm H2O (44 mm Hg) and died at 90 cm H2O (66 mm Hg). Severe respiratory distress was observed in the other six animals. Severe lung atelectasis was present in only one of the five primates with moderate respiratory distress of group 1 and in all baboons of group 2. CONCLUSION: An increase of intraabdominal pressure up to 105 cm H2O (77 mm Hg) combined with intraabdominal blood vessel injury is not lethal in most baboons. PMID- 8238206 TI - Hepatic histopathologic characteristics in HELLP syndrome (hemolysis, elevated liver enzymes, and low platelet count) PMID- 8238207 TI - Clarification needed to cystic fibrosis model. PMID- 8238208 TI - Elevated maternal serum human chorionic gonadotropin increases the chance of adverse pregnancy outcome. PMID- 8238209 TI - Does tertiary center perinatal care lower mortality in infants < 1500 gm? More data needed. PMID- 8238210 TI - Primary ovarian pregnancy successfully treated with methotrexate. PMID- 8238211 TI - Outcomes of very-low-birth-weight infants after maternal corticosteroid therapy before 28 weeks' gestation. PMID- 8238212 TI - Results of transantral orbital decompression in 428 patients with severe Graves' ophthalmopathy. AB - We reviewed records from 428 consecutive patients with severe Graves' ophthalmopathy to determine early and late results after transantral orbital decompression. Optic neuropathy was present in 217 (50.7%) patients. Post operatively, 402 (89%) of 453 eyes with preoperative visual acuity worse than 20/20 improved or remained the same. Visual field scotomas improved or resolved in 245 (91%) of 269 eyes tested pre- and postoperatively. Preoperative papilledema resolved or improved in 99 (94%) of 105 eyes, and preoperative exposure keratitis improved or resolved in 178 (92%) of 195 eyes. Average proptosis reduction was 4.7 mm. Postoperatively, new diplopia developed in 74 (64%) of 116 patients who had no diplopia before orbital decompression, although 300 patients ultimately had strabismus surgery. At late follow-up (N = 293 patients), 226 (77%) had single vision and 44 (15%) had correction with prism. Complications included sinusitis (18 patients), lower eyelid entropion (38 patients), numb lip (23 patients), cerebrospinal fluid leaks (15 patients), and one frontal lobe hematoma (one patient). The average duration of follow-up was 8.7 years. Transantral orbital decompression effectively reduces proptosis and usually corrects optic neuropathy. In other circumstances, the benefits achieved and the side effects incurred must be carefully balanced for each patient before transantral orbital decompression is considered. PMID- 8238213 TI - Measurement and relationship of subarachnoid pressure of the optic nerve to intracranial pressures in fresh cadavers. AB - We performed a study on six fresh cadavers to measure the subarachnoid pressure of the optic nerve and to determine its relationship to intracranial pressure. A lateral orbitotomy via a coronal flap was used to expose the optic nerve. Various intracranial pressures were achieved by saline infusion through a ventriculostomy. The subarachnoid pressure of the optic nerve was measured at 5 and at 25 mm posterior to the globe under conditions of the created intracranial pressure. The effectiveness of optic nerve sheath decompression performed by two standardized surgical techniques was determined experimentally and by theoretic calculations; the role of the bulbous portion of the optic nerve was assessed experimentally. Measurements of the subarachnoid pressure of the optic nerve showed interindividual variation and a linear relationship with the intracranial pressure. Surgical decompression through a 3 x 5-mm window was more effective than were three linear 5-mm incisions. The bulbous portion of the optic nerve was found to have an important role in cerebrospinal fluid circulation. PMID- 8238214 TI - Relaxing retinotomy with silicone oil or long-acting gas in eyes with severe proliferative vitreoretinopathy. Silicone Study Report 5. The Silicone Study Group. AB - In the Silicone Study, 117 of 404 eyes (29%) with severe proliferative vitreoretinopathy (> or = C-3, full-thickness retinal folds in three or more quadrants) enrolled in the study were treated with vitrectomy, underwent a relaxing retinotomy, and were randomly assigned to treatment with long-acting gas or silicone oil. Forty-six eyes (20%) had undergone no previous vitrectomy (group 1); 71 eyes (42%) had undergone previous vitrectomy (group 2) with intraocular gas tamponade (P < .001). Group 1 eyes not undergoing retinotomy had better anatomic (six months) and visual (six and 24 months) outcomes and less hypotony (six months) than eyes that did regardless of tamponade (P < .05). For eyes undergoing retinotomy, silicone oil decreased the likelihood of hypotony (six months, P < .05). These differences were not found in group 2 eyes. We conclude that eyes undergoing a vitreous operation for the first time for the treatment of proliferative vitreoretinopathy can in most instances be successfully treated by conventional techniques without the need for relaxing retinotomy. Retinotomy may be required more often in patients undergoing repeat vitreous surgery for proliferative vitreoretinopathy, in which case both silicone oil and long-acting perflouropropane gas appear to be equally effective. PMID- 8238215 TI - Experimental long-term vitreous replacement with purified and nonpurified perfluorodecalin. AB - Highly purified perfluorodecalin and non-purified perfluorodecalin were injected into 34 rabbit eyes after mechanical vitrectomy, to evaluate the retinal tolerance in long-term vitreous replacement and the effect of purification on the retina. Unpurified perfluorodecalin caused severe inflammation and retinal damage and eight of ten eyes had developed retinal detachment after four weeks. In the eyes injected with highly purified perfluorodecalin, we observed retinal changes of the rod and cone, outer nuclear, and ganglion-cell layers after two weeks, which progressed to a thinning of all layers of the retina by the fourth week and to localized areas of retinal atrophy by the eighth week in the lower part of the eyes. Retinal damage in the upper part occurred after four weeks. At the perfluorocarbon-aqueous interface we observed a band of retinal necrosis that was noticed in all eyes four weeks postoperatively regardless of the substance used. PMID- 8238216 TI - Long-term effects of ceftriaxone treatment on intraocular Lyme borreliosis. AB - Twelve patients with intraocular Lyme borreliosis suffering from vitreitis were treated with ceftriaxone intravenously. Eight patients had definitive Lyme borreliosis, and tentative diagnosis was made in four patients. After treatment with ceftriaxone, all 12 patients were followed up for more than one year. Vitreitis diminished in all patients and visual acuity improved in six patients. The best results were achieved in patients with definitive Lyme borreliosis. PMID- 8238217 TI - Classification of aphakic cystoid macular edema with focal macular electroretinograms. AB - We compared the amplitude and implicit times of the a-waves, b-waves, and oscillatory potentials of the focal macular electroretinograms of 30 eyes with aphakic cystoid macular edema and the healthy fellow eyes. Ten affected eyes were characterized by reduced amplitudes of the oscillatory potentials with normal a wave and b-wave responses (type 1). Nine affected eyes had both reduced amplitudes of the oscillatory potentials and the b-waves (type 2). Ten affected eyes were characterized by reduced amplitude of the oscillatory potentials, the a waves, and the b-waves (type 3). One eye could not be classified. Visual acuities were as follows: type 1, 0.55 (20/36.4); type 2, 0.31 (20/64.5); and type 3, 0.12 (20/166.7). The mean time between cataract surgery and the electroretinographic testing was significantly longer for type 2 and 3 eyes than for type 1 eyes. The differences in the electroretinographic responses between the affected eye and the normal fellow eye suggested either an increased severity or the stage of the cystoid macular edema. PMID- 8238218 TI - Endophthalmitis caused by Staphylococcus aureus. AB - The medical records of 27 patients (27 eyes) with culture-positive Staphylococcus aureus endophthalmitis, diagnosed between December 1984 and February 1992, were reviewed. In 20 of the 27 eyes, endophthalmitis occurred within seven days of ocular surgery. All tested isolates were sensitive to vancomycin, gentamicin, and ceftazidime. In 13 eyes treated by initial vitrectomy and intraocular antibiotic injection, ten eyes achieved 20/400 or better visual acuity, compared to seven of 14 eyes in the initial vitreous tap and intraocular antibiotic injection group. For the eight eyes treated initially with intraocular corticosteroids, seven eyes achieved 20/400 or better visual acuity, compared to ten of 19 eyes in the group receiving no initial intraocular corticosteroids (P = .190). The use of vitrectomy and intraocular corticosteroids with intraocular antibiotics was associated with the best visual outcomes in this group of eyes with endophthalmitis caused by S. aureus. PMID- 8238219 TI - Aqueous humor dynamics in low-tension glaucoma. AB - We measured the diurnal aqueous humor flow in patients with low-tension glaucoma by clearance of topically applied fluorescein. Ten patients with low-tension glaucoma participated in the study, and ten age-matched healthy volunteers served as control subjects. The diurnal aqueous humor flow was 2.58 +/- 0.78 microliters/min (mean +/- standard deviation) in the control group and 2.48 +/- 0.61 microliters/min in the low-tension glaucoma group. The corresponding nocturnal data were 0.98 +/- 0.22 microliters/min and 1.24 +/- 0.45 microliters/min. The differences were not statistically significant. Tonographic facility of outflow and intraocular pressure were also measured. There were no statistically significant differences between the two groups in either facility or pressure. No evidence was found in this small group of patients to support the hypothesis that the normal tension in this disease results from a combination of low facility and low flow. PMID- 8238220 TI - Trabeculectomy, phacoemulsification, and posterior chamber lens implantation with and without 5-fluorouracil. AB - Subconjunctival 5-fluorouracil has been an effective adjunct in glaucoma filtering surgery. We investigated the effectiveness of 5-fluorouracil in primary trabeculectomy combined with phacoemulsification and posterior chamber intraocular lens implantation (glaucoma triple procedure). Seventy-four patients were enrolled and randomly assigned into two groups to receive either no 5 fluorouracil (control patients) or low-dose 5-fluorouracil (mean total dose, 24.8 mg; mean number of injections, 5.0 +/- 1.3). The mean preoperative intraocular pressure was 20.1 +/- 5.4 mm Hg for the 5-fluorouracil group and 21.0 +/- 5.2 mm Hg for the control group (P = .48, Student's unpaired t-test). The mean number of medications was 2.2 +/- 1.0 and 2.0 +/- 1.0 (P = .49, Mann Whitney U test), respectively. At all postoperative visits, there were no statistically significant differences in mean intraocular pressures between the two groups. Mean follow-up was 13.2 months for 5-fluorouracil-treated patients and 15.0 months for control patients. At the last postoperative visit, mean intraocular pressures were 15.4 +/- 3.7 mm Hg and 15.0 +/- 5.0 mm Hg, respectively (P = .45, Student's unpaired t-test). Both groups showed comparable visual outcome (20/40 or better in 31 of 38 5-fluorouracil-treated patients [82%] vs 32 of 36 control patients [89%]) and a decrease in number of medications needed, 0.7 +/- 1.0 and 0.7 +/- 0.9, respectively (P = .96, Mann Whitney U test). Thus, 5-fluorouracil administered as in our study did not seem to have any effect in primary trabeculectomy combined with phacoemulsification and posterior chamber intraocular lens implantation. PMID- 8238221 TI - Effects of ocular carteolol and timolol on plasma high-density lipoprotein cholesterol level. AB - Fifty-eight healthy, normolipidemic adult men participated in a prospective, masked, randomized crossover study designed to compare the effects of two topical nonselective beta-adrenergic antagonists, carteolol and timolol, on plasma high density lipoprotein cholesterol levels. Two eight-week treatment periods were separated by an eight-week drug-free period. Carteolol 1.0% or timolol 0.5% was used, one drop twice daily, in both eyes without nasolacrimal occlusion. Fresh plasma was assayed for levels of total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, and apolipoproteins A-I and B-100. With indistinguishable effects on intraocular pressure, carteolol and timolol induced different (P = .013) decrements in high density lipoprotein cholesterol levels. Carteolol treatment decreased high density lipoprotein cholesterol levels by 3.3% (-0.04 mmol/l) and raised the ratio of total to high-density lipoprotein cholesterol levels by 4.0% (0.15 unit); timolol treatment decreased high-density lipoprotein cholesterol levels by 8.0% (-0.10 mmol/l) and raised the ratio of total to high-density lipoprotein cholesterol levels by 10.0% (0.37 unit). There was no differential drug effect on the other lipid variables measured. Ocular nonselective beta-adrenergic antagonist therapy can produce clinically relevant decrements in high-density lipoprotein cholesterol levels in healthy men. PMID- 8238222 TI - A prospective trial of intraoperative fluorouracil during trabeculectomy in a black population. AB - In a study population of black Africans with advanced glaucoma in Ghana we conducted a prospective study of intraoperative 5-fluorouracil alone. Eyes undergoing trabeculectomy were randomly selected either to receive or not receive a single intraoperative application of 5-fluorouracil (50 mg/ml for five minutes). Fifty-five eyes had a mean follow-up of 282 days (minimum, 92 days). Twenty of 24 eyes (83%) in the 5-fluorouracil group vs 12 of 31 eyes (39%) in the control group had postoperative intraocular pressure of 20 mm Hg or less with or without medical therapy (P = .01). Eleven of 24 eyes (46%) in the 5-fluorouracil group and five of 31 eyes (16%) in the control group had intraocular pressure of 15 mm Hg or less (P = .02). Without medical therapy, 17 of 24 eyes (71%) in the 5 fluorouracil group and ten of 31 eyes (32%) in the control group had intraocular pressure of 20 mm Hg or less (P = .02). The overall complications were similar in the two groups. In this population, intraoperative 5-fluorouracil markedly improved the ability of trabeculectomy to lower intraocular pressure. We recommend that intraoperative 5-fluorouracil be considered in glaucoma surgery with poor prognosis as an alternative to postoperative subconjunctival injections when multiple injections are not feasible. PMID- 8238223 TI - Comparison of the lens opacities classification system II and Lensmeter 701. AB - We examined 447 hypercholesterolemic and 108 normocholesterolemic Eastern Finnish men, aged 42, 48, 54, or 60 years, who were participating in the Kuopio Atherosclerosis Prevention Study. Two newly developed lenticular opacity grading methods, the Lens Opacities Classification System II and Lensmeter 701, were compared. There was fair agreement between the two grading methods for nuclear color and opalescence, but the agreement was weaker for cortical and posterior subcapsular opacities. The Lens Opacities Classification System II grading of nuclear opacities explained 48% (for the right eye) and 32% (for the left eye) of the variation of Lensmeter readings. Attributable fractions were lower for the other types of lens opacities. PMID- 8238224 TI - Collagen shields impregnated with gentamicin-dexamethasone as a potential drug delivery device. AB - We investigated the ability of collagen shields impregnated with gentamicin sulfate and dexamethasone to deliver medication into rabbit eyes. We compared the aqueous humor gentamicin and dexamethasone levels delivered by collagen shield and subconjunctival injection therapy at five time points over a ten-hour period, by using a fluorescence polarization immunoassay and high-pressure liquid chromatography. Our in vitro studies showed that impregnated collagen shields released most of the gentamicin within the first 30 minutes of elution, whereas dexamethasone was released gradually over a ten-hour period. The collagen shields impregnated with gentamicin and dexamethasone produced aqueous gentamicin levels that were significantly lower (P = .014) than those produced by subconjunctival injection therapy at 30 minutes and that were comparable to subconjunctival injection therapy at the other time points. Minimum inhibitory concentrations of gentamicin (approximately 4 micrograms) were observed in aqueous humor within 30 minutes in rabbits that received subconjunctival injections and at one hour in rabbits treated with impregnated collagen shields. The impregnated collagen shields produced aqueous dexamethasone levels that were significantly lower (P = .004) than those produced by subconjunctival injection therapy at one hour, significantly higher (P = .028) than subconjunctival injection therapy at six hours, and comparable to subconjunctival injection therapy at the other time points. Collagen shield delivery of gentamicin-dexamethasone may be comparable to subconjunctival injections and provide an alternative therapy after intraocular surgery. PMID- 8238225 TI - Ultrasound biomicroscopy in the assessment of anterior scleral disease. AB - High-frequency ultrasound biomicroscopy is a new method of examining subsurface anterior segment structures of the eye at microscopic resolution. The sclera has a high internal reflectivity and can be differentiated from the cornea, and overlying and underlying tissue. Using this modality, we examined 18 patients with various manifestations of scleral disease. Localized anterior staphyloma could be differentiated from other causes of a black spot on the scleral surface. Episcleral thickening could be differentiated from involvement of the sclera itself. Different patterns of scleral involvement could be imaged including diffuse low-reflective mottling, low-reflective nodules extending into the scleral substance, and scleral thinning. Scleral thinning could be assessed and quantified. Underlying changes in the vitreous could be detected. Ultrasound biomicroscopy was a useful adjunct to clinical examination in the assessment of anterior scleral disease. PMID- 8238226 TI - Predicted outcome from hypotensive therapy for glaucomatous optic neuropathy. PMID- 8238227 TI - Acquired Brown's syndrome after a double-plate Molteno implant. PMID- 8238228 TI - Use of a Tono-Pen for long-term home tonometry. PMID- 8238229 TI - Endogenous endophthalmitis caused by Bipolaris hawaiiensis in a patient with acquired immunodeficiency syndrome. PMID- 8238230 TI - Prevention of enucleation of two phthisic eyes by removal of extruding silicone scleral buckles. PMID- 8238231 TI - Muscle loss from doxorubicin injections into the eyelids of a patient with blepharospasm. PMID- 8238232 TI - Lithium-induced downbeat nystagmus in a patient with Arnold-Chiari malformation. PMID- 8238233 TI - Pseudotumor cerebri in a 12-year-old boy with Addison's disease. PMID- 8238234 TI - Bilateral Acanthamoeba keratitis and gas-permeable contact lenses. PMID- 8238235 TI - Diagnosis of advanced Fuchs' endothelial dystrophy with the confocal microscope. PMID- 8238236 TI - Conjunctival keratoacanthoma. PMID- 8238237 TI - Combined embolization and surgical treatment of arteriovenous malformation of the orbit. PMID- 8238238 TI - Management of the posterior capsule during pediatric intraocular lens implantation. PMID- 8238239 TI - Irving H. Leopold 1915-1993. PMID- 8238240 TI - Rous-Whipple Award Lecture. Viruses and diseases of the twenty-first century. PMID- 8238241 TI - Epstein-Barr virus in gastric carcinoma. AB - Epstein-Barr virus (EBV) is known to be related to lymphoid tumors and some types of epithelial tumors, including lymphoepithelioma-like gastric carcinoma with marked lymphocytic stroma. In this study, prevalence of EBV involvement in gastric cancer, and characteristics of tumors with such involvement, were investigated by EBV-encoded RNA 1 in situ hybridization applied to paraffin sections, including the tumor and adjacent gastric tissue, from 999 gastric carcinomas observed in 970 consecutive cases from a large Japanese hospital. EBV involvement occurred in 6.9 percent of lesions, a significantly lower proportion than has been observed in a North American series. Involvement was significantly more frequent among males, in tumors in the upper part of the stomach, and in adenocarcinomas of the moderately differentiated tubular and poorly differentiated solid or medullary types. Almost all carcinomas with marked lymphoid stroma were EBV-positive. Positive lesions were characterized by the presence of uniform hybridized signals in almost all carcinoma cells and by their absence from adjacent non-neoplastic tissue. PMID- 8238242 TI - Increased expression of vascular permeability factor (vascular endothelial growth factor) and its receptors in kidney and bladder carcinomas. AB - Vascular permeability factor (VPF), also known as vascular endothelial growth factor, is a secreted protein implicated in tumor-associated microvascular hyperpermeability and angiogenesis. Tumor cells in 11 of 12 renal cell carcinomas expressed high levels of VPF messenger RNA (mRNA) by in situ hybridization, the only exception being a case of the relatively avascular papillary variant. Expression was further accentuated adjacent to areas of necrosis. Both tumor cells and endothelial cells in small vessels adjacent to tumor stained strongly for VPF protein by immunohistochemistry. Endothelial cells did not express detectable VPF mRNA, but did express high levels of mRNA for the VPF receptors flt-1 and KDR indicating that the endothelial cell staining likely reflects binding of VPF secreted by adjacent tumor cells. Three transitional cell carcinomas also labeled strongly for VPF mRNA. These data suggest an important role for VPF in the vascular biology of these two common human malignancies. PMID- 8238243 TI - Epstein-Barr virus in adult T-cell leukemia/lymphoma. AB - Adult T-cell leukemia/lymphoma (ATLL) is a well-known human T-cell lymphotropic virus type-1-related disease. We studied Epstein-Barr virus (EBV) in the tumor cells of ATLL, to investigate the etiological significance of double infection with these viruses. We used polymerase chain reaction and EBV-encoded small RNA-1 in situ hybridization to investigate the presence of EBV and immunohistochemistry to detect EBV-related oncoproteins, such as EBV-determined nuclear antigen-2 and latent membrane protein. Polymerase chain reaction performed on DNA of frozen specimens from 96 cases of ATLL revealed that the tumor tissue from 21 cases contained EBV DNA. EBV-encoded small RNA-1 in situ hybridization performed on the paraffin sections of the polymerase chain reaction-positive cases indicated EBV in the nuclei of ATLL tumor cells in 16 cases, nine of which were in the pleomorphic nuclei. Latent membrane protein was also detected in the cytoplasm of ATLL tumor cells in 15 cases, and EBV nuclear antigen-2 was observed in the nuclei of ATLL tumor cells in 11 cases. We conclude that EBV was present within tumor cells in about 17% of cases with ATLL and expressed EBV oncoprotein in the tumor cells. It is hypothesized that EBV and human T-cell lymphotropic virus-1 may infect the same T cells in early life and may play a role in the oncogenesis of ATLL. PMID- 8238244 TI - Epstein-Barr virus infection and bcl-2 proto-oncogene expression. Separate events in the pathogenesis of Hodgkin's disease? AB - The present study was undertaken to investigate whether Epstein-Barr virus-(EBV) encoded latent membrane protein (LMP) induces the expression of BCL-2 in Hodgkin and Reed-Sternberg (HRS) cells of Hodgkin's disease (HD) and thereby provide a possible mechanism for the role of EBV in the pathogenesis of this disease. Fifty three cases of HD were studied for the presence of EBV using EBV-encoded RNA in situ hybridization and LMP immunohistochemistry. Immunostaining for BCL-2 on paraffin material was performed using microwave treatment of tissue sections before the application of the primary monoclonal antibody. EBV was located in HRS cells in 16 cases (30%). All cases that were EBV-encoded RNA in situ hybridization positive, also expressed LMP. BCL-2 expression in HRS cells was detected in 16 cases (30%), but only two of these were also EBV-positive. In both of these cases, only occasional HRS cells expressed BCL-2, in contrast to LMP, which was detected in nearly all such cells. BCL-2 staining was predominantly cytoplasmic with some membrane pattern. These results demonstrate that BCL-2 expression can be detected in HRS cells in routinely processed HD tissue and that whereas EBV does not induce the expression of BCL-2 in HD, BCL-2 may have a role in the pathogenesis of EBV-negative cases of HD. PMID- 8238245 TI - In situ hybridization of HIV-1 RNA in retinal vascular wall. AB - In situ hybridization of human immunodeficiency virus-1 (HIV-1) has been performed on eight eyes from eight distinct acquired immune deficiency syndrome patients (three cases had a normal fundus examination and five presented with cytomegalovirus retinitis). The eyes were removed at autopsy and frozen immediately. Contiguous 10-mu cryostat sections were obtained and tested with a HIV probe labeled by nick-translation with [35S]-ATP. HIV-1 RNA was detected in the retina of two acquired immune deficiency syndrome patients. The first positive case presented with typical ophthalmological and histopathological cytomegalovirus retinitis, the second one was not related to cytomegalovirus, according to clinical or histopathological classical criterias. HIV-1 was localized in retinal vascular walls. This shows that there is an active replication of HIV in retina of some acquired immune deficiency syndrome patients. PMID- 8238246 TI - High expression of the Epstein-Barr virus latent protein EB nuclear antigen-2 on pyothorax-associated lymphomas. AB - Pyothorax-associated lymphoma (PAL) is a rare tumor associated with long-standing tuberculous pyothorax. Most of these lymphomas are B-cell lymphomas of high-grade malignancy. Over 50 cases have been reported in Japan, but no cases have been described in Western countries. Its pathogenesis remains unknown. We studied immunohistologically the expression of Epstein-Barr virus- (EBV) encoded latent gene products, EB nuclear antigen-2 and LMP-1, in four cases of PAL. Fifty B-cell lymphomas unrelated to pyothorax, and five EBV-bearing lymphoblastic tumors produced in severe combined immune deficient mice (severe combined immune deficient-EBV+ tumors) were also studied as controls. Marked expression of EB nuclear antigen-2 was demonstrated on all four PALs. LMP-1 was also present in all cases, but both the staining intensity and the number of stained cells remained less than on severe combined immune deficient-EBV+ tumors. Neither EB nuclear antigen-2 nor LMP-1 was observed in the 50 control B-cell lymphomas. Additional molecular genetic analysis revealed that EBVs are incorporated into each PAL clonally. These results confirm the definite association of EBV with PALs, although the significance of weak expression of LMP-1 awaits further study. PMID- 8238247 TI - Dual inhibition of VLA-4 and LFA-1 maximally inhibits cutaneous delayed-type hypersensitivity-induced inflammation. AB - Lymphocytes express surface receptors that mediate adhesion to endothelial cells and control T cell migration into inflammatory sites. Lymphocyte VLA-4 and LFA-1 mediate adhesion to cytokine-activated endothelium, but the contribution of these molecules to in vivo migration and lymphocyte mediated inflammation is not clear. Here we show that both VLA-4 and LFA-1 contribute to not only lymphocyte adhesion but to in vivo lymphocyte migration in the rat and that nearly complete inhibition of lymphocyte accumulation is observed when both integrins are blocked. Furthermore, inhibition of delayed-type hypersensitivity-induced inflammation, as quantified by skin induration and fibrin deposition, is observed with either anti-VLA-4 or anti-LFA-1, but much stronger inhibition is observed with a blockade of both integrins. Thus, dual inhibition of the VLA-4 and LFA-1 pathways is required for a maximal anti-inflammatory effect in some types of T cell-mediated inflammation. PMID- 8238248 TI - Detection of the (11;22)(q24;q12) translocation of Ewing's sarcoma and peripheral neuroectodermal tumor by reverse transcription polymerase chain reaction. AB - Ewing's sarcoma and the related primitive neuroectodermal tumor (PNET) share a unique and specific t(11;22)(q24;q12) chromosomal translocation. The breakpoints have recently been cloned and shown to involve the EWS gene on chromosome 22 and the FLI-1 gene on chromosome 11. Translocation results in the fusion of these genes on the der(22) chromosome, resulting in the production of a novel chimeric EWS/FLI-1 message. Using oligonucleotide primers derived from EWS and FLI-1 complementary DNAs, we were able to amplify a specific fusion transcript from 18 of 18 cases containing t(11;22) and 10 of 14 cases of Ewing's sarcoma/PNET that had unsuccessful cytogenetics. No EWS/FLI-1 fusion transcripts were detected in five cell lines derived from cases of pediatric sarcomas having a histological diagnosis other than Ewing's sarcoma/PNET. The sensitivity and specificity of this PCR analysis demonstrates the usefulness of this approach for the primary diagnosis of t(11;22)-containing Ewing's sarcoma/PNET and for the detection of metastatic or residual disease. PMID- 8238249 TI - Pulsed-field gel electrophoresis analysis of retinoic acid receptor-alpha and promyelocytic leukemia rearrangements. Detection of the t(15;17) translocation in the diagnosis of acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is characterized cytogenetically by a balanced reciprocal chromosomal translocation t(15;17) (q22;q21). This translocation involves the retinoic acid receptor-alpha (RAR-alpha) on chromosome 17 and the promyelocytic leukemia locus (PML) on chromosome 15 and results in the transcription of novel fusion messenger RNAs. In this study, pulsed-field gel electrophoresis (PFGE) was applied to the detection of the t(15;17) translocation in twenty-six clinical specimens cytologically diagnosed by French-American British criteria as APL. This technique could readily be applied to both fresh and nonviably frozen tumor samples. In 24 of 26 samples, rearrangements of the PML and RAR-alpha, loci could be detected by Southern blotting after digestion with MluI and BssHII. Furthermore, co-migration of the rearranged fragments, detected by hybridization to probes for the PML and RAR-alpha genes, demonstrated that these loci were juxtaposed. The translocation was detected in specimens at the time of initial diagnosis, on differentiation therapy with retinoic acid and at the time of relapse. The diagnostic accuracy was compared to cytogenetics and the reverse transcriptase-polymerase chain reaction for the novel PML-RAR-alpha fusion transcript. The samples from two patients were negative by all three diagnostic methods, and both of these patients failed to respond to all-trans retinoic acid. In the other 24 APL samples, cytogenetics was positive in only 76.9% of the cases, whereas both reverse transcriptase-polymerase chain reaction and PFGE methods detected the translocation in 100% of the cases. Thus, PFGE can readily detect the t(15;17) translocation in both viable and nonviable clinical specimens and can improve the diagnostic accuracy of morphology and cytogenetics in APL. In contrast to conventional electrophoresis based on rearrangement of RAR alpha, the ability to demonstrate directly co-migration of the PML and RAR-alpha loci enables this method to distinguish the t(15;17) translocation from variant translocations such as the t(11;15). Because PFGE can be performed on nonviable, frozen tumor samples, it could be diagnostically useful in APL when the RNA-based reverse transcriptase-polymerase chain reaction cannot be performed. PMID- 8238250 TI - Hepatic morphology and iron quantitation in perinatal hemochromatosis. Comparison with a large perinatal control population, including cases with chronic liver disease. AB - We compared hepatic morphology, hepatocellular siderosis, extrahepatic parenchymal siderosis, and (by chemical assay of liver and spleen) the amount of elemental iron and copper in 12 cases of perinatal hemochromatosis (PH) with 119 perinatal controls. Controls were subgrouped according to diagnoses based on clinical and autopsy findings; 37 had chronic liver disease, either hepatic fibrosis (17) or cirrhosis (20). Graded semiquantitatively, hepatocellular siderosis varied widely among controls, and some showed more than PH cases. By chemical assay, total hepatic iron in PH cases was not significantly greater than in any control group except the preterm. Therefore, our findings do not support an etiological role for iron in PH. Its distinctive hepatic morphology seems related to onset of liver disease during fetal life, when periportal hepatocytes normally contain hemosiderin (as in 71 of 82 controls without chronic liver disease). Environmental agents (such as hypoxia, virus, drug) that could damage a fetal liver would usually damage other fetal organs as well. They would be unlikely to recur in a subsequent pregnancy and thereby account for PH occurring in siblings. In initiating PH, therefore, putative environmental agents may need to interact with a factor or factors intrinsic to the developing fetal liver. PMID- 8238251 TI - Deficient differentiation of mast cells in the skin of mi/mi mice. Usefulness of in situ hybridization for evaluation of mast cell phenotype. AB - The staining property of skin mast cells changed from Alcian blue+/berberine sulfate- to Alcian blue+/berberine sulfate+ in the skin of normal (+/+) and Wv/Wv mice. In contrast, this change did not occur in the skin of mi/mi mice. Heparin content and histamine content per a mi/mi skin mast cell were estimated to be 34% and 18% those of a +/+ skin mast cell, respectively. The low heparin content of mi/mi skin mast cells seemed to be consistent with the Alcian blue+/berberine sulfate- staining property. Expression of genes encoding mast cell-specific proteolytic enzymes was examined by Northern blotting and in situ hybridization. Messenger RNA of mast cell carboxypeptidase A was expressed most of all by +/+, Wv/Wv, and mi/mi skin mast cells, but mRNA of mouse mast cell protease (MMCP)-6 was expressed by approximately a half of +/+ and Wv/Wv skin mast cells and by only 3% of mi/mi skin mast cells. A significant amount of MMCP-2 mRNA was not expressed in the skin of all +/+, Wv/Wv and mi/mi mice. This shows the presence of at least three phenotypes in skin mast cells of mice: berberine sulfate+/MMCP 6+, berberine sulfate+/MMCP-6-, and berberine sulfate-/MMCP-6-. The in situ hybridization of mRNA of mast cell-specific proteolytic enzymes seemed to be useful to describe abnormalities of mast cell differentiation in the skin of mi/mi mice. PMID- 8238252 TI - Extensive complement activation in hereditary porcine membranoproliferative glomerulonephritis type II (porcine dense deposit disease). AB - Massive glomerular deposits of C3 and the terminal C5b-9 complement complex (TCC), but no immune complex deposits were detected by immunofluorescence in porcine membranoproliferative glomerulonephritis type II. TCC deposits were always observed with concomitant deposits of vitronectin (S-protein) in membranoproliferative glomerulonephritis, in contrast to a piglet with mesangial glomerulopathy where TCC was present without vitronectin co-deposition. Enzyme immunoassays revealed extensive systemic complement activation in 1-week-old affected piglets, observed by low plasma C3 (about 5% of normal) and high plasma TCC (about 10 x normal). Affected piglets revealed some plasma complement activation already at birth, 3 to 4 weeks before recognizable clinical disease. It is concluded that porcine membranoproliferative glomerulonephritis represents a nonimmune complex-mediated glomerulonephritis caused by unrestricted systemic complement activation with C3 consumption, TCC formation, and glomerular trapping of complement activation products. A pathogenetic mechanism of a defective or missing complement regulation protein is suggested. PMID- 8238253 TI - Temporal expression of autocrine growth factors corresponds to morphological features of mesangial proliferation in Habu snake venom-induced glomerulonephritis. AB - Habu snake venom induces an accelerated mesangial proliferative glomerulonephritis that follows a predictable course from early capillary aneurysms to micronodules comprised of confluent mesangial cells within 72 hours. We examined morphologically the course of mesangial cell proliferation and correlated it with the expression of messenger (m) RNA encoding two peptide growth factors, platelet-derived growth factor (PDGF) A and B chains and transforming growth factor-beta (TGF-beta). Rats were uninephrectomized and 24 hours later injected with Habu snake venom or saline. Kidney cortex and isolated glomeruli were obtained 24, 48, and 72 hours later for histological assessment, preparation and Northern analysis of mRNA, and immunohistochemical localization of PDGF using a polyclonal antibody that recognizes A and B chains. Maximal expression of PDGF B chain mRNA occurred at 24 hours and before the onset of mesangial cell proliferation; whereas maximal expression of PDGF A chain and TGF beta mRNA occurred at 48 hours and during active mesangial cell proliferation. Expression of TGF-beta mRNA persisted at 72 hours at a time when PDGF A chain declined and PDGF B chain was not expressed compared to uninephrectomy and saline controls and at a time when mesangial cells within lesions reached confluence and proliferation subsided. PDGF protein localized in glomerular lesions associated with platelets at 24 and 48 hours and within mesangial cells at 48 and 72 hours. These results agree with the known roles of PDGF and TGF-beta as positive and negative modulators, respectively, of mesangial cell growth in vitro and suggest that a relative balance of the expression of these factors may operate in glomerular disease in vivo. PMID- 8238254 TI - Microvascular pericytes express platelet-derived growth factor-beta receptors in human healing wounds and colorectal adenocarcinoma. AB - The expression of platelet-derived growth factor- beta (PDGF-beta) receptors in the microvasculature of human healing wounds and colorectal adenocarcinoma was investigated. Frozen sections were subjected to double immunofluorescence staining using monoclonal antibodies (MAbs) specific for pericytes (MAb 225.28 recognizing the high-molecular weight-melanoma-associated antigen, expressed by activated pericytes during angiogenesis), endothelial cells (MAb PAL-E), laminin, as well as PDGF-beta receptors (MAb PDGFR-B2) and its ligand PDGF-B chain (MAb PDGF 007). Stained sections were analyzed by computer-aided imaging processing that allowed for a numerical quantification of the degree of colocalization of the investigated antigens. An apparent background colocalization, varying between 23 and 35%, between markers for cells not expected to co-localize was recorded. This background could be due to limitations of camera resolution, to out-of-focus fluorescence, and to interdigitations of the investigated structures. In all six tumor specimens, co-localization of PDGF-beta receptors and PAL-E was not different from the background co-localization, whereas that of PDGF-beta receptors and high-molecular weight-melanoma-associated antigen was significantly higher with mean values between 57 and 71%. Qualitatively, the same pattern was obtained in the two investigated healing wounds. PDGF-B chain did not co-localize with either PAL-E or high-molecular weight-melanoma-associated antigen, but PDGF B chain-expressing cells were, however, frequently found juxtaposed to the microvasculature. The expression of PDGF-beta receptors on pericytes in activated microvessels and the presence of PDGF-B chain-expressing cells in close proximity to the microvasculature of healing wounds and colorectal adenocarcinoma is compatible with a role for PDGF in the physiology of the microvasculature in these conditions. PMID- 8238255 TI - p53 gene mutations and MDM2 amplification are uncommon in primary carcinomas of the uterine cervix. AB - The p53 gene is the most frequently altered gene known thus far in a wide variety of human cancers. Inactivation of p53, either through mutation or through interaction with the human papillomavirus (HPV) E6 oncoprotein, is a characteristic feature of all cervical carcinoma cell lines that have been studied. These findings suggest that p53 inactivation is required for cervical carcinoma development and that HPV infection and p53 mutation may be mutually exclusive. We have studied the p53 gene in 35 primary cervical carcinomas. DNA sequence and single strand conformational polymorphism analyses were used to evaluate p53 in 27 squamous carcinomas (25 HPV-positive) and eight adenocarcinomas (four HPV-positive). A missense mutation of p53 was observed in one HPV 16-positive squamous carcinoma, demonstrating that p53 mutations can occur in combination with HPV infection. The HPV-negative tumors all lacked p53 gene mutations. The absence of p53 mutations in HPV-negative cases prompted an assessment of tumors for MDM2 gene amplification. The MDM2 gene encodes a p53 binding protein and has been found to be amplified in some human tumors lacking p53 mutations. MDM2 amplification was not identified in any of the tumors we examined, including four HPV-negative cases. Our findings show that HPV infection and p53 gene mutation are not mutually exclusive and suggest that many HPV negative carcinomas may arise via a pathway independent of p53 inactivation. PMID- 8238256 TI - Macrophages and multicellular tumor spheroids in co-culture: a three-dimensional model to study tumor-host interactions. Evidence for macrophage-mediated tumor cell proliferation and migration. AB - In a new co-culture model involving multicellular tumor spheroids and different phenotypes of human macrophages, we studied the effects of the latter on migration and proliferation of the human colon carcinoma cell line, HRT-18. The macrophage phenotypes are detectable with monoclonal antibodies and are inducible in culture. 12-O-tetradecanoyl-phorbol-13-acetate-activated macrophages are associated with the phenotype 27E10, which is an acute inflammatory macrophage. The glucocorticoid-induced macrophage phenotype RM3/1 is associated with the down regulation of inflammation. The phenotype resembling the mature resident macrophage termed 25F9 arises spontaneously in prolonged culture. It could be shown that inflammatory macrophages are localized at invasive areas of the tumor host interface of colorectal carcinoma, whereas resident and anti-inflammatory macrophages were found in the central tumor region or at well-bordered areas of the tumor-host interface. The results obtained with this co-culture model show that 27E10-associated macrophages stimulate tumor cell migration and inhibit tumor cell proliferation. RM3/1 had only a slight inhibiting effect on proliferation and a slight promoting effect on migration. The 25F9-positive macrophage-stimulated tumor cell proliferation and inhibited migration completely. This investigation indicates that this in vitro system is useful for studying different macrophage effects on tumor cells and that indeed proliferation and migration of tumor cells could be influenced in an opposite manner by different types of macrophages. PMID- 8238257 TI - Immunohistochemical characterization of intraepithelial and subepithelial mononuclear cells of the upper airways. AB - The upper airway is the first site of exposure to inhaled antigens and the site of initiation of mucosal immunity to certain antigens; however, the intraepithelial lymphoid populations of this region have not been well characterized. We studied 6-mu frozen tissue sections from tonsils, adenoids, and nasal mucosae using immunohistochemistry and a panel of antibodies to mononuclear antigens to determine whether nasal mucosa contained distinctive populations of mononuclear cells. Intraepithelial lymphocytes (IELs) of nasal mucosa were CD3+, CD8+, and mainly CD5+. Tonsil and adenoid both showed diffuse CD8+ IELs; clusters of CD4+ IELs were associated with B cells within the crypt epithelium. All nasal IELs were uniformly negative for Leu8 (homing receptor analog of Mel14). Scattered Leu8-positive cells were present within tonsil and adenoid crypt epithelium only. Nasal IELs rarely expressed HML1 and were often CD7-, whereas the majority of tonsillar and adenoidal IELs were HML1+ and variably CD7+. In nasal mucosa and in deep submucosa of tonsil and adenoid, 80 to 90% of T cell receptor expression was of alpha/beta type. There was a concentration of gamma/delta T cell receptor-positive cells in intraepithelial and subepithelial zones of tonsil and adenoid, with areas of up to 30% gamma/delta T cell receptor positivity. A population of intraepithelial dendritic cells was identified in all three tissues expressing mononuclear phagocyte system antigens CD14 and KiM1P, but lacking CD1a. Virtually no B cells and no organized subepithelial lymphoid tissue were identified in nasal mucosa. Nasal mucosal lymphoid tissue seems to differ from that of endodermally derived mucosae, tonsil, and adenoids to share similarities with both mucosa-associated lymphoid tissue and peripheral lymph nodes. PMID- 8238258 TI - Immunocytochemical and biochemical characterization of the Heymann nephritis antigenic complex in rat L2 yolk sac cells. AB - Heymann nephritis in the rat is the most widely used model of human membranous glomerulonephritis. Glycoprotein (gp)330, a large (M(r) > 550,000) membrane associated glycoprotein, has been identified as the main antigen in this autoimmune disease. Studies of gp330 and receptor-associated protein (RAP), its 44-kd subunit, have been restricted largely to rat kidney, as no stable cultured cell line has been available that expresses gp330. We have recently identified a rat yolk sac carcinoma cell line (L2) that expresses both gp330 and RAP. In this report, we have carried out detailed morphological, immunocytochemical, and biochemical studies characterizing the biosynthesis and localization of gp330 and RAP in the L2 rat yolk sac cell line. At the electron microscope level, the L2 cells are seen to be attached by cell junctions, and their predominant morphological features include extensive networks of rough endoplasmic reticulum (ER) and numerous clathrin-coated pits found on the cell membrane. By immunocytochemistry, gp330 was localized primarily to clathrin-coated pits at the cell surface, whereas RAP was localized predominantly to the lumen of the rough ER. Pulse-chase experiments indicated that gp330 spends a prolonged time maturing in the ER of L2 cells, as transport of gp330 to the Golgi complex (based on acquisition of endoglycosidase H resistance) is slow (t1/2 = 90 to 120 minutes). Gp330 reached the L2 cell surface beginning at 2 hours after synthesis, where it could be detected by cell surface immunoprecipitation. RAP was found to be an N linked glycoprotein, and it remained endoglycosidase H-sensitive up to 4 hours after synthesis. Co-precipitation and co-sedimentation experiments demonstrated that gp330 and RAP form a large heterodimer (M(r) approximately 669,000) immediately after biosynthesis and are further assembled into a large hetero oligomer in the ER. These findings demonstrate that the localization and the kinetics of assembly of gp330 and RAP into the Heymann nephritis antigenic complex are similar in both L2 cells and rat kidney. They also provide new information on the intracellular processing of these two molecules and their delivery to the cell surface. Thus, the L2 cell system should facilitate further characterization of the functions and interactions of gp330 and RAP, which may shed light on the cellular and molecular mechanisms of Heymann nephritis. PMID- 8238259 TI - Induction of different types of glomerulonephritis by monoclonal antibodies derived from an MRL/lpr lupus mouse. AB - MRL/Mp-lpr/lpr(MRL/lpr) lupus mice develop glomerulonephritis in which the histopathological manifestations of the disease are characterized by diffuse cell proliferative, crescentic, and/or wire loop-like lesions, resembling those of human lupus nephritis. Although these lesions are thought to be mediated by antibodies, little data is available to explain these regular variations in glomerular lesions induced by antibodies at the monoclonal level. We studied glomerular lesions of normal or severe combined immunodeficient mice injected with nephritogenic immunoglobulin G3-producing hybridoma clones (2B11.3 and 7B6.8), which we previously established from an unmanipulated MRL/lpr mouse. Both clones caused increased serum levels of immunoglobulin G3 with identical patterns over time and both induced glomerular deposits of immunoglobulin G3 and C3. However, 2B11.3 and 7B6.8 induced glomerular lesions that differed in their histopathological manifestations. The 2B11.3 clone generated cell-proliferative lesions associated with marked Mac-2-positive macrophage infiltrates, but the 7B6.8 clone induced lesions characterized by subendothelial hyaline deposits resembling wire loops. The latter was not associated with significant inflammatory cell infiltrates at any point throughout the progression of the lesion. Thus, our findings suggest that the histopathological variation in glomerulonephritis seen in MRL/lpr mice results from clonally expanded B cell clones that produce nephritogenic antibodies with different pathogenic potencies. PMID- 8238260 TI - Transitional features in human atherosclerosis. Intimal thickening, cholesterol clefts, and cell loss in human aortic fatty streaks. AB - The possible transition from a subset of fatty streaks to fibrous plaques in human atherosclerosis has long been postulated, but transitional features in lesions have rarely been demonstrated. We examined human aortic fatty streaks to determine whether significant tendencies toward intimal thickening and toward deep extracellular lipid deposition might be found. To provide accurate ultrastructural assessment of lipid, tissues were processed by new electron microscopic cytochemical techniques. Unilateral fatty streaks exhibited a 60% increase in intimal thickness when compared to contralateral control tissue. Fat droplets in intimal cells accounted for approximately half of the increase; nonfat portions of cells and extracellular matrix accounted for the remainder. Six of 32 fatty streaks (19%) contained cholesterol clefts, which were found in the musculo-elastic (deep) layer of the intima or in the tunica media. Volume fractions occupied by cells in deep intima were reduced when cholesterol clefts were evident, suggesting loss of cells in early core regions. Light and electron microscopy showed structures consistent with lipid-rich core regions in lesions with cholesterol clefts and in a few lesions without cholesterol clefts. The findings of intimal thickening, core region formation, and disappearance of intimal cells constitute new evidence that some fatty streaks are progressive lesions and sites of eventual fibrous plaque development. The findings also suggest that the lipid-rich core region does not originate primarily from the debris of dead foam cells in the superficial intima, but instead arises from lipids accumulating gradually in the extracellular matrix of the deep intima. PMID- 8238261 TI - Accumulation of abnormal prion protein in mice infected with Creutzfeldt-Jakob disease via intraperitoneal route: a sequential study. AB - We immunohistochemically studied the location of abnormal prion protein in the central nervous system and visceral organs at the clinical and preclinical stages of mice infected with Creutzfeldt-Jakob disease via intraperitoneal route. Abnormal prion protein was diffusely distributed in the central nervous system. The sequential study showed that its stainings were first detected 120 days after inoculation, were found in all mice after 180 days, and were the most intense and widespread after 270 days. There was no restricted involvement at the early stages nor rostrally dominant distribution of the stainings that had been found in mice infected via intracerebral route. Abnormal prion protein was also located in the follicular dendritic cells in the spleen, lymph nodes, intestinal Peyer's patch, and thymus. Its stainings were first detected in the spleen, lymph nodes, and Peyer's patch 14 or 30 days after inoculation. In the thymus, however, the stainings were first detected after 210 days in the germinal centers formed in the medulla. PMID- 8238262 TI - Feline immunodeficiency virus predisposes cats to acute generalized toxoplasmosis. AB - This study was designed to examine the effects of a pre-existing, clinically asymptomatic feline immunodeficiency virus (FIV) infection on a primary challenge with Toxoplasma gondii. Parenteral challenge of FIV-infected cats with tachyzoites of the ME49 strain of T. gondii caused a precipitous drop in all lymphocytes (CD4+, CD8+, and B cells) and generalized severe toxoplasmosis. The predominant postmortem lesions included acute and often fatal interstitial pneumonia, dominated histologically by macrophages, and multifocal to coalescing hepatic necrosis. Immunohistochemistry revealed numerous T. gondii antigen and tachyzoites in macrophages and other cell types in the lung lesions. The proliferative response of peripheral blood mononuclear cells to specific (T. gondii antigen) and nonspecific (Concanavalin A) mitogens was defective in the dually infected cats, suggesting marked immunosuppression. In contrast to the dually infected cats, cats infected only with T. gondii developed a transient, mild clinical disease characterized by anorexia, lethargy, and multifocal chorioretinitis. Lymphocyte changes in T. gondii-infected cats included an early pan-lymphopenia followed by reestablishment of all lymphocyte subset profiles. These cats also showed a reduced proliferative response to Concanavalin A at 1 week after challenge, but a measurable in vivo response to T. gondii antigens, as evidenced by in vitro lymphocyte proliferation in the absence of a mitogenic stimulus. These results show that infection of cats with FIV-NCSU, markedly enhances their susceptibility to a primary T. gondii infection and provides a model to study the mechanisms of the underlying immunological defect(s) occurring early after HIV infection that may predispose individuals to development of acquired immunodeficiency syndrome and associated diseases. PMID- 8238263 TI - Effect of thrombin on maturing human megakaryocytes. AB - Thrombin causes platelet activation and secretion. In some nucleated cells, it is mitogenic. In this study, we have investigated how human megakaryocytes (MKs) respond to this agonist and whether the response depends on the maturation stage. MKs were cultured from bone marrow precursors in liquid culture in the presence of normal plasma. To determine whether thrombin can activate MKs, 14-day MK cultures were incubated with thrombin for 5 minutes, and cells were studied by electron microscopy, either by standard techniques or after embedding in glycol methacrylate for immunoelectron microscopy. Ultrastructural examination of thrombin-treated MKs revealed dramatic morphological changes reminiscent of those found in platelets, including shape change and organelle centralization that involved immature as well as mature cells. MKs were also able to secrete alpha granule proteins in the dilated cisternae of the demarcation membrane system, as shown by immunogold staining for thrombospondin and glycoprotein Ib. These changes were rapid (less than 5 minutes) but despite them, MKs remained viable for more than 24 hours. To determine whether thrombin has a mitogenic activity, it was added to the culture of MKs from day 3 to day 10 of culture at concentrations varying from 0.1 to 10 U/ml. Cells were subsequently studied by a double staining technique using flow cytometry to determine MK number and ploidy. No changes were observed in these two parameters, showing that thrombin is not mitogenic for MKs at the concentrations used. In conclusion, this study confirms for human MKs previous observations made about guinea pig MKs (Fedorko et al, Lab Invest 1977, 36:32). In addition, it demonstrates that immature MKs are able to respond to thrombin and that more mature cells can secrete alpha-granule proteins into the demarcation membrane system, which is in continuity with the extracellular space. This phenomenon may have implications for pathological states such as myelofibrosis formation and for megakaryopoiesis autocrine regulation. PMID- 8238264 TI - Unpublished letters from Prosper Meniere: a personal silhouette. AB - It is ironic that, while the term "Meniere's disease" is becoming more and more well known today, published accounts of a biographic nature are scarce for the man for whom the eponym was devised: Prosper Meniere. As we are obliged to turn to only a few avenues for this sort of information, we are fortunate to discover new sources of original materials to augment the published account of the life and character of Meniere the man. One such unpublished source that has recently come to light is a cache of letters from the pen of Meniere that was preserved for the period extending from the early 1850s up to the time shortly preceding the author's death a decade later; and it is these letters that form the basis of this inquiry into the mind and character of Meniere, the brilliant otologist and man of letters. This particular body of letters, while being for the most part of an admittedly social nature, is of particular interest not only for the occasional references Meniere makes to the ear or to his deaf-mute patients, but also because of Meniere's epistolary style, and because of their content in general. For this reason the letters have been analyzed primarily with an eye to three categories of interest. The first category is the professional life of Meniere, which for the purpose of this paper will be understood to comprise his professional work as a physician, his mention of his patients, of deaf-mutes and their treatment, of his writings in general, and of other pertinent aspects of his medical involvement.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238265 TI - Facial nerve function following cerebellopontine angle surgery: prognostic value of electroneurography. AB - Preoperative counseling of patients, regarding facial nerve function following cerebellopontine angle (CPA) surgery, has remained a difficult task. Facial nerve electroneurography (ENoG) or evoked electromyography can detect subclinical neural degeneration. This test was used in 44 patients undergoing CPA surgery, and facial nerve function was followed postoperatively. A normal preoperative ENoG study appeared to predict an 81 percent and 84 percent chance of good (House grade I-II) early and late postoperative facial function, respectively. An abnormal test is not as reliable. The relation of preoperative ENoG results with tumor size and tumor adherence to the facial nerve is investigated. Electroneurography appears to be a useful preoperative test when counseling patients regarding facial function following CPA surgery. PMID- 8238266 TI - Facial nerve function following cerebellopontine angle surgery: prognostic value of intraoperative thresholds. AB - Intraoperative stimulation of the facial nerve during surgery of the cerebellopontine angle greatly aids the surgeon in identification of the nerve. The preservation of the ability to stimulate the facial nerve, at the brain stem following tumor removal, has been shown to correlate with good postoperative facial outcome. Thresholds of facial nerve stimulation recorded in our series, using constant voltage stimulation, showed statistically significant relevance to facial nerve outcome. When the difference between thresholds before and after tumor removal was 0.2 V or less or when the threshold after tumor removal was less than or equal to 0.2 V, a good postoperative facial outcome could be expected. This paper discusses the relevance of stimulation threshold prior to tumor removal to the size of tumor. PMID- 8238267 TI - Partitioning the labyrinth for benign paroxysmal positional vertigo: clinical and histologic findings. AB - The incidence of benign paroxysmal positional vertigo (BPPV) has been documented at 17 percent of all vertiginous patients. Partitioning of the labyrinth is a laser technique for resolution of BPPV. In 1991 Anthony partitioned the posterior semicircular canal of two patients and eliminated their BPPV in 24 hours, which was much sooner than anticipated. This paper presents an animal study that was undertaken to determine why laser partitioning of the posterior semicircular canal resolved BPPV so promptly. The histologic changes 24 hours post partitioning in the guinea pig showed bony semicircular canal fistula creation, and constriction of the membranous semicircular canal. The constriction caused decreased endolymphatic flow and led to BPPV-resolving long-term obstruction of the membranous canal described by Wilpizeski. Also, the clinical study of the application of partitioning to 14 patients with BPPV is described. The technique under investigation provided the resolution of positional vertigo within 7 days in most cases, and within 8 weeks in all uncomplicated cases. Results included some temporary postoperative motion sensitivity for 6-8 weeks, a transient sensorineural hearing loss in 6 of 14 (43%) patients, and a permanent hearing loss in one (7%) insulin-dependent diabetic patient. Location of the partition on the posterior semicircular canal did not affect the resolution of BPPV, the incidence of transient hearing loss, or the motion sensitivity. Partitioning provided prompt, permanent resolution of BPPV and did not put the hearing at risk in nondiabetic patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238268 TI - Tissue-integrated prostheses in the rehabilitation of auricular defects: results with percutaneous mastoid implants. AB - Functional and cosmetic rehabilitation of the severely atretic auricle poses a formidable challenge. Conventional autologous grafts for auricular reconstruction may produce inconsistent results, and revision of failed grafts is often unsatisfactory. Osseointegrated implant systems to retain auricular prostheses provide an alternative approach in rehabilitating patients with severe auricular defects. Fourteen patients were implanted with 30 osseointegrating implants for rehabilitating major auricular defects and were followed for 1 to 3.5 years, yielding 143 implant observations. Symptomatic skin reactions to the percutaneous implant were noted in 3.5 percent of the observations. Implant extrusion did not occur. This evaluation of tissue-integrated prosthetic systems suggests this approach to be an extremely reliable alternative for selected patients with major auricular defects. The technique of implanting the retention unit is readily adapted to atresia repair. PMID- 8238269 TI - Sensing and controlling rotational orientation in normal subjects and patients with loss of labyrinthine function. AB - The disability resulting from uni-or bilateral loss of vestibular function was assessed by exposing subjects, in darkness, to random rotational displacements on a motorized chair away from a "center" position and requiring them to rotate themselves back to center using a "joystick" control. Normal subjects were accurate within 10 to 15 percent over 180-degree rotations. Alabyrinthine subjects failed to sense their rotation to any useful extent. Subjects with unilateral loss of function as a result of surgery produced hypometric responses to ipsilateral rotation and large amplitude, but inaccurate responses to contralateral rotation. This pattern became more symmetric and normometric with adaptation to loss of function. The procedure promises to be a more useful measure of adaptation than assessing vestibulo-ocular reflexes. PMID- 8238270 TI - Use of poly-l-aspartic acid to inhibit aminoglycoside cochlear ototoxicity. AB - Gentamicin is an extremely effective antibiotic against a wide variety of organisms. However, its use is limited by its nephrotoxic and ototoxic effects. Recent studies in rats have shown that poly-l-aspartic acid (PAA) effectively blocks the nephrotoxic effects of the aminoglycosides and does not decrease the antibiotic effectiveness of gentamicin against those organisms tested. A controlled test was undertaken to evaluate the effect of PAA on cochlear ototoxicity. Test solutions were administered to four groups of guinea pigs for 10 days. Group 1, the controls, received distilled water, group 2 received gentamicin, group 3 received PAA, and group 4 received gentamicin plus PAA. Auditory brainstem response thresholds at 20, 16, 8, and 4 kHz were obtained before therapy, after completion of the 10-day course, and 21 days following the completion of therapy. Two-dimensional diffusion assays in human serum were performed to evaluate the effect of PAA on the antimicrobial activity. Histologic evaluation of the cochleae was performed at the conclusion of the experiment. Threshold shifts following 10 days of therapy were not statistically significant in group 4 compared to controls, while the gentamicin group (group 2) was significantly different for all frequencies tested. At 21 days following therapy, group 4 (PAA + gentamicin) maintained significance in the higher frequencies studied. Antimicrobial studies demonstrated that PAA has no effect on the antimicrobial activity of gentamicin and has no antimicrobial effect against Bacillus subtilis when used alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238271 TI - Partial insertion of the nucleus multichannel cochlear implant: technique and results. AB - The goal of cochlear implant surgery is to insert the entire electrode array into the scala tympani. Incomplete insertion is usually attributable to the presence of new bone formation as a result of meningitis. Surgical techniques for dealing with obstruction include various degrees of drilling of the cochlea or alternative placement of the electrode array. We evaluated the postoperative performance of eight such patients implanted with the Nucleus multichannel cochlear implant at NYU Medical Center as well as data from adults and children nationwide. Scores ranging from marginal closed-set speech discrimination to 40 percent on open-set speech recognition tests were obtained. Surgical procedure, placement, and number of electrodes as well as number of channels in use affect the ultimate performance of these patients. PMID- 8238272 TI - Endoscopic transcutaneous mastoidoscopy for evaluation of residual epitympanic/mastoid cholesteatoma. AB - It is generally agreed that patients subjected to an intact canal wall mastoidectomy to excise mastoid/epitympanic cholesteatoma should undergo a "second-look" operation to rule out residual cholesteatoma. The morbidity of this second operation can be significantly reduced with endoscopic mastoidoscopy, when compared to the traditional postauricular (open) incision approach. Details, benefits, and limitations of endoscopic mastoidoscopy are discussed. PMID- 8238273 TI - Bipolar cochlear nerve recording technique: a preliminary report. AB - To preserve hearing during vestibular neurectomy and acoustic neuroma removal, the cochlear nerve must be identified. Present techniques, including monitoring eighth nerve action potentials, help the surgeon identify those maneuvers that increase the risk of nerve injury but do not help in the anatomic identification of the cochlear nerve or the cochlear-vestibular cleavage plane. The purpose of this study was to demonstrate an electrophysiologic method of identifying the cochlear portion of the eighth cranial nerve. A flush-tipped, bipolar electrode recording probe was used to directly record responses to monaural click stimuli from the cochlear nerve but not from surrounding tissue. It was also used to delineate the cochlear-vestibular cleavage plane. Stimulus intensity levels over 25 dB sensation level tended to reduce the accuracy of nerve identification, and lower levels prolonged recording time. This technique and its application to posterior fossa surgery is discussed. PMID- 8238274 TI - Acute and chronic effects of streptomycin applied to the lateral semicircular canal. AB - In developing the technique of selective chemical vestibulectomy, the destructive effect of streptomycin on the hair cells of the inner ear was well established. However, in both animal and human studies, a rapid onset of the drug's vestibular inhibition was observed in addition to a more slowly developing long-term destructive effect. In previous laboratory studies, streptomycin had been given only by a systemic route, and only the chronic long-term effects had been observed. In this study, an isolated semicircular canal was prepared and streptomycin was placed into the perilymph bathing the canal. The spontaneous and evoked afferent nerve activity were recorded prior to streptomycin application, during application, and after a washout period. During streptomycin application, the activity of the semicircular canal was reversibly inhibited in a dose dependent manner. After a sufficient washout period (5-10 min), the preparation had completely recovered from the drug's effect. Thus, there are two phases in the vestibular inhibition by streptomycin: an early reversible phase that subsequently can transform into a later irreversible phase. PMID- 8238275 TI - Tympanoplasty with mastoidectomy: canal wall up procedures. AB - Management of the mastoid in cases of chronic otitis media with cholesteatoma remains controversial. Whether to leave the canal wall up or perform a cavity technique continues to be debated. The author reviewed his personal cases of surgery for chronic otitis media over a 5-year period and studied 108 cases without prior surgery who required tympanoplasty with mastoidectomy for cholesteatoma. Thirty-two percent of the cases were in children 15 years of age and under. Over two thirds of the procedures were canal wall up, and the remainder of patients underwent a canal wall down technique with obliteration. There was little difference in the results between children and adults, with the exception of there being a tendency for a greater degree of ossicular destruction in the children and a greater incidence of residual disease at second stage surgery. There was a 3 percent incidence of recurrent cholesteatoma. It appears that the intact canal wall technique is preferable in both children and adults, when circumstances are favorable. PMID- 8238276 TI - Biointegrated ventilation tube: current status and new developments. AB - The biointegrated (HydroxylVent) ventilation tube has been used over the past 3 years in a variety of clinical cases. The chief indication for its use is unresolving eustachian tube dysfunction and failure of more conventional medical and surgical therapy. The surgical technique and a postoperative management are described. New directions in clinical research include a larger lumen prosthesis and a compound prosthesis of hydroxylapatite with Teflon lining. PMID- 8238277 TI - Radiologic evaluation of multichannel intracochlear implant insertion depth. AB - Postoperative plain film x-rays are necessary in all multichannel cochlear implant patients to confirm intracochlear position, detect possible electrode kinking, and provide a reference if postoperative slippage occurs. In addition, precise documentation of multichannel intracochlear electrode insertion depths is necessary for comparison of speech recognition results among patients and may be of use for future speech processing strategies. In the present study, a method has been devised, using a modified Stenver's view, to more accurately document insertion depths of the electrode array and location of individual electrodes on 50 multichannel cochlear implant patients. Surgical estimates of insertion depth are shown to have great variability in regard to distance along the basilar membrane when compared with x-ray documentation. Additionally, there is preliminary evidence that insertion depth, as determined by x-ray studies, has a strong correlation with open-set speech discrimination. PMID- 8238278 TI - Restoration of useful hearing after microvascular decompression of the cochlear nerve. AB - This report describes a patient with sudden sensorineural hearing loss who was found to have a megadolichoectasia vertebrobasilar system that appeared to be causing compression of the ipsilateral facial and vestibulocochlear nerves. The patient was treated conservatively for 4 months, during which time no hearing returned. He then underwent microvascular decompression of the affected nerves. At surgery, marked compression of the cranial nerves VII-VIII complex and the pons was observed. Postoperatively, the patient experienced a gradual return of useful hearing. We suggest that vascular compression may be a rare, but treatable, cause of sensorineural hearing loss. PMID- 8238279 TI - Lethal fibrosarcoma complicating radiation therapy for benign glomus jugulare tumor. AB - Radiation therapy for temporal bone paragangliomas has been advocated by many authors, but remains controversial. Radiation has little direct effect on the tumor cells, and diminution in tumor size is seldom seen. The main reason radiation therapy is recommended over surgical management of glomus tumors is its purported lower complication rate. We report a case of a lethal radiation-induced fibrosarcoma presenting 15 years following irradiation for a benign glomus jugulare tumor. Radiation-induced malignancies are rare, but are lethal when they occur. In experienced hands, surgical removal of glomus jugulare tumors carries a limited morbidity and virtually no mortality. In our opinion, the possibility of inducing a secondary life-threatening malignancy must be seriously considered when discussing therapeutic options with otherwise healthy individuals who are expected to survive 10 or more years after treatment. PMID- 8238280 TI - Temporal meningiomas presenting as chronic otitis media. AB - Two cases of temporal bone meningiomas presenting as chronic drainage ears are presented. One case was indicated by computer tomographic scan at the time of initial surgery to be limited to the middle ear, but recurred 9 months later with extensive cerebellopontine angle extension and cranial nerve palsies. The tumor in the second case extended from the middle ear to the cavernous sinus, with extensive intradural involvement. Meningiomas involving the middle ear must be considered to have intracranial extension, and magnetic resonance is the imaging study of choice. PMID- 8238281 TI - Shy-Drager syndrome presenting as Meniere's disease. AB - A 59-year-old patient with a history of Meniere's disease is presented. She developed signs of progressive autonomic failure suggesting a diagnosis of Shy Drager syndrome, a rare degenerative neurologic condition of unknown etiology. The most common otolaryngologic presentation of this syndrome is with stridor attributable to abductor paralysis of the vocal cords. The association with endolymphatic hydrops has not been previously described. PMID- 8238282 TI - Welder's spark and facial paralysis. PMID- 8238283 TI - Facial palsy following cochlear implantation. PMID- 8238284 TI - Role of electrical stimulation therapy for Bell's palsy. PMID- 8238285 TI - Cerumen management by audiologists. PMID- 8238286 TI - Proximal femur of Australopithecus africanus from Member 4, Makapansgat, South Africa. AB - A left proximal femur (MLD 46) from Member 4, Makapansgat, South Africa is described and analyzed. It consists of the head, neck, and a small segment of the shaft that extends to just below the lesser trochanter. The femur exhibits degenerative joint disease in the form of marginal osteophyte formation and thus its taxonomic identity has been somewhat obscured. Consideration of all like sized mammalian femora from Makapansgat suggests that the femur is that of either a felid or hominid. Comparison of MLD 46 to femora of extent and extinct felids reveals that MLD 46 does not possess two morphological features that are characteristic of felids, namely a deep, prolonged trochanteric fossa and a high neck-shaft angle. Simple shape variables (ratios) and multivariate analyses consistently place MLD 46 with modern and fossil hominids, and most closely align it with the australopithecines. We conclude that the femur is most reasonably attributable to Australopithecus africanus, which is the only hominid yet identified from Makapansgat. Despite its pathological condition, MLD 46 is the most complete proximal femur known for A. africanus, thereby permitting further morphological comparisons with homologues of A. afarensis and Paranthropus. Marginal osteophytes of mammalian femoral heads characteristically occur in individuals of advanced age, suggesting that MLD 46 may have lived some time with the disease. Finally, MLD 46 is considerably larger than the previously described specimen, Sts 14, from Sterkfontein Member 4. There may be as great a contrast in body size in A. africanus as there is between the large and small specimens of A. afarensis. PMID- 8238287 TI - Brief communication: Gladysvale: first early hominid site discovered in South Africa since 1948. AB - We report here the discovery of fossil hominid teeth at Gladysvale, near Johannesburg in the southern Transvaal. This find makes the site the seventh in South Africa to yield australopithecine remains and the first new early hominid bearing locality to be found in this region since 1948. Apart from the hominid specimens, our excavations at Gladysvale have added appreciably to the abundant Plio-Pleistocene fauna previously recorded from the cave deposit. The fauna indicates that savanna conditions prevailed during deposition of at least part of the fill. Preliminary faunal dating gives an age of deposition of between c1.7 and c2.5 mya. PMID- 8238288 TI - In memoriam: James Norman Spuhler (1917-1992). PMID- 8238289 TI - Body weight prediction in early fossil hominids: towards a taxon-"independent" approach. AB - The choice of a model taxon is crucial when investigating fossil hominids that clearly do not resemble any extant species (such as Australopithecus) or show significant differences from modern human proportions (such as Homo habilis OH 62). An "interhominoid" combination is not adequate either, as scaling with body weight is strongly divergent in African apes and humans for most skeletal predictors investigated here. Therefore, in relation to a study of seven long bone dimensions, a new taxon-"independent" approach is suggested. For a given predictor, its taxonomic "independence" is restricted to the size range over which the body weight-predictor relationship for African apes and humans converges. Different predictors produce converging body weight estimates (BWEs) for different size ranges: taxon-"independent" estimates can be calculated for small- and medium-sized hominids (e.g., for weights below 50 kg) using femoral and tibial dimensions, whereas upper limb bones provide converging results for large hominids (above 50 kg). If the remains of Australopithecus afarensis really belong to one species, the relationship of male (above 60 kg) to female body weight (approximately 30 kg) does not fall within the observed range of modern hominoids. Considering Sts 14 (22 kg) to represent a small-sized Australopithecus africanus, the level of encephalization lies well above that of extant apes. If OH 62 (approximately 25 kg), with limb proportions less human-like than those of australopithecines, indeed represents Homo habilis (which has been questioned previously), an increase in relative brain size would have occurred well before full bipedality, an assumption running counter to current assumptions concerning early human evolution. PMID- 8238290 TI - Response of bone and enamel formation to nutritional supplementation and morbidity among malnourished Guatemalan children. AB - The effects of changes in nutritional and health status upon bone and enamel development are examined in a sample of 63 rural Guatemalan children (24 females, 39 males). The number of ossified hand-wrist centers at 3 years and the number of linear enamel hypoplasias (LEH) in approximately 0-3 year zones of developing teeth were used to monitor the response of bone mineralization and enamel matrix formation to illness and nutritional supplementation. Numbers of ossified centers and LEH were compared across sex, supplementation, and morbidity groups. Enamel matrix secretion responded positively to increased supplementation. Children who received less than 34.25 kcal/day in supplement had more LEH than those who received more supplement. No differences in ossification status were found between supplementation groups. These data suggest that enamel formation may be more sensitive to changes in nutritional status than is bone mineralization. Disruptions of bone and enamel formation were both associated with frequent illness. Children who were ill more than 3.6% of the time had more LEH and fewer ossified hand-wrist centers than children who were less frequently ill. Conclusions regarding relative environmental sensitivity must take into account the specific aspects of dental and skeletal development examined. PMID- 8238291 TI - Independent test of the fourth rib aging technique. AB - The sternal surface of the 4th rib has been suggested as a useful predictor of adult age-at-death (Iscan et al.: Journal of Forensic Sciences 29:1094-1104, 1984; American Journal of Physical Anthropology 65:147-156, 1984). We tested its ability to do so in two tests. In the first, we developed a method of seriation and target age assignment for the 4th rib so that inaccuracy and bias of the method could be assessed by decade. We found that the 4th rib shows characteristic changes in morphology with age and can be included in multifactorial age estimates. In the second test, we applied the phase method developed by Iscan et al. (Journal of Forensic Sciences 29:1094-1104, 1984). The results of our three judges were generally similar to those reported by Iscan and Loth (Journal of Forensic Sciences 31:122-132, 1986). Measures of race differences in 4th rib morphology were included in both tests. Americans of African descent (black) showed a non-significant trend for the rib changes to be delayed compared to Americans of European descent (white). This is in contrast to the work of Iscan et al., which predicted that blacks would show a tendency toward accelerated rib changes compared to whites, especially after the early 30s (phases 5-7) (Iscan et al.: Journal of Forensic Sciences 32(2):452-466, 1987). PMID- 8238292 TI - Molar microwear in extant small-bodied faunivorous mammals: an analysis of feature density and pit frequency. AB - This study quantitatively examined molar microwear in nine species of extant small-bodied faunivorous primates and microchiropterans. Comparative analyses were performed within the food category faunivory, both between hard- and soft object feeding faunivores and between primarily insectivorous and carnivorous taxa. Additionally, microwear in faunivores was compared to that reported in the literature for frugivorous and folivorous primates. The results indicated that although insectivores and carnivores could not be distinguished by microwear analyses, hard-object faunivores (i.e., those that primarily consume beetles or actively comminute bone) can be readily distinguished from soft-object faunivores (i.e., moth, caterpillar, or vertebrate flesh specialists). The hard-object faunivores consistently exhibited greater pit frequencies (in excess of 40%). Furthermore, comparisons of these microwear data on faunivorous mammals to previous work on frugivorous and folivorous primates (Teaford, 1988, pers. comm.; Teaford and Runestad, 1992, pers. comm.; Teaford and Walker: American Journal of Physical Anthropology 64:191-200, 1984) permitted three observations to be made. 1) Faunivores tend to have higher mean feature densities than either frugivores or folivores, although these differences are not consistently statistically distinct. 2) Faunifores and frugivores that feed on hard-objects have comparable mean pit frequencies. 3) Although it is impossible to distinguish faunivores and folivores on the basis of metric analysis of gross molar morphology, this distinction can be made on microwear criteria. Both hard- and soft-object faunivores exhibit much higher mean pit frequencies than primarily folivorous species. PMID- 8238293 TI - Meningeal arterial patterns in great apes: implications for hominid vascular evolution. AB - Arterial meningeal patterns were observed for 100 hemispheres from great ape endocasts (Pan paniscus, Pan troglodytes, Gorilla gorilla, and Pongo pygmaeus). Eight patterns emerged based on the relative contributions to the walls and dura mater of the middle part of the braincase of meningeal arteries that stem from two sources. These arteries enter the braincase through either the orbit (delivering blood from the internal carotid artery) or through the base of the middle cranial fossa (via the middle meningeal artery whose blood comes from the external carotid artery). The three genera of apes manifest different frequencies of the eight patterns, with orangutans highly dependent on orbital meningeal arteries at one extreme, and chimpanzees showing the greatest reliance on the middle meningeal artery at the other. As was the case in an earlier study of rhesus monkeys, there is a trend across the two genera of African apes for increased mean cranial capacity to be associated with increased reliance on the internal carotid artery for supplying the middle portion of the braincase. However, unlike the case for macaques, this trend does not reach statistical significance in African apes. Because it is rare for humans to manifest significant arterial contributions from the orbit to the middle cranial fossa, the comparative data on monkeys, apes, and humans suggest that, during the course of vascular evolution in Homo, the middle meningeal artery eventually took over supply of the entire middle cranial fossa. This hypothesis should be tested in the hominid fossil record. Earlier work on meningeal arterial patterns in apes has traditionally relied on Adachi's system that was determined from humans and focuses on the origin of the middle branch of the middle meningeal artery. As a result, the extensive orbital contributions to the middle portion of the braincase that characterize apes were not recognized and the eight patterns described in this paper were often erroneously assigned to the three patterns that adequately describe only humans. Adachi's system should therefore be abandoned for nonhuman primates and early hominids. A correct understanding of meningeal arterial evolution cannot be achieved until the orbital contributions to the meningeal arteries are recognized and incorporated into an evolutionary study that spans from apes to fossil hominids to living people. PMID- 8238294 TI - Orientation of the masseter muscle and the curve of Spee in relation to crushing forces on the molar teeth of primates. AB - A highly significant correlation is demonstrated between the forward inclination of the superficial masseter muscle and the forward tilt of molar teeth in the sagittal plane, conforming to the posterior end of the curve of Spee. The skulls studied included New and Old World monkeys, anthropoid apes, man, and a selection of other mammals. There was a wide variation between the angles measured in different animals (from 54 degrees to 115 degrees). The tilt of the curve of Spee increases the crush/shear ratio of the force produced on food between the posterior molars. The parallel tilt of superficial masseter maximizes the efficiency with which this muscle contributes to the crush component of the bite force in the sagittal plane. PMID- 8238295 TI - Characterization of amino acid transport in human endothelial cells. AB - The transport of amino acids has been studied in human umbilical vein endothelial cells. Neutral amino acids enter human umbilical vein endothelial cells through three distinct agencies endowed with the characteristics of systems A, ASC, and L. Each system has been studied by evaluating the influx of preferential substrates. The influx of L-proline and 2-methylaminoisobutyric acid occurs through an Na(+)-dependent adaptively regulated trans-inhibited agency identifiable with system A. L-Threonine influx occurs mainly through a distinct Na(+)-dependent trans-stimulated pathway corresponding to system ASC. System L accounts for Na(+)-independent influx of L-leucine. These systems cooperate for the transport of L-glutamine, which is due mainly to system ASC, whereas the component due to the operation of system A increases upon amino acid starvation. No clear evidence was found for a glutamine-specific system ("system N"). Two systems, one Na+ dependent (system XAG-) and the other Na+ independent (system xc ), transport anionic amino acids. L-Arginine influx exhibits a poor dependence on extracellular Na+, whereas it is sensitive to conditions known to change membrane potential and to trans-stimulation by intracellular amino acids. These features are consistent with a process mediated by system y+ and may be of significance for the regulation of the intracellular concentration of L-arginine. PMID- 8238296 TI - Vacuolar-type H(+)-ATPases are functionally expressed in plasma membranes of human tumor cells. AB - Mammalian cells generally regulate their intracellular pH (pHi) via collaboration between Na(+)-H+ exchanger and HCO3- transport. In addition, a number of normal mammalian cells have been identified that express H(+)-adenosinetriphosphatases (ATPases) in their plasma membranes. Because tumor cells often maintain a high pHi, we hypothesized that they might functionally express H(+)-ATPases in their plasma membranes. In the first phase of the present study, we screened 19 normal and tumorigenic human cell lines for the presence of plasmalemmal H(+)-ATPase activity using bafilomycin A1 to inhibit V-type H(+)-ATPase and Sch-28080 to inhibit P-type H(+)-K(+)-ATPase. Bafilomycin A1 decreased pHi in the six tumor cell lines with the highest resting pHi in the absence of HCO3-. Sch-28080 did not affect pHi in any of the human cells. Simultaneous measurement of pH in the cytoplasm and in the endosomes/lysosomes localized the activity of bafilomycin to the plasma membrane in three cell lines. In the second phase of this study, these three cell lines were shown to recover from NH4(+)-induced acid loads in the absence of Na+. This recovery was inhibited by N-ethylmaleimide, bafilomycin A1, and ATP depletion and was not significantly affected by vanadate, Sch-28080, or hexamethyl amiloride. These results indicate that a vacuolar type H(+)-ATPase is expressed in the plasma membrane of some tumor cells. PMID- 8238297 TI - Chloride current activated by swelling in retinal pigment epithelium cells. AB - A membrane conductance activated by cell swelling was characterized in cells of the retinal pigment epithelium (RPE). Manipulations of internal and external Cl concentration revealed that the conductance is permeable to Cl and somewhat permeable to the gluconate anion used for Cl substitution (ratio of gluconate to Cl permeability approximately 0.1). The conductance was blocked by the Cl channel blockers 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and 4-acetamido-4' isothiocyanostilbene-2,2'-disulfonic acid in a manner consistent with open channel blockade. Both the onset and recovery of the Cl current following a transient increase in cell volume were slow. This suggests that activation of the current depends on some internal signal rather than directly on membrane stretch. Experiments to examine a possible role for intracellular Ca concentration ([Ca]i) in regulation of the current demonstrated that an increase in [Ca]i was not involved in the linkage between swelling and Cl current; activation of the current was unaffected by the calcium-buffering conditions, the current could not be activated by large increases in [Ca]i elicited by ionomycin, and no changes in [Ca]i were observed to be associated with swelling. RPE cells normally experience changes in the volume and ionic composition of the extracellular subretinal space during changes in illumination conditions; therefore, the volume-sensitive Cl conductance may play a role in volume regulation in the RPE in response to these extracellular changes and/or in transepithelial fluid transport. PMID- 8238298 TI - Angiotensin II type 1 receptor-mediated stimulation of c-fos gene expression in astroglial cultures. AB - Angiotensin II (ANG II) stimulates plasminogen activator inhibitor 1 (PAI-1) gene expression in astroglial cells prepared from rat brains. In this study, we investigated whether c-fos gene expression may be involved in this cellular action of ANG II. Incubation of astroglial cultures with ANG II caused a time- and dose-dependent transient stimulation of the steady-state levels of c-fos mRNA, with a maximal stimulation of 50-fold observed with 100 nM ANG II within 30 45 min. This stimulation was completely abolished by the presence of the type 1 ANG II (AT1) receptor antagonist losartan but not by the type 2 ANG II receptor blocker PD-123177. Depolarization of brain cell cultures with 50 mM K+ also caused a 100-fold increase in c-fos mRNA levels, an effect partially blocked by losartan. These observations show that AT1 receptor activation stimulates expression of the c-fos gene, which may act as a third messenger in the regulation of cellular actions of ANG II, including PAI-1 gene expression in astroglial cells. PMID- 8238299 TI - A putative H(+)-K(+)-ATPase is selectively expressed in surface epithelial cells of rat distal colon. AB - Recently, a putative distal colon H(+)-K(+)-ATPase alpha-subunit has been identified and characterized (M. S. Crowson and G. E. Shull. J. Biol. Chem. 267:13740-13748, 1992). In the present study, we report the tissue and cell expression of this putative H(+)-K(+)-ATPase. The results indicate that, first, in the gut, the putative H(+)-K(+)-ATPase alpha-subunit is restricted to the distal part of the colon and is predominantly expressed in surface epithelial cells, in marked contrast to the alpha 1-subunit of Na(+)-K(+)-ATPase that is also expressed in glands. These data suggest that the H(+)-K(+)-ATPase alpha subunit is a potential marker for terminal differentiation of distal colon. Second, in the uterus, the putative H(+)-K(+)-ATPase is restricted to the region of the myometrium between the inner and midmuscular zone that is very rich in vascular supply and nerve cells. This striking expression suggests that the H(+) K(+)-ATPase may not be involved in the control of pH and potassium concentration of the uterine fluid but rather in distinct functions of vascular and/or nerve cells. Third, with the use of three independent and different approaches (Northern blot analysis, ribonuclease protection assay, and in situ hybridization), we were unable to detect any significant amount of H(+)-K(+) ATPase transcripts in kidney tissue. Our data suggest that the putative distal colon H(+)-K(+)-ATPase is probably distinct from the kidney isoform. Finally, we report the sequence of a set of degenerate oligonucleotides that are useful to clone alpha-subunits of the Na(+)-K(+)-/H(+)-K(+)-ATPase gene family in different tissues and different species. PMID- 8238300 TI - Angiotensin-mediated phosphatidylcholine hydrolysis and protein kinase C activation in mesangial cells. AB - Angiotensin II (ANG II) in mesangial cells (MC) promotes phosphatidylinositol (PI) hydrolysis resulting in diacylglycerol (DAG)-mediated increases in protein kinase C (PKC) activity. The paucity of MC inositol lipid prompted us to consider whether phosphatidylcholine (PC) could sustain DAG formation. ANG II released choline and increased phosphatidylethanol (PEt) via PC-phospholipase D (PC-PLD). ANG II also stimulated phosphorylcholine consequent to PC-phospholipase C (PC PLC) activation. ANG II-mediated PC hydrolysis augmented DAG for 30 min. PC breakdown was influenced by extracellular Ca2+, because Ni2+ partially inhibited ANG II-induced PEt and obliterated agonist-mediated DAG formation. The consequence of Ca2+ modulation of PC metabolism was investigated by measuring PKC activity. Ni2+ had no effect on early (PI-associated) activation by ANG II at 90 s but obviated translocation from cytosol to the membrane at 10 min. The pathway responsible for PC-associated DAG was studied in PKC downregulated cells. Whereas downregulation prevented PLD-mediated PEt elevation, ANG II-stimulated DAG formation in myristate-labeled cells was unaltered, indicating PC-PLC activation. In summary, ANG II stimulates PC-PLD and PC-PLC in MC. PC-PLD is tightly regulated by PKC, whereas PC-PLC is stringently controlled by extracellular Ca2+. ANG II mediated PC breakdown principally via PC-PLC provides a mechanism for maintaining elevated DAG levels and PKC activation. PMID- 8238301 TI - Phorbol ester sequentially downregulates cAMP-regulated basolateral and apical Cl transport pathways in T84 cells. AB - The effect of phorbol esters on adenosine 3',5'-cyclic monophosphate (cAMP) regulated epithelial Cl- transport was studied in T84 cells, a human colonic cell line that serves as a model for electrogenic Cl- secretion. Preincubation of T84 cell monolayers with phorbol 12-myristate 13-acetate (PMA) caused a time- and dose-dependent inhibition of the net transepithelial secretory response to 10 microM forskolin (half-maximal inhibition at a concentration of approximately 10 nM PMA and a time of 45 min). Similar inhibition was observed with phorbol 12,13 dibutyrate but not the inactive phorbol ester phorbol 12,13-diacetate. Na(+)-K(+) 2Cl- cotransporter activity, assessed by bumetanide-sensitive 86Rb+ uptake, and K+ conductance, assessed by 86Rb+ efflux, were both found to be markedly reduced by PMA with a time course that paralleled the loss of the cAMP-regulated Cl- secretory response. One- and four-hour treatment of T84 cells with 100 nM PMA caused a sustained increase in the membrane-bound fraction of protein kinase C (PKC) but a decrease in total cellular PKC. Although, at these time points, the Na(+)-K(+)-2Cl- cotransporter and K+ efflux pathways were markedly inhibited (associated with inhibition of the forskolin-stimulated transepithelial Cl- secretory response), the activity of the cAMP-regulated Cl- efflux pathway, assessed by 125I-labeled efflux, remained unaffected. With prolonged exposure to PMA (up to 10), the cAMP-regulated Cl- efflux pathway was also eventually inhibited, and transepithelial electrical resistance progressively declined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238302 TI - Alpha-chymotrypsin deregulation of the sodium-calcium exchanger in barnacle muscle cells. AB - To gain insight into the mechanism by which the protease alpha-chymotrypsin (alpha-chym) activates the Na-Ca exchanger in muscle cells we studied 1) the ability of this enzyme to remove the intracellular "catalytic" Ca2+ requirement for activation of all the modes of exchange mediated by the Na-Ca exchanger (i.e., Nao-Cai, Nai-Cao, Nao-Nai, and Cao-Cai, where the subscripts o and i represent extracellular and intracellular, respectively), and 2) the ability of certain monovalent cations to stimulate Cao-Cai exchange after activation of the exchanger by alpha-chym. Barnacle muscle cells were used as models because these cells are so large that they can be internally perfused and voltage clamped. The results show that alpha-chym produces a highly activated Na-Ca exchanger able to operate in all its modes of exchange independently of catalytic Cai. The concentration-dependent effect of alpha-chym was biphasic; maximal activation occurred at 0.5 mg alpha-chym/ml perfusate for 20 min of perfusion at a perfusion rate of 2.5 microliters/min. The results are discussed in terms of the possible effects of alpha-chym on the kinetic modulation of the exchanger. PMID- 8238303 TI - External Ca effect on water permeability, regulatory volume decrease, and extracellular space in barnacle muscle cells. AB - The effect of extracellular Ca2+ (Cao) on sarcolemmal hydraulic water permeability (L'p), regulatory volume decrease (RVD), and extracellular space (ECS) was studied in barnacle muscle cells. Absence or presence of Cao had no effect on L'p [0 Cao = 2.762 +/- 0.098 x 10(-5), and 11 mM Cao = 2.720 +/- 0.222 x 10(-5) cm.kg.s-1 x osmol x 1-kgH2O-1]. Likewise, cells exposed to anisosmotic media (for < 30 min) behaved as osmometers in 0 and 11 mM Cao, showing similar slopes and intercepts in van't Hoff plots. At longer incubation times, however, hyposmotic conditions promoted a Cao-dependent RVD. The relationship between Cao and the percentage of cells responding with RVD to a hyposmotic challenge was sigmoidal (half-maximal Cao = 4.83 mM). The mean rate of RVD (40 nl/min) was independent of the level of swelling in response to hyposmotic challenges. However, the magnitude of RVD increased with larger hyposmotic challenges. Both the presence of Cao and hypotonicity reduced the "apparent" ECS by 47 +/- 6 and 39 +/- 6%, respectively. Three-dimensional reconstruction of autoradiographs of the cells was made to interpret these results. PMID- 8238304 TI - Spatial changes of [Ca2+]i and contraction caused by phorbol esters in vascular smooth muscle cells. AB - In A7r5 smooth muscle cell suspensions, 12-deoxyphorbol 13-isobutyrate (DPB) and phorbol 12,13-dibutyrate (PDB), active phorbol esters for protein kinase C (PKC), increased the intracellular Ca2+ concentration ([Ca2+]i), but 4 alpha-PDB, an inactive phorbol ester, did not. Digital images of the fura 2 fluorescence from single cells revealed that DPB caused elevation of Ca2+ in localized peripheral regions, followed by expansion of this elevated Ca2+ level throughout the cytoplasm. High K+ also induced a dynamic change of [Ca2+]i. DPB and high K+ increased the wrinkles and distortions in the flexible growth surface beneath the cells. In Ca(2+)-depleted media, DPB did not affect [Ca2+]i but substantially increased wrinkles. The increase could be eliminated by staurosporine, a specific inhibitor of PKC. DPB increased the particulate PKC activity in a concentration dependent manner. These results suggest that PKC activation induces cellular contractions through two distinct mechanisms, one dependent on and another independent of the [Ca2+]i increase. PMID- 8238305 TI - Effects of adenosine antagonists on hexose uptake and preconditioning in perfused rat heart. AB - Preconditioning with brief intermittent periods of ischemia has been shown to lessen the detrimental effects of a subsequent sustained (30-60 min) period of ischemia. Because adenosine has been suggested to be the mediator of preconditioning, we were interested in investigating whether adenosine antagonists would block the effect of preconditioning on ionic changes during ischemia. We found that 10 microM of the adenosine antagonist BW-A1433U did not reverse the effect of preconditioning on intracellular pH (pHi). Hearts preconditioned with BW-A1433U had virtually no decrease in pHi during the 30-min sustained period of ischemia; after 30 min of ischemia, the pH in untreated hearts was 5.97 +/- 0.16 compared with 6.52 +/- 0.10 in preconditioned hearts and 6.90 +/- 0.08 in hearts preconditioned plus BW-A1433U. Because anaerobic glycolysis is largely responsible for the fall in pHi during ischemia, we examined the effect of BW-A1433U [and other adenosine antagonists, such as PD 115,199 and 8-cyclopentyl-1,3-dipropylxanthine (CPDPX)] on glucose uptake and phosphorylation during aerobic perfusion using 31P-nuclear magnetic resonance to monitor uptake and phosphorylation of 2-deoxyglucose (2-DG) to 2-deoxyglucose 6 phosphate (2-DG-6-P) when one-half of the glucose in the perfusate was replaced with 2-DG. Uptake of 2-DG-6-P after 15 min was reduced by 66% in the presence of BW-A1433U and 82% in the presence of PD-115,199 as compared with untreated hearts, but was not reduced in the presence of CPDPX. Thus CPDPX was the only adenosine antagonist tested that did not block accumulation of 2-DG-6-P. We also found that CPDPX did not block the beneficial effect of preconditioning on ionic alterations during a sustained 30-min period of ischemia or the improved recovery of function on reflow. PMID- 8238306 TI - Glycosylphosphatidylinositol-specific phospholipase D is localized in keratinocytes. AB - Glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) is abundant in mammalian plasma, but little is known of its cellular and tissue distribution. In this study frozen sections of perfused tissues from adult rats were stained with monoclonal antibodies against GPI-PLD. The most intense staining was observed in the stratified squamous epithelium of the forestomach. Staining was also observed in the esophagus, the tongue, the hard palate, and the skin but not in most other tissues including the columnar epithelium of the stomach or the lower gastrointestinal tract. GPI-PLD expression was also detected in several keratinocyte cell lines. Biochemical assays of glycosylphosphatidylinositol degrading activity using [3H]myristate-labeled variant surface glycoprotein as substrate provided independent evidence for the presence of GPI-PLD. Expression of GPI-PLD by keratinocytes was not affected by culture in serum-free media, indicating that it does not originate by uptake of serum GPI-PLD in the media. These data suggest that keratinocytes are an important site of action of GPI-PLD and possibly a contributor to the plasma GPI-PLD pool. PMID- 8238307 TI - Mutation of a conserved proline residue in the beta-subunit ectodomain prevents Na(+)-K(+)-ATPase oligomerization. AB - A highly conserved sequence motif (4 tyrosines and 1 proline: YYPYY) of the Na(+) K(+)-adenosinetriphosphatase (ATPase) beta 1-subunit ectodomain has been mutagenized to study its possible role in alpha/beta-assembly and sodium pump function. Single as well as double tyrosine mutants (tyrosine to phenylalanine: Y to F) of Xenopus laevis beta 1-subunits are able to associate with alpha 1 subunits and form functional Na-K pumps at the plasma membrane that are indistinguishable from wild-type alpha 1, beta 1-Na-K pumps (as assessed by measurements of ouabain binding, 86Rb flux, Na-K pump current, and activation by external potassium). In contrast, a single proline mutation (proline to glycine: P244G) reduced by > 90% the proper assembly and function of Na(+)-K(+)-ATPase, despite a normal rate of synthesis and core glycosylation. Our data indicate that proline-244 plays a critical role in the proper folding of the beta-subunit and its ability to associate efficiently with the alpha 1-subunit in the endoplasmic reticulum. PMID- 8238308 TI - Amiloride-sensitive sodium conductance in human B lymphoid cells. AB - The Na+ conductance of RPMI 8226 human B lymphoblastoid cells was examined using whole cell patch clamp. When the bath solution contained RPMI 1640 and the pipette solution contained (in mM) 100 potassium gluconate, 30 KCl, 10 NaCl, 0.5 ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), and 20 N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES), as well as < 10 nM free Ca2+, pH 7.25, the mean membrane potential was -58 +/- 4.5 mV (n = 18). Cells were voltage clamped from -160 to +40 mV in 20-mV increments. The inward conductance was 767 +/- 221 pS/10 pF, and the outward conductance was 1,212 +/- 272 pS/10 pF (n = 12). Superfusion with 2 microM amiloride significantly hyperpolarized the cells by 7.4 +/- 2.2 mV (P = 0.004), significantly reduced the inward conductance to 221 +/- 65 pS/10 pF (P = 0.028), but had no effect on the outward conductance (1,294 +/- 236 pS/10 pF, P = 0.820, after amiloride). Next, the pipette and bath solutions were changed to (in mM) 150 sodium gluconate, 0.5 EGTA, and 20 HEPES, as well as < 10 nM free Ca2+, pH 7.25. Under these conditions amiloride significantly reduced (50%, P < 0.05; n = 7) the whole cell currents. When potassium gluconate was substituted for sodium gluconate, amiloride had no effect. Thus amiloride inhibited a Na(+)-specific conductance expressed by B lymphoid cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238309 TI - Simultaneous imaging of intracellular [Ca2+] and pH in single MDCK and glomerular epithelial cells. AB - The interrelationships between changes in intracellular calcium concentration ([Ca2+]i) and intracellular pH in Madin-Darby canine kidney cells and kidney glomerular epithelial cells exposed to various stimuli were analyzed simultaneously using a new design of a fluorescence video microscope. Cells were double labeled with indo 1 and SNARF 1 dyes and were excited simultaneously at 350 and 540 nm. Images at four emission wavelengths were captured simultaneously at 405, 475, 575, and 640 nm at 30 frames/s for the two ratio dyes. SNARF sensitivity to pH between 6.5 and 8.0 was unchanged by [Ca2+]i. The SNARF ratio maps were used to correct the pH-dependent changes in the calculation of local cell calcium. NH4Cl loading produced the expected alkalinization and a concurrent rise in [Ca2+]i. When the NH4Cl was removed and the cells became acidic, a second rise in [Ca2+]i was recorded. Both changes in [Ca2+]i were from intracellular stores since they persisted in the absence of extracellular calcium. The findings demonstrate the need for pH correction of indo 1 recordings. PMID- 8238310 TI - Positive and negative regulation of chloride secretion in T84 cells. AB - This review discusses recent findings regarding the mechanisms and regulation of chloride secretion in the T84 cell line, a widely used model for the study of transepithelial chloride transport, the properties of the cystic fibrosis gene product (cystic fibrosis transmembrane conductance regulator), and the interactions of inflammatory cell types with the intestinal epithelium. First, I review the features of the chloride secretory mechanism in this cell line, both as originally described by Dharmsathaphorn and co-workers and as modified by more recent findings. Second, I cover what is known of the intracellular regulation of the secretory process. Third, I review the ways in which the cell line has been utilized to dissect pathways of immune-epithelial interactions that may contribute to the pathogenesis of inflammatory diarrhea. Finally, I suggest areas of investigation with this cell line that may prove ripe for further study. The physiological and pathophysiological implications of findings obtained with the cell line are also discussed. PMID- 8238311 TI - Receptor-activated currents in mouse fibroblasts expressing transfected bombesin receptor subtype cDNAs. AB - BALB/c 3T3 cells do not normally express receptors for bombesin-like peptides [bombesin (Bn), gastrin-releasing peptide (GRP), and neuromedin B (NmB)]. Transfection of BALB/c 3T3 cells with complementary DNA-encoding GRP receptors or NmB receptors leads to stable expression of functional GRP receptors (GRP Rt) or NmB receptors (NmB Rt), respectively, which are coupled to cell membrane ion channels. Whole cell current analysis using patch electrodes shows that the activation of these newly expressed receptors induces cation conductance increases, most frequently a Ca(2+)-activated plasma membrane K+ conductance. The dose-response (peak-current) relations of both transfected receptor subtypes were sigmoidal and exhibited threshold activation concentration in the picomole range and the saturation of responses to higher concentrations than 10(-8) M. The GRP Rt responded about equally to GRP, NmB, and Bn when compared at equimolar levels, despite their known difference in binding affinity for the three peptides (GRP, Bn > NmB). In contrast, for the NmB Rt, the NmB was more potent than GRP or Bn. Among four GRP/Bn-receptor antagonists tested, the [D-Phe6]Bn(6-13) ethyl ester suppressed GRP Rt responses at low concentrations (10(-7) M). N-acetyl-GRP-(20 26) amide, [Leu13-psi(CH2NH)-Leu14]Bn, and [D-Arg1,D-Phe5,D Trp7,9,Leu11]substance P also blocked GRP Rt responses but at higher concentrations (10(-5) M). However, at these concentrations, these four antagonists had little effect on NmB Rt responses, thereby showing a specificity of these antagonists for the GRP receptors. PMID- 8238312 TI - Emptying and refilling of Ca2+ store in tracheal myocytes as indicated by ACh evoked currents and contraction. AB - Membrane currents and contractions evoked by acetylcholine (ACh) in freshly dissociated canine tracheal myocytes were investigated using the nystatin perforated-patch recording technique. In cells held at -60 mV in the presence of nifedipine, ACh evoked inward current (IACh) and contraction. Caffeine mimicked the effects of ACh. IACh and contractions could be evoked 3-4 min after removing external Ca2+ but were abolished by prolonged exposure to Ca(2+)-free media. Both responses were restored within minutes of reintroduction of Ca2+, even though the cells were held at -60 mV in the presence of nifedipine. IACh and ACh-evoked contractions were also reversibly abolished by continued exposure to caffeine. Cyclopiazonic acid (CPA), a blocker of the sarcoplasmic reticulum (SR) Ca(2+) ATPase, reduced IACh by > 95% within 15 min but had little or no effect on the contractile responses evoked by ACh. IACh was restored after washout of CPA even though cells were held at -60 mV. After depleting the Ca2+ store with the use of CPA, depolarization of the membrane to +10 mV immediately before application of ACh led to a partial restoration of IACh. This restorative effect of depolarization was potentiated by Bay K 8644 and antagonized by nifedipine. In conclusion, IACh and contractions in canine tracheal myocytes are mediated by Ca2+ released from an internal store that can be depleted by prolonged removal of extracellular Ca2+, prolonged exposure to caffeine, or by blockade of the SR Ca(2+)-ATPase. At least two Ca2+ influx pathways appear to contribute to refilling of the internal store: one pathway that is not activated by depolarization or ACh and a second involving dihydropyridine-sensitive voltage activated Ca2+ channels that may be in direct contact with the SR (i.e., conduct extracellular Ca2+ directly into the SR, bypassing the cytosol). PMID- 8238313 TI - Surplus Na+ pumps: how low-K(+)-incubated LLC-PK1 cells respond to K+ restoration. AB - We have previously shown that a pig kidney cell line (LLC-PK1/Cl4) responds to chronic exposure to 0.25 mM extracellular K+ by increasing the beta-, not alpha-, subunit mRNA levels and both alpha- and beta-abundance twofold over control. Our objective in the present study was to determine how the LLC-PK1/Cl4 cells respond when returned to control (5.5 mM) medium. A 1.8-fold increase in ouabain binding established that the induced pumps were expressed at the cell surface following 24-h incubation in low K+. On restoration to 5.5 mM K+, intracellular Na+ and K+ concentrations ([Na+]i and [K+]i, respectively) rapidly returned to control levels within 15 min. The doubled pool size of pumps in the chronic low K+ cells had no significant influence on the rate of ion restoration when compared with the rate in cells acutely exposed to low K+. Despite the rapid return of ions to control values, beta-mRNA levels remained elevated for 2 h, then sharply declined to control levels by 6 h of K+ restoration. From these data, we estimate that the half-life of beta-mRNA is 2-3 h during restoration. alpha-Subunit mRNA remained essentially unchanged from control after return of K+ to the medium and restoration of intracellular ions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238314 TI - Zn(II) transport and distribution in rat spermatids. AB - Zn(II) is an essential trace element. In spermatozoa, Zn(II) modulates metabolism and chromatin condensation. The mechanisms of uptake and distribution of this ion in sperm cells have not been explored. In rat spermatids, our results indicate that 1) 65Zn(II) binds with fast kinetics to a labile, presumably extracellular, compartment. This binding is temperature insensitive and not modified by metabolic inhibitors. 2) Entry of 65Zn(II) in the absence of externally added proteins occurs through a mediated transport system that allows exchange to reach steady state in approximately 15 min at 34 degrees C. 3) Upon entering the cells, 65Zn(II) binds tightly to cellular organelles. 4) Exchangeable Zn(II) bound to cytoplasmic proteins plus free intracellular Zn(II) appears to be < 20% of total exchangeable Zn(II). 5) The intracellular exchangeable Zn(II) compartment is decreased by metabolic inhibitors, showing a direct or indirect link between energy metabolism and cellular Zn(II) levels. 6) 65Zn(II) efflux from rat spermatids is a process with a rate constant of 0.16 +/- 0.13 min-1 at 34 degrees C. This exit rate of Zn(II) is likely to be affected by Zn(II) release from cytoplasmic binding sites or organelles. PMID- 8238315 TI - Acidification of rat liver lysosomes: quantitation and comparison with endosomes. AB - Both lysosomes and endosomes are acidified by an electrogenic proton pump, although studies in intact cells indicate that the steady-state internal pH (pHi) of lysosomes is more acid than that of endosomes. We undertook the present study to examine in detail the acidification mechanism of purified rat liver secondary lysosomes and to compare it with that of a population of early endosomes. Both endosomes and lysosomes exhibited ATP-dependent acidification, but proton influx rates were 2.4- to 2.7-fold greater for endosomes than for lysosomes because of differences in both buffering capacity and acidification rates, suggesting that endosomes exhibited greater numbers or rates of proton pumps. Lysosomes, however, exhibited a more acidic steady-state pHi due in part to a slower proton leak rate. Changes in medium Cl- increased acidification rates of endosomes more than lysosomes, and the lysosome ATP-dependent interior-positive membrane potential was only partially eliminated by high-Cl- medium. Permeability studies suggested that lysosomes were less permeable to Na+, Li+, and Cl- and more permeable to K+ and PO4(2-) than endosomes. Na-K-adenosine-triphosphatase did not appear to regulate acidification of either vesicle type. Endosome and lysosome acidification displayed similar inhibition profiles to N-ethylmaleimide, dicyclohexyl-carbodiimide, and vanadate, although lysosomes were somewhat more sensitive [concentration producing 50% maximal inhibition (IC50) 1 nM] to bafilomycin A1 than endosomes (IC50 7.6 nM). Oligomycin (1.5-3 microM) stimulated lysosome acidification due to shunting of membrane potential. Overall, acidification of endosomes and lysosomes was qualitatively similar but quantitatively somewhat different, possibly related to differences in the density or rate of proton pumps as well as vesicle permeability to protons, anions, and other cations. PMID- 8238316 TI - Structural determinants of substrate specificity of the erythrocyte anion transporter. AB - The human erythrocyte anion transport protein (AE1) mediates the rapid, tightly coupled, electroneutral transmembrane exchange of bicarbonate for chloride. AE1 transports a wide range of oxyanions, such as phosphate, sulfate and the physiological substrate bicarbonate. In this study, the transport characteristics of the selenium based oxyanions selenite (SeO3(2-)) and selenate (SeO4(2-)) were determined. The pH dependence of selenate influx was consistent with a titratable carrier having a extracellular pK value of 5.67 +2- 0.09. In contrast, the pH dependence of selenite influx had a maximum near pH 7.0, consistent with a hypothesis proposed by Labotka and Omachi (J. Biol. Chem. 263: 1166-1173, 1988) that the pH maximum of the transport of titratable anions is located at the midpoint between the pK of the carrier (5.7) and the pK of the titratable anion (8.3). Analysis of the transport rates and structures of these as well as a variety of other oxyanions reported in the literature suggested that oxyanions bind in a three-oxygen atom binding site, and that the formation of the transition state necessary for transport is sterically restrained by oxyanions that protrude in a direction perpendicular to the three-oxygen binding plane. This hypothesis can be used to predict the relationship between the transport rates of many oxyanions reported in the literature and should prove useful in helping to understand the molecular mechanism of AE1 mediated transport. PMID- 8238317 TI - Metabolism of a long-chain diacylglycerol by permeabilized A10 smooth muscle cells. AB - The regulatory effects of diacylglycerol (DAG) second messengers will be terminated by metabolism. A long-chain DAG, 1-palmitoyl-2-[1-14C]oleoyl-sn glycerol (2-[14C]POG), was metabolized by cultured A10 smooth muscle cells after permeabilization by preincubation with 340 U/ml alpha-toxin from Staphylococcus aureus. In contrast to results with the cell-permeable DAG analogue, dioctanoyl glycerol ([3H]diC8), no appreciable 2-[14C]POG degradation could be detected in control A10 cells not treated with alpha-toxin. With permeabilized A10 cells, 2 [14C]POG was mainly converted into lipolytic products of a lipase pathway, monoacylglycerol (MG) and fatty acid (FA); very little radioactivity was incorporated into triacylglycerol (TG) or phospholipid (PL) via reactions catalyzed by either DAG acyltransferase, cholinephosphotransferase, or DAG kinase. Similar results were obtained in experiments with 1-stearoyl-2-[1 14C]arachidonoyl-sn-glycerol. The conversion of 2-[14C]POG into PL and TG was not enhanced by the addition of 1 mM ATP-MgCl2, 1 mM CDP-choline, or 1 mM oleoyl-CoA to the alpha-toxin-treated A10 cells. The formation of FA and MG by permeabilized A10 cells was inhibited by DAG lipase inhibitors, U-57,908 (50 microM) and tetrahydrolipstatin (1-25 nM). The predominant contribution of the lipase pathway to the metabolism of a long-chain DAG, 2-[14C]POG, by alpha-toxin-treated A10 cells is similar to results for the degradation of [3H]diC8 by intact A10 cells. PMID- 8238318 TI - Adenosine A1 receptors and erythropoietin production. AB - N6-cyclopentyladenosine (CPA), a selective adenosine A1 receptor agonist, in a concentration range of 10(-9) to 10(-7) M, produced a significant decrease in erythropoietin (EPO) levels in a human hepatocellular carcinoma (Hep G2) cell culture (medium levels of EPO, 91.81 +/- 1.61 and 94.36 +/- 0.97% of control, respectively) after 24 h incubation in a hypoxic atmosphere. CPA, at a concentration of 10(-9) M, also produced a significant decrease in Hep G2 cell levels of adenosine 3',5'-cyclic monophosphate (cAMP; 78.13 +/- 3.89% of control) after 2 h incubation. CPA (10(-9) M) also significantly inhibited forskolin stimulated increases in EPO production and cAMP accumulation in Hep G2 cells. On the other hand, 2-[p-(2-carboxyethyl)phenethyl-amino]-5'-N ethylcarboxamidoadenosine (CGS-21680), a selective adenosine A2-receptor agonist, produced no significant change in EPO production in a dose range of 10(-10) to 10(-6) M but increased cAMP accumulation at 10(-6) M. A1-receptor binding assays using N6-[3H]cyclohexyladenosine revealed a single type of adenosine receptor binding site on Hep G2 cell membranes with a dissociation constant of 71.4 nM and a binding capacity of 1,530 fmol/mg protein. These results indicate that Hep G2 cells contain high-affinity adenosine A1 receptors that are linked to decreased cAMP accumulation and EPO production. PMID- 8238319 TI - AVP-induced activation of MAP kinase in vascular smooth muscle cells is mediated through protein kinase C. AB - Arginine vasopressin (AVP) has been shown to stimulate tyrosine phosphorylation and activation of p42 mitogen-activated protein (MAP) kinase (p42MAPK) in vascular smooth muscle cells (VSMC). In VSMC, AVP increases free intracellular Ca2+ concentration ([Ca2+]i) and activates protein kinase C (PKC) through activation of phospholipase C. The contribution of PKC and [Ca2+]i in p42MAPK regulation was therefore determined. Activation of PKC by phorbol 12-myristate 13 acetate (PMA) stimulated tyrosine phosphorylation and activation of p42MAPK to the same extent as AVP. Inhibition of PKC by staurosporine or downregulation of PKC by PMA pretreatment abolished AVP-induced stimulation of p42MAPK. When [Ca2+]i was elevated to the same level as with AVP, using either ionomycin (0.1 microM) or thapsigargin (0.1 microM), MAP kinase was only partially activated. Elevation of [Ca2+]i to supraphysiological levels by 1 microM ionomycin stimulated MAP kinase activity to the same extent as AVP. This effect was blocked by downregulation of PKC. The intracellular Ca2+ chelator BAPTA [1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid] blocked AVP-induced [Ca2+]i increase but did not affect AVP stimulation of p42MAPK. Thus AVP-induced activation of p42MAPK requires only the activation of PKC but not an increase in [Ca2+]i. PMID- 8238320 TI - G protein-mediated inhibition of inwardly rectifying K+ channels in guinea pig chromaffin cells. AB - Properties of inwardly directed rectification and its G protein-mediated inhibition in guinea pig chromaffin cells were studied using the whole cell version of the patch-clamp technique. The current-voltage (I-V) relationship for plateau currents in response to a 50-ms pulse showed an inwardly directed rectification between -80 and -140 mV and a negative slope at more negative potentials in normal solution. Replacement of Na+ with N-methyl-D-glucamine (NMDG) in the perfusate did not alter the plateau I-V relationship between -110 and -130 mV but did abolish the negative slope below -140 mV. The zero current or resting membrane potential in the NMDG solution was in fair agreement with the equilibrium potential for K+. The chord conductance-voltage relationship showed a good fit with the Boltzmann equation and shifted along the voltage axis by an approximate change in driving force on K+ when K+ concentration was increased. External Cs+ and Ba2+ produced a voltage-dependent inhibition of the inwardly directed rectification. These results indicate that inwardly rectifying (IR) K+ channels are mediating an inwardly directed rectification. Intracellular dialysis with guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) produced a complete suppression of this IR K+ channel, irrespective of treatment with pertussis toxin. Adding GTP or guanosine 5'-O-(2-thiodiphosphate) to the patch solution resulted in a decrease in GTP gamma S inhibition of the K+ current. Internal application of vanadate was without effect. Time course of the inhibition of the IR K+ current coincided in part with that of inactivation of a nonselective cation current. In conclusion, IR K+ channels in the chromaffin cell are subject to G protein-mediated inhibition. PMID- 8238321 TI - ATP-inhibitable Cl- channel in apical membranes of cultured rabbit cortical collecting duct cells. AB - Excised patches of apical membranes from immunodissected rabbit cortical collecting duct cells in primary culture were studied by the patch-clamp technique. Barium (1 mM) and tetraethylammonium chloride (5 mM) were added to all solutions to block potassium channel activity. A unique channel was observed that exhibited inward rectification under symmetrical ionic conditions with a measured chord conductance of 54.0 +/- 2.5 pS at -80 mV (n = 11) and 22.1 +/- 1.7 pS at +80 mV (n = 5). This channel was chloride selective, with a PNa:PCl of 0.16 (n = 3). Kinetic analysis revealed a voltage-independent open-time probability of 0.80 +/- 0.07 (n = 6). Open-time probability within bursts was 0.96 +/- 0.01. Addition of ATP to the cytosolic surface of the channel resulted in a dose-dependent decrease in open probability, with a threshold effect at 10(-4) M, due to a reduction in burst open time. The effect of ATP was immediate, rapidly reversible at room temperature, and mimicked by GTP, adenosine 5'-O-(3-thiotriphosphate), and guanosine 5'-O-(3-thiotriphosphate). This channel may link epithelial chloride permeability to cellular ATP content in the rabbit cortical collecting duct. PMID- 8238322 TI - Intracellular Ca2+ signaling induced by vasopressin, ATP, and epidermal growth factor in epithelial LLC-PK1 cells. AB - The free calcium concentrations in nucleus ([Ca2+]n) and in cytoplasm ([Ca2+]c) of cultured renal LLC-PK1 epithelial cells were estimated by confocal laser microscopy. No difference between the resting mean [Ca2+]n and [Ca2+]c was found. During stimulation with maximal effective concentrations of arginine vasopressin (AVP) or the purinergic agonist ATP, the transient Ca2+ rise was followed mostly by a decline to basal levels. A differential rise could be observed when the increase in [Ca2+]n attained higher values than [Ca2+]c. In 50-60% of the cells, epidermal growth factor (EGF) also induced a transient Ca2+ rise, and a differential increase ([Ca2+]n > [Ca2+]c) was found. The G protein-linked stimuli AVP and ATP were however quantitatively much more efficacious at stimulating the [Ca2+]n and [Ca2+]c increases than was EGF. To investigate whether AVP, ATP, and EGF released Ca2+ from distinct or overlapping stores, the agonists were sequentially added. AVP and ATP applied after EGF in Ca(2+)-free medium elicited an increase in [Ca2+]n and [Ca2+]c that was not significantly lower than the release of Ca2+ in control cells without EGF prestimulation. Similarly, the amplitude of the Ca2+ responses attained by EGF in cells prestimulated by ATP or AVP was comparable to the response in naive cells. Neither EGF, ATP, nor AVP evoked a Ca2+ signal after thapsigargin treatment, indicating that the intracellular Ca2+ pools stimulated by all these agonists are part of the thapsigargin-sensitive Ca2+ pools. In contrast, when ATP was applied after AVP in Ca(2+)-containing as well as in Ca(2+)-free solutions, the Ca2+ transients were lower as compared with the response without preincubation. No differential rise could be found in Ca(2+)-free conditions. An explanation could be the use of different phospholipase C isozymes by the different receptor types, which possibly gives rise to the mobilization of different Ca2+ pools. PMID- 8238323 TI - Calcium-activated K+ channels as modulators of human myometrial contractile activity. AB - The role of Ca(2+)-activated potassium (KCa) channels in the regulation of membrane potential, intracellular free calcium ([Ca2+]i) and contraction was investigated in uterine smooth muscle and myometrial cells. In an immortalized human myometrial cell line, oxytocin increased [Ca2+]i and [3H]inositol phosphate formation. Relaxin attenuated the oxytocin-induced increase in [Ca2+]i. In cell attached patches, membrane depolarization activated a large-conductance KCa channel (179 +/- 4 pS). Iberiotoxin (IbTX), a potent blocker of "maxi" KCa channels (A. Galvez, G. Gimenez-Gallego, J. P. Reuben, L. Roy-Contanciin, P. Feigenbaum, G. J. Kaczorowski, and M. L. Garcia. J. Biol. Chem. 265: 11083-11090, 1990) produced long closed events (approximately 6 min) in these channels. In agreement with this blockage, IbTX depolarized the cells by 9.8 +/- 2.8 mV and caused a dose-dependent increase in [Ca2+]i with a half-maximal effective concentration of 0.79 nM. IbTX also caused phasic contractions in human myometrial strips and increased both the frequency and force of spontaneous contractions in estrogen-primed rat myometrial strips. Moreover, myometrial contractility was also affected by 1 mM tetraethylammonium, a concentration that blocks uterine smooth muscle KCa channels when applied to the extracellular side (G. J. Perez, L. Toro, S. D. Erulkar, and E. Stefani. Am. J. Obstet. Gynecol. 168: 652-660, 1993). These results strongly suggest that the large conductance KCa channels may actively participate in the control of human myometrial cell membrane potential and [Ca2+]. PMID- 8238324 TI - Fluid-phase endocytosis by in situ cardiac myocytes of rat atria. AB - Fluid-phase endocytosis (FPE) associated with recycling of fused plasmalemma secretory granules or membranes and/or membrane receptors by in situ cardiac myocytes was studied at 37 degrees C in vitro noncontracting adult rat atrial preparations. Measurements included 1) the volume (VS*) of the compartment consisting of presumptive endocytotic vesicles and the endosomes or lysosomes transiently in continuity with them (S*), which internalizes [14C]-sucrose but is inaccessible to simultaneously measured [methoxy-3H]inulin, 2) the kinetics of [14C]sucrose efflux from S*, and 3) morphometry to quantify interstitial space and non-heart muscle cells. Vs* (0.39 +/- 0.04 ml/g dry atrium for unstretched atria at 37 degrees C) was 1) variable over a 3.7-fold range under various experimental conditions, 2) significantly increased by neomycin or by lowering the temperature to 18 degrees C, and 3) significantly decreased by alpha 1 adrenergic stimulation. Analysis of sucrose efflux kinetics confirmed the presence of an intramyocytic sucrose-containing compartment. A smaller inulin inaccessible sucrose space (S*) was also present in right ventricle. Thus, during FPE, vesicles and endosomes initially containing high (extracellular) Ca2+ and Cl concentrations continually enter, circulate within, and undergo exocytosis from myocardial cells. PMID- 8238325 TI - Ovine fetal protein metabolism during decreased glucose delivery. AB - During decreased fetal glucose delivery, endogenous substrates increasingly support metabolic rate. If oxidation of fetal amino acid stores (i.e., protein) is to provide such substrate, either protein synthesis must decrease or protein breakdown must increase. Both of these changes would diminish fetal protein accretion, an important component of fetal growth. This study was performed, therefore, to find if decreased glucose delivery alters fetal leucine metabolism. Catheters were placed in six sheep fetuses under maternal general anesthesia, and, after 6 days recovery, fetal leucine metabolism was measured by infusion of [1-14C]leucine before and after fetal glucose delivery was decreased by infusing insulin to the ewe. Later (2 days) the experiment was repeated in reverse order. Decreased fetal glucose delivery was associated with a 19% decrease in the rate of fetal leucine disposal (P < 0.001), a 42% decrease in the rate of exogenous leucine uptake (P < 0.01), and no change in the rate of fetal leucine decarboxylation. The use of leucine for protein synthesis by the fetus fell by 23% (P < 0.001), whereas the rate of protein breakdown did not change. Consequently, during decreased fetal glucose delivery, leucine accretion into fetal proteins was 28% (P < 0.02) of the control rate. In summary, fetal oxidation of amino acids derived from increased protein breakdown is not the source of endogenous substrate needed by the fetus with restricted glucose availability. Fetal protein synthesis did decrease, however, diminishing the accretion of leucine into protein. PMID- 8238326 TI - Dietary arginine uptake by the splanchnic region in adult humans. AB - To determine the uptake of dietary arginine and leucine by the splanchnic region, two experiments were carried out, each involving four healthy young adult men who received a diet supplying 1 g protein.kg-1.day-1 for 7 and 10 days before conducting a primed constant tracer infusion protocol. In study 1, subjects received for 8 h (3-h fast; 5-h fed state, achieved by a constant intragastric infusion of the diet formula) L-[5,5-2H2; guanidino-15N2]arginine ([M4]Arg), L [guanidino-13C]arginine ([13C]Arg), and L-[5,5,5-2H3]leucine ([2H3]Leu) simultaneously by an intragastric infusion on day 7 and a repeat of this protocol on day 10 except with tracer administration given by vein. Plasma arginine fluxes were essentially the same for the two arginine tracers but differed significantly with route of administration. In study 2 the subjects received on day 7 a constant intravenous infusion of [13C]Arg and [2H3]Leu and a simultaneous intragastric infusion of [M4]Arg and [1-13C]leucine. On day 10 the routes of administration of these tracer pairs were reversed. During the fed state in study 1, splanchnic uptake of dietary arginine was 31 +/- 10 and 34 +/- 8%, based on the [13C]Arg and [M4]Arg tracers, respectively, and it was significantly higher (P < 0.01) than for leucine, which was 10 +/- 6%. In study 2, splanchnic uptake of dietary arginine, estimated from a series of tracer-protocol combinations for the fed state, was approximately 38% compared with a lower (P < 0.01) value of approximately 15% for leucine. PMID- 8238327 TI - Leucine metabolism at graded amino acid intakes in children receiving parenteral nutrition. AB - To assess the response of protein turnover to graded levels of amino acid (AA) intakes, leucine kinetics were determined in six 8- to 16-yr-old patients in a stable nutritional status receiving home parenteral nutrition (PN) for short bowel syndrome or intestinal pseudo-obstruction syndrome. Although daily energy intake was kept constant at 68.7 +/- 13 kcal/kg lean body mass (LBM) with 25.4 +/ 3.6% lipid, patients were given, for three consecutive 7-day periods, 0.7, 1.5, or 2.5 g AA.kg LBM-1.day-1, with the order of the regimens being randomized. On day 7 of each period, a 4-h infusion of L-[1-13C]leucine was performed during intravenous feeding; plasma [13C]ketoisocaproate and expired 13CO2 enrichments were used to assess whole body leucine turnover (Ra), oxidation rate (Ox), nonoxidative disposal [an estimate of protein synthesis (S)], and leucine derived from protein breakdown (B). Urine collection (24 h) was performed for determination of nitrogen excretion. Results indicate a dose-dependent rise in plasma leucine concentration, Ra, and Ox but no significant change in B. There was a significant increase of S (P = 0.04 analysis of variance) with increased AA intakes as well as net leucine balance (P = 0.02). Results are consistent with improved leucine balance, when leucine intake increases, despite increased leucine oxidation. The net protein gain observed with higher AA intakes may suggest a beneficial effect for children receiving long-term PN. PMID- 8238328 TI - Metabolic acidosis inhibits growth hormone secretion in rats: mechanism of growth retardation. AB - To test the hypothesis that growth retardation in nonanion gap acidosis may be associated with impairment of growth hormone (GH) secretory patterns, we examined GH secretion in rats made acidotic with ammonium chloride ingestion. Considerable growth retardation was demonstrated in pair-fed and acidotic rats after 1 wk of ammonium chloride ingestion compared with control. With stable metabolic acidosis sustained on the 8th day of experiment, pulsatile secretion of GH was evaluated by blood samples drawn every 10 min for 6 h. Using deconvolution analysis to quantitate in vivo GH secretory rates, we found significant inhibition of pulsatile GH secretion in acidotic rats. Changes in amplitude of GH pulses and mean mass of GH pulses correlated with changes in body weight. These studies showed that chronic metabolic acidosis causes growth impairment, reduced food efficiency, and amplitude-specific inhibition of pulsatile secretion of GH. We propose that this GH axis suppression, whether mediated by decreased nutrients or not, contributes significantly to growth failure in children with renal tubular acidosis. Because a similar degree of inhibition of GH secretion was seen in pair fed rats, we infer that insufficient calorie intake in metabolic acidosis may contribute to disruption of normal GH secretion patterns. These changes in GH secretion were specific, because acidotic rats were different from pair-fed controls in that they showed no change in either half-life of GH in circulation or in pulse frequency. Such observations offer a rationale for more detailed clinical investigations into the impact of metabolic acidosis on physiological regulation of pulsatile GH secretion in humans. PMID- 8238329 TI - Effect of age, calcium source, and radiolabeling method on whole body 47Ca retention in the rat. AB - In a longitudinal study we determined the effect of animal age as well as Ca source and radiolabeling method on Ca bioavailability by measuring whole body 47Ca retention (WBR) in male Sprague-Dawley rats. The WBR assay was performed without surgery or anesthesia, and the same groups of animals were studied at 8, 16, 20, and 32 wk of age. Rats were administered a 6-mg radiolabeled oral dose of Ca as Ca citrate malate (CCM) or intrinsically or extrinsically labeled CaCO3 or hydroxyapatite (HAP). Fractional Ca retention was measured from the 72-h postdose WBR divided by WBR at time 0. WBR was significantly affected by Ca source with CCM > CaCO3 > HAP at all ages (P < 0.001). The rank order and relative bioavailabilities of these Ca salts in the rat model agreed well with literature values for Ca absorption in adult humans. Although percent WBR decreased significantly with advancing age (P < 0.001), the mean rate of decline (-3.4%/wk) was not affected by Ca source. Extrinsic radiolabeling overestimated (approximately 20%) Ca bioavailability when the rats were young. However, the magnitude of this effect diminished with advancing animal age and was not significant across all ages (repeated measures analysis of variance P = 0.10). PMID- 8238330 TI - Adaptive changes in zonation for gluconeogenic capacity in liver lobules of cold exposed rats. AB - The rate of gluconeogenesis from lactate increased in perfused livers after exposure of rats to cold for 5 days, and it returned to the control rate after 20 days [M. Shiota, T. Tanaka, and T. Sugano. Am. J. Physiol. 249 (Endocrinol. Metab. 12): E281-E286, 1985.]. The relationship between the increased gluconeogenic activity and its zonal distribution in liver lobules was studied in cold-exposed rats that had been starved for 24 h by examination of preparations enriched for periportal hepatocytes (PP-H) and for perivenous hepatocytes (PV-H), which had been isolated by the digitonin-collagenase perfusion technique. In the control group, the rate of gluconeogenesis from lactate or alanine was three times higher in PP-H than in PV-H. The rate of gluconeogenesis from these substrates in PP-H was not changed by exposure of rats to cold. The rates of PV-H increased to the level in PP-H after 5 days of exposure of rats to cold and then returned to the control rates after 20 days. The rate of gluconeogenesis from fructose was not altered in either preparation of cells by cold treatment of rats. The change in gluconeogenic capacity in PV-H caused by exposure of rats to cold was unrelated to changes in the activity of the malate-aspartate shuttle and of pyruvate kinase. The increased capacity in mitochondrial respiration was observed in both preparations of cells by cold treatment of rats for 5 days. The activity of phosphoenolpyruvate carboxykinase was higher in PP-H than in PV-H in the control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238331 TI - Long-term effects on pancreatic function of feeding a HC formula to rats during the preweaning period. AB - We have investigated pancreatic changes associated with hyperinsulinemia and an insulin secretory response to an oral glucose load in a new rat model for obesity. Male Sprague-Dawley pups were reared on a high-carbohydrate (HC) or high fat (HF) formula by gastrostomy during the suckling period and were weaned onto a stock diet. These animals remained either nutritionally unchallenged or challenged with a high-sucrose diet during the postweaning period. The HC formula fed animals showed increased insulin concentrations in the plasma and pancreas and also showed impaired insulin secretory response compared with mother-fed control or HF animals in adult life. Immunocytochemical and morphometric studies revealed that hyperinsulinemia in the HC animals during the preweaning period and also in adult life was associated with hypertrophy of beta-cells in the pancreas. The results show that consumption of a HC formula during the suckling period influences pancreatic islet morphology resulting in hyperinsulinemia which eventually leads to the development of obesity later in adult life. PMID- 8238332 TI - Regulation of maternal IGF-I by placental GH in normal and abnormal human pregnancies. AB - Throughout gestation, maternal insulin-like growth factor I (IGF-I) increases progressively despite suppressed pituitary growth hormone (GH) secretion. We have previously shown that in normal pregnancy, a specific placental GH variant, rather than human placental lactogen (hPL), substitutes for pituitary GH in the regulation of maternal IGF-I. We studied the maternal IGF-I secretion in a cohort of 286 normal and abnormal pregnancies (617 blood samples). Regardless of pathology and gestational age, IGF-I values correlated with corresponding placental GH but not with hPL values. Similar correlations were evidenced for each 2-wk gestational period between 32 and 39 wk. In pathological pregnancies, when only those hormonal results that are obtained before any treatment are considered and diabetes is excluded, IGF-I levels were closely related to corresponding placental GH, but not to hPL. In women with a fetoplacental unit disorder, low placental GH levels resulted in low IGF-I and in a secondary pituitary GH increase, whereas in patients without detectable impairment of the fetoplacental unit normal placental GH corresponded to normal IGF-I. These results suggest that in pathological as well as in normal pregnancy, placental GH, and not hPL, substitutes for pituitary GH to regulate the maternal IGF-I secretion. PMID- 8238333 TI - Hypothalamic and gastric myoelectrical responses during vection-induced nausea in healthy Chinese subjects. AB - The physiology of nausea, a uniquely human symptom, is poorly understood. The purpose of this study was to measure the temporal sequences of neurohormonal responses and gastric myoelectrical activity in healthy subjects during the rotation of an optokinetic drum that produced nausea and other symptoms of motion sickness. Plasma catecholamines, vasopressin, and cortisol were measured at baseline, during minutes 1-5, 6-10, and 11-15 of drum rotation, and after rotation stopped. Electrogastrograms were recorded throughout the study. Twelve subjects (80%) developed nausea and 4-9 cycles/min of gastric tachyarrhythmias; three subjects had no nausea and no gastric dysrhythmias. Tachyarrhythmias began 3.4 +/- 0.8 min after the onset of drum rotation, and nausea was reported, on average, 3 min later. During minutes 6-10 of drum rotation, vasopressin levels significantly increased in the subjects with nausea compared with subjects without nausea (P < 0.04). In the subjects with nausea, epinephrine and vasopressin increased significantly (P < 0.05) compared with baseline during minutes 6-10 and 11-15 of drum rotation. As nausea resolved during recovery, vasopressin decreased by 74%, whereas epinephrine increased 13%. We conclude that 1) in nauseated subjects, endogenous vasopressinergic and sympathetic circuits are activated before hypothalamic-pituitary-adrenal pathways, 2) plasma vasopressin levels correlate most closely with the temporal onset and resolution of nausea, and 3) peripheral gastric dysrhythmias may have a role in activating central vasopressinergic neurons. PMID- 8238334 TI - Importance of brain IL-1 type II receptors in fever and thermogenesis in the rat. AB - Interleukin-1 (IL-1) acts centrally to induce fever and thermogenesis in rodents. The central actions of IL-1 alpha and IL-1 beta apparently involve different mechanisms, and the effects of IL-1 beta are not consistent with interaction with a type I (IL-1RI) 80-kDa receptor. In the present study the involvement of the type II IL-1 receptor (IL-1RII) was tested in the rat by examining the effects of central injection of a monoclonal antibody (ALVA-42), which blocks the IL-1RII. Pretreatment of rats with ALVA-42 (6 micrograms icv) inhibited the thermogenic and pyrogenic responses to intracerebroventricular injection of 5 ng (but not 50 ng) of IL-1 beta in conscious rats but did not significantly modify responses to IL-1 alpha. ALVA-42 also failed to modify the responses to peripherally administered IL-1 beta (1 microgram) but significantly attenuated the pyrogenic and thermogenic responses to peripheral (125 micrograms) or central (1 microgram) injection of endotoxin. These data indicate that IL-1RII mediates the central effects of a low dose of IL-1 beta, but not IL-1 alpha, on fever and thermogenesis in the rat. They also imply that responses to endotoxin are due, at least in part, to the activation of IL-1RII by IL-1 beta released within the brain and that effects of peripherally injected IL-1 beta involve different mechanisms, probably associated with IL-1RI. PMID- 8238335 TI - Effects of blockade of fatty acid oxidation on whole body and tissue-specific glucose metabolism in rats. AB - We examined the effect of the long-chain fatty acid oxidation blocker methyl palmoxirate (methyl 2-tetradecyloxiranecarboxylate, McN-3716) on glucose metabolism in conscious rats. Fasted animals [5 h with or without hyperinsulinemia (100 mU/l) and 24 h] received methyl palmoxirate (30 or 100 mg/kg body wt po) or vehicle 30 min before a euglycemic glucose clamp. Whole body and tissue-specific glucose metabolism were calculated from 2-deoxy-[3H]-glucose kinetics and accumulation. Oxidative metabolism was assessed by respiratory gas exchange in 24-h fasted animals. Pyruvate dehydrogenase complex activation was determined in selected tissues. Methyl palmoxirate suppressed whole body lipid oxidation by 40-50% in 24-h fasted animals, whereas carbohydrate oxidation was stimulated 8- to 10-fold. Whole body glucose utilization was not significantly affected by methyl palmoxirate under any conditions; hepatic glucose output was suppressed only in the predominantly gluconeogenic 24-h fasted animals. Methyl palmoxirate stimulated glucose uptake in heart in 24-h fasted animals [15 +/- 5 vs. 220 +/- 28 (SE) mumol x 100 g-1 x min-1], with smaller effects in 5-h fasted animals with or without hyperinsulinemia. Methyl palmoxirate induced significant activation of pyruvate dehydrogenase in heart in the basal state, but not during hyperinsulinemia. In skeletal muscles, methyl palmoxirate suppressed glucose utilization in the basal state but had no effect during hyperinsulinemia; pyruvate dehydrogenase activation in skeletal muscle was not affected by methyl palmoxirate under any conditions. The responses in skeletal muscle are consistent with the operation of a mechanism similar to the Pasteur effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238336 TI - Characterization of endothelin receptors in the anterior pituitary gland. AB - To characterize endothelin (ET) receptors modulating pituitary hormone secretion, potencies of ET-like agonists were compared on prolactin (PRL), thyrotropin (TSH), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) secretion from primary cultures of female rat pituitary cells. ET-1 was more potent than ET 3 in all cases. Sarafotoxin (SRTX) S6b an ETA agonist, was also more potent than ET-3 in all cases. SRTX-c, an ETB receptor agonist, was inactive. The ET-1-to-ET 3 potency ratio was three orders of magnitude higher on PRL or TSH secretion than on LH and FSH secretion, whereas SRTX-b-to-ET-3 potency ratios were similar on all four hormones. The ETA antagonist BQ-123 caused a parallel dextral displacement of dose-response curves of ET-1 and ET-3 on all four hormones. Schild regressions for BQ-123 on ET-1-induced PRL, TSH, LH, and FSH secretion indicated that BQ-123 has a similar affinity for the receptors mediating ET-1's effects. When BQ-123 was assessed against ET-3, Schild regressions indicated greater affinity for ET-3 on lactotrophs and thyrotrophs than gonadotrophs. Thus changes in pituitary hormone secretion are mediated by ETA-like receptors. ET receptors in lactotrophs and thyrotrophs are clearly distinguishable from gonadotrophs. We suggest the existence of distinct ETA receptor subtypes (ETA1 and ETA2) on these differing pituitary cells. PMID- 8238337 TI - Effects of insulin on the cardiovascular integrating mechanisms of brain stem in cats. AB - In 65 cats anesthetized with alpha-chloralose and urethane, the effects of insulin on cardiovascular responses to stimulation of various structures in the brain stem were studied. The threshold dose of insulin injected intravenously that produced systemic hypoglycemia was 5-10 U/kg. Subthreshold hypoglycemic doses of insulin were used intracerebroventricularly (0.25 U/kg) or intracerebrally (2 mU in 200 nl). Sixty minutes after intravenous insulin, when serum glucose concentrations decreased from 158 to 43 mg/100 ml, pressor responses to stimulation of the periaqueductal gray of midbrain (PAG), locus coeruleus (LC), dorsal medulla (DM), ventrolateral medulla (VLM), and parvocellular reticular nucleus (PVC) decreased significantly. Depressor and bradycardiac response to stimulation of paramedian reticular nucleus or dorsal motor nucleus of vagus (DMV) decreased significantly as well. Thirty minutes after intracerebroventricular insulin, pressor responses of PAG, DM, and the bradycardiac response of DMV decreased significantly. Thirty minutes after intracerebral insulin, pressor responses and renal nerve activities of LC (but not PAG), VLM, DM, and PVC decreased significantly. A similar but faster onset (5 min) of depression of cardiovascular responses on stimulating the LC, VLM, DM, and PVC was observed in another six acutely midcollicular-decerebrate cats recovered from halothane anesthesia. These findings suggest that insulin directly inhibits the vasomotor structures of the brain stem and decreases the pressor responses to stimulation. PMID- 8238338 TI - Creatine kinase isoenzymes are highly regulated during pregnancy in rat uterus and placenta. AB - Creatine kinase (CK) isoenzymes play a central role in energy transfer. Expression of CK isoenzymes in rat uterus and placenta was analyzed to evaluate their contribution to energy metabolism during pregnancy and delivery. Tissue from the uterine horns and placentas of pregnant rats from day 14 of gestation to 17 days postpartum was analyzed for expression of "brain" CK (BCK) and ubiquitous mitochondrial CK (uMtCK) mRNA, protein, and enzyme activity. uMtCK mRNA expression is high in prepartum uterus, but rapidly falls (> 10-fold) after delivery to a nadir at 7 days postpartum. Prepartum BCK mRNA expression is coordinate with uMtCK but has a 15-fold greater expression than uMtCK. Both CK mRNAs rise by 17 days postpartum. Both BCK and uMtCK mRNA expressions are strongly induced in placenta at 20 days gestation with a rapid fall (> 6-fold) immediately before delivery. Protein expression of BCK and uMtCK is also coordinate. However, analysis of mRNA and protein expression indicates that significant posttranscriptional regulation of both kinds of CK also occurs. CK activity in uterus and placenta reflects BCK expression. By immunohistochemistry, BCK and uMtCK protein expression is highly localized in the placenta and endometrium of prepartum uterus. This expression shifts entirely to the uterine smooth muscle by 17 days postpartum. uMtCK mRNA expression is rapidly induced by beta-estradiol in vitro (> 6-fold), demonstrating estrogen-responsive elements in the uMtCK nuclear gene. Thus a second isoenzyme of CK, uMtCK, is expressed in rat uterus and placenta and is highly regulated with BCK. These results suggest an important role for CK in the maintenance and termination of pregnancy. PMID- 8238339 TI - 14C-labeled propionate metabolism in vivo and estimates of hepatic gluconeogenesis relative to Krebs cycle flux. AB - Purposes of this study were 1) to estimate in humans, using 14C-labeled propionate, the rate of hepatic gluconeogenesis relative to the rate of Krebs cycle flux; 2) to compare those rates with estimates previously made using [3 14C]lactate and [2-14C]acetate; 3) to determine if the amount of ATP required for that rate of gluconeogenesis could be generated in liver, calculated from that rate of Krebs cycle flux and splanchnic balance measurements, previously made, and 4) to test whether hepatic succinyl-CoA is channeled during its metabolism through the Krebs cycle. [2-14C]propionate, [3-14C]-propionate, and [2,3 14C]succinate were given along with phenyl acetate to normal subjects, fasted 60 h. Distributions of 14C were determined in the carbons of blood glucose and of glutamate from excreted phenylacetylglutamine. Corrections to the distributions for 14CO2 fixation were made from the specific activities of urinary urea and the specific activities in glucose, glutamate, and urea previously found on administering [14C]-bicarbonate. Uncertainties in the corrections and in the contributions of pyruvate and Cori cyclings limit the quantitations. The rate of gluconeogenesis appears to be two or more times the rate of Krebs cycle flux and pyruvate's decarboxylation to acetyl-CoA, metabolized in the cycle, less than one twenty-fifth the rate of its decarboxylation. Such estimates were previously made using [3-14C]lactate. The findings support the use of phenyl acetate to sample hepatic alpha-ketoglutarate. Ratios of specific activities of glucose to glutamate and glucose to urinary urea and expired CO2 indicate succinate's extensive metabolism when presented in trace amounts to liver. Utilizations of the labeled compounds by liver relative to other tissues were in the order succinate = lactate > propionate > acetate. ATP required for gluconeogenesis and urea formation was approximately 40% of the amount of ATP generated in liver. There was no channeling of succinyl-CoA in the Krebs cycle in the hepatic mitochondria. PMID- 8238340 TI - Evidence that intrarenal bradykinin plays a role in regulation of renal function. AB - Bradykinin (BK) is produced by the kidney, but the role of the renal kallikrein kinin system (KKS) in the control of renal function is not understood. We studied the effects of intrarenal infusion of the BK antagonist, D-Arg-Arg-Pro-Hyp-Gly Thi-Ser-D-Phe-Thi-Arg-trifluoroacetic acid (BKA, n = 5) and BK (n = 4) alone or combined with antagonist (BKA 0.025 ng.kg-1 x min-1 + BK 0.25 ng.kg-1 x min-1, n = 4) in uninephrectomized conscious dogs in sodium balance at 10 and 80 meq/day. During low sodium intake, administration of BKA (infusions from 0.025 to 2.5 ng.kg-1 x min-1) caused a significant antidiuresis (P < 0.0001) and antinatriuresis (P < 0.0001) and a decrease in fractional sodium excretion (P < 0.0001). There were no changes in estimated renal plasma flow (RPF) or glomerular filtration rate during intrarenal administration of BKA at 0.025 and 0.25 ng.kg-1 x min-1. A dose of 2.5 ng.kg-1 x min-1 BKA caused a significant decrease in RPF. There were no changes in plasma aldosterone concentration, plasma renin activity, or systemic arterial pressure during intrarenal BKA administration. At 80 meq/day sodium balance (n = 5), intrarenal administration of BKA did not cause any systemic or renal effects. Intrarenal administration of BK at 0.25 ng.kg-1 x min 1 during low sodium balance caused an increase in urine flow rate and urinary sodium excretion. Coinfusion of BK with BKA completely abrogated the renal excretory changes induced by BKA. These data suggest that intrarenal KKS plays a role in control of renal function largely by a tubular mechanism during low sodium intake. PMID- 8238341 TI - Circulating insulin levels are related to bone density in normal postmenopausal women. AB - We recently established that the dependence of bone mineral density (BMD) on body weight in women is mainly attributable to a close relationship between total body fat mass and BMD. The present study assesses whether this latter relationship might be contributed to by the hormones insulin or amylin, both of which may influence fat mass and calcium metabolism. Fifty-three normal postmenopausal women underwent a 75-g glucose tolerance test with measurement of plasma insulin and amylin concentrations every 30 min for 2 h. Body composition and BMD/height (to provide a quantity with the dimensions of volumetric density that is independent of body size) were measured by dual-energy X-ray absorptiometry, and volumetric density of the third lumbar vertebral body was calculated. Circulating insulin concentrations correlated with BMD/height and volumetric density of the third lumbar vertebral body (r = 0.28-0.52). They also were related to body weight (r = 0.34-0.56) and fat mass (r = 0.38-0.56) but were not independently related to lean mass on multiple regression. There were no consistent relationships between amylin levels and these variables. Multiple-regression analyses with fat mass and insulin levels as independent variables indicated that BMD/height of total body and femoral trochanter were primarily related to fat mass, whereas, in femoral neck, the significant relationship was with insulin. Volumetric density of the third lumbar vertebral body was related to insulin levels alone on this analysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238342 TI - Pentoxifylline decreases body weight loss and muscle protein wasting characteristics of sepsis. AB - Sepsis induces metabolic disorders that include loss of body weight, muscle wasting, and acute-phase protein synthesis in liver. Cytokines are generally recognized as active mediators of these disorders, and the implication of tumor necrosis factor (TNF) has been frequently discussed in the recent past. However, the identity of the active agent in alterations of protein metabolism is still controversial. To improve our understanding of the role of cytokines in mediating muscle wasting observed in sepsis, we investigated muscle and liver protein metabolism in the following three groups of rats: infected control rats (INF-C); infected rats pretreated with pentoxifylline (PTX-INF), which is a potent inhibitor of TNF secretion; and pair-fed rats for the PTX-INF group pretreated with pentoxifylline. Pentoxifylline nearly completely suppressed TNF secretion but did not influence the transient fall in rectal temperature, the decreased hematocrit, and the increased liver protein mass and synthesis observed in INF-C rats. Pentoxifylline decreased the anorexia, the loss of body weight and muscle protein observed in INF-C animals, and partially prevented the decrease in muscle protein synthesis induced by infection. The overall data indicate that pentoxifylline is an effective agent in mitigating the characteristic muscle protein wasting induced by sepsis and confirm the limited role of TNF in the mediation of the acute-phase protein synthesis. Our results suggest a probable implication of TNF in the regulation of protein balance in muscle but do not allow discarding possible implication of other mediators that would be inhibited by pentoxifylline. PMID- 8238343 TI - Content, in vivo release, and bioactivity of fetal pulmonary immunoreactive adrenocorticotropin. AB - We hypothesized that fetal lung contains and releases immunoreactive (ir) adrenocorticotropin (ACTH). Fetal sheep lung (89-145 days gestation, n = 13) irACTH content was 4,100 +/- 672 (SE) pg/g lung wet wt compared with 5,425 +/- 1,403 (SE) pg/g in adults (n = 5) when measured by radioimmunoassay (RIA). irACTH values normalized to protein content were 158 +/- 18 and 95 +/- 22 (SE) ng/g protein for fetuses and adults, respectively. Grouped by age, < 90 days (n = 4), 131-139 days (n = 5), and 143-145 days (n = 4), the values for irACTH were 184 +/ 28, 173 +/- 33, and 114 +/- 33 (SE) ng/g protein, respectively, and declined significantly from < 90 days to adulthood. RIA serial dilution curve slopes were different from standard in 7 of 13 fetuses and in all adults, suggesting that irACTH structure differs from ACTH-(1-39). Acute in vivo fetal pulmonary artery and vein sampling (n = 5, 120-128 days gestation) revealed a mean difference of 609 +/- 476 (SE) pg/ml, a statistically significant release of irACTH across the lung. Western blotting revealed that pulmonary irACTH migrates in multiple immunostaining bands between molecular weight 14,000 and 46,000 and does not colocalize with ACTH-(1-39). Adrenal cell bioassay revealed that pulmonary irACTH is not corticotropic. We concluded that ovine fetal lung contains large-molecular weight irACTH, that content declines from 90 days gestation to adulthood, and that, under acute conditions, the fetal lung releases irACTH substances in sufficient quantity to contribute to circulating hormone concentrations of irACTH. PMID- 8238344 TI - Immunophysiology of the gut: a research frontier for integrative studies of the common mucosal immune system. AB - This review highlights work that, within the past decade, transformed mucosal immunophysiology from a hypothetical concept to a fully recognized interdiscipline. The regulation of epithelial and smooth muscle functions by the mucosal immune system represents an exquisitely sensitive adaptation to local antigenic challenge. Furthermore, immunologic cells communicate with nerves via paracrine secretions to rapidly transduce antigenic signals into panmucosal changes in function. These local immunocyte-nerve interactions are modulated by the autonomic and central nervous systems. Because of the common mucosal immune system, antigen-induced changes similar to those occurring in the intestine and colon are predicted to occur in mucosa of all hollow organs. The drawing together of fields as diverse as medicine and agriculture underscores the scope of areas encompassed by immunophysiology. Newly acquired knowledge has positioned the field to advance rapidly in both basic and applied directions. Forces that will remodel the field in the next decade will be derived from public concerns about human health maintenance and the explosive and novel use of new research tools stemming from molecular biology. These forces will draw on and advance our knowledge in areas as diverse as vaccine development and prevention of allergic reactions to foods, bioengineered foods in particular. PMID- 8238345 TI - PAP, a pancreatic secretory protein induced during acute pancreatitis, is expressed in rat intestine. AB - The pancreatitis-associated protein (PAP) is a lectin-related secretory protein present in small amounts in the rat pancreas and rapidly overexpressed during the acute phase of pancreatitis. We demonstrate in this report that PAP is also expressed in rat intestine. A cDNA library from rat jejunum was probed with pancreatic PAP cDNA. The inserts of the selected recombinant clones corresponded to a transcript whose nucleotide sequence was identical to that of pancreatic PAP mRNA. The transcript was detected in duodenum, jejunum, ileum, and colon. A protein with same molecular mass (16 kDa) and pI (8.2) as pancreatic PAP was actually immunodetected in ileum homogenate after separation by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Intestinal PAP was immunolocalized to the epithelial cells of the lower part of the villi. The protein accounted respectively for 0.02, 0.05, and 0.1% of soluble proteins in duodenum, jejunum, and ileum homogenates, as measured by enzyme-linked immunosorbent assay, and could not be detected in stomach and colon. Influence of fasting and feeding on PAP mRNA concentration was analyzed in ileum. Concentration decreased by 81 and 94% after animals were fasted for 24 and 48 h, respectively. Feeding restored the initial content within 6 h. On the other hand, intestinal PAP mRNA concentration was not altered during acute pancreatitis. PMID- 8238346 TI - Migrating spike complex in the small intestine of the fasting cat. AB - This study characterizes the migrating spike complex (MSC) in the small intestine of the awake fasting cat and compares the MSC with interdigestive activity in the small intestine of other species. Electrical activity in each of 12 cats with implanted electrodes showed MSCs, bands of spike potentials which attenuated slow wave frequency and amplitude as the MSCs progressed distally. MSCs occurred at variable frequency with intervals ranging from < 1 min to > 5 h and averaged 51.2 +/- 2.8 (SE) min. MSCs migrated at 1-8 mm/s, accelerating distally; the duration decreased distally such that the length of the bowel in a burst (2-3 cm proximally) was conserved. The MSC was associated with an intense prolonged contraction of duration similar to that of the MSC. Sometimes the MSCs occurred in close association, and when an MSC period was < 5.7 min, the second MSC propagated at a slower rate than the first. Frequently, a brief series of slow wave-associated spikes preceded an MSC. MSCs were not associated with slow waves. The MSC differs in several respects from the migrating myoelectric complex of other laboratory animals and is more appropriately classified in a category that includes giant migrating spikes, prolonged propagated contractions, power contractions, and migrating action potential complexes. PMID- 8238347 TI - Interstitial cells in deep muscular plexus of canine small intestine may be specialized smooth muscle cells. AB - The fine structures and properties of cells between the inner and outer circular muscle layers in the canine small intestine were studied by transmission electron microscopy (TEM), immunocytochemistry, and scanning electron microscopy (SEM). A nerve plexus (deep muscular plexus) supported by enteroglial cells, fibroblasts around blood vessels, macrophages, and thin and branched cells previously identified as interstitial cells of Cajal was observed. The interstitial cells of the deep muscular plexus (IC-DMP) were rich in mitochondria, dense bodies, and caveolae, and they were closely associated with nerve fibers. The IC-DMP had incomplete basal laminae. These cells also had numerous interconnecting gap junctions, and they also formed gap junctions with the surrounding smooth muscle cells of the outer circular muscle layer. IC-DMP were rich in myofilaments, which were primarily actin thin filaments, but myosin thick filaments, identified with anti-myosin light-chain antibodies, were also apparent. IC-DMP and circular smooth muscle cells both expressed immunoreactivity to anti-smooth muscle actin antisera, but these two types of cells differed in their intermediate filament proteins: IC-DMP featured vimentin immunopositive filaments, and circular smooth muscle cells featured desmin immunoreactivity. SEM showed that IC-DMP had thin and flat cell bodies with numerous branching processes. These cells came into close contact with nerve fibers and circular smooth muscle cells. The findings that IC-DMP cells contained myosin thick filaments and were immunopositive for anti-smooth muscle actin suggest that they may be more properly categorized as a type of smooth muscle cell. PMID- 8238348 TI - Gastric myoelectric and motor activity in dogs with gastric dilatation-volvulus. AB - Electrical and contractile properties of the stomach were assessed in six adult dogs after recovery from surgical treatment for gastric dilatation-volvulus (GDV), a disorder characterized by delayed gastric emptying of the solid phase. Electrodes and strain-gauge force transducers were sutured to the serosa of the antrum and pylorus at the time of surgical intervention for GDV. Ten days after implantation, electrical and mechanical activities were recorded before and after a standardized meal. The analog FM tape recordings of the electrical and mechanical signals were converted to digital time series for analysis by computer. Recordings from dogs after GDV showed increased slow wave propagation velocity in both the fasting and the fed states compared with controls. In addition, the GDV dogs had atypical fasting state phase III activity fronts. We found no difference in gastric slow wave frequency, dysrhythmia, or electromechanical coupling between the two groups. These results indicate that delayed gastric emptying in this syndrome is associated with increased gastric slow wave propagation velocity. PMID- 8238349 TI - Gastrointestinal transit is not impaired by regional loss of myenteric neurons in rat jejunum. AB - Chronic absence of myenteric neurons from a 5-cm segment of rat jejunum causes alterations in myoelectric activity. Spike potentials characteristic of phase III activity of the migrating motor complex (MMC) are present; however, the number of propagating spike potentials through the myenterically denervated region is reduced. The objective of this study was to examine the effect of the regional loss of myenteric neurons on gastrointestinal transit of a solid marker in the fasted rat. The rate of gastric emptying was not affected by the absence of the myenteric plexus in a 5-cm segment of the jejunum. However, 15 days after either myenteric denervation or vehicle treatment of a segment of jejunum, a more cephalad distribution and decreased rate of intestinal propulsion of the solid marker was observed in the small intestine. This delay in small intestinal transit observed at 15 days was not seen at 48 and 120 days. The decrease in transit at 15 days can be attributed to the handling of the bowel during the surgical procedure. The mouth-to-cecum transit time (MCT) was also not affected by chronic absence of the myenteric plexus. Furthermore, the MCT indicated that bacterial overgrowth, a common manifestation when gut motility is disrupted, did not occur in the small intestine after the experimental destruction of the myenteric plexus. The results of this study indicate that the regional loss of the myenteric plexus does not impair gastrointestinal transit in the fasted rat.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238350 TI - Nutrient feedback inhibition of gastric emptying plays a larger role than osmotically dependent duodenal resistance. AB - The slowing of gastric emptying by hyperosmolar solutions has been postulated to result from the triggering of duodenal osmoreceptor feedback on the stomach. We tested the idea that the inhibition of gastric emptying by a hyperosmolar solution depended on the duodenal resistance and the triggering of nutrient specific feedback by tracking gastric emptying of 300 and 1,200 mosmol/kgH2O test solutions in 12 dogs in which duodenal resistance was either removed (by temporarily diverting chyme from uncorked duodenal fistula) or preserved (by keeping duodenal fistula corked). Mannitol was used to test osmolality alone, and glucose was used to examine the combined effects of osmolality and nutrient specific inhibitory feedback. We found that: 1) the slowing effect of hyperosmolality was more marked with the duodenal resistance preserved (P < 0.05; analysis of variance), 2) the slowing effect of glucose was greater than that of mannitol for all conditions (P = 0.01; analysis of variance), and 3) the inhibitory effect of mannitol was localized to the duodenum. We conclude that inhibition of gastric emptying by hyperosmolar mannitol depended primarily on duodenal resistance, while the inhibitory effect of hyperosmolar glucose depended on nutrient-specific feedback on the stomach more than duodenal resistance. PMID- 8238351 TI - Proximal duodenal enterocyte transport: evidence for Na(+)-H+ and Cl(-)-HCO3- exchange and NaHCO3 cotransport. AB - The duodenum, in contrast to the jejunum, actively secretes HCO3- at a high rate, a process that protects the mucosa from acid/peptic injury. Our purpose was to define the mechanisms involved in HCO3- transport by studying the acid-base transport processes in isolated duodenal enterocytes. Individual rat duodenocytes, isolated by a combination of Ca2+ chelation and collagenase, attached to a collagen matrix were loaded with the pH-sensitive fluoroprobe 2',7' bis(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM), and intracellular pH was monitored by microfluorospectrophotometry. To identify Na(+) H+ transport, cells in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid 1) were pulsed with NH4Cl (40 mM) in the absence and presence of amiloride and 2) were removed of Na+. To examine Cl(-)-HCO3- exchange, Cl- was removed from Ringer HCO3- superfusate in the presence and absence of dihydro-4,4' diisothiocyanostilbene-2,2'-disulfonic acid (H2DIDS). The NaHCO3 cotransporter was studied by addition and subtraction of Na+ to amiloride-treated and Cl(-) depleted enterocytes perfused with Na(+)- and Cl(-)-free Ringer-HCO3- buffer with and without H2DIDS. Mammalian duodenocytes contain at least three acid-base transporters: an amiloride-sensitive Na(+)-H+ exchanger that extrudes acid, a DIDS-sensitive Cl(-)-HCO3- exchanger that extrudes base, and a NaHCO3 cotransporter, also DIDS sensitive, that functions as a base loader. These acid base transporters likely play a key role in duodenal mucosal HCO3- secretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238352 TI - Distribution of adenylate cyclase and GTP-binding proteins in hepatic plasma membranes. AB - Hepatic membrane subfractions prepared from control rats demonstrated forskolin (FSK)-stimulated adenylate cyclase activity in the basolateral (sinusoidal) but not apical (canalicular) plasma membrane. After bile duct ligation (BDL) for 12 or 24 h, there was an increase in FSK-stimulated adenylate cyclase activity in the apical membrane (54.2 +/- 3.9 pmol.mg-1 x min-1). The mechanism for this increase was explored further. ATP hydrolysis was found to be much higher in the apical than the basolateral membrane. Increasing the ATP levels in the assay enhanced apical membrane adenylate cyclase activity (10.5 +/- 0.2 pmol.mg-l.min 1); however, total adenosinetriphosphatase (ATPase) activity was not altered after BDL. Extraction of the apical membrane with bile acids or other detergents resulted in a two- to threefold increase in adenylate cyclase activity (30.6 +/- 3.6 pmol.mg-1 x min-1; detergent C12E8) This suggested that bile duct ligation was acting via the detergent-like action of bile acids to uncover latent adenylate cyclase activity on apical membranes. Further studies demonstrated that both BDL and detergent extraction also enhanced toxin-directed ADP-ribosylation of Gs alpha (cholera toxin) and Gi alpha (pertussis toxin) in the apical but not the basolateral membrane. After BDL, Gi alpha was found to be twofold greater in the apical membrane than the basolateral membrane. Immunoblotting using specific G protein antibodies further confirmed that apical membranes from control rats had a higher concentration of Gi1, 2 alpha and beta and slightly elevated levels of Gi3 alpha and Gs alpha compared with the basolateral membrane. The results demonstrate that adenylate cyclase and heterotrimeric GTP-binding proteins are present on the apical membrane, but measurement of their functional activity requires detergent permeabilization of apical membrane vesicles and is limited by the presence of high ATPase activity. PMID- 8238353 TI - Quantitative relationship between liquid bolus flow and laryngeal closure during deglutition. AB - We employed digital image analysis to assess the relative movements of the hyoid, larynx, and bolus as a function of liquid bolus volume (2-15 ml), and synchronized these measurements with intrapharyngeal manometry. Comparisons were performed of bolus head and tail movement in relation to the timing of hyoid movement, contact between the arytenoid and epiglottic cartilages, and intrabolus pressures. Bolus head movement in the distal pharynx, as determined from flow plots, was relatively rapid. Initial bolus tail movement in the distal pharynx was delayed proportional to bolus volume, and once initiated, was slower than bolus head movement. Laryngeal exposure time, defined as the interval between bolus head arrival and tail departure at the hypopharyngeal recording sensor, increased as a function of bolus volume. The time interval between the initial anterior hyoid movement and bolus arrival at the larynx, determined by concurrent plotting of hyoid excursion and bolus head position, was constant despite increasing bolus volume. Similarly, the interval between epiglottic-arytenoid contact and bolus arrival was constant despite increased bolus volume. The early phase of intrabolus pressure was temporally associated with posterior movement of the tongue base and varied as a function of bolus volume, whereas late intrabolus pressure was temporally associated with initial pharyngeal wall movement and was not significantly volume dependent. These data indicate that the temporal relationship between laryngeal closure and bolus head flow remains constant despite changes of laryngeal exposure time to the bolus as a function of volume. PMID- 8238354 TI - Modulation of uptake of heme by rat small intestinal mucosa in iron deficiency. AB - Although body iron status modulates whole body retention of heme derived iron, it is not known with certainty whether modulation occurs by regulation of mucosal uptake of heme. In vivo uptake from perfused intestine of heme labeled with 14C in the porphyrin ring was studied in groups of rats of differing iron status ranging from fully replete to markedly iron deficient. Heme extraction from infusate and mucosal heme uptake were significantly different between test groups (P < 0.005 and 0.001, respectively). Marked iron deficiency induced a 4.8-fold rise in heme extraction relative to iron-replete animals; rats with latent iron deficiency showed a smaller but still significant rise. Heme extraction correlated negatively with indicators of iron status: hemoglobin (r = -0.76, P < 0.001) and serum iron (r = -0.56, P < 0.05). The specific binding of [14C]heme to purified brush borders from iron-replete and iron-deficient rats was 1.4-fold higher in deficient rats when expressed per milligram of protein (P = 0.046) and 3.3-fold higher when expressed relative to alkaline phosphatase activity (P = 0.014). Thus mucosal uptake of heme in iron deficiency is increased because of an increase in its binding to the brush border. PMID- 8238355 TI - Secretin causes H+ secretion from intrahepatic bile ductules by vacuolar-type H(+)-ATPase. AB - Intrahepatic bile duct epithelial cells contribute to bile formation by hormone dependently secreting HCO3- to bile and H+ to periductular fluid. The present study was undertaken to determine whether the secretin-induced H+ secretion is due to activation of a H(+)-ATPase or Na(+)-H+ exchange. H+ secretion was estimated from the rate of intracellular pH (pHi) recovery after acid loading (24 mM NH4Cl) of microdissected bile ductules from pig liver mounted in a flow through chamber on the stage of a microscope. pHi was measured from an estimated average of 10-15 epithelial cells using the fluorescent pHi indicator 2',7' bis(carboxyethyl)-5,6-carboxyfluorescein and dual-wavelength excitation of fluorescence. The ducts were superfused with HCO3(-)-free N-2 hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffers. We found that secretin induced net H+ secretion of 4.53 +/- 0.7 mumol.ml cell volume-1 x min-1. This H+ secretion was blocked by 10(-6) M bafilomycin A1 but was unaffected by Na+ substitution with choline in the superfusion buffer. The experiments also showed that bafilomycin A1 did not block Na(+)-H+ exchange. The secretin-induced H+ secretion is probably caused by a vacuolar-type H(+)-ATPase and may constitute an important element of the cellular mechanisms causing secretin-dependent ductular HCO3- secretion into bile. PMID- 8238356 TI - Involvement of phospholipase D in caerulein-induced phosphatidylcholine hydrolysis in rat pancreatic acini. AB - Phosphatidylcholine (PC) metabolism stimulated by caerulein (Cae), a cholecystokinin analogue, was investigated in rat pancreatic acini prelabeled with [3H]choline or [3H]-myristic acid. Both labels were incorporated mostly into PC. An inhibition of choline incorporation into PC was first observed in response to Cae (100 and 500 pM) stimulation, as indicated by reduced [3H]choline incorporation into trichloroacetic acid-precipitable material. Whereas choline incorporation was reduced in PC, Cae (500 pM) significantly increased [3H]choline metabolites release in the incubation medium. Separation of these metabolites by thin-layer chromatography showed that approximately 90% of the labeled products released into the medium was phosphocholine; however, Cae caused significant increases of [3H]choline release after 5, 15, and 30 min. In response to Cae, manoalide, a phospholipase C (PLC) inhibitor, totally prevented phosphocholine release into the medium but did not affect choline release. Staurosporine, a protein kinase C inhibitor, did not influence basal and Cae-induced choline release. In cells prelabeled with [3H]myristic acid, Cae stimulated within 5 min a rapid increase in intracellular [3H]phosphatidic acid (PA) levels in the presence of the PA phosphohydrolase inhibitor, propranolol; this PA production was further increased after 15 and 30 min of stimulation. The time course of [3H]PA formation in the presence of propranolol was similar to that of choline release in the medium. Staurosporine partially blocked PA accumulation stimulated by Cae after 30 min. In contrast, manoalide significantly reduced basal PA accumulation but did not prevent its production in response to Cae. In the presence of ethanol, Cae also significantly stimulated above control values the formation of [3H]phosphatidylethanol. These data indicate that Cae-induced PC hydrolysis in rat pancreatic acini is mediated mostly by phospholipase D (PLD) to produce PA and choline; they suggest a direct action of Cae on PLD activation, an effect independent of PLC activation. PMID- 8238357 TI - Characterization of cytochrome P-450-dependent arachidonic acid metabolism in rabbit intestine. AB - We characterized cytochrome P-450-dependent arachidonate (P-450-AA) metabolism throughout the intestinal tract, since some metabolites derived via this pathway modify epithelial ion transport and regional blood flow. Microsomes (0.3 mg/ml) were prepared from each region of the intestines of anesthetized New Zealand White male rabbits and incubated with [14C]AA (7 microM) for 30 min at 37 degrees C. In the presence of NADPH (1 mM), ileal microsomes exhibited the greatest P-450 AA metabolism, whereas duodenal microsomes exhibited little or no activity. For jejunal, ileal, and cecal microsomes, AA metabolism was reduced in the absence of NADPH and by boiling microsomes, was unaffected by indomethacin (10 microM) and BW-755C (50 microM), but was significantly attenuated by the P-450 enzyme inhibitors, 7-ethoxyresorufin (1 microM) and SKF-525A (100 microM). However, colonic (ascending, transverse, and descending) microsomal activity was inhibited by both P-450 and lipoxygenase inhibitors. Analysis of ileal AA metabolites by high-pressure liquid chromatography and negative ion chemical ionization gas chromatography-mass spectrometry revealed products corresponding to monohydroxyeicosatetraenoic acids (HETEs). Semiquantitative analysis showed that 20-, 19-, 18-, 17-, and 16-HETEs were present in a ratio of 6.2:3.3:0.3:0.1:0.1, respectively. Furthermore, ileal P-450-HETEs dilated the isolated perfused mesenteric bed, as did 20-HETE, the predominant ileal AA metabolite. Because 20 HETE was also shown to affect epithelial ion transport, we suggest that P-450-AA metabolites may make important contributions to intestinal function. PMID- 8238358 TI - Effects of the somatostatin analogue octreotide on rectal afferent nerves in humans. AB - Somatostatin (Som) administered intrathecally to humans has been shown to exert potent analgesic effects on somatic pain, and anecdotal evidence suggests that Som may also relieve visceral pain. In the current study, we used rectal balloon distension in seven healthy volunteers to evaluate the effect of the Som analogue octreotide (Oct; 1.25 microgram/kg sc) on four pathways mediated by different visceral afferents that originate in the rectum: conscious perception, receptive relaxation, reflex inhibition of internal anal sphincter, and a viscerosomatic reflex. Rectal mechanoreceptors were stimulated either by distending the rectum tonically (volume ramp at 20-40 and 400 ml/min) or phasically (intermittent pressure steps of 60 s duration). Pressure thresholds for nonnoxious and noxious sensations in response to slow tonic distension were increased in the presence of rectal lidocaine (20 ml of 2% solution), whereas those to phasic distension were unaffected. Oct significantly increased pressure and volume thresholds for nonnoxious and noxious sensations in response to slow tonic distension but did not further increase thresholds in the presence of intrarectal lidocaine. In contrast, no effect of Oct on rectal sensations was observed during rapid tonic or phasic distension. Oct had no effect on any of the monitored reflex responses. The effect of Oct on rectal sensation in the concentration used in this study was not associated with changes in the rectal wall pressure-volume relationship during any distension protocol. These findings indicate that the inhibitory effect of Oct on rectal sensation is likely to represent a direct effect on a subset of extrinsic primary afferent neurons, with receptive fields in the mucosa. PMID- 8238359 TI - Role of histamine2 receptor in increased expression of rat gastric H(+)-K(+) ATPase alpha-subunit induced by omeprazole. AB - Omeprazole is a specific inhibitor in vivo of the functioning gastric acid pump, the H(+)-K(+)-adenosinetriphosphatase (ATPase), in the secretory canaliculus of the parietal cell. It has been shown previously that omeprazole in rats led to an increase in the mRNA for the alpha-subunit of the H(+)-K(+)-ATPase. Omeprazole causes a marked increase in circulating gastrin in this species, which in turn stimulates release of histamine from the enterochromaffin-like cell. The possible role of this pathway was investigated by the in vivo administration of famotidine, a potent H2 receptor antagonist. A single intraperitoneal dose of famotidine, 200 mg/kg, produced a transient hypergastrinemia peaking at 3 h and normalizing at 12 h, inhibition of secretion that lasted for 12 h, but no change in the level of the alpha-subunit mRNA or of beta-actin mRNA. In contrast, a single dose of omeprazole, 100 mg/kg, inhibited acid secretion and produced hypergastrinemia, peaking at 12 h, both effects lasting for the 24-h observation period. Omeprazole elevated the alpha-subunit mRNA transiently by more than threefold at 3 h, with normal levels being restored at 24 h. The administration of famotidine 1 h after omeprazole did not change the effects of omeprazole on acid secretion but elevated the gastrin levels further. There was now no elevation of the alpha-subunit mRNA for the first 6 h, but a small increase at 12 h and a further increase to approximately 2.5-fold at 24 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238360 TI - Effects of gastric pacing on canine gastric motility and emptying. AB - Gastric pacing has been achieved in dogs and humans, but its effects on gastric motility and emptying have not been thoroughly explored. Seven dogs had bipolar electrodes placed 1 and 10 cm proximal to the pylorus for reverse and forward pacing and monopolar recording electrodes and strain gauges placed 3, 5, and 7 cm proximal to the pylorus. After recovery, myoelectrical and contractile activity and gastric emptying of a mixed meal (50 g 99mTc-labeled liver and 250 ml 111In labeled 5% dextrose broth solution) were measured in each of three conditions: no pacing, reverse pacing, and forward pacing (frequency 0.5 cycles/min above intrinsic pacesetter potential frequency). Reverse pacing reversed the direction of > 90% of antral pacesetter potentials and peristaltic waves in six of seven dogs, prolonged the lag phase of solid emptying, prolonged the half emptying time of solids and liquids, and increased the antral motility index. Forward pacing entrained pacesetter potentials but had no consistent effect on emptying or antral contractions. In conclusion, reverse gastric pacing slows gastric emptying of digestible solids and liquids by reversing the direction of antral peristalsis and increasing the antral motility index, whereas forward pacing has no such effects. PMID- 8238361 TI - Na(+)-K(+)-ATPase gene expression in rat intestine and Caco-2 cells: response to thyroid hormone. AB - Expression of the Na(+)-K(+)-adenosinetriphosphatase (ATPase) gene family in rat intestinal epithelial cells was examined using RNA blot hybridization analyses. Rat intestinal epithelial cells express only the alpha 1- and beta 1-subunit mRNAs. A gradient in expression of alpha 1- and beta 1-subunit mRNA was seen along the villus-crypt unit in both jejunum and ileum, i.e., villus tip >> crypt cells. Regional differences in expression were observed along the intestine. alpha 1- and beta 1-subunit mRNA abundance was similar in jejunum, ileum, and colon while enzymatic activity was highest in the jejunum and lowest in the ileum. Administration of thyroid hormone to thyroidectomized rats increased the expression of alpha 1- and beta 1-subunit mRNAs in jejunum but not in colon. Hypothyroidism had no effect on subunit mRNA expression. The human intestinal cell line Caco-2 was also studied. These cells also expressed only the alpha 1- and beta 1-isoform mRNAs and demonstrated a developmental profile in both mRNA and enzymatic activity. Furthermore, in Caco-2 cells both alpha 1- and beta 1 mRNAs and Na(+)-K(+)-ATPase enzymatic activity were stimulated by thyroid hormone. Caco-2 cells transfected with 5' flanking regions of the human Na(+) K(+)-ATPase beta 1-gene linked to the chloramphenicol acetyltransferase (CAT) reporter gene responded to 3,5,3'-triiodothyronine (T3) treatment with increased expression of CAT activity. This suggests that the 5' flanking region of the beta 1-gene contains a thyroid hormone response element and that T3 upregulation occurs at the transcriptional level.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238362 TI - Effect of ursodeoxycholic acid on intracellular pH regulation in isolated rat bile duct epithelial cells. AB - To determine if ursodeoxycholic acid (UDCA) induces a HCO3(-)-rich hypercholeresis by stimulating HCO3- secretion from bile duct epithelial (BDE) cells, we studied the effect of UDCA, sodium tauroursodeoxycholate (TUDCA), and cholic acid on intracellular pH (pHi) regulation and HCO3- excretion in BDE cells isolated from normal rat liver. Exposure of BDE cells to UDCA (0.5-1.5 mM) produced a dose-dependent initial acidification [from -0.05 to -0.16 pH units (pHu)], which was lower in Krebs-Ringer bicarbonate than in N-2 hydroxyethylpiperazine-N'-2- ethanesulfonic acid (HEPES), because of the higher cell-buffering power in the presence of HCO3-. In contrast, TUDCA (1 mM) had no effect on pHi in either media. BDE acidification induced by UDCA (1.5 mM) in KRB was not inhibited by Cl- depletion excluding activation of Cl(-)-HCO3- exchange. Most BDE cells spontaneously recovered their basal pHi during the UDCA infusion (0.5-1 mM) by a secondary activation of the Na(+)-H+ exchanger (amiloride inhibition of pHi recovery; n = 4), and pHi overshot basal levels by 0.1-0.2 pHu after UDCA withdrawal. The activity of Cl(-)-HCO3- exchange (Cl- removal/readmission maneuver) as well as the activities of Na(+)-H+ exchange and Na(+)-HCO3- symport (NH4Cl acid load in HEPES and KRB, respectively) were unaffected by UDCA (0.5 mM) compared with controls. Cholic acid (1.5 mM), which does not produce a hypercholeresis, also acidified BDE cells in KRB media. These studies indicate that UDCA does not stimulate HCO3- excretion from isolated rat BDE cells but modifies pHi in BDE cells as a weak acid. PMID- 8238363 TI - Mechanical properties of isolated smooth muscle from human rectum and internal anal sphincter. AB - The passive and active length-tension relations of the circular smooth muscle layer of the human distal rectum and the proximal and distal internal anal sphincter were investigated. Muscle strips were prepared and mounted in organ baths for recording of isometric tension. Resting lengths (LR) were measured, and the preparations were elongated stepwise. At each length, the corresponding values for passive tension, spontaneous active resting tension, and the submaximal active tension were recorded. Elongations of 200-380% of LR were possible before a sharp increase in passive tension occurred. None of the mean tension values measured at length for maximal active tension (LO) differed significantly among the three muscle types. All strips developed active resting tension. This tension was myogenic and contributed 10 +/- 3, 23 +/- 6, and 27 +/- 6% to the total active performance of rectal and proximal and distal sphincter preparations, respectively. Collagen constituted approximately 50% of smooth muscle biopsies, with highest contents in distal internal anal sphincter. This study provides an acceptable method for assessing the optimal experimental length by stretching the strips in an inactive state to 200% of LR, followed by individual adjustment of the passive tension to 5 mN/mm2 measured at 200% of LR. PMID- 8238364 TI - Inhibitory effect of CO on internal anal sphincter: heme oxygenase inhibitor inhibits NANC relaxation. AB - We examined the effect and role of CO in opossum internal anal sphincter (IAS) relaxation in response to nonadrenergic noncholinergic (NANC) nerve stimulation. Effects of NANC nerve stimulation on the IAS tension and second messengers (cAMP and cGMP) were examined before and after the selective heme oxygenase (HO) inhibitor zinc protoporphyrin IX (Zn PP-IX). The HO activity of the IAS smooth muscle was determined before and after NANC nerve stimulation. CO caused a concentration-dependent and tetrodotoxin-resistant fall in the resting tension of the IAS. The direct action of CO was confirmed by its relaxant action on the isolated smooth muscle cells. Furthermore, CO caused an increase in the tissue cGMP levels comparable to that observed with nerve stimulation. Zn PP-IX caused suppression of IAS relaxation caused by NANC nerve stimulation and vasoactive intestinal polypeptide (VIP) but not by peptide histidine-isoleucine and suppression of the increase in cGMP in response to NANC nerve stimulation. Zn PP IX had no significant effect on the IAS responses to CO, nitric oxide (NO), and the beta-adrenoceptor agonist isoproterenol. The IAS responses to CO were not modified by the NO synthase inhibitor NG-nitro-L-arginine. Significant HO activity was detected in the IAS, which increased further in response to NANC nerve stimulation and VIP. The direct relaxant actions of CO and the suppression of NANC-mediated relaxation of the IAS by the HO inhibitor suggest the involvement of CO in the neurally mediated IAS relaxation. PMID- 8238365 TI - Postnatal lung growth: evidence that the gas-exchange region grows fastest at the periphery. AB - The site(s) at which the gas-exchange region enlarges after alveoli are formed from the saccules of the immature lung is unclear; however, this information might be important to studies of the regulation of lung growth. Although aware of important assumptions on which it rests, we undertook this study to test the idea that the lung's gas-exchange region enlarges more rapidly in the immediate subpleural region than more centrally. To label the interstitium of the gas exchange region, rats were provided silver-containing water from age 23 to 135 days (112 days). Some were killed at age 135 days, others 51 days after silver exposure ended (age 186 days). We considered silver grains that formed in the interstitium as a marker of lung present or formed during silver exposure; tissue added after exposure would diminish the numerical density of grains and the fastest growing sites would have the lowest grain density. Rats killed on the 112th day of silver exposure had a silver grain density in their immediate subpleural gas-exchange matrix that was 28% lower than in their more central gas exchange matrix. Rats killed 51 days after silver exposure ended had a grain density in the immediate subpleural region that was 65% lower than in the central matrix of the gas-exchange region. The grain density in the rats killed 51 days after silver exposure ended was 33% lower in the central matrix and 67% lower in the peripheral matrix than in the respective regions of rats killed on day 112 of exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238366 TI - G protein-regulated large-conductance chloride channels in freshly isolated fetal type II alveolar epithelial cells. AB - Using the patch-clamp technique, we have recorded single channels in cell attached and inside-out excised patches from the plasma membrane of type II alveolar epithelial cells freshly isolated from fetal guinea pig lung by elastase digestion and differential filtration. In cell-free patches the channels were highly selective for Cl- (PCl:Pcat = 9:1), had a large unitary conductance (375 pS +/- 23 pS), and current reversal of 0 mV in either symmetrical Na(+)-rich solutions or when the inner membrane leaflet was bathed in a K(+)-rich solution. The large-conductance Cl- channel exhibited little or no voltage inactivation at positive potentials, remained open for a significant amount of time at potentials negative to -40 mV, and was blocked at all potentials by 0.1 mM 4-acetamido-4' isothiocyanostilbene-2,2'-disulfonic acid. Channel activity was independent of intracellular calcium concentration. Bath addition of the nonmetabolizable analogue of GTP, GTP gamma S (0.1 mM), caused a voltage-dependent inhibition of channel activity [open probability (Po) plot was shifted by at least +25 mV]. Smaller channels (25 +/- 3 pS) were recorded in the cell-attached configuration with a current-voltage (I-V) relationship which was compatible with a Cl- conductance. On excision, the patches previously containing small-conductance channels exhibited only large-conductance Cl- channel behavior. These large conductance, G protein-regulatable Cl- channels may provide a route for alveolar cell Cl- exit and as such may be an integral part of the mechanism responsible for secretion of fetal lung fluid. PMID- 8238367 TI - Uptake of antioxidants in surfactant liposomes by cultured alveolar type II cells is enhanced by SP-A. AB - Antioxidant delivery may be targeted toward the alveolar epithelium by encapsulating superoxide dismutase (SOD) and catalase in liposomes made from pulmonary surfactant. We studied whether antioxidant-surfactant liposomes increase cellular antioxidant activity in alveolar type II cells and whether this effect is influenced by the presence of surfactant protein A (SP-A). Cu,Zn SOD and catalase were encapsulated in liposomes made from synthetic phospholipids with or without 5% SP-A or from natural cow surfactant. Alveolar type II cells from adult rats were preincubated for 20 h, and liposome mixtures were added for 24 h, followed by measurement of cellular SOD and catalase activities (U/mg DNA). Antioxidant-surfactant liposomes increased alveolar type II cell antioxidant activity sharply. Uptake of SOD/catalase from liposomes with synthetic phospholipids and SP-A was twice that from liposomes without SP-A and did not further improve in the presence of SP-B and -C. Encapsulation of antioxidants diminished the surface activity of the surfactant liposomes, but this feature was absent in the presence of SP-A. These data suggest that: 1) antioxidant surfactant liposomes augment alveolar type II cell antioxidant activity, 2) liposomal uptake is facilitated by the presence of SP-A, and 3) inhibition of surface activity of surfactant by encapsulated antioxidants can be reversed by SP A. PMID- 8238368 TI - Mitigation of oxidant injury to lung microvasculature by intratracheal instillation of antioxidant enzymes. AB - We quantitated the ability of intratracheally administered liposome-encapsulated antioxidant enzymes to reduce reactive oxygen species injury to the pulmonary microvasculature. Cationic liposomes containing 3,500 U of Cu,Zn superoxide dismutase (Cu,Zn SOD) and 3,124 U of catalase were instilled into rabbits. The animals were killed 2-72 h later and their lungs were removed and perfused with Krebs Ringer with 5% wt/vol of fat-free bovine serum albumin. The pulmonary filtration co-efficient (Kf,c) was measured before and after adding 500 microM xanthine and 5 mU/ml xanthine oxidase (XO) into the lung perfusate. Two hours after a single intratracheal instillation of liposome-entrapped Cu,Zn SOD and catalase, lung antioxidant enzyme activities were 34 and 125% higher than the corresponding control values, remained virtually unchanged for up to 8 h post instillation, and then decreased, reaching baseline values between 24 and 72 h. Addition of xanthine and XO into the lung perfusate of un-instilled rabbits, or rabbits that received liposomes with inactivated enzymes, caused a 100% increase in Kf,c (control value: 2 +/- 0.12 ml.min-1 x cmH2O-1 per 100 g dry lung weight). On the other hand, Kf,c values of rabbits lungs instilled with liposome encapsulated active Cu,Zn SOD and catalase and challenged with xanthine and XO 8 24 h later remained at baseline levels. Instillation of liposomes containing either enzyme was equally effective in preventing the increase in Kf,c, indicating that both superoxide anions and hydrogen peroxide were necessary for the initiation of injury. We concluded that intratracheal instillation of liposome-encapsulated antioxidant enzymes caused a transient increase of lung antioxidant enzyme levels which protects the pulmonary microvasculature from free radical-initiated injury. PMID- 8238369 TI - Receptor mechanism of thrombin-mediated pulmonary vasodilation in neonates. AB - A recently identified peptide sequence exposed after proteolytic cleavage of the NH2-terminus of the thrombin receptor mimics some cellular effects of alpha thrombin. To determine whether a proteolytic action of thrombin is required for vasoactivity, we examined the vascular effects of modified thrombins and synthetic NH2-terminus peptide sequences of the thrombin receptor (TRPs) in isolated piglet lungs. Lungs of piglets 1-6 days old were perfused with recirculating Ringer-albumin solution at a constant flow of 60 ml/min. We measured the pulmonary artery pressure (Ppa) and segmental distribution of pulmonary vascular resistance (using the double occlusion method) in response to injections of human alpha-thrombin, modified thrombins, and TRP-14 and TRP-7 (i.e., 14 and 7 amino acid NH2-terminus peptides of the cleaved thrombin receptor). alpha-Thrombin produced a rapid and transient decrease in Ppa; the magnitude and duration [time for one-half recovery (t1/2 R)] of the vasodilation responses were concentration dependent [t1/2 R values of 1.4 +/- 0.1 and 3.3 +/- 2.4 min (mean +/- SE) at concentrations of 10(-10) and 10(-9) M, respectively]. The vasodilation was due primarily to a decrease in precapillary resistance. Proteolytically active, but binding-impaired gamma-thrombin was a less potent vasodilator and proteolytically inactive D-phenylalanyl-prolyl-arginine chloromethyl ketone (PPACK)-alpha-thrombin did not induce vasodilation. TRP-14 was also a pulmonary vasodilator with a t1/2R value of 0.8 +/- 0.09 min at a concentration of 10(-7) M; both TRP-14 and TRP-7 were approximately 3-log less potent than equimolar alpha-thrombin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238370 TI - Paraquat-induced lung injury: prevention by vasoactive intestinal peptide and related peptide helodermin. AB - We earlier showed that the neuropeptide vasoactive intestinal peptide (VIP) reduces or prevents acute injury produced in rat lungs by xanthine and xanthine oxidase. We have now examined whether VIP can protect against lung injury induced by paraquat, a prooxidant pesticide. Isolated guinea pig lungs were perfused for 60 min with Krebs-4% albumin and mechanically ventilated with 95% O2-5% CO2. Infusion of paraquat (100 mg/kg) into the pulmonary artery (n = 9 observations) increased peak airway pressure from 10.1 +/- 0.6 to 54.7 +/- 6.5 cmH2O, perfusion pressure from 8.0 +/- 0.5 to 14.9 +/- 3.0 cmH2O, wet-to-dry lung weight ratio to 7.17 +/- 0.37, and bronchoalveolar lavage protein content to 2.70 +/- 0.83 mg/ml (P < 0.01). Pretreatment with 1-3 micrograms.kg-1 x min-1 VIP markedly attenuated or prevented all abnormalities. Of the related peptides tested, helodermin was as effective as VIP, but secretin and glucagon were ineffective. The results demonstrate that VIP and helodermin protect perfused guinea pig lungs against paraquat-induced injury and support the view that VIP has antioxidant activity. PMID- 8238371 TI - GLP-1 stimulates secretion of macromolecules from airways and relaxes pulmonary artery. AB - Recent data revealed the existence of specific receptors for glucagon-like peptide-1(7-36)amide (GLP-1) on rat lung membranes. Utilizing slide-mount autoradiography of fresh frozen lung tissue sections, we have localized binding sites for GLP-1 on mucous glands in the trachea and on vascular smooth muscle of the pulmonary artery. When tracheas were incubated in a modified Ussing chamber, the addition of GLP-1 to the submucosal side increased 35S-sulfate-labeled macromolecule secretion (191 +/- 12% above basal, P < 0.005). The optimal secretory response elicited by GLP-1 was approximately 23% of the maximal secretory response after a maximal acetylcholine stimulation. Other proglucagon derived peptides such as glucagon, oxyntomodulin, and GLP-2 had no effect. In isolated rings of arteries, GLP-1 (10(-8) to 10(-5) M) induced a dose-dependent and time-reversible relaxation of preconstricted arteries. In a preparation with denuded epithelium, GLP-1 lost its effect. In conclusion, GLP-1 might represent another neuropeptide that acts as neurotransmitter of the peptidergic, nonadrenergic-noncholinergic nervous system that innervates the airways. PMID- 8238372 TI - Pleural mesothelial cell response to inflammation: tumor necrosis factor-induced mitogenesis and collagen synthesis. AB - This study examined the effects of tumor necrosis factor-alpha on pleural mesothelial cell proliferation and collagen synthesis, functions which may be important in the response of the pleura to injury. Tumor necrosis factor-alpha caused a significant increase in proliferation and collagen production by rat pleural mesothelial cells in vitro. Proliferation increased in a time- and dose dependent manner, resulting in an approximate twofold increase in the uptake of [3H]thymidine relative to control. The uptake of [3H]proline into collagenase sensitive protein increased in a dose-dependent manner for concentrations of tumor necrosis factor-alpha > or = 1.0 ng/ml. The increase in collagen production were associated with increased steady-state levels of alpha 1(I)-procollagen mRNA. These results suggest that tumor necrosis factor-alpha may have a significant effect on pleural mesothelial cell function in vivo in the setting of inflammation. Increases in pleural mesothelial cell proliferation and collagen synthesis in response to inflammatory mediators, like tumor necrosis factor alpha, may be important in healing the pleura after injury by a variety of disease processes. PMID- 8238373 TI - Visualization of high- and low-affinity beta-adrenergic receptors in rat lung: upregulation by chronic hypoxia. AB - Chronic hypoxia induces a hyperadrenergic state which down-regulates beta adrenergic receptors (beta-AR) in the heart. We visualized lung beta-AR binding sites after adaptation to chronic hypoxia by autoradiography of whole lung sections labeled with 50 pM 125I-labeled pindolol. A low concentration of agonist (32 nM isoproterenol) selectively masked beta-ARs with high affinity for agonists. Total specific beta-AR binding increased twofold with hypoxia. In both the control and hypoxic lung sections, 60-70% of the beta-ARs were in a high affinity state, which could be reversed by guanine nucleotide. Autoradiography revealed a high density of high- and low-affinity beta-AR sites in lung parenchyma, predominantly involving alveolar walls, but also the walls of airways and blood vessels. The distribution of high- and low-affinity beta-AR within the lung was qualitatively identical. Hypoxia increased beta-AR binding without affecting its distribution. The majority of the additional beta-ARs induced during adaptation to chronic hypoxia are in the high-affinity state, and are thus of probable functional significance. PMID- 8238374 TI - Oxygen conformance of cellular respiration in hepatocytes. AB - Cellular respiratory rates are normally determined by metabolic activity, but become rate limited by O2 availability if the cell O2 tension (PO2) falls below a critical value (typically 1-10 Torr). An ability to reduce metabolic activity and energy demand in response to a falling O2 availability might confer an increased resistance to a diminished O2 supply. Isolated rat hepatocytes were studied in primary culture under controlled O2 tensions. Cells were obtained by collagenase digestion and seeded into nutritive media in control and experimental spinner flasks at identical cell densities. Cells subjected to rapid reduction in PO2 (100-->0 Torr over < 40 min) exhibited undiminished O2 uptake until PO2 fell below 10 Torr. By contrast, when cell PO2 was reduced over several hours, significant decreases in O2 uptake became evident at O2 tensions as high as 70 Torr. These decreases were associated with a reduction in ATP concentration and an increase in NAD(P)H, compared with rapidly deoxygenated cells at the same PO2. No loss in cell viability was detected after 24 h at reduced PO2. The decrease in respiratory rate was associated with an increased rate of lactic acid production relative to normoxic controls. Restoration of normoxia was associated with an immediate return of O2 uptake to control levels. These results demonstrate that hepatocytes are capable of reversibly decreasing metabolic activity and O2 demand during sustained moderate reductions in PO2, via a mechanism that appears to involve an inhibition of mitochondrial function other than O2 supply limitation. This response may alter cellular susceptibility to physiological stresses including hypoxia. PMID- 8238375 TI - Changes in lung expansion alter pulmonary DNA synthesis and IGF-II gene expression in fetal sheep. AB - Our aim was to determine the effect of short-term (7 days) alterations in fetal lung liquid volume on pulmonary DNA synthesis rates and insulin-like growth factor-II (IGF-II) mRNA levels. Fifteen chronically catheterized fetal sheep were divided into three groups. In one, the trachea was obstructed, in another lung liquid was drained by gravity, and the third group served as controls. After 7 days, [3H]thymidine was injected into each fetus and 8 h later fetal tissues were collected. Fetal lung-to-body weight ratios and total lung DNA contents were greatly increased in fetuses with tracheal obstruction compared with control fetuses, whereas the drainage of lung liquid did not affect these measurements. DNA synthesis rates in pulmonary tissue were significantly reduced from a mean control value of 153.3 +/- 25.1 disintegrations per minute (dpm)/microgram DNA to 57.2 +/- 8.6 dpm/microgram DNA by lung liquid drainage (P < 0.05) and were significantly increased to 236.0 +/- 24.0 dpm/microgram DNA by tracheal obstruction (P < 0.05). Following tracheal obstruction, lung IGF-II mRNA levels were increased to 177.0 +/- 18.2% (P < 0.05) of the mean value for control fetuses, whereas they were reduced to 56.1 +/- 7.1% of control in lung liquid drained fetuses. We conclude that altering fetal lung expansion has a potent and rapid effect on pulmonary DNA synthesis and that this effect may, in part, be mediated by an alteration in IGF-II gene expression. PMID- 8238376 TI - Modulation of vasoactive intestinal peptide pulmonary relaxation by NO in tracheally superfused guinea pig lungs. AB - The mechanism of vasoactive intestinal peptide (VIP)-induced pulmonary relaxation in tracheally perfused guinea pig lungs was defined with the use of inhibitors of nitric oxide synthase (NOS) and by direct measurement of nitric oxide (NO) equivalents recovered from lung perfusion fluid. Lungs treated with 200 microM NG nitro-L-arginine were resistant to the relaxant effects of VIP in these lungs; the 50% inhibitory dose (ID50) for VIP was 32 nmol/kg (95% confidence interval, 16-79), which was approximately 100-fold greater than the ID50 of control lungs which was 0.39 nmol/kg, (0.16-0.79, P < 0.0001). This inhibitory effect could be overcome with excess L- but not D-arginine. In contrast, VIP-induced relaxation of isolated guinea pig trachea was not modified by inhibitors of NOS. To confirm that VIP infusion resulted in NO generation in whole lungs, we measured NO equivalents in lung effluent by two distinct technologies. We found that VIP injection caused a significant increase in NO equivalents from 0.11 +/- 0.04 microM to 0.78 +/- 0.15 microM (P < 0.05) and that this increase preceded VIP induced pulmonary relaxation. Lungs pretreated with the putative guanylyl cyclase inhibitor methylene blue were less responsive to VIP [ID50 4.0 nmol/kg (1.5-10), P < 0.005 compared with control lungs], consistent with a physiologically significant guanosine 3',5'-cyclic monophosphate-dependent mechanism. Our data demonstrate that VIP has the capacity to relax whole lungs in part by stimulating the generation of NO. PMID- 8238377 TI - Renal renin-angiotensin system in diabetes: functional, immunohistochemical, and molecular biological correlations. AB - Recent evidence indicates a role for the renin-angiotensin system (RAS) in the pathogenesis of glomerular injury in diabetes. To further explore the RAS in diabetes, studies were conducted in nondiabetic control rats and in moderately hyperglycemic diabetic (DM) rats. In DM rats, both acute and chronic therapy with the specific angiotensin II (ANG II) receptor antagonist losartan did not affect glomerular hyperfiltration or hyperperfusion but selectively normalized the glomerular capillary hydraulic pressure and ultrafiltration coefficient. To determine the basis of intrarenal hemodynamic responsiveness to RAS inhibition, we conducted biochemical, molecular biological, and immunohistochemical studies to assess endogenous RAS activity. Values for plasma renin concentration and serum angiotensin-converting enzyme (ACE) activity in DM rats were normal. In contrast, intrarenal renin protein content, and renin and angiotensinogen mRNAs, were increased in DM rats, suggesting disproportionate activation of the intrarenal RAS. Total renal ACE activity was significantly reduced in DM rats, but immunohistochemical studies indicated redistribution of ACE in the diabetic kidney. Proximal tubule ACE activity was reduced, but ACE immunostaining intensity was enhanced in glomeruli and renal vasculature. Together, these observations indicate increased RAS activity in those sites (glomeruli and vasculature) most likely to regulate hemodynamic function, potentially explaining the prominent responses to pharmacological blockade of ANG II formation and/or action. PMID- 8238378 TI - Oxytocin affects apical sodium conductance in rabbit cortical collecting duct. AB - The physiological role of oxytocin (OT) in the kidney is still unclear, although autoradiographic data have shown the existence of OT receptors in the rat kidney. We examined the effect of OT in the microperfused rabbit cortical collecting duct (CCD) by using conventional cable analysis and microscope photometry. On addition of 10(-9) M OT to the bath, the lumen-negative transepithelial voltage (VT) transiently increased and the transepithelial resistance (RT) and the fractional resistance of the apical membrane (FRA) (1st phase) both decreased. After this initial change, the lumen-negative VT gradually decreased below its baseline level and RT and FRA (second phase) both increased. These electrical changes were dose dependent and were prevented by the addition of 10(-5) M amiloride to the lumen. Although responses to OT were not prevented by 10(-9) M arginine vasopressin (AVP) or 10(-6) M of a V1-receptor antagonist (OPC-21268) or V2 receptor antagonist (OPC-31260), they were inhibited by the addition of the specific OT antagonist des-Gly-NH2-[d(CH2)3,Tyr(Me),Thr]OVT. Additional studies of intracellular free calcium ([Ca2+]i) revealed that 10(-8)-10(-6) M OT caused an increase in [Ca2+]i in CCD in a dose-dependent manner. Also, pretreatment with 2 x 10(-8) M bis-(aminophenoxy)ethane-tetraacetic acid-acetoxymethyl ester, an intracellular Ca2+ chelator, abolished the electrical and [Ca2+]i responses to OT. Pretreatment with 5 x 10(-4) M 8-(4-chlorophenylthio)-adenosine 3',5'-cyclic monophosphate (CPT-cAMP) partially prevented the electrical responses to OT, thus reducing the decrease in lumen-negative VT below its basal level and the increase in RT after the 1st phase. These data show that OT affects the apical Na+ conductance of collecting duct cells through OT receptors distinct from the AVP receptors and that the effect of OT may, at least in part, be brought about by a mechanism(s) dependent on the increase in [Ca2+]i and cAMP production. PMID- 8238379 TI - A potential role for endogenous adenosine in control of human glomerular and tubular function. AB - Adenosine has profound effects on renal function in experimental animals, but little is known about its role in human subjects. The recent advent of specific adenosine agonists and antagonists suitable for human use, however, now makes it possible to evaluate the influence of this potent vasoactive compound in both normal and pathological states. In this study we assessed the effects of FK-453, a nonxanthine, selective adenosine A1-receptor antagonist, on normal renal hemodynamics, tubular function, and plasma renin release. Eight healthy, male subjects each received three single oral doses of FK-453 (50, 100, and 200 mg) in ascending dose order with random allocation of one matched placebo dose, each on a separate study day. Renal hemodynamics, tubular function, and plasma renin concentrations (PRC) were assessed at baseline and postdose on each study day. Glomerular filtration rate (clearance of 51Cr-labeled EDTA) rose by 18.0%, 3 h after the administration of 100 mg of FK-453 and by 18.3% and 23.5%, 2 and 3 h, respectively, after the 200-mg dose, which was significantly different from the changes following placebo. There were no significant changes in mean arterial blood pressure or effective renal plasma flow (clearance of 125I-Hippuran). In contrast there were statistically significant increases in urine flow rate and osmolar clearance, as well as absolute and fractional excretions of sodium, phosphate, bicarbonate, chloride, magnesium, and uric acid in response to FK-453. No glycosuria or aminoaciduria was detected on urinalysis. There was, in addition, a marked increase in PRC in response to FK-453.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238380 TI - Regulation of Na(+)-3HCO3- cotransport in rabbit proximal convoluted tubule via adenosine A1 receptor. AB - We investigated the role of adenosine A1-receptor in the regulation of basolateral Na(+)-3HCO3- cotransporter in the rabbit proximal convoluted tubule (PCT) microperfused in vitro by monitoring basolateral membrane potential and intracellular pH. FK-453, a highly specific A1 antagonist, inhibited basolateral HCO3- conductance in a concentration-dependent manner (10(-10)-10(-5) M). Other A1 antagonists, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) at 10(-5) M and theophylline at 10(-3) M, also had similar effects. N6-cyclohexyladenosine (CHA) at 10(-7) M attenuated the effect of low concentration (10(-8) M) of FK-453. Either enhancement of the degradation of adenosine by 0.1 U/ml adenosine deaminase (ADA) or inhibition of adenosine release from the cells by 10(-6) M S (4-nitrobenzyl)-6-thioinosine (NBTI) mimicked the effects of A1 antagonists. These observations suggest that endogenous adenosine is released from PCT cells and stimulates Na(+)-3HCO3- cotransporter. Both 10(-4) M 8-(4-chlorophenylthio) adenosine 3',5'-cyclic monophosphate (CPT-cAMP) and 10(-6) M forskolin also inhibited basolateral HCO3- conductance. Both 10(-6) M FK-453 and 10(-4) M CPT cAMP decreased the initial rate as well as the magnitude of intracellular acidification induced by reduction of peritubular HCO3- concentration from 25 to 0 mM. Neither 10(-6) M FK-453 nor 10(-7) M CHA changed intracellular Ca2+ concentration as measured by fura-2 fluorescence. These results indicate that adenosine might stimulate HCO3- exit across the basolateral membrane through Na(+)-3HCO3- cotransporter by decreasing intracellular cAMP via A1-receptor activation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238381 TI - Decreased bone carbonate content in response to metabolic, but not respiratory, acidosis. AB - In vitro cultured neonatal mouse calvariae release calcium and buffer the medium proton concentration in response to a decrease in the medium pH caused by a reduction in bicarbonate concentration ([HCO3-]), a model of metabolic acidosis, but not to an equivalent decrease in pH caused by an increase in the partial pressure of carbon dioxide (PCO2), a model of respiratory acidosis. We have postulated that the medium is in equilibrium with the carbonated apatite in bone. To determine whether bone carbonate is depleted during models of acidosis, we cultured calvariae in control medium (pH approximately 7.4, PCO2 approximately 43, [HCO3-] approximately 26) or in medium in which the pH was equivalently reduced by either a decrease in [HCO3-] (metabolic acidosis, pH approximately 7.1, [HCO3-] approximately 13) or an increase in PCO2 (respiratory acidosis, pH approximately 7.1, PCO2 approximately 86) and determined net calcium flux (JCa) and bone carbonate content. We found that compared with control, after 3, 24, and 48 h there was a decrease in bone carbonate content during metabolic but not during respiratory acidosis. Compared with control, at 3 h JCa increased with both respiratory and metabolic acidosis; however, at 24 and 48 h JCa increased only with metabolic acidosis. JCa was correlated inversely with percent bone carbonate content in control and metabolic acidosis at all time periods studied (r = -0.809, n = 23, P < 0.001). Thus a model of metabolic acidosis appears to increase JCa from bone, perhaps due to the low [HCO3-] inducing bone carbonate dissolution.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238382 TI - Transforming growth factor-beta is a renin secretagogue at picomolar concentrations. AB - Transforming growth factor-beta s (TGF-beta s), a family of peptides, have many actions including modulating cellular growth, differentiation, and influencing steroidogenesis. Because both TGF-beta and renin are present in renal juxtaglomerular cells, we have examined the effects of these peptides on renin secretion using static incubations of rat renal cortical slices. We report here an effect of both TGF-beta 1 and -beta 2 on renin secretion. At low concentrations, both TGF-beta 1 (4 x 10(-12) M) and -beta 2 (8 x 10(-12) M) stimulate basal renin secretion (control, 100 +/- 4%; TGF-beta 1, 123 +/- 4%; TGF beta 2, 124 +/- 5%; both P < 0.02 compared with control). However, at higher concentrations (2 x 10(-10) M), both peptides do not alter basal renin release. Our previous studies show that both prostaglandins and lipoxygenase (LO) products of arachidonic acid play an important dual regulatory role in renin secretion; therefore, we have examined the effects of both cyclooxygenase (CO) and LO inhibition in TGF-beta action.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238383 TI - Endothelin-1 mRNA in glomerular and epithelial cells of kidney. AB - To examine the question of the tubular localization of renal endothelin-1 (ET-1) mRNA, cDNA generated by reverse transcription of isolated rat tubule RNA was amplified by polymerase chain reaction using rat ET-1-specific oligonucleotides. Product identity was determined by restriction enzyme digestion or direct product sequencing. ET-1 mRNA was found to increase in renal tissue in a corticomedullary direction. High levels of ET-1 mRNA were found in dissected glomeruli and in juxtaglomerular cells in short-term primary culture. Among tubule segments, ET-1 mRNA was most abundant in inner medullary collecting ducts (IMCD), but products were also found with cDNA derived from proximal convoluted and straight tubules, thick ascending limbs, and outer medullary collecting ducts. In kidneys of untreated, homozygous Brattleboro rats, the increase of ET-1 mRNA along the corticomedullary axis as well as the preponderance of tubular ET-1 mRNA in IMCD was not observed. Our data show that ET-1 mRNA is present in all nephron segments studied and that its expression may be dependent on the functional state of the kidney. Our results are consistent with the proposal that ET-1 modifies tubular function in an autocrine or paracrine fashion. PMID- 8238384 TI - Phosphatidylcholine-directed phospholipase C: activation by complement C5b-9. AB - In rat membranous nephropathy, complement C5b-9 induces glomerular epithelial cell (GEC) injury and proteinuria. In cultured rat GEC, C5b-9 stimulates a phosphoinositide-directed phospholipase (PL) C and products of PLC downregulate C5b-9-mediated GEC injury. We now report that C5b-9-induced hydrolysis of phosphatidylcholine (PC) provides an additional source of 1,2-diacylglycerol (DAG). PC was labeled in intact GEC by brief incubation with 1-O-[alkyl-3H]2-lyso PC. Assembly of C5b-9 stimulated an increase in PC-derived [3H]DAG (173 +/- 18% control), which was reduced in GEC depleted of protein kinase C (PKC) by prolonged preincubation with phorbol 12-myristate 13-acetate (PMA). Similar to C5b-9, [3H]DAG was released from PC after brief incubation of GEC with Ca2+ ionophore A23187 plus PMA. The increases in [3H]DAG induced by C5b-9 and A23187 plus PMA were paralleled by increases in DAG mass. C5b-9 also increased [3H]phosphatidic acid (PA; 182 +/- 37% control), but there was no significant interconversion of DAG and PA. Thus DAG probably originated via PLC. PC-directed PLC activity was also studied in GEC homogenates by release of [14C]DAG from exogenous 1-palmitoyl-2-[arachidonoyl-14C]PC. PLC activity was present at physiological Ca2+ concentration (200-1,200 nM), and PMA stimulated PLC activity in cell homogenates (in presence of ATP). These results demonstrate directly that PMA stimulates release of DAG from PC and are in keeping with the effect of PMA in [3H]lyso-PC-labeled GEC. Thus GEC contain a PC-directed PLC, whose activity is physiologically regulated and is present at nanomolar Ca2+ concentration. C5b-9 stimulates PC-directed PLC, leading to production of DAG. This DAG might trigger a mechanism for limiting injury during complement attack. PMID- 8238385 TI - Stimulation of alpha 2-adrenergic receptors increases Na(+)-K(+)-ATPase activity in distal convoluted tubule cells. AB - The influence of alpha-adrenoceptor subtypes on Na+ transport in distal convoluted tubules (DCT) has not been examined due to the difficulty of isolating and quantifying responses in this segment. These experiments were designed to test the hypothesis that alpha-adrenergic receptors stimulate Na+ absorption in DCT cells. Norepinephrine and epinephrine increased 22Na+ uptake into immortalized mouse DCT cells by 49 and 55% compared with basal uptake. Selective alpha 2-agonists (guanabenz, clonidine, and B-HT 933) stimulated 22Na+ uptake by 39-45%, but alpha 1-agonists had no effect. alpha 2-Agonist-stimulated 22Na+ uptake was abolished with alpha 2-antagonists (yohimbine, idazoxan). The entry pathways for alpha 2-agonist-stimulated 22Na+ uptake were determined with the NaCl cotransport inhibitor, chlorothiazide (10(-4) M), and the Na+ channel blocker, amiloride (10(-6) M). Agonist-stimulated 22Na+ uptake was inhibited 42 +/- 5 with chlorothiazide and 47 +/- 7% with amiloride. These results suggested that alpha 2-receptors may activate Na(+)-K(+)-adenosinetriphosphatase (Na(+) K(+)-ATPase), resulting in an increased driving force for luminal Na+ entry through both pathways. Ouabain-suppressible 86Rb uptake and intracellular Na+ activity ([Na+]i; measured in single cells on glass cover slips loaded with the fluorescent probe sodium-binding benzofuran isophthalate) were used to measure Na(+)-K(+)-ATPase activity. alpha 2-Agonists significantly increased 86Rb uptake within 30 s. After 2.5 min, epinephrine and B-HT 933 decreased [Na+]i from a control level of 13 +/- 1 to 5 +/- 1 mM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238386 TI - Intracellular Ca2+ and PKC activation do not inhibit Na+ and water transport in rat CCD. AB - Experiments examined the effects of elevation of intracellular calcium concentration ([Ca2+]i) or activation of protein kinase C (PKC) on Na+ and water transport in the rat cortical collecting duct (CCD). We measured the lumen-to bath 22Na+ flux (J1-->b), transepithelial voltage (VT), and water permeability (Pf) in CCD from deoxycorticosterone (DOC)-treated rats. Ionomycin (0.5 and 1 microM) and thapsigargin (1 and 2 microM) were used to increase [Ca2+]i. Phorbol 12-myristate 13-acetate (PMA; 0.3 and 1 microM) and oleoyl-acetyl-glycerol (OAG; 100 microM) were used as activators of PKC. [Ca2+]i was measured in isolated perfused tubules using the fluorescent dye fura 2. When added to the bathing solution, 220 pM arginine vasopressin (AVP) failed to affect [Ca2+]i, whereas 1 microM ionomycin increased [Ca2+]i by 103 +/- 15% and 2 microM thapsigargin increased [Ca2+]i by 24 +/- 4%. In flux studies, neither ionomycin nor thapsigargin affected J1-->b or Pf, although ionomycin caused marked morphological changes. Ionomycin also failed to alter either parameter in tubules from non-DOC-treated rats. Neither 100 microM OAG nor 1 microM PMA affected J1- >b or Pf. OAG at 50 microM had no effect on VT or transepithelial resistance, indicating no inhibition of conductive Na+ transport. We conclude that increased [Ca2+]i and PKC activation do not affect J1--b or Pf in the rat CCD. These findings may account for the sustained increase in J1--b produced in the rat CCD by AVP. PMID- 8238387 TI - Effect of prostaglandin synthesis inhibition on macula densa-stimulated renin secretion. AB - The purpose of the present studies was to evaluate directly the role of prostaglandins in macula densa-mediated renin release. Individual juxtaglomerular apparatus specimens were microdissected from rabbit kidney and perfused with a solution containing either high NaCl (Na+ = 141 meq/l; Cl- = 122 meq/l) or low NaCl (Na+ = 26 meq/l; Cl- = 7 meq/l) concentration. With a step decrease in perfusate NaCl (high to low), renin secretion rate was markedly stimulated (from 15.06 to 63.1 nGU/min, P < 0.01), and the response was almost fully reversible. When specimens were bathed with cyclooxygenase inhibitors flurbiprofen (10(-5) M) or flufenamic acid (10(-4) M), this macula densa-activated increase in renin release was largely or completely abolished (flurbiprofen, 3.5-10.5 nGU/min, not significant; flufenamic acid, 9.0-12.3 nGU/min, not significant). Exposing the macula densa to a step increase in perfusate NaCl concentration (low to high) resulted in a significant and reversible suppression of renin secretion in control specimens, but no significant suppression was seen in specimens treated with flufenamic acid. These data provide direct evidence to support the hypothesis that locally produced prostaglandins may act as a primary mediator of the renin response to macula densa activation. PMID- 8238388 TI - Transforming growth factor-beta 1 regulation of signal transduction in two renal epithelial cell lines. AB - The present studies examine the effect of transforming growth factor-beta 1 (TGF beta 1) on signal transduction pathways in two cultured renal epithelial cell lines. TGF-beta 1 promotes basal and agonist-stimulated adenylate cyclase activity in LLC-PK1 but not MDCK cell membranes. TGF-beta 1 stimulation of LLC PK1 membrane adenylate cyclase activity occurs quickly and can be attenuated by pertussis toxin pretreatment. Both TGF-beta 1 and adenosine 3',5'-cyclic monophosphate (cAMP) exert comparable effects on [3H]thymidine uptake in LLC-PK1 cells, suggesting that TGF-beta 1 regulation of adenylate cyclase activity potentially plays a role in mediating biological responses to TGF-beta 1. The activities of protein kinase C and phospholipase A are not affected by TGF-beta 1 in either LLC-PK1 or MDCK cells. Both TGF-beta 1 and epidermal growth factor (EGF) increase expression and induce the appearance of new forms of the cAMP response element binding protein (CREB) in LLC-PK1 cells. These effects of TGF beta 1 and EGF on CREB appear to be specific since neither TGF-beta 1 nor EGF alters expression of an activating transcription factor in LLC-PK1 cells. The effect of TGF-beta 1 and EGF to alter expression of CREB does not affect CREB binding to its regulatory element in LLC-PK1 cell lysates. These results suggest that some of the biological effects of TGF-beta 1 may be attributed to stimulation of adenylate cyclase activity and cAMP formation as well as to enhanced expression and/or modification of the CREB transcription factor in LLC PK1 cells. PMID- 8238389 TI - Atrial natriuretic peptide initiates Ca2+ transients in isolated renal cortical thick ascending limb cells. AB - The following studies identified and characterized atrial natriuretic peptide (ANP) receptor-mediated Ca2+ transient in cortical thick ascending limb (CTAL) cells. Primary cell cultures were prepared from porcine kidneys by immunodissection, and intracellular Ca2+ concentration ([Ca2+]i) was determined in single cells with microfluorometry. ANP (10(-7) M) and its analogue, C-type natriuretic peptide (CNP, 10(-7) M), elicited Ca2+ transients [104 +/- 6 (basal levels) to 653 +/- 112 nM (stimulated) and from 84 +/- 4 to 209 +/- 18 nM, respectively]. Receptor-mediated [Ca2+]i increase was dose-dependent with a 50% effective concentration (EC50) of approximately 10(-10) M. The increment in [Ca2+]i was due to internal release and influx across the plasma membrane. Prior treatment of ANP or CNP (10(-7)M) did not markedly affect a post application of either ANP or CNP. The truncated analogue of ANP, C-ANP-(4-23), which preferentially binds to clearance receptors, elicited an increase in [Ca2+]i (82 +/- 1 to 427 +/- 41 nM). 8-Bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP) did not alter [Ca2+]i, but pretreatment of CTAL cells with 8-BrcGMP for 30 min before agonist treatment prevented ANP-induced Ca2+ signals [83 +/- 5 (basal) to 88 +/- 5 nM (stimulated)]. These results are evidence for the existence of clearance ANP receptors in CTAL cells that may have biological functions and clearance. The functional responses of these signal interactions may have important consequences on hormone actions with the CTAL. PMID- 8238390 TI - Unilateral carotid-cardiac baroreflex responses in humans. AB - To determine the individual contributions of the right and left carotid sinus baroreflex control of the cardiac interval, we measured heart rate [as expressed by R-R interval (RRI)] and mean arterial pressure (MAP) responses elicited by pulsed trains of neck suction or pressure in 20 healthy men. Pulses were delivered using a customized neck collar device partitioned to allow for stimulation of only one side of the neck. Maximal cardiac gains for right, left, and bilateral baroreflexes were determined from the logistic function of RRI responses to changes in estimated carotid sinus pressure (ECSP). Maximal gains for right (5.44 +/- 1.33 ms/Torr ECSP) and left (4.22 +/- 1.44 ms/Torr ECSP, P > 0.05) carotid-cardiac responses did not differ. However, the bilateral gain (6.89 +/- 1.80 ms/Torr ECSP) was less than the simple sum of the right and left gains (9.67 +/- 2.20 ms/Torr ECSP, P < 0.01). We suggest that the right and left carotid-cardiac reflex responses are quantitatively similar in humans and exhibit an inhibitory interaction when compared with the bilateral response under conditions of supine rest in normotensive subjects. PMID- 8238391 TI - Rapid measurements of diastolic intramyocardial vascular volume. AB - Rapid measurements of coronary vascular volume are necessary for a complete understanding of phasic coronary flow. Because no standard method is capable of making these measurements, we developed a method that uses digital subtraction angiography to image an intravascular contrast agent. The method is capable of measuring vascular volume in 33 ms and can be repeated three times per second. The method was first validated in phantoms. We then used this method to measure coronary vascular volume in the isolated, perfluorochemical-perfused, interventricular canine septum. The speed of the measurements allowed us, for the first time, to directly measure transients in vascular volume following sudden changes in perfusion pressure. At the steady-state and maximal vasodilation, coronary vascular volume varied from 7.5 +/- 1.2 (SE) ml/100 g at a perfusion pressure of 20 mmHg to 12.1 +/- 1.9 ml/100 g at 90 mmHg, which is similar to other reports. After a sudden change of 40 mmHg in perfusion pressure, vascular volume changed with a time constant of 3.2 +/- 0.3 (SE) s. Increasing ventricular wall stretch had no effect on either the steady-state volumes (P = 0.25) or the time constant for volume changes (P = 0.17). The fact that the time constant is longer than the cardiac cycle, yet much shorter than the time necessary to measure vascular volume using other methods, highlights the need for rapid measurements of vascular volume. PMID- 8238392 TI - Left ventricular energetics in closed-chest dogs. AB - Studies of ventricular energetics using the relation between myocardial O2 consumption (MVO2) and pressure-volume area (PVA) have been performed extensively in the isolated heart, but not in the intact animal. We characterized the MVO2 PVA relation and its response to heart rate (HR) in eight closed-chest dogs instrumented with high-fidelity micromanometers, piezoelectric crystals, coronary flow probes, and coronary sinus oximetric catheters. The effect of dobutamine was studied in five dogs. MVO2 is linearly related to PVA with lower MVO2 required for the generation of smaller PVAs. Baseline contractile efficiency (EFF) was 25.6 +/- 2.8%. High pacing rates reduced EFF (25.7 +/- 3.2% at a HR of 107 +/- 3 beats/min vs. 16.3 +/- 2.4% at a HR of 194 +/- 5 beats/min, P < 0.0167) and load independent MVO2 per beat (0.562 +/- 0.119 vs. 0.377 +/- 0.074 J.beat-1 x 100 g LV-1, P < 0.0167) while increasing end-systolic elastance (Ees) (9.4 +/- 1.3 vs. 18.6 +/- 3.1 mmHg/ml, P < 0.0167). Dobutamine administration increased load independent MVO2 per beat (0.392 +/- 0.108 vs. 0.607 +/- 0.083 J.beat-1 x 100 g LV-1, P < 0.05) and contractility (Ees 10.1 +/- 1.5 vs. 32.0 +/- 7.6 mmHg/ml, P < 0.05) without changing EFF (28.8 +/- 3.8 vs. 30.3 +/- 3.8%, P = NS). Thus the intact animal displays loss of EFF at high heart rates but maintains EFF during dobutamine stimulation. Both interventions increased load-independent MVO2 per minute, indicating increased O2 requirements for excitation-contraction coupling. PMID- 8238393 TI - Pulmonary endothelial nitric oxide production is developmentally regulated in the fetus and newborn. AB - To define the role of endothelial nitric oxide (NO) in developmental changes in pulmonary vascular resistance and oxygen responsiveness, we determined the ontogeny of endothelial NO production and of oxygen modulation of that process in pulmonary arteries from fetal and newborn lambs. NO production was assessed by measuring endothelium-dependent arterial guanosine 3',5'-cyclic monophosphate synthesis. Basal NO rose two-fold from late gestation to 1 wk of age and another 1.6-fold from 1 to 4 wk. Acetylcholine-stimulated NO also increased 1.6-fold from 1 to 4 wk. The maturational rise in NO was evident at high Po2 in vitro, and it was not modified by L-arginine. This suggests that the developmental increase may alternatively involve enhanced calcium-calmodulin-mediated mechanisms, increased expression of NO synthase, or greater availability of required cofactor(s). With an acute decline in Po2 in vitro from 680 to 150 or 40 mmHg, there was 50-88% attenuation of basal and acetylcholine-stimulated NO late in the third trimester and in the newborn but not early in the third trimester. Parallel studies of mesenteric endothelium revealed postnatal increases in basal and stimulated NO but no decline in NO at lower Po2. Ontogenic changes in endothelial NO production and in oxygen modulation of that process may be involved in the maturational decrease in vascular resistance and the development of oxygen responsiveness in the pulmonary circulation. PMID- 8238394 TI - Purine metabolic pathways in rat hindlimb perfusion model during ischemia and reperfusion. AB - The perfused rat hindlimb preparation was used with a blood cell-free perfusate to investigate alterations in the purine nucleotide metabolism, flow rate, perfusion pressure, and venous excretion in response to ischemia and ischemia followed by reperfusion in skeletal muscle. The development of a physical hindrance during postischemic reperfusion, indicated by an increase in reperfusion pressure and a decrease in flow rate, coincided with a 90% decrease in phosphocreatine and a 50-70% reduction in total adenine nucleotide pool. The reflow impairment could not be explained by blood cell plugging of the capillaries. Washout of several metabolites was demonstrated during reperfusion. Hypoxanthine accumulated intracellularly during ischemia, and a substantial amount of uric acid was excreted into the venous effluent during reperfusion. The experimental data were fitted into a computer simulation model of the purine pathways. The model indicated that AMP deaminase was the predominant enzymatic pathway for the AMP degradation. It was demonstrated that ATP preferably accumulated as inosine-5'-monophosphate during ischemia and that xanthine oxidase was undetectable in skeletal muscle tissue homogenates. However, vascular endothelial cell xanthine oxidase activity responsible for a free radical-induced reperfusion injury could not be excluded. PMID- 8238395 TI - Possible paracrine facilitation of ventricular growth in oculo by atrial tissue. AB - Load-independent growth controls were investigated in embryonic atria and ventricles cultured in the anterior eye chamber of adult rats. At embryonic day 12, the mass of atria was half that of ventricles (0.11 vs. 0.24 mg), whereas after 8 wk in oculo, atrial grafts were threefold larger than ventricular grafts (2.26 +/- 0.36 vs. 0.69 +/- 0.16 mg). To examine possible paracrine communication between atria and ventricles, two grafts were cocultured in the same eye chamber. Atrial grafts cocultured with either another atrial graft or a ventricular graft did not differ in projected area or mass from atrial grafts cultured alone. Ventricular grafts cocultured with another ventricular graft did not differ in projected area or mass from ventricular grafts cultured alone. However, ventricular grafts cocultured with an atrial graft grew larger than single ventricular grafts. At 1 wk in oculo, DNA synthesis (bromodeoxyuridine incorporation) was higher in ventricular grafts cocultured with atrial grafts than in single ventricular grafts (6.84 +/- 0.88 vs. 4.40 +/- 0.10%). Thus atria may stimulate growth, including DNA synthesis, in developing ventricles via a paracrine mechanism. PMID- 8238396 TI - Inwardly rectifying potassium current in rat fetal and neonatal ventricular cardiomyocytes. AB - Whole cell and single-channel inwardly rectifying potassium currents (IK1) of freshly isolated single fetal (12 and 18 days) and neonatal (1, 5, and 10 days) rat ventricular myocytes were recorded using patch-clamp techniques. Whole cell IK1 was elicited by hyperpolarizing test pulses from a holding potential of -40 mV. IK1 densities increased markedly during heart development between fetal day 12 and until neonatal day 5; there was no further increase on neonatal day 10. Cell-attached patch recordings of single IK1 channels were employed with 150 mM K+ both in the pipette and bath solutions. Large-conductance (31 pS) K+ channels with short openings were observed both in fetal day 18 and neonatal day 5 but not in fetal day 12 cells. Only small-conductance (11 pS) channels with long openings were observed in approximately 10% of the cells on fetal day 12. The open probability of the large-conductance channels was 2.6-fold greater on neonatal day 5 than on fetal day 18. In conclusion, the increase in the IK1 density during observed heart development may be primarily due to an increase in the open probability of the IK1 channels, rather than to an increase in their number. We suggest that the small-conductance channel infrequently observed on fetal day 12 could be an immature type of IK1 channel, which disappears during development. PMID- 8238397 TI - Mechanical determinants of coronary blood flow during dynamic alterations in myocardial contractility. AB - Recently it has been proposed that the decrease in coronary blood flow (CBF) resulting from cardiac contraction referred to as systolic flow impediment (SFI) is dependent on the level of left ventricular elastance (Ees). The average rate of LV relaxation (Ravg) has been shown to be major determinant of diastolic flow development (DFD). We tested these hypotheses using the unique hemodynamic condition of pulsus alternans (PA) where end-systolic LV pressure and instantaneous Ees vary on beat-to-beat basis. In six mongrel dogs instrumented with LV and aortic manometers, ultrasonic dimension crystals, and Doppler coronary flow probes we measured phasic CBF and Ees during PA and control conditions. Maximal pressure development over time (dP/dtmax) and SFI were significantly different between weak (WB) and strong beats (SB) as were Ravg and DFD. Minimum CBF (Qmin) was not different between SB and WB; however, Qmin and peak Ees occurred nearly simultaneously in the WB. Qmin occurred much earlier than peak Ees in the strong and control beats. Plots of instantaneous LV elastance and CBF showed that for control beats and for the strong beats of PA CBF was similar during systole and diastole, suggesting elastance is a unique determinant of CBF. This was quantified as CBF at the time in either systole or diastole when elastance was half-maximal for that beat (E50). During the WB of PA, however, CBF at E50 was significantly higher during systole than during diastole. We conclude that while SFI and DFD are highly dependent on the dP/dt and Ravg, Ees is not a unique determinant of CBF under all conditions. PMID- 8238398 TI - Exhaustion of Frank-Starling mechanism in conscious dogs with heart failure. AB - The goal of this study was to elucidate the ability of the left ventricle to accommodate an increase in preload (Frank-Starling mechanism) in the presence of congestive heart failure (CHF) but in the absence of the complicating effects of hypertrophy and fibrosis. To accomplish this, the effects of volume loading were examined in eight conscious dogs during the control state and after 3 wk of right ventricular pacing (240 beats/min). CHF increased heart rate (by 16 +/- 5 from 92 +/- 5 beats/min), left ventricular (LV) end-diastolic pressure (by 17 +/- 2 from 10 +/- 1 mmHg), and LV end-diastolic volume (EDV; by 23 +/- 4 from 57 +/- 3 ml). Despite reduced LV ejection fraction (from 54 +/- 3 to 31 +/- 3%), there was no significant change in cardiac output (2.5 +/- 0.3 l/min) compared with control (2.7 +/- 0.2 l/min). Stroke volume was preserved (control 19 +/- 2 ml; CHF 18 +/- 2 ml) at a constant heart rate by a shift to the right in the relationship between LV stroke volume and EDV, indicating the importance of chronic ventricular dilatation in maintaining pump performance. In the control state, acute volume load increased LV EDV (by 17 +/- 2 ml) and stroke volume (by 11 +/- 2 ml), whereas in CHF it did not increase LV EDV or stroke volume. Scanning electron microscopy revealed areas of reduced collagen weave pattern surrounding myofibers. Myocyte cross-sectional area by transmission electron microscopy was significantly reduced, and there were multiple electron-dense expansions of the Z lines with disruption of the normal lateral sarcomere alignment. These morphological findings suggest that chronic ventricular dilatation utilized in CHF results from myocyte stretch and morphological intracellular rearrangement. Furthermore, the failing heart cannot further augment stroke volume by acutely increasing EDV in CHF, suggesting that the Frank-Starling reserve is essentially exhausted. PMID- 8238399 TI - Guanethidine chemical sympathectomy: spinal cord and sciatic nerve blood flow. AB - The spinal cord vasculature is innervated by noradrenergic nerve fibers, the role of which in the regulation of regional spinal cord blood flow (RSCBF) is presently unclear. We used the distribution of [14C]butanol to simultaneously measure RSCBF at seven cord levels and the regional blood flow in sciatic nerve (NBF), truncal skin, and biceps femoris muscle. The subjects were control rats and rats that had been given parenteral guanethidine sulfate for 5 wk to induce selective postganglionic "chemical sympathectomy." Flows were measured under basal conditions (group I) and immediately after an arterial hemorrhage (group II). The results indicate that RSCBF was unchanged from control after guanethidine administration in both groups; however, NBF was elevated after guanethidine by 47% in group I and by 41% in group II. We conclude that in the spinal cord as in the brain, sympathetic inflow does not appear to have an important role in the regulation of regional blood flow. Sympathetic inflow appears to partly regulate NBF, however, probably by varying vascular tone. PMID- 8238400 TI - Effect of muscle length on the in vitro comparison of femoral arteries before and after endotoxin shock. AB - Control and endotoxin-treated femoral arteries were compared in vitro for the effect of muscle length. Rats were anesthetized with pentobarbital, and endotoxin (6 mg/kg) was infused for 1 h. A control ring before endotoxin treatment and a ring after endotoxin treatment (blood pressure = 40 mmHg) were excised from the contralateral artery for length-tension and dose-response experiments with phenylephrine. The initial length for resting tension (Li) was shorter for endotoxic rings (1.23 +/- 0.01 vs. 1.41 +/- 0.02 mm in control), but the length of maximum active tension (Lmax) was the same. In length-tension experiments the values for active tension (6.36 +/- 0.61 vs. 4.06 +/- 0.60 x 10(3) dyn/cm), preload at Lmax (1,333 +/- 204 vs. 733 +/- 146 mg), and passive stiffness were increased after endotoxin. In dose-response experiments at the same preload, the endotoxic rings had a lower active tension (3.28 +/- 0.28 vs. 6.55 +/- 0.27 x 10(3) dyn/cm) but the same sensitivity. At Lmax, active tension (12.45 +/- 0.48 vs. 5.01 +/- 0.89 x 10(3) dyn/cm in control vessels) and sensitivity (half maximum effective dose = 0.68 +/- 0.8 x 10(-6) vs. 1.39 +/- 0.29 x 10(-6) M in control vessels) were greater for endotoxic rings. These experiments show that phenylephrine sensitivity and active tension in the rat femoral artery are increased by endotoxin shock, and the importance of muscle length is implied. PMID- 8238401 TI - Regulation of Na-K-ATPase gene expression by aldosterone in vascular smooth muscle cells. AB - Na-K-adenosinetriphosphatase (ATPase) activity profoundly influences vascular cell excitability, contractility, and volume regulation. The recent finding of mineralocorticoid hormone receptors in vascular tissue suggests the possibility that Na-K-ATPase gene expression in vascular tissue is regulated by the mineralocorticoid aldosterone. In this study, we investigated Na-K-ATPase gene expression by aldosterone in cultured rat vascular smooth muscle cells (VSMC). Na K-ATPase alpha 1- and beta 1-isoform mRNAs, but not alpha 2- and alpha 3-isoform mRNAs, were expressed in cultured rat VSMC. Aldosterone caused a 2.3-fold increase in the alpha 1 mRNA and a 4.7-fold increase in the beta 1 mRNA accumulation with peak elevations at 24 and 6 h, respectively. Aldosterone induced the alpha 1 mRNA expression at physiological concentrations (half-maximum effective concentration = 2-3 nM), consistent with the binding of aldosterone to mineralocorticoid hormone receptors. The augmented alpha 1 mRNA expression by aldosterone was associated with a twofold increase in the alpha 1-subunit protein accumulation. Pretreatment of VSMC with cycloheximide caused a 10-fold increase in the alpha 1 mRNA expression, and the aldosterone-mediated alpha 1 mRNA accumulation was not observed in the presence of cycloheximide. Transfection experiments with the luciferase reporter gene revealed that aldosterone response sequences are located within the 5'-flanking regions of the alpha 1-isoform gene. These data demonstrate that the mineralocorticoid aldosterone directly stimulates Na-K-ATPase gene expression and protein accumulation in VSMC. PMID- 8238402 TI - Manganese-superoxide dismutase in endothelial cells: localization and mechanism of induction. AB - Mechanisms of Mn-superoxide dismutase (Mn-SOD) expression in human umbilical endothelial cells were investigated by Northern blot analysis, enzyme-linked immunosorbent assay, and immunoelectron microscopy. The Mn-SOD in human endothelial cells was markedly induced by the cytokines tumor necrosis factor (TNF), interleukin-1, and lipopolysaccharide as well as by phorbol esters [12-O tetradecanoylphorbol 13-acetate (TPA)]. The induction was partially blocked by dexamethasone and 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine, a potent inhibitor of protein kinase C (PKC). In endothelial cells in which PKC had been desensitized to TPA by pretreatment for 24 h, addition of TNF caused overexpression of Mn-SOD. These facts suggested that at least two separate signal transducing pathways are involved in expression of the Mn-SOD gene. Immunoelectron-microscopic studies showed that Mn-SOD was localized to the mitochondrial matrix of the capillary vascular endothelial cells of cardiac tissues and cultured endothelial cells. Mn-SOD, which is normally abundant in endothelial cells relative to other cell types, may play an important protective role against stresses such as ischemia and inflammation. PMID- 8238403 TI - Attenuation of postexertional hypotension by cardiac afferent blockade. AB - Naloxone eliminates postexertional hypotension (PEH) in human and animal models. The effect of naloxone on sympathetic activity during hemorrhage and generation of arterial baroreflex function curves can be stimulated by blockade of cardiac afferent receptors. We tested the hypothesis that cardiac afferent blockade would eliminate PEH in eight spontaneously hypertensive rats (SHR). Rats were instrumented with a Silastic-tipped catheter inserted into the pericardial space. Four weeks later, a Teflon catheter was placed in the descending aorta via the left common carotid artery for measurement of mean arterial pressure (MAP) and heart rate (HR). MAP and HR were examined before (preexercise) and after (postexercise) a single bout of dynamic treadmill exercise (9-12.0 m/min, 10-18% grade for 30-40 min) under three experimental conditions: control, cardiac efferent blockade, and combined cardiac efferent and afferent blockade. MAP significantly decreased (29 +/- 5 and 25.6 +/- 4 mmHg) in the control and cardiac efferent blockade conditions after exercise. However, when cardiac afferents were blocked, the hypotensive response to mild dynamic exercise was significantly attenuated (-6 +/- 3 mmHg). Thus blockade of cardiac afferents eliminated PEH in the SHR. These data suggest that inhibitory influence of cardiac afferents on the circulation may be enhanced after exercise. PMID- 8238404 TI - Acute exercise enhances nitric oxide modulation of vascular response to phenylephrine. AB - The influence of the release of endothelium-derived nitric oxide (NO) on the vasoconstrictor response to phenylephrine (PE) was evaluated before and after a single bout of dynamic exercise. Each rat ran on a motor-driven treadmill at 12 18 m/min, 10-18% grade until exhaustion (avg time 45 min). Sprague-Dawley rats (n = 6) were instrumented with a Doppler ultrasonic flow probe around the right common iliac artery. Just distal to the flow probe, a catheter was placed into the right iliac artery for local infusions. A Teflon catheter was placed in the descending aorta to measure mean arterial blood pressure (MAP) and heart rate (HR). PE (0.005-0.075 microgram/kg) and NO inhibitor N omega-nitro-L-arginine methyl ester hydrochloride (L-NAME, 0.2-0.25 mg/kg) were injected into the functionally isolated hindlimb. HR and MAP were not altered by any of the injections because we selected doses below those which elicited systemic responses. Dose-response curves to PE were generated in the control and postexercise condition, with and without the NO synthase inhibitor L-NAME. Exercise significantly attenuated the maximal vasoconstrictor response to PE (45.6 +/- 1.6%). L-NAME enhanced the maximal vasoconstrictor response to PE 49.8 +/- 4.5% in the control condition and 121.4 +/- 5.9% in the postexercise conditions. Thus, although NO inhibition enhanced the vasoconstrictor response to PE in the control and postexercise conditions, the enhanced vasoconstrictor response to PE after L-NAME was significantly greater in the postexercise condition. Results suggest that NO contributes to the exercise induced attenuation of alpha 1-adrenergic receptor stimulation. PMID- 8238405 TI - Reduced Na(+)-K+ pump activity in diabetic rabbit carotid artery: reversal by aldose reductase inhibition. AB - This study addresses the question of whether a decrease in basal Na+ pump [Na(+) K(+)-adenosinetriphosphatase (ATPase)] activity occurs in the carotid artery of an alloxan-diabetic rabbit and, if so, whether it is associated with altered 86Rb+ uptake and contractile response to ouabain and K(+)-free solution. Ouabain sensitive 86Rb+ uptake, an index of Na+ pump activity, was diminished approximately 50% in carotid arteries from diabetic rabbits. Concurrent with this, contractions induced by incubating the carotid arteries in a K(+)-free solution (in the absence of phentolamine) were significantly larger in the diabetic group. Readdition of K+ (1 mM) to arteries contracted with the K(+)-free solution caused relaxations that were slower to occur and of lesser magnitude in diabetic than in control rabbits. In contrast to the contractions caused by the K(+)-free medium, contractions caused by incubation with ouabain (1 mM) in the presence of phentolamine were significantly smaller in the diabetic group. Treatment of diabetic rabbits with an aldose reductase inhibitor, zopolrestat, at both high and low doses restored the alterations in vascular reactivity toward normal. The results indicate that the Na+ pump activity is diminished in the carotid artery of diabetic rabbit, and this is associated with abnormal vascular responsiveness and increased polyol pathway flux. PMID- 8238406 TI - Regulation of cardiac mitochondrial calcium by average extramitochondrial calcium. AB - A system to perifuse isolated rat heart mitochondria was designed to study the relationship between mitochondrial matrix free Ca2+ and extramitochondrial free Ca2+ under conditions in which the latter concentration could oscillate over a range typical of that expected in vivo. We tested the hypothesis that the level of intramitochondrial Ca2+ responds to the average extramitochondrial Ca2+ in the heart. Mitochondria were immobilized within an optical chamber for measurement of endogenous NAD(P)H and fura 2 fluorescence. NAD(P)H increased significantly on provision of substrates and decreased reversibly in the presence of ADP, indicating maintenance intact coupled respiration by this preparation. Matrix free Ca2+ was measured using fura 2-loaded mitochondria and, in parallel experiments, media free Ca2+ was measured with fura 2 in the absence of mitochondria. Oscillation of extramitochondrial Ca2+ from < 0.1 microM to approximately 2 microM at frequencies of 0.5, 1.0, and 1.25 cycles/s produced steady-state levels of matrix Ca2+ that were independent of frequency but proportional to the average media free Ca2+ concentration. Matrix Ca2+ increased to a steady state on an increase in the extramitochondrial average Ca2+ concentration with a half-time (t1/2) of approximately 2 min at 22 degrees C. Oscillation of mitochondrial Ca2+ was not observed under any conditions tested. The data are taken to indicate that in vivo, the concentration of mitochondrial matrix free Ca2+ is a steady state that is proportional to the average extramitochondrial Ca2+ concentration and that changes in the latter represent a mechanism of signal transduction from the cytosol to the mitochondrial matrix. PMID- 8238407 TI - Kallikrein release by vascular tissue. AB - Vascular tissue contains kallikrein and kallikrein mRNA, suggesting a vascular kallikrein-kinin system. We questioned whether 1) kallikrein concentration varies among large and small vessels; 2) kallikrein is released by vascular tissue; and 3) blocking protein synthesis inhibits release, suggesting de novo synthesis. Using rat vascular rings and isolated-perfused hindquarters, we examined kallikrein in the bath and perfusate. Active kallikrein was higher in tail arteries than the aorta (P < 0.001); tail veins had six times more kininogenase than the vena cava (P < 0.001). Total kallikrein showed a similar pattern, being highest in tail vessels. Arterial rings released active and total kallikrein. After 1, 2, and 3 h incubation, cumulative release was as follows: active, 90 +/- 13, 201 +/- 25, and 311 +/- 41 pg.h-1 x mg tissue-1; total, 170 +/- 14, 366 +/- 24, and 537 +/- 40 pg.h-1 x mg tissue-1, indicating constant release up to > or = 3 h. In contrast, lactic dehydrogenase fell from 6.7 +/- 2.5 to 2.5 +/- 0.4 U.h-1 x mg tissue-1. Total kallikrein in the rings was 302 +/- 51 pg bradykinin/mg wt tissue before 3 h and 298 +/- 68 afterward. Kallikrein released by the hindquarters after 3 h was as follows: active, 6.2 +/- 2.8 ng bradykinin.min-1 x kg.body wt-1; total, 85.2 +/- 17 ng bradykinin.min-1 x kg body wt-1. Puromycin pretreatment (10 mg ip) reduced total perfusate kallikrein from 105 +/- 19 to 8.5 +/- 3.6 (P < 0.005).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238408 TI - Effect of tetanic myocardial contraction on coronary pressure-flow relationships. AB - Cardiac contraction causes a decrease in coronary flow. Despite many studies, it is still not clear what mechanism or mechanisms are responsible for this flow decrease. The phasic nature of myocardial contraction and the complexities intrinsic to intact heart preparations make it difficult to elucidate the mechanisms. We therefore studied coronary pressure-flow relationships during steady-state (tetanic) contractions in the maximally vasodilated isolated canine interventricular septum to see whether waterfall-type behavior is present. Using ryanodine and electrical stimulation allowed the production of reproducible and reversible tetani. This preparation minimizes the difficulties associated with transmural variations and also the effects of intramyocardial capacitance. Two separate protocols were performed to delineate the pressure-flow relationships in the passive and tetanized states. The first compared diastolic and tetanized pressure-flow relationships. In the second protocol, 2,3-butanedione monoxime was added to obtain an intermediate contractile level, thus allowing the comparison of two contractile states. Both the diastolic and tetanized pressure-flow relationships were curvilinear in the low-pressure range. Linear and nonlinear fits to the data showed that the primary effect of contraction was a shift of the pressure-flow relationships to higher pressures at a given flow. This effect was graded by the level of contractility and was independent of developed stress. Although other mechanisms may also be operative, these results support the presence of waterfall behavior in the coronary vascular bed. PMID- 8238409 TI - Is rapid rise in vascular conductance at onset of dynamic exercise due to muscle pump? AB - We tested the hypothesis that rapid increases in muscle blood flow and vascular conductance (C) at onset of dynamic exercise are caused by the muscle pump. We measured arterial (AP) and central venous pressure (CVP) in nine awake dogs, eight with atrioventricular block, pacemakers, and ascending aortic flow probes for control of cardiac output (CO) (2 also had terminal aortic flow probes). One dog had only an iliac artery probe. At exercise onset (0 and 10% grade, 4 mph) C and CVP rose to early plateaus, and AP reached a nadir, all in 2-5 s. At 20% grade and 4 mph, C increased continuously after its initial sudden rise. Timing and magnitude of initial change in conductance (delta C) were independent of CO, AP, work rate (change in grade at constant speed), or autonomic function (blocked by hexamethonium). Speed of initial delta C and its independence from work rate and blood flow ruled out metabolic vasodilation as its cause; insensitivity to AP and autonomic blockade ruled out myogenic relaxation and sympathetic vasodilation as causes of sudden delta C. Sensitivity to contraction frequency (not work per se) implicates the muscle pump. When reflexes were blocked, a large secondary rise in C, presumably caused by metabolic vasodilation, began after 10 s of mild exercise. When reflexes were intact in mild exercise, C was lowered below its initial plateau by sympathetic vasoconstriction, which partially raised AP from its nadir toward its preexercise level. Our conclusion is that dynamic exercise has a large rapid effect on C that is not explained by known neural, metabolic, myogenic, or hydrostatic influences.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238410 TI - Evidence for sensing and integration of biological signals by the capillary network. AB - The aim of this study was to explore the phenomenon first described by Dietrich (Microvasc. Res. 38: 125-135, 1989) in which a local application of norepinephrine (NE) on a capillary can temporarily reduce flow via constriction of the feeding arteriole. Our objectives were to show that this phenomenon of remote response is not limited to vasoconstriction, can be elicited by materials other than NE, shows stimulus-strength dependency, and can be integrated within the capillary network. We used an intravital micropharmacological approach to iontophoretically apply (in mM concentrations in the pipette) NE, acetylcholine (ACh), 5'-N-ethylcarboxamidoadenosine (NECA, adenosine analogue), K+, and H+ on capillaries of the frog sartorius muscle in situ. Responses were measured in terms of changes in velocity of red blood cells (VRBC) in capillaries or in terms of changes in arteriolar diameter. ACh (3 mM) caused significant increases in diameter (from 34 to 37 microns) and in VRBC (from 250 to 340 microns/s, i.e., 36%). NE (3 mM) reduced VRBC by 16%. The magnitude of ACh and NE velocity responses increased with increasing pipette concentration and with increasing iontophoretic current. The ACh response was blocked by a local pretreatment with atropine. NECA, K+, and H+ caused 20-40% increases in VRBC. Dual application of NE on two capillaries fed by the same arteriole resulted in a greater VRBC reduction than for single NE application. Dual application of NE and ACh significantly attenuated the ACh response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238411 TI - Local cerebral blood flow and glucose utilization after blood exchange with a hemoglobin-based O2 carrier in conscious rats. AB - The effects of a blood exchange on cerebral blood flow and glucose utilization were studied. A near to total blood exchange (hematocrit < 3%) was achieved in conscious rats by isovolemic hemodilution. Ultrapurified, polymerized, bovine hemoglobin (UPBHB) served as a blood substitute. Local cerebral blood flow (LCBF) and local cerebral glucose utilization (LCGU) were measured in 34 brain structures of conscious rats by means of the ido[14C]antipyrine and the 2-[14C] deoxy-D-glucose methods. A group of rats without blood exchange served as control. After blood exchange LCBF increased from 36 to 126% in the different brain structures resulting in a nearly doubled mean cerebral blood flow (+82%). LCGU increased only moderately by 0-24%. Significant increases in LCGU were observed in 16 brain structures. Mean cerebral glucose utilization slightly increased (+14%). The relationship between LCGU and LCBF was found to be tight both in the control group (r = 0.95) as well as after blood replacement (r = 0.94), although it was reset to a higher overall LCBF-to-LCGU ratio. The profound increases in LCBF observed after blood exchange, which were not paralleled by comparable increases in LCGU, might be explained by a reduction of blood viscosity after blood exchange. Additional effects of blood exchange observed in the present study were an increase of mean arterial blood pressure and a decline of heart rate. The results indicate that replacement of blood with the hemoglobin based oxygen carrier UPBHB appears to meet the cerebral circulatory and metabolic demands of the brain tissue. PMID- 8238412 TI - Adenosine deaminase inhibitors attenuate ischemic injury and preserve energy balance in isolated guinea pig heart. AB - We investigated the effect of the adenosine deaminase inhibitors erythro-9-(2 hydroxy-3-nonyl)adenine (EHNA) and coformycin on high-energy phosphate metabolism, tissue nucleotides and nucleosides, and recovery of contractile function in isolated, perfused guinea pig hearts. EHNA and coformycin (10 microM) improved postischemic recovery of contractile function approximately 85% and enhanced coronary flow rate in reperfused tissue approximately 40%. The protective effect of EHNA on recovery of contractile function was concentration dependent. Although adenosine (10 microM) increased coronary flow rate on reperfusion approximately twofold over vehicle, it failed to improve postischemic recovery of contractile function. EHNA and coformycin preserved cardiac ATP levels and increased endogenous tissue adenosine during ischemia. During reperfusion, these agents enhanced recovery of high-energy phosphates approximately twofold and potentiated adenosine release into the perfusate with concentration dependency. Furthermore, EHNA and coformycin reduced the extent of myocardial ischemia-reperfusion injury, as indicated by the approximately 55% reduction in creatine phosphokinase release. We conclude that inhibitors of adenosine deaminase attenuate myocardial ischemic injury and improve postischemic recovery of contractile function and metabolism through endogenous myocardial adenosine enhancement and ATP preservation. PMID- 8238413 TI - Rate-dependent functional properties of retrograde atrioventricular nodal conduction in experimental animals. AB - While rate-dependent atrioventricular (AV) nodal functional properties play a major role in determining antegrade AV nodal conduction, their existence and characteristics have not been assessed during retrograde AV nodal impulse propagation. Pacing protocols were used to study selectively AV nodal recovery, facilitation, and fatigue in 6 isolated, superfused rabbit AV nodal preparations and in 11 morphine-chloralose anesthetized dogs. All three properties were identifiable during retrograde AV nodal activation in rabbits. Retrograde recovery and fatigue were clearly demonstrated in dogs, but facilitation could not be evaluated because of echo beats during retrograde premature stimulation. Functional properties were qualitatively similar during retrograde and antegrade propagation; however, important quantitative differences were noted. The time constant for recovery from activation was significantly greater in the retrograde [rabbits, 69 +/- 8 (SE) ms; dogs, 93 +/- 11 ms] compared with the antegrade direction (rabbits, 50 +/- 5 ms; dogs, 58 +/- 4 ms; P < 0.05 vs. retrograde for each species). The magnitude of fatigue resulting from sustained increases in rate was also substantially greater in the retrograde direction in both rabbits (17 +/- 2 vs. 10 +/- 1 ms antegrade, P = 0.01) and dogs (20 +/- 4 vs. 6 +/- 1 ms antegrade, P < 0.01). Conduction time and refractory period were both greater in the retrograde compared with antegrade direction, and directional differences in conduction properties were magnified as activation rate increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238414 TI - Pressure-independent baroreflex resetting produced by chronic infusion of angiotensin II in rabbits. AB - The purpose of this study was to test the hypothesis that chronic infusion of angiotensin II (ANG II) in rabbits shifts or resets baroreflex control of heart rate to a higher pressure level via a mechanism that is independent of the hypertension that is produced. The baroreflex relationship between arterial pressure and heart rate was assessed by first infusing progressively increasing doses of nitroprusside (3, 6, 12, 24, and 48 micrograms.kg-1 x min-1) to lower pressure and then increasing doses of phenylephrine (0.5, 1, 2, 4, and 8 micrograms.kg-1 x min-1) to raise pressure. Two weeks of intravenous ANG II infusion (20 ng.kg-1 x min-1) increased plasma ANG II levels from 9 +/- 1 to 146 +/- 24 pg/ml (P < 0.05), increased arterial pressure from 62 +/- 2 to 95 +/- 2 mmHg (P < 0.05), and transiently increased heart rate. The baroreflex was shifted to a higher pressure level after 30 min and 1, 3, 7, 9, and 14 days of ANG II infusion. Thirty minutes after the ANG II infusion on days 1, 7, and 14 was stopped, arterial pressure decreased, and the baroreflex shifted back to control, indicating that ANG II was required for the resetting that was produced. However, when the ANG II infusion was continued and arterial pressure was instead reduced for 30 min by infusing nitroprusside on days 3 and 9, the baroreflex relationship between arterial pressure and heart rate remained positioned at a higher pressure level.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238415 TI - Mechanisms of isoproterenol-induced atrial natriuretic peptide release from superfused rabbit atria. AB - The mechanisms for isoproterenol-induced atrial natriuretic peptide (ANP) release were studied in superfused rabbit sliced right atria. Addition of 1 microM norepinephrine to this preparation induced a significant monophasic twofold rise in ANP release. This effect was abolished by 1 microM propranolol and mimicked by 1 microM isoproterenol. Furthermore, addition of 200 microM 3-isobutyl-1 methylxanthine (IBMX) to the superfusing medium potentiated isoproterenol effect 31%. In addition, superfusion of slices with 0.5 mM N6,2-O-dibutyryladenosine 3',5'-cyclic monophosphate [(Bu)-2cAMP] enhanced ANP release in the same manner as the beta-agonist. After isoproterenol stimulation, adenosine 3',5'-cyclic monophosphate (cAMP) concentration in effluents increased significantly. ANP secretory response to isoproterenol was unaffected by extracellular calcium concentration or 1 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N' tetraacetic acid (EGTA). Finally, 10 microM indomethacin significantly reduced isoproterenol-stimulated ANP release. It is concluded that 1) norepinephrine induced ANP release is mediated by its beta-agonist activity, 2) isoproterenol stimulated release appears to be mediated by cAMP, 3) isoproterenol effect does not require extracellular calcium, and 4) prostaglandins may be involved in this beta-adrenergic effect. PMID- 8238416 TI - Adenosine contributes to hypoxia-induced vasodilation through ATP-sensitive K+ channel activation. AB - the relative contribution of adenosine and ATP-sensitive potassium (KATP) channels in hypoxia-induced vasodilation was studied in isolated, saline-perfused rabbit hearts under constant flow conditions. Adenosine infusion induced a dose dependent reduction in coronary perfusion pressure. Inhibition of KATP-channels with glibenclamide (0.3 microM) significantly reduced the dilator response to adenosine (-26 +/- 6 ys. -6 +/- 2% at 1 microM), whereas those of 1 microM acetylcholine or 10 microM papaverine were unaffected. The dilator responses to the A1-receptor selective agonist, N6-cyclopentyladenosine (CPA), and the mixed A1- and A2-receptor agonist, 5'-(N-ethylcarboxamido) adenosine (NECA), were comparable to that of adenosine. Treatment with glibenclamide reduced partially but significantly the NECA-induced vasodilation (-28 +/- 4 vs. -17 +/- 4%) and changed the CPA dilator response to a significant vasoconstriction. Both the nonselective adenosine-receptor antagonist, 8-phenyltheophylline (10 microM), and glibenclamide blocked the dilator response to adenosine and significantly reduced the hypoxia-induced vasodilation to the same extent. In conclusion, we suggest that the activation of KATP channels plays a major role in hypoxia-induced vasodilation. A major part of this activation results from the action of adenosine on receptors closely related to the A1 type. PMID- 8238417 TI - Importance of venular flow in control of arteriolar diameter in hamster cremaster muscle. AB - These experiments tested the hypothesis that an intact venular flow is important for the control of upstream arteriolar diameter during an increase in metabolism. A silicone stopcock grease dam was placed across the distal portion of the cremaster muscle to localize treatment of the metabolic stimulator 2,4 dinitrophenol (DNP). Thus only the distal area would have an increased metabolic rate, with the proximal area of the cremaster having a normal metabolic rate. During DNP treatment, the first-order venule, approximately 5 mm proximal to the Silastic dam, was occluded to prevent the transport of metabolites. DNP treatment (10 mM) resulted in a significant increase in the arteriolar diameter from 75 +/- 3 to 90 +/- 4 microns (n = 7, P < 0.05), 12.1 +/- 0.3 mm upstream from the distal region. After venular occlusion, arteriolar diameter decreased to 78 +/- 3 microns (P < 0.05). As an additional test of our hypothesis we altered the experimental sequence. DNP treatment during venular occlusion did not affect arteriolar diameter, but after release of the occlusion there was a significant increase in arteriolar diameter from 78 +/- 3 to 91 +/- 4 microns (P < 0.05). These results suggest that an intact venular flow is necessary for control of arteriolar diameter during an increased metabolic rate caused by DNP treatment, providing evidence for the significance of the venular-arteriolar diffusion of vasoactive metabolites. PMID- 8238418 TI - Sodium current in isolated human ventricular myocytes. AB - Although fast sodium current (INa) plays a major role in the generation and conduction of the cardiac impulse, the electrophysiological characteristics of INa in isolated human ventricular myocytes have not yet been fully described. We characterized the human ventricular INa of enzymatically isolated myocytes using whole cell voltage-clamp techniques. Sixty myocytes were isolated from ventricular specimens obtained from 22 patients undergoing open-heart surgery. A low temperature (17 degrees C) and Na+ concentration in the external solution (5 or 10 mM) allowed good voltage control and facilitated the measurement of INa. Cs+ was substituted for K+ in both internal and external solutions to block K+ currents, and F- was added to the internal solution to block Ca2+ current. INa was activated at a voltage threshold of approximately -70 mV, and maximal inward current was obtained at approximately -30 mV (holding potential = -140 mV). The voltage dependence of steady-state INa availability (h infinity) was sigmoidal with half inactivation occurring at -97.3 +/- 1.1 mV and a slope factor of 5.77 +/- 0.10 mV (n = 60). We did not detect any significant differences in these parameters in cells from patients with a variety of disease states, with or without congestive heart failure. The overlap in voltage dependence of h infinity and Na+ conductance suggested the presence of a Na+ "window" current. An inactivation time course was voltage dependent and was fitted best by the sum of two exponentials. The rate of recovery from inactivation also was voltage dependent and fitted by the sum of two exponentials.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238419 TI - Influence of vagotomy on the baroreflex sensitivity in anesthetized dogs with experimental heart failure. AB - In a previous study (Wang, W., J. S. Chen, and I. H. Zucker. Circ. Res. 68: 1294 1301, 1991), we showed a depression in baroreflex gain in dogs with pacing induced heart failure. This depression was not accompanied by a decrease in the central gain (carotid sinus nerve stimulation vs. renal nerve activity). These previous experiments were carried out on animals that were vagotomized and aortic denervated. In the present study these experiments were repeated, and the data were analyzed both in the intact and vagotomized state. Dogs were cardiac paced at 250 beats/min until heart failure was noted. Sham dogs were used as controls. The carotid sinuses were isolated, and the aortic nerves were sectioned. Activity from a renal sympathetic nerve (RSNA) was recorded with arterial pressure and carotid sinus pressure (CSP) during bilateral step increases in CSP from 25 to 300 mmHg. Mean arterial pressure (MAP) and RSNA responses to carotid sinus pressurization and to carotid sinus nerve stimulation were tested before and after bilateral vagotomy, and curves describing these relationships were constructed. Before vagotomy, the peak response relating stimulation frequency to MAP was significantly depressed in dogs with heart failure (-18.3 +/- 4.7 vs. 37.0 +/- 2.2%, P < 0.001). In addition, the CSP-MAP curves were depressed in dogs with heart failure. Frequency-RSNA curves were not altered in heart failure. There were no significant changes in the baroreflex gain responses to pressurization or electrical stimulation after vagotomy in either sham or heart failure groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238420 TI - Endothelin-1 produces pulmonary vasodilation in the intact newborn lamb. AB - The vascular endothelium mediates, in part, pulmonary vascular tone. Because endothelin-1 (ET-1), a paracrine hormone produced by vascular endothelial cells, has vasoactive properties, we investigated the hemodynamic effects of intrapulmonary injections of ET-1 in eight intact newborn lambs at rest and during pulmonary hypertension. At rest, ET-1 (50-1,000 ng/kg) did not change pulmonary arterial pressure. During pulmonary hypertension induced by the infusion of U46619 (a thromboxane A2 mimic), ET-1 (50-1,000 ng/kg) produced a selective dose-dependent decrease in pulmonary arterial pressure (5.8 +/- 3.9 to 32.9 +/- 6.9%; P < 0.05). Similarly, during pulmonary hypertension induced by alveolar hypoxia, ET-1 (50-500 ng/kg) produced a selective dose-dependent decrease in pulmonary arterial pressure (7.2 +/- 3.6 to 26.1 +/- 3.3%; P < 0.05). The decrease in pulmonary arterial pressure produced by ET-1 (250 ng/kg) was attenuated by N omega-nitro-L-arginine (an inhibitor of endothelium-derived nitric oxide synthesis, 23.7 +/- 3.4 vs. 12.5 +/- 4.7%; P < 0.05) and by glibenclamide (an ATP-gated potassium-channel blocker, 25.2 +/- 5.0 vs. 9.6 +/- 5.3%; P < 0.05) but not by meclofenamic acid (an inhibitor of prostaglandin synthesis). ET-1 is a pulmonary vasodilator during pulmonary hypertension in the intact newborn lamb. The vasodilating properties are mediated, in part, by release of endothelium-derived nitric oxide, and by activation of ATP-gated potassium channels. PMID- 8238421 TI - Regulation of glycolytically fueled Ca2+ uptake in smooth muscle plasmalemmal vesicles by phosphorylation. AB - A highly purified plasma membrane vesicular preparation from porcine antrum had an endogenous protein kinase activity with substrates of molecular weights of 11, 15, 20.5, 25, 35, 44, 155, and 230 x 10(3). Phosphorylation of the plasma membranes by the endogenous protein kinase activity resulted in a stimulation of initial rates of Ca2+ uptake into inside-out vesicles, which was associated with an increase in the maximum velocity of the Ca2+ pump with no apparent changes in the half-maximal effective concentration for calcium. Because we have previously reported that a membrane-associated glycolytic system may preferentially provide ATP to fuel the Ca2+ pump (9), we examined the effects of phosphorylation on Ca2+ uptake when glycolysis was the sole source of ATP for the pump. We found that the stimulation of Ca2+ uptake by phosphorylation was more pronounced when Ca2+ uptake was supported by glycolysis rather than 2 mM ATP. When ATP was added at a level similar to that produced by endogenous glycolysis, the stimulation of Ca2+ uptake by phosphorylation was comparable to when glycolysis supported the Ca2+ pump. Our observations suggest that the dynamic range (up to threefold) for regulation of the plasmalemmal Ca2+ pump by phosphorylation is considerably larger than previously reported and thus likely to be of physiological significance. PMID- 8238422 TI - Effects of ischemia on sarcoplasmic reticulum and contractile myofilament activity in human myocardium. AB - The effects of global ischemia on the contractile system and on sarcoplasmic reticulum (SR) function were studied by measuring the isometric tension and the SR Ca2+ release activity of chemically skinned cardiac fiber preparations from seven patients undergoing open-heart surgery. Ten minutes of ischemia caused 1) a decrease in the myofilament sensitivity to Ca2+ (expected Ca2+ concentration giving half-maximal tension; from 0.69 +/- 0.04 to 1.38 +/- 0.06 microM, n = 7) and in the cooperativity index (Hill coefficient; from 2.61 +/- 0.45 to 0.92 +/- 0.15, n = 7), 2) a decrease in myosin light chain phosphorylation, and 3) a 300% increase in the threshold caffeine concentration for SR Ca2+ efflux channel activation, with a 30% reduction in the rate of Ca2+ release by caffeine at threshold concentrations and a 23% reduction in the rate of release by 20 mM caffeine. After preincubation with 5 microM trifluoperazine, a calmodulin antagonist, the caffeine threshold of ischemic and control cardiac muscle became comparable. Most changes were reversed by reperfusion, while the caffeine threshold was still two times greater than control. These results indicate that ischemia caused alterations of the cardiac muscle contractile apparatus and the SR that were reversed only after reperfusion. PMID- 8238423 TI - Effects of cardiac contraction and cavity pressure on myocardial blood flow. AB - Regional impairment of cardiac contraction uncouples force generation from left ventricular pressure (LVP) and may alter the determinants of the phasic pattern and transmural distribution of coronary flow. In anesthetized, open-chest dogs with maximal coronary vasodilation, we studied the effects of abolishing local contraction and changing cavity pressure on phasic myocardial inflow and net transmural flow in a region of left ventricular free wall. With contraction present, the normalized amplitude of distal phasic coronary velocity (NAmp) was not significantly different at normal vs. low LVP (1.00 vs. 0.92 +/- 0.09, respectively, intracoronary lidocaine, however, NAmp varied with LVP (1.62 +/- 0.25 at normal LVP, 0.85 +/- 0.22 at low LVP, P < 0.0001). With contraction present, inner-to-outer flow ratio was not consistently different at normal vs. low LVP (0.47 +/- 0.15 vs. 0.64 +/- 0.28, respectively, P = NS) but was consistently higher at low than at normal LVP with contraction absent (1.01 +/- 0.30 vs. 1.84 +/- 0.38, respectively, P < 0.0001). During uniform global function, contraction is the main determinant of phasic amplitude and transmural distribution of myocardial flow. When regional contraction is abolished, allowing passive deformation of the wall during systole, LVP assumes a powerful role. PMID- 8238424 TI - A comparison of cyclopiazonic acid and ryanodine effects on cardiac muscle relaxation. AB - We investigated cardiac muscle behavior after inhibition of either sarcoplasmic reticulum (SR) Ca2+ release or SR Ca2+ uptake. Mechanics of 35 rat papillary muscles were studied after either ryanodine 10(-7) M (n = 11) or cyclopiazonic acid (CPA) 10(-5) M (n = 14) and compared with a control group containing the solvent alone (n = 10). We measured the maximum extent of shortening (delta L) of the preloaded twitch (delta Lp), and the normalized total force (TF) of the full isometric twitch (TFi). The peak lengthening velocity (Vl) of the preloaded twitch (Vlp) and the normalized negative peak force derivative of the fully isometric twitch (-DFi) tested the lusitropic state. With the influence of shortening and/or load on relaxation taken into account, analysis of relaxation was performed using 1) Vlp-to-delta Lp and magnitude of -DFi-to-TFi ratios and 2) slopes of the Vl-delta L and magnitude of -DF-TF relationships over the entire continuum of load. Ca(2+)-release inhibition with ryanodine induced a negative inotropic effect and a decrease in Vlp from 2.7 +/- 0.2 to 1.4 +/- 0.2 Lmax/S, where Lmax is the initial length at the peak of the length-active tension curve (P < 0.001). The Vlp-to-delta Lp ratio and the slope of the Vl-delta L relationship were preserved, indicating that ryanodine was devoid of intrinsic relaxant effect under isotonic conditions. Ca(2+)-uptake inhibition with CPA had no inotropic effect but decreased Vlp from 2.9 +/- 0.1 to 2.2 +/- 0.1 Lmax/s (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238425 TI - Downregulation of C-receptor by natriuretic peptides via ANP-B receptor in vascular smooth muscle cells. AB - The natriuretic peptide system comprises at least three ligands, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP) and three receptors, the ANP-A receptor, the ANP-B receptor, and the clearance (C) receptor. In the present study, the regulation of natriuretic peptide receptors by their ligands was investigated in cultured rat vascular smooth muscle cells (SMC). The treatment of vascular SMC with ANP, BNP, or CNP decreased the C-receptor density significantly, and the rank order of potency for this downregulation was CNP > ANP > BNP. This rank order was the same as that for guanosine 3',5'-cyclic monophosphate (cGMP) production by ANP, BNP, or CNP in vascular SMC and also the same as that for the ligand selectivity of the ANP-B receptor rather than the C-receptor. The incubation of vascular SMC with 8-bromoguanosine 3',5'-cyclic monophosphate significantly decreased the C receptor density and its mRNA expression. These results suggest that the down regulation of the C-receptor by natriuretic peptides is induced not by the binding of natriuretic peptides to the C-receptor but by the activation of the ANP-B receptor-cGMP pathway. PMID- 8238426 TI - Connections between the pontine reticular formation and rostral ventrolateral medulla. AB - Pontine reticular formation (PRF) neurons provide tonic excitatory drive to sympathetic nerves and are involved in cardiovascular control [K. Hayes and L. C. Weaver. Am. J. Physiol. 263 (Heart Circ. Physiol. 32): H1567-H1575, 1992]. However, connections between the PRF and the well-known vasomotor region in the rostral ventrolateral medulla (RVLM) are unknown. In propofol (Diprivan)- anesthetized rats we investigated arterial pressure, heart rate, and renal nerve responses to microinjection of glycine (1.0 M, 60 nl) into the PRF before and after injection of the synaptic blocking agent cobalt chloride (4.0 mM, 200 nl) into the RVLM. Glycine injections into the PRF caused decreases in arterial pressure, heart rate, and discharge of renal sympathetic nerves. Synaptic blockade of the RVLM almost eliminated cardiovascular and sympathetic responses to glycine injections into the PRF and blocked somatosympathetic reflexes in the renal nerve. Cobalt injections into the RVLM had very small effects on basal renal nerve firing, arterial pressure, or heart rate. These results suggest that the neurons within the RVLM relay influences from the PRF to sympathetic preganglionic neurons. Because injections of the excitatory amino acid antagonist, kynurenate, into the RVLM also interrupted responses to blockade of the PRF and blocked somatosympathetic reflexes, glutamate is a likely neurotransmitter from the PRF to the RVLM and for somatosympathetic reflexes. PMID- 8238427 TI - A direct negative inotropic effect of acetylcholine on rat ventricular myocytes. AB - Acetylcholine (ACh) decreased the contraction of rat ventricular cells within 20 s. ACh (3.1 x 10(-8) M) produced a half-maximal effect and 10(-6) M ACh produced a maximal effect (a 23.8 +/- 5.4% decrease; mean +/- SE, n = 11). During a 3-min exposure to ACh, the inotropic effect faded. Parallel changes were observed in action potential duration: ACh caused an immediate shortening of the action potential, but then the effect faded with time. The changes in action potential duration were the cause of the changes in contraction, because ACh had no effect on contraction when the contractions were triggered by voltage-clamp pulses of constant duration. The changes in action potential duration were the result of the activation of a K+ current (iK,ACh) by ACh. During an exposure to ACh, this current faded as a result of desensitization. iK,ACh was 6.3 times smaller in ventricular than in atrial cells. This may explain why the negative inotropic effect of ACh on atrial cells was greater: 1.0 x 10(-8) M ACh produced a half maximal effect on atrial cells, and 10(-6) M ACh produced a near maximal effect (a 74.5 +/- 9.5% decrease; n = 4). PMID- 8238428 TI - Arteriolar dilation mediated by capsaicin and calcitonin gene-related peptide in rats. AB - In addition to altering vascular tone by stimulating primary afferent nerves and acting through reflex pathways, capsaicin acts locally. We examined effects of topically applied capsaicin on arteriolar diameter in striated muscle and tested the hypothesis that capsaicin can alter microvascular tone by releasing substance P (SP) or calcitonin gene-related peptide (CGRP). In anesthetized rats, the right cremaster muscle was exposed and suspended in a tissue bath filled with a physiological salt solution. Diameters of third-order arterioles were displayed and measured using in vivo video microscopy. In 17 of 20 rats, addition of capsaicin (3 x 10(-7) M) to the bath dilated arterioles (85 +/- 14% above control). Failure of a second administration of capsaicin to produce a sustained dilation in 6 of 7 arterioles that had previously dilated to capsaicin is consistent with the hypothesis that this agent causes depletion of an endogenous vasodilator. Pretreatment with an SP inhibitor did not alter capsaicin-induced dilation. CGRP (1 x 10(-10) to 2 x 10(-8) M) caused dilation similar to that caused by capsaicin. Pretreatment with a CGRP inhibitor to the bath prevented capsaicin-induced dilation, but not constriction. These results suggest that capsaicin can dilate microvessels by releasing CGRP, which can modulate tone. PMID- 8238429 TI - Modulation of calcium homeostasis in cultured rat aortic endothelial cells by intracellular acidification. AB - Acidosis produces vasodilation in a process that may involve the vascular endothelium. Because synthesis and release of endothelium-derived vasodilatory substances are linked to an increase in cytosolic calcium concentration ([Ca2+]i), we examined the effect of intracellular acidification on cultured rat aortic endothelial cells loaded either with the pH-sensitive probe carboxy seminaphthorhodafluor-1 or the Ca(2+)-sensitive fluorescent probe indo 1. The basal cytosolic pH (pHi) of endothelial monolayers in a 5% CO2-HCO3- buffer was 7.27 +/- 0.02 and that in a bicarbonate-free solution was 7.22 +/- 0.03. Acidification was induced either by removal of NH4Cl (delta pHi = -0.10 +/- 0.02), changing from a bicarbonate-free to a 5% CO2-HCO3(-)-buffered solution at constant buffer pH (delta pHi = -0.18 +/- 0.03), or changing from a 5% to a 20% CO2-HCO3- solution (delta pHi = -0.27 +/- 0.07). Regardless of the method used, intracellular acidification increased [Ca2+]i as indexed by indo 1 fluorescence. The increase in [Ca2+]i induced by changing from a 5 to a 20% CO2-HCO3- solution was not significantly altered by removal of buffer Ca2+ either before or after depletion of bradykinin- and thapsigargin-sensitive intracellular Ca2+ stores. Thus intracellular acidification of vascular endothelial cells releases Ca2+ into the cytosol either from pH-sensitive intracellular buffer sites, mitochondria, or from bradykinin- and thapsigargin-insensitive intracellular stores. This Ca2+ mobilization may be linked to endothelial synthesis and release of vasodilatory substances during acidosis. PMID- 8238430 TI - Microvessel PO2 measurements by phosphorescence decay method. AB - A system is described for the in vivo noninvasive measurement of intravascular PO2 at the microscopic level. Under special circumstances the method can also be used to measure interstitial PO2. The PO2 determination is based on the O2 dependent quenching of phosphorescence of palladium-porphyrins bound to albumin. This compound was injected intravenously in the dosage of 30 mg/kg body wt and dissolved in saline to a concentration of 15 mg/ml. The phosphorescence emission was excited by epi-illumination with a strobe xenon arc and measured by a photomultiplier in a well-defined tissue area as small as 15 x 30 microns. A selected portion of the phosphorescence decay was fitted by a single exponential, and the Stern-Volmer equation was used to calculate PO2. Calibration was performed in vitro using saline and blood and was in agreement with previous reports. In vivo observations were made in normal tissue regions from the unanesthetized hamster transparent skin fold chamber preparation. The method allows PO2 determinations, in the range of 0-80 mmHg, in microvessels with diameters of 15-100 microns. Simultaneous transillumination of the tissue also allows measurement of vessel diameter and red blood cell velocity in the same vessels. PMID- 8238431 TI - Effect of local change in O2 saturation of hemoglobin on cerebral vasodilation from hypoxia and hypotension. AB - We tested the effect of certain newly synthesized allosteric modifiers of hemoglobin on the dilation induced by arterial hypoxia, arterial hypotension, and arterial hypercapnia in cerebral arterioles of anesthetized cats equipped with cranial windows for the observation of the cerebral microcirculation. The allosteric modifiers of hemoglobin are isomers of 2-(aryloxy)-2-methylpropionic acid. They shift the oxygen dissociation of hemoglobin to the right, thereby facilitating the local release of oxygen. When these compounds were applied topically by superfusion at a rate of 1 ml/min in a concentration of 0.1 mM, they had no significant effect on baseline arteriolar diameter but reduced significantly the vasodilation from arterial hypoxia and arterial hypotension. They did not influence the vasodilation from arterial hypercapnia. Spectrophotometric measurements of optical densities from pial veins 50-80 microns in diameter indicated that the superfusion with the allosteric compounds reduced hemoglobin oxygen saturation both during room air breathing and during hypoxia. We conclude that the vasodilations from arterial hypoxia and arterial hypotension are mediated by local oxygen-dependent mechanisms. The allosteric modifiers of hemoglobin may be useful as tools in investigating oxygen-dependent mechanisms. PMID- 8238432 TI - An optical device to measure the dynamics of apex rotation of the left ventricle. AB - Systolic counterclockwise rotation of the left ventricular apex with respect to the base has been defined as left ventricular (LV) twist or torsion. If rotation of the base during systole is small, we hypothesized that the dynamics of twist can be well characterized through the measurement of apical rotation alone. A device was designed to measure apical rotation in a simpler, more direct fashion, providing continuous high-fidelity dynamic measurements. The device consists of a light source, a position-sensitive diode, and a small rotating mirror that is coupled to the apex of the heart by a wire. As the wire rotates, apical rotation (measured in degrees) can be calculated from the position of the deflected light beam. The timing of apical rotation was compared with simultaneous recordings of electrocardiogram, LV pressure, and LV diameter measurements. An initial clockwise rotation (untwist) of 4 +/- 2 degrees (SD) occurred during isovolumic contraction followed by counterclockwise rotation (twisting) through ejection, reaching maximum apical rotation of -15 degrees just before the end of systole. Rapid untwisting during isovolumic relaxation was shown with near-complete dissipation of twist by the first one-third of the diastolic filling period. Caval occlusion caused a downward and leftward shift of the pressure-apical rotation loops, and more twist/untwist was seen to occur during the respective isovolumic contraction and relaxation periods. We conclude that this device provides precise timing and definition of rapid changes during isovolumic contraction and relaxation, confirms results obtained by more laborious methods, and provides an easy method to measure the dynamics of apical rotation continuously during interventions such as load changes. PMID- 8238433 TI - Enhanced oxygenation in vivo by allosteric inhibitors of hemoglobin saturation. AB - The in vivo effects on hemoglobin (Hb)-O2 affinity and tissue PO2 were investigated after intraperitoneal administration of 2-[4-(((dichloroanilino) carbonyl)methyl)phenoxyl]-2-methyl propionic acid (RSR4; 150 mg/kg) or its 3,5 dimethyl derivative (RSR13; 300 mg/kg) in C3Hf/Sed mice. The Hb-O2 dissociation curve was plotted from tail vein blood samples using an O2 dissociation analyzer before and up to 160 min after compound administration. Twenty to 40 min after injection, the PO2 at 50% saturation of hemoglobin (Hb P50) increased by a mean of 25% (range 18-31%) after RSR4 and 53% (range 36-76%) after RSR13. Tissue PO2 was continuously measured using an O2 microelectrode in thigh muscle before and up to 40 min after RSR4 or RSR13 injection. Twenty to 40 min after administration, tissue PO2 increased by a mean of 78% (range 30-127%) after RSR4 and 66% (range 39-97%) after RSR13 administration in anesthetized mice. No change in tissue PO2 was seen in anesthetized controls. PMID- 8238434 TI - Inhibition by a stable factor derived from neutrophils of endothelium-dependent relaxation in rat aorta. AB - Polymorphonuclear leukocytes (PMNs) may play an important role in many pathophysiological states. The effect of a factor derived from PMNs on endothelium-dependent relaxation was studied using rat aortic rings in organ chambers. PMNs were obtained from cardiac surgical patients and healthy volunteers. After incubation in Krebs solution for 3 h, supernatants of PMN suspensions were isolated and used to pretreat the aortic rings for 30 min. The results showed that the supernatants derived from 1 x 10(4) to 5 x 10(6) cells/ml PMNs produced a concentration-dependent inhibition of endothelium-dependent relaxation to acetylcholine but not endothelium-independent relaxation to sodium nitroprusside. The effect could not be prevented by oxygen free radical scavenger superoxide dismutase (150 U/ml), catalase (1,200 U/ml), or mannitol (20 mM) used alone or in combination. Heating the supernatants at 95 degrees C for 30 min did not reduce the inhibitory effect. L-Arginine at 3 x 10(-5) to 3 x 10(-3) M did not significantly reverse the inhibitory effect of the PMN-derived factor. In conclusion, this study reveals that a heat-stable factor derived from human PMNs potently inhibits acetylcholine-induced endothelium-dependent relaxation but not sodium nitroprusside-induced endothelium-independent relaxation in rat aorta. This inhibitory effect is not caused by oxygen free radicals, a limitation of nitric oxide precursor or other unstable factors. PMID- 8238435 TI - Modulation of coronary smooth muscle KCa channels by Gs alpha independent of phosphorylation by protein kinase A. AB - The occupancy of beta-receptors in the smooth muscle membrane of the coronary arteries produces vasodilation and a concomitant hyperpolarization. Large conductance calcium-activated K (KCa) channels are likely to be involved in such hyperpolarization, since they are densely distributed in coronary myocytes, and they are targets of beta-adrenergic stimulation in other smooth muscles. We sought to explore if coronary smooth muscle KCa channels are modulated by beta agonists and we studied the mechanisms of their activation. We found that KCa channels reconstituted into lipid bilayers were activated in the presence of GTP by the beta-adrenergic receptor agonist isoproterenol. KCa channels were also stimulated on non-specific activation of an endogenous G protein(s) with guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), on addition of a purified activated stimulatory G protein (Gs alpha), and when the catalytic subunit of protein kinase A (PKA) was added. Inhibition of PKA activity prevented KCa channel stimulation by PKA, but not by endogenous G protein or by exogenous Gs alpha. These results indicate that beta-adrenoceptor activation of coronary smooth muscle KCa channels results from a dual control: 1) a membrane delimited, possibly direct action of Gs, independent of PKA-mediated phosphorylation; and 2) by PKA-dependent phosphorylation. PMID- 8238436 TI - Transient outward current in young and adult diseased human atria. AB - In human atrial fibers, the action potential undergoes a major developmental change in shape in the months immediately after birth. Transient potassium outward currents, which may affect the shape of the action potential, have been studied using a whole cell voltage-clamp technique with single cells from the atria of young patients aged 3-60 mo. Transient outward current (I(to)) amplitude was measured as the difference between the peak current and the steady-state outward current (I(late)) at the end of the voltage step. The density of I(to) was significantly reduced in adult diseased cells (n = 18) compared with normal cells (n = 21) in a large range of potential and absent in young diseased cells (n = 13). Furthermore, the I(late) recorded in young cells was significantly greater (23.7 +/- 5.74 pA/pF at 60 mV) than in adult normal cells (12.71 +/- 2.25 pA/pF at 60 mV), whereas I(late) was not significantly different between both groups of adult cells. Nevertheless, a 4-aminopyridine-sensitive I(to) has been recorded in young cells. A decrease in the frequency of clamp steps (from 0.1 to 0.01 Hz) did not reactivate a I(to) in young cells. This absence of 4 aminopyridine-sensitive Ito in young cells probably results from either a normal developmental change of this current or from pathological alterations like those described in adult diseased atria. PMID- 8238437 TI - The heart works against gravity. AB - The circulatory systems of vertebrate animals are closed, and blood leaves and returns to the heart at the same level. It is often concluded, therefore, that the heart works only against the viscous resistance of the system, not against gravity, even in vascular loops above the heart in which the siphon principle operates. However, we argue that the siphon principle does not assist blood flow in superior vascular loops if any of the descending vasculature is collapsible. If central arterial blood pressure is insufficient to support a blood column between the heart and the head, blood flow ceases because of vascular collapse. Furthermore, the siphon principle does not assist the heart even when a continuous stream of blood is flowing in a superior loop. The potential energy gained by blood as it is pumped to the head is lost to friction in partially collapsed descending vessels and thus is not regained. Application of the Poiseuille equation to flow in collapsible vessels is limited; resistance depends on flow rate in partially collapsed vessels with no transmural pressure difference, but flow rate is independent of resistance. Thus the pressure developed by the heart to establish a given flow rate is independent of the resistance occurring in the partially collapsed vessels. The pressure depends only on the height of the blood column and the resistance in the noncollapsed parts of the system. Simple laboratory models, involving water flow in collapsible tubing, dispel the idea that the siphon principle facilitates blood flow and suggest that previously published results may have been affected by experimental artifact. PMID- 8238438 TI - Physiological responses of freeze-tolerant and -intolerant frogs: clues to evolution of anuran freeze tolerance. AB - Freeze tolerance in the wood frog, Rana sylvatica, is promoted by multiple, integrated physiological responses to ice forming within body tissues. By analyzing the freezing responses of the sympatric, but freeze intolerant, leopard frog (R. pipiens), we sought clues to the evolution of anuran freeze tolerance. Physiological responses critical to R. sylvatica's freeze tolerance, such as the synthesis and distribution of the cryoprotectant glucose, protective dehydration of organs, and deferred cardiac failure, were present, but comparatively less pronounced, in R. pipiens. Both species were innately tolerant of hyperglycemia. Glucose supplements did not enhance the freezing viability of R. pipiens, although in vitro tests of cryoprotectant efficacy revealed that glucose and glycerol provided comparable protection to erythrocytes of both species. We conclude that the evolution of freeze tolerance in R. sylvatica is not only promoted by its desiccation tolerance and the fortuitous biophysical consequences of freezing (e.g., exothermic induction of cardioacceleration and moderation of cooling rate) but also involves a progressive enhancement of fundamental physiological stress responses. PMID- 8238439 TI - Pioglitazone attenuates hypertension and inhibits growth of renal arteriolar smooth muscle in rats. AB - Hypertension is frequently associated with insulin resistance. We evaluated the effects of pioglitazone, an agent that increases insulin sensitivity, on the development of hypertension in the Dahl salt-sensitive (Dahl-S) rat and in the one-kidney, one-clip Sprague-Dawley rat. We also evaluated the effects of pioglitazone on growth of cultured preglomerular renal arteriolar smooth muscle cells. In Dahl-S rats fed a 3% NaCl diet, tail systolic blood pressures and direct arterial pressures were lower (P < 0.05) in pioglitazone-treated (20 mg/kg daily by gavage for 3 wk) than in control rats (n = 10 rats/group). In one kidney, one-clip Sprague-Dawley rats, systolic blood pressures were also lower in pioglitazone-treated animals (P < 0.0001). In vitro, proliferation of arteriolar smooth muscle cells was stimulated (P < 0.01) by insulin, epidermal growth factor (EGF), and fetal calf serum (FCS); pioglitazone (5 microM) reversibly inhibited (P < 0.01) insulin-, EGF-, and FCS-induced proliferation. Pioglitazone (0.01-100 microM) also inhibited insulin- (1 mU/ml), EGF- (100 ng/ml), and 5% FCS-induced [3H]thymidine incorporation in a concentration-dependent manner (P < 0.01). Thus pioglitazone attenuated the development of hypertension in the Dahl-S rat and the one-kidney, one-clip rat. The ability of pioglitazone to inhibit growth of vascular smooth muscle may contribute to its hypotensive effect. PMID- 8238440 TI - Modulation of systemic interleukin-6 induction by central interleukin-1. AB - Centrally administered interleukin (IL)-1 [both alpha and beta forms, 200 ng/rat intracerebroventricularly (icv)] results in a larger increase in serum IL-6 than after systemic injection, indicating the brain's role in the acute phase response. This action was prevented by the IL-1-receptor antagonist IL-1Ra (20 micrograms/rat icv). Neither antiserum against corticotropin-releasing factor (CRF) nor the alpha-helical-CRF antagonist (25 micrograms/rat icv) affected IL-6 induction by central IL-1 beta, which, however, was significantly prevented by the synthetic glucocorticoid dexamethasone [3 mg/kg intraperitoneally (ip)]. Naloxone, the opiate antagonist, but not naloxone methiodide, its quaternary salt that does not penetrate the blood-brain barrier (both administered at 10 mg/kg ip), antagonized this action of IL-1 beta. After intracerebroventricular IL-1 beta, IL-6 levels in brain areas (striatum, hippocampus, hypothalamus) were extremely low, suggesting that the brain does not significantly contribute to IL 6 synthesis in this condition. The results show that induction of high serum IL-6 levels by central IL-1 beta is mediated by brain IL-1 receptors and is sensitive to inhibition by corticosteroids. The inhibitory effect of naloxone suggests that central opiates are required for this action of IL-1 beta. PMID- 8238441 TI - Control of bilateral seminal emissions from ejaculatory ducts by a lumbar splanchnic nerve. AB - To investigate the route of efferent signals for seminal emissions from ejaculatory ducts (SEEDs), canine lumbar splanchnic nerves (LSNs) were electrically stimulated. SEED was confirmed by visual verification of seminal flow into the exposed posterior urethra. In intact dogs, electrical stimulation of an LSN caused bilateral SEEDs in 13 of 16 dogs examined, with a greater volume at the stimulated side. After transection of a unilateral hypogastric nerve, bilateral SEEDs occurred by electrical stimulation of the contralateral LSN in 11 of 14 dogs with a greater volume at the stimulated side and by the stimulation of the ipsilateral LSN in 13 of 15 dogs with a greater volume at the contralateral side. Contraction pressure of the epididymal tail under the same conditions harmonized with the above results. We conclude that each LSN generates bilateral SEEDs by sending signals to bilateral epididymal tails and that some of the signals through each LSN cross to the other side at the caudal mesenteric plexus and/or the prostatic plexus. PMID- 8238442 TI - Development of meadow voles is influenced postnatally by maternal photoperiodic history. AB - The importance of maternal photoperiodic history during lactation on the postnatal development of meadow vole pups was tested by cross-fostering pups on the day of birth between photorefractory females (housed for 26 wk in a 10:14-h light-dark cycle before mating) and photosensitive females (housed for 2 wk in a 10:14-h light-dark cycle before mating). Control pups were cross-fostered to females with the same photoperiodic history as the gestational dam. Somatic growth and food intake of cross-fostered pups were influenced by the photoperiod of the postnatal lactating dam. Pre- and postnatal maternal photoperiodic history interacted with the sex of the offspring to influence reproductive development. Pelage development was determined exclusively by the prenatal maternal photoperiodic history. The impact of a postnatal maternal influence on pup development of photoperiod-sensitive traits is discussed. PMID- 8238443 TI - Plasma epinephrine appearance and clearance rates in fetal and newborn sheep. AB - Newborn plasma catecholamine levels are elevated compared to fetal life. Whether this reflects increased catecholamine secretion after birth or decreased clearance is not known. To determine the plasma appearance and clearance rates for catecholamines during the transition to postnatal life, we compared plasma epinephrine appearance and clearance rates in fetal sheep before birth and in newborns after delivery. Plasma epinephrine appearance and clearance rates were measured by radiotracer analysis in eight fetuses at 127 +/- 1 days of gestation and, after cesarean delivery, at 130 +/- 1 days of gestation. There was no difference in plasma epinephrine appearance rate during the fetal (26 +/- 4 ng.kg 1 x min-1) or newborn studies (31 +/- 5 ng.kg-1 x min-1). The fetal plasma epinephrine clearance rate (131 +/- 13 ml.kg-1 x min-1) was significantly higher than newborn plasma epinephrine clearance rate (40 +/- 3 ml.kg-1 x min-1). Thus fetal plasma epinephrine appearance rate is not different from appearance rate in the immediate newborn period when catecholamine levels are higher than during most other physiological circumstances. The increase in circulating catecholamine levels at birth is due in part to a significant decrease in clearance rate. It is likely that removal of the placental contribution to whole body clearance accounts for much of the difference observed in fetal and newborn clearance rates. PMID- 8238444 TI - Plasma beta-hydroxybutyrate after octanoate challenge: attenuated ketogenic capacity in neonatal swine. AB - Suckling neonatal pigs (NP, 24 h old) do not exhibit elevated blood ketone bodies (KB). Mature swine have relatively high KB under certain conditions, suggesting an ontogeny of ketogenesis. Thus we evaluated the hypothesis that NP possess a relatively attenuated ketogenic capacity vs. weaned pigs (WP) and mature pigs (MP). Fasted animals were given an intraperitoneal dose of octanoate (C8), and plasma beta-hydroxybutyrate (beta-OHB) and C8 were monitored over 180 min. Newborn (NR, 24 h old) and mature rabbits (MR, > 1 yr old) were also compared. Linear regressions of plasma beta-OHB (microM) vs. plasma C8 (microM) were calculated for C8 < 1,000 microM. There was a significant linear relationship of beta-OHB regressed against C8 in all ages of pigs (P < 0.001) and in NR (P = 0.024). The slope for NP (0.08) was one to two orders of magnitude below slopes for older pigs (WP = 1.19 and MP = 0.78, P < 0.01 vs. NP), NR *6.97, P < 0.05), and MR (4.04, NS). The beta-OHB peak in NP (40.9 +/- 4.4 microM) was 1-8% of the maxima in other animals (P < 0.05) despite a C8 maximum (2.3 +/- 0.3 mM) similar to that of WP (1.9 +/- 0.7 mM) and MR (2.9 +/- 1.2 mM) (P > 0.05, NS). The data are consistent with the hypothesis that NP have a poor capacity for ketogenesis. PMID- 8238445 TI - In vivo evidence for adrenal catecholamine release mediated by nonnicotinic mechanism: local medullary effect of VIP. AB - The aim of the present study was 1) to develop a model in anesthetized dogs in which local infusion of a given substance could be made to the adrenal gland without any systemic effects and 2) to show in this model the potential existence of a nonnicotinic mechanism involved in adrenal catecholamine secretion. Plasma catecholamine concentrations were determined by an high-performance liquid chromatography method. The local infusion into the left adrenolumbar artery (0.5 ml/min for 1 min) of either dimethylphenylpiperazinium (0.3-3.0 micrograms/ml) or vasoactive intestinal peptide (0.1-10.0 micrograms/ml) resulted in dose-dependent increases in both epinephrine and norepinephrine secretions. Neither aortic pressure nor plasma catecholamine levels altered during the drug infusion. The net increases in adrenal catecholamine secretion obtained with dimethylphenylpiperazinium were abolished by the nicotinic blockade with pentolinium (2 mg/ml, 0.5 ml/min for 2 min). However, the net catecholamine responses to vasoactive intestinal peptide remained unaffected in the presence of the same dose of pentolinium. The results suggest that there exists a nonnicotinic mechanism that may be implicated in the local regulation of medullary catecholamine secretion in the dog adrenal gland. This model may be a useful tool for studying local physiological role(s) of various neurotransmitters and neuromodulators in the adrenal secretory function in vivo. PMID- 8238446 TI - The effect of pregnancy on the response to the TxA2/PGH2 analogue U-46619 in rabbits. AB - We compared the hemodynamic actions of U-46619, a stable thromboxane A2 (TxA2) prostaglandin H2 (PGH2) analogue, in nonpregnant (NP) rabbits with those observed in late pregnant (P) rabbits. An intravenous injection of U-46619 (10 micrograms) to each of eight NP chronically instrumented rabbits (mean body weight 3.4 kg) induced an immediate (1 min) and reversible fall of cardiac output (CO, 66%) and mean arterial pressure (MAP, 41%, both P < 0.01). P rabbits (n = 6, mean body weight 3.8 kg), however, responded with an elevation of MAP (5%, P < 0.02) upon intravenous injection of the drug (10 micrograms), while CO remained unchanged. The fall of CO in NP rabbits was associated with the temporary disappearance of a fraction of circulating platelets between the superior vena cava and the aortic arch. The number of platelets at 30 and 60 s after U-46619 was reduced (P < 0.05) by 14 and 20% respectively in the aortic blood, whereas caval platelet counts were unchanged until 90 s (-6%, P < 0.05). In contrast, intraaortic administration of this drug (10 micrograms) to NP rabbits resulted in neither thrombocytopenia nor hypotension. U-46619 (10-30 micrograms i.v.) caused no decrease in platelet count in the aorta of P rabbits. In vitro, U-46619-induced aggregation of platelets harvested from P rabbits was also blunted (P < 0.001). This could not be attributed to reduced affinity or number of platelet thromboxane receptors. The data indicate that U-46619 induces a fall of arterial pressure simultaneous with intravascular platelet aggregation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238447 TI - Circadian rhythms of thermoregulation in the squirrel monkey (Saimiri sciureus). AB - Phase or amplitude differences between rhythms in heat production (HP) and heat loss (HL) have been suggested to account for the circadian rhythm in body temperature (Tb). To describe the relationships among these rhythms in a primate, five unrestrained squirrel monkeys (1.0-1.3 kg) were studied using combined direct and indirect calorimetry, with telemetry of Tb and activity, in a 24-h light-dark cycle (LD 12:12) at 25 +/- 0.5 degrees C. Dry (D; sensible) HL, evaporative (E) HL, HP (oxygen consumption and CO2 production), Tb, and activity were measured at 10-min intervals for a week. Tb, activity, HP, and HL displayed daily rhythms, peaking during the light period. Although the timing of peaks was not significantly different, the diurnal increase in Tb was seen to result from a delayed increase in DHL, and possibly, EHL, relative to increased HP. The nocturnal decrease in Tb was due to different time courses of decrease in HP and HL, with no clear lag in HL. The rhythm in Tb therefore resulted from both phase and time course differences in HP and HL rhythms. PMID- 8238448 TI - Determinants of fasting hypertriglyceridemia in ventromedial hypothalamic obesity in rats. AB - The present study investigated the mechanism of fasting hypertriglyceridemia in ventromedial hypothalamic (VMH) obesity by measurement of post-heparin plasma lipoprotein lipase (LPL) activity, triglyceride secretion rate (TGSR), and plasma insulin. One week after VMH lesions, when the lesioned rats were gaining weight rapidly (the dynamic phase), they showed normal plasma triglyceride levels with increased plasma LPL activity and TGSR. There was a positive correlation between hyperinsulinemia and elevated plasma LPL activity or TGSR in VMH-lesioned rats, while no correlation was observed in control rats. Ten weeks after VMH lesions, when the rats had become obese and reached a steady-state weight gain (the static phase), they showed hypertriglyceridemia with increased plasma LPL activity and TGSR. There was, again, a positive correlation between hyperinsulinemia and elevated plasma LPL activity or TGSR in VMH-lesioned rats. These results suggest a possible mechanism of fasting hypertriglyceridemia in these rats; in the dynamic phase adipose tissue adequately takes up circulating triglyceride because the tissue has sufficient take-up capacity and hence hypertriglyceridemia does not develop. In the static phase the tissue cannot adequately take up circulating triglyceride because of a limitation of its capacity, resulting in hypertriglycemia despite enhanced triglyceride secretion and increased LPL activity in both phases. PMID- 8238449 TI - Effects of anesthesia on cardiac and renal sympathetic nerve activities and plasma catecholamines. AB - The effects of pentobarbital and chloralose on cardiac sympathetic nerve activity (CSNA), renal sympathetic nerve activity (RSNA), arterial pressure (AP), and heart rate (HR) were examined using conscious cats. Arterial blood was sampled intermittently to measure plasma epinephrine. Pentobarbital (25-30 mg/kg iv) decreased CSNA, RSNA, AP, and HR. The reduction of CSNA (71 +/- 7%) was larger and lasted longer than that of RSNA (33 +/- 12%). Chloralose (40-50 mg/kg iv) decreased CSNA 66 +/- 9% and HR, increased RSNA 127 +/- 122%, and did not affect AP. The baroreflex relationship between AP and CSNA was examined by increasing AP to 145 mmHg and decreasing AP to 55 mmHg. Both pentobarbital and chloralose shifted the AP-CSNA relationship curve downward and blunted the slope of the curve, indicating that both drugs attenuate tonic and baroreflex cardiac sympathetic outflow. Pentobarbital and chloralose reduced plasma epinephrine, suggesting a decrease in adrenal sympathetic nerve activity. It is concluded that pentobarbital or chloralose affects differentially sympathetic outflows to different organs such as the heart, kidney, and adrenal gland. PMID- 8238450 TI - Decreased potency of contraction to alpha-adrenoceptor stimulation in renal arteries from obese hypertensive dogs. AB - The contractile potencies of the alpha-adrenoceptor agonist phenylephrine (PhE) and two nonadrenergic vasoconstrictors, prostaglandin F2 alpha (PGF2 alpha) and endothelin, were evaluated in renal arterial muscle strips isolated from obese hypertensive (OH) dogs. Responses were compared with those obtained from renal arteries isolated from lean normotensive (LN) dogs. Identical dose-response curves were produced by PGF2 alpha in arteries from OH and LN dogs. This was also true for endothelin. However, the vasoconstrictor potency of PhE for arteries from OH dogs was approximately 2.5-fold less than that for arteries from LN dogs. This difference was not dependent on endothelial integrity. Although they were less potent to PhE, arteries from OH dogs did not produce weaker maximum contractile responses than arteries from LN dogs; responses produced by maximum K+ depolarization (S(o)) were approximately 2 x 10(5) N/m2, and responses to the maximally effective concentrations of PhE, PGF2 alpha, and endothelin were approximately 0.97-, 0.50-, and 0.87-fold S(o), respectively. In addition to the rightward shift in contractile potency to PhE, arteries from OH dogs precontracted with a maximum PhE concentration relaxed more to a high nitroglycerin concentration than did arteries from LN dogs. At a PhE concentration that produced equivalent maximum force responses, arteries from OH dogs had a lower rate of muscle shortening than did arteries from LN dogs, suggesting reduced cross-bridge activation in the arteries from OH dogs. These data suggest that alpha-adrenoceptor-induced activation was selectively downregulated in renal arteries from OH dogs. PMID- 8238451 TI - Time-dependent phrenic nerve responses to carotid afferent activation: intact vs. decerebellate rats. AB - The objectives were to determine 1) respiratory responses to carotid chemoreceptor inputs in anesthetized rats and 2) whether the cerebellar vermis plays a role in these responses. A carotid sinus nerve was stimulated (20 Hz) with five 2-min trains, each separated by approximately 3 min. During stimulation, respiratory frequency (f), peak amplitude of integrated phrenic nerve activity (integral of Phr), and their product (f x integral of Phr) immediately increased. As stimulation continued, integral of Phr progressively increased to a plateau [short-term potentiation (STP)], but f and f x integral of Phr decreased [short-term depression (STD)] to a value still above control. Upon stimulus termination, integral of Phr progressively decreased but remained above control; f and f x integral of Phr transiently decreased below baseline. After the final stimulation, integral of Phr remained above control for at least 30 min [long-term facilitation (LTF)]. Repeated 5-min episodes of isocapnic hypoxia also elicited STP, STD, and LTF. Vermalectomy lowered the CO2-apneic threshold and eliminated LTF. In conclusion, carotid chemoreceptor activation in rats elicits STP and LTF similar to that in cats; the vermis may play a role in LTF. A new response, STD, was observed. PMID- 8238452 TI - Effect of CO2/pH on the aldosterone response to hypoxia in bovine adrenal cells in vitro. AB - Acidosis increases and hypoxia decreases aldosterone production from the adrenal zona glomulerosa in vivo, in situ, and in vitro. These effects appear to be located at different steps in the steroidogenic process. Because respiratory acidosis and hypoxemia are common sequelae of chronic lung disease, the present experiments evaluated the interaction of hypoxia and CO2 (with uncompensated or compensated extracellular pH) on aldosteronogenesis in vitro. Bovine adrenal zona glomerulosa cells were stimulated with angiotensin II (ANG II) or adenosine 3',5' cyclic monophosphate under room air control (21% O2-0% CO2), CO2 per se (21% O2 10% CO2), hypoxia per se (10% O2-0% CO2), and the combination of CO2 and hypoxia (10% O2-10% CO2). Furthermore, under CO2, pH was either allowed to decrease from 7.2 to 6.8 (uncompensated) or its decrease was minimized (> 7.05) with NaOH (compensated). CO2 without pH compensation led to a significant increase in ANG II-stimulated aldosterone release; when the decrease in pH was minimized, CO2 inhibited ANG II-stimulated aldosterone release. Hypoxia inhibited aldosterone release; the inhibitory effect of hypoxia predominated when combined with CO2. In the presence of cyanoketone, pregnenolone production from endogenous precursors (early pathway) was unaffected. However, the conversion of corticosterone to aldosterone (late pathway) was inhibited by low O2 but unaffected by CO2. It is concluded that the inhibitory effect of low O2 on the late pathway predominates over the effects of uncompensated or compensated simulated respiratory acidosis on aldosteronogenesis. PMID- 8238453 TI - Blood pressure effects of iontophoretically applied bioactive hormones in the anterior forebrain of the rat. AB - In the course of electrophysiological investigations using iontophoresis, we observed that in specific regions of the forebrain even these minute applications of peptide and steroid hormones can influence systemic blood pressure. In urethan anesthetized male Wistar rats, with a catheter in the femoral artery, iontophoretic application of the peptide hormones angiotensin II (ANG II), vasopressin (AVP), and atrial natriuretic peptide (ANP) and the steroid hormone aldosterone (ALDO) was effective at locations in the midline septum, the triangularis nucleus of the septum, and the subfornical organ (SFO). Increases in blood pressure (of up to 15 mmHg) were observed after ANG II and AVP, decreases after ALDO, and either an increase or a decrease, depending on the location, after ANP. There was no clear evidence of an antagonistic effect of ANP on ANG II induced neuronal or blood pressure responses. In addition to demonstrating the potency of these hormones even when they are restricted to tiny volumes of tissue, the present results demonstrate that the medial ventral region of the anterior forebrain may be included in the same baroreceptive circuit as the SFO (and organ vasculosum of the lamina terminalis) and hence be involved in the regulation of blood volume and perhaps in the sensing of and corrective responses to extracellular thirst. PMID- 8238454 TI - Effect of a central CRF antagonist on cardiovascular and thermoregulatory responses induced by stress or IL-1 beta. AB - We investigated the role of central corticotropin-releasing factor (CRF) in the development of cardiovascular and thermal responses induced by stress or by interleukin-1 beta (IL-1 beta) in free-moving rats. Intracerebroventricular (icv) injection of alpha-helical CRF9-41 (10 micrograms), a CRF receptor antagonist, significantly attenuated hypertension, tachycardia, and a rise in body temperature induced by cage-switch stress, a mild stress. However, icv injection of alpha-helical CRF9-41 (10 micrograms) had no effect on hypertension, tachycardia, or fever induced by intraperitoneal (ip) injection of IL-1 beta (2 micrograms/kg) or icv prostaglandin E2 (PGE2, 100 ng). In contrast, icv injection of alpha-helical CRF9-41 (10 micrograms) significantly attenuated hypertension, tachycardia, or fever induced by icv injection of IL-1 beta (20 ng). The present results suggest that central CRF has an important role in the development of the cage-switch stress-induced responses, but it does not seem to contribute to the hypertension, tachycardia, and fever induced by ip IL-1 beta or by central PGE2. However, it is possible that when IL-1 beta directly acts on the central nervous system, some of its actions are mediated by central CRF. PMID- 8238455 TI - Hypothalamic peptide regulation of ACTH secretion from sheep pituitary. AB - The relative abilities of the hypothalamic peptides corticotropin-releasing factor (CRF), arginine vasopressin (AVP), oxytocin (OT), and angiotensin II (ANG II) to stimulate adrenocorticotropic hormone (ACTH) secretion from cultured sheep anterior pituitary cells were studied. Incubation of cells with CRF, AVP, and OT, but not ANG II, was associated with increased ACTH secretion. CRF and AVP were equally effective in stimulating ACTH release at 0.1 nM, but larger doses of each resulted in distinctly different ACTH secretory patterns. The minimally effective dose of OT was 10 nM; greater doses of this peptide resulted in ACTH secretory responses similar to those measured after addition of AVP. Cotreatment with ANG II did not affect the ACTH-secretory response to CRF, AVP, or OT. These data confirm that AVP is a potent stimulus for ACTH secretion from sheep anterior pituitary in vitro and also show that CRF is effective in low concentrations in releasing ACTH. In contrast, the data do not support a regulatory role for ANG II in stimulating ACTH release directly from sheep corticotroph cells. PMID- 8238456 TI - Conservation of photoperiod-responsive mechanisms in humans. AB - In animals, circadian pacemakers respond to seasonal changes in day length by making corresponding adjustments in the durations of diurnal and nocturnal periods of circadian rhythms; these adjustments mediate effects of photoperiod on breeding and other seasonally recurring phenomena. Little is known about photoperiod responses of human circadian pacemakers. To investigate this question, we recorded and compared circadian rhythm profiles of 15 individuals after chronic exposures to short (8 h) and long (14 h) nights. As occurs in animals, durations of nocturnal periods of active melatonin secretion (11.9 +/- 1.6 vs. 10.3 +/- 1.3 h, df = 14, t = 4.583, P < 0.0005, paired t test), high prolactin secretion (12.9 +/- 2.1 vs. 9.9 +/- 2.2 h, df = 11, t = 2.917, P < 0.01), and sleep (10.6 +/- 0.8 vs. 7.6 +/- 0.4 h, df = 14, t = 17.122, P < 0.0005) were longer after exposure to long nights than after short ones. Durations of nocturnal periods of low rectal temperature (11.6 +/- 2.3 vs. 9.5 +/ 1.6 h, df = 12, t = 3.912, P < 0.001) and rising cortisol secretion (10.8 +/- 1.6 vs. 9.3 +/- 1.9 h, df = 14, t = 3.130, P < 0.005) were also longer. Some of these differences persisted during 24-h periods of enforced wakefulness in constant dim light, indicating that prior exposure to the two regimes induced abiding changes in the timing of internal processes, such as circadian pacemaker oscillations, that control the durations of nocturnal and diurnal periods of the rhythms. PMID- 8238457 TI - Fetal ACTH and blood pressure responses to thromboxane mimetic U-46619. AB - This study was performed to test the hypothesis that thromboxane A2 stimulates increases in fetal adrenocorticotropic hormone (ACTH), vasopressin, or renin secretion and affects fetal cardiovascular function by an action on the fetal central nervous system. We infused a stable synthetic analogue of thromboxane A2, U-46619, into one common carotid artery or inferior vena cava or infused saline into one common carotid artery in chronically catheterized fetal sheep between 127 and 140 days gestation. We found that intracarotid but not intravenous infusions of U-46619 at a rate of 750 ng/min stimulated increases in fetal plasma ACTH concentration. Infusions of U-46619 at both sites increased fetal blood pressure; the infusion into the carotid arterial blood produced a more rapid increase in blood pressure and a significant decrease in central venous pressure. None of the infusions altered plasma vasopressin concentration or plasma renin activity, blood gases, hematocrit, or plasma cortisol concentration. We conclude that thromboxane A2 stimulates fetal ACTH, but not vasopressin or renin, secretion via an action within the area perfused by carotid arterial blood. Thromboxane A2 increases blood pressure via an action at the fetal central nervous system, as well as via a direct vasoconstrictor action in the systemic circulation. PMID- 8238458 TI - Augmented glucoregulatory hormone concentrations during exhausting exercise in mildly iron-deficient rats. AB - We hypothesized that augmented responses of glucoregulatory hormones in iron deficiency would enhance liver and muscle glycogenolysis, leading to increased gluconeogenic precursor (lactate) supply and upregulation of hepatic gluconeogenesis. Female weanling rats were randomly placed on either a mildly iron-deficient (-Fe; 15 mg Fe/kg diet) or an iron-sufficient (+Fe; 50 mg Fe/kg diet) diet for 4 wk and studied at rest and during exhaustive treadmill running. Hemoglobin was 9.0 +/- 0.2 and 13.1 +/- 0.3 g/dl in -Fe and +Fe, respectively, after 3.5 wk of dietary iron deficiency. Arterial plasma epinephrine (Epi), norepinephrine (NE), adrenocorticotropic hormone (ACTH), corticosterone, insulin, and glucagon levels were similar at rest in both groups, as were liver, gastrocnemius, and superficial and deep vastus medialis glycogen levels. Liver and kidney phosphoenolpyruvate carboxykinase (PEPCK) activities were similar in both groups. Maximum O2 consumption was decreased (22%) in -Fe. Respiratory exchange ratio (CO2 production/O2 consumption) was unaffected at rest but increased at maximum O2 consumption in -Fe. Time to exhaustion during a standardized running test (13.4 m/min, 0% grade) was decreased 45% in -Fe (63 +/- 5 vs. 116 +/- 10 min). During exercise, euglycemia was maintained in both groups, but blood lactate was elevated in -Fe. The mean net glycogen utilization during exercise was increased in liver (43%), soleus (33%), and superficial vastus medialis (106%) and decreased in the gastrocnemius (36%) in -Fe. Liver and kidney PEPCK activities were increased similarly at exhaustion in both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238459 TI - Gastric loads and cholecystokinin synergistically stimulate rat gastric vagal afferents. AB - Both gastric preloads and exogenous cholecystokinin (CCK) administration inhibit food intake, and combinations of preloads and CCK suppress feeding to a greater degree than either stimulus delivered alone. A role for the vagus nerve in mediating CCK's inhibition of food intake has been proposed, and gastric vagal afferent fibers respond to both gastric loads and local CCK infusions. To examine whether combined load and CCK stimuli may synergistically augment gastric neural afferent activity at the level of the peripheral vagus, we have examined the gastric vagal afferent responses (n = 8) to a range of gastric saline loads (1, 2, and 3 ml) and exogenous close celiac arterial CCK (10 and 100 pmol) when administered alone or in combination. Gastric loads ineffective in eliciting a significant increase in vagal afferent activity when administered alone became effective when combined with doses of CCK that were subthreshold for the production of a vagal afferent response. Gastric loads that alone were effective in producing a significant vagal afferent response yielded an even greater response when administered in combination with both subthreshold and suprathreshold doses of CCK. These data demonstrate that, in rats, signals produced by combined gastric load and exogenous CCK administration are integrated peripherally and interact synergistically. These results suggest that signals arising from the vagus may provide sufficient information for the synergistic inhibition of food intake produced by combinations of gastric loads and exogenous CCK. PMID- 8238460 TI - Interleukin-8 modulates feeding by direct action in the central nervous system. AB - Interleukin-8 (IL-8) is released in response to infection, inflammation, and trauma. The most important stimuli for IL-8 release during these pathological processes are IL-1, tumor necrosis factor, and bacterial lipopolysaccharide (endotoxin), factors that have been shown to suppress feeding. In the present study, the participation of IL-8 on the central regulation of feeding was investigated. Intracerebroventricular (icv) microinfusion of recombinant human IL 8 (rhIL-8, 1.0-100 ng/rat) suppressed the short-term (2-h) food intake. The most effective dose of rhIL-8, 20 ng, decreased 2-h food intake by 25% and nighttime food intake by 23%. Intracerebroventricular microinfusion of anti-rhIL-8 antibody (200 and 500 ng) blocked the effect of 20 ng rhIL-8 on 2-h and nighttime food intakes. Computerized analysis of behavioral patterns for the 2-h period demonstrated a specific reduction of meal size (by 33%), whereas meal frequency and meal duration were not affected after the icv microinfusion of 20 ng rhIL-8. This short-term food intake suppression by icv rhIL-8 was accompanied by a small, but significant, increase in cerebrospinal fluid-brain and rectal temperatures. Intraperitoneal administration of rhIL-8 in doses equivalent to those administered centrally had no effect on food intake. The results suggest that IL 8 acts directly in the central nervous system to decrease feeding. This effect of IL-8 may contribute to the food intake suppression frequently accompanying pathological processes. PMID- 8238461 TI - Unaltered cerebral blood flow during hypoglycemic activation of the sympathochromaffin system in humans. AB - To determine if increases in plasma epinephrine and norepinephrine caused by hypoglycemia are associated with increments in cerebral blood flow (CBF), we measured CBF with positron emission tomography in normal humans at ambient fasting arterial plasma glucose levels and at clamped plasma glucose levels of 5.3, 3.5, and 2.8 mmol/l using the hyperinsulinemic, stepped hypoglycemic clamp technique. Despite significant increases in plasma epinephrine to 7,340 +/- 350 (SE) pmol/l and in plasma norepinephrine to 3.32 +/- 0.35 nmol/l, mean hemispheric CBF (41 +/- 1, 49 +/- 1, 48 +/- 2, and 51 +/- 3 ml x 100 g-1 x min-1) remained constant at the ambient (5.2), 5.3, 3.5, and 2.8 mmol/l glycemic levels, respectively. Furthermore, there was no correlation between CBF and either plasma epinephrine levels ranging from 160 to 10,580 pmol/l or plasma norepinephrine levels ranging from 0.56 to 5.10 nmol/l. Failure to demonstrate any dose-response relationship between plasma catecholamine levels and CBF argues against their primary role in cerebrovascular control during hypoglycemia. PMID- 8238462 TI - Postpyloric stimuli are necessary for the normal control of meal size in real feeding and sham feeding rats. AB - When 0.8 M sucrose is sham fed, intake increases progressively on consecutive tests and then plateaus. Interspersing two real feeding tests between sham feeding tests prevents this increase in sham intake [J. D. Davis and G. P. Smith. Am. J. Physiol. 259 (Regulatory Integrative Comp. Physiol. 28): R1228-R1235, 1990]. This is consistent with an acquired control of ingestion based on an association between an oropharyngeal conditioned stimulus (CS) and a postingestional unconditioned stimulus (UCS). To determine if this postingestional UCS has a gastric or postgastric origin (or a combined one), we confined ingested 0.8 M sucrose to the stomach on real feeding and sham feeding tests by closing the pylorus with an inflatable Silastic cuff. Rats were tested on seven sham feeding tests separated by two real feeding tests and then on six consecutive sham feeding tests. In contrast to our previous results [Davis and Smith. Am. J. Physiol. 259 (Regulatory Integrative Comp. Physiol. 28): R1228 R1235, 1990], intake increased progressively on the real feeding and both types of sham feeding tests. We conclude that both gastric and postgastric stimuli act as a UCS for the acquired control of meal size. PMID- 8238463 TI - Renal response to atrial stretch during pregnancy in conscious rats. AB - Chronic indwelling balloons were placed at the junction of the right atrium and the right superior vena cava of virgin female rats. The renal response to discrete standardized atrial stretch was tested in the conscious animals. The rats were then mated, and the renal response was retested at days 7, 14, and 20 of pregnancy. On day 21, the rats were killed, and the pressure-volume relationship of the right atria was measured. In response to atrial stretch, there was an increase in urine volume, urine sodium output, and urine potassium output and a decrease in free water clearance in the virgin rats. Subsequent to mating, these responses were completely abolished at all stages of pregnancy. The renal responses to atrial stretch were also abolished during pseudopregnancy. Central venous pressure did not change when the intracardiac balloons were inflated; there were also no changes in resting central venous pressure during pregnancy. There were no significant changes in unstressed atrial volume or in atrial compliance during pregnancy. PMID- 8238464 TI - An IL-1 receptor and an IL-1 receptor antagonist attenuate muramyl dipeptide- and IL-1-induced sleep and fever. AB - It is hypothesized that the somnogenic and pyrogenic effects of muramyl dipeptide (MDP) are mediated via enhanced interleukin-1 (IL-1) production. To test this hypothesis the effects of intracerebroventricular (icv) administration of a recombinant human soluble type I IL-1 receptor (sIL-1r) and of the IL-1 receptor antagonist (IL-1ra) on MDP-induced sleep and fever were evaluated in rabbits. The sIL-1r recognized rabbit IL-1 beta, but it did not affect sleep or brain temperature across the dose range tested (1-50 micrograms) when injected icv into normal rabbits. Pretreatment of rabbits with 50 micrograms sIL-1r or 10 micrograms IL-1ra blocked human recombinant IL-1-enhanced nonrapid eye movement (NREM) sleep and fever. Thus both the sIL-1r and the IL-1ra were effective antagonists of IL-1 actions. When the animals were pretreated with either 50 micrograms sIL-1r or with 10 or 100 micrograms of the IL-1ra, the somnogenic effects of 150 pmol MDP were attenuated. However, the sIL-1r had little effect on MDP-induced febrile responses. These results suggest that the sIL-1r and the IL 1ra can function as antagonists of IL-1 actions in vivo and that MDP-induced sleep and fever are partially mediated by IL-1. PMID- 8238465 TI - Modulation of in situ canine intrinsic cardiac neuronal activity by locally applied adenosine, ATP, or analogues. AB - To determine whether adenosine or ATP can modify mammalian intrinsic cardiac neurons, these substances, as well as their analogues 5'-(N-ethylcarboxamido) adenosine (NECA), N6-cyclopentyladenosine (CPA), and beta,gamma-methylene ATP (beta,gamma-mATP), were applied in microliter quantities adjacent to spontaneously active canine atrial ganglionated plexus neurons in 14 anesthetized open-chest dogs. Adenosine, NECA, and CPA induced neuronal responses, neuronal activity being either increased or decreased in 81, 86, and 86% of the sites tested, respectively. Cardiovascular responses were elicited by these agents in 21-31% of neurally active loci. ATP and beta,gamma-mATP elicited neuronal responses in 100 and 70% of tested loci, respectively. Associated cardiovascular responses were elicited by ATP and beta,gamma-mATP in 35 and 18% of the sites, respectively. After acute decentralization of the intrinsic cardiac nervous system, neuronal responses were elicited by purines in 73% of the previously active sites, while cardiovascular responses were either attenuated or eliminated. It is concluded that exogenous purine nucleosides and nucleotides can modulate the activity generated by in situ intrinsic cardiac neurons presumably by acting on P1 and P2 purinoreceptors. Furthermore, these data indicate that purine sensitive intrinsic cardiac neurons are involved in cardiac regulation. PMID- 8238466 TI - Alterations in brain hexokinase activity associated with heart failure in rats. AB - This study examined the activity of discrete regions of the brain as assessed with histological localization and photodensitometric quantification of the metabolic enzyme hexokinase in a group of rats with coronary occlusion (HF) and in sham-operated control rats. Three weeks after surgery, the mean left ventricular end diastolic pressure and right atrial pressure were elevated, and left ventricular peak systolic pressure was decreased in the HF group compared with the sham group; these findings are also observed during heart failure. In addition, histological data indicated that there was a 37.6 +/- 2.8% outer and 40.8 +/- 3.1% inner infarct of the myocardium in the group of rats with HF (n = 6). Rats in the control group had no observable damage to the myocardium (n = 6). Accompanying these symptoms of heart failure were significant increases in hexokinase activity in the parvocellular (pPVN, 16.3%) and magnocellular (mPVN, 17.6%) divisions of the paraventricular nucleus of the hypothalamus, and in the locus ceruleus (LC, 17.1%). No changes in hexokinase activity were observed in the median preoptic area, supraoptic nucleus (SON), subfornical organ, or posterior hypothalamus. These results reinforce the idea that heart failure (with coronary occlusion) is associated with changes in specific areas in the brain and that metabolic alterations in the pPVN, mPVN, and LC are likely related to alterations in vasopressin production, blood volume regulation, and sympathoexcitation observed in the heart failure state. PMID- 8238467 TI - Reduced ionic conductance in turtle brain. AB - Whole cell recording techniques were employed to measure whole cell (Gw) and specific membrane (Gm) conductance in turtle and rat pyramidal neurons in slices. Results indicate that rat neurons are 4.2 times more conductive compared with turtle neurons at 25 degrees C, which is accentuated by temperature, so that rat neurons at 37 degrees C are 22 times more conductive than turtle neurons at 15 degrees C. A conductance Q10 of 1.9 was measured for both turtle (15-25 degrees C) and rat (25-35 degrees C) pyramidal neurons. Conductance measurements of turtle pyramidal neurons over 6-9 h of anoxia indicate no statistical difference between Gm or Gw from normoxic control measures. These results indirectly support the concept of low ATP-dependent ion pump activity in the turtle brain as one mechanism for reduced energy expenditure in the normoxic state. PMID- 8238468 TI - Arginine vasopressin produces renal vasodilation via V2 receptors in conscious dogs. AB - In conscious dogs, 36-h water deprivation induced a significant increase in renal blood flow (RBF) with elevation of the plasma arginine vasopressin (AVP) concentration to 9.6 +/- 1.8 pg/ml. To simulate such a condition, a mild elevation of plasma AVP was produced by infusing AVP intravenously at a dose of 0.1 ng.kg-1.min-1 for 20 min. The plasma AVP concentration then increased to 6.8 +/- 0.7 pg/ml. This dose of AVP increased the RBF by 21.7 +/- 2.6% and decreased the renal vascular resistance by 18.1 +/- 2.3% without significant changes in mean arterial pressure, cardiac output, or heart rate. The mechanism of this renal vasodilatory action was examined using newly developed, orally effective, selective AVP antagonists OPC-21268 (a V1-receptor antagonist) and OPC-31260 (a V2-receptor antagonist). In 36-h water-deprived dogs, V2-receptor blockade with OPC-31260 significantly decreased the RBF by 20.5 +/- 2.6% without significant changes in cardiac output. The exogenous AVP-induced renal vasodilatory response tended to be augmented when V1 receptors were blocked by pretreatment with OPC 21268, but the change did not achieve statistical significance. On the other hand, V2-receptor blockade by either pretreatment with OPC-31260 or simultaneous infusion of OPC-31260 inhibited this vasodilatory response. Furthermore, intravenous infusion of 1-desamino-8-D-arginine vasopressin (DDAVP) at a dose of 0.3 ng.kg-1 x min-1 for 20 min significantly increased the RBF by 36.5 +/- 1.7%, and this DDAVP-induced renal vasodilation was inhibited by simultaneous infusion of V2-receptor antagonist.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238469 TI - Impairment of renal function precedes establishment of hypertension in spontaneously hypertensive rats. AB - To determine renal function throughout development of hypertension in the spontaneously hypertensive rat (SHR) and its normotensive counterpart, the Wistar Kyoto rat (WKY), renal clearance studies were performed at 2-wk intervals from 4 to 12 wk and again at 16 wk of age in conscious chronically instrumented rats after recovery (4-6 days) from surgery. The data indicate that the critical period for the development of hypertension in SHR was between 4 and 6 wk of age. Mean arterial pressure sharply increased from 107 +/- 5 (n = 6) to 145 +/- 6 mmHg (n = 6) between 4 and 6 wk of age, did not change between 6 and 10 wk of age, and gradually rose between 10 and 16 wk of age to 183 +/- 3 mmHg. In WKYs, blood pressure increased only slightly from 97 +/- 3 mmHg at 4 wk of age (n = 8) to 110 +/- 3 mmHg at 8 wk of age (n = 8), where it remained through adulthood. Glomerular filtration rate (GFR) in the 4-wk-old SHR was significantly decreased compared with WKY (0.94 +/- 0.03 vs. 1.11 +/- 0.04 ml.min-1 x g wet kidney wt-1), and it recovered to normal level and stabilized by 6 wk of age (1.14 +/- 0.04 ml.min-1 x g wet kidney wt-1). Renal blood flow was lower in the SHR only at 4 and 16 wk; it increased with age in both groups. Renal vascular resistance was higher in the SHR at 4 wk and remained elevated throughout the observation period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238470 TI - Torpor shortens the period of Siberian hamster circadian rhythms. AB - We investigated the influence of ambient and body temperature (Ta and Tb) on circadian rhythms of gonadectomized male Siberian hamsters. Animals that entered torpor (Tb < 30 degrees C) had significantly shorter circadian periods (tau s) than did nontorpid hamsters at a Ta of 13 degrees C (24.17 +/- 0.05 vs. 24.33 +/- 0.04 h). The tau s of homeothermic hamsters were not affected by Ta change. Short term decreases in Tb, rather than changes in Ta, appear to affect tau. Access to activity wheels inhibited expression of torpor in short daylengths and was associated with significant increases in body mass. Running wheel activity can mask or block specific short-day responses. PMID- 8238471 TI - Genetic development of an inbred rat strain with increased resistance adaptation to a hot environment. AB - The purpose of this study was to demonstrate resistance adaptation based on genotypic adaptation and to develop an inbred rat strain with genotypic resistance adaptation to a hot environment. Survival time (ST) at an ambient temperature (Ta) of 42.5 degrees C was determined without appreciable thermal damage. Rats with the longest ST were sibmated over 30 generations and designated FOK. The ST, evaporative water loss, and body water economy increased rapidly during the first 10-15 generations, followed by a more gradual increase. The FOK rat resisted a Ta of 42.5 degrees C for > 5 h; this ability was hereditary. Body size progressively decreased during the early generations. The ST was longer (P < 0.01) and evaporative water loss was significantly higher (P < 0.01) in the FOK rat compared with three other strains. Tail lengths in the FOK rat were slightly shorter than those of three controls (P < 0.01). The heat loss system in the FOK rat may depend on the ability to mobilize and evaporate body fluids efficiently. FOK rats can be used for phenotypic comparisons with other strains, as well as in molecular genetic studies on thermoregulation, osmoregulation, and resistance adaptation to heat using recombinant inbred and recombinant congenic lines. PMID- 8238472 TI - In defense of cell volume? AB - Most cells react vigorously to perturbations in their fluid volume. Although the responses themselves have been well described, little is known about how the volume stimulus is detected and transduced. The relevance of volume-activated phenomena in vitro to volume homeostasis in vivo is not clear. It can be debated whether cells are truly responding to changes in their volume. Perhaps they are reacting to changes in the concentration of a cytoplasmic constituent. This review calls attention to recent literature bearing on these subjects and concludes that, in most cases, cells are not defending their volume but their concentration of cytosolic macromolecules. PMID- 8238473 TI - Effect of osmotic swelling and shrinkage on Na(+)-K+ pump activity in mammalian cardiac myocytes. AB - The effect on the sarcolemmal Na(+)-K+ pump of exposure to anisosmolar solutions was examined using whole cell patch clamping and ion-selective microelectrodes. Na(+)-K+ pump currents were measured in single ventricular myocytes by using pipette Na+ concentrations ([Na]pip) of 0-70 mM. The relationship between [Na]pip and pump current was well described by the Hill equation. The [Na]pip for half maximal pump current (K0.5) was 21.4 mM in isosmolar (310 mosM) solution. K0.5 was 12.8 mM during cell swelling in hyposmolar solution (240 mosM) and 39.0 mM during cell shrinkage in hyperosmolar solution (464 mosM). The maximal pump currents, derived from the best fit of the Hill equation, and the Hill coefficients were similar in isosmolar, hyposmolar, and hyperosmolar solutions. A sustained (> 20 min) decrease in the intracellular Na+ activity developed during exposure of intact papillary muscles to hyposmolar solutions, and a sustained increase developed during exposure to hyperosmolar solutions. We conclude that osmotic myocyte swelling stimulates the sarcolemmal Na(+)-K+ pump at near physiological levels of intracellular Na+, whereas shrinkage inhibits the pump. These changes are due to increases and decreases, respectively, in the apparent affinity of the pump for Na+. PMID- 8238474 TI - Effect of morphine on mesangial immunoglobulin G aggregate kinetics. AB - Because mesangial expansion is considered a precursor of focal glomerulosclerosis, we studied whether morphine can cause mesangial expansion. We used radiolabeled human immunoglobulin G aggregates (125I-ahIgG) to study mesangial kinetics in control and experimental (morphine-treated) rats. Control and experimental rats were administered 125I-ahIgG by tail vein. Serum levels of 125I-ahIgG and uptake of 125I-ahIgG by liver, spleen, and mesangium were determined at 4, 8, 12, 24, and 36 h after 125I-ahIgG administration. Mesangial 125I-ahIgG levels were higher (P < 0.05) at 4 h and at later periods in morphine treated vs. control rats. Naloxone, an opioid antagonist, did not attenuate the morphine-induced mesangial accumulation of 125I-ahIgG. The mean uptake of IgG aggregates was lower in the liver and spleen of morphine-treated rats at 36 h (P < 0.05). In both in vivo and in vitro experiments, ultrastructural studies showed accumulation of IgG-coated gold particles in vesicles, endosomes, and lysosomes. Morphine may have increased the accumulation of 125I-ahIgG in the glomeruli either by increasing the delivery of macromolecules into the mesangium or by altering the exit of macromolecules from the mesangium. PMID- 8238475 TI - Possible specific activation of RNA synthesis in PC-12 cell isolated nuclei by small acidic peptides. AB - Three synthetic peptides, pyro-Glu-Ala-Gly-Glu-Ser-Glu-Asp (Pep A), pyro-Glu-Ala Gly-Glu-Glu-Glu-Ser-Asn (Pep B), and pyro-Glu-Asp-Asp-Ser-Asp-Glu-Glu-Asn (Pep C), bear sequences possibly belonging to components of a naturally occurring family of strongly related small acidic chromatin peptides involved in regulation of gene expression. In a crude nuclear fraction and in purified nuclei from PC-12 cells, Pep A and Pep B activate RNA synthesis, specifically acting on the RNA polymerase II transcription system. On the other hand, Pep C shows an inhibitory effect on RNA synthesis in purified nuclei but an activation in the crude nuclear fraction. Control experiments show that the serum thymic factor does not affect RNA synthesis in the crude nuclear fraction or in purified nuclei. A possible regulation by peptide phosphorylation via casein kinase II (more active in purified nuclei than in the crude nuclear fraction) is discussed. PMID- 8238476 TI - Influence of cloned voltage-gated K+ channel expression on alanine transport, Rb+ uptake, and cell volume. AB - Voltage-gated K+ channels are involved in regulation of action potential duration and in setting the resting membrane potential in nerve and muscle. To determine the effects of voltage-gated K+ channel expression on processes not associated with electrically excitable cells, we studied cell volume, membrane potential, Na(+)-K(+)-ATPase activity, and alanine transport after the stable expression of the Kv1.4 and Kv1.5 human K+ channels in Ltk- mouse fibroblasts (L-cells). The fast-activating noninactivating Kv1.5 channel, but not the rapidly inactivating Kv1.4 channel, prevented dexamethasone-induced increases in intracellular volume and inhibited Na(+)-K(+)-ATPase activity by 25%, as measured by 86Rb+ uptake. Alanine transport, measured separately by systems A and ASC, was lower in Kv1.5 expressing cells, indicating that the expression of this channel modified the Na(+)-dependent amino acid transport of both systems. Expression of the Kv1.4 channel did not alter alanine transport relative to wild-type or sham-transfected cells. The changes specific to Kv1.5 expression may be related to the resting membrane potential induced by this channel (-30 mV) in contrast to that measured in wild-type sham-transfected, or Kv1.4-transfected cells (-2 to 0 mV). Blocking of the Kv1.5 channel by 60 microM quinidine negated the effects of Kv1.5 expression on intracellular volume, Na(+)-K(+)-ATPase, and Na(+)-dependent alanine transport. These results indicate that delayed rectifier channels such as Kv1.5 can play a key role in the control of cell membrane potential, cell volume, Na(+)-K(+)-ATPase activity, and electrogenic alanine transport across the plasma membrane of electrically unexcitable cells. PMID- 8238477 TI - Ca2+ influx via T-type channels modulates PDGF-induced replication of mouse fibroblasts. AB - The role of low-threshold voltage-gated calcium channels (VGCC) in modulating extracellular calcium influx and proliferation was investigated in platelet derived growth factor (PDGF)-stimulated C3H/10T1/2 mouse fibroblasts. Previous studies demonstrated that cell cycle progression after PDGF stimulation was dependent on extracellular calcium influx producing a sustained increase in the intracellular calcium concentration. In this study, PDGF-induced calcium influx, the sustained intracellular calcium increase, and progression to S phase were inhibited by nordihydroguariaretic acid (NDGA), an inhibitor of calcium influx through VGCC. With the use of the whole cell patch-clamp technique to measure calcium currents, NDGA inhibited inward calcium current through low-threshold VGCC, the only VGCC expressed in C3H/10T1/2 fibroblasts. The inhibitory effects of NDGA on calcium influx and cell proliferation each had a mean inhibitory dose of 2-3 microM. Although NDGA also effectively inhibits cyclooxygenase and lipoxygenase, the addition of prostaglandins or leukotrienes could not reverse this inhibition nor could it be replicated by other antioxidants. These data support the hypothesis that low-threshold VGCC can mediate extracellular calcium influx on the stimulation of cell proliferation by PDGF. PMID- 8238478 TI - Voltage-dependent kinetics of Na-Ca exchange current in Ca(2+)-loaded guinea pig heart cells. AB - Voltage-dependent properties of Na-Ca exchange current were revealed with the patch-clamp technique in Ca(2+)-overloaded guinea pig ventricular myocytes in the whole cell configuration. With the assumption that the transient inward current (Iti) is mediated by the Na-Ca exchanger, oscillations of internal Ca2+ concentration ([Ca2+]i) were used to investigate voltage-dependent kinetics of exchange current differences at two [Ca2+]i values. After Iti was elicited by clamping from -45 mV to basic pulses of +10 mV, pairs of equipotential short test pulses were applied during the basic pulse at both the phase of low [Ca2+]i (between two neighboring Iti values) and the phase of high [Ca2+]i (at the peak of Iti). The test pulses were short enough to leave the time course of Iti during the basic pulse approximately unchanged, which allowed study of the voltage dependence of the respective current differences without disturbing the underlying oscillation of [Ca2+]i. The current differences were inward at all potentials between -140 and +70 mV, started from an equal initial value, and obeyed characteristic voltage-dependent time courses: hyperpolarization to potentials negative to -70 mV caused an initial current increase, which was followed by a decay to very small amplitudes or zero with a decay time constant decreasing toward hyperpolarization e-fold per 45.6 mV. Depolarizing pulses caused a decay of the current differences to smaller levels. Respective current differences formed during a slowly decaying current component, following the Ca current spike, showed equal voltage-dependent properties. This indicates that the slowly decaying current component is preferentially also carried by the Na-Ca exchanger.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238479 TI - Microelectrode measurements of pericellular PO2 in erythropoietin-producing human hepatoma cell cultures. AB - On the basis of Fick's law of gas diffusion, it has been proposed that cells in conventional monolayer cultures may be severely hypoxic. Because knowledge of the cellular O2 availability is important for the interpretation of biochemical and toxicological cell culture work, microelectrode measurements of the pericellular PO2 were carried out using the erythropoietin (Epo)-producing human hepatoma cell lines Hep G2 and Hep 3B as an in vitro model. In confluent hepatoma cultures grown in polystyrene dishes and incubated in air with 5% CO2, the pericellular steady-state PO2 was < 1 mmHg. The rates of the production of immunoreactive Epo and lactate were high due to a misproportion between O2 supply and O2 requirements. Epo production decreased when shaken instead of static cultures were studied, or when the O2 concentration in the gas atmosphere was increased gradually up to 95%. In cultures grown on gas-permeable supports, pericellular and gas PO2 values were very similar, with increased Epo production at lowered PO2. In agreement with mathematical models, our experimental data make PO2 measurements desirable for studies of O2-dependent biological functions in cell cultures. PMID- 8238480 TI - Hormone-induced rise in cytosolic Ca2+ in axolotl hepatocytes: extracellular origin and control by cAMP. AB - In amphibian liver, signal transduction of [Arg8]vasotocin (AVT), a "classical" Ca(2+)-dependent hormone in rat liver, is mediated via the generation of adenosine 3',5'-cyclic monophosphate (cAMP) and not via inositol 1,4,5 trisphosphate [Ins(1,4,5)P3]. In isolated hepatocytes from axolotl, hormones that stimulated cAMP formation (the order of efficacy was glucagon > isoprenaline > epinephrine > or = AVT) also provoked a pronounced increase in cytosolic Ca2+, as indicated from changes in fura 2 fluorescence. 8-Bromoadenosine 3',5'-cyclic monophosphate at 100 microM was as potent as maximally effective concentrations of glucagon. Ins(1,4,5)P3 mobilized Ca2+ from the endoplasmic reticulum of saponin-permeabilized axolotl hepatocytes with a half-maximal effect at 0.65 microM, as did GTP (20 microM), even in the absence of polyethylene glycol. However, the hormonally induced increase in cytosolic Ca2+ was not due to a mobilization of the cation from internal stores by Ins(1,4,5)P3, but to an increased inflow from the extracellular medium. We conclude that in axolotl liver, in contrast to rat liver, hormones stimulate the production of cAMP that, in addition to stimulating processes such as glycogenolysis, also regulates the opening of an ion gate in the plasma membrane, which allows the inflow of Ca2+. To our knowledge this is the first demonstration of a second messenger-operated Ca2+ channel in a splanchnic tissue. PMID- 8238481 TI - Immediate cell signal by bone-related peptides in human osteoclast-like cells. AB - We tested whether recognition of bone-related peptides regulates intracellular Ca2+ concentration ([Ca2+]i) of giant cell tumor of bone (GCT). [Ca2+]i was measured in single cells by fura 2 fluorometry. GCT cells were sensitive to bone sialoprotein-II (BSP-II), osteopontin (OPN), and related fragments. Responses consisted of a prompt increase of [Ca2+]i, mostly transient, with a peak followed by a rapid return toward baseline. Responses were not mimicked by bovine plasma fibronectin. Sensitivity of GCT cells to bone peptides was specific, since BALB/3T3 fibroblasts and U-937 histiocytic lymphoma cells with monocytic phenotype failed to respond to BSP-II and OPN fragments. GRGDSP synthetic esapeptide, carrying the Arg-Gly-Asp adhesive motif, and GRGESP (Asp replaced by Glu), but not the GRADSP (Gly replaced by Ala), were active in inducing [Ca2+]i transients as well. Responses were observed also in cells treated with the BSP-II 1C fragment, lacking any known adhesive sequence, indicating that the active peptides inducing [Ca2+]i increments may be multiple. Sensitivity to extracellular matrix peptides was present in a variable fraction of the cells and was downregulated on long-term culture. The mechanism inducing [Ca2+]i elevations was mostly related to Ca2+ release from thapsigargin-sensitive intracellular pools. PMID- 8238482 TI - Cell-specific regulation of cytosolic aspartate aminotransferase by glucocorticoids in the rat kidney. AB - The basal expression and hormonal regulation of cytosolic aspartate aminotransferase (cAspAT) were investigated in the rat kidney. In adrenalectomized animals, the basal activity was highest in the renal cortex and in the inner stripe of the outer medulla (0.1-0.15 U/mg protein). The glucocorticoid analogue dexamethasone increased cAspAT activity about twofold in the cortex and in the inner stripe of the outer medulla but not in the papilla. A half-maximal increase in the activity was achieved at doses of approximately 5 micrograms/100 g body wt. The mineralocorticoid aldosterone did not modify the cAspAT activity. The cell specificity of the hormonal regulation was analyzed by in situ hybridization. In untreated adrenalectomized rats, a cAspAT cRNA probe labeled mainly the inner stripe of the outer medulla. After dexamethasone or hydrocortisone treatment, labeling was uniformly increased in this part of the medulla and was heterogeneously increased in the renal cortex. The specific increase in labeling within the cortex was shown to be confined to the distal convoluted tubule and the thick ascending limb. We conclude that, in addition to widespread basal expression, cAspAT is regulated by glucocorticoids in a highly cell-specific manner in the renal cortex. The enzyme may thus participate in the increased energy metabolism elicited by these hormones in these cells. PMID- 8238483 TI - Inhibition of myosin light chain phosphatase during Ca(2+)-independent vasocontraction. AB - Phorbol 12,13-dibutyrate (PDB) induced a sustained contraction of rat thoracic aorta strip in Ca(2+)-free buffer without significant change in intracellular free Ca2+ concentration. NKH477, a water-soluble forskolin derivative, markedly relaxed the PDB-induced contraction. The PDB-induced contraction was associated with the phosphorylation of 20-kDa myosin light chain (MLC). Two-dimensional phosphopeptide mapping of 20-kDa MLC revealed that approximately 90% of the phosphopeptides was derived from an MLC kinase-catalyzed reaction and approximately 10% was due to phosphorylation by protein kinase C. NKH477 inhibited the PDB-induced phosphorylation of 20-kDa MLC. MLC phosphatase activity of intact aorta strips was inhibited by the treatment with PDB, and the inhibition was recovered by the application of NKH477. These results suggest that the regulation of MLC phosphatase in vascular smooth muscle may play important roles in the PDB-induced contraction and the NKH477-induced relaxation in Ca(2+) free buffer. PMID- 8238484 TI - Time-dependent increase in Ca2+ influx in rabbit abdominal aorta: role of Na-Ca exchange. AB - Exposure of the rabbit abdominal aorta to the combination of high K+ and norepinephrine resulted in a time-dependent increase in the rate of 45Ca influx and 45Ca and 22Na content over that observed after stimulation with either K+ or norepinephrine alone. The increase in 45Ca influx, but not the increase in 22Na content, was extracellular Ca2+ (Cao2+) dependent. This time-dependent increase in 45Ca influx was prevented by incubating the tissue in Na(+)-free medium. Nifedipine inhibited both the initial depolarization-induced 45Ca influx and time dependent increase in 45Ca influx and 22Na content. The effect of nifedipine on time-dependent fluxes was prevented by ouabain. Phorbol dibutyrate mimicked the effects of norepinephrine on 22Na retention and 45Ca fluxes. The effects of phorbol dibutyrate and norepinephrine were not additive. It is concluded that, in rabbit abdominal aorta, norepinephrine plus K+ causes 22Na retention (possibly through inhibition of the sodium pump) and a Cao(2+)- and intracellular Na+ (Nai+)-dependent increase in 45Ca influx. This latter effect is possibly the result of increased Nai(+)-Cao2+ exchange. PMID- 8238485 TI - pH heterogeneity during exercise in localized spectra from single human muscles. AB - We investigated whether pH heterogeneity in skeletal muscle during exercise, observed with 31P nuclear magnetic resonance, represents muscle fiber type heterogeneity. Localized spectra were simultaneously acquired from the soleus, medial, and lateral gastrocnemius using a multivolume localization technique, the Hadamard spectroscopic imaging (HSI) technique. Contamination of nonselected regions to the localized volumes was < 5%. HSI-localized spectra were obtained from the calf muscles of untrained subjects and a small group of athletes. Two plantar flexion exercise protocols were implemented: a "maximal" high frequency protocol and a "steady-state" protocol at low contraction frequency (0.25 Hz). pH heterogeneity was observed in localized spectra of single muscles during both exercise protocols, as indicated by the large Pi line width. During maximal exercise the Pi line width was up to three times wider than the phosphocreatine line width, covering an entire pH unit. During the steady-state exercise, in three subjects the Pi peak clearly resolved into two distinct peaks, one at low pH and one at high pH. As pH heterogeneity was observed in localized spectra of single muscles during both exercise protocols, it most likely reflects the metabolic heterogeneity between fiber types. PMID- 8238486 TI - Characterization and confocal imaging of calponin in gastrointestinal smooth muscle. AB - Calponin isolated from chicken gizzard smooth muscle binds in vitro to actin in a Ca(2+)-independent manner and thereby inhibits the actin-activated Mg(2+) adenosinetriphosphatase of smooth muscle myosin. This inhibition is relieved when calponin is phosphorylated by protein kinase C or Ca2+/calmodulin-dependent protein kinase II, suggesting that calponin is involved in thin filament associated regulation of smooth muscle contraction. To further examine this possibility, calponin was isolated from toad stomach smooth muscle, characterized biochemically, and localized in intact isolated cells. Toad stomach calponin had the same basic biochemical properties as calponin from other sources. Confocal immunofluorescence microscopy revealed that calponin in intact smooth muscle cells was localized to long filamentous structures that were colabeled by antibodies to actin or tropomyosin. Preservation of the basic biochemical properties of calponin from species to species suggests that these properties are relevant for its in vivo function. Its colocalization with actin and tropomyosin indicates that calponin is associated with the thin filament in intact smooth muscle cells. PMID- 8238487 TI - Neutrophil-activating intercrine secreted by porcine platelets is active without proteolytic processing. AB - A new member of the cytokine intercrine alpha-subfamily, porcine neutrophil activating peptide 2 (pNAP-2), was isolated to homogeneity. Amino acid sequencing analysis showed two species of pNAP-2, a long form (pNAP-2-L) and a short form (pNAP-2-S). pNAP-2-L had seven more amino acids at the NH2-terminus than pNAP-2 S. The remaining amino acid sequences of the two molecules were identical. pNAP-2 S shared 65% homology with human neutrophil-activating peptide 2 (hNAP-2) including four cysteines in identical positions. Moreover, the NH2-terminal sequence Glu-Leu-Arg (E-L-R) was conserved in both molecules. Both pNAP-2-L and pNAP-2-S induced mobilization of cytosolic calcium in neutrophils and caused release of granulocyte elastase in a dose-dependent manner, although pNAP-2-L was less active. A desensitization study suggested that both hNAP-2 and pNAP-2-S may act on the same receptor. Whereas human platelets release inactive precursors that can be converted to hNAP-2 by cathepsin G from activated neutrophils, porcine platelets, upon stimulation with thrombin, appear to secrete active forms of pNAP-2. The activated neutrophils are not involved in the generation of pNAP 2. PMID- 8238488 TI - Secretagogue-induced RVD in HSY cells is due to K+ channels activated by Ca2+ and protein kinase C. AB - To maintain cell volume, absorptive epithelia must coordinate ion fluxes associated with transcellular transport with those required for volume regulation. K+ channels are thought to play a central regulatory role in this process. Electronic cell sizing was used to study the regulatory volume decrease (RVD) response of the human salivary ductal cell line HSY. Following a hypotonic challenge, RVD was markedly enhanced by the muscarinic agonist carbachol (half maximal effect approximately 1.4 microM). Carbachol-induced RVD was mimicked by the K+ ionophore valinomycin and inhibited by K+ channel blockers, indicating that it is due to the activation of K+ channels. Carbachol-induced RVD was blocked by maneuvers that blunted the carbachol-induced rise in intracellular Ca2+ concentration ([Ca2+]i) and mimicked by ionomycin-induced increases in [Ca2+]i, but concentrations of ionomycin (100 nM) yielding [Ca2+]i well above carbachol-induced levels were required. However, when treatment with 2.5 nM ionomycin, which produced [Ca2+]i in the carbachol-generated range, was combined with treatment with low concentrations of an active phorbol ester, an enhancement of RVD similar to that observed with carbachol alone was observed. These data provide strong evidence that carbachol-induced RVD in the HSY cells involves K+ channels that are tightly regulated by both [Ca2+]i and protein kinase C. PMID- 8238489 TI - Determination of volume and water permeability of plated cells from measurements of light scattering. AB - Measurements of cell membrane water osmotic permeabilities can be inaccurate because of the technical difficulties inherent to cell volume measurements and because of the presence of an unstirred water layer in contact with the cells. We detail here a method we have developed to quantify transient changes in cell volumes from the intensity of light scattered by cells. For this, we theorize how an unstirred layer originates in a perfusing chamber, and we calculate values for both cell membrane water osmotic permeability and unstirred layer thickness from time transient changes in scattered light. We apply a computer algorithm that finds the best correspondence between experimental data and estimated values. This is done by solving a differential equation governing cell volume changes by numerical integration (Runge-Kutta) and iterating the procedure varying the test values of osmotic permeability and unstirred layer thickness until the best fit is achieved. We exemplify this procedure with experimental results obtained in adherent cultured cells. PMID- 8238490 TI - A computer-based servo system for controlling isotonic contractions of muscle. AB - We have developed a computer-based servo system for controlling isotonic releases in muscle. This system is a composite of commercially available devices: an IBM personal computer, an analog-to-digital (A/D) board, an Akers AE801 force transducer, and a Cambridge Technology motor. The servo loop controlling the force clamp is generated by computer via the A/D board, using a program written in QuickBASIC 4.5. Results are shown that illustrate the ability of the system to clamp the force generated by either skinned cardiac trabeculae or single rabbit psoas fibers down to the resolution of the force transducer within 4 ms. This rate is independent of the level of activation of the tissue and the size of the load imposed during the release. The key to the effectiveness of the system consists of two algorithms that are described in detail. The first is used to calculate the error signal to hold force to the desired level. The second algorithm is used to calculate the appropriate gain of the servo for a particular fiber and the size of the desired load to be imposed. The results show that the described computer-based method for controlling isotonic releases in muscle represents a good compromise between simplicity and performance and is an alternative to the custom-built digital/analog servo devices currently being used in studies of muscle mechanics. PMID- 8238491 TI - Nanomolar ouabain augments caffeine-evoked contractions in rat arteries. AB - Chronic parenteral administration of ouabain to normal rats raises plasma ouabain concentrations to low nanomolar levels and induces hypertension [C. M. Yuan, P. Manunta, J. M. Hamlyn, S. W. Chen, E. Bohen, J. Yeun, F. J. Haddy, and M. B. Pamnani. Hypertension 22: 178-187, 1993 and see also M. P. Blaustein. Am. J. Physiol. 264 (Cell Physiol. 33): C1367-C1387, 1993]. To determine whether rat arteries are sensitive to these low ouabain levels, we tested the effects of various ouabain concentrations on caffeine-evoked contractions (CEC) in rat aortic and small mesenteric artery rings. CEC amplitude was used as a measure of the sarcoplasmic reticulum (SR) Ca2+ content. Ouabain increased CEC in aortic as well as mesenteric artery rings, but the effects in the aorta were difficult to quantitate because the CEC were often oscillatory. Mesenteric artery, under control conditions and after sensitization with 10-30 nM phenylephrine (PE), exhibited biphasic ouabain dose-CEC response curves. Low concentrations of ouabain (0.1-10 nM) caused small significant increases in CEC, but a further effect was observed only with > or = 10 microM ouabain. PE shifted the ouabain dose-response curve toward lower ouabain concentrations; conversely, ouabain shifted the PE dose-response curve toward lower PE concentrations. It appears that nanomolar concentrations of ouabain can influence vascular responsiveness to vasoconstrictors. We conclude that rat vascular smooth muscle contains both high- and low-affinity ouabain receptors, possibly corresponding to Na+ pumps with alpha 3- and alpha 1-subunit isoforms, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238492 TI - Interaction of PDGF and natriuretic peptides on mesangial cell proliferation and endothelin secretion. AB - The present study examined the possible interaction of platelet-derived growth factor (PDGF) and atrial or brain natriuretic peptides (ANP and BNP, respectively) on cellular proliferation and secretion of endothelin-1 in cultured rat mesangial cells. PDGF increased cellular proliferation and endothelin-1 secretion. The protein kinase C (PKC) inhibitors, 1-(5-isoquinolinylsulfonyl)-2 methylpiperazine, staurosporine, and PKC inhibitor peptide, inhibited such stimulation. Rat ANP-(1-28) and rat BNP-45 exhibited clearly dose-related inhibition of PDGF-stimulated cellular proliferation and endothelin-1 secretion. This inhibition by ANP and BNP was paralleled by an increase in the cellular level of guanosine 3',5'-cyclic monophosphate (cGMP). Results indicate that PDGF stimulates cellular proliferation and endothelin-1 secretion in cultured rat mesangial cells by a mechanism probably involving activation of PKC and that ANP and BNP inhibit such stimulation through a cGMP-dependent process. PMID- 8238493 TI - Effects of insulin on the counterregulatory response to equivalent hypoglycemia in normal females. AB - The aim of this study was to determine if insulin could augment the counterregulatory response to equivalent hypoglycemia in normal females similarly to males. Experiments were carried out in nine normal lean overnight-fasted female subjects. Insulin was infused in two separate randomized protocols so that steady-state levels of 794 +/- 62 (low) and 3,620 +/- 476 pM (high) were obtained. Despite an identical plasma glucose level (2.8 +/- 0.1 mM), epinephrine (5.7 +/- 0.9 vs. 3.9 +/- 0.6 nM), norepinephrine (2.7 +/- 0.4 vs. 1.8 +/- 0.3 nM), cortisol (918 +/- 55 vs. 826 nM), and growth hormone (35.8 +/- 3.7 vs. 28.4 +/- 2.7 micrograms/l) were increased (P < 0.05) during high compared with low insulin infusion, respectively. Glucagon and pancreatic polypeptide levels increased significantly but were not different during the two insulin infusions. Hepatic glucose production was increased during the high-compared with low-dose infusions (9.5 +/- 1.1 vs. 5.1 +/- 2.2 mumol.kg-1 x min-1; P < 0.05). Lipolysis, as indicated by the blood glycerol level, increased significantly during high- compared with low-dose insulin infusions (121 +/- 29 vs. 65 +/- 13 microM; P < 0.05). The hormonal and metabolic responses to hypoglycemia were significantly different in females compared with previous results in males.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238494 TI - Systemic and uterine responses to chronic infusion of estradiol-17 beta. AB - Human and ovine pregnancies are associated with increases in plasma levels of estrogens and angiotensin II (ANG II), cardiac output (CO), blood volume (BV), and uterine blood flow (UBF), as well as attenuated ANG II pressor responses. We hypothesized that, in nonpregnant animals, prolonged estradiol-17 beta (E2 beta) treatment would reproduce these endocrine and hemodynamic alterations. Nonpregnant ovariectomized ewes (n = 5) received 5 microgram E2 beta/kg iv followed by 220 micrograms/day for 14 days. Plasma E2 beta increased from 36 +/- 6 to 269 +/- 79 (SE) pg/ml (P < 0.05) during E2 beta treatment, returning to control values 4 days posttreatment. By 3 days of E2 beta, mean arterial pressure (MAP) and systemic vascular resistance (SVR) fell 9 +/- 1 and 29 +/- 1%, whereas heart rate (HR) and CO increased 20 +/- 5 and 26 +/- 1% (P < 0.05). Stroke volume (SV), BV, and plasma volume were unchanged until 7 days of E2 beta, with values rising 17 +/- 5, 13 +/- 3, and 14 +/- 4, respectively (P < 0.05). Although MAP remained similarly depressed (-11 +/- 1%) during week 2 of E2 beta, SVR decreased further (-37 +/- 3%) and was associated with additional increases (P < 0.05) in CO to 44 +/- 5%, reflecting rises in SV (21 +/- 2%) but not HR. Increases in BV correlated with rises in CO (r = 0.55) and SV (r = 0.64) but not HR (r = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238495 TI - Ontogenetic regulation of mouse Ren-2d renin gene in transgenic hypertensive rats, TGR(mREN2)27. AB - TGR(mREN2)27 is a new monogenetic rat model with fulminant hypertension, low kidney renin, and high extrarenal renin gene expression. This study characterizes and compares expression of the Ren-2 gene in TGR(mREN2)27 with that in DBA/2 mice and with renin gene expression in rats. Except in the submandibular gland, the tissue-specific expression of Ren-2 is similar in TGR(mREN2)27 and DBA/2. This demonstrates maintenance of tissue specificity. Organs that are involved in cardiovascular regulation, such as the adrenal gland, kidney, and brain, express the Ren-2 gene before hypertension has developed, consistent with the possibility of a causal relationship between transgene expression in these tissues and hypertension. Because these tissues express the renin gene in nontransgenic rats as well, we suggest that this model can be used to study the regulation of renin gene expression and its role in hypertension at these sites. In addition, as an indication that interactions may exist between blood pressure and renin gene expression, we describe reciprocal changes in blood pressure and Ren-2 mRNA levels in the kidney and brain. PMID- 8238496 TI - Effect of endurance training on plasma free fatty acid turnover and oxidation during exercise. AB - Plasma free fatty acid (FFA) levels tend to be lower and the plasma lipolytic hormone response to prolonged exercise of the same intensity is blunted after endurance exercise training. To determine whether training elicits a corresponding decrease in plasma FFA turnover and metabolism during prolonged exercise, we measured plasma [1-13C]palmitate kinetics and oxidation and respiratory gas exchange in 13 subjects during the latter portion of a 90- to 120 min bout of cycle ergometer work performed before and after 12 wk of alternate day cycling and running. Training increased total fat oxidation during prolonged exercise by 41% (P < 0.005). However, for the final 30-60 min of the cycle ergometer protocol, the rate of 13CO2 production from [1-13C]palmitate oxidation was 27% lower (P < 0.05), the rate of palmitate turnover was 33% less (P < 0.05), and plasma FFA and glycerol concentrations were 32 and 20% lower (P < 0.05), respectively, than in the untrained state. Thus endurance exercise training results in decreased plasma FFA turnover and oxidation during a 90- to 120-min bout of submaximal exercise because of a slower rate of FFA release from adipose tissue. PMID- 8238497 TI - Acute hyperglycemia enhances proteolysis in normal man. AB - The influence of hyperinsulinemic-hyperglycemia on protein and carbohydrate homeostasis was assessed using L-[1-13C]-leucine and [3-3H]glucose combined with open-circuit indirect calorimetry. After a 30-min basal period, healthy human volunteers were subjected to two sequential experimental periods (150 min each) during which insulin was continuously infused at a rate to elicit maximal effects (10.0 mU.kg-1 x min-1, resulting in 220-fold basal levels) in conjunction with an infusion of L-amino acids to maintain euleucinemia. Plasma glucose was maintained near basal (94 +/- 2 mg/dl) during period I and at twofold basal (191 +/- 4 mg/dl) during period II. The endogenous rate of leucine appearance (index of proteolysis in mumol.kg-1 x h-1) dropped by 80% from basal during period I (P < 0.01) but only by 44% during period II. Although hyperglycemia stimulated the rate of proteolysis threefold (18.0 +/- 10.2 to 49.8 +/- 7.2, P < 0.001), neither leucine oxidation nor nonoxidative leucine rate of disappearance (an estimate of protein synthesis) was altered. During both glycemic conditions, the rates of hepatic glucose production were completely suppressed, and this was accompanied by 3- and 11-fold increases in the estimated rates of protein and carbohydrate oxidation, respectively, and an 80% decrease in the rate of lipid oxidation. The estimated rate of glucose storage was increased fourfold during period I and by an additional 2.3-fold (to approximately 10-fold basal) in period II. The present study demonstrates that, during hyperinsulinemia, acute elevations of plasma glucose to two times basal levels result in a marked stimulation of whole body proteolysis during hyperinsulinemia. PMID- 8238498 TI - Glutamine carbon disposal and net glutamine uptake in fetuses of fed and fasted ewes. AB - We have traced ovine fetal glutamine carbon uptake and disposal in 7 chronically catheterized fetuses of fed ewes and 10 fetuses of 48-h fasted ewes. Net fetal glutamine uptake (Fick principle, antipyrine blood flow) was 10.0 +/- 2.0 mumol.kg-1 x min-1 in fed fetuses and 6.4 +/- 1.4 mumol.kg-1 x min-1 in fasted fetuses [not significant (NS)]. However, net fetal glutamine uptake was linearly related to the umbilical vein glutamine level (P < 0.05) in fed and fasted fetuses. In contrast, fetal glutamate transfer to the placenta was 4.0 +/- 0.8 mumol.kg-1.min-1 in the fed state and 2.7 +/- 0.1 mumol.kg-1 x min-1 in the fasted state. Net fetal glutamine uptake and fetal glutamate transfer to the placenta were directly correlated (P < 0.05). Fetal glutamine carbon disposal was measured using a primed continuous infusion of [U-14C]-glutamine over a 3-h period and blood sampling during the last hour of infusion (steady state). Disposal was 20.9 +/- 2.6 mumol.kg-1 x min-1 in the fed state and 18.6 +/- 2.3 mumol.kg-1 x min-1 in the maternal fasted state (NS). Glutamine carbon disposal did not correlate with fetal arterial glutamine levels and was not influenced by maternal nutritional state. PMID- 8238499 TI - Determination of specific activity of labeled ligands by nonlinear regression analysis. AB - The specific radioactivity (SA) of 125I-lysine vasopressin (LVP) was determined by analyzing the binding B (cpm/tube) of variable amounts of tracer T (cpm/tube) to a constant amount of an LVP antibody, in the presence of known quantities L (mol/tube) of LVP standards. The parameters of the equations B = f(F) and B/F = g(T), describing B as a function of free F (cpm/tube) tracer or the ratios B/F as a function of T, were first calculated by nonlinear regression analysis of the results obtained with tracer alone. Then the dependent variables B or B/F were measured in the presence of LVP and analyzed with the same equations by substituting the independent variables F or T with (F + alpha FL) and (T + alpha L), respectively, where alpha (cpm/mol) represents a measure of the SA and FL (FL = L.F/T), free LVP, respectively. The SA was thus treated as an unknown parameter to be calculated by nonlinear regression. This method was compared with the traditional interpolation of the SA from the self-displacement and standard curves. Tracer and ligand were found to have the same affinity for the binding sites, since the set of equations B = f(F + alpha FL) and B/F = g(T + alpha L), describing the binding of the tracer in the presence of LVP and equations B = f(F) and B/F = g(T) to which these equations reduce in the absence of LVP (L = 0), had identical binding parameters. To be valid, any method based on self displacement requires that the tracer and standards have the same affinity for the binding sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238500 TI - Regulation of phosphatidylinositol 3-kinase by insulin in rat skeletal muscle. AB - The presence of phosphatidylinositol 3-kinase (PI 3-kinase) in mammalian skeletal muscle and its response to insulin stimulation were investigated. PI kinase, immunoprecipitated from rat soleus muscle with antibodies directed toward its 85 kDa subunit phosphorylated PI, phosphatidylinositol 4-phosphate [PI(4)P], and phosphatidylinositol 4,5,-bisphosphate [PI(4,5)P2] to yield phosphatidylinositol 3-phosphate [PI(3)P], phosphatidylinositol 3,4,-bisphosphate, and phosphatidylinositol trisphosphate in vitro. PI 3-kinase activity was also immunoprecipitated with antiphosphotyrosine [alpha-Tyr(P)] antibodies and with antibodies raised against IRS-1, a substrate of the insulin receptor protein tyrosine kinase that associates with and activates PI 3-kinase. Incubation of the soleus with insulin in vitro, or injection of insulin into rats in vivo, produced three- to fivefold increases in alpha-Tyr(P)- and alpha-IRS-1-immunoprecipitable PI 3-kinase activity. In nonstimulated soleus muscle, PI 3-kinase activity immunoprecipitated with alpha-IRS-1 or with alpha-Tyr(P) antibodies was evenly distributed between particulate (200,000-g pellet) and soluble fractions. Insulin treatment increased immunoprecipitable PI 5-kinase activity in both fractions, but the increase in alpha-Tyr-(P)-precipitable activity was greater in the particulate fraction, whereas the increase in alpha-IRS-1-precipitable activity was greater in the soluble fraction. In intact soleus muscles incubated with 32PO4, insulin increased the labeling of PI(3)P but did not affect the labeling of PI(4)P or PI(4,5)P2. Activation of PI 3-kinase by insulin was unaffected by prior denervation of the muscle, a manipulation that has been shown to cause both insulin resistance and hypersensitivity in muscles, depending on the parameter measured.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238501 TI - Liver and peripheral tissue glycogen metabolism in obese mice: effect of a mixed meal. AB - Glycogen metabolism in the liver, skeletal muscle, cardiac muscle, and white adipose tissue was studied in gold thioglucose (GTG) obese mice after fasting and during refeeding. Prolonged (48 h) fasted control and GTG mice were refed with standard laboratory diet for 24 h. During fasting and refeeding, the changes in glycogen content and the activity of glycogen synthase I and R and phosphorylase alpha in the liver were similar in lean and GTG mice. However, the glycogen storage in the livers from GTG mice was always greater than that in lean animals. In GTG mice the activity of liver glycogen synthase I and R was significantly higher than that in lean animals 3 and 6 h after refeeding. The activity of liver phosphorylase alpha in GTG mice was higher than that in lean mice after refeeding. There were no significant differences in the glycogen content of white adipose tissue, cardiac muscle, and skeletal muscle from lean and GTG mice during the entire study. The results of this study suggest that increased glycogen storage in the liver is a major alteration in nonoxidative glucose metabolism and contributes to the development of insulin resistance and glucose intolerance in GTG obese mice. PMID- 8238502 TI - Pyruvate dehydrogenase activity and acetyl group accumulation during exercise after different diets. AB - Pyruvate dehydrogenase activity (PDHa) and acetyl group accumulation were examined in human skeletal muscle at rest and during exercise after different diets. Five males cycled at 75% of maximal O2 uptake (VO2 max) to exhaustion after consuming a low-carbohydrate diet (LCD) for 3 days and again 1-2 wk later for the same duration after consuming a high-carbohydrate diet (HCD) for 3 days. Resting PDHa was lower after a LCD (0.20 +/- 0.04 vs. 0.69 +/- 0.05 mmol.min-1.kg wet wt-1; P < 0.05) and coincided with a greater intramuscular acetyl-CoA-to CoASH ratio, acetyl-CoA content, and acetylcarnitine content. PDHa increased during exercise in both conditions but at a lower rate in the LCD condition compared with the HCD condition (1.46 +/- 0.25 vs. 2.65 +/- 0.23 mmol.min-1.kg wet wt-1 at 16 min and 1.88 +/- 0.20 vs. 3.11 +/- 0.14 at the end of exercise; P < 0.05). During exercise muscle acetyl-CoA and acetylcarnitine content and the acetyl-CoA-to-CoASH ratio decreased in the LCD condition but increased in the HCD condition. Under resting conditions PDHa was influenced by the availability of fat or carbohydrate fuels acting through changes in the acetyl-CoA-to-CoASH ratio. However, during exercise the activation of PDHa occurred independent of changes in the acetyl-CoA-to-CoASH ratio, suggesting that other factors are more important. PMID- 8238503 TI - Quantitation of glycolysis and skeletal muscle glycogen synthesis in humans. AB - We measured the net rates of skeletal muscle glycogen synthesis and glycolysis (conversion of [3-3H]glucose to 3H2O) in healthy overnight-fasted volunteers. Two studies were performed. In study 1, seven subjects participated in two paired infusions under basal conditions of either [2-3H]glucose (H2) or [3-3H]glucose (H3). Total glucose uptake (Rd) and rates of whole body 3H2O formation (3H2O Ra) were measured. With H2, Rd and 3H2O Ra were similar. With H3, 3H2O Ra, equal to glycolysis, was 65% of Rd. In study 2, six different subjects underwent a 3-h, 40 mU.m-2 x min-1 euglycemic insulin clamp. [6,6-2H2]glucose was infused throughout and H3 was infused during the last hour of the study. Open muscle biopsies were obtained at 150 and 180 min. Glycogen synthesis was assessed by three independent means: 1) direct measurement, as 3H disintegrations per minute in isolated muscle glycogen per plasma H3 specific activity; 2) extrapolation from the activity of glycogen synthase assayed in the presence of the concentrations of glucose 6 phosphate and UDP-glucose measured in the biopsy; and 3) the difference between Rd and glycolysis. Despite a wide range in Rd [24.5-58.8 mumol.kg fat-free mass (FFM)-1 x min-1] and glycolysis (14.2-26.1), the three methods yielded similar results of 20.0 +/- 3.9, 22.5 +/- 3.7, and 20.6 +/- 3.7 mumol.kg FFM-1 x min-1 and correlated highly with each other (r2 = 0.92-0.96). Our results (study 1) indicate that the rate of plasma tritiated water formation reflects the intracellular detritiation of tritiated glucose. Under hyperinsulinemic conditions (study 2) the net rate of muscle glycogen synthesis can be accurately estimated from the glycogen synthase activity and from the difference between total glucose uptake and glycolysis. Thus, at high physiological plasma insulin concentrations resulting in submaximal stimulation of muscle glycogen synthesis, the latter can be accurately measured in humans. PMID- 8238504 TI - IGF-I and pamidronate increase bone mineral density in ovariectomized adult rats. AB - Effects induced by insulin-like growth factor I (IGF-I) and/or the bisphosphonate pamidronate (APD) on bone mineral density (BMD) of the lumbar spine and proximal and midshaft tibia were studied in adult rats made osteopenic by ovariectomy, using dual-energy X-ray absorptiometry. IGF-I, which was administered by osmotic minipumps implanted subcutaneously for 6 wk, caused a dose-dependent increase of BMD at the three investigated sites. A 4-wk course of IGF-I, followed by intermittent cyclical APD administration, induced significant increases of BMD at the levels of spine and proximal tibia. At midshaft tibia, where cortical bone predominates, BMD was increased by IGF-I only. In conclusion, IGF-I increased BMD at sites with trabecular and/or cortical bone, whereas the APD influence was mainly detectable in the former site only. PMID- 8238505 TI - Correlation of diacylglycerol level and protein kinase C activity in rat retina to retinal circulation. AB - The increases in diacylglycerol (DAG) level and protein kinase C (PKC) activity have been characterized biochemically and functionally in the retina and the brain of diabetic rats as well as in cultured vascular cells. PKC specific activities were increased in the membraneous fraction of retina from streptozotocin (STZ)-induced diabetic rats and the genetically determined diabetic BB rats, respectively, after 1 or 2 wk of diabetes, compared with control. The ratio of total PKC activities from membraneous and cytosol fractions was also increased in the retina of diabetic rats. With diabetes, all the isoenzymes and the total DAG level were increased in the rat retina, whereas no changes were found in the rat brain. Insulin treatment normalized plasma glucose levels and partially prevented the increases in the membraneous PKC activity and all the isoenzymes in the retina. In the retinal endothelial cells, the total DAG level and PKC specific activities are increased by 36 and 22%, respectively, in the membraneous pool when the glucose levels are changed from 5.5 to 22 mM. Activation of PKC activity and isoform beta II by the vitreal injection of phorbol dibutyrate mimicked the abnormal retinal blood circulation observed in diabetic rats (2.22 +/- 0.24 vs. 1.83 +/- 0.40 s). Thus diabetes and elevated glucose levels will increase DAG level and PKC activities and its isoenzyme specifically in vascular cells and may affect retinal hemodynamics. PMID- 8238506 TI - Progressive alterations in lipid and glucose metabolism during short-term fasting in young adult men. AB - Stable isotope tracers and indirect calorimetry were used to evaluate the progressive alterations in lipid and glucose metabolism after 12, 18, 24, 30, 42, 54, and 72 h of fasting in six healthy male volunteers. The rates of appearance (Ra) of glycerol and palmitic acid in plasma doubled from 2.08 +/- 0.22 and 1.63 +/- 0.20 mumol.kg-1 x min-1, respectively, after 12 h to 4.36 +/- 0.36 and 3.26 +/- 0.40 mumol.kg-1 x min-1, respectively, after 72 h of fasting (P < 0.01). Of the total increase in lipid kinetics, 60% occurred between 12 and 24 h of fasting; the greatest interval change occurred between 18 and 24 h of fasting. Glucose Ra and plasma concentration decreased by approximately 25% between 12 h (11.0 +/- 0.4 mumol.kg-1 x min-1 and 5.58 +/- 0.08 mmol/l, respectively) and 72 h (8.3 +/- 0.3 mumol.kg-1 x min-1 and 4.14 +/- 0.10 mmol/l, respectively) of fasting (P < 0.01), but no statistically significant changes occurred between 18 and 24 h of fasting. Plasma insulin decreased by approximately 50% between 12 h (64.6 +/- 12.9 pmol/l) and 72 h (30.1 +/- 7.9 pmol/l) of fasting (P < 0.001). Of the total decline in plasma insulin, 70% occurred within the first 24 h of fasting. These results demonstrate that the mobilization of adipose tissue triglycerides increases markedly between 18 and 24 h of fasting in young adult men. The early alterations in lipid metabolism are associated with a decline in circulating insulin but do not seem to be regulated by changes in glucose kinetics or plasma glucose concentrations. PMID- 8238507 TI - Insulin resistance in the GK rat: decreased receptor number but normal kinase activity in liver. AB - We have previously shown that the glucose intolerance and the hyperglycemic state in the GK rat, a new spontaneous model of non-insulin-dependent (type II) diabetes without obesity, are partly accounted for by an alteration of the pancreatic B cell response. On the other hand, the hyperglycemic-hyperinsulinemic pattern in these rats suggests a decrease of response to insulin in the basal state. In the present study, in vivo insulin action was assessed in 8-wk-old GK females at basal and submaximal (euglycemic clamp) insulin levels. Overall glucose utilization (OGU), individual tissue glucose utilization (ITGU, in vivo uptake of the glucose analogue 2-deoxy-D-glucose as the relative index of glucose metabolism), as well as hepatic glucose production (GP) and liver insulin receptor properties were determined under these two conditions. The basal OGU was significantly higher in the GK females, compared with that in control Wistar females. The hyperinsulinemic-euglycemic clamp experiments indicated that peripheral insulin resistance was installed at 8 wk of age in the GK females because 1) OGU was significantly lower and 2) in some peripheral tissues (epitrochlearis muscle, periovarian, and inguinal white adipose tissues), but not all, ITGU was significantly lower compared with corresponding ITGU in control rats. In the basal state GP was significantly higher in the GK rats. At submaximal hyperinsulinemia (and euglycemia), it was less effectively suppressed than in the controls, thus demonstrating liver insulin resistance. Under both basal state and clamp condition, binding of 125I-A14-insulin to liver membranes of GK rats was significantly decreased by 20-30%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238508 TI - Model for measuring absolute rates of hepatic de novo lipogenesis and reesterification of free fatty acids. AB - We have previously presented a precursor-product stable isotopic technique for measuring in vivo the fraction of very low-density lipoprotein-fatty acids (VLDL FA) derived from de novo lipogenesis (fractional DNL). Here, we propose a technique for converting fractional DNL into absolute rates of DNL and describe its explicit underlying assumptions. The technique combines the fractional DNL method with a modification of the method of S. Klein, V. R. Young, G. L. A. Blackburn, B. R. Bistrain, and R. R. Wolfe (J. Clin. Invest. 78: 928-933, 1986), for estimating hepatic reesterification of free fatty acids (FFA). Infusions of [1,2,3,4-13C]palmitate and [1-13C]acetate are performed concurrently with indirect calorimetry in human subjects. Fractional DNL (based on mass isotopomer distribution analysis of VLDL-FA), the rate of appearance of plasma FFA (Ra of FFA), and net fat oxidation in the whole body are measured. Equations from the hepatic reesterification model, modified to include the contribution from DNL, allow calculation of absolute DNL (= fractional DNL x [Ra of FFA - net whole body fat oxidation], when respiratory quotient < 1.0). Sample results from human subjects with different dietary energy intakes are presented, with calculations of absolute DNL, absolute reesterification, and absolute fat oxidation rates. The assumptions of this technique (in particular, that all fat oxidized is derived at steady state from circulating FFA and that DNL and reesterification of FFA both occur exclusively in liver) are discussed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238509 TI - Identification of cholinergic neurons in enteric nervous system by antibodies against choline acetyltransferase. AB - Several different monoclonal and polyclonal antibodies to choline acetyltransferase (ChAT) were screened to identify effective antibodies for immunocytochemical marking of cholinergic neurons in the enteric nervous system. Excellent immunohistochemical results were obtained with two of the antibodies in the myenteric plexus of the guinea pig stomach and small intestine. One was a mouse monoclonal antibody designated B3.9B3, and the second was a rabbit polyclonal antibody referred to as Peptide 3. Both antibodies clearly stained neuronal cell bodies as well as nerve fibers to the muscle layers and fibers encircling stained and unstained cell bodies. Cell counts indicated that approximately 64% (21.0 +/- 8.6 cells/ganglion) of gastric myenteric neurons are ChAT positive. Pelvic ganglia and the inferior mesenteric ganglia were examined as controls. Strong labeling of the majority of neurons was found in the pelvic ganglia, whereas few immunoreactive cells were apparent in the predominantly noradrenergic inferior mesenteric ganglion. Lack of effective antibodies to enteric neuronal ChAT has hampered progress in the study of the neurophysiology of cholinergic neurons in the digestive tract. Application of the B3.9B3 and Peptide 3 antibodies now promises to facilitate investigation of this important subset of enteric neurons. PMID- 8238510 TI - Effect of [(CH2NH)4,5]secretin on pancreatic exocrine secretion in guinea pigs and rats. AB - [psi 4,5]Secretin was shown to be a secretin receptor antagonist that inhibits secretin-stimulated increase in adenosine 3',5'-cyclic monophosphate in isolated pancreatic acini of the guinea pig. To determine whether it inhibits pancreatic exocrine secretion in vivo, we have studied the effect of [psi 4,5]secretin on the pancreatic secretion stimulated by secretin in anesthetized guinea pigs and rats. In basal state, [psi 4,5]secretin given intravenously for 2 or 3 h in varying doses of 1.6-32.7 nmol.kg-1.h-1 dose dependently increased pancreatic secretion of both fluid and bicarbonate during the 1st h, but it returned gradually to basal level within 2 or 3 h. On the other hand, [psi 4,5]secretin significantly inhibited the pancreatic secretion stimulated by either exogenous or endogenous secretin in a dose-related manner. The inhibitory effect of [psi 4,5]secretin in guinea pigs was greater than that in rats. However, it did not completely block the secretin-stimulated pancreatic secretion, whereas a rabbit antisecretin serum suppressed it completely. We conclude that 1) in the unstimulated state, [psi 4,5]secretin is a partial agonist of pancreatic exocrine secretion of both fluid and bicarbonate; and 2) when pancreatic secretion is stimulated by secretin, unlike an antisecretin serum, it is a partial inhibitor in intact guinea pigs and rats. PMID- 8238511 TI - Structural characterization of PACAP receptors on rat liver plasma membranes. AB - Pituitary adenylate cyclase-activating polypeptide-38 (PACAP-38) and PACAP-27 are recently characterized hypothalamic peptides with marked homology with vasoactive intestinal peptide (VIP), which are concentrated in the innervation of the digestive tract. We now report that, on rat liver plasma membranes, PACAP interacts with at least two types of receptors: receptors demonstrating equally high affinity for PACAP and VIP and receptors with high affinity for PACAP but low affinity for VIP. In contrast, on rat intestinal epithelial cell laterobasal membranes, only receptors with high affinities for PACAP and VIP were observed. After 125I-labeled VIP or 125I-labeled PACAP-27 was cross-linked to the liver plasma membrane receptors with the use of either disuccinimidosuberate or disuccinimido dithiobis(propionate), analysis of the resulting ligand-receptor complexes on sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the structures of the VIP and PACAP receptors were similar: both ligand receptor complexes displayed two radioactive bands with relative molecular weights of 80,000 and 56,000 under reducing conditions and of 75,000 and 53,000 under nonreducing conditions. These findings suggest that the receptors for the PACAP peptides and VIP are closely related, reflecting the marked homology between these peptides. The presence of receptors specific for PACAP on rat liver plasma membranes should stimulate further studies of the interaction between PACAP and the liver. PMID- 8238512 TI - Sex differences in intracellular fatty acid transport: role of cytoplasmic binding proteins. AB - Female liver clears long-chain fatty acids from plasma more rapidly than male liver, and yet the basis for this sex difference is poorly understood. We tested the hypothesis that cytosolic fatty acid binding protein (FABP), which is more concentrated in female liver, may enhance fatty acid utilization by increasing the rate of transport through the cytoplasm. We modified the technique of fluorescence recovery after laser photobleaching to measure the cytoplasmic diffusion rate of the fluorescent long-chain fatty acid 12-N-methyl-(7-nitrobenz 2-oxa-1,3-diazol)aminostearate (NBD-stearate) in cultured hepatocytes from female and male rats. NBD-stearate was used because its hepatic handling is similar to natural fatty acids. After uptake, NBD-stearate distributed uniformly in the cytoplasm but was excluded from the nucleus. Intracellular transport occurred by diffusion with no detectable convective flux. The cytoplasmic diffusion rate at 37 degrees C was 65% greater in female cells than in male cells (mean +/- SE, 5.03 +/- 0.37 vs. 3.05 +/- 0.21 x 10(-9) cm2/s respectively; P < 0.001) and was two to three orders of magnitude slower than for either unbound NBD-stearate or FABP in water. A correspondingly greater fraction of cellular NBD-stearate was found in the aqueous cytosol in females (35.1 +/- 7.0 vs. 18.2 +/- 2.7%), suggesting that FABP reduces binding of NBD-stearate to immobile cytoplasmic membranes. These data indicate that intracellular transport of NBD-stearate, a typical amphipathic molecule, is slowed by binding to cytoplasmic membranes. The primary function of soluble binding proteins such as FABP may be to enhance the diffusive fluxes of their ligands by reducing membrane binding. If cytoplasmic transport of rapidly metabolized fatty acids such as palmitate is similarly slow, substantial concentration gradients could develop within the cytoplasm of hepatocytes at steady state. By catalyzing these diffusive fluxes, FABP may regulate fatty acid metabolism. PMID- 8238513 TI - Characterization of univectorial polyamine transport in duodenal crypt cell line. AB - High levels of polyamines have been identified in the lumen of the intestines. Luminal polyamines are involved in normal mucosal growth and may be the primary source of extracellular polyamines for tumors grown in animals undergoing polyamine antimetabolite therapy. The vectorial movement of polyamines across an in vitro model of the gut was studied in epithelial cells grown in culture. IEC-6 cells were plated on either plastic or raised inserts. Cells grown on plastic were employed to define the kinetic constants for putrescine and spermidine uptake. Eadie-Hofstee plot analysis of putrescine uptake was characteristic of a single class of transporter with a Michaelis constant (Km) of 4.85 +/- 0.57 microM and a maximal velocity (Vmax) of 627 +/- 85 pmol x 15 min-1 x 10(6) cells 1. The plot for spermidine uptake was curvilinear and representative of the interaction of spermidine with two sites: Km 1 and 2 are 0.26 +/- 0.13 and 2.1 +/ 0.77 microM; Vmax 1 and 2 are 177 +/- 50 and 429.5 +/- 70 pmol x 15 min-1 x 10(6) cells-1, respectively. Calmodulin antagonism blocked the uptake of putrescine and the low-affinity but not high-affinity spermidine uptake system. Seven-day postconfluent cells grown on plastic inserts were used to study the vectorial movement of polyamines in a polarized epithelium. The apical membrane domain expressed two sites with similar kinetic constants to those observed when cells were grown on plastic. In contrast, however, the basolateral membrane did not transport polyamines. Spermidine uptake through this membrane was only a fraction of that in the apical membrane and was completely nonspecific.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238514 TI - Cyclin and cyclin-dependent kinase 1 mRNA expression in models of regenerating liver and human liver diseases. AB - There is compelling evidence that the eukaryotic cell cycle is controlled by a family of proteins called cyclins, which complex with cyclin-dependent kinases (CDK) to modulate key events during cell division. We have examined the regulation of these genes in models of experimental liver regeneration and their expression in human liver diseases. Seventy percent partial hepatectomy (PH) was performed on rats and normal BALB/c and athymic nude mice to determine patterns of cyclin and CDK1 mRNA expression. It has been previously shown by [3H]thymidine incorporation that athymic nude mice manifest impaired regeneration after PH. Our results demonstrate a sequential pattern of cyclin and CDK1 transcript expression in each of the models. Cyclin D1 was the most abundant mRNA steady-state transcript in the regenerating livers. CDK1 and cyclins associated with later stages of the cell cycle showed delayed and diminished expression in nude mice compared with normals. Nuclear run-off assays performed at key time points post PH revealed little change in transcription rates, suggesting that steady-state mRNA expression of the cyclin genes is regulated primarily by posttranscriptional events. Human liver tissue from various acute and chronic hepatic diseases showed increased expression of cyclins A and D1. We conclude that the regenerating liver post-PH offers an excellent in vivo model for studying cyclin and CDK gene expression. Impaired regeneration in the nude mouse is associated with altered cyclin and CDK1 mRNA transcript expression. Furthermore, cyclins may eventually provide clinically relevant molecular markers of regenerative activity in human liver diseases. PMID- 8238515 TI - Partial agonism by gastrin for a cholecystokinin receptor mediating pepsinogen secretion. AB - Isolated gastric glands from rabbit were used to characterize the functional cholecystokinin (CCK)-like peptide receptors that mediate pepsinogen secretion. Pepsinogen secretion was stimulated by both CCK octapeptide sulfate (CCK-8) and A 71378, a selective CCK-A-type receptor agonist, with similar mean effective doses (1.0 and 0.8 nM, respectively). Compared with CCK-8, gastrin-17 (G-17-I) showed reduced potency and only partial efficacy for stimulation of pepsinogen secretion while inhibiting the maximal CCK-8-stimulated response. The nonpeptide inhibitors, asperlicin and L-364,718, inhibited pepsinogen secretion with identical pA2 values for antagonism of both CCK and gastrin, indicating that both peptides interact with the same functional receptor. Specific binding of [3H]CCK 8 to isolated chief cell membranes was displaced fully by both CCK and gastrin, indicating full receptor occupancy by both peptides. A novel synthetic peptide analogue, pseudogastrin [(Glu)5-Ala-Tyr-Nle-Gly-Trp-Nle-Asp-Phe-NH2], was used to investigate the structural basis for the lower potency and efficacy of G-17-I. The potency of CCK and gastrin analogues for pepsinogen secretion was found to be dependent on both sulfation of a tyrosine residue and the position of the tyrosine residue relative to the COOH-terminal phenylalanine amide. The efficacy appears to be determined partially by the extended NH2-terminal sequence of G-17 I. The results of the present study are interpreted to show that pepsinogen secretion is mediated by a CCK-A-type receptor and gastrin acts at the same receptor as a partial agonist. PMID- 8238516 TI - Potassium current in circular smooth muscle of human jejunum activated by fenamates. AB - Thirty-eight cells, freshly isolated from circular smooth muscle of normal human jejunum obtained from nine patients undergoing weight-reduction surgery for morbid obesity, were patch clamped using a perforated patch-clamp technique. A highly potassium-selective voltage-dependent outward current was present in all cells. The current was carried by a 220-pS channel that activated near -75 mV and reached unit open probability at about +10 mV. Blockade of the current by quinidine (50 microM) and tetraethylammonium (25 mM) was accompanied by membrane depolarization to 0 to -3 mV, suggesting that this current was the major determinant of the membrane potential. Flufenamic and mefenamic acid at concentrations comparable with blood levels reached when these drugs are used in clinical therapy as nonsteroidal anti-inflammatory agents, activated the potassium outward current and hyperpolarized the membrane potential. The shift in the membrane potential for 250 microM flufenamic acid was -36 +/- 24 (SD) mV. Activation was rapid (seconds) and reversible. It was concluded that normal human jejunal circular smooth muscle cells have a highly potassium-selective outward current, which is the major determinant of the membrane potential and which is activated by fenamates. PMID- 8238517 TI - TRH in dorsal vagal complex mediates acid response to excitation of raphe pallidus neurons in rats. AB - The role of thyrotropin-releasing hormone (TRH) in the dorsal vagal complex (DVC) in the acid response to excitation of raphe pallidus neurons was investigated in urethan-anesthetized rats with gastric fistula. Kainic acid (0.19 microgram/30 nl) microinjected into the raphe pallidus stimulated gastric acid secretion. The response was prevented by vagotomy. A specific polyclonal TRH antibody, 8964, was raised and characterized (50% inhibitory dose for TRH was 80 pg/ml at an antibody final dilution of 1:10(5)). The TRH antibody injected intracisternally blocked the acid response to intracisternal TRH, but not that of the TRH analogue RX 77368. The TRH antibody (0.33, 0.65, or 1.3 micrograms.100 nl-1.site-1) microinjected bilaterally into the DVC prevented dose dependently by 31, 60, and 76%, respectively, the increase in acid secretion induced by kainic acid injected into the raphe pallidus. The TRH antibody (1.3 microgram/site) microinjected into medullary sites outside of the DVC had no effect. These data indicate that excitation of raphe pallidus neurons induces a vagal-dependent stimulation of gastric acid secretion that is mediated by endogenous TRH in the DVC. TRH neurons in the raphe pallidus projecting to the DVC may have a physiological relevance in the vagal regulation of gastric function. PMID- 8238518 TI - Actions of nitric oxide-generating sodium nitroprusside in myenteric plexus of guinea pig small intestine. AB - Sodium nitroprusside (NaNP) was used as a donor of nitric oxide (NO) to investigate actions of NO on electrical and synaptic behavior of single myenteric neurons in guinea pig small intestine. NaNP (10 microM-1 mM) did not affect resting membrane properties of the neurons, except for an occasional decrease in input resistance and hyperpolarization attributable to suppression of excitatory transmitter release. NaNP did not alter fast nicotinic neurotransmission but suppressed noncholinergic slow excitatory postsynaptic potentials (slow EPSPs) in a concentration-dependent manner. Pretreatment with either methylene blue or oxyhemoglobin reduced the inhibitory action of NaNP on the slow EPSPs. Slow EPSP like responses to microejected substance P or 5-hydroxytryptamine were unaffected by NaNP. The nitric oxide synthase inhibitor, N omega-nitro-L-arginine methyl ester, did not affect resting membrane excitability or excitatory synaptic events in any of the myenteric neurons. The results suggest that NO may not be released extensively as a neurotransmitter at synapses within the myenteric plexus. If myenteric neurons are exposed to NO released from nonneural sources, then the principal action is expected to be presynaptic inhibition of slow synaptic excitation. PMID- 8238519 TI - Comparison of plasma membrane FABP and mitochondrial isoform of aspartate aminotransferase from rat liver. AB - A relationship between plasma membrane fatty acid binding protein (FABPpm), a putative membrane transporter for long-chain fatty acids, and the mitochondrial isoform of aspartate aminotransferase (m-AspAT) has been reported. Accordingly, we have compared the chemical and immunological properties of rat liver m-AspAT with those of rat liver FABPpm isolated by two procedures: 1) detergent solubilization of the membranes followed by purification via fatty acid affinity chromatography (FABP-1) or 2) salt extraction of the membranes and subsequent purification by high-performance liquid chromatography (HPLC; FABP-2). Comparison of the three protein preparations revealed no differences with respect to NH2 terminal amino acid sequence, amino acid composition, peptides from tryptic digests, AspAT enzymatic activity, isoelectric point, mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), retention on five different HPLC columns, and immunoprecipitation and immunoblotting of SDS-PAGE separated proteins with polyclonal antisera. Examination of the proteins by nondenaturing PAGE showed a consistent second band in FABP-1 and FABP-2 not always present in m-AspAT. However, whenever present, this band was immunoreactive with antibodies to both m-AspAT and FABP-1. Hence, FABP-1 and FABP 2 are indistinguishable from one another. They are also at least closely related, if not identical, to m-AspAT. PMID- 8238520 TI - Intracellular pH-regulating ion transport mechanisms in parietal cell basolateral membrane vesicles. AB - Na(+)-H+ and Cl(-)-base exchangers on the parietal cell have been demonstrated by several authors. Controversy exists concerning a basolateral Na(+)-HCO3- cotransporter in the parietal cell. To clarify this issue, we prepared highly enriched basolateral membrane (BLM) and apical-tubulovesicular membrane (to serve as negative controls) vesicles from rabbit fundic mucosa. Na(+)-H+ exchange was demonstrated by measuring pH gradient-driven amiloride-sensitive 22Na+ uptake and Na+ gradient-driven proton uptake into voltage-clamped BLM but not into apical tubulovesicular vesicles. Anion exchange was demonstrated by measuring 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS)-inhibitable influx of 36Cl- into Cl(-)- or HCO3(-)-loaded voltage-clamped BLM vesicles. Na(+)-HCO3- cotransport was assessed by comparing HCO3(-)-driven 22Na uptake with uptake driven by an identical pH gradient. No significant difference was found between 22Na uptake in the presence and absence of HCO3-; 1 mM amiloride inhibited 22Na uptake > 90% in both conditions, whereas 2 mM DIDS had no effect. In BLM vesicles prepared from rabbit renal cortex, however, a HCO3- gradient stimulated 22Na uptake much more than an equivalent pH gradient, and DIDS inhibited this HCO3- gradient-driven 22Na uptake. This indicates that our experimental setup was suitable to detect a Na(+)-HCO3- cotransporter if present. Our data suggest that the parietal cell BLM contains Na(+)-H+ exchangers and Cl(-)-HCO3- exchangers but no Na(+)-HCO3- cotransporter. PMID- 8238521 TI - Expression of gap junction connexin 43 and connexin 43 mRNA in different regional tissues of intestine in dog. AB - To determine the distribution of gap junction protein connexin 43 (Cx43) and the expression of Cx43 mRNA in different regions of canine small intestine and colon, modified Western blot and Northern blot techniques were used with a site-directed antibody raised against a synthetic peptide of Cx43 Anti-Cx43 (252-271) and a probe of 1.3-kb Cx43 cDNA. Equal amounts of plasma membrane enriched fraction (10 micrograms protein) from small intestinal and colonic circular muscle, longitudinal muscle, and colonic submucous plexus border of circular muscle (interstitial cells of Cajal rich layer, ICC) of the dog were resolved by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred onto nitrocellulose, and blotted by chemiluminescent assay. Different yet characteristic ratios of the immunoreactive gap junction proteins located at 43 and 40 kDa were found in membranes from small intestinal and colonic circular smooth muscle, and ICC. These results suggest that gap junction Cx43 in dog heart, dog small intestinal and colonic circular muscle, and ICC is homologous to rat heart. Furthermore, with the use of Northern blot hybridization with a 1.3-kb Cx43 cDNA, a 3.0-kb message was observed in small intestinal and colonic circular muscles, longitudinal muscles, and ICC. However, the mRNA signal of small intestinal circular muscle was the strongest and that of longitudinal muscle was weakest, especially from colon longitudinal muscle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238522 TI - Characterization of bumetanide transport in isolated skate hepatocytes. AB - The uptake of [3H]bumetanide was studied in isolated skate hepatocytes in an albumin-free elasmobranch Ringer solution and compared with the uptake of bile acids in the presence of other cholephilic organic anions. [3H]bumetanide uptake was energy dependent, temperature sensitive, and exhibited saturation kinetics. In contrast to taurocholate and cholate, which are transported only by Na(+) independent mechanisms, removal of Na+ reduced the maximal uptake rate (Vmax) for bumetanide from 404 +/- 80 to 230 +/- 47 pmol.mg-1 x min-1 without a change in the apparent Michaelis constant (Km). The apparent Km for the Na(+)-dependent portion of bumetanide uptake was 58 +/- 24 microM, and Vmax was 151 +/- 38 pmol.min-1 x mg-1. Taurocholate (100 and 200 microM) inhibited Na(+)-independent bumetanide transport competitively but was a noncompetitive inhibitor for Na(+) dependent bumetanide uptake. Furosemide (100 microM) and two bumetanide analogues, PF-3034 (500 microM) and PF-2203 (500 microM), preferentially inhibited the Na(+)-dependent bumetanide uptake system, whereas cholate (100 microM) and probenecid (100 microM) preferentially inhibited Na(+)-independent bumetanide transport. The sulfhydryl (SH) reagents N-ethylmaleimide, 2,2'-dithio bis(5-nitropyridine), and p-chloromercuribenzenesulfonic acid (PCMBS) inhibited both bile acid and bumetanide uptake. Dithiothreitol (500 microM) completely reversed the PCMBS-induced inhibition of bumetanide uptake. These results indicate that bumetanide is transported into hepatocytes of the small skate, Raja erinacea, by both Na(+)-dependent and Na(+)-independent mechanisms; the latter is shared by bile acids and probably sulfobromophthalein and other organic anions. Their uptake requires free SH groups. PMID- 8238523 TI - Nonparallel secretion of pepsinogen and acid by gastric oxyntopeptic cells of the toad (Bufo marinus). AB - Pepsinogen and HCl secretion in the amphibian stomach are performed by a single cell type, the oxyntopeptic cell. These functions were studied in gastric mucosae of toads (Bufo marinus) mounted in Ussing-type chambers. HCl and peptic activity of luminal fluid were measured by titration and proteolysis of albumin, respectively. Distribution of pepsinogen in the gastric mucosa was heterogeneous, activity being highest in the proximal part of the stomach. Zymogen granules in the oxyntopeptic cell were more abundant in the deeper cells of the glands and in the fundus. On stimulation, the granules were released into the lumen of the glands by exocytosis. Histamine, forskolin, or carbachol alone each induced an increase in HCl and pepsinogen secretion. Carbachol after maximal histamine or forskolin stimulation produced an extra increase in both secretions that was greater for pepsinogen response. Similarly, joint addition of carbachol and histamine was more potent than histamine alone for both parameters; however, the effect was greater on pepsinogen release. Pretreatment with cimetidine blocked HCl and pepsinogen responses to carbachol but did not affect responses to forskolin. Addition of omeprazole to forskolin-stimulated mucosae uncoupled the two secretions, inducing a total inhibition of HCl secretion with a slight reduction in pepsinogen secretion. Thus pepsinogen release, similar to HCl secretion, is sensitive to cAMP and Ca(2+)-dependent secretagogues. However, the action of Ca2+ would require the previous elevation of cAMP induced by the different secretagogues. In such a case, the increase in intracellular Ca2+ would result in a nonparallel activation of the two secretions. PMID- 8238524 TI - Interaction of bumetanide derivatives with hepatocellular bile acid uptake. AB - The loop diuretic bumetanide is an organic monocarboxylic organic anion assumed to be transported into hepatocytes by a transport system for bile acids. The structural requirements of 22 bumetanide analogues were analyzed for an interaction with bile acid uptake into isolated rat hepatocytes. Whereas bumetanide inhibited the hepatocellular uptake of [14C]cholate to the same degree as its own uptake, derivatization altered affinity and specificity and yielded compounds that selectively inhibited either cholate or taurocholate uptake or uptake of both. No correlation was found between the diuretic potency of bumetanide derivatives, reflecting the affinity to the Na(+)-K(+)-Cl- cotransporter, and their affinity to hepatic bile salt transport. Computer-aided model building combined with the calculation of potential energy maps showed a strictly amphipathic charge separation in bumetanide analogues as in bile acids. Ranking bumetanide compounds by their mean inhibitory concentration values, inhibition constants, and their type of competition, we conclude that at least three binding domains in the proteins are essential for recognition by bile acid transporters, namely two hydrophobic and an anionic side, and that for the anionic binding region a carbonyl atom in the ligands as an electron donor group is sufficient for ligand interaction. PMID- 8238525 TI - Regulation of paracellular permeability in Caco-2 cell monolayers by protein kinase C. AB - Caco-2 cells are an enterocyte-like cell line derived from a human colonic adenocarcinoma. Paracellular permeability was assessed in monolayers of these cells by transmonolayer resistance and by the permeation of [3H]mannitol across the monolayer. Paracellular permeability was increased by the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (50 nM), carbachol (500 microM), and the combination of carbachol (50 microM) and monolein (100 microM), an inhibitor of diacylglycerol kinase, as manifested by a decrease in transmonolayer resistance and an increase in mannitol permeation. The effects of all of these stimuli on transmonolayer resistance were inhibited by staurosporine (3 nM), an inhibitor of PKC. The effects of carbachol plus monolein were also inhibited by atropine (0.1 microM), a muscarinic antagonist. Treatment of the monolayers with each of the stimuli was associated with translocation of PKC activity from cytosol to a membrane-associated state. Stimulation of Caco-2 cell monolayers with phorbol myristate acetate or with the combination of carbachol and monolein was also associated with phosphorylation of the MARCKS protein, an endogenous substrate of PKC. These data support the hypothesis that intestinal paracellular permeability is regulated by the activity of enterocyte PKC and demonstrate that the increase in paracellular permeability induced by binding of carbachol to the muscarinic receptor is mediated by activation of PKC. PMID- 8238526 TI - Coordination of Cl- secretion and contraction by a histamine H2-receptor agonist in guinea pig distal colon. AB - Short-circuit current (Isc) was measured simultaneously with muscle tension recorded in the longitudinal or circumferential axis from strain-gauge transducers sutured to the serosal surface of whole thickness segments of distal colon from guinea pigs. Isc and the amplitude and frequency of small-amplitude phasic contractions were stable for several hours. The histamine H2-receptor agonist dimaprit (5-10 microM) evoked recurrent increases in Isc at a frequency of 0.32-0.37 cycles/min. Associated with each cyclical Isc response was an increase in muscle tension indicative of large-amplitude phasic contractions. Recurrent cycles of Isc and associated large-amplitude phasic contractions were abolished by 10 microM cimetidine. In the presence of dimaprit, severing the neural connections between the myenteric and submucosal plexuses abolished the large-amplitude phasic contractions but not cyclical Isc. The results suggest that coordination of cyclical secretion and large-amplitude contraction during chronic activation of histamine H2 receptors requires intact neural connections between the submucosal and myenteric plexuses. The findings provide potential mechanisms to explain altered motility and ion transport during inflammatory diseases or allergic responses to food antigens. PMID- 8238527 TI - Cotransport of 2-methyl-aminoisobutyric acid and chloride in rabbit small intestine. AB - This article examines the nature of the Cl- dependence of the rabbit intestinal imino acid carrier using 2-methyl-aminoisobutyric acid (MeAIB) as substrate for measuring influx across the luminal membrane of rabbit distal ileum. Reduction of mucosal Cl- concentration from 140 to 6 mM successively increased the concentration at which MeAIB influx is half-maximal from 3.6 +/- 0.2 (SD) to 21.0 +/- 3.7 mM. Maximal influx of MeAIB was unchanged. Measured at 70 mM Cl-, MeAIB induced a Cl- influx across the brush-border membrane (delta JClmc) of 5.01 +/- 0.65 mumol.cm-2 x h-1 (SE, n = 16; 45%), and an identical Cl(-)-independent influx of leucine induced a delta JClmc of 1.82 +/- 0.52 mumol.cm-2 x h-1 (SE, n = 12; 16%) (P < 0.001). At 24 mM Cl-, delta JClmc induced by leucine did not differ significantly from zero, whereas the effect of MeAIB was maintained. The ratio between Na+ influx across brush-border membrane (delta JNamc) and the Na(+) dependent flux measured at 140 mM Cl- was 2.4 +/- 0.7 (n = 8) for MeAIB, 1.4 +/- 0.4 (n = 10) for leucine, and 1.1 +/- 0.2 (n = 13) for D-glucose. The Hill coefficient of Na+ activation (+/- SD) in distal ileum was 0.82 +/- 0.19 for leucine and 1.14 +/- 0.03 for D-glucose. A Hill coefficient of 1.60 +/- 0.05 was obtained for D-glucose in the jejunum. Concentrations of MeAIB and D-glucose, chosen to given equal delta JNamc, induced a 40 and 21% increment of JClmc (P < 0.05), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238528 TI - A cloned CCK-A receptor transduces multiple signals in response to full and partial agonists. AB - Chinese hamster ovary (CHO) cells, stably transfected with the cloned rat CCK-A receptor, were used to study signal transduction events initiated by cholecystokinin octapeptide (CCK-8) and the partial agonist JMV-180. In single CHO-CCK-A cells loaded with fura-2, superfusion of CCK-8 (10 pM-1 nM) resulted in an increase in intracellular Ca2+ concentration ([Ca2+]i). At CCK-8 concentrations < 100 pM, the signal consisted of [Ca2+]i oscillations. At higher concentrations, CCK-8 induced a typical biphasic response consisting of a large peak followed by a lower sustained plateau. Superfusion of JMV-180 also resulted in an increase in [Ca2+]i; in contrast to acinar cells this increase did not consist exclusively of [Ca2+]i oscillations. Both CCK-8 and JMV-180 increased polyphosphoinositide hydrolysis, although JMV-180 stimulated formation of only 10% as much [3H]inositol phosphates. [Ca2+]i signals stimulated by both CCK-8 and JMV-180 were blocked by the aminosteroid U-73122. CCK-8 (1-10 nM) increased formation of adenosine 3',5'-cyclic monophosphate (cAMP) and release of arachidonic acid in CHO-CCK cells. These increases were not mimicked by JMV-180 (10 microM). Furthermore, no cAMP formation or arachidonate release could be detected when cells were incubated with both JMV-180 and CCK-8. These data indicate that in CHO-CCK-A cells, unlike acinar cells, both CCK-8 and JMV-180 increase [Ca2+]i by similar mechanisms. However, the CCK-A receptor can differentially recognize and then activate discrete transduction pathways on binding of these two agonists. PMID- 8238529 TI - Radioligand binding methods: practical guide and tips. AB - Radioligand binding assays are a relatively simple but extremely powerful tool for studying receptors. They allow an analysis of the interactions of hormones, neurotransmitters, growth factors, and related drugs with the receptors, studies of receptor interactions with second messenger systems, and characterization of regulatory changes in receptor number, subcellular distribution, and physiological function. As a result, these assays are widely used (and often misused) by investigators in a variety of disciplines, including pharmacology, physiology, biochemistry, immunology, and cell biology. This article presents a broad overview of the radioligand binding assay technique, primarily for the investigator who has limited experience with this technique. Practical guidelines for setting up a new assay are presented, including the receptor preparation to be used, choice of appropriate radioligand, optimizing assay conditions, and appropriate methods for data analysis. Tips for avoiding some of the common pitfalls in application of these assays are also included. The primary focus is on radioligand binding assays of membrane-bound receptors studied in membrane preparations. However, similar assay techniques can be used to study receptors on intact cells. The unique advantages and disadvantages of these intact cell binding assays are also discussed. In particular, the occurrence of regulatory changes in receptors during the course of intact cell binding assays is considered, with approaches for circumventing these complications and for using intact cell assays to advantage in studying these regulatory changes. PMID- 8238530 TI - Retinoic acid increases elastin in neonatal rat lung fibroblast cultures. AB - The factors that regulate elastin synthesis during pulmonary alveolar septal formation have not been identified. Because maximal alveolar elastin synthesis occurs over a relatively brief period (postnatal days 4-14 in the rat), we hypothesized that changes in the local concentrations of factors that regulate elastin synthesis may precede or accompany this period. Because pulmonary retinoid stores decline just before the fourth postnatal day, we also hypothesized that this decline could be accompanied by the utilization of retinoic acid, one of the most biologically active retinoids, in a regulatory process that increases elastin synthesis. If these hypotheses are correct, then retinoic acid should increase elastin synthesis by pulmonary cells. Therefore, cultures of neonatal rat lung fibroblasts were exposed to retinoic acid, and elastin production was quantitated. Retinoic acid produced a two- to threefold increase in the steady-state level of elastin mRNA, in soluble elastin, and in insoluble elastin. The transcriptional initiation rate of the elastin gene was 1.8-fold higher in nuclei that were isolated from retinoic acid-treated cells than in nuclei that were isolated from control cells. This indicates that the increase in steady-state elastin mRNA results, at least partially, from an increase in elastin transcription. Lung fibroblasts that were isolated from 8-day old rats, but not cultured, contained retinoic acid. These findings suggest that retinoic acid is a potential regulator of elastin synthesis in developing pulmonary alveoli. PMID- 8238531 TI - A 96-kDa gelatinase induced by TNF-alpha contributes to increased microvascular endothelial permeability. AB - Tumor necrosis factor-alpha (TNF-alpha) may increase vascular endothelial permeability through alteration of the extracellular matrix (ECM). Incubation of bovine pulmonary microvascular endothelial (BPMVE) cells grown to confluence on microporous filters with 10(4) U/ml TNF-alpha for 24 h increased monolayer permeability to 125I-labeled albumin two- to threefold. TNF-alpha treatment also induced expression of a 96-kDa gelatinolytic metalloproteinase that was present in the medium and bound to the ECM. The induced 96-kDa metalloproteinase was purified from conditioned medium and found to cleave fibronectin, laminin, types IV and V collagens, and gelatins from types I and III collagens, suggesting identity as a type IV collagenase-gelatinase. Incubation of BPMVE cells with the 96-kDa gelatinase increased monolayer permeability, an effect prevented by inclusion of either tissue inhibitor of metalloproteinase (TIMP) or 1,10 phenanthroline. When BPMVE cells were incubated with the 96-kDa gelatinase or 10(4) U/ml TNF-alpha and then stripped from the filters, the remaining ECM displayed increased permeability to 125I-albumin compared with matrix from untreated BPMVE. The ECM extracts from both TNF-alpha- and enzyme-treated cells were found to contain less fibronectin, whereas their total protein contents were similar to those of untreated controls. These results suggest that the 96-kDa metalloproteinase induced by TNF-alpha contributes to increased vascular endothelial permeability through the degradation of specific extracellular matrix components. PMID- 8238532 TI - Degradation of surfactant protein B by alveolar type II cells. AB - Surfactant protein B (SP-B) metabolism was studied in primary cultures of alveolar type II cells. Iodinated SP-B reconstituted with surfactant was incorporated rapidly into lung pneumocytes and degraded to trichloroacetic acid (TCA)-soluble products after a lag period of 1 h. Cellular degradation of SP-B occurred whether or not phospholipid liposomes or surfactant was added to the phospholipid-poor SP-B. Uptake and degradation of SP-B at 37 degrees C showed a linear increase up to 3 micrograms SP-B/ml after which the slope of the curve became less steep with increasing concentrations of SP-B in the media. After 4 h of incubation with SP-B, 35% of the SP-B processed was recovered as degradation products. Ninety-six percent of the degradation products were in the media and only 4% were recovered in the cell. The bulk of the breakdown of SP-B occurred inside the type II cells since degradation did not occur at 4 degrees C, showed a 1-h lag period, was proportional to the SP-B protein internalized by the cells, was inhibited 47% by ammonium chloride, was unaffected by the addition of protease inhibitors to the medium, and cell-conditioned medium produced only limited SP-B degradation. Alveolar macrophages also degraded SP-B, whereas other cell types degraded SP-B to a lesser extent. Thus the specificity of the metabolism of SP-B may be through the capability of lung cells to degrade SP B.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238533 TI - Tumor necrosis factor-alpha alters pulmonary vasoreactivity via neutrophil derived oxidants. AB - We postulated that tumor necrosis factor-alpha (TNF) "primes" the lung for the development of pulmonary vasoconstriction and edema by inducing the release of polymorphonuclear leukocyte (PMN)-derived reactive oxidant species (ROS). Guinea pigs were injected with TNF (1.6 x 10(5) U/kg ip), and the lungs isolated 18 h later. Compared with controls, TNF pretreatment resulted in 1) greater increases in lung weight and capillary pressure in response to the thromboxane A2 mimetic U 46619 (365 pmol/min) and 2) an increase in the dose of acetylcholine (ACh) causing 50% of maximal dilation (EC50). The vascular effects of TNF were associated with 1) decreased lung effluent nitrite (NO2-, oxidation product of nitric oxide), 2) increased lung effluent superoxide (O2-), and 3) increased lung myeloperoxidase (MPO). Superoxide dismutase (SOD, 10 U/ml) prevented 1) the effects of TNF on the hemodynamic responses to U-46619 and ACh and 2) the TNF induced decrease in NO2-. The effects of TNF on lung MPO and effluent O2- were prevented using cyclophosphamide intraperitoneally (100 mg/kg 5 days before, and 50 mg/kg 1 day before, treatment with TNF or control). The data suggest that ROS generated from PMN mediate the decrease in nitric oxide and altered pulmonary vasoreactivity induced by TNF. PMID- 8238534 TI - Respiratory syncytial virus-induced cytokine production by a human bronchial epithelial cell line. AB - Respiratory syncytial virus (RSV) is the most common cause of lower respiratory infection in infants and young children, but the pathogenesis of RSV-induced inflammation is not well defined. We hypothesized that in vitro infection of a human bronchial epithelial cell line (BEAS) would induce production of proinflammatory cytokines. BEAS cells were infected with RSV, and cells and supernatants were assayed for cytokine mRNA and protein changes at several time points after infection. Cytokine mRNA in BEAS cells was measured by polymerase chain reaction of reverse-transcribed RNA from whole cell lysates; cytokine levels in supernatants were measured by bioassay or immunoassay. Our results indicated that interleukin-5ay or immunoassay. Our results indicated that interleukin-8 (IL-8) was induced at 4 h after infection (during the eclipse phase of RSV infection) with accumulation of IL-8 in supernatants by 24 h after infection. Increased levels of IL-6 and granulocyte macrophage colony-stimulating factor in supernatants were only detected by 96 h after infection, during the RSV replicative phase. Interferon-alpha and -gamma transcripts were not detectable at any time point. We conclude that the effects of RSV on airway inflammation may be at least partly mediated by sequential production of proinflammatory cytokines in infected airway epithelium. PMID- 8238535 TI - ATP and UTP increase secretion of bronchial inhibitor by human tracheal gland cells in culture. AB - The effects of ATP and UTP on intracellular Ca2+ levels and on the secretion of the bronchial inhibitor and high-molecular-weight glycoproteins were studied in cultures of human bronchotracheal gland cells. ATP, adenosine 5'-O-(3 thiotriphosphate) (ATP gamma S), and UTP increased intracellular Ca2+ levels in a manner that was partially dependent on the presence of extracellular Ca2+. Other nucleotides (ADP, alpha,beta-methylene ATP, beta,gamma-methylene ATP, and 2 methylthio ATP) and adenosine were ineffective, thus suggesting the presence of a "nucleotide" receptor specific for ATP and UTP. At concentrations similar to those that raised intracellular Ca2+ concentration, ATP, UTP, and ATP gamma S stimulate the secretion of the bronchial inhibitor. ATP and UTP also increase the production of sulfated high-molecular-weight glycoproteins. These results indicate the presence in human tracheal gland cells of a nucleotide receptor that mediates intracellular Ca2+ mobilization and controls the secretion of macromolecules. PMID- 8238536 TI - Release of ED1 fibronectin from matrix of perfused lungs after vascular injury is independent of protein synthesis. AB - Fibronectin (Fn) is an adhesive protein found in the plasma and extracellular tissue matrix. Locally synthesized tissue or cellular Fn (cFn) has extra domains (ED1 and ED2) not present in liver synthesized plasma Fn (pFn). In the lung, Fn is found in the endothelial and epithelial basement membranes, as well as in the interstitial matrix. Utilizing murine monoclonal antibodies to ED1 of cFn, we studied the release of total Fn as well as ED1-Fn into the plasma-free perfusate of the isolated perfused rabbit lung in relation to changes in lung weight due to fluid accumulation after oxidant (H2O2) challenge. Both parameters were also studied after addition of cycloheximide (20 micrograms/ml perfusate) to the perfusion medium to inhibit lung protein synthesis. After continuous H2O2 challenge (11 nmol.ml buffer-1.min-1), there was a 2.25 +/- 0.62 g increase in lung weight over 60 min. Measurement of 125I-labeled albumin clearance at 20 min after the start of H2O2 infusion confirmed an increase in lung endothelial protein permeability after H2O2 treatment. Fn antigen was released into the perfusate as early as 15 min after oxidant challenge. By 60 min, total perfusate Fn increased in H2O2-treated lungs (n = 6) to 2.10 +/- 0.48 micrograms/ml compared with only 0.35 +/- 0.09 micrograms/ml in normal control lungs (n = 5). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of nonreduced samples revealed that the Fn released consisted of primarily intact (440 kDa) Fn as well as Fn fragments. A rapid release of ED1-Fn paralleled the increased release of total Fn.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238537 TI - Elevated lung G protein levels and muscarinic receptor affinity in a mouse model of airway hyperreactivity. AB - A genetic model of airway hyperreactivity has been described in which strains of mice are hyperresponsive (A/J) or hyporesponsive (C3H/HeJ) to intravenous acetylcholine challenge. To determine the mechanism of this differential responsiveness, we compared beta 2-adrenergic and muscarinic cholinergic receptor properties and their coupling to guanine nucleotide binding proteins (G proteins) in peripheral lung membrane fractions from these strains. No significant differences were found between the strains with regard to beta 2-adrenergic or muscarinic receptor density or antagonist affinity. No strain difference was found in beta 2-adrenergic receptor affinity for isoproterenol in the presence or absence of the nonhydrolyzable guanine nucleotide 5'-guanylimidodiphosphate [Gpp(NH)p]. In contrast, affinity of the high-affinity carbachol binding site of muscarinic receptors was threefold greater in A/J lung compared with C3H/HeJ lung (pKH = 7.34 +/- 0.16 vs. 6.79 +/- 0.06, respectively, P < 0.05). In the presence of Gpp(NH)p, this affinity was decreased sevenfold in A/J lung but was not significantly affected in C3H/HeJ lung, suggesting that muscarinic receptors in A/J lung are more effectively coupled to G proteins. Levels of Gs alpha and Gi alpha proteins in peripheral lung were significantly greater in the A/J strain compared with the C3H/HeJ strain (40 and 20% greater, respectively). These studies suggest that airway hyperreactivity in A/J mice is not associated with alterations in beta 2-adrenoceptors, but may be a result of enhanced muscarinic receptor signal transduction due to increased agonist affinity for muscarinic receptors and upregulation of G protein levels. PMID- 8238538 TI - Postinsult treatment with N-acetyl-L-cysteine decreases IL-1-induced neutrophil influx and lung leak in rats. AB - We found that intratracheal administration of interleukin-1 alpha (IL-1) rapidly (5 h) increased leak of 125I-labeled albumin from the blood into the lung (lung leak), influx of neutrophils into lung lavages, lung oxidized glutathione (GSSG) levels, breath hydrogen peroxide (H2O2) concentrations, and lung histological abnormalities in intact rats. Since N-acetyl-L-cysteine (NAC) increases glutathione (GSH) levels in vivo and scavenges oxygen radicals in vitro, we tested the effect of NAC given intravenously on lung changes following intratracheal IL-1 administration. We found that administration of NAC immediately before or 2.5 h after intratracheal administration of IL-1 decreased lung leak, neutrophil influx into lung lavages, and defects in lung histology. NAC treatment also increased blood acid soluble sulfhydryl levels, reduced lung GSSG increases, and decreased breath H2O2 levels in rats given IL-1 intratracheally. The latter findings are consistent with the possibility that NAC is enhancing GSH or other sulfhydryls and, as a result, reducing oxidative stress due to H2O2 or H2O2-derived products. Since postinsult treatment with NAC is effective in this relevant intact animal model of acute lung injury, we speculate that NAC may have promise in the treatment of patients with the adult respiratory distress syndrome. PMID- 8238539 TI - Involvement of neurogenic inflammation in antigen-induced bronchoconstriction in guinea pigs. AB - The role of tachykinins released from sensory nerves in bronchoconstriction induced by antigen was studied in sensitized guinea pigs anesthetized with pentobarbital sodium and pretreated with atropine. The combination of NK2 (SR 48968) and NK1 (CP-96,345) tachykinin-receptor antagonists abolished the increase in total pulmonary resistance (RL) evoked by intravenous capsaicin but did not affect the response evoked by intravenous histamine. A small dose of aerosolized ovalbumin (OVA, 0.1%) produced a small increase in RL that was further increased and markedly prolonged by the neutral endopeptidase (NEP) inhibitor phosphoramidon; this bronchoconstrictor effect of OVA was markedly reduced by the NK2-receptor antagonist and was abolished by the combination of the NK1 and NK2 receptor antagonists together. When a larger dose of OVA (0.5%) was used, a maximal bronchoconstrictor response was obtained. Phosphoramidon did not potentiate this response significantly. The combination of NK1- and NK2-receptor antagonists blunted the response at 5 min only slightly but markedly attenuated the later (10-20 min) response. These results show that tachykinins released from sensory nerves play a significant role in antigen-induced bronchoconstriction in guinea pigs. This effect is exaggerated when the normal modulation of neuropeptides by NEP is inhibited and is mediated predominantly by NK2-receptor activation, with a smaller contribution by NK1 receptors. PMID- 8238540 TI - Effect of gas composition on liquid secretion by explants of distal lung of fetal rat in submersion culture. AB - Throughout gestation, fetal lung is filled with liquid secreted by the pulmonary epithelium. Factors responsible for secretion and for induction of liquid absorption by postnatal lung are poorly understood. We studied effects of "fetal" (3% O2-8% CO2) and "postnatal" (21% O2-5% CO2) gas tensions and of low hormone concentrations [media with 10% charcoal-stripped fetal bovine serum (stFBS) or unstripped FBS] on water content and morphology of distal lung explanted from 14 , 20-, and 22-day fetal (term, 22 days) and 2-day neonatal rats and incubated in submersion culture. Water-to-dry weight ratios of freshly excised fetal whole lung at 20 (6.2) or 22 days (7.0) were greater than that of 2-day postnatal lung (4.7). Culture conditions did not affect water-to-dry weight ratio of cyst forming 14-day explants (10.8) or acystic 2-day postnatal explants (2.4). Fetal gases and stFBS supported cyst formation in [and high water/dry weight (9.2, 12.6) of] 20- and 22-day explants. Cysts also formed in 20-day explants exposed to postnatal gases and stFBS (water/dry weight = 6.5). Other conditions resulted in minimal cyst formation by 20- and 22-day explants and in water/dry weight similar to that of freshly excised and drained distal lung from 22-day fetuses (2.1). Cysts were lined with cuboidal and thin epithelial cells. No cells were ciliated. We conclude that 1) secretion dominates liquid flow across epithelia of fetal rat lung until birth, 2) alveolar epithelium contributes to this secretion, and 3) liquid secretion by fetal distal lung late in gestation is regulated by gas composition. PMID- 8238541 TI - Conservation of structure in ATP-depleted proximal tubules: role of calcium, polyphosphoinositides, and glycine. AB - Increases of intracellular free Ca2+ (Caf) may mediate phospholipid hydrolysis and disintegration in energy-compromised cells; on the other hand, glycine and related amino acids preserve structure. We have examined the effects of increased Caf on phospholipids and structure in ATP-depleted cells, as well as how these actions may be modified by glycine. Incubation of isolated proximal tubules with antimycin A led to ATP depletion, delayed increases of Caf to micromolar levels, polyphosphoinositide (PPI) hydrolysis by phospholipase C, and generalized disintegration of cell structure. Glycine inhibited PPI hydrolysis and preserved cell structure in entirety but did not apparently modify the Caf increases. When overwhelming increases of Caf were induced by the additional presence of a Ca2+ ionophore, glycine did not inhibit either the hydrolysis of PPI or disruption of mitochondria and microvilli. However, the cells remained integrated and unbroken. Incubation in low-Ca2+ medium prevented Caf increases, inhibited PPI hydrolysis, and preserved the structure of mitochondria and microvilli. Nevertheless, there was lethal damage by disintegration of all other membranes. This damage was prevented specifically and completely by glycine. Thus compartments of cells were shown to be differentially susceptible to injury from increased Caf or lack of glycine. Although damage by either factor occurs by distinct mechanisms, glycine also appears to have effects that suppress the deleterious effects of Ca2+ so long as Caf increases are not overwhelming. Our results also suggest that the PPI have a major structural role, which may be compromised by Caf increase during ATP depletion. PMID- 8238542 TI - Luminal and basolateral uptake and receptor binding of IGF-I in rabbit renal proximal tubules. AB - The aim of the present study was to quantify and compare the luminal and basolateral binding and uptake of 125I-labeled insulin-like growth factor I (IGF I) by means of 1) isolated, perfused, proximal tubules combined with electron microscope autoradiography and 2) luminal and basolateral membrane vesicles from rabbit proximal tubules. 125I-IGF-I was added to isolated perfused proximal tubules for 30 min in concentrations of 1.6-3.9 micrograms/l to either the perfusate or the bath. The luminal and basolateral uptake in 30 min averaged 447 and 410 fg/mm, respectively. About 20% of the luminally absorbed IGF-I was digested. Addition of excess unlabeled IGF-I (10(-7) M) to the bath produced complete inhibition of the basolateral binding/uptake, whereas no inhibition of the luminal uptake was seen. Electron microscope autoradiography showed that IGF I after luminal endocytic uptake to a large extent was transported into lysosomes. After basolateral exposure the major portion of the grains was found over the basolateral cell membrane; however, a significant amount was located over endocytic vacuoles and lysosomes in both apical and basal parts of the cells. In both luminal and basolateral membrane vesicles, single-class, high affinity binding sites for IGF-I were found with dissociation constants of 6.3 and 5.7 nM, respectively. Specific binding capacities averaged 2.7 and 25.7 pmol IGF-I/mg protein in luminal and basolateral vesicles. The biochemical data suggest an asymmetric distribution of specific IGF-I receptors in the luminal and basolateral membranes, with a greater abundance of receptors in the latter. The extensive basolateral endocytic binding/uptake of IGF-I compared with that of the luminal in isolated perfused tubules differs considerably from the processing of other peptide hormones. PMID- 8238543 TI - Involvement of PKC-alpha in PDGF-mediated mitogenic signaling in human mesangial cells. AB - Platelet-derived growth factor (PDGF) is a potent mitogen for a variety of cells. The calcium/phospholipid-dependent protein kinase C (PKC) represents a major signal transduction pathway for many growth stimuli including PDGF. Various isoforms of PKC are differentially expressed in the same or in different cells and tissues, and diverse stimuli may selectively activate one or more PKC isoforms. We studied the effect of PDGF on DNA synthesis and on the activity of PKC in human mesangial cells and vascular pericytes in the glomerular microvascular bed. PKC activity was measured as the amount of phosphorylated myelin basic protein-derived peptide substrate in the absence and presence of an inhibitor, a peptide spanning the pseudosubstrate region of PKC. PDGF (15 ng/ml) stimulated PKC activity within 5 min, and the effect was sustained for 60 min. Pretreatment of mesangial cells with 1-(5-isoquinolinesulfonyl)-2 methylpiperazine (H-7), an inhibitor of PKC, abolished the stimulation of PKC and DNA synthesis in response to PDGF. This effect of H-7 was specific, because H-7 did not inhibit the tyrosine phosphorylation of the PDGF receptor in vivo when added to the cells or the in vitro kinase activity in the PDGF beta-receptor immunoprecipitates. Utilizing isotype-specific antibodies against PKC-alpha, beta, or -gamma for immunoprecipitation of PDGF-treated mesangial cell extracts, followed by assay of PKC activity, we demonstrated the activation of PKC-alpha only. Northern blot analysis of mRNA prepared from mesangial cells also revealed two transcripts, 3.7 kb and 1.8 kb, that hybridized with cDNA specific for PKC alpha.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238544 TI - Evidence that separate PGE2 receptors modulate water and sodium transport in rabbit cortical collecting duct. AB - Prostaglandin E2 (PGE2) modulates both water and sodium transport in the rabbit cortical collecting duct (CCD). To determine whether these effects are mediated by separate PGE2 receptors, we compared the effects of PGE2 and its analogue sulprostone in the isolated perfused rabbit CCD. PGE2 increased basal water permeability (hydraulic conductivity), whereas sulprostone did not. PGE2 and sulprostone were equipotent inhibitors of water absorption when it was prestimulated by vasopressin. Pertussis toxin completely reversed the inhibitory effect of sulprostone but only partially reversed the inhibitory effect of PGE2. In contrast, a protein kinase C (PKC) inhibitor, staurosporine, partially reversed the inhibitory effect of PGE2 but had no effect on sulprostone. PGE2 also raised intracellular calcium ([Ca2+]i). This effect is coupled to its capacity to inhibit Na+ absorption. Sulprostone was 10-fold less potent than PGE2 both in raising [Ca2+]i or inhibiting sodium transport. The results suggest sulprostone selectively interacts with a PGE2 receptor coupled to pertussis toxin sensitive inhibition of water permeability. Sulprostone less potently activates a PGE2 receptor coupled to [Ca2+]i, PKC activation, and sodium transport and completely fails to interact with the PGE2 receptor that stimulates water permeability in the collecting duct. These results suggest distinct PGE2 receptors modulate sodium and water transport in the CCD. PMID- 8238545 TI - Interactions of adenosine A1 receptor-mediated renal vasoconstriction with endogenous nitric oxide and ANG II. AB - Renal vasoconstrictor responses to the adenosine A1 agonist N6 cyclopentyladenosine (CPA) were compared in the in situ autoperfused rat kidney to responses evoked by angiotensin II (ANG II), endothelin-1 (ET-1), arginine vasopressin (AVP), carbocyclic thromboxane A2 (CTxA2), phenylephrine (PE), and 5 hydroxytryptamine (5-HT). On the basis of their ED50 values (dose of agonist, in mass units, that produced 50% of maximal response to that agonist), the order of vasoconstrictor potency was ANG II > or = AVP > ET-1 > CPA > 5-HT > or = PE > CTxA2. Dose-response curves to CPA were shallower and maximal responses were weaker than those produced by the other agonists. Maximal responses, the log ED50, and the slope of the dose-response curve to CPA were markedly potentiated in the presence of the nitric oxide (NO) synthase inhibitor N omega-nitro-L arginine methyl ester (L-NAME). Selective antagonism of A1 receptors increased renal blood flow and markedly attenuated CPA-induced renal vasoconstriction in the absence or presence of L-NAME but had no effect on the maximal responses to ANG II. Conversely, AT1 receptor antagonism attenuated renal vasoconstriction produced by ANG II but had little effect on the produced by CPA. These results suggest that endogenous NO modulates renal vasoconstriction produced by A1 receptor stimulation and provide evidence against an interaction between renovascular adenosine A1 and angiotensin AT1 receptors. PMID- 8238546 TI - Modulation of ANG II receptor and its mRNA in normal rat by low-protein feeding. AB - Low-protein feeding results in reduced plasma renin activity (PRA), low prostaglandin production, high intrarenal vascular resistance, and reduced renal plasma flow (RPF) and glomerular filtration rate (GFR) in normal, intact rats. The hemodynamic changes are reversed by converting enzyme inhibitors. In this study, normal rats were fed normal protein (NP) or low protein (LP). PRA was 10.1 +/- 1.3 for NP vs. 5.1 +/- 1.7 ng.ml-1 x h-1 for LP (P < 0.001). Mean arterial pressure fell in both LP and NP during treatment with losartan (DuP-753, a specific angiotensin AT1-receptor inhibitor), but GFR and RPF in LP + losartan became indistinguishable from values obtained in NP and NP + losartan rats. Plasma Na and K and urine excretions of these two electrolytes were unchanged. Angiotensin II (ANG II) binding to isolated glomeruli (n = 19) revealed a dissociation constant of 1.11 +/- 0.22 vs. 1.22 +/- 0.20 nM (not significant) and maximal binding of 763 +/- 89 vs. 432 +/- 75 fmol/mg protein (P < 0.001), indicating an increased number of receptors without changes in affinity in LP. An increased number of receptors in LP compared with NP was also observed by quantitative autoradiography. These results reflect a predominant intrarenal alteration in the response to ANG II in LP. Northern blot and in situ hybridization analysis of the AT1 receptor mRNA showed enhanced gene expression in cortex (glomeruli) and medulla in LP. Dietary protein is an important modulator of the intrarenal actions of ANG II. We show for the first time that protein in the diet modulates the expression of the AT1 receptor gene and that ANG II mediates the hemodynamic changes of LP feeding through the AT1 receptor. PMID- 8238547 TI - Segmentally distinct effects of depolarization on intracellular [Ca2+] in renal arterioles. AB - Experiments were performed to determine the influence of depolarization on intracellular Ca2+ concentration ([Ca2+]i) in renal arterioles and the possible role of voltage-gated Ca2+ channels in these responses. Glomeruli with attached arterioles and thick ascending limb were dissected from rabbit kidney and loaded with fura 2. [Ca2+]i of nonperfused arterioles was monitored using a microscope based dual-excitation wavelength spectrofluorometry system. Afferent arteriolar [Ca2+]i averaged 150 +/- 11 nM (n = 20) when bathed in Ringer solution containing 1.5 mM Ca2+ and 5 mM K+. Replacement of the normal Ringer solution with one containing 100 mM K+ significantly increased afferent arteriolar [Ca2+]i to 196 +/- 12 nM. This response was abolished in the absence of extracellular Ca2+. In the presence of 1 microM nifedipine, 100 mM K+ elicited a 10% decrease in afferent arteriolar [Ca2+]i (P < 0.05). Thus nifedipine reversed the afferent [Ca2+]i response to depolarization, implicating voltage-gated Ca2+ channels as the influx pathway. In contrast to the behavior of afferent arterioles, the 100 mM K+ solution reduced efferent arteriolar [Ca2+]i from 188 +/- 17 to 148 +/- 13 nM (n = 11, P < 0.01), an effect that was not influenced by nifedipine. These observations support a role for voltage-gated Ca2+ channels in eliciting depolarization-induced increases in afferent arteriolar [Ca2+]i while failing to provide evidence for operation of such a mechanism at efferent arteriolar sites. PMID- 8238548 TI - Adrenergic regulation of renin secretion and renal hemodynamics during deliberate hypotension in humans. AB - To assess the relative contributions of neural and nonneural stimuli of renin secretion, the effects of an alpha 1-agonist, phenylephrine (Phe), or a beta agonist, epinephrine (Epi), on plasma renin activity (PRA), renal blood flow (RBF), and glomerular filtration rate (GFR) were compared during sympathetic blockade with epidural hypotensive anesthesia [mean arterial blood pressure (MAP) = 60 and 50 mmHg]. Controls (NaCl) received saline alone to maintain MAP at 50 mmHg. Epi increased PRA (ng.ml-1.h-1) from 0.9 +/- 0.6 to 3.0 +/- 1.5 at 60 mmHg MAP and 4.7 +/- 1.8 at 50 mmHg MAP, with associated decreases in RBF (-33 and 60%, respectively) and GFR (-27 and -53%, respectively). During hypotension with Phe and NaCl, PRA and RBF were unchanged from baseline but GFR decreased. Urinary Na secretion decreased comparably in all three groups. In conclusion, during sympathetic blockade with epidural anesthesia, marked reductions in both renal perfusion pressure and distal nephron Na delivery were insufficient to increase renin secretion. beta-Adrenergic stimulation (e.g., Epi) was required to increase PRA. Epi decreased RBF suggesting an angiotensin II-mediated effect. PMID- 8238549 TI - Na(+)-K(+)-ATPase that redistributes to apical membrane during ATP depletion remains functional. AB - We have previously demonstrated using immunocytochemical, histochemical, and biochemical techniques that ischemia in vivo and ATP depletion in vitro result in dissociation of Na(+)-K(+)-adenosinetriphosphatase (ATPase) from the actin cytoskeleton and redistribution to the apical domain in renal proximal tubule cells. To directly evaluate whether apical Na(+)-K(+)-ATPase retained Na+ pumping activity, a rapidly reversible model of cellular ATP depletion in confluent LLC PK1 cells grown on semipermeable membranes was utilized. Tight-junction integrity, monitored by electrical resistance, was lost during ATP depletion and reestablished during 2 h of ATP repletion. Total cellular Na(+)-K(+)-ATPase activity and total surface membrane [3H]ouabain binding remained constant, but specific apical [3H]ouabain binding increased (7 vs. 26 fmol/filter, P < 0.01). Apical [3H]ouabain binding returned to baseline during 5 h of ATP repletion. Apically applied ouabain was then used to selectively inhibit apical Na(+)-K(+) ATPase. It had no effect on apical-to-basolateral Na+ flux under physiological conditions (1.3 +/- 0.61 vs. 1.27 +/- 0.46 meq.filter-1.30 min-1), but it increased the apical-to-basolateral flux in ATP-depleted and then repleted monolayers (0.39 +/- 0.12 vs. 0.83 +/- 0.27 meq.filter-1.30 min-1, P < 0.01), implying that apical Na(+)-K(+)-ATPase retained Na+ pumping activity. Together, these data imply that ATP depletion induces loss of surface membrane polarity resulting in redistribution of functional proteins to the alternate domain. PMID- 8238550 TI - H(+)-K(+)-ATPase of rat inner medullary collecting duct in primary culture. AB - pH recovery in response to addition of and removal from NH4Cl was examined using 2',7'-bis(2-carboxy-ethyl)-5(6)-carboxyfluorescein fluorescence in primary cultures of inner medullary collecting duct (IMCD) cells from rat kidneys. In 0 K+, pH recovery rate was 0.012 +/- 0.010 U/min; in 5 mM K+, the recovery rate was greater at 0.065 +/- 0.013 U/min (P = 0.026). The H(+)-K(+) adenosinetriphosphatase (H(+)-K(+)-ATPase) inhibitors omeprazole and Sch-28080 and the P-type ATPase inhibitor vanadate significantly inhibited pH recovery at 100, 10, and 5 microM, respectively. The vacuolar H(+)-ATPase inhibitor bafilomycin failed to inhibit pH recovery, but N-ethylmaleimide (NEM) did. A range of Sch-28080 concentrations inhibited ouabain-resistant ATPase activity of microsomes from these cells in a reverse sigmoidal manner, with little inhibition < 1 microM, virtually 100% inhibition > 100 microM, and a 50% inhibitory concentration of approximately 20 microM. Bafilomycin only produced significant inhibition of activity at concentrations well in excess of those that are effective against the vacuolar H(+)-ATPase. The ouabain-resistant ATPase activity in cultured IMCD was also sensitive to vanadate (90% inhibition with 5 microM) but relatively resistant to N,N'-dicyclohexylcarbodiimide and NEM. These results indicate that pH regulation in primary cultures of IMCD cells, presumably reflecting H+ transport, is predominantly due to an H(+)-K(+)-ATPase. PMID- 8238551 TI - 22-Oxacalcitriol: dissection of 1,25(OH)2D3 receptor-mediated and Ca2+ entry stimulating pathways. AB - 22-Oxa-1,25-dihydroxyvitamin D3 (oxacalcitriol, or OCT) is a bioactive analogue of 1 alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] with lower calcemic activity than the parent compound. We investigated the ability of OCT to stimulate 1) genomic pathways mediated by nuclear receptors for 1,25(OH)2D3 versus 2) nongenomic pathways mediated by voltage-sensitive Ca2+ channels in growth phase rat osteosarcoma cells (ROS 17/2.8) and in chick intestine. Effects on nuclear receptor-mediated pathways were evaluated by measuring the ability of OCT to compete with [3H]1,25(OH)2D3 for soluble receptors. We also measured the ability of OCT to increase mRNA encoding osteoblast marker proteins osteopontin (OPN) and osteocalcin (OCN), which are both increased by 1,25(OH)2D3. Effects on Ca2+ entry into osteoblasts were measured using 45Ca2+ influx assays. The rapid stimulation of calcium absorption (transcaltachia) in chick intestine treated with OCT also was measured. We found that OCT bound to the nuclear receptor with lower binding affinity [relative competitive index (RCI) = 48.1 for ROS 17/2.8; RCI = 14.8 for chick intestine] than 1,25(OH)2D3 (RCI = 100). Like 1,25(OH)2D3, OCT increased mRNA levels of OPN and OCN in ROS 17/2.8 cells over a 48-h period. In contrast, OCT had no effect on transmembrane influx of 45Ca2+ across ROS cell membranes, whereas uptake was stimulated within 1 min by 1 nM 1,25(OH)2D3. In transcaltachia assays in perfused duodenum, OCT stimulated absorption with a maximum response at 6.5 nM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238552 TI - Peritubular endothelial cell proliferation in mice during compensatory renal growth after unilateral nephrectomy. AB - The proliferation of peritubular endothelial cells during compensatory renal growth (CRG) following unilateral nephrectomy in mice was investigated using a labeling index. The labeling index of peritubular endothelial cells increased 6 h after uninephrectomy and decreased to the normal level within 72 h. Immunohistochemical study revealed that c-myc protein was expressed in the nuclei of both cortical tubular cells and peritubular endothelial cells. Furthermore, intravenous injection of anti-basic fibroblast growth factor (bFGF) neutralizing antibody just after uninephrectomy led to significant inhibition of proliferation of peritubular endothelial cells, but not tubular cells. These results indicate that peritubular endothelial cells proliferate transiently during CRG and that bFGF plays an important role for the growth regulation of that cell in the kidney. PMID- 8238553 TI - Dynamic, short-term coupling between changes in arterial pressure and urine flow. AB - Pressure diuresis refers to the direct effect of arterial pressure (AP) on the rate of urine flow (UF). On the basis of computer modeling, pressure diuresis has been viewed as a long-term mechanism that acts to set the level of the blood volume and, thus, the steady-state AP. There are no systematic studies, however, on the rapidity with which changes in AP induce changes in UF in vivo. Therefore, we measured the delay between induced changes in AP and the subsequent change in UF. Nine anesthetized rats were instrumented with arterial, venous, and ureteral catheters. AP and UF were measured every 2 s, while acute changes in AP were induced by 1) occlusion of the aorta above or below the renal vessels; 2) brief tail pinch; or 3) intravenous administration of acetylcholine (1 microgram), phenylephrine (1 microgram), or angiotensin II (0.1 microgram). The rapidity of the urinary response to induced changes in AP was determined by calculating the delay between a significant change in AP (+/- 2 SD from baseline) and a significant change in UF. The delay averaged 6.0 +/- 0.5 s for all conditions. Also, examining the relationship between the magnitude of the induced changes in AP and the magnitude of the responses in UF revealed an exponential influence of AP on UF. That is, there were proportionately larger changes in UF compared with AP (< or = 10 times greater magnitude) in response to the experimental interventions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238554 TI - Role of apical and basolateral secretion in turnover of glutathione in LLC-PK1 cells. AB - Previous clearance measurements have established that the rapid turnover of renal proximal tubular glutathione is in part due to apical secretion and degradation by gamma-glutamyltranspeptidase, an ectoenzyme that is primarily associated with the brush-border membrane. The relationship between glutathione turnover and secretion was further characterized using confluent cultures of LLC-PK1 cells grown on nitrocellulose supports. The resulting cell layer was impermeable to [3H]inulin and exhibited a polarized expression of gamma-glutamyltranspeptidase. Incubating cells with 5 mM buthionine sulfoximine, an inhibitor of glutathione synthesis, produced an 86% inhibition of [35S]cystine incorporation into intracellular glutathione. Under these conditions, the prominent intracellular pool of glutathione turns over with an apparent half-life of 4 h and a first order rate constant of 0.17 h-1. This turnover is unaffected by pretreatment with AT-125, an inhibitor of gamma-glutamyltranspeptidase. The rate of accumulation of glutathione in the apical and basolateral medium of cells pretreated with AT-125 was 22 and 34 nmol.mg protein-1.h-1, respectively. The combined secretion was equivalent to the calculated turnover rate of intracellular glutathione (57 nmol.mg protein-1.h-1). Therefore, the combined processes of apical and basolateral secretion can account for the turnover of intracellular glutathione in LLC-PK1 cells. PMID- 8238555 TI - Effects of in vivo and in vitro alkali treatment on intracellular pH regulation of OMCDis cells. AB - To examine functional changes of the transporters in the inner stripe of the outer medullary collecting ducts (OMCDis) by the peritubular acid-base status, in vitro microperfusion using the acetoxymethyl ester of 2',7'-bis(2-carboxyethyl) 5(6)-carboxyfluorescein was performed. Cell alkalinization systems were assessed by the recovery rate (dpHi/dt) of intracellular pH (pHi) after intracellular acid loading by NH(4+)-NH3 prepulse with bath amiloride. In alkali-loaded rabbits (0.15 M NaHCO3 drinking for 14 days), dpHi/dt showed a significant decrease (1.80 +/- 0.29 pH units/s x 10(3)) compared with either control (3.30 +/- 0.59) or acid loaded rabbits (0.15 M NH4Cl drinking for 14 days, 3.05 +/- 0.46). The difference of dpHi/dt between control and alkali-loaded rabbits was eliminated by lumen N ethylmaleimide (NEM), suggesting that H+ pump activity was decreased. The effect of in vitro alkali treatment (50 mM HCO3-, pH 7.7) for 3-4 h was also examined. This incubation significantly decreased the dpHi/dt (1.83 +/- 0.35) compared with the time control experiments (3.18 +/- 0.28), whereas no significant difference was seen in the presence of lumen NEM. Anion exchanger activity, as determined from the pHi changes after Cl- addition to the bath, showed no significant change with in vivo or in vitro alkali treatment. The results indicate that cell function of the OMCDis is regulated in response to the peritubular acid-base environment via changes in the H(+)-adenosinetriphosphatase. PMID- 8238556 TI - NHE3: a Na+/H+ exchanger isoform of renal brush border. AB - Na+/H+ exchangers in the brush-border (luminal, apical) membrane of renal proximal tubules are responsible for active, transcellular reabsorption of NaHCO3 and NaCl. Although well characterized kinetically, the protein that mediates Na+/H+ exchange in the renal brush border has not been identified. Several Na+/H+ exchanger genes, including NHE1, NHE2, NHE3, and NHE4, are expressed in the kidney. To identify the NHE3 gene product and to determine its cellular and subcellular localization in the rabbit kidney, an NHE3-isoform-specific antibody was prepared. Guinea pigs were immunized with purified fusion protein containing the carboxy-terminal 40 amino acids of NHE3 (fpNHE3-C40). After affinity purification, immune sera demonstrated specific reactivity to the NHE3 sequence within the fusion protein as well as to an 80-kDa polypeptide expressed in NHE3 transfected LAP1 cells. Western blot analysis showed that anti-fpNHE3-C40 specifically reacted with an 80-kDa protein that is relatively enriched in renal brush-border membrane compared with basolateral membrane. Immunocytochemical studies confirmed that the Na+/H+ exchanger isoform NHE3 is expressed along the microvillar membrane of the brush border of proximal tubule cells in the rabbit kidney. PMID- 8238557 TI - Inhibition of adenosine-mediated coronary vasodilation exacerbates myocardial ischemia during exercise. AB - Persisting coronary vasoconstrictor tone that is responsive to exogenous adenosine administration has been demonstrated during myocardial ischemia. Therefore, the role and extent of endogenous adenosine-mediated coronary vasodilation in opposing coronary vasoconstriction within regions of ischemic myocardium was investigated in 10 chronically instrumented exercising dogs. Studies were performed on dogs with left circumflex coronary artery stenosis during treadmill exercise (6.5 km/h, 6% grade), while myocardial blood flow was measured with radioactive microspheres. Blood flow was measured before and again after inhibition of the effects of endogenously produced adenosine through combined inactivation of adenosine and adenosine receptor antagonism by the administration of intracoronary adenosine deaminase (ADA) (5 micrograms.kg-1 x min-1 x 10 min) plus 8-phenyltheophylline (8-PT) (5 mg/kg i.v.), respectively. Coronary perfusion pressure was held equal during both conditions at approximately 41 mmHg with a hydraulic occluder. During exercise in the presence of a coronary stenosis, blood flow was reduced in all layers of myocardium in regions supplied by the stenosed left circumflex coronary artery compared with blood flow in regions of myocardium supplied by the nonstenotic left anterior descending coronary artery. After ADA plus 8-PT, myocardial blood flow (in ml.min 1 x g-1) was further reduced in all layers of myocardium in regions supplied by the stenotic left circumflex coronary artery compared with baseline (subendocardial layer 0.44 +/- 0.09 vs. 0.67 +/- 0.13 ml.min-1 x g-1, mean transmural flow 0.92 +/- 0.13 vs. 1.25 +/- 0.2 ml.min-1 x g-1, both P < 0.05). Blood flow in regions of myocardium supplied by the nonstenotic left anterior descending coronary artery were unchanged following ADA plus 8-PT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238558 TI - Lidocaine attenuates both mechanical and metabolic changes induced by hydrogen peroxide in the rat heart. AB - The effect of lidocaine on the hydrogen peroxide (H2O2)-induced mechanical and metabolic changes was studied in the Langendorff rat heart, which was perfused aerobically at a constant flow rate and paced electrically. H2O2 (600 microM) increased the left ventricular end-diastolic pressure (i.e., mechanical dysfunction), decreased the tissue levels of ATP and creatine phosphate (i.e., metabolic change), and increased the tissue level of malondialdehyde (MDA). The mechanical and metabolic alterations induced by H2O2 were attenuated by lidocaine (50 or 200 microM). Lidocaine also attenuated the H2O2-induced increase in MDA. Nevertheless, lidocaine modified neither the myocardial ferrylmyoglobin level that was increased by H2O2 nor the H2O2 concentration in vitro and did not affect the mechanical function and energy metabolism of the normal heart. These results suggest that lidocaine attenuates both mechanical and metabolic changes induced by exogenous H2O2. The protective action of lidocaine on the H2O2-induced changes is due to neither energy-sparing nor H2O2-scavenging effect, but probably due to its effect to reduce the H2O2-induced lipid peroxidation. PMID- 8238559 TI - Coronary artery acidosis: pH and calcium pump stability. AB - To study the effects of prolonged exposure to different pH, pig coronary artery smooth muscle subcellular fraction F3 enriched in sarcoplasmic reticulum was preincubated at 0 or 37 degrees C and pH 6.4-7.8 and then used for monitoring the rate of the oxalate-stimulated component of the ATP-dependent azide-insensitive Ca2+ uptake at constant pH (6.8) and temperature (37 degrees C). Ca2+ uptake by F3 decreased with the increasing preincubation pH. The loss of Ca2+ uptake was more rapid upon preincubation at 37 degrees C than at 0 degrees C. Dithiothreitol (DTT), when included in the preincubation solution, protected against the loss. Sucrose, KCl, ATP, and ATP plus CaCl2 protected slightly, and glutathione, catalase, superoxide dismutase, azide, ascorbate, CaCl2, ethylene glycol-bis(beta aminoethyl ether)-N,N,N',N'-tetraacetic acid, mannitol, mercaptopropionylglycine, and FeSO4 had only marginal or no effects. Efflux of accumulated Ca2+ was more rapid from membranes that had been preincubated at 37 degrees C than from those preincubated at 0 degrees C, but it was not affected by the preincubation pH or by DTT. The loss of Ca2+ uptake due to incubation at pH 7.8 accompanied a decrease in the 115-kDa Ca(2+)-dependent acylphosphate formation due to the Ca2+ pump. The presence of DTT in the preincubation mixture increased the acylphosphate level in the control and protected against its loss at the preincubation pH 6.8 or 7.8. Thus, in the membranes isolated from coronary artery, acidosis may protect against damage to the sarcoplasmic reticulum Ca2+ pump by protonation of a key sulfhydryl group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238560 TI - Effects of 2,3-butanedione monoxime on sarcoplasmic reticulum of saponin-treated rat cardiac muscle. AB - We have studied the effects of 2,3-butanedione monoxime (BDM) on the sarcoplasmic reticulum (SR) of saponin-treated rat cardiac trabeculae. Rapid application of 20 mM caffeine released Ca2+ from the SR, which was detected using the fluorescent Ca2+ indicator indo 1. The amplitude of the caffeine-induced Ca2+ transient was used as an index of the Ca2+ content of the SR before, during, and after exposure to various concentrations of BDM. BDM (1-5 mM) had little effect on caffeine induced Ca2+ release. At these levels of BDM, force was inhibited predominantly by a direct action of BDM on the myofilaments. However, with higher concentrations (5-30 mM), BDM caused a concentration-dependent decrease in the amount of Ca2+ released from the SR in response to caffeine. This action of BDM may contribute to the negative inotropic effect of the drug in intact cardiac preparations by reducing the amount of Ca2+ available for release during systole. Rapid application of BDM induced a net release of Ca2+ from the SR. Both BDM and caffeine-induced Ca2+ releases were abolished following treatment of the muscle with 10 microM ryanodine. BDM failed to release Ca2+ in the absence of ATP or after substitution of ATP with nonhydrolyzable adenine nucleotides. In contrast, caffeine released Ca2+ in the absence of ATP. The possible involvement of the Ca(2+)-uptake pump in the action of BDM on the SR is discussed. PMID- 8238561 TI - Characterization of alpha-adrenoceptor gene expression in arterial and venous smooth muscle. AB - Six genes coding for three unique alpha 1- (1A, 1B, 1C) and three unique alpha 2- (2A, 2B, 2C) adrenergic receptor (AR) subtypes have been cloned. Ligand binding and contractile studies have demonstrated that both alpha 1- and alpha 2-ARs can exist on vascular smooth muscle (VSM) cells, although less is known about the relative distribution and specific subtypes in different vascular segments. In the present study polymerase chain reaction (PCR) analysis was used to characterize the species of alpha-AR messenger RNA (mRNA) present in freshly isolated rat thoracic aortic media and vena cava and in cultured VSM cells (passage 2) derived from both sources. To prevent possible contamination of VSM mRNA, aortic media was separated from adventitia, and vessels were denuded of endothelial cells. Oligonucleotide primers specific for each of the six adrenergic genes were synthesized and used to probe for the presence of alpha-AR mRNA species after reverse transcription of total cellular RNA to cDNA. PCR amplified AR transcripts were distinguished by the size of amplified DNA fragments and unique restriction endonuclease cleavage. Expression of alpha 1C- or alpha 2C-mRNA was not detected in vascular tissues or cultured VSM cells, although the alpha 2C-primers detected the expected alpha 2C expression in cerebral cortex. Only alpha 1A-mRNA was detected in aortic adventitia. VSM from aorta expressed alpha 1A-, alpha 1B-, and alpha 2A-mRNA, and this pattern was preserved in cultured aortic VSM. Vena cava also expressed both alpha 1A and alpha 1B; however only alpha 2B-mRNA was detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238562 TI - Distribution of microvascular pressure in arteriolar vessel trees of ventilated rabbit lungs. AB - We have developed a new in vivo microscopic technique for comprehensive measurements of vessel diameter, segment length, and red blood cell velocity in discrete arteriolar vessel trees of the lung. In anesthetized and mechanically ventilated rabbits, a transparent window was implanted into the right thoracic wall. We injected fluorescently labeled red cells to visualize blood flow and to measure red blood cell velocity. The distribution of microvascular pressures was simulated in a computer model based on morphometric and microhemodynamic data. Of the total pulmonary vascular pressure drop from pulmonary artery to left atrium, on average 2.5% occurred in distal arteriolar vessel trees with main trunk diameters of 73-111 microns. Along the pathlength from main trunk to terminal arterioles (0.18-2.79 mm), the pressure drop ranged between 0.06 and 0.94 mmHg. The pressure drop along individual pathways correlated significantly with pathlength of terminal arterioles, whereas red blood cell velocity did not. The results indicate that in terminal arteriolar vessel trees of the ventilated rabbit lung the resistance to blood flow is low, and the heterogeneity of microvascular pressures in arterioles feeding capillary networks is high. PMID- 8238563 TI - Loss of hypoxic pulmonary vasoconstriction in chronic pneumonia is not mediated by nitric oxide. AB - In pulmonary inflammatory processes such as pneumonia there is diminished hypoxic pulmonary vasoconstriction (HPV). We investigated whether the attenuated HPV in pneumonia is a due to excess nitric oxide (NO) release. Sprague-Dawley rats were anesthetized, and a slurry (0.06 ml) of infected agar beads (containing 6 x 10(5) Pseudomonas aeruginosa organisms) or control (sterile) beads was then injected into a distal bronchus through a tracheotomy. After the establishment of a chronic P. aeruginosa pneumonia (7-10 days later) animals were instrumented for hemodynamic monitoring, and the response to exposure to hypoxic gas (fraction of inspired O2 = 0.08) was recorded before and after the administration of NG monomethyl-L-arginine (L-NMMA; 50 mg/kg), an inhibitor of NO synthesis. The hypoxic pressor response, as assessed by the absolute increase in pulmonary arterial pressure (PAP) and total pulmonary resistance (TPR), was reduced in infected animals compared with control animals. The change in PAP and TPR was 8.5 +/- 0.7 and 0.053 +/- 0.007, respectively, in control animals compared with 5.9 +/- 0.5 and 0.041 +/- 0.011 in infected animals. After L-NMMA the increase in PAP and TPR during hypoxia was greater in both control and infected animals. However, treatment with L-NMMA did not affect the difference between control and infected animals. We conclude that excess release of NO does not account for the attenuated hypoxic pressor response in pneumonia. PMID- 8238564 TI - Intracellular calcium chelator, BAPTA-AM, prevents cocaine-induced ventricular fibrillation. AB - Cocaine is a potent cardiac stimulant that can provoke lethal cardiac events, including ventricular fibrillation (VF). The cocaine-induced accumulation of intracellular calcium could contribute significantly to the development of these lethal arrhythmias. To test this hypothesis, VF was induced in 12 mongrel dogs by the combination of cocaine (1.0 mg/kg) and a 2-min coronary occlusion during exercise. This test without cocaine failed to induce arrhythmias. Pretreatment with the intracellular calcium-specific chelator 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA-AM; 1.0 mg/kg iv) prevented VF in 8 of 12 animals (P < 0.001) and delayed the onset of lethal arrhythmias in 3 of the remaining animals. Cocaine induced significant increases in left ventricular (LV) systolic pressure (control 154.7 +/- 8.7, cocaine 167.4 +/- 8.4 mmHg), heart rate (control 195.9 +/- 6.1, cocaine 222.3 +/- 10.6 beats/min), and LV maximum rate of pressure development (dP/dtmax; control 5,251 +/- 317.6, cocaine 6,016 +/- 435.1 mmHg/s). BAPTA-AM attenuated the increase in LV dP/dtmax (BAPTA-AM 4,591 +/- 479.3 mmHg/s) and LV systolic pressure (BAPTA-AM 154.5 +/- 6.8 mmHg). Because vascular muscle relaxation could contribute to the cardioprotection, the cocaine and exercise plus ischemia test was repeated after nitroprusside. The nitroprusside prevented cocaine-induced increases in LV systolic pressure but failed to prevent VF. These data suggest that BAPTA-AM may prevent cocaine-induced VF independently of its vascular actions. PMID- 8238565 TI - Interaction of hypokalemia and ventricular dilatation in isolated rabbit hearts. AB - Potassium depletion has been implicated in the genesis of ventricular arrhythmias, especially in patients with congestive heart failure, but the nature of this interaction is unknown. We studied the electrophysiological effects of varying concentrations of extracellular K+ in 24 isolated, retrogradely perfused rabbit hearts. The left ventricular cavity was dilated with a fluid-filled balloon. Left ventricular effective refractory period decreased significantly (from 119 +/- 3 to 105 +/- 3 ms) with ventricular dilatation but was not influenced by K+ concentration. Conduction velocity was not changed by dilatation but was decreased by hypokalemia (81 +/- 2 cm/s, K+ = 4.9 mM; 75 +/- 2 cm/s, K+ = 3.5 mM; 70 +/- 2 cm/s, K+ = 2.5 mM; P < 0.05). No ventricular arrhythmias were induced in undilated hearts at a K+ concentration of 4.9 mM. Induced ventricular fibrillation was more frequent (38%; P < 0.01) in the dilated heart and increased markedly (92%; P < 0.01) with coexisting hypokalemia. The incidence of induced fibrillation was highly correlated with the wavelength of the ventricular impulse (wavelength = refractory period x conduction velocity). Thus dilatation and hypokalemia have very different electrophysiological effects that can interact synergistically to predispose the heart to reentrant arrhythmias in this model. PMID- 8238566 TI - Renin-angiotensin system and minoxidil-induced cardiac hypertrophy in rats. AB - Besides cardiac volume overload, cardiac sympathetic activity and the renin angiotensin system (RAS) are activated by arterial vasodilators such as minoxidil. To evaluate the possible involvement of the RAS in the development of minoxidil-induced cardiac hypertrophy, we assessed in normotensive rats minoxidil induced changes in cardiac and plasma renin activity (PRA) and the potential of chronic treatment with the angiotensin-converting enzyme (ACE) inhibitor enalapril and the nonpeptide angiotensin II receptor blocker losartan to prevent minoxidil-induced cardiac hypertrophy. PRA increased in parallel with the increase in cardiac filling pressures and development of cardiac hypertrophy, whereas the increase in cardiac renin activity was delayed as compared with these changes. Losartan did not decrease left ventricular end-diastolic pressure (LVEDP) but prevented the remodeling of the heart by minoxidil. In contrast, enalapril nearly normalized LVEDP but did not affect the hypertrophic response of the heart. The losartan data indicate that the RAS is involved in the minoxidil induced cardiac hypertrophy either directly (e.g., by mediating the hypertrophic response of the heart to cardiac volume overload) or indirectly (e.g., by potentiating cardiac sympathetic activity). The ineffectiveness of enalapril has no obvious explanation but may possibly indicate ineffective blockade of angiotensin II formation in the heart in this model. PMID- 8238567 TI - Effect of hypoxemia on the cardiovascular response to intracranial hypertension in postnatal lambs. AB - Large increases in intracranial pressure in fetal sheep result in more potent peripheral vasoconstriction and better maintenance of cerebral O2 consumption (CMRO2) than in postnatal sheep. The fetus is exposed to a lower PO2. We tested the hypothesis that low PO2 in postnatal lambs potentiates peripheral vasoconstriction and better maintains cerebral perfusion pressure and CMRO2. Pentobarbital-anesthetized lambs, 2-7 days old, were ventilated with either room air (n = 7) or a low O2 mixture to reduce arterial O2 saturation to 50% (n = 7). Elevation of intracranial pressure to within 3-5 mmHg of baseline mean arterial pressure for 30 min by ventricular fluid infusion initially caused a similar increase in arterial pressure in the normoxic [11 +/- 3 (SE) mmHg] and hypoxic (14 +/- 2 mmHg) groups. Plasma catecholamines increased more rapidly in the hypoxic group. However, plasma vasopressin levels were substantially elevated by hypoxia alone and failed to increase further with elevated intracranial pressure. Moreover, there was no significant difference between groups in the steady-state increase in arterial pressure, and microsphere-determined blood flow to intestines, kidney, skin, and muscle did not decrease in either group. Consequently, cerebral perfusion pressure, regional cerebral blood flow, and CMRO2 were reduced similarly in both groups. Therefore, hypoxemia failed to potentiate the postnatal pressor response. Low PO2 is unlikely to be the major mechanism for the potent Cushing response in the fetus. PMID- 8238568 TI - Daily spontaneous running did not alter vagal afferent reactivity. AB - Exercise training alters the cardiopulmonary baroreflex regulation of the circulation; however, the mechanisms responsible are unknown. One possibility is an enhanced afferent response to cardiopulmonary stimulation. We therefore tested the hypothesis that daily spontaneous running (DSR) would enhance cardiopulmonary vagal afferent responses to mechanical (increase in left atrial pressure, LAP) and chemical (phenyl biguanide, PBG) stimulation. Reactivity of single-fiber cardiopulmonary vagal afferents was evaluated in 16 control and 12 DSR anesthetized Sprague-Dawley rats. Rats were weaned at 3 wk of age and randomly assigned to a control or DSR group. Eight to twelve weeks of DSR was associated with a 27% increase in heart weight-to-body weight ratio (3.27 +/- 0.08 vs. 2.56 +/- 0.05 g/kg, P < 0.001) and resting bradycardia (394 +/- 10 vs. 421 +/- 8 beats/min, P = 0.036). However, DSR did not alter the stimulus-response curves to increases in LAP (frequency of discharge vs. LAP) for either the high-frequency (maximum response, sedentary 59.6 +/- 3.2, DSR 60.1 +/- 5.0 spikes/s) or low frequency (maximum response, sedentary 20.0 +/- 2.9 DSR 20.6 +/- 3.9 spikes/s) receptors. Dose-response curves to chemical stimulation (spikes/s vs. PBG dose) were also not altered by DSR. Thus DSR did not change vagal afferent reactivity to mechanical or chemical stimulation. PMID- 8238569 TI - Pyruvate enhances recovery of rat hearts after ischemia and reperfusion by preventing free radical generation. AB - Pyruvate protects myocardium from ischemic and anoxic injury, effects that have been attributed to beneficial metabolic alterations. Pyruvate also reacts with hydrogen peroxide in vitro, and pyruvate prevents free radical injury in other organs. Hearts supplied with 2 mM of pyruvate with glucose during reperfusion recovered significantly more mechanical function (81%) than those provided with 2 mM of acetate (which does not react with free radicals) and glucose (49%) or glucose alone (27%). Pyruvate significantly reduced free radical generation during reperfusion as measured with electron spin resonance using the spin-trap 5,5-dimethyl-1-pyrroline-1-oxide. In a model of direct oxidant stress, hearts were perfused with 0.28 mM of hydrogen peroxide. In this model, loss of function was almost entirely prevented by addition of 2 mM of pyruvate. From these results we conclude an important mechanism of protection when pyruvate is supplied during reperfusion is limitation of oxygen-derived free radical damage. PMID- 8238570 TI - Two-port analysis of systemic venous and arterial impedances. AB - Hemodynamic properties of the systemic vasculature were measured in eight anesthetized dogs using two-port impedance analysis. Blood pressures and flows were measured at the aortic root and the caval-atrial junction. Impedances were computed from 0.05 to 20 Hz to characterize the systemic vasculature. Pseudorandom variations in flow were produced with an extracorporeal perfusion system. Impedance measurements were made at carotid baroreceptor pressures of 50, 125, and 200 mmHg. A six-parameter lumped-element model best fitted the measured impedance spectra. At 125 mmHg, the mean parameter values were venous inertance, 13.5 g.kg.cm-4; venous and arterial compliances, 0.769 and 0.0214 ml.mmHg-1.kg-1; venous and arterial characteristic impedances, 0.028 and 0.084 mmHg.kg.min.ml-1; and arterial-to-venous small-vessel resistance, 0.706 mmHg.kg.min.ml-1. Regression analysis showed significant dependence of small-vessel resistance on baroreceptor pressure. The other parameters were not dependent on carotid sinus pressure, which is consistent with baroreflex control of venous unstressed volume but not compliance. We conclude that two-port impedance analysis is a useful tool for studying venous hemodynamics and the dynamic coupling between the veins and the right heart. PMID- 8238571 TI - Impulse propagation in the Purkinje system and myocardium of intact dogs. AB - To characterize Purkinje activation (PA) patterns, we studied 15 alpha-chloralose anesthetized dogs. During left ventricular basal or apical pacing, PA was mapped with a row of four to eight multipolar electrodes placed in the left ventricular wall to record Purkinje and muscle potentials. PA times increased linearly with distance from the pacing site (r = 0.8, slope 1.8 +/- 0.15 mm/ms, P < 0.05). With a single premature stimulus (S2) delivered from the apical or basal site, delay of PA times was first evident at remote sites (> or = 50 mm) with S1-S2 = 185 +/- 5 (SD) ms, while at peripacing sites (< or = 10 mm), delay was first evident when S1-S2 = 176 +/- 6 ms (Purkinje relative refractory period; P < 0.05). With S2,...S5 such that S1-S2 = S2-S3, and so on, an alternating mode of conduction, both temporally and spatially, occurred. The alternation was due to refractoriness and conduction interactions. With short premature intervals, remote coupling intervals were 30 ms less than the premature intervals. Conduction delay in Purkinje fibers with premature stimuli allowed remote epicardium to be activated by alternate routes. Conduction of constant-rate stimuli in the Purkinje system is relatively uniform. With single premature stimuli, conduction is delayed first remotely from the pacing site, whether apical or basal stimulation was employed. When S1-S2 is less than Purkinje relative refractory period, the usual endocardial-to-epicardial activation sequence is altered. Multiple premature stimuli cause remote coupling intervals to alternate. PMID- 8238572 TI - Myocardial metabolism in chronic reperfusion after nontransmural infarction in pig hearts. AB - The purpose of these studies was to evaluate metabolic behavior in a 4-day reperfusion model in pigs after induction of subendocardial infarction. Two groups of swine [sham and intervention (Int) groups, n = 7) and 10 hearts per group, respectively] were prepared comparably with two surgical procedures separated over 4 days. In the Int group at the time of the first surgery, coronary flow in the left anterior descending (LAD) circulation was partially restricted (by 60%) for 60 min and was then reperfused. LAD myocardium at the time of the second surgery in both groups was extracorporeally perfused aerobically (5.9 +/- 0.2 ml.min-1.g dry wt-1) for 60 min and infused by equilibrium labeling with [U-14C]-palmitate and [5-3H]glucose to estimate fatty acid oxidation and exogenous glucose utilization. During extracorporeal perfusion, regional myocardial shortening and oxygen consumption were comparable between groups despite a marginal impairment in ATP resynthesis by mitochondria (26% decrease, P < 0.071) in Int hearts and a significant decline in mitochondrial respiration (45% decrease in respiratory control rate, P < 0.008; and 41% decrease in state 3 respiration, P < 0.032) as compared with sham hearts. Fatty acid oxidation described by 14CO2 production was 34.00 +/- 4.72 mumol.h-1.g dry wt-1 (averaged from 30-60 min of perfusion) in sham hearts but was decreased (by 48%, P < 0.004) in Int hearts. This reduction in fatty acid utilization may in part be explained by declines in the observed activity of the mitochondrial membrane transporter enzyme, carnitine palmitoyltransferase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238573 TI - Resetting of regional preload due to ventricular shape change alters diastolic and systolic performance. AB - The diastolic and systolic pressure of one ventricle is increased by an increase in volume and/or pressure of the opposite ventricle; however, a mechanism for the ventricular interaction remains unclear. We hypothesized that the shape change of one ventricle elicited by the opposite ventricle would lead to resetting of the regional length, which may explain the ventricular interaction. We used 15 cross circulated isovolumically contracting canine hearts in which both ventricular volumes were independently controlled. Diastolic regional segment area was calculated by multiplying circumferential and longitudinal lengths on right ventricular free wall (RVFW; n = 6), interventricular septum (IVS; n = 11), and left ventricular (LV) FW (n = 12). The regional area at relatively small volumes of both ventricles were expressed as 100%. With constant RV volume, increasing LV from 7 to 19 ml increased RV diastolic and systolic pressures by 2.7 and 5.5 mmHg, respectively. Conversely, increasing RV volume increased LV diastolic and systolic pressures by 2.3 and 7.5 mmHg, respectively. Increasing LV volume increased RVFW regional area from 121.0 to 124.6% (P < 0.01) and increased IVS regional area from 103.3 to 108.7% (P < 0.01), whereas the RV volume was held constant. Increasing RV volume also increased LVFW and IVS regional areas from 109.9 to 111.6% (P < 0.01) and from 106.8 to 108.9% (P < 0.05), respectively. Ventricular shape change elicited by ventricular interaction will increase the regional wall area, even though the volume of the chamber is unchanged. The increase in the regional area alters the position of the tissue on its resting and active length-tension relations and, thus, leads to enhancement of the chamber pressure. PMID- 8238574 TI - Effects of hypothyroidism and hypertension on myocardial perfusion and vascularity in rabbits. AB - We considered the role of thyroid hormones in the growth of the coronary microvasculature during various levels of afterload. Arterial pressure was increased for 3 mo in hypothyroid and euthyroid rabbits via the Page (1-kidney, 1 wrap) method. Systolic pressures (mmHg +/- SE) indicated that the groups could be characterized as follows: euthyroid sham normotensive (103 +/- 5), euthyroid Page hypertensive (158 +/- 24), hypothyroid Page normotensive (110 +/- 5), and hypothyroid sham hypotensive (87 +/- 5). The hypothyroid groups were characterized by bradycardia. Left ventricular weight-to-body weight ratios were higher in the Page groups than in the controls. Minimal coronary vascular resistance (MCVR) was elevated in the euthyroid Page group but was not affected by hypothyroidism. Lumen diameters of the major resistance vessels tended to be larger in the hypertensive rabbits. Capillary length density was greater in the hypothyroid than in the euthyroid groups. We conclude that 1) the elevated MCVR in the hypertensive rabbits is due mainly to a failure of the major resistance vessels to increase in size or number rather than to a decrease in vascular lumen diameter, and 2) hypothyroidism does not affect maximal myocardial perfusion or lumen diameter of resistance vessels but facilitates capillary growth. The latter may be due to the presence of bradycardia. PMID- 8238575 TI - Effects of isoproterenol on myocardial relaxation rate: influence of the level of load. AB - The present work was undertaken to test the hypothesis that the level of the load faced by the myocardium influences the effects of isoproterenol on relaxation rate. Responses to cumulative doses of isoproterenol (from 10(-10) to 10(-6) M) were studied in rat left ventricular papillary muscle stimulated 12 beats/min at 29 degrees C in 0.5 mM extracellular calcium and preloaded at initial muscle length corresponding to apex of length-active tension curve (Lmax; group 1, n = 20) or at 95% of Lmax (group 2, n = 9). A control group (group 3, n = 8) was studied every 15 min for 75 min. We measured maximum unloaded shortening velocity (Vmax), normalized positive and negative peak force derivatives (+dF and -dF, respectively) of the fully isometric twitch, and peak lengthening velocity of the isotonic twitch with preload only (Vlmax). In group 1, Vmax and +dF increased under 10(-10) and 10(-9) M isoproterenol, respectively, and -dF increased under 10(-9) M isoproterenol (115 +/- 13 vs. 96 +/- 12 mN.s-1.mm-2, P = 0.01). Conversely, Vlmax increased under 10(-7) M isoproterenol only (2.34 +/- 0.19 vs. 1.45 +/- 0.18 Lmax/s, P < 0.001). In group 2, both -dF and Vlmax increased under 10(-7) M isoproterenol only (P = 0.015 and 0.011, respectively). In group 3, -dF and Vlmax did not vary in time. Our results suggest a load-revealed (or length revealed) difference in the dose dependence of the various biochemical processes involved in the effects of isoproterenol during myocardial relaxation. PMID- 8238576 TI - Arrhythmogenic effects of catecholamines are decreased in heart failure induced by rapid pacing in dogs. AB - Increased circulating catecholamines are considered to be arrhythmogenic in heart failure. It is unclear whether increased circulating catecholamines contribute directly to ventricular arrhythmias or are only markers of the severity of heart failure. The present study determined the sensitivity of the failing heart to the arrhythmogenic effect of exogenous norepinephrine in a rapid pacing-induced model of heart failure in dogs (240 beats for 4 wk, n = 14). A similarly operated, non paced group served as controls (n = 9). Cardiac sensitivity to the arrhythmogenic effect of catecholamines was determined by measuring the minimal dose of exogenous norepinephrine that induced ventricular tachycardia (arrhythmogenic threshold dose, ATD). ATD significantly increased after development of heart failure in heart-failure group (1.62 +/- 0.32 microgram/kg at baseline vs. 16.65 +/- 3.48 micrograms/kg at restudy, P < 0.01), whereas no significant change was noted in the control group (1.08 +/- 0.36 microgram/kg at baseline vs. 2.53 +/- 0.36 micrograms/kg at restudy, P > 0.10). Action potential duration was unchanged by superfusion with 10(-7) M isoproterenol in both ventricular muscles (230.2 +/- 6.1 vs. 229.7 +/- 5.3 ms, P = NS) and Purkinje fibers (273.2 +/- 6.5 vs. 283.8 +/ 4.2 ms, P = NS) from the failing hearts, although isoproterenol induced a shortening in the control group (204.8 +/- 0.9 vs. 181.3 +/- 1.6 ms in ventricular muscles, P < 0.01; 313.8 +/- 6.5 vs. 279.5 +/- 5.7 ms in Purkinje fibers, P < 0.01). We conclude that the failing heart has a decreased sensitivity to the arrhythmogenic effect of catecholamines. PMID- 8238577 TI - Left ventricular unloading decreases rate of isovolumic right ventricular pressure decline. AB - To determine whether direct ventricular interaction affects right ventricular (RV) relaxation, we compared the minimum rate of change of RV pressure (dP/dtmin) and the time constant, tau, of isovolumic RV pressure decline in open-chest, anesthetized pigs following a normal left ventricular (LV) contraction and an acutely unloaded LV contraction. Rapid removal of blood via the LV apex into a prosthetic ventricle during a single systole decreased the LV pressure-time integral 68.3% and peak systolic pressure 49.9% without changing RV end-diastolic conditions. Micromanometers measured ventricular chamber pressures during single RV isovolumic beats, which were produced by transient pulmonary artery occlusion in both the control and the LV-unloaded states. When the LV was unloaded, RV tau rose significantly from 41.7 +/- (SD) 7.1 to 49.8 +/- 9.4 ms, P < 0.005 (pericardium intact) and 42.9 +/- 10.8 to 51.8 +/- 13.9 ms, P < 0.005 (pericardium opened). Simultaneously, RV dP/dtmin decreased from -400.5 +/- 136.6 to -288.3 +/- 46.8 mmHg/s, P = 0.14 (pericardium intact), and -342.4 +/- 104.2 to -241.2 +/- 118.9 mmHg/s, P < 0.01 (pericardium opened). These data indicate that LV systolic unloading decreases the rate of RV relaxation. We conclude that RV relaxation is influenced by direct anatomic ventricular interaction. PMID- 8238578 TI - Leukocyte cytoskeletal structure determines capillary plugging and network resistance. AB - Leukocyte plugging in capillaries may play an important role in microvascular hemodynamics under pathological conditions that result in leukocyte activation. Previous studies have quantified the effects of transient capillary plugging on microvascular resistance for control and activated leukocytes, but have not focused on the intracellular mechanisms responsible. The frequency and duration of leukocyte-capillary plugging were measured throughout capillary networks in spinotrapezius muscle of anesthetized rats under normal physiological conditions and after leukocyte activation coupled with cytoskeletal F-actin depolymerization, using superfusion with 10(-7) M N-formyl-methionyl-leucyl phenylalanine (FMLP) and 0.3 microM cytochalasin D (CD). These data were used to estimate the increase in microvascular flow resistance due to leukocyte plugging. The increase was 1.1% in control and 0.7% in the activated FMLP-CD state and was not significantly different. CD abolished the previously reported 16% resistance increase caused by FMLP activation [Harris and Skalak, Am. J. Physiol. 264 (Heart Circ. Physiol. 31): H909-H916, 1993.], suggesting that leukocyte plugging in capillaries is primarily determined by leukocyte cytoplasmic viscosity and is not significantly affected by cell adhesion. Thus F-actin-mediated cytoskeletal reorganization during leukocyte activation has significant impact on microvascular resistance. PMID- 8238579 TI - Venoconstriction to endothelin-1 in humans: role of calcium and potassium channels. AB - Recent studies in vitro have suggested that there may be an interaction between endothelin-1 and ATP-sensitive K+ channels in vascular smooth muscle. Here we have investigated whether agents acting on membrane Ca2+ and K+ channels modulate endothelin-1-induced venoconstriction in vivo in human subjects. In a series of studies, six healthy subjects received, on separate occasions, local infusions into dorsal hand veins of endothelin-1 coinfused with 1) the ATP-sensitive K+ channel opener, cromakalim; 2) the dihydropyridine Ca2+ antagonist, nicardipine; 3) a control vasodilator, hydralazine; and 4) saline placebo. Endothelin-1 caused local venoconstriction with a maximum reduction in vein size of 66 +/- 4% at 60 min (P = 0.0001 vs. basal). Cromakalim prevented endothelin-1-induced venoconstriction (9 +/- 10% maximum constriction; P = 0.68 vs. basal). By contrast, nicardipine, in a dose sufficient to block depolarization-induced constriction caused by K+ infusion, had only a partial effect on endothelin-1 induced venoconstriction (35 +/- 8% maximum constriction; P = 0.001 vs. basal; P = 0.02 vs. endothelin-1), whereas a 10-fold higher dose of nicardipine had no additional effect and hydralazine had no effect. In further studies, cromakalim, but not nicardipine, reversed endothelin-1-induced venoconstriction. Cromakalim did not prevent constriction induced by norepinephrine. Although calcium entry through dihydropyridine-sensitive Ca2+ channels may account in part for the vasoconstrictor action of endothelin-1 in humans, the abolition of endothelin-1 responses by a K+ channel opener suggests additional mechanisms of action for endothelin-1. PMID- 8238580 TI - Cell and oxygen flow in arterioles controlling capillary perfusion. AB - Measurements of hemodynamic and oxygen transport characteristics in the arterioles that control capillary perfusion in striated muscle were used to compare oxygen flow into adjacent capillary networks. Observations were made in arterioles arising consecutively (branches 1-3 and the last branch) from a single transverse arteriole. During maximal dilation [after 5 min of exposure to superfusate with 10(-4) M adenosine (ADO)], mean cell flow into branches decreased significantly with increasing axial distance along the transverse arteriole, from 8.47 +/- 2.43 x 10(3) (SE) cells/s in branch 1 to 5.56 +/- 2.14 x 10(3), 3.21 +/- 1.30 x 10(3), and 4.00 +/- 1.33 x 10(3) cells/s in successive branches. During control, cell fluxes were not significantly different by position (2.21 +/- 1.12, 1.31 +/- 0.42, 0.97 +/- 0.31, and 1.23 +/- 0.40 cells/s in branches 1, 2, and 3 and the last branch, respectively). Branch diameters during ADO were not significantly different by position (26.2 +/- 2.9, 24.5 +/- 1.4, 22.0 +/- 2.8, and 26.7 +/- 2.7 microns, respectively). Hemoglobin saturations during ADO were not different (59.6 +/- 2.2, 60.6 +/- 2.3, 60.3 +/- 2.3, and 61.0 +/- 2.3%, respectively), whereas mean oxygen flow into branch 1 significantly exceeded that into branches 2 and 3 and the last branch (1.40 +/- 0.40 vs. 0.60 +/- 0.17, 0.53 +/- 0.22, and 0.66 +/- 0.22 pl/min, respectively). During control, oxygen flows were not significantly different between branches. Thus, both cell and oxygen flow into these arterioles vary in a systematic way dependent on their relative branch position; regulatory processes serve to make oxygen supply more uniform. PMID- 8238581 TI - Endothelin-1 exacerbates diastolic stunning in conscious dogs. AB - The purpose of this study was to examine the effects of endothelin-1 (ET-1) on "diastolic stunning" in the postischemic myocardium. In 14 conscious dogs receiving either placebo (n = 7) or ET-1 (2.5 ng.kg-1.min-1, n = 7), left ventricular (LV) hemodynamics and regional wall motion (systolic segmental shortening by sonomicrometry and the ischemic-nonischemic regional asynchrony during isovolumic relaxation) were assessed at baseline, during 10 min of left anterior descending coronary artery occlusion (CAO) and at 60 min after reflow (R 60). During CAO, the ischemic segment shortening was severely depressed and both regional asynchrony and LV relaxation time constant were significantly increased in the placebo and ET-1 groups. At R-60, this LV diastolic dysfunction recovered to baseline conditions in the placebo group but was still present in the ET-1 group. Because coronary and myocardial blood flow returned to the baseline level at R-60 in both groups, the deleterious effects of ET-1 on diastolic stunning are probably mediated by its direct action on the myocardium. PMID- 8238582 TI - Relative roles of intracellular Ca2+ and pH in shaping myocardial contractile response to acute respiratory alkalosis. AB - During acute respiratory alkalosis, myocardial contractility initially increases but then declines toward control levels. To elucidate the mechanism of this response, two parallel strategies were adopted: isovolumic left ventricular developed pressure (DP) and intracellular pH (pHi) were measured in isolated ferret hearts using 31P-nuclear magnetic resonance spectroscopy, and isometric developed tension (DT) and intracellular Ca2+ concentration ([Ca2+]i) were measured in ferret papillary muscles using microinjected fura 2 salt. When hypocapnia was induced by sudden introduction of perfusate equilibrated with 2% CO2 (from 5% CO2 in control), DP increased to a maximum of 120 +/- 3% (SE; n = 7) of control within 40 s. Afterward, DP decreased toward control levels, reaching a new steady state in 2-3 min. In contrast, pHi increased from control (7.11 +/- 0.01) only after 30 s of hypocapnia and reached a peak of 7.25 +/- 0.02 between 80 and 100 s. Thus pHi lagged behind contractility. In contrast to pHi, [Ca2+]i changed in parallel with DT: when DT reached a maximum (251 +/- 63% of control; n = 5) during hypocapnia, the amplitude of [Ca2+]i transients also peaked (190 +/- 22% of control; n = 5). A simulation of contractile force based on our measurements of pHi and [Ca2+]i, along with published Ca(2+)-tension relations, described adequately the changes in developed force during hypocapnia. These results indicate that the biphasic changes in [Ca2+]i, coupled with an out-of phase change in pHi, underlie the biphasic response of myocardial contractility to hypocapnia. PMID- 8238583 TI - An animal model of dilated cardiomyopathy: characterization of dihydropyridine receptors and contractile performance. AB - Broad-Breasted White turkey poults exhibit a low incidence of dilated cardiomyopathy (DCM). Ejection fraction in DCM turkeys with moderate to severe dilatation was decreased from 67 +/- 5% (control, n = 20) to 42 +/- 4% (DCM, n = 12; P = 0.001). Moderate to severe DCM hearts demonstrated peak left ventricular pressure 67 +/- 4 mmHg (n = 11) vs. 156 +/- 9 mmHg (n = 10; P < 0.0001). With moderate to severe dilatation, there was normal calcium responsiveness in saponin skinned fiber experiments. (+)[methyl-3H]-PN200-110 binding early in the course of cardiac dilatation was increased 62% (Bmax = 1,798 +/- 212 fmol/mg protein, n = 9 vs. control Bmax = 1,113 +/- 48 fmol/mg, n = 26; P < 0.0001). The concentration to reach 50% effect (EC50) for nifedipine contractile response was 0.5 log unit higher for isolated DCM muscle vs. control at the early stage of dilatation, consistent with the increase in calcium channel number identified by ligand binding. However, more advanced heart failure resulted in a decrease in number of DHP binding sites by 24% compared with control and by 53% compared with the early dilated group (Bmax, moderate dilatation = 844 +/- 71 fmol/mg, n = 7; P < 0.05). Finally, with gross dilatation, there was a second increase in calcium channel number by 40% (Bmax, severe dilatation = 1,556 +/- 201 fmol/mg, n = 7; P < 0.01) compared with control, which is an increase of 84% vs. moderate dilatation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238584 TI - Endothelium-dependent and -independent responses to severe hypoxia in rat pulmonary artery. AB - The changes in pulmonary artery (PA) tone that develop during hypoxia are complex and appear to involve multiple vasoregulatory pathways. This study was designed to identify the phases of the response to severe hypoxia in isolated segments of rat PA and to evaluate the mechanisms involved. Exposure of preconstricted PA segments to severe hypoxia (i.e., PO2 < 3 Torr) resulted in transient relaxation followed by vigorous hypoxic vasoconstriction (HVC), which averaged 0.26 +/- 0.02 g (or 43.9 +/- 3.7% of the maximal force developed in response to phenylephrine). HVC was followed by sustained and nearly complete inhibition of existing tone. The first two phases of the response were endothelium dependent and could be blocked by nonspecific inhibitors of endothelium-derived relaxing factor (EDRF) activity and by 500 microM N omega-nitro-L-arginine, a specific inhibitor of nitric oxide (NO) formation. The transient early relaxation could also be inhibited by superoxide dismutase (50 U/ml). In contrast, the late relaxation phase was neither endothelium dependent nor could it be blocked by inhibitors of EDRF/NO activity. Rat PA segments were relaxed by pinacidil and lemakalim, both of which activate ATP-sensitive potassium channels. Preincubation with glibenclamide, a selective inhibitor of these channels, blocked this response and also reduced late hypoxic vasodilation by nearly 40% (P < 0.001). These findings demonstrate that the response of rat PA to hypoxia involves both endothelium dependent and endothelium-independent components. The initial relaxation results from enhanced EDRF/NO activity, which is likely due to diminished breakdown by superoxide radicals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238585 TI - Elevated diaphragmatic blood flow during submaximal exercise in rats with chronic heart failure. AB - The exercise blood flow response of muscles involved in respiration was determined in rats with a myocardial infarction (MI), which was produced by tying the left main coronary artery, and in rats that underwent sham operations (Sham). Arterial blood gases, acid-base parameters, and blood flow (ml/100 g of tissue) to the diaphragm, intercostals, and transverse abdominis muscles were measured during steady-state treadmill exercise (20% grade, 28 m/min). The responses of MI rats that were classified as having a small (MIS < 25%, n = 7), medium (25% < or = MIM < or = 35%, n = 8), and large (MIL > 35%, n = 7) infarct were compared with those of Sham (n = 12) rats using analysis of variance techniques. Results demonstrated that arterial PO2 and PCO2 were similar for all groups during exercise (PaO2 = 110-112 mmHg; PaCO2 = 28-29 mmHg) even though the MIM and MIL groups had developed a significant amount of pulmonary congestion, and the MIL group demonstrated indicators of severe left ventricular dysfunction. Blood flow to the diaphragm during exercise was significantly greater for the MIL group of rats, although blood flow to the intercostals and transverse abdominis muscles was similar across the different groups. Results from this study support the contention that MI rats (including rats with decompensated heart failure) will achieve the same effective alveolar ventilation during exercise as that found for Sham rats and in the process maintain arterial O2 saturation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238586 TI - Hemodynamic responses to paraventricular nucleus disinhibition with bicuculline in conscious rats. AB - The present study was undertaken to determine the hemodynamic responses associated with stimulation of the hypothalamic paraventricular nucleus (PVN). Male Sprague-Dawley rats (n = 21) were instrumented with guide cannulas directed bilaterally at the PVN, with an electromagnetic flow probe placed on the ascending aorta and with femoral venous and arterial catheters. Bicuculline methiodide (BMI, 2 mM) was infused bilaterally (100 nl/20 min) into the PVN region before and after treatment with the beta 1-adrenergic antagonist, metoprolol bitartrate (2 mg/kg iv) or the alpha 1-adrenergic receptor antagonist, prazosin hydrochloride (2 mg/kg iv). Infusion of BMI into the PVN increased mean arterial pressure by 17 +/- 2 mmHg, and heart rate rose by 91 +/- 8 beats/min. Cardiac index increased 17 +/- 3%, whereas total peripheral resistance index was not altered significantly. After metoprolol treatment, the mean arterial pressure response to BMI was similar to control (16 +/- 2 mmHg), but the tachycardia was reduced significantly (10 +/- 4 beats/min). In addition, the blood flow response was changed qualitatively. Total peripheral resistance increased 13 +/- 3%, whereas the cardiac index response was abolished (1 +/- 2%). After prazosin treatment, BMI administration into the PVN failed to increase arterial pressure ( 1 +/- 4 mmHg). Nevertheless, the BMI infusion was associated with significant hemodynamic effects. Total peripheral resistance index decreased (-24 +/- 6%), whereas cardiac index and stroke volume index increased 34 +/- 8 and 17 +/- 5%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238587 TI - Echocardiographic volume flow and stenosis severity measures with changing flow rate in aortic stenosis. AB - The anatomy of degenerative valvular aortic stenosis has been poorly represented in animal models, limiting the evaluation of noninvasive echo-Doppler measures of transvalvular volume flow rate and stenosis severity during progressive disease evolution or under conditions of changing volume flow rates. To study these issues, chronic valvular aortic stenosis, characterized by stiff leaflets without commissural fusion, was created in nine adult mongrel dogs by suturing pericardial covered Teflon-felt pads into the sinuses of Valsalva below the coronary ostia during hypothermic cardiac arrest. In the eight surviving dogs, echo-Doppler examinations were performed weekly for up to 8 wk postoperatively. Simultaneous invasive micromanometer pressure data were collected at 2-wk intervals in all subjects, with simultaneous ascending aortic transit time-volume flow measurement in four subjects. Volume flow rates were altered with saline and dobutamine infusions during invasive studies for comparison of echo-Doppler and invasive pressure gradients, volume flow, and valve areas. Serial echo-Doppler follow-up (39 +/- 11 days) demonstrated that, from baseline to final study, mean transvalvular pressure gradient increased (4 +/- 1 to 38 +/- 7 mmHg, P = 0.001), continuity equation aortic valve area decreased (2.06 +/- 0.18 to 0.54 +/- 0.04 cm2, P < 0.0001), and progressive left ventricular hypertrophy developed (62 +/- 6 to 114 +/- 9 g, P = 0.0003). Echo-Doppler and invasive data correlated well for measures of transvalvular pressure gradients (n = 98, maximum instantaneous gradient r = 0.95, mean gradient r = 0.91), volume flow (n = 75, stroke volume r = 0.86, cardiac output r = 0.86), and valve area (n = 73, r = 0.73) despite acute alterations in volume flow and progressive disease evolution. This chronic canine model, with anatomy and hemodynamics similar to clinical degenerative valvular aortic stenosis, should provide a valuable tool for investigating clinically relevant new measures of stenosis severity with use of invasive or noninvasive techniques. PMID- 8238588 TI - Involvement of the "tethered ligand" receptor in thrombin-induced endothelium mediated relaxations. AB - The mechanisms by which the serine protease, alpha-thrombin, mediates relaxations were examined in isolated dog and pig coronary arteries and dog saphenous veins. In rings of coronary arteries and saphenous veins contracted submaximally with prostaglandin F2 alpha or U46619, alpha-thrombin (0.1-10 nM) caused relaxations that were abolished by treatment with N omega-nitro-L-arginine (L-NNA) or removal of the endothelium, indicating that the relaxations were mediated by endothelium derived nitric oxide. These relaxations were blocked by the thrombin active site inhibitor, MD-805, indicating the requirement of thrombin's catalytic site to induce the relaxations. The thrombin exosite inhibitor, BMS-180742, decreased the sensitivity to alpha-thrombin without altering maximal relaxations. Indomethacin, a cyclooxygenase inhibitor, had no inhibitory effect on the relaxations caused by alpha-thrombin, indicating that the relaxations were not mediated by cyclooxygenase products. Similar to alpha-thrombin, the thrombin receptor activating peptide (human sequence: SFLLRNP, 1-100 microM) caused relaxations in pig coronary artery and dog saphenous vein but not in dog coronary artery. These relaxations were blocked by L-NNA but not by indomethacin. The results indicate that alpha-thrombin induces endothelium-dependent relaxations by a novel signaling mechanism that involves proteolytic cleavage of the thrombin receptor to expose a new amino terminus that functions as a "tethered peptide ligand" to activate thrombin receptors on the endothelial cells and release nitric oxide. PMID- 8238589 TI - Adenosine and cerebrovascular hyperemia during insulin-induced hypoglycemia in newborn piglet. AB - The present study tested the hypothesis that adenosine is involved in mediating the hyperemic response of the newborn brain to hypoglycemia. By use of the cranial window and microdialysis-H2 clearance methodologies, changes in the diameter of pial arterioles (25-50 microns), extracellular adenosine concentrations ([ADO]), and local cerebral blood flow (CBF) were examined in isoflurane-anesthetized piglets subjected to insulin-induced hypoglycemia. Blood glucose concentrations ranged from 10 to 18 mg/dl after insulin administration (25 IU/kg iv). Local CBF in the frontal cortex increased 36 +/- 12% (P = 0.014) at 30 min of hypoglycemia (group 1, n = 12; control = 43 +/- 3 ml.min-1.100 g-1). The mean increase in dialysate [ADO] sampled concurrently from the same cortical area was 59 +/- 29% (P = 0.011; control = 0.11 +/- 0.02 microM). At 30 min of hypoglycemia, pial diameters increased 55 +/- 10% (P = 0.001; group 2, n = 9). The [ADO] in cranial window cerebrospinal fluid (CSF) increased 217 +/- 71% (P = 0.04) in response to hypoglycemia (group 3, n = 8; control = 0.016 +/- 0.006 microM). Local administration of an adenosine antagonist, 10 microM 8 sulfophenyltheophylline, to the cerebral cortex before hypoglycemia caused a 38% reduction (P = 0.011) in the pial arteriolar response at 30 min of hypoglycemia (group 4, n = 9). Similarly, local superfusion of CSF with 3.7 mM glucose attenuated the hypoglycemia-induced pial dilation 33% (P = 0.039; group 5, n = 9). Perfusion of microdialysis probes with 3.7 mM glucose in the CSF abolished the hypoglycemia-induced increase in dialysate [ADO] (group 1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238590 TI - Right and left ventricular O2 uptake during hemodilution and beta-adrenergic stimulation. AB - Myocardial O2 uptake (MVO2) and related variables were compared in right and left ventricles (RV and LV, respectively) during isovolemic hemodilution (HD) alone and combined with isoproterenol (Iso) infusion in 13 isoflurane-anesthetized open chest dogs. Measurements of myocardial blood flow (MBF) obtained with radioactive microspheres were used to calculate MVO2. Lactate extraction (Lacext) was determined. The study consisted of two experimental series: 1) graded HD (dextran) to hematocrit (Hct) of 10% and 2) Iso (0.1 microgram.kg-1.min-1 iv) during moderate HD (Hct = 18 +/- 1%). In series 1, arteriovenous O2 content difference in both ventricles decreased in parallel with reduced arterial O2 content caused by HD, i.e., percent O2 extraction was constant; MVO2 was maintained by proportional increases in MBF. In series 2, Iso during moderate HD raised MVO2 (RV, +156%; LV, +80%). Higher MVO2 was satisfied by combination of increased MBF and O2 extraction in RV and by increased MBF alone in LV. Lacext remained consistent with adequate myocardial O2 delivery throughout study. Conclusions were that 1) both RV and LV tolerated extreme HD (Hct = 10%) because blood flow reserves were sufficient to fully compensate for reduced arterial O2 content; 2) significant cardiac reserve was evident during HD, which could be recruited Iso; and 3) because increase in MVO2 in RV caused by Iso in presence of HD was partially satisfied by increased O2 extraction, the absence of augmented O2 extraction during HD alone was not due to impaired release of O2 from diluted red blood cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238591 TI - Mechanisms of volume-induced increase in left ventricular contractility. AB - Volume-induced increases in left ventricular (LV) contractility were studied in 18 anesthetized dogs. Denervation eliminated cardiac reflexes, and hearts were paced at 100 +/- 9 (SD) beats/min. Acute volume loading increased LV end diastolic pressure from approximately 4 to 14 mmHg within 1 min. Contractility increased over 10 min as measured by a decrease in end-systolic length (ESL) (sonomicrometer) at matched LV end-systolic pressure (ESP) or increase in LVESP measured at matched ESL. Volume-dependent increase in contractility was not attenuated by verapamil (0.3 +/- 0.2 microgram/kg i.v., n = 6) or myocardial stunning (15 min ischemia, 30 min reperfusion, n = 6) but was attenuated by ryanodine (1-16 micrograms/kg i.v., n = 6), which alters calcium release from the sarcoplasmic reticulum. From 1 to 10 min after volume loading, LV anterior ESL (measured at LVESP 132 +/- 12 mmHg) decreased by 6.7 +/- 0.5% before, but only by 4.0 +/- 1.7% after 1 microgram/kg ryanodine (P < 0.05). The LVESP (measured at anterior ESL 11.6 mm) increased 32 +/- 4 mmHg before, but only by 17 +/- 12 mmHg after 1 microgram/kg ryanodine. In conclusion, acute volume loading produces a time-dependent increase in LV contractility, which is mediated in part by an increase in calcium release from the sarcoplasmic reticulum. PMID- 8238592 TI - Analysis of shear stress and hemodynamic factors in a model of coronary artery stenosis and thrombosis. AB - Shear stress and alterations in blood flow within a stenosed artery promote platelet-dependent thrombosis. Using the Folts model of coronary thrombosis, we evaluated morphology, histology, and the hemodynamic properties of the stenosed vessel in 18 animals. The average stenosis created was 58 +/- 8%, with stenosed vessel diameters ranging from 0.084 to 0.159 cm. Histological examination of the stenosed vessel demonstrated that thrombi were composed primarily of platelets and formation occurred 1.0 mm downstream from the apex of the constriction, propagating distally. Peak shear stress occurred just upstream from the apex of the stenosis and varied from 520 to 3,349 dyn/cm2. Only small differences in shear forces were noted when blood viscosity was calculated using Newtonian and non-Newtonian properties. In contrast, shear stress computed for Poiseuille flow with use of the stenosis diameter underestimated the apical shear stress. Blood flow remained laminar within the stenosis with a Reynolds number range of 292 534. Our data indicate that the geometry of the stenosis inflow region must be considered in the evaluation of platelet activation and thrombus formation within a stenosed artery. PMID- 8238593 TI - Arteriolar tone is determined by activity of ATP-sensitive potassium channels. AB - The role of ATP-sensitive potassium channels (KATP) in determining resting arteriolar tone and vasodilator reactivity was assessed in superfused, hamster microcirculatory beds studied via intravital microscopy. Under resting conditions, the selective KATP blocker, glibenclamide, produced concentration dependent vasoconstriction in both the cheek pouch and the cremaster muscle. Concentration-related constriction of cheek pouch arterioles was also observed with tetrapentylammonium, although this agent appeared to have toxic effects on the microcirculation. Glibenclamide (2 microM) abolished arteriolar vasodilation to cromakalim and pinacidil over a concentration range (10 nM-1 microM) in which these agents are selective KATP agonists and also significantly inhibited adenosine-, carbacyclin-, and isoproterenol-induced vasodilation. In contrast, responses to other vasodilators were not significantly affected [methacholine, forskolin, and dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP)] or only slightly depressed (sodium nitroprusside). Thus the activity of KATP determines, in part, resting arteriolar tone in the hamster. Furthermore, vasodilators like adenosine, beta-adrenergic agonists, and prostacyclin appear to act through these ion channels by a mechanism that may not involve cAMP. PMID- 8238594 TI - Diffusion permeability of the immature placenta of the rabbit embryo to inert hydrophilic molecules. AB - The clearances of a series of hydrophilic probes (radii of 2-10 A) were determined in rabbit placentas of 13-15 days of gestation (term, 31 days). Maternal placental blood flows were measured by means of radiolabeled microspheres. None of the transfers of the tracers were limited by maternal or embryonic arteriovenous differences in the placenta, and the clearances decreased with increasing molecular radius. The transplacental water filtration rate calculated from conceptual growth was comparable in magnitude to the clearances of the largest tracers. Application of Patlak's modification of the Hertzian equation and pore theory suggested an equivalent pore radius of 17 A. Placental permeability surface area products computed from this value decreased much more steeply with increasing molecular dimension than the measured clearances, suggesting a highly significant contribution by filtration despite the very large diffusional gradients that existed under the experimental conditions. The results indicate that the size selectivity of the hemochorial embryo placenta in vivo is not very significant, and that under normal conditions filtrate of maternal plasma constitutes a major contributor to embryonic supply. PMID- 8238595 TI - Ischemic cardioprotection by ATP-sensitive K+ channels involves high-energy phosphate preservation. AB - We previously demonstrated that ATP-sensitive K+ channels (KATP) protect the guinea pig myocardium against ischemia-reperfusion injury (Cole et al., Circ. Res. 69: 571-581, 1991), but the cellular alterations leading to ischemic injury affected by KATP remain to be defined. This study investigates the relationship between activation of KATP and preservation of high-energy phosphates during global no-flow ischemia in arterially perfused guinea pig right ventricular walls. Electrical and mechanical activity were recorded via intracellular microelectrodes and a force transducer. Glibenclamide (10 and 50 microM) and pinacidil (10 microM) were used to modulate KATP. ATP and creatine phosphate (CP) levels were determined at the end of no-flow ischemia by enzymatic analysis. Preparations were subjected to 1) 20 min no-flow +/- glibenclamide (10 or 50 microM), 2) 30 min no-flow +/- pinacidil (10 microM) or pinacidil (10 microM) and glibenclamide (50 microM), or 3) 40 or 50 min of control perfusion before rapid freezing in liquid nitrogen. Pinacidil (10 microM) enhanced ischemic shortening of action potential duration (APD) and early contractile failure, prevented ischemic contracture, and inhibited high-energy phosphate depletion during ischemia. Glibenclamide (50 microM) inhibited the effects of pinacidil (10 microM) on electromechanical function and preservation of ATP and CP. Glibenclamide (10 microM) alone inhibited the early decline in APD and produced earlier ischemic contracture but did not enhance ATP or CP depletion compared with untreated tissues during 20 min of no-flow. Glibenclamide (50 microM) produced a greater inhibition of APD shortening in early ischemia, further decreased the latency to ischemic contracture, and caused enhanced ischemic depletion of ATP. The data indicate the changes in electrical activity induced by KATP indirectly preserve high-energy phosphates and reduce injury associated with ischemia. However, the data also suggest the possible presence of additional mechanisms for cardioprotection by KATP. PMID- 8238596 TI - Why does pulmonary venous pressure rise after onset of LV dysfunction: a theoretical analysis. AB - One of the most important consequences of acute left ventricular dysfunction (LVD) is pulmonary edema resulting from a rise in pulmonary venous pressure (PVP). It is generally believed that the PVP rise is a direct hemodynamic consequence of LVD. While this paradigm seems plausible, especially if the LV is viewed as a sump pump, there is no specific evidence to support this simple explanation. A theoretical analysis was performed to assess the hemodynamic mechanisms responsible for the dramatic rise in PVP after acute LVD. The ventricles were modeled as time-varying elastances; pulmonary and systemic vascular systems were modeled as series of resistive and capacitive elements. In response to a 50% decrease in LV contractile strength [end-systolic elastance (Ees)], cardiac output (CO) and mean arterial pressure (MAP) dropped substantially, while PVP increased minimally from its baseline of 12 to approximately 15 mmHg. With LV Ees set at 50% of normal, the effects of sympathetic activation were tested. When heart rate and total peripheral resistance were increased, CO and MAP improved, yet PVP still did not rise. The only intervention that caused a substantial increase in PVP was to simulate the decrease in unstressed volume (VU) of the venous system known to occur with sympathetic activation. When VU was decreased by about 15-20% (comparable to experimentally observed shifts with acute heart failure), PVP increased above 25 mmHg. The effects of pericardial constraints were investigated, and the results suggest a major role of this organ in determining the overall hemodynamic response to acute LVD, sympathetic activation, and explaining the responses to therapy. Thus this analysis suggests that elevations of PVP do not occur simply as a direct hemodynamic consequence of acute LVD. Rather, changes in PVP may be dictated more by sympathetic control on venous capacity. If confirmed, recognition of this as a primary mechanism may prove important in directing development of new therapies and in understanding the mechanisms of disease progression in heart failure. PMID- 8238597 TI - Mathematical model of blood flow in a coronary capillary. AB - The coronary capillary flow is analyzed theoretically based on continuum mechanics. The capillary is a long, elastic, and permeable vessel loaded externally by tissue pressure, and it is subject to possible periodic length changes, together with adjacent myocytes. Capillary flow is driven by arteriolar venular pressure difference. Ultrafiltration due to transmural hydrostatic and osmotic gradients is included, and consideration of mass conservation leads to a nonlinear flow equation. The results show that under physiological conditions ultrafiltration is of minor importance, and the analysis predicts regional differences in capillary flow. In regions with high tissue pressure (subendocardium), capillaries undergo significant periodic volume changes, giving rise to intramyocardial pumping. In those regions, capillary wall elasticity is of major importance. In regions with low tissue pressure (subepicardium), the possible periodic capillary length changes are predominant. The predicted flow patterns are in good qualitative agreement with measured epicardial phasic flow. In conclusion, the methodological advantage of a distributive analysis is demonstrated by its ability to elucidate and evaluate the role of flow determinants and their complex interactions. PMID- 8238598 TI - Paracrine effects of endocardial endothelial cells on myocyte contraction mediated via endothelin. AB - Endocardial endothelium is reported to modulate myocardial contraction by releasing diffusible factors, but the nature of the agent(s) responsible is unknown. In the present study we investigated the potential role of endothelin in these effects. Cultured sheep endocardial endothelial cells were found to express endothelin-1 mRNA and to release endothelin-1 into superfusing solution. This superfusate induced positive inotropic effects in isolated rat cardiac myocytes, associated with an increase in the cytosolic Ca2+ transient. Similar positive inotropic effects were induced by vascular endothelial cell superfusate as well as by synthesized endothelin-1, administered at concentrations similar to those present in the superfusate. Incubation of endocardial endothelial cell superfusate with endothelin-1-specific antiserum reduced the free endothelin-1 concentration to undetectable levels and abolished both the positive inotropic effect and the rise in cytosolic Ca2+. These findings indicate that endocardial endothelial cells may modulate myocardial contraction in part through the release of endothelin-1 and suggest that endocardial as well as vascular endothelium could exert potent paracrine effects on myocardium. PMID- 8238599 TI - Characterization of suppression of food intake following acute colon inflammation in the rat. AB - Experimental colitis was induced in rats by intrarectal infusion of trinitrobenzenesulfonic acid and ethanol. Colitis was accompanied by a large suppression of food intake of 3 days duration. The reduction of food intake was effected through a reduction of meal size, with no change in meal frequency. Those same rats demonstrating approximately 70%-80% suppression of daily food intake showed no reduction of sham feeding. These data indicate that malaise alone is inadequate to explain the suppression of food intake associated with acute colitis. Rather, the data suggest that the suppression of eating results from an exaggerated postprandial satiety signal elaborated during the period of acute inflammation, an interpretation consistent with the demonstration of a slowed rate of gastric emptying in association with the colitis. PMID- 8238600 TI - Lateral tegmental field neurons play a permissive role in governing the 10-Hz rhythm in sympathetic nerve discharge. AB - Recordings from sympathetic nerves in decerebrate cats show a variable mixture of 10-Hz and 2- to 6-Hz discharges. Although medullary lateral tegmental field (LTF) neurons are considered to be a source of the 2- to 6-Hz oscillation in sympathetic nerve discharge (SND), their role in the control of the 10-Hz rhythm has not been critically evaluated. This issue served as the focus of the current study. In the first series of experiments, spike-triggered averaging of inferior cardiac SND was used in an attempt to identify LTF neurons with activity correlated to the 10-Hz rhythm in SND. The discharges of only one of the 120 LTF neurons studied were correlated to this component of SND. In contrast, 17 of 79 neurons had activity correlated to the 2- to 6-Hz oscillation in experiments in which this component of SND was prominent. These data indicate that LTF neurons neither receive input from nor are components of the 10-Hz rhythm generator. In a second series of experiments, muscimol was microinjected into the LTF bilaterally. Chemical inactivation of the LTF either eliminated the 10-Hz rhythm or reduced the power and peak frequency in this band of SND. These data support the view that LTF neurons have a permissive role in governing the 10-Hz rhythm in SND, probably by acting on elements of the rhythm generator located elsewhere. As expected, muscimol microinjections reduced the power in the 2- to 6-Hz band in SND in some experiments. PMID- 8238601 TI - Microcalorimetric measurement of reversible metabolic suppression induced by anoxia in isolated hepatocytes. AB - The metabolic suppression due to anoxia in hepatocytes from the anoxia-tolerant turtle Chrysemys picta bellii was measured directly using microcalorimetric techniques. The normoxic heat flux from hepatocytes in suspension (25 degrees C) was 1.08 +/- 0.08 mW/g cells and decreased by 76% to 0.26 +/- 0.03 mW/g cells in response to anoxic incubation. After an acute decrease in temperature (to 10 degrees C) anoxic heat flux dropped by 96% relative to the normoxic control at 25 degrees C. The relative decrease in heat flux at both temperatures was similar, 76% at 25 degrees C and 68% at 10 degrees C. From the caloric equivalent of glycogen fermentation to lactate the heat flux from lactate production was calculated to be -93 microW/g cells (25 degrees C), and this accounted for 36% of the anoxic heat flux. When the enthalpy change associated with the release of free glucose (from glycogen breakdown) is considered, an additional 6% of the anoxic heat flux can be accounted for. Therefore, a portion of the anoxic heat flux is unaccounted for (58%), resulting in an "exothermic gap." This differs from the normoxically incubated hepatocytes where the indirect calorimetric measurement of heat flux (hepatocyte O2 consumption) could fully account for the calorimetrically measured heat flux. When normoxic hepatocytes were inhibited with cyanide, a rapid suppression in heat flux was observed. Because rapid reequilibration to a lower, cyanide-induced steady state occurred in < 15 min, it is also assumed that there is no short-term Pasteur effect in this tissue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238602 TI - Anoxic suppression of Na(+)-K(+)-ATPase and constant membrane potential in hepatocytes: support for channel arrest. AB - The maintenance of ion gradients across the plasma membrane by the Na(+)-K(+) ATPase has been shown to utilize a large fraction of the total cellular energy demand. In view of the importance of ion gradients to cellular function, and the remarkable anoxia tolerance of Chrysemys picta bellii (western painted turtle) and hepatocytes isolated from this species, it was of interest to determine if in response to anoxia 1) ion gradients were maintained and 2) if the activity of the plasma membrane Na(+)-K(+)-ATPase changed to aid in ion gradient maintenance. From normoxic hepatocyte suspensions the ouabain-inhibitable 86Rb+ uptake (a measure of Na(+)-K(+)-ATPase activity) was determined, and the rate of ATP utilization was 19.1 mumol ATP.g cells-1.h-1 or 28% of the total normoxic cellular ATP turnover. In response to anoxic incubation the activity of the pump decreased by 75% to 4.8 mumol ATP.g cells-1.h-1 and this comprised 74% of the total anoxic ATP turnover. Presently, it is not known whether the observed reduction in Na(+)-K(+)-ATPase activity is regulated by 1) allosteric modification, 2) endocytosis from the membrane, or 3) reduced Na+ influx. Plasma membrane potential was measured during anoxia, using the distribution of 36Cl-, and was not significantly different from the normoxic measurement, -30.6 +/- 3.9 and -31.3 +/- 5.8 mV, respectively. Therefore, the plasma membrane ion gradient is maintained during anoxia, and since the activity of the Na(+)-K(+)-ATPase decreases, the influx of ions must also decrease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238603 TI - Role of the brain stem in generating the 2- to 6-Hz oscillation in sympathetic nerve discharge. AB - We tested the hypothesis that brain stem circuits normally generate a 2- to 6-Hz oscillation in sympathetic nerve discharge (SND). Experiments were performed on baroreceptor-denervated decerebrate cats and urethan-anesthetized rats in which renal or splanchnic SND was recorded along with field potentials (population activity) from sites in the rostral ventrolateral medulla, medullary raphe, or medullary lateral tegmental field. Our major findings were as follows. 1) Population activity recorded from the three medullary regions contained a 2- to 6 Hz oscillation. 2) The 2- to 6-Hz oscillation in population activity recorded from some medullary sites was correlated to that in SND. Peak coherence in the 2- to 6-Hz band approached a value of 1 in some cases. 3) Whereas cervical spinal cord transection abolished or markedly reduced SND, the 2- to 6-Hz oscillation in medullary activity was essentially unchanged. These results support the view that the 2- to 6-Hz oscillation in SND can be generated in the brain stem of cats and rats. PMID- 8238604 TI - Adaptations of plasma membrane glucose transport facilitate cryoprotectant distribution in freeze-tolerant frogs. AB - Natural freeze tolerance in several anuran species involves the accumulation of high concentrations of glucose as a cryoprotectant in body fluids and tissues. The present study identifies an important new molecular mechanism supporting freeze tolerance, an adaptive increase in the capacity for facilitated transport of cryoprotectant across plasma membranes by increasing the numbers and/or activity of plasma membrane glucose transporters. Glucose transport by membranes isolated from liver and skeletal muscle was analyzed in two species, the freeze tolerant wood frog Rana sylvatica and the freeze-intolerant leopard frog Rana pipiens. Membranes from both liver and muscle of R. sylvatica displayed much higher rates of carrier-mediated glucose transport, measured by a rapid filtration technique, compared with corresponding rates for R. pipiens membranes. For the liver Vmax values for glucose transport by membrane vesicles were 69 +/- 18 and 8.4 +/- 2.3 nmol.mg protein-1.s-1 at 10 degrees C for R. sylvatica and R. pipiens, respectively. This difference was due primarily to a greater number of glucose transporters in R. sylvatica liver membranes; the total number of transporter sites, determined by cytochalasin B binding, was 4.7-fold higher in the freeze-tolerant species. For muscle membranes, the Vmax for glucose transport was 4.9 +/- 1 and 0.6 +/- 0.16 nmol.mg-1 x s-1 at 22 degrees C for R. sylvatica and R. pipiens, respectively. However, in muscle there were no differences in the number of membrane transporters between species.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238605 TI - Endotoxin stimulates drinking in rats without changing dehydrational signals controlling thirst. AB - Intravenous injections of endotoxins from Escherichia coli or Salmonella minnesota stimulate drinking and reduce urinary excretion of water and solutes in rats. E. coli endotoxin (0.15 or 0.45 mg/kg i.v.) stimulated drinking without increasing plasma osmolality or sodium concentration, hematocrit, blood hemoglobin, or plasma protein concentration and without decreasing arterial pressure. Similarly, a dipsogenic dose of S. minnesota endotoxin (0.25 mg/kg i.v.) did not reduce arterial or venous pressures or change heart rate. Blocking the renin-angiotensin system with captopril or blocking histamine receptors with pyrilamine and cimetidine did not reduce drinking or urinary fluid retention caused by E. coli endotoxin. Injections of 10 or 450 ng E. coli endotoxin into a lateral cerebral ventricle increased body temperature but not water intake. In contrast to its stimulatory effect in water-replete rats, E. coli endotoxin (0.45 mg/kg i.v.) inhibited drinking in 24-h water-deprived rats. Thus we find no evidence to support the hypothesis that endotoxin causes thirst by changing known physiological signals of cellular or extracellular dehydration. The mechanism remains unknown. PMID- 8238606 TI - Role of periaqueductal gray in the pressor response produced by central injections of angiotensin II. AB - The present studies were undertaken to determine the role of rostral periaqueductal gray (PAG) in mediating the pressor effect produced by intracerebroventricular (icv) injection of angiotensin II (ANG II, 200 ng). Two functionally and anatomically distinct sites were identified in rostral PAG: a dorsomedial site involved in the hemodynamic responses produced by electrical stimulation of the anteroventral third ventricle (AV3V) region and a ventromedial site required for the pressor response elicited by icv administration of ANG II. In Saffan-anesthetized rats, injection of lidocaine (LIDO, 4%) in dorsomedial PAG, but not in ventromedial PAG, significantly attenuated the decrease in hindquarter resistance (HQR) produced by electrical stimulation of the AV3V region, and the poststimulatory increase in mean arterial pressure (MAP) and HQR. The injection of LIDO in ventromedial PAG had no effect on the hemodynamic responses produced by electrical stimulation of the AV3V region in anesthetized rats but significantly attenuated the pressor response produced by icv administration of ANG II in conscious rats. The hypothesis that these two sites receive separate projections was addressed by microinjecting two retrogradely transported fluorescent dyes, Fluoro-Gold and Fast Blue. The anatomic findings suggest that separation of the pathways activated by electrical and chemical stimulation of the AV3V region occurs at the level of rostral PAG. PMID- 8238607 TI - Cold-defense function of brown adipose tissue during sleep. AB - Rats with chronically implanted electroencephalograph scalp electrodes and thermistors were exposed to 24 and 4 degrees C ambient temperatures during the light hours before and after acclimation to 4 degrees C ambient temperature for 9 days. During synchronized sleep, deep interscapular temperature was higher at 4 degrees C than at 24 degrees C both before and after acclimation to cold. After ablation of brown adipose tissue, deep interscapular temperature was lower at 4 degrees C than at 24 degrees C during synchronized sleep. In the presence of brown adipose tissue, deep interscapular temperature decreased sharply during desynchronized sleep at 4 degrees C both before and after acclimation to cold. This decrease was subsequent to and correlated with an increase in the temperature of the nasal mucosa. The decrease in deep interscapular temperature during desynchronized sleep at 4 degrees C ambient temperature was markedly reduced by ablation of the interscapular brown adipose tissue. PMID- 8238608 TI - Skeletal muscle as the major site of nonshivering thermogenesis in cold acclimated ducklings. AB - Despite their lack of brown adipose tissue, 6-wk-old cold-acclimated muscovy ducklings (4 degrees C; CA) exhibit nonshivering thermogenesis (NST) in the cold. To determine the site of this NST, the regional distribution of blood flow was measured by the microsphere method in the thermoneutral zone (25 degrees C) and during acute exposure to cold (8 degrees C). Ducklings reared at thermal neutrality (TN), which use shivering to produce extra heat in the cold, were compared with CA ducklings, which substitute NST for shivering. Further, the contribution of skeletal muscle thermogenesis to the increased heat production in the cold was estimated by measuring leg muscle blood flow and arteriovenous difference in oxygen content [(a-v)O2] across the leg, enabling an estimation of muscle O2 consumption. During cold exposure, a similar increase in total leg muscle blood flow occurred in TN and CA ducklings (+127 and +130% respectively), while hepatic arterial blood flow increased less (+56 to +37%, respectively). This rise in blood flow was accounted for by an increase in cardiac output, which was smaller in CA than in TN ducklings, and in both groups by a redistribution of blood flow to the most thermogenic organs (skeletal muscles and liver). The (a v)O2 across the leg was not changed by cold exposure, indicating that the increase in leg muscle O2 consumption resulted mainly from the increase in blood flow.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238609 TI - Increased role of skeletal muscle in the calorigenic response to glucagon of cold acclimated ducklings. AB - The site of the calorigenesis observed in birds after glucagon was sought in control and cold-acclimated (CA) ducklings. Twenty-four 6-wk-old muscovy ducklings were reared either at thermoneutrality (TN) (25 degrees C) or in the cold (4 degrees C) from the age of 1 wk. Glucagon-induced calorigenesis (GIC) was estimated at 25 degrees C after a peritoneal glucagon injection (103 nmol/kg). Glucagon induced a higher increase in animal heat production (indirect calorimetry) and body temperature in CA (+45% and +1.1 degrees C) than in control ducklings (+30% and +0.4 degree C). In CA ducklings, the perfusion rate (thermal clearance method) and temperature of gastrocnemius increased (+130% and +1.0 degree C) shortly after glucagon, whereas tissue oxygenation (polarography) decreased (-34%). There was no significant effect of glucagon in TN controls. These changes, which peaked 45-60 min after glucagon injection, indicated a prolonged increase of muscle O2 consumption in CA ducklings. Leg muscle blood flow (radioactive microspheres) measured 45 min after glucagon was slightly increased in controls (+20%; P < 0.05), while in CA ducklings, the increase was larger (+76%; P < 0.05). The arteriovenous difference in O2 content was not markedly affected by glucagon in both groups. These parameters indicated an increase in leg muscle O2 uptake in response to glucagon of +29% in controls and +76% in CA ducklings. In controls, 28% of the GIC measured in vivo could be attributed to whole body skeletal muscles, compared with 53% in CA ducklings. The remaining part might be accounted for mostly by the liver.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238610 TI - Plasticity of autonomic control of emesis in irradiated ferrets. AB - To investigate the plasticity of the emetic response to irradiation, five groups of ferrets (with the appropriate controls) had either the vagal or the splanchnic nerves transected and either were immediately irradiated under urethan anesthesia or were irradiated in a conscious state at 0.5-4, 4-9, and 24 h and 7 days after nerve transection. All the ferrets used in this study had the left carotid artery and the right jugular vein catheterized several hours before irradiation so that blood pressures could be monitored. In ferrets that were irradiated while anesthetized, bilateral abdominal vagotomy performed immediately before 6-Gy irradiation prevented emesis. Acute bilateral abdominal splanchnicectomy markedly reduced emesis in such cases but had no statistically significant effect in conscious ferrets. The incidence of emesis in conscious controls laparotomized 0.5-4 h before irradiation was 50% after 2 Gy and 100% after 6 Gy. Intact conscious controls were tested with 10 Gy only and showed a 100% incidence of emesis after this dose. Bilateral abdominal vagotomy, performed on ferrets 4-9 h before 8-Gy irradiation of conscious animals, markedly reduced the incidence of radiation-induced emesis. When performed 24 h before irradiation, it abolished emesis. In contrast, vagotomy was ineffective when performed 7 days before irradiation. The data suggest that, after 2-8 Gy, the vagi are the major emetic pathway in conscious ferrets and the splanchnic nerves may, under certain conditions, constitute second or alternate pathways. PMID- 8238611 TI - Alterations in body fat stores and plasma insulin levels with daily intervals of heat exposure in Holtzman rats. AB - The ability of timed daily increases in ambient temperature (from 22 +/- 1 degree C to 40 +/- 1 degree C for 2 h) to alter body fat stores, blood lipid levels, and insulin resistance were tested in male Holtzman rats. Of the six times of day tested only temperature pulses administered 16 h after light onset consistently decreased body weights, retroperitoneal fat stores, and plasma insulin levels. Subsequently, temperature pulses were administered either 0 (TP0) or 16 (TP16) h after light onset (light-dark 12:12 h). While no differences were observed between the TP0 group and the constant temperature (22 degrees C) controls, decreases in body weight gain, food consumption, retroperitoneal fat stores, and plasma concentrations of insulin, cholesterol, and triglycerides were consistently observed in the TP16 group. Although changes in plasma glucose during an oral glucose tolerance test were similar when the two treatment groups were compared with their respective controls, glucose tolerance was achieved with less insulin in the TP16 animals than in their respective controls. Insulin effectiveness was greater in the TP16 group as indicated by a decrease in plasma glucose, after insulin injection, that was of greater magnitude and longer duration than in controls. Hence, timed daily increases in ambient temperature may decrease obesity in part by decreasing plasma insulin levels apparently as a consequence of increased tissue sensitivity to insulin (greater glucose tolerance and less insulin intolerance). Because the treatment is effective only at a particular time of day the findings support a role for circadian neuroendocrine interactions in the regulation of these metabolic states. PMID- 8238612 TI - Daily intervals of heat exposure alter the phase of the daily variation of plasma hormones in Holtzman rats. AB - The effects of timed daily increases in ambient temperature (thermopulses) (from 22 +/- 1 to 40 +/- 1 degree C for 2 h) on daily variations of hormones involved in glucose and lipid metabolism were tested in male Holtzman rats (3-4 mo old) exposed to a 12-h daily photoperiod. The thermopulses were administered for 14 days either at light onset (TP0) or 16 h after light onset (TP16). Body weights and food consumption were monitored during the experiment. Retroperitoneal fat weights and plasma concentrations of insulin, glucose, glucagon, corticosterone, and prolactin were determined from blood taken every 4 h during a 24-h period commencing 24 h after the last thermopulse. TP0 treatment did not alter any of the parameters tested. Conversely, TP16 treatment obliterated the daily rhythms of insulin and corticosterone present in the controls (nonthermopulsed) and decreased body weight gains, retroperitoneal fat stores, food consumption, and the baseline levels of plasma insulin and corticosterone. The present study supports a role for circadian neuroendocrine rhythms in the reduction of fat stores induced by properly timed daily thermopulses. PMID- 8238613 TI - Role of prostaglandins in exercise-induced core temperature elevation in female Sprague-Dawley rats. AB - Female Sprague-Dawley rats (12:12-h photoperiod; body temperature, BT, measured with biotelemetry) with access to running wheels for 6 wk have an elevated BT (compared with rats with no access to exercise wheels, i.e, sedentary) both during the period of voluntary exercise (nighttime) (0.5 degree C, P = 0.0001) and the nonexercise period (daytime) (0.3 degree C, P = 0.002). To determine whether prostaglandin (PG) E was responsible for any portion of this daytime rise in BT, we injected a dose of sodium salicylate (300 mg/kg), which was shown to produce complete antipyresis in rats injected with lipopolysaccharide (LPS), into exercised and sedentary rats 4 h after the onset of the lights-on period. The injections of sodium salicylate led to a fall in body temperature in both the exercised and sedentary rats of similar amounts (-0.88 degree C vs. -0.61 degree C at 2 h postinjection, P = 0.59). We conclude that the increase in daytime BT of exercised female rats is not mediated by prostaglandins. PMID- 8238614 TI - Role of endothelium-derived nitric oxide in control of renal microvasculature in aging male rats. AB - The objective of this study was to evaluate the role of nitric oxide (NO) in the regulation of whole kidney and glomerular hemodynamics during aging. After 2 wk of oral treatment with N-nitro-L-arginine methyl ester (L-NAME; 4.5 mg.kg body wt 1.day-1) to inhibit NO synthesis, male rats, aged 3-5, 13-15, and 21-24 mo, were studied by micropuncture. Blood pressure increased by 50% in old (21-24 mo) rats with L-NAME but only 20-30% in the two younger groups. With L-NAME, renal vascular resistance increased fivefold in old rats but only twofold in younger groups. Glomerular capillary pressure increased 20-30% in younger L-NAME rats and 60% in older rats. Afferent and efferent resistances increased dramatically, and the glomerular capillary ultrafiltration coefficient decreased in all L-NAME treated rats but most strikingly in the 21- to 24-mo-old group. Acute infusion of L-arginine significantly attenuated the effects of NO synthase inhibition on arterial pressure and renal hemodynamics in both young and old rats. This study confirms that NO synthesis blockade has a greater effect on renal hemodynamics in aging rats and implies that NO may play a progressively more important role in controlling renal function with advancing age. PMID- 8238615 TI - Characterization of baroreflex impairment in conscious dogs with pacing-induced heart failure. AB - Open-loop baroreflex responses were evaluated in eight conscious dogs before and during congestive heart failure to determine the effects of failure on baroreflex control of blood pressure, heart rate, cardiac output, and total peripheral resistance. Heart failure was induced by rapid ventricular pacing. Baroreflex function was determined by calculation of the range and gain of the open-loop stimulus-response relationships for the effect of carotid sinus pressure on blood pressure, heart rate, cardiac output, and total peripheral resistance. The range and gain of blood pressure responses were substantially reduced as early as 3 days after induction of heart failure (161 +/- 6 to 99 +/- 8 mmHg and -2.7 +/- 0.3 to -1.5 +/- 0.1, respectively) and remained depressed for the 21 days of heart failure. This depression in baroreflex control of blood pressure was associated with similar depressions in reflex range and gain for heart rate (125 +/- 9 to 78 +/- 11 beats/min and -2.05 +/- 0.2 to -1.16 +/- 0.2 beats/min, respectively) and cardiac output (1.74 +/- 0.2 to 0.46 +/- 0.2 l/min and -0.81 +/ 0.02 to -0.027 +/- 0.008 l/min, respectively). The group-averaged range and gain for reflex control of vascular resistance were not altered by heart failure. In three dogs, discontinuation of rapid ventricular pacing led to resolution of heart failure within 7 days and partial restoration of the range and gain of reflex control of blood pressure. We conclude that heart failure reversibly depresses baroreflex control of blood pressure principally through a concurrent reduction in reflex control of cardiac output, whereas reflex control of vascular resistance is not consistently affected. PMID- 8238616 TI - Glucosidic pathways of glycogen breakdown and glucose production by muscle from postexercised frogs. AB - Muscle and body glucose in frogs increases markedly during the initial hour of recovery after strenuous exercise. The liver is not the major source responsible for this accumulation. This is indicated by the stability of liver glycogen levels after exercise and by the observation that hepatectomized and normal frogs accumulate similar amounts of glucose in their muscles and body during recovery. The renal contribution cannot account for this increase in body glucose. Most of the glucose that accumulates in the body after exercise has a muscular origin, as indicated by the facts that two-thirds of the body glucose is found in muscle and that the intracellular levels of muscle glucose are much higher than those of the plasma. The glucose that accumulates outside muscle may also have a muscular origin. The glucosidic pathways of glycogen breakdown are the only metabolic avenue with sufficient capacity to account for the amount of glucose accumulated in muscle during the first hour of recovery. These results indicate that the ability of an isolated preparation of frog muscle to liberate glucose during recovery from exercise (Fournier et al. J. Biol. Chem. 267: 8234-8238, 1992) is not an artifactual metabolic curiosity but rather a metabolic reality that takes place in vivo. Glucose accumulation during recovery is thought to facilitate the metabolic transition of frog carbohydrate metabolism from a catabolic state, characteristic of exercise, to an anabolic one. PMID- 8238617 TI - Sustained sleep deprivation impairs host defense. AB - Prolonged sleep deprivation in rats causes an unexplained hypercatabolic state, secondary malnutrition symptoms, and mortality. The nature of the vital impairment has long been a mystery. Its determination would help to elucidate the type of organic dysfunction that sleep prevents. There are no gross detectable disturbances in intermediary metabolism, clinical chemistry, or hematological indexes that provide substantial clues to the mediation of sleep-deprivation effects. Furthermore, postmortem examinations reveal no systematic morphological or histopathological findings. Taken together, the cachexia and the absence of evidence of structural damage or organ dysfunction pointed to involvement of a regulatory system that was diffuse, possibly the immune system. Blood cultures revealed invasion by opportunistic microbes to which there was no febrile response. These results suggest that the life-threatening condition of prolonged sleep deprivation is a breakdown of host defense against indigenous and pathogenic microorganisms. PMID- 8238618 TI - Mechanisms underlying cardiovascular defense reaction evoked by dorsal periaqueductal gray stimulation. AB - Hemodynamic and autonomic mechanisms underlying the cardiovascular defense reaction elicited by electrical stimulation of the dorsolateral periaqueductal gray region were evaluated in pentobarbital sodium-anesthetized Sprague-Dawley rats. Stimulation of this area produced transient increases in mean arterial pressure and more sustained increases in heart rate, hindlimb blood flow, and plasma catecholamine levels. The pressor component of the defense reaction was due entirely to increased total peripheral vascular resistance; elevations in cardiac output occurred only at the end of the stimulation period when blood pressure had returned to resting values. The hindlimb vasodilatory response included distinct early and late components. The late component was produced by epinephrine-mediated activation of beta 2-receptors because it was abolished by bilateral adrenalectomy or treatment with butoxamine, a beta 2-receptor selective antagonist. In contrast, the early component was only partially reduced by adrenalectomy or butoxamine treatment. During conditions of constant flow provided by a pump-perfused hindlimb preparation, the early hindlimb vasodilatory response was eliminated and a neurogenic vasoconstrictor response revealed. This finding suggests that the early hindlimb blood flow response may result from shunting of blood from highly vasoconstricted vascular beds to relatively less constricted hindlimb vasculature. PMID- 8238619 TI - Effects of temperature on muscle pHi and phosphate metabolites in newts and lungless salamanders. AB - The effect of acute alterations in body temperature (BT) on intracellular pH (pHi) and phosphate metabolites was assessed in white skeletal muscle of intact newts and lungless red-backed salamanders using 31P-nuclear magnetic resonance spectroscopy. pHi decreased with increasing BT in the tail muscle of both newts and lungless red-backed salamanders. The change in pH with change in temperature from 10 to 30 degrees C was -0.018 U/degrees C in newts and -0.041 U/degrees C in red backs. The calculated alpha-imidazole for skeletal muscle cytosol did not change (0.56) in newts from 10 to 30 degrees C but fell from 0.69 to 0.43 in red backed salamanders. Phosphocreatine (PCr)/Pi fell and Pi/beta-ATP rose with increasing temperature in both newts and red backs; however, the change was much greater in red backs. Providing the red backs with O2 at 30 degrees C led to higher pH and alpha-imidazole, comparable to that of newts, along with increased PCr/Pi and lower Pi/beta-ATP. Thus newts maintain white skeletal muscle cell cytosol alpha-imidazole constant with changes in BT, whereas red backs apparently do not. However, at the BT of preference, red backs and newts maintain similar muscle pHi and alpha-imidazole. The method of gas exchange appears to strongly influence the ability of an animal to maintain its acid-base status over a range of temperatures, and our results suggest that behavioral regulation of BT may involve alpha-imidazole regulation as well. PMID- 8238620 TI - Lateral parabrachial subnucleus lesions abolish feeding induced by mercaptoacetate but not by 2-deoxy-D-glucose. AB - Lesions of the area postrema/nucleus of the solitary tract (AP/NTS) region abolish feeding induced by mercaptoacetate (MA) and 2-deoxy-D-glucose (2DG), metabolic inhibitors that selectively impair fatty acid and glucose utilization, respectively. Because the AP/NTS region is important for both MA- and 2DG-induced feeding, the present experiment investigated the involvement of the lateral parabrachial nucleus (1PBN), which is innervated by AP/NTS neurons, in these feeding responses. Electrolytic and ibotenic acid lesions were directed at the entire parabrachial nucleus or at specific lateral parabrachial subnuclei. Rats with electrolytic lesions were tested for feeding in response to 0.9% NaCl (subcutaneous or intraperitoneal), MA (400, 600, and 800 mumol/kg ip), and 2DG (100 and 200 mg/kg sc). Ibotenate-lesioned rats were tested with NaCl and MA only. Lesions were verified either by cresyl violet staining or by glial fibrillary acidic protein immunohistochemistry. Bilateral destruction of the 1PBN severely impaired or abolished MA-induced feeding. Cell bodies important for MA induced feeding appear to be localized in the dorsal-central 1PBN subnuclear area, because both electrolytic and cytotoxin microlesions centered in this area abolished feeding in response to MA. Fibers of passage important for MA-induced feeding appear to pass through the external and superior 1PBN because electrolytic but not cytotoxin lesions of these subnuclei disrupted the feeding response. In contrast, 2DG-induced feeding did not differ significantly from sham lesioned controls in any of the 1PBN lesion groups. These results indicate that 2DG and MA stimulate feeding by activating separate central neural pathways and, perhaps, distinct metabolic controls of food intake. PMID- 8238621 TI - Fever and thermogenesis in response to bacterial endotoxin involve macrophage dependent mechanisms in rats. AB - Increases in thermogenesis and body temperature (fever) frequently accompany infection or injury and are thought to be mediated by endogenous pyrogens (e.g. cytokines), which are released from activated immune cells such as macrophages. Therefore, we have investigated the effect of selective elimination of peripheral macrophages on the changes in oxygen consumption (VO2) and colonic temperature in response to bacterial lipopolysaccharide (LPS) in the rat. Peripheral macrophages were depleted by intravenous injection of liposomes containing the drug dichloromethylene diphosphonate (Cl2MDP). Resting oxygen consumption and colonic temperatures were not affected by macrophage elimination. In intact rats, peripheral injection of LPS (0.1-0.5 mg/kg) elicited an increase in colonic temperature and in oxygen consumption that declined at higher doses (2.5 mg/kg). The pyrogenic and thermogenic responses to LPS were significantly attenuated in rats in which peripheral macrophages were eliminated. Previously, we have reported that elimination of macrophages blunts the plasma interleukin-1 (IL-1) response to LPS. Here we show that elimination of macrophages does not affect the increase in plasma IL-6 concentrations in response to LPS. These data indicate that the pyrogenic and thermogenic responses to LPS are at least in part dependent on mechanisms involving peripheral macrophages, and that peripherally produced IL-1 rather than IL-6 may be an important mediator of the changes in oxygen consumption and colonic temperature in response to LPS. PMID- 8238622 TI - Central cardiovascular effects of joining peptide in genetically hypertensive rats. AB - Joining peptide (JP) is one of the major products of proopiomelanocortin (POMC). The biological function of this peptide has not been clarified despite its relative abundance in the pituitary and the hypothalamus. Recently we demonstrated that JP, which was isolated from bovine posterior pituitary, possesses Na pump inhibitor activity. The purpose of this study is to explore the physiological relevance of JP in cardiovascular regulation. For these investigations, we used the synthetic peptides bovine JP (bJP) and COOH terminally amidated rat JP (rJP), since JP is known to have sequence variability among species. Intracisternal administration of both bJP and rJP in urethan anesthetized rats evoked similar hypertensive and tachycardia effects. The effects of both peptides were markedly greater in the spontaneously hypertensive rats (SHR) compared with the normotensive Wistar Kyoto rats (WKY). Intravenous bolus injections of rJP at the same doses were without effect. Autoradiography, using 125I-labeled [0Tyr]-rJP as a ligand, revealed specific binding sites for rJP in the dorsal medulla in areas corresponding to the nucleus tractus solitarii (NTS) (extending from approximately 0.4 mm caudal to 1.8 mm rostral to the obex). Microinjections of rJP into the caudal part of the NTS of anesthetized SHR produced dose-related pressor and tachycardic responses. The pressor and tachycardic responses were also observed at the rostral part of the NTS, whereas the injections into the intermediate part of the NTS evoked depressor and bradycardic responses in SHR. These results suggest that at doses tested, the site of JP action resides in the central nervous system, and that JP is a potent neuropeptide in medullary sites known to be pivotal in central cardiovascular regulation. The effect of JP is especially prominent in the genetically hypertensive rat. PMID- 8238623 TI - Antidipsogenic actions of endothelins are exerted via the endothelin-A receptor in the brain. AB - The receptor subtype mediating the antidipsogenic effects of the endothelins (ETs) was determined in conscious, unrestrained, normally hydrated rats. Intracerebroventricular injection of 6, 12, and 20 ng ET-2 resulted in a significant, dose-related inhibition of water drinking in response to subsequent injection of 100 pmol angiotensin II (ANG II). Pretreatment with 50 or 100 ng ET B receptor agonist failed to alter the subsequent drinking response to ANG II. Drinking in response to ANG II was significantly accentuated in rats pretreated with 50, 100, and 200 ng of the selective ET-A receptor antagonist BQ-123. These data indicate that the antidipsogenic effects of the ETs are mediated via the ET A receptor subtype and further suggest that endogenous ET plays a physiologically relevant role in the central nervous system control of fluid and electrolyte homeostasis. PMID- 8238624 TI - Do the suprachiasmatic nuclei oscillate in old rats as they do in young ones? AB - The basis of the decline in circadian rhythms with aging was addressed by comparing the patterns of three behavioral rhythms in young and old rats with the in vitro rhythm of neuronal activity in the suprachiasmatic nuclei (SCN), the primary circadian pacemaker. In some old rats, rhythms of body temperature, drinking, and activity retained significant 24-h periodicities in entraining light-dark cycles; in others, one or two of the rhythms became aperiodic. When these rats were 23-27.5 mo old they were killed, and single-unit firing rates in SCN brain slices were recorded continuously for 30 h. There was significant damping of mean peak neuronal firing rates in old rats compared with young. SCN neuronal activities were analyzed with reference to previous entrained behavioral rhythm patterns of individual rats as well. Neuronal activity from rats with prior aperiodic behavioral rhythms was erratic, as expected. Neuronal activity from rats that were still maintaining significant 24-h behavioral rhythmicity at the time they were killed was erratic in most cases but normally rhythmic in others. Thus there was no more congruence between the behavioral rhythms and the brain slice rhythms than there was among the behavioral rhythms alone. These results, the first to demonstrate aberrant SCN firing patterns and a decrease in amplitude in old rats, imply that aging could either disrupt coupling between SCN pacemaker cells or their output, or cause deterioration of the pacemaking properties of SCN cells. PMID- 8238625 TI - Influence of the renal medullary circulation on the control of sodium excretion. AB - Although the role of the renal medullary circulation in the control of urinary concentrating ability is well established, its potential influence on tubular sodium reabsorption is not generally recognized. Nearly 30 years ago, changes in the intrarenal distribution of blood flow were first proposed to contribute to the natriuretic response to volume expansion. However, the lack of reliable methods for studying medullary blood flow limited progress in this area. The recent development of laser-Doppler flowmetry and videomicroscopic techniques for the study of the vasa recta circulation has renewed interest in the role of medullary hemodynamics in the control of sodium reabsorption. Results of these studies indicate that changes in renal medullary hemodynamics alter renal interstitial pressure and the medullary solute gradient and play an important role in the natriuretic response to elevations in renal perfusion pressure, intravenous infusion of saline, and changes in tubular sodium reabsorption produced by vasoactive compounds. What is emerging from these studies is the view that changes in renal medullary hemodynamics represent an important but misunderstood and long-ignored factor in the control of tubular sodium reabsorption. PMID- 8238626 TI - Effect of superhypercapnia on cortical pHi and cortical blood flow. AB - In vivo panoramic imaging of regional cortical blood flow and intracellular brain pH with umbelliferone fluorescence was performed in fasted New Zealand White rabbits (n = 30) subjected to constant or stepwise increases in arterial PCO2 (PaCO2) up to 160 mmHg under 1.0% halothane anesthesia. At a PaCO2 of 40 mmHg, baseline brain pHi measured 7.03 +/- 0.04, while regional cortical blood flow was 47.0 +/- 4.3 ml.100 g-1.min-1. With an immediate but constant exposure to a PaCO2 of 60, 80, or 120 mmHg, at 15 min pHi fell to 6.94 +/- 0.03, 6.89 +/- 0.03, and 6.90 +/- 0.03 and recovered over 20, 80, and 120 min, respectively. At 160 mmHg PaCO2, pHi fell to 6.68 +/- 0.04 and remained acidotic. With stepwise increases in PaCO2 up to 80 mmHg, brain pHi decreased from 7.01 +/- 0.02 to 6.99 +/- 0.02, not significantly different despite a severe systemic acidosis of 7.022 +/- 0.034. At a PaCO2 of 120 and 160 mmHg, pHi dropped to 6.97 +/- 0.02 and 6.93 +/- 0.02, respectively. The difference in brain pHi between an immediate and stepwise exposure to a PaCO2 of 160 mmHg was significant (P < 0.005). These results demonstrate that brain pHi is tightly regulated across the cortical surface and resistant to profound changes in extracellular pH. The difference in pHi between the constant and incremental increases in PaCO2 suggests that there is upregulation of pH homeostatic mechanisms. PMID- 8238627 TI - Transthyretin gene expression in choroid plexus first evolved in reptiles. AB - The presence of transthyretin in mammals and birds, but not amphibia, suggested that transthyretin expression first appeared in stem reptiles. Therefore, transthyretin synthesis was studied in a lizard. Transthyretin synthesis in choroid plexus pieces from Tiliqua rugosa was demonstrated by incorporation of radiactive amino acids. Oligonucleotides corresponding to conserved regions of transthyretin were used as primers in polymerase chain reaction with lizard choroid plexus cDNA. Amplified DNA was used to screen a lizard choroid plexus cDNA library. A full-length transthyretin cDNA clone was isolated and sequenced. A three-dimensional model of lizard transthyretin was obtained by homology modeling. The central channel of transthyretin, containing the thyroxine-binding site, was found to be completely conserved between reptiles and mammals. Transthyretin expression was not detected in lizard liver. These data suggest that transthyretin first evolved in the choroid plexus of the brain. Due to a change in tissue distribution of gene expression, occurring much later during evolution, transthyretin also became a plasma protein, synthesized in the liver. PMID- 8238628 TI - Can we validate childhood psychiatric disorders in adults? PMID- 8238629 TI - Quality improvement. PMID- 8238630 TI - Oppositional defiant disorder. AB - OBJECTIVE: Oppositional defiant disorder is a common clinical diagnosis that has attracted little research interest, and doubts about its validity as a distinct category remain. However, it underwent substantial changes from DSM-III to DSM III-R, and more are proposed for DSM-IV. The objective of this study was to review the literature on this condition to establish its place in the psychiatric nosology. METHOD: The terms used in computerized searches of the literature included "oppositional disorder," "oppositional defiant disorder," and "oppositional behavior." Publications found by these searches were supplemented with references in articles, searches in the epidemiological literature, and noncomputerized searches. RESULTS: Findings of studies in which multivariate analyses were used support a distinction between oppositional defiant disorder and conduct disorder. In these studies, one-third of all community-based children with any psychiatric condition had a diagnosis of oppositional defiant disorder and used mental health services often. Symptoms of oppositional defiant disorder appear to be stable over time and to have a developmental profile and sex distribution different from those of conduct disorder. The reliability of the diagnosis is low. CONCLUSIONS: There is some support for oppositional defiant disorder as a category that reflects an oppositional-aggressive psychological dimension, which is different from a delinquent dimension. There is little evidence for making oppositional defiant disorder a part of the construct of conduct disorder and for making "lying" a criterion for it. Considerable impairment should be required for the diagnosis. A more detailed description of symptoms, including a threshold for considering them present, may increase reliability of the diagnosis. PMID- 8238631 TI - Depressive comorbidity in children and adolescents: empirical, theoretical, and methodological issues. AB - OBJECTIVE: The purpose of the study was to examine comorbidity in the context of child and adolescent depression. METHOD: The authors reviewed recent epidemiological studies using standardized interviews and DSM-III or DSM-III-R criteria. RESULTS: There was a high rate of comorbidity in children and adolescents with major depressive disorders or dysthymia. Comorbidity with conduct disorder/oppositional defiant disorder ranged from 21% to 83%; comorbidity with anxiety disorder ranged from 30% to 75%; and comorbidity with attention deficit disorder ranged from 0% to 57.1%. Rates of depressive comorbidity found in community studies were similar to the rates found in clinical studies. In almost all cases, the disorders were more common in depressed children than expected by chance, and the rates of other disorders in depressed children were higher than the rates of depression in those with depression. CONCLUSIONS: The mechanisms by which comorbidity occurs are obscure at present. Several possibilities and their implications for nosology, epidemiology, and treatment research are discussed. PMID- 8238632 TI - Patterns of psychiatric comorbidity, cognition, and psychosocial functioning in adults with attention deficit hyperactivity disorder. AB - OBJECTIVE: Although attention deficit hyperactivity disorder is a common disorder of childhood, its status as a disorder in adults is not clear. The authors reasoned that if the adult diagnosis of the disorder is a valid clinical entity, it should be similar to the childhood disorder with regard to patterns of psychiatric and cognitive findings. METHODS: Eighty-four adults with a clinical diagnosis of childhood-onset attention deficit hyperactivity disorder confirmed by structured interview who were referred for treatment were studied. Findings were compared with those from a preexisting study group of referred children with attention deficit hyperactivity disorder, nonreferred adult relatives of those children who also had attention deficit hyperactivity disorder, and adults without the disorder who were relatives of normal children. Subjects were evaluated with a comprehensive battery of psychiatric, cognitive, and psychosocial assessments. RESULTS: The referred and nonreferred adults with attention deficit hyperactivity disorder were similar to one another but more disturbed and impaired than the comparison subjects without the disorder. The pattern of psychopathology, cognition, and functioning among the adults with attention deficit hyperactivity disorder approximated the findings for children with the disorder. CONCLUSIONS: These results show that referred and nonreferred adults with attention deficit hyperactivity disorder have a pattern of demographic, psychosocial, psychiatric, and cognitive features that mirrors well documented findings among children with the disorder. These findings further support the validity of the diagnosis for adults. PMID- 8238633 TI - Quality of care for hospitalized depressed elderly patients before and after implementation of the Medicare Prospective Payment System. AB - OBJECTIVE: The authors evaluated the impact of Medicare's Prospective Payment System on aspects of quality of care and outcomes for depressed elderly inpatients in acute-care general medical hospitals. METHOD: The depressed elderly inpatients (N = 2,746) were hospitalized in 297 acute-care general medical hospitals. The authors used a retrospective before-and-after design, controlling for differences over time in sickness at admission. Quality of care and outcomes were assessed through clinical review of explicit and implicit information in the medical records; secondary data sources provided information on postdischarge outcomes. RESULTS: After implementation of the prospective payment system 1) a higher percentage of patients had clinically appropriate acute-care admissions; 2) the initial assessment of psychological status by the treating provider was more complete; 3) the quality of psychotropic medication management, as rated by the study psychiatrists, improved; 4) the rates of any inpatient medical or psychiatric complication, of discharge to another hospital or a nursing home, and of inpatient readmission declined; and 5) there was no marked change in the percentage of patients rated by study clinicians as having acceptable overall clinical status at discharge or the rate of mortality 1 year after admission. CONCLUSIONS: After the implementation of the Medicare Prospective Payment System, the quality of care for depressed elderly inpatients improved and there was no marked increase in adverse clinical outcomes. Despite these gains, after implementation the quality of care was moderate at best and over one-third of the patients had unacceptable clinical status at discharge. PMID- 8238634 TI - A comparison of psychiatric symptoms in vascular dementia and Alzheimer's disease. AB - OBJECTIVE: Psychiatric symptoms account for much of the morbidity of vascular dementia and Alzheimer's disease. The goals of this study were to extend previous observations of the psychopathology and behavioral problems associated with vascular dementia and to compare the profile of symptoms in patients with vascular dementia to that in patients with Alzheimer's disease. METHOD: Twenty eight pairs of patients (one with vascular dementia and one with Alzheimer's disease) were matched with respect to education, age, and severity of dementia. Their psychiatric symptoms were assessed with the Neurobehavioral Rating Scale, a 28-item observer-rated instrument, and the Hamilton Depression Rating Scale, and the symptoms in the two diagnostic groups were compared. RESULTS: Blunted affect, depressed mood, emotional withdrawal, motor retardation, low motivation, anxiety, unusual thoughts, and somatic concerns occurred in more than one-third of the patients with vascular dementia. There was no significant relation between severity of cognitive impairment and severity of these noncognitive symptoms. The patients with vascular dementia had more impairment than the patients with Alzheimer's disease, as indicated by the Neurobehavioral Rating Scale total scores and scores on the behavioral retardation, anxiety/depression, and verbal output disturbance factors. They also had a higher total score on the Hamilton depression scale and higher scores on 14 of the 17 Hamilton depression items. CONCLUSIONS: Patients with vascular dementia have more severe behavioral retardation, depression, and anxiety than those with Alzheimer's disease when the groups have similar levels of cognitive impairment. This probably reflects the contrasting brain regions typically involved in the two disorders. PMID- 8238635 TI - Executive impairment among the functionally dependent: comparisons between schizophrenic and elderly subjects. AB - OBJECTIVE: Executive deficits have traditionally been associated with frontal lobe brain damage. They are relevant to a variety of disabling mental conditions, including schizophrenia and Alzheimer's disease. To measure these deficits, the authors developed the Executive Interview, a 25-item, 15-minute interview. It has been validated among elderly subjects across a wide range of functional impairment. METHODS: Forty young, chronically ill schizophrenic residents of a state mental health facility and 104 elderly residents, representing three levels of care, of a comprehensive retirement community were tested with the Executive Interview and the Mini-Mental State. RESULTS: When age, gender, education, and number of prescribed medications were controlled, cognitive impairment on the Executive Interview and Mini-Mental State rose with level of care. The Executive Interview alone discriminated between subjects at each level of care, and it was more sensitive to cognitive impairment than the Mini-Mental State. Executive Interview scores correlated the strongest with level of care. Mini-Mental State scores, number of prescribed medications, and age also correlated significantly. Schizophrenic patients showed as much executive impairment on the Executive Interview as elderly subjects at the same level of care despite significant differences in age, sex, and neuroleptic use. Executive Interview and Mini-Mental State scores were highly correlated among the elderly but less so among the schizophrenic patients. Cross-group differences were also found in the pattern of failure on selected Executive Interview items despite similar total Executive Interview scores. CONCLUSIONS: Increasing executive dyscontrol is associated with the need for increasing levels of care and supervision. This finding is neither age nor disease specific. Cross-group differences on selected Executive Interview items suggest the existence of disease-specific patterns of failure. Their recognition could prove useful in the identification of anatomically or pathophysiologically distinct subgroups among patients with executive dyscontrol. PMID- 8238636 TI - Factors associated with recovery from and relapse into major depressive disorder in the elderly. AB - OBJECTIVE: This research examined demographic, clinical, and family factors in elderly depressed patients in order to ascertain which ones were related to the patients' recovery from and relapse into major depression. METHOD: One hundred twenty-seven elderly inpatients with a diagnosis of major depressive disorder according to the Research Diagnostic Criteria were evaluated at hospital admission and followed prospectively for 1 year. At the time of each patient's admission, the spouse or an adult child was also interviewed with measures that examined family-related issues. Recovery and relapse were determined according to explicit criteria from the Longitudinal Interval Follow-Up Evaluation. RESULTS: No demographic or clinical characteristics of the patients were predictors of recovery or relapse. Three family variables measured at the time of the patients' hospitalization were prospectively related to patients' status as not recovered at follow-up: the spouses' and adult children's psychiatric symptoms, their reported difficulties in caring for the depressed older patients, and their poorer physical health. However, relatives' reported difficulties with patient care were associated with reduced likelihood of relapse among the subgroup of patients who actually did recover. CONCLUSIONS: The findings are consistent with research in mixed-age samples that has documented the influence of interpersonal factors on the course of depression. PMID- 8238637 TI - Genetic and environmental contributions to dimensions of personality disorder. AB - OBJECTIVE: The authors estimated the heritability of the basic dimensions of personality disorder and the relative proportions of the variance attributable to genetic and environmental sources. METHOD: The subjects were 175 volunteer twin pairs (90 monozygotic and 85 dizygotic) from the general population. Each twin completed the Dimensional Assessment of Personality Pathology, a questionnaire that assesses 18 dimensions of personality disorder. The questionnaire was developed on the basis of factor analytic studies that identified a stable structure underlying personality disorders in clinical and nonclinical subjects. Structural equation model-fitting methods were used to estimate the influence of additive genetic, common environmental, and unique environmental effects. RESULTS: The estimates of broad heritability ranged from 0%, for conduct problems, to 64%, for narcissism. Behaviors associated with submissiveness and attachment problems had low heritability. For most dimensions, the best-fitting model was one that specified additive genetic and unique environmental effects. CONCLUSIONS: These results are similar to those reported for normal personality and suggest a continuity between normal and disordered personality. PMID- 8238638 TI - Psychopathology in the families of children and adolescents with borderline personality disorder. AB - OBJECTIVE: The purpose of this study was to determine whether rates of psychopathology differed between the families of a group of children and adolescents with borderline personality disorder and the families of a psychiatric comparison group. METHOD: The rates of psychiatric disorders in the families of 44 child and adolescent outpatients with borderline personality disorder were compared with the rates in the families of a psychiatric comparison group of 100 children and adolescents, none of whom had a diagnosis of borderline personality disorder. Psychopathology in family members was ascertained by the family history method with the use of information from semistructured interviews with the subjects in the two study groups and with their parents or guardians. The adults were systematically interviewed regarding specific DSM-III-R disorders in their families. RESULTS: The families of the patients with borderline personality disorder had significantly greater rates of psychopathology, particularly in the areas of depressive, substance abuse, and antisocial disorders. CONCLUSIONS: The finding of higher rates of psychopathology among the family members of the group with borderline personality disorder supports the hypothesis that a history of significant family psychopathology is associated with the disorder. PMID- 8238639 TI - The impact of mother-child interaction on the development of borderline personality disorder. AB - OBJECTIVE: Two major psychodynamic theories of the etiology of borderline personality disorder posit two aspects of mother-child interaction as uniquely pathogenic: maternal over-involvement with the child and mismanagement and inappropriateness of maternal guidance and support of the child. This study is an attempt to examine these putative risk factors empirically, using epidemiologic methods. METHOD: Mother-child interaction, father-child interaction, maternal personality, and adolescent diagnoses of personality disorders were measured on two occasions, 2.5 years apart, in a random sample of 776 adolescents. RESULTS: Maternal inconsistency in upbringing of the child predicted a persistence or an emergence of borderline personality disorder, but not of any other axis II disorder. However, this effect occurred only in the presence of high maternal overinvolvement. Neither maternal overinvolvement nor maternal inconsistency alone predicted emergence of borderline personality disorder. Pathological features of maternal personality did not account for the combined effect of maternal overinvolvement and inconsistency on borderline personality disorder. CONCLUSIONS: The two child-rearing risk factors hypothesized to be important by two psychodynamic models of borderline personality disorder were found to be pathogenic only when they coexisted. Their effect could not be accounted for by the biological or environmental vulnerability represented by maternal borderline personality traits. PMID- 8238640 TI - Continuation pharmacotherapy of borderline personality disorder with haloperidol and phenelzine. AB - OBJECTIVE: The aim of this study was to assess the effectiveness of low-dose neuroleptic medication and monoamine oxidase inhibitor (MAOI) antidepressant medication in continuation pharmacotherapy of patients with borderline personality disorder. METHOD: The authors conducted a double-blind, placebo controlled study comparing continuation therapy with a neuroleptic (up to 6 mg/day of haloperidol), an MAOI antidepressant (up to 90 mg/day of phenelzine), and placebo in 14 men and 40 women with borderline personality disorder. Continuation medication trials lasted 16 weeks, following 5 weeks of acute therapy. RESULTS: Continuing haloperidol demonstrated efficacy only for the treatment of irritability. Higher levels of depression, hypersomnia, and leaden paralysis were noted in the patients who received haloperidol than in those who received phenelzine and those who received placebo. The dropout rate during the first 8 weeks of the continuation study was significantly higher for the patients receiving haloperidol (64%) than for those receiving placebo (28%). Continued phenelzine demonstrated only modest efficacy for the treatment of depression and irritability. An activating effect of phenelzine was shown on measures of excitement and reactivity. CONCLUSIONS: No evidence of efficacy was found for continuation therapy with haloperidol in the treatment of borderline personality disorder other than in the treatment of irritability. Little evidence of efficacy was found for continuation therapy with phenelzine for borderline personality disorder other than modest improvements in irritability and depressive symptoms. There is currently no clear pharmacological treatment of choice for the continuation therapy of borderline personality disorder. PMID- 8238641 TI - Neuropsychological correlates of MRI temporal lobe abnormalities in schizophrenia. AB - OBJECTIVE: The authors evaluated the neuropsychological correlates of temporal lobe abnormalities in schizophrenic patients. METHOD: Fifteen schizophrenic patients underwent assessment of memory, by the Wechsler Memory Scale--Revised, and abstraction/categorization, by the similarities subtest of the Wechsler Adult Intelligence Scale--Revised and the Wisconsin Card Sorting Test. Neuropsychological tests of motor and constructional functions were used as control tasks. The patients also underwent magnetic resonance imaging (MRI) studies in which new neuroimaging techniques were used to derive measurements of volume and three-dimensional surface renderings of temporal lobe structures. RESULTS: Spearman rank-order correlations indicated significant associations between poor scores on tests of verbal memory, abstraction, and categorization and reduced volume in temporal lobe structures, including the parahippocampal gyrus and posterior superior temporal gyrus (left and right). By contrast, performance on tests of visual memory and on control tasks was not correlated with MRI temporal lobe abnormalities. CONCLUSIONS: These findings suggest a significant but modest relation between reduced volume in specific temporal lobe regions and neuropsychological deficits in abstraction, categorization, and verbal memory, all of which may reflect a dysfunctional semantic system in schizophrenia. PMID- 8238642 TI - Normalization of auditory physiology by cigarette smoking in schizophrenic patients. AB - OBJECTIVE: Because many schizophrenic patients are heavy smokers, it has been suggested that nicotine normalizes some neuronal deficit involved in their illness. Schizophrenic subjects have various difficulties with maintenance of attention and selective processing of sensory information. One defect in sensory gating in schizophrenia has been characterized by recording auditory evoked potentials. Most normal subjects have a decrease in the evoked response to the second of two closely paired stimuli, whereas most schizophrenic subjects do not. The aim of this study was to determine whether smoking normalizes this deficit in auditory sensory gating in schizophrenia. METHOD: Changes in auditory sensory gating in response to smoking cigarettes were studied in 10 smokers without psychiatric illness and 10 schizophrenic smokers. Both groups were asked to abstain from smoking from 11:00 p.m. until 8:00 a.m. the next morning, when their auditory evoked responses to pairs of clicks were recorded. The ability to gate sensory information is reflected in a decrease in the P50 wave amplitude in response to the second of the two stimuli. After baseline recordings, the subjects smoked as much as they wished, and then two postsmoking recordings were performed. RESULTS: The schizophrenic patients had a marked but brief improvement in P50 auditory gating immediately after smoking, whereas P50 gating for the normal smokers was slightly impaired. CONCLUSIONS: This study suggests that cigarette smoking can transiently normalize the impairment of auditory sensory gating in schizophrenic patients. PMID- 8238643 TI - Impaired startle prepulse inhibition and habituation in patients with schizotypal personality disorder. AB - OBJECTIVE: Information processing measures that serve as biological markers for schizophrenia can help to define the boundaries of schizophrenia. Prepulse inhibition and habituation of the blink reflex component of startle, which are thought to reflect an individual's ability to screen out or "gate" irrelevant sensory stimuli, are impaired in patients with schizophrenia. These deficits in sensorimotor gating and habituation in schizophrenic patients may lead to sensory overload and, secondarily, cognitive fragmentation, disorganization, and thought disorder. The goal of this study was to test two hypotheses: 1) patients with schizotypal personality disorder would show a loss of sensorimotor gating reflected by impaired prepulse inhibition of the human startle response and 2) patients with schizotypal personality disorder would show deficits in startle habituation consistent with the deficits seen in patients with schizophrenia. METHOD: Prepulse inhibition and habituation were assessed in 12 men and four women who met DSM-III-R criteria for schizotypal personality disorder and in 22 normal comparison subjects. RESULTS: Patients with schizotypal personality disorder had deficits in acoustic prepulse inhibition and habituation similar to the deficits observed in patients with schizophrenia. Although there were no differences between patients with schizotypal personality disorder and normal subjects in latency from acoustic startle stimuli to response, latency facilitation was produced by the prepulse in both groups. CONCLUSIONS: This pattern of changes in amplitude and latency of the startle response suggests that subjects with schizotypal personality disorder perceive the prepulse stimuli but still demonstrate deficient sensorimotor gating of amplitude. These findings support the importance of startle prepulse inhibition and startle habituation as biological markers for schizophrenia spectrum disorders. PMID- 8238644 TI - Subtypes of self-injurious patients with borderline personality disorder. AB - Twenty-seven female inpatients with borderline personality disorder were assigned to two groups on the basis of whether they did (N = 14) or did not (N = 13) report experiencing pain during self-injurious episodes. Ratings of depression, anxiety, impulsiveness, dissociation, and trauma symptoms were higher in the women who did not experience pain while injuring themselves, as were the number of suicide attempts and the prevalence of childhood sexual abuse. PMID- 8238645 TI - Trauma and posttraumatic stress disorder in subjects with anxiety disorders. AB - Trauma histories were obtained from 711 subjects in a large study of anxiety disorders, with the intent of determining the prevalence and nature of psychological trauma in this group. Twenty-seven percent of subjects reported significant trauma; 35% of these (10% of all subjects) met DSM-II-R criteria for posttraumatic stress disorder (PTSD). Subjects reporting sexual trauma were significantly more likely to have PTSD. The rate of PTSD was not higher in subjects with panic disorder than in those with other anxiety disorders. PMID- 8238646 TI - Symptoms of posttraumatic stress disorder following recovery of war dead. AB - Persons who handled human remains in Operation Desert Storm (N = 116) were compared with those who did not handle remains (N = 118) on symptoms of posttraumatic stress disorder. Subjects who handled remains reported more intrusive and avoidant symptoms than other subjects. Subjects who were inexperienced at handling remains had more symptoms than those who were experienced. Within the experienced group, there was a significant correlation between the number of remains handled and level of symptoms reported. PMID- 8238647 TI - Outcome of panic disorder with or without concomitant depression: A 2-year prospective follow-up study. AB - In a prospective 2-year follow-up study, 32 patients with panic disorder alone and 20 with panic disorder and concomitant depression were investigated. After controlled treatment with either imipramine or doxepin, patients received naturalistic treatment with antidepressants, benzodiazepines, and supportive psychotherapy. They were evaluated for anxiety, depression, and social disability at least every 3 months during the follow-up period. The data showed fluctuation of symptoms in both groups and a less favorable outcome for the patients with comorbid conditions. However, the overall outcome was better than that reported in other studies and indicates that panic disorder is quite responsive to appropriate treatment. PMID- 8238648 TI - Anxiety sensitivity and response to cholecystokinin tetrapeptide in healthy volunteers. AB - The authors determined whether fear of anxiety symptoms mediates panicogenic responses to cholecystokinin tetrapeptide (CCK-4) in healthy subjects. Individuals with a preexisting high level of anxiety sensitivity (N = 10) experienced significantly more catastrophic cognitions and fear of somatic symptoms than did subjects with low (N = 9) or medium (N = 17) anxiety sensitivity, but they were not more susceptible to experiencing a panic attack. Thus, cognitive factors do not appear to be critical determinants of CCK-4 induced panic attacks. PMID- 8238649 TI - Religious content in the DSM-III-R glossary of technical terms. AB - The authors reviewed Appendix C of DSM-III-R, Glossary of Technical Terms, for its references to religion. Religion was referred to more frequently in this glossary than it is in psychiatric research. The authors conclude that although the Glossary uses religion in constructive or cautionary reminders, the high rate of illustrative case examples of psychopathology that involve religion in the Glossary indicates cultural insensitivity in interpreting religion. PMID- 8238650 TI - Serotonin syndrome during clomipramine and lithium treatment. PMID- 8238651 TI - Olfactory hallucinations after the infusion of caffeine during sleep. PMID- 8238652 TI - Fluoxetine abuse by an intravenous drug user. PMID- 8238653 TI - Delirium following influenza vaccination. PMID- 8238654 TI - Use of nicotine patches for schizophrenic patients. PMID- 8238655 TI - Maintenance ECT for refractory obsessive-compulsive disorder. PMID- 8238656 TI - Treatment of clozapine sedation. PMID- 8238657 TI - The dangers of legalizing physician-assisted suicide. PMID- 8238658 TI - The dangers of legalizing physician-assisted suicide. PMID- 8238659 TI - The dangers of legalizing physician-assisted suicide. PMID- 8238660 TI - The dangers of legalizing physician-assisted suicide. PMID- 8238661 TI - The dangers of legalizing physician-assisted suicide. PMID- 8238662 TI - Addiction and network therapy. PMID- 8238663 TI - Borderline personality disorder and PTSD. PMID- 8238664 TI - Dysphoric or mixed mania. PMID- 8238665 TI - Dysphoric or mixed mania. PMID- 8238666 TI - Use of control groups in family studies. PMID- 8238667 TI - Failure of interpolated tests in inducing memory impairment with final modified tests: evidence unfavorable to the blocking hypothesis. AB - Whether interpolated recall or recognition tests will enhance the impairing influence of postevent misinformation on a final modified recognition test of original event information was examined in two experiments. Both used short retention intervals. Postevent information led to differences between control and misled conditions on interpolated tests, but no differences were found on modified tests. Arguments are presented that discredit the blocking hypothesis as a potential mechanism responsible for impairment found with postevent misinformation experiments that use short retention intervals. Instead, various rejection mechanisms are favorably assessed. PMID- 8238668 TI - After the flood. PMID- 8238669 TI - The bell tools for a school of public health--and for thee? PMID- 8238670 TI - Medical technology assessment--intended for whom? PMID- 8238671 TI - Adult health: a legitimate concern for developing countries. AB - Adults, defined here as people between 15 and 59 years of age, in developing countries have a high risk of premature death and suffer from frequent morbidity and high rates of chronic impairment. Their ill health imposes a major burden on health services and large negative consequences on families, communities, and societies. This paper describes the level and impact of adult mortality and morbidity, and highlights some of its characteristics and causes, which in some cases contradict commonly held beliefs. It concludes that "adult health" is a legitimate public health concern for developing countries that is not being addressed. An agenda for remedial research and action is proposed. PMID- 8238672 TI - Learning from health care experiences in developing countries. PMID- 8238673 TI - Commentary: in search of innovative approaches to international health. AB - The community health program of the Aga Khan University in Pakistan is reviewed from the perspective of historic and present-day developments in primary health care based in and focused on the community. The approach commonly referred to as community-oriented primary health care involves the complementary functions of clinical practice and epidemiology in partnership with the community. Encouragement and support by governments and other organizations for teaching and research in this field would ensure more adequate preparation of future practitioners and health scientists for community-oriented primary health care and would lead to improved health. Continuity of support for community health programs is important. PMID- 8238674 TI - A developing country's university oriented toward strengthening health systems: challenges and results. AB - OBJECTIVES: The Aga Khan University in Karachi has a mission to educate leaders and to contribute to the development of health systems for Pakistan amid challenges of scarcity and complexity. METHODS: Its key activities are (1) to design and test urban and rural health system prototypes, (2) to develop faculty in medical and nursing postgraduate community health sciences programs, and (3) to design and implement community-based undergraduate medical and nursing curricula. RESULTS: The university has developed equity-based, cost-effective primary health care prototypes in Karachi slums. With government counterparts it has tested village-, facility-, and district-level interventions in a poor rural district. Federal policymakers have taken models from each for widespread replication. The university is training 49 medical and 19 nursing faculty for postgraduate programs in community health sciences. Most faculty retain institutional leadership positions, including teaching community-based, problem solving, community health sciences as 20% of the medical and nursing undergraduate curriculum. CONCLUSIONS: The mission and experience of the Aga Khan University in population-based health systems design and health sciences education can guide universities in both developing and developed countries. PMID- 8238675 TI - Physicians who have practiced in both the United States and Canada compare the systems. AB - OBJECTIVES: The aim of this study was to examine the US and Canadian systems from the unique perspective of physicians who have practiced in both Canada and the United States. METHODS: Questionnaires were sent to 355 Canadian physicians who graduated from US medical schools and 347 US physicians who graduated from Canadian medical schools. RESULTS: The overall response rate was 59% (65% of US graduated Canadian physicians and 54% of Canadian-graduated US physicians). Thirty-six percent of the respondents were "dual experience" physicians; that is, they had practiced medicine in both countries after completing their medical training. Physicians who left Canada were more likely than those who left the United States to indicate dissatisfaction with the health care system as a reason for leaving. Respondents expressed greater professional satisfaction with their current country of practice, but overall, dual-experience physicians in the United States favored that system only slightly more than the Canadian system, whereas those in Canada rated the Canadian system significantly better than the US system. CONCLUSIONS: The comparatively weak rating of the US system by dual experience physicians underlines the need for health care reform. PMID- 8238676 TI - The effects of a natural disaster on child behavior: evidence for posttraumatic stress. AB - OBJECTIVES: A prospective study of children examined both before and after a flood disaster in Bangladesh is used to test the hypothesis that stressful events play a causal role in the development of behavioral disorders in children. METHODS: Six months before the disaster, structured measures of selected behavioral problems were made during an epidemiological study of disability among 2- to 9-year-old children. Five months after the disaster, a representative sample of 162 surviving children was reevaluated. RESULTS: Between the pre- and postflood assessments, the prevalence of aggressive behavior increased from zero to nearly 10%, and 45 of the 134 children who had bladder control before the flood (34%) developed enuresis. CONCLUSIONS: These results help define what may be considered symptoms of posttraumatic distress in childhood; they also contribute to mounting evidence of the need to develop and evaluate interventions aimed at ameliorating the behavioral and psychological consequences of children's exposure to extreme and traumatic situations. PMID- 8238677 TI - Water cost and availability: key determinants of family hygiene in a Peruvian shantytown. AB - OBJECTIVES: This study was conducted to determine whether poor hygiene practices are owing to difficulty in getting enough water and/or to ignorance of sanitary principles. METHODS: In a water-scarce shantytown in Lima, Peru, we observed in 12-hour periods over 3 consecutive days the amount of water and soap used for personal and domestic activities in 53 families and the frequency with which direct fecal contamination of hands was interrupted by washing. We also surveyed women in a similar shantytown concerning their knowledge of hygiene to ascertain whether noncompliance was owing to ignorance. RESULTS: Three hundred fecal contamination events were registered, of which only 38 (13%) were interrupted by hand washing within 15 minutes. The mean 12-hour per capita amount of water and soap used by the families was low. More than 80% of the water stored by these families had fecal coliforms. Yet the level of knowledge concerning the importance of hand washing and other hygienic practices was high. CONCLUSIONS: In water-scarce areas, sanitary education programs probably will not change hygiene practices. In these areas, an adequate supply of water is essential for good hygiene. PMID- 8238678 TI - An epidemic of pesticide poisoning in Nicaragua: implications for prevention in developing countries. AB - OBJECTIVES: The purpose of this study was to demonstrate the usefulness of the Northwestern Nicaraguan Ministry of Health surveillance system for detecting pesticide poisonings. METHODS: Cases were reported to the regional department of epidemiology through daily telephone reports and through monthly consolidated reports from each of the 18 health centers of the National Health Service. Reporting forms were also distributed to the four area hospitals. RESULTS: During June and July 1987, an epidemic of 548 pesticide poisoning was detected in northwestern Nicaragua. Seventy-seven percent of the poisonings were caused by carbofuran or methamidophos. Of the work-related cases (91% of reported poisonings), more than 80% occurred among maize farmers and on small to medium land holdings (fewer than 140 hectares). Nineteen percent of the work-related cases involved children under 16 years of age. CONCLUSIONS: Unsafe working conditions such as manual application of pesticides and the use of backpack sprayers, the introduction of a hazardous powdered formulation of carbofuran highly restricted in the developed world, and agricultural subsidies that encouraged the use of hazardous pesticides all contributed to the epidemic. PMID- 8238679 TI - Fatal occupational injury rates: Quebec, 1981 through 1988. AB - OBJECTIVES: The purpose of the study was to estimate the death rates from occupational injuries in the province of Quebec for the period 1981 through 1988. METHODS: Worker's compensation files were used to ascertain numbers of deaths, which were used as the numerators in figuring the rates (it was estimated that these files reported 83% of the true number of deaths among men). Annual average estimates of the labor force were used as denominators. RESULTS: From 1981 through 1988, compensation was awarded for 1227 fatal work injuries. Among men (96% of the victims), rates declined from 1981 to 1988 (from 12.7 to 8.1 per 100,000); women's rates were stable (< or = 1.0 per 100,000). Compared with men, women had excess mortality from violent acts. Motor vehicle crashes accounted for 36% of all fatal injuries in 1984 and 1985 and declined thereafter. Fatal injury rates in forestry and mining rose to a 1987 maximum of 67.6 per 100,000. The construction sector had the largest number of deaths, despite a decline in rates from 1981 to 1988 (from 27.8 to 15.9 per 100,000). CONCLUSIONS: Except for construction and agriculture, reported fatal occupational injury rates in Quebec were similar to those in the United States. Motor vehicle crashes, falls, violent acts, and farming-related injuries were the most frequent causes of death. PMID- 8238680 TI - Modern contraception use in Ethiopia: does involving husbands make a difference? AB - OBJECTIVES: This study was undertaken to determine the relative efficacy of home visitation with and without husband participation on the use of modern contraception in Ethiopia. METHODS: A randomized field trial of a family planning education intervention using home visitation with and without husband participation was conducted in Addis Ababa, Ethiopia, from August 1990 to December 1991 and included a 12-month postintervention follow-up. A total of 266 experimental and 261 control subjects were entered, of whom 91.7% and 88.9%, respectively, were followed through 12 months. RESULTS: A greater proportion of couples in the experimental group were practicing modern contraception at 2 months (25% vs 15%) and 12 months (33% vs 17%) following the home visit intervention. By 12 months following the home visits, experimental subjects were less likely to have defaulted and more likely to have started using modern contraception following an initial delay. CONCLUSIONS: The inclusion of husbands in family planning programs will result in relevant increases in the use of modern contraception. However, there exists an important "sleeper" effect to the education intervention, reflected by a delay of greater than 2 months in the initiation of modern contraception for most couples. PMID- 8238681 TI - Low birthweight and infant mortality in Puerto Rico. AB - OBJECTIVES: The purpose of this study was to quantify the relative contributions of maternal age, education, marital status, hospital of birth, and use of prenatal care to the high incidence of low birthweight and infant mortality in Puerto Rico. METHODS: An analysis was conducted of 257,537 live births that occurred from 1986 through 1989 among Puerto Rico residents and the 3373 corresponding infant deaths. Binomial multiple regression models were used to calculate the adjusted population attributable risks for each variable. RESULTS: Our estimates indicate that approximately 6 of every 10 infant deaths on the island are potentially preventable if low birthweight were eradicated, regardless of other associated factors. Eliminating risks associated with sociodemographic and socioeconomic factors (including hospital of birth) would potentially decrease the incidence of low birth-weight in Puerto Rico by one third. Specifically, the elimination of risks associated with the socioeconomic disadvantage of women delivering in public hospitals alone would potentially decrease Puerto Rico's low birthweight incidence by 28%, regardless of other factors considered in our study. CONCLUSIONS: Efforts to prevent low birthweight and infant mortality in Puerto Rico should focus on reducing the gap between the private and public sectors. PMID- 8238682 TI - Prescription and nonprescription drug use among black and white community residing elderly. AB - OBJECTIVES: To examine and compare concomitants of prescription and nonprescription drug use of Black and White community-dwelling elderly. METHODS: Information on prescription and nonprescription drug use, demographic and health characteristics, and use of health services was obtained from a probability-based sample of Black (n = 2152) and White (n = 1821) community-resident elderly in the Piedmont area of North Carolina. Descriptive statistics were calculated. Linear regression, in which sample weights and design effects were taken into account, was used for the final models. RESULTS: For prescription drug use, 37% and 32% of the variance was explained for Whites and Blacks, respectively (6% and 5% for nonprescription drugs). Health status and use of medical services were the strongest predictors of prescription drug use for both races (with Medigap insurance also important for Whites and Medicaid important for Blacks). Demographic characteristics and self-assessed health were significant factors in the use of nonprescription drugs. Race independently predicted use of both types of drugs but explained only a small proportion of the variance. CONCLUSIONS: Health status and use of health services are importantly related to prescription drug use. Non-prescription drug use is difficult to explain. PMID- 8238683 TI - Insurance, income, and access to ambulatory care in King County, Washington. AB - OBJECTIVES: We studied simultaneous effects of income and insurance on access measures in an indigent population, focusing on Medicaid and the marginal effects of increasing income. METHODS: Surveys were distributed in waiting rooms of county clinics and welfare offices. Models examined insurance (private, Medicaid, or none), income (to twice the poverty level), single-parent status, age, gender, and presence of a regular source of care; first-order interactions were evaluated. RESULTS: In terms of ease of access, postponing care, and having a regular source of care, uninsured respondents fared worst and Medicaid recipients were at an intermediate level. However, relative to those with private insurance, Medicaid recipients had four times the odds, and uninsured respondents twice the odds of being denied care. Income had no consistent effect; however, older, poorer people may have greater problems. For preventive services, income was significant, while differences between Medicaid and private insurance were generally not significant. CONCLUSIONS: Except for denial of care, access for indigent people is improved by Medicaid but remains worse than the access of those with private insurance. Income had variable effects, but support for income criteria used for public insurance eligibility was not found. PMID- 8238684 TI - Cancer incidence among American Indians and Alaska Natives, 1980 through 1987. AB - OBJECTIVES: This study uses Indian Health Service inpatient data to estimate cancer incidence among American Indians and Alaska Natives. METHODS: Hospital discharge data for 1980 through 1987 were used to identify cases of cancer for 21 sites in women and 18 sites in men. Estimates of incidence were directly standardized to data from the Surveillance, Epidemiology, and End Results Program for the same time frame. RESULTS: Cancers of the gallbladder, kidney, stomach, and cervix show generally high rates among many American Indian and Alaska Native communities, and cancers of the liver and nasopharynx are high in Alaska. Of the relatively common cancers in Whites, American Indians and Alaska Natives experience lower rates for cancers of the breast, uterus, ovaries, prostate, lung, colon, rectum, and urinary bladder and for leukemia and melanoma. Variation among geographic areas and among tribal groups is observed for many important cancer sites. CONCLUSIONS: This study demonstrates significant variations of cancer rates among American Indians and Alaska Natives, with important implications for Indian Health Service cancer control programs. The study also supports the potential use of hospital discharge data for estimating chronic disease among diverse American Indian and Alaska Native communities. PMID- 8238685 TI - Improving response rates through incentive and follow-up: the effect on a survey of physicians' knowledge of genetics. AB - OBJECTIVES: This study assessed efforts to increase response rates to a mailed physician survey and examined whether, as a result, nonresponse bias was reduced. METHODS: Randomly selected physicians and geneticists were mailed a questionnaire concerning genetics knowledge and attitudes. In the final but not the pilot survey, a $25 incentive and intensive follow-up were used to increase the response rate. RESULTS: The response rate from physicians in the final survey was 64.8% (n = 1140), compared with 19.6% in the pilot test (n = 69). Sample representatives in sociodemographic and practice characteristics was improved by follow-up. Respondents recruited with more difficulty did not differ on the principal outcome variable, genetics knowledge, except on one subscore. Pilot study and final survey respondents did not differ in knowledge. CONCLUSIONS: Although the effect of increased response rates on the principal outcome variable in this study was minimal, this may not be the case for other studies. Every effort should be made to attain as high a response rate as is practical and to establish that respondents are representative of the population being sampled. PMID- 8238686 TI - Weapon carrying among inner-city junior high school students: defensive behavior vs aggressive delinquency. AB - OBJECTIVES: The purpose of this study was to estimate associations between beliefs and experiences hypothesized to be related to weapon carrying among youths. METHODS: Students in two inner-city junior high schools completed anonymous questionnaires. Logistic regression models were fit for having ever carried a weapon for protection or use in a fight and were stratified by sex and weapon type. RESULTS: Among males, 47% had carried knives and 25% had carried guns. Key risk factors for knife carrying were being threatened with a knife, getting into fights, and disbelief that having a weapon increases the carrier's risk of injury. Gun carrying was associated with having been arrested, knowing more victims of violence, starting fights, and being willing to justify shooting someone. Among females, 37% had carried a knife; knowing many victims of violence and being willing to justify shooting someone predicted knife carrying. CONCLUSIONS: Knife carrying was associated with aggressiveness but did not appear to be related to serious delinquency. Gun carrying within this nonrandom sample appeared to be a component of highly aggressive delinquency rather than a purely defensive behavior. PMID- 8238687 TI - Young men's risk behaviors for HIV infection and sexually transmitted diseases, 1988 through 1991. AB - OBJECTIVES: This paper analyzes age- and period-related changes in risk behaviors for human immunodeficiency virus (HIV) infection and sexually transmitted diseases among young men in the United States between 1988 and 1991. METHODS: Data were from the 1988 and 1991 waves of the National Survey of Adolescent Males. The 1988 survey was a nationally representative survey of 1880 males aged 15 through 19 years. The 1991 survey was a longitudinal follow-up of 1676 males aged 17 through 22 years. RESULTS: As they aged, the young men increased their levels of sexual activity and decreased their condom use. Period-related changes between 1988 and 1991 were examined by comparing similar cohorts of 17.5- through 19-year-old men: there were signs that sexual activity and intravenous drug injection increased, but condom use did not change significantly. In 1991 51% of the young men said they were occasionally "high" during sex, a state that is related to reduced condom use. CONCLUSIONS: Early progress in fostering safer behaviors among young men slowed and possibly stopped as the nation entered the 1990s. Prevention efforts need to be renewed and should focus on older youth and young adults. PMID- 8238688 TI - Anthropometry and childhood mortality in northwest and southwest Uganda. AB - Two longitudinal studies were carried out in northwest and southwest Uganda to examine the relationship between anthropometry and childhood mortality. Although the prevalence of malnutrition was significantly different between the two geographic areas, the relative risk for mortality associated with low levels of anthropometry was similar. When the anthropometric parameters were compared among each other, mid-upper arm circumference was found to be the most powerful predictor of mortality. The findings of this study confirm that mid-upper arm circumference is the indicator of choice to identify children at higher risk of death. PMID- 8238689 TI - The authorship and fate of international health papers submitted to the American Journal of Public Health in 1989. AB - We reviewed the authorship characteristics, editorial processing, and final fate of 126 papers dealing with data from countries other than the United States and Canada and submitted to the American Journal of Public Health in 1989. The acceptance rate of these international health papers was 22%, similar to that of all papers (25%). Authors from developed countries had higher acceptance rates than authors from developing countries, but the highest acceptance rate (36%) was for international health papers with joint authorship from both developed and developing countries. Of 83 rejected papers, 72% were published in other journals. Of these, 45% were published in journals covered by Index Medicus, a figure similar to that for all papers rejected by the Journal. PMID- 8238690 TI - Screening for colorectal neoplasia: physicians' adherence to complete diagnostic evaluation. AB - This prospective study was done in a health maintenance organization colorectal cancer screening program to determine whether 166 persons found to have abnormal fecal occult blood test results typically underwent complete diagnostic evaluation (i.e., either colonoscopy or barium enema x-ray plus flexible sigmoidoscopy). Chart audit data show that 137 (82%) subjects contacted a physician to discuss follow-up. A complete diagnostic evaluation was recommended to only 52 (38%) patients who talked with a physician. Forty-two (81%) patients who were advised to get a complete diagnostic evaluation actually complied. Significant differences in clinical findings were observed for patients who did and did not have a complete diagnostic evaluation. PMID- 8238691 TI - The epidemiology of child abuse: findings from the Second National Incidence and Prevalence Study of Child Abuse and Neglect. AB - The epidemiology of child abuse was investigated with data from the Second National Incidence and Prevalence Study of Child Abuse and Neglect. A statistical comparison of incidence rates suggested that age, family income, and ethnicity were risk factors for both sexual abuse and physical abuse, but county metrostatus was not. Gender was a risk factor for sexual abuse but not for physical abuse. A logistic regression analysis showed that ethnicity, county metrostatus, and a gender-by-income interaction distinguished sexual abuse from physical abuse. PMID- 8238692 TI - Activity, inactivity, and obesity: racial, ethnic, and age differences among schoolgirls. AB - Physical activity, inactivity, and obesity were assessed by questionnaire among a multiethnic sample of 552 girls in grades 5 through 12. Hispanics and Asians reported lower activity levels than other racial groups. Only 36% of the entire sample and less than one fifth of either Asians or Hispanics met the year 2000 goal for strenuous physical activity. Physical activity was inversely associated with age and age-adjusted body mass index. Obesity was unrelated to inactivity. PMID- 8238693 TI - Safety behavior among Iowa junior high and high school students. AB - The goal of this study was to determine demographic factors associated with reported safety behavior by studying 2250 Iowa junior high and high school students via a self-administered questionnaire. Students attending rural schools used front seat belts and helmets less frequently than urban students. Seat belt and helmet use and swim safety decreased dramatically with age. Occurrences of driving or riding while drunk or high increased with age. Boys were less likely than girls to wear back seat belts and moped helmets and to check water depth before diving. Possession of a driver's license was not independently associated with any of the safety behaviors. PMID- 8238694 TI - The choice of weapons in firearm suicides in Iowa. AB - This study reports the types of firearms used in firearm suicides in Iowa and tests the hypothesis that the use of handguns, rifles, and shotguns in such suicides reflects the availability of these weapons. The percentage of firearm suicides involving handguns increased from 36.6% in 1980 through 1984 to 43.3% during 1990 and 1991. In both time periods, the use of handguns was more likely than would be expected from their household prevalence. Our data suggest that handguns are disproportionately represented among firearm suicides and that this overrepresentation has increased during the last decade. PMID- 8238695 TI - The effects of resistance strategies on rape. AB - This investigation sought to clarify which resistance strategies were associated with rape avoidance and at what cost. Data were gathered from initial and supplemental police reports about 150 sexual assault victims, ages 16 and older, who were assaulted between June 1, 1988, and May 31, 1989, in Omaha, Neb. Although this analysis could not determine causality, it did indicate that forceful verbal resistance, physical resistance, and fleeing were all associated with rape avoidance, whereas nonforceful verbal resistance and no resistance were associated with being raped. Further, women who used forceful resistance were no more likely to be injured than women who did not resist. PMID- 8238696 TI - Medical technology assessment and practice guidelines: their day in court. AB - There is the expectation that outcomes research and the promulgation of medical practice guidelines will be able to identify and hopefully reduce the amount of unnecessary or inappropriate medical care through a variety of methods, including utilization review. However, past efforts by public and private insurers to deny claims on the basis of formal technology assessments or practice guidelines have frequently been overturned by the courts for multifarious reasons. This paper examines the court's reluctance to accept a variety of technology assessment methods in coverage policy decisions. The paper reviews the options that have been proposed to restrict judicial involvement in the formulation of coverage policy and then proposes a new option that employs a more precise taxonomy of medical practice assessment. PMID- 8238697 TI - The prevalence of smoking among physicians in Japan. PMID- 8238698 TI - Risk of HIV infection among Army Reserve Components physicians. PMID- 8238699 TI - Misuse of pediatric emergency services and fathers' employment status. PMID- 8238700 TI - Firearm deaths in the United States and gun availability. PMID- 8238701 TI - Protecting construction workers from exposure to lead: success in New Jersey. PMID- 8238702 TI - Spear tackler's spine. An entity precluding participation in tackle football and collision activities that expose the cervical spine to axial energy inputs. AB - We describe spear tackler's spine, a clinical entity that constitutes an absolute contraindication to participation in tackle football and other collision activities that expose the cervical spine to axial energy inputs. A subset of football players were identified who demonstrated: 1) developmental narrowing (stenosis) of the cervical canal; 2) persistent straightening or reversal of the normal cervical lordotic curve on erect lateral roentgenograms obtained in the neutral position; 3) concomitant preexisting posttraumatic roentgenographic abnormalities of the cervical spine; and 4) documentation of having employed spear tackling techniques. From data obtained by the National Football Head and Neck Injury Registry and the senior author's practice, 15 cases of spear tackler's spine were identified during 1987 to 1990. All 15 cases were evaluated because of complaints referable to the cervical spine or brachial plexus resulting from football injuries. Of these, 11 had complete neurologic recovery without permanent sequelae. Four cases resulted in permanent neurologic deficits: quadriplegia, 2; incomplete hemiplegia, 1; and residual long track signs, 1. Permanent neurologic injury occurred as the result of axial loading of a persistently straightened cervical spine from use of head-impact playing techniques. We suggest that individuals who possess the aforementioned characteristics of spear tackler's spine be precluded from participation in collision activities that expose the cervical spine to axial energy inputs. PMID- 8238703 TI - Upper trunk brachial plexopathy. The stinger syndrome. AB - This study was designed to determine the cause of upper trunk brachial plexopathy, which is referred to as a "stinger" or a "burner." This injury often has been thought to occur secondary to traction when an athlete sustains a lateral flexion injury of the neck. At the United States Military Academy, a 4 phase study was begun with 261 tackle football players (236 intramural- and 25 varsity-level players) to investigate this injury. Electromyography and nerve root stimulation studies were used to delineate the lesion, which was found in a total of 32 players who continued throughout the study. This study demonstrated that a much more common mechanism of injury resulting in the stinger syndrome is probably compression of the fixed brachial plexus between the shoulder pad and the superior medial scapula when the pad is pushed into the area of Erb's point, where the brachial plexus is most superficial. An orthosis was designed to protect the brachial plexus from the compressive force of the shoulder pad. In preliminary trials, this orthosis had been very effective in decreasing the number of episodes in which stinger injuries occurred. PMID- 8238704 TI - Arthroscopic transglenoid suture capsulolabral repairs: preliminary results. AB - The arthroscopic transglenoid suture technique was performed on 38 shoulders for anterior capsulolabral repair or reconstruction. The primary complaint was instability in 34 shoulders (89%) and pain in 4 shoulders (11%). In the instability subgroup, 3 (9%) experienced instability in their sleep, 17 (50%) with activities of daily living, and 14 (41%) with athletic activities. Arthroscopic examination revealed labral detachment in 35 shoulders (92%) with additional capsular abnormalities noted in 17 (45%). The remaining 3 shoulders (8%) demonstrated capsular laxity and thinning without labral detachment. Twenty nine shoulders had complete relief of instability. There were no redislocations. Four shoulders (10%) had recurrence of instability. Twenty shoulders (53%) obtained full range of motion, 15 (39%) had minor (< 10 degrees) loss of external rotation, 2 (5%) experienced greater (> 10 degrees) loss of external rotation, and 1 improved over a restricted preoperative range of motion. Fifteen of the 20 competitive athletes and 11 of the 15 recreational athletes returned to the same level and same type of athletic activity. Five patients did not resume their preinjury athletics because of unrelated life-style changes; they reported no shoulder problems. Four patients significantly reduced their athletic participation because of postoperative instability or residual pain. In conclusion, relief of apprehension, reestablishment of shoulder stability and return to athletic activity, including contact and throwing activities, can be achieved safely and effectively in appropriately selected patients. PMID- 8238705 TI - Long-term followup of the modified Bristow procedure. AB - A total of 86 modified Bristow procedures were performed for anterior shoulder instability between 1975 and 1987. Followup on 79 shoulders (92%) was obtained at an average postoperative time of 8.6 years. The redislocation rate was 4%. Average motion loss was 5 degrees of internal rotation and 9 degrees of external rotation. Fifteen percent of the patients examined expressed mild apprehension with the shoulder abducted and externally rotated. Radiographic bone union of the coracoid transplant was noted in 82% of patients. Additional surgical procedures were required in 14% of patients. Seventy-three percent of the reoperations were for screw removal because of persistent shoulder pain. The average subjective shoulder function was rated at 86% of preinjury level. All throwing athletes were able to return to throwing, although 54% of the patients with dominant shoulder involvement noted a decrease in throwing velocity. Ninety-seven percent of the patients rated their results as good or excellent. PMID- 8238706 TI - The natural history of meniscal tears in anterior cruciate ligament insufficiency. AB - We reviewed the meniscal status of 176 consecutive patients undergoing anterior cruciate ligament reconstruction acutely (less than 6 weeks from injury), subchronically (6 weeks to 12 months from injury), and chronically (more than 12 months from injury). The commonest tear was the single longitudinal vertical split of the medial meniscus. There was an increasing incidence of meniscal tears as the injury became more chronic, with a significant (P < 0.001) increase in medial meniscal tears; the incidence of lateral meniscal tears remained relatively constant. Seventy-five (43%) of the patients had one or both menisci repaired. Acutely, repair was performed more frequently on the medial meniscus than the lateral (80% versus 24%, respectively). All repaired menisci had single longitudinal tears unstable to probing. The incidence of repair dropped to 46% in the medial meniscus and 14% in the lateral meniscus in the chronic stage. Nineteen (25%) of these 75 patients (26 menisci) underwent a check arthroscopy at a minimum of 6 months from repair. All 21 medial menisci and all 5 lateral meniscal tears had healed; however, 1 lateral meniscus had torn along the line of the sutures. At an average followup of 40 months, 92% of the repaired menisci were still in situ and 8% that had required resection were related to the recurrence of anterior cruciate ligament instability. This study highlights the increasing incidence of meniscal injury in chronic anterior cruciate ligament insufficiency with the meniscal tears becoming more complex and therefore less amenable to suture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238707 TI - Comparison of arthroscopic and open reconstruction of the anterior cruciate ligament. Early results. AB - The purpose of this study was to determine in a prospective, randomized, blinded design whether arthroscopically assisted anterior cruciate ligament reconstruction offered any significant immediate or short-term advantages over traditional open reconstruction through a limited arthrotomy. Patients with a diagnosis of deficiency of the anterior cruciate ligament were randomly assigned to one of two treatment groups: the open group (limited open reconstruction) or the arthroscopic group (fully arthroscopic reconstruction). Postoperatively, both groups were treated identically. Intra- and postoperative observations included length of surgery, duration of hospitalization, and amount of pain medication. Follow-up evaluations were performed at 1, 6, 12, 16, 20, and 24 weeks to record crepitus, swelling, range of motion, ligament laxity, and thigh atrophy. Lysholm scores were obtained at the 16 and 24 week follow-ups. At 24 weeks, 86% of the open group and 89% of the arthroscopic group had good-to-excellent results. Intraoperative, postoperative, and follow-up findings indicated no statistically significant differences or relationships between the two groups in any of the variables measured, except that operative time was 13 minutes longer in the arthroscopic group (P < 0.001). The results do not substantiate a clinical advantage for either technique. PMID- 8238708 TI - Intraarticular anterior cruciate ligament reconstruction in the symptomatic arthritic knee. AB - We reviewed a consecutive series of young symptomatic patients with chronic anterior cruciate ligament-deficient knees to determine if an autogenous patellar tendon graft reconstruction decreased their symptoms and increased the stability of the knee. All patients had radiographic evidence of posttraumatic arthritis. Thirty-three patients met our criteria for inclusion in the study. Time from injury to reconstruction of the anterior cruciate ligament averaged 105 months. All patients underwent an accelerated rehabilitation program designed to help them regain full range of motion as soon as possible. Preoperative and postoperative range of motion, strength, stability, and subjective evaluations were compared. Followup averaged 44.8 months. Follow-up range of motion was not significantly different from preoperative measurements (P = 0.51). Postoperative stability, as measured by KT-1000 arthrometer maximum manual test, averaged 2.7 mm versus 8.4 mm preoperatively (P < 0.001). Isokinetic testing revealed no difference in the quadriceps strength after reconstruction (P = 0.99). Patients' subjective scores on a modified Noyes questionnaire improved for pain, stability, and activity level, with a total improvement from 55 to 81 points. Although the procedure and rehabilitation were successful in providing stability and decreasing pain, patients were still encouraged to limit high-impact athletic and occupational activity. PMID- 8238709 TI - Does Cybex testing increase knee laxity after anterior cruciate ligament reconstructions? AB - Long-term stability of the knee after anterior cruciate ligament reconstruction is imperative. Testing protocols that use isokinetic systems are commonly performed despite controversies as to their safety. The purpose of this study was to test whether one episode of isokinetic testing would cause an increase in anterior tibial translation. Twenty-four subjects who had anterior cruciate ligament reconstructions 153 to 300 days earlier volunteered for the study. Initially, subjects walked on a treadmill at 5 km/hr for 10 minutes to test the effect of exercise on displacement measurements. One week later, subjects performed a maximal knee flexion-extension test on the Cybex dynamometer at 60, 150, and 240 deg/sec. Anterior tibial displacement at 133.5 N was obtained from force-displacement curves produced by KT-2000 arthrometer testing at nine intervals: before exercise on the treadmill, at four intervals after treadmill exercise, and at four intervals after Cybex testing. Repeated measures analysis of variance did not show a significant exercise effect, interaction between type of exercise and time interval, or change after Cybex testing for the reconstructed knee displacement, the contralateral knee displacement, or side-to side difference. The average difference before and after Cybex testing was 0.1 mm for the reconstructed knee. In conclusion, a single Cybex test, performed at least 6 months after surgery, did not affect anterior tibial displacement in this study sample. PMID- 8238710 TI - Injuries in professional rugby league. A three-year prospective study of the South Sydney Professional Rugby League Football Club. AB - In this 3-year prospective study, the incidence and nature of injuries incurred by a professional rugby league football club were investigated. During the 1989, 1990, and 1991 season games, 141 injuries occurred throughout the first, second, and under-21 age teams, which resulted in players missing subsequent games. The incidence of injury was 44.9 per 1000 player-position game hours, which is high when compared with other sports. Of these injuries, 37.6% were classified as minor, 34.8% as moderate, and 27.6% as major. The classification was based on the number of subsequent games missed: minor injuries caused a player to miss one game; moderate, two to four games; and major, five or more games. Ligament and joint injuries comprised 53.9% of all injuries, and the knee was the most common area injured (24.1%). The commonest specific injuries were to the medial collateral ligament of the knee and to the groin musculotendinous unit (10.6% each). PMID- 8238711 TI - Australian snowboard injury data base study. A four-year prospective study. AB - Information on the rate and spectrum of snowboarding injuries is limited. This 4 year prospective study at 3 major Australian ski resorts assesses incidence and patterns of snowboarding injuries, particularly in relation to skill level and footwear. Ski injury data were collected for the same period. In a predominantly male study population (men:women, 3:1), 276 snowboarding injuries were reported; 58% occurred in novices. Fifty-seven percent of injuries were in the lower limbs, 30% in the upper limbs. The most common injuries were sprains (53%), fractures (24%), and contusions (12%). Comparing skiers' versus snowboarders' injuries, snowboarders had 2.4 times as many fractures, particularly to the upper limbs (21% versus 35% of upper limb injuries), fewer knee injuries (23% versus 44% of lower limb injuries), but more ankle injuries (23% versus 6% of lower limb injuries). Ankle injuries were more common with soft-shell boots, worn most by intermediate and advanced riders. Knee injuries and distal tibial fractures were more common with hard-shell boots, worn most by novices. Overall, novices had more upper limb fractures and knee injuries; intermediate and advanced riders had more ankle injuries. Falls were the principal mode of injury. To prevent injury, beginners should use "hybrid" or soft-shell boots and take lessons. PMID- 8238712 TI - Intrinsic risk factors for exercise-related injuries among male and female army trainees. AB - Physical training-related injuries are common among army recruits and other vigorously active populations, but little is known about their causation. To identify intrinsic risk factors, we prospectively measured 391 army trainees. For 8 weeks of basic training, 124 men and 186 women (79.3%) were studied. They answered questionnaires on past activities and sports participation, and were measured for height, weight, and body fat percentage; 71% of the subjects took an initial army physical training test. Women had a significantly higher incidence of time-loss injuries than men, 44.6% compared with 29.0%. During training, more time-loss injuries occurred among the 50% of the men who were slower on the mile run, 29.0% versus 0.0%. Slower women were likewise at greater risk than faster ones, 38.2% versus 18.5%. Men with histories of inactivity and with higher body mass index were at greater injury risk than other men, as were the shortest women. We conclude that female gender and low aerobic fitness measured by run times are risk factors for training injuries in army trainees, and that other factors such as prior activity levels and stature may affect men and women differently. PMID- 8238713 TI - Prevention of running injuries by warm-up, cool-down, and stretching exercises. AB - The purpose of this study was to evaluate the effect of a health education intervention on running injuries. The intervention consisted of information on, and the subsequent performance of, standardized warm-up, cool-down, and stretching exercises. Four hundred twenty-one male recreational runners were matched for age, weekly running distance, and general knowledge of preventing sports injuries. They were randomly split into an intervention and a control group: 167 control and 159 intervention subjects participated throughout the study. During the 16-week study, both groups kept a daily diary on their running distance and time, and reported all injuries. In addition, the intervention group was asked to note compliance with the standardized program. At the end of the study period, knowledge and attitude were again measured. There were 23 injuries in the control group and 26 in the intervention group. Injury incidence for control and intervention subjects was 4.9 and 5.5 running injuries per 1000 hours, respectively. The intervention was not effective in reducing the number of running injuries; it proved significantly effective (P < 0.05) in improving specific knowledge of warm-up and cool-down techniques in the intervention group. This positive change can perhaps be regarded as a first step on the way to a change of behavior, which may eventually lead to a reduction of running injuries. PMID- 8238714 TI - Outpatient percutaneous screw fixation of the acute Jones fracture. AB - Nine patients (8 men and 1 woman, ranging in age from 17 to 22 years) who sustained a Jones fracture were treated with percutaneous intramedullary screw fixation as outpatients. All of the patients were varsity athletes. Seven were Division I scholarship athletes. Beginning at 7 to 10 days after surgery, all patients were allowed weightbearing as tolerated with a CAM walker. Stationary bicycling, swimming, and Stairmaster were allowed at 2 to 3 weeks. The average return to running was 5.5 weeks (range, 3 to 10). The average return to full competition was 8.5 weeks (range, 7 to 12). No perioperative or postoperative complications occurred. Average followup was 2.5 years. All fractures attained clinical and radiographic union. We believe that outpatient percutaneous intramedullary screw fixation of the acute Jones fracture is a reasonable alternative for those active patients who would have difficulty with a non weightbearing cast and crutches or who desire an expeditious return to activities. Time restraints are particularly critical for in-season or preseason athletes. With the outpatient screw fixation method, our patient population had predictable healing, and they returned to full sports participation within 12 weeks. PMID- 8238715 TI - Percutaneous Achilles tendon repair. AB - Fourteen patients underwent percutaneous Achilles tendon repairs between 1982 and 1989 for ruptures approximately 2 to 8 cm from the calcaneal insertion. They were evaluated subjectively (questionnaires) and objectively (physical examinations, Cybex II dynamometer). The minimum follow-up time was 2 years with an average of 3.8 years. Subjectively, all of the patients were satisfied with their overall results. Objectively, no significant difference was demonstrated by Cybex II evaluation at 60 and 120 deg/sec. A significant difference was seen at 180 deg/sec, but this represented only a 13% loss of power. No infections, delayed wound healing, reruptures, or evidence of thrombophlebitis were noted. There was one sural nerve injury. PMID- 8238716 TI - The quantitative measurement of normal passive medial and lateral patellar motion limits. AB - To quantify normal motion, medial and lateral passive patellar motion limits were measured in 67 high school athletes randomly selected from a group of 1340 athletes undergoing preseason physical examinations. Patellar displacement was measured at knee flexion angles of 0 degree and 35 degrees, using both a Patella Pusher (a hand-held force gauge) and a manual technique, and the results were compared. Demographic data and physical examination of the deceleration mechanism (Q angle, vastus medialis obliquus dysplasia, patella alta and baja, and valgus and varus alignment) were correlated with patellar motion limits. With the knee in extension, passive displacement of the patella averaged 9.6 mm medially and 5.4 mm laterally. In flexion, medial displacement averaged 9.4 mm and lateral displacement averaged 10.0 mm. No positive correlations were found between demographic data or deceleration mechanism examination parameters and patellar motion limits, suggesting that motion produced by the displacement force was limited by ligamentous restraints only. The clinical assessment of the passive limits of patellar motion should include examination at knee flexion angles of 0 degree and 35 degrees. The manually produced displacement was found to be more reproducible than displacement by the Patella Pusher (P < 0.05). PMID- 8238717 TI - Effects of prophylactic knee bracing on lower extremity joint position and muscle activation during running. AB - The primary purpose of a prophylactic knee brace is to decrease injury potential without compromising joint function; however, the extent that these devices can alter knee muscle function is not clear. This study investigated effects of nonprescription prophylactic knee bracing on lower extremity joint position and muscle activation during running. Six healthy male college-aged recreational runners used a motor-driven treadmill under 6 conditions: fast or slow speeds, with and without wearing a prophylactic knee brace, and with and without wearing a weighted vest. Changes in hip, knee, and ankle joint position and electromyographic activity from 9 lower extremity muscles were monitored during the running cycle weightbearing phase. Results revealed significant changes (P < 0.05) in joint position and electromyographic activity for comparisons of slow versus fast, braced versus nonbraced, and weighted versus nonweighted conditions. From 83% to 89% of the braced and nonbraced comparisons generated significant differences in knee joint position while significant hip and ankle joint position changes occurred in 50% to 58% of comparisons. From 67% to 83% of braced and nonbraced comparisons for the 9 muscles produced significant differences in electromyographic activity. Findings suggest that neuromuscular control is altered when external prophylactic devices are applied. PMID- 8238718 TI - Quadriceps strength and functional capacity after anterior cruciate ligament reconstruction. Patellar tendon autograft versus allograft. AB - Harvesting the central third of the patellar tendon for autograft anterior cruciate ligament reconstruction is thought to compromise quadriceps strength and functional capacity. We compared objective measurements of quadriceps strength and functional capacity in athletes after patellar tendon autograft or allograft anterior cruciate ligament reconstruction. We looked at 33 active male patients (mean age, 24.3 years) who had anterior cruciate ligament reconstructions 12 to 24 months earlier using patellar tendon autograft (N = 15) or allograft (N = 18) techniques. All patients underwent an intensive rehabilitation program. Quadriceps strength and power were assessed by measuring peak torque at 60 and 240 deg/sec, torque acceleration energy at 240 deg/sec, and the quadriceps index using a Cybex II isokinetic testing device. Functional capacity was evaluated based on the results of 3 specially designed functional performance tests and the hop test. Results revealed no significant difference between autograft and allograft groups with respect to any of these parameters. These findings indicate that harvesting the central third of the patellar tendon for autograft anterior cruciate ligament reconstruction does not diminish quadriceps strength or functional capacity in highly active patients who have intensive rehabilitation. Thus, the recommendation to avoid patellar tendon autograft anterior cruciate ligament reconstruction to preserve quadriceps strength and functional capacity may be unnecessary. PMID- 8238719 TI - Quantitative analysis of the measuring capabilities of the KT-1000 knee ligament arthrometer. AB - The KT-1000 arthrometer was evaluated in vitro and in vivo to determine accuracy and quantify effects of potential error sources in clinical application. The KT 1000 arthrometer in vitro accuracy was evaluated by making 30 measurements of 13 known displacements (range, +15 to -15 mm). The effect of applied force on malalignment measurements was evaluated in vitro by making repeated measurements with force applied 5 degrees, 10 degrees, and 15 degrees from the vertical position. The effect of malpositioning the device along the joint line was evaluated in vivo by making repeated measurements 1 cm proximal and 1 cm distal to the joint line. The KT-1000 arthrometer was accurate in vitro (average error, 0.13 mm; SD, 0.12 mm). The range of measurements increased when the angle of force application was increased. Positioning the device 1 cm proximal to the joint line produced larger anterior translation measurements in vivo than those at the joint line (5.8 versus 5.4 mm), while positioning it 1 cm distal produced smaller measurements (4.4 mm). The KT-1000 arthrometer's accuracy indicates great potential for clinical application, but one must ensure that the displacing force is directed properly and the device is positioned accurately over the joint line. PMID- 8238720 TI - Restoration of osseous homeostasis after anterior cruciate ligament reconstruction. PMID- 8238721 TI - Stress fractures of the ulna in tennis players using a double-handed backhand stroke. PMID- 8238722 TI - Refracture of radius and ulna in a female gymnast. A case report. PMID- 8238723 TI - Cyst of the medial meniscus after arthroscopic meniscal repair. PMID- 8238724 TI - Type III-A tibial fracture associated with simultaneous anterior cruciate ligament avulsion from the femoral origin. PMID- 8238725 TI - Meniscal transplantation using fresh and cryopreserved allografts. An experimental study in goats. PMID- 8238726 TI - Comparison of symptomatic versus nonsymptomatic patients with chronic anterior cruciate ligament insufficiency. PMID- 8238727 TI - Migration of a Kirschner wire from the sternum to the right ventricle. A case report. PMID- 8238728 TI - The 'sport' of boxing. PMID- 8238729 TI - Atypical fibroxanthoma. Multiple immunohistologic profiles. AB - The clinical, histologic, and immunohistochemical features of 37 cases of atypical fibroxanthoma (AFX) are presented. Patients ranged in age from 13 to 95 years (mean, 69). Thirty AFXs occurred on the head and neck, and seven lesions developed on the trunk or extremities. The morphologic spectrum varied from a predominant spindle cell pattern with focal cellular pleomorphism to numerous bizarre epithelioid cells with multinucleated giant cells. The spindle cell component in these lesions ranged from 10 to 90% of the constituent cells. Most (31 of 37) AFXs also contained pleomorphic giant cells. Small numbers of S-100 positive dendritic cells were present in 11 cases. Five cases showed variable reactivity with anti-factor-XIIIa. Fifteen (41%) of the AFXs stained for muscle specific actin or smooth muscle actin and 21 (57%) expressed CD68 (detected with monoclonal KP1), a monocyte-macrophage marker. Reactivity for these antigens was seen in all lesional cell types (spindled, epithelioid, and bizarre). Four immunologic profiles were observed: CD68 only (13 cases), actin only (7 cases), double positives (8 cases), and double negatives (9 cases). No significant differences in staining characteristics were observed in the head and neck versus the trunk and extremity lesions. These results expand the immunohistochemical spectrum of AFX, suggest the concept of heterogenous bimodal "fibrohistiocytic" and "myofibroblastic" phenotypes, and provide further evidence that an integrative, nonalgorithmic approach is necessary in the analysis of these and other spindle cell cutaneous lesions. PMID- 8238730 TI - Clinicopathological analysis of 19 cases of isolated carcinoma in situ of the bronchus. AB - Nineteen cases of isolated squamous cell carcinoma in situ (CIS) of the bronchus were described clinicopathologically from among 149 male heavy smokers with roentgenographically occult lung cancer discovered mainly by mass screening performed from 1982 to 1991. All 19 patients had positive sputum cytology tests and negative chest x-ray films and underwent lobectomy (except one who had segmentectomy because of poor lung function). Prior to operation, localization was accomplished by one to eight bronchoscopies using repetitive brush cytology and biopsy. Five cases were bronchoscopically invisible. Polypoid protuberance was noted in three cases, micronodular swelling in three, thickening of spur in five, and mucosal granularity in three. Histology by serial block sectioning showed that there was no nodal involvement in any cases; the maximum length or diameter was 12 mm. Thirteen tumors were < or = 4 mm, four of which were confined to the spur where they occurred. Follow-up data showed a favorable prognosis. Segmentectomy or sleeve resection of bronchus without mediastinal lymph node dissection may be adequate for CIS. PMID- 8238731 TI - Hepatitis B and alterations of the p53 tumor suppressor gene in hepatocellular carcinoma. AB - In areas of the world where hepatitis B and aflatoxin ingestion are common, alterations of the p53 tumor suppressor gene have frequently been reported in hepatocellular carcinoma (HCC). In particular, G-to-T transversions at codon 249 of the p53 gene have been consistently observed in hepatocellular carcinomas in China and sub-Saharan Africa. The goal of this study was to determine the frequency and relationship of p53 gene alterations and hepatitis B in formalin fixed, paraffin-embedded HCCs resected in the United States. Since immunoreactivity for p53 correlates closely with the presence of missense mutations in the p53 gene, we performed immunohistochemical staining with the monoclonal antibody PAb1801. Only seven of 37 cases (19%) demonstrated nuclear accumulation of p53 gene product, in contrast to 10 of 20 cases (50%) of colon carcinoma metastatic to the liver. Staining was not observed in seven liver cell adenomas, 10 cases of focal nodular hyperplasia, or eight cases of cirrhosis. DNA was extracted from formalin-fixed paraffin sections for additional analysis with use of the polymerase chain reaction (PCR). G-to-T transversions of the third nucleotide of codon 249 were demonstrated in only four of 37 cases (11%), three of which had stained with PAb1801. Of 13 patients for whom there was information about a restriction fragment length polymorphism (RFLP) for BstUI within the fourth exon of the p53 gene, allelic loss of p53 was demonstrated in only two cases (15%), both of which stained with PAb1801. Because of previous reports specifically associating hepatitis B with p53 mutations in HCC, we performed nested PCR for hepatitis B virus DNA. Five of 37 cases (14%) contained hepatitis B virus DNA, two of which stained diffusely for p53 and three of which had codon 249 mutations. Our findings indicate that alterations in the p53 gene, particularly at codon 249, are uncommon in HCCs in the United States, and when present are associated with hepatitis B. Since hepatitis B is infrequently associated with HCC in our patient population, the role of p53 alterations in hepatocellular carcinogenesis may not be as significant as in other parts of the world where hepatitis B and aflatoxin are more prevalent. PMID- 8238732 TI - The mechanisms and prognostic significance of seminal vesicle involvement by prostate cancer. AB - To assess the mechanisms and prognostic significance of seminal vesicle involvement (SVI) by prostatic adenocarcinoma, we analyzed 312 radical prostatectomy specimens obtained from patients with T1-T3 prostate cancer. Detailed pathological examination demonstrated three patterns of SVI. Type I involvement was direct spread along the ejaculatory duct complex into the seminal vesicles. Type II involvement was spread outside of the prostate, through the capsule, and then into the seminal vesicle. Type III involvement was characterized by the finding of isolated deposits of cancer in the seminal vesicle with no contiguous primary cancer in the prostate. We found SVI in 64 patients (21%), who have been followed for a mean of 31 months (range 1-101). A defining criterion for progression was clinically apparent local or distant recurrence or a postoperative serum prostate specific antigen (PSA) > or = 0.4 ng/ml (Hybritech). Type I SVI was found in 17 (26%), Type II in 21 (33%), and Type III in 8 (13%) cases. In 18 patients (28%), the pattern of SVI appeared to be a combination of types I and II (categorized as Type I+II). Type III (isolated metastasis) SVI was associated with significantly smaller cancers (median, 3.13 vs. 6.7 cc; p < 0.0005) and fewer positive margins (0 vs. 32%; p = 0.05) than in other types. Type II SVI, with direct extension through the capsule, was associated with a significantly higher risk of lymph node metastasis (8 vs. 33%; p < 0.05). When patients with lymph node metastases were excluded, there was a trend toward a more favorable prognosis (p = 0.09) for patients with type III SVI than with other types. Overall, patients with type III SVI had a progression-free survival rate similar to that of 83 patients with extracapsular extension without SVI. We conclude that the prognostic significance of SVI may not be uniformly ominous; instead, it may depend on the specific mechanism of involvement and the pathologic features of the primary tumor. PMID- 8238733 TI - Epithelial misplacement after biopsy of colorectal adenomas. AB - The question has been raised as to whether the removal of biopsy material from colorectal adenomas can lead to misplacement of tumour fragments into the deeper layers of the bowel wall. We carried out an analysis of histological sections obtained from 21 patients in whom, between 2 and 21 days before removal of their adenoma, forceps biopsy (n = 18), or polypectomy (n = 3) had been performed. The initial histological picture a few days after biopsy is of an ulcer covered by a suppurative exudate and abundant mucus-containing small, free-lying groups of tumour cells. After an average of 7 days, we found misplacement of these groups of cells into the submucosa where, during the course of reparative processes, they became embedded within a capillary-rich granulation tissue. Subsequently, fibrosis of the submucosa develops with persistent mucus pools. Our results thus show that removal of biopsy material from adenomas can result in misplacement of tumour particles into the submucosa. These changes were observed in flat or broad based tumours, and epithelial misplacement appears to be detectable within only a short period after biopsy. For the differential diagnosis, it is important to distinguish these changes from invasive carcinoma. PMID- 8238734 TI - Chemotherapy effects on hepatoblastoma. A histological study. AB - The histopathological features of hepatoblastoma in 17 patients treated with preoperative chemotherapy were compared with those in 11 patients not subjected to chemotherapy during the same 11-year period. Tumor necrosis was more extensive in patients receiving preoperative chemotherapy. Two tumors, however, were apparently unaffected by chemotherapy. There was no obvious correlation between the extent of necrosis and the number of courses of chemotherapy. There also seems to be no evidence of preferential ablation of a particular morphological type of tumor. The most notable feature in cases treated with chemotherapy was the extensive presence of osteoid. Osteoid was present in 36% of untreated cases, occupying < 5% of the surface area, compared with 82% in the treated group. In seven cases, osteoid occupied > 40% of the surface area. This finding raises speculation about the role of chemotherapy in the maturation of tumors that have an inherent ability to differentiate. A long-term study is needed to clarify the prognostic significance of mature heterologous elements in hepatoblastoma. PMID- 8238735 TI - Hepatic sarcoidosis. Clinicopathologic features in 100 patients. AB - The patterns of hepatic injury were studied in 100 patients with a diagnosis of sarcoidosis and clinical evidence of liver disease that led to diagnostic liver biopsy. Granulomas were present in all patients; they occupied from < 1% to > 90% of the total volume of tissue examined and were most often located in the portal/periportal region. In none of the 100 cases were infectious organisms identified by special stains, culture, or serology. In 99% of cases, these granulomas were noncaseating; in one of the 100 cases central caseation was noted. In addition to the granulomas present in all biopsies, three broad categories of histologic change were found: cholestatic (58%), necroinflammatory (41%), and vascular (20%). Among those with cholestasis, 19 patients had bile duct lesions similar to primary biliary cirrhosis, whereas another 13 had a pattern of periductal fibrosis reminiscent of primary sclerosing cholangitis. In 37 patients with chronic cholestasis, a decrease in the number of bile ducts (ductopenia) was noted. Twelve patients had an acute cholangitis suggestive of mechanical obstruction--although no clinical evidence of ductal obstruction was found. Necroinflammatory changes included spotty necrosis suggesting hepatitis of diverse etiologies (including viral infection and drug reaction) and chronic portal inflammation suggestive of chronic active hepatitis. Vascular changes consisted of sinusoidal dilatation (14 cases) and nodular regenerative hyperplasia (9 cases). In 6% of the patients, the only changes in the biopsy were those of granulomatous inflammation; each of these patients had a dominant mass ("sarcoidoma"), which had been biopsied to rule out tumor. Fibrosis was seen in 21% of the biopsies--periportal (13%), bridging (2%), or cirrhosis (6%). It is clear that sarcoidosis can cause progressive liver disease with a wide array of histologic features that can mimic those of other primary liver diseases. PMID- 8238736 TI - Diagnostic value of AgNOR staining in follicular cell neoplasms of the thyroid: comparison of evaluation methods and nucleolar features. AB - The diagnostic value of argyrophil staining of nucleolar organizer regions (AgNOR) was studied in 95 nonneoplastic and neoplastic follicular lesions of the thyroid. Different AgNOR parameters such as number, size, and distribution pattern were determined using digital image analysis. In addition, nuclear and nucleolar size as well as the percentage of nucleoli touching the nuclear membrane (nucleolar margination) were assessed. A stepwise increase in nuclear size and AgNOR counts from normal thyroid tissue to follicular adenoma as well as from differentiated follicular to anaplastic carcinoma was found (mean nuclear area [micron2]/mean AgNOR number per cell: 21.5/1.6 vs. 34.4/3.5 and 45.3/5.0 vs. 66.5/10.8, p < 0.01/p < 0.001). There was, however, no clear separation between these diagnostic groups. In contrast, an almost total discrimination between follicular adenoma and carcinoma was achieved by quantification of AgNORs per tumor cell nucleolus (AgNOR distribution score). In benign adenomas, 3.3% (range, 0-8.8%) of the cells showed nucleoli with at least five AgNOR dots within one focal plane, whereas in follicular carcinomas, the corresponding value was 34.1% (range, 12-75%). Two of four cases of so-called atypical adenomas showed values in the range of benign adenomas, and two were in the range of follicular carcinomas. In comparison with other nuclear and nucleolar parameters, the AgNOR distribution score proved the most valuable diagnostic criterion for the cytomorphological differentiation between follicular adenoma and carcinoma of the thyroid. PMID- 8238737 TI - Aggressive natural killer cell lymphoma/leukemia. A recently recognized clinicopathologic entity. AB - We report a comprehensive study of a case of aggressive natural killer cell lymphoma/leukemia, which is characterized by young male predominance, rapidly progressive clinical course, and presence of lymphadenopathy, hepatosplenomegaly, and bone marrow involvement. The leukemic phase is frequently preceded by pancytopenia. The diagnostic clues are the detection of cytoplasmic granules in tumor cells on Wright-Giemsa-stained tissue imprints or smears and a selective loss of T-cell antigens. Immunophenotyping is decisive in making the final diagnosis by showing positive natural killer cell markers (CD16, CD56, and/or CD57), CD2, CD11c, and Ia, but negative CD3, T-cell receptor heterodimers, terminal deoxynucleotidyl transferase, and B-cell markers. Genotyping always shows germline configuration in both immunoglobulin and T-cell receptor genes. The unique feature in this case is its presentation as a testicular lymphoma, which has not been previously reported. Polymerase chain reaction was performed in this case but failed to detect human T-cell leukemia virus type I/II provirus. It is important to recognize this new entity as it is a highly aggressive disease with a rapidly progressive clinical course and fails to respond to any chemotherapeutic regimen available. PMID- 8238738 TI - Program and abstracts of the Joint Annual Meeting of the American Society of Tropical Medicine and Hygiene and the American Society of Parasitologists. Atlanta, Georgia, October 31-November 4, 1993. Abstracts. PMID- 8238739 TI - Festschrift issue devoted to John S. Najarian, in honor of the 25th anniversary of his becoming chairman of the surgery department at the University of Minnesota. PMID- 8238740 TI - The Minnesota Program under John S. Najarian. PMID- 8238741 TI - Use of novel therapeutic agents for the treatment of serious infection. AB - The use of biologic reagents directed against invading microbes and deleterious aspects of the host response is presently under intense investigation. The development and use of these reagents have contributed significantly to our understanding of the host response to infection, and they probably represent the first true foray into the area of immunotherapeutic manipulation. Thus, the effects of the administration of anti-endotoxin and antitumor necrosis factor alpha monoclonal antibodies and the interleukin-1 receptor antagonist are being examined in large-scale clinical trials, and tumor necrosis factor-binding protein and bactericidal/permeability-increasing protein also may undergo clinical testing. Much controversy has already arisen regarding the manner in which these therapeutic reagents will be best utilized. The enormous amount of information that has already been obtained concerning the use of these reagents assuredly will contribute significantly to our understanding of the diagnosis and treatment of serious infection and will set the stage for the development and appropriate testing of additional new diagnostic and therapeutic reagents during the current decade. PMID- 8238742 TI - Evolution of kidney, pancreas, and islet transplantation for patients with diabetes at the University of Minnesota. AB - Transplantation began at the University of Minnesota in 1963. Treatment of diabetes and its complications has been emphasized since 1966, when the first pancreas-kidney transplant was done. Of 3,640 kidneys transplanted by 1992, 1,373 were for diabetic recipients, including 658 from living donors and 715 from cadaver donors. The results progressively improved; since 1984, survival rates of kidney grafts have been similar for diabetic and nondiabetic recipients, with three fourths of the grafts functioning at 4 years. As of 1992, 501 pancreas transplants had been done, including 170 simultaneous with a kidney, 142 after a kidney, and 188 alone for nonuremic diabetic patients; again, the results have improved: by the 1990s, graft survival rates were similar in the 3 recipient categories. Successful pancreas transplants have been shown by our coworkers to stabilize or improve neuropathy and prevent recurrence of diabetic nephropathy in kidney grafts. In an attempt to simplify endocrine replacement therapy, we have done 63 human islet transplants, 34 as allografts for patients with type I diabetes and 29 as autografts after total pancreatectomy to treat chronic pancreatitis. Insulin independence occurs for about 50% of islet autograft recipients. Two recent islet allograft recipients treated with 15-deoxyspergualin have had sustained insulin independence. We anticipate that endocrine replacement therapy by transplantation will become routine for diabetic patients as methods to prevent rejection are refined. PMID- 8238743 TI - Lumenal route for intestinal radioprotection. AB - The intestine, being highly sensitive to radiation injury, is the limiting factor in the delivery of abdominal and pelvic radiation. Attempts at intestinal radioprotection have generally involved systemic administration of agents that are moderately toxic and that may protect the target malignancy. The lumenal route for intestinal radioprotection provides a high concentration at the mucosa without significant systemic effects. This paper reviews studies of intestinal radiation radioprotection by intralumenal agents. Our model involved irradiation of exteriorized rat small bowel containing drugs in isolated segments. Surviving crypts were quantitated as a measure of injury 5 days later, and the individual segments were compared. Modest protection was found with lumenal alkalinization and with neutralization of bile salts or pancreatic enzymes. Similar protection was identified with some nonsteroidal anti-inflammatory agents, methylprednisolone, and selected antioxidants. Marked protection was observed with lumenal Lazaroid. PMID- 8238744 TI - Controversies in primary breast cancer management. AB - Adequate locoregional treatment of patients with primary operable breast cancer involves the control of multicentric disease in the breast and axillary dissection to stage the disease and control it in the axilla, when present. Two options, having equal survival rates in prospective, randomized studies, are breast preservation and mastectomy. In breast preservation, adequate tumor excision with clear histologic margins and axillary dissection is followed by breast irradiation. The mastectomy option involves no radiotherapy and can be followed by reconstruction. Careful selection of patients and a detailed description of the pros and cons of each approach should be undertaken on an individual basis. PMID- 8238745 TI - Liver transplantation and the Najarian footprint. PMID- 8238746 TI - Evolution of the bioartificial liver: the need for randomized clinical trials. AB - The pursuit of a bioartificial liver is well documented in the literature. Early techniques of artificial liver support that have undergone clinical testing included simple exchange transfusions, extracorporeal xenogeneic or allogeneic liver perfusion, cross-circulation, hemodialysis, charcoal hemoperfusion, and plasmapheresis with plasma exchange. These techniques failed because they were unable to adequately support those hepatic functions essential for survival and because they lacked a back-up therapy, such as liver transplantation, for irreversible forms of liver disease. The concept evolved that hepatic functions essential for survival would be best performed by hepatocytes in an apparatus that allowed sustained or repetitive application. The best results have been achieved with bioartificial liver technologies that employ hepatocytes as implantable systems or extracorporeal devices. Implantable bioartificial liver systems include hepatocytes that have been on coated microcarrier beads, within microencapsulated gel droplets, within biodegradable polymeric substrates, or as spheroid hepatocyte aggregates. Extracorporeal systems include hepatocytes in suspension, on flat plates, and in hollow fiber bioreactors. Several extracorporeal systems have undergone extensive animal testing and are entering the early stages of human clinical trials. Randomized trials are needed to establish the value of bioartificial liver support in the treatment of patients with acute hepatic failure or as a bridge to liver transplantation. PMID- 8238747 TI - Past, present, and future for surgical needles and needle holders. AB - During the last two decades, major advances in surgical needle and needle holder technology have markedly improved surgical wound repair. These advances include quantitative tests for surgical needle and needle holders performance, high nickel maraging stainless steels, compound curved needles, needle sharpening methods, laser-drilled holes for swages, needle:suture ratios of 1:1, and the atraumatic needle holder. PMID- 8238748 TI - Hemodynamic, respiratory, and metabolic effects of laparoscopic cholecystectomy. AB - In 10 patients undergoing laparoscopic cholecystectomy, creation of pneumoperitoneum caused immediate venous hypertension and stasis in the lower extremities as measured by percutaneous catheter and duplex scanning. These changes disappeared after deflation. As measured by spirometry, significant reductions in forced vital capacity of 23% and forced expiratory volume in 1 second of 22% were present 24 hours after surgery, and plasma interleukin-6 levels rose to 18 pg/mL. The visual analogue scale of resting pain increased to a median value of 2.5 postoperatively. When compared with other studies of open cholecystectomy, our results showed fewer restrictions of ventilation, lower cytokine levels, and lower pain scores. The minimal soft tissue trauma and early ambulation after laparoscopic cholecystectomy may decrease the risk of thrombosis despite an acute episode of venous stasis. PMID- 8238749 TI - Pancreatic islet transplantation: is purification necessary? AB - Successful islet allotransplantation in diabetic patients has been slow to develop. In the past, it has been assumed that the low success rates are, in part, due to difficulty in obtaining high-quality human islet preparations. The assumption has been that purified islets are essential because exocrine contamination hampers engraftment and makes islets more vulnerable to rejection by increasing graft immunogenicity. We reviewed the existing experimental data on the consequences of exocrine contamination and concluded that most clinical trials of islet allotransplantation have involved efforts to over-purify the allograft. Until antigen-specific tolerance or the biohybrid artificial pancreas is a reality, the best strategy for clinical trials is to test new immunosuppressive protocols in a search for a more effective way of modifying the host's immune response while maximizing islet yield by using less pure preparations. PMID- 8238750 TI - Aggressive metastasectomy for pulmonic sarcomatous metastases: a follow-up study. AB - Pulmonary metastases are the primary cause of death due to bone and soft tissue sarcomas. We have previously shown that an aggressive approach and a new technique of multiple pulmonary metastasectomies have resulted in improved survival for patients with pulmonary metastases. In this follow-up study, an expanded database of patients was retrospectively analyzed to determine survivability as well as to evaluate potential prognostic indicators. Forty-nine patients, 26 of whom had osteogenic sarcoma (OGS), were evaluated. A number of patients had been referred from other institutions where their disease had been considered inoperable because it was extensive or recurrent. Using lateral thoracotomies exclusively, employment of a laser technique, and excision of minimal pulmonary parenchymal tissue, we performed aggressive metastasectomy. A mean of 3.0 thoracotomies was performed, in which an average of 10.2 nodules per thoracotomy were excised. Operative morbidity and mortality were minimal. The disease-free interval, the number of nodules resected, the number of thoracotomies performed, and the size of the nodules were evaluated as potential prognostic indicators. Statistically significant correlation could be established only for the size of the nodules resected. The 5-year survival rate for all patients was 39%; it was 24% for patients with OGS and 71% for those without OGS. Aggressive surgical resection of pulmonary metastases from bone and soft tissue sarcoma should be considered when there is control of local disease, no evidence of extrapulmonary metastasis, and adequate post-resection pulmonary reserve. The presence of bilateral, extensive, or recurrent disease is not a contraindication to thoracotomy. Aggressive resection of multiple nodules and improved chemotherapy appear to prolong survival of these patients when compared with survival rates of historical control subjects. PMID- 8238751 TI - Fontan's procedure in two stages. AB - Since 1985, 381 patients with various cardiac malformations that share the central common feature of only one effective ventricle have been treated with some modification of Fontan's operation. Since 1989, the Fontan operation has been staged by associating the superior vena cava with the branch pulmonary arteries (hemi-Fontan) initially followed, some months later, by associating the inferior vena cava to the branch pulmonary arteries (completion Fontan). The difference in early mortality for a primary-Fontan operation (16%) compared with a completion-Fontan operation (7%) is substantial (p < 0.05). Since January 1991, mortality associated with the hemi-Fontan operation has been 5.5% (7 deaths in 127 patients). A systematic staged approach to Fontan's operation has been undertaken with a hemi-Fontan operation in patients who are 6 months of age and a completion Fontan operation when those patients are 12 to 18 months of age in an effort to reduce the volume load of the ventricle as early as possible, to minimize intermediate mortality from the palliated state, and to reduce the impact of rapid changes in ventricular geometry and diastolic function that can accompany either the hemi-Fontan or primary Fontan operation, but that are lethal only after a primary Fontan operation. PMID- 8238752 TI - Common pathway of endothelial-leukocyte interaction in shock, ischemia, and reperfusion. AB - Impaired blood flow in shock and ischemia results in significant organ dysfunction and failure of critical cellular functions. Although some cellular function can proceed via anaerobic mechanisms, a point is reached at which restoration of blood flow and oxygen delivery does not result in restoration of function ("refractory shock" or the "no-reflow phenomenon"). But even if blood flow is restored after shock or ischemia, a second mechanism of cellular injury is initiated: monocytes and neutrophils are activated, resulting in an inflammatory response. Current evidence suggests that the activation of inflammatory cells is triggered by substances from ischemic or injured vascular endothelium that cause leukocyte adherence, activation, and further injury. This review summarizes the current literature on endothelial cell, monocyte, and neutrophil interactions in reperfusion injury after shock or ischemia and suggests how a recently described peptide from the vascular endothelium may play an important role in the cascade. PMID- 8238753 TI - Surgical hazards posed by marine and freshwater animals in Florida. AB - Marine and freshwater animals can cause injury to humans by biting, stinging, being poisonous to eat, and causing infections. Biting aquatic animals in Florida include sharks, barracudas, alligators, and moray eels. Devitalized tissue should be debrided, and vascular, neurologic, and tendinous injuries should be repaired. Radiographs should be obtained to examine the injury sit for fractures and retained foreign bodies (teeth). The spines of stingrays and marine catfish can cause soft tissue injury and infection. The spine has a recurved, serrated shape that may cause further injury and break if it is pulled out. The venom may cause local tissue necrosis requiring debridement. Soft tissue infections with marine Vibrio bacteria can occur after eating raw oysters or receiving even minor injuries from marine animals. Thirty-one individuals developed soft tissue infections, 49 developed sepsis, and 23 developed both sepsis and soft tissue infection with marine Vibrio species during a 12-year period. Sixteen patients developed necrotizing soft tissue infections. Treatment is with antibiotics and debridement when necrosis occurs. PMID- 8238754 TI - Altemeier tumors? PMID- 8238755 TI - Laparoscopy and gastrointestinal cancer. PMID- 8238756 TI - Intranasal corticosteroids and cromolyn. PMID- 8238757 TI - Salvage chemotherapy in recurrent head and neck cancer: the Institut Gustave Roussy experience. PMID- 8238758 TI - Real-time ultrasonography in the otolaryngology office setting. PMID- 8238759 TI - Computer-quantified positional nystagmus in normals. AB - INTRODUCTION: Most current authorities agree that, in the absence of visual fixation, low intensity nystagmus can be recorded by electronystagmography (ENG) in clinically normal people. However, there are some notable dissenters, and the reported incidence, intensity, and direction of "normal" spontaneous/positional (S/P) nystagmus vary widely across laboratories. The highly subjective element introduced by manual interpretation of position test records compounds the problem of characterizing "normal" S/P nystagmus. MATERIALS AND METHODS: A computerized ENG analysis system which removes this element of subjectivity is described, and results of a study validating its results against those of experienced ENG interpreters are reported. RESULTS: The computer found an S/P nystagmus behind closed eyelids in at least one of five test positions in 42 of 51 clinically normal subjects (82.3%). This "normal" S/P nystagmus had a statistically significant overall directional bias toward left beating, but was affected by position--tending toward right beating in the left lateral position. The upper 95.4% confidence limits for computer-measured slow phase velocities of "normal" S/P nystagmus were -5.5 degrees/sec (left beating) and +4.4 degrees/sec (right beating). CONCLUSION: Possible explanations for "normal" S/P nystagmus include clinically undetected pathology, normal asymmetries in the vestibulo ocular reflex, and an imprinted oculomotor pattern such as might be created by reading. PMID- 8238760 TI - Hypercoagulability in otologic patients. AB - INTRODUCTION: Three patients with otologic disorders developed complications related to hypercoagulability. This report was prepared to increase the awareness among otolaryngologists of the diagnosis and management of hypercoagulable states, and to encourage expert consultation when indicated. MATERIALS AND METHODS: One patient with dural venous sinus thrombosis complicating an otitis media, one patient with a large glomus jugulare tumor, and one patient with a sudden sensorineural hearing loss were treated by the otolaryngology service. Suspicious thromboembolic events were evaluated by the hematology service, and appropriate anticoagulative therapy was recommended. RESULTS: One patient with an inherited hypercoagulopathy and two patients with acquired hypercoagulopathies were treated for otologic problems. Two of the patients were stabilized and discharged on life-long anticoagulation therapy. The third patient, in spite of intensive medical and surgical support, eventually succumbed to complications to which an acquired hypercoagulable state made a significant contribution. Expert consultants made the hematologic diagnoses and treatment recommendations. CONCLUSION: Otolaryngologists should be aware that hypercoagulable states may now be more accurately diagnosed and characterized, and that thorough investigation of thromboembolic events may affect treatment decisions. PMID- 8238761 TI - Carcinoma ex oncocytic Schneiderian (cylindrical cell) papilloma. AB - INTRODUCTION: This study reviews the clinicopathologic features of carcinoma presenting in oncocytic Schneiderian papilloma (OSP; cylindrical cell papilloma), a rare histologically distinctive papilloma sharing features with inverted papilloma. MATERIALS AND METHODS: Nine cases of carcinoma ex OSP (1970-1990), six from the files of the Armed Forces Institute of Pathology (Washington, DC) and three from the surgical pathology files, Presbyterian-University Hospital and Eye and Ear Institute (Pittsburgh, PA), were studied. One case was previously reported. Clinical data were obtained from the patients' medical records. All available histologic slides were reviewed. RESULTS: The patients (eight men, one woman) ranged in age from 48 to 83 years (mean age, 74 years). The carcinoma and OSP were synchronous in eight patients. In one case, the carcinoma occurred 8 years after OSP was diagnosed on biopsy. The presenting manifestations included nasal obstruction, "polyps," epistaxis, and proptosis. Radiographically, a sinonasal mass was present, often with extension to adjacent sites and bone destruction. Most patients received postoperative irradiation. Of six patients with follow-up, four died of carcinoma and two were alive without disease at 1.5 and 7 years, respectively. The types of carcinoma were: squamous cell (n = 6), high-grade mucoepidermoid (n = 2), and sinonasal undifferentiated (n = 1). Histologic transition was observed between carcinoma and dysplastic/benign OSP in eight cases. CONCLUSION: Demonstration of histologic continuity between carcinoma and dysplastic OSP epithelium in eight of our nine cases indicates origin of carcinoma from the OSP, rather than a coincidence of the two neoplasms. Although histologically benign, OSP should be totally excised at diagnosis because of a potential for malignant transformation. This is a US government work. There are no restrictions on its use. PMID- 8238762 TI - Reconstruction of the anterior commissure with a free mucosal flap. AB - INTRODUCTION: To prevent stenosis and adhesion of the anterior commissure in the laryngeal reconstruction, we devised a technique of relining using a free labial mucosal graft and human fibrin glue. MATERIALS AND METHODS: A single large piece of mucosal flap was prepared, folded at the anterior commissure, and sutured onto the raw wound of the larynx from both sides of the posterior glottis. Alternate suturing from both right and left sides was performed anteriorly, and the wound on the laryngeal lumen was fully covered by the mucosal graft. We applied this technique on a patient with laryngeal stenosis and two patients with laryngeal cancer. RESULTS: Without the use of a stent, we successfully reconstructed the anterior commissure and obtained a good result in voice quality in each case. CONCLUSION: Large mucosal defects including the anterior commissure should be relined to acquire good voice quality. Relining using a free mucosal flap was a useful method since there were no limitations to size and placement. The adhesive property of human fibrin glue was indispensable in this procedure. PMID- 8238763 TI - Head and neck cellulitis in hospitalized adults. AB - INTRODUCTION: Our purpose was to review the clinical features of head and neck cellulitis in hospitalized patients. MATERIAL AND METHODS: A retrospective review was undertaken of 147 cases of head and neck cellulitis seen over a 15-year period at a community Hawaiian hospital. RESULTS: Otologic cellulitis was mainly related to otitis externa, caused by Pseudomonas aeruginosa, although some cases had infected pierced ears or ear cysts. Cervical cellulitis was associated with malignancy, dental infection, or lymphadenitis; these patients often had positive blood cultures. Facial cellulitis was almost always caused by Staphylococci and Streptococci, usually preceded by dental infections, traumatic abscesses, or sinusitis. All study patients recovered completely, except for one patient who developed cavernous vein thrombosis and oculomotor palsy. Seventy-five percent of the patients recovered with parenteral antibiotics alone; the other patients required abscess drainage and/or other surgical procedures, especially if neck infection was present. CONCLUSION: Although head and neck cellulitis cases have grave potential consequences, most patients do very well with proper antibiotics and appropriate surgical drainage. PMID- 8238764 TI - Spindle cell carcinoma of the larynx with simultaneous carcinoma of the thyroid gland. PMID- 8238765 TI - Retropharyngoesophageal abscess associated with a swallowed bone. PMID- 8238766 TI - Sclerosing cervicitis: an unusual cause of syncope. PMID- 8238767 TI - Labyrinthitis ossificans. PMID- 8238768 TI - Simultaneous functional laryngeal stridor and functional aphonia in an adolescent. PMID- 8238769 TI - Meta-analysis of the carotid artery. PMID- 8238770 TI - Death in Anchorage in 1988 and summary of deaths 1985-1988. AB - In 1988, 832 persons died in Anchorage, Alaska. Twenty-seven percent succumbed each to cardiovascular disease and cancer, 20 percent to violence including alcoholism, and the remaining 26 percent to other disorders. These proportions were about the same as in the four-year period 1985-88. Violence continued to account for more loss of life in Anchorage than nationally. PMID- 8238771 TI - Quantitative observations of hymens in prepubescent females selected for non abuse. AB - The maximum hymenal opening was evaluated quantitatively in 111 prepubescent females during routine physical examinations in a pediatric subspecialty office. For comparison, an additional 53 females referred by child protective agencies were also examined. Analysis of data show "non-abused" groups may be separated statistically from "abused" groups on the basis of area of hymenal opening. The mean area for the "non-abused, non-masturbate" group was 6.4 mm2. The upper limit of area (mean + 3 S.D.) of hymenal opening in this group was 24.1 mm2. A child having a hymenal opening diameter of 6.94 mm or less has a 99% chance of being in the "non-abused" group. The area of hymenal opening for "non-abused" groups did not change with increasing age, height or weight. A skilled pediatrician knowledgeable in the area of sexual abuse may obtain clinically relevant information with ordinary office equipment and trained personnel. Regular and repeated observations of genitalia during routine health maintenance examinations are vital baseline measurements for the physical and mental health of young female patients. PMID- 8238772 TI - Homicide in Alaska Natives. PMID- 8238773 TI - Pediatric lead level screening. PMID- 8238774 TI - Alaska's own family practice residency program. PMID- 8238775 TI - Alaska Fetal Alcohol Syndrome Prevention Project. PMID- 8238776 TI - Medical practice in western Alaska around 1900. 1939. PMID- 8238777 TI - A look at hospital care in New Archangel (1835-1840). PMID- 8238778 TI - Distinction between diffuse cutaneous malignant follicular center cell lymphoma and lymphoid hyperplasia by computerized nuclear image analysis. AB - The difficult differential diagnosis between the diffuse variants of cutaneous lymphoid hyperplasia (CLH; synonym; pseudolymphoma) and malignant follicular center cell lymphomas (FCCL) often requires a multidisciplinary approach. Eighteen CLH and 11 FCCL, diagnosed by conventional histology and immunophenotyping and subsequently examined with a polymerase chain reaction to show clonal immunoglobulin heavy-chain gene rearrangements, were subjected to a novel type of automated nuclear image analysis. Of all nuclear parameters tested in azure A-stained semithin sections, the mean nuclear profile area (TN) of lymphoid cells was the best criterion to distinguish between CLH and FCCL (p = 9 x 10(-6)). Additional distinctive features, in the order of decreasing significance, were the SD of TN; all chromatin textural parameters combined; and the light and the dark fractions of the central nuclear profile areas. Parameters related to the chromatin pattern were independent of nuclear profile size in FCCL, but not in CLH. Two lesions registered as CLH displayed the nuclear characteristics favoring this diagnosis, but showed B-cell monoclonality at the DNA level. In conclusion, computerized nuclear image analysis is a helpful additional diagnostic tool in the evaluation of diffuse CLH and cutaneous FCCL. PMID- 8238779 TI - Proliferating trichilemmal cyst. Report of four cases, two with nondiploid DNA content and increased proliferation index. AB - Proliferating trichilemmal cyst (PTC) is an uncommon tumor that usually arises on the scalp of elderly women. Its biologic nature is that of a benign lesion with occasional local recurrences. PTC can be confused both grossly and microscopically with squamous cell carcinoma and malignant PTC. Distinction between these lesions has historically been made on a histologic basis. We present four cases of PTC. Histologically, the lesions consisted of lobules of basaloid cells admixed with larger pale-staining cells with abrupt trichilemmal type of keratinization and peripheral palisading embedded in a fibrous stroma. Flow cytometry performed on nuclear extracts of the lesions revealed two of the four lesions to have nondiploid DNA content. Proliferation index, measured by immunohistochemical staining with Ki-67 monoclonal antibody along with mitotic rate count, was higher in the two nondiploid lesions as compared with their diploid counterparts. The results raise the question of the significance of aneuploidy and increased proliferation indices in otherwise benign PTC. PMID- 8238780 TI - Aggressive angiomyxoma of the pelvioperineal region. Immunohistological and ultrastructural study of seven cases. AB - We describe seven cases of aggressive angiomyxoma of the pelvioperineal region. In three cases, we observed immunohistochemical signs of myoid differentiation in the tumor cells. In one of these cases, we could confirm the myoid differentiation at the ultrastructural level. The other four cases were devoid of this differentiation. We further discuss the relationship of aggressive angiomyxoma to the recently described angiomyofibroblastoma of the vulva. PMID- 8238781 TI - Monoclonal antibody Ber EP4 distinguishes basal-cell carcinoma from squamous-cell carcinoma of the skin. AB - Twenty-two cases of basal-cell carcinoma (BCC) and 21 cases of squamous-cell carcinoma of the skin (SCC) were stained for Ber EP4 using indirect immunofluorescence and avidin-biotin-peroxidase techniques. The results showed that all BCCs were diffusely and intensely labeled, whereas none of the SCCs expressed Ber EP4, irrespective of their histological type or grade of differentiation. This difference may prove useful in (a) histologically differentiating between these two neoplasms; and (b) in clarifying the histogenesis of neoplasms considered to be intermediate between BCC and SCC, or derived from BCC. In addition, because significant differences between the staining of hair follicles and BCC were noted, evaluation for Ber EP4 expression could assist in distinguishing between BCC and hair follicles in Mohs micrographic sections. PMID- 8238782 TI - Late infantile ceroid-lipofuscinoses. An ultrastructural study. AB - The aim of the present study is to investigate further the ultrastructural skin patterns in five cases of late infantile ceroid-lipofuscinosis: two of these were classic forms, the others were variants. Ultrastructural examinations of skin biopsies revealed the presence of characteristic cytosomes; typical lipofuscin, consisting of osmiophilic granular materials, curvilinear bodies, and fingerprint bodies. Different ultrastructural profiles were found simultaneously in each case, without a significant prevalence of any specific one, and were often associated with lipid droplets. These inclusions were found in several epidermal and dermal cells. A different degree of involvement of the myelinated sheaths in the five cases was observed. This difference could be genetically determined or perhaps related to different stages of the disease. The findings demonstrate the involvement of clinically unaffected skin and confirm the relevance of electron microscopic studies in diagnosing these disorders. In fact, recognition of typical ultrastructural changes is a valuable diagnostic tool that can be used in supplementing clinical and electrophysiological examinations, especially when the metabolic error is unknown and no diagnostic biochemical assay is available. PMID- 8238783 TI - Thickness is not an accurate gauge of prognosis of primary cutaneous melanoma. PMID- 8238784 TI - Lesion thickness and prognosis in melanoma: horses are not zebras. A response to Green and Ackerman. PMID- 8238785 TI - Melanoma thickness and prognosis. PMID- 8238786 TI - Do not abandon the Breslow thickness. PMID- 8238787 TI - Is tubular apocrine adenoma a distinct clinical entity? AB - A 75-year-old Japanese man developed a tubular apocrine adenoma (TAA) (tubulopapillary hidradenoma with apocrine differentiation, a rare skin tumor), within a long-standing organoid nevus on the parietal area of his scalp. Histologically, the tumor consisted of dilated ductlike areas with some atypism and apocrine glandlike areas surrounded by myoepithelial cells. The superficial part of the tumor was connected to the epidermis and showed some of the characteristics of syringocystadenoma papilliferum (SCAP). The close relationship between TAA, SCAP, and papillary eccrine adenoma (PEA) is discussed. According to 19 reported cases of TAA, SCAP might occur together with TAA when they are preceded by an organoid nevus, and they might represent a spectrum of disease. Although TAA and PEA may represent another spectrum (designated as tubulopapillary hidradenoma), the relationship to SCAP should be considered in understanding and diagnosing an intermediate case. PMID- 8238788 TI - Cutaneous ganglion cell tumor of the skin. Case report and review of the literature. AB - We describe the histological and immunological findings of a cutaneous tumor composed of mature ganglion cells. The tumor was ganglioneuromatous but differed from other ganglioneuromas in that the neurofibromatous proliferating components were absent. PMID- 8238789 TI - Giant trichoblastoma on the scalp. AB - We describe a case of giant trichoblastoma that had been present for many years on the scalp of a 69-year-old man. The specimen was examined histopathologically, immunohistochemically, and ultrastructurally. Histopathologic examination showed that neoplastic cells were arranged in different patterns from area to area, including rippled, adenoid, and palisading patterns. In some areas of the neoplasm, foci of sebaceous differentiation were also present. The ultrastructural characteristics of neoplastic cells support the notion that trichoblastomas are neoplasms composed mostly of follicular germinative cells. PMID- 8238790 TI - Syringomatous carcinoma of the scalp presenting as a slowly enlarging patch of alopecia. AB - A case of well-differentiated syringomatous carcinoma in a 50-year-old woman is reported. The tumor was located on the scalp and manifested as a slowly enlarging patch of alopecia. It was misdiagnosed as alopecia areata for years and subsequently was erroneously considered to be a benign syringoma on the basis of the results of a punch biopsy. When the correct diagnosis was made, the patient was referred to a plastic surgery department for the complete removal of her lesion. PMID- 8238791 TI - Should Sutton nevus really be called Grunewald-Sutton nevus? PMID- 8238792 TI - A critical analysis of textbooks of dermatopathology in historical perspective. Part 9. PMID- 8238793 TI - Pityrosporum orbiculare and atopic eczema. PMID- 8238794 TI - Allergens of Pityrosporum ovale and Candida albicans. I. Cross-reactivity of IgE binding components. AB - In atopic dermatitis (AD), a high prevalence has been reported of type I reactions and specific IgE to extracts of the commensal lipophilic skin yeast Pityrosporum ovale. In the present study, a highly significant correlation (r = 0.77) was found between levels of anti-P. ovale IgE and of IgE reacting with extracts of Candida albicans, both measured by a sensitive ELISA method. In a series of 128 AD sera, 34 sera reacted positively with both yeast extracts, 38 reacted with P. ovale but not with C. albicans, and only one of the 56 anti-P. ovale-negative sera showed a very weak reaction with C. albicans. The correlation was due to a marked cross-reactivity, as shown by inhibition ELISA. Fluid-phase preincubation of double-positive sera with either of the two yeast extracts resulted in a dose-dependent, and at high concentrations complete, inhibition of the IgE reactions with both coated P. ovale and C. albicans allergens. Mutual inhibition of IgE-binding could also be achieved with pools of glycoproteins and/or polysaccharides isolated from the crude extracts by Con A affinity chromatography. P. ovale allergens were, however, more potent fluid-phase inhibitors than the corresponding C. albicans components. The apparently higher avidity for P. ovale allergens suggests that these antiyeast IgE antibodies in AD result from sensitization to P. ovale and cross-react with C. albicans. PMID- 8238795 TI - Allergens of Pityrosporum ovale and Candida albicans. II. Physicochemical characterization. AB - Pityrosporum ovale has recently been recognized as a source of allergens to which many patients with atopic dermatitis (AD) show type I skin reactions and specific IgE antibodies. In this study the IgE-binding components and/or epitopes in P. ovale extract were shown to be partially sensitive to pronase or trypsin treatment, whereas periodate oxidation resulted in a complete loss of IgE-binding capacity, thus suggesting the involvement of carbohydrate structures. In Con A affinity chromatography most of the IgE-binding capacity of crude P. ovale extract bound to the column, and could be eluted with mannoside. Gel filtration on Sephacryl S-400 revealed a marked heterogeneity with respect to molecular mass, with most of the IgE-binding activity associated with high-mol.-mass fractions (from 5 x 10(4) up to 2 x 10(6) Da). A similar heterogeneity was found after chromatofocusing, with IgE-binding in the whole pI-range from 7.0 to 4.0. Essentially identical results were obtained with extracts of Candida albicans, in agreement with the previously shown cross-reactivity of IgE-binding components in the two yeast extracts. In inhibition ELISA, gel filtration and chromatofocusing fractions containing components with widely different mol. mass or pI showed complete reciprocal cross-inhibition, and were all capable of inhibiting the binding of IgE to unfractionated extracts. We therefore conclude that the cross reacting anti-P. ovale/anti-C. albicans IgE antibodies in the sera of AD patients are mainly directed at a restricted number of carbohydrate epitopes that are expressed on a heterodisperse range of high-mol.-mass components, probably mannans or mannoproteins. PMID- 8238796 TI - Neutrophil chemotactic activity (NCA) in nasal secretions from atopic and nonatopic subjects. Effect of antigen challenge. AB - In order to elucidate the mechanism responsible for infiltration of nasal mucosa by granulocytes, we tested neutrophil chemotactic activity (NCA) in nasal lavages, by the modified Boyden chamber method, in 16 patients with perennial allergic rhinitis (AR), six ASA-sensitive patients with chronic rhinosinusitis (CRS), and seven normal, nonatopic control subjects (NC). Nasal secretions from all three groups showed significant NCA (mean 157.1 +/- 54.0, 62.2 +/- 20.7, and 39.4 +/- 11.4% of FMLP chemotactic activity for AR, CRS, and NC subjects, respectively). Nasal secretions from patients with AR expressed significantly higher NCA (P < 0.02) than did secretions from NA patients. NCA was unchanged by heating at 56 degrees C for 60 min and was not susceptible to degradation by trypsin. Nasal challenge with Dermatophagoides pteronyssinus antigen induced clinical symptoms and resulted in significant increases in total protein and albumin concentrations in nasal lavages in AR patients, but failed to change the mean NCA activity for up to 40 min after the challenge. These results indicate that nasal secretions from both atopic and nonatopic patients express NCA, but its relation to allergic inflammation remains to be established. PMID- 8238797 TI - Comparison of two in vitro assays, RAST and CAP, when applied to the diagnosis of anaphylactic reactions to honeybee or yellow jacket venoms. Correlation with history and skin tests. AB - We compared the results obtained with a new specific IgE assay (Pharmacia CAP system) to those of RAST and intradermal skin tests (ST) performed in 87 patients with a history of generalized reaction to honeybee or yellow jacket venom. When CAP and RAST were compared with positive ST performed with honeybee venom, CAP sensitivity was not significantly higher (98%) than that of RAST (95%). When yellow jacket venom was tested, CAP sensitivity (93%) was clearly superior to that of RAST (40%). When we compared the specificities of RAST and CAP to bee venom, RAST was positive in 21% of the 24 subjects with negative ST, and CAP in 42%. Among the 29 patients with negative ST to yellow jacket venom, RAST was positive in 17% and CAP in 28%. These results do not reflect a lower specificity of CAP, because CAP positivities could be inhibited in vitro, and because, in three patients with a history of anaphylactic reaction (one to honeybee, two to yellow jacket), CAP was the only positive test confirming the clinical observation. Among the 53 patients who were able to identify the offending insect (honeybee, 31; yellow jacket, 22), the cause of the anaphylactic reaction was usually confirmed by ST and CAP: honeybee venom 97% for both ST and CAP; yellow jacket venom 82% for ST, 86% for CAP. This was not the case for RAST, which confirmed honeybee venom hypersensitivity in 87% and yellow jacket venom hypersensitivity in only 41%. Thus, CAP is both more sensitive and more rapid than RAST, without losing specificity. PMID- 8238798 TI - Loratadine and terfenadine in perennial allergic rhinitis. Treatment of nonresponders to the one drug with the other drug. AB - The efficacy of loratadine and terfenadine in perennial allergic rhinitis was evaluated in a double-blind, selected cross-over study consisting of two phases. During the first phase, 76 patients with perennial allergic rhinitis, 8-67 years old, were included in the study. Of these, 41 patients received loratadine 10 mg daily, and 35 patients received terfenadine 60 mg twice daily, for 2 weeks. According to symptoms and side-effects, 32 patients were classified as responders to loratadine, and 28 patients as responders to terfenadine. All observed symptoms were significantly reduced in both treatment groups, but with no significant differences between the two groups. Side-effects were few and mild. In patients with normal IgE, loratadine was significantly superior to terfenadine in relieving nasal secretion, whereas terfenadine was significantly superior to loratadine in relieving nasal congestion. In patients with increased IgE, patients treated with loratadine showed significantly greater reduction in sneezing than patients treated with terfenadine. A positive correlation between total IgE and reduction in overall symptoms was found for patients treated with loratadine, whereas a negative correlation was found for patients treated with terfenadine. During the second study phase, the nonresponders received the other drug for 2 weeks. All seven nonresponders to terfenadine responded to loratadine after crossing over, whereas four of nine nonresponders to loratadine responded to terfenadine. Nonresponders to one drug may respond to the other drug. Thus, more than one antihistamine drug should be tried in perennial allergic rhinitis if the first fails. PMID- 8238799 TI - The influence of IL-3, IL-5, and GM-CSF on normal human eosinophil and neutrophil C3b-induced degranulation. AB - The priming effect of interleukin-3 (IL-3), interleukin-5 (IL-5), and granulocyte/macrophage-colony stimulating factor (GM-CSF) on eosinophil and neutrophil degranulation was studied. Granulocytes were obtained from normal donors, and degranulation was induced by incubation with serum-opsonized Sephadex particles. The released amounts of eosinophil cationic protein (ECP), eosinophil protein X (EPX), myeloperoxidase (MPO), and lactoferrin (LF) were measured by radioimmunoassay (RIA). The effect of IL-5 was dose- and time-dependent, with a maximal enhancement of ECP and EPX release of 71% (P < 0.03) and 66% (P < 0.03), respectively. Neutrophil degranulation, however, was unaffected. IL-3 was marginally effective, whereas GM-CSF seemed to act as a secretagogue for both eosinophil and neutrophil degranulation. We conclude that IL-5 selectively primes eosinophil degranulation, whereas IL-3 and GM-CSF seem to act as secretagogues for eosinophils and neutrophils. The results indicate that IL-5 may be involved in the priming of eosinophils as observed in patients with asthma and hypereosinophilic syndrome (HES). PMID- 8238800 TI - Effect of a bronchial provocation test with house-dust mite on blood eosinophilia, eosinophil cationic protein, soluble interleukin-2 receptor, and interleukin-6 in asthmatic children. AB - Eighteen children with perennial asthma and allergy to house-dust mite (HDM) underwent a bronchial challenge with HDM. Before and 24 h after the test, a venous blood sample was taken to determine levels of eosinophils, eosinophil cationic protein (ECP), soluble interleukin-2 receptor (IL-2R), and interleukin-6 (IL-6). A histamine challenge was performed before and 24 h after the HDM challenge. All subjects showed an immediate asthmatic reaction (IAR). A definite late asthmatic reaction (LAR) was observed in 15 children, a probable LAR in two, and no LAR in one. Because of persistent bronchial obstruction (FEV1 < 70%), eight children were unable to perform a histamine challenge 24 h after the allergen challenge. These were the children with the lowest prechallenge provocation dose (PD20) of histamine. In the other 10 children, the mean PD20 histamine decreased after the HDM challenge (mean PD20 before was 0.56 mg/ml; after challenge it was 0.14 mg/ml; P = 0.007). After the HDM challenge, an increase was detected in the mean values of blood eosinophils (mean before was 446/mm3; mean after was 733/mm3; P = 0.002), ECP (mean before was 26.3 micrograms/l; mean after was 34.3 micrograms/l; P < 0.040), and IL-2R (mean before was 116.35 U/ml; mean after was 128.52 U/ml; P < 0.040). On the other hand, IL-6 remained unchanged after the HDM challenge (mean before was 9.47 pg/l; mean after was 9.70 pg/l; P = 0.360).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238801 TI - A prospective safety-monitoring study of immunotherapy with biologically standardized extracts. AB - We evaluated the safety of immunotherapy in 419 patients who attended our allergy department for treatment. They were suffering from rhinitis and asthma caused by sensitization to grass pollen or Dermatophagoides pteronyssinus. Immunotherapy was given by biologically standardized aluminum hydroxide adsorbed extracts according to a conventional schedule. Local reactions were recorded in 10.5% of the patients and systemic reactions in 4.8%. Only 0.37% of the doses administered were associated with systemic side-effects. We found that 84% of the patients who showed systemic reactions were asthmatic subjects (P < 0.01), and most of them were sensitized to D. pteronyssinus (71%). Side-effects occurred more frequently during the dose-increase period (P < 0.05). After 9482 doses had been administered, no anaphylactic shock or life-threatening reactions were registered. We believe the risk associated with immunotherapy to be drastically reduced when treatment is carefully monitored by skilled personnel. In such conditions, as shown by our study, immunotherapy is safe. PMID- 8238802 TI - Characterization and partial purification of the Croatian national standard Dermatophagoides pteronyssinus allergen extract. AB - Lyophilized Dermatophagoides pteronyssinus (Der p) allergen extract (AE) and partially purified Der p extract (PAE) were prepared and characterized. Partial purification of AE was performed by gel filtration on Sephadex G-100 and Sephacryl S-300. Crossed immunoelectrophoresis (CIE) disclosed the same precipitating lines in AE and PAE preparations. The relative potencies of AE and PAE were determined and compared with the WHO International Standard for Der p by the RAST inhibition method. The potencies were 6.5 x 10(5) IU and 1.5 x 10(6) IU, respectively. Biologic standardization by quantitative skin testing was performed with AE (20 selected patients) and PAE (12 patients). Median Ch was calculated by linear regression analysis (log-log model). One ampoule of AE contained 65,300 BU and 1 ml (vial) of PAE contained 166,000 BU. Der p AE could serve as a croatian national standard for further production of Der p allergenic extracts. PMID- 8238803 TI - The effect of immunotherapy on T-cell subsets in peripheral blood and bronchoalveolar lavage fluid in pollen-allergic patients. AB - The effect of immunotherapy (IT) on T-cell subsets in peripheral blood and bronchoalveolar lavage fluid (BAL) was examined in 15 patients with rhinoconjunctivitis and asthma caused by sensitivity to birch pollen. They were treated with IT for 3 years. Seven patients were treated with highly standardized birch-pollen extract (Pharmacia, Sweden). Eight untreated patients served as controls. Histamine challenge, blood sampling, and BAL were performed before (January, February), and at the peak of, the birch-pollen season (May). The subpopulations of T cells in peripheral blood and BAL fluid were investigated by immunocytochemistry and flow cytometry. During the birch-pollen season, the percentage of CD3+ and CD4+ cells of blood mononuclear cells in the IT patients increased significantly (P < 0.03 and P < 0.02, respectively). The percentage of CD8+ cells remained unaltered. In control patients, no changes of T-cell subsets in the peripheral blood were observed. T-cell subsets in BAL did not change during the season in relation to preseasonal values for either IT-treated or non IT-treated patients. PMID- 8238804 TI - Severe isolated allergy to Ficus benjamina after bedroom exposure. AB - Allergies to Ficus benjamina (Fb, weeping fig) have been described first as an occupational disease in plant keepers and later in clearly atopic subjects with known sensitizations to other inhalant allergens. We report a case of a hitherto nonatopic man with allergic rhinoconjunctivitis, contact urticaria, and asthma caused by a Fb kept in his bedroom for 18 months. Rubbing and skin prick test with native fig leaves and sap, and RAST to Fb were clearly positive. No other sensitization could be detected. After eliminating his Fb, he became asymptomatic within a few months. This case shows that private close contact to Fb can also induce allergy even in hitherto nonatopic subjects. Therefore, Fb should not be kept in bedrooms. PMID- 8238805 TI - Clinical improvement after unusual avoidance measures in the home of an atopic dermatitis patient. AB - A 27-year-old female office clerk with widespread atopic dermatitis (AD) since infancy appeared to be highly sensitized and exposed to molds, storage mites, and chicken feathers and moderately sensitized to house-dust mites and grass and birch pollens. Hardly any textiles were present in her home; that is, only 28 m2, which is less than 25% of the Dutch national average. The causal relationship between eczema and molds plus storage mites in this case of AD was strengthened by the positive effect of an unusual, multidisciplinary home-sanitation program involving cleaning of mineral surfaces and ventilation improvement. This home sanitation program led to a gradual drop of total IgE and clinical symptom scores to 21% and 13%, respectively, of the original values. PMID- 8238806 TI - Skin testing. The safety of skin tests and the information obtained from using different methods and concentrations of allergen. PMID- 8238807 TI - Glucocorticosteroids and rhinitis. PMID- 8238808 TI - Optimization of skin testing. II. Evaluation of concentration and cutoff values, as compared with RAST and clinical history, in a multicenter study. AB - In this multicenter study we evaluated the results of a previous study (part I) in a relatively large Dutch patient population, using the two previously tested allergens (house-dust mite and grass pollen) and two other standardized allergens (tree pollen and cat dander). The obtained skin test results were expressed as a histamine ratio and compared with RAST and clinical history (CH). The sensitivity and specificity were calculated at different cutoff values of the skin tests. The optimum cutoff values of 0.7 intracutaneous tests (ICT) and 0.4 skin prick tests (SPT) resulted in a predictive value for the detection of allergic sensitization of 83% (RAST) and 77% (CH), and 91% (RAST) and 86% (CH), for the ICT and SPT, respectively. As the ICT and SPT were performed in different centers, the results of these methods cannot be compared. No systemic side-effects of the skin tests were recorded. These results generally correspond well with the predictions regarding safety and predictive value of part I of this study, in which a limited number of patients was studied. In conclusion, through the use of a limited number of standardized allergens in a small group of patients, it may be possible to predict a safe and efficacious concentration for routine skin testing and to extrapolate from these results to other standardized allergens. PMID- 8238809 TI - Effects of interleukin-4 and interferon-gamma on the secretion of IgG4 from human peripheral blood mononuclear cells. AB - Whereas IgE antibodies are linked with allergy, IgG4 antibodies may reflect the state of immunity and protection against a particular antigen. It has been shown that interleukin (IL)-4 is required for induction of IgE synthesis. In order to elucidate the role of IL-4 in the production of IgG4 and to compare IgG4 and IgE regulatory processes, we quantified these immunoglobulin isotypes after in vitro culture of peripheral blood mononuclear cells (PBMC) in the presence of IL-4. The production of IgG4 was increased by IL-4 under the same conditions which are optimal for IgE production but not among PBMC from all donors, depending on the magnitude of spontaneous IgG4 secretion: IL-4 was effective only when the spontaneous secretion of IgG4 was < 7% of the total IgG secretion; it had no effect when spontaneous IgG4 production was > 7% of total IgG. The IL-4-induced IgE response was consistently obtained when IgG4 was < 7% of total IgG but was markedly diminished or absent when IgG4 was > 7% of total IgG. If Staphylococcus aureus strain Cowan 1 (SAC) was present during the 48-h preincubation step, spontaneous IgG4 production was increased, but the stimulatory effect of this mitogen on immunoglobulin production, including IgG4, was markedly blocked by the addition of IL-4. In contrast, IL-4-induced IgE synthesis was strongly blocked by the presence of SAC. Finally, secretion of IgG4 (spontaneous and IL-4-induced) was suppressed among cells from most donors by interferon-gamma (IFN gamma).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238810 TI - Major allergens of date palm (Phoenix dactylifera L.) pollen. Identification of IgE-binding components by ELISA and immunoblot analysis. AB - The IgE-binding components of date palm (Phoenix dactylifera L.) pollen were determined by ELISA and Western blotting in atopic patients in order to identify its major allergens. From a pool of previously identified allergenic fractions and sera from 15 skin-test-positive, atopic subjects, four components of 12, 14.4, 57, and 65-67 kDa were found to bind IgE in 80-93% of sera. Two other components of molecular masses 28-30 and 37-40 kDa also bound 60-80% of atopic sera. The immunologic specificity of date-pollen allergen that induced antibody response in sera of atopic patients was confirmed with ELISA. Furthermore, most of the reactivity in pooled positive atopic serum and antiserum raised in rabbits was eliminated after the sera were absorbed with the allergen. IgG immunoblot analyses showed varying degrees of cross-reactivity with common local allergens, notably Bermuda grass, but were generally of low intensity. These results indicate that date pollen has six major allergens with the 12, 14.4, 57, and 65 67 kDa bands binding 80-93%, and the 28-30 and 37-40 kDa bands 60-80% of atopic sera. We propose that these major allergens be assigned the notations "Pho d I" to "Pho d VI" in the order listed. PMID- 8238811 TI - Comparison of IgE-binding antigens in horse dander and a mixture of horse hair and skin scrapings. AB - Extracts of horse dander (HD) and horse hair and skin scrapings (HHSS) have been compared with respect to their content of proteins and carbohydrates. The protein content of HD is more than double that of HHSS, while the carbohydrate content is of the same order. SDS-PAGE and IEF, both combined with immunoblotting, and CIE/CRIE showed the IgE-binding ability of the proteins/glycoproteins present in the two extracts. SDS-PAGE/immunoblotting showed the presence of mainly the same IgE-binding bands in the two extracts. Nine were detected in HD, and seven in HHSS. Four of these were glycoproteins. The most important allergen was a glycoprotein with molecular mass of 27 kDa showing IgE-binding ability to the 16 patient sera available. Another important allergen with molecular mass of 67 kDa is probably horse albumin. IEF/immunoblotting showed the presence of a glycoprotein with IgE-binding ability at pH 3.2 in both extracts. Tandem CRIE showed that both extracts contained the same four allergens as major allergens. In addition, as we observed when analyzing both extracts in CRIE against 16 different patient sera, each extract contained allergens not present in the other extract. We may conclude from this investigation that commercial allergen extracts to be used in diagnosis and immunotherapy should include both horse dander and horse hair and skin scrapings in the starting material for the preparation of extracts. If not, allergens which may be of importance for some patients will be missing. PMID- 8238812 TI - IgE antibodies against bovine serum albumin in a case of eosinophilic gastroenteritis. AB - Eosinophilic gastroenteritis is a disease characterized histologically by an eosinophilic infiltration of the gut. The cause of this disease remains unclear, although both food allergy and food intolerance have been implicated in its pathogenesis. We report the case of a 22-year-old man in whom gastrointestinal symptoms first appeared in childhood, with involvement of mucosa and muscularis layers of stomach and bowel. He presented high IgE blood levels, and his prick test was positive to bovine, pig, and lamb sera. Immunoblots from calf, pig, and lamb sera, incubated with the patient's serum and revealed by autoradiography, demonstrated the presence of a 65-kDa protein band that was recognized by IgE antibodies but not by IgG. This band corresponded to bovine serum albumin, while IgE did not show reactivity with human albumin. These data suggest a possible role for IgE-mediated hypersensitivity mechanisms in the pathogenesis of eosinophilic gastroenteritis. PMID- 8238813 TI - Plasma histamine and bronchial reactivity in allergic asthma. AB - Histamine is an important mediator of allergic inflammation and bronchial hyperresponsiveness (BHR), a hallmark of asthma. Studies on the relationship between plasma histamine and BHR in allergic asthmatic patients have yielded controversial results. We therefore measured plasma histamine and bronchial reactivity in 30 nonsmoker volunteers taking no medication. Eleven were normal subjects; 19 were stable, mildly allergic asthmatic patients. Venous blood was taken to measure blood cells and basal plasma histamine by radioimmunoassay. After blood sampling, all subjects underwent a measurement of PC20M (concentration of methacholine causing a 20% fall in FEV1). Mean plasma histamine levels were 0.21 +/- 0.1 ng/ml and 0.44 +/- 0.3 ng/ml in normal and asthmatic subjects, respectively (P < 0.05). We found a significant increase of blood eosinophils and basophils in asthmatic patients, and a positive correlation between plasma histamine and circulating basophils. PC20M was greater than 16 mg in normal volunteers, and mean PC20M was 2.1 +/- 2 mg/ml in asthmatic patients. PC20M did not correlate with plasma histamine levels, but it did so negatively with blood eosinophils. The increased plasma histamine concentration in mildly atopic asthmatic patients might be a consequence of the high basophil releasability of atopics and the higher basophil counts in allergic asthma. Plasma histamine is thus unlikely to be a determinant of BHR in asthma. PMID- 8238814 TI - Capsaicin at the workplace. PMID- 8238815 TI - Olive pollen allergy in Bari, Italy. PMID- 8238816 TI - [Bordetella pertussis: current aspects and future prospects in public health]. PMID- 8238817 TI - [Experimental bases for the intestinal transplant]. PMID- 8238818 TI - [The surgical aspects of the digestive carcinoid tumor]. PMID- 8238819 TI - [Erythropoietin: current aspects]. PMID- 8238820 TI - [The evolution of ideas in the surgery of breast cancer]. PMID- 8238821 TI - [The problems of basic genetics seen from the clinical picture of genodermatosis]. PMID- 8238822 TI - [Reflections on childhood thinness]. PMID- 8238823 TI - [The promotion of immunizations: a discussion of vaccinal schedules]. PMID- 8238824 TI - The peri-operative management of surgical insertion and removal of the intravenous oxygenator device (IVOX). A report of nine cases. AB - Intravenous oxygenation (IVOX) is a new technique for augmentation of gas exchange in patients who require near maximal conventional ventilatory support. Patients who require IVOX are, by definition, critically ill with a high expected mortality. At present, these high risk patients must be transferred to the operating theatre for the IVOX device to be inserted. This report describes the anaesthetic problems associated with nine patients in whom an IVOX device was inserted and removed in our institution. The mortality was six out of nine patients; all deaths occurred with the IVOX device in situ. Three patients died within 6 h of insertion. Four patients were female. The patients' ages ranged from 14 to 76 years. There were few immediate ventilation changes in the first 4 h after IVOX insertion. Inspired oxygenation concentration was reduced in only one patient. Positive end-expiratory pressure was not reduced. Peak inspiratory pressure decreased in four patients. Arterial oxygen tension increased in four patients (range 0.1-2.5 kPa) and decreased in five (range 0.1-3.4 kPa). Arterial carbon dioxide tension increased in one patient (0.3 kPa) and decreased in eight (range 0.1-2.7 kPa). Inotropic support with adrenaline, dobutamine and noradrenaline needed to be initiated or increased in eight patients. Eight patients required 2-4 units of blood to be transfused during IVOX insertion or in the following 2 h. One patient suffered an asystolic cardiac arrest during the operation, but was resuscitated successfully. Three patients survived to have the IVOX removed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238825 TI - Effects of thiopentone, etomidate and propofol on beat-to-beat cardiovascular signals in man. AB - Analysis of beat-to-beat fluctuations of heart rate and blood pressure is a promising new approach to the clinical diagnosis and management of alterations in cardiovascular regulation. We investigated the effects of three induction agents on beat-to-beat heart rate and blood pressure fluctuations in man. Beat-to-beat fluctuations were analysed by spectral analysis. Two spectral areas are of main interest. One area is centred on the respiratory frequency and shifts with changes in respiratory rate (High Frequency area). These high frequency fluctuations in heart rate are caused by respiratory-induced blood pressure fluctuations, mediated by the vagus nerve through the baroreflex mechanism. Variability in the Low Frequency area, which occurs between 0.06 and 0.12 Hz is considered to be related to haemodynamic fluctuations due to the sympathetic baroreflex control loop of vasomotor activity and heart rate. Results from our study indicate that thiopentone, etomidate and propofol show considerable differences in their effects on beat-to-beat variability of heart rate and blood pressure. These differences can be explained by their specific effects on the cardiovascular system. PMID- 8238826 TI - Epidural infusions for nulliparous women in labour. A randomised double-blind comparison of fentanyl/bupivacaine and sufentanil/bupivacaine. AB - Sixty nulliparous women received epidural infusions in labour of 0.0625% bupivacaine containing either 2.5 micrograms.ml-1 of fentanyl or 0.25 micrograms.ml-1 of sufentanil, each starting at 12 ml.h-1. The duration of each stage of labour did not differ significantly between the groups nor did the mode of delivery. The quality of analgesia in the first and second stages of labour and at delivery was similar in the two groups and there were no significant differences in the bupivacaine dose requirements. In the fentanyl group, 90% of women required one or no top-ups compared with 87% in the sufentanil group. Five women in the fentanyl group and four in the sufentanil group developed motor blockade, limited to movement of the hip only. Six women (20%) in each group reported pruritus. There were no significant differences in Apgar scores, umbilical cord blood pH levels or neurologic and adaptive capacity scores at 2 or 24 h. Satisfaction with first and second stage analgesia was high with no differences between the groups. There were no significant differences in the incidence of postnatal symptoms with 52% of women reporting perineal pain and 45% localised backache. PMID- 8238827 TI - Effects of three different types of management on the elimination kinetics of volatile anaesthetics. Implications for malignant hyperthermia treatment. AB - The effectiveness of three types of management on the elimination kinetics of volatile anaesthetics was studied prospectively in 45 patients randomised to one of three groups. Patients were anaesthetised using isoflurane. Inspiratory and expiratory isoflurane concentrations were measured. After reaching a steady-state isoflurane concentration, the vaporizer was turned off. In group 1, only the fresh gas flow was increased from 40 to 120 ml.kg-1 x min-1. Patients in group 2, in addition to the increase in the fresh gas flow, had a charcoal filter connected in the inspiratory limb of the circuit. Patients in group 3 had the fresh gas flow increased and the anaesthetic machine and breathing system changed. There was a statistically significant difference in the isoflurane washout from the anaesthetic machines between group 1 (90% elimination time 39 [10] s) and group 2 (90% elimination time 25 [5] s) (p < 0.01). However, there was no significant difference in the isoflurane washout from the patients in any of the groups. Thus the use of a charcoal filter or a change of the anaesthetic machine and breathing system proved to be of no clinical advantage. PMID- 8238828 TI - A comparison between mivacurium and suxamethonium in children. AB - The neuromuscular effects of mivacurium were compared with those of suxamethonium in 69 children (aged 2-12 years), during nitrous oxide, oxygen and halothane anaesthesia in a randomised open study. Neuromuscular block was monitored by measuring the acceleration of the thumb caused by contraction of the adductor pollicis muscle after supramaximal stimulation of the ulnar nerve at the wrist using an Accelograph. End-tidal carbon dioxide was maintained at about 4 kPa in both groups. The mean times (95% confidence intervals) for T1:T0 ratio to decrease to 75%, 50%, 25% and 5% of control values were 50 (42-59), 62 (52-74), 83 (68-100) and 93 (46-108) s respectively for mivacurium and 18 (15-22), 26 (22 30), 32 (28-37) and 43 (38-49) s respectively for suxamethonium. The times for T1:T0 ratio to recover to 25%, 50% and 70% of control values were 615 (542-698), 769 (687-859) and 901 (820-993) s respectively for mivacurium and 196 (179-214), 216 (201-234) and 242 (216-259) s respectively for suxamethonium. The range of maximum block was similar for both drugs. The average time to reach maximum block was 143 s for mivacurium and 56 s for suxamethonium. Intubating conditions were similar in the two groups. PMID- 8238829 TI - The use of rocuronium (ORG 9426) in patients with chronic renal failure. AB - The pharmacodynamics of an initial dose of 0.6 mg.kg-1 rocuronium followed by three maintenance doses of 0.15 mg.kg-1 were studied during nitrous oxide/oxygen/isoflurane anaesthesia in patients with normal renal function (n = 12) and chronic renal failure (n = 12). The mean (SD) duration (min) of block after the initial dose was 28.0 (5.5) and 25.6 (11.7) respectively. The mean (SD) duration (min) of the effect of the three maintenance doses was 15.3 (4.0) and 14.2 (7.0); 17.3 (3.2) and 17.4 (8.7); 18.1 (2.8) and 19.1 (10.1) for the normal and renal failure patients respectively. The induced and spontaneous recovery indices were 3.7 (0.7) and 17.1 (6.9) in the normal group compared with 3.9 (0.5) and 19.0 (12.5) in the renal failure group and these values did not differ between the two groups. In this small study rocuronium appears to be suitable for patients with chronic renal failure. There is no evidence of prolonged block even when the drug is given in repeated doses for maintenance. PMID- 8238830 TI - Comparison of intubation following propofol and alfentanil with intubation following thiopentone and suxamethonium. AB - We have compared a rapid sequence induction of anaesthesia using propofol 2 mg.kg 1 and alfentanil 50 micrograms.kg-1, with a standard technique using thiopentone 5 mg.kg-1 and suxamethonium 1 mg.kg-1. The incidence of complications, and the quality of conditions for intubation, were similar in both groups. The patients receiving propofol and alfentanil showed a significant decrease in blood pressure and heart rate following induction (p < 0.01), whereas the group receiving thiopentone and suxamethonium showed a significant increase in blood pressure and heart rate following induction (p < 0.01). The use of propofol and alfentanil provided a satisfactory alternative to thiopentone and suxamethonium for a rapid sequence induction of anaesthesia. PMID- 8238831 TI - Accuracy of a computer-based anaesthetic audit system. AB - The accuracy of an anaesthetic computer-based audit system was assessed. In a retrospective survey of operations performed 4 months previously only 50% of the patients' notes could be found, and 40% of these notes did not contain an anaesthetic chart. A prospective survey was also performed. The accuracy with which the computer output reflected the anaesthetic technique was found to be 52% in the retrospective survey and 33% in the prospective survey. The authors conclude that audit systems should be audited before their generated data are used for decision-making. PMID- 8238832 TI - Rhabdomyolysis following life-threatening acute asthma attack. AB - The authors report a life-threatening, acute asthmatic attack in a young man, which was followed by rhabdomyolysis and acute myoglobinuria. A discussion of the possible role of the pharmacological treatment of asthma in initiating muscle damage is included, based on a review of the current literature. PMID- 8238833 TI - Acceleration of ventricular tachycardia following propofol in a patient with heterotopic cardiac transplant. Cardioversion of ventricular tachycardia in the native heart. AB - A 52-year-old Caucasian male underwent heterotopic cardiac transplant and subsequently developed a ventricular tachycardia in his native heart. The arrhythmia was successfully treated by cardioversion, despite an increased rate associated with induction of anaesthesia with propofol. The method of synchronised cardioversion is described and a possible hypothesis for the acceleration of ventricular tachycardia following induction is discussed. PMID- 8238834 TI - CS gas--implications for the anaesthetist. AB - Although the use of CS gas is illegal in the UK, an occasional patient exposed to its effects may be seen. We report the problems experienced with the anaesthetic management of such a patient. PMID- 8238835 TI - Respiratory depression following combination of epidural buprenorphine and intramuscular ketorolac. AB - A 48-year-old man who had undergone thoracotomy for carcinoma of the middle third of his oesophagus developed severe postoperative respiratory depression following intramuscular ketorolac 30 mg 2 h after 150 micrograms epidural buprenorphine. Summation of analgesia by drugs used in combination can have deleterious respiratory effects. PMID- 8238836 TI - Epidural catheters. Breaking and extraction forces. AB - The forces required to break and to remove catheters from epidural spaces were investigated. Provided the patient's back is fully flexed and a slow steady pull is applied to the epidural catheter, the extraction forces required at both thoracic and lumbar levels are well below the minimum force required to break the same catheters. PMID- 8238837 TI - The performance of two intensive care humidifiers at high gas flows. AB - High continuous flow breathing systems are now available to provide fresh gas flows well in excess of 100 l.min-1 in continuous positive airway pressure systems used for respiratory support. The performance of two commonly used intensive care humidifiers, the Kendal Conchatherm and the Fisher and Paykel FP310 have been assessed at flows of 50, 75, 100, 125 and 150 l.min-1. Their performance when using two humidifiers connected in parallel and in series was also studied. At a fresh gas flow of 100 l.min-1 the single Conchatherm gave an absolute water vapour concentration of 15.6 g.m-3 and the single FP310 11.9 g.m 3. At all flows the best results were achieved using two Conchaterm humidifiers in series. It is concluded that with high continuous flow breathing systems the humidification achieved with conventional humidifiers may be inadequate and it may be necessary to combine two humidifiers to obtain clinically useful humidification. PMID- 8238838 TI - Anaesthesia: cheap at twice the price? Staff awareness, cost comparisons and recommendations for economic savings. AB - We questioned 50 anaesthetic staff about the costs of consumables used in routine anaesthetic practice. By means of a questionnaire staff were asked for estimates of the cost of 28 drugs, fluids and disposables. The responses were more accurate than in previous surveys; 47% of all the estimated costs were within 50% of the actual costs and 75% were within 100%. The costs of relatively expensive items such as blood, laryngeal mask airways, enflurane and isoflurane were consistently underestimated whereas cheaper items such as disposable syringes and electrocardiograph electrodes were consistently overestimated. Using 'state of the art' consumables, an hour-long anaesthetic in 1992 cost 14.02 pounds compared with 1.24 pounds in 1959. However, allowing for inflation, the cost today is actually equivalent to the cost in 1959. Since 1980 there has been no consistent pattern in drug prices, with some prices remaining the same, others falling and some increasing; all are, however, cheaper in real terms. It is possible to make substantial savings by using the more expensive drugs judiciously and by encouraging the use of low-flow, closed-circuit anaesthetic systems. PMID- 8238839 TI - A technique of awake fibreoptic intubation. Experience in patients with cervical spine disease. AB - We report our experience of 58 patients undergoing awake fibreoptic intubation for cervical spine surgery. Topical anaesthesia was administered using a 'spray as you go' technique. The tracheas of 57 patients were intubated successfully. The mean time from starting topical anaesthesia to intubation was 16.1 min (SEM 1.0). The procedure was associated with cardiovascular stability, a low incidence of severe cough and had a high patient acceptance. There was a low failure rate amongst trainees. The minimum mean arterial oxygen saturation was 92.9% (SEM 1.1). There was no relationship between the time taken to intubation and the episodes of desaturation or the grade of laryngeal visibility at direct laryngoscopy. This method of producing topical anaesthesia for awake fibreoptic intubation is recommended. PMID- 8238840 TI - Rhabdomyolysis and bilateral diaphragmatic palsies following abdominal tamponade. PMID- 8238841 TI - Anaphylactoid reaction following local anaesthesia for epidural block. PMID- 8238842 TI - Fixed dilated pupils associated with alcuronium infusions. PMID- 8238843 TI - Mapleson D coaxial breathing system integrity testing. PMID- 8238844 TI - Simultaneous use of two laryngoscopes. PMID- 8238845 TI - Isoflurane, nitrous oxide and oxygen analgesic mixtures. PMID- 8238846 TI - Ondansetron and the prevention of postoperative nausea and vomiting. PMID- 8238847 TI - Drager Narkomed 4: machine fault. PMID- 8238848 TI - Pre-emptive analgesia. PMID- 8238849 TI - Use of alfentanil in association with intracranial disease. PMID- 8238850 TI - Clinical significance of the growth of micro-organisms in propofol. PMID- 8238851 TI - Blood sampling from paediatric arterial cannulae. PMID- 8238852 TI - Paediatric epidural catheters. PMID- 8238853 TI - Epidural mexilitine. PMID- 8238854 TI - EMLA cream for postcircumcision analgesia. PMID- 8238855 TI - Plasma concentrations of lignocaine after rectal administration. PMID- 8238856 TI - Follow-up of reported awareness. PMID- 8238857 TI - Severe pain from a subcutaneous haematoma following epidural analgesia in labour. PMID- 8238858 TI - Anaesthesia for the porphyric patient. PMID- 8238859 TI - Faulty catheter mount. PMID- 8238860 TI - Minitracheostomies made easy. PMID- 8238861 TI - The problem of partial competition in the quantitative characterization of interactions by competitive binding assays. AB - Binding expressions are derived and analytical procedures developed for the quantitative characterization of inhibitor binding that is only partially competitive with the interaction between an acceptor and the ligand that is being monitored. Two such situations are considered: (i) that in which the partial competition reflects binding of inhibitor to fewer acceptor sites than available to ligand; and (ii) that in which the partial competition reflects binding of inhibitor to acceptor sites in addition to those occupiable by ligand. The potential efficacy of the suggested analyses is then explored by their application to simulated data that span the likely range of experimental behavior. Quantitative analysis of the displacement of [3H]nitrobenzylthioinosine from cultured leukemic cells by an adduct of 5'-S-(2-amino-ethyl)-N6-(4 nitrobenzyl)-5'-thioadenosine with fluorescein-5-isothiocyanate is used to establish that the cells possess 6% fewer sites (150,000 cf. 159,000 sites/cell) for the fluorescent adduct than for the tritiated ligand, and that the binding is 10-fold weaker (binding constant of 0.28 cf. 2.8 nM-1). Corresponding analysis of results obtained with bovine aorta endothelial cells indicates that a 3-fold weaker interaction (binding constant of 1.1 cf. 3.3 nM-1) occurs between the fluorescent adduct and 79% of the cell sites accessible to the tritiated ligand. The present analytical procedures extend the utility of competitive binding assays for the quantitative screening of potential inhibitors by removing the inherent limitation of existing analyses that all acceptor sites be accessible to both the competing solute and the indicator ligand. PMID- 8238862 TI - Analysis of cholesterol and phospholipid hydroperoxides by high-performance liquid chromatography with mercury drop electrochemical detection. AB - High-performance liquid chromatography (HPLC) with reductive mode electrochemical detection on a mercury drop has been employed for the separation and determination of lipid hydroperoxides. Under the conditions used, baseline separation is achieved for three cholesterol hydroperoxide (ChOOH) standards, not only from one another, but also from two different phosphatidylcholine hydroperoxide (PCOOH) standards. Applying this method to a test system, photodynamically treated murine leukemia cells, we have identified and quantified a major family of overlapping PCOOHs and three ChOOHs, two of which are characteristic singlet oxygen adducts. In a typical separation, the detection limit is < 0.5 pmol for ChOOHs and < 50 pmol for more slowly eluting PCOOHs. In this respect, mercury drop detection outperforms all previously described electrochemical detection methods for lipid hydroperoxides and compares favorably with other HPLC-based approaches. However, in terms of equipment cost, relative simplicity of operation, and fewer potential artifacts, this method has a clear advantage over all other existing high-sensitivity methods. PMID- 8238863 TI - Detection of quantitative polymerase chain reaction products by hybridization on magnetic support with 125I-radiolabeled probes: quantification of c-myc copy numbers. AB - We present a technique for determining c-myc copy numbers that can be used as a prognosis index for some cancers. The method is based on the use of both competitive polymerase chain reaction and hybridization of amplified products. Coamplification was performed directly on cells with a synthetic oligonucleotide used as internal standard. It recognized the same primer set as the target. Coamplified products were captured on streptavidin magnetic beads as solid support using a 5' biotinylated primer. DNA immobilized on this support was denatured with alkali. Each coamplified product (target and reference gene) was further hybridized to two distinct specific oligonucleotide probes. Gene amplification levels were determined using a standard curve obtained by serial dilutions of peripheral blood lymphocytes run along with the experimental samples. This approach provides a rapid (less than 2 days) and reproducible method for evaluating c-myc gene copy number and may be used to quantify any gene. Moreover, its format allows for automation. PMID- 8238864 TI - Oligosaccharide composition of heparin and low-molecular-weight heparins by capillary electrophoresis. AB - The application of capillary electrophoresis to total compositional analysis of heparin and low-molecular-weight heparin samples has been studied. Optimum resolution of 17 defined oligosaccharides was obtained with the buffer system composed of 10 mM sodium borate and 50 mM sodium dodecyl sulfate at pH 8.81 and at a constant voltage of 20 kV. The ratio of oligosaccharide charge to the number of saccharide residues correlated with the migration time. For oligosaccharides having the same charge to saccharide ratio, the larger of the oligosaccharides eluted earlier. A hexasaccharide having a 3-O-sulfated glucosamine residue at the reducing end and arising from heparin's antithrombin III binding site, migrated in an unusual fashion. The limit of oligosaccharide detection was from 600 fmol to 1 pmol. Quantitative analysis could conveniently be performed on 10 pmol of an oligosaccharide sample. Oligosaccharide composition using capillary electrophoresis was obtained by nearly complete depolymerization of heparins with a mixture of heparin lyase I, II, and III. The analysis resulted in 95% mass balance for both heparin and low-molecular-weight heparin. Capillary electropherograms of heparin and different low-molecular-weight heparins depolymerized with heparin lyase I alone showed a high level of structural heterogeneity in the products formed. The oligosaccharide maps thus obtained might find use in fingerprinting the heparin and low-molecular-weight samples. PMID- 8238865 TI - Assays of retinoid dehydrogenases by phase partition. AB - Two modifications of an extraction assay for retinoid dehydrogenases are described. The first method involves the transfer of tritium from carbon-15 of [15-3H]-retinol or [15-3H]retinaldehyde to NAD, whereas in the second method, tritium from [3H]NADPH is transferred to all-trans-retinaldehyde. Since both versions of the assay involve the interconversion of water-soluble and -insoluble tritium-labeled compounds, a simple phase partition is sufficient to separate labeled products from labeled reactants. The assays are shown to provide reliable estimations of the reaction progress for three retinoid dehydrogenases of the visual system when compared to HPLC analysis of retinoid products or gel filtration analysis of pyridine nucleotide. The assays will be useful in studying retinoid dehydrogenases from other tissues and in principle can be modified for other dehydrogenase reactions with water-insoluble substrates. PMID- 8238866 TI - Analysis of myocardial plasmalogen and diacyl phospholipids and their arachidonic acid content using high-performance liquid chromatography. AB - A high-performance liquid chromatographic method for the assay of diacyl and plasmalogen (alk-1-enyl) phospholipid content and the determination of their fatty acid content from tissue homogenates is described. Myocardial phospholipids are rich in plasmalogens and have a high content of unsaturated fatty acids, including arachidonic acid, esterified in the sn-2 position. Using a three-stage HPLC assay we have analyzed the phospholipid subclass content and the amount of arachidonic acid esterified to these fractions extracted from isolated perfused rat hearts. After HPLC separation of total myocardial phospholipids, the phosphatidylcholine and phosphatidylethanolamine peak fractions are treated with phospholipase C to remove polar head groups and ultraviolet-absorbing benzoate derivatives are made. Separation and quantification of diacyl and plasmalogen content of the total phospholipids with nanomolar sensitivity is then achieved using isocratic elution with a silicic acid HPLC column. The separated plasmalogen and diacyl glycerobenzoates are then subjected to alkaline hydrolysis to remove fatty acids from the sn-2 position. The 2-(2,3 napthalimino)ethyltrifluoromethanesulfonate esters of the free fatty acids are then prepared and analyzed with subnanomolar sensitivity using reverse-phase chromatography with gradient elution. As plasmalogen-specific phospholipase A2 is activated during myocardial ischemia and comprises the majority of total phospholipase A2 activity in the heart, this methodology allows for a sensitive and complete determination of the changes in the mass of these phospholipids and their arachidonic acid content. PMID- 8238867 TI - Synthesis of alpha, beta-deuterated 15N amino acids using a coupled glutamate dehydrogenase-branched-chain amino acid aminotransferase system. AB - Gram amounts of various 15N-enriched L-amino acids have been synthesized using a coupled enzymatic system. Catalytic amounts of 15N-labeled L-glutamate are generated using (15NH4)2SO4, alpha-ketoglutarate, and glutamate dehydrogenase. The labeled glutamate in turn serves as an amine donor to an appropriate alpha keto acid using the Escherichia coli branched-chain amino acid aminotransferase, the subcloning and overexpression of which is described. In order to drive the reaction to completion the redox cofactor NADPH is regenerated using a glucose dehydrogenase system. Since the amino-transferase catalyzes exchange of the alpha hydrogen, carrying out this reaction in 2H2O gives rise to 2-2H,2-15N-labeled amino acids. Deuteration can be readily extended to the beta position as well by preexchanging the alpha-keto acids in basic 2H2O. The isotopically labeled amino acids are recovered in yields of 70-80%. PMID- 8238868 TI - Antigen-antibody binding and mass transport by convection and diffusion to a surface: a two-dimensional computer model of binding and dissociation kinetics. AB - The kinetics of binding and dissociation between a soluble analyte and an immobilized ligand on or near a surface are described numerically by an iterative computer model. The model is applied to a microflow chamber which is used for surface plasmon resonance measurements. It calculates diffusion perpendicular to the surface, flow parallel to the surface, and the interaction between any number of soluble and immobilized species. If the reaction between analyte and ligand is fast, binding and dissociation are influenced by the transport of the analyte to or away from the surface. In this case the measurement yields apparent association and dissociation rate constants which are not identical with the reaction rate of analyte and ligand. The transition between mass transport controlled processes and reaction-controlled processes is described and attention is drawn to possible misinterpretations of experimental binding and dissociation curves. The measurement of rate constants higher than allowed by the conventional technique can be performed by elution of the analyte with a second analyte of low molecular weight. PMID- 8238869 TI - Enhanced gel mobility shift assay for DNA-binding factors. AB - Gel mobility shift assays are commonly used to study DNA-binding factors involved in the regulation of constitutive, tissue-specific, and inducible genes. We found that addition of 3-[(3-cholamidopropyl)dimethylammonio]-propanesulfonate (Chaps, a zwitterionic detergent) at relatively high concentration (2.5% or more) to DNA binding reactions for four different factors (AP-1, SP1, GATA-1, and interferon alpha-regulated factor ISGF3) assayed in cell extracts greatly enhanced the signal for DNA-protein complexes (up to about 20-fold). The amplified signal for DNA-protein complexes so obtained was (at least in part) due to increased binding efficiency, as revealed by greatly reduced amounts of the free probes in the gels. The binding specificity, however, was not compromised. The fact that DNA protein complex formation with four different factors was stimulated by Chaps suggests that the enhancing effect of Chaps may be more general and not limited to certain types of DNA-binding factors. The results provide the basis for a highly sensitive assay for DNA-binding factors, which may be useful in several types of studies on such factors. Among other detergents tested, Chapso (another zwitterionic detergent), NP-40, and octylglucopyranoside (nonionic detergents) were found also to enhance the complex formation as tested for AP-1 binding, whereas sodium cholate and deoxycholate showed strong inhibition. PMID- 8238870 TI - Determination of apolipoprotein(a) isoforms by sodium dodecyl sulfate polyacrylamide electrophoresis and immunoblotting. PMID- 8238871 TI - Reverse transcription and PCR amplification of rare mRNAs immobilized on oligo(dT) cellulose. PMID- 8238872 TI - Isolation and purification of trehalose 6-phosphate from Saccharomyces cerevisiae. PMID- 8238873 TI - A method for checking the volume of solutions microinjected into Xenopus oocytes. PMID- 8238874 TI - Plasmid DNA preparation by heat treatment of Escherichia coli lysates. PMID- 8238875 TI - Development and application of a method for the detection, elution, and characterization of rat submandibular proteinases separated on isoelectric focusing gels reveals male/female differences. PMID- 8238876 TI - Optimization of nonradioisotopic single strand conformation polymorphism analysis with a conventional minislab gel electrophoresis apparatus. AB - We have developed a means of nonradioisotopic single strand conformation polymorphism (nonRI-SSCP) analysis and applied it to the detection of a point mutation in the human tumor suppressor gene, p53. The method does not require any particular facilities or apparatus, such as a laboratory for radioactive materials, a large gel unit for sequencing, or a semiautomated electrophoresis system. This technique comprises amplification of DNA fragments by the PCR technique with specific oligonucleotide primers, denaturation, and electrophoresis on neutral polyacrylamide gels in a conventional minislab apparatus. The SSCP patterns on electrophoresis were detected with a commercially available silver stain method. We also evaluated various electrophoretic conditions for nonRI-SSCP analysis, such as the gel concentration and buffer components. A tris/glycine buffer system gave better resolution of SSCP bands. The SSCP patterns of different sized DNAs could be analyzed in a gradient polyacrylamide gel. Thus, nonRI-SSCP analysis with a conventional minislab gel electrophoresis apparatus can be satisfactorily substituted for a commonly used RI-SSCP technique. PMID- 8238877 TI - Analysis of femtomole concentrations of alpha-ketoisocaproic acid in brain tissue by precolumn fluorescence derivatization with 4,5-dimethoxy-1,2-diaminobenzene. AB - An extremely sensitive fluorimetric HPLC method used to determine in vivo cerebral alpha-ketoisocaproic acid (KIC) concentration with less than 2 g of tissue (e.g., a single rat brain, approximately 1.8 g) is reported. Removal of unwanted lipids and amino acids and isolation of alpha-ketoacids in an optimal derivatization buffer are achieved by C18 solid-phase extraction of the acid soluble fraction of brain tissue. Quantitation of KIC to the femtomole level is accomplished by reversed-phase HPLC using 4,5-dimethoxy-1,2-diaminobenzene precolumn fluorescence derivatization and on-line fluorescence detection. These techniques are applicable to femtomole quantitation of other alpha-ketoacids in various tissue and blood matrices. Combination of this fluorimetric method with simple nonchromatographic procedures to measure in vivo cerebral [1-14C]alpha ketoisocaproic acid radioactivities in tissue provides estimates of specific activities. PMID- 8238878 TI - A microassay for measuring cytochrome P450IA1 and P450IIB1 activities in intact human and rat hepatocytes cultured on 96-well plates. AB - This study deals with the development of a sensitive and simple microassay for measuring cytochrome P450IA1 and P450IIB1 activities in intact human and rat hepatocytes. The method is based on the metabolism by cells cultured in microwells of appropriate substrates at noncytotoxic concentrations (8 microM 7 ethoxyresorufin and 15 microM 7-pentoxyresorufin). After incubation of the probes with the cells, the dealkylated resorufin formed and released into culture medium was quantified. To ensure the hydrolysis of the resorufin conjugates eventually formed, culture supernatants were incubated in the microwells with beta glucuronidase and arylsulfatase. The fluorescence was then read using a microplate fluorescence reader. A high correlation between the monooxygenase activity measured by this procedure and that measured by conventional procedures in the microsomal fraction of the same cells was found. The major advantages of this method are: (1) the small number of cells required; (2) a drastic reduction in assay time; (3) that the assay is performed in intact cells; and (4) the possibility of performing repeated assays with the same cell monolayer over a period of several days since no injury to cells is detectable during the assay. This method proved to be very convenient for studying cytochrome P450 induction by xenobiotics in primary cultures of human hepatocytes. PMID- 8238879 TI - Selective removal of free dodecyl sulfate from 2-mercaptoethanol-SDS-solubilized proteins before KDS-protein precipitation. AB - Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the discontinuous system of Laemmli is used world-wide for analytical and preparative gel electrophoresis of polypeptides. A minor but disturbing problem is the difficulty of concentrating highly diluted solutions and determining their protein content after 2-mercaptoethanol-SDS solubilization. We describe a solution to both of these problems, detailing a two-step procedure which takes advantage of the low solubility of potassium dodecyl sulfate (KDS). Removal of excess of SDS and 2-mercaptoethanol, and concentration of proteins from even a nanomolar solution, is achieved by a two-step KDS precipitation. Free dodecyl sulfate is precipitated in step one, while KDS-proteins are pelleted in the second step, allowing the thiol agents to be discarded with the supernatant. The effects of changing [SDS] and [KC1], temperature, and pH were studied to optimize the separation of free SDS from proteins. After final precipitation, the hundred- or thousandfold concentrated proteins can be suspended in a small volume of any required medium. The procedure allows protein determination by the Lowry method, peptide mapping of 2-mercaptoethanol-SDS-solubilized polypeptides, and all other analyses which are otherwise hampered by excesses of SDS and/or thiol reagents. PMID- 8238880 TI - Recombinant technology as an alternative to chemical peptide synthesis: expression and characterization of HIV-1 Rev recombinant peptides. AB - Recombinant DNA technology has been widely used for the production of proteins in recent years. In this paper, we describe the expression and the purification of two specific peptides corresponding to parts of the human immunodeficiency virus Rev protein. The strategy of this method relies on the chemical synthesis of a pair of two complementary oligodeoxynucleotides corresponding to the coding region of the peptide of interest and the subsequent cloning into a prokaryotic expression vector. Transformation of Escherichia coli with these synthetic gene constructs yielded high production levels of recombinant protein in the bacteria. The recombinant protein was composed of two moieties, one corresponding to an "affinity handle" and the second corresponding to the peptide. Chemical cleavage of the fused protein followed by a combination of affinity chromatography and rp HPLC led to rapid and convenient peptide purification. Peptide fused to the affinity handle as well as cleaved peptide were fully characterized by N-terminal microsequencing and mass spectrometry. The data presented demonstrate that although the major recombinant products had the expected amino acid composition, we detected unexpected processing such as alternative cleavage within the signal peptide, modified cysteines, and deamidations. These results emphasize the importance of the complete characterization of recombinant products by efficient analytical tools such as N-terminal microsequencing and mass spectrometry. PMID- 8238881 TI - An amplified assay for thiols based on reactivation of papain. AB - A sensitive spectrophotometric assay has been developed for thiol (sulfhydryl) groups using an inactive disulfide derivative of papain (papain-S-SCH3). The thiol-disulfide interchange reaction of a thiol with papain-S-SCH3 results in the stoichiometric formation of active papain (papain-SH). The reactivated papain catalyzes the hydrolysis of a chromogenic substrate, resulting in an amplified spectrophotometric signal proportional to the initial amount of thiol. A variety of thiols, e.g., cysteine, glutathione, penicillamine, cysteine methyl ester, and cysteamine, yield similar linear plots for the activity of papain vs the initial amount of thiol. An unknown concentration of a thiol is measured using a standard plot for the activity of papain vs the amount of thiol, obtained for the same thiol or for a similar thiol. Thiol groups on proteins and thiol groups of high values of pKa (2-mercaptoethanol, 3-mercaptopropanoic acid) can also be assayed using papain-S-SCH3 in the presence of excess cystamine. The assay is about 100 fold more sensitive than that using Ellman's reagent [5,5'-dithiobis(2 nitrobenzoic acid)]. A 0.4 microM solution of cysteine produces an absorbance change of 0.55 at 410 nm after 30 min in the assay, compared to a predicted change in absorbance of 0.0054 using Ellman's assay. PMID- 8238882 TI - Spectrophotometric determination of mimosine and 3-hydroxy-4-(1H)-pyridone--the toxic principles of Leucaena leucocephala. AB - A sensitive and selective spectrophotometric method for the estimation of the toxic factors mimosine and 3-hydroxy-4-(1H)-pyridone (DHP) has been developed based on the intense yellow-colored azodye formed with p-nitroaniline which showed a sharp absorption maximum at 400 nm. The method was optimized based on relative sensitivity of the reaction with various aromatic primary amino compounds and under different conditions of pH. Interference from a variety of structurally related compounds and phenols was tested and found insensitive to this method. The molar extinction coefficient at 400 nm for the azodye formed with mimosine was 5.31 x 10(4) M-1 cm-1 and that for DHP was 1.699 x 10(4) M-1 cm 1. The applicability of the method was tested using different plant extracts and recovery was found to be at 100 +/- 0.3%. The method is suitable for accurate estimation of both mimosine and DHP after paper chromatographic separation of extracts of different biological samples. PMID- 8238883 TI - A quantitative determination of singlet oxygen with horseradish peroxidase. AB - A singlet oxygen determination method based on the formation of dimethyl-(2,5)-2 methoxy-5-hydroperoxydihydrofuran (DMFO2) followed by acid hydrolysis with 0.1 N H2SO4 is described. Hydrogen peroxide is formed thereby as a product of hydrolysis which is then determined by the horseradish peroxidase-catalyzed formation of the homovanillic acid dimer. Since DMFO2 is a crystalline compound with sufficient stability it can be used as a primary standard for the determination. The peroxidase-catalyzed hydrogen peroxide determination allows detection of singlet oxygen concentrations as low as 10(-7) M. PMID- 8238884 TI - Methods for liquid- and solid-state CP-MAS NMR spectroscopy of untreated tissue biopsies. AB - We describe a method for NMR analysis of rapidly cooled or frozen biopsies and report its use on rat liver. Since the metabolic state of the biopsies can be expected to reflect the in vivo state, the method may be used as an alternative to the traditional examination of a perchloric acid extract of the biopsy. Perfusion-cooling of rat liver was applied as an efficient means of preserving the metabolic state. This method ensures very rapid cooling without interruption of the oxygen supply to the liver, and the results show that biopsies taken subsequently maintain energy metabolites near in vivo concentrations for at least 40-60 min. High resolution, natural abundance 13C liquid-state NMR spectroscopy could be carried out within this time frame on the untreated biopsy. In addition, the biopsy was frozen for 13C cross polarization-magic angle spinning solid-state spectroscopic examination, which was carried out at -40, -100, and -150 degrees C. The solid-state spectra allowed analysis of the relative glycogen content of the intact liver tissue, which showed good correlation with chemically measured glycogen on the same samples. Furthermore it was observed that the C1-carbon of glycogen in all liver samples splits into two resonances (5.4 ppm apart) in the solid state but not in the liquid state. This suggests that two conformational states of glycogen are populated, with rapid equilibration in the liquid state but no equilibration in the frozen state. PMID- 8238885 TI - Biotinylation of DNA on membrane supports: a procedure for preparation and easy control of labeling of nonradioactive single-stranded nucleic acid probes. AB - We have used M13 single-stranded DNA bound by uv to small pieces of nylon membrane for the synthesis of biotinylated single-stranded DNA probes. The labeling method requires a large fragment of DNA polymerase I and random hexanucleotides. There is no need for previous linearization of the template. The clean probe is removed from the membrane by a single wash step. The synthesized probe is completely free of unincorporated precursors. This makes possible the easy control of the reaction of incorporation of biotinylated analogues into the probe by simple staining on the filter, thus allowing evaluation of the efficiency of labeling. The DNA membrane can be stored for reuse. With the procedure described it is possible to biotinylate many DNA fragments in parallel, simultaneously controlling the efficiency of labeling in a time- and cost-saving manner. PMID- 8238886 TI - A method for defining the stages of low-density lipoprotein oxidation by the separation of cholesterol- and cholesteryl ester-oxidation products using HPLC. AB - A new high-performance liquid chromatographic system for the identification of some of the lipid oxidation products of low-density lipoprotein (LDL) oxidized by copper is described. Using a reversed-phase C-18 column and an isocratic solvent system of acetonitrile/isopropanol/water (44/54/2, v/v/v), a number of oxidized lipid moieties were resolved and detected simply by their 234-nm absorbance. The nature of several of these compounds was determined by chromatographic criteria, chemiluminescence, and mass spectrometry. The production of compounds within 4 h oxidation corresponded to the production of lipid hydroperoxides, the quantitatively most important of which is cholesteryl linoleate hydroperoxide, and to the rapid decrease in the cholesteryl ester content of LDL detected at 210 nm. More prolonged copper oxidation (up to 48 h) of LDL resulted in decreased quantities of lipid hydroperoxide moieties and increased amounts of a number of other, nonhydroperoxide, compounds. 7-Ketocholesterol and cholesterol linoleate hydroxide are two of the major products of prolonged oxidation. The detection of oxidation products correlates with the modification of LDL protein, permits a four-stage definition of metal-mediated LDL oxidation, and enables the calculation of a quantitative index of oxidation (lipoprotein oxidation index). This method will be generally applicable to cell- and copper-mediated oxidation, and will enable standardization of, and direct comparison between, different preparations of oxidized LDL. PMID- 8238887 TI - Measurement of cyclopentenyl cytosine 5'-triphosphate in vitro and in vivo by multidimensional high-performance liquid chromatography. AB - Cyclopentenyl cytosine 5'-triphosphate (CPEC-TP) is the active metabolite of the investigational drug cyclopentenyl cytosine (CPEC), a nucleoside analogue which exhibits noteworthy antineoplastic activity against several murine and human tumors in tissue culture, and which is now undergoing Phase I clinical trials. This study describes a method to measure the intracellular CPEC-TP levels in peripheral blood mononuclear cells (PBM cells) from patients treated with CPEC, without using radiolabeled drug. The method utilizes on-line multidimensional high-performance liquid chromatography (HPLC) with two columns of different retention mechanisms connected via an automated programmable switching valve. The elution fraction containing CPEC-TP is initially separated from cellular components using a gel sizing column (TSK-G2000-SW) and then rechromatographed by means of a reversed-phase column operated in an ion-pairing mode (YMC-A-312-ODS). The limit of quantitation of CPEC-TP by this method is 2.5 pmol per injection. When CPEC-TP levels were measured in PBM cells from 12 cancer patients after 20 h continuous infusion of CPEC at doses ranging from 3.5 to 5.9 mg/m2/h, the levels attained ranged from 1.4 to 13.4 microM (3.6 to 33.5 pmol/10(7) cells). However, wide variability in the concentrations of CPEC-TP achieved were evident at each dose and did not appear to correlate either with the CPEC dose or with CPEC plasma levels. This method was validated by a comparison of the quantitation of CPEC-TP in cultured PBM cells and Molt-4 cells (a human T-cell line adapted for growth in tissue culture) after incubation with both unlabeled and radiolabeled CPEC. PMID- 8238888 TI - Receptor-targeted transfection using stable maleimido-transferrin/thio-poly-L lysine conjugates. AB - Cell surface ligand-receptor systems provide favorable routes for DNA transfection due to target cell specificity, transfer efficiency, and low toxicity. Using the transferrin receptor system as a model, an approach to transfection is developed here within which involves the complexing of DNA to stable maleimido-transferrin/thio-poly-L-lysine conjugates. These studies establish the importance of precise stoichiometry for activity of ligand:poly-L lysine conjugates, as well as a chemistry for their controlled conjugation. Also considered quantitatively are effects of the following related parameters on the efficiency of receptor-mediated transfection: lysine polymer length, conjugate concentration, DNA:conjugate ratio, and treatment of target cells with chloroquine and desferrioxamine. Compared directly to standard procedures (electroporation, modified DEAE-dextran, lipofection, and modified Ca2PO4 protocols), transfection via this transferrin receptor-mediated system was > or = 10-fold more efficient, and essentially nontoxic to erythroleukemic F-MEL and J2E cells. Following transfection these cells retained the physiological capacity to undergo induced differentiation in response to dimethyl sulfoxide (F-MEL cells), or to erythropoietin (J2E cells), the natural hormonal regulator of erythropoiesis. Thus, an optimized approach to transferrin receptor-mediated transfection is developed which should be widely applicable for alternate cells and ligand-receptor systems both in vitro and in vivo. PMID- 8238889 TI - In vivo footprinting and genomic sequencing by ligation-mediated PCR. PMID- 8238890 TI - DNA adduct research with capillary electrophoresis. AB - DNA's central importance in all biological systems dictates a wide variety of DNA related research. For much of this research, the utilization of capillary electrophoresis (CE) can be of significant advantage. Open-tube CE yields excellent separations of DNA components, which can be monitored in a nearly "real time" manner by uv absorbance detection. Similar CE techniques enable the monitoring and optimization of enzymatic reactions. During DNA hydrolysis, both the reactants and the products can be simultaneously observed. DNA adducts (covalent additions of compounds to the DNA components) can similarly be analyzed by CE with uv detection. Many adduct nucleotides elute in a region of the electropherogram free from interferences due to the nonadducted components. Improved limits of detection are achieved when sample stacking techniques are utilized. Improved separations can result when the CE analysis involves micellar electrokinetic capillary chromatography. PMID- 8238891 TI - Elimination of protein band "edge tailing" in Weber-Osborn-type slab gel electrophoresis by glycerol. AB - In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) according to the method of Weber and Osborn, protein bands are often distorted by tailing at both ends, and the phenomenon is often called "edge tailing." This was eliminated by adding glycerol at a concentration of 10-15% (v/v) to the sample well gel supplemented to form the sample wells. This simple modification made the protein bands as sharp and straight as those in SDS-PAGE according to the Laemmli procedure. The linearity of the semilogarithmic plot of mobilities versus molecular mass was better than that obtained by Laemmli-type SDS-PAGE. PMID- 8238892 TI - Quantitation of electrostatic and hydrophobic membrane interactions by equilibrium dialysis and reverse-phase HPLC. AB - Equilibrium dialysis and reverse-phase HPLC have been used for the sensitive and precise quantitation of both electrostatic and hydrophobic interactions of peptides and small molecules with lipid bilayers. We show that hydrophobic solutes are rapidly and quantitatively released from lipid dispersions when loaded onto a C4 reverse-phase HPLC column equilibrated in water+0.1% trifluoroacetic acid and that the lipid molecules have no interfering effect on the chromatography. Peptides interacting electrostatically with bilayers are released quantitatively when a higher ionic strength buffer (water+2% ammonium acetate) is used. As little as 50 ng of solute can be accurately quantitated even in the presence of milligram amounts of lipid. We demonstrate the application of these methods to the hydrophobic interactions between indoles and lipid bilayers and to the electrostatic interaction between defensins, which are cationic antibiotic peptides, and anionic bilayers. The high sensitivity allows nondestructive quantitation of submicrogram amounts of precious solutes and the high precision allows the heat capacity change, an important thermodynamic parameter, to be obtained from the partitioning data. PMID- 8238893 TI - Analysis of trimethylsilylperoxy derivatives of thermally labile hydroperoxides by gas chromatography-mass spectrometry. AB - Qualitative and quantitative determinations of hydroperoxides are of central importance in many areas of biochemical research, but analysis of intact hydroperoxides by capillary gas chromatography-mass spectrometry (GC-MS) generally has not been possible due to the thermal instability of these compounds. It is demonstrated here that structurally diverse hydroperoxides can be converted to trimethylsilyl (TMS) peroxides and analyzed by GC-MS without significant thermal decomposition. The hydroperoxides investigated in this report include cumyl hydroperoxide, 2-phenylethylhydroperoxide, 13-hydroperoxy-octadeca 9,11-dienoic acid, 2,6-di-tert-butyl-4-hydroperoxy-4-methylcyclohexadienone (BHTOOH), and 2,4,6-trimeth-yl-4-hydroperoxycyclohexadienone (TMPOOH). A representative peracid, 3-chloroperoxybenzoic acid, was successfully analyzed as its TMS peroxyester. Chromatographic and mass spectral characteristics of the TMS peroxides and their corresponding TMS ethers were obtained and compared with thermospray HPLC-MS data for underivatized hydroperoxides. The utility of the GC MS assay for quantitating low levels of hydroperoxides in biological matrices was demonstrated by measuring BHTOOH and TMPOOH formed during the NADPH-dependent oxidation of alkylated phenols by rat liver microsomes. Overall, the results demonstrate that the advantages of GC-MS analysis (i.e., high chromatographic resolution, quantitative precision, high sensitivity, and the acquisition of structural information) can be applied successfully to intact hydroperoxides following trimethylsilylation. PMID- 8238894 TI - Multiple bands on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels of proteins due to intermolecular disulfide cross-linking. AB - Gel electrophoresis has been used extensively as an analytical technique to check the purity and to determine the molecular weight of proteins. Improved levels of detection of proteins by silver staining of the gels have made the technique more sensitive in detecting heterogeneity. We report here some interesting observations about the anomalous behavior of some proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This report is concerned with the appearance of multiple bands, after silver staining, on the SDS-PAGE gels of proteins which are shown by HPLC to be homogeneous. We also show the reasons for the appearance of bands that originate from 2-mercaptoethanol itself. PMID- 8238895 TI - Fluorescence properties of o-aminobenzoyl-labeled proteins. AB - Isatoic anhydride reacts with nucleophile groups of proteins to yield o aminobenzoyl protein conjugates. The fluorescence emitted by the chromophore decays in a multiexponential manner with average fluorescence lifetimes ranging from 9.9 to 10.7 ns. The steady emission anisotropy, measured upon excitation at 330 nm, is influenced by the molecular mass of the protein to which o aminobenzoyl is attached. The fluorescence properties of o-aminobenzoyl are suitable for rotational correlation time measurements of proteins smaller than 65 kDa. The technique of emission anisotropy can be used to detect interactions between proteins in solution, provided one of the proteins is labeled with o aminobenzoyl. PMID- 8238896 TI - A fast and accurate enzyme-linked immunosorbent assay for the determination of the DNA cleavage activity of restriction endonucleases. AB - We have developed an assay procedure to monitor the cleavage of DNA substrates by restriction endonucleases. This procedure uses DNA substrates that are labeled with biotin on one 5' end and with an antigenic group, e.g., fluorescein or digoxigenin, on the other 5' end. After incubation with the restriction enzyme, the reaction is stopped with EDTA and an aliquot is pipetted into the well of an avidin-coated microtiter plate. This immobilizes the unreacted substrate and the biotinylated cleavage product, whereas the other cleavage product labeled with the antigenic group is subsequently washed off. The unreacted substrate is detected by an enzyme-linked immunosorbent assay with an appropriate enzyme linked antibody. To test our assay we have measured the steady-state rate constants for cleavage of DNA by EcoRI yielding a kcat of 8.6 min-1 and a Km of 150 nM, which are close to values measured with other assays. The advantage of this assay is that it is not only fast and accurate, but also very sensitive. It allows for many samples to be analyzed in parallel and lends itself to automation. Furthermore, this assay can be designed as a competitive assay, when two substrates carrying different antigenic groups are used. The usefulness of such competitive assay is demonstrated by determining the influence of sequence context on the rate of DNA cleavage by EcoRI. PMID- 8238897 TI - Microassay of decarboxylation reactions in cultured cells. AB - The currently described methods for determination of decarboxylation reaction rates in cultured cells require large quantities of cells and often involve cell manipulation prior to assay. We describe a simple microassay for the rapid measurement of various decarboxylation reaction rates in intact cultured cells. The assay is based on the traditional measurement of 14CO2 generated from 14C labeled substrates. Key to the method is a novel modification of the standard petri dish. Pyruvate dehydrogenase, branched chain alpha-ketoacid dehydrogenase, alpha-ketoglutarate dehydrogenase, and ornithine decarboxylase activities were determined in adult cardiomyocyte cultures containing only 0.1-0.5 mg of protein per culture dish. Efficiency of 14CO2 collection ranged between 94 and 100%. Pharmacological enhancement or inhibition of pyruvate dehydrogenase activity was easily detected in the culture system. This new method simplifies the measurement of various decarboxylation reaction rates in cultured cells and allows rapid, reproducible measurements to be made on small numbers of cells without perturbation of the culture conditions or the cells themselves. PMID- 8238898 TI - Identification of a protein-tyrosine phosphatase from human platelet membranes by an immobilon-based solid phase assay. AB - The present investigations show that a 53-kDa platelet-membrane protein is a protein tyrosine phosphatase. Identification involved a novel methodology in which membrane proteins are resolved by polyacrylamide gel electrophoresis and then transblotted to a polyvinylidene difluoride membrane surface-labeled with [32P-Tyr](Glu4.Tyr1)n. Phosphatase activity appears as clear areas in the autoradiograph of the renatured 32P Western blot. Studies using this new solid phase system indicate that a 53-kDa platelet-membrane protein dephosphorylates [32P-Tyr](Glu4.Tyr1)n. Dephosphorylation is both time- and dose-dependent. The protein-tyrosine phosphatase antagonists orthovanadate and molybdate block dephosphorylation in a concentration-dependent manner. Inhibitors of protein serine/threonine phosphatases and of acid and alkaline phosphatases do not significantly affect enzymatic activity. The enzyme is active toward phosphotyrosyl proteins but not phosphoseryl proteins. Results from the solid phase phosphatase assays correlate with data from standard liquid-phase studies. A Lineweaver-Burk plot gives an apparent Km of 4.3 microM and an apparent Vmax = 168 nmol P(i) transferred/min/mg for the membrane enzyme towards the 32P-labeled polymer in a liquid system. Platelet lysate was shown by the solid-phase assay to possess a protein-tyrosine phosphatase of approximately equal to 50 kDa. The tyrosine phosphatase activities associated with a placental preparation, pure recombinant protein-tyrosine phosphatase 1B, and YOPR, the product of the YOP51 gene of Yersinia enterocolitica, which contains the C235R mutation, were also visualized by this technique. However, the catalytic activity of domain 1 of the transmembrane leukocyte antigen-related protein-tyrosine phosphatase was not detected using the solid-phase assay. PMID- 8238899 TI - Analysis of hydrophobic proteins and peptides by electrospray ionization mass spectrometry. AB - During the last decade mass spectrometry has become an essential tool for the analysis of peptides and proteins. Electrospray ionization mass spectrometry (ESIMS) is one of several recently developed techniques for the determination of accurate molecular masses of proteins, peptides, and other biopolymers up to > 100 kDa. Up to the present, analyses have been performed mainly on biopolymers that are soluble in aqueous solutions. Mass spectrometric analyses of very hydrophobic species, such as membrane proteins, have seldom been reported in the literature. This is mainly due to the incompatibility between most mass spectrometric techniques and detergents and/or salts which are required to retain such proteins in solution. Hydrophobic proteins (for example, bacterioopsin) and peptides are in general not soluble in the solutions (methanol/water or acetonitrile/water) typically used for ESIMS, and most detergents and chaotropes interfere with the analysis. We have developed sample handling protocols and solvent systems that are compatible with instrumental requirements and also are capable of retaining very hydrophobic peptides and proteins in solution. Chloroform/methanol/water mixtures were found to work well with, e.g., bacterioopsin, and also to be compatible with samples dissolved in hexafluoroisopropanol and 70-95% formic acid. PMID- 8238900 TI - Homogeneous model immunoassay of thyroxine by phase-modulation fluorescence spectroscopy. AB - We describe a homogeneous competitive model immunoassay for determination of thyroxine by multifrequency phase-modulation fluorescence. Using a nonradiative energy transfer transduction mechanism, B-phycoerythrin conjugated to thyroxine is the energy donor and a carboxymethylindocyanine dye conjugated to anti thyroxine antibody is the energy acceptor. Energy transfer from B-phycoerythrin to the acceptor results in a decreased lifetime and/or phase angle. The fluorescence lifetime change reflects the extent of energy transfer. In the competitive immunoassay format, the donor-thyroxine conjugate and an analytical sample of thyroxine compete for acceptor-antibody binding sites, resulting in a phase angle change which is dependent on the amount of thyroxine in the sample. Dose-response curves of phase angle versus thyroxine concentration are comparable to steady-state intensity curves. Since phase-modulation lifetime measurements are largely independent of total signal intensity, sources of optical interference are minimized. The potential for whole blood measurements exists since the energy transfer lifetime method can be extended to longer wavelengths. PMID- 8238901 TI - Derivative spectroscopy as a nonperturbative tool to detect the state of liposome entrapped sulbactam. AB - Sulbactam, a beta-lactam antibiotic, absorbs uv light at 273 nm when in alkaline media, whereas at neutral or acidic pH this peak disappears. Sulbactam-loaded liposomes, prepared by reverse-phase evaporation, were spectroscopically analyzed by the derivative mode measuring the peak-through amplitude between +258 and -285 nm, so that the spectral interference of the sample is eliminated. Taking advantage of this experimental approach, we could study the influence of different parameters on the dissociation state of the drug when entrapped in dipalmitoyl phosphatidylcholine liposomes. In particular, following the time course of sulbactam peak disappearance, we found that: (i) the effectiveness of protonating the sulbactam of the chosen buffers is acetate/acetic acid > succinate/succinic acid > citrate/citric acid; (ii) the rate of signal disappearance is influenced by the externally imposed pH and can be somewhat related to the dissociation state of the organic acids; (iii) as expected, the whole phenomenon is temperature dependent. The observations reported here might be the basis for quantitative permeation studies in synthetic and/or natural membranes using this methodology. PMID- 8238902 TI - Measurement of gene expression by multiplex competitive polymerase chain reaction. AB - We have developed a polymerase chain reaction (PCR)-based method to measure glutathione peroxidase (GSH-Px) mRNA levels. Expression was measured by multiplex competitive PCR amplification of (a) cDNA from GSH-Px and the "housekeeping" gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and (b) two internal standards consisting of single-base mutants of GSH-Px and GAPDH cDNA that cause either a loss (GSH-Px) or a gain (GAPDH) of an EcoRI restriction endonuclease recognition site. RNA extracted from a human papillomavirus-immortalized human bronchial epithelial cell line (BEP2D) was reverse transcribed. Serial dilutions of cDNA were PCR amplified in the presence of GSH-Px and GAPDH primers and quantified amounts of mutated internal standards. The amplified DNA was restriction digested with EcoRI and electrophoresed on an agarose gel stained with ethidium bromide, separating native from mutated products. Densitometry was performed to quantitate the bands. Our studies demonstrate that this technique measures the relative expression of GSH-Px to GAPDH precisely and reproducibly for studies done with the same master mixture and dilution of internal standards. Ratios of relative gene expression varied less than 25% from the mean. This technique will be useful to measure changes in gene expression, particularly when the amount of study sample is limited or the level of gene expression is low. PMID- 8238903 TI - Styryl-based wavelength-ratiometric probes: a new class of fluorescent calcium probes with long wavelength emission and a large Stokes' shift. AB - We describe the chemical synthesis and fluorescence spectral characterization of a styryl-benzothiazole probe which contains the Ca2+ chelating group BAPTA as an integral part of the chromophoric system. The visible absorption spectrum of this probe displays a dramatic shift in the long wavelength maxima from 508 to 407 nm upon complexation with Ca2+, with a Ca2+ dissociation constant of 1.5 microM. The emission maximum centered at 615 nm is well shifted from the absorption. The emission spectrum displays a small blue shift upon binding Ca2+, allowing this probe to possibly be used as an emission wavelength-ratiometric probe using a single-excitation wavelength. This probe is likely to be the first of a series of long-wavelength ratiometric Ca2+ probes whose structure can be modified for improved quantum yield or altered Ca2+ affinity. PMID- 8238904 TI - Improved method for hydrolyzing proteins and peptides without inducing racemization and for determining their true D-amino acid content. AB - A new method of hydrolyzing proteins and peptides without racemizing the amino acids has been developed. This method consists of performing a brief partial chemical hydrolysis for 15 min in 6 M HCl at 80-90 degrees C, followed by an enzymatic hydrolysis with pronase for 12-16 h at 50 degrees C, and finally an enzymatic hydrolysis with leucine aminopeptidase and peptidyl-D-amino acid hydrolase for 24 h. Using this new method the time required for complete hydrolysis of proteins is less than 3 days. The total hydrolysis averages 97 100%, and the amount of racemization of the amino acids is less than 0.002%. This method may then be used as a tool to easily determine the intrinsic D-amino acid content of peptides or proteins from animal or vegetable tissues. PMID- 8238905 TI - Self-association of Ca(2+)-binding peptides induced by lanthanide ions: a fluorescence study. AB - In this study a Ca(2+)-binding 14mer peptide was synthesized with the sequence GDKNADGCIEFEEL, allowing covalent attachment of sulfhydryl-reactive fluorescent molecules at position 7 of the 12-residue, metal-binding loop (underlined). This provided the opportunity to select donor molecules with suitable spectral characteristics for sensitized excitation of chelated terbium (Tb3+) or europium (Eu3+) ions. N-(1-Pyrene)-iodoacetamide and 7-diethylamino-3-((4' iodoacetylamino)phenyl)-4-methylcoumarin were attached to the peptide and titrations carried out with terbium or europium stock solutions. It was possible to observe lanthanide ion binding to the loop in stoichiometric quantities, but maximal lanthanide luminescence was achieved with a large excess of lanthanide present, due to metal-induced peptide association. Obtaining maximal lanthanide luminescence is important in the development of systems for use in sensitive clinical diagnostic and time-resolved luminescence-based immunoassay applications. PMID- 8238906 TI - Continuous and discontinuous assays for phosphotyrosyl protein phosphatase activity using phosphotyrosyl peptide substrates. AB - Assays for phosphotyrosyl protein phosphatase activity have been developed using the synthetic peptide Glu-Glu-Tyr(P)-Ala-Ala and its N alpha fluorenylmethoxycarbonyl derivative (Fmoc-Glu-Glu-Tyr(P)-Ala-Ala) as substrates. Conditions for monitoring the phosphatase-catalyzed hydrolysis of the former peptide, either spectrophotometrically or fluorometrically, are reported. These continuous assays are similar in sensitivity to published assays using phosphotyrosine as substrate. A discontinuous HPLC-based assay using the intensely fluorescent Fmoc-peptide as substrate is also described. This assay is comparable in sensitivity to assays using 32P-labeled substrates and is suitable for assaying low levels of phosphotyrosyl protein phosphatase activity in crude tissue extracts. PMID- 8238907 TI - An immune complex selective affinity column utilizing site-specific attachment of bovine conglutinin. AB - A simple method to isolate immune complexes by column chromatography is described. The immune complex affinity column was constructed by the site specific attachment of bovine conglutinin to agarose. Covalent attachment of conglutinin to agarose was achieved via hydrazone chemistry, which reacts aldehydes of oxidized oligosaccharides in the collagenous domain of conglutinin with hydrazide functional groups in the solid support. The constructed conglutinin affinity column captured complement-fixed model immune complexes of heat-aggregated human IgG and more classical complexes of chicken ovalbumin-anti chicken ovalbumin. Neither the nonfixed immune complexes nor their individual components were retained by the column. The captured material was specifically eluted with EDTA without the use of low pH. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot analysis, and protein sequence analysis of the eluates revealed the presence of the expected individual components, verifying that both antibody and antigen used to prepare the soluble immune complexes were recovered from the conglutinin column. The advantages of this approach over traditional methods of immune complex isolation and characterization are discussed. PMID- 8238908 TI - Protein-protein interaction studies using immobilized oligohistidine fusion proteins. AB - Using a human transcription factor, upstream stimulatory factor, as a model system, we developed a method for the rapid and efficient purification of proteins that interact with a cloned polypeptide expressed as a fusion with an oligohistidine domain. The complex between the oligohistidine fusion protein and its interacting partner was loaded onto a column of chelating resin charged with Ni2+. The bound complex could be eluted as a whole with high concentrations of imidazole. Alternatively, the secondary protein could be released separately using appropriate elution conditions. This procedure is both simple and efficient, and it presents distinct advantages over other affinity purification methods. In addition, this same method can be used to study protein-protein interactions. PMID- 8238909 TI - Detection of glycinamide ribonucleotide by HPLC with pulsed amperometry: application to the assay for glutamine: 5-phosphoribosyl-1-pyrophosphate amidotransferase (EC 2.4.2.14). AB - An assay method is described for measuring the catalytic activity of glutamine: 5 phosphoribosyl-1-pyrophosphate amidotransferase (EC 2.4.2.14), the first enzyme in the de novo purine biosynthetic pathway. Phosphoribosylamine, the unstable product of the enzymatic reaction, is trapped by glycinamide ribonucleotide synthetase (EC 6.3.4.13), the second enzyme in the pathway, to form the stable product glycinamide ribonucleotide. Glycinamide ribonucleotide is resolved by ion exchange HPLC at alkaline pH and quantified by the use of a pulsed amperometric detector. The assay is specific, reproducible, rapid (15 min chromatography, including column recycling and stabilization), convenient (no radioactive materials used), linear at a wide range of glycinamide ribonucleotide concentrations, and sensitive at subnanomole levels even with crude extracts. Both the glutamine- and ammonia-dependent activity of glutamine: 5-phosphoribosyl 1-pyrophosphate amidotransferase can be accurately quantified. PMID- 8238910 TI - Maleimide, iodoacetamide, indanedione, and chloromercuric spin label reagents with derivatized nitroxide rings as ESR reporter groups for protein conformation and dynamics. AB - The syntheses of eight nitroxide spin labels which bear maleimide, iodoacetamide, indanedione, or chloromercuric reactive groups and, in addition, a second substituent in the nitroxide ring are presented. The second substituent groups range from hydrophobic and hydrophilic esters to carboxylic acid and secondary and tertiary amine groups. The resulting spin labels are characterized with respect both to protein covalent modification and to the electron spin resonance spectral properties of the bound labels. The effect of the various substituents in the spin label on the reactivity toward the membrane-bound shark rectal gland and pig kidney Na,K-ATPase is described. The spectral differences between immobilized and mobile groups observed by electron spin resonance for the different protein-bound spin labels show that, by selecting an appropriate derivative for modification, a large range of different motional sensitivities of the reporter group can be obtained. Such different series of spin labels should therefore be useful for detecting mobility changes arising from conformational transitions in proteins by conventional electron spin resonance spectroscopy or for measurement of protein rotational diffusion using saturation transfer electron spin resonance spectroscopy. The chloromercuric series is found to be particularly useful because of the high reactivity, the lack of reversibility that potentially is associated with the Michael addition reaction, and the wide range of rotational mobility that is exhibited by the different derivatives. PMID- 8238911 TI - Measurement of plasma angiotensin II: purification by cation-exchange chromatography. AB - Measurement of plasma angiotensin II (AII) by radioimmunoassay (RIA) usually requires prior purification of the plasma to remove substances that cross-react in the RIA, most notably angiotensin III (AIII). Purification of AII is generally accomplished by solid-phase extraction (SPE) followed by reverse-phase HPLC, with tedious evaporation and resuspension steps in between, and requires collection of many HPLC fractions per sample for RIA. In this report, we describe a rapid two step SPE procedure for the purification of plasma AII, including an improved protease inhibitor cocktail for preventing the formation or degradation of AII in vitro. Plasma is first extracted on an S-Sepharose cation-exchange column, in which AII is separated from AIII by virtue of their difference in net charge, and then extracted on a C8 SPE column, without need for intermediate sample handling. The two-step SPE method is fast, results in only a single fraction for RIA per sample, and yields consistently high recoveries (77-86%) of AII, reducing the volume of plasma needed from 2 to 0.5 ml. Rat plasma was used in the present study, but the complete conservation of angiotensin peptide sequences (except angiotensinogen) in mammals suggests that this method will be applicable for other species including humans. In summary, the two-step SPE method offers the speed and simplicity of solid phase extraction while achieving a purity in AII (i.e., free of AIII) previously only obtained by laborious procedures involving HPLC. PMID- 8238912 TI - Analysis and purification of phosphorylated isoprenoids by reversed-phase HPLC. AB - Procedures were developed for analysis and purification of phosphorylated isoprenoids by reversed-phase HPLC. Dimethylallyl diphosphate (C5), geranyl diphosphate (C10), farnesyl diphosphate (C15), geranylgeranyl diphosphate (C20), and farnesylgeranyl diphosphate (C25) were cleanly separated on a C18 column by gradient elution with acetonitrile and 25 mM NH4HCO3. Reversed-phase HPLC was used to directly analyze extracts for phosphorylated isoprenoids and to follow the progress of enzyme-catalyzed reactions without having to hydrolyze the phosphate esters before analysis. Examples are provided for geranylgeranyl diphosphate synthase, geranylgeranylglyceryl phosphate synthase, and squalene synthase. The separations were run on preparative scales to purify products from the enzymatic reactions for mass spectrometry and to purify synthetic substrates. PMID- 8238913 TI - A simple method for determining kinetic constants of slow, tight-binding inhibition. AB - A general procedure is described for determining the kinetic constants of the slow, tight-binding inhibition of enzyme-catalyzed reactions by analyzing the data of initial and steady-state rate. All unknown parameters can be determined from several simple, sequential calculations. This method is simple and rigorous. It is also applicable to the special case of slow-binding inhibition, where the total concentration of inhibitor is much higher than that of the enzyme. PMID- 8238914 TI - Characterization of absorption and scattering properties of small-volume biological samples using time-resolved spectroscopy. AB - With time-resolved spectroscopy, we develop an experimental approach by sample substitution to measure the absorption (mu a) and reduced scattering (mu's) coefficients of small-volume biological samples. To investigate the method, small volume control samples are substituted into a large-size host medium during increases in the absorber (or scatterer) concentration of the host. By characterizing the deviation of the spectra taken with and without the sample, we determine the matching points where the sample and surrounding medium are optically identical. We show that this method can result in correct values of the mu a and mu's for the sample within 6% error if the matching conditions for both the mu a and mu's are fully realized. The results also indicate that this method can give approximate values of the mu a and mu's in a reasonable range if either the mu a or the mu's matching between the two media is realized. This method has been applied to the studies of absorption properties of a human finger and of scattering properties of yeast. PMID- 8238915 TI - A solid-phase assay for the binding of peptidic subunit association inhibitors to the herpes simplex virus ribonucleotide reductase large subunit. AB - The herpes simplex virus (HSV) ribonucleotide reductase is comprised of two nonidentical homodimeric subunits whose association is essential for enzymatic activity. In order to evaluate the affinity of a series of peptidic inhibitors with the enzyme's large subunit (R1), we have developed a sensitive solid-phase binding assay. The assay entails the use of a nonneutralizing antibody directed against the R1 subunit of the enzyme to immobilize either the native holoenzyme from HSV-1-infected cells or a recombinantly expressed HSV-2 R1 subunit. In either case, the radioiodinated peptidic inhibitor 125I-desamino-Tyr-(N-methyl) Val-Ile-Asp-(gamma-N,N-diethyl)-Asp-Leu demonstrated specific, saturable binding to the HSV R1 that could be competed by the nonapeptide Tyr-Ala-Gly-Ala-Val-Val Asn-Asp-Leu corresponding to the C-terminal sequence of the HSV ribonucleotide reductase small subunit (R2) or by recombinant HSV R2, but not by C-terminally truncated HSV R2 or murine R2. Our results provide direct evidence that inhibitors based on the carboxy-terminal amino acid sequence of HSV R2 compete with intact HSV R2 for a common binding site on HSV R1. The utility and sensitivity of this binding assay were further demonstrated by the ability to detect and discriminate ribonucleotide reductase inhibitors in the low nanomolar range. PMID- 8238916 TI - Distribution of ferrochelatase activity in density gradient separated bone marrow cells. AB - A simple, reproducible, and economical procedure for separating bone marrow cells into distinct populations and stages of maturation was developed and used to investigate the distribution of ferrochelatase activity in rat bone marrow cells. Density gradient ultracentrifugation of normal rat marrows on discontinuous arabinogalactan layers of 16.5-30.0% resulted in six distinct fractions enriched for myeloblasts, neutrophils, pronormoblasts, normoblasts, or lymphocytes. Segregation reflecting different stages of cell development was demonstrated. Neutrophils appeared at a greater density and were clearly separated from precursor myeloblasts. Normoblasts appeared at a greater density and were clearly separated from precursor pronormoblasts. Lymphocyte distribution within the gradient followed a normal curve (P < 0.05). A single gradient layer which consisted of greater than 80% lymphocytes was shown to be associated with significantly increased (P < 0.05) ferrochelatase specific activity (units/mg protein); the remaining cell populations did not differ significantly from each other in specific activity. PMID- 8238917 TI - Discontinuous electrophoresis of DNA: adjusting DNA mobility by trailing ion net mobility. AB - The use of a discontinuous buffer system, where a moving boundary separates ions of like charge but different ionic mobilities, for DNA electrophoresis may hold advantages over continuous zone electrophoresis in terms of resolution and electrophoresis time. Discontinuous buffer systems with calculated trailing ion net mobility were used to evaluate DNA mobility on gels of a constant pore size. Standard double-stranded DNA ladders and dideoxy sequencing ladders were electrophoresed on open-faced gels and standard sequencing gels, respectively. Trailing ion net mobility was systematically varied, while the leading ion mobility and concentration were kept constant. Decreasing trailing ion net mobility from 2.17 x 10(-4) to 0.59 x 10(-4) cm2 V-1s-1 generally led to increased DNA migration on both native and denaturing gels, allowing resolution of higher molecular weight DNAs with decreased electrophoresis time. However, on native open-faced gels, net trailing ion mobilities between 1.38 x 10(-4) and 1.76 x 10(-4) cm2 V-1s-1 had no differential effects for a 10-cm separation and kept DNAs smaller than approximately 75 bp stacked in the moving boundary and clearly resolved DNA between 100 and 600 bp. These results indicate that various DNA size ranges can be separated in short time periods by adjusting the net mobility of the trailing ion. PMID- 8238918 TI - Quantitative polymerase chain reaction analysis of mdr1 mRNA in multiple myeloma cell lines and clinical specimens. AB - We have designed a new polymerase chain reaction (PCR) protocol for the quantitation of mdr1 mRNA in cell lines and clinical specimens. This protocol uses an in vitro-generated RNA molecule as an internal standard. This synthetic RNA contains the same mdr1 primer sequences as the cellular mRNA, but yields a different-sized PCR product after amplification. Since a single primer set is used in quantitation, differences in primer efficiency are not a concern. We have used this assay to measure mdr1 expression in a multiple myeloma cell line, 8226/S, its drug resistant variants 8226/dox6 and 8226/dox40, and tumor samples from 10 patients with B-cell malignancies (9 multiple myeloma, 1 chronic lymphocytic leukemia). 8226/S does not express mdr1 mRNA. 8226/dox6 is 10-fold resistant to doxorubicin, and expresses 32 mdr1 mRNA/10 pg cellular RNA. 8226/dox40 is 140-fold resistant to doxorubicin, and expresses 890 mdr1 mRNA/10 pg cellular RNA. Seven of the 10 patients had levels of mdr1 mRNA expression below that seen in the multidrug-resistant, human multiple myeloma cell line, 8226/dox6. Three patients had levels of mdr1 expression comparable to those seen in 8226/dox6. No patient had levels of mdr1 expression close to that seen in 8226/dox40. Sample RNA integrity is assured by PCR analysis of a different, ubiquitous, cell cycle independent, histone variant, H3.3. This assay will be useful for studying low level mdr1 expression in cell lines and clinical specimens. PMID- 8238919 TI - Quantitative measurement of nonisotopically labeled polymerase chain reaction product. AB - A novel approach for evaluation of polymerase chain reaction product was described. A biotin-labeled DNA (or RNA) probe is hybridized to the digoxigenin labeled target PCR product. The hybrids are retained on an anti-biotin antibody coated plate. An anti-digoxigenin Fab conjugated with alkaline phosphatase is used for generating either a fluorescence or a chromagenic substance. Both fluorogenic and chromagenic systems can be used for quantitative measurement of the PCR product/probe hybrids. The method is specific, fast, nonisotopic, sensitive, quantitative, and practical. PMID- 8238920 TI - Quantitation of plasma endothelins. PMID- 8238921 TI - A method for the optical resolution of preparative scale quantities of DL-[N methyl-13C]carnitine. PMID- 8238922 TI - Adaptation of the periodate-resorcinol method for determination of sialic acids to a microassay using microtiter plate reader. PMID- 8238923 TI - Enhanced zymography of proteases. PMID- 8238924 TI - Preparation and characterization of covalently bound tetraphenylporphyrin-silica gel stationary phases for reversed-phase and anion-exchange chromatography. AB - A tetraphenylporphyrin-based stationary phase for high-performance liquid chromatography is prepared by attaching a p-carboxyphenyl derivative of the porphyrin to aminopropyl silica gel through an amide bond. This stationary-phase support is used for the reversed-phase separation of several polycyclic aromatic hydrocarbons and is shown to exhibit novel shape selectivity toward planar aromatic solutes. Subsequent metalation of the immobilized tetraphenylporphyrin with either tin(IV) or indium(III) results in a stationary phase with anion exchange characteristics. Both the tin(IV) and indium(III) metalloporphyrin-based stationary phases exhibit useful selectivity for the separation of aromatic sulfonates and aromatic carboxylates. PMID- 8238925 TI - Macroporous polymeric stationary-phase rod as continuous separation medium for reversed-phase chromatography. AB - A macroporous poly(styrene-co-divinylbenzene) rod has been prepared by a free radical polymerization of a mixture containing monomers, initiator, and porogenic solvent in the confines of a chromatographic column and used for the first time in the very fast reversed-phase HPLC of proteins. Characterization of the pore structure of the continuous rod by mercury intrusion porosimetry revealed a large volume of pores with a diameter of about 1 micron to pores below 100 nm. Size exclusion chromatography and scanning electron microscopy confirmed the unusual pore size distribution. The presence of large pores make the rod easily permeable to eluents, and therefore, the back pressure of the rod column is modest even at high flow rates. The efficiency of the polymerized column is almost independent of the flow rate. The slope of the line showing capacity factor vs composition of the mobile phase was determined for several proteins, and a gradient for the separation of their mixtures was developed. Excellent separation was achieved even at a high flow rate of 25 mL/min as documented by the resolution data. Tripling the length of the column did not improve the column resolution in protein separation. PMID- 8238926 TI - Application of computerized differentiation technique to remote-sensing Fourier transform infrared spectrometry for analysis of toxic vapors. AB - A differential method was developed for overcoming the difficulty in collection of background spectra when a remote-sensing FTIR is used during field measurements of air contaminants. A background spectrum was generated by introducing a delta lambda interval for every sampled data point of the spectrum. Temporal variations in spectral intensity caused by instability of the interferometer and changing IR beam configurations were greatly reduced through the use of the differential spectrum. Multicomponent analysis was performed by means of using a least-squares fit program with the differential spectrum. Three sets of spectra were evaluated by use of both the differential technique and the traditional method, which employs a clean background spectrum. There was no significant difference found between these two methods when a clean background spectrum could be used. However, in the case of field air monitoring system evaluations (as opposed to laboratory-based instrument evaluations), clean background spectra are frequently not available. Although the differential technique results in an increase in the limit of detection, results demonstrated that this method has sufficient sensitivity to detect and quantify both individual and mixtures of toxic compounds in the workplace. PMID- 8238927 TI - Sequence-selective biosensor for DNA based on electroactive hybridization indicators. AB - Deoxyribonucleic acid was covalently immobilized onto oxidized glassy carbon electrode surfaces that had been activated using 1-[3-(dimethylamino)-propyl]-3 ethylcarbodimide hydrochloride and N-hydroxysulfosuccinimide. This reaction is selective for immobilization through deoxyguanosine (dG) residues. Immobilized DNA was detected voltammetrically, using tris (2,2'-bipyridyl)cobalt(III) perchlorate and tris (1,10-phenanthroline)cobalt(III) perchlorate (Co(bpy)3(3+) and Co(phen)3(3+). These complexes are reversibly electroactive (1e-) and preconcentrate at the electrode surface through association with double-stranded DNA. Voltammetric peak currents obtained with a poly(dG)poly(dC)-modified electrode depend on [Co(bpy)3(3+)] and [Co(phen)3(3+)] in a nonlinear fashion and indicate saturation binding with immobilized DNA. Voltammetric peak currents for Co(phen)3(3+) reduction were used to estimate the (constant) local DNA concentration at the modified electrode surface; a binding site size of 5 base pairs and an association constant of 1.74 x 10(3) M(-1) yield 8.6 +/- 0.2 mM base pairs. Cyclic voltammetric peak separations indicate that heterogeneous electron transfer is slower at DNA-modified electrodes than at unmodified glassy carbon electrodes. A prototype sequence-selective DNA sensor was constructed by immobilizing a 20-mer oligo (deoxythymidylic acid) (oligo(dT)20), following its enzymatic elongation with dG residues, which yielded the species oligo(dT)20(dG)98. Cyclic voltammograms of 0.12 mM Co(bpy)3(3+) obtained before and after hybridization with poly-(dA) and oligo(dA)20 show increased cathodic peaks after hybridization. The single-stranded form is regenerated on the electrode surface by rinsing with hot deionized water. These results demonstrate the use of electroactive hybridization indicators in a reusable sequence selective biosensor for DNA. PMID- 8238928 TI - Examination of microdialysis sampling in a well-characterized hydrodynamic system. AB - The use of microdialysis sampling was examined in a well-characterized hydrodynamic system. A cross-flow microdialysis probe was designed in which the flow of both the dialysis perfusion solution and the sample solution could be carefully controlled. Dialysis membranes of cellulose (Cuprophan), cellulose acetate, and polyacrylonitrile (PAN) were examined in this system using hydroquinone as the test analyte. The permeability of the membranes to hydroquinone ranged from 1.72 x 10(-6) cm2/s for PAN to 2.97 x 10(-7) cm2/s for cellulose acetate. Determination of the dialysis fibers' recovery as a function of the sample flow velocity resulted in a rapid increase in recovery with increase in flow velocity. The recovery plateaued at high sample velocity. These results show that at low sample velocity diffusion through the sample solution is the rate-limiting step in recovery while at higher velocity transport through the membrane becomes rate limiting. Recovery for all three membrane types plateaued above sample velocities of 0.211 cm/s. This is well below the velocity of most biological fluids in which microdialysis sampling has been applied. This result supports previous reports that an in vitro calibration of microdialysis probes is appropriate for use in hydrodynamic environments in vivo such as the blood and bile. PMID- 8238929 TI - Determination of steroids in urine by micellar HPLC with detection by sensitized terbium fluorescence. AB - Efficient Forster energy transfer from steroid donors to a terbium ion acceptor occurs in aqueous solution when the two species are sequestered in sodium dodecyl sulfate micelles. The microorganized medium provides both proximity between donor and acceptor and protection from quenching of terbium fluorescence by water. Detection limits down to 100 pg have been achieved for steroids with an alpha,beta-unsaturated carbonyl group in the A-ring. This detection strategy has been used in HPLC separations, using either normal- or reversed-phase chromatography with postcolumn addition of detergent and/or terbium, or micellar chromatography with the detergent solution as the mobile phase. This last method proved to be superior, requiring no sample preparation and allowing direct injection of urine onto the HPLC column. PMID- 8238930 TI - Nonisotopic DNA detection system employing elastase and fluorogenic rhodamine substrate. AB - An alternative fluorescence-based method has been developed for the direct detection of small quantities of DNA in solution. In this system, a serine protease (elastase) is coupled to a DNA oligonucleotide through a disulfide linkage. A bis-(tetraalanine)-derivatized rhodamine molecule BZTAlaR) has been synthesized for use as a substrate. BZTAlaR is nonfluorescent in its derivatized form and shows negligible hydrolysis in solution. Cleavage of the tetraalanyl groups from the rhodamine portion of the molecule restores its fluorescence. Hybridization of the elastase-oligonucleotide conjugate to its target, capture of the conjugate-target complex with streptavidin-coated magnetic beads, addition of substrate, and subsequent detection of the target by fluorescence are accomplished in solution. Hybridization is rapid and specific, with over 90% of a target sequence successfully hybridized and captured. This method exhibits low background and an amplified fluorescent signal over time, resulting in a current detection limit of 0.49 fmol of elastase alone, or 2.64 fmol of conjugate, within 2 h. PMID- 8238931 TI - Structurally diagnostic ion-molecule reactions and collisionally activated dissociation of 1,4-benzodiazepines in a quadrupole ion trap mass spectrometer. AB - The ion-molecule reactions of various 1,4-benzodiazepines and dimethyl ether ions were studied with a quadrupole ion trap mass spectrometer. The methoxymethylene ions of dimethyl ether selectively react with 3-hydroxy-1,4-benzodiazepines (temazepam, oxazepam) to form (M+13)+ adducts by methylene substitution, and they react with 1,4-benzodiazepines that do not have hydroxyl substituents (diazepam, nordiazepam, nitrazepam) to form (M+15)+ adduct by a simple methyl cation transfer. These adducts are formed by elimination of methanol or formaldehyde, respectively, from (M+CH2OCH3)+ precursor ions. Ion-molecule reactions of model compounds with dimethyl ether ions suggest that the reactive site in the formation of (M+15)+ adducts is the imine functional group of the 1,4 benzodiazepines, while the reactive site for formation of (M+13)+ adducts involves a functional group interaction between the hydroxyl and carbonyl functional groups. Fragmentation induced by chemical ionization and collisionally activated dissociation provides further structural information for the differentiation of 1,4-benzodiazepines. Also, the gas-phase basicities of diazepam and temazepam have been estimated by bracketing techniques to be between 220.7 and 222.2 kcal/mol. PMID- 8238932 TI - Automatic precursor-ion switching in a four-sector tandem mass spectrometer and its application to acquisition of the MS/MS product ions derived from a partially (18)O-labeled peptide for their facile assignments. AB - We developed an acquisition processor unit to control MS1 and MS2 in a four sector tandem mass spectrometer simultaneously. The magnetic field strength of MS1 corresponding to the precise centers of the precursor ions, whose masses are determined by MS1 in advance, is recalibrated using an appropriate standard just before an MS/MS experiment, which allows consecutive acquisition of the MS/MS spectra from multiprecursor ions by B2/E2-linked scanning of MS2 synchronized with automatic B1 switching to the centers of each precursor ion on MS1. The accuracy of the automatic precursor-ion switching and the efficiency of consecutive B2/E2-linked scannings were demonstrated by measurement of a peptide mixture. The new MS/MS control system was further applied to rapid acquisition of the MS/MS spectra of isotopic ions of MH+ and (M + 3)+ with normal oxygen and (18)O at the alpha-carboxyl group of a partly (18)O-labeled peptide, which can be easily prepared by enzymatic hydrolysis of a protein in buffer containing 40 atom % H2(18)O. MS/MS analysis of an (18)O-labeled peptide by the present system allowed facile and reliable assignment of MS/MS product ions to a peptide sequence. The usefulness of the method was also demonstrated by measurement of some peptides that give MS/MS product ions incidentally observed at the same mass or assignable to some incorrect sequence. PMID- 8238933 TI - Synthesis of pentafluorobenzoic anhydride: a superior derivatizing agent for lipids. PMID- 8238934 TI - Trace elemental analysis at nanometer spatial resolution by parallel-detection electron energy loss spectroscopy. AB - Parallel-detection electron energy loss spectroscopy (EELS) combined with scanning transmission electron microscopy (STEM) and a field emission source provides an unprecedented sensitivity for elemental microanalysis. By deflecting the energy loss spectrum across a parallel detector and computing the difference spectrum from sequentially collected energy-shifted spectra, the effects due to detector pattern noise are nearly eliminated so that signals less than 0.1% of the background can be readily detected. Measurements on a series of glass standard reference materials show that EELS provides both high spatial resolution and trace sensitivity at the 10 atomic ppm level for a wide range of elements including the alkaline earths, 3-d transition metals, and the lanthanides. For analytical volumes with dimensions of the order of 10 nm, this translates into near-single atom detectability. PMID- 8238935 TI - Rapid biorecognition assay for herbicides in biological matrices. AB - A rapid assay capable of detecting several commonly used herbicides at nanogram per milliliter concentrations in biological fluids is described. The assay is based on inhibition of photosynthetic electron transport in spinach thylakoids by the target compounds with colorimetric detection using a redox dye. Using a microtiter plate format, high throughput assays of water, urine, and homogenized tissue were performed in minutes with minimal sample preparation. Detection limits of 3 ng/mL for atrazine, 3 ng/mL for diuron, and 1 ng/mL for terbutryn were observed. PMID- 8238936 TI - Sample cleanup by solid-phase extraction for the ultratrace determination of polychlorinated dibenzo-p-dioxins and dibenzofurans in biological samples. AB - A sample cleanup method for the isolation of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) by solid-phase extraction is described. A C18 bonded silica cartridge is used for the initial extraction and enrichment of the analytes. Subsequent cleanup is achieved with a dual-cartridge arrangement consisting of a bonded benzenesulfonic acid cartridge in series with a silica cartridge. A Florisil cartridge is employed for the final cleanup step. Sample preparation procedures used for blood plasma and animal tissues are described. The precision and accuracy of the method is validated by determining recoveries of 2,3,7,8-substituted PCDD and PCDF congeners spiked at various levels in blood plasma and in animal tissues. The effectiveness of the cleanup method is shown by the analysis of wild bird eggs which are highly contaminated with a variety of pesticides and polychlorinated biphenyls. The application of this cleanup method to human plasma containing PCDDs/PCDFs at parts-per-quadrillion levels and results of analyses by high-resolution mass spectrometry and low-resolution mass spectrometry are presented. The validity of this method is demonstrated by the comparable results obtained from this method and from other established methods. PMID- 8238937 TI - Evaluation of electrochemical parameters for an EC mechanism from a global analysis of current-potential-time data: application to reductive cleavage of methylcobalamin. AB - Simultaneous evaluation of electron-transfer rate constant, k degree, following chemical reaction rate constant, kf, electron-transfer coefficient, alpha, and standard potential, E degree', for electron transfer coupled to a following chemical reaction (EC mechanism) is described. A mathematical model for the current response to a potential step is developed by incorporating the appropriate concentration terms into the Butler-Volmer equation. Experimental current-potential-time (i-E-t) surfaces are fit to this model to evaluate the parameters. Fitting individual i-t or i-E curves did not yield unique parameter values whereas an i-E-t surface constituted by several i-t or i-E curves could be fitted to obtain unique values. A generalized kinetic zone diagram for the EC reaction is drawn by examining the limiting forms of the expression for current. Theoretical limits of measurable rate constants are estimated from the zone diagram. The three-dimensional electro-chemistry described above was used to study the reductive cleavage of methylcobalamin in dimethyl sulfoxide (DMSO) solvent and 0.1 M tetrabutyl-ammonium perchlorate supporting electrolyte. The parameters estimated are as follows: alpha = 0.552 +/- 0.004; k degree = 0.011 +/ 0.0015 cm s-1; kf = 1500 +/- 140 s-1; E degree' = -1.54 +/- 0.01 V. The rate constant for the following reaction, kf, in DMSO solvent is approximately 4000 fold faster than the similar process in aqueous medium. It is suggested that this enhancement is relevant to methyl group transfer in enzymatic reactions, e.g., methionine synthase, if the enzyme mechanism involves a reductive cleavage which produces a methyl radical. PMID- 8238938 TI - Initial studies of a new approach to the design and use of enzyme-based reactor/sensor systems: amperometric system for glucose. AB - This paper describes the development and evaluation of a new approach to the design and use of enzyme-based reactor/sensor systems (so-called "enzyme electrodes"). In the new approach, the reactor/sensor design is such that the measured response corresponds to reaction of all substrate in a fixed volume of solution. The result is that equilibrium-based measurements can be made, which in turn should result in advantages such as extended linear ranges and reduced dependencies on experimental variables such as enzyme activity, temperature, activators, inhibitors, etc. The concept was implemented with a glucose oxidase/electron-mediator reactor system immobilized on a glassy-carbon electrode operated in an amperometric mode. The reactor/sensor system was used in a thin layer (14 microns) cell such that the mean diffusion time of substrate (glucose) across the cell was very short (< 1 s) and the rate-limiting process was the chemical reaction at the reactor surface. In this way, it was possible to quantify the electrical charge corresponding to reaction of all the substrate in a fixed volume of solution perpendicular to the plane of the reactor system. Because the determined charge is dependent only on the total amount of substrate in the fixed volume, results exhibit linear range up to at least 2-fold the Michaelis constant and reduced dependency on pH relative to results obtained with steady-state responses from the same experimental system. A mathematical treatment is presented which yields equations that are consistent with time dependent responses for current and charge and which provide a rational basis for several data-processing options evaluated. PMID- 8238939 TI - Characterization of the chemical architecture of carbon-fiber microelectrodes. 2. Correlation of carboxylate distribution with electron-transfer properties. AB - Correlation of the chemical architecture of the surface of 10-microns-diameter carbon-fiber microelectrodes (illustrated by the fluorescence intensity of FITC labeled carboxylates) and the rate of electron transfer of the surface (illustrated by the intensity of the electrogenerated chemiluminescence of luminol) allows the development of quantitative relationships between the chemical structure of an electrode surface and its electron-transfer properties. A fluorescence microscope equipped with a Peltier-cooled charge-coupled device was used to image these electrode surfaces with submicron spatial resolution. The total fluorescence emission observed at electrochemically treated electrodes was higher than that of controls while the voltammetric behavior and integrated ECL intensity of luminol were very similar. Imaging spectroscopy with submicron spatial resolution was able to demonstrate the microscopic heterogeneity of these surfaces and to assess the effect of the production of carboxylates on the rate of electron transfer of luminol. PMID- 8238940 TI - Measurement of vanadium, nickel, and arsenic in seawater and urine reference materials by inductively coupled plasma mass spectrometry with cryogenic desolvation. AB - Addition of a small dose (2%) of H2 to the aerosol gas flow enhanced analyte signals by a factor of 2-3, which compensated for the loss of analyte signal that accompanied earlier efforts at cryogenic desolvation with inductively coupled plasma mass spectrometry (ICP-MS). Vanadium, nickel, and arsenic at microgram per liter levels in urine, river, and seawater reference materials were determined. The polyatomic ions ClO+, CaO+, and ArCl+, which normally cause severe overlap interferences for these elements, were attenuated to manageable levels by cryogenic desolvation. The samples were simply diluted with 1% HNO3 so that the chloride could be removed as HCl. The analytical results obtained for these standard reference materials agreed closely with the certified or recommended values. The detection limit ranges (3 sigma) obtained were 10-1000 ng L-1 for V, 0.03-20 micrograms L-1 for Ni, and 4-7000 ng L-1 for As in the original samples. The samples were introduced by flow injection to minimize clogging of the sampling orifice. PMID- 8238941 TI - Determination of double-bond position in some unsaturated terpenes and other branched compounds by alkylthiolation. AB - The electron-impact mass spectra of alpha, beta-bis(methylthio) derivatives of certain terpenes and other compounds with branched alkenyl groups contain diagnostic peaks that can be used for locating the position of the double bond in the parent compound. In contrast to the spectra of dimethyl disulfide (DMDS) derivatives obtained from compounds containing CH=CH type double bonds, which show two predominant fragment ions, most of the compounds examined in the current investigation showed only one predominant fragment ion, arising from that part of the molecule which possesses the more substituted carbon of the original double bond. For example, all the derivatives from compounds with an isopropylidene moiety showed a base peak at m/z89 which can be attributed to the formation of a [(CH3)2C=SCH3]+ fragment. The application of the DMDS procedure to naturally occurring terpenes is discussed. PMID- 8238942 TI - Analytical applications of catalytic properties of modified cyclodextrins. AB - This paper describes the evaluation of the catalytic properties of modified cyclodextrins for analytical applications. The beta-dimethylcyclodextrin was modified by adding one and two imidazolyl groups at carbon three positions. The modifications produced enhancements of catalytic activity for the hydrolysis of p nitrophenyl acetate at neutral pH by factors of 1000 or more relative to the unmodified cyclodextrins. The catalytic properties of the monosubstituted cyclodextrin were evaluated for the quantification of p-nitrophenyl acetate in the concentration range of 10-90 mumol/L. Results obtained by equilibrium, initial-rate, and error-compensating predictive kinetic methods were compared. The equilibrium and predictive kinetic options yielded virtually identical results, with linear changes with concentration throughout the range studied and severalfold larger than the initial-rate option and dependencies on temperature, pH, and catalyst concentration that are 5-10-fold smaller than the initial-rate option. PMID- 8238943 TI - Ultrahigh resolution matrix-assisted laser desorption/ionization of small proteins by Fourier transform mass spectrometry. AB - Recent research has demonstrated that matrix-assisted laser desorption/ionization (MALDI) is feasible for Fourier transform mass spectrometric analysis of biomolecules with masses in excess of 50,000 Da. Here, the effects of electrostatic deceleration times and laser energy upon mass resolution are reported. It is demonstrated that optimum deceleration times for singly-charged MALDI-generated protein ions ranging in mass from 2627 to 29,000 Da are a linear function of m1/2 when a 9.5-V decelerating potential is used. Furthermore, higher resolution is obtained with laser fluences close to the threshold for MALDI. Slow metastable decay of molecular ions in the absence of co-matrix is demonstrated for melittin and bovine insulin. It appears that the resolution enhancing effect of co-matrix may result from slowing molecular ion unimolecular decomposition rates sufficiently to allow infrared emission to compete with metastable decay, thus providing the requisite population of long-lived ions for high mass resolution. A spectrum of bovine insulin molecular ion with mass resolution of 30,000 is presented, together with several spectra of lower mass proteins with mass resolution in excess of 100,000. Detection of a doubly-charged carbonic anhydrase trimer ion with a mass of 87,000 Da is reported. PMID- 8238944 TI - Mathematical treatment of electrophoretically mediated microanalysis. AB - A new concept in reaction-based chemical analysis is introduced and theoretically described. By utilization of the variability in electrophoretic mobilities among charged species, spatially distinct zones of chemical reagents can be electrophoretically merged under the influence of an applied electric field. Electrophoretically mediated microanalysis (EMMA) exploits this phenomenon as a basis for chemical analysis utilizing capillary electrophoretic systems. EMMA is described in terms of the four stages required for reaction-based analysis: (1) analyte and analytical reagent metering; (2) initiation of reaction; (3) control of reaction conditions and product formation; (4) detection of species whose production or depletion is indicative of the concentration or quantity of the analyte of interest. The method is illustrated by the enzymatic oxidation of ethanol to acetaldehyde by alcohol dehydrogenase with the concurrent reduction of NAD+ to NADH monitored at 340 nm. Experimental results for both substrate and enzyme determinations are shown to agree with the presented theory. PMID- 8238945 TI - Engineering protein orientation at surfaces to control macromolecular recognition events. PMID- 8238946 TI - Purification and analysis of drug residues in urine samples by on-line immunoaffinity chromatography/high-performance liquid chromatography/continuous flow fast atom bombardment mass spectrometry. AB - An automatic system for the on-line extraction and analysis of diethylstilbestrol in the urine of rats and calves is described. Extraction was done by injecting samples directly into an immunoaffinity column containing antidiethylstilbestrol antibodies bound to a Sepharose matrix, and analysis was done by on-line high performance liquid chromatography with ultraviolet and continuous-flow fast atom bombardment mass spectrometry detectors. The system, consisting of one injector, two switching valves, and three pumps, was operated under computer control and allowed to perform a complete analysis of a sample in 28 min. An accurate quantitation by isotope dilution was also possible, by the use of deuterated diethylstilbestrol as internal standard. The sensitivity of the method, using selected-ion monitoring of the molecular ion of diethylstilbestrol, was 2 ng/mL, injecting 1 mL of urine sample. Results obtained from analyzing the urine of rats and calves treated with diethylstilbestrol are presented. PMID- 8238947 TI - A quest for oleandrin in decayed human tissue. AB - This forensic case taught us several lessons. First, there is a need for improved sample cleanup and treatment of severely decayed tissue samples when trace determinations of target analytes are needed. With the exception of a few reports the literature is lacking in information with regard to the most modern sample preparation techniques. Second, the coupling of LC/LC with tandem MS provides an effective means of "on-line" samples cleanup for complex sample matrices. The improvements in selectivity shown in Figure 3 reveal the analytical power available when these techniques are combined. Third, once we decided to use LC/LC/MS/MS, we were able to analyze more than 50 samples in a semi-automated fashion over approximately three days. The reliability and ruggedness of the combined techniques and equipment suggest this approach may have merit for common applications in which large numbers of biological samples (e.g., plasma and urine) must be analyzed. As a postscript, when this project was completed we proposed that the use of antibodies for isolating oleandrin and its relatives might be a more selective means for trace enrichment of the target analytes. For example, a high-pressure immunoaffinity column could have been coupled on line as column 1 in Figure 4. After pumping a relatively high volume of aqueous tissue extract through an immunoaffinity column during trapping and trace enrichment conditions, the column could be rinsed with phosphate-buffered saline. Then the pH could be lowered to unfold the antibody protein and allow release of the trapped analyte from this column with subsequent trapping on column 2 in Figure 4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238948 TI - Changes in the peritoneum during the development of the testis, epididymis and ductus deferens in the pig. AB - The development of the peritoneal folds of the testis, epididymis and ductus deferens of the pig prior to testicular descent was studied in 18 to 82 days-old embryos/fetuses. The parietal attachment of the mesonephros, mesonephric and paramesonephric ducts and gubernaculum constituted the urogenital mesentery. This could be divided in a cranial and a caudal part. The first fixed the mesonephros and had a mesogonad as a secondary fold. The second had two branches, one laterally to the umbilical artery belonging to the gubernaculum (Plica gubernacularis) and the medial to the artery contained the meso- and paramesonephric ducts. The mesogonad was related caudally with Plica gubernacularis. The fold of the meso- and paramesonephric ducts could also be divided in two parts. The caudal was a primary component of the urogenital mesentery, while the cranial was first ventral and later lateral to the mesogonad (mesorchium) and appeared as a secondary fold of the urogenital mesentery (cranial part). A vascular fold contributed to separate the suspensory ligament of the testis from the developing mesorchium. The proposed meaning of the terms used might be useful for comparative studies. PMID- 8238949 TI - The anatomy of the cloacal bursa (bursa of Fabricius) in the helmeted guinea fowl (Numida meleagris galeata). AB - The cloacal bursa (bursa of Fabricius) in the guinea fowls appeared either as an oval blind sac with a short thick stalk in one group or had a pointed cranial blind end with a slightly bulging middle part that was followed by a thick caudal stalk in the other group. Both groups of bursae originated from the proctodeal wall of the cloaca and were placed dorsal to the rectum. The average length of the bursa was 18 mm while the average width at the mid section was 15 mm. The internal surface showed about 12-14 primary folds. Histologically, the outline of the bursa was well established by day 18 of incubation. The primary folds had also been formed. Lymphocytes had already been encountered within the framework of the bursa at this day. The epithelium bordering the tunica propria was composed principally of two layers of cuboidal cells. Epithelial buds had also formed and some were already detached from the epithelial lining. The blood vessels present were positioned just beneath the outer covering. At day 19 of incubation, most of the epithelial buds had two layers of cells arranged in a circumscribed manner while a few had three layers of cells. Blood vessels had increased in number and were deeper placed inside the bursa than previously. At day 20, the cells of the upper layer of the epithelium were dorsoventrally flattened and stained paler than the cells of the lower layer. It was possible to distinguish the cortex from the medulla and the basement lining between both zones was distinct. Tiny vesicles within the cytoplasm of the epithelial cells at the mucosa and follicles were observed. Macrophages were also observed within the gland. At day 21, blood vessels were observed in the cortex of the follicles. The maximum number of primary folds (14) had been formed. At day 22, several follicles had severed connections with the mucosal epithelium. The mucosal lining had dropped to a single layer of cells in some areas. Goblet cells were observed amongst the mucosal cells. A plasma cell had first appeared. By day 25, dead cells had increased quite in number and there was also an increase in number of medium and small-sized lymphocytes within the gland. By day 26, the upper layer of the surface epithelium was composed primarily of tall columnar cells with numerous large vacuoles. Macrophages had suddenly increased within the thin interfollicular spaces and most of them were crowded internally with various sizes of debris.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8238950 TI - Retinal photoreceptor fine structure in the red-tailed hawk (Buteo jamaicensis). AB - The retinal photoreceptors of the red-tailed hawk (Buteo jamaicensis) consist of rods, single cones and double (unequal) cones present in a ratio of about 2:1:5. In the light-adapted state, the rods are slender elongated cells with outer segments that reach to the retinal epithelial (RPE) cells. The inner segment displays an ellipsoid of mitochondria, plentiful polysomes, some rough ER and Golgi zones. The rod nucleus is located deep within the outer nuclear layer and the synaptic spherule displays both invaginated (ribbon) and superficial (conventional) synaptic sites. Single cones show a thin tapering outer segment, a large electron lucent oil droplet at the apex of the inner segment and an ellipsoid of mitochondria. Double cones consist of a larger chief member which displays a thin tapering outer segment and an electron dense oil droplet as well as a smaller accessory cone which shows no oil droplet, an ellipsoid and a paraboloid of glycogen. As in the single cone, polysomes, RER and Golgi zones are also noted in the inner segments of both members of the double cone. Near the external limiting membrane the chief and accessory cones show membrane specializations indicative of junctions on their contiguous surfaces. All cone photoreceptors are of a smaller diameter than is normally reported for avian species. Both single and double cones display several invaginated synapses as well as numerous superficial synaptic sites. PMID- 8238951 TI - Intramuscular distribution of fibre types in the gluteus medius of the horse: a histochemical analysis. AB - The results from a recent study by Bruce and Schurg (1990) indicate that the gluteus medius (Gm) of the horse is a complex muscle subdivided by an internal tendinous sheet into two regions (dorsal and ventral) which may subserve different functions. In the present study, both regions were found to contain a similar proportion of histochemically-determined muscle fibre types. Fibre type composition did not generally vary over the length of the muscle at the same relative depth. In contrast, there was a significant tendency for the percentage of type I fibres to increase gradually in accordance with depth. These histochemical results do not suggest that the two regions of the Gm assume distinct functional roles; they do, however, indicate that at a given relative depth the muscle fibre type distribution is relatively uniform over the length of the muscle. Nevertheless, care should be exercised in sampling and interpreting data obtained from limited biopsy of equine Gm, due to the substantial histochemical variation of fibre type composition as a function of sampling depth. PMID- 8238952 TI - Regularities and irregularities in the structure of the seminiferous epithelium in the domestic fowl (Gallus domesticus). I. Suggestion of the presence of the seminiferous epithelial cycle. AB - A cellular association demarcated by two perpendiculars which were drawn between adjacent bundles of elongate spermatids from the tubular lumen to the basement membrane, was made the unit of histometrical observation in this study (provisionally called a "column"). Cell counting revealed that the average numbers per column of various types of germ cells do not show any significant differences among 5 fowls and between paired testes. The frequency of spermiogenic steps (numbered 1-8) was investigated in each column. A definite and common pattern was found in the frequency distribution in the 5 fowls observed. A relationship between spermiation and younger spermatid steps was also investigated in each column. The spermiation was found at different steps, but most frequently at step 2 (30.6%). Based on these observations and referring to other author's information, an average time interval between two successive spermiations was calculated roughly at 3.3 +/- 1.2 days. Theoretically, this value is equal to an average length of one epithelial cycle. Such a variable cycle may have caused irregular cellular associations in this species. PMID- 8238953 TI - Regularities and irregularities in the structure of the seminiferous epithelium in the domestic fowl (Gallus domesticus). II. Co-ordination between germ cell associations. AB - It was investigated in the domestic fowl, whether a mechanism for a synchronous start of the proliferation of the differentiating spermatogonia in a transversely cut seminiferous tubule (called the "co-ordination" between germ cell association) operates effectively in this species. A trapezoid-shaped cellular association demarcated by two perpendiculars from the tubular lumen to the basement membrane was the unit of observation (provisionally called a "column" Each column is composed of a bundle of elongate spermatids and the neighbouring, various types of germ cells). The extent of closeness of the co-ordination was examined in a total of 2,650 columns, based on the continuity of the spermiogenic step-number of the youngest generation of spermatids within adjacent columns. In no case did all columns in a tubular cross section show the same-step spermatids. In most cases (50.2% of 2,650 columns), the same number was kept in only 2 to 6 adjacent columns. The arrangement of different but consecutive numbers (e.g., 1-2 3 or 1-2-1) was observed in a total of 882 columns. The co-ordination mechanism, therefore, is considered to work, albeit incomplete, among a limited number of adjacent columns in this species. PMID- 8238954 TI - [Morphostructural study of the lingual papillae in the buffalo (Bubalus bubalus)]. AB - The morphostructural characteristics of lingual papillae in 30 adult buffaloes were studied by SEM. In this species, dorsal lingual surface papillae shows different shape so that their classification is complicated. Topography and morphology of conical and vallate papillae presents significant aspects. Particularly, the conical papillae have a different morphological organization when they are localized over the side of the tongue apex or over dorso-lateral tract of the tongue body. The functional aspects are discussed. PMID- 8238955 TI - Anatomical studies on the spinal cord segments of the impala (Aepyceros melampus). AB - The anatomy of the spinal cord segments was studied and recorded for the impala. The root attachment lengths were greatest at C3, T10 and L3 cord segment levels in the respective regions. As to the root emergence length the greatest lengths were observed at C7, T10, L5 and S1 cord segment levels respectively. The interroot interval was longest at C2, T8 and L1 segments respectively. The longest cord segments were C2, T13, L2 and S2 segments. The widest cord segments of their respective regions were C7, T1, L5 and S1 cord segments. As to segment volume C3, T13, L2 and S1 were the most voluminous cord segments in the respective cord regions. Statistical analysis revealed a high correlation among all of the study parameters suggesting a high degree of multicolinearity. Gross anatomical relationships concerning the location of the spinal cord segments with respect to the vertebrae were studied. The cord segments C1, T1-T4 and L1-L3 were within their vertebral limits. In the impala the spinal cord terminated at the midlevel of S4 vertebra. PMID- 8238956 TI - The arterial system of the wood mouse, Apodemus sylvaticus: the pelvic region. AB - In this paper we describe the origin, arrangement and variations of the major pelvic arterial trunks in the wood mouse, Apodemus sylvaticus. Seventy-six specimens have been analyzed by injection of coloured latex solution through the left ventricle of the heart and subsequent dissection of the pelvic region. Measurements of the origin of the major arterial trunks have been performed taking into account the number of anterior and posterior branches arising from its origin. Only the values of the internal iliac artery and the pudendoepigastric trunk show significant differences according to the number of previous branches. In general terms, the pattern of the pelvic arteries in the wood mouse agrees with those reported to for other rodent species. Nevertheless, some differences have been detected, especially in the origin of the major arterial trunks and the constitution of the internal iliac artery. PMID- 8238957 TI - Emergence of endothelial and hemopoietic cells in the avian embryo. AB - During organogenesis, endothelial cells develop through two different mechanisms: differentiation of intrinsic precursors in organ rudiments constituted of mesoderm associated with endoderm, and colonization by extrinsic precursors in organs constituted of mesoderm associated with ectoderm (Pardanaud et al. 1989). On the other hand, both types of rudiment are colonized by extrinsic hemopoietic stem cells. In the present work we extend our former study by investigating the hemangioblastic (i.e. hemopoietic and angioblastic) potentialities of primordial germ layers in the area pellucida during the morphogenetic period. By means of interspecific grafts between quail and chick embryos, we show that splanchnopleural mesoderm gives rise to abundant endothelial cells, and to numerous hemopoietic cells in a permissive microenvironment, while somatopleural mesoderm produces very few cells belonging to these lineages, or none. Thus we confirm that the angioblastic capacities of the mesoderm differ radically, depending on its association with ectoderm or endoderm. Furthermore, at this embryonic period, both endothelial and hemopoietic potentialities are displayed by splanchnopleural mesoderm. However the site of emergence of intraembryonic hemopoietic stem cells appears spatially restricted by comparison to more widespread angioblastic capacities. PMID- 8238958 TI - 3-D observation of N-cadherin expression during cardiac myofibrillogenesis of the chick embryo using a confocal laser scanning microscope. AB - It is not yet understood whether cell adhesion molecules play an active role in early cardiac morphogenesis or not. We present here the spatial and temporal expressions of N-cadherin and its relationships to actin filaments during looping (7- to 13-somite stages) of the chick embryonic heart tube observed by means of a confocal laser scanning microscope. Serial optical tomograms were obtained from the whole-mounted heart tubes stained with antibody to N-cadherin (fluorescein conjugated) and phalloidin (rhodamine-conjugated). Three patterns of N-cadherin expression were observed during looping; a belt-like pattern, speckled pattern, and clumped pattern, corresponding to adhesion belt, nonjunctional cell contact and early intercalated disks, respectively. At the 7-somite stage, myocytes expressed N-cadherin as adhesion belt and nonjunctional cell contact. At the 8- to 10-somite stages, the clumped pattern of N-cadherin was detected before striated myofibrils appeared. Myofibrils began to develop across the clumps to form transcellular networks in the outer layer, and to form circumferential alignments in the inner layer. These results suggest that N-cadherin is responsible for the connection of myofibrils between the neighboring myocytes, and the alignment of the two layers in the developing heart tube. PMID- 8238959 TI - Evidence for intussusceptive capillary growth in the chicken chorio-allantoic membrane (CAM). AB - The aim of our investigations was to test whether the chicken chorio-allantoic membrane (CAM) could be an adequate in vivo model for a new mode of capillary growth, originally described in the rat lung and termed intussusceptive microvascular growth. According to that concept the capillary system does not grow by sprouting of vessels, but expands by insertion of transcapillary tissue pillars or posts which form new intercapillary meshes. In the present study, we observed slender transcapillary tissue pillars with diameters around 1 microns in the CAM by in vivo microscopy, and analyzed their ultrastructure by transmission electron microscopic investigation of serial sections. The pillars corresponded in size to those previously described in rat lung microvasculature. On day 7, the pillar core contained endothelial-, endothelial-like cells and collagen fibers, and on day 12 additionally chorionic epithelial cells. As a hypothesis we propose that slender cytoplasmic extensions of endothelial cells, heavily interdigitated in the post area and often projecting into the vascular lumen, could initiate the first step of pillar formation, i.e., interconnect opposite capillary walls. During both stages of development endothelial-like cells were observed in close relationship with the pillars. These cells seem to be relevant for tissue post completion and growth, as they were found to invade the core of the pillars. From the localization of the interendothelial junctions in the post region, a certain similarity to the concept proposed for the lung can be found. The observations confirm that the CAM is a very suitable material for the in vivo investigation of intussusceptive capillary growth. PMID- 8238960 TI - Sensory nerve endings in the beak skin of Japanese quail. AB - This study is concerned with the distribution and ultrastructure of sensory nerve endings in the beak skin of adult Japanese quail (Coturnix coturnix japonica). The following nerve endings were found: free nerve endings, clusters of dermal Merkel nerve endings, Herbst corpuscles and Ruffini corpuscles. The latter were found only in the dermis of the tip of the upper beak. The remaining endings were present in the skin of all areas of upper and lower beak. Free nerve endings were supplied by either thin myelinated axons or unmyelinated C-fibers and were localized in the dermis close to the basal layer of the epidermis. Merkel cells formed clusters (up to 50) localized below and between the epidermal cones of the beak skin. Disc-shaped thickenings of nerve endings were squeezed between individual Merkel cells. Small Herbst corpuscles were found in the dermis close to the epidermal cones of the beak skin. Large Herbst corpuscles occurred in deep layers of the dermis. The Ruffini corpuscles were cylindrical in shape (80 microns x 400 microns) and arranged in groups of up to ten corpuscles. Each corpuscle was surrounded by an incomplete fibrous capsule. PMID- 8238961 TI - Developmental pathways of vertebral centra and neural arches in human embryos and fetuses. AB - The ossification pathways of both vertebral centra (i.e., vertebral bodies) and neural arches were studied in human embryos and fetuses (CR-length between 38 and 116 mm). A clearing and double-staining method for whole embryo or fetus, using alcian blue and alizarin red S, allowed an easy and precise detection of the morphology of the whole vertebral column and every single vertebra. Both cartilaginous and bony components were clearly visible. Different temporal and topographical patterns of ossification were shown for the centra and arches; the latter were respectively proximal-distal (i.e., bidirectional from a defined starting tract in T10-L1) and cranial-caudal (i.e., monodirectional). The patterns could be related to the morphogenetic processes of other structures (i.e., muscles and nerves). Moreover, the numerical survey of ossification centers provided a possible parameter for the determination of the fetal developmental age. This could be useful in the study of pathological conditions. PMID- 8238962 TI - Ultrastructure of the developing stomach in human embryos. AB - Ultrastructural development of the stomach was studied by light, scanning electron and transmission electron microscopy, using 19 human embryos at Carnegie stages from 14 to 23 (6.8-28.0 mm in crown-rump length, 5 to 8 weeks of gestation). The precise time of appearance of differentiated characteristic structures was examined electron microscopically. The first gastric pit, with radially arranged epithelial cells beneath which the basement membrane bulged into the mesenchyme, was observed on the lesser curvature at stage 22. Although the mesenchymal condensation which would develop into the inner circular muscle layer appeared at stage 18 onward, cytoplasmic myofibrils were not observed until stage 22. Nerve fibers were first observed at stage 16, and at later stages they gathered into bundles to form a nerve plexus external to the developing inner circular muscle layer. On the basis of accurate timing of the appearance and the mode of development of these structures, possible relations between developing gastric layers were discussed. Histocytochemically, glycogen or other carbohydrates were demonstrated in the cytoplasm of the gastric epithelium throughout the stages examined. These carbohydrates were localized mainly in vacuole-like spaces in the basal part of the epithelial cells. This subcellular localization, and the amount of carbohydrate, did not change significantly during the observed embryonic period. In the serosa, carbohydrates were not detected at stages 14 and 15, but observed consistently within the vacuoles in the cytoplasm from stage 17 onward. No other layer of the embryonic stomach had detectable carbohydrates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8238963 TI - The Splotch mutation interferes with muscle development in the limbs. AB - Homozygosity for the Splotch mutation causes neural tube and neural crest defects in mice. It has been demonstrated that Splotch mutant mice carry mutations in the homeodomain of the Pax-3 gene. Pax-3 is expressed in the neural tube, some neural crest derivatives, the mesenchyme of the limb bud and the somites. We have examined the development of the somite-derived skeletal muscles in homozygotes carrying the Splotch (Sp1H) mutation. Our results suggest that the Splotch mutation affects the development of skeletal muscles in a region-specific way: 1. The expression of the CMZ transgene in homozygotes reveals a disorganisation of the dermomyotome in whole stained embryos. 2. The axial musculature is reduced in size along a rostro-caudal gradient. 3. The muscle anlagen in the limbs develop much more slowly. Muscles of the head and the ventral body wall are normally developed in the mutant on day 13.5 of gestation. Recently, it has been shown that the myogenic precursors of the limbs are derived from the lateral half of the somite. The specific disturbance of muscle development in the limbs of Splotch mutants thus suggests a role for Pax-3 in the organisation of the somite, the production of trophic factors in the limb mesenchyme or an alteration of myogenic and mesenchymal cells. PMID- 8238964 TI - Adaptive remodelling of smooth muscle in the neo-intima of vein-to-artery grafts in rats: a detailed morphometric analysis. AB - End-to-end autogenous vein-to-artery grafts in rats have been used extensively as a model for neointimal thickening (hyperplasia), which develops over the first 6 weeks after grafting. This study employed computerised morphometric techniques to analyse 16 grafts, in order to quantitate precisely how the neointima develops. Two important features were described that have not been identified previously, due to the extensive variation in neo-intimal thickness inherent in vein grafts. Firstly, the proximal region of the graft was significantly thicker than the distal region, up until 6 months after grafting. The smooth muscle cells in the graft may have developed more rapidly in the proximal region, due to the altered haemodynamics within the graft. Secondly, within the central region of the graft the characteristic focal nature of neo-intimal hyperplasia was evident throughout the period of the study, but by 6 months the neo-intima tended to be distributed more evenly. By 6 months remodelling of smooth muscle throughout the graft neo intima had occurred, and the neo-intima had matured to a thickness equivalent to that of the intima plus media of the adjacent iliac artery. PMID- 8238966 TI - Ribosomes in the skeletal muscle filament lattice. AB - The compartmentalization of myosin isoforms within a muscle cell (Gauthier: J. Cell Biol. 110:693-701, 1990) suggests that myosin might be assembled directly into thick filaments at sites where it is synthesized. We therefore examined myofibrils by immunoelectron microscopy to determine whether ribosomes are associated with thick filaments under conditions in which new myosin can be identified. We used the embryonic chick anterior latissimus dorsi (ALD), a slow muscle that is induced, by curare, to synthesize a fast myosin isoform that is not normally present. Myosin was localized in situ, using a gold-labeled monoclonal antibody that recognizes the new isoform. The gold marker, as expected, was localized preferentially to the A band. There was an overall increase of fivefold in the number of gold particles per micron2 of A band in the curare-treated compared to the normal ALD, indicating that the labeled isoform was largely newly formed. There was a corresponding preferential distribution of ribosomes at the A band, especially in the H-band region, and the number of ribosomes per micron2 of A band was nearly twice as high in the curare-treated as in the normal muscle. Ribosomes were located between thick filaments, often aligned in rows. We conclude that ribosomes are located within the filament lattice, and therefore that they are available for local myosin synthesis. PMID- 8238965 TI - The primary visual system of flatfish: an evolutionary perspective. AB - The retinal projections of two species of flatfish (Scophthalmus maximus, Scophthalmidae; Platichthys flesus, Pleuronectidae) were investigated by autoradiography and by a HRP technique. Contralateral projections to five hypothalamic centres (area optica preoptica ventralis, nucleus opticus preopticus parvocellularis posterior pars lateralis, n. suprachiasmaticus, n. opticus hypothalami ventromedialis and area optica hypothalami posterior), thirteen thalamo-pretectal centres (nucleus opticus dorsolateralis (partes medialis, ventralis and lateralis), n. opticus ventrolateralis, n. opticus commissurae posterioris (partes dorsalis and ventralis), n. opticus accessorius, n. geniculatus lateralis mesencephali, nn. opticus pretectalis dorsalis, medialis and ventralis and n. corticalis), three layers of the optic tectum (stratum opticum pars externa, stratum fibrosum et griseum superficiale, stratum album centrale), and a single target in the tegmentum (n. opticus tegmenti mesencephali dorsalis), were identified in both species. Interspecific variation of the contralateral visual projections is relatively small. Ipsilateral visual projections of fibres which recross the midline in the minor and transverse commissures were also identified; in S. maximus this ipsilateral contingent is poorly developed and concerns principally hypothalamic structures, while in P. flesus the ipsilateral projections are considerably more extensive and involve both hypothalamic and thalamo-pretectal primary visual centres. No differences in the projections from the fixed and from the migrated eye were observed in either species. The findings are discussed in the general context of the existing literature on the visual projections of teleosts, in an attempt to characterize the primary visual system of the Pleuronectiformes in an evolutionary context. PMID- 8238967 TI - Giant mitochondria in the seromucous secretory cells of the accessory submandibular gland of the long-haired fruit bat, Stenonycteris lanosus. AB - Giant mitochondria, measuring up to 6.4 microns in diameter, are present in the seromucous secretory cells of the accessory submandibular gland of the long haired fruit bat, Stenonycteris lanosus. These mitochondria, as well as all of the smaller ones in the same cells, contain in their matrix compartment an abundance of 33 nm threads that probably consist of protein. Some mitochondria, regardless of size, contain 5 nm helical filaments within an expanded crista. Despite their altered morphology, the enlarged mitochondria in the accessory submandibular gland of S. lanosus must be able to function normally in energy metabolism, since the secretory cells in which they are found elaborate numerous secretory granules. PMID- 8238968 TI - Scanning and transmission electron microscopy and high resolution intravital video-microscopy of capillaries in the mouse exocrine pancreas, with special emphasis on endothelial cells. AB - Capillaries in the mouse exocrine pancreas were studied by scanning electron microscopy of microvascular corrosion casts, transmission electron microscopy of tissue sections, and high resolution intravital video-microscopy. Two types of capillaries were discerned by corrosion casting. The first type was rather straight, had a constant diameter of 5-6 microns, and its surface showed multiple circumferential furrows. The frequency of such constrictions was less in the second type, which was more undulated and had a diameter of 7-9 microns. In the second type, these constrictions defined bulged areas of the capillary cast. Corresponding tissue sections also showed two types of capillaries, fenestrated and non-fenestrated capillaries. Microtubules were abundant in all capillary endothelial cells, whereas bundles of microfilaments were scarce. Microtubules were arranged along the long axis of endothelial cells as well as parallel to endothelial cell border regions. Endothelial cells were joined by intermediate junctions along cell borders running both circumferentially and longitudinally. Flow reversal in capillaries and spontaneous endothelial contractions were documented in vivo. Endothelial cells bulged into the lumen, either at their nuclear region or distant from it. Spontaneous contraction of pericytes was not observed. These results suggest that contraction of capillaries is carried out by endothelial cells, representing an autonomous flow regulatory device. Capillary contraction in exocrine pancreas may be influenced by blood-borne agents, probably by those released in Langerhans islets. PMID- 8238969 TI - Innervation distribution pattern, nerve ending structure, and fiber types in pigeon skeletal muscle. AB - Four fiber types have been characterized in different pigeon skeletal muscles according to their innervation pattern (nerve ending structure and innervation distribution) and histochemical properties (SDH and m-ATPase activities). All fast fibers, types IIA and IIB, present aggregated distribution of their nerve endings with "en plaque" structures and very low innervation frequencies. The two kinds of slow fibers recognized are multiple innervated and present higher innervation frequencies. However, type I fibers have nerve terminals in small knobs with uniform localization, whereas type III fibers present "en grappe" nerve endings, which tend to be randomly distributed. Fiber type composition of skeletal muscles has been found closely related to their biomechanical function. Fast fibers are predominant in muscles with an active role in locomotive movements, whereas slow fibers are mainly or exclusively located in postural muscles. PMID- 8238970 TI - Sertoli cell cycle: a re-examination of the structural changes during the cycle of the seminiferous epithelium of the rat. AB - Seminiferous tubules in mammals are composed of cell associations that show a cyclic pattern of renewal and development. The cyclic nature of germ cell development suggests that the cells supporting the spermatogenic process, the Sertoli cells, might also differ structurally during the spermatogenic cycle in terms of the quantity of their constituents. In the present study, cyclic differences in volumes, and surface areas were determined using a sampling technique at the electron microscope level that proportionally samples the Sertoli cell within the seminiferous tubule. Among the many parameters studied, only the surface area of the cell, the volume of lipid, and the volume and surface area of the rough endoplasm reticulum were shown by statistical analysis to vary cyclically. Regarding rough endoplasm reticulum, the volume and surface area of this organelle peaked at mid-cycle and its low was recorded near the end of the cycle, exhibiting an approximate 15-fold difference between extremes. The rough endoplasm reticulum parameters generally correlated with known patterns of protein secretion within the tubule and with the secretion of specific proteins as well as the factors important in controlling protein secretion. Many Sertoli cell structural parameters suggested to be influenced cyclically in the rat in other studies could not be confirmed by the present study. Methodological differences in the present study and past studies are discussed as potential sources of error for these discrepancies. PMID- 8238971 TI - Ultrastructural changes in the efferent duct and epididymis of men with obstructive infertility. AB - Ultrastructural changes in the efferent duct and in different regions of the epididymis in men with obstructive azoospermia were compared with corresponding tissues collected from men of proven fertility who underwent castration due to malignancy of the prostate. Major degenerative changes were seen in the efferent duct and the caput epididymidis of men with obstruction at the caput epididymidis which may have been induced by fluid pressure due to defective absorption of testicular fluid in the caput epididymidis. These degenerative changes included decrease in tubular and lumen diameter of the caput and the cauda epididymides, decrease in height of the stereocilia, reduction in rough endoplasmic reticulum and Golgi material, and presence of lipofuscin and osmiophilic dense bodies. The degenerative changes were less when the site of obstruction was in the cauda epididymidis since fluid reabsorption would continue to take place normally in the caput epididymidis. In men who had undergone vasoepididymostomy (VEA), the ejaculated spermatozoa showed a high percentage of morphological abnormalities which may have occurred due to adverse effects of long-term obstruction on spermatogenesis. PMID- 8238972 TI - Ultra-high-resolution scanning electron microscopic studies on the membrane system of the parietal cells of the rat in the resting state and shortly after stimulation. AB - The three-dimensional organization of the membrane system of the rat parietal cells in the resting state and during early stimulation with tetragastrin (gastrin) was determined by ultra-high-resolution scanning electron microscopy. Specimens were prepared by cytoplasmic matrix removal using the aldehyde-osmium DMSO-osmium procedure. The intracellular canaliculus was lined with numerous microvilli. Viewed from the cytoplasmic side, the intracellular canaliculi appeared as an arborized system of cactus-like structures with numerous round holes about 100 nm in diameter corresponding to the basal openings of the microvilli. The intracellular canaliculi were more developed after gastrin stimulation than in the resting state. In resting cells, most of the tubulovesicles were isolated, 100-200 nm in diameter, spherical or tubular in shape, and had a smooth surface. After gastrin stimulation, these structures were interconnected by slender tubules of about 30 nm in diameter forming together tubulovesicular network. Occasionally, swollen and shrunken profiles were observed. The tubulovesicular network was connected with the intracellular canaliculus only at a few sites by the slender connecting tubules. Fusion of the tubulovesicular network with the intracellular canaliculus is observed at such sites. In the fasted rat, the microvilli were slender and their interior was packed with some kind of ill-defined material, probably microfilaments. However, after gastrin stimulation, the microvilli were swollen and their interior was almost empty. These morphological changes seem to indicate the accumulation of fluid in the microvilli after gastrin stimulation, with subsequent swelling. PMID- 8238973 TI - Ultrastructure of the glomus cells in the carotid body of chronically hypoxic rats: with special reference to the similarity of amphibian glomus cells. AB - The ultrastructural characteristics of the glomus cells in the rat carotid body exposed to extremely long-term hypoxia (10-12 weeks) were investigated. The glomus cells could be classified into four distinct types according to the shape of dense-cored vesicles in the glomus cell cytoplasm: 1) small vesicle cells (SVCs, 50 nm in mean diameter), 2) large vesicle cells (LVCs, 80 nm in mean diameter), 3) dilated eccentric vesicle cells (EVCs, 400-800 nm in diameter), and 4) mixed vesicle cells (MVCs, large and eccentric vesicles). Many clusters of glomus cells were found to contain all four categories of cell types. The appearance of EVCs was a unique and common characteristic of glomus cells in this long-term hypoxia model. We also noted other ultrastructural features with chronic hypoxia which are characteristic of the amphibian carotid labyrinth glomus cells: 1) incomplete covering of glomus cells with the supporting cell missing over a wide area, 2) long thin cytoplasmic projections in the intervascular stroma, and 3) intimate apposition of the glomus cells and pericytes (g-p connection), endothelial cells (g-e connection), plasma cells, and fibrocytes. Because arterial PO2 is generally low in amphibia, these may be general features of hypoxic adaptation and facilitate both uptake of oxygen from blood and release of catecholamine into the blood. The g-p and g-e connections may take part in the regulation of the microcirculation in the enlarged carotid body. PMID- 8238974 TI - Posterior pituitary of the newborn marsupial possum, Trichosurus vulpecula. AB - The fetal anterior pituitary-adrenal axis is thought to be involved in the initiation of birth in both eutherian and marsupial mammals. Little is known about the structure and function of the posterior pituitary at birth in the marsupial. Immunocytochemistry, high pressure liquid chromatography, and radioimmunoassay were used to identify vasopressin and mesotocin in the posterior pituitary of a newborn marsupial, the brushtail possum, Trichosurus vulpecula. The concentrations of vasopressin and mesotocin in the head of the newborn possum were 0.34 and 0.28 ng, respectively. The concentration of vasopressin was always greater than that of mesotocin, and the amounts of neuropeptides present in the head increased as the possum developed. PMID- 8238975 TI - Immune system of the spontaneously hypertensive rat: II. Morphology and function. AB - The spontaneously hypertensive rat (SHR) is a stress-sensitive animal which exhibits moderate immune dysfunction that has been implicated in the onset of hypertension. In this study, we examined the morphology of SHR thymus and spleen and further characterized the immune deficiency using Wistar-Kyoto (WKY) and Fisher 344 (F-344) rats for comparison. The adult SHR thymus does not display the increase in medullary volume typically noted with aging and the volume density of the marginal zone is decreased in the spleen. In vivo tritiated-thymidine incorporation is also decreased in the spleen of unstimulated SHR. In mixed lymphocyte reactions (MLR), the proliferative response of SHR splenocytes is significantly decreased relative to controls, WKY and F-344. Addition of interleukin-1 (IL-1), interleukin-2 (IL-2), or indomethacin to the MLR cultures does not increase proliferation. The proliferative response to T cell receptor monoclonal antibody (mAb-TCR) or interleukin-2 (IL-2) are similarly impaired in the SHR. The depressed proliferative T cell response is reversed by prolactin. It is suggested that the SHR is a valuable model for the study of immune deficiency. PMID- 8238976 TI - Effects of chronic fluoride exposure on morphometric parameters defining the stages of amelogenesis and ameloblast modulation in rat incisors. AB - The response of ameloblasts to long-term (6 weeks) exposure to 100 ppm fluoride was examined in continuously erupting mandibular incisors of female Sprague Dawley rats as compared to control rats receiving a similar diet (Teklad L-356) but no sodium fluoride in their drinking water. After treatment, animals from both groups were perfused intravascularly with glutaraldehyde, and the incisors were removed and processed for light microscope morphometric analyses directly from 1 microns thick Epon sections. Other animals were injected intravenously with calcein (green fluorescence) followed 4 hours later by xylenol orange (red fluorescence) in order to reveal smooth-ended ameloblast modulation bands and thereby allow quantification of parameters related to the creation and movement of modulation waves within the maturation zone of these teeth. The results indicated that rat incisors expressed four major changes in normal amelogenesis which could be attributed to the chronic fluoride treatment. First, ameloblasts produced a thinner than normal enamel layer by the time they completed the secretory stage and entered the maturation stage of amelogenesis. Second, enamel organ cells within the maturation zone, especially those from the papillary layer, were shorter in height than normal. Third, ameloblasts related to maturing enamel in areas where it was partially soluble and/or fully soluble in EDTA modulated at a rate that was much slower than normal. In some locations ameloblasts remained ruffle-ended for as much as 30% longer than normal per cycle. This upset the usual pattern such that fewer total modulation cycles were completed per unit time by these ameloblasts. Fourth, enamel proteins were lost from the maturing enamel layer at a rate that was about 40% slower than normal. The data suggested that ameloblasts detected the delay in the extracellular breakdown and/or loss of enamel proteins and they responded by remaining ruffle ended for longer intervals than usual (positive feedback). PMID- 8238977 TI - Development of the adult endocrine pancreas during metamorphosis in the sea lamprey, Petromyzon marinus L. I. Light microscopy and autoradiography. AB - The results of this study were found to support the hypothesis put forth by Boenig (Z. Mikrosk-Anat. Forsch., 17:125-184, 1929) that the caudal pancreas of adult lamprey develops from the epithelium of the extrahepatic common bile duct in that the bile duct cells were found to undergo a great proliferation during the early stages of metamorphosis, with a large number of the cells incorporating 3H-thymidine. If the bile duct degenerated as suggested by Barrington (The Biology of Lampreys. Academic Press, London, pp. 135-169, 1972), this uptake would not be expected. The cranial pancreas was determined to develop in a similar manner to the larval islets, with formation of the islets taking place within the intestinal/diverticular epithelium. The newly formed islets would migrate into the surrounding connective tissue. During the later stages of metamorphosis a small number of cells was found to incorporate the tritiated thymidine within mature islets. PMID- 8238978 TI - Development of the adult endocrine pancreas during metamorphosis in the sea lamprey, Petromyzon marinus L. II. Electron microscopy and immunocytochemistry. AB - The development of the adult endocrine pancreas was followed throughout metamorphosis in the sea lamprey using electron microscopy and immunocytochemistry. It was discovered that the caudal pancreas develops from the larval extrahepatic common bile duct through the process of transdifferentiation (dedifferentiation/redifferentiation). Early in metamorphosis the bile duct epithelial cells possess large vacuoles, resembling autophagic vacuoles, containing recognizable cell material. There is a loss of the large bundles of intermediate filaments characteristic of the larval bile duct epithelium. These same cells are then seen to contain granules immunoreactive for insulin. Pancreatic islets develop within the base of the bile duct epithelium from these transdifferentiated cells and migrate into the surrounding connective tissue to form the caudal pancreas. The cranial pancreas was found to develop from the epithelia lining the developing adult diverticulum and anterior intestine in a similar fashion as those in the larva. The second cell type to appear in either portion of the developing pancreas is similar to the third cell type of the adult: cells immunoreactive for somatostatin do not appear until late in metamorphosis in either region. PMID- 8238979 TI - Microdialysis of the pontine reticular formation reveals inhibition of acetylcholine release by morphine. AB - BACKGROUND: Systemically administered morphine inhibits rapid eye movement (REM) sleep; however, the neuronal mechanisms through which morphine disrupts REM sleep remain poorly understood. Recently, the authors have shown that morphine-mediated REM sleep inhibition is localized to a specific region of the pontine reticular formation: the gigantocellular tegmental field (FTG). Because cholinergic neurotransmission in the FTG is known to play a role in REM sleep generation, the present study examined the hypothesis that systemically administered morphine would cause decreased acetylcholine release in the FTG. METHODS: Microdialysis probes were stereotaxically positioned in the FTG of six barbiturate-anesthetized cats to measure acetylcholine release. Cholinergic input to the FTG arises from the laterodorsal (LDT) and pedunculopontine tegmental (PPT) brain stem nuclei. By electrically stimulating the LDT and PPT, it was possible to measure stimulation evoked acetylcholine release in the FTG. Morphine sulfate (MSO4) was administered intravenously (500 micrograms.kg-1). High performance liquid chromatography with electrochemical detection was used to measure stimulation-evoked acetylcholine release in the FTG before and after the systemic administration of morphine sulfate. RESULTS: Acetylcholine release in the pontine FTG was depressed significantly (P < 0.01) by systemic morphine sulfate. Acetylcholine release without electrical stimulation of the LDT and PPT averaged 0.6 +/- 0.18 (mean +/- SD) pmol/10 min of dialysis. Before morphine sulfate was administered, electrical stimulation of cholinergic LDT and PPT neurons increased acetylcholine release within the FTG to 1.9 +/- 0.76 pmol/10 min. After morphine sulfate was administered, there was a 37% decrease in acetylcholine release within the FTG to average values of 1.2 +/- 0.63 pmol/10 min. There was no significant effect of morphine sulfate on spontaneous acetylcholine release in the absence of LDT and PPT stimulation. CONCLUSIONS: Because FTG levels of acetylcholine release increase during REM sleep, the present results are consistent with the hypothesis that diminished acetylcholine release in the pontine FTG comprises one mechanism by which morphine inhibits REM sleep. PMID- 8238980 TI - Cerebral blood flow and metabolism in dogs with chronic diabetes. AB - BACKGROUND: Previously, the authors found that anesthetized diabetic dogs had increased cerebral blood flow (CBF) and oxygen consumption (CMRO2). These results may have been influenced by anesthesia or surgery. The aim of this study was to determine whether CBF and CMRO2 are increased in the awake or anesthetized state in the absence of acute surgical stress in diabetic dogs. A second aim was to determine whether increased CBF and CMRO2 in diabetic dogs are mediated through beta-adrenergic mechanisms. METHODS: Diabetic dogs (n = 8) underwent total surgical pancreatectomy followed by 4 months of insulin management (16 +/- 0.4 units/day, mean +/- SE) to maintain fasting and 3 PM blood glucose 10-17 mM. Control dogs (n = 8) underwent sham operation followed by a 4-month convalescence. Using previously inserted catheters, CBF (radiolabelled microspheres) and CMRO2 (sagittal sinus sampling) were measured before and after propranolol (2 mg/kg) in both the awake and anesthetized states. RESULTS: During the 4 months before CBF studies, the fasting blood glucose was greater in diabetic group than in the control group (11.0 +/- 0.3 vs. 4.0 +/- 0.1 mM, respectively). No difference occurred between groups in CBF or CMRO2. In the awake state, propranolol administration caused no CBF or CMRO2 changes. However, during anesthesia with 50 micrograms/kg fentanyl plus 10 mg/kg pentobarbital, propranolol administration decreased CBF in control, but not in diabetic, dogs. CONCLUSIONS: The authors' previous results showing increased CBF and CMRO2 with diabetes may be secondary to a differential response to acute surgical stress, a factor that was eliminated in this study. These results indicate that diabetes is associated with changes in the beta-adrenergic system that become evident under fentanyl/pentobarbital anesthesia. PMID- 8238981 TI - Barbiturate action is dependent on the conformational state of the acetylcholine receptor. AB - BACKGROUND: Barbiturates act on many neuronal ion channels by poorly understood mechanisms. The authors investigated the hypothesis that barbiturates inhibit the transient open-channel conformation of the nicotinic acetylcholine receptor (nAchR) by binding to a discrete site. METHODS: Inhibition curves of the agonist stimulated efflux of 8bRb+ from nAchR-rich membrane vesicles prepared from the electric tissue of Torpedo nobiliana were obtained for 14 barbiturates using a filter assay. RESULTS: When added simultaneously with agonist, all agents inhibited the ion efflux with half-inhibitory concentrations (IC50), varying from 23 microM for pentobarbital to 880 microM for barbital, and with Hill coefficients of one. The effect of several barbiturates on the agonist concentration-response curve for carbachol-stimulated efflux indicated that this inhibitory action was not competitive. CONCLUSIONS: The IC50s of these agents did not correlate with their octanol/water partition coefficients, nor with general anesthetic potency, although a degree of channel inhibition occurred with many agents at general anesthetic concentrations. The existence of a barbiturate inhibitory site of action was indicated by the structural specificity. This conclusion was supported by the Hill coefficient of one, and by the high inhibitory potencies, which ruled out membrane perturbations as a mechanism. This site on the transient open-channel conformation exhibits different structure activity relationships than an allosteric site established by equilibrium barbiturate binding on the resting conformation of the AchR. Thus, barbiturate action depends on the nAchR's conformational state. PMID- 8238982 TI - Effect of propofol infusion on splanchnic hemodynamics and liver oxygen consumption in the rat. A dose-response study. AB - BACKGROUND: Propofol has been used for the maintenance of anesthesia. The effects of propofol infusion on splanchnic hemodynamics and liver oxygen consumption, however, have not been reported. In the current investigation, the authors studied the effects of a continuous infusion of propofol on systemic and splanchnic hemodynamics using a new method to measure liver oxygen consumption in awake control and anesthetized rats. METHODS: Cannulas were inserted into the left ventricle, femoral artery, portal vein, and hepatic vein during ether anesthesia, and the rats were allowed to awaken and recover for 3-4 h before study. Animals were infused for 30 min with either saline (controls) or propofol at a rate of 300, 600, 900, or 1,200 micrograms.kg-1 x min-1. Cardiac output and organ blood flows were measured using radiolabelled microspheres, and blood samples from the femoral artery, portal vein, and hepatic vein were used to determine liver oxygen consumption. RESULTS: Mean arterial pressure decreased in a dose-dependent manner with a 25% reduction at the highest infusion rate. Systemic vascular resistance similarly decreased, whereas cardiac output remained unchanged at all the infusion rates. Hepatic arterial blood flow increased in a dose-dependent fashion over the dose range studied, to a maximum increase of 120%. Portal tributary blood flow increased by 30% at the highest infusion rate. Total liver blood flow increased in a dose-dependent manner to a maximum of 38%. Total oxygen delivery to the liver by the hepatic artery and portal vein increased in a dose-dependent fashion. Liver oxygen consumption increased in a dose-dependent fashion to a maximum increase of 51% at an infusion rate of 1,200 micrograms.kg-1 x min-1. The percent of oxygen extracted by the liver was not altered by propofol infusion, and hepatic venous oxygen saturation did not decrease at any dose studied. Coronary and renal blood flows were not altered. Arterial PaCO2, increased from 31 +/- 2 mmHg in awake control rats to 41 +/- 2 mmHg in spontaneously breathing rats infused with 1,200 micrograms.kg-1 x min-1 propofol. CONCLUSIONS: The maintenance of anesthesia using an infusion of propofol resulted in an increase in liver oxygen consumption that was fully compensated for by an increase in oxygen delivery to the liver. Splanchnic hemodynamics and liver oxygenation are not adversely affected during maintenance of anesthesia with propofol in the normal rat. PMID- 8238983 TI - Effect of halothane on hypoxic toxicity and glutathione status in cultured rat hepatocytes. AB - BACKGROUND: In hypoxic rats, halothane causes hepatotoxicity at oxygen levels that would cause minimal hepatotoxicity in the absence of halothane. Using a model that excludes systemic and extrahepatic effects of halothane, the authors tested the hypothesis that halothane hepatotoxicity in the whole-rat model is caused by a direct hepatotoxic effect of halothane, which is mediated by halothane-derived free radicals. METHODS: Rat hepatocyte monolayer cultures were exposed to defined gas phases for 2 h. Three experimental variables were present or absent: hypoxia (1% O2), halothane (2%), and cytochrome P-450 induction (by phenobarbital). Two experimental outcomes were measured: aspartate aminotransferase release, a measure of cell death, and reduced glutathione, an endogenous free radical scavenger whose levels are decreased by physiologically significant free radical injury. RESULTS: As anticipated, hypoxia increased cell death. Cytochrome P-450 induction by itself increased cell death during hypoxia. However, halothane had no effect on cell death during hypoxia, with or without cytochrome P-450 induction. Halothane had no toxic effect, even when glutathione was depleted before the onset of hypoxia. Glutathione was decreased moderately by hypoxia alone. Neither halothane nor cytochrome P-450 induction had any effect on glutathione levels. CONCLUSIONS: Halothane was not toxic, and it did not generate a physiologically significant free radical insult during hypoxia in the isolated rat hepatocyte under the experimental conditions used in testing. PMID- 8238984 TI - Activation of brain noradrenergic neurons during recovery from halothane anesthesia. Persistence of phasic activation after clonidine. AB - BACKGROUND: alpha 2-Adrenoceptor agonists, known as antihypertensive agents, may be used during general anesthesia for their anesthetic sparing action and to reduce the occurrence of side effects. Previous studies have shown that the brain's noradrenergic nucleus, locus coeruleus, is an important target in mediating the hypnotic action of alpha 2 agonists. The authors studied the effects of recovery from halothane anesthesia on the electrical activity of locus coeruleus neurons to examine cellular substrates underlying the clinical effectiveness of alpha 2 agonists. METHODS: Experiments were performed in locally anesthetized rats, whose circulatory and acid-base stabilities were ensured by mechanical ventilation and volume infusion. Locus coeruleus neurons were recorded continuously while the rats were anesthetized with halothane (1%) and/or after the halothane was discontinued. RESULTS: Under the influence of halothane, locus coeruleus cells exhibited a slow, regular spontaneous discharge (1.95 +/- 0.23 Hz), and contralateral foot or tail pinch elicited a prominent, phasic activation in locus coeruleus neurons. Such phasic activation was blocked by local ejection of kynurenic acid, an excitatory amino acid antagonist, close to recorded neurons, but not by clonidine (up to 64 micrograms.kg-1). Thirty minutes after the halothane was discontinued, the mean firing rate of locus coeruleus neurons was increased by 87 +/- 20%. This excitation resulted from a prominent increase in bursting activity (21 +/- 5% of spikes in bursts vs. 4 +/- 1%) and was reversed by halothane readministration. This activation also was reduced by local ejection of kynurenic acid. Halothane discontinuance revealed the reactivity of locus coeruleus neurons to nonnoxious, sensory stimuli, and considerably reduced the apparent potency of intravenous administration of clonidine to inhibit locus coeruleus activity (effective dose for 50% of maximal effect (ED50), 25.48 +/- 8.26 micrograms.kg-1 vs. 4.81 +/- 0.80 micrograms.kg-1 under halothane). This decrease was caused by the persistence of bursting activity after the administration of clonidine, which was completely suppressed by readministration of halothane or local application of kynurenic acid. CONCLUSION: The data demonstrate: (1) that halothane withdrawal increases locus coeruleus neuronal activity via excitatory amino acid input, and this withdrawal-induced activity is characterized by a prominent burst (phasic) discharge; (2) that sedative doses of clonidine inhibit the tonic component of locus coeruleus activity but not the phasic activation of locus coeruleus neurons; and (3) that readministration of halothane or local ejection of an excitatory amino acid antagonist fully suppresses the bursting activity unaffected by clonidine. PMID- 8238985 TI - Intratracheal perfluorocarbon administration as an aid in the ventilatory management of respiratory distress syndrome. AB - BACKGROUND: Respiratory distress syndrome carries a high morbidity and mortality when treated with mechanical ventilation with positive end-expiratory pressure. Perfluorocarbon liquids are employed in liquid ventilation due to low surface tension and high gas solubility. To assess whether intratracheal administration of the perfluorocarbon, perflubron, in combination with conventional mechanical ventilation could be of therapeutic benefit in respiratory distress syndrome, the authors tested the effects of different doses of intratracheal perflubron administration on gas exchange and lung mechanics in adult animals with respiratory failure during a 6-h observation period. METHODS: Respiratory failure was induced in 30 rabbits by saline lung lavage (arterial oxygen tension < 100 mmHg at 100% oxygen with the following ventilator settings: tidal volume, 12 ml.kg-1; respiratory frequency, 30 per min; inspiratory/expiratory ratio, 1:2; and positive end-expiratory pressure of 6 cm H2O). Twenty-four rabbits were treated with different perfluorocarbon doses (3, 6, 9, and 12 ml.kg-1), and the remaining six served as controls while mechanical ventilation was continued with the aforementioned settings. Additionally, in ten healthy rabbits who were used as healthy controls, the lungs were mechanically ventilated either alone or in combination with intratracheal perfluorocarbon administration (3 ml.kg-1) for 6 h. RESULTS: In all treatment groups, arterial oxygen pressure increased significantly (P < 0.0001) in a dose-related fashion (193 +/- 40, 320 +/- 70, 353 +/- 125, and 410 +/- 45 mmHg at 15 min), and peak airway pressures decreased significantly (range, 18-23%; P < 0.0001) from pretreatment values. These findings were in contrast to those for the control group. The improvements were time-dependent in all four tested perfluorocarbon doses. However, the improvements in pulmonary parameters could be extended to 6 h only in groups treated with 9 ml.kg-1 and 12 ml.kg-1 perflubron. At the end of the 6-h period, the data for these two groups showed significantly higher arterial oxygen pressure (230 +/- 84 and 197 +/- 130 mmHg, respectively; P < 0.05) and lower inflation pressures than the pretreatment data for these groups and the data for the control group at 6 h. There were no clinically significant changes in pulmonary parameters in healthy animals due either to mechanical ventilation alone or mechanical ventilation in combination with intratracheal perfluorocarbon administration for 6 h. CONCLUSIONS: The results of this study imply that there is no association between the lung mechanics and gas exchange parameters for mechanical ventilation in combination with intratracheal perfluorocarbon administration. The data suggest that this type of perfluorocarbon administration with conventional mechanical ventilation offers a simple, alternative treatment of respiratory distress syndrome. With this technique, adequate pulmonary gas exchange can be maintained at relatively low airway pressures with high perfluorocarbon doses for several hours. PMID- 8238986 TI - Tolerance to sodium nitroprusside. Studies in cultured porcine vascular smooth muscle cells. AB - BACKGROUND: Tolerance to nitrovasodilators, including sodium nitroprusside (SNP), is a clinical problem. SNP causes vasodilatation by releasing nitric oxide, which stimulates intracellular guanosine 3':5'-cyclic monophosphate (cGMP) accumulation in smooth muscle cells. This study examined tolerance to SNP in coronary smooth muscle cells by measuring intracellular cGMP formation. METHODS: Smooth muscle cells were isolated from pig coronary vessels. Intracellular cGMP formation in response to SNP, nitroglycerine, and atrial natriuretic peptide (ANP) was quantitated by radioimmunoassay before and after chronic pretreatment. RESULTS: The detection of tolerance of cells rechallenged with SNP required exposure to a minimum concentration of 1 microM for SNP for at least 60 min. In untreated cells, acutely applied 100 microM SNP increased the cGMP concentration to 300 +/- 8.6 pmol/mg protein (mean +/- SEM). The cGMP concentration in cells pretreated with 1 microM SNP for 60 min increased to 213 +/- 11 pmol/mg in response to 100 microM SNP (P < 0.01, compared to untreated cells). The pretreatment of cells for 60 min with increasing concentrations of SNP caused a sequential reduction in the acute concentration-response curves of cGMP to SNP. After a 6 h washout period, cGMP accumulation in response to a rechallenge with SNP returned to control levels. The acute concentration-response curve of cGMP to SNP in cells pretreated with 100 microM nitroglycerine for 60 min was reduced significantly. Pretreatment with 100 microM SNP for 60 min did not change the response of cAMP accumulation to isoproterenol or forskolin. However, the response of cGMP formation to ANP was potentiated after pretreatment. In addition, elevation of basal cAMP or cGMP concentrations, induced with isoproterenol or ANP, respectively, did not alter the response of cGMP formation to SNP. CONCLUSION: The study yielded the following findings: (1) reversible tolerance to SNP was induced in cultured vascular smooth muscle cells; (2) chronic exposure to nitroglycerine induced cross-tolerance to SNP; (3) cells that were tolerant to SNP showed a potentiated response to ANP. PMID- 8238987 TI - Left ventricular diastolic function in the normal and diseased heart. Perspectives for the anesthesiologist (2). AB - Several important questions remain to be answered by future research. First, it is unclear whether any abnormal index of diastolic function can be used to estimate disease severity, or to prognostically identify patients who will subsequently develop systolic abnormalities or frank left ventricular dysfunction. A temporal relationship between the appearance of diastolic dysfunction and ultimate left ventricular decompensation may, theoretically, exist, but such a relationship has yet to be established. Second, a growing body of evidence indicates that pharmacologic therapy with Ca2+ channel antagonists, beta-adrenergic agonists or antagonists, phosphodiesterase inhibitors, or angiotensin converting enzyme inhibitors may acutely or chronically benefit certain patients with diastolic dysfunction. Whether the impact of early recognition and therapeutic intervention in patients with diastolic dysfunction can be translated into an improvement of quality of life or enhanced survival remains unknown. Third, recent evidence indicates that fundamental changes in the biochemistry of the cardiac myocyte may represent a final common pathway for the development of congestive heart failure resulting from intrinsic cardiac disease. Altered expression of genes coding for the ATP-dependent Ca2+ pumps in the sarcolemma and the sarcoplasmic reticulum, regulatory proteins such as phospholamban, and the proteins composing the contractile apparatus have been identified that play critical roles in the pathophysiology of myocardial failure, and have important implications for potential pharmacologic therapy. Future research will more clearly elucidate these cellular and biochemical mechanisms of left ventricular failure. Lastly, although intravenous and inhalational anesthetics produce derangements in normal diastolic function to varying degrees, whether the effects of these agents on diastolic performance are exacerbated in disease processes manifested by abnormal diastolic mechanisms requires further evaluation. PMID- 8238988 TI - Ventricular tachycardia and brief cardiovascular collapse in two infants after caudal anesthesia using a bupivacaine-epinephrine solution. PMID- 8238989 TI - Operating room fires initiated by hot wire cautery. PMID- 8238990 TI - Respiratory and circulatory compromise associated with acute hydrothorax during operative hysteroscopy. PMID- 8238991 TI - Intraoperative bronchospasm caused by adenosine. PMID- 8238992 TI - Intravenous abuse of transdermal fentanyl therapy in a chronic pain patient. PMID- 8238993 TI - Neurologic changes during carotid endarterectomy. PMID- 8238994 TI - Relationships with industry are ok if you follow the rules. PMID- 8238995 TI - Fiberoptic intubation and the laryngeal mask airway. PMID- 8238996 TI - Parental screening before intraoperative magnetic resonance imaging of a child. PMID- 8238997 TI - Topical anesthesia of the airway using the lighted stylet. PMID- 8238998 TI - Postoperative pulmonary complications: I. PMID- 8238999 TI - Postoperative pulmonary complications: II. PMID- 8239000 TI - Spurious anesthesia alarm in an anesthetized patient. PMID- 8239001 TI - Laryngeal mask airway in the cannot-intubate, cannot-ventilate situation. PMID- 8239002 TI - Latex anaphylaxis masquerading as fentanyl anaphylaxis: retraction of a case report. PMID- 8239003 TI - Catheter location and patient position affect spread of interpleural regional analgesia. PMID- 8239004 TI - Endobronchial intubation by a nonbeveled endotracheal tube in infants and small children. PMID- 8239005 TI - Anterior spinal artery syndrome? PMID- 8239006 TI - Association of University Anesthesiologists satellite symposium: the role of the endothelium in vascular control. Asheville, North Carolina, May 2-4, 1993. PMID- 8239007 TI - Brain protection during anesthesia. A reader's guide. PMID- 8239008 TI - Mechanisms of incomplete cardioplegia distribution during coronary artery surgery. An intraoperative transesophageal contrast echocardiography study. AB - BACKGROUND: Cardioplegia is used to protect the myocardium from ischemic injury during open-heart surgery. However, the delivery of cardioplegic solutions may be impaired by anatomic and/or functional conditions, such as the development of transient aortic regurgitation during antegrade administration of cardioplegia or shunting through a foramen ovale during retrograde administration. In this study, the authors used a new method of cardioplegia administration, based on intraoperative contrast echocardiography, to detect on-line causes of inadequate cardioplegia delivery. METHODS: Forty patients with coronary artery disease and a competent aortic valve, who were treated consecutively, were enrolled in this study. Patients were monitored intraoperatively by transesophageal contrast echocardiography during cardioplegia delivery. Antegrade cardioplegia was administered into the aortic root following aortic occlusion in all patients. Twenty-two patients also received retrograde cardioplegia, administered through the right atrium. The echo-contrast agent consisted of a stable suspension of 5% human albumin microbubbles with a concentration of 4 x 10(8) microbubbles/ml and a diameter of 4 +/- 1 mu. RESULTS: Antegrade cardioplegia was not associated with aortic regurgitation in 23 of 40 (58%) patients. Seven patients (17%) had only mild aortic regurgitation, four patients (10%) had moderate regurgitation, and six (15%) had severe aortic regurgitation. The percent of myocardial opacification was 76.0 +/- 10.5 in the 23 patients who did not have aortic regurgitation, 76.0 +/- 17.0 in the 7 patients who had mild regurgitation, 52.5 +/- 18.1 in the 4 patients who had moderate regurgitation, and 48.5 +/- 18.3 in 6 patients who had severe aortic regurgitation (Kruskal-Wallis stat, 12.9; P < 0.005). Retrograde cardioplegia was not associated with right-to-left shunt in 11 of 22 patients (50%). In seven patients (32%), there was only a mild passage of contrast material to the left atrium. In the remaining four patients (18%), there was a moderate (one patient) to severe (three patients) right-to-left shunt at the level of the fossa ovalis. CONCLUSIONS: This study shows that incomplete myocardial distribution of cardioplegia, secondary to transient aortic valve incompetence or shunting through the foramen ovale, is not uncommon in patients undergoing coronary surgery. PMID- 8239009 TI - Pharmacodynamic and hemodynamic effects of mivacurium in infants anesthetized with halothane and nitrous oxide. AB - BACKGROUND: The newly developed neuromuscular blocking agent, mivacurium, has been evaluated in adults and children, but there are no data on its effects in infants. This study was designed to evaluate the neuromuscular effects of mivacurium by dose-response analysis, and its cardiovascular effects in 90 infants 2-11 months of age anesthetized with 1% halothane and nitrous oxide:oxygen. METHODS: The neuromuscular response was measured by recording the force of contraction of the adductor pollicis during train-of-four stimulation at 0.1 Hz. The infants were divided according to age into two equal groups of 45; group A infants were 2-6 months of age, and group B infants were 7-11 months of age. Each group was further subdivided into five subgroups of nine. Infants in group A received mivacurium at sequential doses of 40, 50, 55, 75, and 150 micrograms/kg, while those in group B received mivacurium at doses 40, 50, 60, 75, and 150 micrograms/kg. The first four doses in each group were used to determine dose-response relationships. The last two doses of 75 and 150 micrograms/kg were based on the observed preceding dose-response data to approximate the ED95 and 2XED95. Heart rate and blood pressure were determined every minute for a minimum of 3 min after mivacurium. RESULTS: The effective doses for 50% depression of the first twitch response of the train-of-four (ED50) were 44-50 micrograms/kg (confidence limits 29-74 micrograms/kg), without any significant difference between groups A and B. In both groups, a larger dose of mivacurium, 150 micrograms/kg, caused complete ablation of the twitch response in 1.3 +/- 0.2 min (mean +/- SE) with recovery to 5, 25, and 95% of control in 7.6 +/- 0.5, 9.4 +/- 0.6, and 16.2 +/- 0.9 min, respectively. In infants, the 25-75% recovery index was 3.8 +/- 0.4 min, and the 5-95% recovery index was 8.5 +/- 0.8 min. In 28 infants, in whom surgical relaxation was required for more than 20 min, the infusion requirements to maintain 90-99% neuromuscular block in infants 2-6 and 7-11 months of age were 12.1 +/- 1 and 9.9 +/- 1 micrograms.kg-1.min-1, respectively (NS). No significant changes of heart rate of blood pressure occurred in infants, except in the subgroup of infants 7-11 months of age who received 150 micrograms/kg mivacurium. In this group, a 13-mmHg increase in mean systolic blood pressure was seen without any significant change in diastolic pressure or heart rate. In addition, in 7 of 36 patients receiving 75-150 micrograms/kg mivacurium, a greater than 29% change in systolic or diastolic pressure occurred. One infant with cholinesterase deficiency had a prolonged neuromuscular block from mivacurium. CONCLUSIONS: The ED50 duration of action and infusion requirements of mivacurium in infants 2-6 months of age are comparable with those of infants 7-11 months of age. PMID- 8239010 TI - Efficacy of oral clonidine premedication in children. AB - BACKGROUND: Clonidine, an alpha 2-adrenoceptor agonist, has been shown to be effective as a preanesthetic medication in adults. The current study was designed to investigate the efficacy of two doses of oral clonidine as a premedicant preceding oral atropine in children. METHODS: In a prospective, randomized, double-blind, controlled clinical trial, 105 children, aged 4-12 yr, undergoing elective ophthalmologic surgery received 0.4 mg/kg diazepam, 2 micrograms/kg clonidine, or 4 micrograms/kg clonidine orally. These agents mixed with apple juice were administered 105 min before the estimated time of induction of anesthesia, and were followed by treatment with 0.03 mg/kg oral atropine 60 min before anesthesia. A blinded observer noted the children's level of sedation, quality of separation from parents, and degree of acceptance of mask application during inhalation of nitrous oxide used for establishment of venous access. Anesthesia was induced with 5 mg/kg thiamylal, and tracheal intubation was facilitated with 0.2 mg/kg vecuronium. Hemodynamic changes after tracheal intubation were compared among the three groups. RESULTS: Clonidine produced significant sedation, and the effect was dose related. Clonidine, 4 micrograms/kg, provided better quality of separation and acceptance of mask than the two other regimens. This dose of clonidine attenuated the increases in blood pressure and heart rate after tracheal intubation. No clinically significant perioperative hypotension or bradycardia was observed. CONCLUSIONS: These data indicate that, even in pediatric surgery, the combination of 4 micrograms/kg and 0.03 mg/kg oral clonidine is an effective premedication. However, the safety and optimal dose of clonidine in this setting remain to be determined. PMID- 8239011 TI - The predictive performance of a system model for enflurane closed-circuit inhalational anesthesia. AB - BACKGROUND: Previously, the authors described a system model for closed-circuit inhalational anesthesia, and demonstrated close agreement between end-tidal isoflurane concentrations measured in their clinical study and those predicted by the model. The predictive performance of their model has not, however, been tested for anesthetics featuring nonpulmonary elimination (NPE). METHODS: The authors quantified the predictive performance of two versions (A and C) of the model in 50 patients by comparing the predicted and the measured alveolar concentration-time profiles after bolus injections of liquid enflurane into the expiratory limb of the closed system. Version A did not incorporate NPE, but version C emulated NPE by adopting the irreversible loss of a fraction of the enflurane present in the arterial hepatic blood flow (0.131, derived from a mass balance study performed by others). For each concentration measured by mass spectrometry, the authors used computer simulations of version A and C to calculate a predicted concentration for both versions. For each patient, the authors calculated the bias (indicating systematic over- or underprediction) and the scatter of the prediction errors (indicating typical error size). RESULTS: The authors administered a total of 379 ml of liquid enflurane via 466 injections. A total of 18,432 alveolar concentrations (one per 10-s period; average concentration = 0.96 vol%) were measured. The bias and the scatter, both given as mean (and SD), were 10.0 (13.1)% and 11.8 (3.9)% for version A and -0.8 (11.4)% and 11.4 (2.8)% for C. The bias for version C was closer to zero; the scatters were similar. CONCLUSIONS: Version C incorporating NPE performs better than version A. The accuracy that was obtained should encourage the use of version C for clinical, teaching, research, economic, and ecologic purposes. PMID- 8239012 TI - Succinylcholine increases intraocular pressure in the human eye with the extraocular muscles detached. AB - BACKGROUND: The increase in intraocular pressure in the human eye that is associated with the use of intravenous succinylcholine has long been ascribed to contraction of the extraocular muscles leading to compression of the globe. This created concern that such contraction would extrude global contents in the patient with an open globe, and led clinicians to avoid the use of succinylcholine in these patients. METHODS: The authors studied 15 patients undergoing elective enucleation, and compared the intraocular pressure change after the administration of succinylcholine in the diseased eye after all the extraocular muscles had been detached to that of the normal eye that had the extraocular muscles attached. RESULTS: The authors found no difference in baseline intraocular pressure (mmHg) between eyes (15.1 vs. 16.1) or at peak intraocular pressure (25.2 vs. 24.7), but did observe a significant difference in pressure in both eyes when baseline was compared with peak pressure. CONCLUSIONS: The authors concluded that extraocular muscle contraction does not contribute to the increase in intraocular pressure after succinylcholine. PMID- 8239013 TI - The dose of propofol required to prevent children from moving during magnetic resonance imaging. AB - BACKGROUND: Intravenous propofol offers several advantages as an anesthetic for children undergoing magnetic resonance imaging. However, the dose of propofol required to prevent movement during magnetic resonance imaging is likely to be less than that required for surgical anesthesia. METHODS: Thirty children between the ages of 1 and 10 years, undergoing elective magnetic resonance imaging as outpatients were randomly assigned to receive a propofol infusion at a rate of 50, 75, or 100 micrograms.kg-1.min-1 during the imaging procedure. Anesthesia was induced with inhalation of halothane, nitrous oxide, and oxygen, and a 2 mg.kg-1 loading dose of propofol. Immediately after insertion of an intravenous catheter, inhaled anesthetics were discontinued and the propofol infusion started. The children then were observed for movement during the scan. RESULTS: There were no significant differences among the three groups with respects to mean age (4.4 +/- 2.0 yr), weight (17.6 +/- 5.1 kg), induction time (11 +/- 3 min), scan duration (55 +/- 26 min), or recovery time (30 +/- 8 min). Five of ten patients who received 50 micrograms.kg-1 x min-1 moved during the scan, three of ten patients who received 75 micrograms.kg-1 x min-1 moved, and none of the children who received 100 micrograms.kg-1 x min-1 moved. Two patients experienced a decrease of arterial oxygen saturation to less than 95% after receiving the initial bolus of propofol. The arterial oxygen saturation returned to normal within 15 s without specific treatment other than continued supplemental oxygen. There were no episodes of hypoxemia during image acquisition. None of the children experienced nausea or vomiting. CONCLUSIONS: Following induction of anesthesia with halothane, nitrous oxide, and a 2 mg.kg-1 loading dose of propofol, infusion of propofol at a rate of 100 micrograms.kg-1 x min-1 effectively prevents children from moving during elective magnetic resonance imaging. A transient decrease in arterial oxygen saturation can occur after the initial bolus of propofol. Recovery from anesthesia is rapid and without nausea or vomiting. PMID- 8239014 TI - Isoflurane minimum alveolar concentration decreases during anesthesia and surgery. AB - BACKGROUND: It generally is assumed that the potency of inhalational anesthetics remains unchanged during the course of the administration of an anesthetic. Only one study has indicated a decrease of minimum alveolar concentration with time. In this study, an effect of the duration of anesthesia administration and surgery on the potency of isoflurane was investigated by determining MACtetanus (the minimum alveolar concentration that prevents movement in response to electrical tetanic stimulation in 50% of patients) before and after surgery. METHODS: Ten patients who underwent removal of a herniated intervertebral disc were anesthetized with isoflurane only. Reaction to a standardized electrical stimulation applied to the forearm was observed and was graded as movement or no movement. The isoflurane concentration was increased in steps of 0.10 vol% if the patient moved and decreased in steps of 0.10 vol% if no reaction was seen, until a "movement/no-movement/movement" or "no-movement/movement/no-movement" pattern, respectively, was achieved. RESULTS: MACtetanus decreased in all patients from 1.28 +/- 0.22 vol% (mean +/- SD) before surgery to 1.04 +/- 0.22 vol% after surgery (P < 0.01). When the prestimulation arterial blood pressure or the maximal increase in blood pressure caused by stimulation at the individual MACtetanus before surgery were compared to the corresponding values at the individual MACtetanus after surgery, no significant difference could be found. The prestimulation heart rate and the maximal increase in heart rate were significantly lower after surgery, even though the end-tidal isoflurane concentration was 0.24 vol% lower at the individual MACtetanus after surgery. CONCLUSION: The authors conclude that MACtetanus decreases during the administration of anesthesia and the performance of surgery. PMID- 8239015 TI - Diaphragmatic function before and after laparoscopic cholecystectomy. AB - BACKGROUND: Diaphragm dysfunction is a primary cause of ventilatory impairment after upper abdominal surgery. Laparoscopic procedures may result in less dysfunction. To test this, diaphragmatic function was studied in ten healthy adult patients undergoing elective laparoscopic cholecystectomy and in five undergoing laparoscopic hernia repair. METHODS: Respiratory gas exchange, ventilation, and breathing pattern were measured before and 3 h after surgery. Respiratory drive was evaluated from the relationship of P0.1 to end-tidal carbon dioxide (PETCO2) during tidal breathing. Diaphragm contractile function was assessed from maximal transdiaphragmatic pressure (Pdimax), and Pdi during a maximal sniff maneuver (Pdisniff). RESULTS: Oxygen consumption and carbon dioxide production did not change after surgery. Pdimax decreased by more than 50% in the laparoscopic cholecystectomy group, but Pdisniff did not change. Tidal volume and the ratio of inspiratory time over total cycle time decreased by 30% and 13%, respectively, PETCO2 increased by 9%, and minute ventilation did not change. In contrast, there was no variation in ventilatory function in patients undergoing laparoscopic hernia repair. In both groups, P0.1 did not change, which excludes depressed respiratory drive as an explanation for the decreased Pdimax in laparoscopic cholecystectomy. Contractile failure of the diaphragm was discounted as well, because Pdisniff did not change, even in the laparoscopic cholecystectomy group. CONCLUSIONS: Although laparoscopic cholecystectomy does not increase metabolic demands in the early postoperative period, it impairs diaphragm function. The internal site of surgical intervention appears to be the critical variable determining diaphragmatic inhibition after laparoscopic abdominal surgery. PMID- 8239016 TI - Thoracic intravascular and extravascular fluid volumes in cardiac surgical patients. AB - BACKGROUND: One possible mechanism of impaired oxygenation in cardiac surgery with extracorporeal circulation (ECC) is the accumulation of extravascular lung water (EVLW). Intrathoracic blood volume (ITBV) and pulmonary blood volume (PBV) also may increase after separation from ECC, which can influence both cardiac performance and pulmonary capillary fluid filtration. This study tested whether there were any relationships between lung fluid accumulation and pulmonary gas exchange during the perioperative period of cardiac surgery and ECC. METHODS: Ten patients undergoing myocardial revascularization were studied. ITBV, PBV, and EVLW were determined from the mean transit times and decay times of the dye and thermal indicator curves obtained simultaneously in the descending aorta. Gas exchange was assessed by arterial and mixed venous partial pressure of oxygen (PO2) and carbon dioxide (PCO2), and calculation of alveolo-arterial PO2 gradient (PA-aO2) and venous admixture (QVA/QT). Recordings were made after induction of anesthesia, after sternotomy, 15 min after separation from ECC, and 4 and 20 h postoperatively. RESULTS: After induction of anesthesia, EVLW (6.0 +/- 1.0 ml/kg, mean +/- SD), PBV (3.6 +/- 1.3 ml/kg), and ITBV (18.4 +/- 2.7 ml/kg) were within normal ranges. Oxygenation was moderately impaired, as indicated by an increased PA-aO2 (144 +/- 46 mmHg) and QVA/QT (11 +/- 4%). After separation from ECC, EVLW had increased to 9.1 +/- 2.6 ml/kg, which was accompanied by an increase of ITBV (26.0 +/- 4.4 ml/kg) and PBV (5.6 +/- 1.9 ml/kg). PAa-O2 (396 +/- 116 mmHg) and QVA/QT (29 +/- 7%) also were increased. ITBV and PBV remained increased 4 and 20 h postoperatively, but EVLW decreased to presurgery values. No correlations were found between thoracic intravascular and extravascular fluid volumes and gas exchange. CONCLUSIONS: Cardiac surgery with the use of ECC induces alterations of thoracic intravascular and extravascular fluid volumes. Postoperatively, increased ITBV and PBV need not be associated with higher EVLW. Thus, sufficient mechanisms protecting against lung edema formation or providing resolution of EVLW probably are maintained after ECC. Since oxygenation is impaired during and after cardiac surgery, it is concluded that mechanisms other than or in addition to changes of ITBV, PBV, and EVLW predominantly influence gas exchange. PMID- 8239017 TI - Sevoflurane and halothane reduce focal ischemic brain damage in the rat. Possible influence on thermoregulation. AB - BACKGROUND: There has been little systematic examination concerning the comparative effects of the anesthetized versus the awake state on outcome from cerebral ischemia. This experiment evaluated infarct volume and neurologic function in rats subjected to temporary focal ischemia while anesthetized with either sevoflurane or halothane. Outcome in these animals was compared to that observed in rats maintained unanesthetized during a similar ischemic insult. METHODS: All rats were anesthetized with halothane and surgically prepared for filament occlusion of the middle cerebral artery. After preparation, one group (Halothane) remained anesthetized with approximately 1.4 MAC halothane. In another group (Sevoflurane), halothane was discontinued and substituted with sevoflurane, which was administered until electroencephalographic burst suppression was evident (approximately 1.4 MAC). The final group (Awake) was allowed to awaken immediately after the onset of ischemia. Middle cerebral artery occlusion persisted for 90 min in all groups. The middle cerebral artery filament then was removed, and a 96-h survival interval was allowed. Neurologic function and infarct volume were determined. Recent evidence indicates that transient mild hyperthermia occurs in awake rats undergoing filament occlusion of the middle cerebral artery. To examine the potential role of mild hyperthermia in this experiment, a second experiment was performed in which rats anesthetized with halothane underwent 90-min focal ischemia, with pericranial temperatures held at either 38.0 degrees C or 39.2 degrees C. RESULTS: Intraischemic mean arterial pressure was 20-25 mmHg lower in the two anesthetized groups compared with awake animals. Despite this finding, cortical infarct volumes (mean +/- SD; Halothane, 17 +/- 32 mm3; Sevoflurane, 36 +/- 57 mm3; Awake, 115 +/- 104 mm3; Sevoflurane, 36 +/- 57 mm3; Awake, 115 +/- 104 mm3) and subcortical infarct volumes (mean +/- SD; Halothane, 39 +/- 57 mm3; Sevoflurane, 50 +/- 29 mm3; Awake, 88 +/- 46 mm3) were reduced in both groups of anesthetized rats. This reduction correlated with improved neurologic function. The rats in whom the pericranial temperature was maintained at 39.2 degrees C had a larger total infarct volume (218 +/- 81 mm3) and increased neurologic deficits when compared to those in whom the pericranial temperature was maintained at 38.0 degrees C (total infarct volume, 75 +/- 77 mm3). CONCLUSIONS: Both halothane and sevoflurane substantially reduced damage in this focal ischemia model when compared to outcome resulting from the same insult induced in awake rats. The reduction in intraischemic mean arterial pressure caused by the anesthetics did not seem contributory to outcome. Brain temperature differences among the groups were not defined. Because small differences in pericranial temperature were shown to have major effects on outcome, further work is required to determine if differences in brain temperature explain the observed protective effects of these anesthetics. PMID- 8239018 TI - Role of the injury discharge in the development of thermal hyperesthesia after sciatic nerve constriction injury in the rat. AB - BACKGROUND: Usually, a barrage of impulses ("injury discharge") is evoked following sensory nerve damage. It has been suggested that injury discharge may produce the hyperexcitatory state in the spinal cord, and this hyperexcitability may cause neurogenic pain. In the present study, the authors examined the role of injury discharge in developing the hyperesthetic state following nerve constriction injury. METHODS: A model of thermal hyperesthesia caused by a constriction injury created by making four loose ligations around the rat sciatic nerve was examined. To block the injury discharge, 0.5% bupivacaine was applied to the sciatic nerve before constriction injury. To block the hyperexcitatory state, (+)-MK-801, an N-methyl-D-aspartate antagonist, was administered intrathecally 15 min before the nerve lesion. RESULTS: Blocking injury discharge significantly delayed the development of hyperesthesia. Bupivacaine had no effect on the development of hyperesthesia when bupivacaine was applied to the sciatic nerve 15 min after the nerve constriction injury. Systemic bupivacaine had no effect on the development of thermal hyperesthesia. Intrathecal (+)-MK-801 also delayed the development of hyperesthesia when (+)-MK-801 was administered intrathecally 15 min before the nerve injury. When (+)-MK-801 was administered 15 min after the nerve injury, (+)-MK-801 had no effect on the development of hyperesthesia. CONCLUSION: These results suggest that injury discharge may induce facilitation of spinal dorsal horn neurons, and this spinal facilitation may play an important role in developing thermal hyperesthesia following sciatic nerve constriction injury. PMID- 8239019 TI - [The effect of diprivan on the indices of central hemodynamics and the heart conduction system]. AB - Using impedance tetrapolar plethysmography and transesophageal electrical stimulation of the left ventricle, changes in central hemodynamic parameters have been assessed in 46 women during brief (up to 10 min) anesthesias with diprivanum. Bolus injection of diprivanum at a dose of 2.5 mg per 1 kg body weight was used. It has been demonstrated that diprivanum administration decreased BP, which was associated with a drop in total peripheral resistance and stroke volume that led to the absence of compensatory tachycardia to an increase in the cardiac output. The absence of compensatory tachycardia upon diprivanum administration is accounted for by a combined effect of vagotonic diprivanum action and its ability to inhibit moderately the automatic function of the sinoatrial node. PMID- 8239020 TI - [The respirator injector as a possible cause of complications in high frequency artificial respiration of the lungs]. AB - Improper design of an injector can cause pulmonary overdistension, fall in venous return, alveolar hypoventilation, insufficient humidification of inhaled gas and oxygen toxicity syndrome during HFJV. Functional characteristics of widely used injectors were determined and their role in the development of serious complications of HFJV is discussed. Special attention is paid to the flow resistance of the injector, its inner volume, ejection ratio, and maximum inspiratory pressure generated by the injector. Practical steps to avoid these complications are recommended. PMID- 8239021 TI - [Comparative aspects of the use of dalargin in comprehensive anesthesiologic protection during the surgical correction of congenital heart disease]. AB - The use of graded infusion of dalargin (a home-made synthetic analogue of leu enkephalin) in a complex of anesthesiological protection has been analysed in patients with radical correction of tetralogy of Fallot and atrial septal defect. It has been shown that delargin has a pulmonoprotective effect, moreover, it is supposed to possess a certain analgetic activity. Dalargin reduces total pulmonary resistance, the normalizing effect of the drug on the vascular tone and the degree of pulmonoprotecive action being higher in cases when the preceding pathological shifts were more pronounced. The author believes it is indicated to apply the drug in a complex of anesthesiological protection during radical correction of tetralogy of Fallot and atrial septal defect primarily to ensure intraoperative pulmonoprotection and optimization of the vascular tone in the pulmonary artery. PMID- 8239022 TI - [The feasibility of predicting cardiorespiratory complications in thoracic cancer surgery]. AB - Complex functional preoperative investigation performed in 140 patients with lung or esophagus cancer made it possible to identify 2 functional parameters- exercise tolerance and systolic pressure in the pulmonary artery, which when compared with the course of the intra- and postoperative periods, proved most informative for the prediction of respiratory and cardiovascular complications. PMID- 8239023 TI - [Optimization of external respiration in patients during surgery with different types of anesthesia using transcutaneous electric stimulation of the diaphragm]. AB - Inadequate function of respiratory muscles, of the diaphragm among other ones, is responsible, together with other causes, for acute respiratory insufficiency. Transcutaneous electric stimulation of the diaphragm by ESD-2P and ESD-2N-NC devices was used under conditions of intravenous, epidural lidocaine anesthesia, epidural lidocaine+phentanyl, and combined anesthesia in 82 patients during surgery and in 49 patients in the immediate postoperative period. Transcutaneous electric stimulation of the diaphragm essentially improved ventilation volume and regularity, as well as pulmonary gas exchange, particularly so in marked disorders occurring during surgery under intravenous anesthesia. PMID- 8239024 TI - [Electroanesthesia during conservation of spontaneous respiration with an apparatus of novel construction]. PMID- 8239025 TI - [Caudal epidural anesthesia during interventions on the lower extremities in patients with hemophilia]. AB - Anesthesia techniques during surgery on the lower extremities have been compared in 57 patients with hemophilia. It has been shown that caudal epidural anesthesia according to an elaborated technique could be a method of choice. PMID- 8239026 TI - [The status of central and peripheral hemodynamics at different stages of combined peridural anesthesia with lidocaine and promedol in children]. AB - The results of studies of central and peripheral hemodynamics during surgery performed under combined epidural anesthesia with lidocaine and promedol are presented. The study was performed on 82 children with surgical pathology aged 3 to 14 years. The state of hemodynamics was assessed using integral body plethysmography suggested by M. I. Tishchenko. The results of the studies have shown that surgery performed under epidural anesthesia with lidocaine and promedol was associated with stabilization of basic central and peripheral hemodynamic parameters at baseline level, which indicates that there is an effective anesthesiological protection of a child's body from the operation trauma. PMID- 8239027 TI - [The concentration of end products of purine metabolism as possible signs of tissue hypoxia in critically ill surgical patients]. AB - The study was performed on 26 critically ill patients with hemorrhagic pancreonecrosis and disseminated suppurative peritonitis. Central hemodynamic and oxygen transport parameters have been determined in all the patients. To assess the degree of tissue hypoxia end products of ATP catabolism (hypoxanthine, xanthine, and uric acid) have been studied. All the studies were performed at various stages of correction of volemic disturbances and oxygen transport function. It has been established that an increased blood plasma level of hypoxanthine and xanthine may serve as an additional criterion of tissue hypoxia in critically ill surgical patients. Changes in hypoxanthine, xanthine, and uric acid concentrations and oxygen transport parameters can be used to assess changes in the functioning of the microcirculatory bed. PMID- 8239028 TI - [The effect of intravascular helium-neon laser blood irradiation on the state of the compensatory processes in the acute period of hemorrhagic shock and after resuscitation]. AB - The effect of low-intensity intravascular laser blood irradiation on morphofunctional characteristics of erythrocytes and circulation parameters has been studied experimentally on 22 adult inbred anesthetized dogs of both sexes 9 to 23 kg of weight during a 2-hour hemorrhagic shock and in the first hours after resuscitation. It has been established that the use of intravascular laser blood irradiation during 45 min of hemorrhagic shock stabilizes erythrocyte membranes and improves myocardial function. Hyperdynamic and hyperperfusion syndromes were significantly more pronounced in the postresuscitation period in animals treated by intravascular laser blood irradiation than in the control. Thus, the efficacy of intravascular laser blood irradiation was associated both with the recovery of erythrocyte cytoarchitectonics and the enhancement of myocardial contractility. PMID- 8239029 TI - [Clinical experience in the use of dobutamine in patients in the early postoperative period]. PMID- 8239030 TI - [Changes in the content of microelements--copper, zinc and iron--in the blood of patients following cardiopulmonary bypass]. AB - Changes in the blood content of trace elements (E)--copper, zinc and iron--have been studied in 10 patients with ischemic heart disease subjected to cardiopulmonary bypass surgery. It has been shown that blood TE concentrations decreased considerably during surgery. Variability in blood TE levels could be accounted for by hemodilution caused by the administration of colloids and crystalloids that contained no TE under study. These solutions were used initially for filling assisted circulation device. However, it is noteworthy that the degree of hemodilution was not the only reason for the changes in blood TE content, as despite hematocrit normalization it was only blood iron content that returned to preperfusion level. It has been demonstrated that copper and zinc blood concentrations were considerably lower in patients developing heart failure in the early postperfusion period than in those with an uncomplicated postoperative period. It has been shown that the nature and degree of changes in copper, zinc and iron levels in the coronary sinus blood during heart surgery can be assessed by changes in their arterial blood content. PMID- 8239031 TI - [The effect of adenosine on the contractile function of the heart during experimental ischemia and reperfusion]. AB - It has been shown that 30 min ischemia followed by 30 min reperfusion inhibits cardiac contractility. The addition of adenosine at a concentration of 50 or 100 mumol/l into the perfusate normalized a decreased cardiac contractility and prevented the onset of contracture. PMID- 8239032 TI - [A case of unsuccessful repeat epidural anesthesia during surgery on lower limb vessels]. PMID- 8239033 TI - [Evaluation of the efficacy of the work of a pediatric department of resuscitation and intensive care]. PMID- 8239034 TI - [Hemosorption in the combination therapy of acute aconite poisoning]. PMID- 8239035 TI - [Acute renal failure following halothane anesthesia]. AB - Three cases have been reviewed when acute renal failure without signs of concomitant hepatic failure developed post-operatively in patients with a so far intact renal function. A potential pathogenetic role of halothane in the onset of acute renal failure in these patients is discussed. PMID- 8239036 TI - [Resuscitation as a neurologic science]. PMID- 8239037 TI - [A concept of antinociceptive anesthesia]. PMID- 8239038 TI - [Clinical stages and subjective sensations in xenon anesthesia]. AB - Clinical pattern of monoanesthesia with Xe:O2 (70:30) has been studied in 12 anesthesiologists volunteers in comparison with their subjective sensations. Four clinical stages of anesthesia have been identified: paresthesia and hypoalgesia, euphoria and psychomotor activity, analgesia and partial amnesia, anesthesia (analgesia and amnesia). Induction anesthesia took 5 to 6 min, anesthesia discontinuation was easy and prompt without any adverse events. In healthy men it was impossible to achieve complete amnesia with Xe:O2 (60:40). Subjective sensations from the anesthesia were pleasant. Xenone is the best alternative to N2O2. PMID- 8239039 TI - [The surgical treatment of an aneurysm of the celiac trunk and hepatic artery]. AB - We report a case of splenic aneurysms with double location, at the celiac trunks and at the common hepatic artery, in the same patient that was treated by a surgical procedure. We also review literature about the hepatic aneurysms at the celiac trunks, with special attention on the aspects of etiology, natural evolutions and different surgical procedures available. It is remarkable on this kind of pathology that special attention should be paid in order to avoid the simple ligature-exclusion of the aneurysm, it involves realizing an endoaneurysmorrhaphy followed by an aorto-hepatic by-pass. PMID- 8239040 TI - [A recurrent desmoid tumor of the popliteal fossa with vascular compromise]. AB - We report a case of a 18-year-old male patient with his fourth relapse of a desmoid tumour of the left popliteal hollow after a traumatism in this area. Arterial displacement and extrinsic compression were showed by arteriography. Tumoration was extirpated including popliteal artery, in this way a terminal by pass from distal femoral to peroneo-tibial trunks with contralateral great saphena was made. Diagnosis of intestinal polyposis and of the little osseous malformations, was discarded these pathologies frequently are related with these tumours. The research for 8 years and 4 months doesn't show any relapse, nor local neither multicentral. Furthermore, some considerations about this theme are reported. PMID- 8239041 TI - [Pseudomembranous colitis after surgery for a ruptured abdominal aortic aneurysm]. AB - We present a rare postoperative complication after surgical procedures for rupture of abdominal aortic aneurysms. The disease, a pseudomembranous colitis, was early recognized (by evidence of clostridium difficile after a coprocultive) and satisfactorily treated with vancomycin. From the literature review we found only a similar case but results were absolutely different from our case. PMID- 8239042 TI - [The fibrinolytic treatment with urokinase of acute arterial thrombosis]. AB - We present our experiences on 86 patients with acute arterial thrombosis of the legs, undergoing a fibrinolytic treatment with urokinase. Results from the treatment are analyzed according to: the administration way (systemic, locoregional, intrathrombotic), the level of thrombosis (upper or lower legs), the associated morbidity and mortality. PMID- 8239043 TI - [The hemodynamic characterization of the diabetic patient with arterial calcifications]. AB - This study was designed to describe the presence of calcifications according to the clinical features of the diabetic patient and the hemodynamics of the calcified arteries. With this purpose, 197 lower limbs from diabetic patients (type I and II) and carbon-hydrate intolerant patients, were studied. In all of the patients, the pressure ratio leg/arm was measured. On the same way, the arterial flow velocity was recorded using the Doppler ultrasonography on the pedia and postero-tibial arteries. The arterial calcifications, evident on the radiography of the foot, were more frequent between the type I patients and the neuro-infections diabetic foot. According to the hemodynamics point of view, we found a trend of association of more pathologic arterial flow velocity curves with the presence of calcifications (specially on the intima layer). It was also remarkable that an arterial incomprensibility was always associated with arterial calcifications. PMID- 8239044 TI - [Ozone therapy in the advanced stages of arteriosclerosis obliterans]. AB - Fifteen patients with atherosclerosis obliterans at the lower limbs, no candidates to revascularizing surgery were submitted to ozone therapy. An improvement statistically significant was noticed in the treatment groups since amputation ratio decreased (26.7%) and the need of pain's surgery procedures (13.3%) in comparison with the control group (46.7 and 26.7% respectively). Ozone therapy is considered as a good way in the management of the atherosclerosis with obliteration in late period. PMID- 8239045 TI - [A mathematical analysis of the flow-velocity curves in the femoral arteries]. AB - In order to improve the early diagnosis of the aortoiliac injuries, 98 arteries from several supposedly health patients (different ages) and 41 femoral arteries from patients with occlusion at this level (demonstrated by arteriography) were studied. The analysis from the Fourier's series showed highly significant differences between both groups, and so did the comparison of some indexes automatically measured by the Vasoscan VL equip. By multivariant statistics methods was selected the main group of parameters that allows the differentiation between the ill patients and the healthy ones. This procedure can be useful for the physiopathological study and it may be used as a non-invasive method of diagnosis. PMID- 8239046 TI - Aerosol penetration and leakage characteristics of masks used in the health care industry. AB - BACKGROUND: Historically, surgical masks have been worn to protect patients from being infected by large, pathogen-containing aerosol droplets emitted by health care personnel. Today, emphasis has shifted from solely protecting the patient to protecting the health care worker as well. As a result of new procedures used in operating rooms and clinical areas, aerosolized hazardous agents in the submicrometer size range are being produced, posing a potential threat to health care workers. METHODS: Eight surgical masks were tested for aerosol particle penetration through their filter media and through induced face-seal leaks. RESULTS: The percentage of filter penetration ranged from 20% to nearly 100% for submicrometer-sized particles. In comparison, a dust-mist-fume respirator used in industrial settings had significantly less penetration through its filter medium. When the surgical masks had artificially induced face-seal leaks, the concentration of submicrometer-sized particles inside the mask increased slightly; in contrast, the more protective dust-mist-fume respirator showed a fourfold increase in aerosol penetration into the mask with an artificial leak 4 mm in diameter. CONCLUSION: We conclude that the protection provided by surgical masks may be insufficient in environments containing potentially hazardous submicrometer-sized aerosols. PMID- 8239047 TI - The infectious risks of transfusions in the United States: a decision-analytic approach. AB - INTRODUCTION: The development of AIDS as a result of HIV transmission and of cirrhosis as a result of chronic non-A, non-B hepatitis are the greatest infectious risks associated with transfusion in the United States. The goal of this study is to provide explicit quantitative guidelines to determine when the risk of death from all causes associated with transfusion exceeds the risk associated with anemia. METHODS: This study uses a decision-analytic model. RESULTS: On the basis of reported transfusion complication rates and an independent worst-case calculation of the risk of AIDS and non-A, non-B hepatitis, transfusion with red blood cells should be recommended as long as each unit received reduces the patient's risk of dying from anemia by at least 1/1100. DISCUSSION: Because of the relative safety of the blood supply as a result of universal screening and donor deferral, the overestimation in practice of fatal infectious complications, and the possible underestimation of the risk of anemia, undertransfusion has the potential to be as serious a problem in the United States as is overtransfusion. Although caution should be exercised not to undertransfuse when a patient has an appreciable risk of anemia-associated death, we must be wary not to overtransfuse for temporary relief of morbidity. PMID- 8239048 TI - An epidemiologic study of nosocomial infections in a pediatric long-term care facility. AB - OBJECTIVE: To determine the incidence of hospital-acquired (nosocomial) infection in pediatric long-term care facilities. DESIGN: Prospective cohort. SETTING: An 87-bed pediatric long-term care facility. PATIENTS: All patients receiving long term care at Bloorview Children's Hospital during the study period. RESULTS: Infection developed in 40.1% of patients (n = 456). The nosocomial infection rate per 1000 patient days (mean, 7.84) varied substantially, from 1.66 in May 1988 to 16.37 day in April, 1989. The proportional frequencies of infections were as follows: respiratory, 41.6% (37.0% upper, 4.6% lower); urinary tract, 31.0%; skin, 15.6% (gastric tube site 5.0%, other 10.6%), eyes, 6.4%; gastrointestinal, 3.5%; and other, 1.5%. Of those infections for which an organism was recovered (48.5%), pathogens included Escherichia coli (22.5%), Enterococcus (14.8%), Staphylococcus (14.8%), Streptococcus (11.2%), Klebsiella (10.5%), Pseudomonas (10.1%), Proteus (4.3%), yeast (4.3%), Salmonella (0.7%), Clostridium difficile (0.4%), and other (6.2%). CONCLUSIONS: The incidence and nature of infections in pediatric long-term care facilities differs from those in acute care facilities. Physicians should become familiar with the infection rates in the populations whom they treat. Control requires compliance with currently recognized effective strategies as well as innovative practical approaches to respiratory disease. Behavioral problems related to frequent clean, intermittent catheterization in young adults need to be addressed. PMID- 8239049 TI - Natural history of bloodstream infections in a burn patient population: the importance of candidemia. AB - Because of a perceived increase in Candida bloodstream infections in our burn unit, we retrospectively reviewed all the microbiologic data and the medical records of 209 patients with burns admitted during a 42-month period. Twice weekly burn wound cultures demonstrated that Candida species were the tenth most frequently isolated organisms (69/191 patients, 36%). Despite the low frequency of isolation from burn wounds, Candida species were the most common organisms found in blood cultures and urine cultures. Of 49 patients with positive blood cultures, 16 (33%) had clinically significant culture growth of yeasts: Candida albicans, 12; Candida parapsilosis, 2; Candida tropicalis, 1; and Torulopsis glabrata, 1. Patients with candidemia were more likely than patients with blood culture growth of other organisms to have burn wound cultures that grew Candida (15/16 vs 21/33, p = 0.02), larger burns (61% vs 38%, p < 0.001), and death (63% vs 27%, p = 0.02). Multivariate analysis demonstrated that the total number of blood cultures with microorganism growth and large burn size were the greatest independent risk factors for candidemia. These data demonstrate that yeasts are pathogens of major importance in patients with burns, suggesting that in patients with burns who have suspected sepsis and large burn injury or a previous bacteremia, strong consideration should be given to administration of amphotericin B initiation of empiric antibacterial therapy. PMID- 8239050 TI - Postoperative wound infection after total abdominal hysterectomy: a controlled study of the increased duration of hospital stay and trends in postoperative wound infection. AB - BACKGROUND: Total abdominal hysterectomy, a common operative procedure, is infrequently accompanied by incisional wound infection. No recent study has examined the excess hospital stay attributable to such infections. METHODS: This historical cohort study of cases and matched controls was performed in a tertiary care university hospital. RESULTS: During the 5-year study period (1985 to 1989), the infection rate was 10.5 per 100 procedures; patients with infection remained hospitalized 3.55 days longer than did matched control patients (p = 0.0025). CONCLUSION: In this era after the introduction of the diagnosis-related groups for reimbursement, incisional wound infection after total abdominal hysterectomy leads to a significant period of extra hospital stay. PMID- 8239051 TI - Efficacy evaluation of a 4% chlorhexidine gluconate as a full-body shower wash. AB - A 4% chlorhexidine gluconate (Xttrium Laboratories, Chicago, Ill.) was used in a shower bath application to evaluate its merits in reducing resident skin microorganisms. Five volunteers completed a 14-day microbial stabilization period, a 7-day baseline period, and a 5-day test period. Subjects followed a standard protocol, performed five shower washes and were sampled at both the abdominal and inguinal regions immediately after the shower wash as well as at 3 and 6 hours later on days 1, 2, and 5. Chlorhexidine gluconate provided significant microbial reductions from baseline, with greater reductions noted as the study progressed. Use of chlorhexidine gluconate in a shower wash application in conjunction with a preoperative skin-preparation procedure may reduce the probability of surgically associated infections. PMID- 8239052 TI - Description and reporting of statistical methods. AB - A review of the last three volumes of AJIC (1990, 1991, and 1992, numbers 1 through 3) revealed a modest use of statistics in infection control research. Discussion of the statistics most commonly used results in the following recommendations: (1) Researchers who use the t test should report the t test statistic, exact significance probabilities, and a discussion of assumptions and requirements underlying the analysis. (2) A 95% confidence interval for estimated quantities is preferred to p values. (3) The potential impact of nonresponse bias should be included in reports of survey data. (4) An adequate description of statistical methods should be included in all reports of survey data. (5) An adequate description of statistical methods should be included in all research articles. PMID- 8239053 TI - Are some types of needles more likely to transmit HIV to health care workers? PMID- 8239054 TI - Air-fluidized beds and negative-pressure isolation rooms. PMID- 8239055 TI - Diagnosis of high-grade stenosis of innominate artery. AB - Although pathology of the innominate artery (IA) is not frequent, it causes relevant impairment of extracranial circulation and sometimes cerebrovascular events. Definite differentiation of high-grade stenosis from occlusion of the IA has not been obtained until now with continuous-wave (C.W.) Doppler and duplex system ultrasound, and thus the direct evaluation of IA is not reliable by noninvasive methods. In this study the authors suggest an indirect method of evaluation of IA pathology based on the study of carotid, subclavian, and vertebral arteries (VA) with echo-Doppler-color-flow (EDCF) (with linear--phase array probes of 7.5 and 5 MHz). In the last two years they studied 6 patients with IA pathology (2 with occlusion (occ), 1 with stenosis of 80% and 3 with subocclusive stenosis > 90%). In both patients with occ, right VA flow was inverted, and also the right common carotid (CC) was involved (occ in 1 case and inverted flow in the other); in the 3 subocclusive stenoses a lowered systolic flow in the CC was recorded (48 vs 85 cm/sec, 41 vs 77, and 23 vs 109). In the 80% stenosis, besides the inverted flow in VA, only a reduced diastolic flow in CC was also recorded (19 vs 33 cm/sec measured in the left side). All patients with high grade stenosis were successfully confirmed by an angiographic study, including the different degree of stenosis. Five of the 6 patients underwent surgical correction with a restored flow in the previously involved artery. Only 1 patient with occlusion is waiting for surgical correction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239056 TI - Vertebrobasilar insufficiency: correlation of clinical and radiologic findings. AB - Radiological and clinical findings of 20 cases with angiographically proven stenosis or occlusion of the vertebrobasilar system and with clinical signs and symptoms attributable to posterior circulation disorder were studied. A simplified classification of vertebrobasilar arterial disease was presented; type I refers to stenosis of the vertebrobasilar system or subclavian artery, type II refers to vertebrobasilar occlusion, and type III refers to vertebrobasilar branch disease. Type I lesions were further subdivided into types Ia and Ib; type Ia cases showed no infarction in the posterior territory, whereas type Ib cases showed one or more infarctions on computed tomography or magnetic resonance imaging. This angiographic classification is based on treatment options that can be adopted to specific types of lesion. In type I cases, lesions are usually amenable to surgical or angiographic intervention. In type II cases, short-term anticoagulation is the treatment of choice. In type III cases, treatment is primarily aimed at prevention of complication and further stroke. PMID- 8239057 TI - A botanical compound, Padma 28, increases walking distance in stable intermittent claudication. AB - Thirty-six patients with a median age of sixty-seven years and a median duration of intermittent claudication of five years were randomized to either active treatment with Padma 28 or placebo. The effect of treatment was quantified by measurements of systemic and peripheral systolic blood pressures and by measurements of the pain-free and the maximal walking distance on a treadmill. The ankle pressure index (ankle systolic pressure/arm systolic pressure) was calculated. The group randomized to active treatment received two tablets bid containing 340 mg of a dried herbal mixture composed according to an ancient lamaistic preparation (Padma 28). After active treatments, administered over a period of four months in a double-blinded, randomized design, the patients allocated to this group attained a significant increase in the pain-free walking distance from 52 m (20-106) to 86 m (24-164; P < 0.05) and in the maximal walking distance from 115 m (72-218) to 227 m (73- > 1,000; P < 0.05). The patient-group receiving placebo treatments did not show any significant changes in either the painfree or the maximal walking distance. The authors could not demonstrate any significant changes in the ankle pressure index either during active or during placebo treatment. In conclusion, this study has shown that treatment with Padma 28 over a period of four months significantly increased the walking distance in patients with stable, intermittent claudication of long duration. PMID- 8239058 TI - Endomyocardial disease and eosinophilia. AB - The authors discuss the hypereosinophilic syndrome:incidence, terminology, cytotoxicity, clinical presentation and diagnosis, prognosis, and treatment, and they provide a literature review. PMID- 8239059 TI - Myocardial hypertrophy, angiotensin, and ACE inhibitors. AB - The renin-angiotensin system has long been known as a potent determinant of cardiovascular homeostasis and a powerful regulator of vascular hemodynamics. Over the last twenty years, it has become clear that components of the renin angiotensin system are present in and, in many instances, synthesized in local tissues. The role of some of these local renin systems is now becoming clear, but the role, if any, of local production of angiotensin II in the heart and vasculature remains unknown. Recent evidence indicates that angiotensin II can serve as a growth factor for a variety of cell types including those in the cardiovascular system, and it thus appears possible that systemically or locally produced angiotensin II could subserve important functions in the determination of heart and vasculature structure. Clinical studies tend to corroborate this possibility. The potent effects of converting enzyme inhibition on the regression of left ventricular hypertrophy, the demonstration that the administration of these agents following myocardial infarction reduces detrimental cardiac remodeling, and the finding that converting enzyme inhibition prolongs survival in congestive heart failure patients all point to an important role of angiotensin II on cardiovascular function and raise the distinct possibility that angiotensin II growth effects are involved in these beneficial responses. Additionally, recent data demonstrate conclusively that under certain circumstances and in certain cell types locally produced angiotensin II can serve as an autocrine growth regulatory factor, further adding support to the idea that local renin systems may play an important role in the determination of cardiovascular structure. PMID- 8239060 TI - Changes of soft-tissue water examined with magnetic resonance and electrical impedance tomography: an in vivo experiment. AB - The purpose of this study was the examination of short-term changes in soft tissue water under in vivo conditions. In 12 healthy volunteers the water distribution of a selected slice of the lower limb was examined after change of position and rapid infusion of 0.9% saline solution. Two methods were used for data acquisition: magnetic resonance imaging (MRI) (Repetition Time (TR) = 1600 msec, Inversion Time (TI) = 300 msec, Echo Time (TE) = 22 msec) and electrical impedance tomography (EIT). In orthostatic equilibrium all 12 volunteers demonstrated changes of MR signal intensity and resistivity in the range of 12% and 7%, respectively. After infusion, mean changes of signal intensity and resistivity were 7% and 4%, respectively. The overall correlation of MRI and EIT was -0.96 (p < 0.001). Results indicate that inversion recovery sequences are a sensitive method of detecting in vivo changes of water distribution in selected areas of the body. PMID- 8239061 TI - Vasodilator effects of hydroxylamine in the isolated rodent lung. AB - Hydroxylamine is a natural product of cellular metabolism that possesses vasodilating properties similar to those of endothelium-derived relaxing factor (EDRF). In the rodent pulmonary circulation preconstricted with the endoperoxide analog U-46619, hydroxylamine relaxed the vasculature in a concentration dependent manner. Blockade of the hydroxylamine vasodilator response by methylene blue indicated that the mechanism of vasorelaxation is similar to that of EDRF. In this preparation, hydroxylamine is a more potent vasodilator than nitroglycerin. PMID- 8239062 TI - Atrial natriuretic peptide and left atrial systolic function in normal subjects. AB - The purpose of this study is to elucidate factors determining the release of atrial natriuretic peptide (ANP) in normal volunteers at rest. Ten normal volunteers were included in this study. Ages ranged between twenty-seven and thirty-three years (mean twenty-seven). The authors measured plasma levels of ANP, and four cardiac chamber volumes, and their functions by cine magnetic resonance imaging using the rephased gradient-echo method. Plasma levels of ANP in 10 normal subjects were 15.0-37.1 pg/mL (mean 23.6 pg/mL). No significant relationship was seen between ANP level and heart rate or blood pressure. The level of ANP was positively correlated with left atrial (LA) emptying fraction and negatively correlated with LA minimal volume (r = 0.85, P < 0.01; r = 0.64, P < 0.05, respectively). No significant relationship was observed between plasma ANP level and left ventricular, right ventricular, or right atrial parameters. These results suggest that LA systolic function is one of the major determinants for ANP release in normal subjects at rest. PMID- 8239063 TI - Diffuse arterial aneurysms in a case of Ehlers-Danlos syndrome--a case report. AB - A three-year-old boy with the diagnosis of Ehlers-Danlos syndrome (EDS) with persistent ductus arteriosus and multiple diffuse arterial aneurysms is presented. The case is classified as "EDS type unknown" because the clinical features and the inheritance pattern differ from the types described previously. It is stressed that the diagnosis of the disease is important for genetic counseling and surgical intervention. PMID- 8239064 TI - Right atrial thrombus mimicking myxoma and bilateral pulmonary artery aneurysms in a patient with Behcet's disease--a case report. AB - A case of Behcet's disease, in which superior vena cava syndrome was the presenting feature is reported. Magnetic resonance imaging and echocardiography revealed a mass lesion in the right atrium. This patient developed bilateral pulmonary artery aneurysms postoperatively and represents one of the bizarre manifestations of Behcet's disease. PMID- 8239065 TI - Clinically isolated left coronary ostial stenosis: a manifestation of cardiovascular syphilis--case history. AB - The authors report a case of clinically isolated left coronary ostial stenosis in a thirty-seven-year-old man as a manifestation of cardiovascular syphilis. Notably he was free of the usual risk factors for coronary artery disease, and the rest of the coronary tree was angiographically normal. PMID- 8239066 TI - Recurrence of a left ventricular thrombus after surgical excision in a young patient with normal left ventricular systolic function--a case report. AB - The case report of a young patient with recurrence of intraventricular thrombus represents an unusual presentation with no apparent clinical explanation despite echocardiographic, surgical, and pathologic confirmation. The other unusual feature of this case is the absence of predisposing factors for intraventricular thrombus. The patient developed the second left ventricular thrombus after he stopped taking anticoagulants. The authors' recommendation for such a patient is that he continue to receive anticoagulants for life. PMID- 8239067 TI - Isolation and characterization of cDNA clones for chicken major histocompatibility complex class II molecules. AB - The chicken major histocompatibility complex (MHC), the B complex, is being intensively analysed at the DNA level. To further probe the molecular structure of chicken MHC class II genes, cDNA clones coding for chicken MHC class II (B-L) beta chain molecules were isolated from an inbred G-B2 Leghorn chicken spleen and liver. Twenty-nine cDNA clones were isolated from the spleen and eight cDNA clones were isolated from the liver. Based on restriction maps, most clones could be clustered into one family of genes. Four cDNA clones were sequenced (S7, S10 and S19 from the spleen and L1, which was identical to S19, from the liver). Complete amino acid sequences of B-L beta chain molecules were predicted from the nucleotide sequences of the cDNA clones. Although both the nature and the location of the conserved residues were similar in chicken and mammalian sequences, some species-specific differences were found, suggesting that the structures of the B-L molecules of this haplotype are similar, but not identical, to their mammalian counterparts. PMID- 8239068 TI - DNA profiling of cattle using micro- and minisatellite core probes. AB - We have evaluated 15 different micro- and minisatellite core probes for use in identity and paternity testing in cattle, based on Southern blot hybridization analysis. The core probes were tested in animals of different breeds and by analysis of seven two-generation pedigrees. Of the 15 core probes tested, seven were able to detect on average seven variant bands per individual animal. Segregation analysis showed that on average two out of 36 variant bands scored per core probe were genetically linked while two out of 12 variant bands correspond to the same allelic pair. The results obtained demonstrate the effectiveness of multilocus core probes for determining identity and paternity in cattle. PMID- 8239069 TI - Synteny mapping in the horse using horse-mouse heterohybridomas. AB - In a study of 35 horse-mouse heterohybridoma cell lines, synteny in the horse was found between LDHB, PEPB and IGF1 and between NP, MPI and IDH2. A synteny between ADA and PEPC was also indicated. The loci for horse immunoglobulin light chain (IgL) genes and for LDHA were independent. PMID- 8239070 TI - Preparation of chromosome-specific paints and complete assignment of chromosomes in the pig flow karyotype. AB - Sorted chromosomes from each of the 20 clusters of the male porcine bivariate flow karyotype were amplified and biotinylated using DOP-PCR. The chromosomes comprising each cluster were identified by fluorescence in situ hybridization (FISH) of the 20 probes to R-banded male pig metaphase spreads. A standard flow karyotype for the pig is presented. PMID- 8239071 TI - Strong association between polymorphisms in an intronic microsatellite and in the coding sequence of the BoLA-DRB3 gene: implications for microsatellite stability and PCR-based DRB3 typing. AB - A highly polymorphic microsatellite in the bovine DRB3 gene was characterized by polymerase chain reaction (PCR) analysis and DNA sequencing. A very strong association between expressed DRB3 polymorphism and microsatellite alleles was revealed by PCR analysis of genomic DNA from 116 animals representing three breeds of cattle. The results indicated a low frequency of microsatellite length mutations as the association was consistent over breeds. The DRB3 microsatellite may be utilized in a PCR-based typing method of bovine class II alleles. The microsatellite polymorphism did not distinguish all known DRB3 alleles, but it was shown that this method may be complemented by the use of allele-specific PCR based on the extensive polymorphism in the DRB3 exon 2. The DNA sequences of seven microsatellite alleles, associated with different class II haplotypes, were determined. The DRB3 microsatellite is composed of three repeat motifs, a stretch of at least 10 uninterrupted (TG)n dinucleotides, a long but interrupted stretch of (GA)n dinucleotides, and a few (CAGA)n tetranucleotides. There were pronounced sequence differences between alleles and the results indicated that the evolution of this microsatellite has involved length mutations of the dinucleotide repeats as well as point mutations causing interruptions in the dinucleotide repeats. PMID- 8239072 TI - Isolation and characterization of microsatellites from the canine genome. AB - Microsatellite sequences comprising (dC-dA)n.(dG-dT)n repeats have been isolated from canine libraries and sequenced. Oligonucleotide primers have been synthesized to the microsatellite flanking sequences and used in the polymerase chain reaction to amplify those loci from genomic DNA. The degree of polymorphism of each microsatellite was estimated in a set of unrelated dogs. It is concluded that of the 10 loci studied, nine are sufficiently polymorphic to be useful in genetic studies. PMID- 8239073 TI - The sex-determining region Y (SRY) gene is mapped to p12-p13 of the Y chromosome in pig (Sus scrofa domestica) by in situ hybridization. AB - The sex-determining region Y is a gene located in the distal portion of the short arm of human (SRY) and mouse (Sry) Y chromosomes and considered to be the best candidate for the testis determining factor (TDF/Tdy). The gene is believed to be the key factor in sex differentiation in mammals and is conserved across mammalian species. We report herein that the SRY/Sry gene has been assigned to p12-p13 on the short arm of the Y chromosome in pig by in situ hybridization. The result confirms interspecies conservation of this chromosomal segment in the evolution of mammalian chromosomes, and suggests further use of this gene probe in genomic studies in another mammals. The assignment of the Sry gene is the second physical gene mapping data available for the Y chromosome in pigs. Such data can be used in the effort of constructing the pig gene map and for further establishment of a comparison of sex chromosome morphology in different mammalian species concerning sex-specific and pseudoautosomal regions. PMID- 8239074 TI - Linkage between the loci for serum albumin and vitamin D binding protein (GC) in sheep. AB - Evidence for close genetic linkage between the loci for serum albumin (ALB) and vitamin D binding protein (GC) in sheep is presented. No recombinants were found in 28 informative offspring of a single ram family. The recombination frequency between the two loci was estimated to be in the range of 0 to 10%. No sign of linkage was observed between the ALB-GC complex and transferrin. PMID- 8239075 TI - A new, partially deficient transferrin variant in the pig. AB - A new, partially deficient transferrin variant (TFF) was found in serum samples of a wild boar and his offspring from crossing with Pietrain sows. Family analyses confirmed its genetic control by a codominant allele, TFF. Finding of this new variant has brought the total number of pig transferrin variants to nine (F, I, A, B, C, X, D', E, D). PMID- 8239076 TI - Localization of the tumour protein, TP53, on porcine chromosome 12q12-q14. AB - A human cDNA probe of the tumour protein p53 (TP53) was used to localize the homologous porcine gene by in situ hybridization. The gene was mapped to chromosome 12q12-q14. Together with already known mapping data, these results confirm the localization of an evolutionary conserved linkage group on porcine chromosome 12 which is localized in man on chromosome 17, in cattle on chromosome 19, and in mice on chromosome 11. PMID- 8239077 TI - A fast and highly sensitive method for detecting freemartinism in bovine twins using immunomagnetic beads and Y-specific PCR primers. AB - A method for detecting male cells in the blood of the female calf in bovine heterosexual twin pregnancies has been established. Nucleated cells were isolated from full blood by immunomagnetic separation, lysed by boiling and then subjected to polymerase chain reaction (PCR) amplification with Y chromosome specific primers. Diagnosis was achieved within one day. The method was successfully used on blood samples that had been stored at +4 degrees C for more than one month. Dilution of male blood in female blood showed that XY cells were detectable down to a concentration of 0.1%. This method should be amenable to automatization and can be adapted to any PCR-based genetic test. PMID- 8239078 TI - An additional locus (PI4) in the protease inhibitor (PI) gene cluster of pigs. AB - A further alpha-protease inhibitor system, PI4, was detected in porcine sera using either 2D agarose gel, pH 5.0-PAGE, pH 9.0, or 1D PAGE followed by immunoblotting with rabbit anti-porcine PI2 or PI3 antisera. PI4 inhibited chymotrypsin, but not trypsin. Seven allelic variants of PI4 were described. By haplotyping of alpha-protease inhibitor systems in 52 complete families it was shown that PI4 locus belongs to the PI gene cluster. The probable order of the PI loci was: PI1, PO1A, PI2, PI4, PI3. PMID- 8239079 TI - Silent blood chimaerism in a mare confirmed by DNA marker analysis of hair bulbs. AB - Microsatellite DNA markers in a mare's hair bulbs not concordant with markers in her blood confirmed the hypothesis of chimaerism which had been proposed to explain the apparent parentage exclusion of the mare from her suckling foal. Parentage analysis for this foal based on genetic markers not originating from blood cells of its dam supported a parentage verification conclusion. PMID- 8239080 TI - Characterization of the 3' untranslated region of bovine parathyroid hormone gene in N'Dama and Boran cattle. AB - We describe a polymorphism in the bovine gene PTHG which can be readily typed by PCR assay. The polymorphism is codominantly inherited and the allele frequencies appear characteristic of Bo indicus and B. taurus cattle. PMID- 8239081 TI - Dinucleotide repeat polymorphism at the bovine HUJ614 locus. PMID- 8239082 TI - Dinucleotide repeat polymorphism at the bovine HUJ616 locus. PMID- 8239083 TI - Dinucleotide repeat polymorphism at the bovine HUJ625 locus. PMID- 8239084 TI - A dinucleotide repeat polymorphism at the adenylate cyclase activating polypeptide locus in sheep. PMID- 8239085 TI - Canine tetranucleotide repeat polymorphism at the VIAS-D10 locus. PMID- 8239086 TI - Special issue introduction: in honor of Fred S. Grodins. PMID- 8239087 TI - Chemoreceptor drives and short sleep-wake cycles during hypoxia: a simulation study. AB - We used a Grodins-type mathematical model of the cardio-pulmonary system to investigate the cycling behaviour of the respiratory system during hypoxia in response to changes of state between waking and sleeping, namely a diminution of respiratory chemosensitivity during sleep. Shifts between waking and sleeping were triggered by various combinations of threshold values for PAO2. Mild or moderate hypoxia was simulated by values of inspired O2 concentration between 13% and 16% (normal 21%). In mild or moderate hypoxia, reductions of overall respiratory gain from about 2 to 0.8 l.min-1 mmHg-1 at sleep onset will produce falls in PAO2 likely to cause sleep-wake cycles with oscillations in PAO2. The higher the arousal threshold (in relation to steady-state PAO2 during sleep), the shorter and more stable the sleep-wake cycles. As the arousal threshold is raised, and as hypoxia is exacerbated from mild (FIO2 = 16%) to moderate (FIO2 = 13%), the sleep-wake cycle length tends to converge to a value around one minute. The level and determinants of the "back-to-sleep" threshold are hard to define from presently available experimental data, but the level is not important in determining the length of the sleep-wake cycle compared to the arousal threshold. Alinearities in chemoreceptor feedback were introduced first by incorporating "drive" thresholds, to simulate central or obstructive apnoea. This produced larger oscillations in respiratory variables, but no change in cycle length. Chemoreceptor thresholds for PCO2 at the level of 38-39 mmHg did produce shorter ventilation cycles, down to about 20 s in length, but these were not related in any simple way to the resulting sleep-wake cycles. The combination of sleep state changes and chemoreceptor feedback alinearities can produce short sleep-wake cycles despite the diminution in chemoreceptor gain occurring in sleep. PMID- 8239088 TI - Modeling human diaphragmatic electromyogram and airflow responses to imperceptible mechanical loads. AB - We used an esophageal electrode to measure the amplitude and timing responses of diaphragmatic electrical activity and airflow in response to flow resistive and elastic loads at or below the threshold for conscious detection, applied pseudorandomly to the oral airway of eight normal human subjects. The mechanical and neural parameter responses to mechanical loading were cross-correlated with the pseudorandom loading sequence to obtain estimates of the impulse responses. We convolved the resultant impulse response estimates with the loading sequence to obtain the responses predicted from the linear component of the generalized Wiener kernel model. Highly significant correlations and close correspondence were found between the model-predicted and ensemble-averaged experimental responses for nearly all neural and mechanical parameters in all subjects. For nearly every aspect of the pattern, with few exceptions, the response to these small load perturbations in all eight subjects was adequately explained by an impulse response, leaving negligible nonlinearity to require higher-order cross correlations. These results indicate that the estimated impulse responses accurately model the dynamics of the neural and mechanical responses in human subjects for the types and magnitudes of loads applied. This study supports use of the pseudorandom loading technique to determine the neural and mechanical responses to imperceptible mechanical loads in conscious humans. PMID- 8239089 TI - Influence of the viscoelastic properties of the respiratory system on the energetically optimum breathing frequency. AB - We hypothesized that the viscoelastic properties of the respiratory system should have significant implications for the energetically optimal frequency of breathing, in view of the fact that these properties cause marked dependencies of overall system resistance and elastance on frequency. To test our hypothesis we simulated two models of canine and human respiratory system mechanics during sinusoidal breathing and calculated the inspiratory work (WI) and pressure-time integral (PTI) per minute under both resting and exercise conditions. The two models were a two-compartment viscoelastic model and a single-compartment model. Requiring minute alveolar ventilation to be fixed, we found that both models predicted almost identical optimum breathing frequencies. The calculated PTI was very insensitive to increases in breathing frequency above the optimal frequencies, while WI was found to increase slowly with frequency above its optimum. In contrast, both WI and PTI increased sharply as frequency decreased below their respective optima. A sensitivity analysis showed that the model predictions were very insensitive to the elastance and resistance values chosen to characterize tissue viscoelasticity. We conclude that the WI criterion for choosing the frequency of breathing is compatible with observations in nature, whereas the optimal frequency predictions of the PTI are rather too high. Both criteria allow for a fairly wide margin of choice in frequency above the optimum values without incurring excessive additional energy expenditure. Furthermore, contrary to our expectations, the viscoelastic properties of the respiratory system tissues do not pose a noticeable problem to the respiratory controller in terms of energy expenditure. PMID- 8239090 TI - Adaptive neural network that subserves optimal homeostatic control of breathing. AB - An adaptive neural network model that exhibits the optimality and homeostasis characteristics of the respiratory control system is described. Based upon the Hopfield network structure and a postulated Hebb-like respiratory synapse with correlational short-term potentiation, the model is capable of mimicking the normal ventilatory responses to exercise and CO2 inputs without the need for an explicit exercise stimulus. Results suggest the possibility of an adaptive neuronal mechanism that effects optimal homeostatic regulation of respiration in mammals. PMID- 8239091 TI - Perceptual contributions to optimization of breathing. AB - Besides regulating the energetic cost of breathing, optimization of breathing may involve the alleviation of an uncomfortable breathing sensation. In this article, we consider perceptual contributions to the optimization of breathing. Just as the cost function proposed by Poon depends on ventilation and arterial PCO2 so does the sense of dyspnea. Consequently, we examined the relationship between breathing discomfort and Poon's cost function. Based on our model and psychophysical studies, it appears that breathing discomfort, which can integrate chemical and mechanical inputs to the respiratory controller, may not operate simply as an estimate of a cost function. We explain how our reflex control model can dynamically minimize a cost function such as Poon's. Also, we consider the influence of willful adjustments of ventilation on breathing discomfort. From this we infer that ventilatory optimization may emerge from automatic reflexes and behavioral responses that involve excitatory chemical and inhibitory neuromechanical feedbacks. PMID- 8239092 TI - Optimal ventilatory patterns in periodic breathing. AB - The goal of this study was to determine whether periodic breathing (PB), which is highly prevalent during sleep at high altitudes, imposes physiological penalties on the respiratory system in the absence of any accompanying disease. Using a computer model of respiratory gas exchange, we compared the effects of a variety of PB patterns on the chemical and mechanical costs of breathing to those resulting from regular tidal breathing. Although PB produced considerable fluctuation in arterial blood gas tensions, for the same cycle-averaged ventilation, higher arterial oxygen saturation and lower arterial carbon dioxide levels were achieved. This result can be explained by the fact that the combination of large breaths and apnea in PB leads to a substantial reduction in dead space ventilation. At the same time, the savings in mechanical cost achieved by the respiratory muscles during apnea partially offset the increase during the breathing phase. Consequently, the "pressure cost," a criterion based on mean inspiratory pressure, was elevated only slightly, although the average work rate of breathing increased significantly. We found that, at extreme altitudes, PB patterns with clusters of 2 to 4 large breaths that alternate with apnea produce the highest arterial oxygenation levels and lowest pressure costs. The common occurrence of PB patterns with closely similar features has been reported in sleeping healthy sojourners at extreme altitudes. Taken together, these findings suggest that PB favors a reduction in the oxygen demands of the respiratory muscles and therefore may not be as detrimental as it is generally believed to be. PMID- 8239093 TI - Uniqueness of optimal controllers during exercise. AB - The optimization hypothesis of respiratory control during exercise is based on the minimization of a function reflecting both chemical and mechanical costs. Both additive and multiplicative controllers have been derived as optimal from similar cost functions. The purpose of the present study was to explore the uniqueness of such predictions. Various formulations of controllers compatible with isocapnia were found to yield identical costs as controllers predicted to be optimal. It was concluded that controller predictions based on optimization theory are not unique. Optimization can occur with either an additive or multiplicative controller or any combination of the two which satisfies an isocapnic constraint. A general form of a combined additive-multiplicative controller was derived which was found to be compatible with previously reported experimental data collected during combined CO2 inhalation and exercise. PMID- 8239094 TI - Mechanisms of ventilatory periodicities. AB - The study of ventilatory periodicities is relevant to the problem of obstructive sleep apnea. Apneas occur at the nadirs of periodicities during sleep. Periodicities can be caused by chemical instability, related to unstable action of the closed loop feedback system for the chemical regulation of breathing. Such instability occurs when overall loop gain is greater than or equal to unity and the phase lag around the loop is 180 degrees. Periodic breathing during hypoxia and in patients with congestive heart failure is likely to be explained by this mechanism. Periodic breathing can also be the result of state instability. Here ventilation declines at sleep onset and the resultant changes in blood gases trigger an arousal, i.e., sudden transition to a lighter stage of sleep. With arousal, ventilation increases. Thus, periodic breathing is secondary to these changes in sleep state. These processes, chemical instability and state instability, can interact and produce complex patterns of oscillation in ventilation. PMID- 8239095 TI - Regulation of PaCO2 during rest and exercise: a modeling study. AB - A nonlinear mathematical model of the CO2 control system was used to examine a number of issues concerning the regulation of PaCO2 during rest and exercise. To gain insight to the regulatory properties of the respiratory system, the open loop gain (GL) and closed loop sensitivities SI = delta PaCO2/delta PICO2 and SV = delta PaCO2/delta VCO2 were calculated. GL indicates the ability of a control system to regulate the controlled variable, PaCO2 in the model. SI and SV represent the change in PaCO2 to unit changes in PICO2 and VCO2, respectively. Model predications were obtained for rest and various intensities of exercise for the following challenges to the respiratory system: (a) CO2 inhalation, (b) i.v. CO2 loading, (c) application of an external dead space, and (d) a shift in the resting operating point. Increasing exercise intensity produced a substantial decrease in GL and increase in SI consistent with the hypothesis that exercise degrades the ability of the respiratory system to regulate PaCO2. However, SV decreased indicating that the respiratory system would actually be better able to regulate PaCO2 if there were fluctuations in VCO2. Thus, GL does not completely describe the regulatory characteristics of the respiratory control system. It is demonstrated that the regulatory characteristics of the respiratory system as described by GL, SI, and SV are complex and depend on the nature of the challenge. Techniques for systematically describing the regulatory properties of the CO2 control system are described. PMID- 8239096 TI - Effects of vagal tone on resuscitation from experimental electromechanical dissociation. AB - STUDY OBJECTIVE: To determine if complete loss of vagal tone improves the return of spontaneous circulation rate in a canine model of electromechanical dissociation cardiac arrest. DESIGN: Prospective, controlled laboratory investigation using an asphyxial canine cardiac arrest model randomized to receive bilateral cervical vagotomy or no vagotomy. INTERVENTIONS: After the production of asphyxial electromechanical dissociation arrest, 16 mongrel dogs remained in untreated electromechanical dissociation for ten minutes and then were randomized to receive bilateral cervical vagotomy or no vagotomy. All animals received standard external CPR and epinephrine (0.02 mg/kg every five minutes) throughout resuscitation. MEASUREMENTS AND MAIN RESULTS: Return of spontaneous circulation was achieved in 13% (one of eight) of no vagotomy and 75% (six of eight) of vagotomy animals (P = .02). Survival to one hour was achieved in 100% (one of one) of no vagotomy and 83% (five of six) of vagotomy animals (P = NS). The hemodynamic and arterial blood gas values at five, ten, and 15 minutes into resuscitation were not significantly different between groups. CONCLUSION: In this canine model of asphyxial electromechanical dissociation cardiac arrest, surgical vagotomy and complete loss of vagal tone improved the rate of return of spontaneous circulation. PMID- 8239097 TI - Chemical immobilization and killing of intra-aural roaches: an in vitro comparative study. AB - STUDY OBJECTIVE: The treatment of live insects in patients' ears is controversial. To determine which chemical agent is most effective for immobilizing and killing intra-aural cockroaches, we carried out the following investigation. DESIGN: An in vitro blinded comparative study. INTERVENTIONS: A model was developed in which live cockroaches were submerged in microscope immersion oil, 2% lidocaine, 4% lidocaine, or 2% viscous lidocaine in a glass beaker. Responses of cockroaches were recorded with a video-cassette recorder and evaluated later by a blinded observer. Measured variables were time to death and time-integrated activity before death. Analysis was by analysis of variance with Tukey's procedure. RESULTS: Four groups of 40 cockroaches each were exposed to each of the four agents. Microscope oil killed the insects most quickly (mean, 27.2 seconds; 95% confidence interval, 23.8 to 30.6). The other agents required more than 40 seconds and were inferior to oil (F = 15.5, P < .0001). Total activity was also least in the microscope oil group (F = 25.7, P < .0001). CONCLUSION: Microscope immersion oil is the most effective agent for immobilizing and killing intra-aural cockroaches. PMID- 8239098 TI - Tetracaine protects against cocaine lethality in mice. AB - STUDY HYPOTHESIS: Tetracaine will enhance cocaine toxicity. STUDY POPULATION: Two hundred forty female Swiss albino mice weighing 27 to 45 g. METHODS: Intraperitoneal injections of tetracaine and cocaine were given to groups of ten mice each in a controlled, blinded fashion. Either tetracaine or an equal volume of normal saline was given five minutes before one of six incremental doses of cocaine, ranging from 60 to 110 mg/kg. The experiment was repeated twice using two different doses of tetracaine: either an LD10 (40 mg/kg determined from preliminary studies), or one-twentieth of the dose of cocaine (which approximates the ratio used in tetracaine, epinephrine, and cocaine). Lethality was recorded at 24 hours. Lethality between groups was compared with a Wilcoxon sign-rank test. RESULTS: Tetracaine reduced cocaine lethality at all doses. This reduction in lethality was statistically significant at both tetracaine doses (P < .05). CONCLUSION: In the mouse, pretreatment with tetracaine significantly decreases cocaine lethality. PMID- 8239099 TI - Prevention of toxicity from oral cocaine by activated charcoal in mice. AB - STUDY OBJECTIVE: To study the effectiveness of activated charcoal in preventing toxicity after an enterally administered cocaine hydrochloride overdose in mice. DESIGN: A prospective, randomized, controlled animal laboratory investigation. INTERVENTIONS: Fasted mice were given aqueous cocaine hydrochloride (0.8% final concentration) 100 mg/kg body weight orally by gavage tube. One minute later, animals received one of three treatments by gavage: 1 g activated charcoal/kg body weight, 2 g activated charcoal/kg body weight, or an equivolume of water (control). All treatments consisted of 20 mL/kg body weight of an activated charcoal slurry with water. MEASUREMENTS: After 24-hour observation, proportions of seizures and deaths between each group were compared using Pearson chi 2 test followed by Fisher's exact test (P < .017 for significance after Bonferroni's correction). MAIN RESULTS: There were 20 seizures and 16 deaths in the control group (20 mice). There were four seizures (P = .0004) and one death (P = .0004) in the 1-g activated charcoal/kg group (ten mice) and five seizures (P = .0018) and three deaths (P = .015) in the 2-g activated charcoal/kg group (ten mice). CONCLUSION: In this mouse model, activated charcoal decreased the incidence of seizures and death after an enteral cocaine hydrochloride overdose. PMID- 8239100 TI - Prevalence of sorbitol in multiple-dose activated charcoal regimens in emergency departments. AB - STUDY OBJECTIVES: To determine the availability and use of premixed activated charcoal in sorbitol preparations during multiple-dose activated charcoal therapy in the emergency department. DESIGN AND SETTING: A prospective telephone survey of all 911 receiving hospitals within the catchment area of one poison center. TYPE OF PARTICIPANTS: Hospital pharmacy supervisors and ED charge nurses. INTERVENTION: Hospital pharmacy supervisors were surveyed about the available preparations of activated charcoal on their hospital's formulary, and ED charge nurses in these same hospitals were surveyed about the prevalence of sorbitol use in multiple-dose activated charcoal regimens. MEASUREMENTS AND MAIN RESULTS: Eleven hospitals (16%) stocked only activated charcoal in sorbitol preparations. Twenty-one hospitals (31%) had both activated charcoal in sorbitol preparations and activated charcoal without sorbitol preparations, and 35 hospitals (52%) had only activated charcoal without sorbitol preparations. Repeat dosing of sorbitol during multiple-dose activated charcoal therapy occurred in 33 of 67 (49%) of the EDs surveyed. CONCLUSION: Sorbitol dosing is often repeated with activated charcoal during multiple-dose activated charcoal therapy in the ED because of the ready availability (and sometimes exclusive availability) of premixed activated charcoal in sorbitol preparations. PMID- 8239101 TI - A clinical trial of escalating doses of flumazenil for reversal of suspected benzodiazepine overdose in the emergency department. AB - STUDY OBJECTIVE: To determine if flumazenil, when used in doses higher than those currently recommended, could reverse the effects of a benzodiazepine (BDZ) overdose in patients who might not otherwise respond and whether the higher dose was associated with increased adverse effects. DESIGN: Multicenter, randomized, double-blind, placebo-controlled, balanced, with parallel groups. Open-label flumazenil administration was available if a patient failed to respond or became resedated. SETTING: Sixteen emergency departments in the United States. POPULATION: Patients presenting to the ED with clinically significant signs and symptoms of a known or suspected BDZ overdose. INTERVENTIONS: Patients were randomized to receive 10 mL/min of placebo or flumazenil (1 mg/10 mL) each minute for ten minutes. If there was no response, up to 3 mg of open-label flumazenil could be administered. MEASUREMENTS AND MAIN RESULTS: Of 170 patients enrolled, 87 received flumazenil and 83 received placebo. The demographic characteristics of both groups were comparable. Ten minutes after the beginning of study drug infusion, patients were evaluated using the Clinical Global Impression Scale (CGIS), Glasgow Coma Scale (GSC), and Neurobehavioral Assessment Scale (NAS). The mean +/- SD CGIS score at ten minutes for BDZ-positive patients was 1.41 +/- 0.72 for patients who received flumazenil and 3.41 +/- 0.91 for the placebo group (P < .01). There was no difference in the mean CGIS score between the flumazenil (3.25 +/- 1.15) and placebo (3.75 +/- 0.69) groups in BDZ-negative patients. The GCS and NAS were also significantly better in patients who were BDZ-positive and received flumazenil. The mean +/- SD dose of flumazenil administered during the double-blind phase was 71.3 +/- 34.2 mL (7.13 mg) compared with 95.06 +/- 16.03 mL of placebo. Of the 39 patients who had BDZ-positive drug screens and received flumazenil, 29 (74%) responded to 3 mg or less. Six additional patients responded to 4 or 5 mg, and one patient responded to 8 mg. The most common adverse effects in patients who received flumazenil were injection site pain (10.3%), agitation (8%), vomiting (3.4%), dizziness (3.4%), headache (3.4%), tachycardia (3.4%), and crying (3.4%). Three patients developed seizures. Two were associated with significant tricyclic antidepressant overdoses and one with propoxyphene ingestion. Two patients had positive drug screens for BDZ. CONCLUSION: Flumazenil rapidly and effectively reverses the clinical signs and symptoms of a BDZ overdose. Most patients will respond to 3 mg or less, but a small number may require a higher dose for reversal of clinical symptoms. Patients with concomitant tricyclic antidepressant overdose may be at risk for developing seizures. PMID- 8239102 TI - Poisoning deaths not reported to the regional poison control center. AB - STUDY OBJECTIVES: This study was designed to characterize poisoning deaths in Rhode Island and to determine the incidence of poisoning deaths that are not reported to the regional poison center. DESIGN: The records of the state medical examiner were reviewed retrospectively for all toxin-related deaths from January 1986 through December 1989, and those deaths were compared with deaths recorded by the poison center during the same period. PARTICIPANTS: All patients whose deaths were due to a toxic substance and were recorded by either the state medical examiner's office or the regional poison center during the four-year study period. INTERVENTIONS: Age, sex, location of death, cause of death (both primary and secondary toxins were noted), manner of death (accidental, suicide, undetermined), and the circumstances surrounding the death were recorded for each victim. Medical examiner and poison center cases were matched and compared using a computerized data base program. RESULTS: During the study period, 369 deaths reported to the medical examiner were attributed to poisoning, while 45 poisoning deaths were recorded by the poison center. Of the deaths reported to the medical examiner, the most common reasons for lack of reporting to the poison center were that death occurred at a residence or patients were dead on arrival at the hospital and a toxin was not suspected until an autopsy was performed. Seventy nine other poisoning victims arrived at the hospital alive with a suspected toxic exposure and hypothetically could have been reported to the poison center but were not. CONCLUSION: The medical examiner's office represents a significant source of statistics regarding poisoning deaths, the majority of toxic deaths not reported to the poison center are dead on arrival, and using deaths as the dependent measure, the poison center may be underutilized in severe poisoning cases. PMID- 8239103 TI - Intramuscular methylprednisolone acetate for the prevention of relapse in acute asthma. AB - STUDY OBJECTIVE: To examine the hypothesis that an IM long-acting steroid injection will decrease the relapse rate of asthma patients. DESIGN: A randomized, single-blind, placebo-controlled trial. SETTING: The emergency department of an urban academic medical center. TYPE OF PARTICIPANTS: Acute asthma patients aged 18 to 45 years who were treated successfully in the ED. INTERVENTIONS: Participants received either IM saline placebo or 240 mg methylprednisolone acetate suspension at the time of discharge. MEASUREMENTS AND MAIN OUTCOME: Relapse, defined as the need to seek nonroutine medical care for asthma within seven days of discharge, was the main outcome measure. Of 70 patients entered, 56 (80%) completed follow-up, including 30 in the steroid group and 26 in the placebo group. The groups had similar characteristics and peak expiratory flow values on arrival and at discharge. Relapse occurred in two steroid patients (6.7%) and eight placebo recipients (30.8%) (P < .05). There was one death in the placebo group. Reported side effects were similar. CONCLUSION: IM long-acting steroids reduced the rate of relapse in patients with acute asthma. PMID- 8239104 TI - Changes in serum catecholamine levels during acute bronchospasm. AB - STUDY OBJECTIVES: Previous studies reported that norepinephrine levels increased during acute asthma, while epinephrine levels have remained at normal levels. The purpose of this study was to determine whether the lack of rise in epinephrine levels is due to an inherent defect in catecholamine secretion in asthmatics or is a result of an insufficient stimulus for epinephrine release. DESIGN: County owned, university-affiliated emergency department. SETTING: Prospective study. TYPE OF PARTICIPANTS: Patients more than 18 years old presenting to the ED with an acute exacerbation of asthma or chronic obstructive pulmonary disease (COPD). INTERVENTIONS: Initial phlebotomy and spirometry were followed by treatment with either albuterol or albuterol and glycopyrrolate. Asthmatic patients received steroids. Phlebotomy and spirometry were performed again after treatment. MEASUREMENTS AND MAIN RESULTS: There was no difference in epinephrine levels between patients with asthma and those with COPD either before or after treatment. Norepinephrine levels were higher in patients with COPD, both before and after treatment. There was only a weak correlation between epinephrine levels and pretreatment FEV1 for asthma but not for COPD. CONCLUSION: Moderate bronchospasm is not a major stimulus for epinephrine release in either acute asthma or COPD. PMID- 8239105 TI - Comparison of intermittent and continuously nebulized albuterol for treatment of asthma in an urban emergency department. AB - STUDY OBJECTIVE: To compare continuously nebulized albuterol with intermittent bolus nebulization of albuterol. DESIGN: Consecutive block enrollment in groups of ten to continuous or intermittent therapy. SETTING: Urban emergency department. TYPE OF PARTICIPANTS: Patients who presented to the ED with moderate to severe asthma and did not improve after one treatment with nebulized albuterol. INTERVENTIONS: All patients received an initial nebulized treatment with 2.5 mg albuterol followed by 125 mg solumedrol. Patients in the intermittent group received 2.5 mg nebulized albuterol at 30, 60, 90, and 120 minutes after the initial treatment. Patients in the continuous group received 10 mg albuterol nebulized in 70 mL over two hours. RESULTS: There was no difference between groups in age, sex, or initial peak expiratory flow rate (PEFR). Ninety-nine patients were included in the study (47 continuous and 52 intermittent). There was no statistically significant difference in PEFRs or admission rate between groups over the two-hour study period. One subgroup analysis was performed on patients with PEFRs on presentation to the ED of 200 L/min or less. Mean +/- SD baseline PEFR at presentation to the ED was 135 +/- 35 in the 35 patients in the continuous group and 137 +/- 45 in the 34 patients in the intermittent group). At 120 minutes, PEFR was 296 +/- 98 in the continuous group and 244 +/- 81 in the intermittent group (P = .01). Admission: discharge ratios for this subgroup analysis were 11:24 in the continuous group and 19:14 in the intermittent group (P = .03). Mean +/- SD heart rate in the subgroup analysis was 102 +/- 21 at baseline for the continuous group and 109 +/- 22 at baseline in the intermittent group. At 120 minutes, heart rate was 90 +/- 18 in the continuous group and 104 +/- 16 in the intermittent group (P = .002). CONCLUSIONS: Continuous nebulization offers no benefit over intermittent therapy in patients with an initial PEFR of more than 200 L/min. In PEFRs of 200 or less, continuous nebulization may decrease admission rate and improve PEFRs when compared with standard therapy. PMID- 8239106 TI - Continuous versus intermittent albuterol nebulization in the treatment of acute asthma. AB - STUDY OBJECTIVES: To compare bronchodilation, chronotropic effects, and side effects of the same dose of nebulized albuterol when given by either intermittent or continuous administration. DESIGN: A randomized assignment of patients to one of the two methods of albuterol nebulization. SETTING: Adult emergency department in an urban public hospital. TYPE OF PARTICIPANTS: Adult patients with acute exacerbations of asthma. INTERVENTIONS: Administration of 30 mg albuterol given over 110 minutes by either continuous or intermittent aerosolization. MEASUREMENTS: FEV1, forced vital capacity, heart rate, and systolic and diastolic blood pressures were measured immediately before treatment and then hourly for two hours. Side effects, symptoms, subsequent admission, and discharge also were noted. MAIN RESULTS: An overall significant decrease in heart rate was observed, indicating the lack of significant chronotropic effects with this dose of albuterol. Both treatments resulted in significant spirometric improvement without a significant treatment difference for the entire group. A difference, however, was found in the relative rates of FEV1 improvement with the two treatments depending on whether patients had an initial FEV1 less than or more than 50% predicted (P = .05). A secondary analysis on patients with an initial FEV1 less than percent predicted demonstrated a higher rate of percent predicted FEV1 increase with the continuously nebulized albuterol group (P = .03). CONCLUSION: This study demonstrates that albuterol can be given safely at this dose by either intermittent or continuous nebulization. Future studies should examine whether continuous nebulization has a reproducible advantage over intermittent nebulization in patients with more severe obstruction. PMID- 8239107 TI - Guidelines for the selective ordering of admission chest radiography in adult obstructive airway disease. AB - STUDY OBJECTIVE: To validate previously developed guidelines for the selective use of chest radiography in adults admitted for exacerbation of obstructive airway disease. DESIGN: Prospective, observational cohort study using criteria developed in a previous retrospective study. PARTICIPANTS: Unselected convenience sample of 128 adults with obstructive airway disease who did not respond to standard emergency department treatment and required admission. SETTING: Municipal hospital ED and inpatient medical service. INTERVENTIONS: Patients were categorized as "complicated" or "uncomplicated" according to previously developed criteria. Management was recorded as altered if the patient's physician answered the question, "Did the chest radiography alter your management of this patient?" affirmatively. RESULTS: Of 27 patients whose management was altered by the chest radiography, 26 were classified as complicated, for a sensitivity of 96% (95% confidence interval [CI], 81, 100). One of 44 admissions classified as uncomplicated had management altered by the chest radiography (negative predictive value, 98%, 95% CI, 88, 100). This chest radiography was later reread as normal. Classification as an uncomplicated patient with obstructive airway disease was strongly associated with either a normal chest radiography or a radiographic finding that was clinically unimportant (P = .0002). CONCLUSION: Patients with acute exacerbation of obstructive airway disease who are otherwise uncomplicated do not benefit from routine admission chest radiography. The use of this simple clinical strategy would safely reduce the number of chest radiographs by about one-third in this and similar patient populations, decreasing both health care costs and exposure to ionizing radiation. PMID- 8239108 TI - Data, epidemiology, and the future strength of emergency medicine. PMID- 8239109 TI - Are activated charcoal-N-acetylcysteine interactions of clinical significance? PMID- 8239110 TI - Post-dural-puncture headache: characteristics, management, and prevention. AB - Post-dural-puncture headache is a common complication after both diagnostic and therapeutic procedures. Prompt recognition of the clinical syndrome, followed by supportive and corrective actions, can decrease the morbidity in those afflicted. The most useful preventive measure during dural puncture in adult patients is the use of smaller cannulae with the bevel aimed parallel to the long axis of the spine. PMID- 8239111 TI - Recent developments in our understanding of sepsis: evaluation of anti-endotoxin antibodies and biological response modifiers. AB - Sepsis and septic shock are among the most frequent life-threatening infectious disease problems encountered in emergency medicine practice. This review summarizes the extensive research into the pathophysiology of sepsis, with emphasis on Gram-negative infection. Particular reference is given to the exogenous and endogenous mediators involved in the sepsis cascade. It also critically evaluates new preparations developed to blunt the actions of the exogenous and endogenous mediators responsible for the clinical manifestations comprising this syndrome. Clinical signs likely to be associated with Gram negative infection are also reviewed, and guidelines are considered for the potential use of newly developed anti-endotoxin antibodies and other biological response modifiers in the treatment of patients with Gram-negative sepsis. PMID- 8239112 TI - When does a guideline become a standard? The new American Society of Anesthesiologists guidelines give us a clue. AB - So when does a guideline become a standard? The answer is when an inexpensive, reliable device comes onto the market, the technology and concept of which have already been adopted by a group who specialize in the concept of intubation- anesthesiologists. A guideline becomes a standard of care when the device behind the guideline is available and readily usable as a practical matter by members of other medical specialties who have cause and reason to consider its use. By the time a piece of technology is recognized by legislators or insurance company underwriting departments as a necessary tool in the arsenal of a group of medical professionals, essentially as a standard of care, it is too late in the sense that the damage will already have been done because the malpractice suits that motivated the legislative or underwriting change already will be inescapably present. The mandation of a standard of care by legislative or insurance underwriting fiat certainly ensures the existence of the standard and will serve to require the use of the procedures, techniques, or technology so mandated. Before the mandation by law, however, the profession must recognize the matriculation of the idea to the guideline to the standard. Waiting for adoption of the standard by law or insurance company underwriting departments gives the plaintiff's trial bar "an edge," in that they are able to assert that a standard exists that the physicians are ignoring.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239113 TI - Methcathinone: a Russian designer amphetamine infiltrates the rural midwest. AB - Four cases of methcathinone toxicity in the United States are presented and discussed. This drug is the latest in a series of "designer" amphetamines. Prominent findings of acute toxicity include hallucinations, fever, and tachycardia followed by periods of bradycardia and moderate hypotension as acute symptoms resolve. Chronic binge use may result in development of paranoid psychosis and brief periods of withdrawal tremors. Our current understanding of methcathinone effects and the Russian experience with it are discussed. Our recommendations for treatment of acute toxicity are based on well established guidelines for managing the toxicity of similar amphetamine-like drugs. PMID- 8239114 TI - Treatment of foxglove extract poisoning with digoxin-specific Fab fragments. AB - A 22-year-old man presented to our emergency department after an intentional overdose of a homemade foxglove extract. Clinical symptoms with symptomatic bradyarrhythmia and ECG changes were consistent with cardiac glycoside poisoning. Treatment with digoxin-specific Fab fragments resulted in transient clinical and ECG improvement. Serum immunoassay demonstrated a digitoxin-like glycoside. The serum levels showed no evidence of altered elimination or distribution with Fab therapy despite temporary improvements in the clinical course. The use of Fab did not result in a shortened clinical course in this episode of foxglove poisoning, as one would expect in the setting of commercial glycoside product poisoning. PMID- 8239115 TI - Use of sodium polystyrene sulfonate in a lithium overdose. AB - A 23-year-old woman with an acute-on-chronic lithium overdose received multiple oral doses of sodium polystyrene sulfonate totaling 150 g over a 24-hour period. During the 33 hours after the institution of therapy, the serum lithium level decreased from 4.20 to 0.68 mEq/L. The calculated serum lithium elimination half life of 12 hours is significantly shorter than that previously noted in other similar overdoses, and the patient suffered no adverse effects of therapy. Multiple-dose sodium polystyrene sulfonate may be useful in lowering the serum lithium level of select patients with acute lithium overdoses but is not a substitute for hemodialysis in severely ill patients. PMID- 8239116 TI - Inhalational and percutaneous methanol toxicity in two firefighters. AB - We present two cases of adult inhalational and percutaneous methanol toxicity resulting from transient exposure to vaporized methanol. Both patients complained only of a mild headache at the time of the emergency department evaluation and had normal physical examinations, normal anion gaps, and peak methanol levels of 23 and 16 mg/dL, respectively. Emergency physicians should recognize the potential for toxic transcutaneous absorption of methanol. Because of the varying relationship between clinical symptoms, physical examination findings, and anion gap values to potentially toxic methanol exposures, acquisition of empiric serum methanol levels appears warranted in appropriate situations. PMID- 8239117 TI - Methanol inhalation toxicity. AB - Significant toxicity can result from intentional methanol inhalation. We report seven cases, involving four patients, of intentional inhalation of CARB-MEDIC carburetor cleaner containing toluene (43.8%), methanol (23.2%), methylene chloride (20.5%), and propane (12.5%). Patients arrived at the emergency department with central nervous system depression, nausea, vomiting, shortness of breath, photophobia, and/or decreased visual acuity. Treatment included correction of acidosis, leucovorin and/or folic acid, ethanol infusions, and supportive care. Hemodialysis was necessary in three cases. Measured blood methanol levels ranged from 50.4 to 128.6 mg/dL. Blood formic acid levels were 120, 193, and 480 micrograms/mL, respectively, in three patients. Ophthalmic examinations revealed hyperemic discs and decreased visual acuity in one patient. One individual was found pulseless with several CARB-MEDIC cans nearby. Attempts at revival were unsuccessful. Clinicians should be aware that significant blood methanol and formic acid levels may occur after inhalation of methanol. PMID- 8239118 TI - Fetal monitoring with adenosine administration. PMID- 8239119 TI - Teaching optimal paddle position for defibrillation. PMID- 8239120 TI - Infrared emission detection tympanic thermometry. PMID- 8239121 TI - Desipramine and amantadine causing false-positive urine test for amphetamine. PMID- 8239122 TI - Antigen-capture enzyme immunoassay for detection of avian influenza virus in turkeys. AB - A double-antibody sandwich ELISA (DAS-ELISA) was developed for detection of avian influenza virus (AIV) antigen. A monoclonal antibody to the viral nucleoprotein (NP) was used to coat the ELISA plates. A direct DAS-ELISA and an indirect DAS ELISA were evaluated. In the direct DAS-ELISA, monoclonal antibody to the AIV NP conjugated with horseradish peroxidase was used. The direct DAS-ELISA was evaluated for its sensitivity to detect purified NP; this procedure detected as little as 0.1 ng. In the indirect DAS-ELISA, rabbit NP antibody and horseradish peroxidase-conjugated goat anti-rabbit immunoglobin were used as primary and secondary antibodies, respectively. The indirect DAS-ELISA was evaluated for its ability to detect the AIV antigen in tracheal and cloacal specimens from turkeys inoculated with AIV. Results of indirect DAS-ELISA were compared with those of conventional virus isolation. Percentage agreement between indirect DAS-ELISA and virus isolation in AIV-positive samples was found to be 76.1% and, in AIV negative samples, it was found to be 82.1%. These results indicate that the DAS ELISA might be a viable alternative to virus isolation because of its rapidity, compared with virus isolation. PMID- 8239123 TI - Enzyme-linked immunosorbent assay for serologic detection of Salmonella dublin carriers on a large dairy. AB - Cows and calves from a 1,600-cow drylot dairy were screened for IgG antibodies to Salmonella dublin lipopolysaccharide (LPS), using an indirect ELISA. The ELISA was performed on milk samples from lactating cows and on sera from nonlactating cows and calves. Fecal samples were collected from calves and nonlactating cows for culture of Salmonella spp. All seropositive cattle were retested by culture and ELISA 5 times at monthly intervals or until antibody concentration decreased. None of the cattle remained culture-positive and seronegative. Prior to and during the sample collection period, approximately 30% of calves < 8 weeks old died of S dublin infection. Vaccination of cows with a killed S dublin/S typhimurium vaccine at cessation of lactation was a routine management practice. The ELISA-determined IgG response to vaccination had decreased by 50 days after vaccination. Eight cows and 5 calves that maintained a high serologic response to S dublin were purchased and moved to a research facility for 6 months of intensive monitoring. Lactating cows were milked twice daily, and culture of milk and feces for Salmonella spp was performed 5 times/wk. Serum IgG antibodies to S dublin LPS were measured weekly, using ELISA. At the end of 6 months, all 13 cattle were necropsied and tissues were obtained for culture of Salmonella spp. All 8 cows and 5 calves maintained persistently high ELISA titer for the 6 months of testing, and shed S dublin in the milk and/or feces during the same period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239124 TI - Use of survival analysis to compare cull rates between bovine leukemia virus seropositive and seronegative dairy cows. AB - Bovine leukemia virus (BLV) infection and culling of cows in a commercial dairy herd were evaluated to determine whether a relation existed between the 2 factors. Cattle from the study population, a Holstein dairy herd consisting of approximately 400 milking cows, were tested for antibodies to BLV, using the agar gel immunodiffusion test, semiannually for 2 years, annually for 2 years, and when cattle were culled. Complete records of BLV test results were available for 849 (79%) of the 1,078 cattle that had at least 1 test during the study period. Using the Cox hazard model, the cull hazard rates (culls/cow-months) were greater for BLV seropositive cows than for seronegative cows > 36 months old. Hence, among older dairy cows, BLV-infected cows were culled prematurely, compared with uninfected cows. PMID- 8239125 TI - Serum tumor necrosis factor alpha concentrations and clinical abnormalities in colostrum-fed and colostrum-deprived neonatal foals given endotoxin. AB - We examined the effect of infusion of lipopolysaccharide (LPS) on serum tumor necrosis factor alpha (TNF alpha) concentration and clinical attitude in 2- 3-day old colostrum-fed (CF) and colostrum-deprived (CD) foals. Eleven CF and 8 CD neonatal foals were given a bolus i.v. infusion of Escherichia coli O55:B5 lipopolysaccharide (0.5 microgram/kg of body weight) in sterile saline (0.9% NaCl) solution. Four CF and 2 CD foals were given saline solution alone. Serum IgG concentration and serum anti-LPS IgG(T) antibody titer were determined for each foal prior to infusion. A depression index was used to score clinical abnormalities. Serum TNF alpha concentration was estimated by use of an in vitro cytotoxicity bioassay that used WEHI 164 clone 13 cells as targets. The cytotoxic serum factor was identified as TNF alpha by immunoprecipitation with caprine antisera raised against the 15 NH2-terminal amino acids of human TNF alpha. Tumor necrosis factor alpha was not detected in any preinfusion serum samples nor in any samples from foals given saline solution alone. Serum TNF alpha concentration increased in all LPS-infused foals and peaked between 60 and 90 minutes after infusion. Serum TNF alpha concentrations, expressed as mean percentage of peak serum TNF alpha concentration, persisted longer in CD foals given LPS than in CF foals given LPS. All LPS-infused foals displayed clinical signs of endotoxemia, but mean depression index scores of the CF and CD foals given LPS were not significantly different at any time. Serum TNF alpha concentrations were correlated with depression index scores in both LPS-infused groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239126 TI - Serum interleukin-6 concentrations in endotoxin-infused neonatal foals. AB - Serum interleukin-6 (IL-6) concentration was measured in 11 colostrum-fed (CF) and 8 colostrum-deprived (CD) 2- to 3-day-old foals after foals were infused with lipopolysaccharide (LPS; Escherichia coli O55:B5 endotoxin, 0.5 microgram/kg of body weight in sterile saline [0.9% NaCl] solution). Four CF and 2 CD foals were given saline solution alone. Serum IL-6 concentration was estimated by use of an in vitro proliferative bioassay, using the IL-6 dependent B.13.29 clone 9 cells. Interleukin-6 concentration increased in all LPS-infused foals, and geometric mean serum IL-6 concentration was significantly higher in CF than CD foals 30 and 90 minutes after infusion. Both LPS-infused groups had multiple spikes of mean IL 6 concentration that peaked at 120 minutes in CF foals and 150 minutes in CD foals. Results indicated that IL-6 is produced in neonatal foals in response to LPS infusion. Furthermore, colostrum deprivation resulted in longer times to peak mean serum IL-6 concentration and tended to reduce serum IL-6 concentration in neonatal foals. PMID- 8239127 TI - Enzyme-linked immunosorbent assay and flow cytometric methods to screen hybridoma culture supernatants for antibodies to bovine neutrophil surface antigens, and monoclonal antibody production and characterization. AB - Enzyme-linked immunosorbent assay and flow cytometric methods to screen hybridoma culture supernatants for antibodies to bovine neutrophils (surface antigen specific) were optimized. Sensitivity of the 2 methods was compared. A panel of 14 murine monoclonal antibodies (MAB) to surface antigens of bovine polymorphonuclear neutrophilic leukocytes (neutrophils) was produced by hybridoma technology, and their isotypes were determined by whole-cell ELISA. Monoclonal antibody reactivity with neutrophils, eosinophils, and lymphocytes isolated on phosphate-buffered saline solution and on Ficoll-sodium diatrizoate were compared. Biochemical characterization of antigens recognized by MAB was performed by immunoblot analysis. Neutrophil plasma membranes were isolated on sucrose gradients (20, 32, and 50%) and purified for polypeptide characterization. Neutrophil surface proteins were characterized by external labeling with 125I. The flow cytometric method was proven to be more sensitive and rapid than ELISA to screen hybridoma supernatants. This method allowed light scatter gating of live neutrophil populations for analysis, which eliminated nonspecific binding of antibodies to contaminating cells and dead neutrophils. The optimal conditions for flow cytometric analyses were 5 x 10(5) neutrophils and 1 micrograms of fluorescein-labeled F(ab')2/assay as the second antibody. The optimal conditions for hybridoma screening by ELISA were neutrophil concentration of 2.5 x 10(5)/well, using a 96-well polystyrene microtitration plate as solid support, and 2,2'-azino-di[3-ethyl-benzthiazoline sulfonate (6)] with H2O2 as the chromatogenic substrate. Tissue culture plates as solid support and 3,3', 5,5' tetramethyl benzidine, with H2O2 as the chromogenic substrate, were equally as sensitive. Panel MAB reacted differently with neutrophils, eosinophils, and lymphocytes. Isolation of these cells from blood on Ficoll-sodium diatrizoate generally did not alter MAB reactivity. Coomassie blue-stained gels of neutrophil plasma membrane proteins contained about 25 polypeptide bands, 13 of which were major bands. Autoradiography revealed about 11 surface proteins, 5 of which were heavily labeled with 125I. Monoclonal antibody S7G8 identified a 65-kd protein and MAB S8G10 identified 65- and 70-kd proteins. On the basis of molecular weight, MAB S7G8 and S8G10 are comparable to human CD15, CD16, and CD64 molecules. The MAB generated in this study are potential candidates to discern bovine neutrophil function and heterogeneity. PMID- 8239128 TI - Production of monoclonal antibodies specific for antigens derived from tissue of chinook salmon (Oncorhynchus tshawytscha) affected with plasmacytoid leukemia. AB - Two distinct monoclonal antibodies (MAB) were prepared for testing with kidney, spleen, and retrobulbar tissue imprints made from chinook salmon (Oncorhynchus tshawytscha) affected with plasmacytoid leukemia. (PL). Hybridomas were prepared from mice immunized with whole and lysed cells purified from renal or retrobulbar PL-positive tissues, which had been obtained from naturally and experimentally infected fish from British Columbia, Canada. The MAB reacted with at least 4 morphologically different cell types; fluorescence was associated with the plasma membrane and cytoplasm. The MAB also reacted with kidney imprints made from chinook salmon affected with a PL-like lymphoproliferative disease in California, indicating that these 2 diseases might be caused by a similar agent. The MAB did not react with any of the kidney or spleen imprints made from wild chinook salmon collected from a river in Ontario, Canada. PMID- 8239129 TI - Interspecific conjugal transfer of antibiotic resistance among staphylococci isolated from the bovine mammary gland. AB - In vitro transferability of penicillin, streptomycin, tetracycline, and erythromycin resistance from coagulase-negative staphylococci to Staphylococcus aureus and among the former species of bovine mammary gland origin was examined by bacterial mating on filters and by mixed-culture matings in broth and in skim milk. One hundred twenty-six (42 each on filter, in broth, and in skim milk) matings were performed among 37 isolates of different Staphylococcus species. Transfer of resistance to penicillin, tetracycline, or erythromycin was not detected. Of 51 matings performed to determine streptomycin-resistance transfer, 9 (3 each on filters, in broth, and skim milk) were successful. Nine strains representing 3 species of coagulase-negative staphylococci were tested as prospective donors of streptomycin resistance. Of these, 2 strains could transfer streptomycin resistance. A double-resistant donor, S hominis, not only transferred its streptomycin resistance to an S chromogenes strain lacking resistance, but also to an S aureus strain already carrying penicillin and tetracycline resistance. The transfer of streptomycin resistance from the donor S hominis, harboring 2 plasmids, to a plasmidless S chromogenes recipient strain was associated with apparent acquisition of the smaller plasmid of the donor by the recipient. The single-resistant donor, S epidermidis 681A, transferred streptomycin resistance to a tetracycline-resistant S aureus recipient. This strain however, failed to transfer its streptomycin resistance to another S aureus, 2 S hyicus, and 1 S xylosus recipient. Frequency of transfer of streptomycin resistance ranged from 1.1 x 10(-5) to 1 x 10(-4). When transfer of resistance was successful, attempts were made to characterize the transfer process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239130 TI - Isolation of Mycobacterium paratuberculosis from mononuclear cells in tissues, blood, and mammary glands of cows with advanced paratuberculosis. AB - Seven mature dairy cows from 6 herds were obtained with history, clinical signs of disease, and laboratory findings suggestive of advanced paratuberculosis. A surgically implanted collection chamber was used to obtain peripheral tissue fluid. Blood, mammary gland flush fluid, and collection chamber flush fluid (CCFF) samples were obtained 6 times over a 2-week period from each cow. Mononuclear cell-rich portions of these fluids obtained by gradient centrifugation were submitted for bacteriologic culture of Mycobacterium paratuberculosis and for total and differential cell counts. Bacteriologic culture of feces for M paratuberculosis and complete necropsy performed on each cow at the conclusion of the study confirmed the diagnosis of paratuberculosis. Numbers of tissue macrophages obtained from CCFF samples were lower than expected. Mean (+/- SD) differential count of tissue macrophages collected from CCFF was 65.57 (+/- 23.39). Mean calculated tissue macrophages (total cell count x differential count) collected from CCFF samples was 623.1 (+/- 784.55) cells/microliters. Mycobacterium paratuberculosis was isolated from 1 of 42 (2.4%) collections of mononuclear cell-rich portions of plasma and from 2 of 42 (4.8%) CCFF samples. Mycobacterium paratuberculosis was not isolated from any collections of mammary gland flush fluid. The collection and processing techniques used in this study did not enhance detection of M paratuberculosis infection in cows with advanced paratuberculosis, beyond that of ileocecal lymph node biopsy or fecal culture. PMID- 8239131 TI - Enterotoxigenic, verotoxigenic, and necrotoxigenic Escherichia coli isolated from cattle in Spain. AB - To assess the role of enterotoxigenic (ETEC), verotoxigenic (VTEC), and necrotoxigenic (NTEC) Escherichia coli in cattle with diarrhea, 1,524 colonies of E coli isolated from 197 calves with diarrhea and from 112 healthy controls were investigated for production of heat-labile and heat-stable enterotoxins, verotoxins, and cytotoxic necrotizing factors (CNF1 and CNF2). The ETEC were isolated from only 2 (1%) calves with diarrhea and from 5 (4%) healthy controls. In contrast, VTEC and NTEC that produced CNF2 were frequently identified. The VTEC were isolated from 18 (9%) calves with diarrhea and from 21 (19%) healthy cattle (P < 0.05), whereas NTEC that produced CNF2 were detected in 39 (20%) ill calves and in 38 (34%) controls (P < 0.01). Therefore, VTEC and NTEC that produced CNF2 were isolated significantly more frequently from healthy than diseased calves. Serogroups to which VTEC belonged differed considerably from the O groups involved with NTEC. Although, VTEC belonged to 18 serogroups, only 4 (O26, O103, O113, and O157) accounted for 56% (25 of 45) of verotoxigenic strains. The NTEC that produced CNF2 belonged to 26 serogroups; however, 64% (69 of 108) were from 6 serogroups (O1, O3, O15, O55, O88, and O123). Our results are compatible with cattle being a reservoir of VTEC that are pathogenic for human beings and with ETEC being an unusual cause of bovine colibacillosis in Galicia (northwestern Spain). Furthermore, results of this study indicate that VTEC and NTEC that produced CNF2 may be part of the normal intestinal flora of cattle. PMID- 8239132 TI - Prevalence of bluetongue virus expression in leukocytes from experimentally infected ruminants. AB - Replication of bluetongue virus (BTV) in leukocytes from the blood of sheep, cattle, elk, and mule deer inoculated with BTV serotype 10 or 17 was assessed by immunocytochemical staining and dot blot northern hybridization to determine if differences in the prevalence of infection in this blood fraction might account for the differences in clinical disease among these species. Viremia was confirmed by virus isolation in all inoculated animals. Analysis of leukocytes with monoclonal antibodies specific for BTV proteins revealed low numbers of infected leukocytes in only 2 sheep 8 days after inoculation with BTV serotype 10. Most of the cells expressing BTV were identified morphologically as monocytes; approximately 10% of infected cells were lymphocytes. Bluetongue virus was not detected by use of dot-blot hybridization on samples of blood. Our results suggest that differential infection of leukocytes does not account for the pronounced differences in clinical signs and pathologic changes among ruminants. PMID- 8239133 TI - In situ hybridization of virulent canine distemper virus in brain tissue, using digoxigenin-labeled probes. AB - Only a few hybridization experiments have been performed for detection of canine distemper virus (CDV) nucleic acid sequences in tissue cultures and in various tissues. Those published studies used probes derived from tissue culture-adapted CDV, and hybridization signals were not obtained in the CNS tissue, although infective CDV and viral antigen were detectable in this tissue. We developed probes complementary to virulent CDV and were able to detect viral RNA not only in primary brain cell cultures, but also in brain tissues, by use of in situ hybridization. Sensitivity of the test at least equaled that of immunohistochemistry. We applied digoxigenin-labeled, strand-specific RNA probes complementary to the nucleoprotein-coding viral nucleic acid sequence. Our results indicate that to detect CDV nucleic acid sequences in brain tissues, it is essential to use probes derived from the virulent virus. PMID- 8239134 TI - Purification of surface-exposed integral outer membrane proteins of Actinobacillus pleuropneumoniae and their role in opsonophagocytosis. AB - Previously identified 39-, 50-, and 76-kd integral outer membrane proteins (IOMP) of Actinobacillus pleuropneumoniae, a respiratory tract pathogen, were separated by electroelution of sodium dodecyl sulfate-polyacrylamide gel electrophoresis obtained fragments and their role in opsonophagocytosis by porcine leukocytes was investigated by flow cytometry of fluorescein-labeled A pleuropneumoniae. Using specific antisera, immunoblot analysis indicated that the 3 proteins were antigenically distinct. Antibodies against each IOMP have an important role as opsonins for phagocytosis by porcine leukocytes. The effect of using a combination of all 3 of the specific antisera was minimal. Antiserum absorbed against intact A pleuropneumoniae and Escherichia coli organisms indicated that the antibodies to the 39-, 50-, and 76-kd IOMP were specific for A pleuropneumoniae antigens. Nonheat-treated antiserum did not increase phagocytosis, compared with heat-inactivated antiserum, indicating that complement may not have a major role in opsonization of A pleuropneumoniae. PMID- 8239135 TI - Increase of mannose residues, as Salmonella typhimurium-adhering factor, on the cecal mucosa of germ-free chickens infected with Eimeria tenella. AB - To study increase of the Salmonella population in the cecum of chickens infected with Eimeria tenella, quantitative changes in mannose residues on the cecal mucosa were investigated. Inhibition of S typhimurium adherence to the cecum by a 2% carbohydrate (D-mannose, D-galactose, L-fucose, alpha-methyl-D-glucoside) in phosphate-buffered saline solution was examined. Only D-mannose had inhibitory effects. Whereas, D-galactose had somewhat enhancing effects on adherence of S typhimurium to the cecal mucosa of uninfected germ-free chickens. In infected and uninfected chickens, D-mannose inhibited adherence of S typhimurium. D-Mannose significantly (P < 0.05) increased adherence of Bacteroides sp. In infected and uninfected chickens, D-mannose did not have any effect on adherence of Clostridium perfringens and Bifidobacterium thermophilum. Under microscopic observation, only concanavalin A and Lens culinaris agglutinin, of 8 lectins examined, were recognized as lectin-positive staining lines or spots in the cecal mucosa, indicating presence of mannose residues on the cecal mucosa. In E tenella infected chickens, lectin-positive staining was seen strongly on the coarse surface of damaged cells and at the bottom of the crypts. These results indicate that coccidial infection may induce increase of mannose residues on the intestinal surface and allow adhesion of more salmonellae to the intestine. PMID- 8239136 TI - Effect of estradiol administration and subsequent nonfeeding on liver estrogen receptor, serum apolipoprotein B-100, and serum triglycerides concentrations in steers. AB - Estradiol was administered to 3 steers (0.12 mg/kg of body weight/d for 14 consecutive days), followed by 2 days of nonfeeding (starvation). During estradiol administration, liver nuclear estrogen receptor and serum apolipoprotein B-100 (apoB-100), as well as serum triglycerides concentrations were increased, compared with values before administration. Starvation, together with interruption of estradiol administration, resulted in rapid decreases of the receptor, serum apoB-100, and serum triglycerides concentrations, and increase of nonesterified fatty acids concentration. Of the 3 steers, 2 had higher liver triglyceride content, compared with values before treatment. In the control group (3 steers that received vehicle alone, then starved similarly), these concentrations, except for serum nonesterified fatty acids and triglycerides concentrations after starvation, were not changed. In another experiment, serum apoB-100 concentration in dairy cows was significantly (P < 0.05) lower at parturition than values before and after parturition. These results indicate that estradiol may be involved in development of fatty liver in cattle. PMID- 8239137 TI - Antinociceptive effects of combining low doses of neuroleptic drugs and fentanyl in sheep. AB - Effects of low doses of the neuroleptic drugs droperidol and zuclopenthixol, combined with a subanalgesic dose of the opioid mu-agonist, fentanyl, on mechanical nociceptive thresholds were evaluated in sheep. Intravenously administered droperidol (5 micrograms/kg of body weight) did not induce any change in the nociceptive thresholds when administered alone, but caused marked increase in threshold responses when combined with a subanalgesic dose of fentanyl (5 micrograms/kg). Similarly, a combination of i.v. administered zuclopenthixol (100 micrograms/kg) and fentanyl induced significant (P < 0.05) antinociceptive effects, whereas zuclopenthixol administered i.v. alone had no effect on the threshold responses. Intrathecal administration of a low dose of droperidol (5-micrograms total dose) combined with i.v. administered fentanyl also increased mechanical thresholds significantly (P < 0.05). These results indicate that interactions exist between dopaminergic and opioid systems in the processing of nociceptive information and that these effects may, at least partially, be mediated spinally. PMID- 8239138 TI - Effect of preanesthetic medication on ease of endoscopic intubation of the duodenum in anesthetized dogs. AB - The effects of preanesthetic medication on ease of duodenal endoscopic intubation in dogs was evaluated. One of 12 combinations of preanesthetic medications (using atropine, glycopyrrolate, morphine, meperidine, acepromazine, and 0.9% NaCl solution) was administered IM to each of 12 dogs in a trial. Twelve endoscopic trials were performed so that each dog received each treatment combination once. Anesthesia was induced with thiamylal administered IV and maintained with halothane vaporized in oxygen. Electrocardiographic recordings, indirect blood pressure measurements, end-tidal carbon dioxide partial pressures, and halothane concentrations were monitored during the anesthetic period. The ease with which the fiberoptic endoscope was passed into the proximal portion of the duodenum was qualitatively score on the basis of time and maneuvering effort. None of the preanesthetic combinations made intubation of the duodenum significantly easier than that with 0.9% NaCl solution (control). Only the combination of morphine and atropine induced gastro-pyloric conditions that resulted in significantly higher (more difficult) endoscopic scores than those after preanesthetic medication with 0.9% NaCl solution. PMID- 8239139 TI - In vitro antibacterial activity of cefoxitin and cefotetan and pharmacokinetics in dogs. AB - The susceptibility of 50 clinical Escherichia coli isolates to various antibacterials, including cefoxitin and cefotetan was ascertained, and the minimal inhibitory concentration (MIC) of cefoxitin and cefotetan for each of these isolates was determined. The pharmacokinetics of cefoxitin and cefotetan after a single i.v. or SC injection (30 mg/kg of body weight) were determined in 4 dogs. Of the 50 E coli isolates, 98% were susceptible in vitro to cefotetan, 90% were susceptible to cefoxitin, and 88% were susceptible to gentamicin. The MIC that would inhibit the growth of 90% of the E coli isolates (MIC90) was 0.25 microgram/ml for cefotetan and 4 micrograms/ml for cefoxitin. Plasma cefotetan concentrations remained above MIC90 for (mean +/- SD) 8.2 +/- 1.72 hours and 13.52 +/- 0.28 hours after i.v. and SC administration, respectively. Plasma cefoxitin concentrations remained above MIC90 for (mean +/- SD) 5.37 +/- 1.18 hours and 7.95 +/- 0.71 hours after i.v. and SC administration, respectively. We concluded that cefotetan was superior to cefoxitin in activity against E coli in vitro. We recommend that cefotetan be given at a dosage of 30 mg/kg, i.v., every 8 hours, or SC, every 12 hours. PMID- 8239140 TI - Effect of furosemide and weight carriage on energetic responses of horses to incremental exertion. AB - The effect of furosemide-induced weight loss on the energetic responses of horses to running was examined in a 3-way crossover study. Eight 2- to 3-year-old Standardbred mares received, in random order, 10 ml of saline solution 4 hours before running on a treadmill (control trial, C); or, during 2 trials, 1 mg of furosemide/kg of body weight, i.v., 4 hours before running. During one of the trials when the horses received furosemide, they carried weight equal to that lost over the 3.75 hours after furosemide administration while running (furosemide-loaded, FL), and during the other trial they did not carry weight equal to that lost after furosemide administration (furosemide-unloaded, FU). Horses performed an incremental exercise test on a treadmill during which rates of oxygen consumption (VO2) and carbon dioxide production (VCO2) were measured, respiratory exchange ratio was calculated, and blood samples were collected for determination of mixed venous plasma lactate concentration and arterial and mixed venous oxygen saturation. Furosemide treatment caused significantly (P < 0.001) greater weight loss than did saline administration; mean +/- SEM weight loss (exclusive of fecal loss) was 1.6, 8.8, and 10.2 kg (SEM = 2.0) for C, FL, and FU trials, respectively. The speed at which peak VO2 was achieved was 9.31, 9.56, and 9.50 (SEM = 0.16) m/s, respectively, time to fatigue was 547, 544, and 553 (SEM = 26) seconds, respectively, and the highest speed attained was 10.3, 10.2, and 10.2 (SEM = 0.2) m/s, respectively. Mean peak rate of oxygen consumption was 130.7, 129.6, and 129.6 (SEM = 1.9) ml/min/kg, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239141 TI - Capacitation-like membrane changes and prolonged viability in vitro of equine spermatozoa cultured with uterine tube epithelial cells. AB - Reliable capacitation of equine spermatozoa has been a major obstacle in the development of equine in vitro fertilization. Experiments were done to compare in vitro capacitation of equine spermatozoa by use of heparin/caffeine, calcium ionophore, uterine tube epithelial cell (UTEC)-conditioned medium, and direct culturing of spermatozoa with UTEC (coculturing). Capacitation-like changes, as determined by chlortetracycline membrane staining patterns, developed with UTEC conditioned medium and coculturing, equivalent to that with calcium ionophore. Both of these treatments induced more (P < 0.05) capacitation-like changes than did the control, a modified Tyrode's medium. More (P < 0.05) spermatozoa were viable after 24 hours of UTEC coculturing than in the control incubation. PMID- 8239142 TI - Comparative efficacy of flunixin, ketoprofen, and ketorolac for treating endotoxemic neonatal calves. AB - Saline (0.9% NaCl) solution or 1 of 3 nonsteroidal anti-inflammatory drugs (NSAID) was administered i.v. to 5 neonatal calves 15 minutes after the start of a 3-hour i.v. infusion of Escherichia coli lipopolysaccharide (LPS; 2 micrograms/kg/h). Four additional calves were given a 3-hour i.v. infusion of saline solution alone. Clinical attitude, mean arterial blood pressure, PCV, WBC, and plasma lactate, glucose, and eicosanoid concentrations (thromboxane B2, 6 keto-PGF1 alpha) were monitored for 12 hours. Flunixin meglumine (1.1 mg/kg of body weight, i.v.), ketoprofen (2.2 mg/kg, i.v.), and ketorolac tromethamine (1.1 mg/kg, i.v.) each ameliorated the clinical signs of endotoxemia and LPS-induced lacticemia, but failed to significantly alter the degree of leukopenia or hypoglycemia associated with infusion of LPS. Although the 3 NSAID prevented eicosanoid production, they provided only partial protection against LPS-induced hypotension. Each NSAID modified the response to LPS, but none was clearly superior to the others in modulating the clinical signs or physiologic alterations induced by infusion of LPS in neonatal calves. PMID- 8239143 TI - Comparison of flunixin, prednisolone, dimethyl sulfoxide, and a lazaroid (U74389F) for treating endotoxemic neonatal calves. AB - Saline (0.9% NaCl) solution, flunixin meglumine (1.1 mg/kg), prednisolone sodium succinate (1.1 mg/kg), U74389F (1.5 mg/kg), and dimethyl sulfoxide (0.5 g/kg) were each administered i.v. to 5 neonatal calves 15 minutes after the start of a 3-hour infusion of Escherichia coli lipopolysaccharide (LPS; 2 micrograms/kg/hr). Four additional calves were given a 3-hour i.v. infusion of saline solution alone. Only flunixin significantly suppressed eicosanoid production and mitigated clinical signs associated with endotoxemia. Prednisolone provided partial protection against LPS-induced hypotension and lacticemia. Pronounced hypoglycemia and lacticemia were observed in U74389F-treated calves; LPS-induced hypotension and hypoglycemia were marked in dimethyl sulfoxide-treated calves. PMID- 8239144 TI - Effects of subject velocity on force plate-measured ground reaction forces in healthy greyhounds at the trot. AB - Force plate gait analysis was used to study the effects of subject velocity on ground reaction forces. Seven adult Greyhounds were trotted at 3 distinct velocities: 1.5 to 1.8 m/s, 2.1 to 2.4 m/s, and 2.7 to 3.0 m/s. Forelimb and hind limb peak vertical forces increased with increase in velocity (P < 0.05). Forelimb and hind limb vertical impulses decreased as velocity increased (P < 0.05). Significant variations were not observed for craniocaudal or mediolateral peak forces or impulses. It was concluded that velocity significantly (P < 0.05) influenced ground reaction forces and impulses, and must be controlled in experimental design. PMID- 8239145 TI - Effects of hylan on amphotericin-induced carpal lameness in equids. AB - In this double-blind study, the effectiveness of and dose response to intra articular administration of modified hyaluronan (hylan) was determined in an equine carpal lameness model over a 23-day period, using a computerized three dimensional motion analysis system, synovial fluid variables, and synovial histologic examination. In 24 clinically sound horses, baseline motion data was acquired from horses trotting at 4 m/s on a high-speed treadmill. Then, to induce lameness, 25 mg of amphotericin B in 5 ml of sterile water was injected into the left middle carpal joint of each horse every other day for 3 treatments. Phenybutazone (2.2 mg/kg of body weight, PO, once) and butorphanol tartrate (0.1 mg/kg, IM, q 6 h, for 36 hours) were used to control signs of discomfort. Horses were assigned at random to 4 equal groups and received intracarpal administration of either 1, 2, 4 ml of hylan (8 mg/ml), or 2 ml of balanced electrolyte solution (control). Intracarpal administration of amphotericin B caused significant (P < or = 0.01) increase in subjective lameness grades over the 2-week evaluation period, and hylan administration did not significantly (P < or = 0.01) change the subjective lameness grade. Lameness induction caused significant (P < or = 0.01) decrease in head and withers excursions during the lame forelimb support phase and significant (P < or = 0.05) increase in head and withers excursions during the sound forelimb support phase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239146 TI - Computer-assisted three-dimensional gait analysis of amphotericin-induced carpal lameness in horses. AB - Motion of 6 clinically sound horses trotting at a speed of 4 m/s on a treadmill was captured by video cameras before and 9, 16, and 23 days after amphotericin induced lameness to determine the quantitative variables of three-dimensional computer-assisted image analysis that objectively describe carpal lameness. Amphotericin-B was used to induce lameness, and phenylbutazone (2.2 mg/kg of body weight, PO, once) and butorphanol tartrate (0.1 mg/kg IM, q 6 h, to effect) were used to control discomfort. Four 60-Hz cameras were symmetrically placed around the treadmill to capture 6 seconds of images from retro-reflective spheres taped to the trotting horses. Images were transferred to a video-based digitizer and a computer work station, where 4 files of two-dimensional data were reduced to 1 file of three-dimensional data. The effect of lameness on motion analyzed was assessed by use of two-way ANOVA. Differences between means were assessed, using the Student-Newman-Keul's test (P < or = 0.05). Head and withers excursions, (dorsal vertical displacement of head and withers targets, respectively) during the sound forelimb support phase were increased significantly during all lameness measurement periods. Head excursion, but not withers excursion, during the lame forelimb support phase, was decreased significantly during all lameness measurement periods. Computer determinations of stride length swing phase, stance phase, forelimb abduction, and carpal and fetlock ranges of motion did not consistently characterize the lameness. It was concluded that three-dimensional computer-assisted image analysis could be used for objective lameness evaluation in horses and that head and withers excursions were the most consistent variables for assessing equine carpal lameness. PMID- 8239147 TI - Guidelines for animal surgery in research and teaching. AVMA Panel on Animal Surgery in Research and Teaching, and the ASLAP (American Society of Laboratory Animal Practitioners). AB - The direct or delegated authority for activities involving animals used in research and teaching programs lies with the attending veterinarian. These duties and responsibilities cannot be abrogated by others in the organization. The veterinarian is the single individual with the broadest range of knowledge of animals used in biomedical research, testing, and teaching. Federal regulations and policy require that the Institutional Animal Care and Use Committee (IA-CUC) ensure that the use of animals will be ethically justifiable and humane and that all individuals who perform surgery on animals will be appropriately qualified and trained. The attending veterinarian also has specific responsibilities under law and must take an active role in providing this assurance. In recognition of this, the Council on Research of the AVMA created a Panel on Animal Surgery in Research and Teaching to develop a report on guidelines for performing animal surgery in a research or teaching setting. Drafts of the panel report were sent through an extensive review process that included representative groups in veterinary medicine, human health, government, biomedical research, and animal welfare. The report of the panel was referred to the American Society of Laboratory Animal Practitioners (ASLAP). As an organization, the major focus of which is the role of the veterinarian in research using animals, it is appropriate that the ASLAP further refine this report into a set of guidelines for the research community. The ASLAP committee wishes to acknowledge the AVMA for recognizing the need for these guidelines and providing the financial support to convene the initial panel. This document was compiled with expectations for change in future revisions. Intended to complement other pertinent guidelines and regulations on animal surgery, these guidelines are designed to help individuals charged with the stewardship of animals used in research and teaching. PMID- 8239148 TI - The coming of age of a bronchial epithelial cell. PMID- 8239149 TI - Endothelin-1 in adult respiratory distress syndrome. AB - Endothelin-1 (ET-1), a potent vasoconstrictor peptide produced by endothelial cells and degraded predominantly in the pulmonary vasculature, has been implicated in the development of various organ dysfunctions. To determine the pathophysiologic role of ET-1 in adult respiratory distress syndrome (ARDS) and the impact of impaired lung function on transpulmonary peptide handling, we compared plasma levels and pulmonary ET-1 balance in 14 patients with ARDS and in seven healthy control subjects. To obtain comparable conditions in both groups, the ET-1 level was raised in the control group by exogenous infusion (0.4 pmol/kg/min) to 9.4 +/- 0.8 pmol/L. ARDS was accompanied by a hyperdynamic circulatory pattern with increased cardiac output and depressed total vascular resistance but, simultaneously, pulmonary hypertension. Venous ET-1 concentration was massively increased in ARDS (9.8 +/- 1.2 versus 2.1 +/- 0.2 pmol/L, p < 0.001). In control subjects, the lung cleared the major fraction of ET-1 (fractional extraction 43 +/- 8.8%, uptake 12.5 +/- 2.5 pmol/min). In contrast, in ARDS there was a pronounced pulmonary releases into the circulation (32.8 +/- 10.3 pmol/min). We conclude that ET-1 concentrations are elevated in ARDS as the result of both increased formation and decreased disposal. Lung failure affects not only gas exchange but also nonrespiratory, metabolic pulmonary functions. PMID- 8239150 TI - Oxidized glutathione is increased in the alveolar fluid of patients with the adult respiratory distress syndrome. AB - The adult respiratory distress syndrome (ARDS) is a devastating clinical illness characterized by refractory hypoxemia and high-permeability pulmonary edema. Reactive oxygen species may play a key role in the pathogenesis of the acute lung injury. Glutathione is a tripeptide that is able to react with and effectively neutralize oxidants, such as hydrogen peroxide. The present study found that the alveolar epithelial lining fluid of patients with ARDS was deficient in total glutathione compared with that of normal subjects (31.5 +/- 8.4 versus 651.0 +/- 103.1 microM, p = 0.0001) and patients with cardiogenic pulmonary edema (31.5 +/- 8.4 versus 154.1 +/- 52.4 microM, p = 0.001). In addition, a greater percentage of total glutathione was in the oxidized form in patients with ARDS compared with normal subjects (30.6 +/- 6.1 versus 6.4 +/- 2.9%, p = 0.03). This deficiency of reduced glutathione in the alveolar fluid may predispose these patients to enhanced lung cell injury. PMID- 8239151 TI - Endotoxin primes perfused rabbit lungs for enhanced vasoconstrictor response to staphylococcal alpha-toxin. AB - The major pore-forming exotoxin of Staphylococcus aureus, staphylococcal alpha toxin, causes thromboxane-mediated pulmonary hypertension and prostanoid independent protracted vascular leakage in perfused rabbit lungs. We asked whether lung responsiveness to the staphylococcal agent would be altered by a preceding period of endotoxin priming. Isolated rabbit lungs were perfused with Krebs-Henseleit buffer in the presence or absence of 100 ng/ml Salmonella abortus equii endotoxin for up to 5 h. The lipopolysaccharide exposure evoked the release of large quantities of tumor necrosis factor into the vascular and alveolar spaces but did not significantly alter pulmonary artery pressure, organ weight, or the repeatedly assessed capillary filtration coefficient (Kfc). Two and 4 h after endotoxin administration, alpha-toxin (10 to 30 ng/ml) was bolus-injected into the pulmonary artery. Toxin-evoked prostanoid generation (TxB2, 6-keto-PGF1 alpha) and pressor responses were markedly accelerated and enhanced in endotoxin primed lungs, both for the 2 h and the 4 h priming period. No significant influence of endotoxin was noted when applied simultaneously with alpha-toxin. Cyclooxygenase inhibition suppressed the alpha-toxin-evoked pressure rise in both endotoxin-primed and nonprimed lungs. Endotoxin priming did not influence the alpha-toxin-induced protracted increase in Kfc values, assessed in the presence of cyclooxygenase inhibition. We conclude that endotoxin primes rabbit lungs for enhanced prostanoid generation and pulmonary hypertension in response to S. aureus alpha-toxin. Such cooperativity of endotoxin priming and exotoxin triggering may be relevant in critically ill patients suffering from both endotoxemia and gram-positive sepsis. PMID- 8239152 TI - Aerosolized versus instilled exogenous surfactant in a nonuniform pattern of lung injury. AB - Previous studies have shown that the underlying patterns of lung injury influence subsequent responses to aerosolized exogenous surfactant. The purpose of this study was to compare aerosolized versus tracheally instilled surfactant in a nonuniform lung injury. Adult sheep underwent whole lung lavage with subsequent HCl instillation into the right middle lobe (RML) and lingula (LING) to create a nonuniform injury. Animals were treated with either nebulized surfactant (Neb.Surf.), tracheally instilled surfactant (Inst.Surf.), or nebulized saline (Neb. Saline). PaO2, alveolar-arterial O2 gradient (PAO2-PaO2), PaCO2, and peak inspiratory pressure (PIP) values all significantly improved during 180 min of continuous aerosolization for Neb.Surf. animals compared with pretreatment values (p < 0.01) and with the other two treatment groups (p < 0.01). Although PaO2 and (PAO2-PaO2) values improved for the Inst.Surf. group by 180 min after treatment (p < 0.05), PaCO2 and PIP values were significantly increased 30 min after surfactant instillation (p < 0.05). Neb. Saline animals had no significant changes in physiologic parameters over 180 min. Approximately 8% of the total aerosolized surfactant deposited in lung tissue was recovered from the more severely damaged RML and LING, compared with approximately 50% of the total instilled surfactant recovered from these lobes. This resulted in significantly greater percentages of the total aerosolized surfactant deposited in each of the remaining lobes compared with the percent deposition of instilled surfactant (p < 0.05). Both the underlying pattern of lung injury and the exogenous surfactant delivery technique may influence clinical responses to surfactant therapy in patients with the adult respiratory distress syndrome (ARDS). PMID- 8239153 TI - Role of tidal volume, FRC, and end-inspiratory volume in the development of pulmonary edema following mechanical ventilation. AB - Mechanical ventilation with high peak inspiratory pressure and large tidal volume (VT) produces permeability pulmonary edema. Whether it is mean or peak inspiratory pressure (i.e., mean or end-inspiratory volume) that is the major determinant of ventilation-induced lung injury is unsettled. Rats were ventilated with increasing tidal volumes starting from different degrees of FRC that were set by increasing end-expiratory pressure during positive-pressure ventilation. Pulmonary edema was assessed by the measurement of extravascular lung water content. The importance of permeability alterations was evaluated by measurement of dry lung weight and determination of albumin distribution space. Pulmonary edema with permeability alterations occurred regardless of the value of positive end-expiratory pressure (PEEP), provided the increase in VT was large enough. Similarly, edema occurred even during normal VT ventilation provided the increase in PEEP was large enough. Furthermore, moderate increases in VT or PEEP that were innocuous when applied alone, produced edema when combined. The effect of PEEP was not the consequence of raised airway pressure but of the increase in FRC since similar observations were made in animals ventilated with negative inspiratory pressure. However, although permeability alterations were similar, edema was less marked in animals ventilated with PEEP than in those ventilated with zero end-expiratory pressure (ZEEP) with the same end-inspiratory pressure. This "beneficial" effect of PEEP was probably the consequence of hemodynamic alterations. Indeed, infusion of dopamine to correct the drop in systemic arterial pressure that occurred during PEEP ventilation resulted in a significant increase in pulmonary edema. In conclusion, rather than VT or FRC value, the end inspiratory volume is probably the main determinant of ventilation-induced edema. Hemodynamic status plays an important role in modulating the amount of edema during lung overinflation but does not fundamentally modify the characteristics of this edema which is consistently associated with major permeability alterations. These results may be relevant for ventilatory strategies during acute respiratory failure. PMID- 8239154 TI - Net uptake of lactate by rabbit hindlimb during hypoxia. AB - Lactate uptake by normoxic tissues is a pH-dependent phenomenon that occurs with increases in arterial lactate concentration. In this study we sought to determine the effect of hypoxia on lactate uptake by the rabbit hindlimb at different arterial blood pH (pHa). Two groups of rabbits were subjected to 20 min of arterial hypoxemia (PaO2 approximately 20 mm Hg). One group was allowed to maintain normal pHa (approximately 7.40 to 7.45; n = 5), whereas the other group was hyperventilated, resulting in hypocarbia and alkalemia (pHa > 7.50; n = 5). Lactate uptake was computed from measurements of femoral blood flow and the arterio-venous lactate difference. We found decreases in hindlimb O2 consumption during hypoxemia from 0.79 +/- 0.08 to 0.33 +/- 0.06 ml/min in the normal pH group (p < 0.01) and from 0.49 +/- 0.06 to 0.24 +/- 0.02 ml/min in the high pH group (p < 0.01). At that time there was net uptake of lactate by the hindlimb of the normal pH group (8.1 +/- 2.0 mumol/min; p < 0.001) and lactate release by the alkalemic group (1.61 +/- 1.0 mumol/min). Furthermore, hindlimb lactate uptake was associated with increased glucose consumption (p < 0.001). We hypothesize that the mechanisms responsible for lactate uptake by resting skeletal muscle during hypoxia may be similar to those that regulate lactate uptake during normoxic exercise. PMID- 8239155 TI - Endogenous nitric oxide in single exhalations and the change during exercise. AB - The concentrations of endogenous nitric oxide (NO) and CO2 were measured in exhaled air from healthy humans. During a single exhalation, NO concentrations peaked when the CO2 level reached its plateau. However, whereas CO2 levels remained at a plateau throughout the exhalation, NO concentrations declined after having peaked. After breathholding (5 to 60 s) NO peaked at a higher level than during normal breathing. Nitric oxide concentrations in exhaled air decreased during physical exercise (50 to 100 W). However, taking the increased ventilatory minute volume into account, excretion of NO markedly increased during exercise. The data are compatible with the hypothesis that the NO detected in exhaled air is formed preferentially in the terminal and respiratory bronchioles, and they suggest increased production of NO into exhaled air during exercise and during hyperventilation at rest. PMID- 8239156 TI - Detection and correction of hypoxemia associated with air travel. AB - Hypoxemia occurs during air travel because the pressure in aircraft cabins is not maintained equal to barometric pressure at sea level. Identification of patients who will develop significant arterial hypoxemia is most conveniently achieved by exposing these individuals to inspired oxygen tensions similar to those experienced during air travel. We utilized commercial Venturi devices with nitrogen as a source gas to lower inspired oxygen tensions (114 and 99 mm Hg) to simulate exposure to altitude (6,700 and 10,000 feet) that occurs with air travel. Oxygen was administered by nasal cannulas to correct the induced hypoxemia. Eleven normal subjects had baseline oxygen saturations of 97.3% as determined by ear oximetry. Exposure to reduced oxygen tensions for 20 min using 35 and 40% Venturi devices caused minimal O2 desaturation (94.8 and 93.8%, respectively). Mean baseline arterial oxygen saturation was 93.2% in 11 patients with chronic obstructive lung disease. Oxygen saturation fell to 89.5 and 87.5%, respectively, with exposure to the two levels of reduced oxygen tensions. Hypoxemia was corrected with administration of O2 by nasal cannulas at rates of 1.2 and 1.5 L/min, respectively. We conclude that individual patients can be evaluated conveniently for possible development of hypoxemia associated with air travel using available Venturi devices. This approach also permits determination of the oxygen therapy needed to correct hypoxemia. PMID- 8239157 TI - Small airways dimensions in asthma and in chronic obstructive pulmonary disease. AB - The purpose of this study was to compare the dimensions of the peripheral airways in fatal asthma with those from patients with nonfatal asthma, mild COPD, and normal lung function. Lung specimens from eight individuals who had fatal asthmatic attacks were obtained at postmortem and compared with similar specimens from three asthmatic patients who died of an unrelated cause and four specimens obtained from known asthmatic patients who required lung resection for tumor. These 15 asthmatic lungs were also compared with lungs resected for peripheral neoplasms from 15 patients with normal airway function (FEV1, % of predicted > 85) and 15 patients with mild chronic airflow obstruction (FEV1, % of predicted < 85). All membranous airways with a long-short diameter ratio of 3:1 or less were examined. The smooth muscle and the tissue areas external and internal to the muscle layer were traced using a Bioquant BQ System 4. The same system was used to evaluate the fraction of the submucosa and adventitia taken up by blood vessels. The adventitial, submucosal, and muscle area of the asthmatic airways were greater than those of COPD and control (p < 0.01), and the muscle area was greater in COPD than in control lungs (p < 0.05). These parameters were also greater in the 8 patients with fatal asthma compared with the 7 patients with nonfatal asthma (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239158 TI - Airway inflammation in smokers with nonobstructive and obstructive chronic bronchitis. AB - To assess the manifestation and location of airway inflammation in smokers with chronic bronchitis (CB) or chronic obstructive pulmonary disease (COPD), we lavaged the airways of 12 smokers with CB and 11 smokers with COPD and coexisting CB (OCB). For comparison, the airways of 5 asymptomatic smokers (AS) and 10 healthy nonsmokers (HNS) were lavaged. In all cases, the first lavage aliquot, labeled "bronchial lavage" (BL), was processed separately from the four subsequent aliquots, which were combined and labeled "bronchoalveolar lavage" (BAL). The composition of BL and BAL fluids indicate an ongoing inflammatory process in the airways of all three groups of smokers. CB patients with obstruction had significantly lower concentrations of inflammatory cells in the BL and BAL fluids compared with subjects with nonobstructed CB. Furthermore, airway obstruction, indicated by a reduced FEV1, was significantly correlated with the concentrations of glutathione (p < 0.001), myeloperoxidase (MPO; p < 0.01), and eosinophil cationic protein (ECP; p < 0.01) in BAL fluids. Taken together, these findings suggest that the manifestations of inflammation present in the airways of smokers with CB are different in those who have developed obstruction compared with those who have not. PMID- 8239159 TI - Effects of inhaled capsaicin in heart-lung transplant patients and asthmatic subjects. AB - We have studied the effects on FEV1 of inhaled capsaicin in concentrations of 3 x 10(-6) to 3 x 10(-1) mg/ml and methacholine (1 to 16 mg/ml) in 15 heart-lung transplant (HLT) patients who had undergone recent transbronchial lung biopsy to determine the relationships in chronically denervated lungs between these different forms of airway hyperreactivity and inflammation. A total of 10 normal subjects and 17 asthmatic subjects were included for comparison. Capsaicin caused bronchodilation in eight HLT patients (FEV1 rising by 6.4 to 26.8%) and bronchoconstriction in two (fall in FEV1 of 7.2 and 7.6%). By contrast, seven asthmatic subjects developed bronchoconstriction after capsaicin (fall in FEV1 5.6 to 40.4%); the remaining 10 asthmatic subjects showed no response. Bronchial hyperresponsiveness to methacholine was most evident in the asthmatic subjects, but six HLT patients demonstrated a > or = to 20% fall in FEV1 with < or = 8 mg/ml of methacholine. All normal subjects were nonresponsive to both agents, and all normal and asthmatic subjects, unlike HLT patients, coughed with capsaicin. No relationship existed between the methacholine and capsaicin responses. In the HLT patients neither form of airway responsiveness was related to the degree of inflammation seen on transbronchial lung biopsy. The results suggest that in normal subjects, although it provokes cough, inhaled capsaicin causes little airway narrowing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239160 TI - Assessment of airway tone in asthma. Comparison between double lung transplant patients and healthy subjects. AB - We investigated the hypothesis that asthmatic patients have an increased cholinergic tone by measuring tracheobronchial cross-sectional areas during transient voluntary apnea. This allowed us to assess bronchomotor tone without the influence of changes in lung recoil or lung volume. Three groups of subjects with potentially different levels of tracheobronchial tone were studied: 14 healthy volunteers (N), 18 stable asthmatic patients (A), and 10 double lung transplant recipients (T). Using the acoustic reflection technique, we measured changes in tracheobronchial cross-sectional areas during short periods (5 to 10 s) of voluntary apnea. In a subset of subjects, studies were repeated before and after the inhalation of the muscarinic antagonist ipratropium. During breath holding, glottis and extrathoracic trachea remained unchanged but intrathoracic tracheal area decreased by 30 +/- 8% (mean +/- standard error of the mean) in N, by 27 +/- 3% in A, and by 9 +/- 4% in the T group. Bronchial areas decreased by 24 +/- 8% in N, by 45 +/- 3% in A, and by 10 +/- 4% in T. These differences among groups were statistically significant at the tracheal and bronchial levels (p < 0.05), and ipratropium significantly inhibited this airway constriction (p < 0.05) only in the asthmatic group. Assuming that changes in cross-sectional airway areas voluntary apnea reflect airway tone, these results support the view that in humans this tone is mainly vagally controlled and that it is significantly increased in asthmatic compared with nonasthmatic subjects. PMID- 8239161 TI - Cessation of long-term treatment with inhaled corticosteroid (budesonide) in children with asthma results in deterioration. The Dutch CNSLD Study Group. AB - Inhaled corticosteroid has been shown to be effective in the management of asthma. However, there is a lack of studies that assess the effect of cessation after long-term treatment with inhaled corticosteroid. This question was addressed in 28 children with stable asthma, aged 11 to 18 yr of age, who had completed 28 to 36 months of treatment with inhaled corticosteroid (budesonide 200 micrograms 3 times/day) and inhaled beta-2-agonist (salbutamol 200 micrograms 3 times/day). The children were randomized in a 1:2 ratio in a double-blind study either to continue budesonide (n = 8) during a period of 6 months or to decrease the dose of budesonide (n = 20) within 2 months, followed by placebo for 4 months. Treatment with salbutamol 600 micrograms daily was continued in both groups. Eight children from the tapering-off group withdrew, mainly due to symptoms of asthma, compared with none in the continuous treatment group. Five patients in the tapering-off group experienced exacerbations for which prednisolone was given, compared with none in the continuous treatment group. After tapering-off, symptoms of asthma and additional bronchodilator use increased, and both FEV1% predicted and PD20 histamine (provocation dose of histamine causing a 20% fall in FEV1) decreased, whereas these all remained unchanged in the group that continued treatment with inhaled corticosteroid. We conclude that in this study long-term treatment with 600 micrograms budesonide daily suppressed underlying mechanisms of asthma, but did not cure the disease. PMID- 8239162 TI - Nitrogen dioxide and respiratory illnesses in infants. AB - Nitrogen dioxide is an oxidant gas that contaminates outdoor air and indoor air in homes with unvented gas appliances. A prospective cohort study was carried out to test the hypothesis that residential exposure to NO2 increases incidence and severity of respiratory illnesses during the first 18 months of life. A cohort of 1,205 healthy infants from homes without smokers was enrolled. The daily occurrence of respiratory symptoms and illnesses was reported by the mothers every 2 wk. Illnesses with wheezing or wet cough were classified as lower respiratory tract. Indoor NO2 concentrations were serially measured with passive samplers place in the subjects' bedrooms. In stratified analyses, illness incidence rates did not consistently increase with exposure to NO2 or stove type. In multivariate analyses that adjusted for potential confounding factors, odds ratios were not significantly elevated for current or lagged NO2 exposures, or stove type. Illness duration, a measure of illness severity, was not associated with NO2 exposure. The findings can be extended to homes with gas stoves in regions of the United States where the outdoor air is not heavily polluted by NO2. PMID- 8239163 TI - Passive smoking and lung function in cystic fibrosis. AB - The relationship between passive exposure to cigarette smoking and objective measures of health was examined in 340 patients with cystic fibrosis attending a large hospital-based clinic. Patients who came from households with smokers did not differ from those living in smoke-free households in terms of nutritional status, clinical scores, spirometry, or colonization with Pseudomonas. The number of cigarettes smoked in the household was not significantly related to nutritional status, clinical score, spirometry, or hospitalization. Similar results were found when children 6 to 11 yr of age were analyzed separately, except that height percentile was negatively related to the number of cigarettes smoked in the household. The effects of household exposure to cigarette smoke were further evaluated by analyzing changes in nutritional status, clinical score, and spirometry over a 15-yr period among patients whose families never, always, stopped, or started smoking during this time. Height percentile increased slightly during this interval among those whose households never smoked, whereas no change occurred among patients whose households always smoked, and a decline was seen among patients whose households quit. These differences were statistically significant. Patients whose households never smoked had consistently higher pulmonary function measurements than did patients whose families always smoked, although the differences were not statistically significant. The rates of decline were similar in these two groups. Patients whose households stopped smoking had significantly lower pulmonary functions at the end of the study than did subjects whose households never smoked.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239164 TI - Benefits of continuous positive airway pressure during exercise in cystic fibrosis and relationship to disease severity. AB - The aim of this study was to determine the benefits of CPAP applied during exercise in patients with cystic fibrosis (CF). A total of 33 CF patients with a wide range of lung function were studied. Pulmonary function tests were measured at rest. Endurance tests (80% of previously determined Wpeak) were performed on a bicycle ergometer with and without CPAP (5 cm H2O). Oxygen saturation (SaO2) was monitored by oximetry. Transdiaphragmatic pressure (Pdi) was measured in 7 patients. We found significant correlations between indices of disease severity (NIH score, FEV1, % of predicted, and RV/TLC) and the effects of CPAP on VO2, Pdi, and dyspnea score. CPAP reduced isotime (defined as the last common minute of exercise) VO2 and dyspnea in those patients with more severe lung disease, but these values tended to increase slightly in the patients with only mild lung disease. The change in dyspnea score related to changes in endurance time and VO2. In many patients isotime SaO2 was improved with CPAP, with the largest changes observed in those patients with severe disease. The decreases in VO2, Pdi, and dyspnea score with CPAP in patients with severe lung disease suggest that CPAP can reduce the work of breathing and increase exercise tolerance in patients with CF. These beneficial effects of CPAP during exercise in CF patients are related to disease severity. PMID- 8239165 TI - Decreased leukocyte deformability after acute cigarette smoking in humans. AB - Acute cigarette smoking increases the sequestration of neutrophils in the lungs of humans. This may be due to the delayed transit of cells in the pulmonary microcirculation, which may result from a reduction in cell deformability as suggested by in vitro studies of smoke-exposed neutrophils. In order to support this hypothesis we wished to determine if a reduction in leukocyte deformability could be measured in whole blood exposed to smoke in vitro or in vivo. Whole blood filterability, which largely reflects leukocyte deformability, was measured as the pressure developed by filtration of diluted whole blood through a micropore membrane. Whole blood filtration pressures did not change when blood was exposed to smoke in vitro or in venous blood after acute smoking in vivo. However, arterial blood sampled from chronic smokers during acute smoking showed a consistent reduction in leukocyte deformability associated with a small increase in plasma elastase. To assess whether these changes were induced by oxidants in cigarette smoke, we measured the levels of the antioxidant glutathione (GSH), erythrocyte (RBC) membrane fragility, and products of lipid peroxidation in plasma and RBC in blood exposed to smoke in vivo and in vitro. No change in RBC lipid peroxidation or membrane fragility could be detected after in vitro smoke exposure, possibly because of the high antioxidant capacity of the RBC. However, reduced blood GSH levels and increased levels of lipid peroxidation products were detected in plasma, reflecting oxidant stress. In contrast, we were unable to detect evidence of an increased oxidant burden in blood after acute smoking in vivo, in either arterial or venous blood samples.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239166 TI - Mast cells modulate acute ozone-induced inflammation of the murine lung. AB - We hypothesized that mast cells modulate lung inflammation that develops after acute ozone (O3) exposure. Two tests were done: (1) genetically mast-cell deficient (WBB6F1-W/Wv, WCB6F1-SI/SId) and bone-marrow-transplanted W/Wv mice were exposed to O3 or filtered air, and the inflammatory responses were compared with those of mast-cell-sufficient congenic mice (WBB6F1-(+)/+, WCB6F1-(+)/+); (2) genetically O3-susceptible C57BL/6J mice were treated pharmacologically with putative mast-cell modulators or vehicle, and the O3-induced inflammatory responses were compared. Mice were exposed to 1.75 ppm O3 or air for 3 h, and lung inflammation was assessed by bronchoalveolar lavage (BAL) 6 and 24 h after exposure. Relative to O3-exposed W/Wv and SI/SId mice, the mean numbers of lavageable polymorphonuclear leukocytes (PMNs) and total BAL protein concentration (a marker of permeability) were significantly greater in the respective O3-exposed normal congenic +/+ mice (p < 0.05). Mast cells were reconstituted in W/Wv mice by transplantation of bone marrow cells from congenic +/+ mice, and O3-induced lung inflammation was assessed in the mast-cell-replete W/Wv mice. After O3 exposure, the changes in lavageable PMNs and total protein of mast-cell-replete W/Wv mice were not different from age-matched normal +/+ control mice, and they were significantly greater than those of sham-transplanted W/Wv mice (p < 0.05). Genetically susceptible C57BL/6J mice were pretreated with a mast-cell stabilizer (nedocromil sodium), secretagogue (compound 48/80), or vehicle, and the mice were exposed to O3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239167 TI - Oropharyngeal flora in asthma and in chronic obstructive pulmonary disease. Indigenous oropharyngeal microorganisms in outpatients with asthma or chronic obstructive pulmonary disease. AB - In vitro and in vivo studies have shown that various strains of "viridans streptococci" (nongroupable alpha-hemolytic streptococci) inhabiting the oropharynx suppress the growth of gram-positive and gram-negative microorganisms. We conducted an inventory of the oropharyngeal flora from ambulatory asthma and chronic obstructive pulmonary disease (COPD) patients and a control group to examine the interaction between viridans streptococci and potential pathogens in vivo. In addition, the difference in colonization patterns of these bacteria was studied. Oral washings from 195 patients, 48 asthma (24.6%), 147 COPD (75.4%), and 157 control subjects were examined microbiologically on two occasions with a 2-wk interval, resulting in a total of 384 and 295 oral washings, respectively. All patients were in a stable phase of disease throughout the study. The distribution of low (< or = 10(4)/ml) or high (> or = 10(5)/ml) concentrations of viridans streptococci did not differ substantially between asthma or COPD patients and control subjects. Potentially pathogenic microorganisms found in a low (< or = 10(4)/ml) or high (> or = 10(5)/ml) concentration were equally distributed between the two groups. Staphylococcus aureus and beta-hemolytic streptococci were found significantly less often in the asthma and COPD group (p < 0.005 and p < 0.0005, respectively), but the prevalence of Enterobacteriaceae species was significantly higher (p < 0.0005). No correlation was found between the concentration of viridans streptococci and the prevalence of gram-negative microorganisms. These findings suggest that viridans streptococci are probably not responsible for growth control of gram-negative microorganisms in vivo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239168 TI - Human immunodeficiency virus production by alveolar lymphocytes is increased during Pneumocystis carinii pneumonia. AB - Several factors are known to upregulate in vitro HIV expression by infected T cells, such as certain cytokines and cell-cell interactions. The effect of such factors, involved in immune responses, has never been evaluated in relation to HIV production by infected cells in vivo. To do so, we assessed HIV production by enriched alveolar and blood lymphocytes using a quantitative p24 antigen coculture. This was performed in 32 HIV-seropositive subjects in relation to their pulmonary infectious status, that is, with no current infection (Group 1, n = 17) or with Pneumocystis carinii pneumonia (PCP) (Group 2, n = 15). We showed that HIV core p24 antigen production by enriched alveolar lymphocytes is strongly related to the presence or absence of PC. Although the prevalence of p24-positive alveolar lymphocytes cocultures was 100% in subjects with PCP (15 of 15) compared with 65% in those without lung infection (p = 0.02), all blood lymphocytes tested were positive. In addition, the viral production of alveolar lymphocytes was 40 fold and 8-fold increased at Days 4 and 6 of the culture period in Group 2 compared with Group 1, respectively (p < 0.005). Finally, p24 antigen production by alveolar lymphocytes was significantly higher than that by the corresponding blood lymphocytes in subjects with PCP (p = 0.03) and lower in subjects without lung infection (p = 0.025). Altogether, these data strongly suggest that the HIV burden is markedly enhanced in the lung during PCP. This increased viral production is, at least initially, compartmentalized to the affected organ. PMID- 8239169 TI - The detection of Aspergillus spp. by the polymerase chain reaction and its evaluation in bronchoalveolar lavage fluid. AB - Invasive pulmonary aspergillosis (IPA) is an important cause of mortality and morbidity in the immunocompromised host. However, the diagnosis of this condition may be difficult, and it is sometimes missed because of the lack of sensitivity of available tests. Therefore, we used polymerase chain reaction (PCR)-based amplification of fragments of genes-encoding alkaline proteases from Aspergillus fumigatus and A. flavus to detect these organisms in bronchoalveolar lavage fluid specimens. The predicted size of the product (747 base pairs) after amplification of A. fumigatus was larger than that for A. flavus (690 base pairs). The reaction was highly sensitive (after amplification of 500 fg of A. fumigatus DNA, product could be detected by Southern analysis), and it was specific for A. fumigatus and A. flavus. Bronchoalveolar lavage fluid from four immunosuppressed patients with proved or probable IPA was positive by this assay (sensitivity, 100%); in addition, the sample from one patient with possible IPA was PCR-positive. Only one specimen from 18 immunosuppressed patients with no evidence of IPA was PCR positive (specificity, 94.4%). Five of 28 bronchoalveolar lavage samples from nonimmunosuppressed patients were PCR-positive, probably representing colonization of the respiratory tract. PCR-based detection may prove useful in the diagnosis of IPA. PMID- 8239170 TI - Effect of free radical scavengers on endotoxin-induced respiratory muscle dysfunction. AB - Recent studies have suggested that free radicals contribute to the diaphragmatic dysfunction observed in sepsis. However, previous work has not determined which species of free radicals are responsible for producing these effects or whether the intercostal muscles are affected similarly during sepsis. The purpose of this study was to examine these issues using a hamster model of endotoxin-mediated sepsis in which diaphragm and intercostal muscle function was assessed on muscle strips excised from these animals after killing. Several groups of animals were studied, including animals injected with (1) saline, (2) endotoxin, (3) endotoxin plus active PEG-SOD, a superoxide scavenger, (4) endotoxin plus active PEG catalase, a hydrogen peroxide scavenger, (5) endotoxin plus DMSO, a hydroxyl scavenger, and (6) endotoxin plus denatured PEG-SOD. We found that endotoxin administration elicited significant reductions in diaphragm and intercostal muscle contractility. In each of the three groups of animals to which active free radical scavengers were administered, the effects of endotoxin were attenuated. Denatured PEG-SOD did not protect the respiratory muscles from endotoxin-mediated dysfunction, however. These data indicate that both the diaphragm and intercostal muscles are affected similarly by sepsis; moreover, several free radical species (superoxide ions, hydrogen peroxide, and hydroxyl ions) play a role in mediating this type of injury. PMID- 8239171 TI - Alveolar macrophage surface carbohydrate expression is altered in interstitial lung disease as determined by lectin-binding profiles. AB - Cell-surface-associated glycoconjugates play important roles in cellular functions such as antigen presentation and cell adhesion, functions that may be modulated in patients with interstitial lung disease. Because carbohydrate residues can be recognized by specific lectins, we designed our study to establish baseline data for bronchoalveolar-lavage-derived cells from normal volunteers and to compare the lectin-binding properties of these cells with cells recovered from patients presenting with interstitial lung disease. Cells were obtained from patients with idiopathic pulmonary fibrosis (n = 10), patients with sarcoidosis (n = 20), and patients receiving amiodarone without evidence of clinical lung disease (n = 10) as well as from normal volunteers (n = 8). To determine the pattern of cell-surface glycoconjugate expression on alveolar macrophages (AM), we used a panel of 21 fluorochrome-coupled plant lectins and employed flow cytometry to determine their binding to AM. The labeling profiles of AM were found to be highly reproducible for normal subjects. At the lectin concentrations used for this study, some lectins showed very little binding to AM and some displayed intermediate binding, but the majority of the lectins labeled nearly all AM in samples. Fluorescence intensity varied characteristically for cells labeled with different lectins, providing further refinement and permitting discrimination beyond that provided by data restricted to percent of labeling. AM from patients with interstitial lung disease showed increased binding for the plant-derived lectins PNA, UEA-I, BSL-I, VVL, and SJA compared with AM from normal subjects, being most augmented for AM from patients with idiopathic pulmonary fibrosis. Because peripheral blood monocytes from normal subjects show a higher percentage of labeling with PNA, UEA-I, SJA, and BSL-I than did AM, the increased expression of binding sites for these four lectins by AM from patients with interstitial lung disease may reflect the influx of immature blood monocytes and/or the emergence of a proinflammatory macrophage phenotype. This study demonstrated heterogeneous expression of surface carbohydrate residues by AM and blood monocytes from normal subjects and alterations in carbohydrate receptor expression in interstitial lung disease. Lectin-binding properties may prove useful, therefore, in the evaluation of mononuclear phagocyte populations in interstitial lung disease, especially by the identification of functional subsets and/or changed activation states. PMID- 8239172 TI - Endogenous opioid system during inspiratory loading in patients with type I diabetes. AB - To investigate the activity of the endogenous opioid system in patients with insulin-dependent diabetes mellitus during ventilatory stress situations, we measured plasma beta-endorphin levels in six male and five female diabetic patients breathing against fatiguing inspiratory resistive loads. The patients had to generate with each inspiration an esophageal pressure (Pes) 80% of maximum until they were exhausted and could no longer develop target Pes. The loaded breathing run was repeated three times with a 1-min interval between each run. Duty cycle, tidal volume, and breathing frequency were constant in all tasks. For each run plasma beta-endorphin levels were measured, inspiratory effort sensation assessed using a modified Borg scale, and inspiratory muscle endurance evaluated by the length of time the task could be maintained (Tlim). A group of 11 sex-, age-, height-, and weight-matched healthy individuals served as control subjects. Tlim was significantly lower in the diabetic patients. Evaluating respiratory effort during the three test runs in control subjects at a time corresponding to the break point of loaded breathing in patients showed significantly lower Borg ratings in the control group than in the patient group. Baseline plasma beta endorphin was significantly lower in the diabetic patients (10.6 +/- 2.1 versus 27.0 +/- 4.2 pg/ml, p < 0.01). Additionally, whereas resistive loaded breathing caused a further increase in plasma beta-endorphin concentration in the control group (p < 0.005), absolutely no increase was found in the diabetic patients. We conclude that the endogenous opioid system in insulin-dependent diabetic patients does not respond to stress caused by breathing against fatiguing inspiratory resistive loads. PMID- 8239173 TI - Effect of posture and breathing route on genioglossal electromyogram activity in normal subjects and in patients with the sleep apnea/hypopnea syndrome. AB - Patients with the sleep apnea/hypopnea syndrome (SAHS) often have more apneas supine that sitting. We have shown radiologically that although the retropalatal airway narrows on lying down, the retroglossal airway widens. We have thus investigated the effect of posture on genioglossal EMG activity in 10 normal subjects and 10 patients with SAHS (58 +/- 29 SD apneas + hypopneas/h) using peroral intramuscular EMG electrodes. Data were analyzed by three-way analysis of variance, with diagnosis, posture, and route as factors. Peak inspiratory and tonic expiratory genioglossal EMG were both than sitting, with no significant difference between normal subjects and SAHS higher (p < 0.001) supine patients, although there was a trend (p < 0.09) toward the supine posture having a greater effect on peak inspiratory EMG in the SAHS patients. There was no significant effect of breathing route on either peak inspiratory (p > 0.9) or tonic expiratory (p > 0.8) genioglossal EMG, but there were significant differences between the groups (p < 0.01), the SAHS patients having higher and the normal subjects lower EMG tone with nasal in comparison with oral breathing on both inspiration and expiration. This study therefore shows that both body posture and breathing route are important determinants of genioglossal EMG tone. PMID- 8239174 TI - Chronic upper airway obstruction produces right but not left ventricular hypertrophy in rats. AB - With airway obstruction there is a decrease in inspiratory intrathoracic pressure. This could lead to increased venous return to the right ventricle (RV) and increased afterload imposed on the left ventricle (LV). Chronic upper airway obstruction, caused by either upper airway lesions or obstructive sleep apnea, is a cause of congestive heart failure because of a chronic resistive load imposed on the respiratory system. To determine the effects of chronic upper airway obstruction on RV and LV in adolescent rats, we chronically obstructed the trachea so as to considerably increased inspiratory esophageal pressure excursion (-3.7 +/- 2.2 to -29.4 +/- 10.1 cm H2O). Rats were studied at 7 wk (Group 1) and at 1 yr (Group 2) after tracheal banding. Sham-operated time-matched rats served as controls. In neither group was there evidence of arterial hypoxemia, but in both groups there was chronic hypercapnia (PCO2, approximately 51 mm Hg; bicarbonate, 27 to 28 mEq/ml). Hemoglobin was also normal in both groups, confirming the absence of chronic hypoxia. There were no significant differences between obstructed and control rats in lung, liver, and LV weight to body weight ratio. However, RV weight to body weight ratio was increased in obstructed rats compared with that in control rats in both groups by approximately 50% (p < 0.005). Thus, chronic normoxic airway obstruction leads to evidence of RV but not LV hypertrophy. We conclude that the mechanical effects of airway obstruction impose a chronically increased afterload on the RV, probably caused by venous return effects, but they have relatively little effect on the LV. PMID- 8239175 TI - Exertional breathlessness in patients with chronic airflow limitation. The role of lung hyperinflation. AB - There is considerable intersubject variability in the perceived intensity of breathlessness for a given level of activity among patients with chronic airflow limitation (CAL). To examine possible factors contributing to this variability we compared breathing pattern parameters, dynamic operational lung volumes, and Borg dyspnea ratings in 23 patients with severe CAL and in 10 healthy age-matched normal subjects during cycle ergometry to symptom-limitation. Patients with CAL had significantly (p < 0.01) higher levels of ventilation (% maximal voluntary ventilation) for a given work rate (slope of VE(%MVV)/WR(% pred max) = 1.51 +/- 0.18 versus 0.63 +/- 0.10; mean +/- SEM) and greater dynamic lung hyperinflation (DH) (change [delta] in end-expiratory lung volume [EELVdyn] = +0.31 +/- 0.11 L versus -0.16 +/- 0.22 L). Compared with normal subjects at a standardized VE (30 L/min), the CAL group was more breathless Borg = 4 +/- 1 versus 2 +/- 1, p < 0.01) and hyperinflated (EELVdyn = 75 +/- 3 versus 46 +/- 6% TLC, p < 0.001; end inspiratory lung volume [EILVdyn] = 85 +/- 3 versus 67 +/- 5% TLC, p < 0.01). Within the CAL group, change in Borg ratings correlated with delta VE(%MVV) (r = 0.77, p < 0.001) and with slope of VE(%MVV)/WR(% pred max) (r = 0.48, p < 0.01). Regression analysis selected delta EILVdyn (or delta inspiratory reserve volume [delta IRVdyn]) from various dynamic ventilatory parameters as the strongest predictor of delta Borg (r = 0.63, p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239176 TI - Morphometric analysis of airways in Sjogren's syndrome. AB - Sjogren's syndrome is characterized by a decrease in exocrine gland secretion and the destruction of secretory tissue and cells, that is, salivary and lacrimal glands. Although patients with Sjogren's syndrome have chronic cough and sometimes sputum, we have little knowledge concerning the morphology of their airways. We performed morphometric analysis of goblet cells and bronchial glands and compared them with intraluminal mucus volume by measuring the mucus-occupying ratio (MOR) in the airways of patients with Sjogren's syndrome. A total of 6 nonsmoking patients with Sjogren's syndrome who had no significant prolonged airway infections (Group SG) (6 women, 55 +/- 2 yr) were compared with four control patients with no pulmonary diseases (Group NL) (1 woman and 3 men, 64 +/- 2 yr) and five patients with chronic bronchitis (Group CB) (1 woman and 4 men, 52 +/- 5 yr). Autopsied lungs from 15 patients were used for morphometry by a digitalizing computer. Area proportion of bronchial glands (gland %) to bronchial wall, goblet cell-occupying ratio to total epithelial layer (goblet %), and MOR were measured in central (3-8 mm in diameter) and peripheral airways (2 mm or less in diameter). Gland % and goblet % in central and MOR in peripheral airways from Group SG were significantly larger than those from Group NL: 15 +/- 2 versus 6 +/- 1% (p < 0.01), 6 +/- 1 versus 2 +/- 1% (p < 0.05), and 9 +/- 2 versus 0.6 +/- 0.3% (p < 0.05), respectively. Further, these values from Group SG did not significantly differ from those from Group CB.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239177 TI - Ozone-induced airway inflammation in human subjects as determined by airway lavage and biopsy. AB - Ozone (O3) is a major constituent of urban air pollution. The acute effects of the inhalation of O3 at ambient or near-ambient concentrations on bronchoalveolar lavage (BAL) end points consistent with a distal lung inflammatory response have been well documented in human subjects. Animal toxicologic studies have shown that the airway is also a major site of O3-induced injury and inflammation. To date, no studies have confirmed this finding in human subjects. Effects of O3 on the proximal airways are not adequately studied by BAL, which is primarily influenced by events occurring in the terminal bronchioles and alveoli. We hypothesized that O3 causes injury and inflammation in the airways in addition to that previously documented to occur in the distal lung. We performed isolated lavage of the left mainstem bronchus and forceps biopsy of the bronchial mucosa in a group of 14 healthy, athletic subjects 18 h after exposure to 0.20 ppm O3 for 4 h during moderate exercise in order to assess this possibility. We followed an identical protocol in a similar group of 12 subjects exposed to filtered air. The mean (SD) total cell count and the lactate dehydrogenase (LDH) concentration in the isolated airway lavage were significantly greater after O3 than after air, 13.9 (20.5) versus 4.9 (5.4) cells/ml x 10(4) and 18.9 (11.2) versus 9.6 (9.0) U/L, respectively. Morphometry (2,070 neutrophils/cm2 of tissue for O3 and 330 neutrophils/cm2 of tissue for air) demonstrated that O3 exposure induced an acute inflammatory cell influx into the airway.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239178 TI - Recurrence of sarcoidosis in pulmonary allograft recipients. AB - Lung transplantation is a potentially curative therapy for the end-stage pulmonary sequelae of sarcoidosis. We reviewed the course of five lung allograft recipients with underlying sarcoidosis (S) at the University of Pittsburgh Medical Center and compared them with a control group (C) of 44 contemporaneous transplant recipients with other respiratory diseases. Sarcoid granulomata have developed in the allografts of 4 S, although these lesions have not yet been demonstrated to result in clinically significant abnormalities. In comparison with C, sarcoidosis patients had significantly greater mean grades of acute rejection during the first 3 months after transplantation (2.1 +/- 0.3 versus 1.6 +/- 0.1, S and C, respectively, p < 0.042) and larger proportions of lung biopsies showing more than mild acute rejection (40 versus 18%, p < 0.012) and lymphocytic bronchitis (30 versus 13%, p = 0.02), as well as a greater percentage of polymorphonuclear leukocytes in BAL returns (34.9 +/- 5.4 versus 19.0 +/- 1.6, p < 0.01). The two groups did not differ, however, in frequency of obliterative bronchiolitis, survival, or pulmonary function. We conclude that lung transplant recipients with underlying sarcoidosis are very likely to develop recurrent disease in the allograft and have more severe acute rejection responses, especially in the first weeks after transplantation. Pulmonary transplantation appears to be an efficacious therapy for end-stage sarcoidosis, but the long-term sequelae of the increased acute rejection and recurrent sarcoidosis in the allograft remain to be determined. PMID- 8239179 TI - Interleukin-1 receptor antagonist expression in sarcoidosis. AB - Sarcoidosis is a systemic granulomatous disease with a marked propensity for involvement of the pulmonary parenchyma and thoracic lymphatic system. This granulomatous process is characterized by aggregations of mononuclear cells, multinucleated giant cells, and variable degrees of fibrosis. The agent(s) responsible for the initiation of the inflammatory granulomatous process remain unknown. Interleukin-1 beta (IL-1) is a cytokine that has been shown to possess potent proinflammatory properties and is likely to play a role in mediating many of the immunopathologic events observed in sarcoidosis. Despite the degree of granulomatous inflammation, both the pulmonary and systemic pathogenic changes associated with sarcoidosis have a remarkable propensity for spontaneous resolution. The interleukin-1 receptor antagonist (IRAP), an endogenous inhibitor of IL-1 bioactivity, may have a critical role as an in vivo immunomodulator of IL 1-dependent granulomatous inflammation of sarcoidosis. In this study we demonstrate constitutive expression of IRAP mRNA and antigen from bronchoalveolar lavage fluid cells and cell-free fluid, respectively, obtained from both normal subjects and patients with sarcoidosis. However, immunolocalization of IRAP was found to be significantly localized to the sarcoid granuloma as compared with the uninvolved lung interstitium. Our findings indicate that IRAP expression is compartmentalized (granuloma) within the interstitium of patients with sarcoidosis. Thus, IRAP may function as an important in vivo immunomodulator of granulomatous inflammation. PMID- 8239180 TI - Dynamic upper airway imaging during awake respiration in normal subjects and patients with sleep disordered breathing. AB - The effects of respiration on upper airway caliber were studied using cine computed tomography (CT) in 15 normal subjects, 14 snorer/mildly apneic subjects, and 13 patients with obstructive sleep apnea. All subjects were scanned in the supine position during awake nasal breathing. Eight-millimeter-thick axial slices were obtained at four anatomic levels from the nasopharynx to the retroglossal region every 0.4 s during a respiratory cycle. Tidal volume measured from an integrated pneumotachograph signal was correlated with slice acquisition during inspiration and expiration to generate loops comparing upper airway area and tidal volume. In all three subject groups and at all anatomic levels studied, there were significant dimensional changes in upper airway caliber during the respiratory cycle. The major findings in this investigation include: (1) the upper airway was significantly smaller in apneic than normal subjects, especially at the retropalatal low and retroglossal anatomic levels; in apneic patients the airway had an anterior-posterior configuration unlike the normal airway, which had a horizontal configuration with the major axis in the lateral direction; (2) in all three subject groups, little airway narrowing occurred in inspiration, suggesting that the action of the upper airway dilator muscles balanced the effects of negative intraluminal pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239181 TI - Giant cell interstitial pneumonitis. Disease recurrence in the transplanted lung. AB - Recurrence of the original disease in the transplanted organ is well reported in renal transplant recipients. There have been no previously fully documented cases of recurrence of the original disease after lung transplantation. We report a patient who underwent single-lung transplant in 1990 for end-stage respiratory failure secondary to biopsy-proved giant cell interstitial pneumonitis (GIP). There was no further industrial exposure. Surveillance bronchoscopies and biopsies post-transplant demonstrated eosinophils and giant cells in the bronchoalveolar lavage of both lungs, and in biopsies of the transplanted organ. Two years after successful transplantation the patient deteriorated and underwent open lung biopsy, which demonstrated not only bronchiolitis obliterans but also the classic features of GIP. There was no evidence of inorganic particles in the transplanted lung. Autopsy confirmed the presence of numerous giant cells characteristic of GIP with associated fibrosis throughout the transplanted lung. Although tungsten and other inorganic particles were again demonstrated in the native lung, there was no evidence of tungsten particles in the transplanted lungs. We believe that this case documents recurrence of the original disease after lung transplantation. The absence of unusual inorganic particles in the transplanted lung in the face of the classic picture of GIP is highly suggestive of an autoimmune mechanism for this occupation-associated disease. The appropriateness of transplant in the management of this lung disease should be reviewed further. PMID- 8239182 TI - Mycobacterium kansasii presenting as an endobronchial lesion. AB - Four patients underwent bronchoscopy to evaluate a cavitary lesion (one patient), pulmonic infiltrate (two patients), and a lingular mass (one patient). In all cases, an endobronchial obstruction was found. In three cases, forceps biopsy specimens revealed acid-fast bacilli, and all four cultures subsequently yielded only Mycobacterium kansasii. Three of the four patients had AIDS. Endobronchial obstruction has been reported for Mycobacterium tuberculosis and other atypical mycobacteria; however, we believe this to be the first reported series of M. kansasii presenting as an endobronchial obstruction. Unlike the endobronchial lesions seen with M. tuberculosis and other atypical Mycobacterium, the lesions seen with M. kansasii responded favorably to therapy. PMID- 8239183 TI - Liver transplantation and pulmonary gas exchanges in hypoxemic children. AB - Hypoxemia in cirrhotic patients is well documented. One of the possible causes of this association seems to be the presence of functional intrapulmonary shunts. The extent of the ventilation/perfusion ratio (VA/Q) abnormalities and their regression after orthotopic liver transplantation has been previously studied in adults by the multiple inert gas elimination technique. We report here a similar study in three children where the hypoxemia was the main indication for early liver grafting, although the liver function was still preserved at that time. Their hypoxemia was almost exclusively caused by a right to left shunt (VA/Q = 0) with a minimal amount of poorly ventilated but well perfused areas (Low VA/Q). This association may explain the poor response of the arterial oxygen pressure to an increased inspired oxygen concentration. Despite these very large VA/Q mismatches, the children underwent successful liver transplantations, resulting in a regression of the intrapulmonary shunt, as demonstrated by multiple inert gas elimination technique, and compatible with a normal life. PMID- 8239184 TI - The effect of forceps size on the adequacy of specimens obtained by transbronchial biopsy. AB - This study prospectively compared the diagnostic yield of transbronchial biopsies using large and small forceps (cup sizes, 3 x 2 x 0.9 versus 2 x 1.5 x 0.6 mm, respectively). Diagnostic yield was compared by a pathologist, blinded to the size of forceps used on the basis of the relative amount of tissue obtained, alveolar tissue obtained, and ability to ascertain a histopathologic diagnosis. Large forceps obtained significantly more tissue than did small forceps (20 of 27 patients [74%] versus five of 27 patients [19%], p < 0.005, with similar amounts obtained in two patients). Also, large forceps obtained significantly more alveolar tissue than did small forceps (16 of 22 patients [73%] versus six of 22 patients [27%], p < 0.05, with no alveolar tissue obtained in five patients). In 18 of the 27 patients, biopsies performed resulted in nonspecific diagnoses, including fibrosis or chronic inflammation. All nine of the patients with a specific diagnosis were ultimately proved to have sarcoidosis. There was a trend toward more of these patients having noncaseating granulomas obtained with the large forceps than with the small forceps (seven of nine patients versus four of nine patients). No difference was observed in the amount of postbiopsy bleeding with either forceps. We conclude that large forceps used for transbronchial biopsy yield more tissue and more alveolar tissue than do small forceps. These findings may have an impact on the diagnostic yield in some diseases such as sarcoidosis. PMID- 8239185 TI - Investigation and management of pulmonary hypertension in chronic obstructive pulmonary disease. PMID- 8239186 TI - Guidelines for the initial management of adults with community-acquired pneumonia: diagnosis, assessment of severity, and initial antimicrobial therapy. American Thoracic Society. Medical Section of the American Lung Association. PMID- 8239187 TI - Effect of smoking on the alveolar macrophage phenotype. PMID- 8239188 TI - A study of 101 patients treated with extraperitoneal endoscopic laparoscopic herniorrhaphy. AB - One hundred twenty-two hernias were repaired in 101 male patients through a total extraperitoneal approach. Patients ranged from 18 to 78 years old. All repairs were done with polypropylene mesh. Five patients (5%) required conversion to an open or transabdominal approach. Patients have been followed from 6 to 20 months, with a mean of 12 months. No recurrence has developed to date. Complications included urinary retention, groin hematoma, trocar site infection, and lateral femoral cutaneous nerve neuralgia. Six patients underwent simultaneous extraperitoneal endoscopic pelvic lymph node dissections, and two patients had varicoceles repaired simultaneously. Patients returned to usual activity within 1 week. PMID- 8239189 TI - Obturator hernia: a report of eight cases. AB - Obturator hernia is a rare problem that occurs predominantly in elderly, emaciated women. The diagnosis of obturator hernia is difficult to establish in most patients before an operation is performed. Both delayed diagnosis and delayed surgical intervention contribute to high mortality. From 1978 to 1992, eight cases of obturator hernia involved elderly females who had received operations at this hospital. Resection of the incarcerated bowel was required. The often debilitated condition of these patients and the frequent delay of diagnosis combined to produce significant operative mortality. PMID- 8239190 TI - Laparoscopic gallbladder delivery aided with nasal speculum. PMID- 8239191 TI - Rare presentation of anastomotic iliac artery false aneurysm: rupture with formation of ilio-iliac arteriovenous fistula. AB - A rare complication of aortoiliac bypass procedure resulting in the formation of an acute ilio-iliac arteriovenous fistula was presented. The latter was a first manifestation of anastomotic iliac pseudoaneurysm. This is only the second such case reported in the English literature. Iliac anastomotic pseudoaneurysm although rarely detected, is a potentially lethal complication that has to be repaired close to the time of its detection. PMID- 8239192 TI - Mature fibrous cyst formation after Marlex mesh ventral herniorrhaphy: a newly described pathologic entity. AB - Two patients developed seromas overlying Marlex mesh repairs of ventral hernias. After repeated unsuccessful attempts at aspiration of the fluid, each patient underwent formal exploration and excision of the cystic mass to the level of the mesh. Histology showed thick, fibrous, trabeculated cysts. Postoperatively, one patient had prolonged closed suction drainage. In the other, suction drains had scant output, but she developed a recurrence of serous fluid, which resolved with two aspirations. CONCLUSIONS: 1) Mature fibrous cyst is a rare complication of Marlex mesh herniorrhaphy; 2) Chronic seroma collections may be etiologic; 3) Surgical excision and closed suction drainage, rather than simple aspiration, are required for treatment; 4) Prolonged postoperative drainage may be needed to prevent recurrence. PMID- 8239193 TI - Hepatic arterioportal fistula. AB - Hepatic arterioportal fistula (HAPF) is uncommon, arising largely from either a ruptured hepatic artery aneurysm or from penetrating trauma. It is being encountered increasingly, secondary to rising numbers of percutaneous transhepatic procedures. We will describe five cases of HAPF seen at Emory University Hospital over the last 5 years and conclude with a review of the current status of treatment of this condition. HAPF occurred secondary to ruptured hepatic artery aneurysm in two patients, operative injury in two patients, and after trauma in one patient. The predominant clinical manifestation in these patients were complications of portal hypertension, most commonly gastrointestinal bleeding. Angiography remains the definitive diagnostic procedure and it was used to accurately diagnose HAPF in all cases. However, duplex ultrasonography is rapidly becoming a useful screening tool and was the initial imaging modality in the most recent case. Several factors determine the therapeutic approach including the location of the HAPF and associated comorbidity. Due to the development of portal hypertension even asymptomatic fistulas should be treated. In general, surgery is the procedure of choice for extrahepatic fistula, while embolization is optimal for intrahepatic fistulas. Embolization is also useful for patients in whom associated comorbidity prohibits surgical treatment. The goal of surgery is to interrupt the fistula, to maintain hepatic arterial flow, and to close the defect in the portal vein. PMID- 8239194 TI - The role of infection in outcome of Multiple Organ Failure. AB - It is widely assumed that infections are the principal cause and primary outcome determinant of the syndrome of Multiple Organ Failure (MOF) in critically ill patients. Infections are frequent in these patients, but the prevention and treatment of infections may not influence the course of MOF. This study tested the hypothesis that infections play a decisive role in the outcome of MOF. Data were gathered concurrently on all adult patients admitted over an 18-month period to a non-cardiac surgical ICU at a university hospital and recorded in a computer database. Sepsis was defined as a state characterized by at least three of the following: fever, tachycardia, leukocytosis or leukopenia, increased cardiac index, reduced systemic vascular resistance, and hypercatabolism manifested by nitrogen-wasting. The presence of an infection was not required for the diagnosis of sepsis. Mild sepsis was defined as the presence of three or four parameters. Severe sepsis was defined as the presence of five or six parameters. MOF was defined as the development of dysfunction of at least two of the following major organ systems: cardiac, gut, pulmonary, renal, cerebral, and hepatic. Of 749 admissions, 73 patients developed MOF. Thirty four (47%) had a documented source of infection, 37 (51%) had positive blood cultures, and all had sepsis. Hospital mortality was 66 percent (48 of 73 patients). Death could not be predicted by bacteremia (P > 0.25), nor by the presence of an infectious source (P = 1.0), but was strongly associated with severe sepsis (P < 0.0005).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239195 TI - Bilateral breast cancer. AB - Seventy-eight confirmed cases of second primary breast cancer in the contralateral breast were encountered over a 22-year period in 1332 women with invasive breast cancer treated in our department. Tumors were grouped into those simultaneously detected in both breasts or within 6 months of each other (synchronous, 1.6%) and those detected within more than 6 months (metachronous, 4.2%). The mean interval between metachronous cancers was 117 months. Patients with bilateral tumors were more likely to have a family history of breast cancer than those with unilateral disease. Women with metachronous tumors tended to be younger when diagnosed with the first carcinoma as compared with those having unilateral or synchronous bilateral cancers. No differences were noticed in size and lymph node status between the first or second tumor of bilateral cases in comparison to patients with unilateral disease. Significantly more (P < 0.05) first metachronous tumors were found to be lobular invasive cancers. Histopathologic type of the first tumor was the same as the second in 62.8 per cent of all cases. Concordance of estrogen receptor status between bilateral tumors was 71.4 per cent. Our results indicate that the risk of developing, a contralateral breast cancer is related to the patient's age, family history of breast cancer, and lobular histology of the tumor. PMID- 8239196 TI - The limited role of total parenteral nutrition in the management of pancreatic pseudocyst. AB - Total parenteral nutrition (TPN) for the nonoperative treatment of acute pancreatic pseudocyst has been of hypothetical benefit. We reviewed pseudocyst hospital admissions in 40 patients treated with TPN who had serial imaging studies. The mean cyst size was 7.4 cm on presentation, decreasing to 5.6 cm after nonoperative treatment with TPN (mean 32.5 days). After a nonoperative period, 68 per cent of cysts regressed, completely in 14 per cent, partially in 54 per cent. Except for a patient with cyst-related obstructive jaundice, there were no complicated pseudocysts. Only 12 (28%) patients underwent cyst drainage. Fifteen patients (35%) sustained catheter-related complication, which included sepsis (26%), pneumothorax (9%), hydropneumothorax (2%), and septic right atrial thrombosis (2%), in the course of hospitalization. The majority of TPN-treated patients had a clinical and radiographic regression of their pseudocyst. However, the increased risk of catheter-related complications in this group suggests that this therapy should be limited to patients who are unable to sustain enteral nutrition. PMID- 8239197 TI - Revascularized diabetic limbs: positional changes in regional perfusion index. AB - We measured pre- and postoperative ankle:brachial index (ABI), regional perfusion index (RPI = foot/chest transcutaneous oxygen tension [TcpO2]), and variation in RPI with limb elevation in 22 ischemic lower extremities of 20 patients to compare ABI and RPI measurements for quantifying limb perfusion and analyze perioperative positional changes in RPI. Measurements were compared, using t tests, with all limbs grouped according to severity of clinical ischemia and, again, according to presence or absence of diabetes. Preoperative mean and mean post-revascularization increases in ABI values ranged from 0.27 to 0.48 and 0.40 to 0.54, respectively; corresponding RPI values ranged from 0.18 to 0.45 and 0.48 to 0.60, respectively. Pre- and postoperative decreases in RPI with elevation ranged from 0.07 to 0.11 and 0.11 to 0.23, respectively. ABI and RPI values were equally effective in assessing clinical ischemia preoperatively and increased perfusion postoperatively, regardless of degree of ischemia or diabetes. Upon elevation, all limbs exhibited larger decrements in blood flow to the skin postoperatively compared to preoperatively, as estimated by RPI. However, postoperative positional decrease in RPI was greater in diabetics compared with nondiabetics (0.23 +/- 0.12 vs 0.12 +/- 0.06; P < 0.05), suggesting postoperative elevation of diabetic limbs with ischemic skin lesions may be unadvisable. PMID- 8239198 TI - Penetrating injuries to the neck: a safe, selective approach to management. AB - The most appropriate management of patients with penetrating injuries to the neck remains a controversial issue. In order to determine the safety and efficacy of a selective approach to the management of such patients, a retrospective review of our experience with penetrating injuries to the neck at Northwestern Memorial Hospital over the past 5 years since the designation as a Level I trauma center was undertaken. A total of 30 patients fulfilled entry criteria. Twelve patients underwent immediate operative exploration based upon clinical indications present at admission. Seventeen patients underwent further diagnostic evaluation, including angiography in 17 and contrast esophagography in eight. Endoscopy was used infrequently. The mortality rate was 13.3 per cent, there were two negative cervical explorations, and there were no missed injuries. The results support the application of a selective approach to the operative management of penetrating injuries to the neck. PMID- 8239199 TI - Laboratory parameters as predictors of operative outcome after major abdominal surgery in AIDS- and HIV-infected patients. AB - Surgeons throughout the country are frequently asked to consult on acquired immunodeficiency syndrome (AIDS) and human immunodeficiency virus (HIV)-infected patients experiencing abdominal pain. Disease processes vary remarkably within this population and often occur with confusing presentations and unusual pathologies related to the immunocompromised state. With the increased awareness and treatment of HIV infection, it can be anticipated that many patients will require surgery for secondary complications of AIDS, in addition to surgical problems unrelated to HIV infection. Twenty-five patients diagnosed with HIV infection underwent major abdominal surgery between 1986 and 1990 at The Mount Sinai Medical Center. Those patients classified as having AIDS had a longer post procedure hospitalization (19 days vs 9 days; P < 0.05) and a higher mortality rate (33% vs 10%). All of the patients who underwent appendectomy survived with few complications. Excluding appendectomy patients, operative mortality was predicted by low serum albumin (P < 0.001). In addition, preoperative hematocrits were considerably lower in non-survivors. Total serum protein and total WBC counts were not predictors of operative outcome. PMID- 8239200 TI - Oncotic pressure, albumin and ileus: the effect of albumin replacement on postoperative ileus. AB - The effect of decreased colloid oncotic pressure, as seen in hypoalbuminemia and hypoproteinemia, upon intestinal function has been well delineated in the surgical literature. Patients undergoing abdominal aortic aneurysm resection or aortoiliac or aortofemoral bypass grafts are almost uniformly hypoalbuminemic postoperatively; with these two facts in mind, a prospective, randomized clinical study was undertaken to identify the role of serum albumin concentration on the length of postoperative ileus in this population. The main hypothesis was that patients whose albumin levels dropped below 3.5 gm/dL would have a more prolonged postoperative hospital course as a result of delay in return of bowel function when compared with those patients in whom the low albumin levels were exogenously acutely replenished to > 3.5 gm/dL. Albumin was replaced to a level greater-than or equal to 3.5 g/dL in one group of 37 patients (AR), with a control group of 32 patients (NR) not receiving any albumin. Return of bowel function was measured by the postoperative day that flatus was documented, as well as the postoperative day oral intake was resumed. Mean values were determined for each group, and t tests did not reveal a significant difference in postoperative day of flatus (AR mean = 4.06 days, NR mean = 4.16 days) or postoperative day of oral intake (AR mean = 4.0, NR mean = 3.75). Additional comparisons between the groups involving the number of postoperative days until a regular diet was begun (AR mean = 6.06, NR mean = 5.48) and length of postoperative hospital stay (AR mean = 9.16, NR mean = 8.43) failed to reveal significant differences.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239201 TI - Fatal cerebral gunshot wounds: factors influencing organ donation. AB - Patients with clinical brain death following head injury are important potential cadaveric organ donors. We analyzed our series of cranial gunshot wounds with particular attention to the frequency and patterns of organ donation after fatal injuries. Sixty-six patients with gunshot wounds to the head, including 59 with intracranial involvement (43 male, average age 26 years) were seen during a 4 year period. Injuries were limited to the head in 50 of 59 patients. Overall mortality was 66 per cent. Predictors of mortality included Glasgow Coma Scale (GCS) of six or less (93%), self-inflicted gunshot wounds (75%), and computed tomography (CT) findings of bihemispheric injury (87%) or ventricular injury (82%). Of the 39 patients who died, 28 met standard criteria for brain death, and nine of these went on to organ procurement. Thirteen families refused donation, and six patients were not harvested for reasons including old age, pregnancy, suspicion of AIDS, coroner refusal, and failure to pursue consent. Principles essential to maximal organ retrieval include: 1) Recognition that patients suffering cerebral gunshot wounds represent potential organ donors and that certain factors are predictive of mortality; 2) Critical care/trauma team approach with standardized management and timely declaration of brain death; 3) Early search for family members and prompt notification of organ procurement agencies; 4) Sensitivity to cultural issues influencing donation; and 5) Programs to increase public awareness of organ donation. PMID- 8239202 TI - Peripheral fat necrosis after penetrating pancreatic trauma: a case report. AB - Peripheral fat necrosis (PFN), a rare complication of pancreatitis, has been reported previously in association with blunt pancreatic trauma. A patient who developed peripheral fat necrosis after penetrating pancreatic trauma and needed bilateral above-the-knee amputations to treat complications of lower extremity fat necrosis is reported. PMID- 8239203 TI - [Estimation of Neisseria meningitidis carriers among healthy subjects in contact with patients with meningococcal sepsis in the Madrid area]. AB - We have studied 238 asymptomatic school children which had come in contact with one of 5 cases of meningococcal sepsis, which occurred between January 1992 to May 1992. The study was performed at the corresponding schools and oropharynx samples were immediately put into Thayer-Martin agar plates. The cases were identified as serotype C in three occasions and type B in two cases. The total carrier prevalence was 1.68%, all of which were serotype B. We did not find a significant relationship between asymptomatic carriers and meningococcal sepsis cases. In addition, all contacts were treated with rifampin. Control cultures were made after 15 days of the treatment and 100% of the carriers had negative cultures. PMID- 8239204 TI - [Sudden death syndrome in infancy. I. Methods for the identification of high risk infants]. AB - Sudden infant death syndrome (SIDS) occurs with a high frequency, about 2 in every 1,000 live newborns, and could be prevented by home monitoring. However, identification of infants at high risk must first be made. In this paper, we report the epidemiological factors of 36 selected infants with a well known high risk of SIDS. Two different scorings for the evaluation of high risk SIDS were compared and used in these children. The Sheffield scoring showed high risk in 11 (30.5%) of the cases versus 7 (9.7%) in the normal control group (p = 0.013). By using the multistage scoring method, 27 (75.0%) of the cases had a high risk versus 8 (11.1%) in the controls (p < 0.001). Nevertheless, by the latter method, too many normal infants (41 = 56.9%) were predicted to have a moderate risk of SIDS. The multistage test seems more useful than the Sheffield test, but has the limitation of having to be evaluated at one month of age, whereas results of the Sheffield test are obtained at birth. The epidemiological circumstances could be different in every social level or geographical area, so the usefulness of each scoring procedure must be evaluated in the normal population previous to being used in study groups. PMID- 8239205 TI - [Invasive Haemophilus influenzae type B infections in infancy (1981-1990)]. AB - We report the epidemiology of invasive Haemophilus influenzae type B infections. The study is based on 58 children and was carried out during the years from 1981 to 1990. Sixty-three percent of the patients had meningitis, 10.3% had arthritis, 10.3% had epiglottitis, 8.6% had cellulitis and 6.9% had pneumonia. Thirteen percent of the cases were children who were less than 6 months old, 34.4% were less than 12 months old, 70.6% were less than 24 months of age and 93% were less than 4 years old. Among the latter, 90% of the children with epiglottis or pneumonia were 24 months or older compared with 16% of the patients with meningitis, arthritis or cellulitis (p < 0.001). Sixty-three percent of the isolated strains of Haemophilus influenzae type B were resistant to ampicillin and 19.2% of the strains were resistant to ampicillin and chloramphenicol. PMID- 8239206 TI - [Bronchial hyperreactivity in infancy]. AB - Airway responsiveness may be defined as the case with which airways narrow upon exposure to physical and chemical stimuli and drugs. Bronchial hyperresponsiveness reflects an excessive bronchoconstricting response to these stimuli. Although in part it may be genetically mediated, a number of environmental factors (allergens, viruses and contaminants) are also involved in increasing bronchial responsiveness through airway inflammation. We review the concept, etiology, pathogeny, clinical expression, diagnosis, preventative measures and treatment of bronchial hyperresponsiveness in pediatrics. PMID- 8239207 TI - [Children of heroin-addicted parents: a risk group. Study of 110 cases]. AB - In order to know the medical and social problems of children of heroin addicted parents (one or both), one-hundred and nineteen patients were studied. All children had been hospitalized suffering from different diseases. The age of these children ranged from 1 day to 17.3 years (19.6 +/- 30.9 months). Seventy four of the children were less than 1 year old. The total number of admissions was 159 (range 1-14), representing 2.575 days of hospitalization. The most frequent diagnoses were infectious diseases (gastrointestinal and respiratory), deficiency related diseases (malnutrition and anemia) and parental neglect. Maltreatment was suspected in 44 children (physical: 10, severely neglected: 26, abandonment: 8). Eight children died during the study (AIDS: 4, maltreatment: 2, other causes: 2). Nine parents also died (4 from heroin overdose). The social and family environment of these children, with an increased risk for delinquency and marginality, as well as frequent hospitalizations, make these children a special group at risk for social and health problems which demands multidisciplinary attention and understanding. PMID- 8239208 TI - [Balloon (Rashkind) atrio-septostomy in transposition of great vessels in the neonatal period]. AB - Analysis of the balloon atrial septostomy (BA) for treatment of the transposition of the great arteries (TVG) during the neonatal period will allow the evaluation of some of the changes that have occurred in the diagnosis and therapy of this congenital cardiopathy. We performed a retrospective survey of the management, evolution complications of 24 BA as a palliative technique in TGV cases, admitted to the Neonatal Intensive Care Unit during a 5 1/2 year period. The aim of the study was to evaluate the factors that indicated a bad prognosis by comparing patients that are still alive (21) with those that are dead (3). The mean age at BA performance was 4.5 +/- 5.5 days. The biological constants prior to catheterization were normal, maintaining good oxygenation, administering prostaglandins in 80% of the patients and mechanical ventilation in 25% of the patients. BA outcome was considered as good in 68%, moderate in 12% and bad in 20% of the cases. Intra-catheterization complications, mainly hemorrhages and bradycardia, were suffered by 40% of the patients and one patient died during catheterization. We could see a progressive arterial desaturation due to the lack of efficacy of the BA. As bad post-catheterization outcome factors were found: hypotension, arrhythmia, hemorrhage, greater need of dobutamine, volume expansion and mechanical ventilation. Our principal conclusion is that, although BA has improved the prognosis of newborns with TGV, because of the progressive worsening of the patients during the months following the BA, it is necessary to perform early corrective surgery. PMID- 8239209 TI - [Perioperative management of systemic pulmonary shunts in the neonatal period]. AB - We have analyzed 36 newborns (19 males and 17 females), with cyanotic cardiopathies in whom a systemic-pulmonary shunt had been performed. These patients were admitted to the Neonatal Intensive Care Unite between January 1985 and June 1990. We studied the perioperative events with the aim of describing the general features of this palliative surgery in the neonatal period and to determine the factors which indicate a bad prognosis. The age at admission was 10.5 +/- 16.5 days and at surgery was 19.5 +/- 20 days. Cardiopathy types included: 13 pulmonary atresias, 9 pulmonary stenoses, 6 D-transposition of the great arteries and 8 tetralogies of Fallot. Of these patients, 83% required prostaglandin administration before surgery. Cardiac catheterization was performed in 54% (in 1/3 balloon atrioseptostomy was performed). The mean diameter of the pulmonary branches was 3.5 +/- 0.7 mm. In 89%, a modified Blalock Taussig shunt and in 11% a central aorto-pulmonary shunt were performed. The size of the prosthetic graft used was 4 mm in diameter in 77% of the cases, 5 mm in 20% and 3 mm in 3% of the patients. The mortality rate was 27% (intraoperative = 0%, early postoperative = 16%, late postoperative = 11%) with two critical periods: the initial 48 postoperative hours and the reoperation. Bad prognosis factors related to mortality are; preoperative (shorter gestational age and low newborn weight), operative (performance of a central aortopulmonary shunt, surgical ligation of the ductus during operation) and postoperative (arrhythmias, metabolic acidosis in the immediate postoperative period, lower oxygen saturation at 24 hours after the operation, hemorrhage). Our data are compared with other reports of systemic-pulmonary shunt in the neonatal period. PMID- 8239210 TI - [Determining factors of nutritional status in a pediatric surgical unit]. AB - Nutritional assessment was made in 134 pediatric patients before surgery in order to analyze the influence of age, sex anthropometric nutritional index and type of surgical operation (urgent vs elective). The anthropometrical index, hematology measurements and serum protein levels were compared by using a one-way ANOVA and a linear correlation was performed when appropriate. The type of surgery and age of the patient were the factors which influence nutritional status the most among the pre-operatively screened surgical pediatric patients. In addition, pre albumin, retinol-binding protein and acute-phase proteins have been shown to be affected by stress and appear to be more sensitive indicators of nutritional state than comparison of the types of surgery. PMID- 8239211 TI - [Persistent negative tuberculin test in a case of urinary tract tuberculosis in childhood]. PMID- 8239212 TI - [Neonatal Salmonella enteritidis infection]. PMID- 8239213 TI - [Familial homozygotic infantile hypercholesterolemia treated with lipoprotein apheresis during a year]. PMID- 8239214 TI - [Fraser syndrome, renal agenesis and fetal ascites]. PMID- 8239215 TI - [Congenital benign cystic mesothelioma]. PMID- 8239216 TI - [Intradural anesthesia in a newborn infant with congenital heart defect. Alternative to general anesthesia]. PMID- 8239217 TI - [Lacto-bezoar in a full-term neonate]. PMID- 8239218 TI - [Sigmoid volvulus as first manifestation of Hirschsprung disease in a school-age child]. PMID- 8239219 TI - [Unusual presentation of Pneumocystis carinii infection]. PMID- 8239220 TI - [Post-traumatic splenic pseudocyst]. PMID- 8239221 TI - [Advances in the knowledge of osteochondrodysplasias: new international classification and prenatal diagnosis]. PMID- 8239222 TI - [Prevalence of Neisseria meningitidis carriers in a school population in the metropolitan area of Madrid]. AB - The prevalence of healthy carriers of Neisseriae meningitidis has been addressed by studying oropharynx samples of 1,172 schoolchildren in Madrid. The sample design took into consideration social status, age and sex. The total prevalence of healthy carriers was 2.30%, with 66% or these in the six year old age group (RR = 2.88, p < 0.005). Serotype B was found in 74% of the carriers, again mainly in the six year old age group, and 18.5% were serotype A, with most of them being found in the fourteen year age group. Serotype A carriers were females in 80% of the cases (p = 0.04). Serotype C was only found in six year old males. The prevalence of healthy carriers of Neisseriae meningitidis in our school-age population, in a non-epidemic situation, is in accord with results reported from other countries around us and agrees with previous reports of a higher prevalence among younger children and in males. In addition, the association between sex and some serotypes is confirmed. PMID- 8239223 TI - The effect of inhibition of 5-lipoxygenase by zileuton in mild-to-moderate asthma. AB - OBJECTIVE: To evaluate the effectiveness of inhibiting the formation of the 5 lipoxygenase products of arachidonic acid by the 5-lipoxygenase inhibitor zileuton in the treatment of mild-to-moderate asthma. DESIGN: Randomized, double blind, placebo-controlled study. SETTING: University hospitals and private allergy and pulmonary practices. PATIENTS: A total of 139 persons with asthma who had a forced expiratory volume in 1 second (FEV1) of 40% to 75% of the predicted value and who were not being treated with inhaled or oral steroids. INTERVENTION: Zileuton, 2.4 g/d or 1.6 g/d, or placebo for 4 weeks. MEASUREMENTS: Airway function, beta-agonist use, and symptoms; inhibition of 5-lipoxygenase assessed by measurement of urinary leukotriene E4 (LTE4). RESULTS: Zileuton produced a 0.35-L (95% CI, 0.25 to 0.45 L) increase in the FEV1 within 1 hour of administration (P < 0.001 compared with placebo), equivalent to a 14.6% increase from baseline. After 4 weeks of zileuton therapy, airway function and symptoms improved, with the greatest improvements occurring in the 2.4 g/d group: This group's FEV1 increased by 0.32 L (CI, 0.16 to 0.48 L), a 13.4% increase, compared with a 0.05-L (CI, -0.10 to 0.20 L) increase in patients taking placebo (P = 0.02). Symptoms and frequency of beta-agonist use also decreased with zileuton, 2.4 g/d. The mean urinary LTE4 level decreased by 39.2 pg/mg creatinine (CI, 18.1 to 60.4 pg/mg creatinine) and 26.5 pg/mg creatinine (CI, 6.6 to 46.5 pg/mg creatinine) in the 2.4 g/d and 1.6 g/d groups, respectively, compared with a slight increase in the placebo group (P = 0.007 and P = 0.05). No difference was noted in the number of adverse events among treatment groups. CONCLUSIONS: Inhibition of 5-lipoxygenase can improve airway function and decrease symptoms and medication use in patients with asthma, suggesting that this inhibition can be useful therapy for asthma. Also, 5-lipoxygenase products may mediate part of the baseline airway obstruction in patients with mild-to-moderate asthma. PMID- 8239224 TI - Hydroxychloroquine compared with placebo in rheumatoid arthritis. A randomized controlled trial. AB - OBJECTIVE: To assess the efficacy of hydroxychloroquine, 400 mg daily, for rheumatoid arthritis. DESIGN: Six-month, double-blind, randomized trial. SETTING: Ambulatory referral clinic in a Mexico City, Mexico, teaching hospital. PATIENTS: A total of 126 patients with early rheumatoid arthritis were randomly assigned to receive hydroxychloroquine, 400 mg/d, or placebo; 121 patients completed the study. RESULTS: Hydroxychloroquine showed a clinically and statistically significant improvement over placebo in joint score (20% greater mean improvement; 10% more patients improved by > 50%); pain (40% greater mean improvement; 19% more patients improved by > 50%); grip strength (22% greater mean improvement; 21% more patients improved by > 50%); patient global assessment (16% more patients stated they had improved); and physician global assessment (12% more patients were judged to have improved). Side effects were mild, and no patients in the hydroxychloroquine group required discontinuation of therapy. Patient compliance with the study medication was high. CONCLUSION: Hydroxychloroquine is moderately effective in early rheumatoid arthritis. PMID- 8239225 TI - An outbreak of gram-negative bacteremia traced to contaminated O-rings in reprocessed dialyzers. AB - OBJECTIVE: To investigate an outbreak of gram-negative bacteremia in an outpatient hemodialysis unit and to identify the source of contaminating bacteria and the route by which bacteria gained access to the bloodstream. DESIGN: A matched-pair, case-control study and a bacteriologic investigation of the hemodialysis unit and the implicated dialyzers. SETTING: A university outpatient hemodialysis unit. PATIENTS: Eleven patients receiving long-term hemodialysis who had a total of 12 episodes of primary gram-negative bacteremia and 12 matched controls. MEASUREMENTS: Clinical and demographic data were obtained for patients and controls. Dialysis unit procedures were observed for compliance with aseptic technique. Cultures of potential environmental sources of bacteria were obtained. Hemodialyzers from bacteremic and nonbacteremic patients were dismantled, and the component parts were cultured. Inoculation of O-rings (from Hemoflow F-80 dialyzer) with bacteria and simulated dialysis were done. RESULTS: During January to October 1988, 12 episodes of primary gram-negative bacteremia caused by Pseudomonas cepacia, Xanthomonas maltophilia, Citrobacter freundii, Acinetobacter calcoaceticus var. anitratus, or Enterobacter cloacae occurred in 11 patients. In 11 episodes, symptoms developed within 3 hours of starting hemodialysis. Intravenous antibiotics were administered for 11 episodes, 3 episodes resulted in hospitalization, and all patients recovered. Case patients were more likely to have received high-flux dialysis with Hemoflow F-80 dialyzers (odds ratio congruent to 11) than were controls. O-rings from dialyzers used by bacteremic patients were culture positive for the organism responsible for bacteremia. Three of the four dialyzers were disinfected using the standard automated method and were recultured 72 hours later; the O-rings of all three dialyzers remained culture positive. Simulated dialysis using dialyzers with contaminated O-rings caused blood pathway contamination despite intervening reprocessing. When the disinfection method for F-80 dialyzers included removal and complete disinfection of the O-rings, O-ring and blood pathway cultures were consistently negative. After this procedure was made routine, no episodes of primary gram-negative bacteremia occurred during the next 6 months. CONCLUSIONS: Because dialyzers with removable headers and O-rings are widely used in patients receiving long-term hemodialysis, disinfection procedures should include measures to ensure adequate disinfection of O-rings. PMID- 8239226 TI - Occurrence of antibodies to Borrelia burgdorferi in patients with nonspirochetal subacute bacterial endocarditis. AB - OBJECTIVE: To determine the prevalence and specificity of antibodies to Borrelia burgdorferi in patients with nonspirochetal subacute bacterial endocarditis and assess whether increased levels of antibodies to B. burgdorferi were attributable to rheumatoid factor. DESIGN: Retrospective case-control study. SETTING: Urban referral center in an area devoid of infected ticks as a source of endocarditis sera. PATIENTS: Sera from 30 consecutive patients with culture-proven subacute endocarditis between 1979 and 1981 were compared with 30 control sera collected between 1989 and 1990. In addition, sera from 20 consecutive patients with rheumatoid arthritis who were positive for rheumatoid factor were collected between 1991 and 1992. Sera were compared with a convenience sample from 15 patients who met the criteria for Lyme disease. MEASUREMENTS: Antibodies to B. burgdorferi were assessed by enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. IgM rheumatoid factor was quantified using solid-phase radioimmunoassay or latex agglutination techniques. RESULTS: Thirteen of 30 patients with endocarditis (43%) compared with 3 of 30 normal controls (10%) had increased levels of antibodies to B. burgdorferi (P < 0.01). Of these 13 patients, only 1 had an immunoblot consistent with previous infection. The others had nonspecific immunoblots: 5 showed isolated 60-kd reactivity; 1 patient had isolated 41-kd reactivity; and 6 had no bands of reactivity. Immunoblots of the 3 controls with increased antibodies showed only isolated 41-kd reactivity. Thus, the specificity of the B. burgdorferi antibody test in patients with endocarditis was only 60% (95% CI, 42% to 78%), compared with 90% (CI, 79% to 100%) in controls. No correlation was noted between IgM rheumatoid factor and antibodies to B. burgdorferi in patients with endocarditis (r = 0.2; P > 0.2). Only 1 of 20 patients with rheumatoid arthritis without known bacterial infections had antibodies to B. burgdorferi. CONCLUSIONS: Although a positive ELISA test for B. burgdorferi may be a "true positive," a positive serologic test alone does not ensure that the clinical problem is due to Lyme borreliosis. Cross-reactive antibodies to shared epitopes between B. burgdorferi and the endocarditis organism may account for the high false-positive results. PMID- 8239227 TI - Coronary revascularization after myocardial infarction in the very elderly: outcomes and long-term follow-up. AB - OBJECTIVE: To determine the outcome of very elderly patients who had coronary revascularization during hospitalization for an acute myocardial infarction. DESIGN: Retrospective cohort study. SETTING: Community-based tertiary-care teaching hospital. PATIENTS: A total of 1215 consecutive patients 80 years and older were hospitalized with a myocardial infarction between 1985 and 1990. The study sample included all 93 patients (8%) who had cardiac catheterization before discharge and had not been excluded from study because of the following: severe valvular disease, absence of significant coronary disease, or death before a decision about revascularization could be made. MEASUREMENTS: Survival, quality of life, and functional status at least 1 year after discharge. RESULTS: After catheterization, 41 patients had angioplasty, 18 had coronary artery bypass surgery, and 34 did not have revascularization. Among the patients alive at discharge, those who had revascularization had a high likelihood of achieving a good or excellent quality of life (angioplasty, 86% [31 of 36]; surgery, 89% [16 of 18]; medical therapy, 44% [11 of 25]) and of being able to care for themselves (angioplasty, 89% [32 of 36], surgery, 89% [16 of 18], medical therapy, 52% [13 of 25]). Mortality rates at 1 year were 24% (95% CI, 15% to 47%) for the angioplasty group, 6% (CI, 0% to 27%) for the surgery group, and 44% (CI, 27% to 62%) for the medical therapy group. In a Cox proportional hazards model that adjusted for clinical, demographic, hemodynamic, and anatomic differences between the groups, the performance of coronary revascularization was associated with increased survival (hazard ratio, 0.42; CI, 0.18 to 0.98). CONCLUSIONS: A small percentage of very elderly patients with complicated acute myocardial infarctions, selected by their physicians for invasive cardiovascular procedures, can tolerate these procedures, avoid serious complications, return to independent living, and have excellent probability of survival. Although our results suggest that coronary revascularization may have benefited these patients, the study design did not permit definite conclusions, and future studies are needed to resolve this important question. PMID- 8239228 TI - Interferon-alpha 2b: a new treatment for polycythemia vera. PMID- 8239229 TI - Primary central nervous system lymphoma. AB - OBJECTIVE: To compare the pathogenesis, clinical presentation, therapy, and prognosis of primary central nervous system lymphoma in immunocompetent persons with these characteristics of the disease in patients with AIDS. DATA SOURCES AND EXTRACTION: All English-language papers published between 1980 and 1992 dealing with either lymphoma and the central nervous system or AIDS were reviewed. Patient characteristics, clinical presentation, histologic findings, and treatment and survival data were extracted from each case report and review. DATA SYNTHESIS: Data were available on 792 patients (from 40 reported series) with non AIDS-associated primary central nervous system lymphoma and 315 patients (from 32 series) with AIDS-associated primary central nervous system lymphoma. Patients with AIDS initially consulted a physician more often when they had global neurologic symptoms compared with patients without AIDS, with more than 50% of the lesions on computed tomographic (CT) scans in patients with AIDS being ring enhancing and multifocal, a pattern rarely described in immunocompetent patients. The overall survival of the patients without AIDS was 18.9 months compared with 2.6 months for patients with AIDS, with substantial differences remaining even for subgroups of patients similarly treated with radiation and chemotherapy. CONCLUSION: Primary central nervous system lymphoma is probably a substantially different disease in persons with and without AIDS with regard to patient characteristics, clinical and radiographic presentation, and prognosis. Recent advances in the treatment of this disease in patients without AIDS have not largely affected patients with AIDS. Substantial improvements in survival in these patients await advances in controlling their human immunodeficiency virus associated disease. PMID- 8239230 TI - Efficacy and cost of low-molecular-weight heparin compared with standard heparin for the prevention of deep vein thrombosis after total hip arthroplasty. AB - PURPOSE: To compare the efficacy, safety, and cost-effectiveness of low-molecular weight heparin with standard heparin for the prevention of deep vein thrombosis after total hip arthroplasty. DATA IDENTIFICATION: Studies were identified by MEDLINE search and review of bibliographies of retrieved articles. Hospital resources used in treating deep vein thrombosis and bleeding complications after total hip arthroplasty were estimated using retrospectively collected data from 447 patients who participated in a recently completed randomized controlled deep vein thrombosis prophylaxis trial at our center. STUDY SELECTION: Randomized controlled trials directly comparing a low-molecular-weight heparin preparation with standard heparin for the prevention of deep vein thrombosis after total hip arthroplasty were potentially eligible for the meta-analysis. DATA EXTRACTION: Data from eligible studies were extracted independently by two of the authors. Multiple regression analysis of data from the patient cohort was used to estimate the effect of deep vein thrombosis and bleeding on length of hospital stay. A hypothetical North American price for low-molecular-weight heparin was determined based on the ratio between low-molecular-weight heparin and standard heparin in France. Costs were based on weighted per-diem hospital expenditures and physician fees for procedures and reported in 1992 U.S. dollars. RESULTS OF DATA SYNTHESIS: Meta-analysis of six eligible trials determined that low-molecular-weight heparin was significantly more effective than standard heparin at preventing deep vein thrombosis after total hip arthroplasty (common odds ratio, 0.72; 95% CI, 0.53 to 0.95). However, this benefit was restricted to the prevention of proximal deep vein thrombosis (common odds ratio, 0.40; CI, 0.28 to 0.59). No significant differences were found in the rates of distal deep vein thrombosis or total, major, or minor bleeding between the two groups. Based on a 2.6 to 1 price ratio between low-molecular-weight heparin and standard heparin, use of low-molecular weight heparin would save the health care system about $50,000 per 1000 patients treated. Sensitivity analysis shows that if the low-molecular-weight heparin/standard heparin price ratio exceeds 3.7 (the threshold value lies between 0.8 and 5.5 based on the extremes of the 95% CI of the common odds ratios for deep vein thrombosis and bleeding complications), use of low-molecular-weight heparin is more expensive. At a price ratio of 10, it would cost more than $250,000 to treat 1000 patients with low-molecular-weight heparin compared with standard heparin or about $5000 for each additional deep vein thrombosis prevented with low-molecular-weight heparin. CONCLUSIONS: Low-molecular-weight heparin is more effective and is at least as safe as standard heparin for the prevention of deep vein thrombosis after total hip arthroplasty. Based on the current French price ratio of low-molecular-weight heparin to standard heparin, the use of low-molecular-weight heparin in North America would result in overall savings in cost; however, the relative cost-effectiveness is critically dependent on the price ratio between the two drugs. Further research is needed to compare the cost-effectiveness of low-molecular-weight heparin with other prophylactic regimens and postoperative deep vein thrombosis management strategies. PMID- 8239231 TI - Syndromes of glucocorticoid resistance. AB - Glucocorticoid resistance results from the partial, albeit apparently generalized, inability of glucocorticoids to exert their effects on target tissues. The condition is associated with compensatory increases in circulating pituitary corticotropin and cortisol, with the former causing excess secretion of both adrenal androgens and adrenal steroid biosynthesis intermediates with salt retaining activity. The manifestations of glucocorticoid resistance vary from chronic fatigue (perhaps a result of glucocorticoid deficiency in the central nervous system) to various degrees of hypertension with or without hypokalemic alkalosis or hyperandrogenism, or both, caused by increased cortisol and other salt-retaining steroids and adrenal androgens, respectively. In women, hyperandrogenism can result in acne, hirsutism, menstrual irregularities, oligoanovulation, and infertility; in men, it may lead to infertility and in children, to precocious puberty. Different molecular defects, such as point mutations or a microdeletion of the highly conserved glucocorticoid receptor gene, alter the functional characteristics or concentrations of the intracellular receptor and appear to cause glucocorticoid resistance. The extreme variability in the clinical manifestations of glucocorticoid resistance and its mimicry of many common diseases can be explained by the overall degree of glucocorticoid resistance, differing sensitivity of target tissues to mineralocorticoids or androgens or both, and perhaps different biochemical defects of the glucocorticoid receptor, with selective resistance of certain glucocorticoid responses in specific tissues. The various different symptoms of classic glucocorticoid resistance and the theoretical potential of this condition to appear surreptitiously emphasize the importance of the glucocorticoid receptor in the pathogenesis of human disease. PMID- 8239232 TI - Generating more generalists: an agenda of renewal for internal medicine. Federated Council for Internal Medicine. PMID- 8239233 TI - The future of internal medicine. AB - Internal medicine may be in its twilight because it has failed to address the shortage of primary care physicians by training more general internists. Data from several sources indicate that progressively fewer persons are entering general internal medicine as opposed to its subspecialties. The reasons for this decline include adverse experiences in medical school, an unfavorable patient mix, declining incomes, and increasing hassles in caring for patients. A series of reforms, such as improving the teaching in medical school, strengthening divisions of general medicine, and establishing financial incentives, are proposed to reverse this trend. Other actions that must be taken include stopping the proliferation of subspecialty certificates, designating and accrediting primary care tracks, and cutting subspecialty positions. Internal medicine's fate is in its own hands, and the discipline must reorient itself to conform to societal needs. PMID- 8239234 TI - Getting into medical school in the good old days: good for whom? The first Nicholas E. Davies Memorial Lecture. AB - Admission to medical school became selective in the 1920s and by the 1960s became increasingly expensive for applicants. The stories of three applicants, a white man who easily walked into medical school in 1908, a black woman who overcame a double hurdle in the 1930s, and a white man who entered medical school by court order, provide insight into the admissions process and its changes. PMID- 8239235 TI - What is the future of internal medicine? PMID- 8239236 TI - Hydroxychloroquine is safe and probably useful in rheumatoid arthritis. PMID- 8239237 TI - Sarcoidosis, liver transplantation, and cyclosporine. PMID- 8239238 TI - Sarcoidosis, liver transplantation, and cyclosporine. PMID- 8239239 TI - Association of cytomegalovirus infection and penile ulcer. PMID- 8239240 TI - Successful immunosuppressive treatment of high-titer postpartum factor VII inhibitor. PMID- 8239241 TI - When and whom to screen. PMID- 8239242 TI - When and whom to screen. PMID- 8239243 TI - When and whom to screen. PMID- 8239244 TI - When and whom to screen. PMID- 8239245 TI - Does aminophylline benefit adults admitted to the hospital for an acute exacerbation of asthma? AB - OBJECTIVE: To determine the effect of adding intravenous theophylline (administered as aminophylline) to nebulizations of albuterol and intravenous methylprednisolone in adults hospitalized for acute asthma. DESIGN: Randomized, placebo-controlled, double-blind study. SETTING: Inpatient service of a tertiary care, university teaching hospital. PATIENTS: 21 adults (22 to 48 years old)--10 in the aminophylline group and 11 in the placebo group. INTERVENTIONS: Nebulized albuterol, 2.5 or 5.0 mg every 0.5 to 4 hours; intravenous methylprednisolone, 60 mg every 6 hours; and either individualized doses of aminophylline or placebo for 48 hours. MEASUREMENTS: Forced expiratory volume in 1 second (FEV1), the number of "as needed" albuterol nebulizations and total dose, asthma symptom scores, and adverse effects. RESULTS: At admission from the emergency department, the mean +/ SD baseline FEV1 was 49% +/- 19% of the predicted value in the aminophylline group and 43% +/- 13% of the predicted value in the placebo group. The improvement in FEV1 at 3 hours was greater in the aminophylline group (29% +/- 23% compared with 10% +/- 10% in the placebo group; mean difference, 19 percentage points; 95% CI, 3 to 35 percentage points; P = 0.023). At 48 hours, FEV1 was 75% +/- 19% of the predicted value in the aminophylline group and 58% +/ 15% of the predicted value in the placebo group (mean difference, 17 percentage points; CI, 0.2 to 34.8 percentage points; P = 0.048). Aminophylline-treated patients required fewer nebulizations of albuterol (10.3 +/- 3.8 compared with 16.4 +/- 5.3; mean difference, -6.1; CI, -10.3 to -1.8) and less total dosage (34 +/- 16 mg compared with 70 +/- 34 mg; mean difference, -36 mg; CI, -60.6 to -11.3 mg P = 0.02). No statistical differences were observed in asthma symptom scores or frequency of adverse effects. CONCLUSIONS: Individualized doses of intravenous theophylline added to frequent nebulizations of albuterol and intravenous methylprednisolone appear to benefit adults admitted to the hospital with acute asthma and are well tolerated when serum concentrations are maintained in the therapeutic range. PMID- 8239246 TI - Predictors of major toxicity after theophylline overdose. AB - OBJECTIVE: To identify patients at high risk for major toxicity after theophylline intoxication who might benefit from early charcoal hemoperfusion. DESIGN: A 67-month prospective study. SETTING: Massachusetts Poison Control System. PATIENTS: 249 consecutive patients referred after theophylline intoxication (defined by a peak serum theophylline concentration > or = 167 mumol/L [30 mg/L]). INTERVENTIONS: Uniform, protocol-directed management recommendations. MAIN OUTCOME MEASURES: Identification of risk factors for major toxicity. RESULTS: 119 patients (48%) not receiving theophylline therapy had acute intoxication; among those receiving such therapy, 92 (37%) had theophylline intoxication because of chronic overmedication and 38 (15%) had acute intoxication. Major toxicity developed in 62 patients (25%); 13 patients (5%) died. Major toxicity was more common in patients with intoxication due to chronic overmedication than in those with acute intoxication who were not receiving theophylline therapy (49% compared with 10%, risk ratio, 4.85; 95% CI, 2.96 to 7.94), even though the former group had lower peak serum theophylline concentrations (283 mumol/L compared with 777 mumol/L, P = 0.001). Logistic regression analysis identified two major factors associated with the development of major toxicity: 1) peak serum theophylline concentrations in cases of acute intoxication and 2) patient age in cases of chronic overmedication. Receiver operating characteristic curve analysis indicated that major toxicity occurred in patients with a peak serum theophylline concentration of greater than 555 mumol/L (100 mg/L) after acute intoxication and in patients older than 60 years (regardless of peak serum theophylline concentration) after chronic overmedication. CONCLUSIONS: Predictors for major toxicity after theophylline intoxication differ by type of overdose. PMID- 8239247 TI - Infectious morbidity associated with long-term use of venous access devices in patients with cancer. AB - OBJECTIVE: To evaluate infectious morbidity associated with long-term use of venous access devices. DESIGN: Prospective, observational study. SETTING: Comprehensive cancer center at a university hospital. PARTICIPANTS: 1431 consecutive patients with cancer requiring 1630 venous access devices for long term use inserted between 1 June 1987 and 31 May 1989. MEASUREMENTS: Quantitative microbiologic tests to identify device-related bacteremia and fungemia, catheter tunnel infection, pocket infection in implantable port devices, and site infections; number of days the device remained in situ and time until infectious morbidity; vessel or device thrombosis and device breakage. RESULTS: At least one device-related infection occurred with 341 of 788 (43% [95% CI, 39% to 47%]) catheters compared with 57 of 680 (8% [CI, 6% to 10%]) completely implanted ports (P < or = 0.001). Device-related bacteremia or fungemia is the predominant infection occurring with catheters, whereas ports have a more equal distribution of pocket, site, and device-related bacteremia. The predominant organisms isolated in catheter-related bacteremia were gram-negative bacilli (55%) compared with gram-positive cocci (65.5%) in port-related bacteremia. The number of infections per 1000 device days was 2.77 (95% CI, 2.48 to 3.06) for catheters compared with 0.21 (CI, 0.16 to 0.27) for ports (P < or = 0.001). Based on a parametric model of time to first infection, devices lasted longer in patients with solid tumors than in those with hematopoietic tumors. Ports lasted longer than catheters across all patient groups. CONCLUSIONS: The incidence of infections per device-day was 12 times greater with catheters than with ports. Patients with solid tumors were the least likely to have device-related infectious morbidity compared with those with hematologic cancers. The reasons for the difference in infectious complications is uncertain but may be attributable to type of disease, intensity of therapy, frequency with which devices are accessed, or duration of neutropenia. PMID- 8239248 TI - Cataract formation after bone marrow transplantation. AB - OBJECTIVE: To evaluate the incidence, time course, and factors associated with cataract formation in bone marrow transplant recipients. DESIGN: Prospective cohort study. SETTING: University Hospitals, Basel, Switzerland. PATIENTS: 197 patients treated with allogeneic or autologous bone marrow grafts at least 180 days before the start of the study. INTERVENTION: Three regimens for bone marrow transplant were used: 74 patients received single-dose, total-body irradiation (TBI), 90 patients received fractionated TBI, and 33 received chemotherapy alone. RESULTS: Three and one half years after single-dose TBI, 51 of the 74 patients (69%) were alive and cataracts had developed in all of these 51 patients. Cataracts developed in 18 of the 90 (20%) patients treated with fractionated TBI, with an 83% (95% CI, 63% to 100%) risk for lens opacification at 6 years. Cataracts developed in only 1 of the 33 (3%) patients treated with chemotherapy alone. Incidence of cataracts is higher and lens opacification occurs earlier after single-dose TBI than after fractionated TBI (P < 0.01). With Cox regression analysis, the use of irradiation (relative risk, 21.0), the mode of irradiation (relative risk, 7.4), and the use of steroid treatment (relative risk, 2.9) for more than 3 months after bone marrow transplantation increased the risk for cataract formation. In contrast, age, sex, and chronic graft-versus-host disease did not influence the rate of cataract development. The probability of requiring cataract surgery after 6 years was 85% (CI, 75% to 95%) for the patients treated with single-dose TBI and 20% (CI, 0% to 49%) for those prepared with fractionated irradiation. CONCLUSIONS: Patients treated with TBI, regardless of fractionation, are likely to have cataracts within 10 years, and some will need surgical repair. Long-term steroid treatment accelerates cataract formation. Preventive measures, such as lens shielding during TBI, should be considered. PMID- 8239249 TI - HIV-1 seroconversion in patients with and without genital ulcer disease. A prospective study. AB - OBJECTIVE: To determine the relative risk for human immunodeficiency virus (HIV 1) seroconversion in patients with and without genital ulcers caused by chancroid, syphilis, and herpes. DESIGN: A prospective cohort study. SETTING: An inner-city, sexually transmitted disease clinic. PATIENTS: Patients seronegative for HIV-1 with and without genital ulcers who were followed for a minimum of 3 months. INTERVENTIONS: Questionnaire to obtain data on demographics, sexual behavior, and illicit drug use; testing for HIV-1 at entry and at a minimum of 3 months after entry; medical examination for the presence or absence of genital ulcer disease. RESULTS: Overall, 758 heterosexual men with no history of injection drug use completed the study; HIV-1 seroconversion occurred in 10 of 344 (2.9%; 95% CI, 1.4% to 5.3%) men with a genital ulcer and in 4 of 414 (1%; CI, 0.2% to 2.5%) without a genital ulcer (relative risk, 3.0; P = 0.05). In a multiple logistic regression analysis, those men with chancroid and a new sexually transmitted disease during follow-up each had about three times the risk for HIV-1 seroconversion (P < or = 0.04). CONCLUSIONS: In this group of heterosexual men, chancroid and repeated acquisition of sexually transmitted diseases appeared to facilitate the sexual transmission of HIV-1. PMID- 8239250 TI - Sudden cardiac death: epidemiology, transient risk, and intervention assessment. AB - PURPOSE: To integrate information from the various disciplines that contribute to the understanding of the cause and prevention of sudden cardiac death: identification of new approaches from applied clinical epidemiology; identification and control of transient risk factors; and evaluation of the results of interventions. DATA SOURCES: A broad range of research reports and interpretations of data from English-language journal articles and reviews, published primarily between 1970 and 1993. The fields of study included epidemiology, experimental electrophysiology, clinical observations, and interventions. STUDY SELECTION: Continuous literature surveys, done in relation to ongoing clinical and experimental research on sudden cardiac death since 1972. DATA EXTRACTION: Included on the basis of relevance to the topics discussed and with confirmation of data and concepts by more than one investigator when available. DATA SYNTHESIS: Information from several disciplines was integrated by the authors to synthesize new ways to view the problem of sudden cardiac death. Quantitative information was used primarily to derive qualitative statements about new perspectives on sudden cardiac death. CONCLUSIONS: Progress in the prevention of sudden death will require development of new approaches, including epidemiologic techniques to address risk characteristics specific to the problem; characterization of triggering events and identification of specific persons at risk for responding adversely to these events; and methods of evaluating outcomes appropriate to the nature of sudden cardiac death. PMID- 8239251 TI - Glucocorticoid therapy for immune-mediated diseases: basic and clinical correlates. AB - Glucocorticoids are pleiotropic hormones that at pharmacologic doses prevent or suppress inflammation and other immunologically mediated processes. At the molecular level, glucocorticoids form complexes with specific receptors that migrate to the nucleus where they interact with selective regulatory sites within DNA; this results in positive and negative modulation of several genes involved in inflammatory and immune responses. At the cellular level, glucocorticoids inhibit the access of leukocytes to inflammatory sites; interfere with the functions of leukocytes, endothelial cells, and fibroblasts; and suppress the production and the effects of humoral factors involved in the inflammatory response. Clinically, several modes of glucocorticoid administration are used, depending on the disease process, the organ involved, and the extent of involvement. High doses of daily glucocorticoids are usually required in patients with severe diseases involving major organs, whereas alternate-day regimens may be used in patients with less aggressive diseases. Intravenous glucocorticoids (pulse therapy) are frequently used to initiate therapy in patients with rapidly progressive, immunologically mediated diseases. The benefits of glucocorticoid therapy can easily be offset by severe side effects; even with the greatest care, side effects may occur. Moreover, for certain complications (for example, infection diathesis, peptic ulcer, osteoporosis, avascular necrosis, and atherosclerosis), other drug toxicities and pathogenic factors overlap with glucocorticoid effects. Minimizing the incidence and severity of glucocorticoid related side effects requires carefully decreasing the dose; using adjunctive disease-modifying immunosuppressive and anti-inflammatory agents; and taking general preventive measures. PMID- 8239252 TI - The Uniform Clinical Data Set: an evaluation of the proposed national database for Medicare's quality review program. AB - The accuracy, reliability, and validity of the Medicare Peer Review Organization (PRO) review process have all been questioned. Evidence concerning the PRO program's effect on cost and quality of care remains lacking. The Health Care Financing Administration has thus committed itself to reform, and the Uniform Clinical Data Set (UCDS) has been proposed as the national database for Medicare's quality review program. The UCDS is an automated, computerized data set designed to standardize the evaluation of quality. It should allow an objective, consistent, and efficient process for peer review, based on explicit decision criteria and on aggregated information about patterns of care and quality. But is this truly so? We review the existing evidence on the UCDS and compare it with the current PRO reviews of quality, concluding that although the UCDS can potentially improve the accuracy and the reliability of data abstraction and the validity of reviews, this remains to be shown. Preliminary data on the UCDS suggest that work is needed before it can meet appropriate expectations for a national database for quality assessments. We also propose a model for reviews of quality in which we show that reviews of care done in the context of internal quality improvement programs will differ in goals and intensity from reviews of care done at the national level. We suggest that the UCDS has a unique, but limited role--that of national surveillance of practice patterns. Detailed assessments of quality are more appropriately done at local or institutional levels. PMID- 8239253 TI - The miracle of the eye. PMID- 8239254 TI - Hemlock or healer? The mercurial reputation of theophylline. PMID- 8239255 TI - The primary prevention of sudden death in patients with coronary artery disease. PMID- 8239256 TI - Identifying Pneumocystis carinii in pleural effusions. PMID- 8239257 TI - Fibrinogen and cardiovascular risk. PMID- 8239258 TI - Interaction of ofloxacin and warfarin. PMID- 8239259 TI - Angiotensin-converting-enzyme inhibitors and chyluria. PMID- 8239260 TI - Does race predispose to angiotensin-associated angioneurotic edema? PMID- 8239261 TI - Subdividing departments of medicine. PMID- 8239262 TI - Subdividing departments of medicine. PMID- 8239263 TI - Treatment of nongonococcal urethritis. PMID- 8239264 TI - Alzheimer's Disease: Amyloid Precursor Proteins, Signal Transduction, and Neuronal Transplantation. Proceedings of a meeting. Zurich, Switzerland, February 12-14, 1993. PMID- 8239265 TI - Second messengers in neuronal signaling. AB - The past decade has witnessed an enormous increase in our knowledge of the variety and complexity of intracellular signaling events that follow receptor binding on the cell surface. This overview emphasizes the phosphoinositidase C mediated dual messenger pathway in brain and in brain-derived cells, with special reference to possible significance for research on the dementias. PMID- 8239266 TI - Production of amyloid beta protein from normal amyloid beta-protein precursor (beta APP) and the mutated beta APPS linked to familial Alzheimer's disease. AB - The approximately 4 kD (39-43 amino acid) polypeptide (amyloid beta protein, A beta) deposited as amyloid in Alzheimer's disease (AD) is derived from a set of 695-770 residue precursor proteins collectively referred to as the amyloid beta protein precursor (beta APP). Using immunoblotting techniques, metabolic labeling, and sequencing we have analyzed beta APP derivatives in medium conditioned by: (1) human mononuclear leukemic (K562) cells expressing a model beta AP-bearing carboxyl-terminal beta APP derivative (2) human neuroblastoma (M17) cells transfected with constructs expressing full length beta APP and (3) M17 cells expressing only endogenous beta APP. In each case, we observed the release of a approximately 4 kD beta APP derivative essentially identical to the A beta found in AD amyloid. A similar, if not identical, beta APP fragment was readily detected in CSF from both Alzheimer's disease patients and controls. These observations indicate that the A beta is produced and released by normal processing of the beta APP. To determine if the production of A beta or A beta tearing COOH-terminal beta APP derivatives is altered in cells expressing the mutant beta APPs linked to familial AD, we have compared M17 cells expressing wild type beta APP with those expressing mutant beta APPs (beta APP delta I or beta APP delta NL). After continuous metabolic labeling for 8 hours, cells expressing the beta APP delta NL mutant showed a 5-fold increase in the relative amount of an approximately 11.4 kD A beta-bearing carboxyl-terminal beta APP derivative, and they released 6-fold more 4 kD A beta into the medium. These observations provide strong evidence that: (1) the pathway producing A beta in cultured cells is highly relevant to AD and (2) the beta APP delta NL mutant causes AD because its processing is altered in a way that releases increased amounts of A beta. PMID- 8239267 TI - Normal cellular processing of the beta-amyloid precursor protein results in the secretion of the amyloid beta peptide and related molecules. AB - Alzheimer's disease is characterized by the extracellular deposition in the brain and its blood vessels of insoluble aggregates of the amyloid beta peptide (A beta). This peptide is derived from a large integral membrane protein, the beta amyloid precursor protein (beta APP), by proteolytic processing. The A beta has previously been found only in the brains of patients with Alzheimer's disease or advanced aging. We describe here the finding that A beta is produced continuously by normal processing in tissue culture cells. A beta and closely related peptides were identified in the media of cells transfected with cDNAs coding for beta APP in a variety of cell lines and primary tissue cultured cells. The identity of these peptides was confirmed by epitope mapping and radiosequencing. Peptides of a molecular weight of approximately 3 and approximately 4 kDa are described. The 4 kDa range contains mostly the A beta and two related peptides starting N terminal to the beginning of A beta. In the 3 kDa range, the majority of peptides start at the secretase site; in addition, two longer peptides were found starting at amino acid F(4) and E(11) of the A beta sequence. To identify the processing pathways which lead to the secretion of these peptides, we used a variety of drugs known to interfere with certain cell biological pathways. We conclude that lysosomes may not play a predominant role in the formation of 3 and 4 kDa peptides. We show that an acidic environment is necessary to create the N terminus of the A beta and postulate that alternative secretory cleavage might result in the formation of the N-terminus of A beta and related peptides. This cleavage takes place either in the late Golgi, at the cell-surface or in early endosomes, but not in lysosomes. The N-terminus of most of the 3 kDa peptides is created by secretory cleavage on the cell surface or within late Golgi. PMID- 8239268 TI - Protein phosphorylation regulates relative utilization of processing pathways for Alzheimer beta/A4 amyloid precursor protein. AB - The Alzheimer amyloid precursor protein (APP) is a phosphoprotein, and the phosphorylation state of APP at Ser655 can be regulated by protein kinase C, calcium/calmodulin-dependent protein kinase II, and okadaic acid-sensitive protein phosphatases. Other enzymes may also play a role at Ser655 of APP and, perhaps, at other residues. Signal transduction via protein phosphorylation regulates APP metabolism. In particular, APP processing via the nonamyloidogenic secretory cleavage pathway is increased following the activation of protein kinase C or the inactivation of okadaic acid-sensitive protein phosphatases. The mechanism(s) by which protein phosphorylation regulates APP secretory cleavage include (among others): substrate activation, substrate redistribution, protease activation and/or protease redistribution. Current experimental evidence will be discussed, addressing the relative importance of each of these possibilities and the implications for these events in the modulation of beta/A4-amyloidogenesis. PMID- 8239269 TI - Receptor-coupled amyloid precursor protein processing. AB - The family of beta-amyloid protein precursors (APP) can be processed via several alternative proteolytic pathways. Some generate potentially amyloidogenic APP derivatives, whereas others preclude the formation of such fragments. The cellular mechanisms regulating the relative activities of these pathways are thus important in determining the factors contributing to the formation of amyloidogenic APP derivatives. In order to investigate whether cell-surface receptor activity can regulate APP processing, HEK 293 cell lines stably expressing human muscarinic acetylcholine receptors (mAChR; subtypes m1, m2, m3, m4) were stimulated with the muscarinic agonist carbachol, and the release of APP derivatives was measured. Carbachol increased the release of large amino-terminal APP-fragments 4- to 6-fold in cell lines expressing the m1 or m3 receptors but not in those expressing m2 or m4 subtypes. This increase was blocked by various protein kinase inhibitors and mimicked by phorbol esters, indicating that it is mediated by protein kinase activation, presumably by protein kinase C (PKC). To determine whether additional cell-surface receptor types linked to this signal transduction pathway could also regulate APP processing, we stimulated differentiated PC-12 cells with bradykinin and found that this neuropeptide also increased the secretion of amino-terminal APP derivatives. We next investigated the possibility that neuronal depolarization might affect APP processing in mammalian brain. Electrically stimulated rat hippocampal slices released two times more amino-terminal APP derivatives than unstimulated control slices. This release increased with increasing stimulation frequencies in the physiological firing range of hippocampal pyramidal cells, and was blocked by tetrodotoxin. These results suggest that, in brain, APP processing is regulated by neuronal activity. PMID- 8239270 TI - Regulation of amyloid precursor protein release by protein kinase C in Swiss 3T3 fibroblasts. AB - Release of the amyloid precursor protein (APP) of Alzheimer's disease from Swiss 3T3 fibroblasts was stimulated in a concentration-dependent manner by phorbol 12 myristate 13-acetate. In fibroblasts overexpressing protein kinase C alpha (PKC alpha), the EC50 for this response was 7 nM, while in control cells the EC50 was 63 nM. The effect of PMA was inhibited by the PKC antagonist H-7 in control cells, but not in cells that overexpressed PKC alpha. Basal release of APP was higher in cells that overexpressed PKC alpha, and was not affected by the phosphatase inhibitor okadaic acid, although this compound doubled APP release from control cells. The results suggest that PKC alpha regulates APP processing in mammalian cells. Alterations in the activity of PKC have been reported to occur in Alzheimer's disease and might potentially contribute to abnormalities of APP metabolism characteristic of this disorder. PMID- 8239271 TI - Cellular processing and proteoglycan nature of amyloid precursor proteins. AB - Amyloid beta protein (beta/A4 or A beta), the main proteinaceous component of the amyloid depositions of the Alzheimer's brain, derives from the proteolytic processing of the amyloid precursor protein (APP). Cleavage of the amyloid precursor by at least two distinct secretase activities produces soluble secreted APP. The major secretase cleavage (site I) takes place between A beta 16 and 17, while the minor cleavage (site II) takes place after A beta Lys 28 and may produce potentially amyloidogenic secreted APP. Full-length cellular APP is cleaved by secretase intracellularly in the Trans-Golgi Network (TGN) or in post Golgi vesicles. The resultant soluble APP is transported to the plasma membrane and exocytosed. The biological activity of the APP is still not completely understood, although it seems to act as a cell adhesion molecule. Recent studies have shown that in glioma cells, most of the soluble secreted APP occurs as a chondroitin sulfate proteoglycan (CSPG). In addition, full length APP CSPG has been detected in neuroblastoma and fibroblast cells as well as on the surface of glioma cells, and in human brain. These results suggest that the proteoglycan nature of the APP proteins may be important for their biological function. PMID- 8239272 TI - Sequestration of amyloid beta-peptide. AB - Amyloid beta-protein, or beta/A4, is a 4-kilodalton peptide that forms poorly soluble extracellular depositions of amyloid in brains and leptomeninges of patients with Alzheimer's disease (AD), Down's syndrome (DS), and hereditary cerebral hemorrhage with amyloidosis-Dutch type (HCHWA-D). beta/A4 peptide is a derivative of a large transmembrane glycoprotein (APP) and is found in the extracellular space, i.e., in the cerebrospinal fluid and serum of individuals with and without AD and in the conditioned media of many different cells grown in culture. The mechanism by which normally produced amyloid beta peptide forms extracellular aggregates in patients is unknown. One possible explanation is a failure of a mechanism for removal of the beta/A4 peptide that prevents this highly aggregating peptide from forming extracellular amyloid depositions. PMID- 8239273 TI - The C-terminus of the beta protein is critical in amyloidogenesis. AB - The beta amyloid protein found in extracellular deposits in Alzheimer's disease (AD) is heterogeneous at its C-terminus; proteins ending at residues 40, 42, and 43 have been identified in neuritic deposits, while protein in vascular amyloid appears to end at residue 39 or 40. Studies of synthetic beta proteins (beta 1 39, beta 1-40, beta 1-42), and model peptides (beta 26-39, beta 26-40, beta 26 42, beta 26-43) demonstrate that amyloid formation is a nucleation-dependent phenomenon. Peptides ending at residues 39 or 40 were kinetically soluble for hours to days, while peptides ending at residues 42 or 43 aggregated immediately; all eventually reached similar thermodynamic solubility. The kinetically soluble variants could be seeded with the kinetically insoluble variants. The secondary structure of beta 26-39 fibrils was different from that of beta 26-42 fibrils, however, seeding beta 26-39 with beta 26-42 produces mixed fibrils with structure similar to beta 26-42. These results suggest that neuritic plaques may be seeded by their minor component; this may determine the structure and properties of amyloid in AD. PMID- 8239274 TI - Biologically active domain of the secreted form of the amyloid beta/A4 protein precursor. AB - The amyloid beta/A4 protein precursor (APP), a large transmembrane protein, is expressed ubiquitously in many organisms, as well as in a variety of cultured cells. Studies of the synthesis and processing of APP have revealed several intricate metabolic pathways for this protein. One of these pathways involves the cleavage of APP in the middle of the beta/A4 domain and results in the secretion of the large amino-terminal portion of the protein. The biological function of this secreted form of APP has been the subject of intense investigation by several groups and various activities have been described for the different domains of APP studied. Our initial approach was to create a fibroblast cell line in which APP expression is dramatically reduced. These fibroblasts, called A-1, have a very slow growth rate. Addition of exogenous APP in the medium of A-1 cells restores their growth to the level of normal parent fibroblasts, demonstrating a growth factor-like activity for the secreted form of APP. Using APP fragments made in bacteria as well as synthetic peptides, we have been able to locate the active site of APP within a domain of 17 amino-acids (Ala319 Met335). This domain of APP can stimulate neurite extension of cultured neuroblastoma cells and it is proposed that APP mediates this effect through binding to a cell surface receptor, triggering intracellular transduction mechanisms. Thus, the secreted form of APP can function as a growth and/or differentiation factor and the site involved in these activities is within a 17 mer domain in the middle of the molecule. Our current lines of research seek to further characterize the mechanisms of APP function as well as its activity in vivo. PMID- 8239275 TI - Muscarinic acetylcholine receptor subtypes associated with release of Alzheimer amyloid precursor derivatives activate multiple signal transduction pathways. AB - Five subtypes of muscarinic acetylcholine receptors have been identified and designated m1-m5. The m1 and m3 receptors have recently been shown to stimulate APP processing. The m1 and m3 receptors couple to a variety of signal transduction pathways in both tissue slices and a variety of cell lines endogenously expressing either or both subtypes. In contrast, the m2 and m4 receptors have been primarily associated with inhibition of adenylate cyclase. We have transfected all five subtypes of muscarinic receptors into a variety of mammalian cell lines in order to investigate the signaling associated with single receptor subtypes. The m1, m3, or m5 receptors stimulate phospholipase A2, C, and D, adenylate cyclase, receptor-operated calcium channels, and tyrosine kinase activity simultaneously. The m2 or m4 receptor inhibits cAMP accumulation and augments a previously stimulated release of arachidonic acid and calcium influx. PMID- 8239276 TI - beta-Amyloid precursor protein mismetabolism and loss of calcium homeostasis in Alzheimer's disease. AB - The suspected involvement of the beta-amyloid precursor protein (beta APP) in the etiology of Alzheimer's disease (AD) has been strengthened by recent genetic evidence, but pursuit of the mechanisms involved will initially require basic cell biology approaches. Several studies have concentrated on toxic activities of beta-amyloid peptide (beta AP) itself, illuminating its contributions to excitotoxicity and calcium-mediated degeneration in general. We now know that generation of beta AP from beta APP also compromises the production of an important set of trophic factors: the secreted forms of beta APP (APPS), which may act--ironically--by conferring protection from calcium-mediated insults. Therefore, conditions which contribute to the formation of beta AP (possibly including ischemia) not only produce an agent which exacerbates calcium-mediated cell death, but also reduce the levels of one of the few factors able to rescue calcium homeostasis. The implications of these postulates and their relationship to the process of aging are discussed. PMID- 8239277 TI - A new hypothesis for the mechanism of amyloid toxicity, based on the calcium channel activity of amyloid beta protein (A beta P) in phospholipid bilayer membranes. AB - Amyloid beta protein (A beta P) is the 40-42 residue polypeptide implicated in the pathogenesis of Alzheimer's disease (AD). We have reconstituted this peptide into phosphatidylserine liposomes and then fused the liposomes with a planar lipid bilayer. When incorporated into this bilayer, the A beta P forms cation selective channels capable of transporting calcium and some monovalent cations including cesium, lithium, potassium, and sodium. The channels behave in an ohmic fashion and single channels can be shown to exhibit multiple subconductance states. Hitherto, A beta P has been presumed to be neurotoxic, although direct demonstration of toxicity has proved elusive. On the basis of the present data we suggest that the ion channel activity of the polypeptide may be the basis of its neurotoxic effects. PMID- 8239278 TI - The role of extracellular matrix in the processing of the amyloid protein precursor of Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by the presence of extracellular amyloid plaques, which contain a protein referred to as the amyloid or beta A4 protein. The beta A4 protein is derived from a larger precursor protein (APP). Studies of autosomal-dominant forms of AD have established the central role of APP in the pathogenesis of the disease. Despite considerable research, the function of APP is unknown. APP can be processed by at least two separate routes. The first route involves a protease known as "APP secretase," which cleaves within the amyloid sequence, thereby mitigating amyloid formation. The second route may result in the production of potentially amyloidogenic fragments. Our studies suggest that following release from the cell membrane, APP interacts with components of the extracellular matrix (ECM) such as the heparan sulfate proteoglycans (HSPG's). The interaction of APP with HSPG's may be important for the function of APP. Substratum-bound APP was found to dramatically increase neurite outgrowth and survival of chick sympathetic neurons in vitro. This effect was dependent upon the presence of substratum-bound HSPG. The results suggest that normally, when bound to the ECM, APP functions to promote neurite outgrowth and/or cell survival. Loss of this normal trophic function might occur in AD, when APP is proteolytically processed via the amyloidogenic pathway. PMID- 8239279 TI - The beta A4 amyloid protein precursor in human circulation. AB - beta A4, the principal constituent of the brain amyloid collections in Alzheimer's disease, is derived from a much larger precursor, the amyloid protein precursor (APP). APP exists in the blood as full-length, potentially amyloidogenic forms in platelets, and as an attenuated species in plasma and T lymphocytes. Studies of circulating APP facilitate the elaboration of the function of this protein, as well as the elucidation of its processing in health and disease. PMID- 8239280 TI - Expression of L-APP mRNA in brain cells. AB - Several reports addressed the issue of how the alternative splicing of exon 7 and 8 in the APP pre-mRNA is regulated in different tissues. Of special interest here was the potential involvement of exon 7 containing APP splice isoforms, since this exon codes for a serine protease inhibitor and is therefore of putative relevance for amyloidogenic catabolism of the precursor protein. The recent identification of a third alternative splice site in close proximity to the beta A4-amyloid portion in the APP gene which may also increase APP amyloidogenicity, allowed us to investigate its regulation in cells of the central nervous system. With our assay, we were able to resolve six different APP isoforms of the eight potential isoforms which can be generated from the three alternatively spliced exons 7, 8, and 15. We demonstrate here that, in addition to rat brain microglia cells, astrocyte-enriched cultures also skip the novel alternative 3'-splice site in front of exon 15, generating L-APP mRNA. Neurons are the only cells in the central nervous system which seem to use the 3'-splice site of intron 14 nearly 100%. Interestingly, this very 3'-splice site is the only one present in the APP gene that completely matches the consensus sequence for the branchpoint sequence proposed for introns. We would therefore suggest that neurons lack a specific splicing factor which inhibits the use of the rather strong 3'-splice site in front of exon 15. It remains to be shown whether this is also the case for neurons in Alzheimer's disease. PMID- 8239281 TI - The amyloid precursor protein in ischemic brain injury and chronic hypoperfusion. AB - We studied changes in the spatial and temporal distribution of the beta amyloid precursor protein (APP) of Alzheimer's disease (AD) in experimental ischemic brain injury. Rats with repeated reversible occlusions of one middle cerebral artery showed striking APP reactivity in astrocytic processes in perifocal regions and adjacent white matter. APP reactive dystrophic axons and neurons were also evident in the cortex and hippocampus ipsilateral to the MCA occlusion. Such changes were similarly apparent in animals subjected to partial forebrain ischemia induced by bilateral occlusion of the carotid arteries. Our studies suggest that focal ischemic insults or chronic hypoperfusion leads to increased accumulation or induction of APP in surviving cellular elements that may relate to the processes involved in beta amyloid deposition in AD. PMID- 8239282 TI - Heat-shocked neuronal PC12 cells reveal Alzheimer's disease--associated alterations in amyloid precursor protein and tau. AB - The Alzheimer's disease (AD) brain contains many abnormal protein modifications. These include the abnormal processing of amyloid precursor protein (APP) to form the amyloidogenic beta/A4 peptide and the abnormal phosphorylation of tau to form A68, the major constituent of the neurofibrillary tangle. In addition, many of the biochemical alterations found in the AD brain are also found in heat-shocked or stressed cells. We used heat-shocked neuronal PC12 cells to investigate the effects of stress on APP and tau. We found that by simply exposing neuronal PC12 cells to an elevated temperature (45 degrees C) for 30 minutes, they exhibited several features characteristic of the heat shock response. These included a 45% reduction in total protein synthesis, the induction of heat shock protein (hsp) 72, and increased phosphorylation of the protein synthesis initiation factor eIF 2 alpha. The heat-shocked cells also exhibited alterations in the metabolism and phosphorylation of APP. Under heat shock conditions, we found two additional APP like polypeptides not present in controls and a significant decrease in the phosphorylation state of APP. We also found that an A68-like protein is formed in neuronal PC12 cells when subjected to elevated temperature. This A68-like protein was formed with heat shock even in the absence of protein synthesis, suggesting that its production occurred post-translationally. The tau/A68 polypeptides were identified as phosphoproteins, and the phosphorylation of tau to form A68 was reversed with recovery of the cells from heat shock. Immunoprecipitation of lysates from heat shocked cells with antibodies to hsp72/73 resulted in co precipitation of tau, but not A68 with hsp72 indicating a stable complex formation between these two proteins. These results suggest that heat shock proteins may play either a protective or promoting role in the formation of A68 and/or the amyloidogenic C-terminal fragment of APP. PMID- 8239283 TI - Alzheimer's disease families with amyloid precursor protein mutations. AB - Early onset Familial Alzheimer's Disease (FAD) is an autosomal dominant disease with apparent complete penetrance. It is genetically heterogeneous with some families carrying mutations in the amyloid precursor protein (APP) gene which segregate with the disease. In addition, there is allelic heterogeneity with four mutations associated with FAD. Three mutations have been reported at APP 717, just distal to the C-terminus of the beta-amyloid domain, APP 717 val-ile, APP 717 val-phe, and APP 717 val-gly, which are associated with autopsy-proven Alzheimer's disease (AD). APP 670/671 lies at the N terminus of the beta-amyloid domain and is associated with clinically diagnosed FAD in two Swedish families. FAD tends to have prominent myoclonus and this is shared by the cases with APP mutations. In two unrelated UK families with APP 717 val-ile mutations there was early prominent memory impairment with dyscalculia proceeding to generalized cognitive impairment with a lack of insight. There was a late development of a gait disturbance with extrapyramidal features in some members. Positron emission tomography (PET) with fluorodeoxyglucose demonstrated posterior bitemporal biparietal hypometabolism in one case. Magnetic resonance imaging (MRI) showed generalized cerebral atrophy particularly affecting the temporal lobes and hippocampus. At autopsy, a single case showed extensive beta-amyloid deposition with congophilic angiopathy and widespread senile plaques and neurofibrillary tangles. The cytoskeletal pathology associated with abnormally phosphorylated tau was similar to cases of sporadic AD. In addition, there were widespread cortical and subcortical Lewy bodies. A single family with the APP 717 val-gly mutation also showed prominent myoclonus, lack of insight, and seizures, PET, in a single case, showed classical biparietal bitemporal hypometabolism. Autopsy, in a single case, showed diffuse deposits of beta-amyloid throughout the cortex with frequent neuritic plaques and neurofibrillary tangles. No other inclusion bodies were seen. There was severe congophilic angiopathy. The age at onset of APP mutations is around 50 years of age by contrast to other early onset FAD pedigrees. PMID- 8239284 TI - Search for the genes responsible for familial Alzheimer's disease. AB - Inherited or Familial Alzheimer's Disease (FAD) has clearly been shown to be a genetically heterogeneous disorder. Mutations in the gene on chromosome 21 encoding the beta-amyloid protein precursor (APP) have been shown to be linked to 2-3% of FAD kindreds examined around the world. A late onset FAD locus has been mapped to a region of chromosome 19 in which a recently isolated APP-like gene, APLP1 has also been localized, making this gene a strong candidate to harbor a late-onset FAD defect. More recently, a major FAD locus has been mapped to the long arm of chromosome 14. The chromosome 14 locus appears to be mainly linked to the gene defect in early onset FAD pedigrees. Besides the FAD loci on chromosome 21, 19, and 14, at least two other loci must exist since the gene defect in some early- and late-onset FAD pedigrees do not appear to segregate with markers from any of these autosomes. As different gene defects responsible for various forms of FAD are discovered, perhaps, a common basis for the etiology of this devastating disorder can be discerned. PMID- 8239285 TI - Transgenic animal models for Alzheimer's disease. AB - The neuropathology of Alzheimer's disease is characterized by the deposition of abnormal protein aggregates. The main constituent of the deposition is beta amyloid protein. A seminal role of this protein is supported by the discovery of point mutations in the gene of its precursor protein in certain forms of familial Alzheimer's disease. In vitro (cultured neuronal cells), overexpression of the precursor protein or a part of the precursor leads to degeneration of neurons, suggesting neurotoxicity of its derivatives. At this time, all of the reported transgenic mice bearing DNA construct for the precursor or a part of the precursor, however, have not developed convincing pathological changes similar to what is observed in patients with Alzheimer's disease. This interesting discrepancy between in vitro and in vivo suggests suppressors in vivo which ameliorate beta-amyloid precursor protein derivative-mediated neurotoxicity. PMID- 8239286 TI - Transgenic mice expressing human beta-APP751, but not mice expressing beta APP695, display early Alzheimer's disease-like histopathology. AB - Mice transgenic for the 751 amino acid isoform of the human beta-amyloid precursor protein (beta-APP) driven by the rat neuron specific enolase (NSE) promoter (NSE:beta-APP751) show features of early Alzheimer's disease (AD) pathology. These features, which were evident in multiple pedigrees, include: 1) preamyloid deposits which stain with antibodies that are specific for the beta amyloid peptide and stain AD amyloid deposits and plaques, and 2) neuronal soma and processes which stain with an antibody (Alz50) that detects abnormal isoforms of tau which are characteristic of AD. The quality and distribution of both types of immunoreactivity revealed in the NSE:beta-APP751 mouse brains most closely resemble those seen in brains of young adults with Down's syndrome. Both structures are rarely, if ever, observed in brains from mice transgenic for the 695 amino acid isoform of beta-APP (NSE:beta-APP695) or in wild type mice. PMID- 8239287 TI - Experimental induction of beta-amyloid plaques and cerebral angiopathy in primates. AB - Moderate numbers of amyloid plaques with associated argyrophilic dystrophic neurites and cerebral amyloid angiopathy (CAA) but no neurofibrillary tangles (NFTs) were found in the brains of 3 middle-aged common marmosets (Callithrix jacchus) inoculated intracerebrally (i.c.) 6-7 years earlier with brain tissue from a patient with early onset Alzheimer's disease. The plaques and vascular amyloid stained positively with antibodies to beta (A4)-protein. The brains of 3 age-matched control marmosets from the same colony did not show these neuropathological features. beta-amyloid plaques and CAA (but no spongiform encephalopathy) were also found in the brain of a marmoset inoculated with brain tissue from a patient with prion disease with concomitant beta-amyloid plaques and CAA. An occasional beta-amyloid plaque was found in the brains of two marmosets inoculated with brain tissue from elderly patients. No beta-amyloid plaques nor CAA were found in 6 other marmosets who were older than the inoculated marmosets, 10 further marmosets who were slightly younger but who had been inoculated several years previously with brain tissue which did not contain beta-amyloid, and 10 younger marmosets who had been subjected to various neurosurgical procedures. These results suggest that beta-amyloidosis is a transmissible process. PMID- 8239288 TI - The age of biosenescence and the incidence of cerebral beta-amyloidosis in aged captive rhesus monkeys. AB - Autopsy surveillance of 186 rhesus monkeys aged 20 to 36 years revealed that development of major geriatric diseases such as emphysema, coronary sclerosis, and cancer increased rapidly after the age of 25 years, and nearly 70% of the monkeys in each cohort group died by 30 years. According to our 12-year longitudinal survey, the age of biosenescence in captive rhesus monkeys begins around 25 years and the maximum longevity is 36 years. The incidence of cerebral beta-amyloidosis associated with plaque formation and cerebral angiopathy was observed in 51 brains of rhesus monkeys aged 25 to 36 years. Lesions were found in 31 of 51 aged brains (60%) and 6 monkeys over 34 years of age were all severely affected. Despite the size of the plaque, nearly all of them showed immunopositive beta-amyloid. Cerebral angiopathy coexisted in 10 of 31 plaque positive brains. The basal prefrontal gyrus was the most common site and contained the highest density of plaques, followed by the amygdala region. The amyloid in the liver, spleen, adrenal and pancreatic islets in visceral amyloidosis showed no positivity to the beta-amyloid demonstrated in the brain. As in aged human brains, the incidence of age-dependent cerebral beta-amyloidosis in captive rhesus monkeys showed great individual variation. PMID- 8239289 TI - Cholinergic and non-cholinergic synaptic mechanisms in the aged rat hippocampus. AB - We compared age-related alterations in the electrophysiological and pharmacological properties of CA1 hippocampal pyramidal neurons in three strains of rats (Sprague-Dawley, Fisher 344 and Wistar) at 3-4 and 25-32 months of age, using the in vitro slice preparation. The most consistent age-related alterations were: a decrease in membrane excitability, a decrease in the amplitude and duration of inhibitory postsynaptic potentials and a decreased sensitivity to the effect of the cholinergic agonist carbachol. In contrast, no consistent alterations in calcium-dependent events were observed in these strains of rats. The age-related changes in the duration of the after-hyperpolarization (AHP) were different (and even opposite) depending on the strain studied. Our results show that age-related changes observed in a given strain are not necessarily present in all strains of the same species. PMID- 8239290 TI - Phospholipid and phospholipid metabolites in rat frontal cortex are decreased following nucleus basalis lesions. AB - Membrane phospholipid metabolism is abnormal in Alzheimer's disease (AD) brain. Phosphatidylcholine and phosphatidylethanolamine levels are decreased as are choline and ethanolamine, while glycerophosphocholine (GPC) and glycerophosphoethanolamine are increased. To develop a rat model for these changes, we examined the effects of unilateral lesion of the cholinergic nucleus basalis (nBM) with ibotenic acid (10 mg/ml in PBS, 0.5 microliter) and sham lesion on frontocortical phospholipid, choline and GPC. After one week, choline acetyltransferase activity in frontal cortex was decreased (26%, p < 0.005, n = 14) on the nBM ibotenate-lesion side relative to the contralateral side, while there were no differences following the nBM sham-lesion. Levels of membrane phospholipids (nmol/mg protein) in adjacent frontal cortex sections exhibited concomitant decreases (13%, p < 0.05, n = 14) on the nBM ibotenate-lesion side, while there were no differences following the nBM sham-lesion. Tissue nBM ibotenate-lesion frontocortical choline and GPC levels were also decreased relative to those in control tissue (choline: 21%, p < 0.05, n = 14; GPC: 10%, p < 0.05, n = 14), while nBM sham-lesion showed no effect. Muscarinic receptor sensitivity in frontal cortex following nBM ibotenate-lesion was increased, as measured by carbachol-stimulated inositol phosphate production (p < 0.001, n = 12), indicating that increased receptor mediated phospholipid hydrolysis in cortex may occur following nBM ibotenate-lesion. These data suggest that impaired cholinergic transmission alters phospholipid metabolism in cholinergic target regions. PMID- 8239291 TI - Decreased density of forebrain cholinergic neurons and disintegration of the spatial organization of behavior in experimental autoimmune dementia (EAD). AB - Experimental autoimmune dementia (EAD) is a rat model designed to examine the potential role of anti-cholinergic neurons antibodies (Abs) in the neuropathology of Alzheimer's disease (AD) and dementia. We have previously shown that sera of AD and Down's syndrome patients contain Abs which bind specifically to the high molecular weight neurofilament protein (NF-H) of the purely cholinergic electromotor neurons of Torpedo. Production of such Abs in EAD rats by prolonged immunization with Torpedo cholinergic NF-H results in the accumulation of IgG in the septum and hippocampus of the immunized rats and in memory deficits. In the present study, we examined immunohistochemically whether the anti-cholinergic NF H immune response of the EAD rats affects their brain cholinergic neurons. In addition, since dementia is associated with severe deterioration in the spatio temporal organization of behavior, we examined whether EAD rats also mimic this important feature of dementia. The results obtained show that production in EAD rats of anti-cholinergic NF-H Abs similar to those found in AD patients results in a marked decrease in the density of forebrain cholinergic neurons and in derangements in the spatio-temporal organization of their behavior. These findings may replicate pathogenic processes in AD and support a role for anti cholinergic NF-H Abs in the degeneration of cholinergic neurons in the disease. PMID- 8239292 TI - Age-related cognitive deficits in rats are associated with a combined loss of cholinergic and serotonergic functions. AB - The cholinergic hypothesis of senile dementia proposes that an age-dependent reduction of central cholinergic functions accounts for the severe cognitive deficits seen in aged rats. A careful examination of the experimental evidence cited in support of this hypothesis reveals that it cannot account for some behavioral observations. We have modified this hypothesis and wish to propose that serotonin and acetylcholine interact to allow normal cognitive functions in the brain. Thus, a partial reduction in both cholinergic and serotonergic functions will cause severe memory impairment in young as well as in aged rats. We found that restoration of the serotonergic innervation in the hippocampus of serotonin depleted rats, using tissue transplants, can restore impaired behavior. We have localized a memory-related interaction between serotonin and acetylcholine in the hippocampus and are in the process of identifying a physiological function which may underly this interaction. PMID- 8239293 TI - Neural transplantation for neurodegenerative diseases: past, present, and future. AB - After almost 100 years of sporadic, and marginally successful, studies of neural transplantation in animals, we are now on the threshold of a clinical treatment of the damaged brain. The initial studies of neural transplantation have focused on Parkinson's disease, primarily as a model for a more general strategy of "repair by cellular replacement." Parkinson's is known to result from the loss of a small population of cells that produce the essential neuromodulator, dopamine, for much of the brain. Further, the disease is improved significantly, during the early part of its course, by chemical augmentation of dopamine activity through drug therapies, such as L-dopa. Finally, the disease is often fatal in spite of the best medical treatments, therefore justifying more radical therapeutic experiments. If transplantation of brain cells can be accomplished successfully in humans, as it has been in animals, then replacement of a small population of dopamine-producing cells in Parkinson's disease should have important functional effects and possibly reverse the course and symptoms of the disease. Other useful applications will surely follow for conditions affecting millions of people for whom medicine now has only palliative and ineffective treatments. Just as Parkinson's disease is a model clinical condition for testing cellular replacements, fetal neural tissue transplants are also a first step for a broader strategy of molecular and cellular therapies. Fetal cells are, in many respects, the best replacements one could imagine, since precursor cells have the capacity to develop into every cell found in the adult. So, the best replacement for a dopamine neuron would likely be a precursor dopamine neuron or "neuroblast." Animal research through 1985 had demonstrated the unique properties of such fetal cells, but survivability after transplantation had not been attained with primate or human neural tissue. Our programs developed techniques to transplant monkey fetal neural tissue, to cryopreserve it, and to reverse functional effects of the neurotoxin, MPTP, in monkeys. This technique was applied to the collection and preservation of human tissue, and preliminary successful results have been obtained in patients with idiopathic Parkinson's disease. Others have reported success with different techniques in two MPTP-Parkinsonian patients and a small number of patients with idiopathic disease. If the most dramatic improvements can be replicated consistently and the benefits last for a reasonable period without complications, a clinical treatment might develop using "random-source" fetal cadaver cells. PMID- 8239294 TI - Basal forebrain grafts in the hippocampus and neocortex: regulation of acetylcholine release. AB - The regulation of acetylcholine (ACh) release from cholinergic neurons transplanted to the hippocampus or neocortex was studied by microdialysis in awake rats. Fetal basal forebrain tissue was implanted as a cell suspension or solid graft into the fimbria-fornix-lesioned hippocampus, or as a cell suspension into the frontal cortex after excitotoxic lesion of the nucleus basalis. Several months after transplantation, microdialysis probes were implanted in areas of the hippocampus or frontal cortex reinnervated by the grafts. The grafts restored lesion-induced deficits in steady-state ACh release up to normal or above normal levels in both hippocampus and frontal cortex. The responses to KCl and tetrodotoxin suggested that the ACh release exhibited normal firing-dependent properties. By applying various behaviorally arousing stimuli that normally activate the basal forebrain projection systems, we wished to investigate the functional integration of the grafts in the host brain. In the hippocampus, sensory stimulation, immobilization stress and motor activity all resulted in increased release of graft-derived ACh amounting to 25-65% of the normal response. Variations in ACh levels during the day-night cycle was, however, not observed in the grafted rats. In the frontal cortex, immobilization enhanced the graft-derived ACh release (60% of normal response), whereas the response to sensory stimulation did not reach significance. Since the activity of the normal basal forebrain projection systems is under influence of monoaminergic brainstem afferents, we investigated the effects of systemic administration of amphetamine or apomorphine on ACh release in the hippocampus. Both drugs produced increases in graft-derived ACh release although the response was variable and less pronounced than normal. In conclusion, the graft-derived ACh release was affected by behavioral manipulations and catecholaminergic drugs that normally modify cholinergic septo-hippocampal and basalo-cortical activity. This strongly suggests a high degree of functional integration of the graft in the host brain allowing for a regulated release of transmitter that can be adjusted during ongoing behavior. PMID- 8239295 TI - Behavioral effects of cholinergic grafts. AB - Experimental work in animals and, to a more limited extent, in humans, has demonstrated that the cholinergic system is involved in mechanisms which control learning and memory. Since there is cholinergic loss in a variety of dementing illnesses, any treatment designed to alleviate the mental symptoms of these diseases must address the issue of cholinergic dysfunction even if other treatments are also required to overcome other neurotransmitter imbalances. Work in rodents has demonstrated that cholinergic-rich fetal neural tissue transplants can, under certain circumstances, alleviate the behavioral effects of cholinergic lesions or of cholinergic decline associated with aging. More complex cognitive testing can be achieved using primates and, in this case, the common marmoset is a suitable species to use because its rapid and reliable reproductive rate aids the provision of appropriate transplant tissue. Marmosets with transection of the fornix are deprived of a cholinergic input into the dentate gyrus, posterior hippocampus and entorhinal cortex and are specifically impaired on learning tasks which require remembering a rule of responding (non-evaluative memory). Transplantation of cholinergic-rich fetal septal tissue into the hippocampus of such animals completely restores their ability to learn this type of task, whereas transplantation of cholinergic-poor fetal hippocampal tissue into the same area produces no such improvements. These results demonstrate that where a learning impairment is produced by a relatively simple procedure which has a major effect on one neurotransmitter, that function can be restored by transplantation of tissue containing that neurotransmitter even where the impairment consists of a very "high level" cognitive dysfunction. PMID- 8239296 TI - Cells engineered to produce acetylcholine: therapeutic potential for Alzheimer's disease. AB - Alzheimer's disease (AD) is a debilitating disorder of the central nervous system which may affect up to 50% of the population over the age of 85 years. The etiology of AD is unknown and there is currently no cure for the disease. Well documented losses in cholinergic and other neurotransmitter systems have provided a focal point for attempting pharmacological interventions in AD to ameliorate some of the cognitive deficits that occur. However, current systemic strategies have met with limited success. An alternative strategy, that has been pursued in animal models of neurodegenerative disease, is to augment neurotransmitter function within the brain through tissue transplantation. Such implants have an advantage over conventional drug therapies in that the cells can be precisely placed within compromised areas of the brain. We have pursued a strategy of designing cells, through the use of molecular biology techniques, to produce neurotrophic factors and neurotransmitters. Recently, we developed a primary fibroblast cell line that was genetically modified to express choline acetyltransferase (ChAT). In vitro, these cells produced and released acetylcholine at levels that varied with the amount of choline in the culture media. When implanted into the hippocampus of rats, the in vivo microdialysis technique revealed that the ChAT-expressing fibroblasts continued to produce and release acetylcholine after grafting. Most importantly, the levels of acetylcholine synthesized by the cells could be regulated by the localized infusion of choline in the vicinity of the grafts. These results confirmed previous work which indicated that engineered fibroblasts provide an effective delivery vehicle of different substances to the brain. While the intracerebral implantation of genetically modified cells will not cure AD, the continuing development of this strategy may ultimately provide a powerful approach for ameliorating the devastating cognitive impairments which are a hallmark of this disease. PMID- 8239297 TI - Neural stem cells for CNS transplantation. AB - Neurodegenerative disorders such as Parkinson's, Alzheimer's, and Huntington's disease are becoming ever more prominent in our society. A direct approach towards therapeutic treatment of these diseases is through replacement therapy where normal tissue is transplanted back to the nervous system. Recently, significant progress has been achieved with transplants in Parkinson's disease, but the process is heavily dependent on an unstable and problematic source of fetal tissue. Neural stem cells may become the tissue/cell source necessary for developing the therapeutic potential of neural transplantation. Stem cells are self-renewing, multipotent and could provide a well-characterized and clean source of transplantable material. A number of new in vitro approaches have led to the development of continuously propagated stem cells that are potential candidates for nervous system transplantation. These include oncogene-induced immortalization and growth-factor stimulation of naturally occurring central and peripheral nervous system stem cells. The nature of these cells and their suitability for transplantation into the CNS will be evaluated. PMID- 8239298 TI - Neurotrophic strategies for treating Alzheimer's disease: lessons from basic neurobiology and animal models. AB - Because neurotrophic factors can prevent natural and experimental cases of neural cell death and induce and maintain differentiation, they are especially attractive agents for the treatment of neurodegenerative diseases, such as Alzheimer's disease (AD). The present report argues for the specific role of particular families of trophic factors, such as neurotrophins (e.g., nerve growth factor [NGF]) and neurokines (e.g., ciliary neurotrophic factor [CNTF]), for the promotion of the survival and phenotype of subsets of central nervous system (CNS) neurons vulnerable in AD, such as basal forebrain cholinergic neurons and cortical projection neurons. Although there is ample evidence for the therapeutic role of NGF in experimental or natural injury of cholinergic neurons, not enough progress has been made on trophic models involving cortical neurons. Further understanding of the mechanisms of cell death in AD and elucidation of the transduction cascades of trophic factors will undoubtedly refine our current concepts of a neurotrophic treatment for AD. PMID- 8239299 TI - Selective signaling via unique M1 muscarinic agonists. AB - Rigid analogs of acetylcholine (ACh) were designed for selective actions at muscarinic receptor (mAChR) subtypes and distinct second messenger systems. AF102B, AF150, and AF151 are such rigid analogs of ACh. AF102B, AF150 and AF151 are centrally active M1 agonists. AF102B has a unique agonistic profile showing, inter alia: only part of the M1 electrophysiology of ACh and unusual binding parameters to mAChRs. AF150 and AF151 are more efficacious agonists than AF102B for M1 AChRS in rat cortex and in CHO cells stably transfected with the m1 AChR subtype. Notably, the selectivity of the new m1 agonists is reflected also by activation of select second messenger systems via distinct G-proteins. These compounds reflect a new pharmacological concept, tentatively defined as ligand selective signaling. Thus, agonist/m1AChR complexes may activate different combinations of signaling pathways, depending on the ligand used. Rigid agonists may activate a limited repertoire of signaling systems. In various animal models for Alzheimer's disease (AD) the agonists AF102B, AF150 and AF151, exhibited positive effects on mnemomic processes and a wide safety margin. Such agonists, and especially AF102B, can be considered as a rational treatment strategy for AD. PMID- 8239300 TI - Natural and synthetic Huperzine A: effect on cholinergic function in vitro and in vivo. AB - Huperzine A has been shown to be useful in the treatment of symptoms of dementia of the Alzheimer type. Our initial attempts to synthesize (-)Huperzine A resulted in the racemic mixture of (+/-)Huperzine A. We have therefore compared the in vitro and in vivo effects of (+/-)Huperzine A with those of (-)Huperzine A in rats. The results indicate a similar biological mechanism of action between the two, but that the racemic mixture of (+/-)Huperzine A has a weaker biological activity than the natural product (-)Huperzine A, presumably due to the presence in the mixture of (+)Huperzine A, which is considerably less potent than the ( )isomer. PMID- 8239301 TI - PKC translocation in rat brain cortex is promoted in vivo and in vitro by alpha glycerylphosphorylcholine, a cognition-enhancing drug. AB - Protein kinase C (PKC) activity was measured in soluble and particulate fractions of rat individual brain areas after treatment with alpha glycerylphosphorylcholine (GPC), a cognition-enhancing drug which promotes acetylcholine synthesis and release. The drug induced both in vivo and in vitro PKC translocation. In vivo, an increase of particulate PKC activity was observed 1 hour following the acute oral administration of a behaviorally active dose (600 mg/kg); the effect was transient. In vitro, GPC promoted PKC translocation in cortical slices at concentrations as low as 50 nM; the concentration-response curve was bell shaped. The increased PKC activity may be related to the cortical effects of GPC. PMID- 8239302 TI - Cholinergic neurotransmission in the hippocampus of aged rats: influence of L alpha-glycerylphosphorylcholine treatment. AB - The influence of aging and of L-alpha-glycerylphosphorylcholine (GFC) treatment on the acetylcholine synthesizing and degradating enzymes choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) and on cholinergic muscarinic M-1 and M-2 receptors were assessed in the hippocampus using immunocytochemical, histochemical and radioligand binding techniques, respectively. The investigation was performed on male Wistar rats of 2 months (young), 12 months (adult), and 27 months (old). Oral GFC was given at the dose of 100 mg/Kg/day from the 21st to the 27th month of age. ChAT revealed the highest immunostaining in the hippocampus of adult rats followed by young and old animals. The highest expression of AChE reactivity was noticeable in the hippocampus of adult rats followed by old and young animals. Treatment with GFC restored in part ChAT immunoreactivity and AChE reactivity in the hippocampus of aged rats. Muscarinic M-1 and M-2 receptors were labeled with [3H]-pirenzepine and [3H]-AF-DX-116 respectively. The density of M-1 muscarinic receptors decreased with age, whereas M-2 muscarinic receptors did not change. GFC treatment countered in part the loss of M-1 receptors in old rats and was without effect on M-2 receptors. PMID- 8239303 TI - Gangliosides and neurotrophic factors in neurodegenerative diseases: from experimental findings to clinical perspectives. AB - A large body of experimental data suggests that neurotrophic molecules and/or substances that facilitate their action could be pharmaceutical agents for neurodegenerative pathologies. In particular, it has been demonstrated that nerve growth factor (NGF) exerts a physiological role for forebrain cholinergic neurons, while brain-derived neurotrophic factor (BDNF) seems to play a relevant role in rescuing dopaminergic neurons following damage. In addition, gangliosides are reported to potentiate neurotrophic factor effects in vitro as well as in vivo. In this study we examined the effects of the monosialoganglioside GM1 in different experimental models. The responsiveness of forebrain cholinergic neurons following NGF +/- GM1 was evaluated by assessing choline acetyltransferase (ChAT) activity in hippocampus, septal area and striatum of behaviorally impaired 24-month-old rats. NGF was intracerebroventricularly (i.c.v.) infused for 2 weeks while GM1 was given systemically for 3 weeks, starting from the beginning of NGF infusion. Moreover, the possible protective effects of GM1 were assessed following exposure of cultured cerebellar granule cells and dopaminergic mesencephalic neurons to different doses of 6-OH-DOPA, a metabolite of the dopamine pathway which has excitotoxic properties and has been hypothesized to participate in the pathology of Parkinson's disease. GM1 treatment to aged rats was seen to potentiate the NGF-induced increase of ChAT activity in the striatum ipsilateral to the NGF infusion. Moreover, in the striatum contralateral to the NGF infusion, GM1 increased ChAT activity above the control values, whereas NGF treatment alone did not affect enzymatic activity. GM1 treatment of cerebellar granule cells and mesencephalic neurons counteracted the dose- and time-dependent neurotoxicity of 6-OH-DOPA. These data support the notion that GM1 might prove useful in treating those pathological conditions where trophic factor deficits and/or excitotoxin-related toxicity play an important role. PMID- 8239304 TI - Oral cytidine 5'-diphosphate choline administration to rats increases brain phospholipid levels. AB - Exogenous cytidine 5'diphosphocholine (CDP-choline) is completely metabolized to circulating cytidine and choline. Both compounds enter the brain and can be used in phosphatidylcholine (PC) synthesis via the Kennedy (CDP-choline) cycle. We administered oral CDP-choline to 12 month-old rats (500 mg/kg/day) for 21, 42, or 90 days to determine whether this treatment would alter brain levels of PC and the other structural phospholipids, phosphatidylserine (PS) and phosphatidylethanolamine (PE). After 42 days, brain PC levels increased significantly (p < 0.01) by 23.3%; after 90 days PC increased by 30% (p < 0.01), PS by 37.2% (p < 0.01), and PE by 13% (not significant). The ratios of each of the phospholipids to total membrane phospholipids were unchanged. These data demonstrate that repeated oral CDP-choline administration can increase the amounts of phospholipids in brain membranes, thus providing a rationale for using this compound in brain diseases that damage neurons. PMID- 8239305 TI - Effect of CDP-choline on cognition and immune function in Alzheimer's disease and multi-infarct dementia. AB - The cholinergic dysfunction present in Alzheimer's disease (AD) might be due to a specific vulnerability of cholinergic neurons linked to neurotrophic imbalance, neuroimmune impairment, and/or direct effects of beta-amyloid deposition and NFT formation in ACh neurons. The presence of abnormal epitopes exposed on neuronal membranes may contribute to the activation of resting microglia initiating a neuroimmune cascade leading to cell destruction. According to this hypothesis, a multifactorial treatment in AD should produce: 1) inhibition of beta-amyloid and NFT formation; 2) restoration of neuronal membrane integrity; and 3) control of neuroimmune auto-aggression. Since interleukin-1 (IL-1) is an APP gene promoter showing a progressive increase in body fluids in parallel with mental deterioration in AD patients, we have studied the effects of CDP-choline on cognition, several biological parameters, and IL-1 beta production in AD and multi-infarct dementia (MID) in order to elucidate whether this compound alone or in combination with other drugs is able to restore immune function and improve mental performance in senile dementia. PMID- 8239306 TI - Acetyl-L-carnitine and Alzheimer's disease: pharmacological considerations beyond the cholinergic sphere. AB - Since ALCAR and L-carnitine are "shuttles" of long chain fatty acids between the cytosol and the mitochondria to undergo beta-oxidation, they play an essential role in energy production and in clearing toxic accumulations of fatty acids in the mitochondria. ALCAR has been considered of potential use in senile dementia of the Alzheimer type (SDAT) because of its ability to serve as a precursor for acetylcholine. However, pharmacological studies with ALCAR in animals have demonstrated its facility to maximize energy production and promote cellular membrane stability, particularly its ability to restore membranal changes that are age-related. Since recent investigations have implicated abnormal energy processing leading to cell death, and severity-dependent membrane disruption in the pathology of Alzheimer's disease, we speculate that the beneficial effects associated with ALCAR administration in Alzheimer patients are due not only to its cholinergic properties, but also to its ability to support physiological cellular functioning at the mitochondrial level. This hypothetical mechanism of action is discussed with respect to compelling supportive animal studies and recent observations of significant decrease of carnitine acetyltransferase (the catalyst of L-carnitine acylation to acetyl-L-carnitine) in autopsied Alzheimer brains. PMID- 8239307 TI - Activation PET as an instrument to determine therapeutic efficacy in Alzheimer's disease. AB - Forty patients with probable Alzheimer's disease (AD) were selected from a pool of 80 patients and assigned to 4 groups. Each received either social support, cognitive training only, or cognitive training in combination with pyritinol or phosphatidylserine. Treatment duration was 6 months. Before and after treatment the patients underwent neuropsychological testing as well as measurement of the regional cerebral metabolic rate for glucose using positron emission tomography (PET) and 2[18F]-fluoro-2-deoxy-D-glucose (FDG). Before treatment, the groups were comparable in respect to resting and activated glucose pattern achieved by a visual recognition task. They did not differ in scores of a neuropsychological test battery. After the treatment period the group with cognitive training + phosphatidylserine showed a significant glucose enhancement during the stimulation tasks in various brain regions, and an improvement in cognitive functioning compared to the other groups. The group with cognitive training + pyritinol had better stimulation effect as that of the social support group indicating that a combination of cognitive training + pharmacological intervention was superior than that of cognitive training alone. PMID- 8239308 TI - The use of the Computerized Neuropsychological Test Battery (CNTB) in an efficacy and safety trial of BMY 21,502 in Alzheimer's disease. AB - BMY 21,502 is a nootropic which protects memory and enhances long-term potentiation according to preclinical findings. Alzheimer's disease (AD) patients who were diagnosed by DSM-III-R and NINCDS-ADRDA criteria were enrolled in a 12 week double-blind investigation of BMY 21,502 vs. placebo at 300 mg tid. The study design included a 1-week placebo lead-in and a 4-week placebo washout in addition to the 12-week double-blind treatment period. Efficacy was assessed with the Alzheimer's Disease Assessment Scale (ADAS) and the Computerized Neuropsychological Test Battery (CNTB) at weeks 4, 8, 12, and 16. Clinical Global Impression (CGI) assessments were also performed biweekly. Sixty-nine patients (28M, 41F; mean age 72 years, range 54 to 92 years) were enrolled in the study. Baseline Mini-Mental Status Examination (MMSE) scores ranged from 16 to 26 (mean 23.5) in patients on active drug (n = 34), and from 15 to 26 (mean 22.5) in placebo patients (n = 35). Baseline efficacy scores were comparable for drug and placebo patients (p > 0.05). Twelve (35%) patients who received BMY 21,502 withdrew from the study, 8 (24%) due to adverse events. Three (9%) patients who received placebo withdrew from the study, all due to adverse events. Patients on active drug who were valid for analysis of efficacy (n = 22) showed a mean decrease in ADAS of -1.5 at week 12, vs. a mean change of -0.5 in patients who received placebo (n = 32), although there was no significant difference between the two (p > 0.05). Correlations between the CNTB summary scores and ADAS cognitive subscores were, nevertheless, highly significant at baseline (r = 0.83, p = 0.0001) and week 12 (r = -0.83, p = 0.0001). Correlations between the word list learning, spatial, and naming subtests of the ADAS and CNTB were also highly significant (p = 0.0001). Although modest, the findings for active drug vs. placebo response in this study suggest that BMY 21,502 should be further investigated, with a larger study population, in order to fully determine the compound's potential efficacy. PMID- 8239309 TI - Hyperactivation of signal transduction systems in Alzheimer's disease. AB - A compromise or deregulation in signal transduction cascades could adversely affect cellular functions and possibly contribute to cell death. In recent years, it has become increasingly apparent that pronounced activation of neuronal signal transduction systems is a characteristic of AD brain. There is evidence that signal transduction systems play a role in the formation or development of these pathological features of AD. Aberrant activity and localization of components of signaling mechanisms (growth factors, their receptors, protein kinases, phosphoprotein phosphatases, and phosphoproteins) are closely associated with the intracellular accumulation of PHF, the extracellular deposition of amyloid, and the formation of neuritic plaques in AD brain. In particular, immunohistochemical studies reveal increased levels of neuronal staining for APP, possibly an important growth factor in AD, both in frontal cortex and hippocampus. Anti-APP immunostaining is also associated with the neuritic component of plaques. Additionally, PKC(beta II) immunostaining is increased in the neuronal cell body and neuropil of AD samples, particularly in association with plaques, suggesting a postsynaptic involvement of this enzyme. On the other hand, PKC(beta I) immunostaining is associated with axonal staining particularly in the sprouting neurites of plaques. Sprouting neuritic components of plaques are immunopositive with other growth-associated proteins, such as GAP43, MARCKS, and spectrin. Immunoreactivity of other members of signal transduction systems such as Fos and stathmin are all increased in AD hippocampal neurons. On the other hand, several protein kinases and phosphoproteins were immunolocalized to tangles. Thus, the hyperactivation and dysfunction of signal transduction systems could be involved in the pathogenesis of AD. PMID- 8239310 TI - Changes in G protein-mediated signal transduction in aging and Alzheimer's disease. AB - Previous reports have shown that there are age-related reductions in muscarinic receptor (mAChR) sensitivity to agonist stimulation. Our research has elucidated the mechanisms involved in this loss. These studies have shown that this decline is the result of decreases in mAChR concentration, reductions in the number of neuronal cells, and altered phosphoinositide (PI)-mediated signal transduction (ST). The decrements in PI-mediated ST are observed as a reduced ability of muscarinic (m) agonists to enhance K(+)-evoked release of DA (K+ ERDA) from striatal slices from old rats. Additional experiments indicated that the locus of the ST deficits appears to be at the mAChR-G protein interface, since attempts to bypass this interface reduced m-enhanced K+ ERDA deficits in the striata from old rats. Moreover, it appears that the ability of mAChR to decouple from their respective G proteins is reduced as a function of age, since carbachol-stimulated low KM GTPase activity was found to be reduced in hippocampal and striatal tissue obtained from old rats. Similar findings were observed in this parameter in AD hippocampus and basal ganglia. Further reductions were seen in carbachol stimulated low KM GTPase as a function of the duration of the disease. Results are discussed in terms of structural membrane alterations in aging and disease that may lead to reductions in the efficacy of receptor-G protein coupling/uncoupling. PMID- 8239311 TI - Aberrant phosphoinositide metabolism in Alzheimer's disease. AB - Since phosphoinositide-specific phospholipase C (PLC) is one of the key molecules in signal transduction, its involvement was assessed in Alzheimer's disease (AD). The phosphatidyl-inositol (PI)-specific PLC activity in the Alzheimer cytosolic and particulate fractions was not significantly different from that in the control fractions. The PI-specific PLC activity as a function of the free Ca2+ concentration was also similar between control and Alzheimer brains. These results suggest that the PI-specific PLC activity is not altered in AD. Immunostaining of a specific antibody against the PLC isozyme, PLC-delta, demonstrated that this enzyme was abnormally accumulated in neurofibrillary tangles (NFT), the neurites surrounding senile plaque (SP) cores, and neuropil threads in AD brains. Western blot analysis confirmed that PLC-delta was concentrated in the paired helical filament (PHF)-rich fraction of AD brains. PLC delta marked the same neurons containing tau immunoreactivity and yet tau and PLC delta often marked different structures within the same neuron, with tau more clearly on NFT and PLC-delta covering it superficially. The double stain with PLC delta and basic fibroblast growth factor (bFGF) binding suggest that PLC-delta is an intracellular marker, showing little overlap with bFGF binding, an extracellular marker. All of this was consistent with the electron microscopy, with PLC-delta being NFT associated. Antibodies to other PLC isozymes did not produce positive immunostaining of these pathologic structures. Moreover, diffuse and amorphous deposits of PLC-delta were found to precede the accumulation of fibrillary deposits. These results suggest that PLC-delta accumulation plays a possible role in the formation of intraneuronal inclusions in AD. PMID- 8239312 TI - NMDA receptor status in elderly normal individuals and those with Alzheimer's disease. AB - The status of NMDA receptors in the brains of normal aged individuals and those with Alzheimer's disease was investigated. The binding of [3H]3-((+-)-2 carboxypiperazin-4-yl)propyl phosphonic acid ([3H]CPP) to NMDA antagonist preferring sites on frontal and temporal cortical synaptic membranes was assessed. Binding could be resolved into two components, one of high and the other of low affinity. Pharmacologically, the two sites were qualitatively similar. Considerable intersubject variation in binding parameters was detected, but no significant differences were found between the mean values for the control and Alzheimer's disease groups. This study indicates that, when changes in receptor integrity occur in individual patients, these may be occluded because of the large variations between individuals. PMID- 8239313 TI - X-ray diffraction analysis of brain lipid membrane structure in Alzheimer's disease and beta-amyloid peptide interactions. AB - Small angle x-ray diffraction analysis of Alzheimer's disease (AD) lipid membranes reconstituted from cortical gray matter showed significant, reproducible structure changes relative to age-matched control samples. Specifically, there was an average 4 A reduction in the lipid bilayer width and marked changes in membrane electron density profiles of AD cortical samples. There were no significant structure differences in the membrane bilayers isolated from an unaffected region (cerebellum) of the AD brain. Lipid and protein analysis of six AD and six age-matched controls showed that the phospholipid:protein mass ratio was unchanged, but that the unesterified cholesterol:phospholipid (C:PL) mole ratio decreased by 30% in the AD temporal gyrus relative to age-matched controls. The C:PL mole ratio was not significantly different for samples prepared from cerebellum of AD versus control patients. X ray diffraction analysis of a cholesterol-enriched AD sample demonstrated a virtual restoration of the normal membrane bilayer width and electron density profile, suggesting that the cholesterol deficit played a major role in the AD lipid membrane structure perturbation. Addition of beta-amyloid peptide to bovine brain phospholipid membranes significantly changed the electron density associated with the hydrocarbon core. Alterations in the composition and structure of the membrane bilayer may play an important role in the pathophysiology of AD by altering the activity and catabolism of membrane-bound proteins, including the beta-amyloid precursor protein. PMID- 8239314 TI - Synaptic pathology of Alzheimer's disease. AB - Prospective clinico-pathological studies on dementia in Alzheimer's disease (AD), performed during the past decades, revealed a relatively poor correlation between the degree of clinical deficit and the severity of the typical neuropathological lesions of AD, the amyloid plaques and the neurofibrillary tangles. More recent data, obtained by electron microscopy, immunocytochemical as well as immunochemical techniques indicate that synaptic loss may be a better structural correlate of dementia than other brain lesions. Synaptic pathology is reflected by a loss of all major components of small synaptic vesicles and most peptides, stored in large dense cored vesicles. The significant increase of chromogranin A proprotein, a major component of large dense cored vesicles, may rather represent a defect of protein processing than preservation of a specific synaptic subpopulation. Within the brain of AD patients, the degree of synaptic loss is uneven. Most prominent reduction of synapses is found in the outer parts of the dentate gyrus molecular layer, possibly reflecting the destruction of neurons, located in the layer 2 of the entorhinal cortex. However, within the neocortex, no preferential loss of synapses in any of the cortical layers has been found. Cerebral amyloid deposition in diffuse plaques has little effect on synapse density and structure. However, within the dense amyloid core of a classical plaque, synapses are completely lost. In the surrounding neuritic portion of the plaques, synaptophysin reactivity is frequently increased, due to enlargement of synaptic boutons and to accumulation of synaptophysin in dystrophic axons. Although the reason for synapse loss in AD is yet unknown, most results suggest that it may reflect degeneration of neurons, projecting into the respective cortical areas. PMID- 8239315 TI - Protease inhibitors and indolamines selectively inhibit cholinesterases in the histopathologic structures of Alzheimer's disease. AB - Neurofibrillary tangles and amyloid plaques express acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity in Alzheimer's disease. We had found that traditional AChE inhibitors such as BW284C51, tacrine and physostigmine were more potent inhibitors of the AChE in normal axons and cell bodies than of the AChE in plaques and tangles. We now report that the reverse pattern is seen with indolamines, 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 neurons and axons. This shift in enzymatic affinity may lead to the abnormal protein processing which is thought to play a major role in the pathogenesis of AD. The availability of pharmacological and dietary means for altering brain indolamines raises novel therapeutic possibilities for inhibiting the abnormal cholinesterase activity associated with Alzheimer's disease. PMID- 8239316 TI - Free radicals in the genesis of Alzheimer's disease. AB - As part of an ongoing investigation of the role of oxygen free radicals in Alzheimer's disease (AD), the formation of peroxidation products, the activities of free radical defense enzymes, and the level of total iron were determined in autopsy brain tissue from donors with AD and from age-matched non-demented donors. Calcium uptake was also investigated in mitochondria harvested from fibroblasts grown in tissue culture from skin samples taken from brain donors. Compared to controls, homogenates of AD frontal cortex produced elevated levels of peroxidation products and this difference was amplified in a dose-dependent manner by iron (1 to 200 microM). Peroxidation produced by 200 microM iron was reduced dose dependently by the lazaroid U-74500A. The IC50 was 10 microM in AD cortex and 2.5 microM in controls. Superoxide dismutase (SOD), one of the free radical defensive enzymes, was reduced by 25 to 35% in AD frontal cortex, hippocampus and cerebellum. In other brain areas, SOD did not differ between AD and control. The activities of catalase and glutathione peroxidase were the same in AD and control samples. Endogenous iron levels were higher in AD frontal cortex (2.5 nmol/mg protein) than in controls (1.5 nmol/mg protein). Calcium uptake by AD fibroblast mitochondria is 50% lower than in controls under basal conditions. Following exposure to 200 microM iron, mitochondrial calcium uptake is increased by 58% in AD and by 38% in controls. Pretreatment with 200 microM U 74500A or 1 mM deferoxamine, prior to exposure to 200 microM iron, gave complete protection to control mitochondria but gave only partial protection to AD mitochondria. These studies indicate that in AD, both CNS and peripheral cells show increased sensitivity to oxygen free radicals. The source of this increased sensitivity has not yet been identified but could reflect either reduced free radical defenses or increased free radical formation or both. Work is underway using electron paramagnetic resonance spectrometry to determine in vivo, premortem free radical activity in AD patients. PMID- 8239317 TI - Abnormalities in brain glucose utilization and its impact on cellular and molecular mechanisms in sporadic dementia of Alzheimer type. AB - Brain glucose utilization and ATP formation were found to be reduced to 54% and 81%, respectively, of control values in incipient sporadic dementia of Alzheimer type, causing reduced availability of the glucose-derived neurotransmitter acetylcholine. With respect to energy shortage, impacts on energy-dependent processes such as synaptic transmission, ion homeostasis, protein processing and degradation, extracellular transmission, and extracellular phosphorylation may be expected. Normal processing of the amyloid precursor protein was demonstrated to occur via a muscarinergic acetylcholine M1 and M3 receptor-mediated signal transduction pathway. Since both the muscarinergic acetylcholine receptors on pyramidal neurons and G proteins were found to be unaltered in DAT, the possibility is discussed that the diminution of glucose utilization and the energy shortage in DAT brain may contribute considerably to abnormal amyloid precursor protein processing and thus to secondary amyloid formation. PMID- 8239318 TI - Dopamine D2 receptors in normal human brain: effect of age measured by positron emission tomography (PET) and [11C]-raclopride. AB - Human post-mortem and animal experimental results suggest a decline of the cerebral dopaminergic neuronal system with age. In this study, the radiotracer [11C]raclopride (dopamine D2 antagonist) and positron emission tomography were applied to determine the effect of age on striatal dopamine D2 receptors in 32 healthy volunteer subjects (age range 21-68 years). Subjects were divided in two age groups on the basis of median age (31 years). An index for specific tracer uptake was calculated for caudate nucleus and putamen. Uptake indices in the older group of subjects were reduced on average 26% in putamen and 20% in caudate nucleus. The decline appeared to be steep until 30 years, but slower afterwards. After 30 years of age the decline of specific raclopride binding was found to be 0.6% per year. These results suggest that dopamine D2 receptor binding sites (mainly post-synaptically located) decrease as a consequence of normal aging in parallel with the decline of the pre-synaptic nigrostriatal dopaminergic system. PMID- 8239319 TI - Comparative effects of aging process on phosphatidylcholine biosynthesis pathway: a key role for CTP-phosphocholine cytidylyltransferase? AB - The effects of aging on phosphatidylcholine (PtdCho) biosynthesis were investigated in liver and brain subcellular fractions of the rat, by studying the activity and regulation of CTP phosphocholine cytidylyltransferase (CT), the rate limiting enzyme in PtdCho biosyntheses. With both tissues, CT activity was present in cytosolic and microsomal fractions, but in brain, CT activity seemed to escape to an inhibiting feed back mechanism. In brain, CT activity was greater in the microsomal fraction, whilst in liver, a higher CT activity was seen in the cytosolic fraction. In liver fractions of aged animals, there was no significant change in CT activity or its sensitivity to negative feedback regulation, as compared to young animals. In contrast, a progressive age related decline in CT activity was observed in the brain microsomal fraction. Furthermore, the incorporation of newly formed CDPCho into PtdCho was also reduced in aged animals, and paralleled the decreased incorporation of choline in PtdCho. The age related decrease in CT activity cannot be explained by product feed back inhibition or decreased diacylglycerol levels. Since PtdCho is a major membrane lipid, the reduction in CT activity may lead to a decreased membrane integrity and fluidity during the aging process, and these effects may be greater in neuronal cells. PMID- 8239320 TI - Regulation and expression of the Alzheimer's beta/A4 amyloid protein precursor in health, disease, and Down's syndrome. AB - A four- to fivefold overexpression of the gene for the Alzheimer beta/A4 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 (AD) pathology in DS compared to the normal population. It is therefore likely that a deregulated overexpression of the APP gene is a risk factor for the beta/A4 amyloid formation. To test this hypothesis and to get a better understanding of how APP expression is regulated, we studied the 5' control region of the human APP gene, alternative splicing of the 19 APP exons, and APP biogenesis, metabolism and function. The analysis of the APP promoter revealed its similarity with those of housekeeping genes by the presence of a GC-rich region around the transcription start site and the lack of a TATA box. Gene transfer experiments showed this GC-rich region to contain overlapping binding sites for different transcription factors whose binding is mutually excluded. An imbalance between these factors may cause APP overexpression and predispose to AD pathology. Another putative risk factor for AD is regulation of splicing of exon 7 in APP mRNA's which changes in brain during aging. This is relevant for APP processing since exon 7 codes for a Kunitz protease inhibitory domain. Investigation of further splicing adjacent to the beta/A4 exons 16 and 17 which might also interfere with APP processing led to the identification of the leukocyte-derived (L-APP) splice forms which lack exon 15. In brain this splicing occurs in activated astrocytes and microglia. The localization of APP at synaptic sites in brain suggests that APP regulation and expression are critical determinants of a potential and early impairment of central synapses. This may be the case during pathological evolution of AD and DS when beta/A4 derived from synaptic APP is converted to beta/A4 amyloid by radical generation. PMID- 8239321 TI - Angiotensin-converting enzyme inhibitor treatment for young normotensive diabetic subjects: a two-year trial. AB - Microangiopathy characterizes both diabetic retinopathy and nephropathy. It is currently unclear which diabetic subjects should be treated with angiotensin converting enzyme (ACE) inhibitors. A double-blind, placebo-controlled protocol was implemented using captopril to treat subjects with Type I diabetes, early diabetic nephropathy (albumin excretion rates, 20-200 micrograms/min), and normal blood pressures. After two years, the final eye grades were improved in two treated subjects but not in any of the controls. Three control and one treated subject showed worsening of their eye grade after two years (P < .001, by chi square test). Significant differences in renal albumin excretion were not seen between the two groups. The distribution of changes in retinal grades in the treatment group compared with the placebo group was improved after two years. Studies of larger numbers of patients will be necessary to determine if ACE inhibitors should be used routinely in subjects with diabetic retinopathy and to determine which subjects are most likely to respond. PMID- 8239322 TI - Handling punctate keratitis resulting from systemic cytarabine. AB - A young patient with acute myelogenous leukemia was treated with high-dose cytarabine (3.0 g/m2 every 12 hours). After five days of treatment, he developed severe photophobia and decreased visual acuity. On examination, progressive punctate keratitis was found. We discuss the mechanism of the cytarabine-induced corneal toxicity and the prophylactic use of topical corticosteroids. PMID- 8239323 TI - Successful strabismus surgery in a child with Moebius syndrome. AB - Moebius syndrome is characterized by congenital paralysis, usually bilateral, of the lateral rectus muscles and the muscles of facial expression. Esotropia is common in these patients. Few reports exist of the surgical results after treatment of strabismus in such patients. We report the case of a 19-month-old white boy with Moebius syndrome who demonstrated an A-pattern esotropia with orthotropia in down-gaze. Use of a conventional surgical approach for an A pattern esotropia and a bilateral medial rectus recession with a one-half tendon width supraplacement produced the desired result of orthotropia in primary gaze in our patient. PMID- 8239324 TI - Combined cataract and strabismus surgery. AB - A patient with cataracts and congenital exotropia underwent combined cataract and strabismus surgery OU. A lateral rectus recession plus an extracapsular cataract extraction with intraocular lens implantation was done OD first; three months later, this procedure was repeated OS. The patient's postoperative course was benign in both cases, and her strabismus resolved after the second operation. A combined surgical approach to cataracts and strabismus (where only a single muscle is involved) was safe and useful in restoring this patient's vision, binocularity, and appearance. PMID- 8239325 TI - Myopic astigmatism and simple myopia correction. AB - A new technique for the treatment of myopic astigmatism and simple myopia correction is described. The technique is based on bioengineering concepts previously studied by the authors that accurately and mathematically describe the transformation of an ellipsoidal surface (astigmatic eye) into a spheroidal one (nonastigmatic eye) by means of minimal cuts (keratotomies). The surgical procedure is described using selected cases to illustrate the alternatives the model can handle. The model itself is not presented because the calculations were done by computer. PMID- 8239326 TI - Pseudotumor cerebri induced by vitamin A combined with minocycline. AB - A 16-year-old girl complained about a headache of one-month's duration, accompanied by vertical diplopia that had appeared ten days earlier. The girl reported receiving vitamin A and minocycline to treat acne vulgaris for the previous six weeks. An examination revealed bilateral optic disc edema. Normal computed tomographic and magnetic resonance imaging examinations enabled a diagnosis of pseudotumor cerebri to be made. Soon after discontinuation of those medications, the headaches and diplopia diminished. We suggest a periodic ophthalmologic examination during systemic therapy with vitamin A combined with minocycline to detect the early occurrence of pseudotumor cerebri. PMID- 8239327 TI - Endophthalmitis caused by Enterobacter cloacae. AB - Cataract extraction with placement of intraocular lenses is the most common intraocular procedure done today, with endophthalmitis as its most devastating complication. To our knowledge, this is only the third reported case of postoperative endophthalmitis caused by the gram-negative bacillus, Enterobacter cloacae. The course of infection, the treatment, and the laboratory identification of this organism are presented. It is important to add this proven case of gram-negative endophthalmitis to our growing knowledge of causes of this disease because, despite aggressive treatment with vitrectomy and intravitreal antibiotics, the infection was so virulent that it led to eventual phthisis bulbi. PMID- 8239328 TI - Chest x-rays before ophthalmic surgery. AB - We retrospectively reviewed clinical records and preoperative chest x-ray films of 636 patients who had been admitted for ophthalmic surgery in 1990 and 1991. In our hospital, a preoperative chest roentgenogram routinely was examined for each patient. A total of 228 patients (35.8%) had an abnormal preoperative result. These abnormalities were more frequent in patients older than 40 years of age than in younger patients. Pulmonary fibrosis and calcification were the most common abnormal findings. Also, we noted that 202 patients (31.8%) had a chronic, previously known, or suspected, finding, and 26 patients (4.1%) had a new or unsuspected finding. The surgery was postponed or canceled in five patients (0.8%) because of the abnormal x-ray results. PMID- 8239329 TI - Congenital cataract in the right eye and primary clinical anophthalmos of the left eye in a patient with cerebellar hypoplasia. AB - We examined a 26-year-old man who had an opaque lens OD and clinical anophthalmos OS. Low densities in the left cerebellar hemisphere and vermis and dilated ventricles were noted on a computed tomographic scan of his brain. We believe that the association of developmental cataract OD, clinical anophthalmos OS, and cerebellar hypoplasia in our patient may be rare. PMID- 8239330 TI - Recent advances in lasers in Otolaryngology. AB - Laser surgery has been an integral part of otolaryngology procedures since first applied to the larynx nearly 20 years ago. New wavelengths and delivery systems are now available that may prove beneficial but also offer stimulating areas of research. Safe and judicious use of these devices is essential. This editorial describes current future applications of lasers in our specialty. PMID- 8239331 TI - [Abnormal movements of the larynx. Diagnostic approach and therapeutic perspectives]. AB - Laryngeal movement disorders of neurological cause are actually misunderstood. In the course of chorea, or tardive post neuroleptic dyskinesia, vocal, swallowing for threatening breathing troubles could occur. Ten patients with generalized dyskinesias were studied by endoscopy with flexible laryngoscope and laryngeal electromyogram. Four of them were choreic and six had tardive dyskinesia (one had a severe breathing disorder). Nine patients had abnormal movement disorders (MD) involving intrinsic laryngeal musculature. MD were spontaneous or were triggered by vocalization. These facts suggest that dyskinesia as they involved the upper airway tract could be life threatening. Laryngeal electromyography is a useful method for diagnosis and treatment of these dyskinesia. As in spasmodic dysphonia, botulinum toxin could be helpful in this local treatment. PMID- 8239332 TI - [Conservative surgery of the larynx as an alternative to total laryngectomy after neoadjuvant chemotherapy. Initial results]. AB - A retrospective study of 149 squamous cell carcinomas of the larynx and epilarynx in whom a total laryngectomy was initially planned is reported. All the patients were treated by neoadjuvant chemotherapy. After completion of chemotherapy, the initially planned therapeutic protocol was modified in 56 patients (37.6%) in order to preserve the laryngeal physiologic functions. A total laryngectomy was performed in 23 patients (41%), radiation therapy was administered to 14 (25%) patients chemotherapy was prolonged in 15 patients (26.8%) and four deaths were related to chemotherapy toxicity (7.2%). Actuarial survival of patients treated by partial laryngectomy was statistically higher than survivals of patients treated either by radiation therapy or prolonged chemotherapy. After completion of neoadjuvant therapy patients who refused a total laryngectomy had the worst survival of the series. These results suggest that surgery of the residual tumor (partial laryngectomy) after neoadjuvant chemotherapy could be indicated for selected laryngeal carcinoma in whom a total laryngectomy was initially planned. PMID- 8239333 TI - [Correlations between anatomy and x-ray computed tomography in cancers of the pharyngo-larynx. Apropos of a prospective study of 35 patients]. AB - A prospective study has been conducted in 35 cases of pharyngolaryngeal's carcinoma, between december 1990 and june 1992 in the department of Head and Neck Surgery of Strasbourg. The authors compared, in a statistic analysis, Computed Tomography and histopathologic's correlations. The operative specimen were sectioned in the horizontal plane to correspond with the CT sections. 11 structures were studied more particularly. Results revealed an excellent sensibility for the study of the pre-epiglottic spaces (100%). Inversely, the study of the para glottic space is less good (69% of sensibility). We found that the CT Scan was of no value for the study of the thyroid cartilage, particularly for detection of subclinical lesions. Overall, this study allowed to confirm the value of the CT Scan in the investigation of pharyngolaryngeal tumors but its limits also. PMID- 8239334 TI - [Spectral study of laryngotracheal sounds. Methodological approach and perspectives]. AB - Prior studies have shown that laryngotracheal sounds are vectors of objective information which indicate the origins and mechanisms of sound production. The first objective of this study is to confirm these earlier findings on a larger scale. The second objective is to develop a simple apparatus which permits the rapid acquisition and analysis of information for a prompt diagnosis. This study will be carried out in infants referred to the Department of otolaryngology of the Kremblin-Bicetre Hospital over the next two years. The material and methods used are described. PMID- 8239335 TI - [Results of the surgical treatment of cancers of the supraglottic area]. AB - The authors report a retrospective analysis of the results of surgery in 169 patients with supraglottic cancer referred to the Lille ENT clinic from 1976 to 1985. Partial surgery was performed in 53% of patients versus total laryngectomy in 47%. The 5 year survival rates in the patient groups treated by supraglottic laryngectomy and supracricoid laryngectomy with cricohyoidopexy were similar, 69.4% and 75.2%, respectively. The 5 year survival rate in the patient group treated by total laryngectomy or by total laryngectomy extended to the base of tongue was 62.8%. PMID- 8239336 TI - [Reoperation of tumors of the parotid gland. Technical approach and consequences for the 7th cranial nerve. Apropos of 22 cases]. AB - Second or more surgical procedures on parotid are usually difficult and may induce injury on the facial nerve. The authors report their experience about 42 patients. The choice between a total parotidectomy or a surgical excision of a tumor depends of the number of surgical procedure, the type of initial procedure and the histological type of the tumor. Second surgical procedures for a wrong initial histological diagnosis (11 cases) were always a total parotidectomy with facial nerve preservation. The risk of a partial or total facial palsy is higher after several recurrences (3 of 8 cases). The facial nerve had to be resected in 3 of 9 cases of malignant tumors recurrences. The initial treatment of a tumor of the parotid must be radical so it can prevent for further surgical procedures and then avoid a facial nerve injury. PMID- 8239337 TI - [Value of the association of normovolemic dilution and hyperbaric oxygenation in the treatment of sudden deafness. A retrospective study]. AB - The multiple treatments of sudden deafness shows how this pathology still remains quite unknown. The authors present a retrospective study of 87 patients treated by normovolemic hemodilution associated to hyperbaric oxygenation. They obtain a total a 60% of significant recovery (ratio between hearing gain and initial hearing loss, above 25%) and in severe hearing loss (threshold between 70 and 90 dB) 60% of good results (ratio above 50%). The importance of the initial form of audiogram and the presence of dizziness as prognostic factors is not confirmed. On the other hand, the evolution of tinnitus is correlated with the deafness and it is a supplementary means to evaluate the therapeutic efficiency. Moreover the persistence of tinnitus represents an important after effect. Sudden deafness still remains a medical emergency and the delay for carrying out any treatment should be as short as possible. On the other hand it is possible to reduce hospital stay by two sessions of hyperbaric oxygenation per day. PMID- 8239338 TI - [Oral manifestations of zona. Apropos of a case]. AB - Oral manifestations of herpes zoster are very less common than cutaneous. Only a few cases of oral herpes zoster in children had been already described. Authors report a case of maxillary superior nerve's herpes zoster. Oral lesions are encountered in case of viral disease of the second and third branch of trigeminal nerve (VII, VIII). Dental pain is usually the first sign and can induce misdiagnosis. The diagnosis is based on the specific ulcerative lesions, strictly unilateral, developed in the field of sensitive innervation of the maxillary nerves. The use of antiviral drugs seems to be actually the best treatment. PMID- 8239339 TI - [Medical innovation. Patents and confidentiality]. AB - Enhanced knowledge of medicine and commercialization of an innovation (product, equipment, drug ...) are the classical aims of all medical research. Only the successful commercialization of the innovation allows the largest number of patients to benefit from the achievements of this research. Successful commercialization depends on the effective cooperation between the medical researcher who possess the potential for innovation and industry which provides the means necessary for its development. This cooperation is facilitated if the innovation is patented and if researcher ans industry work together under a contract of confidentiality. In order to improve the cooperation necessary between industry, research scientists, and clinicians, we review the current rules and regulations pertaining to patents and confidentiality. An annex is included with examples of a consultants contract and a secrecy agreement. PMID- 8239340 TI - [How to alleviate the effect of audiometry in practice]. PMID- 8239341 TI - Repair of a laryngotracheoesophageal cleft in an infant by means of extracorporeal membrane oxygenation. AB - Few survivors have been reported following attempted repair of laryngotracheoesophageal clefts (LTECs). The major challenge is maintaining oxygenation, both during the surgical repair and during the postoperative period of healing. We report a neonate with an LTEC extending to the carina whose successful repair was facilitated by extracorporeal membrane oxygenation (ECMO) begun intraoperatively and continued postoperatively for 11 days. The intraoperative surgical exposure of the defect was excellent. Postoperative trauma to the fresh tracheal repair from ventilatory pressures and endotracheal tube motion was eliminated through the use of ECMO. The patient was discharged without a tracheotomy and with a normal voice, cry, and swallow. According to this result, the use of ECMO may represent a significant advance in facilitating the correction of major laryngotracheoesophageal anomalies. The rationale, advantages, disadvantages, and potential pitfalls of this approach are presented, as well as preoperative and postoperative documentation of our results. PMID- 8239342 TI - Tracheoscope: an old instrument with new applications. AB - The tracheoscope is a bronchoscope without the side ventilating ports at the distal tip. It has been used for decades, implementing diagnostic and therapeutic procedures in the subglottis and cervical and thoracic trachea. The closed tube tracheoscope allows excellent control of the airway with maintenance of continuous anesthesia and ventilation. Flexible fiber-directed lasers, such as the potassium-titanyl phosphate laser, can be used with the tracheoscope for laser resection or vaporization of diseased tissue from just below the vocal cords to any level within the trachea. Controlled anesthesia and smoke evacuation are maintained throughout the operation, allowing a relatively safe endoscopic procedure. Disorders such as subglottic hemangioma, subglottic or tracheal webs, papillomatosis, granuloma, and a variety of neoplastic growths can be excised, debulked, or submitted to biopsy by means of this instrument. In the neonatal and infant airway, use of the tracheoscope concomitantly with rigid telescopes has been particularly rewarding. PMID- 8239343 TI - Phrenic nerve reinnervation of the cat's larynx: a new technique with proven success. AB - Reinnervation of the posterior cricoarytenoid muscle (PCA) should provide vocal cord abduction on inspiration, and passive adduction to enable phonation. Previous investigators have shown that reinnervation is possible, but results have not been clinically encouraging. When reinnervation was successful, the question remained whether it was provided by the transplanted nerve or by the ingrowth of adjacent nerves. In this study the phrenic nerve was transplanted directly into the PCA in a series of 12 cats. Fibrin glue was used to overcome nerve trauma and to prevent retraction of the nerve from the PCA. Laryngoscopy, electromyography, and retrograde labeling of the phrenic motoneurons provided evidence of functional reinnervation in 9 cats. Partial or complete failure in the remaining 3 was due to retraction of the nerve from the muscle. These results appear to justify trials of the procedure in humans. PMID- 8239344 TI - Hydroxylapatite laryngeal implants for medialization. Preliminary report. AB - Laryngeal implantation for medialization has improved our ability to manage the patient with vocal fold motion impairment. We present preliminary data evaluating the use of preformed hydroxylapatite laryngeal implants and instrumentation for rapid determination of implant size and position. A window in the thyroid ala is created by means of a standard fenestra template. One of 5 prosthesis templates is inserted through the window for determination of correct size and position. The corresponding implant is then inserted and secured with a hydroxylapatite shim. Thirty-five patients have been implanted for vocal fold paralysis, and 4 patients were implanted for soft tissue deficits or bowing. Thirty-one of 35 patients have reported subjective improvement (89%). Improvement was demonstrated in 13 of 15 (87%) patients with complete preoperative and postoperative objective voice function measurements. Complications include 1 implant extrusion and 1 case of airway obstruction secondary to edema. Preliminary results indicate that prefabricated hydroxylapatite implants are effective for medialization thyroplasty. Advantages include a readily available implant selection, rapid determination of correct size and position, and improved implant stabilization with a hydroxylapatite shim. PMID- 8239345 TI - Correlation between stroboscopy and electromyography in laryngeal paralysis. AB - Twenty patients with vocal fold motion impairment were reviewed to correlate the findings of electromyography (EMG) and stroboscopy. The causes of motion impairment were idiopathic, previous surgery with recurrent laryngeal nerve injury, neck and skull base trauma, and neoplasm. The EMG studies were analyzed to assess the status of innervation of the immobile vocal fold. The presence or absence of the mucosal wave prior to therapeutic intervention was determined with stroboscopic examination. Eight of 10 patients with EMG evidence of reinnervation or partial denervation were found to have mucosal waves, and 3 of 10 patients with EMG evidence of denervation were found to have mucosal waves. Six patients developed mucosal waves after surgical medialization, despite evidence of denervation by EMG criteria. These findings support the premise that tension and subglottic pressure, rather than status of innervation, determine the presence of the mucosal wave. PMID- 8239346 TI - Temporary nasosinusal drainage and lavage in chronic maxillary sinusitis. Statistical study on 847 maxillary sinuses. AB - We report on 498 patients suffering from chronic maxillary sinusitis verified by sinusoscopy through the inferior meatus and by computed tomography of the sinuses, which permitted also the diagnosis of coexisting chronic ethmoidal sinusitis. The patients were treated by temporary drainage through a polyethylene drain tube left in place at the end of the antroscopy for daily lavage with saline solution and daily instillation in the sinus of thiamphenicol glycinate hydrochloride (similar to chloramphenicol) and N-acetyl-L-cysteine. We followed up 847 maxillary sinuses during the lavages according to 2 parameters: 1) recovery of a clean washout product at the macroscopic examination and 2) recovery of normal ostial patency as measured by sinusomanometry. We concluded that 1) the association with ethmoidal sinusitis does not significantly modify the treatment duration, and 2) fewer than 4% of the maxillary sinuses are likely to recover normal parameters after the 23rd day of lavage, and it is therefore useless to continue this treatment much longer. Of the maxillary sinuses, 36.25% were not submitted to any surgery. The results gained by the lavage technique have to be protected from recurrence by the treatment of general, focal, or local causes. PMID- 8239347 TI - Esophageal stricture: an uncommon complication of foreign bodies. AB - Ingestion of foreign bodies is common in infants. As a rule, the infant is being observed and the event is witnessed. When it is unwitnessed, acute symptoms bring the problem to the attention of a physician. When the ingestion is not witnessed or symptomatic, its presence can only be surmised. The object can create injuries and chronic symptoms that are not typical for this accident. Here is reported an ingestion of a coin that went undiagnosed and untreated for more than a year. The result was a transmural esophageal injury with stricture formation. The diagnostic methods and transcervical repair are described. PMID- 8239348 TI - Lightning injury of the tympanic membrane. AB - A number of cases of otologic injuries by lighting strikes have been described in the otolaryngological literature. The mechanism of these injuries remains uncertain. We report 3 cases of lightning injury that presented to us. Analysis of these cases suggests that the mechanism of injury is direct conduction of electricity from the scalp to the soft tissues of the external auditory canal to the tympanic membrane. The conduits of the electrical surge are the subcutaneous blood vessels, smaller vessels being damaged more than larger vessels. Since the tympanic membrane central vessels are smaller than the canal vessels, the central area of the tympanic membrane would be most vulnerable, and this is seen clinically. A review of the literature supports this proposed mechanism of injury. PMID- 8239349 TI - Malignant external otitis in nondiabetic patients. AB - The purpose of this study is to point out that contrary to traditional belief, there is a small but significant group of nondiabetic patients with malignant external otitis. Thirty patients with a diagnosis of malignant external otitis were treated and followed up at the Department of Otolaryngology-Head and Neck Surgery, Beilinson Medical Center, between 1987 and 1991. Nine of these patients did not have clinical or laboratory evidence of diabetes. This study analyzes this group and concludes that the diagnosis of malignant external otitis should be considered by the treating physician in nondiabetic patients presenting with a Pseudomonas aeruginosa infection of the external ear canal. Severe pain and edematous closure of the canal, together with typical granulation tissue and failure to respond to medical treatment, are specific characteristics of this group. PMID- 8239350 TI - Sudden sensorineural hearing loss associated with slow blood flow of the vertebrobasilar system. AB - To determine the characteristics of sudden deafness associated with slow blood flow (SBF) within the vertebrobasilar arteries, we evaluated 57 patients with sudden deafness using magnetic resonance imaging (MRI). We detected SBF in 12 (21%) patients, predominantly men over 50 years of age. A second MRI performed in 5 patients 2 months after the onset of symptoms showed recovery of blood flow. All 12 patients complained of vertigo. Audiological and neurotologic tests suggested that hearing loss mainly involved the inner ear. Our findings suggest that unless central lesions are detected, headache, hypoesthesia of the external ear canal, and electronystagmographic abnormalities are signs of SBF. Because sudden deafness may recur in patients who have SBF, they should be monitored and treated to prevent recurrence. PMID- 8239351 TI - Improvements in respiratory and psychosocial functioning following total laryngectomy by the use of a heat and moisture exchanger. AB - A prospective clinical study in 61 patients was undertaken to investigate the subjective and objective influence of a heat and moisture exchanger (HME) on the respiratory and psychosocial problems following total laryngectomy. Although statistical comparisons failed to detect significant differences between the experimental and the control groups, there was a clear trend toward improvements in respiratory and psychosocial functioning in the experimental group. Analyses of differences over time within the HME user group showed significant reductions in the incidence of coughing, the mean daily frequency of sputum production, forced expectoration, and stoma cleaning. Significant improvements were also found in shortness of breath, fatigue and malaise, sleep problems, levels of anxiety and depression, and perceived voice quality. Pulmonary function tests showed significant improvements in inspiratory flow and volume values following use of the HME. This objective improvement in inspiratory pulmonary function reflects the decrease in sputum production reported by the patients. PMID- 8239352 TI - Localized amyloidosis of the larynx: evidence for light chain composition. AB - We report the biochemical characterization of amyloid fibrils from a patient with localized amyloidosis of the epiglottis and larynx. Biopsy specimens showed amorphous material consistent with amyloid deposits with a plasmacytic infiltrate. Both plasma cells and amyloid deposits stained positively by immunohistochemistry for kappa light chains. Amyloid fibrils were isolated. The major constituent resolved as a 13 kd band was sequenced and found to be consistent with a kappa 1 light chain. A tryptic digest was carried out and 3 tryptic peptides were sequenced defining the first 45 residues of the protein and residues 110 through 119. Four amino acid substitutions were found, 3 of which have not been described previously. This study defines the immunoglobulin origin of amyloid deposits in localized amyloidosis. The benign nature of localized amyloidosis suggests that a localized clone of plasma cells producing an amyloidogenic light chain may represent the pathogenetic mechanism of this disease, which appears to be a form of plasma cell dyscrasia. PMID- 8239353 TI - Inverted papilloma of the middle ear. AB - Inverted papilloma is a benign neoplasm confined almost exclusively to the sinonasal tract. We present the first known case report of an inverted papilloma arising from the middle ear. In this paper we discuss the pathophysiology and review the literature of this interesting clinical entity. PMID- 8239354 TI - Technetium 99m dimercaptosuccinic acid scintigraphy in medullary carcinoma of the thyroid. PMID- 8239355 TI - Pure second arch syndrome. PMID- 8239356 TI - Wegener's granulomatosis and antineutrophil cytoplasmic autoantibodies. AB - Serologic tests for antineutrophil cytoplasmic autoantibodies can serve as markers for a number of necrotizing vasculitides and other inflammatory disorders. In the case of Wegener's granulomatosis, an immunofluorescent cytoplasmic pattern and immunoassay for proteinase 3 are quite specific for the disease and are capable of serving as additional criteria for diagnosis. As with any laboratory test, however, results are to be interpreted in the whole clinical and pathologic framework of the disease. PMID- 8239357 TI - [Duplicate publication]. PMID- 8239358 TI - [Basal cell carcinoma of the nose]. AB - The purpose of this study of 81 patients with basal cell carcinoma (BCC) of the nose was to present the oncological and cosmetic results of surgical treatment and compare these results with those of other possible treatments. MATERIAL AND METHOD: We report a series of 81 cases of histologically proven BCC of the nose located chiefly on the alae nasi and on the lower end of this organ; 42 p. 100 of the tumors had previously been treated and had recurred. The patients' mean age was 63 years, and the shortest follow-up was 3 years. Excision of the tumor under simple or reinforced local anaesthesia was complete in 88 p. 100 of the cases, incomplete or borderline in 12 p. 100 and systematically repeated. Extemporaneous histological examination was performed in 18 p. 100 of the cases. The operative lesion was repaired with a graft or a flap. There was no postsurgical treatment. RESULTS: The recurrence rate was 4 p. 100 with a minimum follow-up of 3 years. The cosmetic result was good in 78 p. 100 of the patients. DISCUSSION: Numerous treatments have been used against BCC of the nose, the results, advantages and disadvantages of each of these treatments are given below: 1. Cryosurgery. The problem with this method is that it is relatively difficult to perform and requires reliable operators. The cure rate is similar to that of other treatments. 2. Chemotherapy is not frequently used. 3. Electrocoagulation. Contrary to the conventional excision, this method precludes all histological controls, and the common idea of good oncological results is now being revised. 4. Radiotherapy. The recurrence rate varies from 7 to 11.8 p. 100 with fair cosmetic results. It requires numerous sessions, cannot be repeated in case of recurrence and complicates the surgical treatment. In addition, there is a long term risk of radiodystrophy. 5. Curietherapy by local implantation of 192Iridium has a recurrence rate of 2.5 to 7 p. 100. This treatment requires hospitalization and is costly. It is indicated in cases of complex surgery, difficult conventional radiotherapy and above all, sclerodermatous BCC. 6. Surgery has a recurrence rate which varies from 3.7 to 12.6 p. 100. Recurrences are due to insufficient excision, and this underlines the value of histological control which can be done only at surgery. It is a rapid and ambulatory treatment, usually performed under local anaesthesia. The cosmetic result is good (85 to 90 p. 100) and the patients must be made aware of this preoperatively with some degree of reliability, depending on age, location, size of BCC and on the surgical technique utilized. CONCLUSION: Surgical treatment of nasal BCC is safe, effective and rapid, with good oncological and cosmetic results, and it has many advantages over the other treatments. PMID- 8239359 TI - [Melanoma and primary hyperparathyroidism]. AB - The authors present 3 cases of melanoma associated with a parathyroid adenoma discovered by routine measurement of blood calcium levels. The melanomas were differently located and had different histological degrees. Hypercalcaemia is an infrequent complication of melanoma. According to data found in the literature, it is most often consecutive to bone metastases. The other causes of hypercalcaemia are metastatic extension of melanoma to the parathyroid glands, secretion of parathormone-related peptide (PTH-RP) and the actions of prostaglandins, vitamin D and the osteoclast-activating factor (OAF). Primary hyperparathyroidism due to adenoma has seldom been described associated with melanoma. PMID- 8239360 TI - [Acral erythema after allogeneic graft of the bone marrow]. AB - Acral erythema (AE) is a painful, erythematous bullous eruption of the palms and soles which is chemotherapy-induced. To the numerous chemotherapies which induce AE we must add, perhaps, a new drug, Vepeside. AE is followed by graft-versus host-disease in all patients receiving bone marrow transplantation. AE and early GVH disease being very similar, we discuss the differential diagnosis which can be very difficult. PMID- 8239361 TI - [Acute generalized exanthematous pustulosis in a child and epicutaneous tests with mercurials]. AB - Diffuse exanthema with fever followed by superficial nonfollicular aseptic pustules was observed in an 11-year old boy after conducting a thiomersal epicutaneous test and manipulation of a broken medical thermometer. The patient healed within two days without any treatment. The initial responsibility of mercury and its derivatives, and the primary sensitization and relevance of the epicutaneous tests are discussed. Mercury and its derivatives should be limited as much as possible in medicine and cosmetology. PMID- 8239362 TI - [Cutaneous and nail infections caused by Hendersonula toruloidea]. AB - A case of cutaneous and nail infection due to Hendersonula toruloidea (HT), in a Senegalese woman who had been living in France for 7 years is reported. This patient presented with keratotic lesions of the palms and distal and lateral onycholysis. Culture of clinical specimens on Sabouraud's medium without cycloheximide resulted in the isolation of HT. This rare fungal infection is often misdiagnosed. PMID- 8239363 TI - [Psoriatic onycho-pachydermo-periostitis]. AB - Psoriatic-onycho-pachydermo-periostitis is a particular form of psoriatic arthropathy recently described, which combine psoriatic onychosis, thickening of the distal soft tissues and osteo-periostitis of the distal phalanx without lesion of the interphalangeal joint. Biological examinations are normal. Radiological lesions show a phalanx condensation which gives a spicule aspect. Two cases of onycho-pachydermo-periostitis are described. We report the first one with all the fingers and toes concerned. PMID- 8239364 TI - [Systemic urticaria disclosing post-vaccine lupus tuberculosis]. AB - The authors report a case of association between lupus vulgaris and urticarial vasculitis after BCG vaccination. Lupus vulgaris is a rare classical complication of BCG vaccination. Anaphylactoid reactions after BCG are very rare. This is the first report of association between the two diseases after BCG and the third report of urticaria after BCG. The pathogenesis of this is discussed. PMID- 8239365 TI - [Squamous erythema annulare centrifugum]. AB - A case of squamous erythema annulare centrifugum has been observed. It was characterized by striking clinical features and peculiar histological findings, including necrosis of keratinocytes. A search for causative factors was negative. Treatment with tar ointment (Brocq) rapidly resulted in dramatic healing of the lesions; however, the usual features of annular psoriasis were absent in our case. PMID- 8239366 TI - [Blue rubber bleb nevus]. AB - Blue Rubber Bleb Naevus (BRBN) is characterized by the presence of bluish haemangiomas on the skin and in the digestive tract. We report a new familial case of this rare entity. Analysis of this female patient's pedigree confirmed that BRBN is transmitted as an autosomal dominant trait. In this particular case gastrointestinal haemangiomas appeared over a 4-year period. Although all treatments of this disease have proved disappointing on the skin as well as in the digestive tract, we insist on the need for a regular and prolonged surveillance. PMID- 8239367 TI - [Telematic information on sexually transmitted diseases and AIDS]. PMID- 8239368 TI - [Enzymatic debridement of leg ulcers using papain]. PMID- 8239369 TI - [Is paraneoplastic pemphigus a type of pemphigus?]. PMID- 8239370 TI - [Case for diagnosis: lichen striatus]. PMID- 8239371 TI - [Marine skin diseases of the Brittany coast]. PMID- 8239372 TI - [Signal transduction in dermoepidermal cells: application to psoriasis]. PMID- 8239373 TI - [How to treat a psoriatic nail?]. PMID- 8239374 TI - [Structural abstracts. Informative abstracts]. PMID- 8239375 TI - [Anogenital papillomavirus lesions in humans with or without HIV infection. Comparison of colposcopic, histopathological and virological results]. AB - Anal and genital lesions caused by human papilloma virus (HPV) may be associated with severe dysplasia and cancer, chiefly in cases of "high risk" HPV types 16, 18, 31, 35, 51. The frequency of HPV infections and the severity of genital cancers seem to be increased in patients with human immunodeficiency virus (HIV) infection. PATIENTS AND METHOD. The distribution of different HPV types was compared with the anatomical and clinical features of the lesions in two populations, one HIV+ (n = 40) and the other HIV- (n = 48), who had anal and genital lesions. The HPV DNA was determined by molecular hybridization in situ, using biotinylated probes which recognized HPV types 6/11, 16/8 and 31/35/51 on 99 lesions. RESULTS. HIV+ subjects differed from HIV- subjects in that a higher proportion of them had anal lesions (50 p. 100 vs 10 p. 100) and condyloma latum (80 p. 100 vs 50 p. 100). Koilocytosis without dysplasia was more often found in HIV- subjects (12.5 p. 100 vs 55 p. 100). Conversely, dysplasia was more frequent among the lesions of HIV+ subjects: grade I 39.5 p. 100 vs 17.5 p. 100; grade II 25 p. 100 vs 4 p. 100; grade III 12.5 p. 100 vs 0 p. 100. Koilocytosis was preferentially associated with condyloma acuminatum. In HIV+ subjects the DNA of HPV, detected in 73 p. 100 of the lesions, was "high risk" HPV DNA in 86 p. 100 of the cases, whereas in HIV- subjects 51 p. 100 of the samples were positive in hybridization, and 61.5 p. 100 had "low risk" HPV DNA. In subjects of all groups "high risk" HPV was found in dysplastic lesions. CONCLUSION. HIV seropositive subjects show an imbalanced distribution of HPV with predominance of "high risk" HPV. This suggests that immunodepression encourages infection by this oncogenic virus, thereby contributing to the frequency of cancer in HIV+ subjects. PMID- 8239376 TI - [Pityriasis rotunda]. AB - Pityriasis rotunda (PR) is a well-known skin disease in Japan, but it is seldom observed in Europe. Because it is often associated with an underlying, sometimes malignant disease, it is regarded by some authors as a clinical form of acquired ichthyosis. We report the case of a 20-year old male Congolese presenting with PR and ichthyosis vulgaris. This rarely described association raises the problem, discussed here, of whether PR belongs to ichthyosis vulgaris. PMID- 8239377 TI - [Mycobacterium kansasii cutaneous infection]. AB - The authors report a case of cutaneous infection caused by Mycobacterium kansasii in an immunocompetent woman. The mycobacterium was identified after a search for mycolic acids and the species-specific phenol-glycolipid K1. As minocycline followed by ciprofloxacin were ineffective, a conventional antituberculous treatment was prescribed and was fully successful. PMID- 8239378 TI - [Colonic adenocarcinoma and scleroderma]. AB - The association of cancer and connective tissue disease is well known, the most frequent being certainly with dermatomyositis. The association cancer and PSS is more controversial. The incidence of neoplasia in that group seems to be comparable with the general population but the proportion of certain types of cancer is different, and the temporal relationship with the apparition of symptoms of PSS is stunning. The hypothesis actually in favor is an imbalance of the immune system, which cause the diminution of the immune surveillance and the apparition of cancer and a concomitant dysregulation of the system, causing the fibrosis of the PSS. We describe a 75 year-old white female who developed a colic adenocarcinoma; she also had, concomitantly, a systemic scleroderma, with sclerodactyly and pulmonary fibrosis. The patient corresponded to the criteria of the American Rheumatism Association for progressive systemic scleroderma (PSS). The prognosis of patients with PSS depends on their systemic involvement but also, we believe, in the more aged group, on the apparition of a neoplasia. PMID- 8239379 TI - [Microcapillary hemangioma]. AB - A cutaneous microcapillary hemangioma was discovered in a young woman 4 months after a change of oral contraceptives. This recently individualized benign tumor is made of well differentiated capillaries without cellular atypias, located in the reticular dermis. It arises in a few weeks in young subjects, sometimes after contraceptive change or during pregnancy. Sexual hormonal imbalance may explain some cases. The histological appearance of microcapillary hemangioma is remindful of microvenular hemangioma, more recently described, which also arises in a short time in young patients. PMID- 8239380 TI - [Erythroderma disclosing transformation of refractory anemia with excess of blasts into lymphoblastic leukemia]. AB - We report a case of erythroderma revealing lymphoblastic leukaemia occurring immediately after myelodysplasia. The patient was an 87-year old man admitted for poor general condition, lymph node enlargement and pruriginous oedematous erythroderma. Laboratory examinations showed tricytopenia, bone marrow invasion by lymphoblasts in a myelodysplastic environment of the refractory anaemia type with excess of blasts, and a cutaneous lymphocytic infiltrate with non-blastic lymph node reaction. Owing to its morphological and immunohistological features, the lymphocytic infiltrate was regarded as non-specific. We could not help comparing this case to the other non-specific cutaneous lesions of refractory anaemia which usually reflect transformation into leukaemia. PMID- 8239381 TI - [Pityriasis rotunda. Review of the literature]. PMID- 8239382 TI - [Cyclosporine and Darier's disease]. PMID- 8239383 TI - [Friedrich Daniel von Recklinghausen (Gutersloh 2 December 1833--Strasbourg 25 August 1910)]. PMID- 8239384 TI - [Apropos of the article by Cesarini JP, Muel B: Dermatologic risks of quartz halogen lamps]. PMID- 8239385 TI - [A case for diagnosis: ungueal psoriasis]. PMID- 8239386 TI - [Skin diseases and enthesiopathies]. PMID- 8239387 TI - [Atopic dermatitis and delayed respiratory hypersensitivity]. PMID- 8239388 TI - [Immunosuppression, virus, herpesviridae and cutaneous cancers]. PMID- 8239389 TI - [Statistic study of 5,473 somatotropin secretion stimulation pharmacologic tests (n=9). Proposed weighting coefficient]. AB - A total of 5,473 pharmacological provocative growth hormone release tests were carried out in 3,143 children. Mean age was 9 years 9 months (range 3-16 years) and mean bone age was 7 years 6 months (range 2-14 years). Tests were of 9 different types: 1) arginine (n = 625); 2) clonidine (n = 339); 3) insulin (n = 198); 4) ornithine (n = 162); 5) insulin + arginine (n = 203); 6) clonidine + betaxolol (n = 2,003); 7) L-dopa (n = 685); 8) glucagon = propranolol (n = 443); 9) glucagon + betaxolol (n = 815). All growth hormone determinations were performed using the same radioimmunoassay. Distribution of values obtained with each test was gausso-logarithmic. Mean peak levels with their 95% confidence limit were as follows: 1) 10.2 and 0.45; 2) 11.5 and 0.7; 3) 11.8 and 0.8; 4) 14.2 and 1.2; 5) 14.3 and 0.9; 6) 15.7 and 1.1; 7) 19.8 and 2.1; 8) 20.8 and 2.3; 9) 21.0 and 2.5. These data indicate low specificity, with up to two-fold differences in mean peak levels from one test to another; proportions of peaks under 10 ng/ml ranged from 29% to 69%. Thus, the rate of patients diagnosed with growth hormone deficiency may vary substantially according to the test used. To reduce these discrepancies, we suggest adjustment of test results using a weighting coefficient of 1) 1.9; 2) 1.48; 3) 1.4; 4) 1.16; 5) 1.06; 6) 1.01; 7) 0.73; 8) 0.69; 9) 0.66. PMID- 8239390 TI - [Discovery, in neonatal screening, of 2 families carrying a syndrome of generalized resistance to thyroid hormones]. AB - The syndrome of generalized resistance to thyroid hormones is being increasingly diagnosed, albeit often belatedly. In the two families described herein the diagnosis was established when moderately elevated thyrotropin levels were found upon neonatal screening of a family member. The family studies identified other affected members with a pattern indicating autosomal dominant inheritance. Clinical and laboratory findings in the neonates were consistent with normal thyroid function and no treatment was given. In one neonate, fibroblast nuclear receptor studies failed to detect decreased affinity for triiodothyronine, a finding reported in most previously published cases; the mutation in this patient was different from the one described in 1989 by Sakurai et al., consistent with the known genetic heterogeneity of this syndrome. It has been suggested recently that treatment of affected neonates with large doses of thyroid hormones is safe and effective in ensuring normal growth. The neurodevelopmental effects of this treatment are unknown. Early treatment is possible when the syndrome is detected neonatally. We therefore advocate routine T4 assays in neonates with moderately elevated TSH levels. PMID- 8239391 TI - [Bone demineralization and elevation of serum osteocalcin concentrations in young children with hyperthyroidism]. AB - Bone mineralization and serum osteocalcin level were evaluated in 15 children with Grave's disease. Two groups were constituted according to the presence (group I: n = 9) or absence (group II: n = 6) of a severe bone demineralization. A spontaneous fracture and a collapsed vertebra were found in one group I patient. Patients in group I were younger than in group II (8.3 +/- 4.9 vs 11.5 +/- 4.3 yrs). One patient in group II and six in group I were prepubertal with advanced bone age and increased growth velocity. Osteocalcin measurement (Oc) was performed in 10 patients (group I: n = 6; group II: n = 4) at the time of biological hyperthyroidism. The six patients with bone demineralization had elevated Oc levels. In group II, two patients had normal Oc levels and two had elevated Oc levels. In treated patients with good control of hyperthyroidism, all group II patients except one, had normal serum Oc levels and bone mineralization remain normal (n = 5) after 0.6 to 4.6 yrs of follow-up. In group I patients, although height velocity was normal, elevated (n = 4) or slightly elevated (n = 1) serum Oc levels and severe bone demineralization (n = 7 cases) persisted after 0.5 to 3 yrs of good control of the hyperthyroidism. Although the method used for measuring bone mineralization is potentially less precise than bone densitometry and not all the patients had serum osteocalcin measurements at the same time of the illness, our results emphasize that skeletal demineralization may be particularly marked in young children with Grave's disease and should be carefully evaluated. PMID- 8239392 TI - [Late diagnoses of 21-hydroxylase deficiencies in children (after the age of 3 years]. AB - To evaluate the heterogeneity of 21-hydroxylase deficiency with delayed symptoms, clinical and laboratory findings at presentation in 29 patients whose first symptoms occurred after three years of age were analyzed retrospectively. In 12 patients, these data were confronted with the results of molecular CYP21B gene analysis. Age at onset was 7 years on average and was comparable in boys and girls. Premature puberty was the most common presenting symptom [n = 24], whereas hirsutism, clitoral enlargement, and menstruation disorders were less frequent. Six cases were diagnosed as the result of routine studies of family members of index patients. The bone age over statural age ratio was greater than 1 in 19 of the 27 patients. Baseline 17-OH-progesterone levels were elevated in 22 of the 27 patients; magnitude of the elevation varied widely. Levels of 17-OH-progesterone after stimulation with immediate-action tetracosactide were closely correlated with baseline values and established the diagnosis in doubtful cases. Four patients had post-stimulation 17-OH-progesterone levels under 10 ng/ml, suggesting that were heterozygous for the disease. An important finding was that the magnitude of the devation in 17-OH-progesterone was not clearly correlated with clinical findings at presentation (age at onset, growth rate, advance in bone age). Molecular CYP21B gene analysis performed in 12 patients disclosed a homozygous 281 Val Leu mutation in 6 cases. This is the most commonly reported mutation in delayed onset forms. Two patients were heterozygous for the 281 Val Leu mutation and had an allele associated with severe disease, suggesting that the least severely affected chromosome governed clinical presentation of the disease. One boy had an allele associated with neonatal onset on both chromosomes; molecular analysis indicated a risk of antenatal masculinization of female fetuses in this family. This study showed that delayed onset 21 hydroxylase deficiency is a heterogeneous entity and that molecular analysis is essential to genetic counseling. PMID- 8239393 TI - [Growth hormone administered to non-growth hormone deficient, small girls: echographic aspect of the gonads and uterus]. AB - The effect of growth hormone (GH) treatment on prepubertal gonads is controversial especially with regard to the risk of precocious puberty. Ultrasound assessment of ovarian volume, follicle size, and uterine growth was performed in 20 premenarcheal girls (8.0 +/- 2.6 years) receiving growth hormone (GH) for short stature (-2.8 +/- 0.4 SD) not related to growth hormone deficiency or Turner syndrome. Mean GH dosage was 1.0 +/- 0.4 IU/kg/week and mean duration of treatment at evaluation was 16.3 +/- 8.9 months. All patients underwent real time ultrasonography of the pelvic organs and ten subjects also had color Doppler studies of the ovarian and uterine arteries. Ultrasound findings were similar to those reported in normal prepubertal girls. Mean uterine length (29.1 +/- 7.5 mm) and volume (1.23 +/- 0.86 ml) were correlated with age but not with dosage or duration of GH treatment. Ovarian volumes was within the normal age-specific range in all patients except a 7.9 year old girl with substantially enlarged ovaries (4.7 ml) but no evidence or precocious puberty. Ovarian follicles were found in five girls; they measured less than 9 mm in diameter in every case except one (13 mm follicle in an 11-year-old). Blood flow in the ovarian arteries was seen on 5 of the 10 color Doppler studies and was not correlated with dosage or duration of GH treatment. Administration of GH to non-GH-deficient girls did not substantially affect the internal genital organs. It remains uncertain whether the single case of ovarian enlargement seen was related to GH treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239394 TI - [Klinefelter syndrome in 1993. Results of a multicenter study on 58 cases and review of the literature]. AB - A retrospective multicenter study found 58 cases of Klinefelter syndrome of which 23 (39%) were diagnosed before puberty. Although as common as Down syndrome, Klinefelter syndrome is underdiagnosed and often recognized only in adulthood. Suggestive manifestations in infants, children, and teenagers include facial dysmorphism, micropenis, and delayed speech and should lead to examination of the karyotype. Early recognition of Klinefelter syndrome could be achieved by routinely measuring the size of the testes in school-boys aged 11 to 15 years and performing a karyotype in boys with a volume of less than 2 ml. Early psychological and educational support and testosterone replacement therapy initiated at onset of puberty may lead to improved social and academic outcomes. PMID- 8239395 TI - [Sertoli-Leydig tumors in children. 2 case reports]. AB - Sertoli-Leydig tumors stem from the mesenchyma and sexual cords of the embryonic gonad. Two cases are reported. One manifested as symptoms of virilization in a 12 year old girl. The other patient developed adnexal torsion at the age of five years. Pelvic ultrasonography visualized the tumor in both cases. Increased production of ovarian androgens suggested the diagnosis in the first case. Histological studies disclosed intermediate differentiation in the first case and tubular differentiation in the second. These tumors usually exhibit low-grade malignancy and unilateral salpingo-oophorectomy ensures recovery in most instances. PMID- 8239396 TI - [Screening in the school milieu, at 4 years old, for hypercholesterolemia]. AB - Four-year-old schoolchildren with a positive family history for atherogenic dyslipidemia and/or clinical atheroma before 55 years of age were screened for hypercholesterolemia. Investigations included determination of serum levels of total cholesterol, triglycerides, HDL cholesterol, apolipoprotein A1, apolipoprotein B, and Lp(a); an agarose lipidogram; acrylamide gradient electrophoresis; and determination of LDL composition by ultracentrifugation. Normal values were defined as values under the 90th centile, i.e., 1.97 g/l for total cholesterol, 0.89 g/l for triglycerides, 1.36 g/l for LDL-cholesterol, and 1.26 g/l for apolipoprotein B. Among 3,565 children routinely evaluated at 4 years of age, 525 (16.2%) had a positive family history; of these, 72 underwent lipid investigations. Eight children (11%) had hypercholesterolemia type IIA, eight had a variety of lipid disorders, and 14 (20.6%) had increased Lp(a) levels as an isolated anomaly or concomitantly with an atherogenic dyslipidemia. Because Lp(a) is a cardiovascular risk factor independent from total cholesterol levels, we believe this parameter should be determined in high risk children. PMID- 8239397 TI - [Cushing syndrome in children]. AB - In pediatric patients, endogenous Cushing syndrome is an infrequent condition almost always due to one of two conditions. 1) Adrenal gland tumors account for 70% of Cushing syndromes in young pediatric patients. They cause rapidly progressive hypercorticism not due to increased ACTH production (elevated plasma and urine cortisol levels, very low ACTH and LPH levels unchanged by dexamethasone, metyrapone or CRH). Imaging techniques determine the side and spread of the tumor and look for metastases. Following surgical removal, patients with indicators of malignant disease (tumor weight above 30 g, extracapsular spread or metastases, independently from pathological data) are given op'DDD. 2) Cushing disease occurs in peripubertal patients and causes overweight with delayed statural gain. ACTH production is increased (positive dexamethasone suppression test and provocative metopirone and CRH tests) as a result of a pituitary adenoma which should be looked for by magnetic resonance imaging and whose removal ensures recovery in 50% of cases. Other therapeutic tools include op'DDD, radiation to the pituitary, and bilateral adrenalectomy as the last resort given the high risk of post-adrenalectomy pituitary tumor (50% of pediatric patients). Other causes are exceedingly rare: primary nodular hyperplasia of the adrenal glands and production of ACTH by a nonpituitary tumor. Corticosteroid treatment is the most common cause of Cushing syndrome in children. PMID- 8239398 TI - [Endocrinal sequela in bone marrow transplantations during childhood and adolescence]. AB - Bone marrow transplants are being increasingly used to treat a broadening spectrum of serious pediatric conditions including hematologic, metabolic, and immune disorders. A common adverse effect is slowing of statural growth which, according to the author's experience, reaches 0.4 SD a year on average in the highest risk group with progressive graft-versus-host disease after whole body irradiation and transplantation of an allograft. Growth was normal in autograft recipients treated by moderate-intensity chemotherapy. High-dose chemotherapy, in particular with busulfan and cyclophosphamide, has been reported to cause growth retardation. The mechanism of failure to gain height is complex. Partial growth hormone deficiency is rare. Relative resistance of growth plates is the most likely mechanism. The role and efficacy of growth hormone replacement therapy is unclear. Conditioning regimens can affect ovarian and testicular function. Peripheral thyroid resistance can occur after whole body irradiation and transplantation of an allograft. The high rate of growth and endocrine disorders warrants close monitoring including full evaluations of growth and pubertal development as well as periodic endocrinologic investigations. PMID- 8239399 TI - [Role of magnetic resonance imaging in non-neoplastic hypothalamo-hypophyseal pathology]. AB - This study was designed to evaluate the diagnostic, therapeutic, and pathogenetic data provided by magnetic resonance imaging (MRI) in nonneoplastic hypothalamo pituitary disorders. After determination of age-specific pituitary heights, 46 children with idiopathic growth hormone deficiency (GH peak < 8 ng/ml) were studied. Twenty-nine patients (group I) had pituitary stalk interruption syndrome and 17 (group II) had no anatomic abnormalities. Age-specific pituitary height was decreased by more than 2 SDs in all group I patients versus only 60% (10/17) of group II patients. The GH deficiency was transient in 4 of the 7 children with normal pituitary findings. Forty-seven girls with breast development before eight years of age were also studied: age-specific pituitary height was normal in all girls with premature thelarche and 68% of girls with mild form of central precocious puberty (CPP). Conversely, in 70% of girls with evolutive CPP, age specific pituitary height was increased by more than 2 SDs. These data show that MRI is useful for the diagnosis of pituitary insufficiency and that multiple anterior pituitary deficiencies can be expected in patients with anatomic abnormalities. MRI is of diagnostic and prognostic usefulness in CPP and, therefore, is of assistance in making therapeutic decisions. PMID- 8239400 TI - [Hypothalamic syndromes. Review of clinical and endocrinal semiology]. AB - A case of hypothalamic dysfunction in a girl with a twelve-year follow-up is reported. Onset occurred at the age of three with severe obesity, hypothermia, hypersomnia, and lethargy. Somatotropic, gonadotropic, and thyrotropic hormones were low, whereas prolactin was increased. Imaging techniques failed to disclose any lesion of the hypothalamus or pituitary. Clomipramine improved the vegetative disorders. The literature on clinical and hormonal disorders of hypothalamic dysfunction is reviewed. PMID- 8239401 TI - [Efficacy and tolerance of a local application of phenazone and chlorhydrate lidocaine (Otipax) in infants and children with congestive otitis]. AB - The efficacy and safety of ear drops containing phenazone and lidocaine hydrochloride (Otipax) for the treatment of congestive myringitis were evaluated in 18 infants and children aged 1 to 10 years. Relief of pain was evident 5 minutes after instillation and significant after 15 to 30 minutes. Serial photographs of the tympanic membrane demonstrated prompt improvement of inflammation. Congestion was significantly reduced after five minutes and overall ear drum color was significantly improved after 15 to 30 minutes. No adverse effects were recorded. These data suggest that Otipax is effective and safe for the treatment of painful congestive myringitis in infants and children. PMID- 8239402 TI - [Post-traumatic stress syndrome in children]. AB - Posttraumatic stress disorder is a form of anxiety disorder which is poorly known in children and manifests as a series of symptoms occurring after an extraordinary event outside the range of usual human experiences and responsible for a feeling of terror. Virtually pathogmonic symptoms include re-experiencing the event through play and ceaselessly repeated behaviors (re-enactments), cognitive distortions when relating the facts (chronological errors, belief in omens), changes in attitudes towards others and life in general, and neurovegetative hyperactivity (hypervigilance, startle responses, difficulties controlling impulses). Although adequate follow-up data are lacking, the disorder can probably become chronic in children, as in adults. This nosographic entity raises the theoretical issue of the roles of life events and individual vulnerability. A number of factors may either "predispose" or "protect" the child: degree and duration of exposure to the trauma, nature of the event, preexistence of psychiatric disorders, level of cognitive development, sex, degree of social support, and containing or noncontaining attitude of the family. Treatments advocated in adults (pharmacotherapy, cognitive and behavioral therapies, group therapy) remain to be used and evaluated in children. PMID- 8239403 TI - [Depressed mothers: the impact of depression on early interactions. An analysis of Anglo-Saxon studies]. AB - Maternal depression remains a public health problem as indicated by many studies focusing on depression in mothers of young children. Although the high prevalence of depression in mothers of infants and young children is now a recognized fact, the detection and management of maternal depression in everyday practice still raises significant problems. This initial review centers on studies providing diagnostic guidelines. The problem of maternal depression and of its impact on the offspring is relevant to the issue of how qualities, abilities, and vulnerabilities are transmitted from one generation to the next. Psychoanalysts, infant psychiatrists, and developmental psychologists show great interest in this field. The current review was restricted to recent Anglo-Saxon studies on depression-related changes in early maternal behavior. The most striking findings are as follows: although depression affects maternal behavior overall, there is considerable variation across mothers; timing alterations (in terms of micro and macro sequences) in mother-child interactions occur in every case and are among the obstacles to harmony and synchronization; subtle alterations in the mother's response to her baby's signals preclude flexibility and anticipation. PMID- 8239404 TI - [Scales and questionnaires for the evaluation of behavior disorders in children]. AB - The need for quantitative clinical evaluation tools for use in child psychiatry is obvious. Behavior rating scales are useful for comparing clinical and laboratory data, monitoring the effects of treatments, and enhancing communication between clinicians and investigators. The methodological principles used to construct and validate such tools are described. This approach is of benefit in most psychiatric disorders of children and adolescents. The questionnaires and scales most widely used throughout the world and available in French are reviewed. The advantages and drawbacks of evaluation scales in everyday practice and in research are discussed, with the BSE-A scale (Behavior Summarized Evaluation of Austism) as an example. PMID- 8239405 TI - [Experimental principles of hyperactive behavior in children]. AB - Neurophysiological studies have demonstrated that processing of information is defective in hyperactive children. Converging lines of evidence suggest that both perception and attention are normal in these patients. In contrast, there are alterations in motor processes and in the regulation of energy resources used for motor output. The clinical and therapeutic implications of clinical research data are discussed. PMID- 8239406 TI - [Attempted suicide by defenestration in children and adolescents]. AB - In adults, suicidal attempts by jumping from elevated sites have been extensively studied and usually occur in patients with major psychiatric disorders such as schizophrenia or melancholia. Among the children and adolescents admitted to the Robert Debre Hospital in Paris for attempted suicide from 1989 through 1992, 8 (1 boy and 7 girls) had jumped out of a window. Ages ranged from 11 1/2 years to 15 years. Two patients had depression but in the six other cases there was no evidence of a psychiatric disorder. The suicidal attempt occurred after a prolonged period of conflicts within the family. Management of such patients proved difficult because of the presence of severe injuries and above all of massive denial on the part of the patient and family. PMID- 8239407 TI - [A prospective study on the behavior of pediatric intensive care unit nurses, concerning the care given to suicidal children]. AB - Pediatric intensive care units use sophisticated medical technology and are staffed by deeply committed nurses who are subjected to significant psychological stress. This stress varies with the type of patient and influences the style and quality of care. With this respect, children and adolescents admitted after a suicidal attempt are considered catalysts. However, there have been no systematic studies of how pediatric intensive care nurses respond emotionally to their interactions with these patients. This epidemiological study conducted in five Parisian Teaching Hospital pediatric intensive care units used a specially designed questionnaire to evaluate nurses' responses on the basis of style of care. Children under 16 years of age admitted after attempted suicide were studied comparatively with same age children admitted for status asthmaticus or encephalopathy with seizures. Results highlighted the differences in nurses' psychological responses to these situations and their difficulties in interacting with patients. This study provides strict methodological guidelines for investigating an issue often discussed emotionally or on the basis of anecdotal data. PMID- 8239408 TI - [Attention deficit disorder and anxiety disorders: a co-occurrence study]. AB - Recent epidemiological studies have evaluated the concomitant occurrence of several disorders in children. Over the last few years there have been reports from Anglo-Saxon investigators that many children with attention deficit disorder hyperactivity also have anxiety disorders. This is the first French study addressing this issue. The prevalence and types of anxiety disorders were determined in 50 children with attention deficit disorder-hyperactivity using a questionnaire and a semi-structured interview. Over 40% of subjects had at least one concomitant anxiety disorder meeting DSM-III-R criteria. Hyperanxiety and separation anxiety were the most common anxiety disorders. Among patients with anxiety, over 40% had more than one anxiety disorder. These data are consistent with earlier Anglo-Saxon reports. Symptoms of anxiety which discriminated hyperactive children with and without anxiety disorders were identified. PMID- 8239409 TI - The presentation of personalities in plastic surgery. AB - This research was of 133 plastic surgery patients over a 1.5-year period. Data compilation and assessment began with the initial consultation and continued through the postoperative period of care. Patients were categorized according to their similarity to the personality disorder descriptions found within the Diagnostic and Statistical Manual of Mental Disorders, third edition revised, of the American Psychiatric Association. In addition, each patient was evaluated on 13 separate psychological and social factors. It was found that many of the factors that precede a favorable plastic surgery experience, for both patients and physician, are the same characteristics disproportionately represented in various personality types. Objective assessments of surgical outcome were not found to be necessarily predictive of a patient's ultimate satisfaction. This research identified those personality types most likely to be dissatisfied, along with those personalities who tended to withhold their approval of an excellent surgical result. This research provided both statistical and qualitative analysis of the impact of personality, as well as several psychological and social variables, on plastic surgery. Numerous suggestions have been offered for successful patient care as derivative of personality type. The impact of patient and physician personality upon medical malpractice was explored. Reference summaries have been provided that describe and categorize patient behavior according to personality. PMID- 8239410 TI - The black American nose. AB - A study of 201 black American noses has led us to divide the black American nose into three groups, i.e., Groups A, B, and C, which we have called the "African," the "Afro-Caucasian," and the "Afro-Indian." Belonging to the African group (Group A) were 44% of the noses, 37% were Afro-Caucasian (Group B), and 19% were Afro-Indian (Group C). Fifty-three percent of the African noses had a concave dorsum, whereas only 10% of the Afro-Caucasian and 8% of the Afro-Indian group had a concave dorsum. On the other hand, 36% of the Afro-Caucasian group had a hump as did 63% of the Afro-Indian group. Only 18% of the African group had a hump. The most common nostril types were type IV (20%), type V (27%), and type VI (25%), the distribution of which varied with the nasal type. Anthropometric measurements showed variations according to type, with the African noses being the shortest and widest, the AFro-Caucasian the narrowest, and the Afro-Indian being the longest. Cadaver dissection showed that the alar cartilages varied from small and thin in the African group to large and thick in the Afro-Indian. All were covered by a heavy layer of fibrofatty tissue. The value of this grouping in the evaluation and surgery of the black American nose is discussed. PMID- 8239411 TI - Osseointegrated titanium implants for the attachment of facial prostheses. AB - The ability to penetrate skin permanently with posts of pure titanium has led to a dramatic breakthrough in the attachment of facial prostheses. The posts are attached to screws of titanium that are completely integrated in the underlying bone, without any intervening soft tissue. This precise and rigid fixation permits rehabilitation of patients with complex defects that would be otherwise untreatable. PMID- 8239412 TI - Biointegrated hydroxylapatite-coated implant fixation of facial prostheses. AB - This pilot study reports the 2-year follow-up on hydroxylapatite-coated nonscrew titanium implants (Integral) used for fixation of auricular and orbital prostheses. Fifty-four implants have been inserted in a two-stage procedure for auricular reconstruction in 12 patients and orbital reconstruction in 2 patients. Clinically solid integration has occurred with 51 implants in 13 patients, three implants being lost in previously irradiated bone where soft tissue coverage was never adequate. Early soft tissue reaction occurred in those younger patients and where multiple previous surgery had occurred in the same area. Conservative local treatment produced resolution of this in all patients. Stable retention of cosmetically acceptable orbital prostheses was achieved with bar-clip assembly or magnet arrangements. PMID- 8239413 TI - Localization of nonpalpable masses in patients with breast implants. AB - Nonpalpable nodules in patients with breast implants may represent silicone granulomas, fibrocystic mastopathy, or cancer. We describe a modified technique for needle localization which facilitates the surgical excision of these nodules while minimizing the possibility of rupture. PMID- 8239414 TI - Fine needle aspiration biopsy in patients with augmentation prostheses and a palpable mass. AB - Six patients with augmentation prostheses presented with a firm, painless, breast mass that could not be visualized by mammography. One lesion was demonstrated to be solid by ultrasound, and the remaining sonograms were nondiagnostic. The lesions were indistinguishable from carcinoma, by physical examination. All of the patients had fine needle aspiration biopsy despite close proximity to the implant. The patients all had silicone granulomas related to silicone leakage. Our experience suggests that fine needle aspiration biopsy is a useful technique to evaluate palpable breast masses that are not visualized by mammography in patients with augmentation prostheses. PMID- 8239415 TI - Toe-to-hand transfers in the rehabilitation of frostbite injury. AB - In our experience digitless hands from frostbite injury are typically seen in young men as a result of an episode of intoxication. Such deformities occur bilaterally in 80% of cases and often lead to total loss of prehensile capability. Because of a uniformly high degree of motivation in this group of patients, toe-to-hand transfer carries an excellent prognosis. Microanastomoses in a frostbitten hand are as reliable as in a mechanically injured hand, provided the ulnar artery and its branches are intact. Because amputations occur at the level of the metacarpal heads, the preferred transplant is the combined second and third toe composite. However, a satisfactory thumb post is essential for proper function of the composite. Well-planned sharing of available toe resources is necessary for establishing optimum bilateral hand function, especially in the face of simultaneous toe frostbite. Squandering valuable foot resources bilaterally on behalf of a single hand makes reconstructing the opposite hand more difficult. Economy may be gained if two separate microvascular toe transfers are taken from the same foot or if a single block of toes may be applied to the hand as an opposable unit. Clinical management is described of 25 digitless patients whose frostbitten hands were treated by microsurgical toe transfer. PMID- 8239416 TI - Toe-to-hand transfers in children. AB - One hundred three microvascular toe-to-hand transfers were performed on 66 pediatric patients with congenital and posttraumatic deformities. Techniques for dissection and transfer of individual and composite toes were elaborated, and adjunctive procedures were added when indicated. PMID- 8239417 TI - A comparison of the early antithrombotic effects between low molecular weight heparin and heparin in small arteries following a severe trauma: an experimental study. AB - Twenty-seven rabbits divided into 4 groups (2 control groups) received saline, standard heparin (400 IU both as anti-F Xa and activated partial thromboplastin time per kg of body weight), or low molecular weight heparin (lmwh) (Fragmin 560 IU as anti-F Xa and 140 IU as APTT units per kg of body weight). The figures represent the total dose given over 3 hours. The central arteries of the rabbits' ears were prepared and positioned in clamps. Platelets labeled with 32P were injected. Arteriotomy/intimectomy was performed. After reestablishment of blood flow, arteriotomy bleeding times, platelet accumulation, patency, and weight of thrombotic materials were measured. The bleeding time in the Heparin Group was significantly prolonged compared with its control group. But the bleeding times between the LMWH Group and the control group were not significantly different. The patency rates were increased in both the Heparin Group (100%, p < 0.01) and the LMWH Group (73%, p = 0.078) compared with respective control groups (43-50%). The mean weights of thrombotic material in each artery were significantly lower in the Heparin Group and in the LMWH Group than in their control groups. The mean values of radioactivity in all groups increased up to 15 minutes after the vessels were reperfused. There were no statistical differences between the treated groups and control groups. It was concluded that standard heparin is a very powerful inhibitor of thrombosis during the most crucial hours after reperfusion of severely traumatized small arteries, without significant effects on primary platelet adhesion/aggregation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239418 TI - Transverse rectus abdominis musculocutaneous flaps for lightweights: is a pinch enough? AB - Recent publicity concerning silicone breast implants has fostered patient interest in autogenous breast reconstruction after mastectomy. The transverse rectus abdominus musculocutaneous flap has become the standard for autogenous breast reconstruction. However, very thin patients have not been considered ideal candidates for this procedure due to the paucity of abdominal fat. The use of this procedure in these very thin patients has not been addressed. In patients whose abdominal skin fold thickness was 1 cm or less, we found the procedure to be reliable. However, maximizing the amount of transferred tissue to achieve symmetry necessitates a bipedicled flap. Abdominal fascial closure in these very thin patients is more difficult. Although these patients may not appear to be ideal candidates for the transverse rectus abdominis musculocutaneous flap at first glance, the results have been gratifying and this method of reconstruction is a reliable option. PMID- 8239419 TI - Extensive malignant melanoma of the uvea in childhood: resection and immediate reconstruction with microsurgical and craniofacial techniques. AB - Malignant melanoma is a rare tumor in the pediatric age group. Those that have been reported generally describe cutaneous lesions. In children, malignant melanoma arising in the eye is exceedingly rare. The child we report was born with a uveal tract malignant melanoma with extensive extraocular invasion that was successfully managed with combined craniofacial and microsurgical techniques. This patient demonstrates the difficulty of making the correct diagnosis, the initial reluctance about surgical intervention in a newborn's condition, and the subsequent success possible with standard microsurgical and craniofacial techniques. PMID- 8239420 TI - Double-opposed Z plasty for upper eyelid entropion correction. AB - Recurrent cicatricial entropion of the upper eyelid creates a surgical problem. We present a 53-year-old patient with complicated postsurgical fibrotic and scarred upper eyelid entropion. Double-opposed Z-plasty incisions were used at the anterior approach. The outer Z incision included the skin and the orbicularis oculi muscle underneath, whereas the inner Z incisions included the tarsal plate and conjunctiva, in a mirror-image fashion. Two goals were achieved, i.e., release of the vertical traction vector of the conjunctiva and balance of forces between the anterior and posterior layers of the lid. Symptomatic and clinical improvement was observed, and there was no recurrence 2 years after surgery. PMID- 8239421 TI - Phallic construction in female-to-male transsexuals using a lateral upper arm sensate free flap and a bladder mucosa graft. AB - We devised a technique for phalloplasty in a female-to-male transsexual combining the lateral upper arm free flap with a bladder mucosa graft, to address the ideal requirements of phalloplasty in the best possible way. The surgical techniques are described. Healing of the phallus has been uneventful, but postoperative bladder spasms, meatal stenosis, and an intraurethral valve were encountered. The cosmetic result of the phalloplasty is pleasing. Sensibility is still increasing after a 14-month follow-up. Scarring of the upper arm donor site can be camouflaged easily. Although the actual phalloplasty as described in this article is a one-stage microsurgical procedure that appears to be reproducible, the construction of a phallus meeting all requirements still implies more than one stage. PMID- 8239422 TI - The use of sandpaper in chemical peeling combined with dermabrasion of the face. AB - The combined use of chemical peel and dermabrasion of the face was widely accepted since the first reports of DuPont and Horton. We present a refinement of this combined method by introducing the use of a simple diathermy head cleaner (Surgikos, Inc) as a safer method of face abrasion. This disposable device, available at every surgical theater allows a better control of the abrasive tool, permits a more accurate depth of the abrasion, and reduces the incidence of unexpected complications in this combined technique. PMID- 8239423 TI - Attempted improvement in mammography by inducing a phase transition in silicone gel. AB - An attempt was made to achieve a temporary alteration of the molecular structure (a "phase transition") of silicone gel, the end result of which would be to improve mammography. It was hoped that a phase transition of this type would result in either a reduction in gel volume or increased radiopenetration. Unfortunately, an electrically induced phase transition of silicone gel was not seen. In fact, a temporarily altered molecular orientation (phase transition) was not even possible under the extremes of voltage confirming the fact that silicone is extremely inert. PMID- 8239424 TI - Liberte, egalite, personnalite. PMID- 8239425 TI - Nasal reconstruction with auricular microvascular transplant. AB - A free flap derived from the ascending helix of the ear has been used to reconstruct an anatomically diverse set of defects of the distal nose in 6 patients. Our cadaver injection studies have demonstrated that the blood supply to the auricular flap is via small, consistent branches from the superficial temporal artery. The patients were carefully selected, and most had failed prior attempts at reconstruction. The auricular flaps were used to reconstruct the nasal tip, ala, columella, and sill. The donor vessels were anastomosed either to the facial artery and vein or to vessels in the neck, via vein grafts. The flaps survived in all cases, but all patients required minor subsequent revision. The auricular donor site was closed by rotation and advancement of local tissue. This flap is presented as a surgical option for selected patients with complex defects of the distal nose, where excellent match of color and contour, predictable outcome, and avoidance of central facial donor site are desired. PMID- 8239426 TI - Microvascular reconstruction of nose and ear defects using composite auricular free flaps. AB - Two cases using composite auricular free flaps, based on the superficial temporal vessels, in the reconstruction of full-thickness defects of the nose and ear are reported. This composite free flap can be based on either the superficial temporal or the posterior auricular vessels depending on the defect to be reconstructed. The superficial temporal vessels can also be used in reversed flow to obtain a vascular pedicle of sufficient length for microvascular anastomosis. The anatomical vascular features of the flap make it possible to reconstruct various facial defects with freedom of design. PMID- 8239427 TI - Risks and benefits of using Marlex mesh in chest wall reconstruction. AB - The records of 101 patients who had undergone chest wall reconstruction both with and without stabilization of the chest wall by Marlex mesh were reviewed to see if Marlex reduced ventilator dependence and hospital stay, and to determine whether the use of Marlex was associated with any increased risk of infection. In 40 patients in whom Marlex was used, the mean number of days on postoperative ventilator support was 0.8, and mean hospital stay was 9.7 days. In 61 patients in whom Marlex was not used, the mean number of days on ventilator support was 4.9, and mean hospital stay was 17.5 days. These differences were statistically significant (p = 0.03, p = 0.006). Two patients in the Marlex-stabilized group (5%) developed wound infections, but these were preceded by ischemic necrosis of overlying flaps. None of the patients without Marlex developed wound infections. That difference was not statistically significant. We conclude that the use of Marlex in chest wall reconstruction does not significantly increase the risk of wound infection, provided that overlying tissues are properly vascularized and remain viable, and that synthetic mesh does improve chest wall stability and reduce ventilator dependence and overall hospital stay. PMID- 8239428 TI - The predictive value of the laser Doppler flowmeter for postoperative microvascular monitoring. AB - Reliable postoperative monitoring in microvascular surgery is necessary to improve the low success rate of reexploration after vascular compromise. The use of laser Doppler flowmetry has been evaluated in this study. From November 1985 to January 1988, 79 microvascular operations were monitored postoperatively. These consisted of 45 replants and revascularizations in 34 patients, as well as 34 free vascularized tissue transfers. In the replant and revascularization group, a statistically significant difference in laser Doppler flowmetry readings has been demonstrated between well-vascularized and circulatory impaired patients (p < 0.001). In this study a reliable critical (alarm) value could be defined for replants and revascularizations with a sensitivity of 93% and a specificity of 94%. This critical alarm value can be adjusted according to the individual surgeon's attitude toward reexploration. Similar laser Doppler flowmetry characteristics were seen in patients with free vascularized tissue transfers; however, the numbers in this part of the study were too small to define reliable critical alarm values. The laser Doppler flowmeter is recommended for the postoperative evaluation of tissue after microvascular anastomoses, as it indicates vascular occlusion at an early stage where reexploration is worthwhile. PMID- 8239429 TI - Tensor fascia lata V-Y retroposition myocutaneous flap: clinical experience. AB - A modification of the standard tensor fascia lata myocutaneous flap, namely, the tensor fascia lata V-Y retroposition flap, is a reliable, durable, efficient, well-tolerated, and technically straightforward procedure. Indications include trochanteric pressure ulcers, nonhealing, failed hip arthroplasties, and soft tissue defects after regional tumor excisions. In 70 TFL V-Y retroposition flap reconstructions performed, the overall postoperative complication rate was 23%, with 4 total flap failures. Sixty-three percent of complications were treated with local wound care, thus avoiding repeat surgery. Rerotation of this flap is not only possible, but also just as reliable as the initial procedure. The tensor fascia lata retroposition flap offers advantages over many of the other tensor fascia lata flap variations. PMID- 8239430 TI - The amplified sliding flap. AB - During recent years, sliding flaps have had great popularity owing to their advantages in terms of reliability, ease of execution, and good morphofunctional results. The limits of these flaps are related to the capacity of excursion in the advancement. To improve these limits, we describe in this article a personal modification of the flap that extends the possibilities of coverage in the distal areas, reducing the tensile distress. We report our experience with 19 patients, treated with this modified flap in various areas of the body, with good results and without noticeable complications. PMID- 8239431 TI - Long-term follow-up of hand function after digit replantation. AB - A long-term follow-up study of 26 patients with 28 replanted digits was performed with respect to circulatory, sensory, and motor function. Peripheral circulation in the replanted digits was comparable with corresponding uninjured digits on the opposite hand. Excellent sensory recovery was observed in the territories supplied by 30 of the 36 repaired digital nerves. Best functional results were noted with the thumb and with fingers amputated through the middle phalanx. By objective criteria, 21 of the 28 replanted digits demonstrated satisfactory function. Subjectively, however, only 1 patient was dissatisfied with the outcome. PMID- 8239432 TI - Flap transfer for complications of major limb replantation. AB - Our experience in the management of necrotizing and suppurating wound complications after major upper extremity replantation is presented. Seven patients underwent wound debridement and coverage with flaps. Of these, 6 were reconstructed with latissimus dorsi flaps and 1 with the scapular flap. In 3 of these patients, concomitant hemorrhage (2 patients) or thrombosis (1 patient) was simultaneously corrected by vein grafting. In all cases there was survival of both the flap and the replanted extremity. PMID- 8239433 TI - Application of basic fibroblast growth factor may reverse diabetic wound healing impairment. AB - This study examines the effect of application of basic fibroblast growth factor (bFGF) on the wound healing impairment seen in streptozotocin-induced diabetic (SD) rats compared with control nondiabetic (NL) animals. Under general anesthesia, a 6-cm dorsal incision was made through the skin and panniculus carnosus. Both the NL and SD wounds were injected with 0.1 ml of one of the following three solutions: saline, vehicle, and 10 micrograms of bFGF. The wounds were closed with interrupted sutures of 4-0 nylon. The animals were returned to their cages and sacrificed at 7, 10, 14, or 21 days later. Breaking strength of the wound was analyzed by using an Instron Tensiometer 4201 to assay for relative collagen maturation. Data among groups were compared using an analysis of variance. At 7 and 10 days, all NL wounds were stronger than all SD wounds. By 14 days there was no statistically significant difference in breaking strength between the bFGF-SD wounds and all subgroups of NL wounds. The bFGF-SD subgroup was statistically significantly stronger than the saline-SD (p < 0.02) and vehicle-SD (p < 0.01) wounds. At 21 days the bFGF-SD wounds were statistically significantly stronger than the vehicle-SD wounds to a confidence level of p < 0.001. These findings indicate that application of bFGF may reverse the impairment seen in diabetic wound healing. PMID- 8239434 TI - Duration of contact time alters cutaneous pressure threshold measurements. AB - Semmes-Weinstein monofilaments (SWMs) are standard measures used to assess hand sensibility. The purpose of this study was to evaluate the effect of varying contact time on the measurement of cutaneous pressure thresholds. Thirty-two healthy adult subjects were evaluated with a 20-filament SWM test kit. The index finger of the dominant hand was assessed with the following four distinct monofilament contact times: 0.5, 1.5, 5.0, and 30.0 seconds. With increased duration of monofilament contact time, there was a significant increase in the cutaneous pressure threshold measures (p < 0.05). That is, to perceive a stimulus for a longer period of time, a heavier filament was required. With increasing age, significantly higher pressure thresholds were obtained particularly for the sustained contact time (> 5.0 seconds). Pressure thresholds vary significantly with SWM contact time, emphasizing the importance of maintaining a consistent SWM application protocol. PMID- 8239435 TI - A cultured skin substitute composed of fibroblasts and keratinocytes with a collagen matrix: preliminary results of clinical trials. AB - The cultured skin substitute was created through successive cultivation of fibroblasts and keratinocytes that were combined within a collagen matrix. This collagen matrix was composed of a collagen spongy sheet and a collagen gel. The collagen spongy sheet was designed to produce a honeycomb structure having many holes in which all holes through the sheet were filled with collagen gel. This specific structure thereby allows for the nourishment of the cultured keratinocytes on the surface of the matrix when placed on the graft bed. In this study, autologous cultured skin substitute was applied to a 51-year-old man who had sustained a burn injury. Three sheets of the cultured skin substitute (6 x 9.5 cm) were grafted onto the full-thickness excised wound in the right anterior chest wall. One week after grafting most of the matrix disappeared and stratified keratinocytes were seen to have firmly attached to the underlying tissue. Five weeks after grafting a cornified epidermal layer was seen. Ten months after grafting a mature epidermis and a well-differentiated papillary and reticular dermis replacement were observed. The physical properties and color of this grafted area resemble those of normal skin. In the second test case, autologous cultured skin substitute was applied to a 30-year-old man with a scar remaining after tattoo removal. Eight sheets of the cultured skin substitute (10 x 18 cm) were applied on an excised wound (thickness, 0.02-0.025 in.) of both the fore- and upper arms. The histological appearance of a biopsied skin specimen from the grafted area at 3 months after grafting showed a mature epidermis and a well differentiated reticular dermis replacement. The regenerated skin at 14 months after grafting showed an excellent result. PMID- 8239436 TI - The rat rectus abdominis myocutaneous flap: a true myocutaneous flap model. AB - Investigations of experimental flaps in lower-order animals have concluded that the cutaneous vascular anatomy of "loose-skinned" animals is not analogous to that of humans. This is particularly relevant with respect to myocutaneous flaps. The interpretation of results of studies in lower-order animals raises questions as to the applicability of these data to the human condition. OBJECTIVES: To establish a true myocutaneous flap model in the rat, to define the microvascular anatomy and the cutaneous vascular territory of the rat rectus abdominis myocutaneous flap, and to determine its suitability for additional investigative study of myocutaneous flaps. METHODS: Forty-two Sprague-Dawley rats weighing 300 to 400 gm were divided into the following six experimental groups: Group 1 (microvascular anatomy, n = 5), microfil injection and microdissection; Group 2 (n = 5), India ink injection of an isolated superior or inferior epigastric artery pedicle; Group 3, flap control (n = 6); Group 4, superior epigastric (n = 17); Group 5, deep inferior epigastric (n = 16); and Group 6, bipedicle deep inferior epigastric (n = 3). Rectus abdominis myocutaneous flaps based on a single superior or inferior vascular pedicle, or on a double inferior vascular pedicle with an overlying cutaneous island extending from the costal margin to the pubis and anterior to the anterior axillary line, were raised and replaced. Control flap vascular pedicles were transected. Assessment of viability was performed at 7 days using two-dimensional planimetry, tetrazolium, and hematoxylin and eosin stain. RESULTS: Injection studies revealed large (0.1 mm), direct, perforating vessels from the rectus abdominis muscle to the overlying cutaneous island. The cutaneous territory was defined by isolated pedicle india ink injection extended over the vertical muscle axis to the anterior axillary line and for a variable distance across the midline. All controls flaps were nonviable at 7 days. Superiorly based single-pedicle, and double-pedicle inferiorly based, flaps had 97% (SD, 0.03) survival of the cutaneous island. Single inferior pedicle flaps had 77% (SD, 0.07) survival of the cutaneous island with necrosis of the contralateral distal skin island. All flaps had 100% muscle survival. CONCLUSION: The rat rectus abdominis muscle and overlying cutaneous island constitutes a true myocutaneous territory. This has not been described in a low-order laboratory animal. The flap can be based on single or double, superior or inferior, epigastric vessels with predictable flap survival. The superiorly based and double pedicle flap demonstrates nearly complete viability. This model should serve as a useful investigational tool in the laboratory study of myocutaneous flaps. PMID- 8239437 TI - Epidermal cell viability in rat skin preserved at 4 degrees C. AB - To determine the longest storage period for the clinical application of epidermal cells and a better method of preservation at 4 degrees C, we stored skin in various fluids and assessed the changes in epidermal cell viability. The time course of changes in the viability of epidermal cells from skin in various preservation fluids was similar to the results obtained with whole skin in previous studies. The best preservation fluid was found to be RPMI 1640 medium, and the best method of preservation was immersion in RPMI 1640 at 4 degrees C with a medium change every 3 days. PMID- 8239438 TI - A combination surgical method for preaxial polydactyly of the foot. AB - A combination procedure was performed on the medial two toes in a 6-month-old child with preaxial polydactyly of the left foot. The first toe had the appearance of a cosmetically intact great toe, but the second toe had better function. A method to combine the advantages of these two toes was therefore applied, with favorable results seen at 1.5 years after surgery. PMID- 8239439 TI - Necrotizing periorbital cellulitis. AB - We report traumatic necrotizing periorbital cellulitis attributed to group A beta hemolytic streptococci in a 4-year-old child. The infection was successfully treated via surgical cleansing, drainage, and grafting. The virulence of this organism requires an aggressive approach to the patient with periorbital cellulitis, which is refractory to intravenous antibiotics. Early treatment may limit extensive eyelid necrosis, the resultant secondary deformity, and the need for multiple reconstructive procedures. PMID- 8239440 TI - Scrotal construction by expansion of labia majora in biological female transsexuals. AB - Construction of male genitalia in biological female transsexual patients is problematic. We report 3 transsexuals with scrotal construction by tissue expansion of the labia majora. In each patient, tissue expanders were inserted into the labia majora several months before free flap phalloplasty. In 2 patients, the tissue expanders were removed, and testicular implants were placed into the labial pockets. In the other patient, the tissue expanders were left in place. Because of the close embryological relationship between the labia majora and the scrotum, the neoscrotum appears natural in color match, hair pattern, and skin texture. The anatomical position of the labia majora produces a scrotum properly located in relation to surrounding anatomical structures. Excellent aesthetic results can be achieved with this procedure. Based on our experience, we advocate this procedure for scrotal construction in biological female transsexual patients. PMID- 8239441 TI - Hand metastasis from renal cell carcinoma with no bone involvement. AB - Hand metastases account for 0.1% of all metastases and most of them involve primarily the bones. A case of a metastasis from renal cell carcinoma to the distal segment of the little finger with no bone involvement is described. In view of the fact that the kidneys are the origin of only 10% of hand metastases, such a case is extremely rare. PMID- 8239442 TI - Re: Use of tissue expanders in reconstruction after excision of multiple large silicone granulomas. PMID- 8239443 TI - Re: Nipple-areola reconstruction with intradermal tattoo and double-opposing pennant flaps. PMID- 8239444 TI - Re: Reconstruction of a medial canthus defect with a myocutaneous flap. PMID- 8239445 TI - Re: Myonecrosis as a sequela of calf implants. PMID- 8239446 TI - The transverse gracilis musculocutaneous flap. PMID- 8239447 TI - John Hunter and his painters. PMID- 8239448 TI - Research in higher surgical training--the West Midlands view. AB - A questionnaire on the role of research in higher surgical training was posted to all in-post NHS Consultants (n = 96) and Higher Surgical Trainees (n = 42) in the West Midlands Region. Replies were received from 80 consultants (83 per cent) and 37 trainees (88 per cent). Over 95 per cent of responders had undertaken some research activity previously. The vast majority of consultants (90 per cent) and trainees (95 per cent) thought that their research period had been beneficial; most would undertake research again even if not essential for higher surgical training (consultants 80 per cent, trainees 70 per cent). Research was felt to be essential for all trainees by 50 per cent of respondents in both groups; 78 per cent of consultants and 86 per cent of trainees also thought that all potential academics should obtain a degree by thesis. Most of the study participants thought that the appropriate time for research activity was as a post-FRCS registrar (consultants 72 per cent, trainees 80 per cent); 80 per cent of consultants and 67 per cent of trainees felt that this research period should be funded by the NHS. A planned, supervised and funded one-year period of research was favoured by the majority of consultants (54 per cent) and trainees (73 per cent) for non-academic general surgical trainees in the future. PMID- 8239449 TI - An explanation of the role of individual performance review in surgical training. AB - The consultant's success depends upon his/her staff. If the consultant is to provide a more efficient service the staff must be encouraged to develop and supplement their own skills. Individual Performance Review (IPR) is a technique that has been introduced to develop the managerial skills of NHS staff. However, its considerable benefits have not yet been extensively applied in clinical specialties. This article describes the current role of IPR and appraisal interviews in the process of training surgical house officers and senior house officers in a district hospital. PMID- 8239450 TI - What is a reasonable orthopaedic surgical workload? An analysis of elective and trauma workloads using two different complexity scoring systems. AB - A 1-year prospective study of all surgical activity was undertaken by the Orthopaedic Unit using two weighting systems for operative complexity: the Intermediate Equivalent (IE) and the Operative Complexity Score (OCS). The objectives were to define the difference between caseload and workload, and to formulate a reasonable workload for an operating session and a reasonable annual workload for a consultant team. In both elective and trauma surgery the BUPA complex, major+ and major operations represented 50 per cent of the caseload but accounted for approximately 68 per cent of the workload. The BUPA minor and intermediate procedures representing the remaining 50 per cent of the caseload accounted for only 32 per cent of the workload. The number of hours required to undertake all the complex, major+ and major operations was approximately twice that required for the number of minor and intermediate procedures. Despite the marked variation in case mix, the workload achieved by the six teams in the same number of sessions was remarkably similar. On the basis of two consultant and one registrar elective lists per week, a total of 705 IEs (1795 OCS units) could be undertaken by each team in 1 year. An average of 458 IEs (1086 OCS units) of trauma surgery were undertaken by each team. Caseload is a poor performance indicator of surgical activity. This study supports the concept of 3.5-4.0 IEs as reasonable workload for an orthopaedic elective operating session, and 900 IEs (2200 OCS units) as a reasonable annual workload for an orthopaedic team. PMID- 8239451 TI - [Meniscal lesions]. AB - The polymorphism and richness of arthrographic imaging and MRI of meniscal lesions require a perfect understanding of the anatomy of the menisci and the mechanisms of production of the lesions. Arthrography remains the reference diagnostic investigation with a good sensitivity and excellent spatial resolution. However, it is an invasive and operator-dependent technique. MRI of the knee, a non-invasive and independent method, allows the diagnosis of the majority of meniscal lesions. By using specific surface coils and appropriate sequences, MRI constitutes complete imaging technique allowing examination of all anatomical structures of the knee in all planes. Once MRI becomes more widely available, it should replace arthrography in the majority of clinical situations. The indications for arthrography will then be confined to contraindications of MRI. PMID- 8239452 TI - [The femoropatellar joint]. AB - Anomalies of the femoropatellar joint are frequent due to the vulnerable anatomical situation of the patella. Any knee abnormality, constitutional as well as acquired, may compromise the precarious dynamic balance existing between external and internal forces applied to the patella. PMID- 8239453 TI - [The role of MRI of the knee]. AB - Imaging of the knee is one of the main indications of MRI in musculo-skeletal diseases. Some of the most recent papers dealing with MR imaging of the knee are discussed in this review article. Abnormal conditions reviewed are meniscal injuries including those occurring in previously operated menisci, osteochondral fractures, osteochondritis dissecans and rheumatoid arthritis. PMID- 8239454 TI - [Ligaments of the knee]. AB - The authors study the various ligaments of the knee and, for each one, the anatomy, mechanisms of injury, clinical diagnosis, indications for imaging (plain x-rays, arthroscopy, MRI). The anterior cruciate ligament and posterior cruciate ligament are best visualised by MRI. The tibial collateral ligament and fibular collateral ligament are also well visualised by MRI. The value of the posteromedial and posterolateral angle points, stabilising functional structures, is emphasised together with the important diagnostic distinction between isolated ligamentous lesions and associated lesions. MRI constitutes the best imaging technique for assessment of the lesions. It completes the clinical examination and should be performed early. PMID- 8239455 TI - [Course of recurrent dislocation of the patella, patellar syndrome without dislocation and femoropatellar osteoarthritis]. AB - The frequency of recurrent dislocation of the patella is increased by the young age of onset, the presence of patella alta and permanent subluxation of the patella. The natural history of the patellar syndrome appears to be fairly favourable in everyday activities, but discomfort is often marked in the intermediate term in sports activities. Isolated femoropatellar osteoarthritis rarely causes major functional impairment of gait. Medial or lateral femorotibial osteoarthritis occurs secondarily. Painful cases often present with subchondral anomalies of the lateral patellar facet, lateral tibial osteophytosis and patella alta. PMID- 8239456 TI - [Spontaneous course of femorotibial osteoarthritis]. AB - Medial femorotibial osteoarthritis (MFTO) is characterised by progressive deterioration resulting, in 6 out of 10 cases, in an inability to walk after 10 to 15 years. At the surgical stage, AP films show subchondral bone wear on the edge of the medial tibial plateau with lateral tibial dislocation and lateral films show obliqueness of the medial tibial plateau possibly exceeding 10 degrees and accompanied by anterior subluxation of the tibial or even rupture of the anterior cruciate ligament. Lateral femorotibial osteoarthritis (LFTO), more frequent in women, occurs an average of 7 years later than MFTO, at an average age of 70 years. Instability is often severe. 72% of cases develop clinical valgus deformity of 10 degrees to 20 degrees after an average of 13 years. Recurvatum is also possible. The combination of femoropatellar osteoarthritis, generally lateral, is observed in 23 to 41% of cases, depending on the author, while osteoarthritis of the hip is observed in 10% of cases and chondrocalcinosis is present in 20% of cases. These figures must be interpreted in relation to the usual ageing of the subjects. PMID- 8239457 TI - [Surgical treatment of non-osteoarthritic femoropatellar diseases]. AB - Patellar instability, associated with varying degrees of pain, is the major symptom of non-osteoarthritic femoropatellar diseases. This patellar instability is due to either mechanical (femoropatellar "derailment") or pain (due to chondropathy) disruption of quadriceps function. The result of surgical treatment of painful femoropatellar instability, regardless of its type, is governed by the same postoperative factors. A good result can only be obtained when the postoperative TA-GT lies within a certain range of the ideal value which varies according to the trochlear angle; the quality of the result essentially depends on the postoperative height of the patella. Preoperative radiological measurement of the trochlear angle on femoropatellar views in 30 degrees of flexion, of the TA-GT angle in 30 degrees of flexion and the height of the patella is therefore necessary to select and modulate the therapeutic procedures designed to treat non osteoarthritic femoropatellar diseases. PMID- 8239458 TI - [Outcome of femoral osteochondritis dissecans of the knee]. AB - Osteochondritis dissecans is a radiological entity, corresponding to a juxta articular bone fragment separated from the rest of the epiphysis by a radiolucent zone. Diagnosed prior to closure of the femoral growth plate, the abnormal image generally disappears without leaving any sequelae. No aggressive treatment is therefore indicated. The course of osteochondritis detected after closure of the femoral growth plate is different: the sequestrum always evolves spontaneously towards separation from the epiphysis to become an intra-articular foreign body. Osteochondritis in adults also predisposes to early osteoarthritis. Fixation of the sequestrum while it is still in place clearly protects the knee form osteoarthritic deterioration and should be proposed without hesitation whenever a periosteochondritic fissure is observed. PMID- 8239459 TI - [Occult fractures of the internal tibial plateau]. AB - Insufficiency bone fracture of the medial tibial plateau occurs in elderly patient. Pain and disability are important. Physical examination and initial X Ray are note contributive. Insufficiency fracture appeared on bone scan as localised hyperfixation and as a slow intensity image of the medial tibial plateau on MR Imaging. Outcome is favorable with discharge. PMID- 8239460 TI - [Correlation and discordance between arthrography and arthroscopy of the knee]. AB - We retrospectively compared two series of 50 consecutive arthroscopies, performed in 1980 and in 1990. The age and sex data of the group were essentially the same. The number of arthrographies performed prior to arthroscopy varied only slightly (64% versus 58%), as did the indications, dominated by uncertain clinical findings (58%). One quarter of the procedures constituted useless indications in an obvious clinical context. These 2 groups were studied separately, cumulatively and according to the diagnoses. Analysis showed a better arthrography-arthroscopy correlation in 1990. The arthrographic technique and the technician himself appear to be important factors. Progress in the technique has increased the quality of imaging of certain anatomical structures. This good overall concordance was also observed for analysis of the diagnoses. We therefore conclude that the current indications for arthrography prior to arthroscopy are any uncertain diagnosis after a thorough clinical examination, when meniscal suture is considered, lateral meniscal cyst and a medicolegal context. PMID- 8239461 TI - [Technique of MRI of the knee. Principle and procedure]. PMID- 8239462 TI - [Three-dimensional MRI of the knee]. AB - Three-dimensional gradient echo T2-weighted sequences have a number of advantages over spin echo T2-weighted sequences (or even 2D gradient echo T2-weighted sequences) for assessment of the knee. They allow a multidimensional analysis based on a single acquisition sequence usually obtained in the sagittal plane. Image reconstructions can be performed secondarily in the coronal, axial and oblique planes, particularly along the specific path of the anterior cruciate ligament. By providing ultrathin serial sections, decreasing the partial volume effect, small lesions, such as cartilaginous fissures or flaps and radial meniscal lesions can be detected in the axial plane, for example. This advantage, combined with the marked sensitivity of gradient echo sequences to alterations in the tissue water content, allows the detection of partial tendon ruptures. The reduction of the partial volume effect and chemical shift artefact probably participate in the capacity of these sequences to visualise the two surfaces of the cartilage of the femorotibial joint. Flow artefacts are less of a problem than with 2D imaging, which eliminates the need for techniques such as saturation of the vascular signal or cardiac gating. A disadvantage of these gradient echo sequences (3D or 2D) is their sensitivity to the presence of metallic material, limiting their application in operated knees. PMID- 8239463 TI - [Management of a suspected lesion of a cruciate ligament]. AB - After classifying and describing the epidemiology of various traumatic ligament lesions of the knee: "isolated rupture", "triads" and "pentads", the author describes their mechanism and clinical presentation. Plain x-rays and stress x rays, possibly under anaesthesia, are necessary to establish the diagnosis of the lesions. In 1991, in the absence of triads, pentads or radiological bone avulsion, they are followed by diagnostic arthroscopy to determine the partial or complete nature of the cruciate ligament rupture; stress films and arthroscopy under anaesthesia can be advantageously replaced by high performance MRI in the months to come. PMID- 8239464 TI - [Radiological assessment before femoral and tibial osteotomies]. AB - After recalling the good results of femoral and tibial osteotomies for lateral osteoarthritis of the knee before the age of 70 years, the author describes the usual techniques and defines the indications. He then defines the preoperative x rays essential for such procedures: AP and lateral films of the knee in 15 degrees of flexion, AP in forced varus and valgus with 15 degrees of flexion, femoropatellar view with 30 degrees of flexion with particular attention to "stepping" gonometry, which appears to be the most physiological approach. PMID- 8239465 TI - [Radiological examination of knee prostheses]. AB - After defining the respective indications for the various knee prostheses: single compartment, three-compartment sliding and hinge prostheses, essentially based on the preoperative radiological assessment, the authors emphasise the value of such films in operative planning. They then describe the radiological criteria of correct implantation of the three different types of prosthesis. Lastly, they stress the value of regular radiological follow-up of these arthroplasties looking for possible complications such as instability, loosening, wear and infection, which may alter the patient's osteoligamentous capital and his functional prognosis. PMID- 8239466 TI - [Indentation sign in the sylvian sulcus and the arterial origin of an acute subdural hematoma. Contribution of computerized tomography]. AB - Thirty-two traumatic subdural haematomas were operated between 1984 and 1990. In 16 cases (50%), we encountered, after removal of the heamorrhage, a bleeding from a cortical artery at the lateral sulcus (middle cerebral artery). Four mechanisms of arterial bleeding have been reported: a) rupture of a cortico-dural bridging artery, b) "fire-hose" rupture, c) rupture at the level of an arachnoid attachment and d) rupture at the level of a dural adhesion. The characteristic finding of such haematomas on computerized tomography (CT-scan) was an indentation towards the lateral sulcus. This was called the "double brackets sign". In seventy-nine percent of subdural haematomas of arterial origin, the "double brackets sign" was demonstrated on CT-scan on at least two slices (9 mm). In 9% of subdural haematomas of non-arterial origin, this sign was absent or present in only one CT image. The characteristic "double brackets sign", when present on 3 or 4 CT-scan images, seems to have a very high specificity for an arterial origin of the haematoma. PMID- 8239467 TI - [Computerized tomography and preoperative evaluation of cancers of the esophagus. Apropos of 54 anatomo-radiological comparisons]. AB - The authors report their experience in the study of esophageal carcinoma before surgery by CT. Computed tomography is essential to establish the local extension of the tumor process and thus to allow a better therapeutic management. PMID- 8239468 TI - [Percutaneous drainage of splenic fluid accumulations. Apropos of 4 cases]. AB - Four patients with solitary or multiple splenic fluid collections were treated by ultrasound guided percutaneous needle aspiration and drainage. There was one unilocular abscess, three infected hematomas, one large hematoma and a complex lesion with locular and perisplenic involvement. Percutaneous drainage and antibiotics were curative in one patient. Splenectomy was performed because of persistent fever and drainage of pus after 15 days in one case and recurrence of hematoma in an other case. Solitary fluid collection can be effectively treated by ultrasound or computed tomography guided percutaneous drainage. Hematoma represents a cause of failure of percutaneous drainage. PMID- 8239469 TI - [Ultrasonography in the diagnosis of liver abscesses. Apropos of 32 cases]. AB - A retrospective series of 32 cases of hepatic abscess was collected in the radiology department of Ibn Roch University Hospital (Casablanca), over a period of 43 months (May 1989-September 1992). Among these 32 cases, 16 were amebic (based on the dysenteric syndrome and especially positive amebic serology) and 16 had pyogenic causes. All patients underwent ultrasound examinations. Only two cases had CT scan. The confirmation of the hepatic abscess was made by ultrasound guided percutaneous aspiration in 30 cases and laparotomy in 2 cases. The right lobe was concerned in all cases. The pyogenic abscesses were multiple in 62.5% cases, while the amebic abscesses were double in only one case. The amebic abscesses were larger than the pyogenic ones. The average volume was respectively 10.5 cm and 7 cm. Three ultrasound aspects were found. Hypoechogenic aspect was the most frequent (76% of amebic abscesses and 61% of pyogenic abscesses). The heterogenic aspect was found in 21% of amebic abscesses and in 36% of pyogenic abscesses. The anechogenic aspect was found in only one case of pyogenic abscess. A light and regular wall was found in 70% of amebic abscesses and in 51% of pyogenic abscesses. Referring to the literature, the various ultrasound signs of orientation towards an amebic or pyogenic etiology were found in this series, but we emphasize the very important role of ultrasound-guided percutaneous aspiration and amebic serology for the etiological diagnosis of hepatic abscess. PMID- 8239470 TI - [Aseptic osteomyelitis]. AB - Based on a series of eight personal cases and a review of the literature, this study tries to define the diagnostic elements of aseptic osteomyelitis. Although sternocostoclavicular sites are immediately suggestive of the diagnosis, lesions of the pelvis and spine and long bones of the limbs are particularly difficult to diagnose. Inflammatory type pain occurred in episodesover a number of years and responded to NSAIs and possibly diphosphonates. Laboratory abnormalities were usually confined to a raised ESR, but alterations of the IgA levels, similar to those observed in ankylosing spondylitis were observed in four cases. Hyperostosis occurred late in the course, preceded by signs of osteomyelitis, periosteitis or enthesitis. Histology does not provide any formal conclusions. The most suggestive lesions are those of nonspecific aseptic osteomyelitis, followed by a Paget-like appearance. The interpretation of these findings may be complicated by the presence of eosinophils, giant cells or mast cells. Two elements are very valuable for establishing the diagnosis: the presence of infraradiological anterior thoracic increased uptake on bone scan and the presence of a skin disorder, either severe acne or, more especially, palmoplantar pustulosis. PMID- 8239471 TI - [Superior vena cava thrombosis. Radiological aspects. Apropos of 28 cases]. AB - The authors report twenty-eight cases of superior vena cava thrombosis. They try to establish a correlation between the radiological features of thrombosis and the etiology. The etiology was Behcet's disease in 64% of cases, lung cancers in 14%, the mediastinal tuberculosis in 14%, systemic lupus erythematosus in 3.5% and undetermined origin in 7%. A normal meiastinal appearance slight widening of the superior mediastinum is noted in Behcet's disease, systemic lupus erythematosus and tuberculous mediastinal fibrosis. Superior vena cava thrombosis is extensive in Behcet's disease involving the subclavian vein and the azygos vein. An obvious widening of the mediastin is noted in lung cancers and tuberculous mediastinal lymphadenopathy. PMID- 8239472 TI - [Pulmonary artery pseudoaneurysm caused by a Swan-Ganz catheter and treated by embolization]. AB - The frequency of life-threatening complications induced by Swan-Ganz catheter is estimated at 2% to 17%. Pseudoaneurysm formation secondary to traumatic injury of the pulmonary artery is rare (0.66% to 2%), but carries a high risk of hemorrhagie. Recognition of possible lesions should be based on clinical data and predisposing factors. Diagnostic and therapeutic investigations should be rapidly performed. Embolization is the best treatment giving very good results. Should acute hemorrhage occur in the operating room, emergency vascular occlusion can be carried out by the use of the Swan-Ganz catheter. PMID- 8239473 TI - [Intrarenal migration of two fractured struts of a vena cava filter]. AB - Filter complications are quite common. The most frequent are distal or proximal migration, angulation, vena cava thrombosis or perforation. We report one case of filter struts fracture and their migration in the right kidney. This complication could require a surgical procedure and demonstrates the importance of filter follow-up with regular abdominal plain X-rays and possibly CT scan and/or cavography. PMID- 8239474 TI - [Marchiafava-Bignami's disease with a favourable course. Apropos of a case]. AB - A resolving form of Marchiafava-Bignami disease is reported. This patient with a history of chronic alcoholism was hospitalised for a recent demential syndrome. Clinical examination did not show any signs of interhemispheric disconnection LP and EEG were normal. CT examination showed a low density area in the corpus callosum without any other anomaly. MRI examination confirmed the cystic area of the corpus callosum. The Marchiafava-Bignami disease was easily suggested on the basis imaging and clinical signs. Interhemispheric disconnection is not always found so MRI and CT examinations seem to be very useful tools for early diagnosis of Marchiafava-Bignami in the same way as neuropsychological tests. PMID- 8239475 TI - [Agenesis of the right lobe of the liver. Apropos of a case]. AB - The authors report a case of agenesis of the right lobe of the liver, only thirty two cases of which have been reported in the literature since 1870. The differential diagnosis consists of advanced cirrhosis, portal hypertension, neoplastic infiltration of the porta hepatis, gallstones or surgical resection. Agenesis is due to arrest of hepatic development during foetal life. It is generally symptomatic, but may sometimes present in the form of biliary type pain in the right hypochondrium or portal hypertension and may be associated with other anomalies (retrohepatic gallbladder, common bile duct cyst, aplasia of part of the diaphragm, intrathoracic kidney). PMID- 8239476 TI - Mucinous cystadenocarcinoma of the pancreas with liver metastases. An unusual presentation of a rare tumor. AB - We present a case of mucinous cystadenocarcinoma of the pancreas with liver metastases. Initially, the patient presented as pancreatitis and pseudocyst which was marsupialised into the stomach in 1983. Eight years later, a tumor with liver metastases was detected. The liver metastases mimicked multiple abscesses. Over a period of forty-four days, CT features changed considerably and there was explosive enlargement of the lesions in the postoperative period. The reason for this dramatic increase in size was the anaplastic component of the tumor. To our knowledge, no such case has been reported in literature. PMID- 8239477 TI - [Amebic brain abscess. Apropos of a case]. AB - We report a case of double primary amoebic brain abscess in a 22 year old man, with signs of intracranial hypertension and deficit. Computed tomography found two hypodense lesions, not suggestive of pyogenic abscesses. Aspiration of one of the two abscesses, revealed the organism, and the two abscesses regressed in response to medical treatment. The authors insist on the rarity of these abscesses, their gravity, and the difficulty of the diagnosis. PMID- 8239478 TI - Pseudotumor cerebri: a review. AB - Pseudotumor cerebri (PTC) is a syndrome seen more frequently in young, obese women defined by increased intracranial pressure without obstruction of the ventricular system. Normal neurodiagnostic studies, normal cerebrospinal fluid (CSF) contents, papilledema in one or both eyes, and normal mental status should also be recognized. Although the disorder has been associated with drugs, endocrinopathies, compromised nutritional status and other disease states, no underlying disorder is uniformly present. PMID- 8239479 TI - Euglobulin lysis times: an update. AB - There are several clot lysis tests available to assess the presence of increased fibrinolysis in a clinical situation. These include whole blood, plasma, and "eugloblin" lysis tests. The euglobulin lysis test (ELT) is actually a modified plasma clot lysis time, which detects increased plasminogen activation and subsequent fibrinolysis. It does not detect the activity of antiplasmins. The purpose of this study was to determine ELT for a group of 25 ambulatory normals. There were 10 males and 15 females, ranging in age from 25 to 56 years. Citrate anticoagulated samples were obtained immediately after application of a tourniquet and 10 minutes after application of a sphygmomanometer inflated to midway between systolic and diastolic pressure. The citrated plasma was precipitated with cold acetic acid. The precipitate contained fibrinogen, plasminogen, and plasminogen activators, with fibrinolytic inhibitors theoretically removed. The precipitate was redissolved and the euglobulin solution was clotted with thrombin. The clotted sample was then incubated at 37 degrees C and was observed at 10 minute intervals for clot lysis. RESULTS: The reference range of onset of fibrinolysis was from 1.25 to 12 hours, with a mean of 3.78 hours +/- 2.45 hours (1 S.D.). Venostasis (use of the sphygmomanometer) resulted in an ELT that was shorter than the non-venostasis specimens in 11 of 25 specimens. This is thought to be secondary to in vivo activation of plasminogen. CONCLUSION: The average euglobulin lysis time in 25 ambulatory normals is approximately 3.5 hours. The ELTs vary over a broad range (1.25 to 12.0 hours) in normal ambulatory volunteers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239480 TI - Lipid peroxidation in platelet concentrates: effects of irradiation and metal chelators. AB - Increased red cell lipid peroxidation (LP) occurs in the presence of ionized iron, as well as with various heme moieties and other transition metal ions. In stored erythrocytes, the degree of LP, as determined by plasma malondialdehyde measurements, can be significantly reduced by the addition of various metal chelating agents. The current study also showed significant reductions in LP in platelet concentrates, compared with controls, following the addition of the metal chelators ethylenediaminetetraacetic acid (EDTA; P < 0.001), diethylenetriaminepentaacetic acid (DTPA; P < 0.001), and deferoxamine mesylate (DM; P < 0.01). In addition, compared with non-irradiated controls, increased LP was noted following the irradiation of platelet concentrates (P < 0.05). These results suggest that the addition of various metal binding chelators may be effective in increasing the viability and longevity of stored platelet concentrates. PMID- 8239481 TI - Continuum analysis method applied to blood count reference intervals. AB - Conventional methods for extracting reference values from a sample of a purported normal population have a tendency to include some paranormal individuals and to obscure the existence of marginal or subclinical disorders. The suggestion is offered of defining the normal range of an analyte by extrapolation of the portion of its dispersion that fits a continuum. The method softens the present dichotomous character of the reference range. It narrows the normal range of some hematology parameters, suggesting that some paranormal individuals are excluded. PMID- 8239482 TI - Impairment of lymphocyte sensitivity to prostaglandin E2 in cultures from patients with lymphoadenopathy associated syndrome. AB - Prostaglandin E2 (PGE2) appears to have an immunosuppressive role in human immunodeficiency virus (HIV) infection. Therefore, the effect was studied of PGE2 pretreatment of T lymphocytes from patients with lymphadenopathy associated syndrome (LAS) on the expression of CD25 and CD71 as well as plaque forming cell (PFC) generation in pokeweed mitogen (PWM)-driven cultures. The PGE2-treated or untreated T lymphocytes were cultured with B cells and monocytes in the presence of PWM. Both CD25 and CD71 expression were assessed with an immunofluorescence technique; PFC generation was tested by hemolysis. Before exposure to PWM, LAS lymphocytes showed activation as evidenced by high CD25 and CD71 expression and PFC generation. Pretreatment by PGE2 did not inhibit expression of activation markers and PFC generation in LAS cultures, in contrast to what happened in control cultures. Thus, LAS lymphocytes are activated in vivo and are less sensitive to PGE2 inhibition than normal lymphocytes. PMID- 8239483 TI - Variation in gentamicin-induced death among independent cultures of proximal tubule cells. AB - This study determined the sensitivity of 12 independent isolates of cultured human proximal tubule (HPT) cells to gentamicin. The TD50 (dose [microgram per ml]) for each isolate was determined by linear regression analysis of cells exposed to a wide concentration range of gentamicin over a 16-day period of exposure. The results of this determination clearly showed that within a series of HPT cell cultures there was significant diversity in sensitivity to gentamicin. Three groups of toxic response were delineated. A single isolate, isolate I, was extremely resistant to gentamicin toxicity and was the sole member of group 1. The majority (9 of 12) of the isolates demonstrated intermediate sensitivity to gentamicin and were classified as group 2. Group 3 included two isolates that were very sensitive to gentamicin, with relatively low doses producing cell death. These results provide initial evidence that a portion of the variability seen in nephrotoxicity among patients receiving the aminoglycoside antibiotics might be mediated by factors intrinsic to each individual's proximal tubule cells. PMID- 8239484 TI - In vitro glycosylation in the retina in canine ceroid-lipofuscinosis. AB - Dolichol-dependent glycoprotein synthesis in the sensory retina of adult dogs with canine ceroid-lipofuscinosis (CCL) has been investigated by studying 3H mannose uptake in organ culture. Sensory retinal organ cultures were prepared from unaffected littermates and mongrel dogs were used as controls. Morphometric analysis of autoradiographs from dogs without disease showed maximal uptake predominantly into photoreceptors and inner segments, and no abnormalities of incorporation were observed in two affected animals. Tunicamycin blocked the reaction in both samples. Incorporation of 3H-leucine was also normal in CCL retinal tissues. These studies demonstrate that the pathways for addition of mannose residues into dolichol pyrophosphate prior to transfer onto acceptor proteins is similar in normal dogs and those with CCL and does not appear to be the primary lesion in ceroid formation. Its role as a model for lipopigment storage is discussed. PMID- 8239485 TI - Malassezia furfur folliculitis in cancer patients. The need for interaction of microbiologist, surgical pathologist, and clinician in facilitating identification by the clinical microbiology laboratory. AB - Malassezia furfur (MF) is a lipophilic yeast which can be found as a member of the indigenous microbiota of human skin. In immunocompromised transplant patients, MF can cause a distinctive folliculitis which is a clinical look-alike to Candida folliculitis, the latter of more potentially devastating significance. Recovery of MF in culture is dependent upon the addition to culture media of an exogenous source of fatty acids, such as olive oil. The addition of an extra Sabourauds plate with an olive oil overlay to the routine set of media used to inoculate all skin biopsy specimens in order to detect MF is labor-intensive and not cost-effective. Thus, MF may not be isolated in cases of MF folliculitis unless the clinical microbiology laboratory is put on alert by the clinical suspicions of the attending physician, or by histopathologic findings suggestive of folliculitis revealed by review of surgical pathology slides. The clinical, pathological, and microbiological findings of two cases of MF folliculitis are presented where an interactive approach featuring communication between the microbiologist, the surgical pathologist, and the clinician guided the microbiology laboratory to the isolation and identification of isolates of MF that were clinically-relevant. These cases underscore how a combined approach which features communication between the laboratory and the clinical services always provides superior guidance in the diagnosis and therapy of infectious diseases. PMID- 8239486 TI - Measurement of cocaine and metabolites in urine, meconium, and diapers by gas chromatography/mass spectrometry. AB - Analytical methods were evaluated for measuring cocaine (CO), benzoylecgonine (BE), and ecgonine methyl ester (EME) in urine and methanolic extracts from meconium and diapers by isotope dilution gas chromatography/mass spectrometry (GC/MS). Volatile derivatives of the extracted drugs were generated before GC/MS analysis. Methanolic extracts from meconium and diapers were reconstituted in drug-free urine and treated as above. The limit of detection for the GC/MS method was calculated to be approximately 11 ng per mL. Within-run coefficients of variation (CVs) for urinary CO, BE, and EME were 5.7, 5.3, and 11.4 percent, respectively (N = 10); corresponding CVs for meconium 6.4, 10.7, and 21.9 percent (N = 8). Quantitative results were linear from 25 to 10,000 ng per g of meconium and 25 to 5,000 ng per mL of urine. Day-to-day precision varied from eight percent (CV) for BE in refrigerated or frozen urine to 34 percent for EME in refrigerated meconium. Recoveries of CO, BE, and EME from urine were 63, 19, and 42 percent, respectively; corresponding recoveries from meconium were 64, 21, and 25 percent. Cocaine and metabolites were extracted from wet but meconium-free diapers into methanol, which was evaporated before reconstituting in drug-free urine and extraction on a solid phase column. The CO, BE, and EME were detected in previously drug-free meconium after portions were deposited in a diaper which was wet with drug-positive urine. Unless precautions are taken to prevent extracorporeal contamination of meconium with urine, concentrations of CO and metabolites in meconium may be substantially augmented by contamination with urine. Analysis by GC/MS of CO and metabolites extracted from diapers provides an attractive alternative to collection of urine, which is difficult and may cause discomfort for the neonate. PMID- 8239487 TI - [Lamprene in ano-perineal lesions of Crohn's disease. A retrospective study]. AB - The efficacy of clofazimine (Lamprene) was analysed retrospectively in twenty one patients with anoperineal lesions (APL) of Crohn's disease. Clofazimine is known for its antimycobacterial, antiinflammatory and immunomodifier properties. It is used with success in leprosy and certain dermatological disorders. A number of clinical and laboratory arguments suggest the probability of a role of mycobacteria in the etiology of Crohn's disease. METHODS: twenty one patients with ileo-colono-anal or bucco-colono-anal Crohn's disease formed the basis of this study. They had been treated in various ways for APL, without success. They had APL of varying degrees of advancement (primary lesions: seven cases; secondary lesions: ten cases; major advanced lesions: four cases). Treatment with Lamprene was given for three to 38 months (mean: 12 months), the mean cumulative dose being 40 grams (4 to 146). Other therapeutic measures were started simultaneously in twelve patients: medical in six cases, medico-surgical in four cases and surgical only in two cases. The aim of treatment in fourteen cases was to obtain the healing of ulcerated lesions and/or fistulas, in three patients to delay dilatation sessions and in four patients to avoid proctectomy in the short term. RESULTS: ten patients showed no improvement while eleven were improved (with regression of primary lesions in ten cases). Taking combined treatment into consideration, the link between the result obtained and Lamprene was considered probable in five cases, possible in four cases and uncertain in one case. Lamprene was well tolerated in general. It was not possible to evaluate the efficacy of treatment regarding intestinal disease. CONCLUSION: the efficacy of Lamprene in ano-perineal lesions of Crohn's disease is possible and is worthy of evaluation in a controlled trial. PMID- 8239488 TI - [Leiomyosarcoma of the small intestine presenting as an isolated lower digestive tract hemorrhage. Report of 2 cases]. AB - The authors report two cases of leiomyosarcoma of the small intestine revealed by lower and isolated digestive bleeding. In both cases, only the pelvic examinations by means of ultrasonography and/or CT scan have shown a mass which corresponded to the small intestine tumor. The authors underlined the necessity of US and/or CT scan pelvic when faced with a lower digestive bleeding whose etiology cannot be determined by the endoscopic or radiologic intestinal examinations. PMID- 8239489 TI - [Intestinal flora]. AB - The intestinal flora consists of a considerable mass of 10(14) cells which accompany the individual step by step from birth to death. It builds up multiple relations with the host housing it and represents a complex ecosystem, in dynamic equilibrium. The stability of the latter is maintained by virtue of only partially understood interactions. Gastric acidity, peristalsis and the immune system participate in this stability. However, this ecosystem is constantly subject to the actions of the outside medium, whether in the form of exogenous bacteria which are ingested on a constant basis of dietary substances. This fight against infection is one of the key factors in the role of the ecosystem. The actions of the flora are nevertheless far more extensive. It may also act on the host and, taking all populations into account, it is easy to see the enormous enzyme potential and the possibility of a range of metabolic effects, nutritional in particular. However, the state of equilibrium is fragile. Disturbances in the flora secondary to antibiotics and to bacterial infections cause disorders which are transitory as a rule. Equilibrium is most often restored by endogenous means (mucus, immune system, peristalsis). PMID- 8239490 TI - [Treatment of intestinal parasitoses]. PMID- 8239491 TI - [Parasitic fecal analyses. Prescription, application and interpretation of results]. AB - The increasing ease of intercontinental travel, immigration and the multiplication of cases of immune depression are such that European physicians are now frequently required to prescribe stool examinations for parasites. This article defines the laboratory approach to the diagnosis of intestinal parasites: indications for tests, based upon clinical and epidemiological data; methods of prescription and recommendations to patients; techniques used in the laboratory and their cost; minimum answer times; interpretation of results, in particular in the immune depressed. PMID- 8239492 TI - [Prognostic factors influencing healing of reflux esophagitis. A controlled trial of omeprazole versus ranitidine. Study group Omega]. AB - Four hundred and thirty patients with grade 2 or 3 esophagitis were treated after 2/1 randomization for 8 weeks with omeprazole 20 mg (n = 294) or ranitidine 150 mg bid (n = 136). Apart from treatment, 8 epidemiological factors (gender, age, occupation, obesity, smoking, alcohol, NSAID, and coffee or tea consumption), 5 clinical factors (day/night pain distribution, burning score, severity of regurgitation and of dysphagia, number of painful episodes requiring prescription of an antisecretory agent during the previous year, and onset of symptoms before age 30) and 3 endoscopic factors (grade and upward extension of esophagitis, and existence of hiatal hernia > or = 5 cm) were analysed. The influence of these factors on healing at 8 weeks and on changes in symptoms was evaluated by multivariate analysis. 92.1% of patients enrolled were analyzed. In comparison with ranitidine, omeprazole increased the percentage of healed patients (93% v. 67.5%, p < 0.001) and the rapidity of disappearance of symptoms (5 days v. 7 days, p < 0.001). Independent good prognostic factors associated with healing rate were treatment with omeprazole (p < 0.001) and grade 2 esophagitis (p < 0.001) while those associated with the disappearance of symptoms were a low burning score (p = 0.001), advanced age (p = 0.004), treatment with omeprazole (p = 0.005), the absence of any occupation (p = 0.01) and male gender (p = 0.017). The results of this study show that, apart from treatment, endoscopic factors are predictive of the healing of reflux esophagitis treated by antisecretory agents while clinical factors are more important with regard to the disappearance of symptoms. PMID- 8239493 TI - Examination of DNA methylation of chromosomal hot spots associated with breast cancer. AB - Widespread hypomethylation of DNA and regional hypermethylation, including tumor suppressor regions, have been demonstrated in several human cancers. Since a highly heterogeneous array of genetic anomalies have been associated with breast cancer, we examined several chromosomal hot spots for abnormal methylation patterns. Low-levels of increased methylation of HRAS (11p15) were observed between normal and tumor breast tissue samples from 8 patients. No noticeable variation in methylation was observed with DNA probes from chromosomes 11p15, 1p36, 17q22, 17p13.3 and 3p21 for the 7 ductal breast carcinoma patients, though some variability was observed for a patient with atypical medullary carcinoma. Additionally, the methylation pattern of the estrogen-receptor gene (6q24-27), whose protein product is increased in numerous breast cancers, also did not change. Therefore, as opposed to other cancer types, widespread hypomethylation and regional hypermethylation do not appear to be involved in the early stages of breast cancer and does not account for the molecular heterogeneity of the disease. Proposed alternative mechanisms for the diversity of genetic alterations associated with breast cancer are discussed. PMID- 8239494 TI - Activation of a calcium magnesium independent endonuclease in human leukemic cell apoptosis. AB - Cytotoxic drugs induce apoptosis in human tumour cell lines and this is characterised by fragmentation of the cell's DNA into nucleosome size units or multiples thereof. In the present study we demonstrated that nuclei isolated from three human haematopoietic cell lines, HL-60, U937 and K562, contain an endonuclease that is independent of Ca++, Mg++ and Na+ ions for its activity. This contrasts with what has previously been shown for a number of rodent cell types in which apoptosis has been studied. The lack of ion sensitivity is also found in the nuclei of peripheral blood granulocytes, indicating the data are not peculiar to cell lines. In addition, this particular endonuclease activity does not appear to be sensitive to the endonuclease inhibitor aurintricarboxylic acid. The previously demonstrated lack of calcium flux in HL-60 cells undergoing apoptosis, and the current demonstration of a lack of an endonuclease dependent on this ion for its activity, suggest that the mechanism of apoptosis in human cells may be different from that in rodent cells. PMID- 8239495 TI - Growth of prostatic cancer cells, DU 145, as multicellular spheroids and effects of estramustine. AB - The human prostatic carcinoma cell line DU 145 was grown as multicellular spheroids in vitro. The volume doubling time during the early exponential growth phase was about 5 days. The saturation volume, in the plateau phase of the growth curve, was in the order of 1.4 mm3. The spheroids developed a central degenerative region surrounded by a 0.1-0.3 mm layer of viable cells. The DU 145 spheroid system is planned to be used as a model in studies on chemotherapy and targeted radiotherapy of micrometastases of prostatic cancer. Some effects of the drug estramustine, EM, a conjugate of estradiol and nornitrogen mustard, were analysed in this introductory study. Tritium-labelled estramustine, 3H-EM, bound both in the viable cell layers and in the degenerative region of the spheroid already after 1 hour of incubation which indicated good penetration. The viable cells bound only low levels of 3H-EM while the degenerative region bound 3H-EM to a higher extent. The amount of bound 3H-EM increased after incubation for 24 hours. The binding was nonspecific since it could not be inhibited by pretreatment with an excess of non-radioactive EM. Furthermore, 3H-EM bound to a similar extent in glioma and colon carcinoma spheroids used for comparison. Incubation of DU 145 spheroids for 24 hours with EM (20 mg/ml) induced a growth delay of 6-7 days and a transient increase in the volume of the extracellular spaces for a few days following the treatment. The results showed that the binding of EM to prostate DU 145 cells growing as spheroids was not specific and that the toxic action was limited. An interesting result was that EM works as an extracellular space expander. This might be exploited in combination treatments with other agents. PMID- 8239496 TI - Collateral sensitivity to thaliblastine and/or hyperthermia exhibited by a rat ovarian tumor cell line selected for resistance to cisplatin. AB - Drug resistance severely limits the effectiveness of clinical cancer chemotherapy. Employment of drugs other than the selected compounds with different mechanisms of action may provide a potential way to improve the therapeutic effects. Thaliblastine (TBL), a natural compound, showed a 2-fold higher cytotoxicity in a cisplatin (DDP) resistant rat ovarian tumor cell line (0 342/DDP) than in its parental sensitive line (0-342), as determined by an antiproliferation assay with 24 h continuous exposure. This phenomenon was also observed following 2 h pulse exposure if combined with heat treatment (40 degrees C). Further escalation of the temperature to 43 degrees C alone brought about 74.7 +/- 17.0% growth inhibition in the sensitive and 97.2 +/- 1.8% in the resistant line. Under this condition, the ID50 of TBL was again only half as much in 0-342/DDP cells as in the parental cells (12 vs 24 micrograms/ml) when compared to the hyperthermic treatment alone. In a colony formation assay with 2 h pulse exposure, the hypersensitivity of the resistant cells to DDP and/or heat was further confirmed. Alkaline elution showed that 24 h continuous treatment with TBL induced DNA single-strand breaks (SSB) in a dose-dependent manner in 0 342/DDP cells, whereas there was almost no DNA-SSB production by TBL in the sensitive line, possibly in part accounting for the hypersensitivity of the DDP resistant cells to TBL. The heat treatment (40 degrees C for 2 h) induced SSB in both lines, which was further enhanced by combination with TBL. This damage was repaired in part in 0-342 but almost completely in 0-342/DDP line after cells grew in drug-free medium for 48 h following the exposure, indicating that resistant cells can more efficiently repair DNA damage by either TBL or hyperthermia. Altogether, these results suggest that TBL may have potential to be used clinically as an alternative in the treatment of cisplatin-resistant malignancies with hyperthermia. PMID- 8239497 TI - Different binding properties of three monoclonal antibodies to sialyl Le(x) glycolipids in a gastric cancer cell line and normal stomach tissue. AB - We established a mouse monoclonal antibody (Mab) KM93 which recognized sialyl Le(x)-carbohydrate epitope determined by solid phase radioimmunoassay using a panel of authentic glycolipids. The specificity of KM93 was similar to another anti-sialyl Le(x) Mab CSLEX-1 established previously, and different from that of Mab FH6 which recognized sialyl Le(x)-i (sialyl dimeric Le(x)). In a further study, however, we found that KM93 reacted with some glycolipids much more strongly than CSLEX-1 did on thin-layer chromatography (TLC) plates. We purified two gangliosides named K-1 and K-2 from gastric cancer cell line KATOIII, and three gangliosides named H-1, H-2, and H-3 from human stomach. KM93 reacted with all of these glycolipids. CSLEX-1 reacted with K-1 and K-2 with less intensity than KM93 did, and faintly reacted with H-1, but not at all with H-2 or H-3. FH6 did not react with K-1, K-2 or H-1, while it stained H-2 and H-3. In spite of this different reactivity with Mabs, analysis by proton nuclear magnetic resonance (1H NMR) proved that carbohydrate structure of K-1 and H-2 were the same: NeuAc alpha 2-->3Ga1 beta 1-->4 [Fuc alpha 1-->3] G1cNAc beta 1-->3 Ga1 beta 1-->4G1c beta 1-->1Cer. The H1 NMR spectrum of H-3 also indicated that H-3 consisted of the same sugars as K-1. These results indicated that KM93 had wider reactivity than CSLEX-1, and that the distinct reactivity of KM93 from CSLEX-1 was not caused by sugar moiety. It was also shown that the interaction of sialyl Le(x) sugar determinant with MAb depended on the tissue origin of molecules carrying carbohydrate. PMID- 8239498 TI - Effects of nicotinamide on clonal rat insulinoma cell proliferation, polyamine content and insulin secretion. AB - Tumoral insulin-secreting cells (RINm5F) were cultured for 4 days with nicotinamide, a poly (ADP-ribose) synthetase inhibitor. This resulted in a dose dependent inhibition of the cellular proliferation rate, along with a modest decrease in cell viability. Nicotinamide failed to affect the cellular insulin content, but reduced the insulin secretion into the culture medium. Nicotinamide treatment furthermore reduced the cellular content of polyamines, compounds seemingly involved in regulation of the growth and function of insulin-producing cells. Restoration of the decreased polyamine content by the concomitant addition of the polyamine precursor putrescine failed to influence the decreased proliferation rate and insulin secretion, indicating that the inhibitory effects of nicotinamide on these functions cannot be ascribed to the decrease in polyamine content. PMID- 8239499 TI - Intracavitary treatment of malignant ascitic carcinomatosis with oily anticancer agents in rats. AB - The anticancer effects and pharmacokinetics of intraperitoneal (ip) injection were determined for doxorubicin (DOX) dissolved in a lymphographic oily contrast medium, Lipiodol Ultrafluid (Lipiodol) (DOX/Lipiodol), using AH 130 ascitic tumor in rats. A high percentage (50%) of long-term survivors (cure) was observed with only a single ip injection of the DOX/Lipiodol on the seventh day after tumor inoculation, by which time the tumor ascites accumulated to a level similar to that seen clinically. The cure rate of this lipid formulation was much better than that for DOX dissolved in saline (DOX/saline) (17%). An immunopotentiating agent, Picibanil, given in 1 KE (clinical unit) or 5 KE doses was no better than the saline group, indicating that the above effect was not a result of immunopotentiation by Lipiodol. The severity of toxic side effects of DOX was also reduced by lipid solubilization. Pharmacokinetic study showed that DOX/saline was absorbed rapidly into the blood stream from the peritoneal cavity, whereas DOX/Lipiodol was retained in the ascites at a higher concentration for a much longer time. Thus the present results suggest that lipid formulations of anticancer agents have augmented therapeutic efficacy by intracavitary injection. The lipid formulations show increased stability in and prolonged slow release from the lipid milieu. Thus this therapeutic tactic of using oily anticancer agents appears to be promising for the control of pleural and peritoneal carcinomatoses. PMID- 8239500 TI - The measurement of glucose consumption in histoculture to determine effects of doxorubicin and cisplatinum on human gastric carcinoma. AB - We have developed a chemosensitivity assay of human tumors growing on collagen sponge-gel-supported histoculture. This assay is thus termed the Histoculture Drug Response Assay (HDRA). In the HDRA, the end points of [3H]thymidine incorporation measured by histological autoradiography and tetrazolium dye reduction were initially used and found to have good in vitro-in vivo correlations, including that determined in clinical trials. We have now developed glucose consumption as an endpoint in histoculture. We have monitored glucose consumption for 11 weeks with histocultured stomach cancer tissue that was obtained from a patient with stomach cancer metastatic to the lymph node. The histocultured lymph node specimens were treated with various concentrations of doxorubicin and cisplatinum. The glucose-consumption rate decreased with greater concentrations of both drugs. The results correlated with the thymidine labeling index. From these results, we conclude that the glucose-consumption-rate endpoint in histocultured cancer tissue is non-destructive, unlike the [3H]thymidine and tetrazolium dye end points, allowing serial determinations over extended periods in culture. Thus, the glucose consumption end point may enhance the development of optimal treatment doses and schedules. We also conclude that long-term histoculture drug response studies of metastatic stomach cancer are possible. PMID- 8239501 TI - The analysis of ifosfamide and its metabolites (review). AB - Ifosfamide is a member of the oxazaphosphorine class of cytostatic drugs. Ifosfamide has to be metabolized prior to expressing its cytotoxicity. This metabolism leads to a great variety of metabolites. An overview is given of the methods of analysis of ifosfamide and its metabolites as well as the current knowledge about its metabolism, toxicity and clinical use in relation to the bio analysis. For ifosfamide and some of its metabolites reliable methods of analysis exist. However, for the most important metabolites, isofosforamide mustard and acrolein, progress has to be made in improving the simplicity and sensitivity of the existing methods. PMID- 8239502 TI - Differential effects of 12-O-tetradecanoyl-13-phorbol acetate (TPA) on growth, migration and invasion of a human glioma cell line. AB - To evaluate the effects of protein kinase C (PKC) activation on a human glioma cell line (GaMG), this study examined the effects of the phorbol ester 12-O tetradecanoyl-13-phorbol acetate (TPA) on the tumor properties proliferation, migration and invasion. GaMG monolayer growth was reduced by 30% (p < 0.05) in the presence of TPA and, concomitantly, cells accumulated in the G2M phase of the cell cycle as determined by flow-cytometric analyses. In studies of directional migration of cells from GaMG spheroids, low concentrations of TPA increased the area of migration by 60% (p < 0.005). The effects at higher concentrations of the phorbol ester were still present but less pronounced. Glioma invasion was assessed with a glioma spheroid/brain aggregate co-culture technique and revealed TPA-induced enhancement of the invasive process (p < 0.005) at low concentrations of the phorbol ester. Down-regulation of PKC by passaging the cells in a TPA containing medium significantly decreased the migratory response to TPA. Moreover, the PKC inactive phorbol ester 4-alpha-phorbol-12,13-didecanoate had no effect on the migratory response of GaMG cells. It is concluded that metabolic pathways under the control of PKC may play an important role in the control of glioma growth, migration and invasion. The effects of PKC activation are probably dependent on the growth conditions. Studies on PKC activation under different culture conditions in three-dimensional systems are required to evaluate further the role of PKC in glioma biology. PMID- 8239503 TI - Protein kinase C is not involved in cholesterol-induced resistance to synthetic ether lipids. AB - The possible role of protein kinase C in cholesterol-induced resistance to ether lipids was investigated. The enrichment of HL60 cells in cholesterol (CHOL) (HL60 CHOL) resulted in a significant increase in the ID50 values for 1-octadecyl-2 methyl-rac-glycero- 3-phosphocholine (ET-18-OMe) (3.75 +/- 0.7 microM and 6.69 +/ 0.5 microM for HL60 and HL60-CHOL, respectively). In the same conditions, HL60 and HL60-CHOL cells showed comparable levels of both cytosolic and membrane associated protein kinase C activity. Phorbol ester (PMA) stimulation induced protein kinase C to translocate from the cytosol to the plasma membrane in both cell types and with similar kinetics (272 +/- 32% and 299 +/- 41% increase in HL60 and HL60-CHOL, respectively after 100 ng/ml PMA for 10 min). Pretreatment of the two cell types with 50 microM ET-18-OMe resulted in comparable levels of PKC inhibition after phorbol ester stimulation. These results suggested that alterations in plasma membrane lipid composition induced by CHOL do not result in major changes in protein kinase C activity. Thus, protein kinase C does not appear to be involved in cholesterol-induced resistant phenotype in HL60 cells. PMID- 8239504 TI - Combined modulation by leucovorin and alpha-2a interferon of fluoropyrimidine mediated growth inhibition. AB - One way to improve fluoropyrimidine activity is the use of leucovorin (LV). Another way is the use of alpha-2a interferon (alpha-IF). The mechanism of the alpha-IF effect on fluoropyrimidines has not yet been elucidated. Besides, only limited data area available on double modulation (LV and alpha-IF) of fluoropyrimidines. Therefore, the modulating capacity of both drugs was tested in a fluoropyrimidine resistant (COLO 320) and a sensitive (SW 948) cell line. Also, the binding capacity of thymidylate synthase (TS) to 5-fluoro-2'-deoxyuridine-5' monophosphate (FdUMP) and TS catalytic activity were studied in both cell lines as well as the effects of 5-fluorouracil (5-FU) and alpha IF on enzyme activity. COLO 320 had, compared to SW 948, a 7.5 fold higher FdUMP binding capacity to TS. TS activity was 4.4 and 11.3 fold higher at 10 and 1 mM substrate, respectively. In COLO 320 enhancement of 5-FU, either by LV or by alpha-IF, was not possible. Since LV did enhance 5-fluoro-2' deoxyuridine (FUdR) activity, it is conceivable that 5-FU mediated growth inhibition in COLO 320 is not TS mediated. SW 948 was sensitive to both modulating agents with a 2.4 fold lower IC50 for 5-FU/LV, 6.8 fold lower IC50 for 5-FU/alpha-IF and a 11.2 fold lower IC50 for 5-FU/LV/alpha IF. Effects of LV and alpha-IF on FUdR were comparable but less pronounced, with a 3.4 fold lower IC50 for FUdR/LV/alpha-IF compared with FUdR alone. Thymidine, which circumvents TS inhibition, neutralized the synergistic effects of alpha-IF, indicating that alpha-IF enhancement is mediated via inhibition of DNA synthesis. However, no direct effects of alpha-IF on FdUMP binding or catalytic activity could be demonstrated. IN CONCLUSION: alpha-IF can increase 5-FU/LV mediated growth inhibition in fluoropyrimidine sensitive colorectal cancer cells. FdUMP binding capacity and catalytic activity of TS may predict sensitivity to (modulation of) fluoropyrimidines. PMID- 8239505 TI - Genomic abnormalities in hepatocarcinogenesis. Implications for a chemopreventive strategy. AB - Carcinogenesis is a complex process characterized by the cumulative activation of various oncogenes and the inactivation of suppressor genes. Epigenetic mechanisms are also involved. Mutational activation of ras family genes occurs in most spontaneous or carcinogen-induced liver tumors, in susceptible mice, and less frequently in preneoplastic lesions. This suggests a pathogenetic role of these changes in hepatic carcinogenesis, in the mouse. Overexpression of various growth related genes occurs in preneoplastic tissue during rat liver carcinogenesis, but mutational activation of protooncogenes, notably of ras family genes, seems to be a late and rare event, while c-myc amplification is a late but frequent event in both rodent and human carcinogenesis. However, mutation of the suppressor p53 gene has been found in relatively early preneoplastic lesions in rat liver, and it may be frequently seen in human hepatocellular carcinomas. The possibility that this mutation is involved in the initiation stage of liver carcinogenesis is an attractive hypothesis which needs further evaluation. DNA hypomethylation is involved in carcinogenesis, but the mechanisms underlying this effect are still elusive. Hypomethylation of growth-related genes is associated with their overexpression and this could favor overgrowth of preneoplastic liver tissue. Decrease in S-adenosyl methionine/S-adenosylhomocysteine (SAM/SAH) ratio occurs in the liver of rats fed a methyl deficient diet, which is a carcinogenic treatment, and in preneoplastic liver tissue, developing in initiated/promoted rats fed an adequate diet. The role of low SAM/SAH ratio in carcinogenesis is substantiated by the tumor chemopreventive effect of lipotropic compounds. Treatment with exogenous SAM prevents the development of preneoplastic and neoplastic lesions in rat liver. This is associated with recovery of SAM/SAH ratio, DNA methylation and inhibition of growth-related gene expression. SAM effect on prenoplastic cell growth is abolished by 5-azacytidine, a hypomethylating agent, indicating the involvement of DNA methylation. The possibility that in SAM-treated rats, methylation and inhibition of the expression of growth-related genes is implicated in growth restraint is attractive and should be further evaluated. Modulation of rat liver carcinogenesis by influencing gene expression through DNA methylation or other epigenetic mechanisms could be a new approach to chemoprevention of these tumors. PMID- 8239506 TI - The statistical analysis of cancer inhibition/promotion experiments. AB - The purpose of this paper is to address the very important problem of accurate statistical analysis of certain types of cancer inhibition/promotion (IP) experiments. These experiments are routinely used by the National Cancer Institute to test the effects of potential chemopreventative agents. The statistical analysis is difficult since there is Type I censoring. In the IP experiments under investigation, laboratory animals (rats) are injected with a single dose of either a direct or indirect acting carcinogen. In the mammary tumor system, animals in the control group generally develop 5-7 tumors and typical experiments are usually terminated after 4-6 months. Animals are sacrificed at the end of the experiment and all observed tumors are confirmed. The two most common response variables are the number of observed tumors per animal and the rate of tumor development. The difficulty in analyzing these experiments occurs because experiments are terminated before all induced tumors have been observed. Fewer observed tumors in one group compared to another could be the result of a decreased number of induced tumors, a decrease in growth rate, or a combination of both. It is essential for the experimenter to distinguish between these two different biological actions. Present statistical techniques do not account for this confounding and since they rely primarily on nonparametric procedures, do not present an accurate description of potential IP agents. In this paper we introduce a parametric procedure that explicitly acknowledges the confounding present in experiments of this nature. The analysis is based on the comparison of the mean number of tumors per group (lambda) and the mean time to tumor appearance (mu). A longer mean time to development is believed to indicate a slower tumor growth rate. Hypothesis tests are developed to determine if there is an overall experiment effect, to isolate which groups are contributing to an observed experiment effect, and to isolate factors (tumor number and/or growth rate) contributing to an observed group difference. Confidence regions for (lambda, mu) are also generated. This analysis leads to a better understanding of how potential IP agents function. PMID- 8239507 TI - Overexpression of c-erbB-2 and c-myc but not c-ras, in canine melanoma cell lines, is associated with metastatic potential in nude mice. AB - Overexpression of the proto-oncogenes c-erbB-2, c-myc, and c-ras have been associated with neoplastic transformation in a variety of tumours. We investigated expression of these oncogenes in 5 canine melanoma cell lines and 6 clonal derivatives of 1 of the cell lines, CML-6M, to determine what impact overexpression had on tumour cell growth and metastatic potential. All 11 cell lines were tumourigenic at subcutaneous inoculation sites in nude mice, but spontaneous metastasis to lung was a characteristic of only the CML-6M cell line and 3 of 6 clonal derivatives of CML-6M. Investigation of oncogene overexpression revealed no obvious pattern of expression among the 5 tumour-derived cell lines whereas overexpression of c-erbB-2 and c-myc was consistently found in the 3 clonal cell lines characterized by high metastatic potential, and in primary and metastatic mouse xenografts induced by these lines. This data suggests involvement of overexpression of these genes in development of canine melanoma and associates their overexpression with metastatic potential in nude mice. PMID- 8239508 TI - The myc family of oncogenes and their presence and importance in small-cell lung carcinoma and other tumour types. AB - The myc family of cellular oncogenes, c-myc, N-myc, encodes three highly related, cell cycle specific, nuclear phospho-proteins. All are able to transform primary rat embryo fibroblasts when cotransfected with the c-ras oncogene. Myc family genes are differentially expressed with respect to tissue type and developmental stage and dramatic changes in their expression coincide with critical developmental transitions in many cell lineages. Their conservation as distinct genes over a large phylogenetic distance and their unique pattern of expression in developing embryos support the view that each member of the myc family may have an important and discrete biological activity. The nuclear localization and the presence of contiguous regions containing the basic region/helix-loop helix/leucine zipper motifs suggest that myc-encoded proteins are involved in gene expression during growth and differentiation. The role of myc-oncogene amplification and/or overexpression in tumourigenesis is still not clear, but application of new biotechnological tools as transfection vectors and the antisense strategy may lead to a better understanding of the importance of the myc family of oncogenes. Transfection of cells with oncogenes in expression vectors conferring high levels of oncoprotein are used to analyze the influence of overexpression of the oncogene, while incubation with an appropriate antisense oligonucleotide leading to an inhibition of oncogene expression may give some clues to the necessity, function and importance of the oncogene. These two techniques have proven to be useful tools in the illumination of the function of these ubiquitous (proto-) oncogenes and probably will help to clarify at least a part of their role in, for instance, apoptosis, tumourigenesis and the development of resistance to antitumour drugs. PMID- 8239509 TI - Retinoid modulation of c-myc and max gene expression in human breast carcinoma. AB - Retinoic acid (RA) modulation of c-myc and max gene expression was investigated in human breast carcinoma (HBC) cell lines. Our results demonstrate that c-myc and max genes are differentially expressed in HBC cells which was accompanied by increases in [3H]-thymidine incorporation and percent of cells in the S phase of cell cycle. RA-mediated increase in c-myc mRNA levels were noticed as early as 30 min. and maximum levels were attained by 1 h. RA effect on max mRNA levels was slow and gradual with the maximum effect noticed by 48 h. Nuclear run-on assays demonstrated that RA mediated its effect by increasing the rate of transcription of both of these genes. We thus report for the first time that RA, during its growth inhibitory effects on MCF-7 HBC cells, positively regulates the gene expression of c-myc and max. PMID- 8239510 TI - Selective cytotoxicity of L-glutamic acid gamma-monohydroxamate (GAH) for melanoma tumor cells. AB - We have previously shown that L-glutamic acid gamma-monohydroxamate (GAH) exhibits an antitumor activity, both in vitro and in vivo. In this report we explore the selective cytotoxicity of GAH in vitro by comparing the survival of tumor and normal cells. GAH exerts an irreversible delayed effect with tumoral cells and a reversible effect with normal cells: after a short incubation time of 6 hrs in the presence of 1.2 mM GAH and after removal of the drug, the survival of N Ter Dau and MRC5 cells was identical reaching about 85% after 24 hrs of culture. But, after another 48 hrs of culture, MRC5 cells recovered 100% cell survival while with N Ter Dau cells the survival decreased to 65%. A longer exposure time to GAH (18 hrs) and an additional 54 hrs of culture after removal of GAH led to 50 +/- 10% of cell survival with normal cells but only 25 +/- 10% with tumor cells. Using a long-term clonogenic assay, we showed that the 25% N Ter Dau cells surviving at 72 hrs after GAH treatment led mainly to abortive colonies (17% +/- 3%) with only 2.3 +/- 0.9% of surviving colonies. Such a difference does not exist for normal cells. Cell cycle analysis of tumor and normal cells treated with GAH (18 hrs, 1,2 mM) has shown that the drug prevents both cell type from cycling from G1 to S phase. However, the two cell types started to cycle again after removal of GAH but a delay of 24 hrs was observed for tumoral cells compared to normal cells. PMID- 8239511 TI - Growth factor regulation of 17 beta-hydroxysteroid dehydrogenase activity in a human ovarian cell line: modulation by 17 beta-estradiol. AB - We have examined the effects of IL-6 and bFGF on growth and reductive 17 beta hydroxysteroid dehydrogenase (17-HSD) activity in a human ovarian cell line A2780 under defined culture conditions (standard RPMI (GM+PHR), phenol red-free RPMI (GM-PHR), or GM-PHR supplemented with 17 beta-estradiol). In media GM+PHR, neither factor influenced terminal cell density. However there was a 2-fold stimulation in 17-HSD activity (E1- > E2) in the presence of IL-6 and a 3-fold increase in the presence of bFGF. Cells cultured GM-PHR showed a 25-30% reduction in terminal cell density compared with cells cultured GM+PHR, and addition of factors resulted in a modest reduction (10-15%) in cell growth, with no detectable changes in 17-HSD. Addition of 17 beta-estradiol to GM-PHR cultures increased terminal cell density by up to 3-fold, with no effect on 17-HSD, and when combined with IL-6 or bFGF, a synergistic effect on growth was observed, but 17-HSD levels were unchanged. Our results suggest that cultured ovarian tumour cells respond to paracrine factors likely to be present in the ovary in vitro, and there is a possible interactive effect of estromedins with these factors. PMID- 8239512 TI - State of p53, Rb and DCC tumor suppressor genes in human oral cancer cell lines. AB - The tumor suppressor genes p53, Rb, and DCC were studied in five human oral cancer cell lines (FaDu, SCC-4, HEp-2, 1483, and OEC-M1) and in primary normal human oral keratinocytes (NHOK). All tested cancer lines had similar amount of p53 messages to normal cells, but the cancer lines FaDu and SCC-4 contained significantly higher p53 protein levels than did the normal counterpart. Sequencing p53 cDNA for these cancer cells showed point mutations: In the FaDu cell line, a mutation of CGG to CTG occurred at codon 248; and in the SCC-4 cell line, a mutation of CCC to TCC occurred at codon 151. The HEp-2 and 1483 cancer lines translated very low levels of p53 protein compared to the normal counterpart. Sequencing of p53 cDNA for HEp-2 and 1483 lines showed no mutations. Southern and Northern analyses revealed that these cell lines harbored HPV-18 DNA and expressed the viral E6/E7 protein. The OEC-M1 line showed different restriction fragment length polymorphism for the p53 gene compared with other cells, and did not express p53. All oral cancer cell lines except the OEC-M1 cells expressed both phosphorylated and hypophosphorylated Rb proteins. Further, the OEC-M1 line expressed smaller sized hypophosphorylated Rb proteins compared with normal cells. Unlike the other cancer lines, the HEp-2 and OEC-M1 lines also did not contain DCC mRNAs. These data indicate that "high risk" HPV infections and mutations of p53, Rb, and DCC genes are frequently found in oral cancer cells and may be associated with oral cancer. PMID- 8239513 TI - Cytogenetical observations in nine ocular malignant melanomas. AB - The cytogenetical findings in one conjunctival and eight uveal melanomas are reported. Six of the intraocular tumors displayed clonal abnormalities. In two of these cases there were both extensive numerical and structural aberrations. Only the single ciliary body melanoma showed the deviations monosomy 3 and gain of 8q considered typical of uveal melanomas. No preferential pattern could be discerned for the clonal abnormalities in the other uveal melanomas. The one conjunctival tumor, arising in an area of precancerous melanosis, showed only a small clone with a 4q+marker. The clonal abnormalities in the nine cases showed no similarities to those on record for cutaneous melanomas. PMID- 8239514 TI - Inhibition of tumor cell invasion in the Boyden chamber assay by a mannosidase inhibitor, mannostatin A. AB - An alpha-mannosidase inhibitor, mannostatin A, from Streptoverticillium verticillus var. quintum inhibited chemotactic invasion of mouse B16/F10 melanoma cells in the Boyden chamber assay. It also inhibited in vitro invasion of K-ras NIH3T3 cells. Mannostatin A did not inhibit the growth of either cell line at the concentration effective to inhibit invasion. Addition of mannostatin A to the cultured B16/F10 or K-ras-NIH3T3 cells inhibited cellular alpha-mannoside activity specifically. Mannostatin A-treated B16/F10 cells also showed decreased metastatic activity in vivo in C57Bl/6 mice. PMID- 8239515 TI - In vivo use of N-(4-hydroxyphenyl retinamide)-0-glucuronide as a breast cancer chemopreventive agent. AB - The inhibitory effects of N-(4-hydroxyphenyl) retinamide and its glucuronide derivative on the development and growth of 7,12-dimethylbenz (a) anthracene - induced rat mammary tumors in vivo were compared. The results indicate that the glucuronide had a greater chemopreventive potency than equimolar concentration of the free retinoid by all the criteria measured, mainly the inhibition of tumor incidence, multiplicity and tumor growth. HPLC analysis of the blood of the rats showed no hydrolysis of the glucuronide during its chronic consumption, indicating that the retinoid glucuronide is probably acting in vivo per se rather than through hydrolysis to the free retinoid. The higher potency and lower toxicity of the glucuronide suggests, for the first time, that the conjugate may have an in vivo chemopreventive advantage over the parent retinamide. PMID- 8239516 TI - Enhancement of mitomycin-C cytotoxicity by combination with flavone acetic acid in a murine tumour. AB - The antitumour activity of a combination of Mitomycin C (MMC), a prototypical bioreductive agent, with the vascular occluding agent, flavone acetic acid (FAA), was examined in a murine tumour. Flavone acetic acid induced a long lived reduction of blood flow in the tumour thereby inducing hypoxia and cell death by ischaemia. Dose response curves for tumour cell survival and regrowth delay were constructed after each agent administered alone or when combined. The greatest antitumour effect was measured when the two agents were administered simultaneously. The reduced tumour perfusion induced by the combination was not lower than that for FAA alone indicating that the greater in vivo effect was not due to enhancement of the vascular occluding effect of the FAA. The role of the tumour microenvironment in determining the MMC cytotoxicity was examined using in vitro incubations of MMC under controlled oxygenation and pH. MMC cytotoxicity was increased in hypoxia (enhancement ratio = 2.0) and also when extracellular pH was reduced from 7.4 to 6.4 (enhancement ratio = 1.5). Since the microenvironmental consequences of induced ischaemia are reductions in both oxygenation and pH these parameters are probably responsible in large part for the enhancement of cytotoxicity observed when MMC and FAA are combined. PMID- 8239517 TI - A possible direct action of a gonadotropin-releasing hormone agonist, goserelin, on the clonogenic growth of human breast cancer. AB - Direct inhibitory effects of a gonadotropin-releasing hormone agonist (GhRh agonist, goserelin, GOS) were studied on the in vitro clonogenic growth of human breast cancer, in comparison with estradiol-17 beta (E2) and tamoxifen (TAM). Estrogen (ER) and progesterone receptors (PgR) were asseyed in the adjacent cancer tissues using the DDC method. In 19% or 12 out of 64 cancers, E2, 10(-8) M, was shown to be effective in increasing the number of colonies per dish to 150% or more of the control. On the other hand, 16% (10/64) responded to TAM or GOS, respectively, i.e. the plating efficiency decreased to 50% or less of the control. While E2 and TAM had a tendency to have better effects in ER-positive tumors than ER-negative ones, the inhibitory effect of GOS was not dependent on the presence or absence of ER or PgR in the tumor. The GOS sensitivity did not correlate with the E2 dependence or the TAM sensitivity. These results suggest that a possible direct action of GOS may not be mediated by the ER system. PMID- 8239518 TI - Effect of an aryl chloroethyl urea on tubulin and vimentin syntheses in a human breast cancer cell line. AB - A new class of antineoplastic agents, 1-aryl-3-(2-chloroethyl) ureas (CEUs), was recently developed in our laboratory. To optimize the pharmacological and the biological properties of this new class of compounds and to determine its mechanism of action, at the cellular level, we studied the effect of 4-tert-butyl [3-(2-chloroethyl) ureido] benzene (tBCEU) on MDA-MB-231, a human breast cancer hormone-independent cell line. The effect of tBCEU on protein synthesis and on the accumulation of specific mRNAs was evaluated. The results indicate that tBCEU increases the synthesis of at least two proteins present in the cytoskeleton: tubulin and vimentin. The effect of tBCEU on their transcripts indicates that tBCEU decreases the accumulation of tubulin and vimentin mRNA. These results suggest that the antineoplastic activity of tBCEU is in part related to an alteration in the synthesis pathway of tubulin and vimentin. PMID- 8239519 TI - A somatostatin analogue (SMS 201-995) alters the toxicity of 5-fluorouracil in Swiss mice. AB - The somatostatin analogue 201-995 (SMS), given in a short treatment schedule, 1 microgram/mouse, days 1-4, increased the lethal toxicity of a single, day 2, low lethal dose of 5-fluorouracil (FU), 400-500 mg/kg, in Swiss mice. The analogue did not change the time course of toxic deaths and produced neither autopsy evidence of increased gastrointestinal nor of increased bone marrow toxicity. Paradoxically, the analogue protected the peripheral white blood cell count (WBC). The nadir was only half as severe in the animals treated with SMS in addition to FU. The mechanism responsible for more frequent deaths due to combining SMS with FU is unknown. That brief administration of SMS in combination with marginally lethal, intensive therapy can produce unexpected increase in animal deaths bears consideration because these drugs are sometimes given simultaneously to patients, either fortuitously or in trials designed to limit the gastrointestinal side effects of FU. PMID- 8239520 TI - Gene therapy of cancer: use of IL-2 gene transfer and kinetics of local T and NK cell subsets. AB - Experiments were designed to compare the efficacy of recombinant IL-2 immunotherapy and IL-2 gene therapy of i.p. growing murine plasmacytoma X63 Ag8.653. The kinetics of peritoneal exudate mononuclear cells were monitored during the progression and gene therapy of the plasmacytoma, using cytofluorometric analysis and monoclonal antibodies against T and NK cell subsets. It has been found that the percentage of mice protected against plasmacytoma transplants was higher in mice treated by transfer of genetically manipulated IL-2-producing plasmacytoma cells as compared to the mice repeatedly injected with recombinant IL-2. Intraperitoneal inoculation of the X63-Ag8.653 plasmacytoma led in most of the inoculated mice to an increased percentage of NK+, ASGM1+, Thy 1.2+, CD3+ and TCR alpha beta+ cells in the peritoneal fluid. The presence of macroscopically detectable i.p. tumours was accompanied by a higher increase in the percentage of NK+ and TCR gamma delta+ cells. Local IL-2 gene therapy of the plasmacytoma either prevented or diminished an increase in the percentage of CD3+, Thy 1.2+ and TCR alpha beta+ lymphocytes. PMID- 8239521 TI - Variations in temperature and oxygen content do not alter levels of thiobarbiturate reactive material in human breast tumour cells (ZR-75-1) incubated with gamma-linolenic acid. AB - The mechanism by which tumour cells may be killed in vitro by exogenous polyunsaturated fatty acids may involve lipid peroxidation. Gamma-linolenic acid caused a dose and time-dependent reduction in ZR-75-1 cell growth. However, altering either the incubator temperature (35, 37 and 39 degrees C) or the oxygen content (16, 21 and 26%) had little effect on either the growth of cells in the presence of gamma-linolenic acid or on thiobarbiturate reactive material levels over a 7 day period. Thus, small changes in cell culture conditions do not affect 18:3n-6 cytotoxicity or markers of lipid peroxidation. PMID- 8239522 TI - Depletion of serum methionine by methioninase in mice. AB - Methionine dependence is a tumor-specific metabolic defect found in human cancer cell lines as well as in fresh human tumor specimens. Methionine dependent tumors cease growing when deprived of methionine, unlike normal cells which can substitute homocysteine for methionine for their growth requirement. We have previously purified a stable, endotoxin-free methioninase from the bacterium, Pseudomonas putida. We demonstrate in this report that purified methioninase can lower the serum levels of methionine in normal and nude mice from 60 microM to approximately 5 microM within 1 hour. The circulating half-life of methioninase is approximately 100 minutes in mice after i.v. injection. The enzyme therefore seems to be a good candidate as an antitumor agent for methionine-dependent tumors. PMID- 8239523 TI - Inhibition of human glioma cell proliferation and glutathione S-transferase by ascorbyl esters and interferon. AB - The in vitro effect of ascorbyl esters (ascorbyl-stearate [As-S] and -palmitate [As-P]) and interferon (recombinant human interferon-a2b [rHuIFN-a2b]) on human glioma (U-373) cell proliferation, viability and glutathione-S-transferase (GST) activity was studied. The effect of As-S, As-P and rHuIFN-a2b on cell proliferation and viability was evaluated by [3H] Thymidine incorporation and colorimetric MTT assays, respectively. Incubation of glioma cells with As-S, As-P or rHuIFN-a2b for 24 h resulted in a dose dependent inhibition of cell proliferation (IC50 = 68.0 microM As-S, 86.0 microM As-P and 47.3 Units/ml rHuIFN a2b), and moderate decrease of cell viability. It was found that As-S was a more efficient inhibitor of cell proliferation, viability and GST activity than As-P. GST from U-373 cells was purified. The activity of purified GST towards 1-chloro 2,4-dinitrobenzene (CDNB) was inhibited in a dose dependent manner by ascorbyl esters (I-50 = 27.5 microM As-S and 56.0 microM As-P) but not by rHuIFN-a2b. GST activity of cytosol isolated from U-373 cells which were previously treated with As-S (150 microM) or rHuIFN-a2b (150 units/ml) for 0, 2, 5, 10, 20 and 30 min was sharply decreased during 5 to 10 min of treatment and increased at longer durations of treatment. PMID- 8239524 TI - Hematotoxicity of 5-fluorouracil and alpha-interferon on the human normal hemopoietic progenitors. AB - The toxic effects of a combination of 5 fluorouracil (5-FU) and interferon alpha on normal human hemopoietic progenitors were assessed in vitro. CFU-GM growth was evaluated following 1 h incubation of bone marrow cell suspension with 5-FU 100 micrograms/ml or IFN-alpha 2a 1000 U/ml, or with both drugs, either simultaneously or sequentially added. IFN-alpha did not show toxic effects on normal human hemopoietic progenitors and the severe toxicity of 5-FU on the latter was not modified by IFN-alpha. These results suggest that the changes observed in vivo of 5-FU toxicity induced by IFN-alpha should be attributed to indirect effects of the latter. PMID- 8239525 TI - Similarity of serum-tumor pharmacokinetics of antitumor agents in man and nude mice. AB - A pharmacokinetic comparison was made between nude mice and human gastric cancer patients. This comparison is important in order to optimize the human tumor xenograft-nude mouse system as a screening panel for potential antitumor agents. In this report, mitomycin C (MMC), doxorubicin (DXR), 5-fluorouracil (5-FU) and cisplatin (DDP) were administered to nude mice bearing human tumor subcutaneous xenografts in maximum tolerated doses and to patients with gastric cancer at conventional doses. The concentrations of antitumor agents in serum and tumor were detected by bioassay for MMC and 5-FU, by high performance liquid chromatography for DXR, and by atomic absorption method for DDP. Peak drug concentrations in the serum (Cmax) the mice and humans correlated well with statistical significance (R = 0.999, P < 0.0001). When Cmax and drug concentrations in the tumor (T) the mice and human were compared with each other to evaluate the uptake of drugs into the tumor from the serum and calculated as T/Cmax, similar results were observed for the same agent with statistical significance (r = 0.990, p < 0.02). These results indicate that the human tumor xenograft-nude mouse system and humans are essentially similar pharmacodynamically, which further validates the uses of this system to evaluate potential antitumor agents. PMID- 8239526 TI - Optimization of RNA and DNA purification procedures from human colorectal carcinomas. AB - We optimized procedures to extract RNA and DNA separately from colorectal carcinomas pulverized by Microdismembrator II. Our procedures make it possible to achieve good yields and high degree of purification of both types of macromolecules without affecting their structural integrity. PMID- 8239527 TI - Risk factors of lung cancer by histological category in Taiwan. AB - The relationship between various risk factors and lung cancer by different histological types was evaluated in a case-control study. A total of 72 adenocarcinoma patients and 59 squamous/small cell lung cancer patients, 262 hospital controls and 262 neighborhood controls were interviewed. Multiple conditional logistic regression analyses revealed that occupational exposures to asbestos and working as a cook were significant risk factors associated with adenocarcinoma of the lung. An inverse association between incense burning and the adenocarcinoma was noted. The squamous and small cell carcinomas of the lung were significantly associated with cigarette smoking, passive smoking exposure from friends at entertainment activities, the use of coal as cooking fuel, history of prior tuberculosis and chronic bronchitis, and occupational exposures to asbestos. PMID- 8239528 TI - Anti-proliferative effect of combined tumour necrosis factor-alpha and interferon alpha on human pancreatic cancer in vitro and in vivo. AB - The anti-proliferative effects of tumour necrosis factor-alpha (TNF-alpha) and interferon-alpha (IFN-alpha), alone or in combination, on human pancreatic cancer cells lines (PANC-1, MIA PaCa-2 and BxPC-3) and human pancreatic cancer tumour (Exp-58), were investigated in vitro and in vivo. The anti-proliferative effect was determined using the dye uptake method and the subcutaneous tumour model. Combined TNF-alpha and IFN-alpha demonstrated marked synergistic and/or additive effects in comparison with their effects as single agents. These results suggest that combined cytokine therapy of TNF-alpha and IFN-alpha may make possible some improvement in the treatment of pancreatic carcinoma patients in the future. PMID- 8239529 TI - Cis-diamminedichloroplatinum (II)-procaine pharmacokinetic interaction in mice bearing P388 leukemia. AB - The distribution and elimination kinetics of cis-diamminedichloroplatinum (II) (DDP) in female BDF1 mice bearing 6-day P388 leukemia were investigated in the presence and absence of procaine hydrochloride (P.HCl) exposure. DDP was administered as a single i.p. dose of 8 mg/kg in a 0.9% NaCl solution 6 days after tumor inoculum. P.HCl was administered as a single i.v. dose of 40 mg/kg immediately after DDP. The combined treatment with P.HCl produced marked changes in the plasma concentration-time profile of Pt. The unbound fraction of Pt was significantly increased both in the ascites fluid and plasma following DDP + P.HCl administration. P.HCl treatment induced a significant reduction (P < 0.01) in the rate constant of the protein-bound of Pt in plasma of tumored mice. Urinary excretion of Pt was unaffected by P.HCl, and there was no significant P.HCl-induced modification in the concentrations of Pt in the P388 leukemic cells. A statistically significant reduction of kidney and spleen Pt content was observed in female mice exposed to a dose of 8 mg/kg DDP + P.HCl. A similar reduction was observed in kidneys and testes of tumored mice receiving 16 mg/kg DDP along with 40 mg/kg P.HCl, which also showed lower renal and testicular cisplatin-DNA adducts after DDP + P.HCl than after DDP treatment. Potential explanations for the ability of P.HCl to interfere with the pharmacokinetics and biodistribution of DDP are discussed. PMID- 8239530 TI - Effects of dietary menhaden oil on DNA adducts of the food mutagen 2-amino-3 methylimidazo[4,5-f]quinoline (IQ) in Fischer-344 rats. AB - 2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) is a food mutagen which has been found to be carcinogenic in the Fischer-344 rat. We have assessed the effects of a diet containing menhaden oil on IQ-DNA adduct formation in several organs of the male Fischer-344 rat. Young adult, male Fischer-344 rats were maintained on either a) an AIN-76A diet, containing 5% corn oil, or b) an isocaloric AIN-76A diet modified to contain 2% corn oil and 19% menhaden oil (MO diet), or c) a regular laboratory rodent diet (chow diet) for six weeks prior to receiving a single p.o. dose of 10 or 50 mg IQ/kg. Groups of four animals were killed one or six days after IQ administration. Using 32P-postlabeling assays, IQ-DNA adducts were isolated and quantitated in the liver, lungs, stomach, small intestine, large intestine, kidney, heart, spleen and white blood cells. Adduct patterns were similar in all cases and adduct levels, expressed as relative adduct labeling values (RAL x 10(7), were related to dose in all organs. Adduct formation was dependent on the diet and in almost all organs adduct levels in animals on the AIN-76A diet were similar to those on the chow diet. The 19% MO diet inhibited adduct formation in several organs, but not in others. On day 6 there was evidence of impairment of adduct removal in a number of organs of animals on the 19% MO diet and in several organs of animals on the AIN-76A diet, but not in animals on the chow diet. While dietary MO is generally thought to inhibit experimental tumorigenesis at the promotional stage, the present results show that MO diets may also inhibit the initiation process by inhibiting the initial rate of IQ-DNA adduct formation. It also appears, however, that MO diets may impair the process of adduct removal. PMID- 8239531 TI - Effects of the polyamine analogs BE-3-7-3, 3-8-3, and BE-3-8-3 on human brain tumor cell growth and survival. AB - Based on computer modeling, physicochemical studies of spermine-DNA interactions, and cell culture experiments, we hypothesized that polyamine analogs with hydrocarbon chain lengths differing from natural polyamines and a stronger affinity for nucleic acids than spermine should have a cellular antiproliferative effect. We tested three spermine analogs with long hydrocarbon chains, 1,16 diamino-4,12-diazahexadecane (3-8-3), 1,16-bis(ethyl)amino-4,12-diazahexadecane (BE-3-8-3), and 1,15-bis(ethyl)amino-4,11-diazapentadecane (BE-3-7-3) in human brain tumor cell lines U-251 MG, SF-126, and SF-188. Analog concentrations < or = 5 microM inhibited growth and colony-forming efficiency in each cell line by treatment day 5, with significant decreases in putrescine and spermidine, but not spermine, levels. These findings suggest that potentially cytotoxic polyamine analogs can be specified on the basis of their hydrocarbon chain length and DNA affinity. PMID- 8239532 TI - DNA breakage, cytotoxicity, drug accumulation and retention in two human ovarian tumor cell lines AZ224 and AZ364 treated with adriamycin, modulated by verapamil. AB - We investigated the cytotoxicity of adriamycin in two human ovarian tumor cell lines, AZ224 and AZ364, by the MTT-test and we analysed the formation of DNA single-strand (SSB) and double-strand breaks (DSB) by means of the alkaline and neutral elution technique. The AZ364 cell line was 15 times more resistant to ADR (ID50 = 10.0 micrograms/ml) than the AZ224 cell line (ID50 = 0.66 microgram/ml) after 1hr of drug exposure. Immediately after exposure, we observed a biphasic dose response for SSB in the AZ224 cells over a concentration range of 0.1 to 32.0 micrograms/ml, while practically no DSB were found. Upon drug removal and incubation in drug-free medium, full repair of SSB was observed for an ADR concentration of 1 microgram/ml. On the contrary, the DSB became significantly increased for all tested concentrations and persisted as observed after 3hr and 7hr in drug-free medium. The resistant cell line AZ 364 showed consistently less DNA breakage than the AZ 224 cell line. This inherent difference in sensitivity to ADR could, however, not be explained on the basis of the cellular pharmacokinetics of the drug. Verapamil induced a 3 to 4 fold potentiation of the ADR cytotoxicity in both cell lines after continuous exposure and was associated with an increase in DNA-breakage. The results of our study confirm that there is a lack of correlation between cytotoxicity of ADR and DNA strand breakage immediately after 1 hr of drug exposure. Instead, we emphasize the importance of the formation, extent and persistence of protein-associated DSB upon drug removal to the cytotoxic action of ADR in vitro. PMID- 8239533 TI - Interrelation between Western type cancers and non-Western type cancers as regards their risk variations in time and space. V. Mathematical analysis of the environment-sensing signal transmission system in carcinogenesis. AB - The purpose of this study was to elucidate the nature of the control mechanism of human carcinogenesis by applying regression analysis technique to the world statistics of various human neoplasias. The information used in the present study covers 19 cancers in the tumor spectrum, 47 cancer registration areas in space, and the years 1978 to 1982 in time. The risk for each cancer was expressed in terms of the annual age-adjusted incidence rate (AAIR). Results obtained are as follows: 1) The regression analysis of the world statistics demonstrated the existence of 3 subfamilies of cancer: a) Western type subfamily including 13 cancers; b) non-Western type subfamily including 4 cancers; c) independent type subfamily including 2 cancers. 2) Further regression analysis of the world statistics data revealed the presence of a mathematical regularity that characterized the relationship between various cancers and their risk changes in 47 areas of the world. The mathematical regularity of the above intercancer linkage closely resembled that of intersteroidal linkage of urine that was derived from the law of mass action. On the basis of the calculation results with the intercancer linkage, the international risk differences of various cancers are explained by use of the equilibrium model that resembles that of the chemical equilibrium model. Evidence is also presented to suggest that the intercancer equilibrium is maintained through the interaction between a proto-oncogen and a tumor suppressor gene in carcinogenesis. The significance of the environment dependent control mechanism of human carcinogenesis is discussed in the light of population ecology. PMID- 8239534 TI - A carbonic anhydrase inhibitor as a potential modulator of cancer therapies. AB - Since several anticancer drugs are known to become more cytotoxic to cells in an acidic milieu, we have attempted to utilize the carbonic anhydrase inhibitor, Acetazolamide, to acidify the blood and tumor of C3H mice bearing the FSaIIC fibrosarcoma in order to sensitize tumor cells in vivo to CDDP, Melphalan, BCNU, SR4233 or PtCl4 (Fast Black)2 +/- hyperthermia. The direct cytotoxic interactions between the anticancer drugs and Acetazolamide were tested in FSaIIC cells in vitro with the monacidifying diuretic Chlorothrozide as a control. When cells were exposed to CDDP both diuretics protected against cytotoxicity in a dose dependent fashion. In contrast, cells exposed to Melphalan were minimally sensitized and those exposed to BCNU, SR4233, or PtCl4 (Fast Black)2 were essentially unaffected by the presence of the diuretic agents. Both diuretics were essentially non-toxic to cells in vitro, and, interestingly, both drugs markedly protected cells against hyperthermia under low pH conditions. In vitro, however, Acetzolamide produced a tumor growth delay of 2.3 days alone when given at 10 mg/kg i.p. once (the most effective dose) and produced additive growth delays with CDDP and Melphalan, but probably greater than additive delays with SR4233 and PtCl4 (Fast Black)2. When Acetazolamide was given daily for 5 days starting on the day the anticancer drugs were given once (day 7) essentially no further increase in tumor growth delay of nearly 16 days was observed versus only 4,6 days for the drug alone. When hyperthermia (43 degrees C min.) was delivered locally to the tumor after i.p. injection of the drugs, further growth delays were produced for every drug combination which probably were additive in extent. Blood and urine pH determinations revealed that a pH drop of 1 units occurred in the blood and a pH elevation of 1 to 21 units occurred in the urine 1 hr. after i.p. injection of Acetazolamide. These results indicate that this carbonic anhydrase inhibitor can add to the anticancer activity of the drugs tested. The mechanism may involve its ability to acidify the intratumoral environment, but other mechanisms can not be excluded. PMID- 8239535 TI - Detection of human leukemia cells with multidrug-resistance phenotype using multilabeling with fluorescent dyes. AB - Reduced accumulation of multiple drugs is a characteristic of cells overexpressing P-glycoprotein. This phenotype is referred to as multidrug resistance (MDR). A protocol based on reduced accumulation of fluorescent dyes is proposed for discriminating MDR cells in cell populations. The combination of three fluorescent dyes, Hoechst 33342, rhodamine 123 and Nile red, with different intracellular targets, has been designed to characterize cells with different levels of resistance, using image cytometry. The fluorescence intensity of each dye was quantified in living cells. The protocol was applied to human leukemia cell lines, (K562, K562/ADR, CCRF-CEM, CEM/VLB100, CEM/VM-1). The effect of verapamil on dye accumulation is emphasized. PMID- 8239536 TI - Retrovirus-mediated gene transfer of tumor necrosis factor alpha into colon carcinoma cells generates a growth inhibition. AB - Retrovirus-mediated gene transfer was employed to introduce two different cDNAs of the human tumor necrosis factor alpha gene (TNF) into colon carcinoma cell lines either sensitive (LoVo) or resistant (LS174T) to the external addition of TNF. The TNF variants differed in their 5'-sequences (HTNF: authentic TNF cDNA, SPTNF: TNF cDNA with a signal sequence of the IgG heavy chain). The integration of TNF-specific DNA into the genomic DNA, the expression of TNF-specific mRNA and the translation into TNF protein was shown by Southern-, Northern- and Western analysis with TNF-specific DNA- and RNA probes and anti-TNF-antibodies, respectively. Neomycin selected tumor cell clones expressed biologically active TNF up to 40 pg/ml. As shown in a growth assay, both variants of TNF caused an up to 80% growth inhibition in colon carcinoma cells. Furthermore, the cells constitutively producing TNF had a distinguishable morphological appearance and a reduced colony growth in soft agar compared to the parental cells. The subcutaneous injection of 1 x 10(6) TNF-expressing tumor cells into nude mice resulted in a reduced tumor progression in comparison to the controls. PMID- 8239537 TI - The third zinc finger of the WT1 gene is mutated in Wilms' tumour but not in a broad range of other urogenital tumours. AB - Aberrations in the WT1 tumour suppressor gene have been documented in a fraction of Wilms' tumours (WTs). Encoding a protein with four zinc fingers, the WT1 gene is expressed in the developing kidney, gonads, uterus, spleen, mesothelium and brain. Using polymerase chain reaction (PCR), single strand conformation polymorphism (SSCP) analysis and direct DNA sequencing, we analysed 156 diverse tumours for abnormalities of zinc finger 3 (ZF3), a mutational hotspot in WT1. Only one sample (WT) exhibited PCR-SSCP mobility shift. A CGA to TGA nonsense mutation at codon 390 with arginine being substituted with a stop codon was detected and predicted to encode a faulty WT1 protein in this WT, out of 8 WTs studied. Our results are consistent with the presence of WT1 ZF3 mutations in a subset of WTs, but not in other tumours of urogenital nor of WT1-related origin. PMID- 8239538 TI - Frequency of micronuclei in lymphocytes following gamma and fast-neutron irradiations. AB - The dose response of the number of micronuclei in cytokinesis-blocked (CB) lymphocytes after in vitro irradiation with y-rays and neutrons in the 5 dose range was studied in a heterogeneous population of 4 donors. One thousand binucleated cells were systematically scored for micronuclei. Measurements performed after irradiation showed a dose-related increase in micronuclei (MN) frequency in each of the donors studied. The dose-response curves were analyzed by a linear-quadratic model; frequencies per 1000 CB cells were (0.31 +/- 0.049)D + (0.0022 +/- 0.0002)D2 + (13.19 +/- 1.854) (r2 = 1.000, x2 = 0.7074, p = 0.95) following y-irradiation, and (0.99 +/- 0.528)D + (0.0093 +/- 0.0047)D2 + (13.31 +/- 7.309) (r2 = 0.996, x2 = 7.6834, p = 0.11) following neutron irradiation (D is irradiation dose in cGy). The relative biological effectiveness (RBE) of neutrons compared with y-rays was estimated by best fitting linear-quadratic model. In the micronuclei frequency between 0.05 and 0.8 per cell, the RBE of neutrons was 2.37 +/- 0.17. Since the MN assay is simple and rapid, it may be a good tool for evaluating the y-ray and neutron response. PMID- 8239539 TI - Chemoresistant cell lines: morphonuclear characteristics and chemosensitivity development during long-term culture. AB - The aims of the present work were first to study the stability of the sensitivity and resistance of various cancer cell lines over the period of one year, and second to attempt to correlate the morphological aspects of the lines with the results obtained. To this end we used one mammary and two human vesical lines, each consisting of three different phenotypes: a primitive phenotype (the sensitive line); a phenotype able to proliferate in the presence of three antineoplastic agents (the resistant line); and a phenotype derived from the resistant line but cultivated in the absence of drugs for more than 50 passages. We studied the stability of these lines not only in terms of their chemosensitivity to drugs, but also from the point of view of the morphological characteristics of their nuclei as described by digital image analysis. The results obtained show that the resistant lines retained their resistance even after treatment by antineoplastic agents had been discontinued. However, in this latter case variations became noticeable in their sensitivity to drugs, variations which shifted the line towards either the sensitive or the resistant phenotype. Results obtained from image analysis show that, for each of the lines studied, the morphological aspects of the continuously treated resistant and sensitive lines were phenotype-specific. However, the resistant lines no longer cultivated in the presence of anti-cancer agents exhibited morphonuclear characteristics varying across those of the other two phenotypes. We attribute these variations to modifications in the quality of the serum used. PMID- 8239540 TI - A generalized "tumor growth delay" assay for quantifying alterations of tumor regrowth patterns. AB - Tumor growth delay (TGD) is a widely accepted method for in situ assessment of tumor treatment modalities. However, it has some drawbacks: (1) dependency on arbitrary endpoints and, even more serious, (2) lack of alterations of the tumor growth pattern after treatment. In the present study a generalization of TGD is proposed which is based on the analysis of tumor growth curves by applying the difference equation of the Gompertz function. The advantages of the present method consist primarily of addressing the drawbacks of the classical assay mentioned above: generalized TGD permits a quantitative evaluation of treatment induced alterations of tumor-growth patterns after treatment as well as estimations of TGD in a classical form by comparing growth curves as entities, i.e. independently of endpoints. The possibility of studying dose-effect relationships of antineoplastic treatment modalities by the generalized TGD is demonstrated by applying this method on experimental chemotherapy and radiotherapy data. PMID- 8239541 TI - The spin trap alpha-phenyl-tert-butyl nitrone protects against myelotoxicity and cardiotoxicity of adriamycin while preserving the cytotoxic activity. AB - The possibility of preventing the myelo- and cardiotoxicity of adriamycin (ADR), was explored in in vivo experiments in the rat. Assuming that free radicals play a key role in the ADR organotoxicity, a new anti-radical approach was set up by administering a spin trapping compound (PBN) which is taken up by the cells and specifically interacts with radicals. ADR was given i.v., 3 mg/kg every 3rd day for 3 times. PBN was continuously administered throughout the persistence time of ADR in myocardium (15 days). Serial ECG and leukocyte counting were performed during 10 weeks, then hearts were isolated and Langendorff-perfused; functional parameters (heart rate, contractility, coronary flow) were evaluated. PBN improved ECG and prevented the myelotoxicity, while functional parameters were not significantly different from those of control. Cytotoxicity was evaluated in vitro in 3 different human tumour cell lines; PBN did not modify the cytotoxicity of ADR, thus excluding a free radical involvement in this activity. The present results suggest that a proper administration schedule of spin traps might be a promising approach for improving the therapeutic index of ADR. PMID- 8239542 TI - Effect of DNA topoisomerase I inhibitor, 10-hydroxycamptothecin, on the structure and function of nuclei and nuclear matrix in bladder carcinoma MBT-2 cells. AB - The effects of 10-hydroxycamptothecin (HCPT), a DNA topoisomerase I inhibitor, on the structure of nuclei and nuclear matrix and new DNA replication was investigated in murine bladder carcinoma MBT-2 cells. Following 10-30 min exposure of exponentially growing cells to 10-50 microM HCPT, the rate of 2 min pulse-labeling with [3H]thymidine in whole nuclei, nonmatrix, and matrix fractions was markedly decreased in a concentration- and time-dependent manner. Reduction was highest in the nuclear matrix. After treatment of cultures with 50 microM HCPT for 10 min, inhibition of DNA synthesis was 60% in the nuclear matrix and 27% in the low-salt extract fraction tended to be stimulated after drug treatment. SDS-PAGE analysis of [35S]methionine labeled proteins indicated that 50 microM HCPT decreased protein synthesis in the matrix and other nuclear subfractions. HCPT-induced ultrastructural changes in nuclei and nuclear matrix were similar to those typically associated with lesions of DNA replication or RNA transcription. PMID- 8239543 TI - p53 gene mutation spectrum in human unknown primary tumors. AB - Mutations affecting the p53 gene are associated with many human malignancies, but little is known about changes in p53 in unknown primary tumors (UPTs), which are characterized as tumors with advanced stages of malignancy. We therefore investigated the frequency of p53 mutations in a series of 15 unknown primary tumor biopsies and eight cell lines established from UPTs. Mutations in the conserved regions of the p53 gene were verified by single-strand conformation polymorphism analysis of exons 5-9 and were verified by direct DNA sequencing of polymerase chain reaction products. A point mutation leading to an amino acid change in the p53 protein was found in six cases, and a mutation causing a change to termination was found in one case. A frameshift mutation was observed in one cell line. In one patient and one cell line we observed more than one mutation in the p53 coding sequence. Overall, the frequency of mutations that changed the p53 coding sequence in the UPTs we studied was 26% (6/23). Mutations were distributed in eight codons of the p53 gene. Seven of these tumors showed a reduction to homozygosity at the p53 allele, but one tumor apparently retained heterozygosity. We conclude that although UPTs represent highly metastatic advanced tumors that are expected to have a high incidence of p53 mutations, the frequency of p53 mutations is relatively low, suggesting that p53 mutations may not play a major role in the development and progression of this unique tumor type. PMID- 8239544 TI - Influence of cAMP on E-cadherin expression and cell surface heparan sulfate proteoglycan synthesis in human breast cancer cells. AB - The growth of MCF-7 and MDA-MB-231 human breast cancer cells was inhibited by treatment with dibutyryl cAMP (dBcAMP, 10(-4) M). The effects on E-cadherin expression and cell surface associated heparan sulfate proteoglycans (HSPG) synthesis, both implicated in cell adhesion, were investigated. dBcAMP was demonstrated to increase E-cadherin expression in the E-cadherin positive MCF-7 cells. However, in the E-cadherin negative MDA-MB-231 cells, the treatment did not induce expression of this cell adhesion molecule. Furthermore, in the two cell lines, an increase of the [35S] Na2SO4 incorporation into the cell surface sulfated PG was observed subsequently to dBcAMP treatment. Interestingly, the proportion of cell surface HSPG was also enhanced by this treatment. Taken together, these results demonstrate that the decrease of the proliferation observed in the human breast cancer cells after dBcAMP treatment is associated with an increase in the cell-cell and cell-matrix interactions. This suggests that the metastatic process which involves lack of cohesiveness and migration of the cells may probably be counteracted by cAMP in the human breast cancer cells. PMID- 8239545 TI - [General considerations on and the role of internal urethrotomy in the treatment of urethral stricture]. AB - After stressing the importance of a precise diagnosis, the author defines the place of internal urethrotomy, which is certainly not always the first-line treatment that some would claim, due to its poor success rate and because an endoscopic reoperation is sometimes much less harmless than it appears. The importance of perfect anaesthesia for urethral surgery is then emphasised in view of the particular demands of this surgery. Finally, the author reviews the various approaches to the urethra and opening of the stricture. PMID- 8239546 TI - [Principles of wound healing as applied to urethra surgery]. AB - The authors describe the physiology of wound-healing and the factors influencing wound-healing that may play a role in urethral surgery. They present the specific characteristics of wound-healing in each treatment for urethral strictures. This knowledge can help to choose the right treatment for each type of urethral stricture. PMID- 8239547 TI - Two stage scrotal flap urethroplasty. AB - This technique is derived from the Denis Browne two-stage technique for hypospadias. It was developed by Johanson and perfected by Turner-Warwick. It uses an invaginated flap of scrotal skin sutured to the edges of the exposed stenotic urethra. The second stage (2 to 3 months later) closes the urethra in two layers. No statistics are available for the results obtained. PMID- 8239548 TI - [Urethroplasty using a free dermo-epidermal of preputial mesh graft: Meshgraft urethroplasty]. AB - Long urethral strictures raise serious problems in terms of appropriate treatment. Reconstruction techniques using scrotal skin have a high recurrence rate. In order to avoid complications, a dermoepidermal or preputial Meshgraft was used to reconstruct a new urethra in two stages. In the first stage, the graft was raised and transplanted onto the open urethra. 8 to 12 weeks later, in the second stage, the new urethra is closed. Since 1977, Meshgraft urethroplasty has been performed in 128 patients, using the skin of the foreskin (n = 75) or medial surface of the thigh (n = 53). A good anatomical and functional result was obtained in all but three cases, regardless of the graft donor site. This operative technique constitutes a very effective method for the lasting treatment of long or complicated urethral strictures, for example in patients with spinal cord injuries. PMID- 8239549 TI - One-stage transverse distal penile/preputial island flap urethroplasty for urethral stricture. AB - The rich blood supply of the penis, derived from three longitudinal arterial and venous pedicles and numerous transvesical anastomoses allows the raising of a full thickness skin graft obliquely underneath the glans and around the penis. An island flap, which can be as long as 15 cm, is isolated but its blood supply is preserved by cleaving the connective tissue underneath the basic flap. The vascularised island flap is then rotated towards the zone of urethral replacement either in the form of a flap or a tube which can extend as far as the membranous urethra. 150 cases have been operated with this pedicle flap as a one-stage operation. No cases of necrosis were observed, even in the presence of fistulae or periurethral abscesses. A post-operative proximal stricture was observed in 10% of cases and was treated by internal urethrotomy. One third of these failures had to be reoperated with no major difficulties. Two pre-existing urethrocutaneous fistulae also had to be reoperated. PMID- 8239550 TI - [Urethroplasty using a full-thickness free skin graft]. AB - Full-thickness free skin graft urethroplasty is an excellent technique for the cure of urethral strictures. Based on his experience of 250 cases since 1977, the author describes the indications and limitations related to surrounding tissues and to the graft itself. A rigorous and precise technique, described in detail in this paper, is the key to success. PMID- 8239551 TI - [Urethral stenosis in children: treatment using urethroplasty with a vesical or oral mucosal graft]. AB - Urethral strictures are rare in children and correspond to either congenital membranes, post-traumatic (endoscopy). The precise diagnosis is obtained by voiding cystometrography at the end of urography or by retrograde opacification. Congenital membranes should be treated by dilatation and limited strictures should be treated by urethrotomy. The other cases warrant urethroplasty with a free vesical or oral mucosal flap. Good results are obtained in about 90% of cases. PMID- 8239552 TI - [End-to-end anastomosis of the urethra]. AB - End-to-end anastomosis is perhaps the most physiological method to cure a stricture of the urethra, provided it is short and the rest of urethra is elastic enough to overbridge the gap without tension. A success rate of more than 90% can be expected in these circumstances. PMID- 8239553 TI - The use of urethral stents for the treatment of urethral strictures. AB - Three types of urethral stents can be used in the treatment of recurrent strictures: Urolume/Wallstent is a self-expanding mesh which is incorporated into the urethral epithelium; the ASI Titanium stent is a short rigid mesh of Titanium wire which is also incorporated into the urethra; Nitinol is a flexible spring in one or two parts connected by a steel wire. It remains endoluminal. The Urolume/Wallstent has been used since 1987. It is indicated in iatrogenic strictures. No statistical results are presented. The ASI stent should be reserved for strictures of the prostatic and posterior urethra. The Niticol UroCoil system can be used at all levels of the urethra; two models are available: simple or twin. The author has inserted 65 stents in 56 patients. The stent was removed after one year in 41 patients. The use of these stents has considerably decreased the number of repeated dilatations and urethrotomies. PMID- 8239554 TI - [Cancerous stricture of the urethra]. AB - When certain strictures which have been treated for a long time start to bleed, the possibility of carcinoma should be considered. However, cancer may present in the form of more recent and deteriorating dysuria. The diagnosis is based on urethrography which shows a narrow and tortuous pass. Biopsy should be performed under urethroscopic guidance. The cancer is usually in the bulbomembranous segment. Treatment is disappointing. Wide excision is very badly disabling. PMID- 8239555 TI - [Stricture of the urethral meatus and the navicular fossa]. AB - A review of the literature concerning strictures of the meatus and fossa navicularis reveals that few techniques are entirely satisfactory, apart from major techniques which are difficult to perform. The author describes an easy technique which gave perfectly satisfactory results in the majority of 36 cases, 17 of which have a follow-up of more than 5 years. PMID- 8239556 TI - Superficial siderosis. PMID- 8239557 TI - Experimental allergic encephalomyelitis as a guide to the understanding and treatment of multiple sclerosis. PMID- 8239558 TI - The pathogenesis of superficial siderosis of the central nervous system. AB - In advanced cases of superficial siderosis of the human central nervous system, the clinical triad of hearing loss, cerebellar ataxia, and myelopathy permits the diagnosis at the bedside, and magnetic resonance imaging readily confirms the hemosiderin deposits in brainstem, cerebellum, and spinal cord. To study the pathogenesis of this condition and explain the selective vulnerability of the cerebellum, experimental siderosis was induced in rabbits by the repeated intracisternal injection of autologous red blood cells. The earliest cellular response in the cerebellar molecular layer was hyperplasia and hypertrophy of microglia as displayed by immunocytochemistry for ferritin. Microglia also contained iron, but ferritin biosynthesis appeared to proceed without commensurate iron accumulation. This early apoferritin response probably occurred due to the presence of heme, rather than iron, in the cerebrospinal fluid and subpial tissue. Ferritin biosynthesis is accelerated when the ferritin repressor protein is dissociated from ferritin messenger ribonucleic acid. A specific antiserum localized ferritin repressor protein predominantly to astrocytes including Bergmann glia. It is proposed that abundance and proximity of ferritin repressor protein--immunoreactive Bergmann glia and ferritin-containing microglia in the cerebellar molecular layer permit prompt cellular interaction in the conversion of heme to ferritin and ultimately hemosiderin. PMID- 8239559 TI - Inhibition of acute, experimental autoimmune encephalomyelitis by the synthetic immunomodulator linomide. AB - Linomide (LS-2616, quinoline-3-carboxamide) is a synthetic immunomodulator that stimulates natural killer cell activity and activates several lymphocytic subpopulations in experimental animals and humans. In this study we determined the effect of oral treatment with linomide on the development of experimental autoimmune encephalomyelitis, an animal model for immune-mediated human demyelinating disorders. Experimental autoimmune encephalomyelitis was induced in SJL/J mice and in an outbred strain of rats (Sabra) by subcutaneous injection of spinal cord homogenate in adjuvant followed by inoculation with Bordetella pertussis. Linomide was administered in drinking water, at an estimated dose of 50 to 100 mg/kg/day. None of the linomide-treated mice (0/41) and Sabra rats (0/15) developed any clinical or pathological signs of experimental autoimmune encephalomyelitis, whereas almost all control animals (48/53 and 18/19, respectively) were severely paralyzed and 64.5% died from the disease. Lymphocytes obtained from linomide-treated animals had reduced in vitro proliferative responses to guinea pig myelin basic protein, proteolipid protein of the myelin, and tuberculin-purified protein derivative, unlike antigen independent proliferation which was rather unaffected. Natural killer cell activity (tested by a cytotoxic assay on radiolabeled YAC-1 target cells) was significantly enhanced in mice treated with linomide. Our results indicate that modulation of the immune system with linomide leads to complete inhibition of experimental autoimmune encephalomyelitis in the absence of systemic immunosuppression. Linomide could therefore be of use in future clinical trials for the treatment of human autoimmune demyelinating disorders. PMID- 8239560 TI - Correlation between magnetic resonance imaging findings and lesion development in chronic, active multiple sclerosis. AB - Magnetic resonance imaging is a highly sensitive method for the detection of the lesions of multiple sclerosis and renders possible the study and the evolution of early lesions. Previous reports on magnetic resonance imaging following gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) injection demonstrated that new lesions can be recognized by contrast enhancement. The pathological basis of these observations is uncertain. We have had the opportunity to study at autopsy the brain of a patient with chronic progressive multiple sclerosis who suffered acute worsening leading to death. Magnetic resonance imaging performed 10 days and 4 weeks prior to death showed new Gd-DTPA-enhanced lesions in the posterior hemispheric white matter adjacent to the lateral ventricles. Light microscopic examination of these areas demonstrated them to be fresh lesions comprising intense inflammatory activity and dense perivascular cuffs within an edematous lesion center and a striking parenchymal mononuclear cell infiltration at the margins of the lesions. Lesions that were demonstrated by increased signal on T2-weighted images, but were not enhanced following administration of Gd-DTPA, were all of the chronic type, either inactive or active. None of these showed the intense inflammatory activity of the acute lesions and most displayed fibrous astrogliosis. PMID- 8239561 TI - Spastic paraparesis associated with human T-lymphotropic virus type I: a clinical, serological, and genomic study in Iranian-born Mashhadi Jews. AB - The Mashhadi-Jewish community originating in Iran is a closed and ethnically segregated population with a unique history and a high rate of intrafamilial marriage among its members. A high risk of infection by human T-lymphotropic virus type I (HTLV-I) and of adult T-cell leukemia associated with such infection was found in this population. HTLV-I is also associated with a syndrome of progressive spastic paraparesis. We therefore evaluated the occurrence of HTLV-I infection and spastic paraparesis in Mashhadi-born Iranian Jews who immigrated to Israel. We examined 83 Mashhadi-born subjects (52 women, 31 men; mean age, 61 +/- 15.5 years) and 73 age-matched non-Mashhadi Iranian-born Jews. Blood samples were tested for HTLV-I antibodies by particle agglutination test. The polymerase chain reaction (PCR) was used to detect HTLV-I proviral DNA sequences from peripheral blood mononuclear cells. Fifteen Mashhadi-born Jews (18%) were both seropositive and PCR-positive for HTLV-I. Four HTLV-I-seronegative subjects were found to be positive for HTLV-I proviral DNA by PCR. Of the 19 HTLV-I-infected subjects (11 women, 8 men; mean age, 59 +/- 16 years), 13 (68%) had spastic paraparesis of varying severity. There were no signs of myelopathy in the Mashhadi-born subjects who were negative for HTLV-I proviral DNA by PCR. None of the non-Mashhadi Iranian Jews was seropositive or PCR-positive for HTLV-I proviral DNA, or had clinical signs of spastic paraparesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239562 TI - Forearm 3-methylhistidine efflux in myotonic dystrophy. AB - Myotonic dystrophy is associated with progressive muscular atrophy. To define the mechanism of muscle wasting in this disease, we studied myofibrillar proteolysis in vivo in 8 men moderately affected with myotonic dystrophy, and compared the results with those of 10 normal men. Myofibrillar proteolysis was estimated by measuring the 3-methylhistidine arteriovenous difference (A-V) and efflux (Q) across the forearm in the postabsorptive state. Plasma 3-methylhistidine concentrations were determined by high-performance liquid chromatography with postcolumn o-phthalaldehyde derivatization and fluorescence detection. Plasma flow to the forearm muscles (F) was estimated to represent 85% of total forearm plasma flow as determined by the indicator-dilution technique. Forearm 3 methylhistidine efflux was calculated as: Q = F(A-V). Mean muscle mass (24-hour creatinine excretion), lean body mass, and forearm volume were decreased in the patients with myotonic dystrophy, confirming the presence of muscle atrophy. Mean forearm 3-methylhistidine arteriovenous difference and efflux were not significantly different in the two groups. We conclude that myofibrillar protein degradation is not increased in myotonic dystrophy, even when measured in a muscle compartment selectively affected by wasting. Muscle atrophy in myotonic dystrophy is probably the result of defective anabolism rather than accelerated catabolism. PMID- 8239563 TI - Disequilibrium of unknown cause in older people. AB - Many older people complain of a sense of unsteadiness when standing or walking and many are fearful of falling. The cause of this disequilibrium is often difficult to determine even after thorough clinical evaluation. To better understand the pathogenesis of this symptom in a community-dwelling population, we studied 26 patients > 75 years old who complained of disequilibrium in whom no cause was evident after clinical evaluation. Each patient and each of 26 age matched controls underwent a comprehensive evaluation that included a history and examination, a questionnaire, functional scales, a gait and balance scale, mini mental state testing, audiometry, visual acuity, visual tracking, rotational vestibular testing, and quantitative posturography. Although none had Romberg's sign, patients tended to sway more and do poorer on semiquantitative gait and balance testing than did controls. Patients showed few differences from controls for most other measurements. However, 7 patients, designated Group V, had profoundly reduced vestibular function. In the remaining 19 patients (Group D), the average vestibular function was still significantly lower than controls. Sway velocity measured with posturography was increased in the patients compared with controls, particularly in Group V patients. Measures of vestibular function correlated with increased sway velocity both in Group D alone and among all study participants. Quantitative measurement of vestibular function should be considered in older people complaining of disequilibrium, particularly if the cause is not apparent after the initial evaluation. PMID- 8239564 TI - Interictal indices of temporal seizure origin. AB - Two studies assessed the value of temporal lobe interictal electroencephalographic (EEG) spikes and delta in indicating side of temporal epileptogenesis. The first study determined laterality of spikes/delta in awake recordings of 56 patients whose seizures all began unilaterally as proven by (1) EEG-recorded seizures and (2) > 90% improvement after lobectomy. Spikes of 52 (93%) and delta of 46 (82%) patients predominated or appeared exclusively ipsilateral to seizure origin. Neither predominated contralaterally in any patient. The second study investigated laterality of temporal seizures in a separate group of 156 patients with various side vs side spike or delta ratios on 1 to > or = 4 awake recordings. Ninety-nine of 104 patients (95%) with temporal spikes on four or more awake recordings had most or all seizures ipsilateral to most spikes, including 79 of 80 (99%) of those with > or = 3 side vs side spike ratios. Among the 120 patients with high (> or = 3) side vs side spike ratios, most or all seizures of 118 (98%) originated ipsilateral to most spikes. Predominant seizure origin also correlated with lateralized arrhythmic delta- from 90% ipsilateral seizures of those with one EEG with delta to 100% with > or = 4 such EEGs. Data from these two studies using opposite directions of analysis (seizures-->spikes/delta and spikes/delta-->seizures) demonstrate high correlations between laterality of interictal and ictal entities, particularly if temporal spikes clearly predominate on one side and if unilateral temporal delta activity persists over several recordings. Such correlations suggest that the awake interictal scalp EEG cannot be ignored when assessing laterality of temporal epileptogenesis. PMID- 8239565 TI - Polysomnographic sleep measures in Parkinson's disease patients with treatment induced hallucinations. AB - Prior studies of sleep in Parkinson's disease (PD) have been compromised by inadequate comparison groups, mixed medication regimens, and absence of quantitative data collection. This is the first study to compare polysomnographic sleep measures in PD patients on only dopaminergic medications with and without hallucinations. We performed two consecutive nights of polysomnography in 10 nondepressed, nondemented PD patients, 5 with and 5 without hallucinations. All patients were being treated with carbidopa/levodopa and a dopaminergic agonist only. Hallucinators and nonhallucinators were group-matched for age, PD duration, severity, and medication doses. Both groups had abnormal sleep records. In particular, there was a reduction in K-complexes and spindle formation, and the frequent occurrence of motor activation during rapid eye movement (REM) sleep consistent with REM behavior disorder. The hallucinator group had a significantly lower sleep efficiency (0.25 in hallucinators vs 0.61 in nonhallucinators, p = 0.006), a reduced total REM sleep time (mean total REM sleep time, 3 minutes in hallucinators vs 50 in nonhallucinators; p = 0.005), and a reduced REM percentage (mean, 5% in hallucinators vs 20% in nonhallucinators; p = 0.011). This study demonstrates that advanced PD patients treated with dopaminergic agents have abnormal sleep patterns and that those with dopaminergic-induced hallucinations have significantly greater REM aberrations than nonhallucinating PD patients. PMID- 8239566 TI - Chronic levodopa administration alters cerebral mitochondrial respiratory chain activity. AB - Parkinson's disease (PD) is characterized mainly by a loss of nigrostriatal dopamine neurons. Thus far, the actual physiopathology of PD remains uncertain, although recent studies have found decreased activity of complex I, one of the enzymatic units of the mitochondrial respiratory chain, in various tissues of PD patients. Because most, if not all, of PD patients are treated chronically with levodopa, the precursor of dopamine, and because we have shown previously that catecholamines may alter mitochondrial respiration, we assessed the effects of chronic administration of levodopa on complex I activity in rat brain. We found that chronic administration of levodopa, at a dose used in PD patients, caused a significant reduction in complex I activity while it did not affect the activities of complex II, complex IV, and citrate synthase. Reduction in complex I activity correlated well with catecholamine innervation as the reduction was observed mainly in the striatum and substantia nigra and to a lesser extent in the frontal cortex but not in the cerebellum. Moreover, the levodopa-induced decrease of complex I activity was reversible since activities at 1, 3, and 7 days after the last injection showed a progressive return to control values. Incubation of whole brain mitochondria in vitro showed that both levodopa and dopamine inhibit complex I activity in a dose- and time-dependent manner. In contrast, other compounds such as homovanillic acid, 3,4-dihydroxyphenylacetic acid, and 3-O-methyl-dopa were minimally effective. Reduced glutathione, ascorbate, superoxide dismutase, and catalase prevented the effect of levodopa and dopamine on complex I. Various inhibitors of monoamine oxidase also prevented the effect of dopamine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239567 TI - Evidence for a central cholinergic effect of high-dose thiamine. AB - In vitro animal studies have suggested that thiamine is involved in the presynaptic release of acetylcholine. Total thiamine content in cholinergic nerve terminals is comparable with that of acetylcholine, and the phosphorylation state of thiamine changes with release of acetylcholine. Thiamine binds to nicotinic receptors and may exhibit anticholinesterase activity. Based on these observations, we investigated the effects of pharmacological doses of thiamine on the cognitive deficits induced by the anticholinergic scopolamine in healthy young adults using a randomized, double-blind, placebo-controlled, double crossover design. Drug effects were assessed by P3 event-related potential, quantitated electroencephalography, and free recall memory. Conditions included (1) baseline, (2) thiamine 5 gm p.o. and scopolamine 0.007 mg/kg IM, and (3) lactose PO and scopolamine 0.007 mg/kg IM. Thiamine significantly reduced adverse effects of scopolamine on P3 latency, spectral components of electroencephalography, and memory recall. The results are consistent with a cholinomimetic effect of thiamine in the central nervous system. Additional studies are needed to delineate the basic mechanisms and possible therapeutic efficacy of thiamine at pharmacological dosages. PMID- 8239568 TI - Anterior temporal language areas in patients with early onset of temporal lobe epilepsy. AB - Eighteen consecutive patients undergoing dominant temporal lobectomy underwent preoperative cortical stimulation for language localization. Patients with naming deficits on anterior (4.5 cm from the temporal pole) temporal lobe stimulation had earlier seizure onset vs those without such deficits (5.8 yr vs 12.9 yr; p < 0.04). There was a similar trend for reading errors (6.3 yr vs 12.4 yr; p < 0.052). Resections always spared at least 1 cm anterior to any language area. There was no significant difference in postoperative neuropsychological tests between patients with and without anterior language representation. Early onset of dominant temporal lobe seizure foci leads to a more widespread or atypical distribution of language areas. Individual variability should be considered in epilepsy surgery to reduce postoperative language deficits. PMID- 8239569 TI - Positron emission tomographic studies of dopa-responsive dystonia and early-onset idiopathic parkinsonism. AB - There are two major syndromes presenting in the early decades of life with dystonia and parkinsonism: dopa-responsive dystonia (DRD) and early-onset idiopathic parkinsonism (EOIP). DRD presents predominantly in childhood with prominent dystonia and lesser degrees of parkinsonism. EOIP presents before age 40 with parkinsonism (often with associated dystonia). Both disorders are exquisitely sensitive to levodopa, although the long-term prognosis in each appears to be different. Some have suggested, however, that DRD is a form of EOIP. We performed positron emission tomography with 6-fluoro-dopa in 10 patients with DRD and 18 patients with EOIP to study the integrity of their nigrostriatal dopaminergic systems. In DRD, we found normal striatal FD uptake. In contrast, patients with EOIP had reduced striatal FD uptake. We conclude that the patho physiologies of DRD and EOIP are distinct. Although both disorders presumably represent a deficiency of striatal dopamine, the results suggest that in DRD dopa uptake, decarboxylation, and storage mechanisms are intact. This may explain the sustained response of DRD to low doses of levodopa. 6-Fluoro-dopa positron emission tomography distinguishes DRD from EOIP. PMID- 8239570 TI - Unverricht-Lundborg disease: absence of nonallelic genetic heterogeneity. AB - Unverricht-Lundborg disease is a clinically recognizable form of progressive myoclonus epilepsy. Recently, in several families of both Finnish and Mediterranean extraction segregating Unverricht-Lundborg disease, the gene for this disease was linked to the same region of the long arm of chromosome 21. We performed linkage analysis in eight families, including four of neither Baltic nor Mediterranean origin, using a polymorphic (CA)n repeat marker for the human liver-type 6 phosphofructokinase (PFKL) gene, previously mapped to 21q22.3. No recombinations were observed between the disease phenotype and the PFKL marker and a maximum lod score of 5.63 was obtained. These findings confirm tight linkage between PFKL and the gene for Unverricht-Lundborg disease and strongly suggest a lack of nonallelic genetic heterogeneity of the disease. PMID- 8239571 TI - Significance of reactive Lyme serology in multiple sclerosis. AB - Nineteen of 283 consecutive patients evaluated in the Multiple Sclerosis (MS) Comprehensive Care Center had a borderline or positive B. burgdorferi serology. In 8 patients a repeat serology was nonconfirmatory. Cerebrospinal fluid was examined in 10 seropositive patients and showed anti-B. burgdorferi antibodies without intrathecal production in 5 patients. Antibiotic treatment did not prevent subsequent neurological relapses. The finding of reactive Lyme serology in an MS patient with no suggestive features of the infection is unlikely to indicate neurological Lyme disease. PMID- 8239572 TI - Magnetic resonance imaging measurements and clinical changes accompanying transtentorial and foramen magnum brain herniation. PMID- 8239573 TI - High incidence of hepatitis C virus infection in patients with cryoglobulinemic neuropathy. PMID- 8239574 TI - Multifocal motor neuropathy or CIDP? PMID- 8239575 TI - The apolipoprotein epsilon 4 allele in patients with Alzheimer's disease. AB - Apolipoprotein E (APO-E) binds to the beta-amyloid peptide and is present in senile neuritic plaques in Alzheimer's disease (AD). The epsilon 4 isoform of APO E has been associated with both sporadic and familial late-onset AD, implying a causal role. Among patients and control subjects similar in age, gender, and ethnic group from the New York City community of Washington Heights-Inwood, we found that the odds ratio (OR) for AD associated with homozygosity for APO epsilon 4 was 17.9 (95% confidence interval [CI], 4.6-69.8) and that associated with heterozygosity for APO-epsilon 4 was 4.2 (95% CI, 1.8-9.5) compared with persons with other APO-E genotypes. The association was stronger among patients with sporadic disease (OR = 10.3; 95% CI, 3.4-31.1) than among those with a family history of dementia in a first-degree relative (OR = 0.9; 95% CI, 0.1 13.5). The association between APO-epsilon 4 and AD did not differ according to age at onset (< 65 vs > or = 65), but appeared to vary across the 3 ethnic groups investigated (black, Hispanic, and white). Our data confirm the association between AD and APO-epsilon 4 and support the hypothesis that the APO-epsilon 4 allele either confers genetic susceptibility to AD or may be in linkage disequilibrium with another susceptibility locus. Ethnic variability in the allelic frequency of APO-epsilon 4 in the elderly warrants further investigation. PMID- 8239576 TI - Treatment of Q fever. PMID- 8239577 TI - Microbial conversion of avermectins by Saccharopolyspora erythraea: glycosylation at C-4' and C-4''. AB - Avermectins and ivermectins are glycosylated at C-4'' and C-4' by both growing and resting cells of Saccharopolyspora erythraea. The reaction is catalyzed by a glycosyltransferase which is constitutive. The enzyme uses UDP-glucose as the glycosyl donor and avermectin or ivermectin mono- and disaccharides as acceptors. Avermectin and ivermectin aglycones and erythromycin A are not substrates for the enzyme. PMID- 8239578 TI - High rate of erythromycin and clarithromycin resistance among Streptococcus pneumoniae isolates from blood cultures from Providence, R.I. AB - Four (5%) of 81 recent isolates of Streptococcus pneumoniae from the blood of adult patients but no isolates from pediatric patients (n = 51) were resistant (MIC, > or = 1 microgram/ml) to erythromycin. The MICs of clarithromycin were slightly lower than those of erythromycin, but there was complete cross resistance. Routine testing and surveillance are needed to determine whether erythromycin resistance among S. pneumoniae isolates is increasing throughout the United States. PMID- 8239579 TI - In vitro activity of azithromycin (CP-62,993) against Chlamydia trachomatis and Chlamydia pneumoniae. AB - The in vitro susceptibilities of 49 strains of Chlamydia trachomatis and 3 strains of Chlamydia pneumoniae to azithromycin and tetracycline or doxycycline were determined. The MIC of azithromycin ranged from < or = 0.06 to 1.0 micrograms/ml, the MIC of tetracycline ranged from 0.03 to 0.12 micrograms/ml, and the MIC of doxycycline ranged from 0.015 to 0.06 micrograms/ml against C. trachomatis. The MIC ranges for C. pneumoniae were 0.12 to 0.25 micrograms/ml for azithromycin and 0.06 to 0.12 micrograms/ml for tetracycline. All minimal chlamydicidal concentrations were either equal to the MIC or one or two dilutions higher. No strains resistant to these antibiotics were detected. In vitro activity shows that azithromycin is highly active against C. trachomatis and C. pneumoniae. PMID- 8239580 TI - Interaction between biofilms formed by Pseudomonas aeruginosa and clarithromycin. AB - Interactions between bacterial biofilms formed by Pseudomonas aeruginosa and clarithromycin, a macrolide having no anti-P. aeruginosa activity, were investigated. P. aeruginosa incubated for 10 days on membrane filters formed biofilms on the surfaces of the filters. The biofilms were characterized by dense colonizations of bacteria and thick membranous structures that covered the colonies. Treatment of the biofilms with a relatively low concentration of clarithromycin for 5 days resulted in an eradication of the membranous structures. Quantitative analysis of alginate and hexose was done to evaluate the quantity of polysaccharides in or on the biofilms. Treatment of the biofilms with clarithromycin decreased the quantity of alginate and hexose and therefore perhaps the quantity of polysaccharides as well. Eradication of the membranous structures of biofilms, or the decrease in the quantity of polysaccharides, resulted in an increase in the rate of penetration of antibiotics through bacterial biofilms. In vivo therapeutic effects of ofloxacin in the rat infection model, in which the biofilm mode of growth of P. aeruginosa is characteristic, were enhanced by oral coadministration of clarithromycin. It is suggested that clarithromycin eradicated glycocalyx produced by P. aeruginosa, or suppressed the production of glycocalyx, by unknown mechanisms and thereby enhanced the therapeutic efficacies of other antimicrobial agents against infections caused by P. aeruginosa. PMID- 8239581 TI - Dose ranging and fractionation of intravenous ciprofloxacin against Pseudomonas aeruginosa and Staphylococcus aureus in an in vitro model of infection. AB - The effect of dose or dose interval on the pharmacodynamics of simulated high dose intravenous ciprofloxacin therapy on infection due to Pseudomonas aeruginosa and Staphylococcus aureus was studied in an in vitro hollow-fiber model of infection. Simulated doses of 1,200 mg of ciprofloxacin per day as either 400 mg every 8 h or 600 mg every 12 h against P. aeruginosa resulted in selection of ciprofloxacin-resistant bacteria. The results with one test strain that was isolated from a patient prior to administration of intravenous ciprofloxacin demonstrated selection of a gyrA mutant in the model, as had occurred in vivo. A single 1,200-mg dose every 24 h did not select for bacterial resistance; however, breakthrough regrowth of ciprofloxacin-susceptible bacteria occurred. Dosages of 400 or 600 mg of ciprofloxacin every 12 h effectively reduced bacterial counts of one strain each of methicillin-susceptible or -resistant S. aureus, with no bacterial resistance detected at the end of experiment; in contrast, 200 mg every 12 h resulted in bacterial regrowth due to the selection of drug-resistant bacteria. These data show the need for high-dose intravenous ciprofloxacin, particularly with regimens producing high peak levels, for treatment of infections where selection for bacterial resistance is a clinical problem. PMID- 8239582 TI - The time of administration of 3'-azido-3'-deoxythymidine (AZT) determines its host toxicity with possible relevance to AZT chemotherapy. AB - 3'-Azido-3'-deoxythymidine (AZT) is the drug most widely used in the treatment of AIDS. Its major drug-related toxicity is bone marrow suppression, which limits the dose of AZT that can be used. It is essential that AZT be phosphorylated for antiviral effect. We have recently demonstrated that thymidine kinase (TK), the initial enzyme in AZT anabolism, follows a circadian pattern in rat bone marrow. The present study was undertaken to determine whether AZT toxicity is related to the time of its administration and whether the variation in toxicity is correlated with the circadian variation in TK activity. Male Sprague-Dawley rats were housed under standardized conditions of light and dark (lights on 0600 to 1800 and lights off 1800 to 0600) for 4 weeks. The animals were randomly divided into seven groups; six groups were administered AZT by intraperitoneal injection at the same dose of 750 mg/kg of body weight at various times (0400, 0800, 1200, 1600, 2000, and 2400), and one group was used as a control. AZT-related toxic effects, including bone marrow toxicity, differed significantly among the treatment groups, depending on the time of AZT administration (by analysis of variance and Cosinor analysis, P < 0.001). The least toxicity was observed in rats receiving AZT at 1600 (10 h after light onset [10 HALO], in late sleep span) and the greatest toxicity was observed in those injected at 0400 (22 HALO, in late activity span). To verify these results, we administered AZT by intraperitoneal injection at an approximately 50% lethal dose (1,500 mg/kg) to two groups of rats, one at 1200 (6 HALO, in the middle of the sleep span) and the other at 2400 (18 HALO, in the middle of the activity span). AZT lethality was significantly higher in rats receiving AZT at 2400 (18 HALO, in the middle of the activity span). Further statistical analysis demonstrated that the variation in AZT toxicity was correlated with the circadian variation in TK activity in bone marrow of the same species (peak activity at 0400 [22 HALO, in late activity span] and trough activity at 1600 [10 HALO, in late sleep span]), suggesting that the circadian variation in TK activity may be the biochemical basis for the observed circadian variation in AZT toxicity. These results may be useful in the design of improved AZT chemotherapeutic regimens. PMID- 8239583 TI - Penetration of foscarnet into cerebrospinal fluid of AIDS patients. AB - The diffusion of foscarnet into cerebrospinal fluid (CSF) was studied in 27 patients with AIDS. Foscarnet was administered intravenously at various dosages at 12-h intervals. Concentrations in plasma and CSF at the end of foscarnet infusion or 1, 3, 5, 6, and 12 h after infusion were determined by high performance liquid chromatography. Thirty-seven samples were obtained. The median concentration of foscarnet in CSF was 80 mumol/liter (range, 0 to 500 mumol/liter). The CSF foscarnet concentration was greater than the 50% inhibitory concentration for human immunodeficiency virus type 1 and was equal to or greater than the 50% inhibitory concentration for cytomegalovirus in most cases. The penetration of foscarnet into CSF, as expressed by the ratio of the concentration in CSF to the simultaneous concentration in plasma, ranged from 0 to 3.4 (median, 0.27) and was highly correlated with the presence of cells within CSF and the length of foscarnet therapy. Good diffusion of foscarnet in CSF allows evaluation of this drug in central nervous system cytomegalovirus and human immunodeficiency virus infections in patients with AIDS. PMID- 8239584 TI - Alterations in pharmacokinetics and protein binding behavior of cefazolin in endotoxemic rats. AB - The possible alterations in the pharmacokinetics and protein binding behavior of the beta-lactam antibiotic cefazolin (CEZ) were investigated in endotoxemic rats induced by Klebsiella pneumoniae O3 lipopolysaccharide (LPS). LPS (250 micrograms/kg of body weight) was infused for 20 to 30 min 2 h before an intravenous administration of CEZ (20 mg/kg). Significant decreases in systemic clearance and renal clearance of CEZ were observed in LPS-treated rats without any changes in fraction of urinary excretion in unchanged CEZ (> 0.8). The volume of distribution at steady state showed a tendency to increase. The protein binding parameters of CEZ, the binding capacity, and number of binding sites on the albumin molecule were decreased by LPS, whereas the dissociation constant did not change. Significant decreases in systemic and renal clearances for unbound CEZ were observed in LPS-treated rats. The glomerular filtration rate estimated as inulin clearance was also decreased by LPS. The ratio of renal clearance of unbound CEZ to glomerular filtration rate (clearance ratio) dropped to 70% of that in control rats, and the net tubular secretion of CEZ was also dramatically reduced. The present study suggests that LPS has an effect on the pharmacokinetics of CEZ by changes which occur in renal handling and protein binding. PMID- 8239585 TI - In vivo administration of azithromycin affects lymphocyte activity in vitro. AB - A therapeutic dose of azithromycin was administered to test subjects and then the following lymphocyte functions were examined in vitro: proliferative lymphocyte response to stimulation with pokeweed mitogen, levels of immunoglobulins G, A, and M in serum, and the amount of the soluble interleukin 2 receptors in supernatants of mononuclear cell cultures stimulated with phytohemagglutinin and phorbol myristate acetate. The study was performed as a controlled clinical trial comparing an azithromycin-treated group (n = 21) and a placebo-treated control group (n = 10). Healthy female volunteers were placed into one of the two groups, and the study was performed as a double-blind trial. Although the findings of the present study showed that azithromycin significantly increased the proliferative lymphocyte response to pokeweed mitogen, the results could have been due to experimental variation. However, impairment of the lymphocyte function was not observed, which could represent valuable information. Likewise, no effect of azithromycin on levels of the immunoglobulins in serum was observed. The most marked effect of azithromycin on the lymphocyte function was demonstrated by an elevation in the amount of soluble interleukin 2 receptor production in mononuclear cell cultures. The lack of impairment or, perhaps, even a beneficial influence on the immunodefense system may be an important property of azithromycin, especially in immunocompromised individuals. PMID- 8239586 TI - Effects of diltiazem on netilmicin-induced nephrotoxicity in rabbits. AB - Aminoglycoside nephrotoxicity remains a common clinical problem and is the major cause of acute toxic renal failure in hospitalized patients. In recent studies, calcium channel blockers gave controversial results in the prevention of acute ischemic or toxic renal failure. The aims of the study were (i) to describe a rabbit model of mild renal failure (50% reduction in glomerular filtration rate with a mean value of 1.78 +/- 0.46 ml/kg/min) induced by netilmicin given intramuscularly at 20 mg/kg of body weight every 8 h for 5 days, (ii) to investigate the protective effect of diltiazem given at a therapeutic dose (1 mg/kg given intramuscularly every 8 h for 5 days), and (iii) to investigate the mechanisms of this protection through evaluation of function tests, optic histology, and glomerular morphometry. Animals treated with netilmicin and diltiazem exhibited an unchanged glomerular filtration rate compared with controls (3.39 +/- 0.58 versus 3.68 +/- 0.78 ml/kg/min, respectively). This protective effect was not associated with any change in systemic or renal hemodynamics (i.e., no change in renal plasma flow) or changes in the pharmacokinetics of netilmicin, as assessed by fractional excretion and cortical uptake. Netilmicin-induced tubular toxicity was unchanged by diltiazem. Our results suggest that (i) netilmicin exhibits a toxic effect at both the glomerular and the tubular levels, (ii) diltiazem, a calcium channel blocker, when given at low therapeutic doses, is able to prevent the aminoglycoside induced renal failure through a potential glomerular mechanism. The precise mechanisms of the protection remain to be elucidated. These results deserve clinical evaluation in high-risk patients. PMID- 8239587 TI - Anti-Mycobacterium avium activity of quinolones: in vitro activities. AB - The MICs of 88 quinolones against 14 selected reference and clinical strains of Mycobacterium avium-M. intracellular complex were determined. Agents tested included ciprofloxacin, sparfloxacin (PD 131501), and 86 other experimental quinolones. Test strains were selected to represent various susceptibilities to ciprofloxacin and other drug resistance profiles. MICs were determined by the microdilution method in 7HSF broth, with incubation for 14 days at 35 degrees C. The results showed 25 of the quinolones to be active against the strains, with MICs for 90% of the strains (MIC90s) of 2 to 32 micrograms/ml. Ten of these compounds had activities equivalent to or greater than that of ciprofloxacin. The most active compound was PD 125354, with an MIC50 of 0.5 micrograms/ml and an MIC90 of 2 micrograms/ml; comparable values for ciprofloxacin were 4 and 8 micrograms/ml, respectively. The next most active compounds, with MIC90s of 4 micrograms/ml, were sparfloxacin (PD 131501), PD 123982, PD 135144, and PD 119421. MIC90s of PD 131575, PD 126889, PD 122642, PD 139586, and PD 143289 were 8 micrograms/ml. Further evaluation of the most active agents is warranted, as is assessment of structure-activity relationships of active and inactive agents to elucidate the active portions of the compounds and to lead to the development of compounds with enhanced activity. PMID- 8239588 TI - Anti-Mycobacterium avium activity of quinolones: structure-activity relationship studies. AB - The structures and inhibitory activities of 88 quinolones, previously studied as potential in vitro inhibitors of 14 selected strains of Mycobacterium avium complex, were examined in an effort to identify a quinolone with optimal activity towards all strains. A MULTICASE structure-activity relationship analysis of the inhibitory activities of these 88 quinolones against 14 strains of M. avium was performed and led to the identification of a number of structural constraints required to overcome the resistance of most of the strains. Our data suggested that the increased resistance of the strains was probably not due to a specific resistance mechanism but rather due to gradual limitation of the constraints imposed on the structure of the quinolones. This increasing structural selectivity could be produced either at the level of cell membrane penetration or at the level of interaction with the DNA gyrase receptor site. On the basis of these findings, a number of new quinolones holding the promise of superior activity are currently being evaluated in vitro and in vivo to determine the clinical relevance of our observations. PMID- 8239589 TI - In vitro and in vivo disposition and metabolism of 3'-deoxy-2',3' didehydrothymidine. AB - The disposition and metabolic fate of 3'-deoxy-2',3'-didehydrothymidine (D4T) were evaluated both in isolated hepatocytes and in nonhuman primates. Rapid formation of thymine and beta-aminoisobutyric acid (BAIBA) occurred following incubation of hepatocytes with 10 microM [5(-3)H]D4T. Substantial levels of tritiated water were also detected. Exposure of cells to D4T in the presence of either 1 mM thymine or 10 microM benzyloxybenzyluracil, an inhibitor of dihydropyrimidine dehydrogenase, decreased intracellular BAIBA levels by approximately 89 and 63%, respectively. Concurrently, [3H]thymine levels increased two- to fivefold. These results are consistent with D4T being cleaved to thymine, which is then degraded to BAIBA. A similar metabolic disposition was observed in monkeys following administration of 25 mg of [5(-3)H]D4T per kg of body weight. BAIBA, thymine, and tritiated water were identified in plasma and urine. Approximately 50% of the administered dose was recovered in urine within 24 h, with the majority of the radioactivity representing unchanged drug. After administration intravenously or orally of 25 mg of [4(-14)C]D4T per kg of body weight to monkeys, a novel metabolite, designated X, in addition to unchanged D4T, thymine, and BAIBA, was also detected. The sum of the three metabolites and unchanged drug accounted for virtually all of the radioactivity in plasma and urine. Thymine and X exhibited kinetic profiles similar to that of D4T, with plasma elimination half-life of 2 to 3 h, whereas BAIBA levels remained constant for extended periods and declined slowly; this metabolite could be detected 24 h after intravenous drug administration. Mean oral bioavailability of D4T was high at approximately 70%. As observed in the [5(-3)H]D4T study performed in monkeys, approximately half of the administered [4(-14)C]D4T was recovered unchanged. The remainder was not recovered in urine or feces collected up to 30 days after drug administration. These data suggest that D4T metabolites are further metabolized by salvage pathways and/or converted to biological macromolecules. PMID- 8239590 TI - In vitro and in vivo activities of Q-35, a new fluoroquinolone, against Mycoplasma pneumoniae. AB - The in vitro potency and in vivo efficacy of Q-35, a new fluoroquinolone, against Mycoplasma pneumoniae were investigated by pharmacokinetic studies with M. pneumoniae-infected hamsters. By using fluoroquinolones, macrolides, and tetracyclines as references, Q-35 was found to possess the greatest mycoplasmacidal activity. The MIC for 90% of strains tested (MIC90) and the MIC50 were 0.78 and 0.39 microgram/ml, respectively, and the MBC for 90% of strains tested (MBC90) and the MBC50 were 3.13 and 0.78 microgram/ml, respectively. The MBC50-to-MIC50 ratio for Q-35 was 2. Furthermore, only Q-35 continued to be effective against 19 strains of erythromycin-resistant mutants of M. pneumoniae. The efficacies of fluoroquinolones against M. pneumoniae were also investigated by using an experimental hamster pneumonia model to measure the CFU of M. pneumoniae in the lungs. Q-35 and ofloxacin were efficacious following oral administration of 200 mg/kg/day for 5 days, initiated 24 h after infection, while ciprofloxacin was not active. Continuous administration of Q-35 for 10 days significantly reduced numbers of viable M. pneumoniae in the lungs. These results suggest that both Q-35 and ofloxacin are effective in the early phase of infection and, moreover, that Q-35 is also effective in the middle stage of infection, when progressive lung alterations and continuous increases in mycoplasmal growth occur. Peak levels of Q-35 in sera and lungs after oral administration were higher than those of ciprofloxacin but lower than those of ofloxacin. On the basis of these results, Q-35 appears to be a promising antimicrobial agent in chemotherapy of mycoplasmal infection. PMID- 8239591 TI - Quinolones in treatment of human brucellosis: comparative trial of ofloxacin rifampin versus doxycycline-rifampin. AB - Quinolones have been reported to be active against Brucella species in vitro. In this prospective randomized study, the efficacy and safety of the combination of ofloxacin plus rifampin were compared with the efficacy and safety of doxycycline plus rifampin, both combinations administered for a 6-week period in treatment of brucellosis. Sixty-one patients were enrolled in the study, and 49 had blood or bone marrow cultures positive for Brucella melitensis. Thirty patients received 200 mg of doxycycline plus 600 mg of rifampin once daily, and 31 patients were treated with 400 mg of ofloxacin plus 600 mg of rifampin once daily for 6 weeks. Nine patients in each group had complications of the disease. There was one therapeutic failure in the ofloxacin-rifampin treatment group, and one patient from each group relapsed (3.3% of those in the doxycycline-rifampin treatment group versus 3.2% of those in the ofloxacin-rifampin treatment group). Gastric discomfort was the major side effect observed in 13 patients (43.3%) who received doxycycline plus rifampin, whereas only 2 patients (6.5%) treated with ofloxacin plus rifampin complained of gastric irritation. These results suggest that the combination of ofloxacin plus rifampin administered for 6 weeks is as effective as doxycycline plus rifampin given for the same period, regardless of the presence of complications of the disease. PMID- 8239592 TI - Comparative pharmacokinetics and serum bactericidal activities of SCE-2787 and ceftazidime. AB - Ceftazidime and the new SCE-2787 are parenteral cephalosporins with a broad antimicrobial spectrum. Pharmacokinetics, serum bactericidal activities, and side effects were investigated in a randomized crossover study. A total of 12 healthy volunteers received a 20-min infusion of 1.5 g of SCE-2787 or 2.0 g of ceftazidime. Serum and urine concentrations were determined by the bioassay method and by high-pressure liquid chromatography (HPLC). The mean (+/- standard deviation) drug concentrations in serum at the end of infusion of SCE-2787 and ceftazidime were 124.4 +/- 23.8 and 233.1 +/- 54.1 mg/liter, respectively. The urine recovery of SCE-2787 was 87.8% +/- 5.5% of dose in 24 h and for ceftazidime was 85.8% +/- 6.3% of dose in 24 h. Metabolites of SCE-2787 could not be detected by HPLC in serum or urine. Pharmacokinetic parameters were calculated both with a noncompartmental analysis and on the basis of an open two-compartment model (drugs are administered into and eliminated from a central compartment only. However, reversible drug distribution from the central space occurs simultaneously into one peripheral space). The area under the concentration time curve from 0 h to infinity of SCE-2787 was 197.9 +/- 25.4 mg.h/liter, and that of ceftazidime was 334.2 +/- 40.0 mg.h/liter. SCE-2787 had a mean terminal half-life in the elimination phase of 109.0 +/- 15.3 min, while that of ceftazidime was 99.0 +/- 13.4 min. The volume of distribution at steady state of SCE-2787 was 17.1 +/- 1.6 liters/70 kg, and that of ceftazidime was 122.9 +/- 1.3 liters/70 kg. The mean residence time of SCE-2787 was 136.4 +/- 15.4 min, and that of ceftazidime was 122.9 +/- 12.7 min. The renal clearance per. 1.73 m2 of SCE-2787 was 103.1 +/- 12.3 ml/min, and that of ceftazidime was 80.6 +/- 13.2 ml/min. The serum bactericidal activities were measured with the microdilution method of Stratton and Reller (L. B. Reller and C. W. Stratton, J. Infect. Dis. 136:196 204, 1977) against 40 clinically isolated strains. One hour after administration, we measured mean reciprocal bactericidal titers of SCE-2787 and ceftazidime, respectively, against Escherichia coli of 388 and 243, against Klebsiella pneumoniae of 395 and 138, against Pseudomonas aeruginosa of 13.0 and 12.7, and against Staphylococcus aureus of 32.2 and 4.0. No severe side effects were observed in this single drug administration. PMID- 8239593 TI - Antileishmanial activity of sodium stibogluconate fractions. AB - Sodium stibogluconate, a pentavalent antimony derivative produced by the reaction of stibonic and gluconic acids, is the drug of choice for the treatment of leishmaniasis. It has been reported to be a complex mixture rather than a single compound. We separated sodium stibogluconate into 12 fractions by anion-exchange chromatography. One fraction accounted for virtually all the leishmanicidal activity of the fractionated material against Leishmania panamensis promastigotes, with a 50% inhibitory concentration (IC50) of 12 micrograms of Sb per ml; that of unfractionated sodium stibogluconate was 154 micrograms of Sb per ml. Further analysis of this active fraction revealed that a major component was m-chlorocresol, which had been included in the sodium stibogluconate formulation as a preservative. The IC50 of pure m-chlorocresol was 1.6 micrograms/ml, a concentration equivalent to that present in unfractionated sodium stibogluconate at a concentration of 160 micrograms of Sb per ml. After ether extraction to remove m-chlorocresol, the IC50 of sodium stibogluconate was > 4,000 micrograms of Sb per ml. In contrast, when L. panamensis amastigotes were grown in macrophages, the IC50 of ether-extracted sodium stibogluconate was 10.3 micrograms of Sb per ml. The 12 fractions of ether-extracted sodium stibogluconate obtained by anion-exchange chromatography had IC50s of 10.1 to 15.4 micrograms of Sb per ml. We conclude that preservative-free sodium stibogluconate has little activity against L. panamensis promastigotes but is highly active against L. panamensis amastigotes in macrophages. This activity is associated with multiple chemical species. PMID- 8239594 TI - Association of Torulopsis glabrata infections with fluconazole prophylaxis in neutropenic bone marrow transplant patients. AB - Because the use of fluconazole prophylaxis had been associated with an increased rate of Candida krusei infections at The John Hopkins Oncology Center, early empiric amphotericin B plus flucytosine were given to febrile neutropenic patients colonized by C. krusei. By this practice, the proportion of fungemias attributable to C. krusei was low (12.5%) in patients receiving fluconazole over a 6-month interval. However, Torulopsis (Candida) glabrata assumed a much higher proportion of fungemias (75%) among patients receiving fluconazole. In vitro susceptibility testing combined with this clinical experience suggests that some T. glabrata isolates are not susceptible to fluconazole and can cause breakthrough infections in patients receiving fluconazole. PMID- 8239595 TI - Partial characterization of Nocardia farcinica beta-lactamases. AB - The beta-lactamases obtained from culture supernatants and cell extracts of 26 clinical strains and 5 reference strains of Nocardia farcinica were partially characterized. The enzymes exhibited two patterns on isoelectric focusing (IEF). beta-Lactamases from the majority of the 31 strains (87%) including the 5 reference strains exhibited two major bands with pIs of 4.56 and 4.49. The remaining strains had two similar major bands but with slightly higher pIs. Culture supernatants and cell extracts exhibited identical patterns. The two sets of enzymes were functionally indistinguishable by substrate and inhibitor profiles and lack of inducibility. By disk testing, ampicillin, amoxicillin, ticarcillin, amoxicillin-clavulanic acid, and imipenem were highly synergistic with cefotaxime. The enzymes were primarily penicillinases and hydrolyzed cephalosporins at rates of < or = 12% of those for penicillins. N. farcinica beta lactamases were susceptible to inhibition by clavulanic acid and BRL 42715, exhibiting 50% inhibitory concentrations of 0.025 to 0.045 micrograms/ml (0.12 to 0.22 microM) and 0.05 to 0.1 micrograms/ml (0.31 to 0.63 microM), respectively, less susceptible to tazobactam, and least susceptible to sulbactam, cloxacillin, and imipenem. The beta-lactamases of N. farcinica are believed to mediate penicillin resistance and may play a secondary role in extended-spectrum cephalosporin resistance. The close similarity among N. farcinica beta-lactamases and their distinct differences from beta-lactamases of other Nocardia species support the taxonomic identity of this species. PMID- 8239596 TI - Age and therapeutic outcome of experimental Pseudomonas aeruginosa keratitis treated with ciprofloxacin, prednisolone, and flurbiprofen. AB - This study was conducted to determine whether the age of the host influences the pathogenesis and therapeutic outcome of drug-treated Pseudomonas aeruginosa keratitis. Young (3- to 5-month-old) and old (1.5- to 3-year-old) rabbits were intrastromally infected with P. aeruginosa ATCC 27853. Sixteen hours later, rabbits in both age subpopulations were divided into three groups and treated topically as follows: group 1, phosphate-buffered saline; group 2, 0.3% ciprofloxacin; and group 3, 0.3% ciprofloxacin, 1.0% prednisolone, and 0.03% flurbiprofen. Drops were given every 15 min for 1 h and then every 30 min for 9 h. At 27 h postinfection, ocular pathology was graded with a slit lamp examination (SLE) scoring system. Aqueous humor was collected for ciprofloxacin quantitation, and corneas were harvested for bacterial enumeration and estimation of polymorphonuclear leukocytes. Young rabbits had more severe inflammation and pathology than old rabbits. At 27 h postinfection, SLE scores and polymorphonuclear leukocyte numbers were significantly higher for young rabbits than old rabbits (P < 0.02), regardless of treatment. Prednisolone and flurbiprofen significantly reduced SLE scores in both age groups (P < 0.03) without affecting the antimicrobial efficacy of ciprofloxacin. PMID- 8239597 TI - Ex vivo pharmacodynamic study of piperacillin alone and in combination with tazobactam, compared with ticarcillin plus clavulanic acid. AB - Ten volunteers received piperacillin (4 g), piperacillin (4 g) plus tazobactam (0.5 g) (Tazocin), and ticarcillin (3 g) plus clavulanic acid (0.2 g) (Timentin) intravenously over 30 min in a cross-over blinded scheme. Blood samples were obtained 0.5 and 3 h after the end of infusion to measure by (high-pressure liquid chromatography) the concentration and bactericidal titers against 70 gram negative bacilli. Serum time-kill curves were done against 35 strains to measure killing rates and area under the time-kill curve. Using the measure of serum bactericidal activity, ticarcillin-clavulanic acid and piperacillin-tazobactam were equally effective against Pseudomonas aeruginosa, Escherichia coli, Enterobacter cloacae, Serratia marcescens, and Bacteroides fragilis. Piperacillin tazobactam was superior to ticarcillin-clavulanic acid against piperacillin resistant Klebsiella pneumoniae (4 to 16 times) and S. marcescens (2 to 4 times). By using the area under the time-kill curve, piperacillin-tazobactam was equivalent to ticarcillin-clavulanic acid against piperacillin-susceptible strains; piperacillin-tazobactam was significantly more active than piperacillin against piperacillin-resistant strains and was more active than ticarcillin clavulanic acid when the sample obtained 3 h after the end of infusion to volunteers was considered. Serum piperacillin concentrations (mean +/- standard error of the mean; in mg/liter) were 115 +/- 13 at 0.5 h and 7.4 +/- 1.4 at 3 h after the administration of piperacillin alone and 105.5 +/- 12.6 (0.5 h) and 7.7 +/- 1.6 after the administration of piperacillin-tazobactam. Serum tazobactam concentrations (in milligram per liter) were 13.1 +/- 1.4 at 0.5 h and 1.2 +/- 0.2 at 3 h. The piperacillin-tazobactam ratio was 8 +/- 0.3 at 0.5 h and 6.2 +/- 0.5 at 3 h. Piperacillin-tazobactam appears promising against beta-lactamase producing gram-negative bacilli. PMID- 8239598 TI - Liposome-encapsulated gentamicin treatment of Mycobacterium avium-Mycobacterium intracellulare complex bacteremia in AIDS patients. AB - TLC G-65, a liposome-encapsulated gentamicin, was given intravenously twice weekly for 4 weeks to AIDS patients with Mycobacterium avium-M. intracellulare complex (MAC) bacteremia at 1.7 mg of gentamicin per kg of body weight per infusion (4 patients), 3.4 mg/kg (10 patients), and 5.1 mg/kg (7 patients). MAC colony counts in blood fell by 75% or more in all three groups (P < 0.005). Drug resistance did not emerge during the study period. Transient renal insufficiency developed in one patient; no other adverse effects were detected. Liposome encapsulated gentamicin is a potential therapy for MAC infections in AIDS patients. PMID- 8239599 TI - Concentrations in plasma and tissue penetration of ceftriaxone and ornidazole during liver transplantation. AB - Plasma and epiploic fat drug concentrations and fat penetration of ceftriaxone and ornidazole given for antimicrobial prophylaxis were studied in 11 patients scheduled for liver transplantation. Ceftriaxone (1 g) and ornidazole (500 mg) were infused during 30 min after the induction of anesthesia. Arterial blood and epiploic fat samples were collected at 30, 60, and 120 min and then every 90 min following the end of the infusion until closure of the peritoneum. Blood samples were immediately centrifuged, and plasma and fat were stored at -35 degrees C until analysis. Ceftriaxone and ornidazole concentrations were determined by high performance liquid chromatography. Surgery lasted 440 +/- 84 min and required a mean of 9.5 units of packed erythrocytes and 13 units of fresh frozen plasma. Plasma ceftriaxone concentrations decreased from 89 +/- 34 to 41 +/- 16.5 micrograms/ml from the beginning of the operation until the time of closure of the peritoneum. Corresponding levels in epiploic fat decreased from 8.7 +/- 3.3 to 4.5 +/- 3.5 micrograms/g. Ornidazole concentrations ranged, respectively, between 8.7 +/- 2.5 and 4.9 +/- 1.7 micrograms/ml in plasma samples and 4.6 +/- 1.2 and 2.5 +/- 1.1. micrograms/g in fat samples. Rates of penetration into the omentum remained at about 9% for ceftriaxone and between 50 and 70% for ornidazole. Tissue ceftriaxone concentrations were, in all cases, greater than typical MICs for 90% for Escherichia coli and Klebsiella isolates tested (MIC90S). They were insufficient in 40% of patients after 60 min with regard to the MIC90S for Staphylococcus aureus. Tissue ornidazole concentrations were not superior to MIC90S for anaerobes after 30 min in 50% of patients. These results show that a single dose of 1 g of ceftriaxone provides adequate coverage against gram-negative bacteria and that 1 g instead of 500 mg ornidazole may provide a protective effect against anaerobes during liver transplantation. Prophylaxis against S. aureus and Streptococcus faecalis requires more specific antibiotics. Prophylaxis for patients with significant blood loss or initial severe renal or hepatic failure needs further evaluation. PMID- 8239600 TI - Accumulation of a newly developed fluoroquinolone, OPC-17116, by human polymorphonuclear leukocytes. AB - The accumulation of OPC-17116 by human polymorphonuclear leukocytes was studied by comparison with three other new quinolones, ofloxacin, levofloxacin, and DR 3354. The intracellular/extracellular concentration ratio was the highest for OPC 17116, being 66.2, followed by 9.8 for levofloxacin, 7.6 for ofloxacin, and 5.8 for DR-3354. Furthermore, it was suggested that the accumulation of these new quinolones was partially related to their active transport system. The elution of OPC-17116 and ofloxacin from cells was rapid, but the elution of OPC-17116 decreased at a high extracellular pH and increased at a low extracellular pH. The accumulation of OPC-17116 as well as that of the other new quinolones was satisfactorily high, thus indicating that they are expected to be useful for the treatment of various kinds of infections, particularly infections caused by cytozoic bacteria. PMID- 8239601 TI - Adverse effects of pefloxacin in irradiated C3H/HeN mice: correction with glucan therapy. AB - Opportunistic bacterial infections are the predominant cause of death following myelosuppressive radiation exposure. When used alone, a variety of immunomodulators and antibiotics have been reported to reduce radiation-induced death. In these studies, the combined therapeutic effects of the immunomodulator glucan and the quinolone antibiotic pefloxacin were evaluated for survival enhancing effects in myelosuppressed C3H/HeN mice. Mice were exposed to 7.9 Gy of whole-body 60Co radiation and treated with saline, glucan (250 mg/kg of body weight intravenously, 1 h after irradiation), pefloxacin (64 mg/kg/day orally, days 3 to 24 after irradiation), or glucan plus pefloxacin. Survival 30 days after irradiation in mice receiving these respective treatments was 25, 48, 7, and 85%. Evaluation of granulocyte-macrophage progenitor cell (GM-CFC) recovery in mice receiving these treatments revealed that, compared with recovery in saline-treated mice, glucan stimulated GM-CFC recovery, pefloxacin suppressed GM CFC recovery, and glucan administered in combination with pefloxacin could override pefloxacin's hemopoietic suppressive effect. PMID- 8239602 TI - Attachment of an aminoglycoside, amikacin, to implantable collagen for local delivery in wounds. AB - Cultured skin substitutes consisting of implantable collagen (COL) and cultured human skin cells often fail clinically from destruction by microbial contamination. Hypothetically, addition of selected antimicrobial drugs to the implant may control microbial contamination and increase healing of skin wounds with these materials. As a model for drug delivery, bovine skin COL (1 mg/ml) and amikacin (AM; 46 micrograms/ml) were modified by covalent addition of biotin (B COL and B-AM, respectively) from B-N-hydroxysuccinimide and bound together noncovalently with avidin (A). B-COL was incubated with A and then with B peroxidase (B-P) or by serial incubation with B-AM and B-P, before P-dependent chromogen formation. Colorimetric data (n = 12 per condition) from spot tests on nitrocellulose paper were collected by transmission spectrophotometry. Specificity of drug binding in spot tests was determined by (i) serial dilution of B-COL; (ii) reactions with COL, AM, or P that had no B; (iii) removal of A; or (iv) preincubation of B-COL-A with B before incubation with B-P. Binding of B-AM was (i) dependent on the concentration of B-COL; (ii) specific to B-COL, A, and B P (P < 0.05); and (iii) not eluted by incubation in 0.15 or 1.0 M NaCl. B-AM was found to block binding of B-P to the B-COL-A complex and to retain bacteriocidal activity against 10 clinical isolates of wound bacteria in the wet disc assay. Antimicrobial activity of B-AM was removed from solution by treatment with magnetic A and a permanent magnet. These results suggest that selected antimicrobial drugs can be biotinylated for attachments to COL-cultured cell implants without loss of pharmacologic activity. Because this chemistry utilizes a common ligand, any molar ratio of agents may be administered simultaneously and localized to the site of implantation. PMID- 8239603 TI - Characterization of the chromosomal aac(6')-Ii gene specific for Enterococcus faecium. AB - Chromosomal gene aac(6')-Ii of Enterococcus faecium CIP 54-32, encoding a 6'-N aminoglycoside acetyltransferase was characterized. The gene was identified as a coding sequence of 549 bp corresponding to a protein with a calculated mass of 20,666 Da. Analysis of the sequence of the deduced protein suggested that it was the second member of a subfamily of AAC(6')-I enzymes. Insertional inactivation of aac(6')-Ii led to aminoglycoside susceptibility of CIP 54-32, suggesting that this gene plays a role in resistance to AAC(6')-I substrates. The gene was detected by DNA hybridization in all 26 strains of E. faecium tested but not in 44 other enterococci of 13 species. These data suggest that the aac(6')-Ii gene is species specific and may be used to identify E. faecium. PMID- 8239604 TI - Novel antibiotic regimens against Enterococcus faecium resistant to ampicillin, vancomycin, and gentamicin. AB - Enterococci have emerged as significant nosocomial pathogens. Enterococci with resistance to commonly used antibiotics are appearing more frequently. We encountered at our institution several infections caused by Enterococcus faecium with high-level resistance to ampicillin, vancomycin, and gentamicin. The optimal antibiotic therapy for serious infections with unusually resistant enterococci has not been established. Using time-kill studies, we tested the effectiveness of various antibiotic combinations against 15 isolates of multidrug-resistant enterococci. No antibiotic was consistently effective when used alone. The combination of ampicillin plus ciprofloxacin was bactericidal for the 12 isolates for which the ciprofloxacin MIC was < or = 8 micrograms/ml. The combination of ciprofloxacin plus novobiocin also demonstrated activity against these isolates. No combination was found to be bactericidal for the remaining three isolates, which were highly ciprofloxacin resistant. These antibiotic combinations may be important for the future treatment of serious infections caused by these resistant pathogens. PMID- 8239605 TI - Identification of highly potent and selective inhibitors of Toxoplasma gondii dihydrofolate reductase. AB - Toxoplasma gondii RH was obtained in high yield from culture in RPMI medium on a line of Chinese hamster ovary cells lacking dihydrofolate reductase activity (ATCC 3952 dhfr-; American Type Culture Collection). Dihydrofolate reductase preparations from harvested organisms had specific activities of 22.9 +/- 2.1 nmol/min/mg. The 50% inhibitory concentrations against reference compounds were 0.014 microM for methotrexate, 0.24 microM for pyrimethamine, 2.7 microM for trimethoprim, and 0.010 microM for trimetrexate. The Km value for NADPH was 11 microM and followed Michaelis-Menten kinetics; the Km for dihydrofolate was ca. 11 microM, but substrate inhibition appeared to occur at high substrate concentrations. Dihydrofolate reductase from T. gondii was used to screen 130 compounds from the National Cancer Institute repository. Thirteen compounds were > 100-fold more potent than pyrimethamine toward T. gondii dihydrofolate reductase; six compounds with various potencies were 8 to 46 times as selective as pyrimethamine for the protozoal form of the enzyme over the mammalian form. Four trimetrexate analogs were more potent than trimetrexate, and two were significantly more selective. Representative compounds were also tested in a culture model of T. gondii employing uracil incorporation as an index of growth. One pyrimethamine analog was as effective as pyrimethamine in inhibiting T. gondii in culture (50% inhibitory concentration, 0.45 microM). Three other compounds were also effective at micromolar concentrations. PMID- 8239606 TI - Evaluation of a novel fluorescence polarization immunoassay for teicoplanin. AB - A fluorescence polarization immunoassay (FPI) for teicoplanin that uses the TDx Instrument System (Abbott, Irving, Tex.) as an automated analyzer has been developed by Innotron of Oregon Inc. and was evaluated in patients with staphylococcal infections enrolled in a clinical trial of the antibiotic. The assay proved accurate in estimating concentrations of between 5 and 100 mg/liter. The intraassay coefficient of variation was < 7.3%, while the interassay variance was < 11.6% against three commercially prepared standards at known concentrations of approximately 5, 35, and 75 mg/liter. Against routinely prepared standards at 10 concentrations between 5 and 100 mg/liter analyzed in a single run, the coefficient of variance did not exceed 4.3%. Compared with bioassay, the FPI demonstrated good correlation in terms of reliability (r = 0.909) in samples containing teicoplanin only and specificity (r = 0.916) in samples containing both teicoplanin and gentamicin. With a turnaround time of 20 min and with only 50 microliters of serum needed for estimation of the amount of drug in a sample, the FPI described here should provide a useful method of teicoplanin measurement in routine diagnostic laboratories. PMID- 8239607 TI - Therapy with cefoperazone plus sulbactam against disseminated infection due to cefoperazone-resistant Pseudomonas aeruginosa and Escherichia coli in granulocytopenic mice. AB - Using a granulocytopenic murine model, we evaluated the efficacy of cefoperazone plus sulbactam against disseminated infection due to isolates of beta-lactamase producing, cefoperazone-resistant (MIC, > or = 50 micrograms/ml) Escherichia coli and Pseudomonas aeruginosa. Both isolates were susceptible in vitro to cefoperazone plus sulbactam (MIC, < or = 6.3 micrograms/ml). Mice rendered granulocytopenic with cyclophosphamide were divided into three groups: group A- infected, untreated mice (controls); group B--infected, cefoperazone-treated mice (700 mg/kg of body weight); and group C--infected, cefoperazone-plus-sulbactam treated mice (700 mg plus 350 mg). In the E. coli experiment, survival rates in groups A, B, and C were 25, 46, and 73%, respectively. In the experiment with P. aeruginosa, survival rates in groups A, B, and C were 0, 10, and 50%, respectively (P < 0.001). Highly significant differences also were noted for colony counts in the blood, liver, and spleen of group C mice versus group A or B mice in both experiments. Thus, cefoperazone plus sulbactam appears to be a promising combination for the treatment of infections due to certain cefoperazone resistant gram-negative bacilli, including P. aeruginosa. PMID- 8239608 TI - Pharmacodynamic effects of extended dosing intervals of imipenem alone and in combination with amikacin against Pseudomonas aeruginosa in an in vitro model. AB - The pharmacodynamic effects of extended imipenem dosing intervals were studied against two strains of Pseudomonas aeruginosa (ATCC 27853 and an imipenem resistant mutant, 27853R) in an in vitro model of infection. Imipenem was administered as monotherapy (simulated 1-g bolus every 8 or every 12 h) and in combination with amikacin (7.5-mg/kg bolus every 12 h or a 15-mg/kg bolus once). Monotherapy with imipenem administered every 8 h was equally bactericidal at 24 h compared with regimens combined with amikacin for ATCC 27853. Imipenem administered every 12 h against the sensitive strain and both imipenem monotherapy regimens against the resistant strain demonstrated regrowth at 24 h. Although both amikacin regimens administered as monotherapy resulted in rapid bacterial killing activity with respect to time to a 99.9% reduction in log10 CFU/milliliter, regrowth at 24 h was observed at levels reaching or exceeding the initial inoculum. All combination regimens resulted in no detectable growth by 24 h regardless of dosing interval for either drug or initial susceptibility to imipenem. Results from this study indicate the potential for several novel dosing regimens against P. aeruginosa. Monotherapy with imipenem, 1 g every 8 h, was effective against a sensitive strain of P. aeruginosa. Combination therapy with imipenem and once-daily or twice-daily amikacin resulted in increased killing activity against imipenem-resistant P. aeruginosa. Once-daily or twice-daily amikacin in combination therapy, regardless of P. aeruginosa susceptibility, allowed for extension of imipenem dosing intervals. PMID- 8239609 TI - Genetic diversity of penicillin-binding protein 2B and 2X genes from Streptococcus pneumoniae in South Africa. AB - Streptococcus pneumoniae (the pneumococcus) is believed to have developed resistance to penicillin by the production of altered forms of penicillin-binding proteins (PBPs) that have decreased affinity for penicillin. Sixty-eight clinical isolates of serogroup 6 and 19 pneumococci (MICs, < 0.015 to 8 micrograms/ml) were randomly selected from hospitals across South Africa which are at substantial geographic distance from each other. The polymerase chain reaction was used to isolate the penicillin-binding domain of PBPs 2B and 2X from the chromosomal DNAs of the bacteria; the purified PBP DNA was digested with restriction enzymes, the fragments were end-labelled and separated on polyacrylamide gels, and the DNA fingerprints were visualized following autoradiography. Fingerprint analysis revealed that at least 19 PBP 2B gene variants occur in the serogroup 6 and 19 pneumococci. The PBP 2B gene revealed a uniform profile among penicillin-susceptible isolates, with variation from this profile occurring only in isolates for which MICs were > or = 0.06 micrograms/ml. Analysis of the PBP 2X gene revealed a greater diversity in the population with 26 variant genes, including some diversity among susceptible isolates. Discrete profiles of both genes were found only within narrow bands of the penicillin MIC, so that the gene pattern predicted the MIC. PBP 2X gene variation and the lack of variability among PBP 2B genes in pneumococci inhibited at low MICs confirm that PBP 2X alteration may be responsible for low-level penicillin resistance, while alterations in both PBP 2B and PBP 2X are required for high-level resistance. The extensive diversity of PBP genes in South African serogroup 6 and 19 strains suggests that altered PBP genes have arisen frequently in this population. PMID- 8239610 TI - Antiviral activity of a phosphorothioate oligonucleotide complementary to RNA of the human cytomegalovirus major immediate-early region. AB - Phosphorothioate oligonucleotides complementary to mRNA of the human cytomegalovirus (HCMV) DNA polymerase gene or to RNA transcripts of the major immediate-early regions 1 and 2 (IE1 and IE2) of HCMV were evaluated for antiviral activity in a 96-well immunoassay with primary human dermal fibroblasts as host cells. Oligonucleotides complementary to RNA of the IE2 region exhibited the most potent antiviral activity. One of these oligonucleotides, ISIS 2922, was at least 30-fold more potent than the nucleoside analog, ganciclovir, with a 50% effective concentration of 0.37 microM in the 96-well immunoassay. In an infectious virus yield reduction assay, ISIS 2922 and ganciclovir reduced production of infectious virus by 2 log units at concentrations of 2.2 and 36 microM, respectively. A control oligonucleotide showed no inhibition of virus production at concentrations as high as 3 microM. ISIS 2922 reduced IE protein synthesis in HCMV-infected cells in a dose-dependent manner which correlated with antiviral activity. The antiviral activity of ISIS 2922 was not due to oligonucleotide-induced cytotoxicity since effects on cell viability or proliferation were observed only at concentrations well in excess of effective antiviral concentrations. The specificity and potency of ISIS 2922 suggest that it may be useful for the treatment of cytomegalovirus disease in humans. PMID- 8239611 TI - Efficacy of micronized halofantrine in semi-immune patients with acute uncomplicated falciparum malaria in Cameroon. AB - Fifty subjects with acute uncomplicated falciparum malaria were treated orally with a new micronized formulation of halofantrine. The dose given corresponded to one-half the normal dose for the standard formulation. Parasitemia cleared in all subjects within 78 h. There was recrudescence of falciparum malaria in seven subjects after day 14. The mean +/- standard deviation clearance times of parasitemia and fever were 49.0 +/- 14.2 and 24.3 +/- 13.2 h, respectively. Other clinical symptoms related to malaria cleared within the first 3 days. Pruritus occurred in two subjects, back pain occurred in one subject, and diarrhea occurred in one subject; all of these symptoms were mild. Hematological and biochemical indices were not adversely affected by treatment except in five subjects in whom minor and transitory increases in aspartate aminotransferase and alanine aminotransferase were observed. Micronized halofantrine appears to be a safe, well-tolerated, and effective treatment for acute falciparum malaria in semiimmune patients. PMID- 8239612 TI - Preincubation of Candida albicans strains with amphotericin B reduces tumor necrosis factor alpha and interleukin-6 release by human monocytes. AB - The release of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) by human monocytes stimulated with whole heat-killed Candida albicans CA3 (a clinical isolate) and CA2 (a germ tube-negative mutant) either treated or not treated with amphotericin B was investigated. The optimal release of the cytokines was observed at 24 h of incubation of the yeasts with the monocytes for both TNF-alpha and IL-6. The levels ranged from 10,500 to 19,000 U/ml for TNF alpha and from 350 to 460 pg/ml for IL-6. Germ tube-negative mutant CA2 induced the release of TNF-alpha at levels significantly (P < 0.05) lower than those induced by clinical isolate CA3, while no major differences were observed between the two strains with regard to their capacity to induce the release of IL-6. In all instances, preincubation of the yeasts with a sublethal concentration of amphotericin significantly reduced cytokine production. These results suggest that drug-induced alterations of fungal outer structures may affect the interactions between the yeasts and the monocytes, resulting in a reduced level of secretion of cytokines. PMID- 8239613 TI - Fluconazole resistance in Candida glabrata. AB - We report a case of infection with Candida glabrata in which the organism became resistant to fluconazole and in which pre- and posttreatment isolates were available for comparison. The organism was cross-resistant to ketoconazole and itraconazole, in common with other azole-resistant yeasts. Fluconazole was a potent inhibitor of cytochrome P-450-dependent 14 alpha-sterol demethylase (P 450DM) in lysates of cells from both susceptible and resistant cultures (50% inhibitory concentration, 0.2 microM), indicating that resistance was unrelated to changes in P-450DM. Instead, it appeared to arise from a permeability barrier to fluconazole, since resistant cells were unable to take up radiolabelled drug. PMID- 8239614 TI - Genes involved in the regulation of beta-lactamase production in enterococci and staphylococci. AB - In enterococci, the structural gene for beta-lactamase (blaZ) is identical to blaZ from Staphylococcus aureus. However, in the enterococci studied to date, beta-lactamase is produced constitutively, whereas in staphylococci it is often inducible. Recent reports have revealed the presence of two adjacent genes upstream of the staphylococcal blaZ thought to be the antirepressor (blaR1) and repressor (blaI) genes. In the present study, beta-lactamase expression mutants of the staphylococcal beta-lactamase plasmid pI524 were generated by transposon mutagenesis with the transposon Tn917. Tn917 insertions upstream of blaZ in either blaR1 or blaI resulted in constitutive beta-lactamase production, indicating that the repressor function is lost with insertion of Tn917 into either gene. This finding supports the concept that the staphylococcal beta lactamase regulatory genes are encoded on a polycistronic mRNA. The corresponding region upstream of the enterococcal blaZ from Enterococcus faecalis HH22 was sequenced and compared with the staphylococcal blaR1 sequence. The two sequences were identical for 893 nucleotides, and then the sequences diverged completely. Therefore, in strain HH22, only 51% of the putative antirepressor gene is present and the repressor gene is also absent. In conclusion, constitutive beta-lactamase production in HH22 appears to be due to a lack of the regulatory genes blaR1 and blaI which regulate expression of blaZ in staphylococci. PMID- 8239615 TI - Simulated human serum profiles of one daily dose of ceftriaxone plus netilmicin in treatment of experimental streptococcal endocarditis. AB - We performed experiments in rats aimed at determining whether a combination of ceftriaxone (CRO) and netilmicin (NET), by using once-daily administration in rats, which simulated profiles of drug in human serum, was more effective than either agent alone in the treatment of endocarditis caused by viridans group streptococci. A programmable infusion pump system enabled the production of profiles of CRO in serum that simulate those found in humans after the intravenous administration of 2 g. The subcutaneous administration of 18 mg of NET per kg of body weight produced levels in the sera of rats comparable to those after the intravenous administration of a dose of 5 mg of NET per kg in humans. Rats with catheter-induced aortic vegetations were infected intravenously with two test strains, a CRO-susceptible Streptococcus sanguis strain (MICs of CRO and NET, 0.064 and 8 mg/liter, respectively) and a relatively CRO-resistant Streptococcus mitis strain (MICs of CRO and NET, 2 and 8 mg/liter, respectively). Against both strains, the combination of CRO and NET was synergistic in vitro as determined by time-kill curves. Treatment of rats was started 48 h postinfection and lasted for 3 days. CRO alone was effective against the susceptible strain (P < 0.001 compared with control animals) but was not effective against the resistant organism. A significantly enhanced antibacterial activity of the CRO NET combination in reducing the valvular bacterial counts was observed with both test strains (P < 0.001). The synergistic effect was obtained with a single daily injection of NET which provided detectable levels in serum for only 8 h, suggesting that in vivo synergism in the treatment of infections caused by viridans group streptococci can be obtained without 24 h of aminoglycoside coverage. These experimental data might provide a rationale for clinical trials of a once-a-day dosing regimen in the treatment of streptococcal but nonenterococcal endocarditis. PMID- 8239616 TI - Comparison of two mefloquine regimens for treatment of Plasmodium falciparum malaria on the northeastern Thai-Cambodian border. AB - In 1991 and 1992, a prospective randomized trial was conducted on the northern Thai-Cambodian border. That trial compared the efficacy and tolerance of two mefloquine regimens for the treatment of uncomplicated Plasmodium falciparum malaria in an area with multi-drug-resistant P. falciparum. The resolution of fever and other symptoms was faster with high-dose mefloquine (25 mg/kg of body weight [M25 group; n = 68]) than with the conventional 15-mg/kg dose (M15 group; n = 71). There were no early treatment failures (days 7 to 9) in the M25 group, but there were 5 (7%) treatment failures in the M15 group (P = 0.03). The incidences of treatment failures by day 28 were 40% with the M15 group and 11% with the M25 group (P = 0.0004). By day 42, these values had risen to 50 and 27%, respectively (P = 0.01). The risk of treatment failure was highest in children (relative risk, 2.1; 95% confidence interval, 1.3 to 3.4) and patients with posttreatment diarrhea (relative risk, 2.0; 95% confidence interval, 1.3 to 3.1). Over half of the recrudescences in the M25 group occurred between days 28 and 42, whereas 17% of the recrudescences in the M15 group occurred between days 28 and 42 (P = 0.02). Thus, the sensitivity of assessment was significantly increased with longer follow-up. Treatment failure was associated with a delayed parasite clearance and an inadequate hematological recovery. Upper gastrointestinal side effects and dizziness were significantly more common in the M25 group, but overall, the high dose was relatively well tolerated, in particular by children. An increase in the dose to 25 mg/kg can prolong the therapeutic use of mefloquine in areas with multi-drug-resistant P.falciparum malaria where high-grade resistance to mefloquine is still rare. PMID- 8239617 TI - Comparison of the in vitro activities of various parenteral and oral antimicrobial agents against endemic Haemophilus ducreyi. AB - The in vitro susceptibilities to various antimicrobial agents of 100 strains of beta-lactamase-producing Haemophilus ducreyi recently isolated from patients in New Orleans, La., were determined by an agar dilution method. All strains were highly susceptible to ceftizoxime, ceftriaxone, ceftazidime, azithromycin, erythromycin, ciprofloxacin, and sparfloxacin. beta-Lactam-beta-lactamase inhibitor combinations were less active, but all strains were susceptible to them. Doxycycline exhibited the poorest activity, and the rate of resistance to doxycycline varied depending on the time after inoculation that the MIC was determined. PMID- 8239618 TI - Genetically diverse ceftazidime-resistant isolates from a single center: biochemical and genetic characterization of TEM-10 beta-lactamases encoded by different nucleotide sequences. AB - Ceftazidime-resistant isolates of Escherichia coli and Klebsiella pneumoniae produced a plasmid-mediated beta-lactamase with a pI of 5.6 with biochemical characteristics comparable to those of the TEM-10 beta-lactamase. Plasmids from the two strains were nonidentical. Both TEM-10 sequences differed from TEM-1 by substitutions of Ser-162 and Lys-237. The nucleotide sequences of the two genes were identical except for three silent nucleotide substitutions corresponding to the nucleotide differences in the Tn2 TEM-1 or Tn3 TEM-1 genes. The original TEM 10 plasmid was identical to that found in the E. coli isolate and coded for a gene that corresponded to the TEM-10 beta-lactamase from Tn2. PMID- 8239619 TI - In vitro evaluation of the activities of azithromycin alone and combined with pyrimethamine against Toxoplasma gondii. AB - By using an in vitro microassay to assess drug interaction, azithromycin combined to pyrimethamine was found more active than pyrimethamine alone against Toxoplasma gondii, and additivity between those drugs was demonstrated. Our results show that the combination of azithromycin and pyrimethamine may lead to a more rapid control of T. gondii. PMID- 8239620 TI - Susceptibility of Mycobacterium kansasii to ofloxacin, sparfloxacin, clarithromycin, azithromycin, and fusidic acid. AB - The MICs of ofloxacin, sparfloxacin, clarithromycin, azithromycin, and fusidic acid for clinical isolates of Mycobacterium kansasii were determined by the radiometric (BACTEC) method. All drugs except azithromycin elicited MICs for 90% of the strains tested that were lower than previously reported achievable maximum concentrations in serum. Ofloxacin, sparfloxacin, and clarithromycin had the largest maximum concentration in serum/MIC for 90% of strains ratio of the drugs tested. PMID- 8239621 TI - Detection and characterization of mupirocin resistance in Staphylococcus aureus. AB - Fourteen mupirocin-resistant Staphylococcus aureus strains were isolated over 18 months; 12 exhibited low-level resistance, while two showed high-level resistance. Highly mupirocin-resistant strains contained a large plasmid which transferred mupirocin resistance to other S. aureus strains and to Staphylococcus epidermidis. This plasmid and pAM899-1, a self-transferable gentamicin resistance plasmid, have molecular and biologic similarities. PMID- 8239622 TI - Ofloxacin susceptibilities of 5,667 Neisseria gonorrhoeae strains isolated in Hong Kong. AB - Of 5,667 strains of Neisseria gonorrhoeae isolated from the Government Social Hygiene (sexually transmitted disease) Clinics in Hong Kong from 1990 to 1992, there was a trend toward an increase in the percentage of strains resistant in vitro to 0.01 and 0.1 microgram of ofloxacin per ml, with 54.3 and 5.5% resistant strains, respectively, in January 1990, rising to 95.3 and 41.5%, respectively, in December 1992. The percentage of strains for which the MIC is > 1 microgram/ml remains stable, and no clinical failure has yet been seen. This trend of decreasing susceptibility warrants close monitoring when ofloxacin is used as first-line treatment for gonorrhea. PMID- 8239623 TI - In vitro activity of biapenem against clinical isolates of gram-positive and gram negative bacteria. AB - The in vitro activity of biapenem, a new carbapenem previously designated L-627, was compared with those of imipenem and several other antimicrobial agents against 771 clinical bacterial isolates. Against gram-positive organisms, biapenem was found to be approximately as active as imipenem, inhibiting 90% of isolates of most species at concentrations within one dilution of the MIC of imipenem for 90% of the isolates. Against gram-negative organisms and Bacteroides fragilis, biapenem was at least as active as and often more active than imipenem, with MICs for 90% of the isolates two- to eightfold lower than those of imipenem. PMID- 8239624 TI - In vitro activities of BAY Y3118, ciprofloxacin, ofloxacin, and fleroxacin against gram-positive and gram-negative pathogens from respiratory tract and soft tissue infections. AB - BAY Y3118 was highly active against Moraxella catarrhalis, Haemophilus influenzae, Legionella pneumophila, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus (except quinolone-resistant, methicillin-resistant S. aureus), Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus agalactiae, and Streptococcus pneumoniae (MIC for 90% of strains tested [MIC90], 0.063 micrograms/ml). For Enterococcus faecalis and Corynebacterium jeikeium, MIC90s were 4 and 2 micrograms/ml, respectively. BAY Y3118 was as active as ciprofloxacin against Pseudomonas aeruginosa (MIC90, 0.5 micrograms/ml) and had potent activity against Bacteroides fragilis (MIC90, 0.5 micrograms/ml). PMID- 8239625 TI - Novel, plasmid-encoded, TEM-derived extended-spectrum beta-lactamase in Klebsiella pneumoniae conferring higher resistance to aztreonam than to extended spectrum cephalosporins. AB - A clinical isolate of Klebsiella pneumoniae was more resistant to aztreonam than to cefotaxime and ceftazidime. It produced a clavulanate-susceptible beta lactamase with an isoelectric point of 6.3 which readily hydrolyzed penicillins, cefotaxime, and ceftazidime, but which hydrolyzed aztreonam poorly. The enzyme was encoded by a gene on a 15-kb plasmid; the gene hybridized with an intragenic DNA probe of blaTEM. PMID- 8239626 TI - Use of single-dose ofloxacin to eradicate tonsillopharyngeal carriage of Neisseria meningitidis. AB - After an outbreak of three cases of serogroup B meningococcal disease at a Norwegian college, 84 of 392 (21%) subjects were tonsillopharyngeal carriers of Neisseria meningitidis. To eradicate meningococcal carriage, 80 volunteers received a single dose of 400 mg of ofloxacin. Three days after treatment, all 75 evaluable volunteers were culture negative for N. meningitidis, and after 7 days none carried the strain that they carried initially, as judged by DNA fingerprinting. A single dose of ofloxacin was found to be 97.2% effective in eradicating carriage of N. meningitidis for a period of 33 days. The carriage acquisition rate among treated students was four times higher than that among nontreated noncarriers (P = 0.02). After ofloxacin treatment, no case of meningococcal disease occurred for 6 months. Ofloxacin may thus prevent the outbreak and spread of meningococcal disease. PMID- 8239627 TI - Lincosamide antibiotics stimulate dissociation of peptidyl-tRNA from ribosomes. AB - At nonpermissive temperatures the peptidyl-tRNA hydrolase of pth(Ts) bacterial mutants is inactivated, and cells accumulate peptidyl-tRNA and die. Doses of erythromycin, lincomycin, or clindamycin that inhibited the growth of antibiotic hypersensitive DB-11 pth+ cells accelerated the killing of DB-11 pth(Ts) cells at nonpermissive temperatures. Erythromycin and lincomycin also stimulated the accumulation of peptidyl-tRNA. Lincomycin and clindamycin stimulated peptidyl tRNA dissociation from ribosomes. PMID- 8239628 TI - Comparison of fluconazole and amphotericin B for treatment of experimental Candida endocarditis caused by non-C. albicans strains. AB - Amphotericin B and fluconazole were compared for the treatment of experimental Candida endocarditis caused by Candida tropicalis and C. parapsilosis. Rabbits received no therapy, amphotericin B (1 mg/kg of body weight per day intravenously), or fluconazole (100 mg/kg/day intraperitoneally) for either 11 or 21 days. Against both species, amphotericin B and fluconazole were equally effective overall; however, amphotericin B was more rapidly fungicidal than fluconazole in vivo against C. tropicalis. PMID- 8239629 TI - Activity of a nitrofurazone matrix urinary catheter against catheter-associated uropathogens. AB - Nitrofurazone-coated urinary catheter segments inhibited 51 (75%) of 70 urinary bacterial isolates from patients with indwelling catheters. Inhibition zones correlated significantly with the nitrofurazone MIC (r2 = 0.79, P = 0.0001). All strains except the Pseudomonas spp. were inhibited by < or = 64 micrograms of nitrofurazone per ml. MICs of nitrofurazone and nitrofurantoin correlated significantly (r2 = 0.93, P = 0.0001). PMID- 8239630 TI - Influence of circadian-stage-dependent dosing schedule on nephrotoxicity and pharmacokinetics of isepamicin in rats. AB - Nephrotoxicity was more marked in rats receiving isepamicin at midlight than at middark. And, the once-daily administration at middark induced a lesser degree of nephrotoxicity than the twice-daily injection, which indicates that the once daily treatment therapy may have potential value in the clinical use of aminoglycosides. PMID- 8239631 TI - Dynamic contrast: a sensory contribution to palatability. AB - By focusing on a sensory model for palatability, the "ice-cream effect", we speculate on some principles underlying palatability. We propose that the most highly palatable foods are likely to have higher levels of "dynamic contrast" (moment-to-moment sensory contrast from the everchanging properties of foods manipulated in the mouth). The sensory feature contributing most to dynamic contrast seems to be somatosensation. Texture, for example, over the course of masticating a crunchy food, changes markedly such that each alteration contributes differently and dynamically to sensory contrast. Our purpose herein is to propose that dynamic contrast contributes significantly to palatability of foods and beverages. PMID- 8239632 TI - Snacking patterns in obese French women. AB - Snacking patterns were studied in 273 obese women by dietary history. Snacking was observed in 60% of the sample and occurred mainly during the afternoon. Total daily energy intake was higher in snackers than in non-snackers because of greater consumption of food at meals and between meals. A variety of foods were consumed as snacks (mean macronutrient content of snacks: 52% carbohydrate (CHO), 37% lipid and 12% proteins). CHO-rich protein-free snacks were a minority (20%). Only 7% of subjects were CHO-rich protein-free snackers. The eating pattern described by Wurtman in "carbohydrate cravers" was reported by less than 10% of these women. We conclude that, in Paris, snacking plays an important role in increasing energy consumption in obese women but macronutrient-specific snacking is not frequent. PMID- 8239633 TI - Effects of flavor and macronutrient composition of food servings on liking, hunger and subsequent intake. AB - The effects of consuming foods with different macronutrient compositions and flavors on hedonic changes and development of satiety were investigated. Subjects rated their hunger and liking of a set of foods (rating set) before and after eating a serving (preload) of one of the foods in the rating set. The liking of the preload foods dropped more than the liking of the uneaten foods. Foods having the same flavor as the preload generally dropped more in liking than foods having similar macronutrients. The drops in liking increased with the caloric content of the preload but were unrelated to specific macronutrients. Less weight and calories of food were eaten after the high calorie preloads. Eating the high protein or the high-carbohydrate preload decreased hunger more than eating the high-fat food. Eating a high-protein preload decreased the weight of food eaten more than eating a high-fat or a high-carbohydrate preload and decreased total caloric intake more than eating a high-fat preload. However, macronutrient intake was not differentially affected by the macronutrient composition of a preload. Sensory-specific satiety appears to be more related to the sensory characteristics of a food than to the macronutrient composition of a food. PMID- 8239634 TI - Current attitudes and future influences on meat consumption in the U.K. AB - A survey of 1018 U.K. residents was conducted using the basic structure of the theory of reasoned action along with scenarios of possible future events investigating meat-eating and vegetarianism. Over a quarter (28%) of the sample considered themselves to be reducing meat consumption although a comparison of present and retrospective meat consumption indicated that less than a quarter of this group had cut down on a variety of meats in their diets over the past year. The attitudes related to present meat consumption were healthiness, taste, value for money and, to some extent, ethical issues. Healthiness, taste and concerns over additives were related to changes in meat consumption over the preceding year. Beliefs about the healthiness of meat were also related to reported changes in meat-eating behaviour when "something that could possibly happen in the future" was described. Future events found to affect people's estimated meat eating were the availability of polyunsaturated meat and meat produced with strict safety guarantees, the former change being dependent on a constant price. Reductions in the fat content of meat or claims extolling the nutritional benefits of eating meat were not related to predicted future meat-eating. PMID- 8239635 TI - Food deprivation reduces rats' oil preference. AB - Rats normally prefer dilute oil suspensions over vehicle, but this preference can be abolished by exposing them to dilute oil suspensions during a period of food deprivation. In the first series of experiments, rats were given a choice of 0.5 0.9% triolein vs. vehicle for 4 consecutive days each week for 3 weeks. Food was available ad libitum on days 1, 2 and 4; no food was available on day 3. Preference for oil suspension dropped sharply after, but not during, each occasion that food was withheld. Similar experiments using starch, Polycose, or sucrose did not show comparable deprivation-induced changes in preference. The second series of experiments included separate control and experimental groups. The experimental group was food deprived for 1 day when test fluids were available. The control group was food deprived the day after the test fluids were available. Food was returned and the test fluids were removed for a few days. Fluid preference was then retested with food available ad libitum. After this cycle of two preference tests was repeated 1-2 times, the experimental group showed substantially lower oil preference than did the control group. Similar results were obtained with 0.5% triolein, 0.5% soybean oil, and 5% soybean oil. A similar experiment in which a vanillin-citral flavor was used instead of oil did not show such an effect. However, preference for the vanillin-citral flavor was decreased in rats exposed to a mixture of this flavor and oil while they were food deprived. It is proposed that rats readily learn to associate oil flavor, but not other flavors, with the aversive effects of food deprivation. PMID- 8239636 TI - The rat's preference for sucrose, polycose and their mixtures. AB - In one-bottle tests (30 min/day) food-deprived rats consumed significantly more 2% Polycose than 2% sucrose, and yet in two-bottle choice tests they did not consistently prefer Polycose to sucrose. A previously proposed explanation for this effect is that rats habituate, or develop oral satiety, to the flavor of sucrose more rapidly than to Polycose, and sucrose satiety generalizes to Polycose. This idea was not supported by the results of sequential 15-min one bottle tests: the intake of 2% sucrose did not suppress subsequent Polycose intake. Instead, the failure of rats to consistently prefer Polycose to sucrose was attributed to the development of a drinking pattern that allowed the rats to mix the two solutions in their mouths. When the drinking spouts were nearby, the rats consumed similar amounts of sucrose and Polycose, but when the spouts were far apart, they consumed more Polycose than sucrose. That rats prefer the combined flavor of sucrose and Polycose to either flavor alone was confirmed in subsequent tests. Rats strongly preferred a 1% sucrose + 1% Polycose mixture to a plain 2% sucrose or 2% Polycose solution. They also preferred sucrose-Polycose mixtures to plain solutions at concentrations ranging up to 32%. The potency of the sucrose-Polycose mixture is presumably related to the activation of separate "sweet" and "polysaccharide" taste channels in the rat. PMID- 8239637 TI - Contrast in consummatory behavior. PMID- 8239638 TI - Postoperative narcotic analgesic administration. AB - The administration of narcotic analgesics to postoperative patients was examined with 180 uncomplicated adult appendectomy patients. Narcotic analgesic doses were transcribed from the hospital records of these patients for the entire postoperative period. Equianalgesic doses were calculated so that all medications were comparable with meperidine. Eighty-three percent (n = 150) of the patients received meperidine. Patients who received meperidine were given significantly more narcotic analgesics than those who received morphine sulfate. The amount of narcotic analgesics received by patients was significantly related to their length of hospital stay. Meperidine and Tylenol #3 (acetaminophen with codeine phosphate; McNeil Pharmaceutical, Spring House, PA) comprised the significantly associated analgesics. These findings suggest a need to reexamine the current use of meperidine in postoperative analgesia. PMID- 8239639 TI - Future images: an art intervention with suicidal adolescents. AB - In this study, the effectiveness of an art future-image intervention (AFI) designed to increase self-esteem, improve future time perspective, and decrease depression in hospitalized suicidal adolescents was tested. A pretest-posttest time series design was used with two groups, an experimental group and an attention placebo group. The experimental group, although hospitalized for a shorter length of time (p = .08), showed greater positive changes than the placebo group. Both groups improved on all measures during and after hospitalization. Experimental participant enthusiasm, shorter hospitalization, and positive comments at follow-up warrant continued testing and refinement of the AFI. PMID- 8239640 TI - Cholesterol screening of older persons: focusing on health education. AB - Public education about cholesterol screening and evaluation has been strongly recommended by the Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol (1988). For older persons, limited clinical evidence about benefits that may be derived from screening efforts poses particular challenges to education efforts by health care providers. Additionally, professional and lay information sources offer mixed messages about the purpose of cholesterol screenings. Studying a community hospital cholesterol-screening program provided a timely way to provide insight about the meaning of cholesterol evaluations for older persons. Findings from the program evaluation are discussed in light of the debate that continues about the benefits of cholesterol evaluation and treatment in older persons. PMID- 8239641 TI - Understanding older adult satisfaction with primary health care services at a nursing center. AB - The measurement of satisfaction levels is a positive evaluation of a distinct health care outcome. The purposes of this descriptive-correlational study were to describe the following: (a) the levels of satisfaction with primary health care services received at a nursing center (NC), and (b) the relationships between satisfaction levels and the demographic characteristics of the older adult NC clients. Older adult clients were very satisfied with the primary health care. Subscale analysis showed that the clients were most satisfied with the technical aspects of professional nursing. PMID- 8239642 TI - Is a stay in an intensive care unit a risk for falls? PMID- 8239643 TI - Increasing the response rates of reluctant professionals to mail surveys. PMID- 8239644 TI - Data management for surveys. PMID- 8239645 TI - The participation of reactive oxygen species and protein thiols in the mechanism of mitochondrial inner membrane permeabilization by calcium plus prooxidants. AB - We have recently shown that permeabilization of the inner mitochondrial membrane by calcium plus prooxidants is associated with oxidation of protein thiols forming cross-linked protein aggregates [Fagian, M. M., Pereira da Silva, L., Martins, I. S. and Vercesi, A. E. (1990) J. Biol. Chem. 265, 19955-19960]. In this study we show that mitochondria could regenerate and sustain a membrane potential (delta psi) comparable to the control experiment after the protein aggregates were cleaved by dithiothreitol. The addition of ethylene glycol bis(beta-aminoethyl ether) N,N'-tetraacetic acid, which removes Ca2+ but does not eliminate the protein aggregates, caused an incomplete and nonsustainable recovery of delta psi. Exogenous catalase prevented the disruption of membrane potential and decreased the production of membrane protein aggregates when mitochondria were incubated in the presence of Ca2+ alone or Ca2+ plus a prooxidant. This strongly indicates that H2O2 and possibly other H2O2-derived reactive oxygen species are involved in the mechanism of membrane protein aggregates production that may result in the process of membrane pore formation. PMID- 8239646 TI - Kynurenine 3-hydroxylase in brain: species activity differences and effect of gerbil cerebral ischemia. AB - We have developed a rapid and highly sensitive assay for brain kynurenine 3 hydroxylase activity. The present study determined some characteristics and species differences of kynurenine 3-hydroxylase activity in brain and evaluated the response of this enzyme to cerebral ischemia. The kynurenine 3-hydroxylase assay is based on the conversion of L-kynurenine to 3-hydroxykynurenine in vitro and the quantification of 3-hydroxykynurenine by high-performance liquid chromatography. Kynurenine 3-hydroxylase activity was detected in human, macaque, rat, mouse, and gerbil brain. Regional gerbil brain activities ranged from 20 to 50 nmol/g/h, while kynurenine 3-hydroxylase activities in other species were one order of magnitude lower. Kynurenine 3-hydroxylase was also detected in lung, kidney, spleen, intestine, and liver of gerbils, with activities larger than in brain. Delayed increases in the activity of kynurenine 3-hydroxylase occur in several brain regions following transient ischemia in gerbils. These changes are particularly marked in regions that showed the most extensive brain damage (hippocampus and striatum). Kynurenine 3-hydroxylase may have an important role in determining the flux of kynurenine in brain. PMID- 8239647 TI - Fibronectin fragments alter matrix protein synthesis in cartilage tissue cultured in vitro. AB - We reported earlier that fibronectin fragments (Fn-f) added to bovine articular cartilage cultured in serum-free culture causes marked protease expression with resultant proteoglycan (PG) degradation and release into the culture media. We have further characterized the effects of Fn-f by studies of the effects on proteoglycan, collagen, general protein, and DNA synthesis and reversibility of the cartilage damage. We report here that the most active Fn-f, a 29-kDa amino terminal Fn-f, when added to a 1 microM concentration, depressed PG and general protein synthesis in cartilage by over 50% within 24 h, as measured by sulfate and methionine/cysteine incorporation, respectively. This same Fn-f decreased PG synthesis throughout the full thickness cartilage section as shown by autoradiography. PG and general protein synthesis were significantly depressed within 24 h by 29-kDa Fn-f concentrations as low as 10 nM. Synthesis rates were effected by 100-fold lower Fn-f concentrations than was induction of proteinases. Removal of the 29-kDa Fn-f allowed a gain to supernormal levels of PG and protein synthesis. Cartilage damaged to the extent of removal of over 50% of the total PG did not replace PG after over 4 weeks in 10% serum-Dulbecco's modified Eagle minimum with or without added TGF-b1 and rIGF-a. These data show that while the effects of Fn-f on elevating protease expression and depressing PG synthesis are reversible, the resultant cartilage damage is apparently irreversible in vitro. Therefore, if Fn-f-mediated cartilage damage occurs as part of cartilage disease processes, the pathologic effects would be quite significant. PMID- 8239648 TI - Alterations in the carbohydrate structures of an abnormal protein from sera of patients with rheumatoid arthritis. AB - G4-11, an abnormal immunoglobulin G4 (IgG4) glycoprotein, highly purified from the sera of patients with rheumatoid arthritis and from healthy individuals, contains two asparagine-linked sugar chains in one molecule. Comparative studies of the sugar chains released by hydrazinolysis revealed that the structures of the sugar chains of rheumatoid arthritis G4-11 are quite different from those of normal individuals. Although all G4-11 contained biantennary complex-type oligosaccharides like in the case of serum IgGs, samples from patients with rheumatoid arthritis had less galactosylated forms than those from normal individuals. These results indicated that this galactose deficiency of the sugar chains occurs in the abnormal IgG4 molecule of rheumatoid arthritis patients, just as was found in whole serum IgG. PMID- 8239649 TI - Identification of Ser143 as the site of modification in the active site of histidine ammonia-lyase. AB - Histidine ammonia-lyase (histidase) from Pseudomonas putida was irreversibly inactivated by L-cysteine at pH 10.5 in the presence of oxygen. Inactivation was accompanied by the formation of a new uv-absorbing species centered around 340 nm. L-[35S]cysteine labeling experiments revealed that 4 mol of L-cysteine was bound per mole of enzyme tetramer upon complete modification. However, the radiolabel was dissociated from the protein under denaturing conditions without loss of the 340-nm absorbance. Prior inactivation of histidase by cyanide, borohydride, or bisulfite precluded the formation of the 340-nm species in subsequent L-cysteine modification experiments. This suggests a common target site for modification of histidase by all of these reagents. Based on its strong absorbance at 340 nm an octapeptide was isolated from L-cysteine-inactivated histidase following trypsin and staphylococcal V8 protease digestion. Electrospray MS/MS revealed that this peptide (Gly138-SerValGlyAlaSerGlyAsp145) contained an unidentified modification of mass 184 Da located on Ser143. This peptide and the serine residue are conserved in all histidases and phenylalanine ammonia-lyases for which the amino acid sequence is available. Ser143 represents the binding site for an electrophilic cofactor required for histidase activity. PMID- 8239650 TI - Purification of soluble murine interleukin 5 (IL-5) receptor alpha expressed in Chinese hamster ovary cells and its action as an IL-5 antagonist. AB - Recombinant soluble murine interleukin 5 receptor alpha (sIL-5R alpha) was expressed in Chinese hamster ovary cells and its biological activity was characterized. By using murine IL-5-coupled Sepharose, the single step purification method of the protein was established. The purified protein inhibited the murine IL-5-induced differentiation of BCL1 cells into immunoglobulin M-secreting cells in a dose-dependent manner. Moreover, it also inhibited murine IL-5-induced but not human IL-5-induced proliferation of TF-1 cells. These results indicated that sIL-5R alpha acted as an IL-5 antagonist and apparent species specificity of IL-5 action on murine cells was mediated, at least in part, by the alpha-chain of the IL-5 receptor. PMID- 8239651 TI - Actin stimulates plasmin generation by tissue and urokinase-type plasminogen activators. AB - The experiments reported here were carried out to define in greater detail actin's stimulation of plasmin generation by t-PA. Actin did not alter t-PA's hydrolysis of a synthetic substrate, and thus is unlikely to have a direct effect upon t-PA's proteolytic activity. When studied in a single-stage assay, actin accelerated t-PA-mediated plasmin generation from both Glu-plasminogen and Lys plasminogen, indicating the central role of ternary complex formation. Although actin does not appear to bind two-chain urokinase (tcu-PA), it stimulates tcu PA's cleavage of Glu-plasminogen. This finding suggests that actin alters the conformation of Glu-plasminogen to an open form. The failure of actin to increased plasmin generation by tcu-PA acting on Lys-plasminogen, which is in an open configuration, is consistent with this interpretation. Immunoglobin G, which shares with actin the property of binding to Glu-plasminogen after nicking by plasmin, did not stimulate tcu-PA's cleavage of Glu-plasminogen, indicating the uniqueness of actin's effects and suggesting interactions between actin and plasminogen at multiple binding sites. Unlike fibrin and heparin, whose stimulation of t-PA is related to polymer length actin is able to stimulate t-PA when presented in either a monomeric or polymeric form. Denaturation of actin by exposure to urea and guanidine increased its ability to stimulate plasmin generation by t-PA. Because actin's structure is maintained by a noncovalently bound adenine nucleotide (ATP or ADP), exposure to ATP/ADPases found in plasma and on cell membranes might also result in its denaturation. Actin treated with an enzyme functionally similar to such ecto-ATP/ADPases, potato apyrase, was more potent than native actin in stimulating plasmin generation by t-PA. The effects of apyrase were blocked by the addition of the plasma actin-binding proteins, gelsolin and the vitamin D-binding protein (DBP). Thus, denaturation of actin may occur in under physiologic conditions, with potential biological consequences. Actin thus appears to be unique with regard to its interactions with the fibrinolytic system and plasma actin-binding proteins may serve to protect the host from the effects of denatured actin. PMID- 8239652 TI - Thermal stability and activation of bovine lens multicatalytic proteinase complex (proteasome). AB - The multicatalytic proteinase complex (MPC; proteasome) can be isolated in a latent form which then can be activated for protein hydrolysis by physiological and nonphysiological treatments, including high temperature. In this study, the temperature dependency profiles for the hydrolysis of Cbz-Gly-Gly-Leu-pNA and Cbz Val-Gly-Arg-pNA by bovine lens MPC are found to be those expected for a thermostable enzyme, with single optima above 50 degrees C. In contrast, hydrolyses of Cbz-Leu-Leu-Glu-2NNp and alpha 2-crystallin, a lens structural protein, show two temperature transitions, indicating that hydrolysis of these substrates can be activated by elevated temperature. Temperature dependency profiles of peptidase activity in Tris-HCl compared to Hepes buffer suggest that Tris decreases the thermal stability of MPC. After 10 min preincubation in Tris HCl at 53 degrees C, lens MPC activities are reduced by 50-60% and loss of the major MPC band can be seen on nondenaturing gels. The presence of alpha 2 crystallin during preincubation partially prevents the loss of activity. Although alpha-crystallin has been reported to function as a molecular chaperone, similar protection by other MPC substrates suggests that alpha 2-crystallin stabilized the MPC as a substrate. Our findings indicate both activation and inactivation of the enzyme at elevated temperatures. It is proposed therefore that high temperature activates the MPC but to a more labile form which can be partially stabilized by protein substrates. PMID- 8239653 TI - Isolation and characterization of pectin methylesterase from papaya. AB - Pectin methylesterase (PME) (EC 3.1.1.11) has been purified to apparent homogeneity from ripe papaya fruits. The purification protocol consisted of ammonium sulphate precipitation (60-80%) and cation exchange chromatography in CM Sepharose CL-6B and Mono S. Papaya PME consists of two components (PME 1 and PME 2), which have been shown to be isoenzymes by Ferguson plot analysis. The molecular weight of the enzyme is 27,000 while its isoelectric point is greater than pH 9.0. The N-terminal sequences of PME 1 and PME 2 are SVVTPNAVVADDGVFXFKTG. Both PME 1 and PME 2 showed optimum activities at pH 8.0 and 35 degrees C. The average Kms of PME 1 and PME 2 are 0.0071 and 0.0166 g/liter pectin respectively, while the corresponding average Vmaxs are 741 and 800 mumol methanol/min/mg protein, respectively. Papaya pectin methylesterase is activated by cations, but the effect is more pronounced for divalent than monovalent cations. Inhibition studies showed that sucrose is a noncompetitive inhibitor while p-chloromercuribenzoic acid has no significant effect on its activity. PMID- 8239654 TI - Selective induction of alpha 1 isoform of (Na+ + K+)-ATPase by insulin/insulin like growth factor-I in cultured rat astrocytes. AB - Treatment of cultured rat astrocytes with insulin increased (Na+ + K+)-ATPase activity expressed per protein or DNA by 1.6- to 2.1-fold, but did not affect Mg(2+)-ATPase and adenylate cyclase activities. Insulin treatment increased protein and DNA contents under the conditions, while it did not cause morphological differentiation as determined by a microscopic inspection. Insulin like growth factor-I (IGF-I) had a similar effect on the enzyme activity in astrocytes: the effect of insulin was observed at supraphysiological concentrations, while that of IGF-I was observed at physiological concentrations. Insulin and IGF-I both stimulated DNA synthesis at the concentrations that caused an increase in enzyme activity. The effect was blocked by tyrosine kinase inhibitors such as genistein and herbimycin A and by cycloheximide. Western blot analysis showed that alpha 1 and alpha 2 isoforms of (Na+ + K+)-ATPase were present in cultured astrocytes and that insulin and IGF-I increased the content of the alpha 1 isoform but did not that of the alpha 2 isoform. Two components of ouabain inhibition were observed in the enzyme purified partially from cultured astrocytes, and treatment of the cells with IGF-I increased the ratio of the low affinity component of the inhibition, indicating a selective increase in the activity of the alpha 1 isoform. These results indicate that insulin increases (Na+ + K+)-ATPase activity through an activation of IGF-I receptors and the increase is due to the selective induction of the alpha 1 isoform in cultured astrocytes. PMID- 8239655 TI - pH of melanosomes of B 16 murine melanoma is acidic: its physiological importance in the regulation of melanin biosynthesis. AB - It was postulated by us that melanin biosynthesis cannot take place unless the melanosomal pH is acidic. That melanosomal pH may be acidic was suggested by other researchers as well. However, the actual pH was never determined. The current study was undertaken to estimate the intramelanosomal pH. Lightly melanized and heavily melanized melanosomes from B16 murine melanoma tissue were prepared by sucrose density gradient centrifugation and their internal pH was estimated. It was shown that quenching of fluorescence of 9-aminoacridine by melanosomes was to a large extent dependent on the pH difference between melanosomes and the medium in which they were suspended and it was used to determine the pH of lightly and heavily melanized melanosomes from B16 melanoma. The pH of lightly melanized melanosomes and heavily melanized melanosomes was found to be in the range of 4.2 to 4.6 and 3.0, respectively. These organelles were shown to hydrolyze fluorescein diacetate to fluorescein. The excitation spectrum of fluorescein generated in lightly melanized melanosomes and heavily melanized melanosomes also indicated that the pH in lightly melanized melanosomes was 4.6 and lower than 4.6 in heavily melanized melanosomes. The enzymes are active at such low pH. These organelles are also shown to contain proton translocating ATPase, which may be involved in the acidification of lightly melanized melanosomes, possibly in addition to melanin biosynthesis. These results confirm our original postulate that melanin biosynthesis that occurs in melanosomes takes place under acidic conditions. PMID- 8239656 TI - Overexpression, purification, and kinetic characterization of a carboxyl-terminal truncated yeast squalene synthetase. AB - Yeast squalene synthetase which has been truncated by 24 amino acids at the C terminus has been overexpressed in Escherichia coli and constitutes approximately 20% of the total soluble cell protein. For the first time, milligram quantities of this essential enzyme in the cholesterol biosynthetic pathway have been purified to near homogeneity by ammonium sulfate precipitation and Mono Q anion exchange chromatography so that the steady-state rate constants could be measured. A combination of 10% methanol, 10% glycerol, 30 mM octyl-beta-D glucopyranoside, 0.4% Brij-58, and 1 mM dithiothreitol in 25 mM sodium phosphate, pH 7.4, was essential for the stability and maximal enzyme activity of the near homogeneous enzyme. Kinetic analysis indicated a Km for farnesyl pyrophosphate of 2.5 microM, suggesting fairly tight binding of farnesyl pyrophosphate to truncated yeast squalene synthetase. The turnover number, kcat, for the conversion of farnesyl pyrophosphate to squalene was 0.53 s-1, and the apparent second order rate constant, kcat/Km, was 2.1 x 10(5) M-1 s-1, indicating a relatively slow conversion of farnesyl pyrophosphate to squalene and a low specificity constant for this enzyme. In addition, Km for NADPH and NADH was 0.5 and 3.6 mM, respectively. Moreover, truncated yeast squalene synthetase shows a preference for NADPH over NADH as reflected in the sevenfold higher kcat/Km value for NADPH similar to that for the native enzyme. PMID- 8239657 TI - Assembly of Atlantic cod (Gadus morhua) brain microtubules at different temperatures: dependency of microtubule-associated proteins is relative to temperature. AB - Isolated cod (Gadus morhua) brain microtubules were found to have a broad temperature interval for assembly. In contrast to mammalian microtubules they assembled even at as low temperatures as 14 degrees C. Evidence was found that temperature alters the dependency of microtubule-associated proteins (MAPs) for assembly. The assembly was MAPs-dependent at low, but not at higher temperatures. Assembly at +18 degrees C was inhibited by both NaCl and estramustine phosphate. These compounds are well known to inhibit the binding of MAPs to tubulin. At higher temperatures there was no MAPs dependency for assembly, despite that MAPs bound to the microtubules. Cow MAPs had the same effect as cod MAPs, suggesting that despite differences in MAP composition, the effect is not caused by the unusual composition of cod MAPs. The results therefore suggest that these differences in MAPs dependency are due to intrinsic properties of cod tubulin or tubulin-to-tubulin interactions. Small temperature-induced conformational changes of tubulin and a slight enrichment of acetylated and detyrosinated tubulin in microtubules assembled at +30 degrees C as compared to +15 degrees C, were observed. The ability to alter the assembly stimulating effect of MAPs may be important for the cell to regulate microtubule dynamics and stability. In addition, changes in tubulin conformation and composition of tubulin isoforms may reflect adaptations for microtubule assembly at low temperatures. PMID- 8239658 TI - Human erythrocyte casein kinase II: characterization and phosphorylation of membrane cytoskeletal proteins. AB - Casein kinase II activities were purified from human erythrocyte membrane and cytosolic fractions to apparent homogeneity. The kinases isolated from the membrane and cytosolic fractions exhibited the same subunit composition and the ability to utilize ATP and GTP as phosphoryl donors. Antibodies against the alpha and alpha' subunits of human casein kinase II cross reacted with the corresponding subunits of both erythrocyte casein kinases. Spermine, spermidine, putrescine, and polylysine stimulated to varying degrees the activities of erythrocyte casein kinase II, whereas heparin inhibited the kinase activities. Both kinases were found to catalyze the phosphorylation of several erythrocyte membrane cytoskeletal proteins, including spectrin, ankyrin, adducin, protein 4.1, and protein 4.9. Unlike casein kinase I, casein kinase II did not phosphorylate band 3 appreciably. A preliminary estimate indicates that both human erythrocyte membrane and cytosolic casein kinase II catalyze the incorporation of approximately 1.2 and 3.5 moles of phosphate into each mole of spectrin and ankyrin, respectively. An analysis of the phosphopeptide maps of ankyrin indicates that both membrane and cytosolic kinases phosphorylate the same domains within ankyrin. These data, taken together, suggest that the type II casein kinases isolated from human erythrocyte membrane and cytosol are either identical or closely related and may play a role in the regulation of cytoskeletal protein interactions. PMID- 8239659 TI - Secondary structure and thermal stability of caldesmon and its domains. AB - Muscle caldesmon is a long, thin protein molecule whose N- and C-terminal regions are separated by a central region which is not present in nonmuscle caldesmon. The three regions appear to be independent structural domains since the alpha helical content of intact muscle and liver caldesmon is a sum of the alpha helical contents of the component thrombic fragments over a broad temperature range. Based on circular dichroism spectra of liver and muscle caldesmon and its fragments, together with secondary structure prediction algorithms, it is estimated that the N-domain consists of a string of four to five short-to intermediate-length alpha-helices; the central domain contains a long continuous alpha-helical stretch; and the C-domain can be divided into two subregions, the N terminal C1-region, containing a long alpha-helix, and the C-terminal C2-region, containing only random coil. The thermal unfolding of caldesmon takes place gradually without a steep transition and the unfolding is reversible upon cooling, consistent with the known "heat resistance" of caldesmon. This "continuum-of-states" unfolding contrasts with the sharp, cooperative, two-state unfolding characteristic of many proteins. The domains of caldesmon also unfold gradually with the degree of unfolding increasing in the order C-domain < intact molecule < central domain < N-domain, suggesting that the thermal stability decreases in this order. PMID- 8239660 TI - Antioxidant activity of all-trans-retinol in homogeneous solution and in phosphatidylcholine liposomes. AB - A kinetic quantification of the lipoperoxyl radical-scavenging activity of all trans-retinol has been carried out in homogeneous solution, when radicals were produced from the oxidation of methyl linoleate in methanol, initiated by the lipid-soluble 2,2'-azobis (2,4-dimethylvaleronitrile) (AMVN) as well as in a soybean phosphatidylcholine membrane model, in which peroxidation was induced either by AMVN or the hydrophylic 2,2'-azobis(2-amidinopropane)hydrochloride (AAPH). The physical microenvironment contributes to the determination of antioxidant efficiency of all-trans-retinol. In homogeneous solution the kinetic constant kinh is 3.5 x 10(5) M-1 s-1 and appears of the same order of magnitude as the inhibition constant measured for alpha-tocopherol under the same experimental conditions. Nevertheless, despite its very high chemical reactivity toward lipoperoxyl radicals, the overall antioxidant efficiency of all-trans retinol in this system appears quite limited, since the evaluated stoichiometric factor is 0.21. When the polyenoic chain of all-trans-retinol is incorporated into a phosphatidylcholine lipid bilayer, the antioxidant efficiency depends on the site of peroxyl-radical production. The highest lipoperoxyl radical scavenging activity is measured when radicals are generated by AHVN inside the bilayer multilamellar liposomes. Under these conditions, the relative antioxidant efficiency is similar to that of alpha-tocopherol, and the stoichiometric factor is 3.1. When radicals are generated by AAPH in the aqueous phase of an unilamellar liposomal system, the antioxidant effectiveness of all-trans-retinol appears reduced and lower than that measured with equivalent amounts of alpha tocopherol. Synergistic antioxidant effects between all-trans-retinol and alpha tocopherol are observed when both antioxidants are simultaneously incorporated into unilamellar liposomes in which peroxidation is induced by AAPH. This suggests that all-trans-retinol may interact with tocopheroxyl radicals, thereby regenerating alpha-tocopherol. This interaction, which may be related to molecular features and to the relative location of the antioxidants in the bilayer, could provide an effective antioxidant system that may be of great importance in vivo. PMID- 8239661 TI - Reactivity of plasma glutathione peroxidase with hydroperoxide substrates and glutathione. AB - We studied enzyme kinetics parameters of plasma glutathione peroxidase (GSHPx-P) and the major cellular enzyme, GSHPx-1, for the substrates, H2O2, linoleic acid hydroperoxide (LinOOH), and glutathione (GSH). The major objectives were to determine whether the relatively slow GSHPx-P enzyme had a lower reactivity with hydroperoxides or with GSH and to identify favored hydroperoxide substrates. The rate constants describing the reactivity of human GSHPx-P and human GSHPx-1 with LinOOH and H2O2 are in the same range; GSHPx-P is more reactive with LinOOH and GSHPx-1 is more reactive with H2O2. GSHPx-P also has a low level of reducing activity toward cholesterol 7 alpha-OOH and no detectable activity with the 5 alpha-OOH isomer in contrast to phospholipid hydroperoxide glutathione peroxidase (PHGPx) which readily reduced both isomers. GSHPx-P catalytic activity toward phospholipid hydroperoxides is demonstrable in the absence of detergents, enhanced at low concentrations by deoxycholate, and strongly inhibited by Triton X-100 and incorporation into liposomes. These properties are the opposite of PHGPx. These results suggest that GSHPx-P largely lacks the membrane interfacial properties of PHGPx. GSHPx-P exhibits a smaller GSH rate constant than GSHPx-1. This property partially explains the slower turnover of GSHPx-P with several hydroperoxide substrates; the low reactivity with GSH is not consistent with efficient GSHPx function in the bulk plasma volume. GSHPx-P kinetic properties suggest that it would function best as a free fatty acid hydroperoxidase in GSH rich microenvironments. Minimally, the secretion of reduced enzyme would permit it to scavenge free fatty acid hydroperoxides. PMID- 8239662 TI - Purification and characterization of amine oxidase from soybean seedlings. AB - A simple and rapid procedure for purification of soybean seedling amine oxidase is reported. The crude enzyme, obtained by ammonium sulfate fractionation was purified by ion-exchange chromatography on a cellulose phosphate column and batch affinity chromatography on 6-aminohexyl-Sepharose. Cyclohexylamine, a competitive inhibitor, was utilized to elute the enzyme. A homogeneous enzyme was obtained with a yield higher than 25%, the content of minor components being < or = 2%. The M(r) estimated by gel filtration is 113,000 and 77,000 by sodium lauryl sulfate-polyacrylamide gel electrophoresis. The enzyme is a dimer and contains two Cu2+ ion per molecule. Its EPR spectrum is typical of Cu2+ in a tetragonal symmetry. The enzyme oxidizes cadaverine at high rate, the specific activity being 4.3 mukat/mg. Molecular, spectroscopic, and kinetic properties of this enzyme are reported. PMID- 8239663 TI - Amino acid substitutions at position 73 in motif 2 of Escherichia coli alanyl tRNA synthetase. AB - Lysine73, located in the adenylate synthesis domain of Escherichia coli alanyl tRNA synthetase (AlaRS), was previously indicated to be an important residue for the interaction of this enzyme with the acceptor stem of its cognate tRNA (tRNA(Ala)). Replacement of this residue with glutamine produced a reduction in the catalytic efficiency of AlaRS in the aminoacylation assay, primarily through an increase in the apparent KM for tRNA(Ala) [Hill, K., and Schimmel, P. (1989) Biochemistry 28, 2577-2586]. Studies on the role of residue 73 in the interaction of AlaRS with its substrates have now been extended using the additional substitutions of asparagine, alanine, and glutamate. Analysis of each substituted enzyme in the ATP-PPi exchange and aminoacylation reactions reveals kinetic characteristics similar to those obtained with the glutamine substitution, except that the glutamate substitution causes a fivefold decrease in the affinity for alanine. These data verify that the positive charge on lysine 73, rather than its hydrophilic side chain, is of importance in the binding of the cognate tRNA, but do not support an ionic interaction of this residue with the RNA phosphate backbone. The collective data support the prediction that lysine73 is in motif 2 of AlaRS [Cusack, S., Hartlein, M., and Leberman, R. (1991) Nucleic Acids Res. 19, 3489-3498], but question the predicted alignment of this motif with other enzymes in its class. PMID- 8239664 TI - Expression of rabbit cytochromes P4504A which catalyze the omega-hydroxylation of arachidonic acid, fatty acids, and prostaglandins. AB - The omega-hydroxylation product of arachidonic acid is thought to be a potent vasoconstrictor or a precursor thereof in kidney. In this report, we have measured the capacity of four rabbit CYP4A enzymes, each expressed in COS-1 cells, to catalyze the omega-hydroxylation of arachidonic acid. These rates were compared to those obtained for other substrates such as lauric acid, palmitic acid, and prostaglandins PGE1 and PGA1. With the exception of P4504A5, all of the enzymes tested exhibited relatively high rates for the omega-hydroxylation of arachidonic acid. P4504A5 showed very little activity toward arachidonic or palmitic acids as compared to that toward lauric acid (< 10%). In contrast, P4504A6 and P4504A7 catalyzed the omega-hydroxylation of arachidonic acid at rates that were roughly 50% of that observed for lauric acid. P4504A4 was not active toward lauric acid, but it also catalyzed the omega-hydroxylation of arachidonic acid at a rate that was roughly 20% of that exhibited for PGE1. Thus, each enzyme exhibits a distinct substrate specificity profile across this panel of substrates. A sensitive RNase protection assay was used to provide a more quantitative estimate of the relative abundance of mRNAs encoding P4504A5, P4504A6, and P4504A7 in liver and kidney from control, pregnant, and clofibrate treated animals. CYP4A5 is the most abundant of the mRNAs, but it was not induced in kidney and only moderately (2-fold) in liver by clofibric acid. CYP4A7 exhibits a similar pattern of induction by clofibrate. In contrast, CYP4A6 is induced 12-fold in liver and 6-fold in kidney. The higher induction ratio largely reflects a lower basal level of expression for CYP4A6 than for CYP4A7 and CYP4A5. Following treatment with clofibrate, the amount of CYP4A6 mRNA is similar to those of CYP4A5 and CYP4A7. Pregnancy did not affect the expression of CYP4A5, CYP4A6, or CYP4A7, although it induced the expression of CYP4A4 to detectable levels in the liver and kidney, where it is not normally found in nonpregnant animals. Our results indicate that the enzyme whose mRNA is most highly induced by clofibric acid (P4504A6) and the enzyme selectively elevated during pregnancy (P4504A4) both exhibit relatively high rates for the omega-hydroxylation of arachidonic acid. PMID- 8239665 TI - Folate metabolism and deposition in eggs by laying hens. AB - White Leghorn hens were fed purified folate-deficient diets or commercial corn- and soybean meal-based diets supplemented with different amounts of folic acid. The folate contents of egg yolk and blood plasma from these hens were estimated with an isotope-dilution, radioligand-binding assay. Folates in egg yolk were concentrated approximately 43-fold relative to the blood plasma from which they were derived. Yolk and plasma folate concentrations became saturated with increasing dietary folate. Hens fed a commercial, folate-sufficient diet (0.72 mg folate/kg) produced eggs with slightly less than half of the maximal folate content. Based on tritium deposition in egg yolk and egg white, the biological half-life of [3H]folic acid injected intraperitoneally into two folate-sufficient hens was approximately 15 days, while it was > or = 40 days in two hens fed a purified folate-deficient diet (0.07 mg folate/kg) that also reduced egg production. Radioactivity in egg yolk was concentrated more than 100-fold relative to egg white in both cases. The [3H]folates remaining in the hens at the end of the experiment were substantially more concentrated in liver than in kidney, heart, or skeletal muscle. The specific radioactivity of folates in the liver of folate-deficient hens after 78 days was almost 10 times greater than in folate-sufficient hens after 39 days. Laying hens have highly efficient conservation and delivery systems for folates. PMID- 8239666 TI - Biosynthesis and degradation of meprins, kidney brush border proteinases. AB - Meprins, which are cell surface metalloendopeptidases, consist of two types of subunits, alpha and beta. Genetic factors determine which subunits and oligomeric forms of the enzyme exist in kidney in different strains of mice. In order to further explore factors that determine the concentration and activity of meprins, the rates of biosynthesis and degradation of the meprin subunits were determined in an organ culture system using ICR and C3H/He mouse kidneys. In biosynthesis experiments, the rate of incorporation of radiolabeled amino acids into immunoprecipitable forms of the alpha and beta subunits was determined. The rate of loss of radiolabel from the subunits was measured in pulse-chase experiments to determine the half-lives of the subunits. The rate of synthesis of the alpha subunit was twofold greater than that of the beta subunit: 861 +/- 32 vs 361 +/- 23 dpm/micrograms subunit protein/min for alpha vs beta. The rate of synthesis for total kidney protein in both strains was approximately 700 dpm/micrograms/min. The half-life for alpha was 8.9 +/- 0.24 h compared to 12.1 +/- 0.7 h for beta; the half-life for total protein in kidney was approximately 35 h. Thus, the half-lives of alpha and beta were similar and shorter than the half-lives of the average protein in kidney cells. The higher rate of synthesis of alpha is probably responsible for the greater abundance of this protein compared to beta in microvillus membranes. PMID- 8239667 TI - Evaluation of ligand exchange as a model of heterogeneity within the binuclear core of cytochrome c oxidase. AB - Dilution of detergent-solubilized cytochrome c oxidase has been found to increase the pH sensitivity of the a3-CuB site. Comparison of a large number of preparations, each purified at pH 8.8, showed either one or two exponential reactivity with cyanide, indicating that pH was not the only factor regulating kinetic heterogeneity within the binuclear core. Differences in heme a concentration during the soluble stages of purification were shown to have a substantial effect on the distribution of both optical and kinetic states within the isolated enzyme. Similar dilution effects were also inducible after purification. In particular, the apparent pKa of the Soret band maximum was shifted to higher pH following dilution of the enzyme. The resulting increase in the 414-nm conformer was also associated with enhanced conversion to the slow cyanide-binding form, suggesting that a single protonation event may give rise to both optical and kinetic changes. Two lines of evidence were contrary to this: First, by appropriate choice of experimental conditions it was possible to observe 30-50% of the cytochrome a3 centers in the 414-nm state with no associated evidence of the slow conformer. Second, the addition of formate at pH 8.8 induced conversion to both slow and 414-nm conformers, but the rates and extents of conversion were not correlated. Separate structural events are therefore proposed to modulate the optical and kinetic properties of the binuclear core. Since acidic pH has the well-established ability to induce both rapid-->slow and 430-->414-nm conversions, we suggest that the effects of dilution and formate are to mimic an acid jump by facilitating proton uptake into the binuclear core. PMID- 8239668 TI - Thermodynamics of reactions of nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate. AB - The thermodynamics of six reactions of nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate is discussed both from the viewpoint of the chemical equations and the biochemical equations for these reactions. Tables of the standard enthalpies of formation and standard Gibbs energies of formation of species are presented and are used to calculate standard enthalpies of reaction, standard Gibbs energies of reaction, and equilibrium constants K of chemical reactions at 25 degrees C, 1 bar, and ionic strengths of 0, 0.1, and 0.25 M. These tables are used to calculate standard transformed enthalpies of formation and standard transformed Gibbs energies of formation of reactants, standard transformed enthalpies of reaction, standard transformed Gibbs energies of reaction, and apparent equilibrium constants K' of biochemical reactions at 25 degrees C, 1 bar, pH 7, and ionic strengths of 0, 0.1, and 0.25 M. Since these reactions do not involve pK's of acid groups in the vicinity of pH 7, these reactions produce exactly 0, 1, or 2 mol of H+ per mole of reaction. The calculations are compared with experimental values. PMID- 8239669 TI - Physiological substrates for rat alcohol dehydrogenase classes: aldehydes of lipid peroxidation, omega-hydroxyfatty acids, and retinoids. AB - Alcohol dehydrogenase classes exhibit important differences in both substrate specificity and tissue distribution which suggest distinct physiological functions. We have studied the kinetic constants at pH 7.5 of the rat alcohol dehydrogenase classes, purified from liver (classes I and III) and from stomach (class IV), with three groups of relevant physiological compounds: cytotoxic aldehydes generated in lipid peroxidation, omega-hydroxyfatty acids, and retinoids. Classes I and IV actively reduce 4-hydroxynonenal, 2-hexenal, and hexanal, which are toxic compounds known to be produced in significant amounts during lipid peroxidation. Class III shows poor activity with these aldehydes. Class IV exhibits the best kcat/Km values (2150 mM-1 x min-1 for 4 hydroxynonenal), which suggest a role for this enzyme in the elimination of the cytotoxic aldehydes in tissues that are susceptible to lipid peroxidation, such as skin, cornea, and mucosa of the respiratory and digestive tracts, where class IV is localized. The three classes are very active with omega-hydroxyfatty acids, suggesting that all of them are involved in the physiological oxidation of these compounds in the rat tissues. The kinetic constants support that oxidation of omega-hydroxyfatty acids is a physiological function for class III, in addition to its role as formaldehyde dehydrogenase. Finally, classes I and IV are active in retinol oxidation and retinal reduction. Class IV may play a crucial role in the generation of retinoic acid in epithelia, where this compound is involved in development and cell differentiation. In conclusion, alcohol dehydrogenase is an enzyme with multiple metabolic roles, and the different substrate specificity and tissue localization for each class provide organs and tissues with distinct physiological functions. PMID- 8239670 TI - Decreased cytochrome oxidase activity and changes in phospholipids in heart mitochondria from hypothyroid rats. AB - The effect of hypothyroidism on kinetic characteristics of cytochrome oxidase in rat heart mitochondria was studied. Mitochondrial preparations from control and hypothyroid rats had equivalent Km values for cytochrome c, while the maximal activity of the oxidase was significantly decreased (more than 30%) in mitochondrial preparations from hypothyroid rats. This decrease is associated to a parallel decrease in state 3 respiration. The cytochrome aa3 content was slightly decreased (by around 15%) in mitochondria from hypothyroid rats. The Arrhenius plot characteristics differ for cytochrome oxidase activity in mitochondria from hypothyroid rats as compared with control rats in that the breakpoint of the biphasic plot is shifted to a higher temperature. Cardiolipin content was markedly decreased in the mitochondrial membrane from hypothyroid rats. No alterations were found in the pattern of cardiolipin fatty acid distribution of mitochondrial membrane from control and hypothyroid rats. The effects of the hypothyroid state on the activity of cytochrome oxidase, on cytochrome aa3 levels, and on cardiolipin contents were completely reversed by following the treatment of hypothyroid rats with thyroid hormone. The results support the conclusion that the depressed mitochondrial cytochrome oxidase activity in the hypothyroid state is due, at least in part, to a decrease in the cardiolipin content of the mitochondrial inner membrane. PMID- 8239671 TI - Hemin-induced lipid membrane disorder and increased permeability: a molecular model for the mechanism of cell lysis. AB - The interaction between hemin and lipid bilayers was examined from the point of view of: (a) changes in membrane permeability, (b) changes in lipid organization, (c) catalysis of lipid peroxidation. The leakage of solutes trapped in inner aqueous compartments of vesicles and liposomes increased in a concentration dependent manner, in the range of 100-1000 microM, saturating at ca. 400 microM hemin. Under saturating conditions, leakage was essentially complete after ca. 0.5 h. Antioxidants had practically no effect upon this behavior. Extensive leakage at 5 and 10 microM hemin was observed only in the absence of antioxidants and after ca. 10 h. Electron spin resonance spectra of a spin label incorporated in oriented lipid multibilayers indicated that hemin decreased the degree of order of the bilayer. The effect was instantaneous, time-independent, and unaffected by antioxidants, displaying a concentration dependence similar to that of the permeability studies. In contrast, hemin-catalyzed lipid peroxidation displayed a bell-shaped dependence on hemin concentration, the effect ceasing at concentrations lower than those required for instantaneous permeability and structural changes. The bell-shaped behavior was found both for the detection of thiobarbituric acid-reactive compounds (TBARS) and for oxygen consumption. The time scale of TBARS formation was much slower than that for permeability changes, significant amounts of products being detected only after leakage was essentially complete. Both permeability and lipid peroxidation occurred to a larger extent in cholesterol-containing membranes. Hemin is highly aggregated in aqueous phase. Upon binding to the membrane, the aggregates undergo dissociation in a hemin/lipid ratio-dependent manner. At low ratios, hemin is largely monomeric, while at high ratios, aggregates prevail. Monomers are more effective in promoting oxidation. Aggregates are responsible for the enhancement of permeability and membrane disorder. Oxidation of membrane components is often proposed to be involved in the mechanism of hemin-promoted cell lysis. Nevertheless, the role of oxidative processes is not clear in some hemoglobinopathies. The present results provide a molecular model for an alternative mechanism of lysis that would dominate in cases where the concentration is high enough for hemin to aggregate. PMID- 8239672 TI - [Autologous blood stem cell transplantation]. AB - Autologous blood stem cell transplantation (ABSCT) has been increasingly used, as the third generation of hemopoietic stem cell transplantation next to allogeneic and autologous bone marrow transplantation (allo-BMT and ABMT), in the treatment of malignant diseases. As to the principle of treatment, ABSCT is identical to ABSCT. However, ABSCT has some advantages over ABMT as follows: 1) a rapid engraftment, 2) no requirement of general anesthesia for harvesting hemopoietic stem cells, and 3) possibility of less contamination of tumor cells. Accordingly, ABSCT will take place of ABMT in the strategy of cancer treatment and improved results will be expected. PMID- 8239673 TI - [An outline of 5-HT3 receptor antagonists (1)--In pharmacological actions]. AB - This paper described an outline of pharmacological studies of 5-HT3 receptor antagonists, mainly on ondansetron, which control nausea and vomiting associated with cancer chemotherapy. Administration of cytotoxic drugs is known to increase serotonin (5-HT) concentrations, and when released, 5-HT provokes the emetic responses via two routes; 1) 5-HT acts as an intrinsic transmitter substance and stimulates the emetic response via 5-HT3 receptors on vagal afferent terminals; 2) 5-HT transmits impulses to the vomiting center via 5-HT3 receptors on the chemoreceptor trigger zone in the area postrema of the central nervous system. 5 HT3 receptor antagonists are thought to exert an antiemetic action by specifically and competitively blocking 5-HT3 receptors. PMID- 8239674 TI - [Current status and future prospects of the radiation oncology treatment planning system]. AB - Treatment planning is the most essential radiation oncology practice and has been developing owing to various technological innovations such as faster computing speed, advanced computer graphics and enormous data storage capacity. Treatment planning consists of CT data retrieval, target input, selection of treatment techniques and dose calculation, evaluation of dose distribution, and data transfer to the accelerator, each of which has been progressing quite rapidly. In particular, ongoing topics in treatment planning include multi-leaf conformation, non-coplanar technique, CT scanner dedicated to treatment planning and data transfer through LAN. Target delineation using PACS, and megavoltage verification imaging represented by portal imaging, are also useful. Higher local control rates as well as lower complication probabilities are expected with the sophisticated treatment planning system. PMID- 8239675 TI - [192Ir brachytherapy]. AB - Modern high technology has recently brought a precision treatment modality in the field of brachytherapy for cancer patients. Ir-192 manual afterloading (hair pin technique of Ir-192 wire) replaced the technique utilizing Ra-226 needles at our department in 1973. In May 1991, microSelectron-HDR (Ir-192 micro-source of 370 GBq) was installed in Osaka University Hospital. Preliminary analysis of phase I/II study resulted in no significant differences between the incidence of an acute mucosal reaction as well as early tumor response after high and low dose rate interstitial brachytherapy for oral cancer. Since April 1992, a phase III study has been under way to completely eliminate the problem of hospital personnel exposure to radiation in the field of brachytherapy. The introduction of remote after-loading of Ir-192 micro-source has resulted in improvements in elderly patient care during the interstitial brachytherapy for malignancies. The indications for HDR brachytherapy have been expanded, and new technology was developed to improve the local cure of the disease, such as linked double button technique for oral cancer, template interstitial brachytherapy for perineal cancer, and postoperative brachytherapy using intraoperative flexible catheter placement for locally unresectable disease or microscopically residual disease. Through these meticulous efforts, HDR interstitial brachytherapy will soon become a satisfactory substitute for traditional LDR interstitial brachytherapy. PMID- 8239676 TI - [Breast conservation treatment for breast cancer]. AB - Breast conserving treatment is defined as a combination of conservative surgery for resection of the primary tumor (lumpectomy, segmental mastectomy, or quadrantectomy), mostly with dissection of the axillary nodes, followed by radiation therapy for the eradication of residual microscopic disease in the breast and, in some cases, draining nodal tissues. The goal of this treatment is to provide highly satisfactory cosmetic results, without compromising local tumor control or survival compared with total mastectomy. This review article summarizes the results of prospective randomized trials comparing breast conserving treatment with mastectomy, the NIH consensus conference statement, patient selection, complications, cosmetic outcomes, the present situation of this treatment in Japan and our clinical results using CT simulator for radiotherapy treatment planning. PMID- 8239677 TI - [Gamma knife]. AB - The term "radiosurgery" was defined by Lars Leksell, a Swedish neurosurgeon, as the closed-skull destruction of a precisely defined intracranial target using high-dose radiation with stereotactic technique in a single session. For this purpose, the Gamma Knife was developed in 1968. It was equipped with multiple cobalt 60 sources and delivered collimated narrow radiation beams precisely concentrated to the focus. The dose gradient in the periphery of the treatment volume is extremely steep, so a high dose can be delivered to the small target volume sparing the surrounding normal tissue. Treatment planning was directed using a computer program developed for Gamma Knife and stereotactically obtained imaging database. The treatment procedure may be completed in only one day. The Gamma Knife makes it possible to treat deep-seated and surgically inaccessible lesions with low mortality and morbidity, and to control conventionally radioresistant lesions. Up to now, over 10,000 patients have undergone radiosurgery with a Gamma Knife around the world. The indications were primarily functional disorders, then expanded to include arterio-venous malformation (AVM), benign and a few malignant brain tumors. Excellent results were noted in the treatment of AVM. The two-year total obliteration rate of the nidus was 71-87%, and the adverse effect rate was 3-12%. The control rates of acoustic tumors was reportedly 85-89% with a lower incidence of facial nerve injury and a higher rate of hearing preservation. Gamma Knife radiosurgery has also been used to treat meningioma, pituitary adenoma and metastatic brain tumors. Its application for the treatment of malignant glial tumors or other tumors is developing. The capability of this technique is growing, and radiosurgery will be one of the important treatment modalities for selected neurosurgical or other pathological conditions. PMID- 8239678 TI - [Linac-knives]. AB - Stereotactic radiotherapy using linear accelerators (Linac-knives) is a promising approach for treatment of metastatic brain tumors, arterio-venous malformations, and benign brain tumors. The advantages of the linac-knife over the Gamma knife are its capacity for stereotactic fractionated radiotherapy and to extend the indication for head and neck tumors and cervical spinal tumors. PMID- 8239679 TI - [Proton radiotherapy]. AB - Protons have an ideal characteristic for radiotherapy in terms of dose distribution, and the clinical values of proton radiotherapy have been recognized over thirty years. The outstanding clinical results for the selected categories of patients such as uveal melanoma, sarcoma of the skull base and/or the cervical spine, and relatively slow-growing, intracranial lesions treated with proton therapy have already been well reported through many articles. In recent years, the technological development has produced proton accelerator that generates protons appropriate for the treatment of deep-seated tumors, and isocentric gantry system. It will be necessary however, to evaluate and establish the efficacy of proton radiotherapy for tumors that have not previously been studied during the next decade. The number of treatment facilities has been steadily increased over the world. Proton radiotherapy is expected to play an important role in cancer treatment in the near future. PMID- 8239680 TI - [Combination therapy with low dose adriamycin for advanced or recurrent breast cancer. Hokkaido Breast Cancer Treatment Study Group]. AB - Comparative clinical trials among 3 regimens for patients with advanced or recurrent breast cancer were performed as a multi-institutional joint study. Arm I of 3 regimens consisted of a 3-day consecutive administration of adriamycin (ADM) at 10 mg/body every 4 weeks, a daily oral administration of cyclophosphamide (CPA), 5-FU and tamoxifen (TAM) at 100 mg, 200 mg, and 20 mg, respectively, and a once-a-week intramuscular or subcutaneous injection of OK-432 (OK) gradually increased from the initial dose of 1 KE to the maintenance dose of 5 KE. Arm-II contained methotrexate (MTX) at 10 mg/body for 3 consecutive days every 4 weeks in place of ADM in Arm-I. Arm-III contained neither ADM nor MTX. Of 69 cases registered, 52 were eligible, leaving 6 non-eligible and 11 incomplete cases. The results of the overall evaluation of 48 complete cases other than 4, in which the tumor was not exactly measured, were as follows. Arm-I resulted in 2 CR and 3 PR out of 19 cases, the response rate being 23.6% (5/19). Arm-II resulted in 1 CR out of 15, the response rate being 6.7% (1/15). In Arm-III, no response cases were found. No significant difference was observed among three treatment groups (p = 0.055), but in Arm-I, the response rate was higher than in the other treatment groups, suggesting that there is a probability of useful combined use of ADM. PMID- 8239681 TI - [Chemo-endocrine therapy with high dose medroxyprogesterone acetate for recurrent or advanced breast cancer]. AB - The endocrine therapy with high-dose medroxyprogesterone acetate (MPA) was applied for 19 patients with recurrent breast cancer and 3 with advanced breast cancer. According to the dominant lesion of the disease, the patients were divided into three groups; 13 with soft tissue lesion, 5 with visceral lesion, and 4 with osseous lesion. The serum MPA level was higher than 25 ng/ml in 14 cases out of 19 examined. The results of objective responses in second-line therapy with MPA were CR in two cases, and NC or PD in 20 cases. In 19 out of 22 patients with MPA therapy, the periods of survival were longer than the period of 50% survival of the patients without MPA therapy. PMID- 8239682 TI - [Correlations between estrogen receptor and prognostic factors of patients with breast cancer. The Kanto Cooperative Study Group of Adjuvant Chemoendocrine Therapy for Breast Cancer, Japan]. AB - We investigated possible correlations between estrogen receptor (ER) and prognostic factors in 3,789 patients with stage II or stage IIIa breast cancer in Japan. The patients with ER-positive tumors showed a significantly better prognosis in the 5-year survival rate and the 5-year disease-free survival rate compared with the patients with ER-negative-tumor. Positivities increased with increasing patient age, with increasing tumor size, and lowering degree of cell differentiation, and no relation was observed between the positivities and tumor sites, obesity index, and lymph-node metastasis. PMID- 8239683 TI - [Concomitant arterial infusion chemotherapy with tamoxifen therapy for hepatic metastases from pancreatic adenocarcinoma--a preliminary report]. AB - The prognosis for hepatic metastasis from pancreatic adenocarcinoma is generally poor. Estrogen receptors have recently been demonstrated in pancreatic adenocarcinoma, and it has been suggested that tamoxifen increases patient survival. We have tried tamoxifen therapy combined with intermittent intraarterial infusion chemotherapy using mainly 5-FU for 4 patients with hepatic metastasis from pancreatic adenocarcinoma. Clinical evaluation of initial response was available for these 4 patients. Although the number of the patients in this study was small, we obtained a response rate of 50%; CR in one case, PR in one case. No patients showed progressive disease (PD). Median survival period was 4.6 months; 3 of 4 patients died with carcinomatous peritonitis at 2.5 to 6 months from the initiation of the therapy. No severe complication was observed. It is believed that this new concomitant therapy is effective in controlling hepatic metastasis from pancreatic adenocarcinoma. But additional therapy for carcinomatous peritonitis, which often develops after achieving hepatic control, is necessary to achieve long-term survival benefit. PMID- 8239684 TI - [Alternating CHOP-MEVP chemotherapy for advanced-stage, intermediate- and high grade non-Hodgkin's lymphomas]. AB - Twenty-one patients with advanced-stage, intermediate- and high-grade non Hodgkin's lymphomas were treated with alternating CHOP-MEVP chemotherapy. CHOP therapy consisted of CPA 650 mg/m2, ADM 45 mg/m2, VCR 1.4 mg/m2 and Pred 40 mg/m2 (po). MEVP therapy consisted of MIT 10 mg/m2 (iv) VDS 2 mg/m2 (iv) on day 1, etoposide 200 mg/m2 (po) on days 1-3, and Pred 40 mg/m2 (po) on days 1-5. Three courses of CHOP therapy and MEVP therapy were alternatively administered every three weeks. CR was achieved in 15 (71.4%) of 21 patients. Survival rate and relapse-free rate at 2 years for all 21 patients were 61.9% and 30.9%, respectively. Toxicity was generally tolerable except for CMV interstitial pneumonitis in a patient with IBL-like T-cell lymphoma and secondary leukemia in a patient with T-cell lymphoma. Chemotherapy of higher dose intensity is required to improve the relapse-free survival rate in these subsets of lymphoma. PMID- 8239685 TI - [Some problems of TS measurement after administration of fluoropyrimidines in colorectal cancer. Kinki Cooperative Study Group of Chemotherapy for Colorectal Carcinoma]. AB - A joint multicenter comparative study of carmofur (HCFU) alone or HCFU+dipyridamole (DP) concurrent administration was conducted by the Kinki Colorectal Cancer Chemotherapy Society. In this study, the drugs were administered five days prior to surgery, and thymidylate synthetase (TS) inhibition rate at the primary lesion of colorectal cancer was determined. In addition, the effect of time after resection of the primary lesion before specimen cooling on the TS inhibition rate was investigated. The subjects were 87 registered patients who received colorectal cancer surgery during 4 months from October 1991 to January 1992. The time before postsurgical cooling was divided into four group; within 10 min (n = 67), 11-20 min (n = 8), 21-30 min (n = 7), and 31 min or longer (n = 5). The TS inhibition rates (mean +/- SD) were 0.45 +/- 0.22, 0.38 +/- 0.19, 0.30 +/- 0.21, and 0.04 +/- 0.05 respectively. The longer the time after resection of the primary lesion before specimen cooling, the more TS inhibition rate decreased. When the time before specimen cooling exceeds 30 min, the major part of TS combined with FdUMP exists in a free state. Therefore, it was considered not suitable as the material for the measurement. PMID- 8239686 TI - [A case of locally advanced breast cancer successfully treated with intra arterial infusion of high-dose epirubicin]. AB - A 43-year-old female with locally advanced breast cancer was treated with preoperative intra-arterial infusion chemotherapy using an implantable port catheter. The therapeutic regimen was comprised of two cycles at 3-week intervals. One cycle consisted of 50 mg of epirubicin which was administered on day 1, 4 and 7. A remarkable loco-regional response was confirmed only after two repeated cycles of the regimen. The side effects such as hair loss, general fatigue and leukopenia (nadir 1,700) were encountered, but these were moderate and had no influence on the patient's quality of life. These findings suggested that intra-arterial infusion of high-dose epirubicin via an implantable system was an efficient modality for the treatment of breast cancer. PMID- 8239687 TI - [A case of advanced hepatocellular carcinoma, in which the tumor almost disappeared by orally administered UFT]. AB - A 59-year-old male with liver cirrhosis was admitted to our hospital for further examination of general fatigue. A huge tumor was found in the right lobe of his liver with abdominal ultrasonography and computed tomography (CT). The titers of serum AFP (14,055 ng/ml) and PIVKA-II (more than 50.0 AU/ml) were extremely high. A tumor embolus was observed in the portal vein with abdominal angiography. According to these findings, he was diagnosed as having advanced hepatocellular carcinoma (HCC). He was treated by oral administration of UFT (600 mg/day) as an unresectable case of HCC. Three months later, the clinical symptoms were greatly improved and HCC was almost diminished with abdominal ultrasonography and CT, although a small tumor embolus was found by angiography. The titers of AFP and PIVKA-II were reduced to the normal range. This case suggests the clinical effectiveness of UFT for the treatment of HCC. PMID- 8239688 TI - [Vanishing tumor thrombus in IVC after oral administration of UFT and TAE in a patient with HCC; a case report]. AB - A 74-year-old man was diagnosed as hepatocellular carcinoma (HCC) in the area of S6-7 with tumor thrombus continuously developed from hepatic vein to inferior vena cava (IVC). AFP value remained high despite two transcatheter arterial embolizations with adriamycin, mitomycin and Lipiodol (Lp-TAE). The AFP value gradually decreased two weeks after oral administration of UFT. Nine months after, the AFP level was 26.4 ng/ml and the tumor thrombus in the IVC disappeared. The size of the main tumor did not change. This case suggests Lp-TAE and UFT are useful for suppressing the growth of the main tumor and to extinguish the tumor thrombus. PMID- 8239689 TI - [A case of icteric type hepatoma responding to MMC infusion therapy via cannulation into the hepatic artery]. AB - Hepatoma rarely presents with obstructive jaundice. We describe a case of hepatoma, a 66-year-old woman, presenting with obstructive jaundice, whom we treated with intra-arterial infusion of the anticancer drug, MMC. The hepatic main tumor was situated in the border between the medial and lateral lobe, which caused an obstruction of the common hepatic duct, accompanied with multiple intrahepatic liver metastases. MMC was injected weekly into the reservoir of the Infuse-A-Port, which was located subcutaneously. Intra-arterial infusion therapy reduced the tumor size and tumor markers. PMID- 8239690 TI - [A case of complete remission of metastatic skin carcinoma (erythema type) from advanced gastric cancer by CDDP administration]. AB - A 75-year-old female with skin metastasis from advanced gastric cancer (portion A, tub2, ss beta, n2, P0H0, Borrmann 3-type) was successfully treated with CDDP at a dosage of 25 mg/body every week or two. She had undergone R2 curative subtotal gastrectomy in October 1986, and had orally taken tegafur 300 mg/day as postoperative adjuvant chemotherapy. Three years after the operation, she had skin erythema at chest and neck. A pathological examination revealed that the skin erythema was a metastasis of gastric cancer. After administration of 5-FU and MMC, the skin lesion diminished in size, but in November of 1990, it expanded again. Administration of 5-FU, MMC and epirubicin was not effective. The skin lesion expanded extensively and changed to nodular type. CDDP was intermittently administered from August 1991 to December 1991, 25 mg/body div. every week or two; the total volume of CDDP reached 225 mg. The metastatic skin carcinoma disappeared and a complete response was obtained. CEA, which had been high (190 ng/ml), regained the normal level. Thrombocytopenia and liver dysfunction developed as side effects, but reduced after cessation of CDDP administration. The patient has been well for 3 years and 6 months since the onset of the skin metastasis. PMID- 8239691 TI - [A case of gastric malignant lymphoma markedly responding to chronic daily and oral administration of low-dose etoposide]. AB - A 71-year-old woman was diagnosed as malignant lymphoma (non-Hodgkin's lymphoma, diffused mixed cell type) which involved gastric lesions. Combined chemotherapies were undertaken using VCR, CPA, 4-epi-ADM and PRED and then etoposide, 4-epi-ADM and PRED. After about 5 months, recurrent lesions were revealed in the right supraclavicular lymph node, so we conducted additional chemotherapy. However, the patient rejected intravenous therapy and required out-patient therapy, so daily oral administration of etoposide (50 mg/day) was carried out. Because of the toxicities of alopecia, the dose was reduced to 50 mg every other day, then to 25 mg every other day. No recurrent signs have been seen for the past 3 years. Bone marrow suppression was little. Chronic oral administration of low-dose etoposide is the potential salvage therapy of gastric malignant lymphoma. PMID- 8239692 TI - [Neoadjuvant chemotherapy effective for small cell lung cancer in a nephrectomized patient]. AB - A 67-year-old man, who was nephrectomized due to renal cell cancer 4 years ago, was admitted to examine a mass shadow in the right middle lung field. He was diagnosed as small cell lung cancer with TBLB. Because of impaired renal function, he was treated with CBDCA (300 mg/m2, day 1), THP (30 mg/m2, day 1) and oral etoposide (25 mg/body, for 21 days) without any renal complications. After 3 courses of chemotherapy, the lung CT showed scar lesion despite the disappearance on the chest X-ray, and a right lower lobectomy was performed. Malignant cells remained in the scar lesion, but not in the lymph nodes. These findings suggested the effectiveness of neoadjuvant chemotherapy. This newly-designed chemotherapy procedure is necessary for patients with renal complications. PMID- 8239693 TI - [A case of pulmonary metastasis from colon cancer successfully treated by 5-FU combined with leucovorin and interferon alpha-2a]. AB - A 58-year-old woman with colon cancer, who had received oral 5-FU over 17 months after right hemicolectomy, was diagnosed as having a recurrence of the disease with multiple pulmonary metastasis. She was treated for 5 days with a combination of continuous infusion of 5-FU 600 mg/m2/day, bolus injection of leucovorin (LV) 20 mg/m2/day, and intramuscular injection of interferon (IFN)-alpha-2a (6.0 x 10(6) U/day, repeated every 3 weeks. The chest X-ray after three cycles showed a decrease in size of metastatic lesions by 51%, indicating a partial response. Correspondingly, the serum levels of CEA and CA 19-9 significantly decreased. There were modest but tolerable side effects such as fever, nausea, diarrhea, stomatitis, and alopecia. The patient has been given oral UFT and LV after discharge, and is still alive with continued improvement of pulmonary lesions even 9 months after initial chemotherapy. Although the detailed synergistic mechanism of 5-FU and IFN has yet to be determined, the addition of IFN, as a biochemical modulator distinct from LV, to the combination of 5-FU and LV, appears to further potentiate the therapeutic efficacy and may be useful for advanced colorectal cancer. PMID- 8239694 TI - [Recombinant human erythropoietin in the treatment of the anemia of gynecological tumor patients]. PMID- 8239695 TI - [Antiproliferating effect of HMG-CoA reductase inhibitor on human pancreatic cancer cell]. PMID- 8239696 TI - [In vitro effect of 1-hexylcarbamoyl-5-fluorouracil on human cancer cell lines of gastrointestinal tract]. PMID- 8239697 TI - [Usefulness of BFP for screening of early urinary bladder cancer]. PMID- 8239698 TI - Mendelism in early 20th-century American dermatology. AB - Now, almost a century ago since Mendel's work was rediscovered, is a propitious time to review the appearance of mendelian concepts in American dermatology. In our world of instantaneous and sometimes overwhelming dissemination of new information and the apparent rapid incorporation of new knowledge into our specialty, it is useful to examine how heredity was incorporated in the dermatologic literature in the first 25 years of the 20th century. The dermatologic literature serves as a surrogate for the thought of the era. PMID- 8239699 TI - Principles of genetics in the molecular era. A primer for dermatologists. AB - Advances in technology have resulted in an explosion of information about the molecular bases of human inheritance and human disease. Much of the new terminology is daunting, and it is becoming increasingly difficult to remain current. This article briefly reviews some of the changes in our understanding of the rules of inheritance and recurrence risks for genetic disorders. Further readings are suggested, and a glossary of technical terms is provided. PMID- 8239700 TI - The morbid cutaneous anatomy of the human genome. AB - BACKGROUND: The identification of genes whose mutations underlie hereditary diseases has long been a holy grail and one whose satisfaction very recently has become a reality for a surprisingly large number of dermatologic diseases. OBSERVATIONS: This review summarizes in brief the chromosomal localization and, where known, the gene identification for diseases expressed primarily in the skin. These vary from skin cancer genes whose products control cell growth and carry out DNA repair to skin fragility genes whose products impart mechanical stability to the skin. CONCLUSIONS: The explosion of new data in the past several years summarized herein is nearly unprecedented and bodes well for the development of new therapeutic approaches to common as well as rare skin disorders. PMID- 8239701 TI - From the clinic to the research laboratory. The role of the clinician in molecular genetic studies. AB - BACKGROUND: The first physician to examine a patient with a genetic disorder or birth defect is usually a specialist in a field other than genetics. The presentation of certain categories of patients of particular interest to molecular genetics research may be distinct. The recognition of these patients by clinicians is fundamental to the study of genetic disorders at the DNA level. OBSERVATIONS: Neurofibromatosis type 1 is a paradigm for how the study of a single genetic disease and its multiple molecular features has been facilitated by the use of various categories of patients. Other examples of interest to dermatologists, surgeons, and other specialists are discussed to demonstrate how the identification of key patients was instrumental in studies of gene localization and subsequent cloning, gene clusters or contiguous gene deletion syndromes, or mutation phenomena such as imprinting, uniparental disomy, and gonadal mosaicism. The molecular researcher has limited access to surgical specimens, and the donation of skin, tumor, and other tissues may lead to increased knowledge of new mutations in somatic mosaicism, or loss of heterozygosity of tumor suppression genes in cancer. CONCLUSIONS: Guidelines are suggested to alert the physician to each of these categories of individuals with unusual presentation, as well as to recognize that the study of families with rare disorders may enable scientists to locate the responsible genes. The teamwork of clinician and molecular researcher is essential for the advancement of our understanding of DNA mechanisms in genetic disease. The ethics involved in referral of patients to molecular genetic research studies are discussed. PMID- 8239702 TI - Prenatal diagnosis of genetic skin disease using fetal skin biopsy samples. AB - BACKGROUND: Understanding normal skin development and identifying markers of genetic skin disease expressed in postnatal skin have permitted the prenatal diagnosis of many severe genodermatoses: bullous diseases, keratinization diseases, pigment cell disorders, and disorders of the epidermal appendages (ectodermal dysplasias). Samples of 16 to 22 weeks' gestation fetal skin obtained by ultrasound-guided biopsy are evaluated using morphologic, immunohistochemical, and biochemical methods. OBSERVATIONS: The 12-year experience in evaluating samples from fetuses at risk of these disorders has allowed us to establish conditions that must be met before the samples are taken and the criteria for recognizing the disorder, to recommend the site(s) for sampling, and to be mindful of pitfalls that may be encountered in interpreting the tissue structure. CONCLUSIONS: Fetal skin biopsy is an important diagnostic tool that has permitted families in which members carry the abnormal gene for one of these severe skin diseases to undertake a pregnancy knowing that the condition of the fetus can be determined. Nonetheless, the ultimate goal is phase out this procedure when linkage of more of these disorders to specific genes is understood, specific mutations are characterized, and probes are available for molecular diagnoses using tissue obtained at earlier fetal ages. Until this is possible, fetal skin biopsy remains an important tool that can be used with reasonably high levels of safety and confidence. PMID- 8239703 TI - Mosaicism in human skin. Understanding the patterns and mechanisms. AB - BACKGROUND: The skin is especially suitable for the study of mosaicism. In this review, the various genetic mechanisms leading to mosaicism and the resulting cutaneous patterns are considered. OBSERVATIONS: Mosaicism may produce different cutaneous patterns such as the lines of Blaschko, the checkerboard pattern, the phylloid pattern, and a patchy pattern without midline separation. A unique lateralization pattern is observed in the CHILD syndrome. Two major genetic categories are functional mosaics resulting from X inactivation and genomic mosaics caused by autosomal mutations. Functional mosaicism may be caused by either male-lethal or nonlethal X-linked mutations. Similarly, autosomal mutations resulting in genomic mosaicism may be either lethal or nonlethal. Many mosaics are caused by loss of heterozygosity, and uncommonly this mechanism may give rise to twin spots such as vascular twin nevi. Some cutaneous mosaic phenotypes virtually always occur sporadically, but exceptionally may show a familial aggregation. This paradox may be explained by paradominant inheritance. Heterozygous individuals are, as a rule, unaffected, but they express the birthmark when allelic loss occurs during embryogenesis. CONCLUSIONS: The concept of cutaneous mosaicism is important for gene mapping because here we have the opportunity to study two populations of cells differing only with regard to the mutation causing mosaicism. Future research will probably show that a specific genetic anomaly, when present as a mosaic, always produces the same type of cutaneous pattern. PMID- 8239704 TI - Keratinocyte gene therapy. AB - BACKGROUND: Gene therapy is currently being used in clinical trials to treat a variety of diseases. In keratinocyte gene therapy, the gene that will correct the disease by expressing the normal protein or enzyme is inserted and expressed in keratinocytes. Keratinocytes have significant potential, as a target cell of gene therapy, in the treatment of both systemic diseases as well as skin diseases caused by a genetic defect in keratinocytes. OBSERVATIONS: Although keratinocyte gene therapy is not yet being tested in clinical trials, animal models do exist where keratinocytes are being used to secrete factors such as human growth hormone and factor IX (for hemophilia) into the systemic circulation. Genetic diseases of the skin such as recessive epidermolysis bullosa dystrophica or xeroderma pigmentosum have not yet been treated with keratinocyte gene therapy in animal models. CONCLUSIONS: Keratinocytes have many advantages as a target cell in gene therapy, and progress has been made using animal models. However, the sustained and efficient delivery of factors to the bloodstream by keratinocytes expressing a transgene has not yet been accomplished. Future goals are to obtain adequate levels of the desired factors, hormones, or enzymes for sustained periods of time, either in keratinocytes or in the vascular system. PMID- 8239705 TI - Gene rearrangements and T-cell lymphomas. AB - BACKGROUND: Cutaneous T-cell lymphomas comprise a broad spectrum of neoplasia ranging from indolent to highly aggressive types. To determine subset lineage and malignant vs benign nature, morphologic analysis, immunophenotyping, and flow cytometry have been used. However, given the shortcomings of these methods, molecular genetic techniques, which take particular advantage of the clonal nature of malignancy, are now being applied to better characterize and diagnose these lymphomas. RESULTS: Each antigen-specific T cell and its clonal progeny has a unique rearrangement of its T-cell receptor gene such that it can recognize very specific antigenic epitopes. By visualizing these particular T-cell receptor gene rearrangements, Southern hybridization techniques and polymerase chain reaction amplification can detect clonal populations of T cells in the skin, blood, and lymph nodes of patients with T-cell leukemias and lymphomas. Clonal T cell populations have also been found in cases of benign disorders such as lymphomatoid papulosis and pityriasis lichenoides et varioliformis acuta. Although these disorders usually have a benign outcome, they may represent dysplastic clonal lymphoid expansions with a high incidence of spontaneous regression. CONCLUSIONS: Molecular genetic techniques have added to our ability to diagnose, characterize, and monitor the course of T-cell lymphomas and leukemias. In addition, they may provide insight into the pathogenesis of certain benign disorders. PMID- 8239706 TI - Molecular diagnosis of lymphocytic infiltrates of the skin. AB - BACKGROUND: Advances in our understanding of the molecular genetics of lymphocyte antigen receptors (B-cell immunoglobulin and T-cell antigen receptor), have led to the application of molecular biologic techniques to molecularly characterize lymphocytic infiltrates of the skin. Molecular diagnosis refers to the application of these techniques as a diagnostic aid in the clinicopathologic evaluation of cutaneous lymphocytic infiltrates. OBSERVATION: Molecular studies have clinical application in the determination of lineage and detection of retroviruses in cutaneous lymphoid neoplasms, distinguishing between lymphoproliferative and reactive infiltrates, and staging and monitoring response to therapy in cutaneous T-cell lymphoma. Southern blot analysis of immunoglobulin and T-cell antigen receptor gene rearrangements may fail to aid the clinician in establishing a diagnosis of a cutaneous malignancy due to the limits of detection sensitivity in minimally infiltrated lesions (eg, parapsoriasis and patch-stage mycosis fungoides) or the still uncertain prognostic significance of clonality in benign cutaneous diseases (eg, follicular mucinosis, pityriasis lichenoides et varioliformis acuta, lymphomatoid papulosis, and cutaneous lymphoid hyperplasia). CONCLUSIONS: Molecular studies have enormous research value, providing new means to explore the pathogenesis and clonal evolution of lymphoproliferative skin diseases. Presently, however, they have limited applications as an independent diagnostic tool. As our understanding of the clinical and biologic significance of the molecular detection of clonal lymphocyte populations in the skin expands and as the application of polymerase chain reaction amplification provides us with greater detection sensitivity and specificity, the clinical utility of molecular diagnosis of lymphocytic infiltrates of the skin will be enhanced. PMID- 8239707 TI - Tender bluish papules on the trunk and extremities. Blue rubber-bleb nevus syndrome. PMID- 8239708 TI - A family with lesions on the face, hands, and buccal mucosa. Cowden's disease. PMID- 8239709 TI - Asymptomatic slowly growing tumor on the forearm. Neurothekeoma. PMID- 8239710 TI - Asymptomatic papule on the lower leg. Clear-cell acanthoma. PMID- 8239711 TI - Gut contents: a significant contaminant of Mytilus edulis whole body metal concentrations. AB - Ingested matter can have a significant effect on whole body metal concentration measurements in Mytilus edulis. Depuration of mussels in clean seawater for 36 h prior to dissection eliminates most of these contaminating gut contents. Depuration followed by metal analyses is the most direct method of determining mussel tissue metal bioburdens. After being transplanted into a plume of primary treated sewage effluent in Salem Harbor, Massachusetts for 32 days, Al, Cr, and Fe concentrations in depurated mussels were significantly lower than those determined for either non-depurated mussels or for depurated mussels to which fecal concentrations of Al, Cr, and Fe were added back in. Although mathematical methods developed by both Ouellette (1978) and Boehm et al. (1988) could be applied to non-depurated mussels in order to correct for errors associated with gut metal contamination, these indirect methods were not as reliable as depuration prior to analysis. PMID- 8239712 TI - Mercury and major essential elements in seals, penguins, and other representative fauna of the Antarctic. AB - Concentrations of total Hg and major essential elements, Ca, Mg, Na, and K, were measured in the muscle, liver, and kidney of three species of seals, crabeater seal (Lobodon carcinophagus), leopard seal (Hydrurga leptonyx), and Weddell seal (Leptonychotes weddelli) collected in the Antarctic. The muscle and liver of three species of penguins, i.e., gentoo penguin (Pygoscelis papua), Adelie penguin (P. adeliae), chinstrap penguin (P. antarctica), and other representative fauna were also analyzed for the elements. Distinct inter-tissue differences in the metal concentration were observed; liver had the greatest concentrations of Hg, kidney showed maximum concentration of Ca and Na, while muscle was characterized by the greatest content of Mg and K. Inter-specimen differentiation of concentrations among the same species was distinctly visualized for Hg but not for the major essential elements. The Hg concentration in the seals analyzed are in keeping with those reported previously by other authors. Such comparison for Ca, Mg, Na, and K was impossible because of the lack of available literature data for their concentrations in marine seals. Numerous significant correlations were observed between concentrations of the several metals analyzed. There was no correlation between Hg, which is a non-essential element and both Zn and Cd which are essential and non-essential elements, respectively in the liver or kidney of seals. However, there were significant correlations between concentration of Zn and the sum of molar concentrations of Hg+Cd in kidney (r = 0.82) and liver (r = 0.76). The results suggest that several control mechanisms operate to maintain physiologically required levels that decreases any effect of heavy metal toxicants such as Hg and Cd. PMID- 8239713 TI - Muskrat populations in Virginia's Elizabeth River: physiological condition and accumulation of environmental contaminants. AB - This study evaluated the physiological condition and environmental contaminant concentrations in muskrats inhabiting the contaminated lower region of the Elizabeth River, Virginia, (USA). Muskrats trapped in the lower region of the Elizabeth River weighed less, had lower mean fat indexes, lower relative spleen weights, greater relative adrenal weights, and an increased incidence of disease and parasitism compared to muskrats trapped in the less contaminated upper region of the Elizabeth River or Nansemond River. Concentrations of the metals aluminum, cadmium, copper, nickel, and zinc and the number of polyaromatic hydrocarbon compounds were greater in lower Elizabeth River muskrats compared to upper region Elizabeth River or Nansemond River muskrats. These physiological data indicated that muskrats in the lower region of the Elizabeth River were in poor health relative to muskrats from the other regions studied. However, no significant difference in fecundity or density was observed among regions studied, suggesting that the level of environmental contaminant exposure was not sufficient to elicit a population level response. Greater population declines following severe natural environmental stresses (large storms, extreme cold, etc.) would be anticipated in lower region Elizabeth River muskrats; however, the muskrat population in this region does not now appear to be at risk of extirpation. PMID- 8239714 TI - Sublethal effects of three pesticides on Japanese medaka. AB - One- to 2-day-old medaka (Oryzias latipes) larvae were exposed for 4 days to the rice field pesticides methyl parathion, molinate, carbofuran and a mixture of all three. Pesticide concentrations were one-half the 96 h LC50 ("high concentration") and levels approximating those measured in receiving waters from rice field runoff ("low concentration"). Maximum swimming speed, spontaneous muscular activity, acetylcholinesterase activity, dry weight, RNA:DNA ratio, and five morphometric variables were determined at the end of the exposures. Larvae were retained for an additional 10 days in non-contaminated water, and the same measurements taken to investigate residual effects. Results are compared to a parallel study on striped bass larvae to evaluate the suitability of this species as a surrogate for the bass in toxicological studies involving sublethal exposures. There was no relationship between mortality rate and pesticide exposure either during the exposures or during the ten day subsequent period. Only the high concentration of carbofuran caused an impairment of swimming performance. Spontaneous activity was stimulated in the high concentration of molinate and the combined pesticides groups. Acetylcholinesterase was severely inhibited in parathion and molinate, and this persisted in some cases after 10 days in non-contaminated water. The pesticides had little effect on growth rate except for molinate which acted as a stimulant. Combining the three pesticides caused a less than additive effect. Except for decreases in acetylcholinesterase, the sublethal effects of the pesticides tested at the very low concentrations used were subtle. Apparently, larvae of this species are less sensitive to these pesticides than are striped bass larvae. PMID- 8239715 TI - In vitro immunomodulation by pentachlorophenol in phagocytes from an estuarine teleost, Fundulus heteroclitus, as measured by chemiluminescence activity. AB - Chemiluminescence (CL) was used to assay effects of pentachlorophenol (PCP) on the production of reactive oxygen intermediates (ROIs) generated by leukocytes from an estuarine teleost, Fundulus heteroclitus. Two parameters were measured during the phagocytically-induced respiratory burst: peak CL (maximal value) and total CL (temporal summation of induced values). Phagocytes obtained from the pronephros were incubated in the presence of sublethal doses of analytical (A-) or technical (T-) grades of PCP. Technical PCP has been reported to be more highly immunosuppressive than A-PCP in mammals; this effect was attributed to dioxins and other contaminants produced during manufacture. In this fish, both grades of the compound produced significant dose-dependent inhibition of peak and total CL. However, there were few significant differences between the effects of the two PCP grades on the magnitude of the CL responses at identical, sublethal concentrations, although T-PCP did produce a more marked reduction in the relative level of leukocytic CL as a function of dose than did A-PCP. Because of the probable roles of ROIs in antimicrobial blood cell-mediated mechanisms, reduced CL activity was interpreted as an indication of xenobiotic-induced immunosuppression. PMID- 8239716 TI - Organochlorines including polychlorinated biphenyls in muscle, liver, and ovaries of cod, Gadus morhua. AB - Twenty-three specific organochlorine contaminants and polychlorinated biphenyls (PCBs), measured as three Aroclor standards were analyzed in muscle, liver, and ovaries of cod, Gadus morhua, collected in the Northwest Atlantic. In general, contaminants were undetectable in muscle tissue, while concentrations were 10 times lower in ovaries than liver (wet weight). Comparison of results to other locations indicated a similarity between the ratio of the concentrations of p,p' DDE, p,p'-DDD and p,p'-DDT, in liver of cod from the northern North Sea and from the Northwest Atlantic, although with lower levels in the present study. The ratio of alpha-HCH and gamma-HCH was between that of the central and northern North Sea. Similar ratios tend to indicate similar residence times in the atmosphere, from source to sampling area. Comparison of sigma PCB and sigma DDT in the liver of cod from various geographical locations showed the following general trend in concentrations: Arctic, Northwest Atlantic, West Atlantic, Norway < North Baltic, Nova Scotia, North Sea < South Baltic. It was observed that if the liver concentration of one compound was low (high), there was a tendency for all compounds to be low (high). Cluster analysis of organochlorines in liver pointed to the presence of four basic clusters, which could reflect similar physical chemical properties within a group. Concentrations of organochlorines in ovaries were below levels expected to affect egg and larval viability. PMID- 8239717 TI - Effects of arsenic on DNA synthesis in human lymphocytes. AB - Effects of arsenic on DNA synthesis in human lymphocytes were biphasic: Either trivalent (arsenic trioxide and sodium arsenite) or pentavalent (sodium arsenate) arsenic compounds at very low concentrations enhanced DNA synthesis in human lymphocytes stimulated by phytohemagglutinin (PHA), whereas higher concentrations inhibited DNA synthesis. There were differences among individual susceptibilities to arsenic-induced DNA synthesis. Either stimulating or inhibiting effects of trivalent arsenic on DNA synthesis in PHA-stimulated lymphocytes were always stronger than those of pentavalent arsenic. Both trivalent and pentavalent arsenic could be rapidly taken up into the human lymphocytes and immediately stimulate or inhibit DNA synthesis. A possible dual effect of arsenic at very low concentrations as both comutagen and inhibitor of mutagenesis is discussed. PMID- 8239718 TI - Protection afforded greenhouse pesticide applicators by coveralls: a field test. AB - Applicators of chlorpyrifos, fluvalinate, and ethazol to ornamentals in a Florida greenhouse were monitored for exposure in a replicated experiment. Pesticide exposure was assessed, using pads placed inside and outside three types of protective coveralls. Potential total body accumulation rates, excluding hands, as calculated from outside pads, depended strongly upon the rate at which pesticide left the spray nozzles. When these total body rates were normalized for spray rate, the mean results, in mg-deposited/kg-sprayed, ranged from 166 to 1126, depending upon the compound applied and the application device. Overall penetration of pesticide through a disposable synthetic coverall was 3 +/- 1% for chlorpyrifos and fluvalinate, and 35 +/- 9% for ethazol. Penetration through a reusable treated twill coverall was 19 +/- 6% for chlorpyrifos, 22 +/- 13% for fluvalinate, and 38 +/- 5% for ethazol. PMID- 8239719 TI - Personal exposure to nitrogen dioxide from indoor heaters and cooking stoves. AB - The personal exposure to NO2 generated from various heaters and cooking stoves were studied, using 85 university students. The students attached NO2 filter badges to their chests or collars and wrote down the period of time for heating and cooking for 1 week. Types of heaters and smoking habits were described through a questionnaire. The urinary hydroxyproline/creatinine ratio (HOP/C) was examined as a biomarker for health effects. The outdoor NO2 concentration during the study period was 13.5-13.7 micrograms/m3. Smoking and the usage of electric heaters did not affect the exposure to NO2. Exposure increased according to the length of time kerosene heaters or oil fan heaters were used. The NO2 concentration during the heating by a kerosene heater and an oil fan heater was calculated to be 219 and 474 micrograms/m3, respectively. The correlation between the period of cooking and personal exposure was also observed. The NO2 levels during cooking were calculated to be 290 micrograms/m3. Using these calculated values of NO2 concentration, it is possible to presume the personal exposure levels from the length of time heaters and cooking stoves are used even if the subjects do not attach the filter badges. Neither smoking nor exposure to NO2 were associated with the increase of urinary HOP/C. PMID- 8239720 TI - Is a significant difference always significant? PMID- 8239721 TI - The client with substance abuse/mental illness: mandate for collaboration. AB - Persons who are dually diagnosed with substance abuse and mental illness present a significant challenge to mental health professionals in all phases of treatment. The authors explore some of the factors that contribute to the confusion surrounding the care of these persons, including the prevalence and complexity of the problem; current understanding of the effect of drug-induced neurophysiological changes on the central nervous system and the cognitive function/dysfunction of persons with a major mental illness; the difficulties presented by parallel treatment systems for mental illness and chemical addiction; and the need for a shift to interdisciplinary education to integrate services into the practice setting. PMID- 8239722 TI - Applying the "Model of recovering alcoholics' behavior stages and goal setting" to nursing practice. AB - In 1990, the author developed the Model of Recovering Alcoholics' Behavior Stages and Goal Setting as a result of grounded theory research. The model explains the natural progression of recovery through four behavior stages. Each stage is characterized by specific patient goals. Application of the model provides a framework for nursing care planning that is both stage-focused and patient centered. In this article, the author discusses nursing intervention in relation to the model. PMID- 8239723 TI - Tourette's syndrome, Part I: Definition and diagnosis. AB - Tourette's syndrome (TS) is a neuropsychiatric disorder characterized by a changing repertoire of motor and phonic tics that typically begin in childhood and exhibit a fluctuating course. Obsessive-compulsive symptoms and problems of inattention, overactivity, and impulsiveness are present in some cases. Historically, TS has been viewed as a severe disorder. However, data from epidemiologic and family-genetic studies indicate that it shows a wide range of severity. This article, the first of two parts, examines the current notions concerning the origin, pathophysiology, and diagnosis of TS. The second article will present three cases and describe contemporary approaches to treatment. PMID- 8239724 TI - Tourette's syndrome, Part II: Contemporary approaches to assessment and treatment. AB - Clinical assessment of a child with Tourette's syndrome (TS) includes a careful review of motor and phonic tics. In addition, commonly associated problems of such as obsessive-compulsive symptoms, or symptoms of attention deficit hyperactivity disorder (inattention, impulsiveness, and overactivity) should also be evaluated. Treatment almost always includes education of the child, family, and school personnel concerning the natural history and behavioral boundaries of the disorder. Other treatment interventions depend to a great extent on the primary source of impairment. This article, the second of two parts, presents three illustrative cases and reviews current treatment interventions for children and adolescents with TS. PMID- 8239725 TI - An attributional analysis of formal caregivers' perceptions of agitated behavior of a resident with Alzheimer's disease. AB - This study uses an attributional framework to examine the relationships between attributions, feelings, expectations, and caregiving behaviors of 49 dementia caregivers. Principal components factor analysis was used to reduce attribution, feeling, expectation, and caregiver behavior items to their basic components. A series of multiple regressions was used to analyze the data. Attributions were a significant predictor of preventable/reversible expectations, with specific causation being the strongest predictor. Attributions were also a significant predictor of expectations of limited predictability and controllability, with general causation being the explanatory variable. Findings support the importance of examination of attributions and their impacts. PMID- 8239726 TI - Relapse management in neurobiological disorders: the Moller-Murphy Symptom Management Assessment Tool. AB - The Moller-Murphy Symptom Management Assessment Tool (MM-SMAT) was developed to assist in the identification of psychiatric symptoms, health, and environmental variables, problems related to medication management and provide a common language for everyone involved in the ongoing management of neurobiological disorders to use in identifying the frequency, intensity, and duration of symptoms. The Murphy Wellness Model is a framework that provides insight into health, environment, and attitudes/behavior variables that became triggers of relapse. The combined use of this model and the MM-SMAT provides data to be used for anticipatory guidance of relapse management and the development of symptom specific interventions. PMID- 8239727 TI - Explorations of close friendship: a concept analysis. AB - Friendship is a meaningful and highly significant human activity. Every psychiatric nurse would acknowledge the value of friendship to mental health. But what is friendship? Development of theoretical understanding and research based interventions using friendship are hampered by inconsistent definitions of friendship. Using concept analysis, the author discusses the characteristics of close friendship and its distinguishing features. The definition created has potential implications for future theory development, research, and nursing care. PMID- 8239728 TI - A triage model of psychotherapeutic group intervention. AB - The cofacilitator of a veteran's therapy group conducted by a Veterans Administration (VA) clinical nurse specialist uses his participant-observer role to analyze the theoretical constructs used in the group. A unique group psychotherapy model blending nursing triage techniques with "hot seat" client focus and interpersonal theory is presented and compared with similar models. Clinical issues arising from its use among combat veterans with post-traumatic stress disorder at a New Hampshire VA Medical Center are also discussed. PMID- 8239729 TI - Letter from China (no. 3) PMID- 8239730 TI - [Urologic laparoscopic surgery]. PMID- 8239731 TI - [Laparoscopic surgery instrumentation]. AB - Today the urologist can perform many of the conventional surgical procedures by laparoscopic methods, which could not have been developed and improved without technological support. Industry has continued to provide solutions to each new requirement of laparoscopic surgery through more ergonomic materials and instruments. The different equipment and instruments for performing laparoscopic procedures are analyzed and described in the present article. PMID- 8239732 TI - [Anesthesia in urologic laparoscopic surgery]. AB - Although minimally invasive, urological laparoscopic surgery is still a major surgery and has special characteristics which should not be ignored. Our protocol includes: premedication with diazepam and atropine, preinduction with fentanyl, induction with propofol, followed by atracurium or succinylcholine for tracheal intubation. Anesthesia is sustained with continuous pump infusion of propofol at gradually lower doses and is discontinued on removing the abdominal trocars. Muscle relaxation throughout the operation is maintained with atracurium in continuous infusion and is discontinued at the same time as propofol. Intraoperative analgesia is achieved with bolus administration of fentanyl. We routinely use vesical and nasogastric catheters; the latter is removed at the end of the operation. Similarly, compressive bandaging is done for the lower limbs in all patients. Intraoperative monitoring includes ECG, heart rate, arterial blood pressure (noninvasive method), end expiratory CO2, O2 saturation, minute/volume, tidal volume and respiratory rate, airway pressures, temperature and diuresis. Pulmonary ventilation is by IPPV with a mixture of oxygen and air, maintaining FiO2 at 0.4. Nitrous oxide is not utilized, therefore the airways were only used for lung ventilation and not for the administration of inhalatory anesthetic agents. The higher increments of end expiratory CO2 of up to 48 mm Hg were observed at the end of the procedure following peritoneal desufflation. In summary, the technique of choice is total i.v. anesthesia with propofol and monitoring as complete as possible (noninvasive). Furthermore, capnographic and capnometric control of end expiratory CO2 is warranted. PMID- 8239733 TI - Laparoscopic entry and exit. AB - Minimal access surgery has emerged as an acceptable means of performing therapeutic and diagnostic surgical procedures. Although the gynecologist has utilized the laparoscope for several decades, only recently has laparoscopic surgery gained increased acceptance in the general surgical and urologic community. The number of laparoscopic procedures being performed routinely is now extensive and growing rapidly. It appears that the critical step in performing laparoscopic surgery is the establishment of pneumoperitoneum and the placement of the trocars. This initial part of the procedure when performed properly ensures safe and reliable access allowing the procedure to commence. Difficulties with this part of the procedure may preclude the operative procedure and more importantly may result in potentially severe injuries. Therefore a detailed discussion of the method of performing laparoscopic entry and exit will provide the urologist with the essentials to perform safe and successful laparoscopy. PMID- 8239734 TI - The application of laparoscopy to retroperitoneal surgery in urology. AB - The application of minimally invasive laparoscopic surgical techniques has provided the urologist with access to retroperitoneal structures previously available only through an abdominal incision. The enhanced visualization provided by video-imaging as well as the ease of access to the retroperitoneum through an incision in the posterior peritoneal envelope has facilitated the manipulation of many retroperitoneal organs. In particular these techniques have been applied to the clinical management of several urologic malignancies, such as adenocarcinoma of the prostate, transitional cell carcinoma of the bladder, and squamous cell carcinoma of the penis. A transperitoneal laparoscopic pelvic lymphadenectomy appears to provide adequate surgical staging of regional pelvic lymph nodes in these malignancies, while providing the patients with reduced morbidity. Additionally, transperitoneal access may be accomplished for an internal spermatic vein ligation, as well as approaches to the kidney, ureter and retroperitoneal lymph nodes. The surgical options are limited only by the availability of the instrumentation and the creativity of the surgeon. However, the indications for these procedures remain unclear and must await further information regarding actual benefits in the form of reduced patient morbidity and cost. The challenge for the future is to more completely define the indications and selection criteria for laparoscopic surgery. PMID- 8239735 TI - Laparoscopic nephrectomy: current status. AB - In this manuscript we describe our initial experience with 22 patients who underwent laparoscopic nephrectomy at our institution. Of the 22 patients, 16 patients had benign disease, and 6 patients had malignant disease. Of the 6 patients with malignancy, 3 patients underwent laparoscopic nephroureterectomy for transitional cell cancer of the upper urinary tract. In 21 patients, a transabdominal approach was utilized, whereas 1 patient underwent laparoscopic nephrectomy using an entirely retroperitoneal approach. Laparoscopic nephrectomy showed lengthy operative times, but resulted in reduced postoperative discomfort, shortened hospital stay, and rapid convalescence. As laparoscopic nephrectomy has been performed with reproducible success by various other groups worldwide, further refinements in surgical technique, paired with advances in laparoscopic instruments designed for stapling and suturing, will most likely result in an increasing application of the principles of laparoscopy to diseases of the kidney, renal pelvis and ureter. PMID- 8239736 TI - Laparoscopic varicocelectomy. AB - Laparoscopic procedures are useful techniques for minimally invasive surgery in Urology. Varicocelectomy is also performed laparoscopically with less postoperative pain and shorter convalescence when compared to the open retroperitoneal approach. Details of the technique of laparoscopic varicocelectomy are presented and the advantages and disadvantages of the procedures are discussed. From the variety of treatment modalities, the most effective and the least invasive technique should be selected to treat varicocele testis. PMID- 8239737 TI - [Laparoscopic lymphadenectomy]. AB - The limitations of the diagnostic imaging methods in determining lymph node involvement in pelvic tumors (prostatic cancer) and the high incidence of false negatives in the intraoperative frozen biopsies done during open lymphadenectomy have prompted us to develop laparoscopic staging lymphadenectomy since November, 1990. The different imaging and invasive diagnostic methods are briefly analyzed in the present article. The laparoscopic lymphadenectomy procedure is described in detail and a summary of our series is presented. PMID- 8239738 TI - [Laparoscopic renal surgery]. AB - Spurred by the development of laparoscopic lymphadenectomy and nephrectomy, laparoscopic surgery has gained acceptance in Urology, although not as rapidly as endourology. Laparoscopic access to the kidney is easier by the transperitoneal than by the retroperitoneal approach. Insertion of a trocar through the umbilicus for the optic and utilizing three to five accessory trocars is the standard practice. In some simple procedures two accessory trocars will suffice, four trocars and periumbilical trocar are required for nephrectomy, and an additional trocar is inserted in the hypogastrium for distal ureter release in nephroureterectomy. The procedure is performed using intravenous general anesthesia with endotracheal intubation and the patient is placed in the supine decubitus position with a 45 degrees lateral tilt or in the total lateral decubitus position (nephrectomy) with a slight anti-Trendelenburg. Exposure of the kidney is achieved by opening the posterior peritoneum along the line of Toldt and, in some cases, releasing the hepatic angle or the splenic colon. The kidney can be released partially or completely depending on the type of surgery. It is easier to perform an extended nephrectomy since dissection is performed better between the capsule of Gerota and the pararenal tissue. The renal vessels can be controlled with clips or an automatic stapling/cutting device. Laparoscopic access to the mid and upper ureter is very simple and the only difficulty encountered is in its correct identification. The current and future applications of laparoscopic renal surgery are discussed, including some original procedures that have been performed by the authors, such as in situ excision of a renal tumor and two pyelolithotomy procedures in solitary kidney. PMID- 8239739 TI - [Laparoscopic cystoplasty]. AB - An endoscopic cystoplasty technique is described for the first time herein. The foregoing was performed in a patient with a previous right nephrectomy due to genitourinary tuberculosis and a microbladder with marked dilatation of the ureter. The procedure commences with the insertion of a 10 mm trocar through the umbilicus and a 12 mm trocar through each flank at the level of the umbilicus and a 5 mm trocar is placed in each iliac fossa. The peritoneum is divided and the bladder wall is dissected free up to the pelvic floor. The ureter is then dissected and cut as low down as possible. A minilaparotomy is performed and the ureter and a loop of intestine are brought out. A segment of the intestine is isolated and continuity is reestablished. The ureter is anastomosed to the isolated intestinal segment and reinserted. The isolated intestinal segment is placed around the bladder and fixed with one suture on each side. A small incision is made in the bladder dome and a similar incision is made very close to this one in the intestinal segment. An Endo-GIA device is inserted, with the narrow portion in the intestine and the larger one in the bladder, and fired twice for each side. Finally, a Roticulator-type stapling device is inserted through the small laparotomy incision and positioned in the precise angle. The stapling device is opened, the bladder and intestinal orifices are positioned and stapled, which completes the procedure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239740 TI - [Radical cystectomy and laparoscopic ileal conduit]. AB - Herein we describe for the first time a laparoscopic cystectomy procedure and an ileal conduit that were performed in a single session in a patient with a tumor infiltrating the right wall. The procedure starts by releasing the ureters from the iliac junction up to a point close to the bladder. The peritoneum is incised superiorly at the level of the urachus and we proceed until the space of Retzius and the lateral walls are released. The vesicouterine plica is then incised and the bladder wings are dissected with the Endo-GIA. With a straight dissector, the urethra is released and cut until the bladder is completely free within the abdominal cavity. The trocar is removed from the right flank, the incision is extended up to about 4 cms and the bladder is removed. The right ureter and an ileal loop are then brought out through the incision on the right flank. A segment of intestine is isolated and intestinal continuity is reestablished using mechanical sutures. Then the ureter is implanted at one end of the isolated intestinal segment. The other end of the segment of intestine is taken to the left flank and anastomosed extracorporeally in a similar manner to the ureter of that side. The ileal conduit is positioned transversely so it is unnecessary to take the ureter to the opposite side. Finally, a stoma is created, which the patient desired done in the left side, and the procedure is completed. Although the operating time is long, the surgical insult is minimal because the McBurney type flank incisions cause little injury to the abdominal wall. PMID- 8239741 TI - [Laparoscopic transplantectomy of renal graft. Experimental model]. AB - Renal transplant nephrectomy carries a high morbidity and mortality. Conceptually, graft removal by laparoscopy is a logical less invasive alternative. We have developed an experimental model for renal autotransplantation in the pig, with laparoscopic graft removal eight weeks following autotransplantation. Although the clinical indication for laparoscopic nephrectomy of a failed graft will provoke controversy, a nonfunctioning renal graft with intolerance and no rupture or infection late post-transplantation theoretically could be the indication for this procedure. PMID- 8239742 TI - [Laparoscopic ureterolithotomy]. AB - Laparoscopic techniques have been proposed for the management of different pathologies of the urinary system. This study reports our initial experience in laparoscopic surgery as an alternative treatment of ureteral calculus. Five patients with proximal ureteral calculus were submitted to laparoscopic ureterolithotomy. The technique and its possible indications are discussed. PMID- 8239743 TI - [Laparoscopic urethropexis]. AB - Herein we describe a new technique of urethropexy which is performed laparoscopically. The procedure starts with the insertion of a cystoscope and the bladder is filled to determine its limits precisely. The peritoneum is then incised at the level of the urachus until it is opened wide and we advance into the space of Retzius until the entire anterior aspect of the bladder and the urethra are released. A minimal suprapubic transverse skin incision is made up to the fascia. An atraumatic needle with 1-0 vicryl suture is introduced through the incision and grasped with the endoscopic needle holder. A suture is placed in the neck and the needle and suture are brought out at the other end of the incision. Similar sutures, up to a total of four, are placed higher up. They are tied over the fascia and the procedure is completed. This technique has the advantage over the so-called needle techniques in that it permits placing the sutures required in the exact position under visual control. Moreover, the sutures can be tied easily, safely and securely because it is done externally. PMID- 8239744 TI - [Complications of laparoscopic surgery in urology]. AB - More and more laparoscopic procedures are performed in Urology each day. Attempts at reducing the invasiveness of conventional surgical procedures and the possibility of permitting patients to resume their activities earlier have considerably extended the indications for laparoscopy. Laparoscopic access to the urinary system is not simple since it is almost completely located in the retroperitoneal region and peritoneal insufflation with CO2 puts organs that were previously protected within the abdominal cavity at risk. The urologist must operate in a surgical field that was previously unfamiliar to him, with no stereoscopic vision and using instruments without all of the functions of those of conventional surgery. The complications of laparoscopic surgery may present intraoperatively or late postoperatively. Correct patient selection and preparation can prevent many of the complications that arise during trocar insertion. The physiological response to peritoneal insufflation with CO2 must be well understood if the possible complications arising from pneumoperitoneum are to be prevented or correctly treated. The different complications of laparoscopic surgery in Urology are described. The potential problems of the different stages of the procedure, the pathophysiological aspects, prevention and treatment are discussed. PMID- 8239745 TI - [Malignant rhabdoid tumor of the central nervous system. Apropos of a case with immunohistochemical and ultrastructural studies, and review of the literature]. AB - The authors report a new case of cerebral malignant rhabdoid tumour in an eight year old girl. The diagnosis was confirmed on immunohistochemical and ultrastructural examinations showing cytoplasmic perinuclear inclusions composed of whorled filaments of vimentin. Base on a review of six cases reported in the literature, they analyse the clinical, histological, immunohistochemical and ultrastructural features of this exceptional central nervous system neoplasm. Different histogenetic hypotheses are discussed. PMID- 8239746 TI - [Malignant teratoid medulloepithelioma of the ciliary body. Apropos of a case and review of the literature]. AB - A case of ocular medulloepithelioma is reported and the authors review the literature concerning these rare tumors. This report concerns a ten-month-old girl in whom ophthalmic examination showed a large tumor arising in the ciliary body with such rapid progression that enucleation was necessary. Pathological findings were compatible with a malignant teratoid medulloepithelioma. According to preceding descriptions, slit lamp examination and ultrasonography are the best approach to the diagnosis. Medulloepithelioma is a tumor composed of poorly differentiated neuroepithelial cells and the predominant histological feature is a net-like pattern. Teratoid forms contain cartilage or brain tissue or bundles of skeletal muscle. The main criterion for malignancy is the presence of neuroblastic cells. Local excision is sufficient when the tumor is mall, otherwise enucleation or even exenteration may be necessary. The prognosis is generally good even when the tumor presents malignant features on pathological examination, provided treatment is instituted rapidly. PMID- 8239747 TI - [Mullerian adenosarcoma. Apropos of a case]. AB - Mullerian adenosarcoma is an uncommon disease of the female genital tract with a special place among mixed mullerian tumors because of its slow and more favorable clinical course. This tumor is composed of a benign epithelial contingent and a sarcomatous one. A case of Mullerian adenosarcoma clinically characterized by an atypical outset is described and the literature is reviewed. PMID- 8239748 TI - [Inclusion of microfilaments and conventional cytology. Two complementary techniques applied to products of puncture with fine needle]. AB - A technique has been developed to complete the processing of fine needle aspiration biopsy material of deep and/or superficial organs: a method of collection and "prefixation staining" allowing visualisation of very small coherent fragments (microfragments) obtained by fine needle aspiration biopsy and which can consequently be submitted to histological examination. This method optimises CT-guided fine needle aspiration biopsies which, up until now, were only submitted to cytological examination (smears on slides, cell pellets, concentration and cytocentrifugation). Histological analysis of microfragments of 35 aspiration biopsies of various organs (pancreas, liver, adrenals, retroperitoneal nodes, paravertebral and mediastinal masses, etc.) was contributive in 71.4% of cases (25/35). In conclusion, the combination of cytological and microfragment inclusion techniques increased the diagnostic yield to 80% of cases. PMID- 8239749 TI - Fibronectin expression in salivary gland pleomorphic adenoma. AB - The distribution of fibronectin in the normal and pleomorphic adenoma of the salivary gland was investigated in forty cases of surgically resected tumor specimens. The immunoreactivity of fibronectin using polyclonal antibody (anti fibronectin A2 45, Dakopatts, Denmark) was detected in the cytoplasm of intercalated duct cells and certain striated duct cells in normal salivary glands. Pleomorphic adenoma showed few fibronectin positive cells with a scattered or isolated distribution within the epithelial tumor elements of tubulo ductal or solid foci of tumor cells. Modified myoepithelial cells and luminal cells of tubulo-ductal structures reacted strongly to fibronectin. In hyalinous, myxoid and chondroid areas, fibrillary myoepithelial cells expressed strongly positive reactivity but with diffuse staining in the myxoid and chondroid areas suggesting that fibronectin plays an important role in tumor cell-extracellular matrix interactions to produce the complex histopathological features of pleomorphic adenoma. PMID- 8239750 TI - The role of fine needle aspiration cytology in the diagnosis of primary skin tumors. AB - This study presents the results of 544 fine needle aspirations of primary skin tumors, (275 basal cell carcinomas, 186 squamous cell carcinomas, 41 melanomas, 5 Kaposi sarcomas, 1 sebaceous epithelioma, 1 Merkel tumor, 1 glomangiosarcoma and 34 benign tumors). The purpose of the study was to investigate the accuracy of fine needle aspiration cytology for the diagnosis of skin tumors and to assess its clinical value. 21-27 Gauge needles used and the specimens were stained by MGG and Pap methods. Immunoperoxidase stains were applied in cases with a problem of differential diagnosis. The histological examination of 382 of the cases revealed one false positive cytological result. Persuaded by our results we believe that fine needle aspiration can give highly reliable information concerning the histological type of primary skin tumors with an equivocal clinical examination. Furthermore, by detecting relapses of previously treated lesions, this method is particularly useful in the management of a great number of patients. PMID- 8239751 TI - [Diagnosis of hepatocellular carcinoma by needle puncture--aspiration and cytological test. The experience in Cameroon]. AB - The authors report their experience of the diagnosis of hepatocellular carcinoma using needle aspiration cytology over a four year period (1989-1992). In 116 of the 119 patients presenting with histologically proven hepatocellular carcinoma, the diagnosis was previously established by cytology. This method has a 97% sensitivity. Also, in forty patients without hepatocellular carcinoma, previous cytology did not detect any carcinomatous cells and therefore recorded a 100% specificity. Thus, cytologic diagnosis of hepatocellular carcinoma is possible. In countries with limited resources in which this carcinoma is widespread, needle aspiration cytology needs to be well known and used. PMID- 8239752 TI - [Histogenetic study of corpus luteum in the human ovary]. AB - The authors present a histological study of the formation of the corpus luteum after follicular rupture. According to the classical conception, the corpus luteum is derived from luteinisation of the granulosa cells: the theca interna participates in its formation only to a lesser degree. The authors criticize this histogenesis and describe the corpus luteum as being entirely derived from thecal cells. Large and small luteal cells are all derived from mesenchymal elements of the ovarian stroma, which are the precursors of steroid-secreting cells. According to the authors, granulosa cells derived from the primary sex cords of coelomic mesothelial origin only play a support role for the oocyte. After expulsion of the oocyte from the dominant follicle, the granulosa cells which remain in the follicular cavity disappear without a trace. PMID- 8239753 TI - [The Cordeliers University Center. A short guide for a visitor]. AB - The history of the Pierre and Marie Curie University Cordeliers Center is summarized. A few traces of the history of this part of the latin quarter still remain. The oldest souvenir is the plain but impressive Refectory of the Cordeliers. This Refectory is all that is left of the great Convent of the Cordeliers, and it is a reminder of the important position that theologians of the Order of Saint-Francis held in Paris. Marat's body was laid in state in the Convent Church, which used to be a royal burial place. In the hall stands the statue of Aesculapius as a reminder of the important medical reforms that took place here: the Ecole de Sante de Paris created by the Convention: the Hopital des Cliniques with its lecturing chairs; Dissection School; School of Practical Work of the Paris Medical Faculty. PMID- 8239754 TI - Adenomyoepithelioma of the breast. Report of two cases. AB - The clinical and pathologic findings of two cases of adenomyoepithelioma of the breast are reported. Aspiration cytology was performed in one case and was interpreted as malignant. Conversely, histological features revealed their typical architectural arrangement and immunohistochemical profiles confirmed that myoepithelial (actin and S-100 protein +) and epithelial (keratin and epithelial membrane antigen +) cells were active components in both tumours. PMID- 8239755 TI - Rheumatoid nodules: differential diagnosis and immunohistological findings. PMID- 8239756 TI - Neuropathology of the brainstem and spinal cord in end stage rheumatoid arthritis: implications for treatment. AB - OBJECTIVE: To study the detailed histopathological changes in the brainstem and spinal cord in nine patients with severe end stage rheumatoid arthritis, all with clinical myelopathy and craniocervical compression. METHODS: At necropsy the sites of bony pathology were related exactly to cord segments and histological changes, and correlated with clinical and radiological findings. RESULTS: Cranial nerve and brainstem pathology was rare. In addition to the obvious craniocervical compression, there were widespread subaxial changes in the spinal cord. Pathology was localised primarily to the dorsal white matter and there was no evidence of vasculitis or ischaemic changes. CONCLUSIONS: Myelopathy in rheumatoid arthritis is probably caused by the effects of compression, stretch, and movement, not ischaemia. The additional subaxial compression may be an important component in the clinical picture, and may explain why craniocervical decompression alone may not alleviate neurological signs. PMID- 8239757 TI - Linkage of rheumatoid arthritis with HLA. AB - OBJECTIVE: To determine whether HLA exerts a variable influence on the predisposition of siblings of probands with clinically mild and severe rheumatoid arthritis (RA). METHOD: Calculation of crude and adjusted odds ratios for concordance rates in sibships sharing two, one and no HLA haplotypes with a proband with clinically mild and severe RA, and HLA haplotype sharing in multiply affected sibships in the same clinical groups. RESULTS: Compared with a reference value of 1.0 in siblings sharing no HLA haplotypes with a proband with mild RA, siblings sharing two HLA haplotypes with a severely affected proband had a sibship concordance rate odds ratio of 9.7 (95% confidence interval 2.5 to 38.2). When adjusted for age, sex, and disease duration, the odds ratio was 7.6 (1.8 to 32.4). No other sibships showed concordance rates which were significantly higher than the reference group. HLA haplotype sharing in multiply affected sibships in which the proband had severe RA deviated significantly from random (two, one, and no HLA haplotypes shared: 53.3, 40, and 6.7%, respectively; expected 25, 50, and 25%), whereas in sibships of probands with mild RA they did not (14.6, 70.8, and 14.6%). CONCLUSIONS: In the predisposition of siblings to RA, sharing HLA haplotypes with a proband is only important if the proband has severe RA. Mild RA is not genetically linked to the HLA region. PMID- 8239758 TI - Abnormal plasma fibrinolysis in patients with rheumatoid arthritis and impaired endothelial fibrinolytic response in those complicated by vasculitis. AB - OBJECTIVES: (a) To assess plasma fibrinolytic parameters in patients with rheumatoid arthritis (RA) and to determine whether there are differences between patients with RA alone and those with RA complicated by vasculitis. (b) To determine if patients with RA respond differently to venous occlusion compared with normal subjects and to assess whether such a response differs in patients with RA alone and those with rheumatoid vasculitis. (c) To determine the extent of vascular damage in patients with rheumatoid vasculitis and if this correlates with the levels of plasma fibrinolytic parameters. METHODS: Sixty three patients with RA (38 had RA only and 25 had evidence of rheumatoid vasculitis) were recruited. Plasma levels of tissue plasminogen activator antigen (t-PA Ag), plasminogen activator inhibitor (PAI) activity, and factor VIII von Willebrand factor (vWF) were measured before and 10 minutes after venous occlusion. RESULTS: Patients with RA, with or without rheumatoid vasculitis, had higher baseline PAI levels than control subjects. The difference was statistically significant for patients with RA alone but was not statistically significant for patients with rheumatoid vasculitis. After venous occlusion, t-PA Ag levels increased significantly in normal subjects and patients with RA alone, but not in patients with rheumatoid vasculitis. Plasma levels of vWF were significantly higher in patients with rheumatoid vasculitis than in normal subjects and those with RA alone. In patients with RA alone, baseline vWF correlated positively with t-PA Ag levels, whereas a negative correlation was found between these two parameters in patients with rheumatoid vasculitis. A negative correlation between vWF and t-PA Ag levels after venous occlusion was also found in patients with rheumatoid vasculitis. CONCLUSIONS: Patients with rheumatoid vasculitis showed evidence of vascular damage with increased levels of vWF and impaired t-PA Ag release after venous occlusion, a useful measurement of endothelial reserve to remove fibrin. This may be of pathophysiological importance in the development of vasculitis in these patients. PMID- 8239759 TI - Components of health: an analysis in rheumatoid arthritis using quality of life questionnaires and clinical and laboratory variables. AB - OBJECTIVE: To gain insight into the overlap between additional information supplied by recently developed health status instruments for rheumatoid arthritis (RA) and traditional clinical and laboratory tests. METHODS: A cross sectional study of 282 outpatients with RA was made. From each patient, variables of clinical and laboratory measurements were obtained and the modified health assessment questionnaire (MHAQ) and a Dutch quality of life questionnaire, the IRGL, were completed. These variables were analysed for their interrelationship. RESULTS: Clinical and laboratory variables correlated significantly with the scales of the physical dimension and the disease impact scale of the IRGL. Their significant correlations with the IRGL psychological scales were weak. There were no significant correlations between any of the traditional variables and the IRGL social scales. Factor analysis yielded five factors: functionality, pain, depressive mood, social support, and laboratory. The laboratory factor is a measure of the disease process. The other four factors provide a health model. CONCLUSION: The results suggest that the IRGL questionnaire that was studied covers a complete health model that incorporates aspects of health that are not measured by clinical and laboratory tests. A simple questionnaire for the psychological and social dimension of health status, however, would probably be more cost effective and easier to use in clinical practice. PMID- 8239760 TI - Radiographic assessment of patellofemoral osteoarthritis. AB - OBJECTIVES: To determine the feasibility of assessing patellofemoral osteoarthritis using the 'skyline' view and to compare its reproducibility with the standard lateral view. METHODS: Fifty patients attending a rheumatology outpatient department with osteoarthritis of the knee had standard radiographs taken of both knees: standing weightbearing anteroposterior; lateral supine radiograph in 30 degrees of flexion; and a skyline view of the patellofemoral joint. After an initial training period using 20 sets of films the remaining 30 sets were read blind by five observers. Intraobserver and interobserver variability was assessed using the kappa statistic. The minimum joint space in each compartment was measured using a ruler. Two views of a single normal subject were measured to determine the effect of knee flexion. RESULTS: The final 30 study films were from 20 women and 10 men, median age 72.5, range 18-91 years. A grading system comprising assessment of osteophytosis, joint space narrowing, sclerosis, cysts, and attrition could easily be applied to the skyline patellofemoral view. Intraobserver reproducibility was better than the interobserver reproducibility for all features. The skyline view performed more reproducibly and over a wider range of categorisation for joint space narrowing than the lateral radiograph. Measurement using a ruler was easy to perform and precise to within 1 mm for the medial tibiofemoral and lateral facet of the patellofemoral joint. In normal knees the degree of flexion significantly affected the measurement. CONCLUSIONS: Radiographic grading of the skyline patellofemoral view is readily achieved, is more reproducible than assessment of the lateral view, and allows more precise localisation of change. Such views should be considered in radiological surveys of osteoarthritis of the knee. PMID- 8239761 TI - Oxidative DNA damage and cellular sensitivity to oxidative stress in human autoimmune diseases. AB - OBJECTIVES: To estimate the extent of genomic DNA damage and killing of lymphocytes by reactive oxygen intermediates in autoimmune diseases. METHODS: 8 Oxo-7-hydrodeoxyguanosine (8-oxodG), a promutagenic DNA lesion induced by reactive oxygen intermediates, was measured by high performance liquid chromatography, coupled with electrochemical detection, in hydrolysates of DNA which had been extracted from lymphocyte and polymorphonuclear leucocyte fractions of human blood. In addition, human primary blood lymphocytes stimulated by concanavalin A were assayed for cytotoxicity induced by hydrogen peroxide on day 0, by assessing cell proliferation during seven days of culture. RESULTS: Constitutive 8-oxodG was detectable (mean (2 SEM) moles 8-oxodG/10(6) moles deoxyguanosine) in DNA isolated from normal human blood lymphocytes (68 (8), n = 26) and polymorphonuclear leucocytes (118 (24), n = 24). Lymphocyte DNA from donors with the following inflammatory autoimmune diseases contained significantly higher levels of 8-oxodG than that from healthy donors: rheumatoid arthritis (98 (16)), systemic lupus erythematosus (137 (28)), vasculitis (100 (32)), and Behcet's disease (92 (19)). Lymphocyte 8-oxodG levels in non autoimmune controls and patients with scleroderma were not significantly different from those of healthy controls. The levels of 8-oxodG were significantly higher in the DNA from normal polymorphonuclear leucocytes than in paired DNA samples from normal lymphocytes, but there were no differences between levels of 8-oxodG in polymorphonuclear leucocytes from normal subjects and the patients studied. Levels of 8-oxodG did not correlate with disease duration, disease severity, or age. Lymphocytes from patients with systemic lupus erythematosus and rheumatoid arthritis, but not those with scleroderma, also showed cellular hypersensitivity to the toxic effects of hydrogen peroxide. CONCLUSION: There was increased genomic DNA damage, and increased susceptibility to cytotoxic killing by hydrogen peroxide, in lymphocytes from patients with certain autoimmune diseases. These results might be explained by defective repair of DNA damage or by increased production of reactive oxygen intermediates in inflammation. Although more direct studies are needed, the evidence available favours the former explanation. PMID- 8239762 TI - Role of the CS1 adhesion motif of fibronectin in T cell adhesion to synovial membrane and peripheral lymph node endothelium. AB - OBJECTIVES: It has previously been shown that the very late antigen-4/vascular cell adhesion molecule-1 (VLA-4/VCAM-1) pathway functions as a receptor/ligand interaction system mediating the recruitment of activated lymphocytes to inflamed synovium of patients with rheumatoid arthritis. This study was performed to determine whether VLA-4 also affects lymphocyte adhesion to inflamed synovium through interaction with the alternatively spliced CS1 domain of fibronectin. METHODS: The effect of the synthetic peptide CS1 on lymphocyte binding to human synovial and peripheral lymph node high endothelial venules (HEVs) was measured in an in vitro frozen section assay. RESULTS: In the presence of the CS1 peptide or antibody to fibronectin, significant inhibition of binding was observed (54 and 51% respectively). Blocking with antibody to VCAM-1 yielded inhibition of binding to 46% of the control value. Maximum inhibition of binding was obtained with a combination of antibody to VCAM-1 and CS1 (65%) and with antibody to VLA-4 alpha (68%). Blocking the classical fibronectin receptor with antibody to VLA-5 alpha gave a slightly lower inhibition at 42%. In normal peripheral lymph nodes, the synthetic peptide CS1 and antibodies to fibronectin and VLA-5 also partially inhibited T cell binding to HEVs (45, 47, and 52% respectively). CONCLUSION: These results show that fibronectin mediates lymphocyte-HEV interactions not only through its classical VLA-5 receptor, but also through its CS1 adhesion motif in inflamed synovium and peripheral lymph nodes. PMID- 8239763 TI - Presence of foam cells containing oxidised low density lipoprotein in the synovial membrane from patients with rheumatoid arthritis. AB - OBJECTIVE: Increased concentrations of lipid peroxidation products have been described in the serum and synovial fluid from patients with rheumatoid arthritis. A large proportion of the unsaturated lipids in human extracellular fluids is a component of low density lipoprotein (LDL). The oxidative modification of LDL, and its subsequent uptake by macrophages, has been implicated in the pathogenesis of atherosclerosis, but not of rheumatoid arthritis. This study aimed to assess whether oxidatively modified LDL was present in the rheumatoid synovium. METHODS: A polyclonal antiserum raised in rabbits against oxidised LDL (o-LDL) was used to perform an immunohistochemical study of a series of synovial biopsy specimens from patients with rheumatoid arthritis. RESULTS: Collections of positively stained macrophages, arranged in a linear fashion and with the morphological characteristics of foam cells--that is, 'fatty streaks', were identified around blood vessels within the intimal connective tissue. In addition, scattered, positively stained foam cells were present in association with deposits of fibrin. These staining patterns were absent from control synovial membranes (traumatic knee injuries). CONCLUSIONS: The findings in all rheumatoid patients studied suggest that atherosclerosis and rheumatoid arthritis have analogous pathogenetic features. PMID- 8239764 TI - Stress fractures of the pubic rami in rheumatoid arthritis. AB - OBJECTIVE: To draw attention to and detail the features of stress fractures of the pubic rami in patients with rheumatoid arthritis (RA). METHODS: Twenty two cases were collected prospectively over a four year period from patients undergoing active rheumatological surveillance in Leeds. Both old and new fractures were included. Information was obtained from the patients, clinical notes, and radiological investigations. RESULTS: All patients were women (mean age 68.1 years) with longstanding (mean disease duration 24.2 years) seropositive RA. Nineteen (86%) were receiving regular prednisolone treatment and all patients had radiological evidence of osteoporosis, with vertebral crush fractures in 10 (63%) of the 16 who had vertebral x ray examination. There was no biochemical evidence of osteomalacia. Nineteen (86%) presented with pain in the low back, groin, or hip and three were asymptomatic. Pain developed gradually in seven with an acute onset in the remainder. Six gave a history of a fall but only seven were x rayed at the onset of symptoms and initial radiographs were negative in five of these. In eighteen the fracture had either minimal or no effect on their mobility. Fractures affected all four pubic rami and in four all four were fractured. All but one patient (who had multiple fractures) made an uneventful recovery over two to four weeks with conservative management. CONCLUSIONS: Stress fractures of the pubic rami in RA appear to be more common than had been recognised. The low grade nature of symptoms, the minimal effect on mobility, and the absence of significant trauma are typical features and suggest that many more stress fractures may go unrecognised. PMID- 8239765 TI - Benign rheumatoid nodules in a woman with chronic lymphocytic leukaemia and borderline lepromatous leprosy. AB - OBJECTIVES: To report benign rheumatoid nodules in a woman with chronic lymphocytic leukaemia and borderline lepromatous leprosy and to summarise the features of the patients with adult onset benign rheumatoid nodules. METHODS: A 66 year old woman with chronic lymphocytic leukaemia and borderline lepromatous leprosy who presented with subcutaneous elbow nodules, which were at first suspected to represent either progression of her haematological disease or leprosy, is described. The clinical characteristics of our patient and previous reports of another 24 subjects with adult onset benign rheumatoid nodules are reviewed. RESULTS: Biopsy of the patient's subcutaneous lesion disclosed the histopathology of a rheumatoid nodule; serological and clinical evaluations for rheumatoid arthritis and other rheumatoid nodule associated systemic diseases were negative. Adult onset benign rheumatoid nodules are clinically and histologically identical to those found in patients with rheumatoid arthritis. They often appeared in women during their 20s, frequently resolved spontaneously or were adequately treated by excision, and recurred in about one third of patients. The lesions were located in the ocular adnexa in 60% of patients. The most common lesional sites in patients with non-ocular benign rheumatoid nodules were the elbows, feet, and knees. None of these patients subsequently developed rheumatoid arthritis or other rheumatoid nodule associated diseases during follow up periods of as long as 20 years. CONCLUSION: The appearance of subcutaneous nodules is often the harbinger of an associated systemic disorder. Although benign rheumatoid nodules occur infrequently in adults, they should be considered in the differential diagnosis of new nodular lesions. PMID- 8239766 TI - Patients with antiphospholipid antibodies and venous thrombosis should receive long term anticoagulant treatment. AB - OBJECTIVE: To determine whether the finding of antiphospholipid antibodies in patients with venous thromboembolic episodes should influence the duration of treatment with anticoagulant drugs by mouth. METHODS: A retrospective study was carried out in 19 patients with antiphospholipid antibodies and a history of venous thromboembolic episodes. The median follow up from the first venous thromboembolic episode was 93 months and the median age at this episode was 26 years. The patients had in total 34 venous thromboembolic episodes. The total follow up period comprised 32 periods with and 23 periods without anticoagulant drugs. RESULTS: The probability of being free of recurrent venous thromboembolic episodes, calculated by the Kaplan-Meier method, was significantly influenced by the use of anticoagulant drugs. Patients receiving oral anticoagulants had at eight years a 100% probability of survival without recurrence, whereas patients in whom anticoagulant drugs were stopped had a 50% probability of a recurrent venous thromboembolic episode at two years, and a 78% probability of recurrence at eight years. CONCLUSION: Patients with venous thromboembolic episodes and antiphospholipid antibodies have a high risk for recurrent venous thromboembolic episodes and long term treatment with anticoagulant drugs by mouth is an effective prophylaxis. PMID- 8239767 TI - Towards a coherent terminology of gout. PMID- 8239768 TI - Enterobacter cloacae vertebral infection in a heroin addict with HIV infection. PMID- 8239769 TI - Reinfection in paediatric Lyme borreliosis. PMID- 8239770 TI - Health screening: the temporomandibular joint. PMID- 8239771 TI - A current view of gastric cancer in the US. AB - In the US, the remarkable decline in the incidence of gastric cancer during the mid-portion of this century has leveled off during the last two decades as an equally remarkable and poorly understood increase in the percentage of the generally more unfavorable cardia cancers has become apparent. The importance of H. pylori infection is being actively investigated and treatment to reduce the infection may offer a means of decreasing the disease, particularly in areas of high incidence. The potential danger of inciting gastric cancer by the prolonged use of drugs that severely reduce or eliminate gastric acid has been mentioned, but the degree of risk must await the passage of years before it can be properly evaluated. "Early gastric cancer" or, probably more appropriately, "superficial gastric adenocarcinoma" continues to comprise a relatively small segment of gastric cancers in the US and most Western countries. Seventeen per cent of cases in the ACS series were classified as stage I, a much higher incidence than reported for early gastric cancer in most individual North American series. The ACS report suggests "special education of the surgeon in the requisites for adequate gastrectomy with node dissection, coupled with effective adjuvant therapy" as a means of improving results in the US. This is a significant consideration because, unfortunately, gastric surgery for ulcer or cancer no longer plays the important role it did in past decades in many US surgical training programs. As has been demonstrated in Japan and in certain larger US series, excellent surgical technique, particularly for cardia tumors, plays an important role in obtaining improved results. The value of radical lymph node dissection continues to be controversial in US cases, and a successful chemotherapeutic regimen has yet to be found. Subtotal gastric resection, as noted in the ACS report, continues to be the procedure of choice in the US for most gastric cancers, even for cardia cancers. Although there is no improvement in survival, quality of life is thought by some to be better after total gastrectomy for cardia cancers rather than proximal subtotal esophagogastrectomy. However, equally important for improved survival is the ACS recommendation of earlier referral for gastric surgery patients with precursor lesions, but the lack of improvement in the pathological stage of disease in the two ACS time periods suggests that little progress is being made in this country in this regard. PMID- 8239772 TI - Cancer of the stomach. A patient care study by the American College of Surgeons. AB - OBJECTIVE: The major purpose of this study was to document the modes of presentation, diagnostic methods, clinical management, and outcome of gastric cancer as reported by tumor registries of US hospitals and cancer programs approved by the American College of Surgeons. SUMMARY BACKGROUND DATA: Gastric cancer continues to diminish in the US, but the stage of disease and survival outcome after surgical resection is unchanged despite increased availability and sophistication of diagnostic techniques. This is in contrast to the marked improvement in survival outcome in Japanese and other Eastern series over the last decades. Possible reasons for the improved Japanese results have been earlier detection secondary to active diagnostic surveillance of the population and widespread adoption of aggressive surgical resection emphasizing wide-field node (R2) dissection. Although selected US centers using the Japanese approach report better survival data, the approach has not been widely adapted by US treatment centers. METHODS: Tumor registries at American College of Surgeons (ACS) approved hospitals were mailed a study protocol in 1987. They were instructed to review 25 consecutive patients with gastric cancer treated in 1982 (long-term study) and 25 patients treated in 1987 (short-term study). A detailed protocol included significant history, diagnostic results, staging, pathology findings, and treatment results. The data forms on 18,365 patients were returned and analyzed (11,264 patients in the long-term study and 7101 patients in the short-term study). RESULTS: Of 18,365 patients, 63% were males. The median ages were 68.4 years in males and 71.9 years in females. There was a history of gastric ulcer in 25.5% of the patients. Lesion location was upper third in 31%, middle third in 14%, distal third in 26%, and entire stomach in 10% of patients (and the site was unknown in 19%). Gastric resection was performed for 80% of upper third cancers and 85% of distal third cancers; 50% of patients with total gastric involvement had gastric resection. The extent of gastric resection varied according to location. For lower third lesions, subtotal gastrectomy was done in 55% of the cases, extended resection in 21%, and total gastrectomy in 6%. For proximal lesions, 29% had subtotal, 4.6% had total, and 41% had extended gastrectomies (including esophagus), and 13.6% had dissection of celiac nodes. The operative mortality rate was 7.2%. Staging (American Joint Committee on Cancer [AJCC]) was as follows: I, 17%; II, 17%; III, 36%; and IV, 31%. The overall survival rate reflecting deaths from all causes was 14% among 10,891 patients diagnosed in 1982, and it was 19% in patients having resection. The disease specific survival rate was 26%. The survival rate after resection was 19% and 21% for lower and mid third cancers, 10% for upper third cancers, and 4% if the entire stomach was involved. The stage-related survival rates were 50% (stage I), 29% (stage II), 13% (stage III), and 3% (stage IV). Among patients with pathologically clear margins, the survival rate was 35% versus 13% in those with microscopically involved margins, and it was 3% in those with grossly involved margins. CONCLUSION: This report of gastric cancer treatment by American College of Surgeons approved institutions in the US provides an overview of the disease as commonly treated throughout the US. Although the results are less favorable than those reported by centers with large institutional experiences with this disease and are inferior to those of the Japanese and other Eastern centers, they suggest potential for increasing survival by upstaging through earlier diagnosis and using resectional techniques demonstrated to more adequately control local regional disease. PMID- 8239773 TI - The vascular endothelium. A new horizon. PMID- 8239774 TI - Hepatic resection for cystic lesions of the liver. AB - OBJECTIVE: The purpose of this study was to report the authors' experience with hepatic resection for cystic lesions of the liver. SUMMARY BACKGROUND DATA: Past experience with aspiration, sclerosing therapy, internal drainage, fenestration, and marsupialization are of limited value. Hepatic resection has evolved into a safe operation over the last two decades. METHODS: A retrospective study of 44 patients with various cystic lesions of the liver (polycystic disease, 2; solitary or multiple congenital cysts, 19; biliary cystadenoma, 6; cystadenocarcinoma, 3; squamous cell carcinoma, 3; Caroli's disease, 5; and hydatid cyst, 6) was performed. RESULTS: After 7 trisegmentectomies, 24 lobectomies, 6 left lateral segmentectomies, and 7 nonanatomical hepatic resections, only 1 operative death occurred in a Jehovah's Witness. Symptomatic relief was complete and permanent in all of the patients with benign congenital or parasitic hepatic cysts, except for the two patients with polycystic disease of the liver. One of the 3 patients with adenocarcinoma and 3 patients with squamous cell carcinoma of the cyst wall died of tumor recurrence between 3 and 14 months after hepatic resection. CONCLUSIONS: Hepatic resection is safe and effective for cystic lesions of the liver. Symptomatic relief is complete and permanent after hepatic resection, except in cases of diffuse polycystic disease of the liver. Liver transplantation should be considered for diffuse polycystic disease of the liver when the symptoms are extremely severe. PMID- 8239775 TI - The Hepp-Couinaud approach to strictures of the bile ducts. I. Injuries, choledochal cysts, and pancreatitis. AB - OBJECTIVE: The results of operative repair of benign strictures of the bile duct after cholecystectomy, right hemihepatectomy, vagotomy and antrectomy, choledochal cysts in adults, and chronic pancreatitis, with particular reference to the use of the Hepp technique for hilar strictures and without the use of transanastomotic tubal stenting, were analyzed in 44 patients. SUMMARY BACKGROUND DATA: End-to-side bilio-enteric anastomoses have been reported to be associated with restricturing and reoperation in 12% to 25% of cases and operative morbidity and mortality rates of 10% and 5% to 8%, respectively. Long-term transanastomotic tubal stenting is widely practiced in an attempt to prevent or diminish anastomotic stricturing. METHODS: The Hepp technique of wide, accurate, mucosa-to mucosa anastomosis between the left hepatic duct and a jejunal Roux loop was used in 28 patients with hilar bile duct strictures. The same technical principle of wide side-to-side anastomosis was used in most of the lower strictures. Patients have been observed for 1 to 14 years (median, 7 years). RESULTS: The operative mortality rate was 7% (3 patients), but only 2.4% (1 patient) in 41 noncirrhotic patients. Two patients who had undergone standard end-to-side hepaticojejunostomy required reoperation (Hepp procedures) for recurrent strictures. No recurrent strictures occurred with the use of the Hepp technique for hilar strictures or wide side-to-side anastomosis for lower strictures. None of these patients experienced episodes of ascending cholangitis. CONCLUSIONS: The Hepp approach provides a safe, durable, and highly effective solution to the problem of strictures of the bile duct, including hilar strictures. Transanastomotic tube stenting is not necessary. PMID- 8239776 TI - The current role of U tubes for benign and malignant biliary obstruction. AB - OBJECTIVE: The recent experience with U tubes at Rush-Presbyterian-St. Lukes Medical Center was reviewed in order to assess their current role in hepatobiliary surgery. SUMMARY BACKGROUND DATA: Transhepatic intubation by a variety of methods has been used routinely for biliary decompression and inhibition of anastomotic stricture since the 1960s. U tubes were popularized in the early 1970s. However, little has been written about their use and efficacy in recent years. Because of the apparent benefits associated with the use of U tubes versus other stenting techniques, the authors performed this study. METHODS: The hospital and office charts of all patients who had U tubes placed between 1980 and 1992 were reviewed retrospectively. Between 1980 and 1992, U tubes were placed intraoperatively in 54 patients for biliary decompression and/or stenting. Twelve patients were operated on for benign causes of obstruction. Forty-two patients with malignant tumors underwent surgery for U tube placement in conjunction with or without tumor resection and anastomotic bypass. RESULTS: There was a 0% operative mortality rate in the benign group. In six patients, the U tube played a major role in the long-term management of their disease processes. None of these patients has had restricture since removal of the tube. In the malignant group, the 30-day operative mortality rate was 12%. After 3 months, marked clinical improvement and complete biliary decompression were achieved, with mean bilirubin levels dropping from 14.0 mg/dL to 1.3 mg/dL. No patients in the malignant group required reoperation for recurrent biliary obstruction after U tube placement. CONCLUSIONS: The use of U tubes is advocated for biliary decompression and/or anastomotic stenting in patients with benign stricture or resectable malignancy and in patients with nonresectable, malignant biliary obstruction for adequate palliation of intractable jaundice. PMID- 8239777 TI - Laparoscopic cholecystectomy for acute inflammation of the gallbladder. AB - OBJECTIVE: The aim of this study was to prospectively assess the results of laparoscopic cholecystectomy in patients with acute inflammation of the gallbladder. SUMMARY BACKGROUND DATA: Laparoscopic cholecystectomy has become the standard treatment for symptomatic gallbladder disease. Its role in the surgical treatment of acute cholecystitis has not been defined, although a number of recent reports suggest that there should be few contraindications to an initial laparoscopic approach. METHODS: All patients presenting with symptomatic cholelithiasis from October 1990 until June 1992 were evaluated at laparoscopy with intention of proceeding to a laparoscopic cholecystectomy. The gross appearance of the gallbladder was categorized as acute inflammation, chronic inflammation, or no inflammation. Ninety-eight (23.4%) of 418 patients had acute inflammation of the gallbladder: 55 were edematous, 10 were gangrenous, 15 had a mucocele, and 18 had an empyema. RESULTS: The authors assessed outcome in these patients. The frequency of conversion to an open operation was 33.7% for acute inflammation, 21.7% for chronic inflammation (p < 0.05), and 4% for no inflammation (p < 0.001). The conversion rate was highest for empyema (83.3%) and gangrenous cholecystitis (50%), while the conversion rate for edematous cholecystitis was 21.8% and for acute inflammation with a mucocele it was 7%. The median operation time for successful laparoscopic cholecystectomy for acute inflammation was 105 minutes, which was longer than that with no inflammation (90 minutes). However, the incidence of complications was not different from that for chronic or no inflammation. The median postoperative stay for patients with acute gallbladder inflammation was 2 days for successful laparoscopic cholecystectomy and 7 days for patients converted to an open operation. CONCLUSIONS: Laparoscopic cholecystectomy for acute inflammation of the gallbladder is safe and is associated with a significantly shorter postoperative stay compared to open surgery. A greater number of patients required conversion to open operation compared to those with no obvious inflammation. Conversion to open operation was most frequent for empyema and gangrenous cholecystitis, suggesting that once this diagnosis is made, excessive time should not be spent in laparoscopic trial dissection before converting to an open operation. PMID- 8239778 TI - Intraoperative study on the relationship between the lower esophageal sphincter pressure and the muscular components of the gastro-esophageal junction in achalasic patients. AB - OBJECTIVE: The lower esophageal sphincter (LES) resting tone originates from the tension of the muscular fibers of the gastro-esophageal (GE) junction. This study determined which of the muscular structures' of the GE junction are actually responsible and to what degree for the LES resting tone in achalasic patients. SUMMARY BACKGROUND DATA: Controversy still exists as to the length of myotomy on the esophageal and gastric sides of the GE junction. Experimental and clinical studies have supposed that the anatomical complex formed by the U and the sling fibers of the lesser curvature of the stomach can be part of the LES. METHODS: The variations induced on the LES resting tone by the separate division of the esophageal and gastric muscular fibers of the GE junction were studied by means of intraoperative manometry in 32 patients who underwent myotomy for achalasia. RESULTS: After surgical preparation of the GE junction, the mean pressure was 29.3 +/- 13 mmHg. After esophageal side myotomy, the mean LES pressure decreased to 13.6 +/- 7.9 mmHg (paired t test, p < 0.0005). The residual pressure was further reduced after gastric side myotomy (3.4 +/- 1.9 mmHg; paired t test, p < 0.0005). CONCLUSIONS: In achalasic patients, 45% of the LES resting tone is maintained by the gastric side anatomical component of the GE junction. The range of variability of the gastric component of the LES is wide. This information should be taken into account when performing extramucosal myotomy as therapy for esophageal achalasia. PMID- 8239779 TI - Reoperative bariatric surgery. Lessons learned to improve patient selection and results. AB - OBJECTIVE: The purpose of this study was to determine the spectrum of presentation, safety, and efficacy of operative bariatric surgery. SUMMARY BACKGROUND DATA: The only lasting therapy for medically complicated clinically severe obesity is bariatric surgery. Several operative approaches have resulted in disappointing long-term weight loss or an unacceptable incidence of complications that require revisionary surgery. METHODS: Sixty-one consecutive patients who underwent reoperative bariatric surgery from 1985 to 1990 were observed prospectively. One, two, or three previous bariatric procedures had been performed in 77%, 18%, and 5% of patients, respectively. Reoperation was required for unsatisfactory weight loss after gastroplasty or gastric bypass (61%), metabolic complications of jejunoileal bypass (23%), or other complications (16%), including stomal obstruction, alkaline- or acid-reflux esophagitis, and anastomotic ulcer. Revisionary procedures included conversion to vertical banded gastroplasty (33% of operations) and vertical Roux-en-Y gastric bypass (52% of operations); partial pancreato-biliary bypass was used selectively in four patients with severe, medically complicated obesity. RESULTS: A single patient died postoperatively of a pulmonary embolus; serious morbidity occurred in 11%. Weight loss (mean +/- SEM) after reoperation for unsuccessful weight loss was greater with gastric bypass than with vertical banded gastroplasty (54 +/- 6% versus 24 +/- 6% of excess body weight). Metabolic complications of jejunoileal bypass were corrected, but 67% of the patients were dissatisfied with their postoperative lifestyle because of changes in eating habits or weight gain (64% of patients). Stomal complications and esophageal reflux symptoms were reversed in all patients. CONCLUSIONS: Reoperative bariatric surgery in selected patients is safe and effective for unsatisfactory weight loss or for complications of previous bariatric procedures. Conversion to gastric bypass provides more effective weight loss than vertical banded gastroplasty. PMID- 8239780 TI - Small bowel obstruction after nephrectomy for Wilms' tumor. A report of the National Wilms' Tumor Study-3. AB - OBJECTIVE: This study was undertaken to define the incidence and etiology of small bowel obstruction (SBO) after nephrectomy for Wilms' tumor. SUMMARY BACKGROUND DATA: Intestinal obstruction is one of the most common postoperative complications after nephrectomy for nephroblastoma. However, few reports have evaluated risk factors for SBO. Radiation therapy has been associated with increased intestinal complications in some adult cancer patients, but this has not been reported in children undergoing cancer surgery. METHODS: Postoperative SBO occurred in 131 of 1,910 children (6.9%) enrolled in the Third National Wilms' Tumor Study (NWTS). The etiology of the SBO was bowel adhesions in 104 cases, intussusception in 17, internal hernia in 2, and uncertain in the remaining 8 children. RESULTS: The factors found to be of potential importance in explaining the incidence of SBO were higher local tumor stage, extrarenal intravascular involvement, and en bloc resection of other organs at the time of nephrectomy. The incidence of postoperative SBO was not increased in children who received postoperative radiation therapy. CONCLUSIONS: Although the overall incidence of SBO after nephrectomy for Wilms' tumor is comparable to that after other major abdominal operations in children, it can be responsible for significant morbidity. There were 4 children among the 1,910 patients with infectious complications of SBO, which contributed to their death. PMID- 8239781 TI - Incidence, diagnosis, and treatment of enteric and colorectal fistulae in patients with Crohn's disease. AB - OBJECTIVE: The authors review their experience, evaluating the incidence and examining the various modalities employed in the diagnosis and treatment of patients with Crohn's disease complicated by fistulae. SUMMARY BACKGROUND DATA: Although common, internal and external fistulae in Crohn's disease may pose challenging problems to the surgeon. METHODS: Of 639 patients who underwent surgical treatment at the University of Chicago between 1970 and 1988 for complications of Crohn's disease, 222 patients (34.7%) were found to have 290 intra-abdominal fistulae. RESULTS: A fistula was diagnosed preoperatively in 154 patients (69.4%), intraoperatively in 60 (27%), and only after examination of the specimen in 8 (3.6%). The fistula represented the primary or single indication for surgical treatment in 14 patients (6.3%) and one of several indications in the remaining patients. Of 165 patients with an abdominal mass or abscess, 69 (41.8%) had a fistula. All patients underwent resection of the diseased intestinal segment; 160 (73.1%) with primary anastomosis and the remaining 62 with a temporary or permanent stoma. The fistula was directly responsible for a stoma in only 16 patients (7.2%) and was never responsible for a permanent stoma. Resection of the diseased bowel achieved en bloc removal of the fistula in 145 cases. Removal of 93 additional fistulae required resection of the diseased bowel segment along with closure of a fistulous opening on the stomach or duodenum (n = 14), bladder (n = 35), or rectosigmoid (n = 44). When the fistula drained through a vaginal cuff (n = 4), the opening was left to close by secondary intention; when the fistula opened through the abdominal wall (n = 46), the fistulous tract was debrided. In the remaining two entero-salpingeal fistulae, en bloc resection of the involved salpinx accomplished complete removal of the fistula. There was a dehiscence of one duodenal and one bladder repair; 14 patients (6%) experienced postoperative septic complications and one patient died. CONCLUSIONS: Fistulae are diagnosed preoperatively in 69% of cases and can be suspected in as many as 42% of patients with an abdominal mass. Fistulae are the primary or single indication for surgical treatment and are directly responsible for a stoma only in a few patients. Treatment, based on resection of the diseased bowel and extirpation of the fistula, can be accomplished with minimal morbidity and mortality. PMID- 8239782 TI - Breast cancer in the elderly. AB - OBJECTIVE/METHODS: To determine the clinical behavior and outcome of breast cancer in the elderly, a series of 184 women older than age 69 years who received treatment for locoregional breast cancer at The University of Texas M. D. Anderson Cancer Center between 1976 and 1985 were studied for a median of 80 months. RESULTS: The results indicate that elderly women can tolerate standard surgical therapy and survive disease-free for many years; the breast cancer specific survival rate of patients in this study was 79% at 7 years. Although 33% of patients had stage I disease, only 10% underwent breast conservation surgery. Despite 46% of patients having stage II and 21% having stage III breast cancer, fewer than 13% received systemic adjuvant therapy. Noncompliance with breast screening guidelines was evident in that only 3% of patients had tumors detected by routine screening mammograms and only 12% by routine physical examinations. CONCLUSIONS: Women with breast cancer should be informed of treatment options and the advantages and disadvantages of each choice based on physiologic rather than chronologic age. PMID- 8239783 TI - Upper airway compromise after inhalation injury. Complex strictures of the larynx and trachea and their management. AB - OBJECTIVE: Strictures of the upper airway caused by burns have features distinct from other benign stenoses. The authors reviewed their experience with burn related stenoses to define the principles of treatment. SUMMARY BACKGROUND DATA: The combined effects of inhaled gases and heat in burn victims produce an intense, often transmural, inflammation of the airway, further complicated by intubation. The incidence of laryngotracheal strictures in survivors of inhalation injury is high, but the reported experience with their treatment is limited and often unduly separated into injuries of larynx and trachea. METHODS: Presentation, treatment, and long-term follow-up are reviewed in 9 women and 9 men age 9 to 63 years, who were evaluated over a 22 year period for chronic airway compromise after inhalation injury. There were 18 tracheal stenoses, 14 subglottic strictures, and 2 main bronchial stenoses. Laryngotracheal strictures stenosis. T-tubes were placed in 15 patients, in low subglottic or tracheal stenosis below the vocal cords, in high subglottic stenosis through the vocal cords, and as a stent after resection of subglottic stenosis. RESULTS: There were two deaths during follow-up, one from respiratory failure and one from an unrelated cause. Two patients underwent evaluation only. Early in this series, one tracheal and one laryngotracheal resection resulted in prompt restenosis. Of the remaining 14 patients, 9 are without airway support from 2 to 20 years later. Four have permanent tracheal tubes. One patient required tracheostomy 8 years after successful subglottic reconstruction. CONCLUSIONS: Strictures of the upper airway related to inhalation injury are associated with prolonged inflammation and involve larynx and trachea in a majority of patients. These complex injuries respond to prolonged tracheal stenting (mean, 28 months) and resection or stenting of subglottic stenoses with recovery of a functional airway and voice in most patients. Early tracheal resection should be avoided. PMID- 8239784 TI - Acute response of human muscle protein to catabolic hormones. AB - OBJECTIVE: The purpose of this study was to determine the acute in vivo response of human muscle protein to stress. SUMMARY BACKGROUND DATA: Prior animal and human in vitro studies have suggested that physiologic stress increases muscle protein turnover. In contrast, recent publications using a polyribosomal methodology have demonstrated a reduction in human muscle protein synthesis in vivo after surgery. METHODS: Five healthy volunteers were given a stable isotopic infusion of 1,2(13)C leucine that allowed for determination of the fractional rate of muscle protein synthesis by measuring the rate of incorporation of 13C label into vastus lateralis muscle biopsies. Simultaneous infusion of 15N lysine and quantitation of leg blood flow by indocyanine green dye dilution allowed for estimation of leg muscle protein breakdown rate (Lys Ra) and synthesis rate (Lys Rd). These measurements were performed before and then at the conclusion of a 4 hour femoral arterial infusion of the catabolic hormones epinephrine, cortisol, and glucagon. RESULTS: The catabolic hormone infusion elicited a significant (65%) increase in the leg muscle protein breakdown rate and a significant but less marked increase in the rate of muscle protein synthesis, as assessed by both an increase in the fractional rate of muscle protein synthesis of 48.5% and in lysine uptake within the leg of 32%. CONCLUSIONS: This study conclusively demonstrates that a hormonally induced stress results in a net catabolism of human muscle protein by increasing the rate of protein breakdown in excess of an increased protein synthetic rate. PMID- 8239785 TI - Laparoscopic versus conventional appendectomy. AB - OBJECTIVE: The goal of this study was to prospectively define the impact of laparoscopy on the management of patients with a presumed diagnosis of appendicitis. SUMMARY BACKGROUND DATA: While the role of laparoscopy in the management of cholelithiasis is well established, its impact on the management of acute appendicitis needs to be objectively defined and compared to that of conventional management. Several authors have predicted that laparoscopic appendectomy will become the preferred treatment for appendicitis. METHODS: Two groups of consecutive patients with similar clinical characteristics of acute appendicitis were compared. Data on the laparoscopic group were compiled prospectively on standardized forms; data on the conventional group were collected retrospectively. Operative time, hospital stay, analgesia, cost, and return to normal activities were noted. RESULTS: Seventeen consecutive patients who underwent appendectomy were compared to 18 consecutive patients who underwent laparoscopy (16 of these 18 had laparoscopic appendectomy). There was no significant difference between the two groups in terms of clinical characteristics and appendiceal histopathology. The mean operative times were 61 +/- 4.1 minutes and 46 +/- 2.9 minutes for the laparoscopy and conventional groups, respectively (p < 0.01). Hospital stay was significantly shorter in the laparoscopic appendectomy group, with 81% of patients being discharged on their first postoperative day (p < 0.001). The laparoscopic appendectomy patients required significantly less narcotic analgesia (p < 0.02). Return to normal activity was not significantly different between the two groups. The average total cost of laparoscopic appendectomy was 30% greater than that of conventional appendectomy. CONCLUSIONS: Laparoscopy is a useful adjunct to the management of patients with a presumed clinical diagnosis of acute appendicitis. PMID- 8239786 TI - Multiple system organ failure. PMID- 8239787 TI - Computerized identification of duodenogastric reflux using 24-hour gastric pH monitoring. PMID- 8239788 TI - The increasing trend toward conservative surgery for malignant melanoma. PMID- 8239789 TI - Surgical margins around primary cutaneous melanomas. PMID- 8239790 TI - Warm versus cold blood cardioplegia: a self-imposed and counterproductive dilemma. PMID- 8239791 TI - Why not close a small ventricular septal defect? PMID- 8239792 TI - A reason to close ventricular septal defect? PMID- 8239793 TI - Restrictive ventricular septal defect: how small is too small to close? AB - Historically, indications for ventricular septal defect closure have included congestive heart failure, pulmonary hypertension, aortic insufficiency with or without aortic valve prolapse, and prior bacterial endocarditis. However, controversy exists as to how the lifetime risk of an isolated, nonoperated restrictive ventricular septal defect compares with the risk of surgical closure in an asymptomatic child. Between 1980 and 1991, cardiac catheterization and elective ventricular septal defect closure (age > 1 year, pulmonary to systemic flow ratio < 2.0) were performed in 141 patients aged 1 to 23 years (mean age, 6.1 +/- 4.7 years). Mean systolic pulmonary artery pressure was 26.9 +/- 13.0 mm Hg, and mean pulmonary to systemic flow ratio was 1.6 +/- 0.3. Aortic valve prolapse was present in 63 patients (45%), aortic insufficiency was present in 25 (18%), and 5 (3.5%) had prior bacterial endocarditis. There were no early or late deaths or major morbidity. No patient required a ventriculotomy to accomplish ventricular septal defect closure. Mean postoperative intensive care unit stay was 1.3 +/- 0.9 days, and mean hospital stay was 5.5 +/- 1.9 days. There were no instances of permanent complete atrioventricular dissociation, reoperations for bleeding, postoperative wound infections, or reoperations for residual or recurrent ventricular septal defect. These improved results justify a reevaluation of historic indications for ventricular septal defect closure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239794 TI - Which techniques of cardioplegia prevent ischemia? AB - One hundred seven patients undergoing coronary artery bypass grafting were randomized to receive warm antegrade (n = 21), warm retrograde (n = 22), cold antegrade (n = 20), cold retrograde (n = 22), or intermittent cold antegrade (n = 22) blood cardioplegia. Myocardial oxygen consumption and lactate production, adenine nucleotides, and adenine nucleotide degradation products were measured during the operation, and creatine kinase-MB release was assessed postoperatively. Warm cardioplegia resulted in greater myocardial lactate production than cold cardioplegia (p = 0.048). Retrograde cardioplegia was associated with greater lactate production than antegrade cardioplegia (p = 0.015). Adenosine triphosphate depletion was similar among groups. However, poorly diffusible metabolites of adenosine triphosphate accumulated to the greatest extent in the intermittent cold group. Levels of hypoxanthine were highest after warm retrograde cardioplegia. Operative mortality and morbidity were low and were not different among groups. In summary, none of the five techniques of cardioplegia evaluated in this study was able to completely prevent myocardial ischemia. Anaerobic lactate production was minimized with cold cardioplegia and with antegrade cardioplegic delivery. Hypothermia may have impaired regeneration of adenosine triphosphate, however, particularly in association with inadequate or intermittent cardioplegic flow. PMID- 8239795 TI - Simultaneous tracheal and esophageal pH monitoring: investigating reflux associated asthma. AB - Aspiration of gastric acid into the trachea may cause asthma in some patients who have gastroesophageal reflux. Antireflux surgery has been advocated for such patients, but lack of an objective test for acid aspiration makes patient selection difficult. We report a new technique for demonstrating acid aspiration, simultaneous tracheal and esophageal pH monitoring. Tracheal pH was measured with a 1.0-mm pH electrode introduced through the cricothyroid membrane under bronchoscopic vision. A standard esophageal pH electrode was placed in the usual position. Tracheal and esophageal pH were monitored over a 24-hour period. Peak expiratory flow rate was measured hourly while the patient was awake. We present data obtained in 3 patients with severe asthma and symptomatic gastroesophageal reflux. All 3 patients demonstrated a decrease in tracheal pH to less than 5.5, coinciding with a decrease in esophageal pH to less than 4.0. The test was repeated after antireflux operation and showed that significant decreases in esophageal pH no longer lowered tracheal pH. Asthmatic symptoms were improved, and medication was reduced in 2 of the 3 patients. PMID- 8239796 TI - Effect of sodium nitroprusside on paraplegia during cross-clamping of the thoracic aorta. AB - Sodium nitroprusside (SNP) is usually used to control excessive proximal pressure after aortic cross-clamping. To assess the effect of SNP on circulatory dynamics, somatosensory evoked potentials, and neurologic outcome, 10 adult mongrel dogs that underwent 45 minutes of cross-clamping of the thoracic aorta were randomly assigned to receive either 50 mg/kg of SNP or no treatment for excessive proximal hypertension. There was a statistically significant difference noted between the SNP-treated animals and the control animals in terms of the proximal mean arterial pressures (112 +/- 13 versus 142.2 +/- 15 mm Hg, respectively; p < 0.05) and the mean distal arterial pressures (15 +/- 3 mm Hg versus 23 +/- 1 mm Hg; p = 0.04). However, the electrical activity of the spinal cord, as indicated by the somatosensory evoked potentials, returned significantly faster in the nontreated group than in the SNP-treated group (15 +/- 9 versus 44 +/- 13 minutes; p < 0.05). Control animals exhibited a significantly better neurologic outcome and no paraplegia 24 hours postoperatively. We conclude that the use of SNP to treat excessive proximal hypertension may be detrimental to the spinal cord during cross-clamping of the thoracic aorta, resulting in a decline in the ischemic tolerance. PMID- 8239797 TI - Video Assisted Thoracic Surgery Study Group data. AB - The Video Assisted Thoracic Surgery Study Group was formed to collect data on thoracoscopic procedures in an effort to define the role for this new technique. With more than 40 participating institutions, 1,820 cases have been collected through December 1992. Lung nodules and pleural effusions represent the most frequent indications, and wedge resection and operation in the pleural space were the most common procedures performed. Four hundred thirty-nine procedures (24%) were converted to a thoracotomy because of the need for more extensive resection (219), inability to find the pathology (65), too large a lesion or difficult location (62), adhesions (58), equipment failure (25), or bleeding (10). Prolonged air leak (> 5 days) was the most frequent complication. Patients undergoing video-assisted thoracic surgery and wedge resection had a mean hospital stay of 5.1 days; video-assisted thoracic surgical lobectomy was performed in 38 patients, who had a mean stay of 6.3 days. Air leaks were more prevalent in patients with poor pulmonary function (forced expiratory volume in 1 second < 1 L) and the elderly (> or = 75 years) after video-assisted thoracic surgical wedge resection. Video-assisted thoracic surgery appears safe and may be advantageous for some procedures. Further studies will be required to define its precise role in thoracic surgery. PMID- 8239798 TI - Chronic stimulation enhances extramyocardial collateral blood flow after a cardiomyoplasty. AB - We have previously demonstrated that collateral blood flow can be established between skeletal muscle and myocardium in animals that have undergone a latissimus dorsi cardiomyoplasty. We have also shown that 5 minutes after the thoracodorsal nerve is electrically stimulated at 1.25 Hz, there is a sixfold increase in the collateral blood flow between the latissimus dorsi and the heart. In this experiment, we hypothesized that chronic stimulation of a latissimus dorsi cardiomyoplasty would result in a sustained increase in the latissimus derived collateral blood flow. In 24 adult male goats, an ameroid constrictor was placed around a branch of the circumflex coronary artery, and a latissimus dorsi cardiomyoplasty was performed. After a rest period of about 1 week, the latissimus dorsi cardiomyoplasties were stimulated continuously at a 2-Hz frequency for 6 weeks. Collateral blood flow between the muscle and the heart was then measured with colored microspheres. Sixteen animals survived to the final experiment, and collaterals developed in 10. In these 10 animals, the latissimus collaterals continuously delivered 0.17 +/- 0.03 mL.g-1 x min-1 (mean +/- the standard error) of blood to ischemic myocardium. This flow represents 24.0% +/- 3.9% of the flow measured to normal myocardium. These results demonstrate that in an animal model of coronary artery disease, chronic electrical stimulation of a latissimus dorsi cardiomyoplasty maintains an elevated level of latissimus derived collateral blood flow to the myocardium. PMID- 8239799 TI - Surgical relief of acute airway obstruction due to primary tuberculosis. AB - Primary pulmonary tuberculosis in children remains a leading cause of mortality and morbidity in developing countries. Thirteen children requiring urgent thoracotomy for relief of acute respiratory distress resulting from critical major airway narrowing caused by enlarged tuberculous mediastinal lymph nodes were admitted to two hospitals over a 4-year period. Ages ranged from 2 months to 10 years. The condition of each patient had deteriorated despite appropriate antituberculosis therapy and an oral corticosteroid. At operation, the enlarged tuberculous subcarinal or paratracheal lymph nodes or both were decompressed. Surgical complications included a bronchial tear and a pulmonary artery laceration. Additional procedures included a right upper lobectomy, two pneumonectomies, plication of a hemidiaphragm, and mobilization of two muscle flaps. Postoperatively all children showed dramatic improvement. The trachea to main bronchi diameter ratio improved by 49.1% on the left and 44.9% on the right in the immediate postoperative period. In children with respiratory distress produced by compression of the main bronchi between enlarged subcarinal and paratracheal lymph nodes, surgical decompression of the lymph nodes is indicated if there is no marked initial response to appropriate medical therapy. At operation, lymph nodes should be decompressed only by incision and curettage. Attempts at lymph node excision are associated with increased complications. PMID- 8239800 TI - A fifteen-year wound surveillance study after coronary artery bypass. AB - Wound infections after coronary artery bypass operations have been continuously monitored at the Minneapolis Veterans Affairs Hospital for 15 years. All patients were followed up for 30 days. From 1977 to 1991, 2,402 coronary artery bypass operations were performed, and wound infections developed in 125 (5%) patients. There were 71 (3%) chest infections of which 33 (1.4%) were major and 38 (1.6%) superficial. Greater than 94% of these grew only a single organism, of which 74% were Staphylococcus species. There were 63 (2.6%) leg wound infections. More than 50% of these grew multiple organisms, of which 68% were enteric in origin. Nine (0.4%) patients had simultaneous chest and leg infections. Wound infections were diagnosed an average of 15.3 +/- 6.7 (range, 4 to 30) days postoperatively, with 50% occurring after discharge from the hospital. Of 14 variables evaluated by multivariate logistic regression analysis, only steroids (p = 0.005) and diabetes (p = 0.003) were identified as independent risk factors for wound infections. Patients taking steroids or with diabetes tended to have chest infections, whereas obese patients tended to have more leg infections (p = 0.08). During an interval in the surveillance program, a trend toward increasing infections was identified and successfully reversed. PMID- 8239801 TI - Inhibition of neutrophil adhesion reduces myocardial infarct size. AB - Neutrophil accumulation and activation within the myocardium during ischemia and reperfusion has been shown to play a prominent role in the development of myocardial stunning and infarction. To determine if a simple inhibitor of neutrophil adhesion could reduce myocardial infarct size, we administered NPC 15669 (a new antiinflammatory agent that inhibits neutrophil adhesion) to 12 pigs (6 controls, 6 NPC-treated) in a porcine model of ischemia and reperfusion injury. Each animal received a continuous infusion of either NPC (10 mg/kg intravenous bolus followed by 6 mg.kg-1 x h-1 intravenous infusion) or an equal volume of normal saline solution during 1 hour of left anterior descending artery occlusion and 2 hours of reperfusion. There were no significant differences in the pre-ischemia, mid-ischemia, or postischemia rate-pressure product between control and experimental groups. The regions at risk were similar in both groups. However, the mean myocardial infarct size was reduced by 51% with administration of NPC 15669 (30.7% +/- 6.8%) compared with controls (62.3% +/- 5.4%; p < 0.01). These data indicate that NPC 15669, an inhibitor of neutrophil adhesion, substantially reduces myocardial infarct size after transient left anterior descending artery occlusion and that adhesion of the white cell to vascular endothelium may be an important element of the pathogenesis of myocardial infarction. PMID- 8239802 TI - Noninvasive detection of acute rejection in a new experimental model of heart transplantation. AB - We have shown that positron emission scintigraphy detects changes in the uptake of 18-F 2-deoxyglucose and 13-N ammonia by the acutely rejecting myocardium in a nonworking model of heterotopic heart transplantation in the rat. We developed a new working model of heterotopic heart transplantation to determine the possible relevance of these changes to clinical transplantation. Moderate aortic valvular regurgitation was produced allowing the heterotopic left ventricle to fill and eject. Rejecting allografts and nonrejecting isografts (controls) were studied 4 days after transplantation. Histologically, isografts were normal and all allografts showed mild acute rejection. Decay-corrected uptakes of 18-F 2 deoxyglucose and 13-N ammonia reflect glucose metabolism and blood flow, respectively. Values are presented as percent of injected dose per gram of tissue. Uptake of 18-F 2-deoxyglucose was higher in rejecting allografts compared with nonrejecting isografts (3.0 +/- 1.8 versus 1.1 +/- 0.4; p = 0.024). Ammonia uptake was elevated in allografts compared with isografts (2.2 +/- 0.5 versus 1.3 +/- 0.5; p = 0.023). Uptakes of 18-F 2-deoxyglucose and 13-N ammonia are higher in mildly rejecting allografts, implying increased glucose utilization and blood flow during acute rejection. These data support our earlier findings of changes in myocardial metabolism in the absence of diminishing blood flow in acutely rejecting hearts. This model may lead to a better understanding of the physiology and metabolism of acute rejection. PMID- 8239803 TI - Intercostal artery: histomorphometric study to assess its suitability as a coronary bypass graft. AB - To assess potential suitability of the intercostal artery as a conduit in myocardial revascularization a histomorphometric study of the fifth intercostal artery (n = 11) and intercostal arteries other than the fifth (n = 9) was conducted. All arteries were harvested at autopsy in 11 individuals (mean age, 75 years). The length of the intercostal arteries varied from 22 to 35 cm (mean, 29.4 cm). Three combinations of histologic patterns were seen along the course of the intercostal artery: a proximal elastic segment followed by subsequent elastomuscular and muscular segments (n = 3), a proximal elastomuscular segment with the remainder of the artery being muscular (n = 6), and a completely muscular pattern (n = 2). The mean luminal diameter of the fifth intercostal arteries varied from 1.4 +/- 0.3 mm at the origin to 0.9 +/- 0.2 mm at 30 cm (30% to 40% decrease in diameter due to the flaccid state and rigor mortis). The mean intimal thickness at these locations was 54 +/- 38 microns and 25 +/- 16 microns and the mean thickness of the media was 205 +/- 38 microns and 70 +/- 45 microns. The histologic findings, mean luminal diameter, and mean diameter of the intima and the media were similar in the intercostal arteries other than the fifth. Our study demonstrates that the intercostal artery has a relatively thin intima and media, the latter being elastic or elastomuscular in the proximal segment in the majority of the investigated arteries, which are favorable properties with regard to its potential suitability as a coronary bypass conduit. PMID- 8239804 TI - Clinical experience with the right gastroepiploic artery in coronary artery bypass grafting. AB - The excellent long-term results with the internal mammary artery for coronary artery bypass grafting have prompted the search for other conduits with similar characteristics. From December 1989 to December 1991, the right gastroepiploic artery (RGEA) has been used as an in situ graft to the posterior descending coronary artery in 51 patients at the Montreal Heart Institute. The patients' age averaged 50 +/- 11 years. Three-vessel coronary artery disease was present in 41 patients and two-vessel disease in the remaining 10 patients. In all but 1 patient, bilateral internal mammary artery grafting was performed in addition to RGEA grafting. The number of grafts per patient averaged 3.2 +/- 0.8. There was no operative mortality. Morbidity was minimal with only myocardial infarction and a pleural effusion in 1 patient. In 1 patient, a splenectomy had to be performed because of iatrogenic tear during dissection of the RGEA. The average hospital stay was 8.2 +/- 2.6 days. Enteral nutrition was resumed on average 2 days after operation. Angiographic evaluation of RGEA grafts was performed before discharge or within the first month after surgery in 31 patients. In 28 patients (28/31, 90%) the RGEA graft was patent, two grafts were occluded, and in the remaining patient, the graft could not be visualized due to technical difficulties during angiography. A second angiographic evaluation was performed in 5 patients, 1 year after operation. Four RGEAs were patent and 1 was occluded. Clinical follow-up averaged 4 months (range 1 to 15 months).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239805 TI - Different vascular reactivity of human internal mammary and inferior epigastric arteries in vitro. AB - Vascular responses to endogenous agonists may determine patency rates of bypass graft conduits. The effect of constrictors (noradrenaline, phenylephrine, serotonin, histamine, angiotensin II) and dilators (acetylcholine, substance P, bradykinin, nitroglycerin) were compared in human internal mammary and inferior epigastric arteries in vitro. The latter vessel type has been recently advocated as an additional conduit for coronary artery bypass grafting. Whereas the alpha adrenoceptor- (noradrenaline, phenylephrine) and serotonin receptor-mediated contractions were similar in both vessels, histamine-induced contractions were greatly enhanced in internal mammary arteries (maximal responses in percent of 80 mmol/L KCl, 131% +/- 15% versus 59% +/- 8%). Maximal contractions in response to angiotensin II were greater in inferior epigastric arteries (50% +/- 6% versus 25% +/- 5%). The endothelium-independent relaxations in response to nitroglycerin were identical in both vessels. In contrast, the endothelium-dependent relaxations in response to acetylcholine, substance P, and bradykinin were significantly greater in the inferior epigastric than in the internal mammary arteries (maximal relaxations expressed as percent of prostaglandin F2 alpha induced precontraction: acetylcholine, 94% +/- 5% versus 77% +/- 5%; substance P, 85% +/- 4% versus 24% +/- 5%; bradykinin, 77% +/- 5% versus 26% +/- 3%). It is concluded that the inferior epigastric artery has a high endothelial capacity to release endothelium-derived relaxing factor. It appears that the inferior epigastric artery possesses credentials to be successfully used for coronary artery bypass grafting. PMID- 8239806 TI - Vasoconstrictor profile of the inferior epigastric artery. AB - The inferior epigastric artery is a putative arterial bypass graft. The receptor mechanisms that control vascular tone are thought to play a role in the performance of bypass conduits. We have compared the vascular reactivity of the inferior epigastric artery with that of the internal mammary artery. Segments from a total of 15 inferior epigastric and 12 internal mammary arteries were examined for their response to increasing concentrations of noradrenaline, 5 hydroxytryptamine, dopamine, histamine, endothelin-1, or the thromboxane analogue U46619. The responsiveness of the smooth muscle was significantly greater in the inferior epigastric artery (p < 0.05) as judged by contractions elicited by 90 mmol/L potassium chloride. However, although the response of the inferior epigastric artery tended to be greater, this significant enhancement of smooth muscle function was not paralleled by the maximal responses of noradrenaline, 5 hydroxytryptamine, dopamine, histamine, or endothelin-1. However, the tension generated in response to U46619 did differ significantly, with maximal responses in the inferior epigastric and internal mammary arteries of 59.2 +/- 8.3 mN and 35.0 +/- 3.6 mN, respectively. When receptor function was compared by expressing the response as a percentage of that of 90 mmol/L potassium chloride, it was revealed that noradrenaline was capable of inducing significantly greater relative contractions in the internal mammary artery (114.8% +/- 20.5%) as compared with the inferior epigastric artery (49.9% +/- 19.1%); the potency of this constrictor was sixfold greater in the internal mammary artery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239807 TI - Mediastinal paraganglioma: a surgical experience. AB - Between October 1960 and January 1991, 5 male and 9 female patients (mean age, 44 +/- 19 years) were seen at the Mayo Clinic because of mediastinal paraganglioma. Nine patients presented with hypertension. In these and an additional patient, the tumors were biochemically active. In 13 patients, the mediastinal mass was localized preoperatively. All patients with functioning tumors received alpha adrenergic and beta-adrenergic blockade preoperatively. Total resection was performed in 11 patients and partial resection in 2. In 1 patient, the tumor was found at postmortem examination. Five lesions were grossly invasive and nine were noninvasive. Blood pressure returned to normal in 6 of the 8 hypertensive patients whose tumors were totally removed. In 4, catecholamine excretion normalized as well. Tumor multicentricity occurred in 6 patients. In 7 patients, the disease took a malignant course (metastases in 5 and progression of unresectable lesions in 2). The probability of 10-year survival after resection was 50% in the group with malignant tumors, and there were no deaths in the group with benign lesions. Analysis of the DNA cellular content showed no correlation between the ploidy pattern and the behavior of the disease. PMID- 8239808 TI - Predictors of early saphenous vein aortocoronary bypass graft occlusion. The GESIC Study Group. AB - To identify factors determining early saphenous vein aortocoronary bypass occlusion, we analyzed the data base of the GESIC study, a trial comparing antiplatelet drug regimens that included 927 patients with 1,854 saphenous vein grafts. The univariate analysis showed female sex (p < 0.0097), obesity (p < 0.001), rest angina (p < 0.0026), history of congestive heart failure (p < 0.037), the revascularized artery (p < 0.0001), the quality of distal bed (p < 0.00001), the diameter of the grafted vessel (p < 0.00001), the lack of antiaggregant treatment (p < 0.017), and a nonsequential technique (p < 0.0002) as predictors of early (28 days) graft occlusion. In the multivariate analysis the last five variables were independent predictors. Using the two preoperative variables, it was possible to identify groups at different risk; the occlusion rate ranged between 8.79% (left anterior descending coronary artery and good distal vessel) and 27.58% (right coronary artery or left circumflex coronary artery and poor distal vessel). The combination of three variables (grafted vessel, artery diameter, and antiaggregant treatment) also allowed identification of different risk groups; the occlusion rate ranged between 3.5% and 63.1%. Thus, it is possible to anticipate the risk of graft occlusion in patients undergoing coronary artery bypass grafting, which may help in the selection of both patients and antithrombotic treatment. PMID- 8239809 TI - Simultaneous total aortic replacement from valve to bifurcation: experience with 21 cases. AB - From June 1985 to December 1991, 21 patients (12 men and 9 women; mean age, 60 years) underwent total simultaneous aortic replacement that extended from the valve to the bifurcation. The causes of the diseased aorta were: medial degeneration with total aortic dilatation or multiple aneurysms (n = 7) and either acute (n = 4) or chronic (n = 10) dissection. Clinical evaluation and investigation in all patients consisted of computed tomography and magnetic resonance imaging as well as angiography. Only patients with combined thoracic and abdominal emergencies were selected, and these comprised worsening of cardiac conditions resulting from aortic regurgitation, and rapid dilatation of the ascending aorta and arch with impending rupture in conjunction with ischemia of the abdominal viscera, kidney, or either leg. The surgical technique consisted of inducing deep hypothermia by means of femoral vein-femoral artery cardiopulmonary bypass. During the cooling time, the aortic root was replaced under cardioplegia. Once lowering of the body temperature attained electroencephalographic silence, circulation was stopped and the aorta was replaced from the arch to the bifurcation. Circulation and rewarming were resumed only after the operation was completed. In our most recent patient, the operating time was reduced by opening the thoracic and the abdominal incisions during cooling; the cardioplegic solution as not injected but, instead, the myocardium was cooled down along with the whole body. In these patients, the hypothermy at electroencephalographic silence ranged from 14 degrees to 19 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239810 TI - Degradation of a supporting prosthesis can optimize arterialization of autologous veins. AB - In a previous study, we implanted autologous vein grafts in the carotid artery of rabbits supported by a compliant, biodegradable prosthesis to prevent vein wall damage due to the higher arterial pressure. We showed that such a supporting prosthesis indeed reduces damage to these vein grafts and allows for more regular and gradual arterialization than that afforded by unsupported vein grafts. To evaluate the influence of the rate of biodegradation of such a supporting prosthesis on the process of arterialization of autologous vein grafts, we implanted vein grafts supported with prostheses, which degrade within 3 weeks (group I), 6 weeks (group II), or 3 months (group III), into the carotid artery of rabbits, and then evaluated them up to 6 weeks after implantation. At 6 weeks, the group I vein grafts showed a thinner vein wall than did the adjacent artery during dilatation. In group II, the vein wall thickness and luminal diameter had completely adjusted to that of the adjacent carotid artery. The group III vein grafts showed a significantly thinner vein wall in the absence of dilatation. All supported vein grafts showed regular longitudinally oriented and, in some areas, circularly oriented cell layers, together with thin elastic laminae, which were most pronounced in group II. We conclude that a supporting, compliant prosthesis can stimulate, regulate, and optimize the arterialization of autologous vein grafts in rabbits. If the rate of degradation is carefully chosen, the radius and wall thickness of the vein graft can completely adjust to that of the adjacent artery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239811 TI - Respiratory dysfunction after uncomplicated cardiopulmonary bypass. AB - Respiratory dysfunction is a well-recognized complication of cardiac operations. To quantify its current incidence and severity after uncomplicated cardiopulmonary bypass, serial measurements of arterial oxygen tension (PaO2), alveolar-arterial oxygen gradient (AaO2), and percentage pulmonary shunt fraction (%PSF) measured by a noninvasive technique were made in 129 patients (age, 59 +/- 8 years (mean +/- standard deviation) with good left ventricular function (left ventricular end-diastolic pressure < 15 mm Hg) undergoing isolated coronary artery operations (group 1) and 30 patients undergoing general surgical procedures (group 2). Measurements were made before operation and on the first, second, and sixth postoperative days. Seven patients in group 1 who required prolonged ventilation were excluded from further study. In group 1, between the preoperative and second postoperative days, there was a marked fall in PaO2 [89 +/- 11 versus 57 +/- 9 mm Hg; p < 0.001] and a marked increase in the AaO2 gradient [18 +/- 10 versus 50 +/- 11 mm Hg; p < 0.001)] and %PSF [3 +/- 1% versus 19 +/- 6%; p < 0.001)] with only modest improvement by the sixth postoperative day [PaO2, 67 +/- 11 mm Hg; AaO2, 45 +/- 11 mm Hg; %PSF, 15 +/- 4]. There were similar but less severe changes in PaO2 and AaO2 gradients in group 2 patients, with a return to baseline values by day 6.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239812 TI - Clinical and hemodynamic performance of the 19-mm Carpentier-Edwards porcine bioprosthesis. AB - Because of concerns about the hemodynamic performance of 19-mm porcine valves, we retrospectively reviewed the clinical results and echocardiographic studies of 52 consecutive patients who received a 19-mm Carpentier-Edwards porcine bioprosthesis (model 2625) for aortic valve replacement from 1986 through 1991. Nearly 87% of the patients were women, the mean age was 69 years, and the mean body surface area was 1.63 +/- 0.27 m2. Seventy-three percent of the patients had pure aortic stenosis, 96% were in New York Heart Association classes III and IV, and 56% underwent urgent or emergent operation. Overall hospital mortality was 7.7% with a late mortality of 8.3% at a mean follow-up of 25 +/- 18 months. No patient experienced a valve-related complication, and 95% of surviving patients were in New York Heart Association classes I and II. Two-dimensional and Doppler echocardiography performed during the first postoperative week revealed a maximal instantaneous gradient of 44.7 +/- 13.0 mm Hg. In 43 patients for whom additional data were available, the mean gradient was 26.4 +/- 8.2 mm Hg with an effective orifice area of 0.85 +/- 0.18 cm2. This study defines the normal range of postoperative gradients across the 19-mm Carpentier-Edwards porcine valve and demonstrates that patients receiving this valve can achieve significant clinical improvement despite the presence of high transvalvular gradients measured by echocardiography. PMID- 8239813 TI - Limitation of vasodilation associated with warm heart operation by a "mini cardioplegia" delivery technique. AB - Peripheral vasodilation is commonly seen during and after warm heart operations and can become of clinical concern when it requires vasopressors because some of these drugs adversely affect coronary artery bypass graft flows. As hemodilution lowers systemic vascular resistance, we assessed whether peripheral vasodilation could be limited by a drastic reduction of the volume of infused cardioplegia. Fifty patients underwent isolated coronary artery bypass grafting procedures using normothermic (35 degrees to 37 degrees C) bypass and normothermic continuous retrograde blood cardioplegia. They were divided into two equal groups: in group 1, blood was diluted 4:1 with hyperkalemic crystalloid cardioplegia, whereas in group 2, the cardioplegic "solution" was limited to the sole arresting agents that were concentrated in a small volume (16 mEq potassium chloride and 3 mEq magnesium chloride in a 20-mL ampoule). This "mini cardioplegia" was continuously added to arterial blood so as to keep the heart arrested. The average volume of cardioplegia per patient was 1,000 mL in group 1 and 58 mL in group 2 (p < 0.0001). The mini-cardioplegia technique resulted in a reduced incidence of perioperative systemic vasodilation: group 2 patients required significantly less vasopressors (p < 0.05) and less volume loading, as reflected by significantly lower right atrial and pulmonary capillary wedge pressures (p < 0.05 and p < 0.03 at 12 hours postoperatively, respectively), compared with group 1 patients who received traditional high-volume cardioplegia. There were no differences between the two groups with respect to myocardial recovery, as assessed by standard clinical and hemodynamic end points.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239814 TI - Postischemic deterioration of sarcoplasmic reticulum: warm versus cold blood cardioplegia. AB - Impaired cardiac sarcoplasmic reticulum (SR) function, as evidenced by reduced SR Ca2+ uptake rate and decreased SR Ca(2+)-adenosine triphosphatase activity, has been found in postischemic "stunned" myocardium and in hearts subjected to hypothermic arrest. In this study, we compared the effects of retrograde continuous coronary sinus warm blood cardioplegia (WBC) and retrograde intermittent cold blood cardioplegia (CBC) on cardiac SR function and postischemic ventricular functional recovery in pig hearts. Twelve in situ isolated pig hearts supported by cardiopulmonary bypass were subjected to 120 minutes of cardioplegic arrest with either WBC (37 degrees C) or CBC (6 degrees to 10 degrees C), followed by 60 minutes of 37 degrees C reperfusion. Left ventricular global contractile function and coronary blood flow were measured before arrest and during reperfusion. Cardiac SR was isolated from left ventricular biopsy specimens, and 45Ca2+ uptake by SR and SR Ca(2+)-adenosine triphosphatase activity were determined. The recovery of left ventricular global contractile function as indicated by the maximum of the first derivative of left ventricular pressure was significantly improved in the WBC group compared with that of the CBC group (70% versus 46%; p < 0.05). The SR Ca(2+)-adenosine triphosphatase activity was better preserved after 60 minutes reperfusion in WBC compared with CBC (0.31 +/- 0.02 versus 0.20 +/- 0.03 microM Pi/min/mg protein, p < 0.05), and the recovery of SR Ca2+ uptake was significantly improved by WBC compared with CBC (1.15 +/- 0.12 versus 0.83 +/- 0.04 microM Ca2+/min/mg protein; p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239815 TI - Surgical resection of a second primary lung carcinoma in a survivor of small cell carcinoma. AB - Resection of a second primary lung carcinoma in a long-term survivor of small cell lung carcinoma is reported. The patient had adenocarcinoma as the second primary tumor and underwent lobectomy with mediastinal node dissection. He was alive 99 months after the initial diagnosis of small cell carcinoma and 54 months after curative pulmonary resection for his second primary carcinoma. PMID- 8239816 TI - Infective aortic endocarditis after percutaneous balloon aortic valvuloplasty. AB - Infective aortic endocarditis developed in an elderly patient after a percutaneous balloon aortic valvuloplasty. The transesophageal echocardiogram demonstrated a perivalvular abscess. The patient underwent surgical replacement of the infected valve, but later succumbed to renal failure. The development of infective aortic endocarditis should be recognized as a potentially fatal complication of percutaneous balloon aortic valvuloplasty. The important measures in preventing bacteremia during percutaneous balloon aortic valvuloplasty and the appropriate role of operation are discussed. PMID- 8239817 TI - Reconstructing an anomalous left coronary artery origin using the internal iliac artery. AB - We describe reconstruction of an anomalous origin of the left coronary artery from the pulmonary artery. The right internal iliac artery was used to create the trunk. The procedure was carried out using extracorporeal circulation during normothermia with a beating heart. PMID- 8239818 TI - Leiomyoma of the bronchus treated by endoscopic resection. AB - Leiomyoma of the bronchus is a rare benign tumor. The type of surgical resection depends on the location of the tumor and secondary lung destruction, although most surgical treatment has required thoracotomy. Two Korean women who had pedunculated masses in the left main bronchus and in the right lower lobe bronchus were successfully treated by endoscopic resection. Their postoperative courses were uneventful. They are well 19 months and 10 months, respectively, after resection. PMID- 8239819 TI - Transposition of the great arteries: correction with conversion from single to dual coronary artery system. AB - We describe the case of a newborn child with D-transposition of the great arteries who was found to have an uncommon single-origin coronary artery system. Surgical correction by arterial switch was performed but resulted in tension of the left coronary branch, which prevented discontinuation of cardiopulmonary bypass. The problem was eventually resolved by conversion into a dual coronary artery system through direct implantation of the left coronary branch into the aorta. The patient remains well, free from ischemia and with a patent left coronary artery, 2 years after operation. PMID- 8239820 TI - Anomalous origin of the left coronary artery from the pulmonary artery. AB - A 20-year-old woman underwent successful operative correction of anomalous origin of the left coronary artery from the pulmonary artery by left internal mammary artery bypass graft. The clinical, hemodynamic, and angiographic features of this unusual case are presented. Different therapeutic options are discussed. PMID- 8239821 TI - Stentless xenograft valve in a truncus arteriosus communis. AB - We report the use of a recently developed stentless porcine valve in a 2-year-old child with truncus arteriosus communis (type I). The child had received no previous surgical palliation. Despite a markedly increased pulmonary arteriolar resistance the right ventricle was connected to the pulmonary artery with a homograft. The postoperative course was complicated at first by right ventricular failure due to pulmonary hypertension and secondarily by left ventricular failure caused by rapidly increasing truncal valve incompetence. Because we did not have a suitable homograft at hand we replaced the truncal valve with a stentless porcine xenograft valve. PMID- 8239822 TI - Surgical repair of atrial septal rupture due to blunt trauma. AB - We report a case of atrial septal rupture and surgical cure after blunt chest trauma. Review of the literature indicates that this rare lesion results from severe forces applied to the chest and is often associated with other serious and life-threatening injuries. The defect may not be recognized for several months or even years in patients who survive concomitant initial trauma. When operative repair is undertaken, a favorable outcome can be anticipated. PMID- 8239823 TI - Heart transplantation for tumor. AB - Unresectable cardiac tumors, although unusual, are often rapidly fatal. A 31-year old woman presented with a large tumor arising from the left ventricle and causing symptoms of a constrictive cardiomyopathy. After evaluation with echocardiography, angiography, and computed tomography, an exploration was carried out to confirm the extent of disease. Orthotopic heart transplantation was subsequently performed when a donor organ became available. She is now alive and disease-free 12 months after transplantation. PMID- 8239824 TI - Clinical cardiopulmonary bypass without systemic anticoagulation. AB - Heparin-coated perfusion systems have been used to perform cardiopulmonary bypass with decreased systemic heparin dosages. We report the case of a 60-year-old man with end-stage liver disease and severe aortic stenosis who underwent an aortic valve replacement without systemic anticoagulation, employing a heparin-coated perfusion system. In patients with a contraindication to systemic anticoagulation, cardiopulmonary bypass with a heparin-coated system without systemic anticoagulation may be a viable alternative. PMID- 8239825 TI - Pedunculated pulmonary leiomyoma with large cyst formation. AB - A case of single pulmonary leiomyoma with giant cyst formation is presented. Metastasizing uterine fibroleiomyoma and fibroleiomyomatous hamartoma both give rise to multiple pulmonary nodules on chest films. Leiomyoma of the lung presenting as a single pedunculated lesion with cyst formation is exceptional. This report documents the existence of other rare cystic lesions that may mimic the more common cystic air space and bullous disease. PMID- 8239826 TI - Atrial myxomas in young children: an alternative surgical approach. AB - A case report and a brief review of the literature on atrial myxomas in young children (5 years and younger) are presented. Right atrial myxomas are predominant in this age group in contrast to older children and adults. The echocardiogram alone can provide clear anatomic information for the operation. For large left atrial myxomas, wide exposure through a transverse biatrial incision is recommended to avoid tumor manipulation and facilitate en bloc resection. PMID- 8239827 TI - New shapes and sizes: a T-tube coronary conduit. AB - A novel type of coronary artery graft named a T-tube conduit is described. This reversed saphenous graft, coming in three different variations, is useful at long arteriotomies, at revision redo anastomoses, and when size mismatch exists between vein and artery. PMID- 8239828 TI - The first successful surgical treatment of mitral stenosis: the 70th anniversary of Elliot Cutler's mitral commissurotomy. AB - In 1923 Elliot Carr Cutler, in conjunction with his cardiology colleague, Samuel Levine, performed a closed transventricular mitral commissurotomy with a tenotomy knife on a 12-year-old patient dying of rheumatic mitral stenosis at the Peter Bent Brigham Hospital. This operation was carried out after several years of experimentation regarding resuscitation of the heart, appropriate incisions, and the pathophysiology of mitral stenosis. The interest in mitral stenosis was rampant at the time because of the huge number of patients suffering from this public health problem. The patient survived and went on to die of pneumonia 4 years postoperatively. Subsequent to this, Cutler performed seven more operations using his new cardiovalvulotome, which was to create controlled mitral regurgitation. Unfortunately, this concept did not promote long-term success and a moratorium for these operations was called in 1929. Nevertheless, this pioneering effort in 1923 was the first successful operation to treat valvular heart disease by a surgical technique. PMID- 8239829 TI - Barrett's disease: pathophysiology of metaplasia and adenocarcinoma. AB - Peptic ulceration arising in the lower esophagus lined by columnar epithelium was described in detail by Tileston in 1906. Although this concept was challenged in 1950 by Barrett, experimental and clinical evidence has now conclusively demonstrated that Barrett's metaplasia is an acquired condition and is a consequence of chronic reflux of gastric or duodenal contents or both. Current concepts suggest that unknown trophic factors present in these secretions stimulate proliferation of multipotential reserve cells located in the esophageal submucosal glands resulting in columnar metaplasia of the normal squamous epithelium with subsequent potential for malignant degeneration. Today, numerous patients are affected by reflux esophagitis, a lesser number by Barrett's metaplasia, and a smaller but ever-enlarging group by adenocarcinoma. Although high-grade dysplasia is considered a precursor to invasive adenocarcinoma, detection of this abnormal mucosa remains controversial and currently requires esophagoscopy with biopsy. Epithelial markers, such as increased activity of mucosal ornithine decarboxylase, sulfomucin production, nuclear DNA aneuploidy, and recently molecular analysis, have also been proposed to identify those paitents at increased risk for malignant degeneration. As more is learned about the pathogenesis of Barrett's disease, perhaps these cancers can ultimately be prevented. PMID- 8239830 TI - Chest trauma. PMID- 8239831 TI - Report of the Ad Hoc Committee on Physician-Specific Mortality Rates for Cardiac Surgery. PMID- 8239833 TI - Presidential address. PMID- 8239832 TI - Practice guidelines in cardiothoracic surgery. American Association for Thoracic Surgery, Society of Thoracic Surgeons, Southern Thoracic Surgical Association, Western Thoracic Surgical Association. PMID- 8239834 TI - Cardioplegia delivery during continuous normothermic cardiopulmonary bypass. PMID- 8239835 TI - Extended vertical transatrial septal approach for the removal of left atrial myxoma. PMID- 8239836 TI - Our experience with using the Pall LG6 leukocyte depleting filter during cardiopulmonary bypass. PMID- 8239837 TI - When and why CPD in continuous warm blood cardioplegia? PMID- 8239838 TI - Esophageal exclusion. PMID- 8239839 TI - Pulmonology and video surgery. PMID- 8239840 TI - Valvular disease in the elderly: influence on surgical results. PMID- 8239841 TI - Open heart operations in the elderly: changing risk parameters. AB - Greater numbers of people are living longer, causing a concurrent change in the population of patients who require open heart operations. One question that needs to be addressed is the effect of age on surgical outcome. Current statistics suggest that in 15 to 20 years there will be 12 million people in the United States more than 80 years of age. We examined risk factors for outcome in two retrospective studies of advanced-age patients requiring cardiac operations. The first group of 100 patients more than 75 years of age had valve replacement. Early mortality was acceptable at 3%. However, mortality at 2 years increased to about 25%, due largely to complications related to anticoagulation. Quality of life was dramatically improved in survivors. These data suggest that implantation of a tissue valve may be the most beneficial option in patients aged more than 70 years. In the second group, we used multivariate analysis to define the factors contributing to early mortality in 133 patients aged 80 years or older who required cardiac operations from 1988 to 1990. This analysis showed that early mortality increased significantly in patients in New York Heart Association class IV or in those who required postoperative blood transfusion. These data suggest the need to prevent excessive bleeding and to assess patients on an individual basis. PMID- 8239842 TI - Risk analysis of primary versus reoperative coronary artery bypass grafting. AB - Reoperative coronary artery bypass grafting is being performed with increasing frequency, in part as a function of the increasing pool of patients who have undergone initial coronary artery bypass grafting and in part because of the natural progression of atherosclerosis. The great majority of patients require reoperation because of graft atherosclerosis or because of a combination of graft and native-vessel disease. Significant risk factors for reoperation include the lack of an internal mammary artery graft or incomplete revascularization at the time of the primary operation, age, and New York State Heart Association classification. Despite an increasing experience with reoperations, operative mortality remains high, approximately three to five times that of initial bypass operation. Similarly, reoperations are associated with increased morbidity, including increased rates of bleeding and low output states. Specific problems encountered at reoperation include graft atherosclerosis, progression of native vessel disease, and a significant increase in perioperative bleeding. Bypass ischemic times tend to be longer. The use of retrograde cardioplegia and blood conservation programs may prove to be effective solutions for these problems. If perioperative results can be improved, it would appear that the long-term outlook for these patients is reasonably good. PMID- 8239843 TI - Is intraoperative echocardiography a useful monitor in the operating room? AB - Few cardiac surgeons or anesthesiologists would argue with the view that intraoperative echocardiography, both transesophageal and epicardial, has become an important diagnostic tool. Specific diagnostic applications primarily involve valve reconstruction and replacement, as well as complex congenital heart disease repairs. Routine echocardiographic monitoring in cardiac operations is, however, controversial. Three specific modalities may benefit from this technology. Aortic root scanning before cannulation/proximal coronary anastomoses using surface echocardiography is one. Areas with significant atherosclerosis can be identified and manipulation avoided. Another is quantification of global left-ventricular function using transesophageal echocardiography. This method may have specific relevance to patients requiring mechanical circulatory assistance. Using echocardiography to monitor ischemic regional wall-motion abnormalities and using the findings to predict an unfavorable outcome is a third suggested use for perioperative monitoring in cardiac surgery. PMID- 8239844 TI - Cerebral injury and cardiac operations. AB - Cerebral complications constitute the leading source of morbidity and disability after cardiac operations. The incidence of stroke after coronary artery bypass grafting has increased in tandem with the mean age of the patient population. Although many cerebral deficits resolve with time, others remain sources of disability for otherwise functional patients and detract from an otherwise successful procedure. The clinical spectrum of cerebral complications includes both neurologic and neuropsychologic deficits. Neurologic deficits include fatal cerebral injury, stroke, impaired level of consciousness, and seizures. The incidence of these deficits is 1% to 6%. Neuropsychologic deficits refer to cognitive changes, and are quantitated with tests of memory and learning and speed of visual-motor response. The incidence of these deficits is 60% to 80% at 1 week after operation and 20% to 40% at 8 weeks after operation. Central nervous system complications after cardiac operations have been attributed in large part to the effects of cardiopulmonary bypass on the brain. Potential mechanisms include macroembolization of air or particulate matter; microembolization of gas, fat, aggregates of blood cells, platelets or fibrin, and particles of silicone or polyvinylchloride tubing; and inadequate cerebral perfusion pressure. Methods of assessment include those applied during the procedure (clinical observation, assessment of cerebral blood flow and metabolism, intraoperative electroencephalography, transcranial and carotid Doppler echography, quantitative embolic measurement, and fluorescein angiography) and those performed to measure outcome (neurologic and neuropsychologic testing, computed tomographic scans, magnetic resonance imaging, and cerebrospinal fluid studies). Much of the literature regarding cerebral injury and cardiopulmonary bypass is descriptive, relating patient risk factors to the incidence of postoperative stroke.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239845 TI - Role of cytokines in the pathogenesis of cardiopulmonary-induced multisystem organ failure. AB - The clinical presentation of patients with multiorgan failure caused by septic conditions is very similar to that seen in patients with multiorgan failure after cardiopulmonary bypass. It has been hypothesized that the same mechanisms are at work in both instances. This commonality of presentation and mechanisms is denoted by the new term systemic inflammatory response syndrome. The systemic inflammation resulting from cardiopulmonary bypass is manifested by the development of adult respiratory distress syndrome. Overall mortality for this condition is high, and the absence of a specific therapy reflects the lack of understanding of the mechanisms involved. The risk factors associated with multiorgan failure include the age of the patient, the number of failed organs, and whether these organ failures persist or resolve. The release of a variety of inflammatory mediators has been implicated in the pathogenesis of sepsis. These include the cytokines (tumor necrosis factor, interleukin-1, interleukin-6), lipid and arachidonate metabolites, platelet-activating factor, and activation of the coagulation cascade. There seems to be marked synergy between these different mediators, suggesting that a combination of small amounts of them all may be more toxic than a large release of one by itself. During cardiopulmonary bypass, increased levels of circulating endotoxin have been associated with the activation of the complement system and increased levels of tumor necrosis factor. Interleukin-6 level has been shown to be elevated during bypass. The action of the inflammatory mediators to induce injury may be related to the activation of leukocytes and endothelial cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239846 TI - Acute fevers of unknown origin. A plea for restraint. PMID- 8239847 TI - Medical technology at the end of life. What would physicians and nurses want for themselves? AB - BACKGROUND: Advance directives assume that patients are able to decide what interventions they would wish in the event of catastrophic illness. This study examines the preferences of nurses and physicians, who have extensive exposure to sick patients, for care at the end of life. METHODS: Nursing and medical staff of a community teaching hospital were asked to complete the medical directive, detailing which of 12 interventions they would wish for themselves in each of four scenarios. Two additional scenarios were added to ascertain preferences for care in the event of severe illness in a previously healthy 85-year-old subject and in a chronically ill 75-year-old subject. RESULTS: Refusal rate among the 127 nurses and 115 physicians who completed the questionnaire, averaged over the four scenarios, was 78%. Nurses and physicians refused 31% of proposed therapies in the case of acute illness in a previously healthy 85-year-old subject and 57% of interventions in the case of major illness in a 75-year-old subject with multiple debilitating chronic illnesses. Nurses reported significantly higher refusal rates than physicians for the scenarios involving possible reversible coma, the healthy 85-year-old subject, and the chronically ill 75-year-old subject. Factors predicting refusal patterns were age and being a nurse. CONCLUSION: We conclude that physicians and nurses, who have extensive exposure to hospitals and sick patients, are unlikely to wish aggressive treatment if they become terminally ill, demented, or are in a persistent vegetative state. Many would also decline aggressive care on the basis of age alone, especially in the presence of functional impairment. These findings call into question the utility of detailed advance directives and suggest a need to focus on the goals of treatment for all elderly patients. PMID- 8239848 TI - Three-decade investigation of familial pheochromocytoma. An allele of von Hippel Lindau disease? AB - BACKGROUND: Usually sporadic, pheochromocytoma can, on occasion, complicate genetic disorders, such as neurofibromatosis 1, von Hippel-Lindau disease, and multiple endocrine neoplasia 2; some families seem to have just pheochromocytoma, where it may have occurred by chance. The natural history of a large kindred believed to have an excess of pheochromocytoma 34 years ago was followed with the hypothesis that the predisposition was, in fact, present and that family education and surveillance would decrease mortality. METHODS: Prospective observation and diagnostic surveillance for pheochromocytoma were conducted on the inception cohort, defined as three branches of the kindred in 1960. Of 619 descendants of three (of 11) siblings of German origin, 333 were evaluated in person at least once in the three decades of surveillance. No pheochromocytomas were known to have occurred in the eight other branches. A total of 522 persons from the 11 branches were evaluated. RESULTS: Five of the eight initial patients with pheochromocytoma died of cardiovascular complications attributable to the tumor. In follow-up, eight additional relatives were newly diagnosed with pheochromocytomas (at an average age of 19 years), and others had additional or recurrent pheochromocytomas, meningioma, para-adrenal paraganglioma, and a functioning glomus vagale; none died. CONCLUSIONS: A continuing excess of pheochromocytoma seems present in the family. Whether the incompletely penetrant gene in this family is allelic to the von Hippel-Lindau gene on chromosome 3 or is a distinct locus remains to be resolved with molecular studies. Meanwhile, education and surveillance seem to decrease mortality from pheochromocytoma in this family. PMID- 8239849 TI - A controlled study comparing the effects of nabumetone, ibuprofen, and ibuprofen plus misoprostol on the upper gastrointestinal tract mucosa. AB - BACKGROUND: This study was developed to compare the incidence of endoscopically diagnosed ulcers in elderly patients taking nabumetone, ibuprofen, or concomitant ibuprofen/misoprostol. Further research is indicated to better establish the clinical relevance of these endoscopy findings. METHODS: We conducted a prospective, multicenter, randomized, endoscopist-blinded, 12-week study involving 171 patients with osteoarthritis aged 60 years and older. Patients were randomized to receive nabumetone, 1000 mg (n = 58); ibuprofen, 600 mg four times daily (n = 53); or ibuprofen, 600 mg four times daily, administered concomitantly with misoprostol, 200 micrograms four times daily (n = 60). Endoscopy was performed at baseline and at weeks 2, 6, and 12. Endoscopy results were scored on a scale of 1 to 9. Significant ulcers were defined as breaks in the mucosa greater than 5 mm with appreciable depth. RESULTS: Of the 171 randomized patients, 148 completed the study. There was no significant difference in the incidence of significant ulcers between the nabumetone group and the ibuprofen/misoprostol group (one vs zero). There were significantly fewer significant ulcers in the nabumetone and ibuprofen/misoprostol groups than in the ibuprofen monotherapy group (one and zero vs eight; P < .01). There also was a significant difference in the time to ulcer development, with a greater risk of developing an ulcer sooner with ibuprofen treatment (P < .01) than either nabumetone or ibuprofen/misoprostol treatment. The severity of osteoarthritis, based on physicians' assessments, improved in 64% of patients in the nabumetone group, 55% of those in the ibuprofen group, and 63% of those in the ibuprofen/misoprostol group. CONCLUSIONS: Nabumetone is equivalent in ulcerogenicity to concomitant ibuprofen/misoprostol and is significantly less ulcerogenic than ibuprofen alone. PMID- 8239850 TI - Transfusion practice in medical patients. AB - OBJECTIVE: Blood transfusion raises serious issues of safety and economics. We therefore examined blood usage and its characteristics in medical inpatients, given the relative scarcity of existing data. DESIGN: During a 1-year period, transfusion episodes on two medical services were reviewed by five specialists for justifiability on the basis of generally agreed on guidelines. SETTING: The study was conducted at two institutions, a municipal teaching hospital where house staff deliver care and a community hospital where patients are under the direct care of private physicians. PATIENTS: Four hundred thirty-eight randomly selected transfusion episodes on the medical services of the two institutions were reviewed. MAIN OUTCOME MEASURES: The prevalence of unjustifiable transfusions based only on the information available to the managing physician at the time of transfusion. RESULTS: Eighteen percent of the 438 randomly selected transfusion episodes were viewed as not justifiable by at least four of five reviewers; another 17% were classified as equivocal because two or three reviewers judged them to be not justifiable. The most striking observation was the greater prevalence of nonjustifiable transfusion episodes at the community hospital (26% vs 16% at the teaching institution; P = .0121). Other observations included a tendency for physicians to prescribe transfusions by the numbers (at least 11% of nonjustifiable transfusions) and to overtransfuse. The routineness with which transfusion was viewed by managing physicians was also identifiable by the absence of written transfusion notes in 39% of all episodes reviewed, which incidentally raises questions about the adequacy of the medical chart's documentary functions today. CONCLUSIONS: The rate of nonjustifiable or equivocal transfusion on medical services may be as high as 35%. Reliance on numbers rather than clinical status seems to be a major problem. Education is obviously a critical issue and should also target private practitioners, who seemed to perform less well than physicians in training. Transfusion guidelines that use specific hematocrit values also need to be reexamined. PMID- 8239851 TI - Internal medicine training in ambulatory gynecology. A house of staff survey. AB - OBJECTIVES: Practicing internists commonly provide gynecologic care, yet internists receive little formal training in ambulatory gynecology. To evaluate current residency training, we surveyed house staff perceptions of their training in gynecology. METHODS: Data were collected from house staff at five Portland, Ore, internal medicine training programs by questionnaire using a Likert scale about training in 24 gynecologic and five nongynecologic internal medicine problems. Responses were compared by level of training, institution, and gender. RESULTS: Responses were received from 195 (89%) of 221 residents. House staff reported less than adequate training in gynecology, particularly in the areas of family planning and medical complications of pregnancy, while reporting much superior training in the traditional medicine problems. CONCLUSIONS: House staff in internal medicine report little training in the diagnosis and treatment of gynecologic problems. Training in family planning may be particularly deficient. Residency programs must seek effective teaching strategies to address this deficiency. PMID- 8239852 TI - An epidemic of avian and human psittacosis. AB - Psittacosis may occur in epidemic or sporadic form. We report an outbreak of avian and human psittacosis associated with an aviary in Philadelphia, Pa. The epidemic spread among birds in the aviary and then to men and women who were in contact with the birds. Chlamydia psittaci was cultured from infected birds, and the human patients were evaluated clinically and serologically. After proper treatment of the birds, cleaning of their cages, and institution of appropriate bird-handling techniques, the outbreak among the birds resolved. The human patients were treated, and no subsequent cases have been seen. PMID- 8239853 TI - Hot tub legionellosis. Legionnaires' disease and Pontiac fever after a point source exposure to Legionella pneumophila. AB - Legionella pneumophila is associated with outbreaks of either Pontiac fever, a self-limited influenzalike condition without pneumonia, or Legionnaires' disease, a severe pneumonic disease affecting elderly or immunocompromised individuals. An outbreak of both Legionnaires' disease and Pontiac fever after a point-source exposure to L pneumophila was studied. Our observations demonstrated the spectrum of illness that L pneumophila may cause and emphasized the importance of host factors in affecting the expression of infection. PMID- 8239854 TI - Cardiac rehabilitation and changes in lipid values. PMID- 8239855 TI - Identification of hematopoietic progenitor cells in human amniotic fluid before the 12th week of gestation. AB - Due to technical difficulties in performing amniocentesis before the 12th week of pregnancy, very little is known about the components of human amniotic fluid before that time. Amniocentesis was performed between the 7th-11th week of gestation in 25 informed and consenting patients, who had to undergo therapeutic interruption of pregnancy. The cells from the amniotic fluid were stained, counted on a hemocytometer, and checked for vitality. The origin of these cells was determined from cellular cultures the intracellular content of the hemoglobin and the chloroacetate esterase content of the hemoglobin and the chloroacetate esterase contents were also studied. Taking advantage of the "in vitro" adhesive properties of these cells to the bone marrow stromal feeder layer, we obtained clonal growth of an erythroid nature from 18 out of 33 samples. At the 17th week of gestation, an increased number of cells and a decrease in their vitality was found. Between the 7th and 12th week, the cellular composition of the fluid was totally different from that found later in the pregnancy. Small nucleated, round cells were identified as hematopoietic progenitor cells. At the beginning of the 11th week, a cellular population, typically used to perform prenatal diagnosis of chromosomal abnormalities from the 14th week of pregnancy appeared. Since hematopoietic progenitor cells were found in the amniotic fluid before the 12th week of gestation, these cells most probably come from the hematopoietic cells of the yolk sac through the thin membrane of the yolk sac at this time. PMID- 8239856 TI - The human gonocytes in the early phases of gonadal differentiation. Observations of one embryo at Carnegie stage 11. AB - With the aim of emphasize the characteristics of the migration of human primordial gonocytes through the mesenchyme of the dorsal mesentery towards the genital ridge, a very early human embryo, corresponding to Carnegie stage 11 (23 24 days of development) was studied together with other embryos from the 4th to the 10th week of pregnancy. The early embryo was embedded in resin, serially cut and the observations were made on semithin coronal sections, de-plasticized and stained with toluidine blue. Migrating primordial germ cells were identified in the thickness of the splanchnopleure, where they appeared as pale and round elements with a nucleolated nucleus and granular cytoplasm. The clusters of primordial gonocytes were constantly surrounded and in close relationship with smaller, polyhedral and intensely stained elements. These features suggest the hypothesis that gonocytes and dark cells join together during the process of migration and that the last can participate in the formation of the gonadal bud. The hypothesis is confirmed by the observation of dark cells, surrounding the gonocytes, in the primitive ovaries or testes, in later stages of development. Perhaps they do not participate directly in the formation of the sexual cords, but reach their periphery where they assume an elongate shape. PMID- 8239857 TI - Growth of the body in early fetuses studied by multivariate analysis. AB - The growth of the body was studied in 30 human fetuses ranged from 10 to 22 weeks of gestation. The fetuses were fixed by immersion in 4 percent formaldehyde and the following dimensions were studied: a) lengths: arm, forearm, hand, thigh, leg, foot and crown-rump (sitting height), b) perimeters: head, thorax and abdomen. A covariance matrix was calculated from natural logarithms of all measurements. The relative growth of these measurement was computed by multivariate allometry using a principal components analysis (PCA). All characters were positively correlated with the first principal component which accounted for 94.65 per cent of the total variance. Considering the different measurements in the sequence of the increasing growth rates no one was considered to increase in isometric relationship. PCA showed that the following measurements grew with negative allometry: head perimeter, C-R length, thoracic perimeter, length of the forearm and abdominal perimeter. On the other hand, the following lengths grew with positive allometry: hand, foot, thigh, arm and leg. In conclusion, during the first two trimesters of prenatal life the growth of the body is allometrical. Limbs increase with greater growth rates than the perimeters of the body cavities. PMID- 8239858 TI - The measurement of the calibers of the branches of the aortic arch: a statistical investigation of 430 living subjects using ultrasonic tomography. AB - Using Doppler ultrasound, we have measured the calibers of the branch arteries of the aortic arch in 430 subjects (230 men and 200 women; age range: 18 to 84 years). These included: the brachiocephalic trunk, the subclavian arteries, the common carotid arteries, the internal and external carotid arteries, and the vertebral arteries. Statistical analysis of the values obtained yielded a number of interesting results. The average caliber of the arteries of the women examined was lower than that of the men. Nonetheless, the statistical difference in the arterial diameters of the two sexes was not significant, with the exception of the left subclavian artery. In the case of this artery, its caliber was larger among the men than the women. However, this relationship was not confirmed in the right subclavian artery. The left vertebral artery was larger than the right one. This difference was statistically significant among the women (p < 0.05), but not among the men (p = ns). The right subclavian artery was larger than the left one by a statistically significant margin (p < 0.05), but the statistical significance may be accounted for by the values in the women alone (p < 0.02). When the men were considered separately, the difference between right and left did not achieve statistical significance (p = ns). There was no statistically significant correlation between arterial caliber and either height or body weight. Furthermore, the caliber of the arteries examined increased with age, but not in a statistically significant manner. The mean values of the single arteries examined were lower than those described by the principal investigators of the cadaveric studies to which we have referred. However, with three exceptions, the differences were not statistically significant. In the case of the subclavian and internal and external carotid arteries, our values were lower by a statistically significant margin. PMID- 8239859 TI - Atrophy of the soleus muscle in the albino rat induced by immobilization. AB - The morphological changes in the soleus muscle, immobilized by means of a plaster cast applied to the posterior limb of albino rats was studied. The animals were sacrificed 20 and 30 days after immobilization. The Authors agree with the hypothesis put forth by Lazarides regarding the cytoskeleton model of striated muscle fiber: morphological changes of the sarcomeres and myofilaments are brought on by translateral and longitudinal bridge damage. PMID- 8239860 TI - Cementation for the giant cell tumor of bone. PMID- 8239861 TI - [Skeletal muscle pump as an alternative to heart transplantation]. PMID- 8239862 TI - Effect of total parenteral nutrition on liver mitochondrial function in mature rats. AB - To evaluate the effects of TPN on the hepatic function, the changes in hepatic energy charge levels, oxidative and phosphorylative activities of mitochondria and serum transaminase were studied, using male Sprague-Dawley rats 240 to 250 g in weight. The rats were randomized into three groups. The first group (TPN-V group, n = 6) was infused with TPN solution via the right jugular vein. The number of calories of TPN solution infused daily was adjusted to provide each rat with 80 kcal/kg/day on the 1st day, 160 kcal/kg/day on the 2nd day and 240 kcal/kg/day on the 3rd day. After the 4th day, 240 kcal/kg/day was given to both groups. The second group (TPN-G group, n = 5) was infused with the same solution via an intragastric route and was given the same calories as the TPN-V group. The third group (control group, n = 6) was given a chow diet with the same calories as the TPN group. At the 13th day, all groups were sacrificed, and the hepatic energy charge (EC) and phosphorylation rate (PR) of hepatic mitochondria were measured, and liver function tests were done. PR was 101.2 +/- 5.0 nmol/mg protein/min in control group, 120.8 +/- 2.7 in TNP-G group and 136.5 +/- 6.2 in TPN-V group. EC was 0.906 +/- 0.006, 0.889 +/- 0.008, 0.831 +/- 0.010, respectively. The liver function tests of all group were normal. In both TPN groups, despite evidence that liver function tests were normal, enhanced mitochondrial phosphorylative activity was observed during the early stage of TPN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239863 TI - Isolated perfusion of rat livers: effect of temperature on O2 consumption, enzyme release, energy store, and morphology. AB - The effect of temperature on isolated rat liver perfusion was studied. Livers were perfused for 12 hours with oxygenated Krebs-Henseleit solution at 5, 10, 15, 20, 25 and 30 degrees C, followed by one hour normothermic reperfusion. After each perfusion, oxygen consumption, liver enzyme release, tissue swelling, energy metabolism and histopathological abnormalities were determined. Compared to the oxygen consumption at 37 degrees C, that of 25, 10 and 5 degrees C was 47%, 16% and 12%, respectively. When the liver was perfused at 30 degrees C, higher enzyme release and lower energy status was observed. Tissue swelling was significant only with livers perfused at 5, 10 and 30 degrees C. After normothermic reperfusion, liver injury indicated by enzyme release and bile production was remarkable with 30 degrees C liver, and that of the other groups was essentially the same as the control. ATP of 5, 25 and 30 degrees C liver was significantly lower than the control. Histopathological examination demonstrated abnormalities of sinusoidal cells and hepatocytes in livers perfused at 25 degrees C and 30 degrees C Thus higher temperature (25 degrees C to 30 degrees C) during continuous perfusion were found to induce liver damage. Moderate hypothermia between 10 degrees C and 20 degrees C maintained structure and function of rat livers rather well. These results suggest that, when a machine perfusion of the liver is attempted, a wider temperature range, higher than the conventional, needs to be taken into consideration. PMID- 8239864 TI - [Induction of HSP70 and neuronal damage following transient cerebral ischemia in rats]. AB - Various studies have demonstrated increased synthesis of heat shock protein 70 (HSP70) in brain following transient ischemia, and a protective role for HSP70 against ischemic insult has been hypothesized. In this study, we determined the time course of HSP70 mRNA and HSP70 induction in rat hippocampus following ischemia using Pulsinelli's four-vessel occlusion model, and suggested a protective role for HSP70 induction in limiting ischemic damage to neurons and delayed neuronal death. In Northern blotting analysis using human HSP70 DNA (pH 2.3) as a probe, the accumulation of HSP70 mRNA became evident at 4 h, and continued until 16 h, after 5 min ischemia, while it appeared at 2 h, and continued above control level until 24 h, after 30 min ischemia. In immunoblot analysis using anti-HSP70 antibody, induction of HSP70 appeared 24 h and reached a maximum level 48 h after 5 min ischemia. In immunohistochemical analysis using anti-HSP70 antibody, no staining was detected until 16 h after 5 min ischemia but staining in CA1 gradually increased from 1 day after ischemia and reached a maximum level 2 days after ischemia. Similar time profiles in staining pattern of HSP70 were observed in CA3 and CA4 neuronal cells following 30 min ischemia. Rats pretreated with 5 min ischemia (nonlethal for CA1 pyramidal neurons) were exposed to a 30 min, lethal period of ischemia, 2 days after pretreatment, at which time considerable staining of HSP70 was present. Pretreated rats had much neuronal damage in the CA1 sector less than did rats subjected to lethal, 30 min ischemia alone. These results suggest that neurons in rat hippocampus become tolerant to lethal treatment due to expression of the HSP70 gene and HSP70 protein synthesis induced by mild ischemic pretreatment. PMID- 8239865 TI - [Experimental analysis of cytotoxicity mediated by activated macrophages against glioma cells in mice]. AB - Macrophage (M phi) has a very important role in host natural defence and tumor cell killing. Activated M phi with tumor cytotoxicity can be induced by various lymphokines, such as interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, IL-4, granulocyte macrophage-colony stimulating factor (GM CSF), and macrophage-colony stimulating factor (M-CSF), but also by various bacterial products, such as lipopolysaccharide (LPS), Bacillus Calmette Guerin (BCG), corynebacterium parvum (C.p.), and muramyl-dipeptide (MDP). Although the brain is considered as an "immunologically privileged site", it has been demonstrated that in malignant brain tumors infiltration of lymphocytes or M phi can be seen in tissue samples. This may suggest that immune surveillance exists in the brain. But the biological significance of infiltrative M phi is unclear. The purpose of this study is to confirm the significant generation of activated M phi with cytotoxicity to glial tumor cells and to elucidate the cytotoxic mechanism associated with cellular interaction between effector M phi and target cells. This is the first report on experimental analysis of cytotoxicity mediated by activated M phi against glioma cells in mice. The cytotoxicity mediated by murine peritoneal activated M phi was examined against 3 kinds of murine glioma cell lines; VM-Glioma (spontaneously occurring astrocytoma of the VM mouse origin, H-2b), RSV-M (Schmitt-Ruppin Rous sarcoma virus-induced malignant glioma of the C3H/He mouse origin, H-2k), and 203-Glioma (methylcholanthrene-induced ependymoblastoma of the C57BL/6 mouse origin, H-2b). Activated M phi were obtained from peritoneal exudate cells of 4 strains of mice, C57BL/6 (H-2b), C3H/He (H-2k), DBA/2 (H-2d), and BALB/c (H-2d), following intraperitoneal injection of (1) LPS 200 micrograms, (2) BCG 200 micrograms, (3) C.p. 200 micrograms, (4) MDP 350 micrograms, and (5) IFN-gamma 10(3) units, 7 days prior to 20 hr 51Cr release-cytotoxicity assay. Of the various combination of mouse strains and activating agents tested, that of activated M phi of the C3H/He mouse with induction by LPS had the most tumoricidal effect against the glioma cells, which was not MHC restricted. Although LPS-activated M phi underwent marked loss of cytotoxicity within 24 hr following initiation of in vitro culture, this 20 hr pretreatment with IFN-gamma or TNF-alpha inhibited this reduction in tumoricidal effects in a dose dependent fashion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239866 TI - [Unexpected sudden death and coronary lesions. Apropos of 407 cases out of 1000 deaths in patients under 65 years of age]. AB - In order to determine the responsibility of coronary artery disease in unexpected sudden death (without known or treated cardiac disease), the authors studied 1,000 cases of "natural" unexpected sudden death in subjects under 65 years old in whom the autopsies were performed in Medico-Legal Institutes and in whom all non-cardiac causes had been excluded. Macro- and microscopic examination of the heart detected 848 potentially lethal lesions (152 autopsies failed to show the cause of death) of which 407 cases corresponded to coronary disease (atherosclerosis in 340 and other disease in 67 cases). The atherosclerotic lesions could be classified in 3 anatomical groups: 75 isolated recent coronary thromboses; 75 recent coronary thromboses associated with multivessel stenoses; 190 coronary stenoses (> 75%) without thrombosis (139 triple vessel, 31 double vessel and 20 single vessel diseases). The presence of a recent myocardial infarct, the cardiac mass and the circumstances of sudden death were determined in all cases with respect to the three pre-defined anatomical groups. This study, though confirming the high prevalence of coronary artery disease, evaluates it at a lower level than previous reports in the literature which takes into consideration "unexpected sudden death" of known and treated coronary patients, and also has allowed identification of other potentially lethal lesions (such as structural abnormalities of the His-Purkinje system) which are undetectable even with sophisticated paraclinical investigations. PMID- 8239867 TI - [Short- and mid-term results of percutaneous mitral valve commissurotomy for restenosis after surgical commissurotomy]. AB - Fifty nine of the 280 percutaneous mitral valvuloplasties (21%) performed between March 1987 and December 1991 at the Montreal Heart Institute were carried out for symptomatic mitral restenosis 15 +/- 6 years after surgical commissurotomy. The patients were selected according to echocardiographic criteria. The mitral valve disease was comparable to that of patients without previous surgical commissurotomy. Patients with good hemodynamic result had undergone surgical commissurotomy more recently and had less severe valvular damage than patients with an incomplete (n = 10) or poor result (n = 7). One patient died of a cerebral embolism during the procedure, two patients underwent emergency surgery for a mitral valve rupture and 4 patients had an atrial septal defect with Qp/Qs ratio greater than 1.5 by oximetry. The numbers of successes and complications were comparable to those observed in patients without previous surgical commissurotomy. The patients who had undergone previous surgical commissurotomy were followed up for 1 year. At 12 months, 4 remained in Class II of the NYHA classification, 44 (74%) were improved by at least 1 functional class and 10 (17%) had required a mitral valve replacement. Percutaneous mitral valvuloplasty is therefore a useful procedure in patients with moderate degrees of valvular disease who develop restenosis after surgical commissurotomy. A functional improvement can be expected in 3/4 of patients Complications related to the procedure are rare and acceptable compared with the risks of a second thoracotomy. Failure of percutaneous mitral valvuloplasty is generally due to the degree of valvular disease which contraindicated surgical commissurotomy and required mitral valve replacement. PMID- 8239868 TI - [Coronary disease with normal coronarography in the black Africans: epidemiological and clinical data in 31 cases. Role of abnormal hemoglobins]. AB - The authors studied 31 cases of coronary artery disease with normal or minimally diseases coronary arteries in black Africans, 29.8% of 104 coronary patients undergoing coronary angiography in this series. These 31 cases comprised 16 cases of infarction, 10 cases of angina, 3 ventricular aneurysms and 2 cases of silent ischemia in diabetic patients. Twenty-five patients were men (80.6%). There were 6 women (19.3%) two of whom presented in the post-partum period. The average age of these patients was 45 years (males: 47.7 years; females: 41.8 years). The following risk factors were noted: smoking (60%), hypertension (25.8%), obesity (29%), diabetes (12.9%), serum cholesterol (average 2.15 g/l), serum triglycerides (average 1.25 g/l). The risk index per patient was 1.29. In comparison with coronary patients with angiographic coronary lesions (n = 73), the patients with normal angiography were significantly younger, comprised more females and had fewer risk factors (especially hypertension and diabetes), though this was not statistically significant. The prevalence of inaugural infarction was 81.2% in the cases of infarction with normal coronary arteries. These infarcts may be complicated by ventricular aneurysm formation. Spontaneous spasm was observed in 3 out of 31 patients (9.6%) at coronary angiography. A provocative test was performed in only 2 cases and 1 was positive. This deserves further study and may have therapeutic implications. The authors emphasise the high incidence of hemoglobin S or C traits (57.1%). These heterozygotic hemoglobinopathies could be a risk factor in these coronary patients with normal coronary angiography. PMID- 8239869 TI - [Value and indication of the use of 2 internal mammary arteries in repeated coronary surgery]. AB - Forty-nine patients who had coronary artery reoperations were divided into two groups: the 29 patients of the first group were operated conventionally with use of one internal mammary artery or a saphenous vein; the 20 patients of the second group were reoperated using both internal mammary arteries. Three patients (6%) died prematurely: two in the first and one in the second group. The rates of peri operative infarction were 7% and 15% respectively. The average postoperative bleeding was 472 +/- 385 ml in the first group and 700 +/- 628 ml in the second group (NS). All patients are pauci-symptomatic and have a negative exercise stress test. The mortality and morbidity of coronary reoperation does not seem to be greater with double internal mammary artery bypass grafting. However, this technique should be reserved for patients who can derive long-term benefit from reoperation with arterial grafts, that is to say in patients in good clinical condition, less than 65 years of age with good left ventricular function. In these patients, double internal mammary artery bypass grafting may avoid a third operation for myocardial revascularisation. PMID- 8239870 TI - ["White-coat" effect in 35 consecutive patients with hypertension. Quantification method, role of physician-patient dialogue]. AB - The "white coat" effect, an alarm reaction to the presence of a doctor, is an important cause of blood pressure variability, the frequency, amplitude and mechanisms of which are only partially understood. In order to evaluate these factors, a prospective study was undertaken in 35 consecutive patients referred for assessment of clinical hypertension. The alarm reaction was investigated during the consultation, at the time of interrogation, in periods of silence, in the sitting and upright positions. Twenty-four to forty-eight measurements (average 36.8) of the blood pressure and heart rate were performed in each patient with a Diasys 200 R monitor. The ambulatory period of 3 to 5 hours after the consultation provided 12 to 24 measurements (average 20.7) which were considered to be the reference for comparison with the consultation period. A total of 2,038 measurement were made and analysed. Analysis of variance (GLM) for each patient and for the whole group gave an assessment of the alarm reaction during the patient-doctor dialogue and periods of silence with reference to the ambulatory period. During the ambulatory period, the average and standard deviation for systolic pressure were 134 +/- 0.7 mmHg, and for diastolic pressure 93.1 +/- 0.6 mmHg. These pressures were significantly lower than during the two periods of consultation, with and without dialogue (p < 0.0001). During the consultation, the systolic and diastolic blood pressure values were significantly higher during the dialogue than during the periods of silence (p < 0.0001). During the dialogue, the systolic pressure attained 153.7 +/- 0.7 mmHg and the diastolic pressure: 107.2 +/- 0.6 mmHg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239871 TI - [Atrioventricular block induced by radiofrequency: electrophysiological criteria of success]. AB - In a retrospective series of 18 consecutive patients (10 men, average age: 61.6 +/- 9.2 years) who underwent His bundle ablation by radiofrequency current, the authors analyzed the electrophysiological criteria predictive of complete atrioventricular block. A total of 82 radiofrequency bursts were analyzed (average: 4.5 +/- 3.1 bursts/patient) and classified as effective (N = 14, definitive atrioventricular block), ineffective (N = 55, no effect on atrioventricular conduction) or transiently effective procedures (N = 13, reversible Mobitz II atrio-ventricular block). Seven electrophysiological parameters were measured: HV interval, amplitude of atrial (A), Hisian (H) and ventricular (V) potentials, and the A/H, A/V, and V/H ratios before each radiofrequency burst. At the end of the ablation, 14 patients were in complete atrioventricular block and during the following 24 hours, a complete atrioventricular block developed in 3 other patients. The average value of the H potential was comparable in the effective and transiently effective procedures (0.28 +/- 0.20 mV and 0.27 +/- 0.19 mV respectively) and significantly higher than the same parameters in the ineffective group of procedures (0.65 +/- 0.14 mV, p = 0.008). The V/H ratio was progressively greater in effective (5.43 +/- 2.51), transiently effective (8.07 +/- 6.90) and ineffective procedure (14.32 +/- 13.35), p = 0.02; the average value of the A amplitude tended to be higher in the effective procedures (1.03 +/- 0.75 mV) and the transiently effective procedures (0.98 +/- 0.72 mV) than in the ineffective procedures (0.58 +/- 0.79 mV), p = 0.06. On the other hand, the other parameters were comparable in the three groups of patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239872 TI - [Results of high amplification electrocardiogram in primary dilated cardiomyopathy]. AB - Idiopathic dilated cardiomyopathy carries a high risk of sudden death. It is also associated with sustained ventricular tachycardia. A complex ventricular arrhythmia is recorded in 3/4 of cases on Holter monitoring which has a low specificity. The aim of the study was to determine whether signal-averaged electrocardiography could provide a better evaluation of the prognosis of this condition. The results of signal-averaged electrocardiography were compared with those of 24 hour Holter monitoring and of systematic programmed ventricular stimulation in 58 patients with idiopathic dilated cardiomyopathy. Late ventricular potentials were recorded in 13 of the 14 subjects with inducible and usually spontaneous sustained ventricular tachycardia. The sensitivity of the technique for evaluating the risk of sustained VT was therefore good (93%). Late potentials were also recorded in 9 patients with induced ventricular flutter or fibrillation, these patients being symptomatic (dizzy spells). Late potentials were also demonstrated in 14 of the 35 asymptomatic patients without inducible VT, indicating that this non-invasive investigation had a limited specificity (60%). In addition, during follow-up of the patients, the risk of sudden death was difficult to demonstrate. Late potentials were only found in subjects with inducible sustained VT but no in the other cases. In conclusion, signal-averaged electrocardiography seems to be valuable for evaluating the risk of sustained VT in subjects with idiopathic dilated cardiomyopathy and complex ventricular arrhythmias. The detection of the risk of sudden death is probably impossible by this technique. PMID- 8239873 TI - [Pulmonary perfusion tomoscintigraphy in pulmonary embolism. Preliminary study]. AB - Tomoscintigraphy, a method developed over 10 years ago, is little used nowadays for the evaluation of suspected pulmonary embolism. The authors report the results of a preliminary study of 10 patients hospitalised for this condition. Tomoscintigraphy was normal in 2 cases and abnormal in 8 cases. Seven of these 8 patients underwent pulmonary angiography which confirmed the diagnosis of pulmonary embolism in 6 cases. The small number of patients, however, did not allow measurement of the sensitivity and specificity. Tomoscintigraphy, repeated at the 8th day and at the first month, provides an assessment of therapeutic efficacy. Some improvement is observed in all cases from the first control but the amount varies from one subject to another. At one month, 3 of the 6 patients undergoing control tomoscintigraphy had hypoperfusion sequellae. The simplicity of pulmonary tomoscintigraphy makes it a useful investigation for emergency diagnosis and follow-up pulmonary embolism. PMID- 8239874 TI - [Peripheral arterial angioplasty: value of the popliteal approach. Apropos of 30 cases]. AB - The percutaneous common femoral arterial approach is usually used for endovascular management of lower limb arterial disease. This approach is sometimes impracticable because the femoral artery is the site of severe calcific atheromatous lesions which prevent arterial puncture or, when the superficial femoral lesions are ostial or proximal, make it impossible to position the introducer and advance the guide wire. The popliteal artery then becomes very useful for treating these lesions by a retrograde approach. Similarly, superficial femoral lesions which cannot be successfully dilated by the anterograde femoral approach may justify retrograde catheterisation via the popliteal artery. Between May 1988 and August 1991, the authors used the retrograde popliteal approach in 30 cases. They obtained 24 successes, 12 of which were associated with the implantation of an endoprosthesis. There was 1 complication at the puncture site a popliteal arteriovenous fistula was created but was treated successfully by surgery. PMID- 8239875 TI - [Left intraventricular dynamic gradients in the follow-up of aortic valve replacement: an echo-Doppler study]. AB - The possibility of an intraventricular pressure gradient in patients with aortic stenosis is well known: this entity is associated with a high risk of postoperative complications. The authors carried out a Doppler echocardiographic study of flow in the left ventricle in 51 patients who had recently undergone valve replacement for severe aortic stenosis (valve area < 0.75 cm2). Before surgery, only one patient had significant acceleration of intraventricular systolic flow attaining 3.8 m/s (maximum pressure gradient of 60 mmHg). After surgery, maximum intraventricular systolic velocities of over 2.5 m/s with a typical end systolic peak were observed in 8 patients under basal conditions (gradients of 30 to 115 mmHg), and in 7 others after inhalation of amyl nitrite. Pulsed spectral and color Doppler flow mapping showed that the highest velocities were located at the mitral papillary muscle level. In addition, these patients had significant reduction in cavity size. Only one patient had systolic anterior motion of the anterior mitral leaflet with septal contact. Left ventricular dimensions were measured by TM echocardiography. High intraventricular velocities seemed to be significantly related to the smallest ventricular dimensions, the thickest ventricular walls and the smallest preoperative aortic valve surface area. The highest intraventricular pressure gradients-disappeared with betablocker therapy (4 cases), after correction of hypovolemia (1 case), after drainage of large pericardial effusions (2 cases) or spontaneously (1 case). This study confirms the relatively high prevalence of dynamic intraventricular gradients after surgical cure of aortic stenosis and the value of Doppler echocardiography for the avoidance of certain drugs (inotropic agents, vasodilators, diuretics), which could aggravate the hemodynamic abnormality.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239876 TI - [Changes in plasma atrial natriuretic factor, plasma and urinary cyclic GMP during exercise in coronary patients and healthy subjects]. AB - The concentrations of plasma ANF and plasma and urinary cyclic GMP were measured at rest and during exercise in 12 normal subjects (reference group) and 20 patients with coronary artery disease (coronary group). In both groups, plasma ANF and c GMP increased during exercise and fell one hour after (F = 3.8, p = 0.029 and F = 13.3, p = 0.0001, respectively) whereas the urinary c GMP increased one hour after exercise (F = 5.3, p = 0.029). In the control group, ANF increased on effort and fell during recovery to above its resting value whereas the plasma c GMP remained unchanged throughout the test. In the coronary group, no significant increase in ANF was observed on effort (wide dispersion of values) whereas the c GMP increased during effort and fell to below testing value during the recovery phase. The ANF of the coronary group was globally higher than the ANF of the control group (F = 4.7, p = 0.04). The plasma c GMP of the coronary group was comparable to that of the controls (F = 2.1, p = 0.15) despite higher concentrations at rest (p < 0.05) and during exercise (p < 0.05). However, there was a positive interaction between the efforts of exercise and the pressure of coronary disease on the concentration of plasma c GMP (F = 6.7, p = 0.0024). There was no difference in urinary c GMP between control and coronary subjects (F = 1, p = 0.33).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239877 TI - [Guidelines for the use of implantable defibrillators. A "Task Force" of the working groups in cardiac arrhythmia and cardiac stimulation of the European Society of Cardiology]. PMID- 8239878 TI - [Myocardial infarction and effort rhabdomyolysis. Problems caused by serum enzymatic assays]. AB - The authors report the case of a patient who presented with myocardial infarction and rhabdomyolysis. The mild ECG changes in the basal leads and the normal MB creatinine phosphokinase fraction, expressed as a percentage of total CPK, delayed the diagnosis of myocardial infarction. The presence of a rhabdomyolysis without any clinical manifestation was suspected from the severe and prolonged increase in cardiac enzyme levels. Contrary to the usual method of measuring the MB fraction of CPK by immuno-inhibition, the fluorimetric immuno-enzymological technique enabled diagnosis of the infero-lateral-basal infarct, later confirmed by echocardiography and left ventriculography. This new technique would seem to be very useful, given the high sensitivity and specificity and its rapidity not only in the context of myocardial infarction associated with rhabdomyolysis, but also in the frontier forms of unstable angina and definitive myocardial infarction. PMID- 8239879 TI - [Incidental finding of an isolated thrombus in the left atrium in a young woman with sinus rhythm 5 years after cerebral ischemic accident]. AB - The authors report the case of a 40 year old woman who had an ischemic stroke. The initial investigation including a complete blood clotting analysis, failed to demonstrate the cause. Five years later, the investigations were completed systematically by transesophageal echocardiography which demonstrated an isolated thrombus localised in the left atrial appendage though the localisation heart was in sinus rhythm and morphologically normal. This case illustrates in the diagnostic yield of transesophageal echocardiography in the investigation of systemic embolism in young patients who have had a cerebral ischemic event of unknown cause, even in the absence of predisposing cardiac cause. PMID- 8239880 TI - The biological role of nitric oxide in bacteria. PMID- 8239881 TI - Effects of dilution rate and pH on the ruminal cellulolytic bacterium Fibrobacter succinogenes S85 in cellulose-fed continuous culture. AB - The ruminal cellulolytic bacterium Fibrobacter succinogenes S85 was grown in cellulose-fed continuous culture at 22 different combinations of dilution rate (D, 0.014-0.076 h-1) and extracellular pH (6.11-6.84). Effects of pH and D on the fermentation were determined by subjecting data on cellulose consumption, cell yield, product yield (succinate, acetate, formate), and soluble sugar concentration to response surface analysis. The extent of cellulose conversion decreased with increasing D. First-order rate constants at rapid growth rates were estimated as 0.07-0.11 h-1, and decreased with decreasing pH. Apparent decreases in the rate constant with increasing D was not due to inadequate mixing or preferential utilization of the more amorphous regions of the cellulose. Significant quantities of soluble sugars (0.04-0.18 g/l, primarily glucose) were detected in all cultures, suggesting that glucose uptake was rather inefficient. Cell yields (0.11-0.24 g cells/g cellulose consumed) increased with increasing D. Pirt plots of the predicted yield data were used to determine that maintenance coefficient (0.04-0.06 g cellulose/g cells.h) and true growth yield (0.23-0.25 g cells/g cellulose consumed) varied slightly with pH. Yields of succinate, the major fermentation endproduct, were as high as 1.15 mol/mol anhydroglucose fermented, and were slightly affected by dilution rate but were not affected by pH. Comparison of the fermentation data with that of other ruminal cellulolytic bacteria indicates that F. succinogenes S85 is capable of rapid hydrolysis of crystalline cellulose and efficient growth, despite a lower mu max on microcrystalline cellulose. PMID- 8239882 TI - A new structural type of teichoic acid and some chemotaxonomic criteria of two species Nocardiopsis dassonvillei and Nocardiopsis antarcticus. AB - The cell-wall teichoic acids of Nocardiopsis dassonvillei IMRU 509T, IMRU 504 and IMRU 1250 and Nocardiopsis antarcticus VKM Ac-836T have the same unique structure that has not heretofore been found in bacteria. The polymer is built of 10 to 13 repeating units: -1snGro3-P-3-beta-D-GalNAc1-2snGro3-P-. The structure has been established by chemical analysis and with NMR spectroscopy. A study of the cell wall monosaccharide composition, peptidoglycan, and cell phospholipids also provides evidence for the close similarity of these two species. PMID- 8239883 TI - Induction of pyruvate decarboxylase in glycolysis mutants of Saccharomyces cerevisiae correlates with the concentrations of three-carbon glycolytic metabolites. AB - Pyruvate decarboxylase, PDCase, activity in wild-type yeast cells growing on ethanol is quite low but increases up to tenfold upon addition of glucose, less with galactose and only slightly with glycerol. PDCase levels in glycolysis mutant strains growing on ethanol or acetate were higher than in the wild-type strain. These levels correlated with the sum of the concentrations of three carbon glycolytic metabolites. The highest accumulation was observed in a fructose bisphosphate aldolase deletion mutant concomitant with the highest PDCase activity ever observed under gluconeogenic conditions. Glucose addition induced an increase in PDCase activity in all mutants. However, the enzyme activities never reached wild-type level. On the other hand, the PDCase levels in the different mutants again correlated with the sum of the concentrations of the three-carbon glycolytic metabolites. This was interpreted to mean that full induction of PDCase activity requires the accumulation of hexose- and triosephosphates. PMID- 8239884 TI - Analysis of Candida utilis genomic DNA, homologous to cDNA of chicken A1(1) collagen gene. AB - Genomic fragments, homologous to chicken A1(1) collagen cDNA encoding triple helical domain, were revealed by Southern analysis in various fungi. Such a genomic fragment from Candida utilis was cloned and sequenced. Analysis of the obtained DNA sequence revealed the 119 bp segment, which has possibly originated from the 54 bp module common for the fibrillar collagen genes of higher eukaryotes. PMID- 8239885 TI - [Treatment of children with short stature with human growth hormone and its potential risks]. PMID- 8239886 TI - [Europe, pediatrics and pediatricians]. PMID- 8239887 TI - [What about the ambulatory pediatrics?]. PMID- 8239888 TI - [Criteria for classification of spondylarthropathies in children]. AB - BACKGROUND: The term spondylarthropathy is used for several different entities including undifferentiated forms. The latter are particularly frequent in children and often not recognized. Two sets of criteria, the Amor and the European Spondylarthropathy Study Group (ESSG) criteria, primarily devised for adults, were evaluated in a pediatric population. METHODS: Three hundred and ten consecutive patients referred to one pediatric rheumatology center over 6 months were prospectively assessed, whatever the rheumatic complaint. Twenty two items were analysed for each patient and compared with the criteria proposed by Amor (17 items) and the ESSG (12 items). RESULTS: The Amor criteria performed better than the ESSG criteria. The sensitivity was 84 and 69.7%, and the specificity 96.1 and 92.2% for the Amor and the ESSG criteria, respectively. The sensitivity for pauci-arthritis was much higher in children, but the specificity was lower than in adults. The sensitivity of spinal involvement was very low in children. The undifferentiated type accounted for 81% of all definite spondylarthropathies. Definite and possible spondylarthropathies were 17% of the total referral over the 6 months, i.e. one patient in 6. CONCLUSIONS: These two sets of criteria, particularly the Amor criteria, can help pediatricians to recognize the undifferentiated type of spondylarthropathy, in the absence of spinal or any other characteristic manifestation. PMID- 8239889 TI - [Isolated acute peripheral facial paralysis in children. Etiological and prognostic study]. AB - BACKGROUND: As facial nerve palsy is usually of sudden onset and of obscure etiology, a survey of large series may provide new information. PATIENTS AND METHODS: The files of 40 children suffering from acute facial nerve palsy seen in 3 pediatric and 1 otorhinolaryngologic (ORL) departments between 1980 and 1990 were analysed. Patients with congenital palsy and those with paralysis that appeared after injury or in the course of acute or chronic disease (otitis media, tumor) were excluded from the study. The children were 9 months-16 years old (mean: 7 year 1 month). There were 22 left and 18 right facial nerve palsies. The palsy was moderate in 17 children, severe in 15, and total in 5. Etiologic factors (viral serologic studies, CSF examination), ORL studies and electromyogram of the facial nerve were analyzed in the majority of cases. Corticosteroids were used in 30 patients. RESULTS: A viral etiology was suspected and/or confirmed in 13 children; abnormalities were seen in 6 of the patients who underwent lumbar puncture. A familial facial nerve palsy was seen in 1 case and kidney disease in 2 cases. 23 cases appeared to be idiopathic. A moderate permanent residual weakness was seen in only 4 children. The mean time for recovery was 32 days (4-140 days). Corticosteroids did not influence the incidence of complete recovery, but decreased the time for recovery (26 versus 37 days) only in the patients who were treated before the 3rd day of the disease. Patients with idiopathic palsy recovered 23 days before those with viral etiology. CONCLUSIONS: Idiopathic and post-viral facial nerve palsies seem to have an excellent prognosis. Corticosteroids seem to have no effect although there is some evidence of benefit when treatment is begun early. PMID- 8239890 TI - [Technetium 99m labeled dimercaptosuccinic acid (99m Tc-DMSA) scintigraphy in the diagnosis and follow-up of urinary infections in children]. AB - BACKGROUND: The site of a urinary tract infection (UTI) is located by clinical findings plus imaging techniques. Renal imaging with technetium 99mTc dimercaptosuccinic acid (99mTc-DMSA) has shown promise but its efficacy for detecting acute pyelonephritis is still debated as is its ability to differentiate between acute, potentially curable disease and scarring, definitive changes. PATIENTS AND METHODS: The files of all the 166 patients that underwent one or two (32 patients) 99mTc-DMSA imaging sessions for UTI in 1989 and 1990 in our department were analysed. The results of this technique were compared with the clinical and laboratory data and with those of other imaging techniques (ultrasonography and retrograde voiding cystourethrogram). The patients (92 girls and 74 boys, mean age: 5 years, range 15 days-17 years) were assigned to one of 3 categories: 1) acute pyelonephritis (27 patients), 2) low UTI (60 patients) and 3) uncertain UTI (79 patients), on the basis of the clinical and laboratory data. RESULTS: The first 99mTc-DMSA imaging was normal in 110 children; changes were unilateral in 45 and bilateral in 11 children. Images suggesting either acute or chronic changes were observed respectively in 19 and 48 children. 4 of the 7 acute changes were normalized on the second radioisotope scanning while 2 became worse; 19 of the 23 chronic changes were unchanged at the second scanning. Radioisotope scanning changes were seen in 81% of the patients classified as acute pyelonephritis and in only 27% of those with lower UTI. Reflux was detected in 56% of abnormal kidneys and in 36% of normal kidneys. The results of radioisotope scanning and ultrasound scan findings were not correlated in 24 patients (19 only abnormal radioisotope imaging, 5 only abnormal ultrasound scan). CONCLUSIONS: DMSA scans are useful for investigating and following UTI. They help to distinguish acute pyelonephritis and lower UTI. It is a reliable method of detecting structural abnormalities and identifying children at risk of progressive renal damage. PMID- 8239891 TI - [The effect of human albumin in association with intensive phototherapy in the management of neonatal jaundice]. AB - BACKGROUND: The effectiveness of phototherapy in lowering serum bilirubin levels in neonates varies inversely with the rate and degree of hemolysis. Combining this therapy with albumin perfusion could enhance its effectiveness. This study examines total, unconjugated and unbound fractions of bilirubin in infants treated by phototherapy alone or by phototherapy plus albumin. PATIENTS AND METHODS: The files of 211 neonates treated from January 1990 to March 1991 for severe neonatal jaundice were analysed. Jaundice was due to ABO incompatibility in 113 cases and hereditary hemolytic anemia in 6 cases; its cause was unknown in 92 patients. Other causes of jaundice such as Rh incompatibility, premature delivery before 34 gestational weeks and neonatal infections were excluded from the study. All 211 neonates were given phototherapy from admission with similar light energy. 114 babies (group I) received only phototherapy, while the 97 others (group II) were also given human albumin, (1.5 g/kg), during the first 2 hours of phototherapy. RESULTS: The decrease in serum unconjugated and unbound bilirubin after 4 hours of phototherapy was 34% in group I and 45% in group II (p < 0.0005). There was no difference between both groups after 24 hours of phototherapy. Other factors such as the initial concentration of serum unconjugated and unbound bilirubin, age at the onset of therapy, and ethnic skin color also influenced the decrease in bilirubin after 4 and 24 hours of phototherapy, in addition to albumin perfusion. CONCLUSIONS: Albumin perfusion plus phototherapy appears to induce a rapid and early decrease in unconjugated, unbound bilirubin, the fraction that is potentially neurotoxic, while phototherapy alone acts over a longer period. PMID- 8239892 TI - [Malignant tumors of the cerebral hemisphere in children. A series of 64 cases]. AB - BACKGROUND: Cerebral hemisphere tumors in children represent about 15% of all tumors of central nervous system. 30 to 40% of them are malignant and have a poor prognosis. PATIENTS AND METHODS: The files of 64 children (30 boys and 34 girls, less than 15 years old) treated between 1970 and 1989 for malignant cerebral hemisphere tumor were analysed. There were 27 astrocytomas, 20 ependymomas, 7 oligodendroblastomas, 6 sarcomas, 2 neuroblastomas, 1 medulloepithelioma and 1 teratoma. The tumor was located in the frontal area in 18 patients; it was temporal in 9, parietal in 8 and occipital in 2 patients. The tumor infiltrated 2 or several lobes in 27 children. The tumor was excised in 58 patients (completely in 36). Radiotherapy was given in 42 children; 18 of them also received chemotherapy. RESULTS: The time between the first clinical manifestation and diagnosis was longer than 6 months in 20% of cases. Symptoms of increased intracranial pressure were present at onset of the disease in 56% of patients and at admission in 73%. Convulsions revealed the tumor in only 30% of cases. Nine children died during the first post-operative month. The actuarial survival rate was 39 +/- 13% at 5 years and 29 +/- 13% at 10 years. This rate was better for ependymomas (48 +/- 23% at 5 years and 41 +/- 23% at 10 years) than for astrocytomas (25 +/- 18% and 13 +/- 16%). Three of the 27 astrocytomas developed in children who had been treated 6 to 7 years earlier for acute lymphoblastic leukemia. One infant developed acute lymphoblastic leukemia 2 years after surgery and chemotherapy for astrocytoma and one patient, treated at 5 months of age for retinoblastoma, developed an astrocytoma 13 years later. CONCLUSIONS: Increased intracranial pressure is more frequently the first symptom than are convulsions in malignant cerebral hemisphere tumors. The long interval between the first manifestations and diagnosis suggests degeneration of benign tumors in 20% of cases. The development of a second malignant neoplasm is not exceptional in children. The better prognosis for ependymomas than for astrocytomas indicates that cerebral ependymomas should be excised and irradiated regardless of their grade. PMID- 8239893 TI - [Severe colectasy in rheumatoid purpura. Probable role of water soluble contrast products]. AB - BACKGROUND: About 60-70% of cases of anaphylactoid purpura suffer from abdominal colic, which may be quite severe. Because severe abdominal pain may be difficult to differentiate from a surgical abdomen, repeated examination and radiological studies are often necessary. CASE REPORT: A 4 year 7 month-old girl suffered from abdominal pain, vomiting and the skin rash characteristic of anaphylactoid purpura. Because of worsening of the abdominal manifestations, a Gastrografin enema was given. The result was normal but 48 hours later, the child suffered from acute abdominal colics plus symptoms of shock. X-ray examination revealed a large distension of colon and persistence of the contrast material. Exploratory laparotomy failed to find any perforation or necrosis. The colon was drained and the child was given parenteral nutrition and antibiotics. Blood culture showed Klebsiella pneumoniae. An upper gastrointestinal roentgenogram using meglumine ioxitalamate was performed 19 days later because of recurrence of the abdominal pain. This showed a submucosal hemorrhage in the first small bowel loop. Increased abdominal distension occurred 3 days later and the results of X-rays were similar to those seen the Gastrografin enema. A second enema using meglumine ioxitalamate was performed 3 days later. It showed sigmoid constriction but a second laparotomy failed to confirm this obstruction. The child died a few hours later despite ileostomy and antibiotics. CONCLUSIONS: The severe colectasy seen in this case of anaphylactoid purpura indicates that caution is required in performing enemas in acute digestive complications of this disease. Such X-ray studies should not be repeated and all water-soluble contrast material should be voided as soon as possible. PMID- 8239894 TI - [Atrioventricular block complicating the use of diphemanil (Prantal) in 2 premature newborn infants]. AB - BACKGROUND: Diphemanil can be useful in some neonates presenting with bradycardia due to vagal hyperreflectivity. Paradoxically, this drug may induce atrio ventricular (AV) block in premature babies. CASE REPORTS: Case no 1. A premature neonate suffering from acute respiratory distress from birth required respiratory support, antibiotics and caffeine. Despite this treatment, he underwent many episodes of apnea, and bradycardia that appeared on day 4 and did not respond to IV doxapram (1 mg/kg/h). He was given diphemanil on day 9 (10 mg/kg/d) and permanent bradycardia with complete AV block and a normal QT interval appeared 2 days later. Cessation of diphemanil and administration of IV isoprenaline led to a normal sinusal rhythm, but there were bladder, intestinal and ocular signs of atropinic intoxication. A complete definitive recovery occurred 5 days after cessation of diphemanil. Case no 2. A premature neonate developed episodes of apnea 2 days after birth. These episodes persisted and were complicated by bradycardia on day 4 despite administration of caffeine. Vagal stimulation on day 7 was positive and the infant was then given diphemanil (10 mg/kg/d). Permanent bradycardia occurred 2 days later, with partial AV block and a normal QT interval. The child recovered a normal sinusal rhythm 2 days after cessation of diphemanil. CONCLUSIONS: Anticholinergic therapy may cause permanent bradycardia due to AV block in premature infants. This therapy should not be given to premature infants without a prior ECG. Doses lower than those used in infants are recommended. PMID- 8239895 TI - [Anuria after abdominal surgery in 2 newborn infants. Beneficial effect of noradrenaline]. AB - BACKGROUND: Abdominal surgery in neonates may be complicated by acute renal failure that is sometimes due to increased intra-abdominal pressure. Correction of the decreased renal perfusion may be difficult. CASE REPORTS: Case no 1. A girl weighing 3,000 g was operated on at 3 hours of life for congenital omphalocele. Closing the defect induced increased intra-abdominal pressure and decreased systemic pressure. Despite dopamine, dobutamine, followed by furosemide, anuria persisted without manifestations of heart failure. Noradrenaline (0.1 to 0.7 micrograms/kg/min intravenously) given 24 hours after surgery resulted in normalization of systemic pressure and diuresis. Case no 2. A boy was born at gestational week 30, weighing 1,450 g. At 8 days of age, he was suffering from shock that was attributed to perforation of the bowel due to necrotizing enterocolitis. Surgery was complicated by acute circulatory and renal failure that did not respond to dopamine, dobutamine or furosemide. Infusion of noradrenaline, (0.2 to 0.6 micrograms/kg/min intravenously) induced diuresis within 3 hours. CONCLUSIONS: Noradrenaline can be useful in patients with postoperative increased intra-abdominal pressure. It should only be given after correction of hypovolemia, control of myocardial function, and when dopamine remains ineffective. PMID- 8239896 TI - [Precocious puberty revealing Bourneville tuberous sclerosis]. AB - BACKGROUND: Benign tumors are seen in tuberous sclerosis. They are found in many organs, and the precocious puberty due to hypothalamic hamartoma and tuberous sclerosis has been reported. However, precocious puberty exceptionally reveals the tuberous sclerosis. CASE REPORT: A 2 month-old boy was admitted because of the fortuitous discovery of polycystic renal disease. Precocious puberty developed at 13 months with enlargement of the penis and testes, appearance of pubic hair, acne and deepening of the voice. Linear growth was recently accelerated and the bone maturation was advanced. Plasma testosterone was elevated (460 ng/100 ml) and LH-RH injection induced rapid rises in plasma LH (2.6 to 28 mUl/ml) and FSH (2 to 8 mUl/ml). Brain imaging techniques (CT scan and NMR) showed a hypothalamic hamartoma and periventricular calcified lesions. Examination with the Wood lamp identified two white leaf macules in the dorsal area. Administration of an analogue of LH-RH effectively reduced the manifestations of precocious puberty. CONCLUSION: Tuberous sclerosis is exceptionally revealed by precocious puberty. The association of polycystic disease and precocious puberty has never before been reported in tuberous sclerosis. PMID- 8239897 TI - [Iconographic rubric. Menkes disease revealed by very severe neurologic development]. PMID- 8239898 TI - [Streptococcal B infections in neonatal period. Epidemiology and prevention]. PMID- 8239899 TI - [Septicemia due to Streptococcus C revealing pulmonary tuberculosis]. PMID- 8239900 TI - [Prevalence of skin sensitivity to allergens in asthmatic children in the Paris region]. PMID- 8239901 TI - [Maternal serum markers and prenatal screening for trisomy 21]. PMID- 8239902 TI - Therapeutic potential of biological response modifiers against transplantable mouse tumors of spontaneous origin. I. Characterization of biological properties of tumor lines and preliminary data on response to cytostatic drugs and biomodulators. AB - Several transplantable lines were derived from spontaneous tumors of mouse strains of low and high cancer incidence (CBA, BALB/c and DBA/2), and eight of them were used for an evaluation of the therapeutic potential of BRM. The lowest number of cells inducing tumors was in most cases around 10(4) cells. In transplantation tests, tumors of CBA and DBA/2 mice were non-immunogenic, while tumors of BALB/c mice showed different level of immunogenicity. Transplantation of a small number of tumor cells in admixture with BCG resulted in no tumor growth of the immunogenic line X6 and no effect on take rate of non-immunogenic line X1. Sensitivity of studied tumor lines to BCNU, CY and ADR was moderate. Therapy of the tumors with postulan and CMA, two biomodulators already shown to be active against various experimental tumors, failed. The significant therapeutic response was observed in DBA/2 mice bearing i.p. transplant of lymphosarcoma X19 and treated with i.p. injections of CMA. PMID- 8239903 TI - Synthesis and biological properties of 5-hydroxy-methylpyrimidines. AB - Reduction of 4-arylamine-6-methyl-2-phenyl-5-pyrimidine carboxylic acid and its ethyl esters as well as 5,7-dihydrofuro (3,4-d)-pyrimidines resulted in obtaining some 4-arylamine-6-methyl-2-phenyl-5-hydroxymethylpyrimidines exhibiting strong immunomodulatory and cytostatic properties. PMID- 8239904 TI - Effect of amino acids on the pig muscle enolase. AB - Fifteen amino acids were used to investigate their effects on the pig muscle enolase. The basic and hydrophobic amino acids decreased the enzyme activity slightly. In the presence of acidic amino acids such as aspartic and glutamic acid the degree of inhibition was higher. One of the most potent inhibitors was cysteine. The presence of amino acids does not change the affinity to the substrate of enolase 2-phosphoglycerate. The possible physiological influence of amino acids on the function of enolase is discussed. PMID- 8239905 TI - Kinetic approach and estimation of the parameters of cellular interaction between the immunity system and a tumor. AB - A method is suggested to estimate multi component dynamic systems, which permits, with the help of the computer-calculated kinetic curves, to obtain information about the possible mechanisms of the system component interaction. The method is based on the structural and parametrical identification of mathematical models presented in the form of a system of nonlinear differential equations, using a multi-criterial approach. Using experimental data of studies on growth kinetics and regression of multicellular tumor EMT6 line spheroids in the mouse allogenic system and the immune system cell accumulation in spheroids a mathematical model has been developed of the cellular interaction process in a spheroid. It has been stated that the rate of macrophage and neutrophil accumulation in a spheroid depends on the amount of tumor cells and is determined by the hyperbolic law (as analogous to the Michaelis-Menthen kinetics), while the accumulation of immune lymphocytes in a tumor is determined besides that by the three-cellular cooperation of lymphocytes, macrophages and tumor cells. According to the model, the inhibition of the process of neutrophil and lymphocyte (but not of macrophages) accumulation is realized through the auto-suppression mechanism. The numerical values of the process parameters, which characterise the rates of accumulation, cellular death in a tumor and of local cellular interactions intensity are obtained. PMID- 8239906 TI - Tumor infiltrating lymphocytes in malignant brain tumors. AB - When treated with CD3 moAB (aCD3) (1 microgram/ml) and human natural IL-2 (nIL-2) 1000 U/ml, during the exponential phase of growing: 200 U/ml tumor infiltrating lymphocytes (TIL's) from brain tumors could be expanded. Due to the low percentage of lymphocytes in malignant brain tumor tissue (1%) it was not possible to separate the tumor bearing lymphocytes from the tumor cells by a Ficoll Hypaque gradient separation. This resulted in a co-culture of both lymphocytes and tumor cells in which a serum free medium was used. To generate TIL's at all, it was necessary to supplement this culture medium with 20% of the supernatant of cultures of lymphokine--activated killer cells. In 9 out of 12 cases we obtained total growth factors in the range between 10(4) and 10(14) (average 3.6 x 10(7)). These growth were reached after 6 to 13 weeks including up to 3 weeks of an initial lag period during which no cell growth was observed. We noted a decrease in CD8+ cells, whereby the predominant cell type at the end of the cultures was CD3+ and CD4+. The levels of their cytotoxic activities against K-562 and Raji cells were low in contrast to those of CD3 and nIL-2 stimulated cells which were obtained from carcinomas. PMID- 8239907 TI - Immunoregulation of antitumor response; differential secretion of arachidonic acid metabolites by macrophages during stimulation in vitro with BCG and Corynebacterium parvum. AB - The level of arachidonic acid (AA) metabolites in the supernatants of cultured peritoneal exudate cells (PEC) were studied under various conditions using BCG and Corynebacterium parvum as stimulators. The metabolite levels were analyzed by thin layer chromatography (TLC). The degree of macrophage cytotoxic/cytostatic activity was dependent on the dose and character of stimulators used and the source of macrophages. The application of microcytotoxicity assay for the evaluation of tumor cell lysis (lung sarcoma SaL-1) in vitro revealed that peritoneal macrophages from healthy and tumor bearing BALB/c mice may affect the degree of antitumor response. In the supernatants of cultured PEC from tumor bearing mice AA level increased (by 10-fold) in comparison with PEC from healthy mice. Stimulation with BCG induced over a double level of AA in PEC isolated from tumor bearing mice nonstimulated or stimulated with C. parvum. A lower level of prostaglandins (PGs) was found in the supernatants of cultured PEC isolated from healthy mice (stimulated and non-stimulated), but the highest level of PGs was observed in the supernatants of cultured PEC isolated from tumor bearing mice stimulated with BCG. The unique metabolite of AA was found only in the supernatants from nonstimulated PEC from tumor bearing mice. PEC from tumor bearing mice produced metabolites of AA which were not detected in control group. These results suggest that macrophages also play a regulatory role by secretion of AA. This process can be modified by bacterial antigens. PMID- 8239908 TI - Evaluation of immunophenotype of lymphoid cells isolated from malignant pleural effusions. AB - Lymphoid cells, isolated from malignant pleural effusions and collected from patients bearing primary lung carcinoma, were examined by means of indirect immunofluorescence and a panel of monoclonal antibodies vs several CD antigens. The percentages of CD4+ T lymphocytes were found to be significantly depressed in malignant effusions as compared to inflammatory ones. In relation to histological type of cancer it was especially evident in squamous cell and anaplastic carcinomas (small and large cell), in comparison to adenocarcinomas. Expression of T cell antigen receptor tau/delta (TCR-1) on T lymphocytes, demonstrated by BB3 MoAb (vs V delta 2), was significantly higher in malignant effusions as compared with non-malignant ones. This was not the case when A13 MoAb (equivalent of TCS 1) was used (vs V delta 1). Percentage values of NK cells, monocyte/granulocyte series activated cells and B lymphocytes did not differ significantly in malignant and non-malignant effusions. It is concluded that these are T lymphocyte subpopulations which are apparently distinct in both effusion groups examined. PMID- 8239909 TI - Assessment of immunophenotype of potentially cytotoxic tumor infiltrating cells in laryngeal carcinoma. AB - Among host lymphoid cells engaged in anti-tumor defence, tumor infiltrating cells (TIC) are apparently the most suitable for this purpose, due to feasibility of direct contact with tumor cells. The aim of the present study was to evaluate distribution of TIC which express phenotype of cytotoxic cells, in tissues of laryngeal carcinoma. Cryostat sections of surgical tumor samples were subjected to sensitive APAAP immunohistochemistry, following reaction with one of the panel of monoclonal antibodies (MoAbs) vs. several CD antigens and assessed semiquantitatively. It has been found that the content and distribution of potentially cytotoxic cells are quite heterogenous and vary from case to case in examined cancer. CD8+ cells and those bearing NK cell phenotype were the most frequently encountered, mainly within tumor mass. The cells belonging to NK cell subsets, detected by GL183 and EB6 MoAbs could be demonstrated in tumor proximity. TCR-1+ (tau/delta) T lymphocytes were quite a few in part cases. On the other hand, only a scarce number of cells among TIC expressed interleukin-2 receptor. It is concluded that in the vicinity of laryngeal cancer there are fairly large numbers of potentially cytotoxic cells, but at low or nil state of activation. PMID- 8239910 TI - Production of tumor necrosis factor alpha by resident peritoneal cells of mice. AB - Freshly isolated resident peritoneal cells (RPC) of three mouse strains: BALB/c, NZB and C3H, release spontaneously small amounts of tumor necrosis factor (TNF) when cultured in vitro. RPC of BALB/c mice produced higher TNF levels than the cells isolated from two other strains (BALB/c > NZB > C3H). Lipopolysaccharide (LPS) potentiated the TNF production. Kinetics of the TNF production depended on temperature of incubation of the cells. At 37 degrees C TNF was produced earlier (with a peak at 7 h) than at 26 degrees C (a peak at 20 h). Adherent cell fraction of RPC produced TNF at 26 degrees C as well as at 37 degrees C. Nonadherent cells released TNF only at 37 degrees C. The neutralization assay with media containing TNF and polyclonal rabbit anti-mouse TNF-alpha serum revealed that in every case, independently on the incubation temperature and cell fraction origin, only TNF-alpha was detected. Antibodies against human monoclonal or polyclonal TNF-alpha did not neutralize mouse TNF. PMID- 8239911 TI - Spontaneous and lipopolysaccharide-induced tumor necrosis factor alpha and interferon beta production by resident peritoneal cells of mice: effect of sex, age and strain. AB - BALB/c and NZB mice differ in incidence of autoimmunological disorders. We have studied the dependence of sex and age of these mouse strains on their capacity to produce interferon beta (IFN-beta) and tumor necrosis factor alpha (TNF-alpha). Short term cultures of the mouse resident peritoneal cells (RPC) were used. Three to six week-old female and male BALB/c mice, in contrast to adult mice, did not produce spontaneous IFN-beta (< 2 units/ml) and only low level of TNF-alpha (2-4 units/ml). The levels of lipopolysaccharide (LPS)--induced IFN-beta and TNF-alpha increased progressively with age of BALB/c mice. Female BALB/c mice however, were found to produce approximately two-fold higher levels of IFN-beta and TNF-alpha than male BALB/c mice. Sex dependent differences in IFN-beta production were much more expressed when NZB mice were used in the experiments. RPC of young (3-5 week old) female NZB mice produced relatively high levels of IFN-beta. The spontaneous IFN-beta production by RPC of older female mice (6-8 week-old) declined. In contrast, RPC of young NZB male mice did not produce spontaneous IFN-beta, while RPC of adult male mice were able to release some amounts of IFN-beta. The levels of spontaneous and LPS induced TNF-alpha and LPS induced IFN-beta were apparently not so correlated with sex and age of NZB mice as spontaneous IFN-beta. PMID- 8239912 TI - Production of cytokines by mouse peritoneal cells treated with Tolpa Torf Preparation (TTP): dependence on age and strain of mice. AB - Production of two cytokines: Interferon beta (IFN-beta) and tumor necrosis factor alpha (TNF-alpha) by freshly isolated resident mouse peritoneal cells (RPC) treated with immunostimulating drug Tolpa* Torf Preparation (TTP) was investigated. Nontoxic concentrations of TTP (10 and 100 micrograms/ml) potentiated the cytokine production by RPC of BALB/c mice. The observed effects were dose-dependent. The optimal cytokine-inducing concentration of TTP was 100 micrograms/ml. The effect of TTP depended on the age of mice. The high potentiation (16-fold) of IFN-beta and TNF-alpha production by TTP was observed when 5 and 8 week-old mice were used. However, in RPC isolated from one year old mice, only two (fold) potentiation of IFN and TNF was observed. The activities of the same commercial preparations of TTP were compared in the RPC cultures isolated from mice of the three different strains: BALB/c, C3H/HeJ and NZB. The significant stimulation (16-32-fold) of TNF and IFN was observed in the cells isolated from BALB/c mice. In contrast, only slight stimulation of IFN and TNF (1 3-fold) was observed in cells of NZB mice. Peritoneal cells of C3H mice did not respond to stimulation with TTP. PMID- 8239913 TI - A randomised, double-blind study on the efficacy of Tolpa Torf Preparation (TTP) in the treatment of recurrent respiratory tract infections. AB - TTP is a new immunomodulating drug of natural origin, registered in Poland for human use. In the randomised, double blind study to assess the therapeutic efficacy of the Tolpa Torf Preparation (TTP) in the recurrent respiratory tract infectious participated 39 young patients (age 16-22). TTP was administered orally, 5 mg daily for three weeks. During 3 months follow-up period favourable results of treatment were obtained in 14 of 20 TTP treated patients and in 8 of 19 of the placebo patients. The therapeutic effects were seen even after the 6 months follow-up period. The phagocytic activity of granulocytes was significantly stimulated in the TTP--treated patients but not in the placebo treated patients. The results suggest that TTP is effective drug in the treatment of recurrent respiratory tract infections with undefined infectious etiology. In the period of the treatment as well as during 6 months observation no side effects were noticed. PMID- 8239914 TI - A comparison of efficacy of Tolpa Torf Preparation (TTP) in the treatment of cervicitis with or without surgery. AB - Tolpa Torf Preparation (TTP) is an immunomodulating drug produced by Torf Corporation, Wroclaw and registered for human use in Poland. TTP enhances the process of tissue regeneration. Authors evaluate TTP effectiveness in the treatment of inflammatory states of the cervix, especially cervical erosions and the influence of this preparation of the macroscopic, cytological and bacteriological state of the cervix. TTP was used in 31 patients with the diagnosis of cervical erosion. All patients treated as yet were classified into 3 groups, depending on the treatment of cervical erosion used previously. TTP was administered orally in the dose of 5 mg (in 10 ml of water) daily during 10 days and locally in the form of tampons soaked with 1% TTP solution in the volume of 5 ml also during 10 days. TTP administered this way has beneficial therapeutic effects on the healing of cervical erosion accelerating the process of epithelialization and bringing normalization of the cytological picture. Especially beneficial in the treatment of cervical erosion is combined use of TTP and electrocoagulation or curettage--the healing time can be shortened by half. PMID- 8239915 TI - Quantitation of human haptoglobin by ELISA system based on streptococcal haptoglobin receptors. AB - Streptococcus pyogenes cells with binding properties for human haptoglobin were used for quantitative determination of the acute phase protein, haptoglobin in various biological fluids. The S. pyogenes cells with protein surface antigen T4 served as solid phase in a microtitre ELISA system. After binding to the bacteria the amount of haptoglobin could be quantified by polyclonal or monoclonal antibodies. The constructed ELISA proved to be sensitive and correlated well with a conventional peroxidase method and with an immunoassay based on hemoglobin binding to haptoglobin. PMID- 8239916 TI - Human T cell adhesion to endothelium stimulated by membrane components extracted from strains of Bacteroides vulgatus (member of B. fragilis group). AB - Bacteroides fragilis is reported to be less virulent than other Gram negative rods, existing in normal gut flora (e.g. Enterobacteriaceae). Lymphocytic infiltration, a typical finding in chronic infection, depends on cell extravasation and trafficking towards a site of antigen, a process that is regulated by endothelial expression of adhesion molecules. We have studied the ability of lipopolysaccharides and capsular polysaccharides of B. vulgatus group to induce human T cell adhesion to cultured endothelial cells. Both components of the bacterium caused markedly lesser T cell adhesion than LPS extracted from E. coli. Our data suggest that lower virulence of members of B. vulgatus group might be also dependent on its decreased ability to induce lymphocyte attachment to endothelium and subsequent tissue infiltration. PMID- 8239917 TI - Low dose heparin: efficacious treatment for chronic renal allograft rejection. AB - Recent data indicate that aside from its anti-coagulant action heparin has potent immunobiological activities inhibiting cell trafficking to a site of antigen. In addition, heparin inhibits smooth muscle cell proliferation and decreases the synthesis of the extracellular matrix proteins. As those phenomena are thought to play a major role in the pathogenesis of chronic allograft rejection, we performed a trial evaluating the efficacy of low dose heparin (5 U/kg/day) in 23 recipients of renal allografts with biopsy-proven rejection. In 16 patients (70%) a tendency for improvement was seen which in 57% was statistically significant. Our data suggest that low dose non-anticoagulant heparin may be an efficacious means for treatment of chronic rejection. PMID- 8239918 TI - Vratizolin in treatment of mouth and ear herpetic infections: comparison with conventional therapy. AB - Vratizolin is a new non-steroidal anti-inflammatory drug registered for use on humans in Poland. Published and unpublished data on Vratizolin showed that it has anti-inflammatory, antiviral, antibacterial, antimycotic, analgesic and immunomodulating activities. The purpose of these randomized, parallel-group studies was to compare Vratizolin with other standard drugs, used for the treatment of mouth and ear infections. The study involved 193 patients with recurrent Herpes simplex, Herpes zoster oticus, Stomatitis herpetica and infections of the external ear canal. Vratizolin was used topically, as 3% hydrophilic cream or ointment, four times daily. Standard treatment included zinc ointment, Aphtin (boric acid plus glycerin), Oxycort and Dicortinef. In almost all of the treated patients the efficacy of Vratizolin treatment was superior to the drugs mentioned above. It was assessed by measuring disappearance of both objective (edema, erythema, crusting) and subjective symptoms (pain, burning and itching). PMID- 8239919 TI - Hematological effects of the new immunosuppressive drug 15-deoxyspergualin. AB - Since systematic hematological studies on blood and bone marrow changes after treatment with 15-Deoxyspergualin (DOS) are lacking, a quantitative assessment was performed fourteen or twenty eight days after intraperitoneal application of DOS to rats. Further observations done 7 and 14 days after discontinuation of DOS administration allowed analysis of bone marrow regeneration. DOS induced lymphocytopenia, granulocytopenia and anemia with a decrease of bone marrow cellularity due to suppression of cell maturation. The effect was dose-dependent and bone marrow as well as blood changes were observed in animals treated with doses from 0.5 to 10.0 mg/kg DOS. Within 14 days after termination of the treatment, rapid recovery with normalization of all hematological parameters was observed. In the light of our data, these hematological side effects may not be a major disadvantage, if DOS is used in doses below 2.5 mg/kg, and for a course of therapy which is limited to 7 to 14 days. PMID- 8239920 TI - Induction of tumor necrosis factor and interleukin 6 by outer membrane proteins of Shigella in spleen cells and macrophages of mice. AB - The ability of outer membrane proteins (OMP) of Shigella flexneri to induce tumor necrosis factor (TNF) and interleukin 6 (IL-6) production by spleen cells and macrophages of mice was investigated. Treatment of spleen cells with OMP resulted in the release of only traces of TNF activity. In contrast, macrophages treated with OMP produced moderate levels of TNF. OMP was also found to be an inducer of IL-6. Both spleen cells and macrophages, treated with OMP, were found to produce substantial levels of this cytokine. The effect of OMP on the release of TNF and IL-6 was dose and time dependent, maximal production being reached at 10 micrograms of OMP after 20 h. The ability of OMP to induce production of these cytokines may explain part of the previously described immunomodulatory effects of this preparation on the immune system. PMID- 8239921 TI - Comparison of the functional activities of two different preparations of human recombinant tumor necrosis factor alpha. AB - The role of tumor necrosis factor (TNF-alpha) in physiological and pathological reactions has resulted in a progressive increase of expensive TNF-alpha consumption for laboratory and clinical purposes. Following this trend, the first chemical synthesis of the gene for rHuTNF-alpha gene in Poland and its subsequent successful expression in E. coli was recently reported. In the present paper, we verify the in vitro biological activities of this TNF-alpha preparation (CMMS/TNF alpha) in comparison with a commercially available preparation of TNF-alpha. We demonstrate that our TNF-alpha possesses strong cytotoxic activity against WEHI 164 (clone 13) cells, binds the p55 and p75 TNF receptors on cell lines, induces intercellular adhesion molecule-1 (ICAM-1) expression, and interleukin-6 (IL-6) and granulocyte-macrophage colony-stimulating factor (GM-CSF) release from human umbilical vein endothelial cells (HUVEC). We demonstrate its usefulness for further investigations as an effective reagent for in vitro assays. PMID- 8239922 TI - Detection, by in situ hybridization using sulphonated cDNA probe, the specific mRNA for HLA-DR alpha induced in monocyte cell lines by recombinant interferon gamma. AB - We adopted the nonradioactive method used for blot hybridization for the detection of inducible mRNA for HLA-DR alpha by the in situ hybridization. Unstimulated and interferon gamma stimulated MonoMac6 and U937 human monocytic cell lines were used as target cells. Sulphonation of plasmid pBR322 with HLA-DR alpha cDNA insert (2 x 700 bp, in Pstl restriction site) was performed according to the manufacturer's procedure (SulfoProbe Kit, Sigma). The hybridization signals were detected with mouse monoclonal, anti-sulphonated DNA antibody, followed by immunovisualization with anti-mouse IgG-alkaline phosphatase conjugates. Unstimulated MonoMac6 and U937 cells showed few granular reaction products only in small percentage of cells (1-5%), while in IFN gamma stimulated cells the fine granular immunoenzymatic reaction was observed in the cytoplasm of majority of cells (> 80%). This method seems to be easy and rapid to perform, making it applicable for routine diagnostic purposes in tissue sections and biopsies. PMID- 8239923 TI - Flawed creatine kinase utilization study. PMID- 8239924 TI - Transbronchial lung biopsy after lung transplantation. PMID- 8239925 TI - Clinical usefulness of DNA ploidy and cell cycle studies. PMID- 8239926 TI - Bedside monitoring of blood glucose in hospitals. Speed vs precision and accuracy. PMID- 8239927 TI - Bedside glucose monitoring. A College of American Pathologists Q-Probes study of the program characteristics and performance in 605 institutions. AB - In 1991, the College of American Pathologists' Q-Probes quality improvement program studied precision, accuracy, and program characteristics of bedside glucose monitoring (BGM) in 605 institutions. Precision measurements made in 569 institutions were based on 15,950 quality control results. Precision, expressed as a percentage coefficient of variation, was less than 10 in almost 90% of the institutions. For accuracy, 4517 BGM results from 181 institutions were compared with clinical laboratory glucose results. Approximately 58% of BGM results were within +/- 10% and 75% of BGM results were within +/- 15% of the corresponding clinical laboratory results. Program characteristics associated with better performance were (1) laboratorian program responsibility, laboratorian BGM test performance, and laboratorian involvement in training; (2) a policy requiring repeated scheduled operator training and/or performance review; (3) use of an internal comparison program; and (4) participation in an external proficiency testing program. We conclude that BGM frequently does not meet quality goals and provide recommendations on how BGM programs can be improved. PMID- 8239928 TI - DNA-based identification and epidemiologic typing of bacterial pathogens. AB - Genotypic, or DNA-based, methods have become increasingly applicable for infectious disease diagnosis and epidemiologic analysis. The ability to assess the pathogen's genotype directly bypasses requirements for cultivation and may diminish diagnostic delays with fastidious organisms. Genotypic typing methods have enhanced epidemiologic studies by providing techniques with greater discriminatory ability and smaller proportions of nontypeable samples. Hence, these methods enable rigorous studies to be performed regarding the nature of disease outbreaks. Phenotypic, or conventional, methods will remain important in diagnostic microbiology, especially with organisms that are relatively easy to culture. However, genotypic methods may become increasingly prominent in clinical microbiology laboratories, particularly with respect to diagnosis of fastidious pathogens, because of their relative speed, versatility, and lack of cultivation requirements. PMID- 8239929 TI - Polymerase chain reaction-based detection of B-cell monoclonality in cytologic specimens. AB - Thirty-seven cytologic cell blocks were evaluated for B-cell monoclonality by polymerase chain reaction (PCR), 16 of them cytologically positive for lymphoma, and 21 suspicious for lymphoma but morphologically nondiagnostic. Of 37 specimens, 13 (35%) showed B-cell monoclonality, including six of 16 cytologically positive samples and seven of 21 cytologically suspicious ones. Of these 13 positive samples, seven were positive using crude lysates as substrates, and six additional positive samples were identified only when DNAs were purified and concentrated. Analysis of the DNAs further revealed poor polymerase chain reaction amplifiability and low DNA yield in many samples, indicating that cell block materials are suboptimal for this assay. We concluded that B-cell monoclonality can be detected in ethanol-fixed cytologic samples, and usage of unembedded material will likely improve the sensitivity. In specimens cytologically suspicious for lymphoma, polymerase chain reaction-based identification of monoclonal B-cell population supports the diagnosis of B-cell lymphoma and is a potentially useful test in solving this diagnostic dilemma. PMID- 8239930 TI - The impact of new DNA diagnostic technology on the management of cancer patients. Survey of diagnostic techniques. AB - Revolutionary advances in technology have enhanced our understanding of the genetic changes that occur in cancer cells. This article summarizes some of the basic features of these techniques and describes their application to the identification of specific types of genetic alterations in cells. The emphasis is on their use in obtaining information that is of diagnostic and prognostic importance. The techniques fall into two broad categories; the first is the direct analysis of the chromosome pattern in metaphase cells or the indirect assessment of chromosome abnormalities in interphase nuclei. The second general category involves the isolation of DNA, RNA, or protein from the tumor cells and the analysis of these components for abnormalities related to the presence, absence, or amplification of a specific gene or its products or other alterations, eg, those due to chromosome translocations. The techniques described in this article have broad applicability to medicine in general and some familiarity with these techniques is critical for the practice of modern medicine. PMID- 8239931 TI - Unbiased analysis of the frequency of beta-thalassemia point mutations in a population of African-American newborns. AB - OBJECTIVE: To determine frequency of specific beta-thalassemia alleles in the African-American population prospectively, using newborn screening specimens, and to evaluate the need for including these alleles in screening follow-up programs. DESIGN: Allele-specific oligonucleotide tests were developed and used to analyze African-American newborn screening specimens for beta-thalassemia point mutations to determine their frequency. Direct sequencing of amplified DNA from the dried blood specimens was used to confirm the presence of point mutations. POPULATION: African-American newborns in Texas. RESULTS: Allele-specific oligonucleotides identified five newborn specimens carrying beta-thalassemia point mutations among 471 specimens from African-American neonates. Direct sequencing of DNA from the dried blood specimens confirmed that these individuals had a normal and a mutant allele. Five newborn screening specimens in which the results of screening and DNA tests were in disagreement (four with FS by screening and AS by DNA, and one with FC by screening and AC by DNA) were analyzed for these beta-thalassemia point mutations and in each case were found to be S/beta-thalassemia or C/beta thalassemia compound heterozygotes, respectively. CONCLUSIONS: Allele-specific oligonucleotides accurately identified newborn specimens carrying beta thalassemia point mutations. Direct sequencing from dried blood specimens confirmed these results. The A(-29)G allele frequency was 0.003, and the C(-88)T frequency was 0.002. These alleles also were observed among positive samples in a neonatal hemoglobinopathy screening program. Therefore, any newborn screening program with a molecular genetic follow-up component must include testing for these beta-thalassemia alleles to assure timely and appropriate management for affected infants. PMID- 8239932 TI - Detection of Epstein-Barr virus in human tissues by molecular genetic techniques. AB - In the past few years, there has been an explosion of new data on the association of Epstein-Barr virus (EBV) with human disease. Many of these discoveries have come as a direct result of the application of DNA technology. The nucleic acid hybridization techniques most commonly used to detect EBV in human tissues include Southern blot analysis, in situ hybridization to viral DNA or RNA, and polymerase chain reaction. An advantage of Southern blotting is the ability to distinguish latent from infectious EBV and to determine the clonality of infected tumors with respect to the structure of the viral terminal repeat sequences. In situ hybridization has the advantage of precise localization of the virus in infected tissues or tumors. Polymerase chain reaction is exquisitely sensitive in detecting viral DNA, perhaps too sensitive for disease-specific purposes given the ubiquitous nature of EBV. Each of these molecular genetic methods of EBV analysis is currently used in research laboratories, while some methods have found their way into routine diagnostic pathology because they are faster, more sensitive, or more informative than previous assays. PMID- 8239933 TI - Molecular pathology of the fragile X syndrome. AB - Fragile X syndrome is the most common form of familial mental retardation (one in 1250 males and one in 2500 females, characterized by prominent dysmorphic features, macro-orchidism, and varying degrees of mental retardation. Diagnosis of this syndrome has relied on cytogenetic demonstration of the fragile site at position Xq27.3. A gene associated with the fragile X syndrome, FMR-1, has been isolated and mapped to the region of the X chromosome that corresponds to the region of the fragile site. Expansion of a trinucleotide repeat, CGG, and abnormal methylation of a CpG island account for the majority of mutations identified in FMR-1. These molecular characteristics have greatly enhanced the identification of affected individuals and carriers of the premutation who were not detected cytogenetically. PMID- 8239934 TI - DNA ploidy of ameloblastoma and ameloblastic carcinoma of the jaws. Analysis by image and flow cytometry. AB - Ameloblastomas, the second most common odontogenic tumor, behave as benign locally aggressive tumors. Ameloblastic carcinomas, on the other hand, show histologic features of malignancy and may metastasize. By image cytometric analysis of Feulgen-stained sections from decalcified, formalin-fixed, paraffin embedded tumors, we compared nuclear DNA content of 17 primary and five recurrent ameloblastomas and five ameloblastic carcinomas. Of the primary ameloblastomas, 14 (82%) were diploid; three (60%) of the five recurrent ameloblastomas were diploid. No significant difference in ploidy between primary and recurrent ameloblastomas or among plexiform, follicular, and acanthomatous types of ameloblastoma was demonstrated. Of the five ameloblastic carcinomas, four (80%) were aneuploid; ploidy was not significantly correlated with the incidence of metastasis. Flow cytometry was performed on three carcinomas; 100% concordance between image and flow cytometric data was seen. Aneuploidy is significantly more common in ameloblastic carcinomas than in ameloblastoma and is a strong predictor for malignant potential. PMID- 8239935 TI - Concordance of DNA content between prostatic intraepithelial neoplasia and concomitant invasive carcinoma. Evidence that prostatic intraepithelial neoplasia is a precursor of invasive prostatic carcinoma. AB - It has been postulated that prostatic intraepithelial neoplasia (PIN) is a precursor lesion to invasive carcinoma in the prostate, yet there has been little direct evidence for this relationship. Therefore, we studied the DNA content of the epithelial cells in PIN lesions with accompanying invasive carcinoma in seven cases. DNA content of nuclei in Feulgen-stained tissue sections was performed using the CAS200 System (Cell Analysis Systems Inc, Elmhurst, Ill), with normal and hyperplastic glands serving as controls for diploid DNA content. In four cases, the cells of both lesions (PIN and invasive carcinoma) contained only diploid DNA; in one case, an additional aneuploid tumor stem line was present only in the invasive component; and in two cases, both PIN and carcinoma cells contained aneuploid cells having similar DNA content. The concordance of DNA content between PIN and invasive carcinoma in these cases provides evidence that PIN is a precursor lesion for invasive carcinoma. PMID- 8239936 TI - A method for detecting variability arising from errors in sample processing of paraffin-embedded tissue for DNA content analysis. AB - We present a method for controlling variability that may arise from inconsistencies in sample preparation for DNA content analysis of paraffin embedded tissue. Human tonsil tissue obtained from routine surgical specimens was embedded in paraffin according to standard protocols. Fifty-micrometer sections were cut from the block and analyzed each day for 20 days to establish control ranges. One tonsil tissue section was processed in parallel with each run of clinical specimens. In this context, a run was defined as the simultaneous processing of 50-microns tissue sections for extraction of cell nuclei (dewaxing and rehydrating). If the tonsil G0/G1 peak coefficient of variation (CV) exceeded 2 SDs of the established mean, and optimum instrument performance and staining were verified, all samples prepared with the tonsil control were reprocessed. Instrument performance and staining were assessed by using the appropriate external controls. By using this rejection rule (12s), the frequency of sample reprocessing in our laboratory was approximately 6%. When the run was repeated and the tonsil control CV was within acceptable range, the G0/G1 peak CV of the corresponding clinical specimens improved 25% of the time. Because most investigators are willing to accept higher CVs for paraffin-embedded tissue than for fresh tissue, it is desirable to have a control to detect decreased peak resolution, resulting from errors in sample processing. PMID- 8239937 TI - Flow cytometric DNA analysis for the determination of malignant potential in adrenal and extra-adrenal pheochromocytomas or paragangliomas. AB - DNA ploidy studies were performed by flow cytometry on extracted nuclei from 53 adrenal, 13 carotid body, 14 retroperitoneal, two intrathoracic, two urinary bladder, and one cauda equina pheochromocytomas (paragangliomas). A specific technique was used for preparation of paraffin-embedded tissue into single associated nuclei, and another specific method was used for propidium iodide staining. Twenty normal adrenal glands were also analyzed as controls. Six tumors in the adrenal medulla yielded uninterpretable histograms (coefficient of variation > 7%) because of excessive cell debris or other technical failure. All control adrenal glands were diploid. Seventy-two tumors were clinically benign and seven were malignant as evidenced by regional or distant metastases and/or extensive local invasion. Fifty-six (71%) of 79 tumors disclosed a normal DNA diploidy pattern with a benign clinical course. Abnormal DNA histograms including tetraploidy and aneuploidy patterns were observed in the remaining 23 cases, including the seven malignant tumors. Significantly more malignant tumors occurred in the DNA tetraploid group and the DNA aneuploid group than in the normal DNA group; considered together, the two groups with abnormal DNA histograms differed significantly from the normal DNA group. On the basis of these results, nondiploid tumors are considered to be more prone to aggressive behavior than diploid tumors and, therefore, should be carefully monitored. PMID- 8239938 TI - Quality of laboratory performance in testing for human immunodeficiency virus type 1 antibody. Identification of variables associated with laboratory performance. AB - To identify factors that may affect the quality of laboratory performance of human immunodeficiency virus type 1 (HIV-1) antibody testing, the Centers for Disease Control and Prevention Model Performance Evaluation Program surveyed laboratories in 1989 that performed enzyme immunoassay (EIA) and Western blot tests for HIV-1 antibody. Panels of 10 HIV-1-antibody-positive and antibody negative plasma samples, some of which were duplicates, were mailed to program participating laboratories. Laboratories were also mailed survey questionnaires to ascertain their laboratory characteristics and testing practices. Using 1988 data, researchers previously found that the overall analytic performance of laboratories performing HIV-1 antibody testing was independently associated with the following: (1) requiring a minimum degree of testing personnel; (2) having written criteria for identifying unsatisfactory specimens; (3) requiring in-house training for testing personnel; (4) having tested more than 10,000 specimens; (5) being identified as an "other" laboratory type; (6) having more than 24 months of testing experience; (7) laboratory uses specific (Abbott) materials for EIA; and (8) testing specimens collected by family-planning clinics. To verify these findings, we performed multivariate analysis on 1989 performance data. For the 1989 EIA analytic sensitivity, significant positive (P < or = .05) associations were detected with having written criteria for identifying unsatisfactory specimens and with having tested more than 10,000 specimens. For the 1989 overall EIA analytic performance, a significant negative (P < or = .05) association was found with using specific (Abbott) EIA materials, and a significant positive (P < or = .05) association was found with having tested more than 10,000 specimens. For Western blot results, the only significant (P < or = .05) associations were for both analytic sensitivity and overall analytic performance and having tested more than 10,000 specimens. PMID- 8239939 TI - Adenocarcinoma of renal pelvis. AB - Adenocarcinoma accounts for a small percentage of neoplasms arising within the renal pelvis. We describe a mucinous adenocarcinoma of the renal pelvis that occurred in a 57-year-old woman. Investigation of the recent literature reveals an additional 12 cases of adenocarcinoma of the renal pelvis reported since 1980. These 13 cases are summarized in detail, for a total of 59 cases of adenocarcinoma of the renal pelvis documented in the English-language literature. These tumors can be subdivided into three major histologic types: tubulovillous, mucinous, and papillary non-intestinal. The tubulovillous and mucinous tumors are morphologically similar to intestinal tumors accounting for 71.5% and 21.5% of cases, respectively. They are believed to arise in foci of intestinal metaplasia. Only three cases (7%) were of the nonintestinal, nonmucinous, papillary subtype. These rare tumors are notable for their morphologic similarity to Bellini or collecting duct carcinoma, but a specific morphologic precursor has not been identified. Of the three subtypes, tumors of tubulovillous morphology confer the worst prognosis with 70% of patients dying within 5 years. Thirty-three percent of mucinous tumors and none of the papillary nonintestinal tumors were fatal. PMID- 8239940 TI - Juxtaglomerular cell tumor. AB - We report the cytopathologic, immunocytochemical, histopathologic, and electron microscopic findings of a juxtaglomerular cell tumor that developed in a hypertensive 25-year-old Japanese woman. The symptoms of her disease disappeared after the tumor was resected. To our knowledge, this is the first case report of this rare tumor, which was diagnosed after imprint cytology of the resected tumor. The clinical diagnosis was made before surgery based on the high level of plasma renin activity. Imprint cytology showed knob-like formations of neoplastic cells that exhibited a positive reaction to human renin antigen. Histologic and electron microscopic examination revealed the typical features of juxtaglomerular cell tumor as well as the tubular components that are believed to originate in the distal urinary tubules. PMID- 8239941 TI - Origins of adenocarcinomas presenting as intracranial metastases. An ultrastructural study. AB - Adenocarcinomas of unknown origin are a common neurosurgical problem. In many such cases, the primary tumor remains clinically obscure. A series of 28 adenocarcinomas of unknown origin, presenting as intracranial metastases and examined with electron microscopy, was reviewed blindly in a "retrospective prospective" fashion. Distinguishing ultrastructural features allowed prediction of site of origin in 26 cases (93%). Clinical and postmortem follow-up allowed the identification of likely primary tumor sites in 13 cases, with confirmation of the ultrastructural diagnosis in all 13 cases. The great majority of adenocarcinomas of unknown origin were found to originate in the lung (22/26 [85%]), while a few originated in the colon (2/26 [8%]) or kidney (2/26 [8%]). The predominance of pulmonary tumors among intracranial metastatic adenocarcinomas of unknown origin is much greater than the reported 45% to 55% predominance of pulmonary tumors among all intracranial metastatic carcinomas of unknown origin. Electron microscopy is a useful technique to either confirm a bronchogenic origin or to suggest an alternative primary site for these tumors. PMID- 8239942 TI - Aortic dissection in cardiac allograft recipients. A report of two cases. AB - Aortic dissection in cardiac allograft recipients has not been reported previously, to our knowledge. We have observed two fatal aortic dissections in heart transplant recipients: one in the immediate postoperative period and one occurring 5 years after transplantation. We discuss the clinical presentations, pathologic findings, possible pathogenetic mechanisms, and risk factors. Because the frequency of aortic dissection after heart transplantation appears to be relatively low (< 1%), such disease may be clinically unsuspected. Given the uniformly poor outcome observed in our two patients, we suggest that increased awareness of the possible development of aortic dissection after heart transplantation may result in improved survival of affected patients. PMID- 8239943 TI - Quality control of immunocytologic testing. Prolonged preservation of cell surface antigen reactivity with magnesium chloride-sucrose solution. AB - A sucrose-magnesium chloride-glycerol storage solution (sucrose-magnesium chloride) can preserve antigenicity of cytologic preparations for prolonged periods of time. This storage method was evaluated relative to our immunocytologic technology with the specific aim of exploring its use as a quality control measure. Neuroblastoma and leukemia cell lines, patient bone marrow, and blood specimens were serially tested during a 15-week period to evaluate preservation of immunoreactivity and cell morphological features. Slides that had been air dried and stored at 4 degrees C were compared with those that had been fixed with methanol and paraformaldehyde and stored in sucrose-magnesium chloride at -20 degrees C. The sucrose-magnesium chloride method proved to be superior, and antigen-antibody binding and cell morphology were preserved even after 15 weeks of storage. This method, now in use as a quality control measure in our laboratory, constitutes a practical assurance program for immunocytologic analysis. PMID- 8239944 TI - [Synthesis and CNS-activity of spirocyclic pethidine and prodine analogs]. AB - The bromoacetals 5a and 5b react with n-butyllithium and the piperidone 7 to yield the hydroxyacetals 8b and 8c, respectively. Cyclization of 8b and 8c followed by acid hydrolysis affords the spirocyclic hemiacetals 10b and 10c which are oxidized by PCC to give the spirocyclic prodine analogues 4b and 4c. The corresponding spirocyclic pethidine derivative 2 is prepared by alkylation of the 2-benzopyran-3-one 16 with N-Lost (17). In the mouse writhing test the spiropethidine 2 is not analgesic active up to a dose of 20 mg/kg body weight (bw). In the spirocyclic prodine series the methylated lactone 4c is the most active analgesic with an ED50-value (ED50 = 9.2 mg/kg bw) in the range of the ED50-value of tramadol. PMID- 8239945 TI - Cyclic ureas, III: On the metabolism of the enantiomers of cyclic ureas in rats. AB - Studies are described on the metabolism of the enantiomers of 1, 2, and 3 in rats. The metabolites were identified in urine after cleavage of conjugates and extraction using gas chromatography-mass spectrometry. Qualitative enantioselective differences of the metabolism of 1, 2, and 3 could not be detected. Because oxidation of 1 and 2 to the corresponding barbiturates could not be found and 3 was oxidized to mephenytoin only in minor amounts, this pathway is not a prerequisite for the sedative-hypnotic effects of 1, 2, and 3. PMID- 8239946 TI - Antifungal agents, II: Synthesis and antifungal activities of aryl-1H-pyrrol-2-yl 1H-imidazol-1-yl-methane derivatives with unsaturated chains. AB - The synthesis and antifungal activities of aryl-1H-pyrrol-2-yl-1H-imidazol-1-yl methanes having allyl, crotyl, and acrylate chains linked to the N-pyrrole atom and substituted at phenyl ring by Cl, F, CH3, and NO2 groups are reported. In vitro tests against Candida albicans and Candida spp. showed 2,4-dichlorophenyl-1 allyl-1H-pyrrol-2-yl-1H-imidazol-1-yl-methane to be the most potent derivative with activities comparable to those of ketoconazole and slightly inferior to those of bifonazole and miconazole. Some structure-activity relationships are discussed. PMID- 8239947 TI - Sydnone derivatives: synthesis and antimicrobial activity. AB - Amongst a number of mesoionic compounds studied so far, sydnones have attained importance due not only to their structural features and chemical properties, but also to their biological properties. Sydnone derivatives have been extensively studied for their biological activities, the antimicrobial activity of substituted aryl sydnones has been reported. Aryl sydnones are less toxic and more active than alkyl sydnones. Also chalcone analogues are known to have antimicrobial activity. PMID- 8239948 TI - Cardiac risk factors immediately following spinal injury. AB - Cardiac risk factors were evaluated in 48 persons (39 males, 9 females) with quadriplegia or paraplegia, resident in a specialized spinal injury hospital and seen 0.3 +/- 0.7 years after their spinal injury. The majority of the patients reported being extremely active physically prior to injury. Limited activity in the hospital involved the use of ergometers, pulleys, and weights; however, the majority of subjects expressed a wish for more exercise. Serum lipid profiles showed a relatively normal total cholesterol (mean 4.58 +/- 0.77 mmol/L), a very low high-density lipoprotein-cholesterol (0.91 +/- 0.27 mmol/L), a relatively normal low-density lipoprotein-cholesterol (2.86 +/- 0.68 mmol/L) and high triglycerides (1.89 +/- 0.88 mmol/L). Resting blood pressures were normal. The percentage of smokers (25%) was similar to the percentage of smokers in the general population, but many had quit smoking subsequent to hospitalization. Family histories and diet gave no evidence of increased cardiac risk. We conclude that individuals who sustain a spinal injury do not have a large inherent risk of cardiac events. In persons with paraplegia, cardiac problems develop mainly from the cumulative impact of reduced physical activity and a resulting adverse lipid profile. There may also be a reactive deterioration in other aspects of personal lifestyle, such as cigarette smoking after leaving the hospital. In high-level lesions, factors such as hypertension and a poor stroke volume with compensatory tachycardia may increase cardiac work rate during attempts at ambulation, further predisposing an individual to myocardial ischemia and cardiac arrest. PMID- 8239949 TI - Influences of height and gender on normal nerve conduction studies. AB - Our purpose was to examine the effects of gender on nerve conduction results in healthy subjects. Fifty-four men, mean age 60.2 years and mean height 167cm, were compared with 62 women, mean age 62.2 years and mean height 153cm. We studied median, ulnar, sural, peroneal, and tibial nerves. Analyzing the raw data, women had significantly faster conduction velocities than men for all nerves except median motor (p < 0.05) three of four sensory amplitudes were larger in women and two of four motor amplitudes were larger in men (p < 0.05). After adjustment of the data for height, most statistically significant differences in conduction velocity disappeared, although differences in amplitude persisted. We conclude that most gender differences in nerve conduction velocity can be largely explained by height, whereas amplitude differences persist despite correction for height, temperature, and age. PMID- 8239950 TI - Reliability determination of F mean response latency. AB - Two hundred consecutive ulnar F wave latencies were evaluated in 30 healthy controls (C) and in 9 patients with mild neuropathy (N). The primary study objective was to determine the F wave sample size required to obtain a reliable F mean latency. A secondary objective was to explore the diagnostic utility of three F wave parameters: the minimal F wave latency (Fmin), the average F wave latency (Fmean), and the minimal-maximal F wave latency difference (Frange). For the control group, the Fmean reliability estimates as measured by intraclass correlation coefficients were 0.87, 0.97, and 0.99 with 1, 5, and 10 responses, respectively; for the N group, the corresponding values were 0.73, 0.93, and 0.96. Fmean was abnormal in 89% of the subjects with mild neuropathy whereas Frange and Fmin were abnormal in 67% and 33%, respectively. The study suggests (1) a reliable F mean latency can be obtained by averaging ten F responses; and (2) Fmean and Frange offers promise in increasing the diagnostic utility of the F response in identifying mild neuropathies. PMID- 8239951 TI - Intrasubject variability of selected force-platform parameters in the quantification of postural control. AB - Platform stabilometry is increasingly applied to monitor or re-educate standing balance in clinical rehabilitation. Consequently, insight is needed into the validity, reliability, and sensitivity of different force-platform parameters. This study focuses on the intrasubject variability as the major source of variance (unreliability) in the study of human motor skills. The intrasubject variability of several, commonly applied force-platform parameters was determined across ten repeated tests of quiet two-legged standing in healthy subjects to identify the most consistent and stable parameters in the quantification of postural control. The variability of the root mean square (RMS) amplitude, peak to-peak amplitude, mean frequency, and RMS velocity of the fore-aft and lateral components of the center-of-pressure fluctuations was investigated under varying (visual and cognitive) task conditions. The results indicate that all selected parameters show considerable intrasubject variability irrespective of the task context. Nonetheless, both the RMS amplitude and RMS velocity in either direction of sway do not demonstrate a significant trend across repeated tests. Among the selected parameters, the RMS velocity in the fore-aft direction shows the greatest intrasubject consistency, as well as a high sensitivity to, for example, visual deprivation. These findings support the reliability and validity of this parameter in the clinical quantification of postural control. PMID- 8239952 TI - Posturographic measurement of body sway in survivors of severe closed head injury. AB - Postural imbalance was measured in 39 survivors of severe closed head injury 7 to 66 months after head injury using a posturographic platform. The results were compared with those derived from age-matched healthy control subjects. Severe postural imbalance, particularly in an anteroposterior direction, was found in 16 patients, whereas 9 patients showed moderate imbalance and 14 patients showed normal results in the posturographic investigation. Low initial Glasgow Coma Scale scores and deep parenchymal brain lesions demonstrated by magnetic resonance imaging were shown to be significant indicators of subsequent severe postural imbalance. The duration of posttraumatic amnesia, the localization and size of cortical contusions and subcortical white matter lesions, on the contrary, were not associated with postural imbalance in the long-term outcome. PMID- 8239953 TI - Changes in medical stability upon admission to a rehabilitation unit. AB - The implementation of Medicare's prospective payment system in acute care has coincided with a steady increase in medically unstable admissions to our freestanding rehabilitation facility. We investigated the consequences of these admissions by collecting medical information regarding transfers beginning in 1983. Patients requiring transfer back to the acute setting within 1 day of admission were considered medically unstable and their charts were reviewed. The number of patients requiring transfer back within 1 day increased from 1.5% of all first admissions to 3.1% in 1988 (Mantel - Haenszel chi 2 = 8.03, (df = 1), p < .01), but the increase among Medicare patients alone was not significant. This progressive increase was most pronounced in the cerebrovascular accident and spinal cord injury populations. Beginning in 1988, an intensified preadmission evaluation program was implemented, resulting in a significant decline in unstable patient transfers from hospitals where our consultants were on staff. Physiatric consultations at referral institutions decreased the number of unstable patients at admission. PMID- 8239954 TI - Long psychomotor reaction time in patients with chronic low-back pain: preliminary report. AB - The hypothesis in this study was that slow psychomotor reaction time is related to low-back pain. A total of 73 people with chronic low-back pain (CLBP) were studied. Simple reaction time (SRT) and choice reaction time (CRT) were measured. CRT was also fractionated into decision and movement times. Each four variables measured were analyzed by using analysis variants and covariants with two grouping factors (CLBP and gender) with age and height serving as covariates. The CLBP group had longer SRT, decision time, and total CRT than the control group on average. The results did not conflict with our hypothesis. Slow psychomotor speed of reaction may be one factor in the development of CLBP, or slow reaction in CLBP may have resulted from fear responses, depression, or anxiety. Longitudinal studies are needed in the cause-and-effect evaluation of the relation between speed of reaction and CLBP. PMID- 8239955 TI - Muscle strengthening in a post-polio subject through a high-resistance weight training program. AB - A 48-year-old woman started experiencing increased muscle weakness and fatigue at age 44, 40 years after the onset of acute poliomyelitis. The acute poliomyelitis resulted in severely weak lower limb muscles and as a result, she had used crutches for more than 40 years. Computer-aided force transducer systems were used to determine isometric muscle strength. Both quadriceps and left ankle dorsiflexors were severely weak and could not generate any measurable force. Isometric muscle strength of her right ankle dorsiflexors was 77 Newtons (N) (approximately 1/3 normal); isometric muscle strength of right elbow flexors (171N) and left elbow flexors (160N) were within normal range, although she complained of weakening based on her inability to climb two stairs at a time with crutches as she was used to. She underwent high-resistance weight training of her right ankle dorsiflexors and left elbow flexors for one year. Weight training was three times per week, five sets of ten repetitions per session; total duration of muscle contraction (excluding rest periods) was two and one half minutes per session, 30 minutes per month. Muscle strength (N) was remeasured after four, eight, and 12 months. Muscle strength of right ankle dorsiflexors increased by 61%, whereas that of the left elbow flexors increased by 32% after one year of weight training. She also expressed a subjective feeling of increased muscle strength. High-resistance, short duration muscle strengthening exercise programs should therefore be given a serious consideration in the rehabilitation management of moderately weak muscles of post-polio subjects. PMID- 8239956 TI - Serial manual muscle testing in Duchenne muscular dystrophy. AB - Serial measurements of muscle strength of 34 muscle groups were obtained for up to a seven-year period using manual muscle testing (MMT) in a group of 63 boys (mean age = 11.5 years) with Duchenne muscular dystrophy (DMD) not using lower extremity bracing or corrective surgery for ambulation, in order to define the natural course of strength loss and its relation to anthropometric variables and ambulatory status. When correlating the first MMT assessment of each patient with age, strength was found to decrease linearly (-0.26 +/- 0.30 MMT units/year) with time until approximately age 14 when a marked decrease in progression was noted ( 0.06 +/- 0.03 MMT units/year). Analyzing individual patients longitudinally yielded a similar rate of strength loss. The average muscle score was 3.0 MMT units at the point of full-time wheelchair use. The marked decline in MMT deterioration around age 14 may be an important consideration when assessing the efficacy of therapeutic interventions in adolescent boys with DMD. PMID- 8239957 TI - Pressure ulcers in community-resident persons with spinal cord injury: prevalence and risk factors. AB - The prevalence and correlates of pressure ulcers in terms of their number, severity, and anatomical location were studied in a community-based sample of 100 men and 40 women with spinal cord injury. Thirty-three percent (n = 46) presented with one or more ulcers of at least one stage I severity when visually examined. Twenty-one individuals had more than one ulcer, the maximum number of ulcers being seven. Of 87 ulcers for which severity ratings were available, 30 (34.5%) were stage I, 33 (37.9%) were stage II, and 24 (27.6%) were either stage III or IV. Individuals with an ulcer exhibited more paralysis and were more dependent on others in activities of daily living. A greater proportion of blacks had more severe ulcers (stages III and IV) than their white counterparts. Persons with more severe ulcers incurred their injury later in life, and had significantly lower mean scores on the Occupation and Mobility dimensions of the Craig Handicap Assessment and Reporting Technique. The findings suggest that factors governing initial development of a pressure ulcer differ in part from those responsible for an ulcer progressing in severity. PMID- 8239958 TI - Physical training effects in myasthenia gravis. AB - Eleven patients with mild or moderate myasthenia gravis (MG) were subjected to a strength training program of 27 to 30 sessions during ten weeks. Voluntary maximal muscle force and the degree of fatigue during repetitive maximal isometric muscle contractions were measured in three muscle groups. The subjects did not experience any subjective discomfort or any adverse effect on their MG due to the training. A 23% increase of the maximal voluntary muscle force in knee extension in the trained side was found, compared to 4% in the untrained side (p < 0.05). Only small changes were noted regarding maximal muscle force in elbow flexion and extension or in any of the muscle groups in the fatigue test. Physical training can be carried out safely in mild MG and provides some improvement of muscle force. PMID- 8239959 TI - Reliability of isokinetic muscle strength testing in 45- to 78-year-old men and women. AB - The test-retest reliability of isokinetic muscle strength testing was determined in 45- to 78-year-old men (N = 71; mean age = 60.2 years) and women (N = 107; mean age = 60.0 years). The dynamic muscle strength of the knee and elbow extensor (KE, EE) and flexor (KF, EF) muscle groups was measured at slow (60 degrees/sec) and fast (K = 240 degrees/sec; E = 180 degrees/sec) angular velocities using a Cybex II isokinetic dynamometer on two occasions seven to ten days apart. The mean peak torque in the second test was significantly higher (p < 0.001) in all muscle groups and both angular velocities tested in both sexes. The angle at which peak torque occurred did not change from the first to the second test except in the KF of females tested at 60 degrees/sec. Pearson's correlation coefficients (CCs; p < 0.01) in males ranged from .68 to .77 in the knee and from .71 to .84 in the elbow. In females, the CCs (p < 0.01) ranged from .58 to .74 in the knee and from .67 to .78 in the elbow. Thus in a large number of older men and women at least two tests may be necessary to determine isokinetic peak torque. PMID- 8239960 TI - Diagnosis and treatment of iliopsoas abscess in spinal cord injury patients. AB - Six patients with spinal cord injury and iliopsoas abscess and other complicating conditions were evaluated with computed tomography (CT), conventional radiography, magnetic resonance imaging (MRI), and radionuclide scans. CT identified the presence of psoas abscess and revealed the depth, extent, and relationship of deep pressure ulcers to deep structures. CT-guided aspiration of the abscess cavities was performed in three patients, with placement of drainage catheters. Concurrent treatment with appropriate antibiotics, followed by staged myocutaneous flap coverage resulted successful outcomes in all patients. A high index of suspicion aids in the early diagnosis of psoas abscess in the SCI patient, as interpretations of physical examination are obscured by the lack of localizing findings. We believe that CT is the diagnostic and therapeutic modality of choice in the management of these complex conditions in the SCI patient, because of its superior ability to detect pathologic changes in the pelvic region and for decreasing the morbidity of the treatment by avoiding open surgery in these often suboptimal surgical candidates. PMID- 8239961 TI - Aerobic exercise effect on individuals with physical disabilities. AB - The effects of aerobic exercise on the fitness level of adults with physical disabilities were examined in this research. Twenty-one individuals participated in a pretest-posttest research design that compared the effects of an aerobic exercise program upon experimental and control subjects, specifically, on body weight, resting, and exercise cardiorespiratory and metabolic parameters, and physical work capacity. Results indicate that aerobic exercise significantly improved physical work capacity by 23.1% and cardiorespiratory fitness as measured by maximal pulmonary ventilation and maximal oxygen consumption by 16.1% and 23.7% respectively, for the experimental group. In contrast, the control group demonstrated declines in these variables of 10.3%, 8%, and 17.1%, respectively. Findings from this research highlight the importance of exercise in promoting fitness and preventing secondary disabilities in adults with physical disabilities. PMID- 8239962 TI - Etiology, incidence, and prevention of deep vein thrombosis in acute spinal cord injury. AB - This article provides a critical review of the literature on the etiology, incidence, and prevention of deep-vein thrombosis in acute spinal cord injured patients. Stasis and hypercoagulability are the two major factors contributing to the development of thrombosis in this patient population. This has been supported by studies that demonstrate an impaired venous return from the lower extremities and abnormal coagulation factors, which predispose to thrombogenesis. The incidence of deep vein thrombosis secondary to the above etiologies varies from 49% to 100% in the first 12 weeks with the first 2 weeks having the highest rate following acute injury. This high rate of complication has led to numerous studies to identify the most effective regimens of prophylaxis. Studies using noninvasive testing and venography in acute spinal cord injury have supported two approaches for preventing deep-vein thrombosis. Single agent pharmacologic therapy with adjusted dose heparin is effective but does carry some risk of bleeding. Combination therapy with external pneumatic compression sleeves plus either aspirin/dipyridamole or low-dose heparin and electrical stimulation plus low-dose heparin have significantly reduced the incidence of deep vein thrombosis. The duration of prophylaxis with the above modalities has varied between 8 and 12 weeks following acute injury. Further large scale studies are required in this high-risk population to better delineate the incidence of deep vein thrombosis and pulmonary embolism, to identify the best modalities, and to define the duration of treatment for the prevention of these complications.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239963 TI - Clonidine-induced bradycardia in patients with spinal cord injury. AB - Clonidine is being used increasingly for treatment of spasticity in patients with spinal cord injury. Though hypotension, dry mouth, and constipation are well documented possible adverse effects, the possibility of clonidine-induced bradycardia is less well recognized and is rare. This report describes a patient who developed spasticity following a traumatic spinal cord injury. After clonidine was initiated, the patient's spasticity improved. However, he developed significant bradycardia. Once clonidine was discontinued, the resting heart rate returned to normal. This case illustrates an unusual adverse effect of clonidine. Possible mechanisms by which clonidine decreases spasticity are described, probable mechanisms of induced bradycardia are reviewed, and specific treatment recommendations for the use of clonidine in spinal cord injured patients are presented. PMID- 8239964 TI - Pulmonary embolism presenting as supraventricular tachycardia in paraplegia: a case report. AB - Supraventricular tachycardia as a presenting sign of pulmonary embolism is unusual. A 31-year-old man with incomplete T11 paraplegia on the basis of spinal cord ischemia developed supraventricular tachycardia 37 days after surgery to repair an aortic tear. Subsequent work-up revealed evidence of multiple submassive pulmonary emboli, despite thromboembolism prophylaxis. Clinicians should maintain a high index of suspicion for thromboembolic events when faced with cardiac rhythm disturbances in high-risk patients. PMID- 8239965 TI - Delayed femoral nerve palsy following femoral vessel catheterization. AB - We report two cases with a delayed complication of femoral vessel catheterization: femoral nerve entrapment by iliacus hematoma. Both patients had femoral artery catheterization and postoprocedural anticoagulation, followed by iliacus hematoma eruption on the side of catheterization. Diagnosis was confirmed by computed tomography and electrodiagnostic studies. The pathogenesis of iliacus hematoma-induced femoral neuropathy has been studied in detail; however, the authors could find no previous reports of catheterization causing this phenomenon. The association is probably under reported because of the delayed manifestation. Awareness of this complication could (1) minimize the neuropathy, by insuring early diagnosis and treatment and (2) reduce its incidence, by avoiding trauma to the iliacus muscle during catheterization. PMID- 8239966 TI - Distal humeral epiphyseal separation in a young athlete: a case report. AB - Elbow pain in the young throwing athlete can have numerous etiologies. Skeletally immature athletes commonly sustain epiphyseal injuries secondary to trauma and/or repetitive activity. The case of a 7-year-old Little League pitcher with the diagnosis of distal humeral epiphyseal separation is presented along with a review of the current literature. PMID- 8239967 TI - Chronic syndrome of inappropriate secretion of antidiuretic hormone in a pediatric patient after traumatic brain injury. AB - Chronic syndrome of inappropriate secretion of antidiuretic hormone (chronic SIADH) has been reported in adults after traumatic brain injury (TBI) but few similar cases have been reported in the pediatric population. We present a case of a 14-year-old boy who developed chronic SIADH after TBI in which caloric needs could not be adequately provided, with concomitant fluid restriction as the first line of treatment. Demeclocycline was ultimately used, which allowed for increased fluid liberalization and for provision of adequate calories. This form of therapy should be considered early in a child's course to prevent nutritional decline. Demeclocycline may be used for prolonged periods until the child's clinical condition permits fluid restriction to be effective therapy. PMID- 8239968 TI - Bladder augmentation using the stomach in spinal cord injured patients with impaired renal function. AB - A small capacity, poorly compliant neurogenic bladder is a difficult problem in spinal cord injured patients. Bladder augmentation with intermittent catheterization offers an alternative to indwelling catheterization. Ileum, cecum, and sigmoid colon have been most commonly used for enterocystoplasty but may, however, cause hyperchloremic metabolic acidosis in patients with impaired renal function. This can be prevented by performing a gastric augmentation. We present two cases of gastrocystoplasty in spinal cord injured patients with a small capacity, poorly compliant neurogenic bladder, impaired renal function, vesicoureteral reflux, and recurrent febrile urinary tract infections. Both patients had bilateral ureteral reimplantations during the surgery. One of the patients, a women, had a destroyed urethral sphincter secondary to long-term Foley that was repaired with a concurrent pubovaginal sling. Follow-up ranges from 12 to 18 months and both patients are continent with intermittent catheterization and have bladder capacities over 500 mL. Neither patient had deterioration in renal function or changes in serum electrolytes. Both patients maintain a slightly acidic urine and neither patient has had a clinically apparent urinary tract infection. PMID- 8239969 TI - Comprehensive analysis of energy storing prosthetic feet: Flex Foot and Seattle Foot Versus Standard SACH foot. AB - This study compared the mechanical and biomechanical functions, metabolic demand, and shock absorption of two dynamic elastic response (DER) prosthetic foot designs with the SACH foot. Nine individuals who had undergone unilateral below knee amputation were studied. Mechanical properties of the feet were related to gait biomechanics. Forefoot compliance is greatest for the Flex Foot and least for the SACH foot, hence, Flex Foot demonstrates (1) the longest midstance phase, (2) the greatest ankle angle range, and (3) greater forward movement of the center of pressure. There was some evidence that the DER feet produced a better push-off. However, neither the self-selected walking speed nor the metabolic rate or efficiency over a spectrum of walking speeds (73 to 120m/min) and running speeds (140 to 200m/min) was significantly different. Because no energy savings resulted for the DER feet, the release of stored energy in the flexible feet may not occur at the proper time to assist in ambulation as a result of the natural frequency of oscillation. PMID- 8239970 TI - Caveats in diagnosing protein S deficiency. PMID- 8239971 TI - Double-blind placebo crossover study of cyproterone acetate in the treatment of the paraphilias. AB - Cyproterone acetate is a powerful antiandrogen used in the treatment of the paraphilias for at least a decade. Studies have reported it effective in reducing the recidivism rates of sexual crimes perpetrated by men. It acts through competitive inhibition of the androgen receptors blocking the effects of testosterone and dihydrotestosterone. Systematic clinical study of its effectiveness and its clinical effects on sexual behavior is lacking, however. In the current study 19 paraphilic men who had also recidivated in a variety of sexual crimes were treated with cyproterone acetate and placebo in a double-blind crossover design. The use of active drug was associated with a significant reduction of some aspects of sexual behavior, particularly sexual fantasies. There were also significant effects on levels of circulating sex hormones and some effects on physiological measurements and self-reports of sexual arousal. PMID- 8239972 TI - Female sexual arousal and the law of initial value: assessment at several phases of the menstrual cycle. AB - Genital responses to erotic film and fantasy were recorded from 10 women at five phases of the menstrual cycle by means of vaginal photoplethysmography. Relationships between baseline (prestimulation) and difference (stimulation minus prestimulation) levels of response were examined. Neither vaginal blood volume nor average pulse amplitude conformed to the law of initial value ("the higher the baseline level, the less will be the difference score"). Baseline levels and difference scores generally were positively correlated at each phase of the menstrual cycle in the case of average pulse amplitude. For blood volume the correlations between baseline levels and difference scores were inconsistent in direction and small in magnitude, but trends differed between phases of the menstrual cycle. It is recommended that measures of female sexual arousal obtained through testing women who are at different phases of the cycle should not be aggregated unless it is established that the relationship between baseline levels and difference scores is uniform across phases. PMID- 8239973 TI - The effects of selective evaluation on the perception of female cues in sexually coercive and noncoercive males. AB - Various models have been proposed by which to understand the phenomenon of sexual coercion within dating relationships. Two are reviewed, including the limitations of each. A third, the situational approach, is outlined as more comprehensive in understanding the problem. One aspect of the model, selective evaluation, is experimentally tested. One-hundred eighty-two college students participated in brief heterosexual interactions and rated each other's behavior in terms of sexual expressiveness, flattery, and interest in future interactions. Behavioral data were also collected to evaluate differences between groups and the role behavioral cues play in the expression and attribution of sexual interest. Results support the hypothesis of selective evaluation in men, and in sexually coercive men in particular. Results are discussed with regard to socialization processes, communication of sexual interest, and recommendations for future research. PMID- 8239974 TI - Body image and sexuality in oophorectomized women. AB - Five groups of women ages 35 to 55 years were studied, including a nonsurgical control group (CTL), a hysterectomy-only (HYS), and three oophorectomy groups: an untreated group (UNT), women on estrogen replacement therapy (EST), and women on androgen-estrogen replacement therapy (COM). The interview/questionnaire session assessed mood, body image, and sexual functioning. In a second session completed by 58 of 129 subjects (45%), a photoplethysmograph measured vaginal blood flow in response to an erotic stimulus while subjects concurrently monitored subjective arousal. Overall, the UNT and EST groups had significantly lower self-reported desire and arousal than the remaining three groups. Body image was significantly poorer in the UNT group. All surgical groups reported more sexual problems than the control group. Furthermore, about a third of the CTL group reported positive changes in body image and sexuality in the previous 5 years. This effect was attenuated in the HYS, COM, and EST groups and almost absent in the UNT group. No significant group differences were obtained, however, on mood or vaginal blood flow and subjective arousal to an erotic stimulus. The role of differential levels of testosterone on sexuality is discussed as well as its clinical implications. PMID- 8239975 TI - Maternally rated childhood gender nonconformity in homosexuals and heterosexuals. AB - Homosexual and heterosexual subjects provided self-ratings of childhood gender nonconformity. Additionally, their mother rated them on several adjectives describing childhood behavior, which included words related to gender nonconformity. Male homosexuals were remembered by their mothers as less masculine and more nonathletic. This finding did not appear to be due to a bias in mothers' memories. Though female homosexuals were recalled as more masculine than female heterosexuals, this appeared to reflect retrospective bias, as mothers who knew of their daughters' homosexuality were more likely to rate them as masculine. Both self-rated and maternally rated childhood gender nonconformity made independent contributions in predicting sexual orientation. Within the homosexual samples, maternal and self-ratings of subjects' childhood gender nonconformity failed to correlate significantly. PMID- 8239976 TI - Increased vaginal blood flow induced by implant electrical stimulation of sacral anterior roots in the conscious woman: a case study. AB - The sacral anterior nerve roots were stimulated in a conscious female paraplegic by means of an intradural implanted Finetech/Brindley stimulator activated by inductive radio signals. Changes in vaginal blood flow were monitored by a photoplethysmograph. Stimulating S2 and S3 (but not S4) caused significant increases in vaginal pulse amplitude indicating genital vasodilation and increased blood flow. PMID- 8239977 TI - Effects of an oral contraceptive on sexual behavior of chimpanzees (Pan troglodytes) AB - Behavioral and physiological effects of a combined oral contraceptive (OC) were studied in chimpanzees for comparative purposes related to (i) the ambiguity surrounding the effects of OCs on the sexuality of humans, (ii) the close biological relationship between chimpanzees and humans, especially with respect to endogenous sex hormones and sexual behavior, and (iii) the relatively greater behavioral sensitivity of the chimpanzee to changes in sex hormone levels such as those that accompany the use of OCs. Two different types of pair tests and detailed behavioral assessments were used to differentiate the hormonal effects of female behavior from social effects imposed by the male. Anogenital swelling and copulation were reduced during OC cycles, but the effect of the OC on copulation was directly related to the social and sexual relationship of the pair during natural cycles. The more compatible and more frequently copulating pairs in the natural cycles continued to copulate during the OC cycles, albeit at reduced rates, whereas the less compatible and less frequently copulating pairs ceased copulating altogether when the female received the OC. There was no independent effect of the OC on ejaculation per se. Both male and female sexual initiative were reduced in the OC cycles, but female responsiveness to male sexual solicitations and direct copulatory attempts were not affected. Data indicate that despite generally adverse effects of the OC on certain physiological and behavioral dimensions of sexuality, social factors ultimately determined the degree of response to the OC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239978 TI - Carefully controlled population survey. PMID- 8239979 TI - Surgeons and molecular biology: a happy cohabitation. PMID- 8239980 TI - Molecular mechanisms of resistance to tamoxifen therapy in breast cancer. AB - Clinical data suggest that the use of adjuvant tamoxifen citrate (Nolvadex) for a minimum of 5 years, and possibly indefinitely, will result in maximal antitumor benefit. There is concern that long-term tamoxifen maintenance therapy may result in the induction of drug resistance. This article reviews the potential molecular mechanisms of resistance to tamoxifen and explores the possibility of tamoxifen stimulated tumor growth. PMID- 8239981 TI - Eicosanoids. Critical agents in the physiological process and cellular injury. AB - Eicosanoids were discovered as "prostaglandins" in the mid-1930s. The discovery that eicosanoids were ubiquitous in mammalian cells and that nonsteroidal anti inflammatory drugs worked by inhibiting enzymes that synthesized these chemicals heralded their extensive investigation in all fields of biology. Precursor fatty acids (arachidonic acids) are stored in cell phospholipids, acted on by two enzymes (cyclooxygenase and lipooxygenase) that yield prostaglandins, thromboxane, prostacyclin, and leukotrienes. Knowledge of their biochemical processes continue to unfold, but it is now believed that eicosanoids are part of a larger group of agents termed phospholipid mediators. Eicosanoids are intimately involved with cardiovascular function as well as central and peripheral vascular disease processes and ischemia. In the gastrointestinal tract, these potent lipids not only participate in many normal functions (eg, acid secretion and motility) but also in disease states (eg, inflammatory bowel disease and peptic ulcer disease). In shocklike states of sepsis and/or endotoxemia, eicosanoids have assumed a major role in many events that occur. Recently, discoveries have demonstrated that platelet-activating and tumor necrosis factors exert their effects in part through eicosanoids. The future will demonstrate these compounds to be critical not only in intracellular (molecular) events but also in the effects they produce that are far from the source of origin. PMID- 8239982 TI - Gene delivery systems in surgery. AB - Increased understanding of the genetic basis of human disease has led to a number of potential gene-based therapies for various medical and surgical disorders. The development of efficient methods for delivering genes to mammalian cells in vitro has increased the potential clinical utility of gene-based therapies; however, a major focus of research has been more efficient delivery to appropriate target cells, in vivo as well as in vitro, to establish gene therapy as an effective clinical modality for common disorders. Despite substantial progress, a number of critical technical issues to enhance and optimize not only gene transfer but also gene expression must be resolved. These future technological developments will be essential for the widespread clinical implementation of gene-based therapy. PMID- 8239983 TI - Cardiac preconditioning. Induction of endogenous tolerance to ischemia reperfusion injury. AB - Cardiac preconditioning is a phenomenon by which a brief exposure to ischemia renders the heart more tolerant of a subsequent sustained ischemic insult. Understanding the mechanism involved may allow pharmacologic access to this protective state before cardiopulmonary bypass surgery and transplantation. We discuss herein the preconditioning phenomenon, the potential mechanisms involved, and the therapeutic implications of cardiac preconditioning. PMID- 8239984 TI - Complement receptors in shock and transplantation. AB - Over the last decade, elegant studies of the basic biologic characteristics of inflammation and tissue injury have implicated leukocyte-mediated vascular and tissue injury in the pathogenesis of a wide variety of immune and inflammatory clinical disorders, including allograft rejection, adult respiratory distress syndrome, and shock. Recognition of the importance of leukocyte adherence to the endothelium in the pathogenesis of these disorders, in combination with advances in cellular and molecular biology, have led to the development of novel therapeutic approaches to the treatment of immune and inflammatory disorders in which leukocytes contribute to vascular and tissue injury. Several of these promising new therapeutic approaches have focused on the complement system. Examples of this therapeutic approach include the use of monoclonal antibodies directed at leukocyte complement receptors and the administration of soluble complement receptors to inhibit the binding of leukocytes to the endothelium. Because of the biologic and significant potential clinical importance of these advances, in this review, we focus on the complement system and complement receptor-mediated tissue injury. PMID- 8239985 TI - Growth factors. Local and systemic. AB - Growth factors that are produced locally at the wound site and systemically from distant sources may elicit inhibitory or stimulatory responses depending on interactions with other factors and the cellular environment into which they are liberated. Growth factors released from macrophages, neutrophils, lymphocytes, platelets, and fibroblasts bind to target cells via specific cell-surface receptors inducing cells to migrate, divide, or produce other factors required for wound healing. Structural similarities between cell receptors may explain cross-reactivity between substances. Systemic growth factors, such as growth hormone and local epidermal growth factor, fibroblast growth factor, platelet derived growth factor, and transforming growth factor beta have all been shown to enhance wound healing. Further, growth factors have been shown to have a distinct role in catabolic and malnourished patients as well as in patients with cancer. Clinical trials with growth factors have demonstrated efficacy, and specific uses for those particular factors may become an accepted mode of therapy producing important differences in outcome. PMID- 8239986 TI - Cytokine mediators of immunity and inflammation. AB - The remarkable advances in molecular and cell biology occurring over the past four decades have served the cause of surgical science well. Our understanding of basic disease mechanisms and insights into potential new therapeutic strategies have occurred at a staggering pace. Perhaps nowhere in surgical biology are these mechanistic insights and therapeutic prospects more evident than in research defining the cytokine mediators of inflammation, injury, and repair. These proteins are secreted to some degree by virtually all immune cell types as well as by a diverse array of other nucleated cells, and their functions encompass a regulatory role on and among many components of the immune system. Such intense interest is well deserved because abnormalities or dysregulation of tissue and wound repair as well as of natural (innate) or specific (acquired) immune function underlie much of the morbidity and mortality associated with surgical practice. Indeed, it is evident that the insights gained from the study of such inflammatory mediators cross virtually every specialty of surgery, from the acute sequelae of severe injury and invasive infection to the chronic manifestations of benign and malignant processes. PMID- 8239987 TI - Skin replacements. The biotechnological quest for optimal wound closure. AB - Extensive skin loss from a variety of conditions is associated with significant functional morbidity and loss of life. In many patients, a limited number of donor sites available for harvesting autologous split-thickness skin grafts prevents early, effective, and permanent wound closure. In the past 25 years, significant biotechnological advancements have been made in defining the criteria and manufacturing ingredients in materials that could serve as skin replacements for permanent wound closure. The optimal skin replacement should have the functional and cosmetic properties of the dermis and the epidermis. It should provide rapid, functional wound coverage and barrier protection to microorganisms, normalize fluid flux and hypermetabolism, and provide long-term stability without contraction or hypertrophic scarring. In addition, the optimal skin replacement should be nontoxic, easily stored and used, and relatively cost effective. This report will discuss the two major skin replacement designs available today, cultured keratinocyte grafts and bioartificial bilaminate systems, outline the advantages and disadvantages of each material, report the results of clinical trials for each, and speculate on the potential for each material to serve as a practical skin replacement. PMID- 8239988 TI - Clinical diagnostic applications of the polymerase chain reaction. Infection and rejection. AB - In this review, we describe the power and sensitivity of the polymerase chain reaction and indicate areas of clinical medicine in which it is currently being applied. As this technology and automation improves and is simplified, its realm of application will expand, but its major impact will continue to be its facilitation of early, specific diagnoses in infectious disease, genetics, cancer, and transplantation. Polymerase chain reaction is currently most useful in the care of opportunistic infections in immunosuppressed hosts and in making the distinction between infection and rejection in transplant recipients. As the technique becomes more widely available and less costly, its application should minimize the need for broad-spectrum antibiotic therapy of infections, lower costs, and perhaps even shorten hospitalization. These potential beneficial effects of polymerase chain reaction will be particularly relevant in the current cost-conscious health-care environment. PMID- 8239989 TI - Leukocyte responses to injury. AB - Injury elicits a response from all cells of the immune system in which cytokines and other metabolic products of activated leukocytes can act either beneficially to provide for enhanced host resistance or deleteriously to depress the function of remote organs and cause what has been termed systemic inflammation. These at times antithecal responses of leukocytes that appear to integrate postinjury changes in the neuroendocrine, immune, and coagulation systems have been implicated as principal causative factors in multiple systems organ failure. Numerous investigators have evaluated a variety of therapeutic agents to prevent and control infection by restoring leukocyte function, while others have evaluated antagonists and monoclonal antibodies as a means of controlling the exaggerated and persistent actions of leukocytes and cytokines caused by systemic inflammation. The redundancies of the cell populations and the cytokines and other metabolites produced by the cells predictably limit the effectiveness of any single agent and make clinical evaluation of such agents difficult. PMID- 8239990 TI - Biomolecular advances in gastrointestinal hormones. AB - Gastrointestinal hormones are chemical messengers that regulate a broad range of physiologic functions. Although primarily expressed within tissues of the gut, these peptide hormones are widely distributed throughout the body and act on multiple target tissues. Furthermore, these regulatory peptides can exist in multiple molecular forms that may bind to multiple cell-surface receptors coupled to one of several possible signal transduction systems leading to diverse biologic responses. With such an expansive field to study, it is not surprising that gut endocrinologists have embraced the new techniques that are emerging from the revolution of molecular biology. Beginning with the first construction of a recombinant DNA molecule by Paul Berg in 1971, molecular biology has developed many new techniques that have been rapidly adopted by gut endocrinologists to enable a more detailed understanding of gastrointestinal function. The merging of these two fields has led to a new area of research, molecular gut endocrinology, or the study of gut physiology and endocrinology at the level of individual molecules (ranging from polypeptide-surface receptors to small-molecule second messengers to DNA sequences). Gut cells are constantly bombarded by numerous hormones, and the tightly regulated physiologic status of each cell is becoming more clearly understood. PMID- 8239991 TI - Monoclonal antibodies for diagnosis and treatment. AB - One of the marvels of the host immune response is its response to antigenic foreign substances by manufacturing proteins that bind tenaciously to their targets. These proteins are antibodies or immunoglobulins produced in vast diversity during an individual's lifetime. By virtue of this process, the mammalian host possesses the innate ability to mount an initial response to antigens to which there has been no prior experience and to develop an even more effective response on reexposure to these same substances. This capacity to distinguish self from nonself is one of the most basic aspects of the cellular and humoral arms of the immune response and is one of the primary means by which the host combats infection caused by many different types of pathogens. In this context, antibodies have long been recognized as a critical component of host defenses and are capable of binding to invading microbes and microbial toxins. PMID- 8239992 TI - Adoptive cellular therapy of malignancy. AB - The realization that human cancers can be responsive to the manipulation of the immune system has only recently been documented. The immune approaches to the treatment of malignancy can be broadly classified into either active or passive immunotherapies. With active immunotherapy, treatment relies on the in vivo stimulation of the endogenous host immune system to react against tumors with the administration of biological response-modifying agents (ie, bacterial adjuvants, cytokines, tumor vaccines). With passive immunotherapy, treatment involves the delivery of biologic reagents with established tumor-immune reactivity (ie, antibodies or cells) that can directly or indirectly mediate antitumor effects and does not necessarily depend on an intact host immune system. Cellular therapy of malignancy has become more feasible with increased understanding of the interactions between immune cells and tumors. This article will review our current understanding of the principles underlying these interactions. PMID- 8239993 TI - Toxicokinetic interactions between chlorinated aromatic hydrocarbons in the liver of the C57BL/6J mouse: I. Polychlorinated biphenyls (PCBs). AB - 2,2',4,4',5,5'- (PCB 153), 2,3,3',4,4',5- (PCB 156) and 3,3',4,4',5,5' hexachlorobiphenyl (PCB 169) were administered orally to three groups of C57BL/6J mice using single doses of 1.5-109.1 mg/kg. Two other groups of mice received binary mixtures of PCB 153 and 156 or PCB 153 and 169. The hepatic deposition, elimination, CYP1a and CYP2b dependent enzyme activities were studied during a 77 day period. Some interactive effects on hepatic deposition and elimination were observed, resulting in increased deposition and faster elimination. These effects were most pronounced for the PCBs 156 and 169. A potentiating effect on hepatic CYP1a dependent 7-ethoxyresorufin-O- deethylation (EROD) activity was observed for the combination of PCB 156 and 153. Based on the results from the present study and earlier studies, it is suggested that the potentiating effect on EROD activity might be caused by a mechanism that is governed by at least two factors. The first is a toxicokinetic modulation of hepatic retention. The second factor is probably an elevation of hepatic Ah receptor levels by PCB 153. PMID- 8239994 TI - Pathways of trans,trans-muconaldehyde metabolism in mouse liver cytosol: reversibility of monoreductive metabolism and formation of end products. AB - The metabolism of trans,trans-muconaldehyde (MUC), a hematotoxic agent which is a presumed in vivo metabolite of benzene, was studied in mouse liver cytosol. MUC was incubated for 30 min at 37 degrees C with mouse liver cytosol (from CD-1 mice) supplemented with NAD+ and the products were analyzed by reverse phase HPLC. Two products were detected in addition to the previously identified acid aldehyde 6-oxo-trans,trans-2,4-hexadienoic acid (COOH-M-CHO) and the diacid trans,trans-muconic acid (COOH-M-COOH). Based on the molecular weight (112) obtained by thermo-spray LC-mass spectrometry and the absorbance maximum (269 nm), one of the products was identified as the aldehyde-alcohol 6-hydroxy trans,trans-2,4-hexadienal (CHO-M-OH). The second product was identified as 6 hydroxy-trans,trans-2,4-hexadienoic acid (COOH-M-OH) by coelution with authentic standard, the fragmentation pattern obtained by electron impact mass spectrometry and the absorbance maximum (258 nm). Time course and concentration dependency studies indicate that COOH-M-OH and COOH-M-COOH are end products of MUC metabolism while CHO-M-OH, and COOH-M-CHO, the initially formed mono-reduction and mono-oxidation products, respectively, are the intermediates leading to these end products. The metabolite COOH-M-OH is formed mainly by oxidation of CHO-M-OH and to a much lesser extent by reduction of CHO-M-COOH, whereas COOH-M-COOH is formed solely by oxidation of COOH-M-CHO. The reduction of MUC to CHO-M-OH is reversible, whereas oxidation to COOH-M-CHO is not. The compound CHO-M-OH is not only oxidized to COOH-M-OH by oxidation of the aldehyde functional group, but is also converted back to MUC by oxidation of the alcohol functional group. PMID- 8239995 TI - Measurement of hemoglobin and albumin adducts of benzo(a)pyrenediolepoxide and their rate of elimination in the female Sprague-Dawley rat. AB - A high performance liquid chromatographic technique coupled with fluorometric detection was used to study the disappearance rate of the hemoglobin and albumin adducts of benzo(a)pyrene (BaP) in female Sprague-Dawley rats. The technique was considered adequate based on good recovery (> 80%) of the analyte and satisfactory intra-assay coefficients of variations of 7.8% and 4.2% for hemoglobin and albumin adducts, respectively. The persistence of the adducts was determined in rats injected intravenously with 550 nmol BaP diolepoxide, the electrophilic metabolite of BaP. In a first experiment the isolated protein fractions were directly subjected to mild acid hydrolysis to yield the ultimate analyte, BaP tetrol. In a second experiment these fractions were first digested by proteases in order to liberate any unbound lipophilic BaP tetrol possibly "hidden" in the hydrophobic portions of the protein matrices. It was found that 70-100% of the measured tetrols originated from true adducts. The half-life of the free tetrol was 2.8 and 1.6 days in the hemoglobin and albumin fractions, respectively, compared to 10.7 and 3.6 days for the true adducts. Based on the total amount of tetrols, the mean half-life of the latter in the hemoglobin and albumin fractions determined from the two experiments was 9.0 and 2.7 days, respectively, assuming first order kinetics. The greater persistence of the hemoglobin adduct compared to that of the albumin adduct is coherent with the biological half-life of the intact proteins. However, the observed disappearance profile obtained does not fit with current models of adduct formation and removal. PMID- 8239996 TI - Enzyme induction by ethanol consumption affects the pharmacokinetics of inhaled m xylene only at high levels of exposure. AB - The experimental study with rats was undertaken to verify the working hypothesis that enzyme induction caused by ethanol consumption affects the kinetics of m xylene only at a high level of exposure. m-Xylene was administered to ethanol treated rats either perorally (0.01, 0.02 or 0.1 ml/kg) or by inhalation (50, 100 or 500 ppm each for 6 h) and the concentration of m-xylene in the blood and the urinary excretion of a m-xylene metabolite (m-methyl hippuric acid or m-MHA) were measured with time. The ethanol consumption, which increased the in vitro m xylene metabolism about 5-fold, had no effect on the metabolism of inhaled m xylene in vivo until the exposure concentration was raised to 500 ppm. On the other hand, metabolism of m-xylene after oral administration was markedly enhanced at any dose by the consumption, as evidenced by a decrease in the blood concentration of m-xylene together with an increase in the urinary excretion of m MHA. These findings indicate that enzyme induction does not affect the pharmacokinetics of inhaled m-xylene when its exposure concentration is low. This may be because the hepatic blood flow, rather than the enzyme activity, rate limits the metabolism of m-xylene, which is highly metabolized in the liver. PMID- 8239997 TI - Characteristic modifications of the breathing pattern of mice to evaluate the effects of airborne chemicals on the respiratory tract. AB - A system was developed for exposure of unanesthetized mice to airborne chemicals and for continuous measurement of their breathing pattern prior to, during and following exposure. By measuring inspiratory and expiratory airflows (VI and VE), and integration with time to yield tidal volume (VT), we obtained characteristic modifications to the normal breathing pattern. These permitted recognition that a specific portion of the respiratory tract was affected by the selected airborne chemicals. Following recognition, we also quantitated the degree of effect using one specific measurement in each case. An effect on the upper respiratory tract, induced by the sensory irritant, 2-chlorobenzylchloride, was quantitated by measuring a decrease in respiratory frequency. An effect on the conducting airways, induced by the airway constrictor, carbamylcholine, was quantitated by a decrease in VE at the mid-point of VT. An effect at the alveolar level, induced either by the vagal nerve ending stimulant, propranolol, or by the pulmonary irritant, machining fluid G, was quantitated by an increase in the length of a pause induced at the end of expiration. The system is easy to construct and operate and can be used to rapidly evaluate the effects of airborne chemicals on the respiratory tract. PMID- 8239998 TI - Neuron and glial cell marker proteins as indicators of heavy metal-induced neurotoxicity in neuroblastoma and glioma cell lines. AB - The sensitive and specific biochemical indicators for assessing chemical-induced neurotoxic insults in cell culture models have not been sufficiently explored. This study was designed to assess the usefulness of glia-specific beta-S100 protein and neuron-specific enolase (NSE) as indices of in vitro neurotoxicity of heavy metals. Glioma C6 and neuroblastoma N18TG-2 cells were grown in Dulbecco's modified Eagle's medium containing various concentrations of mercuric chloride (HgCl2) or cadmium chloride (CdCl2) for 5 days. Toxic response patterns of the neurospecific endpoints (beta-S100 and NSE), which were monitored with enzyme immunoassays, were compared with those of the non-neurospecific endpoints such as cell viability, total cellular protein, lactate dehydrogenase (LDH) activity, and cumulative glucose consumption in the two cell lines. Both HgCl2 and CdCl2 produced dose-dependent inhibition of neurospecific endpoints and non-specific endpoints. However, by ranking the EC50 values (effective concentration producing half-maximal inhibition) for various endpoints, the lowest values were found for beta-S100 in C6 cells, and for NSE in N18TG-2 cells. In lower and intermediate concentrations, the inhibitory effects of the heavy metals on the content of beta S100 and NSE occurred in the absence of any detectable effect on intracellular LDH activity, and independently of total cellular protein inhibition. The sensitive and excess responses of the neurospecific endpoints relative to that of the non-specific endpoints may reflect the specific neurotoxic insults of the heavy metals on the cultured cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8239999 TI - Possible involvement of active oxygen species in selenite toxicity in isolated rat hepatocytes. AB - Mechanisms of selenite cytotoxicity were examined using isolated rat hepatocytes. When selenite was added to a suspension of rat hepatocytes, intracellular reduced glutathione (GSH) was decreased and the oxygen consumption rate was increased. Subsequently, thiobarbituric acid-reactive substances (TBA-RS) and lactate dehydrogenase (LDH) leakage were increased. A ferric iron chelator, desferrioxamine (DF), and a synthetic superoxide dismutase (SOD) mimic, desferrioxamine manganese (DFMn), reduced the selenite toxicity. These results suggest that superoxide anion and its reactive metabolites such as the hydroxyl radical may be involved in the cytotoxicity of selenite. PMID- 8240000 TI - Investigation of the renal injury caused by liver ischemia-reperfusion in rats. AB - To explain the mechanism of renal injury caused by liver ischemia-reperfusion, we investigated biochemical and morphological changes in the liver and kidney in rats. After reperfusion following 60 min of liver ischemia, numerous changes were found. The level of serum transaminases and lipid peroxide formation in the liver tissue increased significantly. Electron microscopic studies revealed that most of the hepatocytes had swollen mitochondria and clumping of the nuclear chromatin. The sinusoidal endothelium was disrupted and the sinusoidal lumen was filled with numerous erythrocytes. Blood endotoxin concentration, plasma lipid peroxide levels, and serum beta-glucuronidase activities were significantly higher than in the control group. Biochemical and morphological renal injury was also observed. Tissue lipid peroxide levels increased in both the kidney and the liver. Microscopic examination revealed damage to the renal tubules, including interstitial edema, dilatation of the lumen, and granular casts derived from necrotic cells in the proximal convoluted tubule. The levels of urinary N-acetyl beta-D-glucosaminidase (NAG) in the liver ischemia-reperfusion group were also higher than in the control group. These results suggest that the renal injury was caused by an increase in endotoxin, lipid peroxide, and lysosomal enzymes in the blood following the liver injury induced by the ischemia-reperfusion. PMID- 8240001 TI - Acute toxicity of two pyrethroids, permethrin, and cypermethrin in neonatal and adult rats. AB - The present study aims specifically at obtaining a comparison of the acute toxicity of cypermethrin (CY), a type I pyrethroid, and permethrin (PERM), a type II pyrethroid, administered orally as a single dose to neonatal and adult rats, and at assessing the importance of pyrethroid biotransformation in CY and PERM toxicity through use of drug metabolism inhibitors. Our experiments show that CY is more toxic than PERM to adult and neonatal rats. The sensitivity of neonatal rats both to CY and to PERM toxicity is higher, the younger the animals. CY is much more toxic than PERM in the neonatal rat, compared with the adult. In rats aged 8, 16, and 21 days, pretreatment with piperonyl butoxide (PB), a monooxygenase inhibitor, or with tri-o-tolyl phosphate (TOTP), an esterase inhibitor, does not produce significant variations in the lethal effects of CY and PERM. Instead, in the adult rats, a significant increase in CY (chi 2 = 5.97; p < 0.05) and PERM (chi 2 = 4.37; p < 0.05) mortality occurred in rats pretreated with esterase inhibitors, whereas no increase in CY and PERM toxicity was found in adult animals pretreated with monooxygenase inhibitor. It was concluded that the higher level of sensitivity of the neonate rat to pyrethroid toxicity is probably due to incomplete development of the enzymes which catalyze the metabolism of pyrethroids in the liver of young animals. It is suggested that ester hydrolysis is an important pyrethroids detoxification reaction in the adult rat. PMID- 8240002 TI - Histamine and 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline are competitive inhibitors of debrisoquine 4-monooxygenase in rat liver. AB - The effects of histamine and 1-methyl-6,7-dihydroxy-1,2,3,4- tetrahydroisoquinoline(salsolinol) on debrisoquine 4-monooxygenation, which is catalyzed by cytochrome P-450db 1 (CYP2D1) in rat liver microsomes, were studied. Both histamine and salsolinol competitively inhibited the activity of debrisoquine 4-monooxygenase (Ki = 0.31 and 0.43 mM, respectively). These data demonstrate that histamine and salsolinol bind to the active site of CYP2D1, i.e. histamine and salsolinol have structures (molecular shape) corresponding to the active site of CYP2D1. PMID- 8240003 TI - Analysis of host genetic control of scrapie-induced obesity. AB - The potential for induction of obesity during the preclinical phase of scrapie disease in mice was previously shown to be a function of both the strain of scrapie and the strain of inbred mouse. In the present study, host control of obesity induction by a scrapie strain was examined to determine if the effect were dependent on a single gene or multiple genes. The approach used was assessment of the pattern of weight induction in F1 and F2 crosses of parental inbred mouse strains that did or did not show a weight increase with a specific scrapie strain. Analyses of these data indicated that the induction of obesity was controlled by multiple host genes. In an unrelated observation, there was a correlation between the incubation period of a strain of scrapie in F2 generation mice and their coat color, i.e., the average incubation period of yellow-brown mice was significantly less than those of either black or white mice. PMID- 8240004 TI - Genetic and biological differentiation of dengue 3 isolates obtained from clinical cases in Java, Indonesia, 1976-1978. AB - Previous epidemiological, virological and clinical studies have documented a series of outbreaks of dengue fever and dengue haemorrhagic fever/dengue shock syndrome which occurred in Java, Indonesia in 1976-1978. In the current study we compare growth characteristics in cell culture, and nucleotide sequence data for the viral prM and E genes, of five low passage DEN-3 isolates obtained during these epidemics from clinically defined cases. All isolates had the same passage history: human sera were passed twice in mosquitoes and three times in a mosquito cell line (Aedes albopictus, C 6/36 cells). Growth differences were observed between individual isolates in Vero cells; growth differences were not observed in C 6/36 cells. Nucleotide sequencing of the prM and E gene region indicated that no two isolates were identical (sequence divergence ranged from 0.4 to 1.6% in pairwise comparisons) but that they were closely enough related to present a single genetic type. There were one or two differences in deduced amino acid sequence in E between isolates. Differences were at residues 65, 187, 298 or 443. One isolate differed from all others at residue 16 in the M protein. No relationship was apparent between the amino acid sequence of M or E and the nature of the disease profile, the year of isolation or the geographic region of isolation. The isolates showed 3.5 to 4.4% nucleotide sequence divergence from the highly-adapted H 87 prototype, isolated in the Philippines in 1956. The isolates showed a total of twelve common amino acid differences in prM and E proteins from H 87. Ten of these twelve residues were at positions which differed between the four dengue serotypes. Two differences (at residues 37 in M and 293 in E) were at positions which are conserved in sequence between the four dengue serotypes. The data are discussed in relation to the dengue outbreaks in Java in the period 1976-1978. PMID- 8240005 TI - On the relationship between X-bodies and symptom development in plants infected with different tobamoviruses. AB - The relationship between systemic mosaic symptoms and the occurrence of viral 126 kDa protein in X-bodies was studied in tobacco infected with the tobacco mild green mosaic virus (TMGMV) strains U2, U5, and ribgrass mosaic virus (RMV) strain HR, and in other plant species infected with tobacco mosaic virus (TMV) strain W U 1. Strains U2, U5, and HR coded for proteins of 126, 126, and 130 kDa, respectively, but these were not recognized by antisera against the corresponding protein from W U 1. Only the HR 130-kDa protein reacted with an antiserum raised against a peptide of amino acids 849-863 from the sequence of W U 1. Electron microscopic analysis established the presence of virus clusters in the cytoplasm, as well as in chloroplasts, in leaf tissue infected with U 2 or U 5, and adjacent to nuclei and chloroplasts in scattered cells infected with HR. X-bodies were not detected after infection with any of these strains, but were large and adjacent to nuclei in W U 1-infected tomato displaying severe mosaic symptoms. Large X bodies were detected near nuclei in W U 1-infected tomato displaying severe mosaic symptoms, but none were detected after infection of tobacco with any of the other tobamoviruses. The induction of X-bodies appears to be characteristic of some tobamovirus only and, at best, can only be associated with, rather than causative of, the severity of symptoms induced by those viruses. PMID- 8240006 TI - Use of NS3 consensus primers for the polymerase chain reaction amplification and sequencing of dengue viruses and other flaviviruses. AB - Consensus primers for the polymerase chain reaction were designed based on conserved motifs within the serine protease and RNA helicase domains encoded by the NS 3 genes of dengue and other flaviviruses. Target fragments of 470 bp were amplified on cDNA templates synthesized from RNAs of dengue types 1, 2, 3, and 4, Japanese encephalitis, Kunjin, and yellow fever viruses using random or specific downstream primers. PCR of oligo(dT)-primed cDNAs from Japanese encephalitis and Kunjin viral RNAs did not yield target bands. As few as 10(3) copies of dengue viral RNA could be detected. Direct DNA sequencing of PCR products of reference strains of dengue 2 (NGC), Kunjin (MRM 61C) and yellow fever (17 D) viruses demonstrated complete concurrence with published data. However, 2 nucleotide differences were observed between our data for dengue 3 H87 strain and the published sequence, resulting in a single amino acid disparity. Differences at 21, 16, and 11 nucleotide positions were noted between dengue 1 Hawaii and S 275/90; dengue 4 H 241 and 814669; Japanese encephalitis Nakayama and JaOArS 982 viral strains, culminating in only 4, 1 and 1 amino acid residue differences, respectively. These amino acid disparities occurred outside putative active sites of the enzymatic domains, emphasizing the important role of the NS3 protein in flaviviral replication. This RNA-PCR consensus primer strategy coupled with DNA sequencing represents a valuable tool for the molecular diagnosis and epidemiology of dengue and other flaviviral infections. PMID- 8240007 TI - DNA sequence of a gene cluster in the equine herpesvirus-4 genome which contains a newly identified herpesvirus gene encoding a membrane protein. AB - Complete DNA sequences for the equine herpesvirus-4 (EHV-4) genes analogous to equine herpesvirus-1 (EHV-1) genes 8, 9, 10, and 11, varicella zoster virus (VZV) genes 7, 8, 9 A, and 9, and herpes simplex virus type 1 (HSV-1) genes UL51, UL50, UL49A, and UL49 are presented. The EHV-4 gene corresponding to EHV-1 gene 10/VZV gene 9A/HSV-1 UL49A is of particular interest in that it is a newly identified herpesvirus gene whose product demonstrates features characteristic of membrane inserted proteins. Furthermore, this gene has counterparts in all herpesvirus genomes sequenced to date. PMID- 8240008 TI - The role of the immune system in establishment of herpes simplex virus latency- studies using CD4+ T-cell depleted mice. AB - The immunological mechanisms involved in establishment of herpes simplex virus (HSV) latency were studied in normal and CD4+ T-cell depleted C57BL/6J mice following intravaginal infection. During transition from acute to latent ganglionic infection two consecutive processes were observed: first, clearance of infectious virus from the ganglia, and second, reduction of the number of infected ganglia. PMID- 8240009 TI - Isolation of a virus with rhabdovirus morphology from a white-beaked dolphin (Lagenorhynchus albirostris). AB - A virus with rhabdovirus morphology which proved to be antigenically distinct from rabies virus and vesicular stomatitis virus was isolated from a dolphin that had beached on the Dutch coast. Neutralizing antibodies to this virus were found in several European marine mammal species. PMID- 8240010 TI - The porcine paramyxovirus LPM specifically recognizes sialyl (alpha 2,3) lactose containing structures. AB - The porcine paramyxovirus LPM recognizes alpha, but not beta, anomers of sialic acid containing structures, specifically sialyl (alpha 2,3) lactose. The virus specificity is directed to the sialyl residue and to the C'4 axial OH and the C'6 CH2OH of the galactose present in this structure. PMID- 8240011 TI - The mumps virus V protein is unstable in virus infected cells. AB - The mumps virus (MuV) V protein was characterized in virus infected cells by the use of antipeptide sera. In radioimmune precipitation assay (RIPA), the sera reacted with the V protein and also immunoprecipitated the nucleocapsid (NP) and phospho (P) proteins. However, by depletion RIPA (in which either the NP and P proteins or the V protein were removed) and Western immunoblotting, it was demonstrated that the V protein was not associated with the NP and P proteins, but that the anti-V sera cross-reacted with the NP protein. Pulse-chase experiments demonstrated that the V protein was gradually decreased during the chase period and could not be detected by antibodies raised against peptides representing three different regions of the protein at the end of the chase, while the NP and P proteins were relatively stable during the chase period. These results suggest that the V protein is unstable and degraded gradually in virus infected cells. PMID- 8240012 TI - Biosynthesis and morphogenesis of group C rotavirus in swine testicular cells. AB - Polypeptide synthesis and morphogenesis of a group C rotavirus (AmC-1) adapted to a continuous swine testicular cell line was examined. SDS-PAGE analysis of 35S methionine labeled infected cell lysates revealed 9 viral polypeptides (122, 98, 79, 78, 43, 41, 35, 24, and 20 kD). Viral polypeptide synthesis appeared to be maximal at 7-10h post infection. Purified group C virus grown in the presence of trypsin was found to contain seven structural polypeptides (122, 98, 79, 53, 43, 41, and 30 kD) by protein blotting and five polypeptides (98, 79, 78, 43, and 41 kD) by immunoprecipitation with a hyperimmune rabbit antisera. Tunicamycin treatment, Concanavalin A binding, protein blotting, endo-H treatment and 2,6H mannose labeling suggested that group C rotavirus contains one structural glycoprotein (41 kD) with a corresponding precursor mol. wt. of 37 kD and one not previously identified nonstructural glycoprotein (24 kD) with a corresponding precursor mol. wt. of < or = 20 kD. Electron microscopy of infected swine testicular cells revealed an assembly process for group C rotavirus similar to group A, with single-shelled particles budding through the rough endoplasmic reticulum with concomitant acquisition of a transient membrane. PMID- 8240013 TI - Raccoon poxvirus rabies virus glycoprotein recombinant vaccine in sheep. AB - Twenty sheep were divided into groups and inoculated by various routes with recombinant raccoon poxvirus expressing the CVS rabies virus glycoprotein (rRCNV G) or with raccoon poxvirus (RCNV). The apparent innocuous pathologic responses to each virus coupled with development of high levels of rabies virus neutralizing antibodies in animals vaccinated with rRCNV-G intradermally or intramuscularly suggested that the recombinant is effective and that RCNV would be a suitable substrate for further development of sheep vaccines. Poor antibody response to rRCNV-G given orally implied that it would be relatively harmless if inadvertently ingested by sheep. Virus transmission between vaccinated and sentinel sheep was not observed or detected serologically. PMID- 8240014 TI - Nucleotide sequence and E. coli expression of the coat protein gene of the yellowing strain of soybean dwarf luteovirus. AB - The nucleotide sequence of the coat protein gene of the yellowing (Y) strain of soybean dwarf virus (SbDV) was determined from cloned cDNA. The gene contains 600 nucleotides and encodes a protein of 200 amino acids with a calculated M(r) of 22,200. The identity of the open reading frame (ORF) encoding the coat protein was confirmed by expression of an Escherichia coli beta-galactosidase fusion protein and detection by a dot blot immunoassay. Sequence comparisons of the deduced coat protein amino acids to several luteoviruses demonstrated that the SbDV-Y coat protein ORF shares greatest similarity with bean leaf roll virus (BLRV) (65%). PMID- 8240015 TI - Genetic and antigenic analysis of an equine influenza H3 isolate from the 1989 epidemic. PMID- 8240016 TI - Comparative analysis of the genomes of mycobacteriophages infecting saprophytic and pathogenic mycobacteria. AB - The genomes of a series of mycobacteriophages have been analyzed to disclose possible relationships between genetic characteristics and host range. The percent guanine-plus-cytosine in the DNA of 14 phages was found to be 34.4 to 47.5, as determined by a double-labelling procedure, which is unaffected by the presence of modified bases. The DNA of few mycobacteriophages yielded discordant values when the G + C content was estimated by buoyant density determination and by the double labelling procedure. This observation suggests the possible presence of modified bases in these genomes. The reduced susceptibility of viral DNAs to several restriction endonucleases is suggestive of the occurrence of both methyladenine and methylcytosine in the genome of all the mycobacteriophages studied. Heterologous annealing among the 14 DNAs analyzed yielded 6 hybridization groups. Within one group, the homology level among viral genomes was estimated by comparing the electrophoretic mobilities of restriction fragments: values of 0.8 to 1.3% base substitution have thus been found. A comparison of the genomic characteristics and host range of the mycobacteriophages analyzed suggests a possible relationship between restriction pattern, G + C content, crosshybridization level and host range. PMID- 8240017 TI - Extensive sequence variation of feline immunodeficiency virus env genes in isolates from naturally infected cats. AB - In an investigation of the evolution of feline immunodeficiency virus (FIV) in vivo, sequential isolates from a persistently infected cat were examined by direct sequencing following amplification of selected subgenomic regions by polymerase chain reaction (PCR). Three isolates, T90, T91, and T92, obtained over a three-year period revealed no changes to regions known to be conserved within gag and pol genes. Additionally, no change occurred within gag and pol in an isolate recovered from a second cat which was experimentally infected with T90. Changes were detected within an N-terminal region of the envelope glycoprotein gp 120 (env). These consisted of point mutations, some of which would result in amino acid substitutions and the predicted amino acid changes tended to cluster within variable domains. Inoculation of T90 into a second cat resulted in a different pattern of mutations than that observed for the three isolates from the first cat. In all cases, virus isolates derived from the same cat were much more highly related to each other (extent of env variation was 0.5-1.5%) than to isolates from other cats (10-12% env variation). The rate of change of FIV was estimated to be 3.4 x 10(-3) nucleotide substitutions per site per year for the env gene and less than 10(-4) nucleotide substitutions per site per year for the gag and pol genes, values concordant with that found for human immunodeficiency virus 1. Both nucleotide and amino acid changes in the gp 120 region were found to be directional, suggesting that selective pressures influence FIV envelope gene sequences. PMID- 8240018 TI - Detection and characterization of a novel bisegmented double-stranded RNA virus (picobirnavirus) from rabbit faeces. AB - In two separate studies rabbits were fed orally with human and rabbit "picobirnaviruses". Polyacrylamide gel electrophoresis (PAGE) of nucleic acid extracted from faecal samples collected from inoculated rabbits revealed the presence of discrete equimolar bands, typical of picobirnaviruses, in several specimens. The genome profiles detected in both studies differed significantly from that of the inoculum suggesting that passage of the inoculated picobirnaviruses had not taken place and that the bands were a co-incidental finding. The presence of rabbit picobirnaviruses was confirmed by characterization of the genome bands, as dsRNA by enzyme digestion and by their co-sedimentation in caesium chloride (CsCl) gradients with 32 nm virus particles at a buoyant density of 1.39 g/ml. Picobirnavirus genome segments varied in size in a range between 2.3-2.6 kilo base pairs (kbp) and 1.6-1.9 kbp for the slow and fast migrating bands, respectively. Immune electron microscopy of the picobirnavirus particles revealed round or slightly hexagonal particles with a smooth surface and a mean diameter of 30.7 nm. In one rabbit, an immune response, temporally associated with picobirnaviruses excretion, was demonstrated by immune electron microscopy (IEM) supporting the view that picobirnaviruses may be vertebrate viruses. Two antigenically distinct picobirnavirus strains were defined by IEM. PMID- 8240019 TI - A study of five bacteriophages of the Myoviridae family which replicate on different gram-positive bacteria. AB - A comparative study is reported on five phages of the Myoviridae family which propagate on Bacillus subtilis, B. thuringiensis, Enterococcus sp., Lactobacillus plantarum, or Staphylococcus aureus. The phages are morphologically identical and characterized by isometric heads with conspicuous capsomers and by contractile tails with complex base plates. The phages show similar protein profiles, but vary considerably in burst size. Phage DNAs are about 95-166 kb in size and are unrelated by DNA-DNA hybridization and restriction endonuclease analysis. Therefore the phages are unrelated at species level. Implications of these data for our understanding of the development of phage species are discussed. PMID- 8240020 TI - Expression of two bovine rotavirus non-structural proteins (NSP2, NSP3) in the baculovirus system and production of monoclonal antibodies directed against the expressed proteins. AB - Studies on rotavirus non-structural proteins have been hampered in the past by difficulties in obtaining monospecific reagents. To make such reagents available, we have expressed in the baculovirus system NSP2 and NSP3 (formerly called NS35 and NS34, respectively) of the bovine rotavirus RF and produced hybridomas against these proteins. Full-length DNA copies of RNA segments 7 (coding for NSP3) and 8 (coding for NSP2) of the virus strain RF were cloned and sequenced. Each cDNA was inserted in the transfer vector pVL941 and used to transfect Spodoptera frugiperda cells (Sf9). Recombinant baculoviruses encoding these proteins were obtained. Infection of Sf9 cells with these recombinant viruses resulted in a high level of expression of NSP2 and NSP3 (range of 1 microgram per 10(6) cells). Monoclonal antibodies (MAbs) were elicited by immunization of BALB/c mice with adjuvented, unpurified recombinant proteins in the rear foot pads. Fusion was performed using lymphocytes from popliteal lymph nodes with SP2/O-Ag14 myeloma line. Screening was by differential indirect immunofluorescent staining on monolayers of Sf9 cells infected with each recombinant virus. Two MAbs proved to be reactive against NSP3 and a single one against NSP2. They showed high specificity by immunofluorescence, immunoprecipitation and Western blot. The isotype of these MAbs was IgG1. Oligomeric forms of NSP3 and NSP2 proteins were detected and the existence of intra-chain disulfide bridge in NSP2 protein was suggested. The levels of synthesis and cellular localization of NSP3 and NSP2 proteins were different as shown by immunoprecipitation and immunofluorescence. PMID- 8240021 TI - Identification of 108K, 93K, and 42K glycoproteins of bovine herpesvirus-1 by monoclonal antibodies. AB - Three glycoproteins of bovine herpesvirus-1 (BHV-1) other than glycoproteins gI, gIII, and gIV were identified by monoclonal antibody (MAb) analyses. Monoclonal antibodies were obtained by immunization of mice with either BHV-1 envelope or virus infected cells, from which the glycoproteins gI, gIII, and gIV were removed by immunoaffinity. In the latter immunization procedure mice were tolerized either against normal cellular antigens with or without glycoproteins gI, gIII, gIV, and nucleocapsid. From 154 anti-BHV-1 hybridomas isolated, 39 MAbs precipitated a 108K glycoprotein. Two other glycoproteins of respectively 42K and 93K were precipitated each by one MAb. These three glycoproteins were detected in infected cell lysate. Nine anti-108K glycoprotein MAbs neutralized BHV-1 infectivity and three non-neutralizing MAbs were able to reduce plaque development when virus was grown in the presence of these MAbs. It is therefore suggested that this glycoprotein is involved in viral entry into the cell and in cell-to-cell spread of the virus. PMID- 8240022 TI - Diagnostic value of finger thermometry in assessment of hand-arm vibration syndrome. AB - Finger thermometry and cold provocation test were performed in 90 chain saw workers and 58 healthy controls. The workers were grouped according to the Taylor Pelmear scale of hand-arm vibration syndrome into subgroups 0, 1, 2, 3. The recovery rate reflected passive vasodilative ability. According to the results of the study, the test could differentiate the subjects with clinically manifest vibration-induced white fingers (stages 1, 2 and 3) from the controls. The discriminating threshold of recovery rate with regard to Raynaud's phenomenon was 90%. With that diagnostic limit the above method was sensitive and specific enough to verify vibration-induced vasospastic disorders and therefore could be recommended for their surveillance. PMID- 8240023 TI - A new method for determining extra time by considering ergonomic loads in the garment and metal-working industries. AB - The changing labour conditions in the garment and metal-working industries have led to the necessity of determining new extra times to establish the time standard. In this paper, a method of measuring stress and strain imposed upon the operator in new working conditions by determining the additional production coefficient is presented. The method gives criteria and grades to assess stress at the workplace. Physical stress (dynamic and static), thermal and visual stress, discomfort caused by noise, aerosols, gases and vapours, and the stress due to monotony are assessed. The stresses are expressed by ecological, physiological and psychological indexes. The indexes are pondered. Ponderation includes the factor of the magnitude and the time of exposure to stress. Ponderation is expressed by the number of points, the total being an adequate presentation of the stresses. Stress and strain decrease the working performance of the operator. Therefore they must be included in the standard to compensate for the operator's lower working efficiency due to physical and psychological strain at the workplace. Short breaks and stops allowed during work enable the operator to perform work tasks without getting excessively tired. The necessary correction of the production time is made by applying the ergonomic coefficient Ker. PMID- 8240024 TI - Fatal poisonings in the city of Zagreb. AB - The paper deals with the results of analysis of fatal poisonings in the city of Zagreb from 1981 to 1990. The work is an extension of earlier analyses for the same area that were carried out from the foundation of the Department for Forensic Medicine and Criminology, School of Medicine, University of Zagreb in 1934 to 1967, and from 1968 to 1980. Comparative analysis of all data shows that despite the increasing rate of poisoning the number of cases of fatal poisoning has been steadily diminishing--from 11 cases per 100,000 population in the 1934 1967 period and eight cases between 1968 and 1980 to five cases in the latest period 1981-1990. PMID- 8240025 TI - A field-test for detecting organophosphorus compounds in water. AB - An enzyme test has been worked out for detecting organophosphorus compounds in water. The test is based on the inhibition of cholinesterase. The detection limits for the "nerve gases" are (microgram/L): Soman 0.12, VX 5.9, Sarin 9.9 and Tabun 26. The detection limit for the organophosphorus pesticide dichlorvos is 50 micrograms/L. PMID- 8240026 TI - [Detection of occupational lead poisoning and specific permanent work disability]. AB - Evaluation of occupational lead poisoning and resultant permanent work disability calls for a procedure which is not easy to conduct. This is because vague, non manifest lead poisonings and transient pre-poisoning states have lately prevailed over clearly manifested, obvious lead poisonings as were found earlier due to unfavourable working conditions. A reason for this further lies in non-specific clinical symptoms of lead poisoning as well as in lack of a strict set of rules- especially when it comes to assessing biological indicators of abnormal lead absorption in the body--for experts in the disability assessment boards to go by. In view of that the conduct of those taking part in the evaluation procedure (general practitioner of patient's own choice and team of experts in pension and disability assessment boards) is analysed. Many assessments have been found deficient. An amendment to the rules for expert evaluation is proposed. PMID- 8240027 TI - Neomycin--a frequent contact allergen. AB - The incidence of allergic contact dermatitis from neomycin evaluated in relation to 1381 verified cases of allergic contact dermatitis showed a progressive increase (5.00, 7.69, 10.18%) over a three-year period (1990-1992). Sensitivity to neomycin was investigated with special reference to possible cross-reactions between neomycin and the allergens that are commonly used in the manufacture of cosmetic products. Contact sensitivity to neomycin was found to be present with the other diagnoses, such as atopic dermatitis, seborrhoeic dermatitis, hypostasic dermatitis and psoriasis vulgaris. PMID- 8240028 TI - [Respiratory function in confectionary workers]. AB - Respiratory symptoms and ventilatory capacity were studied in 259 female confectionary workers and in 65 non-exposed controls. Most of the chronic respiratory symptoms appeared more frequently among the exposed workers, particularly if they worked in exposure to flour, talc, starch, acids and alcohol. There was a high prevalence of acute symptoms that developed during work shift. This was true especially of cough, dyspnea, burning and dryness of the throat and eye irritation. The exposed workers exhibited statistically significant acute reductions of ventilatory capacity. Those were particularly pronounced for FEF50 (range: 4.6-13.0%) and FEF25 (range: 4.7-22.3%). The measured preshift values of ventilatory capacity were significantly lower than the predicted normal values. The administration of disodium chromoglycate (DSCG 40 mg) significantly diminished mean across-shift reductions in all ventilatory capacity tests. Data suggest that sensitive confectionery workers may develop acute and chronic respiratory symptoms accompanied by acute and/or chronic changes in ventilatory capacity. PMID- 8240029 TI - [Aging and work capability]. AB - With the average life duration becoming longer there is an increasing number of old people in the population. In the literature the problem of old people is still neglected. The most visible changes in old people occur in physical characteristics and functions; at first a change in psychological functions was also reported to take place. However, it was shown that results depended to a great extent on the method used: the longitudinal method (following up one and the same generation) frequently gave rather different and much better results than the cross-sectional method. The question of old age and working abilities is an important one. Certain positive personality traits seem to compensate for a drop in certain abilities. In the modern automated industry time pressure and adaptation to new working conditions are a handicap for the elderly, but the application of different ergonomic principles can greatly improve the situation. PMID- 8240030 TI - Effect of cholesterol oxides on prostacyclin production and platelet adhesion. AB - Prostacyclin (PGI2) is synthesized primarily by endothelial cells, is essential for maintenance of vascular integrity, and may play a role in atherogenesis. Human umbilical vein endothelial cells in culture were incubated with either pure cholesterol, 25-hydroxycholesterol, 7-ketocholesterol, cholesterol 5 alpha,6 alpha-epoxide or cholestane-3 beta,5 alpha,6 beta-triol at 10 micrograms/ml culture medium concentration for 12 hours and 24 hours. PGI2 production measured by radioimmunoassay of 6-keto PGF1 alpha, the stable metabolite of PGI2 was inhibited by 39.6%, 27.3%, 40.1% and 31.9% after incubation with 25 hydroxycholesterol, 7-ketocholesterol, cholesterol 5 alpha, 6 alpha-epoxide or cholestane-3 beta,5 alpha,6 beta-triol for 12 hours respectively. Further inhibitory effects were shown after 24 hours of incubation with 25 hydroxycholesterol and 7-ketocholesterol. Platelet adhesion onto endothelial cell monolayers measured by 111In-labeled platelets was enhanced by 104%, 54% and 37% after incubation with cholestane-3 beta, 5 alpha,6 beta-triol, 25 hydroxycholesterol, and 7-ketocholesterol at 10 micrograms/ml concentration for 12 hours respectively. Pure cholesterol at the same concentration had no effect on PGI2 production or platelet adhesion. PMID- 8240031 TI - Effect of aspirin on the contractility of aortic rings in vitro from spontaneously hypertensive rats. AB - The effect of Acetylsalicylic acid (ASA) on the responsiveness of rat aortic smooth muscle was investigated by comparing the contractility of aortic rings from SHR and WKY rats in response to alpha-agonist, phenylephrine (PE). Cumulative dose response curves for PE were generated from aortic rings of SHR and WKY rats in the presence and absence of 0.2mM ASA. To investigate the involvement of endothelium on ASA mediated effects, active tension was recorded for denuded and non-denuded rings from SHR animals in response to KCl and PE. Denuded and non-denuded aortic rings from SHR animals in the presence of ASA produced significantly higher active tension than in the absence of ASA. The reactivity of aortic rings from WKY control animals was not altered significantly in the presence of ASA. These studies suggest that ASA can also modulate aortic contractility through other mechanism(s) in addition to its effect on the metabolites of arachidonic acid. Significance of these observations is discussed below. PMID- 8240032 TI - Human parathyroid hormone dilates both pig coronary and human inferior epigastric arteries by a cyclic AMP-dependent pathway. AB - Human parathyroid hormone (hPTH (1-38)) induced concentration-dependent relaxations in prostaglandin F2 alpha-preconstricted pig coronary arteries in vitro. Removal of endothelial cells or pretreatment with nitro-L-arginine, a specific inhibitor of the endothelium-derived relaxing factor (EDRF) synthesis, impaired, although to a small extent, hPTH-induced relaxations. Human PTH-, but not bradykinin- or nitroglycerin-induced relaxations were potentiated in the presence of the cyclic AMP phosphodiesterase inhibitor 3-isobutyl-1 methylxanthine (IBMX). Human PTH also relaxed preconstricted human inferior epigastric arteries in vitro. In accordance to pig coronary arteries, this relaxation was potentiated in the presence of IBMX. However, the human internal thoracic (mammary) artery did not respond to hPTH (1-100 nM). Thus, acute vasodilatory effects of hPTH may not be present in all human arteries. The physiological significance of this phenomenon is not known. This relaxation, at least in pig arteries, is mediated to a small extent by the release of EDRF. In addition, this relaxation appears to be mainly mediated in both pig and human arteries by a smooth muscle cyclic AMP pathway. PMID- 8240033 TI - Interactions between cultured bovine arterial endothelial and smooth muscle cells; further studies on the effects of injury and modification of the consequences of injury. AB - The hypothesis that cells of the arterial wall might modify the consequences of arterial injury was tested. Bovine aortic endothelial cells (EC) or smooth muscle cells (SMC) were exposed to the two toxic stimuli 3,4-benzo(a)pyrene (BP) and dimethylsulfoxide-soluble particulate matter from cigarette smoke (DSP) or factors released from platelets. The modification of the injury caused by these substances on arterial cells was studied by using a conditioned medium from arterial cells or an EC-SMC co-culture model. Direct addition of BP or DSP to the EC or SMC cultures induced toxic effects on the cells. DSP caused a decreased release of prostacyclin by EC. Conditioned medium from EC and SMC modified these toxic effects, which resulted in a reduced cell death and a further decreased cell proliferation, while conditioned medium from SMC increased the release of prostacyclin by EC injured by DSP. In EC-SMC co-culture the same modifications were obtained. The modification of cell injury was not linked to cell proliferation but instead the results suggested that the effects were mediated by multiple substances released from arterial cells. It is concluded that interactions between different cells in the arterial wall, in the non-injured as well as in the injured state, could be modified by endogeneous substances. This might be of relevance for atherogenesis. PMID- 8240034 TI - [Congestive heart failure. Concept of diastolic and systolic heart failure. A didactic presentation]. PMID- 8240035 TI - [Hemodynamic effects of low doses of chlorthalidone in patients with mild systemic arterial hypertension]. AB - PURPOSE: To evaluate the hemodynamic effects of low doses of chlorthalidone (CHT) in patients with systemic arterial hypertension (SAH). METHODS: Eight patients with mild SAH, mean age of 52 +/- 8.9 years, 7 men, were studied. Clinical evaluation, systolic (SBP) and diastolic (DBP) blood pressure and heart rate (HR), in supine and standing positions, were obtained before and every two weeks, first two in placebo, during 12 weeks. Laboratory data, hemogram, sodium, potassium, urea, creatinine, glucose, hepatic aminotransferases and urinalysis, were done and at end of study. Hemodynamic monitorization was performed by Swan Ganz catheter in pulmonary artery to obtain RAP and PAWP, in mmHg. Cardiac output (CO) was obtained by thermodilution method. Systemic vascular resistance (SVR) arose from variables above. Hemodynamic variables were measured at 2nd and 12th weeks during treatment with 50mg of chlorthalidone each 48 h. RESULTS: A significant reduction of SBP (p = 0.005 and p = 0.003), DBP (p < 0.0001 and p < 0.0001), respectively in supine and standing positions. HR did not show statistical difference. At hemodynamic monitoring was observed a significative reduction of SVR (p < 0.02), but not with CO. CONCLUSION: Chlorthalidone in low dosis was effective to treat mild SAH, basically by lowering SVR. PMID- 8240036 TI - [Symptomatic mitral valve prolapse. Left ventricular dysfunction and beneficial effect of digitalis]. AB - PURPOSE: To study the left ventricular function by M-mode echocardiography in symptomatic patients with primary mitral valve prolapse (MVP), without significant mitral regurgitation, during the phenylephrine test, before and after digitalization. METHODS: Thirty one patients with MVP and 10 normal subjects as a control group were studied by M mode echocardiography: Percentage of systolic shortening (DD), ejection fraction (EF) and mean velocity of circumferential fibers shortening (Vcf) were measured. The echocardiographic recordings were obtained at rest and during the continuous infusion of phenylephrine (40-60 micrograms/min) in order to allow an increase of 20-30 mmHg in systolic pressure. RESULTS: At rest, cardiac size and function were normal in both groups. During phenylephrine infusion in 27 patients a decrease in DD (from 37.7 +/- 4.6 to 31.0 +/- 4.0, p < 0.001); in EF (from 0.76 +/- 0.05 to 0.66 +/- 0.6, p < 0.001); in Vcf (from 1.05 +/- 0.77 to 0.76 +/- 0.13, p < 0.001) were observed. From this group 20 patients received digoxin. After the digitalization the phenylephrine test did not cause changes in DD (from 38.3 +/- 5.0 to 39.2 +/- 3.8, NS); in EF (from 0.77 +/- 0.06 to 0.77 +/- 0.04, NS); in Vcf (from 1.05 +/- 0.19 to 0.94 +/- 0.13, NS). These patients during the submaximal exercise test, showed significant increases at the total work load (from 617 +/- 248 to 982 +/- 244 watts, p < 0.001) and cardiac efficiency (from 25.0 +/- 11.5 to 37.2 +/- 10.4, p < 0.001). On the other hand the phenylephrine infusion in the control group did not result in changes in DD, EF and Vcf. CONCLUSION: Symptomatic patients with primary MVP showed total recovering of left ventricular dysfunction with digitalization during the phenylephrine test, with improvement of cardiac efficiency. PMID- 8240037 TI - [Infectious endocarditis in a drug addict]. AB - PURPOSE: To compare two groups of patients with infective endocarditis, the drug addicts and non-drug addicts. We attempted to set particularities among the various aspects that involve the patient with endocarditis, due to the concurrent chronic use of cocaine intravenously. METHODS: Twenty nine patients, group B, whose clinical diagnose was compatible with infective endocarditis, with risk factor of parenteral toxicomania by cocaine were treated at Institute "Dante Pazzanese de Cardiologia" and Hospital "Emilio Ribas" in Sao Paulo, from 1984 to 1990. The data obtained for etiological agents, previous cardiac pathology, affected heart structures, affected heart side and clinical-surgical evolutions of group B were compared to group A (193 patients), which was also composed of patients with endocarditis, without chronic endovenous use of cocaine antecedent. The data obtained were analysed comparatively according to the chi square with Yates correction. RESULTS: Male gender (89.7%) was predominate in group B towards group A (57.0%); (p < 0.01). Previous cardiopathy, either congenital or acquired, as antecedent proning to endocarditis, was found in 89.1% of patients in group A, significantly higher than 17.2% of patients group B (p < 0.001). Staphylococcus aureus was the most frequent agent, which accounted for endocarditis of group B in 86.4% of the cases, significantly higher when compared to 23.9% of cases of group A (p < 0.01). Streptococcus viridans was the most frequent etiological agent for endocarditis of group A (44.8%), significantly higher than group B (4.5%), (p < 0.01). In concern to the affected structures, the tricuspid valve was most affected in group B (65.5%), significantly higher than group A (4.7%) p < 0.001. The mitral valve was significantly more affected in group A (45.1%) in comparison to group B (6.9%), (p < 0.05). In group A 82 patients (42.5%) required surgical treatment and this occurred in 3 patients of group b (10.3%), (p < 0.05). No significant statistical difference was found as for the general mortality (clinical and surgical) in both groups. CONCLUSION: a) presence of previous cardiac disease was lower suggesting permanent contamination blood flow by pathologic agents, mainly of those found in the skin as S. aureus; b) right side of the heart is most frequently affected, specially the tricuspid valve even without previous damage. PMID- 8240038 TI - [Leukocyte activation during extracorporeal circulation. The next aim of knowledge]. PMID- 8240039 TI - [Regression of tachycardiomyopathy after implant of pacemaker with antitachycardial function]. AB - A 27-year-old woman with systemic lupus erythematosus and Wolff-Parkinson-White syndrome complicated with refractory tachycardia and class III heart failure treated with pacemaker implantation was described. She had cardiomyopathy that could be due to lupus erythematosus or tachycardia-induced. Nonpharmacologic therapeutic alternative was used and a universal DDD pulse generator with selected programming was chosen. Twenty-four months follow-up showed tachycardia control and regression of symptoms of heart failure to class I as well as improvement of left ventricular function evaluated by echocardiographic method. Thus, pacemaker implant may be an useful alternative approach in patients with tachycardiomyopathy in whose other nonpharmacologic therapeutic options could not be performed. PMID- 8240040 TI - [Double aortic arch associated with stenosis of left pulmonary artery]. AB - Case report on a 6-year-old patient with vascular ring, due to double aortic arch. The patient was asymptomatic. The barium-filled esophagus showed bilateral indentations and a large posterior indentation. Echocardiographic study in notch suprasternal view revealed features of this pathology. The diagnosis was made by the cineangiocardiographic study that showed double aortic arch with separate carotid and subclavian arteries arising from each arch. The right arch was larger than the left, and the descending aorta was on the left side. There was severe left pulmonary artery stenosis associated. The clinical management was chosen, because the patient was asymptomatic. PMID- 8240041 TI - [Ischemic cardiomyopathy in a child. Case report with accentuated coronary disease]. AB - Ischemic cardiomyopathy in childhood is related with congenital and acquired pathologies which could lead to serious cardiac sequelae as myocardial infarction and sudden death. Unfortunately, when coronary artery malformations are excluded, it is difficult in some cases to be completely certain on the etiology of the arterial coronary disease, due the lack of pathognomonic diagnostic tests. Case report of 6 year old child, with severe coronary artery disease: aneurysm of main branch of left coronary artery; left anterior descending branch and right coronary artery occlusions with accentuated myocardial dysfunction. We believe that the diagnosis of Kawasaki disease is presumptive in this patient. The principal aspects to elucidate the etiology of coronary artery disease are analysed, as well as the importance of identifying patients without known previous illness, but with severe coronary artery lesions, as in this reported case. PMID- 8240042 TI - [Cardiovascular dysfunction in septic shock and its treatment]. PMID- 8240043 TI - [Pregnancy in cardiovascular disease. Experience with 1000 cases]. PMID- 8240044 TI - [Unsupervised physical exercise for patients with heart diseases--"imperative"]. PMID- 8240045 TI - [Aortic insufficiency due to quadricuspid aortic valve]. AB - In a 45 year old patient with clinic diagnosis of aortic regurgitation the two dimensional echocardiography demonstrated a quadricuspid aortic valve and severe aortic regurgitation by Doppler. The quadricuspid aortic valve is a rare pathology, usually not associated with other cardiovascular pathologies and in which there is evolution with aortic regurgitation due to malposition of the four cusps. PMID- 8240046 TI - [Pulmonary vascular remodelling. Considerations regarding the possible participation of growth peptides factors and related substances]. PMID- 8240047 TI - [Ramipril, a new antagonist of angiotensin converting enzyme]. PMID- 8240048 TI - [Evaluation of efficacy of the enalapril-hydrochlorothiazide combination by ambulatory blood pressure monitoring in essential hypertensive patients]. AB - PURPOSE: Evaluation the 24 hours efficacy of once daily enalapril plus hydrochlorothiazide association by ambulatory blood pressure monitoring. METHODS: Thirty-nine essential hypertensive patients (3 male, 36 female; 31 white, 8 non white; mean age 46,7 years old) with the criteria, after wash-out period, of more than 40% of diurnal diastolic blood pressure measurements above 90mmHg by ambulatory blood pressure monitoring, were allocated for treatment during 8 weeks with once daily administration of enalapril (20mg) plus hydrochlorothiazide (12.5mg) association. RESULTS: After wash-out period, 82 and 42%, respectively, diurnal and nocturnal systolic blood pressure measurements were above 140mmHG; while diastolic values were 79 and 26% above 90mmHg. After 8 weeks of treatment there was a significant reduction in both systolic and diastolic pressure loads, either on nocturnal or diurnal periods; 26 and 5.3% of systolic values were still above 140mmHg and, 31.5 and 7.9% of diastolic measurements were above 90mmHg. Despite the significant fall on blood pressure there was not alteration in heart rate. CONCLUSION: The association of the angiotensin converting enzyme inhibitor, enalapril, plus a diuretic, hydrochlorothiazide, promoted a significant reduction on pressure load and did not interfere with the circadian rhythm of 24 hours blood pressure. These results may indicate that the association as suitable as one of the first choices for treating mild and moderate hypertensive patients. PMID- 8240049 TI - [First Brazilian consensus for the use of ambulatory blood pressure monitoring. Brazilian Society of Cardiology]. PMID- 8240050 TI - [Coronary disease. Epidemiologic and preventive aspects]. PMID- 8240051 TI - [Catheter ablation in patients with refractory cardiac arrhythmias with radiofrequency techniques]. AB - PURPOSE: evaluate the efficacy of radiofrequency catheter ablation in patients with refractory cardiac arrhythmias. METHODS: twenty patients with refractory cardiac arrhythmias were undertaken to electrophysiologic studies for diagnosis and radiofrequency catheter ablation of their reentrant arrhythmias. Ten patients were men and 10 women with ages varying from 13 to 76 years (mean = 42.4 years). Nineteen patients had supraventricular tachyarrhythmias: One patient had atrial tachycardia and 1 atrial fibrillation with rapid ventricular rate, 5 patients had reentrant nodal tachycardia, 12 patients had reentrant atrioventricular tachycardia and 1 patient had right ventricular outflow tract tachycardia. RESULTS: the mean time of the procedure was 4.1 hours. The radiofrequency current energy applied was 40-50 V for 30-40 seconds. Ablation was successful in 18/20 (90%) patients; in 15/18 (83%) of successfully treated patients the same study was done for diagnosis and radiofrequency ablation. One patient had femoral arterial occlusion and was treated with no significant sequelae. During a mean follow-up of 4 months no preexcitation or reentrant tachycardia occurred. CONCLUSIONS: the results of our experience with radiofrequency catheter ablation of cardiac arrhythmias suggest that this technique can benefit an important number of patients with cardiac arrhythmias. PMID- 8240052 TI - [Chagas disease and primary arterial hypertension]. AB - PURPOSE: To evaluate the pressoric behaviour in chagasic patients in different stages of myocardial damage and to correlate Chagas' disease with arterial hypertension. METHODS: The arterial blood pressure of 644 chagasic patients surveyed in the HC-UNICAMP, with arterial hypertension or not, associated with the stage of myocardial dysfunction was evaluated. This group was compared with 370 hypertensive patients without Chagas' disease. Both were divided by sex and age. The results were evaluated by covariance analysis and the significance was pointed at p < 0.05. RESULTS: The chagasic group presented a high prevalence of arterial hypertension with 16.8% in the clinical phase I, 27.8% in the phase II and 34% in the III. Both hypertensive and nonhypertensive chagasic patients had a significant drop in clinical phase I after the age of 30 years. The chagasic patients classified in phase II presented a high prevalence between 30-50 years, not associated with the pressoric behaviour and/or sex. The symptomatic myocardial dysfunction (phase III) was uncommon below 40 years old in chagasic group with hypertension or not, but presented a highest prevalence in male hypertensive chagasic patients above 50 years old. CONCLUSION: The study demonstrated a more usual association of Chagas' disease and hypertension in patients with symptomatic myocardial dysfunction, above 50 years old. This find evidenciates the somatory and progressive effect of both diseases. It is possible that the pathogenic mechanisms of Chagas' disease contributed to the increment of the arterial blood pressure. PMID- 8240053 TI - [Amiodarone in cardiac failure]. AB - PURPOSE: To analyse the effects of amiodarone on the treatment of cardiac failure. METHODS: The effects of 200 mg/day of amiodarone were assessed in 18 patients with a history of cardiac failure. Functional class, heart rate, blood pressure, left ventricular ejection fraction, heart size, treadmill exercise tolerance and electrocardiogram were assessed before and eight weeks after the use of amiodarone, and the side effects were monitored. RESULTS: In 18 patients receiving amiodarone, the functional class and heart rate decreased significantly (p < 0.05) from 2.7 +/- 0.4 to 1.2 +/- 0.4 and 103 +/- 15 to 69 +/- 8.5 bpm respectively. The ejection fraction and the exercise tolerance increased significantly (p < 0.05) from 29.2 +/- 5.5% to 41.5% and from 379.8 +/- 271 sec to 897.8 +/- 350.8 sec, respectively. The incidence of atrial and ventricular arrhythmias decreased significantly and no side effects were observed. CONCLUSION: Amiodarone appears to produce benefits in patients with cardiac failure with atrial and ventricular arrhythmias. PMID- 8240054 TI - [Left heart catheterization by the transseptal route]. AB - PURPOSE: To analyse the transseptal catheterization technique, its indications and possible complications. METHODS: The transseptal catheterization has been undertaken in 233 patients, 202 children (aged 0.1 to 16 years) with congenital heart disease and 31 adults (aged 37 to 73 years) submitted to mitral valvuloplasty. The Mullins technique was employed to access left heart chambers. RESULTS: In the pediatric group, the indications for transseptal catheterization were coarctation of the aorta and valvular or subvalvular aortic stenosis. By this technique, multiple diagnostic analysis and therapeutic procedures, such as blade atrioseptostomy and mitral valvuloplasty, became available. Among the complications in the 2 groups, pericardial perforation was the most frequent, depending on the laboratory practice with this technique. CONCLUSION: The transseptal cardiac catheterization is a safe and effective technique to investigate hemodynamic data in several congenital heart diseases and is essential to many therapeutic procedures. The technique has a low incidence of complications when employed by experienced teams. PMID- 8240055 TI - [Morphologic study of the inferior cavo-atrial transition in man. Morphofunctional considerations]. AB - PURPOSE: Analysis the fibrous elements of the transition between the inferior vena cava and right atrium. METHODS: Twenty adult (20-40 ys) were used. Properly preserved samples were analysed macro and microscopically. Fragments were excised from the inferior cavo-atrial transition and analysed under stereomicroscopic. RESULTS: The inferior vena cava valve was disposed at the wall of the vein. Insertions of striated muscular cardiac fibers bundles are seen at the anterior wall of inferior vena cava. These muscular bundles are directed towards the intima of the vein. There were at the venous posterior wall insertions of the cardiac muscular bundles, originated from the crista terminalis and from the interatrial septum, these bundles show an orientation predominantly oblique, with tendency to become circular. At the transition level, the bundles of smooth muscular fibers of the inferior vena cava, presented an oblique disposition, becoming predominantly circular. CONCLUSION: As the disposition of muscular cardiac fibers presents a predominantly circular direction and, the bundles of smooth muscular fibers of the inferior vena cava present the same direction, we could postulate that, when the atrial systole occurs, these muscular bundles would obliterate the cavo-atrial transition, preventing, this way, the venous reflux. PMID- 8240056 TI - [Prevalence of congestive heart failure after ventriculomectomy in patients with obstructive hypertrophic cardiomyopathy]. AB - PURPOSE: To assess the prevalence of congestive heart failure in the late post operative follow-up in patients with hypertrophic obstructive cardiomyopathy (HOCM) submitted to ventriculomectomy (VM). METHODS: Thirty six patients with HOCM, all of them with basal intraventricular systolic pressure gradient (BPG) > or = 50 mmHg, were studied. Sixteen (group I) were submitted to VM and 20 (group II) remained under clinical treatment. At group I the ages were 1 month to 61 years (28.25 +/- 15.39), 10 male (62.5%); BPG ranged from 50 to 192 mmHg (98.09 +/- 42.76) with mean follow-up of 13.06 +/- 7.19 years. At group II the age ranged from 2 months to 58 years (31.01 +/- 18.61); 12 male (60%); BPG ranged from 52 to 185 mmHg (83.47 +/- 35.08) with mean follow-up of 9.6 +/- 4.93 years. RESULTS: At group I the follow-up was good in 4 patients (25%), stable in 2 (12.5%) and poor in 3 (18%). Seven patients (43.75%) developed congestive heart failure at mean time of 7.53 +/- 8.44 years; there was one operative death and the follow-up control was lost in 7 patients. In group II, the follow-up was good in 3 patients (15%), stable in a 6 (30%) and poor in a 9 (45%); no deaths occurred. Two patients had heart failure within 3 years after the onset of the disease. The follow-up control was lost in 8 patients (40%). CONCLUSION: 1) congestive heart failure was more frequent in patients submitted to VM rather than the non-operated ones (p < 0.05); 2) the short-term follow-up was better in operated patients rather than in non-operated ones. PMID- 8240057 TI - [Heparin in coronary angioplasty. Randomized study in cases with low risk of acute occlusion]. AB - PURPOSE: To assess the efficacy of heparin in preventing the abrupt closure after coronary angioplasty in low risk patients for this phenomenon. METHODS: In the last 4 years, 525 patients successfully dilated were randomized to receive intravenous heparin (n = 264) or not (n = 261) after the angioplasty. The excluding criteria were contraindications for heparin and risk for abrupt closure (refractory unstable angina, primary coronary angioplasty in acute myocardial infarction, evidence of intracoronary thrombus, intimal tear after the procedure and cases of chronic total occlusions). Both heparin and non heparin groups were similar in respect to female sex (15% x 17%; p = NS), age over 70 years old (7% x 9%; p = NS), previous myocardial infarction (26% x 24%; p = NS), multi-vessel procedures (4% x 7%; p = NS, stable angina (40% x 46%; p = NS), unstable angina (52% x 48%; p = NS) and angioplasty after thrombolytic therapy (8% x 6%; p = NS). RESULTS: The overall incidence of abrupt closure was 2/525 (0.4%), with one case (0.4%) in each group. The in-hospital mortality was 1/525 (0.2%), which occurred in a non-heparin patient, due to a anterior myocardial infarction. Major complications occurred similarly in heparin and non-heparin groups (0.4%). Bleeding complications were observed more frequently in the heparin group (7% x 2%; p = 0.002). All of them were in the catheterization site and none required blood transfusion. Severe systemic bleeding were not observed. CONCLUSION: In patients regarded as low risk for abrupt closure, the incidence of this complication was really low (0.4%) and heparin probably do not prevent it. PMID- 8240058 TI - [Chagas disease--review of 8 simultaneous cases of acute Chagas myocarditis: 25 years later]. AB - PURPOSE: To analyse the evolution of 8 cases of Chagas' disease after a microepidemy of 17 cases which occurred in Teotonia-RS-in 1965 and to discuss the importance of oral contamination in Chagas' disease. METHODS: All 8 patients are followed at the IC/FUC-RS-Brazil since 1980. Clinical examinations, blood tests, ECG, Cx-Ray and Echo were made in 1991. RESULTS: None of the eight cases presented any sign of chronic Chagas' disease manifestation. The blood tests, ECG, Cx-Ray and Echo were normals. Only one case showed a immunofluorescence of 1/20. CONCLUSION: 1) The follow-up after 25 years in all the 8 cases did not show clinical alterations or any chronic manifestation of Chagas' disease. All the tests performed had normal results; 2) the oral transmission must be considered definitive in Chagas' disease. PMID- 8240059 TI - [Carcinoid syndrome]. PMID- 8240060 TI - [A comparative study of pancreatic-duodenal transplantation, islets of Langerhans transplantation, and insulin treatment, in the control of experimental diabetes in the rat]. AB - Sixty outbred Wistar rats were randomly assigned to five experimental groups: GI 10 non-diabetic control rats; GII-10 untreated diabetic control rats; GIII-10 diabetic rats treated with retard porcine insulin; GIV-20 diabetic rats that received pancreaticoduodenal transplantation (PDT) from normal donor rats; GV-10 diabetic rats submitted to islet of Langerhans transplantation (ILT) into the portal vein. The animals were housed in metabolic cages for six periods of 24 hours during 30 days and body weight, water and food intake, urine output, blood and urinary glucose were recorded. Diabetes was induced by I.V. administration of Alloxan (42 mg/kg of body weight); PDT was performed by microsurgical techniques and islets were prepared without enzymes. To prevent rejection. Cyclosporin A (10 mg/kg of body weight) was utilized in transplanted rats. PDT consistently and significantly (p < 0.05) improved the metabolic abnormalities of the diabetic rats, by restoring the body weight gain, and immediate relief of polydipsia, polyphagia, polyuria, hyperglycemia and glucosuria observed in pre-treatment period. PDT was more effective than ILT and this over insulin therapy on control of the diabetic state. However, the observed complications in GIV and GV, due to surgery and immunosuppression, should be analysed for the real benefits of the alternative therapy can be superior to eventual fails to the conventional therapy with insulin. PMID- 8240061 TI - [Primary non-Hodgkin's lymphoma of the liver: report of two cases and review of the literature]. AB - Two cases of primary non-Hodgkin lymphoma of the liver are presented, with comments about the main clinical and pathological aspects of the disease based on literature review. Both patients were woman (19 and 62 years old) and presented the same complaints: severe weight loss and presence of fast growing abdominal mass. In both cases the histologic diagnosis showed a non-Hodgkin, diffuse, large cell lymphoma (centroblastic-centrocytic diffuse lymphoma) with intermediate grade of malignancy. One case (19 years) died before treatment. The autopsy showed a large solitary tumor (18 cm in diameter) in the right liver lobe and two small nodules (1 cm) in the subcutaneous tissue of the abdominal wall. The other case (62 years), was treated by partial hepatectomy and chemotherapy. There was a round, solitary tumor (8 cm in diameter) in the left lobe. The patient remains without manifestations of disease relapse 18 months after the treatment. In both cases the tumor did not spread to lymph nodes, spleen and other viscera. PMID- 8240062 TI - [Upper gastrointestinal hemorrhage associated with chronic pancreatitis and pseudocyst: report of a case]. AB - The authors present a case of a 30 years old chronic alcoholic male with gastrointestinal bleeding. Celiac axis arteriogram demonstrated a false aneurysm of the anterior pancreatico-duodenal artery. Bleeding was controlled by embolization of the nourishing artery using the Gianturco-Wallace device and the patient left the hospital 2 days after the procedure. A partial revision of the pertaining literature is presented. PMID- 8240063 TI - [HLA and chronic alcoholic pancreatitis. A meta-analysis]. AB - The authors made a meta-analysis of nine important works about the possible relationship of IILA and chronic alcoholic pancreatitis. They concluded that there are actually three types more prevalent: A1, B13 and B18. PMID- 8240064 TI - Abnormalities of the interdigestive migrating motor complex in a patient with carcinoid syndrome. AB - Intraluminal pressures of the gastric antrum, duodenum and jejunum were recorded for 206 min after a 12 h fast in a patient with carcinoid syndrome due to neoplasia of enterochromaffin cells of the ileum and with hepatic metastases. The most conspicuous alteration was a tachyrrhythmia of 16 waves/min predominating in the activity fronts of both duodenum and jejunum. Periods of 11-12 waves/min frequency appeared irregularly and the simultaneous occurrence of frequencies of 11 waves/min and 16 waves/min was also recorded. The gastric antrum was fully quiescent throughout the study. The alterations observed are presumed to be produced by substances secreted by the carcinoid tumor. PMID- 8240065 TI - [Resection of colonic sessile polyp by strip biopsy]. AB - Sessile polyps of the colon can present difficulties when being resected by the usual method of polypectomy. Strip biopsy has been reported as effective to accomplish the resection of semipedunculated or sessile lesions. In this paper we confirmed the efficacy of strip biopsy to perform the resection of a sessile polyp of the colon. PMID- 8240066 TI - Delays by recalcitrant FDA, reluctant NIH, and fearful industry: the cost in human life, happiness, money, and loss of opportunity for American industry. PMID- 8240067 TI - Clinical experience and model analysis on beta-2-microglobulin kinetics in high flux hemodialysis. AB - Beta-2-microglobulin (beta 2M) is associated with amyloidosis. The study of beta 2M kinetics can provide information on the elimination of this uremic toxin. A beta 2M kinetic model modified from Gotch, considering the volume changes between intracellular, interstitial, and intravascular compartments and the generation stimulation and inhibition during hemodialysis is proposed. The clinical experiments on 8 stable hemodialysis patients treated with polysulfone (F80) and polymethyl methacrylate (PMMA, BK2.1p) 3 times a week were conducted. There was an 18% decrease of beta 2M clearance in the period from 30 to 180 min with a time averaged beta 2M clearance of 48 ml/min using polysulfone dialyzers (F80). In PMMA dialyzers, there was a 64% decrease of beta 2M clearance from 30 to 180 min with a time-averaged clearance of 56.3 ml/min. During hemodialysis, the generation rate was 0.379 mg/min in polysulfone and 0.828 mg/min in PMMA dialyzers. There was a stimulation generation of 0.309 mg/min in polysulfone and 0.749 mg/min in PMMA during hemodialysis. In conclusion, we provide a beta 2M kinetic model including volume changes, polymerization, generation stimulation, and inhibition that is similar to the human physiological condition. This model can be used for further clinical study. PMID- 8240068 TI - Serum melatonin and 6-sulfatoxymelatonin in end-stage renal disease: effect of hemodialysis. AB - The pineal hormone melatonin plays a role in the regulation of hypothalamic pituitary axis, target gland function, sleep pattern, mood changes, cellular immunity, antibody response, and skin pigmentation, nearly all of which are also affected by renal insufficiency. We measured serum melatonin and its main urinary metabolite, 6-sulfatoxymelatonin (6-SM), by radioimmunoassay in 11 patients with end-stage renal disease (ESRD) between the hours of 6 and 9 a.m. during hemodialysis and again on an off-dialysis day. The results were compared with those obtained in a group of normal control subjects. Predialysis serum melatonin and 6-SM levels obtained at 6 a.m. in the ESRD patients were comparable with those obtained in the normal control group. Serum melatonin concentration in the ESRD group fell approximately 25% during dialysis while 6-SM remained virtually unchanged. The changes observed on the off-dialysis day during the same time period were of nearly the same magnitude as those seen during dialysis (-32% and 1.4%, respectively). No significant difference was found in the concentration of either analyte in the blood entering and leaving the dialyzer. These observations suggest a lack of discernible removal of either compound by hemodialysis. In contrast to the ESRD patients, who showed a slow fall in serum melatonin and no significant change in serum 6-SM on both on- and off-dialysis days, the normal control subjects showed an expected reduction in serum melatonin (-43%) and a sharp fall in serum 6-SM (-53%) between 6 and 9 a.m.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240069 TI - The effect of interdialytic weight gain on predialysis blood pressure. AB - Interdialytic weight gain is believed to influence predialysis blood pressure. Since interdialytic weight gains vary among treatments for individual patients, blood pressure and weight gain data could be examined to determine how weight variations correlate with differences in blood pressure. Therefore, the quantitative effect on prehemodialysis blood pressure of typical interdialytic weight gains was prospectively studied in 19 nondiabetic patients on chronic hemodialysis. Over a mean of 23.6 treatments (range 17-25), the slope of each patient's prehemodialysis blood pressure versus excess weight (prehemodialysis weight minus baseline dry weight) was determined. The mean slope of the prehemodialysis mean blood pressure/excess weight regression line was 1.2 mm Hg/lb excess weight. No significant correlation was found between individual prehemodialysis blood pressure/excess weight slopes and patient age (r = 0.20), months on dialysis (r = 0.33), dry weight (r = 0.05), or mean excess weight (r = 0.19). Slopes did not differ for 3-day versus 2-day interdialytic intervals, hypertension-treated versus untreated patients, or men versus women. In 5 patients, individual prehemodialysis mean blood pressure/excess weight slopes were significantly greater than 0, averaging 2.4 mm Hg/lb excess weight (vs. 0.8 mm Hg/lb in the remaining patients). These 5 volume-responsive patients did not differ clinically from the 14 volume-resistant patients. The weight gains commonly observed in patients undergoing chronic hemodialysis have only a modest effect on prehemodialysis blood pressure in the majority of patients. PMID- 8240070 TI - Extracorporeal endotoxin removal by immobilized polyethylenimine. AB - The neutralization of bacterial endotoxins (ET) is still an unsolved problem in therapeutic medicine. The efficacy of anti-endotoxin antibodies or receptor antagonists and other substances interfering with the endotoxin-induced pathomechanisms is dependent on an intact cellular degradation system of the host. However, the phagocytosis function of that system seems to be impaired regularly in patients with intense or long-lasting endotoxemia or septic shock and in patients undergoing hemodialysis. Extracorporeal adsorption of ET might well be an effective support in the anti-ET therapy by lowering the amount of circulating ET and thus relieving the defense system of the body. In this work a new ET-adsorbent based on macroporous cellulosic beads with immobilized polyethylenimine (PEI) was tested for its ET-removal capacity in vitro. A test solution with 100 ng/ml ET from Escherichia coli 055:B5 was recirculated in a system containing the adsorbent beads. Polymyxin B immobilized to the same carrier was used for comparison. PEI as well as polymyxin B showed complete removal of ET from plasma and water as was measured by the Limulus Amebocyte Lysate (LAL) test (Chromogenix). The biocompatibility of the PEI absorber was superior to that of polymyxin B. The results indicate that the PEI absorber is of high efficacy and possibly of interest for the treatment of endotoxemia. PMID- 8240071 TI - Complement activation during tryptophan immunoadsorption treatment. AB - Antibodies against human lymphocyte antigens (HLA) are frequently seen among patients undergoing repeated renal transplantations. Graft survival can be improved by eliminating these antibodies by plasmapheresis before transplantation. In this study, we have tried a new extracorporeal procedure to remove the anti-HLA antibodies. An immunoadsorption column (IM-TR) with a matrix of polyvinyl alcohol (PVA) gel conjugated with a hydrophobic amino acid tryptophan was utilized. Previous results have shown that repeated IM-TR treatments are at least equally effective as plasmapheresis in reducing levels of specific immunoglobulins in treated patients. In this study, 7 HLA-immunized patients were treated before renal transplantation. Each patient was subjected to a total of 12 treatment sessions divided into 3 sessions per week. After each treatment session, the reduction of the immunoglobulins was less than what has been reported for plasmapheresis. This suggests that mechanisms other than immunoglobulin depletion are involved in the reduction of the total immunoglobulin levels. The IM-TR treatment resulted in a strong complement activation triggered by the alternative pathway. Since the adsorbed plasma was returned to the patient, exceedingly high levels of the activation fragment C3d (C3dg) were found in plasma during and after the treatment. We conclude that the extensive generation of C3dg may be one of the factors that plays a role in the reduction of the antibody levels since the C3dg fragment has been shown to down regulate the immune response. PMID- 8240072 TI - A new device for endothelial cell seeding of a small-caliber vascular prosthesis. AB - A device permitting homogeneous endothelial cell seeding of a small-caliber arterial prosthesis has been developed. The prosthesis is maintained firmly attached to a rotative scaffolding device. This device is activated by an electrical motor at constant and adjustable speed. The whole system is maintained at 37 degrees C in a cell culture incubator. The 4 mm internal diameter polytetrafluoroethylene (PTFE) prosthesis was coated with biological glue and seeded with human saphenous vein endothelial cells obtained by mechanical detachment. Cell seeding density was 2.10(4) cells/cm2 (Group A, n = 6) or 10(5) cells/cm2 (Group B, n = 6). Rotation speed was 8 revolutions per hour (rph) during 90 min. Analysis of the homogeneity of cell seeding was permitted by cell counts on five different segments of the prosthesis. Each longitudinal segment was analyzed at three different subsegments of the circumference. The average adhesion was 43 +/- 4% in Group A and 38% +/- in Group B of seeded cells. No difference could be observed between the different segments and subsegments. In the two groups, cells were spread, and in Group B, a complete endothelial cell layer was obtained on the graft surface. This study permits validation of the device to allow homogeneous cell seeding in an arterial prosthesis. PMID- 8240073 TI - Reoperations on prosthetic heart valves: an analysis of outcome. AB - To evaluate risks and complications of reoperations on prosthetic heart valves, we reviewed data on 70 patients who underwent reoperations because of prosthetic valve malfunction. Overall hospital mortality was 13% (9/70 patients). The common cause of death was low cardiac output syndrome following surgery (4 patients). Respiratory failure and mediastinal infection accounted for 2 deaths each, and neurological complication for 1 death. However, hospital mortality was different according to the risk factors; reoperations for prosthetic valve endocarditis (18%, p < 0.05), advanced New York Heart Association (NYHA) class (50%, p < 0.001), and emergency operation (33%, p < 0.005) were the significant risk factors. In contrast, advanced age, female sex, type of prosthesis, valve position, and diagnosis (leak, structural deterioration, or valve thrombosis) did not appear to be significant risk factors. There were 7 late deaths (4 valve related, 2 cardiac, and 1 noncardiac). Inasmuch as emergency operation, advanced NYHA class, and prosthetic valve endocarditis affected hospital mortality, these factors contributed to late death. Actuarial survival rate and freedom from valve related mortality at 10 years were 75.8 +/- 2.8% and 87.2 +/- 2.3%, respectively. There were 8 valve-related complications, and freedom from valve-related complications at 10 years was 73.5 +/- 3.5%. As judged by these data, hospital mortality and late survival can be improved if hemodynamic conditions leading to myocardial damage can be prevented. PMID- 8240074 TI - A fluid dynamic analysis of a rotary blood pump for design improvement. AB - The proper design of a left ventricular assist device (LVAD) requires an understanding of the pump's fluid dynamic and biocompatible properties. A hydraulically efficient system minimizes the power required for pumping. Biocompatibility refers to the ability to pump blood with minimal hemolysis and thrombus formation. Typically, shear stresses below a threshold level will not damage blood significantly. A fluid dynamic analysis of a prototype centrifugal pump designed for use as an LVAD was performed to establish flow characteristics. A flow visualization technique using Amberlite particles suspended in a glycerin/water blood analogue was used. The system was illuminated with a 1 mm planar beam strobed helium-neon laser, and the results were recorded photographically. An analysis of photographs revealed laminar and turbulent flows with vortices within an illuminated plane in both the inlet and outlet port areas. From these data, velocity and shear stress profiles were generated that showed possible areas of improvement. It was concluded that the outlet port design could be improved by changing its angle and the continuity of its expansion. The inlet port could also be improved by smoothing the transition area between the inlet tube and the pump body to allow for gradual acceleration of the entering fluid. PMID- 8240075 TI - Dialysis against a recycled albumin solution enables the removal of albumin-bound toxins. AB - The removal of protein-bound substances of pathogenetic relevance from blood is of therapeutic interest for drug intoxications, renal and liver failure, and metabolic disorders. Current methods using adsorbents are effective but often not specific enough. This work presents an alternative method that enables the dialyzability of albumin-bound toxins from plasma by the use of a high-flux dialyzer (F 60 Fresenius) and an albumin solution circulating on the dialysate side to increase selectively the affinity for albumin-bound toxins. This method resulted in effective removal of unconjugated bilirubin, drugs with a high protein-binding ratio (sulfobromophthalein, theophylline), and a protein-bound toxin (phenol). The additional removal of PBS could extend the applicability of dialysis, for example, to drug intoxications and liver failure or could improve the elimination of protein-bound uremic toxins in chronic renal failure. PMID- 8240076 TI - Urea kinetic modeling: comparing the options. AB - In this study 6 commercially produced kinetic modeling packages utilizing a variable volume, single pool urea model, as well as formulae to determine the delivery of therapy, have been compared by applying to each the same set of rigorously collected data for a group of 12 patients. Comparison of the kinetically derived parameters (urea generation rate [G], urea distribution volume [V], delivery of therapy [Kt/V], and normalized protein catabolic rate [nPCR]) showed that the values obtained for both G and V differed between packages owing to the numerical methods and the clearance used in the solution of the differential equations. Although a broad agreement between the values established for Kt/V and nPCR was noted, the 95% limits of agreement indicated that it would be prudent to exercise caution when comparing results established by different modeling packages. PMID- 8240077 TI - Dialysis treatment using an ethylene vinyl alcohol membrane and no anticoagulation for chronic uremic patients. AB - Anticoagulation used in hemodialysis treatment brings with it the risk of hemorrhagic complications and the possible consequences associated with chronic heparin administration. These problems have not been satisfactorily addressed to date. This study examined a new dialysis method that does not require the administration of anticoagulants. Dialysis is performed for 3 h with a continuous infusion of 750 ml of physiological saline solution in predilution mode and using filters made of ethylene vinyl alcohol copolymer membranes. Eleven patients with chronic uremia underwent more than 2,000 dialysis treatments performed with 9 episodes of coagulation of the dialyzer or blood tubings (0.43%). An evaluation of individual treatments revealed a high degree of biocompatibility and only a scanty activation of coagulation. Blood depuration efficacy was very good as evaluated from pretreatment and posttreatment routine blood chemistries. The technique described here represents a simple and effective method for performing regular dialysis treatment and does not require anticoagulant therapy. PMID- 8240078 TI - Catastrophic vasoconstriction after protamine reversal of heparin anticoagulation in the goat. AB - Catastrophic pulmonary vasoconstriction after protamine reversal of heparin anticoagulation was observed frequently in goats that underwent centrifugal biventricular bypass pump implant. Care should be taken when protamine is used to reverse heparin anticoagulation in goats. PMID- 8240079 TI - Dental arch and palate in Taiwan aboriginals--Ami, Bunun, Paiwan and Rukai tribes. AB - Differences in the dental arch and palate among Taiwan Aboriginals were investigated and compared with Taiwanese and Japanese. Plaster models were taken from 315 Taiwan Aboriginal adults, 185 Taiwanese and 426 Japanese between 1979 and 1989, and various dimensions were measured. There were no clear differences in the characteristics of the dental arch and palate between Taiwan Aboriginal tribes. However, they were found to differ from those of Taiwanese and Japanese. In a dendrogram constructed from a cluster analysis, two major clusters were found. Among males, the first cluster contained Paiwan and Rukai, the second contained Japanese, Taiwanese, Ami and Bunun. Among females, the first contained Japanese, Taiwanese, Rukai and Paiwan, the second contained Ami and Bunun. The Ami were close to Bunun and the Paiwan were close to Rukai among both males and females. These results indicate that Ami and Bunun, and Paiwan and Rukai, respectively, have close affinities. PMID- 8240080 TI - Effects of phosphoprotein on collagen fibril formation in vitro. AB - Transmission electron microscopy showed that when either dentinal phosphoprotein or calcium-treated phosphoprotein or phosvitin were introduced during type I collagen fibrillogenesis the fibrils formed were significantly wider and the cross-banding was more distinct than in the absence of phosphoprotein. The collagen fibril width also increased with increasing concentrations of these molecules. When either bovine serum albumin (BSA) or dephosphorylated dentine phosphoprotein were used, no differences in the fibril characteristics were seen when compared to the controls that contained no phosphoprotein or BSA. When these dialysed matrices were placed into mineralizing solutions, no mineral was observed in any of the samples. PMID- 8240081 TI - Morphometric variables of developing primary mandibular second molars. AB - Morphometric variables of second primary molar crowns from birth to completion were examined and measured. An image analysing technique comprising a photographic camera, a monitor, a computer with appropriate software and a digital caliper was used. Strong correlations were found between the 'external' aspect of the crown (mesiobuccal cusp height, mesiodistal and buccolingual dimensions and perimeter), and between all the 'occlusal' measurements of the crown, i.e. all the intercusp distances. However, poor correlations were found between the external and internal measurements of the crown such as the mesiobuccal cusp height and mesiobuccal-mesiolingual cusp-tip distance. It was also found that the mesiobuccal cusp serves as the apex of two triangles, the one formed by mesial cusps, and one formed by the distal cusps. The cusps on the mesial portion of the crown (ML, DB) are on a similar radius from the MB cusp, and the cusps on the distal portion are on a similar radius from the MB cusp. The small sample is a limitation but the results suggest a separate and independent pattern and rate of development of the external and the occlusal morphological variables of the tooth crown. Further research is needed on larger samples. PMID- 8240082 TI - The influence of basal anxiety on unstimulated parotid and submandibular saliva. AB - Stress modifies numerous physiological variables, such as salivary secretion. The relation between salivary constituents, biophysical variables and personality traits defined by the Cattell's anxiety scale was studied. The main result was, that in the absence of stress, no statistically significant differences are to be found between the three groups defined on the Cattell scale. PMID- 8240083 TI - Influence of age on the secretory rates of the human minor salivary glands and whole saliva. AB - This investigation comprised two studies of healthy, unmedicated individuals. The first measured the effect of collection time on the volume of secretions of the minor salivary glands at four specified areas of the lower lip of 13 individuals before and after a mild gustatory stimulus. The second (n = 51) monitored the influence of age and gender on the secretory rates of unstimulated labial, buccal and palatal salivary glands. Also, unstimulated and stimulated flow rates of whole saliva were monitored to provide a point of reference. Volumes of minor gland secretions were measured with a Periotron unit. Results of the first study indicated a linear increase in volumes with collection time (15, 30, 45 and 60 s). Flow rates were similar among the four labial sites, approx. 1 microliter/cm2/min, and were not influenced by mild citric acid stimulation. Results of the second study indicated that flow rates differed significantly (p = 0.0001) among the anatomical sites, with similar rates on the right- and left hand sides. Gender exerted no influence on flow from the minor salivary glands. Similarly, age exerted no influence on flow from the buccal or labial glands. However, the secretory rate for the palatal glands decreased significantly with age (r = -0.44; p < 0.005). As for unstimulated whole saliva, secretory rates were not influenced by age nor gender; rates for stimulated whole saliva increased with age (r = 0.31; p < 0.05). No association was detected between the flow rates of the whole saliva and that of the minor salivary glands. PMID- 8240084 TI - Stimulation of intradentinal dye penetration by feeding in the rat. AB - Dentinal fluid movement, as measured by intradentinal dye penetration (IDDP), may be under the control of an endocrine system that includes the parotid glands. It was earlier demonstrated that parotid hormone stimulates IDDP when infused into rats, and in the pig the onset of feeding signals the release into the circulation of immunoreactive parotid hormone (iPH), the titre of which remains elevated for more than 1 h after feeding. As, for technical reasons, it is impossible to measure iPH and IDDP activity in the same animal, the hypothesis that feeding causes IDDP stimulation in the rat was now tested. Feeding rats for 15 min stimulated IDDP to a level significantly higher than when fasting (0.418 +/- 0.040 versus 0.106 +/- 0.022, p < 0.001). Within the experimental conditions, IDDP stimulation lasted longer than 15 min. Sialoadenectomy before the feeding experiment did not change the feeding-stimulated IDDP response unless the parotid glands were removed. In parotidectomized and totally sialoadenectomized rats fed for 15 min, the level of IDDP was not different from that of sham-operated fasting animals. It is postulated that, in addition to the existence of possible local regulatory factors in the pulp, dentinal permeation may be under a physiological control mechanism involving a parotid endocrine function. It is also suggested that a hormonally controlled mechanism of dentinal fluid movement may play a systemic, protective role against the bacterial acidogenic challenge to teeth. PMID- 8240085 TI - Characterization of a system of mineralized-tissue formation by rat dental pulp cells in culture. AB - Pulp tissue was obtained from maxillary incisors of young adult male Wistar rats, minced and digested with 0.5% trypsin and 0.02% EGTA at 37 degrees C for 30 min. Dissociated cells were cultured with or without 10 nM dexamethasone using Eagle's minimal essential medium supplemented with 10% fetal bovine serum and 50 micrograms/ml ascorbic acid. Confluent cells were subcultured at 7 days and the medium further supplemented with beta-glycerophosphate (beta-GP). Dexamethasone in primary culture and/or secondary culture enhanced the formation of mineralized tissue while > 5 mM beta-GP was necessary for mineralization to occur. Biochemical analysis of the radiolabelled medium revealed that these cells produced type I, type I trimer and type III collagens. Analysis of [32PO4] labelled medium, using DEAE-Sephacel ion-exchange chromatography and sodium dodecylsulphate-polyacrylamide gel electrophoresis, showed that these cells produced phosphophoryn-like protein. These results indicate that some of the rat dental pulp cells in culture express an odontoblast-like phenotype. PMID- 8240086 TI - The effects of cytoskeletal inhibitors on the collagen gel contraction by dog periodontal ligament fibroblasts in vitro. AB - Periodontal ligament fibroblasts (PLF) were incorporated into small collagen gel discs (100 microliters, dia 10.0 mm), cultured in media containing various concentrations of cytochalasin D or colchicine and the diameters of the discs then measured. Cytochalasin D (3.1-800 nM) inhibited contraction of the gel discs in a dose-dependent manner and 800 nM of cytochalasin D completely inhibited contraction. This complete inhibition was still observed when cell density in the gel was increased. Although colchicine at a concentration of 0.98-15.7 nM also inhibited the contraction in a dose-dependent manner, contraction at 50% of the control level was still observed in the presence of a high concentration (0.25 mM, 100 micrograms/ml). These results strongly indicate that actin filaments are more intimately involved in PLF-mediated collagen gel contraction than microtubules. Although it is suggested that PLF are involved in tooth eruption, earlier studies have shown that microtubule inhibitors inhibit eruption of rat incisors whereas the microfilament inhibitor, cytochalasin B, has no effect. Thus, the PLF theory alone seems insufficient to explain the mechanism of tooth eruption. PMID- 8240087 TI - Molar odontogenesis in the trisomic 16 mouse. AB - Stocks used were male and female monozygotes for Robertsonian translocation specific for chromosomes 16 and 17 Rb(16.17)7Bnr and males and females homozygous for Robertsonian translocation for chromosomes 6 and 16 Rb(6.16)24Lub to produce double heterozygotes characterized as Rb(16.17)Bnr/Rb(6.16)24Lub. This study was based on 156 fetuses, of which 70 were normal (euploid/controls) and 86 were affected trisomics identified grossly by decreased size, shortened faces (flattened snouts), oedema, petechiae, open eyelids and dysplastic ears. Confirmation of trisomics included karyotyping metaphasic spreads. Throughout the five gestational days studied (14-18), trisomic fetuses exhibited developmental delays of up to 24 h. In general, tooth organs were smaller, hypocellular, hypoplastic and had a decreased blood supply. These differences were progressive and more pronounced in the later periods of odontogenesis, especially in the morpho- and histodifferentiation stages. PMID- 8240088 TI - Potassium currents in cells derived from human dental pulp. AB - Patch-clamp recording methods were used to monitor ion currents in tissue cultured cells derived from human dental pulp. Recordings were made in excised, outside-in or whole-cell patches. In single-channel experiments, the majority of patches contained a high-conductance (approx. 140-180 pS) K(+)-selective ion channel. The probability of the channel being in an open state was dependent on membrane potential, internal calcium and negative pressure applied to the cell membrane. Whole-cell recordings were consistent with these findings; in response to step-wise depolarizations of the cell membrane, most displayed a family of outwardly rectifying, barium-sensitive currents. In addition, a number of patches contained a second class of potassium channel of intermediate (approx. 85-100 pS) conductance, which was largely voltage insensitive and independent of calcium concentration. These results suggest that pulp cells contain a high-conductance potassium channel which probably underlies the outwardly rectifying current found at the whole-cell level. Further, the existence of mechanosensitive channels in these cells raises the possibility that the response to mechanical perturbation of dental pulp may be mediated, in part, by direct effects on odontogenic cells. PMID- 8240089 TI - The distribution and morphological characteristics of axons innervating the periodontal ligament of reimplanted teeth in cats. AB - Twelve weeks after reimplanting lower canines the distribution and characteristics of myelinated and non-myelinated axons within the periodontal ligament were investigated by light and electron microscopy. The ligament was examined at 2-mm intervals along the length of the root of four reimplanted teeth and of the contralateral canines, which served as controls. In each case the periodontal ligament was reinnervated. In two of the teeth, which showed extensive apical resorption, the innervation was much less than in control teeth. In two teeth showing minimal or no apical resorption the degree of innervation at the most apical level did not differ significantly from controls. PMID- 8240090 TI - Immunohistochemical mapping of epidermal growth-factor receptors in normal human oral soft tissue. AB - Epidermal growth-factor receptor (EGF-r) has been identified on basilar cells of stratified squamous epithelia and skin adnexa in man. Recent studies have mapped EGF-r to various oral cells in animals; however, complete mapping of EGF-r in normal human oral mucosa has not been done. Normal tissues from eight sites in human oral mucosa were examined for their expression of EGF-r using avidin-biotin peroxidase complex with mouse anti-EGF-r monoclonal antibody. Immunoreactivity was detected in palatal gingiva, buccal gingiva, soft palate, lateral tongue, dorsal tongue and floor of the mouth. The connective tissues of the periodontal ligament and dental pulp were non-reactive. EGF is known to exist in most body fluids, particularly saliva. In normal human mucosa, EGF is localized to connective tissue subjacent to epithelium. With the receptor in the overlying epithelium, a possible epithelial-mesenchymal interaction may exist between the receptor and ligand. A paracrine mode of action may be postulated, functioning to regulate the complex biological functions of the human oral tissues. PMID- 8240091 TI - Vitamin A supplementation for retinitis pigmentosa. PMID- 8240092 TI - A randomized trial of vitamin A and vitamin E supplementation for retinitis pigmentosa. PMID- 8240093 TI - A randomized trial of vitamin A and vitamin E supplementation for retinitis pigmentosa. PMID- 8240094 TI - A randomized trial of vitamin A and vitamin E supplementation for retinitis pigmentosa. PMID- 8240095 TI - A randomized trial of vitamin A and vitamin E supplementation for retinitis pigmentosa. PMID- 8240096 TI - Combined obstruction of the central retinal artery and vein associated with meningeal carcinomatosis. PMID- 8240097 TI - Endogenous endophthalmitis caused by Propionibacterium acnes. PMID- 8240098 TI - Preparing for managed competition. Utilization of ophthalmologic services varies by state. PMID- 8240099 TI - Ocular gene therapy. From fantasy to foreseeable reality. PMID- 8240100 TI - Clinical physiology of heritable photoreceptor diseases. PMID- 8240101 TI - Leber's hereditary optic neuropathy masquerading as tobacco-alcohol amblyopia. AB - OBJECTIVE: To determine the frequency of known primary mitochondrial DNA (mtDNA) mutations for Leber's hereditary optic neuropathy (LHON) in patients previously diagnosed as having tobacco-alcohol amblyopia. DESIGN: A case series of 12 patients with tobacco-alcohol amblyopia. Follow-up ranged from 2 months to 15 years. SETTING: Tertiary care. PATIENTS: Twelve patients diagnosed as having tobacco-alcohol amblyopia, based on the classic clinical presentation, were tested for all the known primary mtDNA mutations associated with LHON. All patients had a history of heavy alcohol or tobacco use or both. Twelve other patients who fit inclusion criteria were unable to be contacted or refused to participate in the study. MAIN OUTCOME MEASURES: Presence of a known primary mutation for LHON at nucleotide positions 11778, 3460, 15257, or 14484 of mtDNA. RESULTS: Two (17%) of 12 patients previously diagnosed as having tobacco-alcohol amblyopia tested positive for known LHON genetic mutations, one for the 11778 mutation and one for the 3460 mutation. CONCLUSIONS: The diagnosis of LHON should be considered in all patients diagnosed as having tobacco-alcohol amblyopia, particularly those with visual acuities of 20/200 or less. The availability of molecular genetic testing for LHON now allows confirmation of the diagnosis of LHON in patients who otherwise may be misdiagnosed. PMID- 8240102 TI - Heteroplasmy in Leber's hereditary optic neuropathy. AB - OBJECTIVES: To determine the incidence and clinical significance of peripheral blood heteroplasmy and the presence of normal and mutant mitochondrial DNA in Leber's hereditary optic neuropathy through evaluation of a large series of families with the 11778 mutation and to evaluate the pattern of transmission of heteroplasmy. DESIGN: We studied heteroplasmy in 75 visually symptomatic patients with the 11778 mutation and in 101 asymptomatic family members. We compared the incidence of heteroplasmy in these two groups, collected clinical information for each symptomatic patient, and calculated the incidence of heteroplasmy within each generation of the pedigrees. RESULTS: We detected heteroplasmy in 24 (14%) of the 176 persons tested. Kaplan-Meier life-table analysis suggests that heteroplasmic persons are more likely to remain asymptomatic than those who are homoplasmic mutant (males, P = .17; females, P = .14). However, heteroplasmic persons who become symptomatic do not seem to differ clinically from symptomatic patients who are homoplasmic mutant. Pedigree analysis reveals a strong tendency for progression from heteroplasmy toward homoplasmy in subsequent generations (P = .001). CONCLUSION: Heteroplasmy for the 11778 mutation seems to play a role in the clinical expression of Leber's hereditary optic neuropathy and tends to progress toward homoplasmy in successive generations. PMID- 8240103 TI - Identical twins who are discordant for Leber's hereditary optic neuropathy. AB - OBJECTIVE: Leber's hereditary optic neuropathy is a maternally inherited form of visual loss that is associated with several mitochondrial DNA mutations. These mitochondrial DNA mutations are not the sole determinants of visual loss, as epigenetic factors may play a pathogenetic role. To clarify the role of these factors, we studied two visually discordant twins and determined their zygosity and mitochondrial genotype. DESIGN: Case series. SETTING: Referral center. PATIENTS: Identical twin brothers from a family with the 11778 mitochondrial DNA mutation. MAIN OUTCOME MEASURES: Visual acuity, results of testing for visual fields (measured with static and dynamic perimetry) and color vision, and results of funduscopic examination; alcohol and tobacco use, head trauma, co-existent medical illness, and occupational exposure; and results of mitochondrial DNA analysis and determination of zygosity. RESULTS: The monozygous twin brothers have remained discordant for the development of optic neuropathy for 6 1/2 years despite harboring the identical homoplasmic 4216, 13708, and 11778 mitochondrial DNA mutations. CONCLUSIONS: The patients are visually discordant despite being genetically identical at the nuclear and mitochondrial levels. Epigenetic factors are important determinants of visual loss in Leber's hereditary optic neuropathy in these brothers. Among those factors studied in these patients, a substantial difference was noted in regard to occupational exposure to toxic substances. Epigenetic factors that may influence the clinical expression of the mitochondrial DNA mutations associated with Leber's hereditary optic neuropathy should be systematically studied. Risk-factor intervention strategies should be formulated and implemented. PMID- 8240104 TI - A maculopathy associated with the 15257 mitochondrial DNA mutation. AB - OBJECTIVE: To report a new retinal finding associated with the mitochondrial DNA mutation at nucleotide position 15257, a primary mutation associated with Leber's hereditary optic neuropathy. DESIGN AND PATIENTS: Clinical and historical data were collected for 24 visually symptomatic patients from 20 independent pedigrees with the 15257 mutation. RESULTS: Fundoscopic examination in three patients who presented with acute, bilateral visual loss revealed retinal pigment epithelial changes in the maculae accompanied by normal-appearing optic discs. The conditions of two of these patients were initially diagnosed as Stargardt's disease, and subsequent molecular genetic analysis revealed the presence of the 15257 mutation. The third patient underwent molecular genetic analysis several months after presenting with a presumed maculopathy. Two of the patients also demonstrated evidence of a concurrent optic neuropathy. CONCLUSIONS: The association of macular changes with Leber's hereditary optic neuropathy associated mitochondrial DNA mutation has not been previously reported. The mitochondrial DNA mutation at nucleotide position 15257 may cause a maculopathy as well as the typical optic neuropathy usually seen in Leber's hereditary optic neuropathy. A subset of patients whose conditions were diagnosed as Stargardt's disease may harbor a mitochondrial DNA mutation. These three cases illustrate the importance of molecular genetic testing in some atypical cases of optic neuropathies and maculopathies. PMID- 8240105 TI - Natural course of visual functions in the Bardet-Biedl syndrome. AB - OBJECTIVE: To determine the course of visual functions in patients with Bardet Biedl syndrome. PATIENTS AND METHODS: The 21 patients with Bardet-Biedl syndrome seen at Children's Hospital, Boston, Mass, had optotype and grating acuities and dark-adapted thresholds measured over time. Their ages at first visit ranged from 2 weeks to 23 years (median age, 6 years). The courses of the visual functions were analyzed with a random-effects model. RESULTS: Substantial declines in visual functions were found. On average, grating and optotype acuities declined 0.09 log units (roughly 1 line) per year, and thresholds increased about 0.19 log units per year. The rates at which these visual functions were lost and the predicted level of the visual functions at ages 11 to 12 years (the mean ages of measurement) varied among individuals. CONCLUSIONS: The visual prognosis for children with Bardet-Biedl syndrome is poor. The course of both central and peripheral visual functions is variable. PMID- 8240106 TI - Pseudo low penetrance in retinoblastoma. Fortuitous familial aggregation of sporadic cases caused by independently derived mutations in two large pedigrees. AB - OBJECTIVE: The disparate occurrence of few cases of retinoblastoma in the same extended pedigree confronts us with the unsolved problem of a low-penetrant autosomal-dominant trait vs fortuitous familial aggregation of sporadic cases. Determination as to whether the disease arises from a common inherited mutation or sporadic mutations has important implications for genetic counseling. This is illustrated in this report of two presumed low-penetrant retinoblastoma pedigrees characterized by two distantly affected relatives connected through apparently healthy carriers. DESIGN: We mathematically modeled the inheritance patterns and calculated the a priori relative probabilities of heredity with low penetrance vs chance occurrence of independent mutations for each pedigree. The derived odds clearly show that the disease, which occurred twice in each family, most likely resulted from unrelated mutations. To prove this, extensive DNA testing was conducted, including determination of intragenic RB1 DNA sequence polymorphisms and screening for mutation using the polymerase chain reaction coupled with single-strand conformation polymorphism analysis. PATIENTS: All living key members from both pedigrees were included. RESULTS: Consistent with our initial expectation, there was no common intragenic haplotype or common germ-line mutation that segregated with the disease phenotype in either of these two families. CONCLUSIONS: We therefore conclude that collateral incidence of retinoblastoma in these two pedigrees occurred by chance and not according to autosomal-dominant inheritance with low penetrance. Furthermore, our data provide the first evidence, to our knowledge, that related individuals may have independent mutations involving an identical gene locus, giving rise to an artefactual inheritance pattern. PMID- 8240107 TI - A new codon 15 rhodopsin gene mutation in autosomal dominant retinitis pigmentosa is associated with sectorial disease. AB - OBJECTIVE: To ascertain and characterize rhodopsin gene mutations in autosomal dominant retinitis pigmentosa and to correlate these mutations with the clinical phenotypes. METHODS: DNA was extracted from leukocytes, and the rhodopsin gene was amplified and analyzed using molecular-biological methods. Clinical and electrophysiological data were collected from patient charts. RESULTS: We found a disease-causing mutation that was previously undescribed, to our knowledge, for autosomal dominant retinitis pigmentosa within codon 15 of exon 1 of the rhodopsin gene. It was a single base-pair transversion (AAT to AGT) leading to a serine-for-asparagine substitution. This altered a glycosylation site in the intradiscal portion of the rhodopsin molecule. The pedigree examined demonstrated an inferior distribution of retinal pigmentary changes and predominantly superior visual field loss with relative preservation of electroretinographic amplitudes and good vision, which is consistent with sectorial or sectorial-like retinitis pigmentosa. CONCLUSIONS: A codon 15 rhodopsin gene mutation caused retinitis pigmentosa in the pedigree studied. There may be an association between intradiscal rhodopsin gene mutations and sectorial forms of retinitis pigmentosa. PMID- 8240108 TI - Dominant retinitis pigmentosa associated with two rhodopsin gene mutations. Leu 40-Arg and an insertion disrupting the 5'-splice junction of exon 5. AB - OBJECTIVE: To determine the phenotypes of two families in which retinitis pigmentosa cosegregates with a rhodopsin (RHO) gene mutation: a leucine-to arginine change at codon 40 (Leu-40-Arg) in one family, and a 150-base pair insertion that disrupts the RHO 5'-splice junction of exon 5 in another. PATIENTS: Three affected members of each family. RESULTS: The Leu-40-Arg mutation was associated with the onset of night blindness in the first decade of life. By the fourth decade, severe retinal functional loss was evident on dark-adapted static threshold perimetry, and electroretinographic responses were absent or barely detectable. In contrast, the RHO 150-base pair insertion was associated with the later onset of mild night vision difficulties; in two individuals, mild night vision difficulties were first noticed in the second decade while a third, a 25-year-old woman, was asymptomatic. Dark-adapted static threshold perimetry of this latter individual revealed a "regional" or class 2 pattern of retinal functional loss associated with equal loss of rod and cone electroretinographic responses. CONCLUSION: The RHO Leu-40-Arg mutation causes symptomatic retinal dysfunction by the end of the first decade while the insertion disrupting the 5' splice junction of RHO exon 5 causes later onset "regional" or class 2 retinal dysfunction. PMID- 8240109 TI - Variable retinal and neurologic manifestations in patients harboring the mitochondrial DNA 8993 mutation. AB - OBJECTIVE: Ophthalmologic and neurologic manifestations of the mitochondrial DNA mutation at position 8993 (MTATP*NARP8993) are reported and compared with previously published reports of patients with the 8993 mutation and other mitochondrial disorders. DESIGN: Pedigree analysis. SETTING: University referral center. PATIENTS: Eight subjects from two unrelated pedigrees that were positive for the mitochondrial DNA replacement mutation at nucleotide position 8993 were evaluated ophthalmologically and neurologically. RESULTS: Retinal abnormalities ranged from mild salt-and-pepper changes to severe retinitis pigmentosa-like changes with maculopathy. Neurologic manifestations were also highly variable and ranged from migraine headaches to severe dementia and Leigh's disease. CONCLUSIONS: The type and extent of retinal pigmentary changes and neurologic findings varied substantially, even among members of the same family. These changes, although not specific for the MTATP*NARP8993 mutation, are highly suggestive of mitochondrial disease. PMID- 8240110 TI - Phenotypic variation including retinitis pigmentosa, pattern dystrophy, and fundus flavimaculatus in a single family with a deletion of codon 153 or 154 of the peripherin/RDS gene. AB - BACKGROUND AND OBJECTIVES: Mutations of the peripherin/RDS gene have been reported in autosomal dominant retinitis pigmentosa, pattern macular dystrophy, and retinitis punctata albescens. We report herein the occurrence of three separate phenotypes within a single family with a novel 3-base pair deletion of codon 153 or 154 of the peripherin/RDS gene. DESIGN: Case reports with clinical features, fluorescein angiography, kinetic perimetry, electrophysiological studies, and molecular genetics. SETTING: University medical centers. PATIENTS: A 75-year-old woman, her two daughters (aged 44 and 50 years), and her 49-year-old son were screened for peripherin/RDS mutations because of the presence of multiple phenotypes within the same family. RESULTS: The mother presented at age 63 years with a profoundly abnormal electroretinogram (ERG) and adult-onset retinitis pigmentosa that progressed dramatically over 12 years, with marked loss of peripheral visual field. One daughter developed pattern macular dystrophy at age 31 years. At age 44 years, her ERG was moderately abnormal but her clinical disease was limited to the macula. Another daughter presented at age 42 years with macular degeneration and over 10 years developed the clinical picture of fundus flavimaculatus. Her peripheral visual field was preserved but her ERG was moderately abnormal. The son had onset of macular degeneration at age 44 years. Pericentral scotomas were present and the ERG was markedly abnormal. Fluorescein angiography revealed punctate pigment epithelial transmission defects. CONCLUSIONS: A 3-base pair deletion of codon 153 or 154 of the peripherin/RDS gene can produce clinically disparate phenotypes even within the same family. PMID- 8240111 TI - A new syndrome of myelinated nerve fibers, vitreoretinopathy, and skeletal malformations. AB - OBJECTIVE: To report a new vitreoretinopathy with high myopia, myelinated nerve fibers, and skeletal abnormalities. PATIENTS: A mother and daughter with a previously undescribed constellation of severe vitreoretinal degeneration with high myopia and extensive myelination of the retinal nerve fiber layer, and in addition, the daughter has limb reduction deformities. Vision was severely impaired in both patients and results of electroretinography indicated the presence of cone and rod dysfunction. CONCLUSION: This is a previously undescribed autosomal dominant vitreoretinopathy with congenital poor vision, vitreoretinal degeneration, myelinated nerve fibers, and skeletal deformities. PMID- 8240112 TI - Phenotypic variation in combined granular-lattice (Avellino) corneal dystrophy. AB - OBJECTIVE: To describe the phenotypic variation exhibited by members of families with combined granular-lattice (Avellino) corneal dystrophy. SETTING: We examined 40 patients (age range, 12 to 85 years) from six unrelated families with Avellino corneal dystrophy. This included the first individuals to ever be examined near the presumed site of origin in Italy. In addition, one family was the first to trace its origins to Germany rather than to Italy. We studied the phenotypic expression of the disease in the cornea, visual acuities, subjective complaints, complications, treatment, and histologic condition of these individuals. RESULTS: The granular stromal lesions reach their mature quantity and size early in life, and appear as either gray and crumb-shaped deposits or superficial with an annular and planar distribution. The lattice component appears gradually, beginning and maturing later in life. The phenotypic variation within families was found to be substantial. Widely variable proportions of lattice and granular changes were found within single sibships. Visual acuities ranged from 20/20 to 20/400. Recurrent corneal erosions were present but infrequent. Subjective complaints included glare and decreased night vision. Penetrating keratoplasty was required in one individual to restore vision. Histopathologic examination revealed typical amyloid and granular deposits. Granular deposits were found replacing Bowman's membrane and extending to the corneal surface. These deposits probably represent the cause of recurrent erosions. CONCLUSIONS: Combined granular and lattice corneal dystrophy may present with substantial phenotypic variation. The disease can be found in individuals who trace their ancestry to both Italy and Germany, a wider geographic distribution than previously proposed. PMID- 8240113 TI - Identification of a nonsense mutation at codon 128 of the Norrie's disease gene in a male infant. AB - OBJECTIVE: Norrie's disease (ND) is a rare X-linked hereditary disorder characterized by congenital blindness. A putative gene for ND has been isolated and mapped to Xp11.3. Four point mutations in this gene have been identified recently in patients with ND, thus providing strong evidence that this gene is associated with the disease. We report a new mutation. DESIGN: Clinical findings from the proband were correlated with results from DNA analysis. The proband's DNA was compared with that from his mother, an unaffected brother, and four unrelated normal males. PATIENT: The proband was a male infant referred for ocular evaluation at 3 months of age. INTERVENTIONS: The patient was evaluated with a general ocular examination at 4 months of age, a computed tomographic scan at 8 months of age, and then periodic follow-up examinations over the next 7 years. Blood samples were also collected from the proband, his family, and four unrelated normal males. DNA was extracted, amplified using polymerase chain reactions, and then cloned and sequenced. RESULTS/CONCLUSIONS: We identified a new mutation at codon 128 of the ND gene, a dinucleotide GC-to-AA substitution that changed the normal codon for cysteine, TGC, to TAA, which is a stop codon. Thus, this patient lacks the last six amino acids of the carboxyl terminus of the ND protein. The normal ND protein has 11 cysteines in conserved positions that are proposed to be functionally significant. The mutation at codon 128 occurs at the 10th cysteine and might be expected to alter the function of the ND protein. Since the phenotype of this patient is similar to those of other patients with point mutations in the ND gene, this mutation is likely to be the molecular basis of the phenotype. PMID- 8240114 TI - Negative-configuration electroretinogram in Oregon eye disease. Consistent phenotype in Xp21 deletion syndrome. AB - OBJECTIVE: To determine whether abnormal configurations on electroretinogram were a consistent finding in patients with Xp21 deletion and to characterize the associated ophthalmologic phenotype. DESIGN: Case series. SETTING: University hospitals and eye institutes. PATIENTS: Five patients with complex glycerol kinase deficiency (Duchenne-type or Becker's muscular dystrophy, glycerol kinase deficiency, and congenital adrenal hypoplasia) and demonstrated chromosomal deletions at Xp21. Control patients were matched by age. MAIN OUTCOME MEASURES: Clinical information was obtained from medical records. Complete ophthalmologic examinations were performed. Electroretinography was performed using a Ganzfeld technique and chloral hydrate sedation. RESULTS: We report the clinical features and abnormal configurations on electroretinograms of five patients with complex glycerol kinase deficiency, including follow-up studies on a previously described patient. The original patient had ocular hypopigmentation; four, strabismus; two, myopia; three, astigmatism; and one, symptomatic night blindness. All had negative configurations on scotopic electroretinograms showing a reduced amplitude B wave in the dark-adapted state. CONCLUSIONS: Our original report suggested a diagnosis of Aland Island eye disease, which appears to be an incomplete form of congenital stationary night blindness. Linkage data place Aland Island eye disease and congenital stationary night blindness at Xp11, whereas our patients had deletions at Xp21. The phenotype reported here may represent the effects of a single gene defect or the compound effects of the Xp21 contiguous gene syndrome (complex glycerol kinase deficiency). The phenotype is referred to as Oregon eye disease. PMID- 8240115 TI - Lifetime prevalence of ocular injuries from the Baltimore Eye Survey. AB - OBJECTIVE: To estimate the cumulative lifetime prevalence, nature, and consequences of ocular injuries in a multiracial urban environment. METHODS: Self reported lifetime ocular injuries were ascertained in a population-based sample of persons aged 40 years or older residing in 16 clusters in east Baltimore, Md. A total of 5308 subjects underwent a screening eye examination and were interviewed about lifetime ocular injuries. RESULTS: Eight hundred thirty ocular injury incidents were reported by 762 (14.4%) subjects. The maximum number of ocular injuries reported in a lifetime was four. At least one injury was reported by 22.5% of black men, 20.3% of white men, 12.2% of black women, and 7.7% of white women. No treatment was sought for 18% of these injuries (22% black and 14% white subjects). The prevalence of visual impairment due to trauma per 1000 people was 22.3 among black men, 7.4 among white men, 6.7 among black women, and 3.5 among white women. The prevalence of blindness due to trauma in at least one eye was 21.2 per 1000 among black men, 3.7 times the rate among white men. Based on reports of injuries in the year prior to interview, the annual incidence of medically treated ocular injuries per 1000 people was 5.6 among black men, 4.1 among white men, 5.3 among black women and 1.8 among white women aged 40 years and older. CONCLUSIONS: Lifetime prevalences of ocular injuries were similar among both black and white men, but the visual consequences of the injuries were more severe among black men. PMID- 8240116 TI - Hyaloid vessels of the human fetal eye. A scanning electron microscopic study of corrosion casts. AB - OBJECTIVE: Microscopic investigation of the hyaloid vascular system in 5-month old human fetuses. METHODS: Corrosion casting and light and scanning electron microscopy. RESULTS: The hyaloid artery ramifies into a tuft of vasa hyaloidea propria, which communicates with the posterior portion of the tunica vasculosa lentis, characterized by a network of anastomosing vessels. They further pass to the lateral portion of the tunica, acquiring a nonanastomosing palisadelike array and drain into the vessels of the ciliary processes or, after bending over the edge of the developing iris, drain into the outer choriocapillaris. The tunica vasculosa lentis vessels also communicate with the pupillary membrane, a system of vascular arcades arranged in several interconnected tiers, supplied by the terminal branches of the long posterior ciliary arteries. In tunica vasculosa lentis, arterioles seem to pass directly into veins, without forming a capillary bed. CONCLUSIONS: At the investigated developmental stage, the fully developed hyaloid system enters its subsequent involution, and the vessels nourishing the vitreous have already involuted. The system is generally similar to that observed in other mammals. PMID- 8240117 TI - Orbital fat prolapse. PMID- 8240118 TI - Under-reporting of psychiatric illness. PMID- 8240119 TI - Non insulin dependent diabetes mellitus. Rational approach to management. AB - This article emphasises the importance of the underlying insulin resistance in NIDDM and its relationship to other associated disorders. The need for screening individuals at risk, with the aim of preventing micro and macrovascular complications, is discussed. Major lifestyle changes are advocated: weight loss, exercise, cessation of smoking and reduction of alcohol intake, dietary changes and self foot care. Patient education and self monitoring of blood glucose play a part in involving the patient in his or her own management. There is much benefit to the patient from the monitoring and early treatment of blood pressure, lipid levels and any diabetic complications that develop. PMID- 8240120 TI - Diabetes and vascular disease. AB - Reduction of morbidity and mortality rates from cardiovascular disease requires a dual approach: public health measures for the whole community and individual assessment and treatment for those people at high risk. The presence of diabetes identifies patients who require particular attention for risk factor assessment and prevention of macrovascular disease. PMID- 8240121 TI - Which insulin, when? PMID- 8240122 TI - The impact of diabetes on the lower limb. Give diabetes an inch and it will take a foot. AB - The author describes the role of vascular disease, neuropathy, trauma, and impaired inflammatory response and wound healing in the pathogenesis of foot disease. He discusses the general principles of management. Important management issues are highlighted including the need to look at the feet, to teach the diabetic how to care for them and how to get appropriate advice. PMID- 8240123 TI - Diet and Type 2 diabetes. Guidelines for general practice. AB - Type 2 diabetes (NIDDM) is common and increasing and results in enormous costs to all concerned. The average Australian diet is inappropriate for this condition, which regular exercise and healthy eating could delay or prevent even in high risk groups (family history, age over 50 years, over-weight). This article outlines a practical programme that general practitioners can use for people with, and people at risk of, Type 2 Diabetes and provides a comprehensive Further reading list. PMID- 8240124 TI - Diabetes shared care: a model. AB - Increases in both the cost of providing quality diabetes care and in the number of people developing diabetes have led to a search for more cost effective models of care. This paper outlines a shared care scheme that has benefits for the patient, the outpatient clinic and the general practitioner. PMID- 8240125 TI - Audit of a general practice. AB - A clinical activities audit is a method of obtaining interesting information about one's general practice. It is likely to generate improvement in practice, thereby fulfilling the most important requirement of quality assurance, even in the absence of nationally accepted standards of general practice. A five-day audit of clinical activities in one practice is detailed, and some of the changes that this prompted in consultation time management, investigation and referral rates, and conditions managed are frankly discussed. PMID- 8240126 TI - Neck sprain. AB - Neck sprain is characterised by 'soft tissue' lesions in the cervical discs, synovial joints, muscles and sometimes the nerve roots. Peripheral nociception in the outer annulus, joint capsules, muscles, meninges, arteries and nerves can cause neck pain, with headache, or upper torso and upper limb pain. Careful physical examination, or accurately placed local anaesthetic injections, may localise the sources of pain. PMID- 8240127 TI - Procedural medicine. Is your number up? AB - It is commonly argued that maintaining competence in procedural medicine requires the continuing practice of minimum numbers of procedures. While these continuing practice requirements may pose few problems for doctors working in city or regional hospitals, they may be impossible to meet in rural general practice. The authors review the literature relating to psychomotor skills in procedural medicine and argue that there is little or no evidence to justify the judging of competency by numbers of procedures performed. Instead it appears that with appropriate initial training, doctors can be expected to retain their psychomotor skills even after lengthy periods without practice. PMID- 8240128 TI - Will facilitators be acceptable in Australian general practice? A before and after feasibility study. AB - AIM: To describe the impact of the 'facilitator model' of promotion on two Australian general practices. METHOD: Attitudes and health promotion levels were evaluated before and after the facilitator worked with two practices. RESULTS: Simple changes to patient records were the most successful. CONCLUSION: Facilitators are acceptable and long term organisational changes can be achieved. PMID- 8240129 TI - Louis Ariotti, MBE, OAM. A GP-surgeon in the mulga. PMID- 8240130 TI - Mrs Cleary's pain relief. AB - This case illustrates some common problems faced in general practice--the effects of personality and family dynamics on symptom reporting, the balance between over investigation and under-investigation, and the dilemma of when to refuse to comply with patient requests for medication. Dealing with these is the art of good medicine. PMID- 8240131 TI - The dos and don'ts of prudent practice. PMID- 8240132 TI - A tale from country practice. Poet's day. PMID- 8240133 TI - Funding for research. AB - With the exception of pharmaceutical companies, all the funding organisations have a standard application and information form outlining the areas of research they wish to encourage and fund. Many of the organisations advertise nationally, indicating closing dates and contact numbers for further information. It is important to contact fund administrations to gain feedback on the direction of your project while it is still being drafted. This article has nominated just a few of the funding sources available to general practice related researchers. It is simply a matter of becoming familiar with the variety of sources and the types of projects that they fund. By applying only to common knowledge sources such as GPEP, researchers are ignoring the availability of alternate funding sources that may be even more appropriate to their project. PMID- 8240134 TI - Initial management of burns to the hands. PMID- 8240135 TI - Health Communication Network. PMID- 8240136 TI - Thyroxine dosage. PMID- 8240137 TI - General practice standards. PMID- 8240138 TI - A novel approach to the design of a Wilson referenced ECG amplifier. AB - The Wilson Central Terminal consists of three limb electrodes connected through a resistor network at the inverting input to the ECG amplifier. It is often used as a zero potential reference in ECG recordings. In this paper, the implications of using a Wilson central reference on the overall amplifier's common-mode rejection ratio and noise specifications is analysed. It is shown that the Wilson reference can degrade the overall amplifier specifications. The design of a Wilson referenced amplifier is then described which shows this to be true. A novel approach to the design of a Wilson referenced ECG amplifier is then presented, whereby the reference network is moved from the input to an intermediate stage of the amplifier. An analysis on the improvements achieved over the conventional approach is given. An amplifier design using the new approach is then described. Tests results showed a significant improvement in amplifier common mode rejection ratio and noise specifications when compared to the conventional design. PMID- 8240139 TI - Bone lead measurement using X-ray fluorescence. AB - An in-vivo x-ray fluorescence (XRF) bone lead analyser, based on the Somervaille design, has been constructed and tested as the first step towards developing a deep bone lead analyser to expand the range of bone tissue available for assessment, to include more deep seated bones such as the femur & vertebrae. These bones represent a major component of the total body bone tissue and therefore the lead body burden. A new test analyser, constructed by modification to the basic design of a superficial bone analyser, has been used to investigate the feasibility of using changes in source-detector geometry and collimation to improve deep bone lead sensitivity for in-vivo measurement techniques. Initial results indicate that the relative detection sensitivity of the test analyser is approximately 6 times that of the Somervaille based system. The results of examination of a series of lead loaded plaster of paris phantoms (0 ppm - 360 ppm) confirmed there was good correlation (r = 0.996, p < 0.0005) between the measured ratio (lead X-rays/coherent scatter photons) and the lead concentrations using data from both XRF analyser systems. PMID- 8240140 TI - A Monte Carlo technique to establish the water/tissue equivalence of phantom materials. AB - The quality of phantom materials is crucial for accurate dosimetry in radiotherapy. A wide range of factors such as density, electron density and elemental composition can influence the radiation properties, and hence the absorbed dose, of materials. New materials can be tested by direct measurements which requires considerable time and the availability of relatively large amounts of the material. Alternatively, the dosimetric properties of a proposed phantom material can be compared to those of water or tissue using Monte Carlo calculations. The aim of this study was to evaluate the use of a Monte Carlo technique for the investigation of the water/tissue equivalence of phantom materials. The material used for this investigation was Standard Dosimetry Agarose (SDA) gel, which is useful in MRI dosimetry. Depth doses in gel and water were calculated for mono-energetic electron beams of 6, 12 and 20 MeV, and photon beams of 60 keV and 6 MV. For each radiation quality the depth dose distributions are in close agreement. PMID- 8240141 TI - Development of a paediatric indirect calorimeter. AB - An indirect calorimeter was developed to measure resting energy expenditure (REE) and respiratory quotient (RQ) in a clinical paediatric population. The design is suitable for any child spontaneously breathing room air. Measurements are possible from small babies to large hypermetabolic adolescents due to the ability to set and control flow from the patient headbox across the range 2-40 l/min. The calorimeter is mobile, it is electrically safe and the patient interface may be disinfected. An alcohol burning technique, that can emulate a clinical test for children of various sizes, is used to check the system, which has a high level of accuracy across a range of conditions. The indirect calorimeter has been successfully used to measure a large number of children of different sizes. PMID- 8240142 TI - Readout of thermoluminescence dosimetry chips using a contact planchet heater. AB - Thermoluminescence dosimetry (TLD) is performed in many radiotherapy departments using LiF chips which are evaluated by means of a contact heater. The aim of the present study was to investigate the influence of the TL chip position on the heating planchet and to study the self absorption of light in the chips during readout. Experiments were performed using standard LiF chips (3.1 x 3.1 x 0.89 mm3) on a circular heating planchet with a diameter of 13 mm. A two step readout cycle (10s 160 degrees C, 10s 300 degrees C) was used and the integral under the luminescence curve during the second step evaluated. Moving the chip away from the centre of the planchet without adjusting the temperature cycle leads to a peak broadening which can result in a signal loss of up to 40%. The use of a purpose build positioning device tended to improve the reproducibility of the readings. Self absorption of light in a normal chip on the heating planchet was studied by exposing chips in the strong dose gradient of the build up region of a 18 MV X-ray beam. The dose throughout the chip was investigated using a stack of 0.14 mm thick chips. In a 5 x 5 cm2 field the dose varied from 7% at the surface facing the beam to 23% at the opposite side of the chip. Reading the chips with the side facing the beam up in the reader resulted in a 1.9% smaller dose reading than with the chips upside down.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240143 TI - Performance characteristics of a laser based planar imaging system. AB - With the advent of lasers and high sensitivity detectors, there has been a recent renewed interest in medical imaging at visible wavelengths. This paper reports on the investigation of the spatial resolution of a prototype laser based planar imaging system by examining the transmission of the laser beam through a phantom medium having similar absorption and scattering properties to that of soft tissue. The results of this work, whilst supporting the conclusions of other recent publications, have demonstrated a previously unreported phenomenon. With the experimental configuration described in this paper, opaque objects, in the form of thin wires, are equally well resolved at similar distances from both the input and exit boundaries of the scattering medium. The spatial resolution is a maximum nearest these boundaries. PMID- 8240144 TI - The costs of costing. PMID- 8240145 TI - Radiofrequency catheter ablation for supraventricular tachycardias. PMID- 8240146 TI - Intravenous aminophylline confers no benefit in acute asthma treated with intravenous steroids and inhaled bronchodilators. AB - BACKGROUND: The role of intravenous aminophylline in acute asthma is unclear despite meta-analysis of many studies comparing aminophylline with other bronchodilator therapies. AIMS: The aim of this study is to determine whether continuous aminophylline infusion confers any benefit in acute severe asthmatics treated with intravenous steroids and inhaled bronchodilators. METHODS: The study was randomised, double-blind and placebo-controlled. All patients received nebulised salbutamol (1 mL of 0.5%) and ipratropium bromide (1 mL of 0.025%) with glycol diluent (1 mL) at 0, two, four, six, eight and 12 hours, and six-hourly thereafter. In addition all patients were given intravenous hydrocortisone 250 mg six-hourly and oxygen to maintain normoxia. Aminophylline infusions were adjusted to maintain therapeutic levels. Peak expiratory flow rate (PEFR) was measured before and after nebulised bronchodilator on a two-hourly basis in the Emergency Department (ED) and six-hourly on the inpatient wards. RESULTS: Thirty-one patients were clinically sufficiently improved within 12 hours to be discharged home from the ED. The remaining 28 patients were admitted to the inpatient ward for a total trial duration of 48 hours. No significant difference was found between the placebo and treatment groups for measurements of PEFR, or for the duration of stay of the patients in hospital. The power of the study was 80% for a 25% to 33% difference at a 5% level of significance. Presentation values of PEFR and arterial blood gases did not predict which patients would require inpatient admission and which could be safely discharged home from the ED. PMID- 8240147 TI - Retention and survival of hostel residents--a 12 year study. AB - BACKGROUND: The admission of a proportion of disabled people to hostels is inevitably followed by their transfer to nursing homes. Our hypothesis was that such admissions are justified in terms of quality of life and the cost to the community, notwithstanding the necessity of subsequent transfer. AIMS: To test this hypothesis by measuring the retention and survival times of residents in hostel and in nursing home; to consider the relevance of these factors to the future policy of the two institutions. METHODS: A retrospective study was made of 159 residents admitted over a period of 12 years to a hostel with 32 places. Times spent in the hostel and in the nursing home were recorded. Probabilities of survival in hostel and in nursing home were calculated according to the Kaplan Meier method. Comparison with the expected survival of a matched cohort of the total population was determined. Estimation was made, using the SAS software package, of the likely number of places needed in nursing homes for residents following transfer. RESULTS: Although the majority of hostel residents eventually needed nursing home care, a worthwhile proportion of their total institutional time (approximately two-thirds) was spent in the hostel. Ongoing support from the personnel in a geriatric service is likely to increase retention time in the hostel. Because of the ultimate outcome for the majority of residents, planning for hostel care should include consideration of places needed in nursing homes. PMID- 8240148 TI - Visualisation of arterial structure in vivo with intravascular ultrasound. AB - BACKGROUND: Contrast angiography provides a silhouette of the arterial lumen, but does not give information about arterial wall structure. Catheter-tip ultrasound transducers can now provide a cross-sectional image of the arterial wall. This study examined the pathological correlation of intravascular ultrasound images and the accuracy of ultrasound measurements of vascular geometry. METHODS: Intravascular ultrasound images were obtained with a mechanically rotated catheter-tip transducer and recorded on videotape. Initial validation studies were performed in fresh, post-mortem arterial specimens, which were filled with saline at physiological pressures. Ultrasound images at specific sites were compared with the pathological findings at that site and measurements of luminal diameter were compared with corresponding angiographic measurements. Subsequently, intravascular ultrasound was employed to examine the aorta, ilio femoral and coronary arteries in patients undergoing balloon angioplasty. RESULTS: The pathological correlations showed that intravascular ultrasound can detect early initial thickening and mild atherosclerotic lesions that do not result in luminal deformation. Ultrasound images provided definition of calcified, fibrotic and lipid-filled lesions. Ultrasound measurements of luminal diameter correlated well with pathology measurements (r = 0.93), as did ultrasound measurements of plaque area (r = 0.89). The in vivo studies demonstrated that intravascular ultrasound can define atheroma lesions not evident on contrast angiography and permits detailed evaluation of the results of interventions such as balloon angioplasty. CONCLUSIONS: Intravascular ultrasound provides a unique window upon arterial structure and pathology in humans. Ultrasound images allow accurate measurements of vascular geometry and define early atheromatous lesions that are not evident with angiography. PMID- 8240149 TI - The effect of pregnancy on the epilepsies: a study of 37 pregnancies. AB - BACKGROUND: Although studies have assessed the effect of pregnancy on epilepsy, usually the types of epilepsy are not differentiated and most have not included a control group, despite the natural history of epilepsy including fluctuations in seizure frequency. AIMS: To assess the effect of pregnancy on seizure frequency and compare this with changes in seizure frequency in non-pregnant patients. In addition, the relationship between seizure frequency during pregnancy and epilepsy type, seizure frequency prior to pregnancy and duration of epilepsy will be assessed. METHODS: Seizure frequency was assessed retrospectively in 37 pregnancies from 24 women by comparing the seizure number for the nine-month period prior to pregnancy with the number during the pregnancy. An increase in frequency was defined as a 50% or greater increase in the number of seizures. Twenty-four non-pregnant women, matched for age and epilepsy type, were included to assess fluctuations in control. RESULTS: In 41% of pregnant women, there was an increase in seizure frequency, in 51% no change and in 8% improvement. In the control group, 24% had an increase, 65% no change and 11% improvement. There was no correlation between seizure frequency during pregnancy and epilepsy type and seizure frequency prior to pregnancy, but those with longer duration of epilepsy were more likely to deteriorate (p < 0.05). Alterations in anticonvulsants to reduce the risk of teratogenicity was a common identifiable cause of deterioration in control. CONCLUSIONS: Significant random fluctuations in epileptic control occur, but pregnancy may have a deleterious effect on epilepsy, particularly when appropriate therapy is withdrawn to reduce teratogenicity. PMID- 8240150 TI - Percutaneous transhepatic measurement of the pressure gradient between the portal and hepatic veins. AB - BACKGROUND: Knowledge of the portal pressure may be of value in the assessment of patients with chronic liver disease but its measurement is problematic. AIMS: To evaluate the ease and safety of percutaneous transhepatic measurement of the pressure gradient between the portal and hepatic veins and to determine directly the need for an internal zero. METHODS: Sixty-one patients undergoing liver biopsy for suspected liver disease had pressures in branches of portal and hepatic veins measured using a flexible 22G (Chiba) needle. RESULTS: The procedure was successful in all patients, took less than ten minutes in most, and was associated with minimal discomfort. Post-procedure morbidity was similar to that of liver biopsy. Portal pressure using an external zero (either puncture site or sternal angle) was inaccurate compared with pressures obtained using the generally accepted gold standard internal zero, hepatic venous pressure, and led to incorrect classification of the presence or absence of portal hypertension in at least 10% of patients. Variations in hepatic venous pressure were not predictable on clinical grounds. The only histopathological feature predictive of portal hypertension was cirrhosis, 20 of 25 patients with and four of 36 patients without cirrhosis having portal hypertension. Of routine biochemical and haematological tests, only plasma albumin and platelet count jointly (and negatively) predicted hepatic venous pressure gradient on multiple regression analysis (R2 = 0.40). CONCLUSIONS: The use of an internal zero is essential for accurate measurement of portal pressure and this can be achieved safely using the percutaneous, transhepatic route in patients with well compensated liver disease. PMID- 8240151 TI - Treating hypercholesterolaemia with HMG CoA reductase inhibitors: a direct comparison of simvastatin and pravastatin. AB - BACKGROUND: Simvastatin and pravastatin are both competitive inhibitors of the rate limiting enzyme for cholesterol biosynthesis (HMG CoA) reductase, but data from individual clinical trials suggest significant differences in potency for cholesterol reduction between the two drugs. AIM: To assess any differences in efficacy and safety between simvastatin and pravastatin in a direct, comparative study. METHODS: A double-blind, double-dummy, randomised study design was used, involving 48 patients with primary hypercholesterolaemia. Following a 6 week placebo baseline period, patients were randomly allocated to treatment with either simvastatin or pravastatin, commencing at a dose of 10 mg daily. The dose levels were titrated up to the recommended maximum effective dose of 40 mg daily at 6 weekly intervals if LDL cholesterol levels remained > or = 3.4 mmol/L. After 18 weeks of therapy, all patients were transferred to simvastatin therapy for a further 6 weeks, continuing at their week 18 dose level. Patients complied with a standard lipid lowering diet (containing < 30% of energy as total fat) throughout the study period. RESULTS: Over the 18 week direct comparison of the two drugs, there was a significant difference (p < 0.001) in response between simvastatin and pravastatin for reduction in levels of total cholesterol (32% vs 21% respectively), LDL cholesterol (38% vs 27%) and apolipoprotein B levels (34% vs 23%). No significant difference in drug effect was seen for the small reduction in levels of apolipoprotein AI (5% vs 6% respectively), nor for the increased levels of apolipoprotein AII (14% vs 11%) and HDL cholesterol (11% vs 7%). Lp(a) levels remained unchanged. When pravastatin was replaced with simvastatin for the final 6 weeks of the study in the 23 patients initially randomised to pravastatin, there were further reductions (p < 0.01) in total and LDL cholesterol, and apolipoprotein B. These results establish the advantage of simvastatin over pravastatin in terms of efficacy, for the treatment of primary hypercholesterolaemia. PMID- 8240152 TI - Chemotherapy made easier. PMID- 8240153 TI - Recombinant antibodies: back to the future. PMID- 8240154 TI - Heart transplantation in Australia--the effect of distance from the transplant unit. PMID- 8240155 TI - Vertebral osteomyelitis in advanced breast cancer. PMID- 8240156 TI - Delayed reactivation of visceral leishmaniasis complicating HIV infection. PMID- 8240157 TI - Cyclophosphamide related hepatotoxicity. PMID- 8240158 TI - House dust mite increase in Wagga Wagga houses. PMID- 8240159 TI - Adrenal and testicular tumours and azoospermia in congenital adrenal hyperplasia. PMID- 8240160 TI - Concurrent hepatitis B and C infection treated successfully with alpha interferon. PMID- 8240161 TI - Melioidosis of the brain presenting with a stroke syndrome. PMID- 8240162 TI - Cryoglobulinaemia, hepatitis C and interferon therapy. PMID- 8240163 TI - Ethics and clinical medicine. PMID- 8240164 TI - Ross River virus disease--the first recorded outbreak? PMID- 8240165 TI - Should doctors be concerned with efficiency? PMID- 8240166 TI - Radiofrequency catheter ablation of tachyarrhythmias--adult and paediatric experience. AB - BACKGROUND AND AIMS: Until recently, curative therapy for supraventricular tachycardia (SVT) was possible only with map-guided cardiac surgery. The ability to ablate permanently the reentrant circuit responsible for tachycardia using radiofrequency electrical current (RF) delivered through a catheter has now allowed cure of SVT by non-surgical means. We present our initial experience with this technique in both adult and paediatric patients. METHODS: In 53 patients of median age 32 years, 56 radiofrequency catheter ablation procedures were performed for control of tachyarrhythmias. Fourteen patients were aged 18 years or less. Indications for RF ablation were SVT due to an accessory pathway (n = 27), SVT due to atrioventricular junctional reentry (AVJRT) (n = 22), incessant SVT (n = 2), ventricular tachycardia (n = 2), and uncontrolled atrial fibrillation (n = 3). General anaesthesia was used in 88% of procedures. Ablation was preceded by electrophysiological testing to confirm tachycardia mechanism in all cases. RESULTS: The ablation procedure was completely successful in 49/56 cases (87.5%) and partially successful in another two cases. Only five procedures (9%) failed. Median procedure time was two hours, median fluoroscopy time was 24 min, and the median number of RF applications was four. There were no major differences in results comparing adult and paediatric patients. A transient ulnar nerve palsy was the only major complication observed. After a follow-up of one to 15 months, tachycardia has recurred in two patients (4%). Of the total 53 patients, 90.5% are currently asymptomatic and off all cardiac drug therapy. RF catheter ablation appears to be an effective, curative, and relatively safe alternative to long term drug therapy in the management of troublesome SVT. PMID- 8240167 TI - Cost effectiveness of radiofrequency catheter ablation in the treatment of symptomatic supraventricular tachyarrhythmias. AB - BACKGROUND: The recent advent of radiofrequency (RF) catheter ablation as a curative therapy for supraventricular tachyarrhythmias has challenged the role of long term drug treatment, which is essentially a palliative therapy. To date, however, no data have been published on the cost-effectiveness of RF ablation as compared with drug treatment in the Australian setting. AIMS: The study aimed to compare actual and projected costs of these two treatment options in a consecutive group of patients having RF ablation as treatment for symptomatic tachyarrhythmias. METHODS: The cost effectiveness of RF catheter ablation was assessed in 26 patients having RF ablation, using a hypothetical model of continued drug therapy in the same group of patients. A 'cost saving' criterion was used for cost effectiveness. Actual costs for the RF ablation and for continued drug therapy were based on data from medical records and from the answers to a detailed patient questionnaire. Analysis included costs of prior diagnostic electrophysiology (EP) study (17/26 patients), general anaesthesia (GA: 20/26 patients), post-ablation echocardiography (10/26 patients), and late follow-up EP study (7/26 patients). The in-hospital stay for the RF ablation was two days in all cases, and no patient required implantation of a permanent pacemaker. The RF ablation procedure was successful in 23/26 patients (88.5%) with late recurrence of tachycardia in one patient. After a median follow-up of nine months, 22/26 patients no longer require antiarrhythmic drug therapy. RESULTS: The mean per patient cost of RF ablation was $4067 in the study group. This reduces to $2546 if prior EP study and GA are excluded. The mean per patient cost of continued medical therapy was $700 per year. Extrapolating over 20 years and allowing for an annual 5% inflation factor, RF ablation becomes cost saving in 5.5 years (3.8 years if prior EP and GA are excluded). Over 20 years, continued drug therapy would be four to five times more expensive than RF ablation in the patient study group. We consider RF ablation to be a cost effective alternative to long term drug therapy in patients with supraventricular tachyarrhythmias. PMID- 8240168 TI - Government animal health laboratory services in Western Australia. PMID- 8240169 TI - Kanamycin concentrations in synovial fluid after intramuscular administration in the horse. AB - Six adult ponies were injected in the same intramuscular site with kanamycin sulphate (10 mg/kg). Two hours later, arthrocenteses of the right metacarpophalangeal, radio-carpal, intercarpal, tibio-tarsal and metatarsophalangeal joints were performed within 3 minutes. Arthrocenteses of the same joints on the left side were conducted 5 hours later. When expressed as a percentage of plasma drug concentration, differences in synovial fluid drug concentration between the joints sampled at 2 and 5 hours after injection were not detected. PMID- 8240170 TI - Distribution of lesions in ovine salmonellosis. AB - Four forms of salmonellosis were recognised in feedlots and during transport by sea: septicaemic, and acute, subacute and chronic enteric. The severity and distribution of lesions in the enteric forms varied according to the progression of the pathological process. The acute disease involved the abomasum and small intestine whereas the subacute disease centred on the lower small intestine and upper large intestine. Chronic disease involved considerable mucosal repair in the ileum, caecum and proximal colon. Septicaemic salmonellosis was often accompanied by acute enteritis and occasionally by cholecystitis. S typhimurium was the most frequently encountered serotype. PMID- 8240171 TI - Epidemiology of salmonellosis in the live sheep export industry. AB - Salmonellosis in sheep was studied on 7 commercial voyages from Australia to the Middle East and in one animal house study. Faecal excretion of salmonellas was not a good indicator of pathological changes in the gastrointestinal tract. The ratio of the proportion of sheep excreting salmonellas to the proportion with histological lesions of salmonellosis ranged from 1:1 to 23:1. Increasing excretion rates in our studies indicated that most sheep would be exposed to infection, although deaths from salmonellosis were not spatially or temporally clustered as would be expected if challenge alone was a sufficient cause. We considered that inappetence predisposed to death from salmonellosis, and that sheep with inappetence were likely to be randomly distributed aboard ship. In the animal house study, histological lesions of salmonellosis were exclusively in inappetent sheep although almost all sheep excreted salmonellas in faeces. Lesions of salmonellosis were found only in sheep that were seriously ill or had died, which suggested that, under the conditions of lot-feeding and sea transport, most sheep with enteric lesions are likely to die. Lesions were not found in feeding controls. Adrenal gland weights, an indicator of stress duration and severity, were used to examine the temporal sequence of events in the development of salmonellosis. Weights were greater in sheep that died of inanition than in controls, suggesting that inappetent sheep were already stressed for that reason. The presence of enteric lesions of salmonellosis was associated with further increases in adrenal gland weights.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240172 TI - Detection of bicarbonate administration (milkshake) in standardbred horses. AB - Total plasma carbon dioxide (TCO2) concentrations were measured in Standardbred horses to determine criteria to discriminate between normal horses and horses with excessive TCO2 concentrations on raceday. TCO2 concentrations from stabled horses were distributed normally with a mean of 30.2 mmol/L and a standard deviation of 1.2 (n = 192) while pre-race TCO2 concentrations were not normally distributed. The results indicate that about 50 horses per million are likely to have TCO2 concentrations greater than or equal to 35 mmol/L and that it is extremely unlikely that a normal horse would have a resting TCO2 concentration above 36 mmol/L. These values were associated with sensitivities of 67% and 59%, respectively, and with a specificity of 100%. TCO2 concentrations were relatively stable in blood samples stored at 4 degrees C for 4 days, whereas the TCO2 in specimens stored at room temperature (25 degrees C) and at ambient temperature (16-28 degrees C) declined progressively over 5 days. The accuracy and precision of the Beckman EL-ISE Auto Analyser were acceptable and within the manufacturers specified range. Paired specimens analysed using a Beckman EL-ISE Auto Analyser and a Kodak Dry Chemistry Analyser were not significantly different. However, the measurements made using the Kodak Dry Chemistry Analyser averaged 0.5 mmol/L higher than those analysed on the Beckman EL-ISE. The significance of these sources of variation in TCO2 concentration in relation to the testing of horses for 'milkshake' administration are discussed. PMID- 8240173 TI - Age-related changes in plasma biochemical values of farmed emus (Dromaius novaehollandiae). AB - Blood samples were collected from 40 emus (Dromaius novaehollandiae) of 4 different age groups ranging from 1 week to 14 months. Plasma values of glucose, cholesterol, uric acid, total protein, albumin, creatine kinase, aspartate amino transferase, alkaline phosphatase, calcium, phosphorus and magnesium were measured. Fourteen-month-old birds had lower plasma glucose values and enzyme activities and higher plasma protein values than younger birds. One-week-old birds had higher cholesterol and uric acid values than other age groups. Plasma calcium, phosphorus and magnesium values did not differ across the age profiles sampled. PMID- 8240174 TI - The medium frequency (7.5 MHz) ultrasound image characteristics of cattle skin. AB - The medium frequency (7.5 MHz) ultrasound images of cattle skin, as a model for other domestic animals, are described and compared with the published information for human skin. Although some difficulties are apparent in identifying the skin surface and therefore measuring tissue thicknesses, the papillary and reticular layers of cattle skin, about 2 and 4 mm thick, respectively, can be differentiated using a water standoff with a vegetable oil coupling medium. The difference in echo intensities and patterns between these dermal components supports the concept that ultrasound tissue characterisation using higher frequencies may prove to be a useful non-invasive procedure in livestock production and veterinary science. However, a detailed protocol is required. Care is needed in the choice of ultrasound coupling medium for good resolution of cattle skin. Higher frequency ultrasound equipment would be necessary for imaging the skin of small animals. Potential applications of this new technology are discussed. PMID- 8240175 TI - Field observations in Israel on hypersensitivity in cattle, sheep and donkeys caused by Culicoides. AB - Hypersensitive lesions in the skin of sheep, cattle and donkeys from various regions in Israel are described. No cause other than insect bites was found. The animals were free of ticks, lice, fleas and blackflies, and of all the prevailing insect groups; Culicoides spp were considered as the most probable cause of the hypersensitivity. Lesions on the legs and abdomen were attributed mainly to C obsoletus Meigen, C schultzel gp and C puncticollis Becker, and less to C imicola Kieffer. Lesions on the back, flanks, neck and head were attributed mainly to C imicola, less to C newsteadi Austen and rarely to C pulicaris Linnaeus. PMID- 8240176 TI - An evaluation of a live Toxoplasma gondii vaccine in Tammar wallabies (Macropus eugenii) PMID- 8240177 TI - Suppurative otitis in cattle associated with ear mites (Raillietia auris). PMID- 8240178 TI - Pilot survey for Lyme disease antibodies in Brisbane dogs. PMID- 8240179 TI - Ivermectin-resistant Haemonchus contortus in Australia. PMID- 8240180 TI - Ocular complications of PUVA therapy. AB - PUVA, the combination of psoralen and long wave ultraviolet radiation is widely used in the management of psoriasis, vitiligo and several other dermatological disorders. The potential for long term treatment to cause ocular damage remains to be determined and despite the large numbers of patients who have received PUVA treatment, development of cataract is exceedingly rare. This paper discusses cataract formation, reviews the literature concerning the ocular complications of PUVA therapy and proposes guidelines for ocular protection during photochemotherapy. PMID- 8240181 TI - Interferons for cutaneous T-cell lymphoma monotherapy or combination with PUVA. PMID- 8240182 TI - Paraneoplastic pyoderma gangrenosum. AB - Pyoderma Gangrenosum (PG) is often associated with an underlying disease. PG as a paraneoplastic disease is illustrated by the presentation of four patients with malignancy of myeloproliferative origin and PG. An associated malignancy is found in approximately 7% of patients with PG, most commonly haematologic in nature and in particular leukaemia. Clinically the PG is often of the superficial bullous variant and is associated with a poor prognosis. PMID- 8240183 TI - Two cases of onychomycosis due to Candida zeylanoides. AB - As the range of yeasts and filamentous fungi reported to cause onychomycosis has increased, uncommon genera or species have appeared in the literature. Candida zeylanoides is an example of a yeast rarely involved in human pathology. Between 1989 and 1991, two cases of onychomycosis due to this yeast were encountered in Wollongong, NSW. There were similarities in the cases, as both involved finger- or thumb-nails and both patients were female (one sixty years of age and the other, seventy-two). Evidence of pathogenicity is presented and subsequent topical treatment was successful in both cases. PMID- 8240184 TI - Treatment of eumycetoma with ketoconazole. AB - Ten patients with eumycetoma were treated with oral ketoconazole in the dosage of 400mg/day for 8 to 24 months. In eight cases the foot was affected: four were due to Madurella mycetomatis and one each due to M grisea, Pyrenochaeta romeroi, Acremonium kiliense and A falciorme. One mycetoma which affected the back and perineum was due to A kiliense, and one case presented with multiple sebaceous cysts and the scalp and M mycetomatis was isolated from the lesion. Clinical and laboratory tests showed excellent tolerance to the drug, with no adverse reactions. Complete cure was obtained in six patients and two showed good responses. The cured patients were followed up for a period ranging from three months to two years without any evidence of recurrence. PMID- 8240185 TI - Sweet's syndrome and chlamydial infection. PMID- 8240186 TI - Post-kala-azar dermal leishmaniasis. PMID- 8240187 TI - Treatment of basal cell carcinoma with intralesional interferon. PMID- 8240188 TI - Localised recurrent painful bruises. Gardner-Diamond syndrome of autoerythrocyte sensitisation, psychogenic purpura. PMID- 8240189 TI - Increasing importance of Trichophyton tonsurans in childhood tinea in New South Wales. The pattern of childhood tinea in New South Wales, Australia 1979-1988: the emergence of Trichophyton tonsurans as an important pathogen in tinea capitis in white children. AB - A retrospective study was undertaken of 192 cases of culture proven tinea capitis occurring between 1979 and 1988 in a predominantly white population of children in New South Wales (NSW). The aim of the study was to identify whether Trichophyton tonsurans was increasing in importance in the causation of this condition in Australia as it was in many other parts of the world. The results of our study were compared with those of earlier Australian studies. Our study demonstrated that Trichophyton tonsurans is now equal in importance to Microsporum canis which was previously the strongly predominant organism. PMID- 8240190 TI - Etretinate therapy of solar-related kerato acanthoma. AB - Nineteen patients with a total of 22 keratoacanthomas were treated in this trial with 1 mgm/kg/day of etretinate for six weeks. The patients had predominantly solitary keratoacanthoma, all associated with severe solar damage. There was a provision for reduction of dose and also for extension of length of treatment time for lesions which had responded but had not completely disappeared at the end of six weeks. Sixteen of the nineteen patients cleared without recurrence. Two patients had therapy stopped because of side effects and in these, further treatment was required. These side effects were severe paronychia and elevation of serum lipids. Other side effects were almost exclusively muco-cutaneous. Keratoacanthomas on the head, neck and upper limbs had cleared completely at the end of six weeks therapy, with a variable degree of ridged scarring. Lesions on the lower legs almost invariably required treatment times which were longer than six weeks and could be up to twelve weeks. Etretinate therapy of solar related keratoacanthoma should be considered for large lesions, multiple lesions, lesions in areas which are difficult to treat, and lesions in patients unsuited for destructive therapy or surgery. PMID- 8240191 TI - Situational awareness: the trainability of the near-threshold information acquisition dimension. AB - Ten subjects participated in training to identify abstract visual stimuli under three successively more difficult protocols where the stimuli (targets) were small, and target-on time was 67 ms (2,100 trials) or 33 ms (2,100 trials). Joysticks and push buttons were used for responses. The protocols were target detection, recognition, and identification. Backward masking was used to control visual access time. The experimental design was repeated measurements, each subject as his own control. Subjects exceeded 95% correct responses early under the 67 ms target-on time condition. For the 33 ms condition, most subjects achieved between 70 and 95% correct responses at the end of the training. Differences between protocol challenges at 33 ms were significant (at or beyond p < 0.01). The outcome of this study is consistent with our concept of situational awareness described in our overview paper. PMID- 8240192 TI - Body volume changes during simulated microgravity I: Technique and comparison of men and women during horizontal bed rest. AB - It has been hypothesized that many of the adaptive processes that occur in man during exposure to microgravity involve the rapid headward shift of fluids during the first few hours of spaceflight. Impedance plethysmography was used to measure lower body volume changes in men and women during horizontal bed rest simulated microgravity. Segmental volumes calculated from the leg impedance values are compared to similar volumes determined from detailed anthropometric measurements. The results of this experiment indicate that men and women respond differently to bed rest and that impedance plethysmography can be used to quantify the extent of fluid redistribution in the body during actual or simulated microgravity in a manner that corresponds well with anthropometric determinations. PMID- 8240193 TI - Body volume changes during simulated microgravity. II: Comparison of horizontal and head-down bed rest. AB - Changes in segmental fluid volume were measured in 10 male subjects during 7 d of either horizontal (0 degrees) or antiorthostatic (-6 degrees) head-down bed rest as part of the Joint U.S./U.S.S.R. Hypokinesia Program. Impedance plethysmography (IPG) was used to measure leg, pelvic, thoracic, and arm volume changes prior to, during, and following bed rest. Results of this study demonstrate that antiorthostatic bed rest produces a larger and more persistent thoracic fluid shift than horizontal bed rest. Segmental volume responses found during this investigation are compared to those obtained during other simulated or actual space-flight experiments. Antiorthostatic thoracic volume changes from this study were found to closely approximate those estimated to take place in space while leg volume changes produced by either 0 degrees or -6 degrees bed rest were much less than those observed by others in the microgravity environment. PMID- 8240194 TI - Seven-day pyridostigmine administration and thermoregulation during rest and exercise in dry heat. AB - Seven men participated in a double-blind study of effects of multiple-dose oral pyridostigmine bromide (PB) on physiological responses to 4-h heat stress tests (HST's) in a hot dry environment, 42 degrees C, 20% relative humidity. Subjects underwent 2 7-d series of tests, separated by 72 h, taking 30 mg PB every 8 h in one series, and placebo in the other. Each HST began right after the 0800 dose of PB or placebo. Subjects drank ad libitum during each HST, and performed two 55 min treadmill walks at about 40% VO2max during the last 2 h. Inhibition of red cell cholinesterase at the start of exercise averaged 30.0% in subjects taking PB, and did not differ significantly among HST's with PB. PB increased sweating and evaporative water loss by about 4%, and lowered chest skin temperature during exercise by 0.7 degrees C; but it had no significant effect on rectal temperature, other skin temperatures, O2 uptake, or fluid balance. PB alone had no significant effect on heart rate (HR), but had a significant interaction with day: although PB had essentially no effect on HR in the 1st HST, its effect increased progressively so that HR during exercise in the 4th HST was 8 beats.min 1 lower with PB. Multiple-dose PB had only slight effects on responses to moderate exercise-heat stress beyond those described after single-dose PB, and we found no adverse effects of multiple-dose PB administration. PMID- 8240195 TI - Postural and performance changes following exposures to flight simulators. AB - Flight simulators are cost-effective, safe, and flexible training tools for aviators. However, their advantages may be offset by the occurrence of motion sickness-like symptoms which have been reported during and following simulator training. Although symptoms have been well documented in simulators, their time course, causes, and implications for training are not yet fully understood. Tests of standing and walking steadiness were administered along with cognitive and motor performance tests to Navy and Marine Corps aviators before and after their regular simulator training, resulting in records of 726 pilot exposure observations. When exposed pilots were compared to a control group who did not fly in a simulator, statistically significant decrements in postural equilibrium test scores were found for all of the moving base simulators, but for only one of the three fixed-base simulators. The size of these losses was approximately 15% of baseline. Cognitive and motor changes, while statistically significant, were complicated by learning effects in all groups for all tests. When compared to the control group, improvement in cognitive scores was always less in the simulator groups, but greater improvement occurred in the simulator groups for motor speed scores, although this latter difference was small (< 1% of baseline). Further study of performance changes is recommended. In those simulators where significant effects occurred, the ataxia and the time-course of the disequilibrium should be followed. PMID- 8240196 TI - Autonomic function and plasma catecholamines following stressful sensory stimuli. AB - This experimentation defined a limited role for epinephrine in the autonomic nervous system function and the nausea that occurred following motion sickness testing. Individual responses to stressful sensory stimuli and nausea, as reflected by rising peripheral levels of epinephrine, were not significantly diminished upon repeated exposure and adaptation to the stressor. However, subjects who demonstrated more robust elevations of epinephrine in response to nausea displayed higher resistances to stressful motion stimuli. Modulation of peripheral catecholaminergic function with dexamethasone, or scopolamine plus amphetamine, suggested that altered autonomic nervous system function and nausea following motion sickness testing were not mediated by peripheral catecholamine receptor stimulation. Marked differences were noted in individual responses to drug and systemic responses of epinephrine and norepinephrine. It is possible that responses in epinephrine to motion sickness testing may predict resistance to stressful motion, and represent a peripheral manifestation of some as yet unknown central event of etiologic relevance. PMID- 8240197 TI - Pressurization and acute mountain sickness. AB - Numerous cases of acute mountain sickness (AMS) during trekking were reported to have been successfully treated with portable pressure chambers. The effect of early pressurization during acute altitude exposure in the Alps had not been previously studied. In order to test the hypothesis that an early pressurization of unacclimatized subjects for 3 h could prevent or delay the appearance of symptoms of AMS, 51 previously healthy subjects climbed from 1,030 to 4,360 m within 12 h. Upon arrival at 4,360 m, AMS scores (Lake Louise Consensus Questionnaire '91), oxygen saturation (SaO2), and heart rate (HR) were determined at rest. The subjects were then randomly divided in two groups; one group was pressurized to 200 mBar for 3 h while the other rested. AMS score, HR, and SaO2 were similar in both groups before treatment. AMS score had decreased (from 2.44 +/- 0.41 (S.E.) to 0.89 +/- 0.26, p < 0.05) and SaO2 had increased (from 75.22 +/ 1.32% to 79.07 +/- 1.27%, p < 0.05) in the treatment group 15 min after leaving the pressure chamber whereas the control group had unchanged AMS score (2.50 +/- 0.40 vs. 2.40 +/- 0.40, N.S.) and SaO2 (77.83 +/- 1.41 vs. 76.67 +/- 1.24, N.S.). The next morning, however, AMS score, HR, and SaO2 were similar for both groups. It is concluded that during acute ascent in the Alps, an early 3-h pressurization of unacclimatized subjects does slightly delay the onset of AMS but does not prevent the illness nor does it attenuate its severity upon appearance. PMID- 8240198 TI - Aphakia and artificial lens implants in the civil airman population. AB - The Federal Aviation Administration allows civilian airmen with aphakia to fly with waivered certificates. This study analyzes, in the civil airman population, the distribution of aphakia and intraocular lens (I.O.L.) implant by type (unilateral, bilateral), class of airman medical certificate, and gender, for a 4 year period (1982-85). Medical records were individually evaluated for all certified airmen who were carrying FAA-specific codes for the pathology categories during the study period. The prevalence of both aphakia and I.O.L. increased most for bilateral type and second-class certificate holders. During the study period, the prevalence of aphakia increased most for males and I.O.L. increased most for females. The incidence declined in both pathology categories during the later years of the study period. Implications for aeromedical certification of the increasing prevalence of aphakia and I.O.L. in the civil airman population are discussed. The modification of surgical procedures for cataract extraction and evolution of I.O.L. devices strongly suggests continued specialized aeromedical certification and clinical research review. PMID- 8240199 TI - Catapult launch-associated cardioversion of atrial fibrillation. AB - A patient is presented with new-onset atrial fibrillation. While being air evacuated, he cardioverted during the acceleration associated with aircraft carrier catapult launch. This case suggests a possible myocardial response to the kinetic energy produced by the acceleration force, similar to the electrical energy commonly used for cardioversion. Cardioversion using acceleration forces is probably not a clinically useful modality; however, this case demonstrates the importance of acceleration forces on patients during medical evacuation from aircraft carriers. PMID- 8240200 TI - The wartime need for aeromedical evacuation physicians: the U.S. Air Force experience during Operation Desert Storm. AB - Air transportation has been the primary method of moving patients by the armed services of the United States since 1949. It is fast, reliable, and allows for centralized medical care. Aeromedical Evacuation (AE), performed by the U.S. Air Force under Department of Defense directive, was intended as a method to transport medically stable patients. Modern warfare has evolved into a process capable of generating large numbers of casualties in a short period of time that can overwhelm local medical facilities. Such casualties would then require immediate transportation in order to obtain appropriate treatment. The terrorist bombing of the U.S. Marine barracks in Beirut and the 1989 military action in Panama (Operation Just Cause) are recent experiences where unstable casualties were transported by an AE system not designed to care for acute injuries while en route to definitive care. During Operation Desert Storm, Aeromedical Evacuation Flight Surgeons (AE/FS's) augmented AE crews and provided flexibility to transport critically ill patients. Future planning should augment designated AE crews with appropriately trained physicians and include equipment on aircraft to resuscitate patients that decompensate inflight. PMID- 8240201 TI - A mathematical model of G time-tolerance. AB - We propose a model to explain experimentally observed effects of Gz onset rates and levels on the time of occurrence of loss of vision and/or consciousness. The model is based on the existence of two generally accepted parameters: a G limit beyond which cerebral perfusion ceases, and a buffer time between loss of perfusion and loss of function. When applied to ramp onset G profiles, the model predicts a generally hyperbolic locus of endpoints, similar to the well-known Stoll curve, except for the dip. The advantage of the model is its applicability to any G onset profile. Data from the literature support the assumptions of the model and its results, including the absence of the dip in the locus for a ramp onset. The results call into question some concepts used to design G avoidance inflight strategies and the usefulness of some experimental centrifuge methods. The model may enable an increase in the accuracy of predictions of the time of visual or cerebral loss of function under various G profiles. PMID- 8240202 TI - The opticogravic nerve: eye-level anatomic relationships within the central nervous system. AB - The anatomic relationships between arterial blood supply and key structures within the central nervous system (CNS) are important in comprehending the neurophysiological effects of acceleration (+Gz) stress, including +Gz-induced loss of consciousness (G-LOC). An accurate understanding of the location of the vascular and neurologic structures at eye-level is vital, since it is possible to determine precisely when visual symptoms occur. Cerebral perfusion is supported by the pressure-equalizing effects provided by the cerebrospinal fluid. While brain and brain stem are protected by this pressure compensation, the eye is not. Decreased visual function results when retinal perfusion is compromised. G-LOC generally occurs following loss of visual function. The exact location(s) of altered perfusion within the CNS that results in G-LOC is currently unknown. To thoroughly understand G-LOC, it will be necessary to understand which CNS structures must be affected by +Gz-induced ischemia to cause G-LOC. Based on previous theoretical considerations of loss of consciousness as a protective mechanism, it is possible to consider the eye (visual system) as a dual sensor for both vision and gravitational (acceleration) stress. Due to this expanded definition of the sensory functions of the second cranial nerve, it would therefore be more appropriate to describe it as the "opticogravic" nerve. This manuscript discusses some of these considerations and the eye-level neuroanatomic relationships of vital importance for the acceleration medical subspecialist. PMID- 8240203 TI - Cases from the aerospace medicine residents' teaching file. Case #54. Fibrous dysplasia. PMID- 8240204 TI - Aircraft carrier psychiatric diagnoses. PMID- 8240205 TI - Standardized aircraft accident reporting. PMID- 8240206 TI - Metacarpal fracture and the failing aviator. PMID- 8240207 TI - Hyperbaric treatment on a lunar station. PMID- 8240208 TI - Women's health issues and space-based research. PMID- 8240209 TI - Invited commentary: cryopreservation as a tool for the study of selectively bred lines in rodent behavioral genetics. AB - Cryopreserved embryos should survive centuries. Upon rederivation, in vitro survival is high. Upon reimplantation, in vivo survival is substantial. Genetic stability of the embryos at -196 degrees C is theoretically absolute for several decades and likely high for centuries. Cryopreservation and rederivation services are available at a reasonable cost from commercial vendors. Preservation of a sufficient number of embryos can serve as an insurance policy against loss of the lines from several causes. In addition to the protective utility of a single banked sample, repeated preservation during the course of selection offers several advantages. Correlated responses to selection which appear late in selection may be verified by use of a cross-sectional experimental design to take a "snapshot" of the gradually developing response. After attainment of the selection limit, a generation may be chosen retrospectively which represents the best compromise between maximizing response and minimizing inbreeding irrelevant to the selected trait. Estimates of changes in gene frequencies could be derived from molecular biological analyses. An example of such an experiment in progress employing embryos from the COLD and HOT selected mouse lines is discussed. PMID- 8240210 TI - Familiality of female and male homosexuality. AB - We examined data from a large cohort of homosexual and heterosexual females and males concerning their siblings' sexual orientations. As in previous studies, both male and female homosexuality were familial. Homosexual females had an excess of homosexual brothers compared to heterosexual subjects, thus providing evidence that similar familial factors influence both male and female homosexuality. Furthermore, despite the large sample size, homosexual females and males did not differ significantly from each other in their proportions of either homosexual sisters or homosexual brothers. Thus, results were most consistent with the possibility that similar familial factors influence male and female sexual orientation. However, because results conflicted with those of some other studies, and because siblings' sexual orientations were obtained in a manner likely to yield more errors than in these other, smaller studies, further work is needed using large samples and more careful methods before the degree of cofamiliality of male and female homosexuality can be resolved definitively. We also examined whether some parental influences comprised shared environmental effects on sexual orientation. Scales attempting to measure such influences failed to distinguish subjects with homosexual siblings from subjects with only heterosexual siblings and, thus, did not appear to measure shared environmental determinants of sexual orientation. PMID- 8240211 TI - Sensation seeking in opposite-sex twins: an effect of prenatal hormones? AB - Intrauterine hormones and position with respect to male and female littermates influence sexually dimorphic adult behavior in litter-bearing animals. Opposite sex dizygotic twins offer the opportunity to examine analogous effects on sex related human behaviors. To illustrate this approach, Sensation Seeking Scale (SSS) scores from 422 British twin pairs, including 51 opposite-sex pairs (Zuckerman, M., et al., J. Consult. Clin. Psychol. 46:139-149, 1978), were reanalyzed. Zuckerman et al. (1978) have shown that some aspects of sensation seeking are consistently increased in males relative to females. In comparing age adjusted data for opposite and same-sex twins, our reanalysis demonstrated the predicted increase in sensation seeking in female members of opposite-sex pairs. Results were significant for measures of disinhibition, experience seeking, and overall sensation seeking. In contrast, male opposite-sex twins were not significantly different from male same-sex twins. Although psychosocial explanations of the increased sensation seeking in opposite-sex female twins cannot be excluded, these finding are consistent with hypothesized in utero hormonal influences on later behavioral development. PMID- 8240212 TI - Differences in heritability across groups differing in ability, revisited. AB - Three recent studies have used twin data to explore the possibility of differential contributions of heritability and environmentality to individual differences in cognitive ability as a function of ability level (Detterman, D. K., et al., Behav. Genet. 20:369-384; 1990; Bailey, M. J. and Revelle, W., Behav. Genet. 21:397-404, 1991; Cherny, S. S., et al., Behav. Genet. 22:153-162, 1992). All arrived at different conclusions: higher heritability at the low end, higher heritability at the high end, and no differential influence, respectively. The current report involves a sample of 148 identical and 135 fraternal twin pairs from the Western Twin Project who were tested on a battery of intelligence and achievement tests to further explore the issue. The results suggest no significant differences in heritability at either the high or the low end, although a trend toward higher heritability for children of higher ability is evident. Individual differences for a composite ability/achievement score showed significantly greater influence of shared family environment at the low end than the rest of the distribution. In general, results for cognitive ability and academic achievement were highly similar. PMID- 8240213 TI - Is the gender difference in mental rotation disappearing? AB - Several investigators have used meta-analysis to compare the results of studies of gender differences on various spatial tests and have concluded that the magnitude of the gender difference in spatial ability is decreasing over time. The present study used meta-analytic techniques to compare the effect size (d) of the gender difference in 14 studies published from 1975 to 1992 which administered the Mental Rotations test to adolescents and young adults. Males scored significantly higher than females in all the studies. Analyses of the d's computed for the studies revealed that the magnitude of the gender difference on the Mental Rotations test has remained stable over time. Neither the Pearson correlation relating the d's to the publication dates of the studies nor the Z test of the linear contrast relating the publication dates of the studies to the effect sizes showed a linear change in the size of the gender difference over time. The finding of a stable gender difference on the Mental Rotations test argues against the general conclusion that the gender difference in spatial ability is decreasing. PMID- 8240214 TI - Inbreeding depression and intelligence quotient among north Indian children. AB - This study presents the assessment of inbreeding depression on the intelligence quotient among north Indian Muslim Children of school age. The Wechsler Intelligence Scale for Children (WISC-R)-74 was given to the children in both groups (50 each non-inbred and inbred of the first-cousin status), aged 6 to 11 years and from the same socio-economic status. The change of the mean follows genetic theory; however, the nature of the change in variance seems to be somewhat different. PMID- 8240215 TI - Male mating speed in Drosophila melanogaster: differences in genetic architecture and in relative performance according to female genotype. AB - From a critical review of the literature on mating speed in Drosophila, the importance of fast mating in male fitness is questioned. The genetic architecture of male mating speed (MMS) has been evaluated in D. melanogaster through a populational analysis and a full 5 x 5 diallel cross between inbred lines. The results emphasize the fundamental role of the female genotype in both the absolute and the relative MMS performances. Somewhat different genetic architectures for MMS are revealed according to the female used in the tests. It is suggested that different parts of the complex genetic system involved in the male's "behavioral sexual phenotype" are relevant depending on the female's characteristics, thus causing the heterogeneity in the MMS genetic architecture. An overall picture reveals a genetic system characterized by additivity, dominance for fast mating, and no influence of the X chromosome. There results do not support strong natural selection favoring fast mating in Drosophila males. PMID- 8240216 TI - Sources of behavioral deviation modeled by early color preferences in quail. III. Developmental stability and normative canalization. AB - Successful artificial selection of quail (C. coturnix japonica) for divergent early approach preferences for, and imprintabilities to, the wavelength characteristics of otherwise identical visual stimuli decreased the trait's developmental stability. The developmental error that resulted from selecting for extreme initial preferences was associated with enhanced imprintabilities. Conversely, the developmental error that resulted from selecting for extreme imprintabilities was associated with enhanced expression of directional gene effects in extreme initial preferences. The data are attributed to selectively relaxed normative canalization of trait development, and to related enhanced expression of genetic, environmental, and stochastic influences in trait deviations. PMID- 8240217 TI - Inheritance of predisposition to catalepsy in mice. AB - "Pinch-induced" catalepsy (excessive freezing) was studied in mice of nine inbred strains. The CBA/Icg strain had the highest predisposition to catalepsy, with 56% males and 73% females exhibiting 120-s immobility after four tests. A Mendelian analysis of predisposition to catalepsy was performed on CBA/Icg and AKR/J mice contrasting in this trait. Reciprocal F1 hybrids did not display freezing after 10 tests. The number of cataleptic and noncataleptic animals among the F2's and in the families of the CBA x F2 backcrosses agreed with the hypothesis that the pronounced predisposition to catalepsy in the CBA/Icg mice was determined by a single autosomal locus in homozygous recessive condition. PMID- 8240218 TI - Lack of strain differences in spatial learning among seizure-prone and seizure resistant mice. AB - Learning rates were examined in the following inbred mice strains: DBA/2, C3H/He, C57B1/6J, E1, and ddY. DBA/2 mice become susceptible to audiogenic seizures after 2-3 weeks of age and E1 mice have generalized seizures in response to handling after 3 months of age, but the remaining three strains do not develop seizures. In this study, mice from all five strains underwent 32 training trials in a Morris water maze at 7-9 weeks of age. The seizure-prone DBA/2 and E1 mice, along with the nonepileptic ddY and C57B1/6J mice, exhibited learning at similar rates, but the nonepileptic C3H/He mice were unable to learn the water maze task, probably due to visual difficulties. In the C57B1/6J strain only, female mice learned the task significantly faster than males. There was no difference in the learning rate between the E1 strain and its parent ddY strain, or any correlation between spatial learning ability and kindling rates in these strains. PMID- 8240219 TI - Genetic analysis of isolation-induced aggression. II. Postnatal environmental influences in AB mice. AB - Recently, we reported on two closely related inbred mouse strains, ABG and AB//Halle, that display extreme differences in isolation-induced intermale aggression. In the present article we investigated the influence of both maternal and social postnatal environmental influences. No effects were found of the postnatal maternal environment. Likewise, whether animals after weaning were housed together in same-strain or mixed-strain groups did not influence their subsequent aggressive behavior. We conclude that the aggressive behavior of ABG and AB//Halle is rather robust with regard to postnatal environmental modification and that the difference between the two strains is most likely due to only few genetic factors. PMID- 8240220 TI - Elevated dark-adapted thresholds in hypopigmented mice measured with a water maze screening apparatus. AB - In previous electrophysiological experiments from hypopigmented animals (mice, rats, rabbits), single-unit recordings from both retinal ganglion axons and cells in the superior colliculus have demonstrated an increase in threshold in the dark adapted state which is roughly proportional to the ocular melanin concentration. In the present study we compared an albino mouse strain which is relatively resistant to light damage and the beige mouse mutant to their wild-type controls in a situation that involved unanesthetized, unrestrained mice as a control to the electrophysiological single unit experiments. We used a six-chambered water maze. Animals were trained to swim to an illuminated ramp until their performances leveled off (about 10 days). The animals were then dark-adapted for 24 h and tested after reducing the luminance level of the water maze. We found that the albino mice failed to find the ramp when the luminance fell to 1.58 x 10(-3) cd/m2 (p < or = .0001), the beige mice failed at 2.00 x 10(-4) cd/m2 (p < or = .0001), and the normally pigmented controls performed to 5.00 x 10(-5) cd/m2 (p < or = .0001). These results support our previous findings that the sensitivity defect in hypopigmented animals is proportional to the degree of ocular hypopigmentation. PMID- 8240221 TI - Learning in inbred mice: strain-specific abilities across three radial maze problems. AB - Mice belonging to the C57BL/6, DBA/2 (DBA), and C3H/He (C3H) strains were compared in three different eight-arm radial maze tasks requiring various degrees of spatial and nonspatial information processing. The results show that, on the standard radial maze task, C57 performed better than DBA, which, in turn, performed better than C3H. Fewer differences in the four-baited arm task and no difference in the cued version task were found between C57 and DBA, while C3H still performed more poorly. The high performance shown by C57 mice in all problems seems to be related to their ability to build up maze-running patterns based upon an optimal proportion of 45 degrees angle turns, according to the demand of the situation. The cognitive and discriminative mechanisms involved in the solving of each task, the sensorial characteristics of the three strains, and the limits of an approach based upon neuroanatomical-behavioral correlations are discussed. PMID- 8240222 TI - Genetic and behavioral tests of the McManus hypothesis relating response to selection for lateralization of handedness in mice to degree of heterozygosity. AB - McManus advanced a genetic hypothesis to explain differences of lateralization between HI and LO lines of mice selectively bred for degree of handedness. It states that lateralization is a function of heterozygosity. Specifically it predicts that (a) the HI line will be more heterozygous than the LO line and (b) populations with a greater average heterozygosity (AH) will be more strongly lateralized. Both genetic and behavioral predictions were tested here. Results using coat color and biochemical variants show that AH in the HI line is somewhat less (not greater) than that in the LO line. The handedness of HET control mice and HI by LO reciprocal hybrids, where AH is greater than that of the HI line, exhibits lessened (not greater) lateralization. Results reject the heterozygosity hypothesis. A model for the inheritance of human handedness that accounts for difficulty in detecting heritable differences in degree of asymmetry is presented. PMID- 8240223 TI - Release kinetics and correlation with hemodynamic dysfunction of cardiac troponin T in coronary effluent from isolated rat hearts during reperfusion. AB - Previously, we reported that cardiac troponin T (TnT) can be detected and measured in coronary effluent from isolated rat hearts during hypoxia. The present study was designed to evaluate the release kinetics of TnT from post ischemic rat hearts. Using the Langendorff technique, the hearts were reperfused for 4 h after 20 min or 60 min of global ischemia. Coronary flow was measured by timing the collection of the coronary perfusate that dripped from the hearts, and left ventricular pressure (LVP) was monitored continuously during the experiments. The amount of TnT released in 1 min was compared with the release of creatine kinase (CK) and lactate dehydrogenase (LD). The release kinetics of CK and LD showed a monophasic pattern and the levels at 4 h after reperfusion returned to baseline levels. By contrast, the release kinetics of TnT showed a small peak followed by a larger and more sustained peak. There were good negative correlations between developed pressure of LVP and both sigma TnT and the amount of TnT released within 1 min at 4 h after reperfusion. These results indicate that the release kinetics of TnT is different from that of CK and LD during reperfusion, and further that cardiac TnT is a useful indicator of myocardial cell damage and can be used to evaluate the degree of myocardial cell damage in both the early and late phase of acute myocardial infarction. PMID- 8240224 TI - No evidence of malonyldialdehyde formation during reoxygenation injury in vitamin E-deficient rat heart. AB - Vitamin E is an endogenous antioxidant and is known to afford protection against lipid peroxidation. If lipid peroxidation was an important factor in the pathogenesis of reoxygenation injury in heart, then both the extent of lipid peroxidation and cell injury would be expected to be exacerbated in vitamin E deficient hearts. To study reoxygenation injury in the present experiments, rat hearts were perfused in the Langendorff mode with a modified Krebs-Henseleit buffer under anoxic conditions for 60 min before resuming normoxic perfusion for 20 min. Creatine phosphokinase (CPK) activity and malonyldialdehyde (MDA), a product of lipid peroxidation, were assayed in the perfusate effluent from hearts during reoxygenation injury. Also, myocardial MDA and vitamin E contents were measured in extracts of freeze-clamped heart tissue obtained immediately before and 2 min after reoxygenation. Experiments were performed on hearts from groups of weanling rats fed either a vitamin E-deficient or vitamin E-supplemented diet (50 I.U. vitamin E/kg) for 5 to 6 weeks. After 5 weeks, the myocardial vitamin-E content was 103.8 +/- 5.3 (n = 5) and 11.5 +/- 1.6 (n = 4) ng/mg protein (mean +/ SEM) in the vitamin E-supplemented and vitamin E-deficient groups respectively. Perfused hearts from both dietary groups showed a peak of enzyme release 2 to 3 min after the reintroduction of oxygen, and enzyme release from vitamin E deficient hearts was two-fold greater than enzyme release from vitamin E supplemented hearts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240225 TI - Postischemic dysfunction of the heart induced by small numbers of neutrophils via formation of hypochlorous acid. AB - The role of polymorphonuclear neutrophils (PMN) in the injury of the heart following ischemia and reperfusion is still controversial. The aim of this study was to investigate whether small numbers of PMN may cause myocardial dysfunction in an isolated system, how the resulting loss of function can be characterized and whether the formation of hypochlorous acid (HOCl) can be responsible for the PMN-mediated effect. Isolated working guinea pig hearts were subjected to a 90% reduction of coronary flow for 30 min, with or without intracoronary infusion of homologous PMN (approximately 1-2 x 10(5) cells/min, i.e. about 5-10% of normal blood count). This ischemia was followed by a 15 min reflow period in a non working ("Langendorff") mode before work was resumed. In hearts perfused only with buffer, post-hypoxic heart function recovered to 75-80% of the initial value. Inclusion of unstimulated PMN did not further attenuate cardiac function. However, cardiac output was decreased to 42% of the initial value, provided thrombin (0.3 U/ml) and H2O2 (10(-5) M) were also present, and the retained PMN (about 10% of those infused) were additionally stimulated during reflow by application of FMLP (10(-6) M for 1 min). In these instances, coronary flow at any time of the experiment and release of lactate or purines during ischemia and reflow did not differ significantly between hearts perfused with or without PMN. There was no substantial release of myoglobin in controls and in PMN-treated hearts. Inotropic stimulation of the hearts with noradrenaline or exogenous Ca2+ caused a sustained increase in contractile force. However, the response was significantly reduced in PMN-perfused hearts in comparison to control hearts. The myocardial contents of high-energy phosphates with and without inotropic stimulation proved to be identical irrespective of whether experiments had been performed in the absence or presence of PMN. A similar loss of myocardial function as mediated by PMN could be produced by infusing chemically generated hypochlorous acid (HOCl, 5 x 10(-7) M for 10 min). Strikingly, that portion of the infused HOCl which actually reacted with cardiac tissue was comparable to the amount shown to be generated by stimulating 10(6) PMN retained in the coronary system (about 7 nmoles). Supplementing the perfusate with the scavengers L methionine (10(-4) M) or uric acid (5 x 10(-4) M) prevented the attenuation of heart function provoked by PMN. The results indicate that small numbers of PMN, sufficiently activated, can depress cardiac function after 30 min of ischemia.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8240226 TI - Ventricular distension and diastolic coronary blood flow in the anaesthetized dog. AB - There appears to be no agreement as to whether or not an increase in diastolic left ventricular pressure and/or volume can cause a decrease in diastolic coronary blood flow. We investigated the problem in the anaesthetized dog using a flaccid freely distensible latex balloon inserted into the left ventricle with the animal on extracorporeal circulation and the coronary perfusion pressure constant at about 45 mm Hg. Maximal vasodilatation and suppression of autoregulation in coronary vasculature was obtained by the intracoronary infusion of dipyridamole (10-40 mg/h). Ventricular volume was changed in steps of 10 ml from 10 to 70 ml and back to 10 ml, whilst recording coronary blood flow and left ventricular pressure in the left circumflex coronary artery. Over a range of ventricular volumes from 20 to 50 ml and a concomitant rise in diastolic ventricular pressure to about 20 mm Hg there was no change in the diastolic coronary flow. Only when the ventricular volume was more than two times the control value (i.e. exceeded 50 ml) and left ventricular pressure was more than 20 mm Hg, was there a decrease in coronary flow. During the return of the volume to the control level there was a fall in diastolic flow and ventricular contractility with respect to the values obtained when the volume was increased; these two effects were transient lasting less than 10 min. It was not considered that any of the three models of the coronary circulation, waterfall, intramyocardial pump or varying elastance model could explain our results.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240227 TI - Acute alterations in systolic ventricular interdependence-mechanical dependence of right ventricle on left ventricle following acute alteration of right ventricular free wall. AB - The purpose of the study was to examine whether systolic ventricular interdependence can be acutely altered by changes in the mechanical properties of the ventricular wall. In eight acute canine studies, we released an aortic constriction during diastole. We measured right ventricular (RV) pressure changes (dPr) caused by sudden changes in left ventricular (LV) pressure (dPl). Measurements were obtained during control, 10 min after right coronary artery occlusion, and then 15 min after injecting glutaraldehyde into the RV free wall. By superimposing the pressure tracings of the beats immediately before and after the aortic release, the instantaneous pressure difference ratio (dPr/dPl) was calculated during systole. Maximal value of the pressure difference ratio decreased from control 0.11 +/- 0.04 to ischemia 0.08 +/- 0.03; (p < 0.05) and increased with glutaraldehyde 0.15 +/- 0.06; (p < 0.05). Thus, acute ischemia in RV free wall decreased the magnitude of systolic ventricular interdependence from LV to RV, while glutaraldehyde, which stiffens the RV free wall, increased the magnitude. PMID- 8240228 TI - Myocardial carnitine metabolism in congestive heart failure induced by incessant tachycardia. AB - Persistent tachycardia induces congestive heart failure (CHF), but the mechanism(s) of progressive ventricular dysfunction is (are) unclear. This study was designed to define possible metabolic causes of myocardial dysfunction in rapid ventricular pacing induced CHF. Twelve adult mongrel dogs were paced to 250 beats/min for 19 days. Plasma carnitine, norepinephrine and renin were measured at 0, 1, 2, and 3 weeks. Myocardial high energy phosphates, carnitine, glycogen, glucose, non-collagenous protein and collagen were measured at 19 days. Cardiac output, arterial pressure and pulmonary wedge pressure, measured at baseline and with CHF, showed a decrease in cardiac output and increase in pulmonary wedge pressure. Neurohumoral activation was evident by progressively increasing plasma norepinephrine and renin activity and depletion of myocardial norepinephrine. Plasma free carnitine rose significantly from 12.6 +/- 2.0 control to 28.3 +/- 3.8 nmol/ml at 19 days (p < 0.001), whereas myocardial total carnitine was lower in paced than in control dogs (6.0 +/- 1.9 vs. 14.1 +/- 3.5 nmol/mg non collagenous protein, p < 0.001). Myocardial ATP ATP and ADP were unchanged, while AMP decreased 22%, and creatine phosphate decreased 30% compared to control animals. Myocardial glucose was normal but glycogen was decreased 54% (p < 0.005). The low myocardial carnitine and elevated plasma carnitine in pacing induced CHF suggests altered carnitine transport or membrane integrity. PMID- 8240229 TI - Coexpression of alpha and beta myosin heavy-chain isoforms in atria of neonates and infants with congenital heart disease. AB - The relative amounts of cardiac myosin heavy-chain isoforms (MyHC) in right atrial tissue (RA) of 16 neonates and children suffering from congenital heart disease have been investigated. Quantification of MyHC was based on one dimensional gel electrophoresis and on histometrical evaluation of cyro-sections stained with monoclonal antibodies against alpha- and beta-MyHC. The mean right atrial pressures ranged from 2 to 14 Hg. The RA load was normal in eight patients (5.1 +/- 1.3 mm Hg) and overloaded in eight cases (10 +/- 2.5 mm Hg). The arterial oxygen saturation was normal in 12 and ranged between 85% and 89% in four cases. In all patients a large proportion of atrial myocytes coexpressed alpha- and beta-MyHC. However, in the cases with pressure overloaded RA the amount of beta-MyHC was found to be 1.6 times higher than in the cases with normal pressure. This indicates an adaptational response to overload, as was previously described for the adult human heart. In light of this finding, it seems important to conserve as much as possible of the trained right atrial wall during a Fontan type of operation. PMID- 8240230 TI - The glucose transporter family: structure, function and tissue-specific expression. PMID- 8240231 TI - 9-cis-retinoic acid is a natural antagonist for the retinoic acid receptor response pathway. AB - The pleiotropic activities of retinoids are mediated by two types of nuclear receptors, the retinoic acid receptors (RARs) and the retinoid X receptors (RXRs). All-trans-retinoic acid (RA) transcriptionally activates RARs, but not RXRs, whereas its natural stereoisomer, 9-cis-RA, is the ligand for RXRs. Here, we demonstrate that 9-cis-RA did not transcriptionally activate RARs, whereas in the presence of all-trans-RA the transactivation of RARs was inhibited in a dose dependent manner by 9-cis-RA. RAR homodimer complexes were destabilized in vitro in the presence of 9-cis-RA. This suggests that 9-cis-RA may be a natural antagonist of all-trans-RA for binding to RAR complexes. The levels of 9-cis-RA may determine by which pathway the transcription of retinoid-responsive genes is modulated. PMID- 8240232 TI - Molecular cloning and deduced amino acid sequences of the gamma-subunits of rat and monkey NAD(+)-isocitrate dehydrogenases. AB - A 600 bp cDNA fragment encoding part of the gamma-subunit of pig heart NAD(+) isocitrate dehydrogenase (ICDH gamma) was amplified by PCR using redundant oligonucleotide primers based on partial peptide sequence data [Huang and Colman (1990) Biochemistry 29, 8266-8273]. This PCR fragment was then used as a probe to isolate clones encoding the complete mature forms of the gamma-subunit from rat epididymis and monkey testis cDNA libraries. Comparison of the deduced amino acid sequences of the rat and monkey subunits and the partial sequence of the pig heart enzyme revealed a remarkably high level of sequence identity. The relationship between the deduced amino acid sequences of the NAD(+)-ICDH gamma subunits and those of nonmammalian NAD(+)- and NADP(+)-ICDH subunits is discussed. PMID- 8240233 TI - Mitogen-activated protein kinase (MAP kinase), MAP kinase kinase and c-Mos stimulate glucose transport in Xenopus oocytes. AB - Mitogens and growth factors acutely stimulate glucose transport in all cells to supply energy for their growth and division, but little is known about the signalling mechanism by which these agonists promote sugar uptake. Here we show that the transport of deoxyglucose and 3-O-methylglucose into Xenopus laevis oocytes is stimulated about 2.5-fold when mitogen-activated protein kinase (MAP kinase) is microinjected into these oocytes. We also demonstrate that microinjection of the proto-oncogene product c-Mos (an activator of MAP kinase kinase, which activates MAP kinase in Xenopus oocytes), and purified MAP kinase kinase produce similar increases in deoxyglucose transport. Since the activation of MAP kinase is a general response to almost all mitogens and growth factors, we propose that one of its downstream effects is the stimulation of glucose transport activity. PMID- 8240234 TI - Vectorial Ca2+ flux from the extracellular space to the endoplasmic reticulum via a restricted cytoplasmic compartment regulates inositol 1,4,5-trisphosphate stimulated Ca2+ release from internal stores in vascular endothelial cells. AB - Depletion of the Ins(1,4,5)P3-sensitive intracellular Ca2+ store of vascular endothelial cells after selective inhibition of the endoplasmic-reticulum (ER) Ca2+ pump by thapsigargin or 2,5-di-t-butylhydroquinone (BHQ) increases Ca2+ influx from the extracellular space in the absence of phosphoinositide hydrolysis. One model to account for these results suggests a close association between the internal store and the plasmalemma, allowing for the vectorial movement of Ca2+ from the extracellular space to the ER. Furthermore, recent evidence suggests that Ins(1,4,5)P3-induced Ca2+ release from intracellular stores is regulated by the free cytosolic Ca2+ concentration ([Ca2+]i). Thus agonist-induced Ca2+ entry may directly regulate Ca2+ release from internal stores. To test these hypotheses, we examined the effect of 1-(beta-[3-(4 methoxyphenyl)propoxy]-4-methoxyphenethyl)-1H-imidazole (SKF 96365), an inhibitor of Ca2+ influx, on unidirectional 45Ca2+ efflux (i.e. retrograde radioisotope flux via the influx pathway) and on [Ca2+]i as measured by fura-2. Bradykinin produced a transient increase in [Ca2+]i, reflecting release of Ca2+ from internal stores, and a sustained increase indicative of Ca2+ influx. In the absence of agonist, 45Ca2+ efflux was slow and monoexponential with time. Addition of BK dramatically increased 45Ca2+ efflux; 50-60% of the 45Ca2+ associated with the cell monolayer was released within 2 min after addition of bradykinin. Both the bradykinin-induced change in [Ca2+]i and the stimulation of 45Ca2+ efflux was completely blocked by loading the cells with the Ca2+ chelator BAPTA. At a supermaximal concentration of bradykinin (50 nM), SKF 96365 (50 microM) inhibited the rise in [Ca2+]i attributed to influx without affecting release from internal stores. At a threshold concentration of bradykinin (2 nM), SKF 96365 blocked influx, but stimulated Ca2+ release from internal stores, as indicated by increases in both the transient component of the fura-2 response and 45Ca2+ efflux. Thapsigargin (200 nM) and BHQ (10 microM) produced an increase in 45Ca2+ efflux that was completely blocked by SKF 96365 or by cytosolic loading with BAPTA. These results suggest the existence of a restricted sub-plasmalemmal space that is defined by an area of surface membrane which contains the Ca(2+) influx pathway but is devoid of Ca2+ pumps, and by a section of ER that is rich in thapsigargin-sensitive Ca(2+)-pump units.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8240235 TI - The Alzheimer beta-amyloid protein precursor/protease nexin-II is cleaved by secretase in a trans-Golgi secretory compartment in human neuroglioma cells. AB - Alzheimer beta-amyloid protein precursor (beta APP) is expressed endogenously and abundantly by human neuroglioma (H4) cells. Its secretory processing has been shown to involve discrete proteolysis within the beta A4 region, thus preventing beta-amyloid formation, by an enzyme which has been referred to as 'beta APP secretase'. This cleavage results in secretion of a soluble N-terminal 135 kDa protein and retention of an integral membrane C-terminal fragment within the cell. The membrane-associated C-terminal fragment is sorted to lysosomes where it undergoes limited degradation. We show here that most newly synthesized beta APP is degraded via a non-lysosomal pathway before maturation in H4 cells, and most mature beta APP is processed predominantly by the so-called secretase. The rapid kinetics of appearance/disappearance of a cleaved 135 kDa protein within a microsomal fraction and the slow accumulation of this form in the extracellular medium indicated that secretase cleaves beta APP in an intracellular compartment. Low-temperature block (20 degrees C) was used to demonstrate that beta APP is cleaved within a late Golgi compartment after sulphation which occurs in the trans-Golgi network (TGN). This is consistent with (1) the immunolocalization of most of the beta APP within a Golgi compartment that reacts with wheat germ agglutinin, (2) the fact that less than 1.5% of the total mature full-length beta APP is present at the plasma membrane and (3) subcellular fractionation studies which showed that the mature full-length and intracellular cleaved beta APPs co sediment with a membrane fraction that is slightly more dense than the plasma membrane. This study provides evidence that most of the beta APP secretase in H4 cells is intracellular, and confirms that the resulting C-terminal fragment is delivered to lysosomes immediately after cleavage. These results are discussed with regard to the possibility that mature full-length beta APP escapes secretase cleavage and is delivered directly from the TGN to the lysosome without passing through the plasma membrane. Either pathway will result in the generation of amyloidogenic fragments. PMID- 8240236 TI - Hepoxilin A3 inhibits the rise in free intracellular calcium evoked by formyl methionyl-leucyl-phenylalanine, platelet-activating factor and leukotriene B4. AB - We have previously shown that the hepoxilins are capable of increasing the intracellular free concentration of calcium ([Ca2+]i) in human neutrophils through a pertussis toxin-sensitive, extracellular calcium-independent pathway involving the mobilization of calcium from internal stores. A subsequent hepoxilin-induced and extracellular calcium-dependent influx of calcium is observed. In an effort to investigate further the role of these compounds in the human neutrophil, we investigated their potential effects on the action of known agonists such as formyl-methionine-leucine-phenylalanine (fMLP), platelet activating factor (PAF) and leukotriene B4 (LTB4) on the mobilization of calcium. Hepoxilis dose-dependently inhibited the increases in [Ca2+]i induced by fMLP, PAF and LTB4. The hepoxilin concentration required for inhibition was around 100 ng/ml (3 x 10(-7) M). This concentration of hepoxilin did not cause any measurable change in [Ca2+]i. The extent of inhibition of the agonist-evoked rise in [Ca2+]i by hepoxilins was proportional to the increase in the calcium response evoked by hepoxilin beyond its threshold concentration. Additional experiments were carried out to investigate the mechanism for the hepoxilin effect. Using calcium-free medium and in the presence of sufficient amounts of thapsigargin (200 ng/ml) to maximally block the calcium pump (thereby achieving a constant rate of calcium leakage from stores), hepoxilin A3 increased further this rate of calcium leakage, indicating that hepoxilin acts by rapidly draining calcium from stores. Its potential (additional) thapsigargin-like action in blocking the pump, however, cannot be ruled out by these experiments. These observations suggest that the hepoxilins may serve an important negative regulatory function in the agonist-induced mobilization of calcium in these cells by depleting calcium stores. PMID- 8240237 TI - Ca(2+)-calmodulin-dependent modification of smooth-muscle myosin light-chain kinase leading to its co-operative activation by calmodulin. AB - It has recently been shown that at relatively high molar ratios of myosin light chain kinase (MLCKase) to calmodulin (CM) almost complete inhibition of the kinase activity occurs [Sobieszek (1991) J. Mol. Biol. 220, 947-957]. This inhibition resulted in a highly co-operative activation of MLCKase by CM, whereas the opposite activation (of CM by kinase) was hyperbolic, as expected (unco operative). This difference in activation was observed only for kinase preparations preincubated with sub-stoichiometric amounts of CM, and only when micromolar concentrations of Ca2+ were present. The inhibitory effect was variable and depended not only on the concentration ratio of kinase to CM but also on the MLCKase preparation. For most of the preparations full inhibition required 5-15 min of preincubation at 25 degrees C and a 3-6-fold molar excess of kinase over CM. The inhibition was reversible, since full activity could be obtained after saturation of the kinase by additional CM. The inhibitory effect did not require ATP (excluding phosphorylation-type modifications of the kinase), and dephosphorylation of the kinase was not involved, since inhibition of an endogenous MLCK phosphatase by microcystin-LR did not decrease the inhibitory effect. Since the co-operative activation by CM was observed for cross-linked MLCKase preparations enriched in kinase dimers, but was absent for the analogous preparations enriched in the oligomers, we concluded that Ca(2+)-CM-dependent changes in the oligomeric state of the kinase were responsible for the modification observed. The exact nature of these modifications remains to be established. PMID- 8240238 TI - Purification and characterization of a phytase (myo-inositol-hexakisphosphate phosphohydrolase) accumulated in maize (Zea mays) seedlings during germination. AB - Phytase (myo-inositol-hexakisphosphate phosphohydrolase, EC 3.1.3.8) has been purified from 5-7-day-old maize (Zea mays) seedlings, using a four-step purification procedure. The native protein has a molecular mass of about 76 kDa and is built up from two 38 kDa subunits. The pH and temperature optima of the purified enzyme were respectively 4.8 and 55 degrees C. The apparent Km for phytate was estimated to be 117 microM. Like other acidic phytases, the maize seedling enzyme exhibited a broad affinity for various phosphorylated substrates and especially for penta- and tri-phosphate esters of myo-inositol. The amino acid composition of the h.p.l.c.-purified protein indicated a high hydrophobicity (44% non-polar amino acids). Rabbit antibodies were produced in response to maize seedling phytase. Western-blot analyses clearly demonstrate that the increase of phytase activity observed during the first 7 days of germination corresponded to an accumulation of the protein in maize seedlings. Phytase accumulated essentially in the shoots (mesocotyl plus coleoptiles. PMID- 8240239 TI - Non-proteoglycan forms of biglycan increase with age in human articular cartilage. AB - Polyclonal anti-peptide antibodies were raised to the C-terminal regions of human biglycan and decorin. These antibodies were used in immunoblotting to study structural variations with age in the proteoglycan core proteins present in extracts of human articular cartilage and intervertebral disc. Three forms of the biglycan core protein were identified. The largest form was detected only after chondroitinase treatment and represents the proteoglycan form of the molecule from which the glycosaminoglycan chains have been removed. However, chondroitinase treatment did not alter the electrophoretic mobility of the two smaller proteins, which appear to represent non-proteoglycan forms of the molecule, resulting either from a failure to substitute the intact proteoglycan core protein with glycosaminoglycan chains during its synthesis or from proteolytic processing of the intact proteoglycan causing removal of the N terminal region bearing the glycosaminoglycan chains. The non-proteoglycan forms constituted a minor proportion of biglycan in the newborn, but were the major components in the adult. A similar trend was seen in both articular cartilage and intervertebral disc. In comparison, decorin appears to exist predominantly as a proteoglycan at all ages, with two core protein sizes being present after chondroitinase treatment. Non-proteoglycan forms were detected in the adult, but they were always a minor constituent. PMID- 8240240 TI - The cytoplasmic domain of C-CAM is required for C-CAM-mediated adhesion function: studies of a C-CAM transcript containing an unspliced intron. AB - Cell-CAM105 (also named C-CAM) is a cell surface glycoprotein involved in intercellular adhesion of rat hepatocytes. It has four extracellular immunoglobulin (Ig) domains, a transmembrane domain and a cytoplasmic domain and therefore is a member of the Ig supergene family. We have characterized multiple cDNAs of the C-CAM genes in rat intestine. Sequence analyses showed that rat intestine contained not only the previously reported L-form and S-form C-CAMs (renamed C-CAM1 and C-CAM2 respectively) but also a new isoform, C-CAM3. The C CAM3 transcript codes for a polypeptide with a truncated C-terminus that lacks 65 amino acids from the previously reported C-CAM1 cytoplasmic domain. Unlike C CAM1, C-CAM3 did not mediate cell adhesion when expressed in insect cells using the baculoviral expression system. Thus the extra 65 amino acids in the cytoplasmic domain of C-CAM1 are important for adhesion phenotype when expressed in insect cells. Although C-CAM1 and C-CAM2 are encoded by different genes, sequence analysis suggests that C-CAM3 is probably derived from alternative splicing of the C-CAM1 gene. To examine this possibility, we have determined the exon organization of the C-CAM1 gene. C-CAM3 differed from C-CAM1 by the presence of a single unspliced intron which contained a stop codon immediately after the regular splice junction. As a result, translation of C-CAM3 terminates at the point where C-CAM1 and C-CAM3 sequences diverge. To investigate the expression of C-CAM1, C-CAM2 and C-CAM3 in different tissues, we used an RNAase-protection assay to simultaneously assess the levels of expression of these transcripts. Using total RNA prepared from various tissues, we showed that expression of C CAM3 was tissue-specific, and the C-CAM3 transcript accounted for about 25% of the transcripts derived from the C-CAM1 gene. However, further analysis revealed that C-CAM3 transcript was not present in cytosolic RNA, rather it was enriched in nuclear RNA prepared from hepatocytes. Although C-CAM3 cDNA contains the polyadenylation signal and is polyadenylated, these results indicate that C-CAM3 is probably an incomplete spliced product of C-CAM1 gene. PMID- 8240241 TI - Biochemical characterization of hamster oviductin as a sulphated zona pellucida binding glycoprotein. AB - Oviductins are a family of glycoproteins, synthesized and released by oviductal secretory cells, which bind to the zona pellucida of the oocyte after ovulation. Hamster oviductin migrates as diffuse species of 160-350 kDa during SDS/PAGE under reducing as well as non-reducing conditions. In this report, we describe the one-step purification of hamster oviductin using either immuno- or lectin affinity chromatography. Probing with specific lectins showed that the glycoprotein contains terminal alpha-D-GalNAc, and either terminal alpha-D-NeuAc or non-terminal beta-D-(GlcNAc)2 residues, but fails to react with concanavalin A and Ulex Europeus A-1 lectins which are specific for branched alpha-D-mannose and alpha-L-fucose moieties respectively. Intraovarian oocytes do not contain this glycoprotein and we demonstrate here that the immunoaffinity-purified oviductin readily binds to their zonae pellucidae in vitro, thus mimicking the in vivo phenomenon. Two major immunologically related forms of hamster oviductin (named alpha and beta) were characterized using one- and two-dimensional gel electrophoresis. The alpha-form (160-210 kDa) has an acidic pI of 3.5-4.5 and the beta-form (approx. 210-350 kDa) is localized at the cathodic site in the isoelectric focusing dimension; in between these two major forms lies a smear of minor-charge isomers. Peptide mapping of both major forms with papain and Staphylococcus aureus V8 protease yielded fragments of identical size. Moreover, the two forms share the same N-terminal sequence which display no significant homology with other reported proteins. Treatment with trifluoromethanesulphonic acid showed that a protein with the size and pI of the alpha-form can be generated from the beta-form. Both the alpha- and beta-forms are sulphated on O linked oligosaccharide side chains but are not phosphorylated. Collectively, these results suggest that the hamster oviductin polymorphism observed in two dimensional PAGE is a consequence of different glycosylation patterns and not the polypeptide chain itself. Hamster oviductin is mostly O-glycosylated and contains a few N-linked oligosaccharide side chains (approx. 10 kDa). We propose that hamster oviductin is a mucin-type glycoprotein which might act as a protective secretion influencing the first steps of the reproductive process necessary for the normal triggering of fertilization and early embryonic development. PMID- 8240242 TI - A membrane-bound form of glutamate dehydrogenase possesses an ATP-dependent high affinity microtubule-binding activity. AB - We previously identified a 50 kDa membrane protein which bound to in vitro assembled microtubules [Mithieux and Rousset (1989) J. Biol. Chem. 264, 4664 4668]. This protein exhibited the expected properties for mediating the ATP dependent association of vesicles with microtubules [Mithieux, Audebet and Rousset (1988) Biochim. Biophys. Acta 969, 121-130]. The 50 kDa membrane protein (MP50), initially extracted in very low amount from isolated pig thyroid lysosomes/endosomes, has now been purified from membrane preparations of crude vesicle fractions from pig liver and brain. MP50 was isolated from detergent solubilized membrane protein by affinity chromatography on immobilized ATP; 3-5 mg of MP50 was obtained from 100 g of liver tissue. Phase partitioning in Triton X-114 indicated that MP50 is a peripheral membrane protein. Radioiodinated liver MP50 bound to microtubules assembled in vitro. The binding was inhibited by ATP (Ki = 0.76 mM) and displaced by unlabelled liver or brain MP50. Equilibrium binding studies yielded KD values of 1.8 x 10(-7) M. By N-terminal amino acid sequence analysis, MP50 was identified as glutamate dehydrogenase (GDH), by comparison of V8 protease peptide maps of MP50 with purified liver GDH. Liver MP50 exhibited a low GDH activity; 4-5 units/mg compared with 18 and 34 units/mg for purified bovine and rat liver GDH respectively. Bovine and rat liver GDH yielded six spots from pI 5.7 to 7.2 when analysed by two-dimensional electrophoresis; in contrast, MP50 gave one main spot (corresponding to spot 2 of liver GDH) with a pI of approx. 6.5. Soluble liver GDH from commercial sources exhibited a very low or no microtubule-binding activity. In conclusion, we have found a membrane-bound form of GDH capable of specific and nucleotide-sensitive interaction with microtubules. Our data suggest that GDH isoproteins, the number of which has been undervalued up to now, could have cellular functions other than that of an enzyme. PMID- 8240243 TI - Expression of two different forms of cDNA for thromboxane synthase in insect cells and site-directed mutagenesis of a critical cysteine residue. AB - cDNA coding for human placental thromboxane synthase (EC 5.3.99.5) was amplified by PCR from a human placental cDNA library and sequenced. This cDNA and a shorter cDNA isolated from a human lung cDNA library with a deletion of 163 bp near the 3' end were expressed in Spodoptera frugiperda (Sf9) insect cells using a baculovirus expression system. The cDNA from human placenta was expressed as an active enzyme (60 kDa) with a specific activity higher than those reported from other cell types, whereas the shorter cDNA was expressed in an inactive form (52 kDa). The active recombinant enzyme appeared to be unglycosylated as the molecular mass and the enzyme activity were not altered in the presence of tunicamycin. Site-directed mutagenesis was performed to convert a cysteine at position 480 in thromboxane synthase to a serine. This cysteine is found to be highly conserved in related cytochrome P-450 enzymes. The mutant enzyme was found to be inactive, although Western blot, immunoprecipitation and SDS/PAGE analysis indicated that the mutant enzyme was expressed at a level comparable with the wild-type enzyme. These results suggest that Cys-480 is essential for the enzyme catalytic activity and that the short-form cDNA may be a non-functional transcript. PMID- 8240244 TI - Characterization of a chelator-resistant proteinase from Thermus strain Rt4A2. AB - The Thermus isolate Rt4A2 was found to produce an extracellular chelator resistant proteinase. The proteinase was purified to homogeneity by (NH4)2SO4 precipitation, cation-exchange chromatography, gel-filtration chromatography, and weak anion-exchange chromatography. The Rt4A2 proteinase was found to have properties typical of an alkaline serine proteinase. It had a pH optimum of 9.0 and was specifically inhibited by phenylmethanesulphonyl fluoride. Its isoelectric point was greater than 10.25. Its molecular-mass was 31.6 kDa as determined by SDS/PAGE. N-terminal sequencing has shown it to have high sequence similarity with other serine proteinases from Thermus species. The proteinase hydrolysed a number of substrates including fibrin, casein, haemoglobin, collagen, albumin and the synthetic chromogenic peptide substrate Suc-Ala-Ala-Pro Phe-NH-Np. The specific activity of the purified proteinase using azocasein as substrate was 313 units/mg. Substrate inhibition was observed above an azocasein concentration of 0.05% (w/v). Esterase activity was directed mainly towards those substrates containing the aliphatic or aromatic residues of alanine, glycine, tryptophan, tyrosine and phenylalanine. Thermostability half-lives of greater than 7 days at 70 degrees C, 43 h at 80 degrees C and 90 min at 90 degrees C were found in the presence of 5 mM CaCl2. At 90 degrees C increasing the CaCl2 concentration 100-fold (0.5 mM to 50 mM) caused a 4.3-fold increase in the half life of the enzyme from 30 to 130 min. Half-lives of 19.4 min at 100 degrees C and 4.4 min at 105 degrees C were found in the presence of 50 mM CaCl2. The metal chelators EGTA and EDTA reduced the stability at higher temperatures but had no effect on the activity of the proteinase. Activity was not stimulated by common metal activators such as Ca2+, Mg2+ and Zn2+. PMID- 8240245 TI - Isolation of highly purified, functional endosomes from toad urinary bladder. AB - Endosomes are difficult to isolate as they share size and density properties with much more abundant cellular organelles such as mitochondria. In cultured cell lines the tandem use of charge-dependent isolation techniques and differential centrifugation is necessary to isolate endosomes. Endosomal populations of the toad urinary bladder are of special interest because they are thought to contain a water channel. Understanding of the molecular structure of the water channel has been constrained, as there is currently no practical method to isolate functional water-channel-containing vesicles. This study reports the tandem use of charge-dependent techniques and centrifugation to isolate populations of endosomes from the toad urinary bladder. To purify water-channel-containing vesicles aqueous two-phase partition was utilized to fractionate a preparation partially purified by differential centrifugation. Populations of endosomes were analysed by small-particle flow cytometry techniques. A 5-fold enrichment in endosomes, achieved with aqueous two-phase partition, allowed us to identify two populations of endosomes of diverse size in a toad bladder endosomal fraction. Preenrichment also improved the efficiency of flow cytometry sorting, allowing isolation of the two endosomal populations in sufficient quantities for secondary analysis. A population of larger endosomes had very high water permeability, indicating the presence of water channels. The two populations had different SDS/PAGE fingerprints. Electron micrographs of the flow-sorted material shows a uniform population of membrane vesicles devoid of mitochondria and other identifiable cellular organelles. Hence, aqueous two-phase partition and flow cytometry allow identification of two populations of endosomes in the toad urinary bladder which have diverse structural and functional properties. Isolation of functional water-channel-containing vesicles allows co-localization of water-channel function with candidate water-channel proteins. PMID- 8240246 TI - Optimization of molecular design in the evolution of metabolism: the glycogen molecule. AB - The animal glycogen molecule has to be designed in accordance with its metabolic function as a very effective fuel store allowing quick release of large amounts of glucose. In addition, the design should account for a high capacity of glucose storage in the least possible space. We have studied the optimization of these variables by means of a mathematical model of the glycogen molecule. Our results demonstrate that the structure is optimized to maximize (a) the total glucose stored in the smallest possible volume, (b) the proportion of it that can be directly released by phosphorylase before any debranching occurs, and (c) the number of non-reducing ends (points of attack for phosphorylase), which maximizes the speed of fuel release. The optimization of these four variables is achieved with appropriate values for two key parameters in glycogen design: the degree of branching and the length of the chains. The optimal values of these two parameters are precisely those found in cellular glycogen. PMID- 8240247 TI - The role of cysteine residues 129 and 329 in Escherichia coli K1 CMP-NeuAc synthase. AB - N-Acetylneuraminic acid cytidyltransferase (CMP-NeuAc synthase) of Escherichia coli K1 is sensitive to mercurials and has cysteine residues only at positions 129 and 329. The role of these residues in the catalytic activity and structure of the protein has been investigated by site-directed mutagenesis and chemical modification. The enzyme is inactivated by the thiol-specific reagent dithiodipyridine. Inactivation by this reagent is decreased in the presence of the nucleotide substrate CTP, suggesting that a thiol residue is at or near the active site. Site-directed mutagenesis of either residue Cys-129 to serine or Cys 329 to selected amino acids has minor effects on the specific activity of the enzyme, suggesting that cysteine is not essential for catalysis and that a disulphide bond is not an essential structural component. The limited reactivity of the enzyme to other thiol-blocking reagents suggests that its cysteine residues are partially exposed. The accessibility and role of the cysteine residues in enzyme structure were investigated by fluorescence, c.d. and denaturation studies of wild-type and mutant enzymes. The mutation of Cys-129 to serine makes the enzyme more sensitive to heat and chemical denaturation, but does not cause gross changes in the protein structure as judged by the c.d. spectrum. The mutant containing Ser-129 instead of Cys-129 had a complex denaturation pathway similar to that of wild-type E. coli K1 CMP-NeuAc synthase consisting of several partially denatured states. Cys-329 reacts more readily with N-[14C]ethylmaleimide when the enzyme is in a heat-induced relaxed state. Cys-129 is less reactive and is probably a buried residue. PMID- 8240248 TI - Dissociation, unfolding and refolding trials of pig kidney 3,4 dihydroxyphenylalanine (dopa) decarboxylase. AB - The effect of guanidinium chloride (GuCl) on enzyme activity, hydrodynamic volume, circular dichroism, and fluorescence of 3,4-dihydroxyphenylalanine (Dopa) decarboxylase from pig kidney (pkDDC) was studied under equilibrium conditions. Unfolding proceeds in at least three stages. The first transition, occurring between 0 and 1 M GuCl, gives rise to a dimeric inactive species which has lost pyridoxal 5'-phosphate (PLP), and has a high tendency to aggregate, but retains almost all of the native spectroscopic characteristics. The second equilibrium transition, between 1 and 2.2 M GuCl, involves dimer dissociation, with some loss of tertiary and secondary structure. Additionally, gross conformational changes at or near the PLP microenvironment were detected by fluorescence of NaBH4 reduced enzyme. The third step, presumably representing complete unfolding of pkDDC, appears to be complete at 4.5 M GuCl, as indicated by the lack of further substantial changes in any of the signals being studied. Attempts at refolding resulted in the findings that: (1) partial reactivation is observed only starting from enzyme denatured at concentrations below 1.5 M GuCl, and (2) starting from completely denatured protein, the refolding process is apparently reversible down to concentrations of approx. 2 M GuCl. Taken together, this would seem to indicate that the monomer-dimer transition is impaired under the experimental conditions tested. A plausible model is presented for the unfolding/refolding of pkDDC. PMID- 8240249 TI - Expression in Escherichia coli of the flavin and the haem domains of Hansenula anomala flavocytochrome b2 (flavodehydrogenase and b2 core) and characterization of the recombinant proteins. AB - The flavin and haem domains of Hansenula anomala flavocytochrome b2 have been independently expressed in Escherichia coli. The flavin domain activity, studied only in the total cellular extract, owing to its instability, has characteristics very similar to those of the flavin domain obtained by proteolysis. The haem domain (r-core) has been purified to homogeneity and characterized in detail from spectroscopic and functional points of view. Spectral differences with respect to the domain produced by proteolysis (p-core) were found using resonance Raman and c.d. spectroscopy and have been interpreted in terms of changes in haem-protein interactions. However, this structural difference is functionally silent, since the r-core is able to reduce cytochrome c with the same efficiency as the proteolytic domain. PMID- 8240250 TI - Modification of 1 alpha,25-dihydroxyvitamin D3 metabolism by introduction of 26,26,26,27,27,27-hexafluoro atoms in human promyelocytic leukaemia (HL-60) cells: isolation and identification of a novel bioactive metabolite, 26,26,26,27,27,27-hexafluoro-1 alpha,23(S),25-trihydroxyvitamin D3. AB - To study whether the introduction of fluoro atoms into C-26 and C-27 positions on the 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3] molecule could affect metabolism in human promyelocytic leukaemia (HL-60) cells, we compared the metabolism of 26,26,26,27,27,27-hexafluoro-1 alpha,25-dihydroxyvitamin D3 [26,27 F6-1 alpha,25(OH)2D3] and 1 alpha,25(OH)2D3 in HL-60 cells. 26,27-F6-1 alpha,25(OH)2D3 was mainly converted into a new bioactive metabolite, 26,26,26,27,27,27-hexafluoro-1 alpha,23(S),25- trihydroxyvitamin D3 [26,27-F6-1 alpha,23(S),25(OH)3D3], but not into 26,26,26,27,27,27-hexafluoro-1 alpha,24(R),25-trihydroxyvitamin D3 [26,27-F6-1 alpha,24(R),25(OH)3D3] in HL-60 cells. 26,27-F6-1 alpha,23(S),25(OH)3D3 was identified by combinations of h.p.l.c., u.v. spectroscopy and g.c.-mass spectrometry. Evidence is presented that 26,27-F6-1 alpha,25(OH)2D2 was metabolized to 26,27-F6-1 alpha,23(S),25(OH)3D3 by C-23 hydroxylation as a first step of the metabolism, and the 23-hydroxylated bioactive metabolite was accumulated in the cells, whereas 1 alpha,25(OH)2D3 was initially deactivated and metabolized to 1 alpha,24(R),25(OH)3D3 by C-24 hydroxylation through a side-chain oxidation pathway resulting in C23-C24 cleavage, yielding 24,25,26,27-tetranor-1 alpha,23(OH)2D3 in HL-60 cells. These results show that 26,27-F6-1 alpha,25(OH)2D3 and 1 alpha,25(OH)2D3 are metabolized by different metabolic pathways in HL-60 cells. PMID- 8240251 TI - Isoprenoid biosynthesis in bacteria: a novel pathway for the early steps leading to isopentenyl diphosphate. AB - Incorporation of 13C-labelled glucose, acetate, pyruvate or erythrose allowed the determination of the origin of the carbon atoms of triterpenoids of the hopane series and/or of the ubiquinones from several bacteria (Zymomonas mobilis, Methylobacterium fujisawaense, Escherichia coli and Alicyclobacillus acidoterrestris) confirmed our earlier results obtained by incorporation of 13C labelled acetate into the hopanoids of other bacteria and led to the identification of a novel biosynthetic route for the early steps of isoprenoid biosynthesis. The C5 framework of isoprenic units results most probably (i) from the condensation of a C2 unit derived from pyruvate decarboxylation (e.g. thiamine-activated acetaldehyde) on the C-2 carbonyl group of a triose phosphate derivative issued probably from dihydroxyacetone phosphate and not from pyruvate and (ii) from a transposition step. Although this hypothetical biosynthetic pathway resembles that of L-valine biosynthesis, this amino acid or its C5 precursors could be excluded as intermediates in the formation of isoprenic units. PMID- 8240252 TI - Econazole inhibits thapsigargin-induced platelet calcium influx by mechanisms other than cytochrome P-450 inhibition. AB - Cytochrome P-450 has been suggested as a mediator of the signal between depleted platelet calcium stores and an increase in plasma membrane permeability to calcium which follows depletion of the stores. This hypothesis is based on the observations that inhibitors of cytochrome P-450, such as the imidazole antifungal agents, also inhibit influx of a calcium surrogate (manganese) into calcium-depleted platelets. We tested the effects of econazole and of a cytochrome P-450 inhibitor, carbon monoxide (CO), on thapsigargin (TG)-induced platelet 45Ca2+ influx. TG specifically depletes internal calcium stores and activates store-regulated calcium influx. Econazole blocked 45Ca2+ influx when it was added before TG (IC50 11 microM). Econazole at a concentration (20 microM) that inhibited 83% of TG-induced calcium influx was not inhibitory to TG-induced calcium efflux from 45Ca(2+)-loaded platelets, and did not affect calcium fluxes in resting platelets. This econazole concentration was also inhibitory to calcium influx even when it was added after the stores had been calcium-depleted by EGTA and TG for 15 min and the signal to increase calcium influx had already been generated. Inhibition of cytochrome P-450 with CO bubbled through platelet suspensions did not change calcium influx in resting cells and potentiated TG induced calcium influx (160% of control calcium accumulation at 20 min). This effect appeared to be concentration-dependent, such that a 5 min exposure to CO produced a greater influx potentiation than a 3 min exposure. These observations indicate that (1) cytochrome P-450 does not mediate store-regulated calcium influx, and (2) econazole probably inhibits store-regulated calcium influx by an alternative mechanism, such as interaction with plasma membrane calcium channels. PMID- 8240253 TI - The protein phosphatases responsible for dephosphorylation of hormone-sensitive lipase in isolated rat adipocytes. AB - The levels of the cytosolic serine/threonine protein phosphatases (PP) in rat adipocyte extracts have been determined, by using both reference substrates and hormone-sensitive lipase (HSL) as substrates. Adipocytes contain significant levels of both PP1 and 2A (1.6 and 2.0 m-units/ml of packed cells respectively), with lower levels of PP2C and virtually no PP2B activity. PP2A and 2C exhibit similar degrees of activity against HSL phosphorylated at site 1, together accounting for 92% of the total. In contrast, site 2 is dephosphorylated predominantly by PP2A (over 50% of total activity), whereas PP1 and PP2C contribute approx. 20% and 30% respectively to the total phosphatase activity against that site. Total phosphatase activity in the adipocyte extracts was 2-3 fold higher against site 2 than against site 1. The possible significance of these findings to the regulation of HSL activity in adipose tissue in vivo is discussed. PMID- 8240254 TI - Assignment of laminin heavy chains using the lectin Ricinus communis agglutinin 1. AB - Using high-resolution PAGE and Western-blotting techniques the lectin Ricinus communis agglutinin-1 (RCA-1) was tested for its ability to recognize laminin subunits from the mouse Engelbreth-Holm-Swarm (EHS) tumour and from bovine cardiac and skeletal muscle. Biotinylated RCA-1 recognized both the A and B chains of purified EHS-tumour laminin with a sensitivity comparable to anti-(EHS laminin) antibodies. In cardiac and skeletal muscle RCA-1 also recognized the B chains of laminin, together with a approximately 330 kDa RCA-1-binding glycoprotein that was undetectable in smooth muscle. This glycoprotein was not recognized by antibodies raised to laminin from the EHS tumour. Purification of the 330 kDa binding glycoprotein from skeletal muscle, using ion-exchange and lectin-affinity chromatography, revealed that in its native form, this glycoprotein is disulphide-bonded to the B chains of laminin. The demonstrated properties of the approximately 330 kDa RCA-1-binding glycoprotein are identical to those reported for the variant M chain of merosin which is known to replace the A chain in laminin from the extrasynaptic regions of skeletal muscle. These results establish that biotinylated RCA-1 can recognize A-, B- and M-chain subunits of laminin isoforms, and that, when used in conjunction with other techniques, they provide a useful method for the assignment of laminin heavy chains. PMID- 8240255 TI - Induction of pterin synthesis is not required for cytokine-stimulated tryptophan metabolism. AB - Activation of the immune system which occurs in inflammatory disease leads to parallel increases in pterin synthesis and increased production of neuroactive L tryptophan metabolites. Several model systems were studied to determine whether pterins, which are cofactors for hydroxylation reactions, could be required in the oxidative kynurenine pathway of L-tryptophan degradation. Treatment of mice with interferon-gamma increased L-tryptophan metabolism without any corresponding change in tissue biopterin concentrations. Cytokine-treated human fibroblasts, macrophages and glioblastoma cells all showed increases in kynurenine production, which were completely independent of pterin synthesis. When pterin synthesis de novo was blocked, either by an inhibitor of GTP cyclohydrolase or because of a genetic deficiency of one of the enzymes of the pathway of pterin biosynthesis, cytokine-stimulated increases in tryptophan metabolism were unaffected. Furthermore, increasing intracellular tetrahydrobiopterin concentrations by treating cells with sepia-pterin also had no effect on markers of tryptophan metabolism. Therefore, both normal and cytokine-stimulated L-tryptophan metabolism appears to be completely independent of pterin biosynthesis. PMID- 8240256 TI - Histone H4 stimulates glucose transport activity in rat skeletal muscle. AB - We investigated the effects of purified histone H4 on glucose transport activity in rat soleus and flexor digitorum brevis muscles. Histone H4, at concentrations up to 11.8 microM, increased 2-deoxyglucose (2-DG) uptake in a dose-dependent fashion. However, at concentrations higher than 11.8 microM, H4 caused a decrease in 2-DG uptake from the maximum, suggesting a secondary inhibitory action of this compound. The maximal effect of H4 on 2-DG uptake was not additive to the maximal effect of insulin. Moreover, 2-DG uptake in the presence of both H4 and insulin was significantly lower than the 2-DG uptake in the presence of insulin alone. The maximal effect of H4 on stimulation of 2-DG uptake was neither additive nor inhibitory to the maximal effects of the intracellularly acting insulin mimetics sodium vanadate or H2O2. It was, on the other hand, additive to the maximal effects of muscle contractions. Also, in contrast with the effects of H4 on insulin-stimulated 2-DG uptake, H4 did not inhibit insulin-like growth factor-I (IGF-I)-stimulated 2-DG uptake, as the maximal effects of H4 and IGF-I were additive. Scatchard analysis of the binding of 125I-insulin in the absence or presence of histone H4 revealed that H4 increased the specific binding of insulin without affecting receptor affinity. These data suggest that H4 interacts with the insulin, rather than the hypoxia/contraction, pathway for activation of glucose transport in muscle tissue, and that H4 acts either directly or indirectly to increase the number of insulin receptors at the surface of the muscle cell. This interaction does not appear to occur with the similar, although distinct, IGF-I receptor. These studies may provide additional insight into the complex signal-transduction systems of insulin action. PMID- 8240257 TI - Characterization of glycoinositol phospholipids in the amastigote stage of the protozoan parasite Leishmania major. AB - The major macromolecules on the surface of the parasitic protozoan Leishmania major appear to be down-regulated during transformation of the parasite from an insect-dwelling promastigote stage to an intracellular amastigote stage that invades mammalian macrophages. In contrast, the major parasite glycolipids, the glycoinositol phospholipids (GIPLs), are shown here to be expressed at near constant levels in both developmental stages. The structures of the GIPLs from tissue-derived amastigotes have been determined by h.p.l.c. analysis of the deaminated and reduced glycan head groups, and by chemical and enzymic sequencing. The deduced structures appear to form a complete biosynthetic series, ranging from Man alpha 1-4GlcN-phosphatidylinositol (PI) to Gal alpha 1-3Galf beta 1-3Man alpha 1-3Man alpha 1-4GlcN-PI (GIPL-2). A small proportion of GIPL-2 was further extended by addition of a Gal residue in either alpha 1-6 or beta 1-3 linkage. From g.c.-m.s. analysis and mild base treatment, all the GIPLs were shown to contain either alkylacylglycerol or lyso-alkylglycerol lipid moieties, where the alkyl chains were predominantly C18:0, with lower levels of C20:0, C22:0 and C24:0. L. major amastigotes also contained at least two PI-specific phospholipase C-resistant glycolipids which are absent from promastigotes. These neutral glycolipids were resistant to both mild acid and mild base hydrolysis, contained terminal beta-Gal residues and were not lost during extensive purification of amastigotes from host cell membranes. It is likely that these glycolipids are glycosphingolipids acquired from the mammalian host. The GIPL profile of L. major amastigotes is compared with the profiles found in L. major promastigotes and L. donovani amastigotes. PMID- 8240258 TI - The interaction of cytosolic components of neutrophil NADPH oxidase with phosphoinositides. AB - The superoxide-generating NADPH oxidase of neutrophils can be activated in a cell free system consisting of cell membranes, cytosol and an activating detergent (e.g. arachidonate or SDS). It has previously been reported [Aviram and Sharabani (1989) Biochem. Biophys. Res. Commun. 161, 712-719] that a mixture of phosphoinositides (PPIs), as well as the individual inositol lipids, interfere with the activation process. In the present study it is shown that exposure of the cytosol to PPI results in a progressive (t1/2 = 30 s) loss of its oxidase supporting activity and that Mg2+ ions eliminate this inactivation. Neomycin, previously described as an inhibitor of cell-free activation, counteracted the effect of PPI and vice versa. Fractionation experiments implicated the p67-phox cytosolic component of the oxidase in the association with PPI. PPI blocked activity of recombinant p67-phox also and quenched the fluorescence intensity of its tryptophan residues. It is suggested that PPIs may mediate the interaction of the oxidase with the cytoskeleton and/or with the membrane. PMID- 8240259 TI - Expression of a rat neurotensin receptor in Escherichia coli. AB - With the goal of obtaining sufficient quantities of seven-helix G-protein-coupled receptors for structural analysis, we have studied the functional expression of a rat neurotensin receptor cDNA in Escherichia coli with and without a signal sequence and as a fusion with the gene coding for maltose-binding protein. The addition of an N-terminal signal peptide resulted in increased expression levels. In vitro translation at a high level revealed that the codon usage of the rat neurotensin receptor cDNA was not critical for overproduction. Expression of neurotensin receptor cDNA fused to the 3' end of the gene encoding maltose binding protein resulted in a 40-fold increase in neurotensin-binding sites. Binding of [3H]neurotensin to intact bacteria or E. coli membranes was saturable, with a dissociation constant, KD, of 0.23 nM (Bmax. = 450 sites/bacterium or 15 pmol/mg of crude membrane protein). The binding properties of all recombinant receptors presented in this study were similar and corresponded to those of the high-affinity binding sites in rat brain. For immunological detection and future purification of neurotensin receptor, a C-terminal pentahistidine/c-myc tail was introduced. Western-blot analysis revealed the association of neurotensin receptor with E. coli membranes. PMID- 8240260 TI - Protein catabolism in fibroblasts cultured from patients with mucolipidosis II and other lysosomal disorders. AB - Protein catabolism in fibroblasts cultured from the skin of normal individuals and of patients with mucolipidosis II (I-cell disease) and several other lysosomal storage diseases was examined by metabolic labelling with [3H]leucine and following the fate of radioactive proteins in pulse-chase experiments. In mucolipidosis II cells, overall protein degradative rates were found to be distinctly lower than in normal control cells. To distinguish lysosomal from non lysosomal degradation, labelling experiments were carried out in the presence and absence of 10 mM NH4Cl, an inhibitor of lysosomal function. It was found that mucolipidosis II fibroblasts exhibited a markedly reduced rate of lysosomal protein degradation, whereas the rate of nonlysosomal degradation appeared normal. Serum and amino acid starvation led to a marked increase in lysosomal protein degradation in normal cells, but had only a minimal effect on that in mucolipidosis II fibroblasts. The specific activities of cathepsins B, H and L were profoundly diminished in all mucolipidosis II cell lines tested. Lysosomal protein degradation in a mucolipidosis III cell line was impaired to a similar degree as in mucolipidosis II cells, whereas it was decreased to a lesser extent in fibroblasts from patients with mucopolysaccharidoses I and VI, galactosialidosis and GM1-gangliosidosis. We conclude that fibroblasts from patients with mucolipidosis II and III have a severely compromised capacity for endogenous lysosomal protein degradation that appears to result from multiple cathepsin deficiency. This lysosomal defect is likely to have pathophysiological consequences. PMID- 8240261 TI - Direct activation of human neutrophil procollagenase by recombinant stromelysin. AB - Human neutrophil procollagenase was activated by incubation with recombinant active stromelysin. Activation was achieved by cleavage of the Gly78-Phe79 peptide bond at the end of the propeptide domain in a single-step activation mechanism. In addition, accelerated activation was achieved when N-terminally truncated, latent collagenase (with Phe49 as its N-terminal residue) was incubated with recombinant active stromelysin. Determination of the specific activity of recombinant-stromelysin-activated neutrophil collagenase with dinitrophenyl-octapeptide or type I collagen demonstrated the generation of high specific activity. The specific activity of stromelysin-activated enzyme was considerably higher than that of trypsin- or HgCl2-activated collagenase. Thus human neutrophil collagenase is superactivated, like the homologous fibroblast collagenase [Murphy, Cockett, Stephens, Smith and Docherty (1987) Biochem. J. 248, 265-268]. The occurrence of Phe79 at the N-terminus of the neutrophil collagenase seemed to be critical for superactivation, which is in agreement with data published by Suzuki, Enghild, Morodomi, Salvesen and Nagase [(1990) Biochemistry 29, 10261-10270] on fibroblast collagenase. PMID- 8240262 TI - Purification and characterization of the dissimilatory nitrite reductase from Alcaligenes xylosoxidans subsp. xylosoxidans (N.C.I.M.B. 11015): evidence for the presence of both type 1 and type 2 copper centres. AB - Dissimilatory nitrite reductase was isolated from extracts of Alcaligenes xylosoxidans subsp. xylosoxidans (N.C.I.M.B. 11015), after activation of crude extracts by the addition of copper(II) sulphate. The enzyme was purified by a combination of (NH4)2SO4 fractionation and cationic-exchange chromatography to 93% homogeneity as judged by SDS/PAGE. SDS/PAGE and spray m.s. showed that the enzyme had a subunit M(r) of 36.5 kDa. The copper content was 3.5 +/- 0.8 Cu atoms/trimer of M(r) 109,500. E.p.r. spectroscopy of nitrite reductase as isolated showed that both type 1 (g parallel = 2.208, A parallel = 6.3 mT) and type 2 (g parallel = 2.298, A parallel = 14.2 mT) Cu centres were present, in contrast with published data [Masuko, Iwasaki, Sakurai, Suzuki and Nakahara (1984) J. Biochem. (Tokyo) 96, 447-454], where only type 1 copper centres were reported. Our preparations had a specific activity of 150-300 mumol of NO2- reduced/min per mg of protein, 6-12-fold higher than reported previously. As isolated, the oxidized form of our preparations of the enzyme showed absorption maxima in the visible region at 460, 593 and 770 nm. The ratio of the absorption bands at 460 nm and 593 nm resulted in this protein having a strong blue colour, in contrast with the green colour of other purified copper-containing nitrite reductases. We conclude that, in contrast with previous reports, this 'blue' nitrite reductase requires both type 1 and type 2 copper centres for optimal activity. PMID- 8240263 TI - Inositol lipid signalling occurs in brush-border membranes during initiation of compensatory renal growth in the rat. AB - Using highly specific mass assays, concentrations of inositol lipids and 1,2 diacylglycerol (DAG) were determined in plasma membranes isolated from rat kidney cortex. Significantly higher concentrations of inositol lipids were determined in brush-border (BBM) than in basal-lateral (BLM) plasma membranes, although DAG concentrations were similar in both. After unilateral nephrectomy, a decrease in PtdIns(4,5)P2 and PtdIns4P, with a concomitant increase in DAG and translocation of protein kinase C (PKC), were observed in BBM but not in BLM isolated from the remaining kidney. On the other hand, stimulation of renal cortical slices with insulin-like growth factor II (IGF-II) or phenylephrine caused similar effects in BLM but not in BBM. Stimulation of phospholipase C activity with translocation of PKC only to BBM in one kidney was also induced by occlusion of blood flow through the contralateral kidney for 15 min. At 30 min after the occlusion was removed and reflow established, DAG concentration and the amount of PKC in BBM returned to control values. These results suggest that an early signal after unilateral nephrectomy is transmitted to cells through BBM and can be switched on and off by blood occlusion and reflow through the contralateral kidney, while hormonal signals caused by IGF-II and phenylephrine are transmitted to cells through BLM. PMID- 8240264 TI - Does phosphoglucoisomerase display anomeric specificity or selectivity towards alpha-D-glucose 6-phosphate? An assessment by two-dimensional phase-sensitive 31P exchange spectroscopy (EXSY) n.m.r. AB - The study by two-dimensional phase-sensitive 31P exchange spectroscopy (EXSY) n.m.r. of hexose 6-phosphates interconversion in the reaction catalysed by yeast phosphoglucoisomerase reveals that the enzyme displays anomeric selectivity, rather than specificity, towards alpha-D-glucose 6-phosphate. Indeed, beta-D glucose 6-phosphate participates for about 20% to the total and direct conversion of the aldohexose into oxohexose ester. PMID- 8240265 TI - Evidence that the glutamine-stimulated loss of nitrate reductase protein from the yeast Candida nitratophila is not the result of inducer exclusion. AB - Synthesis of nitrate reductase protein and increases in nitrate reductase activity occurred in cultures of the yeast Candida nitratophila when they were incubated in medium containing ammonium nitrate. Similar treatment with glutamine plus nitrate resulted in little increase in nitrate reductase activity, in cultures grown previously with reduced nitrogen compounds, and decreases in enzyme activity, in cultures adapted to nitrate. Labelling studies conducted in vivo revealed a rapid cessation of de novo nitrate reductase synthesis when glutamine was supplied to nitrate-adapted cultures in the presence of nitrate. Intracellular glutamine concentrations increased rapidly under these conditions and these cultures exhibited high glutamine: glutamate ratios. As nitrate was taken up in the presence of glutamine in these experiments, it is concluded that the glutamine-stimulated inhibition of nitrate reductase synthesis is a consequence of repression and rapid turnover of nitrate reductase mRNA and not inducer (nitrate) exclusion. PMID- 8240266 TI - A quantitative analysis of the control of glutamine catabolism in rat liver cells. Use of selective inhibitors. AB - 1. At a physiological concentration of glutamine (0.5 mM), 87% of the total transport across the plasma membrane of liver cells isolated from fed rats involved the Na(+)-dependent system N; this was substantially inhibited by L histidine. The residual Na(+)-independent component was attributed to system L on the basis of inhibition by 2-amino-2-norbornanecarboxylate and L-tryptophan. 2. Catabolism of glutamine by intact liver cells or by isolated mitochondria was inhibited by glutamate gamma-hydrazide with IC50 values of 13.7 +/- 3.5 microM and 22.6 +/- 3.8 microM respectively and a maximal inhibition of approx. 75%. The site of inhibition was identified as glutaminase; glutamate gamma-hydrazide inhibited this enzyme in cell-free extracts (IC50 37.8 +/- 7.7 microM) but had no activity against glutamate dehydrogenase or transport of glutamine, whether across mitochondrial or plasma membranes. 3. The major control site in cells from fed animals incubated with 0.5 mM L-glutamine was glutaminase (flux control coefficient 0.96). Appreciable control also resided in both plasma membrane transport systems, with coefficients of 0.51 for system N and -0.46 for system L, such that both interacted to provide a fine control of the intracellular concentration of the amino acid. Similar values were obtained by computer simulation based on theoretical determination of elasticities. 4. Previous controversy about the locus of regulation of hepatic glutamine metabolism is resolved by this distribution of control. PMID- 8240267 TI - Molecular cloning of a rat kappa opioid receptor reveals sequence similarities to the mu and delta opioid receptors. AB - By screening a rat brain cDNA library using a cloned mu opioid receptor cDNA as probe, a clone was identified that is very similar to both the mu and delta opioid receptor sequences. Transient expression of this clone in COS-7 cells showed that it encodes a kappa opioid receptor, designated KOR-1, which is capable of high-affinity binding to kappa-selective ligands. Treatment of transfected cell membranes with bremazocine, a kappa-selective agonist, resulted in a 53% decrease in adenylate cyclase activity, indicating that this kappa opioid receptor displays inhibitory coupling to adenylate cyclase. Thus, one member from each of the three opioid receptor types, mu, kappa and delta, has been molecularly cloned. Analysis of sequence similarities among these opioid receptors, as well as between opioid receptors and other G-protein-coupled receptors, revealed regions of sequence conservation that may underlie the ligand binding and functional specificities of opioid receptors. PMID- 8240268 TI - Molecular cloning and expression of a rat kappa opioid receptor. AB - At least three types of opioid receptors have been identified in the nervous system. In this paper we report molecular cloning and expression of a rat kappa opioid receptor. PCR was performed on double-stranded cDNA derived from poly(A)+ RNA of the rat striatum with primers similar to those of Libert and co-workers [Libert, Parmentier, Lefort, Dinsart, Van Sande, Maenhaut, Simons, Dumont and Vassart (1989) Science 244, 569-572]. One of the PCR products, which had 65% sequence similarity to the mouse delta opioid receptor, was used to screen a rat striatum cDNA library. Two positive clones were isolated and found to be identical. The clone had a 2.1-kb insert, which was termed RKOR-1. RKOR-1 has an open reading frame of 1140 bp and encodes a 380-amino-acid protein. Hydropathy analysis indicates that RKOR-1 has seven putative transmembrane domains with short intra- and extra-cellular loops. Membranes of Cos-7 cells transfected with RKOR-1 exhibited high specific binding for [3H]diprenorphine ([3H]DIP), a non selective opioid ligand. Naloxone inhibited [3H]DIP binding with stereospecificity. [3H]DIP binding was potently inhibited by selective kappa opioid ligands, with Ki values in the nanomolar or subnanomolar range, but much less potently inhibited by drugs selective for mu or delta receptors. Thus, RKOR 1 represents an opioid receptor with kappa characteristics. PMID- 8240269 TI - Stable correction of maple syrup urine disease in cells from a Mennonite patient by retroviral-mediated gene transfer. AB - We have successfully used retroviral gene transfer to correct the deficiency of the branched-chain alpha-oxo acid dehydrogenase complex in lymphoblasts from a homozygous Mennonite maple syrup urine disease (MSUD) patient. The mutation in Mennonites is a Tyr-393 to Asn substitution in the branched-chain alpha-oxo acid decarboxylase (E1)alpha subunit of the enzyme complex. This promotes improper assembly of mutant E1 alpha with E1 beta subunits, leading to degradation of both polypeptides. For transduction studies, a full-length human E1 alpha CDNA was inserted into the retroviral vector LXSN to produce the recombinant LSN-E1 alpha. High-titre [6 x 10(5) colony-forming units/ml] amphotropic retroviral preparations free of helper viruses were obtained by co-cultivation of infected GP+E86 with PA317 cells. Transduction of MSUD lymphoblasts from the Mennonite patient with LSN-E1 alpha viruses restored the decarboxylation of alpha-oxo[1 14C]isovalerate to the normal level. The normal decarboxylation activity in transduced MSUD cells remained stable without G418 selection during the 14 weeks studied. Southern-blot analysis indicated that the recombinant E1 alpha cDNA was integrated into the host genome. Northern and Western blotting showed that both the normal E1 alpha mRNA and the subunit were properly expressed in transduced MSUD cells. However, the level of E1 beta subunits is lower than that of normal cells, suggesting competition of the recombinant E1 alpha with the mutant form for assembly with E1 beta. The results provide a paradigm for the development of somatic gene therapy for disorders involving mitochondrial multienzyme complexes. PMID- 8240270 TI - Retinoic acid regulates ornithine decarboxylase gene expression at the transcriptional level. AB - Retinoic acid (RA) is important for normal mammalian development and growth. Ornithine decarboxylase (ODC) is the first and rate-limiting enzyme in the biosynthesis of the polyamines, and we have previously shown that ODC mRNA levels are suppressed by RA in human skin cells. Using HeLa cells, we now show that treatment with 0.5 microM RA for 24 h suppresses endogenous ODC mRNA levels and the expression of a transfected ODC/chloramphenicol acetyltransferase plasmid (Kpn-ODCCAT), containing sequences from -1450 to +810 of the human ODC gene. Co transfection with either the alpha-RA receptor (alpha-RAR) or a chimeric alpha RA/oestrogen receptor (alpha-RAER) followed by treatment with the cognate hormone suppresses expression of Kpn-ODCCAT and Not-ODCCAT, which contains sequences from -250 to +514. Liganded alpha-RAR suppresses the activity of Kpn-ODCCAT more markedly than does liganded alpha-RAER (98% and 80% suppression, respectively), whereas both receptors have very similar effects on Not-ODCCAT expression (73% and 67% suppression, respectively). The unliganded alpha-RAR suppresses Kpn ODCCAT by 76%, whereas unliganded alpha-RAER has no significant effect. These data show that RA regulates ODC-gene expression at the transcriptional level, and that alpha-RAR, but not alpha-RAER, can confer full hormonal responsiveness. This suggests that the activating function present in the alpha-RAR ligand-binding domain is required for full transcriptional regulation. PMID- 8240271 TI - Glucosamine-6-phosphate deaminase from Escherichia coli has a trimer of dimers structure with three intersubunit disulphides. AB - Glucosamine-6-phosphate deaminase is an oligomeric protein composed of six identical 29.7 kDa subunits. Each subunit has four cysteine residues located at positions 118, 219, 228 and 239. We have previously shown that Cys-118 and Cys 239 form a pair of vicinal thiols, the reactivity of which changes with the allosteric transition. The site-directed mutations Cys-->Ser corresponding to the other two cysteine residues have been constructed, as well as some selected multiple mutations involving the four cysteines. Thiol and disulphide measurements on the wild-type and mutant enzymes indicate that thiols from Cys 219 are oxidized and form interchain disulphide bonds. The disulphide-linked dimer was demonstrated by SDS/PAGE. This result is consistent with preliminary crystallographic data and thermal denaturation studies, and strongly suggests that glucosamine-6-phosphate deaminase is a trimer of disulphide-linked dimers. The mutant forms of the deaminase lacking the interchain disulphide bond or the thiol at Cys-228 are both stable hexamers showing the same sensitivity to urea denaturation as the wild-type protein. Furthermore, these Cys-->Ser mutants display the same kinetics and allosteric properties as those already described for the wild-type enzyme. PMID- 8240272 TI - Post-translational processing of chromogranin A: differential distribution of phosphorylated variants of pancreastatin and fragments 248-313 and 297-313 in bovine pancreas and ileum. AB - Chromogranin A is a secretory protein expressed widely in neuroendocrine cells. It is known to be phosphorylated but the precise sites of phosphorylation are not known. We have isolated, from bovine pancreas and ileum, chromogranin A fragments corresponding to a region giving rise to a biologically active product, pancreastatin. Phosphorylation patterns were determined by fast atom bombardment mass spectrometry and alkaline phosphatase digestion followed by ion-exchange chromatography and radioimmunoassay. In the pancreas, there were unmodified, mono and di-phosphorylated forms of the fragment chromogranin A(248-313) with Arg and Glu at positions 293 and 301 respectively; in addition, there were small amounts of monophosphorylated peptide with an alternative primary sequence of His and Lys at 293 and 301 respectively. Two products of cleavage, pancreastatin and the fragment 297-313, were also found in unmodified and monophosphorylated forms. In the ileum, peptides with both alternative primary sequences were found, pancreastatin was absent, and phosphorylation was generally less than in the pancreas. Chromogranin A-derived peptides therefore exhibit tissue-specific patterns of phosphorylation and cleavage, and at least two phosphorylation sites occur in the region giving rise to a biologically active product. PMID- 8240273 TI - Attenuation of Gs alpha coupling efficiency in brown-adipose-tissue plasma membranes from cold-acclimated hamsters. AB - In order to localize site(s) of beta-adrenergic desensitization found in brown adipocytes from cold-acclimated animals, total brown-adipose-tissue homogenates (postnuclear supernatant) were obtained from control or cold-acclimated hamsters and were fractionated on discontinuous sucrose gradients. A low-density band (cytosolic proteins) and a high-density band (mitochondria) were obtained; in the middle fractions only low levels of protein were recovered. However, these fractions displayed a high level of specific [3H]ouabain binding, indicating that they represented fractions enriched in plasma membranes. The level of [3H]ouabain binding was significantly higher in plasma membranes from cold-acclimated animals, indicating an increased density of Na,K-ATPase units. The maximal activity of adenylate cyclase, as estimated with forskolin, was not changed by cold acclimation. However, the levels of cyclase activity observed after Gs protein-mediated activation (with guanosine 5'-[gamma-thio]triphosphate, isoprenaline, both of these, or fluoride) were decreased, indicating a decreased coupling efficiency. Notably, a significant decrease was observed in the functional activity of the Gs protein, as directly measured by estimation of the ability of cholate extracts of brown-fat plasma membranes to reconstitute Gs protein-mediated stimulation of adenylate cyclase in cyc- membranes. Further, a functionally significant decrease (to 72%) was observed in the ratio between the amount of functional Gs proteins and adenylate cyclase units. The total content of Gs alpha protein was decreased to the same extent as the coupling efficiency of the membranes, indicating that a lower content of functionally equivalent Gs alpha molecules could explain the decreased coupling. It could therefore be concluded that a decrease in Gs-protein-mediated coupling efficiency, owing to a decrease in the amount of Gs alpha, is at least one site of beta-adrenergic desensitization in cold-acclimated animals. This may, at least in part, explain that desensitization takes place despite the fact that the beta 3-adrenoceptor itself apparently lacks some of the sites known to be involved in the desensitization process in other beta-adrenergic receptors. PMID- 8240274 TI - CoA and fatty acyl-CoA derivatives mobilize calcium from a liver reticular pool. AB - The effect of CoA and fatty acyl-CoA esters on Ca2+ fluxes has been studied in isolated liver microsomes and in digitonin-permeabilized hepatocytes. When microsomes were loaded with increasing concentrations of Ca2+ (6-29 nmol/mg of protein), the extent to which CoA and palmitoyl-CoA released Ca2+ increased. At 23 nmol of Ca2+/mg of protein, half-maximal [CoA] and [palmitoyl-CoA] were 35 and 50 microM respectively. Under conditions of minimal Ca2+ loading, net release of Ca2+ was absent, but Ca2+ translocation from a CoA-sensitive to a CoA-insensitive pool took place. The effect of CoA required the presence of fatty acids, probably to form fatty acyl esters. In permeabilized hepatocytes, the pool(s) mobilized by CoA (or by palmitoyl-CoA) appeared to be different from that mobilized by Ins(1,4,5)P3. PMID- 8240275 TI - Changes in molar volume and heat capacity of actin upon polymerization. AB - We have used densimetry and microcalorimetry to measure the changes in molar volume and heat capacity of the actin molecule during Mg(2+)-induced polymerization. Molar volume is decreased by 720 ml/mol. This result is in contradiction with previous measurements by Ikkai and Ooi [(1966) Science 152, 1756-1757], and by Swezey and Somero [(1985) Biochemistry 24, 852-860]: both of these groups reported increases in actin volume during polymerization, of 391 ml/mol and 63 ml/mol respectively. We also observed a decrease in heat capacity of about 69.5 kJ.K-1.mol-1 during polymerization. This is in agreement with the concept of conformational fluctuation of proteins proposed by Lumry and Gregory [(1989) J.Mol. Liq. 42, 113-144]whereby either ligand binding by a protein or monomer-monomer interaction decreases the protein's conformational flexibility. PMID- 8240276 TI - Glucokinase activity in isolated islets from obese fa/fa Zucker rats. AB - Glucokinase (EC 2.7.1.2) activity of B-cells was measured in extracted pancreatic islets isolated from lean and obese fa/fa Zucker rats and maintained in primary culture overnight. Formation of [14C]glucose phosphoric esters from D-[U 14C]glucose was measured in the presence of unlabelled glucose from 0.05 to 0.50 mM for hexokinase (EC 2.7.1.1) activity, and 8.0-16.0 mM unlabelled glucose for glucokinase activity. Eadie-Hofstee analysis revealed that hexokinase kinetic parameters (Vmax and Km) for [14C]glucose phosphoric ester formation were similar in lean- and fa/fa-rat islets. For glucokinase, there was no difference in Vmax. between phenotypes. A non-significant tendency to increased sensitivity to glucose was noted in the fa/fa-rat islets (P = 0.13). In lean-rat islets, the glucokinase inhibitor mannoheptulose (3 mM) decreased Vmax. by 80% and increased the apparent Km from 3.3 +/- 0.7 mM to 12.2 +/- 2.0 mM (P < 0.05). There was no difference in Km or Vmax. in mannoheptulose-treated versus control islets from fa/fa rats. This lack of effect was consistent with reported effects of mannoheptulose on insulin secretion from fa/fa-rat islets [Chan, MacPhail and Mitton (1993) Can. J. Physiol. Pharmacol. 71, 34-39]. The data from glucose and mannoheptulose experiments support the hypothesis that glucokinase function is altered in fa/fa Zucker rats and may contribute to fasting hyperinsulinaemia in vivo in these animals. PMID- 8240277 TI - Characterization of a complement-fragment-C5a-stimulated calcium-influx mechanism in U937 monocytic cells. AB - The mechanism by which complement fragment C5a elevates intracellular Ca2+ ([Ca2+]i) levels in two cell types, a monocytic cell line, U937, and neutrophils, has been investigated by the use of fluorometric and radiometric techniques. In U937 cells the influx of extracellular Ca2+ can be distinguished from the release of intracellular Ca2+ stores in terms of dose-responsiveness to C5a and sensitivity to pertussis-toxin poisoning. This suggests that the mechanism of Ca2+ influx in these cells is at least partially independent of both the production of inositol phosphates and elevation of internal Ca2+ concentration. The C5a-stimulated influx of 45Ca2+ into U937 cells is inhibited by a series of metal ions (Zn2+ > Co2+ > Mn2+ > Sr2+ approximately equal to Ni2+ > La3+). The stimulated influx of Ca2+ into neutrophils is inhibited differently (Ni2 >> Co2+ > Zn2+ approximately equal to La3+ > Mn2+ approximately equal to Sr2+), is less sensitive to C5a and both the influx of extracellular Ca2+ and the release of intracellular stores are equally sensitive to pertussis toxin treatment. Taken together these results indicate that [Ca2+]i is controlled in U937 monocytes by mechanisms distinct from those which appear to operate in other myeloid cells, such as neutrophils, stimulated with C5a and formylpeptide. PMID- 8240278 TI - Spontaneously hypertensive rats and platelet Ca(2+)-ATPases: specific up regulation of the 97 kDa isoform. AB - The use of platelets instead of smooth muscle cells (SMC) to study the abnormal Ca2+ handling found in hypertension was investigated using spontaneously hypertensive rats (SHR). We studied the regulation of platelet Ca(2+)-ATPases, as we have recently demonstrated that human platelets, like SMC, contain the Ca(2+) ATPase isoform termed SERCA2-b (sarco-endoplasmic reticulum Ca(2+)-ATPase). In mixed membranes isolated from platelets of normotensive Wistar-Kyoto (WKY) rats and SHR, total Ca(2+)-ATPase activity was found to be 43% higher in SHR than in WKY rats. By the use of autophosphorylation of rat platelet Ca(2+)-ATPases with [gamma-32P]ATP, followed by SDS/PAGE and Western blotting, we found that rat platelets express two distinct Ca(2+)-ATPases: a 100 kDa isoform, recognized by a SERCA2-b-specific anti-peptide antibody, and a 97 kDa isoform, specifically recognized by a polyclonal anti-SERCA antibody. Comparative analysis of platelet membrane Ca(2+)-ATPases from WKY rats and SHR demonstrated that the expression of the SERCA2-b isoform did not change significantly (128 +/- 22%), whereas that of the 97 kDa isoform reached 300 +/- 35% in SHR when compared with WKY rats. We concluded that the upregulation of total platelet Ca(2+)-ATPases in SHR is not due to the 100 kDa SERCA2-b isoform found in SMC, but is specific to the 97 kDa Ca(2+)-ATPase isoform which is not present in SMC. Therefore platelets should be used with extreme caution as a surrogate model of vascular smooth muscle Ca2+ homeostasis. PMID- 8240279 TI - A fibroblast protein binds the 3'-untranslated region of pro-alpha 1(I) collagen mRNA. AB - Post-transcriptional regulation of the expression of the pro alpha 1(I) chain of type I collagen (COL1A1) was studied by analysing cytoplasmic RNA-binding proteins and by transient transfections with collagen minigene plasmids. In this paper we present evidence for a factor from NIH 3T3 cells and human skin fibroblasts that interacts with the conserved 3'-untranslated region (UTR) of the shorter 4.8 kb mRNA species of the COL1A1 gene. The specificity of the interaction was confirmed by using (i) unlabelled specific and non-specific competitor RNAs and (ii) oligodeoxyribonucleotides annealed to the probe or used as single-stranded competitors. An antisense oligonucleotide annealed to the RNA probe near its 3'-terminus [20-42 nucleotides upstream of the first polyadenylation signal of the alpha 1(I) collagen mRNA] inhibited the binding, whereas other sense or antisense oligonucleotides had no effect on the interaction. The binding was sensitive to alkylation of free SH groups but not to phosphatase treatment of the extracts. In u.v. cross-linking analysis this factor migrated as a single polypeptide chain of about 67 kDa, and was named alpha 1 RBF67 (type I collagen alpha 1 chain RNA-binding factor). Dexamethasone treatment of fibroblasts, which is known to accelerate the turnover of COL1A1 mRNA, decreased the alpha 1-RBF67 activity markedly as evaluated by gel-retardation and u.v. cross-linking assays. Transient transfections with plasmids carrying the alpha 1(I) collagen promoter and 3'-UTR sequences demonstrated that the 3'-UTR participates in the response to dexamethasone. Thus the loss of alpha 1-RBF67 activity might be associated with decreased alpha 1(I) collagen mRNA levels after dexamethasone treatment. PMID- 8240280 TI - Inositol phospho-oligosaccharides from rat fibroblasts and adipocytes stimulate 3 O-methylglucose transport. AB - Inositol phospho-oligosaccharides (IPOs), which are released from liver membranes upon stimulation by insulin, mimic a wide spectrum of insulin effects in different cells, but not the stimulation of glucose transport. We investigated whether other insulin-sensitive tissues release glucose transport-stimulating IPOs and whether this is related to the human insulin receptor isoform-A or -B (HIR-A or HIR-B). Rat1 fibroblasts overexpressing HIR-A or -B (rat1-HIR cells) were labelled with [3H]glucosamine, [3H]mannose or myo-[3H]inositol. IPOs from the cell supernatant were partially purified by an AG1X2 anion-exchange column, and fractions were eluted at different pH values (pH 3, pH 2 and pH 1.3). The label from glucosamine, mannose and myo-inositol appeared predominantly in the pH 2 fraction. The biological activity of the fractions was determined by measuring 3-O-methylglucose transport and lipogenesis in fat cells. Using the pH 2 fraction from the supernatant of rat1-HIR fibroblasts, insulin increased the release of 3 O-methylglucose-transport-stimulating activity (HIR-A: without insulin, 22.4 +/- 5.4%; with insulin 54.0 +/- 8.4%; HIR-B: without insulin 21.6 +/- 7.5%, with insulin, 44.7 +/- 10.6%, given as a percentage of equilibrium glucose transport reached after 4 s) and lipogenesis-stimulating activity (HIR-A: without insulin, 1.24 +/- 0.17; with insulin, 4.69 +/- 0.2; HIR-B: without insulin, 1.34 +/- 0.18; with insulin, 4.98 +/- 0.31, given as nmol of [3H]glucose converted into lipids/min per 10(6) cells). Analogous experiments were performed with isolated rat fat cells expressing the physiological level of insulin receptors. Upon insulin stimulation of fat cells in the presence of 2.5 mM mannose, the release of 3-O-methylglucose-transport-stimulating activity was detected (for purified supernatant of adipocytes without insulin, 6.9 +/- 1.12%; with insulin, 41.0 +/- 3.6%) and lipogenesis-stimulating activity (without insulin, 0.93 +/- 0.17, with insulin 2.96 +/- 0.31 nmol/min per mg). These data suggest (1) that adipocytes and rat1-HIR fibroblasts release IPOs that are able to stimulate glucose transport, (2) that both insulin receptor isoforms (HIR-A and HIR-B) mediate the effect of insulin on IPO release, and (3) that overexpression of insulin receptors increases the basal release of IPOs. PMID- 8240281 TI - Interactions between active-site-serine beta-lactamases and mechanism-based inactivators: a kinetic study and an overview. AB - The interactions between three class A beta-lactamases and three beta-lactamase inactivators (clavulanic acid, sulbactam and olivanic acid MM13902) were studied. Interestingly, the interaction between the Streptomyces cacaoi beta-lactamase and clavulanate indicated little irreversible inactivation. With sulbactam, irreversible inactivation was found to occur with the three studied enzymes, but no evidence for transiently inactivated adducts was found. Irreversible inactivation of the S. albus G and S. cacaoi enzymes was particularly slow. With olivanate, irreversible inactivation was also observed with the three enzymes, but with the S. cacaoi enzyme, no hydrolysis could be detected. A tentative summary of the results found in the literature is also presented (including 6 beta-halogenopenicillanates), and the general conclusions underline the diversity of the mechanisms and the wide variations of the rate constants observed when class A beta-lactamases interact with beta-lactamase inactivators, in agreement with the behaviours of the same enzymes towards their good and poor substrates. PMID- 8240282 TI - Met-8 of the beta 1-bungarotoxin phospholipase A2 subunit is essential for the phospholipase A2-independent neurotoxic effect. AB - beta 1-Bungarotoxin consists of a phospholipase A2 subunit and a non phospholipase A2 subunit. The toxin was oxidized with a 100-fold molar excess of chloramine T with respect to the methionine content of the protein in 0.1 M Tris/HCl at pH 8.5 and at room temperature. Reactivities of the two methionine (Met-6 and Met-8 of the phospholipase A2 subunit), five histidine, 14 tyrosine and one tryptophan residues of one toxin molecule with chloramine T were assessed from the change in intrinsic fluorescence and amino acid composition of the protein. Met-8 and one tyrosine on the phospholipase A2 subunit and less than one histidine were oxidized, while Met-6 remained intact after 30 min of reaction. One histidine and approx. two tyrosine residues were oxidized when both methionine residues were oxidized after 90 min of reaction. The sole tryptophan was oxidized slightly throughout the reaction. The chloramine T oxidation did not destroy the two Ca(2+)-binding domains, though it modified the toxin to become less effective at binding Ca2+. The modified toxin obtained after 30 or 90 min reaction time retained 65% or 40% of the phospholipase A2 activity of the parent toxin, but both were not lethal to mice and showed a very weak ability to induce the indirectly evoked contraction of chick biventer cervicis muscle. It is suggested that Met-8 may play an important role in the phospholipase A2 independent interaction with the nerve terminal membrane during the neurotoxic effect of beta 1-bungarotoxin. PMID- 8240283 TI - Purification and biochemical characterization of recombinant human placental growth hormone produced in Escherichia coli. AB - The hGH-V (or hGH-2) gene codes for human placental growth hormone (hPGH). Secretion of hPGH is continuous, in contrast with the pulsed secretion of pituitary growth hormone (hGH) which it progressively replaces in the maternal bloodstream. hGH-V cDNA has previously been cloned and isolated. Analysis of its nucleotide sequence has revealed a 191-residue protein, hPGH, differing from hGH at 13 positions. The calculated pI is more basic than that of the pituitary hormone. Here we have inserted hGH-V cDNA into the pIN-III-ompA3 plasmid in order to produce hPGH in its native form in Escherichia coli D1210. Expression of hGH-V cDNA in E. coli is significantly lower than that of hGH cDNA with the same expression system. The hPGH produced in E. coli was purified in quantities sufficient to allow its biochemical and immunochemical characterization. The molecular mass of the protein was determined by electrospray m.s. The determined mass, 22,320 Da, agrees well with the molecular mass calculated from the translated cDNA sequence, assuming the presence of two disulphide bridges. Having established the technique for producing hPGH with a primary structure identical to the natural, non-glycosylated, 22 kDa isoform, we can now plan the full physicochemical and pharmaceutical characterization of this new hormonal entity. PMID- 8240284 TI - Interaction of thrombospondin with platelet glycoproteins GPIa-IIa and GPIIb IIIa. AB - The interaction of human thrombospondin (TSP) with GPIa-IIa and GPIIb-IIIa was studied. The binding for both proteins became time-independent after 60 min. A 7 fold excess concentration of unlabelled GPIa-IIa added either initially, or after time-dependent binding, resulted in a 50% inhibition of GPIa-IIa bound to TSP. GPIa-IIa and GPIIb-IIIa specifically bound TSP since: (a) the binding of GPIIb IIIa to TSP was dependent on the presence of 1 mM MgCl2 and 1 mM CaCl2, whereas binding of GPIa-IIa was ion-independent. (b) The binding was saturable, with dissociation constants of 0.69 +/- 0.17 microM and 3.77 +/- 1.02 microM for GPIa IIa and GPIIb-IIIa respectively. (c) GPIIb-IIIa and GPIa-IIa did not significantly bind to BSA. (d) GPIIb-IIIa bound fibrinogen ion-specifically, whereas little or no binding of GPIa-IIa was detectable. (e) Both GPIIb-IIIa and GPIa-IIa bound collagen in an ion-independent manner. (f) GPIIb-IIIa did not compete with GPIa-IIa for binding to TSP. (g) Binding of GPIa-IIa to TSP was inhibited with anti-(GPIa-IIa) (6F1), whereas mouse IgG and anti-(GPIIb-IIIa) (AP 2) had no effect. (h) The interaction of GPIa-IIa with TSP is 5.5-fold more favourable than that of GPIIb-IIIa suggesting that GPIa-IIa may be a preferred binding protein for TSP-mediated platelet adhesion. PMID- 8240285 TI - Diurnal patterns of cardiac and hepatic pyruvate dehydrogenase complex activity in gold-thioglucose-obese mice. AB - The diurnal pattern of the activity of the pyruvate dehydrogenase complex (PDHC) was studied in the heart and liver of gold-thioglucose (GTG)-obese mice and age matched controls. The diurnal pattern of lipogenesis was also measured in the liver. Both lean and obese mice had one main eating period, from 20:00 to 24:00 h. Eating produced no change in serum glucose of control mice but there was a significant rise in serum insulin and triacylglycerols. There was also a 3-fold increase in cardiac PDHC activity and a 3-fold increase in hepatic lipogenesis in the control mice, but little change in hepatic PDHC activity. GTG-obese mice were hyperglycaemic, hyperinsulinaemic and hypertriglyceridaemic at all times studied, with significant increases in these parameters being seen in response to eating. Eating produced little change in cardiac PDHC activity, but there was a 5-fold increase in hepatic PDHC activity, paralleled by a 10-fold increase in hepatic lipogenesis. Hepatic PDHC activity was significantly higher in GTG-obese mice at all times except 16:00 h. The simultaneous rise of hepatic PDHC activity, lipogenesis and serum triacylglycerols in GTG-obese mice suggests an increased utilization of glucose for lipogenesis. The lack of change in heart PDHC activity in GTG-obese mice over 24 h suggests that a general decrease in PDHC activity may contribute to the development of the glucose intolerance and insulin resistance of obesity and non-insulin-dependent diabetes. However, it appears that a different level of metabolic control allows hepatic PDHC activity of the same obese animals to increase in response to hyperinsulinaemia and contribute to the higher rates of lipogenesis seen in obese mice. PMID- 8240286 TI - Dissociation of the tetrameric phosphoglycerate mutase from yeast by a mutation in the subunit contact region. AB - Phosphoglycerate mutases from different sources exhibit a variety of quaternary structures (tetramer, dimer and monomer). To perturb the tetrameric structure of yeast phosphoglycerate mutase we have prepared a mutant enzyme in which Lys-168 in the subunit-contact region has been replaced by proline. The K168P mutant enzyme undergoes dissociation to dimers at low concentrations; thus on lowering the concentration from 200 micrograms/ml to 5 micrograms/ml the proportion of tetramer falls from 85% to 53%. The tetrameric structure of the wild-type enzyme remains intact over this range of concentrations. The mutant enzyme has similar kinetic properties to the wild-type enzyme, with kcat. being reduced by 26%. Far u.v. c.d. studies show that there has been a small loss of helical structure in the mutant. Compared with wild-type enzyme, the K168P mutant enzyme is slightly less stable towards proteolysis by trypsin, but significantly less stable towards denaturation by guanidinium chloride, with the midpoint concentration of guanidinium chloride some 50% lower. After denaturation, the mutant enzyme could regain activity and quaternary structure when the guanidinium chloride concentration was lowered to 0.05 M. The properties of the mutant enzyme are discussed in terms of other dimeric phosphoglycerate and bisphosphoglycerate mutases which contain proline at position 168. PMID- 8240287 TI - Regulation of the glutamate transporter by amino acid deprivation and associated effects on the level of EAAC1 mRNA in the renal epithelial cell line NBL-I. AB - The glutamate transport system of the bovine renal epithelial cell line NBL-1 was studied. The Km for Na(+)-dependent glutamate transport was found to be 13.8 +/- 2.4 microM (Vmax. 365 +/- 19.2 pmol/3 min per mg) and for Na(+)-dependent aspartate transport 4.5 +/- 1.1 microM (Vmax. 108 +/- 6.3 pmol/3 min per mg). The Km values are in close agreement with those expected for high-affinity Na(+) dependent glutamate transport by System XAG-. Upon deprivation of amino acids, the Vmax. for Na+/aspartate co-transport rose to 203 +/- 6.0 pmol/3 min per mg (Km 3.8 +/- 0.5 microns). A probe was constructed to the high-affinity excitatory amino acid carrier (EAAC1) [Kanai and Hediger (1992) Nature (London) 360, 467 471]. The probe hybridized to a 3.5 kb transcript. On deprivation of amino acids, the level of EAAC1 mRNA decreased sharply before the measurable increase in transport levels, but was subsequently restored to control levels. A motif, which we propose is linked to amino acid deprivation, was found in the EAAC1 primary sequence. PMID- 8240288 TI - Farnesol inhibits phosphatidylcholine biosynthesis in cultured cells by decreasing cholinephosphotransferase activity. AB - The mechanism of inhibition of phosphatidylcholine (PC) biosynthesis by the isoprenoid farnesol was investigated in the human leukaemic CEM-C1 cell line. Cells were preincubated with 20 microM farnesol for up to 2 h and pulsed with [3H]choline. PC biosynthesis was inhibited to one-quarter at the step catalysed by cholinephosphotransferase (CPT). CPT activity in cellular homogenates from farnesol-treated cells was significantly decreased, but no changes in cytidylyltransferase activity or diacylglycerol concentration were observed. Measurements of CPT activity in the experiments in which farnesol was added directly to the homogenates or microsomal fractions demonstrated that farnesol did not affect CPT activity. However, cytosol from farnesol-treated samples decreased microsomal CPT activity almost twice as much as did cytosol from controls. This effect was found to be heat-stable, and disappeared after dialysis, but could not be attributed to farnesol present in the cytosol. The effect of farnesol was specific when compared with other structurally similar isoprenoids. We conclude that farnesol brings about changes in cultured cells, leading to decreased CPT activity, and thus to the inhibition of PC biosynthesis. PMID- 8240289 TI - Epidermal growth factor and phorbol myristate acetate increase expression of the mRNA for cytosolic phospholipase A2 in glomerular mesangial cells. AB - We have previously shown that phospholipase A2 (PLA2) activity is rapidly activated by epidermal growth factor (EGF) and phorbol 12-myristate 13-acetate (PMA) in renal mesangial cells and other cell systems in a manner that suggests a covalent modification of the PLA2 enzyme(s). This PLA2 activity is cytosolic (cPLA2) and is distinct from secretory forms of PLA2, which are also stimulated in mesangial cells in response to cytokines and other agonists. However, longer term regulation of cPLA2 in renal cells may also occur at the level of gene expression. Cultured rat mesangial cells were used as a model system to test the effects of EGF and PMA on the regulation of cPLA2 gene expression. EGF and PMA both produced sustained increases in cPLA2 mRNA levels, with a parallel increase in enzyme activity over time. Inhibition of protein synthesis by cycloheximide increased basal cPLA2 mRNA accumulation in serum-starved mesangial cells, and the combination of EGF and cycloheximide resulted in super-induction of cPLA2 gene expression compared with EGF alone. Actinomycin D treatment entirely abrogated the effect of EGF on cPLA2 mRNA accumulation. These findings suggest that regulation of cPLA2 is achieved by factors controlling gene transcription and possibly mRNA stability, in addition to previously characterized posttranslational modifications. PMID- 8240290 TI - Nerve growth factor activates calcium-insensitive protein kinase C-epsilon in PC 12 rat pheochromocytoma cells. AB - Protein kinase C (PKC) family members were examined in PC-12 rat pheochromocytoma cells to evaluate their role in the action of nerve growth factor (NGF). Immunoblot analysis of whole cell lysates using antibodies against various PKC isoforms revealed that PC-12 cells contained PKC-alpha, -delta, -epsilon and zeta. Assay of the protein kinase activity in these different anti-PKC immunoprecipitates demonstrated that NGF stimulated the kinase activity of PKC epsilon, but not PKC-alpha, -delta and -zeta. Both histone phosphorylation and autophosphorylation of PKC-epsilon were increased by treatment of PC-12 cells with NGF. This increased phosphorylation observed in vitro is rapid, occurring maximally at 2.5 min and declining thereafter. Moreover, this effect of NGF is dose-dependent over physiological concentrations of the growth factor. Although the mechanistic basis for this specificity in PKC activation is not clear, NGF acutely stimulated the production of diacylglycerol without causing corresponding changes in intracellular Ca2+ concentrations. These results suggest that NGF may selectively stimulate the Ca(2+)-insensitive epsilon isoform of PKC by a phosphatidylinositol-independent mechanism. PMID- 8240291 TI - Complement-induced Ca2+ influx in cultured fibroblasts is decreased by the calcium-channel antagonist nifedipine or by some bivalent inorganic cations. AB - The effects of different extracellular cations or organic Ca(2+)-channel modulators on complement-induced changes in intracellular Ca2+ and cell death have been investigated in the transfected NIH-3T3 HIR 3.5 cell line, which overexpresses the human insulin receptor. Cells were incubated with mouse anti (human insulin receptor) monoclonal antibodies before exposure to rabbit or human serum (sources of heterologous complement). Changes in intracellular Ca2+ were complement-dependent (measured by influx of 45Ca), as was cytotoxicity (monitored by leakage of lactate dehydrogenase into the culture supernatant). Addition of a dihydropyridine Ca(2+)-channel antagonist (nifedipine) or some bivalent inorganic cations caused inhibition of 45Ca entry via a novel channel distinct from endogenous voltage-gated Ca2+ channels. Nifedipine decreased, but conversely the addition of a phenylalkylamine Ca(2+)-channel antagonist (verapamil) or the inorganic Ca2+ agonists Ba2+ and Sr+ increased, complement-induced cytotoxicity. These agents had no effect on cell viability at the studied concentrations, in the absence of complement. It is concluded that complement-induced cytotoxicity is mediated by Ca2+ influx through novel specific transmembrane channels which are sensitive to the Ca(2+)-channel antagonist nifedipine, but otherwise show little resemblance to L- or T-type voltage-gated Ca2+ channels. PMID- 8240292 TI - Stimulation by paraquat of microsomal and cytochrome P-450-dependent oxidation of glycerol to formaldehyde. AB - Glycerol can be oxidized to formaldehyde by microsomes in a reaction that is dependent on cytochrome P-450. An oxidant derived from the interaction of H2O2 with iron was responsible for oxidizing the glycerol, with P-450 suggested to be necessary to produce H2O2 and reduce non-haem iron. The effect of paraquat on formaldehyde production from glycerol and whether paraquat could replace P-450 in supporting this reaction were studied. Paraquat increased NADPH-dependent microsomal oxidation of glycerol; the stimulation was inhibited by glutathione, catalase, EDTA and desferrioxamine, but not by superoxide dismutase or hydroxyl radical scavengers. The paraquat stimulation was also inhibited by inhibitors, substrate and ligand for P-4502E1 (pyrazole-induced P-450 isozyme), as well as by anti-(P-4502E1) IgG. These results suggest that P-450 still played an important role in glycerol oxidation, even in the presence of paraquat. Purified NADPH cytochrome P-450 reductase did not oxidize glycerol to formaldehyde; some oxidation, however, did occur in the presence of paraquat. Reductase plus P 4502E1 oxidized glycerol, and a large stimulation was observed in the presence of paraquat. Rates in the presence of P-450, reductase and paraquat were more than additive than the sums from the reductase plus P-450 and reductase plus paraquat rates, suggesting synergistic interactions between paraquat and P-450. These results indicate that paraquat increases oxidation of glycerol to formaldehyde by microsomes and reconstituted systems, that H2O2 and iron play a role in the overall reaction, and that paraquat can substitute, in part, for P-450 in supporting oxidation of glycerol. However, cytochrome P-450 is required for elevated rates of formaldehyde production even in the presence of paraquat. PMID- 8240293 TI - The presence and subcellular distribution of sterol carrier protein 2 in embryonic-chick tissues. AB - Transport of lipids from the yolk to the tissues of the chick embryo is slow during the first 2 weeks of development, but increases abruptly during the last week. Evidence suggests that the lipid traverses the cytoplasm of the yolk-sac membrane before secretion as lipoprotein into the fetal circulation. Little is known about the cytoplasmic transport of lipid in avian systems, but recently the presence of sterol carrier protein 2 (SCP2) was reported in chicken liver. Here we examine the cells of yolk-sac membrane, liver and small intestine for the presence of this protein as a function of the time of embryonic development. The quantity of SCP2 present in the embryonic cells did not appear to correlate with the rate of lipid flux in these tissues. The abrupt appearance of a high molecular-mass form of SCP2 was detected in small intestine shortly before hatching, but the significance of this protein is not clear. The presence of SCP2 in these tissues was also confirmed by immunocytochemical techniques. Similarly to SCP2 of mammalian cells, avian SCP2 is localized in both peroxisome-like structures and mitochondria. To a lesser extent it is associated with the endoplasmic reticulum. PMID- 8240294 TI - The substrate specificity of brain microsomal phospholipase D. AB - Neurotransmitters activate a phospholipase D that is though to specifically hydrolyse phosphatidylcholine. This enzyme has a unique property known as transphosphatidylation: in the presence of an appropriate nucleophilic receptor such as an alcohol, phospholipase D will catalyse the production of phosphatidyl alcohol. We have studied phospholipase D using an in vitro assay that uses [3H]butanol of high specific radioactivity (15 Ci/mmol) as an acceptor. In the presence of [3H]butanol and phosphatidylcholine, a microsomal membrane fraction from rat brain catalysed the production of phosphatidyl[3H]butanol. Phospholipase D activity was dependent upon the presence of a detergent; the optimal sodium oleate concentration was between 4 and 6 mM. The RF of the phosphatidyl[3H]butanol on t.l.c. was identical to the RF of the phosphatidylbutanol formed when [3H]phosphatidylcholine was incubated with 100 mM butanol. These data confirm the identity of phosphatidyl[3H]butanol. One important advantage of this assay is that the substrate does not need to be labelled. We have used this advantage to examine the substrate specificity of phospholipase D. Microsomal phospholipase D appears to hydrolyse phosphatidylcholine most efficiently. There is a relatively small but significant activity against phosphatidylethanolamine and phosphatidylserine, and there is no significant activity against phosphatidylinositol. As the head-group becomes more like choline, the phospholipid becomes a better substrate for phospholipase D. The addition of one methyl group leads to a large increase in activity. Fatty acid composition does not play a role in determining the substrate specificity. This assay should be useful in furthering our understanding of this important enzyme. PMID- 8240295 TI - F1F0-ATP synthase from bovine heart mitochondria: development of the purification of a monodisperse oligomycin-sensitive ATPase. AB - A new procedure for the isolation of ATP synthase from bovine mitochondria has been developed, with the primary objective of producing enzyme suitable for crystallization trials. Proteins were extracted from mitochondrial membranes with dodecyl-beta-D-maltoside, and the ATP synthase was purified from the extract in the presence of the same detergent by a combination of ion-exchange and gel filtration chromatography and ammonium sulphate precipitation. This simple and rapid procedure yields 20-30 mg of highly pure and monodisperse enzyme, evidently consisting of 14 different subunits, amongst them, in apparently stoichiometric amounts with the established subunits, subunit e, a recently discovered subunit of unknown function. The enzyme preparation has an oligomycin-sensitive ATP hydrolysis activity, and so the F1 domain is functionally associated with the membrane domain, F0. In contrast with the N-termini of some of the subunits of bovine mitochondrial F1-ATPase, those of the F1F0-ATP synthase are not degraded by proteolysis during the isolation procedure. This preparation therefore satisfies prerequisites for crystallization trials. PMID- 8240296 TI - Post-translational processing of progastrin: inhibition of cleavage, phosphorylation and sulphation by brefeldin A. AB - The precursor for the acid-stimulating hormone gastrin provides a useful model for studies of post-translational processing because defined sites of cleavage, amidation, sulphation and phosphorylation occur within a dodecapeptide sequence. The factors determining these post-translational processing events are still poorly understood. We have used brefeldin A, which disrupts transport from rough endoplasmic reticulum to the Golgi complex, to examine the mechanisms of cleavage, phosphorylation and sulphation of rat progastrin-derived peptides. Biosynthetic products were detected after immunoprecipitation using antibodies specific for the extreme C-terminus of progastrin, followed by reversed-phase and ion-exchange h.p.l.c. Gastrin cells incorporated [3H]tyrosine, [32P]phosphate and [35S]sulphate into both progastrin and its extreme C-terminal tryptic (nona-) peptide. Ion-exchange chromatography resolved four forms of the C-terminal tryptic fragment of progastrin which differed in whether they were phosphorylated at Ser96, sulphated at Tyr103, both or neither. The specific activity of [3H]tyrosine in the peak that was both phosphorylated and sulphated was higher than in the others. Brefeldin A inhibited the appearance of [3H]tyrosine-labelled C-terminal tryptic fragment but there was an accumulation of labelled progastrin and a peptide corresponding to the C-terminal 46 residues of progastrin. Brefeldin A also inhibited incorporation of 32P and 35S into both progastrin and its C-terminal fragment. Thus phosphorylation of Ser96, sulphation of Tyr103 and cleavage at Arg94-Arg95 depend on passage of newly synthesized progastrin along the secretory pathway; as brefeldin A is thought to act proximal to the trans Golgi, these processing steps would appear to occur distal to this point. The data also indicate that the stores of unphosphorylated C-terminal tryptic fragment are not available for phosphorylation, implying that this modification occurs proximal to the secretory granule; cleavage is known to occur in the secretory granule which suggests that it occurs after phosphorylation. PMID- 8240297 TI - Evidence for allosteric regulation of succinyl-CoA synthetase. AB - We have previously reported that distinctly different concentrations of GDP stimulate the phosphorylation and dephosphorylation of p36, the alpha-subunit of succinyl-CoA synthetase (SCS) in Dictyostelium discoideum. In this present study, we have investigated the mechanism underlying these dual effects of GDP. Dephosphorylation of p36 is induced by relatively high levels of GDP and is coincident with the formation of GTP. This indicates that, at high concentrations, GDP serves as a substrate of SCS. However, 100-fold lower concentrations of GDP, which do not bind to the catalytic site to induce SCS dephosphorylation, stimulate p36 phosphorylation. This stimulation is not diminished by dilution of the sample, and is retained during purification of the protein. Gel-filtration analyses indicate that SCS in our system behaves as a non interacting alpha beta dimer, the hydrodynamic behaviour of which is not altered by the presence of added GDP. The data indicate that altered protein-protein interactions do not account for the stimulation of p36 phosphorylation by low GDP concentrations. We propose that GDP functions as an allosteric regulator of SCS, and experiments using guanosine 5'-[beta-thio]diphosphate (GDP[S]) are shown to distinguish further the allosteric and catalytic binding sites. PMID- 8240298 TI - Structural features responsible for kinetic thermal stability of a carboxypeptidase from the archaebacterium Sulfolobus solfataricus. AB - Investigations were performed on the structural features responsible for kinetic thermal stability of a thermostable carboxypeptidase from the thermoacidophilic archaebacterium Sulfolobus solfataricus which had been purified previously and identified as a zinc metalloprotease [Colombo, D'Auria, Fusi, Zecca, Raia and Tortora (1992) Eur. J. Biochem. 206, 349-357]. Removal of Zn2+ by dialysis led to reversible activity loss, which was promptly restored by addition of 80 microM ZnCl2 to the assay mixture. For the first-order irreversible thermal inactivation the metal-depleted enzyme showed an activation energy value of 205.6 kJ.mol-1, which is considerably lower than that of the holoenzyme (494.4 kJ.mol-1). The values of activation free energies, enthalpies and entropies also dropped with metal removal. Thermal inactivation of the apoenzyme was very quick at 80 degrees C, whereas the holoenzyme was stable at the same temperature. These findings suggest a major stabilizing role for the bivalent cation. Chaotropic salts strongly destabilized the holoenzyme, showing that hydrophobic interactions are involved in maintaining the native conformation of the enzyme. However, the inactivation rate was also increased by sodium sulphate, acetate and chloride, which are not chaotropes, indicating that one or more salt bridges concur in stabilizing the active enzyme. Furthermore, at the extremes of the pH-stability curve, NaCl did not affect the inactivation rate, confirming the stabilizing role of intramolecular ionic bonds, as a pH-dependent decrease in stability is likely to occur from breaking of salt bridges involved in maintaining the native conformation of the protein. PMID- 8240299 TI - Intact transmembrane isoforms of the neural cell adhesion molecule are released from the plasma membrane. AB - Three soluble neural cell adhesion molecule (NCAM) polypeptide classes of M(r) values 190,000 (NCAM-s1), 135,000 (NCAM-s2) and 115,000-110,000 (NCAM-s3) have been demonstrated in rat brain and cerebrospinal fluid [Krog, Olsen, Dalseg, Roth and Bock (1992) J. Neurochem. 59, 838-847]. NCAM-s3 is known to arise from released glycosylphosphatidylinositol (GPI)-linked NCAM [He, Finne and Goridis (1987) J. Cell. Biol. 105, 2489-2500] as well as from extracellularly cleaved transmembrane NCAM isoforms [Nybroe, Linnemann and Bock (1989) J. Neurochem. 53, 1372-1378]. In this study the origin of NCAM-s1 and NCAM-s2 and the function of soluble NCAM forms were investigated. It was shown that all three soluble forms could be released from brain membranes with M(r) values identical to the three major membrane-associated forms: the large transmembrane 190,000-M(r) form (NCAM A), the smaller transmembrane 135,000-M(r) form (NCAM-B) and the GPI-anchored 115,000-110,000-M(r) form (NCAM-C). A polyclonal antibody, directed against transmembrane and cytoplasmic epitopes common to NCAM-A and NCAM-B, was shown to react with NCAM-s1 and NCAM-s2. Furthermore, NCAM-B was shown to be shed in a presumably intact soluble form from membranes of cells transfected with this isoform. Thus, NCAM-s1 and NCAM-s2 probably represent intact released transmembrane NCAM-A and NCAM-B. The soluble transmembrane forms are likely to exist in vivo, as NCAM-s1 and NCAM-s2 were readily demonstrated in cerebrospinal fluid. By density-gradient centrifugation it was shown that shed transmembrane NCAM-B was present in fractions of high, as well as low, density, indicating that a fraction of the shed NCAM is associated with minor plasma membrane fragments. Finally, it was shown that isolated soluble NCAM inhibited cell binding to an immobilized NCAM substratum, attributing a pivotal role to soluble NCAM in vivo as a modulator of NCAM-mediated cell behaviour. PMID- 8240300 TI - Site-directed removal of N-glycosylation sites in the bovine cation-dependent mannose 6-phosphate receptor: effects on ligand binding, intracellular targetting and association with binding immunoglobulin protein. AB - The bovine cation-dependent mannose 6-phosphate receptor (CD-MPR) contains five potential N-linked glycosylation sites, four of which are utilized. To evaluate the function of these oligosaccharides, site-directed mutagenesis was used to generate glycosylation-deficient CD-MPR mutants lacking various potential glycosylation sites. The mutants were constructed in both a full-length and a soluble truncated (STOP155 construct) form of the receptor and their properties were examined in transfected COS-1 cells. The results showed that the presence of a single oligosaccharide chain, particularly at position 87, on the CD-MPR significantly enhanced its mannose 6-phosphate (Man-6-P)-binding ability when compared with non-glycosylated receptors. In addition, the presence of a single oligosaccharide chain at position 87, and to a lesser degree at position 31 or 81, promoted the secretion of the STOP155 CD-MPR. Pulse-labelling of transfected COS-1 cells followed by immunoprecipitation with binding immunoglobulin protein (BiP)-specific and CD-MPR-specific antibodies indicated that BiP associated with the non-glycosylated forms of the receptor but not with the wild-type CD-MPR. Furthermore, the association of the various glycosylation-deficient forms of the CD-MPR with BiP correlated inversely with their ability to bind Man-6-P. From these results we conclude that N-glycosylation of the bovine CD-MPR facilities the folding of the nascent polypeptide chain into a conformation that is conductive for intracellular transport and ligand binding. PMID- 8240301 TI - A 60 kDa polypeptide of skeletal-muscle sarcoplasmic reticulum is a calmodulin dependent protein kinase that associates with and phosphorylates several membrane proteins. AB - Activation of a calmodulin (CaM)-dependent protein kinase associated with rabbit skeletal-muscle sarcoplasmic reticulum (SR) results in the phosphorylation of polypeptides of 450, 360, 165, 105, 89, 60, 34 and 20 kDa. Radioligand-binding studies indicated that a membrane-bound 60 kDa polypeptide contained both CaM- and ATP-binding domains. Under renaturing conditions on nitrocellulose blots, the 60 kDa polypeptide of the membrane exhibited CaM-dependent autophosphorylation activity, suggesting that it was the CaM-dependent protein kinase of SR. Ca2+/CaM independent autophosphorylation of polypeptides of 62 and 45 kDa was found to occur in the light SR, whereas the Ca2+/CaM-dependent autophosphorylation activity was enriched in the heavy SR. Both these kinase activities were absent from transverse tubules, although these membranes were enriched in CaM-binding polypeptides of 160, 100 and 80 kDa. In the absence of Ca2+, CaM bound to a 33 kDa polypeptide of the membrane. The purified ryanodine receptor was not phosphorylated by the purified CaM kinase, although it was a substrate for protein kinase C. Affinity-purified antibodies to brain CaM kinase II cross reacted with the 60 kDa polypeptide in Western blots and immunoprecipitated the 60 kDa polypeptide, along with the 360, 105, 89, 34 and 20 kDa phosphoproteins, from Nonidet-P-40-solubilized SR membranes. Antibodies raised against the 60 kDa kinase polypeptide did not cross-react with the other phosphoproteins, suggesting that these polypeptides were distinct and unrelated. Subcellular distribution of the 60 kDa kinase indicated the specific association of the polypeptide with the junctional-face membrane of SR. The CaM-dependent incorporation of 32P into various membrane proteins was inhibited by the CaM kinase II fragment (290-309), with an IC50 value of 2 nM for the inhibition of incorporation into the 60 kDa kinase polypeptide. Recent studies [Wang and Best (1992) Nature (London) 359, 739 741] have shown that a CaM kinase activity intrinsic to the membrane can inactivate the Ca(2+)-release channel of skeletal muscle SR. Since our results demonstrate that the 60 kDa polypeptide of SR is a CaM-dependent protein kinase, we suggest that this kinase, through its associations, may be responsible for gating the Ca(2+)-release channel. PMID- 8240302 TI - Structure and biological activity of glucagon and glucagon-like peptide from a primitive bony fish, the bowfin (Amia calva). AB - The bowfin, Amia calva (order Amiiformes) occupies an important position in phylogeny as a surviving representative of a group of primitive ray-finned fishes from which the present-day teleosts may have evolved. Glucagon and glucagon-like peptide (GLP) were isolated from an extract of bowfin pancreas and their primary structures determined. Bowfin glucagon shows only four amino acid substitutions compared with human glucagon, and bowfin glucagon was equipotent and equally effective as human glucagon in stimulation of glycogenolysis in dispersed hepatocytes from a teleost fish, the copper rockfish, Sebastes caurinus. In contrast, bowfin GLP shows 15 amino acid substitutions and three amino acid deletions compared with the corresponding region of human GLP-1-(7-37)-peptide. In particular, the bowfin peptide contains an N-terminal tyrosine residue rather than the N-terminal histidine residue found in all other glucagon-related peptides so far characterized. Bowfin GLP stimulated glycogenolysis in rockfish hepatocytes, but was 3-fold less effective and 23-fold less potent than human GLP 1-(7-37)-peptide. We speculate that selective mutations in the GLP domain of bowfin preproglucagon may be an adaptive response to the previously demonstrated low biological potency of bowfin insulin. PMID- 8240303 TI - myo-inositol transport and metabolism in fetal-bovine aortic endothelial cells. AB - The myo-inositol transport system in confluent fetal-bovine aortic endothelial cells was characterized after 7-10 days in subculture, at which time the myo inositol levels and rates of myo-[2-3H]-inositol uptake and incorporation into phospholipid had reached steady state. Kinetic analysis indicated that the uptake occurred by both a high-affinity transport system with an apparent Kt of 31 microM and Vmax. of 45 pmol/min per mg of protein, and a non-saturable low affinity system. Uptake was competitively inhibited by phlorhizin, with a Ki of 50 microM; phloretin was a non-competitive inhibitor, with half-maximal inhibition between 0.2 and 0.5 mM. Glucose was a weak competitive inhibitor, with a Ki of 37 mM; galactose failed to inhibit uptake. A weak dependence on Na+ for the initial rate of uptake was observed at 11 microM myo-inositol. When fetal bovine-serum (FBS)-supplemented medium, which contained 225 microM myo-inositol, was used, the cells contained about 200 nmol of myo-inositol/mg of DNA. With adult-bovine-serum (ABS)-supplemented medium, which contained 13 microM myo inositol, the cells contained about 110 nmol/mg of DNA. Transport of 11 microM myo-[2-3]inositol was 18 and 125 pmol/min per mg of DNA for cells grown in FBS and ABS respectively. Kinetic analysis showed that for the cells grown in FBS the Vmax. of the high-affinity system was decreased by 64%, whereas the Kt remained essentially unchanged. Increased cell myo-inositol levels were not associated with an increased rate of phosphatidylinositol synthesis. After prolonged exposure of fetal endothelial cells to a myo-inositol concentration which approximated to a high fetal as opposed to a low adult blood level, cell myo inositol levels doubled and high-affinity transport underwent down-regulation. PMID- 8240304 TI - Substrate-induced inactivation of the OXA2 beta-lactamase. AB - The hydrolysis time courses of 22 beta-lactam antibiotics by the class D OXA2 beta-lactamase were studied. Among these, only three appeared to correspond to the integrated Henri-Michaelis equation. 'Burst' kinetics, implying branched pathways, were observed with most penicillins, cephalosporins and with flomoxef and imipenem. Kinetic parameters characteristic of the different phases of the hydrolysis were determined for some substrates. Mechanisms generally accepted to explain such reversible partial inactivations involving branches at either the free enzyme or the acyl-enzyme were inadequate to explain the enzyme behaviour. The hydrolysis of imipenem was characterized by the occurrence of two 'bursts', and that of nitrocefin by a partial substrate-induced inactivation complicated by a competitive inhibition by the hydrolysis product. PMID- 8240305 TI - Application of the principle of microscopic reversibility to the steady-state rate equation for a general mechanism for an enzyme reaction with substrate and modifier. PMID- 8240306 TI - Some classical errors in the kinetic analysis of enzyme reactions. PMID- 8240307 TI - Differences in 2-oxoglutarate dehydrogenase regulation in liver and kidney. PMID- 8240308 TI - Bacterial oxidative-stress substance is 2-C-methyl-D-erythritol 2,4 cyclopyrophosphate. PMID- 8240309 TI - Biotin binders selected from a random peptide library expressed on phage. PMID- 8240310 TI - Protein lateral mobility as a reflection of membrane microstructure. AB - The lateral mobility of membrane lipids and proteins is presumed to play an important functional role in biomembranes. Photobleaching studies have shown that many proteins in the plasma membrane have diffusion coefficients at least an order of magnitude lower than those obtained when the same proteins are reconstituted in artificial bilayer membranes. Depending on the protein, it has been shown that either the cytoplasmic domain or the ectodomain is the key determinant of its lateral mobility. Single particle tracking microscopy, which allows the motions of single or small groups of membrane molecules to be followed, promises not only to reveal new features of membrane dynamics, but also to help explain longstanding puzzles presented by the photobleaching studies, particularly the so-called immobile fraction. The combination of the two complementary technologies should measurably enhance our understanding of membrane microstructure. PMID- 8240311 TI - The fibrillin-Marfan syndrome connection. AB - A few years ago no one would have suspected that the well-known disorder of connective tissue, Marfan syndrome, could be caused by mutations in a recently discovered extracellular component, fibrillin. Likewise, nobody would have predicted that fibrillin represents a small family of proteins that are associated with several phenotypically overlapping disorders. The fibrillins are integral constituents of the non-collagenous microfibrils, with an average diameter of 10 nm. These aggregates are distributed in the extracellular matrix of virtually every tissue. Microfibrillar bundles provide the external coating to elastin in elastic fibers, and serve an anchoring function in non-elastic tissues. At higher resolution, individual microfibrils have a "beads-on-a-string" appearance resulting from the head-to-tail polymerization of multiple fibrillin aggregates. Structurally, fibrillin contains a series of repeated sequences homologous to the epidermal growth factor calcium-binding motif. Characterization of fibrillin mutations in Marfan syndrome patients, together with the elucidation of the structure of the fibrillin proteins, have provided new insights, and raised new questions, about the function of the 10 nm microfibrils. For example, it is possible that the fibrillins, in addition to serving a structural function, might also be involved in regulating cellular activities and morphogenetic programs. It is fitting that the long search for the Marfan syndrome gene has brought a novel group of proteins to the forefront of extracellular matrix biology. PMID- 8240312 TI - The spliceosome. AB - The spliceosome is a large RNA-protein complex that catalyses the removal of introns from nuclear pre-mRNA. A wide range of biochemical and genetical studies shows that the spliceosome comprises three major RNA-protein subunits, the U1, U2 and [U4/U6.U5] small nuclear ribonucleoprotein particles (snRNPs), and an additional group of non-snRNP protein splicing factors. Rapid progress is being made in unravelling the interactions which take place between these factors during the splicing reaction. The emerging picture of the spliceosome reveals a highly dynamic structure that assembles on pre-mRNA transcripts in a stepwise pathway and is organised, at least in part, by complex RNA base-pairing interactions between the small nuclear RNAs (snRNAs) and the intron substrate. Many of these interactions can be detected both in mammalian and yeast spliceosomes, suggesting that the basic splicing mechanism is an ancient one that has been highly conserved during evolution. PMID- 8240313 TI - Assembly of intermediate filaments. AB - The assembly of intermediate filaments is a fundamental property of the central rod domain of the individual subunit proteins. This rod domain, with its high propensity for alpha-helix formation, is the common and identifying feature of this family of proteins. Assembly occurs in vitro in the absence of other proteins or exogenous sources of energy; in vivo, it appears as if other factors, as yet poorly understood, modulate the assembly of intermediate filaments. Parallel, in-register dimers form via coiled-coil interactions of the rod domain. Tetramers may form from staggered arrays of parallel or antiparallel arrangements of dimers. Higher-order polymerization, which occurs spontaneously if the ionic strength of a mixture of dimers and tetramers is raised, proceeds rapidly through poorly described intermediates to the final 10 nm filament. This process is dependent on and modulated by the non-alpha-helical end domains, as well as those amino acids present at the very beginning and end of the rod domain. The interactions governing tetramer formation are most probably the same ones that are responsible for the lateral and longitudinal associations within intermediate filaments. PMID- 8240314 TI - Recent experiments with MHC knock-out mice: more questions than answers. PMID- 8240315 TI - Relating biochemistry to biology: how the recombinational repair function of RecA protein is manifested in its molecular properties. AB - The multiple activities of the RecA protein in DNA metabolism have inspired over a decade of research in dozens of laboratories around the world. This effort has nevertheless failed to yield an understanding of the mechanism of several RecA protein-mediated processes, the DNA strand exchange reactions prominent among them. The major factors impeding progress are the invalid constraints placed upon the problem by attempting to understand RecA protein-mediated DNA strand exchange within the context of an inappropriate biological paradigm-namely, homologous genetic recombination as a mechanism for generating genetic diversity. In this essay I summarize genetic and biochemical data demonstrating that RecA protein evolved as the central component of a recombinational DNA repair system, with the generation of genetic diversity being a sometimes useful byproduct, and review the major in vitro activities of RecA protein from a repair perspective. While models proposed for both recombination and recombinational repair often make use of DNA strand cleavage and transfer steps that appear to be quite similar, the molecular and thermodynamic requirements of the two processes are very different. The recombinational repair function provides a much more logical and informative framework for thinking about the biochemical properties of RecA and the strand exchange reactions it facilitates. PMID- 8240316 TI - Allosteric proteins: from regulatory enzymes to receptors--personal recollections. PMID- 8240317 TI - A new member of the C1s family of complement proteins found in a bactericidal factor, Ra-reactive factor, in human serum. AB - The Ra-reactive factor (RaRF) found in vertebrate sera activates the C4 and C2 components of complement. The C4/C2-activating component of mouse RaRF has been found to contain a 100-kDa serine protease called P100. In the present study, we cloned a cDNA with cDNA of mouse RaRF P100 as a probe from a human liver cDNA library. An open reading frame of 2097 nucleotides encoding a protein of 699 residues was found in the cloned cDNA of 4489 nucleotides. This protein exhibits 87.4% amino acid homology with mouse P100, and 36.4% and 37.1% homologies with that of the C1r and C1s subcomponents of human complement, respectively. The characteristic nodules and domain of C1r and C1s were highly conserved in this protein. This indicates that the P100, together with the C1r and C1s, forms a unique protein family having the same module/domain constitution. PMID- 8240318 TI - Amplification and overexpression of cyclin D1 in human hepatocellular carcinoma. AB - Amplification of the chromosome 11q13 region occurs in several types of human cancer including esophageal, breast, lung, bladder and hepatocellular carcinoma (HCC). The gene cyclin D1 maps to this region in close proximity to two proto oncogenes hst-1 and int-2. We previously demonstrated that cyclin D1 was not only amplified but also overexpressed in about 30% of human esophageal cancers. To investigate the role of cyclin D1 in human hepatocellular carcinoma (HCC), DNA from 30 HCC and 5 control liver tissues from Taiwan and also the HCC cells lines HepG2 and Hep3B, were examined for amplification of the cyclin D1 gene. A 3 to 20 fold amplification was found in 4 of the 30 (13%) HCC samples but not in any of the 5 control tissues or the 2 cell lines. Immunohistochemical analysis of cyclin D1 indicated overexpression of this protein in tumors that displayed gene amplification. Weak or negative staining was observed in the other HCC samples as well as in the control tissues and cell lines. These data suggest that increased expression of cyclin D1 may play an important role in the development of a subset of human HCC, perhaps by perturbing normal control of the cell cycle. PMID- 8240319 TI - Molecular cloning of hippocalcin, a novel calcium-binding protein of the recoverin family exclusively expressed in hippocampus. PMID- 8240320 TI - N-glycosylation of forms of angiotensin converting enzyme from four mammalian species. AB - To help clarify bases for the molecular weight and surface charge heterogeneities of forms of somatic angiotensin converting enzyme (ACE), we examined for differences in N-glycosylation. ACE preparations purified from human, guinea pig, rat and rabbit tissues were found to be heterogeneous in terms of numbers of N glycosylated sites (7-8 sites per molecule of ACE) and in types of structures of oligosaccharides used for glycosylation (complex versus high mannose oligosaccharide contents). Our findings, taken with reports of potential N glycosylation sites and amino acid sequencing data, indicate that ACE forms can differ in terms of degrees of glycosylation, sites of glycosylation and structures of attached oligosaccharide units. PMID- 8240321 TI - A comparison of guinea pig serum angiotensin converting enzyme with forms of angiotensin converting enzyme from human, rat and rabbit tissues. AB - Guinea pig serum angiotensin converting enzyme (ACE) activities exceed ACE activities of other mammalian sera by as much as two magnitudes. To examine the possibility that guinea pig ACE has a superior catalytic efficiency, we purified it to apparent homogeneity and compared it to highly purified forms of ACE from human seminal plasma, rat lungs and rabbit lungs. The first 24 amino acid residues of guinea pig and rat ACE forms were 96% identical with the sequence of human ACE. Second order rate constants (kcat/Km) for guinea pig, human and rabbit forms of ACE on reaction with benzoyl-Phe-Ala-Pro were identical (1.6E-09 M-1 min 1). Their dissociation constants on reaction with the ACE inhibitor RAC-X-65 were within a narrow range (10-16 pM). Thus, the high ACE activity of guinea pig serum is owing to high enzyme concentration and not to superior catalytic efficiency. PMID- 8240322 TI - Genotypic heterogeneity among Lewis negative individuals. AB - We have identified a C to T mutation at nucleotide No 314 in the coding region of the human alpha (1,3/1,4) fucosyltransferase III (FT-III) gene held responsible for synthesis of Lewis a and b antigens. The mutation gives rise to an additional cleavage site for restriction enzyme NlaIII and was found in both alleles in 5 of 18 Lewis negative but in none of 22 Lewis positive individuals. Heterozygous individuals were found both among Le negative (10 of 18) and Le positive (4 of 22) individuals. The mutation corresponds to a threonine to methionine substitution and was found by partial sequencing of the coding sequence of one of five Lewis negative individuals homozygous for this transition. PMID- 8240323 TI - Evidence of an intermediate-size beta-endorphin-containing precursor molecule in bovine pituitary by mass spectrometry. AB - A new analytical system has been designed to characterize intermediate-sized precursor molecules from two different precursors-the preproenkephalin Abovine,1 263 precursor molecule, which produces methionine enkephalin and from the proopiomelanocortin, POMCbovine,1-265 precursor molecule, which produces beta endorphin. A possible intermediate-sized precursor that leads to beta-endorphin was found in bovine pituitary by means of electrospray ionization mass spectrometry, in combination with gel permeation chromatography, reversed-phase high performance liquid chromatography, radioimmunoassay, trypsinolysis, and liquid secondary ion mass spectrometry. PMID- 8240324 TI - An unspliced cDNA for human dihydrolipoamide succinyltransferase: characterization and mapping of the gene to chromosome 14q24.2-q24.3. AB - Abnormality of the dihydrolipoamide succinyltransferase gene may be a cause of familial Alzheimer's disease linked to chromosome 14q24.3. However, the locus of the dihydrolipoamide succinyltransferase gene on this chromosome was uncertain. An unspliced cDNA of about 2.3 kb for human dihydrolipoamide succinyltransferase was isolated. This cDNA contained three exons and four introns and the nucleotide sequences at the 5' donor and 3' acceptor sites of all introns conformed to the gt-ag rule. The amino acid sequences of the three exons support our previous observation that human dihydrolipoamide succinyltransferase lacks a sequence motif for an E1 and/or E3 binding site. The unspliced cDNA was mapped only on human chromosome 14q24.2-q24.3 by fluorescent in situ hybridization. Thus the dihydrolipoamide succinyltransferase gene is concluded to be located on human chromosome 14q24.2-q24.3. PMID- 8240325 TI - Carboxyl methylation of the low molecular weight GTP-binding protein G25K: regulation of carboxyl methylation by rhoGDI. AB - A soluble form of G25K from brain co-purified with a 28 kDa protein, which was identified as guanine nucleotide dissociation inhibitor (GDI) protein, similar or identical to rhoGDI. The GDI protein inhibited the dissociation of GDP from G25K. G25K and the GDI protein form a heterodimer and remain associated with either GDP or GTP gamma S bound. The GDI protein inhibited carboxyl methylation of G25K in the presence of magnesium and GDP. The GDI protein appears to be an important regulator of G25K methylation by blocking methylation of G25K in the inactive GDP bound conformation. PMID- 8240326 TI - Affinity-labeling of an NADPH-binding site on the heavy subunit of flavocytochrome b558 in particulate NADPH oxidase from activated human neutrophils. AB - Cell stimulation of blood phagocytes activates the superoxide-producing NADPH oxidase. Cytochrome b558, one of the two oxidase redox components, comprises a light (alpha) and a heavy glycosylated (beta) subunit. The other redox component, a flavoprotein, is now thought to be the heavy subunit, on the basis of amino acid sequence comparisons and of reconstitution experiments with purified components. We published that pyridoxal-5'-diphospho-5'-adenosine is an inactivating affinity label for the NADPH-binding site of particulate oxidase from activated neutrophils. We have now radiolabeled the inactivated oxidase by reducing with Na[3H]BH4 the Schiff base formed between proteins and the reagent. Upon SDS-PAGE, the NADPH-inhibitable incorporation is found at the same position as the immunodetectable cytochrome heavy subunit, before and after deglycosylation. Membranes from either activated cells of a cytochrome-deficient X-linked granulomatous disease patient or normal resting cells show no incorporation at this position. Our results provide experimental evidence for the existence on the cytochrome b558 heavy chain of an NADPH-binding site which can only be affinity-labeled by PLP-AMP when the oxidase is active. This suggests the occurrence of a conformational change in the cofactor binding site upon enzyme activation. PMID- 8240327 TI - Guanylin mRNA is expressed in villous enterocytes of the rat small intestine and superficial epithelia of the rat colon. AB - Guanylin, and endogenous ligand for the Escherichia coli heat-stable enterotoxin receptor, is a recently characterized intestinal peptide. To understand the possible physiologic function of guanylin, we examined the cellular location of guanylin mRNA expression in the rat intestine. Intestinal cells were sequentially isolated from villous tip to crypt in rat jejunum and ileum. Northern blots of total RNA identified a single 0.65 kb guanylin transcript predominantly in the villous cell fractions. In situ hybridization studies demonstrated maximal signal intensity in villous cells in rat ileum and surface epithelial cells in the colon. In the ileum, the signal was nonuniform in distribution in the surface epithelial cells, with focal areas of intense signal in clusters of columnar absorptive cells. In both colon and ileum, signal intensity was near background level in deep crypt cells, lamina propria, and muscularis. PMID- 8240328 TI - Transmembrane Ca2+ gradient-mediated modulation of sarcoplasmic reticulum Ca(2+) ATPase. AB - Ca(2+)-ATPase from skeletal muscle sarcoplasmic reticulum was reconstituted into liposomes with (100-1000 fold) or without transmembrane Ca2+ gradient. The highest enzyme activity and Ca2+ uptake were observed in the vesicles without transmembrane Ca2+ gradient. If there existed transmembrane Ca2+ gradient, no matter what the direction was, a lower activity would appear. Dissipation of transmembrane Ca2+ gradient by A23187 could lead to a change in enzyme activity of incorporated Ca(2+)-ATPase. A concomitant change of lipid fluidity of proteoliposomes with that of enzyme activity and Ca2+ uptake was observed. The inhibition of Ca(2+)-ATPase by the transmembrane Ca2+ gradient could be observed in the PC-PE vesicles, but not in the PS-PE or PG-PE proteoliposomes. PMID- 8240329 TI - Rabbit skeletal muscle actin behaves differently than Acanthamoeba actin when added to soluble extracts of Acanthamoeba castellanii. AB - Cold extracts of Acanthamoeba castellanii in polymerizing buffer contain 32 microM unpolymerized actin of which about 20% polymerizes (as measured by ultracentrifugation) when the extract is warmed to 22 degrees C. As quantified by the increase in fluorescence of pyrene-labeled actin, 16% of muscle G-actin and 46% of Acanthamoeba G-actin polymerized when 0.8 microM of each was added to warm extracts of Acanthamoeba. Added muscle F-actin (1.2 microM) rapidly and totally depolymerized and then partially repolymerized whereas 1.2 microM added Acanthamoeba F-actin was stable indefinitely. Furthermore, muscle actin subunits were completely removed from copolymers of muscle and Acanthamoeba F-actin while all the amoeba actin remained polymerized when the copolymers contained at least 50% amoeba actin. These results suggest that exogenous tracer actin may not be an accurate indicator of the dynamics of endogenous actin in extracts and cells. PMID- 8240330 TI - Identification of a novel collagen chain represented by extensive interruptions in the triple-helical region. AB - We have isolated mouse cDNA clones that encode a newly discovered collagenous polypeptide. Four overlapping cDNA clones contained an open reading frame of 1140 amino acid residues of collagenous and non-collagenous domains. The predicted polypeptide consists of 10 collagenous domains of various length that are interrupted by 11 noncollagenous domains. The overall structural arrangement differed significantly from reported collagen chains of 17 different types. Northern-blot analyses showed hybridization of the cDNA to 5.6kb and 4.8kb mRNA species in liver and kidney that are due to utilization of the two poly (A) signals. PMID- 8240331 TI - Mercuric ion binding abilities of MerP variants containing only one cysteine. AB - Using site-directed mutagenesis, Cys14 and Cys17 in MerP were replaced in turn by serine or alanine. All four variants were purified and partially characterized. The The mutant proteins all had one reactive thiol group left. In the absence of external thiols, the protein variants bound between two and four Hg2+, but unlike non-mutant MerP, none of the variants could bind Hg2+ when external thiol was added. This loss of the ability to specifically bind one Hg2+ per protein molecule shows that both cysteine residues 14 and 17 are necessary for binding of Hg2+ when there is competition from other thiol groups. PMID- 8240332 TI - Stathmin in mung bean leaves and rat brain. AB - Stathmin is a highly-conserved, cytosolic protein whose synthesis and phosphorylation is closely associated with growth and differentiation. Although conserved among vertebrates, stathmin has not been identified in plants. In the present study, anti-stathmin antibodies were generated against a synthetic peptide corresponding to amino-acid residues 32-44 of rat stathmin, and these antibodies were used to probe immunoblots of proteins from rat brain and mung bean. The antibodies recognized 12-kDa, 21-kDa and 22-kDa proteins in cytosolic fractions from mung bean leaves and a 12-kDa protein in cytosolic fractions from roots. The two larger proteins identified by the antibodies have apparent molecular weights and isoelectric points similar to those of rat brain stathmin. These results are the first to show that stathmin-like proteins are present in plants. PMID- 8240333 TI - Protein oxidation in magnesium deficient rat brains and kidneys. AB - Magnesium deficiency produces a pro-oxidant systemic inflammation in rats. The purpose of these experiments was to determine if and when there is any oxidation of cellular proteins. We have found there is a significant increase in protein oxidation products, protein carbonyls, in both the brain and the kidney within 2 to 3 weeks on a magnesium deficient diet. These changes occur prior to any detectable changes in cellular glutathione, tissue damage or dysfunction. We conclude that oxidation of cellular proteins occur early in magnesium deficiency and may contribute to the tissue damage and loss of function observed in the later stages of the deficiency. This is the first demonstration of the time course of protein oxidation product development in magnesium deficient animals. PMID- 8240334 TI - Recognition of non-B DNA structures by cellular proteins. AB - By utilising a gel retardation assay, cytoplasmic extracts of Hela cells have been shown to contain a number of polypeptides which bind preferentially to non-B forms of DNA. These polypeptides vary in apparent molecular weights and are present as minor components of the total extract. Similar properties were demonstrated by purified preparations of eukaryotic topoisomerase I and by a DNA binding subunit of the transcription factor NFkB. These results suggest that non B DNA binding proteins may represent families of proteins which recognise more open forms of DNA as a cognate parameter to their cellular functions. PMID- 8240335 TI - The serum unresponsive Rous sarcoma virus promoter sustains a high serum response factor-dependent transcription in vitro. AB - CArG boxes are cis-regulatory elements which are represented both in serum responsive and unresponsive promoters. Here we show that the RSV Long Terminal Repeat contains two CArG boxes, which were efficiently recognised by purified Serum Response Factor, although they remained unresponsive to serum in transient transfection assays. However, RSV CArG boxes were as efficient as c-fos Serum Response Element in mediating a Serum Response Factor-dependent transcription in vitro. Thus, the fact that a CArG box is able to bind Serum Response Factor in an active form is insufficient for serum responsiveness in vivo. PMID- 8240336 TI - Isolation of a cDNA clone, encoding the ribosomal protein S20, downregulated during the onset of apoptosis in a human leukaemic cell line. AB - In this report we describe the isolation, by differential screening, and characterization of a cDNA clone downregulated fivefold in association with the induction of apoptosis in the human leukaemic cell line CEM C7. DNA sequence analysis identified this clone as representing the gene encoding the S20 ribosomal protein. Interestingly, the expression of the S20 mRNA was downregulated early during the induction of apoptosis in this model system, prior to the onset of DNA fragmentation and other morphological changes associated with cell death, suggesting some degree of involvement of this particular gene in the biochemical events that occur during the onset of cell death. PMID- 8240337 TI - Alpha (1,3/1,4)fucosyltransferase (FucT-III) gene is inactivated by a single amino acid substitution in Lewis histo-blood type negative individuals. AB - Human alpha (1,3/1,4)fucosyltransferase (FucT-III) genes were cloned from genomic DNAs of seven Lewis negative (Le(a- b-)) individuals. Restriction mapping analyses revealed that an additional Pvu II site was created in the FucT-III genes from Lewis negative individuals. Four of these individuals were shown to be heterozygous for the Pvu II site and three were homozygous for it. The Pvu II site was produced by a single base substitution at the 508 nucleotide of coding sequence, G to A, resulting in the amino acid substitution, Gly to Ser at position 170. Another single base substitution, resulting in the change of Asp to Ala at position 336, was observed in the Lewis negative FucT-III gene which did not have the Pvu II site. The FucT-III enzyme was inactivated by these two mutations resulting in a single amino acid substitution in the catalytic region. All FucT-III genes, regardless of the Pvu II site, from Lewis negative individuals had a single base substitution, the T at position 59 to G, resulting in the change of Leu to Arg at position 20. This mutation, however, was not responsible for FucT-III gene inactivation. PMID- 8240338 TI - Metallothionein reacts with Fe2+ and NO to form products with A g = 2.039 ESR signal. AB - The interaction of metallothionein, Fe2+, and nitric oxide has been examined by electron spin resonance spectroscopy. When either apoMT 2 or Zn-MT 2 from rabbit liver was mixed with Fe2+ and NO, paramagnetic products with g values of 2.013 and 2.039 were found. These are characteristic of iron nitrosyl thiolates with stoichiometry, Fe(NO)2(SR)2. This product was not generated when beef heart mitochondrial aconitase was used as the source of iron. PMID- 8240339 TI - Defensin interferes with the activation of neutrophil NADPH oxidase in a cell free system. AB - HNP-1 [correction of NHP-1], a member of a family of microbicidal cationic, cystine-rich polypeptides abundant in the azurophilic granules of polymorphonuclear leukocytes (PMN) interfered with detergent-induced cell-free activation of superoxide-producing NADPH oxidase. IC50 of 3 microM has been evaluated for this dose dependent effect of defensin. Preactivated oxidase was not affected by HNP-1. The inhibitory effect could be relieved by raising the concentration of the cytosol or by the addition of a partially purified p47-phox. NADPH oxidase activated in the presence of HNP-1 exhibited an increased affinity for NADPH. PMID- 8240340 TI - Stimulation of calcium entry is prerequisite for DNA synthesis induced by platelet-derived growth factor in vascular smooth muscle cells. AB - The present study was conducted to determine a role of calcium entry in the growth promoting action of platelet-derived growth factor (PDGF) in cultured rat vascular smooth muscle cells. Addition of PDGF to quiescent cells induced a rapid elevation of cytoplasmic free calcium concentration, [Ca2+]c, followed by a sustained plateau of [Ca2+]c. Removal of extracellular calcium abolished the sustained phase, confirming that PDGF augments calcium entry. When calcium influx rate was determined by measuring initial uptake of [45Ca], PDGF induced a gradual increase in the rate of calcium entry. PDGF-mediated calcium entry became maximal after 90 min of the addition of PDGF, which persisted for at least 8 hrs. Calcium entry induced by PDGF was attenuated by nickel ion but not by a calcium channel blocker, nifedipine. When PDGF-induced calcium entry was blocked for a few hrs by either reducing extracellular calcium or by adding nickel, rate of entrance of S phase was reduced. In particular, VSMC stop progression when calcium entry is blocked in the middle of G1 phase. These results indicate that stimulation of calcium entry is prerequisite for initiation of DNA synthesis induced by PDGF in vascular smooth muscle cells. PMID- 8240341 TI - Membrane-associated N-myristoyltransferase activity is reduced in obese (fa/fa) Zucker rat liver. AB - N-Myristoyltransferase is the enzyme that catalyses the transfer of myristate from myristoyl-CoA to the NH2-terminal glycine residue of a number of protein of diverse functions. Many of the known myristoylated proteins are important in signal transduction. We have compared the activity of rat liver N myristoyltransferase from lean and obese (fa/fa) Zucker rats (a model for non insulin dependent diabetes mellitus, NIDDM). N-myristoyltransferase activity isolated from the particulate fraction of obese (fa/fa) Zucker rat liver was approximately 4.7-fold lower than the corresponding activity observed in either the controls or the vanadate-treated obese rat livers. This pattern was only observed in the particulate fraction; the homogenate and soluble N myristoyltransferase activities were not significantly different to the control activities. N-myristoyltransferase activity isolated from the brain of the four groups showed no significant variations. These results, and previous work [King, M. J., Pugazhenthi, S., Khandelwal, R. L. and Sharma, R. K. (1993) Biochim. Biophys. Acta. 1165, 259-262], would indicate that the rat liver particulate N myristoyltransferase activity appears to be inversely proportional to the level of plasma insulin, implicating insulin in the control of N-myristoylation. The specific activity of the particulate liver N-myristoyltransferase was approximately 10-fold higher than that of the soluble liver N myristoyltransferase, raising the possibility that N-myristoyltransferase exists in 2 populations, with the active form of N-myristoyltransferase residing in the membranous fraction. This situation could provide a system whereby N myristoylation is regulated by the translocation of N-myristoyltransferase from the cytosol to its active site in the membranes. PMID- 8240342 TI - Identification of two mPPAR related receptors and evidence for the existence of five subfamily members. AB - Two additional forms of mouse peroxisome proliferator activated receptor have been identified from cDNA libraries derived from mouse tissues using a low stringency cross-hybridization screening method. One is mNUCI, which has 97% amino acid sequence identity with hNUCI in its DNA binding domain. The other one is mPPAR gamma. The full-length cDNA of mPPAR gamma encodes a protein of 470 amino acids in length. The overall amino acid sequence identity is 75% as compared with xPPAR gamma, with the highest homology in the DNA binding region and ligand binding region, 97% and 86% identity, respectively. The discovery of two new forms of mPPAR and the recent cloning of hPPAR provide evidence for the existence of at least five different forms of receptor in the peroxisome proliferator activated receptor subfamily. PMID- 8240343 TI - Interleukin-4 modulates osteoclast differentiation and inhibits the formation of resorption pits in mouse osteoclast cultures. AB - Interleukin-4 (IL-4) is a well known lymphocyte growth factor, but it may also modulate the activity of other cell types. In the present study we show that IL-4 exerts a potent dose-dependent inhibitory effect on the resorptive activity of mouse bone cells, measured as the number of resorption pits (decreased down to 14% of control, p < 0.001) or the total resorbed area (down to 20% of control, p < 0.001). The results obtained in bone marrow and unfractionated bone cell cultures indicate that such an effect is mainly due to an inhibition of osteoclast precursors differentiation, rather than to a reduced activity of mature osteoclasts, and is not mediated by the IL-4-induced release of other soluble inhibitory factors. Likewise, although IL-4 may stimulate M-CSF expression, its inhibitory effect on osteoclast recruitment was not prevented by anti-M-CSF antibodies, suggesting that it was not mediated by M-CSF. These results point out IL-4 as an potentially important regulatory factor of osteoclast differentiation. PMID- 8240344 TI - Iron binding to human lactoferrin alters reactivity of the protein with plant lectins. AB - Binding of Fe by human apolactoferrin results in altered reactivity of the glycoprotein with plant lectins. Reaction with wheat germ agglutinin (WGA) and peanut agglutinin (PNA) was abolished with Fe binding. Reaction with the lectins from Datura stramonium (DSA) and Aleuria aurantia (AAA) was significantly reduced but not fully abolished on Fe binding, while reaction with the Artocarpus integrifolia lectin (Jacalin) and Sambucus nigrabark (SNA) was not changed at all. Loss of WGA reactivity occurred when only one of two Fe binding sites on the molecule was saturated. The results demonstrate conformational changes that are associated with high-avidity binding of Fe by lactoferrin. PMID- 8240345 TI - Identification of the heat shock protein of Neurospora crassa corresponding to the stress-inducible peroxidase. AB - Heat shock and other stress treatments, resulting in thermotolerance in Neurospora crassa cells, stimulate the induction of a peroxidase at a high level. The putative gene encoding this heat shock-induced peroxidase (hspp) has been cloned, using a cDNA clone of the manganese peroxidase of Phanerochaete chrysosporium, as a probe. Northern blot analysis of total RNA from heat-shocked cell showed the stress-dependent accumulation of a approximately 10 kb transcript. The identity of the HSP, corresponding to the heat-induced peroxidase, was established using hybrid-arrested in vitro translation. PMID- 8240346 TI - Evidence for a common binding site for omeprazole and N-ethylmaleimide in subunit A of chromaffin granule vacuolar-type H(+)-ATPase. AB - Vacuolar-type H(+)-ATPase from adrenal chromaffin granules was found to be sensitive to omeprazole, a known gastric H+/K(+)-ATPase inhibitor, the concentration required for 50% inhibition being 80 microM freshly-prepared and 12 microM acid-treated reagent. ATP and ADP protected the enzyme from inhibition by omeprazole. The activity of the inhibited enzyme was restored by the addition of reduced glutathione. Omeprazole protected the enzyme from inhibition by N ethylmaleimide and its binding to the subunit A. As subunit A has a nucleotide binding site(s) and as a cysteine residue is involved in the inhibition by N ethylmaleimide, these results suggested that the two sulfhydryl reagents bind to the same cysteine residue near the nucleotide binding domain in the subunit A, resulting in inactivation of vacuolar-type H(+)-ATPase. PMID- 8240347 TI - Conversion of nitroxyl (HNO) to nitric oxide (NO) in biological systems: the role of physiological oxidants and relevance to the biological activity of HNO. AB - Nitroxyl (HNO) and nitric oxide (NO) are chemically related compounds in that NO is the one-electron oxidation product of HNO. Previous studies from this laboratory indicated that HNO elicits pharmacological effects that are similar to those elicited by NO, namely, vascular smooth muscle relaxation and stimulation of cyclic GMP formation. The objective of the present study was to determine whether HNO could be converted to NO under physiological conditions and thereby account for the pharmacological actions of HNO. Utilizing the method of chemiluminescence detection, HNO was found to be readily converted to NO by a variety of ubiquitous biological oxidants including oxygen, superoxide dismutase, methemoglobin and flavins. The potency of HNO as a vasorelaxant using isolated rabbit aortic rings was markedly increased 30-fold by superoxide dismutase, whereas the potency of the NO-donor compound, S-nitroso-N-acetylpenicillamine (SNAP), was increased only 2-fold. These data indicate that the ready conversion of HNO to NO may account for the biological activity of HNO. Thus, HNO and HNO donor compounds represent good sources of NO. PMID- 8240348 TI - Resistance to methotrexate is associated with selective changes of alpha 2,6- and alpha 2,3-sialyltransferase activities toward N-acetyllactosaminic sequences in human colon cancer cell line HT-29. AB - In previous works we established that the alpha 2,6-sialyltransferase acting on N acetyllactosaminic sequences [alpha 2,6(N)ST, E.C. 2.4.99.1] behaves, in colonic cells, as an oncodevelopmentally regulated enzyme. Subpopulations of the human colon cancer cell line HT-29 adapted to grow in 10(-5) M methotrexate (MTX), permanently retain the ability to differentiate as mucus-secreting cells when kept confluent for extended periods of time [Lesuffleur et al. (1991) J. Cell Biol. 115, 1409-1418]. In this study we have compared the activities of five sialyltransferases acting on N- or O-linked chains of glycoproteins in parental HT-29 and in the 10(-5) M MTX-resistant variant. Both cell lines were studied during the exponential phase of growth as well as after a long period of postconfluent culture (28-30 days). Regardless the culture conditions, resistance to 10(-5) M MTX is associated with a virtual disappearance of alpha 2,6(N)ST activity. This change results in a dramatic reduction of the reactivity of cell membranes with the fluorescent lectin from Sambucus nigra, specific for alpha 2,6 sialylated structures. The activity of the alpha 2,3-sialyltransferase which acts on N-acetyllactosaminic sequences increases about two times in postconfluent cultures of 10(-5) M MTX-resistant cells, suggesting a close relationship with the differentiation degree. No significative changes were observed in the activity of other sialyltransferases. PMID- 8240349 TI - Natural occurrence of Nuc in the sera of autoimmune-prone MRL/lpr mice. AB - We previously established a clone of cells termed KML1-7 which produces a soluble factor that boosts anti-DNA antibody production both in vitro and in vivo across the H-2 barrier. By using the purified protein, termed nucleobindin (Nuc), we cloned cDNA and produced recombinant(r) Nuc in E.coli. Although the purified rNuc showed biological activities such as anti-DNA antibody boosting and DNA binding, there was no evidence that Nuc is really associated with autoimmune status in lupus-prone MRL/lpr mice. Here we report that identification of Nuc was successful from the sera of MRL/lpr mice, but not from those of the substrain MRL/n mice, which show no apparent autoimmune syndrome at the same age of MRL/lpr mice, by means of immunochemical as well as N-terminal amino-acid sequencing methods. PMID- 8240350 TI - Purification of the 120 kDa component of the human nuclear factor of activated T cells (NF-AT): reconstitution of binding activity to the cis-acting element of the GM-CSF and IL-2 promoter with AP-1. AB - The cis-acting element of the granulocyte-macrophage colony-stimulating factor (GM-CSF) promoter, CLE0, is required for stimulation dependent expression of the GM-CSF gene by phorbol ester (PMA) and calcium ionophore (A23187) in T cells. We recently obtained evidence that NF-CLE0 gamma, one of the CLE0-binding factors, is similar to the nuclear factor of activated T cells, NF-AT. In the present study, we show that the affinity-purified NF-AT from nuclear extracts of human Jurkat T cells stimulated with both PMA and A23187 bound strongly to the CLE0 element and formed a NF-CLE0 gamma complex. This DNA-protein complex was competitively inhibited by oligonucleotides containing NF-AT and AP-1 binding sites, suggesting that the CLE0 gamma complex is identical to NF-AT and contains AP-1 proteins. Here, one component of NF-AT with an apparent molecular mass of 120 kDa on SDS-polyacrylamide gel electrophoresis was purified to near homogeneity by Mono Q chromatography. The purified 120 kDa protein reconstitutes NF-AT binding in combination with recombinant cJun/cFos heterodimer. Furthermore, we demonstrate that binding of this 120 kDa protein to both the NF-AT and the CLE0 sequences can be reconstituted with the addition of affinity-purified Jurkat AP-1 proteins. These results indicate that NF-AT (NF-CLE0 gamma), which is composed of the 120 kDa nuclear protein and AP-1 proteins, regulates the stimulation-dependent expression of the GM-CSF gene as it does the IL-2 gene. PMID- 8240351 TI - Crystallization and atomic resolution X-ray diffraction analysis of antichymotrypsin variants. AB - Crystals of two recombinant antichymotrypsin (rACT) variants have been prepared: variant rACT-T345R crystallizes in space group P2(1) (a = 109.2 A, b = 79.4 A, c = 111.9 A, beta = 116.3 degrees, with 2 molecules in the asymmetric unit), and variant ACT' crystallizes in space group P2(1)22(1) (a = 69.7 A, b = 77.2 A, c = 83.8 A, with one molecule in the asymmetric unit). The latter variant is an engineered dimer having the P3-P3' hexapeptide sequence of the related serpin, alpha 1-proteinase inhibitor, substituted for the corresponding wild-type sequence. Crystals of each variant diffract to a limiting resolution of 2.5 A, which represents the best diffraction yet achieved for a crystalline, inhibitory serpin. The exceptional quality of ACT' crystals probably arises from favorable protein-protein interactions as well as a stabilizing disulfide crosslink engineered between the monomers. PMID- 8240352 TI - The insulin receptor substrate (IRS-1) is a PEST protein that is susceptible to calpain degradation in vitro. AB - The insulin receptor substrate 1 (IRS-1) contains at least 11 sequence motifs that are rich in proline (P), glutamic acid (E), serine (S), and threonine (T), i.e., PEST regions. Proteins with PEST regions turn over rapidly. IRS-1 is degraded rapidly in vivo upon exposure of 3T3-L1 adipocytes to insulin. The intracellular, calcium-dependent, neutral proteases known as calpains are one possible mechanism by which IRS-1 may be degraded. To begin to investigate this possibility, purified exogenous calpain was shown to degrade IRS-1 in cell-free extracts from basal and insulin-treated cells and rat recombinant IRS-1 in vitro. Only two proteolytic fragments could be detected. One had a mol wt of approximately 79 kDa, arising from the C-terminus end, and the second had a mol wt of approximately 90 kDa arising from near the N-terminus, possibly a product of the same cleavage event, since the mol wt of IRS-1 from insulin-treated cells was approximately 170 kDa. These results suggest that IRS-1 may serve as a substrate for calpain in vivo, accounting for its rapid degradation. PMID- 8240353 TI - CX-397, a novel recombinant hirudin analog having a hybrid sequence of hirudin variants-1 and -3. AB - To elucidate the differential roles of N- and C-terminal halves of hirudins in thrombin inhibition, we produced novel recombinant hirudin analogs, CX-397 and CX 397R, having a hybrid amino acid sequence of hirudin variants-1 (HV-1) and -3 (HV 3). CX-397 is composed of the N-terminal half of HV-1 (HV-11-36) combined with the C-terminal half of HV-3 (HV-337-66). CX-397R is the opposite combination. Their anti-thrombin activity was determined by a fluorogenic enzyme assay and compared with that of recombinant HV-1 (rHV-1) and rHV-3. The order of the magnitude of dissociation constants (Ki) of these four hirudin analogs in thrombin inhibition was as follows: CX-397R (0.294 pM) > rHV-1 (0.148 pM) > rHV-3 (0.0593 pM) > CX-397 (0.0433 pM), indicating that CX-397 is the strongest inhibitor among them. PMID- 8240354 TI - Genomic typing of hepatitis C viruses from Korean patients: implications of genome variation in the E2/NS1 region. AB - Comparative nucleotide sequence studies of a 2.4-kb cDNA fragment (nt -49 to 2361) of Korean-type hepatitis C virus (HCV) were carried out. The nt sequences of the Korean HCV isolates have stronger sequence homologies with type II HCV (Japanese HCV-BK) than with types I (HCV-1) or III (HCV-6). In addition, the net sequences of a part of the core region of ten different samples confirmed that prevalent HCV in Korea belongs to type II with the exception of one sample which belongs to type III. The nt sequence of a cDNA fragment of E2/NS1 region of HCV, from six different serum samples, which comprises the hypervariable regions-1 (HVR-1) and -2 (HVR-2), revealed sequence heterogeneity with type II HCV (75.4 to 80.7% nt homology and 75.5 to 79.7% amino acid homology). However, it was relatively well conserved among the Korean isolates, in fact, aa sequences of HVR 2 as well as HVR-1, converged into two groups. Also we found consensus sequences (T.V.G..AGRTT.G..SLE......K) in HVR-1. Thus, we propose that the conserved patterns in HVR-1 and -2 are characteristics of the six different Korean isolates. PMID- 8240355 TI - An E. coli gene emrD is involved in adaptation to low energy shock. AB - E. coli adapt to uncouplers in the presence of a non-fermentable carbon source. Adaptation is accompanied by the restoration of the proton motive force and ATP. A collection of uncoupler-sensitive Tn,lacZ,kan fusion strains was obtained by ampicillin enrichment in the presence of TSA. One of the fusion strains was induced by uncouplers. The fusion gene emrD was mapped to min. 83.1, cloned and sequenced. EmrD is a member of the major facilitator family of pmf-dependent translocases and is homologous to a number of bacterial multidrug resistance pumps. Resistance to some uncouplers including TTFB was not affected by emrD, and growth recovery to this uncoupler was very sluggish. It is suggested that EmrD is a new bacterial multidrug resistance pump that participates in a low energy shock adaptive response. PMID- 8240356 TI - Cytochrome c oxidase mutations in Leber hereditary optic neuropathy. AB - New mitochondrial DNA mutations were discovered in the cytochrome c oxidase subunit III gene in 8 independent Leber hereditary optic neuropathy probands. A mutation at nucleotide position 9438 was found in 5 probands, changed highly conserved glycine-78 to serine (G78S), and was not found in controls. A mutation at nucleotide position 9804 was found in 3 probands, changed highly conserved alanine-200 to threonine (A200T), and also was not found in controls. The 9438 mutation is readily detected by the loss of a Stu 1 restriction site and the 9804 mutation is detected by the gain of an Mae III restriction site. These mtDNA mutations may represent the first convincing examples of cytochrome c oxidase (Complex IV) mutations associated with a human disease. PMID- 8240357 TI - Purification from adult pig testicular P-450 and 17 alpha-hydroxylase activity of P-450 containing liposomal membranes. AB - A cytochrome P-450 from adult pig testicular microsomes was purified to a specific content of 12 nmol P-450/mg protein. P-450 has a minimum molecular weight of 46000 +/- 1000, as judged on sodium dodecyl sulfate polyacrylamide gel electrophoresis. Adult testicular P-450 is prepared in the low-spin form with an absorbance maximum at 417 nm. The substrate-induced difference spectrum with progesterone is a typical I difference spectrum. P-450 was incorporated into liposomal membranes composed of phosphatidylcholine, and 17 alpha-hydroxylase activity was shown to amount to 15.5 nmol product/min/nmol of P-450. Furthermore, testicular cytochrome b 5 did not increase the 17 alpha-hydroxylase activity, and the activity was largely inhibited by the addition of sodium cholate, Emulgen 913 and testicular lipid. PMID- 8240358 TI - An aminoterminally extended P1-protamine variant in the boar. AB - We have found a minor form of boar sperm P1-protamine with a six amino acid long N-terminal extension. When this sequence was compared with the recently published gene sequence of boar P1-protamine, and with DNA sequences obtained from the boars studied here, a perfect match was found with the 5'-region upstream of the ATG initiation codon of the authentic P1-protamine. Since no extra in-frame ATG initiation codons are present in the 5'-region of the gene, we conclude that the translation of boar P1-protamine variant starts with a non-AUG initiation codon, CUG, which precedes the codon of the first amino acid (serine) of the sequenced variant. PMID- 8240359 TI - Presence of 3-deoxyglucosone, a potent protein crosslinking intermediate of Maillard reaction, in diabetic serum. AB - 3-Deoxyglucosone, a potent protein crosslinking intermediate of the Maillard reaction, was first detected in diabetic serum using gas chromatography/mass spectrometry. Serum concentration of 3-deoxyglucosone was elevated in diabetic patients as compared with healthy subjects. More notably, diabetic patients with nephropathy showed higher serum concentration of 3-deoxyglucosone than those without nephropathy. The increased serum concentration of 3-deoxyglucosone in the diabetic patients suggests that 3-deoxyglucosone may be responsible for the development of diabetic complications such as diabetic nephropathy by promoting the formation of advanced glycation end products. PMID- 8240360 TI - Ha-ras stimulates the transplasma membrane oxidoreductase activity of C3H10T1/2 cells. AB - The ras family of oncogenes, as well as the transplasma membrane electron transport system, have been implicated in cellular growth control. Here we present evidence that the Ha-ras oncoprotein increases the activity of the transplasma membrane electron transport system in the mouse embryonic fibroblast cell line C3H10T1/2. Both cytochrome c and ferricyanide are reduced at a faster rate by C3H10T1/2 cells which are expressing the Ha-ras oncogene. In addition, the Ha-ras transformed cells extrude protons faster in response to stimulation by external oxidants than their normal counterparts. These results suggest that transformation events initiated by the expression of the Ha-ras oncoprotein, p21, in C3H10T1/2 cells have a controlling effect on the transplasma membrane electron transport system. PMID- 8240361 TI - p53 gene mutations in pontine gliomas of juvenile onset. AB - Using polymerase chain reaction-single strand polymorphism(PCR-SSCP) and nucleotide analyses, p53 gene mutation was examined in 13 pontine gliomas many of which were of juvenile onset. A total of 15 mutations were detected in 8 cases, of which 5 revealed multiple or tandem mutations. The mutations included 6 G:C A:T and 4 A:T-G:C transitions and 4 G:C-T:A and 1 A:T-T:A transversions. There was only 1 transition at the CpG site. Normal tissues of the same patients revealed no mutation, suggesting that these mutations were somatic, but not of germ line, in nature. The pattern of mutation characterized by frequent multiple or tandem occurrence, predominancy of transition at non-CpG sites and relatively frequent transversions suggested that pontine glioma might be related with some mutagenic or carcinogenic agents. PMID- 8240362 TI - Detection of poly (U) hybridization using azido modified poly (A) coated piezoelectric crystals. AB - A technique has been developed which uses AT-cut, 9 MHz piezoelectric crystals for the immobilization of single stranded nucleic acids and the subsequent hybridization reactions. This hybridization technique is based on detecting small changes in mass on the surface of a piezoelectric crystal which occur upon immobilizing azido containing probe nucleic acid on the surface and hybridizing complementary target nucleic acid to previously immobilized probe. The immobilized probe could be used for repeated hybridization tests. The hybridization assays were performed in solution and the frequency measurements were performed in the dry state. PMID- 8240363 TI - Transition state analogs as affinity labels for human DNA methyltransferases. AB - A new class of affinity labels has been developed for human DNA (cytosine-5) methyltransferases. These oligodeoxynucleotides contain 5-fluorodeoxycytidine at a mispair within the recognition motif of the human enzyme. They were not effectively recognized by bacterial methyltransferases. They can be viewed as analogs of the intermediates transiently produced by methyltransferases during catalysis. Affinity labelling patterns suggest that both the structurally induced activity and the methyl-directed activity of the human enzymes operate by the same mechanism and reside on the same polypeptide chains. PMID- 8240364 TI - Intracellular magnesium homeostasis is involved in the functional recovery of preconditioned rat heart. AB - The role of ionic derangements in preconditioned hearts (PC) was assessed by measuring pHi, high-energy phosphates (HEP) and [Mg2+] by 31P NMR. Control (C) Langendorff rat hearts were subjected to 30' ischemia and 30' reflow. PC underwent 4x(2' ischemia + 3' reperfusion) before prolonged ischemia and reflow. In this model, the contractile recovery of PC hearts at the end of reflow (rate pressure product RPP: 50 +/- 12% vs. 5 +/- 5% of C, p = 0.004) is not related to higher HEP or pHi levels than in C. [Mg2+]i increased significantly during ischemia both in C and PC organs; upon reperfusion its level remained significantly high in C (p < 0.001), while it regained the normal value in PC hearts. This behavior might improve in turn the SR calcium handling in PC organs, eventually contributing to the contractile recovery. PMID- 8240365 TI - Comparison of bacteriorhodopsin/phospholipid interactions in DMPC and DMPG bilayers: an electron spin resonance spectroscopy and freeze-fracture electron microscopy study. AB - Bacteriorhodopsin (BR), separated from the Halobacterium halobium purple membrane (PM) lipids, broadened the main gel-fluid phase transition (Tc) of 1,2 dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers and 1,2-dimyristoyl-sn glycero-3-phosphoryl-3'-rac-glycerol (DMPG) bilayers to similar degrees compared to the respective protein-free bilayers. Analysis of two component ESR spectra from spin-labelled phospholipids suggested that, at temperatures above Tc, each 26 kDa BR was associated with 18-21 boundary lipids in both DMPC and DMPG bilayers. This was consistent with the random dispersive properties of the BR particles observed by freeze-fracture electron microscopy in DMPG and DMPC bilayer complexes when quenched from temperatures above Tc. DMPG, as shown for DMPC in other studies, did not aggregate BR into the 2D-hexagonal arrays of PM at temperatures below Tc. Despite the presence of positively-charged amino acid residues which project from the BR into the polar headgroup region of the phospholipid bilayer, BR exhibited a lower affinity for the negatively-charged phosphoglycerol headgroup compared to the zwitterionic phosphocholine headgroup, suggesting that ionic interactions between BR and phospholipid headgroups are not significant and serve a passive function, perhaps facilitating the close packing of BR into 2D-arrays. PMID- 8240366 TI - Metabolism of hexadecanol by rat type II pneumonocytes. AB - Type II pneumonocytes isolated from adult rats incorporate radioactivity from 1 [3H]hexadecanol into either lipids. The radioactivity is distributed between the neutral lipids, phosphatidylcholines, and phosphatidylethanolamines. The neutral lipid fraction is almost exclusively composed of triradylglycerols. The potent ether-containing glycerophospholipid, platelet-activating factor, has been shown to stimulate glycogen breakdown and surfactant secretion. In consideration that a long chain alcohol (hexadecanol) is added to certain artificial surfactant preparations which are utilized in the treatment of respiratory distress syndrome of the newborn, the metabolism of hexadecanol by the type II pneumonocytes should be taken into account. PMID- 8240367 TI - An inborn error in epidermal growth factor prohormone metabolism in a mouse model of autosomal recessive polycystic kidney disease. AB - The C57BL/6J CPK heterozygous breeders secrete in urine a variant EGF-prohormone with a molecular mass of 154 kDa in addition to the normal 165 kDa EGF prohormone. The 154 kDa prohormone is secreted as a heterodimer with the normal 165 kDa prohormone. The phenotypically normal littermates, like their parents, secrete the 154 and 165 kDa EGF-prohormones in urine while their cystic siblings secrete neither protein. Examination of renal extracts of normal littermates revealed the presence of the 165 kDa but not the 154 kDa EGF-prohormone; renal extracts of cystic siblings contained neither protein. Cyst fluid, however, contained 56 and 49 kDa EGF-immunoreactive proteins in high concentrations. The data suggest that in the absence of normal 165 kDa prohormone, the 154 kDa EGF prohormone undergoes proteolysis and that the resultant fragments function as cystogens. Since normal siblings do not acquire renal cystic disease despite expressing the variant 154 kDa EGF-prohormone while the affected littermates, which lack the normal 165 kDa EGF-prohormone, manifest renal cystic disease, we suggest that congenital polycystic kidney disease is due to an inborn defect in the synthesis and secretion of the normal 165 kDa renal EGF-prohormone. PMID- 8240368 TI - Labeling of the active site of alpha-chymotrypsin with N-nitroso-N benzylacetamide and related compounds. AB - alpha-Chymotrypsin (CT) is slowly inhibited by N-nitroso-N-benzylacetamide (1) at 25 degrees C. The 13C-NMR spectrum of the hydrolysate of 13C-enriched 1-inhibited CT shows five new signals at 52.33, 44.55, 43.72, 36.79, and 32.90 ppm resulting from alkylation of the side chains and also of amide linkages of the enzyme backbone by benzyl carbocations produced in the active site. The alkylation pattern is different from those observed in the inhibitions of CT with D-N nitroso-N-[alpha-13C]-benzyl-N'- isobutyrylalaninamide (2a) and D-N-nitroso-N [alpha-13-C]-benzyl-N'-isobutyrylphenylalaninamide (2b) (White et al. JACS 1990, 112, 1956-1961). The chemical shift data suggest that the 52.33 ppm signal stems from N-benzylglycine, the 36.79 ppm signal from S-benzylcysteine, and the 32.90 ppm signal from 2-benzyltryptophan. A synthesis of the latter compound was developed. PMID- 8240369 TI - Role of calcium ion in induction of apoptosis by etoposide in human leukemia HL 60 cells. AB - The epipodophyllotoxin derivative etoposide, an inhibitor of DNA topoisomerase II, has been found to induce internucleosomal DNA fragmentation, characteristic of apoptosis. In the present study, we examined the effect of Ca2+ depletion on VP-16-induced endonucleolytic DNA cleavage in HL-60 cells. VP-16-induced internucleosomal DNA fragmentation could not be blocked by the extracellular Ca2+ chelator EGTA. However, an intracellular Ca2+ chelator BAPTA-AM, which was added after treatment with VP-16, abolished both internucleosomal DNA fragmentation and the morphologic features of apoptosis. No significant increase of intracellular Ca2+ was found after VP-16 treatment. We demonstrate for the first time that preexisting intracellular Ca2+ plays an essential role in induction of apoptosis by VP-16 in HL-60 cells. PMID- 8240370 TI - Lupus autoantigen Ku protein binds HIV-1 TAR RNA in vitro. AB - The control of HIV-1 gene expression depends upon interaction of Tat with the trans-activation responsive (TAR) element present at the 5' end of all HIV-1 transcripts. The TAR sequence forms a stable hair-pin structure that binds Tat and several cellular factors in vitro. In the absence of Tat, TAR acts as a transcription terminator. Tat in conjunction with a cellular factor(s) acts to increase the elongation capacity of the transcription complex. Here we report that Ku protein, the autoantigen in patients with systemic lupus erythematosus, binds TAR RNA in vitro with high affinity and specificity forming a single protein-RNA complex. Ku, a heterodimer of Ku72 and Ku86 that non-specifically binds the ends of DNA fragments, appears to recognize the terminal loop of TAR RNA. UV-crosslinking showed that both subunits of Ku are in proximity to the RNA. Further, Ku shows a 5-fold higher affinity for TAR RNA than for the ends of dsDNA. As Ku is involved in the stimulation of the elongation property of the RNA polymerase II and activation of several transcription factors, the specific interaction of Ku with TAR raises intriguing possibilities for its function in HIV-1 gene expression. PMID- 8240371 TI - A novel approach for isolation and mapping of intron mutations in a ribonucleotide reductase encoding gene (nrdB) of bacteriophage T4 using the white halo plaque phenotype. AB - The nrdB gene of bacteriophage T4 codes for the small subunit of ribonucleotide reductase and contains a 598 base pair self splicing intron which is closely related to other group I introns of T4 and eukaryotes. The screening, isolation and mapping of the nrbB intron mutations was conducted by the strategic usage of the white halo phenotype exhibited by T4 mutants defective in dhydrofolate reductase or thymidylate synthase. We have isolated 159 hydroxylamine-induced nrdB mutants, determined which mutations are in nrdB by marker rescue with clones of the nrdB gene and have mapped these mutations by marker rescue using subclones of the nrdB intron. Thirty out of the 159 nrdB mutations are in or near the intron. These mutations cluster towards the ends, mainly the 3' end. We have performed deletion mapping to further map mutations in the 3' end of the intron. The mutations map in regions of conserved structural elements, thus supporting secondary structure predictions similar to those of the well studied td intron in the T4 gene coding for thymidylate synthase. PMID- 8240372 TI - Autosomal dominant hypoalphalipoproteinemia due to a completely defective apolipoprotein A-I gene. AB - Primary hypoalphalipoproteinemia is associated with atherosclerosis and exhibits significant familial aggregation. To reveal the presence of autosomal dominant hypoalphalipoproteinemia due to a completely defective apolipoprotein A-I gene, the apolipoprotein A-I gene was analyzed in a Japanese family with low levels of HDL cholesterol and apolipoprotein A-I. An insertion of a C in the region of the seven C run between codons 3 and 5 was detected in the apolipoprotein A-I gene. The heterozygous state for the mutation was associated with approximately 50% of the normal HDL cholesterol levels and of the normal apolipoprotein A-I levels. The data suggest that a part of familial hypoalphalipoproteinemia might be an autosomal dominant trait due to a completely defective apolipoprotein A-I gene. PMID- 8240373 TI - Detection of K-ras codon 12 mutation by polymerase chain reaction and differential dot-blot hybridization in sodium saccharin-treated human RSA cell. AB - Using polymerase chain reaction and differential dot-blot hybridization, point mutations of K-ras codon 12 were investigated in hyper-mutable human RSa cells treated with sodium saccharin. Cells carried mutational changes after 6 days of treatment at sodium saccharin concentrations of 15 to 30 mg/ml. Results imply that sodium saccharin is a mutagen which induces genetic mutation and, furthermore, that the assay system applied here will be beneficial in identifying mutagen-induced K-ras oncogene mutations in human cells in vitro. PMID- 8240374 TI - Angiotensin II regulates human vascular smooth muscle alpha-actin gene expression. AB - Angiotensin II (AII) has been shown to induce vascular smooth muscle cell (VSMC) hypertrophy and increased expression of vascular cytoskeletal proteins. We have studied basal and AII-induced expression of the chloramphenicol acetyl transferase (CAT) gene driven by three fragments of the human vascular smooth muscle (SM) alpha-actin promoter. We show basal CAT expression driven by the three fragments of the promoter when the constructs are transiently transfected into rat aortic VSMCs. AII in a concentration-dependent manner (1.0 nM to 10 microM) increased expression of the CAT gene driven by 896 bp fragment. When comparing the 896 bp fragment to fragments successively deleted at the 5' end (674 bp and 258 bp respectively), AII markedly stimulated CAT expression driven by the 896 bp fragment (257 +/- 31% over control, p < 0.01), stimulated CAT expression driven by 674 bp fragment to an apparently lesser degree (189 +/- 20% over control, p < 0.01), and tended to stimulate CAT expression driven by the 258 bp fragment, though not significantly greater than baseline (157 +/- 28% of control). These data suggest that AII exerts transcriptional regulation of human SM alpha-actin gene through activation of cis-acting element(s) in an upstream area localized between positions -258 and -896 of the SM alpha-actin promoter. Such findings help establish the role of AII in enhancement of expression of components of the contractile apparatus. PMID- 8240375 TI - Angiotensin converting enzyme inhibitors or DuP753 prevent neointimal formation following balloon injury with single topical or multiple systemic application. AB - Angiotensin II plays an important role in neointimal formation after vascular injury. Our objectives were 1) to investigate the difference between angiotensin converting enzyme inhibitors (captopril, delapril) and an angiotensin II subtype 1 (AT1) receptor antagonist (DuP753) in suppressing neointimal proliferation; and 2) to investigate the antiproliferative effects of these drugs given topically to the injured vessels. All these treatments effectively prevented neointimal formation (p < 0.01). Even a single topical application of either type of drug with F127 pluronic gel to be injured vessel after ballooning is found to be significantly effective probably due to the inhibition of smooth muscle cell migration (p < 0.01). Multiple systemic application of angiotensin converting enzyme inhibitors was more effective than that of DuP753 at the same blood pressure level. The effectiveness of topical application of these drugs suggests clinical usefulness after angioplasty or vascular surgery. PMID- 8240376 TI - T-jump fluorescence relaxation study of the binding of ethidium cation to natural DNA. AB - T-jump chemical relaxation experiments with fluorescence detection were carried out on the binding of ethidium cation to calf thymus DNA over the temperature range from 11 to 35 degrees C for ethidium concentrations from 500 nM to 2 microM, DNA concentrations from 7.5 to 60 microM (base pairs), and KCl concentrations from 20 to 500 mM. In contrast to the findings of earlier investigations, no salt or DNA concentration or temperature dependences were seen that would support a mechanism of intercalation directly from solution or from an electrostatically bound exterior site in rapid equilibrium with solution. It is suggested that the rate-determining process of intercalation in vivo involves large scale dynamics of the DNA itself. PMID- 8240377 TI - Expression of receptors for advanced glycosylation end products on renal cell carcinoma cells in vitro. AB - Proteins that have been modified by long-term expose to glucose accumulate advanced glycosylation end products (AGEs) as a function of protein age. In these studies, we have examined the interaction of AGE-protein with renal cell carcinoma cells (RCC) in vitro, using AGE-modified bovine serum albumin (AGE-BSA) as a probe. AGE-BSA showed tendency to induce in vitro cell growth of RCC cells and promoted the production of interleukin-6 (IL-6), an in vitro autocrine growth factor. Reverse transcriptase-polymerase chain reaction analysis revealed that RCC cells used here express mRNA for a receptor for AGEs (RAGE). These results suggested that AGEs taken up through RAGE on RCC cells might play a role in promoting the growth of RCC cells. PMID- 8240378 TI - Inhibition of restriction endonuclease cleavage site via triple helix formation by homopyrimidine phosphorothioate oligonucleotides. AB - The ability of pyrimidine rich oligonucleotide phosphorothioate to form stable triple helical structures with the sequence containing the recognition site for the class II-S restriction enzyme Ksp 632-I was examined. First, we synthesized double strand oligonucleotides corresponding to the SV40 sites and studied their interaction with homopyrimidine oligodeoxyribonucleotides including replacement of the other chain either PS group (SO-ODNs) in second nucleotide position (from 5'-terminus) and end capped with the PS group at both 3'- and 5'-ends (S2O-ODNs). The resulting perfect DNA triplexes were detected by gel-mobility shift. The phosphorothioate oligonucleotide analogues (SO-ODNs) and (S2O-ODNs) were shown to inhibit enzymatic cleavage under conditions that allow for triple helix formation. Inhibition is sequence-specific and occurs in the micromolar concentration range. Of particular interest is the Sp-phosphorothioate analogue (Sp-SO-ODNs) which inhibited endonuclease more than the other phosphorothioate oligonucleotide analogues (Rp-SO-ODNs or S2O-ODNs). PMID- 8240379 TI - The radiochemical assay for monoamine oxidase activity. Problems and pitfalls. PMID- 8240380 TI - Effects of tannic acid on 12-O-tetradecanoylphorbol-13-acetate-induced protein kinase C activation in NIH 3T3 cells. AB - Tannic acid (TA) is a naturally occurring phenol, which has been found to display an antipromotion effect on mouse skin carcinogenesis. In order to explore the molecular mechanism, we have examined the process of 12-O-tetradecanoylphorbol-13 acetate (TPA)-induced protein kinase C (PKC) activation, including phorbol ester binding, enzyme translocation, autophosphorylation and substrate phosphorylation, and finally the TPA-stimulated DNA synthesis. In an initial study, we found that TA slightly inhibited [3H]phorbol dibutyrate (PDBu) binding to intact cells, and only 30% reduction of phorbol ester binding was observed at the highest dose of TA (100 microM). Further analysis by Scatchard plot showed that TA reduced the Bmax of [3H]PDBu from 1.3 pmol/10(6) cells to 1.1 pmol/10(6) cells, but the Kd was increased from 24 to 30 nM. Analysis by western blot indicated that TA did not interfere with the TPA-induced PKC translocation, whereas TA effectively blocked the TPA-evoked phosphorylation of the membrane-bound PKC moiety and its 80 kDa substrate in a dose-dependent manner. We also found that pre- or post treatment with TA both lead to a similar reduction of 80 kDa protein phosphorylation and that the TPA-stimulated DNA synthesis was also inhibited by TA in a dose-dependent manner, suggesting that the blockage of protein phosphorylation by TA was of biological significance. In conclusion, the work presented here demonstrated that the antitumor promoting effect of TA was not mediated by competing for the binding site with phorbol ester or interrupting the PKC translocation, but rather by effectively blocking phosphorylation by membrane bound PKC, possibly through altering the biophysical properties of the membrane environment. PMID- 8240381 TI - Ascorbic acid uptake by isolated rat hepatocytes. Stimulatory effect of diquat, a redox cycling compound. AB - The toxicity of redox cycling compounds which generate the formation of active oxygen species is commonly accepted to be associated with a decrease of cellular reductants involved in cellular defence. However, when hepatocytes were incubated with diquat, an established redox cycler, in the presence of ascorbic acid (AA) (1 mM), the intracellular level of AA was increased. The effects of diquat on AA uptake were investigated in isolated rat hepatocytes. Incubation of hepatocytes with diquat plus AA (1 mM) resulted in about a 2-fold increased accumulation which occurred in a time-dependent manner reaching a steady state after 15 min at 37 degrees. The initial AA uptake rate was dependent on the AA concentration added. This process is described by Michaelis-Menten kinetics (apparent Km = 953 +/- 59 microM and Vmax = 2.68 nmol/min/10(6) cells). Characterization of AA accumulation showed it to be inhibited: by incubation at 4 degrees; with carbonyl cyanide p-trifluoromethoxyphenylhydrazone, an inhibitor of intracellular ATP production; by decreasing the extracellular Na+ concentration or incubating with ouabain; with pfloretin, a glucose transport inhibitor; and with glucose, a competitive inhibitor of AA transport. Replacement of AA with its oxidized form, dehydroascorbic acid, in the absence of diquat enhanced AA accumulation by 2.5 fold and apparently prevented further accumulation by added diquat. In addition, maintaining AA reduced with dithiothreitol inhibited the diquat effect. Diquat induced AA accumulation was inhibited (65%) by desferrioxamine, a free-iron chelator, but not by catalase and/or superoxide dismutase or different antioxidants. In contrast, incubation with other active oxygen species generating systems including bipyridilium structural analogues, paraquat and benzyl viologen, had no effect on AA accumulation in hepatocytes. These results suggest that diquat-induced AA accumulation by hepatocytes occurs by a specific mediated transport system rather than as a consequence of cytotoxicity and may involve the presence of free-iron. PMID- 8240382 TI - Compartmentation of intestinal drug sulphoconjugation. Incorporation of luminal and contraluminal [35S]sulphate into 1-naphthol by the isolated mucosa of guinea pig jejunum and colon. AB - Compartmentation of 1-naphthol metabolism was inferred from the metabolite pattern and distribution in the isolated mucosa of guinea pig intestine mounted in a flux chamber (Sund and Lauterbach, Arch Pharmacol Toxicol 58: 74-83, 1986). To verify the existence of these compartments the dependence of [35S]sulphate incorporation into 1-naphthol sulphate on the side of administration of 1 naphthol and [35S]sulphate was determined. Isolated mucosae were pre-equilibrated with [35S]-sulphate (5 x 10(6) cpm/mumol, 1 mM) for 30 min and subsequently incubated for 15 min with 50 microM 1-naphthol. The three 1-naphthol sulphate fractions (luminal side, blood side and tissue) were assayed by HPLC and liquid scintillation counting; their specific activity was calculated as percentage of the specific activity of the inorganic sulphate administered. 1-Naphthol glucuronide was also measured. In jejunal experiments: after luminal administration of 1-naphthol, 1-naphthol sulphate appeared almost exclusively in the luminal solution; its specific activity approached 70% and 3%, when [35S]sulphate was added to the luminal and blood side, respectively. After introducing 1-naphthol and [35S]sulphate on the blood side, a high and similar specific activity of 50-60% was observed in all three 1-naphthol sulphate fractions (luminal and blood side, tissue) though adding [35S]sulphate to the lumen side decreased the specific activity to 10-20%. In experiments on colonic mucosa: a specific activity both of luminal and blood side 1-naphthol sulphate of more than 50% was observed with blood side [35S]sulphate irrespective of the side of 1-naphthol administration. When [35S]sulphate was placed on the luminal side the specific activity of blood side 1-naphthol sulphate dropped to only 3%, and that of luminal 1-naphthol sulphate ranged between 6% and 20%. Supplementary experiments in which mucosae were exposed to 1-naphthol and [35S]sulphate for 45 min without preincubation showed a tendency to decrease the lumen: blood distribution ratio of 1-naphthol sulphate. However, the general pattern of 1 naphthol sulphate specific activity remained unchanged. The experiments provide further evidence that the jejunal conjugation of phenolic drugs is being performed in two major compartments, which are accessible from the lumen ("luminal compartment") and blood ("systemic compartment") side. The luminal compartment seems practically inaccessible to blood side sulphate as is the systemic compartment for luminal 1-naphthol. In the colonic mucosa, a major "systemic compartment" receiving its sulphate from the blood side is the site for most of the events, but a minor "luminal compartment" seems to be involved as well. PMID- 8240383 TI - Vitamin K2 (menatetrenone) inhibits prostaglandin synthesis in cultured human osteoblast-like periosteal cells by inhibiting prostaglandin H synthase activity. AB - Prostaglandin (PG) E2, a potent bone-resorbing agent, is synthesized in osteoblast-like cells. Since vitamin K reportedly plays an important role in bone metabolism, we investigated the effects of vitamin K2 (menatetrenone) on PGE2 production by human osteoblast-like periosteal cells. In cells incubated with menatetrenone (1 microgram/mL = 2.25 x 10(-6) M) for 2 days, PGE2 production was reduced to 50% of that in untreated control cells. This inhibition was dose and time dependent for up to 10 micrograms/mL and 20 days, respectively, and involved two major steps. In one of these menatetrenone at doses of 0.5-10 micrograms/mL dose dependently inhibited the calcium ionophore A23187-induced release of arachidonic acid (AA) from membrane phospholipids, and in the other the conversion of AA to PG was inhibited, as evidenced by the PG-synthesizing activity in the homogenates of menatetrenone-treated cells with AA being lower than that in untreated cells. The inhibitory effect was almost identical to that for PG production. The PG synthesizing activity in cell homogenates was inhibited only by a high concentration of menatetrenone (10 micrograms/mL) when this was added directly. Menatetrenone (1 microgram/mL) also inhibited 52% of the purified PGH synthase activity from a ram seminal vesicle. This study shows that menatetrenone inhibited PGE2 release from cells by inhibiting both PG production steps, AA release from the membrane and PG synthesizing activity with AA. Inhibition of PGE2 production by menatetrenone might be important in improving bone metabolism. PMID- 8240384 TI - Reduction of dapsone hydroxylamine to dapsone during methaemoglobin formation in human erythrocytes in vitro--II. Movement of dapsone across a semipermeable membrane into erythrocytes and plasma. AB - We have used an in vitro two-compartment model, to investigate the ability of dapsone, formed by erythrocyte-mediated detoxification of its hydroxylamine metabolite, to escape the cells and cross a semi-permeable membrane into both plasma and other erythrocytes. Both diethyl dithiocarbamate (DDC) treated and untreated erythrocytes were incubated with dapsone hydroxylamine and dialysed against either fresh cells or plasma. Methaemoglobin was predominantly detectable in compartment A although the presence of low levels of methaemoglobin in compartment B indicated that the hydroxylamine itself had crossed the membrane. In contrast to methaemoglobin disposition, recovery of dapsone was higher (P < 0.05) in compartment B compared with A for all three treatment groups at 30 and 60 min, but not at the remaining time points. Regression analysis of the cumulative recovery of dapsone over 150 min in all three treatment groups for both compartments A and B showed correlation coefficients close to unity. In compartment A, analysis of the mean slopes of the regression lines indicated that, overall, significantly more dapsone was recovered from group 1 (erythrocytes, hydroxylamine and DDC dialysed against untreated red cells) compared with group 3 (erythrocytes and hydroxylamine dialysed against plasma) (0.22 +/- 0.05 vs 0.09 +/- 0.005; P < 0.025). Also in compartment A, significantly more dapsone was recovered from group 2 (erythrocytes and hydroxylamine dialysed against untreated red cells) compared with group 3 (erythrocytes and hydroxylamine dialysed against plasma: 0.16 +/- 0.02 vs 0.09 +/ 0.005). In compartment B, dapsone recovery was significantly greater in group 1 (erythrocytes, hydroxylamine and DDC dialysed against untreated red cells; slope of regression line: 0.59 +/- 0.05) compared with group 2 (erythrocytes and hydroxylamine dialysed against untreated red cells; slope of line: 0.28 +/- 0.02, P < 0.005). In addition, dapsone recovery was significantly greater in group 1 (0.59 +/- 0.05) compared with group 3 (erythrocytes and hydroxylamine dialysed against plasma; 0.21 +/- 0.02, P < 0.005). Dialysis of erythrocytes with dapsone itself over 120 min caused no detectable methaemoglobin formation. The process of erythrocyte-mediated dapsone formation from its hydroxylamine may feasibly occur in vivo and contribute to the systemic persistence and therapeutic effect of dapsone. PMID- 8240385 TI - Histamine lipolytic activity and semicarbazide-sensitive amine oxidase (SSAO) of rat white adipose tissue (WAT). AB - Histamine has previously been described as a possible substrate for the semicarbazide-sensitive amine oxidase activity (SSAO) of rat white adipose tissue (WAT). We report here on a histamine function in this tissue which concerns the activity of this deaminating system distinct from the classical diamine oxidase. Our results show that: (1) histamine plays a role in controlling rat adipose tissue lipolysis with the contribution of H1 and H2 receptors that participate in histamine lipolysis in an opposite way. Both H1 and H2 roles can be differentiated using selective agonists (2- and 4-methyl histamine) and antagonists (pyrilamine and cimetidine); (2) histamine might also control rat lipolysis induced by noradrenergic agonists; (3) the SSAO present in rat WAT controls histamine levels at the receptor sites as shown by the modification of histamine lipolytic potency obtained when inhibitors of this enzyme are used. PMID- 8240386 TI - Enhanced formation of reactive species from cis-diammine-(1,1 cyclobutanedicarboxylato)-platinum(II) (carboplatin) in the presence of oxygen free radicals. AB - Experiments were designed to investigate the influence of oxygen free radicals on the rate of conversion of the anticancer drug cis-diammine-(1,1 cyclobutanedicarboxylato)platinum(II) (CBDCA) to reactive species able to bind to DNA. A system containing the Fe-EDTA chelate and ascorbate was used to generate free radicals. The rate of drug conversion to by-products, during incubation in chloride-free phosphate buffer at 37 degrees, was determined by HPLC analysis and found to be approximately 10 times faster in the presence of the free radical generating system, compared to CBDCA alone. The hydroxyl radical scavenger, mannitol, was able to reduce the rate of CBDCA conversion significantly, while an enhancing effect was observed in the presence of superoxide dismutase. The platinum containing species, which are formed in the presence of free radicals, were demonstrated to react with isolated salmon sperm DNA. The rate of platinum binding to DNA during incubation of CBDCA in the presence of the Fe EDTA/ascorbate system was markedly enhanced. No effect on platinum binding to DNA during incubation with cis-diamminedichloroplatinum(II) (CDDP) in the same experimental conditions was observed, thus excluding an increased susceptibility of DNA itself to binding of platinum, due to DNA damage induced by free radicals. These findings support the hypothesis that the increased conversion of CBDCA, previously observed in our laboratory, which occurs in the presence of hemoglobin could be mediated by a Fenton-like reaction resulting in oxygen free radical production, thus providing potential clues to improvements in the clinical use of this drug. PMID- 8240387 TI - Hexachlorobenzene-induced hypothyroidism. Involvement of different mechanisms by parent compound and metabolite. AB - Rats received repeated oral treatment with different doses of hexachlorobenzene (HCB) (0-3.5 mmol/kg) for 2 or 4 weeks. Measurements of thyroid hormone status after 2 weeks showed a dose-dependent decrease of total thyroxine (TT4) levels, decreased free thyroxine (FT4) levels and little change of total triiodothyronine (TT3) levels. The effects on thyroid hormone status were more pronounced after 4 weeks and also included increased thyroid stimulating hormone (TSH) levels. These conditions suggest that HCB had induced hypothyroidism in these animals. Indications for occupation of thyroid hormone binding proteins were found in serum of exposed animals. The major metabolite pentachlorophenol (PCP) also caused, by competitive interactions with thyroid hormone binding proteins in serum, a rapid and dose-dependent decrease of TT4 and FT4 levels, but not of TT3 levels in serum. The decrease of serum TT4 levels by repeated dosing with 3.5 mmol HCB/kg for 4 weeks could be attributed to competitive interactions of PCP with hormone serum binding proteins and to increased metabolism induced by HCB to an equal degree. At lower dose levels or with shorter dosing periods, increased metabolism of T4 is the main cause of decreased TT4 serum levels. This is the first indication that a similar effect is caused simultaneously by the parent compound and its metabolite through different and independent mechanisms. PMID- 8240388 TI - Phase I and II biotransformations in living CaCo 2 cells cultivated under serum free conditions. Selective apical excretion of reaction products. AB - CaCo 2 cells, cultivated in a synthetic, serum-free nutritive medium on poly (ethylene terephthalate) membranes, form a confluent monolayer of differentiated cells, with the apical and basolateral poles exposed to the upper and lower compartments, respectively, of bicameral culture inserts (Halleux and Schneider, In Vitro Cell Dev Biol, 27A: 293-302, 1991). This cell culture system allows the passage of intact mannitol by the paracellular route and the transcellular diffusion of testosterone which appears mainly as a biotransformed unconjugated metabolite. When ethoxyresorufin is added to either the apical or basolateral poles of living CaCo 2 cells, resorufin is formed, and more than 80% is excreted at the apical pole. Under our experimental conditions, no detectable amounts of glucurono- or sulfconjugates are found. Methylcholanthrene and phenobarbital increase the biotransformation of ethoxyresorufin 50 and 3 times, respectively, and induce that of benzoxyresorufin, but not of pentoxyresorufin which remains absent under all conditions. These substances do not affect the proportion of resorufin recovered at the apical role. Verapamil inhibits by 25% the release of resorufin but does not affect its distribution. Chlorodinitrobenzene is conjugated with glutathione and at least two-thirds of the product is excreted at the apical pole; methylcholanthrene and phenobarbital do not increase this activity. These results demonstrate that differentiated CaCo 2 cells, under serum free conditions, perform phase I and II reactions and that the biotransformation products are selectively excreted at the apical pole. PMID- 8240389 TI - The antileukemic alkaloid fagaronine is an inhibitor of DNA topoisomerases I and II. AB - The antileukemic alkaloid, fagaronine, is a potent differentiation inducer of various hematopoietic cell lines. We show here that fagaronine is a DNA base-pair intercalator with a K(app) of 2.1 x 10(5) M-1 for calf thymus DNA. Fagaronine inhibits the catalytic activity of purified calf thymus topoisomerase I as shown by relaxation of supercoiled plasmid DNA followed by electrophoresis in neutral as well as in chloroquine-containing gels. The catalytic activity of topoisomerase I is inhibited at concentrations above 30 microM. Fagaronine also inhibits the catalytic activity of purified calf thymus topoisomerase II at concentrations above 25 microM as shown by decatenation of kinetoplast DNA. Fagaronine stabilizes the covalent DNA-enzyme reaction intermediate (the cleavable complex) between topoisomerase I and linear pBR322 DNA at concentrations up to 1 microM. Further increase of the fagaronine concentration leads to a progressive decrease in the cleavable complex formation, which is totally inhibited at 100 microM. In contrast, up to 1 microM fagaronine has no effect on cleavable complex formation between purified calf thymus topoisomerase II and linear pBR322 DNA, whereas cleavable complex formation is inhibited at higher concentrations. Exposure to fagaronine results in an increase in DNA protein complex formation in intact P388 murine leukemia cells. P388CPT5 cells, which have an altered topoisomerase I activity, are 4-fold resistant to the growth inhibitory effects of fagaronine compared to the parental cell line. Similarly, DC-3F/9-OH-E Chinese hamster fibrosarcoma cells, which have an altered topoisomerase II activity, are about 5-fold resistant to the growth inhibitory effects of fagaronine. We conclude that fagaronine is an inhibitor of both DNA topoisomerase I and II and propose that this might play a role in the cytotoxic activity. PMID- 8240390 TI - Hydrolysis and binding of a toxic stereoisomer of soman in plasma and tissue homogenates from rat, guinea pig and marmoset, and in human plasma. AB - The fallen concentration of one of the two isomers of soman (1,2,2 trimethylpropyl methylphosphonofluoridate), i.e., C(+)P(-)-soman, was investigated in plasma and in homogenates of brain, lung, liver, kidney, diaphragm, skeletal muscle and mucosa of small intestines from rat, guinea pig and marmoset, and in human plasma (pH 7.5, 37 degrees). The decrease of the isomer concentration was followed by gas chromatographic determination of the residual concentration and proceeded in two phases due to a very rapid saturation of covalent binding sites for the isomer followed by catalysed hydrolysis. Estimates for the concentrations of covalent binding sites were obtained, which were relatively high in liver and kidney. Time periods for the hydrolysis of the isomer from a concentration of 40 ng/mL to 20 ng/mL were evaluated from the second reaction phase. It is concluded that the spontaneous and enzyme-catalyzed hydrolytic activities found for degradation of C(+)P(-)-soman in organs participating in central elimination are sufficiently high to account for the terminal half-life times of the isomer found in our toxicokinetic studies for the blood concentration after intoxication with 2-6 LD50 C(+/-)P(+/-)-soman. The hydrolytic activities are lower in the target organs for toxic action of soman, e.g., diaphragm and brain, especially for guinea pigs and marmosets. PMID- 8240391 TI - Chloroquine resistance in Plasmodium falciparum is not reversed by BIBW-22, a compound reversing the multidrug resistance phenotype in mammalian cancer cells. AB - The pteridine derivative BIBW-22 (4-[N-(2-hydroxy-2-methyl-propyl)-ethanolamino] 2,7-bis(cis-2,6-di methyl-morpholino)-6-phenylpteridine), which had been developed for the treatment of multidrug-resistant cancer and binds to P glycoprotein, was tested against chloroquine resistant Plasmodium falciparum strains in culture. Based on the result that BIBW-22 enhanced rather than lowered chloroquine resistance in vitro, it is concluded that chloroquine resistance in malaria parasites may not be mechanistically linked to the multidrug-resistant phenotype of chloroquine resistant P. falciparum. PMID- 8240392 TI - Inhibition of Ah (dioxin) receptor transformation by 9-hydroxy ellipticine. Involvement of protein kinase C? AB - 9-Hydroxy ellipticine (9-OHE), a metabolite of the anti-neoplastic agent ellipticine, is known to bind the aryl hydrocarbon (Ah) receptor in rat lung cytosol and to inhibit aryl hydrocarbon hydroxylase activity (AHH) in rat hepatic microsomes. In this study, the effects of 9-OHE on the transformation of the rat hepatic cytosolic Ah receptor to a form that binds the xenobiotic responsive enhancer element-3 (XRE-3) of the cytochrome P4501A1 gene was investigated. Sucrose density gradient analysis of [3H]-2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) binding in rat hepatic cytosol indicated that 9-OHE inhibited binding of the radiolabeled ligand to the Ah receptor with an IC50 of 90 microM. Gel retardation assays revealed that at low concentrations of 9-OHE the Ah receptor bound to XRE-3, as was the case with the TCDD-liganded receptor. However, in the presence of high concentrations of 9-OHE, the Ah receptor failed to transform to a form that could bind to XRE-3. In vitro studies indicated that incubation of rat hepatic cytosol with TCDD resulted in concentration-dependent increases in levels of protein kinase C (PKC) mediated phosphorylation as compared to vehicle treated extracts. Furthermore, 9-OHE concentrations that exhibited agonist activity with respect to Ah receptor transformation did not alter PKC phosphorylation in hepatic cytosol, whereas higher concentrations exhibited significant concentration-dependent decrease in PKC-mediated phosphorylation. These results demonstrate that the antagonistic effect of 9-OHE observed at high concentrations is due to inhibition of Ah receptor-XRE complex formation, a phenomenon that correlates with alterations in PKC activity. PMID- 8240393 TI - Protein phosphatase 2A and its [3H]cantharidin/[3H]endothall thioanhydride binding site. Inhibitor specificity of cantharidin and ATP analogues. AB - The target site for cantharidin (CA) and its analogues was isolated recently from mouse liver and identified as protein phosphatase (PP2A) in the heterodimeric form known as PP2A2. The most toxic CA analogue, endothall thioanhydride (ETA) (mouse i.p. LD50 0.3 mg/kg), appears to have the same binding site in mouse liver and brain based on studies comparing [3H]ETA and [3H]CA. ATP and its nonhydrolyzable analogues and pyrophosphate and related compounds including phosphonoformic acid inhibited both [3H]CA and [3H]ETA binding with IC50 values ranging from 2 to 81 microM. As with CA itself, the most potent inhibitors have two negatively charged groups in close proximity to each other. Inhibition of [3H]CA binding by 5,5'-dithiobis(2-nitrobenzoic acid) and stimulation by N ethylmaleimide indicated the involvement of a thiol site in the CA-binding domain. CA and three analogues (cantharidic acid, palasonin and endothall) inhibited PP2A and protein phosphatase 1 (PP1) but not PP2B or PP2C. The catalytic subunit of PP2A was 5- to 12-fold more sensitive to these CA analogues than the catalytic subunit of PP1. CA and the herbicide endothall also inhibited spinach leaf PP1 and PP2A and, at 50 microM, decreased the PP2A-mediated light induced activation of nitrate reductase in intact spinach leaves by 62 and 56%, respectively. This is consistent with PP2A as their site of action in plants, and indicates the potential use of CA analogues as pharmacological probes to investigate cellular processes that are regulated by reversible protein phosphorylation in vivo. PMID- 8240394 TI - Partial characterization of an approximately 20 K M(r) retinal protein whose phosphorylation is inhibited by taurine. AB - It has been demonstrated previously that taurine (2-aminoethanesulfonic acid) is an inhibitor of protein phosphorylation in a mitochondrial fraction of the rat retina. It appears that taurine is most effective in inhibiting the phosphorylation of an approximately 20 K apparent molecular weight (M(r)) protein found in the retinal tissue. This study further characterizes the location of the approximately 20 K phosphoprotein by phase separation using Triton X-114 and also characterizes the nature of the phosphate bond by various solvent extractions and by exposure to acid and base conditions. Triton X-114 experiments indicated that the approximately 20K phosphoprotein is located in the aqueous phase and, consequently, is probably not an integral protein of the mitochondrial membranes. Treatment of the phosphoprotein with solvents, acid, and/or base determined the phosphate linkage to be through a phosphoester bond rather than an acylphosphate bond. The approximately 20 K M(r) phosphoprotein was also isolated from one dimensional polyacrylamide gels and subsequently digested with trypsin and hydrolyzed with 1 M HCl to break all peptide bonds. Analysis of the phosphoamino acids by two-dimensional high voltage electrophoresis on cellulose plates revealed that it is both the serine and threonine residues that are phosphorylated. However, phosphorylation of the serine residue(s) is predominant. PMID- 8240395 TI - Elevated concentrations of dopamine sulfate in plasma of cocaine abusers. AB - This study investigated the effect of cocaine abuse on peripheral catecholamines. Specifically, we measured the concentration of free dopamine, dopamine sulfate, free norepinephrine, norepinephrine sulfate, free epinephrine and epinephrine sulfate in plasma samples obtained from the blood of a group of patients with cocaine addiction (N = 15). The concentrations of free and sulfoconjugated catecholamines in plasma were measured by a radioenzymatic technique. The results of this study revealed significant (P < 0.0001) elevation in plasma dopamine sulfate (8926 +/- 1204 pg/mL) of cocaine addicts upon admission to an in-patient treatment facility when compared with the level of this dopamine metabolite in plasma of control subjects (2356 +/- 121 pg/mL). Furthermore, there was a significant (P < 0.0001) relationship between elevation in plasma dopamine sulfate levels and severity of cocaine use among these patients, and in the majority of cases the plasma levels of dopamine sulfate declined appreciably in time with abstinence from cocaine. In contrast, no appreciable difference was observed in the concentrations of either free or sulfate-conjugated norepinephrine and epinephrine in plasma of cocaine addicts as compared with controls. Differences in plasma dopamine sulfate among these patients versus controls may be interpreted as a reflection of activation of extracellular dopamine metabolism associated with chronic cocaine exposure in humans. PMID- 8240396 TI - Carbonoyloxy analogs of the anti-metastatic drug swainsonine. Activation in tumor cells by esterases. AB - Swainsonine (SW), a plant alkaloid and inhibitor of alpha-mannosidases, has been shown to inhibit N-linked oligosaccharide processing and to block tumor cell metastasis in mice. In this study, a series of SW analogs were chemically synthesized and compared for inhibition of complex-type N-linked oligosaccharide processing in cultured MDAY-D2 tumor cells, for inhibition of alpha-mannosidases in vitro, and for stimulation of bone marrow proliferation in vivo. Carbonoyloxy substitutions at the 2 and 8 carbons of SW reduced inhibitor activity by 2-3 orders of magnitude for Jack Bean and MDAY-D2 tumor cell lysosomal alpha mannosidases in vitro. However, 2-p-nitrobenzoyloxy-, 2-octanoyloxy- and 2 butanoyloxy-derivatives of SW retained full activity as inhibitors of Golgi oligosaccharide processing in viable MDAY-D2 tumor cells. Inhibition of oligosaccharide processing was reduced by the esterase inhibitor diethyl p nitrophenyl phosphate, suggesting that although 2-p-nitrobenzoyloxy-SW, 2 octanoyloxy-SW and 2-butanoyloxy-SW are relatively poor inhibitors of alpha mannosidases in vitro, the compounds enter cells at a rate comparable to that of SW, and are converted to SW by cellular esterases. The more lipophilic esters, 2 benzoyloxy-SW, 2-toluoyloxy-SW, 8-palmitoyloxy-SW and 8-myristinoyloxy-SW, showed IC50 values at least 10 times higher for inhibition of Golgi oligosaccharide processing, probably due to less efficient entry of the compounds into tumor cells. The anti-metastatic activities of SW and two analogs were tested and shown to correlate with the IC50 values for inhibition of Golgi oligosaccharide processing in cultured tumor cells. In vivo, SW and the analogs were administered intraperitoneally to mice and found to have comparable activities as stimulators of bone marrow cell proliferation. Carbonoyloxy substitutions at the 2- or 8 position of SW with other chemical groups may lead to new drugs with improved pharmacokinetics and anti-cancer activity. PMID- 8240397 TI - A simple assay for ecto-5'-nucleotidase using intact pulmonary artery endothelial cells. Effect of endotoxin-induced cell injury. AB - Adenosine may be protective in acute vascular injury by inhibiting platelet aggregation and neutrophil oxidant release. In contrast, adenine nucleotides, which may be released with acute vascular injury, stimulate platelet aggregation and neutrophil oxidant release. Ectonucleotidases, membrane enzymes that catabolize extracellular nucleotides, are the primary mechanism for degrading circulating nucleotides to adenosine. Ecto-5'-nucleotidase converts extracellular AMP to adenosine. We hypothesized that endothelial cell injury alters ecto-5' nucleotidase activity. Using a novel assay first reported by Jamal et al. (Biochem J 250: 369-373, 1988) with rat adipocytes, we studied the properties of ecto-5'-nucleotidase in intact monolayers of cultured bovine pulmonary artery endothelial cells (BPAEC) and examined the effect of endotoxin on enzyme activity. The assay uses a fluorescent analog of AMP, 1,N6-etheno-AMP (E-AMP), as the substrate for ecto-5'-nucleotidase, and measures ethenoadenosine (E-Ado) formation. Etheno-AMP in Hepes buffer, pH 7.4, at 22 degrees, was added to confluent monolayers of BPAEC; samples of supernatant were collected after various intervals, and E-AMP and E-Ado were quantitated by HPLC. Using these methods we found a Km of 15 +/- 6 microM, a pH optimum of 7.48, minimal effect of MgCl2 or CaCl2 at physiologic pH, and inhibition by alpha,beta-methylene ADP, a known 5'-nucleotidase inhibitor. We established that the monolayer assay was indeed measuring cell surface associated 5'-nucleotidase. To determine the effect of endotoxin, we incubated confluent monolayers with endotoxin in Minimal Essential Medium plus 10% fetal bovine serum for 24 hr, washed them, and assessed the conversion of E-AMP to E-Ado by the endotoxin-injured cells. Endotoxin stimulated endothelial ecto-5'-nucleotidase activity. This increase in 5' nucleotidase activity in response to endotoxin injury may represent an important clearance mechanism for circulating adenine nucleotides and may be protective in acute vascular injury by increasing adenosine production. PMID- 8240398 TI - Effect of the lipid-lowering drug lifibrol on lipid metabolism in rat macrophages and in atherosclerotic arteries from swine and WHHL rabbits, in vitro. Implications in atherogenesis. AB - The effects of lifibrol on lipid metabolism in rat macrophages and swine and rabbit aortae were investigated. Resident peritoneal macrophages isolated from rats pretreated with lifibrol (50 mg/kg/7 days) showed a decreased capacity to synthesize cholesteryl esters from labeled precursors ([1-14C]oleate and [4 14C]cholesterol). Macrophages isolated similarly from non-treated rats demonstrated the ability to take up [14C]lifibrol, in vitro. Modification of lipid metabolism in atherosclerotic aortae from swine and Watanabe heritable hyperlipidemic (WHHL) rabbits was also observed when the tissues were incubated in vitro in the presence of exogenous lifibrol. Concentrations of lifibrol of up to 50 micrograms/mL in the incubations selectively reduced the formation of cholesteryl esters from [1-14C]acetate by 60-75%, whereas higher concentrations (100 micrograms/mL) resulted in a generalized inhibition of lipid biosynthesis of about 50% and of cholesteryl ester formation by up to 90%. The ability of lifibrol to directly affect these targets (i.e. macrophages and arterial tissue) has implications that extend beyond its confirmed plasma cholesterol-lowering activity since early stages of the atherogenic process involve an overall increase in arterial lipid synthesis and cholesteryl ester accumulation by monocyte-macrophages that infiltrate the vessel wall from blood. PMID- 8240399 TI - Biochemical characterization of a membrane-bound enzyme responsible for generating nitric oxide from nitroglycerin in vascular smooth muscle cells. AB - A membrane-bound enzyme responsible for generating nitric oxide (NO) from nitroglycerin (NTG) in vascular smooth muscle cells has been characterized. The enzyme could be solubilized from vascular microsomes by several detergents, the most effective of which was 3-[(3-cholamidopropyl)-dimethylamino]-1 propanesulfonate (CHAPS). A partially purified enzyme preparation was obtained with CHAPS-solubilized vascular microsomes that were processed sequentially through an ion exchange column and a gel filtration column. The activity of this partially purified enzyme showed a dependence on substrate concentration, protein concentration and the duration of incubation. Enzyme activity was enhanced 2.7- to 4.2-fold by several thiols such as cysteine, N-acetylcysteine, reduced glutathione, and dithiothreitol. On the other hand, N-ethylmaleimide, iodoacetic acid, p-chloromercuric benzoic acid and 1-chloro-2,4-dinitrobenzene, reagents known to bind with the free sulfhydryl groups, inactivated the NO-generating activity from NTG. The enzyme activity could be reversibly bound to an organomercurial column. These results suggested the presence of a free thiol group in the enzyme and that this thiol group was required for enzyme activity. The partially purified enzyme was active in the presence of 0.1% sodium dodecyl sulfate (SDS). The enzyme was purified to near homogeneity using several sequential chromatographic steps including DEAE-Sephacel, Biogel A 1.5 m, hydroxylapatite and organomercurial columns, resulting in an increase in enzyme activity of about 94-fold. The subunit of this enzyme, as identified on an SDS treated electrophoresis gel, had an apparent molecular size of 58 kDa. PMID- 8240400 TI - Inhibition by the protein kinase inhibitor HA1077 of the activation of NADPH oxidase in human neutrophils. AB - The effect of an inhibitor of protein kinase, HA1077 [1-(5-isoquinolinesulfonyl) homopiperazine HCl], and its hydroxylated metabolite, HA1100, on the activation of NADPH oxidase in human neutrophils were studied. Cells were preincubated with each drug for 10 min and then activated by treatment with phorbol myristate acetate (PMA) or formylmethionyl leucyl phenylalanine (FMLP). After activation, the rate of superoxide dismutase-inhibitable reduction of cytochrome c was estimated. HA1077 and HA1100 inhibited the PMA-induced production of O2- by neutrophil NADPH oxidase in a concentration-dependent manner (IC50 = 15 and 24 microM, respectively). The sensitivity of the FMLP-induced production of O2- to these drugs was similar. The production of O2- in 1,25-dihydroxyvitamin D3 treated HL-60 cells, which differentiated to macrophage-like cells, was also inhibited by the drugs. The extent of inhibition by HA1077 was almost the same as that by a calmodulin inhibitor (W-7) and by inhibitors of protein kinase (H-7 and H-8). In a cell-free lysate of neutrophils, the NADPH-dependent production of O2- can be induced by sodium dodecyl sulfate (SDS). HA1077 at 100 microM had only a weak inhibitory effect on the cell-free, SDS-induced production of O2-, an indication that HA1077 inhibits the activation of NADPH oxidase, not the actual activity. The effects of H-7 and H-8 were similar to that of HA1077, whereas W-7 inhibited the production of O2- by the cell-free extract of HL-60 cells. This action of HA1077 could explain, in part, its ability to protect neuronal cells from death after ischemia. PMID- 8240401 TI - Mechanisms of protection from menadione toxicity by 5,10-dihydroindeno[1,2, b]indole in a sensitive and resistant mouse hepatocyte line. AB - Established cell lines derived from newborn livers of c14CoS/c14CoS and cch/cch mice have been shown to be genetically resistant (14CoS/14CoS cells) or susceptible (ch/ch cells) to menadione toxicity. These differences are due in part to relatively higher levels of reduced glutathione (GSH) and NAD(P)H:menadione oxidoreductase (NMO1) activity in the 14CoS/14CoS cells. The indolic membrane-stabilizing antioxidant 5,10-dihydroindeno[1,2-b]indole (DHII) was shown previously to protect against various hepatotoxicants in vivo and in primary rat hepatocytes. This report describes how the 14CoS/14CoS and ch/ch cell lines provide a valuable experimental system to distinguish the mechanism of chemoprotection by DHII from menadione toxicity. The addition of 25 microM DHII produced a time-dependent decrease in menadione-mediated cell death in 14CoS/14CoS cells, with little effect on ch/ch cell viability. The maximum protective effect occurred at 24 hr, although the concentration of DHII remained constant for 48 hr. The protective effect of DHII correlated with enhanced glutathione levels (234% increase at 24hr), as well as induction of four enzymes involved in the detoxification and excretion of menadione: NAD(P)H:menadione oxidoreductase (NMO1, quinone reductase), glutathione reductase, glutathione transferase (GST1A1), and UDP glucuronosyltransferase (UGT1*06), with 24-hr maximum induction of 707, 201, 171 and 198%, respectively. Other biotransformation enzymes not directly involved in menadione metabolism (glutathione peroxidase, cytochromes P4501A1 and P4501A2, copper-, zinc-dependent superoxide dismutase, and NADPH cytochrome c oxidoreductase) were not induced by DHII. Menadione-stimulated superoxide production was inhibited 50% by DHII only in 14CoS/14CoS cells, and the inhibition required 24-hr preincubation. Pretreatment with DHII also protected both cell types against the menadione mediated depletion of GSH, and the increase in percent (oxidized glutathione GSSG), an indicator of oxidative stress. These results suggest that DHII does not protect against menadione toxicity by virtue of its antioxidant or membrane stabilizing properties. Rather, it acts by inducing a protective enzyme profile that migates redox cycling and facilitates excretion of menadione. PMID- 8240402 TI - Inhibition of lipogenesis in rat brown adipose tissue by clofibrate. AB - The effect of clofibrate (Atromid S, ethyl-2-(4-chlorophenoxy)-2 methylpropionate) administration for 7 days to rats on lipogenesis and on some lipogenic enzyme activities in brown adipose tissue (BAT), liver and white adipose tissue (WAT) was examined. As compared to control rats the rate of lipogenesis in BAT in the clofibrate-treated animals was significantly decreased. The rate of liver lipogenesis increased slightly, whereas lipogenesis in the WAT was not affected by clofibrate. In BAT, the drug treatment resulted in depression of fatty acid synthase, ATP-citrate lyase, malic enzyme, glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities. The activity of liver fatty acid synthase did not change, ATP-citrate lyase activity slightly decreased, whereas the activity of malic enzyme significantly increased in this organ after clofibrate feeding. The ATP-citrate lyase activity in WAT decreased, while fatty acid synthase and other lipogenic enzymes were not changed after clofibrate feeding. Clofibrate treatment did not influence the activity of NADP linked isocitrate dehydrogenase and malate dehydrogenase (enzymes not linked directly to lipogenesis), either in BAT, liver or WAT. The data presented suggest that the hypolipidaemic effect of clofibrate in the rat may be due (possibly among other mechanisms) to reduction of the rate of fatty acid synthesis in BAT but not in the liver and WAT. PMID- 8240403 TI - Excretion balance and urinary metabolites of the S-enantiomer of indobufen in rats and mice. AB - The excretion balance and urinary metabolites of the S-enantiomer of indobufen, ((S)2-[p-(1-oxo-2-isoindolinyl)-phenyl]butyric acid), a platelet aggregation inhibitor, were studied in rats and mice after oral administration. The urinary metabolic profile exhibited a marked species difference. The major metabolic pathway in the mouse was acyl glucuronidation followed by renal excretion, whereas in rat urine 5-hydroxylation and subsequent sulphation at the introduced hydroxyl group accounted for almost all recovered radioactivity. Indobufen glucuronide was the major biliary metabolite in the rat, while very little indobufen glucuronide was present in the urine of intact or bile duct-cannulated rats. A marked dose-effect on the elimination and metabolism of S-indobufen was demonstrated in the rat. The recovery (% dose) of 5-hydroxyindobufen and its sulphate after the lower dose of the enantiomer (10 mg/kg) was some 2.8-fold higher compared with the higher dose of 20 mg/kg. PMID- 8240404 TI - Effects of sodium fluoride and cobalt chloride on the enantioselectivity of microsomal and cytosolic esterases in rat intestinal mucosa. AB - The effects of sodium fluoride (NaF) and cobalt chloride (CoCl2) on the enantioselective hydrolysis of racemic oxazepam 3-acetate (rac-OXA) by microsomal and cytosolic esterases in rat intestinal mucosa were studied. Microsomal and cytosolic esterases hydrolyzed S-OXA and R-OXA in approximately 1:19 and 4:1 ratios, respectively. The hydrolysis of R-OXA by microsomal esterases was inhibited by NaF with an IC50 of 13.4 +/- 1.5 mM. Hydrolyses of both S-OXA and R OXA by cytosolic esterases were inhibited by NaF with a similar IC50 value (approximately 3 mM). The hydrolysis of S-OXA by cytosolic esterases was inhibited by CoCl2 (IC50 = approximately 5 mM), whereas the hydrolysis of R-OXA by cytosolic esterases was stimulated by approximately 10% in the presence of 1 mM CoCl2. In comparison, the hydrolysis of R-OXA by microsomal esterases was stimulated by approximately 55% in the presence of 1 mM CoCl2. These results not only revealed the effects of NaF and CoCl2 on the catalytic activities of enantioselective cytosolic and microsomal esterases, but also indicated that microsomal and cytosolic esterases that selectively hydrolyzed R-OXA were distinctly different protein entities. PMID- 8240405 TI - Gemfibrozil modifies acyl composition of liver microsomal phospholipids from guinea-pigs without promoting peroxisomal proliferation. AB - Treatment with gemfibrozil modifies acyl composition of hepatic microsomal phosphatidylcholine and phosphatidylethanolamine in guinea-pigs. Palmitic (16:0) and palmitoleic (16:1) fatty acids are increased, and stearic (18:0) and oleic (18:1) are decreased; further, while linoleic acid [18:2 (n-6)] is increased by gemfibrozil treatment, the other constituents of the n-6 fatty acids family, including arachidonic acid [20:4 (n-6)], are decreased. As gemfibrozil is a potent inhibitor of fatty acid elongation in vitro (Sanchez et al., FEBS Lett 300: 89-92, 1992), the inhibition of this enzyme system by gemfibrozil treatment could be responsible for the observed results in vivo. These changes in fatty acid composition are accompanied by a decrease in serum lipids and, more important, are independent of peroxisomal proliferation. PMID- 8240406 TI - Does the fluidity of the lipid environment modulate membrane-bound acetylcholinesterase? Effects of temperature, membrane composition and amphiphiles. PMID- 8240407 TI - An investigation of the formation of cytotoxic, genotoxic, protein-reactive and stable metabolites from naphthalene by human liver microsomes. AB - Chemically reactive epoxide metabolites have been implicated in various forms of drug and chemical toxicity. Naphthalene, which is metabolized to a 1,2-epoxide, has been used as a model compound in this study in order to investigate the effects of perturbation of detoxication mechanisms on the in vitro toxicity of epoxides in the presence of human liver microsomes. Naphthalene (100 microM) was metabolized to cytotoxic, protein-reactive and stable, but not genotoxic, metabolites by human liver microsomes. The metabolism-dependent cytotoxicity and covalent binding to protein of naphthalene were significantly higher in the presence of phenobarbitone-induced mouse liver microsomes than with human liver microsomes. The ratio of trans-1,2-dihydrodiol to 1-naphthol was 8.6 and 0.4 with the human and the induced mouse microsomes, respectively. The metabolism dependent toxicity of naphthalene toward human peripheral mononuclear leucocytes was not affected by the glutathione transferase mu status of the co-incubated cells. Trichloropropene oxide (TCPO; 30 microM), an epoxide hydrolase inhibitor, increased the human liver microsomal-dependent cytotoxicity (19.6 +/- 0.9% vs 28.7 +/- 1.0%; P = 0.02) and covalent binding to protein (1.4 +/- 0.3% vs 2.8 +/- 0.2%; P = 0.03) of naphthalene (100 microM), and reversed the 1,2-dihydrodiol to 1-naphthol ratio from 6.6 (without TCPO) to 2.6, 0.6 and 0.1 at TCPO concentrations of 30, 100 and 500 microM, respectively. Increasing the human liver microsomal protein concentration reduced the cytotoxicity of naphthalene, while increasing its covalent binding to protein and the formation of the 1,2 dihydrodiol metabolite. Co-incubation with glutathione (5 mM) reduced the cytotoxicity and covalent binding to protein of naphthalene by 68 and 64%, respectively. Covalent binding to protein was also inhibited by gestodene, while stable metabolite formation was reduced by gestodene (250 microM) and enoxacin (250 microM). The study demonstrates that human liver cytochrome P450 enzymes metabolize naphthalene to a cytotoxic and protein-reactive, but not genotoxic, metabolite which is probably an epoxide. This is rapidly detoxified by microsomal epoxide hydrolase, the efficiency of which can be readily determined by measurement of the ratio of the stable metabolites, naphthalene 1,2-dihydrodiol and 1-naphthol. PMID- 8240408 TI - Effects of chronic benfluorex treatment on the activities of key enzymes of hepatic carbohydrate metabolism in old Sprague-Dawley rats. AB - Chronic effects of benfluorex on some parameters of carbohydrate metabolism have been studied in 24-month-old Sprague-Dawley rats. Treatment once a day for 14 days with 25 mg benfluorex per kg body weight lowered body weight, decreased circulating insulin and resulted in an increase in hepatic glycogen. Measurement of the activities of several important regulatory enzymes of hepatic carbohydrate metabolism showed a significant decrease in the activities of phosphoenolpyruvate carboxykinase and glycogen phosphorylase. The activity of glucose-6-phosphatase, on the other hand, was slightly increased. Taken collectively, our data offer an explanation for the observed inhibition of hepatic glucose production by chronic benfluorex treatment in cases of hyperinsulinemia. PMID- 8240409 TI - Inhibition of the human leukocyte endopeptidases elastase and cathepsin G and of porcine pancreatic elastase by N-oleoyl derivatives of heparin. AB - N-oleoyl-heparin derivatives differing in their oleic acid and sulfate contents were synthesized and studied for their abilities to inhibit human leukocyte elastase (HLE), human leukocyte cathepsin G (CatG) and porcine pancreatic elastase (PPE) at pH 8.0, ionic strength 0.05 M and 37 degrees. Heparin (Hep) as well as N-oleoyl-heparins behaved as tight-binding, hyperbolic noncompetitive inhibitors of HLE (KiHep = 75 pM) and CatG (KiHep < 25 pM). The main driving force for the interaction between enzymes and glycosaminoglycans was electrostatic in nature. Under the condition [enzyme] >> Ki, the stoichiometries of the interaction with Hep were 1:2 (Hep:HLE) and 1:4 (Hep:CatG). Coupling one oleic acid residue to three disaccharide units of partially N-desulfated Hep, Ol1:3Hep, lowered HLE inhibition (Ki = 0.3 nM) and the stoichiometry of binding was reduced to 1:1. Re-N-sulfation of a similar derivative, Ol1:5Hep(SO4), containing one fatty acid residue for five disaccharide units, led to a substance with similar HLE inhibitory characteristics as Hep (Ki = 92 pM) and stoichiometry 1:2. Ol1:5Hep(SO4) was also a more efficient inhibitor of CatG (Ki < 33 pM) than Ol1:3Hep (Ki = 9.5 nM). The residual activities of N-oleoyl-Hep complexes with CatG were much lower than the corresponding activities in the presence of Hep. While oleate and Hep could not inhibit PPE, N-oleoyl-Hep, independently of fatty acid substitution and sulfate content, could inhibit this enzyme with Ki congruent to 60 nM and low residual activity. The efficient endopeptidase inhibitory characteristics of N-oleoyl-Hep derivatives, together with their non anticoagulant properties and their capacity to interact with elastin, may be therapeutically useful in connective tissue degenerative diseases. PMID- 8240410 TI - Stimulation of mucus glycoprotein biosynthesis in rat gastric mucosa by gastrin. AB - We examined the effects of the gastrin family of peptides on gastric mucus glycoprotein (mucin) biosynthesis in rat gastric mucosa using an organ culture technique. Radiolabeled mucin was obtained from the tissue and culture medium of the corpus and antrum of rat stomach incubated for 5 hr with [3H]glucosamine (GlcN), [14C]threonine (Thr), and [35S]sulfate in vitro. With the addition of 10( 8) and 10(-7) M tetragastrin to the culture medium, [3H]GlcN labeled mucin in the corpus tissue increased to 120-135% that of the control (P < 0.01). The biosynthetic responses to cholecystokinin (CCK)-8 and the 17-peptide gastrin were essentially the same as that to tetragastrin. Tetragastrin 10(-8) M also increased the incorporation of [35S]sulfate into the corpus mucin but failed to change [14C]Thr incorporation. In the antrum, biosynthetic activity showed no significant change with 10(-9) approximately 10(-5) M tetragastrin. Ranitidine, diphenhydramine and omeprazole at 10(-5) M did not suppress the tetragastrin induced increase in [3H]GlcN incorporation into mucin, but L-365,260 at a concentration of 10(-6) M completely blocked this effect. These results suggest that gastrin stimulates mucin production via CCK-B/gastrin receptors in the oxyntic region of rat gastric mucosa. PMID- 8240411 TI - Characterization of adenylyl cyclase in goldfish brain. AB - The rate of production of cAMP by the adenylyl cyclase enzyme from goldfish brain was linear with time and with protein concentration. In agreement with mammalian adenylyl cyclase systems the enzyme is divalent cation dependent, being activated in the presence of either Mg2+ or Mn2+. Forskolin also stimulated the rate of reaction in a dose-dependent manner with a half-maximal effect of 1 microM. The activated enzyme was inhibited by high concentrations of Ca2+ but was independent of Na+ concentration. The presence of guanine nucleotide binding proteins (G proteins) was demonstrated by the fact that both NaF and guanosine 5'-[beta gamma imido]triphosphate (p[NH]ppG) stimulated the basal rate. In addition, the p[NH]ppG dose-response curve of the forskolin-stimulated enzyme was biphasic, similar to that observed for other systems. At low concentrations of p[NH]ppG a small inhibition was observed while higher concentrations produced a stimulation. These data suggest that the goldfish brain adenylyl cyclase enzyme complex includes both stimulatory and inhibitory G-proteins in addition to the catalytic unit. A series of known and putative goldfish neurotransmitter substances failed to either stimulate or inhibit the adenylyl cyclase activity. The endogenous neurotransmitters which interact with this second messenger system remain to be determined. PMID- 8240412 TI - Dexamethasone treatment fails to reduce oxygen-induced lung injury in the preterm guinea pig. Effects on pulmonary inflammation and antioxidant status. AB - Dexamethasone (10 mg/kg/day) or vehicle was administered in a randomized, controlled fashion to 3-day preterm guinea pigs exposed to either 21% oxygen or 95% oxygen for 72 hr and maintained in room air for a further 96 hr. Treatment with dexamethasone had no effect on survival of preterm pups maintained in either 21% or 95% O2. Dexamethasone treatment reduced the growth rate of pups, the effect occurring earlier (0-3 days) in 21% O2-treated pups than in 95% O2-treated pups (5-7 days). Exposure to 95% O2 reduced the survival rate of preterm animals (73% vs 100%, P < 0.05). Surviving pups developed acute lung injury, characterized by the accumulation of a protein-rich exudate in the alveoli and an infiltration of inflammatory cells, particularly neutrophils into the lung. Dexamethasone treatment attenuated the pulmonary inflammatory cell infiltration, in particular neutrophils, both during oxygen exposure (16.4 x 10(4) vs 9.4 x 10(4)/mL; P < 0.05) and following return to ambient conditions (28.0 x 10(4) vs 5.1 x 10(4)/mL; P < 0.05). Elastase activity in bronchoalveolar lavage fluid, which was primarily of neutrophil origin, was unchanged by dexamethasone treatment. Dexamethasone-treated pups had increased pulmonary antioxidant enzyme activities (Cu/Zn-superoxide dismutase; Mn-superoxide dismutase, catalase and glutathione peroxidase) during recovery from oxidative injury. Although there was both a marked reduction in numbers of neutrophils in the lung and elevated pulmonary antioxidant enzyme activities in dexamethasone-treated pups, the degree of microvascular permeability, as determined by both the lung wet weight/dry weight ratio and the presence of plasma proteins in the lavage fluid, was unchanged. Combined, these results imply that dexamethasone, although capable of blunting the influx of neutrophils to the hyperoxia-exposed lung and inducing antioxidant defences in the immature lung, cannot modify the progression of acute oxygen-induced injury of the immature lung. PMID- 8240413 TI - Further evidence for the interaction of the antimalarial drug amodiaquine with ferriprotoporphyrin IX. AB - Evidence for complex formation of the antimalarial drug amodiaquine (AD) with ferriprotoporphyrin IX (FP) in aqueous medium is presented, in addition to previous preliminary data. A mole ratio of one between the complex components is determined for the insoluble complex at pH 6.7-6.8. Mossbauer data obtained at pH 7-8 and at higher concentrations in the millimolar range confirm the interactions existing between the complex components. These data are considered to aid in removing previous objections to a mechanism of antimalarial action involving complexes of FP with AD and related drugs. PMID- 8240414 TI - A highly sensitive tool for the assay of cytochrome P450 enzyme activity in rat, dog and man. Direct fluorescence monitoring of the deethylation of 7-ethoxy-4 trifluoromethylcoumarin. AB - The O-deethylation of 7-ethoxy-4-trifluoromethylcoumarin (EFC) by liver microsomes has been assessed as a method for monitoring the activity of cytochrome P450. The principle advantage of this substrate is the formation of a fluorescent product 7-hydroxy-4-trifluoromethylcoumarin (HFC) which can be assayed directly in the reaction medium. For rat microsomes the deethylated product was confirmed as the main metabolite, the reaction rate was linear with respect to both time and microsomal protein concentration and was independent of small changes in the added co-factors. A linear formation rate for the deethylated metabolite was also confirmed with dog and human microsomes. The intra-assay precision for rat, dog and human microsomes was 3, 5 and 4%, respectively. Hanes transformations of the dog and human data showed two phases, in contrast to a linear decline seen for the rat. Hybrid parameters for Vmax and Km, calculated from the apparently linear portions of these curves, gave interday SD for the Vmax of rat, dog and man of 2, 14 and 4%, respectively, and approximately 15% for the Km in all species. The Vmax in rat, dog and human microsomes was 1.4 +/- 0.2, 4.3 +/- 1.5 and 0.9 +/- 0.5 nmol HFC/min/nmol P450, respectively. The Km was 11.0 +/- 3.1, 67 +/- 19 and 6.8 +/- 2.5 microM, respectively. Direct evidence that at least two isoenzymes (cytochrome P450 1A2 and 2E1) metabolize EFC was obtained by experiments with competitive, suicide and immuno-inhibitors. Compared with ethoxycoumarin, the involvement of P450 2E1 in O deethylation seemed similar in the rat. In conclusion, EFC provides a straightforward and reproducible assay for microsomal enzyme activity, requiring at most 25 pmol/mL of cytochrome P450. PMID- 8240415 TI - Increase in insulin binding and inhibition of the decrease in the phospholipid content of human term placental homogenates in culture by the sulfonylurea glipizide. AB - Term placental explants were cultivated for 48 hr without (control) and with various concentrations of glipizide. Maximum binding of [125I]-insulin in the control samples was decreased after 12 and 24 hr returning to initial values after 48 hr. In the presence of glipizide the binding was generally higher, reaching 180% (557 and 1000 nmol/L) of the corresponding control value (P < 0.01) after 48 hr owing to the presence of nearly 3-fold more (P < 0.05) receptors than in the untreated controls. Tissue cholesterol content was almost unaffected whereas both the phospholipid content and the corresponding phospholipid-to cholesterol ratios were markedly, and in a time-dependent manner, increased by glipizide as compared to the controls. This was due to decreasing cholesterol and phospholipid concentrations in the controls during the time of culture as compared to initial values, and also to unchanged levels in glipizide-treated cultures. We conclude that glipizide affects placental insulin receptors and the phospholipid content of the tissue. PMID- 8240416 TI - Importance of the paracellular pathway for the transport of a new bisphosphonate using the human CACO-2 monolayers model. AB - The transport of a new bisphosphonate, Tiludronate, was investigated on the human adenocarcinoma cell line, CACO-2. Experiments were performed 7-16 days after cells achieved confluence, conditions under which they form well-differentiated monolayers joined by tight junctions. Tiludronate transport rate across CACO-2 monolayers was independent of the temperature (4 degrees versus 37 degrees), of the polarity of the cell membrane (apical-to-basolateral versus basolateral-to apical), and of the presence of metabolic poisons (sodium azide). Its transport was enhanced by either the presence of EGTA in the incubation buffer, i.e. when extracellular Ca2+ concentration was reduced, or by the pretreatment of monolayers with EGTA, i.e. when the intercellular spaces and the tight junctions were widened. Based on these different observations, we could suggest that Tiludronate mainly used the paracellular pathway to cross the intestinal epithelium. An increase in the Tiludronate permeability coefficient was also observed following treatment of cells with high Tiludronate concentrations, as a consequence of the direct effect of this compound on the extracellular Ca2+ ions. Hence, for high drug concentrations, i.e. 20 mM, we observed a decrease in free extracellular Ca2+ concentration, an increase in the transepithelial electrical resistance and an increase in the transport of [14C]polyethyleneglycol ([14C]PEG400), a probe for the paracellular pathway. The results indicate that Tiludronate is transported across CACO-2 monolayers by the paracellular route. Moreover, it can affect its own transport by its concentration-dependent effect on tight junction widening. PMID- 8240417 TI - Use of pharmacological agents to implicate a role for phosphoinositide hydrolysis products in malaria gamete formation. AB - The kinetics of phosphoinositol 4,5 bisphosphate hydrolysis products in activated Plasmodium falciparum gametocytes suggests a role for inositol trisphosphate [Ins(1,4,5)P3] and diacylglycerol (DAG) in the signal transduction pathway of malaria gametocytes. To investigate further this role, compounds that have an effect on the metabolism and biologic functions of these second messengers were tested in an in vitro system. Gentamycin, 2,3 diphosphoglycerate (2,3 DPG) and magnesium ion (Mg2+), inhibitors of Ins(1,4,5)P3 5' phosphatase, all stimulated gametocytes to exflagellate in suspended animation buffer, pH 7.4, at room temperature. In addition, methylxanthines, caffeine and theobromine, calcium ionophore (A-23187), and external calcium also stimulated exflagellation. In contrast, neomycin, an aminoglycoside that inhibits phospholipase C activity, and heparin, an antagonist of Ins(1,4,5)P3 binding to its receptor, inhibited microgamete formation. Quinine and chloroquine which can inhibit both phospholipase A and C activity also inhibited gametocyte exflagellation. The consistent manner in which these various compounds affect gametocyte activation further implicates phosphoinositol turnover in the signal transduction pathway of falciparum gametocytes. PMID- 8240418 TI - Characterization of receptor-mediated actions of T-kinin. AB - T-Kinin (Ile-Ser-bradykinin) is unique to the rat. This study characterizes the receptors involved in T-kinin activity on both the intact isolated rat uterus and membrane receptor preparations of the rat uterus. The results show that T-kinin acts through kinin B2 receptors in the rat uterus as demonstrated by B2 receptor antagonist inhibition. While the potency of T-kinin on rat uterus contraction was similar to that of bradykinin, binding studies showed that the affinity of T kinin to the receptor was 10-fold lower than that of bradykinin. On the other hand, the D isomer of T-kinin, D-Ile-Ser-bradykinin, had an affinity for the receptor greater than that of T-kinin and was more potent in causing contraction. Comparing this finding with our previously published report that D-Ile-Ser bradykinin is not active on the kinin receptor for vascular permeability indicates that the kinin receptors in the rat uterus are not the same as those previously reported in the smooth muscle of the vasculature, i.e. there exists subclasses of kinin B2 receptors. The data from binding studies on a variety of T kinin analogues show that the substitution of hydroxyproline (Hyp) for Pro5, together with the D-configuration at Ile1 and/or Ser2 may be useful for the development of selective T-kinin antagonists. Studies involving pretreatment of the tissue with indomethacin demonstrated that prostaglandin release was more of a component of T-kinin's activity on the rat uterus than that of bradykinin. PMID- 8240419 TI - Molecular mechanisms of chloroacetaldehyde-induced cytotoxicity in isolated rat hepatocytes. AB - 2-Chloroacetaldehyde (CAA) induced a loss in hepatocyte viability in a concentration- and time-dependent manner. Three phases before cytotoxicity ensued could be distinguished. Glutathione (GSH) was depleted immediately upon addition of CAA but only partial depletion occurred with subtoxic CAA concentrations. GSH depleted hepatocytes were much more susceptible to CAA toxicity, indicating that CAA was detoxified by GSH. The second phase of changes involved a steady decrease in protein thiol levels, mitochondrial respiration, transmembrane potential and ATP levels. The third phase involved lipid peroxidation which commenced at around 60 min with a CAA concentration that caused 50% cytotoxicity in 120 min. Addition of antioxidants (diphenylphenylenediamine, butylated hydroxyanisole) and iron chelators (desferoxamine) at 40 min prevented lipid peroxidation and delayed CAA induced cytotoxicity without restoring protein thiols, hepatocyte respiration or preventing further ATP depletion. Addition of dithiothreitol at 40 min, however, restored protein thiols and hepatocyte respiration, and prevented further ATP depletion and cytotoxicity. CAA-induced hepatocyte cytotoxicity therefore involved reversible thiol protein adduct formation, mitochondrial toxicity and lipid peroxidation. PMID- 8240420 TI - Desulfuration of 6-mercaptopurine. The basis for the paradoxical cytotoxicity of thiopurines in cultured human leukemic cells. AB - The thiopurines have a wide array of effects on purine metabolism, but the primary mechanism of cytotoxicity for both 6-mercaptopurine (6-MP) and 6 thioguanine (6-TG) appears to be incorporation of drug into DNA following conversion to the thioguanylate form. In murine leukemic cell lines exposed to a range of thiopurine concentrations in vitro, cell survival curves have displayed a phenomenon termed paradoxical cytotoxicity, defined as a decrease in cytotoxicity with increasing drug concentration. The paradoxical cytotoxicity of thiopurines has usually been attributed to concentration-dependent perturbations in the cell cycle. The present study assessed whether the paradoxical cytotoxicity of 6-MP occurred in cultured human leukemic cells, and investigated the biochemical and cell-cycle alterations occurring in these lines at thiopurine concentrations associated with the reverse of cytotoxicity. Paradoxical cytotoxicity was observed in the two human leukemic cell lines examined, but only when 6-MP concentrations exceeded 100 microM. The extent of incorporation of 6-MP metabolites into DNA as thiol-versus non-thiol-containing metabolites was analyzed by performing parallel experiments with 14C- and 35S-radiolabeled drug. With 5 microM 6-MP, approximately 50% of drug was incorporated into DNA as a thionucleotide; however, with increasing drug concentrations, the degree of thionucleotide incorporation remained unchanged or decreased, and the amount incorporated as the desulfurated metabolite (presumably adenylate or guanylate) increased. With 500 microM 6-MP, less than 10% of the drug was incorporated as the thionucleotide. Perturbations in cell cycle reflected the relative amounts of thiol- and non-thiol-containing nucleotide formed at various concentrations of 6 MP. These results suggest that thiopurines may be vulnerable to a unique mechanism of detoxification, in which a human cell can metabolize a cytotoxic drug to a comparatively potent "self-rescue" agent. PMID- 8240421 TI - 7-S-glutathionyl-tryptamine-4,5-dione: a possible aberrant metabolite of serotonin. AB - Tryptamine-4,5-dione (Compound 1) is an in vitro oxidation product of 5 hydroxytryptamine (5-HT). Recent evidence has suggested that aberrant oxidations of 5-HT occur in the central nervous system of individuals with Alzheimer's disease (AD). In the event that Compound 1 is formed as a result of oxidation of 5-HT within serotonergic nerve terminals or axons, it would be expected to be rapidly conjugated by intraneuronal glutathione (GSH) to give 7-S-glutathionyl tryptamine-4,5-dione (Compound 2). When injected into the brains of laboratory mice, Compound 2 was lethal (LD50 = 21 micrograms) and evoked hyperactivity for the first 30 min following drug administration. Particularly during this hyperactive phase Compound 2 caused a statistically significant decrease in whole brain levels of norepinephrine and 5-HT. Levels of dopamine were also decreased while whole brain concentrations of its metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid, were increased significantly. In the presence of GSH, NADPH and ascorbic acid, Compound 2 redox cycled in reactions that catalyzed the oxidation of these cellular reductants by molecular oxygen and formed H2O2 as a byproduct. Compound 2 also reacted with molar excesses of GSH to form more structurally complex glutathionyl conjugates. Several of these conjugates have been isolated and their structures determined using spectroscopic methods. It is conceivable that one or more of these conjugates might serve as analytical markers in a search for evidence in support of the hypothesis that aberrant oxidations of 5-HT occur in the Alzheimer brain. The redox cycling properties of Compound 2 and its facile reactions with cellular nucleophiles such as GSH may represent mechanisms that contribute to the toxicity of this drug. PMID- 8240422 TI - Angiotensin II and non-angiotensin II displaceable binding sites for [3H]losartan in the rat liver. AB - By virtue of the more than 1000-fold selectivity of losartan (DuP 753) for the AT1 angiotensin II (AII) receptor subtype compared with the AT2 subtype, [3H]losartan may be a useful radioligand for studies of the AT1 receptor subtype. Comparison of Bmax values in the liver obtained from saturation isotherms using [3H]losartan (Bmax = 194 pmol/g tissue) and [125I]sarcosine1,isoleucine8 angiotensin II (Bmax = 20 pmol/g tissue) indicated that the AII receptor concentration was approximately 10% that of the [3H]losartan binding sites. In addition, AII at concentrations as high as 10 microM displaced less than one third of specific [3H]losartan binding in the liver and less than 80% in the whole adrenal. The presence of non-AII displaceable [3H]losartan binding in the liver did not appear to result from metabolism of the radioligand since HPLC analysis of free and bound 3H revealed that greater than 90% of the 3H eluted at the same time as the parent [3H]losartan. This suggests that [3H]losartan binds with high affinity to a site(s) other than angiotensin II receptors in the rat liver. PMID- 8240423 TI - Thermodynamics of the interaction of berberine with DNA. AB - The interaction of berberine [7,8,13,13a-tetrahydro-9-10-dimethoxy-2,3-(methylene dioxy)-berberinium] with calf thymus DNA has been studied by spectrophotometry in buffers of various salt concentrations and temperatures. Binding parameters obtained are best fit by the neighbour exclusion model. The salt and temperature dependence of the binding constants are used to estimate thermodynamic parameters involved in the complex formation of berberine with DNA. The binding process is exothermic over the entire range and the values of enthalpy and entropy change are strongly dependent on the salt concentration. The negative enthalpy and positive entropy changes compensate one another to produce a relatively small Gibbs' free energy change. Possible molecular contribution to the enthalpy and entropy changes is discussed. PMID- 8240424 TI - Inhibition of in vitro mineralization in osteoblastic cells and in mouse tooth germ by phosphatidylinositol-specific phospholipase C. AB - We have examined the effect of phosphatidylinositol-specific phospholipase C (PIPLC) on the in vitro mineralization during cultivation of both osteoblast-like MC3T3-E1 cells and the lower molar tooth germ from mouse embryo. PIPLC not only caused the release of alkaline phosphatase (ALP) from cell membranes into the culture medium but also produced a disturbance of in vitro mineralization in MC3T3-E1 cells and in mouse embryonic tooth germ. These findings strongly suggest that ALP, functioning as an ectoenzyme, is involved in the physiological mineralization process of both bone and teeth. PMID- 8240425 TI - Effects of C-methylated carnitine analogs on rates of mitochondrial fatty acid oxidation. AB - Carnitine analogs containing one and/or two methyl substituents on the alpha-, beta-, or gamma-carbon were evaluated in isolated rat liver mitochondria for their effects on fatty acid oxidation. Their abilities to either support, in the absence of carnitine, or inhibit, in the presence of carnitine, carnitine dependent fatty acid oxidation were determined by the conversion of radiolabeled [1-14C]palmitic acid to acid-soluble radiolabeled products. None of the methylcarnitine analogs were observed to be significant inhibitors of palmitate oxidation at concentrations (1.0 mM) up to ten times that for L-carnitine. The two diastereomers of D,L-4-methylcarnitine, however, were able to support palmitate oxidation in the absence of carnitine, and rates were roughly 40% of that obtained with equimolar (0.1 mM) L-carnitine. PMID- 8240426 TI - Do infections trigger fibromyalgia? PMID- 8240427 TI - "Chronic Lyme disease" as the incorrect diagnosis in patients with fibromyalgia. AB - OBJECTIVE: To evaluate a large number of patients referred with persistent symptoms thought to represent chronic Lyme disease. METHODS: We retrospectively reviewed the charts of nearly 800 patients referred with persisting nonspecific musculoskeletal and/or neurologic symptoms thought to represent chronic Lyme disease. RESULTS: Seventy-seven patients were found to have fibromyalgia, not ongoing Lyme disease, as the explanation of their chronic symptoms. Many had received multiple courses of antibiotic therapy for symptoms of fibromyalgia mistakenly attributed to chronic Lyme disease. No patient reported permanent and/or total resolution of fibromyalgia symptoms following antibiotic therapy. Appropriate therapy for fibromyalgia in those who remained compliant, however, was often effective in improving some if not all of the chronic symptoms. CONCLUSION: Fibromyalgia is a treatable and potentially curable disorder, and should be considered in the evaluation of patients with "refractory Lyme disease." PMID- 8240428 TI - The effect of rheumatoid arthritis and steroid therapy on bone density in postmenopausal women. AB - OBJECTIVE: To assess bone mineral density (BMD) in postmenopausal women with rheumatoid arthritis (RA) and the relative effects of disease activity, disability, and past and current use of corticosteroids. METHODS: One hundred ninety-five postmenopausal patients with RA were compared with 597 post menopausal control subjects. Bone density was measured at the lumbar spine and the proximal femur using dual x-ray absorptiometry. Patients were divided into 3 groups according to corticosteroid use, i.e., never users (61%), current users (21%), and ex-users (18%). RESULTS: Compared with controls, the never users had no difference in BMD at the lumbar spine, but a 6.9% reduction at the femur (95% confidence interval [95% CI] 3.4-10.3%). In current users (mean daily prednisolone dosage 6.9 mg), BMD was reduced by 6.5% at the spine (95% CI 0 13.0%) and by 7.4% at the hip (95% CI 1.2-13.6%) compared with never users, after adjustment for age, weight, duration of menopause, and functional disability. Mean BMD was similar in the ex-user and never user groups. Results were confirmed in 54 patients who had whole-body BMD measurements. There were inverse correlations between BMD and Health Assessment Questionnaire scores (femoral BMD r = -0.23, P < 0.01; whole-body BMD r = -0.40, P < 0.01) and between BMD and cumulative steroid dose (femoral BMD r = -0.32, P < 0.01; whole-body BMD r = 0.72, P < 0.01). CONCLUSION: Osteoporosis in postmenopausal women with RA is more evident at the hip than the spine, and the most important determinants of bone loss are disability and cumulative corticosteroid dose. Low-dose steroids cannot be used with complacency, but recovery after discontinuation of use may be possible. PMID- 8240429 TI - Regulation of human normal and osteoarthritic chondrocyte interleukin-1 receptor by antirheumatic drugs. AB - OBJECTIVE: To determine the effect of antirheumatic drugs and corticosteroids on interleukin-1 receptor (IL-1R) levels in, and IL-1-stimulated metalloprotease synthesis and expression by, normal and osteoarthritic (OA) human articular chondrocytes. METHODS: IL-1R affinity and density of human chondrocytes were determined using radioligand binding experiments. Collagenase and stromelysin synthesis activities were analyzed by 14C-labeled type I collagen and Azocoll assays, respectively. Their messenger RNA (mRNA) levels were determined by Northern blot analysis. IL-1 alpha, IL-1 beta, IL-1 receptor antagonist, and beta 2-microglobulin were measured using enzyme-linked immunosorbent assays. Protein synthesis was determined by 3H-leucine incorporation. RESULTS: Antirheumatic drugs reduced the IL-1R level in normal and OA chondrocytes in a dose-dependent manner. In normal chondrocytes, tenidap reduced the IL-1R level by 44% at 5 micrograms/ml to 88% at 100 micrograms/ml (50% inhibition constant [IC50] 10.1 micrograms/ml), indomethacin reduced IL-1R by 6% at 1.5 micrograms/ml to 43% at 60 micrograms/ml, and naproxen reduced IL-1R by 10% at 10 micrograms/ml to 41% at 300 micrograms/ml; the effects observed with indomethacin and naproxen occurred only when the drugs were used at levels above their therapeutic concentrations. In OA chondrocytes, the effect of indomethacin and naproxen on the IL-1R level was greatly reduced, whereas tenidap still had a marked effect (IC50 22.5 micrograms/ml). Dexamethasone and hydrocortisone had no consistent effect on the IL-1R level. At a therapeutic concentration (20 micrograms/ml), tenidap was found to reduce the IL-1R level in a time-dependent manner, with maximum inhibition (98%) by 48 hours. Tenidap was also found to markedly reduce collagenase and stromelysin synthesis and mRNA levels in IL-1-stimulated chondrocytes. CONCLUSION: The suppressive effects of tenidap on IL-1-stimulated metalloprotease synthesis and expression in OA and normal chondrocytes are likely related to a decrease in IL-1R levels. At therapeutic concentrations, tenidap has a greater effect on the IL-1R level than is seen with indomethacin or naproxen, and glucocorticoids have no effect on IL-1R. PMID- 8240430 TI - Differential in vivo expression of collagenase messenger RNA in synovium and cartilage. Quantitative comparison with stromelysin messenger RNA levels in human rheumatoid arthritis and osteoarthritis patients and in two animal models of acute inflammatory arthritis. AB - OBJECTIVE: To compare quantitatively the in vivo expression of collagenase messenger RNA (mRNA) and stromelysin mRNA in the joint tissues of human osteoarthritis (OA) and rheumatoid arthritis (RA) patients and in two animal models of acute inflammatory arthritis. METHODS: In vivo levels of metalloproteinase mRNA and protein were determined by quantitative Northern hybridization and by enzyme-linked immunosorbent assay, respectively. RESULTS: In synovium, mean levels of collagenase mRNA were similar to those of stromelysin mRNA; however, in cartilage, mean levels of collagenase mRNA were significantly lower. The ratios of collagenase mRNA to stromelysin mRNA levels in RA and OA cartilage reflected similar ratios of collagenase protein to stromelysin protein levels in synovial fluid. CONCLUSION: The regulation of collagenase mRNA expression in cartilage is distinct from that of stromelysin, suggesting distinct roles for these two metallo-proteinases in normal and abnormal physiologic functioning of cartilage. PMID- 8240431 TI - A Canadian study of the total medical costs for patients with systemic lupus erythematosus and the predictors of costs. AB - OBJECTIVE: We conducted a cost identification analysis on 164 consecutive patients with systemic lupus erythematosus (SLE) who entered the Montreal General Hospital Lupus Registry between January 1977 and January 1990, compared their costs to the population of Quebec, and determined the predictors of cost. METHODS: In January 1990 and 1991, participants completed questionnaires on health services utilization and on employment history over the preceding 6 months, as well as on functional, psychological, and social well-being. The societal burden of SLE was determined in terms of direct costs (all resources consumed in patient care) and indirect costs (wages lost due to lack of work force participation because of morbidity). RESULTS: The mean total annual cost for 1989, as assessed in January 1990 and expressed in 1990 Canadian dollars, was $13,094. Although only 44% of the patients were fully employed, indirect costs were responsible for 54% of this total ($7,071). Ambulatory costs, primarily diagnostic procedures, medications, and visits to health care professionals, comprised 55% of direct costs ($3,331). The results of the 1990 cost determination were similar. On average, hospitalizations among SLE patients were 4 times more frequent than among the general population of Quebec (matched for age and sex), and the number of ambulatory visits to physicians was double that for the average resident of Quebec. Higher 1989 values of creatinine and a poorer level of physical functioning were the best predictors of higher 1990 direct costs (R2 = 0.29). A poorer SLE well-being score, a combination of education and employment status, and a weaker level of social support were the best predictors of higher indirect costs (R2 = 0.29). CONCLUSION: The direct and indirect costs for patients with SLE are substantial, and their respective predictors are distinct. Direct costs arise from organic complications which induce functional disability. Predictors of indirect costs are potentially amenable to psychological or social interventions and may be more easily modified than the determinants of direct costs, thereby improving patient outcome while simultaneously reducing disease costs. PMID- 8240432 TI - A distinctive autoantibody profile in black female patients with lupus nephritis. AB - OBJECTIVE: Lupus nephritis has often been associated with anti-DNA, but, based on the findings in eluate studies, it appears that other antigen-antibody reactions, such as those involving anti-Ro/SS-A, anti-nuclear RNP (anti-nRNP), and/or anti Sm, may also contribute to the pathogenesis of nephritis. In the present investigation, we identified and further studied a distinctive precipitin profile present in black women with nephritis. METHODS: Longitudinal clinical and serologic studies of a cohort of university-based systemic lupus erythematosus (SLE) patients (n = 120) were carried out over an 8-year period. RESULTS: A subset of 20 black female patients was identified, of whom 8 had lupus nephritis (group I) and 12 did not (group II). Group I was characterized by a distinct precipitin profile consisting of anti-Ro/SS-A, anti-SM, and anti-nRNP, but no anti-La/SS-B. SLE disease duration at presentation was significantly shorter in group I than in group II (mean 1.94 years versus 5.21 years; P = 0.02). The distinctive precipitin profile of anti-Ro/SS-A, anti-Sm, and anti-nRNP occurred exclusively in group I patients (6 of 8, versus 0 of 12 in group II; P < 0.001). In white lupus nephritis patients, this precipitin profile was not seen. CONCLUSION: While the mechanism responsible for the relationship of this distinctive serologic profile to the development of nephritis in black female lupus patients remains to be determined, its presence may be used as a marker for severe and progressive renal disease. PMID- 8240433 TI - Low levels of total cholesterol, high-density lipoprotein, and apolipoprotein A1 in association with anticardiolipin antibodies in patients with systemic lupus erythematosus. AB - OBJECTIVE: To determine if there is an association between low levels of high density lipoprotein cholesterol (HDL), apolipoprotein A1 (Apo A1), total cholesterol, and anticardiolipin antibody (aCL) in patients with systemic lupus erythematosus (SLE) who are not taking corticosteroids. METHODS: We studied 75 outpatients with documented SLE who were attending our hospital clinics: 57 were aCL positive and 18 were aCL negative. Both IgG and IgM aCL levels were determined by enzyme-linked immunosorbent assay. Lipid fractions (total cholesterol, HDL, low-density lipoprotein, very-low-density lipoprotein, and triglycerides) were determined by standard enzymatic techniques. Apo A1 and Apo B levels were determined by nephelometry. RESULTS: Patients with SLE who were IgG aCL+ had low levels of serum cholesterol (mean +/- SD 173.6 +/- 34.6 mg/dl) and HDL (43.9 +/- 16.3 mg/dl) compared with aCL- SLE patients, normal donors, and patients with other diseases. Apo A1 levels were also low in the aCL+ group (95.5 +/- 50.9 mg/dl) compared with the aCL- group (152.7 +/- 32.6 mg/dl). There was no association of total cholesterol level or aCL titer with clinical activity. CONCLUSION: These data indicate that in SLE patients, there is an association between antibody against the phospholipid cardiolipin and low levels of cholesterol, HDL, and Apo A1. PMID- 8240434 TI - The course of skin involvement in systemic sclerosis over three years in a trial of chlorambucil versus placebo. AB - OBJECTIVE: To describe the course of cutaneous involvement in systemic sclerosis (SSc; scleroderma) over 3 years, in the context of a placebo-controlled drug trial. METHODS: The course of skin tethering (assessed by a semiquantitative skin scoring technique) was documented annually for 3 years in 64 SSc patients with early (< 3 years duration at entry), intermediate (3-8 years duration), or late (> 8 years duration) diffuse or limited SSc. RESULTS: Mean +/- SD entry skin scores were significantly greater in the 33 diffuse SSc patients (13.1 +/- 5.0) than in the 31 limited SSc patients (4.3 +/- 2.1) (P < 0.001). In patients with diffuse SSc, the skin score remained stable for the first 12 months, but had decreased significantly by 24 months (P < 0.022) and 36 months (P < 0.004). In those with limited SSc, the skin score did not change significantly over 3 years. CONCLUSION: Trials of treatments designed to affect skin thickening/tethering should be conducted in patients who have diffuse SSc (of short, intermediate, or long duration) at entry. The best time to study therapies designed to affect skin thickening may be in the first year after entry. PMID- 8240435 TI - Isolated congenital heart block. Long-term outcome of mothers and characterization of the immune response to SS-A/Ro and to SS-B/La. AB - OBJECTIVE: To study the long-term outcome of mothers of children with isolated congenital heart block (CHB) and to characterize the maternal autoantibody response to SS-A/Ro and SS-B/La. METHODS: A retrospective clinical study of 33 mothers a mean of 11.2 years (SD 9.2 years, range 0-32 years) after the delivery of their first child with CHB. A clinical and immunologic study of 31 of these mothers, compared with 89 healthy mothers, 45 mothers with systemic lupus erythematosus (SLE), and 19 mothers with primary Sjogren's syndrome (SS), all of whom had healthy children. The specificity of the autoantibody responses to SS A/Ro and SS-B/La was studied by enzyme-linked immunosorbent assays using purified human recombinant antigens and affinity-purified antigens. RESULTS: By the time of the analysis, 2 (6%) of the 33 mothers of CHB children had died and 6 (18%) had met the criteria for SLE. As a group, mothers of CHB children had clinical and immunologic characteristics more closely related to primary SS than to SLE or any other connective tissue disease. The predominant autoantibody response was to the SS-A/Ro antigens, notably to the 52-kd SS-A/Ro protein (prevalence 97%). Compared with controls with SLE, mothers of CHB children had higher titers of antibodies to recombinant 52-kd and 60-kd SS-A/Ro proteins and to the affinity purified SS-A/Ro antigen (P < 0.05, P < 0.01, and P < 0.001, respectively). Compared with controls with primary SS, the autoantibody responses were similar. CONCLUSION: The predominant autoimmune disorder in mothers of children with CHB is subclinical primary SS. Antibodies to SS-A/Ro appear to be a prerequisite for the development of CHB. PMID- 8240436 TI - U1-70-kd autoantibody-positive mixed connective tissue disease in children. A longitudinal clinical and serologic analysis. AB - OBJECTIVE: To obtain longitudinal data on the clinical, serologic, and immunogenetic features of children with mixed connective tissue disease (MCTD). METHODS: Eleven children with MCTD were followed up for a mean of 9.8 years. Enzyme-linked immunosorbent assay and immunoblotting were used to analyze sera for autoantibodies to small nuclear ribonucleoprotein polypeptides. HLA types were determined in 9 patients, by microcytotoxicity and DNA typing. RESULTS: All 11 children had anti-U1-70-kd auto-antibodies. Six of 9 were positive for HLA DR2, 4 of 9 for HLA-DR4, and 9 of 9 for either HLA-DR2 or DR4. Outcomes were favorable with no functional impairment in 8 of the 11 children and were poor in 3. CONCLUSION: The frequency of HLA-DR2/DR4 is increased among children with anti U1-70-kd autoantibody positive MCTD. PMID- 8240438 TI - Inflammatory polyarthritis in mice transgenic for human T cell leukemia virus type I. AB - OBJECTIVE: We have recently reported that arthropathy develops in high incidence among transgenic mice carrying the pX region of human T cell leukemia virus type I (HTLV-I). In the present study, the histopathologic features of the joints in these mice were examined in order to compare the animal disease with rheumatoid arthritis (RA) in humans. METHODS: Paraffin sections of limbs (right and left fingers, wrists, elbows, shoulders, toes, knees, and ankles) were stained with hematoxylin and eosin, periodic acid-Schiff, azan-Mallory, or phosphotungstic acid hematoxylin, and examined by light microscopy. RESULTS: Abnormalities of the limbs began to occur as early as 3 weeks of age, and the incidence gradually increased until the mice were 12 months old. The incidence of arthropathy was 22% (48 of 217) at 3 months of age and 28% (18 of 64) at 6 months. The severity of the histopathologic changes in the joints of the transgenic mice ranged from grade I to grade IV. CONCLUSION: The major histopathologic features in the joints of HTLV-I transgenic mice are similar to those in humans with RA. Thus, these mice may represent a useful model for the study of the disease in humans. PMID- 8240437 TI - Phosphocitrate prevents disease progression in murine progressive ankylosis. AB - OBJECTIVE: Mice with progressive ankylosis, a spontaneous arthropathy, were treated with phosphocitrate (PC) in vivo to determine the effect of PC on disease progression. METHODS: Two groups of mice with progressive ankylosis (matched for age, weight, and sex) were treated parenterally for 6 weeks with either PC or saline vehicle. RESULTS: Clinically, histologically, and microradiographically, there were significant differences in disease progression and severity in the PC treated and the saline-treated mice. CONCLUSION: PC appears to inhibit disease progression in murine progressive ankylosis. PMID- 8240439 TI - Persistence of Borrelia burgdorferi in ligamentous tissue from a patient with chronic Lyme borreliosis. AB - OBJECTIVE: To document the persistence of Borrelia burgdorferi in ligamentous tissue samples obtained from a woman with chronic Lyme borreliosis. METHODS: Spirochetes were isolated from samples of ligamentous tissue, and the spirochetes were characterized antigenetically and by molecular biology techniques. The ligamentous tissue was examined by electron microscopy. Humoral and cellular immune responses were analyzed. RESULTS: Choroiditis was the first recognized manifestation of Lyme disease in this patient. Despite antibiotic therapy, there was progression to a chronic stage, with multisystem manifestations. The initially significant immune system activation was followed by a loss of the specific humoral immune response and a decrease in the cellular immune response to B burgdorferi over the course of the disease. "Trigger finger" developed, and a portion of the flexor retinaculum obtained at surgery was cultured. Viable spirochetes were identified. Ultramorphologically, the spirochetes were situated between collagen fibers and along fibroblasts, some of which were deeply invaginated by these organisms. The cultured bacteria were identified as B burgdorferi by reactions with specific immune sera and monoclonal antibodies, and by polymerase chain reaction amplification and Southern blot hybridization techniques. CONCLUSION: To our knowledge, this is the first report of the isolation of B burgdorferi from ligamentous tissue. This suggests that tendon tissues serve as a specific site of spirochete residence in human hosts. PMID- 8240440 TI - Bronchiolitis obliterans organizing pneumonia associated with essential mixed cryoglobulinemia. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) is an uncommon and underrecognized pathologic finding observed in a variety of syndromes including connective tissue diseases. Although lung involvement in essential mixed cryoglobulinemia (EMC) has been reported, there are no previous reports of BOOP in EMC. We report herein a case of longstanding EMC associated with interstitial and cavitary lung disease histologically consistent with BOOP. A theory of a possible etiologic basis for the development of BOOP in patients with connective tissue diseases is discussed. PMID- 8240441 TI - Mycobacterium kansasii septic arthritis in a patient with acquired immune deficiency syndrome. PMID- 8240442 TI - Sterile oily abscess from depot gold therapy. PMID- 8240443 TI - Bacterial antibodies in HLA-B27+ healthy individuals. PMID- 8240444 TI - The hazards of gold injections in children with juvenile rheumatoid arthritis: comment on the article by Lehman. PMID- 8240445 TI - Rheumatoid arthritis, fecundity, and coital rates: comment on the article by Nelson et al. PMID- 8240446 TI - Treatment of asymptomatic, retrospectively diagnosed, Lyme disease: comment on the report by Christian. PMID- 8240447 TI - In vivo percutaneous absorption of [14C]acitretin in the hairless guinea pig and in the rhesus monkey. AB - The oral administration of retinoids such as etretinate and acitretin (Ro 10 1670, CAS 55079-83-9), provides a successful therapeutic approach in various cutaneous diseases characterized by disturbed keratinization, e.g. psoriasis. Nevertheless oral therapy is often associated with systemic adverse effects. This makes a topical form with no or reduced systemic side effects desirable. Direct application of a topical acitretin formulation to the skin might result in therapeutic skin concentrations while minimizing systemic exposure. In the hairless guinea pig and in the rhesus monkey the percutaneous absorption of [14C]acitretin from an isopropylmyristate formulation (160 micrograms acitretin/2.5 cm2/animal) were investigated in vivo. After a 24 h exposure drug concentration in the skin was higher in the hairless guinea pig (620 ng-eq/g wet tissue) than in the rhesus monkey (380 ng-eq/g wet tissue). A similar observation was made comparing the 24 h absorption data determined as amount of drug excreted. The results are compared with in vitro absorption data using skin from the same species. PMID- 8240448 TI - Design, synthesis and biological study of new antiparasitic spiroarsoranes. AB - Thirty-eight new spiroarsoranes were synthesized after structure-activity relationship studies from the first series. These compounds were predicted to cross more easily the membrane of protozoae or the cuticle of Nematodes and to reach their biological target with efficiency. The spiroarsoranes were evaluated for their antiparasitic properties, on helminths and protozoa models in regard of their parent arsonic acids. The following parasite models were used: Entamoeba histolytica and Trichomonas vaginalis in vitro; Molinema dessetae infective larvae in vitro, adults and microfilariae in vivo; Nippostrongylus brasiliensis infective larvae in vitro. The results obtained on these models indicated that the "spiranization" of arsonic acids produced new compounds with a biological activity 10-fold superior to those of arsonic acids. Nevertheless, each parasite had its own sensitivity to spiroarsoranes. Moreover, in vivo results showed that the lipophilicity of the molecules should be optimal to avoid high toxicity in host such as arsenical encephalopathy. PMID- 8240449 TI - In vitro action of new organometallic compounds against Trypanosomatidae protozoa. AB - The effects of 10 newly synthesized organometallic rhodium complexes (5 of Rh(I), 4 of Rh(III) and one of Rh(IV) complex salt) and 2 antimony (III) complexes against epimastigote forms of Trypanosoma cruzi and promastigote forms of Leishmania donovani have been studied in vitro. Of the 10 rhodium complexes tested three proved 100% efficient in inhibiting the growth of the epimastigote forms of T. cruzi while only one worked against the promastigote forms of L. donovani. The two antimony complexes inhibited the growth of L. donovani totally and were fairly successful against T. cruzi. PMID- 8240450 TI - Twice daily dosing of erythromycin acistrate in the treatment of acute bronchitis and pneumonia. AB - Erythromycin acistrate (Erasis, CAS 96128-89-1) is a 2'-acetyl ester prodrug of erythromycin. Due to prolonged half-life it is more suitable for twice daily dosing than the conventional erythromycin preparations. In the present double blind trial, totally 297 ambulatory patients with respiratory tract infections were treated either with erythromycin acistrate 800 mg daily or doxycycline 100 mg daily. 243 of the included patients had bronchitis, 15 patients bronchitis and other respiratory tract symptoms, 25 patients pneumonia and 14 other respiratory tract infections. The duration of treatment varied from 7 to 14 days depending on the severity of infection. The efficacy of both treatments was very good. 96.6% of the patients treated with erythromycin acistrate and 97.2% of the patients treated with doxycycline improved. The efficacy of erythromycin acistrate in the treatment of bronchitis and pneumonia was 96.7 and 100%, respectively. Only 5 of the totally 148 patients failed. Side effects (mainly gastrointestinal symptoms) were seen in 12.1% of the patients (20 patients in the erythromycin acistrate group and 16 patients in the doxycycline group). The results show that erythromycin acistrate dosed twice daily is as effective as doxycycline and well tolerated in the treatment of lower respiratory tract infections. PMID- 8240451 TI - Effects of the combination of ketoconazole and calmodulin inhibitors against Candida albicans in vitro. Short communication. AB - The susceptibility of 66 strains of Candida albicans from patients was tested against ketoconazole (Ktz), chlorpromazine (Chl) levomepromazine (Lev), haloperidol (Hal) and the combination of Ktz with these calmodulin inhibitors, using Sabouraud's broth. The minimal inhibitory concentrations (MICs) for 66 strains of C. albicans were as follows: Chl 192 +/- 11.4 micrograms/ml, Lev 306 +/- 16.4 micrograms/ml, Hal 464 +/- 13.8 micrograms/ml compared with Ktz 34.46 +/ 3.9 micrograms/ml. The combination of Ktz and calmodulin inhibitors in various ratios (1:1,1:2,2:1) was found to exert synergistic effect and the mean values of the combinations were: Ktz+Chl 3.45 +/- 0.35, 3.78 + 0.36, 5.58 + 0.4 micrograms/ml; Ktz+Lev 10.8 +/- 2.19, 9.7 +/- 2.23, 10.5 + 2 micrograms/ml; Ktz+Hal 6.4 +/- 1.7, 6.8 +/- 1.6, 7.28 +/- 1.5 micrograms/ml. These results were significantly different (p < 0.001) from those of ketoconazole. These findings indicate that some calmodulin inhibitors increase the antifungal activity of Ktz against C. albicans in vitro. PMID- 8240452 TI - Nonlinear pharmacokinetic modeling of a gadolinium chelate used as a liver specific contrast agent for magnetic resonance imaging. AB - The introduction of the lipophilic moiety, ethoxybenzyl, into the gadolinium chelate gadopentate dimeglumine (Gd-DTPA, Magnevist, CAS 86050-77-3) yielded gadolinium-(4S)4-(4-ethoxybenzyl)-3,6,9-tris(carboxylatomethyl+ ++)-3,6,9- triazaundecandioic acid, disodium salt (Gd-EOB-DTPA) a compound with a potential as a magnetic resonance contrast agent for liver mass screening. After intravenous administration in rats (0.05 and 0.5 mmol/kg), dogs (0.03 and 0.25 mmol/kg) and monkeys (0.25 mmol/kg) Gd-EOB-DTPA was only slightly bound to plasma proteins and underwent both renal and extrarenal elimination. The pharmacokinetic behaviour in rats, dogs and monkeys could be well described by disposition of the compound in the central and peripheral compartment with elimination occurring from the central compartment, allowing for Michaelis-Menten kinetics for the extrarenal (biliary) route of elimination and linear kinetics for the renal route of elimination. Nonlinear pharmacokinetic modeling yielded Vmax values (mumol/min.kg) of 4.34 +/- 0.65 in rats, 2.40 +/- 0.98 in dogs and 0.810 +/- 0.66 in monkeys. In the rat the Vmax value obtained by nonlinear modeling was in good agreement with the biliary transport maximum measured after intravenous bolus injection of increasing dose levels. PMID- 8240453 TI - Some biochemical properties of the new potential antidepressant agent N-methyl-N propargyl-2-(1-methyl-5-methoxyindolyl)methylamine. AB - The biochemical properties of the new methyl indole derivative IM-24 (N-methyl-N propargyl-2(1-methyl-5-methoxyindolyl)methylamine HCl) have been investigated. The activity on both forms of monoamine oxidase MAO was tested in several nervous and non nervous tissues ex vivo after chronic administration. IM-24 is mainly an inhibitor of the activity of MAO A without any effect on intestinal MAO B at the doses studied. IM-24 was compared with tricyclic antidepressants in tests for serotonin (5HT), noradrenaline (NA) and dopamine (DA) uptake inhibition in vitro. IM-24 is mainly an inhibitor of the 5HT uptake mechanism but is less active than paroxetine and chlorimipramine which are very potent 5HT-uptake inhibitors. Radioligand binding techniques in rat brain ex vivo showed that IM-24 after chronic administration (21 days) produces no change in the number or the affinity of the alpha 2-adrenoceptors. IM-24 reduces by 70% the number of 5HT2 receptors but does not modify the affinity for the ligand. IM-24 is thus an interesting compound which combines monoamine oxidase inhibition with inhibition of 5HT uptake. Both these actions will lead to an increase of the availability of serotonin at the synaptic site. PMID- 8240454 TI - Suppression of opiate withdrawal and cocaine hyperarousal syndromes by buspirone. Possible pharmacotherapeutic applications. AB - Buspirone (CAS 36505-84-7) was evaluated in three animal models which were designed to study stages of drug abuse most likely associated with compulsive abuse. Buspirone attenuated abrupt withdrawal in rhesus monkeys maximally dependent on morphine. In addition, it completely blocked the emergence of cocaine-induced stereotyped behavior in rats and attenuated the hyperarousal or rausch syndrome in morphine-dependent and non-dependent rhesus monkeys. Buspirone was active at doses which caused little, if any, impairment in the animals. The results suggest that buspirone may possibly find application in the pharmacotherapy of opioid and cocaine abuse. PMID- 8240455 TI - Protective effect of crataegus extract on the cardiac mechanical dysfunction in isolated perfused working rat heart. AB - The effect of the water-soluble fraction of Crataegus (Crataegus extract) on the cardiac mechanical and metabolic function was studied in the isolated, perfused working rat heart during ischemia and reperfusion. Ischemia (15 min) was produced by removing afterload pressure, and reperfusion (20 min) was produced by returning it to the original pressure. In the control (no drug) heart, ischemia decreased mechanical function to the lowest level, which did not recover even after the end of reperfusion. Crataegus extract (0.01 or 0.05%) was applied to the heart from 5 min before ischemia through the first 10 min after reperfusion. With the high concentration of Crataegus extract (0.05%) the mechanical function recovered during reperfusion incompletely without increasing coronary flow, but the low concentration of Crataegus extract (0.01%) did not. In the heart treated with the high concentration of Crataegus extract, the reperfusion-induced recovery of the energy metabolism was accelerated, and the level of lactate during ischemia was lower than that in the control heart, although the myocardial levels of free fatty acids during ischemia and reperfusion were not greatly affected. These results demonstrate that Crataegus extract (0.05%) has a cardioprotective effect on the ischemic-reperfused heart, and that the cardioprotective effect is not accompanied by an increase in coronary flow. PMID- 8240456 TI - Effects of inosine on adenosine-induced coronary vasodilation in the open chest dog. AB - The effects of inosine (CAS 58-63-9) on adenosine-induced coronary vasodilation were studied in open-chest dogs. Inosine and hypoxanthine were infused into the coronary artery at a rate to obtain respective calculated coronary plasma concentrations of 10(-5) mol/l, and the dose-coronary flow response of adenosine was recorded with and without inosine or hypoxanthine infusion. When the maximum coronary dilation was obtained, 10 ml of 2 x 10(-3) mol/l 8-phenyltheophylline (8 PT) solution was injected into the femoral vein. Additionally, adenosine deaminase activity was measured in vitro in the presence of various concentrations of either inosine or hypoxanthine. It was found that inosine, but not hypoxanthine, intensified the coronary vasodilatory effect of adenosine, which was abolished by 8-PT injection: EC50 of adenosine was reduced from 10( 5.43) mol/l to 10(-5.90) mol/l by inosine. Inosine and hypoxanthine did not affect adenosine deaminase activity at concentrations of 10(-4) mol/l or less. These findings indicate that inosine intensifies the coronary vasoactivity of adenosine, independent of inhibition of adenosine deaminase activity. PMID- 8240457 TI - Pharmacokinetics of dilevalol and its conjugates in man. Assay method for plasma, blood, urine and bile samples and preliminary pharmacokinetic studies. AB - The renal and biliary excretion of the beta-adrenoceptor blocking agent dilevalol (CAS 75659-07-3) and its conjugates was examined in a preliminary pharmacokinetic study. Plasma, urine and bile dilevalol concentrations were determined with a simplified procedure that is based on alkaline liquid-liquid extraction using diethyl ether and subsequent reversed-phase HPLC separation of the reconstituted samples (on a PRP-1 stationary phase using a mixture of methanol and pH 9.8 carbonate buffer as mobile phase). Triamterene was used as internal standard. The quantification of the conjugates was accomplished indirectly via enzymatic hydrolysis (glusulase) with and without addition of the beta-glucuronidase inhibitor 1,4-saccharolactone (at a final concentration of 5.5 mmol/l). In the pharmacokinetic study healthy volunteers and cholecystectomised patients with a T drain received a single oral dose of 200 mg dilevalol. Furthermore, to healthy volunteers an i.v. dose of 60 mg dilevalol was given in order to estimate the absolute bioavailability. From the obtained data the systemic plasma clearance was calculated to be 1708 ml/min. The oral bioavailability was calculated to be 16%. The log concentration-time curves of the metabolites paralleled those of dilevalol in the terminal section with average terminal half-lives of approx. 5 h. In volunteers the fractions of the dose excreted renally were 0.5% for parent drug, 23% for the glucuronide(s) and 8% for the sulfate. The corresponding values found for the patients were not significantly different. In the patients' bile only 1.2% of the total dose were found (0.03% dilevalol, 1.1% dilevalol glucuronide(s), 0.1% dilevalol sulfate).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240458 TI - [Circadian antihypertensive effect of sustained-release isradipine in patients with essential hypertension in comparison to placebo]. AB - In this multicenter, placebo-controlled study, 16 patients with mild to moderate essential hypertension were treated with 10 mg/day isradipine retard (PN 200-110, Lomir SRO, CAS 75695-93-1) for 3 weeks. The study started with a 2 week placebo wash out phase. 13 patients were randomised to an exclusive placebo therapy. After the placebo wash out phase, following the 1st medication in active therapy and after the end of therapy, 24-h blood pressure profiles were recorded. The profile under placebo on the 1st medication was separated by a one-week intervening placebo therapy for all patients. On active therapy, the systolic as well as the diastolic blood pressure (day time, night time and 24-h mean values) were significantly reduced. The antihypertensive effect of the active therapy became already manifest after the 1st medication and was augmented after 3 weeks of therapy. In the placebo group no parameter of the 24-h profiles changed significantly. The tolerability of treatment was excellent in 14 (87.5%) of the isradipine patients and in 10 (76.9%) of the placebo group. In one of 16 patients in the active group, adverse events (flush and ankle oedema) were observed. However, therapy could be continued. In one patient of the placebo group, oedema of the fingers was noticed, in another headache was documented. In the placebo group two patients discontinued the study due to inefficacy, in the isradipine group one patient for the same reason; a second patient was excluded from this group due to a concomitant disease unrelated to the study drug. PMID- 8240459 TI - Disposition characteristics of coumarin as a function of age in the beagle dog model. AB - Coumarin (C, CAS 91-64-5) is a high extraction ratio drug, and its clearance reflects the liver blood flow. The purpose of this study was to investigate the alterations in the pharmacokinetics of C as a function of age, and to evaluate the suitability of beagle dog model to study age dependent changes in the hepatic function. In this longitudinal study, C was administered intravenously to male beagle dogs, ranging in age from < 1 to 15 years. The total clearance decreased significantly from 30.67 +/- 9.58 to 16.61 +/- 2.35 ml/min/kg and the ratio of AUC of glucuronide metabolite to parent drug decreased from 1.26 +/- 0.52 to 0.72 +/- 0.17 in old dogs. No significant changes were observed in apparent volume of distribution and elimination half-life. The changes in clearance of C parallels the observed reduction in the liver blood flow in man, suggesting that the beagle dog appears to be a good model for drug disposition studies in gerontokinetics. PMID- 8240460 TI - Pharmacokinetics of estradiol, free and total estrone, in young women following single intravenous and oral administration of 17 beta-estradiol. AB - The pharmacokinetic parameters of estradiol (E2, CAS 50-28-2), free and total estrone (E1, CAS 53-16-7) were determined in 14 young women following a single oral administration of 2, 4 and 8 mg E2 and a single intravenous administration of 0.3 mg E2 in an open, intraindividual comparison with 4 treatments. The purpose of the study was to determine the absolute bioavailability of orally administered E2 in a larger group of women and to assess the inter- and intraindividual variability of basic pharmacokinetic parameters of E2 and metabolically derived E1. In addition, the outcome of this study should provide a basis for the decision whether E2 could potentially be used in a combination oral contraceptive. There was a dose proportional increase in the AUC-values following the oral administration of 2 mg and 4 mg doses of E2. At the high dose of 8 mg, however, only about 76%, 78% and 70% of the expected values were found for E2, free and total E1, respectively. Especially the reduction in total E1 concentrations points to an incomplete absorption of E2 at the high dose level. The absolute bioavailability of orally administered E2 was calculated based on the 4 mg dose and was found to be 4.9 +/- 5.0%. The mean ratio of free E1 and E2 concentrations in the serum, following parenteral and oral administration of E2 was about 1.0 (i.v.) and between 8.8 to 19.8 (p.o.), respectively. Pharmacokinetic parameters, like AUC, derived from serum level-time curves of E2, free and total E1 showed a high intra- and interindividual variability.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240461 TI - Pharmacokinetics and organ distribution of liposome-encapsulated L-carnitine in rats. AB - The pharmacokinetics and organ distribution of 3H-L-carnitine (CAS 541-15-1) were investigated in rats following direct intravenous administration of the drug substance and administration of the drug encapsulated in liposomes, respectively. The retention in the blood system of carnitine in liposomes, was significantly higher, namely up to 300% as compared to the standard administration. The half life of distribution t1/2 alpha to 0.68 h (+154%), the terminal half-life t1/2 beta to 7.94 h (+140%), whereas the total clearance decreased by 400% as compared with the standard carnitine administration. Carnitine, in the novel dosage form, accumulated to a higher extent in the liver (156%) and spleen (336%), while the concentration in lung (52%), heart (55%) and muscle tissue (54%) decreased markedly relative to the standard. The novel dosage form is stable in vitro (t1/2 4 degree C = 187 days) as well as in vivo and, thus, may be successfully used in the therapy of carnitine deficiency, for instance in patients with renal failure or liver disease. PMID- 8240462 TI - Limitation of the deterioration of lipid parameters by a standardized garlic ginkgo combination product. A multicenter placebo-controlled double-blind study. AB - The efficacy of a garlic-ginkgo combination product (Allium plus) was analyzed in a randomized placebo-controlled double-blind study under extreme dietary conditions. The Christmas/New Year's season was chosen for this 2 months lasting investigation analyzing whether the known cholesterol lowering effect of garlic was even effective during the period of the year with the most cholesterol-rich meals. 43 patients with elevated total cholesterol levels ranging between 230-390 mg/dl completed the study. There were no significant changes of the total cholesterol values in both treatment groups. Nevertheless the analysis of improvement or deterioration of total cholesterol values revealed a clear difference between verum and placebo. 20% of the patients in the placebo group showed an improvement of their total cholesterol level, while there was a significant greater improvement rate of 35% in the verum group (p < 0.05). The responders of the verum group showed a reduction in the total cholesterol values from 298.5 +/- 53.8 to 293.0 +/- 56.4 mg/dl after 1 month and a total reduction of 10.4% after 2 months to 267.6 +/- 44.4 mg/dl. The difference after 2 months of treatment was significantly different from the starting value (p < 0.05). After the 2 months treatment phase there was a 2 weeks wash-out period. During this period the total cholesterol value returned to 293.5 +/- 90.1 mg/dl showing the effectiveness of garlic treatment, but indicating the need for a continuous long term therapy. PMID- 8240463 TI - Protective action of vinpocetine against experimentally induced gastric damage in rats. AB - The efficacy of vinpocetine (CAS 42971-09-5) to prevent gastric mucosal damage induced by several noxious agents and its antisecretory effect were studied in rats. Vinpocetine administered orally or intraperitoneally inhibited the development of gastric lesions induced by 96% ethanol in a dose-dependent way. The highest protective activity was observed when vinpocetine was given intraperitoneally 30 min before ethanol, and its effect was still significant when administered 120 min before ethanol exposure. Oral administration of vincamine also displayed gastroprotective action in this model. Pretreatment with indometacin counteracted the protective action of vinpocetine against ethanol induced damage, suggesting the involvement of a prostaglandin-mediated mechanism. The protective effect of vinpocetine was compared with that of prostaglandin E2, sucralfate, and tripotassium dicitrate bismuthate. The antiulcer activity of vinpocetine was demonstrated also in gastric injury induced by phenylbulazone and in chronic gastric ulcer induced by acetic acid. Histamine-stimulated gastric acid secretion in pylorus-ligated rats was partially inhibited by vinpocetine administered intraduodenally. The activity of vinpocetine established in these experiments is indicative of its potential clinical value as a gastroprotective agent. PMID- 8240464 TI - Pharmacokinetics and bioavailability of metoclopramide nasal spray versus metoclopramide intravenous in healthy volunteers and cancer patients. AB - The kinetics and bioavailability of a new formulation of metoclopramide (CAS 364 62-5) nasal spray (MTC NS) were assessed in two separate studies versus the same drug administered intravenously (MTC IV) according to a balanced-block design where each study subject served as his own control. The first study involved 10 healthy subjects, each receiving metoclopramide NS 20 mg (one 10-mg puff per nostril) and metoclopramide IV 20 mg on two trial days separated by a 7-day washout period. On both occasions, blood samples were obtained at time 0 and at 20, 40, 60, 120, 150, 210, 280 and 360 min of dosing. Metoclopramide concentrations were assayed in plasma by HPLC. The second study involved 10 patients of oncologic domain, scheduled to receive mildly emetic chemotherapy regimes. The experimental design was similar to the one above except that blood was sampled at 0, 20, 40 and 60 min and again at 2, 3, 4, 6, and 8 h of dosing. All healthy subjects completed the trial without experiencing any adverse or untoward events; in the group of cancer patients, one subject dropped out after the nose spray treatment when he was removed to another department. This patient was replaced by another, and included in final data analysis only for the segment of treatment actually received. Metoclopramide kinetics after intravenous dosing were in good agreement with known data for the active substance, with no meaningful differences between healthy subjects and cancer patients. With MTC NS administration there was only a slight but significant difference of Cmax, being lower in the cancer patient group (p < 005).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240465 TI - [Circadian dependency of ibuprofen gastropathy and the protective effect of ranitidine. An endoscopic, controlled double-blind pilot study]. AB - Circadian Dependency of Ibuprofen Gastropathy and Protective Effect of Ranitidine/An endoscopic, controlled double-blind pilot study. In a randomized parallel double-blind study, the gastric and duodenal effects of 600 mg S(+) ibuprofen (CAS 15687-27-1) daily in the presence and absence of 300 mg ranitidine (CAS 66357-35-5) was evaluated in 20 healthy volunteers undergoing upper GI endoscopy. Drugs were taken over a period of 7 days either at 8 a.m. (n = 10) or at 8 p.m. (n = 10). Endoscopic controls were performed at entry and repeated after 7 days of treatment. A damage score according to Lanza et al. was used. At entry both groups showed comparable mucosal damages. 8 a.m.-group: ibuprofen/placebo (stomach) 0.9 +/- 0.1 and 0.0 +/- 0.0 (duodenum); ibuprofen/ranitidine 0.8 +/- 0.1 (stomach) and 0.1 +/- 0.1 (duodenum). 8 p.m. group: ibuprofen/placebo 0.9 +/- 0.1 (stomach) and 0.2 +/- 0.1 (duodenum); ibuprofen/ranitidine 0.9 +/- 0.1 (stomach) and 0.1 +/- 0.1 (duodenum). After 7 days of treatment the lesion score increased in the ibuprofen/placebo-group in the 8 a.m.-group to 3.2 +/- 1.2 (stomach) and to 0.7 +/- 0.5 (duodenum), and in the 8 p.m.-group to 8.4 +/- 1.9 (stomach) and to 2.9 +/- 1.2 (duodenum). The corresponding values in the ibuprofen/ranitidine-group were 1.8 +/- 0.8 (stomach) and 0.1 +/- 0.1 (duodenum) (8 a.m.-group) as well as 5.1 +/- 1.4 (stomach) and 0.1 +/- 0.1 (duodenum) (8 p.m.-group). The difference between the morning and the evening dose of ibuprofen as well as ranitidine protection reached statistical significance when the corresponding data were pooled (p < 0.05). Our data suggest that the gastrolesive effects of S(+)-ibuprofen are dependent of the time of drug ingestion; protection by ranitidine, however, was time-independent. PMID- 8240466 TI - Inhibition of neutrophil function in vitro by nimesulide. Preliminary evidence of an adenosine-mediated mechanism. AB - Nimesulide (CAS 51803-78-2) is a methane sulphoanilide derivative provided with specific anti-inflammatory activity. In human polymorphonuclear leucocytes (PMNs), the activity of nimesulide has been suggested to be based on the inhibition of the oxidative burst. However, the effect of the compound on PMNs function seems to be very complex. In order to give a major insight into the mechanism of action of nimesulide, the effect of the drug was studied in vitro on human PMNs functions, such as free radical generation and enzyme release, and on cytosolic free calcium levels, following the activation with specific stimuli. Moreover, the hypothesis that nimesulide could act by interfering with the adenosine cell receptor system was also evaluated. Nimesulide (1-50 mumol/l showed a dose-dependent inhibitory activity on superoxide anion and chemiluminescence production from PMNs stimulated with the oligopeptide fMLP, the ionophore A23187, and the phorbol ester PMA. Enzyme release was significantly reduced, when fMLP and A23187 represented the stimulating agents, while no effect at all was observed with PMA. Studies with the fluorescent calcium chelating dye FURA 2/AM showed that nimesulide was able to reduce free cytosolic calcium increase produced by fMLP and the ionophore ionomycin. The preincubation of cells with the specific adenosine receptor antagonist theophylline was able to significantly reverse the inhibitory activity of nimesulide, either on free radical production and enzyme release, and on free cytosolic calcium increase sustained by fMLP and the ionophores.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240467 TI - Studies on the anti-phospholipase A2 and anti-inflammatory activities of a uteroglobin fragment and related peptides. AB - The nonapeptide antiflammin P1 (H-Met-Gln-Met-Lys-Lys-Val-Leu-Asp-Ser-OH), its isoAsp8 analogue and the corresponding aminosuccinyl peptide were prepared, characterized and tested for inhibition of phospholipase A2 (PLA2) in vitro and for anti-inflammatory activity in vivo under assay conditions recently recommended. All peptides are devoid of PLA2 inhibitory and anti-inflammatory activity. PMID- 8240468 TI - The epidemiology of intravenous vancomycin usage in a university hospital. A 10 year study. AB - OBJECTIVES: To examine the trends of intravenous vancomycin usage during a 10 year period, to classify the indications for which physicians prescribed the antibiotic, and to identify the independent predictors for empirical use of vancomycin. DESIGN: A descriptive epidemiological study, a cross-sectional study, and a case-control study were performed. SETTING: A 900-bed university-teaching hospital. MAIN OUTCOME MEASURES: The annual crude usage (grams) and incidence density (grams/1000 patient-days) of vancomycin were measured for 10 years (July 1981 to July 1991). In 109 randomly selected patient medical records, we evaluated the proportion of usage of vancomycin classified as prophylaxis, empirical therapy, or specifically directed therapy. Univariate and multivariate analyses were performed to identify determinants of empirical administration of vancomycin vs a penicillase-resistant penicillin to 64 case patients and 64 control patients. RESULTS: The rate of vancomycin usage increased 20-fold from 5.72 g/1000 patient-days in 1981 to 121.25 g/1000 patient-days in 1991. The use of vancomycin was significantly higher (P < .0001) in hematology-oncology areas compared with that in other hospital areas. The rates for each indication for vancomycin were 35.0% for prophylaxis 31.8% for empirical therapy, and 33.2% for therapy specifically directed by culture results. In a multivariate analysis, the presence of "plastic" medical devices was the best independent predictor for patients receiving vancomycin: intravenous lines (odds ratio [OR], 6.23; 95% confidence interval [CI], 2.28 to 17.06; P < .001), Hickman catheters (OR, 76.12; 95% CI, 15.06 to 384.73; P < .001), and other medical devices (OR, 10.50; 95% CI, 2.54 to 43.38; P = .001). CONCLUSIONS: Vancomycin use has increased linearly in the last decade primarily related to the presence of indwelling vascular devices in hematology-oncology patients. Use of vancomycin is equally divided among empirical therapy, prophylaxis, and specific therapy for a documented infection. PMID- 8240469 TI - Lack of association between sperm antibodies and recurrent spontaneous abortion. AB - The results of this investigation did not confirm the previous report by Witkin and Chaudhry (1). In contrast, they indicate that women with a history of recurrent SABs have a significantly lower incidence of circulating sperm antibodies. This finding is consistent with the considerable amount of evidence indicating that sperm antibodies can interfere with sperm function. PMID- 8240470 TI - Soft tissue edema in osteoid osteoma. AB - Four cases of osteoid osteomas are presented. An uncharacteristic magnetic resonance finding of soft tissue edema is reported. This observation should not be misinterpreted as indicating a more aggressive pathologic process and, thereby, exclude osteoid osteoma from the differential. PMID- 8240471 TI - Comparison of the reinforced laryngeal mask airway and tracheal intubation for adenotonsillectomy. AB - One-hundred and four patients were allocated randomly to receive anaesthesia for adenotonsillectomy via either a reinforced laryngeal mask airway or tracheal tube. Airway maintenance and protection were assessed during and after operation. The reinforced laryngeal mask did not interfere with surgical access; it resisted compression and protected the lower airway from contamination with blood. Four patients were withdrawn from the laryngeal mask airway group: two because difficulty with placement, and two because the laryngeal mask was obstructed distally when the Boyle Davis gag was opened fully. In children, recovery was less eventful in the laryngeal mask airway group, with less airway obstruction (P < 0.001) and better airway acceptance (P < 0.05). The reinforced laryngeal mask airway provided a clear, secure airway until recovery of protective reflexes. PMID- 8240472 TI - New program at Ontario hospital will screen "pre-embryos" for genetic disease. PMID- 8240473 TI - Managing substance use disorders in resident physicians. AB - Alcohol and other drug abuse is the leading cause of physician impairment. Prompt identification and early intervention by colleagues are the most effective ways of dealing with this problem in hospital residents. The author offers guidelines for recognizing the progressive signs of trouble inadvertently sent out by abusers, as well as effective ways in which colleagues can intervene. Also offered are steps in evaluation and treatment that can effectively address these illnesses and prevent compromising patient care. PMID- 8240474 TI - Recent trends in the identification and treatment of high blood cholesterol by physicians. Progress and missed opportunities. AB - OBJECTIVE: To investigate recent trends in the percentage and characteristics of patients being treated by a physician for high blood cholesterol (HBC) and to assess missed clinical opportunities to screen for HBC. DESIGN, SETTING, PARTICIPANTS: Telephone interviews of 154,735 adults in 37 states that participated in the Behavioral Risk Factor Surveillance System during 1988-1990 to assess trends in the percentage of patients treated for HBC by a physician. An opportunity was considered missed if a person did not report being screened for HBC despite seeing a physician for preventive care in the last 2 years. RESULTS: Between the first quarter of 1988 and the last quarter of 1990, the percentage of persons treated by a physician for HBC increased from 7.6% to 11.7% (P < .001). However, since an estimated 36% of US adults need treatment for HBC, fewer than one third of persons who need treatment are receiving it. Persons with two or more cardiac risk factors were more likely to be treated, while men, blacks, persons in lower socioeconomic groups, and persons between 20 and 34 years of age were less likely to be treated. Among the 126,571 persons who had seen a physician for preventive care within the last 2 years, missed opportunities to screen for HBC were most common among persons aged 20 through 34 years (59%) and among women who had seen obstetricians/gynecologists for preventive care (43%). CONCLUSIONS: Fewer than one third of persons who need treatment for HBC as estimated by data from the second National Health and Nutrition and Nutrition Examination Survey are receiving treatment. Better use of clinical opportunities to screen for HBC could substantially accelerate the progress in identifying persons, young adults in particular, who are likely to benefit from cholesterol reduction. PMID- 8240475 TI - Prevention of preterm birth: early detection and aggressive treatment with terbutaline. AB - We evaluated a program for prevention of preterm birth involving early detection and aggressive intervention with subcutaneous terbutaline pump therapy in a high risk, private patient population. Risk factor screening, frequent cervical examinations, and intensive patient education were used to detect preterm labor before it progressed to an advanced stage. Home terbutaline pump therapy was prescribed for patients with uterine contractions associated with progressive cervical change, after stabilization with IV magnesium sulfate. In this study of 51 patients, home terbutaline pump therapy was successful in 98% of the cases, prolonging pregnancy an average of 6.6 weeks. Mean gestational age at delivery was 37 +/- 1.4 weeks, and infant birth weight averaged 3 kg. Only 22% of infants required admission to the neonatal intensive care unit, with a mean length of stay of 7.25 days. Population factors in this compliant, well-educated patient group may have contributed to the positive outcomes achieved. PMID- 8240476 TI - Transverse myelitis following general anaesthesia. AB - A 47-year-old woman developed a complete transverse myelitis 2 weeks after an uncomplicated general anaesthetic for total abdominal hysterectomy and bilateral salpingo-oophorectomy. If spinal or epidural anaesthesia had been used, a causal relationship might have been assumed. This case emphasises the danger of attributing all cases of transverse myelitis which have a close temporal relationship to spinal or epidural anaesthesia, to the anaesthetic technique itself. PMID- 8240477 TI - Use of EMLA cream for skin anaesthesia prior to epidural insertion in labour. AB - The efficacy of the use of EMLA cream to provide skin anaesthesia prior to the insertion of 16 G Tuohy needles was assessed. Sixty women in labour were randomly allocated to receive either EMLA cream over the proposed epidural site for a minimum period of 5 min (mean 9.1 min), a skin bleb and subcutaneous injection of lignocaine 2% (1 ml), or a skin bleb with additional lignocaine 2% (2 ml) infiltrated into the supra- and interspinous ligaments using a 23 G needle. Once the epidural catheter was sited, using a standard technique, 10 cm visual analogue pain scales were completed independently by the patient, by a blinded midwife and by the anaesthetist siting the epidural. Patients' perception of pain was not significantly different in any of the three groups, although the mean pain score was least in the group receiving full infiltration. The anaesthetists consistently underestimated the patients' perception of pain. PMID- 8240478 TI - Overestimation of neonatal PO2 by collection of arterial blood gas values with the butterfly infusion set. AB - The butterfly, or scalp vein infusion set, is a widely used and convenient way to collect arterial blood gas samples in neonates. Analyzing arterial blood gas values by using this technique in a clinical situation, we obtained elevated values of PO2 in blood samples collected and measured from butterfly tubing in comparison with tuberculin syringe controls. Diffusion of gas through the polyvinylchloride tubing was suggested and proved by a study of pairs of butterfly and tuberculin syringes filled with venous blood from the blood bank. In conclusion, the butterfly gives falsely elevated PO2 values as a result of gas diffusion and is not a reliable technique for arterial blood gas sampling in neonates. PMID- 8240479 TI - A risk of using elastic adhesive bandage to secure the breathing circuit. PMID- 8240480 TI - The statistical versus clinical significance debate. AB - There is general agreement that tests of statistical significance do not provide information about the clinical significance or practical importance of research results. Yet the concept of clinical significance has received little discussion or debate in the nursing literature. The purpose of this paper is to compare and contrast statistical and clinical significance, to provide an overview of the issues surrounding their uses as described in the methodological literature from a variety of disciplines, and to discuss some implications of these concepts for nursing research. PMID- 8240481 TI - Recurrent obscure gastrointestinal bleeding. PMID- 8240482 TI - Lack of effect of ipriflavone on osteoclast motility and bone resorption in in vitro and ex vivo studies. AB - Using in vitro and ex vivo experimental procedures specifically designed to visualize pharmacological effects on parameters of bone resorption, studies were performed to elucidate whether ipriflavone's reported effect in osteoporosis is due to an effect on the motility and resorptive activity of osteoclasts, as has been shown to be the case with salmon calcitonin. Concentrations of ipriflavone used were higher by a factor of > 100 than peak blood levels measured in patients given standard therapeutic doses. Despite this, neither quantitative nor qualitative changes were observed in the motility of isolated rat osteoclasts or in their resorptive activity when incubated with bone slices. The conclusion is that ipriflavone does not possess antiosteoclastic and antiresorptive activity of the type documented for salmon calcitonin in the models employed and that further investigation of its mode of action is therefore necessary. PMID- 8240483 TI - Use of peer ratings to evaluate physician performance. AB - OBJECTIVE: To assess the feasibility and measurement characteristics of ratings completed by professional associates to evaluate the performance of practicing physicians. DESIGN: The clinical performance of physicians was evaluated using written questionnaires mailed to professional associates (physicians and nurses). Physician-associates were randomly selected from lists provided by both the subjects and medical supervisors, and detailed information was collected concerning the professional and social relationships between the associate and the subject. Responses were analyzed to determine factors that affect ratings and measurement characteristics of peer ratings. SETTING AND PARTICIPANTS: Physician subjects were selected from among practicing internists in New York, New Jersey, and Pennsylvania who received American Board of Internal Medicine certification 5 to 15 years previously. MAIN OUTCOME MEASURE: Physician performance as assessed by peers. RESULTS: Peer ratings are not biased substantially by the method of selection of the peers or the relationship between the rater and the subject. Factor analyses suggest a two-dimensional conceptualization of clinical skills: one factor represents cognitive and clinical management skills and the other factor represents humanistic qualities and management of psychosocial aspects of illness. Ratings from 11 peer physicians are needed to provide a reliable assessment in these two areas. CONCLUSIONS: These findings suggest that it is feasible to obtain assessments from professional associates of practicing physicians in areas such as clinical skills, humanistic qualities, and communication skills. Using a shorter version of the questionnaire used in this study, peer ratings provide a practical method to assess clinical performance in areas such as humanistic qualities and communication skills that are difficult to assess with other measures. PMID- 8240484 TI - Getting the story right. PMID- 8240485 TI - Collaborative tools. PMID- 8240486 TI - Chemical structures of the core regions of Campylobacter jejuni serotypes O:1, O:4, O:23, and O:36 lipopolysaccharides. AB - Complete structures, including the location of N-acetylneuraminic acid (Neu5Ac) residues, were assigned for the core regions of Campylobacter jejuni serotypes O:1, O:4, and O:23 and O:36 lipopolysaccharides (LPS). In continuation of earlier studies, structure determinations of liberated oligosaccharides and, where necessary, of intact LPS, were by 1H-NMR spectroscopy, Smith degradation, chromium trioxide and enzymic degradations, in conjunction with methylation studies supported by fast-atom-bombardment mass spectrometry and linkage analyses by gas chromatography/mass spectrometry. It was concluded on the basis of the following structures, in which each was linked 1-->5 to a terminal 3-deoxy-D manno-octulosonic acid residue, that the core regions with qualitatively similar sugar compositions showed serotypic differences in one or more of their sequences, linkage types, and anomeric configurations: [formula: see text] [corrected]. The outer regions of each structure carry Neu5Ac residues linked 2- >3 to available beta-D-Galp residues and show striking similarities with various glycosphingolipids of the ganglioside family. However, Neu5Ac epitopes are not apparently involved in determining serospecificity. PMID- 8240487 TI - Specialist medical training and the Calman report. PMID- 8240488 TI - Secondary centres should provide neonatal intensive care. PMID- 8240489 TI - The outcome of Charnley total hip arthroplasty with cement after a minimum twenty year follow-up. The results of one surgeon. AB - We evaluated the results of 330 total hip arthroplasties that were performed with use of the Charnley prosthesis and cement in 262 patients by the senior one of us between July 1970 and April 1972. All hips had been thoroughly assessed preoperatively to document the patient's functional level. All patients had been disabled because of pain in the hip or a fracture of the hip, and 212 patients (81 per cent) had used walking aids. At a minimum of twenty years after the index operation, eighty-three patients (ninety-eight hips) were still living, 174 patients (224 hips) had died, and five patients (eight hips) had been lost to follow-up. The outcome of the arthroplasty was determined for all except the five latter patients. Thus, the outcome of 322 (98 per cent) of the 330 arthroplasties was known at the latest follow-up evaluation. Radiographs were available for sixty-three of the eighty-three patients (seventy-six [78 per cent] of the ninety eight hips) who were alive for the entire follow-up period. Of the ninety-eight hips in the living patients, eighty-three (85 per cent) caused no pain, fourteen (14 per cent) caused mild pain, and one (1 per cent) caused moderate pain. Fifty two hips (53 per cent) were in patients who did not use walking aids, and only seven (7 per cent) were in patients who used support for walking because of the hip. At the minimum twenty-year follow-up, thirty-two (10 per cent) of the 322 hips that had been followed had been revised: eight (2 per cent), because of loosening with infection; twenty-one (7 per cent), because of aseptic loosening; and three (1 per cent), because of dislocation. Of the ninety-eight hips of the patients who were still alive, fifteen (15 per cent) had been revised: three (3 per cent), because of loosening with infection; eleven (11 per cent), because of aseptic loosening; and one (1 per cent), because of dislocation. The rate of revision due to aseptic loosening of the acetabular component in all 322 hips was 6 per cent (eighteen hips), while in the ninety-eight hips of the patients who were alive at least twenty years after the arthroplasty, it was 10 per cent (ten hips). The rate of revision because of aseptic loosening of the femoral component in all 322 hips was 2 per cent (eight hips), while in the ninety-eight hips of the living patients, it was 3 per cent (three hips).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8240490 TI - Interposed abdominal compression--cardiopulmonary resuscitation. PMID- 8240491 TI - Induced hypothermia for sepsis? PMID- 8240492 TI - Diagnostic difficulties in community-acquired pneumonia. PMID- 8240493 TI - J. vaquero and R. Arriaza: the patella thinning osteotomy. PMID- 8240494 TI - Acute renal failure in severe exertional rhabdomyolysis. PMID- 8240495 TI - Falciparum malaria and jaundice. PMID- 8240496 TI - Cobalamin deficiency in the elderly. PMID- 8240497 TI - The immediate effect of manipulation vs. mobilization on pain and range of motion in the cervical spine: a randomized controlled trial. PMID- 8240498 TI - Regression toward the mean in 2 x 2 crossover designs with baseline measurements by Grender, J. M., Johnson, W. D. and Elston, R. C., Statistics in Medicine, 11, 727-741 (1992) PMID- 8240499 TI - Is access to health care a right? PMID- 8240500 TI - Pharmacists number more than 190,000 in United States. PMID- 8240501 TI - Blind biopsy of the contralateral breast at the time of the initial treatment of breast carcinoma is not an efficient method of cancer detection. PMID- 8240502 TI - Cautery for lower lid entropion. PMID- 8240503 TI - Open access gastroscopy. Open access means overinvestigation. PMID- 8240504 TI - Control of communicable disease. Consultants need managerial and financial authority. PMID- 8240505 TI - Putting patients first. PMID- 8240506 TI - Neonatal intensive care. International network provides feedback. PMID- 8240507 TI - The Hawksley random zero sphygmomanometer. Repeat experiment exonerates instrument. PMID- 8240508 TI - Medical angiologists. Britain needs them. PMID- 8240509 TI - Teaching communication skills. Funding required for teaching programmes. PMID- 8240510 TI - Clozapine in the management of schizophrenia. Clozapine has unique pharmacological profile. PMID- 8240511 TI - Home treatment for acute psychiatric disorder. A marvellous advance. PMID- 8240512 TI - Re: Effect of vaginal ultrasound probe on lower urinary tract function. PMID- 8240513 TI - [Poliomyelitis in The Netherlands, 1979-1991: immunity and exposure]. AB - An overview is presented of serological and virological studies on poliovirus immunization and circulation in the Netherlands, performed between 1979 and 1991. In this period, only three patients with poliomyelitis were notified. All had acquired the infection abroad. The vaccinations in the national immunization programme, using inactivated poliovirus vaccine, build a strong immunity. This can also be seen in age-stratified serological profiles of the Dutch population. In these surveys, persons from the time at which vaccination was offered have neutralizing antibodies. Older persons, especially those born between 1930 and 1945, sometimes lack antibodies. However, 85-90% of them show a rapid booster response upon vaccination, demonstrating immunological memory. Hence, they will be protected against poliomyelitis upon contact with wild poliovirus. Virological data show a regular import of poliovirus, especially in adoptive children tested on entry into the Netherlands, coming from developing countries. Nearly all other virus isolates in Dutchmen were related to import from such countries. None of the imported patients or other persons in whom poliovirus was detected spread the virus over the country. It demonstrates that as a rule the herd immunity of the well-vaccinated Dutch population is good. Exceptions occur, however, as demonstrated by the epidemics in 1978 and 1992. Large socio-geographic clusters of susceptible people who refuse vaccinations are not sufficiently protected. PMID- 8240514 TI - Article distracts nurses... PMID- 8240515 TI - Parvovirus and neonatal hydrops. PMID- 8240516 TI - Determining carcinogenicity. PMID- 8240517 TI - Acute monocytic leukemia with chloroacetate esterase positivity: FAB M4 or M5? PMID- 8240518 TI - Stylet for reinforced laryngeal mask airway. PMID- 8240519 TI - Pain and the paramedian approach to epidural catherisation. PMID- 8240520 TI - Vomiting after alfentanil anesthesia. PMID- 8240521 TI - Total anomalous pulmonary venous connection. PMID- 8240522 TI - Prognostic value of the S-phase fraction of breast cancer. PMID- 8240523 TI - Opioid/autonomic links in disease. PMID- 8240524 TI - Reforming our health care system. PMID- 8240525 TI - Spinal cord ataxia. PMID- 8240526 TI - Hereditary cerebral hemorrhage with amyloidosis-Dutch type. PMID- 8240527 TI - Antithyroid antibodies in families with Alport's syndrome. PMID- 8240528 TI - Clinical ethics in family practice. PMID- 8240529 TI - Surgical back-up in percutaneous transluminal coronary angioplasty. PMID- 8240530 TI - Hold the talc, pass the cornstarch. PMID- 8240531 TI - Clinical vs nuclear laboratory diagnosis of congestive heart failure. PMID- 8240533 TI - Experiences with GCP in the pharmaceutical industry [corrected]. AB - The Declaration of Helsinki and the "new" Germany Drug Law of 1976 may be regarded as the first and all important step in the evolution of Good Clinical Practices (GCP) in Germany. "Regulations on the orderly conduct of clinical studies" (published in 1987), reflecting most elements of the GCPs, caused the German industry to make provisions for more successful drug approvals in the future. Pharmaceutical companies have improved their documentation systems with regard to study protocols and reports, increased their monitoring capacity, and began setting up SOPs. In preparation for the issuing of the E.C. GCP guidelines companies established Clinical Quality Assurance (CQA) units responsible for various CQA tasks such as training of monitors and investigators, auditing of protocols, reports, systems, and data. Typical problems connected with the implementation of GCP in Germany appear to be contradictory opinions of the various ethics committees, reluctancy on the part of investigators to assume additional bureaucratic workload, difficulties on the part of the sponsor resulting from the high financial and manpower requirements, as well as the relatively unfavorable political and public opinion of clinical research. The benefits achieved by the introduction of GCP are already visible, i.e., increased quality of study protocol, conduct and report of studies, reduced number of studies, transparency at all stages of clinical research and increased ethical awareness. PMID- 8240532 TI - N-acetylglucosaminyltransferase III and V messenger RNA levels in LEC rats during hepatocarcinogenesis. AB - The LEC (Long-Evans with a cinnamon-like color) rat is a mutant of the Long-Evans strain which develops hereditary hepatitis and hepatoma with age. Activities and mRNA levels of N-acetylglucosaminyltransferase III and V (GnT-III and GnT-V, respectively) were determined during hepatocarcinogenesis in this rat using a LEA (Long-Evans with an agouti color) rat as a control. GnT-III activity in LEC rat liver increased after 30 weeks of age, at the stage of chronic hepatitis, to about 2.5-11.5 times the level in LEC rats aged 1-9 weeks. GnT-V activity in the LEC rat liver increased after 20 weeks of age, at the stage of acute hepatitis, to about 1.5-2.5 times the level in LEC rats of 1-9 weeks of age and then remained elevated. Both enzymes showed more dramatic increases in males than in females. The mRNA levels of the enzymes increased in proportion with the enzyme activities. Furthermore, GnT-III and GnT-V mRNAs were highly expressed in both cancer lesion and adjacent tissues. In one case of hepatoma with lymph node metastasis, GnT-III and GnT-V mRNA expression was much higher in the metastatic lesion than in the original cancer. GnT-III and GnT-V levels in the original cancer lesions were similar to those in the cancer lesions of the other LEC rats. These results indicated that expression of GnT-III and GnT-V was induced by chronic liver damage and hepatocarcinogenic changes in the LEC rats. PMID- 8240534 TI - [Double-blind study versus excipient of 0.75% metronidazole gel in the treatment of rosacea]. AB - INTRODUCTION: The proven value of tetracyclines and metronidazole administered orally in the treatment of the chronic and recurrent disease that is rosacea is tempered by the important undesirable effects observed in long-term therapy. The purpose of this study was to test the effectiveness of an 0.75 p. 100 metronidazole gel in the treatment of rosacea. PATIENTS AND METHODS: The study involved two groups of patients: one received the metronidazole gel and the other the vehicle of the gel used as placebo. The multicentre randomized trial was conducted in the double-blind fashion by 18 private dermatologists working in the Paris region. Fifty one patients who, since more than 3 months, had been presenting with rosacea, defined as at least 4 papulopustules associated with erythema and/or telangiectasia, entered the trial. Topical treatments and systemic treatments which had shown some activity against rosacea had been interrupted for 15 days or 2 months respectively. The product (or the placebo) was applied a.m. and p.m. on the whole dry face. The patients were seen on days 0, 21 and 42. The evaluation was purely clinical, and the principal criterion of judgement was a change in the number of papulopustules between days 0 and 42. RESULTS: The metronidazole gel reduced the number of papulopustules between day 0 and day 42, and this reduction was significantly greater than that observed with the excipient alone. The active product began to be effective during the third week and remained so during the next three weeks. Both the metronidazole gel and its excipient seemed to be poorly tolerated, with frequent complaints of dry skin, but in 5 women of the metronidazole group this dryness was alleviated by application of moisturizers. CONCLUSION: This study has demonstrated that the 0.75 p. 100 metronidazole gel is effective in the treatment of the papulopustular component of rosacea. PMID- 8240535 TI - Political correctness or primum non nocere? PMID- 8240536 TI - Terbutaline pump treatment of premature labor. PMID- 8240537 TI - Absence of excess chromosome breakage in a patient with VACTERL-hydrocephalus. PMID- 8240538 TI - Site of pharyngeal narrowing predicts outcome of surgery for OSA. PMID- 8240539 TI - Vasovagal syncopy related to peritoneal dialysate infusion. PMID- 8240540 TI - Management of lethal catatonia with dantrolene sodium. PMID- 8240541 TI - Comments on "training Japanese listeners to identify English /r/ and /l/: a first report" [J.S. Logan, S.E. Lively, and D.B. Pisoni, J. Acoust. Soc. Am. 89, 874 886 (1991)]. PMID- 8240542 TI - Primary abdominal actinomycosis. PMID- 8240543 TI - Greenwald's law of lupus: the Sontheimer amendment. PMID- 8240544 TI - Futility and avoidance: medical professionals in the treatment of obesity. PMID- 8240545 TI - OBRA fee reduction and physician behavior. PMID- 8240546 TI - [Maintaining patency of a coronary bypass: why doing it the hard way?]. PMID- 8240547 TI - Conversion of gastrostomy tube to gastrojejunostomy tube. PMID- 8240548 TI - Angiotensin II-induced increases in platelet intracellular free calcium in hypertensive and normotensive pregnancy. PMID- 8240549 TI - [Phlebitis of the upper limbs on pacemaker electrodes. 4 cases and review of the literature]. AB - We report four cases of deep venous thrombosis of the upper extremity, which occurred in two women and two men (mean age 79 years) in whom a pacemaker electrode had been inserted 4 years on averages previously. In three of these four cases phlebitis developed after immobilization of the limb containing the electrode. Deep venous thrombosis of the upper extremity is rare, but 28% of catheterizations are responsible for phlebitis. One to 3% of patients fitted with a pacemaker have symptomatic phlebitis, but these figures rise to 28-65% when phlebography is systematically performed in subjects wearing a pacemaker. The clinical signs are the same as those of the classical forms, and the diagnosis is made by doppler-ultrasonography and by phlebography which informs on the collateral circulation. Cure is obtained with efficient anticoagulant therapy. These cases prompt us to prescribe a preventive subcutaneous heparin therapy in those pacemaker-fitted subjects whose arm is immobilized. The heparin dosage remains to be determined precisely. PMID- 8240550 TI - Gastroduodenal ulceration associated with flunixin meglumine administration in three dogs . AB - In 3 clinically ill dogs, signs of gastroduodenal ulceration were first noticed within 7 days of beginning flunixin meglumine administration and included pyrexia, anorexia, weight loss, vomiting, melena, pain on abdominal palpation, and abdominal distention. One dog was euthanatized and 2 dogs recovered after surgical repair of the perforated ulcers and treatment of peritonitis. Prolonged administration of flunixin meglumine should be avoided, especially in debilitated dogs or when concurrently administering other nonsteroidal anti-inflammatory drugs or corticosteroids. PMID- 8240551 TI - The 1.7 A refined X-ray structure of the periplasmic glucose/galactose receptor from Salmonella typhimurium. AB - The X-ray structure of the periplasmic glucose/galactose receptor (binding protein) of Salmonella typhimurium (GBP-S) has been refined at 1.7 A resolution with an R-factor of 19.0%. The model contains all 309 residues of the amino acid sequence, 153 water molecules, a calcium ion and beta-D-galactose. The protein consists of two very similar structural domains, each of which is composed a core of parallel beta-sheet flanked on both sides by alpha-helices. Three short stretches of amino acid chain (from symmetrically related portions of the structure) link the domains, and presumably act as a hinge to allow their relative movement in functionally important conformational changes. Galactose is bound between the domains, interacting with a number of side-chains from the loops lining the binding cleft. A combination of hydrogen bonding, hydrophobic and steric effects give rise to tight binding (dissociation constant 0.2 microM) and high specificity. Of nine hydrogen bonding groups, three are aspartate, three asparagine, one histidine (unprotonated), one arginine and one water, contributing 13 hydrogen bonds in total. Additional residues pack against (primarily) non-polar faces of the sugar molecule. The precise arrangement of the hydrogen bonding and hydrophobic residues results in an enclosed binding site with a shape that is a composite of those of the allowed sugar molecules. It is presumed that ligands bind to a more open form of the receptor that then closes by rotation in the hinge. Comparison with the GBP-S structure solved earlier in complex with glucose shows no significant changes, even for the aspartate residue most directly involved with the different sugars. Comparison with the galactose/glucose receptor of Escherichia coli indicates that these two proteins are very similar in overall structure, with the main difference being a 2 to 3 degrees rotation in the hinge. This difference appears to be the result of different crystal packing for the two proteins; it is likely that both conformations are normally found in solution. PMID- 8240552 TI - [Management of HIV seropositive patients. Participation of general practitioners in the Puy-de Dome in 1992]. PMID- 8240553 TI - Differentiating acute appendicitis from pelvic inflammatory disease in women of childbearing age. AB - A retrospective study was performed to evaluate the usefulness of various historical, clinical, and laboratory findings in differentiating acute appendicitis from pelvic inflammatory disease (PID) in women of childbearing age. The records of all female patients presenting to the emergency department with abdominal pain who were found to have histologically proven appendicitis (n = 80) or PID confirmed on endocervical culture (n = 71) were reviewed. Clinically useful indicators favoring appendicitis included the presence of anorexia and the onset of pain later than day 14 of the menstrual cycle. Indicators favoring PID included a history of vaginal discharge, urinary symptoms, prior PID, tenderness outside the right lower quadrant, cervical motion tenderness, vaginal discharge on pelvic examination, and positive urinalysis. Despite these indicators, differentiating acute appendicitis from PID remains difficult. PMID- 8240554 TI - Ketorolac as a rapid and effective treatment of migraine headache: evaluations by patients. AB - The objective of this study was to determine the speed and efficiency of ketorolac in reducing the symptoms of migraine headache. Twenty-three patients who presented in the emergency department during the period between April and July 1992 with a previous diagnosis of migraine headache were considered for the study. Patients subjectively evaluated parameters of their migraine headaches (eg, pain and nausea) with a numerical scale and were asked to re-evaluate these same parameters at 30, 60, and 360 minutes after a single injection of Ketorolac. Seventeen (74%) patients reported a decrease in headache symptoms that was significant (P < .005) after 1 hour. Relief lasted at least 6 hours after injection. PMID- 8240555 TI - Expediting the early hospital care of the adult patient with nontraumatic chest pain: impact of a modified ED triage protocol. AB - A prospective study that compared a traditional emergency department (ED) triage protocol with an expedited protocol was conducted to determine if minimizing the subjectivity of nursing triage would result in more efficient management of adult patients presenting with nontraumatic chest pain. The traditional protocol triaged 382 patients into 1 of 5 categories of acuity. The expedited study group (418 patients) were triaged as usual but subsequently were treated as if they were triage category 1 or 2 (medical evaluation within 15 minutes of arrival). Traditional triage led to 40% of acute myocardial infarction (AMI) patients being triaged into inappropriately low-acuity categories. The expedited protocol resulted in significant improvement in the following intervals: ED arrival to triage, triage to cubicle, ED arrival to cubicle, ED arrival to electrocardiogram (ECG) ordered, ED arrival to ECG available, ED arrival to physician evaluation, and ED arrival to decision to thrombolyse. Study patients with non-AMI cardiac chest pain and AMI cardiac chest pain were evaluated by a physician an average of 12 minutes and 8 minutes after ED arrival, respectively. Delays in interdepartmental processes, such as ECG-technician responsiveness, thrombolysis protocol fulfillment and thrombolytic agent delivery, negated benefits derived from improvements in internal processes. Effective coordination of the numerous processes involved in the initial ED management of adult patients with nontraumatic chest pain is required to make thrombolytic therapy for AMI within 30 minutes of patient arrival a routinely achievable goal. PMID- 8240556 TI - Activated charcoal surface area and its role in multiple-dose charcoal therapy. AB - The study hypothesis was to prove that increasing the surface area (SA) of activated charcoal (AC) will enhance theophylline elimination in multiple-dose AC therapy. Five healthy, nonsmoking, nonmedicated, volunteer men ranging from the ages of 18 to 24 years old were entered onto the study. A prospective, randomized, crossover study was conducted with each subject serving as their own control. Subjects fasted overnight before receiving 8 mg/kg of intravenous theophylline at the beginning of a control phase and two study phases. No AC was administered in the control phase. Two experimental phases compared Actidose Aqua AC (1,500 m2/g; Paddock Laboratories, Inc, Minneapolis, MN) with Norit A Supra AC (2,000 m2/g; Norit NV, Amersfoort, the Netherlands). In each phase, 50 g of the AC was administered orally after the conclusion of the theophylline infusion. Subsequent 50-g doses of the AC preparation were administered at 4 and 8 hours after the initial dose of AC. Serial blood samples for theophylline level determinations were obtained at 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 10.0, and 12.0 hours after the completion of the theophylline infusion. For each of the three data sets, the absolute area under the absorption curve (A-AUC) was calculated to infinity using the trapezoidal rule. Additionally, the relative area under the absorption curve (R-AUC) was determined for each data set. Data were analyzed using repeated measures of analysis of variance (ANOVA) and Tukey's test. The alpha error was set at 0.05.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240557 TI - Reimbursement patterns in a hospital-based fixed-wing aeromedical service. AB - Reimbursement, expressed as a percentage of total charges recovered, was examined for inpatients transported by a university hospital-based, dedicated, fixed-wing aeromedical service between July 1, 1988 and June 30, 1990. A total of 410 patients were transported; account information was available for 404 patients (98%). Patients transported from in-state institutions (n = 174) had a hospital reimbursement rate of 53.3%, whereas the flight program recovered 46.1% of transportation charges. Patients transported from out-of-state institutions (n = 150) had a hospital reimbursement rate of 51.3%, whereas the flight program recovered 69.3% of charges. More patients referred from in-state sources were covered by Medicaid than from out-of-state sources (31% vs 11%), and less were covered by Medicare (17% vs 30%). Reimbursement for hospital charges was low for patients covered by Medicaid (44% for in-state and 16% for out-of-state), and Medicaid reimbursed no flight charges for either in-state or out-of-state patients. The flight program recovered 86.7% of charges for "triangle" flights, which transported patients between two hospitals other than the sponsoring institution (n = 80). The overall hospital reimbursement rate for inpatients was 70% during the study period. The flight program recovered 53.7% of its operating costs from payment of charges for transport services. Aeromedically transported patients may represent a financial "high-risk" group of patients for the sponsoring institution. PMID- 8240558 TI - A radiographic comparison of short-arm cast and plaster and fiberglass wrist splints. AB - Prepackaged plaster and fiberglass splints are used in many emergency departments. This study evaluated the effectiveness of short-arm cast (SAC), volar fiberglass wrist splint (FWS), and volar plaster wrist splint (PWS) in limiting wrist range of motion. Ten healthy male volunteers between the ages of 18 and 35 years were included. Each wrist on each volunteer was immobilized with SAC, FWS, and PWS. Wrist radiographs were taken with each appliance and angular motion measured by two radiologists. PWS performed better than FWS in flexion, extension, radial deviation, and ulnar deviation (all P < .05). PWS was not statistically different than SAC in limiting flexion, extension, or radial deviation, although SAC performed better in ulnar deviation (P < .05). PWS limits wrist motion more effectively than FWS and performs in a similar manner to SAC in flexion, extension, and radial deviation. PMID- 8240559 TI - Standards and limits: emergency physicians' attitude toward prehospital resuscitation. AB - The attitudes of emergency physicians toward using medical techniques in out-of hospital medical cardiac arrest scenarios were assessed. The physicians' willingness to further limit the use of these techniques through prehospital Do Not Attempt Resuscitation (DNAR) protocols and their personal use of advance directives were also assessed. Questionnaires were distributed to the 1990 Council of the American College of Emergency Physicians, San Francisco, CA, to elicit demographic and clinical information. Of the 172 ACEP Council members (emergency physicians) asked to participate, 136 (79%) returned a questionnaire. These physicians represented all of the 50 United States, Puerto Rico, and two Canadian provinces. In two scenarios, most respondents recognized the uselessness of continued resuscitation for themselves, their spouse, and their parent. However, in these same situations approximately half of them would continue treating their child. Although nearly two thirds of the respondents did not have a prehospital DNAR policy in their emergency medicine system (EMS), 95% felt that they needed such a protocol. Only one-third of respondents had any type of advance directive for themselves. Experienced emergency physicians recognize that there are limits to the application of medical techniques, but are less willing to stop pediatric resuscitations, than they are to stop adult resuscitations. Emergency physicians agree on the need for prehospital DNAR protocols, but few have completed their own advance directives. PMID- 8240560 TI - Differential front and back seat safety belt use by prehospital care providers. AB - The object of the study was to assess the habits and attitudes of prehospital care personnel regarding safety belt use in the front and rear ambulance compartments. Therefore, a cross-sectional descriptive survey was administered at emergency medical service conferences and through provider agencies throughout the United States and Canada. Approximately 900 public, private, and volunteer prehospital care providers participated. Demographic information, traffic collision history, percent of time safety belts were used, belief in safety belt use, and reasons for nonuse in the rear compartment of the ambulance were measured. The results showed that safety belt use was highest in the front seat during emergency runs (median, 100%) and rarest in the back compartment during emergency runs (median, 0%). Respondents cited the following reasons for non-use in the rear compartment: inhibited patient care (67.9%), restricted movement (34.7%) inconvenience (15.1%), or lack of efficacy (5.3%). Prehospital care personnel typically wear safety belts when in the front seat, but not while in the rear compartment of the ambulance. More intensive efforts at educating prehospital care providers about the importance of safety restraints in the rear compartment, enumerating patient care activities that can be performed while wearing a safety belt, and design of a functional restraint system for the rear compartment may increase ambulance safety. PMID- 8240561 TI - Case report of factor VII deficiency. AB - The case of a 57-year-old woman with no personal or family history of coagulopathy or blood dyscrasia who was found to be factor VII deficient by routine laboratory testing is reported. The patient was also found to have type 2 diabetes mellitus and adenocarcinoma of the uterus in the course of her hospitalization. PMID- 8240562 TI - Complete heart block in a child with varicella. AB - A case of varicella myocarditis in a previously healthy 6-year-old child was reviewed. The patient presented with third-degree heart block and shock as the sole manifestation of her cardiac involvement. Bradyarrhythmias required temporary transvenous pacing. Intravenous acyclovir was used. The patient recovered without permanent sequelae. The natural history, clinical presentation, and treatment of varicella myocarditis are reviewed. PMID- 8240563 TI - Contact lens abrasions and the nonophthalmologist. AB - Corneal abrasions associated with contact lens wear are commonly evaluated and treated in acute care clinics and emergency departments by nonophthalmologists. The risk of progression to suppurative keratitis in this setting requires management distinct from that of other mechanical (eg, fingernail scratch) corneal abrasions. The antibiotic chosen should reflect the need for prophylaxis against Pseudomonas. Conditions favoring bacterial growth, specifically occlusive patching and/or use of steroid containing compounds, should be avoided, and 24 hour follow-up examination is recommended. PMID- 8240564 TI - Ondansetron quells drug-resistant emesis in theophylline poisoning. PMID- 8240565 TI - Traumatic rupture of the stomach secondary to Heimlich maneuver. AB - The case of a 93-year-old man who received a Heimlich maneuver while choking is reported. After the procedure, the patient presented with abdominal pain and ultimately was found to have developed a gastric rupture. He was hospitalized for 66 days. Review of the literature showed that only four gastric perforations related to the Heimlich maneuver have been documented. Other complications have occurred. It is reasonable to perform the procedure as an alternative to asphyxiation, but emergency physicians must be aware of the fact that life threatening complications may ensue. PMID- 8240566 TI - Inadvertent administration of rtPA to a patient with type 1 aortic dissection and subsequent cardiac tamponade. AB - Administration of thrombolytic agents to patients with misdiagnosis of acute myocardial infarction can result in serious side effects. A case of aortic dissection that was misdiagnosed as acute myocardial infarction and received rtPA is reported. Within 1 hour of rtPA infusion, the patient developed cardiac tamponade. Type 1 aortic dissection was diagnosed by aortic angiogram. The patient underwent emergency resection of the dissection and evacuation of the pericardial and anterior mediastinal hematoma. Although he required massive transfusion of blood products intraoperatively, he is doing well 22 months after his surgery. PMID- 8240567 TI - Recombinant erythropoietin overdose. AB - Deliberate self-administration of recombinant human erythropoietin (rHuEpo) in a patient without anemia has never been documented. The case of a 62-year-old man who worked in an allied health care field and surreptitiously injected the drug, causing his hematocrit to increase to a dangerously high level is presented. Resultant complications of the misuse of erythropoietin in this patient included worsening hypertension, exacerbation of chronic lung disease and development of new onset angina. Medical management consisted of endotracheal intubation with mechanical ventilation, intravenous hydration, and serial phlebotomy. The unusual possibility of erythropoietin abuse must be added to the differential diagnosis with a patient with unexplained polycythemia. This case highlights the potential abuse of biological growth factors that may mask medical conditions. PMID- 8240568 TI - Gas gangrene from subcutaneous insulin administration. AB - A case of gas gangrene that caused intractable shoulder pain refractory to narcotics in an immunocompromised host is presented. Gas gangrene has been associated with severe trauma involving penetrating wounds, compound fractures, extensive soft-tissue injury, intramuscular injection of epinephrine, and interruption of arterial blood supply. This case describes an elderly insulin dependent diabetic woman who developed gas gangrene in her arm and leg at the site of her subcutaneous insulin injections. The responsible organism was Clostridium septicum. Emergency medicine physicians must consider gas gangrene Clostridium infection in immunocompromised individuals without evidence of trauma who present with localized and intractable pain. PMID- 8240569 TI - Psychogenic convulsive seizures. PMID- 8240570 TI - Hemorrhagic varicella: a case report and review of the complications of varicella in children. AB - The case of a previously healthy child who developed progressive systemic varicella with purpura is reported. The clinical course of this patient is outlined, and the range of potential complications of chickenpox in children is reviewed. Familiarity with the usual uncomplicated natural history of primary varicella infection should alert the clinician to signs and symptoms that signal significant systemic involvement. PMID- 8240571 TI - A spectrophotometric method for estimating methemoglobin concentration in the presence of cyanide. PMID- 8240572 TI - ED management of acute congestive heart failure in renal dialysis patients. AB - This is a descriptive report of the management techniques used effectively in the emergency department (ED) treatment of acute congestive heart failure (CHF) in renal dialysis patients. Study design included a prospective case series of consecutive renal dialysis patients who presented to the ED of a regional dialysis center in acute CHF. Clinical presentation, ED management, and outcome were recorded. Forty-six patients (38 hemodialysis and 8 peritoneal dialysis) were included in this study. Presentation classifications for these patients were minimal distress (13 patients), moderate distress (16 patients), and severe distress (17 patients). In addition to supplemental oxygen, treatment focused on pharmacological preload and afterload reduction. Patients received sublingual nitroglycerin (NTG) (30 patients), transdermal NTG (36 patients), captopril sublingual (10 patients) nifedipine oral (nine patients), nitroprusside (four patients), morphine sulfate (one patient), NTG infusion (one patient), and clonidine (one patient). There were no deaths in the study group, and 32 of the patients were able to be dialyzed and discharged, including seven patients in the severe group. Six patients required intubation, one of whom was extubated and discharged from the ED after dialysis. Intravascular access was obtained in 29 patients but was used in only six. All patients on nitroprusside drips were weaned during the course of their dialysis. Effective ED management of acute CHF in renal dialysis patients can be accomplished through preload reduction with nitrates and afterload reduction with captopril, nifedipine, and, in severe cases, nitroprusside. PMID- 8240573 TI - The efficacy and acceptability of using a jet injector in performing digital blocks. AB - This prospective, nonblinded study, comparing jet injection with needle-syringe injection of lidocaine in performing digital blocks, tested the hypothesis that jet injection can be used effectively as a less painful way to perform digital blocks. Twenty-four adult patients with injuries of the middle or distal phalanges of the fingers received digital blocks using a jet injector on one side of the finger and a needle-syringe on the other side; pain was assessed at 0, 1, 3, 6, 12, and 24 hours using visual analog scales. Differences in pain scores for the two procedures (jet injector vs needle) were tested at each time period using nonparametric statistical procedures for paired or matched data (paired Wilcoxon). At the time of injection (time 0) and at 12 hours, the jet had a significantly lower pain score than the needle. Comparisons made at the other points were statistically nonsignificant at .05. The anesthesia achieved using the jet injector was considered adequate in 23 of 24 patients. We conclude that the jet injector can be used effectively in performing digital blocks and is less painful than standard needle-syringe methods. PMID- 8240574 TI - Interhospital transfer of cardiac patients by air. PMID- 8240575 TI - Intravenous urapidil versus sublingual nifedipine in the treatment of hypertensive urgencies. AB - In a 6-month prospective study, the efficacy and safety of urapidil and nifedipine in an outpatient population with hypertensive urgencies (systolic blood pressure > 200 mm Hg; diastolic blood pressure > 110 mm Hg) was investigated. Response to treatment was defined as a stable reduction of systolic blood pressure below 180 mm Hg and diastolic blood pressure below 100 mm Hg 15 minutes after application of a single dose of either 25 mg urapidil intravenously (N = 26) or 10 mg nifedipine sublingually (N = 27). If the blood pressure was still elevated, a second dose of 10 mg nifedipine or 12.5 mg urapidil was given, and blood pressure response was evaluated 15 minutes after application of the second dose according to the aforementioned criterias. After the first application of nifedipine, 19 (70%) responders have been observed. Eight patients needed an additional 10 mg of nifedipine. In four of these patients, no reduction of blood pressure was observed after a second dose of nifedipine. In contrast, 24 (92%) patients responded well to the first application of 25 mg of urapidil. Two patients required a second dose of 12.5 mg of urapidil, but no nonresponder to urapidil was observed. No severe side-effects were noted in both groups. Intravenous urapidil is a highly effective drug in the treatment of hypertensive urgencies and is more effective than sublingual nifedipine, because the number of patients treated successfully was significantly higher. PMID- 8240576 TI - Critical care medicine: an annotated bibliography of the recent literature. PMID- 8240577 TI - Undiagnosed hypothyroidism: a risk factor for digoxin toxicity. PMID- 8240578 TI - Comparison of ED visually and meter-read Chemstrip bG test strips. PMID- 8240579 TI - Acute leukemia presenting coexistent with endocarditis. PMID- 8240580 TI - The noise hazard of treating children in the ED. PMID- 8240581 TI - High incidence of psychosis in cocaine intoxication and preventing violence in the ED. PMID- 8240582 TI - Subcutaneous gasoline injection. PMID- 8240583 TI - Pneumothorax secondary to a dog bite. PMID- 8240584 TI - Gun control: a simple answer to the wrong question. PMID- 8240585 TI - Cocaine-associated myocardial infarction secondary to a contaminant. PMID- 8240586 TI - Paroxetine overdose. PMID- 8240587 TI - The Quality Assurance Committee. PMID- 8240588 TI - Hypoxanthine in fetal umbilical venous blood and amniotic fluid from pregnancies complicated by rhesus isoimmunization. AB - Concentrations of hypoxanthine (HX) was determined in umbilical venous blood and amniotic fluid obtained at 74 instances in 36 rhesus immunized patients before the onset of labor. HX concentrations were related to gestational age, concentrations of hemoglobin and lactate, pH, and partial oxygen pressure in umbilical venous blood. Multiple regression analysis revealed hemoglobin concentration to be the only variable that had any explanatory power to HX in amniotic fluid. No one of the studied variables gave any significant contribution to a regression model to explain HX in umbilical venous blood. We conclude that HX levels in umbilical venous blood and in amniotic fluid from rhesus immunized patients were not associated with fetal blood gases before the onset of labor. PMID- 8240589 TI - Emergency cervical cerclage: a retrospective review of 51 cases. AB - Fifty-one patients underwent emergency cervical cerclage using Shirodkar or McDonald suture for incompetent cervix diagnosed during pregnancy. Fifteen patients had only sonographic evidence of cervical incompetence (group I). Eighteen patients had cervical dilation of less than 3 cm (group II), and 18 patients had cervical dilation of 3 cm or more (group III). Perinatal mortality was 0%, 50%, and 55.6%, respectively. The median length of time gained with the cerclage was 16 weeks in group I, 1.8 weeks in group II, and 1 week in group III. Twelve infants required prolonged stay in the neonatal intensive care nursery because of prematurity. Six of 30 surviving infants had some abnormal physical findings at the time of initial discharge from the nursery. Premature rupture of membranes was the most common perioperative and postoperative complication. Chorioamnionitis developed in 17 of the 51 cases. Maternal complications included cervicovaginal fistula, deep vein thrombophlebitis, and pulmonary edema. PMID- 8240590 TI - A primary cytomegalovirus infection in pregnancy presenting with unusual hematologic manifestations: a case report. AB - A previously healthy multigravida woman presenting with pyrexia was found to be pancytopenic at 32 weeks' gestation. The leukopenia and the thrombocytopenia responded to supportive therapy. The anemia, which was Coombs' negative, continued and was associated with a drop in fibrinogen, but both eventually returned to normal with time. The isolation of the cytomegalovirus and the detection of an IgM immune response to cytomegalovirus implicated the virus as the agent responsible for the clinical manifestations. The fetus, who was well throughout the maternal illness, had no evidence of anemia or thrombocytopenia at birth; however, intrauterine infection was confirmed by isolation of the virus from the newborn's urine. PMID- 8240591 TI - Measurement of pulmonary capillary pressure during ritodrine tocolysis in twin pregnancies: a new noninvasive technique. AB - Five twin pregnant women were studied during treatment of preterm labor with ritodrine. Maternal pulmonary capillary pressure (PCP) and cardiac function were measured by noninvasive simultaneous recordings of the indirect carotid pulse, electrocardiography, phonocardiography, and M-mode echocardiography. Pulmonary capillary pressure significantly increased from the control value of 9 +/- 3.0 to 18 +/- 2.2 mmHg during the infusion period of therapy. Cardiac index and preejection period to left ventricular ejection time ratio were also increased during treatment with ritodrine. Our findings suggest that noninvasive measurement of PCP and cardiac function may be useful in monitoring twin pregnancies that develop cardiovascular side effects during tocolytic therapy. PMID- 8240592 TI - Pregnancy outcome following fetal exposure to tiaprofenic acid in the first trimester. AB - Fetal exposure to nonsteroidal inflammatory drugs beyond the 34th week of gestation is theoretically associated with premature closure of the fetal ductus arteriosus due to the inhibition of cyclooxygenase. Inadvertent exposure to tiaprofenic exposure in the first trimester is of concern due to the paucity of prospective human data. Experiential data are limited to retrospective reports submitted to the manufacturer and animal data, the extrapolation of which to humans is very inaccurate. We report pregnancy outcome in 12 women who voluntarily contacted the Motherisk Program in Toronto about first trimester exposure to tiaprofenic acid. These women were older than women who generally contact Motherisk (30.6 years +/- 3.9 years versus 29.9 years +/- 3 years). Three pregnancies ended in first trimester miscarriages and one woman elected to terminate her pregnancy. Of the eight live births, the mean gestational age at delivery was 39.2 weeks +/- 2 weeks, the mean birthweight was 3359 +/- 503 gm, no malformations were detected, and age of attainment of developmental milestones was normal. Although 25% of exposed cases ended in miscarriages, this may not be statistically meaningful. However, a similar trend is documented in animal studies and should be borne in mind when counseling the pregnant patient. PMID- 8240593 TI - Lactobacilli in Papanicolaou smears, genital infections, and pregnancy. AB - Papanicolaou (Pap) smears taken at the first antenatal visit of black African women were examined for lactobacillary expression and its correlation with the prevalence of genital infections and with the outcome of pregnancy. Lactobacillary morphotypes were absent (grade III) in 52% of 256 women. Trichomonas, Chlamydia, gonococci, or syphilis was identified in 54% of grade III women, and in 17% of grade I + II women (p < 0.0001). There were partially independent positive associations of Trichomonas, Chlamydia, and gonococci and a negative association of Candida morphotypes with grade III. Absence of lactobacilli, whether or not in the context of bacterial vaginosis, is an indicator for the presence of other genital infections. Birthweight was 2000 gm or less in 5% of 82 children of grade I + II mothers, and in 16% of 83 children of grade III mothers (p = 0.02). Thus, there are populations in whom the grading of lactobacilli in routine Pap smears helps to direct further investigations and to predict the outcome of pregnancy. PMID- 8240594 TI - Severe hemolytic disease of the newborn caused by anti-Gonzales antibody. AB - We report on a female infant with severe hemolytic disease of the newborn (HDN) secondary to anti-Gonzales antibody (anti-Go(a)) necessitating an exchange transfusion within the first day of life. The infant was born to a mother known to be Gonzales-antigen negative and a father who was Gonzales-antigen positive. The mother had an anti-Go(a) titer of 1:256 at 35 weeks' gestation. The infant was noted to have jaundice shortly after birth, with a bilirubin of 17.8 mg/dl (total) and .05 mg/dl (direct) at 11 hours of life. Coombs' test was positive and cord cells were Gonzales-antigen positive. Eluate on cord cells demonstrated anti Go(a). Despite aggressive phototherapy, the total bilirubin reached 23.3 mg/dl by 24 hours of age and a double-volume exchange transfusion was performed. Following the exchange transfusion, phototherapy was continued for several days. The hemoglobin, which was 19.8 gm/dl at 11 hours of age, remained stable through the hospitalization, and no further transfusions were required. History revealed that two prior pregnancies resulted in Gonzales-antigen positive infants. The first child experienced mild jaundice requiring no therapy, and the second child required phototherapy for prolonged hyperbilirubinemia. Previous reported cases of anti-Go(a) suggest that this is not a cause of severe HDN. However, in view of the current case, it may be prudent to follow women with anti-Go(a) prenatally with amniotic fluid bilirubin studies, serial antibody titers, and fetal hemoglobin levels. PMID- 8240595 TI - Outcome of term gestation neonates whose mothers received intrapartum antibiotics for suspected chorioamnionitis. AB - Intrapartum antibiotics are commonly given to women suspected of having chorioamnionitis. However, there is no consensus regarding management of their babies. To date, there are no data concerning the clinical courses and outcomes of term gestation, "pretreated" neonates. We performed a retrospective review to assess the clinical courses of such infants. From 1987 to 1989, chorioamnionitis was diagnosed in 123 women, 102 of whom received intrapartum antibiotics. Of the 86 term gestation infants born to women in the latter group, 63 were completely asymptomatic. When compared with these neonates, the 23 symptomatic infants were more likely to have lower median 1 and 5 minute Apgar scores (3 vs 8 and 7 vs 9, respectively; p < 0.0001), higher immature:mature neutrophil ratios on their initial complete blood counts (0.48 vs 0.24; p = 0.0003), and positive urine antigen studies for group B Streptococcus in the presence of negative blood cultures (4 vs 2; p = 0.042). The intrapartum courses of those infants destined to be symptomatic were more likely to be complicated by fetal tachycardia (p = 0.049) and malodorous amniotic fluid (p = 0.005). Since more than 95% of asymptomatic, pretreated, term gestation neonates have a benign clinical course, aggressive evaluation and management of these infants may not be necessary. PMID- 8240596 TI - Maternal and neonatal outcome associated with prolonged premature rupture of membranes below 26 weeks' gestation. AB - The outcomes of 41 patients with prolonged, premature rupture of membranes in the midtrimester (19.5 to 26 weeks) managed conservatively were retrospectively analyzed. The maternal obstetric history was notable for a high incidence of second trimester bleeding and a prior history of preterm delivery. The mean duration of ruptured membranes before delivery was 10.6 days and was unrelated to gestational age. Delivery occurred within 2 weeks in 75% of the cases. The major indication for delivery was chorioamnionitis in 71% of patients. The only major maternal morbidity was chorioamnionitis. The overall perinatal survival was 47%, but in infants exceeding 24 weeks' gestation or 500 gm weight, the survival increased to 75%. No significant limb abnormalities, facial anomalies, growth retardation, or pulmonary hypoplasia occurred in the study population. Long-term follow-up demonstrated that 28% of infants exhibited major neurologic or developmental deficits. PMID- 8240597 TI - Perinatal screening for drugs of abuse: reassessment of current practice in a high-risk area. AB - Anonymous urine toxicology screening among parturient women during 1 month in 1990 and selective newborn testing during this and the subsequent 4-month period was done to assess prevalence of drug use among parturients in a municipal hospital in the Bronx and to assess impact of infant urine toxicology screening on discharge placement. Infant testing was performed for maternal history of drug use, poor prenatal care (5 or fewer visits), or infant symptoms. Urine was screened for cocaine, opiates, methadone, barbiturates, amphetamines, and benzodiazepines. Of 204 women screened, 9.3% were positive. Of these, 74% were positive for cocaine and 21% revealed polysubstance use. Only 28.6% of cocaine positive mothers gave a history of use. Selective testing of 1196 newborns during this 5-month period revealed an apparent prevalence of cocaine exposure of 4.9%. Selective infant testing failed to identify 42.1% of newborns of cocaine-positive women. Social work evaluation was performed on all families and was the basis for reporting to state agencies for protective services. Only 6 of 83 drug-positive infants entered foster care, none because of positive toxicology per se. Selective infant toxicology studies miss many cocaine-exposed infants and has little impact on placement. Universal social work evaluation of families may be as effective and freer of bias than selective urine screening. PMID- 8240598 TI - Three cases of Haemophilus influenzae amnionitis. AB - Haemophilus influenzae is an uncommon cause of obstetric and neonatal infections. Three cases of amnionitis caused by H. influenzae are presented. These cases occurred in three maternal transports to our tertiary level hospital within an 11 day period. The organisms were analyzed for biotypes, penicillinase status, and composition of membrane fatty acids. PMID- 8240599 TI - Acute maternal renal insufficiency in premature labor treated with indomethacin. AB - Three cases of acute renal insufficiency in pregnant women who were treated with indomethacin for premature labor are reported. At the time of presentation, all three women had normal renal function but within 30 hours of indomethacin therapy they were noted to have significant decreases in urine output and rising serum creatinines. The average time to recovery of renal function was 5 days. A consistent feature in all three women was the development of dyspnea associated with hypoxemia. PMID- 8240600 TI - Feasibility of postpartum rapid hospital discharge: a study from a community hospital population. AB - We have examined the safety and efficacy of an early postpartum discharge program in 289 patients who were identified prior to delivery and counseled and educated for discharge 12 to 36 hours after delivery. A total of 55% of the study patients were able to be discharged early. There were 4.3% significant maternal problems and 3% significant neonatal problems identified in the first 72 hours after delivery by a nurse practitioner home visit. Significant maternal problems after 72 hours (10%) and neonatal problems (5%) were also found by follow-up visits. The hospital readmission rate was 1.8%. These incidences are consistent with other studies in the literature and suggest that some type of early follow-up of both the mother and infant should routinely accompany early postpartum discharge programs. PMID- 8240601 TI - A comparison of sonographic growth curves of triplet and twin fetuses. AB - Sonographic growth curves of 47 normal triplet pregnancies were compared with those of 71 uncomplicated twin gestations. Starting from the 25th week, the biparietal diameter in triplets was found to lag progressively compared with that of twins, reaching a mean difference of 2 weeks at 36 weeks gestation. Similarly, a significant difference was found between the femur length and head to abdomen circumference ratio growth curves of triplets and twins. It is concluded that as pregnancy continues from the 25th to the 36th gestational week, there is a significant delay in the growth patterns of normal triplet pregnancies compared with twin gestations. PMID- 8240602 TI - Ionized serum magnesium levels in umbilical cord blood of normal pregnant women at delivery: relationship to calcium, demographics, and birthweight. AB - Using a novel ion-selective electrode for ionized Mg (IMg2+), we sought to: (1) determine levels and fractions of IMg2+ in umbilical vessels of normal pregnant women: (2) determine their relationships with ionized calcium (ICa2+) levels: (3) determine whether any demographic or clinical parameters affect these levels of divalent cations: and (4) compare levels of IMg2+, total Mg, percent IMg2+, and ICa2+ with sera of normal pregnant and nonpregnant women. We obtained umbilical arterial and venous serum from 38 normal subjects and venous samples from 26 normal pregnant women and 42 nonpregnant women. The mean umbilical venous IMg2+ level (0.51 +/- 0.01 mmol/liter) was significantly greater than the umbilical arterial level (0.48 +/- 0.01 mmol/liter, p < 0.05), and the latter was similar to maternal venous levels (0.48 +/- 0.01 mmol/liter). The maternal venous mean value is 20% less than that found (0.60 +/- 0.005 mmol/liter) in peripheral serum of nonpregnant women. The ionized fractions were significantly (p < 0.01) higher in the umbilical veins than umbilical arteries, and the percent IMg2+ fractions were similar in umbilical arteries and maternal venous blood. IMg2+ levels were lower in multiparous patients and those who received epidural analgesia and higher in patients who had operative vaginal delivery and in Asians. Ionized fractions of Mg in umbilical veins were higher in large for gestational age infants. Our findings provide evidence of a Mg deficiency in pregnancy and suggest that measurement of IMg2+ may have clinical value for diagnosis and therapy of such deficiency.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240603 TI - Sonographic documentation of uterine retroversion mimicking uterine sacculation. AB - Uterine retroversion during the first trimester is quite common. However, as the uterus increases in size, self-correction usually occurs and the second trimester pregnant uterus becomes an abdominal organ. If anteversion does not occur, pelvic impaction will result. The prevalence of late first and early second trimester uterine impaction is approximately 1 in 3000 pregnancies. Our case documents sonographically the spontaneous resolution of a markedly retroverted uterus at 26 weeks' gestation. PMID- 8240604 TI - Pregnancy-induced hypertension and early neonatal death: a case-control study. AB - Literature on the association between pregnancy-induced hypertension and early neonatal death is limited. The present case-control study consisted of 342 early neonatal deaths and 523 frequency-matched controls. After controlling for other factors such as infant sex, gravidity, maternal age, threatened abortion, and maternal anemia, moderate to severe pregnancy-induced hypertension was associated with an increased risk of early neonatal death with apparent dose-effect gradients (odds ratio = 1.8 for moderate and 2.2 for severe). The biologic plausibility of pregnancy-induced hypertension as a cause of neonatal mortality is discussed. PMID- 8240605 TI - Congenital cytomegalovirus infection presenting as cerebral ventriculomegaly on antenatal sonography. AB - Antenatal sonography in two cases demonstrated cerebral ventriculomegaly and decreased head circumference, subsequently found to be secondary to cytomegalovirus (CMV) infection. In both cases, the lateral ventricles were enlarged and lissencephaly was diagnosed in the neonatal period. The findings of ventriculomegaly and decreased head circumference on antenatal sonography warrant further investigation for CMV via amniotic fluid cultures or fetal blood, given the poor prognosis in infants with symptomatic infection. PMID- 8240606 TI - Out-of-hospital health care: innovation and excellence for the new century. PMID- 8240607 TI - Crisis in primary care: will midwives meet the challenge? PMID- 8240608 TI - Do cabbage leaves prevent breast engorgement? A randomized, controlled study. AB - A randomized, controlled trial was conducted to evaluate the effect of cabbage leaves on mothers' perceptions of breast engorgement and the influence of this treatment on breastfeeding practices. The subjects, 120 breastfeeding women 72 hours postpartum, were randomly allocated to an experimental group who received application of cabbage leaves to their breasts, or to a control group who received routine care. The experimental group tended to report less breast engorgement, but this trend was not statistically significant. At six weeks, women who received the cabbage leaf application were more likely to be breastfeeding exclusively, 76 and 58 percent (35/46 vs 29/50; P = 0.09), and their mean duration of exclusive breastfeeding was longer (36 vs 30 days; P = 0.04). The greater breastfeeding success in the experimental group may have been due to some beneficial effect of cabbage leaf application, or may have been secondary to reassurance and improved confidence and self-esteem in these mothers. PMID- 8240609 TI - Expectations and experiences of pain in labor: findings from a large prospective study. AB - Data on expectations and experiences of pain in labor are presented from a prospective study of over 700 women who gave birth in six maternity units in southeast England. Most women preferred to keep drug use to a minimum, even though they expected labor to be "quite" or "very" painful. The ideal of avoiding drugs was unrelated to education or social class. Women who preferred to avoid drugs were more likely to do so, and were more satisfied with the birth overall than women who used drugs. In general, women tended to get what they expected. Breathing and relaxation exercises were widely used, and were most successful for those who had expected them to be so. Anxiety about the pain of labor was a strong predictor of negative experiences during labor, lack of satisfaction with the birth, and poor emotional well-being postnatally. PMID- 8240610 TI - The squatting position for the second stage of labor: effects on labor and on maternal and fetal well-being. AB - A cohort study was designed to assess the effects of maternal squatting position for the second stage of labor on the evolution and progress of labor, and on maternal and fetal well-being. Outcomes from 200 squatting births, randomly selected from a sample of 1000, were compared with 100 semirecumbent births, randomly selected from a sample of 300. Data collection was by chart review. The two groups were similar with respect to most antepartal, intrapartal, and socioeconomic variables likely to affect labor outcomes. The mean length of the second stage of labor was 23 minutes shorter in squatting primiparas and 13 minutes shorter in squatting multiparas than in semirecumbent women. Squatting women required significantly less labor stimulation by oxytocin during second stage (P = 0.0016), and they showed a trend toward fewer mechanically assisted deliveries. Significantly fewer and less severe perineal lacerations occurred, and fewer episiotomies were performed in the squatting group (P = 0.0001). No statistically significant differences were found between groups for third-stage complications and infant complications. PMID- 8240611 TI - Immersion in water during labor and birth: the need for evaluation. AB - Despite widespread and increasing use of immersion in water during labor, birth, or both in many different countries, no reliable information is available about its advantages, hazards, and resource implications. We describe the development of immersion in water during labor and birth, the current use of this form of care in the United Kingdom, the research that has been conducted to date, and possible strategies for future research. Three approaches to research are suggested: a survey of existing practice, a randomized controlled trial, and a confidential register of serious adverse events. PMID- 8240612 TI - Redefining women's work during pregnancy: toward a more comprehensive approach. AB - The potential effects of work and activity on the processes and outcomes of pregnancy should be assessed when a woman's care plan is developed. Perinatal caregivers are limited by a narrow definition of work that encompasses only employment. This commentary expands the description of women's work during pregnancy, and proposes the addition of some activities not currently considered in most prenatal assessments, including home care responsibilities, the maintenance and development of intimate relationships, and the tasks identified with becoming a mother. All this has implications for research, clinical practice, and public and health policy. PMID- 8240613 TI - Commentary: whose work is it, then? PMID- 8240614 TI - Active management: a continuing benefit in nulliparous labor. PMID- 8240615 TI - The active management of labor: whose agenda? PMID- 8240616 TI - Active management of labor: not the answer to dystocia. PMID- 8240617 TI - Reanalysis of a multireported trial on home uterine activity monitoring. AB - A randomized multicenter study of home uterine activity monitoring using the "Term Guard," is widely cited in support of the Term Guard monitoring system. Critical appraisal of the data in the six reports of the study showed that all reports scored poorly on avoidance of allocation, exclusion, and assessment bias. In all of them the numbers of women for whom outcomes were reported were different, and none provided information on infant outcomes. Comparison of analyses and subanalyses showed inconsistencies that defied the laws of arithmetic. Reanalysis of the data, as well as could be done from the available information, showed that use of the Term Guard system was compatible with both an increase and a decrease in the frequency of preterm delivery (95% CI +2.2% to 16.5%). It is concluded that the reports of the multicenter study contain such high potential for bias and so many internal inconsistencies that they should be dismissed entirely. PMID- 8240618 TI - Attenders and nonattenders at childbirth education classes in Australia: how do they and their births differ? AB - This study assessed the associations between attendance at childbirth preparation classes and the health behaviors, birth events, satisfaction with care, and later emotional well-being of women having their first child. A postal survey was conducted of a population-based cohort of 1193 women who gave birth in two weeks in 1989 in Victoria, Australia. The response was 71.4 percent (790/1107). Classes were attended by 245 (83.9%) of 292 primiparous women. Those who did not attend were significantly more likely to be under age 25 years, not to have completed secondary education, to be single, to have a low family income and no health insurance, and to be public hospital clinic patients. Differences between women who attended classes and those who did not, with respect to measures of pain and to the use of procedures, interventions, and pain relief, were rare and minor. No differences occurred between the groups in their satisfaction with the provision of information through pregnancy, birth, and the postnatal period. Only one of five measures of satisfaction with care was less favorable in nonattenders. Attenders were not more confident about looking after their infants at home or less likely to be depressed eight months after birth. Significant differences occurred between the groups on four health behaviors: cigarette smoking, missed antenatal appointments, breastfeeding, and alcohol consumption during pregnancy. Attendance at childbirth preparation classes in Victoria is not associated with differences in birth events, satisfaction with care, or emotional well-being among women having their first child. PMID- 8240619 TI - Effects of prenatal classes cannot be measured by obstetric management. PMID- 8240620 TI - Accuracy of Leopold maneuvers in screening for malpresentation: a prospective study. AB - This study describes the accuracy of Leopold maneuvers as a screening procedure for fetal malpresentation. The frequency of fetal malpresentation ranges from 15 percent at 32 weeks to 4 percent at term. We prospectively determined fetal presentation by performing Leopold maneuvers on 150 women, followed by a fetal ultrasound examination for comparison. Experienced certified nurse-midwives performed the maneuvers with high sensitivity (88%), specificity (94%), positive predictive value (74%), and negative predictive value (97%) in a population with a 17 percent frequency of fetal malpresentation. We conclude that the maneuvers used by experienced clinicians can be effective as a screening tool for fetal malpresentation, particularly in settings where ultrasound may not be readily available. PMID- 8240621 TI - Maternal recall and medical records: an examination of events during pregnancy, childbirth, and early infancy. AB - It is not currently known whether sufficiently precise data on a previous pregnancy, labor and delivery, and early infancy can be obtained retrospectively. We conducted a telephone survey in 1991 of women who delivered babies between 1984 and 1986 at two teaching hospitals in Nashville, Tennessee, to assess how well mothers recall information on factors predictive of an adverse birth outcome. The survey yielded 102 usable responses that were compared with hospital records for mothers and infants. Overall, 89 percent agreement was found between women's responses and their charts. Respondents were not reluctant to answer potentially sensitive questions, and their technical knowledge was typically better for their own health than about some prenatal diagnostic procedures and their infants' health. We found no difference in recall accuracy according to whether mothers experienced some adversity with the index pregnancy. Accurate perinatal information can generally be obtained with a recall period as high as four to six years. PMID- 8240622 TI - "What are they talking about? Is something wrong?" Information sharing during the second stage of labor. AB - Twenty postpartal women were shown videotapes of their second stage labors and simultaneously interviewed. During separate interviews, their 25 caregivers were also shown the videotapes and interviewed. The interviews were analyzed for major themes, one of which was sharing information during labor. Although women and caregivers appeared to agree about what information laboring women require and how it should be given, caregiver perceptions of the quality of their information giving were more positive than mothers' perceptions. Many women wanted more informational support, especially in alleviating unvoiced fears about their baby's health. PMID- 8240623 TI - A plan for implementing mother-baby nursing. AB - This article describes the plan and implementation for converting from traditional obstetric nursing to a mother-baby care delivery system in a level 2 hospital in upstate New York. It identifies the pitfalls, problems, and process to follow for the successful implementation of mother-baby nursing. PMID- 8240624 TI - Methodologic issues in assessing the active management of labor. PMID- 8240625 TI - Effective control or effective care? PMID- 8240626 TI - The active management of physicians. PMID- 8240627 TI - A final comment ... managing the uterus, the woman, or whom? PMID- 8240628 TI - The final fatal blow to routine episiotomy. PMID- 8240629 TI - The lesbian childbearing couple: a case report. PMID- 8240630 TI - [Function preserving laryngeal cancer surgery in Hungary-- a historical retrospect and 25 years personal experiences]. AB - The continuity in the trends in conservation and surgery in the Hungarian laryngology are emphasized. The first Hungarian book on this topic, "Diseases of the Larynx" by Navratil, was published 127 years ago ("A gegebajok", Pest, 1866). Onodi popularized the laryngectomy and the results of the surgery of laryngeal carcinoma in the first two decades. In 1900, Herczel described on a vertical partial laryngectomy. Lenart was an internationally accepted Hungarian surgeon in laryngology whose results (cordectomy 61%, partial laryngectomy 48%, hemilaryngectomy 42%, total laryngectomy 35%) were consistently successful. Rethi was a famous name in the recent period of Hungarian laryngeal surgery for laryngotracheal stenosis and laryngeal carcinoma. In the past 25 years, the occurrence of supraglottic cancer was the highest. The author reports his personal results and stresses the importance of conservation surgery. Of a total of 643 patients with supraglottic carcinoma, 126 (20%) were treated by primary radiotherapy, 169 (26%) were operated on by total laryngectomy, 245 (38%) by supraglottic laryngectomy, 61 (9.5%) by supraglottic subtotal laryngectomy (personal method) and 42 (6.5%) by resection of the supraglottic larynx and the base of the tongue. Local tumour control was constantly good with a local recurrence rate of 6.6%. The 5-year survival rate was 74.3% for supraglottic laryngectomy, 68.9% for subtotal laryngectomy and 62% for the resection of the supraglottic larynx and the base of the tongue group. PMID- 8240631 TI - [Functional and morphologic results of pedicled and microvascular anastomosis in tissue transfer]. AB - From 1982 to 1992, large intraoral and pharyngeal soft-tissue defects were reconstructed using myocutaneous island flaps in 95 patients, free jejunal interpositions in 10 patients and forearm flaps in 18 patients at the Magdeburg School of Medicine, Department and Clinic of Otorhinolaryngology. Sixty-five patients were followed up after various time periods in order to assess the functional results. Investigation and evaluation of the individual patients' resultant quality of life were based on a specifically prepared examination questionnaire. In particular, the evaluation included: the mouth opening gap, motility of the tongue, sufficient masticatory area, adequate provision to fit artificial teeth, malocclusion, the extent of mandibular deviation, assessment of the swallowing act, articulation disturbances, paraesthesia, function accessory nerve, atrophy of the trapezius muscle, motility of the cervical spine, flexibility in the shoulder girdle, as well as subjective information obtained from the patients (pain, feeling of dryness, increased salivation, occupational rehabilitation, organisation of leisure time). In 45 patients, biopsy material from the transferred tissue was examined at varied time intervals after primary therapy. IN CONCLUSION: The functional results obtained for the myocutaneous island flaps, the forearm flap and the free jejunal interposition were satisfactory, and only minor differences were noted. From the morphological point of view, all the three types of tissue transfer were found to have adapted adequately to the recipient site. PMID- 8240632 TI - [Functional and cancerologic aspects in the use of jejunum in the ENT area]. AB - In this article, we report about the experiences at our ENT-University Clinic in Vienna in 76 patients with free transplanted microvascularly anastomosed autologous jejunum used for reconstruction following extensive resection of mainly stage IV carcinomas in the head and neck. Jejunum was inserted as a siphon like tube between the trachea and the hypopharynx in 43 patients primarily for restoration of speech (in 10 cases the pharynx was also reconstructed). In 33 patients, the upper digestive tract was reconstructed, as either patch or tube. They were then evaluated according to functional results of speech and swallowing, aesthetics as well as 5-year survival rates. We achieved good results in 35 speech patients (81.4%) and in 22 swallowing patients (66.6%). The aesthetic function was influenced favourably because the volume of the jejunal transplant fills up the defect in the neck after resection and creates a tolerable neck-shape. The 5-year survival of all available 54 patients was 26%, patients for speech restoration 31.7% and patients for swallowing restoration 17.5%. These results are to be considered under the aspect that patients with swallowing restoration primarily had more severe disease. Complications such as like perioperative mortality and total necrosis can be avoided by careful evaluation of the patients to be operated on. With the experiences gained from this study, we now have exact indications for the use of jejunum grafts and thus can expect very high rates of success. PMID- 8240633 TI - [Palpation and sonography in after-care of head-neck tumor patients: comparison of ultrasound tumor entity parameters]. AB - The ranking of postoperative palpation and sonography is examined in respect of imaging of neck lymph node enlargement and neck lymph node metastases or lymphomas, using a prospective study involving 127 patients whose head and neck tumours had already undergone treatment. These patients had been subjected to tumour aftercare for an average period of 26 months. Sonography is well suited for detecting the presence of enlarged neck lymph nodes, the accuracy being 97.5%. In this study palpation was clearly less safely defined, the accuracy being only 85%. The following results were obtained in respect of the specific imaging of metastases: sensitivity of palpation (75%) is below that of sonography (M/Q ratio) (93%). In respect of specificity palpation also yielded poorer results than sonography (M/Q ratio 94%) because of the relatively high proportion of false positive palpations (17%). The reason for this is non-differentiation between reactively enlarged lymph nodes and lymph nodes metastases. The advantage of sonography vs palpation by using the M/Q ratio is therefore the accuracy of the findings (94% sonography, 81% palpation) and hence the clear reduction in false positive and false negative findings. Furthermore, sonography can image the exact relationship to surrounding tissue and especially the vessels. The drawback of sonography is that lymph nodes < 9 mm maximal diameter cannot be assessed with the safety and accuracy mentioned above. Sonomorphological criteria such as the central marrow reflex and a widened lymphatic node cortex enhance diagnostic safety of the sonographic expansion parameters to yield an assessment of tumour status. PMID- 8240634 TI - [Indications and value of various techniques of nasal alar reconstruction]. AB - Reconstruction of the ala of the nose is a very common task for the plastic surgeon. Many different techniques are available, but as in all reconstructive procedures in the face it is important to consider the combination of function and aesthetics. It is not only important to close the defect, but how it looks afterwards. According to these criteria the various methods were analysed and local flaps were found to be the most favorable technique due to their identical texture and colour match to the surrounding tissue. On the nose, the skin of the tip and the ala is very tight which makes it difficult to use local flaps there. However, there is a surplus and looseness of skin in the glabella region. This skin can be transferred by an axial fronto-nasal flap, pedicled on a branch of the angular artery as described by Marchac. With this flap any point of the nose can be reached. In our opinion, this flap appears preferable to all others. The new technique of ala reconstruction based on this axial fronto-nasal flap is demonstrated. The skin defect is covered by this flap and for the inner lining a mucosal flap from the septum can be used which includes a strip of cartilage for stabilisation. In addition the surplus of tissue at the point of rotation, which normally has to be resected as a Burow's triangle, can also be used for the inner lining. PMID- 8240635 TI - [Reconstruction of lacrimal ducts]. AB - For reconstruction of lacrimal drainage after injury of tumour resection indications, surgical technique and results of five surgical procedures are presented: primary suture with splinting, conjunctivorhinostomy, endonasal dacryocystorhinostomy, Heermann tube and mobilisation of nasolacrimal duct. Performing these techniques, solitary or in combination, one achieves almost a good lacrimal drainage. The results of our retrospective study are based upon a combination of subjective and objective findings. Success rate was 96% for direct suturing of the tear ducts with intubation, 100% for conjunctivorhinostomy, 96% for endonasal dacryocystorhinostomy in case of postsaccal stenosis, 83% for Heermann/Jones tube and 100% for relocation of the nasolacrimal duct. PMID- 8240636 TI - [Isolated medial orbital wall fractures: results of minimally invasive endoscopically controlled endonasal surgical technique]. AB - Between 1991 and 1993, 9 cases of isolated medial orbital wall fractures were diagnosed with 8 patients, 8 of them complaining of horizontal diplopia due to medial rectus muscle entrapment. An endoscopic-controlled endonasal surgical procedure was performed in 7 patients. The endonasal procedure provided adequate exposure for removal of the incarcerated tissue as well as covering the defect. All 7 patients had good results with no restriction of lateral gaze and There were no surgical complications observed. The first step of the endonasal technique is a partial resection of the middle turbinate. During the following partial ethmoidectomy, the fracture of the ethmoid and medial orbital wall are exposed. This must be done carefully while avoiding any trauma to the herniated orbital contents. After repositioning the orbital fatty tissue, lyodura is introduced and fixed with fibrin glue. For larger defects, free graft of mucosa is recommended. The nasal packing is done between two silicon sheets in order to simplify the removal after 8 days. All 7 patients proved to be free of motility disturbances after the intervention. The endonasal method is limited to isolated medial wall fractures; in all other cases of midfacial fractures combined techniques have to be used. PMID- 8240637 TI - [Foreign bodies of the esophagus and upper gastrointestinal tract in childhood]. AB - In a retrospective study we evaluated 49 endoscopies performed under suspicion of a foreign body in the oesophagus or upper gastrointestinal tract. In 27 cases a foreign body was easily removed. One child had to undergo a cervical oesophagotomy as a result of a chronically incarcerated foreign body. Because of the very low complication rate of endoscopy and a much higher risk of complications due to unknown foreign bodies, early endoscopic diagnosis is recommended even if the foreign body is only suspected. PMID- 8240638 TI - [Rational diagnosis of dysphagia]. PMID- 8240639 TI - [Current status of sialography in salivary gland diseases]. PMID- 8240640 TI - [The effect of flumazenil on the endocrine stress reaction following modified neuroleptanesthesia]. AB - This study was undertaken to investigate in a prospective randomized way the influence of the benzodiazepine antagonist flumazenil on endocrine stress response and haemodynamic parameters after modified neuroleptanaesthesia. A total of 24 patients (ASA scores I or II) aged between 18 and 60 who were scheduled for major gynaecological surgery, were investigated. For modified neuroleptanaesthesia, midazolam, fentanyl and vecuronium were administered in standardized doses. After extubation, patients of the flumazenil group received initial injections of 0.2 mg flumazenil to antagonize the residual effect of midazolam and additional doses of 0.1 mg per minute until the desired level of vigilance was reached (awareness of person, time and place). In the control group no flumazenil was used. Endocrine stress parameters and haemodynamic parameters were measured at 7 different times, from before induction of anaesthesia up to 60 minutes after the operation. In both groups, a marked increase in endocrine stress response was observed. Adrenaline, noradrenaline, antidiuretic hormone, adrenocorticotropic hormone, cortisol, glucose and lactate, however, were not additionally influenced by the antagonism. No influence of flumazenil on mean arterial pressure, heart rate and arterial oxygen saturation was observed. After modified neuroleptanaesthesia, a careful antagonism of midazolam with small doses of flumazenil is not disadvantageous with respect of endocrine stress response and haemodynamic reactions. PMID- 8240641 TI - [Soda lime--service life, consumption and costs in relation to fresh gas flow]. AB - The utilisation time of carbon dioxide absorbers in anaesthesia textbooks is nearly identically specified to last about 5 hours. Therefore in most departments the soda lime of the absorbers is changed routinely on a daily schedule. As rebreathing volume increases considerably with fresh gas flow reduction, the question arises as to whether the soda lime should be changed at even shorter intervals--if low-flow anaesthesia is performed routinely--to to guarantee carbon dioxide absorption safely. In three anaesthesia machines with different technical properties the carbon dioxide absorbers were only changed when the inspiratory carbon dioxide concentration increased to 1 vol. %, indicating definite exhaustion of the soda lime. If a fresh gas flow of 4.4 l/min was used exclusively, utilisation times of 42.7 and 62.3 hours were measured using absorbers filled with 1 l of soda lime pellets, whereas a utilisation time of 98.7 hours was noted with a 1.5 l absorber, all values being surprisingly higher than the above-mentioned figure. If, however, minimal flow anaesthesia is performed routinely in clinical practice, the percentage of time in which the fresh gas flow can really be reduced to 0.5 l/min does not exceed 50 to 80%. Under these conditions the utilisation time of the absorbers decreases to between one half (1/2) and one quarter (1/4) of the utilisation time which can be gained if a flow of 4.4 l/min is used. Thus, the performance of minimal-flow anaesthesia increases the consumption of soda lime two- to fourfold.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240643 TI - The measure of change. PMID- 8240642 TI - [Anesthesia for renal transplantation: continuous neuromuscular blockade with atracurium and the management of intraoperative hypertension with nifedipine]. AB - Patients undergoing kidney transplantation often suffer from essential hypertension and coronary artery disease, for which perioperative treatment with nifedipine proved to be effective. If calcium-channel blockers and nondepolarizing muscle relaxants are used simultaneously, their synergistic effect at the neuromuscular cleft must be considered. On the other hand because of its extrarenal elimination no significantly altered effects are expected for patients with terminal renal failure. This prospective study comprised 30 patients undergoing kidney transplantation who were 2 kg over normal weight after a preoperative dialysis and infusion of lactated Ringer solution. Fifteen minutes after introduction of balanced anaesthesia with isoflurane, nitrous oxide and fentanyl, patients were assigned to the treatment group (N) with hypertension (mean arterial pressure (MAP > 110 mmHg) and subsequent management with nifedipine or to the control group (0). Treatment was aimed at keeping MAP between 90 and 110 mmHg. The neuromuscular status was electromyographically assessed by the train-of-four-principle. There were evaluated the duration of action of the initial dose of atracurium (0.5 mg/kg) from injection time to T1 = 2% (WZ 2), the dose of atracurium for a continuous neuromuscular blockade (DD 98 in mg/kg/h), the recovery time and the recovery index. Fourteen patients with hypertension received a bolus of 10 micrograms/kg nifedipine. A significant reduction in blood pressure (p < 0.05) to the desired range was achieved by a subsequent infusion of nifedipine of 5 to 40 micrograms/kg/h. In 16 patients MAP was kept between 90 and 100 mmHg without any additional therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240644 TI - The influence of teeth on denture base processing accuracy. AB - The Michigan Computer Graphics Coordinate Measuring System was used to determine the influence of denture teeth on the accuracy of processed denture bases. Three different processing methods--compression molding, injection pressing, and vacuum assisted resin pouring--were used. The presence of denture teeth was determined to affect the accuracy of the processed denture bases of all three techniques. For denture bases without teeth, the compression-molding technique demonstrated significant differences in accuracy; however, no significant differences were found in the injection-pressed or resin-poured denture bases. PMID- 8240645 TI - The prognostic value of denture complaints. AB - This study tested whether information obtained from an inventory of denture related variables can help a dentist recognise the potentially slowly adapting denture patient. Sixty-one patients completed an inventory of denture complaints prior to the fabrication of complete maxillary and mandibular dentures. Subjects were subsequently divided into groups of slowly adapting and rapidly adapting patients. A clinically derived classification was compared to a theoretic model. Seventy-four percent of the theoretically slowly adapting patients and 89% of the theoretically rapidly adapting patients corresponded with their clinical classifications. PMID- 8240646 TI - Three-dimensional analysis of the casting accuracy of one-piece, osseointegrated implant-retained prostheses. AB - The three-dimensional distortions of implant frameworks of two designs, reflecting the early "Toronto" U-shaped design and a current L-shaped design, were measured. A conventional commercial laboratory one-piece casting method, using a silver-palladium alloy, was used. Distortion was defined by five displacement variables for the centroids of each of five gold cylinders incorporated in each casting: three orthogonal translational displacements (dx, dy, and dz) and two rotational displacements (d theta x and d theta y). A computerized coordinate measuring machine was used. Overall translational displacements measured were -17.4 +/- 15 microns for dx, 6.1 +/- 7.1 microns for dy, and -0.1 +/- 1.7 microns for dz. The overall global translational displacement, dR, was 20.2 +/- 14.5 microns. Overall rotational displacements were -0.098 +/- 0.109 degrees for d theta x and -0.020 +/- 0.097 degrees for d theta y, giving an average absolute angular change, d theta R, of 0.158 +/- 0.077 degrees. No significant differences were found between U- and L-shaped castings, except for dy values for cylinders 2, 3, and 4, and d theta x values for cylinder 1. Cylinder location within the arch was associated with a specific direction and magnitude of translational and rotational displacement. Even small rotational displacements may manifest large gap appearances with the one-screw test because of the "moment arm" effect. However, other rotational displacements may be hidden, depending on the direction of tilt and the moment arm length. PMID- 8240647 TI - Effect of light-tip distance on polymerization of resin composite. AB - The inability to place a light tip in close approximation to a resin restoration may affect the resultant polymerization and clinical durability of the restoration. This research measured light intensity at the surface of a resin composite, as well as 2 mm within its bulk, as the tip-to-resin distance is moved from 0 to 10 mm. The polymerization of the resin composite at both locations was measured for the various tip distances using exposure durations of 10, 20, 40, and 60 seconds. Light intensity did not decrease with the inverse square of the tip distance. The polymerization on the surface was greatly dependent upon the duration of exposure. The extent of polymerization 2 mm below the surface was still dependent primarily upon exposure duration, but intensity had a significant affect. For exposure durations of 10, 20, and 40 seconds, a tip distance greater than 4 mm demonstrated a significant decrease in resin polymerization 2 mm below the resin composite surface. PMID- 8240648 TI - Using computerized cephalometrics to analyze the vertical dimension of occlusion. AB - A computer program that uses cephalometric analyses for determining the patient's occlusal vertical dimension has recently been introduced. Data generated from this computer implies changes in incisal-pin position for articulated casts. This study evaluated the accuracy of this vertical-dimension program using 24 completely dentate, white male subjects with clinically acceptable occlusal vertical dimensions. A cephalometric radiograph was made, and measurements from the tracing were entered into the computer for analysis. Recommended incisal-pin changes ranged from -11 to +25.3 mm, with a mean change of 8.4 mm for all methods tested. These results showed a low correlation with each subject's clinically determined occlusal vertical dimension. PMID- 8240649 TI - Accuracy of replacing three tapered transfer impression copings in two elastomeric impression materials. AB - The accuracy of replacing three manufacturers' (Dental Imaging Associates, Implant Innovations Inc, and Steri-Oss) tapered transfer impression copings into impressions made with two different materials (Impregum F and Extrude) was evaluated. Five operators replaced these copings into the elastomeric impression materials. The angular deviations (replacement error) of the copings between the control (original) position and the replaced position in the impressions were evaluated. It was found that no one coping could be replaced into the impressions accurately and consistently by all five operators. However, there were significant differences in replacement accuracy between the three copings. Lower angular deviations were found for the Steri-Oss and Dental Imaging Associates copings when compared to the Implant Innovations coping. There was no significant difference in replacement accuracy between the two impression materials. PMID- 8240650 TI - Physical properties of fixed prosthodontic, resin composite luting agents. AB - This study determined and compared physical properties of six fixed prosthodontic resin composite luting agents and one control. Inorganic filler content, compressive strength, diametral tensile strength, film thickness, and Knoop hardness were determined. The amounts of filler and the physical properties varied widely among materials. One material, which contained a minimal amount of filler, underwent so much plastic deformation that its strengths could not be measured. All other materials demonstrated high strengths. The materials with the least filler demonstrated the least resistance to indentation. The material with the lowest film thickness had not set at the time of measurement, 10 minutes after mixing. Most materials had unacceptable film thicknesses. Only one material demonstrated acceptable physical properties throughout the study. PMID- 8240651 TI - The effect of finishing and polishing on surface roughness of a processed resilient denture liner. AB - A raised ridge on a Molloplast-B sample was removed using one of three burs, four stones, or a coarse sandpaper disc. Samples were polished using pumice alone, tin oxide alone, or both agents. A flat, unfinished sample served as a control. The average surface roughness was measured after reductions and subsequent polishings. The length of time required for initial reduction was recorded. Average surface roughness measurements ranged from 16.0 to 1.4 microns, with the control measuring 0.66 micron. Without polishing, burs produced rougher surfaces than stones and required longer times for reduction. Bur samples also remained rougher than stone samples after pumicing. No significant differences were found between treatments or controls after the use of either a combination of pumice and tin oxide or tin oxide alone. PMID- 8240652 TI - The effect of die spacing on crown deformation and seating time. AB - A machined brass crown, onto which a strain gauge had been attached, was filled with a silicone fluid and placed on various dies to simulate luting using zinc phosphate cement. Space between the crown and the die was varied, as was the seating force. As the load increased, crown deformation increased and time to seat decreased. Increasing the axial space decreased the time to seat but did not affect the strain recorded for a given load. PMID- 8240653 TI - Condylar position recorded using leaf gauges and specific closure forces. AB - Retruded interocclusal records were made for 40 subjects after deprogramming using leaf gauges and controlled incisal forces, which were exerted on a specially constructed occlusal force sensor. These records were used to assess the resulting displacements of the mandibular condyles from their positions in centric occlusion. The leaf gauges were found not to position the condyles inferiorly and posteriorly as has been previously reported. PMID- 8240654 TI - Implant information and insights. PMID- 8240655 TI - Bis-naphthalimides: a new class of antitumor agents. AB - A series of bis-intercalators bearing the 1,8-naphthalimide chromophore has been synthesized and in vitro activities determined. Most compounds have higher activities in HT-29 than the leader compounds Mitonafide and Amonafide. One of them (22) was selected for in vivo studies and presents an interesting activity in MX-1 and OVCAR models. (22) also acts as antitopoisomerase II. PMID- 8240656 TI - The DNA complexes of 2,5-diaziridinylbenzoquinone and 3,6-dimethyl-2,5 diaziridinylbenzoquinone. A molecular mechanics study. AB - Computer modeling has been used to study the non-covalent interaction of the quinone and hydroquinone forms of 2,5-diaziridinyl-1,4-benzoquinone (DZQ and DZHQ, respectively) and its 3,6-dimethyl derivative (MeDZQ, MeDZHQ) with DNA. Calculations were performed using DNA duplexes containing 5'-TGCA and 5'-TGTC binding sites. The results allow a rationalization of the sequence selectivity of DNA alkylation exhibited by these ligands. For DZHQ a preference for binding to the TGCA site, over the TGTC site, was predicted for a range of diverse ligand/DNA orientations. The ligand preferentially adopts a position orthogonal to the base pair planes. The complex is stabilized by guanine N7 to ligand OH hydrogen bonds at the GC step. The stability of the TGTC complexes of DZHQ is reduced by a clash between the thymine methyl of the GTC sequence and the ligand. In MeDZHQ/TGCA complexes, a similar clash causes the aromatic ring plane to rotate about 45 degrees from the base pair planes. The most stable orientation of the MeDZHQ/TGTC complex has the ligand symmetrically disposed about the T-A base pair in the center of the GTC site, with hydrogen bonds formed between both ligand hydroxyl groups and guanine N7 atoms on opposite strands. For all four complexes, a secondary energy minimum was located for an orientation in which the face of the aromatic ring interacts with a thymine methyl group. DZQ complexation is favored at the TGCA site, whereas for MeDZQ the lowest energy complexes were obtained for binding at the TGTC site. The major determinant of the strength of the non-covalent interaction for all the quinone complexes was the angle between the plane of the aromatic ring and the base pair plane. The binding interaction of the quinone forms with an adenine tract was also studied. In this case, the ligand location follows a contour defined by the thymine methyl groups. Both quinones bound less effectively to this sequence than to either the TGCA or TGTC sites. PMID- 8240657 TI - Differentially potentiating effects by dipyridamole on cytotoxicity of 5 fluorouracil against three human maxillary cancer cell lines derived from a single tumor. AB - Dipyridamole (DPM), an inhibitor of nucleoside transport, is a unique potentiator of 5-fluorouracil (5-FU). We examined the combined effects of DPM and 5-FU on three cell lines (IMC-2, IMC-3, IMC-4) derived from a single tumor of a patient with maxillary cancer. DPM at 1.0-2.0 microM potentiated the cytocidal action of 5-FU > 20-fold against IMC-2 cells, while DPM at 10-20 microM potentiated the action of 5-FU against IMC-3 cells by 2-fold and against IMC-4 cells by approximately 4-fold. We examined why DPM differentially potentiated 5-FU against the three cell lines. The three maxillary cell lines showed heterogeneous sensitivities to the combination of 5-FU and DPM. We compared intracellular metabolism of 5-FU in IMC-2, IMC-3 and IMC-4 cells by HPLC. Determination of 5-FU metabolites demonstrated that cellular contents of 5-fluorodeoxyuridine monophosphate (FdUMP) were 104.66, 15.35 and 61.5 fmol/10(6) cells in IMC-2, IMC 3 and IMC-4 cells, respectively, in the presence of 10 microM DPM when the three cell lines had similar levels (0.21-0.33 fmol/10(6) cells) of FdUMP in the absence of DPM. By contrast, cellular levels of 5-FU and 5-fluorouridine triphosphate (FUTP) were not appreciably changed by DPM in three cell lines. The cellular level of thymidylate synthase mRNA in IMC-2 cells was found to be about one-fifth of that in IMC-3 and IMC-4 cells. The differential effects of DPM on the potentiation of 5-FU cytotoxicity might be closely related to the cellular levels of FdUMP and its target enzyme, thymidylate synthase in three human maxillary cancer cell lines. PMID- 8240658 TI - Differentiation: a suitable strategy for cancer chemotherapy? AB - Differentiation therapy focuses on the development and use of specific agents designed to selectively engage the process of terminal differentiation, leading to the eventual elimination of tumorigenic cells and rebalance of normal cellular homeostasis. Extensive in vitro study of the molecular mechanism involved during drug-induced maturation has allowed the realization and application of a differentiation-based therapy to the clinic. Rationalization of this mode of therapy has included the combined use of differentiation agents with low-dose chemotherapy to lessen adverse cytotoxicity and to enhance the efficacy of differentiation agents, allowing some success in their application to conditions resistant to conventional therapy. This review discusses some biological principles that underlie the concept of a differentiation therapy and compares the in vitro and in vivo effectiveness of the two differentiation agents, in particular retinoic acid (RA) and hexamethylene bisacetamide (HMBA). It also evaluates the prospects for differentiation therapy as an effective strategy in the treatment and management of malignancy. PMID- 8240659 TI - Coxsackievirus B4 alters pancreatic glutamate decarboxylase expression in mice soon after infection. AB - The 64,000-M(r) (64K) islet autoantigen, which is considered to be a target protein of beta cell destruction in insulin-dependent diabetes mellitus (IDDM), has recently been identified as the enzyme glutamic acid decarboxylase (GAD). We reported a two- to three-fold increased expression of the antigen in islets of diabetes-susceptible mice following infection with a diabetogenic strain of Coxsackievirus B4 (CB4) at 72-h postinfection (p.i.), a time point of active virus replication in the islets. Most of the infected animals subsequently developed 64K autoantibodies and hyperglycemia. Since the infection increases 64K expression, we have analysed immunoreactive GAD expression with a panel of peptide antisera and two widely-used polyclonal antisera against GAD, and measured GAD activity in the brain, pancreas and islets of these mice. Two isoforms, GAD65 and GAD67, are detected in these tissues from non-infected mice. Both GADs are also present in the infected mice brain at 72 h p.i.; however, their islets contain about three-fold more GAD65, and essentially no detectable GAD67. GAD activity is significantly higher in the brain compared with whole pancreas or islets, and islet GAD activity is higher than pancreas GAD activity. The infection significantly reduces islet GAD activity, but not brain GAD activity. CB4-induced abnormalities in islet GAD expression may play a role in virus-induced diabetes. PMID- 8240660 TI - Expression and functional conservation of the human La (SS-B) nuclear autoantigen in murine cell lines. AB - We have studied the cellular expression and functional conservation of the human La autoantigen across species by introducing genes encoding human La antigen into cultured murine cell lines. In transfected murine fibroblasts and lymphoid cell lines human La was expressed as a predominantly nuclear antigen, with a typical pattern of nuclear speckling. Radiolabeled human La was of the predicted molecular mass (48 kDa) when expressed in murine cells and was associated intracellularly with murine 60 kDa Ro antigen. Under certain conditions of cell lysis the association between human La and murine Ro was disrupted, whereas human La and human Ro complexes remained intact, suggesting the possibility of species specific protein interaction between La/Ro antigens. Expression of human La from the genomic gene construct was associated in some transfectants with the presence of additional high molecular weight bands reactive with anti-La monoclonal antibodies. Cell surface La determinants were not detected on any of the transfected cells. Human La protein associated with many of the same radiolabeled RNAs as the murine La antigen in transfected murine cells, indicating intact RNA binding function of La protein across species. The findings indicate that despite 23% non-identity within the primary structure of human and murine La, the human molecule is functionally highly conserved across species. PMID- 8240661 TI - Inhibition of T-cell proliferation by rat synoviocytes. AB - Antigen presentation by synoviocytes to arthritis-related T-cell clones was studied in Lewis rats. Freshly isolated synoviocytes may be able to take up and process mycobacterial antigens but they seem inefficient as antigen presenting cells. Furthermore, synovial cells inhibited the specific proliferative responses of T lymphocytes by a mechanism which apparently was not antigen-specific or mediated by secreted cytokines. Synovial cells isolated from rats at the period of developing adjuvant arthritis showed a lower inhibitory capacity associated with a lower degree of antigen clearance. These results suggest that synoviocytes might inhibit T-cell responses in normal joints and that this negative control diminishes following arthritis induction. PMID- 8240662 TI - The role of T-helper lymphocytes in priming for experimental autoimmune glomerulonephritis in the BN rat. AB - In our model of experimental autoimmune glomerulonephritis (EAG), BN rats given a single IM injection of homologous glomerular basement membrane (GBM) in FCA develop anti-GBM autoantibodies with focal glomerulonephritis. To investigate the role of lymphocytes in the induction of EAG, we examined the effects of antigen rechallenge and of adoptive cell transfer from donors with nephritis to naive recipients. Groups of animals were rechallenged with GBM in FCA following either low (non-nephritogenic) doses of GBM, or following resolution of EAG. This resulted in an enhanced anti-GBM antibody response in both groups, suggesting the presence of GBM specific T or B memory cells. To investigate this possibility, spleen cells from animals with EAG were transferred into lightly irradiated recipients. There was no significant rise in anti-GBM antibody levels after transfer. However, subsequent challenge with GBM in FCA resulted in an enhanced anti-GBM antibody response by 2 weeks when compared with recipients of normal spleen cells. Cells capable of priming for EAG developed by week 4 after immunization of donors and persisted in the recipients for at least 24 weeks. To investigate which cell type was responsible for this effect, we depleted or positively selected donor spleen cells prior to transfer, using Dynabeads coated with monoclonal antibodies to Th or B lymphocytes. Depletion of Th cells, but not B cells, reduced the enhanced anti-GBM antibody response of recipients challenged with GBM in FCA. Positively selected Th cells, but not B cells, resulted in an enhanced anti-GBM antibody response similar to that in positive controls. These results demonstrate the presence of immunological memory for the autoantigen and show that priming for EAG is mediated by Th lymphocytes. PMID- 8240663 TI - Complementarities and network interactions in AIDS. AB - The contribution of autoimmune phenomena to the development of AIDS is not fully understood. In this paper we provide new evidence that idiotypic network interactions are operative in the development of AIDS. We furthermore present a model that involves two groups of antibodies and other molecules that are related to each other by a network of specific complementarities. We define Group I molecules to include antibodies and other molecules that have direct or indirect similarities with class I MHC molecules, and Group II substances that have similarities with class II MHC. We observe an inverse relationship between Group I antibodies (including anti-Fab) and Group II antibodies (including anti-anti gp120, anti-anti-CD8 and anti-rCD4). The potential roles of members of these two groups in AIDS pathogenesis and/or protection are discussed. PMID- 8240664 TI - Limited T-cell receptor diversity in liver-infiltrating lymphocytes from patients with primary biliary cirrhosis. AB - Primary biliary cirrhosis is associated with the presence of high-titer anti mitochondrial autoantibodies as well as T-cell infiltration of the liver, suggesting the involvement of autoimmune mechanisms. We have studied here the sequences of T-cell receptor alpha and beta chains expressed by T-cell clones derived from liver-infiltrating lymphocytes of two patients with primary biliary cirrhosis. Among the eight clones studied from the first patient, four expressed the same member of the V beta 6 subfamily, associated with either V alpha 4 (three clones) or V alpha 21 (one clone) gene segment. Two other clones expressed an identical V beta 12 transcript, and two in-frame alpha chain transcripts, involving V alpha 2 and V alpha 7 gene segments. From the second patient, eight out of the nine clones were found to rearrange V beta 17-J beta 2.1 and V alpha 3 gene segments. The remaining clone expressed distinct T-cell receptor chains, involving V beta 9 and V alpha 11 gene segments. As deduced from the analysis of their junctional regions, the eight T-cell clones expressing V beta 17/V alpha 3 gene segments derived from only three different T cells. Furthermore, conserved amino acid motifs were found to be encoded in both the alpha and the beta-chain junctional regions. Together, these data show a local amplification of unique T lymphocytes in both patients. The use of identical V beta J beta and V alpha gene segments with similar junctional sequences by three different cells, evidenced in one of the two cases, strengthens the view that liver-infiltrating T lymphocytes are selected locally by autoantigens in PBC. PMID- 8240665 TI - Selection of T lymphocytes in two rheumatoid arthritis patients defines different T-cell receptor V beta repertoires in CD4+ and CD8+ T-cell subsets. AB - To study the selective pressures responsible for the expansion of T cells in rheumatoid arthritis, we constructed cDNA mini-libraries from purified CD4+ and CD8+ T cells prepared from peripheral blood and from synovial fluids of two rheumatoid arthritis patients. Comparison of these libraries by hybridization with specific probes indicated that V beta 2 and V beta 8 transcripts are selectively enriched in the CD4+ synovial fluid lymphocyte population, while V beta 4 was over-represented among both the CD4+ and CD8+ subsets. The enrichment of V beta 14 and V beta 17 observed in synovial fluid T cells of one patient was, however, selectively confined to the CD8+ T-cell subpopulation. Sequence analysis of several V beta 2, V beta 4 and V beta 8 clones, derived from CD4+ cells, revealed a high degree of heterogeneity in the V beta-D beta-J beta junctions, while a more biased utilization of J segments and a more restricted junctional heterogeneity were observed in V beta 4, V beta 14 and V beta 17 clones derived from CD8+ cells. These data suggest that the disease may be induced by the initial activation of a rather heterogeneous population of T-helper cells that are later responsible for the expansion of a more restricted pool of highly specific effector lymphocytes. PMID- 8240666 TI - Monoclonal antibody detection of laminin in minor salivary glands of patients with Sjogren's syndrome. AB - The potential roles of the basement membrane proteins, laminin and fibronectin, and the cytoskeletal protein, tubulin, were assessed in the pathogenesis of Sjogren's Syndrome (SS) by comparing their expressions in SS with normal labial salivary gland (LSG) tissue. Laminin, fibronectin and tubulin expression were determined using well characterized monoclonal antibodies in the peroxidase anti peroxidase technique on formalin-fixed LSG's from patients with SS and normal controls. Characteristic periductal staining for laminin occurred in the LSG's of 14/18 SS patients scored by one observer and 16/18 scored by the second observer. Staining of LSG's for laminin occurred in 2/35 control specimens consisting of 15 normal LSG's and 20 inflammatory lesions with attached normal LSG. The staining which occurred in the two controls was diffuse and 'non-specific' in one case, and indistinguishable from the characteristic periductal staining found in SS in the other case. Among the 20 controls containing inflammatory lesions, four showed diffuse staining for laminin within the actual lesion, but the adjacent LSG's did not stain. No statistically significant difference between SS and normal tissues stained by anti-fibronectin and anti-tubulin was observed. The study concluded that there was an increase in laminin or a laminin-like substance on salivary ductal epithelia of SS patients. This suggests a potential role for laminin in the pathologic mechanism and may indicate that increased laminin expression is a marker for SS. PMID- 8240667 TI - Increased tachykinin receptor gene expression in asthmatic lung and its modulation by steroids. AB - Substance P has several inflammatory effects on the airways mediated via neurokinin 1 receptors (NK1Rs) and, if released from sensory nerves, may amplify the chronic inflammation seen in asthma. Northern blot analysis of NK1R mRNA in lung showed a 52 +/- 10% (S.E.M.; P < 0.01) increase in mRNA in the asthmatic lung compared with non-asthmatic control tissue. NK1R mRNA was reduced by 84.5 +/ 1.9% after incubation with dexamethasone (1 microM) for 3 h (P < 0.01). In contrast, NK2R mRNA was unaltered in asthmatic lungs and dexamethasone treatment had no effect on the level of NK2R mRNA. These results suggest that chronic inflammation in asthma may result in increased NK1R gene expression and that this effect is reversed by glucocorticosteroids. PMID- 8240668 TI - Evolutionary conservation of the insulin gene structure in invertebrates: cloning of the gene encoding molluscan insulin-related peptide III from Lymnaea stagnalis. AB - Although insulins and structurally related peptides are found in vertebrates as well as in invertebrates, it is not clear whether the genes encoding these hormones have emerged from a single ancestral (insulin)-type of gene or, alternatively, have arisen independently through convergent evolution from different types of gene. To investigate this issue, we cloned the gene encoding the molluscan insulin-related peptide III (MIP III) from the freshwater snail, Lymnaea stagnalis. The predicted MIP III preprohormone had the overall organization of preproinsulin, with a signal peptide and A and B chains, connected by two putative C peptides. Although MIP III was found to share key features with vertebrate insulins, it also had unique structural characteristics in common with the previously identified MIPs I and II, thus forming a distinct class of MIP peptides within the insulin superfamily. MIP III is synthesized in neurones in the brain. It is encoded by a gene with the overall organization of the vertebrate insulin genes, with three exons and two introns, of which the second intron interrupts the coding region of the C peptides. Our data therefore demonstrate that in the Archaemetazoa, the common ancestor of the vertebrates and invertebrates, a primordial peptide with a two-chain insulin configuration encoded by a primordial insulin-type gene must have been present. PMID- 8240669 TI - Endotoxin-stimulated production of rat hypothalamic interleukin-1 beta in vivo and in vitro, measured by specific immunoradiometric assay. AB - Regulation of a number of aspects of the acute-phase response, including induction of fever and activation of the hypothalamo-pituitary-adrenal axis, occurs within the hypothalamus. The acute-phase response appears to be co ordinated by the inflammatory cytokine interleukin-1 (IL-1). A number of studies using hybridization techniques to measure IL-1 gene expression and immunocytochemistry to localize immunoactive IL-1 have established the concept that the central nervous system, and in particular the hypothalamus, is a site of IL-1 production, and that levels increase in response to inflammatory stimuli. In this report we present data on the levels of IL-1 beta produced in the rat hypothalamus using quantitative immunoassay techniques. Bacterial endotoxin, administered to rats in vivo, evoked increases in hypothalamic IL-1 beta levels which were significant within 1 h, and reached maximum levels at 5-10 h. The response to endotoxin was dose-related, and levels reached in hypothalamic extracts corresponded to intrahypothalamic levels of the order of 20 ng/ml. During short-term in-vitro culture of rat hypothalami, endotoxin stimulated a dose-related increase in both the synthesis and the secretion of IL-1 beta, which reached similar levels to those seen after in-vivo stimulation. Hypothalami obtained from animals stimulated with endotoxin in vivo did not, however, show any evidence of persistent stimulation of IL-1 beta production when subsequently cultured in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240670 TI - The peroxisome proliferator-activated receptor:retinoid X receptor heterodimer is activated by fatty acids and fibrate hypolipidaemic drugs. AB - The peroxisome proliferator-activated receptor (PPAR) is a member of the steroid hormone receptor superfamily and is activated by a variety of fibrate hypolipidaemic drugs and non-genotoxic rodent hepatocarcinogens that are collectively termed peroxisome proliferators. A key marker of peroxisome proliferator action is the peroxisomal enzyme acyl CoA oxidase, which is elevated about tenfold in the livers of treated rodents. We have previously shown that a peroxisome proliferator response element (PPRE) is located 570 bp upstream of the rat peroxisomal acyl CoA oxidase gene and that PPAR binds to it. We show here that the retinoid X receptor (RXR) is required for PPAR to bind to the PPRE, and that the RXR ligand, 9-cis retinoic acid, enhances PPAR action. Retinoids may therefore modulate the action of peroxisome proliferators and PPAR may interfere with retinoid action, perhaps providing one mechanism to explain the toxicity of peroxisome proliferators. We have also shown that a variety of hypolipidaemic drugs and fatty acids can activate PPAR. This supports the suggestion that the physiological role of PPAR is to regulate fatty acid homeostasis, and provides further evidence that PPAR is the target of the fibrate class of hypolipidaemic drugs. Finally, we have demonstrated that a metabolically stabilized fatty acid is a potent PPAR activator, suggesting that fatty acids, or their acyl CoA derivatives, may be the natural ligands of PPAR. PMID- 8240671 TI - Construction of an ecotropic retroviral vector expressing human insulin-like growth factor-I. AB - There is increasing evidence that IGF-I plays an autocrine role in a wide range of human tumours, including, in particular, adenomas of the thyroid epithelium. To investigate this further, we set out to generate a retrovirus vector which would permit experimental manipulation of the expression of IGF-I in normal and neoplastic epithelial cells. We describe here the construction and validation of a high-titre ecotropic vector which transduces stable expression and secretion of human IGF-IA, as shown by analysis of mRNA and conditioned medium from rodent epithelial target cells. This vector should be a useful tool for assessing the contribution of abnormal IGF-I expression to the neoplastic phenotype. PMID- 8240672 TI - Hormonal study of a human mixed follicular and medullary thyroid carcinoma. AB - We studied the hormonal secretion of a human mixed follicular and medullary carcinoma. Thyroglobulin (Tg) secretion, especially by large cells and sometimes by small ones, was visualized with immunoenzymatic staining. Calcitonin (CT) was produced by small spindle-shaped cells. Moreover, immunofluorescence double staining performed on the resected thyroid tissue showed the secretion of both Tg and CT in a small number of cells. The cells lost their hormonal secretion after 2 months of culture. Hormonal secretion was modulated by different additives in the medium. Tg secretion was induced when TSH was added to the culture medium; the maximal effect was produced with the addition of 1 mU TSH/ml and 1 microM cortisol, which potentiated the effect of TSH on Tg production. A durable Tg secretion was obtained by embedding the cells in Engelbretch-Hohn-Swarn (EHS) tumour matrix. The CT production was reinduced by the addition of 4 mM Ca2+, 1 microM glucagon and 1 microM cortisol to the culture medium. These findings show that different cells are found in a mixed follicular and medullary carcinoma, some of which can secrete both CT and Tg. They can remain differentiated for a long period after being embedded in EHS tumour matrix with Ca2+ and hormonal components. PMID- 8240673 TI - Angiotensin II receptor isoforms in the rat adrenal gland: studies with the selective subtype antagonists DuP 753 and CGP42112A. AB - The angiotensin II (Ang II)-binding sites in rat adrenal gland membranes were characterized using 125I-radiolabelled Ang II. While Scatchard analysis identified a single population of Ang II receptor sites, isoelectric focusing (IEF) on polyacrylamide gels revealed four peaks of specific Ang II binding which migrated to isoelectric points (pI values) 6.8, 6.7, 6.5 and 6.3. In binding assays in the presence of an excess of the Ang II receptor AT1 subtype antagonist DuP 753, a monophasic dose-dependent displacement of 125I-labelled Ang II binding by the Ang II receptor AT2 subtype antagonist CGP42112A was observed, and vice versa. In this system, reduction of disulphide bridges using 1 mmol dithiothreitol (DTT)/l markedly increased the number of binding sites in the adrenal zona glomerulosa without affecting receptor affinity. Using IEF, it was found that both DuP 753 and CGP42112A were able to reduce specific binding of each of the four peaks to some extent. However, the predominant effect of DuP 753 was to reduce the labelling of the isoform at pI 6.7 substantially, while CGP42112A significantly inhibited the specific 125I-labelled Ang II binding to the pI 6.3 isoform. When DuP 753 and CGP42112A were used together, specific binding of 125I-labelled Ang II to the isoforms of pI values 6.8, 6.7 and 6.3 was completely eliminated. These data suggest that the four peaks of specific binding found may be composed of different isoforms of both AT1 and AT2 receptor subtypes and that the Ang II receptor isoforms which migrated to pI 6.7 and pI 6.3 are predominantly composed of AT1 and AT2 receptor subtypes respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240674 TI - Cytochrome P-450 17 alpha-hydroxylase protein and mRNA in the testis of the testicular feminized (Tfm) mouse. AB - The testicular feminized (Tfm) mouse lacks functional androgen receptors and develops with a female external phenotype and internal testes. The testes of these animals contain normal, or close to normal, numbers of Leydig cells but secrete very low amounts of androgen due to a lack of 17 alpha-hydroxylase activity. To determine whether this loss of activity is due to a lack of enzyme synthesis or a change in catalytic activity we have examined 17 alpha-hydroxylase cytochrome P-450 (P-450(17 alpha)) protein and mRNA levels in the testes of Tfm mice. Levels of P-450(17 alpha) protein were measured by immunoblotting, while mRNA was measured following reverse transcription (RT) and amplification by the polymerase chain reaction (PCR). Conditions for RT-PCR were determined which allowed semiquantification of P-450(17 alpha) mRNA relative to beta-actin mRNA. In extracts of Tfm testes P-450(17 alpha) protein was undetectable using antiserum against porcine P-450(17 alpha). In contrast, a protein of around 54 kDa was clearly detectable in extracts of control cryptorchid testes. Using RT PCR, P-450(17 alpha) mRNA was detectable in both control and Tfm testes but, expressed in terms of beta-actin mRNA, levels of P-450(17 alpha) mRNA in control testes were 40-fold higher than those in Tfm testes. If the total amount of RNA extracted from each testis is taken into account then P-450(17 alpha) mRNA levels per testis were up to 400-fold higher in control testes. These results show that the reduced level of 17 alpha-hydroxylase activity in Tfm testes is related to reduced protein synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240675 TI - The nature and significance of multiple isoforms of the oestrogen receptor in breast tumours. AB - Oestrogen receptors (ERs) in breast tumours are highly heterogeneous. In previous studies we have shown that at least four isoforms may exist. These migrate in isoelectric focusing (IEF) gels to isoelectric points (pI values) 6.1, 6.3, 6.6 and 6.8. Of these the first (pI 6.1) corresponds to the 8S isoform as detected by sucrose gradient fractionation, while the others all sediment at 4S. In a series of 66 breast tumours it was found that those at pI 6.3 and pI 6.8 were significantly correlated with the presence of progesterone receptors. To characterize the isoforms more fully, ER isoforms labelled by [3H]oestradiol binding were fractionated by IEF. The results were compared with those obtained after sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using the H222 anti-ER monoclonal antibody. In other experiments, tumour ER isoforms were covalently labelled with [ring-3H] tamoxifen aziridine and separated by IEF. The individual isoforms were electroeluted from the IEF gel and further analysed by SDS-PAGE and non-denaturing PAGE. In summary, the evidence shows that the isoforms of pI values 6.3, 6.8 and 6.6 have molecular masses of 50, 65 and 70 kDa respectively. In addition, all three of these isoforms, i.e. the pI 6.3, 6.8 and 6.6 isoforms, could form dimers. We conclude that the three isoforms sedimenting at 4S have the capacity to form dimers and thus may have the potential for binding to oestrogen response elements in the genome. PMID- 8240676 TI - Characterization of rabbit kappa-casein cDNA: control of kappa-casein gene expression in vivo and in vitro. AB - The rabbit kappa-casein cDNA was cloned and sequenced. One of the isolated clones included almost the entire 5' end, while another clone corresponded to the 3' end of the cDNA. No polyadenylation site was found and therefore this clone did not harbour the complete cDNA. The amino acid sequence of a full-length protein was deduced from the nucleotide sequence obtained for this partial cDNA. It revealed the presence of a chymosin cleavage site and five potential phosphorylation sites. Rabbit kappa-casein was compared with those already described in other species. The rabbit sequence is closer to the ovine than to the mouse sequence. This result supports the idea that Lagomorpha are not closer to Rodentia than to Artiodactyla. The cDNA described above was used to study kappa-casein gene expression in the rabbit mammary gland. This expression was induced primarily by prolactin in mammary gland organoids and was similar to alpha s1-casein gene expression in vivo. The kappa-casein gene present in the casein gene locus is thus subject to the same regulation as the alpha s1-casein gene, although it has evolved from a fibrinogen gene. PMID- 8240677 TI - [Quantitative bibliographic review on the use of anabolic hormones with steroidogenic action in ruminants for meat production. II. Principal mode of action]. AB - The hypotheses on the modes of action of hormonal anabolic agents in growing animals have been reviewed in more than 120 recent publications. The mechanisms of action are still not fully understood. Androgens such as testosterone and estrogens such as oestradiol-17 beta (E-17 beta) may act in different ways: firstly, testosterone (and probably also E-17 beta) acts directly on different tissues, and particularly at the level of the muscle cell by binding to a specific receptor. The hormone-receptor complex interacts with the nuclear receptor located in the chromatin and enhances protein synthesis (and probably also protein degradation). Trenbolone acetate (TBA) reduces protein synthesis and to a greater extent protein degradation. This action of TBA could take place via a reduction in the activity of catabolic glucocorticoids, either by a diminution in their secretion, or by displacing them from their receptor, or by reducing the number of receptors. Secondly, an indirect action of anabolic hormones is probable via the modifications in activity of other growth-regulating hormones. Growth hormone and insulin-like growth factor-I concentrations are enhanced by E 17 beta, diethylstilbestrol, zeranol and testosterone but not by TBA. Insulin appears to be indirectly enhanced by estrogens through an increase in growth hormone, whereas androgens reduce insulin levels. Thyroid hormone (tri- and tetra iodothyronine) activity is reduced by androgens, whereas the action of oestrogens depends on the physiological maturity of the animal. The modes of action of these anabolic hormones are discussed in relation to growth rate and body composition. PMID- 8240678 TI - [Nutritive value of white lupin (Lupinus albus var Lutop) in rainbow trout (Oncorhynchus mykiss). Effect of extrusion cooking]. AB - Two experiments were conducted in rainbow trout (Oncorhynchus mykiss) to assess the nutritional value of diets containing crude (LC) or extruded at 120 degrees C (LE1) and 145 degrees C (LE2) lupin var "Lutop" as a partial protein source. In the first experiment, the apparent digestibility coefficient (ADC) of diets with 20 and 30% lupin was measured. In a second trial, the effects of diet containing 20% crude and extruded lupin on growth performance of trout were analyzed. It was found that the ADC of dry matter, proteins and lipids decreased with the incorporation of 20% crude lupin. The extrusion-cooking process increased the digestive utilization of nutrients, especially the nitrogen-free extract. After 83 d of feeding, decreased growth performance was observed in trout fed on LC diet. Those fed diets LE1 or LE2 showed increased growth, food intake, food efficiency and protein retention, attributed to the disappearance of antinutritional factors in extruded lupin. PMID- 8240679 TI - Energy resources of spermatozoa of the rainbow trout Oncorhynchus mykiss (Pisces, Teleostei). AB - Spermatozoa of Oncorhynchus mykiss have the enzymatic capacity for glycolysis, for triglyceride and phospholipid catabolism and triglyceride synthesis. They lack glucosidase activity and are therefore not able to utilize polysaccharides as energy resources. In motile spermatozoa glycolysis occurs during the first 30 s of motility and--when motility is initiated in a physiological saline solution- triglyceride catabolism is used for the regeneration of ATP levels after motility has ended. When immotile spermatozoa are incubated with the seminal plasma or in physiological saline solution they behave similarly as regards utilization of their primary energy reserves: glycolysis as well as catabolism of triglycerides occurs. In approximately 60% of the semen samples, spermatozoa synthesize triglycerides at the onset of incubation. Possible physiological reasons for triglyceride synthesis have been discussed. PMID- 8240680 TI - Plasma levels of growth hormone and insulin-like growth factor-I and -II from 2 to 6 weeks of age in meat-type chickens selected for 6-week body weight or for feed conversion and reared under high or normal environmental temperature conditions. AB - The aim of this study was to compare the effect of high (33 degrees C) and normal (33-20 degrees C) rearing temperature on growth and plasma levels of the somatotrophic hormones of 2 genetic lines of broiler chickens selected for 6-wk body weight (GL-line) or for feed conversion between 3 and 6 wk of age (FC-line) or for feed conversion between 3 and 6 wk of age (FC-line). Blood samples were collected weekly and analysed for growth hormone (GH) and insulin-like growth factors (IGF)-I and -II levels by RIA. The growth-depressing effect of the HT treatment was more pronounced in the heavier GL-line and in males. A similar age related pattern for all hormones studied was observed with the highest levels between 2 and 4 wk of age. FC-line chickens and males had consistently higher plasma GH levels than GL-line chickens and females respectively. No consistent effect of rearing temperature on plasma GH levels were observed. At 2 wk of age, HT-treatment resulted in higher plasma IGF-I levels while this was reversed from 3 wk of age onwards. GL-line chickens had significantly higher plasma IGF-I levels at 2, 3 and 4 wk of age. No consistent effect of sex on plasma IGF-I levels could be observed. For the whole period studied, GL-line chickens had significantly higher plasma IGF-II levels than FC-line chickens. No consistent effect of sex or temperature treatment on plasma IGF-II levels was observed. PMID- 8240681 TI - Exogenous and endogenous contributions to nitrogen fluxes in the digestive tract of pigs fed a casein diet. III. Recycling of endogenous nitrogen. AB - The aim of the present study was to measure the incorporation of infused 15N in blood fractions, urine, digesta, faeces and in the exocrine pancreatic and biliary secretions, in order to estimate the endogenous part of nitrogen in the ileal digesta and in the faeces of pigs fed a casein diet and to calculate the total endogenous nitrogen secretion as well as its recycling in the digestive tract. For 8 d 11 Large White female pigs (50.1 +/- 1.8 kg) received a continuous infusion of L-[15N]leucine via a catheter in the jugular vein. The 15N-enrichment was measured in several fractions. The 15N-level of the pancreatic juice was higher than that in the biliary secretion, TCA-blood fractions, and urine during the whole experimental period. Using the 15N-isotope dilution method it was found that casein was completely digested up to the terminal ileum and that all the nitrogen in the ileal digesta was of endogenous origin. The total endogenous secretion was estimated at approximately 11 g N/d. The reabsorption of endogenous nitrogen amounted to 79% up to the end of the small intestine and 88% over the whole digestive tract. PMID- 8240682 TI - [Influence of the number of corpora lutea on the release of luteal oxytocin, the transfer of alveolar milk to the cistern and milk production in the ewe]. AB - This experiment was conducted in 59 Lacaune breed ewes in order to compare milk production and milk distribution between alveolar and cisternal storage after superovulation. After a corpora lutea (CL)-free control period, the ewes were superovulated by different treatments (experimental period) and 5 classes were differentiated according to the number of corpora lutea observed (0, 1, 2, 3 to 6 and > 6 CL respectively (group A (n = 20), B (n = 7), C (n = 14), D (n = 7), E (n = 11)). Our results showed a positive correlation between the number of corpora lutea and the oxytocin and progesterone levels in plasma, total milk production and cisternal volume, and a negative correlation with alveolar volume. The milk production at the evening milking for groups A and E (388.6 and 384.8 ml respectively during the control period) respectively reached 321.7 ml (-17.2%) and 413.4 ml (+7.4%) during the experimental period; ie, a 24.6% difference between these 2 extreme groups. These results could likely be explained by oxytocin levels reaching those obtained during milking and by the effect of milk transfer from the alveolar to the cisternal lumen. Additionally, progesterone could act directly on milk synthesis at the level of the secretory cells. PMID- 8240683 TI - Effects of body composition variations on the duration of the postpartum anovulatory period in milked ewes submitted to two different photoperiods. AB - Forty-four dairy ewes were placed at lambing under long (n = 22) or short (n = 22) artificial photoperiods. In each photoperiodic treatment 2 diets (high or low) were offered to 11 group-fed ewes. The duration of the anovulatory period (DAP, in d) was estimated by bi-weekly blood progesterone measurements. Within 2 months, ewes exposed to the long photoperiod had no resumption of ovulation, while most of the ewes (18/22) exposed to the short photoperiod ovulated. In vivo measurements of body composition (D2O) showed that the shortest DAP (ie 36 d) was observed for ewes which gained 1 kg body lipids during the first month postpartum. Above and below this threshold, DAP increased. Other relationships between DAP and ewe body composition variations (dynamic) and/or total chemical body mass (static) were determined. In dairy ewes, it seems that the dynamic aspects are more closely related to DAP than the static aspects. Results were compared to the available literature on anestrus in dairy cows. PMID- 8240684 TI - Fetal blood natural killer cells in human pregnancy. AB - Flow cytometry was used to determine the number of circulating natural killer (NK) cells in 170 pregnancies undergoing cordocentesis for prenatal diagnosis at 17-38 weeks gestation and 16 undergoing cardiocentesis before social termination of pregnancy at 13-17 weeks. The number of NK cells decreased exponentially with gestation from a mean of 0.26 x 10(9)/l at 13 weeks to 0.09 x 10(9)/l at 38 weeks (r = -0.356, p < 0.001). When expressed as a percentage of the total number of lymphocytes, the proportion of NK cells decreased from a mean of 29% at 13 weeks to 6% at 38 weeks (r = 0.680, p < 0.0001). These data suggest that in early pregnancy the fetal immune response is dependent on innate rather than adaptive mechanisms. PMID- 8240685 TI - Immune thrombocytopenic purpura and percutaneous umbilical blood sampling: an open question. AB - We performed a retrospective study of 26 pregnancies with chronic immune thrombocytopenic purpura (ITP) or incidental ITP. Thirteen pregnancies were followed without the use of percutaneous umbilical blood sampling (PUBS) and 13 were followed sampling PUBS at 36-40 weeks of gestation. The overall prevalence of neonatal thrombocytopenia was 27%: 50% in chronic ITP, 12.5% in incidental ITP. Symptomatic thrombocytopenia occurred only in infants born to mothers with chronic ITP. Without the use of PUBS, 2 symptomatic thrombocytopenic fetuses were vaginally delivered and there was a high rate of cesarean sections in normal fetuses. Owing to PUBS, a decrease of the cesarean section rate in normal fetuses was observed, but one fetal bradycardia (due to the technique) was encountered. PUBS resulted helpful to indicate the best route of delivery and reducing unnecessary cesarean sections in women with chronic ITP, but in presence of incidental ITP the use of PUBS remains an open question. PMID- 8240686 TI - Fetal and neonatal hematologic parameters in red cell alloimmunization: predicting the need for late neonatal transfusions. AB - The purpose of this study was to determine whether the need for late neonatal transfusion can be predicted in fetuses who received intrauterine transfusions. The records of 36 patients who underwent intrauterine transfusion were reviewed. Newborns who required late transfusions had a lower reticulocyte count at their last intrauterine transfusion, a longer duration between their lowest fetal reticulocyte count and delivery, and a higher umbilical cord blood hemoglobin and percent adult red cells on Kleihauer-Betke stain at delivery. The need for late transfusions appears to be related to the extent and duration of fetal bone marrow suppression. PMID- 8240687 TI - Amniotic fluid platelet factor 4 and beta-thromboglobulin. AB - Platelet activating factor (PAF), a powerful platelet activator, has been identified in human embryos and fetuses, and may induce fetal lung maturation. The potential effect of PAF on fetal platelets as indicated by release of beta thromboglobulin (BTG) and platelet factor 4 (PF4) has not been investigated. We measured BTG and PF4 in amniotic fluid from 78 genetic and 35 pulmonary maturity amniocenteses. BTG and PF4 were higher in the genetic amniocentesis samples (p < 0.001 in each case) than in the lung maturity samples. BTG and PF4 did not correlate with the pulmonary maturity parameters as measured by the lecithin to sphingomyelin ratio and phosphatidylglycerol concentration. Our findings suggest a fetal origin of BTG and PF4 in amniotic fluid. PMID- 8240688 TI - Amniotic fluid acetylcholinesterase is found in gastroschisis but not omphalocele. AB - Amniotic fluid acetylcholinesterase (ACHE) has been used to evaluate neutral tube defects. It has also been detected in ventral wall defects. However, the role of ACHE to differentiate between omphalocele and gastroschisis has not been established. We examined amniotic fluid ACHE in 16 pregnancies complicated by gastroschisis and 8 by omphalocele. One ruptured omphalocele was excluded. ACHE was negative in all 7 omphaloceles and either positive or suspicious in all gastroschises (chi 2 = 17.3, p < 0.0001). Amniotic fluid ACHE may be useful to differentiate between omphalocele and gastroschisis. PMID- 8240689 TI - Changing dysmorphology of trisomy 18 during midtrimester. AB - A fetus affected by trisomy 18 was observed serially on ultrasound from 14 to 21 weeks of gestation. The earliest and sole dysmorphic feature noted at 14 weeks of gestation were nonseptated nuchal cysts. At 17 weeks the ultrasound examination was considered normal. However, at 21 weeks gestation, bilateral choroid plexus cysts, clubfoot, growth retardation and overlapping fingers were observed. These findings suggest that the dysmorphology associated with trisomy 18 is dynamic in nature and may change along the course of pregnancy. PMID- 8240690 TI - Age of gestation (size) at embryonic demise: tailoring counseling for lethal versus potentially viable aneuploidy. AB - The incidence of chromosomal abnormalities in spontaneous abortions is much higher than commonly appreciated. Using postmortem chorionic villus sampling (as we have previously described), our much increased yield of results allows us to divide embryos into two groups based upon potential viability. Embryonic size at the time of demise correlated with viability, i.e., the more 'viable' the embryo for term survival. Even in the absence of chromosomal results, the size of the embryonic pole can be used to infer, to a degree, the type of abnormality. PMID- 8240691 TI - Prenatal diagnosis of tetraploidy. AB - A pregnancy was terminated at 24 weeks of amenorrhea when tetraploidy (92 XXXX) was diagnosed in fetal blood subsequent to ultrasonographic detection of a polymalformation syndrome. The severity of the neurological deficit in tetraploid infants and their death before 2 years of age require that prenatal diagnosis by cordocentesis be performed for analysis of fetal blood in cases of equivocal and nonspecific polymalformation syndrome and justify that medically-induced termination of pregnancy is suggested in the event of intrauterine tetraploidy diagnosis. PMID- 8240692 TI - Combined effect of terbutaline and betamethasone on glucose homeostasis in preterm labor. AB - The short- and long-term effects of simultaneous administration of terbutaline and betamethasone were investigated in 8 gravidas treated for preterm labor. Their plasma concentrations of glucose, insulin, glucagon, C-peptide, lactate and potassium were compared to a control group receiving intravenous magnesium sulfate and betamethasone. The patients on terbutaline therapy had a marked hyperglycemia at 11 h which remained elevated for 48 h. There was a simultaneous rise in plasma insulin and C-peptide, and a fall in plasma glucagon. Lactate levels were markedly elevated. Only 1 of the 8 patients had an abnormal glucose tolerance test at 1 week of therapy. The metabolic changes of control patients were minimal in comparison and there was no lacticacidemia. This suggests that glucocorticoids potentiate the hyperglycemic response of terbutaline. PMID- 8240693 TI - Fetal ovarian cysts: diagnostic and therapeutic role for intrauterine aspiration. AB - Antenatal diagnoses of fetal ovarian cysts have not usually been confirmed until postnatal surgery. We describe 2 cases of hemorrhage into fetal ovarian cysts in which cyst aspiration in utero allowed both confirmation of the diagnosis prenatally and obviated the need for neonatal surgery. In both cases, cytology of the cyst aspirate demonstrated luteinized granulosa cells and biochemistry showed estradiol levels of > 10,000 pmol/l, indicating ovarian etiology. Hemorrhage, which had been suspected on ultrasound, was confirmed by cytology, showing hemosiderin-laden macrophages. There was no evidence of recurrence in either case following aspiration. The described association of fetal hypothyroidism was excluded by testing thyroid function in cord blood and/or cyst aspirate. We suggest that intrauterine aspiration contributes to the management of fetal ovarian cysts by confirming their ovarian origin, demonstrating the presence or absence of hemorrhage and facilitating preservation of ovarian function both by reducing the risk of torsion and the need for neonatal surgery. PMID- 8240694 TI - Unusual fetal movement in congenital myotonic dystrophy. AB - Congenital myotonic dystrophy is the most extreme and early expression of a multisystem autosomal dominant disorder. Decreased fetal and infantile movement is a defining feature. We report a case of congenital myotonic dystrophy with an unusual pattern of persistent repetitive fetal movements detected by a Doppler fetal movement detector. PMID- 8240695 TI - Immediate delivery room repair of fetal abdominal wall defects. AB - Eighteen patients with a prenatal diagnosis of fetal abdominal wall defect were delivered by cesarean section and repaired either immediately (begun within 15 min, n = 9), or by the traditional (delayed) method (n = 9, average delay = 4.4 h). Neonates repaired immediately had comparable gestational ages and birthweight, however, subjectively had less edematous bowel with less fibrous peel. These fetuses were more likely to be closed primarily (7/9 vs. 4/9), spent less time on a ventilator (8.1 vs. 17.9 days), seemed to be fed sooner (7.6 vs. 17.9 days), and discharged home earlier (14.3 vs. 24.0 days). Our results suggest that for fetuses delivered by cesarean section, early defect repair may reduce bowel edema and fibrous peel formation thus facilitating primary closure, with earlier ventilator weaning, feeding and discharge home. PMID- 8240696 TI - White noise does not induce fetal sleep. AB - White noise has been shown to induce sleep in newborns. We sought to examine whether this type of sound will also induce a quiet state in the fetus. Twenty two fetuses at 36-41 weeks of gestation were exposed to white noise during an active state. The sound was delivered for 5 min at an intensity of 100 dB. No significant change in fetal activity was noted following the sound. PMID- 8240697 TI - A case of fetal intestinal tract distensions: prenatal biochemical and ultrasound evaluation. AB - Fetal intestinal tract abnormalities can be pregnancy complications at all maternal ages. There have been reports of ultrasound identification of these abnormalities, particularly of fetal intestinal obstructions. We report the first case to our knowledge of elevated levels of the intestinal isozyme, alkaline phosphatase, in the amniotic fluid and blood serum and also the presence of prealbumin in the amniotic fluid of a fetus with extensive intestinal distension and polyhydramnios. It was revealed that the alkaline phosphatase intestinal isozyme as well as protein electrophoresis for abnormal albumin bands in the amniotic fluid are valuable markers for the early diagnosis of the above mentioned fetal abnormalities. PMID- 8240698 TI - Multifetal pregnancy reduction: is it always justified? AB - In the present study we summarized the world literature on first trimester multifetal pregnancy reduction between 1985 and 1992 and added our own experience. Our aim was to reach a comprehensive view on the survival rate of reduced high multifetal pregnancies, as the data in various studies are somewhat confusing, possibly as a result of the small number of cases. The data so gathered were classified by the number of fetuses. Of 94 triplet pregnancies reduced to twins 91.6% survived. Of 108 quadruplet pregnancies reduced to twins 92.6% survived. Of 40 quintuplet pregnancies 72.5% survived, and of 93 septuplets or more 87.1% survived. The outcome of pregnancies reduced to triplets did not differ from the outcome of pregnancies reduced to twins, with a survival rate of 85.7 and 72.5%, respectively. In our opinion, in view of the infertility history of many of these patients, the risk of pregnancy loss, the high survival rate of triplets and fetal reduction in triplet pregnancies in this group of patients may be unjustified. Accordingly, our policy in cases where the patients demand or when reduction is done early in pregnancy to reduce the triplets. PMID- 8240699 TI - [Management of infantile diarrhea by physicians in outpatient services in Peru]. AB - This study investigated the practices followed by physicians in the clinical, therapeutic, and dietary management of children under the age of 5 with diarrhea. A total of 148 physicians were observed while they provided outpatient services at various public and private health institutions in two areas of Peru. They were also interviewed to assess their theoretical knowledge about the proper management of diarrhea in children. The physicians treated 222 children under 5 with diarrhea. It was observed that in taking their clinical histories the physicians ascertained the duration of the problem, the number of bowel movements, their consistency, and the presence of fecal mucus or blood in 175 (79%), 158 (71%), 140 (63%), and 153 (69%) cases, respectively. Antibiotics were prescribed for 130 children (58%), and 102 (46%) were not given any oral rehydration therapy (ORT). The oral rehydration salts (ORS) were recommended to only 40 (18%) of the children. On the other hand, during the interviews only 38 physicians (26%) indicated that they prescribe drugs for diarrhea, while 110 (74%) said that they recommend ORT. Fewer than half the mothers received recommendations regarding the type of food that they should give their children during the diarrhea episode. The results reveal a sizable discrepancy between theory and practice in the medical management of childhood diarrhea, which means that it is necessary to instill in physicians the importance of assessing and treating children with diarrhea according to a uniform and systematic scheme that includes careful therapeutic and dietary guidance. PMID- 8240700 TI - [Human papilloma virus in the etiology of cervicouterine cancer]. AB - The objectives of this study were to confirm the hypothesis that invasive carcinoma of the uterine cervix and the precursors of that condition are most often caused by human papilloma virus (HPV) infections, and to determine whether or not other risk factors are involved in the neoplastic process. For this purpose, four concurrent case-control studies were carried out. Two included cases of invasive cervical cancer and population-based controls. The other two included cases of in situ carcinoma and controls. Research was carried out in nine provinces of Spain and in Cali, Colombia. The identification of cases took place between June 1985 and June 1988. The studies included 436 incident cases of invasive carcinoma and 387 controls, selected at random from the corresponding populations, and 525 cases of in situ carcinoma and 512 controls paired by age, place of recruitment, and date that cytological specimens were taken from the women participating in cytological screening programs. Exposure to HPV was detected through hybridization tests after amplification by polymerase chain reaction (PCR) in exfoliated cervical cells from cases and controls. Exposure to HPV was the principal risk factor in the four studies. For invasive carcinoma, the relative risk and 95% confidence interval were 46.2 (18.5-115.1) in Spain and 15.6 (6.9-34.7) in Colombia. For in situ carcinoma, the figures were 56.9 (24.8 130.6) in Spain and 15.5 (8.2-29.4) in Colombia. This strong association was specific for types 16, 18, 31, 33, and 35 as well as still-unclassified HPV types.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240701 TI - [Stability of oral rehydration solutions in various packing materials]. AB - The aim of this research was to provide data to select an ideal packing material to preserve the integrity of oral rehydration salts (ORS), a therapeutic formulation that is essential in developing countries. Previous research had verified that water is the factor that most affects ORS stability. For this research, a pharmaceutical industry prepared an ORS batch that was packed in six different types of packing materials. The humidity determination was made after storage of samples during 36 weeks at room temperature, at room temperature with 76% relative humidity, and at 40 degrees C with 80% relative humidity. The moisture in the samples was measured at pre-determined intervals using methods like loss on drying at 50 degrees C and the Karl Fischer method. The results indicated that the most efficient packing material was composed of 18g polyester, 35 g aluminum, and 50 g polyethylene (packing material no. 6). PMID- 8240702 TI - [Medical equipment companies and their ties with technology development centers in Mexico]. AB - The purpose of this study was to determine the characteristics of the companies that produce, distribute, and service medical equipment in Mexico and the factors related to whether or not they had established ties with research and technology development centers. The data analyzed came from a survey of such companies carried out in Mexico City and environs in 1989. The information was updated in 1991. Multivariate analyses were carried out in order to identify the characteristics of companies that had established ties or wished to do so and the areas of interest of those companies. Of 208 companies surveyed, only 23% had ties with research centers. The companies that had such ties or were interested in establishing them tended to invest in research and to have made plans for expansion. The establishment of ties appeared to be a two-way process, with positive consequences for the companies involved, the research centers, and the health sector. It was concluded that it would be advantageous to design programs to promote ties with companies having the characteristics mentioned. PMID- 8240703 TI - von Willebrand's disease. Pathogenesis and clinical aspects. PMID- 8240704 TI - Clinical significance of the estrogen regulated pS2 protein in mammary tumors. AB - A third of breast cancers are estrogen dependent and respond to endocrine therapy. The estrogen receptor (ER) was the first marker used to predict the responses to treatment, and two-thirds of ER positive tumors show a favourable response. Several estrogen-regulated proteins were further studied in a search to enhance the prediction accuracy of ER status: progesterone receptors, 24-K heat shock protein, cathepsin D, and recently pS2 protein. The pS2 gene, also named BCEI, pNR-2 [4], Md2, was first identified by two groups using differential screening of a complementary DNA library derived from a human breast carcinoma cell line (MCF-7) grown with and without estrogens. Later on two independent English groups and a Japanese group identified a gene similar to pS2. The pS2 mRNA, relatively abundant (0.8%) in the MCF-7 cell line when stimulated by estrogens, encodes a cystein-rich, 84 aminoacids peptide which is secreted by breast cancer cells. The expression of the pS2 gene, pS2 protein assays in tumor cytosols and more recently pS2 detection by immunocytochemistry, have been described in several series of breast cancers. PMID- 8240705 TI - Anticoagulant therapy with ancrod. PMID- 8240706 TI - Radiation and chemotherapy injury: pathophysiology, diagnosis, and treatment. AB - The text in general is not meant to represent the participants' entire presentations. The lecture presenters in general are not responsible for the summaries, and cannot necessarily be assumed to agree with all that is stated, but they deserve credit for providing the lecture and handout material on which the summaries are based, and in most cases have contributed far more to the summaries than I have. PMID- 8240707 TI - Role of the dorsal prestriate cortex in visuospatial configural discrimination by monkeys. AB - Cynomolgus monkeys (Macaca fascicularis) learned a series of visuospatial configural discriminations in which particular discriminative stimulus objects were rewarded only in particular spatial locations. For example, object X was rewarded if it was on the left but not if it was on the right. After ablation of part of the dorsal prestriate cortex they were impaired in learning discriminations of this kind. The same animals were not impaired in learning visual object discriminations in which spatial position was irrelevant, nor in learning spatial discriminations in which object identity was irrelevant. The results were compared with previously reported results from fornix transection in the same tasks; the deficit following dorsal prestriate ablation in visuospatial configural discrimination learning was similar in severity to that which followed fornix transection. The results show that the dorsal prestriate area has a more general role in visuospatial processing than was known hitherto, and they suggest that it interacts with the hippocampal formation and fornix in visuospatial memory tasks. PMID- 8240708 TI - The parabrachial region as a possible region modulating simultaneously pain and tonic immobility. AB - Unilateral microinjection of carbachol (1 microgram) into the dorsal parabrachial region (PBR) of conscious guinea pigs produced a 100% increase in the duration of restraint-induced tonic immobility (TI) episodes. In another group of animals with a subcutaneous electrode introduced into the thigh region, microinjections of equivalent doses of the same drug in similar sites also significantly reduced the motor defense and vocalization responses elicited by the application of a noxious electrical stimulus to the skin. Both effects were blocked by pretreatment with atropine. The possible simultaneous activation of mechanisms modulating TI and the response to a noxious stimulus may be of adaptive importance since analgesia may reinforce immobility to permit the use of other defense mechanisms in a situation of prey/predator confrontation. PMID- 8240709 TI - Relationships between testing-induced alterations of hippocampal cholinergic activity and memory performance on two spatial tasks in mice. AB - Alterations in hippocampal cholinergic activity associated with different types and/or stages of learning were explored using measures of sodium-dependent high affinity choline uptake (SDHACU) in the hippocampus of C57BL/6 mice. Animals were divided into 'active' subjects submitted to memory testing before being killed and 'quite' controls. 'Active' subjects were trained in a radial-arm maze on either a discrimination task (mixed Working Memory (WM)-Reference Memory (RM) task) or a Delayed-Non-Matching-To-Place task (more selective WM-task). In the discrimination task mice were sacrificed after either the 1st, 2nd, 3rd or 9th daily session of training and at intervals of either 30 s, 15 min, 24 h or 9 days post-test. In the DNMTP-task all subjects of the 'active' groups were sacrificed after 12 days of training at either 30 s or 24 h post-test. Results showed that: (1) Both types of training induced an immediate (30 s post-test) increase of hippocampal SDHACU as compared to 'quite' control condition. (2) In the discrimination task, this immediate increase in SDHACU was followed by a decrease leading to a long-lasting (24 h and 9 days) inhibition of this cholinergic marker. This secondary decrease in SDHACU occurred earlier (15 min post-test) at the end (9th session) than at the beginning (1st-3rd sessions) of training. Thus, as training progressed there was a shortening of the testing-induced cholinergic activation. (3) By contrast, in the DNMTP-task, SDHACU was still increased at the interval of 24 h following the last session of DNMTP-training. (4) The amplitudes of both the immediate (30 s) increase and subsequent secondary (15 min) decrease in SDHACU after the last (9th) session of discrimination training were significantly related to the rate of acquisition and behavioural profile for individual animals. Subjects that had displayed better response accuracy across the 9 days of training exhibited the highest SDHACU at 30 s post-test and the lowest at 15 min post-test. These results are discussed in the context both of previous findings on the effects of training on cholinergic activity, and of contemporary models of hippocampal function. It is suggested that (1) an increase in hippocampal cholinergic transmission during testing would facilitate the acquisition of a 'relational' kind of informations (spatial WM and RM); (2) the post-training consolidation (spatial RM) of information would be facilitated by a decrease and long-lasting inhibition of hippocampal cholinergic activity. PMID- 8240710 TI - Differences in the recovery rate of a learned forelimb movement after ablation of the motor cortex in right and left hemisphere in white rats. AB - After ablation of the motor cortex contralateral to the preferred limb, rats were forced to use this limb in grasping food out of a horizontal tube by restraining the non-preferred foreleg with a bracelet. In a right-handed rat the motor behaviour is less impaired and the recovery is better after a left hemisphere lesion in comparison with a left-handed rat after a right hemisphere lesion. This difference is primarily the time needed to use the limb in reaching for seed (testperiod 3) and the total time needed for 10 successive seizings of seed (testperiod 5). Surprisingly, the differences in the course of recovery do not correlate with the degree of preoperative limb preference: the initially ambidextrous rate also show the same differences in results of the motor cortex ablation in left and right hemisphere. However, in the group of initially consistent ambidextrous rats, after 10 weeks of tests with a restrained forelimb, the testing under unrestrained free conditions shows a gradual decrease in the induced limb preference and a shift to the use of the foreleg contralateral to the intact hemisphere, while in contrast the initially consistent left- or right handed rats preserve the limb preference under the unrestrained testing conditions. Therefore the degree of initial preference still influences the choice of limb after motor cortex ablation and intensive training to use the 'damaged' limb. PMID- 8240711 TI - Structure of the rat's behaviour in the hot plate test. AB - The objectives of the study were: (i) to elaborate an ethogram of the rat's behaviour during the hot plate test, (ii) to analyse the rat's responses to the nociceptive heat stimulus by using ethological techniques, and (iii) to discern the suitable pattern or patterns to evaluate the rat's reaction to the noxious stimulus. Male Wistar rats (n = 30) were employed, and one hot plate test was carried out for each rat. The platform (Socrel DS37) was maintained at 55.0 +/- 0.5 degrees C, and the exposure time was 45 s. Behaviour was videotaped and analysed by an ethogram and a software package. Every pattern parameter (frequency, duration and latency for the first occurrence) was quantified. Cluster and Markovian analyses were also employed, whereby a dendrogram and a Markov chain were obtained, respectively. The ethogram was composed of twelve patterns: immobile-sniff, walk-sniff, rearing, forepaw-licking, hindpaw-licking, face-washing, body-cleaning, stamping, hindleg-withdrawal, leaning posture, jumping off, and freezing. Cluster analysis indicated that the most frequent patterns could be classified into three categories: sniffing responses (immobile sniff, walk-sniff), primary noxious-evoked patterns (forepaw-licking, hindpaw licking, stamping), and escape noxious-evoked responses (leaning posture, jumping off). Hindleg-withdrawal would be an independent noxious-evoked pattern, and freezing an alerting behaviour. The ethological information could be useful to better elucidate effects on rats' behaviour of drug treatment or other phenomena during the hot plate test. PMID- 8240712 TI - Influence of gender and age on performance of rats in the elevated plus maze apparatus. AB - We have compared the performance of male and female Wistar rats at different ages (45, 60, 90, 120 and 150 days) in the elevated plus-maze test, a reliable animal model of anxiety. Up to 60 days of age, rats of both sexes exhibited a high number of entries and of time spent on open arms (50% or above). At 120 days of age or more, rats of both sexes characteristically exhibited a reduction in the number of entries and of the time spent on open arms (below 50%). Within the range of 60 and 120 days there are statistically significant sex differences. At 90 days of age male rats showed a marked switch in their performance in the apparatus, reaching levels of the latter stage, whereas in females it happened around 120 days. These results suggest an ontogenetic difference in rats that accounts for at least two distinct performances for rats placed in an elevated plus-maze. Gender effects were found in a certain range, suggesting caution on interpreting data obtained in rats within 60 and 120 days old. Also, the results obtained highlight the importance of carefully controlling animal age in studies using the elevated plus-maze. PMID- 8240713 TI - Extralemniscal co-activation is not indispensable for behavioral detection of auditory stimuli. AB - Thresholds for triggering summed auditory evoked responses (ERs) were measured in non-auditory (= extralemniscal--EL) nuclei receiving direct auditory projections from the lateral lemniscus. Primary EL ERs with onset latency of 3-6 ms reflecting activation of direct EL projections of lemniscal auditory nuclei were registered in caudal pontine reticular nucleus (CPRN), in deep layers of superior colliculus (SC) and in ventromedial hypothalamus (VMH). Secondary EL ERs (waves of EL ERs with onset latency above 10 ms) reflecting diffuse auditory EL co activation of the brain, were registered besides the above mentioned nuclei also in the medial amygdala (MA). Threshold sound intensities for evoking primary EL ERs in CPRN, SC and VMH, for secondary EL ERs in all extralemniscal nuclei tested, and for conditioned avoidance behavior in a two-way shuttle box, were compared mutually. There were no significant mutual differences among thresholds for inducing secondary EL ERs in all EL nuclei tested. Thresholds for evoking secondary EL ERs were lower than those for evoking primary EL ERs in deep layers of the SC, equaled to thresholds for primary EL ERs in the VMH and were higher than thresholds for primary EL ERs in the CPRN. The results suggest that auditory EL projections into SC and/or VMH (but not into CPRN) might represent the primary triggering source for secondary EL ERs in various extralemniscal nuclei. Although conditioning lowered the threshold intensities for inducing secondary EL ERs, the threshold sound intensity for triggering conditioned behavior was lower than the threshold for secondary EL ERs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240714 TI - Light deprivation produces accelerated behavioral recovery of function from neglect produced by unilateral medial agranular prefrontal cortex lesions in rats. AB - Environmental manipulations involving 48 h of light deprivation have been found to produce sparing of function from neglect produced by unilateral destruction of the medial agranular prefrontal cortex (AGm) in rats. The present study is an extension of these findings and examined whether light deprivation would produce recovery in subjects with demonstrated severe neglect produced by unilateral AGm destruction. The orientation behavior of the subjects, 25 left and 16 right AGm operates, was tested at 4 h post injection and if severe neglect was demonstrated, the subjects were placed into one of three environmental conditions: (1) 48 h of total light deprivation, (2) 48 h of continuous light, or (3) 48 h of a 12:12 light/dark cycle. The left vs. right hemisphere comparisons indicated that the left hemisphere operates were significantly more responsive by 4 h postsurgery than the right hemisphere operates, and only five of the right hemisphere operates demonstrated severe neglect. Therefore, the effects of environment on behavioral recovery of function were only examined in left AGm operates with severe neglect. The results indicated that 48 h of light deprivation could produce dramatic behavioral recovery in left AGm operates with demonstrated severe neglect, and that these effects were not produced by disruption of the light/dark cycle. PMID- 8240715 TI - Haloperidol exaggerates proprioceptive-tactile support reflexes and diminishes vestibular dominance over them. AB - Rats made immobile and cataleptic by haloperidol, a dopamine receptor blocker, maintain their static stable equilibrium by employing a variety of allied postural support reflexes. Under some test conditions, competition between such reflexes occurs, and in haloperidol-treated rats, unlike undrugged controls, proprioceptive-tactile stimuli appear to be dominant over vestibular stimuli. We investigated this relationship in rats by testing their air-righting with and without simultaneous contact of the tail on a wooden platform. The rats were lightly held in a supine position by the shoulders and pelvis, with or without tail contact on a small wooden platform 47 cm above the ground. Undrugged rats showed the normal pattern of righting which involves axial rotation with cephalocaudal recruitment whether the tail is contacting the platform or not. Upon release, the haloperidol-treated rats (2.5 mg/kg) gripped the platform with their tail, which interfered with the air-righting reflex. This demonstrates that in haloperidol-treated rats, the dominance of tactile-proprioceptive postural support reflexes over those triggered vestibularly. PMID- 8240716 TI - Purification and quantitative analysis of nucleic acids by anion-exchange high performance liquid chromatography. AB - A novel and reliable high-performance liquid chromatography (HPLC) method is described for the purification and quantification of double-stranded DNA. The nucleic acids may be obtained by polymerase chain reaction (PCR) or as restriction fragments from enzymatic cleavage; the separated products are devoid of contaminating material like agarose, ethidium bromide or non-specific DNA sequences. Because of the non-destructive nature of this HPLC procedure, the purified DNA is optimally suited for cloning experiments. The DNA separation by HPLC has major advantages when combined with reverse transcription (RT)-PCR. This is exemplified by analysis of the TNF-alpha mRNA obtained from endotoxin-elicited rat liver macrophages. If the standard procedure of Northern blotting is compared with the combination of RT-PCR and quantification of the PCR products by HPLC, it is obvious that the dynamic changes of tumor necrosis factor (TNF)-alpha mRNA synthesis are at least as precisely reflected with the RT-PCR/HPLC combination. The latter method is presented as a reliable and powerful tool for quantitative studies on gene expression. PMID- 8240717 TI - Identification and characterization of microcystin-LY from Microcystis aeruginosa (strain 298). AB - Two toxins, a main component A and a minor component B, were isolated from the freshwater cyanobacterium Microcystis aeruginosa (strain 298) and characterized in their chemical structure by amino-acid analysis, configurational analysis, by FAB-MS and 1H-NMR spectroscopy. The acid hydrolysate yielded for toxin A as constituent amino acids D-Ala, L-Leu, D-Glu, erythro-D-beta-Me-Asp and L-Arg, and for toxin B the amino acids D-Ala, L-Leu, D-Glu, erythro-D-beta-Me-Asp and L-Tyr. 1D and 2D 1H-NMR spectroscopy of the toxins A and B in DMSO-d6 allowed to characterize them as cyclic heptapeptides containing both the unusual beta-amino acid Adda (3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid) and N-methyldehydroalanine (Mdha) as additional constituent residues. Toxin A was found to correspond to the known and structurally well characterized microcystin LR and toxin B to microsystin-LY. The presence of this variant has already been proposed, but its primary structure could be confirmed in this study. PMID- 8240718 TI - Taxonomical classification of the guinea pig based on its Cu/Zn superoxide dismutase sequence. AB - Cu-Zn superoxide dismutase was purified from guinea pig (Cavia porcellus) liver up to electrophoretic homogeneity and its amino acid sequence was elucidated by automated Edman degradation of proteolytic fragments and mass spectrometry. The protein was classified as a typical mammalian cytosolic Cu-Zn superoxide dismutase by molecular mass, specific activity, amino acid sequence and N terminal acetylation. A dendrogram constructed from previously known vertebrate cytosolic Cu-Zn superoxide dismutase sequences reflects the commonly accepted taxonomy and phylogenetic relationships of the species, whereas the guinea pig sequence is similarly remote form muriform rodents, lagomorphs, equiforms and primates. The data appear incompatible with the assumption that the Caviomorpha with the representative Cavia porcellus form a common phylogenetic clade with the muriform rodents but rather have to be considered a distinct order of mammals. The degree of similarity of the sequences further suggests that the mammalian clade diverged into rodents, primates, lagomorphs and caviomorphs at about the same time. PMID- 8240719 TI - Generation of matrix-degrading proteolytic system from fibronectin by cathepsins B, G, H and L. AB - By their endoproteinase activities, cathepsins B, G, H and L can generate matrix degrading proteolytic system from fibronectin. All four cathepsins studied cleaved fibronectin in fragments that were either proteolytically active or activated after incubation at pH 7.4 and in the presence of Ca2+. The highest enhancement of the matrix protein-degrading activity was observed after a gelatin affinity chromatography of each digest. These results suggest that the effect of cathepsins at physiological pH in vivo may be enhanced by the activation of a matrix-degrading proteolytic system from fibronectin. PMID- 8240720 TI - Rifampicin resistance of Mu lysogenic strains by rpoB mutations. AB - The resistance of Escherichia coli against the antibiotic rifampicin is caused by mutations in the rpoB gene which alter the structure of the beta subunit of the DNA-dependent RNA polymerase. By insertion-mutagenesis with bacteriophage Mu rifampicin-resistant mutants that were believed to have a sensitive RNA polymerase had been generated. For closer analysis of this putative rpoB independent mechanism of resistance we cloned and sequenced the insertion sites from two of the Mu lysogens. One of them showed > 95% sequence similarity with the phn locus of E. coli B, and was mapped between nt 3824 and 3825 within the phnB gene. This positions the phn locus between 4397 and 4410 kb of the Kohara map of E. coli K-12. Since both analysed insertion sites differ in respect to each other and to the findings of previous work we determined the rifampicin sensitivity of the RNA polymerase of the Mu lysogens with partially purified enzyme. For all mutants the IC50 as a measure for sensitivity was significantly higher than for the parent strain. Sequence analysis of part of the rpoB gene (nt 1519 to 1716) revealed single point mutations in the investigated Mu lysogens. The rpoB mutation is necessary and sufficient for the observed resistance, while the prophage alone does not evoke resistance and has no synergistic effect together with the rpoB mutation. Our work supports the assumption that rifampicin like other hydrophobic molecules enters the cell via simple diffusion through the outer membrane with LPS being the main barrier.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240721 TI - Lysosomal enzyme activities in liver and sera from guinea pigs fed oil related to the toxic oil syndrome. AB - beta-N-Acetylglucosaminidase and beta-galactosidase activities were determined in serum and liver from guinea pigs fed "toxic oil" (related to cases of TOS) under different experimental conditions. The results obtained were compared with those of guinea pigs fed non "toxic oil" (case-unrelated oil; controls 1) and animals fed no oil (controls 2). In serum, both activities were significantly increased after all treatments with case-related oil as compared with controls 1 and 2. In the liver, beta-galactosidase activity did not show significant differences in any of the treatments when compared with controls 2. However, NAG activity decreased significantly after 7 days of treatment with non-heated oil--either case-related or not--when compared with controls 2; it also decreased significantly after 28 days of treatment with heated case-unrelated oil, both with respect to controls 2 and the animals fed case-related oil. Liver weights tended to increase in the animals fed oil--toxic or not--with respect to those of the livers from untreated animals. Morphologically, a slight vacuolization of the hepatocytes was observed in some of the samples from the treated animals. PMID- 8240722 TI - Methotrexate in asthma. A safety perspective. AB - The inflammatory process underlying bronchial asthma is well established and has prompted clinical interest in nonsteroidal anti-inflammatory forms of treatment. Although unproven, it has been suggested that effective treatment of allergic inflammation may prevent long term consequences of asthma and avert deterioration in pulmonary function. Methotrexate has potent anti-inflammatory actions, even at low doses, and was judged to be a suitable candidate drug for asthma treatment if it could demonstrate an acceptable tolerability profile. Low dose methotrexate has been investigated in both noncomparative studies and in placebo-controlled studies of severe asthma. In general, such studies have suggested that methotrexate may have steroid-sparing benefits coupled to generally mild adverse events; although adverse effects were not of a serious nature they were observed in up to one-third of patients. Rare but potentially life-threatening adverse effects involving the pulmonary, hepatic and haematological systems remain of particular concern. Methotrexate should therefore be considered as an adjunct to high dose inhaled corticosteroids in patients who require more than 10mg of prednisolone daily, and who experience severe and unacceptable steroid-related adverse effects. Treatment should only be initiated by physicians with experience in the use of the drug, and the relevant safety parameters should be closely monitored. PMID- 8240724 TI - Cancer therapy and oral mucositis. An appraisal of drug prophylaxis. AB - Oral mucositis as a consequence of cytotoxic therapy is a major cause of morbidity in cancer patients. Cancer therapy-induced tissue damage leading to mucositis can occur through either direct or indirect stomatotoxicity. Once mucositis has occurred, treatment consists of measures to palliate symptoms. The prevention of cancer therapy-induced oral mucositis is less standardised. Numerous drugs have been used as prophylactic agents to prevent chemo- and radiotherapy-induced mucositis. Controlled trials have shown some degree of prophylactic efficacy for sucralfate, chlorhexidine and benzydamine. Positive but non-placebo-controlled trials requiring more study have been conducted with dinoprostone (prostaglandin E2), silver nitrate, beta-carotene, pentoxifylline and lozenges containing polymixin B, tobramycin and amphotericin B. Current studies have shown a lack of efficacy with allopurinol and granulocyte colony stimulating factor (G-CSF). Nonpharmacological methods such as oral cryotherapy and helium-neon laser treatments have shown some promise. At the present time no agent has been shown to be uniformly efficacious and can be accepted as standard therapy. Additional studies combining several agents or incorporating nonpharmacological manoeuvres for mucositis prevention are needed. PMID- 8240723 TI - Antiepileptic drugs. A review of clinically significant drug interactions. AB - Approximately 20 to 30% of patients with active intractable epilepsy are commonly treated with polytherapy antiepileptic drug regimens, and these patients may experience complicated drug interactions. Furthermore, because of the long term nature of treatment, the possibility of drug interactions with drugs used for the treatment of concomitant disease is high. Classically, clinically significant drug interactions, both pharmacokinetic and pharmacodynamic, have been considered to be detrimental to the patient, necessitating dosage adjustment. However, this need not always be the case. With the introduction of new drugs (e.g. vigabatrin and lamotrigine) with known mechanisms of action, the possibility exists that these can be used synergistically. The most commonly observed clinically significant pharmacokinetic interactions can be attributed to interactions at the metabolic and serum protein binding levels. The best known examples relate to induction (e.g. phenobarbital, phenytoin, carbamazepine and primidone) or inhibition [e.g. valproic acid (sodium valproate)] of hepatic monoxygenase enzymes. The extent and direction of interactions between the different antiepileptic drugs are varied and unpredictable. Interactions in which the metabolism of phenobarbital, phenytoin or carbamazepine is inhibited are particularly important since these are commonly associated with toxicity. Some inhibitory drugs include macrolide antibiotics, chloramphenicol, cimetidine, isoniazid and numerous sulphonamides. A reduction in efficacy of antibiotic, cardiovascular, corticosteroid, oral anticoagulant and oral contraceptive drugs occurs during combination therapy with enzyme-inducing antiepileptic drugs. Discontinuation of the enzyme inducer or inhibitor will influence the concentrations of the remaining drug(s) and may necessitate dosage readjustment. PMID- 8240725 TI - Clinically important ocular reactions to systemic drug therapy. AB - Many systemically administered drugs produce ocular adverse effects. Fortunately, relatively few are capable of causing significant, irreversible visual impairment. It is the responsibility of every clinician when prescribing systemic therapeutic agents to be aware of potential adverse ocular reactions, to appreciate their significance, and to inform the patient of the potential risks of treatment. In instances where serious adverse reactions relate to the cumulative effects of prolonged treatment, it is the responsibility of the prescribing physician to institute appropriate methods of visual screening. In this respect, it is most important to obtain the necessary individual baseline measurements before treatment is commenced. Chloroquine retinopathy is probably the most feared of all adverse ocular reactions to systemic drug therapy. However, it occurs only rarely if the daily dosage of chloroquine does not exceed 250mg. Regular screening using automated perimetry is mandatory if prolonged therapy is contemplated. Amiodarone almost inevitably produces corneal deposits. These rarely produce symptoms, and resolve upon withdrawal of the drug. Optic neuropathy has recently been described with amiodarone. Systemic anticoagulant therapy may be associated with intraocular hemorrhage in patients with pre existing disciform macular degeneration, and such agents should be used with caution in affected individuals. Systemic corticosteroids produce posterior subcapsular cataracts in susceptible individuals which may profoundly affect visual acuity. Although elevated intraocular pressure may also result from systemic therapy, the relationship between the pressure rise and development of glaucomatous changes remains unclear. Ethambutol may produce optic neuropathy if the daily dosage exceeds 15 mg/kg. The changes are usually reversible within a few weeks of stopping treatment. High doses of tamoxifen may produce a maculopathy with loss of visual acuity, if given for prolonged periods. The risk must be weighed against the benefits of treatment. Patients receiving more than 800 mg/day of thioridazine have developed retinopathy, which is usually reversible if detected early enough. Tricyclic antidepressants and other agents with anticholinergic properties may cause disturbances of accommodation and pupillary dilatation. The latter may rarely precipitate acute angle closure glaucoma in susceptible individuals. PMID- 8240726 TI - Clinical use of cocaine. A review of the risks and benefits. AB - Following the isolation of cocaine from the extract of coca leaves in the late 1700s by Albert Neiman, the local anaesthetic properties of the drug have been evaluated. The anaesthetic effect of cocaine is believed to be the result of reversible blockade of nervous impulse conduction by the prevention of sodium ion movement within the cell membrane. The many undesired effects, however, are the result of adrenergic stimulation by means of prevention of noradrenaline (norepinephrine) uptake. The clinical use of cocaine in the modern era is associated primarily with surgical procedures involving the manipulation of mucous membranes, particularly those of the upper respiratory tract. Recently, based on problems with the potential for illicit use and adverse effects, alternative regimens have been investigated. In some settings these have been superior to cocaine. Further studies comparing various anaesthetic regimens are warranted. PMID- 8240727 TI - Features, prevention and management of acute overdose due to antidiabetic drugs. AB - Hypoglycaemic medication forms a disparate group of therapeutic compounds including insulin, the sulphonylureas and biguanides. They are all designed to prevent hyperglycaemia and in general are well tolerated. Careful prescribing practice and patient education by the physician can do much to reduce the risk of adverse effects from diabetic therapy. However, the presentation of adverse effects, together with accidental and non-accidental overdose, is a frequent clinical problem. Furthermore, the possible impairment of hypoglycaemic awareness in patients prescribed human insulin has added complexity to diabetic management. The cardinal features of insulin overdose are hypoglycaemia and hypokalaemia. The sulphonylureas predominantly cause hypoglycaemia, while the biguanides may precipitate lactataemia and acidosis. Recognition of hypoglycaemia is therefore crucial in avoidance of toxicity. Intravenous dextrose is the mainstay of therapy following gut decontamination (for the oral agents). The efficacy of glucagon is dependent on hepatic glycogen stores and should therefore be used with caution. Diazoxide is not recommended. More recently, octreotide has been shown to be effective in sulphonylurea overdose. Patients should be admitted and monitored with serial blood sugar measurements for a minimum of 1 to 2 days as clinically warranted. PMID- 8240728 TI - Does proof of casualty ever exist in pharmacovigilance? AB - Identifying the adverse effects of drugs, thus transforming adverse events into adverse drug reactions, is a useful and necessary but complicated task. Objective proof of a causal relationship between a drug and a specific event is quite exceptional. In most cases, this relationship remains subjective and is no more than inner conviction. Several means are at our disposal to achieve causal assessment: spontaneous reporting, clinical trials, cohorts with and without controls, and case-control studies, with each having advantages and limitations. The search for casualty in pharmacovigilance is a necessary scientific goal, but a high degree of suspicion may be all that is necessary to withdraw a drug from the market if it is suspected of causing serious adverse effects. PMID- 8240729 TI - Vaccination in Third World countries. PMID- 8240730 TI - Tumor immunology: present perspectives. PMID- 8240731 TI - Tumor-rejection antigens recognized by T lymphocytes. AB - Generations of immunologists have been searching for evidence to confirm the tantalizing notion that tumor-rejection antigens exist. If found, the ultimate reward was the possibility that these molecules might be used to induce tumor specific immunity and effect tumor rejection. Until recently rewards have been few and far between. That is changing. The immediate rewards have become more satisfying and the ultimate reward almost palpable. PMID- 8240732 TI - Potential immunogenicity of oncogene and tumor suppressor gene products. AB - The immunogenicity of viral oncoproteins has been established beyond doubt. Cytotoxic T lymphocytes directed against viral oncogene products can eradicate large established tumor masses. This stage has not yet been reached for cellular oncogene and tumor suppressor gene products, but T cells have been raised against MHC-binding peptides encoded by both mutant and wild-type alleles of the ras oncogene and the p53 tumor suppressor gene. In addition, T cells specific for joining region peptides of abnormal fusion proteins resulting from chromosome translocation in tumor cells have been generated. Some of these peptides are processed in cells infected with, for example, vaccinia-ras, but direct anti tumor effects of peptide specific T lymphocytes remain to be demonstrated. PMID- 8240733 TI - Action of natural killer cells and macrophages in cancer. AB - According to the results obtained in several types of experiments the immunological anti-tumor effects that reflect the clinical situation involve T lymphocytes. The products of natural effector cells can contribute to anti-tumor immunity as part of the cytokine network. In addition, some cytokines can act on the tumor cells by influencing directly their proliferative capacity and/or through modification of their phenotype. In the tumor tissue the interaction between malignant cells and host cells on the one hand, and between the various types of host cells on the other hand, is complex. The participating mechanisms can act in both directions, with the result that the outcome of the balance may be either inhibition or stimulation of tumor growth. PMID- 8240734 TI - Non-antibody immunotherapy of cancer. AB - Immunotherapeutic agents have become important additions to the list of anti cancer treatments; further progress will require more information about how the immune system can respond to tumor cells. Tumor cells can avoid eliciting an immune response through several mechanisms. Understanding these mechanisms may suggest strategies to manipulate tumor cells so that they trigger an effective immune response. Another strategy being developed is to improve the therapeutic index of cytokines so that better anti-tumor effects can be elicited with less toxicity. PMID- 8240735 TI - Monoclonal antibodies in cancer therapy. AB - A review of the clinical trials of antibody based cancer therapies reveals that this approach can, in rare cases, induce complete remissions in individual patients with cancer. Since these trials have usually involved patients with large tumor masses, tumor cell inaccessibility is probably a major reason for the prevailing failures. Minimal residual disease, the stage when tumor cells are few and dispersed, should therefore be a more promising target for therapeutic antibodies. This hypothesis is supported by a prospective randomized trial on patients with resected Dukes C colorectal carcinoma that resulted in increased survival and prolonged recurrence-free intervals. Thus, in addition to strategies designed to produce more effective, human-derived reagents, efforts need to be concentrated on directing passive antibody therapy towards the appropriate target. PMID- 8240736 TI - Induction of B cell tumor dormancy by anti-idiotypic antibodies. AB - Long-term dormancy of murine B-cell lymphomas can be experimentally induced by immunizing the host with the idiotype expressed on the tumor. Interaction of the cells with anti-idiotype antibodies is sufficient to induce and maintain the dormant state. The growth of lymphoma cells interacting with anti-idiotype antibodies is arrested and they undergo dramatic changes in their morphology, cell-cycle status and oncogene expression. Regrowth of a tumor after long-term dormancy results from the emergence of a tumor cell variant that no longer responds to the antibodies with growth inhibition. These data demonstrate the feasibility of reversing a malignant phenotype of cells by specific growth arrest signals and suggest new approaches for therapeutic intervention in cancer. PMID- 8240737 TI - Transplantation. Antigen, co-stimulators and cytokines: regulators of allogeneic interactions. PMID- 8240738 TI - Monoclonal antibodies for the induction of transplantation tolerance. AB - Non-lytic antibodies to CD4 and certain other T cell adhesion receptors can guide the immune system to become tolerant to foreign antigens, and to regain tolerance in autoimmunity. Tolerance is maintained lifelong through the action of regulatory T cells that in turn can influence naive T cells to acquire the same regulatory properties. A fuller understanding of the molecular basis of infectious tolerance could lead to the design of better immunosuppressive protocols. PMID- 8240739 TI - The induction of tolerance to alloantigens using MHC class I molecules. AB - The identification of approaches that allow the adult immune system to be manipulated to facilitate the induction of tolerance to alloantigens is important for the development of more specific immunosuppressive therapy for use in transplantation. Novel strategies using genetic manipulation of recipient cells for the induction of tolerance to MHC class I antigens have been tested in experimental systems in the past year. These, together with new insights into the mechanisms responsible for maintaining peripheral tolerance, have advanced our progress in this challenging area of transplantation biology. PMID- 8240740 TI - Novel approaches to the control of graft versus host disease. AB - Graft versus host disease (GVHD) remains the major obstacle to the application of bone marrow transplantation across HLA barriers. Recent advances in our understanding of GVHD pathophysiology have resulted in the evaluation in animal models and in clinical trials of some novel approaches to avoiding and treating GVHD. Continued advances in our knowledge are likely to result in the clinical application of biological therapies to maximize graft versus leukemia effects and alloengraftment, while avoiding GVHD. PMID- 8240741 TI - Cytokines as mediators of organ graft rejection and tolerance. AB - The analysis of cytokines following organ transplantation continues to flourish as a major area of investigation for transplant biologists. Over the past year many papers have reported the use of both molecular and antibody-based tools to dissect the expression of cytokines during graft rejection in both experimental and clinical transplantation. Further, how the expression of cytokines is altered during the induction of tolerance has been investigated by several groups. PMID- 8240742 TI - Cytokine dysregulation as a mechanism of graft versus host disease. AB - Graft versus host disease (GVHD) remains the major complication of allogeneic bone marrow transplantation. T cells in the donor bone marrow recognize and react against host alloantigens and thereby initiate GVHD, but the precise mechanisms by which host tissues are damaged remain unclear. Recently, several convergent lines of evidence have suggested that inflammatory cytokines act as mediators of acute GVHD. Most of the clinical manifestations of GVHD may in fact be due to the dysregulated production of cytokines by T cells and other inflammatory cells. The complex interactions among cytokines and their cellular targets suggest that individual cytokines may play an important and distinctive role in the pathophysiology of GVHD. Perturbation of the cytokine network may function as a final common pathway of target organ damage, and the rapid onset of severe, acute GVHD can be considered a 'cytokine storm.' PMID- 8240743 TI - Is chronic graft versus host disease an autoimmune disease? AB - Chronic graft versus host disease continues to be a major problem following bone marrow transplantation even though the incidence and severity of acute graft versus host disease has been reduced. Recent investigations have suggested that the pathogenesis of chronic graft versus host disease is more similar clinically to an autoimmune disease than to acute graft versus host disease. PMID- 8240744 TI - Cancer. PMID- 8240745 TI - Transplantation. PMID- 8240746 TI - Early allograft function in canine single lung transplant. AB - An assessment of early graft function in canine single lung transplant recipients was made by analysing early postoperative radiographic progression, lung perfusion, bronchial patency and bronchial anastomotic wound healing and histopathology of the allografted lung. Eighteen mongrel dogs weighing 15kg on average were used. Donor lung bloc with a generous atrial cuff, the pulmonary artery and left bronchus were taken and flushed with Euro-Collins solution which implanted in the pneumonectomized recipient dog. Anastomosis was done with the atrium, pulmonary artery and bronchus in that order. To assess an early graft function, a protocol for a grading system was designed into the chest roentgenogram, lung perfusion scan, bronchial patency and histopathologic progression of the bronchial anastomosis and allografted lung (Table 1). The results were obtained as follows: Radiographically, clear to infiltrate was seen in 67% (8/12), 33% (5/15), 30% (3/10) and 33% (2/6) on postoperative day 0, 1, 2 and 3 respectively. Lobar to total opacification was 33% (4/12), 67% (10/15), 70% (7/10) and 67% (4/6) on days 0 to 3 (Table 2). Perfusion scan showed normal to mild defect in 43% (3/7) and moderate to severe defect in 57% (4/7) on day 0 and 100% (5/5) on day 2 (Table 3). The bronchial anastomotic site showed patent to mild stenosis in 100% (8/8) on day 0 and mild stenosis in 2/2 on day 9 bronchofiberscopically, and showed normal wound healing in 38% (3/8), cellular infiltration in 38% (3/8) and infarction in 25% (2/8) up to day 9 postoperatively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240747 TI - Distribution of HLA class I alleles and haplotypes in Korean. AB - The antigen (phenotype), gene (allele) and haplotype frequencies of HLA class I were analysed in 4,622 Koreans. With allele frequencies of over 0.05, the most frequent HLA-A,-B and -C antigens were A2, A24, A33, A11, A26, A31; B62, B51, B44, B54, B61, B35, B58, B60; Cw3, Cw1, Cw4, Cw7. Of these A2, A24, Cw1 and Cw3 were present in very high frequencies, respectively (0.3211, 0.2200, 0.2204, and 0.3737). The most common haplotypes with frequencies larger than 0.02 were A2 Blank, A33-B44, A33-B58, A11-B62, A24-B51, A24-B54, A2-B27, B54-Cw1, B58-Cw3, B51 Blank, B61-Cw3, B62-Cw4, B35-Cw3, B44-Blank, B60-Cw3, B27-Cw1, A2-Cw3, A2-Cw1, A24-Cw1, A33-Cw3, A26-Cw3, and A11-Cw4. A significant negative linkage disequilibrium was found for the haplotypes of A2-B7, A2-B44, A2-B58, A24-B13, A24-B27, A33-B54 and A33-B62, of which frequencies were larger than 0.003. The B C and A-C haplotypes which showed the significant negative linkage disequilibrium were B44-Cw1, B51-Cw1, B44-Cw3,B62-Blank, A2-Cw4, A2-Blank, A11-Cw3, A11-Blank and A33-Cw1 and had frequencies higher than 0.01. The findings presented here could be used per se to estimate the populational relationships or as the control data for HLA-disease investigation. Furthermore they could provide the scope for the definition of new antigens. PMID- 8240748 TI - Fragile site X chromosomes in mentally retarded boys. AB - The fragile X syndrome is a common X-linked mental retardation and autism, affecting females as well as males. The fragile site X chromosomes were studied in a series of 153 mentally retarded boys of unknown etiology to determine the frequency of fragile X syndrome, and to assess the feasibility of making a clinical diagnosis of the fragile X syndrome in young boys before cytogenetic results were known. The 10 boys (6.4%) were positive for fra (X) (q27). The phenotype of fra (X) (q27) positive patients were typical except one who also had sex chromosomal mosaicism. There were three pairs of siblings among the fra (X) (q27) positive patients. Frequency of expression of the fragile site was in 10 to 47 per cent of cells. In addition, 19 boys showed a previously unsuspected chromosomal abnormality. The frequency of the fragile X syndrome in the present study is not significantly different from those in Caucasians and Japanese population. The fragile X syndrome can be recognized by noting key aspects of family history as well as the clinical features in mentally retarded boys. PMID- 8240749 TI - Urodynamic evaluation in pediatric neurogenic bladder. AB - Urodynamic evaluation in pediatric neurogenic bladder is prerequisite for accurate prediction of prognosis and selection of appropriate treatment modality. We classified normal, hyperreflexic and areflexic bladders by cystometry. Hyperreflexic bladder was subdivided into two groups with or without detrusor sphincter dyssynergia and areflexic bladder into low pressure and high pressure. Among 36 patients with myelodysplasia on whom we performed a videourodynamic study vesicoureteral reflux was found in three of 7 patients with hyperreflexia with detrusor sphincter dyssynergia, two of 6 patients with hyperreflexia without detrusor sphincter dyssynergia and five of 10 patients with areflexia with high pressure. Low pressure areflexic bladder didn't show vesicoureteral reflux. When we evaluated 39 pediatric neurogenic bladder patients with a mean 25.5 follow-up months period, upper tract deterioration was improved and normal findings persisted in all the compliant patients to treatment. In the noncompliant group initial abnormal upper tract findings persisted or aggravated, especially in hyperreflexic bladder with dyssynergia and high pressure areflexic bladder. One of the three hyperreflexic bladders with dyssynergia and two of six high pressure areflexic bladders received augmented cystoplasty. These data show that urodynamic evaluation in pediatric neurogenic bladder provides excellent criteria for selecting high risk groups in pediatric neurogenic bladder. PMID- 8240750 TI - Corporeal blood gas changes according to duration of drug-induced prolonged erection. AB - The corporeal blood gas changes in accordance with the duration of the prolonged erection which developed after intracorporeal pharmacotherapy with papaverine and phentolamine were investigated in 62 impotence patients. The picture of the corporeal blood taken from 15 psychogenic impotence patients (a control group) at 10 minutes after intracavernous injection when they showed full erections was arterial but there was pCO2 rise and pH drop compared to femoral artery blood taken simultaneously. As the erection lasted longer, significant gas changes of the cavernous blood began to appear (p < 0.0001): increase in pCO2 and decrease in pO2 from 4 hours, decrease in pH from 5 hours, decrease in O2 saturation from 6 hours. Erections lasting for more than 16 hours showed significantly worse hypoxia (p < 0.05). Therefore, to prevent hypoxia and metabolic acidosis, drug induced prolonged erection would be better decompressed before it lasts for more than 4 hours. PMID- 8240751 TI - Metabolic alkalosis induced by plasmapheresis in a patient with systemic lupus erythematosus. AB - We report a patient with systemic lupus erythematosus (SLE), who had developed metabolic alkalosis during plasmapheresis. The metabolic alkalosis could be promptly corrected by reducing the amount of citrate load. The development of metabolic alkalosis can be explained by the citrate load during plasmapheresis. Careful monitoring of acid base status is mandatory in patients with limited renal function and the reduction of citrate load may be advisable in plasmapheresis. PMID- 8240752 TI - Primary breast lymphoma. AB - We report the case of a 20-year-old female with lymphoma of the breast. Mammography showed an asymmetric pattern of confluent densities without any discrete mass. Sonography revealed diffuse heterogenous echoic mass intermingled with low-and medium level echoes. We present the clinical, radiographic and histologic features of primary breast lymphoma with a brief review of the literatures. PMID- 8240753 TI - Leigh's disease involving multiple organs. AB - Leigh's disease is a rare progressive neurological disorder that is characterized light microscopically by focal spongy necrosis in the brain and electron microscopically by mitochondriopathy. We report an autopsy case of Leigh's disease that showed abnormalities in the liver, kidney and skeletal muscle as well as the central nervous system. The patient was an 18-month-old girl who has carried a diagnosis of cerebral palsy ever since her birth to a 20-year-old mother. The baby was generally hypertonic and mentally retarded. She died of severe metabolic acidosis. Postmortem examination showed growth retardation, fatty liver, fatty kidney and soft brain. Brain section showed multifocal softenings in the brainstem, basal ganglia and periventricular areas. Microscopically increased capillaries with endothelial proliferation, vacuolar degeneration and mild gliosis were seen in the brain. The axons were relatively preserved. Liver and kidneys showed microvesicular fatty change. Myofiber degeneration of the skeletal muscle was also noted. Electron microscopic examination showed markedly increased mitochondria in the parenchymal cells of the brain, liver and kidney. The mitochondria showed round to ovoid ballooned appearance including electron-dense core-like structures and pseudoinclusions of glycogen granules. PMID- 8240754 TI - A case of juvenile form Pompe's disease manifested as chronic alveolar hypoventilation. AB - We describe a case of the juvenile form of Pompe's disease that presented as primary alveolar hypoventilation due to respiratory muscle involvement. This 17 year-old girl had been asymptomatic until this admission, although she had a delayed puberty. Arterial blood gas analysis, pulmonary function test as well as physical findings were compatible with chronic alveolar hypoventilation syndrome. Since she had lower extremity muscle weakness and pseudomyotonic discharge on electromyography a muscle biopsy was done, which revealed glycogen storage disease. The patient was managed successfully with nasal intermittent positive pressure ventilation. PMID- 8240755 TI - Peripheral neuropathy as a hypereosinophilic syndrome and anti-GM1 antibodies. AB - The acute peripheral neuropathy as one of hypereosinophilic syndrome is known to be a rare disorder. The authors experienced a dramatic case with acute peripheral neuropathy, hypereosinophilia in peripheral blood, and the positive anti-GM1 antibodies. The serum protein electrophoresis showed a diffusely increased gamma globulin region and the polyclonal gammopathy was found by the immunoelectropheresis. There was no evidence of inflammatory myopathy in vastus lateralis muscle. The sural nerve biopsy was compatible with vascular neuropathy, as there were a few myelin digestion chambers, mild perineuronal fibrosis, and perivascular lymphoplasmocytic infiltration with focal organizing thrombosis. The clinical response to prednisone therapy was excellent. PMID- 8240756 TI - Arterial thrombosis associated with nephrotic syndrome--a case report and review (adult cases in the English literature). AB - The thromboembolic complications of nephrotic syndrome are reasonably common, including spontaneous peripheral venous and/or arterial, pulmonary arterial, and renal venous occlusions. However, in comparison to the relatively high incidence of the venous thromboembolic complications with hypercoagulable status, arterial thromboses have been reported much less and it was only 20 cases in the English literature so far. Furthermore, the most cases were pediatric patients rather than adults. Therefore, this report describes an adult nephrotic cases complicated by superior mesenteric artery thrombosis leading to death via catastrophic hospital course. Also, we reviewed the literature in English regarding cases of arterial thromboses in adult nephrotic patients with special interest to locations of thrombosis, underlying histopathologic types of glomerulopathy, and use of steroids or diuretics before its development. PMID- 8240757 TI - Central core disease--a case report. AB - Central core disease is a rare congenital myopathy characterized by the formation of "cores" that consist of abnormal arrangement of myofibrils inside the myofibers. We report a 5-year-old Korean girl who showed a fairly typical clinical course of non-progressive muscle weakness. Electrodiagnostic studies showed low-amplitude polyphasic electromyograph and normal nerve conduction velocity. Gastrocnemius muscle biopsy showed central cores in over 80% of the fibers on H&E section. Histochemistry revealed deficient or absent mitochondrial enzyme in the cores and type I predominance. Ultrastructurally both structured and non-structured cores were found separately or simultaneously in one fiber. This case is the first report in the Korean literature. PMID- 8240758 TI - Nitrogen dioxide and respiratory illness in children. Part I: Health outcomes. AB - We have carried out a prospective cohort study to test the hypothesis that exposure to nitrogen dioxide increases the incidence and severity of respiratory infections during the first 18 months of life. Between January 1988 and June 1990, 1,315 infants were enrolled into the study at birth and followed with prospective surveillance for the occurrence of respiratory infections and monitoring of nitrogen dioxide concentrations in their homes. The subjects were healthy infants from homes without smokers; they were selected with stratification by type of cooking stove at a ratio of four to one for gas and electric stoves. Illness experience was monitored by a daily diary of symptoms completed by the mother and a telephone interview conducted every two weeks. Illnesses with wheezing or wet cough were classified as involving the lower respiratory tract; all other respiratory illnesses were designated as involving the upper respiratory tract. Exposure to nitrogen dioxide was estimated by two week average concentrations measured in the subjects' bedrooms with passive samplers. This analysis is limited to the 1,205 subjects completing at least one month of observation; of these, 823 completed the full protocol, contributing 82.8% of the total number of days during which the subjects were under observation. Incidence rates for all respiratory illnesses, all upper respiratory illness, all lower respiratory illnesses, and lower respiratory illness further divided into those with any wheezing, or wet cough without wheezing, were examined within strata of nitrogen dioxide exposure at the time of the illness, nitrogen dioxide exposure during the prior month, and type of cooking stove. Consistent trends of increasing illness incidence rates with increasing exposure to nitrogen dioxide were not evident for either the lagged or unlagged exposure variables. The effect of nitrogen dioxide exposure on illness occurrence during at-risk intervals of two weeks' duration was examined using the generalized estimating equation approach. In these multivariate analyses, none of the odds ratios was significantly elevated for unlagged nitrogen dioxide exposures, lagged nitrogen dioxide exposures, or stove type. Duration of illness was assessed in relation to the same exposure variables; illness duration and nitrogen dioxide exposure were not associated. We have found that indoor exposure to nitrogen dioxide is associated with neither the incidence nor the duration of respiratory illnesses. The study was designed to have sufficient power to detect effects of nitrogen dioxide exposure of magnitudes previously reported and in a range relevant to public health concern; the lack of association cannot be attributed to potential bias from misclassification of outcome or exposure.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8240759 TI - Nitrogen dioxide and respiratory illness in children. Part II: Assessment of exposure to nitrogen dioxide. AB - Repeated measurements of nitrogen dioxide were obtained from 1988 to 1991 in the homes of 1,205 infants living in Albuquerque, NM. Passive diffusion samplers were used to obtain a series of two-week integrated measurements from the home of each infant for use in a cohort study of the relation of residential exposure to nitrogen dioxide and respiratory illnesses. Information on stove use and time spent inside the residence was collected at two-week and two-month intervals, respectively. During the winter, in the bedrooms of homes with gas cooking stoves, mean nitrogen dioxide concentrations were 21 parts per billion (ppb); mean concentrations in the living room and kitchen were 29 ppb and 34 ppb, respectively. In homes with electric cooking stoves, the mean bedroom concentration was 7 ppb during the winter. Lower indoor concentrations were observed during the summer in homes with both gas and electric stoves. On average, infants spent approximately 12.3 hours per day in their bedrooms, 7.3 hours in the living rooms, 35 minutes in the kitchens, and 3.8 hours out of their homes. (As a condition of participation, none of the infants spent more than 20 hours per week in day care outside of their homes). The mean time infants spent in the kitchen during cooking was approximately nine minutes per day. We tested whether exposures of infants living in homes with gas stoves could be reasonably estimated by measurements in the bedroom in comparison with time-weighted average concentrations based on time-activity data and simultaneous nitrogen dioxide measurements in the kitchen, living room, and bedroom. In 1,937 two-week intervals from 587 infants, 90% of time-weighted exposure (on the three-level classification used in this study) estimates were in agreement with estimates based on bedroom concentrations alone. The agreement of the time-weighted nitrogen dioxide exposure estimates with the bedroom concentrations is attributed to limited amounts of cooking stove use (the mean was 29 minutes per day), small room-to-room differences in nitrogen dioxide concentrations (the mean kitchen bedroom difference was 12 ppb), and the relatively large proportion of time that infants spent in their bedrooms. PMID- 8240760 TI - Personality profiles of nurses: a comparison between Australian and US research findings. AB - The findings from an analysis of the personality profiles of 349 registered nurses in New South Wales are reported. Comparisons were made between males and females and between nurses practicing in seven nursing areas. The NSW nurses' profile was compared with those of a group of registered nurses in the United States of America and with a United States adult female normative group. An exploration of the effects of time on personality was made by analysing data from the New South Wales registered nurse group and a group of student nurses in the same state. Differences were found in the personality profiles of all groups examined in the study. These findings and their nursing implications are discussed with reference to the effects of primary and secondary socialisation. PMID- 8240761 TI - Nurses in OR are more assertive than radiographers. AB - This study investigated two hypotheses: that there is a positive correlation between assertiveness and self-esteem, and that nurses rate lower than non-nurses on the constructs. A self-report survey incorporating scales for general assertiveness, situationally-specific assertiveness, global self-esteem and differentiated self-esteem was utilised. For the sample of 83 operating room nurses and 81 radiographers, correlations significant at the p < .001 level between the respective scales supported the first hypothesis. Analysis of variance yielded no significant differences between the two groups on either self esteem measures or general assertiveness, with the nurses scoring significantly higher than radiographers on the situationally-specific assertiveness scale. Findings challenge the stereotype of the 'shrinking violet' nurse. Additional analysis revealed that the power component of differentiated self-esteem had a higher correlation with global self-esteem and with both assertiveness measures than any other component. Further, the operating room nurses were significantly more power-oriented than radiographers. The pre-eminence of power in the findings suggests that assertiveness and self-esteem are issues relating to the empowerment of nurses. PMID- 8240762 TI - The effects of All-RN and RN-EN staffing on the quality and cost of patient care. AB - This paper presents empirical evidence from a study that compared the quality and costs of patient care provided by two staffing regimes in an acute medical and an acute surgical ward in one hospital. As assessed by patient outcomes, there were few significant differences in either ward in the quality of care provided by all registered nursing staff and staff that was comprised of 80% registered nurses and 20% enrolled nurses. In the acute medical ward the all-registered nursing staff was more expensive per day and per hour for patient care adjusted for patient dependency; in the acute surgical ward the registered-enrolled nurse staff mix was more costly. The study found that nurses in both wards were confronted by continuously high workloads. Registered nurses reported that working with enrolled nurses (who were inexperienced in acute medical and surgical nursing) exacerbated their workload and, to a lesser extent, made their work more stressful. Large scale, long term studies are necessary to clarify the issues of cost effectiveness of RN-EN mix and workloads. PMID- 8240763 TI - IndoChinese women's breastfeeding practices following immigration to Sydney: a pilot study. AB - Traditionally, most IndoChinese women breastfeed their children for a prolonged period. Recent studies indicate that there is a reduction in the rate and duration of breastfeeding by IndoChinese women following immigration to Western countries. The purpose of this pilot study was to compare pre- and post-migration attitudes of 20 IndoChinese women towards infant feeding. Triangulated methodology was used for data collection and frequency distribution and measures of association were used to analyse the quantitative data. Content analysis identified patterns and themes in the qualitative results. Findings revealed that following migration to Australia the women experienced significant sociocultural problems. Recommendations include in-depth studies of ethnic groups and 'tailor made' education to promote breastfeeding. PMID- 8240764 TI - Structural and perceived medical dominance: a study of barriers to nurses' workplace satisfaction. AB - Current literature on the sociology of healthy views medical dominance as a structural feature of the health division of labour and a body of literature has developed exploring the structural components that contribute to the subordination of the allied health professions. In this paper, Friedson's four factor definition of medical dominance was employed to provide a framework for assessing the degree to which structural medical dominance of the nursing profession impinges on nurses' perceptions of dominance and how structural and perceived medical dominance affects nurses' workplace satisfaction. A 69 item questionnaire covering aspects of doctor/nurse, doctor/patient, nurse/patient and nurse/hospital administration relationships was developed. One hundred and thirty nurses completed the questionnaire in which they ranked their own level of professional satisfaction and their perceptions of level of doctors' professional satisfaction. Results indicated that nurses were dissatisfied with many aspects of their work environment, i.e. pay and working conditions and experienced very high levels of dissatisfaction with their professional status. Nurses perceived doctors as being highly satisfied and as having more control over a range of procedures intrinsic to the delivery of health care, and the medical profession to be a more powerful and higher status profession that was held in greater regard by the public and other allied health professionals than nursing. The implications of the perceived discrepancy in nurses' and doctors, workplace satisfaction are discussed in terms of the structural barriers created by medical dominance. Implications of the findings for nurse education and nurse practitioners are also discussed, together with suggestions for further research. PMID- 8240765 TI - How many hospitals have a nursing research policy? A Victorian survey. AB - Directors of nursing in 29 Victorian hospitals responded to a questionnaire which sought information on their hospitals' nursing research policies, structures and education and on the level of organisational support for nursing research. One quarter of the hospitals had nursing research policies, 45% expected nurses to be involved in research, 31% had nursing positions with primary research functions, 31% had nursing research committees and 50% provided some research education. Hospitals which had a written policy reported significantly higher levels of administrative support for nursing research and were more likely to have a designated nursing research position. It was concluded that the adoption of a policy on nursing research is associated with organisational support for nursing research in Victorian hospitals. PMID- 8240766 TI - Psychosocial influences in postnatal depression. AB - Changes in moods, behaviours and physical states following the birth of a baby are well known and generally accepted as common, expected and short-lived. It is also recognised that a number of women are severely affected postnatally by a psychotic illness that may require lengthy hospitalisation and treatment. Between these two extremes is the profoundly disturbing experience of postnatal depression of non-psychotic origin that may persist for many months, seriously disrupting the mother-baby and marital and family relationships. The extent and characteristics of PND are receiving attention from health care professionals who have been largely unaware of the problem and are poorly prepared to respond. Although many early reports support biological theories of this problematic disorder, recent studies have begun to examine psychosocial sources of strain. This paper, through drawing together evidence from stress research and maternal and infant development, presents a multifactorial model to describe postnatal depression. PMID- 8240767 TI - The effects of an education video on patients' requests for postoperative pain relief. AB - A patient education videotape called if it hurts, tell us was used to provide one of three groups of 20 preoperative patients with information about pain control and to encourage them to be assertive in their requests for postoperative pain relief. There were no significant differences in the levels of pain or requests for analgesia between the group which had been specifically encouraged by the education video to seek pain relief and the groups which had not seen the video. Patients in all three groups generally experienced a good deal of pain following surgery, but they were not dissatisfied with this experience and did not take available action to eliminate all their pain. While the literature suggests that improvements in postoperative pain management might be achieved by better staff education, patients' reluctance to eliminate their pain should also be considered. PMID- 8240768 TI - The use of touch in nursing practice. AB - This paper examines the types of touch used in nursing practice and the effects of touch on the body and mind. The effects of two types of touch, therapeutic touch and massage, are evaluated by reviewing research studies conducted by nurses. PMID- 8240769 TI - Effect of environmental stress and surface treatment on resin-to-metal bonding. AB - This study compares the bond strength and durability of three metal surface treatments subjected to two types of environmental stress for both short- and long-term exposures. The luting resins Panavia and Comspan were applied to alumina-blasted, non-beryllium, nickel-chromium alloy coupons. Metal surface treatments consisted of either microscopic roughening by electrochemical etching, or one of two types of adhesives: a silanated silica coating (Silicoating) or a phosphate ester monomer (a component in the Panavia liquid). Shear bond strength was determined following short- or long-term exposure to either thermocycling in 6-60 degrees C water (2,672 cycles/7 days or 10,584 cycles/42 days) or storage in 37 degrees C water (7 or 42 days). Three-way ANOVA showed that both the type of environmental stress and the exposure time affected the bond strength of electroetched surfaces, but that only exposure time affected the two chemical adhesives (P < 0.05), regardless of the environmental stress used. In the short term, the silica/silane coated surfaces produced and maintained the higher shear bond strengths (15.9 +/- 2.3 MPa). However, after 42 days the silica/silane bonds decreased 30% (to 11.3 +/- 2.2 MPa), while the phosphate ester bonds were essentially unchanged (11.4 +/- 3.0 at 4 days, 10.4 +/- 2.2 MPa at 42 days). Electroetched bonds were the weakest and decreased by 18% between 7 and 42 days in water (8.8 +/- 1.2 to 7.2 +/- 3.0 MPa) and 27% after 42 days of thermocycling (7.2 +/- 2.8 to 5.3 +/- 1.8 MPa). PMID- 8240770 TI - Effects of oxalate on dentin bonding. AB - The purpose of this study was to determine how much topical treatment of dentin with potassium oxalate would interfere with subsequently placed dentin bonding resins. The results indicated that oxalate lowered the bond strengths of Superbond, All Bond and Scotchbond 2, but that normal bond strengths could be obtained for Superbond and All Bond after pumicing the oxalate-treated dentin. PMID- 8240771 TI - Quantitative microleakage of six dentin bonding systems. AB - This in vitro study evaluated the quantitative microleakage (ML) of (A) Gluma/Pekafill, (B) Tripton/Opalux, (C) Syntac/Heliomolar, (D) Prisma Universal Bond 2/Prisma AP.H., (E) Prisma Universal Bond 3/Prisma AP.H., and (F) Clearfil PhotoBond/Photo Clearfil Bright to cementum (dentin) by means of a dye-recovery spectrophotometric technique. Standardized cylindrical Class V preparations, 3 mm in diameter and 1.5 mm deep, were made on the facial surfaces of the roots of ninety extracted human maxillary and mandibular permanent canines and the teeth were restored with the recommended resin composite. The root apices were removed, Class V preparations made at the cut apices and restored with two coats of Copalite varnish and amalgam. The teeth were painted with two coats of nail varnish except for 1 mm surrounding the restorations prior to thermocycling x500 in 2% methylene blue solution. Root sections including the restorations were placed in 50% HNO3, the dye extracted and the dye concentrations determined spectrophotometrically. Standard solutions with known amounts of dye were prepared in 50% HNO3 and unrestored root sections placed in these. ML was expressed as micrograms dye/restoration and data analyzed by ANOVA and Duncan's test. The following results were obtained: (A) 3.67 +/- 2.18; (B) 2.76 +/- 1.71; (C) 2.57 +/- 0.97; (D) 2.56 +/- 2.51; (E) 1.21 +/- 0.24; (F) 0.49 +/- 0.20. Clearfil PhotoBond and Prisma Universal Bond 3 showed significantly less microleakage than other systems evaluated. PMID- 8240772 TI - Effects of dentin permeability and hydration on the bond strengths of dentin bonding systems. AB - This in vitro study evaluated the shear bond strengths of five adhesive systems bonded to dentin with and without simulated physiologic hydrostatic pressure. To simulate pulpal pressure, bonding procedures were done on extracted teeth connected to a hydration apparatus. Bond strengths were significantly less for each adhesive system under simulated physiologic conditions. Amalgambond had significantly higher bond strengths than Clearfil Photo Bond, Gluma, Scotchbond 2, or Tenure, either with or without simulated pulpal pressure. PMID- 8240773 TI - Shear strength and microleakage of light-cured glass ionomers. AB - Bond to dentin and marginal integrity of three light activated glass ionomer cements were evaluated, and compared to a traditional chemical-cured glass ionomer cement. Results indicated no significant difference in shear bond strength among the glass ionomer cements tested. However, there was a significant difference among glass ionomer cements in bond strength variability, with coefficients of variation ranging from 55 to 102%. A discrepancy between manufacturers' instructions and optimum curing times is one possible explanation for these results. There were also no significant differences in microleakage among glass ionomer cements. Most samples exhibited minimal or no microleakage. PMID- 8240774 TI - Natural and induced radiation caries: A SEM study. AB - Radiation caries is a well-known indirect side effect of head and neck radiotherapy. The initiation and progression of radiation caries were studied using an in situ model. The morphology of induced carious lesions was compared with that of natural radiation caries. Both natural and induced radiation caries showed the same patterns of decay; widespread areas with porosity of enamel, crater formation with exposure of subsurface enamel, preferential dissolution of prisms with hollowing out of prism cores, loss of large parts of surface enamel and loss of full enamel coverage exposing the underlying dentin. In irradiated patients most enamel slabs were severely demineralized within 6 weeks whereas in control subject the slabs showed no significant demineralization after 12 weeks. The model may offer an excellent opportunity to study preventive fluoride regimens because of the rapid induction of radiation caries and its similarity with natural lesions. PMID- 8240775 TI - A comparative study of three glass ionomer base materials. AB - This study compared the compressive and diametral tensile strength, compressive modulus, shear bond strength to dentin and resin composite, marginal gap between the base and tooth in Class V cavity preparation, and fluoride release of three glass ionomer base materials. In general, visible light-cured (VLC) base materials outperform the autopolymerizing glass ionomer base Ketac-Bond. The compressive modulus is significantly less for the VLC glass ionomer materials Vitrebond and Fuji LC compared to Ketac-Bond. The Fuji LC had greater diametral tensile strength, compressive strength, bond to dentin and bond to resin composite than Vitrebond or Ketac-Bond. The fluoride release of Vitrebond was greater than Ketac-Bond or Fuji LC. The marginal gap at the tooth/base interface was significantly less with the Fuji LC compared to the other two materials. PMID- 8240776 TI - Microleakage resistance of glutaraldehyde-containing adhesives. AB - This study evaluated the microleakage resistance of various glutaraldehyde containing dental adhesives in Class V preparations. The materials tested were Gluma Bond System (3 Step), Prisma Universal Bond 3, and Syntac. The Class V preparations extended just beyond the cemento-enamel junction of extracted human molars. The restored teeth were stored in tap water for 24 hours and then thermally stressed for 100 cycles. Marginal leakage was assessed by measuring the dye linear penetration. The results indicated that all three dentin bonding agents provided leak-proof seals under certain conditions. The sealing ability of the Gluma Bond System (3 Step) appeared to be significantly affected by the resin composite used in the restoration. PMID- 8240777 TI - Effect of aging on dentin bonding. AB - The purpose of this paper was to evaluate the effect of aging of dentin on the tensile bond strengths of four dentin bonding systems using proximal dentin of premolars extracted from young (aged from 9-21 years) and old (aged from 42-64 years) patients. Clearfil Linerbond System and Super Bond D-Liner showed higher bond strengths to both young and old teeth than those of Clearfil Photo Bond and Resto Bond 3. However, similar bond strengths to both young and old teeth for all four products were obtained despite the effects of aging. Furthermore, a SEM study of the site of bond failure revealed the persistence of the resin impregnated layer on the dentin surface, in spite of the difference of bonding system and dentin, indicating that adhesive failure tended to occur at the interface between the bonding resin and the resin impregnated layer. PMID- 8240778 TI - Shear bond strength and microleakage of All-Bond. AB - The shear bond strengths (SBS) were determined to enamel etched with Uni-Etch (Procedure A) or treated with the mixed Primer A and B (Procedure B), to dentin treated with the Dentin Conditioner (Procedure C) or etched with All-Etch (Procedure D), to amalgam treated with the mixed Dual Opaquer Base and Catalyst which was allowed to self cure (Procedure E) or light cured (Procedure F), of amalgam to dentin (Procedure G), to cast metal alloy (Procedure H), and to porcelain (Procedure I). The test specimens were stored in physiological saline at 37 degrees C for 24 hours, and the SBS determined by the application of a shear load directed at the base of the bonded composite cylinders at a crosshead speed of 0.5 mm/minute. The SBS were calculated and expressed in MPa. Standardized Class V preparations were made at the facial aspects of the roots of the teeth, treated with the Dentin Conditioner (Procedure J) or etched with All Etch (Procedure K). Quantitative microleakage (QM) was determined by means of a spectrophotometric dye-recovery method and expressed in microgram dye/restoration. The mean +/- SD of the SBS in MPa were: A: 18.3 +/- 2.1; B: 22.0 +/- 2.9; C: 17.2 +/- 3.6; D: 15.6 +/- 3.7; E: 12.9 +/- 1.9; F: 13.6 +/- 2.0; G: 13.0 +/- 3.5; H: 21.8 +/- 2.6; I: 8.2 +/- 2.8. The mean +/- SD of the QM in microgram dye/restoration were: J: 2.61 +/- 1.22 and K: 1.95 +/- 0.59. The analysis of the SBS data showed that the application of the Primer to enamel resulted in a significant increase in SBS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240779 TI - Bond strength of composite to etched and sandblasted porcelain. AB - In vitro tensile bond strengths of composite to porcelain treated by acid etching or sandblasting were measured. A 9.5% HF gel was applied for 30, 60, 150, or 300 seconds at 23 degrees C. Al2O3 particles were applied as wet-10.2, dry-33.5, dry 48.0, or dry-78.0 microns. Average roughness (Ra) was measured by profilometry before and after each treatment. Both longer etching time and sandblasting using Al2O3 with larger particle sizes produced increased surface roughness of porcelain. Etching with 9.5% hydrofluoric acid gel produced higher bond strengths than sandblasting with a series of Al2O3 particles. Etching longer than 60 seconds produced increased cohesive failures in porcelain, whereas most failures observed with sandblasted surfaces were adhesive. PMID- 8240780 TI - Fracture resistance of endodontically-treated cuspids. AB - The purpose of this in vitro study was to determine the optimum method for treating an endodontically treated cuspid prior to placement of a crown. Sixty extracted human maxillary and mandibular cuspid teeth were divided into groups of 10. In groups A and B, the crowns were removed at the CEJ and post amalgam cores were placed with #5 Paraposts (Group A) and #120 tapered Ticonium Endowels (Group B). The remaining groups received the following treatments: Group C--gutta percha endodontic fillings, Group D--bonded composite (Fluoro-Core) in the access preparations, Group E--#100 Ticonium Endowels, and Group F--untreated control group. All teeth were prepared for and received cemented gold crowns, were mounted in methyl methacrylate and were stressed at 45 degrees to failure in an Instron machine. Mean fracture strengths (Newtons) were: Group A--500, Group B- 530, Group C--520, Group D--510, Group E--680, Group F--580. Results indicated that when the coronal tooth structure was removed, there was no statistical difference (P < 0.05) between the Parapost and Endowel core buildup groups. Although Group E (intact crowns+Endowel posts) displayed higher mean fracture resistance than all other groups, there was no statistical difference (P < 0.05) due to the large range. PMID- 8240781 TI - Shear bond strengths of new enamel etchants. AB - Many dental adhesive systems now rely on simultaneous etching of enamel and dentin. Instead of the traditional 35-40% phosphoric acid, new etchants (including 10% maleic acid, 10% phosphoric acid, and oxalic acid/aluminum nitrate) are used. This in vitro study determined the shear bond strengths of resin composite to enamel which was etched with these agents according to manufacturers' instructions. The control etchant, 35% phosphoric acid, gave a mean bond strength of 24.5 MPa. 10% maleic acid and 10% phosphoric acid had significantly lower shear bond strengths (13.2 MPa). Gluma 1 & 2 Conditioner (oxalic acid/aluminum nitrate) had a mean bond strength of 6.3 MPa, which was significantly less than the other etchants. Bond strengths for each etchant were not affected by the type of unfilled resin used. PMID- 8240782 TI - Dentures for a 3-yr-old child with ectodermal dysplasia: case report. AB - A case of a 3-year-old boy with X-linked hypohidrotic ectodermal dysplasia is presented. Complete maxillary and mandibular dentures were made at 3 years and 2 months of age. The process for making the dentures is described. Obtaining enough patient cooperation and proper application of behavioral management techniques are critical factors for the procedure described in the article. PMID- 8240783 TI - The involvement of antigen processing in determinant selection by class II MHC and its relationship to immunodominance. AB - The T cell response in vivo to many whole proteins is focused on a limited number of possible determinants which can be termed immunodominant. Antigen processing for class II antigen presentation appears to play a major role in this selective process. With experimental evidence accumulated in our laboratory as well as others, we review several possible mechanisms involved in antigen processing responsible for selective or differential determinant expression. In particular, we discuss the determinant capture model in which MHC class II molecules initially capture large antigen fragments, such that bound determinants are protected from proteolysis by the MHC molecules and eventually become dominant while the flanking determinants are trimmed away. Such flanking determinants therefore become subdominant or cryptic. This mechanism underlies the capturing role of MHC molecules in the physiological processing of antigens. PMID- 8240784 TI - Sertoli-Leydig cell tumour of the ovary--a rare cause of virilization after menopause. AB - A case of Sertoli-Leydig cell tumour of the ovary causing virilization in a postmenopausal woman is presented. The patient had increasing facial hair growth, deepening of the voice, a dull pain in the lower part of the abdomen and enlargement of the clitoris. Laboratory investigations showed an elevated level of plasma testosterone. Considering an ovarian tumour the most likely cause of the above-mentioned findings, hysterectomy with bilateral salpingo-oophorectomy was performed. Both ovaries were macroscopically normal at operation, but pathological examination revealed a small well-differentiated Sertoli-Leydig cell tumour in the left ovary. Follow-up 3 months later showed decreasing signs of virilization and normalization of the hormone levels. PMID- 8240785 TI - Altered course of Plasmodium berghei infection by nifedipine treatment. AB - The effect of nifedipine (a calcium channel blocker) on the course of P. berghei infection was examined. It was observed that mice receiving a daily dose of 0.015 mg/kg of nifedipine had significantly shorter prepatent, patent and survival periods as compared to untreated P. berghei-infected animals (p < 0.001). This shows that the calcium channel blockers, in addition to possessing the property of reversing drug resistance during combined therapy with chloroquine, may also alter the pathophysiology of malaria infection. The decreased resistance of the host to the invading parasite suggests that the effect of CCB on the host parasite interaction in human malaria needs to be investigated further before CCB can be used in combination with chloroquine for the treatment of chloroquine resistant malaria or for chemoprophylaxis. PMID- 8240786 TI - An ultrastructural study of HIV-infected human dendritic cells and monocytes/macrophages. AB - The ultrastructure of dendritic cells (DC) isolated from HIV-negative blood donors revealed three morphologically distinct cell types: type I had an irregularly shaped nucleus with a high content of heterochromatin and numerous short pseudopodia; type II possessed a smoother boundary, a "blast-like" nucleus and a few veil-like protrusions; and type III resembled veil-like cells (DC) in the lymph. HIV induced a productive virus infection in type II DC with budding of virus solely from the cell surface. HIV was observed in cytoplasmic vacuoles of type III DC, but no budding of virus was observed from these cells. HIV was not observed in type I DC. Isolated monocytes were cultured for 8-15 days in order for these cells to differentiate into macrophages. Cultured monocytes and macrophages became highly vacuolized when infected with HIV strains and productive virus infection was localized in intracytoplasmic vacuoles. Budding from the cell surface was rarely observed. When monocytes were co-cultured with CD4(+)-infected lymphocytes, specialized electron-dense contact zones were observed in monocyte-like cells. Virus particles were found outside some of these contact zones, indicating that the zones may play a role in the penetration and transfer of virus from cell to cell. PMID- 8240787 TI - Multiplex polymerase chain reaction for detection of genes for Staphylococcus aureus thermonuclease and methicillin resistance and correlation with oxacillin resistance. AB - A multiplex polymerase chain reaction (mPCR) was used for simultaneous amplification of the staphylococcal nuc gene, encoding the thermostable nuclease (TNase), and the mecA gene, encoding the penicillin-binding protein 2a which is associated with staphylococcal methicillin resistance. A total of 219 staphylococcal strains were tested and the mPCR data were compared with coagulase production and in vitro oxacillin susceptibility. The agreement was 100% for coagulase production and nuc amplification, and 97.7%, 96.8 and 97.3% for mecA amplification and oxacillin resistance tested with MIC determination, disk diffusion and agar screen methods, respectively. Discrepant results were due to non-S. aureus isolates with borderline MICs of oxacillin (1-8 micrograms/ml). In a pilot test the mPCR simultaneously amplified both genes of staphylococci in blood cultures. This mPCR is a rapid and reliable method for single-step identification of cultures of MRSA and may prove to be useful for direct application on clinical specimens. PMID- 8240788 TI - P53 protein expression in Hodgkin's disease. AB - P53 is an oncosuppressor gene which is located on chromosome 17. Mutations of the p53 gene are closely associated with malignant transformation under in vitro conditions and are the most common genetic alteration in human malignancy. Unlike normal p53 protein which is unstable and usually cannot be detected by immunohistology, mutated p53 shows a decreased cell turnover rate and overexpression as compared with the wild-type protein. In this study a panel of four anti-p53 antibodies (PAb240, PAb421, PAb1801 and DO7) was applied to 52 cases of Hodgkin's disease: three cases of nodular lymphocytic predominance (LP), 33 cases of nodular sclerosis (NS), and 16 cases of mixed cellularity (MC). The results show that 53 protein is present in the Hodgkin's- and Reed-Sternberg cells in 82% of NS and 94% of MC, but not in nodular LP. It is suggested that mutations of the p53 gene and loss of normal p53 function are frequent in Hodgkin's disease and may be implicated in the pathogenesis of this disease. PMID- 8240789 TI - Cytokine plasma levels and lymphocyte subsets in patients with newly diagnosed insulin-dependent (type 1) diabetes mellitus before and following initial insulin treatment. AB - Changes in the plasma concentrations of interleukin-1 beta (IL-1 beta), tumour necrosis factor alpha (TNF alpha), interleukin 2 (IL-2), and lymphocyte subsets were investigated in 19 persons with newly diagnosed (type 1) insulin-dependent diabetes mellitus (IDDM) from admission to hospital prior to insulin treatment and following 1 week and 1 month of treatment. Furthermore, the cytokines were measured 16-28 months after the presentation of IDDM. The mean TNF alpha values were all within the normal range, but demonstrated a slight increase in all the samples taken (Friedman 0.06). The mean plasma IL-1 beta value was initially at the upper normal limit, but gradually increased significantly from admission to hospital to 1 week, 1 month, and 16-28 months afterwards (Friedman 0.031). No IL 2 activity was detectable in the majority of the samples. No significant changes in total leukocyte and lymphocyte counts were found. The lymphocyte subsets (CD5+, CD8+, CD4+, CD16+, CD20+, HLA-DR+) did not show any significant changes from admission to after the start of insulin treatment. It is concluded that the gradual increase in IL-1 beta and TNF alpha plasma levels may reflect an ongoing autoimmune inflammatory reaction at the onset of IDDM. PMID- 8240790 TI - Antibodies to Pneumocystis carinii in Danish blood donors and AIDS patients with and without Pneumocystis carinii pneumonia. AB - Sera from 1) 8 AIDS patients with Pneumocystitis carinii pneumonia, 2) 13 HIV antibody-positive patients, 9 with AIDS and 4 with AIDS-related symptoms, all 13 without Pneumocystis carinii pneumonia, and 3) 36 healthy male blood donors, all HIV antibody negative, were examined for antibodies against Pneumocystis carinii using the indirect immunofluorescence antibody technique. Seven (87%) out of eight sera from group 1, 12 (92%) out of 13 sera from group 2, and 35 (97%) out of 36 sera from group 3 were positive. There was no significant difference between group 1 and 2, and group 2 and 3. The difference between group 1 and group 3 was significant (p = 0.03). PMID- 8240791 TI - Serum reactivity to Chlamydia trachomatis and C. pneumoniae antigens in patients with documented infection and in healthy children by microimmunofluorescence and immunoblotting techniques. AB - Chlamydia trachomatis and C. pneumoniae are both important human pathogens. Antigenic cross-reactivity between the two species may complicate serologic diagnosis of infection with one or the other agent. In this study we examined sera of persons with chlamydia infections and of healthy children by microimmunofluorescence (MIF) against C. trachomatis L2 antigen and against C. pneumoniae TW-183 antigen to explore the degree of cross-reactivity found by these two methods. Likewise, the cross-reactivity seen by immunoblot with sera of rabbits immunized against one of the antigens when tested on the other was examined. While among healthy children stratified by age, MIF seropositivity to C. pneumoniae TW-183 increased with age, no such trend was observed with respect to seropositivity to C. trachomatis L2 antigens, and 81% of children seropositive to TW-183 did not react on L2 antigen. Moreover, 27% of those positive on L2 antigen were negative on TW-183. Immunoblot analysis showed much greater antibody cross-reactivity than that detected by MIF. The immunoblot cross-reactivity was probably not attributable solely to double infection since sera of rabbits immunized to only one species of chlamydia reacted strongly with both chlamydiae in immunoblot analysis. The data presented need to be taken into account in the development of serologic tests based on a small number of antigens or on partially denatured antigens. PMID- 8240792 TI - Antibiotic sensitivity of the Bacteroides fragilis group in Denmark. Danish Study Group. AB - Resistance rates to 9 antibiotics were determined for 370 isolates of the Bacteroides fragilis group submitted during a 5-month period in 1990-91 from 6 laboratories in Denmark. There was no resistance to imipenem (MIC > 8 mg/l) or metronidazole (MIC > 16 mg/l), while resistance to ampicillin was the rule (97% at > 4 mg/l) and to ceftriaxone rather common (19% at > 32 mg/l). Resistance was uncommon to cefoxitin (0.5% at > 32 mg/l), cefotaxime (6% at > 32 mg/l), piperacillin (7% at > 64 mg/l), ampicillin/sulbactam (1% at > 16 mg/l), and clindamycin (3% at > 4 mg/l). Two hundred and thirty-nine strains were Bacteroides fragilis. The 131 other strains were generally more resistant than the Bacteroides fragilis strains. For 5 antibiotics the Bacteroides fragilis had a significantly lower MIC in western than in eastern laboratories. For the group with other strains there was no significant difference. PMID- 8240793 TI - Pseudomonas aeruginosa bacteraemia detected with a new blood culture system Colorbact: a note of caution. AB - Detection of Pseudomonas aeruginosa bacteraemia by the newly introduced Colorbact blood culture system (Statens Seruminstitut, Copenhagen) was ascertained in a Danish regional department of clinical microbiology. Ten of sixteen bacteraemias (63%, 95% CL: 35-85%) were detected within 24 h; the 48 h figure was 81% (95% CL: 54-96%). The initial bacteriological diagnosis was confounded, however, by the observation of non-motile rods or atypical motility in 8 episodes, including 4 of the 10 episodes with a positive direct microscopy during the first 24 h. This may be an untoward effect of vigorous agitation. In two episodes P. aeruginosa was isolated only from the anaerobic culture bottle, whilst in two other episodes P. aeruginosa grew in aerobic and anaerobic bottles. This observation may reflect a chance distribution of viable bacteria among the bottles in the Colorbact set. PMID- 8240794 TI - The Wellcome Prize Lecture. A map of auditory space in the mammalian brain: neural computation and development. AB - The experiments described in this review have demonstrated that the SC contains a two-dimensional map of auditory space, which is synthesized within the brain using a combination of monaural and binaural localization cues. There is also an adaptive fusion of auditory and visual space in this midbrain nucleus, providing for a common access to the motor pathways that control orientation behaviour. This necessitates a highly plastic relationship between the visual and auditory systems, both during postnatal development and in adult life. Because of the independent mobility of difference sense organs, gating mechanisms are incorporated into the auditory representation to provide up-to-date information about the spatial orientation of the eyes and ears. The SC therefore provides a valuable model system for studying a number of important issues in brain function, including the neural coding of sound location, the co-ordination of spatial information between different sensory systems, and the integration of sensory signals with motor outputs. PMID- 8240795 TI - Potassium loss from skeletal muscle during exercise in man: a radioisotope study. AB - Muscle potassium (K+) content decreases during exercise. Previous studies, in humans, have used measurements of arteriovenous plasma potassium concentration differences (AV delta[K+]) and/or muscle biopsy to measure the loss of muscle K+ during exercise. In the current study a non-invasive method was developed to measure skeletal muscle K+ before and after exercise, using an isotope of K+, potassium-43 (43K+). Twelve subjects performed single-leg extension exercise for 2 h at 50% of their maximum predicted heart rate. The level of radioactivity from the quadriceps femoris was determined before exercise and during two periods post exercise. After correction for counts arising outside the exercised muscle, we estimate a decrease in muscle K+ content of 3.2 +/- 1.55% (mean +/- S.E.M.) following exercise. The muscle K+ was not restored following 75 min of recovery. The decrease in muscle K+ following exercise in our study is considerably less than that suggested by previous studies using AV delta[K+] measurements but not so dissimilar from results obtained using muscle biopsy. We conclude that a small but significant loss of K+ occurs following prolonged dynamic exercise, and that complete recovery of muscle K+ is slow. PMID- 8240796 TI - Rhythmicity associated with a high degree of short-term synchrony of motor unit discharge in man. AB - A rhythmic modulation of motor unit discharge at 17-24 Hz has been observed in the tibialis anterior muscle of subjects who display a high degree of short-term synchrony. Motor unit discharges were recorded both from extensor digitorum communis in the arm and tibialis anterior in the leg during voluntary isometric contractions. Time domain analysis of motor unit discharges in both muscles revealed a prominent central peak in the cross-correlograms that was interpreted as short-term synchrony. In tibialis anterior, but not in extensor digitorum communis, the primary, central peak was accompanied by secondary peaks. These were symmetrically or unilaterally present at an average time lag of 52 ms (range 37-68 ms) with respect to the primary peak. In both muscles other peaks occurred at time lags equivalent to the mean interspike interval of motor unit discharge (range 80-150 ms). In all subjects frequency domain analysis of the discharges of pairs of motor units revealed prominent peaks of coherence in the range 17-24 Hz. The actual frequency matched closely the reciprocal of the time lag interval between primary and secondary peaks of the cross-correlogram of the same motor units. Coherence was less evident for motor unit discharges in extensor digitorum communis and no prominent peaks occurred in the range 17-24 Hz. We propose that the secondary peaks and the rhythmicity may involve recurrent inhibition activated by the synchronous discharge of motor units. The presence of this pattern of discharge in tibilias anterior but not extensor digitorum communis would fit with recent animal and human work suggesting that recurrent inhibition is poorly represented in certain distal muscles of the forelimb. PMID- 8240797 TI - Neural modulation of bovine mesenteric lymph node contraction. AB - Spontaneous isometric contractions and field-evoked responses were recorded in vitro from strips of bovine mesenteric lymph node capsule. Field stimulation (0.3 ms pulses, 60 V nominal, 2 min trains) at frequencies > or = 2 Hz evoked a frequency-dependent increase in baseline tension which was maximum at frequencies > or = 16 Hz. Evoked contractions were significantly reduced (P = 0.02) by the neurotoxin tetrodotoxin (10(-6) M) although they were unaffected by the alpha adrenoceptor antagonists phentolamine, rauwolscine and prazosin (3 x 10(-6) M). Similarly, responses were unaffected by the beta-adrenoceptor antagonist propranolol (10(-6) M), the cholinergic antagonist atropine (10(-6) M) or the uptake blocker cocaine (10(-6) M). Field-evoked contraction was also unaffected by a 30 min exposure to alpha,beta-methylene ATP (10(-6) M). The results suggest that bovine lymph node capsular smooth muscle is innervated by excitatory nerves which are non-adrenergic, non-cholinergic and non-purinergic in nature. PMID- 8240798 TI - Periodic fluctuations in pancreatic secretion and duodenal motility investigated in neonatal calves. AB - To clarify the relative timing of clinical changes in duodenal motility and pancreatic secretion in newborn calves, we recorded duodenal electrical and mechanical activity and analysed pancreatic secretion and migrating myoelectric complex (MMC). In eight calves integrated recordings were derived from sites near the duodenal bulb and pancreatic accessory duct orifice, and pancreatic juice was sampled after an overnight fast, after a feed, and during reversible cold vagal blockade. Peak secretion coincided with duodenal irregular spiking activity and the nadir with absence of spiking. Feeding elicited electrical and mechanical hyperactivity in the duodenum, dissipated the MMC temporarily, and dramatically increased the juice volume and bicarbonate and protein outputs. Periodic fluctuations in secretion started shortly after a feed, as did recovery of the duodenal myoelectric complexes. Cold vagal blockade reversibly disrupted the synchronous changes in duodenal motility and pancreatic secretory activity, though the close association was not totally obliterated. In milk-fed calves interdigestive pancreatic secretion apparently rises and falls in phase with migrating myoelectric complexes of the proximal duodenum and the vagus is largely, though not exclusively, responsible for co-ordinating these changes. PMID- 8240799 TI - Volume-sensitive KCl co-transport and taurine fluxes in horse red blood cells. AB - Potassium (using 86Rb+ as a tracer), amino acid and taurine fluxes were measured in horse red blood cells (RBCs). No volume-sensitive component of alanine and glycine transport was observed, and although volume-sensitive taurine fluxes were observed in most animals, their absolute magnitudes were small. K+ fluxes, however, were shown to be particularly volume sensitive; they were stimulated by cell swelling and inhibited by cell shrinkage. Sizeable fluxes were present at normal cell volumes. The volume-sensitive K+ flux was Cl- dependent and was abolished by Cl- replacement with methylsulphate. The Cl(-)-dependent K+ fluxes in horse red blood cells were stimulated by lowering in external pH to 6.9 and by treatment with the sulphydryl-reacting agent, N-ethylmaleimide. They were inhibited by the potent K(+)-Cl- co-transport inhibitor, DIOA, ([(dihydroindenyl)oxy]alkanoic acid) but were insensitive to the Na(+)-K(+)-Cl- co-transport inhibitors, frusemide and bumetanide. A Cl- channel inhibitor, 5 nitro-2-(phenylpropyl-amino)-benzoate (NPPB), produced partial inhibition. These results suggest that regulatory volume decrease in horse red blood cells is achieved predominantly by volume-sensitive K+ efflux mediated via a K(+)-Cl- co transport system with similar properties to those observed in the red blood cells of other species. The significance of these findings and their rheological consequences are discussed. PMID- 8240800 TI - Insulin and intestinal epithelial cell proliferation. AB - The effects of modification of the levels of endogenous plasma insulin on intestinal epithelial cell proliferation were investigated in rats maintained by total parenteral nutrition (TPN) using either a TPN diet that was high in glucose, or a low glucose (hypocaloric) TPN diet and in orally fed rats (normal and sham operated). The rats were put on their respective treatments for 7 days and were then injected with vincristine sulphate to arrest cells as they entered metaphase. Intestinal tissue was fixed for the determination of crypt cell production rate (CCPR) and blood plasma was taken for hormone radioimmunoassay. Sham operation had no effect on either plasma hormone levels or on CCPR. The standard TPN diet was associated with significantly reduced levels of gastrin, peptide YY and enteroglucagon. Gastrin and PYY were further reduced by the hypocaloric diet. However, plasma insulin was much increased in the TPN but not in the hypocaloric group. Small intestinal CCPR was reduced in the TPN groups, but no difference was observed between the standard and the low calorie TPN diets. The lack of difference in CCPR between the two TPN diets despite the massive elevation of plasma insulin strongly suggests that insulin does not have a substantial role in the control of gastrointestinal epithelial cell renewal. PMID- 8240801 TI - Sound stimulation increases the cerebral glucose:oxygen quotient in the fetal sheep. AB - We sought to determine the cerebral metabolic fate of glucose during sound stimulation in near-term fetal sheep. Blood oxygen content was significantly decreased by 6-7% during sound stimulation. The cerebral glucose:oxygen quotient increased from 0.88 +/- 0.08 to 1.13 +/- 0.12 (mean +/- S.E.M., P < 0.01). This is consistent with the phenomenon of stimulated glycolysis which has been shown to increase brain lactate concentration in other models. PMID- 8240802 TI - Measurements of intracellular Ca2+ in cultured human myometrial smooth muscle cells bathed in low Na+ solutions. AB - In cultured human myometrial smooth muscle cells, removal of external Na+ activates repetitive increases in intracellular Ca2+ ([Ca2+]i). The Ca2+ transients persist in isotonic K+ solutions which suggests that the activity does not arise from regenerative changes in membrane potential. In nominally Ca(2+) free solution, the activity disappears after a few cycles suggesting the involvement of internal stores. On Na+ removal, a background influx of Ca2+ may be responsible for activating the cyclical release of Ca2+ from internal stores. The absence of any effect of caffeine on resting [Ca2+]i suggests that the classical cardiac type of Ca(2+)-induced-Ca(2+)-release mechanism is not operating in these cells. PMID- 8240803 TI - Bombesin and gastrin-releasing peptide stimulate electrogenic ion transport in cultured human endometrial epithelial cell layers. AB - Electrogenic ion transport across human endometrial epithelial cells grown as polarized monolayers on permeable supports was measured as an inward short circuit current (Isc; 16.2 +/- 1.1 microA/cm2). Bombesin (10(-7) M) and human gastrin-releasing peptide (GRP; 10(-7) M) caused transient enhancement of this Isc. These effects were largely restricted to the basolateral surface of the cells; responses to apical peptide were modest in comparison with those to basolateral peptide. GRP and other bombesin-related peptides may have a role in regulation of endometrial epithelial ion transport in vivo and thereby influence the intra-uterine environment. PMID- 8240804 TI - Rab proteins and the road maps for intracellular transport. PMID- 8240805 TI - Gradual loss of synaptic cartels precedes axon withdrawal at developing neuromuscular junctions. AB - We have studied the spatial deployment of synapses arising from different axons that converge on the same developing neuromuscular junctions. Labeling the competing synaptic "cartels" with different dyes in mouse muscle showed that, perinatally, each axon adds similar terminal areas, whereas later, areas occupied by the competing cartels diverged by gradual elimination of one axon's synapses and ongoing addition of synaptic area by the other. Activity-dependent labeling of synapses capable of vesicle recycling in snake muscle also revealed a gradual change in territories occupied by competing inputs, implying that an axon maintained some functional synapses even as others in its cartel were being eliminated. Thus the process of synapse elimination is gradual, with loss of one viable synapse after another, until an axon is left with no synaptic territory and withdraws. PMID- 8240806 TI - Compositional changes of AP-1 DNA-binding proteins are regulated by light in a mammalian circadian clock. AB - Recent reports have shown that the nuclear phosphoprotein Fos is induced by light in a mammalian circadian clock, the suprachiasmatic nucleus. To learn how light and circadian phase affect the binding of Fos to DNA, we analyzed the photic and temporal regulation of immunoreactive Jun protein expression and AP-1 DNA-binding activity in the rat suprachiasmatic nucleus. Immunohistochemistry and gel mobility shift assays suggest that AP-1 activity during the night and after a light pulse consists of constant, as well as variable, protein components; JunD could be identified as a constituent of both dark- and light-activated binding complexes, whereas binding by JunB and Fos could be implicated only after photic stimulation. Since JunD or JunB could be colocalized with Fos in individual suprachiasmatic nucleus cell nuclei, light may be acting in at least some suprachiasmatic nucleus cells by altering AP-1 protein composition rather than binding site occupancy. PMID- 8240807 TI - Histamine potentiates N-methyl-D-aspartate responses in acutely isolated hippocampal neurons. AB - N-methyl-D-aspartate (NMDA)-evoked currents were recorded from acutely isolated rat hippocampal neurons, using the whole-cell patch-clamp technique and a rapid perfusion system. Histamine, at concentrations from 0.5 to 100 microM, reversibly enhanced NMDA currents by up to 50%. The effect cannot be ascribed to activation of the known histamine receptors (H1, H2, H3) but is occluded by spermine. These results suggest an interaction of histamine with the polyamine-binding site on the NMDA receptor complex. This modulatory action could allow the histaminergic system to determine time and loci of NMDA receptor-mediated events, such as memory formation according to behavioral state. PMID- 8240808 TI - Ca2+ is an obligatory intermediate in the excitation cascade of limulus photoreceptors. AB - We have investigated the role of Ca2+ in the excitation of Limulus photoreceptors by intracellular injection of the Ca2+ buffer, 5,5'-dibromo-BAPTA. Buffer with free Ca2+ of 0.5 or 5 microM slowed the rising edge of the light response over 100-fold and greatly reduced both the transient and plateau phases of the light response, as expected if Ca2+ elevation is necessary for all phases of excitation. Injection of buffers with free Ca2+ of 5 or 45 microM, levels normally reached during light, evoked sustained inward current as expected if Ca2+ is sufficient for excitation. The transduction cascade appears due to a single pathway that sequentially involves 1,4,5-trisphosphate inositol, Ca2+, and cyclic GMP. PMID- 8240809 TI - Allosteric modulation of the NMDA receptor by dihydrolipoic and lipoic acid in rat cortical neurons in vitro. AB - The mitochondrial cofactor dihydrolipoic acid (DHLA) was observed to potentiate N methyl-D-aspartate (NMDA), but not non-NMDA, receptor-mediated whole-cell responses in cultured neurons. This potentiation was readily reversed by the oxidizing agent 5,5'-dithio-bis-(2-nitro-benzoic acid) (DTNB). DHLA was unable to increase NMDA responses previously potentiated by dithiothreitol, nor did it have an effect on NMDA receptors alkylated with N-ethylmaleimide. Single-channel recordings revealed that DHLA produced an increase in NMDA channel open frequency, with no change in single-channel conductance or open time. In contrast, lipoic acid reversed the potentiation of NMDA-evoked responses produced by dithiothreitol and had no effect on NMDA receptors previously oxidized by DTNB. DHLA and lipoic acid are pervasively found substances that readily permeate cellular membranes and thus may influence NMDA receptor activity in vivo by modifying its redox site. PMID- 8240810 TI - The Wnt-1 proto-oncogene induces changes in morphology, gene expression, and growth factor responsiveness in PC12 cells. AB - The product of the Wnt-1 proto-oncogene is a secreted glycoprotein that is normally produced in regions of the embryonic neural tube. We show here that expression of mouse Wnt-1 cDNA in the rat PC12 pheochromocytoma cell line causes a dramatic conversion from a round to a flat cell morphology. In addition, PC12 cells expressing Wnt-1 (PC12/Wnt-1) fail to extend neurites after treatment with NGF, despite the presence and activation of high affinity NGF receptors encoded by the trk gene and the induction of early response genes. Furthermore, PC12/Wnt 1 cells fail to express several neuron- and chromaffin-specific genes, indicating that PC12/Wnt-1 cells have assumed a new phenotype. Although NGF and FGF utilize similar signal transduction pathways in PC12 cells, only FGF is capable of inducing a morphological response and synthesis of transin mRNA in PC12/Wnt-1 cells. PMID- 8240811 TI - Neighboring cerebellar Purkinje cells communicate via retrograde inhibition of common presynaptic interneurons. AB - Paired tight-seal whole-cell recordings were obtained from neighboring Purkinje cells in cerebellar slices. Under voltage clamp, spontaneous inhibitory postsynaptic currents resulted from the activity of GABAergic interneurons, stellate and basket cells. Up to 80% of inhibitory postsynaptic currents in paired recordings were in register. This correlation was not affected by antagonists of glutamate receptors, faded with distance, and was abolished by tetrodotoxin. Earlier work showed that voltage-gated Ca2+ entry into a Purkinje cell elicits a transient presynaptic inhibition of inhibitory postsynaptic currents. It is now shown that this inhibition is not restricted to the stimulated cell, but that it is transmitted to its neighbors. The results indicate that Purkinje cells exchange information by an unconventional mechanism involving retrograde control of inhibitory synapses. PMID- 8240812 TI - Cloning and pharmacological characterization of a rat mu opioid receptor. AB - We have isolated a rat cDNA clone that displays 75% amino acid homology with the mouse delta and rat kappa opioid receptors. The cDNA (designated pRMuR-12) encodes a protein of 398 amino acids comprising, in part, seven hydrophobic domains similar to those described for other G protein-linked receptors. Data from binding assays conducted with COS-1 cells transiently transfected with a CMV mammalian expression vector containing the full coding region of pRMuR-12 demonstrated mu receptor selectivity. In situ hybridization mRNA analysis revealed an mRNA distribution in rat brain that corresponds well to the distribution of binding sites labeled with mu-selective ligands. Based upon these observations, we conclude that pRMuR-12 encodes a mu opioid receptor. PMID- 8240813 TI - Expression of the sodium channel beta 1 subunit in rat skeletal muscle is selectively associated with the tetrodotoxin-sensitive alpha subunit isoform. AB - Transcripts homologous to the rat brain sodium channel beta subunit (beta 1) are prominently expressed in both innervated and denervated adult skeletal muscle and in heart, but not in neonatal skeletal or cardiac muscle. Regulation of beta 1 mRNA expression closely parallels that of SkM1 alpha during development, after denervation in adult muscle, and in primary muscle culture, but does not follow SkM2 expression under any condition examined. In oocytes, beta 1 interacts functionally with SkM1 to modulate the abnormally slow inactivation kinetics observed with this alpha subunit expressed alone. We conclude that a common beta 1 subunit is expressed in skeletal muscle, heart, and brain and that in skeletal muscle, this subunit is specifically associated with the SkM1, rather than the SkM2, sodium channel isoform. PMID- 8240814 TI - Transient period of correlated bursting activity during development of the mammalian retina. AB - The refinement of early connections in the visual pathway requires electrical activity in the retina before the onset of vision. Using a multielectrode array, we have shown that the spontaneous activity of cells in the neonatal ferret retina is correlated by patterns of periodically generated traveling waves. Here, we examine developmental changes in the characteristics of the waves and show that retinal ganglion cells participate in these patterns of activity, which are seen during the same period as synaptic modification in the lateral geniculate nucleus; that the waves subside gradually as the connectivity in the lateral geniculate nucleus stabilizes; and that their spatial structure allows for refinement of the retinotopic map, as well as for eye-specific segregation in the lateral geniculate nucleus. PMID- 8240815 TI - A GTPase-accelerating factor for transducin, distinct from its effector cGMP phosphodiesterase, in rod outer segment membranes. AB - Hydrolysis of GTP by the photoreceptor G protein transducin (Gt alpha) was found to occur with kinetics identical to the inactivation of its effector cGMP phosphodiesterase (PDE), but was too slow (tens of seconds) in dilute rod outer segment (ROS) suspensions to account for subsecond recovery of the light response. Raising the concentration of ROS membranes increased the rates of GTP hydrolysis and PDE inactivation in parallel as much as 6-fold. Holo-PDE and its gamma subunit had weak effects on GTPase kinetics (< 1.6-fold and < 1.3-fold, respectively). ROS membranes stripped of PDE retained approximately 90% of a GTPase accelerating activity that was protease sensitive, indicating that they contain a GTPase-accelerating factor distinct from PDE. PMID- 8240816 TI - bFGF regulates the proliferative fate of unipotent (neuronal) and bipotent (neuronal/astroglial) EGF-generated CNS progenitor cells. AB - In cultures of embryonic and adult mouse striatum, we previously demonstrated that EGF induces the proliferation of putative stem cells, which give rise to spheres of undifferentiated cells that can generate neurons and astrocytes. We report here that the spheres of undifferentiated cells contain mRNA and protein for the FGF receptor (FGFR1). Indirect immunocytochemistry demonstrated that many of the cells within the EGF-generated spheres were immunoreactive for FGFR1. Exogenous application of bFGF to the EGF-generated cells induced the proliferation of two progenitor cell types. The first, a bipotent progenitor cell, gave rise to cells with the antigenic and morphological properties of neurons and astrocytes; the other gave rise to cells with neuronal characteristics only. bFGF-generated cells with neuronal morphology exhibited electrophysiological properties indicative of immature central neurons. These results support the hypothesis that sequential actions of growth factors play a role in regulating the generation of neurons and astrocytes in the developing CNS. PMID- 8240817 TI - A peptide segment critical for sodium channel inactivation functions as an inactivation gate in a potassium channel. AB - The short cytoplasmic peptide segment connecting domains III and IV of voltage gated sodium channels (III-IV linker) is essential for fast inactivation. To test the functional similarity between the III-IV linker and the potassium channel inactivation particle, we attached the III-IV linker to the amino terminus of a noninactivating potassium channel. This chimeric channel inactivated rapidly and displayed biophysical properties similar to Shaker A-type potassium channels. Recovery from inactivation in the chimeric channels was accelerated by high external potassium, consistent with the idea that potassium ions passing through the channel displaced the III-IV linker inactivation particle. A mutation that completely abolishes fast inactivation in rat brain sodium channels also completely abolished inactivation in the chimera. These results demonstrate that the sodium channel III-IV linker can function as a fast inactivation gate and suggest a functional relationship between the fast inactivation processes of sodium and potassium channels. PMID- 8240818 TI - Appearance of target-specific guidance information for regenerating axons after CNS lesions. AB - During development of the vertebrate visual system, an orderly projection of ganglion cells from the retina onto the superior colliculus (SC) is established. Mechanisms that might govern this process include the coordinated action of guidance and corresponding receptor molecules that are specifically distributed on the axons and their targets. In birds and mammals, information for axonal guidance and targeting appears to be confined to the time when the retinocollicular projection is being formed. Here we show that putative guidance activities for temporal and nasal retinal axons, which are not detectable in the normal adult SC, appear after optic nerve transection in adult rats. Both embryonic and adult retinal axons are able to respond to these guiding cues, although the guidance activities detectable in the deafferented adult rat SC might be different from those found during development. These findings imply that it might be possible to reestablish an ordered projection after lesions in the adult mammalian visual system. PMID- 8240819 TI - Huntington's disease gene (IT15) is widely expressed in human and rat tissues. AB - Huntington's Disease (HD) is notable for selective neuronal vulnerability in the basal ganglia and cerebral cortex. We have investigated in human and rodent tissues the expression of the gene (IT15) whose mutation causes HD. IT15 is widely expressed, with highest levels of expression in brain, but also in lung, testis, ovary, and other tissues. Within the brain, expression is widespread with a neuronal pattern and is not enriched in the basal ganglia. Expression of IT15 is not reduced in the brain of HD patients when corrected for actin (though it is slightly decreased in the striatum when uncorrected, consistent with neuronal loss). Thus, the widespread distribution of IT15 expression does not correspond with the restricted distribution of neuropathologic changes in HD. We suggest that pathophysiology may relate to abnormal cell type-specific protein interactions of the HD protein. PMID- 8240820 TI - The structure and regulation of tight junctions. AB - Tight junctions create a paracellular barrier between both epithelial and endothelial cells. Recent advances have helped define their molecular composition and regulation. Studies in cultured cell lines provide new insights into how assembly and barrier properties may be controlled by signal transduction cascades. PMID- 8240821 TI - The analysis of neural recognition molecules: benefits and vicissitudes of functional knock-outs using antibodies and gene ablation. AB - Altering or removing neural recognition molecules by blocking antibodies or genetic deletion has led to a flurry of predictions concerning their function. Each experimental approach has advantages and disadvantages that have to be considered when interpreting the resulting phenotypes. PMID- 8240822 TI - Factors regulating the expression and function of calcium-independent cell adhesion molecules. AB - A combination of genetic and biochemical analysis is contributing to an understanding of the regulation and role of calcium-independent cell adhesion molecules. Recent progress ranges from the analysis of the control of promoter expression by homeobox genes to detailed analysis of the sites of homophilic and heterophilic interactions via mutagenesis strategies. PMID- 8240823 TI - The structural and functional analysis of cadherin calcium-dependent cell adhesion molecules. AB - During the past year considerable progress has been made in our understanding of cadherin structure and function. Recent research has concentrated on several aspects of the cell and molecular biology of cadherins, including genomic organization, cytoskeletal interactions, regulation of expression and function by post-translational modifications, differential expression during embryonic development, and the emerging role of cadherin misexpression and malfunction in pathogenesis. PMID- 8240824 TI - Cadherins in cancer: implications for invasion and metastasis. AB - In order to understand the mechanisms of metastasis, one must first determine how cancer cells detach from primary tumors. It is known that cadherins are major cell-cell adhesion molecules in tumors as well as in normal tissues. The perturbation of cadherin function causes temporal or permanent disaggregation of tumor cells and may thus promote the invasion and metastasis of such cells. PMID- 8240825 TI - Adhesion molecules in cancer: the role of integrins. AB - It has been known for some time that cell adhesion receptors, including members of the integrin family, play an important role in the biology of tumors. Until recently, most of the emphasis in this area of research has concerned the functioning of integrins as adhesive molecules in the invasive and metastatic behavior of malignant cells. Now it has become clear that integrins can function as true receptors capable of transducing signals to the cell interior. Tyrosine phosphorylation seems to be a key aspect of integrin-mediated signal transduction, and a new tyrosine kinase has been described that seems to be important in this process. Evidence is accumulating that integrin-mediated signals can induce gene expression and affect transit through the cell cycle. Thus, the role of integrins in cancer seems not only to involve cell adhesion events, but may also involve the regulation of tumor cell growth and differentiation. PMID- 8240826 TI - Integrin cytoplasmic domains: mediators of cytoskeletal linkages and extra- and intracellular initiated transmembrane signaling. AB - The integrin cytoplasmic domains are generally short amino acid sequences that are highly divergent among alpha-subunits and partially conserved among beta subunits. Interest in them has increased over the past few years as they have been shown to mediate several kinds of transmembrane signaling events as well as structural connections between the extracellular matrix and the cytoskeleton. PMID- 8240827 TI - Cell interactions in invertebrate development. AB - By utilizing both genetics and physical manipulations, studies on invertebrate embryos have provided substantial information on how cell interactions influence cell fate during development. The picture emerging from these studies suggest that while there may be varied and intricate patterns of cell interactions during the development of different embryos, the signaling molecules that convey positional information across cell membranes have been highly conserved. PMID- 8240828 TI - The coordinated regulation of heparan sulfate, syndecans and cell behavior. AB - Recent advances in our understanding of heparan sulfate proteoglycans at the cell periphery implicate them as important participants in cell surface signaling. Regulation of the expression of discrete core proteins and of the specificity inherent in their heparan sulfate chains is thus emerging as a critical means of controlling cell behavior. PMID- 8240829 TI - Glycosyltransferases as cell adhesion molecules. AB - Specific glycosyltransferases are believed to be present on the cell surface, where they may function as cell adhesion molecules by binding oligosaccharide substrates on adjacent cell surfaces or in the extracellular matrix. The best studied of these is beta 1,4-galactosyltransferase, which mediates sperm binding to the egg coat and selected cell interactions with the basal lamina. Molecular models have recently been presented to account for the expression of beta 1,4 galactosyltransferase on the cell surface, thus enabling one to alter its surface expression and to examine the consequences on fertilization and development. PMID- 8240830 TI - The laminin family. AB - During the preceding year significant progress has been made in our understanding of laminin structure and expression. Highlights include the identification of new subunit chains and molecular isoforms as well as new information on tissue specific gene expression, laminin self-assembly and interactions with other matrix components. PMID- 8240831 TI - New members of the collagen superfamily. AB - During the past year, several novel collagenous clones have been isolated from cDNA libraries and designated as new collagen types. Recent research has been concerned with describing these new collagens and their potential relationships to other previously described collagen types. PMID- 8240832 TI - Structural biochemistry and activation of matrix metalloproteases. AB - The degradation of the extracellular matrix is part of many pathological and physiological processes. Of the several proteases involved in extracellular matrix turnover, the plasmin/plasminogen activator system and the family of matrix metalloproteases have received the most attention. Recent investigations in the field of matrix metalloprotease biochemistry have focused on the functions of the various enzyme domains and their interactions with inhibitor domains. Research into physiological activation mechanisms has demonstrated a plasmin/plasminogen activator-metalloprotease cascade, as well as providing an initial characterization of cell surface associated metalloprotease activation. PMID- 8240833 TI - von Willebrand factor and fibrinogen. AB - Progress in understanding the structure and function of von Willebrand factor and fibrinogen has provided new insights into the mechanisms of hemostasis and thrombosis. This, in turn, has generated concepts that may become applicable to the diagnosis and treatment of occlusive vascular diseases and bleeding disorders, both congenital and acquired. PMID- 8240834 TI - Cell-to-cell contact and extracellular matrix. PMID- 8240835 TI - [Primary antiphospholipid syndrome in juvenile ischemic stroke]. AB - Results obtained in three patients with juvenile ischemic stroke and criteria of primary antiphospholipid syndrome is reported. These patients are selected out from a series of 12 patients with 18-mounts follow-up. Lupic anticoagulant and anticardiolipin antibodies were found in two of three patients and one patient show anticardiolipin antibodies with negative lupic anticoagulant. All others coagulation proteins examined (antithrombin III, plasminogen, protein C and protein S) were normal. We conclude that antiphospholipid antibodies are associated with increase risk of thrombosis. Therefore should be systematically investigated in juvenile ischemic stroke of unknown aetiology. PMID- 8240836 TI - [Efficacy of the proteic redistribution diet (PRD) in the antiparkinsonian effect of L-dopa]. AB - The efficacy of PRD was evaluated in 21 patients with Parkinson's disease who presented motor fluctuations or a bad quality "on" time despite treatment with L dopa or dopaminergic agonists. Eighteen of the 21 patients completed three months of treatment and only 14 (66.6%) were able to complete 12 months. The four patients who stopped treatment in this period did so because of subjective complaints despite obtaining good clinical response to PRD. Only three patients suspended the dietetic regime during first month of treatment. The 14 who completed the year of treatment showed a decrease in the "off" time (4.10 to 1.65: p < 0.01) and an improvement in motor response (14.89 to 10.39; p < 0.05) during the follow up period. Improvement appeared at the end of the second week of initiation of treatment. The diet was well tolerated with scarce complications, with only one patient presenting visual hallucinations. Other secondary effects were mild and did not require discontinuation of the diet. We conclude that proteic redistribution diet improves the "on" time in patients with Parkinson disease receiving L-dopa and delays the possibility of increasing the doses of L-dopa. The dietetic regimens with proteic redistribution seems a simple, useful and cheap approach without morbidity for the patients with Parkinson's disease showing a poor response or oscillation to L-dopa treatment. PMID- 8240837 TI - [Retinal periphlebitis in multiple sclerosis. A prospective study]. AB - Twenty-four patients with clinically defined multiple sclerosis were prospectively studied with the aim of establishing the frequency of retinal periphlebitis. In three cases (12.5%) retinal periphlebitis was observed. None of the patients with multiple sclerosis and retinal periphlebitis presented a severe form or progressive course of the disease; however, in one patient it caused complete unilateral amaurosis. Aggressive immunosuppressive treatment was effective in one case. Given the absence of myelin in the retina, the presence of retinal periphlebitis suggests the existence of a vascular mechanism in the pathogenesis of multiple sclerosis. PMID- 8240838 TI - [Parkinson's disease etiology: risk factors and protection]. PMID- 8240839 TI - [Restless legs syndrome and periodic movements during sleep]. PMID- 8240840 TI - [Magnetic resonance of the brain and Achilles tendon in cerebrotendinous xanthomatosis]. AB - We report a 31 year old patient with cerebrotendinous xanthomatosis who presented with dementia, tendinous xanthomas, cataracts, pyramidal involvement, cerebellar ataxia and peripheral neuropathy. Cerebral CT scan demonstrated diffuse hypodensity in the white matter in both cerebellar hemispheres while MR detected additional focal alterations in both frontal lobes. MR study of the Achilles tendon showed a diffuse enlargement of the tendon with multiple areas of hypersignals in T1 and T2 demonstrative of the lipid deposits, interpossed between the isosignal zones that may correspond to the inflammatory reaction secondary to the accumulation of cholesterol and cholestanol. PMID- 8240841 TI - [Neoplastic lumbosacral plexopathy and "hot foot"]. AB - Neoplastic lumbosacral plexopathy is a frequent and disabling complication in subjects with cancer. Its clinical presentation is characterized by pain, muscle weakness, sensory complaints in one or occasionally both limbs associated with the tumoral symptoms. The presence of autonomic symptoms is less frequent; one of these is the "hot and dry foot". We present two patients, one with prostatic cancer and the other with myxoid liposarcoma, who developed a lumbosacral plexopathy as a manifestation of the extension of the neoplastic process; in both cases there was in addition a clear difference in the temperature of the affected limb. Although infrequent, the "hot and dry foot" it constitutes an early sign of metastatic plexopathy which facilitates the differential diagnosis with preforaminal lumbosciatic radiculopathies. PMID- 8240842 TI - [Benign amaurosis fugax]. AB - In young patients episodes of amaurosis fugax usually correspond to processes not related with atherosclerosis of the extracranial vessels such as migraine, cardiac embolism, vasospasm, vasculitis, oral contraceptives or hematologic disease. In 11% of the cases, however, no cause can be found. These idiopathic cases generally have a benign course and their frequency decreases spontaneously allowing them to be described as benign amaurosis fugax. We describe four cases of amaurosis fugax in which no responsible cause was found despite extensive clinical, laboratory ultrasonographic and angiographic studies. The episodes progressively remitted. In these cases of benign amaurosis fugax the risk of a permanent deficit is exceptional and treatment is not necessary. PMID- 8240843 TI - [Neurologic complications induced by the treatment of the acute renal allograft rejection with the monoclonal antibody OKT3]. AB - The treatment of the acute renal allograft rejection with the monoclonal antibody orthoclone OKT3 produces both systemic and neurologic alterations. In a series of 21 patients with an acute renal allograft rejection treated with this monoclonal antibody, 20 with a renal allograft transplantation and one with a renal and pancreatic allograft transplantation, 29% referred headache associated with fever and vomiting, and 14.2% presented severe neurological alterations induced by the treatment. We stress the need to know these secondary effects to differentiate them from other central nervous system disorders, particularly those of infectious origin. PMID- 8240844 TI - [Thyrotoxic periodic paralysis]. PMID- 8240845 TI - [Hemicrania, headache, migraine?]. PMID- 8240846 TI - Factors in dentin bonding. Part II: A review of the morphology and physiology of human dentin. PMID- 8240847 TI - Porcelain laminate veneers: ten years later. Part I: Tooth preparation. PMID- 8240848 TI - Esthetic high-strength implant abutments. Part II. PMID- 8240850 TI - The difficult patient stress syndrome: Part I. PMID- 8240849 TI - Romancing the beautiful silver maiden: an allegorical love story. PMID- 8240851 TI - A study of self poisoning with oral salbutamol--laboratory and clinical features. AB - 1. The recent increase in asthma mortality coupled with reports of fatal asthma associated with beta-2-agonist therapy, has stimulated interest in the plasma concentrations of beta-2-agonists that produce systemic toxicity. 2. We prospectively studied 17 patients (9 male), mean age 23 years (range 2-72), who attended the emergency departments of hospitals throughout the United Kingdom having recently ingested an overdose of salbutamol. 3. Clinical, laboratory, ECG data, plasma and urine samples were obtained from each patient. Plasma was assayed for salbutamol concentration using a high performance TLC photodensitometric method. 4. The mean (+/- s.d.) salbutamol dose reported to have been ingested was 89(+83)mg and the mean plasma salbutamol concentration was 166 (range 18-449) ng ml-1. The mean plasma potassium was 2.9 (s.d. +/- 0.6) mM (n = 16). None of the patients in this study developed serious cardiac dysrrhythmias. 5. There were significant correlations between the plasma salbutamol concentration and plasma potassium concentration (r = -0.85; P < 0.00005) and between plasma salbutamol concentration and pulse rate (r = 0.66; P < 0.005). 6. We conclude that in these patients, without respiratory decompensation, suprapharmacological plasma concentrations of salbutamol were tolerated without serious cardiac arrhythmias or any fatalities. PMID- 8240852 TI - Collagen fibrillar network in estrous and hCG-stimulated rabbit uterus: a SEM study after NaOH maceration. AB - The three-dimensional (3-D) organization of the collagen fibrillar network in rabbit uterus was studied by cell maceration/scanning electron microscopy (SEM) method, both during estrus and after stimulation with 100 I. U. of human chorionic gonadotropin (hCG). The cellular elements of glutaraldehyde-fixed tissue were effectively removed with 2N-NaOH at 25 degrees C for 5-7 days, followed by rinsing in distilled water. Correlative light microscopy (LM) and transmission electron microscopy (TEM) observations were also made in order to better clarify the natural location and distribution of collagen structures in the uterine layers. In estrous and ovulatory rabbits the superficial compartment of the endometrial stroma was composed of diffusely organized individual collagen fibrils and thin bundles of fibrils, forming specific supporting structures around the uterine glands. The deeper stroma showed a more condensed lamellar arrangement, particularly in the central part of the endometrial folds (plicae). Two distinct collagen networks, detected in the circular and longitudinal muscle layers of the myometrium, represented plexuses of anastomosing fine sheaths around the individual smooth muscle cells. A condensation of the fibrillar framework was observed around the large blood vessels between these two layers. Little alteration in uterine fibrillar network could be noticed in ovulatory animals 12 h after stimulation with hCG. Marked changes indicating collagen reorganization occurred in progestational animals 7.5 days after hCG administration. In this case, the collagen fibrillar network of the endometrium showed an irregular pattern consisting of slender elongations surrounding the labyrinthine spaces of the epithelium and corresponding to numerous, extremely attenuated plicae. Such a rearrangement of fibrils is very likely a response to hormonal stimulation of the uterus. To further identify the location of the blood vessels in the uterine collagen skeleton, vascular casts were also prepared. The collagen fibrillar network not only provides a mechanical support for the structural components of the uterus but also creates a favourable milieu for their activities. PMID- 8240853 TI - Microcirculatory patterns in adult rat cerebral hypophysis: a scanning electron microscope study of replicated specimens. AB - The blood vascular bed of the cerebral hypophysis in the adult rat was replicated completely or incompletely by arterial injection of different amounts of methacrylate resin, to be observed with a scanning electron microscope. Complete replication confirmed our previous findings (Murakami et al., 1987) on the distribution and structure of the vascular beds in and around the hypophysis of the rat. One long major and several minor portal routes (vide infra) were reproduced sufficiently together with the systemic veins of the posterior lobe. Incomplete replication demonstrated that resin flows: 1) via the long portal vessels from the median eminence and neural stalk to the anterior lobe; 2) via the accessory long portal vessels from the subependyma to the anterior lobe; 3) via the short portal vessels from the posterior lobe to the anterior lobe; 4) via the neuro-intermedial portal vessels from the posterior lobe to the intermediate lobe; 5) via the intermedio-distal portal vessels from the intermediate lobe to the anterior lobe; and 6) via the tuberal portal vessels from the tuberal lobe to the anterior lobe. Incomplete replication also demonstrated that resin in the median eminence and neural stalk is drained preferentially into the anterior lobe via the long portal vessels, and that resin in the posterior lobe is drained mainly into the systemic veins. We were unable to demonstrate a retrograde resin flow from the anterior lobe to the median eminence, subependyma, neural stalk, intermediate lobe and posterior lobe, nor an ascending resin flow from the posterior lobe to the median eminence and subependyma. Also failing to be noted were an ascending resin flow from the hypophysis to the hypothalamus and a descending resin flow from hypothalamus to the hypophysis. PMID- 8240854 TI - Heterogeneity of motilin-immunoreactive cells in the duodenum and pyloric region of several avian species. AB - Immunohistochemical characterizations of motilin-immunoreactive cells were examined in gastric and duodenal mucosae of nine species of birds from seven orders using five different region-specific motilin antisera. Motilin immunoreactive cells appeared as open-type cells in the mucosal epithelium and showed varying immunoreactivities to antisera used in all the birds examined except for the cormorant and penguin, which did not show any kinds of immunoreactivity to motilin. Motilin-immunoreactive cells of the emu duodenum were detected by all the motilin-antisera used. The present results suggest that there is a wide range of heterogeneity between motilin molecules among avian species, or perhaps alternatively the existence of a family of motilin-like peptide. Furthermore, the present results should prove useful for a molecular biological study on the evolution of avian motilin. PMID- 8240855 TI - Immunocytochemical localization of basic fibroblast growth factor in the rat pituitary gland. AB - Immunohistochemical localization of basic fibroblast growth factor (bFGF) was studied in the rat pituitary gland at light and electron microscopic levels using a specific monoclonal antibody. In the anterior lobe, bFGF-immunoreactivity was found mostly in a cell population without secretory granules, which was identified as the folliculo-stellate (FS) cell by its ultra-structure and expression of S-100 protein. The subcellular localization of the immunoreactivity in FS cell was strictly nuclear. Besides the FS cell, a minor cell population containing small granules of 60-100 nm diameter, which seemed to represent immature endocrine cells, was immunostained in the nuclei with the anti-bFGF antibody. In the intermediate lobe, a subpopulation of the S-100 protein-positive cells lining the pituitary cleft was immunoreactive for bFGF. These results imply that the agranular cell types are the main producer and/or target of rat pituitary bFGF, and that bFGF may play a role in the differentiation of pituitary endocrine cells. PMID- 8240856 TI - Early development of the pineal photoreceptors prior to the retinal differentiation in the embryonic rainbow trout, Oncorhynchus mykiss (Teleostei). AB - The development of the pineal and retinal photoreceptors in the embryonic rainbow trout, Oncorhynchus mykiss was investigated by means of light and electron microscopy. The pineal photoreceptors endowed with photoreceptive outer segments and signal-transmitting synapses appeared first 15 days after fertilization, and well developed by 21 days. In contrast, the retinal photoreceptors appeared as late as 27 days after fertilization (1 day before hatching). This temporal disparity in photoreceptor development has been compared with the photoperiodic formation of otolith increments, which is initiated during an early developmental stage lacking retinal photoreception. It has been suggested that the early development of pineal photoreceptors may be involved in light-dark recognition, establishment of the diel and circadian rhythms, and entrainment to the environmental photoperiod in the embryonic rainbow trout. PMID- 8240857 TI - Morphological profiles of mouse ovarian follicles: extensive accumulation of a strongly negative-charged substance at specific foci in follicular tissue during oocyte maturation. AB - Structural changes in cumulus-oocyte complexes and granulosa cells were induced by administering pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) to female mice. Three and 9 h after hCG administration, dissolution of the germinal vesicle (GVBD) and progression to the second metaphase occurred respectively in oocytes. The number of cumulus cells associated with the granulosa cell layer decreased significantly 1 h after hCG administration. Expansion of cumulus cell investment was due to the abundant deposition of intercellular materials in the cumulus-oocyte complexes during oocyte maturation. A strongly negative-charged (colloidal iron-positive) substance was detected in the intercellular spaces of follicular tissue, especially in the cumulus mass. Moreover, cells located where the cumulus mass and granulosa cell layer intertwined became enlarged during the resumption of the meiosis of oocytes. Colloidal iron-positive substances accumulated extensively within the intercellular spaces of the enlarged cells. PMID- 8240858 TI - Recent advances in the management of vesico-ureteric reflux. AB - Recent studies have demonstrated that both congenital hypoplasia and acquired scarring are involved in the parenchymal lesions associated with reflux nephropathy. Medical therapy can prevent symptomatic infection. While there is no proof that either medical or antireflux surgery prevents acquired scarring, paradoxically there is evidence that surgery adds no benefit to medical therapy, and that the results of medical therapy and surgical therapy are similar in children with isolated severe reflux. The group at most severe risk of renal scarring is infants and the effects of medical and surgical therapy in preventing acquired renal injury in this group have not been sufficiently investigated. On the basis of this information it has been our practice to maintain urine sterility using continuous antibiotic prophylaxis throughout infancy and early childhood. Following the development of reliable urine toilet habit and the ability to collect midstream urine specimens, antibiotics are given according to the frequency of urine infection, and weekly testing of morning urine with nitrite strips at home is used for early detection of infection and prevention of symptomatic infection. Antireflux surgery is mandatory for those children with complicated VUR (such as urinary tract obstruction) and should otherwise be reserved for those having persistent breakthrough infections in infancy and early childhood. PMID- 8240859 TI - Antibodies in the diagnosis and management of coeliac disease. PMID- 8240860 TI - Emotional and behavioural problems experienced by children living in single parent families: a pilot study. AB - This study compared the prevalence of emotional and behavioural problems experienced by 10-11 year old and 14-15 year old children living in single-parent and two-parent families in South Australia. The 10-11 year old male children living in single-parent families were found to have significantly more emotional and behavioural problems than 10-11 year old male children living in two-parent families. In particular, the younger male children living in single-parent families experienced more externalizing behaviour problems than younger male children living in two-parent families. It is suggested that paediatricians and general practitioners need to be aware of children who are at particular risk for developing emotional and behavioural problems, and the approaches which may be employed to provide help. PMID- 8240861 TI - The incidence of congenital heart defects in the first year of life. AB - Data on the incidence of congenital heart defects (CHD) in the first year of life were collected on a cohort of infants born between 1981 and 1984 in New South Wales and the Australian Capital Territory. There was a total of 1479 cases among 343,521 births, an incidence of 4.3 per 1000 livebirths. The following results were obtained after restricting the analysis to cases diagnosed definitively by echocardiography, cardiac catheterization, operation or autopsy. A significant association was found between older maternal age and having an infant with CHD, both before (RR = 1.27, CI = 1.13, 1.44) and after (RR = 1.17, CI = 1.02, 1.33) excluding chromosomally related cases. Significant associations were found between having an infant with ventricular septal defect and Italian parentage (RR = 2.50, CI = 1.11, 5.65), and for having an infant with coarctation of the aorta and Lebanese parentage (RR = 3.82, CI = 1.71, 8.52). The incidence of CHD in this Australian population is similar to overseas studies that used comparable diagnostic criteria and ascertainment. An active surveillance system for CHD is recommended as is further investigation of the factors associated with having an infant with CHD. PMID- 8240862 TI - Pneumomediastinum complicating childhood acute lymphoblastic leukaemia. AB - A case of pneumomediastinum that developed in a 10 year old girl receiving induction chemotherapy for acute lymphoblastic leukemia is reported. Three factors were identified that may have been associated with this complication: the patient suffered recurrent vomiting during her induction chemotherapy; she had travelled by air the day before the pneumo mediastinum was diagnosed; and was septic with Enterobacter at time of diagnosis. The pneumomediastinum resolved over 2 weeks without specific treatment and without further complications. PMID- 8240863 TI - A prospective study of the psychological adjustment of parents and families of children with cancer. AB - This study compared the psychological adjustment of parents and families of children with cancer, with the adjustment of parents and families in the community. In the weeks after their children's diagnoses, the mothers of children with cancer reported significantly more anxiety and insomnia, somatic symptoms and social dysfunction than mothers in the community. These problems had declined by the time of a 1 year follow-up assessment. However, at the follow-up assessment the mothers of children with cancer reported significantly more symptoms of depression and somatic symptoms than mothers in the community. A similar pattern of findings was evident among the fathers although the fathers generally reported less distress than was reported by the mothers. The results also suggest that a year after the children's diagnoses, the families in which there was a child with cancer were functioning less effectively than the families in the community. PMID- 8240864 TI - Infant onset subacute necrotizing encephalomyelopathy (Leigh's disease) AB - We reviewed six cases of proven or probable subacute necrotizing encephalomyelopathy with an onset under 12 months of age. All children had been investigated at the Adelaide Children's Hospital in the period 1975-90. Seizures (five of six) and cortical blindness (five of six) were more prominent clinical features at presentation than the literature would suggest, while respiratory abnormalities and developmental delay were also frequent. Flash visual evoked responses, brain-stem auditory evoked responses, and the interictal electroencephalogram did not contributed to diagnosis. Computerized tomography brain scans were abnormal in three of four cases with typical basal ganglia lesions in one case and brain atrophy in two cases. The diagnosis was suspected in four cases with raised blood or cerebrospinal fluid lactate concentrations. The importance of obtaining a blood or cerebrospinal fluid lactate in all infants with unexplained seizures, cortical blindness or apnoea is emphasized. PMID- 8240865 TI - Eucalyptus oil poisoning in childhood: 41 cases in south-east Queensland. AB - Forty-two cases of oral eucalyptus oil poisoning in children under 14 years of age were identified in a defined population between 1 July 1984 and 30 June 1991, and 41 were subjected to retrospective case note analysis. Thirty-three children (80%) were entirely asymptomatic. This group included all of the four children reported to have ingested more than 30 mL of eucalyptus oil. Only two of the remaining children had symptoms or clinical signs on presentation to hospital. No child required advanced life-support. There was no correlation between the amount of eucalyptus oil taken and the presence of symptoms. If the estimated volume ingested is large, or symptoms are evident, on presentation at hospital gastrointestinal decontamination should lead to a good outcome with few clinical problems. Eucalyptus oil may be a less toxic compound than has previously been believed. PMID- 8240866 TI - Infantile hypertrophic pyloric stenosis: a clinical review from a general hospital. AB - A review of 212 cases of infantile hypertrophic pyloric stenosis (IHPS) in a general hospital during an 8.5 year period documents clinical features and suggests refinement of treatment. Features such as sex, age at onset and presentation were similar to other studies. There was a preponderance of infants born in the summer. Premature infants represented 10% of the series, and the most premature (< 33 weeks gestation) presented later after birth (40 vs 25 days). Diagnosis was clinical in 65% of cases, but 4% presented before physical examination and diagnostic tests were positive. At the time of admission only 15% had significant abnormalities of serum electrolytes (chloride < 85 mmol/L). The periumbilical incision resulted in a hidden scar. Duodenal perforation (3.3%) and wound dehiscence (1.4%) are related to technical factors and can be avoided. The timing of commencement of postoperative feeds did not influence the rate of vomiting. The average length of stay was 3.5 days. These results are discussed with suggestions about how to minimize diagnostic tests and length of hospital stay. PMID- 8240867 TI - Immunization status and demographic characteristics of New Zealand infants in the first 6 months of life. AB - In the Royal New Zealand Plunket Society's 1990-91 cohort study, of 3902 children, 985 (25.2%) had fallen behind the immunization schedule by 6 months of age. These children were more likely to be from lower socio-economic groups or to have mothers who were older, with high or low education, or of higher parity. The infants were also more likely to be from non-European families, or to have unemployed fathers. Eight hundred and ten (82.2%) of the incompletely immunized children at that age could be brought up-to-date with their immunizations by a single visit to the doctor. The most common reason for delaying immunization was that the baby was sick. This was a false contraindication in 69.2, 79.0 and 78.4% of these children at the 6 week, 3 month and 5 month immunization, respectively. PMID- 8240868 TI - The epidemiology of incomplete childhood immunization: an analysis of reported immunization status in outer western Sydney. AB - We surveyed parents of children enrolled in kindergarten in 1992 in outer western Sydney and the Blue Mountains. Using parents' reports, we determined the prevalence of immunization uptake for children starting school, compared the prevalence of immunization uptake among Catholic, government and independent schools, and identified immunization providers. We also documented parental beliefs about immunization and their influence on immunization status, and identified risk factors for incomplete immunization. Nearly 89% of children were reported to be fully immunized. Immunization status did not vary significantly among the different types of school. General practitioners provided 84% of all immunizations and local councils 11%. Incomplete immunization was associated with more negative beliefs in immunization, with post-secondary education and with families who do not speak English at home. Reminder letters had little effect on immunization status. PMID- 8240869 TI - Double-barrelled tubular duplication of the ileum. AB - A three year old boy with a 10 cm double-barrelled tubular duplication of the distal ileum is reported. He presented with a small bowel obstruction after a large meal of mandarin oranges. At laparotomy an obstruction of both lumens was found which communicated at both ends with the normal ileal lumen. Although cystic duplications of the small intestine are common, tubular duplications are relatively rare, especially an obstruction with impacted undigested foods. PMID- 8240870 TI - Oesophageal duplication cyst: another cause of neonatal respiratory distress. AB - Oesophageal duplication cysts in infants (usually males) should be recognized as a cause of respiratory distress which worsens during the neonatal period, and which is associated with feeding difficulties. Their radiological appearance is that of a right-sided posterior mediastinal mass, which is shown to be cystic on ultrasound. Excision is curative. PMID- 8240871 TI - In utero repair of diaphragmatic hernia. AB - This report describes the first in utero repair of diaphragmatic hernia in Australia. The patient was a 32 year old woman with major infertility problems who was diagnosed at 15 weeks gestation as having an infant with diaphragmatic hernia. After extensive consideration and counselling the parents requested in utero repair. This was performed at 28 weeks gestation and was technically successful, but the infant was found to be dead after uterine closure. The mother has subsequently been delivered of normal twins at term by Caesarean section. In utero repair of diaphragmatic hernia requires a high degree of team work, is technically demanding and has major ethical implications. It should be restricted to nationally designated units. PMID- 8240872 TI - Erroneous measurement of arterial oxygen saturation. PMID- 8240873 TI - The decline (but not fall) of rubella. PMID- 8240874 TI - Sun overexposure and temporal cell arteritis polymyalgia. PMID- 8240875 TI - Vitamin K and childhood cancer. PMID- 8240876 TI - Personal health records. PMID- 8240877 TI - Advances in AIDS Vaccine Development: 5th Annual meeting of the National Cooperative Vaccine Development Groups for AIDS. Chantilly, Virginia, August 30 September 3, 1992. Proceedings and abstracts. PMID- 8240878 TI - The role of the nurse practitioner in the diagnosis and management of bulimia nervosa, Part 2: Physiologic management. AB - Physiologic management of bulimia nervosa is targeted primarily toward restoration and maintenance of nutrition and prevention and treatment of complications. The intensity, frequency, and duration of care depend on the severity of the disorder. Some form of physiologic management is recommended while the client is in psychotherapy. The client should be monitored for the possibility of hospitalization. Consistent management will allow for better health care. PMID- 8240879 TI - The patient with an automatic implantable cardioverter defibrillator. AB - Sudden cardiac death is the leading cause of death in the United States. A relatively new technology used to treat ventricular dysrhythmias that lead to sudden cardiac death is the automatic implantable cardioverter defibrillator. This device uses patches on the heart to deliver an energy current to convert lethal dysrhythmias. The nurse practitioner can expect to encounter these devices when seeing patients for a variety of diagnoses. This article will serve as a resource for clinical management and patient education. PMID- 8240880 TI - Blood pressure and mood responses in hypertensive patients on antihypertensive medications. AB - Nurse practitioners who are responsible for the clinical management of patients with hypertension must choose antihypertensive medications that minimize side effects. Nurse practitioners must be aware of the drugs' potential influences on physiologic and psychological variables and include adequate assessment of these responses in patients during treatment. Pindolol, propranolol, and hydrochlorothiazide were equally effective in reducing systolic and diastolic blood pressure in essential hypertensives, although pindolol and hydrochlorothiazide did not affect resting heart rate (an advantage in some patients). Hydrochlorothiazide was associated with a trend for several negative mood changes in black subjects; depression scores of black subjects were significantly increased with hydrochlorothiazide. The results of this study indicate the need to consider race and other factors in research about antihypertensive mood responses. PMID- 8240881 TI - Community health workers and primary health care in Honduras. AB - Community participation and utilization of community health workers (CHWs) are essential components of the primary health care model. The success of CHWs is dependent on their training and subsequent community support. Community-prepared nurses are ideal CHW educators. A training program for CHWs was implemented in Honduras emphasizing the principles of adult learning and problem-based learning. Following a 4-month program of training a primary health care clinic was opened and managed by CHWs for a population over 10,000. Approximately 80% of local health problems were managed by the CHWs proving that well-trained CHWs can have a significant impact on the delivery of health care. PMID- 8240882 TI - Ankle sprains. PMID- 8240883 TI - Drug Enforcement Administration registration for nurse practitioners: what it is; what it is not. PMID- 8240884 TI - Lumbar percutaneous automated nucleotomy. Technique, patient selection and preliminary results. AB - The purpose of this study was to evaluate 2 years' experience with percutaneous automated nucleotomy. Adult patients with small to medium sized disk hernias corresponding to clinical symptoms, and without evidence of free fragments or stenosis were treated on an outpatient basis. All patients had sciatica and conservative treatment had failed for at least 3 months. Using the Nucleotome R system, access to the disk was achieved in all but 6 of 172 disks in 152 patients. Degenerative disk disease or pain was the reason for technical failures. Four other procedures were discontinued because of pain and moderate hemorrhage. Except for one case of diskitis, no serious complications occurred. Of the patients in a prospective study, 63 had a follow-up period of 4 months or more. The overall clinical success rate was 62%, and was not significantly influenced by patient sex or age, duration of symptoms, level treated, disk degeneration or amount of nucleus material removed. The results are promising; however, this study indicates a need for refined patient selection in order to decrease the number of failures. PMID- 8240885 TI - Detection of breast cancer metastases in the cervical spine. AB - Thirty women with breast cancer and indications of possible cervical spine metastases underwent skeletal scintigraphy, conventional radiography, MR imaging and CT. By combining all examinations a diagnosis was settled in each patient and the different imaging techniques were compared. For all of the 26 patients with metastases in the cervical spine the correct diagnosis was found with MR imaging and CT. Both modalities also revealed paravertebral tumour growth, MR (n = 8) more than CT (n = 3). The sensitivity of skeletal scintigraphy in detecting metastases in the cervical spine was rather low, but as these examinations cover the whole body they are still convenient screening procedures, and combined with conventional radiography of selected areas are probably sufficient in many cases. Supplementary examinations with CT or MR would be valuable in patients with equivocal findings at screening for metastases or with clinical indications of metastases that remain unexplained after scintigraphy and radiography. PMID- 8240886 TI - Vertical field MR imaging of upper thorax and spine in small children. Evaluation of a new surface coil. AB - To improve image quality in a vertical field MR imaging unit, operating at low field strength (0.3 T), we have designed a half-elliptical coil for use in the upper thoracic region of small children. Our intention was also to shorten the examination time, which until now has been long, because several scans with different coils have been necessary to cover the thoracic region. The experimental coil is designed so that a child's shoulders fit into the central region. The coil consists of 2 serially connected cable-loops, mounted on a foam rubber vest. The coil performance was tested in a phantom and improvements relative to standard coils were demonstrated in in vivo studies. The results indicate that by using the half-elliptical coil, the signal-to-noise (S/N) ratio can be improved by a factor of 2 to 3 in the thoracic region of a child. PMID- 8240887 TI - MR evaluation of tuberculous spondylitis. AB - A total of 29 proven tuberculous spondylitis patients underwent MR studies. Gd DTPA enhancement was performed in 10 patients. Contiguous 2 vertebral involvement, subligamental spread of paraspinal abscesses and cord indentation were observed in 93% of the cases. Destruction of the vertebral body occurred in 76%. Intermediate or low T1 signal intensity and high T2 signal intensity were observed by MR. A combination of these characteristic findings strongly suggests the diagnosis of tuberculous spondylitis. Gd-DTPA administration did not facilitate diagnosis. MR examination should be considered as the main imaging modality for patients with suspected tuberculous spondylitis. PMID- 8240888 TI - Exercise-induced changes in magnetization transfer contrast of muscles. AB - Magnetization transfer contrast (MTC) technique provides a new type of contrast in MR imaging. The MTC method is based on the interaction between the spins of free protons and those with restricted motion. Exercise-induced changes in signal intensity and MTC were measured in the forearm muscles of 10 volunteers at 0.1 T. There was 26% increase in signal intensity of active flexor muscles after exercise when imaged with ordinary gradient echo sequence. Despite this marked intensity increase, the postexercise values of MTC did not differ from the preexercise ones. PMID- 8240889 TI - Radiology of the ileal J-pouch-anal anastomosis (IPAA). AB - Forty-four consecutive patients operated on with ileal J-pouch-anal anastomosis (IPAA) and diverting ileostomy were examined with barium contrast medium of the pouch before closure of the ileostomy. CT was performed in 4 of the patients. The anatomy of the ileal reservoir as well as complications were assessed. As normal postoperative anatomy we found a "blind loop" at the upper part of the reservoir in 29 patients and a contrast lucency at the anastomosis between the anal channel and the pouch in 5 patients. Complications were revealed at barium contrast medium examinations in 13 patients, including stenoses at the anastomosis between the pouch and the anal channel in 8 patients and fistulas in 5 patients. CT was especially valuable in the exact diagnosis and location of a pelvic abscess in 3 patients, none of which was shown at pouchography. PMID- 8240890 TI - CT for assessment of pancreatic and periampullary cancer. AB - Over a period of 2 years, 77 patients, strongly suspected of having pancreatic cancer, had abdominal CT as part of the diagnostic work-up. The CT images were reviewed by 2 radiologists who did not know the clinical course, the surgical procedure, or the final diagnosis. The positive predictive value of CT for the diagnosis of pancreatic cancer was 92% (82-97%) and the negative predictive value 69% (41-89%). The 95% confidence limits of the positive predictive value of CT in staging the extent of periampullary cancer in 52 patients varied between 21 to 79% and 59 to 100%, with the lowest values in diagnosis of liver metastases. The confidence limits for the negative predictive values in staging varied between 17 to 56% and 67 to 94%, with the highest values concerning liver metastases. We conclude that CT cannot be used as the only diagnostic procedure for confirming or excluding the diagnosis of pancreatic or periampullary cancer, and that the staging of periampullary cancer by CT alone is too inaccurate for the evaluation of resectability. PMID- 8240891 TI - Postirradiation changes in the pelvic wall. Findings on MR. AB - MR images of 45 patients who had received radiation therapy for carcinoma of the anus or recurrent carcinoma of the rectum were reviewed with regard to postirradiation changes of the pelvic wall. High signal intensity in bone marrow on T1-weighted images due to fatty replacement was almost always observed. Presacral edema occurred in 7 of 36 patients who were examined 4 to 6 weeks after the end of irradiation and was more frequent at later studies. The pelvic wall muscles showed high signal intensity on T2-weighted images compatible with edema. This finding was most frequent on studies performed more than 6 weeks after the end of irradiation. The changes subsided more than a year after radiation therapy. To avoid an erroneous diagnosis of tumor infiltration into the pelvic wall, it is important to be familiar with the normal postirradiation changes of the presacral space and the muscles. PMID- 8240892 TI - Radiologic appearance of nonpalpable intramammary lymph nodes. AB - Sixty-four well circumscribed nonpalpable lesions, mammographically evaluated as benign and cytomorphologically verified as lymph nodes, were studied concerning mammographic appearance, size and location within the breast. The great majority, 72%, were situated within the upper-outer quadrant, but lymph nodes were also diagnosed in other areas of the breast. Of the verified lymph nodes 50 (78%) had a lower density at the center than at the periphery. These were the only well circumscribed nodules with a low density center observed in a total of 3,623 nonpalpable lesions stereotaxically needled during the same period. Three of the lymph nodes were verified histopathologically. Till now, none of the other 61 lesions has turned out to be a malignant tumor during a follow-up time from 9 to 15 years. Thus a lesion presenting roentgenologically as a well circumscribed nodule with a central area of decreased density compared to the peripheral part can be confidentially diagnosed as a lymph node by the mammograms alone and no further investigation is indicated. PMID- 8240893 TI - Hematogeneous pulmonary metastasis. An experimental study using VX-2 carcinoma in rabbits. AB - The purpose of this study was to develop an experimental model of pulmonary metastases using VX-2 tumor, and to describe sequential radiologic and pathologic findings with special attention to the interstitial changes around the metastatic nodules. Through ear veins of 25 rabbits, VX-2 tumor cell suspension was injected with 0.8-mm scalp needles. Chest radiographs were taken every other day after tumor injection. The rabbits were sacrificed at scheduled times from 3 to 30 days after tumor injection. The inflated and fixed lungs were examined with visual inspection, low-kV radiography, high resolution CT (HRCT), microradiography of the sliced lung and with histopathologic studies. Hematogeneous pulmonary metastases occurred in 24 of 25 rabbits. In addition to the metastatic nodules, perinodular and peribronchovascular infiltrations were seen on low-kV radiography, HRCT, microradiography and histologic studies. PMID- 8240894 TI - Cine-MR imaging of aqueductal CSF flow in normal pressure hydrocephalus syndrome before and after CSF shunt. AB - Reproducibility of the aqueductal CSF signal intensity on a gradient echo cine-MR sequence exploiting through plane inflow enhancement was tested in 11 patients with normal or dilated ventricles. Seven patients with normal pressure hydrocephalus (NPH) syndrome were investigated with the sequence before and after CSF shunting. Two patients exhibiting central flow void within a hyperintense aqueductal CSF improved after surgery and the flow void disappeared after shunting. One patient with increased maximum and minimum aqueductal CSF signal as compared to 18 healthy controls also improved and the aqueductal CSF signal was considerably decreased after shunting. Three patients with aqueductal CSF values similar to those in the controls did not improve, notwithstanding their maximum aqueductal CSF signals decreasing slightly after shunting. No appreciable aqueductal CSF flow related enhancement consistent with non-communicating hydrocephalus was found in the last NPH patient who improved after surgery. Cine MR with inflow technique yields a reproducible evaluation of flow-related aqueductal CSF signal changes which might help in identifying shunt responsive NPH patients. These are likely to be those with hyperdynamic aqueductal CSF or aqueductal obstruction. PMID- 8240895 TI - Cranial MR imaging and cerebral 99mTc HM-PAO-SPECT in patients with subacute or chronic severe closed head injury and normal CT examinations. AB - Eighteen patients in the subacute or chronic state following severe closed head injury with normal cranial CT scans were examined by MR and 99mTc HM-PAO SPECT. Correlations were sought between these 2 imaging modalities and the clinical outcome, as defined by the Glasgow Outcome Scale (GOS) score. Both MR and SPECT revealed cerebral damage in all patients examined but structural and functional alterations did not coincide topographically in 64.9% of lesions. Nevertheless, complementary injury patterns suggesting poor recovery were found; cortical contusions and diffuse axonal injury (MR) in conjunction with cortical and thalamic hypoperfusion (SPECT) were noticed in 8 out of 12 patients with unfavorable outcome (GOS = III and IV). The synthesis of MR and SPECT information clearly enhanced the ability both to accurately assess posttraumatic brain damage and to improve patients' outcome prediction. PMID- 8240896 TI - Combined embolization and gamma knife radiosurgery for cerebral arteriovenous malformations. AB - In a study of 46 patients with cerebral arteriovenous malformations (AVMs) the value of combining embolization and gamma knife radiosurgery was assessed. In 35 patients with large grade III to V AVMs (Spetzler-Martin system) staged combined treatment was planned. In 11 patients, radiosurgery complemented embolization for a residual AVM. The number of embolization sessions ranged from 1 to 7 (median 2). Twenty-six patients needed multiple embolization sessions. In 28 patients the grade of AVMs decreased as a result of embolization. In 16 patients collateral feeding vessels developed after embolization which made delineation of the residual nidus difficult. The time lag between the last embolization and radiosurgery ranged from 1 to 24 months (median 4). Nineteen of 35 large grade III to V AVMs were possible to treat by radiosurgery following embolization. In the 46 patients complications occurred in 9 from embolization and in 2 from radiosurgery. Two patients had transient and 9 had permanent neurologic deficits. It is concluded that embolization facilitates radiosurgery for some large AVMs and therefore this combined treatment has a role in the management of AVMs. PMID- 8240897 TI - Effect of Gd-DTPA and/or magnetic field and radiofrequency exposure on sister chromatid exchange in human peripheral lymphocytes. AB - The effects of a magnetic field, radiofrequency, and gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) on the sister chromatid exchange (SCE) in human peripheral lymphocytes were investigated. Whole blood was taken from a nonsmoker and samples were exposed to magnetic force (1.5 T) alone, and to a magnetic force combined with radiofrequency waves (63.86 MHz, specific absorption rate, SAR, 0.4 W/kg). Gd-DTPA was then added to other blood samples in varying amounts and concentrations. After exposure to the conditions described above, these blood samples were cultured for 69 hours. Slides were made for an SCE evaluation. As the concentration of Gd-DTPA added to the blood increased, the SCE frequency also increased. However, the addition of Gd-DTPA at the clinical concentrations normally used did not affect the SCE frequency. PMID- 8240898 TI - Reproducibility of quantitative arteriographic assessment of atherosclerosis in the femoral artery. AB - Results from computer-analysed angiograms in the Probucol Quantitative Regression Swedish Trial (PQRST) were analysed to determine the reproducibility of the method and any drift in the analysing system. The precision index (P(mu)) for 2 angiography series, made at 10 min intervals, of the femoral artery in 276 patients was 10.5 for lumen volume and 21.9 for roughness (edge irregularity). No difference in reproducibility was found between patients with and without symptoms of peripheral atherosclerosis or when looking at the reproducibility over years. A drift of 0.67% per year in the radiographic equipment (but not in the analysis system) was found, confirmed by use of phantoms. Computer-based analysis of femoral atherosclerosis is a reliable method for follow-up trials, giving high reproducibility even if the trial spans over several years and involves different centres. The use of phantoms is essential for checking the method over time. PMID- 8240899 TI - Image intensifier based digital chest radiography. Visibility of lesions and anatomy. AB - Contrast- and edge-enhanced digital monitor images from an image intensifier system for chest radiography were compared to conventional radiograms using visual grading analysis. Eleven observers graded the visibility of rounded opacities, the carinal region, fine line structures, and also compared the overall quality of images from 20 patients. The results showed significantly better visibility on the digital monitor for the rounded opacities and the mediastinum compared to the conventional radiograms. The overall quality of the digital images was also considered better. However, the digital images showed significantly inferior visibility for the line structures in comparison with the conventional radiograms. Our study indicates that the present contrast- and edge enhanced digital images, with a spatial resolution of 1,024 x 1,024 pixels and a contrast resolution of 8 bits, are superior to conventional radiograms for the visualization of mediastinal anatomy and rounded opacities. The clinical importance of the inferior visibility of fine line structures is not clear. PMID- 8240900 TI - Assessment of tissue viability by thermography after experimentally produced frostbite of the rabbit ear. AB - Since it is difficult and time-consuming to monitor the formation of a demarcation line in frostbite by visual inspection only, thermography was used to study experimental freezing injuries of the rabbit ear. In 8 rabbits, anesthetized with ketamine hydrochloride and xylazine hydrochloride, frostbite was induced by pressing the bottom of a glass bottle 2 cm in diameter, filled with liquid nitrogen against the shaved ear skin. Thermography was performed on both ears after 2, 24, and 48 hours, and one and 3 weeks. At 2 to 48 hours the frost-bitten area was clearly warmer than the surroundings, after one week there were ill-defined diffuse cold spots in the injured area, and after 3 weeks a cold area had become clearly demarcated with a warm zone between the cold area and the surroundings. Thermography is an easy, noninvasive method for monitoring thermal changes after experimental frostbite, but its clinical value is as yet unresolved. PMID- 8240901 TI - Hyperparathyroidism--comparison of flash imaging with spin echo MR imaging. AB - MR images of the neck were prospectively studied in 19 patients with hyperparathyroidism. Fast low angle shot (FLASH) sequence was performed in addition to T1- and T2-weighted spin echo (SE) sequences. FLASH images were obtained with 320/12/20 degrees (TR/TE/flip angle) using presaturation technique. TE of 12 ms was chosen to eliminate high signal of fat tissue. In the evaluation of detectability, a combination of T1-weighted SE and FLASH images (T1WI + FLASH) was compared with a combination of T1- and T2-weighted SE images (T1WI + T2WI). MR imaging correctly depicted 20 of 30 abnormal glands on both T1WI + FLASH and T1WI + T2WI. FLASH imaging effectively eliminated high signal of fat tissue. Nineteen abnormal glands demonstrated higher signal than surrounding tissues on FLASH images, whereas 12 glands were high-intense on T2-weighted SE images. We conclude that FLASH imaging provides improved tissue contrast and anatomic delineation and, thus, may replace T2-weighted SE imaging in the neck. PMID- 8240902 TI - Position of skin puncture in translumbar aortography. PMID- 8240903 TI - Chest radiography with synchronized exposure during inspiration in ventilator treated patients. PMID- 8240904 TI - CT appearance of diffuse alveolar hemorrhage in a patient with systemic lupus erythematosus. PMID- 8240905 TI - Gray scale resolution of MR images. PMID- 8240906 TI - The Kensey catheter. PMID- 8240907 TI - Anticholinergic agents. 4. Stereocontrolled synthesis of fluorinated acetylcholine antagonists; syntheses of the two 1-cyclohexyl-1-(4-fluorophenyl)-3 piperidyl-1-propanols and their methiodides. AB - Four new putative muscarinic antagonists, (R)-(-)- and (S)-(+)-1-cyclohexyl-1-(4 fluorophenyl)-3-piperidyl-1-propanol and their methiodides, have been synthesised. Absolute configurations have been assigned on the basis of the anticipated chirality of the products of the Sharpless asymmetric epoxidation reaction. PMID- 8240908 TI - NMR and molecular dynamics study of the tripeptide L-pyroglutamyl-L histidylglycine. AB - The 1H spectrum of L-pyroglutamyl-L-histidylglycine in DMSO-d6 and 1H and 13C NMR spectra in D2O at pH 4.26 to 8.90 have been analysed. 3JHH vicinal coupling constants were used to determine rotamer populations by means of the Karplus equation. Viable molecular geometries were obtained with the aid of molecular dynamics simulations including water as solvent. In DMSO and in aqueous solution at low pH two stable conformations were identified which both have an intramolecular hydrogen bond between the histidine side chain and the C-terminal carboxylate group. PMID- 8240909 TI - Specific nitrogen-15 labelling of leucine residues in human growth hormone. AB - Biosynthetic human growth hormone (hGH) specifically 15N labelled in the leucine residues has been obtained by recombinant DNA technology, using 15N-labelled leucine and an E. coli strain that requires leucine. It is shown that, despite the possibility of minor transaminase activity, the labelling on the whole is specific, and that the two-dimensional 1H-15N correlation NMR spectra of hGH can be greatly simplified by this methodology. PMID- 8240910 TI - Molecular biology for the practitioner. PMID- 8240911 TI - Integrins. PMID- 8240912 TI - Diseases of nutrition and metabolism. PMID- 8240913 TI - The causes of irritant dermatitis. PMID- 8240914 TI - Spontaneous regression of cutaneous tumors. PMID- 8240915 TI - Evaluation and treatment of metastatic head and neck squamous cancer of cutaneous origin. PMID- 8240916 TI - Treatment of basal cell carcinoma and squamous cell carcinoma with perineural invasion. PMID- 8240917 TI - Autoimmune blistering diseases: characterization of target antigens. AB - The development of techniques for identification of antigens has had broad application and success in the field of immunodermatology. During the past 10 years, a variety of skin autoantigens, including those found within the epidermis such as in pemphigus and those defined as extracellular matrix proteins such as in EBA, have been identified as the specific targets for autoantibody binding. The intrinsic function of all the autoantigens identified to date is adhesion, either cell-to-cell or cell-to-substrate. The binding of autoantibodies to these adhesion proteins abrogates this critical function via mechanisms not yet clearly understood. Further investigations will focus not only on the characterization of additional autoantigenic targets, but also on identifying mechanisms for the origin and pathogenesis of autoantibodies in blistering skin diseases. PMID- 8240918 TI - Photoprotection in children. PMID- 8240919 TI - Oral ulcers and erosions. PMID- 8240920 TI - Management of hemangiomas. PMID- 8240921 TI - Acne vulgaris. PMID- 8240922 TI - Management and treatment with systemic glucocorticoids. PMID- 8240923 TI - Tissue distribution of bupivacaine enantiomers in sheep. AB - rac-Bupivacaine HCl was infused intravenously to constant arterial blood drug concentrations in sheep using a regimen of 4 mg/min for 15 min followed by 1 mg/min to 24 h. At 24 h, arterial blood was sampled, the animal was killed with a bolus of KCl solution, then rapidly dissected and samples were obtained from heart, brain, lung, kidney, liver, muscle, fat, gut, and rumen. Tissue:blood distribution coefficients for (+)-(R)-bupivacaine exceeded those of (-)-(S) bupivacaine (P < 0.05) for heart, brain, lung, fat, gut, and rumen by an overall mean of 43%. Blood:plasma distribution coefficients of (-)-(S)-bupivacaine exceeded those of (+)-(R)-bupivacaine by a mean of 29% and this offset the tissue:blood distribution coefficients so that the previously significant enantioselective differences disappeared. It is concluded that although enantioselectivity of bupivacaine distribution is shown by the measured tissue:blood distribution coefficients, it is not shown when tissue:plasma water distribution coefficients are calculated, suggesting that there is no intrinsic difference between the bupivacaine enantiomers in tissue affinity. Sheep given fatal intravenous bolus doses of rac-bupivacaine had significantly greater concentrations of (+)-(R)-bupivacaine than (-)-(S)-bupivacaine in brain (P = 0.028) and ventricle (P = 0.036); these could augment the greater myocardial toxicity of this enantiomer found in vitro. PMID- 8240924 TI - Rac-flurbiprofen is more ulcerogenic than its (S)-enantiomer. AB - The most common, and sometimes life-threatening, side-effects associated with the human use of the analgesic, nonsteroidal antiinflammatory drugs (NSAIDs) are gastrointestinal. These include gastritis, ulceration, and severe bleeding. The aryl propionic acid class of NSAIDs are among the most widely used of these drugs in the world, including rac-ibuprofen, rac-flurbiprofen, and rac-ketoprofen. Marketed as racemates, it was assumed that the "inactive" (R)-enantiomers, having no cyclooxygenase inhibiting effect, also had no toxic effect. In a 30-day dose response study of (S)-, (R)-, and rac-flurbiprofen given daily over a range of doses the (R)-isomer alone proved to be without apparent gastrointestinal (GI) toxicity. On the other hand the racemate proved to be 2 to 4 times as ulcerogenic in enantiomerically equivalent doses as the (S)-enantiomer. These results have significant clinical implications. PMID- 8240925 TI - Synthesis and evaluation of the antidepressant activity of the enantiomers of bupropion. AB - The synthesis of the enantiomers of bupropion, (rac)-2-tert-butylamino-3' chloropropiophenone 1 (Wellbutrin) is described. The enantiomers were compared with the racemate in both the tetrabenazine-induced sedation model and the inhibition of uptake of biogenic amine assay. No significant differences were found in their potencies to reverse tetrabenazine-induced sedation in mice or in their IC50 values as inhibitors of biogenic amine uptake into nerve endings obtained from mouse brain. PMID- 8240926 TI - Enantioselective detoxication of optical isomers of glycidyl ethers. AB - The detoxication of the enantiomers of glycidyl 4-nitrophenyl ether (GNPE), (-) (R)- and (+)-(S)-GNPE, and glycidyl 1-naphthyl ether (GNE), (-)-(R)- and (+)-(S) GNE, by rat liver glutathione transferase and epoxide hydrolase was studied. Enantioselectivity was observed with both enzymes favoring the (R)-isomers as determined by the formation of conjugate, diol, and remaining substrate measured by HPLC. Enantiomers of GNE were detoxified by cytosolic epoxide hydrolase but those of GNPE were not. Substantial nonenzymatically formed conjugates of enantiomers of GNPE were detected showing (S)-GNPE the more reactive of the pair. PMID- 8240927 TI - Stereoselective HPLC bioanalysis of atenolol enantiomers in plasma: application to a comparative human pharmacokinetic study. AB - An enantioselective HPLC bioassay has been developed relying on extraction of (R) and (S)-atenolol from alkalinized plasma or serum (pH > 12) into dichloromethane containing 5% (v/v) 1-butanol followed by an achiral derivatization of the drug with phosgene leading to (R)- and (S)-oxazolidine-2-one derivatives. Under these conditions there was quantitative conversion of the acetamido group to the corresponding nitrile. These stable derivatives were separated on a (R,R) diaminocyclohexane-dinitrobenzoyl chiral stationary phase [(R,R)-DACH-DNB] using dichloromethane/methanol 98/2 as mobile phase. Determination limits of 0.5 ng for (R)- and 0.6 ng for (S)-atenolol could be achieved using fluorimetric detection. The assay was applied to a human pharmacokinetic study which was performed in a randomized cross-over, double-blind fashion in 12 healthy volunteers, administering single oral doses of 100 mg (R,S)-, 50 mg (R)-, and 50 mg (S) atenolol. AUC0-24 and Cmax values of (R)-atenolol were slightly but significant higher than those of (S)-atenolol. The R/S ratios were 1.09 for AUC(R)/AUC(S) and 1.03 for Cmax (R)/Cmax(S) (P < 0.01) respectively after administration of the racemic drug. However, there were no difference between AUC, Cmax, and t1/2 values of each enantiomer, whether they were administered as single enantiomers or in the form of its racemic mixture. PMID- 8240928 TI - Estimation of the number of enantioselective sites of bovine serum albumin using frontal chromatography. AB - On a column with bovine serum albumin (BSA) immobilized covalently to silica, the adsorption isotherms of the enantiomers of mandelic acid, tryptophan, 2 phenylbutyric acid, and N-benzoylalanine are measured using a buffered mobile phase. Knowing the amount of BSA immobilized on the column (36 mg), the ratio of the number of enantiomer molecules needed to saturate the enantioselective retention mechanism to the number of BSA molecules is determined. The mean of the set of eight enantiomers is 0.28. These data confirm that at most one enantioselective site exists for each BSA molecule for the kind of enantiomers studied. PMID- 8240929 TI - Separation of the enantiomers of some racemic nonsteroidal aromatase inhibitors and barbiturates by capillary electrophoresis. AB - High-performance capillary electrophoresis (HPCE) and micellar electrokinetic capillary chromatography (MECC) were applied to the resolution of racemic nonsteroidal antiaromatase drugs and intermediates. Successful results were obtained in both modes using alpha-cyclodextrin (alpha-CD), beta-cyclodextrin (beta-CD), gamma-cyclodextrin (gamma-CD), or 2,6-di-O-methyl-beta-cyclodextrin (DM-beta-CD) as chiral selectors. Depending on the structure of the solute, one of the cyclodextrins was generally better suited for resolution of the racemate. The basic solutes were analyzed under HPCE conditions, whereas the nonionizable compounds such as glutethimide (Doriden) were analyzed in MECC mode. For the azole-type antiaromatase Fadrozole, both HPCE and MECC modes could be used to achieve the separation of the enantiomers. The influence of experimental factors such as pH, the presence of organic modifier, temperature, the micelle concentration, and the concentration of the chiral selector is also discussed on the basis of the results obtained with some chiral barbiturates. The possibility of analyzing the enantiomers directly in plasma samples was also demonstrated. PMID- 8240930 TI - Enantioselectivity of esterases in human brain. AB - Subcellular fractions of three human brain specimens were found to contain esterase activities which hydrolyzed racemic oxazepam 3-acetate (rac-OXA). All three human brain preparations were highly selective toward the S-enantiomer of rac-OXA. PMID- 8240931 TI - Horses and zebras. PMID- 8240932 TI - Interleukin-8: a major mediator of acute pulmonary inflammation. PMID- 8240933 TI - A role for TGF-beta in the suppression by murine bronchoalveolar cells of lung dendritic cell initiated immune responses. AB - Effective pulmonary immune responses likely require both local antigen presenting cells (APC) and regulatory suppressor cells. Bronchoalveolar cells (BAC), which consist primarily of alveolar macrophages (AM), are poor APC in most species and are often suppressive. However, dendritic cell (DC)-enriched populations from both lung interstitium and BAC have potent APC activity as measured by their capacity to stimulate both alloantigen and antigen-induced lymphoproliferative T cell responses. To determine if BAC could down-regulate pulmonary immune responses, BAC were mixed with DC-enriched loosely adherent lung interstitial cells (LAd) in a mixed leukocyte reaction (MLR). With high numbers of BAC, MLRs were consistently suppressed and suppression was partially reversed by the addition of indomethacin and catalase. Supernatants from BAC cultured with either syngeneic or allogeneic T lymphocytes in the presence of indomethacin and catalase markedly suppressed an MLR, while supernatants from BAC cultured alone were inconsistently suppressive. Antibodies to TGF-beta completely reversed the BAC-T cell supernatant-induced suppression of the MLR. However, TGF-beta antibody only partially reversed BAC-induced suppression when BAC were added directly to MLR cultures that contained indomethacin and catalase, suggesting that, in addition to TGF-beta, prostaglandins, and H2O2, BAC in culture with LAd and allogeneic T cells also produced short-lived suppressive factors and/or mediated suppression by direct cell contact. Thus, resident BAC likely utilize multiple mechanisms including TGF-beta secretion to suppress intra-alveolar immune responses initiated by lung DC. PMID- 8240934 TI - Regulation of tumor necrosis factor-alpha receptors on macrophages: differences between primary macrophages and transformed macrophage cell lines. AB - Macrophages are a pivotal cell in the production of a variety of cytokines. In addition, macrophages express receptors on their surface which allows them to act as target cells for cytokines. The regulation of both cytokine production and cytokine receptor expression in macrophages may play a key role in modulating the processes of inflammation, injury, and repair. In this report we have studied the regulation of macrophage receptors for tumor necrosis factor-alpha (TNF alpha) on rat bone marrow-derived macrophages, rat alveolar macrophages, human monocyte derived macrophages, and the human monocyte-like cell line, U937, by a variety of immunomodulatory and inflammatory agents. U937 cells and rat and human macrophages bound TNF alpha in a saturable process, with an affinity in each cell type of approximately 1 nM. Following incubation with phorbol myristate acetate for 60 min, TNF alpha binding to all cells was decreased. Interleukin-1 treatment increased TNF alpha binding to human and rat macrophages. Interferon-gamma treatment decreased receptor activity in both human and rat macrophages, but increased TNF alpha binding to U937 cells. Treatment of human and rat macrophages with endotoxin resulted in a rapid loss of TNF alpha binding, while endotoxin treatment increased receptor expression in U937 cells. In all cases, receptor regulation was the result of a change in receptor number. Finally, following intraperitoneal injection of endotoxin, TNF alpha receptor expression was down regulated on alveolar lavage cells from rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240935 TI - Pulmonary granulomatous hypersensitivity: cell-mediated responses to embolized schistosome larvae and eggs. PMID- 8240936 TI - The role of cytokines in the pathogenesis of allergic asthma. PMID- 8240937 TI - Cytokines in lung and airways fibrosis. PMID- 8240938 TI - Regulation of type I collagen mRNA translation by TGF-beta. AB - We found that TGF-beta caused a sustained increase in type I collagen production up to 48 hr after addition to human lung fibroblast cultures. Northern analysis demonstrated that although TGF-beta increased alpha 1(I) mRNA levels 4-fold at 24 hr and 3-4-fold at 48 hr after addition to cultures, there were minimal or no effects on alpha 2(I) mRNA levels at these time points. In vitro translation of RNA derived from TGF-beta-stimulated cells yielded a 2-3-fold increase in the amount of alpha 2(I) peptide when compared with the in vitro translation of RNA from unstimulated cells. Taken together, these studies showed that TGF-beta increased the translatability of the alpha 2(I) transcript. To examine whether increased translatability of the alpha 2(I) transcript resulted from structural changes in the 5' and 3' untranslated regions, primer extension and S1 nuclease protection assays were employed. These studies demonstrated no gross structural changes in the untranslated regions of the alpha 2(I) transcript induced by TGF beta-stimulation. Overall, our results suggest the presence of a TGF-beta regulatable factor which controls translation of type I collagen mRNAs. PMID- 8240939 TI - Patterns of secretion of transforming growth factor-alpha (TGF-alpha) in experimental silicosis. Acute and subacute effects of cristobalite exposure in the rat. AB - Transforming growth factor-alpha (TGF-alpha) a cytokine having potent mitogenic activity for epithelial and mesenchymal cells, may play a role in the lung remodeling of silicosis. Lung macrophages are among the major cells producing TGF alpha in a lung tissue. A pivotal event in the cascade of pathologic events leading to pulmonary silicosis is the interaction between inhaled silica and macrophages. TGF-alpha may be critical in directing the proliferation of type II pneumocytes that characterize silicosis. An inhalation model of brief exposure of pathogen-restricted male rats to 25 mg/M3 cristobalite, a highly reactive form of silicon dioxide was used to study experimental silicosis. This model is characterized by a rapid, intense, and sustained increase in macrophages, neutrophils, and lymphocytes in both alveolar and interstitial compartments of the lung. TGF-alpha was measured in an A431 cell proliferation assay made specific with the use of anti-TGF-alpha neutralizing antiserum in epithelial lining fluid (ELF) and conditioned media harvested from cultured alveolar and interstitial macrophages. Soluble TGF-alpha levels found in ELF were slightly elevated above control values during the exposure period, then increased 5-fold during the 20 weeks after the 8-day exposure period. Secretion of TGF-alpha by macrophages was elevated during exposure to cristobalite but then fell during the early post exposure period. Marked elevations in TGF-alpha secretion from both interstitial and alveolar macrophages (10- and 12-fold, respectively) occurred 8 16 weeks after cessation of exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240940 TI - Cytokine immunotherapy of non-small cell lung cancer. PMID- 8240941 TI - Expanded programme on immunization. Poliomyelitis outbreak, 1992. PMID- 8240943 TI - Diphtheria. Two recent incidents. PMID- 8240942 TI - Rift Valley fever. PMID- 8240944 TI - Maternal and child health. Maternal mortality in the Americas. PMID- 8240945 TI - Cerebrospinal meningitis. Neisseria meningitidis strains identified in Africa. PMID- 8240946 TI - Mortality trends and leading causes of death among adolescents and young adults. 1979-1988. PMID- 8240947 TI - Ureaplasma canigenitalium sp. nov., isolated from dogs. AB - Ureaplasma strains isolated from dogs (Canis familiaris) were characterized and compared with the type strains of five previously described species of the genus Ureaplasma, Ureaplasma urealyticum (isolated from humans), Ureaplasma diversum (isolated from cattle), Ureaplasma gallorale (isolated from chickens), Ureaplasma cati (isolated from cats), and Ureaplasma felinum (isolated from cats). The canine strains hydrolyzed urea but not arginine or glucose, were membrane bound, lacked a cell wall, passed through 450-nm-pore-size membrane filters, required cholesterol for growth, and formed minute colonies (diameter, 20 to 140 microns) on agar medium. These canine ureaplasma strains have been reported to be members of four serovars. The four serovars of canine strains fell into a single group on the basis of their genomic properties, as determined by DNA-DNA hybridization. On the basis of these findings, we propose that ureaplasmas with these characteristics belong to a new species, Ureaplasma canigenitalium, with strain D6P-C (= ATCC 51252) as the type strain. PMID- 8240948 TI - Classification of citrobacteria by DNA hybridization: designation of Citrobacter farmeri sp. nov., Citrobacter youngae sp. nov., Citrobacter braakii sp. nov., Citrobacter werkmanii sp. nov., Citrobacter sedlakii sp. nov., and three unnamed Citrobacter genomospecies. AB - DNA relatedness studies (hydroxyapatite method) were done on 112 strains of citrobacteria. By using the recommended definition of a genomospecies 11 genomospecies were identified in the genus Citrobacter. These genomospecies were separable by their biochemical profiles. Citrobacter koseri (Citrobacter diversus) and Citrobacter amalonaticus proved to be homogeneous species, as previously described. C. amalonaticus biogroup 1, as described by Farmer et al. (J. Clin. Microbiol. 21:46-76, 1985), was shown to be a separate homogeneous species, which was named Citrobacter farmeri sp. nov. The Citrobacter freundii complex was quite heterogeneous. C. freundii sensu stricto, as represented by the type strain, contained only 9 of 66 strains in this complex. The remaining 57 strains were members of seven genomospecies. Genomospecies 5, containing 21 strains, was named Citrobacter youngae sp. nov. Genomospecies 6, containing 15 strains, was named Citrobacter braakii sp. nov. Genomospecies 7 and 8, each containing six strains, were named Citrobacter werkmanii sp. nov. and Citrobacter sedlakii sp. nov., respectively. Genomospecies 9, 10, and 11, each containing three strains, were not named. PMID- 8240949 TI - Genetic relatedness of Bordetella species as determined by macrorestriction digests resolved by pulsed-field gel electrophoresis. AB - The genetic relatedness of the three medically important Bordetella species was examined by macrorestriction digestion of DNA with the rarely cutting enzyme XbaI and resolution by pulsed-field gel electrophoresis. Our data showed that Bordetella pertussis, Bordetella parapertussis, and Bordetella bronchiseptica produced species-specific macrorestriction profiles and that there was some variation between different isolates of the same species. Conserved bands at 130 and 155 kb occurred with B. pertussis (130 isolates tested), but the nine variable bands between 200 and 412 kb distinguished 21 types. The 10 isolates of B. parapertussis tested produced up to 11 bands at 118 to 375 kb, 4 of which were variable, giving three types. The eight B. bronchiseptica isolates examined produced up to 16 hands at 118 to 394 kb, 11 of which were variable, giving three types. The results of this work are compared with the results of previous DNA-DNA hybridization and multilocus enzyme electrophoresis studies which suggested that these three species are closely related and should be considered members of the same species. PMID- 8240950 TI - Phylogenetic relationships between some members of the genera Deleya, Halomonas, and Halovibrio. AB - The genera Halomonas and Deleya, which constitute the family Halomonadaceae, are difficult to differentiate on the basis of phenotypic and chemotaxonomic attributes. DNA-rRNA hybridization studies have indicated that some Halomonas spp. have the same level of relationship to the type species of the genus Deleya as some Deleya spp. A phylogenetic analysis of the 16S rRNA sequences of seven members of the Halomonadaceae indicated that the members of the genera Halomonas and Deleya do not form separate monophyletic subgroups, confirming the lack of any phylogenetic support for retention of these taxa as separate genera. A phylogenetic analysis of the 16S rRNA sequence of Halovibrio variabilis confirmed that this species belongs in the Halomonadaceae. All of the members of the Halomonadaceae examined and Halovibrio variabilis possess a cytosine residue at position 486 (Escherichia coli numbering), which is an extremely rare attribute among the prokaryotes and has been reported in only one other species, Listonella anguillarum. Several other signature characteristics which define this group in the gamma subclass of the Proteobacteria are identified. The Jukes-Cantor distances between members of the family Halomonadaceae, including Halovibrio variabilis, range from 0.086 to 0.000 (the levels of similarity between the 16S rRNA sequences range from 92.6 to 100%). The members of the genera Halomonas, Deleya, and Halovibrio form a monophyletic group and share common chemotaxonomic and phenotypic characteristics. Subgroups containing members of the genera Halomonas, Deleya, and Halovibrio cannot be resolved on the basis of phylogenetic, chemotaxonomic, or phenotypic data. Our data indicate that the members of the genera Halomonas, Deleya, and Halovibrio should be united in a single genus. PMID- 8240951 TI - Neisseria weaveri sp. nov. (formerly CDC group M-5), from dog bite wounds of humans. AB - The taxonomic relationships of strains belonging to Centers for Disease Control group M-5 were examined. Previous studies of rRNA cistron similarities placed this organism on the Neisseriaceae rRNA branch of rRNA superfamily III; the closest neighbors included the genus Neisseria and groups EF-4a and EF-4b. The group M-5 strains were characterized by a range of phenotypic tests, and their G + C contents and DNA-DNA relatedness levels were determined. In addition, a numerical taxonomic analysis of the whole-cell protein patterns (obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) of group M-5 and related taxa was performed. The strains studied included 45 group M-5 strains, the type strains of six Neisseria species or subspecies, three group EF-4a reference strains, and three group EF-4b reference strains plus the type strain of the phenotypically similar organism Oligella urethralis. Our results showed that the group M-5 strains were members of a homogeneous taxon distinct from phylogenetically closely related taxa. The genomic divergence as revealed by levels of rRNA cistron similarity and phenotypic characteristics indicate that group M-5 can be considered a new species of the genus Neisseria. We therefore propose the new species Neisseria weaveri, with NCTC 12742 (= CCUG 4007 = ISL775/91 = LMG 5135) as the type strain. N. weaveri strains are strictly aerobic, gram-negative, nonmotile, rod-shaped organisms which are catalase and oxidase positive, nonsaccharolytic, and able to grow on MacConkey agar and do not reduce nitrate but generally reduce nitrite.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240952 TI - Proposal for rejection of Agrobacterium tumefaciens and revised descriptions for the genus Agrobacterium and for Agrobacterium radiobacter and Agrobacterium rhizogenes. AB - The 16S rRNA sequences of seven representative Agrobacterium strains, eight representative Rhizobium strains, and the type strains of Azorhizobium caulinodans and Bradyrhizobium japonicum were determined. These strains included the type strains of Agrobacterium tumefaciens, Agrobacterium rhizogenes, Agrobacterium radiobacter, Agrobacterium vitis, Agrobacterium rubi, Rhizobium fredii, Rhizobium galegae, Rhizobium huakuii, Rhizobium leguminosarum, Rhizobium loti, Rhizobium meliloti, and Rhizobium tropici. A phylogenetic analysis showed that the 15 strains of Agrobacterium and Rhizobium species formed a compact phylogenetic cluster clearly separated from the other members of the alpha subclass of the Proteobacteria. However, Agrobacterium species and Rhizobium species are phylogenetically entwined with one another, and the two genera cannot be separated. In the Agrobacterium species, the strains of biovar 1, biovar 2, Agrobacterium rubi, and Agrobacterium vitis were clearly separated. The two biovars exhibited homogeneity in their phenotypic, chemotaxonomic, and phylogenetic characteristics, and two species should be established for the two biovars. We considered the nomenclature of the two biovars, and revised descriptions of Agrobacterium radiobacter (for the biovar 1 strains) and Agrobacterium rhizogenes (for the biovar 2 strains) are proposed. The name Agrobacterium tumefaciens is rejected because the type strain of this species was assigned to Agrobacterium radiobacter, and consequently the description of the genus Agrobacterium is revised. PMID- 8240953 TI - Amycolatopsis alba sp. nov., isolated from soil. AB - A new Amycolatopsis species isolated from soil produces a new glycopeptide antibiotic related to vancomycin. Traditional taxonomic methods and contemporary fatty acid analysis techniques were used to establish the position of this species. The hyphae fragment extensively when the organism is cultured in liquid media. The organism is characterized by white aerial hyphae that bear long chains of cylindrical conidia. The reverse side is yellowish brown; a faint light brown soluble pigment is occasionally produced. The organism has a type IV cell wall (meso-diaminopimelic acid), a type A whole-cell sugar pattern, and a type PII phospholipid pattern. Mycolic acids are not present in whole-cell hydrolysates. The major menaquinone is MK-9(H4); there is also a minor amount of MK-8(H4). The name proposed for this new species is Amycolatopsis alba. The type strain is strain A83850 (= NRRL 18532). PMID- 8240954 TI - Mycoplasma imitans sp. nov. is related to Mycoplasma gallisepticum and found in birds. AB - A mycoplasma designated strain 4229T (T = type strain) was isolated in 1984 from the turbinate of a duck in France, and similar strains were isolated from geese in France and from a partridge in England. All of these strains were originally identified as Mycoplasma gallisepticum by immunofluorescence and growth inhibition tests, but subsequent serological and molecular studies indicated only a partial relationship to this species and DNA-DNA hybridization studies revealed only approximately 40 to 46% genetic homology with M. gallisepticum PG31T. In this study morphological, cultural, and physical investigations were carried out on strain 4229T and partridge strain B2/85, after we first demonstrated the similarity between these organisms by performing a restriction enzyme analysis of their DNAs. Both strains had phenotypic properties very similar to M. gallisepticum properties, including the presence of an attachment organelle. As a result of these investigations, the organisms were assigned to the class Mollicutes, the order Mycoplasmatales, and the genus Mycoplasma. They fermented glucose, reduced triphenyl tetrazolium chloride aerobically and anaerobically, and exhibited hemadsorption and hemagglutination, but other biochemical tests were negative. Apart from a serological cross-reaction with M. gallisepticum, these organisms exhibited no significant relationship with any previously described Mycoplasma species as determined by growth inhibition or immunofluorescence tests or with a number of additional serovars and unclassified avian strains. This Mycoplasma taxon therefore appears to be a new species, for which we propose the name Mycoplasma imitans. The type strain is strain 4229 (= NCTC 11733 = ATCC 51306).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240955 TI - Phylogeny of twenty Thermus isolates constructed from 16S rRNA gene sequence data. AB - The sequences of the 16S rRNA genes of 20 Thermus isolates were determined to a high fidelity by using automated DNA sequencing and fluorescent-dye-labelled primers. The strains tested included members of the three validly named Thermus species and representatives of major taxonomic clusters defined previously for this genus. The parsimony method was used to reconstruct the phylogeny of the strains from the aligned sequences, and a bootstrap analysis revealed a number of well-supported clades. Our results are not consistent with groupings inferred from numerical taxonomy data but support the conjecture that the genus Thermus contains more species than the three currently recognized species. PMID- 8240956 TI - Genomic heterogeneity among French Rhizobium strains isolated from Phaseolus vulgaris L. AB - Levels of DNA relatedness between strains isolated from root nodules of Phaseolus vulgaris and reference strains of different Rhizobium species were determined by performing DNA-DNA hybridization experiments (S1 nuclease method). The nine strains examined were members of three genomic groups previously delineated by a restriction fragment length polymorphism analysis among strains isolated from P. vulgaris at different sites in France. In agreement with the results of the restriction fragment length polymorphism analysis, three genomic species were found. We confirmed that one of these species corresponded to Rhizobium leguminosarum since the strain examined was 100% related to the type strain of this species. The other two species were new genomic species which were less than 21% related to reference strains belonging to other Rhizobium species, including Rhizobium etli and Rhizobium tropici, and were 18% related to each other. As determined by an analysis of partial 16S ribosomal DNA sequences, each of the genomic species was found to belong to a lineage independent from the lineages of previously described Rhizobium species. Nevertheless, they were included in the group formed by the fast-growing Rhizobium species. Both genomic species 1 and genomic species 2 contained a majority of strains which were capable of nodulating both P. vulgaris and Leucaena leucocephala, like R. tropici. However, they also contained strains with a nodulation phenotype restricted to P. vulgaris, like R. leguminosarum bv. phaseoli and R. etli bv. phaseoli. Our data are the first evidence that in Europe species other than R. leguminosarum nodulate P. vulgaris. PMID- 8240957 TI - Riemerella anatipestifer gen. nov., comb. nov., the causative agent of septicemia anserum exsudativa, and its phylogenetic affiliation within the Flavobacterium Cytophaga rRNA homology group. AB - The phylogenetic position of the causative agent of septicemia anserum exsudativa, now most often referred to as [Moraxella] anatipestifer (brackets indicate a generically misnamed taxon) or "[Pasteurella] anatipestifer," was established by performing rRNA cistron similarity studies. [Moraxella] anatipestifer belongs to rRNA superfamily V, together with the genera Flavobacterium, Cytophaga, Flexibacter, Weeksella, Capnocytophaga, and Sphingobacterium. The detailed structure of rRNA superfamily V, which now contains five major rRNA homology groups, is described. An analysis of various phenotypic parameters, including new data (cellular proteins and fatty acids) and previously published data (respiratory quinones, enzyme activities, and classical phenotypic features), revealed that [Moraxella] anatipestifer differs in many aspects from its closest relatives, Flavobacterium indologenes, Flavobacterium gleum, Flavobacterium indoltheticum, Flavobacterium balustinum, Flavobacterium meningosepticum, and Weeksella zoohelcum. The combined genotypic and phenotypic data indicate that this organism should be placed in a separate genus; the name Riemerella anatipestifer gen. nov., comb. nov. is proposed for this bacterium. The specific epithet anatipestifer is kept in order to avoid nomenclatural confusion. However, it should be emphasized that the illness caused by this organism is a septicemic disease which is not restricted to ducks. PMID- 8240958 TI - Proposals to unify the genera Bartonella and Rochalimaea, with descriptions of Bartonella quintana comb. nov., Bartonella vinsonii comb. nov., Bartonella henselae comb. nov., and Bartonella elizabethae comb. nov., and to remove the family Bartonellaceae from the order Rickettsiales. AB - DNA hybridization data (hydroxyapatite method, 50 to 70 degrees C) indicate that Rickettsia prowazekii, the type species of the type genus of the family Rickettsiaceae, is substantially less closely related to Rochalimaea species than was previously thought. The levels of relatedness of Rickettsia prowazekii to Rochalimaea species and to Bartonella bacilliformis under optimal conditions for DNA reassociation were 0 to 14%, with 25.5% or greater divergence in related sequences. When stringent reassociation criteria were used, the levels of relatedness were 0 to 2%. The genera Bartonella and Rochalimaea are currently classified in different families (the Bartonellaceae and the Rickettsiaceae) in the order Rickettsiales. On the basis of DNA relatedness data, previous 16S rRNA sequence data, guanine-plus-cytosine contents, and phenotypic characteristics, neither Bartonella bacilliformis nor Rochalimaea species are closely related to other organisms currently classified in the order Rickettsiales. In fact, the closest relative of these organisms is Brucella abortus. It is therefore proposed that the family Bartonellaceae should be removed from the order Rickettsiales. Previous 16S rRNA sequence data and DNA hybridization data revealed high levels of relatedness between Bartonella bacilliformis and the four Rochalimaea species, indicating that these species are members of a single genus. It is proposed that the genus Rochalimaea should be united with the genus Bartonella in the family Bartonellaceae. The name Bartonella is retained as the genus name since it has nomenclatural priority over the name Rochalimaea. This means that new combinations for the Rochalimaea species must be created. Proposals are therefore made for the creation of Bartonella quintana comb. nov., Bartonella vinsonii comb. nov., Bartonella henselae comb. nov., and Bartonella elizabethae comb. nov. PMID- 8240959 TI - Description of two morphotypes of Peptostreptococcus micros. AB - Peptostreptococcus micros is often isolated from abscesses in several parts of the human body. The oral cavity is considered the natural habitat for the species, which has been implicated as a periodontal pathogen. In plaque samples from periodontitis patients we observed the presence of a rough morphotype of P. micros in addition to the previously recognized smooth morphotype. The rough morphotype has not been described previously. Both morphotypes are frequently isolated simultaneously from the same patient. In this paper strains of both morphotypes are described. The smooth morphotype, represented by the type strain, grew as small, dome-shaped, bright white, nonhemolytic colonies. The rough morphotype grew as equally white dry colonies which were hemolytic and had wrinkled edges. DNA-DNA reassociation studies revealed homology at the species level between the two morphotypes; in addition, no differences in physiological characteristics were observed when the organisms were tested with API-32A and API ZYM kits. The rough cells had long, thin fibrillar structures outside the cell envelope when they were stained negatively for electron microscopy. In the smooth morphotype these structures were not present. The sodium dodecyl sulfate polyacrylamide gel electrophoresis profiles of whole-cell extracts were different for the two morphotypes. In xylene-water phase partition studies, the smooth morphotype was found to be hydrophobic, whereas the rough morphotype was found to be relatively hydrophilic. The distinct morphotypes were stable on blood agar; however, the rough morphotype changed to a nonfibrillar type with a smooth colony morphology after repeated subculturing in broth. PMID- 8240960 TI - Growth characteristics of V factor-independent transformants of Haemophilus influenzae. AB - Haemophilus influenzae is a V factor-dependent species. A plasmid conferring V factor independence in Haemophilus parainfluenzae and Haemophilus ducreyi was transferred to plasmid-free H. influenzae Rd by DNA transformation. The growth characteristics of the transformants in a complex and a chemically defined medium were compared, and the ability of several exogenous pyridine nucleotides and precursors to support growth was examined. Although the transformants appeared to be V factor independent in a complex medium, in a chemically defined medium they exhibited both V factor-dependent and nicotinamide-dependent growth. Because of the inability of the plasmid-free H. influenzae Rd to utilize nicotinamide for growth, it was concluded that the genes conferring this function were plasmid linked. Our results indicate that the V factor requirement, as it is presently defined, is not suitable to serve as a definitive taxonomic criterion for species determination in the family Pasteurellaceae. PMID- 8240961 TI - Micrococci demonstrate a phosphatase activity which is repressed by phosphates and which can be differentiated from that of staphylococci. AB - The phosphatase activities of 114 micrococcal strains belonging to seven different species and of an additional 150 unspeciated micrococcal strains were evaluated on solid media at various pHs containing or not containing phosphates. In the presence of phosphates, only nine strains (five unspeciated strains, one Micrococcus luteus strain, and three Micrococcus varians strains) yielded a positive reaction on plates at pH 8. In media (at pH 8) deprived of phosphates, in contrast, all but 15 strains demonstrated clear-cut phosphatase activity. Acid phosphatase could not be evaluated on solid media since none of the strains grew satisfactorily on plates at pH 5. The phosphatase activities of seven (one or two for each species, which included phosphatase-negative strains) of the strains whose colonies proved phosphatase negative at pH 8 and of 18 (two or three strains per species) of those with phosphatase-positive colonies were evaluated at pH 5 and 8.5 in toluene-treated cells which had been grown in liquid media at pH 7 containing or not containing phosphates. All strains demonstrated distinct phosphatase activity at both pHs when grown in media not containing phosphates. In contrast, when strains were grown in the presence of such substances, virtually no activity was observed at pH 8.5, and, generally, a much reduced activity was observed at pH 5. The phosphatase activity of micrococci of the various species (three to eight strains per species) was also compared with that of staphylococci of different species (5 to 10 strains per species) by the methyl green-phenolphthalein diphosphate method, the sensitivity of which can be varied by using different enzyme substrates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8240962 TI - Molecular systematics of the genus Zoogloea and emendation of the genus. AB - Phylogenetic relationships among strains of Zoogloea and related taxa were determined by 16S rDNA sequencing and genomic DNA hybridization techniques. The 16S rRNA gene was amplified by the polymerase chain reaction with a pair of eubacterial consensus primers and sequenced directly by using an automated fluorescent DNA sequencer. Sequence comparisons and distance matrix tree analysis revealed that Zoogloea ramigera IAM 12136 (= N. C. Dondero 106, type strain) and Zoogloea sp. ATCC 19324 formed a lineage with Rhodocyclus purpureus in the beta subclass of Proteobacteria. Z. ramigera IAM 12670 (= P. R. Dugan 115) was shown to belong to another cluster with Alcaligenes eutrophus and Pseudomonas cepacia in the beta subclass. In contrast, Z. ramigera IAM 12669 (= K. Crabtree I-16-M) proved to be a member of the alpha subclass of the Proteobacteria, closely related to Agrobacterium tumefaciens. Genomic DNA hybridization studies also showed that there is genetic diversity among the strains currently designated Z. ramigera, but typical Zoogloea strains, characterized by their production of rhodoquinones, are highly related to each other and can be regarded as a single species. On the basis of the molecular data, together with the early phenotypic and chemotaxonomic information, we have emended the generic description of Zoogloea. PMID- 8240963 TI - Phylogenetic relationship of Gemella morbillorum to Gemella haemolysans. AB - Partial 16S rRNA gene sequences (16S rDNA) of Gemella morbillorum and Gemella haemolysans were determined by sequencing polymerase chain reaction-amplified DNA. A phylogenetic analysis of 53 eubacterial 16S rRNA sequences grouped the gemellae on a distinct branch separate from the 18 members of the genus Streptococcus. DNA-DNA hybridization results indicate that the two gemellae are related at the genus level but are not a single species. PMID- 8240964 TI - Rickettsia massiliae sp. nov., a new spotted fever group Rickettsia. AB - We propose the name Rickettsia massiliae sp. nov. (type strain, Mtu1 in the Collection of the World Health Organization Collaborative Center for Rickettsial Reference, Marseille, France) for a spotted fever group rickettsia determined to be distinct from previously recognized species by the serotyping method (L. Beati, J.-P. Finidori, B. Gilot, and D. Raoult, J. Clin. Microbiol. 30:1922-1930, 1992). This rickettsia has biological characteristics similar to those of the other spotted fever group rickettsiae. In addition, a sodium dodecyl sulfate polyacrylamide gel electrophoresis protein analysis, a polymerase chain reaction followed by a restriction fragment length polymorphism analysis of DNA fragments, and pulsed-field electrophoresis of the genome of R. massiliae revealed unique migration patterns distinct from those of all previously recognized spotted fever group rickettsiae. These additional characteristics (Beati et al., J. Clin. Microbiol. 30:1922-1930, 1992), together with the data usually considered sufficient for description of rickettsiae, are crucial to the proposal of this new species and should be helpful in species identification. PMID- 8240965 TI - Mycobacterium genavense sp. nov. AB - Strains of a suggested novel type of mycobacterium have been repeatedly isolated from patients with AIDS. We summarize the results of tests performed to determine enzymatic activities and metabolic properties, the results of fatty acid analyses, and the results of a comparative 16S rRNA sequence determination. We propose formally that this organism represents a new species, Mycobacterium genavense. The type strain is strain 2289, a culture of which has been deposited in the American Type Culture Collection as strain ATCC 51234. PMID- 8240966 TI - Characteristics of DNA and multiple rpoD homologs of Microcystis (Synechocystis) strains. AB - The base compositions of DNAs from nine Microcystis strains, as determined by high-performance liquid chromatography, were 41 to 42 mol% G+C. Chromosomal DNAs derived from these strains were found to be extremely resistant to many restriction endonucleases, and a restriction analysis revealed the presence of a dam-like methylase or both dam- and dcm-like methylases in all of the strains examined. Genomic Southern hybridization in which a synthetic oligonucleotide probe (rpoD probe) was used showed that members of the genus Microcystis might have multiple rpoD homologs, and the hybridization signal patterns observed with the DNAs of Microcystis aeruginosa strains were different from each other. PMID- 8240967 TI - Pursuit of the Corynebacterium striatum type strain. AB - The description of Corynebacterium striatum (Chester 1901) Eberson 1918AL in Bergey's Manual of Systematic Bacteriology has many inconsistencies with the identification scheme from the Centers for Disease Control and Prevention. We have studied the four C. striatum reference strains available from the American Type Culture Collection and the National Collection of Type Cultures and found that they fit the Centers for Disease Control and Prevention description of this species. However, it appears that the wrong strains were deposited for this species, because none of the reference strains fits the descriptions in the original publications. This is a substantial case for declaring it a nomen dubium, but the name could be rescued if a careful search reveals a strain that was used in making the original description. It is hoped that some readers may have the missing strains labeled with the early names Bacillus striatus, Bacillus flavidus, or Corynebacterium flavidum. PMID- 8240968 TI - Aeromonas enteropelogenes and Aeromonas ichthiosmia are identical to Aeromonas trota and Aeromonas veronii, respectively, as revealed by small-subunit rRNA sequence analysis. AB - The 16S rRNA gene sequences of the type strains of Aeromonas enteropelogenes and Aeromonas ichthiosmia were determined by polymerase chain reaction direct sequencing in order to clarify their interrelationships with other aeromonad species. On the basis of 16S rRNA gene sequence analysis, A. enteropelogenes and A. ichthiosmia were found to be identical to Aeromonas trota and Aeromonas veronii, respectively. PMID- 8240969 TI - Cloning and characterization of alpha PS1, a novel Drosophila melanogaster integrin. AB - The Drosophila position-specific integrins (PS integrins or PS antigens) comprise two heterodimeric complexes, alpha PS1 beta PS and alpha PS2 beta PS. With the cloning of alpha PS1 described here, we complete the characterization of the primary structure of the three PS integrin subunits. We have purified the alpha PS1 subunit, obtained peptide sequence and isolated genomic and cDNA clones. The encoded alpha PS1 protein contains the cysteine pattern of the cleaved alpha integrins, three putative metal binding domains and shows the other characteristic features of alpha integrins. Regions of sequence variation indicate that alpha PS1 is distinct from all other alpha chains. The transcript analysis shows that the patterns of both alpha PS1 mRNA and protein expression are the same, suggesting that the gene is controlled transcriptionally. We compare the gene structures of the Drosophila alpha PS1, alpha PS2, the human alpha IIb and alpha X (p150,95) and the C. elegans F54G8.3 integrins. We find several positions and phases of introns conserved which, supported by conservation also in the amino acid sequence, indicates that they all derive from a common ancestral gene. PMID- 8240970 TI - Regulation of runt transcription by Drosophila segmentation genes. AB - The runt gene plays an important role in the genetic hierarchy that generates the segmented body pattern during the early stages of Drosophila embryogenesis. We studied mRNA expression in mutant embryos in order to investigate the regulation of runt transcription during these stages. We used sensitive whole-mount in situ hybridization procedures to identify the earliest, and therefore most likely direct regulatory effects. There are several distinct phases of runt expression in the early embryo. We find that each phase depends on a different set of regulators. The first phase of expression is a broad-field of mRNA accumulation in the central regions of syncytial blastoderm stage embryos. This pattern is due to terminal repression by the anterior and terminal maternal systems. The effect of the terminal system, even at this early stage, is mediated by two zygotic gap genes, tailless and huckebein. A 7 stripe pattern of runt mRNA accumulation emerges during the process of cellularization. The initial formation of this pattern depends on position-specific repression by zygotic gap genes. Examination of the early RNA patterns of the pair-rule genes even-skipped, hairy, and fushi tarazu indicate that they are also regulated in a similar manner. Three pair-rule genes, hairy, even-skipped, and runt itself, also affect runt's 7 stripe pattern. The effects of runt are stripe specific; the effects of hairy are more uniform; and the patterns obtained in even-skipped mutant embryos show a combination of both stripe specific and uniform regulatory effects. A third distinct phase of expression occurs at the onset of gastrulation when runt becomes expressed in 14 stripes. fushi tarazu plays a negative regulatory role in generating this pattern, whereas the pair-rule genes paired and odd-paired are required for activating or maintaining runt expression during these stages. PMID- 8240971 TI - Analysis of gastrulation: different types of gastrulation movement are induced by different mesoderm-inducing factors in Xenopus laevis. AB - In this paper we analyze the control of gastrulation in Xenopus laevis. Our approach takes advantage of the observation that mesoderm-inducing factors such as activin, FGF and BMP-4 induce presumptive ectodermal cells to undergo gastrulation-like movements. Activin, for example, makes intact animal pole regions undergo convergent extension and causes individual cells to spread and migrate on a fibronectin-coated substrate. By varying the concentrations of the growth factors to which animal pole cells are exposed, and by applying them in different combinations, we show how graded distributions of a combination of factors could establish the correct spatial and temporal patterns of gastrulation in the Xenopus embryo. The distributions we propose support and develop the model previously suggested by Green et al. (1992) to account for the spatial patterns of gene activation in the early embryo. PMID- 8240972 TI - A new approach of urodele amphibian limb regeneration: study of myosin isoforms and their control by thyroid hormone. AB - In P. waltlii, an urodele amphibian species which undergoes spontaneous metamorphosis, study of native myosin in pyrophosphate gels at various stages of normal development demonstrates a complete larval to fast myosin isoforms transition, which occurs more precociously in forelimb muscles than in the dorsal and ventral muscles. In the neotenic species A. mexicanum, forelimb muscles development also presents a complete myosin isoforms transition which is in contrast with the partial myosin isoforms transition observed in the dorsal muscle. In metamorphosed or neotenic animals of both species aged 1 year, forelimb regeneration is characterized by a complete transition from larval to fast myosin isoforms, that occurs earlier and more rapidly than in normal forelimb development. When forelimb regeneration is studied in P. waltlii aged 4 years, the adult fast and slow isomyosins are expressed very early in the regeneration process. In experimental hypothyroidian P. waltlii, the larval to fast isoforms transition in regenerating forelimb muscles is slightly delayed. Experimental hyperthyroidism accelerates the disappearance of larval isomyosins in regenerating forelimb muscles, both in P. waltlii and A. mexicanum aged 1 year. This work demonstrates that changes in myosin isoform pattern during forelimb regeneration in adult urodele amphibians are different from changes occurring in the normal forelimb development. They take place without any thyroid hormone influence, as opposed to normal development, and appear to be age dependent. PMID- 8240973 TI - Mutations of androgen receptor gene in androgen insensitivity syndromes. AB - The androgen receptor belongs to the family of steroid-thyroid hormone-retinoid nuclear receptors. It contains 3 major domains: a hormone-binding region, a DNA binding region and an amino-terminal region. Cloning of the cDNA encoding the androgen receptor and elucidation of the androgen receptor gene structure enabled the characterization of the molecular defects associated with androgen insensitivity. Mutations of the androgen receptor in 46,XY individuals cause a spectrum of androgen insensitivity syndromes, ranging from female phenotype (testicular feminization) to minor degrees of undervirilization or infertility. Reports on androgen receptor gene structure in patients with complete or partial forms of androgen insensitivity demonstrate that gene deletions are very rare. Several categories of mutations have been reported and are reviewed in this paper. Nucleotide substitutions in the androgen-binding domain or the N-terminal region that cause insertion of premature termination codons result in failure to form a functional protein. Missense mutations within the androgen-binding domain are responsible for a decrease or absence of receptor-binding activity. Mutations within the DNA-binding domain are associated with a positive receptor-binding form of androgen insensitivity. Analysis of described mutations indicates that they are spread throughout the gene, either associated with partial or complete androgen insensitivity. Furthermore, the same point mutation was reported to be associated with variable phenotypic expression of androgen insensitivity syndrome. It is thus difficult to define a genotype/phenotype relationship. However, mutations causing androgen insensitivity will certainly yield important new insights into the molecular basis of androgen action. PMID- 8240974 TI - Sequencing of an RNA transcript of the human estrogen receptor gene: evidence for a new transcriptional event. AB - Two transcripts of the human estrogen receptor (ER) gene have been described, ER mRNA 1 and mRNA 2, different in their 5' untranslated region. By performing reverse transcriptase-polymerase chain reaction with oligonucleotides specific for the 5' genomic region of the human ER gene we have identified a new ER RNA transcript. The sequence analysis of cDNA from MCF7 breast cancer cells and endometrial human tissues demonstrates that this transcript originates further upstream of the initiation transcription sites so far proposed. Primer extension analysis on RNA from MCF7 cells reveals in the upstream region a possible transcription start site at -3090. In agreement with this result, Northern blot analysis shows, in addition to the canonical 6.3 kb ER mRNA, an ER RNA transcript of approx. 7.4 kb in size. The presence of the additional ER mRNA suggests the existence of a new upstream 5' promoter directing transcription of the human ER gene. PMID- 8240975 TI - Tissue specific effects of thyroid hormone on 11 beta-hydroxysteroid dehydrogenase gene expression. AB - 11 beta-Hydroxysteroid dehydrogenase (11 beta-HSD) by converting active glucocorticoid to an inactive metabolite confers specificity upon the mineralocorticoid receptor (MR) and regulates ligand access to the glucocorticoid receptor (GR). Factors which influence 11 beta-HSD activity seem likely to be of considerable importance in the modulation of both mineralocorticoid and glucocorticoid hormone action. The administration of tri-iodothyronine (T3) to rats has previously been shown to reduce 11 beta-HSD activity in liver but not in kidney. We have studied the effect of T3 on 11 beta-HSD gene expression in vivo in rat liver, kidney, distal colon and pituitary. In addition the effects of T3 on 11 beta-HSD gene expression in vitro in the rat pituitary GH3 cell line have been studied. T3 administration to normal adult rats (40 micrograms/day, s.c. for 1, 3 and 7 days) resulted in a marked decline in liver and pituitary 11 beta-HSD mRNA levels and activity following 3 and 7 days of treatment. These reduced levels were maintained for 3 days following withdrawal of T3 treatment, but returned to control levels after 7 days. In contrast 11 beta-HSD mRNA and activity in kidney and distal colon were unaffected by T3 treatment at each time point studied. In vitro, levels of 11 beta-HSD mRNA and activity in GH3 cells were unchanged following 8, 24 and 72 h treatment with T3 (10(-8) to 10(-6) M). T3 bio-activity was confirmed by a marked dose-dependent decline in the expression of the T3 and glucocorticoid responsive gene, prolactin. T3 inhibits 11 beta-HSD gene expression in both liver and pituitary at a pre-translational level. This effect is absent in the predominantly mineralocorticoid target tissues, kidney and distal colon, i.e. it is tissue specific and as such is consistent with the existence of multiple differentially regulated isoforms of 11 beta-HSD. The time course of the T3 effect in liver and pituitary in vivo and the lack of any effect in vitro suggests that this action is indirect, and not as a result of interaction between the T3 receptor and the putative thyroid hormone response element on the rat 11 beta-HSD gene. PMID- 8240976 TI - Hormonal effects of turosteride, a 5 alpha-reductase inhibitor, in the rat. AB - Turosteride [FCE 26073; 1-(4-methyl-3-oxo-4-aza-5 alpha-androstane-17 beta carbonyl)-1,3- diisopropylurea] is a novel inhibitor of 5 alpha-reductase, the enzyme responsible for the conversion of testosterone (T) to 5 alpha dihydrotestosterone (DHT). The compound caused inhibition of rat and human prostatic enzymes, with IC50 values of 55 and 53 nM, respectively. In addition, turosteride did not show any relevant binding affinity to the rat prostate androgen receptor (IC50 84 microM; relative binding affinity 0.004% of DHT). The endocrine effects of turosteride were evaluated in adult male rats, treated orally at daily doses of 3, 10 and 30 mg/kg for 20 days. At these doses, the compound reduced the ventral prostate weight by 10, 33 and 42% and the intraprostatic total DHT content by 61, 74 and 78%, respectively, whereas no change in the intraprostatic content of T was observed. Turosteride caused a 40% reduction of serum DHT levels which, however, did not reach statistical significance, whereas serum T levels were similar to control animals. No effect on serum luteinizing hormone or prolactin was observed. These results indicate that the antiprostatic effect of turosteride in the adult rat is related to inhibition of the conversion of T to DHT. However, at variance with other 5 alpha reductase inhibitors (e.g. finasteride), turosteride caused a decrease in prostatic DHT not associated with a secondary increase in T content. This peculiarity of turosteride may represent an improvement of the compound over other inhibitors. PMID- 8240977 TI - Evidence that biphasic effects of basic fibroblast growth factor on 5-ene-3 beta hydroxysteroid dehydrogenase-isomerase activity in cultured immature Leydig cells are mediated by binding to heparan sulfate proteoglycans. AB - The present studies examined the effects of heparin, heparinase, insulin or insulin-like growth factor-I on basic fibroblast growth factor actions on 5-ene-3 beta-hydroxysteroid dehydrogenase-isomerase activity of cultured immature rat Leydig cells. Treatment with basic fibroblast growth factor alone (0.025-20 ng/ml) for 2 days had a biphasic effect on enzyme activity, with lower concentrations (0.025-1 ng/ml) progressively inhibiting activity to approximately 20% of control, while higher concentrations (2.5-20 ng/ml) partially reversed the inhibitive effects. The inclusion of 10 micrograms/ml heparin, a concentration reported to inhibit growth factor binding to heparan sulfate proteoglycans, blocked the increase in enzyme activity elicited by higher growth factor concentrations, but had no effect on the progressive decline in activity due to lower concentrations. Concomitant treatment with heparinase I and III, which specifically hydrolyze heparan sulfate proteoglycans, had a similar effect. In addition, both insulin and insulin-like growth factor-I partially reversed the inhibition of enzyme activity due to treatment with 1 ng/ml basic fibroblast growth factor. These studies suggest that some basic fibroblast growth factor actions on cultured immature Leydig cells are mediated by binding to heparan sulfate proteoglycans, and that both insulin and insulin-like growth factor-I can reverse the inhibitive effects on 5-ene-3 beta-hydroxysteroid dehydrogenase isomerase activity. PMID- 8240978 TI - Cytochrome P450c17 from porcine and bovine adrenal catalyses the formation of 5,16-androstadien-3 beta-ol from pregnenolone in the presence of cytochrome b5. AB - The synthesis of 5,16-androstadien-3 beta-ol from pregnenolone occurs via a cytochrome P450-dependent reaction (andien-beta synthase) that is analogous to the C17-hydroxylase/lyase reaction. It is not known whether the andien-beta synthase activity in adult porcine testis involves cytochrome P450c17 or is unique to porcine testis. Andien-beta synthase activity in testis microsomes was inhibited by high pH and concentration of salt, while C17-hydroxylase/lyase activity was stimulated under these conditions. Cytochrome P450c17 purified from adult porcine testis and adrenal glands and bovine adrenal glands had only C17 hydroxylase/lyase activity in the absence of cytochrome b5. However, when cytochrome b5 isolated from porcine testis was added, andien-beta synthase activity was detected in all three preparations of cytochrome P450c17, with the highest activity found in the porcine preparations. The andien-beta synthase activity was further increased from 2.5 to 6 times when NADH cytochrome b5 reductase was added along with cytochrome b5. Levels of mRNA for cytochrome b5 relative to cytochrome P450c17 mRNA were five times higher in porcine testis than in porcine adrenal. It appears that the andien-beta synthase activity is catalysed by cytochrome P450c17, which is not unique to the porcine testis and is dependent upon adequate levels of cytochrome b5. PMID- 8240979 TI - Steroid hormones regulate cAMP and cGMP production by porcine granulosa cells in vitro. AB - The influence of steroid hormones on the output of the cyclic nucleotides adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) by porcine granulosa cells was investigated. Both progesterone (100, 1000, 10,000 and 100,000 pg/ml medium) and estradiol (100, 1000, 10,000 and 100,000 pg/ml medium) activated cAMP and cGMP production. Testosterone (100 or 1000 pg/ml medium) also stimulated cAMP output. The stimulating effect of steroid hormones on cyclic nucleotide production may suggest the involvement of cAMP- and cGMP-dependent intracellular mechanisms in the action of steroid hormones on porcine ovarian cells. PMID- 8240980 TI - Comparative effects of short term treatment with norethisterone and sex steroids on gonadotropin secretion in rat pituitary cell cultures. AB - The short term effects of norethisterone (NET), progesterone (P), estradiol (E2) and dihydrotestosterone (DHT) on the gonadotropin secretion of pituitary cells, from both male and female rats, in primary culture primed with E2 were studied. In female cells, NET only increased the GnRH-induced secretion of LH, while P increased both LH and FSH. Male pituitary cells showed an increased response to GnRH after P pretreatment only if the E2 concentration was augmented. However with the same E2 conditions pretreatment with NET decreased the stimulated LH, but not FSH secretion. Pretreatment with E2 inhibited LH stimulated secretion from pituitary cells of male but not female rats. Furthermore DHT treatment diminished the GnRH response for both LH and FSH in pituitary cells from both sexes. Androgen pretreatment increased basal gonadotropin secretion in male but not in female cells. Basal FSH secretion was increased by NET pretreatment in male cells. This suggests that NET is metabolized by cultured pituitary cells to A-ring reduced compounds during the 4 h incubation period. The formation of NET metabolites, particularly the 3 beta, 5 alpha and 5 alpha-NET might be responsible for the estrogenic and androgenic effects observed when NET was administered to the cultured pituitary cells. PMID- 8240981 TI - Metabolism of dehydroepiandrosterone by cultured human adipose stromal cells: identification of 7 alpha-hydroxydehydroepiandrosterone as a major metabolite using high performance liquid chromatography and mass spectrometry. AB - Studies of the metabolism of dehydroepiandrosterone (DHA) by cultured human adipose stromal cells revealed that the most abundant metabolite detected by HPLC was a polar compound accounting for up to 45% of total radioactivity. This metabolite was isolated by chromatography on Lipidex 5000 from the culture medium of breast adipose stromal cells cultured with unlabelled DHA (5 microM) and identified by combined capillary gas chromatography and mass spectrometry as 7 alpha-hydroxydehydroepiandrosterone (7 alpha-OHDHA). In breast adipose stromal cells, the conversion of DHA to 7 alpha-OHDHA was linear from a substrate concentration of 10 nM to 1 microM. At 1 microM substrate concentration, the formation of 7 alpha-OHDHA in four patients ranged from 6.1 to 22.5 ng/10(5) cells/24 h. Incubations carried out in primary culture and up to the fifth subculture revealed continued formation of 7 alpha-OHDHA. Adipose stromal cells from abdomen, flank and perinephric fat also produced 7 alpha-OHDHA from DHA. These studies have shown that 7 alpha-OHDHA is a major metabolite of DHA in human adipose stromal cells. The variability from patient to patient and the magnitude of this conversion suggests that this pathway may play an important role in the peripheral metabolism of DHA. PMID- 8240982 TI - Estrogen and androgen receptor mediated stimulation and inhibition of proliferation by androst-5-ene-3 beta,17 beta-diol in human mammary cancer cells. AB - Androst-5-ene-3 beta,17 beta-diol (ADIOL) and 5 alpha-androstane-3 beta,17 beta diol (5 alpha A), which are metabolites of dehydroepiandrosterone and dihydrotestosterone, are known to have estrogenic properties. This study reevaluates the estrogenic effects of ADIOL and 5 alpha A in MCF-7 cells and demonstrates additionally androgen-like inhibitory properties of these compounds in human hormone-dependent mammary cancer cells. ADIOL and 5 alpha A (10-100 nM) stimulate the proliferation of estrogen-sensitive MCF-7 cells. Binding assays with the estrogen receptor and inhibition of stimulation with the antiestrogen tamoxifen support the involvement of the estrogen receptor. On the other hand, the mammary cancer cell line MFM-223 is strongly inhibited by ADIOL and 5 alpha A in the same concentration range. This cell line is androgen receptor positive and is inhibited by androgens, but unresponsive to estrogens and progestins. The inhibitory effects of ADIOL and 5 alpha A in MFM-223 cells are mediated by the androgen receptor as demonstrated by receptor studies and competition experiments with hormone antagonists. ADIOL and 5 alpha A thus possess estrogen- and androgen like properties and can stimulate or inhibit proliferation of human mammary cancer cells. The reactions of mammary cancer cells to these steroids depend on the receptor content and the growth properties of the individual cell line. PMID- 8240983 TI - The anti-estrogen tamoxifen blocks the stimulatory effects of interleukin-6 on 17 beta-hydroxysteroid dehydrogenase activity in MCF-7 cells. AB - Previous studies have revealed that human breast fibroblasts secrete the cytokine, interleukin-6 (IL-6) which stimulates the ability of MCF-7 human breast carcinoma cells to convert estrone (E1) to the biologically more active 17 beta estradiol (E2). This is mediated by an increase in reductive 17 beta hydroxysteroid dehydrogenase (17-HSD) activity. In the studies described here, we have extended our observations using the anti-estrogen, tamoxifen, to demonstrate that in a steady state, endogenous intracellular concentrations of E2 have no effects on reductive 17-HSD activity (E1-->E2), but are already maximally inhibitory for the oxidative reaction (E2-->E1). Increasing intracellular concentrations of E2, however, stimulated the reductive 17-HSD in a dose dependent manner. IL-6 stimulated the reductive pathway and was synergistic with E2. IL-6 is most likely acting through an E2-dependent mechanism, since tamoxifen completely reversed the effects of E2 and IL-6 separately and in combination. These observations suggest that tamoxifen may reduce intratissular levels of E2 by directly increasing oxidative 17-HSD activity and by blocking the actions of paracrine factors such as IL-6 which increase reductive 17-HSD activity. PMID- 8240984 TI - Synthesis, receptor binding and biodistribution of the gem-21-chloro-21 iodovinylestradiol derivatives. AB - Radioiodinated 11 beta-methoxy-(17 alpha,20E)iodovinylestradiol (11 beta-OMe IVE2) shows high estrogen receptor (ER)-mediated uterus uptake and good potential as an ER-imaging agent. In order to examine the tolerance of the ER for modification about the iodovinyl substituent, we prepared the (17 alpha,20Z chloro)21-chloro-21-iodovinylestradiol (4a) and several derivatives featuring 11 beta-methoxy (4b), 11 beta-ethoxy (4c) or 7 alpha-methyl (4d) substituents. All gem-dihalogen derivatives 4a-d were prepared from the 17 alpha-chloroethynyl precursors. The intermediate chlorostannylvinyl derivatives were obtained using tri-n-butyltin hydride and palladium acetate catalyst. Compounds 4a and 4b were labeled with 125I via their corresponding tin intermediates and their tissue distribution was studied in immature female rats. Addition of a 21-Cl to the 17 alpha-ethynylestradiols reduced ER binding affinity, except for the 11 beta substituted analogs which showed a pronounced increase. Surprisingly, addition of a 21-Cl to the (17 alpha,20E)IVE2 resulted in increased ER binding affinities and augmented ER-mediated uterus uptake, which may result from the pronounced increase in the dipole moment of the molecule. Thus, further modifications at the C-21 position of IVE2 are well tolerated by the ER. However, addition of the 21 Cl also resulted in increased radioiodine uptake by the thyroid, much slower blood clearance and lower uterus to blood/nontarget ratios, suggesting increased in vivo instability of the C--I bond of the gem-chlorine-iodine atoms which may reflect the increase in steric and electronic interference. PMID- 8240985 TI - Regulation of 17 beta-hydroxysteroid dehydrogenase in a newly-established human breast carcinoma cell line. AB - UISO-BCA-1 human breast carcinoma cell lines, established and characterized in our own laboratory, were used to study both oxidative and reductive pathways of 17 beta-hydroxysteroid dehydrogenase (17 beta-OH-SDH). This enzyme has been suggested to catalyze conversion of both estrone to estradiol and estradiol to estrone. In order to determine the natural preferred enzymic pathway, the enzymic activity was assayed in intact cell monolayers. In these cells, reduction of estrone to estradiol was 7-fold higher than oxidation of estradiol to estrone. For the reductive pathway, the apparent Michaelis-Menten (Km) was 5.5 microM, and for the oxidative pathway, it was 14.3 microM. The enzymic conversion of estrone to estradiol was enhanced by 72 h treatment with estrone, estradiol and R5020, dehydroepiandrosterone, or dehydroepiandrosterone sulfate. On the other hand, oxidation of estradiol to estrone was stimulated by estradiol+R5020, but inhibited by estrone treatment. The results of the kinetic study, and regulation by various steroids in the present study, indicate that oxidation of estradiol or reduction of estrone is probably mediated via different forms of 17 beta-OH-SDH. PMID- 8240986 TI - Enzyme-catalyzed synthesis of radiolabeled 5-pregnen-3 beta-ol-20-one sulfate. AB - The enzymatic synthesis of [3H]5-pregnen-3 beta-ol-20-one sulfate using [3H]5 pregnen-3 beta-ol-20-one, 3'-phosphoadenosine-5'-phosphosulfate and hydroxysteroid sulfotransferase 1 purified from rat liver is reported. The described procedure allowed the obtainment of high specific activity [3H]5 pregnen-3 beta-ol-20-one sulfate in yields ranging from 78 to 86% with respect to [3H]5-pregnen-3 beta-ol-20-one. Two-dimensional thin-layer chromatography was used to purify [3H]5-pregnen-3 beta-ol-20-one sulfate which upon solvolysis resulted in the formation of [3H]5-pregnen-3 beta-ol-20-one. The identity both of the synthesized compound and the solvolysed one was confirmed by reversed-phase high pressure liquid chromatography, and 2-dimensional thin-layer chromatography. PMID- 8240987 TI - Heterogeneity in the 5' untranslated region of the rat glucocorticoid receptor mRNA. AB - We have cloned several novel sequences upstream from the first coding exon of the rat glucocorticoid receptor (GR) mRNA using PCR. Analysis of these sequences in RNase protection assays showed that one of the cloned sequences represents the major GR 5' non-coding exon which is expressed in all tissues studied, both at different stages of development and under different hormonal conditions. This major exon is homologous to the human GR 5' untranslated region (UTR). Three other sequences were cloned, but could not be detected in the RNase protection assay, suggesting that they are only minor transcripts, at least under the varying conditions of GR expression studied. One of these sequences is identical to a previously described rat GR cDNA sequence, while another was shown to be contiguous with the rat genomic DNA sequence. PMID- 8240988 TI - New policy alarms French cancer centers. PMID- 8240989 TI - U.S. Congress upholds ban on HIV carriers. PMID- 8240990 TI - Germany plans to restructure medical research. PMID- 8240991 TI - Hodgkin's disease in the setting of HIV infection. PMID- 8240992 TI - p53: a target for new anticancer drugs or a target for old drugs? PMID- 8240993 TI - Leukemia ... the next questions. PMID- 8240994 TI - Hodgkin's disease during HIV1 infection: the French registry experience. French Registry of HIV-associated Tumors. AB - BACKGROUND: The first cases of Hodgkin's disease (HD) associated with HIV infection were reported in 1984. Since then, short series of seropositive patients suffering from HD have been published. In order to identify the characteristics, treatment response and outcome of HIV-associated Hodgkin's disease (HIV-HD), the data of HIV-HD patients recorded between 1987 and 1989 were analysed and compared with those of primary HD patient and with those of HIV associated non-Hodgkin's lymphoma (HIV-NHL), registered during the same period. PATIENTS AND METHODS: The 45 cases of HD collected by the French registry of HIV associated tumors between January 1987 and December 1989 were included in this study. All patients were clinically staged according to the Ann Arbor system. To compare HIV-HD characteristics with those of primary HD, we used a cohort of 407 patients with clinical stages (CS) IA to IVB, who were enrolled between September 1981 and August 1988 in a multicentric clinical trial. To identify the relationship between HIV-HD and the course of HIV infection we studied, when available, the routes of infection, initial CD4 cell count at the moment of HD diagnostic as well as the CDC class of HIV infection and compared these data with the same parameters observed in 142 HIV-NHL enrolled in the registry during the same period. RESULTS: HIV-HD is characterized by an increase in mixed-cellularity histology (49%), with a predominance of advanced stages (75%) and B symptoms (80%). A unique observation is made regarding mediastinal involvement, present in only 13% of HIV-HD (71% in primary HD). The HIV-HD/HIV-NHL ratio was significantly higher in intravenous drug abusers than in male homosexuals. Median CD4 cell count was 306/microliters at HIV-HD diagnosis, and only 11% of the cases were preceded by an AIDS manifestation. With standard therapy, 79% of the patients achieved complete remission, but hematological and infectious complications were very frequent. The progression to AIDS rate was 94% at two years and opportunistic infections were the most frequent cause of death. Overall two-year survival was 41% with 71% for patients with initial CD 4 cell counts higher than 300/microliter and 0% for those with CD4 cell counts lower than 300/microliter (P < 0.01). CONCLUSION: HIV-HD has a particular clinico pathological profile when compared to primary HD, with a predominance of mixed cellularity type, a high frequency of advanced stages and a high proportion of patients without mediastinal involvement. Moreover, HIV-HD seems to occur preferentially in the group of subjects infected by needle sharing. Standard HD therapy seems to be efficient but excessively toxic. PMID- 8240995 TI - Aneuploidy fraction but not DNA index is important for the prognosis of patients with stage I and II breast cancer--10-year results. AB - BACKGROUND: Individual assessment of the prognosis of patients with breast cancer is crucial for the selection of risk-adapted adjuvant therapy and in follow-up. Parameters from DNA flow-cytometry have been shown to provide significant prognostic information, but published results are in conflict and there are only a few investigations with long-term follow-up. The aim of this study is to clarify the impact of tumor DNA data on the clinical course of stage I and stage II breast cancer patients. PATIENTS AND METHODS: Several flow-cytometry DNA analyses were performed on tumor samples derived from 191 breast cancer patients entered in a controlled clinical trial after a median follow-up of more than 10 years. In addition to DNA index (DNI) and the percentage of cells in S phase (SPF), an index, designated aneuploidy fraction (AF), was determined. It ascertains the percentage of aneuploid cells out of all cells in the DNA flow cytometry histogram, and its reproducibility has been tested by measurements of AF in two different samples of the same tumor. Univariate analyses and, in the 122 patients for whom complete information was available, a Cox model, were performed to investigate the individual prognostic impact of flow-cytometry parameters compared with established clinical factors. RESULTS: AF proved to be a very valuable prognostic indicator both in univariate and multivariate analyses, whereas DNI and SPF failed to provide independent prognostic information. The combination of AF and lymph node status clearly identifies different prognostic subgroups in operable breast cancer. CONCLUSIONS: Routine evaluation of patients with breast cancer should include tumor DNA flow-cytometry. Aneuploidy fraction is a valuable tool in assessing an individual patient's prognosis and thus can help in the choice of the appropriate adjuvant treatment strategy. Whether it, rather than DNI and SPF should be used, as we found, needs to be validated in a larger prospective investigation. PMID- 8240996 TI - Cisplatin-etoposide and carboplatin-etoposide induction chemotherapy for good risk patients with germ cell tumors. AB - BACKGROUND: In an attempt to reduce the toxicity of chemotherapy in good-risk testicular cancer patients the two drug combinations, cisplatin plus etoposide (EP) and carboplatin plus etoposide (EC), have been compared. METHODS: Good risk was defined according to the MSKCC and IU criteria. 39 Patients have been treated with EP (cisplatin 20 mg/m2 i.v. and etoposide 100 mg/m2 i.v. on days 1 to 5), and 23 patients received EC (carboplatin 350 mg/m2 on day 1 and etoposide 100 mg/m2 on days 1 to 5). Four cycles of chemotherapy were given at 21- and 28-day intervals, respectively, with delays of up to 7 days in instances of leukocyte counts less than 3.0 x 10(9)/l or platelet counts less than 100 x 10(9)/l. RESULTS: In the EP group 34 (87%) of 39 patients achieved CR (26 with chemotherapy alone, 8 with additional surgery). After a median follow-up of 26 (12-58) months 3 (9%) patients relapsed from CR. Currently 38 patients are alive, and 37 (94%) are NED. In the EC group 20 (87%) of 23 patients achieved CR (15 with chemotherapy alone and 5 with additional surgery). After a median follow-up of 45 (26-57) months 6 (30%) patients relapsed from CR. Currently 19 patients are alive and 17 (74%) are NED. There was no difference in survival between the two groups (p = 0.13), but in the EC group the relapse rate was higher (p = 0.052) and the proportion of patients with NED was lower (p = 0.03) in comparison with EP. Toxicity in both groups was mild and similar, but 3 EP-treated patients presented hair loss. CONCLUSIONS: The study suggests that carboplatin-etoposide combination therapy is inferior to cisplatin-etoposide in patients with good-risk germ cell tumors. PMID- 8240997 TI - Epirubicin and ifosfamide in advanced soft tissue sarcomas. AB - BACKGROUND: To evaluate the feasibility, toxicity and efficacy of the combination of (IFO) ifosfamide and epirubicin (EPI) given at conventional doses for monochemotherapy, we started a phase II study in advanced/metastatic soft tissue sarcoma patients. PATIENTS AND METHODS: Treatment consisted of: epirubicin 75 mg/m2 i.v. day 1; IFO 1.8 g/m2 days 1 to 5; MESNA 20% of the IFO dose at 4-hour intervals three times a day during IFO administration. Cycles were given every 3 4 weeks for at least three cycles. RESULTS: The overall response rate for non visceral sarcomas (51 pts) was 31% (95% confidence limits +/- 13%). Among the 13 visceral sarcomas no response was seen for the leiomyosarcomas of the gastrointestinal tract, whereas one complete and one partial remission were observed for the uterine sarcomas. The duration of response was 10 months (range 5-34+) for complete responses and 9 (range 4-42+) for partial responses. The median survival for responders is 18 months (range 2-60+) and for non-responders 10 months (range 1-33) (p < 0.004). CONCLUSIONS: This combination proved to be feasible and tolerable. The overall response rate does not appear to be superior to those with other standard treatments, but it should be pointed out that our patient population was totally unselected. PMID- 8240998 TI - Phase I and pharmacokinetics study of topotecan, a new topoisomerase I inhibitor. AB - PURPOSE: A phase I study with topotecan (SKF 104864-A, NSC 609699), a semisynthetic analog of camptothecin, was performed using a daily-times-5 regimen, given i.v. q 3 weeks, to evaluate the pharmacokinetics and toxicities of the compound. PATIENTS AND METHODS: Patients with a histologically confirmed diagnosis of a solid tumor no longer amenable to established forms of treatment were eligible for the study. Topotecan was given as a 30 min. infusion daily on 5 successive days, repeated every three weeks. In subsequent patient cohorts the dose was escalated from 0.5 to 1.5 mg/m2/day. Weekly evaluations included hematology and biochemistry. Response to treatment was assessed every 2 cycles. Pharmacokinetics were performed using a HPLC method. RESULTS: Forty-eight patients were entered. The maximal tolerated dose was 1.5 mg/m2/day. The dose limiting toxicity was leucocytopenia. Other major toxicities were alopecia and moderate nausea/vomiting. Partial remissions were observed in one patient with pretreated small-cell lung cancer, one with non-small-cell lung cancer and one with no-pretreated pancreatic cancer, lasting 130-240 days. Pharmacokinetics showed a t 1/2 (alpha) of 8.1 +/- 7.6 min., t 1/2 (beta) of 132 +/- 48 min., Vd,ss 72.7 +/- 26.9 L/m2 and Cltot of 0.57 +/- 0.16 L/min/m2. CONCLUSION: Topotecan is an interesting new topoisomerase I inhibitor exerting antitumor activity in this phase I trial. Leucocytopenia is dose-limiting. The recommended dose for phase II studies is 1.5 mg/m2/day for 5 consecutive days every 3 weeks. PMID- 8240999 TI - Test dose-guided administration of cisplatin in an anephric patient: a case report. AB - BACKGROUND: Although cisplatin pharmacokinetics is well documented in patients with various degrees of renal dysfunction, no information is available concerning cisplatin administration to anephric patients. Since anephric patients may sometimes need cisplatin therapy, it is therefore of importance to define therapeutic guidelines for cisplatin administration in this patient population. PATIENT AND METHODS: Cisplatin was administered to an anephric patient (bilateral nephrectomy) requiring cisplatin therapy for a metastatic carcinoma of the urothelium. A test dose of 12 mg (7.5 mg/m2) of cisplatin was first administered as a 1 hour infusion in order to determine the patient's pharmacokinetic parameters. Filterable and total platinum levels were determined by flameless atomic absorption spectrophotometry. Haemodialysis was started 30 min before the beginning of the cisplatin infusion and was maintained for 4 h thereafter. RESULTS: Under haemodialysis, filterable and total platinum pharmacokinetics after the test dose were comparable with a patient with normal renal function, i.e. with peak plasma concentrations of 126 ng/ml and 166 ng/ml for the filterable and the total platinum, respectively. The area under the curves (AUC) were 154 ng.h/ml for the filterable and 11486 ng.h/ml for the total platinum. The terminal half-lives of filterable and total platinum were 0.42 h and 101 h, respectively. Based on the test dose platinum pharmacokinetics, a therapeutic dose of 100 mg (63 mg/m2) of cisplatin was administered. Following the therapeutic dose, peak plasma concentrations reached 1,120 ng/ml for the filterable and 1,280 ng/ml for the total platinum. The AUCs were 1,609 and 65,556 ng.h/ml for the filterable and the total platinum, respectively, as expected from the predicted AUCs obtained from the test dose pharmacokinetics. The terminal half-lives of filterable and total platinum were similar to the ones observed after the test dose, i.e. 0.36 h and 86 h, respectively. Although the patient died of rapidly progressive hepatic failure, the feasibility of the test dose guided cisplatin administration in an anephric patient is demonstrated. CONCLUSION: This approach may be helpful in monitoring cisplatin therapy in similar cases requiring cisplatin administration. PMID- 8241000 TI - Tamoxifen and hypercalcaemia. AB - The present case demonstrates the importance of close observation, also of serum calcium, when initiating tamoxifen therapy, especially in patients with known osseous metastases. Hypercalcaemia should also be considered as a possible cause of CNS symptoms in patients newly started on endocrine treatment. Hydration with saline is the first step in the management of hypercalcaemia. In patients with a serum calcium level > 3.50 mmol/l (serum ionised calcium approximately 2.00 mmol/l) hydration should be combined with bisphosphonates. PMID- 8241001 TI - Phase II study of continuous intravenous infusion of recombinant interleukin-2 in patients with advanced renal cell carcinoma. AB - BACKGROUND: The goal of this study was to evaluate the therapeutic efficacy and toxicity of recombinant interleukin-2 (rIL-2) administered by continuous intravenous (CIV) infusion to patients with metastatic renal cell cancer (RCC). PATIENTS AND METHODS: Thirty patients with RCC were given rIL-2 18 MIU/m2/d CIV. The schedule consisted of two induction cycles and four maintenance cycles with a 3-week rest period between cycles. Each induction cycle consisted of two infusion periods lasting 120 and 108 hours, respectively, separated by a 6-day rest period. Each maintenance cycle consisted of a 120-hour infusion period. RESULTS: Among 29 assessable patients, the objective response rate was 14% (95% confidence interval, 2% to 26%); one patient achieved a complete response, and 3 partial responses. Ten patients (34%) had stable disease (SD). Median survival was 11 months. Toxicity was generally manageable. Hypotension was universal, but required dose reduction in only 2 patients. Increase in serum creatinine levels was observed in 20 patients, and returned to normal in all but 4 patients after discontinuation of treatment. CONCLUSION: Results confirm the efficacy of rIL-2 in patients with RCC and the feasibility of the treatment in a normal oncology ward. However, responses are observed in a minority of patients, and treatment related toxicity, as well as technical problems, may be troublesome to many patients. PMID- 8241002 TI - A phase II trial of DuP 937 (Teloxantrone) in non-small cell lung cancer. A study of the NCIC Clinical Trials Group. PMID- 8241003 TI - A phase II study of DuP 937 (Teloxantrone) in metastatic malignant melanoma: a study of the National Cancer Institute of Canada Clinical Trials Group (NCICCTG). PMID- 8241004 TI - Etoposide and carboplatin as salvage and first-line therapy in ovarian cancer patients. AB - BACKGROUND: The agents etoposide and carboplatin are active against ovarian cancer and display synergistic anti-tumor activity in animal tumor models. The objective of these two phase II trials was to determine the efficacy and toxicity of the combination of etoposide with carboplatin in previously treated and untreated patients with ovarian cancer. PATIENTS AND METHODS: Etoposide (100 mg/m2) was administered as a one-hour infusion on three consecutive days and carboplatin (400 mg/m2) as a 30-minute infusion on day 2 of each monthly scheduled cycle. In 20 patients, previously treated with cisplatin-containing regimens, a total of 102 cycles was applied as salvage therapy (ST) and in 27 patients, a total of 168 cycles as first-line therapy (FLT). RESULTS: ST yielded 2 complete remissions (CR) and one partial remission (PR); in 7 patients, no evidence of disease (NED) and in 6 patients, no change (NC) were observed. The progression-free intervals (PFI) lasted a median 7.0 months (range < 2-14 months). FLT resulted in 7 CR (4 of them pathologically (p) verified), 11 NED (1 pNED), 3 PR (1 pPR) and 6 NC. The objective response rate was 63% (95% confidence interval: 36-89%). PFI lasted a median 8.0 months (range 3-25+ months); median survival had not been reached at the time of evaluation. Thrombocytopenia (WHO grade 4) was the limiting toxicity. CONCLUSIONS: Although not fulfilling the expectations of synergistic activity as shown in preclinical models, the combination of etoposide with carboplatin is an active and feasible therapy regimen in the out-patient management of ovarian cancer. PMID- 8241005 TI - Cyproterone acetate: hepatotoxicity and prostatic cancer treatment. PMID- 8241006 TI - Approved drugs for unlabeled uses: national health cost containment denies access to state-of-the-art cancer therapies in Italy. PMID- 8241007 TI - Hypertension in the elderly: age- and disease-related complications and therapeutic implications. AB - Effective treatment of hypertension in the elderly requires an understanding of both the progressive course of the disease and the impact of aging on the cardiovascular system, including physiological, genetic, lifestyle, and environmental factors. Review of the literature that has attempted to define the impact of an "aging process" on cardiovascular structure and function reveals a diversity of findings and interpretations. However, in general, normotensive elderly subjects exhibit the heart and vascular characteristics of "muted" hypertension, including many features of younger hypertensive patients: cardiac hypertrophy, diminution in resting left ventricular early diastolic filling rate, increased arterial stiffness and aortic impedance, diminution in the baroreceptor reflex, a diminished response to catecholamines and diminished renal blood flow, and an increase in peripheral vascular resistance (PVR). Treatment of elderly hypertensives is more challenging because of the greater likelihood of the presence of concomitant diseases, most importantly, coronary and peripheral atherosclerosis, renal dysfunction, and diabetes mellitus. Isolated systolic hypertension (ISH), the most common form of hypertension in the elderly, has also been clearly shown to be an important predictor of cardiovascular morbidity and mortality, including coronary artery disease, congestive heart failure, and stroke. Treatment of ISH has been shown to lower systolic pressure safely and effectively in the elderly. By reducing PVR, and possibly the arterial stiffness, and thus the early reflected pulse waves, vasodilators, including calcium antagonists, may lower these three components of arterial impedance, and hence lower the arterial load on the heart. The cardiac hypertrophy and reduced left ventricular filling rate associated with hypertension in older individuals can also be ameliorated, to some extent, by calcium channel blockers. PMID- 8241008 TI - Phosphodiesterase III inhibitors: long-term risks and short-term benefits. AB - Heart failure is now viewed as a disorder of the circulation, not merely the heart, which becomes manifest only when certain compensatory mechanisms break down. After treatment with diuretics, the two main strategies in treating heart failure involve decreasing the work of the heart by vasodilatation or increasing ventricular contractility by positive inotropic agents. It is now apparent, however, that the resulting hemodynamic benefit need not equate with long-term clinical improvement or increased longevity; indeed, the reverse can be true. Inhibitors of phosphodiesterase III, which is specific for the breakdown of cyclic adenosine monophosphate (cAMP), produce useful hemodynamic effects following intravenous and oral dosing, but have not fulfilled their initial promise in the chronic oral treatment of heart failure patients. The reason for reduced survival in the long-term studies of milrinone is not clear, but cardiac arrhythmias, possibly resulting from the increased intracellular levels of cAMP, may be responsible. However, intravenous usage may not suffer from the same limitations as chronic oral dosing. Short-term intravenous administration produces the expected beneficial hemodynamic effects of positive inotropism and vasodilatation. Though infusions of milrinone have been shown to enhance atrioventricular conduction in some, but not all, studies, there appears to be no significant increase in ventricular premature contractions, or ventricular or sustained tachyarrhythmias. Because milrinone does not have a significant adverse effect on His-Purkinje conduction, its use should be well tolerated in patients with intraventricular conduction disturbances. However, accurate assessment of the mortality risk and benefit of short-term intravenous treatment remains to be made in sufficiently powerful prospective, randomized controlled studies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241009 TI - The management of heart failure: a matter of definition? AB - The term heart failure has become a label for more than one clinical entity. For many years heart failure has been used to denote patients with various heart diseases who have begun to suffer from fluid retention, pulmonary venous hypertension, or systemic venous hypertension, either alone or in combination. More recently, the term heart failure has been applied to the combination of effort intolerance and reduced left ventricular contractility due to ischemic heart disease or other myocardial disease. Comparison of the results of epidemiological studies and therapeutic trials is complicated by variation in the composition of the patient populations selected for study. Drug treatment of heart failure remains fairly empirical. Distinction should be made between immediate or prognostic benefits related to the etiological diagnosis, and benefits related specifically to prevention and relief of, for example, fluid retention, rhythm disturbances, or ventricular hypertrophy. The response of individual patients to several forms of drug treatment, including digoxin, ACE inhibitors, and beta-blockade, is unpredictable. Prospective identification of patients liable to respond well to these drugs is not yet possible, but would greatly assist the choice of treatment. At present, trial of therapy is required in each patient to establish benefit and to avoid long-term treatment of nonresponders. PMID- 8241010 TI - Use of milrinone in cardiac surgical patients. AB - Milrinone is shown in 10 patients to be a valuable pharmacological bridge to heart transplantation; it can stabilize and improve decompensated chronic heart failure (CHF) in cases where the response to beta-agonists is inadequate. One patient who had suffered an acute myocardial infarction with heart failure resistant to vasodilators, beta-agonists, and balloon counterpulsation was stabilized with milrinone for 21 days. He was then maintained on ACE inhibitors until heart transplantation 3 months later. The other nine patients with severe decompensated CHF were stabilized on milrinone for between 11 and 51 days. Seven of them received a donor heart. Two patients died of bacteremic shock and terminal heart failure before a suitable organ could be found (31 and 51 days). All patients were clinically improved within 48 hours of the addition of IV milrinone to their therapy. In 55 patients following cardiac surgery, the efficacy and safety of milrinone in the treatment of low cardiac output states is demonstrated. Milrinone has a useful role in the management of patients with circulatory failure both before and after cardiac surgery, and this paper reviews the relevant current literature. PMID- 8241011 TI - Absence of beneficial effect of intravenous metoprolol given during angioplasty in patients with single-vessel coronary artery disease. AB - In a double-blind, randomized, placebo-controlled trial, the possible anti ischemic effect of metoprolol during percutaneous transluminal coronary angioplasty was tested. Electrocardiograms, hemodynamics, and metabolism were studied in 27 patients with a stenosis in the left anterior descending coronary artery. Measurements took place before angioplasty, after each of four 1-minute occlusions and 15 minutes after the last balloon deflation. Patients were randomly given placebo or metoprolol (15 mg as a bolus intravenously, followed by an infusion of 0.04 mg/kg/hr). At the end of the procedure, the rate-pressure product had decreased by 15% (NS) and 23% (p = 0.001) in the placebo and metoprolol groups, respectively, mainly due to similar decreases in heart rate. Metoprolol tended to lower chest pain and reduce precordial ST-segment elevation due to angioplasty, but the effects were not statistically significant. Lactate, hypoxanthine, and urate release immediately after deflation was similar in both groups. Metoprolol reduced arterial plasma hypoxanthine throughout the procedure by about 30% (p < or = 0.02 vs. placebo). Thus, intravenous infusion of metoprolol did not significantly attenuate chest pain and ST-segment elevation, and failed to decrease cardiac lactate and oxypurine release. It did, however, reduce arterial hypoxanthine concentrations during angioplasty, possibly indicating that the beta-blocker inhibits extracardiac ATP catabolism. PMID- 8241012 TI - Risk stratification and prognosis of patients treated with amiodarone for malignant ventricular tachyarrhythmias after myocardial infarction. AB - Seventy-seven consecutive patients (mean age 62 years) with episodes of sustained ventricular tachycardia (VT) or ventricular fibrillation (VF) after acute myocardial infarction (AMI) were evaluated to assess the long-term efficacy of first-line amiodarone treatment and to identify clinical and laboratory factors associated with a high risk of death or arrhythmia recurrence. The presenting arrhythmia was VT in 41 cases (53%) and VF in 36 (47%). VT or VF occurred between the 4th and 90th day after AMI in 45 cases (58%) and later (more than 90 days) in the remaining 32 (42%). The mean number of arrhythmic episodes was 4.2. Forty patients (52%) were in New York Heart Association (NYHA) class I or II, and 37 (48%) were in class III or IV. Mean left ventricular ejection fraction was 32%; ventricular aneurysm was present in 41 subjects. Most patients had multivessel coronary artery disease. Amiodarone was administered as a first-choice drug in all patients, in combination with other antiarrhythmic drugs in 14. By ventricular stimulation after loading doses of amiodarone, sustained VT was inducible in 46 (62%) and noninducible in 28 (38%). During a mean follow-up of 28 months the incidence of cardiac mortality at 1, 3, and 5 years was 21%, 37%, and 47%; of sudden death was 7%, 19%, and 23%; of nonfatal VT recurrence was 13%, 13%, and 24%, respectively. The overall incidence of amiodarone side effects was 35%.2+ was a weak predictor only by univariate analysis (p = 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241013 TI - Nicorandil as a nitrate, and cromakalim as a potassium channel opener, dilate isolated porcine large coronary arteries in an agonist-nonselective manner. AB - Nicorandil is an antianginal vasodilator having a hybrid property between nitrates and potassium channel openers, and cromakalim is a relatively specific potassium channel opener. We investigated whether or not the vasorelaxant actions of the two drugs would be selective for certain vasoconstrictor agonists (simply agonists hereafter), and the underlying mechanisms in isolated porcine large coronary arteries. Both nicorandil and cromakalim produced a complete relaxation in the arteries precontracted with seven agonists, i.e., Bay-K-8644, endothelin, histamine, 5-hydroxytryptamine (5-HT), phenylephrine, PGF2 alpha, and U 46619. The EC50 values (-log M) of nicorandil and cromakalim were 5.20-5.44 and 6.43 6.87, respectively, toward the seven agonists, indicating that the vasorelaxant actions of the two drugs were agonist nonselective. In the arteries precontracted with Bay-K-8644, endothelin, 5-HT, and U 46619, the vasorelaxant action of cromakalim was antagonized by glibenclamide, an antagonist of potassium channel openers, and Schild analysis of these antagonisms yielded pA2 values of 7.10-7.41 for glibenclamide. The vasorelaxant actions of nicorandil in the arteries precontracted with the four agonists each were not antagonized by glibenclamide. Instead, the vasorelaxant action of nicorandil was antagonized by methylene blue (10 microM), an inhibitor of guanylate cyclase, and slightly potentiated by M&B 22,948 (10 microM), an inhibitor of cyclic-GMP phosphodiesterase, in the arteries precontracted with U 46619. These results indicate that the vasorelaxant actions of nicorandil and cromakalim in the porcine large coronary artery are agonist nonselective and that nicorandil exerts such an action entirely as a nitrate, whereas cromakalim does so entirely as a potassium channel opener. PMID- 8241014 TI - Effect of iron overload in the isolated ischemic and reperfused rat heart. AB - It has been suggested that iron might play a pivotal role in the development of reperfusion-induced cellular injury through the activation of oxygen free radical producing reactions. The present study examined the effects of myocardial iron overload on cardiac vulnerability to ischemia and reperfusion. Moreover, the effect of the iron chelator deferoxamine in reversing ischemia-reperfusion injury was studied. Animals were treated with iron dextran solution (i.m. injection, 25 mg every third day during a 5 week period). The control group received the same treatment without iron. Isolated rat hearts were perfused at constant flow (11 ml/min) and subjected to a 15 minute period of global normothermic ischemia followed by reperfusion for 15 minutes. The effects of iron overload were investigated using functional and biochemical parameters, as well as ultrastructural characteristics of the ischemic-reperfused myocardium compared with placebo values. The results suggest that (a) a significant iron overload was obtained in plasma and hepatic and cardiac tissues (x2.5, x16, and x8, respectively) after chronic intramuscular administration of iron dextran (25 mg); (b) during normoxia, iron overload was associated with a slight reduction in cardiac function and an increase in lactate dehydrogenase (LDH) release (x1.5); (c) upon reperfusion, functional recovery was similar whether the heart had been subjected to iron overload or not. However, in the control group left ventricular end-diastolic pressure remained higher than in preischemic conditions, an effect that was not observed in the iron-overloaded group. Moreover, LDH release was markedly increased in the iron-loaded group (x4.2); (d) iron overload was associated with a significant worsening of the structural alterations observed during reperfusion, particularly at the mitochondrial and sarcomere level; (e) after 15 minutes of reperfusion, the activity of the anti-free-radical enzyme, glutathione peroxidase (GPX), was significantly reduced in iron-overloaded hearts, whereas catalase activity was increased; (e) the overall modifications observed in the presence of iron overload were prevented by deferoxamine. In conclusion, this study underlines the possible role of cardiac iron in the development of injury associated with ischemia and reperfusion, and the possible importance of the use of an iron-chelating agent in anti-ischemic therapy. PMID- 8241015 TI - In vitro evaluation of diltiazem on hypothermic injury to immature myocytes. AB - The purpose of the present study was to evaluate the functional and biochemical effects of diltiazem (DTZ) on cardiac myocytes incubated under hypothermic conditions. Cardiac myocytes were isolated from neonatal rat ventricles and cultured for 4 days with MCDB 107 medium. Then, myocytes (12.5 x 10(5) myocytes/flask) were incubated at 4 degrees C for 24 hours in media with or without DTZ at concentrations of 0 M (group C), 10(-7) M (Group D1), 10(-6) M (group D2), 10(-5) M (group D3), or 10(-4) M (group D4). After 24 hours at 4 degrees C, CPK and LDH were measured. The myocytes were then cultured for 24 hours at 37 degrees C to evaluate the recovery of the myocyte beating rate. In group C (n = 7), the recovery ratio of the myocyte beating rate was 29.9% of control (beating rate prior to hypothermic incubation). Groups D1 and D2 (n = 7 each) had approximately the same recovery ratios as group C (24.0% and 24.7%, respectively); however, groups D3 and D4 (n = 7 each) showed no beating rate recovery. Release of CPK and LDH in group C was 112.3 mIU/flask and 457.4 mIU/flask, respectively. Groups D1 and D2 showed no significant differences in both enzymes compared to group C. However, the levels of CPK were significantly higher in group D4 (203.3, p < 0.05), and LDH levels were significantly higher in groups D3 and D4 (669.3, p < 0.05; 883.4, p < 0.02). In conclusion, DTZ showed no protective effects on hypothermic injury to immature cardiac myocytes; moreover, it accelerated cellular injury at the concentrations of 10(-5) and 10(-4) M both functionally and biochemically. Therefore, diltiazem may not be suitable for cardiac preservation during the neonatal period. PMID- 8241016 TI - Hypertensive labeling (reply to Dr. Birkenhager's editorial) PMID- 8241017 TI - Hydrogen peroxide reduces arterial pressure and respiration in the anesthetized dog. PMID- 8241018 TI - Hypertension--a disease or a state of doctors' minds? PMID- 8241019 TI - Negative growth selection against rodent fibroblasts targeted for genetic inhibition of farnesyl transferase. AB - The Ras oncoprotein must be modified by farnesyl transferase (FTase) for biological activity. Therefore, inhibition of FTase may offer a means to block ras induced cell transformation. To address this hypothesis, we have introduced antisense and dominant inhibitory FTase expression plasmids into a panel of normal, mutant ras-, and mos- transformed rodent fibroblasts in an effort to genetically suppress FTase activity. Antisense FTase constructs reduced colony formation efficiency approximately 29% in normal and approximately 41% in ras transformed cells relative to control plasmids. In contrast, antisense FTase plasmids did not exhibit a statistically significant effect on colony formation efficiency in mos-transformed transfectants. FTase alpha N199K is a mutant form of the alpha subunit of FTase that exhibits dominant inhibitory activity versus native FTase. Only mos-transformed transfectants exhibited expression of alpha N199K RNA in 15 of 16 fibroblast lines that were randomly selected and characterized. Our data suggest that genetic inhibition of FTase may result in a selection against animal cell growth. PMID- 8241020 TI - Phorbol esters regulate preprogastrin-releasing peptide messenger RNA in small cell lung cancer cells. AB - The expression of preprogastrin-releasing peptide (GRP) mRNA was studied using human small cell lung cancer (SCLC) cells. By Northern analysis, preproGRP mRNA was stimulated by 4 beta-phorbol 12-myristate 13 alpha-acetate (PMA) in a concentration- and time-dependent manner in these cells. In cell line NCI-H209, the addition of 10(-6) M PMA increased a 0.9-kb mRNA after 8 h. An inactive phorbol ester, 4 alpha-PMA, had little effect on preproGRP mRNA. A nuclear run-on assay indicated that 10(-6) M PMA increased preproGRP transcription 3-fold, whereas beta-actin and glyceraldehyde 3-phosphate dehydrogenase transcription was unaltered. In contrast, PMA had little effect on beta-actin mRNA expression. PMA (1 microM) in the presence of 100 microM 1-(5-isoquinolinesulfonyl)-2 methylpiperazine (H7), a protein kinase C inhibitor, had little effect on preproGRP mRNA. Addition of PMA after protein kinase C down-regulation did not alter preproGRP mRNA. PMA (1 microM) caused translocation of protein kinase C from the cytosol to the membrane of SCLC cells. Also, PMA (10(-6) M) stimulated and H7 (10(-4) M) reduced SCLC growth in vitro. When new synthesis of preproGRP mRNA was blocked by the addition of actinomycin D, preproGRP mRNA remained stable for 15 h. These data suggest that PMA induces transcription of GRP mRNA in SCLC cells. PMID- 8241021 TI - rel/NF-kappa B nuclear complexes that bind kB sites in the murine c-rel promoter are required for constitutive c-rel transcription in B-cells. AB - The c-rel protooncogene, a member of a transcription factor family that includes NF-kappa B, displays a complex pattern of gene expression. To understand the basis of this expression, the regulatory region upstream of the murine c-rel transcription start sites has been cloned and characterized. Transcription of the murine c-rel gene initiates at multiple sites downstream of a GC-rich region conserved in the chicken c-rel promoter. This conserved region contains consensus transcription factor binding sites for SP-1 and NF-kappa B (kB3 site) and is sufficient for basal expression in Jurkat T-cells. In contrast, two additional NF kappa B-like sites (kB1 and kB2) and an octamer consensus binding site, all located upstream of the conserved region, are required for expression of promoter reporter gene constructs in the B-cell line I29B. NF-kappa B sites kB1 and kB3 bind p50/65 and p50 homodimers, whereas kB2 binds a distinct complex. The consensus octamer site, although only able to bind Oct1 and Oct2 with low affinity, appears to overlap with a binding site for a novel protein(s) expressed in I29B cells. Cotransfection studies show that p75-c-rel and a carboxyl-terminal truncated c-rel protein that lacks the known trans-activating domain both up regulate the c-rel promoter in I29B cells via a mechanism independent of the NF kappa B motifs, whereas a mutant c-rel protein lacking the DNA binding domain has no effect. Together, these findings suggest that, in this B-cell line, trans activation of the c-rel promoter by rel proteins is via an indirect mechanism. PMID- 8241022 TI - Changes in ribosomal protein and ribosomal RNA synthesis during rat intestinal differentiation. AB - Subtraction hybridization studies, used to identify genes involved in the control of enterocyte proliferation and/or differentiation, allowed detection of a clone shown to have homologies with rat, chicken, and human acidic ribosomal phosphoprotein P1. Since increases in P1 transcript have been associated with intestinal malignancy, we explored the relationship of P1 and other ribosomal proteins to normal intestinal proliferation and differentiation. Male rats were used to prepare enterocytes as isolated cell fractions representative of the crypt to villus axis of differentiation. Total RNA was extracted from pooled cell fractions and evaluated for mRNA and rRNA steady-state levels. Nuclei were prepared from isolated enterocytes, and nuclear runoff studies were performed to estimate rates of nascent transcription. The P1 complementary DNA from the crypt cell library detected a mRNA of 650 base pairs which showed approximately 8-fold greater steady-state levels in crypt than in villus cells. Similar crypt specificity was also noted for mRNAs coding for elongation factor EF-12 and for ribosomal proteins P0, P1, and S6 (using clones from Y-L. Chan and I. G. Wool). In contrast, 28S rRNA steady-state levels did not differ between villus and crypt, indicating that ribosomal content had remained constant. In situ hybridization studies confirmed the predominant crypt localization of P1 mRNA. Nascent transcription rate studies showed that the proportion of newly synthesized P1 mRNA to total RNA was the same for the villus and crypt, suggesting that the lower content of villus P1 mRNA may be due to increased degradation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241023 TI - Serum deprivation response gene is induced by serum starvation but not by contact inhibition. AB - The relationship between quiescence induced by serum deprivation and that induced by high cell density (contact inhibition) is still unclear. Here, we describe a gene, named sdr (serum deprivation response), whose mRNA level is highly induced in serum starved but not in density dependent growth arrested NIH3T3 cells. sdr induction seems to be coupled to growth arrest since it does not become apparent when transformed NIH3T3 cells are cultured in low serum. The expression of sdr is down-regulated within 6 h after the addition of serum or epidermal growth factor to serum starved cells. Although a transient reinduction is noticed at later times after fetal calf serum stimulation, this is not the case for epidermal growth factor, for which the sdr mRNA level remains down-regulated. The data presented here point to the possibility that the density dependent growth arrest state cannot be simply explained by local growth factor depletion, as occurs in overcrowded cultures, but it could be the consequence of a more complex pathway mediated by cellular interactions. PMID- 8241024 TI - Chimeras of herpes simplex viral VP16 and jun are oncogenic. AB - The Jun protein binds DNA and regulates transcription as a component of the AP-1 transcription factor complex. In its oncogenic form, Jun can transform cells in culture and cause tumors in animals. Both trans-activation and transformation require several functional domains of Jun, including an amino-terminal trans activation domain. In this study, properties of Jun required for trans-activation and transformation were explored by replacing the trans-activation domains of c Jun and its oncogenic counterpart, v-Jun, with the constitutively active trans activation domain from the herpes simplex virus VP16 protein. The VP16-v-Jun chimera retained similar oncogenic properties to its parent, v-Jun. The VP16-c Jun chimera, however, was considerably more oncogenic than c-Jun. Substitutions of a phenylalanine in the VP16 domain of the VP16-c-Jun chimera diminished or abolished transformation. Each of the chimeras bound to the AP-1 consensus recognition sequence from the collagenase promoter or from the human T-cell leukemia virus type I long terminal repeat in vitro. None of the VP16-Jun chimeras efficiently stimulated transcription from the collagenase promoter or an artificial promoter containing the human T-cell leukemia virus type I element in vivo. These results demonstrate that the Jun trans-activation domain can be replaced by a heterologous trans-activation domain with retention of oncogenic activity. However, this oncogenic activity is not reflected in the trans activating properties of the chimeras. PMID- 8241025 TI - Retroviral gene transfer of epidermal growth factor receptor into HL60 cells results in a partial block of retinoic acid-induced granulocytic differentiation. AB - HL60 cells are devoid of endogenous epidermal growth factor receptor (EGFR). They respond to retinoic acid and undergo terminal granulocytic differentiation. EGFR complementary DNA was introduced into HL60 cells by retroviral gene transfer. Scatchard plot showed that the binding characteristics are identical to those of A431 cells. HL60-EGFR cells were estimated to express 34,000 EGFR/cell (Kd = 5 nM). The tyrosine phosphorylation upon ligand binding is the first step of signal transduction. The dominant phosphotyrosyl proteins in epidermal growth factor stimulated HL60-EGFR cells include a 170 kDa protein (EGFR itself), and 125 and 53 kDa proteins. The EGFR signal results in the induction of 92 kDa gelatinase/matrix metalloproteinase in HL60-EGFR cells, thereby providing evidence of the function of the exogenous EGFR and a semiquantitative measure of the EGFR signal. These HL60-EGFR cells offer a unique opportunity to examine the potentially important role of EGFR (c-erbB) in maintaining homeostasis between self-renewal and differentiation. c-erbB has been shown to play a physiological role in the self-renewal of the very early avian stem cells which do express EGFR. The v-erbB (double truncated EGFR) has been shown to cause avian erythroblastosis. We found that these HL60-EGFR cells responded to retinoic acid differently from the HL60-control cells. A partial block of only 45% granulocytic differentiation and concomitant proliferation was noted, consistent with a shift of balance between self-renewal and differentiation toward the former. PMID- 8241026 TI - Identification of growth inhibited cells by retention of a lipophilic fluorescent dye. AB - Cellular proliferation is regulated in both positive and negative ways. However, direct selection for growth inhibitory control elements is limited by the difficulty in identifying a growth inhibited cell against a background of cells which are proliferating. This study describes a positive selection technique for growth inhibited cells. This method is based on the retention of a lipophilic fluorescent dye which nonspecifically labels plasma membranes and distributes between daughter cells with membrane lipid as cells proliferate. Characterization of this assay is described using an epithelial cell line which is growth inhibited in response to transforming growth factor beta (TGF-beta) and dexamethasone and several mutant clones of that line which lack responsiveness to TGF-beta. Retention of dye in response to the growth inhibitors is proportional to the inhibition of thymidine incorporation of those cells. Mixing experiments were also carried out in which G418 resistant TGF-beta responsive epithelial cells were mixed with TGF-beta nonresponsive mutants. The mixture was labeled with PKH-2 and exposed to TGF-beta for 3 days. Subsequently, consecutive fractions of cells sorted on the basis of fluorescence intensity were selected in G418, and the TGF-beta responsive epithelial cells were found predominantly in the most fluorescent cells in the population. This method provides a positive selection for growth inhibited cells which may, in combination with classical gene transfer techniques, provide a way to select for growth inhibitory genes in a manner analogous to the focus forming assay selection for oncogenes. PMID- 8241027 TI - Colon carcinoma cells blocked in polarization exhibit increased expression of carcinoembryonic antigen. AB - Carcinoembryonic antigen (CEA) is an oncofetal protein whose regulation is poorly understood, although CEA is commonly expressed on many carcinoma cell types and enhances experimental metastases. The abundance of membrane-associated CEA was increased 3-fold when HD6 colon carcinoma cells were prevented from polarizing by culture for 3 days in low calcium medium. Polarization is an early event in HD6 cell differentiation, with the polarized cells forming a tight, laterally adherent monolayer by culture in normal calcium medium. Lateral adherence can occur because 3 days of culture in normal calcium medium increases expression of calcium-dependent intercellular adhesion proteins: a 35-fold increase in membrane abundance of LCAM and a 16-fold increase in membrane abundance of the desmosomal protein desmoglein I. Polarized HD6 cells exhibit low levels of CEA only at their apical luminal surface. Rounded, unpolarized HD6 cells do not exhibit increases in either LCAM or desmoglein I membrane expression, but express increased levels of CEA molecules throughout their cell surface, where they act as intercellular adhesion molecules, allowing unpolarized cells to form random cell to cell contacts. Cells cultured in low calcium medium form calcium-independent cell aggregates whose formation can be blocked by Fab' fragments of anti-CEA monoclonal antibody col-1. The familiar pattern of random, multilayered associations of tumor cells both in vitro and in xenographs in vivo may be due to intercellular adhesion mediated by CEA which is up-regulated and expressed throughout the cell surface of unpolarized tumor cells. PMID- 8241028 TI - Time course changes in cytoskeletal structures of cultured endothelial cells exposed to shear stress. AB - Changes in patterns of microfilament distribution were analyzed using cultured porcine aortic endothelial cells under shear flow. Within the elongated endothelial cells after shear flow exposure (20 dyne/cm2, 0-24 h), F-actin filaments were mainly observed as stress fibers. After 24 h exposure, the fluorescence intensity of rhodamine-phalloidin-stained cells was enhanced, indicating the increase in F-actin content. It was suggested that not only F actin distribution but also the content may be affected by shear stress at the early stage of exposure. PMID- 8241029 TI - The importance of taper proximal and distal to Y-bifurcations in arteries. AB - Changes in luminal cross-sectional area from human cerebral arteries, porcine renal arteries and human aorto-iliac junctions were found to be both significant and gradual. Proximal to the flow divider, luminal cross-sectional area increased by 10.8 +/- 5.7% for the human aorto-iliac and 94.5 +/- 5.0% for the cerebrals. Distal to the flow divider, the summed luminal area of the two daughter branches decreased by 36.4 +/- 4.0% for the cerebrals and 15.9 +/- 5.2% for the renals. Taper was defined as the ratio of the area change to the vessel length over which the change occurred. Positive taper corresponded to a narrowing in cross-section while negative taper represented an expansion in luminal area. All taper values were significantly different from zero (t-test, P < 0.05). Arterial ellipticity was examined from the ratio of lateral and antero-posterior diameters of cross sections in the parent trunk from human aorto-iliacs and porcine renal arteries. We have shown that ellipticity begins one parent-tube diameter from the apex of the bifurcation in both samples studied. These large changes in luminal area and taper, which have been overlooked previously, have significant implications for the propagation of flow and pressure waves as well as for flow patterns. PMID- 8241030 TI - A computational fluid mechanical study of blood flow in a variety of asymmetric arterial bifurcations. AB - In the present study, the results of an attempt to estimate the flow field at arterial bifurcations are shown and discussed with respect to the pathogenesis of vascular disorders. The Navier-Stokes equations for incompressible Newtonian fluid flow were solved using a finite volume method with a body-fitted coordinate (BFC) system. The arterial models had various branching angles and mother daughter diameter ratios. The boundary conditions specified are: steady and pulsatile fully developed flow at the inflow cross-section and a non-slip condition at the rigid wall. Results obtained were displayed using color computer graphics and an animation technique. Complex vortices were always found at the bifurcation regardless of the branching angle. At the proximal region of the bifurcation, where the early atherosclerotic plaque usually develops, strong unsteady vortices were found to access the wall in the diastolic phase. Therefore it is necessary to take them into account when discussing the development of atherosclerosis. PMID- 8241031 TI - The role of vascular smooth muscle cell phenotypic modulation at the aortic branch in atherogenesis. AB - To elucidate the mechanism of the development of atherosclerosis, we have investigated the cell population of phenotypes of contractile (C-SMC) and synthetic (S-SMC) states of SMCs at proximal and distal areas of bifurcation of the celiac and superior mesenteric arteries in children and young persons by transmission electron microscopy. The previous studies in patients with hypercholesterolemia and who were young indicated that percentages of proximal area at bifurcation of both arteries were greater than that of C-SMC (P 0.01), and that C-SMCs at distal area were less than that of S-SMC (P 0.05). Ultrastructurally, SMCs at proximal area were S-SMCs containing many synthetic organelles and intermediate filaments. On the other hand, those at distal area were C-SMCs containing actin, myosin, dense bodies and microtubules. In the present study, we have ascertained that the phenotypic modulation of SMCs in the intima and media might correlate to the physico-medial relationship between SMCs and elastic tissues. In this communication, we have observed that the intimal SMCs transformed their phenotypes from the internal elastic layer (S-SMC) to the superficial layer (C-SMC), and that the medial SMCs in the arteries of the young clearly consisted of two types: one type adhered to the elastic layer and the other type existed with the separated one. The difference between both areas in the relatively young need to be observed in detail from now on. In summary, the vascular SMCs and the elastic lamina are considered to contribute to the subsequent phenotypic modulation and their migration of SMCs. PMID- 8241032 TI - Market is open for SQUID magnetoencephalography. PMID- 8241033 TI - Morphological responses of single endothelial cells exposed to physiological levels of fluid shear stress. AB - To investigate the basic processes underlying flow induced morphological changes of vascular endothelial cells (ECs), we have examined the responses of single ECs to fluid mechanical forces under controlled flow conditions in vitro. We made a parallel plate culture flow chamber which could be mounted on an inverted microscope equipped with various optical systems. Video-enhanced microscopic images were recorded with a time-lapse, laser video-disk recorder and then analyzed frame by frame. In contrast to ECs in a confluent monolayer, ECs in sparse culture did not show a parallel alignment to the flow direction even after a 100 h exposure to flow at a wall shear stress level of 10 dyne/cm2. However, rapidly after changing from a weak shear stress (1 dyne/cm2 or less) to a high shear stress (6 dyne/cm2 or more), sparsely cultured ECs extended lamellipodia predominantly on the downstream side and began to migrate preferentially downstream. If the viscosity of the circulating medium was raised by adding a small amount of methyl cellulose or polyvinylpyrrolidone, ECs exhibited these characteristic morphological changes under low wall shear rates (100 s-1 or less). We also found that these changes required Ca2+ in the circulating medium. These results indicate that single ECs can respond to flow in a rapid, shear stress and Ca2+ dependent manner. PMID- 8241034 TI - Finite element analysis in three-dimensional flow through a lateral saccular aneurysm. AB - The three-dimensional flow pattern in a lateral saccular aneurysm was investigated by a finite element method. The aneurysm model had a dome-shaped diverticulum arising from one side of the straight circular tube. The steady Navier-Stokes equations and the equation of continuity were applied. The finite element equations were derived from the Galerkin process with each hexahedral element, in which velocities were approximated by quadratic polynomials and pressures by linear polynomials. The non-linear finite element equations were solved by a Newton-Raphson method. Calculations were performed of the steady flow at various Reynolds numbers. The conclusions for flow patterns with a Reynolds number of 1000 is as follows: (i) a separation vortex occurred in the whole circumferential region of the aneurysm, (ii) the separation vortex was distorted by the geometrical shape of the aneurysm, and (iii) two other vortexes induced by the separation vortex existed at the proximal and the distal wall of the aneurysm. These disturbances of flow through an aneurysm are thought to play a significant role in the formation of thrombi and the rupture of an aneurysm. PMID- 8241035 TI - Influence of shear stress on endothelial cell shapes and junction complexes at flow dividers of aortic bifurcations in cholesterol-fed rabbits. AB - We studied the effects of hemodynamic forces on endothelial cell shape and junction complexes in sudanophilic and non-sudanophilic regions at flow dividers of aortic bifurcations in normolipidemic and hyperlipidemic rabbits by scanning electron microscopy (SEM) and the freeze fracture method. Flow profiles and shear stresses were obtained by tracing the paths of particles in a transparent vessel in steady flow. No apparent differences between normolipidemic and hyperlipidemic rabbits were observed by SEM. The tip of the apex of flow dividers, a point of stagnated flow, and the leading edge, which was exposed to laminar, high shear stress, were non-sudanophilic, and were covered by, respectively, round and long fusiform endothelial cells. The tight junctions were continuous and gap junctions had a regular shape. The hip of the flow dividers of branchings, a relatively low shear stress region, was sudanophilic and was covered by ellipsoidal cells with discontinuous tight junctions and irregularly shaped gap junctions. Our results suggest that endothelial cells exposed to relatively low wall shear stress, but not to high shear stress, may be functionally activated, leading to an increased intercellular permeability, which may make these regions vulnerable to atherosclerosis. PMID- 8241036 TI - International photorefractive keratectomy market thrives while FDA stalls. PMID- 8241037 TI - Intraoperative intraocular endoscopy in transscleral suture fixation of posterior chamber lenses: consequences for suture technique, implantation procedure, and choice of PCL design. AB - BACKGROUND: Transscleral suture fixation of posterior chamber lenses (PCLs) in the absence of capsular support causes minimal long-term alteration of the blood aqueous barrier, if two requirements are fulfilled: 1) the transscleral suture has to penetrate exactly through the ciliary sulcus, and 2) the PCL haptics have to be directed into the sulcus and secured there. METHODS: The surgical results of our standard techniques were controlled intraoperatively by means of intraocular endoscopy in every patient since May 1991. Different alterations were necessary to improve the incidence of sulcus penetration and implantation. RESULTS: With our conventional suture techniques, the needle penetrated the ciliary processes in the majority of eyes. Best results were achieved by passing the needle from the outside into the eye before opening the globe. When the eye was already hypotonic, the ciliary processes tended to prolapse in front of the needle tip, resulting in pars plicata fixation. With perforating keratoplasty, passing the needle from the inside out by feeding one's way into the sulcus with the needle tip gave good results. Even correct needle penetration through the sulcus did not guarantee correct positioning of the PCL haptics in the sulcus. CONCLUSIONS: Selecting a suitable PCL design and a new implantation technique which reduces the angle of PCL implantation, the rate of correctly positioned PCLs in transscleral suture fixation is increased considerably. PMID- 8241038 TI - Effects of air injection into the corneal stroma of man: an in vitro study. AB - BACKGROUND: Corneal stromal air injection has been described for assisting the deep dissection of the recipient corneal bed in lamellar grafting. The consistent finding of air entering the anterior chamber is considered an undesirable complication of the procedure. The route of air entry into the anterior chamber has not been investigated previously. METHODS: The route of air passage through the cornea was studied in 10 human cadaver eyes by direct viewing of the inner corneal surface with an operating microscope and by electron microscopy. RESULTS: Air entered the anterior chamber peripheral to Descemet's membrane in every case in the absence of pre-existing corneal pathology. The air expansion of the cornea occurred only along planes occupied by keratocytes and their cell processes and this expansion caused disruption of the keratocytes. CONCLUSIONS: Air injection into the cornea caused bubbles to enter the anterior chamber peripheral to Schwalbe's line, through the trabecular meshwork. The air produced cleavage of the collagen lamellae along those planes occupied by keratocytes and their cell processes since this provides a path of least resistance. Disruption of the keratocytes is a direct mechanical effect. Descemet's membrane and Bowman's layer act as effective barriers to air passage in the normal cornea. PMID- 8241039 TI - Computer-assisted corneal topography: accuracy and reproducibility of the topographic modeling system. AB - BACKGROUND: Assessment of the efficacy and stability of novel refractive surgery techniques and comparative studies require accurate and reproducible corneal mapping technology, ideally to +/- 0.25 D. A vertically mounted Topographic Modeling System (TMS) system was evaluated. MATERIALS AND METHODS: Six calibrated PMMA spheres (30.00, 35.00, 43.00, 50.00, 55.00, and 60.00 D) were used. Two examiners performed three measurements on each test sphere and compared appropriate data with a previously tested and calibrated digital auto-keratometer (SK-1, precision < 0.25 D). The accuracy and reproducibility of all 25 rings of the TMS were studied as a function of focus (250 to 500 microns), centering (100 to 300 microns), and integrity of the inner and outer reflected placido images to mimic tear breaks at the corneal apex and peripheral vignetting and shadows produced by eyelids and eyelashes. RESULTS: The accuracy of the 25 rings decreased from the center to the periphery. The deviation scores (mean difference of three readings between measured and calibrated surface powers) of rings 3 through 25 were 100% within +/- 0.25 D only for the 43.00-diopter sphere. They were 56%, 68%, 43%, 13%, and 17% within +/- 0.25 D respectively for the 30.00-, 35.00-, 50.00-, 55.00-, and 60.00-diopter spheres. For 500 microns of defocusing, we observed a mean deviation of 0.35 D for the flattest sphere to 2.30 D for the 60.00-diopter sphere. For the same amount of defocusing, the error produced by the TMS increased progressively with the sphere dioptric power. Masking the first four and six rings, one by one, showed topographic maps to be valid only when all four central placido images were fully read by the computer. CONCLUSIONS: The TMS clinical performance could be improved by 1) increasing focus depth of field, 2) blocking data processing when central rings are incomplete or missing, 3) increasing the accuracy over the 30.00- to 60.00-diopter range, and 4) providing the user with calibration spheres. PMID- 8241040 TI - Influence of radial keratotomy on endogenous hyaluronan in cornea and aqueous humour. AB - BACKGROUND: Hyaluronan is known to appear in corneal wound tissue and is probably involved in the healing process. We measured the changes of endogenous hyaluronan in rabbit cornea and aqueous after radial keratotomy. METHODS: Corneal and aqueous hyaluronan concentrations were measured at different time intervals after radial keratotomies. Hyaluronan was extracted from corneal tissue with two methods employing either pronase or NaCl and measured with a radioimmunoassay. Histochemical staining for hyaluronan was done on the operated eyes. RESULTS: Changes in corneal hyaluronan showed a similar time course with either extraction method. Maximal concentration was reached after 7 to 14 days with a return to normal values in 60 days. Aqueous hyaluronan and proteins were not significantly altered after surgery. Histochemical staining of the corneas demonstrated hyaluronan around the incisions along their entire extent. Blood vessels were absent in the wounds. An increase in elongated fibroblast-like cells was seen in the wound area. CONCLUSIONS: Both quantitatively and histochemically, there is a pronounced increase of corneal hyaluronan in avascular, radial keratomy wounds in the rabbit. The findings suggest a role of hyaluronan in corneal wound healing. PMID- 8241041 TI - Model for predicting the optical performance of the eye in refractive surgery. AB - BACKGROUND: Predicting the outcome of refractive surgical procedures is one of the chief goals of the refractive surgeon. Besides perfecting the change required to nullify a refractive error, the optical quality of the postsurgical eye should be maximized. METHODS: A model eye useful for predicting the likely effects on the optical performance of the eye as a result of refractive surgical intervention is presented. The model features aspheric ocular interfaces, a gradient refractive index within the lens, and a uni-index cornea. The dimensions of the model parameters are taken from the literature. Baker's method of ray tracing through aspheric surfaces is used to predict the lateral spherical aberration of the eye for specific pupil sizes. The results for various operational conditions such as number of lens layers and corneal shape were calculated. RESULTS: The model predicts that optimal optical imagery is produced when the corneal profile is represented by a flattening ellipse (shape factor = .65 to .85). Ideally, in refractive surgery involving the cornea, the postoperative corneal contour should conform to this flattening ellipse. PMID- 8241042 TI - Corticosteroids vs diclofenac in the treatment of delayed regression after myopic photorefractive keratectomy. PMID- 8241043 TI - Assessing the efficacy of topical corticosteroids following radial keratotomy. AB - BACKGROUND: The postoperative use of topical corticosteroids may decrease the risk for regression and thereby improve the refractive outcome following radial keratotomy. METHODS: Twenty patients with identical preoperative refractions in each eye underwent bilateral radial keratotomy in one sitting. One eye was treated with dexamethasone twice daily for 3 months postoperatively; the fellow eye was not. RESULTS: Three months postoperatively, there was no difference between the refractions of the two groups of eyes. CONCLUSION: The use of topical dexamethasone after radial keratotomy for 3 months does not improve the refractive outcome. PMID- 8241044 TI - Keratoconus and corneal hydrops associated with compulsive eye rubbing. PMID- 8241045 TI - High-frequency ultrasound measurement of the thickness of the corneal epithelium. PMID- 8241046 TI - Arcuate keratotomy for the correction of spherical hyperopia in human cadaver eyes. AB - BACKGROUND: A new experimental surgical technique to correct spherical hyperopia by steepening of the central corneal curvature was performed on human cadaver eyes. METHODS: Ten eyes were used in the study. All were pretreated with glycerin to ensure a uniform corneal thickness between .55 and .65 mm on ultrasonic pachometry. A constant intraocular pressure of approximately 30 to 40 mm Hg was maintained in each eye by the injection of saline into the vitreous cavity. This was verified by pneumotonometry. A vertical blade diamond knife was set at 100% of the thinnest of four paracentral readings. Each eye underwent preoperative computed topography. One set of five eyes had 12 incisions made following a 5.75 millimeter diameter Mendez hexagonal marker that included unconnected T incisions at each junction (ie, "Hex T" pattern). The other set of five eyes had four arcuate incisions made following a 6-millimeter diameter zone marker; each incision was 60 degrees in arc. Immediately after surgery, computed topography was repeated. RESULTS: The hexagonal keratotomy set of eyes had an average steepening of the cornea of 0.80 D with a range of -1.05 to + 4.38 D. The arcuate keratotomy set had an average steepening of +2.12 D with a range of +1.27 to +3.27 D. CONCLUSIONS: This study suggests arcuate keratotomy may be a more effective procedure in the correction of spherical hyperopia when compared with hexagonal keratotomy. The amount of corneal steepening achieved in a cadaver eye model may not be the same as when performed in vivo. PMID- 8241047 TI - Superior keratoconus. PMID- 8241048 TI - Laser epithelium removal before photorefractive keratectomy. PMID- 8241049 TI - Central steep islands immediately following excimer photorefractive keratectomy for myopia. PMID- 8241050 TI - A high frequency of hybridomas from M54 mu heavy chain transgenic mice initially co-express transgenic and rearranged endogenous mu genes. AB - The M54 transgenic mouse line, which carries the 17.2.25 Ig mu heavy chain gene, rearranges Ig heavy chains and expresses both transgenic and endogenous mu. B cell lineage development is selectively impaired in these mice and cells that simultaneously express transgenic and endogenous mu ('double-producers') are common amongst the B cells and plasma cells that do develop. Weaver, Imanishi kari, Baltimore and colleagues failed to obtain double-producing hybridomas from M54 mice; however, molecular and serologic studies presented here show that such hybridomas are readily generated. These hybridomas are extremely unstable and rapidly yield variants producing either transgenic or endogenous mu. Therefore the stable cloned lines we obtained, like Weaver et al., were almost all single or non-producers. We also found that the VH gene usage in our hybridomas was skewed towards the JH proximal (VHQ52, VH81X) families, supporting the idea that the expression of the M54 transgene alters the endogenous Ig repertoire. PMID- 8241051 TI - The role of macrophages in antigen presentation and T cell tolerance. AB - Bone marrow derived cells (dendritic cells, macrophages and B cells) are involved in antigen presentation and T cell tolerance. However, the precise functions of each cell type remain unclear. To determine the role of macrophages we produced transgenic mice expressing I-E molecules only on macrophages, by introducing the hybrid gene containing the colony stimulating factor-1 (CSF-1) receptor promoter region and the structural gene encoding E alpha d into C57BL/6 mice. In these mice I-E restricted antigen presentation and T cell priming were impaired. With respect to T cell tolerance, I-E reactive T cells were anergized but not clonally deleted. These results clearly demonstrate that macrophages by themselves are defective in efficient I-E restricted antigen presentation, so that T cells exposed to antigens expressed on macrophages are led to anergy. PMID- 8241052 TI - Alterations in gene expression associated with stepwise acquisition of malignancy in murine cytotoxic T cell lines. AB - We have isolated a series of variant cell lines from a murine CD8+ T cell clone representing distinct stages in stepwise acquisition of malignancy. A first type of variant has acquired independency of restimulation with MHC/Ag but has kept dependence on IL-2 for continuous growth in culture. A second type of variant has acquired, in addition, independency of IL-2. A third type of variant was isolated from tumors induced upon injection of IL-2 independent variants into syngeneic mice. Clonal relatedness between the cell line was ascertained by Southern blot and sequence analyses of their TCR beta chain genes. The cell lines were analyzed for their expression of genes typical for CD8+ T cells, using Northern blot hybridization, flow cytometry, and functional methods. Concentrating on the transition from IL-2 dependent to IL-2 independent cellular growth, we find the same triad of changes in two independently derived groups of variant cell lines: loss of expression of the CD8 alpha gene with concomitant loss of CD8 from the cell surface, a slight but significant overexpression of IL-2R alpha and beta chains with increased low affinity IL-2 binding sites, and constitutive overexpression of c-myc. Autocrine IL-2 dependent growth could be excluded. Expression of p56lck did not vary between the cell lines. We discuss the possibility that IL-2 independent growth may be associated with intracellular redistribution of p56lck from CD8 alpha to IL-2R beta, thus generating constitutively active IL-2R. Ex vivo established tumor variants differed from their parental culture cell lines by their constitutive secretion of IFN gamma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241053 TI - ER-MP12 antigen, a new cell surface marker on mouse bone marrow cells with thymus repopulating ability: I. Intrathymic repopulating ability of ER-MP12-positive bone marrow cells. AB - We searched for new cell surface markers that allow a positive identification of thymus-repopulating cells in the bone marrow (BM) of the mouse. Recently we raised two rat monoclonal antibodies (ER-MP12 and ER-MP20) that recognize cell surface antigens expressed by mouse haematopoietic progenitor cells, among which are progenitor cells of the macrophage lineage. Here we show that the ER-MP12 antigen, but not the ER-MP20 antigen, is also expressed by BM cells with thymus repopulating ability. Using ER-MP12 and ER-MP20 in two-colour immunofluorescence analysis six subpopulations of BM cells can be identified. The thymus repopulating ability of each BM subpopulation was assessed after fluorescence activated cell sorting and subsequent intrathymic injection into sublethally irradiated Thy-1 congenic recipient mice. Thymus-repopulating activity appeared to be exclusively confined to two subsets of BM cells expressing either high or intermediate levels of the ER-MP12 antigen, but lacking ER-MP20 antigen expression. These BM subsets comprised 1-2% and 30% of total nucleated BM cells respectively. The frequency of thymus-repopulating cells was maximal in the minor BM subpopulation with the highest level of ER-MP12 antigen expression. We conclude that ER-MP12 detects a hitherto unknown cell surface marker expressed by BM cells with thymus-repopulating ability. PMID- 8241054 TI - ER-MP12 antigen, a new cell surface marker on mouse bone marrow cells with thymus repopulating ability: II. Thymus-homing ability and phenotypic characterization of ER-MP12-positive bone marrow cells. AB - In the accompanying paper we showed that six distinct subsets of bone marrow (BM) cells can be identified using the mAb ER-MP12 and ER-MP20 in two-colour immunofluorescence analysis. Upon intrathymic transfer into sublethally irradiated mice thymus-repopulating ability was restricted to ER-MP20- BM cells expressing either high or intermediate levels of the ER-MP12 antigen (1-2% and approximately 30% of BM nucleated cells respectively). The highest frequency of thymus-repopulating cells was found in the minor subset of ER-MP12(+)+20- BM cells. In the present study we demonstrate that upon intravenous transfer, thymus homing and -repopulating BM cells are exclusively confined to the ER-MP12(+)+20- and ER-MP12+20- subpopulations, the highest frequency being detected among ER MP12(+)+20- BM cells. Analysis of the peripheral blood leucocytes of reconstituted mice showed that not only prothymocytes but also progenitor cells of the B cell lineage as well as the myeloid lineage were present within both subsets. Three-colour flow cytometric analysis revealed that ER-MP12(+)+20- BM cells in particular were phenotypically heterogeneous with respect to the expression of the cell surface markers Thy-1, Sca-1, CD44, B220 and c-kit. Taken together our data demonstrate that ER-MP12 positively identifies BM cells with the ability to home to and repopulate the thymus. The phenotypic heterogeneity displayed by the ER-MP12(+)+20- BM subset, containing the highest frequency of thymus-homing and -repopulating cells, provides a basis for further separation of prothymocyte activity from other haematopoietic activities in the BM of the mouse. PMID- 8241055 TI - Role of conserved regions of class I MHC molecules in the activation of CD8+ cytotoxic T lymphocytes by peptide and purified cell-free class I molecules. AB - To analyze the molecular interactions involved in CD8+ cytotoxic T lymphocyte (CTL) recognition quantitatively, we developed a cell-free antigen presenting system. Genetically engineered soluble H-2Dd molecules coated on plastic microtiter plates could present HIV envelope peptide to an antigen-specific CTL clone, inducing it to produce IFN-gamma in the absence of accessory cells and their accessory or co-stimulatory molecules. The peptide-MHC complexes were functionally stable for over 24 h. The magnitude of T cell activation was dependent on the concentrations of both class I MHC molecule and the peptide, but was more sensitive to the concentration of the MHC molecule than to that of peptide. This result suggests that one MHC molecule can play more than one role in activating the CTL. One such role is the interaction between CD8 and a conserved region of class I MHC, as suggested by the finding that holding the total MHC concentration constant with an irrelevant class I MHC molecule (H-2Kb engineered to have the same alpha 3 domain as H-2Dd) made the T cell response less sensitive to the change in concentration of the relevant MHC molecule (H 2Dd). The irrelevant class I MHC molecule (H-2Kb), unable to present this peptide by itself, augmented the T cell response at lower concentrations of peptide. These results suggest that the conserved alpha 3 domain of the class I MHC heavy chain as well as polymorphic regions play an important role in T cell activation and that T cell interaction with MHC molecules not presenting peptide can still augment the response. PMID- 8241056 TI - Influence of age on the signal transduction of T cells in mice. AB - A 5- to 10-fold decline was observed in the proliferative activity of T cells stimulated with anti-CD3 mAb between young and old mice. However, the number of CD3 molecules on the T cell surface was almost comparable between young and old T cells. The formation of the second messenger such as inositol triphosphate (IP3) and diacylglycerol (DAG) after mitogenic stimulation decreased in old T cells as compared with young ones. The activity of phospholipase C (PLC), which is responsible for the liberation of IP3 and DAG from phosphatidylinositol-4,5 bisphosphate (PIP2) was not different between young and old T cells. The content of PIP2 in the membrane was also comparable between young and old T cells. These findings have suggested that the age-related decline in the proliferative activity of T cells could be due to impairment of intracellular signal transduction, probably in the pathway somewhere between TCR and PLC. PMID- 8241057 TI - IL-5 receptor positive B cells, but not eosinophils, are functionally and numerically influenced in mice carrying the X-linked immune defect. AB - Mouse IL-5 (mIL-5) acts on B cells and eosinophils to induce growth and differentiation through the mIL-5 specific receptor (mIL-5R). The functional high affinity mIL-5R is a heterodimer composed of alpha and beta chains. We investigated the expression of mIL-5R and the responsiveness of B cells and eosinophils to mIL-5 in X-linked immunodeficient (xid) mice. mIL-5R expression analyzed by using mAbs specific for alpha and beta chains revealed that xid B cells had fewer mIL-5R alpha +mIL-5R beta + than BALB/c B cells. In particular, a decrease in the number of peritoneal mIL-5R+ B cells among Ly-1 B cells (known as B-1 cells) was remarkable. Furthermore, the frequency of precursors of mIL-5 responsive B cells in xid mice was approximately 100-fold lower than that of BALB/c mice. Interestingly, sorted mIL-5R+ peritoneal B cells from xid mice displayed a low response to mIL-5. Intraperitoneal injection of mIL-5 into BALB/c mice induced polyclonal IgM production and an increase in the number of eosinophils. The same regimen failed to induce an increase in the same parameters in xid mice. However, xid mice showed mIL-5-induced eosinophilia in peripheral blood to a similar extent as BALB/c mice. Eosinophils from mIL-5-injected xid mice expressed both alpha and beta chains of mIL-5, and responded to mIL-5 with prolonged in vitro survival. PMID- 8241058 TI - Adsorption behavior of fibrinogen to sulfonated polyethyleneoxide-grafted polyurethane surfaces. AB - Fibrinogen adsorptions to surface modified polyurethanes (PU, PU-PEO, and PU-PEO SO3) were studied from plasma in vitro. PU and PU-PEO surfaces demonstrated that initial adsorption increases with increasing plasma concentration in kinetic profiles and adsorption time in adsorption profiles as a function of plasma concentration, but after the plateau is reached, its adsorption amount decreases as plasma concentration (0.2-2.0%) and adsorption time (1-120 min) increase, respectively. In contrast, PU-PEO-SO3 showed that initial adsorption is almost same regardless of plasma concentration and adsorption time, which is due to the high affinity of surface sulfonate group to fibrinogen. All the surfaces indicated the Vroman effect at about 0.6% plasma concentration; however, the displacement was relatively low. Adsorbed amount of fibrinogen at steady state decreased in the order: PU > PU-PEO-SO3 > PU-PEO, regardless of adsorption time and plasma concentration. The adsorption behavior of PU-PEO-SO3 is attributed to both effect of low binding affinity of PEO chain and high affinity of pendant sulfonate group toward fibrinogen. PMID- 8241059 TI - Festschrift in honor of the 60th birthday of Dr. Allan S. Hoffman. Part 2. PMID- 8241060 TI - Basic fibroblast growth factor production in vitro by macrophages exposed to Dacron and polyglactin 910. AB - Macrophage activation by implanted blood-contacting biomaterials modulates smooth muscle cell and endothelial cell ingrowth. The present study evaluates the in vitro interactions between Dacron or polyglactin 910 with macrophages derived from rabbits fed either normal or atherogenic diets. Peritoneal macrophages were cultured in the presence or absence (negative controls) of either biomaterial for 7 weeks. Conditioned media was evaluated for mitogenic activity using a rabbit aortic smooth muscle cell bioassay with or without preincubation with neutralizing anti-basic-FGF antibody. Results demonstrated increased mitogen release from macrophages harvested from the atherosclerotic rabbits. Only macrophages harvested from normal diet fed rabbits increased their mitogen release following exposure to either polyglactin 910 (p < 0.05) or to Dacron (p < 0.005) over controls. The stimulation of mitogen release by polyglactin 910 did not significantly exceed that in response to Dacron. In rabbits fed normal diets neutralization with the anti-basic-FGF antibody inhibited 100% of the Dacron induced mitogen release as compared to 36% of the polyglactin 910 induced mitogen release (p < 0.01). These results demonstrate significant induced mitogen release from macrophages exposed to biomaterials in vitro, much of the smooth muscle cell mitogen represented by basic-FGF. PMID- 8241061 TI - Polyisobutylene-toughened poly(methyl methacrylate): III. PMMA-l-PIB networks as bone cements. AB - A series of novel polyisobutylene (PIB)-toughened poly(methyl methacrylate) (PMMA) networks consist of rubbery PIB domains covalently bonded to a glassy PMMA matrix. Materials containing 8.5-17 wt.% PIB (Mn = 18,000 g/mol) in a PMMA matrix (PIB/PMMA) were evaluated to assess their feasibility as the powder component along with methyl methacrylate (MMA) as the liquid component in a standard powder/liquid bone cement formulation. A standard ISO four-point bend test, commonly used for testing bone cements, was employed to investigate flexural properties. The mixing time and powder/liquid (P/L) ratio were studied to formulate novel PIB/PMMA cements for optimum toughness. Appropriate formulations led to improved toughness while adequate flexural strength and modulus were maintained. An experimental PIB/PMMA-system exhibited approximately 57 MPa flexural strength and approximately 2000 MPa flexural modulus. PMID- 8241062 TI - A model for thromboembolization on biomaterials. AB - A model was developed to describe the kinetics of protein and platelet deposition and embolization on biomaterials. The model assumes that proteins can be adequately represented by fibrinogen, albumin, and Factor XII, that protein adsorption is Langmuir-type, that surfaces are homogeneous, and that all adsorption and deposition steps are first order. Eleven model parameters were determined from literature experimental data from ex vivo experiments utilizing canine and baboon blood on Silastic, one parameter came from adsorption of Factor XII on glass, and three parameters were obtained by minimizing differences between experimental and predicted fibrinogen adsorption, and platelet deposition and embolization behavior. The model well predicted observed behavior for fibrinogen adsorption, platelet deposition, and platelet embolization on Silastic, and platelet embolization from both polyacrylamide and HEMA-MAAC. PMID- 8241063 TI - A wettability gradient as a tool to study protein adsorption and cell adhesion on polymer surfaces. AB - A new method for preparing a wettability gradient on polymer surfaces was developed. Low density polyethylene sheets were treated in air with corona from a knife-type electrode whose power gradually increases along the same length. The polymer surfaces oxidized gradually with the increasing power and the wettability gradient was created on the surfaces as evidenced by the measurement of water contact angles, Fourier-transform infrared spectroscopy in the attenuated total reflectance mode, and electron spectroscopy for chemical analysis. The wettability gradient surfaces prepared were used to investigate the interactions of model protein and cells in terms of the surface hydrophilicity/hydrophobicity of polymeric materials. PMID- 8241064 TI - Polyanionic hydrogel as a gastric retentive system. AB - Gastric emptying of a polyanionic hydrogel, polycarbophil (PC), from the canine stomach was studied using a duodenal cannulation technique. The basis of the study is to employ a certain quantity of swelling hydrogel that, by virtue of its swollen size and viscosity, converts a fasted stomach to a fed state and resists discharge from the stomach for an extended period of time. Different amounts of PC, in 200 ml water plus buffering agents, were administered orally to fasted canines. The gastric emptying lag time was found to increase with the viscosity of the administered dose. Addition of a base, sodium bicarbonate, to PC increased gastric retention via an increase in its apparent viscosity. The polymer mass is retained in the canine stomach until a sufficient quantity of stomach acid secretion reduces the viscosity of the viscous mass, through protonation of polycarbophil, and, at that point, discharge of the hydrogel will commence. Thus, gastric retention of PC hydrogel in the canine stomach can be prolonged by increasing the apparent viscosity of the hydrogel administered. PMID- 8241065 TI - Synthesis and characterization of a soluble, temperature-sensitive polymer conjugated enzyme. AB - The enzyme, alkaline phosphatase, has been conjugated to a temperature-sensitive polymer which exhibits a lower critical solution temperature (LCST). A series of copolymers containing different molar ratios of N-isopropylacrylamide(NIPAAm) and N-acryloxysuccinimide(NAS) were synthesized and then conjugated to the enzyme. These polymer-enzyme conjugates precipitate and flocculate in aqueous solution above the LCST, and redissolve when cooled below that temperature. The kinetics of the conjugated enzymes have been characterized as a function of temperature and compared to free enzyme. The effect of the conjugation degree between polymer and enzyme on the activity of the conjugated enzymes was also investigated. PMID- 8241066 TI - Synthesis and characterization of a new biodegradable semi-solid poly(ortho ester) for drug delivery systems. AB - Since the late 1970s, three families of poly(ortho esters) (POE) were synthesized to provide bioerodible carriers for drug delivery devices. The most recent POE is a semi-solid polymer with a viscous behavior at room temperature. Polymer synthesis by a transesterification reaction between a triol and a trialkyl ortho ester is described. The structure of the polymer was confirmed by conventional methods such as 1H-NMR, 13C-NMR and FT-IR. Information concerning average molecular weight and intrinsic viscosity was obtained respectively by GPC and viscosimetry. Residual solvents in the polymer were determined using gas chromatography. The chromatographic conditions were optimized to enable the quantification of the solvents in concentrations of a few percent. The mechanical behavior of the semi-solid POE was determined by rheometric measurements. Hydrolysis of the polymer leads to the formation of the original triol and the carboxylic acid derived from the trialkyl ortho ester used in the transesterification step. No toxicological problems associated with these compounds are anticipated. PMID- 8241067 TI - Investigations of the architecture of tamarind seed polysaccharide in aqueous solution by different scattering techniques. AB - The architecture of tamarind seed polysaccharide (TSP) has been investigated by light scattering (LS), small angle X-ray scattering (SAXS) and synchrotron radiation scattering (SRSAXS). The experimental data show that TSP in aqueous solution consists of multi stranded aggregates, with a high degree of particle stiffness. The angular dependence of the scattered intensity is typical for wormlike chains. Data evaluation on the basis of this model yields a statistical Kuhn segment length lK = 150 nm. The cross sectional radius of gyration is estimated as Rgcs = 6.0 +/- 0.5 A, which is more than twice the value, published for single stranded polysaccharides. Correspondingly, the experimental value of the linear mass density, measured by LS, is about five times higher than the theoretical value calculated from the primary structure. PMID- 8241068 TI - Synthesis and characterization of putrescine-based poly(phosphoester-urethanes). AB - A novel set of putrescine-based segmented polyurethanes was synthesized using 1,4 butane-diisocyanate and phosphoester diols, and was characterized for its potential as a degradable biomaterial. These poly(phosphoester-urethanes) (PPU) were flexible polymers with ultimate tensile strength (UTS) from 2 to 3 MPa, elongations up to 80% and tan delta near 0.15. The incorporation of phosphoester bonds in the backbone of the polymer by using bis(2-hydroxyethyl)phosphite (BGP) and bis(6-hydroxyhexyl)phosphite (BHP) as chain extenders resulted in hydrolytic degradation which was evaluated in vitro. By varying the content of the phosphoester diol BGP, degradation rate, as followed by mass loss and GPC, could be modulated. Polymers based on the more hydrophobic monomer, BHP, showed slower degradation than corresponding BGP based polymers. Tensile properties of PPU-B2 after 22 days in vitro degradation show more than a 50% drop in UTS and ultimate elongation, likely caused by void spaces left behind in the polymer after mass loss and swelling. The attachment of a drug, PAS, pendant to the phosphoester group of the PPU was demonstrated. PAS was linked via the spacer 4 hydroxybenzaldehyde, and free, intact drug was released in about 5 h from a thin film. PMID- 8241069 TI - Swelling controlled zero order and sigmoidal drug release from thermo-responsive poly(N-isopropylacrylamide-co-butyl methacrylate) hydrogel. AB - Thermo-responsive hydrogels of poly(N-isopropylacrylamide-co-butyl methacrylate) (poly-(IPAAm-co-BMA)) are capable of swelling-deswelling changes in response to external temperature. As poly(IPAAm-co-BMA) gels swell larger at a lower temperature, the degree and rate of the swelling could be controlled by temperature without altering the chemical structure. Therefore, drug release profiles were remarkably changed by alternation of temperature. The release profiles of indomethacin from poly(IPAAm-co-BMA) were observed to be zero-order at 20 degrees C. This release profile was explained in terms of a Case-II diffusion mechanism; which indicates relaxation of polymer chains with swelling was rate-determining. In the case of 10 degrees C, release demonstrated a sigmoidal profile. The acceleration of drug release was due to a rapid increase in swelling with disappearance of the glassy core which had constrained swelling. The regulation of the water-uptake process by changing external temperature remarkably affected drug release and resulted in several different release profiles. PMID- 8241070 TI - Chemical modification of biopolymers--mechanism of model graft copolymerization of chitosan. AB - Mechanism of graft copolymerization of vinyl monomers onto chitosan initiated by Ce(IV) ion, one of the important ways of chemical modification of chitosan, has been investigated by means of kinetics measurement and polymer chain structure analysis. It is found that when Ce(IV) ions reacts with adjacent hydroxylamine structure in chitosan, a chelate complex is first formed and then it disproportionates to radical for initiation of graft copolymerization. There exist two ways for such initiation depending upon the reaction temperature. For a reaction temperature lower than 40 degrees C, a -CHO group and a -C = NH group are introduced, and the aldehyde group reacts with Ce(IV) ions to form an acyl radical which initiates a graft polymer chain in this saccharide unit. For temperatures higher than 90 degrees C, the -C = NH group hydrolyzes to form an amine and an aldehyde group which also reacts with Ce(IV) ions. In this case there should be two aldehyde groups, i.e. two initiation sites, in one adjacent hydroxyl-amine structure. That means that the initial radical in the chitosan/Ce(IV) system is similar to that in the cellulose/Ce(IV) one, but the former usually gives a lower grafting reactivity than the latter due to the higher stability of the chelate. PMID- 8241071 TI - Production of staphylococcal impetigo-like lesion on human skin explants in culture. AB - We produced a highly reproducible experimental impetigo-like lesion in normal human skin explants in culture. The three Staphylococcus aureus strains we used were an isolate from a human impetigo (E strain), an isolate from a human furunculosis (N strain) and ATCC 29213 strain. E strain was a protein A positive, coagulase type V, producer of exfoliative toxin (ET) and beta-toxin. N strain was a coagulase type IV, ET non-producer and alpha-toxin positive. ATCC 29213 was a coagulase type II, ET non-producer, and alpha-, beta-, and delta-toxin positive. Normal human skin samples were obtained from 8 adult skin surgery patients. One specimen was obtained from human oral mucosa. Small pieces of the samples were slightly abraded on the epidermal surface and cultured on lens paper rafts floating in Eagle's Minimum Essential Medium in an atmosphere of 5% CO2 and 95% air. Fifty microliters of the respective bacterial suspensions were applied to the epidermal surfaces of the explants. The inoculated surfaces were then occluded under sterile plastic plaster. Histologically, the formation of intraepidermal blisters at the granular layer level with acantholytic cells was observed in all 8 of the skin specimens at 10 h after inoculation with E strain. The specimen from an oral mucous membrane did not produce similar changes with any of the three S. aureus strains. Neither N or ATCC strains developed bullae in the epidermis at 6, 10 or 18 h after inoculation. Immunofluorescent examination revealed that the inner surfaces of blisters in the epidermis were lined with anti-ETA antibody. Under the electron microscope, the blisters of the specimens which had been inoculated with strain E contained only a few S. aureus cells. These results suggest that blister formation at the granular layer level with acantholytic cells is mediated by ET action at the granular layer level and occurs without invasion of lymphocytes or neutrophils, or the involvement of any serum components. Therefore, under appropriate conditions, impetigo could develop even in adults. PMID- 8241072 TI - A cross-reaction between piroxicam-photosensitivity and thiosalicylate hypersensitivity in lymphocyte proliferation test. AB - We examined the cross-reaction between photosensitivity to piroxicam (PXM) and contact sensitivity to thiosalicylate (TOS) by a lymphocyte proliferation test (LPT) in guinea pigs. The lymph node cells (LNCs) plus peritoneal exudate cells (PECs) from guinea pigs contact-sensitized with TOS remarkably cross-proliferated to PXM under UVA (4 J/cm2) irradiation. On the other hand, the PXM photosensitized LNCs+PECs also cross-proliferated to TOS. From these results, the reciprocal cross-reaction between TOS-hypersensitivity and PXM-photosensitivity was reconfirmed by the in vitro LPT, indirectly indicating that the PXM photosensitivity is a cell (probably T cell)-mediated PXM photoallergy in its nature. The TOS-primed LNCs+PECs did not cross-proliferate to UVA (4 J, 180 J or 500 J/cm2)-pretreated PXM (UVA-PXM) although it is supposed to contain several photoproducts of PXM. Furthermore, the TOS-primed LNCs developed a remarkable proliferative cross-response to the PECs pulsed with PXM under UVA (4 J/cm2) irradiation (photo-PXM-modified PECs), but not to the PECs pulsed with PXM or UVA PXM. Therefore, it is presumed that the cross-reactive molecule, which is easily formed from PXM under UVA irradiation, is unstable, and that the formation of complete antigen by the generation of this molecule and its photobinding needs the coexistence of PECs, PXM and UVA irradiation at the same time in the culture. PMID- 8241073 TI - The effect of aging and arteriosclerosis on human skin blood flow. AB - Skin blood flow in the deltoid region of 65 men was measured by the xenon 133 clearance method and in studying the correlation of age, total cholesterol and systolic blood pressure to skin blood flow, the following results were obtained. (1) Skin blood flow significantly decreased with increase in age. In calculating skin blood flow at 20 and 70 years of age from the regression line between skin blood flow and age, it was estimated that skin blood flow at 70 years of age decreased to 40% of that at 20 years of age. (2) Skin blood flow significantly decreased with increase in total cholesterol and systolic blood pressure. (3) Skin blood flow showed the closest correlation with age, followed by total cholesterol and systolic blood pressure which showed a weak correlation. It is estimated from the foregoing results that as skin blood flow in patients with arteriosclerosis decreases more in elderly individuals than in young persons, their wound healing is delayed. PMID- 8241074 TI - A speculative view of atopic dermatitis: barrier dysfunction in pathogenesis. AB - The development of atopy or atopic dermatitis is undoubtedly influenced by immunological abnormality. However, the question of factors which induce tolerance breakthrough and hypersensitivity to non-selective multiple antigens in atopy has remained unanswered. The broad clinical spectrum of this disease cannot be explained simply by allergy or immunological abnormalities. This speculative article is the first attempt to answer these questions and to explain the concept of atopic dermatitis by the barrier dysfunction theory based on our experimental results which demonstrated and increased permeability of the stratum corneum in patients with atopic dermatitis. We consider immunological abnormality and mucocutaneous barrier dysfunction as two major defects of atopy or atopic dermatitis. The wide range of clinical manifestations presented by this disease can be more easily comprehended by partial overlapping of these two core abnormalities. A mucocutaneous barrier defect readily allows penetration of multiple antigens or haptens. Repeated encounters with allergens may induce not only tolerance breakthrough and non-selective hypersensitivities but also enhancement of the allergic inflammation. On the other hand, an allergic inflammation stemming from the immunological abnormalities breaks down barrier functions. This sequence results in a 'vicious circle', which plays the most important role in pathogenesis of atopic dermatitis and probably of other atopic diseases. PMID- 8241076 TI - Abstracts from the 2nd Tricontinental Meeting of JSID, SID and ESRD. Kyoto, Japan, October 28-31, 1993. PMID- 8241075 TI - Successful management of Hailey-Hailey disease with potent topical steroid ointment. AB - In order to know the real effect of steroid for Hailey-Hailey disease, seven patients were treated with strong steroid ointments in conjunction with antibiotics and/or antimycotic ointments. All patients were infected by various kind of bacteria and fungus. All patients showed remission within 2-16 weeks. Four patients maintained remission from 1.5 to over 4 years when ointments were continuously applied. No serious side-effects except skin atrophy and contact dermatitis were observed. These findings suggest that strong steroid ointment was really effective for this disease when secondary infection was prevented by the application of antibiotics and/or antimycotics, and that this disease is controllable with strong steroid ointments. PMID- 8241077 TI - [Transmission electron microscopic ultrastructures of the male germinal cells of Fibricola seoulensis]. AB - A transmission electron microscopic study was performed to observe the ultrastructures of the male germinal cells and spermatozoa of Fibricola seoulensis. Spermatogonia were found in the periphery of the testis and characterized by large nuclei and comparatively little cytoplasms. Spermatocytes contained an oval to spherical nucleus. Their nuclear volume was little larger in comparative to that of cytoplasm, and the chromatin was comparatively little. The early spermatids were characterized by a great amount of cytoplasm, and numerous mitochondria encircled the nucleus. In a more advanced spermatids the electron dense strands of chromatin appeared in the nucleus, and a pair of rootlet of the axoneme and a microtubule-organizing center (MTOC) were observed near the nucleus. The sectioned spermatozoa were found in the testis and the seminal vesicle. Their cross sectional views were divided into 6 types when they were distinguished on the basis of the morphology and components. The spermatozoa of F. seoulensis showed two flagella of 9+1 type axoneme. PMID- 8241078 TI - Studies on isosporosis in dogs. I: Isolation and sporulation of Isospora ohioensis. AB - Based on the morphologic characteristics of oocysts of the genus Isospora, we have demonstrated that I. ohioensis is a relatively common Isospora species in dogs which are resident in Chonbuk province, Korea. The prepatent period of I. ohioensis was four days. The size of the unsporulated oocysts in fresh stool specimens was 22.9 x 19.8 microns (R = 1.16). The size of the contracted sporonts was 17.4 x 16.3 microns (R = 1.06). By 96 hours, sporulation is complete and the ratio of length/width was constant relatively. And the sizes of oocysts and sporocysts were 22.8 x 20.5 microns (R = 1.11) and 15.0 x 10.8 microns (R = 1.39), respectively. At this time it is most reliable for the measurements of oocyst sizes of I. ohioensis to provide with the identifiable clues against the others. It is therefore recommended that the clinical fecal specimens suspected of isosporosis should first be incubated and aerated for 96 hours before a definitive parasitological diagnosis can be reached. These and other observations contribute to the understanding of the biological characteristics and laboratory and clinical diagnosis of isosporosis. PMID- 8241079 TI - [Current status and the changing pattern of the prevalence of clonorchiasis in the inhabitants in Sanchong-gun, Kyongsangnam-do, Korea]. AB - This study was conducted to evaluate the current endemicity and the transition mode of the Clonorchis infection in Sanchong-gun Kyongsangnam-do. The areas of investigation, villages and schools surveyed, method and techniques used in this study were the same as in 1984. The egg positive rate of clonorchiasis in the general population of Sanchong-gun was 37.6% out of 837 examined persons. In the schoolchildren, the rate was 9.7% out of 145 persons examined. In the intensity of the infection among the cases, the mean EPG (Egg per gram of feces) in the inhabitants and schoolchildren were 3,310 (male 4,221, female 1,978) and 711 (male 620, female 833), respectively. Of the inhabitants with clonorchiasis, the cases of light infection (EPG < 1,000) and moderate infection (1,000 < EPG < 10,000) were 93.9%. The proportion with experience of eating raw freshwater-fish was 90.5% among the infecteds. Out of 138 inhabitants examined in both 1984 and 1992, the number of the positives by both examinations was 31 (22.5%), positive to negative conversion was 35 (25.4%), negative to positive conversion was 22 (15.9%). The present results reveal that there are no changes of the egg positive rate of clonorchiasis among inhabitants in Sanchong and the eating habit of raw freshwater-fish persists there. In this community, the eating habit is the most important problem to be solved for control of this endemic disease. PMID- 8241080 TI - [Comparative susceptibility of different cell lines for culture of Toxoplasma gondii in vitro]. AB - In order to establish a useful cell culture system for T. gondii, we compared the degree of proliferation of T. gondii tachyzoites among 8 different cell lines; 2 kinds of normal animal cells (MDCK-canine kidney cells; Vero-monkey kidney cells) and 6 kinds of human tumor cells (A 549, PC 14-lung cancer cells; SNU 1, SNU 16, MKN 45-stomach cancer cells; HL-60-promyelocytic leukemia cells), through morphological observation and 3H-uracil uptake assay. The degree of susceptibility to infection with T. gondii tachyzoites was highest in A 549 and PC 14 cells, medium in Vero, HL-60, MDCK and SNU 1, and lowest in SNU 16 and MKN 45 cells. The kinetics of T. gondii multiplication during the post-infection 60 hours were highly dependent upon the dose of tachyzoites administered and the duration of cultivation. These results show that A 549 and PC 14 are the most suitable cell lines among the 8 tested for the growth and multiplication of T. gondii in vitro. PMID- 8241081 TI - Passive transfer of immunity against Cryptosporidium infection in neonatal mice using monoclonal antibodies. AB - Monoclonal antibodies (mAb) against merozoites and sporozoites of the protozoan parasite Cryptosporidium parvum were examined for potential modulation of cryptosporidial infections in vivo by daily oral mAb administration to oocyst inoculated neonatal mice. Monoclonal-treated neonatal mice were sacrificed four and eight days post infection (pi). Differences in infection rates were observed among the treatment groups at the p < 0.05 level. Suckling mice treated daily with orally administered mixtures of mAbs (ascitic fluids) showed significantly reduced parasite loads compared to control mice at four and eight days pi, while suckling mice receiving mAb Cmg-3 alone showed significant differences only at 4 days pi., suggesting that passive transfer of mAb may be of value in controlling cryptosporidial infections. PMID- 8241082 TI - Changes of IgE production, splenic helper and suppressor T lymphocytes in mice infected with Paragonimus westermani. AB - Effects of Paragonims westermani infection were observed in mice on the change of serum IgE level, the number of peripheral eosinophils and the distribution of Thy 1.2+ (CD3), L3T4+ (CD4), and Lyt-2+ (CD8) splenic T lymphocytes without mitogen stimulation. BALB/c mice were infected with 20 metacercariae of P. westermani each. Total serum IgE increased at 3 weeks after the infection and reached a peak on week 4 and maintained high levels of IgE until the 23rd week. Peripheral eosinophil numbers increased at the second week and attained peak level on week 9. The frequency of L3T4+ (CD4) and Lyt-2+ (CD8) T lymphocytes decreased slightly until 4 weeks after the infection, but not significantly. Absolute number of L3T4+ and Lyt-2+ T lymphocytes, and the ratio of L3T4/Lyt-2 were not markedly changed over the period of observation. The frequency of Thy 1.2+ (CD3) T lymphocytes in the infected group slightly decreased until 4 weeks after the infection and showed significant reductions at the 2nd and 4th week of the infection (p < 0.05). PMID- 8241083 TI - [Natural killer cell activity in mice infected with free-living amoeba with reference to their pathogenicity]. AB - The purpose of this observation was to investigate the natural killer cell activities in mice infected with pathogenic free-living amoeba, Naegleria fowleri and Acanthamoeba culbertsoni according to the infection doses, and infected with non-pathogenic free-living amoeba, Naegleria gruberi. The natural killer cell activity was examined by means of target binding capacity, active NK cell and maximum recycling capacity of the mice after inoculating free-living amoebae with low and high doses. The mice infected with 1,103, 1,105 A. culbertsoni trophozoites showed mortality rates of 6.9% and 65.5%, respectively. The mice infected with 1,104, 1,105 N. fowleri trophozoites showed mortality rates of 5.9% and 72.2%, respectively. The NK cell activities in all experimental groups increased significantly on day 1 after infection as compared with control group, and then remarkably declined thereafter, there was no difference of the cytotoxic activity of the NK cells in mice among inoculation doses of pathogenic free living amoebae. The target binding capacities of NK cells and percentages of activated NK cells in mice infected with pathogenic free-living amoebae were significantly increased a day after infection, as compared with control group. There was no difference of the maximal recycling capacities of NK cells in all experimental groups as compared with control group. There was significant difference in the cytotoxic activity and single cell cytotoxicity of NK cells between the experimental groups infected with pathogenic free-living amoebae and that infected with non-pathogenic free-living amoebae. PMID- 8241084 TI - [Interleukin-2 production and alteration of T cell subsets in mice infected with Naegleria fowleri]. AB - Naegleria fowleri is the cause of primary amoebic meningoencephalitis in man. IL 2 levels after stimulation of T lymphocytes by PHA or N. fowleri lysates, the amounts of T lymphocyte subsets and the blastogenic responses of T lymphocytes in mice after infected with pathogenic N. fowleri were studied comparing between two study groups, one 1 x 10(4) trophozoites inoculated mice and the other 1 x 10(5) trophozoites inoculated mice. All experimental samples were obtained on the day 7, 14 and 24 after inoculation. The mice inoculated with 1 x 10(4) trophozoites showed a 14.3% mortality rate, and 72.2% in the mice inoculated with 1 x 10(5) trophozoites. The IL-2 levels on day 14 of two experimental groups were significantly decreased as compared with the control group. Thy 1.2+ T cells in the total spleen lymphocytes of 1 x 10(5) trophozoites inoculated group on day 7 were significantly increased compared with the control group. There was no significant difference between 1 x 10(4) trophozoites inoculated group and the control group. L3T4+ T cells and Ly2+ T cells in the total spleen lymphocytes of 1 x 105 trophozoites inoculated group on day 7 were significantly increased compared with the control group. The DNA S fraction of T cells in the spleen of 1 x 10(5) trophozoites inoculated group was significantly increased on day 7. The amount of S fractions of DNA were sequentially decreased on day 14 and 24, but they were also significantly increased compared with the control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241085 TI - Characterization of a peroxidase in excretory-secretory product of adult Paragonimus westermani. AB - When activity of peroxidase in adult Paragonimus westermani was monitored using o dianisidine and H2O2 as substrates, its specific activity was 1.5 times higher in excretory-secretory product (ESP) than in crude extract. The enzyme was purified by two purification steps of Sephacryl S-300 Superfine gel permeation and DEAE Trisacryl M anion exchange chromatographies. Its activity increased 16.9 fold with 32.3% recovery. The enzyme was inhibited totally by 1 millimoles of dithiothreitol (DTT), 2-mercaptoethanol and azide. Molecular mass was 16 kDa in reducing SDS-polyacrylamide gel electrophoresis (PAGE) or 19 kDa in TSK-Blue gel filtration high performance liquid chromatography (HPLC), respectively. Special staining for peroxidase by diaminobenzidine on SDS-PAGE confirmed the activity. The peroxidase was less reactive to a paragonimiasis serum when observed by SDS PAGE/immunoblot. In addition, specific activities of superoxide dismutase (SOD) and catalase were also identified in the ESP. High activities of these antioxidant enzymes in ESP indicate that they are parts of defense mechanisms against reactive oxygen intermediates from host. PMID- 8241086 TI - Protein composition and antigenicity of the tegument from Paragonimus westermani. AB - To ascertain that tegument of Paragonimus westermani has specific antigenic proteins, the tegumental fraction was isolated from 10-month-old worms by 0.1% digitonin solution, and subjected to SDS-PAGE and immunoblot. Component proteins of tegumental syncytium comprised of 94, 74 (76-66), 62, 54, 44, 42, 38, 28, 26, 25, 24, 17, 15.5 and 13. 5 kDa proteins. Of them, the 94, 44 and 42 kDa proteins were more specific to tegument, especially the 94 kDa protein was the most prevailing one. In immunoblot, antigens of the 94, 90, 78, 76, 74, 68, 65, 62, 60, 59 and 54 kDa proteins were commonly detected by 7 sera of 10 human paragonimiasis, but none of them reacted with 5 sera of clonorchiasis. In conclusion, the 94 kDa protein was the major tegumental protein, as well as the specific antigen. The 76 and 66 kDa proteins were the minor components of tegument, which were also specific antigens of P. westermani. PMID- 8241087 TI - Influence of heat shock, drugs, and radiation on karyotype of Leishmania major. AB - Leishmaniasis is one of the important tropical diseases in the world. Although it is not prevalent in Korea, imported cases have been recorded. The karyotype of Leishmania sp. has been observed to be variable by localities or by strains, but the karyotype of a strain is known to be stable. This study was performed to observe if the karyotype of a Leishmania sp. would be changed under some stressful conditions. The karyotype, analyzed by pulsed field gradient gel electrophoresis, was not grossly changed by heat shock, chemotherapeutics, UV illumination, and gamma irradiation. Radiation destroyed the chromosomes mechanically, but subcultured organisms after irradiation showed unaffected karyotype. The present findings suggest that the karyotype of a Leishmania strain is so stable that it is not altered by temporary stimulation with heat, drugs, and radiation. PMID- 8241088 TI - Clonorchis sinensis tropomyosin: cloning and sequence of partial cDNA amplified by PCR. AB - C. sinensis total RNA was containing large amount of 18S rRNA but little 28S rRNA. The size of the double-stranded cDNA synthesized from poly (A)+ mRNA was 0.4-4.2 kb long with tapering upto 9.5 kb. Degenerated oligonucleotides (as 2 sense and 3 antisense primers) were designed on the conserved regions of the known tropomyosin amino acid sequences. From one out of the PCR amplifications using total cDNA and matrix of primers, a specific gene product, 580 bp in size, was produced. Upon Southern hybridization of the PCR products with Schistosoma mansoni tropomyosin (SMTM) cDNA, only one signal appeared at the band of 580 bp product. This 580 bp product was considered to encode C. sinensis tropomyosin (CSTM) and cloned in pGEM-3Zf(-) for DNA sequencing. CSTM cDNA was 575 bp containing one open reading frame of 191 predicted amino acids, which revealed 86.3% homology with SMTM and 51.1% with Trichostrongylus colubriformis tropomyosin. CSTM cDNA obtained will serve as a probe in the studies of molecular cloning of CSTM. PMID- 8241089 TI - Intestinal parasitic infections of Korean Army soldiers in Whachon-gun, Korea. AB - We examined stools of 113 soldiers of Korean Army in Whachon-gun, Korea in 1992. The eggs of Clonorchis sinensis (7/113) and Metagonimus sp. (3/113), and cysts of Giardia lamblia (4/113) were detected. No eggs of soil-transmitted helminths were found. Most of the Clonorchis-infected soldiers (6/7) were recruited from Kyongsangnam-do. PMID- 8241090 TI - [A case of pelvic paragonimiasis combined with myoma uteri and pelvic inflammatory disease]. AB - Paragonimus westermani is a lung fluke of humans that is usually found in the lungs but may be found elsewhere in many unusual locations. A case of pelvic paragonimiasis was found incidentally by surgical intervention of inflammatory disease and myoma uteri. She was a 51-year-old Korean woman complaining of lower abdominal pain and intermittent vaginal spotting. Numerous Paragonimus ova were observed in the resected omentum in the pelvis after total abdominal hysterectomy. It is suggested that pelvic paragonimiasis may be one of causative agents of pelvic inflammatory disease. PMID- 8241091 TI - Benzo(a)pyrene enhances atherosclerosis in White Carneau and Show Racer pigeons. AB - Benzo(a)pyrene (BaP), a major environmental pollutant and component of cigarette smoke, is both carcinogenic and atherogenic in experimental models. We investigated the effect of long-term administration of BaP on atherogenesis in both atherosclerosis-susceptible White Carneau (WC) and atherosclerosis-resistant Show Racer (SR) pigeons. The number and size of arterial lesions in the brachiocephalic arteries in WC and SR females but not males were significantly enhanced after long-term dosing with BaP. Metabolic activation appears to be required for BaP atherogenicity, since benzo(e)pyrene (BeP), a noncarcinogenic analogue of BaP, did not enhance lesion development. Studies with 3H-BaP revealed no significant differences between male and female or between WC and SR pigeons in the arterial distribution of BaP and/or its metabolites. There were no consistent differences in blood pressure or plasma cholesterol levels between breeds or sexes. However, chronic administration of BaP did result in complete infertility in female birds, concomitant with grossly visible changes in ovarian appearance. These results clearly show that long-term dosing with BaP alters ovarian structure and function in treated birds, at the same time aggravating the development of arterial lesions. Thus, BaP-induced atherogenicity in female pigeons may be a consequence of an alteration in estrogen production or of antiestrogenic properties of BaP at the level of the arterial wall and may serve as a highly useful animal model to examine the well-known rapid development of atherosclerosis in postmenopausal women. PMID- 8241092 TI - Familial hypoalphalipoproteinemia in premature coronary artery disease. AB - Hypoalphalipoproteinemia (HA) is a common finding in patients with premature coronary artery disease. To characterize the common familial forms of HA, we studied 102 families of probands with premature coronary artery disease; 40 probands (39.2%) had HA. Of these, 25 had at least one first-degree relative affected with HA; 11 had familial hypertriglyceridemia with HA (FTgHA); 10 had familial combined hyperlipidemia (FCH); and 4 had familial HA (FHA) with no other lipoprotein abnormalities. In the remaining 15 families, no lipoprotein abnormalities were observed in first-degree relatives. We measured apolipoprotein (apo) A-I, B, C-III, and E levels as well as lipoprotein particle (Lp) levels of LpA-I (containing apoA-I only), LpA-I:A-II (containing both apoA-I and A-II), LpB:E, and LpB:C-III. Compared with a reference group of healthy men (n = 103) and women (n = 106), probands with familial forms of HA had lower high-density lipoprotein cholesterol levels by selection criteria. Triglyceride levels were higher in FTgHA and FCH probands than in the reference group or FHA subjects. Despite selection of FTgHA and FCH by low-density lipoprotein (LDL) cholesterol, the latter was not significantly different between the three groups and the reference group. ApoA-I levels were decreased in FCH, FHA, and FTgHA probands, and LpA-I and LpA-I:A-II were lower in FHA and FTgHA probands. ApoB levels were significantly higher in all familial HA groups compared with the reference group, being highest in FCH individuals, but not significantly higher between FCH, FTgHA, or FHA probands. LpB:E levels were higher in the FCH and FTgHA groups than in the reference group. There were no significant differences between groups for apoE, apoC-III, and LpB:C-III. LDL particle size was smaller in all three forms of FHA, which, in combination with higher apoB levels, reflects an increased number of smaller, denser LDL particles. Affected children had, on average, higher apoB and LpB:E levels than nonaffected siblings. Our data suggest that common forms of FHA in subjects with coronary artery disease represent a spectrum of overlapping disorders characterized by an increase in apoB-containing lipoproteins, especially LpB:E particles, and smaller, denser LDL particles. When using appropriate age- and gender-adjusted cutpoints, approximately half the offspring (in young adulthood) appeared to be affected. PMID- 8241093 TI - Severity of peripheral atherosclerosis is associated with fibrinogen and degradation of cross-linked fibrin. AB - Immunohistochemical studies of human atherosclerotic lesions have demonstrated the occurrence of fibrin deposition and its degradation in the arterial wall. We studied fibrinogen, the generation of thrombin, and the degradation of fibrin in 40 patients with stable peripheral arterial occlusive disease of varying severity, as assessed by the ankle/brachial pressure index and duplex ultrasonography and/or angiography. Circulating fibrinogen (functional and immunological), fibrinopeptide A, thrombin-antithrombin III complex, and D-dimer were measured. The severity of atherosclerosis was associated with both fibrinogen (both functional and immunological) and D-dimer (r = .57, P < .0002, and r = .57, P < .0001, respectively). Fibrinogen and D-dimer showed a significant positive correlation (r = .50, P < .001). Generation of thrombin was detected in 24 patients (60%) by fibrinopeptide A and levels of thrombin antithrombin III complex. As a sign of coagulation activation and fibrinolysis, we found that thrombin-antithrombin III complex and the degradation of cross linked fibrin were progressively associated with the extent of vascular disease. The plasmin-mediated fibrin breakdown contributed to increased levels of circulating fibrinogen, an established risk factor for thrombotic complications. The significant correlations between fibrinogen/D-dimer and the severity of atherosclerosis support previous pathological studies and imply that local degradation of cross-linked fibrin is involved in the progression of atherosclerosis. PMID- 8241094 TI - Influence of triacylglycerol biosynthesis rate on the assembly of apoB-100 containing lipoproteins in Hep G2 cells. AB - Apolipoprotein B-100 (apoB-100) appears in three forms in the endoplasmic reticulum of Hep G2 cells: (1) tightly bound to the membrane, ie, not extractable by sodium carbonate. This form is glycosylated but protease sensitive when present in intact microsomes, suggesting that it is only partially translocated to the microsomal lumen; (2) extractable by sodium carbonate and present on low density lipoprotein-very-low-density lipoprotein (LDL-VLDL)-like particles. This form is glycosylated and secreted into the medium; and (3) extractable by sodium carbonate but having a higher density than the LDL-VLDL-like particles. This form, referred to as Fraction I, is glycosylated and protected against proteases when present in intact microsomal vesicles, indicating that it is completely translocated to the luminal side of the microsomal membrane. Fraction I is not secreted into the medium, but it disappears with time from the cell, suggesting that it is degraded. Oleic acid induced a 2.7-fold increase in the rate of the biosynthesis of triacylglycerol but not of phosphatidylcholine in Hep G2 cells. Incubation of the cells with oleic acid had no significant effect on the rate of initiation of the apoB-100-containing lipoproteins, nor did it influence the amount of apoB-100 that was associated with the membrane or the turnover of apoB 100 in the membrane. Instead, it increased the proportion of the nascent apoB polypeptides on initiated lipoproteins that was converted into full-length apoB 100 on LDL-VLDL-like particles, giving rise to an increased amount of these particles in the lumen of the secretory pathway. Pulse-chase experiments showed that incubation with oleic acid gave rise to an increased formation of LDL-VLDL like particles on behalf of the formation of Fraction I. This effect of oleic acid could partially explain the protective effect of the fatty acid on apoB-100, preventing it from undergoing posttranslational degradation. PMID- 8241095 TI - Effect of pravastatin and omega-3 fatty acids on plasma lipids and lipoproteins in patients with combined hyperlipidemia. AB - This study compared the effects of a 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor, fish oil, and placebo on plasma lipids and lipoproteins in patients with mixed hyperlipidemia. After an initial run-in phase, 32 patients were randomized for 6 weeks to either (1) pravastatin 40 mg/d, n = 10; (2) fish oil (himega 6 g/d, equivalent to 3 g omega-3 fatty acids/d), n = 10; or (3) placebo. After single drug therapy, in the pravastatin group mean total plasma cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and apolipoprotein (apo) B fell significantly by 23% (P < .001), 30% (p < .001), and 26% (P < .01), respectively. LDL Stokes' diameter did not change. In the fish oil group mean plasma triglycerides (TG) fell 30% (P < .05), LDL Stokes' diameter increased from 25.0 to 25.9 nm (P < .05), and there was a nonsignificant increase in LDL-C. There were no changes in the placebo group. To assess the effect of the combination of pravastatin plus fish oil therapy, all patients, except one woman from the placebo group who developed nausea on fish oil, then took combined therapy of pravastatin 40 mg/d plus fish oil 6 g/d for an additional 12 weeks. In each case, there were no clinically significant episodes of muscle tenderness or elevation of creatine phosphokinase or alanine aminotransferase. After 12 weeks of combined therapy of pravastatin plus fish oil, there were significant reductions in the mean TC, TG, LDL-C, and apoB in the three groups compared with baseline levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241096 TI - Staining of endothelial cells and macrophages in atherosclerotic lesions with human heat-shock protein-reactive antisera. AB - Our previous epidemiological studies have shown that levels of serum antibodies against mycobacterial heat-shock protein (hsp) 65 correlate positively with carotid atherosclerosis in subjects aged 40 to 79 years. To determine whether these high-titer sera also react with homologous human hsp60 and/or cell components of atherosclerotic lesions, we selected 15 human sera samples, each with high or low titers to recombinant mycobacterial hsp65, and investigated their reactivity with human arterial lesion components by immunoblotting and immunofluorescence techniques. All five higher-titer sera against hsp65 reacted with a 60-kDa band of atherosclerotic lesion proteins and human recombinant hsp60 on Western blots. Pooled sera with low antibody titers to hsp65 diluted similarly as high-titer sera did not show reactivity with atherosclerotic lesion and media proteins. By immunohistochemistry and immunofluorescence with human immunoglobulin G isolated from different sera, labeled with biotin, and visualized with a streptavidin conjugate, positive staining was observed in sections of fatty streaks and atherosclerotic plaques of carotid arteries, and weak staining was observed in the normal intima. Double immunofluorescence identified the majority of positively stained cells as macrophages, endothelial cells, and a few smooth muscle cells. In summary, serum antibodies against hsp65 cross-react with the human 60-kDa homologue present in high levels in atherosclerotic lesions and are mainly reacting with macrophages and endothelial cells, supporting our concept of a possible involvement of humoral-mediated immune reaction against hsp60 in atherogenesis. PMID- 8241097 TI - Comparative study of the activity and composition of HDL3 in Russian and American men. AB - Previous studies conducted within the framework of the Lipid Research Clinics Program showed a strong inverse correlation between high-density lipoprotein cholesterol (HDL-C) level and coronary heart disease (CHD) risk in American male populations, whereas in Russian populations such a correlation was less pronounced. It was assumed that HDL was less protective of CHD in Russian than in American males. This study compared the functional activity and lipid composition of HDL3 isolated from the blood plasma of men with low, normal, and high HDL-C levels from Moscow (Russia) and Seattle (United States) populations. Results obtained showed that American HDL3, irrespective of the plasma HDL-C level, had higher activity in stimulating both [3H]cholesterol and cholesterol mass efflux from cholesterol-loaded fibroblasts and in suppressing cellular cholesterol esterification when compared with Russian HDL3. American HDL3 remained more active than Russian HDL3, even when apolipoprotein E-containing particles were removed from HDL3 by heparin-Sepharose affinity chromatography. Russian and American 125I-HDL3 had similar binding to high-affinity cell-surface sites, but Russian HDL3 had a higher nonspecific binding component compared with American HDL3. This study demonstrates for the first time potential functional differences between HDL particles isolated from Russian and American populations. The lower activity of Russian HDL3 in promoting cellular cholesterol efflux may partly explain the higher CHD risk in the Russian population compared with the American one. PMID- 8241098 TI - Vitamin E, LDL, and endothelium. Brief oral vitamin supplementation prevents oxidized LDL-mediated vascular injury in vitro. AB - In previously reported in vitro studies, we found that heme, a physiologically widespread hydrophobic iron compound, can rapidly generate oxidized low-density lipoprotein (LDL), which then becomes cytotoxic to cultured vascular endothelial cells; both LDL oxidation and endothelial cytotoxicity were inhibited by incubation with exogenous alpha-tocopherol (vitamin E) or ascorbic acid (vitamin C). Seeking relevance to in vivo conditions, we performed a study in which 10 human volunteers were given daily antioxidant supplements of 800 IU of DL-alpha tocopherol acetate alone or in combination with 1000 mg of ascorbic acid for 2 weeks. LDL resistance to heme oxidation ex vivo, as measured by the lag time for conjugated-diene formation, increased by as much as threefold from a mean +/- SD of 58 +/- 11 to 104 +/- 18 minutes (P < .001); LDL alpha-tocopherol increased from 11 +/- 2 to 26 +/- 6 molecules per LDL particle (P < .001); and most impressively, cytotoxicity to porcine aortic endothelial cells incubated with LDL conditioned with heme plus H2O2 or with copper was completely prevented (cytotoxicity before supplementation was 42 +/- 12%, decreasing after supplementation to 3 +/- 2%, P < .001). These measurements reverted to their presupplement levels within 2 weeks after participants stopped taking antioxidant supplements and were reproduced in 4 subjects taking 800 IU of DL-alpha tocopherol acetate supplements alone but not in the same subjects taking 1000 mg ascorbic acid supplements alone. In conclusion, oral vitamin E supplementation increases LDL alpha-tocopherol content, increases LDL resistance to oxidation, and decreases the cytotoxicity of oxidized LDL to cultured vascular endothelial cells. PMID- 8241099 TI - Influence of probucol on enhanced LDL oxidation after fish oil treatment of hypertriglyceridemic patients. AB - The susceptibility of low-density lipoprotein (LDL) to oxidation was studied in hypertriglyceridemic men (5 with type III and 5 with type IV) at baseline on a low-saturated-fat, low-cholesterol diet, after 6 weeks of dietary supplementation with fish oil (Promega, 12 g/d), and after 6 weeks of fish oil combined with probucol (500 mg BID). The relative content of n-3 polyunsaturated fatty acids in plasma and LDL was increased during the two treatment periods, and a low alpha tocopherol to n-3 polyunsaturated fatty acids ratio was observed. Plasma thiobarbituric acid-reactive substances (TBARS) levels were unchanged after 6 weeks of fish oil, but the ratio of lipid peroxides to the reduced triglyceride (TG) levels (MDA:TG) was significantly higher (P < .01). Addition of probucol lowered both absolute levels of TBARS (P < .01) and the MDA to TG ratio (P < .001). The susceptibility of LDL to Cu(2+)-catalyzed oxidation was evaluated over a 5-hour time course by determining TBARS formation, free amino group levels, and changes in LDL electrophoretic mobility. TBARS levels that were higher in native LDL (1.019 < d < 1.050 g/mL) after 6 weeks of fish oil than at baseline (P < .01) were reduced 52.3 +/- 11.3% by the addition of probucol (P < .001). With fish oil alone, TBARS production after exposure of LDL to Cu2+ for 5 hours was increased 17.0 +/- 5.8% compared with corresponding baseline values (P < .001), whereas a 64.1 +/- 14.3% reduction from the previous period was observed with fish oil + probucol (P < .001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241100 TI - Identification of novel heparin-releasable proteins, as well as the cytokines midkine and pleiotrophin, in human postheparin plasma. AB - The heparin-releasable proteins are a group of proteins that are targeted to the endothelial surface by attachment to glycosaminoglycans and may have functions specific to the endothelium-blood interface. In this study, heparin-affinity chromatography of human postheparin plasma was used as a method to identify and study novel heparin-releasable proteins. Six proteins seen on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels have increased levels in plasma after intravenous heparin. The six proteins are platelet factor 4, midkine, pleiotrophin, and several novel proteins. Midkine and pleiotrophin are related cytokines that are developmentally regulated, neurotrophic, and mitogenic. Additional studies show that levels of midkine and pleiotrophin peak at 10 to 30 minutes after injection of heparin. Heparin-releasable midkine and pleiotrophin do not originate from blood cells or the kidney. Heparin-releasable midkine may originate from endothelial cells. Soft agar culture of an adenocarcinoma cell line (SW-13) demonstrates growth-stimulating activity similar to that described for pleiotrophin in the heparin-agarose eluate of postheparin plasma but not in the heparin-agarose eluate of preheparin plasma. It is concluded there are more heparin-releasable proteins than previously identified, including midkine and pleiotrophin, and that heparin-affinity chromatography of postheparin plasma is a useful technique for identifying novel heparin-releasable proteins. PMID- 8241101 TI - Effects of local geometry and fluid dynamics on regional platelet deposition on artificial surfaces. AB - An important aspect of blood-material interactions is the activation, adhesion, and subsequent aggregation of blood platelets on the artificial surface, all of which are directly affected by local fluid dynamics. The objective of this work was to directly correlate changing local fluid dynamic conditions produced by various vessel geometries, including stenosis, aneurysm, and separate contraction and expansion geometries, with quantitative in vitro measurements of regional platelet deposition. We directly measured platelet deposition as a function of axial position along four Lexan flow chambers with axisymmetric models of these geometries using 111In-labeled platelets. Platelet deposition was maximum in observed areas of flow recirculation and reattachment and minimum in locations of high shear and separation. For the stenosis geometry, two distinct regions of increased platelet deposition were apparent, one proximal to and one distal to the stenosis throat. An approximately linear increase in platelet densities was produced in the aneurysm region, increasing in the direction of flow. Through a comparison of platelet deposition with local fluid streamline orientation, we have shown that platelet deposition is increased in certain areas due to the enhanced convective transport of platelets and blood cells to the vessel wall along locally curved streamlines with velocity components perpendicular to the vessel wall. PMID- 8241102 TI - Lack of apoA-I is not associated with increased susceptibility to atherosclerosis in mice. AB - The consequences of the lack of apolipoprotein A-I (apoA-I) were evaluated in mice made to lack apoA-I by gene targeting. Inbred strain 129 mice homozygous for the inactive Apoa1 gene and maintained on regular mouse chow had markedly reduced total cholesterol (26% normal) and high-density lipoprotein (HDL) cholesterol (25% normal) levels in their plasma. Their plasma lipoproteins lacked apoA-I and were reduced in all other apolipoproteins but apoE. ApoE comprises about one third of the protein of HDL particles in homozygotes, whereas it is present in only trace amounts in normal HDL. Despite the reduction of HDL cholesterol, no atherosclerotic lesions were observed in any of the homozygous mice evaluated (up to 15 months of age). After being maintained on an atherogenic diet for 4 weeks, total plasma cholesterol of the homozygous mutants increased by 20 mg/dL, while that of normals increased by 60 mg/dL. Mice with mixed 129 and C57BL/6J genetic backgrounds were fed the atherogenic diet for 20 weeks. A small number of foam cells were found attached to the aortic surface in some of the animals, but the extent and occurrence of these depositions were not related to the apoA-I genotype. Our results demonstrate that a lack of apoA-I does not by itself cause atherosclerosis in mice. PMID- 8241103 TI - Augmentation of synthesis of plasminogen activator inhibitor type-1 in arterial endothelial cells by glucose and its implications for local fibrinolysis. AB - Because of the frequent occurrence of premature cardiovascular disease in patients with non-insulin-dependent, type II diabetes mellitus (NIDDM), the attenuated fibrinolytic activity of plasma from type II diabetic patients with increased concentrations of plasminogen activator inhibitor type-1 (PAI-1), and the fact that insulin stimulates synthesis of PAI-1 by human hepatic cells in vitro, we and others have hypothesized that accelerated vascular disease in type II diabetes may result in part from impaired fibrinolysis secondary to excessive elaboration of PAI-1 stimulated by insulin. Alternatively, the hyperglycemia associated with type II diabetes could influence the synthesis and secretion of PAI-1 directly. The present study was performed to determine whether PAI-1 secretion is or is not sensitive to the prevailing concentration of glucose in the conditioned medium of endothelial and liver cells, which are thought to be the major sources of circulating PAI-1 in vivo. Confluent cells were exposed to 0, 2.8, 5.6, 11.1, or 22.2 mmol/L (0, 50, 100, 200, or 400 mg/dL) glucose in medium without serum and subsequently to media with or without insulin (7.3 nmol/L). Secretion of PAI-1 by highly differentiated human hepatoma (Hep G2) cells did not increase as a function of increasing concentrations of glucose, whether or not insulin was present. In contrast, with pig aortic endothelial cells, the secretion of PAI-1 increased significantly with extracellular glucose with or without insulin. The increases in PAI-1 were specific (as shown by metabolic labeling experiments) and not attributable to osmotic effects (as shown by replacement of glucose by sorbitol).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241104 TI - Association of hemostatic variables with prevalent cardiovascular disease and asymptomatic carotid artery atherosclerosis. The Atherosclerosis Risk in Communities (ARIC) Study Investigators. AB - The relation of hemostatic factor levels to the occurrence of cardiovascular disease is incompletely established. The Atherosclerosis Risk in Communities Study measured fibrinogen, factor VII, factor VIII, von Willebrand factor, antithrombin III, protein C, activated partial thromboplastin time, and other cardiovascular risk factors in nearly 15,000 men and women aged 45 to 64. This analysis assessed the relations of these hemostatic factors with prevalent cardiovascular disease and asymptomatic carotid artery intimal-medial thickness measured by B-mode ultrasound. Compared with participants without cardiovascular disease, those with cardiovascular disease had higher levels of fibrinogen, factor VIII, and von Willebrand factor in both sexes. The other hemostatic factors were less consistently associated with prevalent cardiovascular disease. Only fibrinogen was associated with carotid intimal-medial thickness. Adjusted for age, race, and field center, the odds ratio for carotid wall thickness in the 90th percentile or greater, compared with < 50th percentile, for each SD higher fibrinogen concentration (65 mg/dL) was 1.42 (95% confidence interval, 1.25, 1.62) in men and 1.43 (1.25, 1.64) in women. This population-based study provides further evidence that fibrinogen and possibly factor VIII and von Willebrand factor are risk factors for cardiovascular disease. PMID- 8241105 TI - A chimeric murine/human antibody Fab fragment directed against the platelet GPIIb/IIIa receptor enhances and sustains arterial thrombolysis with recombinant tissue-type plasminogen activator in baboons. AB - Inhibition of the platelet glycoprotein (GP) IIb/IIIa receptor with the murine monoclonal antibody 7E3 abolishes ex vivo platelet aggregation, reduces thrombogenicity, and sustains arterial recanalization with recombinant tissue type plasminogen activator (rt-PA). A chimeric murine/human Fab fragment of 7E3 (c7E3-Fab) has a markedly reduced immunogenicity, but its potency as an adjunct for thrombolysis with rt-PA has not been evaluated. The effects of a single intravenous bolus injection of aspirin (17 mg/kg) or c7E3-Fab (0.45 mg/kg) on thrombolysis and reocclusion induced with rt-PA were studied in groups of six baboons with femoral arterial thrombosis and superimposed high-grade stenosis. This dose of c7E3-Fab blocked 96 +/- 1% of the platelet GPIIb/IIIa receptors and abolished ADP-induced platelet aggregation. Bolus intravenous injections of rt-PA (0.25 mg/kg) were repeated at 15-minute intervals until reperfusion occurred (maximum of four injections). In the aspirin group, reperfusion was obtained within 51 +/- 16 minutes (mean +/- SD) but was rapidly followed by reocclusion within 6 +/- 9 minutes and by cyclic reflow and reocclusion. In the c7E3-Fab group, reperfusion was obtained within 25 +/- 8 minutes (P < .01 versus aspirin group) and was associated with a delayed reocclusion of 63 +/- 63 minutes (P < .05 versus aspirin group). Template bleeding times remained unchanged in the aspirin/rt-PA group but were markedly prolonged (to > 30 minutes) in the c7E3 Fab/rt-PA group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241106 TI - Determination of lipid transfer inhibitor protein activity in human lipoprotein deficient plasma. AB - Lipid transfer protein (LTP) activity is modulated by a distinct plasma protein termed lipid transfer inhibitor protein (LTIP). The objective of this study was to establish an assay for LTIP that could be used to quantify its activity in lipoprotein-deficient plasma. A straightforward heating protocol (56 degrees C for 60 minutes) was found to inactivate more than 90% of LTIP activity. The responses of individual lipoprotein-deficient plasma samples to this heating procedure were unique. Among normolipidemic donors, inactivation of LTIP caused a 230% to 600% increase in LTP activity. Essentially all measurable transfer activity in native and heated samples was inhibited by an antibody to LTP. Whole plasma samples from these donors were spiked with radiolabeled lipoproteins to measure the rates of lipid transfer among the major lipoprotein classes. In general, plasma lipid transfer rates were negatively correlated with LTIP activity in these samples. However, the decrease in lipid transfers from very-low density lipoprotein (VLDL) to low-density lipoprotein (LDL) and from LDL to VLDL was from 2.4- to 5.1-fold greater than in the transfers from VLDL to high-density lipoprotein (HDL) or from HDL to VLDL. In these samples, the molecular weight of HDL2 was negatively correlated with LTIP activity. Thus, LTIP activities among normolipidemic individuals were observed to vary severalfold; compared with other lipoprotein transfers, higher LTIP activities were associated with a relative reduction in LDL-VLDL lipid transfer events. PMID- 8241107 TI - Organization of von Willebrand factor on surface-activated platelets. AB - The distribution and organization of von Willebrand factor (vWF) multimers on platelets after surface activation have not been fully characterized. In the present study, washed human platelets were allowed to interact with Formvar coated, electron microscope grids for 20 minutes at 37 degrees C and then fixed. After fixation, cells were washed and then incubated with buffer alone, human plasma, human plasma preincubated with ristocetin (1.2 mg/mL), purified human vWF plus ristocetin, or bovine plasma. Macromolecular complexes were revealed by ultrastructural immunocytochemistry employing a polyclonal antibody against vWF and protein A-gold (PAG) as the electron-dense probe. vWF multimers were not present in discoid platelets but appeared on the central zone of dendritic cells and over larger central areas of fully spread platelets. Exposure to human plasma alone did not affect the distribution of electron-dense probes for vWF in central regions of surface-activated cells. Incubation of spread platelets with ristocetin-activated human plasma or bovine plasma resulted in the appearance of randomly dispersed, mottled areas of increased density covering the surface from edge to edge. Exposure to vWF antibody and PAG resulted in specific labeling of the dense areas in a serpentine, linear array. The gold-probe distribution suggested that the vWF multimers were not superimposed and were distributed in a random, irregular manner from edge to edge with label-free, clear areas between them. The results extend previous observations demonstrating that glycoprotein Ib IX receptors are not spontaneously cleared from the plasma membranes of surface activated platelets by showing that the receptor function of glycoprotein Ib-IX complex remains unchanged. PMID- 8241108 TI - Expression of follistatin, an activin-binding protein, in vascular smooth muscle cells and arteriosclerotic lesions. AB - Activin-A has a mitogenic effect on vascular smooth muscle cells (SMCs) and is produced by monocyte/macrophage lineage cells. Here, we studied the expression of follistatin, an activin-binding protein, in both A10 cells, a rat aortic SMC line, and vascular SMCs derived from adult rat aorta. Follistatin mRNA was detected in these cells by Northern blot analysis. Ligand blot and immunoblot analyses demonstrated that follistatin was produced in the conditioned medium at a higher level by A10 cells than by SMCs. Furthermore, immunostaining and in situ hybridization of the arteriosclerotic lesions showed that follistatin was highly expressed in the diseased artery, where abnormal proliferation of SMCs occurred. We suggest that follistatin is produced by vascular SMCs and is involved in the course of atherogenesis. PMID- 8241109 TI - Involvement of the hemostatic system in the insulin resistance syndrome. A study of 1500 patients with angina pectoris. The ECAT Angina Pectoris Study Group. AB - Hyperinsulinemia, a major indicator of insulin resistance, may exert its influence on the risk of coronary artery disease partially through disturbances of the hemostatic system. The relations of fasting insulin concentrations with the degree of coronary atherosclerosis, other coronary risk factors (including some markers of the insulin resistance syndrome such as body mass index and triglyceride), markers of inflammation, and hemostatic factors were investigated in 1484 patients with angina pectoris. Mean insulin levels were higher in patients with one or more coronary vessel stenoses than in those without (9.9 microU/mL compared with 9.0 microU/mL, P < .0001). However, the association the presence of vessel stenoses was stronger in patients with a previous myocardial infarction than in those without. Insulin increased markedly (P < .0001) and independently of other risk factors with age body mass index, triglyceride concentration, and markers of inflammation, such as white blood cell count and C reactive protein. The strongest relations between insulin and hemostatic factors were observed with fibrinolytic variables, particularly plasminogen activator inhibitor-1 (PAI-1) levels (r = .44, P < .0001). This relation decreased somewhat (r = .29) after simultaneous adjustment for markers of the insulin resistance syndrome, mainly body mass index and triglycerides, but not after adjustment for markers of inflammation. Therefore, we propose that increased PAI-1 levels, which are essentially related to the classic metabolic aspect of the insulin resistance syndrome, have to be included in this syndrome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241110 TI - The endothelium during cuff-induced neointima formation in the rabbit carotid artery. AB - Intimal thickening in human arteries is considered as a site of predilection for atherosclerosis. The placement of a flexible, physically nonconstrictive, silicone cuff around the rabbit carotid artery induced a neointima composed of smooth muscle cells (SMCs) within 14 days. To investigate possible alterations of the endothelial cells (ECs) during neointima formation, their morphology was examined with scanning electron microscopy (SEM), transmission electron microscopy (TEM), and confocal microscopy. In the early postoperative period (6 hours), both cuffed and sham-operated arteries demonstrated small foci (5 to 200 microns) of denudation, presumably as a consequence of the manipulation. Within 24 hours the luminal surface of the cuffed and sham-operated arteries was completely covered with endothelium, which remained continuous throughout the study. However, after 1 week the ECs of the cuffed arteries contained a pronounced rough endoplasmic reticulum. From 6 hours until 3 days, polymorphonuclear leukocytes infiltrated the cuffed but not the sham-operated arteries from the lumen. Subendothelial SMC accumulation in the cuffed arteries began after this time period. At day 14 a full-blown neointima composed of longitudinally oriented SMCs had formed in the cuffed arteries. The sham-operated arteries did not develop a neointima. During neointima formation immunoreactivity for von Willebrand factor (vWf) increased in the ECs, and vWf was deposited in the extracellular spaces of the neointima. At day 14 the area of vWf deposits correlated positively with the area of the neointima (r = .73, P < .001). In subsequent weeks, the intimal area did not increase, and vWf deposits vanished from the neointimal matrix. The endothelium of the sham-operated arteries showed no change in vWf immunoreactivity compared with untreated arteries throughout the study. The altered ultrastructural morphology of the ECs and the concurrent vWf deposition in cuffed but not in sham-operated arteries point to alterations in EC function during the development of the neointima. The vWf secretion could possibly lead to increased adhesiveness of the extracellular matrix for the ECs as well as modulate neointima formation. PMID- 8241111 TI - Effect of vitamin E on vascular integrity in cholesterol-fed guinea pigs. AB - This study was designed to clarify the effects of vitamin E on the alterations in proteoglycan distribution and vascular permeability, which were examined by immunohistochemical and ultrastructural techniques in the aortas of cholesterol fed guinea pigs. The animals were divided into three groups: a control group, a cholesterol group, and a vitamin E group. Serum levels of cholesterol, triglyceride, low-density lipoprotein, high-density lipoprotein, and thiobarbituric acid-reactive substances were measured. An increase in thiobarbituric acid-reactive substances was observed in the cholesterol group compared with control and vitamin E groups. Intimal atheromatous lesions of the aorta were significantly decreased in the vitamin E group compared with the cholesterol group. Histochemically, an increased distribution of proteoglycans such as chondroitin, dermatan, and heparan sulfates and ruthenium red reaction products in the intima; decreased glycocalyx on the endothelial surface; and increased permeability to horseradish peroxidase were revealed in the cholesterol group compared with the vitamin E group. Hypercholesterolemia, resulting in superoxide production, may have contributed to the endothelial damage and increased permeability to plasma proteins and lipids in the vascular wall of the cholesterol group. However, vitamin E administration inhibited lipid deposition and development of this abnormal permeability associated with an irregular distribution of proteoglycan. These results suggest that vitamin E preserves the morphological and functional integrity of the vascular wall and may contribute to the inhibition of atherogenesis in cholesterol-fed guinea pigs. PMID- 8241112 TI - Use of site-directed fluorescence labeling to study proximity relationships in the lactose permease of Escherichia coli. AB - The lactose permease of Escherichia coli is a paradigm for polytopic membrane transport proteins that transduce free energy stored in an electrochemical ion gradient into work in the form of a concentration gradient. Although the permease consists of 12 hydrophobic transmembrane domains in probable alpha-helical conformation that traverse the membrane in zigzag fashion connected by hydrophilic "loops", little information is available regarding the folded tertiary structure of the molecule. In this paper, we describe an approach to studying proximity relationships in lactose permease that is based upon site directed pyrene labeling of combinations of paired Cys replacements in a mutant devoid of Cys residues. Since pyrene exhibits excimer fluorescence if two molecules are within about 3.5 A, the proximity between paired labeled residues can be determined. The results demonstrate that putative helices VIII and IX are close to helix X. Taken together with other findings indicating that helix VII is close to helices X and XI, the data lead to a model that describes the packing of helices VII-XI. PMID- 8241113 TI - Specific ammonium ion requirement for functional ribosomal RNA tertiary structure. AB - In compactly folded RNAs, coordination or hydrogen bonding of cations in specific sites is a potentially important aspect of the tertiary structure. NH4+ specifically stabilizes the tertiary structure of a conserved, 58-nt fragment of the large subunit ribosomal RNA, as judged in two ways: a melting transition associated with tertiary interactions is sharpened and stabilized more effectively by NH4+ than by any alkali metal cation, and the affinity of the RNA fragment for ribosomal protein L11 or the antibiotic thiostrepton is approximately 10-fold stronger when measured in NH4+ than in Na+. The dependence of the melting temperature on NH4+ concentration shows that a single bound ion is responsible for these effects. The requirement of different ribosome functions for NH4+ suggests that other such sites exist in ribosomal RNAs. PMID- 8241114 TI - Pyrularia thionin binding to and the role of tryptophan-8 in the enhancement of phosphatidylserine domains in erythrocyte membranes. AB - Pyrularia thionin is a small, strongly basic peptide which interacts readily with cellular and synthetic membranes. With cells it induces hemolysis, depolarizes the cellular membrane with an accompanying influx of Ca2+, and activates an endogenous phospholipase A2. Evidence points toward a binding site involving phosphatidylserine (PS). This study shows that addition of the peptide to erythrocyte membranes as well as to vesicles formed from phospholipids isolated from erythrocyte membranes causes an enhancement of phospholipid domains which are made visible by the use of fluorescence digital imaging microscopy with fluorescent derivatives of PS (NBD-PS) and phosphatidylcholine (NBD-PC). Addition of thionin caused a large increase in NBD-PS domains, with an accompanying enrichment of NBD-PC in another separate domain. Double-labeling experiments performed with a Texas Red derivative of thionin show that the peptide binds to the domain enriched in NBD-PS. P thionin inactivated by modification of Trp-8 with N-bromosuccinimide lost the ability to enhance PS domains, although it bound to the membrane with the same affinity as native P thionin. This shows that binding to the membrane is not in itself sufficient to cause the NBD-PS and NBD PC redistribution into domains. PMID- 8241115 TI - NMR solution structure of an alpha-bungarotoxin/nicotinic receptor peptide complex. AB - We report the two-dimensional nuclear magnetic resonance (NMR) characterization of the stoichiometric complex formed between the snake venom-derived long alpha neurotoxin, alpha-bungarotoxin (BGTX), and a synthetic dodecapeptide (alpha 185 196) corresponding to a functionally important region on the alpha-subunit of the nicotinic acetylcholine receptor (nAChR) obtained from Torpedo californica electric organ tissue. BGTX has been widely used as the classic nicotinic competitive antagonist for the skeletal muscle type of nAChR which is found in the avian, amphibian, and mammalian neuromuscular junction. The receptor dodecapeptide (alpha 185-196) binds BGTX with micromolar affinity and has been shown to represent the major determinant of BGTX binding to the isolated alpha subunit. Previous studies involving covalent modification of the native nAChR from Torpedo membranes with a variety of affinity reagents indicate that several residues contained within the dodecapeptide sequence (namely, Tyr-190, Cys-192, and Cys-193) apparently contribute directly to the formation of the cholinergic ligand binding site. The NMR-derived solution structure of the BGTX/receptor peptide complex defines a relatively extended conformation for a major segment of the "bound" dodecapeptide. These structural studies also reveal a previously unpredicted receptor binding cleft within BGTX and suggest that BGTX undergoes a conformational change upon peptide binding. If, as we hypothesize, the identified intermolecular contacts in the BGTX/receptor peptide complex describe a portion of the contact zone between BGTX and native receptor, then the structural data would suggest that alpha-subunit residues 186-190 are on the extracellular surface of the receptor. PMID- 8241116 TI - Characterization of a folding intermediate of apoplastocyanin trapped by proline isomerization. AB - The unfolding and refolding transitions of French bean apoplastocyanin (apo-Pc), a beta-sandwich protein, have been characterized. The apoprotein is stabilized by sodium sulfate and can be reversibly unfolded by guanidine hydrochloride (GuHCl). However, in contrast to holo-Pc, apo-Pc is unstable at low ionic strength, suggesting that the copper ion stabilizes the holoprotein. The equilibrium unfolding transition monitored by peptide circular dichroism (CD) and tyrosine fluorescence is described by a two-state model. The kinetics of the unfolding transition were monitored using a manual mixing technique and are consistent with a single two-state transition. In contrast, the kinetics of the refolding reaction measured by fluorescence and CD show two transitions with different rates. The relaxation time of the slower phase (800-1000 s) is almost independent of GuHCl concentration. The faster phase was observed only under strongly native conditions, and its relaxation time is GuHCl-dependent. Double-jump experiments and acceleration by cyclophilin demonstrate that both phases involve cis-trans isomerization of proline residues. The changes in fluorescence associated with the two phases are more than 150% of the total change expected from equilibrium experiments, indicating the presence of intermediate(s) with fluorescence greater than the unfolded state. Amide hydrogen-exchange experiments coupled with two dimensional NMR spectroscopy demonstrate the formation of an intermediate in the very low refolding reaction in which amide protons in the beta-sheets are weakly protected from exchange. No CD evidence for nativelike beta-sheet formation was found for this intermediate. The NMR experiments suggest that the intermediate is compact with flexible beta-sheets and altered packing of the hydrophobic core. It has many of the characteristics of a molten globule. However, the 1H NMR spectrum of the intermediate exhibits a small number of shifted resonances that indicate the presence of specific tertiary interactions in a localized region. A mechanism for refolding of apoplastocyanin is proposed that includes two slow steps corresponding to trans-->cis isomerization of two prolines. PMID- 8241117 TI - Circular permutation of T4 lysozyme. AB - To examine the relationship between polypeptide chain synthesis and protein folding, we have constructed a circularly permuted variant of phage T4 lysozyme. The permuted protein begins at residue 37 of the wild-type sequence and ends at residue 36. The normal chain termini are joined by a six-residue linker, Ser-Gly4 Ala. The permuted lysozyme folds efficiently and cleaves bacterial cell walls with normal specific activity. As judged by circular dichroism, UV absorbance, fluorescence, and nuclear magnetic resonance spectroscopy, the permutation causes little change in the structure of the protein. Reversible denaturation experiments show that the permutation reduces the stability of T4 lysozyme only 0.8-1.1 kcal/mol. These results demonstrate that a protein with two domains can be permuted with little change in activity, structure, and stability. The order of chain synthesis, the sequential arrangement of secondary structures, and the position of chain termini with respect to domain boundaries do not determine the protein fold. PMID- 8241118 TI - Adiabatic compressibility of molten globules. AB - We report the measurement of the adiabatic compressibility beta for "molten globule" states of cytochrome b562 and cytochrome c. Precise density and sound velocity measurements allow determination of beta in aqueous solution. Surprisingly, the molten globule apocytochrome b562 shows a beta comparable to the native holocytochrome b562. We estimated the maximum difference in the hydration contribution to the compressibility of apocytochrome b562 relative to holocytochrome b562 to be only -2.6 x 10(-11) Pa-1. Our results suggest that the intrinsic compressibility and the overall volume fluctuations in the molten globule state differ only slightly from those in the native protein. With cytochrome c we investigated the changes in beta for the transitions from the native to the acid molten globule and expanded premolten globule states. The molten globule at pH 2.2 was found to be somewhat "softer" than the native protein at neutral pH. In contrast, a decrease in beta of the premolten globule indicates an increased penetration of water into the macromolecule. In both of these intermediates the differences in adiabatic compressibility relative to the native state are small in comparison to the range of adiabatic compressibility observed for different native globular proteins. PMID- 8241119 TI - Kinetics and free energy gaps of electron-transfer reactions in Rhodobacter sphaeroides reaction centers. AB - The rates of the light-driven, electron-transfer reactions in the photosynthetic reaction center (RC) of Rhodobacter sphaeroides are examined in mutant strains in which tyrosine (M)210 is replaced by phenylalanine, isoleucine, or tryptophan. The spectra of the absorbance changes between 700 and 975 nm, following excitation by 0.6-ps pulses at 605 nm, are analyzed globally by singular value decomposition. The spectra measured at room temperature are interpreted in terms of a model in which the excited bacteriochlorophyll dimer (P*) transfers an electron to a bacteriopheophytin (HL) with time constants of 3.5 +/- 0.3, 10.5 +/ 1.0, 16 +/- 2, and 41 +/- 4 ps in wild-type RCs and the Phe, Ile, and Trp mutants, respectively, and an electron then moves from HL- to a quinone (QA) with a time constant of 0.16 ns in wild-type RCs, 0.24 ns in the Phe mutant, and 0.20 ns in the Ile and Trp mutants. The first step speeds up with decreasing temperature in wild-type RCs, remains virtually unchanged in the Phe mutant, and slows down in the Ile and Trp mutants. At 80 K, the signals in the 850-975-nm region include an apparent shift of the stimulated emission or absorption spectrum of P*, with a time constant of 5 ps in the Ile mutant and 13 pcs in the Trp mutant. Most of the electron transfer to HL occurs with time constants of 55 and 155 ps in the Ile and Trp mutants, respectively, and probably occurs from the relaxed form of P*. Electron transfer from the initial state cannot be ruled out, however. Relaxations of P* are not resolved in wild-type RCs or the Phe mutant. The midpoint potential (Em) of the P/P+ redox couple is measured by an electrochemical technique; the Em values are 500 +/- 5, 530 +/- 6, 533 +/- 3, and 552 +/- 10 mV for the wild-type and the Phe, Ile, and Trp mutant RCs, respectively. These values are corroborated by chemical titrations. The free energy change (delta G degrees) associated with formation of the P+HL-radical pair from P* also is determined by measuring the amplitude of fluorescence on the nanosecond time scale after blocking electron transfer from HL- to QA. The free energy of P+HL- is elevated by an amount comparable to that calculated from the increase in the Em of P in the Ile mutant and by about 16 meV more than this in the Phe and Trp mutants.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8241120 TI - Fluorescence study of conformational flexibility of RNase S-peptide: distance distribution, end-to-end diffusion, and anisotropy decays. AB - Frequency-domain fluorescence resonance energy transfer and anisotropy measurements were performed to characterize conformational dynamics of an analog of the RNase S-peptide (residues 1-20). Trp was used as a donor by replacing Phe 8, and a dansyl acceptor group was introduced at position 1 or 18. The distance distribution parameters, half width of the distribution, end-to-end diffusion coefficient, and to some extent anisotropy decays were sensitive to changes in the S-peptide conformation. The observed mean distance of about 13-14 A between residues 1 and 8 in the presence of 50% TFE and when bound to RNase S-protein is in reasonable accord with the X-ray structure of RNase. The mean distance of 9.3 A between residues 8 and 18 in the presence of 50% TFE is, however, significantly smaller than 15.3 A found for the S-protein complex. The half-width of the distance distribution increased from about 9 to 18 A for residues 1-8 and from about 6 to 14 A for segment 8-18 with the loss of helical structure. The half widths of 9 A in the case of 1-8 segment when peptide is helical suggests the presence of considerable conformational heterogeneity. Also, the 14 A half-width for segment 8-18 when it is random-coil is smaller than that expected for a random coil 11-residue segment. The donor-to-acceptor diffusion coefficients were less than 1 x 10(-7) cm2/s at 2 degrees C for both segments and increased to 1-2 x 10(-6) cm2/s at 35 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241121 TI - Formation of liquid crystals from actin filaments. AB - Actin is cross-linked by actin-binding proteins in the cytoplasm to form either isotropic or highly oriented anisotropic structures. The inherent orientation among actin filaments could influence whether an isotropic or highly oriented anisotropic structure is formed. A highly oriented state can arise spontaneously through the formation of liquid crystals as predicted by polymer theory. In this study, the ability of filamentous actin to form liquid crystalline domains was detected using the anisotropic component of scattered light and by observation of birefringence. As liquid crystalline domains formed, the intensity of the anisotropic component of scattered light increased, and birefringent macroscopic oriented domains were directly observed. The formation of liquid crystalline domains was dependent on the concentration of actin filaments and on the average filament length controlled by varying the ratio of gelsolin to actin monomers. The concentration of actin filaments required to form liquid crystalline domains increased moderately as the average length was decreased. At a fixed actin concentration, orientation among the filaments attained a maximum value at a ratio of actin to gelsolin in the range from 1500 to 2000 and decreased as the ratio was increased or decreased from this range. The results are not well explained by theoretical treatments for liquid crystal formation by monodisperse, charged worm-like chains. Differences from the theoretical predictions for formation of liquid crystals are most likely due to the polydisperse filament length of actin. This phenomenon may have important effects on the structural and rheological properties of the cytoplasm in living cells. PMID- 8241122 TI - Actin filament annealing in the presence of ATP and phalloidin. AB - The re-formation of actin filaments after fragmentation by sonication in the presence of phalloidin and ATP has been found to follow second-order kinetics. The data are described by a model in which the rate of actin filament annealing is proportional to the square of the number concentration of actin filaments and the rate of fragmentation is proportional to the actin polymer concentration. In the presence of 100 mM KCl, 1 mM MgCl2, and equimolar phalloidin, the second order rate constant for annealing of actin filaments is 2.2 x 10(6) M-1 s-1 and the first-order rate constant for fragmentation is 7 x 10(-7) s-1. In addition, the observed pseudo-first-order rate constant for annealing was found to increase with increasing ionic strength. Thus, annealing may play a major part in the length redistribution phase of actin polymerization and may be important for actin filament rearrangement in the cell. PMID- 8241123 TI - Stimulation of Ca(2+)-regulated olfactory phospholipase C by amino acids. AB - L-Amino acids are potent olfactory stimuli for Atlantic salmon. A plasma membrane fraction, previously shown to be rich in amino acid binding sites, was prepared from olfactory rosettes of Atlantic salmon (Salmo salar) and utilized to investigate the role of phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis in olfactory signal transduction. A cocktail of L-amino acids (Ser, Glu, Lys, and Gly) stimulated PIP2 hydrolysis by phospholipase C (PLC) in a dose-dependent manner with half-maximal stimulation when all amino acids were present at approximately 1 microM. Stimulation of PIP2 hydrolysis by amino acids required GTP gamma S, which alone had no effect on PLC activity. Unlike GTP gamma S, AlF4- and Ca2+ stimulated PIP2 breakdown. Preincubation with 1 mM GDP beta S eliminated the effect of amino acids and AlF4- on PIP2 hydrolysis, suggesting the involvement of G protein regulation. The lack of stimulation by GTP gamma S alone suggested that there was negligible exchange of GTP gamma S for GDP in the absence of odorant. There were no significant effects of amino acids on either adenylate cyclase or guanylate cyclase activities in the membrane preparation under these conditions. The effect of the amino acid cocktail was maximal at 1-10 nM free Ca2+. At or above 100 nM free Ca2+, no effect of amino acids on PIP2 hydrolysis was found. However, between 100 nM and 100 microM, Ca2+ directly stimulated PLC activity in a dose-dependent manner. This stimulation by Ca2+ appeared to be G protein independent because it did not require GTP gamma S and was not inhibited by GDP beta S.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241124 TI - Structural characterization, membrane interaction, and specific assembly within phospholipid membranes of hydrophobic segments from Bacillus thuringiensis var. israelensis cytolytic toxin. AB - The Bacillus thuringiensis var. israelensis (Bti) cytolytic toxin is hypothesized to exert its toxic activity via pore formation in the cell membrane as a result of the aggregation of several monomers. To gain insight into the toxin's mode of action, 2 putative hydrophobic 22 amino acid peptides were synthesized and characterized spectroscopically and functionally. One peptide corresponded to the putative amphiphilic alpha-helical region (amino acids 110-131, termed helix-2), and the other to amino acids 50-71 (termed helix-1) [Ward, E. S., Ellar, D. J., & Chilcott, C. N. (1988) J. Mol. Biol. 202, 527-535] of the toxin. Circular dichroism spectroscopy revealed that both segments adopt high alpha-helical content in a hydrophobic environment, in agreement with previous models. To monitor peptide-lipid and peptide-peptide interactions, the peptides were labeled selectively with either 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD) (to serve as donor) or tetramethylrhodamine (to serve as an acceptor), at their N-terminal amino acids. Both segments bind strongly to small unilamellar vesicles, composed of zwitterionic phospholipids, with surface partition coefficients on the order of 10(4) M-1. The shape of the binding isotherms indicates that helix-2 forms large aggregates within phospholipid membranes. Resonance energy transfer experiments demonstrated that the segments self-associate and interact with each other, but do not associate with unrelated membrane-bound peptides. Functional characterization demonstrated that helix-2 permeates phospholipid SUV with a potency similar to that of naturally occurring pore-forming peptides. Thus, the results support a role for helices-1 and -2 in the assembly and in the pore formation by Bti toxin. PMID- 8241125 TI - Biochemical characterization of the three major subclasses of lipoprotein A-I preparatively isolated from human plasma. AB - Apolipoprotein (apo) A-I is the major protein constituent of plasma high-density lipoproteins (HDL). HDL consist of two major classes of apoA-I-containing lipoproteins: LpA-I and LpA-I:A-II. LpA-I includes heterogeneous lipoprotein particles that differ in size and hydrated density. LpA-I was isolated by immunoaffinity chromatography from the fasting plasma of 24 normal human subjects and separated by gel filtration chromatography. Three major subclasses of LpA-I were eluted: large (Lg-LpA-I), medium (Md-LpA-I), and small LpA-I (Sm-LpA-I). By nondenaturing gradient PAGE, Lg-LpA-I, Md-LpA-I, and Sm-LpA-I had mean Strokes diameters of 10.8 +/- 0.5, 8.9 +/- 0.5, and 7.5 +/- 0.3 nm, respectively. The lipid/protein ratios were 1.25 +/- 0.12 for Lg-LpA-I, 0.75 +/- 0.10 for Md-LpA-I, and 0.38 +/- 0.08 for Sm-LpA-I. Lg-LpA-I was relatively lipid and cholesteryl ester rich compared with Md-LpA-I and Sm-LpA-I. Sm-LpA-I contained phospholipids as the major lipid component. ApoA-I was the major apolipoprotein in all LpA-I subfractions, whereas apoE was present only in Lg-LpA-I and apoA-IV was associated with both Md-LpA-I and Sm-LpA-I. All three LpA-I subclasses exhibited predominantly alpha mobility on agarose electrophoresis. Lg-LpA-I migrated as a diffuse band in the fast alpha position, whereas Md-LpA-I and Sm-LpA-I migrated to the slow alpha position.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241126 TI - Use of nitrogen-15 kinetic isotope effects to elucidate details of the chemical mechanism of human immunodeficiency virus 1 protease. AB - We have used 15N kinetic isotope effects of the HIV-1 protease-catalyzed peptidolysis of Ac-Ser-Gln-Asn-Tyr-Pro-Val-Val-NH2 to characterize the chemical mechanism of this enzyme. In addition, the multiple isotope effects have been determined by measuring the 15N kinetic isotope effects in both H2O and D2O. The isotope effects, measured on values of V/K, were determined by the incorporation of a radiolabel (tritium and 14C in peptides bearing the heavy and light isotopes, respectively) at a position remote from the isotopically labeled scissile peptide bond, such that the isotope effect was determined by measurement of the change in the 14C/3H ratio in recovered substrates at various fractions of reaction. At pH = 6.0 (37 degrees C), the nitrogen isotope effects were slightly, but significantly, inverse in both solvents: 15(V/K)H2O = 0.995 +/- 0.002, and 15(V/K)D2O = 0.992 +/- 0.003. The observation of an inverse nitrogen kinetic isotope effect implies that bonding to the nitrogen atom is becoming stiffened in a reaction transition state, and since this inverse isotope effect is enhanced in D2O, this isotope effect likely arises from protonation of the proline nitrogen atom.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241127 TI - Photoaffinity labeling of acyl-CoA oxidase with 12-azidooleoyl-CoA and 12-[(4 azidosalicyl)amino]dodecanoyl-CoA. AB - Synthesis of 32P-labeled CoA of high specific activity was achieved using partially purified dephospho-CoA kinase (EC 2.7.1.24) from pig liver with [gamma 32P]ATP as donor and dephospho-CoA as acceptor. A photoaffinity dodecanoic acid analog, 12-[(4-azidosalicyl)amino]dodecanoic acid was synthesized, as were its CoA derivative (ASD-CoA) and the CoA derivative of 12-azidooleic acid. The CoA derivatives were synthesized from azido fatty acid analogs by acyl-CoA synthetase. The synthesized photolabile reagents were tested as photoaffinity labels for acyl-CoA oxidase (EC 1.3.99.3) from Arthrobacter species. When a mixture of oxidase and the acyl-CoA analogs were incubated in the absence of ultraviolet light, the analogs were recognized as substrate. Acyl-CoA oxidase was incubated in the presence of acyl-CoA analogs and immediately photolyzed, which resulted in irreversible inhibition. Oleoyl-CoA and dodecanoyl-CoA protect the enzyme from photoactivated inhibition by 12-azidooleoyl-CoA and ASD-CoA, respectively. Analysis of photolyzed enzyme preparations by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed that both analogs preferentially labeled a 54,000 molecular weight protein. These results demonstrate that the photoaffinity acyl-CoA analogs have potential application as probes to identify and characterize lipid biosynthetic enzymes and to identify the active site of these proteins. PMID- 8241128 TI - Stereochemistry of 3-deoxyoctulosonate 8-phosphate synthase. AB - (Z)- and (E)-[3-2H]phosphoenolpyruvate were prepared chemically by the reductive deuteration of (Z)- and (E)-3-bromophosphoenolpyruvate, respectively, and were converted into 3-deoxyoctulosonate 8-phosphates deuterated at the C-3 position by incubation with unlabeled D-arabinose 5-phosphate in the presence of the enzyme, 3-deoxyoctulosonate 8-phosphate synthase (EC4.1.2.16) purified from Escherichia coli K-12 containing the plasmid pMW101. Analysis of the stereochemistry of the two 3-deoxyoctulosonate 8-phosphates deuterated at the C-3 position by 1H NMR showed that the (Z)-[3-2H]phosphoenolpyruvate had produced [3-2H]-3 deoxyoctulosonate 8-phosphate of predominantly the 3S configuration and that the E isomer had given predominantly (3R)-[3-2H]-3-deoxyoctulosonate 8-phosphate. The 3-deoxyoctulosonate 8-phosphate synthase reaction is therefore stereospecific with respect to the C-3 of phosphoenolpyruvate. The results indicate a si face attack from the C-3 of phosphoenolpyruvate, a result identical to that reported for 3-deoxyheptulosonate 7-phosphate synthase (EC 4.1.2.15), an enzyme catalyzing an identical aldol-type condensation, except that it takes place between phosphoenolpyruvate and D-erythrose 4-phosphate. The stereochemistry with respect to the face of the carbonyl of the attacked aldehyde, in both 3-deoxyoctulosonate 8-phosphate synthase and 3-deoxyheptulosonate 7-phosphate synthase, is re. On the basis of the results of the studies reported herein, the presence of a transient methyl group at the C-3 of phosphoenolpyruvate as part of the reaction mechanism seems unlikely. PMID- 8241129 TI - Flexibility impaired by mutations revealed the multifunctional roles of the loop in glutathione synthetase. AB - The loop from Ile-226 to Arg-241 in the glutathione synthetase (GSHase) from Escherichia coli B is rich in glycine and alanine and too flexible to take a fixed conformation [Yamaguchi, H., Kato, H., Hata, Y., Nishioka, T., Kimura, A., Oda, J., & Katsube, Y. (1993) J. Mol. Biol. 229, 1083-1100; Tanaka, T., Kato, H., Nishioka, T., & Oda, J. (1992) Biochemistry 31, 2259-2265]. To restrict the flexibility, three residues in the loop, Pro-227, Gly-229, and Gly-240, were replaced with alanine and valine residues. Variability in conformations of the mutant loops and shifts in the distribution of conformers between the open and closed states were assessed by steady-state kinetics, X-ray crystallographic structure analysis, and proteolysis with arginyl endopeptidase. Mutant enzymes replaced with a valine residue at the basal positions of the loop (P227V, G240V, and P227V/G240V) were identical with the wild-type enzyme in their crystal structures, except the loop region. The mutant loops retained apparent conformational variability, so as to take the open and closed states and to protect the acyl phosphate intermediate from the decomposition uncoupled from glutathione synthesis, but lost the catalytic activity; Kmapp values for glycine and gamma-Glu-Cys were sensitive to the mutations and drastically increased, and the k0app value was fatally reduced in the P227V/G240V mutant enzyme. The present results suggest that adjustability of the loop to the closed state is required for the recognition of the substrates, gamma-Glu-Cys and glycine, and for the chemical interactions with the bound substrates. PMID- 8241130 TI - Acidic residues are involved in substrate recognition by two soluble protein tyrosine phosphatases, PTP-5 and rrbPTP-1. AB - The mechanisms for substrate recognition by two cytoplasmic protein tyrosine phosphatases, PTP-5 and rrbPTP-1, were investigated. Phosphorylation sites on tyrosine-phosphorylated casein, a model PTP substrate, were characterized. Two peptides based on casein phosphorylation sites and one peptide based on the tyrosine phosphorylation site of reduced, carboxamidomethylated and maleylated (RCM) lysozyme were tested as PTP substrates. The three peptides were dephosphorylated by PTP-5 and rrbPTP-1 at rates comparable to those of the corresponding sites on the intact proteins. This indicates that peptides based on the two model PTP substrates, casein and RCM-lysozyme, contained all or most of the structural information necessary for PTP-5 and rrbPTP-1 substrate recognition. Structural elements required for substrate recognition by PTP-5 and rrbPTP-1 were also investigated. Km values for dephosphorylation of three simple aromatic phosphate esters (phosphotyrosine, p-nitrophenyl phosphate, and phenyl phosphate) by rrbPTP-1 were about 5000-fold higher than those obtained for the peptide and protein substrates. This indicates that recognition of protein and peptide substrates involves structural elements in addition to the phosphate group and the aromatic tyrosine ring of phosphotyrosine. Analysis of the effects of truncations and Ala for polar substitutions on the reactivity with PTP-5 and rrbPTP-1 of peptides based on casein, RCM-lysozyme, and angiotensin II indicated that Asp or Glu within the first five residues on the N-terminal side of phosphotyrosine increased peptide reactivity with both PTP's. Asn residues were unable or only weakly able to substitute for Asp residues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241131 TI - Protein-lipid interactions and Torpedo californica nicotinic acetylcholine receptor function. 1. Spatial disposition of cysteine residues in the gamma subunit analyzed by fluorescence-quenching and energy-transfer measurements. AB - The nicotinic acetylcholine receptor from Torpedo californica was labeled with a fluorescent, lipophilic probe, N-(1-pyrenyl)maleimide, specific for sulfhydryls in a hydrophobic environment, and was found to alkylate Cys 416, Cys 420 and Cys 451 in the gamma subunit [Li, L., Schuchard, M., Palma, A., Pradier, L., & McNamee, M.G. (1990) Biochemistry 29, 5428-5436]. The spatial disposition of the acetylcholine receptor-bound pyrene with respect to the membrane bilayer was assessed by a combination of fluorescence-quenching and resonance energy transfer measurements, under conditions of selective labeling of the gamma subunit. Quenching of pyrene fluorescence by spin-labeled fatty acids with the doxyl group at positions C-5 and C-12 revealed that the former was more effective, with a Stern-Volmer quenching constant of 0.187 compared to 0.072 for the latter, suggesting that the fluorophore(s) are located closer to the membrane-water interface rather than the hydrophobic interior. Energy transfer was found to occur from tryptophan in the acetylcholine receptor to cysteine-bound pyrene with a distance of separation of approximately 18 A. However, there was no energy transfer when pyrene-labeled AChR was reconstituted into membranes containing brominated phospholipids and cholesterol, suggesting that the fluorophore(s) responsible for energy transfer are located in the membrane domain. Thus, the N (1-pyrenyl)maleimide can be used to monitor lipid-protein interactions of the AChR. PMID- 8241132 TI - Protein-lipid interactions and Torpedo californica nicotinic acetylcholine receptor function. 2. Membrane fluidity and ligand-mediated alteration in the accessibility of gamma subunit cysteine residues to cholesterol. AB - Fluorescence-quenching and energy-transfer measurements were carried out to further characterize lipid-protein interactions involving the nicotinic acetylcholine receptor (AChR) from Torpedo californica in reconstituted membranes. To assess the fluidity of the receptor microenvironment, cis- and trans-parinaric acids were used to take advantage of the preferential partitioning behavior of the trans isomer for the gel phase. A relatively higher extent of energy transfer from the intrinsic tryptophan fluorescence of AChR in dielaidoylphosphatidylcholine bilayers to cis-parinaric acid in both the gel and the fluid phase suggests that the AChR is surrounded by a relatively fluid annulus of lipids. The ability of AChR to accommodate and interact with specific lipids such as cholesterol and fatty acids in the vicinity of pyrene-labeled cysteine residues in the membranous domain and/or the membrane-water interface region of the gamma subunit was assessed. Pyrene-labeled AChR prepared in (6,7 dibromostearoyl)phosphatidylcholine showed a 25% decrease in fluorescence as sites accessible to phospholipids were occupied; subsequent addition of dibromocholesterol hemisuccinate (DiBrCHS) caused further quenching by about 25%. This result is consistent with the presence of sites accessible to cholesterol, but not accessible to phospholipids, in the vicinity of the cysteine-bound pyrene in the membranous domain of the AChR. Quenching by DiBrCHS was sensitive to the presence of an AChR activator (carbamylcholine) but not a competitive antagonist (alpha-bungarotoxin). The Stern-Volmer quenching constant was 0.123 in the absence of added ligands and 0.167 and 0.134 in the presence of carbamylcholine and alpha-bungarotoxin, respectively, corresponding to accessibilities of 65%, 90%, and 70%. PMID- 8241133 TI - Purification and functional characterization of the ligand-binding domain from the retinoic acid receptor alpha: evidence that sulfhydryl groups are involved in ligand-receptor interactions. AB - The pGEX-2T expression vector was used to produce the ligand-binding domain from the human retinoic acid receptor alpha (hRAR alpha LBD) in Escherichia coli. The resulting fusion protein, containing the glutathione S-transferase separated from the truncated receptor (hRAR alpha 186-462) by a thrombin cleavage site, was purified with use of affinity chromatography on immobilized glutathione. A 90% homogeneity was obtained, with a specific activity of 100 pmol/mg and an overall 10% yield. Following purification and thrombin cleavage, a predominant monomeric (stokes radius = 2.3 nm, molecular mass of 32 kDa) [3H]retinoic acid hRAR alpha LBD complex was characterized by high-performance size-exclusion chromatography. The purified hRAR alpha LBD bound retinoic acid with an apparent Kd of 9 nM, a value close to the Kd of the full-length hRAR alpha expressed in COS cells. Kinetic studies at 0 degrees C demonstrate that the association of [3H]retinoic acid and [3H]CD367, a synthetic retinoid, to the overexpressed receptor was extremely rapid (complete in less than 3 min), whereas their dissociation from the receptor was slower, with half-lives of about 40 min at 0 degrees C. Experiments performed at various subzero temperatures allowed a more accurate assay of the association rate constant and indicate that the entropy of activation (delta Sa) is positive, which is characteristic of hydrophobic interactions. The ligand-binding activity was markedly decreased by pretreatment with various sulfhydryl modifying agents. 5,5'-Dithiobis(2-nitrobenzoic acid) (DTNB) appeared to be the most potent, whereas iodoacetamide was the least active. Furthermore, a series of N-alkylmaleimides was shown to inactivate the recombinant receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241134 TI - Structure and phase behavior of a charged glycolipid (1,2-O-dialkyl-3-O-beta-D glucuronosyl-sn-glycerol). AB - In order to investigate the effects of a net surface charge on the properties of glycolipid membranes, we have synthesized a glyceroglycolipid, 1,2-O-dialkyl-3-O beta-D-glucuronosyl- sn-glycerol (GlcUA lipid), with saturated alkyl chains of varying length (14, 16, and 18 carbon atoms; 14-, 16-, and 18-GlcUA, respectively) and glucuronic acid with an ionizable 6-carboxyl group as polar residue. Aqueous dispersions of GlcUA lipids have been characterized by differential scanning calorimetry, densitometry, and X-ray diffraction methods as a function of pH. The carboxyl group deprotonation of apparent pK about 5.5 leads to a decrease of the melting temperatures by about 7 degrees C for all three compounds and to a chain-length-dependent reduction of the transition enthalpies by 0, 7, and 14% for 14-, 16-, and 18-GlcUA, respectively. The decrease of the transition temperature is consistent with current electrostatic concepts and models of charged membrane interfaces, but the chain-length-specific dependence of the enthalpy decrease with an increase of pH shows that the pH effects in GlcUA lipids are not of purely electrostatic origin. However, these effects appear to be simpler in some instances than corresponding effects in phospholipids with multiply ionizable head groups. For this reason, the lipids with the glucuronic acid head group appear to represent an appropriate model system for studies of net electric charge effects on the membrane properties.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241135 TI - Secondary structure of M13 coat protein in phospholipids studied by circular dichroism, Raman, and Fourier transform infrared spectroscopy. AB - There is considerable uncertainty about the precise secondary structure adopted by the M13 coat protein when embedded in a phospholipid bilayer. Circular dichroism (CD) spectroscopy suggests that a major change in the structure of the coat protein occurs upon membrane insertion. It is reported that the structure of the protein in the membrane has only about 50% alpha-helix, the rest being mainly in a beta-sheet conformation, whereas the protein is almost completely alpha helical when intact in the phage. In this study we have undertaken a spectroscopic analysis using Fourier transform infrared, Raman, and CD spectroscopy to characterize the secondary structure of M13 coat protein when present in membranes consisting of dioleoylphosphatidylglycerol and dimyristoylphosphatidylglycerol. In sharp contrast to earlier CD studies, our results indicate that the coat protein in its membrane-embedded state has a very high alpha-helical content with virtually no beta-sheet structures present. This result indicates that the structures of the coat protein when intact in the phage or when embedded in the membrane are similar. Although our results differ from earlier CD studies, they are consistent with a recent NMR study, which showed that the M13 coat protein in sodium dodecyl sulfate micelles is primarily alpha helical with no evidence for beta-sheet structure [Henry, G. D., & Sykes, B.D. (1992) Biochemistry 31, 5284-5297]. These results lead to the conclusion that the M13 coat protein can insert from the membrane-bound state into a virus particle with a similar secondary structure, without large energy implications.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241136 TI - Resonance Raman spectroscopy of the azurin His117Gly mutant. Interconversion of type 1 and type 2 copper sites through exogenous ligands. AB - The copper center of the Pseudomonas aeruginosa His117Gly azurin mutant is accessible to exogenous ligands through an aperture in its surface created by the removal of the endogenous imidazole ligand. Depending on the exogenous ligand, a surprising variety of type 1 and type 2 copper sites can be obtained that are readily distinguished by electronic, EPR, and resonance Raman (RR) spectroscopy. The RR spectrum of type 1 H117G with exogenous imidazole is nearly identical to that of wild-type azurin, indicating that the trigonal geometry and short Cu S(Cys) bond of approximately 2.15 A have been maintained. With anionic ligands (e.g., Cl-, Br-, N3-), the RR spectra show increased intensity at 370 and 400 cm 1 and a corresponding decrease in intensity at 410 cm-1, suggesting a lengthening of the Cu-S(Cys) bond as the site achieves a more tetrahedral character. An extreme example is the hydroxide adduct of H117G which is green in color and has optical and RR spectra reminiscent of the tetrahedral type 1 site in Achromobacter cycloclastes nitrite reductase. The fact that the basic RR pattern is little changed in most of the type 1 adducts indicates that the RR spectrum is due primarily to vibrations of the Cu-cysteinate moiety and that its coplanar conformation is conserved. Type 2 H117G proteins are formed by the addition of bidentate exogenous ligands such as histidine and histamine. They have their absorption maxima blue-shifted to 400 nm and their EPR A parallel values increased to approximately 160 x 10(-4) cm-1, both of which are characteristic of tetragonal Cu sites with Cu-S(thiolate) bonds of > 2.25 A. The RR spectra of the type 2 H117G proteins are still dominated by multiple cysteinate-related vibrational modes. However, the vibrational modes with the greatest intensity and Cu-S(Cys) stretching character have shifted approximately 100 cm-1 to lower energy compared to the type 1 sites, consistent with a longer (Cys)S-Cu bond. It is proposed that the tetragonal type 2 character of the bidentate ligand complexes is due to the addition of a fourth strong ligand in the equatorial ligand plane. PMID- 8241137 TI - Electron transfer in sulfite oxidase: effects of pH and anions on transient kinetics. AB - Intramolecular electron transfer (ET) rates in sulfite oxidase (SO) were measured using flavin semiquinone reductants [5-deazariboflavin (dRFH.) and lumiflavin (LFH.)] generated by laser flash photolysis. Rapid bimolecular reduction of the heme by the dRF semiquinone occurred (k = 4 x 10(8) M-1 s-1 at pH 6; 1 x 10(8) M 1 s-1 at pH 9), followed by heme Fe(II) reoxidation due to intramolecular electron transfer to Mo(VI). Flash-induced difference spectra indicated that only spectral processes due to reduction and oxidation of the b-type heme prosthetic group were observed, with no detectable spectral contribution from the Mo cofactor. The extent of reoxidation decreased greatly from pH 6 to 9 (50% to 3%), as expected from the shifts in the redox potentials of the heme and Mo cofactor with pH, consistent with an electron transfer equilibrium between the two redox centers. The observed rate constant for the Fe(II) to Mo(VI) electron transfer decreased from 1650 s-1 at pH 6 to 60 s-1 at pH 9 and showed a maximum of 2400 s 1 at pH 7. Increases in salt concentration greatly decreased intramolecular ET rate constants (direct reduction by flavin semiquinone was unchanged), due to the binding of anions. Titration with the sodium salts of Cl-, SO4(2-), and H2PO4 /HPO4(2-) resulted in decreases in rate constants of intramolecular ET from 1500 s-1 to < 100 s-1 at pH 6 and 7. Similar dissociation constants were measured for the binding of these anions by flash photolysis and by steady-state enzyme kinetics using the inhibition of the sulfite/cytochrome c assay reaction for sulfite oxidase. A mechanism is proposed in which anion binding to the enzyme inhibits the rate of intramolecular electron transfer. PMID- 8241138 TI - Effect of alkylating agents on initiation and elongation of the lac UV5 promoter. AB - DNA containing the lac UV5 promoter was alkylated using bifunctional sulfur and nitrogen mustards and a monofunctional sulfur mustard. The alkylation sites were mapped using Taq polymerase, and the effect of alkylation on the formation of the DNA-RNA polymerase complex was determined using gel retardation. Alkylation was observed at all G residues in the template strand. Exposure of the alkylated DNA to Escherichia coli RNA polymerase resulted in the formation of a DNA-enzyme complex that was more stable, prior to initiation, than the complex formed with nonalkylated DNA. The DNA-RNA polymerase complex formed with the alkylated DNA also demonstrated decreased ability to progress along the full length of the DNA template. These observations show that, in addition to inducing transcriptional blockages, mustards also influence the interaction between RNA polymerase and its promoter. The ability to interfere with protein-DNA interactions may contribute significantly to the effects of these compounds in eukaryotic systems with their complex array of transcription factors. PMID- 8241139 TI - Interactions of bacteriophage T7 DNA primase/helicase protein with single stranded and double-stranded DNAs. AB - Protein-DNA interactions of bacteriophage T7 DNA primase/helicase protein 4A' with small synthetic oligodeoxynucleotides were investigated using a 20-base paired hairpin duplex, and 10-, 30-, and 60-base-long single-stranded DNA. The effect of nucleotide cofactors on DNA binding was examined using membrane binding assays which showed that 4A' binds DNA optimally only in the presence of MgdTMP PCP, the nonhydrolyzable analog of dTTP. About 20% of single-stranded DNA binding was observed in the presence of MgdTDP, but none was detectable in the absence of nucleotides. Native polyacrylamide gel electrophoresis showed that the DNAs bind predominantly to the hexameric form of 4A'. Larger oligomers of 4A' can bind DNA, but no DNA binding was observed to species smaller than the hexamer. Quantitative equilibrium binding studies at increasing 4A' concentrations and at increasing DNA concentrations showed tight binding of one 10-mer or 30-mer per hexamer. The 4A' hexamer can bind a second strand of DNA, but with a 50-fold weaker affinity than the first strand. The 60-mer showed tight binding to two 4A' hexamers, suggesting that a hexamer may interact with only 30-40 bases of single-stranded DNA. This was corroborated by nuclease protection experiments where the smallest length of DNA protected by 4A' or 4B protein was found to be about 30 bases. Equilibrium binding studies and competitive DNA binding data are consistent with a weaker affinity of 4A' for the duplex DNA. Only 20-25% of duplex DNA binding was observed at increasing 4A' protein in the presence of MgdTMP-PCP. About four duplex DNAs can bind each 4A' hexamer at increasing DNA concentrations, but their weaker binding was evident from their facile dissociation from 4A' in the presence of competing single-stranded DNA. PMID- 8241140 TI - Solution conformation of the (+)-trans-anti-[BPh]dA adduct opposite dT in a DNA duplex: intercalation of the covalently attached benzo[c]phenanthrene to the 5' side of the adduct site without disruption of the modified base pair. AB - Benzo[c]phenanthrene diol epoxide can covalently bind to the exocyclic amino group of deoxyadenosine to generate [BPh]dA adducts where the polycyclic aromatic hydrocarbon is attached to the major groove edge of DNA. This paper reports on NMR-energy minimization structural studies of the (+)-trans-anti-[BPh]dA adduct positioned opposite dT in the sequence context d(C5-[BPh]A6-C7).d-(G16-T17-G18) at the 11-mer duplex level. The exchangeable and nonexchangeable protons of the benzo[c]phenanthrenyl moiety and the nucleic acid were assigned following analysis of two-dimensional NMR data sets in H2O and D2O solution. The solution structure of the (+)-trans-anti-[BPh]dA.dT 11-mer duplex has been determined by incorporating intramolecular and intermolecular proton-proton distances defined by upper and lower bounds deduced from NOESY data sets as restraints in energy minimization computations. The covalently attached benzo[c]phenanthrene ring intercalates to the 5'-side of the [BPh]-dA6 lesion site without disruption of the flanking Watson-Crick dC5.dG18 and [BPh]dA6.dT17 base pairs. The observed buckling of the intercalation cavity reflects the selective overlap of the intercalated phenanthrenyl ring with dT17 and dG18 bases on the unmodified strand. The structure provides new insights into how a polycyclic aromatic hydrocarbon covalently attached to the major groove edge of deoxyadenosine can still unidirectionally intercalate into the helix without disruption of the modified base pair. Our study establishes that among the contributing factors are a propeller-twisted [BPh]dA6.dT17 base pair, displacement of the carcinogen-DNA linkage bond from the plane of the dA6 base, the specific pucker adopted by the benzylic ring, and the propeller-like nonplanar geometry for the aromatic phenanthrenyl ring system. Our combined experimental-computational studies to date have now identified three structural motifs adopted by covalent polycyclic aromatic hydrocarbon-DNA adducts with their distribution determined by the chiral characteristics of individual stereoisomers and by whether the covalent adducts are generated at the minor or the major groove edge of the helix. PMID- 8241141 TI - Structure of the protease from simian immunodeficiency virus: complex with an irreversible nonpeptide inhibitor. AB - A variant of the simian immunodeficiency virus protease (SIV PR), covalently bound to the inhibitor 1,2-epoxy-3-(p-nitrophenoxy)propane (EPNP), was crystallized. The structure of the inhibited complex was determined by X-ray crystallography to a resolution of 2.4 A and refined to an R factor of 19%. The variant, SIV PR S4H, was shown to diminish the rate of autolysis by at least 4 fold without affecting enzymatic parameters. The overall root mean square (rms) deviation of the alpha-carbons from the structure of HIV-1PR complexed with a peptidomimetic inhibitor (7HVP) was 1.16 A. The major differences are concentrated in three surface loops with rms differences between 1.2 and 2.1 A. For 60% of the molecule the rms deviation was only 0.6 A. The structure reveals one molecule of EPNP bound per protease dimer, a stoichiometry confirmed by mass spectral analysis. The epoxide moiety forms a covalent bond with either of the active site aspartic acids of the dimer, and the phenyl moiety occupies the P1 binding site. The EPNP nitro group interacts with Arg 8. This structure suggests a starting template for the design of nonpeptide-based irreversible inhibitors of the SIV and related HIV-1 and HIV-2 PRs. PMID- 8241142 TI - Mechanism of adenylate kinase. 1H, 13C, and 15N NMR assignments, secondary structures, and substrate binding sites. AB - Backbone 1H, 13C, and 15N NMR assignments were obtained for the complex of chicken muscle adenylate kinase (AK) with its bisubstrate analog, MgAP5A [magnesium P1,P5-bis(5'-adenosyl)-pentaphosphate]. The assignments were used to elucidate the secondary structures and the enzyme-MgAP5A interactions. The work involves two unusual features: the molecular weight of AK (21.6 kDa) is one of the largest, on a monomeric basis, for which nearly complete assignment has been reported to date, and the assignment was performed at pH 7.1 instead of the acidic pH used for most other proteins. The results are summarized as follows. Firstly, unambiguous sequential assignments of backbone resonances have been achieved effectively by the combined use of two sequential assignment methods: NOE-directed assignments and the recently developed 1J-coupling-directed assignments. The starting points of the assignments were provided by several specifically labeled enzyme samples. Over 90% of the backbone 1H, 13C, and 15N resonances have been assigned. Secondly, spin system information was obtained from the HCCH-TOCSY and HCCH-COSY experiments as well as from 2D homonuclear NMR data. Overall, the side-chain resonances of ca. 40% of the residues, including most of the those displaying NOEs with the adenosine moieties of MgAP5A, have been assigned. Thirdly, secondary structural elements in the AK-MgAP5A complex were identified by extensive analyses of 1H-15N 2D HMQC-NOESY and 3D NOESY-HMQC spectra. Overall, the enzyme consists of ca. 60% alpha-helices and a five stranded parallel beta-sheet. The results are compared with the secondary structure of the free AK from porcine muscle in crystals [Dreusicke, D., Karplus, P. A., & Schulz, G. E. (1988) J. Mol. Biol. 199, 359-371]. Lastly, most of the intermolecular NOEs between AK and the adenosine moieties of MgAP5A have been identified: Thr39, Leu43, Gly64, Leu66, Val67, Val72, and Gln101 are in proximity to the adenosine moiety of the adenosine 5'-monophosphate site, whereas Thr23 is in proximity to that of the adenosine 5'-triphosphate site. These data are discussed in relation to previous results from site-directed mutagenesis, NMR, and X-ray studies and in relation to the mechanism of catalysis. PMID- 8241143 TI - An NMR study of the HIV-1 TAR element hairpin. AB - The TAR hairpin is an important part of the 5' long terminal repeat of HIV-1 and appears to be recognized by a cellular protein. A 14-base model of the native TAR hairpin 5'-GAGC[CUGGGA]-GCUC-3' (loop bases in square brackets) has been studied by proton, phosphorus, and natural abundance carbon NMR; these results are compared to other published NMR studies of the TAR hairpin. Assignments of all nonexchangeable protons and of all the stem-exchangeable protons have been made, as well as all phosphorus and many carbon resonances. Large J1'2' and J3'4' proton-proton coupling in the C5, G8, and G9 sugars indicate an equilibrium between C2'- and C3'-endo forms; these data show a dynamic loop structure. We see three broad imino resonances that have not been reported before; these resonances are in the right region for unbonded loop imino protons. These peaks suggest the protons are protected from fast exchange with the solvent by the structure of the hairpin loop. Simulated annealing and molecular dynamics with 148 distance constraints, 11 hydrogen bonds, and 84 torsion angle constraints showed a wide variety of structures. Certain trends are evident, such as continuation of the A form helix on the 3' side of the hairpin loop. The ensemble of calculated structures agree with most chemical modification data. PMID- 8241144 TI - Pressure dependence of the melting temperature of dA.dT polymers. AB - The pressure dependence of the helix-to-coil transition temperature (Tm) of poly[d(A-T)], poly(dA).poly(dT), and poly(dA).poly(dT)2 was studied as a function of sodium chloride concentration. The molar volume change of the transition (delta V) was calculated using the Clapeyron equation and calorimetrically determined enthalpies. The delta V of the transition varied linearly with the logarithm of the salt concentration for both double-stranded polymers. The delta V of poly[d(A-T)] changed from +0.36 to +3.86 cm3 mol-1 as the sodium chloride concentration changed from 0.020 to 1.0 M. For poly(dA).poly-(dT), the delta V varied from +2.60 to +4.59 cm3 mol-1 over the range of 0.020-0.20 M NaCl. No pressure dependence of the van't Hoff enthalpy was observed for the double helical form of either polymer. The delta V for denaturation of the triple helical species poly(dA).poly(dT)2 was found to be +7.81 and +10.4 cm3 mol-1 at 1.0 and 3.0 M NaCl, respectively. The observed delta V values are interpreted in terms of changes in the hydration of the polymers arising from release of counterions and changes in the stacking of the bases upon denaturation. Counterion release is assumed to make a net negative contribution to the overall delta V, implying that disruption of the stacking interactions must make a positive volume change to the overall delta V. The difference in the delta V values for the two polymers remains constant as the salt concentration is changed, suggesting a difference in the partial molar volume of their single stranded forms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241145 TI - Characterization of the Pf3 single-strand DNA binding protein by circular dichroism spectroscopy. AB - We have used circular dichroism (CD) spectroscopy and gel electrophoresis to characterize the single-strand DNA binding protein (ssDBP) of the bacteriophage Pf3 and its complexes with Pf3 DNA and various DNA and RNA homopolymers. The secondary structure of Pf3 ssDBP had < 1% alpha-helix and therefore was probably a beta-sheet structure like the fd gene 5 protein (g5p). From CD titrations, the binding stoichiometry of Pf3 ssDBP was two nucleotides per protein monomer (n = 2) for complexes formed with all of the nucleic acids except poly[r(U)], for which n = 3 (in a buffer of 10 mM Tris-HCl and 70 mM NaCl, pH 8.2). Evidence of an additional binding mode of n = 4 for complexes formed with Pf3 DNA was found by gel electrophoresis experiments. Pf3 ssDBP showed a marked sequence dependence in binding affinities similar to that known for the fd g5p. PMID- 8241146 TI - Binding of heparin by type III domains and peptides from the carboxy terminal hep 2 region of fibronectin. AB - The major sites of heparin binding by fibronectin are located in fragments of 30 or 40 kDa that contain type III modules 12 through 14 or 15. Various proteolytic or recombinant subfragments and several synthetic peptides derived from this region have been compared with respect to their binding to fluorescein-labeled heparin in solution. Binding was monitored by the change in fluorescence anisotropy at 25 degrees C and pH 7.4 in 0.02 M Tris buffer, alone (TB) or with 0.15M NaCl (TBS). A 23-kDa fragment containing III13 and III14 but lacking III12 had Kd values of 0.3 and 1.8 microM in TB, and TBS, respectively, indistinguishable from the 30-kDa parent. Fragments containing only module III13 bound 2-3-fold weaker than the parent while those containing only III14 bound 6 50-fold weaker depending on the ionic strength. Fragments containing only III12 or III15 failed to bind at all in TBS. A cationic peptide derived from the amino terminus of III13 and containing the Arg-Arg-Ala-Arg consensus sequence, whose integrity was shown by Barkalow and Schwarzbauer [Barkalow, F. J., & Schwarzbauer, J. E. (1991) J. Biol. Chem. 266, 7812-7818] to be critical, failed to bind in TBS but bound weakly in TB. Two additional cationic peptides derived from the middle and C-terminal regions of III14 showed similar behavior. Thus while the major determinant(s) of heparin binding are located in III13, those determinants are only active when part of a properly folded structure. Furthermore, module III13 when isolated had a slightly lower affinity than fragments containing both III13 and III14.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241147 TI - Snapshots along the reaction coordinate of an SNAr reaction catalyzed by glutathione transferase. AB - The three-dimensional structures of a class mu glutathione transferase in complex with a transition-state analogue, 1-(S-glutathionyl)-2,4,6 trinitrocyclohexadienate, and a product, 1-(S-glutathionyl)-2,4-dinitrobenzene, of a nucleophilic aromatic substitution (SNAr) reaction have been determined at 1.9- and 2.0-A resolution, respectively. The two structures represent snapshots along the reaction coordinate for the enzyme-catalyzed reaction of glutathione with 1-chloro-2,4-dinitrobenzene and reveal specific interactions between the enzyme, intermediate, and product that are important in catalysis. The geometries of the intermediate and product are used to postulate reaction coordinate motion during catalysis. PMID- 8241148 TI - The active site structure of the calcium-containing quinoprotein methanol dehydrogenase. AB - Pyrroloquinoline quinone (PQQ), widely found in nature, serves as the redox cofactor in bacterial methanol dehydrogenase (MEDH), a heterotetrameric enzyme that oxidizes methanol to formaldehyde. The refined structure of MEDH at 2.4-A resolution, based on recently obtained amino acid sequence data, reveals that the PQQ, located in a central channel of the disk-shaped protein, is sandwiched between a Trp side chain and a very unusual vicinal disulfide. A Ca2+ ion forms a bridge between PQQ and the protein molecule, very close to a putative substrate binding pocket. The vicinal disulfide may form during PQQ incorporation and possibly act to hold the latter in place. PMID- 8241149 TI - A novel structural basis for membrane association of a protein: construction of a chimeric soluble mutant of (S)-mandelate dehydrogenase from Pseudomonas putida. AB - The (S)-mandelate dehydrogenase (MDH) from Pseudomonas putida (ATCC 12633) is the only membrane-associated member of a homologous family of FMN-dependent, alpha hydroxy acid dehydrogenases/oxidases that includes the structurally characterized glycolate oxidase from spinach (GOX). We have correlated the membrane association of MDH to a polypeptide segment in the interior of the primary sequence. This has been accomplished by construction of a chimeric enzyme in which the putative membrane-binding segment in MDH has been deleted and replaced with the corresponding segment from the soluble GOX. The resulting chimera, MDH-GOX, is soluble and retains partial catalytic activity (approximately 1%) using (S) mandelate as substrate. In contrast, the activities of both the membrane associated wild-type MDH and the soluble MDH-GOX are nearly the same when (S) phenyllactate is used as substrate. To the best of our knowledge, this is the first example of a membrane-associated protein in which an internal polypeptide segment anchors the protein to the membrane. PMID- 8241150 TI - Molecular cloning and chromosomal assignment of the gene for human Zn-alpha 2 glycoprotein. AB - Genomic clones containing the human Zn-alpha 2-glycoprotein gene were isolated. Two of them were for the functional gene overlapped, and the other four were for two different pseudogenes (1 and 2) retaining exon-intron organization. The complete DNA sequence of the functional gene (9.3 kb) and its 5'- and 3'-flanking regions (5.3 and 0.1 kb, respectively) was determined. The gene is composed of four exons; the first exon is for the 5'-untranslated region, the signal sequence, and the first six amino acids; the second, for domain A; the third, for domain B; and the fourth, for domain C and the 3'-untranslated region. The 5' flanking region contains a TATA box, a CAT box, an octamer sequence, and three possible Spl-binding sites. Ten and three copies of Alu repetitive DNA were identified within the gene and the 5'-flanking region, respectively, and they occupy 30% of the gene. The nucleotide sequences around the exons of pseudogene 1 were also determined; they had high homology (90-91%) with the corresponding region of the functional gene. Southern blot analysis suggested that there are only three genes, including nonfunctional ones, for Zn-alpha 2-glycoprotein in humans. The gene (ZA2G) was mapped to human chromosome band 7q22.1 by fluorescence in situ hybridization. PMID- 8241151 TI - Internal chemical bonding in solutions of simple phosphates and vanadates. AB - The chemical bonding within structurally related phosphates and vanadates in aqueous solution is compared on the basis of vibrational frequencies obtained by classical Raman spectroscopy. To do this, an empirical relationship between the stretching frequency of P-O and P-OH or P-OR groups and bond strength is developed such that the sum of the PO bond strengths, expressed in terms of average number of electron pairs per bond, is as close as possible to 5.0 for phosphoric acid and various anions and esters thereof. The same approach is used for the corresponding vanadates. The internal bonding in phosphates involves a greater bond strength for P-OH and a smaller strength for P-O than might be expected from a simple consideration of canonical resonance forms. In vanadates, V-OH and V-O are closer to single and double bonds, respectively, than in phosphates, and the force constant for V = O is considerably smaller than for P = O, although that for V-OH and P-OH is similar. Since treating the P-O and V-O groups of simple tetrahedral phosphates and vanadates as independent diatomic oscillators provides good correlations between the respective frequencies and bond strengths, the same correlations are used to approximate the expected stretching frequencies for distorted phosphates and vanadates. The distortions considered are those that presumably characterize associative and dissociative transition states for a concerted transfer of the (PO3-) fragment of a dianionic phosphate group between donor and acceptor oxygens with similar character.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241152 TI - Comparison of vibrational frequencies of critical bonds in ground-state complexes and in a vanadate-based transition-state analog complex of muscle phosphoglucomutase. Mechanistic implications. AB - The symmetric stretching frequency of the P-O bonds of the enzymic phosphate group in muscle phosphoglucomutase was measured via 16O/18O Raman difference spectroscopy. This frequency, and its shift on isotopic substitution, is characteristic of a dianionic phosphate ester. The P-O stretching frequency is not detectably altered by the binding of the metal ion activators Mg2+, Zn2+, or Cd2+ nor by the subsequent binding of glucose phosphate. Hence, a binding-induced distortion/polarization of the enzymic phosphate group in the ground state, or enzyme-substrate complex, cannot serve as a rationale for the large value of kcat in the phosphoglucomutase reaction. By contrast, the stretching frequency of the V-O bonds within a vanadate group bound at the same site in the transition-state analog complex involving glucose 1-phosphate 6-vanadate is much lower than for a normal dianionic vanadate. This low V-O stretching frequency is best rationalized in terms of the extensive polarization of all three nonbridging oxygens of the vanadate ester dianion plus the formation of a weak, fifth bond to the vanadium atom. This distortion/polarization of the VO3(2-) group depends on the metal ion activator, since it is largely abolished, and the involvement of the fifth ligand eliminated, by substitution of Li+ for Mg2+ at the metal activation site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241153 TI - Transition-state analysis of AMP deaminase. AB - The transition state of the allosteric AMP deaminase from Saccharomyces cerevisiae has been characterized by 14C and 15N Vmax/Km heavy-atom kinetic isotope effects. The primary 6-14C isotope effect was measured with [6-14C]AMP, and the 6-15N primary isotope effect was measured by isotope ratio mass spectrometry using the natural abundance of 15N in AMP and by using 15N release from ATP as a slow substrate. Isotope effects for AMP as the substrate were measured in the presence and absence of ATP as an allosteric activator and GTP as an allosteric inhibitor. Kinetic isotope effects with [6-14C]AMP were 1.030 +/- 0.003, 1.038 +/- 0.004, and 1.042 +/- 0.003 in the absence of effectors and in the presence of ATP and GTP, respectively. Isotope effects for [6-15N]AMP averaged 1.010 +/- 0.002. Allosteric activation increased the 15N isotope effect to 1.016 +/- 0.003. A primary 15N kinetic isotope effect with ATP, which has a Vmax/Km 10(-6) that for AMP, was 1.013 +/- 0.001. The presence of D2O as solvent caused a marginally significant decrease in the [6-15N]AMP kinetic isotope effect from 1.011 +/- 0.001 to 1.007 +/- 0.002. Previous studies have established that the solvent D2O effect is inverse (0.34) for slow substrates with two or more protons transferred prior to transition state formation and remains inverse (0.79) with AMP as substrate [Merkler, D. J., & Schramm, V. L. (1993) Biochemistry 32, 5792-5799]. Bond vibrational analysis was used to identify transition states for AMP deaminase that are consistent with all kinetic isotope effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241154 TI - Affinity labeling of glutathione S-transferase, isozyme 4-4, by 4 (fluorosulfonyl)benzoic acid reveals Tyr115 to be an important determinant of xenobiotic substrate specificity. AB - Incubation of 4-(fluorosulfonyl)benzoic acid (4-FSB), a xenobiotic substrate analogue, with the 4-4 isozyme of rat liver glutathione S-transferase at pH 7.5 and 25 degrees C results in a time-dependent inactivation of the enzyme. The rate of inactivation exhibits a nonlinear dependence on 4-FSB concentration from 0.50 to 7.85 mM, with kmax = 0.082 min-1 and a KI of 1.95 mM. Nearly 1 mol of reagent/mol of enzyme subunit is incorporated when the enzyme is maximally inactivated. Protection against incorporation and inactivation is provided by bromosulfophthalein, a competitive inhibitor with respect to the hydrophobic substrate, 1-chloro-2,4-dinitrobenzene (CDNB), suggesting that the reaction occurs in the binding site of the xenobiotic substrate. Fractionation by high performance liquid chromatography of a tryptic digest of inactivated enzyme yields a single, modified, 14-residue peptide containing Tyr115 as the altered amino acid. Modified and control enzymes have comparable affinities for glutathione, as indicated by fluorescence titration. In contrast, as distinguished from the control enzyme, modified enzyme does not adsorb to a column of an agarose-linked Cibacron Blue derivative, indicating that it has lost its ability to bind a hydrophobic substrate analogue. These results are supported by kinetic characteristics of modified and control enzymes: upon modification of the enzyme with 4-FSB, the apparent Km for glutathione is unchanged, while the apparent Km for CDNB increases dramatically from 193 to 1690 microM. When the reaction of 4-FSB with enzyme is monitored, the final percent residual activity is found to be dependent on the substrate used in the assay: 11% for CDNB, 20% for ethacrynic acid, 2.5% for trans-stilbene oxide, and 2% for trans-4-phenyl-3 butene-2-one. Analysis of the kinetics of modified enzyme suggests that Tyr115 of glutathione S-transferase, isozyme 4-4, contributes to xenobiotic substrate binding and, when certain types of substrates are employed, is involved in catalysis. PMID- 8241155 TI - Protein-ligand interactions in the lysine-binding site of plasminogen kringle 4 are different in crystal and solution. Electrostatic interactions studied by site directed mutagenesis exclude Lys35 as an important acceptor in solution. AB - Three amino acid residues previously reported to establish the interactions between lysine-like derivatives and plasminogen kringle 4 have been replaced by other residue types using the methods of site-directed mutagenesis. The effect of these modifications on the binding constant have been measured. The residues are Lys35, Asp57, and Arg71, according to the sequence numbering scheme adapted from the plasminogen kringle 5 domain. The plasminogen kringle 4 derivatives where Lys35 of the native molecule is replaced with isoleucine and methionine residues, respectively, were seen to bind the ligands, respectively, with association constants similar to those of the unmodified recombinant kringle 4 domain. The modification of Asp57 to asparagine was shown to eliminate the ability to bind to the lysine affinity column used to purify the protein. Similarly the site directed mutagenesis for Arg71 to glutamine resulted in a 12-19-fold decrease in binding of each of the two ligands. In addition, the effect of ionic strength on the binding of 6-aminohexanoic acid to the recombinant plasminogen kringle 4 and the three single substituted derivatives was examined. For the unmodified kringle domain as well as for the two derivatives modified only at the position of Lys35, an ionic strength of 0.5 M reduced the binding constant by a factor of 3 to 0.12 x 10(5) M-1. The derivative modified at the position of Arg71 was not effected by the ionic strength and maintained a rather low binding constant of 0.02 x 10(5) M 1. The observations suggest that the carboxylate of Asp57 and the guanidino group of Arg71 provide the electrostatic interaction in the binding site for the epsilon-amino group and the alpha-carboxylate of a C-terminal lysine residue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241156 TI - C-terminal peptide appendix in a class I tRNA synthetase needed for acceptor helix contacts and microhelix aminoacylation. AB - The 10 class I tRNA synthetases have an N-terminal nucleotide-binding fold which contains the catalytic center. Insertions into the nucleotide-binding fold provide contacts for acceptor-helix interactions, which stabilize the amino acid acceptor end of the tRNA substrate in the active site. A separate and largely nonconserved C-terminal domain provides contacts with distal parts of the tRNA, including the anticodon. For Escherichia coli methionyl tRNA synthetase, whose structure is known, the C-terminal domain is predominantly alpha-helical and forms a loop which interacts with the anticodon trinucleotide located about 76 A from the amino acid attachment site. Fused to the end of this helical domain is a peptide which curls back into the N-terminal nucleotide-binding fold and region of the active site. We show here that mutations in this peptide appendix disrupt aminoacylation and binding of a 7 base pair microhelix substrate based on the acceptor stem of tRNA(fMet), without affecting interactions with ATP or methionine or with the tRNA(fMet) anticodon. The impairment of acceptor-helix interactions by mutation of the C-terminal peptide can offset favorable anticodon interactions and severely reduce aminoacylation of tRNA(fMet). Thus, in addition to, or as an alternative to, acceptor-helix-binding insertions into the N terminal nucleotide-binding fold, C-terminal peptide epitopes in some class I enzymes may provide a mechanism for facilitating RNA microhelix interactions with the catalytic site. PMID- 8241157 TI - Incision at DNA G.T mispairs by extracts of mammalian cells occurs preferentially at cytosine methylation sites and is not targeted by a separate G.T binding reaction. AB - We have investigated the specificities of G.T mismatch binding proteins and of G.T mismatch cleavage in extracts of mammalian cells. G.T mismatch-specific protein:DNA complex formation by cell extracts was independent of the local sequence context of the mismatch. Cell extracts performed similar levels of protein binding to DNA substrates in which a single G.T mispair was preceded by T, G, A, C, or 5-meC. In contrast, incision by extracts of the T-containing strand of a G.T mismatch exhibited a strong sequence specificity and efficient strand cleavage was only observed when the mismatched G was in a CpG sequence. Thus, oligonucleotides containing either CpgGpT or 5meCpGGpT were efficiently incised, but not those containing GpGCpT, ApGTpT, or TpGApT sequences. Cell lines made resistant to the alkylating agent N-methyl-N-nitrosourea have previously been found to be defective in a G.T mismatch binding reaction. The defect in binding by extracts prepared from these cells extended to G.T mismatches in several sequence contexts. The variant extracts nevertheless incised G.T mismatches normally suggesting that this particular binding activity is not required for incision. The data indicate that incision by this activity is targeted to the CpG sequences in which G.T mismatches are formed by the mutagenic deamination of DNA 5-methylcytosine. In this regard the repair pathway resembles the very short patch (vsp) repair pathway in Escherichia coli. PMID- 8241158 TI - Evidence for the role of solvated metal hydroxide in the hammerhead cleavage mechanism. AB - The log of the rate of the chemical step of hammerhead cleavage in Mg2+ increases linearly with pH between pH 5.7 and 8.9. A slope of approximately 1 indicates that a single deprotonation is required for cleavage. Hammerhead pH-rate profiles with Ca2+, Mn2+, Co2+, and Cd2+ correlate well with the pKa's of these ions in water. This relationship between the pKa's and the pH-rate profile suggests that a metal hydroxide bound to the hammerhead RNA acts as the base in the cleavage mechanism. PMID- 8241159 TI - Three-dimensional structure of a simian immunodeficiency virus protease/inhibitor complex. Implications for the design of human immunodeficiency virus type 1 and 2 protease inhibitors. AB - Simian immunodeficiency virus (SIV) proteins have considerable amino acid sequence homology to those from human immunodeficiency virus (HIV); thus monkeys are considered useful models for the preclinical evaluation of acquired immune deficiency syndrome (AIDS) therapeutics. We have crystallized and determined the three-dimensional structure of SIV protease bound to the hydroxyethylene isostere inhibitor SKF107457. Crystals of the complex were grown from 25-32% saturated sodium chloride, by the hanging drop method of vapor diffusion. They belong to the orthorhombic space group I222, with a = 46.3 A, b = 101.5 A, and c = 118.8 A. The structure has been determined at 2.5-A resolution by molecular replacement and refined to a crystallographic discrepancy factor, R (= sigma parallel Fo magnitude of - magnitude of Fc parallel/sigma magnitude of Fo magnitude of), of 0.189. The overall structure of the complex is very similar to previously reported structures of HIV-1 protease bound to inhibitors. The inhibitor is bound in a conformation that is almost identical to that found for the same inhibitor bound to HIV-1 protease, except for an overall translation of the inhibitor, varying along the backbone atoms from about 1.0 A at the termini to about 0.5 A around the scissile bond surrogate. The structures of the SIV and HIV-1 proteins vary significantly only in three surface loops composed of amino acids 15-20, 34 45, and 65-70. Superposition of the 1188 protein backbone atoms from the two structures gives an rms deviation of 1.0 A; this number is reduced to 0.6 A when atoms from the three surface loops are eliminated from the rms calculation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241160 TI - Distal pocket polarity in ligand binding to myoglobin: deoxy and carbonmonoxy forms of a threonine68(E11) mutant investigated by X-ray crystallography and infrared spectroscopy. AB - The crystal structures of the deoxy and carbonmonoxy forms of a distal pocket myoglobin mutant in which valine68(E11) is replaced by threonine have been solved to 2.1- and 2.2-A resolution, respectively. This substitution has been shown previously to cause large decreases in the rate of oxygen binding and to lower the equilibrium association constants for O2 and CO. The synchrotron Laue method was used for the rapid acquisition of X-ray diffraction data to overcome problems caused by the very rapid rate of autooxidation of the mutant protein. The refined deoxy structure shows that the noncoordinated water molecule in the distal pocket is in a position to form strong hydrogen bonds with both the N epsilon-H of the distal histidine64 and O gamma of threonine68 with no other unexpected alterations in the protein structure. In the carbonmonoxy form, the bound ligand is well-defined and inclined away from the two hydrogen-bonding groups, refining to a position in which the Fe-C-O angle is 162 degrees. This value is very close to that previously observed in recombinant wild-type and position-64 (E7) mutants of sperm whale myoglobin (160-170 degrees). The similarity of the CO conformations contrasts with the 150-fold range in equilibrium binding constants (KCO) among the distal pocket myoglobin mutants and indicates that CO affinities cannot be predicted from the coordination geometry of the bound ligand. Furthermore, a comparison of the infrared stretching frequencies of CO in wild type, valine64 and threonine68 single mutant, and valine64-threonine68 double mutant pig carbonmonoxymyoglobins shows a lack of correlation between KCO and vCO. These effects can be understood in terms of the stability of noncovalently bound water in deoxymyoglobin and electrostatic interactions between bound ligands and the distal pocket residues. PMID- 8241161 TI - Sequence-specific assignments of the backbone 1H, 13C, and 15N resonances of the MutT enzyme by heteronuclear multidimensional NMR. AB - The MutT protein, a 129-residue enzyme from Escherichia coli which prevents A.T- >C.G mutations, catalyzes the hydrolysis of nucleoside triphosphates (NTP) to nucleoside monophosphates (NMP) and pyrophosphate [Bhatnagar, S. K., Bullions, L. C., & Bessman, M. J. (1991) J. Biol. Chem. 266, 9050-9054], by a mechanism involving nucleophilic substitution at the rarely attacked beta-phosphorus of NTP [Weber, D. J., Bhatnagar, S. K., Bullions, L. C., Bessman, M. J., & Mildvan, A. S. (1992a) J. Biol. Chem. 267, 16939-16942]. The bacterial MutT gene was inserted into the plasmid pET-11b under control of the T7 promoter and overexpressed in minimal media, permitting labeling of MutT with 13C and/or 15N. The yield after purification of the soluble fraction was approximately 35 mg of homogeneous MutT/L with physical and enzymatic properties indistinguishable from those of the originally isolated enzyme. Essentially complete sequence-specific assignments of the backbone HN, N, C alpha, H alpha, and CO resonances of the free enzyme (1.5 mM) were made at pH 7.4 and 32 degrees C, by heteronuclear double- and triple resonance experiments using a modified Bruker AM 600 NMR spectrometer. Specifically, 1H[15N]HSQC, 1H[15N]TOCSY-HMQC, and 1H[15N]NOESY-HMQC experiments were done with uniformly 15N-labeled enzyme. A 1H[15N] HSQC experiment was done with selective [alpha-15N]Lys-labeled enzyme. Also HNCA, HN(CO)CA, HNCO, constant time 1H[13C]HSQC, HCACO, and HCA(CO)N experiments were done with uniformly 13C- and 15N-labeled enzyme. Sequence-specific assignments were initiated from HN and 15N chemical shifts of Gly residues and of selectively labeled Lys residues in 1H[15N]HSQC experiments. They were confirmed by C alpha chemical shifts of Ala residues uniquely identified by residual coupling to C beta resonances in constant time 1H[13C]HSQC experiments. The sequence-specific assignments proceeded bidirectionally, terminating at Pro residues and at residues with undetectable NH signals, and the segments were linked to complete the backbone assignments. The backbone assignments reported here have permitted the interpretation of NOEs in the elucidation of the solution secondary structure of MutT, and the C alpha and H alpha chemical shifts have provided an independent approach to identifying secondary structural elements and to define their extent [Weber, D. J., Abeygunawardana, C., Bessman, M. J., & Mildvan, A. S. (1993) Biochemistry (following paper in this issue)]. PMID- 8241162 TI - Secondary structure of the MutT enzyme as determined by NMR. AB - The MutT enzyme (129 amino acids) catalyzes the hydrolysis of nucleoside triphosphates (NTP) to nucleotides (NMP) and pyrophosphate by nucleophilic substitution at the rarely attacked beta-phosphorus of NTP [Weber, D. J., Bhatnagar, S. K., Bullions, L. L., Bessman, M. J., & Mildvan, A. S. (1992) J. Biol. Chem. 267, 16939-16942]. Backbone NMR assignments for the H alpha, 13C alpha, HN, 15N, and carbonyl 13C' resonances, based on heteronuclear methods have been reported for MutT [Abeygunawardana, C., Weber, D. J., Frick, D. N. Bessman, M. J., & Mildvan, A. S. (1993) Biochemistry (preceding paper in this issue)]. Here, we report the secondary structure of MutT in solution on the basis of these assignments, NOE data derived from 2D and 3D homonuclear and heteronuclear NMR spectra, and amide NH exchange data. Consistent with near neighbor NOEs, H alpha and C alpha chemical shifts, and amide exchange rates, MutT contains two alpha helices spanning residues 47-59 (helix 1) and residues 119-128 (helix 2), respectively. The helical content predicted from NMR (17.8 +/- 1.0%) is consistent with that predicted by circular dichroism spectroscopy (20.9 +/- 5.4%). A mixed parallel and antiparallel beta-sheet with five beta-strands (A-E) consists of residues A, 3-13; B, 18-24; C, 70-74; D, 79-87; and E, 102 106.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241163 TI - NMR evidence for multiple conformations in a highly helical model peptide. AB - A monomeric model peptide, acetyl-WEAQAREALAKEAAARA-amide, has been structurally characterized using the complementary techniques of 1H 2D NMR and circular dichroism. Temperature-dependent CD measurements are consistent with a two-state helix/coil transition model and indicate a 65% contribution of helical conformers at 5 degrees C. Homonuclear 2D NMR experiments allowed the assignment of all proton resonances. The analysis of NOE-type cross-relaxation data established a large number of specific short- and medium-range NOE connectivities throughout the peptide, confirming the highly helical character of the peptide. However, the observation of long-range NOEs between the methyl protons of leucine-9 and backbone and side-chain protons of amino acids located at the N-terminus, as well as other unusual NOEs, unambiguously reflects the existence of significantly populated nonhelical structured conformers, indicating a multiconformational equilibrium. Implications of these observations with regard to secondary structure quantitation and current method limitations are discussed. PMID- 8241164 TI - Secondary structure and zinc ligation of human recombinant short-form stromelysin by multidimensional heteronuclear NMR. AB - Stromelysin-1, a member of the matrix metalloendoprotease family, is a zinc protease involved in the degradation of connective tissue in the extracellular matrix. As a step toward determining the structure of this protein, multidimensional heteronuclear NMR experiments have been applied to an inhibited truncated form of human stromelysin-1. Extensive 1H, 13C, and 15N sequential assignments have been obtained with a combination of three- and four-dimensional experiments. On the basis of sequential and short-range NOEs and 13C alpha chemical shifts, two helices have been delineated, spanning residues Asp-111 to Val-127 and Leu-195 to Ser-206. A third helix spanning residues Asp-238 to Gly 247 is characterized by sequential NOEs and 13C alpha chemical shifts, but not short-range NOEs. The lack of the latter NOEs suggests that this helix is either distorted or mobile. Similarly, sequential and interstrand NOEs and 13C alpha chemical shifts characterize a four-stranded beta-sheet with three parallel strands (Arg-100 to Ile-101, Ile-142 to Ala-147, Asp-177 to Asp-181) and one antiparallel strand (Ala-165 to Tyr-168). Two zinc sites have been identified in stromelysin [Salowe et al. (1992) Biochemistry 31, 4535-4540]. The NMR spectral properties, including chemical shift, pH dependence, and proton coupling of the imidazole nitrogens of six histidine residues (151, 166, 179, 201, 205, and 211), invariant in the matrix metalloendoprotease family, suggest that these residues are zinc ligands. NOE data indicate that these histidines form two clusters: one ligates the catalytic zinc (His-201, -205, and -211), and the other ligates a structural zinc (His-151, -166, and -179). Heteronuclear multiple quantum correlated spectra and specific labeling experiments indicate His-151, -179, 201, -205, and -211 are in the N delta 1H tautomer and His-166 is in the N epsilon 2H tautomer. PMID- 8241165 TI - Assignments for the main-chain nuclear magnetic resonances and delineation of the secondary structure of the catalytic domain of human stromelysin-1 as obtained from triple-resonance 3D NMR experiments. AB - We report the NMR assignments for the main-chain 13C, 15N, and 1H resonances (1HN, 1H alpha, 15N alpha, 13C alpha, 13CO) for the 19.5-kDa catalytic domain of human stromelysin-1, a zinc endoproteinase thought to be involved in pathologic tissue degradation. The assignments were predominantly obtained from triple resonance three-dimensional NMR experiments using double-labeled (15N/13C) samples. The secondary structure of the molecule was determined from analysis of 3D 15N-resolved NOESY experiments. It was found to consist of a five-stranded mixed beta-sheet with four parallel and one antiparallel strand and three helices. The topological arrangement of the secondary structure elements of stromelysin catalytic domain is remarkably similar to that found for astacin, a Zn proteinase for which the tertiary structure was recently determined from X-ray diffraction data [Bode et al. (1992) Nature 358, 164-167]. PMID- 8241166 TI - Sequential assignment and structure determination of spider toxin omega-Aga-IVB. AB - The solution structure of a peptide toxin isolated from funnel web spider venom, omega-Aga-IVB, was determined by 2D NMR methods. omega-Aga-IVB is a high-affinity specific blocker of P-type voltage-dependent calcium channels. Nearly all of the proton resonances of this 48-residue protein were assigned using conventional 2D homonuclear NMR experiments. The three-dimensional structure of the molecule was determined by simulated annealing. The distance and dihedral restraints used in the structure calculations were derived from NOESY and COSY-type experiments, respectively. Mass spectrometric analysis of omega-Aga-IVB suggests that the protein contains four disulfide bonds. In the absence of chemical data to identify the pattern of cysteine pairing, the disulfide bonds of the toxin are proposed from the NMR data and subsequent structural calculations. The structure of the toxin can be described as a three-stranded anti-parallel beta sheet connected by flexible loops. A striking feature of the structure is that the C terminal 10 residues of this protein adopt random coil conformations. Several positively charged amino acid side chains are found localized on one face of the molecule, in close proximity to the C-terminal tail. This observation has led us to propose a speculative model of the toxins blockade mechanism. PMID- 8241167 TI - Selective localization and rotational immobilization of univalent cations on quadruplex DNA. AB - The quadruplex structure of the oligomer d(T2G4T) is more stable in the presence of K+ than in the presence of Na+. This enhanced stability correlates with the preferential binding of K+ to a small number of specific sites on the quadruplex. In contrast, Na+ and K+ compete on an equal footing for atmospheric binding. Both 39K+ and 23Na+ are, when specifically bound, significantly inhibited in their rotational mobility, so that the quadrupolar relaxation reflects the molecular tumbling of the oligomer, which occurs on the time scale of nanoseconds. This rotational immobilization is in distinct contrast to the high rotational mobility of atmospherically bound cations. On the other hand, all NMR-visible 39K+ in solution is in rapid exchange among all environments (free, specifically bound, and atmospherically bound) implying that the lifetime of specifically coordinated 39K+ must be significantly shorter than a millisecond. A similar conclusion holds for 23Na+. The oligomer d(T2G4T) forms two distinct Hoogsteen base-paired structures in NaCl solution, separated by a large kinetic barrier. Neither of these structures is as stable with respect to base pair opening as is the quadruplex structure formed in KCl solution. Only one of these two structures is associated with rotational immobilization of bound 23Na+. PMID- 8241168 TI - Site-specific cleavage at a DNA bulge by neocarzinostatin chromophore via a novel mechanism. AB - The chromophore of the anticancer drug neocarzinostatin (NCS-Chrom) oxidatively cleaves single-stranded or duplex DNA site-specifically in the absence of activating thiol provided that the DNA contains a bulged structure. Point mutations, deletions, and insertions in the DNA analogue and its complement of the 3'-terminus of yeast tRNA(Phe) show that for a single-stranded DNA to be cleaved by NCS-Chrom the DNA must generate a hairpin structure with an apical loop and at least a two-base-pair stem hinged to a region of duplex structure via a bulge containing a target nucleotide at its 3' side. The size of the loop is not critical so long as it contains at least three nucleotides; the bulge requires a minimum of two nucleotides but must have fewer than five. With a notable exception involving base-pair changes immediately 3' to the bulge, base changes in the bulge and base-pair changes immediately 5' to the bulge retain substrate activity for NCS-Chrom. Maintenance of the bulged structure requires stable duplex regions on each side of the bulge. A similar bulged structure, but lacking a loop, formed by the annealing of a linear 8-mer and a 6-mer is an excellent target for cleavage in the thiol-independent reaction. Drugs such as netropsin, which sequester the DNA into nonbulge containing structures inhibit the reaction. In the absence of O2 strand cleavage is blocked and quantitatively replaced by a presumed drug-DNA covalent adduct.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241169 TI - Interactions of three strands in joints made by RecA protein. AB - RecA protein from Escherichia coli has been used to form a triple-stranded DNA structure from either single-stranded M13 DNA or a single-stranded oligonucleotide plus a duplex oligonucleotide with a hairpin loop. The secondary structure of purified deproteinized triplex was examined by probing with DNase I, P1 nuclease, potassium permanganate, and diethyl pyrocarbonate. The two strands destined to form heteroduplex DNA showed the same patterns of chemical modification and enzymatic digestion as control duplex DNA, indicating that they formed a normal duplex substructure. However, the nascent outgoing strand showed properties consistent with a novel triplex structure: most of its purine residues, especially adenines, were hyperreactive to all probes. The patterns of digestion by DNase I and P1 nuclease indicated that the nascent outgoing strand was not a freely mobile or single-stranded branch but rather was still interacting with the newly formed heteroduplex DNA. On the basis of the planar base triads proposed previously (Rao et al., 1993) and energy minimization of a third strand in the major groove of B-form DNA, we derived a model that helps to rationalize the properties revealed by chemical and enzymatic probing. PMID- 8241170 TI - Activation of the leu-500 promoter: a topological domain generated by divergent transcription in a plasmid. AB - The Salmonella typhimurium leu-500 promoter is active only in topA strains. In an earlier study (Chen, D., Bowater, R., Dorman, C., & Lilley, D. M. J. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 8784-8788), we showed that the activity of this promoter on a circular plasmid is a function of the transcription and translation of an adjacent tetA gene, and we suggested that the effect arises because of increased local negative superhelix density due to transcription (Liu, L. F., & Wang, J. C. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 7024-7027) initiated at the tetA promoter. In this study we show that translation of the 5' (N-terminal) section of tetA is required for activity of the leu-500 promoter, consistent with a requirement for membrane association of TetA. We have also shown the importance of a second transcription unit, the ampicillin resistance gene bla, in the activation of the leu-500 promoter. Thus the activity of the leu-500 promoter was reduced by partial deletion or premature termination of bla and was increased when the transcription of bla was boosted by the insertion of the stronger tac promoter. However, even in the latter situation the role of the tetA gene is dominant, and deletion of the tetA gene reduced activity of the leu-500 promoter to very low levels. These results suggest the existence of a topological domain defined by the divergent bla and tetA transcription units. Membrane insertion at tetA is essential to provide an anchorage point.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241171 TI - Equilibrium association constants for oligonucleotide-directed triple helix formation at single DNA sites: linkage to cation valence and concentration. AB - The linkage between the energetics of oligonucleotide-directed triple helix formation and the cationic solution environment has been investigated in mixed valence salt solutions. Equilibrium constants for formation of the local pyrimidine.purine.pyrimidine structure afforded by binding of the oligonucleotide 5'-d(T*TTTTCTCTCTCTCT)-3' to a single site within a 339-bp plasmid fragment were measured using quantitative affinity cleavage titrations at pH 7.0 and 22 degrees C in the presence of various concentrations of KCl, MgCl2, and spermine tetrahydrochloride (SpmCl4). In a solution containing 10 mM NaCl, 140 mM KCl, 1.0 mM MgCl2, and 1.0 mM SpmCl4, the measured binding constant was 3.3 (+/- 1.4) x 10(5) M-1. The equilibrium constant previously reported for the same association reaction in 100 mM NaCl and 1 mM SpmCl4 at the same temperature and pH was 10 fold higher [Singleton, S. F., & Dervan, P. B. (1992) J. Am. Chem. Soc. 114, 6957 6965]. Further study demonstrated that varying the potassium ion concentration between 5.0 and 140 mM (in the presence of 10 mM NaCl, 1.0 mM MgCl2, and 1.0 mM SpmCl4) resulted in an overall 100-fold decrease in the binding affinity from the lowest to the highest concentration. In contrast, measured binding constants increased 500-fold as the spermine concentration was increased from 0.40 to 4.0 mM (in the presence of 10 mM NaCl, 140 mM KCl, and 1.0 mM MgCl2). There was a modest effect on the binding constant (a 3-fold decrease) upon varying the magnesium ion concentration from 0.10 to 10 mM (in the presence of 10 mM NaCl, 140 mM KCl, and 1.0 mM SpmCl4).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241172 TI - Comparison of the membrane-bound states of two structurally similar delta selective opioid peptides by transferred nuclear Overhauser effect spectroscopy and molecular modeling. AB - NMR spectroscopic, peptide-membrane conformational studies on [D-Pen2,D-Pen5] enkephalin (DPDPE), an opioid receptor selective peptide, and an acyclic analog of DPDPE (DPDPE reduced at the disulfide bond) were conducted. The NMR method of transferred nuclear Overhauser effect (TRNOE) was used to obtain NOE profiles of the free and membrane bound forms of DPDPE and acyclic DPDPE. After comparison of the profiles of both peptides in the free and membrane-bound states, we hypothesize that the cyclic DPDPE undergoes little if any conformational change upon interaction with the membrane. However, for the acyclic analog, large changes in the NOE profile associated with backbone and side-chain groups were observed after interaction with the membrane. Results of computerized molecular modeling studies also were consistent with our theory that the free and membrane bound forms of cyclic DPDPE have very similar free and membrane-bound states. The free acyclic DPDPE has a reverse turn conformation with sidechains situated so that hydrophobic surface exposure to aqueous solution is minimized. After membrane interaction, the acyclic DPDPE has an extended conformation near the carboxy terminus with aromatic sidechains widely separated. We propose that the interaction of the acyclic DPDPE with the membrane surface is mediated by the amino terminus. We further propose that the interaction of the cyclic DPDPE with the membrane surface is limited because the D-Pen2 side chain is covalently bonded and the aromatic side chains and backbone are only slightly altered after membrane contact. Permeability studies by Ramaswami et al. [(1992) Biochim. Biophys. Acta 1109(2), 195-202] demonstrated that the acyclic DPDPE permeated through membranes at a rate 4 times greater than cyclic DPDPE. We conclude that conformational and topographical flexibility may be critical factors in peptide membrane interactions and permeability of bilayer membranes to opioid peptides. PMID- 8241173 TI - SecA, the peripheral subunit of the Escherichia coli precursor protein translocase, is functional as a dimer. AB - SecA, the peripheral ATPase domain of the Escherichia coli precursor protein translocase, was denatured in 6 M guanidine hydrochloride. Circular dichroism and intrinsic tryptophan fluorescence spectra revealed that the protein is transformed into a random-coil configuration. Upon dilution of the chaotropic agent, SecA refolds into its native, functional conformation as a homodimer. As structural criteria, the native dimeric state was assayed by size-exclusion chromatography, chemical cross-linking, tryptophan fluorescence, and circular dichroism. Functional SecA heterodimers were formed of which the individual subunits were tagged with fluorescent dyes to allow measurements of the association state of the monomers by resonance energy transfer using steady-state and time-resolved fluorescence spectroscopy. SecA retained its dimeric structure during translocation, while energy transfer was abolished only by denaturation. The "half-of-the-sites activity" was investigated by constructing heterodimers formed from native and 8-azido-ATP-inactivated SecA. Heterodimers have lost the ability to support translocation of the precursor protein proOmpA in an in vitro translocation system. It is concluded that the dimeric structure is maintained during translocation and required for functionality. PMID- 8241174 TI - Unfolding of the molten globule state of alpha-lactalbumin studied by 1H NMR. AB - The urea-induced unfolding of the molten globule state of bovine alpha lactalbumin was investigated by 1H nuclear magnetic resonance. In the molten globule state, most of the aromatic resonances deviate from their random coil values, indicating that aromatic side chains form some ordered structures in the molten globule state. When the urea concentration increases, the resonances are shifted, and the deviations from the random coil values are diminished. Because the chemical shifts of several random coil peptides are found to be independent of urea concentration, the urea-induced shifts of the resonances in the molten globule state reflect the unfolding transition of some ordered structures. The unfolding transitions measured by individual aromatic resonances do not coincide with each other. The unfolding transition curves obtained from some aromatic resonances are also different from those of the secondary structures measured by circular dichroism spectra. These results clearly show that the unfolding of the molten globule state of alpha-lactalbumin is not a cooperative two-state process. PMID- 8241175 TI - The effects of sulfite or nitrate on turnover-dependent inhibition in the ATPase from Halobacterium saccharovorum are related to the binding of the second metal ion. AB - The turnover-dependent inhibition of the Halobacterium saccharovorum ATPase is dependent on two parameters: pH and the concentration of the divalent cation present. At pH 6 and 1 mM Mn2+ the inhibition is small, but increases steeply with 6 mM Mn2+. In contrast, at pH 8.5 the inhibition is more than 90% at 1 mM Mn2+, and higher concentrations have little additional effect. A relationship between the occupation of a second metal ion binding site and turnover-dependent inhibition was postulated previously [Schobert, B. (1992) J. Biol. Chem. 267, 10252-10257]. The results lead to a model where this site (X-) can alternatively bind protons (XH), depending on the pH and the free metal ion concentration. The pKa of XH is estimated to be 9. The turnover-dependent inhibition is diminished by bisulfite, whereas sulfite is ineffective. The kinetics show that bisulfite and metal ion compete for the same site. In the proposed model, bisulfite binds via its negative charge to the site from which Pi was released and is arranged such as to interact with X- via its protonated group (X-HSO3-). In this way, formation of the inhibited enzyme species XMe is prevented. Inhibitory anions like nitrate, which carry a permanent dipole as a common feature, show uncompetitive inhibition vs metal ions. The data are compatible with a model in which these inhibitors bind to the vacant Pi site and position their positive charges near XH. As a consequence, the pKa of XH is decreased and X- is stabilized, which in turn favors the formation of XMe. The downshift in pKa was calculated to be 0.7 pH unit. PMID- 8241176 TI - Purine nucleoside phosphorylase. Catalytic mechanism and transition-state analysis of the arsenolysis reaction. AB - Purine nucleoside phosphorylase from calf spleen catalyzes the arsenolysis of inosine to form hypoxanthine and ribose 1-arsenate, which spontaneously hydrolyzes to ribose and arsenate. In the presence of H2(18)O, no 18O is incorporated into ribose, demonstrating that ribose 1-arsenate hydrolysis occurs by attack of water on the arsenic atom. Rapid reaction kinetics at 20 degrees C result in a biphasic rate curve with the first turnover occurring at a rate of 20 s-1 followed by a steady-state rate of 2 s-1. The product burst is consistent with rapid steps for substrate binding and arsenolysis followed by rate-limiting hypoxanthine release at a rate of 2 s-1. Purine nucleoside phosphorylase with bound [14C]inosine was mixed with excess unlabeled inosine and arsenate to determine relative rates for reaction or dissociation of bound inosine. The commitment factor (product formed/inosine released) was 0.19 at saturating arsenate, indicating that inosine binds to free enzyme and that bound inosine is not in thermodynamic equilibrium with free substrate. At neutral pH, kinetic isotope effects for the phosphorolysis reaction are small, indicating kinetic suppression. Kinetic isotope effects for arsenolysis were measured with [1'-3H]-, [2'-3H]-, [1'-14C]-, [9-15N]-, [4'-3H]-, and [5'-3H]inosine to provide experimental values of 1.118 +/- 0.003, 1.128 +/- 0.003, 1.022 +/- 0.005, 1.009 +/- 0.004, 1.007 +/- 0.003 and 1.028 +/- 0.004 respectively. Following correction for commitment factors, the intrinsic isotope effects were matched to a geometric transition-state model selected by bond-energy bond order vibrational analysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241177 TI - The importance of binding energy in catalysis of hydride transfer by UDP galactose 4-epimerase: a 13C and 15N NMR and kinetic study. AB - UDP-galactose 4-epimerase contains NAD+ irreversibly but noncovalently bound to the active site. Uridine nucleotides bind to the substrate site and induce a protein conformational change that increases the chemical reactivity of NAD+ at the coenzyme site. Activation of NAD+ by uridine nucleotides perturbs the 15N and 13C NMR chemical shifts of selectively enriched NAD+ bound to the coenzyme site. The proton-decoupled 15N NMR signal for enzyme-bound [carboxamide-15N]NAD+ does not change upon addition of UDP, indicating that activation is not brought about by a change in the binding of the carboxamide group. The 15N NMR signal of enzyme bound [nicotinamide-1-15N]NAD+ is shifted upfield 3.0 ppm and the 13C NMR signal for [nicotinamide-4-13C]NAD+ is shifted downfield 3.4 ppm downfield by the binding of UDP at the substrate site. These changes are consistent with the induction of a distortion into the nicotinamide ring, in which positive charge is transferred from N-1 to C-4. The kinetic and thermodynamic effects of these perturbations are significant, as indicated by the nonenzymatic chemical reactivities of a series of N-alkyl nicotinamides differing in the inductive electron withdrawing effects of the alkyl substituents. A downfield change of 3.4 ppm in the 4-13C chemical shifts brought about by electron withdrawal in the model compounds corresponds to a 3200-fold increase in the rate of reduction by NaBH3CN in water, a 15,000-fold increase in 86% ethanol, and a 152 mV more positive reduction potential in this series. The distortion of NAD+ by the binding of UDP is a long-range effect that is transmitted from the substrate binding site to the coenzyme through the protein conformational change. This apparently distorts the pi-electron distribution in the nicotinamide ring and reduces the activation energy for its reduction. Activation of enzyme-bound NAD+ toward reduction apparently arises from a destabilization in the nicotinamide ring structure rather than from a stabilization of the transition state through attractive interactions between the nicotinamide ring and the enzyme. PMID- 8241178 TI - Identification of lysine 153 as a functionally important residue in UDP-galactose 4-epimerase from Escherichia coli. AB - The role of lysine 153 in the action of UDP-galactose 4-epimerase from Escherichia coli has been investigated by site specific mutagenesis and kinetic and spectrophotometric analysis of the mutant enzymes. The crystal structure of UDP-galactose 4-epimerase shows that the binding of NAD+ to the coenzyme site includes the hydrogen bonded interaction of the epsilon-ammonium group of lysine 153 with the 2'- and 3'-hydroxyl groups of the nicotinamide riboside. Mutation of this residue to methionine or alanine decreases the catalytic activity of the enzyme by a factor of more than 10(3). The NAD+ associated with the wild type enzyme is subject to UMP-dependent reduction by sugars such as glucose and arabinose, but the mutant proteins K153M and K153A are not reduced by sugars in the presence or absence of UMP. NAD+ associated with the wild type enzyme is also subject to UMP-dependent reduction by sodium cyanoborohydride. However, although the mutant proteins bind UMP very well, the rate at which NAD+ associated with them is reduced by sodium cyanoborohydride is almost insensitive to the presence of UMP. The purified wild type enzyme contains significant amounts of NADH bound to the coenzyme site; however, the purified mutants K153M and K153A contain very little NADH. We conclude that lysine 153 plays an important role in increasing the chemical reactivity of enzyme-bound NAD+ in the uridine nucleotide-dependent conformational change associated with reductive inactivation and the catalytic activity of UDP-galactose 4-epimerase. PMID- 8241179 TI - Site-directed mutagenesis of tyrosine residues at nicotinamide nucleotide binding sites of Escherichia coli transhydrogenase. AB - Nicotinamide nucleotide transhydrogenase (E.C.1.6.1.1) from Escherichia coli was investigated with respect to the role of specific conserved tyrosine residues of putative substrate-binding regions. The enzyme from E. coli is made up of two subunits, alpha (510 residues) and beta (462 residues). The corresponding enzyme from bovine mitochondria is a single polypeptide (1043 residues) whose N-terminal region corresponds to the alpha subunit and whose C-terminal region corresponds to the beta subunit. Tyrosines 245 and 1006 of the mitochondrial enzyme have been shown to react selectively with 5'-(p-fluorosulfonylbenzoyl)adenosine with inactivation of the enzyme. In E. coli these residues correspond to tyrosine 226 of the alpha subunit and tyrosine 431 of the beta subunit. In addition, tyrosine 315 of the beta subunit is of interest since mutation of an adjacent residue (glycine 314) leads to inactivation [Ahmad, S., Glavas, N. A., & Bragg, P. D. (1992) Eur. J. Biochem. 207, 733-739]. In order to assess the role of the aforementioned conserved tyrosine residues in the mechanism and structure of transhydrogenases, these were replaced by site-specific mutagenesis, using the cloned and overexpressed E. coli transhydrogenase genes [Clarke, D. M., & Bragg, P. D. (1985) J. Bacteriol. 162, 367-373]. Phenylalanine mutants of all three tyrosine residues showed approximately 50% activity or more with regard to catalytic activity assayed as reduction of 3-acetylpyridine-NAD+ by NADPH. These mutants were also active in proton pumping assayed as quenching of 9-methoxy-6 chloro-2-aminoacridine or quinacrine fluorescence.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241180 TI - Electron transfer from cytochrome c2 to the primary donor of Rhodobacter sphaeroides reaction centers. A temperature dependence study. AB - Kinetics of flash-induced electron transfer from the soluble cytochrome c2 to the primary donor (P) of the reaction center purified from the purple bacterium Rhodobacter sphaeroides R-26 were investigated by time-resolved absorption spectroscopy. Re-reduction of P+ induced by a laser pulse was measured at 1283 nm both in isolated reaction centers and in reconstituted proteoliposomes reproducing the lipid composition of the native membrane. The effects of temperature (230-300 K) and of the cytochrome c2/reaction center stoichiometry were examined. At room temperature, over a wide range of cytochrome c2 to reaction center molar ratios, the biphasic kinetics of cytochrome c2 oxidation in the microsecond-to-millisecond time scale could be accurately described by a minimum reaction scheme which includes a second-order collisional process (k = 1.4 x 10(9) M-1 s-1 and k = 2.4 x 10(9) M-1 s-1 in isolated and reconstituted reaction centers, respectively) and a first-order intracomplex electron donation (t1/2 = 590 +/- 110 ns in isolated reaction centers; t1/2 = 930 +/- 140 ns in proteoliposomes). At cytochrome c2 to reaction center molar ratios exceeding 5, the monomolecular process almost completely accounts for P+ re-reduction. At lower stoichiometries, the relative contribution of the two parallel reaction pathways is modulated by a single binding equilibrium between cytochrome c2 and reaction centers, yielding a binding constant of 3.5 x 10(5) M-1 in both systems.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241181 TI - Spectroscopic characterization of mutants supports the assignment of histidine 419 as the axial ligand of heme o in the binuclear center of the cytochrome bo ubiquinol oxidase from Escherichia coli. AB - The bo-type ubiquinol oxidase of Escherichia coli is a member of the superfamily of heme-copper oxidases which also includes the aa3-type cytochrome c oxidases. The oxygen-binding binuclear center of cytochrome bo is located in subunit I and consists of a heme (heme o; heme a3 in the aa3-type oxidases) and a copper (Cu(B)). Previous spectroscopic studies have shown that heme o is bound to the protein via a single histidine residue. Site-directed mutagenesis of conserved histidine residues in subunit I has identified two residues (H284 and H419) which are candidates for the ligand of heme o, while spectroscopic studies of mutants at H284 definitively demonstrated that this residue cannot be the axial ligand. Consequently, the single remaining conserved histidine in subunit I (H419) was assigned as the ligand for the heme of the binuclear center. In this paper, this assignment is tested by characterization of additional mutants in which the putative heme o axial ligand, H419, is replaced by other amino acids. All mutations at H419 result in the loss of enzyme activity. Analyses via UV-visible and Fourier transform infrared spectroscopies reveal that substantial perturbation has occurred at the binuclear center as a result of the amino acid substitutions. In contrast with the wild-type enzyme, the mutant enzymes bind very little carbon monoxide. Three other amino acid residues which are potential ligands for heme o are shown tob e nonessential for enzyme activity. Mutations in these residues do not perturb the UV-visible or FTIR spectroscopic characteristics of the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241182 TI - Formation of iron(III)-tyrosinate is the fastest reaction observed in ferritin. AB - Rapid mineralization of ferritin, characteristic of protein with H-type subunits, coincides with formation of a specific Fe(III)-tyrosinate complex. The pseudo first-order rate constant for Fe(II) oxidation by H-subunit-type ferritin has now been shown to be 700-900 times greater than any previously reported for ferritin; kox = 1000 s-1 for formation of the specific Fe(III)-tyrosinate complex (A550nm) or formation of less defined Fe(III)-oxo multinuclear complexes (A420nm). Formation of multinuclear Fe(III)-oxo complexes and O2 consumption were biphasic. In the first phase, up to 50 Fe atoms/ferritin molecule were rapidly oxidized, accompanied by formation of the Fe(III)-tyrosinate complex; saturation of the sites which formed the Fe(III)-tyrosinate complex also required 50 Fe/ferritin molecule. The sigmoidal shape of the curve obtained by plotting the initial rate of oxidation during the rapid phase of mineralization versus added [Fe(II)] suggested a more complex reaction pathway of ferroxidation than previously described. During the second phase of mineralization, Fe(III)-tyrosinate decreased, but multinuclear Fe(III)-oxo complexes and O2 consumption continued to increase at a slower rate. Recovery of the rapid oxidation pathway paralleled recovery of the site for Fe(III)-tyrosinate formation; full regeneration of the Fe(III)-tyrosinate sites was gradual over a period of 12 h, as if the movement of Fe(III) along the biomineralization pathway in the protein was slow and was accompanied by conformational changes which affected the Fe(III)-tyrosinate site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241183 TI - Detection of bovine heart mitochondrial cytochrome c oxidase dimers in Triton X 100 and phospholipid vesicles by chemical cross-linking. AB - Bovine heart cytochrome c oxidase is a multisubunit enzyme whose oligomeric state is dependent on its detergent or phospholipid environment. We have utilized the cleavable, heterobifunctional cross-linking reagent N-succinimidyl 3-[(4 azidophenyl)dithio]propionate (SADP) to detect cytochrome c oxidase dimers. Monomeric or dimeric enzyme dispersed in Triton X-100 (as assessed by sedimentation velocity measurements) was reacted with SADP. A unique intersubunit cross-link having an apparent molecular mass of 136 kDa was identified in the dimeric enzyme; this product was insensitive to limited proteolysis by trypsin and contained a cross-link between two adjacent monomers. Two-dimensional NaDodSO4-PAGE (the second dimension containing beta-mercaptoethanol to cleave the cross-linking reagent) indicated that subunit I was the major component of the dimer-specific cross-link. The dimer-specific cross-link created by SADP was observed in phospholipid vesicles [cardiolipin/phosphatidylcholine (1:20, w/w)] containing dimeric (2 microM heme aa3) enzyme; a low yield of dimer-specific cross-link was observed in liposomes containing 6 microM (heme aa3) monomeric enzyme. The 136-kDa cross-link was not observed in liposomes containing 2 microM (heme aa3) monomeric enzyme. These results indicate that subunit I from each monomer may provide one site of interaction between monomers in the dimeric form of the enzyme and that cytochrome c oxidase monomers may reassociate to form dimeric complexes in phospholipid vesicles. PMID- 8241184 TI - A 13C and 2H nuclear magnetic resonance study of phosphatidylcholine/cholesterol interactions: characterization of liquid-gel phases. AB - A detailed study on the structure, dynamics, and thermodynamic behavior of phosphatidylcholine/cholesterol (PC/CHOL) mixtures was undertaken using differential scanning calorimetry (DSC) and solid-state nuclear magnetic resonance (NMR) spectroscopy. DSC thermograms of mixtures of cholesterol (CHOL) with 1,2-dipalmitoyl-sn-phosphatidylcholine (DPPC), 1,2-distearoyl-sn phosphatidylcholine (DSPC), and 1,2-diarachidoyl-sn-phosphatidylcholine (DAPC) showed a broadening of the first-order gel-->liquid crystalline transition and a decrease in the transition enthalpy, indicating a gradual loss of cooperativity for high CHOL concentrations. DPPC and DSPC were labeled with 13C at the carbonyl group of the sn-2 chain and 2H was introduced into the middle of the sn-2 chain at the 6- and 12-position for DPPC and DSPC, respectively. The 13C and 2H NMR spectra of each labeled lipid were studied as a function of temperature and CHOL concentration. The residual quadrupole splitting in the 2H NMR spectra, delta nu Q perpendicular, was analyzed as a function of temperature and composition. For CHOL concentrations less than 30 mol %, a precipitous change in delta nu Q perpendicular occurs near the chain melting temperature of the phospholipid. Further increases in CHOL concentration broaden the transition and shift the midpoint to higher temperature, indicating the presence of a new phase at higher CHOL contents. Moreover, at a given temperature, delta nu Q perpendicular increases with increasing cholesterol content, which indicates a more ordered structure. The 13C NMR spectra in the gel state consisted of a superposition of two components which can be attributed to both gel-like and fluid phospholipid domains in the bilayer. This two-component spectrum can be simulated quantitatively with a two-parameter chemical exchange model, which permits the fraction of each form and the exchange rate to be determined as a function of temperature and composition. At high CHOL contents the line width of the fluid component broadens, suggesting an increase in the exchange rate between the domains. These results were interpreted in terms of a temperature composition diagram with one region L beta', two regions LGI and LGII, and one liquid crystalline region L alpha, with LG denoting "liquid-gel" type phases. Liquid-gel phases correspond to phases with increased order in the hydrocarbon chains (in comparison to that of the pure PC bilayer in the L alpha phase) combined with fast limit axial diffusion that averages the 13C NMR spectrum to a "fluidlike" line.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8241185 TI - Use of fast protein size-exclusion liquid chromatography to study the unfolding of proteins which denature through the molten globule. AB - Fast protein size-exclusion liquid chromatography (SEC-FPLC) was used to study solvent-induced unfolding of six proteins. Two of them (sperm whale myoglobin and hen white lysozyme) denature on the simple N (native)<-->U (completely unfolded) scheme. The other four proteins [bovine and human alpha-lactalbumin, bovine carbonic anhydrase B (BCAB), and beta-lactamase from Staphylococcus aureus] denature through the molten globule (MG) state (i.e., on the N<-->MG<-->U denaturation scheme). We have shown that the permeation properties of the Superose 12 columns are practically independent of temperature, pH, and denaturants in wide concentration intervals. In the case of myoglobin and lysozyme denaturation at 4 degrees C (when the exchange between the native and unfolded states is slower than the characteristic time of chromatography), a bimodal distribution on molecular dimensions in the transition region was observed. This indicates that, under denaturant action, protein molecules can only be in one of the two states with different compactness. In other words, this shows that FPLC is one of the most direct approaches to establish the "all-or none" mechanism of the equilibrium solvent-induced denaturation of globular proteins. The curves of guanidinium hydrochloride- (GdmHCl) or urea-induced unfolding (N<-->U or MG<-->U transitions) of a protein on a column (monitored either by the relative areas of two peaks or--for fast exchange--by the position of the average peak) coincide with those monitored by far-UV CD in solution. The Stokes radius values obtained with the use of FPLC for the molten globule states of BCAB (1.6 M GdmHCl in 0.1 M sodium phosphate, pH 6.8, and acid form at pH 3.6) and for the human alpha-lactalbumin molten globule (2.0 M GdmHCl in 0.1 M sodium phosphate, pH 6.8) coincide with those known from literature. Thus, it has been shown that fast protein size-exclusion liquid chromatography (FPLC) is an "inert" technique, i.e., it does not shift the equilibrium between N, MG, and U states and, therefore, can be used for qualitative and quantitative studies of protein denaturation. PMID- 8241186 TI - Backbone dynamics of (Pro-Hyp-Gly)10 and a designed collagen-like triple-helical peptide by 15N NMR relaxation and hydrogen-exchange measurements. AB - The backbone dynamics of specific residues in two collagen-like triple-helical peptides with (X-Y-Gly)n sequences have been investigated using two-dimensional inverse-detected 15N NMR relaxation measurements and hydrogen-exchange experiments. One peptide, (POG)10, has the highest possible imino acid content and is considered to be a very stable prototype of a triple helix. The second peptide, (POG)3ITGARGLAGPOG(POG)3 (denoted T3-785), models an imino acid poor region of type III collagen and contains 12 residues from near the unique collagenase cleavage site. 15N relaxation parameters and hydrogen-exchange data were obtained for a glycine residue in the center of (POG)10 and for the tripeptide unit Gly-Leu-Ala in the middle of T3-785. Analysis of the relaxation data of the rodlike triple-helical peptides required the assumption of anisotropic overall motion, and the model-free approach of Lipari and Szabo (1982) was used to derive overall motional parameters and the order parameter, S2, that describes the amplitudes of the internal motion. First the mobilities of the Gly, Leu, and Ala residues in peptide T3-785 were compared. Both hydrogen exchange methods and relaxation measurements indicated that the residue in the Y position (Ala) is more mobile than residues in the Gly and X positions (Leu). The slower exchange rates of Gly and Leu compared to that of Ala are consistent with the two-hydrogen-bonded model for the triple helix. Then the backbone mobilities of the central Gly residue were compared for the two peptides (POG)10 and T3-785. In this case, 15N relaxation measurements give different results from hydrogen exchange. The glycine residues in the trimer form of both T3-785 and (POG)10 have high values for the order parameter (near 0.85), suggesting similar small amplitude internal motions and rigid backbones in both peptides. In contrast to the similar values of the order parameters, hydrogen-exchange data indicate that the central Gly exchanges at a faster rate in the trimer form of T3-785 than in (POG)10. These results suggest that a Gly in the imino acid rich environment of (POG)10 is dynamically different from a Gly in the imino acid poor environment of T3-785 and that the difference lies in the slower motion related to stability, rather than the faster motion on the picosecond time scale. This sequence dependent difference in dynamical properties may have important consequences for recognition processes in collagen. PMID- 8241187 TI - Quantitation of the interaction of protein kinase C with diacylglycerol and phosphoinositides by time-resolved detection of resonance energy transfer. AB - Quantitative studies of the binding of protein kinase C (PKC) to lipid cofactors were performed by monitoring resonance energy transfer with time-resolved fluorescence techniques. For that purpose, diacylglycerol (DG), phosphatidylinositol 4,5-biphosphate (PIP2), phosphatidylinositol 4-phosphate (PIP), phosphatidylinositol (PI), phosphatidylcholine (PC), and phosphatidylserine (PS) were labeled with a pyrenyl decanoyl moiety at the sn-2 position of the lipid glycerol. These labeled lipids proved excellent energy acceptors of light-excited tryptophan residues in PKC. The quenching efficiency of the tryptophan fluorescence was determined as function of lipid probe concentration in mixed micelles consisting of poly(oxyethylene)-9-lauryl ether, PS, and various mole fractions of probe lipid. The experimental conditions and method of data analysis allowed the estimation of binding constants of single or multiple pyrene lipids to PKC. The affinity of PKC for inositide lipids increases in the order PI < PIP < PIP2. The affinity of PKC for PIP and PIP2 is higher than that for DG. Determination of PKC activity in the presence of labeled lipids and PS showed that only PIP2 and DG activate PKC. Double-labeling experiments suggest that PIP2 and DG are not able to bind simultaneously to PKC, indicating a reciprocal binding relationship of both cofactors. The results support the notion that, besides DG, PIP2 can be a primary activator of PKC. PMID- 8241188 TI - Spontaneous interbilayer transfer of phospholipids: dependence on acyl chain composition. AB - The diffusion-mediated intervesicle transfer of a variety of fluorescent N-(S bimanylmercaptosuccinyl)phosphatidylethanolamines (BMS-PEs), and of a more limited series of radiolabeled phosphatidylcholines (PCs), has been investigated as a function of the acyl chain composition of the labeled molecules and the composition of the lipid vesicles. Increasing the total acyl chain length of a BMS-PE by two methylene residues increases the free energy of activation for probe desorption from a lipid bilayer (delta Gd not equal to) by ca. 1300 cal mol 1, while the addition of one double bond to either chain decreases delta Gd not equal to by a slightly smaller increment. The values of these increments in delta Gd not equal to are surprisingly consistent with variations in BMS-PE acyl chain length, positional isomerism (for heteroacyl BMS-PEs), and the sterol content or degree of acyl chain saturation of the lipid bilayer. However, the successive addition of multiple cis double bonds to the same acyl chain leads to progressively smaller decrements in the value of delta Gd not equal to. Radiolabeled PCs show somewhat lower absolute rates of interbilayer transfer than the corresponding BMS-PEs, but the incremental contributions of added methylene groups or cis double bonds to delta Gd not equal to are essentially identical for the two classes of lipids, validating the use of BMS-PEs as representative probes of phospholipid behavior for these experiments. Our data suggest that the relative rates of spontaneous transfer of a variety of phospholipids from bilayer membranes can be predicted to good approximation from a few basic features of the phospholipid structure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241189 TI - Calponin-calmodulin interaction: properties and effects on smooth and skeletal muscle actin binding and actomyosin ATPases. AB - Smooth muscle calponin bound to the biologically active fluorescent calmodulin [2 (4'-maleimidoanilino)naphthalene-6-sulfonic acid-calmodulin] (MIANS.CaM) with a Kd of 80 nM and produced a 3.4-fold fluorescence enhancement. PKC-phosphorylated calponin (1.3 mol of Pi/mol) bound to CaM with approximately 15-fold lower affinity. Calponin inhibited CaM (10 nM) activation of the Ca(2+)-/CaM-activated cyclic nucleotide phosphodiesterase (PDE) with an IC50 of 138 nM. The calponin CaM interaction was Ca(2+)-dependent: half-maximal binding of calponin to MIANS.CaM occurred at pCa 6.6 with a Hill coefficient of 2.4. Stopped-flow fluorescence kinetic analysis demonstrated that EGTA chelation of Ca2+ from CaM disrupted the MIANS.CaM-calponin complex at a rate of 1 s-1. Calponin bound MIANS.CaM at a rate of (6.0 +/- 1.8) x 10(6) M-1s-1, and melittin and unlabeled brain CaM both disrupted the MIANS.CaM-calponin complex at a rate of 0.3 +/- 0.1 s-1. These studies suggest that calponin binds CaM with 80-fold lower affinity than myosin light-chain kinase and that calponin associates with CaM much slower than it associates with caldesmon or myosin light-chain kinase. The physiological relevance of the CaM-calponin interaction was evaluated by analysis of the effects of Ca(2+)-CaM on (i) the interaction of calponin with actin and (ii) calponin-mediated inhibition of actin-activated myosin MgATPase activity. Ca(2+) CaM half-maximally inhibited calponin (2 microM) binding to smooth and skeletal muscle actins (9 microM) at 5.4 and 11 microM CaM, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241190 TI - Glutamate 329 located in the fourth transmembrane segment of the alpha-subunit of the rat kidney Na+,K+-ATPase is not an essential residue for active transport of sodium and potassium ions. AB - An allelic variant of the ouabain-insensitive rat kidney Na+,K(+)-ATPase alpha 1 isoform was identified by chance in a cDNA library. The variant differed from the wild-type rat kidney Na+,K(+)-ATPase by a single G to C base substitution in the cDNA, which on the amino acid level gave rise to a glutamine in place of the glutamate residue Glu329, previously suggested as a likely donator of oxygen ligands for Na+ and K+ binding. The variant cDNA was transfected into COS-1 cells and the transfectants expanded with success into stable cell lines that were able to grow in the presence of a concentration of ouabain highly cytotoxic to the parental cells containing only the endogenous COS-1 cell Na+,K(+)-ATPase. Under these conditions, the viability of the cells depended on the cation transport mediated by the ouabain-insensitive Glu329-->Gln variant, whose cDNA was shown by polymerase chain reaction amplification to be stably integrated into the COS-1 cell genome. Functional analysis on isolated plasma membranes demonstrated that the Glu329-->Gln variant was able to catalyze Na(+)- and K(+)-activated ATPase activity with a maximum turnover number similar if not identical to that of the wild type, but the variant exhibited a reduced affinity for both cations corresponding to a 2-fold increase in K0.5 for Na+ and a 6-fold increase in K0.5 for K+. Moreover, the apparent affinity for ATP was increased 15-fold in the variant relative to wild-type Na+,K(+)-ATPase. The Na+,K(+)-ATPase activity of the variant displayed an anomalous pH dependence with a down-shift of the pH optimum and a nearly constant rate in the range between pH 7.0 and 8.7. PMID- 8241191 TI - Heterodimers of the neurotrophic factors: formation, isolation, and differential stability. AB - We have determined that all four known members of the neurotrophin family, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT 3), and neurotrophin 4 (NT-4), are capable of forming noncovalent heterodimers. The formation of these heterodimers was accomplished by homodimer subunit exchange promoted by treatment with guanidine hydrochloride, urea, low pH, or acetonitrile. In some cases (BDNF and mouse NGF; BDNF and NT-4), generation of the heterodimers was achieved by incubating homodimer mixtures in a neutral buffer at ambient temperature. The formation of heterodimers was in each case detected by nondenaturing gel electrophoresis at pH 7.4. High-performance cation exchange chromatography was used to separate neurotrophin heterodimers from their parental homodimers. Heterodimers between BDNF and NT-3, BDNF and NT-4, and NT-3 and NT-4 are stable and show only a very small increase in homodimer content after 24 h of incubation at 37 degrees C. In contrast, heterodimers containing NGF subunits undergo gradual rearrangement to the homodimers. Our studies indicate that low pH, acetonitrile, and urea merely increase the neurotrophin subunit exchange rate and decrease the time needed to reach an equilibrium between a heterodimer and its two parental homodimers. PMID- 8241192 TI - Role of phosphate-magnesium-binding regions in the high GTPase activity of rac1 protein. AB - rac1, a member of the low molecular mass GTP-binding protein family, has a 20 fold higher GTPase activity than H-ras, but the structural motifs responsible for this property do not appear to reside within the conserved amino acids of the consensus GTP-binding domains [Menard, L., Tomhave, E., Casey, P.J., Uhing, R.J., Snyderman, R., & Disbury, J.R. (1992) Eur. J. Biochem 206, 537-546]. In this study, we determined the contribution, to the GTPase activity, of additional amino acids found in the phosphate-magnesium-binding (PM) regions of rac1. rac1 has three different amino acids from H-ras in each of the first two PM regions, while the third PM region is identical to that of H-ras. Mutation of the amino acids in the first PM region (aa 10-17) to the corresponding amino acids found in H-ras showed that modification of one of them, Asp11, resulted in a 50% decrease of the GTPase in rac1, whereas Gly13 and Ser17 had no effect. In the second PM region (aa 29-35), modification of the Pro29-Gly30 pair also reduced GTPase activity by 50% in rac1. rac1 mutated at positions 11 and 29 as well as 30 (P1-P2 mutant) had a 3-4-fold reduced GTPase activity compared to native rac1 (190 vs 552 pmol of GTP hydrolyzed/nmol of GTP gamma S-binding protein/min at 37 degrees C), suggesting a cooperative (but nonadditive) interaction between both domains. All mutants with reduced GTPase activity also had reduced affinity for GDP, shown by both equilibrium binding and GDP dissociation measurements.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241193 TI - Reaction of the ArsA adenosinetriphosphatase with 2-(4' maleimidoanilino)naphthalene-6-sulfonic acid. AB - The oxyanion-translocating ATPase encoded by the plasmid-borne ars operon catalyzes extrusion of antimonials and arsenicals from cells of Escherichia coli, thus providing resistance to those toxic oxyanions. The purified catalytic subunit of the ATPase, the ArsA protein, exhibits oxyanion-stimulated ATPase activity. The nature of the oxyanion binding site was probed by reaction with the fluorescent sulfhydryl probe 2-(4'-maleimidoanilino)naphthalene-6-sulfonic acid (MIANS). Our results indicate that MIANS reacts with the ArsA protein in an antimonite-dependent manner. After the protein had been modified with MIANS, two of four cysteines in the ArsA protein had reacted with the probe in the absence of the oxyanionic substrate, and three in the presence of antimonite. The quantum yield of the MIANS-ArsA protein adduct was significantly higher if modification of the protein had occurred in the presence of oxyanionic substrates. Thus binding of the anionic substrate of the pump produces a conformational change in the ArsA protein such that a single additional cysteinyl residue reacts more readily with the sulfhydryl probe. PMID- 8241194 TI - Peptide sequences selected by BA4, a tropoelastin-specific monoclonal antibody, are ligands for the 67-kilodalton bovine elastin receptor. AB - A 67-kDa cell-surface elastin/laminin receptor is expressed by fetal bovine ligamentum nuchae fibroblasts and neutrophils. Two hexapeptides, VGVAPG and PGAIPG, contained within hydrophobic domains of tropoelastin are binding sites for this receptor. Studies of recombinant tropoelastin proteins and synthetic peptides demonstrated that a monoclonal antibody, BA4, recognized peptide sequences similar to those recognized by the 67-kDa receptor. Taking advantage of this similarity, an "epitope library" containing random hexapeptides was screened with BA4. Four BA4-selected peptides (VGAMPG, VGMAPG, VGSLPG, and VGLSPG) were synthesized; studies of fibroblast and neutrophil migration support the hypothesis that these peptides are ligands of the 67-kDa receptor present on ligamentum nuchae fibroblasts and neutrophils. Two additional, physically similar tropoelastin peptides,AGAIPG and PGAVGP, were also identified as peptide ligands, and hence potential binding sites within tropoelastin, of the elastin receptor. These data suggest that the 67-kDa elastin/laminin receptor may interact with a wide range of structurally similar peptides containing amino acid substitutions involving small nonpolar and uncharged amino acids. PMID- 8241195 TI - Methcathinone. PMID- 8241196 TI - Magic Johnson. PMID- 8241197 TI - Skeletal maturity and growth of adolescent mothers: relationship to pregnancy outcome. AB - The purpose of this study was to evaluate the relationship between postpartum maternal bone age and the incidence of obstetric and neonatal complications in adolescent pregnancies. Bone age determinations were obtained on 93 poor, black 12- through 18-year-old adolescents during the puerperium. Results showed maternal bone ages ranging from 15 to 18 years; bone age was less than 18 years in 64 (68.8%) of the 93 adolescent mothers we studied. Maternal bone age correlated significantly with maternal chronologic age (r = 0.70) and prepregnant body size (r = 0.25) but did not correlate with total maternal weight gain and growth during pregnancy, the incidence of obstetric and neonatal complications, or infant birth weight and gestational age. Our findings suggest that many young, pregnant adolescents have the potential to grow during and after pregnancy, but do not support the hypothesis that ongoing maternal growth is an obstetric risk factor during adolescence. PMID- 8241198 TI - Advanced pubertal development and eating disturbance in early adolescent girls. AB - This study assessed the hypothesis that early adolescent girls with advanced pubertal development would be at increased risk for eating disturbance compared with less-advanced peers. Sixth-grade girls differing in levels of pubertal development responded to the Eating Disorder Inventory (EDI); to questions about weight-related concerns and dieting; and to measures of body image and appearance satisfaction. Body mass index (BMI) was calculated from measured weights and heights, and used as a measure of body fatness. Also assessed were the relative contributions of physical variables (e.g., body fatness) and psychological variables (e.g., body image) to eating- and weight-related concerns. Results supported the hypothesis that more advanced pubertal development was associated with an elevated level of eating disturbance. Analyses suggested that although normal pubertal increases in fatness and weight initially might induce eating related concerns, it is the psychological response to these changes, in the form of more negative body image, that ultimately becomes more important for promoting eating disturbance. PMID- 8241199 TI - Carnitine in adolescents. AB - Carnitine, abundant in muscle and other tissues, is normally synthesized from lysine. Its major metabolic role is transport of long-chain fatty acids into muscle where beta oxidation takes place. Normative data are available for carnitine levels, but few studies define carnitine of specific age groups. This report provides data on normal male and female adolescents in relationship to age and pubertal status. The population consisted of adolescents between 10 and 18 years, seen either for routine health maintenance or minor illnesses. Concentrations of carnitine in plasma were determined by radioisotype enzyme assay. The overall mean was 45.3 +/- 8.9 mumol/L for free carnitine. There was a trend of decreasing free carnitine with increasing Tanner stage. Among the females, there was a statistically significant correlation with both age and Tanner stage. In contrast, the data for males indicated no statistically significant correlation. The data provided standards for the evaluation of carnitine status in normal adolescents. Total carnitine in plasma ranged from 43.2 to 55.9 mumol/L and was virtually all free carnitine. The significant negative correlation of concentration of free carnitine with age and pubertal stage in females was interesting, although not of clinical importance. It suggests the possibility of female hormonal influence associated with puberty. PMID- 8241200 TI - A survey of attitudes, knowledge, and behavior related to HIV testing of adolescents and young adults enrolled in alcohol and drug treatment. AB - The objectives of this study were to assess attitudes, knowledge, and behavior related to human immunodeficiency virus (HIV) antibody testing of adolescents and young adults undergoing treatment for alcohol and drug problems. Setting and subjects were 101 adolescents and young adults enrolled in a New England substance abuse rehabilitation program. All subjects had heard about acquired immunodeficiency syndrome (AIDS) and 84% desired an HIV antibody test if it were anonymous and confidential. If found to be HIV positive, 83% stated that they would be less likely to continue substance-abuse treatment, 64% would be more likely to continue drug use, and 62% would be more likely to continue alcohol use. The likelihood of continuing school would decrease for 60%. Sixty percent would be more likely to increase use of condoms. Males were more likely than females to want to leave treatment and school, return to alcohol and drug use, and increase risk-taking behavior (p < 0.02). These findings suggest that teenagers undergoing treatment for substance-abuse problems require careful and comprehensive pre- and post-test counseling and follow-up when undergoing HIV antibody testing. Deciding when to test for HIV may impact on the success of substance-abuse treatment, as well as ability to control spread of HIV infection. PMID- 8241201 TI - Alcohol expectancies, problem drinking, and adverse health consequences. AB - This study evaluated a measure of positive and negative expected effects of alcohol and their subjective evaluation in the identification of college freshmen at high risk for problem drinking and associated morbidities. It was hypothesized that greater expectations of positive outcomes and fewer negative evaluations of negative outcomes would be associated with reports of heavier drinking and more alcohol-related health problems. College freshmen (n = 328) completed a standardized measure of expectations and subjective evaluations of positive and negative outcomes associated with drinking, and a questionnaire assessing drinking patterns and common alcohol-related health problems. Fifty-two percent of students were male and the mean age was 17.9 years (SD = 0.5). Students' expectations of positive outcomes and their subjective evaluations of both positive and negative outcomes from drinking were significantly correlated with drinking and alcohol-related health problems indices (p < 0.001). Gender, expectation of positive outcomes, and evaluation of negative outcomes explained 29% of the variance in drinking and 15% of the variance in alcohol-related health problems indices. Heavier-drinking students and those reporting more health problems expected more positive effects on their sociability and sexuality (p < 0.03) and were less concerned about cognitive and behavioral impairment as a result of drinking (p < 0.001). Students with more health problems were less concerned that drinking would lead to risk-taking or aggressive behavior (p < 0.003). Positive and negative outcome expectancies and their subjective evaluations accounted for a significant portion of the variability in drinking and alcohol-related health problems.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241202 TI - A comparative study of adolescent and adult mothers who intend to breastfeed. AB - The purpose of this study was to compare reasons for choosing to breastfeed, timing of the infant feeding decision, and intended duration of breastfeeding between adolescent and adult mothers. The study population consisted of primiparous teens and adults who were matched by ethnicity, and interviewed within 48 hours postpartum at the University of California, San Diego Medical Center from August to December, 1989. Of the 48 women in each age group, there were 40 Hispanics, 5 non-Hispanic whites, 2 blacks, and 1 Southeast Asian. Results showed that the teens differed from their adult counterparts in that they cited naturalness or convenience less often as a reason for breastfeeding. The adolescent mothers were also less likely than the adults to make the decision to breastfeed before the pregnancy, than during pregnancy or after birth. There was no difference in intended duration of breastfeeding between these two groups: the majority of women planned to breastfeed for less than 26 weeks. Early introduction of infant feeding education may serve to increase the adolescent's knowledge base from which to make the decision to breastfeed. Even though the adolescents intended to breastfeed for as long as the adults, they may benefit from increased exposure to prenatal care and childbirth classes as well as from a supportive school or work environment in order to achieve success in breastfeeding. PMID- 8241203 TI - Frequent school-based clinic utilization: a comparative profile of problems and service needs. AB - The purpose of this study is to compare frequent users of school-based clinic services with students who have an average rate of utilization. SAMPLE: Of the 1413 students enrolled in a Denver school-based clinic (DSBC) during the 1989 1990 school year, frequent clinic users (n = 73) were defined as those who visited the clinic 15 times or more (range, 15-72 visits per year). Average users (n = 82) were defined as students who visited the clinic three times during the year (the mean and median number of visits per student enrolled). Average users were compared. RESULTS: The average utilizers were found to be representative of the entire student population based on age, race, gender, and grade. The frequent users had more females (71%) and a lower grade point average (GPA) (2.11) than the average users (52% female, 2.54 GPA; p < 0.02). The frequent clinic users differed in their initial primary diagnoses as well as in the types of subsequent visits, having a significantly higher percentage of mental health-related visits than did the average users (61% versus 10%; p < 0.01). The frequent clinic users also demonstrated more high-risk behaviors such as alcohol use, sexual activity, and both family and peer relationships (p < 0.001). CONCLUSION: These data suggest that frequent clinic users have significant health needs that may require a higher rate of utilization. PMID- 8241204 TI - Neurosarcoidosis presenting as secondary amenorrhea in a teenager. AB - Sarcoidosis is a systemic inflammatory disease of unknown etiology characterized by non-caseating epithelioid cell granulomata. The lungs and reticulo-endothelial system are typically involved, and virtually any organ system may be affected. Sarcoidosis involving the central nervous system is relatively uncommon, estimated to occur in approximately 5% of patients with sarcoidosis in the United States, while the incidence throughout the world may be as high as 15%. Hypothalamic dysfunction is the most common manifestation of central nervous system parenchymatous disease in neurosarcoidosis. Polyuria and polydipsia are the most frequently occurring symptoms in patients with sarcoidosis who have dysfunction of the pituitary and hypothalamus. We describe a patient with secondary amenorrhea resulting from neurosarcoidosis involving the pituitary and hypothalamus. PMID- 8241205 TI - Sexual behavior among adolescents in Uppsala, Sweden. AB - The purpose of this study was to compare the sexual behavior of Swedish teenagers in the late 1980s with that in the late 1970s, when a similar study was performed in the same city. Another aim was to discover differences between boys and girls, smokers and nonsmokers, and pupils in theoretical and practical classes. A total of 383 high-school students answered a questionnaire about their sexual behavior, education, and attitudes. Almost one-half of the teenagers had already had intercourse. The median age at sexual debut was about 17 years. Among girls who had had intercourse, the median age at debut was lower than 10 years ago. A majority stated that their sexual behavior had been affected by the acquired immunodeficiency syndrome (AIDS) debate. This was, however, not clearly seen in their sexual behavior. Intercourse seemed to take place earlier in the relationship than 10 years ago, and the wish for more sexual experience had increased. The use of alcohol at the time of first intercourse had decreased significantly. Only 2% thought that they had too much sex education at school. As many as 41% felt they could not talk about sex with their parents. Efforts should be made both to enhance the quantity of sex education at school and to improve its quality. This might increase the chances of young people avoiding unwanted pregnancies and sexually transmitted diseases. PMID- 8241206 TI - AIDS anxieties of adolescents: determinants of "state" and "trait" anxiety dimensions in a linear structural model. AB - This study presents the effects of general psychologic characteristics on acquired immunodeficiency syndrome (AIDS) anxieties and sexual behaviour of adolescents. To this end, data were collected in a complex interview and subsequently subjected to a linear structural model analysis. The questioned adolescents were divided into one representative group (n = 256) and a second group who had participated in a voluntary human immunodeficiency virus (HIV) antibody test (n = 45). AIDS anxieties have to be divided into two independent dimensions: first, a relatively stable feeling of AIDS anxiety (trait anxiety) and second, a manifest personal anxiety toward AIDS experienced in a concrete situation (state anxiety). A principal component analysis of the primary data brought forth four variables described as depression/general anxiety, extent of phobic anxieties, compulsion, and tendency to self-consciousness. The present study reveals that the AIDS trait anxiety is more pronounced among those subjects who are not well informed about AIDS, who tend to phobic anxieties, and who observe themselves in a particularly intensive manner. The AIDS state anxiety however, is stronger among subjects who are well informed about AIDS, have sexual experience, and observe themselves intensively. Among the participants who took part in the HIV test, there were more individuals with a higher manifest AIDS anxiety and stronger tendency to depression. The percentage of adolescents who were indeed exposed to a possible risk of getting infected was relatively low. Generally speaking, those young people who are depressed, anxious, and sexually active agreed more easily to take the test than young people with a pronounced phobia toward the risk of infection and less sexual experience. As a conclusion, we can state that those adolescents with less sexual experience tend to externalize their general sexual anxieties in the form of concrete AIDS anxieties. PMID- 8241207 TI - A working formulation for the standardization of nomenclature and for clinical staging of chronic dysfunction in lung allografts. International Society for Heart and Lung Transplantation. AB - A clinically applicable, standardized method for assessing functional results after lung transplantation is required to evaluate factors that may affect long term outcome, to permit comparison of results from different centers, and to evaluate results of clinical trials. An ad hoc working group was established under the auspices of the International Society for Heart and Lung Transplantation for the purpose of developing such a clinically applicable system. Chronic allograft dysfunction is often associated with pathologic findings of obliterative bronchiolitis, the cause of which has not been defined. Physiologically, such dysfunction is associated with obstructive airways disease. The group concluded that the forced expiratory volume in 1 second was the most reliable and consistent indicator of graft dysfunction, excluding other identifiable causes. The term bronchiolitis obliterans syndrome was adopted to describe such dysfunction, recognizing that there may or may not be pathologic evidence of bronchiolitis obliterans present. Four stages of bronchiolitis obliterans syndrome were defined, each with two subcategories to indicate whether pathologic evidence of obliterative bronchiolitis had been identified. This working formulation will require reappraisal in the future to reassess its practical application and to make such adjustments as may seem appropriate. PMID- 8241208 TI - Detection of cytomegalovirus in paraffin-embedded postmortem coronary artery specimens of heart transplant recipients by the polymerase chain reaction: implications of cytomegalovirus association with graft atherosclerosis. AB - To assess the presence of cytomegalovirus in various tissues and its relevance to the development of graft atherosclerosis, 54 postmortem paraffin-embedded tissue samples from 15 heart transplant recipients who survived more than 100 days were analyzed by the polymerase chain reaction for cytomegalovirus. Eight patients had known previous cytomegalovirus exposure; 7 patients did not. Of the eight with known previous exposure, three patients (38%) died of graft atherosclerosis versus four (57%) of the seven patients without previous exposure. Of the 54 specimens, 49 were positive for beta-globin (inclusive positive control), including 38 coronary artery, five lung, five gastrointestinal, and one kidney. Only two coronary artery specimens were cytomegalovirus positive in a single patient with known cytomegalovirus exposure who did not have evidence of graft atherosclerosis at autopsy. Other tissues tested showed positive lung and stomach specimens in the patient with cytomegalovirus-positive coronary artery specimens and positive kidney, lung, and gastrointestinal specimens in a second patient. No specimens were cytomegalovirus positive in the remaining patients, despite the presence of graft atherosclerosis or previous cytomegalovirus exposure. Our data do not support the hypothesis that graft atherosclerosis is associated with latent cytomegalovirus infection of the coronary arteries. The role of cytomegalovirus in the pathogenesis of graft atherosclerosis is unknown, but possibly it represents a modulation of the immune system by remote infection. PMID- 8241209 TI - Cytomegalovirus infection and accelerated cardiac allograft vasculopathy in human cardiac allografts. AB - Cardiac allograft vasculopathy is a major limiting factor of the long-term survival of heart transplant patients. An association of cytomegalovirus infection and cardiac allograft vasculopathy has been described. We analyzed 104 endomyocardial biopsy specimens obtained from 53 heart transplant recipients and correlated the histologic findings with 115 angiograms obtained from the same patients during 4 postoperative years. The frequency of vascular changes in endomyocardial biopsy specimens was significantly higher than in angiograms during the first 3 posttransplantation years (P < 0.001, P < 0.005, P < 0.03, respectively). Also, in patients with angiographically documented cardiac allograft vasculopathy, significantly higher scores of capillary and arteriolar endothelial cell accumulation and arteriolar intimal thickness were recorded when compared with the recipients with normal angiograms (P < 0.02, P < 0.05, P < 0.005, respectively). Altogether, 29 of 53 recipients underwent cytomegalovirus infection during the first posttransplant year. Cytomegalovirus infection was associated with arteriolar endothelial cell accumulation and with increased intimal thickness of intramyocardial vessels of 1-year endomyocardial biopsy specimens when compared with cytomegalovirus-free recipients (P < 0.02 and P < 0.005, respectively). After the second year, the cytomegalovirus-associated endothelial cell response subsided, but the thickness of intima had increased when compared with cytomegalovirus-free patients (P < 0.05). Thereafter, the cytomegalovirus-associated histologic changes reached a plateau. In coronary angiography, the cardiac allograft vasculopathy changes were detected in a slower pace. Thus only after 2 posttransplantation years, cytomegalovirus-associated acceleration of cardiac allograft vasculopathy was observed, compared with cytomegalovirus-free patients (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241210 TI - Experimental cardiac allograft vasculopathy in mice. AB - Cardiac allograft vasculopathy is the most common cause of death in heart transplant recipients after the first postoperative year. The pathogenesis of cardiac allograft vasculopathy is not clearly defined. To better study this disease, a genetically well-defined and reproducible animal model such as the mouse is needed. We performed heterotopic, intraabdominal heart transplantation between two inbred strains of mice. The B10.A strain served as donors, and the B10.BR strain served as recipients. No immunosuppressive therapy was administered. The allografts in groups I (n = 6) and II (n = 6) were harvested at 30 and 50 days after operation, respectively. All allografts had palpable contractions at the time of harvest. The cardiac allografts from both groups showed mild to moderate acute cellular rejection. In groups I and II, 55% +/- 26% and 60% +/- 18% of arteries showed intimal thickening, respectively. Pathologically, the vascular lesions were characterized with varying degrees of intimal thickening, subendothelial mononuclear cell infiltration and fibrosis, frequent disruption of the internal elastic lamina, and perivascular inflammation. These findings are characteristic of cardiac allograft vasculopathy seen clinically. Isografts (n = 6) showed no vascular lesions. The heterotopic transplantation of B10.A-strain hearts into B10.BR recipients provides a useful murine model for future studies in the pathogenesis and treatment of cardiac allograft vasculopathy. PMID- 8241211 TI - Endocardial infiltrates in the transplanted heart: clinical significance emerging from the analysis of 5026 endomyocardial biopsy specimens. AB - To further elucidate the significance of endocardial infiltrates in heart transplant patients, the presence, frequency, and type of endocardial infiltrates were evaluated in 5026 endomyocardial biopsy specimens obtained from 200 heart transplant patients 0 to 75 months after heart transplantation. The relationship of endocardial infiltrates to immunologic, clinical, and demographic variables was then explored. Endocardial infiltrates were detected in 557 endomyocardial biopsy specimens (11%) from 117 heart transplant patients (58%) at 6.3 +/- 9.4 months (mean +/- SD; range, 0 to 49 months) after heart transplantation. Heart transplant patients with endocardial infiltrates were younger (p = 0.03), had a greater incidence of idiopathic dilated cardiomyopathy before heart transplantation (p = 0.05), and included a greater percentage of females (p < 0.05). Both total and treated rejection rates were significantly higher in patients with endocardial infiltrates versus those without endocardial infiltrates (p = 0.0001). Rejection on the subsequent endomyocardial biopsies was more often present in endocardial biopsy specimens with endocardial infiltrates than in those without endocardial infiltrates, both in the presence (37% versus 24%; p < 0.001) and absence (33% versus 19%; p < 0.0001) of concomitant findings of rejection. No association was identified between endocardial infiltrates and posttransplantation lymphoproliferative disorder, cytomegalovirus infection, Epstein-Barr virus infection, or cardiac allograft vasculopathy. Multivariate regression analysis confirmed that the occurrence of endocardial infiltrates is associated with rejection when adjustment is made for patient's age, gender, heart disease before transplantation, follow-up time, and number of endomyocardial biopsies after heart transplantation (p = 0.0001). CONCLUSIONS: (1) Endocardial infiltrates may occur with or without associated endomyocardial biopsy findings of rejection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241212 TI - Predictors of prognosis in patients awaiting heart transplantation. AB - Patients enrolled in a clinical heart transplantation program were evaluated to identify the predictors of prognosis in patients with advanced heart disease and to optimize timing of heart transplantation. Three hundred eighty-eight subjects were consecutively evaluated from 1985 through 1989. One hundred eighty-four patients (47.5%) had dilated cardiomyopathy; 164 patients (42.2%) had ischemic heart disease; 34 patients (8.8%) had valvular heart disease, and six patients (1.5%) had miscellaneous disorders. In each patient, 45 different parameters were considered. During follow-up (mean, 8.4 months) 166 patients underwent heart transplantation; 99 patients died (heart failure, 66 patients; sudden death, 26 patients; thromboembolism, two patients; noncardiac causes, five patients). The actuarial survival was 83% at 3 months, 77% at 6 months, 73% at 9 months, 70% at 1 year, and 59% at 2 years. The median survival time was 28 months. Analysis by Cox proportional hazard regression model revealed seven independent and significant prognostic factors: etiology (p < 0.05), NYHA class (p < 0.05), third heart sound (p < 0.05), diastolic pulmonary artery pressure (p < 0.05), pulmonary wedge pressure (p < 0.01), mean systemic blood pressure (p < 0.05), and cardiac output (p < 0.05). Cox's analysis allows the computation of patient-specific curves for predictions of residual survival time at any moment during follow-up. Moreover it can be used to calculate a simple prognostic index, which enables stratification of the patient population into three risk classes: patients at high (n = 105), intermediate (n = 160) and low (n = 123) risk of early death. Pairwise comparisons of survival between the classes were significant at 1% level. PMID- 8241213 TI - Outcome after referral for pediatric transplantation. AB - Although heart, heart-lung, or lung transplantation is performed in more than 200 children annually, the number of patients referred for such procedures is considerably greater, and little is known about the outcome of those referrals. To determine the outcome of pediatric transplant referrals we reviewed the follow up of 31 patients evaluated at our institution for heart (24), heart-lung (three), or lung (four) transplantations between January 1991 and September 1992. Indications included hypoplastic left heart syndrome (seven patients), cardiomyopathy or myocarditis (seven patients), and postoperative congenital heart disease (10 patients) for heart transplantation; Eisenmenger's syndrome (three patients) for heart-lung transplantation; and primary pulmonary hypertension (two patients), broncho pulmonary dysplasia, and cystic fibrosis for lung transplantation. Only 14 of 31 referred patients were listed for transplantation; the remaining 17 patients either improved when medical therapy was maximized (nine patients), died within days of referral (three patients), refused (two patients), chose alternate surgery (one patient), were medically unacceptable (one patient), or are currently undecided (one patient). To date 7 of 14 patients listed have undergone successful heart transplantation; the remaining seven patients either improved and did not require transplantation (two patients), refused (one patient), died waiting (one patient), are currently awaiting a donor (one patient), or underwent Norwood procedure because of donor unavailability (two patients). Thus including patients who died before listing, only 14 of 31 referrals (45%) were deemed in need of a transplant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241214 TI - Posttransplantation hemodynamics and exercise function are not affected by body size matching of donor and recipient. AB - Because the number of heart transplantations performed is limited by the number of available donor hearts, many centers have expanded the acceptable criteria for donor hearts in an attempt to provide a sufficient number of donors for the number of patients awaiting heart transplantation. Traditionally, body-size matching has been an important criteria for matching donors with potential heart transplant recipients. Although initially thought to be detrimental, studies have shown no difference in survival of patients who receive hearts from smaller donors, but heart performance in this subset of patients who receive undersized hearts has not been extensively examined. We assessed exercise capacity and 1 year posttransplantation hemodynamics in 72 consecutive adult orthotopic heart transplant recipients, grouped according to donor-recipient weight ratio and the ratio of donor to recipient body surface area. Total exercise time and relative oxygen consumption were not significantly different among three groups of patients grouped according to donor-recipient body weight ratio as follows: low, 0.60 to 0.79; mid, 0.80 to 1.0; high, more than 1.0. No difference was noted among the three donor-recipient weight ratio groups with respect to 1-year posttransplantation hemodynamics. Similarly, 1-year posttransplantation hemodynamics were not different between patients with a body surface area ratio of less than 1.0 versus those with a body surface area ratio of 1.0 or more. Differences in pretransplantation hemodynamics or graft preservation did not affect our results. Neither donor-recipient weight ratio nor body surface area ratio correlated with any posttransplantation hemodynamic measurement.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241215 TI - HLA-DR incompatibility predicts heart transplant rejection independent of immunosuppressive prophylaxis. AB - To determine whether immunosuppressive prophylaxis reduces the effect of HLA-DR incompatibility on rejection, we compared clinical and immunologic variables of patients given horse antithymocyte globulin, OKT3, or no immunosuppressive prophylaxis. Median follow-up was 27 months. Groups were similar in race; preoperative HLA reactivity; ABO matching; number of HLA-A, -B, -C, and -DR mismatches; and rejection severity. Patients given immunosuppressive prophylaxis were younger (p = 0.04), had a greater frequency of preoperative ischemic disease (p = 0.03), and had a higher 6-month rejection rate (p = 0.02). A highly significant association was found between the number of mismatches at the HLA-DR locus and rejection severity (p = 0.005). Within the OKT3-based immunosuppressive prophylaxis group and the no immunosuppressive prophylaxis group a significant association was found between the number of HLA-DR mismatches and rejection severity (p = 0.01 and p = 0.009, respectively). A similar trend was identified in the group given horse antithymocyte globulin-based immunosuppressive prophylaxis. Logistic regression, used to identify independent predictors of rejection, showed that the number of HLA-DR mismatches and not the use or type of immunosuppressive prophylaxis is significantly associated with rejection (p = 0.0009). One-year patient survival was 83% in the group with two HLA-DR mismatches and 85% in the group with one or no HLA-DR mismatch. Thus the lower rejection rates in patients with one or no HLA-DR mismatch were not associated with a 1-year survival, which was better than that of patients with two HLA-DR mismatches. The potential benefit of HLA-DR matching on rejection and patient survival must be confirmed by larger prospective studies. PMID- 8241216 TI - Analysis of deaths after heart transplantation: the University of Ottawa Heart Institute experience. AB - This study reviews the clinical outcome of the 132 orthotopic heart transplantations performed at our institute from 1984 through 1991 and focuses on the pathology of those patients who died. The study comprised 124 adults (mean age, 45.6 +/- 0.9 years) and eight children. Twenty-six adult and one pediatric deaths occurred. Operative mortality (within 30 days) was 10.6%, with 84.8% of patients surviving to discharge. Actuarial probabilities of survival at 1 and 5 years were 84% +/- 3% and 71% +/- 6%, respectively. Of the 27 deaths, six (22.2%) occurred in the operating room (from hemorrhage, graft failure, and hyperacute rejection); 14 (51.9%) occurred in-hospital after surgery (from sepsis, rejection, cytomegalovirus disease, or myocardial infarct), and seven (25.9%) occurred after discharge (from rejection and/or recurrent coronary artery disease). Two groups of patients were at higher risk: patients in cardiogenic shock requiring pretransplantation mechanical support, with in-hospital mortality of 39.1%; and patients with previous valve replacement who were taking oral anticoagulants, with intraoperative mortality of 50.0%. Pathologic examination revealed occasional instances of unsuspected coronary artery disease in the donor hearts with more than 50% stenoses of the left anterior descending coronary arteries in three of 21 (14.3%) of cases. Complications of the transplantation or related therapeutic procedures were common among those who died. The complications ranged from functionally insignificant anatomic curiosities to life threatening problems. These complications are tabulated and shown. PMID- 8241217 TI - Heart transplantation for complicated and recurrent early prosthetic valve endocarditis. PMID- 8241218 TI - Heart transplantation in patients with muscular dystrophy associated with end stage cardiomyopathy. AB - Until now, patients with muscular dystrophy and concomitant cardiomyopathy have been accepted for heart transplantation only reluctantly, because of a higher perioperative risk caused by physical disability and a suspected rapid onset of cardiomyopathy of the transplanted heart. Of a total of 582 heart transplant recipients, six patients had muscular dystrophy associated with end-stage cardiomyopathy. In three patients, preoperative neurologic diagnostic investigation revealed type Duchenne muscular dystrophy (three male); one female patient had Emery-Dreifuss; one male patient had Becker-Kiener dystrophy, and the specific classification could not be established in one male patient. Mean age was 25 years, ranging from 9 to 45 years. The postoperative follow-up ranges from 10 months to 7 years, with a mean of 40 months. All patients received triple-drug immunosuppression, consisting of azathioprine, cyclosporine, and steroids. All patients had an uneventful postoperative course. Postoperative time of intubation was not prolonged in these patients compared to other patients. All patients are physically rehabilitated. One patient died suddenly 27 months after operation. Annual recatheterization studies showed normal left ventricular ejection fraction (59.5 +/- 13.4% SD). Signs of coronary artery disease could not be observed. No progression of preexisting muscular dystrophy could be diagnosed, until now. PMID- 8241219 TI - Results of heart transplantation in recipients with active infection. AB - Although active infection in the recipient is generally considered to be an absolute contraindication for heart transplantation, seven of our first 104 transplant patients (6.7%) had active severe infection within the week before transplantation. Because of unstable clinical condition of these recipients, heart transplantation was performed when an adequate donor was offered, in spite of infection. Patients were managed with cyclosporine as the only immunosuppressive therapy during the first week after transplantation. Mortality rate and incidence of acute rejection were not different for these seven patients than for the remaining 97 patients. PMID- 8241220 TI - Orthotopic heart transplantation and concurrent coronary bypass. AB - Lack of donor organ availability is an increasing problem in heart transplantation. Methods to safely increase the donor pool are desperately needed. We report four cases of coronary artery bypass during orthotopic heart transplantation for donor hearts with normal ventricular function and subclinical coronary artery disease. An aggressive approach toward utilizing hearts from older donors with normal ventricular function may expand the donor pool and decrease the waiting period, with its attendant death, for patients awaiting heart transplantation. PMID- 8241221 TI - Expanding the donor pool: how far is too far? PMID- 8241222 TI - Directional coronary atherectomy in heart transplant recipients. AB - Cardiac allograft vasculopathy is one of the most common obstacles to the long term survival of heart transplant recipients. Percutaneous transluminal angioplasty has been used as a palliative treatment for discrete lesions caused by this disease, but it is often complicated by restenosis. This report describes two cases in which directional coronary atherectomy was used to treat the discrete lesions and explores the possible advantages of this intervention in lieu of the traditional approach. Our results suggest that directional coronary atherectomy is a feasible alternative to conventional balloon angioplasty. PMID- 8241223 TI - Serial echocardiographic evaluation of cardiac graft rejection after infant heart transplantation. AB - The effects of cardiac graft rejection on infant myocardial function as assessed by echocardiography are largely unknown. To quantitate the myocardial response to rejection, serial echocardiographic studies were prospectively performed on 20 infants (less than 1 year of age at transplantation). Two-dimensional guided-M mode tracings were digitized and quantified with a computer-assisted format. Rejection was diagnosed by clinical criteria, and 85% were graded as mild, that is without cardiac signs or symptoms. Echocardiographic analysis was blinded to rejection status, with studies available 4.2 +/- 2.9 days before rejection, on the day of rejection diagnosis, and 2.9 +/- 1.5 days after rejection treatment. Left ventricular mass increased acutely from 109% of predicted normal to 129% with rejection and decreased to 110% with therapy (p < 0.01). Left ventricular volume also tended to fall with rejection and increase with therapy. The left ventricular volume/mass ratio fell from 0.29 +/- 0.10 to 0.25 +/- 0.13 and increased to 0.37 +/- 0.15 (p < 0.05) with treatment. Systolic function was depressed by rejection as reflected in the posterior wall thickening fraction and velocity of wall thickening. Diastolic dysfunction was reflected in a decreased velocity of posterior wall thinning (-9.7 +/- 3.9 to -7.7 +/- 2.7 and recovery to -10.8 +/- 3.8 (1/second, p < 0.05) and depressed average velocity of cavity enlargement (41.2 +/- 9.6 to 36.4 +/- 8.9 and recovery to 40.7 +/- 8.6 mm/sec, p < 0.05). The utility of these echocardiographic measurements to predict rejection has not been prospectively compared with the endomyocardial biopsy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241224 TI - Registration of thoracic electrical bioimpedance for early diagnosis of rejection after heart transplantation. AB - In this study, registration of thoracic electrical bioimpedance was used for early detection of acute rejection after heart transplantation. Thirty-five heart transplant recipients were monitored by registration of thoracic electrical bioimpedance signals during the immediate postoperative period and during the outpatient follow-up. At the same time, endomyocardial biopsy specimens were taken. In addition to cardiac stroke volume index and ejection fraction, the acceleration index (seconds -2) was calculated. This acceleration index describes the acceleration of blood volume and represents a function parameter of the myocardial inotropy. Seventeen acute rejection episodes were diagnosed during the follow-up period. The average acceleration index value during the 17 rejection episodes was 92.5 +/- 11.7 sec-2 and thus significantly lower when compared with the nonrejection levels (p < 0.05). The acceleration index values decreased during acute rejections by an average of 36.4 +/- 19.3 sec-2 (28%). The sensitivity of this diagnostic parameter in the examined patients was 71%, and the specificity was 100%. Thus registration of thoracic electrical bioimpedance and calculation of the acceleration index represents a quick and noninvasive monitoring technique and can ideally be used in the outpatient clinic as a supplement to invasive endomyocardial biopsies. PMID- 8241225 TI - Neutrophil adhesion inhibition prolongs survival of cardiac allografts with hyperacute rejection. AB - Hyperacute rejection results in rapid destruction of a cardiac allograft and is characterized by infiltration of neutrophils into the donor organ. We sought to ameliorate this response by using a potent inhibitor of neutrophil adhesion to vascular endothelium, NPC 15669 (N-[9H-(2,7-dimethylfluorenyl-9-methoxy) carbonyl]-L-leucine) and determine its effect on long-term graft survival and histology. This compound specifically prevents recruitment of neutrophils at inflammatory foci by inhibiting upregulation of the CD11b/CD18 adhesion molecule located on the neutrophil surface. Lewis rats were presensitized by three serial ACI rat skin grafts placed 10 days apart. ACI rat hearts were heterotopically transplanted into Lewis recipients 10 to 14 days after the final skin graft. Group I (n = 15) was treated with a 10 mg/kg intravenous bolus of NPC 15669 before reperfusion, followed by 3 mg/kg over 30 minutes. Group II (n = 13) was given saline vehicle intravenously. Two allografts in each group were harvested at 5, 15, 30, and 60 minutes, and the remainder were followed to cessation of graft function. Early 5- to 60-minute allograft histologic findings revealed focal interstitial hemorrhage, edema, and contraction-band necrosis associated with neutrophil infiltration in group II. Group I had significant reduction of all parameters at the earliest time points. Graft survival was significantly increased in group I, 89.2 +/- 20.3 hours (mean +/- SD) compared to 27 +/- 17 hours in group II (p < 0.0001). End-stage rejection was characterized by extensive neutrophil infiltrate and hemorrhage in group II; group I grafts had a predominantly mononuclear cell infiltrate with rare neutrophils.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241226 TI - Irreversibility of cyclosporine-induced renal function impairment in heart transplant recipients. AB - The use of cyclosporine therapy for heart transplant recipients has been associated with a significant improvement of graft survival. Renal function impairment is a frequent finding in patients chronically treated with cyclosporine. The purpose of this prospective randomized study was to establish renal function in a group of heart transplant recipients receiving chronic cyclosporine treatment and to test the hypothesis of reversibility of cyclosporine-induced nephropathy by late reduction of cyclosporine. A total of 28 patients who underwent operation at least 18 months before this study began were randomly assigned to either group A (n = 14), in which the whole-blood polyclonal cyclosporine target trough level was reduced from 400 to 600 micrograms/L to 200 to 400 micrograms/L, and group B (n = 14), in which the level was maintained at 400 to 600 micrograms/L. Renal and cardiac function were assessed by paraaminohippuric acid, inulin and lithium clearances and heart catheterization, respectively, at entry and 4 months later. Cellular rejection in the transplanted heart was monitored by at least four endomyocardial biopsies every 14 days with the histologic Texas scale (grading: 0 to 10). In heart recipients renal blood flow (592 +/- 202 ml/min/1.73 m2) and glomerular-filtration rate (74 +/- 33 ml/min/1.73 m2) were significantly lower (p < 0.01), and mean arterial blood pressure (109 +/- 13 mm Hg) and renal vascular resistance (22.4 +/- 9 mm Hg/dl/min/1.73 m2) were significantly higher than the corresponding values in normal controls (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241227 TI - Role of thromboxane and angiotensin in cyclosporine-induced renal vasoconstriction in the dog. AB - Cyclosporine is associated with renal insufficiency characterized by a reduction in glomerular filtration rate that may result from renal vasoconstriction. Injection of cyclosporine in the isolated renal artery perfused at a constant flow induces a potent dose-dependent vasoconstriction of renal arterial vessels in the dog. The present study was designed to investigate the role of thromboxane A2, angiotensin, and endothelial-dependent vasodilation in the cyclosporine induced renal vasoconstriction. A specific thromboxane A2-receptor antagonist (pinane-thromboxane A2), administered at a dose of 150 micrograms, significantly decreased the renal vasoconstriction response to cyclosporine from 103 +/- 26 mm Hg to 45 +/- 11 mm Hg (p < 0.05), with cyclosporine serum levels at the end of injection averaging 382 +/- 105 and 421 +/- 150 nmol/L before and after injection of the antagonist. In contrast, pharmacologic blockade of angiotensin receptors by saralasin had no effect on the cyclosporine arterial vasoconstriction in the kidney. The endothelium-dependent vasodilation to acetylcholine was not modified during cyclosporine injection. Thus cyclosporine renal vasoconstriction appears independent of the renin-angiotensin system and of endothelium-dependent vasodilation. It is at least partly mediated by thromboxane A2. Prevention of cyclosporine vasoconstriction by thromboxane A2-receptor antagonist may likely be possible, with more potent agents having more affinity to thromboxane A2 renal receptors. PMID- 8241228 TI - Management of chronic left ventricular assist device percutaneous lead insertion sites. AB - Patients with ventricular assist devices must necessarily have percutaneous leads linking the internal device to the external console. In the chronic circumstance, the percutaneous lead insertion site may become the location of irritation or infection. At the University of Pittsburgh, a procedure has been developed for care of this site. Since the institution of the procedure, 30 patients have been mechanically supported, representing a total of 1688 patient days of support. Positive cultures were obtained from the lead insertion sites of four patients in this series, whose length of support ranged from 72 to 144 days. All four patients were hemodynamically unstable requiring support with the intraaortic balloon pump before institution of the ventricular assist device, and two patients had significant risk factors for infection development at the lead insertion site before implementation. Two of the infections were considered minor, and posttransplantation sequelae developed in only one patient. With current therapeutic protocols and the defined procedure for care of the lead insertion site, the occurrence of positive cultures associated with clinical signs and symptoms of site infection appears to be infrequent. PMID- 8241229 TI - A randomized study comparing deflazacort and prednisone in heart transplant patients. AB - Prednisone is widely used by most heart transplantation units despite its frequent side effects. Deflazacort, an oral synthetic steroid with fewer side effects, has been successfully used in patients after heart transplantation, but a prospective study comparing deflazacort and prednisone in transplant patients is lacking. We have carried out, in the last year, a prospective trial of deflazacort versus prednisone involving 35 consecutive heart transplant patients. Two of these patients died perioperatively (surgical mortality, 5.7%), and another two were excluded from the protocol because of diabetes mellitus in one patient and active infection before transplantation in the other patient. Thus 31 patients were enrolled in the 3-month study. All of them were treated with antithymocyte globulin, 10 mg/kg/day for 3 days after transplantation, azathioprine, and cyclosporine; patients were randomly assigned groups: 15 patients to receive deflazacort therapy, 1.5 mg/kg/day, and 16 patients to receive prednisone therapy, 1 mg/kg/day, starting the first day after transplantation. Steroids were rapidly tapered, reaching the maintenance dose at 2 to 3 weeks after transplantation (prednisone, 0.15 mg/kg/day; deflazacort, 0.25 mg/kg/day). Both groups were similar in terms of age, gender, ABO identity, serum cyclosporine levels, azathioprine dosage, and pretransplantation serum glucose and lipids levels. Seven endomyocardial biopsies were performed on each patient, at 1, 2, 3, 5, 7, 10, and 13 weeks after transplantation. Incidence of acute rejection was similar between prednisone and deflazacort groups; 33% of patients receiving prednisone therapy and 42% of patients receiving deflazacort therapy had one episode of 3A or higher rejection (not significant).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241230 TI - Heart xenotransplantation: lessons learned and future prospects. PMID- 8241231 TI - Heart xenotransplantation in primates. PMID- 8241232 TI - Lung transplantation in adult respiratory distress syndrome. PMID- 8241233 TI - Successful heart transplantation in a patient with histopathologically proven giant cell myocarditis. PMID- 8241234 TI - Noninvasive follow-up of episodes of significant acute heart rejection by radioisotopic methods. PMID- 8241235 TI - Successful treatment of mediastinitis and empyema in a heart transplant patient: one-stage procedure. PMID- 8241236 TI - Prostaglandin E1 in the pretransplantation period in patients with pulmonary hypertension. PMID- 8241237 TI - Harvesting of domino hearts. PMID- 8241238 TI - Undifferentiated vascular endothelial cells present in transplantation atherosclerosis display smooth muscle actin-positive cytoplasmic myofilaments. PMID- 8241239 TI - Thermodilution determination of right ventricular ejection fraction. PMID- 8241240 TI - Effects of triiodothyronine and vasopressin on cardiac function and myocardial blood flow after brain death. PMID- 8241241 TI - Interaction of sphingosine and stearylamine with phosphatidylserine as studied by DSC and NMR. AB - The interaction of sphingosine (SP) and stearylamine (SA) with dipalmitoylphosphatidylserine (DPPS) has been studied by using differential scanning calorimetry (DSC) and phosphorus nuclear magnetic resonance (31P-NMR). DSC showed that SP and SA rigidified the membranes, forming an azeotropic mixture with DPPS. The azeotropic mixture which was formed between DPPS and SP was found at a DPPS/SP molar ratio of 2:1 whereas SA and DPPS formed an azeotropic mixture at a DPPS/SA molar ratio of 1:1. An eutectic point was observed at 85 mol% of SP and 90 mol% of SA in DPPS. 31P-NMR showed the presence of a lamellar phase at DPPS/SP and DPPS/SA molar ratios lower than 1:1, whereas at higher molar ratios and at high temperatures, besides the lamellar phase, an isotropic component was detected. It was found that, at physiological pH, both SP and SA were protonated in a large extent, i.e., positively charged, since their apparent pK in the membrane were 9.1 and 8.9, respectively. The results reported in this work may be relevant to understand a number of biological effects produced by these positively charged molecules, due to their electrostatic interaction with negatively charged phospholipids. PMID- 8241242 TI - Effects of vasopressin on single Cl- channels in the apical membrane of distal nephron cells (A6). AB - We have investigated how two types of Cl- channels found in sodium transporting epithelium are regulated by arginine vasopressin (AVP). A6 cells cultured on permeable supports for 10 to 14 days have two types of Cl- channels in the apical membrane that have single channel conductances of 3 and 8 pS. In cells without AVP pretreatment, the 3 pS Cl- channel was more frequently observed than the 8 pS Cl- channel. AVP increased the open probability (Po) and single channel conductance of the 3 pS Cl- channel without significantly changing the Po or conductance of the 8 pS Cl- channels. On the other hand, AVP did not affect the number of the 3 pS Cl- channel, but increased the number of 8 pS Cl- channels. These observations suggest that AVP has two different pathways to increase apical membrane chloride conductance in distal nephron A6 cells; i.e., (1) increases the Po and single channel conductance of 3 pS Cl- channels and (2) increases the number of 8 pS Cl- channels. PMID- 8241243 TI - Chloride transport by human placental microvillous membrane vesicles. AB - Unidirectional uptake of chloride by microvillous membrane vesicles prepared from human term placentas was studied over a range of membrane potentials in the presence and absence of chloride transport inhibitors alone and in combination at maximally effective concentrations. At 0 mV, inhibition of chloride uptake by 0.1 mM DIDS, 0.5 mM DPC, and 0.5 mM DPC plus 0.1 mM DIDS was similar, suggesting a common action upon an anion exchanger; neither 0.1 mM furosemide nor 0.1 mM bumetanide alone had any effect. An inside-positive membrane potential was created by imposing an inwardly directed potassium ion gradient in the presence of valinomycin. Total chloride uptake increased with increasing membrane potential (0, 4.6, 17.3, 25.8 and 32.0 mV). The inhibition of uptake by DPC and DPC/DIDS increased with the membrane potential. The effect of DPC compared to DPC/DIDS was significantly different at 4.6, 17.3 and 25.8 mV, suggesting a degree of additivity of inhibition. Neither furosemide nor bumetanide had any effect at any potential. There was a significant increase in inhibition due to DIDS alone until the membrane potential reached 25.8 mV. But there was no significant difference between the level of inhibition at 32 mV as compared to 0 mV, providing evidence of a DIDS-sensitive conductance similar to that previously seen in patch clamp studies. We suggest that uptake of chloride across the microvillous membrane of the human placenta may be by at least three different pathways; an electroneutral, DIDS-sensitive anion exchanger, a DPC-sensitive chloride conductance and a DIDS-sensitive chloride conductance. PMID- 8241244 TI - Stabilization of egg phosphatidylcholine liposomes by the insertion of sulfatide. AB - The characteristic effects of sulfatide and its derivatives on the stability of small unilamellar vesicles of egg phosphatidylcholine liposomes in phosphate buffered saline were investigated by measuring the leakage of carboxyfluorescein that had been entrapped in these vesicles. We found that both the sulfate group and a long acyl chain, such as lignoceric acid, of the sulfatide are essential for the stabilization. The sulfatide derivatives that contain a somewhat shorter acyl chain such as stearic acid had no effect to suppress the leakage of carboxyfluorescein. The galactose residue of sulfatide is not essential to suppress the leakage. 1H-NMR study using a paramagnetic shift reagent demonstrated that the distribution of phosphatidylcholine in the vesicles containing sulfatide is homogeneous, which seems to contribute to the stability of the membrane. PMID- 8241245 TI - A 46 kDa NTPase common to rat liver nuclear envelope, mitochondria, plasma membrane, and endoplasmic reticulum. AB - A 46 kDa ATP binding polypeptide of the nuclear envelope, virtually identical to the nuclear envelope NTPase putatively involved in mRNA efflux [6], is present in all rat liver cell membranes. Its presence in nuclear envelope is not the result of cross contamination during isolation. PMID- 8241246 TI - A fluorescence anisotropy study of stabilizing effect of tri- and tetra- nitrovasodilatory drugs on DPPC liposomal membrane. AB - Glyceryl trinitrate (GT) and pentaerythritol tetranitrate (PT) are two vasodilatory drugs. The physical properties of the membrane lipid matrix, which determine the structure and function of the membrane-bound proteins, generally control the perturbation mechanism of these drugs. Thus, physical interaction of these drugs with membrane lipids is very crucial for their clinical use, different cellular processes, as well as for targetted drug delivery systems. In the present paper, we have reported for the first time the interaction between these drugs and the lipid molecules in the liposomal system of dipalmitoylphosphatidyl-choline (DPPC), as measured by steady-state fluorescence anisotropy using 1,6-diphenyl-1,3,5-hexatriene (DPH) as fluorescent probe. Our results show that by dissolving in the lipid matrix these two drugs effectively stabilise the liposomal membrane: the effect being more in case of GT than in PT, indicating that the rigidifying effect is independent of the number of nitrate groups of the two drugs. This effect increases with the increase in drug concentration, implying solubilisation of all drug molecules. Though our in vitro study has more physical significance than a physiological one, the results obtained here may be used to interpret the effects that are observed in vivo. PMID- 8241247 TI - Hepatoselective carrier-mediated sodium-independent uptake of pravastatin and pravastatin-lactone. AB - Pravastatin and pravastatin-lactone are not taken up into extrahepatic cells such as fibroblasts, or hepatoma cells such as AS-30D ascites hepatoma cells or FAO cells. In contrast, pravastatin is taken up into isolated rat hepatocytes by a carrier mediated, saturable, temperature-dependent and energy-dependent mechanism. The kinetic parameters for the saturable uptake are Km 27 microM, Vmax 537 pmol/mg per min. The permeability coefficients were determined to be 9.829 x 10(-7) cm/s at 4 degrees C, 1.76 x 10(-6) cm/s at 7 degrees C, 3.85 x 10(-6) cm/s at 17 degrees C and 5.82 x 10(-6) cm/s at 37 degrees C. The activation energy is 60 kJ/mol for 100 microM pravastatin at 37 degrees C. The Q10 values are between 1.7 and 2.8. In the presence of metabolic inhibitors and in the absence of oxygen, transport is inhibited. Uptake of pravastatin is not dependent on an extracellular to intracellular sodium-gradient. Replacement of chloride by sulfate, nitrate, gluconate or thiocyanate significantly inhibits the uptake of pravastatin. Uptake is competitively inhibited by cholate and taurocholate in the presence and absence of sodium. Pravastatin, however, competitively inhibits the uptake of cholate and taurocholate only in the absence of sodium. In addition, pravastatin-lactone enters liver cells via an energy-dependent, carrier-mediated uptake system. For the saturable energy-dependent part of the hepatocellular uptake a Km value of 9 microM and a Vmax value of 621 pmol/mg per min was determined. The permeability coefficient of pravastatin-lactone uptake is calculated to be 5.41 x 10(-6) cm/s at 37 degrees C. The uptake of pravastatin lactone is competitively-noncompetitively inhibited by pravastatin and by lovastatin and vice versa. These results indicate that the hepatoselectivity of pravastatin is due to its carrier-mediated uptake into rat hepatocytes via a sodium-independent bile acid carrier. Pravastatin-lactone resembles pravastatin sodium in its hepatoselectivity. PMID- 8241248 TI - Kinetic characterization of Na+/D-mannose cotransport in dog kidney: comparison with Na+/D-glucose cotransport. AB - Brush-border membrane vesicles (BBMV) prepared from whole dog kidney cortex, or separately from outer cortex (OC) and outer medulla (OM), were used to study the kinetics and inhibition specificity of Na(+)-dependent D-mannose cotransport. In BBMV from whole cortex the measured parameters for Na+/D-mannose uptake were Km = 0.07 +/- 0.01 mM and Vmax = 4.19 +/- 0.24 nmol/mg protein per min (n = 36). In OC BBMV the Km for Na+/D-mannose was 0.04 mM, Vmax = 3.41 nmol/mg per min. In OM the Km was 0.06 +/- 0.02 mM Vmax = 0.18 nmol/mg per min. Thus only about 5% of Na+/D mannose activity occurs in OM. Both mannoheptulose (Ki = 5.6 mM) and methyl alpha D-mannoside (Ki = 0.05 mM) are competitive inhibitors of Na+/D-mannose uptake, but at comparable concentrations have little effect on Na+/D-glucose uptake. Phlorizin is a noncompetitive inhibitor of Na+/D-mannose uptake (Ki = 4.45 microM) but a more potent and competitive inhibitor (Ki = 0.58 microM) of Na+/D glucose uptake. Phloretin (Ki = 104 microM) is a noncompetitive inhibitor of Na+/D-mannose uptake in BBMV. We conclude that Na+/D-mannose uptake is mediated by a unique high-affinity carrier located in the OC presumably at the luminal surface of the proximal convoluted tubule, with strong specificity requirements for sugars with mannose-like structures (i.e., axial C-2 hydroxyl group). Phlorizin is an inhibitor of both Na+/D-mannose and Na+/D-glucose cotransporters but is approx. 10 times less potent for the Na+/D-mannose system and also has a different mode of inhibition (i.e., noncompetitive vs. competitive). The different phlorizin inhibitory mechanisms on the Na+/D-glucose and Na+/D-mannose cotransporters may be mediated by distinct hydrophobic and sugar binding sites that characterize phlorizin-carrier interaction. PMID- 8241249 TI - Identification of two unique polypeptides from dog kidney outer cortex and outer medulla that exhibit different Na+/D-glucose cotransport functional properties. AB - The cloned Na+/D-glucose cotransporter SGLT1 and an additional recently isolated human kidney cDNA Hu14/K15 belong to a family of similar cotransport proteins including the Na(+)-dependent nucleoside and Na(+)-dependent myo-inositol carrier SMIT1. For the present study we used two different polyclonal antibodies raised against the amino acid sequence 402-420 (Ab-E) and 565-574 (Ab-P) of SGLT1 to probe brush-border membrane fractions from different regions (outer cortex- >outer medulla) of dog kidney. In Western blots both Ab-E and Ab-P react specifically (peptide blockable) with two distinct bands migrating on SDS-PAGE under reducing conditions at 75.5 kDa and 72.5 kDa. The higher molecular mass polypeptide is greatly enriched (13:1) in outer cortex and diminishes progressively towards outer medulla, whereas the lower molecular mass band is barely detectable in outer cortex but is enriched in outer medulla (4:1). Brush border membrane vesicles (BBMV) prepared from the same outer cortical and outer medullary regions that were probed with Ab-E and Ab-P exhibit strikingly different Na+/D-glucose functional transport behavior. The Na+/D-glucose cotransport activity in outer cortical BBMV is a low-affinity system with Km = 5.98 +/- 1.01 mM, Vmax = 13.05 +/- 0.55 nmol/mg protein per min, and with 1:1 Na+:D-glucose stoichiometry. Outer medulla BBMV exhibit high-affinity Km = 0.27 +/- 0.03 mM Vmax = 0.97 +/- 0.04 nmol/mg protein per min and 2:1 Na+:D-glucose stoichiometry. Comparison of SGLT1, Hu14/K15, SNST1 and SMIT indicates that Ab-E could cross react with all four, but Ab-P would recognize SGLT1, Hu14/K15, SNST1 but not SMIT. Also SNST1 is not expressed in outer cortex. Based on currently available sequence data, and its marked enrichment in outer cortex, the 75.5 kDa band is a likely candidate protein responsible for low-affinity and 1:1 Na+:D glucose stoichiometric Na+/D-glucose cotransport activity (Hu14/K15) while the minor 72.5 kDa band in outer cortex is probably SGLT1. In outer medulla, the predominant band recognized by both Ab-E and Ab-P is the 72.5 kDa protein and this could be either SGLT1 or SNST1. PMID- 8241250 TI - Ionic factors regulating the interaction of Gardnerella vaginalis hemolysin with red blood cells. AB - We have studied the hemolytic properties of an exotoxin released by Gardnerella vaginalis (Gvh). We found that hemolysis induced by Gvh is modulated by the composition of the isotonic buffer in which the red cells are suspended. In particular, low pH enhances its lytic activity, whereas low ionic strength and divalent cations diminish it. The inhibitory effects of reduced salt concentration and divalent cations occur despite normal binding of the toxin to the cells. This suggests that some post-binding step is impaired. The toxin is also able to damage cholesterol-containing lipid vesicles, and even on these model membranes it is more active at low pH. From this point of view, Gvh has some similarity to Clostridium perfringens theta-toxin, a membrane-damaging toxin belonging to the family of 'thiol-activated' cytolysins produced by Gram-positive bacteria. PMID- 8241251 TI - Quantitative analysis of proton movements associated with the uptake of weak carboxylic acids. The yeast Candida utilis as a model. AB - A quantitative analysis of the proton movements associated with the initial uptake rate of weak short-chain carboxylic acids was developed in order to estimate proton/carboxylate symports stoichiometries. The yeast Candida utilis was used as a biological model and the deduced equations were applied on the elucidation of the proton/carboxylate symports stoichiometries of lactate, succinate and citrate in a strain of that yeast species at different pH values. At pH 5.0, the proton/lactate and the proton/succinate symport stoichiometry was 1:1. In the cases of the proton/lactate and proton/citrate symports it appears that the stoichiometry ratio increased with increasing extracellular pH. PMID- 8241252 TI - Inhibition of the Ca pump of intact red blood cells by t-butyl hydroperoxide: importance of glutathione peroxidase. AB - Incubation of human red blood cells (RBCs) with t-butyl hydroperoxide (tBHP) resulted in inhibition of the Ca-pump ATPase. This was demonstrated using an assay of the Ca-pump ATPase activity in intact RBCs. In this assay, activity of the Ca-pump ATPase is expressed as the rate constant of the initial loss of ATP in RBCs exposed to Ca and A23187. Pseudo-first-order rate constants (Ca-pump ATPase rate constants) were lower in the presence of tBHP versus controls. Incubation of RBCs with tBHP resulted in both a time- and concentration-dependent inhibition of the Ca-pump ATPase (IC50 approximately 1 mM). Incubation of RBCs with tBHP also resulted in decreased oxyhemoglobin, increased methemoglobin and increased thiobarbituric acid reactive substances (TBARS). GSH levels were significantly lower in the presence of tBHP. GSH fell from a control value of 2.2 mmol/l RBC to 0.46 mmol/l RBC after incubation with 0.25 mM tBHP for 15 min. Both butylated hydroxytoluene and stobadine prevented the formation of TBARS and were partially effective in protecting the Ca-pump ATPase from tBHP-induced inhibition. Dithiothreitol was completely effective in preventing the tBHP induced formation of TBARS as well as inhibition of the Ca-pump ATPase. However, when added after exposure to tBHP, dithiothreitol was unable to restore Ca-pump ATPase activity completely. An activity of dithiothreitol independent of enzymic thiol group reduction was apparent. In the presence of mercaptosuccinate, a potent inhibitor of glutathione peroxidase, the ability of dithiothreitol to protect the Ca-pump ATPase from tBHP-induced inhibition was abolished. Therefore, protection by dithiothreitol may be afforded by its ability to replenish GSH from oxidized glutathione, thus allowing glutathione peroxidase to metabolize tBHP. These results may be interpreted to suggest that inhibition of the Ca-pump ATPase in intact RBCs occurs as a result of tBHP-induced oxidant stress and subsequent lipid peroxidation which can be prevented by certain antioxidants including butylated hydroxytoluene, stobadine, and thiol-containing compounds such as dithiothreitol. These findings provide further insight into the mode of action of hydroperoxides and certain reactive oxygen species that have been implicated in oxidative stress associated with various pathological conditions. The importance of the GSH/glutathione peroxidase system in metabolizing organic hydroperoxides is also demonstrated. PMID- 8241253 TI - Frequency domain electrical conductivity measurements of the passive electrical properties of human lymphocytes. AB - An extensive set of electrical conductivity measurements of human lymphocyte suspensions has been carried out in the frequency range from 1 kHz to 100 MHz, where the surface polarization due to the Maxwell-Wagner effect occurs. The data have been analyzed according to well-established heterogeneous system theories and the passive electrical parameters of both the cytoplasmic and nuclear membranes have been obtained. Moreover, a further analysis to take into account the roughness of the membrane surface on the basis of a fractal model yields new estimates for the membrane conductivity and the membrane permittivity, independently of the surface architecture of the cell. These findings are confirmed by measurements carried out at higher frequencies, in the range from 1 MHz to 1 GHz, on lymphocytes dispersed in both hyperosmotic and hypoosmotic media, that influence the surface complexity of the membrane due to the microvillous protrusions. The surface roughness of the cell is described by a macroscopic parameter (the fractal dimension) whose variations are associated to the progressive swelling of the cell, as the osmolality of the solution is changed. PMID- 8241254 TI - Membrane disorganization induced by perfringolysin O (theta-toxin) of Clostridium perfringens--effect of toxin binding and self-assembly on liposomes. AB - theta-Toxin (perfringolysin O) of Clostridium perfringens binds to membrane cholesterol with high (Kd approximately 10(-9) M) and low (Kd approximately 10( 7) M) affinities and causes membrane lysis of intact cells and liposomes. In order to understand the lytic process at the molecular level, the lysis of liposomes was investigated in comparison with that of intact cells. The toxin dose required to cause 50% lysis (RD50) of phosphatidylcholine/phosphatidylglycerol (82:18, mol/mol) liposomes containing 36 40 mol% cholesterol was 300-1400-times higher than the RD50 value for sheep or human erythrocytes when samples with the same cholesterol concentration were compared. However, the average number of toxin molecules bound per liposome vesicle at 50% lysis was estimated as 10-18 from the RD50 values, close to the number on erythrocytes at 50% lysis, suggesting that the number of toxin molecules adsorbed per vesicle is important for lysis. As to the toxin dose required for membrane lysis, no significant difference was observed between liposomes containing both high- and low-affinity toxin-binding sites and those containing only low-affinity sites, suggesting that theta-toxin molecules bound to low-affinity sites can assemble and cause membrane lysis as well as those bound to high-affinity sites. theta-Toxin assembles on liposomal membranes, as on erythrocytes, in a high-molecular-weight polymeric form as judged from sedimentation patterns in sucrose density-gradient centrifugation. The high molecular-weight polymers were detected only under conditions where cell or liposome lysis occurred. At low toxin doses, slower sedimenting toxin oligomers and monomers were predominant on liposomal membranes. These results indicate that toxin assembly on membranes is essential for liposome lysis as it is for cell lysis and that assembly occurs on membranes without membrane proteins. PMID- 8241255 TI - The influence of size and composition on the cholesterol mobilizing properties of liposomes in vivo. AB - As part of a study into the antiatherogenic properties of phospholipid liposomes we have investigated the capacity of a variety of preparations to increase plasma cholesterol concentrations in mice. Large unilamellar vesicles, composed of egg phosphatidylcholine, were found to be approximately twice as effective at mobilizing cholesterol than sonicated vesicles of the same composition. For egg phosphatidylcholine liposomes the change in plasma cholesterol profile is proportional to the residence time of vesicles in the circulation. Large unilamellar vesicles with a diameter of approx. 100 nm accumulate the most sterol in the animal model tested here, reaching equimolar concentrations with phospholipid after 24 h. Gel-state vesicles gave rise to a smaller increase in plasma cholesterol compared to liquid-crystalline vesicles. Our data indicate that, in vivo, net transfer of cholesterol into liposomes occurs more extensively from the lipoprotein cholesterol pool than from the erythrocyte cell membrane pool. This is consistent with the hypothesis (Williams, K.J., Werth, V.P. and Wolff, J.A. (1984) Perspect. Biol. Med. 27, 417-431) that liposomes enhance reverse cholesterol transport by generating cholesterol-poor HDL particles that can extravasate and promote more sterol efflux from peripheral tissues. PMID- 8241256 TI - Phenylarsine oxide and hydrogen peroxide stimulate glucose transport via different pathways in isolated cardiac myocytes. AB - The aim of this study was to investigate the stimulating effects of sulfhydryl reagents on glucose transport in isolated rat heart muscle cells and to compare them with the action of insulin. Low concentrations of the sulfhydryl oxidants hydrogen peroxide (H2O2) and diamide (5-100 microM), but also of phenylarsine oxide (PAO) (0.5-3 microM), that is known to specifically react with vicinal SH groups, stimulated the rate of 2-deoxy-D-glucose uptake by a factor of 4 to 8 in these cells, while higher concentrations were inhibitory. The stimulating effects of H2O2 or diamide, and, to a significantly lesser extent, those of PAO or insulin, were depressed in cells pretreated with the sulfhydryl-alkylating agent N-ethylmaleimide (56-100 microM). H2O2 raised the Vmax and lowered the Km of 3-O methyl-D-glucose uptake, while PAO or insulin solely increased Vmax. The increase in glucose transport caused by H2O2 was antagonized by the beta-adrenergic agonist isoprenaline (1 microM) or by a membrane-permeant cyclic AMP analog, whereas the effects of PAO or insulin were not altered. The action of H2O2 was additive with the stimulation induced by the protein phosphatase inhibitors okadaic acid (1 microM) or vanadate (6 mM), whereas the responses to PAO or insulin were reduced in the presence of these agents. Finally, H2O2 and PAO, but not insulin, acted additively with the protein kinase C ligand phorbol myristate acetate (0.8 microM) and with phospholipase C (0.03 units/ml). We conclude that, in cardiac myocytes, H2O2, on the one hand, and PAO (and possibly insulin), on the other hand, stimulate glucose transport via at least two distinct, SH dependent pathways. These pathways, in turn, differ from a protein kinase C- and from a phospholipase C-mediated mechanism. PMID- 8241257 TI - Macromolecular crowding effects on macromolecular interactions: some implications for genome structure and function. PMID- 8241258 TI - Transcriptional activation of Mn-superoxide dismutase gene (sodA) of Escherichia coli by MnCl2. AB - Transcription of the manganese-superoxide dismutase gene (sodA) in Escherichia coli was shown to be activated by manganese. Addition of MnCl2 increased the expression of beta-galactosidase from a sodA::lacZ protein fusion and increased the concentration of mRNA transcribed from sodA+ and sodA::lacZ constructs. The stimulatory affect of manganese on the expression of sodA::lacZ was greatly reduced (i.e., > 90%) in a strain harboring a fur mutation. We also found that manganese was capable of altering DNA topology. These results show that Mn2+ causes activation of sodA transcription. PMID- 8241259 TI - The distribution of DNA topoisomerase II isoforms in differentiated adult mouse tissues. AB - Antibodies raised against the alpha and beta isoforms of human DNA topoisomerase II were found to react with the corresponding mouse enzymes. By using these antibodies in Western blotting experiments, it was observed that these isoforms are differentially expressed in adult mouse tissues. The alpha isoform is abundant in testis and spleen. The beta isoform is also abundant in spleen and in addition is present in kidney and liver. Type II topoisomerase is low in brain and heart. These results suggest different functions for these two isoforms of DNA topoisomerase II. PMID- 8241260 TI - Identification of proximal and distal regulatory elements of the rat connexin32 gene. AB - We have investigated the genetic basis of the transcriptional regulation of the rat connexin32 gene which encodes the major gap junction protein in rat liver. Primer extension analysis of RNA isolated from adult rat liver identified multiple initiation sites clustered between -110 bp and -50 bp upstream from the translation start codon. An approx. 760 bp genomic DNA fragment upstream of the first exon which included the mRNA start sites was cloned 5' to the luciferase reporter cassette in p19LUC to yield pCx32-800/-33-LUC. Transfection of pCx32 800/-33-LUC resulted in a 200-fold increase in luciferase activity above p19LUC in the human hepatoma cell line HuH-7. Using a series of vectors containing 5' deletions of the 760 bp fragment, a basal promoter was localized between -179 bp and -134 bp. Three DNA:protein complexes were identified with the basal promoter fragment by DNA mobility shift assay using nuclear extracts from HuH-7 cells. Two of the DNA-binding complexes appeared to be related to the transcription factor Sp1. In addition, three DNase hypersensitive (HS) sites were identified within the genomic locus of connexin32 in adult rat liver. Two of the DNase HS regions behaved as silencer elements with both the native promoter and a heterologous promoter in HuH-7 cells. These data demonstrate that (1) the active promoter responsible for rat connexin32 mRNA transcription is located upstream of the first exon, (2) a basal promoter region was localized to a 50 bp region which formed multiple DNA:protein complexes, and (3) multiple proximal and distant regulatory elements are involved in the expression of connexin32. PMID- 8241261 TI - Stabilization of a protein-tyrosine phosphatase mRNA upon mitogenic stimulation of T-lymphocytes. AB - The expression of a non-receptor type protein-tyrosine phosphatase (the T-cell phosphatase or PTP-S) which shows homology with basic domains of Fos and Jun, was investigated upon mitogenic stimulation of rat splenic T lymphocytes. As studied by Northern blot analysis of total cellular RNA, mitogenic stimulation of T lymphocytes by concanavalin A resulted in an increase in the level of PTP-S mRNA; there was little or no change in the level of mRNA coding for PTP-1 (which is also a non-receptor type tyrosine phosphatase). Maximum increase of about 3-fold in the level of PTP-S mRNA occurred after 72 h of mitogenic stimulation. Mitogenic stimulation did not increase the level of PTP-S transcripts in the nucleus. The half-life of PTP-S mRNA in unstimulated lymphocytes was about 25 min which increased to 5 h after mitogenic stimulation. An inhibitor of protein synthesis, cycloheximide, increased the level of PTP-S transcripts by 6-fold in control lymphocytes but did not increase the level of PTP-1 transcripts. Treatment with cycloheximide increased the half-life of PTP-S transcripts in resting lymphocytes. The PTP-S gene product was identified as a 42 kDa polypeptide by immunoblotting. The level of PTP-S gene product increased upon mitogenic stimulation of lymphocytes by Con A and reached a maximum after 72 h, as determined by immunoblotting. These results suggest that post-transcriptional regulation of mRNA stability is an important factor in controlling the level of this phosphatase mRNA during mitogenic stimulation of T-lymphocytes. PMID- 8241262 TI - ATP-independent DNA topoisomerase from Fervidobacterium islandicum. AB - Thermotogales are thermophilic eubacteria belonging to a very slowly evolving branch in the eubacterial tree. In this report, we describe the purification and characterization of an ATP-independent DNA topoisomerase from the Thermotogale, Fervidobacterium islandicum. The enzyme, a monomer of about 75 kDa, is a type I DNA topoisomerase sharing many properties with the other bacterial topoisomerases I: it absolutely requires Mg2+ for activity, relaxes negatively but not positively supercoiled DNA and is inhibited by single-stranded M13 DNA and spermidine. A feature of the F. islandicum ATP-independent DNA topoisomerase I is its thermophily. The optimal temperature for the enzymatic activity is 75 degrees C. Studies about thermostability show that the enzyme is more stable when incubated undiluted in the storage buffer. In these conditions, 60% activity was retained after a 30 min preincubation at 75 degrees C. PMID- 8241263 TI - Analysis of the mRNA structure of the Pseudomonas putida TOL meta fission pathway operon around the transcription initiation point, the xylTE and the xylFJ regions. AB - The 13 genes encoded by the meta-cleavage operon (approx. 11 kb) of Pseudomonas putida TOL plasmid pWW0 are transcribed from a single promoter, Pm. In P. putida, transcription from Pm was strictly dependent on the presence of effector activated XylS protein. Three regions of the transcript were analyzed in the wild type strain of P. putida (pWW0) by S1 nuclease protection and primer extension analyses. A major point of transcription initiation was found in the most 5'-end of the operon, which defined a -10/-35 promoter. Another main mRNA 5'-end point located within the coding sequence of the first gene of the operon was detected and identified as a stable endonucleolytic cleavage product, which rendered this gene inactive for further translation. The estimated half-life of this leader region was 2 min. Analyses of the transcripts at xylLTE and xylFJ regions revealed the presence of several relatively stable mRNA products with 5'-ends located along these regions, which arose from specific endonucleolytic mRNA cleavage. The degradation of the stable intermediate products observed after the addition of rifampicin was similar for all the products, presenting a delay of approx. 10 min, followed by an exponential decay, with a half-life that ranged between 6 and 10 min. PMID- 8241264 TI - Molecular cloning of guinea pig CYP1A1: complete primary structure and fast mobility of expressed protein on electrophoresis. AB - Guinea pig CYP1A1 cDNA was isolated from a liver cDNA library of guinea pig treated with 3-methylcholanthrene. The cDNA, named GPc1, was 2674 bp long and contained an entire coding region for 516 amino acids. The amino acid sequence of guinea pig CYP1A1 shared 74-78% identity with those of the other mammalian CYP1A1s. RNA blot and immunoblot analyses revealed that CYP1A1 was constitutively expressed and was induced by 3-methylcholanthrene in guinea pig liver. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, guinea pig CYP1A1 expressed in yeast had a significantly smaller apparent molecular mass than expressed mouse CYP1A1. An alignment of the amino acid sequences of mammalian CYP1A1s demonstrated that guinea pig CYP1A1 was several residues shorter than the counterparts in the N-terminal region. Thus, to clarify the contribution of the N terminal sequence of guinea pig CYP1A1 to the fast mobility on the electrophoresis, mouse-guinea pig chimeric CYP1A1 was prepared through cDNA directed expression in yeast. The chimeric CYP1A1 protein had an intermediate molecular mass between mouse and guinea pig CYP1A1s indicating that the anomalous mobility of guinea pig CYP1A1 is in part due to the shortened N-terminal amino acid sequence of the protein. PMID- 8241265 TI - Primary structure and lens-specific expression of genes for an intermediate filament protein and a beta-tubulin in cephalopods. AB - Intermediate filament (IF) protein and tubulin cDNAs of cephalopod eye lenses were cloned and sequenced. The rod regions of the deduced IF proteins of the squid and octopus were more similar (68% identical) than were head (33% identical) and tail (40% identical) regions. The rod sequences were closer to squid neuronal IF protein (39% identical) than to any other known IF protein. There was only 31% identity between the rod regions, 21-30% identity between the head regions and 23-32% identity between the tail regions of the present IF proteins of cephalopods and other invertebrates. The rod regions of the cephalopod IF proteins contained the 6 heptads characteristic of nuclear lamins, consistent with an evolutionary relationship between IF proteins and lamins. The present octopus alpha-tubulin was 93% and beta-tubulin was 87% identical to the corresponding tubulins of insects and vertebrates. SDS-PAGE and peptide sequencing indicated that the order of abundance of the cephalopod lens cytoskeletal proteins was IF proteins, actin and tubulins. Northern blot hybridization revealed a 4 kb mRNA for the octopus IF protein and 2.9 and 7.3 kb mRNAs for the squid IF protein; the alpha-tubulin mRNA was about 1.8 kb in the octopus and squid, and the beta-tubulin mRNA was about 2.8 kb in the octopus. The alpha-tubulin mRNA was present in all tissues examined; by contrast, the present beta-tubulin and IF protein mRNAs appeared specialized for lens expression. PMID- 8241266 TI - Cloning and sequence analysis of the gene and cDNA encoding mouse spermidine/spermine N1-acetyltransferase--a gene uniquely regulated by polyamines and their analogs. AB - The polyamine catabolizing enzyme, spermidine/spermine N1-acetyltransferase (SSAT), has been implicated as a critical determinant of polyamine pool maintenance. SSAT has recently been shown to be positively regulated in human cell lines by polyamines and their analogs at the level of mRNA accumulation. Mouse LA-4 lung adenoma cells treated with either spermine or the spermine analog, N1,N12-bis(ethyl)spermine, produced a 2.3 and 6.5-fold increase, respectively, in SSAT mRNA. Prior evidence for transcriptional control of the enzyme prompted investigation of SSAT gene structure and its regulatory elements. The mouse SSAT gene was isolated as a 3650 bp EcoRI fragment from a lambda-J1 Mus saxicola genomic library by hybridization with human SSAT cDNA. An additional 431 bp downstream from the 3' EcoRI site were sequenced from a BamHI fragment (total gene sequence, 4066 bp). The gene contains six exons and five introns. Sequence analysis of the 774 bp of the 5' non-coding region revealed the absence of TATAA or CCAAT sequence motifs and the presence of a number of binding motifs in the 5' region of the gene with consensus binding sequences for transcription factors SP1, AP1, E2F, AP2, PEA-3 and others. The deduced amino acid sequence of the coding region differs from that of the human SSAT cDNA by five amino acids. The 527 bp of the 3' non-coding region contains four possible polyadenylation signal sites of which only one displays a typical consensus sequence. A 940 bp SSAT cDNA was isolated from Mus domesticus (BALB-C) liver lambda gt11 cDNA library. It contains a 5' untranslated region 89 bp in length and a 3' untranslated region 376 bp in length. The amino acid sequence deduced from Mus domesticus differs from that of Mus saxicola by one amino acid, from the hamster cDNA, by four amino acids and from the human cDNA by six amino acids. Further elucidation of the structural features of the SSAT gene may reveal how it is positively regulated by polyamines and their analogs. PMID- 8241267 TI - Evidence for a salt-induced conformational transition in UV-irradiated superhelical PM2 DNA. AB - Upon treatment with UV irradiation, native (supercoiled) PM2 DNA undergoes an increase in electrophoretic mobility relative to the nicked circular form in the presence of 1 M NaCl or 5 mM CaCl2 or MgCl2. This effect is dependent upon supercoiling in that the relative electrophoretic mobility decreases with decreasing superhelical density of the molecule. These findings indicate that supercoil-dependent aspects of the secondary and tertiary structure of nonirradiated PM2 DNA can be altered by a combination of UV irradiation and any of the ionic environments above. We show that the alteration is not the result of a conversion of Z-DNA segments to a right-handed helix or to a renaturation of denatured regions in PM2 DNA. Circular dichroism studies do not support a simple model in which A-form DNA induced by superhelical stress is converted to B-form DNA by UV-induced photodamage and salt. We, therefore, present three alternative explanations for these observations two of which invoke conformational transitions in secondary structure and a third which requires a change in tertiary structure due to an increase in flexibility. PMID- 8241268 TI - Transcriptional regulation of the N-myc gene: identification of positive regulatory element and its double- and single-stranded DNA binding proteins. AB - The N-myc gene is amplified and overexpressed in neuroblastoma, retinoblastoma and small cell lung carcinoma, and is considered to be related to cell proliferation and/or differentiation. The transcriptional regulatory sequences of the c-myc gene have been already identified, while those of N-myc have remained obscure for a long time. In this report, we have identified several positive and negative transcriptional regulatory elements in the upstream region of the mouse N-myc gene. Among them, an activating sequence spanning -860 to -797 bp (63 bp) could be reduced to a functional core of 21 bp from -846 to -826. This sequence, termed N21 box, worked as a positive transcriptional element when linked directly upstream (but not downstream) of the putative N-myc promoter in HeLa, not in IMR32 cells. At least two proteins, of 42 kDa and 100 kDa, bound to the double stranded N21 box, and were expressed in HeLa as well as in IMR32 cells. Moreover, the plus strand of N21 box could be specifically bound by a species of 42 kDa from either cell type and by a 37 kDa protein found only in HeLa cells. These proteins may be factors binding to positive transcriptional regulatory elements and may have a role in the regulation of N-myc expression. PMID- 8241269 TI - Heat shock and osmotically dependent steps by DNA uptake after Escherichia coli electroporation. AB - Currently, the major problem in the genetic transformation is to understand how such a large molecule as the plasmid DNA passes through the cell membrane. To solve this problem we used a simplified experimental model with Escherichia coli and the plasmid pBR322: the DNA-bacteria mixture was electroporated in a sucrose solution at 0 degree C and at fixed electrical parameters; the samples were then directly plated into agar. It was found that the electrically-induced bacterial transformation after pulsing is dependent on two factors: heat shock (delta T) and osmotic stress. Our results indicate that two mechanisms contribute to these effects: (i) thermodiffusion of the DNA across the membrane and (ii) osmotic flows of the medium, containing the DNA, into the interior of the cell. PMID- 8241270 TI - Structure of the human progesterone receptor gene. AB - The complete organization of the human progesterone receptor (hPR) gene has been determined. It spans over 90 kbp and contains eight exons. The first exon encodes the N-terminal part of the receptor. The DNA binding domain is encoded by two exons, each exon corresponding to one zinc finger. The steroid binding domain is encoded by five exons. The nucleotide sequence of 1144 bp of the 5' flanking region has been determined. PMID- 8241271 TI - Cloning and sequence of a chicken alpha-N-acetylgalactosaminidase gene. AB - A cDNA clone encoding the mature polypeptide for the chicken liver alpha-N acetylgalactosaminidase enzyme was isolated by screening a cDNA library with a synthetic sequence derived from the human alpha-N-acetylgalactosaminidase gene. The deduced amino acid sequence of the chicken liver clone was identical to the amino-terminal and cyanogen bromide derived peptide sequences obtained from the purified enzyme. The chicken enzyme has 75% identity with the human alpha-N acetylgalactosaminidase and 56-33% identity with alpha-galactosidases from several sources. PMID- 8241272 TI - Molecular cloning and characterization of cDNA encoding mouse hepatocyte growth factor. AB - A cDNA encoding mouse hepatocyte growth factor (HGF) has been cloned and completely sequenced by use of reverse transcriptase-polymerase chain reaction (RT-PCR) and subsequent cloning. Sequence analysis reveals that mouse HGF, similar to its human and rat counterparts, consists of 728 amino acids, and both the alpha- and beta-chains are encoded in a single open reading frame. Strong homology exists in the primary structure of HGF among the three species of mouse, rat and human (more than 90%), especially in Kringle 1 of the alpha chain which is assumed to be an essential domain for binding of HGF to its receptor, c-MET, a proto-oncogene product. Our results suggest the existence of evolutionary pressure to conserve the distinct structure, and presumably the biological functions, of HGF. PMID- 8241273 TI - Cloning and sequence analysis of a full length cDNA encoding human mitochondrial 3-oxoacyl-CoA thiolase. AB - The cDNA sequence of human mitochondrial 3-oxoacyl-CoA thiolase was determined. The nucleotide sequence contains an open reading frame of 1191 base pairs and encodes an amino acid sequence of 397 residues which exhibits 86.6% homology with that of the rat enzyme. Northern blot analysis gave a single mRNA species of 1.6 kb in the human liver, fibroblasts and intercostal muscle. PMID- 8241274 TI - Nucleotide sequence of the Escherichia coli K-12 transketolase (tkt) gene. AB - The gene tkt for the enzyme transketolase (TK; EC 2.2.1.1) of Escherichia coli K 12 was subcloned and the complete DNA sequence was determined from both strands. An open reading frame with 1992 bp was detected that could encode a protein with a subunit mass of 72,143. The gene forms a monocistronic operon and is transcribed counterclockwise to the E. coli chromosome. From recombinant strains, the enzyme was purified and 39 N-terminal amino acid residues were determined. The DNA-derived protein sequence showed 49.5% identical amino acid residues with the transketolase of Rhodobacter sphaeroides, 46.8% identity with the enzyme from Saccharomyces cerevisiae, and 28.9% identity with the human transketolase. PMID- 8241275 TI - Structure and expression of the mouse testicular H1 histone gene (H1t). AB - A mouse genomic library was screened with a human testicular H1 (H1t) gene fragment. One phage containing the testis specific mouse H1t histone gene and its flanking regions was isolated. Northern blot analysis showed that the mouse H1t gene is expressed only in mouse testis at the stage of pachytene spermatocytes and that the H1t mRNA is not polyadenylated. This mouse H1t gene encodes a protein which differs from the somatic mouse H1 proteins, but is similar to the known H1t proteins from rat, and man. PMID- 8241276 TI - A B cell specific immediate early human gene is located on chromosome band 1q31 and encodes an alpha helical basic phosphoprotein. AB - We have determined the cDNA sequence of a human B cell specific, immediate early gene, designated 1R20, which is inducible in response to several B cell activation signals. The cDNA sequence predicts a 196 amino acid open reading frame comprising numerous highly basic residues and the predicted structure contains several potential alpha helical domains together with eight consensus protein phosphorylation sites. The 1R20 gene has been localised by fluorescence in situ hybridisation to chromosome band 1q31, a region known to be implicated in the pathogenesis of haemopoietic malignancies. PMID- 8241277 TI - Sequence of a complete murine cDNA reflecting an S phase-prevalent transcript encoding a protein with two types of nucleic acid binding motifs. AB - Differential screening of a murine RNA-based lambda gt10 cDNA library with cell cycle phase-specific probes released a cDNA clone (lambda GS1) to a mRNA (1.8 kb) which is prevalent in the S phase of the cell cycle. The nucleotide sequence of the cDNA predicts a protein (RNPS1, 41 x 10(3) M(r) with two nucleic acid-binding domains separated by a proline-rich spacer. The N-terminal nucleic acid binding domain, about 80 amino acid residues in length, meets the requirements of an RNA recognition motif (RRM) including a perfect 'RNP-1 octamer'. The C-terminal nucleic acid binding domain spanning over 26 amino acid residues is prominent because it comprises three copies of the RRRS peptide. Domains of the latter type are considered to be involved in RNA and DNA-binding because comprised in many RNA and DNA-binding proteins. PMID- 8241278 TI - Cloning a cDNA from human NK/T cells which codes for an unusual leucine zipper containing protein. AB - A 1724 base pair (bp) clone, B3-1, was obtained from a human NK subtracted cDNA library and sequenced. The cDNA encoded a 324 amino acid protein with a calculated molecular mass of 36 kDa. The deduced protein did not contain any hydrophobic domains, suggesting that it was not secreted or membrane bound. Extensive database searches showed no significant overall homology to any known proteins or genes. The protein did, however, contain an unusually long (twice normal) leucine zipper and the nuclear targeting sequence found in many transcription factors and oncogenes. The cDNA also contained three repeats of the sequence 'ATTTA' in its 3' untranslated region, a motif associated with many oncogenes, transcription factors and interleukins. The mRNA for this gene is weakly expressed in resting NK/T cells. PMID- 8241279 TI - Sequence of the PAS8 gene, the product of which is essential for biogenesis of peroxisomes in Saccharomyces cerevisiae. AB - In a genetic screen for mutants disturbed in peroxisomal functions we found that the laboratory 'wild type' strain YP102 behaved like a typical peroxisome assembly mutant. Here, we report the sequence of the complementing gene (PAS8), coding for a protein of 1030 amino acids that appears to be a novel member of the AAA-protein family which also includes NSFp and PAS1p. PMID- 8241280 TI - The pancreatitis associated protein III (PAP III), a new member of the PAP gene family. AB - A third member of the rat pancreatitis associated protein (PAP) gene family is described here. Its messenger RNA was cloned from an intestinal cDNA library and sequenced. The encoded protein, designated PAP III, shows 66% and 63% identity with the rat PAP I and II, respectively. The PAP III gene is constitutively expressed in the small intestine and in the pancreas with acute pancreatitis, but not in the healthy pancreas. PMID- 8241281 TI - Nucleotide sequence of class-alpha glutathione S-transferases from chicken liver. AB - Two clones coding for class-alpha glutathione S-transferase were isolated from a chicken liver cDNA library. The full-length clone (933 bp) encodes a polypeptide comprising 221 amino acids with a molecular mass of 25,298 Da, including the initiator methionine. The partial clone (935 bp) encodes the C-terminal 193 amino acids of a different class-alpha glutathione S-transferase. The deduced primary amino acid sequence of the full-length clone has a 66% identical sequence with other class-alpha glutathione S-transferases. PMID- 8241282 TI - Nucleotide sequence of the genes encoding the L3, L4, and L23 equivalent ribosomal proteins from the archaebacterium Halobacterium halobium. AB - A lambda EMBL clone containing a gene cluster coding for the ribosomal proteins L3, L4, L23 and 5' region of L2 was identified in a genomic library for the halophilic archaebacterium Halobacterium halobium using a heterologous hybridization probe from the related organism Halobacterium marismortui. The clone also contains two conserved open reading frames found in H. marismortui, although with still unknown function. Its gene organization is very similar to that of 'S10 operon' of H. marismortui. The deduced amino acid sequence of these ribosomal proteins (HhaL3, HhaL4, HhaL23 and 5' region of HhaL2) shows high similarity (64-71%) to those of the archaebacterium H. marismortui and a lesser degree of similarity to their eukaryotic (31-42%) and eubacterial (17-33%) counterparts. PMID- 8241283 TI - DR-78, a novel Drosophila melanogaster genomic DNA fragment highly homologous to the DNA-binding domain of thyroid hormone-retinoic acid-vitamin D receptor subfamily. AB - Degenerate oligodeoxyribonucleotides were designed for both ends of the DNA binding domain of members of the nuclear receptor superfamily. PCR amplified Drosophila melanogaster DNA was purified and cloned (DR plasmids). Genomic lambda DASH clones were identified at high stringency with an amplified DR-78 plasmid DNA and isolated. The partial sequence shows a very probable open reading frame which would encode a peptide highly homologous to members of the thyroid hormone retinoic acid-vitamin D receptor subfamily. The fragment corresponds to a single copy gene and was mapped at position 78D of chromosome three by in situ hybridization. PMID- 8241284 TI - Cloning of LL5, a novel protein encoding cDNA from a rat pituitary library. AB - While screening a rat pituitary cDNA library for a peptide hormone receptor, we identified a cDNA that represents a novel gene. The 3.8 kb cDNA has an open reading frame of 2.3 kb encoding a protein of 781 amino acids (M(r) = 87,507). Southern blot analysis of rat, mouse, bovine and human genomic DNAs revealed that a homologous gene is present in these species probably in a single copy. Northern blot analysis showed that in addition to the pituitary gland, the gene is also expressed in other rat tissues. Scanning of DNA and protein databanks revealed no significant homology to any other sequence. Thus, this gene encodes a heretofore unidentified protein. PMID- 8241285 TI - Disease processes in epithelia: the role of the actin cytoskeleton and altered surface membrane polarity. AB - The establishment and maintenance of cell polarity is essential for normal epithelial function. Disruption of the underlying processes, either as a primary inborn defect or as a secondary result of other pathologic processes, can lead to loss of epithelial polarity and further cellular and organ-level dysfunction. Continued elucidation of the processes involved may prove fruitful both in the understanding of basic cell biology and in the understanding and treatment of a variety of disease states. PMID- 8241286 TI - Effect of the nephritogenic autoantibody of Heymann's nephritis on plasminogen binding to gp330 and activation by urokinase. AB - Previous results have shown that the autoantibody eluted from the glomeruli of rats with active Heymann nephritis contain a population of antibodies not only to the putative autoantigen of the disease, gp330, but also to plasminogen. Since gp330 has been shown to serve as a receptor for plasminogen, we have analyzed the effects of autoantibody on plasminogen-binding to gp330 and activation of plasminogen to plasmin by urokinase. Autoantibody does not inhibit the binding of plasminogen to gp330. The binding of autoantibody to plasminogen was shown to be very specific for the compact conformation of glu-plasminogen. The change in the conformation of plasminogen when its lysine-binding sites are occupied or after conversion to plasmin results in a significant decrease in autoantibody-binding. The most significant effect of autoantibody on this system is the inhibition of plasminogen activation to plasmin by urokinase. The binding of autoantibody to plasminogen acts as a competitive inhibitor of the reaction by apparently blocking access of urokinase to plasminogen's activation site. These results indicate that autoantibody obtained from the immune deposits in the glomeruli of rats with active Heymann nephritis does not inhibit the binding of plasminogen to gp330 but does significantly alter the urokinase catalyzed activation of plasminogen to plasmin. PMID- 8241287 TI - A study on in vitro glycation processes by matrix-assisted laser desorption ionization mass spectrometry. AB - The number of glucose molecules condensed on glycated bovine serum albumin have been easily determined by means of matrix-assisted laser desorption/ionization mass spectrometry. Measurements were carried out on samples from incubation of the protein with glucose at different concentrations (0.02 M, 0.2 M, 2 M and 5 M). A clear increase in molecular mass of BSA with respect to incubation time is detected. In contrast to what is observed with fluorescence, the plots of molecular mass increase vs. incubation time show the occurrence of a steady state, corresponding to the complete saturation of all the protein sites reactive against glucose. Comparison of fluorescence and molecular mass data reveals that some further reactions, different from condensation, must take place, which could be in principle either intramolecular or originated by reactivity of modified condensed glucose moieties vs. free glucose. PMID- 8241288 TI - Control of sorbitol metabolism in renal inner medulla of diabetic rats: regulation by substrate, cosubstrate and products of the aldose reductase reaction. AB - Streptozotocin diabetes induces a 4-fold increase in the maximal velocity of inner medullary aldose reductase as determined in vitro but increases sorbitol synthesis in intact inner medullary collecting duct (IMCD) cells only 1.3-fold. In order to resolve this discrepancy we investigated the importance of intracellular factors in controlling the role of cellular sorbitol synthesis. These factors include glucose concentration, sorbitol concentration, the activity of the NADPH-regenerating pentose phosphate pathway, intracellular NADP and NADPH content, and intracellular reduced (GSH) and oxidized glutathione (GSSG). It was found that the apparent Km of cellular sorbitol production for glucose was identical in control and diabetic rats (56 +/- 18 vs. 59 +/- 14 mmol/l D glucose), whereas Vmax increased by 31% in diabetes. In inner medullary collecting duct cells of diabetic rats containing 146 +/- 5 mumol sorbitol/g protein, sorbitol synthesis was slightly lower (-15%), compared to cells which had been sorbitol-depleted prior to the experiment (87 +/- 4 mumol sorbitol/g protein). However, no inhibitory effect of sorbitol (up to 200 mmol/l) was observed on aldose reductase activity in vitro. In diabetic rats the content of NADPH was about 32% lower than in the control rats (3.8 +/- 0.3 vs. 5.6 +/- 0.4 mumol/g protein) and the ratio of NADPH/NADP was decreased from 25.6 +/- 5.1 to 8.6 +/- 1.7. In homogenates of the inner medulla the activity of 6-phospho gluconate dehydrogenase (EC 1.1.1.43) was identical in both experimental groups, so the pentose phosphate shunt seems to be unaltered. GSH content in diabetic rats was also diminished (4.02 +/- 0.67 mumol/g protein vs. 7.41 +/- 0.5 mumol/g protein) and the GSH/GSSG ratio fell from 92.6 to 57.4. In enzyme tests in vitro an apparent Km of 7.3 +/- 1.9 mumol/l of the aldose reductase for NADPH was found; NADP acted as competitive inhibitor with an apparent K(i) of 183 +/- 31 mumol/l. Aldose reductase activity was also found to be strongly inhibited by the SH-group reagent p-chloromercurybenzoesulfonate (apparent K(i) = 0.85 x 10(-6) mol/l). Combining the results obtained on the properties of the aldose reductase in vitro and the observation made in the intact cells, the investigators suggest that the decrease in NADPH/NADP ratio, as well as changes in the redox state in the cells of diabetic animals, can play a significant role in the control of sorbitol synthesis. PMID- 8241289 TI - A disorder of collagen biosynthesis in patients with cerebral artery aneurysm. AB - 12 patients with subarachnoid hemorrhage due to rupture of a cerebral aneurysm were examined clinically for symptoms and signs of a connective tissue disorder and biochemically for details of the biosynthesis of collagen. No uniform clinical pattern of any connective tissue disorder was seen in these patients, although selected signs were observed. Skin fibroblast cultures were then established. The rate of procollagen production in two cell lines was reduced by 40% and 50%, respectively, and the intracellular accumulation of hydroxy[14C]proline (as a percentage of total hydroxy[14C]proline) was increased by 70% in each relative to eight control cell lines. No difference was found in the degree of intracellular degradation of procollagen. After pulse-labelling, however, the radioactive procollagen was secreted into the medium in 1 h in the control cells, but required at least 3 h in the two aneurysm patient cell lines. The results, thus, suggest that delayed secretion of procollagen rather than increased intracellular degradation led to the reduction in the rate of procollagen synthesis in these two fibroblast lines from patients with cerebral artery aneurysm. PMID- 8241290 TI - L-2-hydroxyglutarate dehydrogenase: identification of a novel enzyme activity in rat and human liver. Implications for L-2-hydroxyglutaric acidemia. AB - In this paper we studied the degradation of L-2-hydroxyglutarate in tissues from rat and man in order to try and find the underlying basis for the accumulation of this metabolite in L-2-hydroxyglutaric acidemia patients. The results show that L 2-hydroxyglutarate is not degraded by an oxidase but via a dehydrogenase which was found to be present in liver only. This newly identified enzyme activity was characterized kinetically, although the nature of the reaction product remains to be identified. PMID- 8241291 TI - A 31P and 1H-NMR investigation in vitro of normal and abnormal human liver. AB - Spectral changes in human hepatic tumours and possible systemic effects of tumour on host liver were assessed by 31P and 1H in vitro NMR spectroscopy. The 1H and 31P spectra from liver tumour biopsies showed significant elevation in phosphoethanolamine, phosphocholine, taurine, citrate, alanine, lactate and glycine, and significant reduction in GPE (glycerophosphoethanolamine), GPC (glycerophosphocholine), creatine and threonine compared to histologically normal tissue. 31P-NMR spectra obtained from histologically normal tissue within tumour bearing livers showed significant elevation in phosphoethanolamine and phosphocholine compared to data from liver biopsies from nontumour-bearing patients (pancreatitis). These results suggest that alterations in membrane metabolism in host liver can be detected by 31P-NMR. PMID- 8241292 TI - Aflatoxin B1-binding proteins in primary cultured hepatocytes of chicken embryo: studies in vivo and in vitro. AB - Among several aflatoxin B1 (AFB1)-binding proteins in primary cultured hepatocytes of chicken embryo, a cytosolic 25 kDa protein proved to be an AFB1 specific binding protein. Binding of AFB1 to a cytosolic protein of the same molecular mass as that seen in vivo was also detectable in a cell-free system and was dependent on the microsomal components. The binding was observed in the in vitro systems derived from AFB1-sensitive animals such as chicken, rabbit, and rat. These findings suggest that the 25 kDa AFB1-binding protein(s) is involved in the intracellular process of AFB1-mediated toxicity. PMID- 8241293 TI - Hb Villaverde [beta 89 (F5) Ser-->Thr]: the structural modification of an intrasubunit contact is responsible for a high oxygen affinity. AB - Hb Villaverde [beta 89 (F5) Ser-->Thr], identified in a Spanish patient, is a new human hemoglobin variant, electrophoreticaly silent, responsible for a severe erythrocytosis. This abnormal hemoglobin displays a very high oxygen affinity and a markedly reduced cooperativity that is partly restored in the presence of IHP. Determination of the structural abnormality was achieved on a mixture of the normal and abnormal beta-chains. After isolation of the abnormal tryptic peptide by RP-HPLC, its sequence was determined by mass spectrometry. The structural abnormality disturbs the intrasubunit interaction between helices F and H and, thus, may weaken the C-terminal bonds of the deoxy conformation and the heme contacts of several hydrophobic residues. Hb Villaverde demonstrates that this intrasubunit contact between helices F and H is essential for the cohesion of the hemoglobin molecule. PMID- 8241294 TI - Tissue distribution of cytochrome c oxidase isoforms in mammals. Characterization with monoclonal and polyclonal antibodies. AB - Monoclonal and polyclonal antibodies specific to the two isoforms of subunit VIa of bovine cytochrome c oxidase were generated and used to study the tissue distribution of this subunit pair in beef, human and rat. The so-called H-(heart) form was found exclusively in heart and skeletal muscle, whereas the so-called L (liver) form was the only isoform present in brain, kidney, liver and smooth muscle. Little or no L-form was detected in skeletal muscle. In bovine heart no subunit VIa-L was detected, while in human heart the subunit VIa-H and VIa-L isoforms were present in roughly equal proportions. These results imply that, in humans, the deficiency of a subunit VIa isoform may have a different effect on the physiology of heart then on the physiology of skeletal muscle. PMID- 8241295 TI - [Indications for spiral-CT in the upper abdomen and the retroperitoneum]. AB - Spiral CT allows the continuous scanning of different organ volumes. This series illustrates our experiences gained with spiral CT technique imaging of the upper abdomen and retroperitoneal space in about 150 patients. The scanning parameters were 120 kvp, 165 mA with a slice thickness of 8 mm and a table speed of 8 mm/s. The total scanning time was 32 s. If required, a non-ionic iodinated contrast medium was administered via a medial cubital vein or the upper mesenteric artery by performing an arterial portography. CT during arterial portography for preoperative detection and localization of focal liver lesions, staging of renal cell carcinoma, and imaging of pancreatic disease are at present the main indications for using the spiral CT technique. Furthermore, it is the method of choice for scanning uncooperative patients. PMID- 8241296 TI - [Depiction of evoked regional brain activity using magnetic resonance tomography]. AB - After brief summary of the various parameters determining contrast behaviour in diagnostic magnetic resonance imaging, special attention is paid to recent developments in its application to the study of brain perfusion and oxygenation. With and without extrinsic contrast media, it is now possible to image regional tissue perfusion and brain oxygenation and thus the evoked activation of regions of the brain with a high spatial resolution of about 1 mm and a temporal resolution of about 50 ms. The blood oxygenation dependent magnetic properties of hemoglobin are used as sensitive endogenous indicators of the oxygenation state of blood. In dependence upon evoked local brain activity, contrast changes of up to 30% have already been observed. Possible future developments with respect to the application of MRI to even more complete functional studies of the brain and other organs will be mentioned. PMID- 8241297 TI - [Magnetic resonance tomographic characterization of intracranial tumors by evaluating the blood-brain barrier and regional cerebral circulation]. AB - The aim of this prospective study was to characterize intracranial tumors on the basis of the degree of blood-brain-barrier (BBB) disruption and tumor blood flow (TBF). We studied 28 patients with brain tumors by MRI. BBB disruption was demonstrated by a pathological increase of signal intensity in T1-weighted spin echo images (1.5 T, TR = 600 ms, TE = 10 ms, alpha = 90 degrees) after intravenous gadolinium-DTPA injection. TBF flow was assessed by an MRI-method based on the signal intensity decrease in T2-weighted gradient-echo images (TR = 25 ms, TE = 20 ms, alpha = 10 degrees) immediately after gadolinium-DTPA bolus injection. Typical constellations include intake BBB and low TBF in low-grade intraaxial tumors (astrocytoma I/II), disrupted BBB and heterogeneous TBF in high grade intraaxial tumors (glioblastomas and metastases), and disrupted BBB and high TBF in extraaxial tumors (meningeomas). This study demonstrates the quasi simultaneous assessment of the blood-brain-barrier and tumor blood flow. The results support the concept that additional uncorrelated information is obtained from the assessment of regional cerebral blood flow that may be helpful for the differential diagnosis of brain tumors. PMID- 8241298 TI - [Thrombolysis in acute cerebral ischemia]. AB - During the last years some medical centers have performed emergency treatment for intracranial thromboembolic occlusions by thrombolysis using streptokinase, urokinase, and recombinant tissue plasminogen activator intraarterially or intravenously. So far, it appears that the risk of intracranial hemorrhage with clinical deterioration is not increased by thrombolysis if thrombolysis is performed during the first 8 hours after symptom onset. Successful recanalization of occlusions in the anterior or posterior circulation is associated with a decrease of mortality and morbidity. Intraarterial application of the thrombolytic agent seems to be more effective than intravenous application. However, there is still a lack of controlled studies that could exclusively prove which thrombolytic agent and which mode of application is the most effective with regard to arterial recanalization and clinical outcome. PMID- 8241299 TI - [The possibilities of diagnosing the act of swallowing in contrast medium aspiration]. AB - 19 patients with contrast agent aspiration were examined prospectively utilizing a combined approach of an orally administered low-osmolal and non-ionic contrast agent and a digital X-ray system with a dynamic technique (8 images/s). This combined technique enabled a sufficient assessment of the aspiration mechanisms in all patients. PMID- 8241300 TI - [Radiologic findings in renal metastases]. AB - US and CT findings in 41 cases of renal metastasis with known primary malignancy, mostly lymphoma and pulmonary carcinoma, are analysed retrospectively. All of them were examined with US and 22 with CT. The renal metastases were bilateral in 20 cases and multiple in 21. The echo structure was "infiltrative" in 20, non infiltrative in 21, homogeneous in 11 and non-homogeneous in 30. 33 metastases were echo-poor, 5 were hyperechoic, two iso-echoic and one of mixed echogenicity. The lymphomas were usually homogeneous, infiltrative and echo-poor, whereas the metastases of pulmonary primaries were usually nonhomogeneous, non-infiltrative, echo-poor and some hyperechoic. None of hyperechoic metastases were lymphomas. One case out of 22 examined with CT indicated a false negative US finding, while 4 US examinations showed only some of the lesions detected with CT. The lesions were hypodense and non-homogeneous in plain CT and gave non-homogeneous enhancement in all cases except for 4 with homogeneous lesions. CT showed smaller and more numerous lesions than US. PMID- 8241301 TI - [Jejunogastric invagination: plain radiologic, sonographic and computed tomographic findings]. AB - Jejunogastric intussusception is a rare complication after gastric surgery and when its diagnosis is delayed a high mortality rate can be expected. This paper reports on the characteristic findings of this entity at ultrasound and CT examination. Both diagnostic procedures have a high sensitivity. Sonography is the method of first choice because the diagnosis can be made with a high grade of certainty. CT allows the differentiation of the distinct stages of the disease and the views given by CT are often more easily accepted by the surgeons. PMID- 8241302 TI - [Generalized clostridial infection in a 60-year-old patient with massive soft tissue emphysema on thoracic radiography]. AB - We report a case of a 60-year-old patient with progressive soft tissue emphysema caused by infection with clostridium septicum. In contrast to a rather linear spread of air in non infectious soft tissue-emphysema, in this case a mainly vesicular spread of air in the soft tissue is noted on plain films. Together with the clinical history, this finding may indicate an infectious cause. The radiological interpretation is an important step in the diagnostical workup. PMID- 8241303 TI - [Perirenal metastasis of a malignant melanoma in a young child]. AB - A perirenal metastasis of a malignant melanoma in a young child can accompany a big tumor volume with extensive calcifications and tumor necroses. The characteristics of the tissue on computed tomography or ultrasound did not allow the certain differentiation of this situation from tumors of the adrenal or tumors of the kidney. PMID- 8241304 TI - [Magnetic resonance tomography in a case of the Tolosa-Hunt syndrome]. AB - We report a rare case of Tolosa-Hunt syndrome in 48-year-old woman with terminal renal insufficiency and acute onset of headache and disturbances of the cranial nerves. MRI was performed to exclude cerebral disease. 14 days later the patient died from the terminal renal insufficiency. Neurohistopathological examination revealed pseudotumorous tissue components confirming the diagnosis of Tolosa-Hunt syndrome. PMID- 8241305 TI - [Population of triplet-excited states of tryptophanyls and tyrosyls in proteins]. AB - The relative limiting concentrations of triplet excited states of tryptophan and tyrosine residues have been measured by the ESR method. The magnitude of limiting concentration of the tryptophan and tyrosine residues T1-states does not vary to fit their numbers in some proteins. A limitation on the concentration of T1 states is accounted for by the fact that in protein molecules there occurs the radiationless excitation energy transfer of chromophores on residues in the triplet state. PMID- 8241306 TI - [The effect of an inhibitor on the dependence of single-enzyme reaction rate on pH]. AB - The enzyme reaction speed dependence on pH was calculated. It was shown that two speed maxima and higher pH sensitivity should be in reaction with inhibitor. PMID- 8241307 TI - [Interconversion of two possible forms of endothelium-derived relaxation factor- S-nitrosocysteine and an dinitrosyl complex of iron with cysteine]. AB - Interconversion of S-nitrosocysteine (cys-NO) and dinitrosyl iron complex with cysteine (DNIC-cys) has been observed in 15 mM Hepes-buffer (pH 7.4) which was added with cys, Fe2+ and NO. Cys-NO was mainly detected in this solution at 37 degrees C. The content of DNIC-cys was predominant in frozen solution. The destruction of DNIC-cys with 0.5 HCl or desferrioxamine resulted in the formation of cys-NO. The hypothesis was advanced that endothelium-derived relaxing factor (EDRF) is the equilibrium mixture of DNIC-cys and cys-NO. EDRF which forms in vascular endothelial cells is presented in these cells mainly as DNIC-cysteine, because endogenous cys destroys cys-NO. When EDRF appears in intercellular space with low content of cys it transforms to cys-NO. PMID- 8241308 TI - [A new approach to the study of statistical properties of genetic sequences]. AB - An approach to the study of the properties of genetic texts is proposed. It is based on the investigation of the frequencies of all possible words (subsequences) in a text. The most important effect is that the original text could be reconstructed completely without deletions and/or mistakes using the set of words which are met in the text as a single copy. The length of words for which the effect occurs is a measure of the text redundancy. Some real genetic sequences were studied as well. PMID- 8241309 TI - [Physical properties of the DNA-water system]. AB - The review of original and literary data obtained by the different physical methods on the structure, mechanical, electrostatic and energetic properties of DNA-ions-water system was done. On the basis of the examined results the main problem of DNA structure formation its dynamic and stability were discussed. PMID- 8241310 TI - [Modification of the lipid membrane matrix with magnesium]. AB - The influence of sulphuric magnesium on the molecular regulation processes in lecithin-water systems was investigated by the optical methods of polarized microscopy, refractometry and infrared spectroscopy. The obtained changes of structure parameters are interpreted by the modification role of Mg2+ cations and SO4(2-) groups. PMID- 8241311 TI - [The effect of human and rat erythrocyte volume on the structure of hemoglobin protoporphyrin]. AB - Changes of erythrocyte hematoporphyrin at different cell volumes were investigated using resonance Raman spectroscopy. Correlation between changes of the position and intensity ratio of 1260 cm-1 bands RRS spectrum and cell volume was observed. Changes of this kind suggest that electronic density redistribution occurs between the C=C and C-N bonds of porphyrin molecule. The redistribution of the electronic density can cause extension of the "core" and increase of the distance between the nitrogen and iron atoms. The iron atom displaces to the macrocycle plane causing decrease of its spin number. PMID- 8241312 TI - [Regulation of human erythrocyte volume. The role of calcium channels activated by calcium]. AB - A mathematical model of stabilization of erythrocytes volume which takes into account the homeostasis of univalent ions and the calcium regulation of potassium flux through the membrane has been studied. The model has the only stable solution at all actual values of parameters. The investigation of the steady state cell volume dependence on membrane permeability for potassium and sodium ions has shown that volume is much better protected against a nonspecific change in permeability than against a specific increase in the membrane permeability for potassium or sodium ions. In the case of a nonspecific increase of the erythrocyte membrane permeability for cations the cell volume has best stabilization when the stoichiometry of sodium and potassium ions transported by a Na,K-ATPase is 3/2. PMID- 8241313 TI - [Turbidity of myofibril suspensions]. AB - Turbidity (optical density) of isolated myofibril suspensions was investigated at different concentrations, light wavelengths and solid angles of the spectrophotometer photocell. The turbidity is shown to be strongly dependent on the condition of its measurement which does not permit to obtain correct values suitable for theoretical interpretation. PMID- 8241314 TI - [Aggregation and sedimentation of erythrocytes in a magnetic field]. AB - The system of equations describing aggregation and settling of erythrocytes or other weakly magnetizable aggregating particles in an external constant inhomogeneous magnetic field is investigated. The magnetic force applied to erythrocytes was assumed to be the basic mechanism of field action on the blood. It is shown that the magnetic field with a special configuration may decrease the sedimentation time and increase the suspension stability to small perturbations. PMID- 8241315 TI - [Blastocoele potential in the early frog embryo]. AB - The resistance of intercellular gaps between the blastomeres and the potential of the blastocoel cavity in the cleaving frog eggs were estimated by means of microelectrode technique. The maximal and minimal values of these parameters were evaluated in different phases of the mitotic cycle. PMID- 8241316 TI - [Mathematical modeling of thermal regulation in local hyperthermia]. AB - A mathematical model for description of transient heating of biological tissue during the local hyperthermia is proposed. The model is based on the bioheat transfer equation and on equation of thermoregulation which represents integro differential relationship between the local temperature and the local blood perfusion rate. One-dimensional electromagnetic heating of a semi-infinite homogeneous tissue volume and a local approximation of the problem are studied numerically and analytically. A possibility of the oscillatory local temperature response in return to local constant power heating is investigated for different forms of the thermoregulation equation. It is shown that long-time undamped temperature oscillations are predicted under assumption about memory's mechanism in blood flow regulation is accepted. PMID- 8241317 TI - [Analysis of parameters of an electrostatic field, arising upon electric stimulation of the cat visual cortex]. AB - Calculations of the locality of cat's areas 17 and 18 under electrical stimulation are given. The electrical conductivity anisotropy of cortical layers is taken into account. PMID- 8241318 TI - [Surface charge of isolated hepatocyte nuclei during change in their functional activity]. AB - We have measured densities of surface charge of hepatocyte nuclei during hydrocortisone induction and activation induced with amino acids mixture. The value of single nucleus charge, calculated from the value of charge density, changes more during both forms of induction, that the value of the electrokinetic potential. Thus the former better characteristics the electrokinetic properties of nuclei during their functional swelling. PMID- 8241319 TI - [Photoreactivation of Ca2+-ATPase and Mg2+-ATPase after exposure to ionizing radiation]. AB - Plasma membranes of thymocytes in doses 10(2), 10(3) and 10(4) Gr were irradiated and subsequently illuminated by lambda = 445,5 nm or lambda = 518 nm light. Then the activity of Ca(2+)-ATPase and Mg(2+)-ATPase was determined. The photomodulation of the electron shell of calcium and magnesium atoms is a factor decreasing the effect of radiation on Ca(2+)-ATPase or Mg(2+)-ATPase. PMID- 8241320 TI - [Mathematical model of self-regulating parasitic system]. PMID- 8241321 TI - Transforming growth factor beta and fibroblast growth factor as promoters of tumor progression to malignancy. AB - Polypeptide growth factors are a diverse group of biological regulators. Because they are fundamentally involved in the cellular processes that are important for transformation and progression to malignancy, alterations in growth factor control and in their signal pathways are often observed in tumor cells. In this review, we consider the participation of growth factors and the mechanisms by which they effect tumor progression, using as examples members of the transforming growth factor beta (TGF-beta) and fibroblast growth factor (FGF) families. We explore the hypothesis that although abrogation of TGF-beta negative growth regulation is necessary for transformation, in the later stages of tumor progression, TGF-beta plays a direct role in the enhancement of invasion and metastasis as an autocrine stimulator of these processes. In addition, we present evidence that demonstrates both the potential and the importance of members of the FGF family in transformation and induction of metastasis. Several models of growth factor regulation of malignancy are presented in which we demonstrate (1) a link between TGF-beta 1 mitogenic stimulation of malignant cells and alterations in the expression of ribonucleotide reductase, a key rate-limiting step in the synthesis of DNA and in cell proliferation; (2) autocrine and/or intracrine FGF mitogenic stimulation of malignant cell proliferation and metastasis; and (3) autocrine TGF-beta regulation of malignant cell locomotion and invasion through elevated proteolytic activity and increased synthesis of hyaluronan and RHAMM, a novel hyaluronan cell surface receptor. PMID- 8241322 TI - Resistance to inhibition of cell growth by transforming growth factor-beta and its role in oncogenesis. AB - Transforming growth factor-beta (TGF beta) was first described as a soluble factor capable of transforming normal rat kidney cells in the presence of transforming growth factor-alpha (TGF alpha) or epidermal growth factor (EGF). TGF beta is now known to have a wide spectrum of effects, depending on cell type, culture conditions, and the presence of other growth factors. Whereas picomolar concentrations of TGF beta completely inhibit the growth of most nonmalignant epithelial cells and early hematopoietic progenitor cells, a review of the published English language literature reveals that virtually all malignant cells of epithelial or hematopoietic origin are refractory to the antiproliferative effects of TGF beta in vitro. When compared with normal epithelial cells, malignant carcinoma cells require significantly higher doses of TGF beta to achieve a similar degree of growth suppression or they are only partially suppressed by large doses of TGF beta. This loss of sensitivity to TGF beta is associated with the development of invasive properties. More than one genetic event appears to be required to convert a sensitive cell into one that is completely refractory to TGF beta. In conclusion, resistance to the growth inhibiting effect of TGF beta appears to be a late but consistent event in the process of malignant transformation in vitro. Identification of the genetic lesions that result in the loss of response to TGF beta may lead to the design of new therapeutic modalities to arrest tumor growth. PMID- 8241323 TI - Gap junctional communication and neoplastic transformation. AB - Gap junctional communication (GJC) is mediated by channels consisting of connexins and can be differentially regulated by ions, second messengers, kinases, phosphatases, and cell adhesion molecules. Tumor cells and oncogene transformed cells often, but not always, show reduced homologous GJC between themselves. A more stringent correlation may exist between transformation and reduced heterologous communication between transformed cells and normal neighbors. Reduced GJC seems to stimulate tumor promotion but has no significant effect on the initiation phase of carcinogenesis. These effects may reflect the importance of intercellular passage of second messengers or other small molecules in cell growth control. Some evidence suggests that gap junction in combination with cell adhesion molecules can affect metastatic potential, but a clear picture has not yet emerged. Coupling and gap junction expression can be regulated both pre- and posttranslationally in oncogene-transformed cells. Src probably downregulates GJC in fibroblasts by tyrosine phosphorylation of connexin43. The Ras-induced reduction in GJC appears to be caused by decreased connexin expression. E1A, but not Myc and Fos, downregulates GJC to some extent. Artificial expression of connexin in glioma, hepatoma, chemically transformed, and src-transformed cells can restore GJC and suppress growth and/or tumorigenesis. These results argue for involvement of GJC in transformation and growth control. PMID- 8241324 TI - How and when do tumor cells metastasize? AB - Tumor cells metastasize when they have successfully passed a number of invasion steps. We hypothesize that each step is a microecosystem, the basic elements of which are neoplastic cells, host cells, and extracellular matrix (ECM). We review here molecular pathways with a regulatory function in these microecosystems: homotypic cell-cell adhesion molecules counteracting invasion; complexes of lytic (pro-)enzymes, their receptors and inhibitors regulating focalized breakdown of the ECM; matrix components, their cellular receptors and motility factors governing cell migration; heterotypic cell-cell adhesion molecules initiating extravasation, encompassing in the vasculature the sequence: cell arrest, ECM lysis, cell migration; factors allowing survival and growth at ectopic sites. We conclude that delicate molecular balances within microecosystems are responsible for the temporary and repeated invasion events leading to metastasis. PMID- 8241325 TI - Bioelectrical impedance analysis: a promising predictive tool for nutritional assessment in continuous ambulatory peritoneal dialysis patients. PMID- 8241326 TI - Serositis: comparative analysis of histological findings and pathogenetic mechanisms in nonbacterial serosal inflammation. AB - Peritonitis is the established term for infective inflammation of the peritoneum, while serositis generally refers to nonorganismal inflammation in any serous cavity, including the peritoneum. In continuous ambulatory peritoneal dialysis (CAPD) literature, however, culture-negative peritoneal inflammation is referred to as "sterile" or "chemical" peritonitis. These terms not only imply unwarranted etiologic assumptions, but may also deflect attention from the existence of medical conditions to which the peritoneum is subject. This is evident in CAPD literature where there is little recognition that the peritoneum, as a member of the serosa and a secretor of lamellar bodies, is prey to a wide range of disorders. Thus before, during, and after CAPD, the membrane is liable to fall victim to disease states unconnected with the process of dialysis. Significant peritoneal pathology occurs as part of a pan-serositis, which may be metabolic (uremia, cholesterolosis), autoimmune (systemic lupus erythematosus, rheumatoid disease, acute rheumatism, endocrinopathies), genetic (recurrent hereditary polyserositis), allergic (eosinophilic serositis), and granulomatous in nature. This paper presents a comparative analysis of histopathological presentation and pathogenetic mechanisms involved in all forms of peritoneal serositis. It incorporates recent advances in molecular biology of the membrane into a holistic reappraisal of peritoneal pathology, revealing hitherto unrecognized homologies in peritoneal reaction to diverse disorders. PMID- 8241327 TI - Quantitation of lymphatic drainage of the peritoneal cavity in sheep: comparison of direct cannulation techniques with indirect methods to estimate lymph flow. AB - OBJECTIVE: It has been suggested that lymphatics may contribute to ultrafiltration failure in patients on continuous ambulatory peritoneal dialysis (CAPD) by absorbing dialysate and ultrafiltrate from the peritoneal cavity. In most studies lymphatic drainage has been estimated from the disappearance of an instilled tracer from the peritoneal cavity or estimated from the appearance of an intraperitoneally administered tracer in the bloodstream. However, in sheep it is possible to cannulate several of the relevant lymphatics that drain the peritoneal cavity and assess lymph drainage parameters directly. The purpose of this study was to estimate lymph drainage from the peritoneal cavity in sheep using the disappearance of tracer from the cavity and the appearance of intraperitoneally instilled tracer in the bloodstream and to compare these results with those obtained from our previous studies using cannulation techniques. DESIGN: Experiments were performed in anesthetized and nonanesthetized animals. Volumes of 50 mL/kg of Dianeal 4.25% containing 25 microCi of 125I-albumin were infused into the peritoneal cavity. RESULTS: In anesthetized sheep the calculated peritoneal lymph drainage from monitoring the disappearance of tracer from the peritoneal cavity over 6 hours was 1.873 +/- 0.364 mL/kg/hour. Monitoring the appearance of tracer in the blood gave significantly lower peritoneal lymph flow rates of 1.094 +/- 0.241 mL/kg/hour. Directly measured lymph flow rates from our earlier publication were lower still and ranged from 0.156 +/- 0.028-0.265 +/- 0.049 mL/hour/kg, depending on how we estimated the right lymph duct contribution to peritoneal drainage, since we could not cannulate this vessel. We repeated these experiments in conscious sheep. The value for lymph flow estimated from the disappearance of tracer from the peritoneal cavity was 2.398 +/- 0.617 mL/hour/kg and from the appearance of tracer in the blood, 1.424 +/- 0.113 mL/hour/kg. The lymph flow rates monitored from indwelling lymphatic catheters ranged from 1.021 +/- 0.186-1.523 +/- 0.213 mL/hour/kg (again, depending on our estimates for the right lymph duct). CONCLUSIONS: Lymph flow rates measured from indwelling lymphatic catheters provided the most conservative values for lymphatic drainage of the peritoneal cavity under dialysis conditions. Estimates of lymphatic drainage based on the appearance of tracer in the blood gave values that were on average higher. The method using the disappearance of tracer from the cavity to estimate lymph flows overestimated peritoneal lymph drainage. Fluid was lost from the peritoneal cavity, and the estimated proportion of liquid lost through lymphatic drainage depended on the technique used to measure lymph flow rates. PMID- 8241328 TI - Peritoneal transport during dialysis with amino acid-based solutions. AB - OBJECTIVE: To evaluate the potential clinical role of amino acids as an osmotic agent. DESIGN: The peritoneal transport of fluid, amino acids, and other solutes was investigated during a 6-hour single-cycle peritoneal dialysis with PDA 1% versus 1.36% glucose (n = 6) or PDA 2.7% versus 3.86% glucose solution (n = 9). PATIENTS: Fifteen stable nondiabetic continuous ambulatory peritoneal dialysis (CAPD) patients. RESULTS: The fractional absorption of the osmotic agents at 6 hours was higher with PDA 2.7% versus glucose 3.86% (p < 0.005). The diffusive mass transport coefficient, KBD, calculated for a period of dialysate isovolemia was higher with PDA 2.7% versus PDA 1% for essential, nonessential (p < 0.005), and total (p < 0.05) amino acids. The intraperitoneal volume-over-time curves and KBD values for urea, creatinine, glucose, albumin, beta 2-microglobulin, and total protein did not differ between the amino acid solutions and the corresponding glucose solutions. KBD for urea was significantly higher during the dwell with PDA 2.7% versus PDA 1% (p < 0.05). Plasma amino acid concentrations increased substantially during the first 1-2 hours and then decreased gradually. Valine and methionine rose to 792% and 1119% of baseline values, respectively. CONCLUSIONS: We conclude that the peritoneal transport of fluid and investigated solutes, except amino acids, was not different with the amino acid solutions compared with the corresponding equimolar glucose solutions. However, ultrafiltration tended to be lower with amino acid solutions. Furthermore, the fractional absorption of amino acids and KBD values for amino acids was higher with PDA 2.7% versus PDA 1%, suggesting an effect of the hypertonic amino acid solution on the peritoneal membrane transport properties. Also, the hypertonic PDA 2.7% solution yielded nonphysiologically high plasma levels of several amino acids. We therefore consider this solution not to be safe enough for long-term clinical use. PMID- 8241329 TI - Choline incorporation into phospholipids in mesothelial cells in vitro. AB - OBJECTIVE: To determine the effect of extracellular choline concentration on phospholipid production and handling by peritoneal mesothelial cells in vitro. DESIGN AND MEASUREMENTS: Radiolabeled choline was used to monitor the formation of phosphatidylcholine (PC), sphingomyelin (SPH), and lysophosphatidylcholine (LPC) by rat and rabbit mesothelial cells as a function of concentration and time of exposure to choline. The subcellular location of the newly formed phospholipids was examined by ultracentrifugation in Percoll-sucrose gradients using analytical cell fractionation techniques. The fatty acid composition of the PC formed was determined by thin-layer chromatography (TLC) and gas chromatography. RESULTS: Choline incorporation into PC, SPH, and LPC increased with extracellular choline levels up to 640 mumol/L, which is 100 times greater than physiological levels of choline in plasma and 20 times higher than choline levels measured in peritoneal dialysis effluent. The newly formed, radiolabeled phospholipids were primarily found in a single subcellular compartment that exhibited a buoyant density of 1.05 g/mL in Percoll-sucrose gradients. Analysis of the fatty acyl groups of PC obtained from the mesothelial cells showed enrichment in palmitic [16:0], oleic [18:1], and linoleic [18:2] acids. CONCLUSION: The rate of phospholipid formation by mesothelial cells in vitro can be manipulated, in part, by choline concentration. PMID- 8241330 TI - Short-term clinical study with bicarbonate-containing peritoneal dialysis solution. AB - OBJECTIVE: The evaluation of the efficacy, adequacy, clinical tolerance, and safety of a new bicarbonate continuous ambulatory peritoneal dialysis (CAPD) solution. DESIGN AND PATIENTS: A 6-week cross-over clinical study in 6 stable CAPD patients was performed. After a control period (2 weeks) with a standard CAPD solution (lactate, 35 mmol/L), a two-chamber bag containing 34 mmol/L of bicarbonate was used for 4 weeks. A breakable valve divided the two chambers, one containing bicarbonate and the other calcium. The two solutions were mixed just before use, thus avoiding the calcium and magnesium carbonate precipitation. RESULTS: No differences between control and study periods were found for blood urea nitrogen, creatinine, total proteins, albumin, total and ionized calcium, phosphate, sodium, potassium, chlorine, and hemoglobin. Blood bicarbonate significantly increased from 21.25 +/- 2.02 to 23.36 +/- 1.15 (p < 0.05) during the study. The peritoneal equilibration tests for urea nitrogen, creatinine, proteins, sodium, potassium, and glucose were slightly reduced during bicarbonate dialysate, but this effect was compensated for by a slight increase of ultrafiltration, thus keeping peritoneal clearances constant. Residual renal function did not change during the study. No side effects occurred during the bicarbonate period. CONCLUSION: A CAPD bicarbonate solution is effective in uremic acidosis correction, does not affect dialysis adequacy, is safe, and well tolerated. PMID- 8241331 TI - Enhanced ultrafiltration in rabbits with bicarbonate glycylglycine peritoneal dialysis solution. AB - OBJECTIVE: The aim of the study was to investigate net ultrafiltration (NUF) with a new bicarbonate glycylglycine (BiGG) peritoneal dialysis solution compared to the standard lactate (La) solution. DESIGN: In six groups of 12 normal rabbits each we measured NUF after a 2-, 4-, and 6-hour peritoneal dialysis with a BiGG solution (pH 7.35) and a standard La solution (pH 5.5) of similar glucose, electrolyte, and osmolality formulation. Furthermore, we studied the phosphatidylcholine concentration in the effluent of the two solutions. RESULTS: NUF volume was significantly greater with the BiGG rather than with the La solution by approximately 15% (p < 0.05), 30% (p < 0.01), and 40% (p < 0.005) at 2, 4, and 6 hours, respectively. The glucose absorption rate was greater with the La solution than with the BiGG solution, but the difference was significant only at 2 hours (p < 0.05). pH was increased in the La solution from its initial value of 5.5 to 7.18, 7.32, and 7.40 at 2, 4 and 6 hours, respectively, while it remained almost unchanged in the BiGG solution. Phosphatidylcholine (PC) in the peritoneal effluent was significantly higher in the BiGG solution in all instances (p < 0.0001). CONCLUSION: It is concluded that the BiGG solution, which has a stable pH, 7.35, due to the potent buffering capacity of bicarbonate and glycylglycine, enhances peritoneal NUF by maintaining a higher osmotic gradient and retarding lymphatic absorption through an increase in PC concentration in the peritoneal cavity. PMID- 8241332 TI - Improvements in continuous ambulatory peritoneal dialysis technique survival: further experience from a U.K. renal unit. PMID- 8241333 TI - Cimetidine improves the accuracy of clearance measurements in peritoneal dialysis patients. PMID- 8241334 TI - Cost analysis of dialysis modalities for pediatric acute renal failure. PMID- 8241335 TI - Surreptitious use of antimicrobial agents by CAPD patients. PMID- 8241336 TI - Preliminary experience with a vaccination against Staphylococcus epidermidis in patients on chronic peritoneal dialysis. PMID- 8241337 TI - Dialysate leak and hemoperitoneum after laparoscopic cholecystectomy in a CAPD patient. PMID- 8241338 TI - Peritonitis due to Wangiella dermatitidis in a patient on CAPD. PMID- 8241339 TI - Normal function from a Tenckhoff catheter after a 7-year rest. PMID- 8241340 TI - Recurrent hydrothorax following repeated pleurodesis using autologous blood. PMID- 8241341 TI - Cloudy dialysate due to adenocarcinoma cells in a CAPD patient. PMID- 8241342 TI - A case of CAPD peritonitis due to Hemophilus influenzae. PMID- 8241343 TI - Evaluation of a suprapubic mass in a CAPD patient. PMID- 8241344 TI - Literature. October-December 1993. PMID- 8241345 TI - Detection of p53 mutations using nonradioactive SSCP analysis: p53 is not frequently mutated in myelodysplastic syndromes (MDS). AB - p53 is one of the most frequently mutated genes in human cancers. Since p53 has been implicated in lymphatic and some myeloid leukemias, such as the blastic phase of chronic myelogenous leukemia, we sought to address the role of p53 gene mutations within exons 4-9 in myelodysplastic syndromes (MDS), a myeloid preleukemic condition. In order to avoid the potential hazard of using radioactive single-strand conformation analysis (SSCP), we used a nonradioactive SSCP method based on the silver stain of small minigels. In cell lines with known point mutations of the p53 gene, aberrant migrating bands were found. Serial dilutions indicated a sensitivity comparable to radioactive methods. Furthermore, a common polymorphism within the 4th exon of the p53 gene was easily detected. However, of 17 primary samples from patients with MDS, none harbored a p53 gene mutation. We conclude that this nonradioactive method can easily be used to screen for p53-gene mutations, and that p53-gene mutations do not play a major role in the pathogenesis of MDS. PMID- 8241346 TI - Idarubicin is active on MDR cells: evaluation of DNA synthesis inhibition on P388 cell lines. AB - Multidrug resistance is frequently found in patients affected by hematological malignancies and has been related to a poor prognosis of acute leukemia. In the present paper we report results concerning the activity of idarubicin, an anthracycline derivative, on the leukemic P388 and P388 doxorubicin-resistant cell lines. The results clearly show that idarubicin inhibits DNA synthesis in the resistant cell line more actively than doxorubicin. PMID- 8241347 TI - High PAI activity with correlation to triglyceride and HDL cholesterol values in patients with coronary artery disease with no difference in survivors of myocardial infarction. AB - The fibrinolytic capacity of blood depends mainly on the amount of tissue-type plasminogen activator (t-PA) activity and plasminogen activator inhibitor type-1 (PAI-1) activity. Previous studies linked high PAI activity or low t-PA activity with the development of atherosclerosis and thromboembolic diseases. Yet, there are conflicting reports in the literature as to whether there is higher PAI activity in patients with myocardial infarction (MI) than in patients with coronary artery disease (CAD) without previous MI. In this retrospective study, t PA activity, t-PA antigen, and PAI activity before and after a venous occlusion test (VOT) of 10 min were assessed in 109 patients with angiographically documented CAD, in two subgroups of CAD patients with (n = 66) or without (n = 43) previous MI, and in subgroups of CAD patients according to their triglyceride levels and other risk factors. The mean values of t-PA activity in the whole patient group showed a 100-fold increase and a 3.1-fold increase in t-PA antigen after VOT (0.03 +/- 0.03 to 3.0 +/- 6.8 U/ml and 16.5 +/- 6.9 to 51.0 +/- 25.4 ng/ml, p < 0.05). PAI activity was 24.4 +/- 11.0 before and 19.6 +/- 13.2 U/ml after VOT. Within the CAD group, no difference was found between patients without MI and survivors of previous MI in PAI activity before VOT (24.6 +/- 10.7 vs. 24.3 +/- 11.3 U/ml) and after VOT (19.0 +/- 12.1 vs 20.0 +/- 14.0 U/ml), or t-PA activity before (0.03 +/- 0.01 vs. 0.04 +/- 0.04 U/ml) and after VOT (2.8 +/- 7.0 vs. 3.2 +/- 6.7 U/ml). In 39.4% of CAD patients elevated plasma PAI activity before VOT (> 25 U/ml) was found. This subgroup of patients represented the highest PAI activity after VOT (p < 0.05), the lowest t-PA activity after VOT (p < 0.001), and the highest triglyceride levels (p < 0.05). In 11% of the patients, a small increase in t-PA activity (less than 0.5 U/ml) after VOT was seen. This group showed the lowest t-PA antigen after VOT (p < 0.001) and the highest fibrinogen level (p < 0.05). Both subgroups showed the same distribution among patients with and without MI. CAD patients with triglyceride levels over 200 mg/dl had the highest PAI activity values before VOT (28.3 +/- 11.8 U/ml; p < 0.01) and after VOT (24.9 +/- 13.2 U/ml; p < 0.01), resulting in low t-PA activity after VOT (p < 0.01).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8241348 TI - Reflex sympathetic dystrophy after bone marrow transplantation. AB - We report on three patients of 201 who, after bone marrow transplantation (BMT), developed reflex sympathetic dystrophy (RSD). The syndrome occurred on days +40, +43, and +147, respectively, without known causes of RSD. The ankles, the feet, and the knees were affected. Long-lasting immobilization, slow bone marrow recovery, and other BMT-related factors could have predisposed to RSD. PMID- 8241349 TI - Spontaneous IL-2 production in vitro in two patients with pure red cell aplasia. AB - We investigated spontaneous cytokine production in two patients with pure red cell aplasia (PRCA). The peripheral blood mononuclear cells (PBMNC) from two patients produced IL-2. Cyclosporin A (CyA) suppressed in vitro IL-2 production in one patient, but not in the other. Spontaneous IL-2 production disappeared in one patient 10 months after the start of CyA therapy. The patient for whom CyA therapy was stopped after the disappearance of spontaneous IL-2 production has remained in continuous remission for 1 year. The present case suggests that spontaneous IL-2 production in PBMNC might be an indicator of disease activity. PMID- 8241351 TI - Impacts on the future practice of craniofacial surgery. PMID- 8241350 TI - A 72-year old man with low-grade non-Hodgkin's lymphoma, cold agglutinin hemolysis, rapidly progressive jaundice, hepatomegaly, and dyspnea. PMID- 8241352 TI - Cleft palate speech--an opinion. PMID- 8241353 TI - Meningiomas of the anterior skull base expanding into the orbit, paranasal sinuses, nasopharynx, and oropharynx. AB - The natural progression of the infiltrative anterior skull base meningioma, expanding into orbit, paranasal sinuses, nasopharynx, and oropharynx during 12 years is presented. The development of the lesion is demonstrated on computed tomography. The patient was operated on six times from the age of 14. Pathological investigation revealed infiltrative meningoteliomatous meningioma without signs of malignancy. PMID- 8241354 TI - Craniofacial neurosurgical approach for extensive hyperostotic meningioma. AB - We present a patient with a craniofacial meningioma that had been considered inoperable for 8 years but has now been successfully treated using the combined neurosurgical and craniomaxillofacial approach. The tumor extensively involved the meninges, skull, face, left orbit, and maxilla. Vision and mild proptosis of the left eye had worsened over the last several years. Computed tomography and magnetic resonance imaging scans revealed an extensive intraosseous meningioma with intracranial and extracranial extension. An angiogram revealed an intensely vascularized mass supplied by a markedly enlarged ophthalmic artery. The tumor was approached through a bicoronal incision. The meningioma that extended into the dura was removed entirely from the brain. The left orbital roof and the lateral and medial walls were resected along with the meningioma, which extended through the orbit into the maxilla. The defect in the dura was closed with a pericranial flap. Split parietal bone and lyophilized cartilage were used for reconstruction of the cranial and orbital defects. Three and a half months after the initial surgery, further reconstruction was performed using calvarial, rib, and iliac bone and lyophilized cartilage. A methylmethacrylate implant was inserted but then removed due to chronic wound drainage. It was substituted with demineralized perforated bone implants from Pacific Coast Tissue Bank. Seventeen months after the initial surgery, there was no evidence of the tumor. PMID- 8241355 TI - Factors for osseous repair and delivery: Part II. PMID- 8241356 TI - Use of Medpor porous polyethylene implants in 140 patients with facial fractures. AB - Use of alloplastic materials in facial bone reconstruction is still controversial. Medpor porous polyethylene is a relatively new implant material that is well suited for this purpose and has a number of advantages over other alloplasts. It is a pure polyethylene with a unique manufacturing process and pore size. Technically, it is easy to work with; it can be carved, contoured, adapted, and fixated to obtain a precise three-dimensional construct. Physically, it is a pure, biocompatible, strong substance that does not resorb or degenerate. It demonstrates long-term stability, high tensile strength, resistance to stress and fatigue, and a virtual lack of surrounding soft-tissue reaction. Rapid tissue ingrowth occurs into the pores. Extensive vascular ingrowth creates the potential to transport cellular products that fight infection deep into the implant. The implant was used in 140 patients from June 1988 to August 1991 to treat acute orbitozygomatic injuries (71), acute Le Fort injuries (24), delayed orbitozygomatic injuries (33), and delayed onlay augmentation (12). In this series, there was 1 instance of implant infection requiring removal, and no implant migration, or exposure. PMID- 8241357 TI - Two patients with multiple facial clefts including cleft lip and palate. AB - Report of 2 patients with cleft lip and palate and multiple facial clefts are described using the anatomical classification of Tessier. The first patient was severely polymalformed, with heart, vertebral, and facial malformations. He was examined clinically and with an experimental three-dimensional computer reconstruction that identified an association of a classic cleft lip and palate with cleft nos. 4 and 11 on the right side and cleft nos. 3 and 11 on the left. The second patient, a partial form, with cleft lip and palate and cleft nos. 3 and 11 on the right side, also presented with bilateral cleft no. 7, preauricular tags, and an important microgenia. This second patient was operated for the cleft lip using the principles of Millard and Onizuka with a supplementary Z plasty. We comment on the difficulties of delimitation and treatment when the clefts are very close together and the difference in "quality" of the tissues compared with classic cleft lip and palate. PMID- 8241358 TI - Management and complications of middle- and upper-third facial compound injuries: an Iranian experience. AB - Surgery for 772 patients with middle- or upper-third facial injuries was carried out between 1984 and 1990 in two university hospitals in Tehran. The most common etiological factor was firearm injuries (69.04%; n = 533), followed by road traffic accidents (24.44%; n = 189). The most common associated nonfacial injuries were head injuries (36.97%; n = 285 followed by ocular injuries (23.18%; n = 179). Treatment of middle- or upper-third facial injuries ranged from minor repair to major surgery with autogenous or allogenous grafts. The complications associated with middle- or upper-third facial injuries were either postinjury complications or postoperative complications and ranged from minor infection to loss of vision. PMID- 8241359 TI - The study of the modern human face through radiography: Part II. An anthropological concept. PMID- 8241360 TI - Surgical techniques of cranial vault expansion for increases in intracranial pressure in older children. AB - Standard craniofacial techniques to expand the cranial vault were successful in treating elevations of intracranial pressure (ICP) in 7 older children. Of the 7 patients, a diagnosis of craniosynostosis was made in 6 and deformational head deformity with slit ventricle syndrome in 1. There were 5 boys and 2 girls, whose ages ranged from 3 1/2 to 8 years (mean, 5.2 yr). Three patients had previously undergone surgical treatment of craniosynostosis. One patient presented with visual changes and papilledema. Another with a deformational skull deformity was shunt-dependent and was diagnosed with slit ventricle syndrome. This patient had undergone several temporal craniectomies to control recurrent symptoms of increased ICP. The remaining 2 patients appeared to have craniosynostosis as an explanation for reduced cranial vault size and elevated ICP, but the specific sites of sutural fusion could not be identified. Total follow-up ranged from 3 to 18 months, and evidence of resolution of signs and symptoms of increased ICP was achieved in each of the 7 patients. Plastic surgeons involved in caring for children with craniosynostosis should be particularly aware of the possibility of increased ICP developing or recurring following craniofacial surgery. PMID- 8241361 TI - Distraction of the frontal bone outside the cranial plane: a rabbit model. AB - Distraction of the frontal bone outside the cranial plane using the Ilizarov principle was performed in 17 22-week-old New Zealand white rabbits. Five rabbits had frontal bone osteotomy only and were in the control group; 5 rabbits were placed in the sham control group and had frontal bone osteotomy plus application of a customized headgear appliance; and 7 rabbits were placed in the experimental group, which underwent frontal bone osteotomy application of the headgear and distraction. The frontal bone was elevated 1 mm every other day for a period of 8 weeks, and the animals were then killed. Cephalometry was performed both preoperatively and at the end of the 8-week period. Histological examination of the skulls was also performed. The experimental group showed a significantly elevated frontal bone compared to the sham control group (p < 0.05). Callous bone filled the distracted segment, which united the frontal bone with the cranial plane. Therefore, frontal bone advancement by distraction osteogenesis is possible using this rabbit model. PMID- 8241362 TI - Repair of nonossifying defects in the skull. PMID- 8241363 TI - Neuropeptide Y increases force development through a mechanism that involves calcium entry in resistance arteries. AB - The hypothesis that neuropeptide Y (NPY) potentiates noradrenaline (NA)-induced vascular force development by increasing free intracellular Ca2+ ([Ca2+]i) was tested in rat mesenteric resistance arteries. NPY (100 nM) alone was not able to increase either the contraction or [Ca2+]i. However, pretreatment of mesenteric resistance arteries with 100 nM NPY potentiated both [Ca2+]i and active stress induced by 1.5 microM NA. Addition of 100 nM NPY to vessels that had been precontracted with NA (1.5 microM) elicited a large increase in [Ca2+]i and an increase in active stress development. In Ca(2+)-free medium containing 2 mM ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid, the potentiating effect of NPY on the NA-induced contraction was prevented, and readdition of Ca2+ resulted in a large increase in both [Ca2+]i and active stress development. It is concluded that NPY potentiates NA-induced contraction in the isolated mesenteric resistance artery by inducing a rise in [Ca2+]i through an influx of Ca2 from the extracellular source. PMID- 8241364 TI - Phorbol ester-induced contractions of swine carotid artery are supported by slowly cycling crossbridges which are not dependent on calcium or myosin light chain phosphorylation. AB - This study determined if phorbol ester-induced contraction of vascular smooth muscle requires calcium-dependent myosin light chain (MLC) phosphorylation and, if not, whether the mechanical characteristics of the contraction in terms of stiffness and crossbridge cycling are similar to those during a calcium- and MLC phosphorylation-dependent contraction. Carotid arterial strips were exposed to 1.0 microM phorbol 12,13-dibutyrate (PDBu) in the presence of normal physiological salt solution (PSS) or after calcium depletion in calcium-free PSS and compared with contraction elicited by calcium-containing 110 mM KCl-PSS. PDBu induced maximal stress in both the presence and absence of calcium. While there was a temporal correlation between MLC phosphorylation and shortening velocity during KCl depolarization, shortening velocity was dissociated from MLC phosphorylation during PDBu stimulation. The stress-stiffness relationship was not different during KCl and PDBu stimulation, suggesting similar crossbridge interactions even though MLC phosphorylation levels were significantly different. These results demonstrate that PDBu-induced contraction of the swine carotid artery is not dependent on calcium or MLC phosphorylation. We suggest the possibility that activation of a calcium-independent PKC isoform may result in the expression of an inherent level of actin-activated myosin ATPase activity resulting in the slow development of stress. PMID- 8241365 TI - Quantifying the effect of locally delivered anticoagulant drugs: modification of an in vivo model of venous thrombosis. AB - Increasingly, attention is focusing on the local delivery of antiplatelet and fibrinolytic therapy as a means of preventing intravascular thrombosis. A simple, reproducible model of thrombosis, based upon vascular damage is needed to test these agents in vivo. We have therefore modified a rat vena cava model of venous thrombosis based upon vascular injury and stasis. Wistar rats are anaesthetised, the inferior cava dissected free and a segment isolated by slings distally above the iliacs and proximally above the left renal vein. All other tributaries are ligated. Vascular injury is induced by externally applying soft-jaw clamps for 5 min. Agents to be tested are introduced into the isolated segment via a left renal vein cannula left in situ for 15 min and then flushed from the cava. Blood is allowed to refill the segment, all remaining slings are tied and the animal left for 30 min before being killed. The cava is then opened and thrombus removed and weighed. Scanning electron microscopy of the cava after clamping shows areas of normal endothelium interspersed with areas of denuded endothelium and exposed subendothelial connective tissue. Histological and immunohistochemical staining indicates the thrombus is composed of red cells, platelets and fibrin. The model was validated by assessing the effects of 2 different doses of locally delivered tissue-type plasminogen activator (tPA). The mean weights of thrombus were [mg (SD)]: Control (saline; n = 8) 39.0 (8.73), tPA 0.01 mg/ml (n = 6) 45.5 (10.56) and tPA 1 mg/ml (n = 8) 3.5 (3.4). Comparing 1 mg/ml tPA vs. 0.01 mg tPA vs. control, p < 0.001 (Mann-Whitney test).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241366 TI - Endothelin-1-like immunoreactivity in postobstructive pulmonary vasculopathy. AB - Postobstructive pulmonary vasculopathy (POPV) is produced by chronic ligation of one pulmonary artery and results in bronchial collateral vessel proliferation and pulmonary arterial abnormalities. The role of endothelin-1 (ET-1), a potent vasoconstrictor peptide, was investigated in a canine model of POPV using radioimmunoassay and immunohistochemistry. The left main pulmonary artery of 9 dogs was ligated, and 3 months (n = 3) and 15 months (n = 6) later, ET-1 levels were measured by radioimmunoassay of plasma samples from left and right pulmonary arteries and veins. In addition, tissues from control and ligated lungs were fixed in Bouin's solution embedded in paraffin and stained with antiserum to ET 1. Plasma ET-1 levels distal to the ligation were not different from those of the control pulmonary artery (1.7 pg/ml in the ligated lungs vs. 1.4 pg/ml in the controls; NS). ET-1-like immunoreactivity was localized mainly to the epithelium in both control and ligated lungs, but the pulmonary arteries and the new bronchial vessels stained more intensely in the ligated lungs. The sections immunostained with ET-1 antiserum preabsorbed with synthetic ET-1 did not stain. These results suggest that ET-1 may play a role in the bronchial neovascularization and the pulmonary arterial thickening characteristic of POPV. PMID- 8241367 TI - Estimating treatment means in a mixed-effect ANOVA model for bioequivalence studies. AB - In order to establish bioequivalence between two formulations in a crossover trial, it is common to assume a mixed-effect analysis of variance (ANOVA) model and perform two one-sided tests. When the analysis is done on the untransformed data, the numerators of the test statistics are not, in general, treatment contrasts. Consequently, the standard errors of the numerators are difficult to compute. The usual practice is to approximate these with the standard errors of treatment contrasts (the "usual approximation"). This paper examines the goodness of this approximation. We present a few technical issues involved in analyzing the untransformed data with a mixed-effect ANOVA model, and state a parametric definition for the terminology "treatment means." The best linear unbiased estimator (BLUE) for the treatment means is derived, as well as its covariance matrix. Due to the presence of the intersubject variability, the variances and covariances of the BLUE of the treatment means are much larger than is commonly believed. A simulation study shows that these larger-than-expected variances/covariances may widen the usual approximate 90% confidence interval by as much as 10%. PMID- 8241368 TI - Proportional hazards model for repeated measures with monotonic ordinal response. AB - In this paper we present a proportional hazards model to determine the effect of predictors on repeated measures data with ordinal response that is monotonic over time. The model can handle both categorical and continuous predictors. It naturally models the time spent in the response states. The assumption of proportional hazards can be partially assessed if replicates (individuals with identical covariate vectors) are available. Censoring can be incorporated into the model, although it does require additional modelling if the censoring is informative. PMID- 8241369 TI - A proportional hazards model for truncated AIDS data. AB - An important source of information on the latency period for AIDS is the data from individuals infected by contaminated blood transfusion. However, a difficulty in the analysis and interpretation of these data is that information is available only on individuals who are infected and develop the disease prior to some specific time. In this paper, we propose an approach to the analysis of such data under a proportional hazards model. The proposed approach allows testing for group effects in the presence of multiple explanatory variables. PMID- 8241370 TI - Group sequential tests for bivariate response: interim analyses of clinical trials with both efficacy and safety endpoints. AB - We describe group sequential tests for a bivariate response. The tests are defined in terms of the two response components jointly, rather than through a single summary statistic. Such methods are appropriate when the two responses concern different aspects of a treatment; for example, one might wish to show that a new treatment is both as effective and as safe as the current standard. We present a formulation of the bivariate testing problem, introduce group sequential tests that satisfy Type I error conditions, and show how to find the sample size guaranteeing a specified power. We describe how properties of group sequential tests for bivariate normal observations can be computed by numerical integration. PMID- 8241371 TI - A multiple-step selection procedure with sequential protection of preferred treatments. AB - Dunnett (1955, Journal of the American Statistical Association 50, 1096-1121) proposed a one-step many-one test to compare several experimental treatment means with a control. However, after the control treatment is rejected, one would want to make comparisons among the experimental treatments. We assume that there is an order of decreasing preference among the experimental treatments based on other considerations. We propose in this paper a multiple-step selection procedure, an extension of Dunnett's one-step many-one test that will give protection to the preferred treatments and guarantee a high selection probability of the correct treatment when that selection is appropriate. We also provide methods to calculate the sample size required to carry out this test for normal, binomial, and exponential data with random censoring. PMID- 8241372 TI - Optimization of multistage testing times and critical values in clinical trials. AB - Optimal critical values and times of intermediate testing points are determined to minimize expected sample size in a variety of situations. Two-stage and three stage models are considered for a variety of values of power. We investigate the robustness of the optimality parameters. PMID- 8241373 TI - An evaluation of some methods for fitting dose-response models to quantal response developmental toxicology data. AB - The analysis of quantal-response developmental toxicology data by dose-response modeling is discussed, with emphasis on methods that avoid exact distributional assumptions. These methods (quasi-likelihood, bootstrapping, and jackknifing) are contrasted with analyses based on the beta-binomial distribution. For the resampling procedures, dose-response models are fit under a binomial likelihood. A justification for this choice of estimator in resampling plans is given, based on an extension of the standard results for asymptotic normality and consistency of maximum likelihood estimators. This justification depends only on the true distribution of the data having the usual binomial expectation. A quasi likelihood approach is also considered, in which simple assumptions about the intralitter correlation structure are made. Quasi-likelihood methods are in theory asymptotically robust to misspecification of the intralitter correlation structure. The practical implications of these asymptotic results are evaluated in a simulation study. PMID- 8241374 TI - Ratio estimates, the delta method, and quantal response tests for increased carcinogenicity. AB - This paper demonstrates the use of the delta method for estimating the variance of ratio statistics derived from animal carcinogenicity experiments. The Cochran Armitage test (Cochran, 1954, Biometrika 10, 417-451; and Armitage, 1955, Biometrics 11, 375-386) is routinely applied to carcinogenicity data as a test for linear trend in lifetime tumor incidence rates. The computing formula for this test derives from the assumption that the denominators of the quantal response rates are fixed. However, when time-at-risk weights are introduced to correct for treatment-related differences in survival, the denominators of the quantal response rates are subject to random variation. The delta method and weighted least squares techniques are applied here to approximate the variance of such ratio statistics and test for a linear dose-response relationship among treatments. This technique is compared to that of Bailer and Portier (1988, Biometrics 44, 417-431), who introduced a survival-adjusted quantal response test for trend in lifetime tumor incidence rates. Their test modifies the usual Cochran-Armitage computing formula by weighting the denominators of the response rates to reflect less-than-whole-animal contributions to risk. Within the framework of a weighted least squares linear regression model that underlies the Cochran-Armitage test, the time-at-risk weights of Bailer and Portier are incorporated using the delta method. Although the delta method approach is slightly more computationally intensive, small-sample simulations indicate that it has superior operating characteristics over the Poly-3 trend test of Bailer and Portier when background tumor incidence rates are low (under 3%) and survival patterns differ markedly across treatments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241375 TI - Maximum likelihood estimation of the attributable fraction from logistic models. AB - Bruzzi et al. (1985, American Journal of Epidemiology 122, 904-914) provided a general logistic-model-based estimator of the attributable fraction for case control data, and Benichou and Gail (1990, Biometrics 46, 991-1003) gave an implicit-delta-method variance formula for this estimator. The Bruzzi et al. estimator is not, however, the maximum likelihood estimator (MLE) based on the model, as it uses the model only to construct the relative risk estimates, and not the covariate-distribution estimate. We here provide maximum likelihood estimators for the attributable fraction in cohort and case-control studies, and their asymptotic variances. The case-control estimator generalizes the estimator of Drescher and Schill (1991, Biometrics 47, 1247-1256). We also present a limited simulation study which confirms earlier work that better small-sample performance is obtained when the confidence interval is centered on the log transformed point estimator rather than the original point estimator. PMID- 8241376 TI - A log-rank test for equivalence of two survivor functions. AB - We consider a hypothesis testing problem in which the alternative states that the vertical distance between the underlying survivor functions nowhere exceeds some prespecified bound delta > 0. Under the assumption of proportional hazards, this hypothesis is shown to be (logically) equivalent to the statement [beta[ < log(1 + epsilon), where beta denotes the regression coefficient associated with the treatment group indicator, and epsilon is a simple strictly increasing function of delta. The testing procedure proposed consists of carrying out in terms of beta (i.e., the standard Cox likelihood estimator of beta) the uniformly most powerful level alpha test for a suitable interval hypothesis about the mean of a Gaussian distribution with fixed variance. The computation of the critical constant of this test is very easy in practice since it admits a representation as the root of the alpha th quantile of a noncentral chi-square distribution with a single degree of freedom. PMID- 8241377 TI - Where is the likelihood ratio test powerful for detecting two component normal mixtures? AB - We compare the power of the likelihood ratio test, the Engelman-Hartigan test, two outlier tests, four goodness-of-fit tests, and eight tests of normality to detect a mixture consisting of two components that are normally distributed with different means but equal variances. We consider the entire range of mixing proportions pi, 0 < pi < 1. For pi > .85 or pi < .15, overall Fisher's skewness statistic is best with Filliben's probability plot correlation coefficient test somewhat less powerful. A combined skewness and kurtosis test, the Anderson Darling test, and the likelihood ratio test are also competitive. For .35 < pi < .65, the Engelman-Hartigan test is best. For other mixing proportions, the likelihood ratio test is best. For situations in which the preferred test had power 50% or more, the power of the likelihood ratio test is also above 50% and within 15 percentage points of the preferred test. PMID- 8241378 TI - A measure of tumorigenic potency incorporating dose-response shape. AB - Many researchers have considered the problem of ranking chemical agents based on their carcinogenic potency. Sawyer et al. (1984, Biometrics 40, 27-40) proposed a carcinogenic potency estimate that incorporates both intercurrent mortality and background tumor rates. Since then, many authors have either generalized the method outlined by Sawyer et al. or developed their own method based on a slightly different adjustment for treatment-related changes in survival. None of these methods, however, has estimated the shape of the dose-response curve and incorporated such an estimate in potency estimation. In this manuscript, a measure of tumorigenic potency is proposed that utilizes the estimated shape of the dose-response relationship, in addition to estimated dose effects, in order to rank chemicals on the basis of carcinogenic risk. Comparison of this new measure to that of Sawyer et al. is done using a large database of animal carcinogenicity experiments from the National Cancer Institute and the National Toxicology Program. PMID- 8241379 TI - Application of generalized estimating equations to a study of in vitro radiation sensitivity. AB - We describe an application of the generalized estimating equation (GEE) method (Liang and Zeger, 1986, Biometrika 73, 13-22) for regression analysis of correlated Poisson data from a split-plot design with a small number of experimental units. As an alternative to the use of an arbitrarily chosen working correlation matrix, we demonstrate the use of GEE with a reasonable model for the true covariance structure among repeated observations within individuals. We show that, under such a split-plot design with large clusters, the asymptotic relative efficiency of GEE with simple (independence or exchangeable) working correlation matrices is rather low. We conclude by summarizing issues and needs for further work concerning efficiency of the GEE parameter estimates in practice. PMID- 8241380 TI - A note on the operating characteristics of the modified F test. AB - Brownie, Boos, and Hughes-Oliver (1990, Biometrics 46, 259-266) suggested a modification to the fixed-effects analysis of variance (ANOVA) F test for use in situations where treatments are likely to affect mean response while simultaneously increasing between-subject variability. These authors suggest that the modified test (1) is robust against departures from population normality, (2) is more powerful than its classical counterpart when the expected alternative form is encountered, and (3) will suffer only minor power losses relative to the usual test when alternatives are in the form of simple shifts in location. This study shows that the modified test is not generally robust when sampling is from nonnormal distributions and is likely to produce unacceptably high Type I error inflations when sampling is from heavy-tailed or skewed distributions. Some power results are also provided for the multisample situation to supplement those of Brownie et al., who generated results only for the two-sample case. PMID- 8241381 TI - Ca channel kinetics during the spontaneous heart beat in embryonic chick ventricle cells. AB - The ability of Ca ions to inhibit Ca channels presents one of the most intriguing problems in membrane biophysics. Because of this negative feedback, Ca channels can regulate the current that flows through them. The kinetics of the channels depend on voltage, and, because the voltage controls the current, a strong interaction exists between voltage dependence and Ca dependence. In addition to this interaction, the proximity of pores and the local concentration of ions also determine how effectively the Ca ions influence channel kinetics. The present article proposes a model that incorporates voltage-dependent kinetics, current dependent kinetics, and channel clustering. We have based the model on previous voltage-clamp data and on Ca and Ba action currents measured during the action potential in beating heart cells. In general we observe that great variability exists in channel kinetics from patch to patch: Ba or Ca currents have low or high amplitudes and slow or fast kinetics during essentially the same voltage regime, either applied step-protocols or spontaneous cell action potentials. To explain this variability, we have postulated that Ca channels interact through shared ions. The model we propose expands on our previous model for Ba currents. We use the same voltage-dependent rate constants for the Ca currents that we did for the Ba currents. However, we vary the current-dependent rate constants according to the species of the conducting ion. The model reproduces the main features of our data, and we use it to predict Ca channel kinetics under physiological conditions. Preliminary reports of this work have appeared (DeFelice et al., 1991, Biophys. J. 59:551a; Risso et al., 1992, Biophys. J. 61:248a). PMID- 8241382 TI - Epifluorescence microscopic studies of monolayers containing mixtures of dioleoyl and dipalmitoylphosphatidylcholines. AB - Monolayers of dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylcholine (DOPC), and some mixtures of these lipids were investigated using an epifluorescence microscopic surface balance. Monolayers were visualized at 23 +/- 1 degree C through the fluorescence of 1 mol% of two different fluorescent probes, 1-palmitoyl-2-(12-[(7-nitro-2-1,3-benzoxadizole-4- yl)amino]dodecanoyl)phosphatidylcholine (NBD-PC), which partitions into the liquid expanded (LE) or disordered lipid phase and 3,3' dioctadecyloxacarbocyanine perchlorate (DiO-C18), which preferentially associates with the liquid condensed (LC) phase or lipid with ordered chains. LC domains were observed in pure DPPC monolayers at relatively low surface pressures (pi), and these domains grew with increasing surface pressure. Only liquid expanded phase was observed in pure DOPC monolayers up to the point of monolayer collapse. In monolayers containing 29:70:1, 49:50:1, and 69:30:1 (mol/mol/mol) of DPPC:DOPC:probe the domains of LC phase were smaller than those seen in DPPC monolayers at equivalent surface pressures. Quantitative analysis of the visual fields shown by the mixed monolayers showed a distribution of sizes of condensed domains at any given pi. At pi = 30 mN m-1, liquid-expanded, or fluid, regions occupied more than 70% of the total monolayer area in all three mixtures studied, whereas DPPC monolayers were more than 75% condensed or solid at that pressure. For monolayers of DPPC:DOPC:NBD-PC 49:50:1 and 69:30:1 the average domain size and the percentage of the total area covered with LC, or rigid, areas increased to a maximum at pi around 35 mN m-1 followed by a decrease at higher pi. Repetitive compression and expansion of the monolayers containing DPPC:DOPC:NBD PC 49:50:1 at an initial rate of 3.2 A2 molecule-1 s-1 produced monolayers with visual properties consistent with there being a preferential exclusion of the unsaturated lipid from the monolayer. PMID- 8241383 TI - Polarization of fluorescently labeled myosin subfragment-1 fully or partially decorating muscle fibers and myofibrils. AB - Fluorescently labeled myosin heads (S1) were added to muscle fibers and myofibrils at various concentrations. The orientation of the absorption dipole of the dye with respect to the axis of F-actin was calculated from polarization of fluorescence which was measured by a novel method from video images of muscle. In this method light emitted from muscle was split by a birefringent crystal into two nonoverlapping images: the first image was created with light polarized in the direction parallel to muscle axis, and the second image was created with light polarized in the direction perpendicular to muscle axis. Images were recorded by high-sensitivity video camera and polarization was calculated from the relative intensity of both images. The method allows measurement of the fluorescence polarization from single myofibril irrigated with low concentrations of S1 labeled with dye. Orientation was also measured by fluorescence-detected linear dichroism. The orientation was different when muscle was irrigated with high concentration of S1 (molar ratio S1:actin in the I bands equal to 1) then when it was irrigated with low concentration of S1 (molar ratio S1:actin in the I bands equal to 0.32). The results support our earlier proposal that S1 could form two different rigor complexes with F-actin depending on the molar ratio of S1:actin. PMID- 8241384 TI - Induction of B-A transitions of deoxyoligonucleotides by multivalent cations in dilute aqueous solution. AB - Circular dichroism (CD) spectra of d(CCCCGGGG) in the presence of Co(NH3)6(3+) are very similar to spectra of r(CCCCGGGG). In contrast, B-form characteristics are observed for d(CCCCGGGG) in the presence of Na+ and Mg2+, even at high salt concentrations. Spermidine induces modest changes of the CD of d(CCCCGGGG). The NMR chemical shifts of the nonexchangeable protons of d(CCCCGGGG) in the absence and presence of Co(NH3)6(3+) were assigned by proton two-dimensional (2D) NOESY and COSY measurements. The chemical shifts of the GH8 protons of d(CCCCGGGG) move upfield upon titration with Co(NH3)6Cl3. The sums of the sugar H1' coupling constants decrease with added Co(NH3)6Cl3. Cross peak intensities in the 2D proton NOESY spectra show a transformation from B-DNA to A-DNA characteristics upon the addition of Co(NH3)6Cl3. The temperature-dependent 59Co transverse and longitudinal relaxation rates demonstrate that Co(NH3)6(3+) is site-bound to the oligomer. Such localization is not a general feature of Co(NH3)6(3+) binding to oligonucleotides. 59Co NMR relaxation and CD measurements demonstrate chiral discrimination by d(CCCCGGGG) for the two stereoisomers of Co(en)3(3+). Both stereoisomers bind tightly as judged by 59Co NMR, and both cause large (but nonequivalent) changes in the CD of this oligomer. PMID- 8241385 TI - Oxygen and CO binding to triply NO and asymmetric NO/CO hemoglobin hybrids. AB - The bimolecular and geminate CO recombination kinetics have been measured for hemoglobin (Hb) with over 90% of the ligand binding sites occupied by NO. Since Hb(NO)4 with inositol hexaphosphate (IHP) at pH below 7 is thought to take on the low affinity (deoxy) conformation, the goal of the experiments was to determine whether the species IHPHb-(NO)3(CO) also exists in this quaternary structure, which would allow ligand binding studies to tetramers in the deoxy conformation. For samples at pH 6.6 in the presence of IHP, the bimolecular kinetics show only a slow phase with rate 7 x 10(4) M-1 s-1, characteristic of CO binding to deoxy Hb, indicating that the triply NO tetramers are in the deoxy conformation. Unlike Hb(CO)4, the fraction recombination occurring during the geminate phase is low (< 1%) in aqueous solutions, suggesting that the IHPHb(NO)3(CO) hybrid is also essentially in the deoxy conformation. By mixing stock solutions of HbCO and HbNO, the initial exchange of dimers produces asymmetric (alpha NO beta NO/alpha CO beta CO) hybrids. At low pH in the presence of IHP, this hybrid also displays a high bimolecular quantum yield and a large fraction of slow (deoxy-like) CO recombination; the slow bimolecular kinetics show components of equal amplitude with rates 7 and 20 x 10(4) M-1 s-1, probably reflecting the differences in the alpha and beta chains. Samples of symmetric hybrids (a2NOI32Co or a2Co922NO) showed a lower (R-like) bimolecular yield and less slow phase for the CO bimolecular recombination, relative to the asymmetric hybrid or the triply NO species. The slower (T state) bimolecular rate of 7 x 104 M-1 s-1 was observed for CO rebinding to a chain.While oxygen equilibrium studies with 'HPHb(NO)3 were hampered by a high oxidation rate, it was possible to perform experiments with samples equilibrated with a mixed CO/oxygen atmosphere. Photodissociation of CO allows a temporary exposure of the binding sites to oxygen. The results confirm that IHPHb(NO)3 has a low oxygen affinity. PMID- 8241386 TI - Responses of two protein-protein complexes to solvent stress: does water play a role at the interface? AB - We have analyzed the stability of the cytochrome c-cytochrome b5 and cytochrome c cytochrome c oxidase complexes as a function of solvent stress. High concentrations of glycerol were used to displace the two equilibria. Glycerol promotes complex formation between cytochrome c and cytochrome b5 but inhibits that between cytochrome c and cytochrome c oxidase. The results with cytochrome b5 and cytochrome c were expected; the association of this complex is largely entropy driven. Our interpretation is that the cytochrome c-cytochrome b5 complex excludes water. The results with the cytochrome c oxidase and cytochrome c couple were not expected. We interpret them to mean that either glycerol is binding to the oxidase, thereby displacing the cytochrome c, or that water is required at this protein-protein interface. A requirement for substantial quantities of water at the interface of some protein complexes is logical but has been reported only once. PMID- 8241387 TI - Cysteine conformation and sulfhydryl interactions in proteins and viruses. 3. Quantitative measurement of the Raman S-H band intensity and frequency. AB - The bond stretching vibration of the cysteine sulfhydryl (SH) group in a typical protein generates a Raman band in the spectral interval 2500-2600 cm-1, a region devoid of interference from any other fundamental mode of vibration of the protein. The relatively high Raman cross section associated with the S-H stretching vibration, the sensitivity of the vibrational frequency to hydrogen bonding interactions and side chain configurations, and the dependence of the Raman intensity on thiol-thiolate equilibria, combine to make the Raman SH band a potentially valuable marker of protein sulfhydryl interactions and a unique indicator of sulfhydryl participation in thiol-disulfide oxidoreductase activity. In order to exploit Raman spectroscopy for these purposes, accurate and precise measurements of Raman SH band profiles are required. We show here that the required precision and accuracy can be achieved by use of the Raman band corresponding to the stretching vibration of in situ nitrogen gas as a quantitative intensity and frequency standard. The Raman Q-branch center of the N2 band occurs at 2330.7 cm-1. PMID- 8241388 TI - Kinetics of specific and nonspecific adhesion of red blood cells on glass. AB - Fixed spherical human red blood cells suspended in 17% sucrose were allowed to adhere on either clean glass surfaces or glass surfaces preincubated with antibodies specific to a certain blood group antigen. The adhesion experiments were performed in an impinging jet apparatus, in which the cells are subjected to stagnation point flow. The objective of this study was to compare the efficiencies of nonspecific and specific (antigen-antibody mediated) adhesion of red blood cells on glass surfaces. The efficiency was defined as the ratio of the experimental adhesion rate to that calculated based on numerical solutions of the mass transfer equation, taking into account hydrodynamic interactions as well as colloidal forces. The efficiency for nonspecific adhesion was nearly unity at flow rates lower than 85 microliter/s (corresponding to a wall shear rate, Gw, of 30 s-1 at a radial distance of 110 microns from the stagnation point). The values of efficiency dropped at higher flow rates, due to an increase in the tangential force. The critical deposition concentration is found to occur at 120-150 mM NaCl, which is consistent with the theoretically predicted values. At low salt concentrations, the experimental values are higher than the theoretical ones. Similar discrepancies have been found in many colloidal systems. Introducing steric repulsion by adsorbing a layer of albumin molecules on the glass completely prevents nonspecific adhesion at flow rates below 60 microliter/s (Gw congruent to 15 s-1). The efficiency of specific adhesion depends both on the concentration of antibody molecules on the surface and the flow rate. Normal red cells adhere more readily through antigen-antibody bonds than fixed cells. Fixed spherical cells have a higher adhesion efficiency than fixed biconcave ones. PMID- 8241389 TI - Langevin dynamics studies of unsaturated phospholipids in a membrane environment. AB - Computer simulations of three unsaturated phospholipids in a membrane environment have been carried out using Langevin dynamics and a mean-field based on the Marcelja model. The applicability of the mean-field to model unsaturated lipids was judged by comparison to available experimental NMR data. The results show that the mean-field methodology and the parameters developed for saturated lipids are applicable in simulations of unsaturated molecules, indicating that these simulations have good predictive capabilities. Single molecule simulations, each 100 ns in length, of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1 palmitoyl-2-elaidoyl-sn-glycero-3-phosphocholine (PEPC), and 1-palmitoyl-2 isolinoleoyl-sn-glycero-3-phosphocholine (PiLPC) reveal similarities between PEPC and DPPC. The presence of the trans double bond in PEPC has a minimum impact on the structural and dynamic properties of the molecule, which is probably the reason that isolated trans double bonds are rare in biological lipids. POPC exhibits different behavior, especially in the calculated average interchain distances, because of the cis double bond. The position of the two double bonds in PiLPC imparts special properties to the molecule. PMID- 8241390 TI - A computer simulation of free-volume distributions and related structural properties in a model lipid bilayer. AB - A novel combined approach of molecular dynamics (MD) and Monte Carlo simulations is developed to calculate various free-volume distributions as a function of position in a lipid bilayer membrane at 323 K. The model bilayer consists of 2 x 100 chain molecules with each chain molecule having 15 carbon segments and one head group and subject to forces restricting bond stretching, bending, and torsional motions. At a surface density of 30 A2/chain molecule, the probability density of finding effective free volume available to spherical permeants displays a distribution with two exponential components. Both pre-exponential factors, p1 and p2, remain roughly constant in the highly ordered chain region with average values of 0.012 and 0.00039 A-3, respectively, and increase to 0.049 and 0.0067 A-3 at the mid-plane. The first characteristic cavity size V1 is only weakly dependent on position in the bilayer interior with an average value of 3.4 A3, while the second characteristic cavity size V2 varies more dramatically from a plateau value of 12.9 A3 in the highly ordered chain region to 9.0 A3 in the center of the bilayer. The mean cavity shape is described in terms of a probability distribution for the angle at which the test permeant is in contact with one of and does not overlap with anyone of the chain segments in the bilayer. The results show that (a) free volume is elongated in the highly ordered chain region with its long axis normal to the bilayer interface approaching spherical symmetry in the center of the bilayer and (b) small free volume is more elongated than large free volume. The order and conformational structures relevant to the free-volume distributions are also examined. It is found that both overall and internal motions have comparable contributions to local disorder and couple strongly with each other, and the occurrence of kink defects has higher probability than predicted from an independent-transition model. PMID- 8241391 TI - Cross-linking myosin subfragment 1 Cys-697 and Cys-707 modifies ATP and actin binding site interactions. AB - Skeletal muscle myosin is an enzyme that interacts allosterically with MgATP and actin to transduce the chemical energy from ATP hydrolysis into work. By modifying myosin structure, one can change this allosteric interaction and gain insight into its mechanism. Chemical cross-linking with N,N'-p phenylenedimaleimide (pPDM) of Cys-697 to Cys-707 of the myosin-ADP complex eliminates activity and produces a species that resembles myosin with ATP bound (Burke et al., 1976). Nucleotide-free pPDM-modified myosin subfragment 1 (S1) was prepared, and its structural and allosteric properties were investigated by comparing the nucleotide and actin interactions of S1 to those of pPDM-S1. The structural properties of the nucleotide-free pPDM-S1 are different from those of S1 in several respects. pPDM-S1 intrinsic tryptophan fluorescence intensity is reduced 28%, indicating a large increase of an internal quenching reaction (the fluorescence intensity of the related vanadate complex of S1, S1-MgADP-Vi, is reduced by a similar degree). Tryptophan fluorescence anisotropy increases from 0.168 for S1 to 0.192 for pPDM-S1, indicating that the unquenched tryptophan population in pPDM-S1 has reduced local freedom of motion. The actin affinity of pPDM-S1 is over 6,000-fold lower than that of S1, and the absolute value of the product of the net effective electric charges at the acto-S1 interface is reduced from 8.1 esu2 for S1 to 1.6 esu2 for pPDM-S1. In spite of these changes, the structural response of pPDM-S1 to nucleotide and the allosteric communication between its ATP and actin sites remain intact. Compared to pPDM-S1, the fluorescence intensity of pPDM-S1 *MgADP is increased 50%(compared to 8 and 31% increases, respectively, for MgADP and MgATP binding to S1). Compared to acto pPDM-S1, the absolute value of the product of the net effective electric charge at the actin binding interface of acto-pPDM-S1 *MgADP increases 7.3 esu2 (compared to a 0.9 esu2 decrease and an 11.0 esu2 increase, respectively, for MgADP and MgATP binding to acto-Sl).The interaction free energy for the ligands MgADP and actin, is -2.0 kcal/mol for pPDM-S1, compared to -1.2 kcal/mol for unmodified S1. PMID- 8241392 TI - Dimeric-like kinetic cooperativity of the bacteriorhodopsin molecules in purple membranes. AB - The kinetics of the absorption changes accompanying the photocycle of bacteriorhodopsin (BR) strongly depend on the intensity of the exciting short laser pulse. The decrease in the flash intensity dependence of the M kinetics after different extents of bleaching of the purple membranes by hydroxylamine proves the existence of a cooperative interaction between the photocycling BR molecules. The yield of the slow component of the M decay (M(s)) is a quadratic function of the extent of the fraction cycling. The slope of the relative weight of M(s) versus the fraction cycling is 0.5. This slope indicates a dimeric-like cooperative interaction, although the structural units of the purple membranes are the trimers of the BR molecules. For the most probable cooperative mechanism an asymmetric trimeric interaction is suggested, which accounts for the apparently dimeric features. A photocycling molecule may influence only one of its two neighbors in the trimer. From this asymmetric feature a deformative interaction is expected to be the cooperative mechanism, which would be an allosteric regulating mechanism in the purple membrane. PMID- 8241393 TI - Quantitation of membrane receptor distributions by image correlation spectroscopy: concept and application. AB - Measurement of receptor distributions on cell surfaces is one important aspect of understanding the mechanism whereby receptors function. In recent years, scanning fluorescence correlation spectroscopy has emerged as an excellent tool for making quantitative measurements of cluster sizes and densities. However, the measurements are slow and usually require fixed preparations. Moreover, while the precision is good, the accuracy is limited by the relatively small amount of information in each measurement, such that many are required. Here we present a novel extension of the scanning correlation spectroscopy that solves a number of the present problems. The new technique, which we call image correlation spectroscopy, is based on quantitative analysis of confocal scanning laser microscopy images. Since these can be generated in a matter of a second or so, the measurements become more rapid. The image is collected over a large cell area so that more sampling is done, improving the accuracy. The sacrifice is a lower resolution in the sampling, which leads to a lower precision. This compromise of precision in favor of speed and accuracy still provides an enormous advantage for image correlation spectroscopy over scanning correlation spectroscopy. The present work demonstrates the underlying theory, showing how the principles can be applied to measurements on standard fluorescent beads and changes in distribution of receptors for platelet-derived growth factor on human foreskin fibroblasts. PMID- 8241394 TI - Crowding-induced organization of cytoskeletal elements: I. Spontaneous demixing of cytosolic proteins and model filaments to form filament bundles. AB - The theory for the effects of crowding on the behavior of reversibly self assembling solutes is extended to mixtures containing nonassembling solutes. The theory predicts that excluded volume will cause dramatic demixing into domains of long, tightly packed, highly aligned fibers coexisting with an isotropic solution of unaggregated species. It suggests that the bundling of fibers in cells is entropically driven and that accessory binding proteins in the cytoplasm serve to modulate the process rather than create it. PMID- 8241395 TI - Macromolecular diffusion in crowded solutions. AB - The effects of crowding on the self or tracer diffusion of macromolecules in concentrated solutions is an important but difficult problem, for which, so far, there has been no rigorous treatment. Muramatsu and Minton suggested a simple model to calculate the diffusion coefficient of a hard sphere among other hard spheres. In this treatment, scaled particle theory is used to evaluate the probability that the target volume for a step in a random walk is free of any macromolecules. We have improved this approach by using a more appropriate target volume which also allows the calculation to be extended to the diffusion of a hard sphere among hard spherocylinders. We conclude that, to the extent that proteins can be approximated as hard particles, the hindrance of globular proteins by other proteins is reduced when the background proteins aggregate (the more so the greater the decrease in particle surface area), the hindrance due to rod-shaped background particles is reduced slightly if the rod-like particles are aligned, and the anisotropy of the diffusion of soluble proteins among cytoskeletal proteins will normally be small. PMID- 8241396 TI - Barium modulates the gating of batrachotoxin-treated Na+ channels in high ionic strength solutions. AB - Batrachotoxin-activated rat brain Na+ channels were reconstituted in neutral planar phospholipid bilayers in high ionic strength solutions (3 M NaCl). Under these conditions, diffuse surface charges present on the channel protein are screened. Nevertheless, the addition of extracellular and/or intracellular Ba2+ caused the following alterations in the gating of Na+ channels: (a) external (or internal) Ba2+ caused a depolarizing (or hyperpolarizing) voltage shift in the gating curve (open probability versus membrane potential curve) of the channels; (b) In the concentration range of 10-120 mM, extracellular Ba2+ caused a larger voltage shift in the gating curve of Na+ channels than intracellular Ba2+; (c) voltage shifts of the gating curve of Na+ channels as a function of external or internal Ba2+ were fitted with a simple binding isotherm with the following parameters: for internal Ba2+, delta V0.5,max (maximum voltage shift) = -11.5 mV, KD = 64.7 mM; for external Ba2+, delta V0.5,max = 13.5 mV, KD = 25.8 mM; (d) the change in the open probability of the channel caused by extracellular or intracellular Ba2+ is a consequence of alterations in both the opening and closing rate constants. Extracellular and intracellular divalent cations can modify the gating kinetics of Na+ channels by a specific modulatory effect that is independent of diffuse surface potentials. External or internal divalent cations probably bind to specific charges on the Na+ channel glycoprotein that modulate channel gating. PMID- 8241397 TI - Critical temperature for unilamellar vesicle formation in dimyristoylphosphatidylcholine dispersions from specific heat measurements. AB - Using a heat conduction calorimeter with very high resolution (+/- 0.00005 J/degrees C.cm3), we have measured the specific heat CpL between 25 and 35 degrees C of dimyristoylphosphatidylcholine (DMPC) in aqueous dispersions. Previous studies of the temperature dependence of the chemical potential of DMPC in the L alpha phase (lamellar, liquid crystalline) indicated that a dispersion consisting only of unilamellar vesicles forms spontaneously at a critical temperature T* of 29.0 degrees C. Our present measurements show an anomaly in CpL between 28.70 and 29.50 degrees C: the curve for CpL versus T first decreases and then exhibits an inflection point at 28.96 degrees C before it flattens. This anomaly is attributed to the transformation from multilamellar dispersion to unilamellar vesicles at T* = 28.96 degrees C. Two independent properties of the CpL data also indicate T* is a critical point for the formation of unilamellar vesicles: (a) the time to reach equilibrium upon changing temperature increased dramatically between 28.7 and 28.96 degrees C, increasing as (T* - T)-1; at T > T* the dramatic "slowing-down" phenomenon was not observed. This slowing-down near T* is a general characteristic of critical phenomena. (b) The free energy change for the multilamellar-unilamellar transformation was obtained from the CpL T data over this temperature interval and found to be 3.2 J/mol or 0.016 ergs/cm2 of bilayer, in agreement with other estimates of the interaction energy between neutral bilayers. We conclude with a discussion of the implications for membrane bilayer stability of these newly identified dynamic properties of the transformation. PMID- 8241398 TI - Preparation of isolated biomolecules for SFM observations: T4 bacteriophage as a test sample. AB - The T4 bacteriophage has been used to investigate protocols for the preparation of samples for scanning force microscopy in air, in order to obtaining reproducible images. The resolution of images and the distribution of bacteriophages on the substrate depends on the buffer type, its concentration, the surface treatment of substrate, and the method of deposition. The best imaging conditions for the phages require dilution in a volatile buffer at low ionic strength and adsorption onto hydrophilic surfaces. When imaging with the scanning force microscopy the quality of the images is influenced by the vertical and lateral forces applied on the sample and by the tip geometry. PMID- 8241399 TI - Interactions between H+ and Ca2+ near cardiac L-type calcium channels: evidence for independent channel-associated binding sites. AB - Monovalent and divalent ions are known to affect voltage-gated ion channels by the screening of, and/or binding to, negative charges located on the surface of cell membranes within the vicinity of the channel protein. In this investigation, we studied gating shifts of cardiac L-type calcium channels induced by extracellular H+ and Ca2+ to determine whether these cations interact at independent or competitive binding sites. At constant pHo (7.4), Cao-induced gating shifts begin to approach a maximum value (approximately equal to 17 mV) at concentrations of extracellular calcium of > or = 40 mM. A fraction of the calcium-dependent gating shift could be titrated with an effective pKa = 6.9 indicating common and competitive access to H+ and Ca2+ ions for at least one binding site. However, if pHo is lowered when Cao is > or = 40 mM, additional shifts in gating are measured, suggesting a subpopulation of sites to which Ca2+ and H+ bind independently. The interdependence of L-channel gating shifts and Cao and pHo was well described by the predictions of surface potential theory in which two sets of binding sites are postulated; site 1 (pKa = 5.5) is accessible only to H+ ions and site 2 (pKa = 6.9) is accessible to both Ca2+ and H+ ions. Theoretical computations generated with this model are consistent with previously determined data, in which interactions between these two cations were not studied, in addition to the present experiments in which interactions were systematically probed. PMID- 8241400 TI - Slow and incomplete inactivations of voltage-gated channels dominate encoding in synthetic neurons. AB - Electrically excitable channels were expressed in Chinese hamster ovary cells using a vaccinia virus vector system. In cells expressing rat brain IIA Na+ channels only, brief pulses (< 1 ms) of depolarizing current resulted in action potentials with a prolonged (0.5-3 s) depolarizing plateau; this plateau was caused by slow and incomplete Na+ channel inactivation. In cells expressing both Na+ and Drosophila Shaker H4 transient K+ channels, there were neuron-like action potentials. In cells with appropriate Na+/K+ current ratios, maintaining stimulation produced repetitive firing over a 10-fold range of frequencies but eventually led to "lock-up" of the potential at a positive value after several seconds of stimulation. The latter effect was due primarily to slow inactivation of the K+ currents. Numerical simulations of modified Hodgkin-Huxley equations describing these currents, using parameters from voltage-clamp kinetics studied in the same cells, accounted for most features of the voltage trajectories. The present study shows that insights into the mechanisms for generating action potentials and trains of action potentials in real excitable cells can be obtained from the analysis of synthetic excitable cells that express a controlled repertoire of ion channels. PMID- 8241401 TI - An approximate model and empirical energy function for solute interactions with a water-phosphatidylcholine interface. AB - An empirical model of a liquid crystalline (L alpha phase) phosphatidylcholine (PC) bilayer interface is presented along with a function which calculates the position-dependent energy of associated solutes. The model approximates the interface as a gradual two-step transition, the first step being from an aqueous phase to a phase of reduced polarity, but which maintains a high enough concentration of water and/or polar head group moieties to satisfy the hydrogen bond-forming potential of the solute. The second transition is from the hydrogen bonding/low polarity region to an effectively anhydrous hydrocarbon phase. The "interfacial energies" of solutes within this variable medium are calculated based upon atomic positions and atomic parameters describing general polarity and hydrogen bond donor/acceptor propensities. This function was tested for its ability to reproduce experimental water-solvent partitioning energies and water bilayer partitioning data. In both cases, the experimental data was reproduced fairly well. Energy minimizations carried out on beta-hexyl glucopyranoside led to identification of a global minimum for the interface-associated glycolipid which exhibited glycosidic torsion angles in agreement with prior results (Hare, B.J., K.P. Howard, and J.H. Prestegard. 1993. Biophys. J. 64:392-398). Molecular dynamics simulations carried out upon this same molecule within the simulated interface led to results which were consistent with a number of experimentally based conclusions from previous work, but failed to quantitatively reproduce an available NMR quadrupolar/dipolar coupling data set (Sanders, C.R., and J.H. Prestegard. 1991. J. Am. Chem. Soc. 113:1987-1996). The proposed model and functions are readily incorporated into computational energy modeling algorithms and may prove useful in future studies of membrane-associated molecules. PMID- 8241402 TI - Mechanism of the electric response of lipid bilayers to bitter substances. AB - In order to clarify by what mechanism the lipid bilayer membrane changes its potential under the stimulation of bitter substances, a microscopic model for the effects of the substances on the membrane is presented and studied theoretically. It is assumed that the substances are adsorbed on the membrane and change the partition coefficients of ions between the membrane and the stimulation solution, the dipole orientation in the polar head, and the diffusion constants of ions in the membrane. It is shown, based on the comparison of the calculated results with the experimental ones, that the response arises mainly from a change in the partition coefficients. Protons play an essential role in the membrane potential variation due to the change in their partition coefficients. The present model reproduces the following observed unique properties in the response of lipid bilayers to bitter substances, which cannot be accounted for by the usual channel model for the membrane potential: 1) the response of the membrane potential appears even under the condition that there is no ion gradient across the membrane, 2) the response remains even when the salt in the stimulating solution is replaced with the salt made of an impermeable cation, and 3) the direction of the polarization of the potential is not reversed, even when the ion gradient across the bilayer is reversed. PMID- 8241403 TI - Thermal equilibration between the M and N intermediates in the photocycle of bacteriorhodopsin. AB - The stages in the photocycle of bacteriorhodopsin (BR) involving the M and N intermediates are investigated using a double pulse excitation method. A first (cycling) pulse at 532 nm is followed, with an appropriate time delay, by a second pulse (337, 406, 446, or 470 nm) which induces the M-->BR back photoreaction. After depletion by the second pulse a repopulation of M in the millisecond range is observed which is interpreted in terms of a thermal N-->M relaxation. It is thus concluded that a (thermal) M<-->N equilibrium accounts for the biphasic decay of M in the BR photocycle. Other models for this stage of the light-driven proton-pump are therefore unnecessary. PMID- 8241404 TI - Histidine substitution identifies a surface position and confers Cs+ selectivity on a K+ pore. AB - The amino acid located at position 369 is a key determinant of the ion conduction pathway or pore of the voltage-gated K+ channels, Kv2.1 and a chimeric channel, CHM, constructed by replacing the pore region of Kv2.1 with that of Kv3.1. To determine the orientation of residue 369 with respect to the aqueous lumen of the pore, the nonpolar Ile at 369 in Kv2.1 was replaced with a basic His. This substitution produced a Cs(+)-selective channel with Cs+:K+ permeability ratio of 4 compared to 0.1 in the wild type. Block by external tetraethylammonium (TEA) was reduced about 20-fold, while block by internal TEA was unaffected. External protons and Zn2+, that are known to interact with the imidazole ring of His, blocked the mutant channel much more effectively than the wild type channel. The blockade by Zn2+ and protons was voltage-independent, and the proton blockade had a pKa of about 6.5, consistent with the pKa for His in solution. The histidyl specific reagent diethylpyrocarbonate produced greatly exaggerated blockade of the mutated channel compared to the wild type. The residue at position 369 appears to form part of the binding site for external TEA and to influence the selectivity for monovalent cations. We suggest that the imidazole side-chain of His369 is exposed to the aqueous lumen at a surface position near the external mouth of the pore. PMID- 8241405 TI - Levels of thermodynamic treatment of biochemical reaction systems. AB - Equilibrium calculations on biochemical reaction systems can be made at three levels. Level 1 is the usual chemical calculation with species at specified temperature and pressure using standard Gibbs energies of formation of species or equilibrium constants K. Level 2 utilizes reactants such as ATP (a sum of species) at specified T, P, pH, and pMg with standard transformed Gibbs energies of formation of reactants or apparent equilibrium constants K'. Calculations at this level can also be made on the enzymatic mechanism for a biochemical reaction. Level 3 utilizes reactants at specified T, P, pH, and pMg, but the equilibrium concentrations of certain reactants are also specified. The fundamental equation of thermodynamics is derived here for Level 3. Equilibrium calculations at this level use standard transformed Gibbs energies of formation of reactants at specified concentrations of certain reactants or apparent equilibrium constants K". Level 3 is useful in calculating equilibrium concentrations of reactants that can be reached in a living cell when some of the reactants are available at steady-state concentrations. Calculations at all three levels are facilitated by the use of conservation matrices and stoichiometric number matrices for systems. Three cases involving glucokinase, glucose-6 phosphatase, and ATPase are discussed. PMID- 8241406 TI - Infrared CD of deoxy oligonucleotides. Conformational studies of 5'd(GCGC)3', 5'd(CGCG)3', 5'd(CCGG)3', and 5'd(GGCC)3' in low and high salt aqueous solution. AB - Infrared (vibrational) circular dichroism (VCD) spectra have been obtained for the self-complementary tetranucleotides, 5'd(CGCG)3', 5'd(GCGC)3', 5'd(CCGG)3', and 5'd(GGCC)3'. In buffered aqueous solution at low salt concentration, these tetramers exhibit spectra associated with right-handed polymers, although the spectra differ significantly for the four species. In high salt solution, a B-->Z transition occurs in 5'd(CGCG)3', while the other tetranucleotides appear only slightly altered. Temperature dependent studies of these oligonucleotides reveal a greater amount of thermal stability for the tetramers in low salt than for the high salt solutions. VCD intensities computed via the exciton formalism are compared with observed results. PMID- 8241407 TI - Cytoplasmic viscosity near the cell plasma membrane: measurement by evanescent field frequency-domain microfluorimetry. AB - The purpose of this study was to determine whether the unique physical milieu just beneath the cell plasma membrane influences the rheology of fluid-phase cytoplasm. Cytoplasmic viscosity was evaluated from the picosecond rotation of the small fluorophore 2',7'-bis-(2-carboxyethyl)-5-carboxyfluorescein (BCECF) by parallel-acquisition Fourier transform microfluorimetry (Fushimi and Verkman, 1991). Information about viscosity within < 200 nm of cell plasma membranes was obtained by selective excitation of fluorophores in an evanescent field created by total internal reflection (TIR) of impulse-modulated s-plane-polarized laser illumination (488 nm) at a glass-aqueous interface. Measurements of fluorescence lifetime and time-resolved anisotropy were carried out in solutions containing fluorescein or BCECF at known viscosities, and monolayers of BCECF-labeled Swiss 3T3 fibroblasts and Madin-Darby canine kidney (MDCK) cells. Specific concerns associated with time-resolved fluorescence measurements in the evanescent field were examined theoretically and/or experimentally, including variations in lifetime due to fluorophore proximity to the interface, and the use of the s and p polarized excitation. In fluorescein solutions excited with s-plane polarized light, there was a 5-10% decrease in fluorescein lifetime with TIR compared to trans (subcritical) illumination, but no change in rotational correlation time (approximately 98 ps/cP). Intracellular BCECF had a single lifetime of 3.7 +/- 0.1 ns near the cell plasma membrane. Apparent fluid-phase viscosity near the cell plasma membrane was 1.1 +/- 0.2 cP (fibroblast) and 1.0 +/- 0.2 cP (MDCK), not significantly different from the viscosity measured in bulk cytoplasm far from the plasma membrane. The results establish the methodology for time-resolved microfluorimetric measurement of polarization in the evanescent field and demonstrate that the cell plasma membrane has little effect on the fluid-phase viscosity of adjacent cytoplasm. PMID- 8241408 TI - Thermodynamics and dynamics of phosphatidylcholine-cholesterol mixed model membranes in the liquid crystalline state: effects of water. AB - A method for obtaining the thermodynamic activity of each membrane component in phosphatidylcholine (PC)/cholesterol mixtures, that is based upon ESR spin labeling is examined. The thermodynamic activity coefficients, gamma PC and gamma chol, for the PC and cholesterol, respectively, are obtained from the measured orientational order parameters, SPC and S(chol), as a function of cholesterol content for a spin-labeled PC and the sterol-type cholestane spin probe (CSL), respectively, and the effects of water concentration are also considered. At water content of 24 weight%, the thermodynamics of DMPC/cholesterol/water mixtures in the liquid-crystalline state may be treated as a two-component solution ignoring the water, but at lower water content the role of water is important, especially at lower cholesterol concentrations. At lower water content (17 wt%), gamma chol decreases with increasing cholesterol content which implies aggregation. However, at higher water content (24 wt%), gamma chol is found initially to increase as a function of cholesterol content before decreasing at higher cholesterol content. This implies a favorable accommodation for the cholesterol in the membrane at high water and low cholesterol content. Good thermodynamic consistency according to the Gibbs-Duhem equation was obtained for gamma PC and gamma chol at 24 wt% water. The availability of gamma chol (and gamma PC) as a function of cholesterol concentration permits the estimate of the boundary for phase separation. The rotational diffusion coefficients of the labeled PC and of CSL were also obtained from the ESR spectra. A previously proposed universal relation for the perpendicular component of the rotational diffusion tensor, R perpendicular, for CSL in PC/cholesterol mixtures (i.e., R perpendicular = R0 perpendicular exp(-AS2chol/RT)) is confirmed. A change in composition of cholesterol or of water for DMPC/cholesterol/water mixtures affects R perpendicular only through the dependence of S(chol) on the composition. In particular, the amount of water affects the membrane fluidity, monitored by R perpendicular for CSL, solely by the structural changes it induces in the membrane for the compositions studied. Rotational diffusion for the labeled PC is found to be more complex, most likely due to the combined action of the internal modes of motion of the flexible chain and of the overall molecular reorientation. PMID- 8241409 TI - Two-dimensional crystallization of Escherichia coli-expressed bacteriorhodopsin and its D96N variant: high resolution structural studies in projection. AB - Highly ordered two-dimensional (2-D) crystals of Escherichia coli-expressed bacteriorhodopsin analog (e-bR) and its D96N variant (e-D96N) reconstituted in Halobacterium halobium lipids have been obtained by starting with the opsin protein purified in the denaturing detergent sodium dodecyl sulfate. These crystals embedded in glucose show electron diffraction in projection to better than 3.0 A at room temperature. This is the first instance that expressed bR or a variant has been crystallized in 2-D arrays showing such high order. The crystal lattice is homologous to that in wild-type bR (w-bR) in purple membranes (PM) and permit high resolution analyses of the structure of the functionally impaired D96N variant. The e-bR crystal is isomorphous to that in PM with an overall averaged fractional change of 12.7% (26-3.6-A resolution) in the projection structure factors. The projection difference Fourier map e-bR-PM at 3.6-A resolution indicates small conformational changes equivalent to movement of approximately < 7 C-atoms distributed within and in the neighborhood of the protein envelope. This result shows that relative to w-bR there are no global structural rearrangements in e-bR at this 3.6 A resolution level. The e-D96N crystal is isomorphous to the e-bR crystal with a smaller (9.2%) overall averaged fractional change in the structure factors. The significant structural differences between e-D96N and e-bR are concentrated at high resolution (5-3.6 A); however, these changes are small as quantified from the 3.6 A resolution e D96N-e-bR Fourier difference map. The difference map showed no statistically significant peaks or valleys within 5 A in projection from the site of D96 substitution on helix C. Elsewhere within the protein envelope the integrated measure of peaks or valleys was < approximately 3 C-atom equivalents. Thus, our results show that for the isosteric substitution of Asp96 by Asn, the molecular conformation of bR in its ground state is essentially unaltered. Therefore, the known effect of D96N on the slowed M412 decay is not due to ground-state structural perturbations. PMID- 8241410 TI - Multiwavelength analysis of the kinetics of reduction of cytochrome aa3 by cytochrome c. AB - Some new approaches to the kinetic study of the reduction of cytochrome aa3 by cytochrome c are presented. The primary innovations are the use of a spectrometer which can acquire multiwavelength data as fast as every 10 microseconds, and the application of a variety of analytical methods which can utilize simultaneously all of the time-resolved spectral data. These techniques include singular value decomposition (SVD), deconvolutions based on pure Gaussian models for absorption peaks, deconvolutions based on isolated absorption spectra for the pure components, and simulations of SVD-deduced and actual experimental difference spectra. The reduction characteristics of the anaerobic resting enzyme can be distinguished from those of pulsed forms. In the former case, only two electrons can be bound by cytochrome aa3, whereas in the latter case complete reduction of the enzyme is achieved. PMID- 8241411 TI - Interaction forces between red cells agglutinated by antibody. IV. Time and force dependence of break-up. AB - We report on an extension of a previously described method to measure the hydrodynamic force to separate doublets of fixed, sphered and swollen red cells cross-linked by antibody (S. P. Tha, J. Shuster, and H. L. Goldsmith. 1986. Biophys. J. 50:1117-1126). With a traveling microtube apparatus, doublets are tracked and videotaped in a slowly accelerating Poiseuille flow in 150-microns diameter tubes, and the hydrodynamic normal force at break-up, Fn, is computed from the measured doublet velocity and radial position. Previous results showed a large range of Fn, the mean of which increased with [antiserum], and an absence of clustering at discrete values of Fn. Since it was assumed that the cells separate the instant a critical force to break all crossbridges was reached, lack of clustering could have been due to the use of a polyclonal antiserum. We therefore studied the effect of monoclonal IgM or IgA antibody on the distribution of Fn. The results showed that the data are as scattered as ever, with Fn varying from 2 to 200 pN, and exhibit no evidence of clustering. However, the scatter in Fn could be due to the stochastic nature of intercellular bonds (E. Evans, D. Berk, and A. Leung. 1991a. Biophys. J. 59:838-848). We therefore studied the force dependence of the time to break-up under constant shear stress (Fn from 30 to 200 pN), both in Poiseuille and Couette flow, the latter by using a counter-rotating cone and plate rheoscope. When 280 doublets were rapidly accelerated in the traveling microtube and then allowed to coast in steady flow for up to 180 s, 91% survived into the constant force region; 16% of these broke up after time intervals, tP, of 2-30s. Of 340 doublets immediately exposed to constant shear in the rheoscope, 37% broke after time intervals, tc, from < 1 to 10 s. Thus, doublets do indeed break up under a constant shear stress, if given time. The average time to break-up decreased significantly with increasing force, while the fraction of doublets broken up increased. At a given Fn, the fraction of break-ups decreased with increasing [IgM], suggesting that the average number of bonds had also increased. Using a stochastic model of break-up (G. I. Bell. 1978. Science (Washington DC). 200:618-627; E. Evans, D. Berk,and A. Leung. 1991. Biophys. J. 59:838-848) and a Poisson distribution for the number of bonds, Nb, break-up in slowly accelerating Poiseuille flow and in immediate shear application in Couette flow was simulated. In Poiseuille flow, the observed range and scatter in Fn could be reproduced assuming (Nb) > 5. In the rheoscope, the time intervals and number of rotations to break-up, tc, were quite well reproduced assuming (Nb) = 4. The similarity of (Fn) for monoclonal IgM and IgA for doublet break-up under constant slow acceleration is compatible with the conclusion of Evans et al. (1991 a) for normal red cells and Xia et al. (manuscript submitted for publication) for sphered and swollen red cells, that the applied force extracts the antigen from the cell membrane. PMID- 8241412 TI - Diffusion coefficient of cyclic GMP in salamander rod outer segments estimated with two fluorescent probes. AB - Experiments have demonstrated that single photoisomerizations in amphibian and primate rods can cause the suppression of 3-5% of the dark circulating current at the response peak (Baylor, D. A., T. D. Lamb, and K. W. Yau. 1979. J. Physiol. (Lond.). 288:613-634; Baylor, D. A., B. J. Nunn, and J. L. Schnapf. 1984. J. Physiol. (Lond.). 357:575-607). These results indicate that the change in [cGMP] effected by a single isomerization must spread longitudinally over at least the corresponding fractional length of the outer segment. The effective longitudinal diffusion coefficient, Dx, of cGMP is thus an important determinant of rod sensitivity. We report here measurements of the effective longitudinal diffusion coefficients, Dx, of two fluorescently labeled molecules: 5/6-carboxyfluorescein and 8-(fluoresceinyl)thioguanosine 3',5'-cyclic monophosphate, introduced into detached outer segments via whole-cell patch electrodes. For these compounds, the average time for equilibration of the entire outer segment with the patch pipette was approximately 6 min. Fluorescence images of rods were analyzed with a one dimensional diffusion model that included limitations on transfer between the electrode and outer segment and the effects of intracellular binding of the dyes. The analyses yielded estimates of Dx of 1.9 and 1.0 microns 2.s-1 for the two dyes. It is shown that these results place an upper limit on Dx for cGMP of 11 microns2.s-1. The actual value of Dx for cGMP in the rod will depend on the degree of intracellular binding of cGMP. Estimates of the effective buffering power for cGMP in the rod at rest range from two to six (Lamb and Pugh, 1992; Cote and Brunnock, 1993). When combined with these estimates, our results predict that for cGMP itself, Dx falls within the range of 1.4-5.5 microns 2.s-1. PMID- 8241413 TI - Evidence for a gamma-turn motif in antifreeze glycopeptides. AB - Knowledge of the secondary structure of antifreeze peptides (AFPs) and glycopeptides (AFGPs) is crucial to understanding the mechanism by which these molecules inhibit ice crystal growth. A polyproline type II helix is perhaps the most widely accepted conformation for active AFGPs; however, random coil and alpha-helix conformations have also been proposed. In this report we present vibrational spectroscopic evidence that the conformation of AFGPs in solution is not random, not alpha-helical, and not polyproline type II. Comparison of AFGP amide vibrational frequencies with those observed and calculated for beta and gamma-turns in other peptides strongly suggests that AFGPs contain substantial turn structure. Computer-generated molecular models were utilized to compare gamma-turn, beta-turn, and polyproline II structures. The gamma-turn motif is consistent with observed amide frequencies and results in a molecule with planar symmetry with respect to the disaccharides. This intriguing conformation may provide new insight into the unusual properties of AFGPs. PMID- 8241414 TI - Colloidal gold particles as an incompressible atomic force microscope imaging standard for assessing the compressibility of biomolecules. AB - Colloidal gold particles have multiple uses as three-dimensional atomic force microscopy imaging standards because they are incompressible, monodisperse, and spherical. The spherical nature of the particles can be exploited to characterize scanning tip geometry. As uniform spheres, colloidal gold particles may be used to calibrate the vertical dimensions of atomic force microscopy at the nanometer level. The monodisperse and incompressible nature of the gold can be used to characterize the vertical dimensions of coadsorbed biomolecules. Simultaneous measurements of gold with tobacco mosaic virus show that, at the same applied vertical force, the tobacco mosaic virus is undamaged by blunt tips but is compressed or disintegrated under sharper scanning styli, suggesting that specimen degradation is partly a pressure-dependent effect. PMID- 8241415 TI - Molecular electrostatic potentials from crystal diffraction: the neurotransmitter gamma-aminobutyric acid. AB - Given the electronic charge parameters obtained from a diffraction study of the charge density distribution in a crystal, a mathematical procedure is presented for deriving the electrostatic potential. The procedure allows the mapping of electrostatic potential for a molecule or group of molecules removed from the crystal structure but with each molecule retaining the effects of polarization owing to the original crystal environment. The method is applied for the neurotransmitter gamma-aminobutyric acid. The potential for a gamma-aminobutyric acid molecule is analyzed in terms of a simple model that is suitable for rapid computations concerned with Coulombic molecular interactions. Outside the molecular envelope at 1.2 A from the atomic nuclei, the total potential is well represented by a sum of spherical atom contributions with V(r) = (q/r) + exp( beta r2). The most important aspherical component in the potential is the dipole contribution from the hydrogen atoms. This can be represented as V(r, phi) = (0.162 cos phi)/(r2 + 0.615). Here, V is in e/A, r is the distance from each nucleus in A, q is the experimentally determined net atomic charge in electron units, and phi is the angle between r and the bond X-H. We obtain beta = 1.47, 1.66, 1.83 A-2 for C, N, and O respectively. For H, no term in beta is needed. PMID- 8241416 TI - Picosecond fluorescence decay of lens protein gamma-II crystallin. AB - The fluorescence decay of tryptophan residues in the bovine lens protein gamma-II crystallin has been measured in aqueous buffer solutions. Results were obtained as a function of emission wavelength, temperature, dissolved oxygen, and denaturing solvent. The protein displayed complex fluorescence decay which fit a biexponential model with a long component (ns) and a short component (few hundred ps). Measured fluorescence quantum yields data for gamma-II crystallin allowed calculation of radiative and non-radiative rate constants. The radiative rate constant was consistent with that observed in other indole derivatives, while the non-radiative rate constant was quite large and accounted for the short lifetime in gamma-II. The temperature dependence of the non-radiative decay in gamma-II crystallin yielded a small activation energy of only 1-2 kcal/mol, compared to 4 kcal/mol for the reference compound NATA whose barrier is known to derive from the rotamer model. PMID- 8241417 TI - Calorimetric determination of the standard transformed enthalpy of a biochemical reaction at specified pH and pMg. AB - In a biochemical reaction there is generally a change in the binding of hydrogen ions and metal ions. Therefore, calorimetric measurements of enthalpies of reaction have to be adjusted for the enthalpies of reaction of the hydrogen ions and metal ions produced or consumed with the buffer. It can be shown that this yields the standard transformed enthalpy of reaction that determines the change in the apparent equilibrium constant K' (written in terms of sums of concentrations of species of a reactant) with temperature at the chosen pH and concentration of free metal ion. The derivations are based on the assumption that the changes in pH and free metal ion concentrations in the calorimetric experiment are small. This assumption is experimentally realized if a solution is well buffered for hydrogen and metal ions. The derived equations are discussed in terms of the implications they have for the performance and interpretation of calorimetric measurements. PMID- 8241418 TI - Thermodynamic parameters of cooperative helix-to-coil transitions from synthetic A-U-rich oligoribonucleotides up to fourteen basepairs. AB - Heat induced helix-to-coil transitions are studied in the form of ultraviolet hypochromicity profiles by absorbance spectroscopy, and delta Cp-curves by differential scanning calorimetry of self-complementary ribonucleotides. The results are analyzed and compared. Van 't Hoff transition enthalpies derived by UV-experiments incorporating concentration variations are found to differ from six-parameter and two-parameter Marquardt-fits on the melting profiles. A measure for the maximum number of nucleotides in intermediate states is obtained from a statistical deconvolution. It yields a range from 12.5% for the shortest nucleotide up to 31.5% for r(UA)7. Model independent calorimetric data are reported. A limit for intra-molecular loop-formation preference is reached by rG(UA)6C within this sequence. PMID- 8241419 TI - Association of nucleosides and their 5'-monophosphates with a tryptophan containing tripeptide, Trp-Leu-Glu: the source of an overestimation by fluorescence spectroscopy. AB - Association of 7-methylguanosine 5'-monophosphate with a tryptophan containing tripeptide, Trp-Leu-Glu, has been studied by fluorescence titration using two different geometries of detection, viz. right angle and front surface geometry. The applicability of these two techniques to determine the stability constant of the nucleotide-peptide adduct is discussed. Evidence is presented that fluorescence titration based on right angle detection may lead to considerable overestimation of the strength of interaction. PMID- 8241420 TI - Theory for two-photon excitation in pattern photobleaching with evanescent illumination. AB - Although the translational diffusion coefficients of proteins embedded in, or tightly bound to, natural and artificial cell membranes have been extensively studied, very little is known about the diffusion of proteins weakly bound to membranes. One method for measuring the diffusion coefficients of weakly bound species is to combine evanescent interference patterns with fluorescence pattern photobleaching recovery (TIR-FPPR). However, this technique is intrinsically limited in that the maximum post-bleach fluorescence change, normalized to the pre-bleach fluorescence, is 0.22. This limitation severely restricts the precision of the data and the applicability of the method. In the theoretical work described herein, it is shown that the maximum fractional fluorescence change in TIR-FPPR with two-photon bleaching and observation would be 0.36. Thus, the use of two-photon excitation in TIR-FPPR would significantly improve the signal-to-noise ratio and would provide more precise measurements of the translational diffusion of proteins weakly bound to surfaces. PMID- 8241421 TI - Berenil recognizes and changes the characteristics of adenine and thymine polynucleotide structures. AB - Using circular dichroism, differential scanning calorimetry and susceptibility to DNAse I cleavage assays, we show that the interaction of berenil, a minor-groove binding drug, with poly(dA-dT).poly(dA-dT) and poly(dA).poly(dT) involves important changes in the polynucleotide conformation. The effect of berenil on poly(dA-dT).poly(dA-dT) comprises a clear alteration in CD spectra even at drug/DNA ratios smaller than the stoichiometric value. Berenil recognizes and binds to the alternating-B conformation of DNA changing it to a new conformation which appears to show some of the peculiarities of poly(dA).poly(dT), possibly through a modification in the helical parameters at the TpA and ApT steps. Such alteration is accompanied by a small calorimetric enthalpy change. Moreover, the calorimetric enthalpy does not change significantly whatever the input ratio of drug to poly(dA-dT).poly(dA-dT), indicating that berenil binding does not substantially alters the enthalpy of transition. In addition to increasing the melting temperature of the polynucleotide, berenil reduces the cooperativity of the poly(dA-dT).poly(dA-dT) transition slightly more than either distamycin or netropsin. PMID- 8241422 TI - Measurements and analyses of electrophoretic mobilities of RAW117 lymphosarcoma cells and their variant cells. AB - The electrophoretic mobilities of the cells of malignant lymphosarcoma cell line RAW117-P and its variant H10 with a highly metastatic property to the liver have been measured at various ionic strengths. The cells of parental cell line (RAW117 P) show higher mobility values in magnitude than those of its variant line (RAW117-H10) in the whole range of electrolyte concentration measured. We have also measured the sialic acid amount carried by cells of both lines. The content of sialic acids in RAW117-H10 cells is observed to be about 27% less than that in RAW117-P cells. The mobility data obtained have been analyzed by a novel mobility formula for colloidal particles with ion-penetrable surface charge layers. The observed mobility difference between RAW117-P cells and RAW117-H10 cells is found to be due to the difference in friction exerted by the cell surface layers on the liquid flow around the cells between these two types of cells and to the difference in fixed-charge density in their surface layers, which is caused by the 27% decrease in sialic acid content. A possible explanation for this mobility difference between these two types of cells is given. PMID- 8241423 TI - Self-association of violamycin (VBI). AB - Violamycin (VBI) self-associates in aqueous solution at concentration greater than 7 x 10(-5) M. We report in this paper visible spectrophotometric and kinetic experiments that characterize this self-association. We started from a simple dimerization model supported by our experimental data, and we used different methods described in literature for the computation of the dimeric extinction coefficient epsilon d at various wavelengths and the dimerization constant Kd = 9300 M-1. PMID- 8241424 TI - Comparative study of the interactions of cisplatin and carboplatin with nucleotides using UV resonance Raman spectroscopy. AB - We have obtained the UV excited resonance Raman spectra of five mononucleotides bound to cisplatin and to carboplatin using excitation in resonance with the first electronic absorption bands of the nucleotide bases. Substantial changes in the spectra are observed following interaction with both platinum drugs, indicating modifications to nucleotide structure. Pt (II) binds to base portions of the nucleotide molecules, altering their normal modes of vibration significantly. We present comparative data of cisplatin and carboplatin, and discuss the implications of these results. The kinetics of the drug/nucleotide reactions differ, but final products are found to be similar. PMID- 8241425 TI - The influence of ionic strength and pH on the aggregation properties of zinc-free insulin studied by static and dynamic laser light scattering. AB - The aggregation properties of zinc-free insulin have been studied using static and dynamic light scattering. The aggregation has been investigated as a function of three parameters, the concentration of sodium chloride (in the range 10-100 mM), the pH value (in the range pH 7.5-10.5), and the insulin concentration (1.8 13.4 mg/mL). The measured homodyne autocorrelation function was used to determine the apparent mean hydrodynamic diameter as well as the apparent weight-averaged molar mass of the insulin species in solution. A method of data analysis was employed, which allows the separation of light scattering contributions from the insulin oligomers and from irrelevant macromolecules and possible impurities present in the sample solutions. Also, a simple phenomenological equilibrium model describing the association of oligomers of insulin is presented. One aspect of this model is that it makes it possible to determine weight average molar masses corrected for virial effects on the Rayleigh ratio. This was necessary because virial effects cannot be isolated and corrected for by dilution since this would change the equilibrium distribution of oligomers. The basis of the model is a positive contribution to Gibbs free energy from charge repulsion depending on the protein charge and the number of monomers in the oligomers, and an assumed constant negative contribution to Gibbs free energy arising from either an entropic gain or hydrogen bonding upon association. The equilibrium model gives a good description of both the apparent weight average molar masses and the apparent hydrodynamic diameters, when the effect of the insulin concentration is taken into account by including virial effects arising from charge-charge repulsion (Donnan effect). The result shows that the association of insulin as a function of pH and ionic strength can be described by an effective charge equal to the charge derived from proton titration reduced by the number of sodium ions binding to insulin. At the lowest pH and highest salt concentration (pH 7.5, 100 mM NaCl, 12 mg/mL insulin), the weight average molar mass is close to that of the hexamer, and at the highest pH and lowest salt concentration (pH 10.5, 10 mM NaCl, 1.9 mg/mL), the weight average molar mass is close to that of the monomer. In all cases, however, a distribution of oligomers is present with a relative Gaussian width of about 30%.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8241426 TI - Refined three-dimensional solution structure of a snake cardiotoxin: analysis of the side-chain organization suggests the existence of a possible phospholipid binding site. AB - The solution structure of toxin gamma (60 residues, 4 disulfides) from Naja nigricollis was determined by proton nmr and molecular modeling with DIANA and X PLOR. The structures were calculated using 489 distance and 81 dihedral angle constraints. The average atomic rms deviation between the nine refined structures and the average structure is 0.118 nm for the backbone atoms. Toxin gamma has an overall folding consisting of three loops stabilized by the four disulfides and forming a two- and a three-stranded beta-sheet (loop I and loops II, III, respectively). The same type of folding has been observed for two homologous cardiotoxins. The very close similarity of the solution structure of toxin gamma and the crystal structure of toxin VII4 includes details of the topological arrangement of numerous side chains. Among these are the conserved residues K12, K18, K35, and Y22, known to be critical for the cytolytic activity of toxin gamma. A cluster of hydrophobic side chains organized around Y22 is found on one side of the three-stranded beta-sheet and is spatially close to a group of three lysines (K12, K18, K35). The side chains of these lysines form a cationic site that can accommodate the binding of a phosphate ion as found in the crystal structure of toxin VII4. The hydrophobic cluster constitutes a possible binding site for the hydrophobic moiety of phospholipids. Together with the complementary cationic site, this hydrophobic surface can form a conserved site by which cardiotoxins bind to membrane phospholipids. PMID- 8241427 TI - Z-->B transition in poly[d(G-m5C)2] induced by interaction with 4',6-diamidino-2 phenylindole. AB - The Z form of poly[d(G-m5C)2], in presence of Mg2+ ion, is found to be transformed into B form upon interaction with 4',6-diamidino-2-phenylindole (DAPI). The Z-->B transformation is complete at a mixing ratio of about 0.07 DAPI per DNA base pairs, i.e., each DAPI molecule may be related to the conversion of 6-7 base pairs. An interaction between DAPI and poly[d(G-m5C)2] in its Z form at low drug: DNA ratios is suggested from optical dichroism and time-resolved luminescence anisotropy results. The spectroscopic behaviour of DAPI indicates that the Z conformation of DNA does not provide normal binding sites for DAPI, such as groove or intercalation sites, but that the initial association may be of external nature. PMID- 8241428 TI - Solid-phase synthesis and stability of triple-helical peptides incorporating native collagen sequences. AB - A generally applicable solid-phase methodology has been developed for the synthesis of triple-helical polypeptides incorporating native collagen sequences. Three nascent peptide chains are C-terminal linked through one N alpha-amino and two N epsilon-amino groups of Lys, while repeating Gly-Pro-Hyp triplets induce triple helicity. Different protecting group strategies, including several three dimensionally orthogonal schemes, have been utilized for the synthesis of four homotrimeric triple-helical polypeptides (THPs) of 79-124 residues, three of which incorporate native type IV collagen sequences. Highly efficient assemblies were achieved by 9-fluorenylmethoxycarbonyl (Fmoc) N alpha-amino group protection, in situ 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate mediated couplings, and 1,8-diazabicyclo [5.4.0] undec-7-ene mediated Fmoc group removal. THPs were characterized by Edman degradation sequencing, size-exclusion chromatography, mass spectrometry, reversed-phase high performance liquid chromatography, and CD spectroscopy. THP thermal stabilities ranged from 35 to 59 degrees C, with chain length and Hyp content being the influential factors. Melting temperatures and van't Hoff enthalpies for peptide triple-helical denaturation could be correlated well to Hyp content. The THP synthetic protocol developed here will allow for the study of both structure and biological activity of specific collagen sequences in homotrimeric and heterotrimeric forms. PMID- 8241429 TI - A and Z canonical conformations in d(CnGCGn) crystals characterized by microFTIR and microRaman spectroscopies. AB - Two crystals d(C2GCG2) and d(C5GCG5) have been studied under microscope by Fourier transform ir spectroscopy and Raman spectroscopy. The x-ray diffraction study of the latter crystal had shown that the d(C5GCG5) sequence is the first DNA dodecamer known to adopt a canonical A conformation [N. Verdaguer, J. Aymami, D. Fernandez-Forner, I. Fita, M. Coll, T. Huynh-Dinh, J. Igolen, and J. A. Subirana (1991) Journal of Molecular Biology, Vol. 221, pp. 623-635]. Characteristic ir marker bands and Raman marker peaks of the A conformation have thus been obtained and are compared with previously proposed assignments correlated to fiber diffraction x-ray results obtained on polymers. The d(C2GCG2) sequence crystal had previously been studied in an intermediate form between B and Z [L. Urpi, J. P. Ridoux, J. Liquier, N. Verdagner, I. Fita, J. A. Subirana, F. Iglesias, T. Huynh-Dinh, J. Igolen, and E. Taillandier (1989) Nucleic Acids Research, Vol. 17, pp. 6669-6679]. In this paper we present results obtained from a crystal with this oligonucleotide in Z conformation. The effect of the crystallization conditions on the geometry of the obtained oligomer helix is discussed. The influence of the addition, to the central tetramer CGCG, of dCn stretches (at the 5' end) and dGn stretches (at the 3' end) of different lengths, on the conformational flexibility of the nucleic acid, is considered. PMID- 8241430 TI - Dynamics and structures of DNA: long-range effects of a 16 base-pair (CG)8 sequence on secondary structure. AB - The effects of inserting 16 base pair (bp) of alternating CG [(CG)8] near the middle of a much longer restriction fragment (1097 bp) are investigated by measuring various properties that are sensitive to secondary and tertiary structure. Results for this fragment are compared with those for a control fragment (1089 bp) with the identical sequence except at the insert. Another fragment (1382 bp), which contains a 296-bp extension at the 5'-end of the 1089 bp control fragment, is also used as a secondary control in some experiments. When the 1097-bp (CG)8 insert fragment is compared with the control fragments in 0.1 M NaCl buffer, the (CG)8 insert is found to induce disproportionately large relative changes in the molar ellipticity at 273 nm ([theta]273), the torsion constant (alpha) measured by fluorescence polarization anisotropy, the optical melting profile, and the susceptibility to S1 nuclease. Estimates of the minimum distance over which the (CG)8 insert alters the secondary structure range from 330 to 550 bp. With increasing NaCl concentration, the 1097-bp insert fragment undergoes a structural transition between 2.0 and 2.5 M that is manifested in the apparent diffusion coefficient (Dplat) from dynamic light scattering at large scattering vector. This transition, which is not exhibited by the control DNAs, is presumed to involve formation of Z-helix at the insert. However, the observed decrease in (Dplat) is attributed to an increase in bending rigidity, which perforce must be globally distributed far beyond the (CG)8 insert per se. In 4.25 M NaCl (but not in 0.1 M NaCl), the addition of 1 ethidium dye per 300 bp induces an extensive structural transition in the 1097 bp (CG)8 insert fragment. This transition, which also is not exhibited by the control DNAs, significantly decreases the bending rigidity, doubles [theta]273, and takes place on a time scale of a few days. Removal of ethidium and salt by dialysis vs 0.1 M NaCl buffer restores the original properties of the 1097-bp (CG)8 insert fragment. The present results are consistent with a (fluctuating, long-range) description of the secondary structure in which a given short sequence transiently fluctuates among two or more distinct secondary structures that extend over much larger domains of variable position and size, and whose relative stabilities depend on distant as well as close-lying base pairs. PMID- 8241432 TI - In-vitro fertilization. PMID- 8241431 TI - Structural changes in 16S RNA from Escherichia coli upon unfolding by urea. AB - The urea-induced unfolding of 16S RNA at low ionic strength has been studied by dynamic light scattering, uv spectroscopy, and some hydrodynamic methods. Three components could be resolved in the photon correlation spectra of scattered light, using the inverse Laplace transform SIPP program [G.R. Danovich and I.N. Serdyuk (1983) in Photon Correlation Techniques in Fluid Mechanics, vol. B38, E.O. Schulz-Dubois, Ed., Springer, Berlin/Heidelberg, New York, p. 315]. One component is assigned to the center-of-mass translation of the RNA, another one to a combination of translational and internal motion, and the last to diffusion of urea clusters. The hydrodynamic dimensions of RNA increase strongly upon transition from 4 to 6 M urea. We conclude that up to 2 M urea, 16S RNA is highly elongated, and coiled above 4 M urea, with a great increase of the hydrodynamic dimensions of RNA being observed upon transition from 4 to 6 M urea. A scheme for RNA unfolding is proposed. PMID- 8241433 TI - The effect of coculture on human zygote development. AB - It is well over a decade since the birth of the first test-tube baby and still the in-vitro conditions for early embryonic development remain suboptimal. The ideal culture medium to increase longevity and improve viability of human embryos is not available. Since the metabolic requirements of the human embryo changes from one cleavage stage to another, the development of a single culture medium for all stages could not be expected. The use of helper cells (coculture) in vitro offers much promise as there are numerous documentations in both man and animals describing their ability to increase blastulation rates and improve embryo viability. This paper reviews the effect of coculture on human zygote development. The selection and establishment of cell-lines, biologic actions of coculture of gametes and zygotes, the outcome, and future prospects are discussed. PMID- 8241434 TI - Micromanipulation of human gametes for assisted fertilization. AB - The techniques of partial zona dissection, subzonal sperm insertion, and intracytoplasmic sperm injection have been clinically employed to alleviate aberrations of human gamete interaction. This article reviews the current status of assisted fertilization technology and its relevance in treating male infertility. Indications for the application of each technique and factors affecting outcome are discussed. PMID- 8241435 TI - Preimplantation diagnosis of genetic disease. AB - Preimplantation diagnosis offers couples at high risk of transmitting a genetic disease the opportunity of prenatal diagnosis before the establishment of pregnancy. The successful application of embryo biopsy techniques, and of the polymerase chain reaction for the amplification of DNA, as well as fluorescent in situ hybridization methods have allowed a small number of IVF units to identify the sex and/or genetic defect in human preimplantation embryos. The development of current methods being applied are reviewed. We suggest that there is a need for constant evaluation of the technique and for improvement of the present methods to ensure reproducibility and accuracy of the diagnostic procedures. PMID- 8241436 TI - Cryopreservation of embryos and oocytes: an update. AB - Widespread incorporation of human embryo cryopreservation into IVF programs may reduce the risk of multiple gestation and severe ovarian hyperstimulation syndrome as well as contributing to an overall increase in pregnancy rates. Slow cooling techniques appropriate to the stage of embryo development are most commonly employed. Growing experience with ultrarapid freezing suggests that this technique may offer similar success rates and has the advantages of simplicity and economy. The developmental stage at cryopreservation has not been conclusively shown to influence successful pregnancy outcome, although several retrospective studies suggest improved embryo survival and pregnancy rate following freezing at the pronuclear stage. Endometrial preparation using a number of different regimens before thawed-embryo transfer appears to confer no advantage over the natural cycle, but may be necessary with ovulatory dysfunction. The use of gonadotropin-releasing hormone (GnRH) analogs during ovarian stimulation for IVF has been suggested to decrease subsequent embryo freeze-thaw survival rates. However, increased pregnancy rates have been reported following transfer of previously cryopreserved embryos originating in cycles using GnRH analog (GnRHa), possibly reflecting increased numbers of embryos available for freezing. Cryopreservation of embryos originating from fertile donor oocytes has been very successful and may be used to facilitate synchronization between a donor and one or more recipients. Recent work in oocyte cryopreservation has addressed the problems of meiotic spindle disruption, the risks of aneuploidy, and decreased fertilization associated with zona hardening and polyspermy. Further refinements in technique will be required before widespread oocyte banking becomes feasible. PMID- 8241437 TI - GIFT, ZIFT, and related techniques. AB - This review focuses on the theoretical backgrounds for tubal gamete and zygote/embryo transfer, as well as the clinical results of gamete intrafallopian transfer (GIFT), which are compared with other non-fertilization procedures in infertile women with patent fallopian tubes. While GIFT and zygote intrafallopian transfer (ZIFT) probably result in a more synchronized entry of embryos into the uterine cavity, prospective, randomized studies have not shown these methods to be preferable to conventional in-vitro fertilization and embryo transfer. Nevertheless, co-culture with various cell types seems to yield more viable embryos with a high rate of implantation. The promising results with co-culture do not seem to be a cell- or species-specific phenomenon. This non-specific positive or negative conditioning effect of co-culture on embryo quality indicates that more optimal culture media for in-vitro fertilization can probably be devised. The requirements of laparoscopy and general anesthesia with GIFT have prompted the development of simpler methods based on fertilization in vivo. Various methods of artificial insemination combined with controlled ovarian hyperstimulation yield comparable results with GIFT in unexplained infertility. In endometriosis, GIFT seems to give better results compared with insemination techniques. Less invasive transcervical gamete and embryo transfer techniques have now been established, obviating the need for operating theater facilities. PMID- 8241438 TI - Legislative efforts affecting the reproductive biology laboratory. AB - Legislative initiatives recently enacted in the USA will affect daily operations in the reproductive biology laboratory. These regulations affect laboratory safety, staffing, quality assurance, and record keeping. They mandate laboratory accreditation including certification, inspection, and proficiency testing. Laboratory standards of practice and costs of operation may be increased with mandatory compliance. PMID- 8241439 TI - Ethical and legal concerns: reproductive technologies 1990-1993. AB - After the rapid increase in reports, bills, and regulations on assisted reproductive technologies (ART) in the 1980s, the first 3 years of the 1990s reveal a continuation of this trend notably in three major aspects. First, a certain consistency has developed in terms of the conditions of accessibility to ART, the definition of infertility, the terms of donation, and the primacy of social filiation, so that anonymity remains controversial. Second, the importance of protection of genetic material has been reaffirmed with regard to gamete and embryo conservation, embryo research, and, in particular, the acceptability of preimplantation diagnosis. Finally, the framework of practices concerning accreditation and control, organization of national data, and management of nominative information has been increasingly refined. PMID- 8241440 TI - Physiology of uterine activity in pregnancy. AB - During the past few years enormous progress has been made in the understanding of the molecular mechanisms involved in parturition; however, the answer to the fundamental question of how labor is initiated remains elusive. This is a very important question because alterations in the timing of birth (preterm and post term deliveries) are associated with much of perinatal morbidity and mortality. Currently available treatments for preterm labor are not clearly effective. Prevention of preterm delivery by home uterine monitoring has been proposed; however, the value of this technique has not been conclusively shown. A variety of substances have been implicated in the genesis of labor, including oxytocin, prostaglandins, cytokines, and endothelin. The role of infection in preterm labor has also been extensively studied, but it seems clear that a relatively small percentage of preterm labor is caused by infection. Attention has also focused on the role of estrogen and progesterone, and the possible uses of progesterone antagonists in the induction of labor. A better understanding of the relationship of intrauterine hypoxia and preterm delivery may also help us in establishing treatment and prevention strategies. PMID- 8241441 TI - Antenatal care and intrapartum management. AB - Cost-effective care is an increasingly important issue in medicine. This will necessitate re-evaluating many aspects of antenatal and intrapartum care, including health-care delivery systems and routine surveillance techniques for uncommon events such as Down syndrome. It will demand justification for broadening indications for cesarean section by including fetal macrosomia and intensify the search for the causes of preterm labor. As new techniques are developed for intrapartum monitoring, such as fetal electrocardiography, fetal lactic acid levels, and near-infrared spectroscopy, they, too, will have to be determined to be cost-effective before attaining a role in clinical management. PMID- 8241442 TI - Regional anesthesia and analgesia in obstetrics. AB - Regional analgesia and anesthesia for obstetrical patients are undergoing revolutionary changes which will, ultimately, benefit parturients and neonates. These changes have taken place in the arena of techniques, equipment, as well as in medications. This review will cover the management techniques both for vaginal delivery and cesarean section. PMID- 8241443 TI - Multiple pregnancy. AB - The incidence of multiple gestations has increased markedly over the past decade. This review will focus on new interventions and techniques such as selective fetal reduction, optimizing maternal weight gain, bed rest, home uterine monitoring, ultrasound, and Doppler velocimetry, which have been employed to improve outcomes in these pregnancies. PMID- 8241444 TI - Breech presentation. AB - This review covers recent literature relevant to breech presentation. Some have advocated routine elective cesarean delivery of the breech fetus of any gestational age to reduce the alleged increase in perinatal morbidity and mortality associated with breech presentation. This indication for cesarean delivery accounts for approximately 15% of all cesarean deliveries and costs society approximately US$ 1.4 billion per year. As a result of concerns with vaginal breech delivery, external cephalic version in the term fetus has become well accepted. A new scoring system has been developed to predict the success of external cephalic version. Repeated external cephalic version has been shown to be successful. Careful ultrasound continues to be important before the procedure. It has been suggested that magnetic resonance pelvimetry may be a practical alternative in evaluating candidates for breech vaginal delivery. There is still no compelling prospective data supporting cesarean delivery for preterm breech presentation. Legal considerations have prevented many from attempting vaginal breech delivery, even in carefully selected patients. PMID- 8241445 TI - Prolonged pregnancy. AB - The perceived risks to the fetus of the postmature syndrome have led to the obstetrician's dilemma of whether to await spontaneous parturition or to artificially bring forward the onset of labor to an arbitrarily defined gestation. A prolonged or post-term pregnancy relates to a gestation of greater than 42 weeks and should not be regarded as synonymous with postmaturity. The latter describes a clinical syndrome thought to be a consequence of failing placental function. It is the difficulty in identifying the 'at risk' fetus which has led to routine induction policies. But is such a policy justified in terms of fetal outcome, and if so, is the mother put at increased risk? This review puts into perspective the most recent publications and points out areas requiring further study. PMID- 8241446 TI - Treatment of obstetrical hemorrhagic emergencies. AB - The common hemorrhagic complications of pregnancy remain among the leading causes of maternal mortality. We review new methods of diagnosis and offer suggestions for management. Abruptio placenta, disseminated intravascular coagulation, placenta previa, uterine rupture, and postpartum hemorrhage are included in this review. PMID- 8241447 TI - Puerperium and breast-feeding. AB - This review focuses on several aspects of breast-feeding, including mothers' skills and attitudes, risk of HIV transmission in breast milk, lactational amenorrhea and its contraceptive effects, and the effects of anesthesia and analgesia on lactation. Also discussed is postpartum contraception and sources of referral for low-income women desiring assistance in nutrition. PMID- 8241449 TI - In-vitro fertilization. PMID- 8241448 TI - 'Routine' antenatal care. AB - The concept of antenatal care is a good model of preventive health targeted towards primary and secondary prevention of diseases and pathological conditions during pregnancy and delivery. A critical evaluation of antenatal care programs demonstrated that these are generally associated with a wide range of positive effects, although the type of care provider is not. However, research methodological limitations such as the selection bias (more care is received by healthier women who will have better outcomes anyway), cannot be ruled out as an explanation for the positive results. PMID- 8241450 TI - General obstetrics. PMID- 8241451 TI - Anticholinergic effects of strychnine in the cochlea do not involve muscarinic receptors. AB - Central control of cochlear function is mediated by the cholinergic (medial) efferent system and both muscarinic and nicotinic acetylcholine receptors are thought to be present on outer hair cells. All the physiological effects of acetylcholine in the cochlea are blocked by strychnine and we therefore investigated whether strychnine interacts with muscarinic receptors in the cochlea. The effects of strychnine on both (3H)-quinuclidinyl benzylate binding and atropine sensitive carbachol-induced (3H)-inositol phosphate formation were examined. Strychnine (1 to 50 microM) has no effect on either quinuclidinyl benzylate binding or carbachol (1 mM)-induced inositol phosphate synthesis. Moreover, strychnine does not change basal inositol phosphate metabolism. These data indicate that muscarinic receptors are not sensitive to strychnine at concentrations which are known to block the effects of acetylcholine on outer hair cells. PMID- 8241452 TI - Inflammation-induced upregulation of NK1 receptor mRNA in dorsal horn neurones. AB - The expression of the neurokinin 1 (NK1) receptor (i.e. substance P receptor) gene in spinal cord was studied in rats subjected to unilateral inflammation by semi-quantitative in situ hybridization analysis. Low levels of NK1 receptor mRNA were detected in many neurones throughout the grey matter. Relatively strong labelling was observed in large motoneurones and a subpopulation of superficial dorsal horn neurones. Six days after Freund's adjuvant-induced unilateral hindpaw inflammation, NK1 receptor mRNA levels in lamina I/II of the dorsal horn ipsilateral to the inflamed paw increased almost two fold compared with the contralateral side. These data suggest an inflammation-induced increase of NK1 receptor synthesis in intrinsic spinal cord neurones involved in nociceptive neurotransmission. PMID- 8241453 TI - Responses of cells in cat's area 17 to random dot patterns: influence of stimulus size. AB - We have investigated both the response of cells in the striate cortex to a moving random dot pattern (visual noise) and the extent to which these responses depend on stimulus size. Cell receptive fields (RF) were classified according to the level of modulation of their response to sinusoidal gratings and the spatial arrangement of their ON and OFF regions. Our results indicate that most complex and simple cells respond to visual noise. The amplitude of these responses depend largely on the area of the visual field that was stimulated. Most cells were optimally activated when the noise covered a region much larger than the RF. Our results suggest that the area beyond the classical RF may contribute to the cells' responsiveness to noise. PMID- 8241454 TI - A new type of interplexiform cell in the goldfish retina is PNMT-immunoreactive. AB - A new cell type immunoreactive for phenylethanolamine N-methyltransferase, the synthesizing enzyme for epinephrine, has been identified in the goldfish retina. The somata of the immunoreactive cells were located in the proximal inner nuclear layer and immunoreactive processes were located in both the inner and outer plexiform layers, suggesting that this cell may be an interplexiform cell. Confocal microscopy was used to establish that the putative adrenergic neurites in the outer plexiform layer were located between the somata of horizontal cells and photoreceptor cell synaptic terminals. Intracellular recordings from horizontal cells demonstrated that epinephrine had an effect on horizontal cells, but that the action of epinephrine was consistent with an effect on dopamine receptors, rather than adrenergic receptors. The function of the putative 'I3' interplexiform cell therefore remains unclear, possibly modulating horizontal cell function with a transmitter other than epinephrine or affecting photoreceptors or the retinal pigment epithelium rather than horizontal cells. PMID- 8241455 TI - Antioestrogen inhibits myelination in brains of juvenile Zebra finches. AB - The effects of the antioestrogen Keoxifene on the ontogenetic process of myelination, on the differentiation of neurones of telencephalic song motor centres, on cerebellar structures and on behaviour were studied in male Zebra finches. Brain differentiation was studied by using computer aided analysis of silver impregnated brain sections and by measuring soma sizes of neurones after Nissl staining. An antioestrogen induced inhibition of myelination could be detected in the song motor centre robust nucleus of the archistriatum (RA) and in the cerebellum, whereas the region magnocellular nucleus of the anterior neostriatum (MAN) showed no difference between treated birds and controls. PMID- 8241456 TI - Expression of p75NGFR in the olfactory system following peripheral deafferentation. AB - Following chemically induced peripheral deafferentation of the main olfactory bulb, expression of the low affinity receptor for neurotrophins, p75NGFR, was monitored immunohistochemically using monoclonal antibody MAb192. In control animals, p75NGFR expression was localized to the glomerular layer of the main olfactory bulb and deepest one third of the olfactory neuroepithelium. Peripheral lesioning dramatically reduced expression of glomerular p75NGFR while inducing expression of p75NGFR in the olfactory nerve layer. Expression of p75NGFR in the lesioned neuroepithelium was disorganized compared with controls. These observations demonstrate that the maintenance of the peripheral input from the neuroepithelium is critical to expression of p75NGFR in the main olfactory bulb. PMID- 8241457 TI - Nitric oxide supersensitivity: a possible molecular mechanism of migraine pain. AB - Nitroglycerin, which may be regarded as a prodrug for nitric oxide, induces a mild to moderate headache in healthy subjects. In order to study whether migraine patients are more sensitive to nitric oxide than non-migrainous subjects, four different doses of intravenous nitroglycerin were given in a double blind design to 17 migraine patients, 17 age and sex matched healthy controls and 9 subjects with tension-type headache. The nitroglycerin-induced headache was significantly more severe in migraine sufferers, lasted longer and fulfilled diagnostic criteria for migraine more often. We have previously shown a similar supersensitivity to histamine which in human cerebral arteries activates endothelial H1 receptors and causes endothelial production of nitric oxide. Migraine patients are thus supersensitive to exogenous nitric oxide from nitroglycerin as well as to endothelially produced nitric oxide. It is suggested that nitric oxide may be partially or completely responsible for migraine pain. PMID- 8241458 TI - Antioxidant protection of bio-normalizer in cerebral ischaemia-reperfusion injury in the gerbil. AB - In this study we used cerebral ischaemia-reperfusion injury (IRI) as an experimental model to analyse the effects of Bio-normalizer (BN, a naturally fermented health food product) on reactive oxygen species related changes in different brain regions of gerbils. Pre-administration of BN solutions (0.1% and 1%) for 45 days produced a significant reduction in IRI-mediated increase in membrane lipid peroxidation, as shown by the decreased carbon-centred radicals and thiobarbituric acid-reactive substances in several brain regions including the cerebral cortex, the hippocampus and the striatum. BN however has no effect on the mitochondrial superoxide dismutase activity. PMID- 8241459 TI - Guanidinoethane sulfate: brain pH alkaline shifter. AB - A new category of agents, brain pH alkaline shifters, is described. Using the prototype agent, guanidinoethane sulfate (GES), the actual alkaline shift in pH was demonstrated in adult mice brain by 31-phosphorus (31P) nuclear magnetic resonance (NMR) in vivo spectroscopy. This alkaline shift was also shown to effectively reduce the extent of brain intracellular lactic acidosis brought about by anoxic insult. These findings support the notion that a pH alkaline shift may protect the brain against the deleterious effects of lactic acidosis. Since higher pH has been shown to significantly reduce beta-amyloid deposition, alkaline shifters may also have therapeutic potential in Alzheimer's disease. PMID- 8241460 TI - Cells with a familial Alzheimer's disease mutation produce authentic beta peptide. AB - Cells overexpressing the beta-amyloid precursor protein possessing a mutation found in familial Alzheimer's disease overproduce beta-amyloid peptide (A beta). Because these findings were based on immunological identification, we have chemically characterized the peptides produced. Purified A beta fragments from the conditioned media of these cells were found to have N-terminal sequence consistent with the A beta found in cerebral plaques. Mass spectrometric data demonstrated a series of A beta fragments consistent with those found in Alzheimer's disease (AD); the major species corresponding to A beta(1-40). Significantly, a longer fragment corresponding to A beta(1-42) was found. These findings suggest that this cellular system may be useful for mechanistic studies of A beta generation and possibly for the development of therapeutic agents to treat AD. PMID- 8241461 TI - In vivo monoamine release during naloxone-precipitated morphine withdrawal. AB - There is much evidence from animal work suggesting that the release of noradrenaline (NA) in the brain increases during naloxone-precipitated morphine withdrawal, but the evidence in favour of changes in release of 5 hydroxytryptamine (5-HT) and dopamine (DA) is contradictory. Here we demonstrate, using in vivo microdialysis, that whilst there is a considerable increase (300%) in release of NA in hippocampus precipitated by naloxone in morphine-dependent rats, there is no change in the release of either 5-HT (in hippocampus) or DA (in nucleus accumbens). These results are consistent with suggestions that the symptoms of morphine withdrawal in rats are due primarily to an increase in central NA release. PMID- 8241462 TI - Interleukin-2 enhances scopolamine-induced amnesia and hyperactivity in the mouse. AB - We studied the effects of human recombinant interleukin-2 (IL-2) on scopolamine induced amnesia for a passive avoidance response and on scopolamine-induced hyperactivity, in the mouse. The pre-training intraperitoneal administration of the cytokine significantly enhanced the amnesic effect of scopolamine (1.0 mg kg 1 i.p.). Similar effects were observed after repeated administration (2,500 IU/mouse i.p. for 10 days) of the cytokine. In this condition, the treatment with the pro-cholinergic drug acetylcarnitine completely blocked the pro-amnesic effect of IL-2. The cytokine did not affect plasma glucose concentrations, nor induced any change in footshock sensitivity. Intraperitoneal injection of IL-2 significantly enhanced the scopolamine-induced hyperactivity, whereas the cytokine alone was ineffective in modifying locomotor activity. Our results suggest an involvement of the cholinergic system in the neuromodulatory action of IL-2. PMID- 8241463 TI - Intracerebroventricular infusion of glycine stimulates vasopressin release in conscious sheep. AB - We have previously observed that elevation of the plasma glycine concentration stimulates vasopressin (AVP) release in man and sheep. In the present study we show that this effect of glycine can be elicited directly via a cerebral site of action in conscious sheep. Intracerebroventricular infusions of 0.05 and 0.15 M glycine solutions increased the plasma AVP concentration by 6 and 50 times, respectively, while no effect on water intake was observed. Hyperhydration did not block the stimulation of AVP release. No behavioural side effects, or changes in blood pressure, were observed in response to the infusions. PMID- 8241464 TI - NH4Cl-induced inward currents and cytoplasmic Ca2+ transients in chick sensory neurones. AB - In neurones cultured from chick dorsal root ganglia, application of NH4Cl (3-45 mM) produced a transient inward current followed by a sustained current at negative holding potentials. Methylamines, hydrazine and guanidine were not able to mimic the effects of NH4Cl. The transient, but not the steady current, was inactivated during successive applications of NH4Cl. Challenge with acidic solutions (pH 6.0-6.4) or Ca(2+)-free solutions induced similar currents and abolished NH4Cl-induced transients. Exposure to 10 mM NH4Cl transiently increased cytoplasmic free Ca2+, which then fell to a sustained plateau. NH4Cl-induced membrane depolarization and the concomitant elevation in intracellular Ca2+ can play an important role in modulation of neuronal activity. PMID- 8241465 TI - Subtle variations in living conditions influence behavioural response to d amphetamine. AB - The influence of prior living conditions on behaviours related to the reinforcing properties of d-amphetamine was determined. Rats were housed either in bright light (BL) conditions at the top of a standard rack or in dim light (DL) conditions on the third row of the same rack. Although BL rats showed a higher locomotor response in the novel environment, they were less sensitive than DL rats to the locomotor activating effects of d-amphetamine. The groups did not differ in responding for a secondary reinforcer in a conditioned reinforcement paradigm. However, only in DL rats as the response for the secondary reinforcer enhanced by d-amphetamine. These findings indicate that subtle differences in housing conditions can influence behavioural responses to psychoactive drugs. PMID- 8241466 TI - NGF is released into plasma during human pregnancy: an oxytocin-mediated response? AB - The presence of biologically active nerve growth factor (NGF) in the peripheral circulation of women during pregnancy, labour and lactation was investigated. Using a sensitive immunoenzymatic assay (ELISA), we found an approximately five fold increase in plasma NGF levels during labour and lactation compared with the concentrations found at the term of gestation or in control healthy women. Since labour and lactation are characterized by activation of the hypothalamo-pituitary adrenal axis and by high plasma levels of the neurohypophyseal hormone oxytocin, and since the intravenous injection of oxytocin in female rats causes a 176% increase in the hypothalamic levels of NGF, it is possible that the increased amount of circulating NGF is correlated with one or both of these events. PMID- 8241467 TI - NADPH-diaphorase activity in the central auditory structures of the rat. AB - Distribution of NADPH-diaphorase (NADPH-d) positive neurones was investigated in the central auditory structures of the rat. NADPH-d positive perikarya were found in the dorsal and external cortices of the inferior colliculus and in the intercollicular commissure. Within the medial geniculate body (MGB), stained cell bodies were observed in the dorsal part of the suprageniculate nucleus and in the basal part of the medial division of the MGB. Occurrence of the NADPH-d positive perikarya in the MGB is limited to its caudal third. NADPH-d positive cells are dispersed in the auditory temporal cortex within layers II-VI, with a slight prevalence in the infragranular layers. NADPH-d positive neurones have smooth dendrites without stained dendritic spines. PMID- 8241468 TI - The RNA editing process in Trypanosoma brucei. AB - Twelve mitochondrial mRNAs are edited in Trypanosoma brucei, nine extensively, by addition and removal of uridines. The accumulation of the edited RNAs is regulated during the life cycle. Hundreds of different gRNAs, encoded three or four per minicircle, specify the editing and minicircle content accounts for variation in editing among species and in mutants. The current understanding of the process of gRNA utilization, the editing mechanism and the editing machinery is discussed. PMID- 8241469 TI - Apolipoprotein B mRNA editing: the sequence to the event. AB - Editing of the mRNA coding for apolipoprotein B involves a cytidine to uridine transition at nucleotide 6666 and enables translation of two protein variants. The development of in vitro editing systems has led to the identification of three sequence requirements in this process. The mechanism for C-->U editing requires specific sequences for editing site recognition, positioning of the catalytic activity and enhancement of catalytic efficiency. The dependence of in vitro editing on extract proteins has focused future research in this field on the identification of factors involved in apoB mRNA editing and the role of these factors in the assembly of ribonucleoprotein editosomes. PMID- 8241470 TI - Double-stranded RNA adenosine deaminase as a potential mammalian RNA editing factor. AB - Double-stranded RNA (dsRNA) adenosine deaminase, or DRADA, is a cellular enzyme that modifies adenosine residues to inosines in dsRNA by hydrolytic deamination, replacing A-U with mismatched I-U base pairs. Since it alters the base composition in its substrate RNA, one possible role played by DRADA is to participate in RNA editing. In this article, a brief review is given of characteristics of DRADA. Its possible involvement in RNA editing is also discussed in detail, including specific cases in which DRADA has been implicated as an RNA editing factor. PMID- 8241471 TI - Time for a crisis. PMID- 8241472 TI - The impact of total quality management on health and safety. PMID- 8241473 TI - Diary of an occupational physician. PMID- 8241474 TI - Occupational health needs in small industry in New Zealand: preferred sources of information. AB - An audit of occupational health needs assessment and service utilization was carried out in a survey of small industry. Fifteen per cent of the workforce considered that working conditions were poor, and 8 per cent considered that their health had been affected by work. The proportions were highest in manual workers, rising to 31 per cent and 18 per cent respectively. Thirty-eight per cent of the workforce had needed occupational health information or advice in the past, particularly amongst manual workers (53 per cent). The 'boss' was the commonest source of information, followed by the general practitioner. These findings have important implications for planners of occupational health services who need to ensure that these two groups have access to high quality information. Fifty-nine per cent of those who had needed information had not actually sought and obtained it. This suggests that information needs are not being met. Information which was received, however, generally met the requester's need. PMID- 8241475 TI - Mixed solvent exposure and hearing impairment: an epidemiological study of 3284 men. The Copenhagen male study. AB - Animal experiments and human studies have indicated an effect on auditory functions from exposure to organic solvents. In this study the relationship between self-assessed hearing problems and occupational exposure to solvents was investigated in a cross-sectional design with 3284 participating men aged 53-74 years. Exposure to solvents for five years or more resulted in an adjusted relative risk (RR) for hearing impairment of 1.4 (95 per cent CI: 1.1-1.9) in men without occupational exposure to noise. Factors adjusted for were age, noise traumas, chronic middle ear infection and family history of hearing impairment. The prevalence of hearing impairment in men not exposed to organic solvents was 24 per cent and the attributable risk from solvent exposure was 9.6 per cent. Exposure for less than five years had no effect on hearing capacity. Occupational exposure to noise for five years or more had an effect twice that of solvents, RR: 1.9 (95 per cent CI: 1.7-2.1). In men exposed to both solvents and noise the effect of the latter dominated and no additional effect from solvents was found. A subsample of 51 men was examined with pure tone audiometry and 20 of 21 men who reported abnormal hearing also fulfilled an audiometric criterion for hearing impairment. In conclusion a damaging effect on hearing ability from long-term solvent exposure was found in the present study. The relative effect was moderate but with a high background frequency of hearing problems in the unexposed sample the absolute effect, ie attributable risk, was considerable and of both clinical and preventive importance. PMID- 8241476 TI - Hearing conservation in the mining industry. Evaluation of a risk factor questionnaire. AB - The objectives of this study were to assess the validity of a self-administered hearing questionnaire in the assessment of risk to hearing, to amend the questionnaire design as necessary, and to give general guidance on the use of audiometric questionnaires in hearing conservation programmes. The study was carried out at a colliery in Staffordshire. The odds of deafness, according to the Health and Safety Executive definition, were analysed by exposure to the hearing risk variables in the questionnaire. The results of the hearing questionnaire were linked to a cross-sectional audiometric survey. There were 189 questionnaires eligible for inclusion. Noise exposure at work was reasonably well predicted by having to shout at work (OR 1.42, 95 per cent CI 0.58-3.54) and being exposed to noisy tools and machinery (OR 2.12, 95 percent CI 0.72-6.72). Risk to hearing from disease was predicted by having 'abnormal' hearing or discharging ears, but not by any of the other exposure variables. Risk to hearing from acoustic trauma was demonstrated for fire-arms (OR 1.23, 95 per cent CI 0.5 2.98). None of the hobbies included in the questionnaire usefully predicted hearing loss. The conclusions are that, contrary to Health and Safety Executive advice, audiometric questionnaires may reasonably be administered to all employees, as they are useful in the documentation of hearing loss risk factors. They should not, however, be used to detect ear disease. A revised questionnaire was drafted as the result of these findings, and will be piloted in a larger population. PMID- 8241477 TI - Pre-employment screening of NHS employees with epilepsy. AB - A survey was carried out on whether district health authorities have adopted clear policies with regard to the pre-employment screening of people with epilepsy. The survey revealed that only a small number have done so and that a few others claimed to be following recommended guidelines. It is evident that there is a need for an organization, such as the Association of National Health Service Occupational Physicians, to develop a more authoritative and comprehensive set of guidelines in this area. The paper concludes by suggesting some of the key elements to be included in such a set of guidelines. PMID- 8241478 TI - Cholinesterase assessment as a result of fenitrothion exposure: a survey in a group of public health workers exposed to an organophosphorus pesticide. AB - Antimalarial spraying activities are very important for the malaria control programme in Central Region, Sudan. Fenitrothion, an organophosphorus pesticide, is used for that purpose. A survey of 17 spraying group workers plus three controls was carried out. Whole blood cholinesterase determination was done using the Tintometric method. The maximum and minimum temperatures recorded on the days of the survey were 99 and 78 degrees F respectively. The relative humidity ranged between 82 and 40 per cent in the morning and afternoon respectively. Cholinesterase levels were measured on two pre-start days and then pre- and post exposure levels were determined on 13 days of a 42 day spraying campaign. On 11 out of the 13 days, cholinesterase levels showed a significant drop. The cholinesterase levels of some members of the spraying team decreased by 25 per cent, 37.5 per cent or 50 per cent which were considered as slight-moderate, moderate-pronounced and pronounced inhibition, respectively. PMID- 8241479 TI - The value of questionnaires and spirometry in asthma surveillance programmes in the workplace. AB - Shipyard workers and job applicants (n = 1126) completed an asthma questionnaire, and underwent measurements of ventilatory lung function (FEV1, FEV1/FVC and PEFR) and airway responsiveness. Airway responsiveness (to inhaled methacholine) was expressed as the dose calculated to provoke a 20 per cent fall in FEV1, ie the PD20. Results were categorized into four levels: high (PD20 < 200 micrograms), medium (200-1000 micrograms), low (1001-6400 micrograms) and unquantifiable (> 6400 micrograms), which correspond with definite, possible, doubtful and no asthmatic activity. These categories agreed closely with diagnoses of asthma by general practitioners and the use of bronchodilator medication. The sensitivity and specificity of the questionnaire and the measurements of ventilatory function for detecting asthmatic activity were then determined. The questionnaire symptoms (wheeze, chest tightness, undue coughing or abnormal breathlessness) had a low (28 per cent) sensitivity for detecting definite or possible asthmatic activity and a specificity of only 73 per cent. The sensitivity of the ventilatory function tests (any one abnormal) was also low at 21 per cent with a specificity of 92 per cent. When the FEV1 < 80 per cent predicted criterion was considered separately, its sensitivity was 11 per cent and its specificity was 98 per cent. These results illustrate that caution is needed when interpreting the results of questionnaires and measurements of ventilatory lung function in the diagnosis of asthma among working populations. PMID- 8241480 TI - Screening for skin cancer: experience of an occupational health screening programme. AB - A skin cancer screening programme involving 2150 employees based at the head office of a large UK retailer resulted in the detection of four melanomas at an early curable stage. In addition, three other malignant and three potentially malignant tumours were discovered, and individuals at greater risk of developing melanoma were identified and counselled accordingly. PMID- 8241481 TI - Effects of changing the pattern of sickness absence referrals in a local authority. AB - The high costs of sickness absence in North Tyneside Metropolitan Borough Council prompted a review of the system of referral of cases of long-term sickness absence to their occupational health service for opinion about fitness to work. Earlier and more consistent referral of these employees produced reductions in average lengths of sickness absence. Time off before medical retirement was reduced from 72 weeks to 53 weeks, and time off before returning to work was reduced from 40 weeks to 25 weeks. Exact figures of the financial savings could not be calculated, but the estimated saving was 760,000 pounds in the first year. Although the occupational health service was not responsible for the whole of this saving, it played an important role in the exercise. It was concluded that earlier referral of employees with long-term sickness absence enabled decisions about returning to work to be made sooner, thus saving large amounts of money. PMID- 8241482 TI - Planning for medical provision on-board ship. AB - This paper describes the medical provision on board a 76 m passenger vessel. It discusses the floor space necessary, together with essential equipment to provide a high level of medical care particularly in the event of major emergencies. It stresses the necessity for training and practice of simulated emergencies to ensure a high state of readiness. PMID- 8241483 TI - Principles and pitfalls in health services research in occupational health systems. AB - The purpose of this paper is (1) to view the aims and specific objectives of occupational health service systems and (2) to highlight some essential concepts and methods of health systems research in the evaluation and development of occupational health services. None of these tasks will succeed if the objectives of such services are not defined, or if the links with the different interest groups, (employers, employees and state) and with the primary health care are not taken into consideration. PMID- 8241484 TI - Comparison of data on occupational health service systems in Austria, Finland, Germany, The Netherlands, Norway and Sweden. AB - Comparative data on the occupational health systems in different countries are lacking to a very great extent. This is illustrated here with the data from six European countries of participants of the workshop. On the basis of available data and various developments in the countries and the EC, it is concluded that in some countries the coverage of the working population will diminish, while in others it will increase. The need for international comparative research is discussed in the light of the recent EC ruling saying some form of occupational health service should be available for all EC workers. It is concluded that monitoring research is feasible taking the experience in national studies into account. PMID- 8241485 TI - Employees' and occupational physicians' different perceptions of the work relatedness of health problems: a critical point in an effective consultation process. AB - In a survey on workers' attitude and behaviour towards the occupational health service (OHS) in their company, it appears that there is hardly any relationship between physicians' and employees' judgement on the work-relatedness of health problems presented during a consultation hour. This is considered an alarming outcome with severe implication for the doctor-client communication. In this article an explanation for this large discrepancy between the physicians' and employees' judgement is derived from the clarification given by employees and from theories on the lay-definition of health and illness and on doctor-patient interaction. Apart from factors already mentioned in the general literature on the doctor-patient relations, the socio-economic implications of the label of 'work-relatedness of health problems' in the field of the occupational health services, turns out to be an important factor in the explanation. PMID- 8241486 TI - Role of the occupational health services as a part of illness-related primary care in Finland. AB - The aim of the study was to evaluate the employees' illness-related utilization of physicians' services in Finland with special reference to Occupational Health Services (OHS). The data are based on a national health-survey conducted by a Computer Assisted Telephone Interview (CATI) in autumn 1991. There were 1,856 people aged 25-64 interviewed. The "risk" of having visited a physician in six months was in association with perceived chronic illness and gender. Half of those employees who were covered by OHS (90% of all) usually visited their OHS unit for health problems. The potential use of the OHS unit depended on the OHS model. The study shows that the Finnish employees use OHS units for physicians' services a lot and this might have implications in the planned reorganization of the Finnish primary care to the so-called population responsibility-based system. PMID- 8241487 TI - Quality assessment of occupational health services instruments. AB - Interest in the quality of instruments for occupational health services is growing as a result of European legislation on preventive services stressing, for example, risk identification and assessment. The quality of the services can be enhanced when the quality of the applied instruments can be judged and improved. To judge quality aspects of instruments, a basic draft scheme and criteria are presented. A first category of criteria covers measurement objective and study design and comprises for example sample strategy and data aggregation level. Technical quality aspects are validity, reliability, standardization and precision. Acceptability of an instrument by employees, employers and professionals belongs to the process quality. At last, strategic quality or presumed utility has to be assessed. As an illustration four instruments have been judged: periodic occupational health surveys, pre-employment medicals, the company audit on work and health, and specific working conditions assessment instruments. Finally the presented basic scheme and criteria are brought into discussion. PMID- 8241488 TI - Evaluation of an occupational health care programme: negative results, positive results or a failure? AB - Among personnel of the city of Amsterdam the incidence of disability as a result of musculoskeletal problems amounts to 15 per 1000 person-years in the refuse collecting department. A disability prevention programme in accordance with literature was started to decrease this high incidence of disability. In the process-evaluation after 2 years from the start of the project it turned out that only three of the ten preventive measures advocated were realized. These meagre results were explained by strategic factors such as lack of motivation among management and the fact that policy on worker health protection was isolated from the overall company policy. Recommendations for future evaluation research are given. PMID- 8241489 TI - Development and evaluation of an occupational health services programme on the prevention and control of effects of vibration. AB - The outline of a study in occupational health services (OHS) on the prevention and control of effects of whole-body vibration is described and preliminary results are presented here. The study comprises an inventory of current activities of occupational health services in vibration-related issues, the development of a specific OHS-programme and evaluation of this programme, mainly on process level. Results of the inventory, a questionnaire survey in occupational health services (n = 112, response 67%), show that the impact of OHS in prevention and control of adverse health effects is small. The development and evaluation of a specific OHS-programme may be useful in setting up a preventive approach. Some methodologic problems are discussed also. PMID- 8241490 TI - Industrial medicine and the culture of health and safety at work. AB - Prevention in modern day German OHS is not only an additional task. The change from check-up medicine to a holistic design for industrial health and safety has consequences for various levels of everyday routine. Two studies are introduced to describe aspects of the culture of health and safety at work. The study of the Landesinstitut Sozialforschungsstelle analyses the cooperation between Company doctors and Safety-at-Work Inspectors; and the GfAH case study shows some of the practical difficulties that arise when an attempt is made to implement prevention oriented occupational medicine in any one company. PMID- 8241491 TI - Supply and demand factors in occupational health; determinants of self-reported compliance with the work place related requirements of the German Work Safety Law. AB - The study evaluates the extend to which West German company doctors reported that they carried out the workplace related activities as required by the German Work Security Law 'ASiG'. Postal questionnaires were completed by 502 of West German company doctors. About one third of those who responded, reported that they carried out the requirements of the German Work Safety Law with regard to workplace-related prevention. Multivariate analysis was applied to discover the factors that influenced compliance. We conclude that investment in supply-side factors (doctors' qualifications, experiences and recourses) are not enough to bring about compliance. More important are the demand-side regulations which directly address the health and safety culture of the companies. PMID- 8241492 TI - Quality and effectiveness of occupational health services: viewpoint of the European Trade Union Confederation. PMID- 8241493 TI - Quality and effectiveness of occupational health service evaluated by the providers. AB - In a debate on the desired occupational health service in The Netherlands all parties involved seemed to be rather unsatisfied with current practice. Therefore a study was set out on the quality of the service to 51 client organizations as seen by the parties involved. In the first phase the viewpoint of the occupational health teams was studied; the second phase among clients was planned later. As it was not feasible to use proper measures of effectiveness, i.e. of outcome in terms of health, the interview was used as method directed at the evaluations of the teams themselves. It focused on the adequacy of the activities of the teams in view of the health problems and risks in the client organization and on the necessary interaction with the clients in order to reach effective service. Although a self-evaluation-method like this has its limitations, results are relevant for the debate on the quality of OHS in The Netherlands. Many occupational health teams want more capacity for preventive activities and to devote more time to these activities to enhance the quality of their service and be more effective. It is argued that for the same purpose the teams themselves should strive for good relations with their clients. PMID- 8241494 TI - Quality and effectiveness of occupational health services: viewpoint of an international company. PMID- 8241495 TI - Hematopoietic growth factors: understanding functional diversity in structural terms. PMID- 8241496 TI - Effect of all transretinoic acid in newly diagnosed acute promyelocytic leukemia. Results of a multicenter randomized trial. European APL 91 Group. AB - We designed a multicenter randomized trial comparing chemotherapy with daunorubicin-Ara C (chemotherapy group) and all transretinoic acid (ATRA) combined to the same chemotherapy (ATRA group) in newly diagnosed APL patients aged 65 years or less. The major endpoint of the study was event-free survival (EFS) ("events" being defined as failure to achieve complete remission [CR], occurrence of relapse, or death in CR). Early termination of the trial was decided after the first interim analysis, as EFS was significantly higher in the ATRA group. At the time, 101 patients had been randomized (54 in the ATRA group and 47 in the chemotherapy group). In the ATRA group, 49 (91%) patients achieved CR, 5 (9%) had early death, and 0 had resistant leukemia, compared with 38 (81%), 4 (8%), and 5 (10%) patients, respectively, in the chemotherapy group. The difference in CR rate between the two groups was not significant. The duration of coagulopathy was significantly reduced in the ATRA group, compared with the chemotherapy group. In the ATRA group, six patients relapsed after 7 to 15.5 months. In the chemotherapy group, 12 patients relapsed after 1 to 16 months, and 2 died in CR. Kaplan-Meier EFS was estimated at 79% +/- 7% and 50% +/- 9% at 12 months, respectively, in the ATRA and the chemotherapy group (P = .001). Kaplan Meier estimate of relapse was 19% +/- 8% and 40% +/- 12% at 12 months (P = .005). In conclusion, ATRA followed by chemotherapy increases EFS in newly diagnosed APL. These results strongly suggest that ATRA should be incorporated in the front line therapy of newly diagnosed APL. PMID- 8241497 TI - G6PD Nara: a new class 1 glucose-6-phosphate dehydrogenase variant with an eight amino acid deletion. AB - In the course of molecular studies on Japanese glucose-6-phosphate dehydrogenase (G6PD) variants using single-strand conformation polymorphisms (SSCP) analysis, we found an unusual class 1 G6PD variant that had nucleotide deletion in exon 9. The patient showed chronic nonspherocytic hemolytic anemia associated with frequent episodes of severe hemolytic attack. The hemolysate exhibited no measurable activity. Although the partially purified enzyme had detectable activity, we could not perform kinetic studies because of its extreme instability. Nucleotide sequencing showed a unique 24 bp deletion at nucleotide 953-976 that predicts an eight amino acid deletion of TKGYLDDP at residue 319 326. While this is one of the most drastic structural alterations found in G6PD variants, the region with the amino acid deletion was distant from both the G6P and NADP+ binding sites and was located in a domain with low sequence homology among species. The comparatively low functional importance of the deleted region may have saved the patient from lethal tissue dysfunction. PMID- 8241498 TI - Fatal Epstein-Barr virus-associated hemophagocytic syndrome. AB - A virus-associated hemophagocytic syndrome is characterized by high fever, liver dysfunction, coagulation abnormalities, pancytopenia, and a benign histiocytic proliferation with prominent hemophagocytosis in bone marrow, lymph node, spleen, and liver. We describe six Japanese children with fatal Epstein-Barr virus (EBV) associated hemophagocytic syndrome. Five of the six patients had serologic evidence of primary EBV infection at the onset of their diseases. EBV genomes were detected in all the patients by Southern blot hybridization or the polymerase chain reaction. Furthermore, clonality analysis of the EBV genome showed that EBV-infected cells proliferated monoclonally or biclonally in three examined patients. In situ hybridization study using EBV-encoded RNA 1 (EBER1) showed that EBER1 was detected in one of two examined liver tissues, which localized in hepatocytes. PMID- 8241499 TI - Phase I trial with recombinant human interleukin-3 in patients with lymphoma undergoing autologous bone marrow transplantation. AB - Recombinant human interleukin-3 (rhIL-3) was administered to 30 patients undergoing autologous bone marrow transplant (ABMT) for treatment of lymphoma. In this phase I dose escalation study, rhIL-3 was administered from day 0 to 20 after ABMT by 2-hour intravenous infusion at dose levels of 1, 2, 5, and 10 micrograms/kg/d. Seventeen patients did not complete therapy with rhIL-3. Eleven requested early discontinuation for malaise, confusion, transplant complications, or rapid engraftment and were removed from the study, whereas six patients developed grade III toxicity, including fever (three patients), or headache (three patients) possibly attributable to rhIL3. Other common toxicities included diarrhea, rigors, mucositis, and rash. The maximum tolerated dose of rhIL-3 was 2 micrograms/kg/d. No evidence of earlier hematopoietic cell recovery was observed compared with similar historical patients treated with recombinant human granulocyte-macrophage colony-stimulating factor. Future trials will be needed to determine alternate schedules of administration of rhIL-3 or the use of rhIL-3 in combination or in sequence with other growth factors. PMID- 8241500 TI - Identification and characterization of a high-affinity granulocyte-macrophage colony-stimulating factor receptor on primary rat oligodendrocytes. AB - We previously showed the presence of receptors for granulocyte-macrophage colony stimulating factor (GM-CSF) on tumor tissues and tumor cell lines that are derived from the neural crest. To determine whether normal neural cells express functional GM-CSF receptors, we isolated and analyzed primary rat brain cells, including microglia, astrocytes, and oligodendrocytes. Scatchard analysis of equilibrium binding of 125I-GM-CSF to primary rat oligodendrocytes showed an average of 1,110 GM-CSF binding sites per cell, with a kd of 20 pmol/L. In six separate experiments, no specific binding was detectable on the astrocyte population. Microglia were used in competitive binding experiments with oligodendrocytes, and addition of microglia did not increase the specific binding of labeled ligand to oligodendrocytes. In dose-response assays, we measured 3H thymidine uptake in rat oligodendrocytes, microglia and control murine 32D cells stimulated with various concentrations of GM-CSF. Over concentration ranges of 0.025 to 1000 pmol/L, cell proliferation and peak 3H-thymidine incorporation was observed at approximately 30 pmol/L for both the control cells and the oligodendrocytes. However, the microglial cells did not proliferate in response to GM-CSF. These data indicate the presence of a functional receptor for GM-CSF on primary rat oligodendrocytes, and suggest that hematopoietic growth factors such as GM-CSF may play a role in nerve cell development, function, or response to injury. PMID- 8241501 TI - Human granulocyte-macrophage colony-stimulating factor receptor signal transduction requires the proximal cytoplasmic domains of the alpha and beta subunits. AB - Human granulocyte-macrophage colony-stimulating factor (GM-CSF) controls the production, maturation, and function of cells in multiple hematopoietic lineages. These effects are mediated by a cell-surface receptor (GM-R) composed of alpha and beta subunits, each containing 378 and 881 amino acids, respectively. Whereas the alpha subunit exists as several isoforms that bind GM-CSF with low affinity, the beta common subunit (beta c) does not bind GM-CSF itself, but acts as a high affinity converter for GM-CSF, interleukin-3 (IL-3), and IL-5 receptor alpha subunits. The cytoplasmic region of GM-R alpha consists of a membrane-proximal conserved region shared by the alpha 1 and alpha 2 isoforms and a C-terminal variable region that is divergent between alpha 1 and alpha 2. The cytoplasmic region of beta c contains membrane proximal serine and acidic domains. To investigate the amino acid sequences that influence signal transduction by this receptor complex, we constructed a series of cytoplasmic truncation mutants of the alpha 2 and beta subunits. To study these truncations, we stably transfected the IL-3-dependent murine cell line Ba/F3 with wild-type or mutant cDNAs. We found that the wild-type and mutant alpha subunits conferred similar low-affinity binding sites for human GM-CSF to Ba/F3, and the wild-type or mutant beta subunit converted some of these sites to high-affinity; the cytoplasmic domain of beta was unnecessary for this high-affinity conversion. Proliferation assays showed that the membrane-proximal conserved region of GM-R alpha and the serine-acidic domain of beta c are required for both cell proliferation and ligand-dependent phosphorylation of a 93-kD cytoplasmic protein. We suggest that these regions may represent an important signal transduction motif present in several cytokine receptors. PMID- 8241502 TI - Comparison of the human and mouse erythropoietin genes shows extensive homology in the flanking regions. AB - Considerable insights into important cis regulatory elements in a gene can be gleaned from the identification of sequence homologies among different species. To extend and optimize the sequence comparison between human and mouse erythropoietin (Epo) genes, we have obtained new human sequence from 5,547 to 385 bp upstream of the cap site and extended the 3' flank by 489 bp. In addition, we have obtained new sequence information on the mouse Epo gene extending from within the 3' untranslated region (UTR) to 1,001 bp downstream of the polyadenylation site. Analysis of these additional sequences shows considerable homology between human and mouse Epo genes as far as 4 kb (human) or 3 kb (mouse) upstream of the cap sites, as well as far more homology at the 3' end than was previously realized. In addition, both species were found to have a high frequency of short interspersed (SINE) repetitive sequences that interrupt homologies in both the 5' flank and within the transcription unit. PMID- 8241503 TI - Mouse strain variability in the expression of the hematopoietic stem cell antigen Ly-6A/E by bone marrow cells. AB - The cell surface molecule Ly-6A/E provides a convenient marker for primitive stem cells in the hematopoietic tissues of both fetal and adult mice. However, previous studies have shown that Ly-6A/E expression by lymphocytes is variable depending on the haplotype of the Ly-6 locus. Therefore, strain-specific variation in Ly-6A/E expression by bone marrow (BM) cells was investigated. The results show that Ly-6a mice have, on average, 50% of the number of BM cells expressing Ly-6A/E relative to that for Ly-6b mice. Furthermore, among the 5% of BM cells that do not express antigens characteristic of mature T, B, myeloid, or erythroid lineages, which include the primitive hematopoietic stem cell compartment, Ly-6a mice have, on average, more than fivefold fewer Ly-6A/E+ cells relative to that for Ly-6b mice. Isolation of Ly-6A/E- and Ly-6A/E+ cells from mice of both haplotypes showed that, whereas 99% of the marrow repopulating activity (MRA) of C57BL/Ka (Ly-6b) mice could be recovered in the Ly-6A/E+ fraction, only about 25% of the MRA of BALB/c (Ly-6a) was recoverable in the same population. On a per-cell basis, the Ly-6A/E+ cells that were isolated from BALB/c mice were essentially equivalent in MRA to those isolated from C57BL/Ka mice. Thus, whereas a large percentage of the hematopoietic stem cells of Ly-6a mice do not express the Ly-6A/E molecule, the antigen may be used to isolate a subset of stem cells from these mice. These results show that hematopoietic stem cell phenotype can vary between mouse strains and imply that caution should be exercised in the identification of human stem cell antigens such as CD34, because a similar variability may occur between individual humans. To further explore the influence of Ly-6 haplotype on Ly-6A/E expression by specific cell subsets, lymph node lymphocytes from a panel of mouse strains were analyzed by multiparameter flow cytometry for correlated expression of Ly-6A/E, CD4, and CD8. All Ly-6a strains examined had less than 20% Ly-6A/E+ cells, and those cells were predominantly CD8+ T lymphocytes. In contrast, the Ly-6b strains had greater than 30% Ly-6A/E+ cells, and those cells included CD4+, CD8+, and B lymphocytes. PMID- 8241504 TI - The 47-kD fragment of talin is a substrate for protein kinase P. AB - This laboratory has been characterizing protein serine/threonine kinase reactions of hematopoietic tissues, whose most distinguishing characteristics in vitro are stimulation with vesicular phosphatidyl glycerol, and the ability to function using Mn2+ as the sole divalent cation. The major protein substrates are a 73-kD protein and a protein migrating near ovalbumin on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The 47-kD protein was partially purified from cells harvested by leukapheresis from a patient with acute myelogenous leukemia, using ammonium sulfate precipitation and ion exchange chromatography. This partially purified ion-exchange fraction contained an endogenous kinase activity with characteristics similar to those we previously described of protein kinase P (protein kinase, phospholipid-stimulable: PK-P), but not typical of any form of protein kinase C (PK-C). With longer phosphorylation, the 47-kD band showed increasingly lower mobility demonstrable both by Coomassie blue staining and autoradiography, suggesting both that it was multiply phosphorylated, and that the excisable band was pure. The protein was thus eluted from preparative gel slices and digested with endoproteinase lys C. Sequence data from the fragments identified the protein as the 47-kD calpain fragment of talin, a protein found in focal adhesion plaques and some cell-cell contacts. PK-C phosphorylated the 47-kD protein, as has been reported previously, and phosphopeptide mapping disclosed a similar pattern of phosphorylation using either PK-C or the endogenous activity. The 47-kD protein labeled with the endogenous kinase contained predominantly phosphoserine, with some phosphothreonine and a trace of phosphotyrosine. Intact, purified talin was also phosphorylated by PK-P in a phospholipid-stimulable manner, but at 1/20 the rate of the 47-kD fragment. PMID- 8241505 TI - Mechanisms of thrombin generation during surgery and cardiopulmonary bypass. AB - Although in vitro studies have been invaluable in revealing the complex biochemistry of the blood coagulation system, the mechanisms involved during the in vivo response to hypercoagulable stimuli are still unclear. We have used plasma-based enzyme-linked immunosorbent assays (ELISAs) to study the mechanisms by which the coagulation system is activated in vivo during human cardiopulmonary bypass (CPB) surgery (n = 8). A novel immunoassay for factor XIIa was used to detect activation of the contact system, factor IX activation peptide (FIXAP) was used as a marker for activation of factor IX, and prothrombin fragment F1 + 2 (F1 + 2) was used as a marker for thrombin generation. The ELISA for FIXAP is described for the first time herein. F1 + 2 levels increased early in response to surgical intervention: from a baseline of 38.7 +/- 9.7 ng/mL (mean +/-SE), levels increased rapidly during surgery and bypass to a maximum of 448.5 +/- 92.0 ng/mL. A modest yet significant increase in factor XIIa levels from 3.47 +/- 0.54 ng/mL to 4.33 +/- 0.85 ng/mL was evident during surgery before bypass, but no further significant increase was detected on establishing extracorporeal circulation. FIXAP levels demonstrated a small and late increase during surgery from 4.98 +/- 0.55 ng/mL to a maximum of 10.20 +/- 1.23 ng/mL, the increase beginning at the time of near maximal F1 + 2 levels. There was no association between activation of the contact system (factor XIIa levels) and the generation of thrombin (F1 + 2 levels). However, a strong association (r = .705) was apparent between the generation of thrombin (F1 + 2 levels) and activation of factor IX (FIXAP levels), despite the delay between the activation of prothrombin and factor IX. The data do not support the established view that contact activation resulting from exposure of blood to foreign surfaces is the major procoagulant stimulus in CPB. Instead, the results suggest that the main trigger to coagulation during CPB surgery was provided via the tissue factor-factor VIIa mechanism in response to the cutting of blood vessels, which directly activated factor X and then prothrombin. The late activation of factor IX, which presumably also contributed to maximal prothrombin activation, could have arisen due to direct tissue factor factor VIIa action, or by secondary feedback action of thrombin on the intrinsic system. PMID- 8241506 TI - Glycoprotein IIb and IIIa retention on fibrinogen-coated surfaces after lysis of adherent platelets. AB - The platelet-membrane glycoprotein IIb-IIIa (GPIIb-IIIa) complex is essential for platelet aggregation and is involved in the attachment of platelets to thrombogenic surfaces. This study shows the retention of GPIIb and GPIIIa on immobilized fibrinogen after Triton X-100 (Sigma Chemical Co, St Louis, MO) lysis of adherent platelets. Glycoproteins were detected using subunit specific monoclonal antibodies in a modified enzyme-linked immunosorbent assay procedure. GPIIb-IIIa retention was judged to be specific relative to GPIb recovery, and was modulated by platelet activation. Platelet exposure to adenosine diphosphate or thrombin, but not A23187 or chymotrypsin, markedly enhanced GPIIb and GPIIIa recovery relative to that observed with unstimulated platelets, or prostaglandin E1-treated platelets. Moreover, lysis of adherent platelets in the presence of 10 mmol/L EDTA, under conditions promoting GPIIb-IIIa complex dissociation (pH 8.1, 60 minutes, 37 degrees C), had no effect on GPIIb or GPIIIa subunit recovery. Platelet activation with Zn+2 also enhanced GPIIb and GPIIIa recovery on fibrinogen-coated surfaces over that observed with unstimulated platelets, but GPIIb and IIIa retention was EDTA sensitive. This correlated with the EDTA reversible nature of Zn+2-activated platelet adhesion to fibrinogen-coated surfaces. The data (1) show that platelet adhesion to fibrinogen is accompanied by the induction of high-affinity interactions between GPIIb-IIIa and immobilized fibrinogen that are EDTA-resistant and enhanced by platelet activation with some but not all agonists, and (2) implicate these interactions in stabilizing platelet contacts with fibrinogen-coated surfaces. PMID- 8241507 TI - C1-inhibitor-serine proteinase complexes and the biosynthesis of C1-inhibitor by Hep G2 and U 937 cells. AB - The biosynthesis of the serpin alpha 1-proteinase inhibitor is regulated by a feedback mechanism whereby complexes between alpha 1-proteinase inhibitor and serine proteinases bind to liver cells and monocytes, a reaction that activates alpha 1-proteinase-inhibitor gene transcription. Such a mechanism may form the basis for the development of new therapeutic strategies for serpin deficiency states with reduced levels of otherwise normally functioning serpins. This issue was addressed for C1-inhibitor, the missing serpin in hereditary angioedema. C1 inhibitor biosynthesis by Hep G2 hepatoma cells was assessed by enzyme-linked immunosorbant assay, by metabolic labeling followed by immunoprecipitation, and by Northern blotting. C1-inhibitor biosynthesis was stimulated by gamma interferon (100 U/mL) but not by cell exposure to C1-inhibitor-kallikrein (1 mumol/L), C1-inhibitor-C1s (1 mumol/L), and C1-inhibitor-plasmin complexes (1 mumol/L) or to reactive site-cleaved C1-inhibitor (1 mumol/L). Moreover, radioiodinated C1s-C1-inhibitor complex did not bind to Hep G2 cells. C1 inhibitor-kallikrein complex was also without effect on C1-inhibitor mRNA in U 937 cells. Therefore, the proposed mechanism, by which serpin-enzyme complex or reactive site-cleaved serpin binding to a specific receptor provides a signal for the stimulation of the biosynthesis of that serpin, is not operative for the biosynthesis of C1-inhibitor by Hep G2 or U 937 cells. PMID- 8241508 TI - Plasma dermatan sulfate proteoglycan in a patient on chronic hemodialysis. AB - A 68-year-old man on chronic hemodialysis for 6 years, presented with a spontaneous psoas muscle hemorrhage. Investigations showed intermittently elevated activated partial-thromboplastin time and thrombin time. Preliminary investigations suggested a heparin-like inhibitor in the patient's plasma, but no anti-Xa activity could be detected. Investigation of the ability of patient plasma to inhibit exogenous thrombin showed that most thrombin was inhibited by heparin cofactor II, in contrast to normal plasma in which most thrombin was inhibited by antithrombin III. Treatment of plasma with glycosaminoglycan degrading enzymes suggested the presence of dermatan sulfate (DS) in patient plasma. This was confirmed in a heparin cofactor II-dependent antithrombin assay for DS that showed anticoagulant equivalent to 2.2 +/- 0.3 micrograms/mL (mean +/ SD) of porcine mucosal DS. Of this activity, approximately 90% was sensitive to enzymes that degrade DS. The glycosaminoglycan containing fraction of plasma was isolated and subjected to gel chromatography. Anticoagulant activity eluted from Sephadex G-100 (Pharmacia, Montreal, Quebec, Canada) as two peaks with Kav of 0.10 and 0.45. After treatment with base, the Kav of the higher molecular weight species was increased to 0.55. This activity was completely sensitive to enzymes that degrade DS. Thus, the active DS was present as a proteoglycan. The lower molecular weight material was not sensitive to enzymes that degrade DS or heparan sulfate and it was active in the heparin cofactor II-dependent antithrombin assay but not in an antithrombin III-dependent antithrombin assay. This activity was not degraded by heating. Subsequently, measurement of DS activity was performed in plasmas obtained from eight other patients on hemodialysis before administration of heparin that showed that all patients had DS activity present that varied from 0.05 to 0.4 microgram/mL. No enzyme-resistant activity could be shown in these patients. In summary, a circulating anticoagulant with properties of DS is present in patients requiring hemodialysis. PMID- 8241509 TI - Molecular genetics of myeloid leukemia: identification of the commonly deleted segment of chromosome 20. AB - A deletion of the long arm of chromosome 20 [del(20q)] is a recurring abnormality in malignant myeloid disorders. The occurrence of the del(20q) in a broad spectrum of myeloid disorders suggests that the loss of genetic material on 20q could provide a proliferative advantage to myeloid cells, possibly through the loss of a tumor-suppressor gene. We have examined a series of patients with the del(20q) using fluorescence in situ hybridization (FISH) with unique sequence probes that map along the length of 20q, and have delineated a segment that is deleted in 95% of all patients examined (18 of 19). In addition, we have shown that the deletions are interstitial rather than terminal. This region of deletion extends from 20q11.2 to q12, and is flanked by the RPN2 (proximal) and D20S17 loci (distal). The SRC and ADA genes are located within the commonly deleted segment. Our findings emphasize the importance of FISH and other molecular mapping techniques in defining such a region. The delineation of a commonly deleted segment in 20q11.2-q12 will facilitate the identification of candidate tumor-suppressor genes on 20q. PMID- 8241510 TI - Expression and release of CD27 in human B-cell malignancies. AB - CD27, a transmembrane disulfide-linked 55-kD homodimer, belongs to the nerve growth factor-receptor family, a group of homologous molecules involved in lymphocyte differentiation and selection. It is expressed on mature thymocytes, peripheral blood T cells, and a subpopulation of B cells. We investigated the expression of CD27 on malignant B cells representative for a broad range of stages in physiologic antigen-independent and -dependent B-cell development. In normal lymphoid tissue CD27+ B cells were only found in the peripheral blood (29.8% +/- 10.8%, n = 13) and in germinal centers. With the exception of pro-B and the majority of pre-pre-B acute lymphocytic leukemias and of myelomas, CD27 expression of variable intensity was detected on almost all immature and mature malignant B cells tested. Moreover, using a sandwich enzyme-linked immunosorbent assay we could show the presence of sometimes very high (up to 6,000 U/mL; normal values < 190 U/mL) amounts of the soluble 28- to 32-kD form of CD27 (sCD27) in the sera of patients with B-cell malignancies. The highest levels of sCD27 were observed in patients with chronic lymphocytic leukemia and low-grade non Hodgkin's lymphomas. Most importantly, both in transversal and longitudinal studies, we found a strong correlation between sCD27 levels in the serum and tumor load, indicating that sCD27 can be used as a disease-marker in patients with acute and chronic B-cell malignancies. PMID- 8241511 TI - p53 gene mutation in B-cell chronic lymphocytic leukemia is associated with drug resistance and is independent of MDR1/MDR3 gene expression. AB - We studied 53 patients with B-cell chronic lymphocytic leukemia (B-CLL) and found mutations of the p53 gene in 15%. Patients with p53 gene mutations were found to have an aggressive form of B-CLL disease characterized by advanced Rai stage, rapid lymphocyte doubling time (LDT), and resistance to chemotherapy. While 27 of 29 treated patients (93%) without p53 mutations achieved a partial remission, only one of seven treated patients (14%) with p53 mutations achieved a partial remission (P = .00009). Adjusting for prognostic factors (age, sex, race, and Rai stage), patients with p53 gene mutations had a 13-fold greater risk of death than patients without p53 mutations (P = .013). In addition to examining the clinical relevance of p53 gene mutations in B-CLL, we investigated the possible role of p53 gene regulation in the expression of the multidrug resistance genes MDR1 and MDR3. We quantitated MDR1 and MDR3 mRNA expression by reverse transcription polymerase chain reaction (RT-PCR). Expression of both the MDR1 and MDR3 genes was independent of p53 gene mutation or prior drug treatment, and did not predict for clinical response. Our findings indicate that p53 gene mutations in B-CLL are associated with a poor clinical outcome and may be a prognostic indicator for drug resistance. PMID- 8241512 TI - Regulation of human neutrophil aggregation: comparable latent times, activator sensitivities, and exponential decay in aggregability for FMLP, platelet activating factor, and leukotriene B4. AB - We have recently described a flow cytometry technique, whose sensitivity allows direct measurements of latent times before the onset of aggregation, and of rates, maximal extents, and reversibility of aggregation (J Leuk Biol 50:434, 1991). We report here that activators which stimulate sustained cellular signaling associated with increases in intracellular calcium (ionomycin) or protein kinase C activation (phorbol myristate acetate, PMA) cause complete (> or = 98%) and irreversible neutrophil aggregation, with latent times for the onset of aggregation inversely proportional to the activator concentration. In contrast, the receptor-specific activators leukotriene B4 (LTB4), formyl peptide FMLP, and platelet-activating factor (PAF) gave only partial and reversible aggregatory responses, limited by the following similar properties: latent times of 4.5 seconds +/- 1.5 seconds, independent of activator concentration; similar concentrations for onset of aggregation (approximately 1 nmol/L) that increased over a similar broad range of activator concentration, with one-half maximal rates of aggregation at 10 nmol/L to 30 nmol/L, corresponding to reported dissociation constant values; comparable limited recruitment and spontaneous reversibility of aggregation; absence of interactivator synergism; and similar exponential decays in activated cell stickiness (refractoriness), with t1/2 = 15 to 30 seconds. Variable cross-desensitization was seen between LTB4 and FMLP depending on donor and activator concentrations. In vivo, these properties are expected to provide localization of the aggregatory response, minimizing the otherwise detrimental effects of circulating activated neutrophils. PMID- 8241513 TI - Senescence of canine biotinylated erythrocytes: increased autologous immunoglobulin binding occurs on erythrocytes aged in vivo for 104 to 110 days. AB - We have evaluated senescence related changes in canine red blood cells (RBCs) using the biotinylation system, where RBCs are labeled in vivo with biotin at the beginning of their life span, and retrieved from circulation on immobilized avidin at the end of their life span. This approach avoids the controversial use of density gradient centrifugation to collect presumably old RBCs. Furthermore, the dog is an appropriate model for human RBC senescence because it has a low degree of random RBC loss and a similarly long RBC life span (approximately 110 days). Two dogs had 97% to 100% of their circulating RBCs biotinylated by infusion of N-hydroxysuccinimido biotin (Clontech, Palo Alto, CA; Calbiochem, La Jolla, CA) dissolved in dimethyl sulfoxide. At postbiotinylation days 104 and 107 for one dog and day 110 for the other dog, biotinylated RBCs were isolated by magnetic cell sorting and analyzed for the presence of autologous IgG using 125I labeled sheep-antidog IgG (SAD IgG). On all 3 days, there were at least three times more SAD IgG molecules per RBC on senescent biotinylated RBCs than on control (unfractionated) RBCs (day 104: 11,677 v 3,399; day 107: 6,710 v 2,115; day 110: 6,042 v 1,838 molecules of SAD IgG per senescent v control RBC). Furthermore, it is unlikely that an immune response to the conjugated biotin had been elicited, because fresh in vitro biotinylated RBCs that were incubated in autologous plasma (taken after exposure to circulating biotinylated RBCs for 113 days) and then exposed to the SAD IgG showed no increase in antibody binding over control (non-biotinylated) RBCs (1,431 v 1,378 cpm/10(8) biotinylated v control RBCs; P > .20). These results suggest that senescence of canine biotinylated RBCs is characterized by binding of autologous IgG and that antibiotin antibodies do not contribute to this process. PMID- 8241514 TI - Hemoglobin S polymerization and gelation under shear II. The joint concentration and shear dependence of kinetics. AB - The kinetics of hemoglobin S gelation are critical in sickle disease because microvascular obstruction can be avoided if red blood cells pass these vessels during the delay time, before polymerization and gelation occur in sufficient degree to rigidify the cells. Kinetics, including the delay time and the closely related exponential progress rate, are highly sensitive to hemoglobin concentration and degree of deoxygenation. Kinetics are also greatly accelerated by shear, an effect that may contribute to pathogenesis, since red blood cells deform and can undergo shear in vivo. Here we examine the joint dependence of kinetics on shear and hemoglobin concentration. As shear rate increases, the concentration dependence of the exponential progress rate decreases. The large decrease in concentration dependence supports the conclusion that acceleration of gelation by shear is due to breakage and not to enhancement of heterogeneous nucleation. Under shear, new fibers are created by breakage of existing ones, as well as by heterogeneous nucleation. At high shear, the rate of new fiber creation by breakage is very great and dominates that by heterogeneous nucleation. Therefore, if breakage depended only on shear rate and were independent of the concentration of hemoglobin in solution, the concentration dependence of kinetics should vanish. Although it decreases, it does not disappear. The concentration dependence that remains at high shear arises from (1) the direct contribution of fiber growth rate to the exponential progress rate, (2) the dependence of breakage rate on fiber growth rate, and (3) the dependence of solution viscosity on hemoglobin concentration. PMID- 8241515 TI - Membrane attachment sites for the membrane cytoskeletal protein 4.1 of the red blood cell. AB - The identity of the membrane binding sites for the membrane cytoskeletal protein 4.1 of the human red blood cell has been investigated. Exhaustive proteolysis of the membrane with a range of proteases led to the elimination of only some 60% of all binding sites. The predominant integral membrane protein, band 3, as well as glycophorin A, was totally digested at levels of proteolysis that were essentially without effect on the number of 4.1 binding sites. Proteolysis caused scission of the polypeptide chain of glycophorin C (together with the minor product, glycophorin D, of the same gene), but left a fragment from the region of the C-terminus still attached to the membrane. We have found a low-molecular weight protein, possessing an epitope (recognized by an antibody directed against the cytoplasmic domain of glycophorin C) in common with this proteolytic fragment, in cells of a Leach phenotype, which are characterized by lack of extracellular epitopes of glycophorin C. When these membranes were extracted at low ionic strength to dissociate the membrane cytoskeleton, approximately half the content of 4.1 was liberated, compared with only some 25% from normal membranes. Cells of a different variant of the Leach phenotype, which are totally devoid of glycophorin C, lost close to 70% of their 4.1 under these circumstances. The Rh(D) transmembrane protein, which interacts with the membrane cytoskeleton, is also resistant to proteolysis of the cytoplasmic membrane surface, but Rhnull cells, devoid of this protein, showed no decreased retention of 4.1. The results suggest that glycophorin C (with D) may contain two types of binding site for 4.1, which would be sufficient in number to account for all the strong binding of 4.1 on normal membranes; modulation of binding at one of the sites by another protein or by lipid is not excluded. A possible site for reinitiation of translation overlapping the premature stop codon in the mutant expressing the truncated glycophorin C can be discerned. PMID- 8241516 TI - Assessment of the hemostatic effectiveness of human platelets treated with aminomethyltrimethyl psoralen and UV A light using a rabbit ear bleeding time technique. AB - The photochemical aminomethyltrimethyl psoralen (AMT), in conjunction with UV A light (UVA), has been shown to inactivate human immunodeficiency virus-1 and model viruses in platelet suspensions under conditions that have only a minimal effect on in vitro platelet properties. A rabbit ear bleeding time technique was used to assess the hemostatic effectiveness of human platelet suspensions treated with AMT/UVA. New Zealand White rabbits were made thrombocytopenic by a combination of irradiation and heterologous antirabbit platelet antiserum. Reticuloendothelial function in these rabbits was suppressed by the intravenous administration of ethyl palmitate. The hemostatic function of 1- and 5-day-old human platelet suspensions (14.5% plasma) that had been treated on day 1 with 40 micrograms/mL AMT and 24 kJ/m2 UVA (1 x UVA) was evaluated by measuring microvascular bleeding times after a standard incision. Comparable bleeding times were observed after infusion with both control and AMT/UVA-treated platelets stored for either 1 or 5 days. With the transfusion of AMT/1 x UVA-treated platelets stored for 5 days, the mean (+/- SD) bleeding time was 156.3 +/- 39.2 seconds (n = 10). With untreated platelets (no AMT/no UVA), stored for 5 days, the mean bleeding time was 189.2 +/- 36.4 seconds (n = 10). Neither AMT nor 1 x UVA treatment alone influenced the observed bleeding times. In contrast, the hemostatic effectiveness of human platelet suspensions was diminished if they were exposed to three times the standard UVA dose (72 kJ/m2) on day 1 and stored for 4 more days, regardless of whether AMT was present, with the mean bleeding time increasing to 442.2 +/- 122.6 seconds (n = 15, AMT present) or 396.0 +/- 45.9 seconds (n = 10, AMT absent). These results are consistent with data obtained from in vitro studies and indicate that virucidal AMT/1 x UVA treatment does not influence platelet hemostatic function. However, the final conditions to achieve these results must be carefully controlled. PMID- 8241517 TI - Immunoglobulin levels and monoclonal gammopathies in children after bone marrow transplantation. AB - Bone marrow graft recipients suffer profound immunodeficiency during at least 3 months after transplantation. B-cell reconstitution following allogeneic bone marrow transplantation (BMT) in children was studied longitudinally by quantification of Ig (sub)class levels in serum and by investigation of numbers and characteristics of homogeneous Ig components (H-Ig); ie, monoclonal gammopathies (MG). For the latter purpose, a sensitive immunoblotting technique capable of detecting H-Ig of a concentration as low as 0.5 microgram/mL was used. Sera of 40 children grafted for a variety of diseases were investigated and followed up for 5 years. It was found that Ig (sub)classes reached normal levels from 3 months after BMT onward. The sequential increase of the different Ig isotypes was in accordance with that seen in normal ontogeny. This was especially clear following BMT for severe congenital immunodeficiency. H-Ig appeared from as early as 6 weeks after BMT in increasing numbers, beginning within IgM, IgG3, and IgG1, and afterward within other isotypes. After an initial increase of serum Ig levels, "overshooting" occurred accompanied by high frequency of H-Ig. H-Ig were still present at 5 years after BMT, when Ig levels normalized. Our data indicate that B-cell reconstitution after allogeneic BMT recapitulates normal ontogeny but in a clonally dysregulated fashion; that is, with overexpression of some clones and underexpression of others. PMID- 8241518 TI - Rapid detection of chain termination mutations in the alpha 2 globin gene. PMID- 8241519 TI - Splenectomy for immune thrombocytopenia. PMID- 8241520 TI - Plasma cell labeling index, beta 2-microglobulin, and C-reactive protein: what is the best combination for myeloma prognosis? PMID- 8241521 TI - Use of an inbred mouse model system for studies of allogeneic transfusion-induced immunosuppression. PMID- 8241522 TI - Cytokine priming of acute myeloid leukemia may produce a pulmonary syndrome when associated with a rapid increase in peripheral blood myeloblasts. PMID- 8241523 TI - Pathophysiology and pharmacology of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is the classical example of an immune complex (IC) associated systemic autoimmune disease. Although an important part of SLE etiopathogenesis has yet to be discovered, it is generally accepted that genetic factors, sex hormones, alternations in T- and B-lymphocyte activity and defects in RES-function contribute to the development of SLE. In an SLE patient, symptoms and severity of the disease are linked to the pattern of autoantibodies expressed, referring to some pathophysiological importance of antibodies found in SLE. In addition, the interindividually variable expression of antibodies to ds DNA, Ro or anticardiolipin, for example, permit a subtyping of SLE and indicate SLE as collective concept of heterogeneous systemic connective tissue diseases with overlapping, e.g. to dermatomyositis, progressive systemic sclerosis or Sjogren's syndrome. In view of the variable, heterogeneous disease manifestations, it is obvious that the strategy in SLE therapy is to treat manifestations and not just SLE per se. Using this concept together with pathophysiologically related control parameters and standard clinical investigations, 90 to 95% of SLE patients are adequately treated with NSAID, steroids, antimalarials and immunosuppressiva. Only 5-10% need experimental therapy, and this kind of treatment should be strictly limited to this group. PMID- 8241524 TI - Bis azo dyes--studies on the mechanism of complex formation with IgG modulated by heating or antigen binding. AB - The mechanism of binding of azo dyes (bis azo) to immunoglobulin G of altered conformation, induced by heating or interaction with antigen was analysed in this work. Azo dyes: Congo Red, Evans Blue and Trypan Blue were selected for these studies. The molecules of Congo Red and Evans Blue associate readily in water and exist as polymolecular micellar species of liquid crystalline organization. Such organization of molecules appeared necessary for these dyes to interact with antibodies and to affect the formation of immune complex. It was proved by studying the properties of isomeric dyes Evans Blue and Trypan Blue, whose ability to form polymolecular conglomerates in water differs, being high for Evans Blue and low for Trypan Blue. The dyes seem to influence the formation of the immune complex generally by interacting with individual immunoglobulin molecules. The rate of exchange of dye molecules in conglomerate bound to protein varies and is the lowest for the portion of molecules which are engaged directly in the complex with protein. Electron microscopic studies also confirmed the polymolecular form of the dye in the dye-protein complex. The computational simulation of dye-dye and dye-protein interaction was performed in a model system. The micelle in the system was represented by three molecules of Congo Red. The peptide loop composed of amino acids 68-88 originating from VL IgG domain, was selected to represent protein. Amino acid side chains of this fragment were reduced to C beta. The best fitness was found for peptide chains of twisted beta conformation and independently optimized conformation of the dye in a form of "twisted ribbon" micelle. It was concluded that the IgG domains become accessible for penetration of the dye after being relaxed in the result of heating or interaction with the antigen. PMID- 8241525 TI - Bis-azo dyes interference with effector activation of antibodies. AB - Azo dyes (Congo Red, Evans Blue, Trypan Blue) were used to study the relation between antigen binding and effector activity in SRBC rabbit anti-SRBC IgG system. Antigen binding was tested in this study by agglutination and effector activity by complement fixation. Antibodies induced by the antigen binding appeared to become structurally susceptible for interaction with the dye. The dyes cause the enhancement of antigen binding by antibodies and inhibition of their effector activity. The intensity of both effects caused by binding the dye seems correlated. The increase of antibody concentration makes the effector activity of immune complexes more resistant to dye inhibition indicating that the influence of antigen binding may be exerted on assembling of antibody in the immune complex rather than on structural modulation of Fc. Also the analysis of dye effects on antibody derivatives deprived of effector activity (Fab)2, IgG with split interheavy chain disulphides supplied evidence that the induced by the antigen long distance structural effects may play the role of immune signal by influencing the assembling of antibodies and stability of immune complex. PMID- 8241526 TI - A comparison between the automatized impedance cardiography and pulsed-wave Doppler echocardiography methods for measurements of stroke volume (SV) and systolic time intervals (STI). AB - Stroke volume (SV) and systolic time intervals (STI) were measured automatically using impedance cardiography signals (ICG) and compared with those obtained by pulsed-wave Doppler echocardiography using the apex approach. The comparison was made in 9 healthy male subjects, mean age 24.9 +/- 12.2 years, using recordings of 10 heart cycles simultaneously obtained by the two methods. During measurements the subjects rested in the supine position. There were no differences between mean values of SV determined by the two methods as well as between mean values of ejection time (ET) (p > 0.8 and p > 0.9, respectively). The pre-ejection period (PEP) estimated by ICG was 22 ms longer than that determined by echocardiography (p < 0.001). The relationship between SV values measured by impedance cardiography (SVA) vs those calculated by echocardiography (SVE) was found to be close to the line of identity in the range of measurements. The regression equation for SV was: SVA = 0.784.SVE + 15 (r = 0.69, p < 0.001, SEE = 10.7 ml). We conclude that automatic determination of SV and ET from ICG signals provides results comparable in absolute values with those obtained by the pulsed wave Doppler ultrasonocardiography using the apex approach for subjects remaining in the supine position. PMID- 8241527 TI - Double-blind crossover study of ranitidine and ebrotidine in gastro-esophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) is multifactorial disorder in which acid exposure has a central role in the mucosal damage, and the mainstay of medical treatment is the suppression of gastric acid secretion justifying the use of H2 receptors antagonists. In our study we compared the effects of ranitidine and ebrotidine, a novel H2 antagonist with gastroprotective properties, on the motor, pH and endoscopic aspects of GERD in randomized cross-over trial in humans. Twenty patients with endoscopic evidence of erosive esophagitis were included in the study. Esophageal manometry and 24-hour pH-metry were done with the use Synectics (Sweden) systems. The same examinations were repeated after 20 days period of treatment with either ranitidine or ebrotidine, given in single dose 300 and 800 mg (nocte) respectively. The pressure within the lower esophageal sphincter (LES) in the untreated and treated with ebrotidine or ranitidine patients remained lowered. Patients with GERD showed increase in duration and decrease in amplitude and propagation of peristaltic waves in the esophageal body which were not improved after treatment. Complete healing after 40 days of treatment was comparable with ebrotidine and ranitidine and averaged about 40%. The pH-metry showed improvement in treated patients in the reflux frequency and time pH below 4, ranitidine being more effective than ebrotidine. It can be concluded that GERD patients showed weaker primary peristalsis unrelated to LES pressure and treatment. Treatment with ebrotidine or ranitidine reduced significantly the endoscopic and self-assessment score, ebrotidine and ranitidine being equally effective in healing of esophageal mucosal lesions. PMID- 8241528 TI - The influence of calcium antagonists (verapamil, nifedipine, and MgCl2) on rat gastric damage induced by ethanol in vivo and in vitro. AB - Mucosal gastric injury was induced in vivo by oral administration of 75% ethanol to rats that were intraperitoneally pretreated with saline (controls), 4 mg/kg of verapamil, 0.4 mg/kg of nifedipine or 160 mg/kg of MgCl2. The glandular stomachs were used for macroscopic and histologic evaluation of mucosal lesions. The gastroprotective effects of substances tested against ethanol-induced mucosal damage in vitro were examined: 1). in rats intraperitoneally pretreated with calcium antagonists and 2). after adding of 10(-4) M verapamil, 10(-6) M nifedipine or 10(-2) M MgCl2 directly to minced gastric mucosa of untreated rats. This effect was measured by DNA synthesis. Gross and histologic evaluation showed that rats pretreated with nifedipine or MgCl2 had significantly decreased ethanol induced gastric injury compared to controls, whereas those pretreated with verapamil had significantly increased injury. On the contrary, all verapamil, nifedipine and MgCl2 treatments, administrated intraperitoneally or by exposure to an incubation mixture were equally effective in reducing gastric mucosal damage following to ethanol treatment in vitro. We conclude that the differential effects of verapamil and nifedipine on ethanol-damaged gastric mucosa in vivo, but not in vitro, suggests the existence of different actions of these calcium antagonists more on systemic rather than local protective mechanisms, such as gastric mucosal blood flow, mucosal barrier, or cell renewal. PMID- 8241529 TI - Changes in the membrane potential can affect pepsinogen secretion of isolated rat chief cells. AB - The aim of this study was to determine pepsinogen secretion by isolated rat chief cells in relation to changes of membrane potential obtained by the use of potassium ionophore valinomycin and different extracellular K+ concentrations, or protonophore carbonyl cyanide m-chlorophenyl hydrazone (CCCP) at different pH. While valinomycin or CCCP decreased basal and cAMP-stimulated secretory response of the chief cells, they increased secretion of pepsinogen stimulated by calcium ionophore A23187. When Ca2+ was absent from the incubation media, A23187 stimulated secretion of pepsinogen significantly decreased. Moreover, the omission extracellular calcium reversed the potentiating effects of valinomycin or CCCP on A23187-stimulated secretory response. It is known that pepsinogen secretion is a biphasic process. In the first phase, Ca2+ is released from its intracellular stores, while the second one depends on extracellular calcium entering the cell. Our results suggest that this phase is dependent also on membrane potential. PMID- 8241530 TI - Thyrotropin-releasing hormone (TRH) modulates vasopressin and oxytocin release from the hypothalamo-neurohypophysial system in dehydrated rats. AB - Rats euhydrated and dehydrated during two or four days were given intracerebroventricularly (i.c.v.) thyrotropin-releasing hormone (TRH) in a daily dose of 200 ng dissolved in 10 microliters of 0.9% sodium chloride. A single dose of TRH injected to euhydrated animals increased the hypothalamic vasopressin content but did not affect significantly the content of vasopressin in the neurohypophysis as well as that of oxytocin both in the hypothalamus and neurohypophysis. In rats deprived of water for two days TRH completely prevented the decrease of neurohypophysial oxytocin due to stimulation of osmoreceptor origin. Similarly, TRH restrained both the hypothalamic and the neurohypophysial vasopressin and oxytocin depletion in rats dehydrated for four days. PMID- 8241531 TI - Central histaminergic mechanisms in the corticosterone response to clonidine. AB - Involvement of central histaminergic mechanisms in stimulation of the hypothalamic-pituitary-adrenal (HPA) axis by clonidine was investigated in conscious rats. Clonidine as well as adrenergic and histamine receptor antagonists were administered intracerebroventricularly (icv), the antagonists always 15 min prior to clonidine, and 1 h later the trunk blood was collected for corticosterone determination. alpha-Fluoromethylhistidine (alpha-FMH), a neuronal histamine synthesis inhibitor, was given ip 2 h before clonidine. Immediately after decapitation, brains were exposed and hypothalami were isolated on ice and frozen for further spectrofluorimetric histamine determination. The clonidine induced increase in the serum corticosterone level was considerably, but not totally, reduced by icv or ip pretreatment with yohimbine, an alpha 2-adrenergic receptor antagonist. The rise in the corticosterone level induced by clonidine was significantly diminished by mepyramine, a histamine H1-receptor antagonist, and moderately lowered by cimetidine, a histamine H2-receptor antagonist. Clonidine significantly augmented the histamine content in the hypothalamus and rest of the brain. The clonidine-induced increase in hypothalamic histamine might be the cause of an increased corticosterone secretion via stimulation of central H1-histamine receptors. On the other hand, alpha-FMH injected 2 h before clonidine considerably diminished both the histamine content in the hypothalamus and the corticosterone secretion induced by clonidine. These results indicate that clonidine given centrally stimulates the HPA activity via not only alpha adrenergic but also histaminergic mechanisms. Clonidine augments the hypothalamic histamine which, in turn, stimulates the corticosterone secretion, predominantly via histamine H1-receptors. Neuronal histamine is considerably involved in the stimulatory action of clonidine since inhibition of the neuronal histamine synthesis by alpha-FMH significantly depresses the corticosterone response to clonidine. PMID- 8241532 TI - [How should we manage chemical substances in living environments]. AB - Recent major advances in science and technology have Introduced a large number of synthetic chemical substances to our societies, resulting in the occasional unfortunate serious pollution of our environment by these chemical substances. A study of the administrative management of these chemical substances has shown inadequacies, as has been revealed in the case of countermeasures against carcinogenic substances in the environment which have not been managed systematically. In view of this situation, the rapid introduction of the risk assessment-risk management process in Japan is proposed, as an administrative countermeasure against carcinogenic substances. Problems that need to be examined for accelerating the introduction of this process are: scientific development of risk assessment methodology, and a methodological arrangement of risk management process. As for the latter, the level of acceptable risk which determines "virtual safe dose" should be defined, and the role of cost-benefit analysis in risk management should be understood. PMID- 8241533 TI - [Relationship of nutrient intake to serum enzyme activity]. AB - Measurement of the serum enzyme activity is generally used as a screening test for hepatic and biliary diseases. The purpose of this study is to examine the use of serum enzyme activity as indices of health by analyzing relationships to changes in nutrient intakes. The results obtained were as follows: In cross sectional studies gamma-GTP (1) and LDH-4 and -5 isozyme levels had a significant negative correlation to dietary intakes of vitamin C and protein, but total serum gamma-GTP and LDH activities had no correlation. Intake trials of these nutrients showed that high intake levels of vitamin C related to decrease in isozymes levels while low protein intake resulted in increases. Serum GPT activity had a negative relationship to vitamin C intake in men, but was not correlated with nutrient intakes in women. Serum ALP activity was affected positively with vitamin B intake in 40-79 year old men and women. There were statistically significant correlations between serum cholinesterase activity and energy, and food intake patterns. PMID- 8241534 TI - [Subjective well-being differences in disabled women as related to length of bedridden period]. AB - A cross-sectional study of disabled women living at home, aged 65 years and more, was conducted in Osaka-city to examine difference in subjective well-being in two groups differing in length of bedridden period, and to elucidate correlates of subjective well-being. A short-period group was composed of 43 disabled women whose bedridden periods were shorter than two years, and a long-period group of 63 women was randomly selected from disabled women of the corresponding age having a bedridden experience of five or more years. The two groups were interviewed by public health nurses using the Philadelphia Geriatric Center Morale Scale. The subjective well-being was lower in the short-period group than in the long-period group. This result indicates the necessity to provide adequate support to the disabled elderly early on, not only from the physical stand point but also for consideration of subjective well-being. Other correlates of subjective well-being in the disabled women were age, number of family members and caregivers, economic state, pain, ophthalmopathy and utilization of domiciliary care services. Existence of a plan for home visits by public health nurses was strongly related to subjective well-being, possibly indicating that the visiting plan was made with some consideration of subjective well-being. These need to be further development of adequate support systems that consider the subjective well-being of the disabled elderly. PMID- 8241535 TI - [A comparative study of the health conditions of elderly people living alone, elderly couples and the bedridden elderly at home in a rural area of Shiga Prefecture: special reference to morbidity rate and blood pressure, electrocardiograph and blood examination data]. AB - For the purpose of obtaining basic data and for establishing a support system for elderly people with various health and social problems, a population survey was performed in 1990 to investigate the health and living conditions of elderly people living alone, elderly couples and the bedridden elderly in the town of Shigaraki, Shiga Prefecture. A total of 275 subjects (103 male, 172 female) 65 years of age and over were surveyed. The participation rate in this survey was 88.1%. Analysis of health conditions (morbidity rate, blood pressure, electrocardiograph and blood examination data) of elderly people living alone, elderly couples and the bedridden elderly produced the following: 1) Stroke was the main cause of being bedridden in men, while in women, bone and joint disease, especially fracture, was the main cause. 2) The combined prevalence of hypertension and borderline hypertension in elderly people living alone, elderly couples and the bedridden elderly was over 50%. The bedridden elderly had a lower prevalence than elderly people living alone and elderly couples. 3) Men in all of the above mentioned life styles, had a higher tendency of showing ECG abnormalities than women. The tendency for major ECG abnormalities was high for bedridden elderly, both male and female, with the tendency for men being higher. 4) In bedridden elderly, a tendency of higher prevalence of anemia, in both male and females, lower total serum cholesterol and triglyceride in males compared to elderly people living alone and in elderly couples, was observed. PMID- 8241536 TI - [A longitudinal study on health status and factors relating to it in elderly residents of a community]. AB - The purpose of the present study is to clarify the relationship of physical, sociological and psychological factors to health status, the decline in activities of daily living (ADL) and death, in community elderly residents, by analysis of participation or non-participation in a comprehensive health survey. Subjects were 737 elderly residents (306 males, 431 females) aged 70-75 years living at home in Fujishimamachi, a rural town of Yamagata prefecture. A baseline comprehensive health survey including door-to-door survey for non-respondents to the comprehensive survey, was performed in 1986, and repeated at follow-up five years later. The results obtained are as follows: 1) In both sexes, survival rate was highest in the respondents to the comprehensive health survey and lowest in non-respondents to either surveys. Causes of death, place of death, the bedridden period prior to death and ADL before bedridden status were different among those three groups. 2) Factors predisposing to ADL decline (all subjects had competent normal ADL at the baseline survey) were low self-rated health, low score of the Index of Competence developed by Tokyo Metropolitan Institute of Gerontology (the TMIG Index of Competence), inactivity in daily life especially in household affairs and social activities. The predicting factors of death were almost similar to those of ADL decline mentioned above. The relationship of the factors to health status was stronger in those who had to be surveyed door-to-door compared to respondents to the comprehensive health survey, and in women than in men. 3) Blood pressure, ECG findings, retinal findings, hand grip, memory as measured by the Benton Visual Retention Test showed no relationship to either ADL decline or mortality. 4) The present study shows that subjective health and functional capacity are useful to predict the outcome of community elderly residents. Further, life styles and health behavior have a significant effect on outcome prediction. PMID- 8241537 TI - [Alcohol-related problems in mortality in middle-aged urban-dwelling men from cerebro-cardiovascular diseases]. AB - A survey was performed of the life-styles and medical histories of men, who died of cerebro-cardiovascular diseases at ages between 40 and 64 years in Hirakata, Neyagawa, Moriguchi and Kadoma cities in Osaka prefecture. Data was available on 127 of 172 men who died in 1988. Of these, 28 had histories of alcohol-related problems. Analysis showed that the amount of alcohol consumed was associated with disease history and serious alcohol-related problems. 1) Histories of hypertension, cerebrovascular and liver disease were found more often in heavy drinkers with a daily alcohol intake exceeding the equivalent of 46 g pure ethanol compared to either moderate drinkers with a daily alcohol intake of less than 46 g or abstainers. 2) Of 28 subjects with alcohol-related problems, one had been treated at a specialized alcoholic-clinic, two had been admitted to psychiatric hospitals and one subject had consulted a physician at a mental health clinic of the public health center in his community. 3) Although 18 of the 28 subjects with alcohol-related problems had their own regular doctors, they did not appear to have received adequate care. 4) Subjects with alcohol-related problems had histories of cerebrovascular and liver diseases more often than those without alcohol-related problems. 5) Subjects with alcohol-related problems were confronted with unemployment, divorce and housing problems more often than those without alcohol-related problems. With the increasing amount of alcohol consumption in Japan, alcohol-related problems need more attention. There is an urgent need to establish community-based strategies for prevention of alcohol related problems and to organize a network of multi-disciplinary support teams for those with problems such as alcoholics in urban communities. PMID- 8241538 TI - [Studies on the reduction and correction risk factors for arteriosclerosis in junior high school pupils]. AB - Circulatory system examinations performed on first-grade pupils of junior high schools (12 years old; N = 1,204), identified 105 high-risk pupils with arteriosclerosis-promoting factors. These pupils were consulted on life-style modification, emphasizing nutrition and exercise, and encouraged to practice personal health management and self-monitoring. Reduction and correction of high risk factors were studied in these subjects, based on review of results of health examinations performed regularly up to the time of their graduation. Results are as follows: 1. Averages for obesity grade (for both sexes) in the second and the third years were significantly lower than that in the first year. Based on a general reference value > or = 20%, improvements were observed in 22.5% of males and 50.0% of females. 2. In both sexes average total cholesterol (TC) and triglyceride (TG) were lower in the second and the third years than in the first year, and a significant increase in HDL cholesterol was seen. With regard to a general reference value > or = 3.0 for AI (Atherogenic Index) calculated using TC and HDL cholesterol, improvements (reduction) were observed in 29.4%, of males and 46.2% of females. 3. While the numbers of subjects with elevated SBP, DBP, GPT, FPG and BUN were few, there were significant decreases in averages in the second and third years compared with those in the first year. Using general reference values, improvements in these risks were seen in 100% for all values except for GPT where 75.0% showed improvement. 4. Comparison of grouped distribution of summed values of examination results between the first year and the third year, showed improvements (reduction and correction of risks) in 69 pupils (65.7%), while 30 pupils (28.6%) remained in the same group with no improvement and 6 pupils (5.7%) turned for the worse, dropping one rank.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241539 TI - [Visual acuity testing of three-year-old children using Landolt ring and picture targets]. AB - Visual acuity testing using Landolt ring and picture targets (silhouettes of butterfly, fish, bird and tulip), was performed on 184 three-year-old subjects who had received secondary ophthalmic examination at the age of 3 years and 1-3 months, and the results were evaluated. The examinations were feasible on 83.1% of the subjects for the picture test and 76.1% for the Landolt ring test; the picture test being significantly higher. However, when the level was set at 0.5, namely, 0.5 or more were judged as "passing" and less than 0.5 judged as "failing", the Landolt ring test showed a significantly higher rate than the picture test in the number "passing". Specificity and sensitivity were calculated with reference to the clinical findings. Results suggest that the Landolt ring test has a better validity than the picture test and the Landolt ring test could differentiate between good and poor vision. The silhouette of the bird was particularly poor. When using picture targets for testing, the necessity of proper selection of the kind of pictures used is indicated. In the case of use of the Landolt ring, reexamination at the age of 3 years and 6 months or later, or postponement of the examination until age 3 years and 6 months or later would appear to be beneficial for enhancing the feasibility of the test. PMID- 8241540 TI - [Serum cholesterol level and food intake in old persons in Tottori prefecture]. PMID- 8241541 TI - Electrophysiological analysis of transmission at the skeletal neuromuscular junction. AB - The review is divided into two sections. The first deals with methods and problems associated with performing electrophysiological experimentation on the skeletal muscle neuromuscular junction. The second section concentrates on the computer analysis of electrophysiological data. In the first section, the various techniques available for producing skeletal muscle paralysis are described. These include the use of pharmacological manipulations, such as an excess of magnesium ions or a competitive postjunctional nicotinic acetylcholine antagonist, physiological manipulations, such as cutting the muscle fibers, and the muscle fiber sodium channel toxin, mu-conotoxin. Also, in this section, a comparison is made of the use of voltage- and current-recording techniques, including descriptions of, and solutions to, the problems associated with membrane capacitance, nonlinear summation, membrane space constant, and electrical and mechanical interference. In the second section, details are given of the types of computer system commonly used for the analysis of electrophysiological data and also the requirements of the data analysis software. The use of computer algorithms for signal detection, signal evaluation, signal averaging, and curve fitting are qualitatively described, along with some of the problems and pitfalls often associated with these methods. PMID- 8241542 TI - A novel in vivo model for the study of cartilage degradation. AB - Methods of quantifying cartilage destruction are described using a sponge/cartilage implant model in the rat. A cylinder of bovine nasal cartilage was positioned in the center of a sponge which had been pretreated with an irritant. The sponge/cartilages were then implanted subcutaneously into the backs of rats for periods of up to 16 days. The implanted sponges were rapidly surrounded by granulation tissue, maximal on day 2, and infiltrated by inflammatory cells which reached peak levels on day 9. Analysis of the cartilage shows an initial increase in wet weight and rapid loss of glycosaminoglycans. These changes were later followed by loss of cartilage wet weight and significant loss of hydroxyproline content. In a separate study, the effects of Mycobacterium tuberculosis (Mtb), kaolin, and zymosan were compared (1 mg/sponge) and the results showed that only Mtb induced pronounced inflammation and degradation of cartilage. The cartilage degradation directly correlated with the granulation tissue weight, but not with cellular infiltration. We believe that this simple, reproducible in vivo model could be used to elucidate the mechanisms involved in the destructive process and evaluate the efficacy of inhibitors of cartilage degradation. PMID- 8241543 TI - A method for estimation of extracellular concentration of compounds by microdialysis using urea as an endogenous recovery marker in vitro validation. AB - A new, practical method called microdialysis has been developed for the estimation of free, unbound drug concentrations in the interstitial fluid. This method is based on the use of urea as an endogenous recovery marker. Urea freely distributes throughout the body water compartment, and its plasma concentration can be used as an accurate measure of its interstitial levels. Microdialysis exploits the relationship between the relative recoveries of urea and the recoveries of an analyte that are established during calibration. The calibration is carried out using different probes, varying perfusion rates, membrane surface areas, temperatures, and tissue models. After obtaining a broad range of recoveries, an empirical mathematical model of the relationship between the recovery of the respective analyte and that of urea is formed. During in vivo experiments, a microdialysis probe is placed into a tissue or fluid of interest, and the urea recovery is monitored by comparing the corresponding dialysate and plasma urea levels. From the calculated urea recoveries, correlation equation, and analyte dialysate concentrations, the extracellular concentration of the unbound analyte in the tissue or fluid can be estimated at any given time point. The purpose of this study is to describe the method of microdialysis and demonstrate its applicability in vitro using two model compounds, theophylline and 3'-azido-3'-deoxythymidine (AZT), in the rat plasma. PMID- 8241544 TI - Simultaneous assessment of cardiac output with pulsed Doppler and electromagnetic flowmeters during cardiac stimulation. AB - The aim of this study was to test the accuracy of cardiac output assessment by Doppler and electromagnetic flowmetry in dogs during states of (1) marked enhancement in cardiac output, which was obtained by means of either isoprenaline infusion or treadmill exercise, or (2) reduction in cardiac output obtained by administration of phenylephrine. Additionally, in vitro comparisons were undertaken between Doppler and electromagnetic flow-probes and assessment of flow by direct volumetric measurement. These in vitro experiments showed a good correlation between timed volume collections and electromagnetic flow assessment up to high flow velocities. Doppler flow measurements underestimated the flow at high velocities. In both the resting dog and after phenylephrine, that is, at states with low heart rate and cardiac output, the waveforms of electromagnetic flow and Doppler velocity were similar for both phasic and mean flow, respectively. During states of cardiac stimulation Doppler flow showed a decrease in maximum velocity in the ascending aorta. Due to this decrease in peak flow velocity, mean Doppler blood flow did not increase despite of increased heart rate. This result cannot be explained on the basis of the deviation of Doppler measurements at high velocities in the in vitro experiments. Although our results are in contradiction with earlier studies, electromagnetic assessment seems to be more reliable in blood flow measurements in the ascending aorta. Hence, Doppler flow measurements should not be used uncritically for such quantitative flow assessment in large vessels as determination of cardiac output. PMID- 8241545 TI - Regional blood flow as a determinant of drug absorption description of an animal model. AB - Factors that influence recommended dosage guidelines for intramuscularly and subcutaneously injected drugs include a spectrum of variables, including therapeutic objective(s), the drug's therapeutic index, the inclusion of concomitant vasoconstrictors, and so on. However, little account is taken of the likely influence of regional blood flow at the injection site. This may vary according to the site of injection, the age and medical condition of the patient with the potential to markedly affect the rate of drug absorption and the resultant therapeutic and toxicity outcomes. An animal model is described for simultaneously quantifying the drug's pharmacokinetic profile and the regional blood flow (using technetium99 elution) at sites of differing vascularity so that the potential relationship can be ascertained. An application to lignocaine absorption is provided, however, the model would be applicable to a range of drug applications and could serve as a means of investigating differences both within a drug class (e.g., members with differing lipophilicity) as well as across drug classes (e.g., local anesthetics, opiates, insulin preparations, etc.). PMID- 8241546 TI - Species variation in the effect of glycoprotein IIb/IIIa antagonists on inhibition of platelet aggregation. AB - Differences exist between platelets of different species in their reaction to pharmaceutical agents, such as inhibitors of platelet aggregation. Understanding these differences is critical in the interpretation of data from experimental animal models of thrombosis. Platelet aggregation, essential in the hemostatic process, requires that fibrinogen (fgn) bind to activated platelets. Analogs of Arginine-Glycine-Aspartic acid-Phenylalanine (RGDF), a peptide sequence of fgn, block fgn binding to its receptor known as glycoprotein (GP) IIb/IIIa on activated platelets and prevent aggregation. We studied the inhibition resulting from Arginine-Glycine-Aspartic acid-Serine (RGDS) and two analogs of RGDF, (SC 46749 and SC-47643) on aggregation of human, rat, guinea pig, dog, and rhesus monkey platelets in vitro using ADP as the agonist. The inhibitory potency of RGDS, SC-46749, ad SC-47643 was species dependent. The rank order of potency was rhesus monkey, dogs, and human followed by guinea pig and rat. In order to study the relative inactivity of the compounds in rat platelets compared to human, we diluted rat platelet-rich plasma (PRP) to yield platelet levels approximating that of humans. Platelet inhibition was not significantly changed in diluted rat PRP nor did changing concentration appear to affect activity in human PRP. Our data suggest that the platelet response of some species may better represent human response with regard to inhibition of GP IIb/IIIa by (RGDX) analogs. PMID- 8241547 TI - Aspects of vertebrate gustatory phylogeny: morphology and turnover of chick taste bud cells. AB - The taste bud is a receptor form observed across vertebrates. The present report compares chick taste buds to those of other vertebrates using light and electron microscopy. Unlike mammals, but common to many modern avians, the dorsal surface of chick anterior tongue lacks taste papillae and taste buds. Ultrastructurally, chick buds located in the anterior floor of the mouth (as in some reptiles and amphibians) and palate contain dark, intermediate, light, and basal cell types. Dark, intermediate, and light cells extend microvilli into intragemmal lumina and pores communicating with the oral cavity. As specialized features, dark cell apices lack dense granules and exhibit short microvilli relative to light and intermediate cells. Dark cell cytoplasmic fingers envelop intragemmal nerve fibers and cells as in other species, and sometimes contain abundant clear vesicles. Nerve profile expansions often are located adjacent to dark, intermediate, and light cell nuclei. Classical afferent synaptic contacts are rarely observed. Taste cell turnover is suggested by mitotic and degenerating figures in chick buds. In addition, tritiated thymidine injected into hatchlings, whose anterior mandibular oral taste bud population approximates that in adults, reveals a turnover rate of about 4.5 days. This is about half that observed in altricial mammals, reflecting a species difference or developmental factor in precocial avians. It is concluded that chick taste buds exhibit morphologic features common to other vertebrate buds with specializations reflecting the influences of niche, glandular relations, and/or age. PMID- 8241548 TI - Pre- and postnatal development of rabbit foliate papillae with special reference to foliate gutter formation and taste bud and serous gland differentiation. AB - The epithelial downgrowth arises in the presumptive foliate papilla region of the tongue approximately at prenatal day 22, and the distal portion of the primary epithelial cell cords transforms to the bifurcated serous gland and excretory duct between prenatal day 30 and postnatal day 2. The excretory ducts extend upward through the primary epithelial cell cords at rather random intervals and are open to the tongue surface between postnatal days 1 and 2. In this process, successive connections between the adjacent ascending excretory ducts occur mainly due to desquamation of the keratinized lining cells of the ducts, resulting in the formation of the foliate gutter. Taste bud primordia appear in the primary epithelial cell cords by nerve penetration through the basal lamina from prenatal day 30 and nearly fully formed taste buds facing the ascending excretory ducts have been already observed before the foliate gutter formation is completed. In the differentiation process of chemoreceptor (type III) cells from less-differentiated basal (type IV) cells, subsurface cisterns of endoplasmic reticulum are occasionally present where nerve endings make contact. Since subsurface cisterns have been considered to be involved in reciprocal or efferent synaptic transmission, it is reasonable to consider that these morphological specializations may take a role in neurotrophic signals for differentiation of the chemoreceptor cells. PMID- 8241549 TI - Ultrastructural evidence for a binding substance to the sweet-tasting protein thaumatin inside taste bud pores of rhesus monkey foliate papillae. AB - Thaumatin is a protein that tastes intensely sweet only to Old World monkeys and to higher primates, including man. Here we used pre-embedding ultrastructural methods to study the distribution of thaumatin in apical regions of Rhesus monkey foliate papillae, using thaumatin conjugated to 5 nm gold particles. With freeze substitution we saw that gold-labeled thaumatin bound to an electron-opaque, sponge-like secretory substance inside the taste bud pores. Labeled thaumatin was found at the surface of the secretory substance even deep inside the pore, where other, unlabeled cellular structures surrounded the substance. With freeze fracture deep-etching the secretory substance that bound the thaumatin-gold particles appeared coarsely granular. There was no labeling of any other taste bud pore structure, including microvilli and small membrane-lined vesicles. Pre incubation with an excess of unlabeled thaumatin inhibited binding with gold labeled thaumatin. The results suggest that the secretory substance had the greatest affinity of all taste pore structures to the sweet-tasting compound under our experimental conditions. Therefore, gustatory reception probably involves various taste compound binding structures, microvilli, and also secretory substances like the one described here which bound thaumatin. We speculate that the secretory substance may bind taste stimuli and serve as an intermediate between stimuli and receptors. It could be involved in stimulus removal or delivery or both. PMID- 8241550 TI - HVEM ultrastructural analysis of mouse fungiform taste buds, cell types, and associated synapses. AB - We have used high voltage electron microscopy and computer-generated three dimensional reconstructions from serial sections to elucidate the structure of taste bud cells and their associated synapses in fungiform taste buds of the mouse. Five fungiform taste buds (two of which were serially sectioned) were examined with the high-voltage electron microscope (HVEM). We identified the synaptic connections from taste cells onto sensory nerve fibers and classified the presynaptic taste cells based on previously established ultrastructural criteria. From those data we have distinguished dark, intermediate, and light cells in murine fungiform taste buds. Synapses in murine fungiform taste buds are fewer in number, but contain many more vesicles than synapses in either foliate or circumvallate taste buds. Synapses in mouse circumvallate and foliate taste buds typically contain a few to several synaptic vesicles per section, whereas fungiform synapses may have in excess of 100 vesicles per profile. The significance of these differences in the numbers of synapses and synaptic structure between fungiform and circumvallate/foliate synapses is not known. Based on the small number of synapses observed in fungiform taste buds, we speculate that fungiform taste buds have only a few cells transducing sensory stimuli at any given time. Alternatively, communication of sensory information from the taste receptor cells to the afferent nerve fibers may be mediated by some other mechanism(s) in addition to classical chemical synapses. PMID- 8241551 TI - Sample preparation technique for cross-sectional transmission electron microscopy of quantum wire structures. AB - A novel cross-sectional sample preparation technique for quantum wire (QWR) structures is described. By coating a thin layer of Au with a designed pattern on the sample as a marker to indicate the position of the wire pattern, the location of the thinned area can be controlled precisely. An example of applying this technique to an InGaAs/GaAs QWR structure is demonstrated. This technique can also be applied to any other small dimensional structures or devices with specific regions of interest. PMID- 8241552 TI - Selected area polishing for precision TEM sample preparation. AB - A selected area mechanical polishing technique has been developed to improve the precision of cross-sectional TEM sample preparation, based upon the early work of Benedict and colleagues [Benedict et al. (1990) MRS Symp. Proc. Vol. 199, p. 189]. TEM samples were made from a pre-selected section through the middle of a 1 micron wide band of transistors extending laterally for more than 1 mm by precise control over the plane of polish with a corresponding reduction in sample preparation time. To illustrate the application of this technique, a uniformly thin, electron transparent TEM sample of a single, specific, failed transistor is obtained from a 4 mm by 10 mm device array. PMID- 8241553 TI - A modified autowasher device for rapidly washing large numbers of EM-grids. AB - This device consists of a siphon system and 5 or 10 grid disks, modified from the previous mode (Chen, 1973), for large numbers of grids with ultrathin sections. This method improves the ease of assembling grids onto the grid disk and also requires much less stain solution. This system only takes 5 min for one single stain washing, at a maximum of 100 grids, and also avoids stain contamination. The grid disk can also be used for immunocytochemistry work and for critical point drying of grids with biological specimens. PMID- 8241554 TI - Use of dental wax for the study of insect behavior by scanning electron microscopy. PMID- 8241555 TI - A simple specimen holder for EBIC imaging on the Hitachi S800. PMID- 8241556 TI - Cell culture supports for slam-frozen and molecular distillation dried procedures. PMID- 8241557 TI - Evidence for stimulus access to taste cells and nerves during development: an electron microscopic study. AB - We have examined developing taste buds in fungiform papillae of rats from the 18th day of gestation (E18) to the 15th postnatal day (P15). Nerve processes were seen in the epithelium of E18 rats before taste buds were obvious. At E20, early taste buds were visible, but were embedded within the epithelium, i.e., their cells were shielded from the oral cavity by overlying squamous epithelium. At this stage, the epithelium on the lateral aspects of the fungiform papillae was keratinized, but that overlying the taste bud was not. Some taste bud cells at E20 contained synapse-like structures near their contacts with nerve processes. In postnatal animals, keratinized epithelial cells were seen overlying taste buds, but taste pores were not observed until P10. How, then, do stimuli reach the taste cells and elicit physiological and behavioral responses as reported by others? The keratinized epithelium overlying the buds was unlike that on the lateral aspect of the papilla in at least one significant way. Few lamellated bodies were present in intercellular spaces beneath the stratum corneum, whereas these were abundant in the corresponding location within epithelium on the slope of the papilla. Although some were present within the squamous epithelium overlying the bud, they apparently were not released into the intercellular space. These lipid-rich lamellated bodies are thought to represent the water barrier of the epithelium, i.e., the barrier which prevents aqueous solutions from passing through the epithelium. We determined that the keratinized epithelium overlying the taste bud was permeable to a tracer, lanthanum nitrate, whereas that on the lateral surface was not. Lanthanum was visualized around taste cells and around nerve profiles within and near the taste bud. We propose that the absence of an aqueous permeability barrier in the epithelium overlying taste buds likely explains the ability of tastants to reach the taste bud cells and nerves in the developmental period before pore formation. PMID- 8241558 TI - Morphometric and immunocytochemical assessment of fungiform taste buds after interruption of the chorda-lingual nerve. AB - Unilateral interruption of the chorda-lingual nerve led to a loss of most epithelial axons and to the deterioration of fungiform taste buds in the anterior portion of the tongue of albino rats, mongolian gerbils, and golden hamsters. By three weeks after surgery the following percentages of fungiform taste buds had completely disappeared: 71% in gerbils, 28% in rats, and 26% in hamsters. Residual taste buds were classified into two groups: atrophic taste buds and taste bud remnants. Atrophic taste buds were smaller than normal and typically had no visible taste pore, although they retained the characteristic oval shape of a taste bud and numerous elongated cells. Taste bud remnants were non-oval fragments of taste buds with few elongated cells. Specific markers for elongated taste cells (monoclonal antibodies to keratin 19) confirmed that atrophic taste buds, as well as some taste bud remnants, had elongated taste cells. By 180 days after chorda-lingual nerve transection, 44% of rat fungiform taste buds had disappeared; morphometric analysis of the 311 residual taste buds established that 241 atrophic taste buds and 69 taste bud remnants were, respectively, 50% and 75% smaller than the average volume of 480 normal taste buds. The aggregate loss of gustatory tissue, calculated from the shrinkage of residual taste buds and the volume lost by the outright disappearance of many taste buds, was 88% for gerbils, 72% for rats, and 65% for hamsters. Evaluation in gerbils of the co occurrence of taste buds and axons suggests residual taste buds were neurotrophically supported. Every gerbil fungiform papilla that lacked axons lacked a taste bud.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241559 TI - Transcellular and paracellular pathways in lingual epithelia and their influence in taste transduction. AB - The lingual epithelium is innervated by special sensory (taste) and general sensory (trigeminal) nerves that transmit information about chemical stimuli introduced into the mouth to the higher brain centers. Understanding the cellular mechanisms involved in eliciting responses from these nerves requires a detailed understanding of the contributions of both the paracellular and transcellular pathways. In this paper we focus on the contribution of these 2 pathways to the responses of salts containing sodium and various organic anions in the presence and absence of amiloride. Electrophysiological recordings from trigeminal nerves, chorda tympani nerves, and isolated lingual epithelia were combined with morphological studies investigating the location (and permeability) of tight junctions, the localization of amiloride-inhibitable channels, and Na-K-ATPase in taste and epithelial cells. Based on these measurements, we conclude that diffusion across tight junctions can modulate chorda tympani and trigeminal responses to sodium-containing salts and rationalize the enhancement of taste responses to saccharides by NaCl. PMID- 8241560 TI - Cellular relations in mouse circumvallate taste buds. AB - The fine structure of the taste buds of circumvallate papillae of two strains of mice was studied by electron microscopy. Mice anesthetized with ketamine were perfused through the heart with a double aldehyde mixture in cacodylate buffer and the tissues embedded in Epon. Semi-serial sections were employed. The morphology and relationships of cell types are consistent with the majority of descriptions of mammalian taste buds served by the ninth cranial nerve. Cells of type II are particularly well documented, as the stages in their origin, maturation and degeneration could be followed. Significant differences, however, relate to cell type I. These cells contain large dense-cored granules, contrasted with the more irregular and somewhat larger dark granules of the type I cells in the rabbit. These granules do not produce a dense homogenous product for the pore, as seen in the rabbit. Rather the pore substance consists of small, empty vesicles in a diffuse dark matrix. These granules are only moderately larger than the dense-cored vesicles of the type III cells. All features of the type III cell were demonstrated, although no instance of a complete cell was seen in any section. No significant differences were noted between the two strains of mice. Intimate proximity of a nerve to a cell nucleolus, suggestive of a trophic pathway, is illustrated. PMID- 8241561 TI - Organization of the nucleus of the solitary tract in the hamster: acetylcholinesterase, NADH dehydrogenase, and cytochrome oxidase histochemistry. AB - The distribution of acetylcholinesterase (AChE), NADH dehydrogenase (NADHd), and cytochrome oxidase (CO) was determined in the nucleus of the solitary tract (NST) in the golden hamster. Histochemical staining was compared to cytoarchitectonic subdivisions of the NST (Whitehead: J. Comp. Neurol. 276:547-572, 1988) and to terminal fields of primary afferents of the nerves that innervate the tongue. These three histochemical methods resulted in differential staining patterns within the NST that were related to certain subdivisions. Transganglionic transport of horseradish peroxidase (HRP) was used to determine the central projections of the chorda tympani (CT), the lingual branch of the trigeminal (L V), and the lingual-tonsilar branch of the glossopharyngeal nerves (L-IX). Alternate or the same brain sections were processed to reveal transported HRP, and NADHd or AChE levels. Increased staining of the neuropil with NADHd and AChE was coincident with the dense part of the afferent terminal fields of all three nerves in the NST and the laterally adjacent dorsomedial part of the spinal trigeminal nucleus. CO showed this pattern only for the most rostral part of the CT field. The densest AChE staining coincided with gustatory afferent terminal fields. The histochemical staining facilitated the interpretation of the organization of the NST. For example, at caudal levels of the gustatory NST, it is suggested that taste processing is localized predominantly in the medial part of the rostral central, and somatosensory processing in the rostral lateral subdivision. AChE or NADHd staining should facilitate studies of connections, topography, and neuroplastic changes of the gustatory NST. PMID- 8241562 TI - Morphological types of neurons located at taste-responsive sites in the solitary nucleus of the hamster. AB - HRP histochemistry and microelectrode mapping were combined to study the sizes, shapes, and orientations of neuronal cell bodies and dendrites located at sites of taste-elicited single unit activity in the nucleus of the solitary tract (NST). Cells responding to sapid stimulation of the anterior tongue were extracellularly recorded using micropipettes containing HRP. Iontophoretic injection of the marker at the recording sites resulted in small (50-200 microns diameter) opaque zones bordered by a small number (2-15) of neurons with Golgi like filling of their cell bodies, dendrites, and to some extent, their axons. The cell bodies were near (50-250 microns) the injection sites into which they sent labelled dendrites. Two broad categories of neurons were typically filled. Elongate cells had oval- to spindle-shaped cell bodies oriented mediolaterally. Two primary dendrites extended 100-300 microns from the cell body, one medially and one laterally, and branched within a cylindrical dendritic field oriented mediolaterally. A minority of the HRP-filled elongate cells had unusually long rostrally or caudally directed dendritic branches. Stellate cells had oval, round, triangular, or polygonal cell bodies and 3-5 primary dendrites coursing 200-300 microns in all directions and branching as unoriented, spheroidal fields. A minority of stellate cells had relatively unbranched wavy dendrites, resembling tentacles, while others had unusually small cell bodies (10-15 microns diameter), small dendrites, and locally arborizing axons. Of 151 labelled cells, all but 12 were remarkably confined to the rostral NST. Nearly 90% were concentrated in the rostral central cytoarchitectonic subdivision, where stellate cells predominated, or in the rostral lateral subdivision, where elongate cells predominated. These morphological types of neurons, filled at neurophysiological recording sites, are compared with cell types identified in previous light and electron microscopic studies of the cytoarchitecture, connections, and synaptic organization of the gustatory NST. PMID- 8241563 TI - Advantages of digitonin extraction to reveal the intracellular structure of rat glomerular podocytes for high-resolution scanning electron microscopy. AB - Kidneys of anesthetized rats were perfused with digitonin to extract cytosolic proteins of glomerular podocytes so that the remaining intracellular structures could be examined by three-dimensional stereo high-resolution scanning electron microscopy (HRSEM). Cytoskeleton, consisting of microtubules and intermediate filaments, was preserved with each applied concentration of digitonin. High concentrations of digitonin (1.0 mg/ml) produced a corrugated appearance in plasma membranes likely due to the formation of digitonin-cholesterol complexes. At 1.0 mg/ml digitonin, the Golgi complex became vesicularized, and mitochondria were well extracted and their ultrastructure preserved. Lower concentrations of digitonin (0.1 and 0.2 mg/ml) were less disruptive to both the plasma membrane and the Golgi complex. Mitochondria, rough endoplasmic reticulum, coated vesicles, nuclear membrane, and chromatin were well preserved. Extraction with digitonin, at the optimal concentration and perfusion time, simultaneously maintains both the cytoskeleton and membranous organelles inside the cell and provides a method to elucidate the interactions between these two components. Furthermore, digitonin extraction should preserve antigenic sites, thereby allowing the localization of intracellular proteins by backscattered electron imaging of immunogold labels in the scanning electron microscope. PMID- 8241564 TI - A conserved signal transduction pathway regulating the activity of the rel-like proteins dorsal and NF-kappa B. PMID- 8241565 TI - Prolactin inhibits epidermal growth factor (EGF)-stimulated signaling events in mouse mammary epithelial cells by altering EGF receptor function. AB - We have previously shown that lactogenic hormones stimulate epidermal growth factor (EGF) mRNA accumulation in mouse mammary glands in vivo and in mouse mammary epithelial cells (NMuMG line). However, our in vitro studies indicate that the lactogenic hormone prolactin (PRL) completely inhibits EGF-stimulated DNA synthesis. PRL does not alter cholera toxin or insulin-like growth factor-1 stimulated cell growth, thus the inhibition appears to be specific for EGF. Our current studies are designed to evaluate the effects of PRL on EGF-stimulated signaling events in the NMuMG cell line. Cells treated with PRL for 30 min demonstrated a loss of high affinity EGF-binding ability. After long-term PRL treatment (18 h) there was a decrease in EGF receptor (R) number, as determined by [125I]EGF binding. PRL treatment (8 h) also decreased EGF-R mRNA levels. An EGF-stimulated increase in EGF-R mRNA observed 2-4 h after treatment was decreased when PRL was added to the cultures. Furthermore, levels of EGF stimulated tyrosine phosphorylation of the EGF-R (170 kDa) and phospholipase C gamma (145 kDa) are dramatically decreased in cells treated with PRL. Also of great interest was a decrease in EGF-stimulated c-myc mRNA in PRL-treated cells. We conclude that PRL is acting to down-regulate the EGF-R, thus limiting EGF stimulated cell signaling in mammary tissue. PMID- 8241566 TI - Mos induces the in vitro activation of mitogen-activated protein kinases in lysates of frog oocytes and mammalian somatic cells. AB - Mitogen-activated protein kinases (MAPKs) are rapidly and transiently activated when both quiescent Go-arrested cells and G2-arrested oocytes are stimulated to reenter the cell cycle. We previously developed a cell-free system from lysates of quiescent Xenopus oocytes that responds to oncogenic H-ras protein by activating a MAPK, p42MAPK. Here, we show that the oncogenic protein kinase mos is also a potent activator of p42MAPK in these lysates. Mos also induces p42MAPK activation in lysates of activated eggs taken at a time when neither mos nor p42MAPK is normally active, showing that the mos-responsive MAPK activation pathway persists beyond the stage where mos normally functions. Similarly, lysates of somatic cells (rabbit reticulocytes) also retain a mos-inducible MAPK activation pathway. The mos-induced activation of MAPKs in all three lysates leads to phosphorylation of the pp90rsk proteins, downstream targets of the MAPK signaling pathway in vivo. The in vitro activation of MAPKs by mos in cell-free systems derived from oocytes and somatic cells suggests that mos contributes to oncogenic transformation by inappropriately inducing the activation of MAPKs. PMID- 8241567 TI - Molecular and genetic analyses of the Caenorhabditis elegans dpy-2 and dpy-10 collagen genes: a variety of molecular alterations affect organismal morphology. AB - We have identified and cloned the Caenorhabditis elegans dpy-2 and dpy-10 genes and determined that they encode collagens. Genetic data suggested that these genes are important in morphogenesis and possibly other developmental events. These data include the morphologic phenotypes exhibited by mutants, unusual genetic interactions with the sqt-1 collagen gene, and suppression of mutations in the glp-1 and mup-1 genes. The proximity of the dpy-2 and dpy-10 genes (3.5 kilobase) and the structural similarity of their encoded proteins (41% amino acid identity) indicate that dpy-2 and dpy-10 are the result of a gene duplication event. The genes do not, however, appear to be functionally redundant, because a dpy-10 null mutant is not rescued by the dpy-2 gene. In addition, full complementation between dpy-2 and dpy-10 can be demonstrated with all recessive alleles tested in trans. Sequence analysis of several mutant alleles of each gene was performed to determine the nature of the molecular defects that can cause the morphologic phenotypes. Glycine substitutions within the Gly-X-Y portion of the collagens can result in dumpy (Dpy), dumpy, left roller (DLRol), or temperature sensitive DLRol phenotypes. dpy-10(cn64), a dominant temperature-sensitive DLRol allele, creates an Arg-to-Cys substitution in the amino non-Gly-X-Y portion of the protein. Three dpy-10 alleles contain Tc1 insertions in the coding region of the gene. dpy-10(cg36) (DRLol) creates a nonsense codon near the end of the Gly-X Y region. The nature of this mutation, combined with genetic data, indicates that DLRol is the null phenotype of dpy-10. The Dpy phenotype results from reduced function of the dpy-10 collagen gene. Our results indicate that a variety of molecular defects in these collagens can result in severe morphologic changes in C. elegans. PMID- 8241568 TI - Gradients in the concentration and assembly of myosin II in living fibroblasts during locomotion and fiber transport. AB - Assembly and motor activity of myosin II affect shape, contractility, and locomotion of nonmuscle cells. We used fluorescent analogues and imaging techniques to elucidate the state of assembly and three-dimensional distribution of myosin II in living Swiss 3T3 fibroblasts. An analogue of myosin II that was covalently cross-linked in the 10S conformation and unable to assemble served as an indicator of the cytoplasmic volume accessible to 10S myosin II. Ratio-imaging of an analogue that can undergo 10S-->6S conversion versus the volume indicator revealed localized concentration of assembly-competent myosin II. In stationary serum-deprived cells and in cells locomoting at the edge of a wound, it was most concentrated in the peripheral cytoplasm, where fibers containing myosin II assemble, and least concentrated in the perinuclear cytoplasm, where they disassemble. Furthermore, fluorescence photobleaching recovery showed myosin II to be less mobile in the periphery than in perinuclear cytoplasm. These results indicate a gradient in the assembly of myosin II. Three-dimensional microscopy of living cells revealed that fibers containing myosin II were localized in the cortical cytoplasm, whereas myosin II was diffusely distributed in the deeper cytoplasm, suggesting that myosin II is assembled preferentially near the cell surface. Localized protein phosphorylation may play a role, because a kinase inhibitor, staurosporine, abolished the gradient of myosin II assembly. PMID- 8241570 TI - Translational suppression of syndecan-1 expression in Ha-ras transformed mouse mammary epithelial cells. AB - A cell surface proteoglycan, syndecan-1, has been shown to participate in the maintenance of the epithelial cell morphology. A point mutated activated c-Ha-ras gene under the control of the glucocorticoid inducible MMTV-LTR promoter was transfected into the mouse mammary epithelial cell line, NOG-8. The NOG-8 ras cells were used to study changes in syndecan-1 expression during epithelial transformation. NOG-8 ras cells, when induced to express Ha-ras, transformed and formed foci in monolayer cultures and colonies in suspension cultures. Expression of syndecan-1 at the cell surface was markedly reduced in cells showing the transformed phenotype. The accumulation of newly synthesized core protein of syndecan-1 was suppressed in these cells, whereas mRNA levels remained unchanged. This novel finding indicates that syndecan-1 expression is translationally suppressed in the Ha-ras-transformed epithelial cells. Hence, syndecan-1 loss during epithelial transformation could take place without altering syndecan gene transcription and, on the other hand, could be one of the critical events involved in malignant transformation. PMID- 8241569 TI - Epidermal growth factor stimulates the disruption of gap junctional communication and connexin43 phosphorylation independent of 12-0-tetradecanoylphorbol 13 acetate-sensitive protein kinase C: the possible involvement of mitogen-activated protein kinase. AB - We previously reported that epidermal growth factor (EGF) induced the disruption of gap junctional communication (gjc) and serine phosphorylation of connexin43 (Cx43) in T51B rat liver epithelial cells. However, the cascade of events linking EGF receptor activation to these particular responses have not been fully characterized. Furthermore, the serine kinase(s) acting directly on Cx43 remain unidentified. In the current study, we demonstrate that downmodulation of 12-0 tetradecanoylphorbol 13-acetate (TPA)-sensitive protein kinase C (PKC) activity does not affect EGF's ability to reduce junctional permeability or phosphorylate Cx43 in T51B cells. EGF in the presence or absence of chronic TPA treatment stimulated marked increases in Cx43 phosphorylation on numerous sites as determined by two-dimensional tryptic phosphopeptide mapping. Computer-assisted sequence analysis of Cx43 identified several protein kinase phosphorylation consensus sites including two sites for mitogen-activated protein (MAP) kinase. EGF stimulated activation of MAP kinase in a time- and dose-dependent manner where the kinetics of kinase activity corroborated its possible involvement in mediating EGF's effects. Moreover, purified MAP kinase directly phosphorylated Cx43 on serine residues in vitro. Two-dimensional tryptic and chymotryptic phosphopeptide mapping demonstrated that the in vitro phosphopeptides represented a specific subset of the in vivo phosphopeptides produced in response to EGF after chronic TPA treatment. Therefore, EGF-induced disruption of gjc and phosphorylation of Cx43 may be mediated in part by MAP kinase in vivo. PMID- 8241571 TI - Centromere function on minichromosomes isolated from budding yeast. AB - Centromeres are a complex of centromere DNA (CEN DNA) and specific factors that help mediate microtubule-dependent movement of chromosomes during mitosis. Minichromosomes can be isolated from budding yeast in a way that their centromeres retain the ability to bind microtubules in vitro. Here, we use the binding of these minichromosomes to microtubules to gain insight into the properties of centromeres assembled in vivo. Our results suggest that neither chromosomal DNA topology nor proximity of telomeres influence the cell's ability to assemble centromeres with microtubule-binding activity. The microtubule binding activity of the minichromosome's centromere is stable in the presence of competitor CEN DNA, suggesting that the complex between the minichromosome CEN DNA and proteins directly bound to it is very stable. The efficiency of centromere binding to microtubules is dependent upon the concentration of microtubule polymer and is inhibited by ATP. These properties are similar to those exhibited by mechanochemical motors. The binding of minichromosomes to microtubules can be inactivated by the presence of 0.2 M NaCl and then reactivated by restoring NaCl to 0.1 M. In 0.2 M NaCl, some centromere factor(s) bind to microtubules, whereas other(s) apparently remain bound to the minichromosome's CEN DNA. Therefore, the yeast centromere appears to consist of two domains: the first consists of a stable core containing CEN DNA and CEN DNA binding proteins; the second contains a microtubule-binding component(s). The molecular functions of this second domain are discussed. PMID- 8241572 TI - Origin and development of the International Federation of Associations of Anatomists. AB - The foundation and the growth of the International Federation of Associations of Anatomists are reported and photographs of the past-presidents are included. The past-president report and the future of Anatomy are outlined. PMID- 8241573 TI - [Radiculo-medullary arteries of vertebral origin. Various anatomic aspects from 30 injections-dissections]. AB - Among medullary arteries we have only examined those directly arising from the vertebral artery at the cervical level. This study is about thirty anatomical samples of cervical spine removed during necropsies; there are treated by injection-dissection method. The observed variations do not allow isolation of any artery for the cervical bulge, as possible most frequently at the lumbar bulge level. We observed variations as sometimes alternate pedicles right or left, as sometimes a symmetrical aspect finishing at the same medullar level. By injecting differently colored liquids, in the right and in the left, a side prevalence has sometimes been noted but real conclusions cannot be draw without isobar and simultaneous injections. PMID- 8241574 TI - [Development of the auditory ossicles in the human embryo: correlations with data obtained in mice]. AB - The comparison of human middle ear ossicles developmental features with morphological and experimental data obtained in the mouse embryo reveals some points of convergence. The stapes develops mainly form Reichert's cartilage, but the malleus and incus derived partially from the first arch cartilage (Meckel's) and from a Reichert's cartilage-derived blastema parallel to the tubo-tympanic recess. These results permit to validate the mouse model in teratological studies. PMID- 8241575 TI - [The deep peripatellan arterial circle]. PMID- 8241576 TI - [Morphometric study of the lumbosacral plexus innervating the muscles of the rat pelvic floor]. AB - Motoneurons of the pelvic muscles in the lumbo-sacral segment of the spinal cord have been extensively studied in different species. However, few informations occur in the literature about the individual morphology of each group of neurons and their connections. Thus, in the Rat we have studied the neuronal mapping of 5 pelvic muscles, bulbospongiosus, ischio-cavernosus, levator ani, sphincter ani, sphincter urethrae, using a retrograde tracer, the horseradish peroxidase (H.R.P.). Most of the muscles injected by H.R.P. receive their innervation from the motoneurons located in the homolateral side of the spinal cord. The motoneurons of sphincter ani (+/- 40) and those of the bulbo-spongiosus (+/- 50) have the same morphology. Their dendrites contact contralateral motoneurons. They are situated in the dorso-medial nucleus of the L5-L6 segments of the spinal cord. The motoneurons of the ischio-cavernosus (+/- 80) and those of sphincter urethrae (+/- 40) are situated in the dorsolateral nucleus of the L5-S1 segments of the spinal cord where they form two distinct groups. On the contrary, in the ventro-medial nucleus, the neurons are mixed. The dendrites in the dorso-lateral nucleus form an important longitudinal network. The motoneurons of the levator ani (+/- 50) are situated in the ventral nucleus of the L5-S1 segments. Our works demonstrate the significance of the dendritic networks: they could play a role in the synchronization and the activity of the pelvic muscles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241577 TI - [Morphogenetic role of the trigeminal nerve. A new approach to facial architecture]. AB - Trigeminal nerve acts a main part in morphogenesis and facial pattern. Its contribution to skeletal building and fibrous distribution according to those of different anatomo-functional matrixes allow a radiographic approach of the mesofacial morphology. Excluding anterior exofacial structures, the first statistical results lays the foundations of an accurate and promising architectural cephalometric analysis of mesofacial part. PMID- 8241578 TI - [Cell proliferation in tumor pathology]. AB - There are many ways to analyse and quantify the proliferating activity of a tumor. The different technics are more and more specific of each phase of the cell cycle. This paper firstly describes the basis of each of these technics (mitosis count, DNA staining with various compounds fluorescent or not, incorporation of specific molecules during the phase of DNA synthesis, detection of proliferation antigens by immunological methods). Secondly, the interest of the proliferating fraction knowledge of a tumor is discussed, for various pathologies and according to the technic used. PMID- 8241579 TI - The riddle of the right hemisphere's contribution to the recovery of language. AB - Although it is generally acknowledged that the right hemisphere plays some role in the recovery of language in aphasic patients, the exact nature of this role is still controversial. According to the model of right hemisphere language proposed by Zaidel, the right hemisphere should selectively contribute to the recovery of language comprehension (and in particular of lexical comprehension) but not to the recovery of language production. According to the model of right hemisphere language proposed by Gazzaniga, on the contrary, no language function is selectively supported by the right hemisphere, but the contribution of this hemisphere to the recovery of language varies widely from one individual to another. The aim of this paper is to try and clarify this problem, starting from the basic clinical observations which have given a preliminary shape to the whole issue, and passing then to a survey of results obtained following more recent and specific lines of research. The following conclusions are reached: (1) although the right hemisphere plays (at least in some patients) a definite role in the recovery from aphasia, a greater role is usually played by the undamaged areas of the left hemisphere; (2) the right hemisphere's contribution to the recovery of language seems to concern more the receptive than the expressive components of speech; and (3) the extent of the right hemisphere's contribution varies widely from one individual to another, probably due to individual differences in the hemispheric representation of language. PMID- 8241580 TI - Specific language impairment in children: research findings and their therapeutic implications. AB - This paper reports the findings from a research project investigating a subgroup of specifically language-impaired (SLI) children. The subgroup of SLI children consists of those characterised by persisting grammatical deficits in comprehension and expression of language. The paper summarises the findings in order to highlight the therapeutic implications from the investigations. The main focus of the investigations was to characterise the SLI children's grammatical knowledge of sentence comprehension, specifically their ability to learn the semantic and syntactic properties of verbs. In addition, an investigation of verbal short-term memory (STM) was carried out, and an analysis was undertaken of the expressive morpho-grammatical characteristics of the children. The investigations revealed that the SLI children did not differ in their STM abilities from children carefully matched on language abilities. Thus, the data do not provide support for therapy directed at increasing auditory memory with an aim of improving expression or comprehension of sentences. It is hypothesised that the SLI children have a deficit in syntactic representations and are unable to specify the structural relationships between constituents in syntax. The implications of the study are that this subgroup of SLI children may be unable to use syntactic cues to help learn semantic properties of verbs, but semantic cues may facilitate learning the syntactic properties of verbs. PMID- 8241581 TI - How do communication aid centres provide services to their clients? AB - A survey of the functioning of the six communication aid centres (CACs) in England and Wales was undertaken. All CACs are specialist units drawing clients from a wide area of the country which means that referrals are more likely to come from outside, rather than from within the districts in which the CACs are located. All CACs have developed individual assessment procedures, with some CACs undertaking continuous assessment of clients, and others recommending an aid after a single session. Funding of aids was often a problem because CACs were only able to recommend aid provision and did not supply them; their stocks of aids were used in assessment sessions or for interim loan to clients until a definitive aid had been acquired. CAC staff engaged in a variety of other functions including the organisation of courses, acting as an advice and information service, and undertaking research. PMID- 8241582 TI - The speech of adults with acquired profound hearing losses. I: A perceptual evaluation. AB - Two audio recordings of ten adventitiously profoundly deaf Australian adults and ten age- and sex-matched controls reading a standard passage were played to a panel of ten normally hearing Swedish listeners. One recording consisted of the full speech signal. The second recording presented only the output from a Laryngograph. The listeners were asked to listen to each speaker and then judge whether he or she was adventitiously deafened or normally hearing. When listening to the full speech signal the listeners were able to identify correctly the speakers' hearing status on 92.5% of presentations. The identification rate fell to 65.6% when the listeners only heard the Laryngograph output. If a rating of adventitious deafness was made when listening to the full speech signal, the listeners were asked to state which aspects of the speaker's speech had led them to make this judgement. Of a total of 166 responses 33 related to overall voice quality, 59 to suprasegmental aspects and 74 to segmental aspects. The results of the study are discussed and compared to the literature. PMID- 8241583 TI - Laryngographic analysis of child-directed speech. AB - This paper reports the results of a small-scale study of mother-child interactions using electrolaryngography to measure mean, mode and range of fundamental frequency variation. The mother-child results are compared with those obtained from recordings of the same subjects in conversation with an adult. The methodology employed allowed statistics to be derived from very much larger samples of speech than have hitherto been used in the literature on child directed speech. The main findings were that measures of central tendency are higher for adult-child interactions than for adult-adult conversations and the range of fundamental frequency greater. This expansion of the range is effected by increasing the upper limit of the range. PMID- 8241584 TI - The effects of short courses in modifying the attitudes of adult and adolescent stutterers to communication. AB - Two studies examined attitude changes to communication by stutterers during short intensive courses. The Perceptions of Stuttering Inventory was used to monitor change. As the experimental hypothesis anticipated, change was most clearly seen in the avoidance items of the inventory. In both studies these showed significantly greater positive change than either the struggle or expectancy items. Moreover, positive changes were only found during the period of the course; no positive changes were found for any scales during the month prior to or after the course. The findings justified the use of short courses to change the attitudes of stutterers and suggested that they may be used in conjunction with other approaches which seek to influence fluency directly. An attempt to relate the changes in attitude to changes in speech over the short period of the course was unsuccessful. PMID- 8241585 TI - Paediatric speech and language therapy referral in Singapore: implications for multilingual language disability. AB - Singapore is extremely multilingual, a situation that presents many difficulties for speech and language therapists. A prevalent attitude of tolerance of language use appears to mask language disability in children, and referral rates are low. PMID- 8241586 TI - The prognosis of idiopathic portal hypertension in Japan. AB - To clarify the factors relating to the prognosis of patients with idiopathic portal hypertension (IPH), we followed 171 patients with IPH until the end of 1990, who were registered in the database among those with abnormal portal circulation as of 1985 in hospitals of Japan. During the follow-up period, twenty patients died; 6 from gastro-intestinal tract bleeding, 5 from hepatic insufficiency and 9 from other causes. Cox's proportional hazard model suggested that male patients (hazard ratio 4.85, 95% confidence interval 1.82-12.94), with a disease onset at less than 40 years of age (H.R.3.94, 95% C.I. 1.31-11.57), and/or with varices (H.R.2.86, 95% C.I. 1.05-7.77) generally had poorer prognoses. PMID- 8241587 TI - Distribution of mesenteric and splenic blood flow in cirrhosis of the liver as estimated by radionuclide angiography. AB - We used radionuclide angiography following an injection of 185 MBq Tc-99m tin colloid as a non-invasive method for measuring the mesenteric and portal components of the total hepatic circulation, according to the method of Fleming et al (J Nucl Med 24; 1983). Fifty-seven patients with cirrhosis and 33 controls were examined. The relative mesenteric flow (MF) and the relative portal flow (PF) were 68.6 +/- 8.5% and 78.6 +/- 5.9%, respectively, in the control group. These values were 42.0 +/- 16.4% and 70.9 +/- 8.9%, respectively in the cirrhosis group, and were significantly lower than in the controls. In the cirrhosis group, the reduction in PF was small, while that in MF was large. Consequently, we found that portal blood flow was relatively well maintained in the cirrhosis patients, while the contribution of splanchnic blood flow to the portal flow was relatively reduced. PMID- 8241588 TI - Deterioration of congestive heart failure after converting to VOO mode from DDD mode in a dilated cardiomyopathy patient; importance of atrial contribution. AB - We describe a 62-year-old dilated cardiomyopathy patient in whom congestive heart failure progressed shortly after DDD mode was changed to VOO mode because of battery exhaustion. Emergency battery exchange with restoration of DDD mode improved his functional status to the previous level; pulmonary capillary wedge pressure decreased from 28 to 24 mmHg and cardiac output increased from 1.5 to 2.0 L/min associated with widening of the aortic pulse pressure. Maintenance of atrial contraction is therefore important even if left ventricular systolic function is markedly impaired and left ventricular end-diastolic pressure is increased. PMID- 8241589 TI - Prevalence of gastroesophageal varices supplied by intrahepatic portal branch. AB - The contribution of the intrahepatic portal branch to the gastroesophageal varices is rare. This anomalous pathway (a large left intrahepatic portal branch) was proved by portographic study and by imaging techniques in a 54-year-old man with cirrhosis. The overall prevalence of this type of collateral in the literature is only 1.9% in 908 patients with portal hypertension. In addition, all are via the left portal venous branch. It may be speculated that the mechanism of this variation is due to dilatation of the rest of the intrahepatic portal system via a small anastomosis due to portal hypertension. PMID- 8241590 TI - Chromosomal translocation t(13;16) in a patient with idiopathic hypogonadotropic hypogonadism. AB - A patient with idiopathic hypogonadotropic hypogonadism (IHH) had an apparently balanced reciprocal translocation involving chromosomes 13 and 16 [t(13;16)(q14.11;q24)]. The patient's father has the same chromosomal translocation with no apparent physical abnormalities. The role of the chromosomal translocation in this patient is discussed. PMID- 8241591 TI - Diffuse cerebrospinal gliomatosis mimicking amyotrophic lateral sclerosis. AB - We present a case in which the clinical manifestations mimicked amyotrophic lateral sclerosis (ALS) and the pathological findings showed diffuse cerebrospinal gliomatosis. Diffuse cerebrospinal gliomatosis may be disguised as ALS when it predominantly involves the spinal cord and brain stem, and may not be easily differentiated from ALS. Therefore, diffuse cerebrospinal gliomatosis should be taken into consideration in the differential diagnosis of ALS. PMID- 8241592 TI - Primary pulmonary B-cell lymphoma diagnosed by kappa-lambda imaging of broncho alveolar lavage fluid lymphocytes. AB - A 69-year-old woman was examined due to abnormal pulmonary shadows on her chest roentgenogram. Although small lymphocyte proliferation was revealed in specimens by transbronchial lung biopsy (TBLB), immunoglobulin light chain restriction could not be seen. We attempted to verify the B-cell clonality by broncho alveolar lavage (BAL) and a new sensitive method called kappa-lambda imaging (KLI), which was available for the detection of monoclonal B cells. Therefore, B cell monoclonality was found. Thus, the patient was diagnosed as having primary pulmonary lymphoma (PPL). PPL may be differentiated from benign lymphoproliferative disorders such as pseudolymphoma and lymphocytic interstitial pneumonia by KLI of BAL-derived lymphocytes. PMID- 8241593 TI - Increased RAHA titer and interleukin-6 levels in the synovial fluid in a patient with polymyalgia rheumatica. AB - We report a patient with polymyalgia rheumatica (PMR) accompanied by an increased Rheumatoid Arthritis Hemagglutinin Test (RAHA) titer and interleukin-6 level in the synovial fluid. A 60-year-old female was admitted because of polymyalgia, a body temperature of 39.2 degrees C, and an erythrocyte sedimentation rate increased to 94 mm/h. Since a muscle biopsy failed to show a specific finding, she was diagnosed as PMR. The titer of RAHA and the interleukin-6 level were increased only in the synovial fluid; prednisolone treatment decreased both. The present case raised the possibility that a similar mechanism in rheumatoid arthritis may involve the development of synovitis in PMR. PMID- 8241594 TI - Alcoholic hypertriglyceridemia with decreased activity of lipoprotein lipase and hepatic triglyceride lipase. AB - A 35-year-old male with alcoholic hypertriglyceridemia due to decreased lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) activities is reported. The patient had been drinking about 180 ml of whiskey (equivalent to 80 g of 100% ethanol) every day for the last 17 years, and the highest levels of serum triglyceride (TG) and cholesterol were 5,120 mg/dl and 506 mg/dl, respectively. Serum TG level returned to normal levels after complete alcohol abstinence. Further intake of ethanol resulted in an increase in serum TG to 326 mg/dl with a concomitant decrease in the serum levels of LPL and HTGL activities. PMID- 8241595 TI - Squamous cell carcinoma showing lobar consolidation without collapse. AB - Most squamous cell carcinoma of the lung arises in the large bronchi and causes bronchial obstruction. We report a 74-year-old man with a squamous cell carcinoma of the lung whose roentgenogram revealed a lobar consolidation without collapse. A postmortem examination showed that the tumor cells filled the alveolar spaces completely without destroying the original architecture of the lung. The finding that tumor cells replacing alveolar epithelium, as in bronchioloalveolar cell carcinoma, was not seen throughout the lung. Airway obstruction by the tumor was not observed at any bronchus. These pathological findings seemed to explain the radiological findings. PMID- 8241596 TI - Hepatic cirrhosis showing false-positive serum C-reactive protein reaction. AB - A 58-year-old male who had been diagnosed as hepatic cirrhosis four years previously was admitted to our hospital because his serum C-reactive protein (CRP) level had gradually risen, reaching 139 mg/dl. No inflammation findings were observed subjectively or objectively. Close examination revealed his CRP reaction to be false positive. His serum CRP showed positive only in a latex agglutination method using goat anti-CRP IgG. This false-positive reaction was thought to be owing to the abnormally glycosylated IgM, which has an affinity for the goat serum IgG. PMID- 8241597 TI - Non-Hodgkin's malignant lymphoma of the bone with intracavitary cardiac involvement. AB - An 84-year-old man with non-Hodgkin's malignant lymphoma presented with a pathological fracture of the femur and also an intra-atrial mass on echocardiography. The patient was given palliative low-dose chemotherapy, but died of tumor dissemination, hypercalcemia, and dehydration. Autopsy revealed diffuse large B-cell lymphoma involving multiple bones and lymph nodes (the latter were not detected during staging), and a solitary mass on the posterior wall of the right atrium. This rare lesion appeared to be due to hematogenous spread, since the myocardium and pericardium were free of tumor infiltration. A literature review indicated that the frequency of cardiac involvement differed between contemporary and earlier reports, mainly due to the availability of increasingly sophisticated diagnostic methods and improvements in chemotherapy. PMID- 8241598 TI - Rheumatoid arthritis in a patient with pseudoxanthoma elasticum. AB - Pseudoxanthoma elasticum (PXE) is a rare, inherited disorder of the connective tissue. Possible association of autoimmune thyroiditis and PXE has been suggested, but reports of other autoimmune diseases complicating PXE are rare. We report a case of rheumatoid arthritis (RA) in a patient with PXE. Since the frequency of PXE is likely to be underdiagnosed, further studies to elucidate the true incidence and significance of the association of RA and PXE will be needed. PMID- 8241599 TI - Hypertrophic cranial pachymeningitis associated with mixed connective tissue disease; a comparison with idiopathic and infectious pachymeningitis. AB - This is the first report describing hypertrophic cranial pachymeningitis which developed in association with mixed connective tissue disease (MCTD). A 56-year old man with a two-year history of MCTD gradually developed symptoms of headache and blurred vision. Bilateral papilledema and increased opening pressure and mild pleocytosis of the cerebrospinal fluid were noted. Magnetic resonance imaging of the brain revealed a diffuse thickening and a gadolinium-enhancement of the cranial dura mater. While five previous patients, diagnosed as idiopathic or infectious hypertrophic cranial pachymeningitis in our department, had either otitis media or sinusitis and showed multiple cranial nerve involvement, the present patient had neither otitis media nor sinusitis and did not show any cranial nerve involvement other than papilledema. PMID- 8241600 TI - Disseminated intravascular coagulation in a patient with systemic lupus erythematosus with lupus anticoagulant. AB - A 19-year-old female patient with systemic lupus erythematosus (SLE) who had lupus anticoagulant (LA) and concurrently developed disseminated intravascular coagulation (DIC) during an exacerbation of "central nervous system (CNS) lupus". Since no other indications or causes for DIC could be demonstrated, she was treated with prednisolone and anticoagulants, which rapidly alleviated her DIC condition as well as "CNS lupus". Although abnormalities of coagulation are frequently reported in SLE patients, DIC rarely occurs in patients with SLE without associated complications. A review of the pathogenesis of DIC in patients with SLE is discussed. PMID- 8241601 TI - Skeletal growth factors. AB - Growth factors are polypeptides with important actions on the replication and differentiated function of cells. Skeletal cells synthesize a variety of growth factors, which are believed to act in an autocrine or paracrine fashion. These growth factors include platelet-derived growth factor, fibroblast growth factor 1 and 2, insulin-like growth factor I and II, transforming growth factors beta 1, 2, and 3, and selected bone morphogenetic proteins. Skeletal cells also synthesize specific binding proteins for selected growth factors. In addition, bone marrow cells synthesize a variety of cytokines known to have important actions in bone remodeling. Fibroblast growth factors and platelet-derived growth factors are, for the most part, mitogenic for skeletal cells, whereas insulin like growth factors and transforming growth factor beta enhance the differentiated function of the osteoblast. Growth factors also modify osteoclast recruitment and function and as such, bone resorption. Skeletal growth factors can be regulated at the level of synthesis, activation, binding proteins, and receptor binding, and, as a result, their activity can be modified by exogenous agents. PMID- 8241602 TI - Histone-modulated gene activity: developmental implications. PMID- 8241603 TI - Nuclear localization signals (NLS). AB - Available data strongly suggest that simple karyophilic clusters of arginines and lysines in nucleus-targeted proteins signal the anchoring of these proteins to specialized transporter molecules found on the pore complex or in the cytoplasm. These peptides have been termed nuclear localization signals (NLS). Other nuclear proteins contain "split" or "bipartite" NLS hypothesized to be reconstituted by protein folding or conformational change induced by glucocorticoids. The active NLS needs to be exposed to the protein surface, the cell has invented mechanisms to expose a hidden or cryptic NLS by protein phosphorylation, dephosphorylation, dissociation of an inhibitory subunit that masks the NLS, processing of a larger precursor, and binding of a hormone to regulate the nuclear import of a protein transcription factor at a certain stage of development, or cell cycle. It is proposed (1) that a hexapeptide with four arginines and lysines (and histidines, in some cases) is a good candidate for a "core NLS," (2) that acidic domains on proteins to be imported may participate in anchoring them to the transporter cytoplasmic or pore complex NLS-receptor, and (3) that NLS have both a cytoplasmic and a nuclear function. The interaction between nuclear proteins and transporter proteins in the pore appears to be largely electrostatic and to be disrupted by the binding of mRNA molecules to the same transporter protein, functioning also for the cytoplasmic export of RNA. PMID- 8241604 TI - What constitutes evidence for drug efficacy in osteoporosis? PMID- 8241605 TI - Calcium antagonists in the elderly. AB - The calcium antagonists are associated with a number of advantages over other antihypertensive agents, such as a lack of metabolic, vascular and respiratory adverse events, yet are effective in reducing blood pressure. The currently available calcium antagonists are widely used, and new members, particularly of the dihydropyridine group, continue to emerge. These agents may well be considered for use in the management of hypertension and angina in elderly people. They undergo significant first-pass metabolism and tend to have high values of hepatic clearance with minimal amounts of unchanged drug in the urine. Plasma concentrations tend to be higher in elderly people and for that reason it may be prudent to initiate therapy with lower dosages. With this caveat, adverse effect profiles seem to be qualitatively and quantitatively similar in younger and older people. At equivalent plasma concentrations, the antihypertensive effect appears similar in young and elderly patients, and clinical studies point to comparable efficacy with other drug classes. Calcium antagonists do not have adverse renal, respiratory, cardiovascular, metabolic or peripheral vascular effects and therefore may be useful in patients with relevant concomitant disease. PMID- 8241606 TI - Alterations in neuropeptides in aging and disease. Pathophysiology and potential for clinical intervention. AB - Marked specific and selective changes in the levels of some neuropeptides in age related diseases, such as senile dementia of the Alzheimer (SDAT) or Lewy body (SDLT) types, Parkinson's disease, Huntington's disease and major depressive disorder, versus normal aging have been noted. However, the levels of most neuropeptides are normal. The only 2 peptides consistently altered in SDAT are somatostatin and corticotrophin-releasing hormone both of which are reduced. In Huntington's disease, the level of substance P in the basal ganglia is reduced suggesting a preferential vulnerability of spiny neurones in this disease. In Parkinson's disease, substance P is attenuated in the basal ganglia while somatostatin is reduced in the neocortex. These and other results suggest that substance P deficits are related to movement disorders while somatostatin deficits are related to cognitive impairment. SDLT is a type of dementia with features common to both SDAT and Parkinson's disease, although the changes in neuropeptides suggest that neurochemically the disease is more closely related to SDAT. In major depressive disorder, the level of corticotrophin-releasing hormone is reduced while there is a reciprocal increase in corticotrophin-releasing hormone receptors suggesting that the neurones remain functional. Potential clinical intervention has been limited by problems such as poor penetration of agents into the brain and the short half-lives of neuropeptide agonists and antagonists. However, some currently available agents may act, at least in part, through modulation of neuropeptide pathways, e.g. carbamazepine and alprazolam both modulate the corticotrophin-releasing hormone system in animals, and both have clinically proven antidepressant activity. PMID- 8241607 TI - Drug-induced mania in the elderly. AB - Mania is a rare complication of prescribed drug use in elderly persons. The drugs most likely to produce mania include corticosteroids and dopamine agonists. It is uncertain if antidepressants cause mania or only increase the propensity for latent bipolar mania to be expressed. Drug-induced mania may promote poor judgment, risk-taking behaviours and medical noncompliance. The majority of episodes of drug-induced mania are short-lived and respond to discontinuation of the drug. Antipsychotic agents may hasten resolution of this psychiatric syndrome. Lithium may prevent corticosteroid-induced mania, but the role of lithium in the treatment of other drug-induced manic episodes has not been studied adequately. PMID- 8241610 TI - Drug-induced immune cytopenia. PMID- 8241609 TI - Tiapride. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic potential in geriatric agitation. AB - Tiapride is a substituted benzamide derivative with selective dopamine D2 receptor antagonist properties which appears to have preferential affinity for extrastriatal dopamine receptors. Animal and clinical studies show that tiapride has anxiolytic properties but the mechanism of action is uncertain. Results from limited studies indicate that the clinical efficacy of tiapride in the treatment of agitation, aggressiveness, anxiety and sleep disorders in the elderly appears superior to that of placebo, chlorpromazine, lorazepam and meprobamate. Tiapride also exerts a beneficial effect on vigilance and alertness in elderly patients and causes less sedation than chlorpromazine. Tiapride is well tolerated at the dosages recommended for elderly patients. Further well designed comparative studies with newer drugs are needed to determine the relative place of tiapride in the treatment of geriatric agitation, and such studies should also address the quality-of-life benefits for the patient. Additional clinical experience to determine the efficacy of tiapride in elderly patients with more than one disease condition, receiving concomitant medications, and/or with renal impairment is also required. However, despite these current limitations, tiapride may have potentially important applications in this difficult area of clinical medicine. PMID- 8241608 TI - Zopiclone. A review of its pharmacological properties and therapeutic efficacy as an hypnotic. AB - Zopiclone is a cyclopyrrolone which is chemically unrelated to the benzodiazepines and is thought to act on the GABAA receptor complex at a site distinct from, but closely related to, the benzodiazepine binding site. The hypnotic efficacy of zopiclone administered as single oral doses has been demonstrated in patients undergoing next-day surgery and in patients with insomnia, and these studies have established an optimal dose of 7.5mg for elderly patients. Using this dose, clinical studies have shown that zopiclone improved sleep in elderly patients to a similar extent as triazolam 0.125 to 0.5mg, flurazepam 15mg, and nitrazepam 5mg. Studies that also included younger patients have shown that zopiclone 7.5mg is at least as effective as triazolam 0.25 or 0.5mg, and on most sleep parameters is comparable to temazepam 20mg, nitrazepam 5mg, flunitrazepam 2mg, and flurazepam 20mg. Zopiclone causes minimal impairment to psychomotor performance and mental alertness the morning after night-time administration. The drug is generally well tolerated by patients of all ages; the most frequently reported adverse effects being bitter taste and dry mouth. Treatment withdrawal due to adverse effects is seldom required and reports of rebound insomnia after zopiclone withdrawal are rare. While symptoms of physical dependence have not been observed in clinical studies, there have been isolated reports of physical dependence in patients with a history of substance abuse. Although the latter finding should be kept in mind, it appears that zopiclone has a low dependence liability. Thus, with its short duration of action and good tolerability profile, zopiclone is a well established alternative to the benzodiazepine hypnotics and may be particularly beneficial in those patients unable or unwilling to tolerate the residual effects associated with many other hypnotic agents. PMID- 8241611 TI - Mechanisms of drug-induced immunologically mediated cytopenias. PMID- 8241612 TI - Specificity of drug-induced immune cytopenias. AB - What conclusions can be drawn concerning specificity of drug-induced immune reactions? We have seen that specificity of these reactions depends on several molecular features including the chemical nature of the drug, specific domains of particular membrane components, and as yet unidentified characteristics that determine selectivity for one or more cell types. This latter property does not seem to be related to shared membrane components because, for example, Rh antigens on RBCs, the peptide tail region of GPIb alpha on platelets, and the 85 kd GP on neutrophils are clearly not part of the same molecules. From multiple studies of quinine/quinidine-dependent and nomifensine-dependent antibody interactions with platelets and RBCs, respectively, we can conclude that these particular reactions are a function of specific features of the drug molecules and specific domains of various membrane glycoproteins. These characteristics strongly argue that the hypervariable regions of drug-dependent, platelet and RBC antibodies recognize simultaneously a specific domain of the membrane GP and a specific configuration of the drug molecule. If this is true, then it follows that together a specific domain of the cell membrane component plus the drug define an antigenic determinant or epitope for attachment of certain drug dependent antibodies. We have also seen that some drug-dependent antibodies preferentially react with the drug alone when it is attached to cell membranes (eg, penicillin-dependent antibodies reacting with penicillin-coated RBCs or platelets). Some drugs elicit antibodies that react at specific sites on the cell membrane independently of drug (eg, nomifensine and the Rh antigens (E) or quinidine and platelet GPV). These three concepts of antibody specificity induced by drugs are presented in Fig 6, using RBCs as an example. Despite major advances in understanding drug-induced immune reactions during the past four decades, several important questions remain to be answered. For example, why are platelets involved more frequently than other cells of the circulation in these types of reactions? Why do some individuals develop drug-induced immune cytopenias that are specific for a single cell type, whereas others develop reactions involving multiple cell types with distinct antibodies? What mechanism directs the reaction toward platelets, RBCs, or neutrophils? How are drug-dependent antigens presented to the immune system? This latter question is particularly intriguing considering that most drugs known to induce immune cytopenias bind only weakly to target tissues.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8241613 TI - Drug-induced autoimmune hemolytic anemia. PMID- 8241614 TI - Immune cytopenia associated with antibiotics. PMID- 8241615 TI - Drug-induced immune neutropenia. AB - Drug-induced immune-mediated neutropenia is a serious complication of therapy with many different drugs. Its clinical presentation and outcome are highly variable. In addition to the drugs listed in this review it is likely that many other drugs also cause immune-mediated neutropenia but drug-dependent neutrophil antibodies were not reported because of the lack of widespread availability of testing. With the exception of quinine-dependent antibodies in three patients, the neutrophil antigens recognized by the drug-dependent antibodies are not known; however, the recent development of MAIGA assays will likely help identify the antigens other drug-dependent neutrophil antibodies recognize. PMID- 8241616 TI - Laboratory investigation of drug-induced immune thrombocytopenias. PMID- 8241617 TI - 137cesium and 134cesium in roe deer from north and middle Hesse (Germany) subsequent to the reactor accident in Chernobyl. PMID- 8241618 TI - Distribution of some organochlorine compounds (PCB, CBz, and DDE) in beeswax and honey. PMID- 8241619 TI - Resistance of lactogenic microorganisms and mixed ruminal bacteria to pentachlorobiphenyl (Delor 105). PMID- 8241620 TI - Pollution studies in the Rio Santiago Basin, tributary of the Rio de la Plata Estuary: preliminary risk assessment evaluation. PMID- 8241621 TI - Cadmium, lead, and zinc from terrestial plants in the Enyigba-Abakaliki lead and zinc mine: search for a monitoring plant species in trace element distribution. PMID- 8241622 TI - Distribution of inorganic compounds in sediments from three waterways in northern New Jersey. PMID- 8241623 TI - Structure-toxicity relationships for unsaturated alcohols to Tetrahymena pyriformis: C5 and C6 analogs and primary propargylic alcohols. PMID- 8241624 TI - Naphthalene-induced cytotoxicity on the hepatopancreatic cells of the red swamp crayfish, Procambarus clarkii. PMID- 8241625 TI - Acute toxicity of several pesticides, organic compounds, and a wastewater effluent to the freshwater mussel, Anodonta imbecilis, Ceriodaphnia dubia, and Pimephales promelas. PMID- 8241626 TI - Effect of sublethal concentrations of glyphosate and dalapon on protein and aminotransferase activity in Pseudosuccinea columella. PMID- 8241627 TI - Mercury concentration of stickleback Gasterosteus aculeatus from the Gulf of Gdansk. PMID- 8241628 TI - Salinity tolerance and growth analysis of the cyanobacterium Anabaena doliolum. PMID- 8241629 TI - Assessment of certain hematological responses of factory workers exposed to pesticides. PMID- 8241630 TI - Effects of 2-chlorodibenzofuran on fetal development in mice. PMID- 8241631 TI - Selenium metabolite levels in human urine after dosing selenium in different chemical forms. PMID- 8241632 TI - Differential accumulation of thallous ion by diverse rabbit and rat muscles. PMID- 8241633 TI - Lipid peroxidation induced by maternal cadmium exposure in mouse pups. PMID- 8241634 TI - The implications of recent advances in coeliac disease. PMID- 8241635 TI - Serum selenium levels in neonatal hyperbilirubinemia. PMID- 8241636 TI - Influence of dietary taurine on vitamin D absorption. AB - To evaluate the influence of dietary taurine supplementation on vitamin D absorption, we studied three groups of infants: 21 (11 preterm) were fed a taurine-free formula, 21 (10 preterm) were fed a taurine-supplemented formula (50 mg/100 g of powder) and 20 (9 preterm) were fed human, not heat-treated milk. Taurine, total bile acids, glyco-(GBA) and tauro-(TBA) conjugated bile acids, 25 hydroxyvitamin D3 (25OHD3) and 1,25-dihydroxyvitamin D3 (1,25OH2D3) were determined in all infants at birth in blood cord and at one and three months of life. In preterm infants fed a taurine-free formula, we found lower plasma taurine levels than in infants of other groups at one and three months of life. In these infants, GBA predominated, with a G/T ratio of 1.1 and 1.4 at one and three months of life, whereas in all other infants TBA predominated with a G/T ratio always < 1. Also, 25OHD3 and 1,25OH2D3 levels were significantly lower in preterm infants fed a taurine-free formula than in infants fed a taurine-enriched formula or human milk. Term infants fed a taurine-free formula did not show differences in the parameters studied in comparison to infants of other groups. Low taurine dietary intake appears to compromise vitamin D absorption in preterm infants, and therefore taurine supplementation of preterm infant formulas should be encouraged. PMID- 8241637 TI - Feeding and nutritional problems in children with cerebral palsy and myelomeningocoele. AB - This study was undertaken to characterize the food intake, current feeding situation and nutritional status of children with cerebral palsy and myelomeningocoele. Thirty children with cerebral palsy and 14 with myelomeningocoele were investigated. The children with cerebral palsy were subgrouped into those with diplegia (11 children), dystonia (10 children), hemiplegia (6 children) and ataxia (3 children). Information was obtained from medical records, current dietary recording and clinical and anthropometric examinations. Feeding problems, low energy intake and wasting were frequent among the children with cerebral palsy, particularly those with the dystonic form. Children with myelomeningocoele gave the impression of being overnourished, despite a seemingly relatively low energy intake. It is clear that nutritional and feeding aspects should receive particular attention in the habilitation of children with various disabilities. PMID- 8241638 TI - Osteopontin in human milk from mothers of premature infants. PMID- 8241639 TI - Histological findings and evidence of lipid conjugated dienes and malonyldialdehyde in human fetal aortas. AB - Recent evidence strongly suggests that peroxidative modification of lipids may play a significant role in atherogenesis. In our present research, we investigated if the oxidative stress mediated by oxygen free radicals was a pathophysiologic condition that occurred in the early stages of human development. Thus the aim of this research was to examine lipid peroxidation in human fetal aortas. Human fetal aortas and proximal iliac arteries (n = 8) were obtained from fetuses aged 7 +/- 2 months, immediately after autopsy. Lipids from the initial fatty streak lesions (LFS) and the vessels uninvolved (LUV) were extracted by the chloroform/methanol method. Lipid peroxidation levels were measured by two different methods: determination of lipid conjugate dienes (the spectrum trend was recorded from 320 to 200 nm with a spectrophotometer) and malonyldialdehyde (MDA) content (TBA method). We observed that lipid conjugated dienes were present in LFS, but not in LUV, with a characteristic absorption peak at 233 nm. In addition, MDA levels were significantly higher when the LFS = 3.85 +/- 0.91 nmol than when the LUV = 0.41 +/- 0.12 nmol (p < 0.001 versus LUV). The presence of lipid peroxidation in our samples could be mediated by free radical production in the first stages of human development. Thus these data suggest that LFS peroxidation mediated by free radicals occurs in the vascular circulation in the early stages of human development. This could influence the progression of vascular damage and atherosclerotic disease. PMID- 8241640 TI - Perinatal factors associated with retinopathy of prematurity. AB - The etiology of retinopathy of prematurity appears to be multifactorial. Introduction of new treatments in neonatal care may add new risk factors. We have analyzed the relationship between 42 perinatal factors and the development of retinopathy of prematurity in 78 infants with a birth weight < 1501 g and/or gestational age < 33 weeks. We have also applied a chronological analysis of the maximum and minimum pO2 and pCO2 values. Retinopathy of prematurity was seen in 37 of 78 infants (47.4%). Nineteen factors were found to be related to the development of retinopathy of prematurity. However, when step-wise logistic regression analysis was used, only birth weight, number of days of oxygen therapy and use of beta-blocking agents by the mother before birth were found to be associated with the development of retinopathy of prematurity. The results suggest that medication with beta blockers immediately before birth should be used cautiously. PMID- 8241641 TI - Protection against Campylobacter diarrhea: role of milk IgA antibodies against bacterial surface antigens. AB - In developing countries, Campylobacter jejuni causes diarrhea and dysentery, especially in children less than one year of age. Breast feeding protects against infectious diarrhea, with milk IgA antibody playing a determining role. Therefore, it has been proposed to increase the protective effect of human milk by vaccinating women of child-bearing age. To identify antigens which may induce protective breast-milk IgA, we analyzed 60 strains of C. jejuni isolated from asymptomatically- and symptomatically-infected breast-fed children less than 12 months of age. Surface antigens of C. jejuni, separated by polyacrylamide gel electrophoresis, were probed with breast milk collected concurrently with the fecal sample from which C. jejuni was isolated, and specific IgA was developed by immunoblotting. Our results indicate that milk antibodies against three high molecular weight bacterial surface antigens of 95, 110 and 185 kDa are involved in protection of infants infected with C. jejuni (p = 0.00964 for one-tailed Fisher's exact test). PMID- 8241642 TI - Prolonged pure granulocytopenia in children. AB - In a prospective, population-based study, 31 patients with pure granulocytopenia lasting longer than six months were follow-up for up to 7.3 years after diagnosis. Their ages at diagnosis were 0.3-15.5 (median 0.9) years; 17 of the patients were less than 1 year of age. The lowest granulocyte count measured was 0.00-0.60 (median 0.03) x 10(9)/l. During the granulocytopenia, 12 patients suffered from repeated infections but none was life-threatening. In 21 patients, the granulocyte count normalized spontaneously within 0.5-5.5 (median 1.1) years: the other 10 remain granulocytopenic after follow-up for 2.5-7.3 (median 3.7) years. We conclude that selective granulocytopenia, if associated with unimpaired myelopoiesis, is a benign disorder in children and does not, even if prolonged, significantly increase the risk of severe infections. PMID- 8241643 TI - Prevalence of potential respiratory disease bacteria in children in Ethiopia. Antimicrobial susceptibility of the pathogens and use of antibiotics among the children. AB - Acute respiratory infections are primary causes of morbidity and mortality in children in developing countries. This project was designed to investigate antimicrobial susceptibility of respiratory tract pathogens isolated from children in rural and city areas, and to contribute to the rational choice of antibiotics for respiratory tract infections in children in Ethiopia. Nasopharynx and throat cultures were taken from all children under five years of age in three study areas representing different levels of contact with health care and accessibility to modern drugs, such as antibiotics. In all, 1126 children were cultured. Haemophilus influenzae and Streptococcus pneumoniae were both found in 85-90% of the children, and beta-haemolytic streptococci group A in 12%. The level of antimicrobial resistance was low. None of the 954 strains of H. influenzae were beta-lactamase producers. Pneumococci were susceptible to penicillin. The use of antibiotics was also low; 11 of 1126 children had antibiotics on the day of culture or the day before. The choice of antibiotics was not limited by resistance, and emphasis could be put on low cost, minimizing adverse drug reactions and ecological impact. PMID- 8241644 TI - Concentrations of nucleotides, nucleosides, purine bases and urate in cerebrospinal fluid of children with meningitis. AB - The release of agents mediating inflammation in meningitis may bring about neuronal hypoxia, under which circumstances ATP concentrations decrease and its degradation products increase and are released into the cerebrospinal fluid. In this study of alterations in neuronal energy metabolism in meningitis, AMP, IMP, inosine, adenosine, guanosine, adenine, guanine, hypoxanthine, xanthine and urate were determined by high performance liquid chromatography in the cerebrospinal fluid of 54 children aged between 1 month and 13 years suffering from meningitis (25 viral, 24 bacterial and 5 tuberculous cases) and 63 controls. Compared to the controls, patients with viral meningitis exhibited high concentrations of IMP, adenosine, guanosine, adenine, guanine and xanthine; patients with bacterial meningitis exhibited high concentrations of IMP, inosine, guanosine, adenosine, hypoxanthine, xanthine and urate; and patients with tuberculous meningitis exhibited high concentrations of AMP, guanosine, xanthine and urate. Viral and bacterial cases did not differ significantly for any of the metabolites studied. AMP and urate concentrations were significantly higher in patients with tuberculous cases compared with viral or bacterial meningitis cases. PMID- 8241645 TI - Long-term treatment with growth hormone in Noonan's syndrome. AB - Five children (four boys, one girl) with Noonan's syndrome were treated with growth hormone for a mean of 2.9 (range 1.8 to 4.6) years. Treatment was commenced at a mean age of 3.9 (range 2.5 to 6.0) years. After one year of treatment, height velocity SDS increased from a mean of -2.1 (range -4.1 to -0.3) to a mean of +3.1 (range +2.0 to +3.5). Subsequently, height SDS increased significantly from -3.3 (range -4.2 to -2.2) to -2.4 (range -3.3 to -1.6). Although in all patients a marked decrease in height velocity was observed in the following years of treatment, compared to the first year, height SDS continued to improve in comparison to pretreatment (mean height SDS in the third year for four patients was -2.1) (range -3.1 to -1.4). In the four patients who had completed their second year of treatment, height SDS for bone age had increased from -1.8 (range -2.8 to -0.9) to -1.2 (range -1.6 to -0.6). PMID- 8241646 TI - Early treatment with nasal-CPAP. PMID- 8241647 TI - Urinary albumin excretion rate and puberty in non-diabetic children and adolescents. AB - Slightly elevated urinary albumin excretion rate (microalbuminuria) is a marker of early diabetic nephropathy, but it is unclear if the established definition of microalbuminuria (20-200 micrograms/min) is correct for children and adolescents. We investigated the albumin excretion rate, albumin/creatinine ratio and urinary albumin concentration in 150 healthy schoolchildren and adolescents to (a) obtain a reference value for albumin excretion rate, (b) relate albumin excretion to pubertal stages and (c) evaluate albumin/creatinine ratio and morning albumin concentration as screening methods for elevated albumin excretion rate. Albumin concentration was measured by immunoturbidimetry in timed overnight urine samples. The albumin excretion showed a skewed distribution (geometric mean 3.2 micrograms/min, 95 percentile 15.1 micrograms/min). In girls, a peak in the albumin excretion rate was found at the pubertal stage 4 (Tanner) and in boys at stage 5. Albumin/creatinine ratio of 2.5 mg/mmol as a screening level for elevated albumin excretion (15 micrograms/min) showed a high positive (0.88) and negative (0.99) predictive value. PMID- 8241648 TI - Obstructed breathing in children during sleep monitored by echocardiography. AB - Six 3 to 14-year-old boys with snoring and obstructive sleep apnea syndrome were monitored polygraphically during sleep with and without nasal continuous positive airway pressure with simultaneous recording of esophageal pressure (Pes) and M mode and two-dimensional echocardiograms. Continuous non-invasive blood pressure monitoring was performed in two older children. Three of the six children demonstrated a diastolic leftward shift of the interventricular septum related to the negativity of Pes. Progressively more negative Pes correlated significantly with an increase in right ventricular internal end-diastolic dimension and a decrease in left ventricular internal end-diastolic dimension, with at times left ventricular "collapse". One of the subjects with blood pressure monitoring demonstrated pulsus paradoxus with leftward shift of the interventricular septum. Nasal continuous positive airway pressure normalized all changes. Pulsus paradoxus and leftward shift of the interventricular septum are related to the mechanical changes associated with heavy snoring during sleep, regardless of the amount of oxygen desaturation. PMID- 8241649 TI - Biometric method for the ossification evaluation of children from birth up to the ages of two and four--applied to the tarsus. PMID- 8241650 TI - Recurrence of unexpected infant death. AB - Families which had experienced two or more unexpected infant deaths were the subject of detailed confidential enquiries, including necropsy examination. Cases were derived from two main sources: first, deaths occurring during a nationwide programme of support for families with a subsequent baby (8 families) plus 2 families from a series of confidential enquiries in Sheffield, and second, direct referrals from paediatricians (17 families). Fifty-seven deaths were studied. Twenty-four families had experienced 2 and three had experienced 3 deaths; 11 deaths (19%) were found to be adequately explained by history or post-mortem findings; 7 (12%) were probably accidental; 31 (55%) were most probably due to an action by one of the parents (filicide); only 5 (9%) were considered to be true or idiopathic sudden infant death syndrome; in 3 (5%) cases there was insufficient information to draw a conclusion. Five (18%) of the families lived in circumstances of serious social deprivation. A history of psychiatric illness was present in one or both parents in 18 (67%) of the families. PMID- 8241651 TI - Acute accidental overdosage of haloperidol in children. AB - We report the details of an accidental overdosage of haloperidol in 24 children in one hospital in Kyushu, Japan. Evidence of acute toxicity included disturbances in consciousness (24/24), tremors in the extremities (16/24), an oculogyric or similar crisis (14/24), dysarthria (9/17), drooling (8/24), akathisia (6/20), hyperreflexia (6/24) and opisthotonos (3/24). Laboratory examinations revealed late-onset transient thrombocytosis (5/24), elevated AST and GPT (1/24) and abnormal ECG with prolonged QT interval in 2 of 8 children. We detected haloperidol in 11 of 18 children whose blood was specifically examined within four days after the final haloperidol administration. The maximum serum haloperidol level was 28.9 ng/ml. The mean half-life of haloperidol in the serum of five children (age range 2-10 years) was 18.6 +/- 12.2 h (mean +/- SD) (range 9.1-39.4 h). PMID- 8241652 TI - Severe hyponatremia caused by intranasal desmopressin for nocturnal enuresis. PMID- 8241653 TI - Oral administration of arginine enhances the growth hormone response to growth hormone releasing hormone in short children. AB - We have evaluated the effect of oral administration of arginine chlorhydrate on the growth hormone response to growth hormone releasing hormone in a group of nine short prepubertal children (six boys and four girls). Arginine chlorhydrate 10 g, administered orally 60 min before an i.v. bolus injection of growth hormone releasing hormone 1-29, 1 microgram/kg, significantly enhanced the growth hormone response to the neuropeptide, confirming the results of previous studies which used the i.v. route. Furthermore, our data strengthen the view that the effects of arginine chlorhydrate on growth hormone secretion are mediated by inhibition of endogenous somatostatin release. PMID- 8241654 TI - Hallucinations on withdrawal of isoflurane used as sedation. AB - Hallucinations on withdrawal of midazolam have been noted in intensive care patients. Isoflurane is increasingly used as an alternative drug for sedation. We report a seven-year-old child with 15% burns who, on withdrawal of isoflurane, suffered hallucinations, a generalized seizure and disorientation. The adverse symptoms occurred within hours of stopping the isoflurane and lasted for five days. PMID- 8241655 TI - Very late mucosal relapse in a girl with coeliac disease. AB - Small bowel biopsy in a 4-year-old girl with symptoms suggestive of coeliac disease revealed subtotal villous atrophy. The mucosa healed on a gluten-free diet. From the age of 7 years, the girl was challenged with gluten. Annual biopsies showed normal or nearly normal mucosa specimens. At 21 years of age, after 14 years of gluten challenge, a mucosal relapse was found and a gluten-free diet was reinstituted. A biopsy one year later showed a normal mucosa. From this case report it is apparent that a patient with a past history of subtotal villous atrophy, which after a preceding period of gluten-free diet does not recur within two years of gluten challenge, must be followed carefully, so as not to miss a late relapse. PMID- 8241656 TI - Cholelithiasis in cervico-oculo-acoustic (Wildervanck's) syndrome. AB - An unusual case of cholelithiasis in an 18-month-old boy with cervico-oculo acoustic (Wildervanck's) syndrome is presented. Our patient had Duane's retraction syndrome, Klippel-Feil anomaly and congenital deafness. To our knowledge this is the first case in which a probable association between cholelithiasis and Wildervanck's syndrome has been recorded. On the other hand, the presence of mutual malformations and anomalies such as scoliosis, ventricular septal defect, ectopic kidney, hydrocephalus, hypoplastic thumb and growth retardation seems to suggest that Wildervanck's syndrome is a clinical variant of Klippel-Feil sequence. PMID- 8241657 TI - Lipoid proteinosis: Urbach-Wiethe disease. AB - Lipoid proteinosis (Urbach-Wiethe disease) is a rare autosomal recessive disorder in which hyalinized material is deposited in the skin, mucous membranes and brain. Laryngeal changes resulting in hoarseness may be present at birth or in early infancy, often being the first sign of the disease. A typical case is presented. PMID- 8241658 TI - Early child health in Lahore, Pakistan. PMID- 8241659 TI - Checklist for the methods section of dietary investigations. PMID- 8241660 TI - Early CPAP/minimal handling. PMID- 8241661 TI - Hypoallergenic formulae. The Executive Committee of ESPACI. PMID- 8241662 TI - Assessment of bone mineral content in infants: the new age. PMID- 8241663 TI - Is there an impact of birth weight and early life nutrition on bone mineral content in preterm born infants and children? PMID- 8241664 TI - Renal function in sick, very low-birth-weight infants. AB - Renal immaturity is pronounced in very low-birth-weight infants with a gestational age < or = 30 weeks. We attempted to elucidate if conditions requiring mechanical ventilation, including patent ductus arteriosus, might further compromise renal function due to decreased renal perfusion. Forty infants studied between 4 and 28 days of age were divided into four groups: Control with no patent ductus or mechanical ventilation (n = 8); PDA+MV, with both patent ductus and mechanical ventilation (n = 17); PDA, with patent ductus (n = 6); MV, with mechanical ventilation (n = 9). The groups PDA+MV and MV had significantly lower creatinine clearances and significantly higher fractional sodium excretions than controls. Mean arterial pressure was significantly lower in all groups compared to controls and correlated significantly with creatinine clearance (r = 0.47, p < 0.02). In conclusion, low renal function in these infants is further compromised by a patent ductus arteriosus and/or the use of mechanical ventilation. PMID- 8241665 TI - Ambient light, ophthalmic artery blood flow velocities and retinopathy of prematurity. AB - Exposure to bright light was recently proposed as a possible risk factor for the development of retinopathy of prematurity. A semi-longitudinal Doppler ultrasound study was conducted of ophthalmic artery flow velocities at normal and increased ambient light in 22 preterm infants, at post-menstrual ages from 28 to 37 weeks. The aim of this study was to obtain relationships between ophthalmic artery blood flow velocities at various post-menstrual ages and lighting conditions and the occurrence of retinopathy of prematurity. A gradual increase in average blood flow velocities was seen between 28 and 37 weeks. A stepwise increase in flow velocity was seen in all cases when ambient light was increased from moderate to intense. Five of the 22 infants developed retinopathy. No association could be established between ophthalmic artery flow velocities or light-induced changes in flow velocity and the occurrence of retinopathy. PMID- 8241666 TI - Personality and behaviour in eight-year-old, non-handicapped children with birth weight under 1500 g. AB - Personality and behavioural characteristics of a population-based cohort of 29 very low-birth-weight (VLBW) infants were compared with those of 29 matched, term controls at eight years of age. The VLBW infants were born to families of lower socioeconomic status (p = 0.04) and had a lower mean IQ (93 versus 104, p = 0.008) and motor abilities (p = 0.028). Based on the questionnaire personality inventory for children, the VLBW children had more learning difficulties and school coping problems, and the VLBW boys had more conduct and emotional problems than the controls. Except for conduct problems, these differences persisted after controlling for socioeconomic status. Generally, there were significant relationships between behaviour, IQ and motor abilities. In conclusion, VLBW may be a risk factor for the development of school coping and behavioural problems independent of socioeconomic status but often coexist with impaired cognitive and neuromotor function. PMID- 8241667 TI - Variation in serum C-reactive protein across the clinical spectrum of meningococcal disease. AB - In a multicentre prospective study, 124 cases of meningococcal disease were classified into the clinical categories, meningitis alone (n = 15), meningitis and septicaemia (n = 79) and septicaemia alone (n = 30). A further 60 children referred with other illnesses served as controls. Serial measurements of serum C reactive protein (admission, day 1, day 2, days 5-7) were compared. Children with septicaemia had significantly lower C-reactive protein levels on admission than those with meningitis alone or meningitis and septicaemia which were unexplained by differences in the duration of the presenting illness or severity of the disease. Within each clinical category of meningococcal disease, significant changes in C-reactive protein concentration occurred during the course of the disease. Four control children had other types of septic meningitis: admission C reactive protein concentrations did not differ from those with meningitis or meningitis and septicaemia, but were significantly higher than those with septicaemia alone. The other 56 patients had a significantly lower admission C reactive protein concentration compared with all cases of meningococcal disease. For the diagnosis of meningococcal disease, admission C-reactive protein levels of > or = 40 mg/l had a sensitivity of 79%, specificity of 80% and positive predictive value of 87%. For the prognostic prediction of death in meningococcal disease (or meningococcal disease with shock) CRP < 100 mg/l on admission had a sensitivity of 69% (69%), specificity of 50% (56%) and positive predictive value of 18% (53%). In children with suspected meningococcal disease, serum C-reactive protein, measured on admission, has diagnostic value but not prognostic value.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241668 TI - Zinc supplementation impairs monocyte function. AB - Zinc has been shown to be involved in many functions of the immune system. This study was conducted to examine the effect of zinc supplementation on phagocytic, fungicidal and metabolic activity of blood monocytes of marasmic infants during nutritional rehabilitation. A controlled, double-blind design was used in which 19 infants fed a zinc-fortified formula were compared with 20 infants fed the same, unfortified formula. Evaluation of phagocytic-fungicidal capacity, growth, zinc, copper and iron status was performed in both groups on admission and after 60 and 105 days of nutritional rehabilitation. Although energy, copper and iron intakes were similar in the two groups, a decrease in the number of infants able to phagocytose one or more Candida buds was observed after 60 days of zinc supplementation compared to admission (p < 0.03). No change in phagocytic ability was detected between admission and 60 days in the control group. The number of infants with depressed fungicidal activity increased significantly after 105 days of nutritional rehabilitation in the zinc-fortified group as compared to controls (p < 0.04). The number and duration of impetigo episodes was significantly greater in the group fed the zinc-fortified formula. These results suggest that zinc supplements at the RDA level may impair monocyte function. PMID- 8241669 TI - Kinetics of hypogammaglobulinemia in patients with common variable immunodeficiency. AB - We investigated the production of immunoglobulin (Ig) in six patients with common variable immunodeficiency and analyzed the courses of their levels of serum Ig for a period of 8-13 years. In all patients, levels of serum IgM, IgG and IgA were markedly low at the first examination, except for the IgM level in one patient. Improvement of serum Ig levels was observed in the patients in whom Ig production of non-T cells at the first examination and C mu gene expression had been detected to a slight degree, but serum Ig levels remained low in the patients in whom Ig production of non-T cells and C mu gene expression had not been detected. Our results suggest that some hypogammaglobulinemia in common variable immunodeficiency develops or improves with age. PMID- 8241670 TI - Hereditary properdin deficiency in three families of Tunisian Jews. AB - Hereditary properdin deficiency is a rare genetic disorder of the complement system. Three propositi and six additional family members with properdin deficiency have been found following analysis of the hemolytic activity of the classical (CH50) and the alternative (AP50) complement pathways in the sera of 101 survivors of meningococcal infections and 59 survivors of severe pneumococcal and Haemophilus influenza infections. All the properdin-deficient individuals had undetectable levels of properdin by radial immunodiffusion and by Western blotting. They belonged to three non-related families of Tunisian Jews who came from different parts of Tunisia. Two patients had a meningococcal infection at 15 and 16 years of age, respectively, and one had Haemophilus influenza meningitis at 1.5 years of age. In contrast to the fulminant and fatal course of meningococcal infection which was previously described in some properdin deficient patients, our patients had a relatively mild disease. Properdin deficiency may not be as rare as previously thought. Analysis of AP50, in addition to CH50, in sera of patients who had meningococcal infection, will probably disclose many more cases of hereditary properdin deficiency. In addition, our findings indicate that, as in other complement abnormalities, hereditary properdin deficiency may also be associated with the ethnic origin of the patient. PMID- 8241671 TI - Prevalence of coeliac disease in diabetic children and adolescents in Sweden. AB - The aim of this study was to determine the minimum prevalence of coeliac disease in a group of 459 diabetic children and adolescents. Six patients were already known to have coeliac disease. A total of 436 patients with type 1 diabetes mellitus aged 2-21 years and with age at onset at 2 months to 17 years at three paediatric departments agreed to participate in the study. All patients were tested for gliadin IgA antibodies with a commercial kit (Pharmacia Gluten IgA EIA). Later, serum was tested for reticulin IgA/IgG antibodies. Nineteen patients had elevated gliadin IgA levels (> 25 AU). Eighteen underwent jejunal biopsy. Ten had total or subtotal villous atrophy. These 10 patients were reticulin IgA positive. Of 417 gliadin IgA-negative patients, 408 were reticulin IgA/IgG negative. Of 6 reticulin IgA-positive patients, 3 had total or subtotal villous atrophy. All 3 had become gliadin IgA-positive at the time of biopsy. Among 3 reticulin IgG-positive patients with IgA deficiency, 2 had total villous atrophy: 1 was not willing to be biopsied. Patients with total or subtotal villous atrophy were judged as having coeliac disease and were recommended a gluten-free diet. Within 2 months, gliadin IgA levels were normal in patients adhering to the diet. Five patients have gone through a second jejunal biopsy to date with normal histology in all 5. The 15 newly diagnosed patients with coeliac disease plus 6 already known patients with coeliac disease and type 1 diabetes mellitus gave a minimum prevalence of coeliac disease in diabetic children and adolescents of 21/459 = 4.6%. PMID- 8241672 TI - Water-balance hormones during long-term follow-up of oral dDAVP treatment in diabetes insipidus. AB - The aim of the present study was to investigate the hormonal control of water balance in children with diabetes insipidus and to assess safety and efficacy of long-term treatment with oral dDAVP. Plasma atrial natriuretic peptide, plasma renin activity, aldosterone, plasma and urinary cyclic 3'5'-guanosine monophosphate and urinary prostaglandin E2 were measured in eight patients (aged 3-21 y) with central diabetes insipidus. At baseline, 12 h after the last dDAVP dose, patients had hypotonic polyuria but normal plasma sodium concentrations and plasma osmolality relative to a control group. The mean plasma atrial natriuretic peptide concentration in patients (26.2 +/- 2.6 pg/ml) tended to be lower than in controls (36.5 +/- 8.2 pg/ml, mean +/- SEM), although the difference was not significant. Plasma cyclic 3'5' guanosine monophosphate was higher in controls (6.0 +/- 0.6 pmol/ml, mean +/- SEM) than in patients (3.8 +/- 0.3 pmol/ml). Aldosterone, plasma renin activity, urinary cyclic guanosine monophosphate and urinary prostaglandin E2 were similar in the two groups. During 3 h following dDAVP administration, atrial natriuretic peptide levels did not change in patients but decreased significantly in controls to 23.0 +/- 4.0 pg/ml. No adverse reactions, or circulating antibodies against dDAVP, were observed after 3.5 years of oral dDAVP treatment. The average oral dDAVP dosage was similar after 1 and 3.5 years of treatment (906 +/- 406 micrograms/24 h, mean +/- SD). Water-balance is not detectably different from normal in correctly treated diabetes insipidus patients in terms of plasma atrial natriuretic peptide, plasma renin activity and aldosterone levels. Long-term oral dDAVP treatment is safe and efficacious. PMID- 8241673 TI - A missense mutation in the hypoxanthine phosphoribosyltransferase gene in a pediatric patient with hyperuricemia. AB - We have identified a mutation in the gene coding for the enzyme hypoxanthine phosphoribosyltransferase in a pediatric patient with hyperuricemia and nephrolithiasis. The mutation is a nucleotide substitution causing an amino acid substitution in the hypoxanthine phosphoribosyltransferase protein. In this patient, fibroblasts but not lymphocytes showed resistance to 6-thioguanine, and reduced enzyme activity was detected in lymphocytes. These results are consistent with the intermediary phenotype associated with partial hypoxanthine phosphoribosyltransferase enzyme deficiency. Altogether, six males in this family suffered from hyperuricemic symptoms, and small differences in phenotype were seen. PMID- 8241674 TI - Serum concentrations of procollagen I C-terminal propeptide, osteocalcin and insulin-like growth factor-I in patients with non-lethal osteogenesis imperfecta. AB - Serum concentrations of procollagen I C-terminal propeptide (PICP) were studied in 74 patients with various forms of non-lethal osteogenesis imperfecta and 27 unaffected family members. Using the standard deviation (SD) score, PICP concentrations were found to be > or = -1 SD in 16%, between -1 and -2 SD in 26% and < or = -2 SD in 58% of the patients with osteogenesis imperfecta compared to healthy controls. PICP values were lowest in osteogenesis imperfecta type I (-2.4 +/- 0.4 SD, n = 37) followed by type III (-1.9 +/- 0.5 SD, n = 13) and type IV ( 1.3 +/- 0.7 SD, n = 20). Four patients with osteogenesis imperfecta with an atypical clinical course had normal or even elevated levels which may indicate heterogeneity in the underlying primary defects. In osteogenesis imperfecta type I, PICP concentrations proved to be a helpful serum marker for pedigree screening. Osteocalcin was high in 25 of 28 patients with osteogenesis imperfecta in the first decade but only in 1 of 18 older patients. Insulin-like growth factor-I was within the normal range in 53 cases of osteogenesis imperfecta, decreased in 2 and elevated in 3 patients. We conclude that PICP concentration is a useful parameter in the clinical management of osteogenesis imperfecta, including the assessment of future therapeutic interventions. PMID- 8241675 TI - Severe hyperlipoproteinemia in congenital nephrotic syndrome of the Finnish type: effect of dialysis and kidney transplantation. AB - Two children with congenital nephrosis of the Finnish type were studied successively at the three stages of the disease: (A) nephrosis, (B) renal insufficiency/peritoneal dialysis and (C) post-transplantation; two additional patients were studied at two stages. Plasma lipoprotein profiles were determined by density gradient ultracentrifugation and lipids by enzymatic methods. Stage A was characterized by hyperchylomicronemia, low high density lipoprotein (HDL) cholesterol and the presence of dense low density lipoprotein (LDL) and HDL particles. Total cholesterol and triglycerides showed great daily variation (5-14 and 5-33 mmol/l, respectively). During stage B, hyperlipidemia weakened. Yet HDL concentration remained low and the concentration of intermediate density lipoproteins (IDL) increased. At stage C, hyperlipidemia had almost subsided, but the presence of IDL persisted. In conclusion, severe hyperlipoproteinemia of congenital nephrosis at the nephrotic stage is attenuated during renal insufficiency and dialysis, and essentially normalizes after kidney transplantation. Yet the presence of IDL implies an increased risk of atherosclerosis. PMID- 8241676 TI - Core-peripheral temperature gradient in children: does it reflect clinically important changes in circulatory haemodynamics? AB - Following open heart surgery, changes in core and peripheral skin temperature and changes in the core-peripheral temperature gradient were measured in 10 children. These were correlated with changes in cardiac index, systemic vascular resistance index, mean arterial pressure and urinary output. During the study intervals, which lasted 1 h each, no changes in medical management were made. Using Spearman's rank correlation, only a change in central venous pressure was found to correlate with a change in the core-peripheral temperature gradient. We conclude that a change in the core-peripheral temperature will give valuable information about the patient's intravascular volume. PMID- 8241677 TI - Alcohol intoxication in hospitalized young teenagers. AB - The scientific literature concerning alcohol intoxication is enormous. However, less is known of alcohol-induced disturbances in children and adolescents and most of those reports concern cases of hypoglycemia in children under five years of age. We studied the clinical status and chemistry, especially acid-base balance, in 36 young teenagers treated at hospital for alcohol intoxication. On physical examination 6 patients were somnolent, 18 were comatose and 12 were in deep coma. The impairment of consciousness was directly proportional to the blood ethanol concentration. Acidosis was a central finding, and it was caused by a combination of respiratory and metabolic factors (a high blood PCO2 and a low base excess; r = 0.97, p < 0.001); the finding of respiratory acidosis dominated. Base excess correlated negatively with beta-hydroxybutyrate and lactate, as expected. All the metabolic products measured--acetate, beta-hydroxybutyrate and lactate--were significantly elevated compared with the control patients. No hypoglycemia was found. Prior treatment with intravenous glucose decreased vomiting and normalized the serum lactate concentration and PO2. Hypokalemia was the most common abnormality in serum electrolytes. In four patients the rate of fall of blood ethanol concentration was 2.8-3.3 mmol/h (0.13-0.15 g/l-1 h-1) and the mean acetate concentration was 0.8 mmol/l (SE 0.3). Biochemical disturbances in young teenage alcohol intoxicants resemble those previously found in adults. The severe toxicity by ethanol, manifesting in coma, occurs in lower blood alcohol concentrations in children than in adults. PMID- 8241678 TI - Erythrocyte membrane fatty acid composition in children with atopic dermatitis compared to age-matched controls. PMID- 8241679 TI - Analgesic effect of lidocaine ointment on intact skin in neonates. PMID- 8241680 TI - Gentamicin-resistant Enterobacteriaceae in a neonatal intensive care unit. PMID- 8241681 TI - Kala-azar in a one-year-old Swedish child. Diagnostic difficulties because of active hemophagocytosis. AB - A one-year-old Swedish boy developed kala-azar six months after a holiday in Spain. Upon visiting the hospital after one week of illness he demonstrated clinical and laboratory findings of fever, splenomegaly and cytopenia. A fine needle aspiration biopsy of the spleen revealed hemophagocytosis and he had increased serum levels of the cytokines tumor necrosis factor-alpha and interferon-gamma. Initially, a diagnosis of hemophagocytic lymphohistiocytosis was made. Re-evaluation of the spleen smears and of the bone marrow aspiration revealed Leishmania parasites and subsequent therapy with sodium stibogluconate was successful. This patient illustrates the interesting similarities between these two disorders involving the mononuclear phagocyte system as well as the problems involved in differential diagnosis. This case also reminds us of the possibility of contracting visceral leishmaniasis in Mediterranean countries. PMID- 8241682 TI - Increase in T cells bearing the gamma/delta receptor associated with lymphoproliferative disease of granular lymphocytes in an infant with intractable diarrhea. AB - A two-year-old infant with intractable diarrhea and lymphoproliferative disease of granular lymphocytes attributed to a persistent cytomegalovirus infection showed an increase in cells bearing the gamma/delta T-cell receptor (TCR), which accounted for approximately 20% of total peripheral blood lymphocytes and 40% of CD3+ T cells. Of the gamma/delta TCR+ cells, two-thirds were double negative (CD4 /CD8-) and the other one-third CD8 positive. The majority of gamma/delta+ cells were delta TCS 1 positive. The predominance of delta TCS 1 positive cells was also confirmed on biopsy of lymphoid tissues from the colon. After improvement of watery diarrhea and malnutrition following three-month hyperalimentation, the number of gamma/delta TCR+ cells decreased. The patient subsequently died of pneumonia at the age of 2 years and 11 months. A possible site-specific role for the gamma/delta TCR+ cells, particularly delta TCS 1+ cells, in the human intestine is discussed. PMID- 8241683 TI - Observations on very high-dose steroid therapy in nephrotic syndrome. PMID- 8241684 TI - Clinical continuing education for school nurses. AB - Because of the increased presence of medically complex students in schools, school nurses need to maintain a high level of clinical expertise and continuing education must now include the technical and clinical aspects of care. The authors describe a unique continuing education program designed to update the theoretical knowledge and clinical skills of school nurses caring for students with respiratory disabilities. This report focuses on the development, implementation, and evaluation of a continuing education program with a specific clinical component. PMID- 8241685 TI - Adolescent substance abuse: psychosocial factors. AB - The prevention and management of adolescent substance abuse requires an understanding of the psychosocial context in which such a problem develops. This article reviews literature from the health and behavioral sciences and provides information about the prevalence of substance abuse, family and other psychosocial factors associated with substance abuse, and signs and symptoms of various kinds of substance abuse. Implications for school nursing practice are discussed and suggestions for implementation made. PMID- 8241686 TI - An overview for school nursing professionals: child sexual abuse. PMID- 8241687 TI - A position statement of the National Association of State School Nurse Consultants. Medicaid reimbursement for school nursing services. August 1993. AB - The National Association of State School Nurse Consultants (NASSNC) believes that school nursing services should be provided to all students and that Medicaid funds should be incorporated as a funding source for school nursing services provided to eligible children. The list of nursing services and procedures in this document, therefore, should be reimbursable by Medicaid to school districts. These services are outlined in categories of 1) case finding, 2) nursing care procedures, 3) care coordination, 4) patient/student counseling/instruction, and 5) emergency care. We adhere to this philosophy and promote incorporation of these services as Medicaid-reimbursable services provided by qualified professional school nurses. PMID- 8241688 TI - Challenge for the 90's. PMID- 8241690 TI - To compute or not to compute. PMID- 8241689 TI - Nursing practice management: the numbers of children classified as homeless are growing in our schools. PMID- 8241691 TI - Problem solving styles of school nurses. AB - School nurses are challenged on a daily basis to solve complex problems and generate creative solutions to meet the health needs of children. This study focused on how school nurses go about solving problems in the delivery of school health services. The results showed that school nurses' problem-solving style allows them to act as "bridgers" within the educational environment and to promote collaboration among many types of school personnel. PMID- 8241692 TI - Coenzyme Q10 protects ischemic myocardium in an open-chest swine model. AB - Myocardial stunning, defined as a reversible decrease in contractility after ischemia and reperfusion, may be a manifestation of reperfusion injury caused by free oxygen radical damage. The aim of this study was to test the hypothesis that pretreatment with coenzyme Q10 (ubiquinone), believed to act as a free radical scavenger, reduces myocardial stunning in a porcine model. Twelve swine were randomized to receive either oral supplementation with coenzyme Q10 or placebo for 20 days. A normothermic open-chest model was used with short occlusion (8 min) of the distal left descending coronary artery followed by reperfusion. Regional contractile function was measured with epicardial Doppler crystals in ischemic and nonischemic segments by measuring thickening fraction of the left ventricular wall during systole. Stunning time was defined as the elapsed time of reduced contractility until return to baseline. Coenzyme Q10 concentrations were measured in blood and homogenized myocardial tissue by high performance liquid chromatography. Plasma levels of reduced coenzyme Q10 (ubiquinol) were higher in swine pretreated with the experimental medication as compared to placebo (mean 0.45 mg/l versus 0.11 mg/l, respectively). Myocardial tissue concentrations, however, did not show any changes (mean 0.79 micrograms/mg dry weight versus 0.74 micrograms/mg). Stunning time was significantly reduced in coenzyme Q10 pretreated animals (13.7 +/- 7.7 min versus 32.8 +/- 3.1 min, P < 0.01). In conclusion, chronic pretreatment with coenzyme Q10 protects ischemic myocardium in an open-chest swine model. The beneficial effect of coenzyme Q10 on myocardial stunning may be due to protection from free radical mediated reperfusion injury. This protective effect seems to be generated by a humoral rather than intracellular mechanism. PMID- 8241693 TI - Coenzyme Q10 and coronary artery disease. AB - It has been postulated that oxidatively modified low-density lipoprotein (LDL) contributes to the genesis of atherosclerosis. Ubiquinone has been suggested to be an important physiological lipid-soluble antioxidant and is found in LDL fractions in the blood. We measured plasma level of ubiquinone using high performance liquid chromatography and plasma levels of total cholesterol, high density lipoprotein (HDL) cholesterol, and triglycerides in 245 normal subjects (186 males, 59 females) and in 104 patients (55 males, 49 females) who had coronary artery disease not receiving pravastatin and 29 patients (12 males, 17 females) receiving pravastatin. In the normal subjects, the plasma ubiquinone levels did not vary with age. In the patient groups, the plasma total cholesterol and LDL levels were higher and the plasma ubiquinone level lower than in the normal subject group. The LDL/ubiquinone ratio was higher in the patient groups. We found that ubiquinone level, either alone or when expressed in relation to LDL levels, was significantly lower in the patient groups compared with the normal subject group. The 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitor is thought to prevent atherosclerosis, however, it also inhibits ubiquinone production. The present study revealed that HMG CoA reductase inhibitor decreased plasma cholesterol level, and that it did not improve either the ubiquinone level or the LDL/ubiquinone ratio. From these results, the LDL/ubiquinone ratio is likely to be a risk factor for atherogenesis, and administration of ubiquinone to patients at risk might be needed. PMID- 8241694 TI - Perspectives on therapy of cardiovascular diseases with coenzyme Q10 (ubiquinone). AB - A defective myocardial energy supply--due to lack of substrates and/or essential cofactors and a poor utilization efficiency of oxygen--may be a common final pathway in the progression of myocardial diseases of various etiologies. The vitamin-like essential substance coenzyme Q10, or ubiquinone, is a natural antioxidant and has a key role in oxidative phosphorylation. A biochemical rationale for using coenzyme Q10 as a therapy in heart disease was established years ago by Folkers and associates; however, this has been further strengthened by investigations of viable myocardial tissue from the author's series of 45 patients with various cardiomyopathies. Myocardial tissue levels of coenzyme Q10 determined by high-performance lipid chromatography were found to be significantly lower in patients with more advanced heart failure compared with those in the milder stages of heart failure. Furthermore, the myocardial tissue coenzyme Q10 deficiency might be restored significantly by oral supplementation in selected cases. In the author's open clinical protocol study with coenzyme Q10 therapy (100 mg daily) nearly two-thirds of patients revealed clinical improvement, most pronounced in those with dilated cardiomyopathy. Double-blind placebo-controlled trials have definitely confirmed that coenzyme Q10 has a place as adjunctive treatment in heart failure with beneficial effects on the clinical outcome, the patients' physical activity, and their quality of life. The positive results have been above and beyond the clinical status obtained from treatment with traditional principles--including angiotensin-converting enzyme inhibitors. PMID- 8241695 TI - Role of metabolic therapy in cardiovascular disease. AB - The pathophysiological basis for the use of metabolic therapy in the treatment of heart failure is analyzed. Bioenergetical processes related to ATP bioavailability play a central role in regulating myocardial contractility at rest and on effort. Furthermore, a significant correlation has been demonstrated in diseased heart between ATP content, revealed at endomyocardial biopsy, and systolic and diastolic left ventricular indexes evaluated with invasive and noninvasive methods. Several international investigations demonstrate the beneficial effects of ubiquinone (coenzyme Q10) in the treatment of heart failure. Here the results of a study are reported that was conducted on patients with heart failure treated with ubiquinone. After 7 months of oral drug administration (100 mg/day), a significant improvement was observed in echocardiographic indexes of systolic function, cardiothoracic ratio, and clinical signs and symptoms of congestive heart failure. In conclusion, the introduction of metabolic drugs, such as ubiquinone, in the treatment of heart failure opens new horizons in the therapeutic approach to an ailment that entails substantial human and social costs. PMID- 8241696 TI - Italian multicenter study on the efficacy and safety of coenzyme Q10 as adjuvant therapy in heart failure. AB - This multicenter study evaluated the efficacy and tolerability of coenzyme Q10 in 1715 outpatients with chronic heart failure (New York Heart Association classes II and III), stabilized with standard therapy for 3 months. The patients were treated with coenzyme Q10 at a daily dose of 50 mg for 4 weeks, in addition to receiving conventional therapy. The efficacy of coenzyme Q10 was assessed by an open study that evaluated the improvement in clinical signs and symptoms of heart failure. After the baseline evaluation the subjects were seen on days 15 and 30. The intensity of signs and symptoms was assessed by a semiquantitative 4-point scale. Our results demonstrate that the administration of coenzyme Q10 in association with standard therapy improves dyspnea at rest, exertional dyspnea, palpitations, cyanosis, hepatomegaly, pulmonary rales, ankle edema, heart rate, and systolic and diastolic blood pressure in patients with stabilized heart failure. The rate of improvement and the low number of side effects in this large group of patients demonstrate that despite some methodological limitations in the study design and the short period of treatment (4 weeks) coenzyme Q10 given at a daily dose of 50 mg led to an improvement in the signs and symptoms of heart failure and in the quality of life. PMID- 8241697 TI - Effect of coenzyme Q10 therapy in patients with congestive heart failure: a long term multicenter randomized study. AB - The improved cardiac function in patients with congestive heart failure treated with coenzyme Q10 supports the hypothesis that this condition is characterized by mitochondrial dysfunction and energy starvation, so that it may be ameliorated by coenzyme Q10 supplementation. However, the main clinical problems in patients with congestive heart failure are the frequent need of hospitalization and the high incidence of life-threatening arrhythmias, pulmonary edema, and other serious complications. Thus, we studied the influence of coenzyme Q10 long-term treatment on these events in patients with chronic congestive heart failure (New York Heart Association functional class III and IV) receiving conventional treatment for heart failure. They were randomly assigned to receive either placebo (n = 322, mean age 67 years, range 30-88 years) or coenzyme Q10 (n = 319, mean age 67 years, range 26-89 years) at the dosage of 2 mg/kg per day in a 1 year double-blind trial. The number of patients who required hospitalization for worsening heart failure was smaller in the coenzyme Q10 treated group (n = 73) than in the control group (n = 118, P < 0.001). Similarly, the episodes of pulmonary edema or cardiac asthma were reduced in the control group (20 versus 51 and 97 versus 198, respectively; both P < 0.001) as compared to the placebo group. Our results demonstrate that the addition of coenzyme Q10 to conventional therapy significantly reduces hospitalization for worsening of heart failure and the incidence of serious complications in patients with chronic congestive heart failure. PMID- 8241698 TI - Statistical data support prediction of death within 6 months on low levels of coenzyme Q10 and other entities. AB - Ninety-four consecutive hospital patients aged over 50 years were included in a cross-sectional study. Serum samples were analyzed for coenzyme Q10, alpha tocopherol, and free cholesterol levels. Patients who died within a follow-up period of 6 months or had congestive heart failure or severe myalgia, and/or received cytostatic or lipid-lowering drug therapy showed significantly lower free cholesterol-related coenzyme Q10 values. Prospective controlled clinical trials will determine whether coenzyme Q10 has a potential to protect patients from such complications and become a useful therapy. PMID- 8241699 TI - Isolated diastolic dysfunction of the myocardium and its response to CoQ10 treatment. AB - Symptoms of fatigue and activity impairment, atypical precordial pain, and cardiac arrhythmia frequently precede by years the development of congestive heart failure. Of 115 patients with these symptoms, 60 were diagnosed as having hypertensive cardiovascular disease, 27 mitral valve prolapse syndrome, and 28 chronic fatigue syndrome. These symptoms are common with diastolic dysfunction, and diastolic function is energy dependent. All patients had blood pressure, clinical status, coenzyme Q10 (CoQ10) blood levels and echocardiographic measurement of diastolic function, systolic function, and myocardial thickness recorded before and after CoQ10 replacement. At control, 63 patients were functional class III and 54 class II; all showed diastolic dysfunction; the mean CoQ10 blood level was 0.855 micrograms/ml; 65%, 15%, and 7% showed significant myocardial hypertrophy, and 87%, 30%, and 11% had elevated blood pressure readings in hypertensive disease, mitral valve prolapse and chronic fatigue syndrome respectively. Except for higher blood pressure levels and more myocardial thickening in the hypertensive patients, there was little difference between the three groups. CoQ10 administration resulted in improvement in all; reduction in high blood pressure in 80%, and improvement in diastolic function in all patients with follow-up echocardiograms to date; a reduction in myocardial thickness in 53% of hypertensives and 36% of the combined prolapse and fatigue syndrome groups; and a reduced fractional shortening in those high at control and an increase in those initially low.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241700 TI - Italian multicenter study on the safety and efficacy of coenzyme Q10 as adjunctive therapy in heart failure (interim analysis). The CoQ10 Drug Surveillance Investigators. AB - Digitalis, diuretics, and vasodilators are considered standard therapy for patients with congestive heart failure, for which treatment is tailored according to the severity of the syndrome and the patient profile. Apart from the clinical seriousness, heart failure is always characterized by an energy depletion status, as indicated by low intramyocardial ATP and coenzyme Q10 levels. We investigated safety and clinical efficacy of coenzyme Q10 (CoQ10) adjunctive treatment in congestive heart failure, which had been diagnosed at least 6 months previously and treated with standard therapy. A total of 2500 patients in NYHA classes II and III were enrolled in this open noncomparative 3-month postmarketing drug surveillance study in 173 Italian centers. The daily dose of CoQ10 was 50-150 mg orally, with the majority of patients (78%) receiving 100 mg/day. Clinical and laboratory parameters were evaluated at the entry into the study and on day 90; the assessment of clinical signs and symptoms was made using from two- to seven point scales. Preliminary results on 1113 patients (mean age 69.5 years) show a low incidence of side effects: 10 adverse reactions were reported in 8 (0.8%) patients, of which only 5 reactions were considered as correlated to the test treatment. After 3 months of test treatment the proportions of patients with improvement in clinical signs and symptoms were as follows: cyanosis 81%, edema 76.9%, pulmonary rales 78.4%, enlargement of the liver area 49.3%, jugular reflux 81.5%, dyspnea 54.2%, palpitations 75.7%, sweating 82.4%, arrhythmia 62%, insomnia 60.2%, vertigo 73%, and nocturia 50.7%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241701 TI - Recovery of the Frank-Starling mechanism by coenzyme Q10 in patients with load induced contractility depression. AB - Load-induced contractility depression, in which supernormal left ventricular ejection fraction and contractility at rest decrease by added afterload, is most often found in children with mitral valve prolapse who have symptoms. Patients have high ventricular end-diastolic pressure at rest, which is further increased by afterload challenge. The Frank-Starling mechanism may be maximally mobilized with high preload even at rest to compensate for the intrinsically depressed inotropic state. Therefore, preload reserve may be easily exhausted due to afterload addition. We aimed to determine left ventricular end-diastolic fiber length, stroke work, and contractility before and during handgrip by echocardiograms to obtain evidence for the Frank-Starling mechanism in patients and controls, including patients treated with coenzyme Q10. The subjects were divided into four groups, each consisting of 30 children aged 6-16 years: group 1, normals; group 2, patients; group 3, the same patients as in group 2 after coenzyme Q10 therapy; and group 4, patients with asymptomatic mitral valve prolapse. Baseline values and percentage increases in systolic blood pressure, heart rate, and left ventricular wall stress showed no differences among the groups. Only in group 2 were the percentage increase in ejection fraction, fiber shortening velocity, contractility, and end-diastolic dimension strongly negative, despite supernormal baseline levels. In other groups, these were significantly positive, without intergroup differences. We conclude that in the heart with load-induced contractility depression, the Frank-Starling mechanism deviates from normal. The normal Frank-Starling mechanism was recovered due to coenzyme Q10, which may improve disturbed bioenergetic function at the molecular level. PMID- 8241702 TI - Myocardial preservation by therapy with coenzyme Q10 during heart surgery. AB - Coenzyme Q10 (CoQ10) is a natural and essential cofactor in the heart. It is the primary redox coupler in the respiratory chain, a potent free radical scavenger, and a superoxide inhibitor. In this study the myocardial protective effects of CoQ10 were determined in high-risk (n = 10) patients during heart surgery compared to that found in placebo controls (n = 10). In both groups, there was a blood CoQ10 deficiency (< 0.6 microgram/ml), low cardiac index (CI < 2.4 l/m2 per minute), and low left ventricular ejection fraction (LVEF < 35%) before treatment. CoQ10 (100 mg per day) was given orally for 14 days before and 30 days after surgery. Presurgical CoQ10 treatment significantly (P < 0.01) improved blood and myocardial CoQ10 and myocardial ATP compared to that found in the control group. Cardiac functions (CI and LVEF) were improved but not significantly. After cardiac cooling, rewarming, and reperfusion; blood and tissue CoQ10 and tissue ATP levels were maintained in the normal ranges in the CoQ10 patients. Cardiac pumping (CI) and LVEF were significantly (P < 0.01) improved. The recovery course was short (3-5 days) and uncomplicated. In the control group blood and tissue CoQ10, tissue ATP levels, and cardiac functions were depressed after surgery. The recovery course was long (15-30 days) and complicated. Positive relationships between blood and myocardial CoQ10, myocardial ATP, cardiac function, and the postoperative recovery time and course found in both study groups show the therapeutic benefits of CoQ10 in preserving the myocardium during heart surgery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241703 TI - Effects of coenzyme Q10 administration on pulmonary function and exercise performance in patients with chronic lung diseases. AB - Serum coenzyme Q10 (CoQ10) levels were measured at rest and during incremental exercise in 21 patients with chronic obstructive pulmonary disease (COPD) and 9 patients with idiopathic pulmonary fibrosis (IPF). The mean serum CoQ10 levels at rest in patients with COPD and IPF were 0.56 +/- 0.20 and 0.45 +/- 0.16 microgram/ml, respectively. In both groups these levels were decreased compared with those of healthy subjects. In the patients with COPD, CoQ10 levels were significantly correlated with body weight, however, there was no correlation between CoQ10 levels and ventilatory function, PaO2, VO2/kg at rest, or maximal VO2. In eight of nine patients whose PaO2 at rest was lower than 75 torr, serum CoQ10 levels were lower than 0.5 microgram/ml. We studied the effects of the oral administration of CoQ10 at 90 mg/day for 8 weeks on pulmonary function and exercise performance in eight patients with COPD. Serum CoQ10 levels were significantly elevated in association with an improvement in hypoxemia at rest, whereas pulmonary function was unaltered. Oxygen consumption during exercise was not changed, whereas PaO2 was significantly improved, and heart rate was significantly decreased compared with the results obtained at an identical workload at baseline. Furthermore, lactate production was suppressed during the anaerobic exercise stage after CoQ10 administration, and exercise performance tended to increase. These data suggested that CoQ10 has favorable effects on muscular energy metabolism in patients with chronic lung diseases who have hypoxemia at rest and/or during exercise. PMID- 8241704 TI - A short historical survey of heart failure. PMID- 8241705 TI - Heart failure is a dominant deficiency of coenzyme Q10 and challenges for future clinical research on CoQ10. PMID- 8241706 TI - The essential functions of coenzyme Q. AB - The essential role of coenzyme Q in biological energy transduction is well established. Coenzyme Q is a unique carrier for two-electron transfer within the lipid phase of the mitochondrial membrane. The function is essential for proton based energy coupling. The sites of entry and exit of electrons into the quinone are at specific quinone-binding sites which are constructed to allow only two electron transfer and thus prevent damaging free radical formation by direct reaction of oxygen with the semiquinone. Failure of proper function with diminished energy supply can be related to insufficient quinone, modification of lipid fluidity, or lipid protein interaction and damage or poisoning in binding sites. Supplementation with coenzyme Q can act by reversal of deficiency or decreased mobility, or by overcoming binding site modification. Coenzyme Q has also been shown to increase antioxidant protection in membranes. New sites for coenzyme Q function in Golgi and plasma membrane show evidence for a role in growth control and secretion-related membrane flow. PMID- 8241707 TI - Ubiquinol: an endogenous antioxidant in aerobic organisms. AB - Ubiquinone (coenzyme Q), in addition to its function as an electron and proton carrier in mitochondrial and bacterial electron transport linked to ATP synthesis, acts in its reduced form (ubiquinol) as an antioxidant, preventing the initiation and/or propagation of lipid peroxidation in biological membranes and in serum low-density lipoprotein. The antioxidant activity of ubiquinol is independent of the effect of vitamin E, which acts as a chain-breaking antioxidant inhibiting the propagation of lipid peroxidation. In addition, ubiquinol can efficiently sustain the effect of vitamin E by regenerating the vitamin from the tocopheroxyl radical, which otherwise must rely on water-soluble agents such as ascorbate (vitamin C). Ubiquinol is the only known lipid-soluble antioxidant that animal cells can synthesize de novo, and for which there exist enzymic mechanisms that can regenerate the antioxidant from its oxidized form resulting from its inhibitory effect of lipid peroxidation. These features, together with its high degree of hydrophobicity and its widespread occurrence in biological membranes and in low-density lipoprotein, suggest an important role of ubiquinol in cellular defense against oxidative damage. Degenerative diseases and aging may be manifestations of a decreased capacity to maintain adequate ubiquinol levels. PMID- 8241708 TI - The function of coenzyme Q in mitochondria. AB - We have accumulated evidence that coenzyme Q (CoQ) concentration in the mitochondrial membrane is not saturating for NADH oxidation but is saturating for succinate and glycerol-3-phosphate oxidation. As a result of its kinetic properties CoQ concentration changes must yield changes in respiration rates. This provides a rationale for the reported therapeutic effects of CoQ under conditions when its concentration is decreased, as has been reported in tissues from aged rats; we have failed, however, to detect any specific CoQ decrease in mitochondria from several tissues of aged rats. We can, however, predict from the kinetic bases that CoQ would ameliorate respiration rate also under conditions in which a defect is present in regions not involving the quinone. CoQ incorporation in perfused liver is attempted in order to find experimental systems for investigating its protecting effect. Liposomal CoQ10 perfused in rat livers (where CoQ9 is the main homolog) is incorporated mainly in lysosomes, and its increase in the crude mitochondrial fraction could be mainly ascribed to residual lysosomal contamination. Nevertheless, perfusion with exogenous CoQ10 maintains higher levels of endogenous CoQ9, and higher glutamate oxidation than in controls. In the same system, an oxidative stress by doxorubicin induces mitochondrial changes, including a decrease in endogenous CoQ9 and in respiratory activities. These changes are prevented by concomitant perfusion of liposomal CoQ10. PMID- 8241709 TI - Cardiostimulatory action of coenzyme Q homologues on cultured myocardial cells and their biochemical mechanisms. AB - The effect of coenzyme Q (CoQ) homologues on the beating of myocardial cells was investigated in cultured cell sheets from mouse fetuses and quail embryos. Myocardial cell sheets grown in Eagle's minimum essential medium with fetal bovine serum showed very weak and irregular beating when this serum was removed from the medium. However, the depressed beating rate and amplitude recovered almost completely within a few minutes by adding CoQ10 to the medium, and the effect of CoQ10 continued over 1 h. CoQ9 showed a cardiostimulatory effect similar to that of CoQ10, but CoQ8 and CoQ7 showed almost no effect. Short homologues (less than CoQ4) inhibited the beating of cell sheets. The cardiostimulatory effect of CoQ10 was not blocked by atenolol, a selective beta blocker. In addition, CoQ10 stimulated the formation of ATP, not cAMP. CoQ0 and CoQ3 inhibited beating rates by inhibiting ATP formation. In conclusion, only native CoQ homologues having a nona- or decaprenyl group showed a cardiostimulatory effect on cultured myocardial cells, probably by stimulating mitochondrial ATP formation. PMID- 8241710 TI - Heart muscle ubiquinone and plasma antioxidants following cardiac transplantation. AB - Nine patients who underwent heart transplantation (one female; average age 48 +/- 11, range 19-58 years) were followed in respect to contents of right-sided heart septum, blood and plasma ubiquinone (UQ), plasma alpha-tocopherol (alpha T), and plasma free cholesterol (FC). In contrast to healthy persons, substantial inter- and intraindividual variations were observed; individually low values were seen with rejection. Heart muscle UQ in well-treated patients averaged 0.33 +/- 0.08, range 0.06-0.58 micrograms mg-1 (0.38 +/- 0.09 mumol g-1 dry weight) and was not different from healthy individuals. Plasma UQ, alpha T; and FC averaged 0.63 +/- 0.33 micrograms ml-1 (P < 0.05 versus sedentary controls), 8.1 +/- 4.0 micrograms ml-1 (P < 0.01), and 0.52 +/- 0.23 mg ml-1 (P < 0.05). Corresponding molar values were 0.73 +/- 0.37 (UQ), 2.0 +/- 1.1 mumol l-1 (alpha T), and 1.42 +/- 0.54 mmol l-1 (FC). Blood and plasma UQ values were identical. A saturation like relationship was found between heart and blood UQ:blood contents below 0.7 micrograms ml-1 (0.8 mumol l-1) corresponded to markedly lowered heart contents. In four patients in whom blood samples were taken close to a fatal complication it averaged 0.42 micrograms ml-1 (0.49 mumol l-1, P < 0.01). When low heart muscle and blood ubiquinone were present, other variables such as left ventricle cardiac output or cycle ergometer performance was markedly impaired. Plasma UQ and alpha T covaried with a marker of the lipoidal deposit volume, plasma FC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241711 TI - Ubiquinone and alpha-tocopherol in plasma; means of translocation or depot. AB - Ubiquinone (UQ) and alpha-tocopherol (AT) are two highly lipophilic antioxidants which can be dissolved only in lipid layers or attached to protein structures. Analyses of both UQ and AT in whole blood and plasma demonstrate identical values, which excludes any significant allocation to blood cells. The lipoidic plasma structures constitute the plasma lipoprotein fractions of high (HDL), low (LDL), and very low (VLDL) density in addition to chylomicrons. This means by definition that blood and plasma UQ and AT values are limited if not related to the lipoidic deposit volume. UQ and AT increase linearly with free cholesterol (FC). FC has therefore been suggested to be a good marker for the deposit volume. The ratios UQ and AT over FC--normalized UQ (N-UQ) and normalized AT (N-AT)--have been computed for inter- and intraindividual comparisons. With a plasma UQ content of 1 microgram/ml (approximately 1 mumol/l) and a plasma volume of 41, UQ makes up about 15% of the total heart content or under 1% of UQ in skeletal muscle. The corresponding value for the total extracellular UQ content is less than 2%. This means that extracellular UQ has no or a very minor role as a UQ depot. The same is true for AT. However, for transportation and allocation determinations of N-UQ and N-AT are relevant. Assuming only a lipoprotein-related transportation, healthy persons have saturated plasma UQ and AT values in only 25% and 10% of the population, respectively. All patient categories studied have been found nonsaturated. VLDL plus LDL constitute some 90% of the UQ deposit volume.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241712 TI - Antioxidant effect of coenzyme Q on hydrogen peroxide-activated myoglobin. AB - In recent years increased attention has been focused on the reduced forms of coenzyme Q as antioxidant compounds inhibiting lipid peroxidation in model systems and in biological membranes, but in spite of extensive experimental evidences the molecular mechanisms responsible for the antioxidant activity of ubiquinones are still debated. Ferrylmyoglobin and/or its free radical form are regarded as powerful oxidizing agents capable of promoting oxidation of essential cellular constituents, particularly cell membranes. Therefore, we investigated the effects of ubiquinol on the formation and survival of ferryl species of myoglobin and on metmyoglobin itself. The addition of a threefold molar excess of hydrogen peroxide to a solution of metmyoglobin induces the rapid formation of a compound with the spectral characteristics of ferrylmyoglobin. The reaction is complete within 4 min, producing up to 76% of ferrylmyoglobin, which remains stable for at least 30 min. The addition of ubiquinol-1 to the same solution provokes a rapid and progressive reduction of ferrylmyoglobin to metmyoglobin and oxymyoglobin. Ubiquinol-1, furthermore, is also capable of protecting metmyoglobin against oxidation when added in the solution before hydrogen peroxide. Ubiquinol-1, indeed, is effective at both limiting the maximal ferrylmyoglobin level attained (59% inhibition) and accomplishing complete removal of the ferryl form (in about 15 min). The results demonstrate that ubiquinol is capable of reducing both ferrylmyoglobin and metmyoglobin to oxymyoglobin, providing a novel antioxidant mechanism for coenzyme Q. PMID- 8241714 TI - MIR '92--a new era in space exploration. PMID- 8241713 TI - Effects of inhibitors of hydroxymethylglutaryl coenzyme A reductase on coenzyme Q and dolichol biosynthesis. AB - Inhibitors of hydroxymethylglutaryl coenzyme A reductase are used clinically to decrease blood levels of low-density lipoprotein cholesterol in hypercholesterolemic patients. However, little is known about the possible effects of these inhibitors on dolichol and cholesterol synthesis. Oral administration of mevinolin to rats was found here to decrease dolichol, dolichyl P and coenzyme Q levels in the heart and skeletal muscle and to increase the hepatic dolichol level while decreasing the coenzyme Q content in this same organ. The amounts of dolichyl-P decreased in heart and muscle and increased in brain. Intraperitoneal administration also affected the levels of these lipids. The concentrations of blood lipids were not modified in the same manner as tissue lipids. Analysis of individual enzyme activities and of incorporation of [3H]acetate into various lipids of liver and brain slices demonstrated that both up- and down-regulation of different proteins occur in various tissues, resulting in modifications in lipid synthesis. Hypercholesterolemic patients were found to have high blood coenzyme Q levels, which are decreased upon pravastatin treatment, although they are still above control values. It appears that these HMG-coenzyme A reductase inhibitors do not selectively lower cholesterol levels, but that they also modify the dolichol and coenzyme Q content and synthesis both in the liver and various other tissues. PMID- 8241715 TI - MIR '92: a joint Russian-German space mission. PMID- 8241716 TI - Reduced natriuresis during weightlessness. AB - The kidney response to weightlessness was measured in one volunteer during a 1 week space mission. Shortly after entering microgravity and later during the mission, consecutive urine sampling periods were monitored, covering in total about 50% of the inflight time. Preflight references were a sequence of ground based experiments, which evaluated body fluid metabolism with different degrees of standardization. Additional variables, such as circadian rhythms and cortisol associated stress, were also monitored. In contrast to current hypotheses, the volunteer showed a pronounced reduction in natriuresis and diuresis during the entire space flight, despite a considerable weight loss. For the first time, the urinary excretion of the renal natriuretic peptide urodilatin was also measured. Both, during the preflight experiments and during weightlessness, close correlations between urodilatin excretion and sodium excretion were observed. However, the correlation between natriuresis and urodilatin excretion was considerably altered during weightlessness. We conclude that the loss of body weight during space flight is not related to an increased renal fluid loss and that urodilatin might counteract the decrease in renal excretion observed in weightlessness. PMID- 8241717 TI - Fluid shifts into and out of superficial tissues under microgravity and terrestrial conditions. AB - The microgravity environment can be expected in man to induce a swelling of facial tissues and a shrinking of the tissues in the lower limbs together with a loss in body weight. To quantitate fluid shifts into and out of superficial tissues an ultrasound A-mode method was used in one cosmonaut during a 7-day spaceflight. Measurements were taken from frontal and tibia tissues, where the underlying bone provides a good backwall echo. During the spaceflight the cosmonaut showed a swelling of facial tissues during the first 3 days. At the same time the superficial tissues of the caudal areas shrank by 20%. In space he lost 7.7% of body weight. After the spaceflight the superficial tissues were dried out but regained their water content within the next 4 days even before body weight returned to control level. Per kilogram of body weight an increase of 400 cm3 entered the superficial tissue layers of the body. It is concluded that water loss as well as wasting of tissues contribute to the loss in body weight during space flight. PMID- 8241718 TI - Body fluid distribution in man in space and effect of lower body negative pressure treatment. AB - The lack of hydrostatic forces in space eventually produces a fluid deficit within the circulatory system. This deficit may alter the circulatory regulation patterns. The aim of the present study was to determine how much of this fluid deficit is attributable to interstitial fluid losses and to determine the effects of lower body negative pressure (LBNP) treatment on fluid distribution. The body fluid distribution of one subject was assessed before, during, and after weightlessness using two electrical impedance methods: (a) standard quadripole impedance for the segments of upper torso, lower torso, thigh, and calf and (b) an electrical impedance tomography technique (applied potential tomography) for a thigh cross-section. To assess the content of interstitial free fluid a thigh cuff overlying the electrodes for applied potential tomography was inflated to suprasystolic values to ascertain how much fluid can be squeezed out of blood vessels and tissue of skin and muscle. After the first thigh cuff maneuver (CUFF I) the subject performed a cardiovascular stress test with LBNP to mimic the gravity-induced blood shift to the lower part of the body. Then the compression maneuver was repeated (CUFF II). (a) This experimental sequence demonstrated a reduction in interstitial fluid in weightlessness of roughly 40% at the thigh. (b) The CUFF I and LBNP experiment demonstrated a reduced ability to cope with blood pooling in microgravity. (c) The CUFF II experiment suggests that LBNP in microgravity can refill the interstitial spaces and counteract the associated cardiovascular deterioration. The impedance measurements provided estimates of the contribution of different body sections to the observed body weight loss of more than 6 kg. The chest contributed nothing of significance, the lower torso more than 0.5 l, and both calves roughly 1.5 l. The thigh segments of both legs contributed between 1.5 l and 2.0 l with an interstitial free fluid reduction in muscle and skin by 40%. PMID- 8241719 TI - Self-tonometry under microgravity conditions. AB - During a space mission flown in microgravity early changes in intraocular pressure due to the fluid shift effect in microgravity were analyzed with an automatic self-tonometer. In total, 13 pressure readings per eye were obtained during the mission. The first pressure readings 16 min after reaching microgravity revealed a 92% rise in intraocular pressure compared to the daytime correlated baseline data on earth followed by an adaptation phase lasting for several hours. Diurnal pressure profiles during the mission showed no relevant difference in altitude and amplitude. PMID- 8241720 TI - Effects of microgravity on interstitial muscle receptors affecting heart rate and blood pressure during static exercise. AB - Afferent nerve fibers from receptors situated in the interstitium of skeletal muscles can induce cardiovascular reflexes. It has been shown that these interstitial muscle receptors are also sensitive to the local state of hydration: increased heart rates and blood pressure values were seen during dynamic and static exercise after local dehydration on earth. Since weightlessness leads to a persisting fluid loss in the lower part of the body, we hypothesized that leg exercise in space would augment heart rate and blood pressure responses to a similar extent as during local, interstitial dehydration on earth. Initial measurements during weightlessness were obtained in one subject after 6 days of space flight. Heart rate and blood pressure responses to light static foot plantar flexion (18% of maximal voluntary contraction) were recorded in two sessions. To eliminate the influence of muscle perfusion, exercise was performed during a period of arterial occlusion obtained by means of pneumatic cuffs at mid thigh level. Identical protocols were used in the pre- and postflight controls, which were performed both in the sitting posture and in a -90 degrees tilted sitting posture assumed 30-40 min before arterial occlusion. During weightlessness the exercise responses of heart rate and systolic and diastolic blood pressure closely followed the tracings obtained with the tilted sitting posture on ground. The response amplitudes in these states of reduced lower limb volumes (about 20/min and 20 mmHg, respectively) exceeded the responses in the supine position by a factor of at least 2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241721 TI - Dosimetry in the space radiation field. AB - The results of dosimetric measurements are presented which were performed as part of a German experiment package flown onboard the Russian space station MIR. These results are compared to those of previous missions: the first United States Spacelab mission and the first German Spacelab mission. Detector packages consisting of plastic nuclear track detectors, nuclear emulsions, and thermoluminescence dosimeters were exposed in different sections of the Russian space station. The equivalent dose for the astronauts was calculated from the measurements to be 3.9 mSv. Before and after the flight venous blood was taken from the astronauts. Chromosomal aberrations in peripheral lymphocytes were analyzed. It was found that the radiation exposure during the spaceflight leads to an elevation of dicentric chromosomes, indicating a radiation burden of the astronauts. PMID- 8241722 TI - Sleep and circadian rhythm during a short space mission. AB - An experiment was conducted to assess sleep and circadian regulation in an orbiting spacecraft. In orbit the weakened influence of 24-h zeitgebers could result in delayed circadian phases with the possibility of a transition to free running circadian rhythms. This and the specific stressors of a space mission may lead to changes in ultradian sleep regulation and in reduced sleep quantity and quality. During the mission sleep was recorded polygraphically on tape, as was body temperature. Daytime alertness was rated subjectively by a mood questionnaire. For comparison the same parameters were measured during a baseline period preceding the space mission. The circadian rhythms of body temperature and alertness were found to be delayed in space compared to baseline. This may mark a phase shift or the transition to a circadian state of free-run. Sleep was shorter and more disturbed. The structure of sleep was significantly altered. In space REM latency was shorter, there was less REM sleep in the second non-REM/REM cycle, and slow-wave sleep was redistributed from the first to the second cycle. The self-assessed mood resembled sleep disturbances and adaptation to the space environment. Reduced sleep quality and quantity are likely to result in fatigue and lower daytime performance. Countermeasures should be adopted to improve sleep of astronauts. PMID- 8241723 TI - Behavioral aspects of human adaptation to space: analyses of cognitive and psychomotor performance in space during an 8-day space mission. AB - Living in the space environment and exposure to microgravity induce a number of effects that may interfere with human cognitive and psychomotor performance. However, up to now very few attempts have been made to monitor possible impairments of human performance during space missions. In the present single case study several cognitive and psychomotor functions were monitored during an 8 day space mission to an orbital station using a computerized performance monitoring device. This device included four different tasks, which were selected from a battery of Standardized Tests for Research with Environmental Stressors published recently by the NATO Advisory Group for Aerospace Research and Development, and which demanded logical reasoning and decision-making functions, memory retrieval functions, and fine manual control. Each task was performed 23 times (6 preflight, 13 inflight, 4 postflight sessions). By means of single subject statistics inflight performance was compared with baseline performance during pre- and postflight sessions. In accordance with the few previous performance studies conducted during space flights, speed and accuracy of short term memory retrieval and logical reasoning functions remained unimpaired during the stay in space. However, clear decrements in tracking performance were found, showing to increased difficulties in fine manual control. These results suggest that performance decrements in space may arise primarily in psychomotor functions due to alterations requiring an effortful accommodation of motor skills which had been acquired under 1-g conditions to the new conditions of microgravity. PMID- 8241724 TI - Illusions of verticality in weightlessness. AB - In weightlessness most subjects feel themselves, and see the visual surroundings, in either an upright or an upside-down orientation although the gravitational force vector is missing. According to a theory of gravity perception, these illusions of positional and visual verticality are assumed to be caused by the force-independent z-axis bias of vestibular and somatic graviceptors. This hypothesis is tested by comparison of measurements of the joint bias in normal gravity with reports of probands in space flight. The expected correlations between the sign of the biases and the occurrence of the respective illusions appear in fact to exist, as well as a negative correlation to incidences of space sickness. If confirmed in a larger sample, the presumed dependency may eventually afford a predictive test of both phenomena. PMID- 8241725 TI - Vestibulo-oculomotor testing during the course of a spaceflight mission. AB - The experimental concept and findings from a recent manned orbital spaceflight are presented. In a single-case, longitudinal study, vestibulo-oculomotor function was examined by caloric testing and active head oscillations. The results from preflight, inflight, and postflight measurements of the human vestibulo-ocular reflex, together with those of ongoing terrestrial studies, should enable separation of the canalicular and otolithic contributions to ocular torsion. This analysis enables an accurate evaluation of the adaptation of the otolithic system to the inflight microgravity and, after landing, to the 1-g force environment. Video-oculography was employed throughout for the comprehensive measurement of eye and head movements. Caloric testing involved air insufflation at 15 degrees C over 90 s, followed by an observation interval of 2 min. During inflight testing this was continued with a 30-s free-floating interval. Active head oscillations were performed at four discrete frequencies (0.12, 0.32, 0.80, 2.0 Hz) and over a frequency sweep between 0.1 and 2.0 Hz. These head oscillations were performed in yaw, pitch, and roll and for three visual conditions (head-fixed target, space-fixed target, no target). The concomitant stimulation of the semicircular canals and otolithic receptors during these oscillations should yield different oculomotor responses under 1-g and 0-g adaptations. Both the short-form caloric test and the active head movement test were performed on 4 of the 5 available mission days. The results of the caloric tests yield a caloric nystagmus intensity (slow-phase velocity) of approximately 60% of that measured before flight and indicate an adaptation in response over the 10-day period after landing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241726 TI - Oculovestibular interactions under microgravity. AB - On a space mission in March 1992 a set of experiments were performed aimed at clarifying the interaction between visual, proprioceptive and vestibular inputs to the equilibrium system. Using the VESTA goggle facility from the European Space Agency we investigated the effect of pure neck receptor stimulation on eye position as measured by the flash afterimage method and on perception of a head fixed luminous line in space. Space vestibular adaptation processes were measured by rotating pattern perception during prescribed head movements. It was found that static ocular counterrotation does not occur under microgravity conditions. This result suggests that the neck receptors apparently do not contribute to a measurable extent. The subjective orientation of a vertical line was perceived correctly inflight. Obviously neck receptors on the perception level can fully substitute for the ineffective equilibrium organs of the inner ear within less than 4 days. The rotating pattern perception during different head motion patterns is not influenced by the absence of a gravity reference. PMID- 8241727 TI - Responses to eccentric rotation in two space-bound subjects. AB - Two subjects were rotated eccentrically in the manner described previously. In contrast to a normal control group, settings of a luminous line to the subjective vertical were almost unrelated to the gravitoinertial vector before, and totally so shortly after, space flight. Only 3 days postflight did a clear relation to the gravitoinertial vector re-establish itself in the one subject who actually flew. The correspondence became normal 5 days after the flight. Since there were no clinical abnormalities evident in the subjects, it is suggested that both subjects suppressed their vestibular information, presumably as an effect of vestibular deconditioning training before the flight. In addition, as a consequence of the flight experience one subject continued to ignore it several days after the flight. PMID- 8241728 TI - Community mental health teams: a suitable case for treatment. PMID- 8241729 TI - Supporting the nurse as well as the patient. PMID- 8241730 TI - Autologous blood for elective oral-maxillofacial surgery. AB - The use of autologous blood in elective surgical procedures is increasing in popularity among surgeons. Initially, it was confined to a small number of specialties, but now it is a technique that nurses are likely to meet during their career. PMID- 8241731 TI - Should practice nurses be involved in diabetic care? AB - Primary care teams who involve practice nurses in the management of patients with diabetes mellitus provide better care. Measures of the structure and process of care are improved when nurses are involved in diabetic care. PMID- 8241732 TI - Humour and the psychotherapeutic process. AB - Communication is the most essential component of psychotherapy. This article aims to define humour, which is one of the most widely used components of communication in day-to-day interaction. PMID- 8241733 TI - Care of a patient undergoing spinal anaesthesia. AB - The preoperative visit by the anaesthetist and/or the preoperative visiting nurse is an important stage in gaining the patient's confidence when explaining a spinal anaesthetic. A brief simple explanation of the procedure is necessary in order to reassure the patient. The patient should be made aware that a general anaesthetic may be required if an adequate block is not achieved. Hypotension is a common side effect following spinal anaesthesia. It is important that intravenous access and fluid therapy are in progress before the spinal anaesthetic is commenced. Psychological and emotional support is essential for patients undergoing spinal anaesthesia, as this can be a frightening experience. A hand to hold is well appreciated. PMID- 8241734 TI - Accreditation and nursing practice. AB - As part of a larger study (Conway, 1993) the literature relating to accreditation of prior learning (APL), accreditation of prior experiential learning (APEL), and the credit accumulation and transfer scheme (CATS) was examined. Significant influencing factors are identified and a general overview of the background to APEL is presented. PMID- 8241735 TI - Nurses are 'demoralized, confounded and confused'. PMID- 8241737 TI - Research and nurse aid. AB - This article asks whether nurses make inadequate first aiders and invites nurses to fill in a questionnaire as part of a research study into nurses' knowledge of first aid and emergency care. The results will be presented in this journal in 1994. PMID- 8241736 TI - Ethics and disability. AB - By using principlism to teach ethics, we are reinforcing a sense of professional superiority among health care professionals. If we are to adequately serve not just the disabled, but all those who come to us for care, we need to rethink and restructure ethics teaching to create a learning environment that promotes and enhances caring. PMID- 8241738 TI - Nursing researchers compromised by contractual obligations. PMID- 8241739 TI - Euthanasia: fact, fear and fantasy. PMID- 8241740 TI - Nursing management of patients receiving a nasogastric feed. AB - When a patient is unable to take a nutritionally adequate diet orally and is malnourished or likely to become malnourished, nasogastric (NG) feeding is normally the first choice of artificial nutritional support. This article describes the initiation of NG feeding and, using a problem-solving approach, explains the nursing care of a patient receiving a NG feed. PMID- 8241741 TI - Preventing childhood accidents: an intervention exercise in Clwyd. AB - Childhood accidents cause much suffering in terms of morbidity and death. There are also enormous financial implications. This article describes a training programme in preventive care that formed the basis for a workshop to increase health care workers' awareness of the problem areas. PMID- 8241742 TI - Health care delivery for people with HIV infection and AIDS. AB - Health care delivery for people with HIV infection and AIDS will need to change in the future to accommodate the expected increasing numbers of people affected. Nurses have an important role in preventing the spread of HIV infection and in caring for this group of people. PMID- 8241743 TI - Ethical dilemmas in caring for a patient refusing amputation. PMID- 8241744 TI - Laura Davies: should children always get the treatment they need? PMID- 8241745 TI - First-aid management of psychological emergencies: 2. AB - Although disasters are not common in our society it is important that all nurses are prepared to help if one occurs in their locality. This article explores what nurse aiders can do in the emergency situation when faced with people suffering psychological reactions to disasters. PMID- 8241746 TI - When and how to seek professional advice from the UKCC. PMID- 8241747 TI - Planning and resource allocation in NHS colleges of health studies: 2. AB - In the second of two articles on the funding of nurse education, the strengths and weaknesses of current funding arrangements for nurse education are explored. The highly regulated health education market is examined and an analysis is offered on the degree to which colleges are able to plan effectively in a competitive and highly unstable market place. PMID- 8241748 TI - Government moves goalposts for community care. PMID- 8241749 TI - Are nurses losing control of care? PMID- 8241750 TI - Health promotion in the elderly. AB - Too often age is equated with pathology. Health promotion strategies need to be modified for the elderly. They must include social and political action as well as life-style changes. PMID- 8241751 TI - Empowerment of diabetics: a challenge for community nursing. AB - Empowerment of diabetic patients can be promoted by the use of blood glucose self monitoring. Community nurses play an important role in teaching effective and accurate techniques. PMID- 8241752 TI - Subjectivity of stress. AB - Stress is a sociopsychophysiological response to environmental stressors. In this article the subjectivity of stressors and the stress response are explored. There is a critical appraisal of physiological and biological theories regarding stress. PMID- 8241753 TI - Management of enuresis in children. AB - A postal survey was conducted to gather information on the enuresis service of Oxfordshire Health Authority Community Unit. This article discusses the size of the problem, treatment and knowledge and expertise. The findings are summarized in the form of a list of recommendations. PMID- 8241754 TI - Taster courses: increasing career choice. AB - Taster courses allow potential students to gain hands on experience in a variety of professional settings. They provide a realistic way of widening entry routes and includes students who are best motivated by vocational courses pathways. PMID- 8241755 TI - Doctors and nurses: dangerous liaisons. PMID- 8241756 TI - Should euthanasia be legalized? AB - Patients are now kept alive far longer than their ancestors could ever have been. In cases of terminal illness where there is an accompanying decline in the patient's quality of life, nurses must be aware of the implications of euthanasia. PMID- 8241757 TI - Nurse-aid management of fractures: 1. AB - Fractures occur in the bones of any age group, even the newborn, and are caused in any number of different ways. This article is the first in a three-part series that will examine the causes and types of fractures, the symptoms and presenting factors, nurse-aid intervention and the healing process. PMID- 8241758 TI - Importance of nursing leadership in implementing change. AB - Change is an aspect of all our lives and nurses are no exception. This article explores the concept of change in nursing and highlights the importance of effective leadership in implementing change. It outlines several change strategies and makes suggestions with regard to choosing an effective approach. PMID- 8241759 TI - Complications in exodontia--accidental dislodgment to adjacent anatomical areas. AB - The authors report 4 cases of accidental dislodgement of teeth to adjacent anatomical areas during extraction. The causes and their prevention are discussed and solutions for the problem are suggested. PMID- 8241760 TI - Three-rooted mandibular molars in patients of Mongolian, Caucasian and Negro origin. AB - A total of 328 periapical x-rays from 105 patients of Mongolian origin, 106 of Negro origin and 117 of Caucasian origin were studied. The Mongolian race showed a greater incidence of three-rooted mandibular molars (15.2% of the Mongolian patients, 7.5% of the Negro patients and 6.8 of the Caucasian patients). There was no statistical difference in relation to sex and the incidence of this extra root. PMID- 8241761 TI - Pre- and post-natal protein deficiency in albino rats. I. Weight progression analysis. AB - Alterations in periodontal tissue were analyzed histologically in rats who had received a protein-deficient diet during intra-uterine life due to inadequate nutrition of the mothers and during the post-natal period due to the continuity of poor nutrition. The animals were weighed at sacrifice: 0, 1, 4, 11, 17, 21, 42, 50 and 105 days. Histological data comparing the undernourished and control groups will be the objective of a future publication. This first part reports the statistical analysis of weight progression of the two groups, concerning not only the mean weight, but also the individual weight of each animal at the various sacrifice periods. Weight progression was gradual and proportional in each group, despite the fact that the control group had accentuated weight gain, deviating, with time, from the weight of the undernourished group of the same age. PMID- 8241762 TI - In vitro study of root canal anatomy of maxillary second premolars. AB - The external anatomy of 435 maxillary second premolars and the internal anatomy of 300 of these premolars were studied in vitro. For the study of the internal anatomy, the teeth were decalcified and cleared. One canal was found in 67.3% of the second premolars, two canals in 32.4% and three canals in 0.3% of these premolars. A total of 90.3% had one root and 9.7% two roots. A predominance of root curvature toward the distal was found in maxillary second premolars with both one or two roots. PMID- 8241763 TI - Photopolymerized resins: surface hardness variation in relation to time of polymerization and setting. AB - The authors studied the superficial hardness of composite resins cured for two different times: 20 and 40 seconds of light exposure. The hardness values were read immediately and 24 hours after polymerization. A significant increase in hardness was observed with 40 s exposure as compared to 20 s. Hardness values were significantly higher at 24 h than immediately after polymerization. PMID- 8241764 TI - The use of Histoacryl in endodontics. AB - The oral environment is one of the most complex systems of the human body. It is populated by a variety of microorganisms, with some of the species still not classified. Tissue adhesives, such as cyanoacrylates, have been used in dentistry and in medicine because of their adhesiveness potential to the human tissue, even in the presence of moisture, their biological compatibility, surface isolation, hemostatic properties and bacteriostatic features. Based on these observations, the authors investigated the use of a tissue adhesive (Histoacryl; Laboratorio Braun, RJ, Brazil) with the purpose of sealing the remaining dentin, especially in endodontically treated molars and premolars. The results strongly suggest that Histoacryl controls micro-leakage of oral fluids at the filling/tooth interface. PMID- 8241765 TI - Guided tissue regeneration versus hemisection in the treatment of furcation lesions. A clinical analysis. PMID- 8241766 TI - Dorsal-ventral polarity in the Drosophila embryo. AB - Embryonic dorsal-ventral polarity in Drosophila is established through a series of successive steps and requires the functions of both maternal and zygotic genes. The graded distribution of the transcription factor dorsal in blastoderm nuclei represents the transition from the maternal to the zygotic program. This results in the activation of specific zygotic genes that act to create the regional pattern along this axis. PMID- 8241767 TI - Formation of germ cells in Drosophila. AB - In Drosophila, germ-cell formation is directed by the posterior pole plasm of the embryo. Genes acting to provide the germ-cell determinant have been identified and characterized, and some are now being manipulated to test their roles. Although recent results largely support the notion of a simple pathway for assembly of pole plasm, complexities are becoming apparent. PMID- 8241768 TI - Transcriptional regulation and spatial patterning in Drosophila. AB - Pattern formation in Drosophila is initiated by a small set of asymmetrically distributed maternal transcription factors that act as graded morphogens along the anterior-posterior and the dorsal-ventral axes of the embryo. Recent progress in the field provides first insight into the molecular mechanisms by which long range positional information in the egg causes a series of localized zygotic transcription factors to position the developmental fate along the blastoderm. PMID- 8241769 TI - Splicing of pre-mRNA: mechanism, regulation and role in development. AB - Over the past year, significant progress has been made in the understanding of how RNA-binding factors may facilitate splice-site selection and spliceosome assembly, and confer fidelity to the pre-mRNA splicing reaction. In addition, a number of studies have revealed a complex network of RNA-RNA interactions in the spliceosome, strengthening the structural and functional parallels between nuclear pre-mRNA splicing and the self-splicing group I and group II introns. These new data further support the idea that pre-mRNA splicing occurs by RNA mediated catalysis and illustrate quite dramatically the dynamic nature of conformational changes in the spliceosome cycle. With respect to tissue-specific pre-mRNA splicing, a number of studies have begun to illuminate mechanisms underlying control of splice-site selection and how so-called 'general' RNA binding proteins, such as heterogeneous nuclear ribonucleoproteins, may be involved in determining different splicing patterns. Finally, an emerging theme involving the role of splicing in development is that differential transcriptional programs can be triggered in different cell types by alternative splicing patterns that generate transcription factor isoforms with different activities or DNA-binding specificities. PMID- 8241770 TI - Segmentation of the Drosophila embryo. AB - Segmentation in Drosophila is a sequential process directed by at least 30 genes that encode various types of proteins, including: many transcription factors; a putative RNA-binding protein; a membrane-associated receptor kinase; several intracellular protein kinases; a number of secreted signaling molecules; and others of unknown function. Although the detailed molecular reactions used to generate the metameric subdivisions of the embryo are not yet understood, a general outline of the processes involved has been described. The manner in which spatial relations in the developing embryo are established can now be described. PMID- 8241771 TI - Evolution and role of Pax genes. AB - Pax genes encode a class of highly conserved transcription factors containing a paired-domain. These factors play important roles in Drosophila and vertebrate development, for example, in segmentation and neurogenesis. Their developmental roles are assessed in terms of their participation in conserved gene networks and mechanisms that establish positional information. PMID- 8241772 TI - Neural fate specification in Drosophila. AB - The specification of cell fates, particularly in the nervous system where cell diversity is highest, is a basic problem in developmental biology. Mutational and molecular analyses in Drosophila are uncovering families of genes, many of them transcription factors, that regulate the progressive acquisition of neural traits. These comprise the initial selection of neural precursors from the ectoderm, the implementation of a basic neural fate common to all precursors and the concomitant endowment of each precursor and its progeny with specific fates. PMID- 8241773 TI - Midline cells and the organization of the vertebrate neuraxis. AB - Vertebrate embryos exhibit a striking midline axis of symmetry that can be recognized in the overall body plan, the framework of skeletal structures and the organization of the nervous system. Cells located at the midline of the embryo during gastrulation have a crucial influence on the establishment of cell identity and pattern within the nervous system. The identification of transcription factors and secreted proteins that are expressed by these midline cell groups has begun to provide a molecular characterization of the organizing centers that establish early neural identity and pattern. PMID- 8241774 TI - Mutational approaches to studying embryonic pattern formation in the zebrafish. AB - The induction and isolation of mutations in genes playing important roles in pattern formation have been extremely valuable in elucidating the mechanisms governing pattern formation in invertebrates. The recent establishment of genetic methods in the zebrafish, Brachydanio rerio, has permitted systematic mutational studies of vertebrate embryonic development. PMID- 8241775 TI - Control of vertebrate gastrulation: inducing signals and responding genes. AB - Recently, genes with similar expression patterns in the early gastrulae of several different vertebrate species have been identified. The remarkable conservation of these expression patterns suggests that fundamental similarities exist within the vertebrates at remarkably early stages. It has yet to be established exactly how these genes are activated in the correct spatial patterns and what their functions might be. PMID- 8241776 TI - The community effect, dorsalization and mesoderm induction. AB - The community effect is an interaction among muscle progenitor cells of amphibian gastrula, and is necessary for the initiation of muscle-specific gene expression. Dorsalization provides a signal that can convert ventral mesoderm cells to a muscle fate. Neither process involves mesoderm-inducing molecules. We suggest that the developmental significance of the community effect is to generate homogeneous but clearly demarcated groups of cells from progenitor cells arranged in a continuous gradient of developmental potential. PMID- 8241777 TI - Pattern formation and developmental mechanisms. PMID- 8241778 TI - Non-analytic problems in detecting arsenic and cadmium in children living near a cadmium refinery in Denver, Colorado. AB - The aim of the present study was to determine urinary arsenic (N = 322) and cadmium (N = 366) levels in children aged six months to six years who live near a working cadmium refinery and to compare their values with those of children from comparison neighborhoods. A questionnaire designed to identify exposure pathways was administered to the parents. There were unexpected problems in the study. Eighty-four percent of the arsenic samples were below the detection limit of 10 micrograms/l and summary statistics could not be calculated. Urinary arsenic and cadmium values could not be standardized for volume and concentration of urine because a large proportion of the samples had very low creatinine values. The original round of cadmium testing was afflicted with contamination problems, possibly due to the mishandling of pediatric urine bags by the parents during the collection procedure. A retest for cadmium levels under clinical conditions showed lower cadmium levels, all but two were below the detection limit. While biological monitoring of exposure to metals can be undertaken indirectly by measuring the concentration of the metals in urine, the analyses in this study were complicated by (1) not using more sensitive analytical tests for arsenic and cadmium determination, (2) not being able to standardize children's urinary values with creatinine, and (3) allowing in-home urine collection. PMID- 8241779 TI - Potential impact of deliberately introduced ozone on indoor air quality. AB - Population subsets whose behavior leads to unusually large exposures to toxicants are of interest to exposure assessors. Despite U.S. Food and Drug Administration (FDA) restriction since 1974, ozone generating devices continue to be marketed in the U.S. for indoor use. Promotional materials cite a host of alleged benefits to indoor environmental quality. Generation rates cited in sales literature are used here to estimate indoor air ozone concentrations that could result from use of such equipment. Predictions exceed relevant air quality standards, in some cases by a substantial margin. Limited available experimental measurements support this finding. PMID- 8241780 TI - The distribution of lifetime cumulative exposures to radon for California residents. AB - The distribution of individual lifetime exposures to radon was estimated using data from studies on radon concentrations, mobility, and time-activity patterns in the state of California. The distributions of radon concentrations in various geographic regions were obtained from the results of year-long radon measurements of 310 residences. The mobility patterns were acquired through a survey of the moving histories of the members of 507 households. The indoor and outdoor time activity data were collected for 1,780 individuals in 1,596 households. Based on these data, a computer simulation technique was used to estimate the distribution of radon exposures with a parametric (lognormal model) and a nonparametric approach (bootstrap method). The estimated average lifetime exposure for radon was 2,448 Bq.m-3.yr for the lognormal model and 2,487 Bq.m-3.yr for the bootstrap method. The standard deviation was 1,130 and 1,145 Bq.m-3.yr respectively. Assuming no move over the lifetime, the estimated average lifetime exposure to radon was 2,052 Bq.m-3.yr for the lognormal model and 2,078 Bq.m-3.yr for the bootstrap method, while the standard deviation increased to 1,378 Bq.m-3.yr for the lognormal model and 1,514 Bq.m-3.yr for the bootstrap method. PMID- 8241781 TI - Chronic disease associated with long-term concentrations of nitrogen dioxide. AB - A prospective epidemiologic cohort study of 6,000 residentially stable and non smoking Seventh-day Adventists (SDA) in California was conducted to evaluate long term cumulative levels of ambient nitrogen dioxide (NO2) in association with several chronic diseases. These diseases included respiratory symptoms, cancer, myocardial infarction (MI), and all natural causes mortality. Cumulative ambient concentrations of NO2 were estimated for each study subject using monthly interpolations from fixed site monitoring stations and applying these estimates to the monthly residence and work place zip code histories of study participants. In addition, a personal NO2 exposure study on a randomly selected sample of 650 people in southern California was conducted to predict total personal NO2 exposure using household and lifestyle characteristics and ambient NO2 concentrations. It was found that good predictability could be obtained (correlation coefficient between predicted and observed values = 0.79) from a model predicting personal NO2. The resulting regression equations from the personal NO2 exposure study were applied to the epidemiologic study cohort to adjust ambient concentrations of NO2. PMID- 8241782 TI - Particle Total Exposure Assessment Methodology (PTEAM) 1990 study: method performance and data quality for personal, indoor, and outdoor monitoring. AB - The Particle Total Exposure Assessment Methodology (PTEAM) study provided the opportunity to test methodologies for measuring personal and microenvironmental PM10 and PM2.5 concentrations in a full-scale probability-based sample of 178 persons and homes in Riverside, California during the fall of 1990. The purpose of the study was to estimate frequency distributions of exposure to PM10, PM2.5, and selected elements in an urban population. Quality control samples and analyses were used to evaluate method performance. These included collocated sample collection, field and lab blank filters, sampler and balance field audits, and intra- and interlaboratory replicate elemental analyses. A portion of the study was also designed to include side-by-side operation of the personal and microenvironmental samplers with reference method (high-volume and dichotomous) samplers to provide an evaluation of method comparability. Over 95% of the approximately 2,900 scheduled samples were collected and analyzed, with very few losses due to equipment failure. The method limit of detection for the personal and microenvironmental monitor PM10 sampling was 8 micrograms/m3. Mean relative standard deviations (RSDs) of 2% to 8% were obtained for collocated personal and microenvironmental samples. Sampler flow rates were within the +/- 10% accuracy criterion during two field audits. Balances operated in a specially designed mobile laboratory were within specified tolerances for precision (+/- 4 micrograms) and accuracy (+/- 50 micrograms). Elemental analysis accuracy was measured with standard reference materials with biases ranging from 2% to 7%. Measurement precision for most elements ranged from 2.5% to 25% mean RSD. Personal and microenvironmental samplers gave median PM10 concentrations that were approximately 9% higher than the dichotomous sampler and 16% higher than the high-volume sampler across 96 monitoring periods at a fixed outdoor location. PMID- 8241783 TI - The history and classification of hydrocephalus. AB - Although great strides have been made toward understanding and managing hydrocephalus, the problem has not been solved completely. This article offers a discussion of hydrocephalus according to historical perspectives, various surgical techniques, including ventriculoatrial and ventriculoperitoneal silicone shunt systems, and classification schemes. PMID- 8241784 TI - The treatment of hydrocephalus. AB - Hydrocephalus is one of the most commonly encountered clinical problems in neurosurgery and accompanies most cases of myelomeningocele. Spinal fluid shunting to the peritoneal cavity is the most common surgery performed for hydrocephalus, and there is a vast array of valve systems and hardware available. The authors present their techniques and refinements for placement of shunt systems and strategies to avoid complications. The medical management of hydrocephalus and shunt infections also is examined. PMID- 8241785 TI - Arrest and compensation of hydrocephalus. AB - We believe that children beyond 5 years of age with stable ventriculomegaly can be monitored if their intellectual performance is within the normal range and stable. The child 3 years old or younger with moderate or greater ventriculomegaly should be shunted. Close follow-up with objective data is essential for all other children with compensated hydrocephalus, as well as for all children with a shunt. PMID- 8241786 TI - Long-term results in the treatment of hydrocephalus. AB - The majority of infants presenting with progressive hydrocephalus are now treated surgically with ventricular shunting. It is difficult to predict the eventual developmental outcome of most treated patients. This article reviews the results of surgical therapy for infantile hydrocephalus with particular attention to neurologic and intellectual outcome. PMID- 8241787 TI - Complications in ventricular cerebrospinal fluid shunting. AB - The development of effective cerebrospinal fluid (CSF) shunts represents a landmark achievement in neurosurgery. This success, however, has been tempered by a high incidence of serious complications that accompany the diversion of CSF. This article examines the various complications of CSF shunting, including proximal, valve, and distal obstruction; infection; and other rare complications, and management of these complications. PMID- 8241788 TI - Ventriculostomy for treatment of hydrocephalus. AB - Ventriculostomy for hydrocephalus can either obviate completely or reduce considerably the complications of cerebrospinal fluid (CSF) shunts. Renewed interest in this procedure has followed the development of flexible and steerable neuroendoscopes, high-quality video cameras, endoscopic surgical instruments, including lasers, and methods of providing computer-guided stereotactic assistance. Ventriculostomy is the preferred treatment for patients with congenital aqueduct stenosis that requires CSF diversion. It is less successful in patients with acquired aqueduct stenosis but is still a treatment option. Interventriculostomy for treatment of intracranial cysts or loculated regions of the ventricles also can obviate or reduce the need for CSF shunts. PMID- 8241789 TI - The neurobiology of normal pressure hydrocephalus. AB - Normal pressure hydrocephalus (NPH) is an intriguing problem in cerebrospinal fluid (CSF) and brain parenchymal physiology. This article presents what is known about its pathophysiology as well as its clinical presentation and treatment. PMID- 8241790 TI - Phase contrast cine magnetic resonance imaging: normal cerebrospinal fluid oscillation and applications to hydrocephalus. AB - Two-dimensional cine phase contrast (2D-cine PC) magnetic resonance (MR) imaging is an imaging technique that sensitizes MR images to velocity changes in a specific direction while cancelling signals from stationary protons and from motion in other directions. This article reviews the technique, conventions, and applications of this imaging technique to hydrocephalus. 2D-Cine PC and additional more rapid MR imaging methods now in development will allow clinicians to measure CSF velocity, pulsation, and bulk flow noninvasively in normal individuals and in patients with a variety of pathologic conditions. It is hoped that these measurements may be correlated with intracranial pressures to develop clinical tests of brain compliance and improved management of patients with hydrocephalus. PMID- 8241791 TI - Diagnosis and treatment of the slit ventricle syndrome. AB - Slit ventricles are a complication of a functioning shunt. This article examines the diagnosis and treatment of slit ventricles and slit ventricle syndrome, the pathophysiology involved, a rationale for treatment, and the direction of therapy in the near future. The authors conclude that determining which type of SVS is present is the first step in treatment and that the best treatment is a strategy aimed at the specific type of SVS that is responsible for the symptoms. PMID- 8241792 TI - Hydrocephalus in the child with dysraphism. AB - Hydrocephalus develops in approximately 85% of patients with myelomeningoceles and appears most frequently to result from obstruction to cerebrospinal fluid (CSF) flow within the posterior fossa subarachnoid space and at the tentorial hiatus, due to posterior fossa crowding from the Chiari malformation. The presenting features of hydrocephalus are legion; although signs and symptoms of intracranial hypertension are most common, many patients exhibit more subtle and confusing signs, such as intellectual deterioration or behavioral changes, or those that mimic brainstem compression from the Chiari malformation or spinal cord dysfunction due to tethering or syringomyelia. Prompt recognition of these signs and symptoms will direct appropriate therapy toward shunt revision and will help the clinician avoid potentially unnecessary and more dangerous procedures. PMID- 8241793 TI - Intraventricular hemorrhage and prematurity. AB - Intraventricular hemorrhage or hemorrhage into the germinal matrix tissues of the developing brain with possible rupture into the ventricular system and parenchyma remains a major problem of preterm neonates and is believed to be attributed to alterations in cerebral blood flow to a damaged germinal matrix capillary bed. Topics discussed in this article include germinal matrix anatomy and physiology, neuropathology, risk of hemorrhage, diagnostic imaging techniques, clinical manifestations, posthemorrhagic hydrocephalus, treatment, and outcomes. PMID- 8241794 TI - Graft-versus-host disease in bone marrow transplantation: experimental, laboratory, and clinical contributions of the last few years. AB - Graft-versus-host disease (GVHD) is a major and often lethal complication of bone marrow transplantation. Research of the past few years has greatly expanded our understanding of the disease and enriched the arsenal of preventive and therapeutic procedures. The present review attempts to give a survey of experimental and clinical GVHD, updating essential knowledge with the latest information until July 1993. The covered topics include the complex immune pathomechanism of acute and chronic GVHD in murine models, the pathogenic role of major, minor, and other antigenic disparities, laboratory markers predicting GVHD, factors influencing appearance and course of the disease, the relationship between GVHD and the graft-versus-leukemia effect, and novel experimental and clinically tested preventive and therapeutic modalities. Finally, the authors set forth their perspective on the most relevant questions in GVHD-related research. PMID- 8241795 TI - Experimental renal transplantation: IgG antibodies which bind to graft tubular epithelial cells cannot be detected in the serum of recipient rats after pre operative transfusion of donor blood. AB - Transfusion of donor blood 7 days prior to renal transplantation between certain rat strain combinations can delay subsequent graft rejection. The mechanism responsible for this increased graft tolerance remains largely unclear. In this paper it is demonstrated that IgG antibodies with affinity for cultured donor renal epithelial cells develop in the plasma of recipient PVG rats within 7 days of transplantation of a DA rat kidney. These IgG antibodies are not specific for kidney cell-restricted antigens as the activity can be completely absorbed using cells which express donor class-I MHC antigens. However, the IgG may contribute to the acute rejection of graft cells by activating antibody-dependent cell mediated cytolytic (ADCC) mechanisms. Transfusion of donor blood 7 days prior to renal transplantation between DA and PVG rats prevented the development of IgG antibodies which bind to resting or cytokine-activated donor renal epithelial cells. Serum taken 7 days after the transplantation of animals which had been previously transfused with donor blood was not able to activate ADCC mechanisms of donor cells. The donor blood transfusion-mediated abrogation of IgG binding to graft cells may delay the onset of graft rejection by preventing antibody dependent graft cell lysis. PMID- 8241796 TI - A decrease in reactive disulfide bonds of serum IgG signals a characteristic change in the IgG subclass patterns of rats bearing experimental tumors. AB - Previous studies have shown that human IgG1 contains a 'reactive' disulfide bridge (SS*), detectable by a 24-hour disulfide exchange reaction, and that the serum level of this IgG subclass is selectively diminished in patients with various malignant diseases. Here we present evidence that in rats IgG2b is the only subclass that carries one SS* per molecule. Furthermore, it is shown that rats inoculated with experimental tumor lines, i.e., the Yoshida hepatoma ascites tumor and the Walker 256 carcinosarcoma growing in ascites or as solid tumor, exhibit significantly decreased SS* per mole IgG which corresponds to a selective diminution of IgG2b. Although at later stages there is a quantitative correlation with the tumor burden, with the Walker tumor this effect becomes significant as early as 24 h after inoculation, i.e., well before exponential tumor growth and an absolute reduction of total IgG. Control animals injected intraperitoneally with either viable spleen cells or irradiated Walker 256 cells did not show comparable alterations in their IgG subclass profile. Thus, the selective defect of IgG2b requires the presence of viable and proliferating tumor cells. Possible mechanism(s) of tumor-associated shifts in IgG subclasses are discussed. PMID- 8241797 TI - IgG subclasses in human hydatid disease: prominence of the IgG4 response. AB - To assess the participation of the four subclasses of IgG in the humoral response to Echinococcus granulosus infection, we determined total and parasite-specific IgG1, IgG2, IgG3 and IgG4 in sera from 46 patients with hydatid disease using an enzyme-linked immunosorbent assay (ELISA). Parasite-specific IgG subclass antibodies were quantitatively measured by means of standard curves obtained by affinity chromatography. Sera from 35 healthy individuals served as controls. The total component of IgG1, IgG2, and IgG3 showed a slight increase in patients with hydatidosis in comparison to normal control subjects with no significant differences. For the IgG4 subclass, however, a marked elevation was found in the patients group (p = 0.001 by analysis of variance). IgG1 and IgG4 subclasses showed a high anti-echinococcus antibody response, whereas there was a low parasite-specific IgG2 and IgG3 response. Indeed IgG-specific antibodies were found to belong mainly to IgG1 (63%) and to IgG4 (30%) and to a lesser extent to IgG2 (5%) and IgG3 (2%). The percentage of the total serum IgG4 antibodies that were specific for hydatid antigen reached a mean level of 18%, significantly higher than that of any of the other three IgG subclasses (p < 0.001 by Student's t test). Thus, the continuous antigenic stimulation of hydatidosis may result in an enhanced IgG4 subclass response. PMID- 8241798 TI - Murine granulocyte-macrophage and mast cell colony formation promoted by nerve growth factor. AB - The nerve growth factor (NGF) is widely distributed in the target tissues of sympathetic neurons. Hemopoietic organs such as the bone marrow (BM) and spleens are known to be innervated by noradrenergic sympathetic neurons. Some of their constitutive cells express NGF receptors (lymphocytes and stroma cells) and its biologic effects have been extensively studied in the immune system and inflammatory responses. However, the effects of NGF on hemopoiesis have been little examined. Recently, we demonstrated that NGF promoted mast cell colony formation from murine BM cells (BMC) or BMC-derived cultured mast cells and induced the phenotypic changes in standard hemopoietic assays. Besides, we demonstrated NGF-enhanced murine neutrophil survival and functional properties. In this study, in order to investigate NGF activities on neutrophil differentiation, we examined granulocyte-macrophage (GM) colony formation from murine BMC or spleen cells (SC) in the methylcellulose culture. We also assessed mast cell colony formation. GM colonies were counted on day 5 and mast cell colonies were counted on day 20 in culture. Although NGF alone (50 ng/ml) neither supported GM nor mast cell colony formation, addition of various doses of NGF to the suboptimal dose of pokeweed mitogen-stimulated SC-conditioned medium (2.5%) or interleukin 3 (50 U/ml), well-known colony-stimulating factors, increased the number of GM and mast cell colonies from both BMC and SC in a dose-dependent manner. These colony formation-enhancing effects of NGF were inhibited by the addition of neutralizing sheep anti-NGF antibodies. The results suggest that NGF may act to develop granulopoiesis including neutrophil and mast cell differentiation in cooperation with hemopoietic factor(s) during inflammatory processes. PMID- 8241799 TI - Eosinophil granulocyte proliferation induced by an intermediate factor generated in the pleural cavity of rats injected with platelet-activating factor-acether. AB - In previous research, we have observed that intrathoracic administration of platelet-activating factor-acether (PAF) promoted a delayed eosinophilia in the pleural cavity of rats that lasted for at least 96 h. We investigated the ability of pleural washings from rats previously injected with PAF (1 micrograms/cavity) to stimulate in vitro murine hematopoietic eosinophil proliferation. We observed that pleural fluid sustained eosinophil proliferation but not differentiation, under conditions in which PAF itself had no effect. The phenomenon lasted for 3 days and was maximal on the 1st day of culture. Treatment with neutralizing antibodies against interleukin (IL)-5, granulocyte-macrophage colony-stimulating factor (GM-CSF) or IL-3, alone or in combination, did not modify the eosinophil proliferation induced by PAF pleural fluid, suggesting that these cytokines may not be involved in the studied phenomenon. We conclude that the rat pleural fluid obtained 6 h after PAF administration induces eosinophil proliferation in vitro by a mechanism probably independent of IL-5, GM-CSF or IL-3. PMID- 8241800 TI - Platelet-activating-factor-induced augmentation of production of eosinophil lineage cells in hematopoietic precursor cells obtained from human umbilical cord blood. AB - Platelet-activating factor (PAF)-induced augmentation of spontaneous differentiation into eosinophil-lineage cells from hematopoietic precursor cells was found among human umbilical cord blood mononuclear cells (CBMCs). This ability of PAF to augment eosinophil production was significantly (p < 0.01) inhibited by the addition of the anti-PAF agent WEB 2086. This enhancing effect of PAF was significantly (p < 0.01) diminished by the presence of antibodies to interleukin (IL)-1 alpha, -beta and IL-3, whereas antibodies to IL-5 or granulocyte-macrophage colony-stimulating factor (GM-CSF) did not diminish it. IL 1 beta, IL-3 or IL-5 gene expression was detected in cellular RNA isolated from both medium- and PAF-stimulated CBMCs using the reverse transcription polymerase chain reaction (RT-PCR) method; however, no expression of GM-CSF was detected. Moreover, 5 times as much IL-3 was detected in the PAF-stimulated CBMC culture supernatant than in that of cells treated with the medium. In the case of IL-1 beta, there was no difference between the 2 cell preparations. On the other hand, no IL-5 or GM-CSF was detected in the supernatant after stimulation with medium or PAF. Depletion of CD2+, CD16+ or CD19+ cells, but not CD14+ cells, from CBMCs resulted in a marked decrease in PAF-induced augmentation of eosinophil production. These results suggest that PAF induces the augmentation of IL-3 production in CBMCs which in turn enhances differentiation into eosinophil lineage cells. PMID- 8241802 TI - Type I allergy to cow milk proteins in adults. A retrospective study of 34 adult milk- and cheese-allergic patients. AB - To determine the clinical and serological characteristics of type I food allergy to cow milk proteins in adults, we investigated all 34 patients (aged from 16 to 58 years) who were diagnosed between 1981 and 1991 to have IgE-mediated reactions to cow milk and cheese. Women represented 91.2% of the study group and 39% of them suffered from the first symptoms during or soon after a pregnancy. So far, 47% of the patients were nonatopic and showed a monovalent sensitization to cow milk proteins. According to RAST results (score > or = 2) the predominant allergen was casein with a sensitization frequency of 71%, whereas sensitization to whey proteins (alpha-lactalbumin and beta-lactoglobulin) in this adult group was rare. In cow milk allergy (CMA) patients, significantly lower levels of milk specific IgG were observed than in normal controls. The main organ manifestations of CMA in adults were the respiratory tract and the skin, with gastrointestinal and cardiovascular symptoms occurring less often than in children. Only 28% of CMA adults could enjoy an unlimited symptom-free intake of milk products after 4 years of disease. Compared with the existing studies on CMA in children, the results suggest that allergies to cow milk proteins in adults are less frequent and tend to persist longer. They are likely to be misdiagnosed for years due to symptoms that initially often refer to only one organ system such as the respiratory tract.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241801 TI - Inhibitory effects of formoterol on platelet-activating factor induced eosinophil chemotaxis and degranulation. AB - A new long-acting beta 2-agonist, formoterol, has been reported to have a greater efficacy and duration of action in asthmatic patients as compared to conventional beta 2-agonists. We recently demonstrated that formoterol inhibited antigen induced late asthmatic response (LAR) and accompanying airway eosinophilia in guinea pigs. In this study, we investigated the direct effect of formoterol in vitro on human eosinophil function, focusing on platelet-activating factor (PAF) induced eosinophil chemotaxis and eosinophil cationic protein (ECP) release. Purified normodense eosinophils were separated by discontinuous gradient from 12 mild asthmatic patients. Formoterol in concentrations of 1-100 microM significantly inhibited PAF-induced eosinophil chemotaxis in a dose-dependent manner with a concentration of drug required to produce 50% inhibition (IC50) of 10.16 microM; % inhibition: 22.9 +/- 13.0% (1 microM), 51.6 +/- 12.7% (10 microM), 75.0 +/- 11.3% (100 microM). When formyl-methionyl-leucyl-phenylamine (FMLP) was used as a chemoattractant, a similar inhibition of eosinophil chemotaxis by formoterol was observed; % inhibition: 13.1 +/- 5.0% (1 microM). 47.7 +/- 7.6% (10 microM), 65.5 +/- 16.5% (100 microM). A conventional beta 2 agonist, salbutamol, at doses to 100 microM did not show any inhibitory effects on PAF-induced eosinophil chemotaxis. Formoterol in concentrations of 1-100 microM also significantly inhibited PAF-induced ECP release from eosinophils; % inhibition: 21.7 +/- 9.0% (1 microM), 39.3 +/- 7.4% (10 microM), 39.6 +/- 8.4% (100 microM). In the presence of phosphodiesterase inhibitors, theophylline or isobutylmethyl xanthine (IBMX), the inhibition by formoterol on PAF-induced ECP release was enhanced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241803 TI - Thrombosis-inducing activity--a factor which appears in plasma of patients with allergic asthma during attack. AB - Thrombosis-inducing activity (TIA) is a factor which was recently identified in plasma from patients with advanced lung cancer and acute respiratory tract infection. In the present study, TIA was measured during periods of attack and nonattack in patients with the allergic and nonallergic type of asthma and pulmonary emphysema with an asthmatic component. In the allergic type of asthma, the attack was accompanied by the appearance of TIA, but when the attack ended it disappeared from plasma. In the nonallergic type of asthma and emphysema with an asthmatic component, TIA remained negative even during the attack. These observations indicate that TIA has some correlations with allergic asthma. PMID- 8241804 TI - Adverse neurologic reactions to the sting of the imported fire ant. AB - While the cutaneous manifestations of the imported fire ant (IFA) sting are well known, neurologic sequelae are much more unusual. We report 2 cases of grand mal seizures associated with the sting of the IFA. The first patient suffered numerous stings without evidence of systemic reactions prior to the onset of the seizure. The second patient had manifestations of anaphylaxis prior to the occurrence of seizure. In both cases, the onset of seizures was delayed. As the geographic range of the IFA increases and more patients are stung, unusual reactions will be seen in greater frequency. PMID- 8241805 TI - Sensitive determination of zearalenone and alpha-zearalenol in barley and Job's tears by liquid chromatography with fluorescence detection. AB - A sensitive and reliable method for liquid chromatographic (LC) determination of zearalenone and alpha-zearalenol in barley and Job's-tears was investigated. The method by which these toxins were determined involves addition of an internal standard (zearalenone 6'-oxime) to barley and Job's-tears samples. Extracts from grain samples were cleaned up by passage through chromatography on piperidinohydroxypropyl Sephadex LH-20 as a lipophilic gel. Individual toxins were resolved by LC on a reversed-phase (ODS) column with fluorescence detection. The detection limit is estimated to be 0.2 ng for zearalenone and alpha zearalenol standards. Known amounts of zearalenone and alpha-zearalenol (25-1250 ng) were added to a barley sample (5 g). Average recoveries for alpha-zearalenol and zearalenone, respectively, ranged from 96 to 102% (mean CV, 3.6%) and from 96 to 103% (mean CV, 3.3%). This method is applicable to determination of alpha zearalenol and zearalenone in barley and Job's tears with satisfactory sensitivity and accuracy. PMID- 8241806 TI - Determination of formaldehyde in fresh and retail milk by liquid column chromatography. AB - A quantitative method is described for the determination of formaldehyde (CH2O) in milk by liquid chromatography. Aldehyde derivatization-extraction was carried out in situ with 2,4-dinitrophenylhydrazine in 2-phase reaction medium. Average recovery of added CH2O (0.1 microgram/mL) was 89.9 +/- 3.9% with an estimated detection limit of 0.009 mg/kg. The technique was used for the determination of CH2O in commercial 2% milk and in fresh milk of cows fed on a typical North American dairy diet. The average concentrations of CH2O in fresh and commercial milk were 0.027 and 0.164 mg/kg, respectively. Several methods for CH2O determination in milk are compared. PMID- 8241807 TI - Simple method for determination of dietary fiber in frozen foods. AB - A simple method was developed for the determination of dietary fiber in multicomponent foods. The method involves dispersing the sample into pH 7.4 phosphate buffer and adding bile and pancreatic enzyme as described. Results were comparable to AOAC methods with correlation coefficients of 86% for multicomponent dinners and 89% for breakfast foods. Coefficients of variation ranged from 7.4 to 20.0% for multicomponent foods and 1.0 to 3.6% for single component foods. In addition, blind duplicate samples had a correlation of 0.99. The described method required less time, labor, and manipulation than AOAC methods. PMID- 8241808 TI - Development of a multispecies identification field test by modified agar-gel immunodiffusion. AB - A multispecies identification field test (MULTI-SIFT) was developed for detection of beef, poultry, pork, sheep, horse, and deer in a wide variety of raw meat products. The test is an adaptation of previously developed field screening immunodiffusion tests for detection of single species. MULTI-SIFT was demonstrated to be specific, relatively sensitive, and accurate in the complete speciation of 100 meat samples. PMID- 8241809 TI - Analysis of milk-based infant formula. Phase IV. Iodide, linoleic acid, and vitamins D and K: U.S. Food and Drug Administration-Infant Formula Council: collaborative study. AB - In 1982, the U.S. Food and Drug Administration, the Infant Formula Council and its member companies, contract laboratories, and other government laboratories began a study of analytical methods for the nutrients listed in the Infant Formula Act of 1980. Phases I, II, III, and V have been completed. The present report provides data on Phase IV, in which 13 laboratories collaboratively studied an ion-selective electrode method for analyzing iodide, a gas chromatographic method for linoleic acid, and 2 liquid chromatographic (LC) methods each for vitamins D and K. Data were insufficient to evaluate one each of the LC methods studied for vitamins K and D. The relative standard deviations (RSD) are sufficient for the nutrient levels found in infant formula. RSDs (%) for repeatability (RSDr) and reproducibility (RSDR), respectively, were as follows: iodide, 4.0-11.4 and 13.5-18.2; linoleic acid, 1.0-1.6 and 3.5-5.1; vitamin K1, 3.2-16.0 and 6.2-19.4; and vitamin D3, 4.2 and 35.0. The recommendation to adopt the method for vitamin D was supported by the results of a ministudy. All laboratories were capable of using these methods with little training. The methods for determination of iodide, linoleic acid, and vitamins D and K in ready-to-feed milk-based infant formula have been adopted first action by AOAC International. PMID- 8241810 TI - A simplified alternative to the AOAC official method for cholesterol in multicomponent foods. AB - An AOAC official method for quantitating cholesterol in multicomponent foods, which was first published in the 13th edition of the Official Methods of Analysis, is rarely used. The method includes so many operations and manipulations--all described in excruciating detail--that most laboratories shun it altogether. Intent on finding an alternative, laboratories have developed their own methods for specific foods. As a result, new methods have proliferated, but still no practical method has been developed for the broader categories of multicomponent foods. The aim of AOAC, which is to promote greater accuracy and uniformity of analytical results primarily through collaborative testing, has not been well served under these circumstances. A different approach guided the work reported in the present paper. This approach was directed toward updating and dramatically simplifying the existing AOAC official method. The method's chloroform-methanol-water mixed-solvent extraction is preserved; however, all the remaining steps have been streamlined, updated, or eliminated by using newer technology. Cholesterol is quantitated with highly specific capillary gas-liquid chromatography using the internal standardization technique. The lipid extract is prepared for the chromatography step by a brief saponification carried out in a culture tube. The resulting method has been validated by using Standard Reference Materials and the standard addition method. Because a simplified method is now available for quantitating cholesterol in the lipid extracts, the expectation is that more attention can be given to the development of improved and efficient extraction methods. This step remains as the central difficulty in any number of methods of analysis for lipid analytes. PMID- 8241811 TI - Enzymatic method for determination of (1-->3)(1-->4)-beta-D-glucans in grains and cereals: collaborative study. AB - The McCleary enzymatic assay for mixed linkage (1-->3)(1-->4)-beta-D-glucans has been modified to apply to oat and barley fractions and ready-to-eat (RTE) cereals. The proposed method involves lower sample concentrations; stirring the samples; a longer, warmer lichenase digestion; and longer beta-glucosidase digestion. These changes result in higher recovery of beta-glucan. This modification expands on the American Association of Cereal Chemists (AACC) Method 32-22 by the addition of a desugaring procedure, which is necessary for RTE cereals. Results from collaborative studies sponsored by AACC and AOAC demonstrate good precision for an enzymatic assay. The average relative standard deviation for reproducibility (RSDR, a measure of interlaboratory variation) for 20 oat, barley, and cereal samples was 9.7%. The average RSD for repeatability (RSDr, intralaboratory variation) was 5.0%. The enzymatic method has been adopted first action by AOAC International for determination of beta-glucans in oat and barley fractions, and ready-to-eat cereals. PMID- 8241812 TI - Methyl bromide determination in selected foods by headspace technique. AB - A headspace method used earlier for determining methyl bromide (MB) in assorted nuts and peanut butters has been successfully applied to other foods that could potentially contain traces of this toxic fumigant. The foods tested include 63 off-the-shelf spices and seasonings, 83 table-ready items (grain-based, dried, or highly seasoned), 30 dried fruits and trail mixes, and 38 oil-based items (oil seeds, cooking oils, or spicy oil-based dressings). Sample headspace gas is produced by blending < or = 50 g sample in 250 +/- 50 mL aqueous solution in a sealed 1000 mL blender cup. After equilibration at 25 degrees C, the headspace is sampled with a gas-tight syringe and injected into a dual column-dual detector gas chromatograph. One determination is made with a 20% OV-101 packed column and a 63Ni electron capture detector (ECD), the other with a GS-Q wide-bore capillary column and a Hall electrolytic conductivity detector (HECD). Of the approximately 200 samples tested, none contained detectable MB residue at a quantitation limit < 100 ng/g sample. All fortified samples yielded MB recovery. Samples were fortified at levels ranging from 78 to 3250 ng MB/g. Recoveries ranged from a mean high of 56% for spices and seasonings to a mean low of 30% for oil-based foods. The overall recovery and CV, including the results from assorted nuts and peanut butters, were 46 and 33%, respectively. PMID- 8241813 TI - Gas chromatographic/electron capture detection method for determination of chlorinated acids in water: collaborative study. AB - A U.S. Environmental Protection Agency (USEPA) interlaboratory method validation study was conducted on USEPA Method 515.1, "Determination of Chlorinated Acids in Water by Gas Chromatography with an Electron Capture Detector." This method is one of the 6 pesticide methods developed for the USEPA National Pesticide Survey (NPS). Method recovery and precision for analyses of sub-ppb to low-ppb concentrations of chlorinated acids were determined in reagent water and finished drinking waters. The analytes evaluated in the study included the 12 pesticides that were quantitatively measured in the National Pesticide Survey (bentazon, 2,4 D, 2,4-DB, 3,5-dichlorobenzoic acid, DCPA-diacid, dicamba, dichlorprop. 5 hydroxydicamba, pentachlorophenol, picloram, 2,4,5-T, and 2,4,5-TP) and 5 pesticides (acifluorfen, chloramben, dalapon, dinoseb, and 4-nitrophenol) that were only qualitatively assessed in the National Pesticide Survey because of recognized method imprecision. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The waters were spiked with 17 chlorinated acids, each at 6 concentration levels, prepared as 3 Youden pairs. Eight laboratories extracted the spiked test waters at pH < 2 with ethyl ether, performed a solvent exchange with methyl tert-butyl ether, prepared methyl esters of the extracted acids using diazomethane, and analyzed an aliquot of each derivatized extract by gas chromatography with electron capture detection. The submitted data were analyzed using a USEPA computer program, which measured recovery and precision for each of the 17 compounds and compared the performance of the method between water types. Method 515.1 was judged acceptable for the 12 NPS analytes recovered quantitatively; mean percent recoveries at 10 15 times the method detection limits ranged from 79 to 105% in reagent water and from 75 to 123% in finished drinking water. In reagent water, overall precision (reproducibility relative standard deviation, RSDR) ranged from 9.6 to 34.2% and in finished drinking water, the RSDR ranged from 11.9 to 37.0%. Single-analyst precision (RSD for repeatability, RSDr) ranged from 5.8 to 17.7% in reagent water and from 4.6 to 27.9% in drinking water. Results for the 5 other NPS analytes were too inaccurate or imprecise and, for these compounds, supported use of the method for qualitative measurements only; the 5 compounds are not included in the adopted method. The method has been adopted first action by AOAC INTERNATIONAL for determination of residues of 12 chlorinated acids in finished drinking water. PMID- 8241814 TI - Gas chromatographic/nitrogen-phosphorus detection method for determination of ethylene thiourea in finished drinking waters: collaborative study. AB - A joint U.S. Environmental Protection Agency (USEPA)-AOAC interlaboratory method validation study was conducted on USEPA National Pesticide Survey (NPS) Method 6, "Determination of Ethylene Thiourea (ETU) in Finished Drinking Water by Gas Chromatography with a Nitrogen-Phosphorus Detector." The purpose of the study was to determine and compare the mean recoveries and precision for determination of ETU in reagent water and finished drinking waters. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The waters were spiked with ETU at 6 concentrations levels, prepared as 3 Youden pairs. In the method, the test water is extracted by passing the sample through an absorbent matrix type tube. ETU is recovered from the tube with methylene chloride, the extract is solvent-exchanged to ethyl acetate, and an aliquot of each extract is analyzed by gas chromatography using a nitrogen-phosphorus detector. Twelve laboratories participated in the study. Data were analyzed using a USEPA computer program, which measured recovery and precision for ETU and compared the performance of the method between the 2 water types. Over the concentration range tested, the mean percent recoveries of ETU were 82-92% in reagent water and 85-98% in finished drinking water. The range of the between laboratory relative standard deviations (RSDR) for the 6 concentrations was 5-24% in reagent water, but was only 4-9% in finished drinking water. The range of the within-laboratory relative standard deviations (RSDr) was 6-14% for reagent water and 6-10% for finished drinking water. Results for the 2 water matrixes showed no statistically significant differences.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241815 TI - Determination of pesticides in indoor air and dust. AB - Improved analytical and sampling methods were developed for the multiresidue determination of pesticides in indoor air. Air analysis consists of adsorption of the pesticides in 1 m(3) of air onto Tenax TA via an air sampling pump, desorption with acetone and determination and quantitation by gas chromatography/mass spectrometry (GC/MS) with chemical ionization on an ion trap mass spectrometer. Limits of detection for the 23 pesticides studied ranged from 0.5 ng/m(3) for chlorpyrifos and diazinon to 30 ng/m(3) for o-phenylphenol (approximately 0.5-30 parts per trillion on a w/w basis). A simple method for the detection of pesticides in dust was also developed. This method involves emptying the contents of a vacuum cleaner bag into a standard household food processor and extracting 1 g homogenized dust with acetone before GC/MS. Limits of detection were 25-100 ppb because of interferences by common household chemicals. However, pesticide concentrations were higher in dust than in air, therefore, analysis of dust is a better indicator of indoor pesticide exposure. PMID- 8241816 TI - Gas chromatographic determination of cyclohexanone leached from hemodialysis tubing. AB - A capillary gas chromatographic procedure is described for the determination of cyclohexanone leached from hemodialysis tubing by water. Recoveries were 100% at 2.0 mg/100 mL (20 ppm), 99.0% at 500 microg/100 mL (5 ppm), and 106% at 1.0 microg/100 mL (10 ppb). Reproducibility of the system was 0.152% for 3.0 microL injections of a solution containing cyclohexanone at 5.58 microg/mL. Correlation coefficients were 0.9983 for 0.3672-3.672 ng and 1.0000 for 3.672-367.2 ng. Twenty hemodialysis tubing sets from 4 manufacturers were examined. The leachable cyclohexanone ranged from 1.02 to 43.7 ppm per set. Rinsing the tubing with 1 L 0.9% sodium chloride solution did not remove significant amounts (P = 0.05) of leachable cyclohexanone. PMID- 8241817 TI - Determination of 2-ethyl-1-hexanol as contaminant in drinking water. AB - An analytical procedure for the determination of 2-ethyl-1-hexanol (2-EH) in drinking water is presented. The method is based on volatile-compound stripping, adsorption on activated-charcoal-filled tubes, solvent elution, identification by gas chromatography/mass spectrometry, and determination by gas chromatography with flame ionization detection. Bottled samples with undesirable organoleptic characteristics were analyzed to determine a possible correlation with the presence of 2-EH. The presence of 2-EH at 2-10 micrograms/L was confirmed in several samples. The presence of di-2-ethylhexyl phthalate (D2EHP) was also checked in all samples. This compound was always found at 2-30 micrograms/L. Hydrolysis of D2EHP was carried out for 2 weeks to evaluate its possible contribution to water contamination by 2-EH. Tests did not show measurable amounts of the alcohol. Nonetheless, the hydrolysis of phthalates in the weakly acidic conditions of the examined waters would not justify the presence of 2-EH at the observed levels, and so it is reasonable to hypothesize a direct contamination from packaging materials containing 2-EH as residue from D2EHP synthesis. PMID- 8241818 TI - Determination of methylene chloride acceptability and its "purification" for ethylenethiourea methodology. AB - A liquid chromatographic-electrochemical method for the determination of ethylenethiourea (ETU) residues uses methylene chloride in the cleanup. Distilled in-glass grade methylene chloride produced erratic ETU recoveries ranging from 0 to 106% for vacuum rotary evaporation of the solvent. ETU recoveries were found to be dependent on the bottle of methylene chloride used, not on supplier or lot. When GC2 grade methylene chloride from Burdick & Jackson Laboratories was used, ETU recoveries ranged from 92 to 110%. "Acceptable" ETU recoveries were defined as those values between 90 and 110%. Passing "unacceptable" methylene chloride through a column containing anhydrous sodium sulfate, sodium carbonate, and alumina was found to adequately purify methylene chloride. Treated methylene chloride provided acceptable ETU recoveries for up to 1 month after "purification." PMID- 8241819 TI - Modified method for the determination of ivermectin residues in animal tissues. AB - A simple, reliable method for determining residues of ivermectin in animal tissues is described combining existing sample cleanup procedures for muscle and for feed. Sample extracts are purified on alumina, C18, and silica solid-phase extraction cartridges, and ivermectin is determined by liquid chromatography with fluorescence detection. The average recovery of ivermectin from cattle, swine, and sheep muscle and liver was 87%, in the range of 5-25 ppb. Recovery from fish muscle was 77%, in the range of 5-30 ppb. The detection limit in tissue is 2 ppb. PMID- 8241820 TI - Simultaneous liquid chromatographic determination of thiamine and riboflavin in selected foods. AB - A reliable, improved liquid chromatographic (LC) method has been developed for the measurement of thiamine and riboflavin in foods. The major improvement in the method is the chromatographic separation achieved. The method is also very reproducible and extremely sensitive. After autoclave extraction, samples are derivatized to form thiochrome (a highly fluorescent oxidation product of thiamine). Riboflavin is naturally fluorescent. Interferences are removed on a C18 cartridge and chromatographed by using a reversed-phase separation. The mobile phase used is 72% 0.005M NH4OAc (pH 5.0)-28% MeOH. Fluorescence detection using wavelength switching, 370-435 for thiamine and 370-520 for riboflavin, allows determination of each vitamin at its optimum wavelength for maximum sensitivity. Detection limits were 0.05 ng for both thiamine and riboflavin. The method can also be performed by using a fluorescence detector at a single wavelength, but with a sacrifice of sensitivity. Data comparisons between AOAC fluorometric and LC results were excellent for routine samples, as well as for American Association of Cereal Chemists (AACC) check samples. All LC results from AACC samples were within 2 standard deviations of the mean. Reproducibility was 1.9% for thiamine and 1.6% for riboflavin. PMID- 8241821 TI - FDA Monitoring Program. AB - Under regulatory monitoring, 16,428 samples of domestically produced and imported food from 94 countries were analyzed for pesticide residues in 1992. Of these, 15,370 were surveillance samples, which are collected when there is no evidence of a pesticide problem. No residues were found in 65% of the domestic surveillance samples and 66% of the import surveillance samples. Findings in the 1058 compliance samples reflect their collection and analysis when a pesticide problem is suspected. Under incidence/level monitoring, 206 samples of aquaculture seafood/shellfish and 558 milk samples were analyzed for pesticide residues. The findings were similar to those from FDA's regulatory monitoring. The 1986-1991 Total Diet Study average results (dietary intake of pesticides) over that period are generally well below the standards set by the JMPR and by EPA. These 5-year averages may differ from the previously reported 1-year intakes (6-10) because they cover a period during which intakes of some chemicals, such as persistent chlorinated pesticides, have been generally declining, and also because they represent a 5-year rather than a 1-year average. PMID- 8241822 TI - The need for larger analytical samples with granulated feed additives. AB - Sampling an animal feed containing a granulated analyte is extremely difficult. A granulated feed additive, Tylan (Elanco, tylosin), was used to demonstrate how a granulated product can increase assay variability if steps are not taken to reduce and control this variability. The number of tylosin granules was determined for Granulated Tylosin Concentrate, the active ingredient of Tylan Premix. Multiple weighings of 3 lots of the concentrate were prepared, and the average number of granules per gram of concentrate was determined with the aid of image analyses. The total weight of Tylan Premix in varying analytical samples was calculated. Finally, the average number of tylosin granules per analytical sample was calculated based on the average number of granules per gram. A 10 g analytical sample obtained from an animal ration containing 8 ppm tylosin activity would contain an average of 1.16 granules of tylosin, and a 100 g analytical sample would contain 11.6 granules. These calculations demonstrate the need to increase the analytical sample size in analyzing an animal feed containing a granulated feed additive. PMID- 8241823 TI - Solubility of antibiotics used in animal feeds in selected solvents. AB - The solubility of antibiotics used in animal feeds in organic solvents was determined. The solubility data could be used in identification of classes and in some cases individual members of the same class of antibiotics, and in differential step(s) in the analysis of these antibiotics. A universal solvent was developed to extract all antibiotics from animal feeds. PMID- 8241824 TI - Determination of Roxarsone in feeds using solid phase extraction and liquid chromatography with ultraviolet detection. AB - A method is presented for detection and quantitation of Roxarsone in poultry feed by liquid chromatography. The drug is extracted by phosphate buffer and determined by solid phase extraction and reversed-phase liquid chromatography. Recoveries of the sample spikes and fortified field samples agree closely with those obtained by the standard spectrophotometric method. PMID- 8241825 TI - Liquid chromatographic determination of six sulfonamide residues in animal tissues using postcolumn derivatization. AB - A sensitive and reproducible liquid chromatographic method is described for the simultaneous determination of sulfonamide residues in animal livers and kidneys. The selectivity of the method is enhanced significantly through the use of postcolumn reaction with p-dimethylaminobenzaldehyde, followed by detection at 450 nm. Consequently, the cleanup is simplified; it consists of removal of fatty material by partitioning the sample extract into an acetonitrile-hexane system. The sulfonamides used for this study were sulfadiazine, sulfapyridine, sulfamerazine, sulfadimidine, sulfamethoxypyridazine, and sulfaquinoxaline. At the level of 100 micrograms/kg, the recoveries ranged from 70 to 104% and were dependent on the nature of the matrix and the particular sulfonamide. The coefficients of variation are 2-10% at this level. The detection limit was 20 micrograms/dg. PMID- 8241826 TI - Confirmatory method for sulfonamide residues in animal tissues by gas chromatography and pulsed positive ion-negative ion-chemical ionization mass spectrometry. AB - A confirmatory method has been developed for determination of 13 sulfonamides in edible tissues. The assay involves extraction from a solution resulting from a screening procedure by liquid chromatography and subsequent derivatization. Sulfachloropyridazine (SCP), sulfadiazine (SDA), sulfadimethoxine (SDM), sulfamethazine (SMZ), sulfamerazine (SME), sulfamethoxazole (SMX), sulfamethoxypyridazine (SMP), sulfapyridine (SPR), sulfaquinoxaline (SQX), and sulfathiazole (STA) were detected as the N1-methyl-N4-trifluoroacetyl derivatives, sulfaguanidine (SGU) as the same derivative after cyclization by hexafluoroacetylacetone, and sulfacetamide (SAC) as the methyl derivative. These sulfonamides were detected by gas chromatography and pulsed positive ion-negative ion-chemical ionization mass spectrometry with methane as the reactant gas, whereas sulfanilamide (SAA) was determined as the methyl derivative by electron impact ionization. PMID- 8241827 TI - Variability among peanut subsamples prepared for aflatoxin analysis with four mills. AB - The variability in aflatoxin concentration among peanut subsamples ground with 4 different mills was evaluated. Twenty 2 kg samples of naturally contaminated peanuts were ground in a Dickens subsampling mill (DM), a Stephan model UM-12 vertical cutter mixer (SM), and a Robot Coupe model RS16Y-1 vertical cutter mixer (RC1). Twenty 4 kg samples were ground in the DM, SM, and a Robot Coupe model R10P vertical cutter mixer (RC2). From each 2 kg sample, ten 100 g subsamples were withdrawn, and from each 4 kg samples, ten 200 g subsamples were withdrawn. Subsamples were analyzed for aflatoxin by liquid chromatography. The coefficient of variation (CV) among each set of 10 subsamples was determined for each sample, and the CVs for each sample size were ranked and analyzed by the Kruskal-Wallis test of ranks. For 2 kg samples, the CVs for the samples ground in RC1 ranked significantly lower than those for samples ground in DM and SM. For 4 kg samples, the CVs for samples ground in RC2 and SM were significantly lower than that for samples ground in DM. The averages of the CVs for 2 kg samples were 17.2% (RC1), 32.8% (SM), and 40.6% (DM). The averages of the CVs for 4 kg samples were 21.2% (RC2), and 26.0% (SM), and 47.0% (DM). PMID- 8241828 TI - Substrate supporting disc method for confirmed detection of total coliforms and E. coli in all foods: collaborative study. AB - The ColiComplete substrate supporting disc (SSD) method for simultaneous confirmed total coliform count and Escherichia coli determination in all foods was compared with AOAC most probable number (MPN) methods, 966.23 and 966.24. Twenty-nine laboratories participated in this collaborative study in which 6 food types were analyzed. Four food types, raw ground beef, pork sausage, raw liquid milk, and nut meats, were naturally contaminated with coliform bacteria. Two foods, dry egg and fresh frozen vegetables, were seeded with coliforms. Three food types, ground beef, raw liquid milk, and pork sausage, were naturally contaminated with E. coli. Although pork sausage was naturally contaminated, the level was very low (<10/50 g); therefore, additional E. coli were inoculated into 1 lot of this food type. Three food types, nut meats, dry egg, and fresh frozen vegetables, were inoculated with E. coli. For naturally contaminated samples, duplicate determinations were made on 3 separate lots for each food type. For inoculated samples, low, medium, and high contamination levels plus uninoculated control samples were examined in duplicate. Data were analyzed separately for total coliform bacteria and for E. coli. Mean log MPN counts were determined by the SSD method and the appropriate AOAC MPN method. Results were then analyzed for repeatability, reproducibility, and mean log MPN statistical equivalence. Results were statistically equivalent for all total coliform levels in all food types except frozen vegetable and raw nut meat uninoculated control samples and 1 lot of pork sausage where the SSD method produced statistically significant greater numbers. For the E. coli determinations, results were statistically equivalent across all samples and all levels for each food type. The SSD method has been adopted first action by AOAC International for confirmed detection of total coliforms and E. coli in all foods. PMID- 8241829 TI - Management of sickle cell disease; lessons from the Jamaican Cohort Study. AB - Sickle cell disease is enormously variable in its expression and outcome. In addition to this intrinsic variability are the problems of symptomatic selection biasing observations towards the severe end of a wide clinical spectrum and a truly changing natural history as a result of better management. Against this background, there was a need for a description of the disease in a truly representative sample of patients and this objective has been approached in the Jamaican Cohort Study of Sickle Cell Disease. Initiated in 1973, this study is based on all cases of sickle cell disease detected among 100,000 consecutive normal deliveries in Kingston, Jamaica. All affected children as well as age matched normal controls have been followed prospectively and are currently aged 11 to 19 years. The following review is based on lessons learnt from this cohort study. It is not intended to be a comprehensive survey of knowledge of sickle cell disease and does not address major contributions from studies elsewhere. In some ways, therefore, the review may appear unbalanced because of this specific objective. However, a great deal has been learnt about the evolution of the abnormal haematology of sickle cell disease and its relationship to clinical features. The causes of early mortality in sickle cell disease in Jamaica are described and the major complications such as acute splenic sequestration, pneumococcal septicaemia, aplastic crisis, hypersplenism, and acute chest syndrome have been addressed with varying success. Overall survival to the age of 19 years has been 75% and it is planned that the study should continue to define the problems of late adolescence and early adult life. PMID- 8241830 TI - The unstable haemoglobins. AB - The unstable haemoglobin haemolytic anaemias result from the presence in the red cell of a structurally abnormal haemoglobin variant. There are many mutations producing unstable haemoglobins; most are single amino acid replacements that affect a few key areas of the haemoglobin structure. A wide range of haemoglobin instability is evident from in vitro studies, extending from mutants with a subclinical degree of instability to those associated with severe haemolytic disease. The characteristic feature of variants associated with haemolysis is a markedly decreased stability which is readily detectable by simple screening tests. The in vivo consequence is the precipitation of the unstable haemoglobin to give Heinz bodies which are associated with the red cell membrane and lead to premature cell destruction. The unstable haemoglobins have a greater tendency to spontaneously oxidise to methaemoglobin with subsequent formation of haemichromes and precipitation. This process is significantly accelerated by external factors such as exposure to oxidative substances and increased temperature; thus haemolytic crises are frequently associated with infections in otherwise asymptomatic carriers of unstable haemoglobins. The clinical expression of the unstable haemoglobin mutation may also be modified by proteolysis of the unstable globin chain in the bone marrow. This proteolytic mechanism can predominate in the case of extremely unstable globin chains to produce primarily a thalassaemic phenotype with little if any circulating unstable haemoglobin or evidence of haemolysis. PMID- 8241831 TI - Chronic lymphocytic leukaemia treatment. AB - Advances in the understanding of the natural history and prognosis of chronic lymphocytic leukaemia (CLL) and the introduction of new drugs and strategies have renewed interest in the treatment of this disease. Clinical trials in patients stratified according to risk factors are useful to determine the efficacy of different treatments. These advances currently allow treatment of CLL patients to be decided on a more rational basis. Patients in early clinical stage (Binet A, Rai 0) with non-diffuse bone marrow histopathology, and low, stable blood lymphocyte levels have a long survival and should not be treated unless the disease progresses. On the contrary, patients with poor prognostic features, such as advanced clinical stage (Binet B, C; Rai III, IV), diffuse bone marrow infiltration or high and rapidly increasing blood lymphocyte levels have a median survival inferior to 5 years and should be treated. Chlorambucil is still the mainstay for treatment of CLL but new agents such as fludarabine and 2 chlorodeoxyadenosine offer great promise. These agents are already the treatment of choice for patients failing standard therapies and its role as front-line therapy is being investigated in randomized trials. Whatever the treatment used, however, cure is rarely achieved. A number of situations (e.g. autoimmune cytopenias, hypersplenism) merit special treatment approaches (e.g. corticosteroids, splenectomy). Bone marrow transplants, albeit experimental in CLL, warrant investigation in younger patients with poor prognosis. PMID- 8241832 TI - Action of snake venom components on the haemostatic system. AB - Among the components in snake venom are a number which have profound effects (either stimulatory or inhibitory) on haemostatic mechanisms, including coagulation, fibrinolysis, platelet function and vascular integrity. As a consequence, human victims of snakebite may suffer severe and sometimes fatal haemorrhagic and/or thrombotic sequelae. Many of these venom components have been isolated and their precise mechanisms of action established. Apart from direct fibrinolysins, procoagulants predominate, most of these exerting their effect late in the clotting cascade, activating factor X or prothrombin or directly converting fibrinogen to fibrin. Some of the procoagulants are, or have the potential to be, used as therapeutic agents. Some venom components have been put to use as laboratory reagents for diagnostic purposes or for characterising molecular defects of haemostasis, although because they often have unphysiological actions, results must be interpreted with caution. These and other useful constituents e.g. protein C activator and platelet aggregating agents are discussed. PMID- 8241833 TI - Neisseria meningitidis: serotyping and subtyping by whole cell ELISA. AB - Twenty strains of Neisseria meningitidis isolated from clinically diagnosed cases of meningococcal disease were subjected to serogrouping, employing slide agglutination followed by serotyping and serosubtyping by whole cell ELISA using monoclonal typing antisera. All isolates were from sporadic cases of meningitis during a period of two years from various hospitals in Delhi. All 20 isolates were grouped as serogroup A and typed as serotype 4, except one strain which was untypable. On serosubtyping the isolates were found to belong to P1.9 (7 strains) followed by P1.1 (5), P1.9 (2), P1.16,1 (2), P1.6,10 (2), P1.10,7,1 (1) and non subtypable (1). PMID- 8241834 TI - Dietary intake of Al, Ca, Cu, Fe, Pb, and Zn in infants. AB - The dietary intake of six elements--Al, Ca, Cu, Fe, Pb, and Zn--was measured in 39 normal healthy children aged 17-61 (mean 35.5) weeks. There was a downward trend with age in daily intake of fluid, Pb and Fe, in contrast to an increase in solid intake, Ca and Zn throughout the study. The geometric mean total daily element intake (mg) was: Al 1.12; Ca 446; Cu 0.6; Fe 4.5; Pb 0.020; Zn 2.8. The geometric mean daily element intake (mg) from milks only was as follows: Formula milk (n = 14): Al 0.26; Ca 184; Cu 0.24; Fe 1.85; Pb 0.005; Zn 1.6. Cows' milk (n = 6): Al 0.27; Ca 384; Cu 0.04; Fe 0.21; Pb 0.004; Zn 1.3. Breast milk (n = 12): Al 0.77; Ca 116; Cu 0.15; Fe 0.38; Pb 0.012; Zn 0.4. Additionally, levels of aluminium were investigated in some packaged infant foodstuffs, including four soya milk formulae. The soya-based formulae had a mean concentration of 1.09 +/- 0.37 micrograms/g, which was not significantly different from the mean of 0.83 +/ 0.28 microgram/g for six cows' milk formulae. PMID- 8241835 TI - Measurement of glutathione S-transferase pi by radioimmunoassay: elevated plasma levels in lung cancer patients. AB - Human glutathione S-transferase pi was purified from placenta, and when compared to previous procedures a high yield of pure protein was obtained. A specific and sensitive radioimmunoassay suitable for the measurement of human glutathione S transferase pi in human plasma is described. The sample collection method for glutathione S-transferase pi was found to be a major obstacle in utilising the measurements for diagnostic use, unless a stringent collection protocol was adhered to. A good discrimination between cancer and non-cancer patients was observed, with 66% of lung cancer patients having elevated glutathione S transferase pi levels with adenocarcinomas showing the most significant rises. PMID- 8241836 TI - Cardiac transplantation: a review. AB - Cardiac transplantation is no longer an experimental procedure, but rather a definitive therapeutic option for patients with end-stage cardiac failure. The current 5-year and 10-year survival rates following cardiac transplantation are 67% and 52%, respectively. Rejection and infection still result in significant morbidity and mortality, and the development of graft coronary occlusive disease limits even longer-term survival. This review summarises donor and recipient selection criteria, the operative procedure, the current immunosuppressive protocols, as well as newer immunosuppressive agents and the complications of cardiac transplantation, namely infection, rejection, nephrotoxicity and graft coronary occlusive disease. PMID- 8241837 TI - Ischaemic heart disease, cardiac surgery and heart/heart-lung transplantation reviewed from a haematological perspective. AB - Ischaemic heart disease, one of the major causes of death in the western world, seems to involve a complex interaction of haematological, biochemical, immunological, and physiological processes. Haematologically, the process involves alterations in the haemostatic mechanism as a result of elevations of certain clotting factors, particularly factor VII and fibrinogen, possibly as a result of increased fat consumption and smoking. Platelet hypersensitivity may also be related to immunological damage of the coronary endothelium and elevated fibrinogen levels. Cardiac surgery and heart transplantation remain the most effective methods for treatment of end-stage ischaemic heart disease. Surgical techniques are complex and necessitate the establishment of an extracorporeal blood circulation to bypass the functions of the heart and lungs, enabling the heart to be temporarily paralysed, and allowing surgery to be performed. Extracorporeal oxygenation and circulation of blood, and implantation of ventricular assist devices and artificial hearts, result in exposure of blood to foreign surfaces, leading to activation of the haemostatic mechanism and concomitant haemorrhagic/thrombotic complications. Heart and heart/lung transplantation require long-term regular monitoring of infection and rejection episodes in addition to control of bleeding complications sometimes experienced in the immediate post-operative period. Early identification of lymphoproliferative disorders which may arise as a result of excessive immunosuppression is also important. Implantation of artificial hearts and ventricular assist devices requires much more sophisticated haemostatic monitoring than routine cardiac surgery, in order to control the haemorrhagic/thrombotic complications so often associated with these procedures. PMID- 8241838 TI - Platelet activation in coronary artery disease. AB - The role of platelets in coronary artery disease has become increasingly apparent in the last decade. Platelets are the main components of arterial thrombi, and they also contribute to the development of atheroma and smooth muscle hypertrophy in arterial walls. Platelets are known to circulate as round, inactive spheres which undergo a succession of changes before they become functional. These changes result in activated platelets. Platelet activation has been demonstrated during acute coronary events. There is increasing evidence that an increased proportion of platelets circulate in an activated form in patients with coronary artery disease and also in other disease states in which thrombosis is a feature. In this review we examine some of the evidence for these statements. PMID- 8241839 TI - Atherosclerosis and coronary heart disease. AB - Coronary heart disease is the largest cause of morbidity and mortality in the UK. The disease develops by the interaction of a variety of environmental agents in people who may be genetically susceptible. Some of the environmental agents can be favourably altered by adjustments to lifestyle, particularly by stopping smoking, increasing exercise levels and controlling the diet. Coronary heart disease is manifested by a number of symptoms due to the partial or complete occlusion of the coronary arteries by a fibrous plaque. The processes of plaque formation can begin very early on in life and progress through various stages. The process may be initiated by injury to the endothelial lining but can also occur in areas without obvious injury. Diagnosis of coronary heart disease is largely by electrocardiographic measurements and a variety of imaging modalities and laboratory investigations. Treatment centres upon the medical control of the symptoms, or surgical intervention in the form of a coronary bypass operation or by coronary angioplasty. PMID- 8241840 TI - Laboratory aspects of infective endocarditis. AB - The clinical laboratory has a major role in both the diagnosis and treatment of cases of infective endocarditis. Patients often present with vague non-specific symptoms, which complicates the initial diagnosis. Blood cultures are by far the most important specimens. Laboratories should be aware of isolation problems associated with certain organisms. If initial blood cultures are negative then serological tests for non-bacterial causes should be considered. Staphylococci and streptococci account for > 90% of cases, and appropriate antimicrobial chemotherapy for the majority of strains is well established. The laboratory role is to identify the infecting organism, to establish the in-vitro activity of the proposed antibiotic therapy by determining minimum inhibitory concentrations and minimum bactericidal concentrations of antibiotics, singly and in combination against the patient's own organism, and to measure the bactericidal activity of antibiotics in the patient's serum to assess potential clinical efficacy. Laboratory aspects of diagnosis, treatment and therapeutic monitoring are discussed. PMID- 8241841 TI - Enteroviruses and heart disease. AB - Enteroviruses are ubiquitous, and most individuals encounter numerous serotypes during their lifetime. Infection of the heart is an unusual occurrence but enteroviruses are nevertheless the commonest viral agents of acute myocarditis. The introduction of new serological and molecular virological detection methods has improved the diagnosis of acute enteroviral heart disease and has also provided information on its epidemiology and pathogenesis. There remains, however, much debate on whether cardiac damage results directly from viral infection of myocytes and the ensuing immune response, or indirectly from autoimmune mechanisms. Enteroviruses are also associated with chronic cardiac disease, most notably chronic dilated cardiomyopathy. Recently the strength of this associated has been somewhat weakened by conflicting experimental evidence, but this is still an open question. Further studies are required to delineate the true role of enteroviruses in acute and chronic myocardial disease, to understand its pathogenesis more completely, and to apply this knowledge to the development of preventive or therapeutic treatment regimens. PMID- 8241842 TI - Tissue typing, heart and heart-lung transplantation. AB - The effects of HLA matching and the presence of pre-existing anti-HLA antibodies, together with cross-match results, on heart and heart-lung transplants are discussed. Prospective HLA matching of donor and recipient is not usually performed for heart and heart-lung transplants. In a multi-centre study statistically significant improved survival with the better-matched heart grafts was found. In a small series of heart grafts it was also found that the improvement with HLA matching was confined to male recipients who were not blood group O. Heart graft recipients with wide panel reactive antibody had worse graft survival one year after transplant than the unsensitized. In several studies patients with a positive cross-match had a significantly lower survival rate than those with a negative cross-match. It is suggested that, minimally, pre transplant screening and prospective cross-matching of highly sensitised patients is necessary. PMID- 8241843 TI - Lipoprotein oxidation: the 'fruit and vegetable gradient' and heart disease. AB - Coronary heart disease is a major cause of premature death in Western societies. Oxidation of low density lipoproteins by oxygen free radicals provides a molecular link to the development of atherosclerosis. Free radical oxidations can usually be protected against by appropriate antioxidants. Recent studies suggest epidemiological correlations exist between the levels of plasma antioxidant vitamins and mortality from heart disease. PMID- 8241844 TI - Diabetic control and complications. PMID- 8241845 TI - Routine measurement of fibrinogen concentration. PMID- 8241846 TI - Antiphospholipid syndrome. PMID- 8241847 TI - Electromagnetic fields and childhood cancer. PMID- 8241848 TI - Free radicals and vascular disease: how much do we know? PMID- 8241849 TI - Alternative career paths for doctors. PMID- 8241850 TI - Residence near high voltage facilities and risk of cancer in children. AB - OBJECTIVE: To investigate whether residence before and after birth near 50 Hz high voltage installations increases a child's risk of cancer and whether risk correlates with the strength of the magnetic field. DESIGN: A population based case-control study. SETTING: Denmark. SUBJECTS: 1707 children under the age of 15 with leukaemia, tumour of the central nervous system, or malignant lymphoma diagnosed in 1968-86 and 4788 children taken from the central population register. MAIN OUTCOME MEASURES: Proximity before and after birth to existing or former 50-400 kV electrical transmission connections and substations and associated historical electromagnetic fields calculated on the basis of current load on line, phase ordering of line, and distance from the dwelling. RESULTS: A significant association was seen between all major types of childhood cancer combined and exposure to magnetic fields from high voltage installations of > or = 0.4 microT (odds ratio 5.6). At > or = 0.25 microT no significant association was seen (odds ratio 1.5). A possible association was also seen with cases of Hodgkin's disease separately at > or = 0.1 microT. CONCLUSIONS: On the basis these results and additional descriptive data on electricity consumption and incidence of childhood cancer in Denmark since the 1940s it was concluded that the proportion of childhood cancer possibly caused by 50 Hz electromagnetic fields must be small. PMID- 8241851 TI - Risk of cancer in Finnish children living close to power lines. AB - OBJECTIVE: To investigate the risk of cancer in children living close to overhead power lines with magnetic fields of > or = 0.01 microteslas (microT). DESIGN: Cohort study. SETTING: The whole of Finland. SUBJECTS: 68,300 boys and 66,500 girls aged 0-19 years living during 1970-89 within 500 m of overhead power lines of 110-400 kV in magnetic fields calculated to be > or = 0.01 microT. Subjects were identified by record linkages of nationwide registers. MAIN OUTCOME MEASURES: Numbers of observed cases in follow up for cancer and standardised incidence ratios for all cancers and particularly for nervous system tumours, leukaemia, and lymphoma. RESULTS: In the whole cohort 140 cases of cancer were observed (145 expected; standardised incidence ratio 0.97, 95% confidence interval 0.81 to 1.1). No statistically significant increases in all cancers and in leukaemia and lymphoma were found in children at any exposure level. A statistically significant excess of nervous system tumours was found in boys (but not in girls) who were exposed to magnetic fields of > or = 0.20 microT or cumulative exposure of > or = 0.40 microT years. CONCLUSIONS: Residentia magnetic fields of transmission power lines do not constitute a major public health problem regarding childhood cancer. The small numbers do not allow further conclusions about the risk of cancer in stronger magnetic fields. PMID- 8241852 TI - Arthroscopic surgery compared with supervised exercises in patients with rotator cuff disease (stage II impingement syndrome) AB - OBJECTIVE: To compare the effectiveness of arthroscopic surgery, a supervised exercise regimen, and placebo soft laser treatment in patients with rotator cuff disease (stage II impingement syndrome). DESIGN: Randomised clinical trial. SETTING: Hospital departments of orthopaedics and of physical medicine and rehabilitation. PATIENTS: 125 patients aged 18-66 who had had rotator cuff disease for at least three months and whose condition was resistant to treatment. INTERVENTIONS: Arthroscopic subacromial decompression performed by two experienced surgeons; exercise regimen over three to six months supervised by one experienced physiotherapist; or 12 sessions of detuned soft laser treatment over six weeks. MAIN OUTCOME MEASURES: Change in the overall Neer shoulder score (pain during previous week and blinded evaluation of function and range of movement by one clinician) after six months. RESULTS: No differences were found between the three groups in duration of sick leave and daily intake of analgesics. After six months the difference in improvement in overall Neer score between surgery and supervised exercises was 4.0 (95% confidence interval -2 to 11) and 2.0 (-1.4 to 5.4) after adjustment for sex. The condition improved significantly compared with placebo in both groups given the active treatments. Treatment costs were higher for those given surgery (720 pounds v 390 pounds). CONCLUSIONS: Surgery or a supervised exercise regimen significantly, and equally, improved rotator cuff disease compared with placebo. PMID- 8241853 TI - Cost analysis of early discharge after hip fracture. AB - OBJECTIVE: To ascertain the economic impact of an early discharge scheme for hip fracture patients. DESIGN: Population based study comparing costs of care for patients who had "hospital at home" as an option for rehabilitation and those who had no early discharge service available in their area of residence. SETTING: District hospital orthopaedic and rehabilitation wards and community hospital at home scheme. PATIENTS: 1104 consecutively admitted patients with fractured neck of femur. 24 patients from outside the district were excluded. MAIN OUTCOME MEASURES: Cost per patient episode and number of bed days spent in hospital. RESULTS: Patients with the hospital at home option spent significantly less time as inpatients (mean of 32.5 v 41.7 days; p < 0.001). Those patients who were discharged early spent a mean of 11.5 days under hospital at home care. The total direct cost to the health service was significantly less for those patients with access to early discharge than those with no early discharge option (4884 pounds v 5606 pounds; p = 0.048). CONCLUSIONS: About 40% of patients with fractured neck of femur are suitable for early discharge to a scheme such as hospital at home. The availability of such a scheme leads to lower direct costs of rehabilitative care despite higher readmission costs. These savings accrue largely from shorter stays in orthopaedic and geriatric wards. PMID- 8241854 TI - The hit and miss of ISS and TRISS. Yorkshire Trauma Audit Group. AB - OBJECTIVE: To measure interobserver variation in recording injury from case notes and its effect on calculating injury severity scores (ISS) from identical data and predicting probabilities of survival by using the combined trauma and injury severity score (TRISS). DESIGN: Observer variation study using injury severity scoring and subsequent calculation of probability of survival based on combined trauma and injury severity scores. SUBJECTS: 16 patients with a range of injury severity scores, and 15 observers. RESULTS: There was a wide variation in recorded injury severity scores, the probability of two observers agreeing on the score being 0.28 (28%). The probability of any two observers agreeing over which severity band the patient should be in was 0.5 (50%). Observer variation was independent of the training and type of observer. Survival probability (calculated by combined trauma and injury severity scoring methodology from individual observers' scores) varied by over 0.2 in six of the 16 patients and by over 0.5 in three. CONCLUSIONS: There is wide observer variation in injury severity scoring, which highlights a potential fallibility in its use for trauma audit. The use of combined trauma and injury severity scoring for individual prediction of survival is potentially inaccurate except at the extremes of probabilities. The use of the 0.5 survival line on a combined trauma and injury severity score "pre-chart" is statistically and clinically inappropriate. PMID- 8241855 TI - Care of schizophrenia in general practice. PMID- 8241856 TI - Altered inflammatory responses in smokers. PMID- 8241857 TI - Injection immunotherapy. British Society for Allergy and Clinical Immunology Working Party. AB - A working party of the British Society for Allergy and Clinical Immunology has reviewed the role of specific allergen immunotherapy in the treatment of allergic disease and produced a position statement summarising the available evidence for efficacy and safety. The working party recommends specific allergen immunotherapy for treating summer hay fever uncontrolled by conventional medication and for wasp and bee venom hypersensitivity. It is not recommended for asthma or for allergic rhinitis due to other allergens. For the recommended indications the risk:benefit ratio is acceptable provided patients are carefully selected; in particular, patients with asthma should be excluded as they are especially vulnerable to adverse reactions. Injections should be given only by doctors experiences in this form of treatment in a clinic where full resuscitative facilities are immediately available. Provided patients remain symptom free a 60 minute observation period after injection is sufficient to detect all serious adverse reactions. PMID- 8241858 TI - Acute airway obstruction after aspiration of boiling tea from teapot spout. PMID- 8241859 TI - Cost-benefit analysis. AB - Cost-benefit analysis is probably the most comprehensive method of economic evaluation available and it can be applied in two ways. The human capital approach means that the value of people's contributions is linked to what they are paid. The approach based on individuals' observed or stated preference means that their personal valuations are placed on an activity by assessing how much money they are prepared to accept for an increased risk or to pay for a particular service. Each method has its disadvantages and the most successful that has been devised so far is the "willingness to pay" (stated preference) approach, though the response to this is to a large extent dependent on the income of the person being questioned. There are still problems with its application, however, so its usefulness is limited. PMID- 8241860 TI - ABC of emergency radiology. The pelvis. PMID- 8241861 TI - Psychological distress and preterm delivery. Consider urogenital infection. PMID- 8241862 TI - Emergency dental care in east London. PMID- 8241863 TI - Fresh frozen plasma. Useful as a volume expander in children. PMID- 8241864 TI - Fresh frozen plasma. Viral inactivation may be false reassurance. PMID- 8241865 TI - Hyperbaric oxygen. Restores normal metabolic activity. PMID- 8241866 TI - Hyperbaric oxygen. Association raises awareness. PMID- 8241867 TI - Schistosomiasis. May be contracted through swimming in Lake Malawi. PMID- 8241868 TI - Epilepsy and pregnancy. Emphasise the importance of extra folate. PMID- 8241869 TI - Microalbuminuria in chronic obstructive lung disease. Consider coexistent disease. PMID- 8241870 TI - Schistosomiasis. ...contrary to popular belief. PMID- 8241871 TI - Pass rate in MRCGP examination. Depends on selection of candidates. PMID- 8241872 TI - Spinal cord stimulation and coronary ischaemia. PMID- 8241873 TI - Alcohol and bone mineral density. PMID- 8241874 TI - Diagnosis and management of systemic lupus erythematosus. Sun protection is vital. PMID- 8241875 TI - Promoting smoking cessation in general practice. PMID- 8241876 TI - Stability of vaccines. PMID- 8241878 TI - Diagnosis in histopathology. Is often unclear. PMID- 8241877 TI - Diagnosis and management of systemic lupus erythematosus. Thalidomide modifies disease. PMID- 8241879 TI - Medical management of rheumatoid arthritis. PMID- 8241880 TI - Use of diltiazem in sport. PMID- 8241881 TI - Singlehanded general practitioners and community care. PMID- 8241882 TI - Natural family planning. Effective only if used perfectly. PMID- 8241883 TI - Natural family planning. Abstinence unnatural in primates. PMID- 8241884 TI - Natural family planning. Review's enthusiasm based on flawed evidence. PMID- 8241885 TI - Babies' rectal temperature. Parents' reluctance reflects poorly on our culture. PMID- 8241886 TI - Counselling needed after screening for Down's syndrome. PMID- 8241887 TI - Fingerprints, fetal growth, and adult blood pressure. Fingerprint patterns probably inherited. PMID- 8241888 TI - Design of sharps containers. PMID- 8241889 TI - Using condoms to prevent transmission of HIV. PMID- 8241890 TI - Vibrio cholerae serogroup 0139 in England and Wales. PMID- 8241891 TI - Near patient testing. Is not cost efficient. PMID- 8241892 TI - Post tropical screening. ...unless the traveller feels unwell. PMID- 8241893 TI - Post-tropical screening. Is of little value... PMID- 8241894 TI - Ability to detect malodour might be genetic. PMID- 8241895 TI - Resuscitation in severe head injury. PMID- 8241896 TI - Tumour suppressor genes in ovarian cancer. PMID- 8241897 TI - Medical treatment at Glastonbury Festival. PMID- 8241898 TI - Drug regimens that break a country's drug laws. PMID- 8241899 TI - Funding policies for HIV and AIDS. PMID- 8241900 TI - Why are there not more women consultants? PMID- 8241901 TI - Reye's syndrome 30 years on. PMID- 8241902 TI - The need for chaperones. PMID- 8241903 TI - Melatonin. PMID- 8241904 TI - Outcomes in intensive care. PMID- 8241905 TI - The end of the GMC? PMID- 8241906 TI - Paternal radiation exposure and leukaemia in offspring: the Ontario case-control study. AB - OBJECTIVES: To test the hypothesis that there is an association between childhood leukaemia and the occupational exposure of fathers to ionising radiation before a child's conception. DESIGN: Case-control study with eight matched controls per case. SETTING: Regions of Ontario, Canada, with an operating nuclear facility. SUBJECTS: Cases were children (age 0-14) who died from or were diagnosed as having leukaemia from 1950 to 1988 and were born to mothers living in the vicinity of an operating nuclear facility. Controls were identified from birth certificates, matched by date of birth and residence at birth. There were 112 cases and 890 controls. MAIN OUTCOME MEASURES: Paternal radiation exposure was determined by a record linkage to the Canadian National Dose Registry. RESULTS: Six fathers of cases and 53 fathers of controls had had a total whole body dose > 0.0 mSv before the child's conception, resulting in an odds ratio of 0.87 (95% confidence interval 0.32 to 2.34). There was no evidence of an increased leukaemia risk in relation to any exposure period (lifetime or six months or three months before conception) or exposure type (total whole body dose, external whole body dose, or tritium dose), except for radon exposure to uranium miners, which had a large odds ratio that was not significantly different from the null value. CONCLUSIONS: The findings of this study in Ontario did not support the hypothesis that childhood leukaemia is associated with the occupational exposure of fathers to ionising radiation before the child's conception. PMID- 8241907 TI - Geographical distribution of preconceptional radiation doses to fathers employed at the Sellafield nuclear installation, West Cumbria. AB - OBJECTIVE: To examine whether the geographical distribution of births associated with preconceptional exposure of fathers to radiation at the Sellafield nuclear installation is consistent with the suggestion that this exposure explains the excess of childhood lymphoid malignancy in the adjacent village of Seascale. DESIGN: Retrospective birth cohort study. SETTING: Cumbria, West Cumbria health district, and Seascale civil parish. SUBJECTS: The 10,363 children born in Cumbria during 1950-89 to fathers employed at Sellafield. MAIN OUTCOME MEASURES: The doses of external whole body ionising radiation received by fathers at Sellafield in the total time and in the six months before conception of their children; the proportions of the collective doses associated with Seascale and the rest of West Cumbria. RESULTS: 9256 children were born to fathers who had been exposed to radiation before the child's conception. Of these, 7318 had fathers who were exposed in the six months before conception. Overall 7% (38 person-Sv) of the collective total preconceptional dose and 7% (3 person-Sv) of the collective dose for the six months before conception were associated with children born in Seascale. Of all the children whose fathers worked at Sellafield, 842 (8%) were born in Seascale. The mean individual doses before conception were consistently lower in Seascale than in the rest of West Cumbria. CONCLUSIONS: The distribution of the paternal preconceptional radiation dose is statistically incompatible with this exposure providing a causal explanation for the cluster of childhood leukaemias in Seascale. PMID- 8241908 TI - Intensive Care Society's APACHE II study in Britain and Ireland--I: Variations in case mix of adult admissions to general intensive care units and impact on outcome. AB - OBJECTIVES: To describe the extent of variation in the case mix of adult admissions to general intensive care units in Britain and Ireland and investigate the impact of such variation on outcome. DESIGN: Prospective, cohort study of consecutive admissions to intensive care units. SETTING: 26 general intensive care units in Britain and Ireland. SUBJECTS: 9099 admissions to the intensive care units studied. MAIN OUTCOME MEASURE: Death or survival at discharge before and after adjustment of case mix (age, history of chronic conditions, surgical status, diagnosis, and severity of illness) according to the APACHE II method. RESULTS: Important differences in case mix were found, with large variations between the units. Hospital mortality was significantly associated with most of the case mix factors investigated. CONCLUSIONS: Comparing crude death rates in hospital between intensive care units may be misleading indicators of performance. The collection of data on case mix needs to be standardised and differences in case mix adjusted for when comparing outcome between different intensive care units. PMID- 8241909 TI - Intensive Care Society's APACHE II study in Britain and Ireland--II: Outcome comparisons of intensive care units after adjustment for case mix by the American APACHE II method. AB - OBJECTIVES: To compare outcome between intensive care units in Britain and Ireland both before and after adjustment for case mix with the American APACHE II method and to validate the American APACHE II method in Britain and Ireland. DESIGN: Prospective, cohort study of consecutive admissions to intensive care units. SETTING: 26 general intensive care units in Britain and Ireland. SUBJECTS: 8796 admissions to the study intensive care units. MAIN OUTCOME MEASURE: Death or survival at discharge from intensive care unit and hospital. RESULTS: At discharge from both intensive care unit and hospital there was a greater than twofold variation in crude mortality between the 26 units. After adjustment for case mix, variations in mortality were still apparent. For four intensive care units the observed numbers of deaths were significantly different from the number predicted by the American APACHE II equation. The overall goodness of fit, or predictive ability, of the APACHE II equation for the British and Irish data was good, being only slightly inferior to that obtained when the equation was tested on the data from which it had been derived. When patients were grouped by various factors such as age and diagnosis, the equation did not adjust across the subgroups in a uniform manner. CONCLUSIONS: The American APACHE II equation did not fit the British and Irish data. Use of the American equation could be of advantage or disadvantage to individual intensive care units, depending on the mix of patients treated. PMID- 8241910 TI - Respiratory viruses and exacerbations of asthma in adults. AB - OBJECTIVE: To study the role of respiratory viruses in exacerbations of asthma in adults. DESIGN: Longitudinal study of 138 adults with asthma. SETTING: Leicestershire Health Authority. SUBJECTS: 48 men and 90 women 19-46 years of age with a mean duration of wheeze of 19.6 years. 75% received regular treatment with bronchodilators; 89% gave a history of eczema, hay fever, allergic rhinitis, nasal polyps, or allergies; 38% had been admitted to hospital with asthma. MAIN OUTCOME MEASURES: Symptomatic colds and asthma exacerbations; objective exacerbations of asthma with > or = 50 l/min reduction in mean peak expiratory flow rate when morning and night time readings on days 1-7 after onset of symptoms were compared with rates during an asymptomatic control period; laboratory confirmed respiratory tract infections. RESULTS: Colds were reported in 80% (223/280) of episodes with symptoms of wheeze, chest tightness, or breathlessness, and 89% (223/250) of colds were associated with asthma symptoms. 24% of 115 laboratory confirmed non-bacterial infections were associated with reductions in mean peak expiratory flow rate > or = 50 l/min through days 1-7 and 48% had mean decreases > or = 25 l/min. 44% of episodes with mean decreases in flow rate > or = 50 l/min were associated with laboratory confirmed infections. Infections with rhinoviruses, coronaviruses OC43 and 229E, influenza B, respiratory syncytial virus, parainfluenza virus, and chlamydia were all associated with objective evidence of an exacerbation of asthma. CONCLUSIONS: These findings show that asthma symptoms and reductions in peak flow are often associated with colds and respiratory viruses; respiratory virus infections commonly cause or are associated with exacerbations of asthma in adults. PMID- 8241911 TI - Use of chaperones by general practitioners. PMID- 8241912 TI - Condoms: a wider range needed. PMID- 8241913 TI - Adverse reactions to influenza vaccine in elderly people: randomised double blind placebo controlled trial. AB - OBJECTIVE: To assess the frequency and type of side effects after influenza vaccination in elderly people. DESIGN: Randomised double blind placebo controlled study. SETTING: 15 general practices in the southern Netherlands. SUBJECTS: 1806 patients aged 60 or older, of whom 904 received influenza vaccine and 902 placebo. MAIN OUTCOME MEASURES: Adverse reactions reported on postal questionnaire completed four weeks after vaccination. RESULTS: 210 (23%) patients given vaccine reported one or more adverse reactions compared with 127 (14%) given placebo. The frequency of local adverse reactions were 17.5% in the vaccine group and 7.3% in the placebo group (p < 0.001). There was no difference in systemic adverse reactions (11% v 9.4%; p = 0.34). In general, men reported fewer side effects than women. CONCLUSION: Only local side effects were more common in vaccinated patients and all side effects were mild. PMID- 8241914 TI - Who pays for community care? The same old question. AB - Trends in mental health service funding over the past 40 years show that the programme of hospital closures has not resulted in a significant release of resources to fund community based services. Far from being excessive, the current provision of residential services (both NHS and non-NHS) for mentally ill people is now below levels recommended as sufficient by the government, the Royal College of Psychiatrists, and the National Schizophrenia Fellowship. What clinical research evidence there is suggests that more rather than fewer residential places are required. This situation is likely to be compounded by the recent transfer of responsibility for funding private and voluntary residential care from the Department of Social Security to local authority social services departments. PMID- 8241915 TI - The policy context. AB - The final article in the series considers the ways in which the methods described previously are used in the formation of policy. When health authorities are making decisions about how to spend their money they have to draw on several sources of information about priorities: diktats from policy makers, opinions of consumers and of the professional bodies involved, and evidence gained from research. They must also consider the various methods of costing and select the right one for their circumstances. Some of these methods are still in the early stages of development, but more are being developed all the time and they have a valuable role in helping decision making throughout the NHS. PMID- 8241916 TI - Perfectionism in the therapeutic appraisal of methadone maintenance. PMID- 8241917 TI - Genes, environment, and addictive behavior: etiology of individual differences and extreme cases. AB - There is considerable interest in assessing the role of genetic factors in the aetiology of addiction. This paper reviews the possible contribution of a new quantitative technique, termed DF analysis after its originators DeFries & Fulker. DF analysis provides a way of examining whether aetiological factors relating to addiction as a discrete disorder (manifest as extreme scores on a diagnostic instrument) are similar to or different from those that relate to it as a continuum. For example, it is possible to estimate the contribution of familial factors to the development of a clinical condition defined in terms of an extreme score on an instrument by examining the extent of regression towards the unselected population mean of siblings' scores on that instrument. At the same time it is possible to estimate familial influences on the full range of values by examining the correlation between the scores of siblings. Insofar as the two methods produce different estimates, it is possible to infer different levels of involvement of familial factors for extreme cases than for the continuum as a whole. Information provided by such analyses could be critical when constructing models of addictive disorders and help direct more specific studies isolating particular mechanisms. PMID- 8241918 TI - Alcohol and suicide in Denmark 1911-24--experiences from a 'natural experiment'. AB - In 1916-17, prices of alcoholic beverages increased dramatically in Denmark, and alcohol consumption decreased strongly. On the basis of this 'natural experiment', the effect of variations in per capita alcohol consumption on suicide rates is estimated, and compared to Norstrom's analysis of Danish data from the period 1931-80, as well as similar analyses from other countries. It is concluded that per capita alcohol consumption is probably related to the suicide rate in Denmark, but to a less extent than in some other countries. PMID- 8241919 TI - The role of alcohol providers in prevention: an evaluation of a server intervention programme. AB - Server intervention is a relatively new approach in the attempt to reduce the incidence of drinking and driving. Although a number of evaluations have suggested that the approach may be effective, there have been few comprehensive evaluations of such programmes. The present study utilized process evaluation techniques to assess reactions to a programme developed by the Addiction Research Foundation, and a quasi-experimental design to determine the impact of the programme on the serving practices of servers. Actors portrayed behaviours often faced by servers, and observers rated the reactions of the servers, who were unaware of the simulations, to these situations. The programme appears to have been effective in changing behaviour, in that trained servers exhibited less inappropriate responses than did untrained servers. In addition the results suggested that the programme increased servers' knowledge about their obligations and potential strategies for dealing with these situations. The implications of these findings for future implementations of such programmes are discussed. PMID- 8241920 TI - Shop steward handling of alcohol-related problems. AB - Using direct observations and extensive field interviewing over a 4-year period, this paper examines practices and beliefs of shop stewards in their effects on drinking patterns and consequences. It also includes response frequencies from a random sample survey questionnaire (n = 984) that are consistent with the qualitative analysis of steward behavior. Several themes are extracted which position steward handling of alcohol-related cases as intervening between disciplinary measures of supervisors and consequences of work-related drinking of union employees. PMID- 8241921 TI - Visiting public drinking places in Oslo: an application of the theory of planned behaviour. AB - This study was undertaken to describe and obtain better insight into pub-going in the Norwegian capital applying the theory of planned behaviour (TPB). The sample comprised 1053 persons aged between 16 and 71 years who visited public drinking places in Oslo. Two thirds of the sample were men. Data were collected by means of anonymous self-administered questionnaires. The majority of the respondents reported having visited a public drinking place at least once a week during the past 12 months. In the applied model, intention to visit a public drinking place during the next 14 days is considered to be a joint function of the attitude towards pub-going (Aact), subjective norms (SN), and perceived control over the behaviour (PBC). The goodness of fit (R2) of the model was 22%. The relative importance of the model's components were in descending order (beta): PBC, Aact and SN. Previous behaviour was included as a predictor in the TPB, and had a stronger effect upon intention than the model's original components. In conclusion, the TPB provided an adequate understanding of why pub and cafe guests frequent public drinking places. However, it is possible that the model over simplifies the processes connected to pub-going and fails to describe the dynamics of the relationship between the individual and the environment. PMID- 8241922 TI - Initial and current drug use: how are they related? AB - The present study examines how initial and current patterns of drug use are related among a group of drug users in Edinburgh. The study categorised individuals according to the year in which they first used illicit drugs. 1984 has been shown to be the year when patterns of drug use in this group began to change after high levels of heroin use in the late 1970s and early 1980s. Thus 1984 was used as the intercept point to create two groups: the epidemic group, comprising individuals who began using illicit drugs before 1984, and, the post epidemic group, who began using during or after 1984. Data from individuals were gathered from interviews and medical records. Statistical analysis revealed interesting differences between the groups, for example, the groups did not differ significantly in their use of methadone but the epidemic group used more methadone obtained on prescription. The post epidemic group was found to inject more often, but, did not share injecting equipment more than the epidemic drug users. A greater number of the epidemic group began using drugs by injecting and a greater number of the post epidemic group began by taking drugs orally. The issues raised by the results are discussed, and, it is suggested that the characteristics of the drug using community within which an individual begins using drugs may exert influence on patterns of drug use in later years. PMID- 8241923 TI - Does post-withdrawal cue exposure improve outcome in opiate addiction? A controlled trial. AB - A controlled trial studied whether cue exposure prevented relapse in opiate addiction. Subjects were randomly allocated to one of two inpatient treatment settings: a drug dependence unit with a special 10 week program and 4 weeks in a behavioural/general treatment unit without such a program. In each setting, following drug-withdrawal, subjects had either cue exposure for at least six sessions over 3 weeks, or a control condition. Subjects were followed up twice, at about 6 weeks and 6 months post-treatment. 186 subjects were randomly allocated; 69 were assessed post-detoxification, and of these 43 completed cue exposure or control treatments. Cue exposure and control subjects did not differ in cue reactivity. This was evaluated post-treatment for cue exposure subjects and at a comparable time point for controls. All groups showed a significant decrement in cue-elicited craving, withdrawal responses and negative mood. Cue exposure and control subjects did not differ at either of the two follow up interviews. PMID- 8241924 TI - Opiate and cocaine consumers attending Barcelona emergency rooms: a one year survey (1989). AB - Due to the limitations of standard epidemiological methods, indirect indicators have often been used to describe the characteristics of drug abusing populations and to assess prevalence trends in illegal drug use. In Barcelona (Spain), a study of emergency room (ER) attendance was carried out to describe the population of opiate/cocaine consumers across the whole city who use this service. Three thousand four hundred and five consumers of opiates and/or cocaine, aged 15-44 years, who attended ERs during 1989, were identified. They accounted for 6807 episodes in the hospitals surveyed. Their mean age was 26 years, men (73%) being 1 year older than women (25.2 years). The drug of abuse was specified in the clinical records of 60% of individuals, heroin being the most frequently specified (56%). The main reason for attendance was 'other medical condition' (OMC) (55% of episodes), followed by withdrawal (34%) and overdoses (6%). Seventy-one percent of individuals were residents of Barcelona city, yielding a rate of 3.2 opiate/cocaine consumers attending ERs per thousand Barcelona residents aged 15-44. The geographical distribution of the rates in the city showed a very large difference between districts, the most deprived ones having a higher rate of consumers attending ERs. ER data can provide valuable insights into the nature and dimensions of drug abuse problems. PMID- 8241925 TI - AIDS risk behaviors and knowledge among heterosexual alcoholics and non-injecting drug users. AB - The disinhibitory effects of abusable substances on sexual behavior and the increasing HIV prevalence among heterosexuals suggest that alcoholics and non injection drug users may be at risk for HIV infection. We examined alcohol and non-injection drug use as AIDS risk factors, AIDS risk knowledge, and the effect of AIDS education upon voluntary HIV testing among 91 heterosexual male inpatients in a VA alcohol rehabilitation program. Questionnaire data revealed relationships between age, the use of alcohol, marijuana and intranasal cocaine just prior to sex and an increase in the number of female sexual partners. Use of alcohol just prior to sex was also associated with an increased number of unprotected sexual behaviors. AIDS risk knowledge in our sample was comparable to norms from previous studies. Inpatients received education concerning alcohol and sexuality either with or without an AIDS component. AIDS education and offer of HIV testing were associated with increased requests for HIV testing. PMID- 8241926 TI - The Significant-other Behavior Questionnaire (SBQ): an instrument for measuring the behavior of significant others towards a person's drinking and abstinence. AB - The psychometric properties of the Significant-other Behavior Questionnaire (SBQ), an instrument measuring the specific social support in terms of the patient's drinking. Assessments of the significant other's behavior in the presence of the patient drinking is provided by both the patient and significant other. Data from the two SBQ forms were analyzed with a principal axis factor analytic procedure with varimax rotation. The scales were similar for the two forms with four behavioral reactions, withdraw from the patient, support drinking, support sobriety, and punish drinking. Correlations between the two forms on the same four scales were higher than correlations within groups on three of four scales and differed from measures of general social support. Applications of the SBQ are discussed. PMID- 8241927 TI - The families of problem drug users: a study of 50 close relatives. AB - The research described in this paper resulted from a collaborative multi-centre study of the relatives of problem drug users involving six practitioners and researchers, in four centres within the south-west of England. The objective of this research was simply to interview 50 close relatives of identified problem drug users, with the identification occurring through clinics and self-help groups. Quantitative and qualitative results show that large numbers of these relatives reported many negative experiences. The partners of illicit drug users reported both more and differently patterned problematic behaviours than those of prescribed tranquillizer users. The partners of illicit users also reported different problems to those of the parents. Relatives reported many negative effects in terms of how they viewed the drug user, and how the experiences had affected their health. They also described various coping mechanisms, and the extent of the support which they had received. The results are discussed in terms of coping, and similarity with research into the families of problem drinkers. PMID- 8241928 TI - Hair analysis for monitoring drug use. PMID- 8241929 TI - Further ethical issues raised by hair analysis for drugs. PMID- 8241930 TI - Routine peroperative cholangiography. PMID- 8241931 TI - The quest for a herbal contraceptive. PMID- 8241932 TI - Chronic hepatitis in a large Indian hospital. AB - BACKGROUND: In developed countries as well as in Southeast Asia, the hepatitis B and C viruses are the main causes of chronic hepatitis. In India, however, there have been no major investigations on the aetiology of chronic hepatitis. (The hepatitis E virus which is responsible for half the sporadic and most of the epidemic cases of acute viral hepatitis in India does not cause chronic disease.) We, therefore, studied the profile of chronic hepatitis in India. METHODS: The clinical presentation, aetiology, serology and histological changes were studied prospectively in 48 patients with chronic hepatitis admitted to the All India Institute of Medical Sciences, New Delhi. Of these, 44 (92%) had chronic active hepatitis, 3 (6.3%) had chronic persistent hepatitis and 1 (2%) had chronic lobular hepatitis. RESULTS: The hepatitis B virus was the aetiological agent in 24 (50%) of these patients, the hepatitis D virus in association with hepatitis B virus in 10 (21%), the hepatitis C virus in 7 (15%) and the non-A, non-B viruses other than the hepatitis C virus in 6 (13%). One patient (2.0%) had autoimmune chronic active hepatitis. Jaundice at presentation was seen in 33 (69%) patients and more than half had hypoalbuminaemia (< 3 g/dl) with a prolonged prothrombin time. Alanine aminotransferase levels were less than 5 times above normal in over two-thirds of the patients. The highest alanine aminotransferase values were observed in patients with hepatitis D virus infection whereas the lowest were seen in patients with non-A, non-B related chronic active hepatitis. Histological examination revealed bridging necrosis in 40 (91%) patients with chronic active hepatitis indicating a severe form of disease. Replication of the hepatitis B virus was seen in 13 patients with chronic hepatitis, 5 of whom had hepatitis D virus-induced chronic hepatitis. Patients with hepatitis B virus replication had higher alanine aminotransferase values and more severe bridging necrosis than patients who did not have replicating viruses. Higher alanine aminotransferase values, ascites and oesophageal varices were encountered more frequently in patients with hepatitis B and D virus than in those with non-A, non-B related chronic hepatitis. CONCLUSION: Chronic hepatitis is not uncommon in India. It presents with evidence of severe disease and, as elsewhere, is most frequently caused by the hepatitis B virus. PMID- 8241933 TI - Filariasis and blood groups. AB - BACKGROUND: Using modern statistical techniques, we investigated the controversial relationship between lymphatic filariasis and blood groups in a very large number of patients. METHODS: The ABO blood group, microfilaraemia and symptomatic disease status of 1444 persons was determined. The association between the filarial status and ABO blood group was studied by the Chi-square test and the method of logistic regression by fitting a model. This method simultaneously tested for association of factors such as blood groups and age with the occurrence of parasitaemia or filarial disease. RESULTS: The ABO blood groups were not risk factors in the occurrence of patent parasitaemia or filarial disease though age was associated with the occurrence of microfilaraemia and symptomatic disease. CONCLUSION: ABO blood groups are not associated with the occurrence of bancroftian filariasis. PMID- 8241934 TI - Short stay thyroidectomy in a tertiary care hospital. AB - BACKGROUND: Many procedures which earlier required a prolonged stay in hospital are now being performed on a short stay basis. METHODS: In 1989 we adopted a policy of 'short stay thyroidectomy' defined as a postoperative stay of less than 48 hours. Till 1992, 162 patients were included in this study. After early drain removal at 16-43 hours (median 38), these patients were discharged 18-45 hours (median 41) after surgery. We recorded the postoperative drainage of 155 of these patients. RESULTS: Out of 162 patients, 156 were discharged within 48 hours after surgery. No patient developed any complication which required re-admission and there was a fall in the amount of drainage within 12-18 hours. CONCLUSION: Short stay thyroidectomy is feasible in a developing country. PMID- 8241935 TI - Endoscopic management of postoperative bile leakage. PMID- 8241936 TI - Percutaneous nephrostomy in malignant urinary tract obstruction. PMID- 8241937 TI - Long term domiciliary oxygen therapy in chronic obstructive pulmonary disease. PMID- 8241938 TI - Oral ulcers. PMID- 8241939 TI - A young female with a stroke and limb gangrene. PMID- 8241940 TI - The National Institute of Nutrition, Hyderabad. PMID- 8241941 TI - Health care fraud. PMID- 8241942 TI - Academic staff in our teaching hospitals. PMID- 8241943 TI - On organ transplantation. PMID- 8241944 TI - On organ transplantation. PMID- 8241945 TI - Continuing medical education for general practitioners. PMID- 8241946 TI - On organ transplantation. PMID- 8241947 TI - Recombinant human erythropoietin for the treatment of chronic anemia in multiple myeloma and squamous cell carcinoma. AB - Recombinant human erythropoietin (rHuEPO) improves chronic anemia of cancer, but the proportion of patients who respond favorably to the treatment varies depending on the type of neoplasia. Preliminary data of the two malignancies with the highest response rates, namely, multiple myeloma and squamous cell carcinoma, are reported. Twenty patients with multiple myeloma and 14 with squamous cell carcinoma, who had presented with hemoglobin levels < 11 g/dl, were treated with rHuEPO, 150 U/kg, three times/week. Response, defined as an increase of at least 2 g/dl hemoglobin within 12 weeks, was achieved by 15 myeloma patients (75%) and 11 patients with squamous cell carcinoma (79%). Tolerance of the treatment was excellent. The WHO performance status and quality of life improved in responders. The remarkably low levels of endogenous EPO in our patients with squamous cell carcinoma, most of whom had been treated with cisplatin-or carboplatin-containing regimens, suggest that anemia in these cases had been at least partly chemotherapy induced. In myeloma patients, the blunted EPO response to the anemic condition may have been partly caused by subclinical tubular insufficiency induced by toxic paraproteins. Future studies should aim to elucidate factors which are responsible for the inability of some patients to respond to rHuEPO treatment, even though in multiple myeloma and squamous cell carcinoma these non responders are a small minority. PMID- 8241948 TI - Stringently purified human hematopoietic progenitors/stem cells: analysis of cellular/molecular mechanisms underlying early hematopoiesis. AB - Analysis of the cellular/molecular basis of the early steps of hematopoietic proliferation and differentiation is hindered by the rarity of hematopoietic progenitors and stem cells (HP/HSC). The intensive efforts devoted to the development of purification methods for early HP and HSC, although initially largely unsuccessful, have recently provided a high level of HP/HSC yield and/or recovery. The methodology developed by our group, recently improved, provides not only virtually complete purification, but also abundant recovery of early HP/HSC such as colony forming units granulocyte/erythroid/macrophage/megakaryocyte (CFU GEMM), burst forming units erythroid (BFU-E), CFU granulocyte/macrophage (CFU GM)/CFU blast cells (CFU-B), and long-term culture initiating cells (LTC-IC) from adult peripheral and cord blood (CB). We have also developed a serum-free liquid suspension culture for unilineage erythroid (E), granulocytic (G) or monocytic (M) differentiation of stringently purified HP/HSC. These culture systems allow sequential collection and cellular/molecular analysis of discrete populations of hematopoietic cells at a homogenous stage of differentiation specifically along a unilineage pathway. These experimental tools have been utilized to investigate cellular/molecular mechanisms underlying early hematopoiesis. The transcription factor (TF) GATA-1 is considered to be the "master" gene of erythropoiesis. In highly purified HP/HSC undergoing E or GM differentiation, GATA-1 expression is characterized initially by proliferation-dependent activation and at later stages by sustained expression in the E pathway and suppression in the GM pathway. Hypothetically, similar on/off switches of lineage-restricted TF may underlie the binary fate decisions of early HP differentiation. The expression and modulation of hematopoietic growth factor receptors (HGFR) in early hematopoiesis have been extensively analyzed. The results suggest a model of transactivation cascade for HGFR such as interleukin 6 receptor (IL-6R), IL-3R, GM colony stimulating factor receptor (GM-CSFR), and erythropoietin receptor (EpR), whereby each HGF upmodulates the R(s) for distal-acting HGF(s). Finally, we have investigated the effect of HGF on reactivation of hemoglobin F (HbF) in clonogenic or liquid suspension serum-free culture of purified adult HP. The results suggest that c kit ligand (KL) plays a key role in the reactivation of HbF synthesis in adult life, and IL-3/GM-CSF potentiate this effect at low KL level. The KL-induced HbF reactivation is seemingly related to an enhanced proliferation of early E progenitors in their differentiation pathway. PMID- 8241949 TI - Signal transduction in the erythropoietin receptor system. AB - Developing erythroid cells require the glycoprotein hormone, erythropoietin (EPO) as an activator of the rapid proliferation of early proerythroblasts (colony forming units-erythroid [CFU-e]), and subsequently as an activator of late erythroid gene expression. Activation of these growth and differentiation events proceeds from the binding of EPO at its transmembrane receptor (Class I cytokine receptor), to the engagement of a complex set of signaling pathways. Studies of reconstituted activities of the cloned EPO receptor in transfected hematopoietic cell lines have served well in identifying receptor domains and downstream mediators involved in proliferative signaling. Extracellular domains have been defined which contribute to ligand binding, receptor processing and transport, and possible dimerization. Cytosolic regions have been delineated which mediate induced mitogenesis, early gene transcription, activated protein tyrosine phosphorylation, down modulation of EPO- and granulocyte-macrophage colony stimulating factor (GM-CSF)-induced proliferation, and direct association with PI3- and JAK-2 kinases. These newly defined properties begin to align the EPO receptor mechanistically with growth factor receptors (GFR) which encode, or likewise associate with, regulated protein tyrosine kinases including the Class II cytokine receptors for interferons alpha/beta and gamma. An improved understanding of factors which mediate EPO-induced late erythroid gene activation also is emerging. These factors and pathways may be distinct from those associated with EPO-induced proliferation and may involve induced increases in cellular Ca++, cAMP and arachidonic acid, as well as the modulation of GATA-1, and/or SCL. Attributes of model systems used in studies of the role of EPO in late erythroid differentiation also are considered. PMID- 8241950 TI - Partial amelioration of AZT-induced macrocytic anemia in the mouse by folic acid. AB - CBA/Ca mice being maintained on azidothymidine (AZT) in drinking water were given vitamin B12 and folate in an effort to ameliorate the macrocytic anemia associated with AZT administration. The B12/folate regimen was ineffectual, but higher doses of folate given daily resulted in an increase in RBC and a decrease in mean corpuscular hemoglobin (MCH) and polychromatophilic erythrocytes (PCE) while mean corpuscular volume (MCV) remained relatively constant. The implications of these findings on RBC production and hemoglobin synthesis are discussed. PMID- 8241951 TI - Effects of macrophage colony stimulating factor and granulocyte-macrophage colony stimulating factor on osteoclastic differentiation of hematopoietic progenitor cells. AB - Although the hematopoietic origin of the osteoclast is generally accepted, the precise phenotype of the progenitor and the regulation of its differentiation are unclear. This study compares proliferation and differentiation of progenitors in response to macrophage colony stimulating factor (M-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF). Nonadherent progenitor cells from murine long-term bone marrow cultures (LTBMC) (as a source of osteoclast progenitors) demonstrated a significant proliferative response to M-CSF. In addition, M-CSF increased the number of multinucleated cells, only a small percent of which (14-16%) were tartrate-resistant, acid phosphatase (TRAP) positive. In contrast, cells cultured with GM-CSF generated more TRAP-positive multinucleated cells even at concentrations less stimulatory of proliferation than M-CSF. The osteoclast phenotype of these multinucleated cells was also assessed by ultrastructural characterization of ruffled borders in association with bone fragments. The bone-active hormone 1,25-dihydroxyvitamin D3 inhibited the proliferation of this subset of progenitor cells in the presence of M-CSF or GM-CSF. All of these results show effects on progenitors in the absence of the stromal cell microenvironment in this system. These results provide evidence for a divergence in the biological responsiveness of osteoclast progenitor cells to M CSF compared with GM-CSF; they support the notion that M-CSF has a "priming" effect on osteoclast progenitors whose subsequent differentiation to osteoclastic multinucleated cells is promoted by GM-CSF. PMID- 8241952 TI - Antitumor effect of interleukin 7 in combination with local hyperthermia in mice bearing B16a melanoma cells. AB - Interleukin (IL)-7 has been evaluated for its influence, alone or in combination with local hyperthermia (LH), on B16a melanoma-bearing mice. Six- to eight-week old C57BL/6J male mice were inoculated s.c. with 5 x 10(5) tumor cells into the left hind limb. Mice were randomly divided into four groups, and treated s.c. with IL-7 (5 ng) or saline as control, twice a day for three weeks beginning eight days after tumor inoculation. LH, using hot water circulator at 43 +/- 0.2 degrees C for 30 min, was induced to the limb with tumor twice a week for two weeks. Size of the primary tumor was measured every other day for five weeks. Mice were sacrificed five weeks after tumor inoculation. The size of the primary tumor and the number of lung metastases were reduced in mice treated either with IL-7 or LH alone. As a control for IL-7, granulocyte colony stimulating factor (G CSF) alone had no effect on primary tumor size or number of lung metastases. The greatest antitumor effect was observed in mice treated with IL-7 in combination with LH. Survival was prolonged significantly only in mice treated with IL-7 plus LH compared with that of mice treated with saline. Decreased natural killer (NK) cell activity, number of Thy1.2 cells, and ratio of L3T4+/Lyt2+ cells were associated with tumor growth. These parameters were restored in mice treated with IL-7 plus LH. Increases in levels of IL-1 alpha, IL-6, tumor necrosis factor (TNF alpha) and interferon (IFN gamma) were associated with an increase in the survival of tumor-bearing mice treated with IL-7 and/or LH. These results suggest that changes in T-cell, NK cell and cytokines such as IL-1 alpha, IL-6, TNF-alpha and IFN-gamma in response to IL7 and/or LH might account for prolonged survival of B16a melanoma-bearing mice and that IL-7 might be useful as a potential antitumor agent combined with other therapy in certain malignant solid tumors with metastases. PMID- 8241953 TI - The molecular specificity of action of the tetrapeptide acetyl-N-Ser-Asp-Lys-Pro (AcSDKP) in the control of hematopoietic stem cell proliferation. AB - The hemoregulatory tetrapeptide Acetyl-N-Ser-Asp-Lys-Pro (AcSDKP, M(r) = 487 amu) prevents the recruitment of hematopoietic stem cells into S phase. AcSDKP does not possess inherent inhibitory activity against hematopoietic stem cells, rather it appears to act by "blocking" the action of a hematopoietic stem cell proliferation stimulator present in extracts of regenerating hematopoietic tissue. The molecular specificity of this action was investigated using a number of peptide analogues of AcSDKP and evaluating their ability to "block" the recruitment of a primitive murine hematopoietic precursor (high proliferative potential colony forming cell HPP-CFC) into S phase following incubation with a hematopoietic stem cell proliferation stimulator. The capacity of AcSDKP to compromise the action of the hematopoietic stem cell proliferation stimulator was not evident for either AcSDDKP or AcSD beta KP, two structurally distinct forms of the molecule; similar no stimulator-"blocking" activity was observed for the tripeptide Ala-Asp-Lys (ADK, M(r) = 332 amu), while the tripeptide Ser-Asp-Lys (SDK, M(r) = 348 amu) was active. It would appear that the hemoregulatory action of AcSDKP is specific and that the tripeptide sequence SDK may be a significant component of the molecule. PMID- 8241954 TI - Burst-promoting activity in the serum of patients with chronic renal failure undergoing long-term hemodialysis. AB - Anemia in patients with chronic renal failure (CRF) is multifactorial, and while the majority will respond to the paternal administration of recombinant human erythropoietin (r-HuEPO), a role for coexistent plasma inhibitors and stimulators, such as burst-promoting activity (BPA), remains controversial. To evaluate the latter possibility, eight individuals with CRF on long-term hemodialysis, were studied before (mean hemoglobin 58.4 +/- 8.0 g/l and after 12 weeks of r-HuEPO therapy (mean hemoglobin 100.4 +/- 18.3 g/l). In vitro erythroid cultures using erythroid burst forming unit (BFU-E) and erythroid colony forming unit (CFU-E) assays were performed, plating 5 x 10(4) bone marrow mononuclear cells and comparing growth in heat-inactivated autologous serum with AB serum. Using Step III sheep erythropoietin (Connaught Laboratories, Willowdale, Ontario, Canada) (n = 4), mean BFU-E pre-therapy were 89.7 +/- 75.1, CFU-E were 418.5 +/- 150.6, whereas the corresponding figures in AB serum were 2.5 +/- 2.9 and 197 +/- 94.19, p = 0.1, p = 0.01, respectively. Similarly, with r-HuEPO (EPOCONN: Connaught Laboratories, Willowdale, Ontario, Canada) (n = 4), mean BFU-E were 145.25 +/- 103.3 in autologous serum and 31.0 +/- 56.75 in AB serum (p = 0.04). As controls, erythroid progenitors from two normal donors yielded 69 and 61 BFU-E colonies in autologous serum and 103 and 42 in AB serum; the corresponding CFU-E were 52 and 235 versus 136 and 137.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241955 TI - Mobilization of peripheral blood stem cells by subcutaneous injections of yeast derived granulocyte macrophage colony stimulating factor: a phase I-II study. AB - A phase I-II study was conducted in 15 patients to determine the tolerability, efficacy and optimal dose of s.c. yeast-derived granulocyte-macrophage colony stimulating factor (GM-CSF) for mobilization of peripheral blood stem cells (PBSC). PBSC were measured by the colony forming unit granulocyte-macrophage (CFU GM) and burst forming unit erythroid (BFU-E) in vitro colony assays and CD34 flow cytometric analysis. GM-CSF was administered for 12 days as a single daily s.c. injection at 125 micrograms/M2 (n = 3), 250 micrograms/M2 (n = 3), 375 micrograms/M2 (n = 6) and 500 micrograms/M2 (n = 3). At these doses, sustained serum GM-CSF levels of approximately 100-400 pg/ml were observed for at least six hours. Most (97.2% or 175/180) of the planned GM-CSF doses were administered with no serious adverse effects at any of the dose levels. Three leukapheresis procedures were performed before and then after 8, 10 and 12 doses of s.c. GM CSF. Compared with basal leukapheresis collections, s.c. GM-CSF mobilized leukapheresis collections yielded greater numbers of total white blood cells ([WBC] 15/15 patients or 100%, p < 0.001) mononuclear cells ([MNC] 8/15 patients or 53%, p = 0.55), CFU-GM (11/15 patients or 73%, p = 0.037), BFU-E (9/15 patients or 60%, p = 0.13) and CD34+ cells (6/15 patients or 40%, p = 0.15). The in vitro CFU-GM were augmented approximately two to tenfold in GM-CSF mobilized collections with no clear dose effects from 125-500/micrograms/M2. However, in vivo engraftment time to absolute neutrophil count (ANC) 500/mm3 correlated strongly with GM-CSF PBSC mobilizing dose (r2 = 0.99). We conclude that s.c. yeast-derived GM-CSF is a simple, effective and tolerable method for mobilization of PBSC. PMID- 8241956 TI - The tetrapeptide AcSerAspLysPro (Seraspenide), a hematopoietic inhibitor, may reduce the in vitro toxicity of 3'-azido-3'-deoxythymidine to human hematopoietic progenitors. AB - 3'-azido-3'-deoxythymidine (AZT), the main antiviral drug used in AIDS treatment, is known to induce anemia and neutropenia. These effects have been attributed to its toxicity to hematopoietic progenitors. In this report, we present a new approach to reduce AZT hematotoxicity by using an inhibitory factor of the hematopoietic stem cells, the tetrapeptide AcSerAspLysPro (AcSDKP, Seraspenide), which has been shown to increase the survival of mice subjected to high doses of chemotherapy and to block reversibly the cycling of human granulocyte-macrophage colony forming unit (CFU-GM) and burst forming unit erythroid (BFU-E) progenitors. Normal bone marrow mononuclear cells (BMMNC) from 14 subjects were incubated with or without AcSDKP (10(-10) M) for 20 h and with or without AZT (100 microM) for another 2 h. After washing, cells were plated in methylcellulose in the presence of interleukin 3 (IL-3), granulocyte-macrophage colony stimulating factor (GM-CSF) and erythropoietin (EPO). Under these conditions, the preincubation of cells with AcSDKP reduced significantly the toxicity of AZT to both BFU-E and CFU-GM at least in 3 out of 8 and 4 out of 10 cases, respectively. A careful statistical analysis of these observations indicates that AcSDKP may be an efficient factor in preserving progenitors against AZT-induced hematopoietic toxicity. PMID- 8241957 TI - Immunolabelling of the presynaptic membrane of Torpedo electric organ nerve terminals with an antiserum towards the acetylcholine releasing protein mediatophore. AB - Mediatophore is a nerve terminal membrane protein purified from Torpedo electric organ on its ability to translocate acetylcholine upon calcium action. An antiserum able to immunoprecipitate mediatophore activity was used to study the subcellular distribution of this protein. The presynaptic membrane exhibited a strong and discontinuous immunogold labelling, especially at the active zone where ACh is thought to be released. Two antigens were recognized on immunoblots of synaptosomal membranes: the 15-kDa subunit of mediatophore and a 14-kDa membrane protein that has a wide non-neuronal distribution. Antibodies purified from the serum on native mediatophore and monospecific towards the 15-kDa antigen still gave a high presynaptic membrane localized labelling. In addition, a few 14 kDa protein sites were present at the active zone. The Schwann cell finger interposed between the presynaptic membrane and the postsynaptic arch also exhibited the 14-kDa antigen raising the question of a possible interaction of mediatophore with the 14-kDa protein originating from the Schwann cell. PMID- 8241958 TI - Structural analysis of glycosyl-phosphatidylinositol membrane anchor of the Toxoplasma gondii tachyzoite surface glycoprotein gp23. AB - In this study we describe the biochemical features of the Toxoplasma gondii tachyzoite surface glycoprotein, gp23, demonstrating that it is attached to the parasite membrane by a glycosyl-phosphatidyl inositol anchor. Gp23 was metabolically labeled with tritiated palmitate, myristate, ethanolamine, inositol, glucosamine, mannose and galactose, as expected for a GPI-anchor structure. Gp23 was released from the surface of living parasites after treatment with phosphatidylinositol-specific phospholipase C(PI-PLC) and the resulting water-soluble protein was immunoprecipitated with a monoclonal antibody specific for gp23. The GPI-core glycan was generated after aqueous-HF dephosphorylation followed by nitrous acid deamination and its carbohydrate structure was analyzed using selective exo- and endoglycosidase treatments. Finally, the phosphatidylinositol moiety of gp23 was characterized using PI-PLC and phospholipase A2 (PLA2) digestions. Our cumulative data suggest that gp23 of T gondii tachyzoites contains a modified GPI-backbone similar to the mammalian Thy 1 anchor, consisting of a conserved core structure (ethanolamine-PO4-6-Man alpha1 2-Man alpha1-6-Man alpha1-4-GlcN alpha1-6-PI) bearing beta-linked N acetylgalactosamine residue(s). PMID- 8241959 TI - In vitro differentiation and mineralization of cartilaginous nodules from enzymatically released rat nasal cartilage cells. AB - Nasal cartilage cells from 21-day-old rat fetuses were cultured at high density in the presence of ascorbic acid and beta-glycerophosphate over a 12-day period. Immediately after plating, the cells exhibited a fibroblastic morphology, lost their chondrocyte phenotype and expressed type I collagen. On day 3, clusters of enlarged polygonal cells were found. These cell clusters synthetized type II collagen and formed an alcian-blue-positive matrix. The following days, a progressive increase in the number of cells positive for type II collagen was noted and, on day 8, typical cartilaginous nodules were formed. These nodules increased in size and number, spreading outward, laying down a dense matrix which mineralized. Light and electron microscopy observations of cross-sections of nodules confirmed the cartilaginous nature of this tissue formed in vitro with typical chondrocytes embedded in a hyaline matrix. Furthermore, at the electron microscopic level, matrix vesicles were seen in extracellular matrix associated with the initiation of mineralization. Typical rod-like crystals were present in the intercellular spaces along the collagen fibers. These results indicated that in a specific environment, dedifferentiated chondrocytes were able to redifferentiate and to form nodular structures with morphological ultrastructure of calcified cartilage observed in vivo. PMID- 8241960 TI - Triiodothyronine influences quail myoblast proliferation and differentiation. AB - The influence of triiodothyronine (T3) on avian myoblast proliferation and differentiation was studied in secondary cultures using plating densities of 2,500 and 7,000 cells/cm2. Culture media were depleted of T3 (control myoblasts) and increasing amounts were then added to concentrations of 0.6, 3 and 15 nM T3 (treated myoblasts). Independent of the cell density, T3 induced a dose-related decrease in myoblast proliferation measured by cell number, doubling time and 3H thymidine incorporation. However, with the lower plating density, this influence was delayed, occurring only after the third day of culture for 0.6 nM T3-treated myoblasts and simultaneous with the onset of myosin heavy chain accumulation. Moreover, when myoblasts were exposed to BrdU for 48 h, the T3 growth inhibitory effect disappeared, thus showing that this effect was clearly linked to differentiation. In addition, we have shown that T3 induced an early fusion of myoblasts: 65% of the maximal value of the fusion index was reached on day 3 in the T3-treated cells in comparison to 25% in the control myoblasts. This hormone also enhanced accumulation of muscle-specific proteins (connectin, acetylcholine receptors, myosin heavy chain), tested by cytoimmunofluorescence, ELISA, binding experiments and Western blot. All these results show that T3 increased myoblast differentiation through a pathway including myoblast withdrawal from the cell cycle. The influence of T3 could partly explain its previously reported positive effect on the number of muscle fibers. PMID- 8241961 TI - Endocytic activity of male germ cells during puberty. AB - It is well known that numerous germ cells degenerate during the first meiotic division and spermatid elongation is due mostly to an adverse physiological microenvironment in relation to hormonal deficiency. The present study is aimed at investigating the endocytic activity of germ cells, during the first wave of mouse spermatogenesis, using transferrin coupled to gold particles, in order to study the efficiency of this possible pathway of communication between Sertoli cells and germ cells. Labelling experiments in control animals confirmed a receptor-mediated pathway in all germ cells during puberty. Furthermore, our morphological and quantitative data revealed that during spermatid elongation, degenerating germ cells possessed a highly developed endocytic apparatus which contained twice as many transferrin gold particles than normal adult cells. The fact that endocytosis of transferrin was increased in degenerating germ cells indicates that, most probably, germ cell degeneration during the first wave of spermatogenesis did not result from a deficiency in iron transport. The higher endocytic activity of degenerating germ cells, compared to adult control cells, could not only be the result of a simple process of plasma membrane internalization but also a complex mechanism which could be involved in the degradation of the cells. PMID- 8241962 TI - Localization of a Band 3-related protein in the mitochondria-rich cells of amphibian skin epithelium. AB - Based on immunoblotting procedure, the isolated epithelium of amphibian skin was found to contain a 180 kDa protein which cross-reacts with a polyclonal antiserum raised against human erythrocyte Band 3. Immunoperoxidase and immunofluorescence staining techniques indicated that the Band 3-related protein was localized in the mitochondria-rich cells (MRC) of this epithelium, with characteristic apical labelling pattern. Our findings show that the putative apical anion exchanger of the MRC is immunologically related to the band 3 multigenic family, which catalyzes Cl(-)-HCO3- transmembranous exchange. It thus suggests a molecular basis for the role played by these cells in the transepithelial Cl- pathway and acid-base regulation. PMID- 8241963 TI - The cortical actin cytoskeleton in a Dipteran embryo: analysis of the spatial reorganization of F-actin aggregates during the early nuclear division cycles. AB - Rhodamine phalloidin-staining was used to study the organization of the cortical actin cytoskeleton of the early Ceratitis capitata embryo. The dynamics of the actin aggregates and their changes in distribution during the formation of the syncytial blastoderm, were followed in detail. It was found that these aggregates formed a shell-like cluster around the interphase nuclei, and concentrated toward the poles of the mitotic apparatus when the nuclei divided. Laser scanning confocal microscopy revealed that aggregates not clustered at the poles of the mitotic apparatus were closely associated with fine fibers of a dense cytoplasmic network of actin filaments. PMID- 8241964 TI - African swine fever virus interaction with microtubules. AB - The role of microtubules in intracellular transport of African swine fever virus (ASFV) and virus-induced inclusions was studied by immunofluorescence using anti ASFV and anti-tubulin antibodies, by electron microscopy of infected Vero cells and by in vitro binding of virions to purified microtubules. MTC, a reversible colchicine analogue, was used to depolymerize microtubules. In cells treated with MTC multiple large inclusions containing ASFV antigens and particles were observed in the cytoplasm. Removal of the drug lead to migration and fusion of the inclusions at a perinuclear location. To study the effect of microtubule repolymerization on virus particle distribution, the particles were counted in thin sections of MTC treated cells and at different times after removal of the drug. In cells treated with MTC 6.8% and 3.6% of the virus particles were found respectively in the cytoplasm and at the cell membrane while 38% of the particles were located around the virosome. With reversal of the drug effect the number of virus particles around the virosomes progressively decreased to 10% at 2 h while the number of particles in the cytoplasm and at the cell membrane increased. At 2 h after removal of the drug 33.5% of the particles were found budding from the cell membrane. Virus particles were found closely associated with microtubules in cytoskeletons obtained by Triton X-100 extraction of taxol treated cells. The association of virus particles with microtubules was also observed in vitro using purified microtubules and virus particles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241965 TI - The impact of hearing impairment on television viewing in the UK. AB - Just under one in 10 of a nationally representative sample of UK television viewers said that they experienced difficulty with their hearing. These hard-of hearing viewers were found to report much greater difficulty watching programmes (mean difficulty rating = 32%) than elderly viewers with no reported hearing difficulty (mean difficulty rating = 10%), or those viewers generally who said they had no hearing problems (mean difficulty rating = 3%). Using a similarly constructed rating for reported enjoyment of different television programmes, hard-of-hearing viewers were found to exhibit a small reduction in enjoyment across the majority of programmes types. While it might have been anticipated that a greater proportion of those with impaired hearing owned a teletext television set giving them access to subtitling, this was not found to be the case. Across the sample as a whole, teletext ownership was shown to be 45%, but was lower than this (38.5%) amongst the hard-of-hearing. The findings corroborate what has been shown in another study, namely that teletext ownership is lower among older viewers. Hearing impairment, if it is not congenital or of early childhood origin, is a condition associated with increasing age. Thus, those whose viewing and appreciation of programmes might be enhanced by subtitles, in the main, do not have access to them. Among hard-of-hearing viewers who did have access to the teletext subtitle service, two thirds of those aged 51 years and over felt that subtitles assisted their understanding of television programmes. As one might expect, of those owning teletext, hard-of-hearing viewers reported greatest use of subtitles. Thirteen per cent of those with hearing difficulty and aged over 51 years said they used subtitles for all programmes watched and a further 26% of the over fifties with hearing difficulty reported regularly using subtitles for selected programmes. These data advocate that there are many hard of-hearing viewers whose viewing could be enhanced by subtitling. Effort should be made to promote public and professional awareness of the availability of teletext subtitling and additional resources will be needed to combat the technophobia which may hinder older people from initially purchasing a teletext television set, or making effective use of one they already own. PMID- 8241966 TI - Hearing loss risk from exposure to shooting impulses in workers exposed to occupational noise. AB - The effect of shooting impulses on hearing was analysed in 150 professional forest workers exposed to noise from chain saws. The exposure to shooting impulses (Lesi) was designed to take into account the peak levels of shooting impulses, their number and use of hearing protectors. Hearing loss was dependent on Lesi even after allowing for the age of subjects and their exposure to chain saw noise. Hearing threshold levels were compared between pairwise matched groups with high and low Lesi. Proper matching was achieved for age, chain saw noise, salicylate consumption, blood pressure, cholesterol, smoking and vibration induced white finger symptoms. The workers with high Lesi had 9 dB greater hearing loss at 4 kHz and 10 dB greater hearing loss at 8 kHz than those with low Lesi, the difference being significant at P < 0.05 level. In evaluation of noise induced hearing loss the exposure to shooting impulses from different calibre weapons should be determined since they increase the extent of hearing loss. The present study describes the Lesi method which more accurately evaluates the harmful effects of shooting noise impulses on hearing. PMID- 8241967 TI - Cochlear damage in guinea pigs following contralateral sound stimulation with and without gentamicin. AB - The effect of a minimally damaging sound exposure and a sub-ototoxic dose of gentamicin on cochlear hair cells contralateral to the sound exposure was evaluated. The cochleae of pigmented guinea pigs exposed to an 8 kHz pure tone at 116 dB SPL for 1 h and/or 50 mg/kg/day of gentamicin for 10 consecutive days and repeated after an interval of 3 weeks, were used for this purpose. Hair cell loss was found to have occurred in the contralateral cochleae following the sound exposure alone. The occurrence of potentiation, synergism and differential synergism between the agents in the contralateral ears was also seen. Possible explanations for these phenomena are proposed. PMID- 8241968 TI - Microslicing of the resin-embedded cochlea in comparison with the surface preparation technique for analysis of hair cell number and morphology. AB - The surface preparation technique for hair cell counting is frequently used for the analysis of cochlear pathology in relation to hearing impairment. To overcome problems encountered with standard surface preparation methods, a new combination of resin embedding and microslicing has been applied to the mammalian cochlea, to permit hair cell counting to be followed by electron microscopic analysis. Partial removal of the cochlear shell is followed by resin infiltration under vacuum prior to polymerization. The cochlea is then subdivided into hemicoils using a mounted annular diamond blade. Embedding before microslicing ensures that both apical and basal regions of the cochlea are preserved equally well with an evenly distributed and therefore predictable 10% hair cell loss. By comparison, the standard surface preparation method often produces distortion of remaining organ of Corti and unpredictable losses of hair cells, for example, up to 17%. The damage tends to be greatest towards the base of the cochlea, an area which shows particular susceptibility to ototoxic agents such as the aminoglycoside antibiotics. Thus, for assessment of pathology caused by ototoxic agents, this method has considerable advantages over the surface preparation technique. PMID- 8241969 TI - Auditory brainstem responses recorded at fast stimulation rates using maximum length sequences. AB - There are now commercially available systems which can provide maximum length sequence averaging techniques. This study examines the effects on auditory brainstem response amplitude and latency of using such a system and stimulating at rates from 9 to 1000 clicks/s. As expected there is considerable adaptation of the response such that at 1000 clicks/s wave V amplitude is only approximately 8% of its low stimulation rate value. As the stimulation rate is increased so the test time taken to obtain a response of the same signal-to-noise ratio as a conventional recording decreases. This process continues up to a maximum click rate of about 200 clicks/s. Thereafter, the response adapts so rapidly that further increases of rate lead to a worsening of performance. Thus, the optimum stimulation rate appears to be of the order of 200 clicks/s and this can give a speed improvement by a factor of approximately 2.8 over conventional recordings taken at 9 or 10 clicks/s. PMID- 8241970 TI - Correcting for factors affecting ABR wave V latency. AB - The latency of auditory brainstem response (ABR) wave V has been shown to be affected by several subject and technical factors. Previous studies have often concluded with a recommendation to account or correct for one or more of these factors when latency is used as a means of detecting retrocochlear dysfunction. This study investigated the effect of subject age, sex, hearing loss and stimulus intensity on ABR wave V latency in a group of 189 clinical subjects in whom retrocochlear disease was effectively excluded. Regression analysis showed that all four factors exerted a significant influence on latency and that hearing loss and intensity were most effectively represented when combined to form a sensation level variable, together with a measure of audiogram slope. PMID- 8241971 TI - Comparison of two methods of data presentation for the electrical brainstem response in cochlear implant patients. AB - Cochlear implants have become an acceptable method for management of the deaf patient, adult and child alike. Since the advent of implants there has been much research into objective methods for setting electrode values. The most common method is the electrical auditory brainstem response (EABR). The result of EABR threshold measurement is frequently reported as an arbitrary stimulus level set by the manufacturer. However, there has been recent discussion of whether an arbitrary stimulus level is acceptable or if the results should be reported in absolute units, either charge or current, particularly when combined data are used. This report compares two methods of data presentation for the EABR, arbitrary stimulus level and absolute units measured in nanoCoulombs (nC). Data collected from a previous study were used in this comparison. All patients were fitted with the Nucleus multi-channel device and Mini Speech Processor. The correlation between stimulus level and log charge (nC) was 0.990 with a slope of 0.987 which suggests the method of data reporting does not alter EABR results materially when using the Nucleus device. PMID- 8241972 TI - Effects of postural change on the middle and inner ear. PMID- 8241973 TI - Systemic lupus erythematosus-related autoantibody production in mice is determined by bone marrow-derived cells. AB - Experimental systemic lupus erythematosus (SLE) can be induced in mice by immunization with either a human monoclonal anti-DNA antibody bearing the 16/6 idiotype (16/6 Id) or with a mouse monoclonal anti-idiotypic antibody specific for the 16/6 Id. Susceptibility to the induction of experimental SLE is genetically determined but is not linked to the MHC. In the present study we tested the susceptibility of BM chimeras of different donor-host combinations to the induction of SLE and found that high levels of anti-16/6 Id and anti-ssDNA antibodies were induced in BALB/c-->C57BL/6, BALB/c-->BALB/c and normal BALB/c mice as opposed to C57BL/6-->BALB/c chimeras and normal C57BL/6 mice. The low levels of the anti-16/6 Id and anti-ssDNA antibodies produced by C57BL/6-->BALB/c chimeras immunized with the 16/6 fully allogeneic BMT as such chimeras were shown to produce high levels of antibodies to a T cell-dependent antigen (the synthetic polypeptide (Phe,G)-A--L). These results demonstrate that the production of SLE related autoantibodies is controlled by donor-type BM derived cells and not by host-type cells in the thymic stroma. PMID- 8241974 TI - Low-dose heparin inhibits acute graft versus host disease in mice. AB - We have studied the effect of low-dose heparin on GVHD, marrow engraftment and graft-versus-leukemia (GVL) effects in an experimental murine model. Recipient (C57BL/6 x BALB/c) F1 mice were transplanted with C57BL/6 marrow and/or spleen cells and treated with daily sc injection of 5 micrograms heparin for 30 days. We have shown that heparin in low doses attenuates the severity of acute GVHD and reduces the mortality rate from 69 to 37.5% without abrogating the GVL effect induced by the allograft and without impairing marrow engraftment. PMID- 8241975 TI - Merocyanine 540-sensitized photoinactivation of high-grade non-Hodgkin's lymphoma cells: potential application in autologous BMT. AB - This paper reports on the preclinical evaluation of merocyanine 540 (MC540) as an agent for the inactivation of tumour cells in BM grafts from non-Hodgkin's lymphoma (NHL) patients. The three cell lines used for this study, OCI-LY13.1, OCI-LY13.2 and OCI-LY9, originate from two patients with high-grade NHL. The OCI LY13.1 and OCI-LY13.2 lines are derived from the same patient. The OCI-LY13.1 line was established at the time of diagnosis while the OCI-LY13.2 line was established after the tumour had become refractory to therapy. When used under conditions that are known to preserve about 50% of normal human pluripotent hematopoietic progenitor cells (CFU-GEMM), MC540-sensitized photoirradiation reduced in vitro clonogenic OCI-LY9 cells by 4 orders of magnitude and OCI-LY13.1 and OCI-LY13.2 cells by > or = 5. Survival curves for OCI-LY13.1 and OCI-LY13.2 cells were similar and followed first order kinetics, while those for OCI-LY9 cells were distinctly biphasic. Suspension cultures established with photoinactivated lymphoma cells confirmed that MC540-sensitized photoirradiation was cytotoxic and capable of eliminating > or = 4 log of tumour cells. These results encourage the further exploration of MC540-sensitized photoirradiation as a means to purge autologous marrow grafts from NHL patients. PMID- 8241976 TI - Prophylactic use of ganciclovir for allogeneic bone marrow transplant recipients. AB - Ganciclovir which has proved effective in the treatment of cytomegalovirus (CMV) infection was given prophylactically to 40 bone marrow transplant (BMT) patients pre and post-transplant in seropositive patients and post-transplant in seronegative patients with a seropositive donor. All patients were transfused with screened blood products and 33 received CMV hyperimmune globulin. They were compared with an historical control group consisting of 39 patients who had received significantly more unscreened blood products (p = 0.01) and less HLA mismatched marrow transplants (p = 0.05). Toxicity of ganciclovir was hematological-neutropenia was responsible for cessation of the drug in seven patients and transfusion requirements were significantly higher in the ganciclovir group. Non-hematological toxicity did not occur in any patient. Only one patient (2.5%) experienced symptomatic CMV infection and no patient developed CMV pneumonitis. In contrast, in the control group, 23 (59%) patients had clinical symptoms of CMV infection (p < 0.0001) and 4 (10%) experienced CMV pneumonitis (p < 0.01). Ganciclovir significantly reduced the incidence of positive CMV antigenemia (7.5% in the treated group vs 72% in the control group; p < 0.01). However, ganciclovir delivery did not result in an improved overall survival due to a higher rate of regimen-related deaths and chronic GVHD mostly in patients transplanted from an HLA-mismatched donor. The prophylactic administration of ganciclovir abrogates CMV pneumonitis and considerably reduces the incidence of CMV infection in BM recipients at high risk of developing this disease after transplantation. PMID- 8241977 TI - Invasive fungal sinusitis in patients undergoing bone marrow transplantation. AB - Invasive fungal sinusitis is becoming increasingly common in patients undergoing BMT. This study was undertaken to evaluate the incidence, presenting symptoms, diagnosis procedures, treatment and outcome of invasive fungal sinusitis. The study population comprised 423 consecutive BMT patients at Hadassah University Hospital from January 1986 to August 1992. Eleven patients (2.6%) developed invasive fungal sinusitis, 8 had underlying hematologic malignancies and 3 severe aplastic anemia (SAA). Median interval between BMT and fungal sinusitis was 22.5 days (range 2-106 days). Eight of 11 patients had protracted neutropenia (median 8 days with median neutrophil count at the time of fungal sinusitis diagnosis of 0.25 x 10(9)/l). Four patients developed GVHD before fungal sinusitis was diagnosed. Presenting symptoms were fever (100%), orbital swelling (63%), facial pain (54%) and nasal congestion (36%). In 8 patients Aspergillus species were isolated (A. flavus in 7, A. quadrilineatus in 1); in 1 patient Candida albicans was isolated and in the other 2 fungal elements were detected histologically (Fusarium and Mucor, respectively). Six of the patients underwent surgical debridement at diagnosis. Three received granulocyte transfusions. All patients received systemic amphotericin B (7 conventional and 4 amphotericin B colloidal dispersion (ABCD)). Only 2 of the 11 patients responded completely to therapy with a follow-up of 15 months. It appears that invasive fungal sinusitis is a potentially fatal complication in immunocompromised patients post-BMT. Current treatment approaches are largely ineffective and new methods of management of this serious problem are needed. PMID- 8241978 TI - Graft rejection by a population of primed CDw52- host T cells after in vivo/ex vivo T-depleted bone marrow transplantation. AB - We investigated a case of graft rejection after in vivo/ex vivo T-depleted BMT in a patient who had received a HVG-matched, GVH one locus-mismatched, MLC-negative graft from his cousin. In vivo/ex vivo T cell depletion was performed with Campath 1G (CP1G) and Campath 1M (CP1M), respectively. We identified a failure of CP1G to eradicate a CDw52- (Campath-negative) host T cell population as the main cause of treatment failure. The analysis also suggests that significant host versus-donor reactivity prior to transplant, as detected by limiting dilution analysis, also contributed to graft rejection. The rejecting T cells were bifunctional in that they showed cytotoxic activity and were capable of inhibiting haemopoietic progenitor growth by producing inhibitory lymphokines. PMID- 8241979 TI - Pentostatin increases the acute toxicity of high dose cyclophosphamide. AB - One dose of pentostatin was added to a standard cyclophosphamide (CY) based transplant regimen in two patients in an attempt to decrease the rate of non engraftment in haploidentical allogeneic BMT. Despite a normal cardiac history and evaluation prior to transplant, both patients suffered fatal cardiac toxicity within 48 h of receiving the chemotherapy. This phenomenon was further investigated in an animal model. Laboratory rats were treated with progressive doses of CY in a range that produces acute cardiac toxicity. Successive groups of rats were treated with either pentostatin or fludarabine and CY at 400 mg/kg. Neither pentostatin nor fludarabine alone produced early mortality. However, a marked increase in early mortality was noted in those animals treated with pentostatin and high-dose CY. The addition of fludarabine did not increase the early toxicity of CY. Autopsy revealed no gross or microscopic abnormalities in the animals. The implications of adding agents that interfere with adenosine metabolism to CY based transplant regimens is discussed. PMID- 8241980 TI - Evaluation of chimerism after bone marrow transplantation with single locus minisatellite DNA probes. AB - The origin of cells in almost all allogeneic donor-recipient pairs can be determined through the use of highly polymorphic minisatellite DNA probes. Single locus probes were cloned from hypervariable fragments in a human DNA fingerprint detected with a multi-locus probe. While each probe is highly polymorphic and locus specific, they all contain repetitive sequences. The properties of single locus probes have improved the sensitivity of detecting mixed chimerism in comparison with multi-locus probes. The use of single-locus probes permitted detection of mixed chimerism (MC) at levels as low as 0.625%, approaching that obtained by PCR methods. In the present study, five patients who received allogeneic BMT for hematologic malignancies were analyzed. Two patients exhibited MC after BMT. One developed acute GVHD and chronic GVHD and remained in CR while the second patient who had no signs of GVHD suffered a relapse. PMID- 8241981 TI - Acute graft-versus-host disease and the risks for idiopathic pneumonia after marrow transplantation for severe aplastic anemia. AB - The risks for the development of idiopathic pneumonia after allogeneic BMT were assessed in a case-series review at a single marrow transplantation center. All allogeneic marrow recipients (n = 299) (age range 1-60 years) with severe aplastic anemia (SAA) transplanted from family member donors after conditioning with CY were evaluated. Post-grafting immunosuppression consisted of MTX alone in 205 patients (69%), CY alone in 16 (5%) and a combination of the two in 78 (26%). The incidence estimate for any pneumonia within the first 200 days after transplant was 18% (95% confidence interval = 14-24%). Of 48 cases of pneumonia, CMV infection was documented in 44%, 21% were idiopathic and the remainder were either due to other infections or were not evaluated. The effect of acute GVHD on the incidence of pneumonia was examined using multivariate Cox proportional hazards models which included covariates for potential confounding factors. Consistent with previous reports, acute GVHD was associated with an increased incidence of any pneumonia (relative risk (RR) = 3.5, 95% Cl = 1.9-6.9; p < 0.001). Specifically, acute GVHD also was associated with the largest risk of idiopathic pneumonia (RR = 5.0, 95% Cl = 1.1-22; p = 0.04). In conclusion, recognition of acute GVHD as a risk factor for idiopathic pneumonia suggests that mechanisms in addition to chemoradiation damage are responsible for non infectious lung injury after BMT. PMID- 8241982 TI - Incidence of cataracts after single fraction total body irradiation: the role of steroids and graft versus host disease. AB - Eighty-eight patients who received single fraction total body irradiation (sfTBI) as part of their conditioning for allogeneic BMT have been evaluated for the risk of cataract formation. Thirty-eight (43%) have developed cataracts; 11 required surgery. With 9.5-13.6 years follow-up (median 10.7 years), all 12 recipients of unmanipulated marrow allografts have developed cataracts; the actuarial risk of needing surgery was 32 (+/- 18%, 95% confidence intervals (CI)). Ten of these 12 required high-dose steroids (prednisolone > 1 mg/kg/day) for the treatment of GVHD. Seventy-six patients received T cell-depleted allografts; 14 of 76 required post-transplant immunosuppression with high-dose steroids. With 1-9.4 years follow-up (median 5 years), the actuarial risk of cataract formation in T cell depleted allograft recipients is 72% (+/- 52% CI), the actuarial risk for needing surgery is 20% (+/- 9% CI). Recipients of sfTBI and non-T cell-depleted allografts had a significantly greater risk of developing cataracts (p = 0.003, long rank test) and of needing surgery (p < 0.05, log rank test) than patients receiving T cell-depleted BM. Cataracts occurred more frequently in patients requiring post-transplant immunosuppression with steroids (relative risk 2.12, p < 0.01 log rank test). PMID- 8241983 TI - Invasive pulmonary aspergillosis prior to BMT in acute leukemia patients does not predict a poor outcome. AB - Eight patients with acute leukemia (AL) and invasive pulmonary aspergillosis (IPA) developing during previous antileukemic therapy underwent BMT (autologous in 6 cases and allogeneic 2). IPA was treated prior to BMT with full doses of amphotericin B, associated with surgical resection in three cases. One patient was treated with amphotericin B and itraconazole. Prior to BMT, seven patients had minimal residual pulmonary lesions. All patients received amphotericin B (0.5 mg/kg/day) during the aplastic period prior to engraftment. One patient died of Gram-negative septic shock before engraftment. Seven patients achieved complete hematological engraftment without any evidence of IPA reactivation. Amphotericin B was well tolerated with only minimal transient renal dysfunction in three patients. Later pulmonary complications related to IPA were observed in only one patient who developed a self-limited episode of hemoptysis. One patient died of CMV pneumonitis and two of leukemia relapse. Four patients survive disease-free and without complications related to IPA. We conclude that the reactivation of correctly treated IPA can be successfully prevented in BMT patients by use of prophylactic amphotericin B. With this approach, prior IPA is not a contraindication to BMT. PMID- 8241984 TI - Monoclonal antibody-purged autologous bone marrow transplantation in adults with acute lymphoblastic leukemia at high risk of relapse. AB - The prognosis for adults with B lineage ALL who have relapsed after an initial remission is poor. High-dose chemoradiotherapy followed by autologous BMT can induce prolonged clinical remissions in some children with recurrent ALL. In this study, we evaluated the efficacy of autologous BMT in adults. Autologous marrow was treated in vitro with J5 and J2 monoclonal antibodies (CD10/CD9) plus rabbit complement to purge residual ALL cells. Twenty-two adults with B lineage ALL were treated with high-dose chemoradiotherapy followed by infusion of J2/J5 purged autologous BM. The median age was 28 years (range 18-54 years). Twenty-one of 22 patients had experienced at least one relapse prior to BMT. All patients achieved complete hematologic engraftment. Disease-free survival (DFS) in this cohort of patients was 20%, with all survivors alive and free of disease between 2.5 and 7.5 years post-BMT. Age at the time of BMT was an important prognostic factor, with patients < 28 years old faring much better than older individuals (DFS, 45% vs 0%, p = 0.01). Our experience suggests that high-dose chemoradiotherapy followed by infusion of J2/J5 purged autologous marrow is as efficacious in young adults as it is in children and is a reasonable alternative for patients who lack HLA-matched donors. Results in older adults are poor, however, and demonstrate the need for more effective transplant strategies in these individuals. PMID- 8241985 TI - Hypercoagulability in patients undergoing autologous or allogeneic BMT for hematological malignancies. AB - Severe thrombotic alterations, such as veno-occlusive disease of the liver, may occur in the early phase following high-dose chemoradiotherapy and BMT. In this study, performed in patients with hematological malignancies subjected to allogeneic (10 cases) and autologous (20 cases) BMT, we have monitored laboratory hemostatic parameters to better understand the pathogenetic mechanism of thrombosis and particularly of veno-occlusive disease. Prothrombin time, activated partial thromboplastin time, plasma fibrinogen, markers of hypercoagulability (thrombin-antithrombin complex and prothrombin fragment F1+2); natural anticoagulants (protein C, protein S and antithrombin) together with fibrinolytic parameters (plasminogen, alpha 2-antiplasmin, tissue-plasminogen activator, plasminogen activator inhibitor and D-dimer) were assessed before transplant, on day 0 and weekly for 1 month thereafter. A hypercoagulability state, not related to an impairment of the anticoagulant and fibrinolytic systems, was documented before and after autologous and allogeneic transplant. Two patients developed veno-occlusive disease: they did not show any difference from the other patients before transplant while they presented a decrease of the natural anticoagulants along with altered fibrinolytic parameters only at the clinical onset of veno-occlusive disease. In conclusion, in this study a state of marked hypercoagulability was documented in BMT patients and the hemostatic laboratory parameters evaluated were not able to predict the occurrence of the thrombotic complications. PMID- 8241986 TI - Elimination of clonogenic Philadelphia-positive cells using BCR-ABL antisense oligodeoxynucleotides. AB - Synthetic BCR-ABL antisense oligodeoxynucleotides are known to suppress in vitro clonogenic growth of primary cells from patients with CML. To evaluate the use of BCR-ABL antisense oligodeoxynucleotides as in vitro purging agents before autografting, we studied their effect on the clonogenic BV-173 cell line. At a concentration of 80 micrograms/ml, antisense oligodeoxynucleotides suppressed 85%, 95.5% and 95% of leukaemic cell growth after 24, 48 and 72 h of incubation, respectively. A correlation was observed between the concentration of oligomers and cell inhibition; however, concentrations higher than 80 micrograms/ml did not produce significant increase of BV-173 cell elimination. When the antisense treatment was performed on a wide range of target cell concentrations (from 1 x 10(5) to 5 x 10(6)/ml), the efficacy of purging was similar in all the groups. Addition of antisense oligodeoxynucleotides every 24 h produced, after 3 days, a cell growth inhibition superior to any single treatment; in particular, three treatments for 24 h at a concentration of 80 micrograms/ml were more effective than a single treatment at the same concentration or a single treatment at a concentration of 160 micrograms/ml. Incubation of normal human BM cells with the same doses which were very effective at inhibiting BV-173 cell proliferation, did not affect the growth of normal CFU-GM and of more immature progenitors. PMID- 8241987 TI - Efficacy of immune globulin in preventing complications of bone marrow transplantation: a meta-analysis. AB - To assess the efficacy of immune globulin in preventing CMV infection, interstitial pneumonia, GVHD and death after BMT, we reviewed and synthesized data from 12 published studies (with 1282 patients) in which immune globulin was used prophylactically in BMT patients, controls were included and clinical outcomes were assessed. Data synthesis indicates that immune globulin significantly reduces fatal CMV infection (odds ratio (OR) 0.47, 95% confidence interval (CI) 0.23-0.99), CMV pneumonia (OR 0.61, CI 0.42-0.89), non-CMV interstitial pneumonia (OR 0.57, CI 0.35-0.95) and total mortality (OR 0.74, CI 0.55-0.99). The reduction in acute GVHD was not quite significant (OR CI 0.45 1.02). Complications decrease with both hyperimmune and conventional immune globulin. For CMV-negative transplant recipients, immune globulin decreases symptomatic CMV infection (OR 0.55, CI 0.31-0.94) and interstitial pneumonia (OR 0.34, CI 0.15-0.77). For CMV-positive recipients, immune globulin prevents interstitial pneumonia (OR 0.45, CI 0.26-0.80) but not symptomatic CMV infection (CI 0.41-2.80). We conclude that immune globulin is efficacious in preventing major complications of BMT in both CMV-negative and CMV-positive recipients. PMID- 8241988 TI - Intravenous immunoglobulin and CMV-seronegative blood products for prevention of CMV infection and disease in bone marrow transplant recipients. AB - The efficacy of i.v. immunoglobulin plus CMV-seronegative blood products or CMV seronegative blood products alone for prevention of CMV infection, symptomatic CMV disease, other infections and GVHD after BMT was evaluated in a randomized, controlled trial. Fifty-one CMV-seronegative allogeneic BMTs with a CMV seronegative or CMV-seropositive marrow donor were randomly assigned to receive either i.v. immunoglobulin (1.0 g/kg once weekly for 120 days after transplant) plus CMV-seronegative blood products or CMV-seronegative blood products alone. CMV infection occurred in 2 of 25 patients (7%) receiving i.v. immunoglobulin plus CMV-seronegative blood and in 2 of 23 patients (9%) receiving CMV seronegative blood alone. All CMV infections were asymptomatic and characterized by viral excretion with or without CMV seroconversion. There were no cases of CMV related interstitial pneumonia. Grade > or = II GVHD was less frequent in patients given i.v. immunoglobulin (5 of 25 patients (20%) vs. 11 of 23 patients (48%), p = 0.04). The number of bacterial and fungal infections was similar in both groups. Fewer non-CMV viral infections (9 of 27 patients (33%) vs. 15 of 24 patients (63%), p = 0.03) and fewer deaths associated with infection (1 of 27 patients (4%) vs. 5 of 24 patients (21%), p = 0.07) occurred in recipients of immunoglobulin. Neither survival nor risk of leukemia relapse was changed by the immunoglobulin. The high doses of i.v. immunoglobulin were well tolerated. These results suggest that CMV-seronegative blood products alone prevent most CMV infections and CMV disease in CMV-seronegative allogeneic BMT recipients, even when the marrow donor is CMV-seropositive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8241989 TI - Effect of GVHD on the recovery of NK cell activity and LAK precursors following BMT. AB - The mechanism by which GVHD augments the graft-versus-leukemia (GVL) effect of marrow transplants has not been ascertained. One possibility involves the secondary activation of natural killer (NK) cells by cytokines released during the GVHD process. To evaluate this possibility we have compared NK activity and lymphokine-activated killer cell precursor (LAKp) frequencies in serially sampled PBMC from recipients of unmanipulated autologous or allogeneic marrow with and without active GVHD. NK activity recovered rapidly after BMT and was elevated during episodes of acute GVHD. However, NK activity did not differ between recipients of autologous or allogeneic marrow without GVHD nor was NK activity increased in association with chronic GVHD. Endogenously-activated NK cells were detected only in recipients of allogeneic marrow but this did not correlate with GVHD status. In contrast to NK activity, LAKp frequencies fell below the control range during the first 8 weeks after BMT. By 9-14 weeks the median LAKp frequency was normal and did not differ between the three groups then or later after transplant. We conclude that acute GVHD may serve to increase the lytic activity of NK cells but does not result in increased LAKp. LAKp frequencies are below normal during the first two months after BMT, a finding not previously recognized from bulk culture LAK studies. The role of LAK effectors in GVL may involve more the degree of cellular activation rather than the number of cells activated. PMID- 8241990 TI - Diffuse alveolar hemorrhage following autologous bone marrow infusion. AB - Autologous BMT performed in a 57-year-old woman with relapsed large cell lymphoma was complicated by two consecutive episodes of diffuse alveolar hemorrhage (DAH). The second episode occurred immediately after infusion of autologous BM. DAH is an increasingly recognized complication of autologous BMT and carries a high mortality. It is characterized by dyspnea, cough, bilateral pulmonary infiltrates and progressively bloodier aliquots of bronchoalveolar lavage fluid. The pathogenesis is probably multifactorial involving an initial insult to lung endothelium with inflammatory cells serving as the mediators of subsequent injury. The rapid development of DAH following marrow infusion strongly implicates DMSO as a potential cause in our patient. PMID- 8241991 TI - Berlin Stem Cell Workshop. April 30 and May 1, 1993. Abstracts. PMID- 8241992 TI - Radioisotopic labels for blood cell survival studies: a review. AB - The different methods of labeling blood cells, in particular erythrocytes and platelets, for survival studies are reviewed. Besides the technical information about these procedures, attention is also paid to the indications of blood cell survival studies, the effective dose equivalent involved and the interpretation of the resulting data. In the final section the possibilities created by dual labeling procedures are discussed. PMID- 8241993 TI - Renal elimination of some 99mTc-labelled cysteamine derivatives. AB - It is generally believed that -CO-NH-(CH2)n-COOH moiety promotes tubular excretion of organic anions. Recently it was reported that newly developed renal agents, containing the oxotechnetium(V) glycine group, may mimic the carbonyl amide sequence of [131]I-o-iodohippuric acid. In this study the renal excretion of certain carboxy-cysteamine derivatives is investigated in mice, in the presence of renal tubular transport inhibitor. A similar pattern of renal depression was observed for complexes containing the oxotechnetium glycine sequence, suggesting that this group may satisfy structural parameters for tubular secretion of anionic technetium complexes. PMID- 8241994 TI - The labeling of proteins and LDL with 99mTc: a new direct method employing KBH4 and stannous chloride. AB - A new direct labeling technique for proteins with technetium-99m (99mTc) has been developed and makes use of borohydride and stannous chloride. The method is simple and reproducible and gives a high labeling efficiency and high retention of biological activity for proteins, including polyclonal immunoglobulin (Ig), antifibrin monoclonal antibody, tissue type plasminogen activator, fibrinogen and low density lipoprotein (LDL). This method can be used in kit-format and takes about 20 min preparation time at room temperature. Both in vitro and in vivo studies indicate good stability of the label. In vivo, mice and rabbit images show significant accumulation of 99mTc-Ig at an inflammation area, 99mTc antifibrin at a thrombus site and 99mTc-LDL in atherosclerotic lesions. The method is attractive for routine research and clinical purposes. PMID- 8241995 TI - Radioiodination of a monoclonal antibody using N-succinimidyl 5-iodo-3 pyridinecarboxylate. AB - The potential utility of N-succinimidyl 5-iodo-3-pyridinecarboxylate (SIPC) for the radioiodination of monoclonal antibodies was investigated. Paired-label studies were performed using the anti-tenascin antibody 81C6 in athymic mice bearing subcutaneous D-54 MG human glioma xenografts. Radiolabeling was also done using N-succinimidyl 3-iodobenzoate (SIB). Radioiodination of SIPC and SIB both proceeded in 60-80% yield, but protein coupling efficiencies with SIB were higher (76 +/- 16 vs 60 +/- 7%). Immunoreactivity and affinity of both preparations were similar. Using SIPC, thyroid uptake was quite low, decreasing from 0.3% at day 1 to 0.05% at day 8. Tumor uptake reached 46 +/- 11% injected dose/g at day 1 but declined gradually thereafter. This apparent decline reflected the rapid growth of these xenografts since tumor accumulation expressed as percentage of injected dose remained nearly constant up to day 9. These results suggest that SIPC, like SIB, offers significant advantages for labeling antibodies when compared with conventional protein iodination methods. PMID- 8241996 TI - Synthesis and characteristics in tumor-bearing mice of N-[11C]methyl-1 deoxynojirimycin and N-[11C]methyl-1-deoxymannojirimycin. AB - N-[11C]Methyl-1-deoxynojirimycin ([11C]MDNM) and N-[11C]methyl-1 deoxymannojirimycin ([11C]MDMM) were prepared by 11C-methylation of 1 deoxynojirimycin (DNM) and 1-deoxymannojirimycin (DMM), which are specific inhibitors of glucosidase and mannosidase, respectively. In mice bearing Ehrich ascitic tumor, the highest uptake of the [11C]MDNM was observed in the kidney, followed by the liver and small intestine, while the tumor uptake was moderate. By MDNM loading, saturable uptake was observed in these tissues. In homogenates of the kidney and tumor tissues, a considerable amount of radioactivity was detected in a high-molecular weight fraction. These results demonstrate that the [11C]MDNM has a potential for imaging the glucosidase activity by positron emission tomography. On the other hand, [11C]MDMM showed lower uptake than [11C]MDNM in the kidney, liver and small intestine and no effect of carrier DMM, suggesting that the [11C]MDMM would not reflect mannosidase activity. PMID- 8241997 TI - Radioimmunodetection of rat and rabbit cartilage using a monoclonal antibody specific to link proteins. AB - Biodistribution analysis using [125I]Fab-6F3 specific to link proteins from human articular cartilage performed in rats by autoradiography showed a high concentration of radioactivity in all cartilaginous tissues. Preliminary immunoscintigraphic assays were performed in rabbits. Front and side view images of whole animals exhibited high uptake in cartilage tissue of the knee articulation, in the invertebral disk and the humeral head. This fixation was still detected 24 h post-injection, although high washout of radioactivity was observed. PMID- 8241998 TI - Potential 99mTc radiopharmaceuticals for renal imaging: tris(N-substituted-3 hydroxy-2-methyl-4-pyridinonato)technetium(IV) cations. AB - A series of monocationic complexes of N-substituted-3-hydroxy-2-methyl-4 pyridinones labeled with technetium(IV)-99m have been evaluated in vivo as potential radiopharmaceuticals. The pyridinones have different substituents at the ring nitrogen atom: ethyl, i-propyl, i-butyl, benzyl, phenyl, p methoxyphenyl, 3-butoxypropyl and cyclohexyl. Biodistribution studies of the 99mTc complexes have been carried out in rabbits and mice. High kidney uptake and retention of the radionuclide has been shown in rabbits and mice with the cationic complexes of 3-hydroxy-1-(p-methoxyphenyl)-2-methyl-4-pyridinone and 1 (cyclohexyl)-3-hydroxy-2-methyl-4-pyridinone. These 99mTcL3+ compounds appear to be morphologic renal agents. PMID- 8241999 TI - Synthesis and evaluation of 1-(1-[5-(2'-[18F]fluoroethyl)-2-thienyl] cyclohexyl)piperidine as a potential in vivo radioligand for the NMDA receptor channel complex. AB - 1-(1-[5-(2'-[18F]Fluoroethyl)-2-thienyl]cyclohexyl)piperidine (18FE-TCP) was prepared as a fluorine-substituted analogue of the potent NMDA receptor channel blocker, 1-[1-(2-thienyl)cyclohexyl]piperidine (TCP), by the mesylate displacement with [18F]fluoride ion with isolated radiochemical yields of 6-12%, and the synthesis time including a two step HPLC purification was 120 min. The regional distribution in rat brain after i.v. injection of 18FE-TCP was heterogeneous and similar to the known distribution of phencyclidine recognition sites, with hippocampus-cerebellum, striatum-cerebellum and cerebral cortex cerebellum concentration ratios of 2.08, 1.7 and 1.54, respectively, 15 min post injection. Furthermore, this localized regional cerebral distribution was blocked by co-injection with the unlabelled FE-TCP or pretreatment with cis-2 hydroxymethyl-r-1-(N-piperidyl)-1-(2-thienyl)cyclohexane, with the greatest reductions seen in the hippocampus followed by the striatum and cerebral cortex. However, relatively low receptor binding affinity and high non-specific binding due to its high lipophilicity suggest that 18FE-TCP may not be a suitable radioligand for in vivo PET investigations of the NMDA receptor-channel complex. PMID- 8242000 TI - First-pass studies of acute lung injury. AB - Mild hydrochloric acid was introduced to a caudal lung section in each of eight dogs to induce injury. Transits of 99mTc-labeled red blood cells (RBC) and [123I]iodoantipyrine (IAP) injected intravenously were recorded by a scintillation camera. Lungs and blood samples were analyzed post-mortem. Peak-to equilibrium ratios (P/E) of RBC time-activity curves were computed to be 3.83 +/- 0.54 for the control lung, 2.58 +/- 0.55 for the injured lung and 2.23 +/- 0.58 for the injured caudal section. For IAP, the respective results were 3.78 +/- 0.29, 2.02 +/- 0.18 and 1.77 +/- 0.17. The decrease of P/E in injured areas was attributed to reduced blood flow. Using mean transit times of the tracers, we computed extravascular lung water per unit blood volume to be 0.35 +/- 0.18 for the control lungs and an increased value of 0.68 +/- 0.24 for the injured lungs. These results displayed sensitivity to injury, but were gross underestimates relative to the corresponding values of 2.04 +/- 0.54 and 4.56 +/- 1.85 in post mortem analyses. PMID- 8242001 TI - 99mTc-citrate versus 67Ga-citrate for the scintigraphic visualization of inflammatory lesions. AB - Citric acid was labeled with 99mTc with an efficiency of > 99%. The biodistribution of 99mTc-citrate was studied in mice with turpentine-induced abscesses in comparison to 67Ga-citrate. The max. abscess/muscle concentration ratios were 4.61 +/- 1.92 (3 h) for 99mTc-citrate and 4.76 +/- 2.04 (4 h) for 67Ga-citrate. Arthritis was induced in 10 rabbits by intra-articular injection of ovalbumin Scintigrams obtained 4 days later and at 3 h post-injection of 99mTc citrate showed increased activity involving the synovium. The max. arthritic/contralateral knee ratio was 3.19 +/- 1.29 (3 h) and 6.47 +/- 3.71 (24 h) for 99mTc- and 67Ga-citrate, respectively. The blood clearance curve of 99mTc citrate in rabbits was biexponential with a fast (T1/2 = 36 min) and a slow (T1/2 = 18 h) component, compared to mono-exponential clearance of 67Ga-citrate (T1/2 = 23 h). In 10 patients with rheumatoid arthritis whole-body scintigrams and spot images of involved joints indicated localization of the tracer in inflamed tissues. The mean target-to-soft tissue ratios were 3.04 +/- 0.81 and 4.95 +/- 2.56 for 99mTc-citrate and 99mTc-MDP, respectively. Renal clearance of radioactivity was evident from the scintigrams. Our results demonstrated that 99mTc-citrate is effective as a radiopharmaceutical for the visualization of inflammatory lesions and may be preferred to 67Ga-citrate due to the ideal physical characteristics of the radionuclide, easy preparation, low cost, early accumulation and the preference for the renal route of excretion. PMID- 8242002 TI - Synkavit and its direct labelling with iodine-125, as a potential anti-cancer drug. AB - The compound known as "synkavit" is a diphosphate derivative of vitamin K3 (menadion), which is capable of being selectively accumulated in certain tumour cells, and covalently bonded to DNA producing considerable DNA damage. On the other hand, iodine-125 nuclide incorporated into the nucleus of living cells causes extreme radiotoxic effects. Consequently, synkavit can be used as a specific carrier of iodine-125 into the nucleus of tumour cells. Thus, its iodo derivatives have become interesting agents on the potential application of iodine 125 in cancer therapy. 6-Iodo-synkavit is a unique iodo-derivative described in the literature. In addition, its synthesis and radioiodination is still problematic, and consequently the results obtained using 6-iodo-synkavit labelled with iodine-125 remains in question. For this reason, the synthesis of 6-iodo synkavit was examined in this study. It is finally determined that a mixture of different iodo-isomers of synkavit has been produced rather than its specific 6 iodo-isomer, when the synthetic sequence was begun with the direct sulfonation of 2-methyl-naphthalene. On the other hand, it is also determined that synkavit can directly be radioiodinated using different iodination techniques, and iodogen especially can be successfully used as an oxidative agent. PMID- 8242003 TI - Physiological significance of myosin phosphorylation in skeletal muscle. AB - Each S-1 or head portion of the myosin molecule in skeletal muscle contains a subunit known as the regulatory or phosphorylatable light chain (P-LC). Phosphorylation of the P-LC is mediated by the second messenger Ca2+ and takes place when the muscle fibre is activated. In smooth muscle, phosphorylation of the P-LC is the principal mechanism that initiates contraction, but in skeletal muscle myosin P-LC phosphorylation is not required for contraction and a definitive role has not been established. It has been proposed that P-LC phosphorylation modulates the intrinsic nature of actin-myosin interactions, leading to force potentiation under suboptimal activation conditions. An example of this is posttetanic potentiation. This paper describes a P-LC phosphorylation induced mechanism for force enhancement during isometric contraction. In addition, it summarizes recent data revealing that P-LC phosphorylation is associated with enhanced work output of fast-twitch muscle during shortening and lengthening contractions. PMID- 8242004 TI - Determinants of independence in the elderly. AB - The purpose of this study was to compare strength, flexibility, and cardiorespiratory fitness of the elderly living independently (n = 64) with those in rest or nursing homes (n = 61), and to identify those variables most strongly associated with an independent lifestyle. Quantitative levels of independence (Incapacity Index) and of activity were determined by questionnaires. Flexibility was measured using the Leighton Flexometer. Strength was measured using a modified sphygmomanometer. Cardiorespiratory fitness was measured using a self paced walking test. Body size factors were also determined. Independent and dependent subjects were significantly different in age, which imposed a limitation on interpretation of these data. However, this factor was accounted for statistically in the analyses. Independent individuals compared to dependent ones showed significantly greater flexibility, activity levels, and choice of walking speed. Through multiple regression analysis, it was found that 40% of the variance in Incapacity Index was associated with participation in outdoor activities, greater shoulder flexibility, and a faster normal walking speed (> 1.0 m.s-1). PMID- 8242005 TI - Metabolic cost of ice and in-line skating in Division I collegiate ice hockey players. AB - This study compared the metabolic cost of ice skating and in-line skating in Division I collegiate hockey players. Heart rate and oxygen uptake were compared between the two conditions at three skating velocities: 12.5 km.h-1, 16.5 km.h-1, and 20 km.h-1. Twelve subjects were tested on two occasions: on ice (ice skating) and off ice (in-line skating). They skated for 3 min at each velocity, with heart rate recording and gas collection taking place during the final 30 s of each stage. A two-factor repeated measures ANOVA revealed that the chosen velocities affected the two conditions differently for heart rate (p < .01). No interaction between condition and velocity was found for absolute (p < .43) and relative (p < .15) oxygen uptake. In-line skating produced significantly greater heart rate values and absolute oxygen uptake values than ice skating at all three velocities (p < .05). In-line skating also generated significantly greater relative oxygen uptake values at 16.5 km.h-1 and 20 km.h-1. Results suggest that the metabolic cost of in-line skating is greater than that of ice skating for collegiate ice hockey players when skating at three velocities similar to those skated during game conditions. PMID- 8242006 TI - Anthropometric and computed tomographic assessment of the thigh in young and old men. AB - Anthropometric (AP) and computed tomographic (CT) methods of determining limb and muscle-plus-bone cross-sectional area (CSA) and volume (Vol) were compared in 13 young (M = 24.5 yrs) and 11 old (M = 71.0 yrs) men. CSA of total thigh, muscle plus-bone, quadriceps, and hamstring muscle compartments and muscle were measured from CT scans. Corresponding muscle Vol were estimated from anthropometric equations. Prediction equations for CT measures were developed from AP measures using multiple linear regression. AP and CT techniques produced different values for thigh component CSA and Vol, especially in the old men. AP overestimated muscle-plus-bone CSA and Vol (4-6%) and underestimated skin and subcutaneous tissue CSA and Vol (17-33%). Prediction equations for quadriceps CSA and Vol (R2 = 80-96%) were more accurate than equations for hamstrings (R2 = 42-65%). Specific thigh muscle CSA and Vol can be predicted from AP measures (SEE 5-15%). These findings may have clinical significance when normalizing strength per unit of muscle size. PMID- 8242007 TI - Torque/velocity properties of human knee muscles: peak and angle-specific estimates. AB - Angle-specific (AS) torque/velocity data have been used to avoid angle related variation in peak torque capacity. However, series elastic structures cause the contractile velocity of active force-producing tissue to differ from external joint velocity except at peak torque. Alternatively, angle related variation may be removed by normalizing peak torque to the isometric maximum at that angular position. The AS, peak (P), and normalized peak (NP) methods were compared in isovelocity knee flexion and extension at velocities between 50 and 250 degrees s 1 for 8 male subjects. The P and NP methods gave more similar torque/velocity relations than the AS method. Further, very little variation in peak torque was attributed to differences in joint angle. Both the P and AS methods illustrate that relative quadriceps/hamstrings torque capability (flexor/extensor ratio) increases slightly with velocity. It is proposed that antagonist muscle torque capabilities should be compared at different angular positions to assess muscular imbalance. PMID- 8242008 TI - Physiological effects of weight cycling in female lightweight rowers. AB - Repeated cycles of weight loss and regain are referred to as weight cycling. It is a practice of many athletes who must achieve a low body weight. The purpose of this study was to determine whether a history of weight cycling results in sustained metabolic changes. Healthy female rowers with a history of dieting and weight fluctuation (n = 7) were compared to a control group of rowers who had never dieted (n = 7). Anthropometric and metabolic measurements were done at pre , peak, and off-season during a 1-year period. At peak season the weight cyclers restricted their food intake and lost 4.2 +/- 1.8 kg, and subsequently regained 4.0 +/- 2.1 kg in the off-season. This was different from the controls (p = .003), who maintained a stable body weight at all times. No other group differences were observed. Resting metabolic rate (RMR) and triiodothyronine (T3) changed with time (p = .001, p = .000, respectively) in both groups, which appeared to reflect changes in fat free mass (FFM), not body weight. Long-term metabolic changes were not observed in these athletes. PMID- 8242009 TI - Evaluation of the VMM turbine for spirometry in the applied physiology laboratory. AB - The volume measurement module turbine (VMM) was evaluated in 51 subjects for spirometry in applied physiology against the Stead-Wells spirometer (SW) and Wright peak flow meter (WM). The volume and flow ranges (VMM) were, FEV1 1.32 to 3.94 L (mean 2.62, confidence interval [CI] 2.46 to 2.78); forced vital capacity (FVC) 1.97 to 5.06 L (mean 3.50, CI 3.29 to 3.71); and peak expiratory flow rate (PEFR) 290 to 624 L.min-1 (mean 434, CI 407 to 461). The mean difference for FEV1 was 0.09 L (CI 0.05 to 0.14), FVC 0.04 L (CI -0.02 to 0.10), and PEFR 18.0 L min 1 (CI 8.7 to 27.3) less than SW or WM. Bias with FEV1 and FVC was not significant, though PEFR demonstrated a significant proportional error. The repeatability coefficients for FEV1 and FVC were 0.18 and 0.20, comparable to the SW; but for PEFR they were greater, 58.4 versus 33.8 L.min-1 by WM. The VMM turbine is accurate and reliable for the measurement of FEV1 and FVC over the ranges studied; however, care should be taken when interpreting PEFR. PMID- 8242010 TI - Influenza B virus strain identification and the rise of influenza A/Beijing/32/92 like strains during the 1992-1993 influenza season in Canada. PMID- 8242011 TI - Influenza in Canada, 1992-1993 season. PMID- 8242012 TI - Bone regeneration guided by resorbable collagen membranes in rabbits: a pilot study. AB - Artificial cross-shaped intrabony defects were created in the mandibles of 12 rabbits and the cavities covered with Type I highly cross-linked resorbable collagen membranes for 30 days. Similar cavities were prepared in three control animals and left uncovered for the same time period. Morphological and analytical data were obtained by means of light microscopy, scanning electron microscopy, and x-ray energy-dispersive spectrometry. After the experimental period, the membrane covered cavities were completely filled with regenerated bone. In the control specimens, the artificial cavities were occupied by fibrous connective tissue. PMID- 8242013 TI - Finite element analysis of two-abutment Hader bar designs. AB - Parametric three-dimensional finite element studies were conducted on the Hader bar to determine mechanical properties with respect to bar length, stiffener height, and material properties. Three bar lengths (6, 12, and 18 mm) were analyzed with a 1-mm stiffener height. For the longest bar length (18 mm), two additional stiffener heights were evaluated (2 and 3 mm). In addition, three material types were studied for each of the design cases which represented a selection of alloys commonly used clinically. The model consisted of a representative Hader bar which was constructed of a bar of 1.8 mm diameter. The ends of the bar were fixed to a 2.5-mm diameter coping which was attached to a 1.8-mm diameter root form-type implant, rigidly fixed in a representative block of bone. The bone was modeled as fixed at its distal end to eliminate rigid body motion. A 200-Newton occlusal force was modeled as being transmitted through one, two, or three attachment clips, 5 mm in length, for the three respective bar lengths. The results of these analyses did not predict yielding (failure) for any of the cases studied. Span length and stiffener height were found to play a stronger role in the adequacy of the overall design as compared with changing material properties in the range of alloy stiffness tested. For the span length studies, factors of safety, with respect to static yield strength, ranged from 2.93 to 10.3 and fatigue factors of safety ranged from 1.41 to 3.36.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242014 TI - Polishing sequence for titanium using dental armamentarium: a pilot study. AB - With the increasing use of titanium in dentistry, the methods for maintenance, particularly the polishing sequence, have become especially important. Thirty test samples of commercially pure grade 2 titanium were sandblasted with 110 microns aluminum oxide at 60 psi to establish a baseline surface roughness. Two samples were exposed to each of 15 different polishing sequences. Scanning electron photomicrographs of the polished samples were taken at x100 and randomly assigned identification numbers. The photomicrographs were evaluated and ranked by six dentists for surface smoothness. The best surface polish was obtained with the following sequence: aluminum oxide stone, green Flexie rubber abrasive wheel, white Flexie rubber abrasive wheel, flour of pumice on a wet rag wheel, buffing bar compound on a felt wheel, rouge on a felt wheel, and rouge on a chamois wheel. PMID- 8242015 TI - A review of panoramic radiography and its potential use in implant dentistry. AB - Radiographic follow-up of dental implants is one of the most important clinical parameters a practitioner can assess. Recent advances in the design of panoramic radiograph machines have increased their potential use in the longitudinal clinical evaluation of dental implants. Changes from the earliest designs allow for a projection geometry that more closely approximates the shape of the human jaw. The fundamentals of panoramic radiography are reviewed including common errors in patient positioning, their effect on the radiographic image, and how to correct the errors. Comparative advantages and disadvantages of intraoral periapical and bitewing films as compared with panoramic radiographs are discussed, specifically focusing on the amount of radiation exposure, ability to detect bone loss/bone defects, and inherent problems with both systems. PMID- 8242016 TI - Root intrusion in tooth-implant combination cases. AB - The various proposed etiologies for root intrusion in tooth-implant combination cases are discussed. Clinical examples of root intrusion are illustrated. Advantages and disadvantages of nonrigid connector and coping/suprastructure techniques are presented. PMID- 8242017 TI - Treatment of a patient with multiple implant systems: clinical report. AB - Fabrication of a removable implant-supported prosthesis was complicated by the use of multiple implant systems as described in this clinical report. The management of associated treatment problems is discussed. PMID- 8242018 TI - Flap design considerations in the placement of single maxillary anterior implants: clinical report. AB - Existing flap designs can be modified to improve esthetics in resorbed anterior maxillary edentulous areas when single implants are treatment planned. A clinical report is presented. PMID- 8242019 TI - Transfer of CT scan data to diagnostic casts. AB - A technique for the transfer of diagnostic information from transaxial CT scans to diagnostic casts is presented. A duplicate of the patient's existing prosthesis is utilized as a custom tray and an impression of the edentulous arch is made. The data from the transaxial radiographs are transferred to the diagnostic cast using anatomical landmarks as reference points. The marked cast can be used by the surgeon and the restorative dentist for optimal implant placement. PMID- 8242020 TI - [Rheumatologic manifestations in heart transplant recipients. A cross-sectional study of 365 patients]. AB - Rheumatological complications are sometimes disabling in heart transplant recipients and may negate the good results obtained with transplantation. The objective of this study was to evaluate the incidence of these complications. 365 consecutive heart transplant recipients (292 males and 73 females) were systematically interviewed and examined according to a standardized protocol. The mean age of the patients was 45.9 +/- 12.0 years (range: 11-68). The mean duration from transplantation to time of the study was 35.8 +/- 25.6 months (range: 1-115). The rheumatological disorders most frequently encountered were: gout, osteoporosis, osteonecrosis and myalgias. Early-onset polyarticular gout was diagnosed in 63/365 patients (17.3%). This diagnosis was significantly associated with patient's age, time since transplant, male sex, serum uric acid, serum creatinine, diuretics intake and inversely associated with the serum cyclosporin levels. Hyperuricemia was observed in 75.9% of transplant recipients with a mean of 507.5 +/- 132.5 mumol/l (range: 97-965). An osteoporotic fracture was present in 18/365 patients (4.9%) and was significantly associated with the patient's age, but not with the dose of corticosteroids. Osteonecrosis was detected in 10/365 patients (2.7%), always affected the hip, and was significantly associated with the patient's age, but not with the high doses of steroids. Myalgias were reported by 14/365 patients (3.8%). Laboratory, electromyographic and histological analysis were negative. Rheumatological complications are frequent in heart transplant recipients and justify preventive and therapeutic management. PMID- 8242021 TI - [Sarcoma of the spine in Paget's disease of bone. Apropos of 8 cases]. AB - Eight cases of spinal sarcoma complicating Paget's disease seen in different centers were reviewed. Clinical and radiological features of this condition were determined on the basis of these 8 cases and of 51 previously published cases. In the current series as well as in previous reports, most cases involved the lumbar and sacral spine and manifested as low back pain with sciatica and early development of neurological deficits. Roentgenographic diagnosis proved difficult because of the anatomic complexity of the spine and architectural changes characteristic of the pagetic bone. The most common radiological features was a mixed pattern with both sclerosis and central osteolysis. Computed tomography performed in one patient of current series disclosed osteolysis of the sacrum (not visible on plain roentgenograms) with tumor spread to adjacent soft tissues. Features shared by spinal and nonspinal sarcomas complicating Paget's disease include advanced mean age of patients, predominance of osteogenic sarcomas among histologic forms, increased risk of sarcoma in polyostotic Paget's disease, and a very grim prognosis. Development of a neurological deficit in a patient with Paget's disease is an unusual occurrence which should suggest sarcomatous transformation. Roentgenographic changes are difficult to identify on plain films of the spine and consequently CT scan studies are warranted whenever a sarcoma is suspected. PMID- 8242022 TI - [Extramedullary hematopoiesis. An unusual cause of Pagetic spinal cord compression]. AB - An original case is reported: spinal cord compression due to extramedullary hematopoiesis of the epidural space, related to Paget's disease of bone, successfully treated by surgery. Extramedullary hematopoiesis, usually occurring in cases of hematological disease, can also complicate Paget's disease, probably by bone effraction mechanism. A review of literature has found 5 other cases of extramedullary hematopoiesis occurring in Paget's disease, without hematological disease. Medullar scintigraphy (Indium111, Technetium99 Sulfur colloid) can confirm the diagnosis after X-ray findings, without the need for biopsy. Awareness knowledge of this complication can sometimes avoid invasive investigation. Low dose radiotherapy is useful in the case of hematological disorders. In Paget's disease, there is a risk of sarcomatous degeneration. In our patient, surgery cured cord compression, but it relapsed because of extensive fibrosis, in complicating surgical hemorrhage. PMID- 8242023 TI - [Magnetic resonance imaging in osteoid osteoma]. AB - The purpose of this study was to determine magnetic resonance imaging (MRI) features of osteoid osteomas and to evaluate the place of MRI in the diagnostic workup of suspected osteoid osteoma by comparing its diagnostic value with that of computed tomography (CT). Nine patients with radioclinical features highly suggestive of osteoid osteoma were investigated using bone scintigraphy, CT scan and MRI. MRI sequences were weighted for T1 (TR: 500 ms-TE: 34 ms), T2 (TR:2000 ms-TE:50-100 ms) and T1 after intravenous injection of gadolinium-DTPA contrast material. The appearance of the nidus and surrounding area were analyzed, as well as the accompanying inflammatory reaction involving the bone marrow, soft tissues or synovium. All patients had surgical treatment. Clinical recovery was achieved in every case. Histological studies confirmed the diagnosis of osteoid osteoma in 7 patients. In two cases, histological confirmation was not obtained and the exact nature of the lesion therefore remained in doubt, although clinical and scintigraphic findings as well as the subsequent course were consistent with the diagnosis of osteoid osteoma. In 5 cases, MRI proved inferior to CT scan for detecting the nidus. MRI disclosed edema of the bone marrow not seen on CT scan sections in 5 cases, edema of the soft tissues in three cases, and synovitis with joint effusion in one patient with an epiphyseal osteoid osteoma. At present, MRI plays only an ancillary role in the diagnosis of osteoid osteoma, which rests on the concomitant use of bone scintigraphy and CT scan.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242024 TI - [Prospects of immunotherapy in rheumatoid arthritis]. PMID- 8242025 TI - [Aspergillus spondylodiscitis. Apropos of 5 cases]. AB - Five cases of Aspergillus discitis in male patients are reported. Three patients had impaired immune responses as a result of immunosuppressive therapy following a heart transplant (two cases) or hairy cell leukemia (one case). Two patients had a recent history of mycobacterial infection. All five patients were hospitalized for severe spinal pain suggestive of an inflammatory disease with no neurological abnormalities. Erythrocyte sedimentation rate was elevated in every case. The diagnosis of discitis was suspected on spinal roentgenograms and established by computed tomography and/or magnetic resonance imaging. In three patients the spine was the only site of Aspergillus infection (lumbar discitis in two cases and thoracic discitis in one case). One patient developed Aspergillus infection of several disks (L1-L2, L2-L3, and L4-L5) after Aspergillus endocarditis with embolization to the left lower limb. Another patient developed discitis after an Aspergillus lung infection. In every case, Aspergillus fumigatus was recovered in cultures of specimens harvested by a percutaneous needle biopsy of the intervertebral disk. All five patients were treated by itraconazole which was given as single drug therapy in one case and in combination with 5-flucytosine and amphotericin B in four cases. Recovery was achieved in every case after four to six months of this drug therapy. In contrast to most previously reported cases, none of the five patients reported herein required surgical treatment. Efficacy of conservative treatment in this study may be related to the use of itraconazole in every case.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242026 TI - [Aspergillus spondylodiscitis. Apropos of 3 cases]. AB - Three cases of Aspergillus discitis are reported in immunodepressed patients. Vertebrae osteomyelitis was preceded in 2 cases by pulmonary infection. The diagnosis was established in every case by disc aspiration or closed-needle biopsy and mycological examination of the material. Aspergillus fumigatus was identified in two cases; in one case itroconazole led to recovery within three months. PMID- 8242027 TI - [Pyomyositis. Apropos of 5 cases]. AB - Four patients meeting the "classical" criteria for spontaneous pyomyositis are reported. No local cause was found. Risk factors included diabetes mellitus (2 patients), hemopathy (one patient), and alcohol abuse (one patient). Causative organisms (Staphylococcus aureus in 3 cases and Salmonella sp in one case) were recovered from blood cultures, an unusual occurrence. CT scan studies ensured the diagnosis in every case. An additional case of pyomyositis due to Staphylococcus aureus illustrates the difficulties in the definition of disease. This patient, whose muscle lesions were remarkably well visualized by MRI with injection of gadolinium, developed infection of the sacro-iliac joint adjacent to the muscular focus of infection. This patient may have had either "primary" pyomyositis with spread to the adjacent joint or "secondary" pyomyositis caused by the joint infection which was recognized only later. Advances in medical imaging techniques suggest that the nosology of pyomyositis should be broadened using this terminology. This would underscore the unique characteristics of "classical", "primary" pyomyositis and emphasize imaging, diagnostic and therapeutic facets of the disease which are shared by both entities. PMID- 8242028 TI - [Local factors of bone remodeling]. PMID- 8242029 TI - [Vitamin-resistant hypophosphatemic rickets and spinal cord compression. Apropos of 2 cases]. AB - Adults with X-linked vitamin-resistant hypophosphatemic rickets can develop spinal cord compression as a result of spinal canal stenosis due to involvement of the spine with the increased periosteal and ligamentary calcification which is typical of the disease. Two cases are reported. In both patients, spinal cord compression manifested as rapidly progressive spastic paraplegia due to compression at T9 and T5, respectively. Decompressive laminectomy ensured full recovery in one case but was ineffective in the other. In both cases, paraplegia developed approximately one year after initiation of therapy combining a metabolite of vitamin D and phosphorus in high doses. Current imaging techniques (CT scan, magnetic resonance imaging) provide highly accurate information on the anatomic mechanism of the stenosis, its often multiple locations, and its course. Three factors are involved in the stenosis, in variable degrees: thickening of the laminae, hypertrophy of the facet joints, thickening and calcification of the yellow ligament. Magnetic resonance imaging is the investigation of choice in these patients. A deleterious effect vitamin D-phosphorus treatment cannot be outruled. The appropriateness of initiating this treatment should be prudently discussed and treated patients should be closely monitored. PMID- 8242030 TI - [Cerebral thrombophlebitis disclosing Behcet disease. Value of high resolution MRI]. AB - A 21-year-old male of Moroccan descent was admitted for inflammatory polyarthralgia and monoarthritis of the left knee. The patient reported a history of recurrent isolated aphthous ulcerations of the oral cavity. He was found to be positive for the HLA B5 antigen. The other investigations for Behcet's disease were negative. During hospitalization, headache responsible for insomnia developed. Neurologic evaluation was normal, as well as funduscopy and spinal tap findings. CT scan images were suggestive of cerebral thrombophlebitis. Magnetic resonance imaging (MRI) disclosed an old thrombosis of the right transverse sinus. Time-of-flight MRI (MRI with sequences adapted to the study of vessels) evidenced recent thrombophlebitis of the left transverse sinus. Cerebral angiography fully confirmed MRI findings. Treatment with a corticosteroid and an anticoagulant was successful. Final diagnosis was Behcet's disease. Subsequently, the patient developed additional features of the disease, including hypersensitivity to pinprick, retinal vasculitis, and erythema nodosum. This case is interesting for two reasons: cerebral thrombophlebitis in Behcet's disease can manifest as localized headache. It occurs in 6 to 13% of patients. Early diagnosis and treatment are essential; MRI features of cerebral thrombophlebitis were confirmed by the cerebral angiogram. MRI established that the thrombophlebitis was of recent onset. MRI may be appropriate as the initial investigation in patients with suspected cerebral thrombophlebitis. PMID- 8242031 TI - [Primary bone lymphoma disclosed by osteolytic lesion of the femoral head]. AB - A case of primary lymphoma of the bone is reported. The unusual site of the tumor in an epiphysis (head of femur) led to unconventional therapy by resection and hip replacement. Emphasis is put on the value of immunolabelling studies for the etiologic diagnosis. PMID- 8242032 TI - [Psoriatic arthritis in a patient treated with interferon alpha]. PMID- 8242033 TI - [Subacute development of nephrotic syndrome caused by tiopronin therapy. A propos of 4 cases]. PMID- 8242034 TI - [Tiopronin-induced polymyositis confirmed by histology]. PMID- 8242035 TI - [Anterior pathologic dislocation of the hip in adulthood complicating Von Recklinghausen neurofibromatosis]. PMID- 8242036 TI - Violence and abuse of women: a perinatal health care issue. AB - This article discusses the perinatal health issue of abuse directed at pregnant women by their intimate partners. The incidence, prevalence, and severity of intimate violence during pregnancy are presented, as is a discussion of the possible health disruptions that can occur as a result of violence and abuse. Nursing interventions for abused pregnant women are presented within a framework of advocacy and empowerment. Strategies for primary prevention are outlined, and methods of secondary and tertiary intervention are discussed. The article concludes with a discussion of culturally relevant practice for abused women. PMID- 8242037 TI - Sexual violence and older women. AB - Sexual violence and older women is a growing problem that is underreported in crime statistics and often not identified by clinicians. Overcoming ageism and acknowledging the sexuality of older women are fundamental to the improved recognition of sexual violence throughout the life span. An epidemiologic analysis of factors associated with sexual violence suggests approaches for prevention, improved recognition, and intervention with older women. The use of geriatric assessment parameters, such as functional assessment, is suggested for older female victims of sexual violence, along with physical examination strategies and follow-up interventions, such as counseling and community referrals. Preventive measures include advocacy for safer environments and counseling about potential vulnerability. PMID- 8242038 TI - Sexual assault nurse clinician: an emerging area of nursing expertise. AB - The Sexual Assault Resource Service (SARS), in Minneapolis, Minnesota, has been a leader in developing a new independent role for nurses, the Sexual Assault Nurse Clinician/Examiner. In this role, the nurse works closely with the emergency department staff, the police, and the courts to ensure that proper evidence is collected for use in court to aid in the prosecution of assailants. This program was developed in 1977 when a gap in service delivery to rape victims was identified by obstetric-gynecologic nurses at Hennepin County Medical Center in Minneapolis. An initial 3.5-year demonstration treatment grant allowed development of the model program. The SARS nurse is on call to five community hospitals. When a rape survivor comes to one of the participating hospitals, the SARS nurse on call is paged. In the emergency department, the nurse completes the full evidentiary examination, evaluates the survivor's risk of pregnancy and offers preventive care; treats prophylactically for sexually transmitted diseases; provides crisis intervention; and ensures that injuries are treated by the emergency department staff. The SARS model was developed at the county hospital, but it was expanded to include four local private hospitals. In all hospitals, all services are free of charge to the survivor. There are approximately 12 similar programs throughout the United States. PMID- 8242039 TI - Obstetric-gynecologic care and survivors of childhood sexual abuse. AB - Women's health care providers have few resources to guide them in giving sensitive care to survivors of childhood sexual abuse. This article reviews some commonly seen coping responses, discusses ways in which these coping responses may affect the experience of gynecologic or obstetric care, and provides suggestions on how the care provider can facilitate the patient's process of healing. Also addressed is the need of care providers to process the often intense feelings evoked by issues of sexual violence and abuse. PMID- 8242040 TI - When the patient is also a lesbian. AB - Social stigma impedes the delivery of quality health care to women who identify as lesbians. Nurses who are aware of and sensitive to women who are lesbians in their practice are able to increase the caliber of the health care they provide. This article provides examples in which a lesbian sociosexual orientation has an impact on the medical history, differential diagnosis, physical examination, and treatment. PMID- 8242041 TI - Patterns of alcohol and drug use among women: focus on special populations. AB - Despite the growing national concern about the effect of the use of alcohol and other drugs on the health and productivity of men and women, relatively little is know about alcohol and other drug (AOD) problems in women. Because of this lack of research and the stigma associated with women who are alcoholic or drug addicted, AOD problems among women are less likely to be recognized and addressed. This article reviews patterns and consequences of AOD use among a number of special populations of women. Information about risk factors and assessment and screening are included to assist the nurse in the identification, counseling, and referral of women with AOD-related problems. PMID- 8242042 TI - Women's body image: the nurse's role in promotion of self-acceptance. AB - Women face criticism regarding their bodies from childhood to old age. Overt and covert messages demand that all women seek the ever-changing "ideal" body through a variety of costly, ineffective, and often dangerous means. As these demands are internalized, healthy women may develop a negative body image and a damaged sense of self. Nurses can promote peace between mind and body through the nursing process, role modeling, and community involvement. Body image should be assessed, particularly during vulnerable periods, such as adolescence, pregnancy, postpartum, and menopause. Planning, intervention, and evaluation should focus on self-acceptance and emotional and physical well-being, rather than on changing one's appearance. PMID- 8242043 TI - Women, obesity, and the results of medical management. AB - Women represent more than 75% of the individuals who receive obesity treatment; suprisingly, the numbers of morbidly obese white men and women are approximately equal. Surgical and nonsurgical medical treatments are successful in promoting weight loss; however, maintaining the weight loss is a continuing problem. Weight loss provides many benefits but can lead to undesirable consequences (eg, gallstones and nutrient deficiencies). Long-term success can be accomplished only if the weight loss is permanent; weight cycling increases the health risks associated with obesity. Caution should be exercised in choosing a treatment for obesity so that weight loss is not accomplished at the expense of the health of the individual. PMID- 8242044 TI - Women and eating disorders. AB - The eating disorders of anorexia nervosa and bulimia nervosa are important health problems for women today. Nurses at different practice sites are in a position to assess, treat, and refer these women, depending on educational and clinical preparation. This article presents information to facilitate the role of the nurse in working with women with eating disorders. Clinical features, risk factors, and treatment options are discussed. PMID- 8242045 TI - Interstitial cystitis: women at risk for psychiatric misdiagnosis. AB - Interstitial cystitis (IC) is a form of painful bladder disease, characterized by chronic urinary urgency, frequency, and pain without evidence of bacterial infection. Ninety percent of patients with IC are women. Somatization disorder is a psychiatric diagnosis, given most often to women who report a variety of symptoms, such as dysmenorrhea, burning in sex organs, dyspareunia, irregular menstrual periods, and painful urination. Because diagnosis of interstitial cystitis may take many years, health care providers must be alert to the risks of the overdiagnosis of psychiatric problems and the underdiagnosis of interstitial cystitis. PMID- 8242046 TI - Cardiovascular disease prevention in women and the hormone debate. AB - Because of reported biologic research and mass media attention, the decision to take or not to take noncontraceptive hormones to prevent cardiovascular disease (CVD) is a major women's health issue. This article offers research-based knoweledge on menopause, CVD, and noncontraceptive hormones that is essential for postmenopausal clients and nurses for their thoughtful examination of this issue. By building on this knowledge base and using a client-provider interaction model, a shared decision can be sought that is comfortable and viable for the client. PMID- 8242047 TI - Sexual counseling of women having gynecologic surgical procedures. AB - A feminist, women's health perspective is described to aid nurses in providing sexual counseling to women having gynecologic surgical procedures. Suggestions are given concerning areas of sexual assessment, resource development, and when and what to teach women about the sexual effects of gynecologic operations. PMID- 8242048 TI - Gynecologic issues of women with human immunodeficiency virus infection. AB - Women are the fastest growing population with newly diagnosed acquired immune deficiency syndrome (AIDS) in the United States. Although little is known of specific female manifestations, research trends suggest an increase in genital tract infections and neoplasms that may be recurrent and difficult to treat. The initial presentation of early human immunodeficiency virus (HIV) infection in many women is recurrent and chronic vaginal candidiasis. Abnormal cervical cytology is common, and biannual Papanicolaou (Pap) smear screenings are recommended, preferably with a colposcopic examination. Because such human papillomavirus (HPV)-related dysplasias often are multifocal, anal Pap smears also are recommended. This article addresses the gynecologic problems and care of HIV-positive women. PMID- 8242049 TI - Abortion. AB - Induced legal abortion remains the leading worldwide fertility regulation method. More than 1.5 million abortions are performed in the United States and Canada annually. Nursing is integral to the care of women who have abortions and their families. The complexity of nursing care is dictated by gestational age, the procedure performed, and patient needs. PMID- 8242050 TI - Coronary artery disease in women: assessment, diagnosis, intervention, and strategies for life style change. AB - Coronary heart disease is reaching epidemic proportions in women. With increased longevity, more women are living many years beyond menopause, the time when heart disease peaks in women. For midlife women, it is important to determine a personal profile of unmodifiable (family history, age, and race) and modifiable risk factors. The goal for every woman must be to control coronary risk factors such as smoking, hyperlipidemia, diabetes, sedentary life style/weight control and stress, which have been proven to be amenable to modification strategies. Many women also may consider hormonal replacement but must be counseled to make informed choices concerning the benefits and risks. Finally, as nurses and as leaders in the health care of women, we must model heart healthy behaviors to encourage each other and to increase our effectiveness with the clients we serve. PMID- 8242051 TI - Women and cancer. AB - Cancer will be diagnosed in more than a half a million women this year. Prognosis and curability are associated with the stage of the cancer at diagnosis. If cancer is detected at an early stage, more than 75% of women can expect long-term survival or cure for the most common cancers, excluding lung. Promoting health behaviors that reduce risk and recommending screening tests are critical activities for the practitioner that may affect survival rates and quality of life of women with cancer diagnoses. Risk factors, incidence, presenting symptoms, staging, treatment and 5-year survival rates associated with 12 major cancer sites in women are presented. Diagnosis and survival for minority populations are reviewed, highlighting the need to improve cancer screening and detection for this group of women. PMID- 8242052 TI - Approaches to physical activity in women. AB - It is well known that regular, moderate-intensity exercise is beneficial in promoting good health and preventing disease and disability in women; however, high levels of sedentary behavior are observed in women of all races, incomes, and age groups. The determinants of exercise participation in women are poorly understood. Exercise intervention programs should be designed to increase women's general physical activity habits and enhance physical fitness. Exercise programs should include physical activities that are safe and increase gradually in the frequency, intensity, and duration of effort expended over time. PMID- 8242053 TI - Clinical application of the relaxation response in women's health. AB - Women are subject to multiple stressors and often get little recognition or support for their diverse roles. Although stress is a natural part of life, research indicates that prolonged or extreme stress can have a negative effect on a woman's health. Nurses, because of their holistic approach, are ideally suited to use interventions that empower women to deal effectively with stress. This article outlines the mind/body connection and describes the relaxation response as a counterbalance to the deleterious effects of stress. Instructions for several methods of using the relaxation response in a clinical setting are given. PMID- 8242054 TI - Health hazards of nursing: identifying workplace hazards and reducing risks. AB - Nurses often work in settings in which they may be exposed to a wide array of psychosocial, chemical, biological, and physical hazards. The authors outline several ways in which occupational exposures occur and the general process for reducing or preventing workplace hazards. Several commonly encountered workplace hazards and their potential health risks are identified and discussed. Specific health hazards that are addressed include the chemical hazards of antineoplastic and antiviral drugs; the biological hazards of human immunodeficiency virus, hepatitis B, herpes viruses, rubella, and tuberculosis; and the physical hazards of noise and ionizing and nonionizing radiation. The authors suggest specific preventive measures that nurses can take to make their workplaces safer. PMID- 8242055 TI - Pregnancy and disability. AB - Recently, increasing numbers of women with disabilities are deciding to become pregnant. Several issues pertaining to pregnancy and disability are discussed, and a woman's personal account of her experiences of pregnancy, labor, and delivery after being diagnosed with multiple sclerosis is provided. Specific issues related to her experiences with the health care system are addressed. These include access to health care, self-care practices, sexuality, contraception, prenatal counseling, and parenting. Implications for clinical practice are discussed. PMID- 8242056 TI - A novel SCN4A mutation causing myotonia aggravated by cold and potassium. AB - The single strand conformation polymorphism (SSCP) technique was used to screen genomic DNA of a family with myotonia aggravated by cold, potassium loading and suxamethonium, but without muscle weakness. An aberrant band was found in exon 24 of SCN4A, the gene encoding the adult skeletal muscle sodium channel alpha subunit. DNA sequencing led to the detection of a G-to-A transition of cDNA nucleotide 4765 predicting a substitution of methionine for valine at position 1589 of the protein sequence. This amino acid is located within transmembrane segment S6 of channel repeat IV close to the cytoplasmic surface, a region which is supposed to act as acceptor of the inactivation gate of the channel. Four lines of evidence indicate that this mutation causes the disease: (i) the transition was only found for affected family members; (ii) no mutations were found in all other SCN4A exons; (iii) the affected gene region is conserved among various species; and (iv) an increase in the number of non-inactivating sodium channels had been revealed in earlier electrophysiological studies on an excised muscle specimen from the index patient. In addition, the close-by occurring substitution of valine for methionine at position 1592 known to cause hyperkalemic periodic paralysis was deduced for six families with the myotonic, non-dystrophic form of this disease. PMID- 8242057 TI - Detection of missense mutations by single-strand conformational polymorphism (SSCP) analysis in five dysfunctional variants of coagulation factor VII. AB - Five unrelated subjects with dysfunctional coagulation factor VII (FVII) were studied in order to identify missense mutations affecting function. Exons 2 to 8 and the intron-exon junctions of their FVII genes were amplified from peripheral white blood cell DNA by PCR and screened by SSCP analysis. DNA fragments showing aberrant mobility were sequenced. The following mutations were identified: in case 1 (FVII:C < 1%, FVII:Ag 18%) a heterozygous A to G transition at nucleotide 8915 in exon 6 results in the amino acid substitution Lys-137 to Glu near the C terminus of the FVIIa light chain; in case 2 (FVII:C 7%, FVII:Ag 47%) a heterozygous A to G transition at nucleotide 7834 in exon 5 results in the substitution of Gln-100 by Arg in the second EGF-like domain; in case 3 (FVII:C 20%, FVII:Ag 76%) a homozygous G to A transition at nucleotide position 6055 in exon 4 was detected resulting in substitution of Arg-79 by Gln in the first EGF like domain; in case 5 (FVII:C 10%, FVII:Ag 52%) a heterozygous C to T transition at nucleotide position 6054 in exon 4 also results in the substitution of Arg79, but in this case it is replaced by Trp; case 4 (FVII:C < 1%, FVII:Ag 100%) was homozygous for a previously reported mutation (G to A) at nucleotide position 10715 in exon 8, substituting Gln for Arg at position 304 in the protease domain. Cases 1, 2 and 5 evidently have additional undetected mutations. PMID- 8242058 TI - Cloning of a novel, anonymous gene from a megabase-range YAC and cosmid contig in the neurofibromatosis type 2/meningioma region on human chromosome 22q12. AB - In order to permit detailed characterization of meningioma cases showing deletions within chromosomal band 22q12 and further systematically clone genes located within this region, we established a genomic YAC and cosmid contig which encompasses a region in excess of 1000 kb of 22q12. The YAC contig consists of 6 YAC clones arranged into 5 overlapping steps covering more than 1100 kb. Two corresponding cosmid contigs consisting of 40 steps of overlapping groups of cosmids encompasses 900-1000 kb. This set of genomic clones provides a detailed physical map of this part of chromosome 22 and constitutes a basis for the isolation and characterization of genes that may be located within this chromosomal region. Employing the exon-amplification method on two cosmids from the contig, we cloned a novel, anonymous gene, pK1.3, which potentially encodes a protein of 683 amino acids with a predicted molecular weight of of 78.5 kD. Its 2.7 kb mRNA is expressed ubiquitously. We estimated the genomic size of this gene to 100-150 kb, and it is located in the immediate centromeric vicinity of the neurofibromatosis 2 (NF2) tumor suppressor gene. PMID- 8242059 TI - The human D11S554 locus: four distinct families of repeat pattern alleles at one locus. AB - We have sequenced amplified products of 44 short tandem repeat alleles at the human D11S554 locus. The sequences revealed four distinct, but related, families of consensus repeat patterns. Six alleles deviated from the consensus patterns. Alleles differed in size by as little as one base pair, and precise sizing is required for analysis of alleles at this highly polymorphic locus. The evolution of alleles at the D11S554 locus and related short tandem repeat loci is discussed. PMID- 8242060 TI - Modification of 15q11-q13 DNA methylation imprints in unique Angelman and Prader Willi patients. AB - The clearest example of genomic imprinting in humans comes from studies of the Angelman (AS) and Prader-Willi (PWS) syndromes. Although these are clinically distinct disorders, both typically result from a loss of the same chromosomal region, 15q11-q13. AS usually results from either a maternal deletion of this region, or paternal uniparental disomy (UPD; both chromosomes 15 inherited from the father). PWS results from paternal deletion of 15q11-q13 or maternal UPD of chromosome 15. We have recently described a parent-specific DNA methylation imprint in a gene at the D15S9 locus (new gene symbol, ZNF127), within the 15q11 q13 region, that identifies AS and PWS patients with either a deletion or UPD. Here we describe an AS sibship and three PWS patients in which chromosome 15 rearrangements alter the methylation state at ZNF127, even though this locus is not directly involved in the rearrangement. Parent-specific DNA methylation imprints are also altered at ZNF127 and D15S63 (another locus with a parent specific methylation imprint) in an AS sibship which have no detectable deletion or UPD of chromosome 15. These unique patients may provide insight into the imprinting process that occurs in proximal chromosome 15 in humans. PMID- 8242061 TI - The gene for spinal cerebellar ataxia 1 (SCA1) is flanked by two closely linked highly polymorphic microsatellite loci. AB - The gene for one form of autosomal dominant spinal cerebellar ataxia (SCA1), is mapped by linkage to chromosome 6p, very close to the microsatellite locus D6S89. Eight large Italian kindreds segregating SCA1, as defined by very close linkage to D6S89, were genotyped with five microsatellite markers linked closely to D6S89, all mapping within a 6 cM interval on 6p. Multipoint linkage analysis and haplotypes from recombinants map SCA1 between two of these markers, D6S274 and D6S259, 5-6 cM apart. A single rare four marker haplotype within this interval shows linkage disequilibrium with the disease locus in southern Italy and is transmitted with SCA1 in five kindreds originating from this area. PMID- 8242062 TI - Physical and transcriptional mapping of DXS56-PGK1 1 Mb region: identification of three new transcripts. AB - Several new techniques for isolation expressed sequences have been recently described considerably speeding up the identification of unknown genes. Here we present a transcriptional map of the 1 Mb DXS56-PGK1 region in Xq13.3. Rare cutter restriction site mapping, direct cDNA selection on membrane discs and probing of Northern blots with total YAC DNA, were the methods explored in order to achieve this goal. In addition to three known genes from this region which have been recloned, two new cDNA clones corresponding to two new genes were isolated, mapped and characterized. Moreover one more transcript, highly expressed in placenta, has been detected in the region with a total YAC as a probe. In summary there are at least six genes known to reside in the DXS56-PGK1 region. As several human disease gene loci (i.e. SCID, CMTX1, WWS, MRX, XDP, ASB) were tightly linked to the markers from the region (PGK, CA repeats), the three new transcripts may be considered as their potential candidate genes. PMID- 8242063 TI - Larger expansions of the CTG repeat in muscle compared to lymphocytes from patients with myotonic dystrophy. AB - The discovery that there is an expansion of a CTG repeat underlying myotonic dystrophy has led to new approaches in diagnosis and genetic counselling for this disorder. The size of the expansion correlates to a reasonable degree with the clinical symptoms within a given family. We report comparisons of the length of the expansion seen in lymphocytes and muscle samples from eight patients. In all cases the length of the expansion seen in DNA isolated from muscle was larger than that seen in lymphocytes from the same patient. There was no progression of the expansion over a period of 10-15 years in muscle samples from two of these patients even though there had been significant progression of the severity of symptoms during that time. PMID- 8242064 TI - Haplotype diversity in the human red and green opsin genes: evidence for frequent sequence exchange in exon 3. AB - We studied polymorphisms in the coding sequences of the human red and green opsin genes of 133 Caucasian males. Eleven polymorphic sites were discovered in the red opsin gene, seven of which were in exon 3, three in exon 4 and one in exon 5. Polymorphisms at 8 of these sites resulted in amino acid substitutions which generated a total of 18 unique red opsins in the population. The substitutions at three (S180A, I230T, and A233S) of the 8 sites involve hydroxyl-bearing to non polar amino acid residues, and are therefore likely to alter spectral characteristics of the red pigment. Eight polymorphic sites were observed in the green opsin coding sequences, six of which were in exon 3, one in exon 2 and one in exon 5. Five of the eight involved amino acid substitutions which generated 15 unique green opsins in the population. Substitutions at two of these sites involve hydroxyl-bearing vs. non-polar residues. Six polymorphisms, all of which are located in exon 3, are shared between the red and green opsin genes, essentially making it difficult to assign this exon to either of these genes. Markers in exon 3 are in partial linkage disequilibrium with those in exons 4 and 5, whereas the latter two are in strong linkage disequilibrium with each other. Furthermore, markers in the 5' region of exon 3 are also in only partial (54%) disequilibrium with those in the 3' region. The above results strongly suggest a history of frequent gene conversion, mainly localized to exon 3, in the lineages leading to the human red and green opsin genes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242065 TI - Severe childhood autosomal recessive muscular dystrophy with the deficiency of the 50 kDa dystrophin-associated glycoprotein maps to chromosome 13q12. AB - We have recently demonstrated the specific deficiency for the 50 kDa dystrophin associated glycoprotein (50DAG) in Algerian patients afflicted with severe childhood autosomal recessive muscular dystrophy with DMD-like phenotype (SCARMD). A similar disease affecting Tunisian patients was linked to chromosome 13q but the status of the 50DAG was not investigated. Here we show by linkage analysis of Algerian families that the genetic defect which leads, either directly or indirectly, to the deficiency of the 50DAG in skeletal muscle is localized to the proximal part of chromosome 13q. We have not found any evidence of genetic heterogeneity among the thirteen families studied. It remains to be demonstrated whether the 50DAG gene maps at 13q12, and to determine if it is mutated in this disease. PMID- 8242066 TI - Fragile X syndrome unstable element, p(CCG)n, and other simple tandem repeat sequences are binding sites for specific nuclear proteins. AB - The trinucleotide repeat sequences which become unstable in fragile X syndrome and myotonic dystrophy are located in the untranslated regions of their respective genes, FMR1 and DM1. This implies that a functional constraint other than coding capacity maintains the presence of the repeats. In the case of fragile X syndrome, sequences adjacent to the repeat are methylated in affected individuals and the FMR1 gene is transcriptionally inactive. We demonstrate that the fragile X p(CCG)n repeat itself is methylated in vivo and that methylation of this repeat is able to inhibit in vitro binding of a novel, specific nuclear p(CCG)n binding protein (CCG-BP1)--one of at least 10 distinct simple tandem repeat sequence binding proteins (STR-BPs). We describe additional, apparently distinct, binding activities both for the methylated form of the p(CCG)n repeat and for each of the single strands of the repeat. PMID- 8242067 TI - Substitution of Leu for Pro-193 in the insulin receptor in a patient with a genetic form of severe insulin resistance. AB - Mutations have been identified in the insulin receptor (IR) gene in patients who are insensitive to insulin action. We studied an extremely insulin resistant patient whose insulin binding to Epstein-Barr virus (EBV) transformed lymphocytes was severely reduced. Transmembrane signalling, evaluated as insulin receptor autophosphorylation, was normal. The patient's IR was immunoprecipitated normally by AbP6, a polyclonal antibody directed to the beta subunit. However, there was an approximately 50% decrease in the affinity of IR immunoprecipitation by a monoclonal antibody (MA-10) directed against the alpha subunit. These observations suggested that there were likely to be a mutation in the patient's insulin receptor that caused misfolding of the IR alpha subunit. Analysis of gene structure by Southern blotting experiments did not reveal any major deletion in the IR gene of the proband. Northern blot analysis showed a normal level of expression of IR gene. We applied denaturing gradient gel electrophoresis (DGGE) as well as direct sequence analysis to study the 22 exons of IR gene amplified by polymerase chain reaction (PCR) using the proband's genomic DNA as a template. We identified a new missense mutation substituting leucine (CTG) for proline (CCG) in homozygous state at codon 193 in exon 3. Both parents are heterozygous for the Leu193 mutation. The Leu193 mutation was not detected in any of 75 normal subjects (150 chromosomes), indicating that it is not a common sequence variant of the insulin receptor. In addition, during the course of screening the patient's DNA with perpendicular DGGE, we identified two previously unreported silent substitutions in exon 9.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242068 TI - The density of transcriptional elements in promoter and non-promoter sequences. AB - Using data currently available from transcriptional element (TE) databases, we have analyzed the density of these elements in both promoter and non-promoter sequences obtained from the GenBank sequence database. The density of putative TEs in non-promoter sequences was about 16% higher than that seen in pseudo random DNA sequences. Promoter TE density from the transcription startsite to 100 basepairs upstream was found to be about 42% higher than in non-promoter sequences. However, the extensive overlap of putative TE densities between promoters and non-promoters confounds an attempt to use TE density as a simple discriminator. PMID- 8242069 TI - Correct usage of a mutated G+1 splice site and transcript quantitation in a lipoprotein lipase-deficient patient. AB - The consequence on the splice mechanism of a mutation occurring at the donor splice site of intron 1 has been studied. We demonstrate that, in spite of the change at G+1 position, this site is still utilized and can produce correctly spliced transcript. Nevertheless the mRNA is detectable only after an 'in vitro' amplification. A procedure has been developed to reveal and quantify the minute amount present in the patient. The very low mRNA level results in a total lack of enzyme, the cause of the disease. The procedure can be useful in cases of rare transcripts and when the source is limited. Furthermore we analyse the interaction between the splice consensus sequence and the small nuclear RNA, that is the necessary intermediate of the splicing mechanism. We speculate on the reasons why cryptic sites are not utilized and only the authentic site can be used, although significantly destabilized by the mutation. PMID- 8242070 TI - Localization of the human TAX-1 gene to 1q32.1: a region implicated in microcephaly and Van der Woude syndrome. PMID- 8242071 TI - Somatic mutations of the APC gene in precancerous lesion of the stomach. AB - The adenomatous polyposis coli (APC) gene, responsible for familial adenomatous polyposis, is also associated with development of sporadic tumors in digestive system as colon, stomach, or pancreas. In order to investigate whether or not APC mutations occur as an early genetic event during gastric carcinogenesis, we examined somatic mutations of APC in flat adenomas of the stomach. DNAs isolated from flat adenomas were examined by means of an RNase protection analysis coupled with polymerase chain reaction (PCR) followed by DNA sequencing of the PCR products. By screening a mutation cluster region (MCR: codons between 1286 and 1513) of APC in which two-thirds of somatic mutations were detected in colorectal tumors, somatic mutations were found in four of ten flat adenomas: three of which caused truncation of the gene product due to a nonsense mutation or 4-bp deletion; one other was a point mutation that altered amino acid from alanine to threonine. Our results imply that APC plays a crucial role in an early step of gastric carcinogenesis, as was observed in colorectal carcinogenesis. PMID- 8242072 TI - Expansion of the (CAG)n repeat causing Huntington's disease in 352 patients of German origin. PMID- 8242073 TI - Multiple polymorphisms within the alpha-L-iduronidase gene (IDUA): implications for a role in modification of MPS-I disease phenotype. PMID- 8242074 TI - Trinucleotide repeat elongation in the Huntingtin gene in Huntington disease patients from 71 Danish families. AB - IT15 is a novel gene, localized to chromosome 4, and encoding a protein named Huntingtin. A polymorphic CAG repeat in the proposed open reading frame of IT15 has been characterized, and an elongation of this repeat has been correlated to Huntington's Disease. We have investigated the CAG repeat in the Huntingtin gene in 71 unrelated Danish patients with Huntington's Disease, and found repeat lengths of 39 to 70 repeat units in contrast to 9 to 30 CAG's on normal chromosomes. Comparison of repeat length and age at onset of disease symptoms in 52 individuals indicates an inverse correlation between the age at onset and the number of CAG repeat units. PMID- 8242075 TI - Endothelin 1 is not a candidate gene for spinal cerebellar ataxia 1. PMID- 8242076 TI - Single-step screening method for the most common mutations in familial adenomatous polyposis. PMID- 8242077 TI - Genetic heterogeneity of autosomal dominant cerebellar ataxia type I: evidence for the existence of a third locus. PMID- 8242078 TI - A new (old) deletion in the choroideremia gene. PMID- 8242079 TI - Two novel mutations: 5108delAG and 5816insG in the NF1 gene detected by SSCP analysis. PMID- 8242080 TI - A homozygous 5 base-pair deletion in exon 10 of the adenosine deaminase (ADA) gene in a child with severe combined immunodeficiency and very low levels of ADA mRNA and protein. PMID- 8242081 TI - Identification of a single base pair deletion (40 del G) in exon 1 of the ferrochelatase gene in patients with erythropoietic protoporphyria. PMID- 8242082 TI - Four STR polymorphisms map to a 500 kb region between DXS15 and DXS134. PMID- 8242083 TI - Five dinucleotide repeat polymorphisms on human chromosome 16q24.2-q24.3. PMID- 8242084 TI - Six dinucleotide repeat polymorphisms on human chromosome 16q12.1-q24.1. PMID- 8242085 TI - Three dinucleotide repeat polymorphisms on human chromosome 16p13.11-p13.3. PMID- 8242086 TI - Two simple repeat polymorphisms at DXS337. PMID- 8242087 TI - Dinucleotide repeat polymorphism in the interferon-gamma (IFNG) gene. PMID- 8242088 TI - Trinucleotide repeat polymorphism at DXS101. PMID- 8242089 TI - Primers for the dinucleotide repeat at the DXS453 locus also recognizes the DXS983 locus. PMID- 8242090 TI - Dinucleotide repeat polymorphism in the promoter region of neurotrophin-3 gene (NT3). PMID- 8242091 TI - Trinucleotide repeat polymorphism at D6S366. PMID- 8242092 TI - Dinucleotide repeat polymorphism at the KCNA5 locus. PMID- 8242093 TI - Tetranucleotide repeat polymorphism at the D8S320 locus. PMID- 8242094 TI - New human DNA polymorphisms submitted to the genome data base. PMID- 8242095 TI - Improved PCR conditions for the stretch of (CAG)n repeats causing Huntington's disease. PMID- 8242096 TI - Synthesis and secretion of human serum albumin by mammary gland explants of virgin and lactating transgenic mice. AB - Transgenic mice were produced, carrying hybrid genes comprised of the ovine beta lactoglobulin (BLG) milk protein gene promoter and human serum albumin (HSA) coding sequences. In situ hybridization revealed high levels of BLG/HSA hybrid mRNA, confined to the epithelial cells of the lactating mammary gland with a several hundred fold lower concentration in virgin mammary glands. During the first 24 h in culture, exceptionally high levels of HSA were secreted from explants of virgin mice, independent of hormonal control. HSA secretion was reduced considerably during subsequent days in culture and became dependent on the presence of insulin, hydrocortisone and prolactin. This temporal and hormonal pattern of regulation of HSA was different than that found for the secretion of caseins. In contrast to the vast difference in the mRNA content, the amount of HSA secreted from explants derived from lactating mice during the first 24 h in culture was only 2- to 5-fold higher than that found with explants from virgin transgenic mice, suggesting post-transcriptional control of HSA synthesis. The high-level synthesis and secretion of HSA in mammary explants of lactating mice was also dependent on the presence of insulin, hydrocortisone and prolactin. This study confirms previous suggestion that mammary explants from virgin transgenics may serve as a powerful tool for screening the potential of transgenic animals to secrete foreign proteins in their milk. PMID- 8242097 TI - Transgenic animals as models for human disease--report of an EC Study Group. PMID- 8242098 TI - [Positive serology for syphilis in neonatal period: prevalence in secondary level maternity. Association with maternal risk factors and with positive serology for HIV-1]. AB - To study neonatal Syphilis seropositivity incidence, its association with HIV-1 seropositivity and the impact of Congenital Syphilis CDC case definition, cord blood VDRL and Indirect Hemagglutination (IHA) for Syphilis were performed in 3,664 newborns from Dec/91-July/92. A positive group was formed by those with positive VDRL and IHA or just positive IHA. The control group consisted of 200 VDRL and IHA negative neonates. Among the 3,664 neonates 5.6% were seropositive for Syphilis. A significant association was noted (p < or = 0.05) between neonatal seropositivity for Syphilis and mothers older than 20 years, single, with sexual promiscuity, drug use, no pre-natal care, multiparity and previous still-births. All positive neonates fitted in the Presumptive Congenital Syphilis CDC definition. In 50 seropositive for Syphilis neonates ELISA for HIV-1 was performed and 6/50 were positive. Congenital Syphilis is of concern, specially in neonates of mothers with risk factors for Syphilis acquisition. HIV-1 should be considered in seropositive for Syphilis neonates. Adoption of Congenital Syphilis new CDC case definition is advised, particularly in countries where prenatal care is deficient. PMID- 8242099 TI - [Acute hydrothorax as a complication of peritoneal dialysis: report of a case and review of the literature]. AB - Report a case of a patient suffering from acute pancreatitis with renal failure, who needed peritoneal dialysis and developed acute hydrothorax as a complication of the procedure. The diagnosis was made by the high levels of glucose on pleural effusion (455 mg/dL) and glycemia of 81 mg/dL). This complication probably occurs because of a pathological diaphragmatic defect, communicating peritoneal and pleural cavities. The treatment consists of prompt interruption of peritoneal dialysis with improvement of symptomatology. PMID- 8242100 TI - [Tumor necrosis factor: biology, immunology, and systemic impact]. PMID- 8242101 TI - [Arthrosis and headache]. PMID- 8242102 TI - [Style of technologic and scientific publications]. PMID- 8242103 TI - [Coronary angioplasty and revascularization surgery: partners or opponents?]. PMID- 8242104 TI - [Acute myocardial infarction in the thrombolytic era: high mortality in elderly patients]. AB - Patients with acute myocardial infarction (AMI) admitted to a general hospital are very different from those reported in large multicenter trials. PURPOSE: To evaluate the in-hospital mortality in patients with AMI admitted to a general hospital in the thrombolytic era, and to identify factors associated with poor prognosis. METHODS: 113 consecutive patients admitted with AMI in one year at the Hospital de Clinicas of Porto Alegre were studied. Several clinical and laboratory variables related to higher risk were evaluated. According to the hospital routine, 47% of the patients received thrombolytic therapy, and the effects and complications of its use were studied. RESULTS: In-hospital mortality was 19.5%. By univariate analysis, age, female gender and peak creatinephosphokinase level were significantly associated with early mortality. By multiple logistic regression analysis, advanced age was the only predictive variable, even after adjusting for other baseline risk factors. In patients older than 70 years, in-hospital mortality was 41% versus only 8.1% in the younger group. There was no difference in survival with the presence of others factors such as previous myocardial infarction, use of thrombolytic therapy, heart failure, infarcted area, diabetes, and systemic hypertension. Among patients who received thrombolytic therapy, an elevated number of adverse effects was observed, and in 9.4% of the cases the complications were serious. CONCLUSION: The high in-hospital mortality after AMI in a general hospital is associated with the patients' advanced age. In spite of recent therapeutic advances in the treatment of AMI, older patients have not obtained a comparable benefit as that achieved by the younger group. PMID- 8242105 TI - [A psychosomatic approach to perennial allergic rhinitis]. AB - The objective of this investigation is to study the involvement of the psychological process in the etiology of perennial allergic rhinitis. Thirty patients with perennial allergic rhinitis, submitted to a Program of Hyposensibilization to Home Dust at the ENT Department, were selected. All patients were studied through psychological interviews and Pfister Colored Pyramid Test. Our results were compared to those obtained by Robert Heiss in his studies of standardization parameters (non rhinitic sample). The study revealed that our patients present intrinsic psychological difficulties characteristic of failures in the affective--emotional development and points out to a neurotic line of personality structuration. PMID- 8242106 TI - [Ventricular function in animals undergoing renal mass ablation. Effect of blood pressure control]. AB - The causes of left ventricular hypertrophy (LVH) in chronic renal failure are multifactorial. The tail arterial pressure, heart weight, maximum developed pressure and pressure-volume relationship in animals with 5/6 nephrectomy (CRF, n = 16) and CRF animals who received furosemide by oral gavage (CRF-F, n = 17) were studied. All CRF animals increased their blood pressure (Basal: 117.2 +/- 0.8; 6th week: 184.8 +/- 2.9, p < 0.001) but in those treated with furosemide the blood pressure stayed within normal levels (Basal: 118.23 +/- 1.3; 6th week = 118.8 +/- 1.6, p = 0.5734). SHAM animals served as control (SHAM, n = 15). All CRF and CRF-F animals increased their levels of serum creatinine (CRF = 1.81 +/- 0.05; CRF-F = 1.75 +/- 0.05; SHAM = 0.72 +/- 0.09 mg/dL p < 0.001). Cardiac weight was elevated in CRF and CRF-F when compared with SHAM rats. The operational maximum developed pressure was similar in the three groups (CRF = 221.2 +/- 7.04; CRF-F = 255.7 +/- 10.1; SHAM = 239.9 +/- 5.5 mmHg, n.s.). However, the end-diastolic volume was significantly increased in both CRF and CRF F when compared with SHAM rats (CRF = 91.4 +/- 9.44; CRF-F = 70.9 +/- 6.22; SHAM = 28.75 +/- 3.12 microL. p < 0.001). These data demonstrate that LVH in chronic renal failure is not dependent of the arterial hypertension, that accompanies this condition. Moreover, the systolic work of LVH animals is similar to that of normal animals, but a greater utilization of Frank-Starling mechanism to maintain normal ventricular function is needed. PMID- 8242107 TI - [Prevalence of hypercalcemia in patients with lung cancer]. AB - Although hypercalcemia is a serious and frequent complication of lung cancer, it is not commonly investigated in our country. We studied the prevalence of hypercalcemia in 90 random lung cancer patients from the Department of Pneumology, Escola Paulista de Medicina. The following histological types were found: 35 Squamous cells carcinomas (CEC), 30 Adenocarcinomas (AdenoCa), 11 Small cells carcinomas (CIPC), 2 Large cells carcinomas (TGC), 1 Carcinoid, 1 Mesothelioma, 2 Undifferentiated carcinomas, 1 Adenosquamous, 1 in situ carcinoma and 4 metastatic tumors. Ionized Ca (Ca-i) was measured in blood samples of all patients. Hyperparathyroidism was excluded by PTH and cAMP determinations in the hypercalcemic patients (Ca-i > 1.29 mmol/L). We found elevated levels of Ca-i (range = 1.30 to 2.0 mmol/L) in 18 patients (20%), being: 12 CEC (66.7%), 3 AdenoCa (16.7%), 2 CIPC (11.1%) and 1 TGC (5.6%). The PTH levels were low or suppressed in all 18 patients, but cAMP determinations were elevated in 6 out of 12 patients. Hypercalcemia is then a very frequent complication of lung cancer (20%), and PTH measurement was able to exclude a hyperparathyroidism in all cases studied. PMID- 8242108 TI - [Complications of central venous catheters: prospective study]. AB - The insertion of central venous catheters (CVC) represent a very common procedure in intensive care units. The purpose of this study was to determine the via that provide the best chance for a good location of the tip of a CVC. The 563 CVCs that were inserted percutaneously and that had a radiology control were analyzed. The infraclavicular via was used for 303 (25%) CVCs; the jugular via had 142 (25%) CVCs and the supraclavicular via was used in 118 (21%) CVCs. The rate of bad location of the tip of the CVC were 17%, 6% and 9%, respectively. There was a significant difference when the insertion through infraclavicular via and jugular via were compared. There were 9 pneumothorax (1.4%). 127 CVCs were sent to microbiology laboratory, 26 (20%) had positive culture (> 15 colony forming units). The most frequent agents isolates were: Candida spp (7), S. aureus (5), S. epidermidis (4) and P. aeruginosa (3). PMID- 8242109 TI - [A short-term randomized controlled study with methotrexate in rheumatoid arthritis]. AB - Randomized, controlled and double-blind study of 36 patients aimed at the evaluation of the efficacy and toxicity of MTX in the treatment of rheumatoid arthritis. Twenty-eight patients completed the study period: 14 in the MTX group and 14 in the placebo group. The patients treated with MTX presented a statistically significant improvement (p < 0.05) in pain, grip strength and functional ability when compared to placebo treated patients. Mild adverse effects were observed in 4 patients treated with MTX and in 2 patients treated with placebo. These findings support other studies and give to methotrexate a relevant position in the treatment of rheumatoid arthritis, owing to its convenient posology, beneficial effectivity and favourable toxicity. PMID- 8242110 TI - [Professional profile of students in specialization courses on Occupational Medicine]. AB - The authors tied to find out a physician's professional profile who attended specialization courses in Occupational Medicine offered by the University of Sao Paulo Medical School. There are eight charts that show how the courses developed and the necessary changes made for their recognition in this specialty. The results also show how many jobs a physician needs to obtain a reasonable salary for his survival. PMID- 8242111 TI - Should federal health care reform be financed by an employer mandate? No. PMID- 8242112 TI - Should federal health care reform be financed by an employer mandate? Yes. PMID- 8242113 TI - Will the Clinton plan help or hinder integration? PMID- 8242114 TI - Finding your way. Networks need more enforcement guidance on a tangle of legal issues. AB - Health care attorneys say more guidance is needed for their clients as they structure integrated health networks, so they can avoid violating laws governing fraud and abuse, physician referrals, competitiveness and tax-exempt status. Unfortunately, for the time being, there's still relatively scant advice federal agencies can give hospitals, physician groups and other provider organizations in these areas. An overview of the issues, and views from regulators and experts. PMID- 8242115 TI - The measuring stick. Is health care ready for full-scale outcomes measurement? PMID- 8242116 TI - Bridging the gap. Home-and community-based programs integrate acute and long-term care. PMID- 8242117 TI - The quality march. National survey profiles quality improvement activities. AB - This nationwide profile of CQI/TQM adopters and non-adopters provides important baseline information with which to chart the growing involvement of hospitals with formal quality improvement efforts. Using a stringent definition, the findings suggest rather widespread adoption of CQI/TQM, although most of it has been very recent. Further, there are systematic differences by bed size, teaching orientation, and system membership. Though the Deming method is the most popular approach to CQI/TQM, nearly as many hospitals report using a combination of approaches, and approximately 22 percent report that they have not selected any specific approach. Of particular note is the finding that those involved with CQI/TQM activities perceive fewer barriers to their quality improvement efforts than those not involved. The impact of these differences on perceived costs and outcomes will be addressed in the next issue of Hospitals & Health Networks. PMID- 8242118 TI - Miami, FL. As physician/hospital organizations make a splash in Miami, ripples are being felt up the coast. PMID- 8242119 TI - GHAA President Karen M. Ignagni. Interview by Frank Cerne. PMID- 8242120 TI - Data watch. Community hospitals: expenses leveling off. PMID- 8242121 TI - Public hospitals gain--and lose--under Clinton reform plan. PMID- 8242122 TI - Hospitals put supply contracts under microscope. PMID- 8242123 TI - Some further thoughts on the reform process. PMID- 8242124 TI - Top 10 reasons why many PHOs are failing and hospitals would have been better off doing nothing. PMID- 8242125 TI - Techniques for the measurement of human energy expenditure: a practical guide. AB - We have tried to provide a brief guide to the range of techniques available for assessing energy expenditure in a variety of different settings. It is hoped that this will enable potential users to select an appropriate method for the problem in hand and to be able to trace more detailed reference material, advice or collaboration. The Appendix provides a selection of equipment suppliers known to us. It is not an exhaustive list. Many laboratories which possess some of the more elite techniques are prepared to participate in collaborative studies if approached with a sufficiently novel hypothesis or clinical problem. Since many biological questions will only be solved by harnessing the combined power of several techniques (e.g. whole-body calorimetry and DLW), such collaborative studies are often beneficial. PMID- 8242126 TI - Improvement in arterial stiffness during hypolipidaemic therapy is offset by weight gain. AB - Fourteen patients with familial hypercholesterolaemia were managed with dietary advice and simvastatin for 12 months. Either nicotinic acid or cholestyramine resin was added to the regimen if serum cholesterol was not less than 5.5 mmol/l within 18 weeks. After dietary advice but before commencing pharmacotherapy for hyperlipidaemia, arterial stiffness was measured in the common carotid and common femoral arteries. These studies were repeated after 12 months on pharmacotherapy. The primary objective of this study was to determine whether arterial stiffness could be altered with total cholesterol and low density lipoprotein (LDL) cholesterol lowering. Over the 12 month interval, serum total cholesterol, LDL cholesterol and triglycerides fell significantly, whereas high density lipoprotein (HDL) cholesterol and body mass index (BMI) rose significantly. Mean supine blood pressure did not change significantly. Arterial stiffness in the common carotid artery decreased from 1.04 +/- 0.21 x 10(5) N/m2 to 0.63 +/- 0.06 x 10(5) N/m2 (T = -2.67, P < 0.01) over the interval. Stiffness of the common femoral artery decreased from 2.10 +/- 0.57 x 10(5) N/m2 to 0.83 +/- 0.15 x 10(5) N/m2 (T = -2.73, P < 0.01). The change in arterial stiffness was not directly related to changes in circulating lipids or supine blood pressure. Increase in BMI, however, correlated with change in arterial stiffness in the common femoral artery (Rs = 0.53, P < 0.05) but not in the common carotid artery. An increase in BMI was associated with a smaller decrease in common femoral arterial stiffness. Aggressive hypolipidaemic therapy was therefore associated with a favourable effect on arterial wall stiffness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242127 TI - The relationship between body mass and cigarette smoking using a biochemical index of smoking exposure. AB - This study evaluated the relationship between smoking and body mass using objective indices of both smoking exposure (COHb) and body mass. The subjects were 4240 adults who participated in the Second National Health and Nutrition Examination Survey (NHANES II). Measurements of dietary intake, physical activity, demographics, body mass index (BMI) and blood carboxy-haemoglobin (COHb) were used in these analyses. After controlling for covariables of body mass, the results indicated that smokers displayed a significantly lower mass compared with non-smokers. Smokers with higher levels of COHb had lower BMIs than smokers at lower COHb levels. White smokers had lower BMIs with increasing COHb exposure whereas black smokers had BMIs at high COHb exposure similar to those of non-smokers. Results indicated that smokers weighed less than non-smokers and that the weight control 'benefits' of smoking were most pronounced in white subjects who were heavy smokers. PMID- 8242128 TI - Beta-endorphin response to exogenous corticotrophin-releasing hormone in obese women with different patterns of body fat distribution. AB - Obese individuals may be characterized by higher than normal basal and stimulated beta-endorphin plasma concentrations, which suggests an increased activity of the opioid system. This study was carried out to investigate whether the regulation of beta-endorphin secretion may be different in different phenotypes of obesity. Twenty-two obese women (body mass index greater than 30 kg/m2) without other endocrine and metabolic abnormalities were investigated. A group of seven normal weight healthy women matched for age served as controls. According to the protocol, obese women included in the study had a waist-to-hip ratio higher than 0.85 (n = 9) or lower than 0.80 (n = 13). The former were defined as having abdominal type and the latter peripheral type body fat distribution. Both groups were matched for body mass index. All women randomly underwent a corticotrophin releasing hormone test (human CRF, 1 microgram/kg body weight) and a control saline study, with blood samples for beta-endorphin determination taken at regular intervals. Basal beta-endorphin levels were not significantly different between the three groups. No significant variation in the hormone levels occurred during the control study in either group. After CRF injection, however, beta endorphin rose significantly in all women, but the hormone concentrations were significantly higher in obese women with abdominal fat distribution than in those with peripheral fat distribution and in controls. These results indicate that, among obese women, only those with the abdominal phenotype seem to have increased opioid activity. PMID- 8242129 TI - Effects of exercise and stress on body fat distribution in male cynomolgus monkeys. AB - The effects of exercise and stress on regional and whole body adiposity were examined in an established animal model of diet-induced coronary artery atherosclerosis, the cynomolgus monkey (Macaca fascicularis). A total of 79 adult male monkeys were assigned to four experimental groups after baseline stabilization and training: (i) exercise, stress, (n = 20); (ii) exercise, no stress (n = 20); (iii) sedentary, stress (n = 20); and (iv) sedentary, no stress (n = 19). The monkeys consumed an ad libitum diet containing 188 mg cholesterol per day with 43% of calories as saturated fat. Anthropometric measurements of regional and whole body adiposity were collected throughout the study. A subset (n = 40) of animals representing all four groups underwent computerized tomography (CT) scans at the end of the study to determine amounts of total abdominal, intra-abdominal and subcutaneous abdominal adipose tissue. Results indicate that, in general, stress interacted with exercise to affect anthropometric measurements of regional adiposity. In contrast, stress had independent and significant effects on the amount and distribution of abdominal fat as measured using CT. Stressed monkeys in both the exercise and sedentary groups had more intra-abdominal fat (and thus greater intra-abdominal :subcutaneous abdominal fat ratios) than their nonstressed counterparts. There were no significant interactions between exercise and stress or exercise effects on abdominal fat distribution as measured by CT. These results support the belief that an arousal syndrome caused by chronic stress, and resulting in increased activity along the hypothalamo-adrenal axis, may play a role in the preferential deposition of fat in the abdomen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242130 TI - Comparison of obese and non-obese patients with gestational diabetes. AB - The aim of this study was to assess the effect of obesity on the outcome of gestational diabetes (GD). The age of the subject (31 +/- 5 years) and duration of pregnancy (29 +/- 5 weeks) were similar at the time of diagnosis for the 19 obese (34 +/- 6 kg/m2) and 19 non-obese (23.0 +/- 2.1 kg/m2) subjects. Before treatment, the area under the curve of the glucose tolerance test and the mean capillary blood glucose levels were similar in both groups; however, the baseline blood glucose levels of the obese patients (6.0 +/- 0.8 mmol/l) were higher than those of the non-obese patients (5.3 +/- 0.8 mmol/l; P < 0.01). During pregnancy, the total weight gain of the obese patients was less (10.6 +/- 5.0 vs. 15.2 +/- 4.7 kg, P < 0.006). Fifteen of the obese patients required treatment with insulin, while only six of the non-obese women required insulin (P < 0.005). During insulin therapy, the mean capillary blood glucose, glycosylated haemoglobin and fructosamine levels were similar in both groups. In conclusion, our data suggest that GD in obese patients is characterized by lower weight gain and higher baseline glucose with the result that insulin therapy is more frequently instituted. PMID- 8242131 TI - Increased appetite after prenatal haloperidol exposure--a case report. PMID- 8242132 TI - A brief history of oligonucleotide synthesis. PMID- 8242133 TI - Synthesis of 2'-O-alkyloligoribonucleotides. PMID- 8242134 TI - An improved method for the synthesis and deprotection of methylphosphonate oligonucleotides. AB - The methylphosphonate oligonucleotide synthesis methods described here give the desired products in good yield. Superior amounts of product are achieved by modifying both the DNA synthesis program and the reagent to compensate for the unstable methylphosphonite intermediate. Deprotection conditions have also been altered to maximize the recovery of oligonucleotide from DNA synthesis supports and to minimize the amount of base modification. Mass-spectrometry analysis of our oligonucleotides has verified their purity and confirmed the absence of modified bases. When compared to standard DNA synthesis methods, this procedure uses only about one-third the usual amount of monomer. Using these procedures, it should be possible to synthesize reliably methylphosphonate oligonucleotides at 1 and 15-mumol scales. PMID- 8242135 TI - Oligonucleoside phosphorothioates. PMID- 8242136 TI - Oligonucleotide synthesis. Phosphotriester approach. PMID- 8242137 TI - Synthesis and purification of phosphorodithioate DNA. PMID- 8242138 TI - Oligodeoxyribonucleotide phosphotriesters. PMID- 8242139 TI - Oligonucleoside boranophosphate (borane phosphonate). PMID- 8242140 TI - Oligonucleotide phosphorofluoridates and fluoridites. PMID- 8242141 TI - alpha-Oligodeoxynucleotides. PMID- 8242142 TI - Stereospecific synthesis of P-chiral analogs of oligonucleotides. PMID- 8242143 TI - Oligonucleotide analogs with dimethylenesulfide, -sulfoxide, and -sulfone groups replacing phosphodiester linkages. AB - Two sets of experimental protocols are given for the synthesis of 3',5'-bis homodeoxyribonucleosides, building blocks for the synthesis of oligodeoxynucleotide analogs where the -O-PO2-O- groups are replaced by -CH2-S CH2-, -CH2-SO-CH2-, and -CH2-SO2-CH2- units. Conditions are presented for joining these building blocks to create short nucleic acid analogs. Since isosteric, achiral, and non-ionic analogs of natural oligonucleotides stable to both enzymatic and chemical hydrolysis, such molecules have potential application as probes in the laboratory, in studies of the biological function of individual genes, and as "antisense" oligonucleotide analogs. PMID- 8242144 TI - Oligodeoxyribonucleotides synthesis. Phosphoramidite approach. PMID- 8242145 TI - Scale-up of oligonucleotide synthesis. Solution phase. PMID- 8242146 TI - Large-scale oligonucleotide synthesis using the solid-phase approach. PMID- 8242147 TI - Solid-phase supports for oligonucleotide synthesis. PMID- 8242148 TI - Oligodeoxynucleotide synthesis. H-phosphonate approach. PMID- 8242149 TI - Oligoribonucleotide synthesis. The silyl-phosphoramidite method. PMID- 8242150 TI - Organization development: a role for your second-generation clinical nurse specialist. AB - The demands of today's busy health care environment, coupled with economic constraints, are challenging health organizations more than ever before. The quality of the care we deliver will be jeopardized unless we shift to a more proactive approach, in which we create change, rather than react to it. The systems that will prevail and the individuals who make up those systems will develop the necessary skills, resources, and psychological stamina to succeed in the future. This will be accomplished through attention to people, for it is through people that all change occurs. Second-generation CNSs with advanced skills in process consultation, systems diagnosis, and human relations can assist in the achievement of these goals. The success of the CNS and the benefits to the nursing organization may be strongly influenced by nurse executives who embrace the concept of organization development, choose a skilled CNS for this purpose, and adequately prepare the organization for the utilization of this invaluable resource. PMID- 8242151 TI - Evaluating instructional software. AB - The purchase of educational software for use in the hospital is a major decision. A careful evaluation will prevent the purchase of an expensive program that is not used. There are many questions to be considered, but there is also a great deal of help available. PMID- 8242152 TI - Economic trends in health care. PMID- 8242153 TI - [The development of maternal-fetal attachment during pregnancy]. AB - This study is to examine the degree to which women engage in attachment behaviors toward their unborn children. Cranley's Maternal-Fetal Attachment Scale (MFAS) was modified to Japanese version (MFAS-J2), which consists of 20 items with .87 reliability. MFAS-J2 was administered to both normal and high-risk pregnant women (n = 275) during gestation. All subjects were restricted to women who were having their first child, and they were between 5 and 40 weeks gestation at the time they completed the instrument. Demographic data were also gathered. RESULTS: (1) Maternal-fatal attachment increased significantly from 5 to 40 weeks of gestation. Especially feeling fetal movement had positive effect on maternal fetal attachment. (2) Women who reported negative perception or ambivalent feeling about their pregnancy showed low attachment score. And women whose husband reported negative feeling about their pregnancy responded lower in the scale. (3) Some negative relationships were observed between maternal-fetal attachment score and the histories of abortion and sterility. (4) Maternal-fetal attachment showed no significant correlations to factors of threatened abortion, premature labor, and IUGR. (5) Maternal-fetal attachment showed negative correlations to State-Trait anxiety during early pregnancies. PMID- 8242154 TI - [A study of falls among elderly living in the community]. AB - This study was conducted to mainly investigate the causes of falls among elderly living in the community and also to clarify the status of the elderly one year after they experienced falls. Subjects who had lived in Koga city, Ibaraki prefecture were chosen. An average age of the subjects (n = 132) was 73.7 +/- 6.4 years. Self administrated questionnaire was employed. The main results were obtained as follows. 1) The average of the falls of a subject for the past one year was 1.75 +/- 2.40 times. About 70 percent of the falls occured outdoor. About 70 percent of the causes of the falls were extrinsic. 2) The higher injuries frequency occured among those who got medical treatment. Diuretic and hypotensive drugs occupide the major two kinds of all. 3) The physical symptoms such as pains, fatigues, walking problems, as well as less active behavior characterized those who experienced the fall-caused external injuries. The falls triggered off deteriorating the elderly's healthy physical status. It was suggested that nursing intervention was very important to prevent falls among elderly in the community to maintain their quality of life (Q.O.L). PMID- 8242155 TI - [Hemodynamic changes in patients with myocardial infarction while walking up and down stairs]. AB - Hemodynamic changes while walking up and down stairs were studied in 53 myocardial infarction (MI) patients in the chronic phase and 31 adults of normal health. Electrocardiograms were recorded by a holter monitor and analyzed for arrhythmias and ST segment changes. Heart rate, systolic and dyastolic pressure, and pressure-rate product (PRP) were also measured by auto sphygmomanometer. These were measured at one minute intervals during the following period: resting in bed for 3 minutes in a supine position; walking to a stairway (1 minute); walking up and down 2 flights of stairs taking 30 seconds for each flight (total of 2 minutes) and returning to their bed (1 minute). All subjects then rested until they reached their starting PRP level. After walking up and down the stairs, the PRP increased significantly over that measured while the subjects were in a supine position for both the MI patients and those of normal health. However, the rate of increase for the MI patients was lower than that of the normal adults. Also, for MI patients older than 60, increases of the PRP were lower than the increases of the younger patients. This phenomenon may be due to low cardiac function, low cardiac reserve, vascular sclerosis, and the hemodynamical instability of the weak vasomoter reflex. In addition, the average recovery time after walking up and down the stairs was 1.9 minutes for MI patients, compared to 1.2 minutes for normal adults. 17% of the patients spent more than 3 minutes recovering, while the recovery times for all normal adults were within 2 minutes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242156 TI - [Factors effecting medication errors]. AB - The purpose of this paper is to reveal factors that may occur or that may prevent medication errors, and to consider some measures of patients' safety. For this aim, we explored the characteristics of each ward and the characteristics of each nurse as well as their experiences about medication errors. The investigation was conducted at a approximately 650-bed university hospital located in a large urban center in Tokyo, and at a approximately 150-bed private hospital located in a suburb of Tokyo. Questionnaires were distributed in October 1991 and asked nurses to identify medication errors in which they had been personally committed within the past one-month. A response rate was approximately 70%. We were informed other information of each unit from head nurses--such as the unit size, case mix, nurse staffing and working schedule of nurses. Results indicate that on the questionnaires the nurses reported 10 times medication errors than they reported to head nurses. 56 percent of 155 nurses (except nurses who were working at an ICU and a mental ward) stated they had made medication errors or near misses during the 1-month reporting period. 30 percent of 155 stated they actually dealt with medication errors occurred by others. Several factors that effect medication errors have been identified by a stepwise discriminant analysis. The proportion of serious patient and the bed occupancy rate influenced the occurrence of medication errors. The educational career of a nurse, especially whether the nurse had been offered an assistant nursing program or not, related to dealing with and/or preventing medication errors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242157 TI - [The study of senile symptoms and home care on the elderly living in Korea]. AB - The purpose of this study is to investigate the daily life and care condition of elderly people living in Korea, and to observe the symptom of senile dementia among them. In the first survey, 713 subjects were screened and 42 subjects were diagnosed as suffering from senile dementia. In the second survey, 26 elderly and their caregivers who agreed to participate in this program, were interviewed. The results were as follows. 1. About 25% of the 713 subjects needed some kind of help in their daily life. 2. 85% of the subjects could go out and about their houses without help. Only 5% of them had a complete or a partial loss of activity. 3. Of all the subjects, 42 were diagnosed as suffering from senile dementia. The prevalence of this disease was calculated at 5.6% in the survey. 4. Each demented elderly person had 5.3 mental symptoms on the average. Depression was observed more among women and violence was observed more among men. 5. The elderly who had a lower CPR score (Caretaker-Patient Relationship Score), indicating a poor relationship between the patient and caretaker, had a higher prevalence of mental symptoms than those who had a higher CPR score. PMID- 8242158 TI - Is there a role for diet or other questionable therapies in managing rheumatic diseases? PMID- 8242159 TI - Methotrexate in rheumatoid arthritis. PMID- 8242160 TI - Pathogenesis of osteoporosis. PMID- 8242161 TI - Prevention of osteoporosis. PMID- 8242162 TI - Exercise strategies for osteoporosis. PMID- 8242163 TI - Bond strength of composite to alloy treated with bonding systems. AB - PURPOSE: The in-vitro bond strengths of a composite bonded to a nickel-chromium beryllium alloy treated by eight bonding systems were measured after three storage conditions. MATERIALS AND METHODS: Nickel-chromium-beryllium alloy samples were treated by eight commercial bonding systems including adhesive composite cements, all-purpose bonding agents, and silica-coating systems. A composite was bonded to the alloy samples. Sample groups were stored in water for 24 hours at 23 degrees C, or thermocycled, or stored for 6 months at 23 degrees C and then debonded in tension. RESULTS: Bond strengths after 24 hours storage were: adhesive composite cements, 14.2 to 22.1 MPa; all-purpose bonding agents, 11.4 to 14.6 MPa; and silica-coating systems, 18.6 to 20.2 MPa. Bond strengths after thermocycling were: adhesive composite cements, 12.6 to 20.6 MPa; all purpose bonding agents, 9.9 to 17.7 MPa; and silica-coating systems, 11.1 to 19.2 MPa. Bond strengths after 6 months were: adhesive composite cements, 12.0 to 13.1 MPa; all-purpose bonding agents, 8.7 to 14.1 MPa; and silica-coating systems, 14.8 to 18.4 MPa. CONCLUSIONS: Only two bonding systems showed decreased bond strength after thermocycling, as compared with 24 hours' storage; whereas four bond systems showed decreased bond strength after 6 months' storage. Bond strengths measured after thermocycling were not predictive of those measured after 6 months' storage. PMID- 8242164 TI - The effect of fatigue on the shear bond strength of resin bonded to porcelain. AB - PURPOSE: The objective of this study was to determine whether fatigue cycling affects the shear bond strength of a resin that is initially strongly bonded to porcelain. MATERIALS AND METHODS: Thirty-five disks of a feldspathic/leucite porcelain were fired on a phosphate-bonded investment material. Each disk was etched with hydrofluoric acid and treated with a silane solution and an adhesive. Resin cement composite cylinders were applied to the treated porcelain and light cured. All specimens were stored for 1 week in distilled water at 37 degrees C and then thermocycled for 1000 cycles in water between 2 degrees C and 50 degrees C. Fifteen of the specimens were randomly selected as control specimens. The remaining were subjected to fatigue cycling. A shear load was applied parallel to the bonded surface and cycled in a square wave between 0 and 26 N at 2 Hz for 27,500 cycles. All specimens were fractured in shear at 1.27 mm/min, and the shear bond strength was determined. RESULTS: Fifteen percent of fatigued specimens and 6.7% of the control specimens showed at least partial adhesive failure. All other specimens in both groups failed cohesively in the porcelain. There was no significant difference in the mean bond strengths of the fatigued and control groups (analysis of covariance, P > .05). CONCLUSIONS: For the conditions investigated, cyclic fatigue did not reduce the bond strength of this resin/porcelain system. PMID- 8242165 TI - Osteoporosis and osseointegration of implants. AB - According to medical literature, osteoporosis and related bone pathologies are increasing in epidemic proportions. The exact etiology of the disease is unknown, but hormonal, dietary, and genetic factors all contribute to the related loss of bone density. In the disease process, bone loss occurs throughout the body. Research indicates that the mandible and maxilla are affected, and show oral manifestations. There is no scientific data to contraindicate the use of two-step osseointegrated implants in osteoporotic individuals. The purpose of this article is to review the literature regarding osteoporosis and its relationship to oral bone loss. PMID- 8242166 TI - Clasp retention: the effects of five variables. AB - An attempt was made to determine the relative retention of each of the four most commonly used clasp assemblies. They are the rest, proximal plate, I bar (RPI); rest, proximal plate, facial circumferential arm (RPA); modified T; and Akers (facial and lingual circumferential arms). A mandibular bilateral distal extension base removable partial denture design was selected because it is most commonly used. A test model was made by removing all mandibular molars from a Columbia Dentoform and replacing the ivorine second bicuspids with natural teeth. These, in turn, were later replaced with gold crowns that were identical in contour to the natural teeth. Eight removable partial dentures were constructed using preformed plastic lingual bars, minor connectors, denture base retention loops, and clasps. Artificial teeth and the denture bases were processed to each framework as for a clinical patient. Each removable partial denture was placed in the Instron testing machine and pulled from the test model. The investigation examined the effectiveness of each of four types of clasps in resisting displacing forces, in both vertical (occlusal) and mesio-occlusal directions. Each clasp was tested using natural teeth and gold crowns, in dry and wet environments, both with and without indirect retainers. Results indicate that, (1) The RPA clasp was the most retentive of all on natural abutments, against both directions of pull, but only slightly so over the RPI clasp with a vertical pull. (2) The Akers clasp was most retentive against a mesio-occlusal pull on the gold abutments. (3) Natural tooth structure gives slightly more frictional resistance than a gold surface.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242167 TI - Alternative barrier technique. AB - This article offers an alternative barrier technique that not only places the barrier in the dental treatment room, but also places it under the direct supervision and control of the dentist. A known hepatitis B carrier was selected for immediate denture treatment to illustrate and test this proposed barrier technique. PMID- 8242168 TI - Oral electrical burns: etiology, histopathology, and prosthodontic treatment. AB - Oral electrical burns occur predominantly in young children and may lead to permanent disfigurement. The etiology and histopathology of oral electrical burns are discussed. Prosthodontic treatment options are presented to provide clinical guidelines for the practitioner. PMID- 8242169 TI - Restorative and prosthetic dental education: changes, challenges, and opportunities. PMID- 8242170 TI - The accuracy of six impression techniques for osseointegrated implants. AB - PURPOSE: Six impression techniques were evaluated using tapered and square impression copings. MATERIALS AND METHODS: The absolute distortion was measured using a cast metal impression tray with fiduciary reference points external to the impression material. Measurements of the x, y, z coordinates on the master cast and the impressions were directly made with a travelling digitizing microscope. The difference between the coordinates of each of six sites in the impression and the corresponding reference site were compared. RESULTS: There was no significant difference between the techniques for the square copings but that there was a significant loss of accuracy in the z-axis with the tapered copings. CONCLUSIONS: No significant differences were noted. PMID- 8242171 TI - Finite element stress analysis of tooth-to-implant fixed partial denture designs. AB - PURPOSE: The purpose of this study was to conduct a three-dimensional finite element stress analysis to compare models representing a natural tooth and an integrated implant connected with rigid and nonrigid prostheses. MATERIALS AND METHODS: The mathematical models described an integrated implant connected to a second premolar tooth with a three-unit metal-ceramic fixed partial denture. In one model, the tooth and implant were rigidly connected, and in the other, a nonrigid connection was assumed. From a simulated vertical load the computer generated resultant stress contours and values (compressive and tensile) in horizontal and vertical dimensions. RESULTS: A comparison of the magnitude and pattern of stresses generated in the models shows minimal differences. Although the maximum stress values were slightly higher in some areas of the rigid prosthesis model, the differences were negligible. CONCLUSIONS: Based on the similarities in both the patterns of stress contours and the stress values generated in the two models, advocating a nonrigid connection because of a biomechanical advantage may be erroneous. PMID- 8242172 TI - Evaluation of strain at the terminal abutment site of a fixed mandibular implant prosthesis during cantilever loading. AB - PURPOSE: Cantilever lengths from 10 mm to 20 mm have been empirically recommended for Branemark fixed mandibular implant prostheses. However, functional stresses generated within the framework and at the crestal bone associated with various cantilever lengths have not been well researched. The purpose of this investigation was to evaluate the strain generated within an implant-supported prosthesis and on a simulated bone surface during functional cantilever loading. MATERIALS AND METHODS: A symmetrical mandibular fixed-implant framework supported by six Nobelpharma 7.0 x 4.0-mm abutments and 15.0 x 4.0-mm fixtures was fabricated. The fixtures were embedded in a simulated bone matrix of polymethyl methacrylate resin. Fourteen different arrangements of active supporting abutments were tested during 15 lb unilateral static cantilever loading 7 mm, 14 mm and 20 mm distal to the terminal abutments. T-rosette strain gauges were placed immediately distal to the terminal abutment site on the right side of the framework and on the corresponding simulated bone surface. RESULTS: There was no difference in framework microstrain as abutment number and arrangement were varied. Microstrain distal to the terminal abutment increased significantly with increasing cantilever length. Distal abutment microstrain increased 213% (63 mu epsilon to 197 mu epsilon) when cantilever length was increased from 7 mm to 14 mm and an additional 55% (197 mu epsilon to 306 mu epsilon) when cantilever length was increased from 14 mm to 20 mm. Overall, microstrain increased 306% when cantilever length was increased from 7 mm to 20 mm. Microstrain on the framework was always tensile (positive). Microstrain at the simulated bone reached higher maximum levels than on the framework (-588 mu epsilon versus 314 mu epsilon) and was compressive in nature (negative). In contrast to framework microstrain, microstrain at the simulated bone site varied dramatically with changes in abutment arrangement. Strains observed at the simulated bone surface increased dramatically as the distance to the adjacent active abutment increased or as the anterior-posterior span of abutments decreased. Distal abutment microstrain also increased significantly at the bone site as cantilever length increased, however, percent increases were less (7 mm to 14 mm, 55%; 14 mm to 20 mm, 30%; 7 mm to 20 mm, 101%). CONCLUSIONS: The results of this study indicate that an optimum biomechanical environment should exist when cantilever spans exceeding 7 mm are planned regardless of the number of supporting abutments. Strain transmitted to the crestal bone can be decreased by maximizing the number and anterior-posterior spread of supporting fixtures while minimizing the distance between the distal abutment and its adjacent abutment. PMID- 8242173 TI - Desmin pathology in neuromuscular diseases. AB - Desmin is an intermediate filament protein that in striated muscle is normally located at Z-bands, beneath the sarcolemma, and prominently at neuromuscular junctions. It is abundant during myogenesis and in regenerating fibers, but decreases in amount with maturation; in regenerating and denervated muscle fibers it is co-expressed with vimentin. Aggregates of desmin occur as nonspecific cytoplasmic bodies or cytoplasmic spheroid complexes, similar to the aggregates of keratin filaments in Mallory bodies or the neurofilament aggregates in Lewy bodies. In all three instances, alpha-B crystallin may be associated with desmin. There are now increasing numbers of neuromuscular disorders in which abnormal amounts of desmin, some abnormally phosphorylated, feature prominently in muscle fibres. Several of these diseases, including spheroid body myopathy, granulo filamentous body myopathy and the dystrophinopathies, are familial. Ultrastructural and immunohistochemical studies of desmin have considerably broadened our understanding of the pathology of the cytoskeleton in muscle fibers and in certain hereditary neuromuscular diseases. PMID- 8242174 TI - Cytogenetic differences between intestinal and diffuse types of human gastric carcinoma. AB - This study concerns the cytogenetics of 23 gastric carcinomas, classified histologically as intestinal or diffuse types. In carcinomas of the diffuse type, the only numerical changes observed were Y chromosome loss associated with X chromosome disomy in four of seven male patients. A 46, XX karyotype without recognizable alterations was observed in three of five female patients, and rare structural changes in diffuse carcinomas involved chromosomes 1 and 18. In contrast, intestinal type tumors were exclusively aneuploid, with chromosome modes ranging from 48 to 84. The most consistent change was trisomy 20 in seven of 11 patients, each of which displayed a number of both single and clonal structural aberrations. Frequent structural changes were translocations involving chromosome 13 (including a putative isochromosome 13q in three of 11 patients), and alterations in chromosomes 1, 6 and 12. This study therefore suggests that diffuse and intestinal types of gastric carcinomas do not share a common sequence of genetic changes. The tumor with the worse prognosis (diffuse type) is surprisingly diploid, with uniform X-disomy in both males and females. The clinically less aggressive tumors (intestinal type) show multiple changes, both numerical and structural, of which some are reminiscent of changes seen in tumors of the lower gastrointestinal tract. Cytogenetics may thus be a valuable adjunct in establishing the diagnosis, classification, and prognosis of gastric carcinomas. PMID- 8242175 TI - Virus-neuron interactions in the mouse brain infected with Japanese encephalitis virus. AB - The virus-host interactions between Japanese encephalitis (JE) virus and mouse brain neurons were analyzed by electron microscopy. JE virus replicated exclusively in the rough endoplasmic reticulum (RER) of neurons. In the early phase of infection, the perikaryon of infected neurons had relatively normal looking lamellar RER whose cisternae showed focal dilations containing progeny virions and characteristic endoplasmic reticulum (ER) vesicles. The reticular RER, consisted of rows of ribosomes surrounding irregular-shaped, membrane unbounded cisternae and resembled that observed in JE-virus-infected PC12 cells, were also seen adjacent to the lamellar RER. The appearance of the reticular RER indicated that RER morphogenesis occurred in infected neurons in association with the viral replication. The fine network of Golgi apparatus was extensively obliterated by fragmentation and dissolution of the Golgi membranes and their replacement by the electron-lucent material. As the infection progressed, the lamellar RER was increasingly replaced by the hypertrophic RER which had diffusely dilated cisternae containing multiple progeny virions and ER vesicles. The Golgi apparatus, at this stage, was seen as coarse, localized Golgi complexes near the hypertrophic RER. In the later phase of infection, RER of infected neurons showed a degenerative change, with the cystically dilated cisternae being filled with ER vesicles and virions. Small, localized Golgi complexes frequently showed vesiculation, vacuolation, and dispersion. The present study, therefore, indicated that during the viral replication the normal lamellar RER which synthesized neuronal secretory and membrane proteins was replaced by the hypertrophic RER which synthesized the viral proteins. The hypertrophic RER eventually degenerated into cystic RER whose cisternae were filled with viral products. The constant degenerative change which occurred in the Golgi apparatus during the viral replication suggested that some of the viral proteins transported from RER to the Golgi apparatus were harmful to the Golgi apparatus and that increasing damage to the Golgi apparatus during the viral replication played the principal role in the pathogenesis of JE-virus-infected neurons in the central nervous system. PMID- 8242176 TI - Apoptosis and cell desquamation in repair process of ischemic tubular necrosis. AB - To elucidate the role of apoptosis and cell desquamation in the repair phase of acute tubular necrosis, morphological findings after 60 min ischaemia were investigated in rats. A morphometric analysis of the cell proliferation and of the epithelial cellularity of reconstructing tubules was performed. The kinetics of apoptosis and cell desquamation were also examined. Ischaemia and reperfusion injury resulted in widespread necrosis of tubules at day 1. Subsequently, a regenerative epithelial hyperplasia took place in the early stage. The most marked increase in cellularity in the damaged tubules was on day 6, when the tubules became lined by hyperplastic epithelial cells with papillary clusters. The number of papillary clusters decrease up to day 8, and during this period many desquamated cells from the clusters were observed in the tubular lumen. In the later stage, hyperplastic epithelial cells were reduced to their original cellularity and during this period the number of apoptotic cells obviously increased, while the damaged tubules were reconstructed. We conclude that epithelial overproduction occurs in the early phase after tubular necrosis, and excess hyperplastic epithelial cells regress during the repair process by cell desquamation and apoptosis, both of which are essential for the recovery of the original tubular structure. PMID- 8242177 TI - Clara cell antigen in normal and migratory dysplastic Clara cells, and bronchioloalveolar carcinoma of Syrian hamsters induced by N-nitrosomethyl-n heptylamine. AB - Histogenetic features of lung tumours were studied in Syrian hamsters that had been induced with 6.8 mg N-nitrosomethyl-n-heptylamine/animal by gavage once a week for 35 weeks. At intervals from experimental week 2 until week 46, pulmonary tissues from hamsters were examined by light and electron microscopy. This report describes early hyperplastic lesions associated with terminal bronchioles and the progression of these lesions to bronchioloalveolar tumours. Using immunohistochemical and ultrastructural colloidal gold labelling techniques, hamster Clara cell antigen was found to be localized in Clara cell granules and smooth endoplasmic reticulum of normal cells, in dysplastic Clara cells migrating through basement membrane defects or from the open end of terminal bronchioles, and in hyperplastic peribronchiolar cell foci. The latter progressed to bronchioloalveolar tumours growing out along alveolar basement membranes in a characteristic lace-like, lepidic pattern. Tumours were composed of secretory (Clara), ciliated, mucous, and undifferentiated cells, as well as trapped, non neoplastic alveolar type II cells. Hyperplastic neuroendocrine cell foci lining airways were immunoreactive for chromogranin, but these cells did not participate in the pre-neoplastic or neoplastic process. It is suggested that bronchioloalveolar carcinomas in hamsters are derived from bronchiolar secretory (Clara) cells growing along alveolar walls, differentiating into other bronchiolar cell types and entrapping resident alveolar type II cells. Due to the migratory capacity of Clara cells, it is also possible for tumours composed of bronchiolar cells to develop at the lung periphery. PMID- 8242178 TI - Function and "homing" of the lung macrophages. I. Evidence of functional heterogeneity of mobile cells of the murine lung parenchyma in the bronchoalveolar lavage (BAL). AB - The object of the present investigation was to extend fundamental understanding of the composition, function and origin of bronchoalveolar lavage (BAL) cells, and to reduce the considerable gaps in our existing knowledge. BAL cells from the B10.D2 mouse were identified by morphological, enzyme cytochemical and immunocytochemical methods and a further functional characterisation of macrophages was undertaken in vitro using latex phagocytosis. The possible bone marrow origin of BAL cells was investigated with a radiation chimera. The bone marrow from H2k-positive F1 mice (B10.BR x B10.D2) was transplanted into the irradiated H2k-negative parent animals. At various time intervals after transplantation, BAL cells were tested immunocytochemically for the presence of H2k-positive cells. The principal finding was that BAL cells are functionally heterogeneous. It was possible by morphological, enzyme cytochemical and immunocytochemical techniques to subdivide the total BAL cell count into 89% macrophages, 8.5% lymphocytes and 2.5% granulocytes. Only 14.3% of the macrophages expressed the corresponding epitope, and this small population could be further subdivided immunocytochemically into mature and immature (7:1) macrophages. The ability to phagocytose, which is typical of active macrophages, could be induced in vitro in only one-third of all BAL macrophages. Three quarters of the macrophages carried the H2d antigen and 9.5% of the BAL cells were natural killer cells showing a macrophage phenotype. Subdivision of the lymphocytes showed a significant majority of T-cells (75%) to B-cells (20%), and 5% Asialo GM1-positive cells. The CD4/CD8 ratio amounted to 1.3:1 and 38% of the lymphocytes expressed on the surface the H2d antigen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242179 TI - [The Revista Latino-Americana de Enfermagem]. PMID- 8242180 TI - [The need for nursing research]. AB - The author locates the trajectory of the knowledge in nursing, based on light of professionalism and presents a synthesis of the community effort of nursing in conquering the conditions for the administration of knowledge. The analyze of the beginning of the seventy decade, point out the knowledge explosion in nursing literature, and an advance in the interchange, getting together, so for, the challenge of the over flow from the descriptive phase and the difficulty of research in transforming the practice. PMID- 8242181 TI - [The development of nursing in Latin America: a strategic view]. AB - Nursing in Latin America is facing a number of serious challenges significantly affecting the contributions its personnel con make to the health of the people they serve. This article presents an overview of nursing as an important component of the health system in Latin America through the prospective and orientation provided by strategic planning. The author, following an analysis of the nursing situation in the Region, identifies several problems and then creates a vision for the future. The article looks at the possibilities of developing and implementing projects and action plans to address the priority problems. It sees strategic thinking as a possible means for nursing to reflect on and maximize its contribution and presence in the solution of the health problems in the current decade and into the next century. PMID- 8242182 TI - [The W. K. Kellogg Foundation and nursing development in Latin America]. PMID- 8242183 TI - [Postgraduate nursing education in Brazil--the current situation]. AB - This article describes the actual situation of post-graduate programs in nursing in Brazil based on data for the biennium 90/91, from the Coordination of Post Graduate Programs (CAPES) regarding: the faculty, curricular structure, organization of research, scientific production by the faculty, scientific production by the students, mean time for receipt of degree, and student flux. A global evaluation of post-graduate programs in nursing throughout their 20 years of existence is presented. PMID- 8242184 TI - [Nursing research in Brazil and its position in the Federal Agency for Development]. AB - This study presents a historical retrospective of research activities in nursing in Brazil, with emphasis on systematization, tendencies and diffusion of these studies, especially for the support of these by the National Council of Scientific and Technological Development. The origins and actual structure of this agency is also presented. In discussing the insertion of nursing in this agency of financial support, the contradictory situation between the official proposal for the training of a nurse and the orientation offered by the National Council of Scientific and Technological Development is reported. This study also offers reflection about the importance of the definition of research lines for nursing, in view of the actual demands of the process of knowledge production. PMID- 8242185 TI - [Nursing: the reality and future in care and management]. AB - The authors discuss the activities developed by Nursing in two general orientations: the assistance and the management, concerning the concrete work and its perspectives. Therefore, they engage in polemics around the administrative versus assistance functions and attract the attention to the impossibility of thinking the nursing work inarticulated of different health practices, as a whole they accomplish each other in attempting the necessities of health of the population in a social level. PMID- 8242186 TI - [We live in a society of information]. PMID- 8242187 TI - [An alternative methodology for the teaching of anatomy in nursing studies at the University of Carabobo]. AB - An alternative methodology for the teaching of Human Anatomy which contributes to improve academic yield by Nursing students at the Faculty of Health Sciences of the University of Carabobo is presented. A total of 50 first-year Nursing students who participated in the experiment were divided into 2 groups, control and experimental. This research project was of the almost-experimental type and the work methodology was experimental-evaluation. Three instruments were used to obtain data: an investigation of the Lickert school type consisting of 40 proposals related to the students attitude concerning the study of this discipline and in view of a teaching methodology (EAEAM), accompanied by the school of Interests of Thurstone and by the Psychometric Test of Raven, to determine relationships in accordance with the research objectives. In function of the empirical results of the experiment and of revision of the literature pertaining to this theme, a methodological proposal is presented for the teaching of Human Anatomy; its theoretical-practical nature demands: previous study, observation, discussion and, fundamentally, subdivision, as an alternative to improve academic yield by Nursing students. PMID- 8242188 TI - [The postgraduate program at the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242189 TI - [The Department of Psychiatric Nursing and Human Sciences of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242191 TI - [The Commission on Culture and University Extension of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242190 TI - [The Department of General and Specialized Nursing of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242192 TI - [The Commission on Research of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242193 TI - [The Commission on Graduation of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242194 TI - [On 18 December 1992 at the Ribeirao Preto School of Nursing of the University of Sao Paulo the Revista Latino-Americana de Enfermagem was launched]. PMID- 8242195 TI - [The nursing care of children presenting with Down's syndrome]. AB - Diagnosis of Down Syndrome can be made in the nursery. The presence of an atypical child often leads to disaggregation of the family. Nursing intervention teaching early stimulation, hygiene and feeding techniques permits better adjustment. The study of 10 Down Syndrome children and their families during o period of 6 years, made it possible to show that immediate diagnosis of the defect leading to early stimulation of the child and counselling of the couple led to more favourable results. Later stimulation was found to cause a more difficult adaptation of the child within the family. PMID- 8242196 TI - [The Spanish influence in Colombian nursing]. AB - Four statements inferenced from: religious traditions, gender or woman status, military heritage and ethnic inheritance, and their influence in nursing, are presented in this article. The ethnic inheritance analyses the issue based upon the cultural influences of the native-indigenous groups and the spanish and mediterranean attributes in nursing development. The religious tradition began with Pedro Claver's J.C. presence between 1610 and 1617. His presence is followed by the "Hermanos Hospitalarios de San Juan de Dios" in 1768, and further with the presence of the sisters of Charity in 1873. Lastly, the article compares the Barcelona's Santa Cruz hospital organization, where at that time, seems to appear a new type of nursing arrangement; with present functions and charges, currently utilized in colombian hospitals (administrator of patient rooms, "servidor", women in charge of female patients and sick children, "economa", chief in charge, and senior male nurse). PMID- 8242197 TI - [The Department of Psychiatric Nursing and Human Sciences of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242198 TI - [The Department of Maternal-Child Nursing and Public Health of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242199 TI - [The Commission on Postgraduate Studies of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242200 TI - [The Commission on Culture and University Extension of the Ribeirao Preto School of Nursing of the University of Sao Paulo]. PMID- 8242201 TI - [The work of the nurse in a surgical center--an analysis of the data collected]. PMID- 8242202 TI - [Curricular reform of undergraduate nursing--the Ribeirao Preto School of Nursing, the University of Sao Paulo. The Commission on Curriculum Restructuring of the Ribeirao Preto/USP School of Nursing]. AB - The objective of the present study was to describe the process of curricular renovation for the undergraduate Nursing course developed by the Nursing School of Ribeirao Preto, USP, with emphasis on the following planning stages: diagnosis, educational objectives, selection and organization of content, curricular organization, curricular strategies, and curricular evaluation. In view of a diagnosis of current reality based on a philosophical-pedagogic approach which demonstrates the need to train general duty nurses, we present a conceptual framework directed at the view of man, community, health-sickness process, nursing, general duty nurses, competence, and curriculum. We propose a stronger emphasis on the teaching of biological and human sciences in the basic cycle, as well as on administrative aspects in sanitary and hospital activities. This curriculum covers the cycles of preprofessional and professional training to be offered over eight semesters in the undergraduate Nursing course at EERP-USP. PMID- 8242203 TI - [40 years old. Maturity conquered]. AB - In this narrative, I do not plan to be a cold spectator who captures and relates facts for future use, but one who "lived" each moment. This is also a contribution to history, once one tries to illuminate the present remembering the happenings of 4 decades (1953 to 1993) of the College of Nursing at Ribeirao Preto, University of Sao Paulo, focusing on the beginning, the changes, the adjustments and perspectives. PMID- 8242204 TI - [The access, utilization and acceptance of dermatology services at a teaching health center from the viewpoint of Hansen's disease patients]. AB - The present Case Study intends to analyse the access, utilization and acceptance of the Dermatology services of a SHC based mainly in the orally reported histories of 27 patients affected by Hansen's disease. The access to SHC- concerning its organization--took place without difficulty, except for the attendance services. However, the utilization level of the services was found to be higher than the established pattern of the care provided whether in actual episodes of the disease or not. The acceptance of the services rendered was consensual and both the technical quality and interpersonal relationship were emphasized. PMID- 8242205 TI - [The teaching of patient evaluation: the delineation of the content of nursing diagnosis]. AB - The nursing care is based on the nursing assessment. In the nurse's professional activity there aren't clarity and consensus about the clinical entities, problems or phenomenons that nurses intended to care. Difficulties in deciding what should be collected are determined by this indefinition, that it is also present in nursing graduate courses. In this article the authors discuss that the Nursing Diagnosis can be the framework for the nursing assessment. PMID- 8242206 TI - [Pediatric nursing knowledge: books edited in Brazil from 1916 to 1988]. AB - From the apprehension of the knowledge in pediatrics nursing in Brazil, a research was done in a Brazilian historical bibliographic work of references. From an historical perspective nursing it is seen as a social articulate practice, as part of a collective process of work. The knowledge in this study is considered as an instrument from which man apprehends his working object. Forty titles were found about how to take care of children, from eight per cent of the a group of five hundred and four. The proportion was considered very small. The production which was found was classified under two aspects: the prescriptive and the analytical. The prescriptive texts are those related to normalize the work in view of the rationality and to the productivity. The analytical opens the possibility and give the means to diversify the nursing assistance particularizing the care and with another aim related to the nursing work. PMID- 8242207 TI - [Ecological awareness and health services wastes]. AB - Environmental clean-up involves a series of factors, among them the question of solid wastes, which are not always properly managed. The problem is more serious with respect to medical waste. The awareness of people and the attention of administrators are of fundamental importance for an equilibrated coexistence with the physical environment. PMID- 8242208 TI - The reliability and validity of the COPDSC: a tool that measures self-care behavior of persons with chronic obstructive pulmonary disease. PMID- 8242209 TI - A review of 'fluctuation in mixed venous oxygen saturation in critically ill medical patients: a pilot study. PMID- 8242210 TI - What kinds of strategies exist for staff development in a world of shrinking budgets? PMID- 8242211 TI - Perinatal bereavement counselling: how it has changed. PMID- 8242212 TI - From alternative to complementary: changing the view on the fringe. PMID- 8242213 TI - Herpesvirus infections in childhood: 2. AB - Infections due to herpesviruses have received increasing attention over the past decade, culminating in the isolation in 1986 of human herpesvirus-6. This is the second of two articles in which we examine the clinical spectrum of acquired herpesvirus infections in children and review developments in our understanding of the molecular biology, pathogenesis, treatment and prevention of these infections. PMID- 8242214 TI - Localized cerebral atrophy: an important cause of non-Alzheimer's dementia. AB - Localized forms of cerebral atrophy (lobar atrophies) are more common than textbooks suggest and are often misdiagnosed. They may give rise to conductor disorder and personality change, or progressive language dysfunction. This article outlines the presenting features, the differential diagnosis and appropriate investigations. PMID- 8242215 TI - Immunological tests in autoimmune rheumatic disease. AB - Autoantibodies are the hallmarks of autoimmune diseases. They are not, however, necessarily pathognomonic and interpretation of autoantibody results must be considered in the light of the clinical history. This article highlights the clinically relevant autoantibodies associated with autoimmune rheumatic diseases, and introduces the concept of autoantibody disease imprinting. PMID- 8242216 TI - Spontaneous rupture of the Achilles tendon: pathology and management. AB - Rupture of the Achilles tendon, one of the strongest tendons in the body, can cause significant long-term morbidity if diagnosis or treatment is delayed. This article discusses the current debate concerning the ideal management of these injuries, as well as their pathology and diagnosis. PMID- 8242217 TI - Counselling: indications and results. AB - There are a number of distinct theoretical approaches to counselling. The most influential are behavioural, cognitive-behavioural, psychodynamic and person centered treatments. However, the factors responsible for the beneficial effects of treatment remain to be discovered. PMID- 8242218 TI - History taking. AB - History taking is the most important component of the consultation. It requires the mastery of a defined body of knowledge, skills and attitudes, which do not improve simply with experience and need to be updated. PMID- 8242219 TI - Opioids may cause not relieve the problems. PMID- 8242220 TI - Iatrogenic osteomyelitis following closed cardiopulmonary resuscitation. PMID- 8242221 TI - Fatal pulmonary embolus in a keep-fit enthusiast. PMID- 8242222 TI - Operative viva A. PMID- 8242223 TI - Weaning from mechanical ventilation. PMID- 8242224 TI - Brain biopsy in a patient with bullous lung disease. PMID- 8242226 TI - PAF activation of a voltage-gated R-type Ca2+ channel in human and canine aortic endothelial cells. AB - By the use of fura-2 and digital imaging techniques, [K]o depolarization or PAF (10(-9) M) were shown to induce a sustained increase of [Ca]i in human or canine single aortic vascular endothelial cells (VEC) that was insensitive to nifedipine but sensitive to (-)-PN200-110 or to lowering of [Ca]o. The PAF-induced effect on [Ca]i was blocked by the PAF receptor antagonist, WEB2170. Our results suggest that [K]o depolarization and PAF increase [Ca]i via the activation of R-type Ca2+ channels. PMID- 8242225 TI - Fourteenth Gaddum Memorial Lecture. A current view of tamoxifen for the treatment and prevention of breast cancer. AB - Tamoxifen has been found to be a safe and effective treatment for all stages of breast cancer. Long term tamoxifen therapy is associated with some rare, but potentially serious, side effects so patients should be carefully monitored. However, long term tamoxifen therapy is also associated with a number of physiological benefits over and above its tumouristatic action. These benefits include a decrease in the development of contralateral breast cancer, the maintenance of bone density in postmenopausal women and a decrease in cardiovascular disease. The successful application of tamoxifen to treat breast cancer has increased enthusiasm to test its worth to prevent breast cancer. Although there are individual requests by patients for tamoxifen to prevent breast cancer, individual treatment is inappropriate. Tamoxifen can only be adequately evaluated as a preventive in randomized, double-blind clinical trials. These trials are in place and physicians should encourage women to participate and establish a new therapeutic option as rapidly as possible. PMID- 8242227 TI - Differential class III and glibenclamide effects on action potential duration in guinea-pig papillary muscle during normoxia and hypoxia/ischaemia. AB - 1. Microelectrode recording techniques were used to study the effects of several potassium channel blockers which are considered to be Class III antiarrhythmic compounds. The effects of (+)-sotalol, UK-66,914, UK-68,798 and E-4031 on action potential duration (APD) were determined in guinea-pig isolated papillary muscles. The compounds were evaluated under normoxic or hypoxic/ischaemic conditions at 36.5 degrees C and compared to glibenclamide, which is considered to be a blocker of ATP-dependent potassium channels. Prolongation of action potential duration at 90% repolarization (APD90) was taken as an indirect measure of potassium channel blockade. 2. Under normoxic conditions, the Class III compounds prolonged APD in a concentration-dependent manner. According to EC15 values, the order of potency of the Class III compounds was found to be UK-68,798 > E-4031 > UK-66,914 > (+)-sotalol. Glibenclamide did not significantly prolong APD90 under normoxic conditions. 3. Perfusion with an experimental hypoxic or ischaemic bathing solution produced qualitatively similar effects on action potentials. Over a period of 20-25 min in either of the experimental solutions, there was a small decrease in action potential amplitude (APA) and a prominent shortening of APD. The ischaemic solution also depolarized the resting membrane potential by about 15 mV. 4. (+)-Sotalol and UK-66,914 did not reverse the shortening of APD induced by perfusion with hypoxic Krebs solution. High concentrations of glibenclamide (10 microM) and UK-68,798 (30 and 60 microM) partially reversed the hypoxia-shortened APD. Glibenclamide was more potent and exhibited a greater time-dependent action than UK-68,798. 5. During experimental ischaemia, the Class III compound E-4031 (10 microM, n = 7) produced small, but significant, increases in the APD90 (11 +/-3 ms after 20 min) which were not clearly time-dependent(14 +/- 4 ms after 30 min). UK-68,798 (10 microM) also produced a small, but insignificant, increase in APD90(12 =/-6 ms at 20 min, n = 4). Higher concentrations of UK-68,798 (30 and 60 microM, n = 4) did not produce a consistently significant increase in APD90 during ischaemia: significance was only attained after 20 min in the presence of 60 microM UK-68,798 (24 +/- 12 ms). However, in marked contrast to the effects of the Class III compounds, glibenclamide (10 microM) produced large time-dependent increases in ischaemic APD90 (34 +/- 11 ms at 7 min, n = 9) which were significant 15 min or more after drug addition(52 +/- 12 ms at 20 min, n = 7; 74 +/- 5 ms at 30 min, n = 6).6. The present microelectrode data suggest that blockers of ATP-dependent potassium channels, such as glibenclamide, might prove to be more effective than Class III compounds against ischaemia-induced shortening of cardiac action potentials. PMID- 8242228 TI - Characteristics of the binding of [3H]-GR32191 to the thromboxane (TP-) receptor of human platelets. AB - 1. The interaction of the specific thromboxane (TP-) receptor blocking drug, [3H] GR32191 with human intact platelets and platelet membranes has been investigated in vitro. 2. On intact platelets, association of specific [3H]-GR32191 binding at 37 degrees C was biphasic, with an initial rapid component and a slower secondary phase. Dissociation experiments indicated displacement from two sites with t1/2 values of 8.1 and 65.6 minutes. Kd values derived from the kinetic rate constants for the rapid onset/offset and slow onset/offset phases were 0.4 and 0.5 nM respectively. 3. Competition binding of [3H]-GR32191 and GR32191 on intact platelets gave an IC50 of 2.3 nM. Scatchard analysis indicated a single class of binding site with a Kd of 2.2 nM. Further analysis of the data yielded a Hill slope of -1.0 again indicating an interaction at a single binding site. Saturation binding experiments gave a similar estimate of the Kd value for [3H] GR32191 to that obtained from competition binding experiments. A possible explanation for the biphasic interaction of the GR32191 in intact platelets may lie in restriction of its access to and egress from a population of TP-receptors. 4. In platelet membranes at 37 degrees C, specific [3H]-GR32191 binding was complete within 5 min with a calculated association rate constant of 3.2 x 10(8) M-1 min-1. Dissociation of [3H]-GR32191 was relatively slow, with measurable specific binding persisting for > 40 min. Analysis of these data yielded a t1/2 of 17.7 min and a dissociation rate constant of 0.04 min-1 and indicated dissociation from a single site. The ti for dissociation appeared to be related to the contact time of platelet membranes with [3H]-GR32191.Derivation of a Kd from the kinetic rate constants gave a value of 0.13 nM.5. Competition binding of [3H]-GR32191 and GR32191 to platelet membranes gave an IC50 value of 3.5 nM. Scatchard analysis of these data indicated a single binding site with a Kd of 2.1 nM. Saturation binding experiments with [3H]-GR32191 yielded similar IC50 and Kd values to those from competition experiments.6. In further competition binding experiments, the TP-receptor agonists U-46619, STA2, EP171 and 9,1 1-azo PGH2 and antagonists SQ29,548, BM 13.177 and EP092 all competed with specific [3H]-GR32191 binding on intact platelets and, where determined, on platelet membranes. All compounds fully displaced specific [3H]-GR32191 binding. However, where tested, the ICso values for a particular compound were always greater when [3H]-GR32191 was the radioligand than when [3H]-SQ29,548 was used. At the concentrations used in these studies (2 and 5 nM respectively), platelets appeared to bind approximately twice as much [3H]-GR32191 as [3H]-SQ29,548.7. In conclusion, the interaction of [3H]-GR32191 with human intact platelets was complex but the data were consistent with an action at a single class of binding site; from competition experiments this appears to be the functional TP-receptor. The interaction of the drug with this binding site is, however,characterized by a slow dissociation. This characteristic was confirmed in studies with platelet membranes and does not therefore appear to be an artefact of diffusion. Estimates of the Kd of the drug differed depending on the method of determination. Because of the slow dissociation of [3H]-GR32191,those relying upon equilibrium of the radioligand with competing agent may be unreliable. The rate of dissociation also appeared to be related to the contact time of drug with receptor. An explanation for this phenomenon may lie in the ability of GR32191 to induce a change in the conformational state or location of the human platelet TP-receptor. PMID- 8242229 TI - Reduction in the number of thromboxane receptors on human platelets after exposure to GR32191. AB - 1. Exposure of human resuspended platelets in vitro for 30 min to the potent thromboxane A2 (TP)-receptor blocking drug GR32191, followed by its removal by dilution-dissociation, reduced the degree of subsequent binding to 2 nM [3H] GR32191 by almost 50%. Exposure for longer periods (60 min) led to a further reduction. However, no change in the Kd of the radioligand was observed. 2. This effect of GR32191 could not be explained by persistent binding of drug to platelets since a dilution-dissociation stage, designed to remove all drug, was included prior to measurement of binding. 3. Using an alternative TP-receptor radioligand, [3H]-SQ29,548, to monitor receptor number, a reduction in Bmax was observed after GR32191 pre-treatment; the Kd value of the radioligand remained unchanged. 4. The effect was not a common property of TP-receptor blocking drugs since pre-exposure of platelets in vitro for 30 min to BM13.177 or SQ29,548 did not produce a fall in subsequent Bmax to [3H]-SQ29,548. 5. While the mechanism behind this apparent down-regulation of platelet TP-receptor is unknown, it may explain the long duration of action of GR32191 upon platelets in man which persists in the absence of detectable drug in the plasma. PMID- 8242230 TI - Prejunctional modulation of the nitrergic innervation of the canine ileocolonic junction via potassium channels. AB - 1. The effects of different K+ channel blockers were studied on nitric oxide (NO) mediated non-adrenergic non-cholinergic (NANC) relaxations in the canine ileocolonic junction. 2. The non-selective blockers of K+ channels, 4 aminopyridine (4-AP) and tetraethylammonium (TEA) and the blocker of large conductance Ca(2+)-activated K+ channels, charybdotoxin, potently enhanced the NANC relaxations induced by low frequency stimulation. The blocker of small conductance Ca(2+)-activated K+ channels, apamin, had no effect on electrically induced NANC relaxations. 3. NANC nerve-mediated relaxations induced by adenosine 5'-triphosphate (ATP), acetylcholine (ACh) and gamma-aminobutyric acid (GABA) were significantly enhanced by 4-AP and charybdotoxin but not by apamin. TEA significantly enhanced the NANC relaxations in response to GABA and ATP while that in response to ACh was abolished. 4. None of the K+ channel blockers had an effect on the dose-response curve to NO, on the noradrenaline-induced contraction or on the relaxation to nitroglycerine (GTN). 5. From these results we conclude that inhibition of prejunctional K+ channels increases the nitrergic relaxations induced by electrical and chemical receptor stimulation of NANC nerves and thus suggests a regulatory role for these prejunctional K+ channels in the release of NO from NANC nerves in the canine ileocolonic junction. PMID- 8242231 TI - Cyclopiazonic acid, an inhibitor of Ca(2+)-ATPase in sarcoplasmic reticulum, increases excitability in ileal smooth muscle. AB - 1. Effects of cyclopiazonic acid (CPA), a specific inhibitor of Ca(2+)-ATPase in endo- and sarcoplasmic reticulum (ER/SR), on contractile responses, cytosolic Ca2+ concentration and spontaneous electrical activity were examined in ileal longitudinal smooth muscle strips. 2. After intracellular stored Ca2+ in intact ileal strips was depleted by application of 25 mM caffeine in Ca(2+)-free solution, Ca(2+)-loading was performed in the absence or presence of 10 microns CPA in a standard solution containing 2.2 mM Ca2+. Subsequent application of caffeine in Ca(2+)-free solution induced a phasic contraction which was significantly smaller in the strip pretreated with CPA than that in the control. 3. Spontaneous and 20 mM K(+)-induced contractions in the presence of 1 microM atropine were markedly enhanced by 1-30 microM CPA, whereas that induced by 80 mM K+ was not. The magnitude of repetitive transient elevation of cytosolic Ca2+ concentration ([Ca2+])i) and concomitant phasic contractions were markedly enhanced by CPA. The effects were abolished by 10 microM verapamil and restored by 10 microM Bay K 8644. 4. Application of 10 microM CPA depolarized the cell by about 5 mV, decreased the action potential (AP) afterhyperpolarization and markedly increased the frequency of spontaneous AP. These effects were mimicked by 100 nM charybdotoxin. 5. The rate of decay of [Ca2+]i and tension after the bathing solution was changed from one containing 140 mM K+ and 2.2 mM Ca2+ to one containing 5.9 mM K+ and 0 mM Ca2+ was significantly slowed when 10 microM CPA was added to the latter solution. 6. These results indicate that CPA enhances ileal smooth muscle excitability and increases Ca2+-influx through voltage dependent Ca2+ channels. The effect may be consistent with the hypothesis that CPA-induced decrease in stored Ca due to Ca-pump inhibition reduces the Ca2+ dependent K+ current and indirectly enhances Ca2+-influx through membrane activity resulting from the increased excitability.Direct evidence for the regulation of Ca2+ channel activity by intracellular Ca storage sites was not obtained in the present study. PMID- 8242232 TI - K channel activation by nucleotide diphosphates and its inhibition by glibenclamide in vascular smooth muscle cells. AB - 1. Whole-cell and inside-out patch recordings were made from single smooth muscle cells that had been isolated enzymatically and mechanically from the rabbit portal vein. 2. In whole-cells the inclusion in the recording pipette solution of nucleotide diphosphates (NDPs), but not tri- or monophosphates, induced a K current that developed gradually over 5 to 15 min. Intracellular 1 mM guanosine 5'-diphosphate (GDP) induced a slowly developing outward K-current at -37 mV that reached a maximum on average of 72 +/- 4 pA (n = 40). Half maximal effect was estimated to occur with about 0.2 mM GDP. Except for ADP, other NDPs had comparable effects. At 0.1 mM, ADP was equivalent to GDP but at higher concentration ADP was less effective. ADP induced its maximum effect at 1 mM but had almost no effect at 10 mM. 3. In 14% of inside-out patches exposed to 1 mM GDP at the intracellular surface, characteristic K channel activity was observed which showed long (> 1 s) bursts of openings separated by longer closed periods. The current-voltage relationship for the channel was linear in a 60 mM:130 mM K gradient and the unitary conductance was 24 pS. 4. Glibenclamide applied via the extracellular solution was found to be a potent inhibitor of GDP-induced K current (IK(GDP)) in the whole-cell. The Kd was 25 nM and the inhibition was fully reversible on wash-out. 5. IK(GDP) was not evoked if Mg ions were absent from the pipette solution. In contrast the omission of extracellular Mg ions had no effect on outward or inward IK(GDP). 6. Inclusion of 1 mM ATP in the recording pipette solution reduced IK(GDP) and also attenuated its decline during long (25 min) recordings. 7. When perforated-patch whole-cell recording was used, metabolic poisoning with cyanide and 2-deoxy-D-glucose induced a glibenclamide sensitive K-current. This current was not observed when conventional whole-cell recording was used. Possible reasons for this difference are discussed. 8. These K channels appear similar to ATP-sensitive K channels but we refer to them as nucleotide diphosphate-dependent K channels (KNDP) to emphasise what seems to be a primary role for nucleotide diphosphates in their regulation. PMID- 8242233 TI - Single channel and whole-cell K-currents evoked by levcromakalim in smooth muscle cells from the rabbit portal vein. AB - 1. Single channel and whole-cell current recordings were made from single smooth muscle cells isolated from the rabbit portal vein. 2. Application of 10 microM levcromakalim ((-)-Ckm) to single cells held with pipettes containing 1 mM GDP induced a K-current (IK(Ckm)) which occurred in addition to the current caused by GDP alone (IK(GDP)) and averaged 135 pA at -37 mV. We have investigated whether the same K channels underlie the GDP- and Ckm-induced K-currents. 3. If 1 mM GDP was in the pipette but Mg ions were omitted the effect of GDP was absent and IK(Ckm) averaged only 10 pA, suggesting that the action of (-)-Ckm was Mg dependent. 4. Intracellular ATP was not observed to have much effect on IK(-Ckm). Loading of cells with 10 mM ATP from the recording pipette had no significant effect and flash photolysis of caged-ATP loaded into cells from the pipette, estimated to release about 1 mM free ATP, also had no effect on IK(-Ckm). 5. Bath applied glibenclamide inhibited IK(-Ckm) with an IC50 of 200 nM, a value 8 times higher than that found for inhibition of IK(GDP). The delayed rectifier K-current (IK(DR)) was also inhibited by glibenclamide but at higher concentrations (IC50 100 microM). Bath-applied tetraethylammonium ions (TEA) inhibited IK(-Ckm) and IK(GDP) to the same extent (IC50 about 7 mM). 6. In inside-out patch recordings ( )-Ckm (10 microM) applied to the intracellular surface of the membrane potentiated the opening of K channels already stimulated by I mM GDP and all of the channel activity was abolished by 10 microM glibenclamide. The unitary conductance of the channels was 24lpS in a 60 mM: 130 mM K-gradient.7. We suggest that (-)-Ckm may hyperpolarize and relax smooth muscle cells by opening KNDP, a class of small conductance K channels that are related to the ATP-sensitive K channels seen in other tissues. PMID- 8242234 TI - Monocrotaline pyrrole-induced changes in angiotensin-converting enzyme activity of cultured pulmonary artery endothelial cells. AB - 1. Changes in the structural and functional integrity of endothelium have been recognized as relatively early features of delayed and progressive pulmonary vascular injury caused by the pyrrolizidine alkaloid, monocrotaline (MCT). Although a number of investigators have evaluated angiotensin-converting enzyme (ACE) activity in the lungs of rats treated with MCT, the exact nature of changes in activity of this enzyme and the role they may play in MCT pneumotoxicity remain controversial. 2. We examined the direct effects of monocrotaline pyrrole (MCTP), a toxic metabolite of MCT, on cultured endothelial cell ACE activity. Post-confluent monolayers of porcine or bovine pulmonary artery endothelial cells (PECs or BECs, respectively) were treated with a single administration of MCTP at time 0; then they were examined for their ability to degrade the synthetic peptide, [3H]-benzoyl-Phe-Ala-Pro. 3. In PECs, which are relatively insensitive to the direct cytolytic effects of MCTP, monolayer ACE activity was unchanged initially but gradually decreased within 4 days after treatment with a high concentration of MCTP (150 microM). This decrease was transient, and PEC monolayer ACE activity returned to the control value by 10 days post treatment. 4. BEC monolayer ACE activity was also unchanged initially but rapidly declined within 4 days after MCTP treatment and remained depressed throughout the post treatment period. BECs were quite sensitive to the cytolytic effects of MCTP and the decline in ACE activity occurred coincident with the decrease in monolayer cellularity and appearance of marked cytotoxicity. 5. We conclude that high concentrations of MCTP decrease endothelial ACE activity. The decline in ACE activity is delayed and the magnitude and duration of the decrease corresponds to the degree ofMCTP-induced cytotoxicity. This suggests that altered endothelial ACE activity is unlikely to be a direct effect of MCTP on the enzyme but may reflect the delayed cell injury which results from exposure to this compound. PMID- 8242235 TI - Effect of capsazepine on the release of calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) induced by low pH, capsaicin and potassium in rat soleus muscle. AB - 1. We have determined the effect of the competitive antagonist capsazepine at the capsaicin receptor on the release of calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) from rat isolated soleus muscle induced by capsaicin (1 microM), by superfusion with low pH medium (pH 5) or by KCl (80 mM). 2. Each one of the three stimuli tested produced a marked CGRP-LI release. Total evoked release (fmol g-1) was 482 +/- 69, 169 +/- 20 and 253 +/- 43 for capsiacin, low pH medium and KCL, respectively. 3. Prior application of capsiacin (10 microM for 30 min followed by 30 min of washout) to produce capasaicin desensitization in vitro abolished CGRP-LI release induced by the three stimuli. 4. Capsazepine (1 100 microM, 45 min preincubation) inhibited the evoked CGRP-LI release. Capsaicin induced release was significantly inhibited by 77, 92 and 96% with 10, 30 and 100 microM capsazepine, respectively. Low pH-induced release was inhibited by 78, 84, 88 and 93% with 3, 10, 30 and 100 microM capsazepine, respectively. KCl-induced release was significantly inhibited by 55 and 93% with 30 and 100 microM (but not with 10 microM) capsazepine, respectively. 5. These findings demonstrate that capsazepine prevents low pH- and capsaicin-induced CGRP-LI release from rat soleus muscle at concentrations which do not affect the release evoked by KCl. These findings imply a relationship between the action of low pH and activation of the capsaicin receptor. At high concentrations, capsazepine produces a nonspecific inhibitory effect on CGRP-LI release from peripheral endings of the capsaicin-sensitive primary afferent neurone. PMID- 8242236 TI - Enhancement by calcitonin gene-related peptide of nicotinic receptor-operated noncontractile Ca2+ mobilization at the mouse neuromuscular junction. AB - 1. The involvement of calcitonin gene-related peptide (CGRP) in the mechanism of nicotinic acetylcholine receptor-operated noncontractile Ca2+ mobilization (not accompanied by twitch tension) was investigated by measuring Ca(2+)-aequorin luminescence at the neuromuscular junction of mouse diaphragm muscle treated with neostigmine. 2. Noncontractile Ca2+ transients were enhanced by 4-aminopyridine (100 microM), a K+ channel blocker, and inhibited by botulinum toxin (1-100 micrograms, i.p.) and hexamethonium (10-100 microM), a neuronal nicotinic receptor antagonist. 3. Noncontractile Ca2+ transients were diminished by CGRP8 37 (10-20 microM), a CGRP antagonist. CGRP (0.3-10 nM) prolonged the duration of noncontractile Ca2+ transients. The effect of CGRP was suppressed by CGRP8-37 (0.1 microM). 4. Noncontractile Ca2+ transients were inhibited by H-89 (0.1-1 microM), a protein kinase-A inhibitor. The catalytic subunit of protein kinase-A and AA373 (300 microM), a protein kinase-A activator, prolonged the duration of noncontractile transients. The prolongations either by CGRP or by AA373 were not observed in the presence of H-89 (0.1 microM). 5. Contractile (accompanied by twitch tension) but not noncontractile Ca2+ transients were decreased by 12-O tetradecanoyl phorbol 13-acetate (TPA, 0.3-1 microM), a protein kinase-C activator. Phospholipase A2 increased only contractile Ca2+ transients. Calmodulin-related agents affected neither type of Ca2+ transients. 6. These results provide the first evidence that nicotinic acetylcholine receptor-operated noncontractile Ca2+ mobilization is promoted by nerve-released CGRP activating protein kinase-A, and is dependent on the accumulated amounts of acetylcholine at the neuromuscular junction where desensitization might readily develop. PMID- 8242237 TI - Partial agonist effect of the platelet-activating factor receptor antagonists, WEB 2086 and WEB 2170, in the rat perfused heart. AB - 1. WEB 2086 and WEB 2170 are potent platelet-activating factor (PAF) receptor antagonists and have been used widely as pharmacological tools to investigate the actions of PAF in a variety of biological systems. 2. Low concentrations of WEB 2086 and WEB 2170 blocked the vasoconstrictor action of PAF in the rat perfused heart. In this study, we observed that moderate concentrations of WEB 2086 and WEB 2170 increased the perfusion pressure in rat isolated hearts under constant flow perfusion. The vasoconstrictor actions of WEB 2086 and WEB 2170 were not observed with a structurally different PAF receptor antagonist, FR-900452. 3. To determine whether this vasoconstrictor action of WEB 2086 involved non-specific effects or was via the activation of PAF receptors, hearts were pretreated with 1000 pmol PAF or 50 microM FR-900452. These pretreatments attenuated the vasoconstrictor action of 1 microM WEB 2086, suggesting that the action of WEB 2086 may be mediated via PAF receptors. Pretreatment with the leukotriene receptor antagonist (L-649,923, 5 microM) and the leukotriene synthesis inhibitor (MK-886, 10 microM) that are known to block the vasoconstrictor action of PAF receptor activation also attenuated the vasoconstrictor action of WEB 2086. Pretreatment with PAF or MK-886 attenuated the vasoconstrictor action of 0.5 microM WEB 2170. 4. When PAF receptors were activated by PAF in the perfused heart, significant amounts of leukotriene C4 and leukotriene C4/D4/E4 were detected in the coronary effluent. However, no significant amount of these leukotrienes was detected in the coronary effluent when hearts were perfused with 1 microM WEB 2086 or 0.5 microM WEB 2170. 5. In summary, our results indicate that WEB 2086 and WEB 2170 possess partial agonist effects in the rat perfused heart where they produced vasoconstriction via the activation of PAF receptor. This action could be attenuated by PAF pretreatment or a PAF receptor antagonist. The vasoconstrictor action of WEB 2086 and WEB 2170 involved the production of leukotrienes. But unlike the vasoconstrictor action of PAF, no significant amount of leukotrienes was detected in the effluent suggesting that the vasoconstrictor action of WEB 2086 and WEB 2170 may be explained on the basis of intracellularly or locally produced leukotrienes. PMID- 8242238 TI - Differential effects of acetylcholine, nitric oxide and levcromakalim on smooth muscle membrane potential and tone in the rabbit basilar artery. AB - 1. Endothelium-dependent hyperpolarization of smooth muscle cells in isolated, pre-contracted segments of rabbit basilar artery in response to acetylcholine (100 microM) was abolished in the presence of glibenclamide (10 microM). 2. Acetylcholine-evoked relaxation was unaffected by either glibenclamide or 65 mM potassium chloride, indicating that the change in membrane potential did not form an essential component of relaxation and that high concentrations of potassium did not inhibit the release or action of endothelium-derived relaxing factor in this vessel. 3. Saturated solutions of nitric oxide (NO) gas in solution (150 microM), which evoked maximal relaxation of arterial segments pre-contracted and depolarized by noradrenaline (10-100 microM), did not alter the membrane potential of either unstimulated or depolarized smooth muscle cells. 4. The potassium channel opener levcromakalim, evoked concentration-dependent relaxation and hyperpolarization in pre-constricted smooth muscle cells. The threshold concentrations for hyperpolarization and relaxation, the EC50 values and the maximally effective concentration of levcromakalim (around 30 nM, 150 nM and 10 microM, respectively) were not significantly different, and both components of the response were inhibited by glibenclamide (10 microM), indicating a close coupling between the two responses. 5. In the presence of 65 mM potassium chloride, the hyperpolarization to levcromakalim was abolished, while a small relaxation (25 +/- 4%) persisted, indicating an additional mechanism for relaxation to this agent. 6. These results show that different mechanisms underlie the relaxant action of potassium channel openers, NO and endothelium derived factors in cerebral arteries and provide further evidence that in the basilar artery, in contrast to some other vessels, endothelium-dependent hyperpolarization to acetylcholine is not important for smooth muscle relaxation. PMID- 8242239 TI - Bradykinin-induced airflow obstruction and airway plasma exudation: effects of drugs that inhibit acetylcholine, thromboxane A2 or leukotrienes. AB - 1. The mechanisms behind bradykinin-induced effects in the airways are considered to be largely indirect. The role of cholinergic nerves and eicosanoids, and their relationship in these mechanisms were investigated in guinea-pigs. 2. The role of cholinergic nerves was studied in animals given atropine (1 mg kg-1, i.v.), hexamethonium (2 mg kg-1, i.v.), or vagotomized. To study the role of eicosanoids, animals were pretreated with a thromboxane A2 (TxA2) receptor antagonist (ICI 192,605; 10(-6) mol kg-1, i.v.) or with a leukotriene (LT) receptor C4/D4/E4 antagonist (ICI 198,615; 10(-6) mol kg-1, i.v.). 3. After pretreatment with a drug, bradykinin (150 nmol) was instilled into the tracheal lumen. We measured both airway insufflation pressure (Pi), to assess airway narrowing, and the content of Evans blue dye in airway tissue, to assess plasma exudation. 4. Bradykinin instillation into the trachea caused an increase in Pi and extravasation of Evans blue dye. The increase in Pi was significantly attenuated by atropine or the TxA2 receptor antagonist, but not by hexamethonium, vagotomy or the LT receptor antagonist. 5. The bradykinin-induced exudation of Evans blue dye was significantly attenuated in the intrapulmonary airways by the TxA2 receptor antagonist, but not by atropine, hexamethonium, cervical vagotomy or the LT receptor antagonist. 6. A thromboxane-mimetic U-46619 (20 nmol kg-1, i.v. or 10 nmol intratracheally), caused both an increase in Pi and extravasation of Evans blue dye at all airway levels. Atropine pretreatment slightly attenuated the peak Pi after the intratracheal administration of U-46619, but not after i.v. administration. 7. We conclude that peripheral cholinergic nerves are involved in bradykinin-induced airflow obstruction but not plasma exudation, and that TxA2 is involved in both airflow obstruction and airway plasma exudation induced by bradykinin given via the airway route. TxA2-induced airflow obstruction is mediated only to a minor degree, via the release of acetylcholine in the airways. PMID- 8242240 TI - Differential affinity of dihydroimidazoquinoxalines and diimidazoquinazolines to the alpha 1 beta 2 gamma 2 and alpha 6 beta 2 gamma 2 subtypes of cloned GABAA receptors. AB - 1. In this study, we compared two series of newly discovered ligands for their selectivity to benzodiazepine sites in the alpha 1 beta 2 gamma 2 and the alpha 6 beta 2 gamma 2 subtypes of cloned gamma-aminobutyric acidA (GABAA) receptors, the latter being unique in not interacting with classical benzodiazepines. 2. The prototype compounds, U-85575 (12-chloro-5-(5-cyclopropyl-1',2',4'- oxadiazol-3' yl)-2,3-dihydro-diimidazo [1,5-a;1,2-c]quinazoline), and U-92330 (5-acetyl-3-(5' cyclopropyl-1',2',4'-oxadiazole-3'-yl)-7-chloro-4,5-d ihy dro [1,5 a]quinoxaline), appear to share an overlapping recognition site with classical benzodiazepines on the GABAA receptor, because their potentiation of GABA mediated Cl- currents in both subtypes were sensitive to Ro 15-1788, a classical benzodiazepine antagonist. 3. Minor changes in the ring substituents of the drugs reduced their affinity to the alpha 6 beta 2 gamma 2 subtype more pronouncedly than to the alpha 1 beta 2 gamma 2 subtype. The diimidazoquinazoline containing a 2-methyl group which projected below the plane of the rigid ring showed a markedly lower affinity to the alpha 6 beta 2 gamma 2 subtype as compared to its stereoisomer having the methyl group above the plane of the ring. Also, the dihydroimidazoquinoxalines containing the 5-benzoyl group showed a lower affinity to the alpha 6 beta 2 gamma 2 subtype than the 5-acetyl counterpart. In particular, the 5-benzoyl analogue containing a 6-fluoro group showed no interaction with the alpha 6 beta 2 gamma 2 subtype even at the concentration of 10 microM, probably due to stabilization of the benzoyl group in the out-of-plane region by the steric and electrostatic effects of the 6-fluoro group.4. We propose that the benzodiazepine site of the alpha 6 beta 2 gamma 2 subtype shares overlapping regions with that of the alpha 1 beta 2 gamma 2 subtype, but has a sterically restricted out-of-plane region, which may be also incompatible with the 5-phenyl group of classical benzodiazepines. PMID- 8242241 TI - Characterization of receptors for endothelins in the perfused arterial and venous mesenteric vasculatures of the rat. AB - 1. Endothelin-1 and -3 induced marked arterial and venous constrictions in the perfused mesenteric vasculature of the rat with endothelin-3 being at least 20 times less active than endothelin-1, on both arterial and venous sides of the vasculature. 2. Two ETB selective agonists, BQ-3020 and IRL 1620 (500 pmol), induced weak constrictions of the venous mesenteric vasculature and were inactive in the arterial side at doses up to 1000 pmol. 3. In mesenteric vasculatures precontracted with either methoxamine (arterial side) or the thromboxane A2 mimetic, U46619 (venous side), acetylcholine or bradykinin produced vasodilations of both arterial and venous vessels, whereas endothelin-3 induced vasodilations only on the arterial side. 4. A selective ETA receptor antagonist, BQ-123, blocked, in a concentration-dependent and reversible fashion, the vasoconstrictions induced by endothelin-1 on both sides of the mesenteric circulation (IC50; arterial side: 0.013 microM; venous side: 0.032 microM). 5. In contrast, the vasodilator responses induced by endothelin-3 on the arterial side of the precontracted mesenteric vasculature were not affected by BQ-123. 6. The present study illustrates the presence of ETA receptors which are responsible for vasoconstriction by endothelins in the arterial and venous mesenteric vasculatures. Furthermore, we suggest that the vasodilations induced by endothelin-3 in the arterial vasculature uniquely, are ETB receptor-mediated. PMID- 8242242 TI - Modulation of adjuvant arthritis by endogenous nitric oxide. AB - 1. The role of endogenous nitric oxide (NO) in adjuvant arthritis in Lewis rats has been studied by use of L-arginine, the amino acid from which NO is synthesized, and NG-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthase. Prolonged modulation (35 days) of the L-arginine: NO pathway in rats was achieved by dissolving test compounds in the drinking water (L-arginine: 3, 10 and 30 mg ml-1; L-NAME: 0.1, 1 and 10 mg ml-1). 2. Arthritis was exacerbated by L-arginine and suppressed by L-NAME in a dose-related fashion. Combined treatment with L-NAME (1 mg ml-1) and L-arginine (30 mg ml-1) did not modify the arthritis. 3. Reduced weight gain, which is a feature of adjuvant arthritis, was modified by these compounds so that L-arginine reduced weight gain whereas L-NAME increased weight gain compared with that in control animals. 4. D-Arginine (30 mg ml-1), NG-nitro-D-arginine methyl ester (D-NAME: 1 mg ml-1) and L-lysine (30 mg ml-1), an amino acid not involved in the generation of NO, were without effect on either arthritis or body weight gain. 5. Antigen-stimulated proliferation of T lymphocytes as well as generation of nitrite (NO2-) and release of acid phosphatase from macrophages were all enhanced in L-arginine-treated arthritic rats and reduced in L-NAME-treated animals. 6. These results suggest that endogenous NO modulates adjuvant arthritis, possibly by interfering with the activation of T-lymphocytes and/or macrophages. PMID- 8242243 TI - Protection by oestradiol against the development of cardiovascular changes associated with monocrotaline pulmonary hypertension in rats. AB - 1. We studied the effects of oestradiol 17 beta on the development of pulmonary vascular changes and right ventricular (RV) hypertrophy in response to monocrotaline in male Sprague-Dawley rats. 2. Rats were treated with either placebo or oestradiol 17 beta (10 mg) in the form of slow release pellets implanted subcutaneously 48 h before monocrotaline administration. Rats were injected with either saline or a single dose of monocrotaline (60 mg kg-1, i.m.). Pulmonary vascular changes and RV hypertrophy were studied at 4 weeks following monocrotaline administration. 3. Monocrotaline induced a significant increase in the ratio of right ventricle (RV) to left ventricle-plus-septum (LV + S) weights. Monocrotaline-treated rats also showed significant myointimal proliferation in small pulmonary arteries, decrease of arterial numbers and increase in the number of abnormal alveolar macrophages. 4. Oestradiol 17 beta attenuated myointimal hyperplasia in pulmonary vessels, decreased the RV/(LV + S) ratio in monocrotaline-treated rats. Oestradiol 17 beta had no significant effect on control animals. 5. Oestradiol treatment prevented the increase in lung wet to dry weight ratio, observed 7 days post monocrotaline administration. 6. These results suggest that oestradiol 17 beta protects against the pulmonary vascular remodelling and RV hypertrophy associated with monocrotaline-induced pulmonary hypertension in the rat. Oestradiol also protects against microvascular leak observed in the early days of lesion. PMID- 8242244 TI - Repeated administration of desipramine and a GABAB receptor antagonist, CGP 36742, discretely up-regulates GABAB receptor binding sites in rat frontal cortex. AB - 1. GABAB receptor binding site densities within laminar regions of the rat frontal cortex were examined autoradiographically following repeated administration (21 days) of the antidepressants desipramine, paroxetine and amitriptyline in addition to the GABAB receptor antagonists, CGP 35348 and CGP 36742. beta 1-Adrenoceptor autoradiography was studied in parallel with that for GABAB receptor sites. 2. The effects of these compounds were examined concomitantly on the GABAB receptor-mediated inhibition of forskolin- and enhancement of noradrenaline-stimulated cyclic AMP production. 3. GABAB receptor binding was increased by both desipramine (20 mg kg-1, p.o. and 10 mg kg-1, i.p.) and CGP 36742 (100 mg kg-1, i.p.) in the outer laminar region of the frontal cortex by around 50% above control levels. Conversely, no significant changes were mediated by paroxetine, amitriptyline, CGP 35348 or the GABAB receptor agonist, baclofen. 4. With the exception of paroxetine, all compounds down regulated the total beta-adrenoceptor population throughout frontal cortical laminae which was attributable to the beta 1-adrenoceptor subtype. In contrast, the reduction in beta-adrenoceptors mediated by CGP 35348 and CGP 36742 did not occur as a consequence of reduced beta 1-adrenoceptor numbers. 5. Protracted treatment with CGP 35348, failed to influence forskolin-stimulated cyclic AMP production; however, a significant increase in the accumulation of cyclic AMP produced in response to forskolin was seen after treatment with CGP 36742. 6. Such discretely localized changes in GABAB receptor densities induced by desipramine and CGP 36742 may provide an explanation for the discrepancies reported in membrane binding studies and possibly implicate a role for GABAB receptor antagonists in antidepressant therapy. PMID- 8242245 TI - Differential inhibition by NG-monomethyl-L-arginine of vasodilator effects of acetylcholine and methacholine in human forearm vasculature. AB - 1. We compared the effects of NG-monomethyl-L-arginine (L-NMMA), an NO synthase inhibitor, on vasodilatation produced by acetylcholine and methacholine in human forearm vasculature. 2. Acetylcholine (83 nmol min-1) infused into the brachial artery of 8 healthy volunteers caused a submaximal increase in forearm blood flow, measured by venous occlusion plethysmography, from 3.3 +/- 0.5 (mean +/- s.e. mean) to 13.3 +/- 1.7 ml min-1 100 ml-1. 3. Co-infusion of L-NMMA (4 mumol min-1) with acetylcholine (83 nmol min-1) over 6 min resulted in a 58% +/- 12% fall in the response to acetylcholine whereas during co-infusion of saline over the same time period in the same subjects (n = 8) on a different day the response to acetylcholine fell by only 9% +/- 17% (P < 0.01). 4. Methacholine (1.5 and 15 nmol min-1) increased forearm blood flow from 2.5 +/- 0.4 to 5.9 +/- 0.9 and from 3.2 +/- 0.4 to 17.0 +/- 1.9 ml min-1 100 ml-1 respectively. 5. Co-infusion of L NMMA (4 mumol min-1) had no significant effect on the response to methacholine (1.5 or 15 nmol min-1) when compared with saline control (n = 8). Co-infusion of a higher dose of L-NMMA (8 mumol min-1) with methacholine (1.5 nmol min-1) did not significantly inhibit the vasodilator response (n = 7). 6. These results suggest that, in human forearm vasculature, methacholine acts predominantly through mechanisms other than the L-arginine/nitric oxide pathway. PMID- 8242246 TI - Pulmonary effects of endogenous and exogenous nitric oxide in the pig: relation to cigarette smoke inhalation. AB - 1. Pentobarbitone-anaesthetized pigs were challenged with cigarette smoke (unfiltered or filtered through a Cambridge glass fibre filter to remove the particulate phase including nicotine), as well as nicotine aerosol and the gas phase components nitric oxide (NO) and carbon monoxide (CO); the effects on the bronchial and pulmonary circulations, and pulmonary airway mechanics, were examined. The relative importance of endogenous NO mechanisms in the pig lung was also studied by giving the NO synthesis inhibitor NG-nitro-L-arginine (L-NOARG; 50 mg kg-1) intravenously. Mean arterial pressure and blood flow in the bronchial, pulmonary and femoral circulations were measured, the latter with ultrasonic flow probes around the supplying arteries, and vascular resistance (VR) was calculated. Changes in pulmonary airways resistance (Rpulm) and lung dynamic compliance (Cdyn) were also determined. Finally, the concentration of NO in inhaled air during cigarette smoke and NO gas challenges was continuously monitored by a chemiluminescence method and the relative contribution of NO in cigarette smoke-induced vascular effects in the pig lung was calculated. 2. Cigarette smoke challenge, with or without a Cambridge filter, caused a rapid vasodilator response in the bronchial circulation and the major part (75%) of this response was probably caused by NO present in smoke. NO challenge caused profound bronchial vasodilation with dose-response characteristics between 10 and 100 p.p.m. The small part of the cigarette smoke-induced response not explained by the NO content may be caused by CO, showing weak vasodilator effect in the bronchial circulation. The L-NOARG-induced relative increase in bronchial VR was 2-3 times higher than the changes in pulmonary, femoral and systemic VR, suggesting a strong influence of endothelial NO mechanisms on basal tone in the bronchial circulation.3. Challenge with unfiltered cigarette smoke induced variable responses in the pulmonary circulation,whereas inhalation of filtered smoke caused consistent pulmonary vasodilatation. The major part of this vasodilator response was probably caused by NO, which was a potent dilator of the pulmonary circulation with maximal effect achieved with as little as 10 p.p.m. The effect of NO may be opposed in unfiltered smoke by the particulate phase (but not nicotine), presumably by inducing sympathetic reflexes. L-NOARG caused similar relative increases in pulmonary, femoral and systemic VR.4. Cigarette smoke inhalation induced bronchodilatation in the pentobarbitone-anaesthetized pig as revealed by changes in Rpulm and Cdyn. Both NO and nicotine may contribute to this response. NO inhalation reduced Rpulm in the basal state with maximal effect at 30 p.p.m. The mechanism for NO-induced bronchodilatation may be indirect in the pig, since pretreatment with L-NOARG blocked the response. L NOARG did not affect basal Rpulm.5. In conclusion, bronchial vasodilatation caused by continuous cigarette smoke inhalation in the pig,seems to be largely mediated (approximately 75%) by NO. The remaining part could be mediated by CO. Cigarette smoke particles, but not nicotine, may counteract NO-induced relaxation in the pulmonary circulation, thus resulting in variable effects in the pulmonary circulation during challenge with unfiltered cigarette smoke. NO also acts as a bronchodilator in the pig, but the mechanism may be indirect. Finally, endogenous NO mechanisms appear to be strongly involved in the control of basal tone in the bronchial circulation, less so in the pulmonary circulation and not at all in bronchial smooth muscle. PMID- 8242247 TI - Activation of the phospholipase C pathway by ATP is mediated exclusively through nucleotide type P2-purinoceptors in C2C12 myotubes. AB - 1. The presence of a nucleotide receptor and a discrete ATP-sensitive receptor on C2C12 myotubes has been shown by electrophysiological experiments. In this study, the ATP-sensitive receptors of C2C12 myotubes were further characterized by measuring the formation of inositol(1,4,5)trisphosphate (Ins(1,4,5)P3) and internal Ca2+. 2. The nucleotides ATP and UTP caused a concentration-dependent increase in Ins(1,4,5)P3 content with comparable time courses (EC50: ATP 33 +/- 2 microM, UTP 80 +/- 4 microM). ADP was less effective in increasing Ins(1,4,5)P3 content of the cells, while selective agonists for P1-, P2X- and P2Y purinoceptors, adenosine, alpha,beta-methylene ATP and 2-methylthio ATP, appeared to be ineffective. 3. Under Ca(2+)-free conditions, the basal level of Ins(1,4,5)P3 was lower than in the presence of Ca2+, and the ATP- and UTP-induced formation of Ins(1,4,5)P3 was diminished. 4. The Ins(1,4,5)P3 formation induced by optimal ATP and UTP concentrations was not additive. ATP- and UTP-induced Ins(1,4,5)P3 formation showed cross-desensitization, whereas cross desensitization was absent in responses elicited by one of the nucleotides and bradykinin. 5. The change in Ins(1,4,5)P3 content induced by effective nucleotides was inhibited by suramin. Schild plots for suramin inhibition of Ins(1,4,5)P3 formation in ATP- and UTP-stimulated myotubes showed slopes greater than unity (1.63 +/- 0.09 and 1.37 +/- 0.11, respectively). Apparent pA2 values were 4.50 +/- 0.48 and 4.41 +/- 0.63 for ATP and UTP, respectively. 6. Stimulation of the cells with ATP or UTP induced a rapid increase in intracellular Ca2+, followed by a slow decline to basal levels. Ca2+ responses reached lower maximal values and did not show the slow phase in the absence of extracellular Ca2+. The ATP and UTP-evoked increase in intracellular Ca2+ was not additive and showed cross-desensitization. Cross-desensitization was absent in myotubes stimulated with one of the nucleotides and bradykinin.7. These results show that ATP- and UTP-induced formation of Ins(1,4,5)P3, Ca2+ release from internal stores and Ca2+-influx from the extracellular space are mediated exclusively via the nucleotide type P2-purinoceptor in mouse C2C12 myotubes. PMID- 8242248 TI - Similar effects of ethanol and flumazenil on acquisition of a shuttle-box avoidance response during withdrawal from chronic ethanol treatment. AB - 1. Acquisition of a two-way shuttle-box avoidance response is facilitated by ethanol. This facilitated acquisition of an avoidance response to ethanol was attenuated during withdrawal from chronic-ethanol diet intake (i.e. tolerance developed by ethanol). The deficit in the avoidance task after chronic ethanol treatment could be overcome by increasing the dose of ethanol. 2. Flumazenil, a benzodiazepine antagonist, also facilitated acquisition of the avoidance response in control rats. This response to flumazenil was significantly reduced during withdrawal from chronic-ethanol treatment. This reduced avoidance responding during withdrawal also could be overcome by increasing the dose of flumazenil. 3. The benzodiazepine-inverse agonist, RO 15-4513, produced a deficit in avoidance responding that was antagonized by both ethanol and flumazenil in a dose-related manner. 4. To determine whether flumazenil has the properties of a benzodiazepine agonist, it was established that, unlike the benzodiazepine chlordiazepoxide, flumazenil did not enhance the ethanol-induced deficit in the aerial righting reflex. Additionally, flumazenil blocked the action of chlordiazepoxide in this procedure, consistent with the benzodiazepine antagonist action of flumazenil. 5. Data collected are consistent with the hypothesis that an endogenous substance with the properties of a benzodiazepine-inverse agonist antagonizes the anticonflict actions of acutely administered ethanol during withdrawal from chronic-ethanol exposure. PMID- 8242249 TI - Pharmacological profile of valsartan: a potent, orally active, nonpeptide antagonist of the angiotensin II AT1-receptor subtype. AB - 1. The pharmacological profile of valsartan, (S)-N-valeryl-N-([2'-(1H-tetrazol-5 yl)biphenyl-4-yl]-methyl)-vali ne, a potent, highly selective, and orally active antagonist at the angiotensin II (AII) AT1-receptor, was studied in vitro and in vivo. 2. Valsartan competed with [125I]-AII at its specific binding sites in rat aortic smooth muscle cell membranes (AT1-receptor subtype) with a Ki of 2.38 nM, but was about 30,000 times less active in human myometrial membranes (AT2 receptor subtype). 3. In rabbit aortic rings incubated for 5 min with valsartan, at concentrations of 2, 20 and 200 nM, the concentration-response curve of AII was displaced to the right and the maximum response was reduced by 33%, 36% and 40%, respectively. Prolongation of the incubation time with valsartan to 1 h or 3 h, further reduced the maximum response by 48% or 59% (after 20 nM) and by 59% or 60% (after 200 nM) respectively. After 3 h incubation an apparent pKb value of 9.26 was calculated. Contractions induced by noradrenaline, 5-hydroxytryptamine, or potassium chloride were not affected by valsartan. No agonistic effects were observed in the rabbit aorta at concentrations of valsartan up to 2 microM. 4. In bovine adrenal glomerulosa, valsartan inhibited AII-stimulated aldosterone release without affecting the maximum response (pA2 8.4). 5. In the pithed rat, oral administration of valsartan (10 mg kg-1) shifted the AII-induced pressor response curves to the right, without affecting responses induced by the electrical stimulation of the sympathetic outflow or by noradrenaline. Animals treated with valsartan 24 h before pithing also showed significant inhibition of the response to AII. 6. In conscious, two-kidney, one-clip renal hypertensive rats (2K1C), valsartan decreased blood pressure in a dose-dependent manner after single i.v. or oral administration. The respective ED30 values were 0.06 mg kg-1 (i.v.) and 1.4 mg kg-1 (p.o.). The antihypertensive effect lasted for at least 24 h after either route of administration. After repeated oral administration for 4 days (3 and 10 mg kg-1 daily), in 2K1C renal hypertensive rats, systolic blood pressure was consistently decreased, but heart rate was not significantly affected. 7. In conscious, normotensive, sodium-depleted marmosets, valsartan decreased mean arterial pressure, measured by telemetry, after oral doses of 1-30 mg kg-1. The hypotensive effect persisted up to 12 h after 3 and 10 mg kg-1 and up to 24 h after 30 mg kg-1. 8. In sodium-depleted marmosets, the hypotensive effect of valsartan lasted longer than that of losartan(DuP 753). In renal hypertensive rats, both agents had a similar duration (24 h), but a different onset of action (valsartan at 1 h, losartan between 2 h and 24 h).9. These results demonstrate that valsartan is a potent, specific, highly selective antagonist of AII at theAT1-receptor subtype and does not possess agonistic activity. Furthermore, it is an efficacious, orally active, blood pressure lowering agent in conscious renal hypertensive rats and in conscious normotensive,sodium-depleted primates. PMID- 8242250 TI - Effect of a calcitonin gene-related peptide antagonist (CGRP8-37) on skin vasodilatation and oedema induced by stimulation of the rat saphenous nerve. AB - 1. The effect of the calcitonin gene-related peptide antagonist (CGRP8-37, 400 nmol kg-1, i.v.) on the increased blood flow induced by calcitonin gene related peptide (CGRP), vasodilator prostaglandins, and topical capsaicin was measured with a laser Doppler blood flow meter in rat abdominal skin. 2. The saphenous nerve was electrically stimulated and the effect of CGRP8-37 (400 nmol kg-1, i.v.) on the increased blood flow (measured by laser Doppler flowmetry) and oedema formation (measured by the extravascular accumulation of [125I]-albumin) was investigated in the rat hind paw. 3. CGRP8-37 (400 nmol kg-1, i.v.) had no effect on basal cutaneous blood flow at uninjected sites and sites injected with Tyrode buffer, but acted selectively to inhibit the increased blood flow induced by intradermal CGRP (10 pmol/site, P < 0.05), but not that induced by prostaglandin E2 (PGE2, 300 pmol/site) or carba-prostacyclin (cPGI2, 100 pmol/site). 4. Capsaicin (0.1-33 mM), applied topically, acted in a dose-related manner to increase blood flow. CGRP8-37 (400 nmol kg-1, i.v.) almost totally inhibited blood flow induced by capsaicin (10 mM; P < 0.05) but did not significantly inhibit blood flow induced by a higher dose of capsaicin (33 mM). 5. The increased blood flow induced by short stimulation of the saphenous nerve (10 V, 1 ms, 2 Hz for 30 s) was inhibited by 76%, 5 min after i.v. CGRP8-37 (400 nmol kg-1, i.v., P < 0.05). 6. A longer (5 min) electrical stimulation of the saphenous nerve caused oedema formation, in addition to increased blood flow. The oedema formation was significantly inhibited by CGRP8-37 (400 nmol kg-1,i.v., P<0.05).7. The results suggest that the potent microvascular vasodilator neuropeptide, CGRP, is responsible for the increased blood flow observed after short stimulation of the saphenous nerve and that endogenous CGRP contributes in a pro-inflammatory manner to neurogenic oedema formation in the rat hind paw. PMID- 8242251 TI - Use of the endothelin antagonists BQ-123 and PD 142893 to reveal three endothelin receptors mediating smooth muscle contraction and the release of EDRF. AB - 1. We have compared the receptors mediating the contractions of rings of rat thoracic aorta or rabbit pulmonary artery and rat stomach strips in response to the endothelin/sarafotoxin (ET/SX) family of peptides and to those mediating endothelium-dependent vasodilations within the isolated perfused mesentery of the rat. To discriminate ETA receptors from ETB receptors we have used the criteria that ET-1 is more active than SX6c on ETA receptors, and that the ET/SX peptides are equiactive on ETB receptors. We have also assessed the effects of the ETA receptor-selective antagonist BQ-123, and the non-selective ET receptor antagonist PD 142893 on the responses of each preparation to the ET/SX peptides. 2. ET-1-induced constrictions of the rat thoracic aorta (EC50 3 x 10(-10) M), a prototypic ETA receptor-mediated response, or isolated perfused mesentery of the rat were antagonized by BQ-123 (10(-5) M) or PD 142893 (10(-5) M). SX6c did not constrict either the rat isolated perfused mesentery or the rat thoracic aorta. Thus, ETA receptors mediate these constrictions. 3. ET-1 and SX6c were approximately equipotent in constricting rabbit pulmonary artery rings (EC50S 3-6 x 10(-10) M). Neither BQ-123 (10(-5) M) nor PD 142893 antagonized the contractions induced by ET-1. These effects suggest mediation by ETB receptors but PD 142893 (10(-5) M) did give a 3 fold antagonism of constrictions induced by SX6c. 4. SX6c was more potent than ET-1 in contracting the rat stomach strip (threshold concentrations 10(-10) and 3 x 10(-10) M). Contractions to ET-1 or SX6c were unaffected by BQ-123 (10-5 M), again indicative of ETB receptor mediated events. PD 142893 (10-5 M) was ineffective against ET-1 but produced a 3 fold antagonism of SX6c.5. In the rat isolated perfused mesentery ET-1 or SX6c (0.3-300pmol) were equipotent in producing dose-related vasodilatations that were unaffected by BQ-123 (10-6 M), indicative of an ETB receptor mediated response. In contrast to the other ETB-mediated responses, PD 142893 (10-6 M) strongly antagonized these vasodilatations.6. Thus, ETA receptors mediate constrictions of the rat thoracic aorta and rat isolated perfused mesentery whereas ETB receptors mediate constrictions of the rabbit pulmonary artery and rat stomach strip and endothelium-dependent dilatations within the mesentery. However, within the group of ETB receptor-mediated responses, endothelium-dependent vasodilatations are sensitive to PD 142893, whereas contractions of the isolated smooth muscle preparations are not. Thus, the receptor present on the endothelium responsible for the release of nitric oxide in response to the ET/SX peptides is most probably different from that present on smooth muscle that mediates BQ-123 insensitive contractions. PMID- 8242252 TI - Characterization of endothelin receptors mediating the effects of the endothelin/sarafotoxin peptides on autonomic neurotransmission in the rat vas deferens and guinea-pig ileum. AB - 1. To characterize the receptors mediating the effects of the endothelin/sarafotoxin family of peptides on the responses to electrical stimulation of the rat vas deferens (RVD) and guinea-pig ileum (GPI) we have used endothelin-1 (ET-1), ET-3, sarafotoxin 6b (SX6b) and SX6c as agonists and the endothelin-receptor antagonists BQ-123 (ETA receptor selective) and PD 142893 (non-selective). 2. In the RVD, ET-1 and SX6b increased the twitches induced by field stimulation starting at a threshold concentration of 10(-10) M while the threshold concentration for ET-3 was 3 x 10(-9) M. SX6c (up to 3 x 10(-8) M) did not potentiate the twitches. SX6b produced significantly (P < 0.05) greater potentiations than ET-1 at concentrations of 3 x 10(-9) M and higher, and 10(-7) M ET-3 also produced a significantly greater effect than ET-1 at the same concentration. Thus, at threshold the rank order of peptides was ET-1 = SX6b > ET 3 >>> SX6c, and at concentrations of 3 x 10(-8) M and higher, SX6b > ET-3 > ET-1 >>> SX6c. 3. In the presence of BQ-123 or PD 142893 (10(-5) M) the threshold concentrations for ET-1 to augment the twitches were increased 30 fold. In the same conditions neither SX6b nor ET-3 potentiated the responses. The relative activities of the endothelin/sarafotoxin peptides and the effectiveness of the endothelin receptor antagonists are consistent with postjunctional ETA receptors mediating these effects. 4. In the transmurally stimulated GPI the endothelin/sarafotoxin peptides produced two effects; an increase in the basal tension of the tissues and an inhibition of the twitch responses. To increase the basal tension the peptides had the order of potency ET-1 > SX6b>> ET-3 = SX6c. These direct effects of ET-1 or SX6b were strongly antagonized (100 fold) by either BQ-123 (10-5M) or PD 142893(10-5 M). Thus, ETA receptors mediate contractions of the GPI induced by these peptides.5. The endothelin/sarafotoxin peptides were approximately equipotent at depressing twitches of the GPI in response to transmural stimulation (EC50s, 4 x 10-11 to 1.5 x 10-10 M). The depressions induced byET-1 were unaffected by either BQ-123 (10-5 M) or PD 142893 (10-5 M). BQ-123 produced a small(three fold) antagonism of the inhibitory effects of ET-3 or SX6c. These results indicate that a receptor of the ETB type mediates the inhibitory effects of the endothelin/sarafotoxin peptides on neurotransmission in the GPI.6. Thus, both ETA receptors and ETB receptors mediate the effects of the endothelin/sarafotoxinpeptides on neurotransmission. PMID- 8242253 TI - Tachykinin NK1 but not NK2 receptors mediate non-cholinergic excitatory junction potentials in the circular muscle of guinea-pig colon. AB - 1. The effect of tachykinin NK1 and NK2 receptor antagonists on noncholinergic excitatory junction potentials (e.j.ps) evoked by electric field stimulation (EFS) in the circular muscle of the guinea-pig proximal colon was investigated by means of a sucrose-gap technique. 2. In the presence of 1 microM atropine, submaximal EFS (10 Hz, 20-30 V, 0.5 ms pulse width, 1 s train duration) evoked an inhibitory junction potential (i.j.p.) followed by e.j.p. with superimposed action potentials (APs) and contraction. Addition of either NG-nitro-L-arginine (L-NOARG, 0.1 mM) or apamin (0.1 microM) inhibited the evoked i.j.p. and the combined administration of the two agents almost abolished it. In the presence of both L-NOARG and apamin, an atropine-resistant e.j.p. was the only electrical response evoked by EFS in 50% of cases and a small i.j.p. (10% of original amplitude) followed by e.j.p. was evident in the remainder. 3. In the presence of L-NOARG and apamin, the tachykinin NK1 receptor antagonists, (+/-)-CP 96,345 and GR 82,334 (10 nM-3 microM) concentration-dependently inhibited the atropine resistant e.j.p. and accompanying contraction evoked by EFS. EC50 values were: 0.77 microM (e.j.p. inhibition) and 0.22 microM (inhibition of contraction) for (+/-)-CP 96,345; 0.61 microM (e.j.p. inhibition) and 0.20 microM (inhibition of contraction) for GR 82,334. The tachykinin NK2 receptor antagonists, MEN 10,376 (up to 3 microM) and SR 48,968 (up to 1 microM) had no effect on the atropine resistant e.j.p. MEN 10,376 (3 microM) but not SR 48,968 produced a slight inhibition of the evoked contraction. 4. (+/- )-CP 96,345 (3 microM) and GR 82,334 (3 microM) markedly reduced (81 and 89% inhibition, respectively)the atropine-resistant ej.p. in the absence of L-NOARG and apamin, without affecting the ij.p. MEN 10,376 (3 microM) and SR 48,968 (1 microM) had no significant effect on noncholinergic ij.p. and ej.p. evoked in the absence of apamin and L NOARG.5. The electrical and mechanical responses to the NK, receptor agonist [Sar9]substance P (SP) sulfone were blocked by (+/-)-CP 96,345 (3 1M) or GR 82,334 (3 microM) which, at the same concentration, failed to affect the responses to the NK2 receptor agonist [PAla8] neurokinin A (NKA) (4-10). In contrast, MEN10,376 (3 microM) or SR 48,968 (1 microM) blocked the response to [beta Ala8]NKA(4-10) without affecting the response to [Sar9]SP sulfone.6. In the presence of L-NOARG and apamin, and in the absence of atropine, EFS of low pulse width(0.02-0.03 ms, other parameters as above) produced cholinergic ej.ps and contraction which were unaffected by GR 82,334 (3 microM). (+/-)-CP 96,345 (3 JAM) produced 24% reduction in the area of the atropine-sensitive ej.p. without affecting the peak amplitude of ej.p. or contraction.7. These findings demonstrate that the noncholinergic ej.ps and accompanying contraction of the circular muscle of the guinea-pig colon are produced through activation of intramural tachykininergic nerves and that the resultant smooth muscle response is almost entirely mediated through NK1 receptors. PMID- 8242254 TI - Muscarinic effect of atrial natriuretic peptide on rabbit airways. AB - 1. The aim of the present work was to investigate under which circumstances atrial natriuretic peptide (ANP) modulates airway resistance. 2. Of the six groups of rabbits (n = 5) studied, three received an infusion of ANP (80 ng min-1 kg-1 i.v.) for a period of 100 min, while the other three were infused with the vehicle. Before receiving the infusion of ANP or the vehicle, the animals were pretreated with atropine (0.5 mg kg-1 i.v.), propranolol (2 mg kg-1 i.v.) or not pretreated. After 75 min of infusion of ANP, bronchoconstriction was induced by inhalation of histamine. Respiratory resistance (Rrs) was measured before and 3, 5, 10, 15 and 20 min post-histamine challenge. 3. Following 75 min of ANP infusion, plasma ANP concentration increased from 153 +/- 52 (mean +/- s.e.mean) to 1441 +/- 203 pg ml-1 (P < 0.05) without affecting baseline Rrs. Control Rrs values (12.5-20.4 cmH2O l-1 s) were significantly increased following the inhalation of histamine (P < 0.001). By themselves, atropine, propranolol or ANP did not modify the histamine-induced increase in Rrs. However, when the animals were pretreated with atropine, ANP infusion significantly reduced the increase in Rrs induced by histamine (30 +/- 2 vs 51 +/- 6 cmH2O l-1 s; P < 0.05). 4. These data suggest that ANP has an indirect modulating effect on the airway smooth muscle and will decrease Rrs when muscarinic receptors are blocked. PMID- 8242255 TI - Characterization of the effects of lithium on phosphatidylinositol (PI) cycle activity in human muscarinic m1 receptor-transfected CHO cells. AB - 1. The effects of lithium on [3H]-inositol and [3H]-cytidine incorporation into [3H]-inositol monophosphates ([3H]-InsP1) and [3H]-cytidine monophosphorylphosphatidate ([3H]-CMP-PA), respectively, and inositol 1,4,5 trisphosphate (InsP3) and inositol 1,3,4,5-tetrakisphosphate (InsP4) mass were studied in carbachol-stimulated human m1 muscarinic receptor-transfected Chinese hamster ovary cells (m1 CHO cells). 2. Lithium alone (10 mM) had no appreciable effects on any of the four parameters measured; it was only in carbachol stimulated cells that the effects of lithium became apparent. 3. In the presence of carbachol (1 mM), lithium (10 mM) caused a relatively rapid (within 5 min) accumulation of [3H]-InsP1 and [3H]-CMP-PA which continued up to about 20-30 min, after which accumulation slowed down. On the other hand, the elevation in InsP3 and InsP4 levels produced by carbachol was not altered by lithium in the short term and only at later times (> 20-30 min) was the response attenuated, with InsP3 and InsP4 levels approaching basal. 4. The effects of lithium on carbachol stimulated [3H]-InsP1 and [3H]-CMP-PA accumulation and the attenuation of the carbachol-induced elevation of InsP3 and InsP4 were all dose-dependent, with EC50s in the region of 1 mM. 5. The lithium-induced effects on [3H]-CMP-PA and InsP3 and InsP4 in carbachol-stimulated cells could be reversed, in a dose dependent manner, by preincubation with exogenous myo-inositol (EC50 = 2-3 mM) but not by the inactive analogue scyllo-inositol, indicating that these effects occur as a consequence of depletion of inositol. 6. The temporal effects of lithium are consistent with lithium inhibiting inositol monophosphatase,causing accumulation of InsP1, resulting in lower free inositol levels. This leads to accumulation of CMP-PA and reduced PI synthesis which, once agonist-linked membrane inositol phospholipids are depleted, produces attenuated InsP3 and InsP4 responses.7. These results in ml CHO cells support the hypothesis that lithium affects the PI cycle cell signalling pathway by depletion of inositol due to inhibition of inositol monophosphatase. PMID- 8242256 TI - The role of nitric oxide in cholinergic neurotransmission in rat trachea. AB - 1. We have investigated the role of nitric oxide (NO) in cholinergic contraction in rat trachea. 2. Methylene blue (10 nM to 30 microM) potentiated cholinergic contraction induced by electrical field stimulation (EFS) at 5 Hz in a concentration-dependent fashion. At a concentration of 30 microM, methylene blue decreased responses to log EFS frequency, producing 50% of maximum contraction from a control value of 0.74 +/- 0.09 Hz to 0.30 +/- 0.05 Hz without a significant effect on concentration-response curves to acetylcholine (ACh). 3. NG monomethyl-L-arginine (L-NMMA; 100 microM) also potentiated cholinergic contraction induced by EFS at 5 Hz (131.5 +/- 4.6% of control) without having any effect against ACh (3 microM)-induced contractions. Likewise, L-NMMA (100 microM) significantly increased EFS (5 Hz)-evoked release of ACh from tracheal segments into the bath solution (51.4 +/- 4.0 pmol ml-1 in the presence of L-NMMA and 35.0 +/- 1.8 pmol ml-1 in the absence of L-NMMA, respectively). 4. Administration of NO (present in acidified solution of NaNO2) (1 nM to 10 microM) and sodium nitroprusside (100 nM to 10 microM) concentration-dependently reduced EFS (5 Hz) induced cholinergic contractions without having a significant effect on ACh (3 microM)-induced contractions. These results were unaffected by prior exposure of the tissues to L-NMMA (100 microM). 5. Dibutyryl cyclic GMP (3 mM) also reduced cholinergic contractions induced by EFS at 5 Hz (70.1 +/- 3.6% of control) without any significant effect on ACh (3 microM)-induced contractions. 6. Pretreatment of tissues with capsaicin (30 microM) or a-chymotrypsin (1 u ml-') failed to inhibit methylene blue (30 microM)-induced potentiation of responses to EFS at 5 Hz.7. These results suggest that an endogenous NO-like factor may mediate prejunctional inhibition of cholinergic contraction through a cyclic GMP dependent mechanism in rat trachea. PMID- 8242257 TI - The effects of a converting enzyme inhibitor (captopril) and angiotensin II on fetal renal function. AB - 1. Renal function was studied in chronically catheterized fetal sheep (119-128 days gestation), before and during treatment of the ewe with the angiotensin converting enzyme (ACE) inhibitor, captopril, which crosses the placenta and blocks the fetal renin angiotensin system. 2. An i.v. dose of 15 mg (about 319 micrograms kg-1) of captopril to salt-replete ewes followed by an infusion to the ewe of 6 mg h-1 (about 128 micrograms kg-1 h-1) caused a fall in fetal arterial pressure (P < 0.01), and a rise in fetal renal blood flow (RBF) from 67.9 +/- 5.6 to 84.9 +/- 8.3 ml min-1 (mean +/- s.e. mean) (P < 0.05). Renal vascular resistance and glomerular filtration rate (GFR) fell (P < 0.01); fetal urine flow (P < 0.01); fetal urine flow (P < 0.01) and sodium excretion declined (P < 0.05). 3. Ewes were treated for the next 2 days with 15 mg captopril twice daily. On the 4th day, 15 mg was given to the ewe and fetal renal function studied for 2 h during the infusion of captopril (6 mg h-1) to the ewe. Of the 9 surviving fetuses, 3 were anuric and 3 had low urine flow rates. When 6 micrograms kg-1 h-1 of angiotensin II was infused directly into the fetus RBF fell from 69 +/- 10.1 ml min-1 to 31 +/- 13.9 ml min-1, GFR rose (P < 0.05) and urine flow (P < 0.01) and sodium excretion increased in all fetuses. 4. It is concluded that the small fall in fetal arterial pressure partly contributed to the fall in fetal GFR but in addition, efferent arteriolar tone fell so that the filtration pressure fell further. Thus maintenance of fetal renal function depends on the integrity of the fetal renin angiotensin system. These findings explain why use of ACE inhibitors in human pregnancy is associated with neonatal anuria. PMID- 8242258 TI - Ionomycin-induced acetylcholine release and its inhibition by adenosine at frog motor nerve endings. AB - 1. Acetylcholine (ACh) evoked secretion by the calcium ionophore, ionomycin, was studied at frog motor nerve endings. 2. Bath application of ionomycin stimulated an irreversible increase in the rate of spontaneous, quantal ACh release in the presence of extracellular Ca2+. In contrast, local application of ionomycin stimulated a rapid, reversible acceleration of spontaneous ACh release. 3. The magnitude of the secretory response to ionomycin was dependent both upon the concentration of ionophore and the concentration of extracellular Ca2+. 4. Adenosine or 2-chloroadenosine inhibited ionomycin-stimulated ACh release with the same potency and efficacy observed previously for these adenosine analogues as inhibitors of ACh secretion evoked by nerve impulses. 5. These results support the conclusion that adenosine receptor activation inhibits quantal ACh secretion at a site distal to that of Ca2+ entry at frog motor nerve endings. PMID- 8242259 TI - Stimulated eosinophils and proteinases augment the transepithelial flux of albumin in bovine bronchial mucosa. AB - 1. The apical to basolateral transmucosal flux of albumin has been measured in isolated sheets of bovine bronchial and tracheal mucosa. Under resting conditions the net unidirectional flux in the bronchial mucosa was not significantly different from that measured previously for the basolateral to apical vector. In contrast, the apical to basolateral flux in the tracheal mucosa was significantly lower than that measured in the opposite direction. 2. Addition of guinea-pig peritoneal eosinophils to the apical side of the tissues had no significant effect on the transmucosal flux of albumin in either the bronchial or tracheal mucosa. 3. When eosinophils were stimulated with the ionophore A23187 or by opsonic adherence to tissues treated with a guinea-pig anti-bovine airway epithelium antibody, the bronchial mucosal sheets that had been exposed showed a significant increase in the transmucosal flux of albumin. However, tissues from the tracheal mucosa were resistant to the effects of stimulated eosinophils. 4. Histologically, sheets of mucosa from bovine main bronchi that had been exposed to stimulated eosinophils were characterized by epithelial injury consisting of loss of columnar epithelium from the underlying basal cell layer and biomatrix. Much less evidence of cellular injury was observed in tracheal tissues. 5. Bacterial collagenases applied to the apical side of the sheets were shown to increase the permeability of the bronchial mucosa to albumin and to produce histological changes that had similarities with the pattern of damage produced by stimulated eosinophils. 6. These observations demonstrate that the ability of eosinophils to injure the bronchial mucosa is independent of the side of the tissue on which they are present. Furthermore, key aspects of the injury process may be reproduced, at least in part, by metalloproteinases. PMID- 8242260 TI - Extracellular ATP and UTP activation of phospholipase D is mediated by protein kinase C-epsilon in rat renal mesangial cells. AB - 1. We have studied whether a nucleotide receptor mediates the effects of extracellular ATP and UTP on phosphatidylcholine metabolism in rat cultured glomerular mesangial cells. 2. ATP and UTP stimulated a biphasic 1,2 diacylglycerol (DAG) formation in [3H]-arachidonic acid-labelled mesangial cells. In contrast, in cells labelled with [3H]-myristic acid, a tracer that preferentially marks phosphatidylcholine, both nucleotides induced a delayed monophasic production of DAG with a concomitant increase in phosphatidic acid and choline formation. 3. A phospholipase D-mediated phosphatidylcholine hydrolysis was further suggested by the observation that ATP and UTP stimulate the accumulation of phosphatidylethanol, when ethanol was added to mesangial cells. 4. The rank order of potency of a series of nucleotide analogues for stimulation of phosphatidylethanol formation was UTP = ATP > ITP > ATP gamma S > beta gamma imido-ATP = ADP > 2-methylthio-ATP = beta gamma-methylene-ATP = ADP beta S, while AMP, adenosine, CTP and GTP were inactive, indicating the presence of a nucleotide receptor. 5. Elevation of cytosolic free Ca2+ by the calcium ionophore A23187 (1 microM) or the Ca(2+)-ATPase inhibitor, thapsigargin (200 nM) slightly increased phosphatidylethanol formation. However, chelation of cytosolic Ca2+ with high concentrations of Quin 2 did not attenuate ATP- and UTP-induced phosphatidylethanol production, thus suggesting that Ca2+ is not crucially involved in agonist-stimulated phospholipase D activation. 6. The protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA), but not the biologically inactive 4 alpha-phorbol 12,13-didecanoate, increased phospholipase D activity in mesangial cells, suggesting that PKC may mediate nucleotide-induced phosphatidylcholine hydrolysis. 7. Down-regulation of PKC-alpha and -delta isoenzymes by 8 h PMA treatment still resulted in full phospholipase D activation. In contrast, a 24 h treatment of mesangial cells with PMA, a regimen that also causes depletion of PKC-epsilon, markedly attenuated nucleotide-evoked phosphatidylethanol formation. In addition, the selective PKC inhibitor, calphostin C attenuated ATP- and UTP-induced phosphatidylethanol production.8. In summary, these data suggest that extracellular ATP and UTP use a common nucleotide receptor to activate phospholipase D-mediated phosphatidylcholine hydrolysis. Stimulation of phospholipase D appears to involve the PKC-epsilon isoenzyme, activated by DAG derived from phosphoinositide hydrolysis by phospholipase C. PMID- 8242261 TI - Characterization of the adenosine receptors of the rat superior cervical ganglion. AB - 1. Adenosine analogues caused hyperpolarization and inhibition of the depolarizing response to muscarine of the rat isolated superior cervical ganglion (SCG) measured by a 'grease gap' recording technique. The receptors mediating these responses have been characterized by use of a range of selective adenosine analogues and adenosine receptor antagonists. 2. In decreasing order of potency N6-cyclopentyladenosine (CPA), 2-chloroadenosine (2CA), adenosine, 2 phenylaminoadenosine (PAA), caused concentration-dependent hyperpolarizations whilst N6-(9-fluorenylmethyl)adenosine (PD 117,413) was inactive at up to 100 microM. 3. The order of potency of adenosine analogues in depressing depolarization caused by a submaximal concentration of muscarine (100 nM) was: CPA > R-PIA = 2CA > NECA > S-PIA > BZA > adenosine > PAA, where R- and S-PIA = R( )- and S(+)-N6-(2-phenylisopropyl)adenosine, NECA = 5'N-ethylcarboxamidoadenosine and BZA = N6-benzyladenosine. PD 117,413 was inactive at concentrations up to 100 microM. The maximum inhibitions of the muscarine-induced depolarization by CPA, 2CA, NECA and BZA were similar. R-PIA, S-PIA and PAA produced similar maximal inhibitions which were significantly smaller than those produced by CPA. 4. Hyperpolarizations caused by adenosine were antagonized by the P1-purinoceptor selective antagonist 1,3-dimethyl-8-phenylxanthine (8PT) and by the selective A1 adenosine receptor antagonist, 1,3-dipropyl-8-(4-((2 aminoethyl)amino)carbonylmethyloxyphenyl++ +)xanthine (XAC). Hyperpolarizations caused by CPA, adenosine and PAA were antagonized by the A1-selective antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) but not by the A2-selective antagonist, 3,7-dimethyl-1-propargylxanthine (DMPX). 5. Inhibition of the muscarinic-induced depolarization by CPA was antagonized by 8PT and DPCPXbut not by DMPX.6. It is concluded that the neurones of the rat SCG possess P1 purinoceptors of the Al-adenosine receptor subtype which mediate hyperpolarization and inhibition of depolarization caused by muscarine. PMID- 8242262 TI - Serum corticosterone, interleukin-1 and tumour necrosis factor in rat experimental endotoxaemia: comparison between Lewis and Wistar strains. AB - 1. Circulating corticosterone, interleukin-1 (IL-1) and tumour necrosis factor alpha (TNF alpha) activities in serum of Lewis and Wistar rats were measured following injection of lipopolysaccharide (LPS). IL-1 was measured as 'lymphocyte activation factor' (LAF) activity following precipitation of inhibitory activity with polyethylene glycol. TNF alpha activity was measured as cytotoxic activity. 2. Compared to the Wistar, the Lewis rat had higher circulating LAF and TNF activities following LPS, and release of both cytokines was prolonged in this strain. 3. Corticosterone increases in response to LPS were less in the Lewis than in the Wistar rat following the initial peak at 1 h; basal corticosterone was lower in the Lewis rat. 4. Adrenalectomized Lewis rats had even greater amounts of circulating LAF and TNF activities following LPS than did intact animals; the effect of adrenalectomy was not however mimicked by acute treatment with the steroid receptor antagonist, RU486, suggesting that endogenous corticosteroids did not acutely control cytokine release. 5. Although in vivo administration of anti-murine IL-1 alpha antiserum significantly lowered LAF activity of serum, circulating corticosterone in response to LPS was not affected. Similarly, treatment with anti-murine TNF alpha monoclonal antibody (mAb) abrogated TNF activity without affecting corticosterone, suggesting that other mediators may be responsible for corticosterone release following LPS. 6. This 'overproduction' of inflammatory cytokines together with lower circulating corticosterone may contribute to the susceptibility of the Lewis rat to diseases such as adjuvant arthritis or experimental allergic encephalomyelitis. PMID- 8242263 TI - Eicosanoid-induced Ca2+ release and sustained contraction in Ca(2+)-free media are mediated by different signal transduction pathways in rat aorta. AB - 1. The effects of 12-O-tetradecanoyl 4 beta-phorbol 13-acetate (beta-TPA) on the inositol 1,4,5-trisphosphate (IP3) production, Ca2+ release from the intracellular Ca2+ stores and sensitization of contractile apparatus, induced by prostaglandin F2 alpha (PGF2 alpha) and U46619, a thromboxane A2-mimetic, were studied, using fura-2-loaded and -unloaded rat thoracic aortic strips. 2. Both eicosanoids had characteristic patterns of responses in Ca(2+)-free, 2 mM EGTA containing solution (Ca(2+)-free solution). They induced transient increases in intracellular Ca2+ concentration ([Ca2+]i) without corresponding transient contraction, but produced delayed, sustained contraction, where [Ca2+]i was returned to the basal level. 3. Treatment with beta-TPA for 60 min reduced the eicosanoids-induced IP3 production, suggesting that the treatment inhibits PIP2 breakdown. 4. The treatment also attenuated [Ca2+]i transient induced by the eicosanoids, but not by caffeine (an IP3-independent releaser of stored Ca2+), in fura-2-loaded preparations incubated in Ca(2+)-free solution. 5. In contrast in the presence of beta-TPA, the sustained contractions evoked by the eicosanoids in Ca(2+)-free solution were potentiated, suggesting that the sites of actions of beta-TPA and the eicosanoids may differ from each other. 6. PGF2 alpha and U46619 utilize different and parallel signal transduction pathways to release Ca2+ by IP3 produced by PIP2 breakdown (beta-TPA-sensitive), and to increase the sensitivity of contractile apparatus, in which protein kinase C may not be involved (beta-TPA-insensitive). PMID- 8242264 TI - Role of oxygen radicals and arachidonic acid metabolites in the reverse passive Arthus reaction and carrageenin paw oedema in the rat. AB - 1. The role of arachidonic acid metabolites and oxygen radicals in carrageenin induced rat paw oedema and dermal reverse passive Arthus reaction (RPA) have been investigated. 2. Indomethacin (10 mg kg-1, p.o.) inhibited carrageenin paw oedema when administered 30 min before, but not 2 h after carrageenin. BWB70C (10 mg kg 1, p.o.), a selective inhibitor of 5-lipoxygenase, had no effect whether administered before or after carrageenin. Administration of both indomethacin and BWB70C had no greater anti-inflammatory effect than indomethacin alone. 3. BW755C (20 mg kg-1, p.o.), which inhibits the cyclo-oxygenase and lipoxygenase pathways of arachidonic acid metabolism, or superoxide dismutase-polyethylene glycol conjugate (SOD-PEG, 3000 u, i.v.) inhibited carrageenin paw oedema whether administered either 30 min before, or 2 h after carrageenin. 4. Pretreatment with dexamethasone (0.1 mg kg-1) or colchicine (2 mg kg-1), likewise suppressed carrageenin paw oedema. 5. BW755C (25-100 mg kg-1, p.o.) dose-dependently reduced plasma leakage in the RPA, whereas indomethacin (5 mg kg-1, p.o.) or BWB70C either alone or in combination, did not. 6. SOD-PEG (300-3000 u, i.v.) dose dependently inhibited plasma leakage in the RPA. In addition, the iron chelator and peroxyl radical scavenger, desferrioxamine (200 mg kg-1, s.c.) also inhibited plasma leakage. 7. Pretreatment with dexamethasone (0.1 mg kg-1) or colchicine (1 mg kg-1) reduced the plasma leakage in RPA, whereas MK-886 (10 mg kg-1) had no effect. 8. These results indicate an important role for oxygen radicals but not arachidonic acid metabolites in the maintenance of carrageenin paw oedema and the plasma leakage in RPA. Furthermore, the results suggest that the anti inflammatory actions of BW755C can be dissociated from its effects on arachidonic acid metabolism and are attributed to its anti-oxidant activity. PMID- 8242265 TI - Electrophysiological actions of alfentanil: intracellular studies in the rat locus coeruleus neurones. AB - 1. The electrophysiological effects of alfentanil on 156 neurones of the rat locus coeruleus were investigated by use of intracellular recordings from the in vitro brain slice preparation. 2. Bath application of alfentanil (5-100 nM) reversibly decreased the firing rate of all neurones tested in a dose-dependent manner, with an IC50 4.1.nM. 3. Based on inhibition of the spontaneous firing rate, alfentanil was 22 times more potent than morphine. 4. At 100 nM, alfentanil produced a complete inhibition of firing of all neurones tested (n = 62); the inhibition was accompanied by a membrane hyperpolarization 17.0 +/- 0.8 mV (range 6.1-30.3 mV, n = 62) and a reduction in input resistance 26.4 +/- 1.7% (range 6.5 53%, n = 51). 5. The effects of alfentanil were antagonized by naloxone, with a dissociation equilibrium constant of 2.7 +/- 0.4 nM (n = 6). 6. The reversal potential for the alfentanil-induced hyperpolarization was -110 +/- 2 mV (n = 9), which is approximately the potassium equilibrium potential. 7. The alfentanil induced hyperpolarization was blocked by caesium chloride and barium chloride. 8. These results indicate that alfentanil binds to mu-opioid receptors on the cell membrane of neurones of the locus coeruleus. This leads to opening of the inward going rectification potassium channels, resulting in the observed hyperpolarization of the membrane. PMID- 8242266 TI - Proceedings of The British Pharmacological Society Meeting. July 14-16, 1993. Abstracts. PMID- 8242267 TI - What do we expect from the International Federation of Sports Medicine (FIMS)? PMID- 8242268 TI - Gender verification--what purpose? What price? PMID- 8242269 TI - The XY female in sport: the controversy continues. AB - The summer and winter Olympic Games have been accompanied by much press coverage of the controversy and confusion over sex tests for sportswomen. Much of this has centred on the eligibility of subjects with androgen insensitivity to compete in women's events. The purpose of this paper is to review the process of sex differentiation and its abnormalities, highlighting those conditions in which biologically active testosterone is secreted which might confer an advantage in women's sporting events. PMID- 8242270 TI - A longitudinal study of exercise metabolism during recovery from viral illness. AB - An elite athlete engaged in a longitudinal programme of physiological assessment suffered a loss of performance that was later ascribed to an asymptomatic viral illness. In this 15-month, single subject, repeated measures study it was possible to demonstrate a severe decrease of exercise performance following viral illness. The oxygen uptake equivalent to the onset of blood lactate accumulation of 4 mmol l-1 (OBLA) decreased by 17% following viral illness, then recovered slowly, but had not returned to pre-viral levels 50 weeks later. Steady state exercise metabolism was also affected. During prolonged exercise at 70% of VO2max the proportion of energy derived from carbohydrate metabolism increased and an inability to maintain euglycaemia was observed. In both the graded and prolonged exercise tests, a pronounced tachycardia was evident after viral illness, possibly as a result of a febrile response to the infection, or, a direct effect of the virus upon myocardial performance. These data provide some preliminary and unique findings of the effects of viral illness upon the physiological and metabolic responses to exercise in an elite athlete. The data also demonstrate the sensitivity of currently used laboratory measures of exercise performance to monitor the changes in physiological function during recovery from viral illness. PMID- 8242271 TI - Physiological and psychological responses to a university fitness session. AB - The purpose of this study was to examine the physiological and psychological responses to a university fitness session entitled 'popmobility'. A popmobility session consists of 20 min of aerobic activities, 5 min of local muscular endurance exercises and 5 min of flexibility exercises. Ten regular participants of these sessions, women of mean(s.d.) age 21.2(1.5) years, took part in the study. A maximal oxygen uptake (VO2max) treadmill test was performed by each subject to obtain VO2max and maximum heart rate values. In a laboratory, heart rate and VO2 were measured throughout a popmobility session for each subject. Rate of perceived exertion (RPE) was measured every 5 min throughout the session. The mean intensity of the aerobic part of the session ranged from 67.7-82.6% of the subject's VO2max (mean of 76.4% VO2max). The mean heart rate reserve for the aerobic section was 75.6%. While the relative oxygen consumption remained fairly static during the aerobic section, the RPE score rose. The mean(s.d.) total energy expenditure was 236.6(28.4) kcal (range 203-288). The popmobility session is of adequate intensity to improve the aerobic fitness of its participants. Heart rate, as used as a measure of intensity during a popmobility session, would appear to be a fairly accurate indicator of intensity. However, the use of RPE for exercise prescription in popmobility sessions is inappropriate. Popmobility could also be useful in a weight-reduction programme. PMID- 8242272 TI - Re-examination of the incidence of exercise-induced hypoxaemia in highly trained subjects. AB - The purpose of this study was to examine the occurrence of exercise-induced hypoxaemia (EIH) during maximal exercise in highly trained athletes. Eleven trained cyclists (mean(s.d.) age 23(3.5) years; mean(s.d.) VO2max 66.9(4.8) ml kg 1min-1) performed a continuous, multistage (270 kpm min-1) cycle ergometer test to exhaustion. Measurements of arterial oxygen-haemoglobin saturation (%HbO2) were obtained simultaneously at rest, every 2 min during exercise, and at maximum exercise capacity from arterial blood sampling (%SaO2) and ear oximetry (%SpO2). Exercise induced hypoxaemia (%HbO2 < or = 91%) was present in 64% of the athletes examined when EIH was determined using pulse oximetry, whereas none of the subjects exhibited EIH when %HbO2 was determined using arterial blood. At rest the values for %HbO2 were similar with mean(s.d.) %SaO2 being 97.3(0.6)% and mean(s.d.) %SpO2 being 96.5(1.6)%. During exercise, statistically significant differences were found for %HbO2 between arterial blood and ear oximetry at the 6 min, 8-min, and maximal exercise sampling times (repeated measures analysis of variance, P < 0.05). The results indicate that ear oximetry overestimates the incidence of EIH and underestimates the oxyhaemoglobin saturation in highly trained cyclists during exercise in comparison with those measurements made from arterial blood. PMID- 8242273 TI - Seasonal variation of selected performance parameters in epee fencers. AB - In this study, anthropometric measurements were carried out on seven British international male epee fencers, using a maximal treadmill running test, a 20-s Wingate-type test, and isokinetic dynamometry. Testing was conducted on two occasions, 5 to 6 months apart, during mid-off-season (preparation) and mid-in season (competition) periods. Maximal oxygen intake (VO2max) and maximal respiratory exchange ratio (Rmax) were among the parameters obtained from the treadmill test, while peak and mean anaerobic power outputs were measured during a 20-s maximal effort. Knee extensor and flexor muscle forces from both dominant (leading) and non-dominant (trailing) legs were assessed at 1.04, 3.14 and 4.19 rad sec-1. Statistical analyses revealed lower mean VO2max (P < 0.05) and mean Rmax values (P < 0.02) at the in-season assessments compared with off-season. In season testing also demonstrated significantly lower peak torques for both dominant and non-dominant knee extensors compared with off-season assessments at all velocities (P < 0.05 to P < 0.004). Furthermore, in-season peak torque for the non-dominant leg flexors was lower (P < 0.03) at 4.19 rad sec-1 than off season. We conclude that current training practices may account for the observed seasonal variations in performance related physiological parameters in fencers. PMID- 8242274 TI - Regular exercise reduces fibrinogen levels: a review of longitudinal studies. AB - Fibrinogen can be considered as a major cardiovascular risk factor. Therefore there is a growing interest in methods to lower its plasma level. This paper is aimed at reviewing the data investigating the effects of regular exercise on plasma fibrinogen levels. Cross-sectional studies strongly suggest that regular exercise will reduce fibrinogen levels, and longitudinal data confirm this. The average decrease achieved by regular endurance exercise over several months is around 0.4 g/l. It can be extrapolated that this corresponds to a substantial decrease in risk of coronary heart disease. It is concluded that regular exercise is the most practicable approach known to date to lower plasma fibrinogen levels. PMID- 8242275 TI - High thoracic disc prolapse in a rugby player: the first reported case. AB - Traumatic disc prolapse in the thoracic region is not common, and is rare above the T3 level. A case of a T1-T2 disc prolapse as a result of rugby injury is described. PMID- 8242276 TI - Effect of continuous slow-speed running for 12 weeks on 10-14-year-old Indian boys. AB - Endurance training was conducted on a group of 41 East Indian boys aged 10-14 years and was compared with 25 untrained boys of the same age. A continuous slow running method was adopted for 12 weeks. The intensity of the training was 80-85% of maximum heart rate and frequency was 3 days per week. The boys were trained for a 1500-m event and therefore they covered three to five times their racing distance. For psychological reasons the training was carried out in a playground. The investigations included different physical and motor fitness tests: measurement of flexibility, agility, speed, leg muscle strength etc. Their performance times were also recorded before and after training. From statistical analysis we concluded that this particular type of training programme did not produce any detrimental effects on 10-14-year-old boys. On the other hand, this type of training did have some influence on improving physiological parameters in this age group of boys when compared with untrained boys of the same age. PMID- 8242277 TI - Influence of 6-week, 6 days per week, training on pituitary function in recreational athletes. AB - The influence on pituitary function of 6 weeks of training on 6 days a week was examined in six recreational athletes. Endurance training on a bicycle ergometer for 31-33 min was performed on 4 days each week at 90-96% (weeks 1-3) and 89-92% (weeks 4-6) of the 4 mmol lactate thresholds determined on day 0 and day 21, respectively, with interval training of 3-5 x 3-5 min in addition on 2 days a week at 117-127% and 115-110%, respectively. Determination of the serum hormone levels and a combined pituitary function test (200 micrograms thyrotropin releasing hormone (TRH), 100 micrograms gonadotrophin-releasing hormone (GnRH), 100 micrograms corticotrophin releasing hormone (CRH), 50 micrograms growth hormone releasing hormone (GHRH)) were made before training, after 6 weeks of training and after another 3 weeks of recovery. Training increased performance at 2 mmol lactate by 25%, at 4 mmol by 12%, and maximum performance by approximately 12%. The releasing hormone-stimulable prolactin, thyroid stimulating hormone (TSH) and somatotrophic hormone (STH) synthesis-secretion capacity remained unchanged, the adrenocorticotrophic hormone (ACTH) was increased after training. Cortisol release was reduced, follicle-stimulating hormone (FSH)-synthesis secretion capacity was increased after training, and the luteinizing hormone (LH) synthesis-secretion capacity reduced. This had no influence on base or exercise induced serum hormone levels (cortisol, aldosterone, insulin, prolactin, FSH, LH, TSH, ACTH, ADH and STH), which showed no dependence on training, except for free testosterone which showed a decreasing trend (P < 0.10) of 19-25% and post exercise ACTH which showed an increasing trend of 33% (P < 0.10). Conditioning (cortisol sensitivity and ACTH response) or adaptation (FSH and LH responses) to changed testosterone serum levels and altered spermatogenesis is discussed. PMID- 8242278 TI - Analysis of the work rates and heart rates of association football referees. AB - This study aimed to describe the work-rate profiles of referees during soccer matches and record heart-rate responses during these games. Using video recordings 14 referees were observed and their heart rates during the games were monitored by short-range radio telemetry. These included 11 football league matches. The exercise intensity was largely submaximal with a change in activity every 6 s. The mean distance covered during the game was 9.44 km; a significant fall in work rate was noted in the second half (P < 0.05). The mean heart rate of 165 beats min-1 did not vary between first and second halves. The work rate and heart rate varied more between individuals than with the importance of the match, but this variation was small. It was considered that refereeing top level soccer places high physiological demands on the official. This has implications for training and fitness assessment. PMID- 8242279 TI - Attitudes to mouthguards and prevalence of orofacial injuries in four teams competing at the second Rugby World Cup. AB - The opportunity was taken to conduct a questionnaire survey of the members of four competing countries at the second Rugby World Cup tournament (Australia, Ireland, Scotland, Wales) regarding the prevalence of orofacial injuries and attitudes to mouthguards. The results show that while all players in each team believed that mouthguards provided local protection, approximately one in five of each team did not wear one. The average age when players first started wearing mouthguards varied from 12.7 years for Australia to 18.1 years for Wales, while the percentage of wearers willing to play without their mouthguard ranged from only 4.6% for Australia to 15.8% for Wales. Just less than half of the total group had previously sustained an orofacial injury playing rugby. PMID- 8242280 TI - Anabolic steroid use among students at a British college of technology. AB - To determine the rate of current or previous use of anabolic steroids by students at a UK college of technology, a questionnaire survey of 687 day students was conducted. The questionnaire began with a general section for all of the students, which ended with the question 'Have you ever used anabolic steroids?'. A further section specifically for anabolic steroid users examined patterns of use, and how certain circumstances might affect the individual's decision to use anabolic steroids. The response rate to the questionnaire was 92%. The overall rate of current or previous use of anabolic steroids was 2.8% (4.4% in males, 1.0% in females). Of these, 56% had first used anabolic steroids at age 15 or less. Anabolic steroid users were more likely to be male, under 17 years of age, and participating in bodybuilding, weight-lifting or rugby. The results of this survey, if confirmed in other groups of young people, would suggest that use of anabolic steroids may be widespread in the UK. PMID- 8242281 TI - Validity of 20-MST for predicting VO2max of adult Singaporean athletes. AB - This investigation compared the results of direct and indirect measurements of maximum oxygen uptake (VO2max) of an Asian population. Twenty subjects (16 male, 4 female), all of whom were physical education students, were assessed directly using laboratory treadmill running to determine VO2max. The indirect estimation of VO2max was obtained using a modified form of the Leger and Lambert 20-m multistage shuttle run test (20-MST). Heart rates were recorded throughout both tests. Pearson product moment correlations confirmed test-retest reliability for both direct and indirect measurements (r = 0.90 and r = 0.91 respectively). Differences for test-retest were found to be not significant. No differences were found between the maximal heart rate responses of the subjects for the direct and indirect tests. Of the subjects 75% had a lower predicted VO2max value (P < 0.01) compared with results gained by direct measurements when the Ramsbottom norms for the 20-MST were used. The reasons for this difference could be due to the different racial groups used as subjects, the climatic conditions in Singapore, or the small sample size. In order for the 20-MST test to be considered a valid measure of aerobic fitness in Singapore with an Asian population further study is recommended. PMID- 8242282 TI - Effect of intravenous fluid administration on recovery after running a marathon. AB - After the Rotterdam Marathon on 21 April 1991 (ambient temperature 5.8 degrees C, relative humidity 74%, wind velocity 5 m s-1) data from 66 athletes were analysed for information concerning total recovery and recovery from pain, stiffness, loss of appetite, sleep disturbance and fatigue. The pulse rate, body weight and temperature were measured. The athletes were divided at random into two groups. Thirty-four athletes received an intravenous infusion of 2.5 l of a 2.5% glucose/0.45% NaCl solution. Thirty-two athletes received a placebo infusion of 100 ml 0.9% NaCl. Recovery took 9.2 days in the placebo group and 10.2 days in the infusion group. All athletes had pain and/or stiffness after the marathon. The immediate replacement of 2.5 l of fluid had no significant influence on the rate of total recovery, the number of days with pain or stiffness, the appetite, sleep or fatigue. On the first day after the marathon the pulse rate was increased. The rectal temperature was not affected. The athletes were also divided into fast and slow runners without regard to fluid replacement. Fast runners (those running the race in less than 2 h 55 min) needed more time to recover than slower runners and pain and/or stiffness lasted longer in the fast group. Athletes who equalled or improved their best previous results also needed more time to recover than athletes who did not, although there were no significant differences in pain and stiffness. Athletes did not benefit from immediate fluid replacement after running the 1991 Rotterdam Marathon. PMID- 8242283 TI - Trauma centres. PMID- 8242284 TI - Laparoscopic surgery for colorectal cancer. PMID- 8242285 TI - Complications of circumcision. AB - Circumcision remains a common operation, with over 30,000 procedures performed annually in the UK, mostly on children. The British Medical Association has recommended that circumcision should be performed only for medical reasons. Despite this, controversy exists over whether too many circumcisions are being performed. Are patients being exposed to an unnecessary operation? It may be argued that in doubtful cases it is easier to proceed to circumcision on the assumption that the attendant risks are low, but the operation is associated with a definite morbidity and rare deaths have been reported. This review considers the spectrum of complications that may result from circumcision and discusses the possible aetiological mechanisms. PMID- 8242286 TI - Infrainguinal revascularization in the diabetic patient. AB - This article reviews the epidemiological, anatomical and physiological background to infrainguinal arterial reconstruction in the diabetic patient. There is no firm evidence in the literature for the existence of a surgically significant occlusive 'small-vessel' disease in diabetic patients, either as an aetiological component in the pathogenesis of diabetic foot lesions or as a factor limiting the success of infrainguinal bypass grafting. Recent published results of infrainguinal vascular reconstruction in patients with diabetes are reviewed. These are encouraging; early aggressive treatment of lower-limb occlusive disease may be warranted in the diabetic patient. PMID- 8242287 TI - Smoking impairs endothelium-dependent relaxation of saphenous vein. AB - Samples of proximal saphenous vein were obtained from heavy smokers and non smokers: 28 were obtained at varicose vein surgery and eight at infrainguinal bypass surgery. The veins were prepared for histological examination and for mounting in an organ bath to measure changes in isometric tension. Vein rings from both smokers and non-smokers responded equally to sodium nitroprusside, a reagent that acts directly on smooth muscle cells, undergoing 90-100 per cent relaxation. The mean(s.e.m.) maximum relaxation in response to bradykinin of rings obtained at varicose vein surgery was 50.0(5.3) per cent in non-smokers compared with only 31.6(2.2) per cent in smokers (P = 0.03). Similarly, in specimens obtained at bypass surgery the mean(s.e.m.) maximum relaxation in smokers was only 25.1(6.3) per cent compared with 48.4(4.1) per cent in smokers (P = 0.04). Relaxation in response to the bradykinin stimulus was abolished in the presence of L-nitroarginine methyl ester, a specific inhibitor of the synthesis of endothelium-derived relaxing factor (EDRF). Preincubation of the vein rings with L-arginine, the precursor of EDRF, did not increase the vasorelaxation in smokers. The mean(s.e.m.) maximum relaxation in response to the calcium ionophore A23187 was 53.5(3.8) per cent in non-smokers compared with only 27.0(4.9) per cent in smokers (P = 0.01). The results indicate that heavy smokers have impaired release of EDRF in response to both bradykinin and calcium ionophore. This impairment may increase vasomotor tone, platelet aggregation and smooth muscle proliferation, thereby resulting in an increased risk of graft occlusion. PMID- 8242288 TI - Influence of type III collagen genotype on aortic diameter and disease. AB - Type III collagen contributes to the tensile strength of the aortic wall, and mutations in the type III collagen gene have been suggested as the basis for the familial tendency to abdominal aortic aneurysm (AAA). Variation in this gene was investigated in 153 patients with AAA, 87 with aortoiliac stenosis and 98 age matched controls. The rare mutation at amino acid 619 of Gly-->Arg, previously associated with AAA in a single family, was not found in any of the patients with aneurysm. For the Ala-->Thr variation at amino acid 531, the frequency of the threonine allele was 0.25 in patients with AAA and stenosis, compared with 0.35 in controls. The frequency of the rare allele in the region 3' to the gene demonstrated by Ava II digestion (0.27 in the general population) was found to be 0.29 in the AAA group and 0.19 in the stenosis group (P = 0.023). In the AAA group the presence of the Ava II rare allele was associated with a significant increase in aneurysm diameter (P = 0.044). Non-invasive assessment of aortic distensibility was available in 25 patients: those carrying the Ava II rare allele had less distensible aortas than those not carrying this allele (median pressure-strain elastic modulus 42.0 and 23.9 N/cm2 respectively, P = 0.008). Variation in the type III collagen gene may influence the mechanical properties of the ageing aorta and hence its susceptibility to disease and dilatation. In contrast, there is no evidence for a single common founder mutation in type III collagen predisposing to AAA. PMID- 8242289 TI - Early detection of occult colorectal hepatic metastases using duplex colour Doppler sonography. AB - Previous studies have shown that overt intrahepatic tumours are associated with subtle changes in liver perfusion that can be measured directly using duplex colour Doppler sonography (DCDS). This study assessed the predictive value of DCDS in the early detection of occult colorectal hepatic metastases. Hepatic arterial and portal venous blood flow was measured in 50 control subjects and 135 patients with colorectal cancer, 67 with overt liver metastases and 68 with an apparently disease-free liver on the basis of computed tomography and laparotomy findings. The Doppler perfusion index (DPI), defined as the ratio of hepatic arterial to total liver blood flow, was calculated. Clear separation of the DPI values of controls and those of patients with overt metastases was observed (P < 0.0001). Thirty-eight of the 68 patients with a disease-free liver also had an abnormally high DPI value. After 1 year of follow-up, 21 patients with an abnormally high DPI at the time of apparently curative primary resection had developed liver metastases and a further four had died without post-mortem examination. The 30 patients with normal DPI remain disease-free. The data suggest that DPI is of value in the early detection of occult liver metastases. PMID- 8242290 TI - Ventilatory and blood gas changes during laparoscopic and open cholecystectomy. AB - As part of a randomized trial, ventilatory and arterial blood gas changes were assessed during open (n = 30) and laparoscopic (n = 30) cholecystectomy. Measurements were made during anaesthesia before the start of surgery and at the time of removal of the gallbladder. Despite an increase in minute ventilation from a mean(s.d.) of 5.7(1.4) to 6.1(1.2) litres, mean(s.d.) arterial carbon dioxide tension (PaCO2) rose from 5.3(0.9) to 6.0(0.9) kPa during laparoscopic cholecystectomy. End-tidal carbon dioxide tension (PE'CO2) had poor precision in predicting PaCO2 (95 per cent interval of agreement -0.61 to 1.93 kPa). Mean(s.d.) peak airway pressure increased from 17(4) to 23(4) cmH2O. The mean PaCO2--PE'CO2 value did not change significantly, although there was significant within-patient variation. Arterial oxygen levels did not change significantly. By comparison, no clinically significant changes in ventilation or blood gas values occurred during open cholecystectomy. In conclusion, laparoscopic cholecystectomy requires a substantial but variable increase in minute ventilation to compensate for carbon dioxide absorption from the peritoneum. PMID- 8242291 TI - Comparison of metabolic responses to laparoscopic and minilaparotomy cholecystectomy. AB - This randomized study compared the metabolic responses to laparoscopic cholecystectomy (n = 10) and minilaparotomy cholecystectomy with a 5-7-cm incision (n = 10). Venous blood samples were taken before operation and at 3, 6, 9, 12, 18, 24, 48, 72 and 168 h after incision and analysed for levels of C reactive protein, interleukin 6, cortisol, albumin, transferrin, iron, fibrinogen, fibrin degradation products and polymorphonuclear elastase, and for neutrophil and lymphocyte counts. Urine samples (24 h) were analysed for urea, creatinine, 3-methylhistidine and catecholamines. The magnitude of the metabolic changes from baseline levels was quantified by calculating areas under each individual curve. A significant metabolic response with a similar time course and magnitude of changes occurred after laparoscopic and minilaparotomy cholecystectomy but with wide variation in magnitude between individuals. PMID- 8242292 TI - Multidrug-resistant colonic cancer cell line LoVoDx is efficiently killed by lymphokine-activated killer cells from patients with carcinoma of the colon. AB - Multidrug-resistant (MDR+) cancer cells have the ability to grow in the presence of cytotoxic concentrations of antineoplastic drugs as a result of possessing the transmembrane drug efflux pump p-glycoprotein. The MDR+ colonic cancer cell line LoVoDx (derived from the drug-sensitive line LoVo) was tested for sensitivity to lymphokine-activated killer (LAK) cell-mediated toxicity. LAK cells were cultured from patients with colonic cancer and from matched controls with benign disorders. LAK cells from patients with cancer were as effective as those from controls in mediating cytotoxicity. The MDR+ cell line was significantly more sensitive to LAK cell-mediated cell killing than its parental drug-sensitive line LoVo (P < 0.05). These results indicate a possible role for adoptive immunotherapy in MDR+ tumours expressing p-glycoprotein. PMID- 8242293 TI - Long-term results of a single protocol for thrombolysis in acute lower-limb ischaemia. AB - In a prospective study, 78 of 157 patients with acute lower-limb ischaemia were considered suitable for thrombolysis. The immediate and 4-year results of 52 patients managed by a single protocol are reported. Of 34 patients alive at 4 years, 23 had limb salvage. Initial treatment produced effective lysis in 38 patients (73 per cent) with significant benefit in 35 (67 per cent); that benefit was sustained for a minimum of 4 years in 30 patients (58 per cent). Seven of the 18 deaths by 4 years occurred within 30 days. Amputation was carried out in six patients within 30 days and in five during the next 4 years. Delayed amputation followed persistent distal occlusion or progression of distal disease. No death or amputation was caused by complication of treatment. In selected patients the risks of thrombolysis can be reduced to an acceptable level by personal supervision and a strict protocol. In survivors, limb salvage is generally sustained for at least 4 years. PMID- 8242294 TI - Low-dose aspirin fails to inhibit increased platelet reactivity in patients with peripheral vascular disease. AB - A study was performed to evaluate the effect of low-dose aspirin (75 mg/day) on platelet reactivity in patients with peripheral vascular disease. Platelet function was measured in 31 patients with advanced peripheral vascular disease requiring surgery. Shear-induced haemostasis and collagen-induced thrombus formation were used as indicators of platelet reactivity. They were measured in non-anticoagulated fresh whole blood samples ex vivo using a haemostatometer. Results were compared with those from 30 age- and sex-matched controls. Shear induced haemostasis and collagen-induced thrombus formation were also measured in 18 of the 31 patients before the start of aspirin administration (before operation) and 1 week after surgery. Patients with advanced peripheral vascular disease were found to have increased platelet reactivity with a mean(s.e.m.) shear-induced haemostasis index of 1977(180) mmHg s compared with a control value of 3161(234) mmHg s (P < 0.001) and a mean(s.e.m.) collagen-induced thrombus formation index of 3980(460) mmHg s compared with a control value of 5350(420) mmHg s (P = 0.02). Perioperative low-dose aspirin failed to inhibit platelet function in patients with peripheral vascular disease. PMID- 8242295 TI - Changing trends in the management of lower-limb ischaemia: a 17-year review. AB - Over a 17-year period, 2930 vascular procedures were performed for chronic lower limb occlusive disease. During this time, the total vascular surgical workload rose significantly as a result of an increase in the rates of arterial reconstruction and percutaneous transluminal angioplasty. Although the major lower-limb amputation rate did not change significantly, there was a decrease in the above- to below-knee ratio. In addition, there were significant increases in the percentage of patients treated over the age of 75 years and the proportion of attempted revascularizations before amputation. The mortality rates for arterial reconstruction, percutaneous transluminal angioplasty and amputation did not change significantly during the study period. The duration of hospital stay increased significantly for patients undergoing amputation and decreased significantly for those receiving arterial reconstruction. PMID- 8242296 TI - Characterization of symptomatic and asymptomatic carotid plaques using high resolution real-time ultrasonography. AB - High-resolution ultrasonography was used to classify carotid plaques into five different types in 72 patients with symptoms and in 49 without, and with stenosis of the origin of the internal carotid artery > 70 per cent. There were 72 plaques in the symptomatic group and 75 in the asymptomatic group. Type 1 plaques were uniformly echolucent, type 2 predominantly echolucent, type 3 predominantly echogenic, type 4 uniformly echogenic and type 5 consisted of plaques that could not be classified owing to heavy calcification and acoustic shadows. Type 1 plaque was found in 90 per cent of patients with symptoms and in 10 per cent of those without, type 2 plaque was found in 53 and 47 per cent, type 3 in 34 and 66 per cent, and type 4 in 5 and 95 per cent, respectively. The preponderance of echolucent plaques in symptomatic patients with stenosis > 70 per cent supports the hypothesis that this type of plaque is unstable and tends to embolize. In contrast, in patients without symptoms there is preponderance of echogenic plaques. PMID- 8242297 TI - Serial imaging of the carotid bifurcation and cerebrovascular reserve after carotid endarterectomy. AB - A radioisotopic method of quantifying mean cerebral transit time was used to assess the immediate effects of carotid endarterectomy on cerebrovascular reserve (CVR) in 69 patients. In addition, serial postoperative data were acquired on CVR, clinical status and non-invasive imaging of the internal carotid arteries in 56 patients over a period of 6-48 (median 24) months. Twenty-one patients (30 per cent) had preoperative evidence of impaired CVR in the symptomatic hemisphere. Within 4 days of surgery, however, reserve had returned to normal in 17 of the 21 patients. During follow-up, four of the 56 patients developed recurrent stenosis (> 50 per cent) or occlusion of the artery operated on but only two of these had impairment of CVR and none was symptomatic. Three patients suffered recurrent transient ischaemic attacks (TIAs) but none had recurrent internal carotid artery disease or impaired CVR. One patient suffered a TIA in the territory of the non operated artery during follow-up in association with disease progression and CVR impairment. However, the TIA preceded recognition of either of these changes. Twelve other patients had (or developed) stenosis (> 50 per cent) in the non operated artery during follow-up but none was symptomatic or developed impairment of CVR. Although assessment of CVR provided useful information on the frequency of haemodynamic compromise before carotid endarterectomy and on the natural history of disease progression, neither serial assessment of reserve nor non invasive imaging of the carotid bifurcation influenced clinical practice during follow-up. PMID- 8242298 TI - Cryostripping the long and short saphenous veins. PMID- 8242299 TI - Suture length to wound length ratio and healing of midline laparotomy incisions. AB - The effect of suture length to wound length ratio on the healing of midline laparotomy wounds closed with a continuous suture was evaluated in a prospective clinical trial. All patients undergoing abdominal procedures through a midline incision were included except those with an incisional hernia after previous midline operation. The total incidence of wound infection was 36 of 454 patients (7.9 per cent) and wound dehiscence requiring reoperation occurred in three patients (0.7 per cent). Incisional hernia was found in 18.7 per cent of 363 patients alive 12 months after surgery. Multivariate analysis identified the suture length to wound length ratio, age and major wound infection as independent risk factors for the development of hernia, which occurred in 9.0 per cent of patients when the suture length to wound length ratio was > or = 4 and in 23.7 per cent (P = 0.001) when it was < 4. The suture length to wound length ratio is an important parameter for healing of midline incisions closed with a continuous suture technique. The incidence of incisional hernia is lower when such wounds are sutured with a ratio > or = 4. PMID- 8242300 TI - Complications and survival of multilumen central venous catheters used for total parenteral nutrition. AB - In a prospective study of 879 triple-lumen catheters, 219 pulmonary artery catheters, 31 double-lumen and six single-lumen catheters used for the administration of total parenteral nutrition over a 1-year period, the overall complication rate was 12.5 per cent (14.7 complications per 1000 catheter-days) and the catheter-related sepsis rate 4.4 per cent (5.2 per 1000 catheter-days). The probability of development of catheter-related sepsis did not increase with the duration of catheterization. There were no differences in the rate of complications associated with 427 catheters changed by replacement at a new site compared with 159 lines changed over a guidewire. These data support the use of multilumen central venous catheters for the administration of total parenteral nutrition. They suggest that a routine weekly change of line is unnecessary; catheters should be changed only on the development of a complication. When it is required, a catheter should be changed by replacement over a guidewire. PMID- 8242301 TI - On-scene medical decision making and overtriage. AB - This study examined whether a hospital-based helicopter trauma service led to 'overtriage' of patients back to the multispecialty base hospital. All accident site triage decisions made during 1991 were studied. Seven of the 574 decisions (1.2 per cent) led to inappropriate movement of the patient to a specialist centre. Twenty-six decisions (4.5 per cent) were based on mechanism of injury alone and the possibility of overtriage by this method is discussed. A trained doctor can select the correct destination for a patient with trauma without inappropriately using the base hospital in preference to local centres. PMID- 8242302 TI - Acute haemodilution in surgery for burns: a preliminary report. AB - Blood loss is a major problem in the management of patients with burns treated by early excision and grafting. Acute normovolaemic haemodilution has been successfully used in cardiac surgery for many years but has not been reported previously in surgery for burns. The technique has been used in six patients with burns undergoing early excision and grafting, with no obvious morbidity and an apparent reduction in homologous blood requirements. PMID- 8242303 TI - Early wound excision and skin grafting restores cellular immunity after severe burn trauma. AB - Suppression of cellular immunity following thermal injury may contribute to the high incidence of postburn sepsis. Early excision and skin grafting may remove deleterious local wound factors and prevent immunosuppression. The effect on cellular immunity of early burn wound excision and skin grafting was investigated in an animal model using female Wistar rats. The development of immunosuppression shortly after burn injury was shown by popliteal lymph node assay and a 2,4 dinitrofluorobenzene skin reaction test. Excision and skin grafting 2 days after burn trauma restored parameters to normal. Burned tissue suppresses cellular immunity; this effect is preventable by early excision and skin grafting. PMID- 8242304 TI - Axillary nodal metastases from an occult primary consistent with breast carcinoma. AB - The records of 15 women presenting with axillary nodal metastases from an occult primary consistent with breast carcinoma were reviewed. Nine patients underwent complete axillary clearance and in six the clinical mass only was excised. Radiotherapy to the axilla was given to eight patients. One woman underwent mastectomy at the time of axillary surgery but no tumour was found in the excised breast; in the remaining 14 patients the breast was left untreated. One premenopausal woman received adjuvant systemic chemotherapy and one postmenopausal patient was given tamoxifen. Three patients died from recurrent disease at 16, 50 and 56 months after treatment of the axillary mass; one is alive at 42 months with systemic recurrence. In two of these patients the tumour in the breast became apparent before other metastases. Eleven patients survived for a median of 92 (range 18-144) months without evidence of disease. The prognosis of this patient group is better than that generally reported for stage II breast carcinoma. PMID- 8242305 TI - Concurrent hyperthyroidism and thyroid carcinoma. AB - Between 1986 and 1991, thyroidectomy was performed on 138 patients with hyperthyroidism. Thyroid carcinoma was found in eight patients (5.8 per cent). Eighty (58.0 per cent) of the 138 patients had toxic nodular goitre, 33 (23.9 per cent) toxic diffuse goitre and 25 (18.1 per cent) toxic adenoma. Concurrent carcinoma was more frequent in patients with toxic adenoma (8 per cent) than in those with Graves' disease (6 per cent) and toxic nodular goitre (5 per cent). Papillary carcinoma was found in seven patients and follicular carcinoma in one. Three papillary carcinomas were occult with a diameter less than 1.5 cm. Five patients received 100 mCi 131I after operation. There was no morbidity. During follow-up of 10-45 months, there was neither death nor recurrence. PMID- 8242306 TI - Thyroid neoplasia coexistent with chronic lymphocytic thyroiditis. AB - During a 6-year period 115 patients presenting with thyroid enlargement had evidence of chronic lymphocytic thyroiditis on fine-needle aspiration cytology. Of 27 patients in whom histological analysis was carried out, 16 had neoplasms (follicular adenoma, four; follicular carcinoma, one; papillary carcinoma, four; lymphoma, seven). Assuming that neoplasia was not overlooked in the absence of histological examination, the overall incidence was 14 per cent and that of malignant disease 10 per cent. All patients with carcinoma had cytological features suspicious of neoplasia on the first or subsequent aspirates in addition to those of thyroiditis. Cytology was suspicious of lymphoma in only two of seven patients but increasing size of the thyroid swelling was a consistent feature. Evidence of chronic lymphocytic thyroiditis may lead to neoplasia being overlooked. Repeated cytological analysis is helpful in identifying coexistent carcinoma but unreliable in excluding lymphoma. PMID- 8242307 TI - Oesophagogastrectomy for carcinoma of the oesophagus and cardia. AB - A retrospective review was undertaken of all oesophagectomies performed within a single unit over a 12-year period. In all, 298 patients with primary oesophageal cancer underwent resection between March 1979 and December 1991. Four patients had a three-stage oesophagogastrectomy, 27 a thoracoabdominal oesophagogastrectomy and 267 a Lewis procedure. Dysphagia was the predominant presenting symptom. The duration of symptoms was not related to the stage of disease. Before diagnosis, 52 per cent of patients tolerated symptoms for 2-4 months. Adenocarcinoma was found in 180 tumours and squamous cell carcinoma in 103. Half of the patients had evidence of metastatic spread at the time of laparotomy or thoracotomy. The 30-day mortality rate was 10 per cent and the overall actuarial 5-year survival rate of all patients 23 per cent. The actuarial 5-year survival rate of patients without lymph node involvement was 39 per cent compared with 17 per cent for those with positive nodes (P < 0.05). Five of eight patients who had anastomotic leakage died. The almost unselected nature of this series, coupled with the favourable results of oesophagectomy, support the contention that resection remains the preferred mode of treatment for carcinoma of the oesophagus of all histological types. PMID- 8242308 TI - Non-operative treatment of small bowel obstruction following appendicectomy or operation on the ovary or tube. AB - This study examined the hypothesis that small bowel obstruction in patients whose only previous laparotomy was for appendicectomy or operation on the ovary or tube is unlikely to resolve with non-operative management. Results of 330 admissions for small bowel obstruction were examined. In 40 cases the only previous laparotomy was at appendicectomy or tubo-ovarian operation; in 38 (95 per cent) of these division of adhesions was undertaken compared with 154 (53 per cent) of the remaining 290 (P < 0.00001). In the former group band adhesions were commoner (86 versus 45 per cent, P < 0.00001), bowel resection was required more frequently (22 versus 10 per cent, P = 0.02) and it was considered safe to give a trial of non-operative management less often (60 versus 85 per cent, P = 0.0004). A trial of conservative management may be unsafe or not worth while in patients with obstruction following earlier appendicectomy or operation on the ovary or tube. PMID- 8242309 TI - Pancreatic fistula cured by an endoprosthesis in the pancreatic duct. PMID- 8242310 TI - Effect of nifedipine on renal transplant rejection. AB - The effect of early nifedipine therapy on acute renal allograft rejection was studied in 170 adult cadaveric transplant recipients. Acute rejection occurring in the first 3 months after transplantation was diagnosed by Tru-cut biopsy and the severity of each rejection episode assessed histologically. The incidence of acute rejection was significantly lower in patients treated with nifedipine (29 of 80; 36 per cent) than in controls (52 of 90; 58 per cent) (P < 0.01) and there was a higher proportion of histologically mild rejection episodes in the former group (P < 0.01). Multivariate analysis confirmed that nifedipine exerted a significant independent effect on the incidence of early acute rejection. Other factors identified in the multivariate model as influencing rejection were human leucocyte antigen (HLA) matching at the DR locus, blood level of cyclosporin during the first week, HLA matching at the B locus, donor age and donor sex. The 1-year graft survival rate was 88.6 per cent in patients given nifedipine and 63.8 per cent in controls (P < 0.02). These data suggest that nifedipine therapy has a useful role in human renal transplantation. PMID- 8242311 TI - Cytokine gene transcription in renal cell carcinoma. AB - Cytokines are powerful modulators of immune responses, the local production of which could be relevant to the interaction between tumour and immune system. This study investigated the transcription of genes encoding interleukin (IL) 2, IL-4, IL-10 and interferon (IFN) gamma in lymphocyte-infiltrated renal cell carcinoma biopsies from ten patients using the reverse polymerase chain reaction technique. Autologous peripheral blood mononuclear cells and healthy renal parenchyma tissue were tested in parallel. The beta-actin gene, used as a positive control, was transcribed in all samples. In contrast, transcription of cytokine genes was confined to tumour biopsies: IL-2 gene transcripts were detectable in five biopsies and IL-10 transcripts in seven. IL-4 and IFN-gamma gene transcripts were detectable in one biopsy each. In two patients no cytokine gene transcription could be identified. These data underline that heterogeneous patterns of cytokine gene transcription can be observed in renal cell carcinoma biopsies. Although transcription of an immunostimulatory lymphokine such as IL-2 was observed in 50 per cent of biopsies, the most frequently transcribed cytokine gene coded for an inhibitory factor, IL-10. PMID- 8242312 TI - Localized argyria: a complication of the repair of inguinal hernia with silver filigree. PMID- 8242313 TI - Faecal occult blood screening for patients with gastrointestinal symptoms. PMID- 8242314 TI - Delay in the diagnosis and prognosis of carcinoma of the right colon. AB - Of 152 consecutive patients with carcinoma of the right colon, 61 (40 per cent) suffered delays in treatment of more than 12 weeks from the onset of symptoms, with a mean delay of 48 weeks. The most common error was failure to initiate or complete the investigation of iron-deficiency anaemia (33 patients). False negative barium enema investigations occurred in 16 cases. Patients with delays in diagnosis had survival rates not significantly different from those who presented early. Thirty-one patients with anaemia and no abdominal symptoms had a significantly higher survival rate than 30 presenting with abdominal symptoms, despite delays in treatment (P < 0.02). Greater vigilance is required in the investigation of patients presenting with iron-deficiency anaemia. PMID- 8242315 TI - Colonoscopic surveillance after curative resection for colorectal cancer. AB - Colonoscopic surveillance after resection for colorectal cancer has been advocated to improve detection of anastomotic recurrence, and of synchronous and metachronous tumours. The benefit provided by colonoscopy remains unproven, and the best timing of examination is unclear. To determine the value of colonoscopy after curative resection for large bowel cancer, the efficacy of an endoscopic surveillance programme in the early detection of intraluminal bowel recurrence in a series of patients admitted with colorectal cancer was examined. Between April 1983 and December 1988, 132 patients underwent colonoscopy. Eight (6.1 per cent) were found to have intraluminal recurrence without evidence of extraluminal spread. Six of these recurrences were at the site of anastomosis and two represented metachronous tumour development. All of these patients were symptomatic at the time of diagnosis. In 15 patients (11.4 per cent), adenomatous polyps were discovered during the initial endoscopic examination. These results indicate that colonoscopic surveillance will rarely allow early detection of asymptomatic intraluminal bowel recurrence, but is valuable in the detection of synchronous lesions. Frequent surveillance is not justified in the early postoperative years and colonoscopy should probably be confined to a single procedure to exclude synchronous lesions. PMID- 8242316 TI - Visualization of the ureter during laparoscopic colonic resection. PMID- 8242317 TI - Initial report from a Swedish multicentre study examining the role of preoperative irradiation in the treatment of patients with resectable rectal carcinoma. Swedish Rectal Cancer Trial. AB - Between March 1987 and February 1990, 1168 patients with resectable rectal cancer were recruited into a nationwide Swedish trial. Patients were randomly allocated to receive either preoperative irradiation (25 Gy in five fractions over 1 week) followed by surgery within 1 week, or surgery alone. In each group 454 patients underwent curative surgery. The groups were well matched for age, sex and surgical procedure. The postoperative mortality rates did not differ: 4 per cent after irradiation versus 3 per cent after surgery alone, but the postoperative mortality rate was significantly higher in patients treated with a two- rather than three- or four-portal technique (15 versus 3 per cent, P < 0.001). More perineal wound infections were found in patients receiving irradiation (20 versus 10 per cent, P < 0.001). No difference in the incidence of anastomotic dehiscence or other postoperative complication was found between the groups. Preoperative radiotherapy with a short-term high-dose technique can be delivered without major acute adverse effects, provided the irradiation technique is optimized. PMID- 8242318 TI - In vitro response of the human anal canal longitudinal muscle layer to cholinergic and adrenergic stimulation: evidence of sphincter specialization. AB - A study was performed to determine the in vitro response of the longitudinal smooth muscle layer (conjoined longitudinal coat) of the human and canal to cholinergic and adrenergic stimulation, and to compare this with similar features of the internal anal sphincter (IAS) and rectal smooth muscle. Tissue was obtained from abdominoperineal and anterior resection specimens, and strips of muscle mounted for isometric tension recording in superfusion organ baths. Both conjoined longitudinal coat and IAS strips generated and maintained spontaneous myogenic tension (mean(s.e.m.) 0.10(0.01) and 0.48(0.04) g per mg tissue respectively), whereas equivalent rectal smooth muscle did not. Stimulation of muscarinic (cholinergic) receptors caused contraction of strips from the conjoined longitudinal coat and rectal smooth muscle layers in a dose-dependent manner (10(-7)-10(-4) mol/l); however, IAS strips relaxed (10(-6)-10(-4) mol/l). Activation of alpha-adrenergic receptors in conjoined longitudinal coat and IAS strips produced concentration-dependent contraction (10(-7)-10(-4) mol/l). In comparison, rectal smooth muscle relaxed. All muscle layers relaxed in response to beta-adrenoceptor stimulation. These data indicate that in the anal canal both the conjoined longitudinal coat and IAS are specialized sphincteric smooth muscles. PMID- 8242319 TI - Physiological and histochemical adaptation of the electrically stimulated gracilis muscle to neoanal sphincter function. AB - The physiological and histochemical characteristics of the gracilis muscle were studied in 19 patients undergoing electrically stimulated gracilis neosphincter construction. Indications for surgery were faecal incontinence (n = 11) and reconstruction following sphincter excision or congenital absence (n = 8). Transposition of the gracilis muscle around the anal canal followed by chronic low-frequency electrical stimulation was associated with a shift in the frequency response curve and a prolongation of the time-course of individual muscle twitches suggestive of transformation to a slow-twitch fatigue-resistant type. Temporary cessation of electrical stimulation resulted in a reversal of the frequency-response changes. Muscle biopsies taken before and a median of 80 (range 49-137) days after transposition and low-frequency electrical stimulation indicated a significant increase in the proportion of type 1 fibres and a significant decrease in their diameter. These results show that the human gracilis muscle is capable of physiological and histochemical adaptation to long term neosphincter function. PMID- 8242320 TI - Herniation at the site of cannula insertion after laparoscopic cholecystectomy. PMID- 8242321 TI - Herniation at the site of cannula insertion after laparoscopic cholecystectomy. PMID- 8242322 TI - Laparoscopic abdominoperineal resection of the rectum. PMID- 8242323 TI - Conventional versus laparoscopic surgery for acute appendicitis. PMID- 8242324 TI - Contemporary management of appendiceal mass. PMID- 8242325 TI - Single-loop biliary and gastric bypass for irresectable pancreatic carcinoma. PMID- 8242326 TI - Prospective randomized study of one- or two-layer anastomosis following oesophageal resection and cervical oesophagogastrostomy. PMID- 8242327 TI - Laparoscopic mobilization and exteriorization for minimally invasive small bowel resection. PMID- 8242328 TI - Randomized trial comparing three forms of pelvic floor repair for neuropathic faecal incontinence. PMID- 8242329 TI - Regional anaesthesia and surgical morbidity. PMID- 8242330 TI - Effect of suture material on tumor cell adherence at sites of colonic injury. PMID- 8242331 TI - Vibration white finger. PMID- 8242332 TI - Return to work after inguinal hernia repair. PMID- 8242333 TI - Localization of neurotrophin-3-like immunoreactivity in peripheral tissues of the rat. AB - Neurotrophin-3 (NT-3) mRNA is widely distributed in both the peripheral and central nervous systems but neither the distribution of the native factor nor its physiological function is known. In the present study we produced and characterized an antibody to a synthetic peptide and showed that it specifically recognised endogenous rat and recombinant human NT-3 (rNT-3), but not mouse nerve growth factor and recombinant brain derived-neurotrophic factor. NT-3-like immunoreactivity (NT-3-ir) was detected within the distal tubular cells of the kidney, the zona glomerulosa and reticularis of the adrenal cortex, ganglion cells of the adrenal medulla, red pulp of the spleen, epithelial cells of the intestinal villi and parenchymal cells of the liver. Within peripheral ganglia, NT-3-ir was observed in a subpopulation of large sensory neurons of dorsal root, trigeminal and cochleovestibular ganglia but not in principle neurons of the sympathetic ganglia. These results provide the first evidence for the localization of NT-3-ir and indicate its presence in various peripheral organs and large sensory neurons. We conclude that NT-3 may function outside the nervous system in addition to a neurotrophic role within large sensory neurons. PMID- 8242334 TI - Effects of castration, estrogen replacement and estrus cycle on monoamine metabolism in the nucleus accumbens, measured by microdialysis. AB - Changes in monoamine concentration in the nucleus accumbens were measured by microdialysis during the estrus cycle of intact females rats as well as untreated ovariectomized rats and ovariectomized rats treated with estrogen and progesterone. Norepinephrine (NE), 3-methoxy-4-hydroxyphenylglycol (MHPG) dopamine (DA), 3,4-dihydroxyphenylacetic acid, (DOPAC), serotonin (5-HT) and 5 hydroxy-3-indoleacetic acid (5-HIAA) were measured by high performance liquid chromatography. The ratio of MHPG to NE and DOPAC to DA were significantly increased during proestrus when compared to other phases of the estrus cycle. The ratio of 5-HIAA:5-HT, however, was significantly reduced during proestrus. Treatment of ovariectomized rats with estradiol at either 1 microgram/day or 10 micrograms/day significantly increased the ratio of MHPG:NE and DOPAC:DA. In contrast, the ratio of 5-HIAA:5-HT decreased significantly. Progesterone had no significant effect on the concentrations of 5-HT or 5-HIAA, but attenuated the effect of estrogen on the ratio of MHPG:NE and similarly reduced the ratio of DOPAC:DA. These studies are consistent with the hypothesis that estrogens are associated with enhanced metabolism of NE and decreased metabolism of 5-HT and dopamine in the nucleus accumbens of freely moving rats. PMID- 8242335 TI - L-BOAA induces selective inhibition of brain mitochondrial enzyme, NADH dehydrogenase. AB - Lathyrism, a human neurological disorder has been linked to the excessive consumption of a plant toxin, beta-oxalylamino-L-alanine (L-BOAA) present in Lathyrus sativus. The present study was carried out to elucidate the biochemical mechanisms underlying L-BOAA-induced toxic insult. Incubation of sagittal slices of mouse brain with L-BOAA resulted in dose and time-dependent inhibition of mitochondrial NADH-dehydrogenase (NADH-DH). Significant inhibition of NADH-DH was seen following incubation of brain slices with very low concentration of L-BOAA (0.1 pM). L-BOAA also induced lactate dehydrogenase (LDH) leakage from the slice into the medium in dose-dependent manner. The inhibition of NADH-DH preceded LDH leakage from the slices into the medium. L-BOAA had no effect on other mitochondrial enzymes, namely, isocitrate dehydrogenase or cytochrome c oxidase. Incubation of isolated mouse brain mitochondria with L-BOAA also resulted in inhibition of NADH-DH. L-BOAA-induced inhibition of NADH-DH was prevented by non N-methyl-D-aspartate (non-NMDA) glutamate receptor antagonists in general and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor antagonist (NBQX) in particular. Other glutamate agonists examined namely, N-methyl-D aspartate, beta-N-methylamino-L-alanine (L-BMAA), L-glutamic acid, N acetylaspartylglutamate (NAAG), quisqualic acid, kainic acid or AMPA did not have any effect on NADH-DH activity in slices although they induced LDH leakage from the slice into the medium. Incubation of brain slices with L-BOAA did not induce lipid peroxidation or changes in glutathione levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242336 TI - Expression of cathepsin G-like and alpha 1-antichymotrypsin-like proteins in reactive astrocytes. AB - The central nervous system (CNS) of many different species responds to diverse neurologic injuries with an activation of astrocytes. Yet, the exact function of this reactive astrocytosis is unknown. In this report, mouse astrocytes were activated in vivo by focal penetrating brain injury. Reactive astrocytes were stained with antibodies raised against the serine protease cathepsin G (cat.G), the serine protease inhibitor alpha 1-antichymotrypsin (ACT), or the astrocytic marker glial fibrillary acidic protein (GFAP). Reactive astrocytes expressing both cat.G-like and ACT-like antigens were found around cerebral wound margins between 18 h and 13 days after neural lesions. The injury-induced immunostaining was unaltered by 900 rads of total body irradiation, suggesting that the astroglial reaction was relatively independent of bone marrow-derived cells. The in vivo immunostaining was complemented with biochemical assays on cultured primary astrocytes. A synthetic peptide was used as a substrate in combination with specific inhibitors to identify a proteolytic activity within astroglial lysates and culture supernatants that closely resembles cat.G. This activity increased substantially upon stimulation of astrocytes with dibutyryl cyclic AMP and was neutralized by antibodies raised against cat.G. In a separate report, it was shown that astrocytes also contain an ACT-like inhibitory activity. The production of ACT- and cat.G-like antigens and activities by activated astrocytes should allow these cells to participate in a number of important biologic processes. Many of these processes may benefit the CNS by assisting in early wound repair. However, astroglial proteases and their inhibitors could also contribute to the pathogenesis of certain neurologic diseases. PMID- 8242337 TI - Cytokine stimulation increases intracellular calcium and alters the response to quisqualate in cultured cortical astrocytes. AB - Cytokine levels are elevated in the central nervous system (CNS) in a variety of disorders and may contribute to abnormalities in CNS function associated with the disorders. To begin to understand the mechanisms through which elevated cytokines affect CNS cells, we have examined the effects of cytokines on astrocyte physiology within minutes of application as well as 24 h later. Both standard cultured cortical astrocytes and those induced to further differentiate by pre treatment with forskolin were examined. Such treated astrocytes may more closely resemble those in brains exhibiting elevated cytokine levels. The cytokine focused upon was interleukin-1-beta (II-1 beta). Gamma-interferon (gamma-IFN) and tumor necrosis factor-alpha (TNF-alpha) were also examined in some studies. Changes in calcium levels produced by acute application of these cytokines were measured. The most pronounced effect was an immediate calcium elevation in response to II-1 beta in the forskolin pre-treated astrocytes. Longer term treatment with IL-1 beta in forskolin pre-treated astrocytes enhanced the calcium response to quisqualate stimulation, a glutamate neurotransmitter receptor agonist. These results suggest that situations that cause chronic changes in cytokine levels and involve astrocytic differentiation, such as chronic CNS infection or Alzheimer's disease, could change astrocytic responses to normal stimuli. Such changes may result in altered astrocytic support of neurons and therefore cause changes in CNS function. PMID- 8242338 TI - Susceptibility of human foetal brain tissue to cool- and freeze-storage. AB - Human second trimester foetal brain tissue was stored for a period of 1-6 weeks under various conditions in an attempt to evaluate factors influencing its susceptibility (cell loss) and survivability. Post-storage viability of mesencephalon, striatum, cerebellum and occipital cortex was assessed by a protocol combining vital staining with cell density counts so that tissue viability and cell loss could be evaluated simultaneously; tissue survivability was evaluated by cell culture. A significant amount of cell loss occurred after 24 h storage at room temperature, after one week at 4 degrees C and by two weeks at -20 degrees C in all structures; storage at -196 degrees C resulted in 17-21% cell loss at the end of a 6 week period. At -20 degrees C the cryoprotective effect of 20% FCS was equivalent to that of 15% FCS + 7% DMSO combined, suggesting potential use of serum in replacement of chemical additives. The procedure for removal of DMSO was critical to cell viability and survivability: single step dilution led to 27-39% greater cell loss than slow, multi-step dilutions. In comparison to fresh, non-stored tissue, immunocytochemical characterization of in vitro propagated stored tissue revealed no changes in the populations of major constituent cell types including neurones, dopaminergic neurones, glial and fibroblast cells. These results provide information on possible conditions under which transplant tissue can be satisfactorily stored depending on the prevailing requirements. PMID- 8242339 TI - Age-related reduction of human growth hormone-binding sites in the human brain. AB - Previous studies have shown that alterations in various neuroendocrine functions occur with increasing age. We here report a study of growth hormone (GH)-binding sites in different areas of post-mortem human brains collected from individual males and females of different age. The results indicate that there exists a significant negative correlation between the density of GH-binding sites and increasing age. This phenomenon was observed in both sexes in brain areas such as choroid plexus, hippocampus, hypothalamus, pituitary and putamen but not in e.g. thalamus. In all tissues (except for choroid plexus), the GH binding was significantly higher in those originating from females than those from males. This discrepancy was found likely to be associated with the affinity of GH to lactogenic rather than to somatogenic sites as no pronounced sex difference in binding was observed in the presence of excessive amounts of human prolactin. Data also indicate that the putative GH receptors in the various brain regions differ with regard to binding constants and to the estimated molecular size of the hormone-binding units. The loss of GH receptors in brain of elderly people may have consequences in several physiological courses. The decrease in GH binding at hypothalamic and pituitary levels may be of importance for the mechanisms behind the release or secretion of the hormone. PMID- 8242340 TI - Release of amino acids into regional superfusates of the spinal cord by mechano stimulation of the reproductive tract. AB - Based on pharmacological evidence that inhibitory amino acids mediate vaginocervical mechano-stimulation produced analgesia (VSPA), we hypothesized that inhibitory amino acids would be released endogenously in the spinal cord in response to vaginocervical mechano-stimulation (VS). This hypothesis was tested by HPLC analysis of the amino acid content of 5-min superfusates of the spinal cord before, during and after VS (400 g force applied against the cervix) in urethane-anesthetized rats. Utilizing an in vivo push-pull superfusion method, artificial cerebrospinal fluid was continuously superfused over the spinal cord through the intrathecal space surrounding the sacral-lower thoracic region. In addition, concentrations of amino acids in the superfusate were measured in response to KCl stimulation (increasing the superfusion medium from 3.4 to 40.0 mM KCl to produce non-specific depolarization), and noxious hind paw mechano stimulation (pinching the hind paw to produce a sustained flexor response in ipsilateral hind leg). There was a significant increase in the concentration of Gly, Tau, Asp, Glu and Lys in the superfusate in response to VS (n = 8) and to KCl (n = 8), but not to hind paw stimulation (n = 5). Also, GABA concentrations increased in response to KCl, and the concentration of Ala, Ser, Gln, Thr, Arg and Phe increased in response to VS, however, GABA levels were sometimes below the limits of detection. In contrast, there was no significant change in any amino acid concentration in response to hind paw pinch stimulation, and VS did not significantly affect the concentrations of Tyr, His, Ile, Leu, Met, Trp or Val. The present findings support our hypothesis that VS releases inhibitory amino acids in the spinal cord. Moreover, other amino acids, including 'excitatory' amino acids, are released into the superfusate. The profile of amino acid release in response to VS differs from that in response to paw pinch or KCl administration. PMID- 8242341 TI - Nimodipine prevents the in vivo decrease in hippocampal extracellular acetylcholine produced by hypobaric hypoxia. AB - Hypoxia decreases acetylcholine (ACh) synthesis and release in vitro, and ACh synthesis in vivo; however, its effect on extracellular concentration of ACh in vivo is not known. The calcium channel blocker nimodipine is a cerebrovascular dilator which also increases extracellular ACh in vivo. Therefore, it may provide protection from the effects of hypobaric hypoxia on the cholinergic system either via its effects on vascular function or by direct action on the nervous system. This study examined the effect of hypobaric hypoxia on extracellular ACh and choline levels, as measured by microdialysis, as well as the effects of nimodipine under hypoxia. Microdialysis guide cannulae were implanted into the hippocampal region of male Fischer rats so that probes would sample from the CA1 and DG regions. Animals were then exposed for eight hours to a simulated altitude of 5,500 m (18,000 ft) or tested at sea level for an equivalent duration. HPLC with electrochemical detection was used for analysis of the dialysates. At 5,500 m extracellular ACh levels in the placebo-treated group were significantly lower than the sea level group values. This decrement was reversed by nimodipine administered i.p. immediately preceding altitude ascent (10 mg/kg) and 250 min post-altitude ascent (10 mg/kg). These data suggest that nimodipine may provide protection from the detrimental effects of hypoxia on hippocampal cholinergic function. PMID- 8242342 TI - Effects of discrimination learning on the rat amygdala dopamine release: a microdialysis study. AB - Extracellular levels of dopamine (DA) and its metabolites were monitored with the in vivo microdialysis method in the amygdala of rats while and after they performed a discrimination learning task or a non-discrimination task. A group of rats was trained to discriminate between lamp-on and lamp-off states under an operant-type learning procedure. After a stable discriminative behavior was established, a dialysis probe was inserted into the basolateral amygdaloid nucleus of each rat. The concentrations of DA and its metabolite 3,4 dihydroxyphenylacetic acid significantly increased during the learning sessions as compared to their basal levels. In contrast, another group of rats trained on a similar, but non-discriminative task showed no such increases. These results suggest that the dopaminergic neural systems in the basolateral amygdaloid nucleus are activated during ongoing behavior maintained under the discrimination learning situation. PMID- 8242343 TI - Widespread expression of gephyrin, a putative glycine receptor-tubulin linker protein, in rat brain. AB - The peripheral membrane protein gephyrin co-purifies with the inhibitory postsynaptic glycine receptor (GlyR) of mammalian spinal cord. By immunoelectron microscopy, gephyrin has been localized at the cytoplasmic face of glycinergic postsynaptic membrane specializations. Here, we used specific monoclonal antibodies to demonstrate the presence of gephyrin in all regions of rat brain known to contain synapses and compared its histochemical distribution with that of GlyR antigens. In most brain structures, gephyrin is expressed independently of the GlyR alpha 1 subunit, but its distribution is very similar to the pattern obtained with mAb 4a, a monoclonal antibody recognizing the known GlyR alpha and beta subunits. Our data suggest a much wider distribution of gephyrin and GlyR proteins in the mammalian CNS than anticipated previously. PMID- 8242344 TI - Effects of systemic nicotine on serotonin release in rat brain. AB - We used in vivo microdialysis to examine the acute effects of systemically administered nicotine (0.8-8.0 mg/kg, s.c.) on extracellular levels of serotonin (5-HT) in the frontal cortex of awake rats and animals anesthetized with chloralose/urethane. In anesthetized animals, 5-HT efflux was elevated during the initial 15 min after nicotine administration (2-8 mg/kg), but then returned to baseline values. All of the effective nicotine doses also lowered and then raised blood pressure in these animals. However, other drugs which raised (methoxamine, 0.07 mg/kg, i.v.) or lowered (mecamylamine, 5 mg/kg, i.p.) blood pressure without directly activating nicotinic receptors failed to alter 5-HT release. Moreover, pretreatment with a centrally active dose of mecamylamine, a known nicotinic antagonist, blocked the effects of nicotine (4 mg/kg) on 5-HT release. For studies on awake rats the perfusion fluid also contained fluoxetine, since basal 5-HT levels were barely detectable without this uptake blocker. In such animals, 1.6 mg/kg of nicotine significantly increased 5-HT release, an effect apparent in the initial 20 min after treatment and persisting for at least 2 h. These observations demonstrate that systemically administered nicotine increases frontocortical 5-HT release, that this effect is independent of the cardiovascular responses to the drug, and that it probably results from the activation of previously described nicotinic receptors on raphe neurons. The present findings are consistent with the hypothesis that the appetitive and mood disturbances associated with nicotine withdrawal may be mediated by diminished serotoninergic transmission. PMID- 8242345 TI - Disturbances in the performance of thermal discrimination tasks following cortical ablations in rats. AB - Previous studies of the effects of ablating the rat's somatosensory cortex on temperature discrimination have yielded negative results. Presently, it was assumed that (1) the rat's face might possess thermal acuity comparable to that found in highly sensitive skin regions of primates, (2) the rat's facial discriminative capacity for thermal differences might be more acute in a cool range well below normal room temperature (24 degrees C), and (3) by using more sensitive procedures and focusing on the effects of damage to the face areas in the rat's somatosensory cortex, disturbances in the capacity to make discriminations between cool stimuli might be revealed that previously went unnoticed. Results of experiments testing these assumptions indicated that rats can use their snouts to make discriminations of 1 degree C or less, that their acuity is better in the cool than in the warm range, and that somatosensory ablations produce moderate to severe disturbances in the capacity to discriminate between cool stimuli but only slight transitory disturbances in this capacity for warm stimuli. Additionally, the results suggest that the sensorimotor cortex may be involved in the rat's thermal discriminative capacity. PMID- 8242346 TI - Nerve growth factor synthesis by mouse submandibular gland cells in culture. AB - Mouse submandibular gland (SMG) cells in culture rarely retain functional properties of SMG cells in vivo. We demonstrate that both primary SMG cells and the mouse SMG cell line SCA-9 secrete biologically active nerve growth factor (NGF). However, primary cells secrete 40-fold more NGF than SCA-9 cells, demonstrating that SCA-9 cells cannot substitute for primary SMG cells for the study of SMG NGF in vitro. PMID- 8242347 TI - Neuropeptide Y in the arcuate nucleus is modulated by alterations in glucose utilization. AB - This study examined the response of hypothalamic neuropeptide Y (NPY) to specific metabolic challenges. After intraperitoneal administration of 2-deoxy-D-glucose, which blocks glucose utilization, NPY levels measured via radioimmunoassay were significantly potentiated in the arcuate (ARC) and suprachiasmatic nuclei of the rat hypothalamus. The antimetabolite mercaptoacetate, in contrast, which blocks fatty acid oxidation, produced no significant change and actually tended to reduce NPY levels in the ARC. It is concluded that glucose utilization, in particular, may constitute an important signal, either direct or indirect, in the modulation of NPY production in the hypothalamus. PMID- 8242348 TI - Increased activities of MAOA and MAOB in autopsied brain tissue from cirrhotic patients with hepatic encephalopathy. AB - Using radioenzymatic assays, activities of MAOA and MAOB were measured in autopsied brain tissue from cirrhotic patients who died in hepatic coma and in material from an equal number of age-matched subjects who were free from hepatic, neurological or psychiatric disorders. Activities of both MAOA and MAOB were significantly increased in frontal cortex and caudate nucleus, two brain regions shown previously to be the site of functional and morphological alterations of astrocytes and increased concentrations of the acid metabolites of dopamine and serotonin. These findings suggest that increased monoamine metabolism and subsequent modifications of monoaminergic synaptic function could contribute to the pathogenesis of hepatic encephalopathy. PMID- 8242349 TI - Distribution of GABA-like immunoreactivity in the octopus brain. AB - This paper presents the first evidence that some neurons in the octopus CNS contain delta-amino butyric acid (GABA). Using conventional immunohistochemical methods with appropriate controls, we obtained positive staining with an antibody to GABA in fibres in the neuropil of many lobes of the brain of the northern octopus Eledone cirrhosa. In several lobes cell bodies were also stained. Staining was not uniformly distributed in the brain nor within a particular lobe: some regions stained strongly, others not at all. These findings suggest that GABA should be added to the already long list of putative neurotransmitters in the cephalopod CNS. PMID- 8242350 TI - Neurotensin injected into the nucleus accumbens blocks the psychostimulant effects of cocaine but does not attenuate cocaine self-administration in the rat. AB - The neuropeptide neurotensin (NT) has been shown to modulate mesolimbic dopaminergic activity. Neurotensin injected into the VTA produces motor stimulation and release of dopamine in the nucleus accumbens. In contrast, when neurotensin is administered into the nucleus accumbens, it produces neuroleptic like effects such as attenuation of the locomotor activity elicited by psychostimulants. In the present study, the hypothesis that neurotensin injected into the nucleus accumbens might modulate the psychostimulant and reinforcing actions of cocaine was tested. In experiment one, rats were trained to self administer cocaine intravenously on an FR5 schedule of reinforcement. Following the establishment of baseline responding, rats were implanted with bilateral cannulae in the nucleus accumbens. One week later, rats were injected into the nucleus accumbens with various doses of neurotensin (4.2, 8.4 and 16.7 micrograms, total doses bilaterally) immediately prior to the self-administration session. No significant effects were found with any of the doses of neurotensin tested on the self-administration of cocaine. However, in experiment 2, neurotensin at doses of 4.2 and 16.7 micrograms injected into the nucleus accumbens significantly reduced the locomotor activation induced by an acute injection of cocaine (15 mg/kg i.p.) and a dose of 16.7 micrograms attenuated the locomotor activation induced by amphetamine (0.75 mg/kg i.p.). Thus, neurotensin in the nucleus accumbens appears to specifically modulate the acute locomotor activating properties of cocaine but not cocaine self-administration. Different mechanisms by which NT interacts with dopamine in the nucleus accumbens may provide a means of selectively altering psychostimulant motor actions without affecting psychostimulant reinforcement. PMID- 8242351 TI - Brain cytochrome oxidase activity after kindled seizures: a quantitative histochemical mapping study. AB - Quantitative histochemistry was used to analyze changes in cytochrome oxidase (CO) activity in 93 brain regions after entorhinal cortex kindling. Rats were kindled to at least six stage-5 seizures and sacrificed either 24 h or 28 days after the last convulsion. Regional brain CO activity was quantitated in histological sections using calibrated densitometric standards. No statistically significant differences in regional CO activity between kindled and control brains were seen either 24 h or 28 days after the last convulsion. These results suggest that the brain changes underlying the kindling state are not reflected in localized alterations in mitochondrial respiratory capacity. They also indicate that long-lasting changes in regional brain CO activity recently found after electroconvulsive shock are not common to all types of seizures. PMID- 8242352 TI - Characterization of metabotropic glutamate receptors negatively linked to adenylyl cyclase in brain slices. AB - We have characterized the pharmacological profile of activation of metabotropic glutamate receptors negatively linked to adenylyl cyclase (mGluR decreases cAMP) in brain slices. Among the putative mGluR agonists, (2S,1'R,2'R,3'R)-2-(2,3 dicarboxycyclopropyl)glycine (DCG-IV) and (1S,3R)-1-aminocyclopentane-1,3 dicarboxylic acid (ACPD), were the most potent inhibitors of forskolin-stimulated cAMP formation in hippocampal slices, followed by ibotenate, L-2-amino-3 phosphonopropionate (AP3), quisqualate, L-glutamate and beta-N-methylamino-L alanine (BMAA). Inhibition of forskolin-stimulated cAMP formation by DL-2-amino-4 phosphonobutanoate (AP4) was biphasic, suggesting that the drug interacts with more than one mGluR decreases cAMP subtype. Both L-AP4 and L-serine-O-phosphate (a restricted analogue of AP4) were much more effective in inhibiting forskolin stimulated cAMP formation than their D-isomers, indicating that interaction of these drugs with the mGluR decreases cAMP is stereoselective. Despite the fact that DCG-IV and ibotenate behave as NMDA receptor agonists, their effect was insensitive to MK-801. The regional pattern of expression of mGluR decreases cAMPS, as estimated by using 1S,3R-ACPD as an agonist, did not correlate with the steady-state levels of mGluR2 mRNA. Thus, 1S,3R-ACPD inhibited forskolin stimulated cAMP in slices from hippocampus, cerebral cortex, corpus striatum, olfactory tubercle or hypothalamus, but not in slices from olfactory bulb or cerebellum; in contrast, mGluR2 mRNA levels were high in the olfactory bulb and very low in the corpus striatum. 1S,3R-ACPD also inhibited forskolin-stimulated cAMP formation in cortical membranes, excluding the involvement of trans-synaptic mechanisms in the activity of mGluR decreases cAMPS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242353 TI - Pharmacological consequences of nicotinergic plus serotonergic manipulations. AB - The present study investigates the effects of concurrent manipulations of nicotinic cholinergic receptors (nicotinic cholinergic agonist: nicotine 0.03, 0.1, 0.3 mg/kg, nicotinic cholinergic antagonist: mecamylamine 7.5 mg/kg) and serotonin neurons (p-chlorophenylalanine (PCPA), 400/kg mg on each of 3 days) on spatial navigation (water maze, WM) and passive avoidance (PA) performance. Nicotine did not affect PA performance but at the highest dose slightly impaired WM performance. PCPA did not affect WM navigation or PA performance in saline or nicotine-treated rats. Nicotine restored WM and PA performance defect in mecamylamine pretreated rats. PCPA aggravated the WM defect and decreased the WM performance-improving effect of nicotine in mecamylamine pretreated rats. PCPA did not aggravate the PA performance defect of mecamylamine but completely blocked the PA performance-improving effect of nicotine in mecamylamine pretreated rats. These results suggest that serotonergic and nicotinergic cholinergic systems jointly modulate performance in WM and PA tests. PMID- 8242354 TI - Voltage gated calcium channel currents of rat dorsal root ganglion (DRG) cells are blocked by Al3+. AB - The effects of the trivalent cation aluminum (Al3+) on voltage activated calcium channel currents were examined. Al3+ blocks sustained and transient components of voltage activated calcium channel currents of cultured rat dorsal root ganglion (DRG) cells. Currents were elicited by voltage jumps from -80 to 0 mV. The channel block was use dependent. Steady state blockade occurred after 1 to 5 min, when opening the channel every 10 s. There was little or no recovery after washing. Threshold concentration was about 20 microM Al3+ and total blockade (> 80%) was obtained at 200 microM Al3+; the IC50 was 83 microM and the Hill number was around 3. The degree of blockade was pH dependent, and increased with H+ concentration. The current-voltage relation frequently shifted to depolarised voltages after applying Al3+. The degree of the shift was a function of Al3+ concentration, but the magnitude differed from cell to cell. In the effective concentration range (< 200 microM Al3+) the effect was quite specific to voltage activated calcium channel currents. Voltage activated potassium and sodium channels were reduced less than 15% by 200 microM Al3+. We conclude that Al3+ is a potent and irreversible blocker of voltage activated calcium channel currents in mammalian neurons. PMID- 8242355 TI - The cholinergic receptor-linked phosphoinositide metabolism in mouse cerebrum and cerebellum in vivo. AB - The cholinergic receptor-linked poly-phosphoinositide hydrolysis was studied in mouse cerebrum and cerebellum after prelabeling the brain with [3H]inositol. I.p. injection of Li (8 meq/kg) to C57Bl/6J mice for 4 h resulted in 14- and five-fold increases in [3H]inositol-labeled inositol monophosphate (IP1) in cerebrum and cerebellum, respectively. The labeled inositol bisphosphate (IP2) was also increased 83 and 19% in cerebrum and cerebellum, respectively. Prior injection of atropine (100 mg/kg) resulted in inhibition of Li-induced increases in labeled IP1 by 74 and 56% in cerebrum and cerebellum, respectively. Administration of pilocarpine (20 mg/kg) to the Li-treated mice for 30 min resulted in further increases in labeled IP1 and IP2 and a concomitant decrease in labeled inositol in cerebrum but not in cerebellum. Mass measurements of IP1 and IP2 isomers by HPLC revealed that inositol 1-monophosphate (Ins(1)P), inositol 4-monophosphate (Ins(4)P) and inositol 1,4-bisphosphate (Ins(1,4)P2) were all increased by pilocarpine administration in the Li-treated mouse cerebrum. The effects of pilocarpine administration in mouse cerebrum (increases in IP1 and IP2) could be completely inhibited by preinjection of atropine. Atropine injection also decreased the levels of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3]. Surprisingly, a decrease in Ins(1,4,5)P3 level was also found in non-Li-treated mice after pilocarpine administration (30 mg/kg, 10-40 min). Except for the increase (20%) in [32P]-labeled PIP in the cerebrum, Li or Li together with pilocarpine administration did not alter the levels of [3H]inositol or [32P]phosphate-labeled phosphoinositides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242356 TI - Comparison between the membrane and synaptic properties of human and rodent dentate granule cells. AB - We have compared the cellular and synaptic properties of rodent dentate granule cells with those of humans. The human tissue was obtained from neurosurgical procedures which necessitated removal of the hippocampus for treatment of extra hippocampal tumors which presented clinically with seizures. The hippocampi studied here were neuroanatomically similar to autopsy controls. The present studies have demonstrated that there are few differences between rodent and human granule cells as regards either their membrane properties or their synaptic physiology and pharmacology. The differences we noted were (1) less spike frequency adaptation in the human relative to rodent cells; and (2) perforant path stimulation reliably elicited both feedforward and feedback inhibition in the rodent cells, while in the human tissue feedback inhibition appeared to predominate. It is unclear if these changes are due to the seizure experience or if they represent true species differences. PMID- 8242357 TI - Abnormal distribution of acetylcholinesterase activity in the hippocampal formation of the dreher mutant mouse. AB - The distribution of acetylcholinesterase(AChE) in the hippocampal formation of the dreher mutant mouse was studied by comparing homozygous mutant (drsst-J/drsst J) with littermate control (+/? or +/+). In the control mice, AChE activity was most intense in the inner one-third of the stratum oriens and lacnosum of the hippocampus, and in the inner one-fifth of the molecular layer of the dentate gyrus. In contrast, in homozygous dreher mice, AChE activity in area CA3c of the hippocampus was not restricted to the stratum oriens, and extended upward into the infrapyramidal and suprapyramidal mossy fiber layers, the lower part of the stratum radiatum, the pyramidal cell layer, and downward toward the alveus. In addition, the distribution of AChE activity was modified by accompanying with ectopic pyramidal cells or with disruption of the pyramidal cell layer. AChE activity in the dentate gyrus of the dreher mouse was not confined to the inner one-fifth of the molecular layer. These findings indicated that the cholinergic input to the hippocampal formation is not normal in the dreher mutant mouse. Since the areas of AChE activity correspond to the presence of ectopic pyramidal cells in the dreher mouse, incoming cholinergic fibers may form synapses with these ectopic cells and with the dendrites of normal pyramidal cells that extend into the expanded area of AChE activity. PMID- 8242358 TI - The receptors activated by exogenous and endogenous opioids in the superior cervical ganglion of the cat. AB - Nicotinic transmission in the superior cervical ganglion of the cat was studied in vivo and in vitro by recording the postganglionic compound action potential evoked, under partial block with hexamethonium, by supramaximal 0.2 Hz stimulation of the cervical sympathetic trunk. The compound action potential was depressed following a stimulus train (5 Hz, 40 s) applied to the same set of axons. The inhibition was antagonized by naloxone, suggesting mediation by endogenous opioids. Agonists selective for mu-, delta- and kappa-opioid receptors, injected into the arterial supply of the in situ ganglion or added to the perfusion fluid in the in vitro ganglion, also produced a naloxone-sensitive inhibition of the compound action potential. The synaptic inhibition was antagonized by the delta-selective antagonist ICI 174,864 and by the mu-selective antagonist CTAP, but not by the kappa-selective antagonist Nor-BNI. These results suggest that all three main types of opioid receptors are present in the ganglion but only mu- and delta-receptors are involved in the inhibition of ganglionic transmission mediated by the endogenous opioids. PMID- 8242359 TI - Response properties of hypogastric afferent fibers supplying the uterus in the cat. AB - The present study was undertaken to examine the sensory function of uterine afferent fibers in cats at unknown stages of the estrous cycle. Single unit activity was recorded from strands of the hypogastric nerve of the anesthetized cat. Once a unit was found, the conduction velocity was determined and the mechanical receptive field localized on the uterus. The response properties of the unit to mechanical stimuli applied to the receptive field and to chemical stimuli applied by intra-arterial injection of algesic chemicals (bradykinin, KCl and capsaicin) into the uterine artery were studied. Single unit activity from a total of 52 units was examined in this study. Based on the conduction velocities, about 2/3 of these fibers were found to be unmyelinated C fibers and the remaining 1/3 were thinly myelinated A delta fibers. The receptive fields of most of these fibers were located at different parts of the uterine horn and body while a few were at the uterine cervix. Mechanical thresholds, as determined by von Frey filaments applied to the external surface of the uterus, varied more than 150-fold among mechanically sensitive units, ranging from extremely low to high thresholds. In addition, most of these afferents were activated by intra arterially injected algesic chemicals, often by more than one chemical. The data in the present study suggest that a large portion of the cat uterus is innervated by the hypogastric nerve and that these afferents originate from sensory receptors that have potentially a wide range of functions. Their potential functions as low threshold mechanoreceptors and nociceptors are discussed. PMID- 8242360 TI - Up-regulation of androgen receptor immunoreactivity in the rat brain by androgenic-anabolic steroids. AB - To characterize central nervous system changes that occur with anabolic steroid abuse in humans, immunocytochemical localization of androgen receptors in the brains of 10 intact and 10 castrated male rats was conducted after the administration of high levels of androgenic-anabolic steroids (AAS; 14 daily injections of sesame oil or a cocktail of 2 mg/kg testosterone cypionate, 2 mg/kg nandrolone decanoate, and 1 mg/kg boldenone undecylenate). In normal intact oil treated males, nuclear androgen receptor immunoreactivity was present in many 'classical' and 'non-classical' androgen target sites in the brain. Administration of AAS increased the intensity of immunoreactivity in most classical androgen target sites and increased both the intensity of immunoreactivity and number of immunoreactive cells in most non-classical androgen target sites. These results may suggest that androgen receptors in the brain are up-regulated by AAS. The simultaneous androgen receptor up-regulation in these regions by AAS may account for the complex anabolic steroid abuse syndrome. Consistently, androgen receptor immunoreactivity in most brain regions was reduced or absent after castration, suggesting that endogenous androgen levels are necessary for normal androgen receptor immunoreactivity. These results identify the distribution of one central nervous system mechanism modified by AAS. PMID- 8242361 TI - Use of 51Cr cell labelling to distinguish between release of radiolabelled amino acids from primary astrocyte cultures being due to efflux or cell damage. AB - Continuous perfusion methods are widely used to monitor release of substances, particularly transmitters, from brain cell cultures growing as monolayers. However, if stimuli used to produce release also cause loss or lysis of cells, the appearance of label in the perfusate due to such effects will be indistinguishable from release. Using a perfusion method we have studied release of preloaded, radiolabelled amino acids from primary astrocyte cultures due to a variety of stimuli; hypotonic or high K+ media, activation of beta-receptors or swelling-induced release due to isosmotic ethanol. In this study primary astrocyte cultures were simultaneously labelled with Na2(51)CrO4 and allowed to take up radiolabelled D-aspartate or taurine. It was found that while all of the above methods caused release of radiolabelled amino acids none caused release of 51Cr into the perfusion fluid. In contrast, perfusion with 0.05% (v/v) Triton X 100 did lead to release of 51Cr. Thus a variety of means of inducing swelling or shape changes in astrocytes causes true release of radiolabelled amino acids and simultaneously monitoring 51Cr release seems a convenient means of distinguishing such release from cell loss or lysis. PMID- 8242362 TI - Synergism between androgens and estrogens in the induction of aromatase and its messenger RNA in the brain. AB - It is established that testosterone (T) increases aromatase activity (AA) in the quail brain and that this induction of AA represents a limiting factor in the activation of male copulatory behavior. This action of T presumably results from an induction of aromatase synthesis since the number of aromatase-immunoreactive (ARO-ir) cells increases and, in parallel, there is an increase in aromatase mRNA as measured by reverse transcriptase-polymerase chain reaction (RT-PCR) technology. The specific role of androgenic and estrogenic metabolites of T in this induction is not yet clear but product-formation assays suggest that both types of compounds synergize to increase AA. The exact role of androgens and estrogens in the induction of aromatase was examined by studying both the aromatase protein by immunocytochemistry and the aromatase mRNA by RT-PCR in castrated quail that had been treated with T or its androgenic metabolite, 5 alpha-dihydrotestosterone (DHT) or its estrogenic metabolite, estradiol-17 beta (E2) or both DHT and E2 simultaneously. A specific quantitative PCR technique using a modified aromatase as internal standard was developed for this purpose. T increased the number of ARO-ir cells in all brain areas and increased the concentration of ARO mRNA in the preoptic area-anterior hypothalamus (POA-aHYP) and in the posterior hypothalamus (pHYP). E2-treated birds had more ARO-ir cells than castrates in the posterior part of the medial preoptic nucleus (POM), in the bed nucleus stria terminalis (BNST) and tuber. Their aromatase mRNA concentration was significantly increased in the POA-aHYP but this effect did not reach significance in the pHYP. DHT by itself had no effect on either the number of ARO ir cells (all brain regions considered) or the concentration of aromatase mRNA. DHT, however, synergized with E2, both in inducing ARO-ir neurons and in increasing aromatase mRNA concentration. This synergism was shown to be statistically significant in several brain areas. These data demonstrate that both androgens and estrogens regulate aromatase at the pretranslational level. Because the percentage increase in the number of ARO-iR cells was in general very similar to the increase in aromatase mRNA concentration, these data also suggest that these steroids regulate aromatase mostly by changing its mRNA synthesis or catabolism. PMID- 8242363 TI - Cytokine-induced change in hypothalamic norepinephrine turnover: involvement of corticotropin-releasing hormone and prostaglandins. AB - Changes in norepinephrine (NE) turnover in restricted brain regions were examined in rats after administration of the major mediators of the acute phase response, interleukin-1 beta (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF). An increase in NE turnover was observed after intraperitoneal injection of IL-1 (1 microgram/rat) in the whole hypothalamus and several specific hypothalamic nuclei, but not in the medulla oblongata and cerebral cortex. The stimulatory effect of IL-1 was mimicked by an intracerebroventricular injection of much lower doses of IL-1 (10-100 ng/rat). This IL-1-induced increase in hypothalamic NE turnover was blocked by the pretreatment with either indomethacin (cyclooxygenase inhibitor) or anti-corticotropin releasing hormone (CRH) antibody but not by naloxone. Intracerebroventricular injection of CRH increased NE turnover not only in the hypothalamus but also in the medulla oblongata and cerebral cortex. However, prostaglandin (PG) E2 and PGF2 alpha did not show such effect. It was therefore suggested that IL-1 activates noradrenergic neurons projecting to the hypothalamus by its direct action to the brain, and that CRH and eicosanoid-cyclooxygenase product(s) within the brain are involved in this process. In contrast, neither IL-6 nor TNF influenced brain NE turnover regardless of whether they were given intraperitoneally or intracerebroventricularly. Thus, although IL-6 and TNF, as well as IL-1, show common central effects such as fever and pituitary-adrenal activation, these effects may be independent of the activation of NE metabolism in the hypothalamus. PMID- 8242364 TI - Basal forebrain control of cortical cerebral blood flow is independent of local cortical neurons. AB - To determine whether intrinsic cortical neurons participate in mediating increases in cortical cerebral blood flow (CBF) in response to electrical stimulation of the basal forebrain (BF), cortical CBF was assessed by laser Doppler flowmetry in rats before and after unilaterally removing local cortical neurons with the excitotoxin ibotenic acid (IBO). On the first day of testing, CBF responses to right and left BF stimulation were nearly identical in right and left frontal cortices, corresponding to the frequency of stimulation, up to a maximum at 25 Hz (+180%). Subsequently, animals received a unilateral microinjection of IBO and a contralateral microinjection of phosphate-buffered saline (PBS) into the responsive cortical sites. After five days, responses in lesioned cortices were remarkably intact both in comparison to the contralateral PBS-injected site and to the same site tested prior to lesioning on day 1. IBO lesions of the response sites were histologically confirmed to extend through the entire depth of the frontal cortex and to encompass a large surface area (7.7 +/- 0.5 mm2). These results indicate that local cortical neurons are not critical to the mediation of increases in cortical CBF as elicited by BF stimulation. This study further supports the role of the BF as a distinct intracerebral neurogenic regulator of cortical CBF. PMID- 8242365 TI - Nociceptive C fibre input to the primary somatosensory cortex (SI). A field potential study in the rat. AB - In the present study, noxious thermal stimulation of the skin with short pulses of CO2-laser radiation was used to identify a cutaneous nociceptive C fibre input to SI and investigate the organization of this input in halothane-nitrous oxide anaesthetized rats. Noxious CO2-laser stimulation of the glabrous skin of the hindpaw consistently evoked late surface positive field potentials in SI (average onset latency 226 ms, peak latency 296 ms). It was demonstrated that these late potentials were evoked by an input from nociceptive C fibres, using a combination of latency measurements, anodal block of A fibre conduction and graded intensities of stimulation. Compared to the tactile evoked potentials in SI, the nociceptive C fibre evoked potentials were more widespread and exhibited a crude somatotopical organization. Intracortically, both tactile and nociceptive C fibre evoked potentials reversed polarity and exhibited a peak negativity in laminae III-IV. The nociceptive C fibre evoked potentials exhibited an additional peak negativity in laminae V-VI. The latter potential had a different time course as compared to the nociceptive C fibre potential evoked in laminae III-IV. In conclusion, an input from cutaneous nociceptive C fibres to SI was demonstrated for the first time in animal experiments. The input to SI from tactile receptors and cutaneous C nociceptors were differently organized. PMID- 8242366 TI - Multiple spinal pathways mediate cutaneous nociceptive C fibre input to the primary somatosensory cortex (SI) in the rat. AB - In the present study, partial lesions of the lower thoracic spinal cord in rats anaesthetized with halothane and nitrous oxide were made in order to elucidate which of the spinal funiculi mediate a nociceptive C fibre input to SI. Field potentials evoked by noxious CO2-laser stimulation were recorded in the left SI. Nociceptive C fibre input from the right hindpaw to SI was propagated by the dorsal funiculi (DF) and the left and right lateral funiculi (LLF and RLF, respectively). Nociceptive C fibre input from the left hindpaw was propagated by LLF and RLF, but not DF. Input from the hindpaws mediated by LLF and RLF caused widespread surface positive potentials throughout most of SI, although potentials in the hindlimb area tended to be larger than those in other areas of SI. Input from the right hindpaw mediated by DF caused surface positive potentials mainly in the hindlimb area of SI. Intracortically, the field potentials reversed polarity in the superficial laminae and had maximal negative amplitudes in laminae III-IV (input transferred by DF and LLF) and in laminae V-VI (input transferred by LLF and RLF). It is concluded that there are multiple spinal pathways which can transfer information from cutaneous nociceptive C fibres to SI in the rat. These ascending pathways seem to activate partly different thalamo cortical systems. PMID- 8242367 TI - Sensory responses to passive hindlimb joint rotation in the cerebellar cortex of the cat. AB - We recorded from Purkinje cells in the anterior and posterior lobes of the cerebellar vermis in anesthetized cats during passive flexion and extension of the ipsilateral hind foot. Nearly 60% of the 302 cells recorded in lobules I-IV anterior and VII-IX posterior responded to this stimulus. In lobule V, generally considered to be the forelimb projection area, about a third of the 38 cells tested responded to hind foot movement. Cells in the anterior lobe typically exhibited different poststimulus patterns in response to foot flexion and extension while posterior cells usually had the same response to these stimuli. We also noted that cells recorded in the medial vermis (within 2.7 mm of the midline) generally had longer response latencies than cells recorded more laterally (2.7 mm of the midline) generally had longer response latencies than cells recorded more laterally (2.7-4.5 mm) in both lobes. The results illustrate a diffuse proprioceptive input to the cerebellar vermis which has different features in various cerebellar areas. PMID- 8242368 TI - Surgical damage stimulates proliferation of dopamine uptake sites in normal mouse brain. AB - To test the hypothesis that proliferation of host dopaminergic tissue in response to injury plays an important role in the response to intrastriatal grafting, we transplanted autologous adrenal medullary to striatum in normal C57-black mice and compared this procedure with transplantation of non-dopaminergic tissue (frontal cortex) or a non-cellular matrix (Gelfoam). [3H]Mazindol autoradiography revealed that all three protocols resulted in a marked proliferation of dopamine uptake sites 10 months after transplantation. PMID- 8242369 TI - Acute withdrawal after repeated ethanol treatment reduces the number of spontaneously active dopaminergic neurons in the ventral tegmental area. AB - The effect of acute withdrawal, after repeated ethanol administration, on the electrophysiological activity of dopamine (DA) neurons in the ventral tegmental area was studied. Male rats received a 10-day treatment of ethanol at 4 g/kg, twice daily via intragastric intubation. In animals treated with ethanol, the number of spontaneously active DA neurons, determined with the cells-per-track population sampling technique, was significantly reduced (ethanol group = 0.70 +/ 0.10; control group = 1.49 +/- 0.18). I.V. apomorphine (20-64 micrograms/kg) reversed the reduced number of spontaneously active DA neurons (1.30 +/- 0.14) to near control levels. 'Silent' DA neurons could also be induced to fire by microiontophoretic application of GABA. These results suggest that the reduction in the number of spontaneously active DA neurons was due to the induction of tonic depolarization-inactivation on these neurons. PMID- 8242370 TI - Microiontophoresis of baclofen on membrane potential and input resistance in bulbar respiratory neurons in the cat. AB - Iontophoresis of baclofen produced hyperpolarization and a decrease in input resistance in 32 neurons and no effect in 24 neurons of the ventral respiratory group in cats. Iontophoresed phaclofen antagonized the effect of baclofen, but had negligible effects on periodic fluctuations in membrane potential and spike activity in these neurons. The hyperpolarizing effect of baclofen persisted after iontophoresis of tetrodotoxin, suggesting that baclofen acted directly at the postsynaptic gamma-aminobutyric acid (GABA)B receptors. PMID- 8242371 TI - Neurotrophic action of gliostatin on cocultured neurons with glial cells. AB - Gliostatin is a polypeptide factor (apparent M(r) = 100 k with a homodimeric structure comprising two 50 kDa subunits) acting on cortical neurons (neurotrophic action) as well as astrocytic cells (growth inhibition). Under the coculture system of cerebral cortical neurons and astrocytes from fetal rats (E15 or E16), the neurotrophic action of gliostatin was examined immunocytochemically. Immunostaining by an anti-neurofilament (NF) monoclonal antibody visualized a marked neurite-outgrowth and interconnecting bundles of neuritic processes induced by gliostatin in the coculture system. Neurons stimulated by gliostatin formed dense aggregates in clumps, while neurons in control coculture spread out. Gliostatin has also shown survival-promoting effects on neurons. Furthermore, it was shown that gliostatin induced the differentiation of protoplasmic astrocytes to fibrous astrocytes. These results further support our previous contention that gliostatin plays physiological roles on neuronal and glial development. PMID- 8242372 TI - Chromogranin A inhibits retinal dopamine release. AB - Chromogranin A (CGA), a peripherally active prohormone, is a soluble component in the secretory granules of many endocrine tissues and is cosecreted with their peptide or amine hormones. Using an antibody prepared against purified rat adrenal CGA, immunostaining was localized to the inner and outer plexiform layers of the rat retina and to selected ganglion cells. Exogenous CGA (purified from human adrenals) when applied to perfused rat retina potently inhibited the potassium-induced release of endogenous dopamine (DA). This action was dose dependent, with an IC50 of 3 nM; at 100 nM CGA retinal DA release was completely abolished. PMID- 8242373 TI - Basal and stress-induced corticosterone secretion is decreased by lesion of mesencephalic dopaminergic neurons. AB - There is evidence that certain psychopathological conditions are accompanied by a dysfunction in both the hypothalamo-pituitary-adrenal axis and dopaminergic systems, although the relationship between these two systems is as yet unclear. In the present study we investigated the effect of a specific lesion of dopamine mesencephalic neurons (Ventral Tegmental Area) on basal and stress-induced corticosterone secretion. Three weeks after injection of 6-OHDA, there was a depletion in dopamine in the frontal cortex and in the ventral and dorsal striatum, whereas norepinephrine and serotonin levels were unchanged. The dopamine-lesioned rats exhibited a lower basal and stress-induced corticosterone secretion than the sham-lesioned animals. The results indicate that the dopaminergic system may have a stimulatory influence on the hypothalamo-pituitary adrenal axis. PMID- 8242374 TI - Rats orally self-administer corticosterone. AB - Corticosterone, the major glucocorticoid in the rat, may modulate the reinforcing properties of addictive drugs as well as act as a positive reinforcer for intravenous self-administration. Since glucocorticoids are generally administered to humans via the oral route, we examined the ability of corticosterone to induce oral self-administration in the rat. In a first experiment, animals with free access to food could choose between a corticosterone solution and water. Three doses (25, 50 and 100 micrograms/ml) were tested. The group receiving the 100 micrograms/ml dose was also submitted to an extinction followed by a reversal test. In a second experiment, we examined whether the reinforcing properties of corticosterone could induce drinking independently of food intake. In the pre test phase rats had access to food only during a fixed period of the day (11.00 h to 14.00 h). Corticosterone solution (200 micrograms/ml) or tap water were available during this period, with free access to tap water for the rest of the day. During the test period, access to food was shifted forward in time, while the availability of the corticosterone solution remained the same. The first experiment showed that rats preferred a corticosterone solution to tap water, developing self-administration in a dose-dependent manner. This preference could be extinguished, but was regained during the reversal phase. In the second experiment, animals that had access to the corticosterone solution drank more than rats that had access to water in the absence of food. These results indicate that corticosterone has reinforcing properties after oral administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242375 TI - Peripheral projections of calretinin-immunoreactive primary sensory neurons in chick hindlimbs. AB - In chicken dorsal root ganglia, calretinin immunoreactivity is expressed by a subpopulation of large A-neurons, most of which co-express calbindin D-28k. The myelinated axons of these neurons selectively innervate all muscle spindles and most Herbst corpuscles associated to feathers in hindlimbs. It is suggested that the presence of calretinin in primary afferents may be correlated with the electrophysiological properties of rapidly adapting mechanoreceptors. PMID- 8242376 TI - MK-801 elevates the extracellular concentration of dopamine in the rat prefrontal cortex and increases the density of striatal dopamine D1 receptors. AB - In the present study we found that MK-801 (dizocilpine), given peripherally in doses (0.2 and 0.4 mg/kg) which evoked locomotor activation in rats, enhanced in a dose-dependent manner the extracellular concentration of dopamine (DA) in the rat prefrontal cortex (PFC). MK-801 used in similar range of doses (0.2, 0.4 mg/kg) failed to alter the DA content in superfusates of the rat striatum (STR). It was also found that single doses of MK-801 enhanced the density of D1 receptors, assessed by the [3H]SCH 23390 binding in the rat STR, but not in the limbic forebrain. An increase in the density of D1 receptors was observed at 24, but not 2, h after MK-801 administration. MK-801 failed to alter the density of D2 receptors in the STR and limbic forebrain. The available data indicate that MK 801 may enhance the dopaminergic neurotransmission by at least two separate mechanisms: a fast one, associated with the release of DA in PFC, and a slow one, resulting from the increase in the D1 receptor density. PMID- 8242377 TI - Glutamate receptor immunoreactivity in the rat suprachiasmatic nucleus. AB - Antibodies selective for the glutamate receptor subunits GluR1 and GluR2/3 were used to localize glutamate receptor immunoreactivity in the rat suprachiasmatic nuclei (SCN). These antisera identified two distinct cell populations within the SCN. Cells immunoreactive to GluR2/3 antiserum were located predominantly in the ventral SCN while antiserum selective for GluR1 stained a population located along the dorsal and lateral borders of the nucleus. There were no apparent day night differences in GluR immunoreactivity observed in the SCN. PMID- 8242378 TI - The dorsal cochlear nucleus of the adult lurcher mouse is specifically invaded by embryonic grafted Purkinje cells. AB - The fate of embryonic Purkinje cells grafted over the brainstem surface of the adult Lurcher mouse was analyzed using anti-calbindin (CaBP) immunocytochemistry. Purkinje cells are able to migrate specifically into the molecular layer of the host dorsal cochlear nucleus (DCoN) and develop dendritic trees that are practically isoplanar, suggesting synaptic interactions with the parallel fibres of the DCoN. These results provide a new argument in favour of the homology between the cerebellum and the DCoN. PMID- 8242379 TI - Ventral mesencephalic delta opioid receptors are involved in modulation of basal mesolimbic dopamine neurotransmission: an anatomical localization study. AB - Microdialysis was used to examine the anatomical localization of mesencephalic delta opioid receptors that participate in modulation of mesolimbic dopamine (DA) neurotransmission. Independent groups of rats were injected with DPDPE into the ventral tegmental area (VTA) or interpeduncular nucleus (IPN). Extracellular nucleus accumbens DA and DOPAC concentrations were elevated after DPDPE injections into either site, but injections into the VTA were effective at lower doses than were injections into the IPN. Thus, it appears that the DA-modulating actions of DPDPE are mediated in the VTA rather than the IPN. PMID- 8242380 TI - Alterations in the activity of adenylate cyclase and high affinity GTPase in Alzheimer's disease. AB - The aim of this study was to assess the effect of Alzheimer's disease has on the functional integrity of several signal transduction proteins. The relative levels of the G-protein alpha subunits Gs alpha-L, Gs alpha-S, Gi alpha-2 and G(o) alpha were measured by western blotting and found to be unchanged in membranes prepared from Alzheimer-diseased frontal cortex or hippocampus compared to control brains. However the activity of the G-protein associated enzyme, high affinity GTPase, was found to be reduced in the frontal cortex (reduced by 25%) and by a similar magnitude in the hippocampus (reduced by 27%) of Alzheimer subjects. The same membrane preparations were also assayed for the activity of adenylate cyclase. Basal enzyme activity was not significantly altered in Alzheimer diseased hippocampus, but was markedly reduced (by 45%) in the frontal cortex. The ability of fluoride and aluminium ions to stimulate adenylate cyclase was not significantly changed in either brain region. This suggests that G-proteins, especially Gs, are still able to interact with this enzyme. These results indicate that although the presence of Alzheimer's disease does not significantly alter G-protein levels, changes have taken place in the overall activity of these proteins. However this alteration does not affect their ability to stimulate adenylate cyclase activity. PMID- 8242381 TI - The interaction between central cholinergic and peripheral beta-adrenergic systems on radial maze performance in rats. AB - The interaction between cholinergic and beta-adrenergic systems on radial arm maze performance was investigated. Scopolamine significantly disrupted radial maze performance. Although neither propranolol (a centrally and peripherally acting beta-antagonist), nor atenolol (a peripherally acting beta 1-antagonist) affected the maze performance, both antagonists significantly augmented the scopolamine disruption. These results suggest the importance of the interaction between central cholinergic and peripheral beta-adrenergic systems on radial maze performance. PMID- 8242382 TI - Non-synaptic modulation of dorsal column conduction by endogenous GABA in neonatal rat spinal cord. AB - GABAA receptor activation can modulate axonal conduction in the isolated dorsal column of the neonatal rat spinal cord in vitro. However, it is not known whether axonal conduction in the dorsal column can be modulated by endogenous GABA in the developing spinal cord. We consequently compared the effects of GABA, a GABAA agonist, and a GABA uptake inhibitor on axonal conduction in the dorsal column of hemisected neonatal (0- to 9-day-old) rat spinal cords in vitro. Extracellular compound action potentials evoked by supramaximal stimuli were recorded at two points with glass microelectrodes. GABA (10(-4) to 10(-3) M) reversibly decreased the compound action potential amplitude and the population conduction velocity. At 10(-4) M, compound action potential amplitudes fell by 45.0 +/- 6.5% of control while the conduction velocity slowed by 11.8 +/- 4.3% (n = 5). The GABAA receptor agonist, isoguvacine, mimicked the effects of GABA on the dorsal column compound action potential. In contrast, while GABA at 10(-5) M decreased the amplitude by 7.7 +/- 3.1%, it increased conduction velocity by 9.7 +/- 1.3% (n = 5). The GABA uptake inhibitor, nipecotic acid (10(-3) M), consistently decreased the compound action potential amplitude by 17.7 +/- 6.5% (n = 6) but the conduction velocity slowed in four out of six preparations. In two instances, nipecotic acid decreased the amplitude and increased the conduction velocity. The effects of nipecotic acid on the dorsal column compound action potential were blocked in the presence of the GABAA antagonist bicuculline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242383 TI - Depletion of olfactory bulb norepinephrine by 6-OHDA disrupts chemical cue but not social recognition responses in male rats. AB - In the present experiment, 6-OHDA was infused directly into the olfactory bulb (OB) to produce a localized neurotoxic lesion. Habituation/dishabituation behavioral tests were then conducted to measure recognition responses to chemical cues (urine as a stimulus) and to social stimuli (ovariectomized rat as a stimulus). Infusion of 6-OHDA resulted in a near complete depletion of OB norepinephrine (NE), whereas it had little effect (15% reduction) on OB dopamine (DA) contents. Nor were any significant effects on hypothalamic, hippocampal, olfactory tubercle, and corpus striatal NE and DA contents observed. Behaviorally, dishabituation responses to chemical cues were greatly impaired, however, there was relatively little effect on social behavior dishabituation responses. These results demonstrate that 6-OHDA can be used to produce a near complete but localized depletion of OB-NE. This treatment impairs dishabituation responses to chemical cues but not social stimuli indicating that OB-NE appears necessary for processing of chemical cue, but not social memory recognition process. PMID- 8242384 TI - Circadian phase-shifts induced by chlordiazepoxide without increased locomotor activity. AB - Phase advances of circadian locomotor rhythms in response to the benzodiazepine triazolam (TRZ) administered at circadian time 6 appear to be mediated by locomotor activity. The possibility that increased activity also mediates shifts induced by the benzodiazepine chlordiazepoxide (CDZ) was investigated. Hamsters injected with CDZ 7 h before dark onset exhibited phase advances despite being confined to their nest boxes for 3 h after receiving the drug. These shifts did not differ significantly from advances seen in hamsters which were allowed access to the rest of the cage after injections. In other experiments, the behavioral effects of both CDZ and TRZ were examined during the 5 h after injection. There was significantly more motor activity observed after injection of TRZ than CDZ. These results suggest that phase-shifting effects of some benzodiazepines can occur without inducing activity. PMID- 8242385 TI - Distribution of trkB tyrosine kinase immunoreactivity in the rat central nervous system. AB - Recent evidence suggests that trkB tyrosine kinase is a high affinity receptor for brain-derived neurotrophic factor (BDNF). BDNF can act as a survival factor for several neuronal subgroups and its mRNA is distributed widely throughout the central nervous system. However, the functional targets of BDNF are poorly defined. We have used immunochemical and immunohistochemical techniques to determine the regional distribution and cellular localization of trkB tyrosine kinase-like immunoreactivity. The staining pattern indicates that the trkB-like antigen is widely distributed and present within both glia and neurons. Astrocytes were the most intensively labelled but many neuronal populations were also stained. In some regions including brain stem, spinal cord, hippocampus and diagonal band of Broca, neurons were stained at varying intensities. In other areas such as the cortex of the forebrain and amygdaloid nucleus, the stain was intense but diffuse, preventing positive identification of the cell types involved. Immunoblot results indicated two separate protein bands in all brain and spinal cord regions examined, of molecular weights 145 and 85 kDa, respectively. These findings aid the definition of neuronal and glial subpopulations of the central nervous system that may utilize BDNF. PMID- 8242386 TI - Intraspinal modulation of neuronal responses to uterine and cervix stimulation in rat L1 and L6 dorsal horn. AB - This two-part study examined intraspinal processing of input from the uterus, cervix and skin within the caudal spinal cord of virgin rats in estrus. The first part examined the effects of removing either pelvic or hypogastric nerve input (by appropriate dorsal rhizotomies) on neuronal responses to stimulation of uterus, cervix and skin in either the L1 or L6 dorsal horn of decerebrate, spinalized unanesthetized rats. Bilateral section of the T13-L2 roots (through which most hypogastric afferents travel) eliminated almost all responses to uterine distension in L1 and L6. Such rhizotomies also decreased the inhibitory effects of cervix input on neurons in L6. Bilateral section of the L6-S2 roots (through which most pelvic afferents travel) had no effect on responses to uterus, but decreased the inhibitory effects of cervix input on neurons in L1. Sections of both sets of roots eliminated responses to cervix stimulation in L6. The second part examined neuronal responses in T13/L1 of urethane-anesthetized rats before and after a T10 spinal transection. Transection increased the probability of observing neurons excited by uterine stimulation. Transection also increased excitatory responses and decreased inhibitory responses to cervix stimulation. These results confirm previous findings that input from the uterus to the spinal cord is mainly by way of the hypogastric nerve, while that from the cervix is by way of both the pelvic and hypogastric nerves. The results also demonstrate that descending influences in the estrous rat mainly increase the ability of cervix stimulation to inhibit spinal neuronal activity and reduce the effectiveness of uterine stimulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242387 TI - Blockade of 45Ca2+ influx through the N-methyl-D-aspartate receptor ion channel by the non-psychoactive cannabinoid HU-211. AB - The effects of the synthetic non-psychoactive cannabinoid (+)-(3S,4S)-7-hydroxy delta 6-tetrahydrocannabinol 1,1-dimethylheptyl (HU-211) on the activity of the N methyl-D-aspartate (NMDA) receptor/ion channel were examined. HU-211 non competitively blocks the increase in binding of [3H]N-[1-(2-thienyl) cyclohexyl]piperidine ([3H]TCP) induced by the polyamines spermine and spermidine or by glutamate and glycine. HU-211 does not, however, affect the direct binding of [3H]glycine and [3H]glutamate to their binding sites on the NMDA receptor, which suggests that the effects of HU-211 are not mediated via the binding sites of glutamate-, glycine- and phencyclidine-like drugs or of polyamines. HU-211 can also block 45Ca2+ uptake through the NMDA-receptor/ion channel in primary cell cultures of rat forebrain. All of the above inhibitory effects of HU-211 on the NMDA-receptor/ion channel activity are stereospecific, since the (-)(3R,4R) enantiomer (HU-210) is ineffective. PMID- 8242388 TI - Age-related changes in receptor-mediated and depolarization-induced phosphatidylinositol turnover in mouse brain. AB - The effect of aging on receptor- and G-protein-activated and on depolarization induced phosphoinositide (PI) hydrolysis was examined in mechanically dissociated neurons from female NMRI mice. Additionally, age-dependent changes in Ca2+ homeostasis, i.e. changes in basal intracellular calcium ([Ca2+]i) and in depolarization-induced rise in [Ca2+]i were investigated. No age-related differences in PI hydrolysis were found after stimulation of muscarinic cholinergic, alpha 1, serotonin and quisqualate receptors coupled to the phosphoinositide-phospholipase C (PI-PLC) system. PI hydrolysis following stimulation with AMPA ((RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) revealed a significantly increased response in aged animals. Activation of G-proteins with NaF also induced a higher inositol monophosphate (InsP1) accumulation in aged mice. Moreover, InsP1 accumulation due to PLC activation by increased [Ca2+]i after depolarization with KCl was significantly increased in neurons from aged animals. Investigations about age-related changes in Ca2+ homeostasis revealed lower basal [Ca2+]i and lower rise in [Ca2+]i after depolarization with KCl. The data indicate that receptor-mediated and depolarization-induced PI hydrolysis are differentially affected by aging. Decreased availability of [Ca2+]i in aged animals may enhance the sensitivity of Ca(2+)-activated mechanisms. This may explain increased KCl- and AMPA-induced InsP1 accumulation whereas receptor-coupled PLC activation is less affected. PMID- 8242389 TI - Localization and characterization of vasopressin binding sites in the rat brain using an iodinated linear AVP antagonist. AB - The binding characteristics and central distribution of 125I-Linear AVP antagonist, a new ligand for vasopressin binding sites, are described in the following studies. Saturation studies performed on rat brain septal membranes demonstrated that 125I-Linear AVP antagonist binds to a single class of sites with high affinity (55 pM) and limited capacity (88 fmol/mg protein). In autoradiographic studies, 125I-Linear AVP antagonist labeled brain areas known to contain vasopressin receptors without binding to neurophysins. 125I-Linear AVP antagonist also labeled sites in cortex, hypothalamus, ventral tegmental area and substantia nigra. In competition studies, 125I-Linear AVP antagonist binding was most readily blocked by AVP and a selective V1a agonist. Oxytocin and a selective V2 ligand were effective only in micromolar concentrations. A selective oxytocin agonist was virtually ineffective in blocking 125I-Linear AVP antagonist binding. In regions that contain a high density of oxytocin binding sites, however, oxytocin-displaceable binding was observed. In agreement with studies on peripheral tissues, the binding profile generated from these studies indicates that 125I-Linear AVP antagonist binds to vasopressin receptors of the V1a subtype. These results suggest that 125I-Linear AVP antagonist is a valuable ligand for the study of central AVP receptors. PMID- 8242390 TI - Effect of thyrotropin releasing hormone on U-50,488H-induced pharmacological responses in mice. AB - The effect of thyrotropin releasing hormone (TRH) administered either subcutaneously (s.c.) or intracerebroventricularly (i.c.v.) on the analgesic and hypothermic actions of U-50,488H, a highly selective kappa-opiate agonist, was determined in male Swiss Webster mice. Intraperitoneal administration of U 50,488H (8-32 mg/kg) produced a dose-dependent analgesia as assessed by the tail flick test. Similarly, U-50,488H also produced a dose-dependent hypothermia in mice. TRH was administered s.c. 15 min or i.c.v. 5 min prior to U-50,488H injection. TRH (1,3 and 10 mg/kg, s.c.) dose-dependently attenuated the analgesic effect of U-50,488H (32 mg/kg), whereas TRH at these doses displayed almost complete blockade of the hypothermic effect of U-50,488H. Similarly, TRH (0.03, 0.3 and 1 microgram/mouse; i.c.v.) dose-dependently attenuated the analgesic and hypothermic actions of U-50,488H, indicating the central component in the action of TRH. TRH alone in doses used showed no change in either basal tail-flick latency or body temperature, demonstrating the lack of effect of this drug alone on pain and temperature responsiveness. Studies have shown that TRH does not modify morphine or beta-endorphin-induced analgesia in animals nor does it affect the binding of mu-opiate agonist or antagonist to brain membranes. Previous studies from this laboratory have indicated that kappa-opiates but not mu-opiates inhibit the binding of [3H][3-MeHis2]-TRH to brain membranes. The present studies clearly show that TRH modulates the pharmacological actions mediated by kappa opiate agonists in mice. Thus, these studies provide further in vivo evidence for an acute interaction between TRH and kappa-opiates. PMID- 8242391 TI - Immunocytochemical distribution of peptidergic and cholinergic fibers in the human amygdala: their depletion in Alzheimer's disease and morphologic alteration in non-demented elderly with numerous senile plaques. AB - As part of an ongoing investigation devoted to understanding the pathogenesis of senile plaques, we employed histochemical and immunocytochemical techniques to examine the distribution and cytologic features of acetylcholinesterase (AChE), choline acetyltransferase (ChAT), somatostatin (SOM), neurotensin (NT) and substance P (SP) containing fibers and neurons within the amygdala of: (1) patients with Alzheimer's disease (AD); (2) age-matched non-demented controls (NC); and (3) a group of non-demented cases, who upon postmortem neuropathologic examination exhibited sufficient numbers of senile plaques to be classified as AD. This latter group was referred to as high plaque non-demented (HPND). For every case, the distribution of immunolabeled fibers and neurons were determined for each transmitter throughout the various subnuclei of the amygdala. In addition, in the AD and HPND cases the topographic distribution of senile plaques was determined throughout the amygdala using thioflavine-S and Bielschowsky silver methods. In the amygdala, the distribution and density of senile plaques were not bound by conventional cytoarchitectural groupings but rather were most dense in the ventromedial regions of the amygdala with decreasing density in dorsal and lateral directions. Importantly, the density and distribution of senile plaques failed to correlate with the normal topography and/or density of the various peptidergic or cholinergic fibers within the amygdala. The finding that plaques do not correlate with the topographic distribution of any specific transmitter system suggests that plaques likely do not arise from the degeneration of a single neurotransmitter system (i.e., the cholinergic system). However, the finding that in AD a transmitter is most markedly depleted in regions of greatest plaque density, suggests certain constituents of the plaque (e.g. beta-amyloid) may be contributing to the degeneration of local fibers. The extent to which a transmitter was depleted in AD patients varied considerably among those four investigated with the cholinergic and NT systems displaying the most dramatic reductions, followed by SP and SOM. Despite these differential reductions in fiber density, all four neurotransmitters were found localized within dystrophic neurites and in most instances these dystrophic neurites were associated with thioflavine-positive senile plaques. In contrast to the AD cases, the HPND cases were characterized by no significant reductions in immunolabeled fibers, although immunostained dystrophic neurites were very prevalent in the HPND cases. These data suggest that dystrophic neurites occur very early in the disease process and likely precede the actual loss of fibers when or if it occurs.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8242392 TI - Increases of CCK mRNA and peptide in different brain areas following acute and chronic administration of morphine. AB - The present study examined whether either acute or chronic administration of morphine resulted in changes in the content of CCK mRNA and CCK immunoactive peptide in selective areas of the rat brain and spinal cord. Two hours after a single injection of morphine (10 mg/kg, s.c.), CCK mRNA significantly increased in the hypothalamus (0.8-fold) and spinal cord (2-fold) relative to the CCK mRNA content in saline-injected controls. No significant differences in CCK mRNA were observed in the frontal cortex, hippocampus, midbrain or brainstem. There were no significant alterations in CCK immunoreactivity in any brain regions and spinal cord after the acute treatment with morphine. Upon repeated morphine administration, the content of CCK mRNA in both the hypothalamus and the spinal cord was further elevated by at least 3-fold. A significant increase of CCK mRNA content in brain stem (2.8-fold) was also observed following chronic morphine administration. In contrast to the acute exposure to morphine, chronic administration resulted in significant increases in CCK immunoactive peptide in hypothalamus (2.6-fold), spinal cord (2.1-fold) and brainstem (1.6-fold), but not in the other brain areas. These results demonstrate that morphine, especially following repeated administrations, stimulates endogenous CCK biosynthesis in selective brain regions. PMID- 8242393 TI - Immunocytochemical localization of nicotinic acetylcholine receptor in rat hypothalamus. AB - Immunocytochemical localization of neuronal nicotinic acetylcholine receptor (nAChR) was examined in rat hypothalamus. Monoclonal antibody against alpha 4 ACh binding subunits of nAChR was used in the avidin-biotin-peroxidase complex (ABC) immunocytochemical method at both the light and electron microscopic levels. By light microscopy nAChR-like immunoreactivity was found in many neuronal cell bodies and their fibers in the paraventricular nucleus (PVN) and in many axons and axon terminals in the median eminence (ME). The immunoreactivity of nAChR was the most intense in the ME. By electron microscopy immunoreaction products occurred on the rough endoplasmic reticulum, nuclear envelope, cytoplasmic matrices and postsynaptic densities of synaptic junctions in some neurons in the parvocellular part of the PVN. In the external layer of the ME, nAChR-like immunoreactivity was found over the entire plasma membranes of many axon terminals. Involvement of nAChRs in the release of neurotransmitters and neuropeptides both in the PVN and the ME is discussed. PMID- 8242394 TI - Daily changes in the release of acetylcholine from rat primary somatosensory cortex. AB - Using microdialysis, acetylcholine (ACh) release was measured in the somatosensory cortex of 14 rats over a 24-h period. The release of ACh was 0.195 pmol/min during the day and 0.344 pmol/min at night. The length of exposed dialysis membrane within the cortex was an important source of variability in the absolute amounts of ACh collected. Even after rejecting some cases where the membrane contacted only the superficial cortical layers, this factor accounted for 25% of the variation of absolute amounts collected in different animals. After correcting for the length of exposed membrane, the release of ACh was shown to increase 52% at night during the time when the animals were awake, feeding and grooming. Variability in the measures of ACh release obtained during periods of activity was greater than its variability during periods of inactivity. These data were interpreted in the context of several hypothesized roles for ACh in sensory cortex. PMID- 8242395 TI - Dopamine induces sign reversal at mixed chemical-electrical synapses. AB - A mixed chemical/electrical synapse can generate variable output when the strength of each synaptic component is modulated. At mixed synapses of the lobster pyloric network, the chemical component is inhibitory. Without neuromodulation, the chemical component is weak or absent and the electrical component often dominates. Dopamine reverses the sign of these mixed synaptic interactions by a reduction in the strength of electrical coupling and an enhancement of chemical inhibition, including activation of silent chemical synapses. Sign reversal at mixed synapses by neuromodulators may contribute to functional rewiring of neural networks. PMID- 8242396 TI - Peptide-induced grooming behavior and caudate nucleus dopamine release. AB - We simultaneously measured the display of grooming behavior and, by monitoring the extracellular dopamine concentration via transversal microdialysis, the release of dopamine in the caudate nucleus in freely moving rats after i.c.v. administration of 1 micrograms adrenocorticotropic hormone-(1-24) (ACTH-(1-24)). During a period of 1 h after administration of the peptide, the incidence of excessive grooming behavior was increased. Concomitantly, the concentration of dopamine in the caudate nucleus dialysates was significantly increased (maximal effect 151% of basal release) whereas that of its metabolite DOPAC was unchanged. The potent alpha-melanocyte stimulating hormone (alpha-MSH) receptor agonist, [Nle4,D-Phe7]alpha-MSH, induced grooming behavior and stimulated caudate nucleus dopamine release (maximal effect 148% of basal release) whereas ACTH-(7-16)-NH2 did neither induce grooming behavior nor cause an increase in caudate nucleus dopamine release. Single-dose tolerance was observed for ACTH-induced grooming but not for ACTH-induced dopamine release. These data are in support of the proposed involvement of brain dopamine systems in grooming behavior of the rat but at the same time suggest that the effect of ACTH/MSH-like peptides on dopaminergic transmission in the caudate nucleus is proximal to the final neural pathway involved in ACTH-induced grooming behavior. PMID- 8242397 TI - Effects of nicotine on cortical high voltage spindles in rats. AB - Cholinergic systems have been shown to modulate 6-10 Hz immobility-related cortical spike wave discharges (high voltage spindles-HVS) in rats. This study reports that activation of central nicotinic receptors inhibits HVS identified from cortical EEG recordings. Nicotine (0.19-1.9 mumol/kg i.p.) significantly reduced the summed duration of HVS bursts during 20 min of waking immobility. The nicotinic antagonist mecamylamine (5.0 mumol/kg i.p.) blocked the effect of nicotine (0.62 mumol/kg i.p.) without itself significantly affecting HVS. At higher doses, mecamylamine (15.0 and 25.0 mumol/kg i.p.) increased HVS activity. Dimethylphenylpiperazinium (0.62-6.2 mumol/kg i.p.), a nicotinic agonist which does not cross the blood-brain barrier, did not affect HVS, consistent with the idea that the effect of nicotine on HVS is due to an action in the central nervous system. Cotinine, the major metabolite of nicotine, did not affect HVS at doses similar to or higher than those tested for nicotine. Cotinine also did not block the effect of nicotine, indicating that this metabolite does not interfere with the modulatory effect of nicotine on HVS. These results suggest a role for nicotinic regulation of the neuronal substrates involved in the generation of HVS. PMID- 8242398 TI - MK-801, but not drugs acting at strychnine-insensitive glycine receptors, attenuate methamphetamine nigrostriatal toxicity. AB - Repeated administration of methamphetamine (METH) results in damage to nigrostriatal dopaminergic neurons. Both competitive N-methyl-D-aspartate (NMDA) receptor antagonists and use-dependent cation channel blockers attenuate METH induced damage. The objectives of the present study were to examine whether comparable reductions in METH-induced damage could be obtained by compounds acting at strychnine-insensitive glycine receptors on the NMDA receptor complex. Four injections of METH (5 mg/kg i.p.) resulted in a approximately 70.9% depletion of striatal dopamine (DA) and approximately 62.7% depletion of dihydroxyphenylacetic acid (DOPAC) content, respectively. A significant protection against METH-induced DA and DOPAC depletion was afforded by the use dependent channel blocker, MK-801. The competitive glycine antagonist 7 chlorokynurenic acid (7-Cl-KA), the low efficacy glycine partial agonist (+)-3 amino-1-hydroxy-2-pyrrolidone ((+)-HA-966), and the high efficacy partial glycine agonist 1-aminocyclopropane-carboxylic acid (ACPC) were ineffective against METH induced toxicity despite their abilities to attenuate glutamate-induced neurotoxicity under both in vivo and in vitro conditions. These results indicate that glycinergic ligands do not possess the same broad neuroprotective spectrum as other classes of NMDA antagonists. PMID- 8242399 TI - Photochemically induced focal cerebral ischemia in rat: time dependent and global increase in expression of basic fibroblast growth factor mRNA. AB - Induction of basic fibroblast growth factor (bFGF) mRNA expression was studied in a Rose bengal induced focal cerebral ischemia during a time course of 2, 4, 24, 72 h and 7 days. Focal cerebral ischemia induced by Rose bengal resulted in a global upregulation in bFGF gene expression at the 24 h time-interval. This upregulation in bFGF gene expression was due to an upregulation in glial bFGF expression in most of the areas studied as seen by means of non-radioactive in situ hybridization in combination with immunocytochemistry for glial fibrillary acidic protein. However, in the piriform cortex a putative neuronal upregulation of bFGF could be detected by combination of non-radioactive in situ hybridization, immunohistochemistry for glial fibrillary acidic protein and nuclear staining with Neutral red. Semiquantitative data concerning bFGF mRNA expression were obtained by use of computer-assisted microdensitometry and revealed substantial increases in bFGF mRNA expression in the cingulate cortex, the neostriatum, a 1 mm marginal zone close to the external capsule and the olfactory tubercle at bregma levels 1 to 2 mm rostral to the lesion. No changes in bFGF gene expression were seen in field CA1 of Ammon's horn on the lesioned side and in dentate gyrus at bregma levels between -2.12 to -3.30 mm. We observed significant changes in bFGF upregulation in the caudate putamen, the piriform cortex and the amygdaloid region and the frontoparietal cortex at bregma levels 2.12 to -3.30 mm. These data indicate that photochemically induced focal cerebral ischemia leads to an early and global response in bFGF gene expression, which is due to an upregulation mainly in astrocytes. The observed widespread upregulation of the bFGF gene transcription rostral and caudal to the lesion is suggested to be due in part to neuronal glutaminergic connections between the areas investigated and in part due to increases in extracellular fluid signals (volume transmission). PMID- 8242400 TI - The retrograde tracer Fluoro-Gold interferes with the infectivity of herpes simplex virus. AB - Fluoro-Gold has been used previously to identify those trigeminal ganglion cells that innervate the central cornea. To examine the effects of Fluoro-Gold treatment on infection and spread of HSV in vivo, we measured the number of plaque forming units recovered from trigeminal ganglia 3 or 5 days after corneal scratch and inoculation with Fluoro-Gold and HSV. Treatment with Fluoro-Gold reduced the amount of virus recovered after retrograde transport 63% at 3 days and 28% at 5 days after inoculation. When we examined trigeminal ganglion sections from animals treated with HSV and Fluoro-Gold, we found the number of neurons double labeled with antibodies that recognize HSV and Fluoro-Gold was only 13% of all Fluoro-Gold labeled neurons. This was significantly fewer cells that we had anticipated, on the basis of double labeling experiments with wheat germ agglutinin combined with Fluoro-Gold. The effects of varying doses of the retrograde tracer, Fluoro-Gold on Herpes simplex virus (type 1) (HSV) infectivity were also assayed in vitro using a standard viral plaque assay. At 1 x 10(-3) mg/ml Fluoro-Gold there was no effect on the number of plaque forming units. At 5 x 10(-1) mg/ml the number of plaques was reduced about 67%. We conclude that Fluoro-Gold interferes with productive HSV infection in vivo and in vitro after retrograde transport of HSV by neurons. PMID- 8242401 TI - Extracellular recordings in the colchicine-lesioned rat dentate gyrus following transplants of fetal dentate gyrus and CA1 hippocampal subfield tissue. AB - Grafts of fetal dentate gyrus (DG) and CA1 hippocampal subfield tissue were extruded into the dentate gyri of adult male Sprague-Dawley rats, 7-10 days after lesioning the granule cells with colchicine (0.06 microliter of 7 mg/ml solution at each of 5 sites/hippocampus). Graft area-host and host-graft area connectivities were investigated 4-6 months post-transplantation by recoding extracellular evoked response in hippocampal slice preparations. Following stimulation of the host mid-molecular layer, evoked field potential responses, showing considerable variation, were recorded in both types of graft. Evoked responses in the lesioned DG without grafts were recorded in very few slices. Stimulation of the area of DG tissue grafts occasionally evoked responses in the host CA3/CA4 and there was no evidence for CA1 graft area-CA3/CA4 connectivity; stimulation of DG and CA1 graft areas occasionally evoked responses in the host CA1. Responses in the area of both DG and CA1 grafts supported short-term potentiation following stimulation of the host mid-molecular layer but only DG graft areas supported long-term potentiation of the population spike amplitude. In the area of both types of transplant a tonic bicuculline-sensitive inhibition was present and paired-pulse stimulation paradigms provided some evidence for inhibition. It is possible that responses recorded within the area of grafted tissue to stimulation of the host are attributable to host-graft connectivity and similarly, responses recorded in the host to stimulation of the area of the graft may be attributable to graft-host connectivity. Only DG graft areas received host inputs which were capable of sustaining a long-term potentiation and establishing efferent contacts with the host CA3/CA4 subfield, suggesting that these would be more likely than CA1 grafts to reinstate normal functional circuitry. PMID- 8242402 TI - Corticosterone is a preferable ligand for measuring rat brain corticosteroid receptors: competition by RU 28362 and RU 26752 for dexamethasone binding in rat hippocampal cytosol. AB - It is unclear whether in vitro corticosteroid receptor binding assays have used inappropriately high concentrations of synthetic corticosteroid competitors, thereby potentially introducing error into estimates of type I (mineralocorticoid) and type II (glucocorticoid) receptor binding. To determine more accurately the concentration of blockers necessary to discriminate between these two sites, we have derived Ki values for the competition of dexamethasone, RU 28362 and RU 26752 for [3H]corticosterone and [3H]dexamethasone binding in rat hippocampus. Non-specific binding of both radioligands was defined with unlabeled dexamethasone to exclude transcortin. The type II agonist RU 28362 competed for only a portion of [3H]corticosterone binding, exhibiting a Ki of 0.5 nM for this binding. In contrast, RU 28362 fully competed all binding of a saturating concentration of [3H]dexamethasone, even though [3H]dexamethasone also recognized type I receptors, defined as specific [3H]corticosterone binding in the presence of 80 nM RU 28362. RU 28362 competition for [3H]dexamethasone binding exhibited characteristics of a 2-site interaction, with Kis of 0.3 and 194 nM. The type I receptor antagonist RU 26752 competed less effectively for [3H]corticosterone and [3H]dexamethasone binding, but nonetheless competed fully within a 1000-fold concentration range. Even at a level less than 125 x its Ki for type I binding, RU 26752 still inhibited virtually all type II receptor binding by [3H]corticosterone. We conclude that type I and II receptors in rat brain are best distinguished using [3H]corticosterone as the labelling ligand, with cold RU 28362 and dexamethasone to eliminate binding to type II and transcortin sites, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242403 TI - Vibration on poultry transporters. AB - 1. This research established the magnitudes and frequencies of vibration acceleration on two typical broiler bird transporters, one with leaf and the other with air suspension. 2. Vibration measurements were made using triaxial accelerometers mounted on the chassis and in the bird containers, with vehicles both loaded and unloaded. 3. In the vertical axis the fundamental vehicle frequency is between 1 and 2 Hz with a secondary peak at 10 Hz. 4. In the lateral axis with air suspension, the maximum level of vibration on the chassis occurred at 12 to 18 Hz; this was less evident with leaf suspension. 5. The magnitudes of vibration acceleration occurring in the bird containers would be fairly uncomfortable for the seated human. Frequency weightings and response to different axes and magnitudes of vibration must be established before the response of birds can be predicted. PMID- 8242404 TI - Are genetically lean broilers more resistant to hot climate? AB - 1. Genetically lean (LL) or fat (FL) male chickens were exposed to either high (32 degrees C) or control (22 degrees C) ambient temperature up to 9 weeks of age. They were fed on one of two isoenergetic diets differing in protein content: 190 or 230 g/kg. 2. At 22 degrees C, weight gain of LL broilers was the same as in FL chickens, but at the high temperature LL birds grew to a greater weight than FL ones. 3. Food conversion efficiency was not affected by ambient temperature in LL chickens but was depressed in FL ones at 32 degrees C. 4. Increasing dietary protein content did not alleviate heat-induced growth depression irrespective of the genotype. 5. Gross protein efficiency was higher in LL chickens and was less depressed at 32 degrees C than in FL birds. 6. Fat deposition decreased with increasing protein concentration at normal temperature in both genotypes; at high temperature, high protein content enhanced fatness, particularly in LL chickens. 7. Thus, genetically lean broilers demonstrated a greater resistance to hot conditions: this was indicated by enhanced weight gain and improved food and protein conversion efficiencies. PMID- 8242405 TI - Effect of spray sanitising on hatching egg cuticle efficacy and hatchability. AB - 1. Eggs from a broiler breeder flock were sanitised using a continuous spray sanitising machine. The effect of this treatment on the ability of the cuticle to resist water and bacterial penetration was investigated throughout the production cycle. 2. The volume of water taken up by eggs that had been sanitised was significantly (P < 0.001) greater than that taken up by the control eggs. The volume of water taken up by the sanitised eggs increased as the flock age increased, whereas water uptake was maximal for the control eggs at 42 weeks of age. 3. There was no correlation between water uptake values and shell thickness or egg weight for either the sanitised or control eggs. 4. Sanitised eggs tended to dominate the "poor cuticle quality" category at all flock ages, when a subjective assessment of cuticle quality was made using the stain Edicol Supra Pea Green. 5. Egg sanitising did not affect shell reflectance values, shell thickness or the percentage of cracked shells. 6. When flock age reached 49 weeks of age, the sanitising process significantly (P < 0.001) reduced hatchability. PMID- 8242406 TI - Diurnal-nocturnal changes in food intake, gut storage of ingesta, food transit time and metabolism in growing broiler chickens: a model for temporal control of energy balance. AB - 1. Diurnal-nocturnal changes in food intake, gut storage of ingesta, food transit time and heat production were studied in male broiler chickens reared under a 14L:10D lighting schedule (lights on from 06.00 to 20.00 h). 2. Food consumption during the scotophase was negligible. Peak food consumption during the photoperiod occurred at the beginning of the photoperiod and in the late afternoon. 3. During the photoperiod, the crop and proventriculus/gizzard contained only small quantities of ingesta. However, at the beginning of the scotoperiod, dried ingesta content of crop and proventriculus/gizzard increased by 10.5- and 2.76-fold respectively. This increase was followed by a gradual decrease towards the end of the scotoperiod. 4. Food transit time during the scotoperiod was significantly longer than that during the photoperiod. 5. The daily pattern of heat production closely followed the 14L:10D lighting schedule. Total heat production during darkness averaged 53% of total heat production during the photoperiod. 6. It was estimated that the storage of energy (as ingesta) in the crop and proventriculus/gizzard, followed by its gradual release and the increased food transit time during the night, contributed 75.5% of nocturnal energy needs. It must be recognised that these mechanisms play a major role in the energy balance of the growing chicken during periods without food intake. PMID- 8242407 TI - Effect of dietary protein and urea on in vitro caecal ammonia production from urea and uric acid in cockerels. AB - 1. Single Comb White Leghorn adult cockerels were fed on 50 g/kg protein diet, 200 g/kg protein diet or 50 g/kg protein diet plus urea and in vitro ammoniagenesis from urea and uric acid in the caeca was determined. 2. Four-fold protein intake caused about 4.6-fold increase in caecal ammonia production from urea (P < 0.05), and tended to increase it from uric acid as compared with 50 g/kg protein-fed birds. 3. Dietary urea significantly increased caecal ammonia production from urea and uric acid by about 2 and 3 times as much as those of control birds, respectively (P < 0.05). 4. It is concluded that increased protein intake and the feeding of urea are able to induce ammoniagenesis from urea and uric acid in the caeca of fowls. PMID- 8242408 TI - Food intake adjustments of chicks: short term reactions of genetic stocks to deficiencies in lysine, methionine or tryptophan. AB - 1. Growth, food intake, and food preference were measured in two experimental lines (White Rocks and White Leghorns) of chickens fed diets varying in content of lysine, methionine, or tryptophan. Diet A was balanced in all amino acids, while the remaining 6 diets were moderately (approximately 30%) or slightly (approximately 15%) deficient in either lysine, methionine, or tryptophan relative to NRC recommendations for broiler diets. 2. Body weight and food utilisation efficiency of White Leghorns at 10 d of age were not affected by diet, whereas the lower concentrations of lysine and methionine depressed these traits in White Rocks. 3. Tryptophan deficiency reduced food intake of White Leghorns, but not White Rocks. 4. With choice feeding of balanced and deficient diets from day 10 to 16, preferences between the diets were minimal for White Leghorns, whereas for White Rocks there was an almost immediate preference for the balanced diet, followed by lack of preference or preference for the deficient diet. 5. Apparently for some genetic stocks there is rapid adjustment to deficiencies in these amino acids with overconsumption, followed by compensation for preference of the balanced diet. The response, however, will vary depending on the deficient amino acid. PMID- 8242409 TI - Food intake adjustments of chicks: short term reactions to deficiencies in lysine, methionine and tryptophan. AB - 1. Two experiments were conducted to compare food intake responses of broiler chicks fed diets varying in lysine, methionine, and tryptophan. Diet D was formulated to create simultaneous deficiencies of lysine, methionine, and tryptophan. Diet A matched National Research Council (1984) recommendations for broilers, and diets B and C were, respectively, 2:1 and 1:2 mixes of diets A and D. 2. Short-term food intake can provide information on the sequences of adaptation of chicks to a diet deficient in essential amino acids. 3. Chicks consumed 26% less of diet D than A during the first 24 h posthatch. When chicks fed diet A or D to 7 d of age were then fed one of 4 diets singly, within 24 h intake was lowest for chicks fed diet D. Within 48 h, food intake of diet C was more than that of diet D and less than that of diet A, while for diet B intake was more than of diet D but not different from diet A. 4. In the second experiment, chicks were fed diet A to 8 d and then diets A or D alone or given a choice of diets A and D from 8 to 20 d of age. Within 4 to 8 h, food intake of chicks fed diet D alone decreased markedly followed by partial recovery within 24 h. In a choice setting, consistent preference of Diet A over Diet D was observed within 7 h followed by stabilisation at about 65% diet A to 35% diet D. 5. Chicks fed diet D alone from 8 to 20 d of age, then placed in the same choice situation preferred diet A to D with a delay of less than one h and stabilisation at about 85%. Chicks provided a choice of diets A and D from 8 to 20 d, and then diet D alone reduced their food intake more quickly than those not given a choice initially. 6. Broiler chicks appear to react to amino acid deficiencies within a short period (hours) by adjusting their feed intake and/or selection. The response is influenced by age and prior experience. PMID- 8242410 TI - Lysine requirement of broiler chicks as affected by protein source and method of statistical evaluation. AB - 1. An experiment was designed to test if the lysine requirement, expressed as g lysine/kg CP, was the same for several protein sources. 2. Groundnut meal, groundnut meal adjusted with indispensable amino acids or sesame meal supplied the dietary CP at 180 g/kg diet. Increments of lysine (1.5 g/kg diet) were added to each of these diets. 3. The gain, food intake and food efficiency responses of broiler chicks were analysed using a quadratic equation and a two-slope method. An estimate of lysine requirements was also obtained from a survey of college students. 4. The different methods produced widely different estimates of lysine requirement. 5. The average lysine requirement was estimated at 50.1 g lysine/kg CP for groundnut meal, 61.7 for adjusted groundnut meal and 54.9 for sesame meal. 6. Reasons for the effect of statistical analysis and protein source on lysine requirement are discussed. PMID- 8242411 TI - Effects of genotype and cryopreservation of avian semen on fertility and number of perivitelline spermatozoa. AB - 1. The fertility of freshly diluted and cryopreserved samples of semen obtained from a population of chickens selected for duration of fertility of cryopreserved spermatozoa (FS line) and its unselected control (FC line) were compared over a range of spermatozoa concentrations (10, 40, 80, and 160 x 10(6) sperm/50 microliters insemination). 2. The spermatozoa of the FS line had greater fertility than spermatozoa of the FC line, whether freshly diluted or cryopreserved. Cryopreservation resulted in a reduction in fertility, regardless of line. There were no significant line by genotype interactions. 3. There were fewer spermatozoa from the FC line than the FS line found in the perivitelline membrane (perivitelline spermatozoa). The increase in number of perivitelline spermatozoa with increasing sperm concentration was greater in the FS than FC line. However, the slope of the increase in sperm number in the perivitelline membrane with increasing concentrations of cryopreserved spermatozoa was zero. 4. A minimum of 10(3) perivitelline spermatozoa must be found on day 2 post insemination for duration of fertility to exceed three days. The ability to produce spermatozoa capable of reaching the forming perivitelline membrane appears to be a quantitative, rather than a qualitative, trait and may be subject to genetic manipulation. PMID- 8242412 TI - Induced moulting in cockerels: effects on sperm production, plasma concentrations of luteinising hormone, testosterone and thyroxine, and on pituitary sensitivity to luteinising hormone-releasing hormone. AB - 1. The ability of a moult-inducing procedure to restore high levels of sperm production was assessed, in two experiments, using cockerels with reduced sperm production. The moulting procedure consisted of a period of food and light restriction for 6 weeks. The moulted birds were compared with control birds for 20 weeks. 2. Moult induction resulted in decreased daily sperm output (DSO) and plasma testosterone concentration, from weeks 3 to 7. In the first experiment, plasma luteinising hormone (LH) concentration in moulted birds was reduced on week 5. 3. No change in pituitary sensitivity to chicken luteinising hormone releasing hormone-I (cLHRH-I) was detected at week 3 in moulted cockerels. In moulted birds, a transient increase in plasma thyroxine concentration was detected. 4. After the end of moult induction, testosterone concentrations increased, plasma LH showed a rebound at week 7 and the pituitary sensitivity to LHRH was increased at week 9. 5. This increased activity of the pituitary testicular axis resulted for a short time in an increase in DSO of moulted birds compared with that of controls. Although amelioration was moderate, this result indicates the possibility of improving sperm production in the cockerel by using an induced moulting procedure. PMID- 8242413 TI - Modification of intra-ovarian follicular distributions in broiler breeder hens by ad libitum or restricted feeding. AB - 1. Ovarian follicles in broiler hens from 3 lines fed ad libitum or food restricted were examined at point of lay and from 2 of these lines at 47 weeks of age. The lines were Ross 1 birds and birds selected to be lean or fat on the basis of their plasma very low density lipoprotein concentrations at 7 weeks of age. 2. Follicles of diameter greater than 1.4 mm were classified into size groups, and the numbers in these groups in the anterior and posterior halves of the ovary were compared for the effects of line and feeding regimen. 3. There were more yellow follicles (> 8 mm diameter) in the anterior half of the ovary, and this difference was greater for ad libitum birds. Ad libitum feeding did not influence the number of yellow follicles in the posterior segment of the ovary. 4. The increase in numbers of yellow follicles in the anterior segment of the ovary differed according to line at point of lay, but not at 47 weeks of age. 5. Birds of 47 weeks of age showed higher numbers of follicles 1.4-6 mm in diameter in the posterior segment of the ovary, and smaller numbers of follicles 6-8 mm in diameter. Thus, although large numbers of white follicles were in the earlier stages of growth, there were fewer ovulable yellow follicles because of higher levels of atresia. PMID- 8242414 TI - Blood flow distribution during graded treadmill exercise in domestic cockerels. AB - 1. White Leghorn cockerels were exercised continuously for 45 minutes on a treadmill at work loads equivalent to 2.1 and 2.7 times pre-exercise metabolic rates, the latter being close to the performance limits of the experimental birds. 2. Relative blood flow to the abdominal viscera and selected leg muscles was estimated using 113Sn-, 153Gd- and 46Sc-labelled microspheres, injected into the root of the aorta. 3. There was no significant shift of blood flow away from the viscera towards the leg muscles at either work load. However, relative colonic and duodenal blood flow fell significantly at both work loads and relative pancreatic blood flow doubled at the higher work load. 4. It is concluded that, as previously reported in female domestic fowl performing physical exercise close to their performance limits, male fowl are largely refractory to shifts in blood flow away from the viscera to the working muscles. Nevertheless, there is evidence of blood flow shifts between visceral organs, probably related to the intermediary metabolism of fuel substrates. PMID- 8242415 TI - Effects of body weight at sexual maturity and the degree and age of restriction during rearing on the ovarian follicular hierarchy of broiler breeder females. AB - 1. Groups of 18 birds were reared to 10 different body weight targets in a randomised block experiment to assess the effect of body weight and the degree and age of restriction on ovarian function at first egg. 2. There was no effect of age at restriction (15 to 18 compared with 19 to 22 weeks) on the number of yellow follicles. 3. The treatment with the larger body weight of two groups given the same degree of restriction (maintenance) had more follicles than the treatment which had a smaller body weight. 4. There was a positive linear relationship between the number of yellow follicles and body weight and food intake. Addition of the weight of fat or age at first egg to the model increased the residual mean squares. 5. The proportion of atresia among yellow follicles and the incidence of internal ovulation increased with body weight at first egg. 6. It was concluded that the physiological processes occurring during rearing which underlie ovarian function are reflected solely in the body weight and food intake of broiler breeders at first egg. PMID- 8242416 TI - Effect of thyroxine on testicular function, circulating luteinising hormone and pituitary sensitivity to luteinising hormone-releasing hormone in the cockerel (Gallus domesticus). AB - 1. The effect of thyroxine (T4) on reproductive function in the adult cockerel was followed for 11 weeks. Broiler cockerels aged 96 weeks were fed on diets containing either 0, 2 or 5 mg T4/kg for 4 weeks. 2. Daily sperm output (DSO) was significantly reduced (P < 0.01) in the T4-treated groups compared with that of controls at weeks 5 and 7. In the group given 5 mg T4/kg, plasma testosterone concentration was significantly reduced (P < 0.01) compared with that in controls during the T4 treatment, in spite of the fact that there was a decrease in concentration in both control and experimental birds. Plasma luteinising hormone (LH) concentration was significantly decreased (P < 0.05) in both of the groups given T4 treatments after 3 weeks. 3. Plasma testosterone concentrations and DSO had returned to control values at weeks 5 and 11 respectively, while plasma LH showed a transient but significant (P < 0.001) rebound after removal of thyroxine from the food. 4. In contrast to other variables, the pituitary responsivity to cLHRH-I injections, was not decreased during the feeding of the T4 diet but was, on the contrary, significantly increased (P < 0.05) during treatment with 5 mg T4/kg diet, and after the end of the treatment with 2 mg T4/kg diet. 5. These results provide some evidence for an inhibitory effect of large doses of T4 on the reproductive function in the adult cockerel. Although the possibility of a direct effect of T4 on the testes cannot be excluded, T4 is likely to act, at least in part, at the hypothalamo-pituitary level, and not through a reduction in the pituitary sensitivity to LHRH. PMID- 8242417 TI - Alpha-tocopherol absorption and polyunsaturated fatty acid metabolism in the developing chick embryo. AB - 1. A study has been made relating alpha-tocopherol uptake from the yolk and yolk sac membrane to the polyunsaturated fatty acid status of the liver during incubation. 2. Whereas at day 15 of incubation lipid in the liver showed an exceedingly high degree of unsaturation and potential for oxidation because of accumulation of C20 and C22 polyunsaturated fatty acids, alpha-tocopherol concentrations were exceedingly low. 3. In contrast, very high concentrations of alpha-tocopherol occurred within the liver just before hatching by which time amounts of polyunsaturates and the need for oxidation protection had reduced considerably. PMID- 8242418 TI - Comparison of calpain and calpastatin activities in skeletal muscle of broiler and layer chickens. AB - 1. The objective of this study was to estimate the difference between broiler and layer chicks in the activities of calpain and calpastatin (inhibitor of calpain) in breast muscle. Differences between broilers and layers in body weight, daily gain at 3 weeks of age and fractional growth rate (FGR) during 2 and 3 weeks of age were statistically significant (P < 0.01). 2. Calpain and calpastatin activities were measured at three weeks of age with alkali-denatured casein as a substrate. The m-calpain (calpain activated by millimolar calcium concentration) activities in units/g muscle and units/mg extractable muscle protein were 0.779 and 0.353 for broilers, and 1.042 and 0.440 for layers, respectively. The calpastatin activities in units/g muscle and units/mg extractable muscle protein were 0.332 and 0.153 for broilers, and 0.262 and 0.112 for layers, respectively. 3. Broilers with high FGR showed low m-calpain and high calpastatin activities. In contrast, layers with low FGR showed high m-calpain and low calpastatin activities. 4. These results suggest that m-calpain and calpastatin activities in skeletal muscle vary between breeds which have different rates of muscle production. PMID- 8242419 TI - [Membrane receptors and coupling proteins in adenohypophyseal cells]. AB - Dopamine receptors of D2 type present on lactotroph cells are coupled to a large series of transduction mechanisms. Beside their negative coupling with adenylate cyclase, they are also coupled with potassium and calcium channels, leading to a decreased intracellular calcium concentration. In addition, D2 dopamine receptors also modulate phospholipase activities. Dopamine inhibits inositol phosphate production, through two distinct mechanisms. One of them could represent a direct negative coupling with phospholipase C. All these transduction mechanisms of the D2 dopamine receptors implicate G proteins sensitive to pertussis toxin. In contrast, these receptors are negatively coupled to phospholipase A2 through G proteins insensitive to this toxin. Both isoforms of the D2 dopamine receptor, generated by alternate splicing of a single gene are present in lactotroph cells. After transfection in CH4C1 cells the two isoforms are coupled with adenylate cyclase while only the shortest isoform appears negatively coupled to phospholipase C. Functional D2 dopamine receptors are present in human prolactinomas. Resistance to bromocriptine therapy is associated with a decrease density of these receptors in the tumor. In addition, the ratio of the two receptor isoforms (measured by PCR) is different in responsive and resistant tumors. Furthermore, the activity of Gi/Go proteins coupled to adenylate cyclase appears also affected in resistant tumors. Resistance to bromocriptine therapy appears thus to involve multiple changes at the different levels of the multiple mechanisms of action of dopamine on lactotroph cells. PMID- 8242420 TI - [Elements concerning the cellular organization of glycine receptors dependent of chlorine]. AB - Glycine is one of the main inhibitory neurotransmitter in the central nervous system of vertebrates where it acts by activating a chloride conductance. The distribution of glycine receptor at the neuronal surface was analysed by immunocytochemistry with monoclonal antibodies raised against the purified receptor. In the rat spinal cord as well as in other areas of the central nervous system, these receptors are localized at the postsynaptic membrane and are concentrated in front of the presynaptic release sites. Thus, they define functional microdomains at the plasma membrane. A similar organisation was observed in a motor command neuron: the Mauthner cell of Teleosts. Further, in this model, a quantitative analysis using confocal microscopy has established that the postsynaptic microdomains are arranged according to a somatodendritic gradient with the larger clusters at the tip of the dendrites. The use of primary cultures of rat or mouse spinal cord neurons has allowed to study the ontogenesis of the glycine receptor. We have shown that these receptors are, here also, organized in clusters present at the neuronal surface and that the formation of these aggregates occurs simultaneously to the establishment of synaptic contacts. PMID- 8242421 TI - [Mechanism of cell death in neurodegenerative disorders: apropos of Parkinson disease]. AB - Parkinson's disease is a neurodegenerative disorder characterized by a selective loss of nerve cells which die slowly, but more rapidly than similar effects due to aging. This premature nerve cell death results from genetic and environmental factors, normal aging being probably an accessory mechanism. The cause of dopaminergic nerve cell death in Parkinson's disease includes an oxidative stress resulting from an overproduction of free radicals, a decreased metabolism of the mitochondrial respiratory chain, a decreased defence against calcium, and many other putative mechanisms. The fact that there are still several hypothesises of cell death in Parkinson's disease indicates that we are still far from discovering the cause of the disease. PMID- 8242422 TI - [Growth and survival factors of spinal motoneurons]. AB - The development of motoneurons in the spinal cord is strongly dependent on their interactions with their target tissue, skeletal muscle, and with other cells of the central nervous system. The molecular nature of these interactions has remained obscure for many years. However, over the last few years, known growth factors have been shown to have biological activity on the survival of motoneurons, at least in culture. The factors that have been studied are members of the FGF family (fibroblast growth factors), the TGF-beta family (transforming growth factor-beta), CNTF (ciliary neurotrophic factor) and CDF-LIF (cholinergic development factor-leukaemia inhibitory factor). There are also strong reasons to suppose that at least one member of the neurotrophin family (the family that contains Nerve Growth Factor) is involved in motoneuron development. A more detailed analysis of the biological role of each of these factors should not only enlighten us as to the importance of cell-cell interactions in development of the motoneuron, but also open the way to attempts to slow motoneuron death in pathological situations, either in animals or in man. PMID- 8242423 TI - [Functional properties of tachykinin receptors in the central nervous system]. AB - Three types of tachykinin receptors (NK1, NK2 and NK3) have been identified, their endogenous ligands being substance P, neurokinin A and neurokinin B respectively. Peptide agonists and non peptide antagonists are now available allowing to study the functional properties of these receptors which are coupled to G proteins. Depending on the tissue, stimulations of phospholipase C occur with NK1, NK2 or NK3 agonists. Peptide, a C-terminal substance P analog and substance P(1-7) seem to act on tachykinin receptors distinct from those of the NK1, NK2 or NK3 type. NK2 binding sites have not been visualized in the brain. Nevertheless, biological responses of the NK2 type have been shown both in the substantia nigra and the striatum in studies on dopamine release. PMID- 8242424 TI - [Transcriptional regulation of an enzyme involved in the synthesis of melatonin]. AB - The indolic hormone melatonin is produced by the pineal gland according to a daily rhythm. The terminal step of melatonin synthesis is catalyzed by hydroxyindole-O-methyltransferase (HIOMT, EC 2.1.1.4). Adaptation to constant light or darkness modifies HIOMT activity and concentration. Using a cDNA probe encoding HIOMT, we investigated the effect of environmental lighting on HIOMT gene expression in the chicken pineal gland. HIOMT mRNA levels increased by 100% in constant light as compared with constant darkness. In addition, the present study disclosed the existence of a day/night rhythm of HIOMT gene transcription, with 3-fold higher mRNA levels at midday than at midnight. Unexpected darkness did not prevent the daytime rise in HIOMT mRNA levels, whereas unexpected light prevented the night-time fall in HIOMT mRNA levels. Together, the data would suggest that the day/night rhythm of HIOMT gene transcription in the chicken pineal gland involves both a response to light and the activity of a biological oscillator. PMID- 8242425 TI - [Photoreceptor cells of the pineal body in culture: effect of 17 beta-estradiol on the production of melatonin]. AB - Pineal photoreceptor cells produce the neurohormone, melatonin, a major "Zeitgeber" of the organism. This compound has been involved in the control of development, growth, sexual maturation, and seasonal reproductive cycles. We investigated, here, the effects of estradiol-17 beta on melatonin production by cultured pineal photoreceptor cells. Under a light/dark (LD: 12/12) cycle, cultured trout pineal cells maintained a rhythmic secretion of melatonin with higher amounts being released during the dark phase. The amplitude of the rhythm tended to increase with time spent in culture. Application of estradiol-17 beta during the dark phase of a LD cycle (i.e., for 12 h) affected melatonin release in a dose-dependent manner: low concentrations (10(-10) to 10(-8) mol/l) were inhibitory whereas high concentrations (over 10(-7) mol/l) were stimulatory. When estradiol-17 beta was applied continuously for several 24 h LD cycles, the inhibitory effect observed during the first dark phase disappeared later on. Rather, in the presence of estradiol-17 beta, at the concentrations of 10(-9) and 10(-6) mol/l, a high amplitude rhythm in melatonin secretion was recovered faster, when compared to controls. Replacement of 10% fetal calf serum by 0.1% bovine serum albumin did not affect the responses of the photoreceptor cells to 10(-6) mol/l of estradiol-17 beta. The present results bring the first evidence that estradiol-17 beta modulates melatonin secretion by cultured pineal photoreceptors. They further support the idea according to which sexual hormones exert a feedback regulation on the pineal. Pineal photoreceptors appear as multieffector cells which transduce information from, both, physical (photoperiod) and internal (chemical) factors. PMID- 8242426 TI - [Antigenic changes of Tau protein induced by glutamate on primary cultures of neurons: immunocytochemistry study]. AB - Degenerating neurons in Alzheimer's disease are characterized by the presence of neurofibrillary tangles constituted by paired helical filaments (PHF). Abnormally phosphorylated Tau protein, a microtubule associated protein is one of the major component of PHF. Abnormal phosphorylation seems to be located in the C-terminal domain but also in the N-terminal region of Tau proteins. Previous studies demonstrated that calcium-mediated glutamate toxicity produces a dose-dependent increase of Tau immunolabellings in neuronal cultures. Biochemical results revealed that these changes could be associated with abnormal Tau migrations on immunoblots. Using three anti-Tau antibodies the present study shows that glutamate toxicity induces in neuronal cultures, Tau modifications localized in the N- and C-terminal domains of the protein. These findings suggest the possibility that glutamate toxicity can induce Tau antigenic changes involving probably the whole molecule. PMID- 8242427 TI - In vitro cytotoxicity test for estimating non-ocular irritation dose of ophthalmic solutions. AB - The in vitro cytotoxicity test for estimating the non-ocular irritation dose of ophthalmic solutions was investigated. In the in vitro test, normal human epidermal keratinocytes (NHEK) in a confluent monolayer were incubated for 48 hr in a medium with test compounds. The concentration of a test compound which causes a 50% reduction in NHEK viability was determined as IC50 by MTT colorimetric assay. For comparison, the in vivo rabbit ocular irritation tests were carried out by the standard Draize method. The maximum concentration, which did not show any ocular irritation, was determined as DS0. The results showed the correlation coefficient between the IC50 values and the DS0 values for 19 test compounds to be 0.82. However, the correlation coefficients for 10 compounds, which have IC50 values of less than 300 micrograms/ml, and for 7 alcohols were 0.99. The IC50-DS0 correlation curves obtained could be utilized as the critical concentrations for ocular irritation. These results suggest that our in vitro/in vivo test can estimate non-ocular irritation dose of the ophthalmic preparations in advance of the in vivo tests. PMID- 8242428 TI - Two different pathways for necrotic cell death induced by free radicals. AB - Plasma membrane modifications have been widely recognized as crucial factors in cell injury and death. One of these modifications, surface blebbing, has been considered as an injury-marker associated with a series of biochemical and physiological modifications. Our study focused on the different effects of free radical-induced cell damage by quinone menadione (2-methyl-1,4-naphthoquinone) and by hyperthermic shock (45 degrees C) on the erythroleukemic cell line K562. Different techniques including immunofluorescence, freeze-fracturing, and electron paramagnetic resonance spectroscopy were employed. Menadione induced the formation of surface blebs, accompanied by a rearrangement of the microfilament system and changes in the distribution of plasma membrane proteins. In contrast, heat-shocked cells showed neither blebbing nor important cytoskeletal changes. Finally, the electron paramagnetic resonance results showed an increase in membrane order not specifically related to the type of free radical-induced stress. These cell death features appear to suggest the existence of two different types of pathways for necrotic cell death: both treatments induce cell injury and eventual death by modifying plasma membrane integrity and function. However, one involves cytoskeleton-dependent surface blebbing, whereas the other does not. PMID- 8242429 TI - The carcinogen benzo(e)pyrene is metabolized by DM15 cells without an uncoupling effect on their gap junctions. AB - Benzo(e)pyrene (B(e)P) promotes carcinogenesis in the skin. Unlike some other promoters however, B(e)P does not produce an uncoupling effect on gap junction permeability in DM15 transformed fibroblasts. This study demonstrates that DM15 cells exhibit a relatively high level of B(e)P metabolism. Moreover, although pretreatment of DM15 cells with benz(a)anthracene results in an 8-fold increase of arylhydrocarbon hydroxylase activity and a 2-fold increase in the rate of B(e)P metabolism, it did not enable B(e)P to affect Lucifer Yellow transfer between DM15 cells. We conclude that neither B(e)P nor its metabolites are capable of uncoupling gap junction permeability in DM15 cells. PMID- 8242430 TI - Genistein inhibits calcium release by platelet-derived growth factor but not bradykinin or cadmium in human fibroblasts. AB - Cd2+ provokes inositol trisphosphate production and releases stored Ca2+, apparently by binding to a zinc site in the external domain of an orphan receptor. One microM Cd2+ evokes an immediate spike in cytosolic free Ca2+, which is similar to that evoked by bradykinin. Platelet-derived growth factor (PDGF) also increases free Ca2+ in human dermal fibroblasts, but there is a distinct lag before free Ca2+ rises in response to PDGF. Genistein, which selectively inhibits tyrosine kinases, markedly inhibited Ca2+ mobilization evoked by PDGF. Calcium mobilization triggered by cadmium or bradykinin was relatively insensitive to genistein. The PDGF receptor is known to be a tyrosine kinase, which phosphorylates and thereby activates phospholipase C gamma, whereas a G protein couples the bradykinin receptor to another phospholipase C isoform. These findings support the hypothesis that the orphan receptor triggered by cadmium is coupled to phospholipase C via a G protein. PMID- 8242431 TI - Fibronectin turnover in human mesangial cell cultures as affected by adriamycin. AB - Fibronectin (FN) turnover and turnover changes induced by the anticancer drug Adriamycin (ADR) were measured in human mesangial cells (HMC) in vitro. HMC cultures synthesize cellular FN (2.2 +/- 0.3% of total protein synthesis; n = 12) which is secreted and incorporated into a fibrillar extracellular matrix (ECM). A 24 hr incubation of HMC with ADR (0.5-5 micrograms/ml) resulted in an accumulation of FN in the culture medium, with a maximum increase following 5 micrograms/ml (7.3 +/- 2.3 pg/cell vs. controls: 4.4 +/- 1.9 pg/cell; n = 10). Correspondingly, radioactively labeled immunoprecipitable FN was increased in a dosage-dependent manner in the culture medium up to 50% vs. controls. The incorporation of radioactively labeled FN into ECM was significantly increased following 2 micrograms ADR/ml. In accordance, immunofluorescence staining revealed an expansion of pericellular FN fibers in cultures exposed to 2 micrograms ADR/ml. Concomitant with the accumulation of extracellular FN, radioactively labeled FN in the cells was reduced by 22%. Qualitative characterization of FN patterns revealed a diminished number of degradation products in the culture medium of ADR-treated HMC. These data suggest that ADR interferes with the turnover of FN secreted by HMC in vitro in such a way that FN accumulates extracellularly. This in turn leads to a reduced FN synthesis. These findings are compatible with a loss of urinary FN degradation products accompanying the onset of proteinuria in ADR-treated rats. PMID- 8242432 TI - Effects of specific inhibitors of cellular functions on sulfur mustard-induced cell death. AB - This study was conducted to determine whether inhibitors of normal cellular functions can reduce cytotoxicity induced by sulfur mustard (HD). The compounds examined include inhibitors of poly(ADP-ribose) polymerase (PADPRP), inhibitors of mono(ADP-ribose) transferase (MADPRT), inhibitors of lipid peroxidation, and an inhibitor of protein synthesis. To determine the effects of these compounds on HD-induced cell death, human lymphocyte preparations were treated with known concentrations (0.1 microM to 1000 microM) of an inhibitor and exposed to an estimated 87% effect concentration (EC87) of HD (170 microM) for loss in cell viability. Cell viability was determined at 24-26 hr post-exposure to HD using a dye (propidium iodide) exclusion assay and a flow cytometer. All of the selected PADPRP inhibitors were found to be effective at reducing the cytotoxic effects of HD. These inhibitors were rank-ordered based on the concentration that gives 50% (EC50) reduction of HD-induced cell death. A significant correlation (r = 0.94) was observed between the compounds' ability to inhibit PADPRP and the compounds' ability to reduce HD- induced cell death, suggesting that PADPRP plays a role in HD-induced cell death. Inhibitors of MADPRT, lipid peroxidation, and protein synthesis were not effective at reducing HD-induced cell death. PMID- 8242433 TI - Induction of the human growth hormone gene placed under human hsp70 promoter control in mouse cells: a quantitative indicator of metal toxicity. AB - An in vitro test method for general metal toxicity screening was designed, based on the cellular response to stress. The expression of a transfected human growth hormone gene sequence driven by the human heat-shock protein 70 promoter in NIH/3T3 cells was used as marker of noxious contact with metal compounds. Out of a series of 31 metals, 17 were competent for inducing this stress response system. According to the effective concentration and to the intensity of the response, three different clusters of positive compounds emerged and were ranked as strong, intermediate strength and weak inducers. These results correlated well with data from other in vivo and in vitro metal toxicity studies, including LD50 in mice. Apparently the positive/negative compounds also fitted well with data from genotoxicity and carcinogenesis studies on metal salts. PMID- 8242434 TI - Effects of fenvalerate and esfenvalerate on hepatic gap junctional intercellular communication in rats. AB - Effects of in vivo exposure with fenvalerate, esfenvalerate and DDT on hepatic gap junctional intercellular communication (GJIC) in Sprague-Dawley (SD) rats were examined by in vivo/in vitro dye-transfer assay and by immunohistochemical staining of connexin 32 (Cx32, major liver gap junction protein). Fenvalerate (75 mg/kg/day), esfenvalerate (25 mg/kg/day), DDT (50 mg/kg/day) and corn oil (vehicle control, 5 ml/kg/day) were administered orally once a day. Animals were killed at weeks 1, 2, 4 and 6 after starting the experiment. In the fenvalerate- and esfenvalerate-groups, no compound-related changes in GJIC and Cx32 expression were observed. On the contrary, in the DDT-group, average sizes of the dye spread after injection of Lucifer Yellow decreased at weeks 1, 2 and 4, and the area per GJ spot shown by Cx32-immunohistochemical staining decreased at weeks 4 and 6. It is concluded that neither fenvalerate nor esfenvalerate inhibits hepatic GJIC with in vivo exposure. PMID- 8242435 TI - Hodgkin's disease. PMID- 8242436 TI - Hodgkin's disease. AB - The etiology of Hodgkin's disease is unknown, and the identity of the normal counterpart of the malignant cell involved in Hodgkin's disease remains controversial. Yet, three of every four patients diagnosed with Hodgkin's disease will be cured. This success in treatment is due in large part to the application of modern oncologic techniques, such as combination chemotherapy and the use of staging to guide therapy. This article reviews important advances in the understanding and treatment of Hodgkin's disease. PMID- 8242437 TI - Natural history and clinical behavior of in situ carcinoma of the human urinary bladder. 1964. PMID- 8242438 TI - Report on the UICC workshops on breast cancer screening in premenopausal women in developed countries. Union Internationale Contre le Cancer. AB - At the recent UICC Meeting on Breast Cancer Screening in Premenopausal Women, three workshops were held to address issues concerning the evaluation of the efficacy of screening, current strategies in research and practice, and future directions in breast cancer control. The authors report on the results of these workshops, including a proposal for a multinational clinical study of breast cancer screening in premenopausal women. PMID- 8242439 TI - On humanizing the critical care environment. AB - Critical care unit design should strive to provide physical and psychologic comfort for patients, families, and staff members. Thoughtful and creative analysis of the critical care unit milieu will result in both functional and environmental improvements. Clinicians, architects, engineers, interior designers, and equipment manufacturers should collectively focus their efforts on the integration of technology and comfort in this stressful setting. PMID- 8242440 TI - Critical care unit design: a collaborative approach. AB - Determining a critical care unit design can be a complex and sometimes overwhelming task. Even so, through group participation the project can be an opportunity for developing collaborative relationships and having fun as well. Collaborative decision making can prevent unnecessary duplication, promote greater staff satisfaction, and improve overall cost containment. The article describes the collaborative approach used at Elizabeth Community Health Center in Lincoln, Nebraska to plan the design of the critical care unit, staff orientation, the open house, and the move process. PMID- 8242441 TI - Design considerations for heating, air conditioning, and exhaust systems in critical care units. AB - Air quality, temperature control, and air movement directly affect infection control, comfort, and safety in critical care units. This article explains basic concepts in the heating, ventilation, and air conditioning that affect patient care. Functional requirements must be determined early in the design process and discussed with clinicians, engineers, and architects to assure understanding and correct design. PMID- 8242442 TI - Impact of technology of the critically ill: the utilization of mechanical ventricular assist devices. AB - The advancement of high technology in critical care has meant refined techniques in cardiac surgery, new pharmacologic agents, and promises for longer, higher quality lives for many cardiac patients. This article describes the use of one of the most important technologic devices to assist the failing ventricle, the ventricular assist device. Topics addressed include historical perspective of the ventricular assist device, its indications and contraindications, the types of devices, potential complications, nursing implications, and a case study of a male patient with a ventricular assist device. PMID- 8242443 TI - Toward a hierarchy of adaptation to biomedical technology. AB - The article attempts to derive a hierarchy of patient adaptation for a variety of technologic devices that are used in medical and nursing care. A review of related literature on some existing taxonomies of biomedical technology and an empiric foundation based on Roy's Adaptation Model suggest that patient adaptation to biomedical devices is affected by at least two factors--the degree of invasiveness and automation--that are inherent in all bedside devices. It appears that patients' need for adaptation to technology is more apparent as automation decreases and invasiveness increases. Further research is necessary to determine whether this relationship holds for all types of technology and to identify additional factors that might contribute to the development of an adaptation hierarchy for technologic devices. The nurse's role as a mediator of stress between the patient and technology is also explored. PMID- 8242444 TI - Concepts in headwall selection and design. AB - The headwall is the interface between the patient and critical care technology. This article lists headwall components, outlines various types of headwall systems, and discusses factors in selecting the type of headwall to support the desired model of care. PMID- 8242445 TI - Perceived needs of the family members in the critical care waiting room. AB - Family needs during the critical care experience of an adult member was the focus of this descriptive exploratory study conducted with 94 family members. The study describes the perceived needs of family members during the first 18 to 24 hours after admission of a patient to the critical care unit. The article further identifies the order in which the family perceived those needs to be met 36 to 48 hours after admission of that patient. Family members of adult patients completed a three-part instrument, which consisted of the Demographics Data Questionnaire, the Critical Care Family Needs Inventory (CCFNI), and the Needs Met Inventory (NMI). Items with which the family strongly agreed are discussed. The correlation between the CCFNI and the NMI is explicated. Also, the unusual finding of an inverse relationship between education and comfort/support statements is presented. PMID- 8242446 TI - Experiences in children of critically ill parents: a time of emotional disruption and need for support. AB - The purpose of this phenomenologic study was to describe effects of a parental critical care hospitalization on the family as viewed by children. Interviews were conducted with eleven male phenomenologic techniques and showed these four major themes: (1) emotional turmoil, (2) family disruption, (3) need for support, and (4) minor illnesses. These disruptions are strong and need to be treated through nursing interventions. PMID- 8242447 TI - If they don't ask these questions, give them the answers. AB - Clinicians must be active rather than passive participants in the design process. This article lists basic concepts that pertain to the model of care and are the foundation of the design. Clinicians must be in agreement on these concepts and discuss the rationale for them with the design team to establish the project goals. PMID- 8242448 TI - Using mixed venous oxygen saturation data to trend overall oxygenation in cardiothoracic surgical patients. AB - Cardiothoracio surgery patients routinely have pulmonary artery catheters with mixed venous oxygen saturation fiberoptios placed in the operating room. This monitoring capability allows for a continual assessment of the oxygen supply demand balance. Critical care nurses are encouraged to use mixed venous oxygen saturation trends to the fullest potential. This is possible through the integration of oxygenation and hemodynamic data in both patient assessments and monitoring therapies. PMID- 8242449 TI - Economics of mastitis and mastitis control. AB - Mastitis is the most costly disease of dairy cows. The major economic loss of all forms of mastitis results from reduced milk production. Because of the difficulty in controlling environmental mastitis organisms, mastitis will maintain this role in the foreseeable future. PMID- 8242450 TI - Epidemiologic methods in mastitis treatment and control. AB - Methods and concepts of epidemiology offer means whereby udder health can be monitored and evaluated. Prerequisite to a sound epidemiologic approach is development of measures of mastitis that minimize biases and that account for sensitivity and specificity of diagnostic tests. Mastitis surveillance offers an ongoing and passive system for evaluation of udder health, whereas clinical and observational trials offer a more proactive and developmental approach to improving udder health. PMID- 8242451 TI - Procedures for mastitis diagnosis and control. AB - Procedures for mastitis diagnosis and control include culturing individual cow and bulk tank milk samples, antibiotic susceptibility testing, and evaluation of somatic cell count reports and clinical mastitis treatment records. Integrated use of such procedures is necessary for effective mastitis diagnosis and control. PMID- 8242452 TI - Microbiologic examination of bulk tank milk. AB - Microbiologic examination of bulk tank milk can be a useful technique in assessment of mastitis and milk quality. Sample collection and handling, laboratory techniques, and interpretation and use of bulk tank microbiology are discussed. Recommendations are made based upon a survey of mastitis laboratories and the author's experience. PMID- 8242453 TI - Contagious mastitis. AB - Contagious mastitis is defined. The major mastitis pathogens are Streptococcus agalactiae, Staphylococcus aureus, Corynebacterium bovis, Mycoplasma sp, and Streptococcus dysgalactiae. These pathogens are discussed relative to prevalence, virulence factors, pathology, and control. These control measures include milking time hygiene, segregation, culling, vaccination, and treatment. PMID- 8242454 TI - Environmental mastitis. AB - Environmental mastitis affects all dairy farms and generally is the major mastitis problem on modern, well managed dairy farms. Control measures effective against contagious pathogens are of little value in controlling of environmental pathogens. Control of environmental mastitis is achieved by reducing exposure of teat ends to environmental pathogens and by maximizing the resistance of the cow to intramammary infection. Significant sources of environmental pathogens are organic bedding materials, manure covered alleyways, and wet or damp areas in barns, exercise lots, or pastures. Milking time hygiene can influence teat-end exposure. In general, exposure is minimized when all areas of the environment are clean, cool, and dry. Resistance is maximized by providing a stress-free environment that minimizes teat-end injury, and by feeding balanced diets sufficient in vitamin E and selenium. Antibiotic therapy during lactation or the dry period is of little value in the control of environmental mastitis in dairy herds, with the exception of preventing environmental streptococcal infection during the early dry period. Effective vaccines may help reduce the impact of environmental mastitis in the near future. PMID- 8242455 TI - Advances in the therapy for mastitis. AB - Methods to enhance mammary resistance to bacterial infection and to reduce the effects of existing infections without the use of antimicrobial agents are becoming more attractive, primarily because of increasing pressure from consumers and regulatory agencies to decrease the risk of drug residues in milk. Because of the difficulty in obtaining satisfactory results with existing drug formulations, new approaches in the treatment of mastitis should emphasize better understanding of mammary gland pharmacokinetics, ameliorating the pathologic effects of infection, and enhancing natural defenses. Efficacy studies should emphasize milk production and long-term survival of cows to allow economic evaluations. PMID- 8242456 TI - Evolution of udder hygiene. Premilking teat sanitation. AB - Compared with post-milking teat dipping, predipping is in its infancy. Few controlled studies have been published on premilking teat disinfection/sanitation. Experimental challenge procedures resulted in consistent reductions for predipping compared with no udder preparation and conventional udder preparation, indicating a potential for effectiveness. Challenge studies that included the treatments of predip only and postdip only indicated that interactions during the milking process require elucidation. Positive benefits have been observed for predipping in field trials among some herds, but wide variation has been observed between herds in all field trial evaluations. Parity, DIM, and season of the year had major interactions on efficacy of predipping in Pankey et al's 1987 report; other trials have been of shorter duration and precluded analysis of these interactions. Interactions among these variables influenced incidence of mastitis by environmental pathogens. The length of time after milking before teats are contaminated probably is a major influence on predip efficacy. Predipping has reduced incidence of new IMIs and new cases of clinical mastitis. Unfortunately, certain factors negated these positive effects. These factors need to be defined. Producers should monitor effects of predipping to determine whether the investment in product and time has an economic return under the conditions of their dairy. PMID- 8242457 TI - Milking machine function and analysis. AB - Milking system evaluation is facilitated by use of a pulsator recorder, airflow meter, and a vacuum stability gauge. The most important points of a milking system are vacuum, inflations, and pulsation. An evaluation sheet for milking systems as proposed by the National Mastitis Council is presented. Paramount to evaluation of any milking system is that the parameters must relate to milking performance or teat and udder health. PMID- 8242458 TI - Immunization and immunotherapy for mastitis. AB - Immunization and immunotherapy for mastitis are active areas of investigation. The past decade has seen development of effective and economical R-mutant vaccines for gram-negative mastitis. These vaccines doubtless will prove beneficial on well managed dairies that have eradicated contagious mastitis pathogens. Development of vaccines for other mastitis pathogens has been noticeably slower. A commercially available Staphylococcus aureus vaccine appears to reduce the frequency and severity of clinical episodes, but probably has minimal impact on the incidence or prevalence of infection. This product has not been extensively studied. The recent recognition of virulence factors produced in vivo by Staphylococcus aureus may provide a breakthrough in the development and production of Staphylococcus aureus vaccines. Bacterins employing this principle presently are not commercially available, however. In the case of all contagious mastitis pathogens (Streptococcus agalactiae, Staphylococcus aureus, and Mycoplasma spp.), traditional control and eradication efforts (teat dip, dry cow therapy, culling programs) likely will prove preferable to long-term immunization. Ongoing research may provide more efficacious vaccines for these mastitis syndromes. Immunostimulants are an active area of research. Although leukopoietic factors appear promising as immunostimulants, no compound has clearly demonstrated efficacy in either the prevention or treatment of bovine mastitis. PMID- 8242459 TI - Nutrition and mastitis. AB - Increasing evidence presents a relationship between nutrition and mammary resistance to infection. The role of nutrition in mammary resistance has been best defined for antioxidants. To optimize this interaction, rations should be balanced and formulated for lactational stage. Although supplementation of nutrients with antioxidant function may enhance immune resistance against infections, therapeutic roles for nutritional management of clinical mastitis, as yet, are not well documented. The need to monitor herd antioxidant status as part of a mastitis control program is critical because (1) antioxidant supplementation improves mammary resistance to infectious disease during the most critical period for enhanced resistance, the dry and early postpartum periods; and (2) dry cows and heifers often are on a marginal plane of Se supplementation, particularly during periods of risk for mastitis and reproductive disorders. In areas where forages contain less than 0.1 ppm Se (dry matter) because of low soil availability, herds should be tested for Se status and, if found to be below optimum levels, increased dietary supplementation or sustained-release Se boluses should be considered. Similarly, vitamins A (or beta-carotene) and E status should be monitored in herds, especially confined herds receiving a large proportion of ensiled forages. PMID- 8242460 TI - Genetic improvement of mastitis through selection on somatic cell count. AB - Heredity influences both clinical mastitis and somatic cell score. Intramammary infection is the major cause of elevated somatic cell score. A nationwide program of genetic evaluation of dairy cattle for somatic cell score is being developed. Proper selection of artificial insemination sires, considering their genetic merit for both milk production and somatic cell score, will reduce the genetic increase in mastitis susceptibility that accompanies selection for high production. PMID- 8242461 TI - Mastitis in heifers. AB - Udder health is typically overlooked but critically important in heifers. Mastitis pathogens of the contagious as well as environmental groups have been implicated in mastitis in heifers. Procedures considered for control of heifer mastitis include prepartum treatment in primigravid heifers, separating preweaned calves to prevent suckling, fly control, and segregation of pregnant heifers from dry cows. Further investigation is needed before general recommendations can be made relative to prepartum treatment of primigravid heifers. An improved understanding of the epidemiology and pathogenesis of heifer mastitis will aid in expanding and improving control measures. PMID- 8242462 TI - The pasteurized milk ordinance. The practicing veterinarian's role in residue avoidance. AB - Regulatory laws increasingly affect the decision-making process of modern dairy practitioners and their clients. The purpose of this article is to explain the labyrinthine U.S. milk regulatory policies and to attempt to clarify those procedures a practitioner should follow to avoid regulatory concerns for both their clients and themselves. PMID- 8242463 TI - Antibiotic residue tests for mammary gland secretions. AB - Current control methods are not effective in eliminating various mastitis pathogens, and antimicrobial therapy introduces some risk of contaminating dairy products with antibiotic residues. Individuals involved in animal agriculture are sensitive to their responsibility to prevent antibiotic residues from entering the food chain. The ability to understand and perform the appropriate screening systems for determining the residue status of an individual animal, bulk tank, or tanker truck sample provides an opportunity to practice residue management and enhance the quality of dairy products available to the consumer. PMID- 8242464 TI - Preliminary evaluation of a new ultrasound bone densitometer. AB - A new ultrasound bone densitometer has been developed that measures ultrasonic properties of the os calcis, namely, the speed of sound (SOS), broadband ultrasound attenuation (BUA), and a proprietary factor derived from SOS and BUA, termed "stiffness." Short-term precision of ultrasound measurements was 1.4% for BUA, 0.2% for SOS, and 1.5% for stiffness in healthy women, and 1.1% for BUA, 0.1% for SOS, and 1.5% for stiffness in osteopenic women. One hundred seven women underwent measurements by ultrasound, together with dual energy X-ray absorptiometry (DXA) bone mineral density (BMD) measurements of the lumbar spine and proximal femur. Correlations between SOS, BUA, and stiffness measurements and DXA BMD measurements were all highly significant (P < 0.001) with r values varying from 0.54 to 0.67. BUA, SOS, and stiffness measurements were all significantly different between normal and osteopenic women even after adjusting for age, height, and weight (P < 0.05, P < 0.001, and P < 0.01, respectively). These results demonstrate that this ultrasound system measures ultrasonic properties of the os calcis with good precision, the measurements correlate moderately well with DXA BMD measurements and they can differentiate between normals and those with osteopenia. PMID- 8242465 TI - Measurements of postmenopausal bone loss with a new contact ultrasound system. AB - Measurements of broadband ultrasonic attenuation (BUA) and velocity of ultrasound through the heel (heel velocity, HV) were performed with a Contact Ultrasonic Bone Analyzer (CUBA-Research model) in 229 women. The subjects consisted of 16 healthy young volunteers (Group 1, mean age 26 years), 170 healthy pre- and postmenopausal women (Group 2, mean age 53 years), and 43 osteoporotic women with radiographically defined vertebral crush fracture (Group 3, mean age 66 years). Subjects in Group 1 had 10 repeated measurements in a study of short-term precision. Women in Groups 2 and 3 also had dual X-ray absorptiometry (DXA) scans to measure lumbar spine and femoral neck bone mineral density (BMD). The BUA and HV measurements for all 229 women showed a significant correlation (r = 0.75, P < 0.001). The precision study on the subjects in Group 1 gave a root mean square coefficient of variation of 6.3% for BUA and 1.04% for HV. Linear regression analysis gave the following relationship between BUA and age for the 170 normal women in Group 2: BUA = 83.6-0.86 (age 40) dB/MHz (r = -0.31, P < 0.001, SEE = 16.3 dB/MHz). The relationship between HV and age was as follows: HV = 1614-2:3 (age 40) m/s (r = -0.33, P < 0.001, SEE = 42 m/s). Multivariate regression analysis showed that in addition to age, years since the menopause was also a significant factor in determining both BUA and HV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242466 TI - Dual X-ray absorptiometry for the evaluation of bone density from the proximal femur after total hip arthroplasty: analysis protocols and reproducibility. AB - Dual X-ray absorptiometry (DXA) instruments are now able to evaluate bone mineral density (BMD) of bone surrounding metal implants. The assessment of BMD around prosthetic components could provide additional information for the follow-up of total hip arthroplasty (THA). In this study, we evaluated the potential application of DXA in the field of THA. BMD was measured in the proximal femur of both THA and THA-free sides in 14 postmenopausal women 6-18 months after THA. The explored segment was divided into seven zones as proposed by Gruen et al. [18]. The precision error of BMD measurements ranged from 1.8 to 6.8% on the THA side and from 1.1 to 2% to the THA-free side. The reduction of BMD of the THA versus the THA-free side was significant in all seven zones (P < 0.01, t-test for paired data). These results showed significant differences in BMD around femoral components of THA with respect to contralateral healthy side, and demonstrate the sensitivity of DXA for detecting these changes. PMID- 8242467 TI - Impairment of bone turnover in elderly women with hip fracture. AB - Hip fracture is one of the most severe consequences of osteoporosis affecting aged women. However, abnormalities of bone turnover responsible for bone loss in this condition have not been clearly defined. To further evaluate the bone metabolic status of women sustaining hip fracture, we have prospectively measured serum osteocalcin as a marker of bone formation and urinary excretion of pyridinoline (Pyr) and deoxypyridinoline (D-pyr) cross-links as markers of bone collagen degradation in 174 independently living women (80 +/- 8 years) within a few hours after a hip fracture. Comparison was made with 77 age-matched controls (80 +/- 5 years) and 17 premenopausal women (39 +/- 3 years). In addition 15 of the patients were followed with daily measurements during the first postoperative week. At the time of admission osteocalcin was 20% lower in the fractured women compared to the elderly controls (7.6 +/- 3.8 vs. 9.5 +/- 4.5 ng/ml, P = 0.001). Pyr and D-pyr were 36% and 40% higher, respectively (P = 0.0001), than in elderly controls and 85% and 76% higher than in premenopausal controls (P = 0.0001). Serum osteocalcin did not correlate with the cortisol level measured at the same time (r = 0.03, ns), nor with serum albumin and creatinine. Serum osteocalcin remained unchanged within 18 hours after fracture, whereafter it progressively decreased until the third postoperative day. No correlation was noted between the excretion of pyridinoline cross-links and the time elapsed from fracture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242468 TI - Pyknodysostosis: imaging and laboratory observations. AB - Pyknodysostosis is a rare form of sclerosing bone dysplasia with autosomal recessive inheritance. Affected members of two families were assessed as follows: three patients underwent densitometry measurements and bone scans; four patients underwent magnetic resonance imaging (MRI) and an immunological investigation, as well as a detailed endocrinological and biochemical laboratory review. Densitometry measurements revealed values of up to 291% of age-matched normal controls; this increased bone density was mainly in the trabecular bone and not in the cortical bone. The MRI showed the cortex to be of normal thickness, whereas the increase in trabecular bone limited the space within the medullary canal. Bone scans and single photon emission computerized tomography in three patients showed an increased uptake of [99mTc]methylene diphosphonate of up to 538% of age-matched controls, which reflected the increased bone density. Monocyte function tests demonstrated a normal phagocytic capacity, but their killing activity was impaired. Interleukin-1 secretion was also impaired, which may point to the pathogenesis of the disease, in view of its function as an osteoclast activator and its role in bone resorption. PMID- 8242469 TI - Short-term effects of organic silicon on trabecular bone in mature ovariectomized rats. AB - Silicon is known to ensure an essential role in the formation of cross-links between collagen and proteoglycans during bone growth. In this study, we have evaluated the short-term effects of a preventive treatment with silanol, a soluble organic silicon (Si), on trabecular bone in mature ovariectomized rats. Three-month-old rats were sham-operated (sham) or were ovariectomized (OVX) and treated with 10 micrograms/kg/day of 17 beta estradiol (E2), or with 0.1 mg Si/kg/day or 1.0 mg Si/kg/day of silanol for 1 month. Plasma alkaline phosphatase and osteocalcin levels were increased by 50% in OVX rats compared with sham rats and were corrected by E2 but not by silanol treatment. The trabecular bone volume measured at the tibial metaphysis was decreased by 48%, and histomorphometric indices of bone resorption and formation were increased in OVX rats compared with sham, and these parameters were corrected by E2 treatment. Treatment of OVX rats with silanol decreased the osteoclast surface by 31% and the number of osteoclasts by 20%. The mineral apposition rate, the bone formation rate, and the osteoblast surface at the tibia metaphyseal area were increased by 30% at the higher dose of silanol compared with OVX rats. In contrast, silanol treatment had no effect on the periosteal apposition rate. The reduction of the metaphyseal bone resorption and the increased bone formation induced by silanol resulted in a slight improvement of the trabecular bone volume (+14%) compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242470 TI - Molybdenum-induced changes in the epiphyseal growth plate. AB - Molybdenum (Mo), at high concentrations, induces changes in the epiphyseal growth plate through its effects on copper (Cu) metabolism but it is unclear whether or not Mo can induce changes independent of its effects on copper status. To this end, the effect of Mo on longitudinal bone growth was examined in rats. Dietary Mo was given either as ammonium heptamolybdate or as ammonium tetrathiomolybdate, the latter producing a marked Cu deficiency. There was a significant reduction in longitudinal bone growth in both groups; however, growth plate width was increased only in the Cu-deficient animals due to an increase in the width of the zone of transitional/hypertrophic chondrocytes. Both glucose 6-phosphate dehydrogenase activity and cell proliferation (assessed by bromodeoxyuridine incorporation) were markedly decreased in the proliferating zone of the growth plate in both Mo-treated groups. These changes were not apparently related to changes in circulating vitamin D metabolites or insulin-like growth factor-1. The results indicate that excess Mo impairs cell proliferation within the growth plate, whereas the effects of copper deficiency are more related to chondrocyte differentiation. Thus, Mo can induce changes in longitudinal bone growth which are distinct from those resulting from Cu deficiency. PMID- 8242471 TI - Fluoride increases net 45Ca uptake by SaOS-2 cells: The effect is phosphate dependent. AB - Previous in vitro studies have shown that the effect of fluoride to increase avian osteoblast-like cell proliferation was dependent on the phosphate concentration. In vitro studies have further revealed that fluoride could also have direct effects on osteoblast-like cells to increase phosphate uptake and transiently increase cytosolic calcium. The current studies were intended to determine whether fluoride could increase net 45Ca uptake by human osteosarcoma (SaOS-2) cells and, if so, whether those effects would also be phosphate dependent. The results of these studies indicate that fluoride increased net 45Ca uptake by SaOS-2 cells, with biphasic dose and time dependencies. After 30 minutes of exposure, net 45Ca uptake was increased to a greater extent by 50 microM fluoride (217 +/- 16% of control, P < 0.001) than by 200 microM fluoride; and the stimulatory effect of 100 microM fluoride on net 45Ca uptake was greater after 20 minutes (187 +/- 22% of control, P < 0.001) than after 60 minutes (122 +/- 7% of control, P < 0.05). These effects of fluoride to increase net 45Ca uptake were dependent on the phosphate concentration in the medium. Fluoride had no effect on net 45Ca uptake in medium containing 0.4 mM phosphate, but increased net 45Ca uptake in medium containing 1.2 or 2.0 mM phosphate (P < 0.005). As the phosphate concentration was increased, the biphasic fluoride dose-response curve was shifted to a lower range of fluoride concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242472 TI - Induction of macrophagic and granulocytic differentiation of murine bone marrow progenitor cells by 1,25-dihydroxyvitamin D3. AB - 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) was recently shown to promote maturation of 5-fluorouracil (5FU)-treated bone marrow cells by up-regulating macrophage colony stimulating factor (M-CSF) receptors in the presence of interleukin 1 alpha (IL-1 alpha). In order to reveal how 1,25(OH)2D3 interacts with colony stimulating factors and regulates the differentiation of bone marrow progenitor cell populations, in the present study, natural bone marrow cells were isolated from untreated mice and used in alpha-minimum essential medium supplemented with 20% heat-inactivated horse serum without added appropriate cytokines. Under the conditions, cells spontaneously differentiated gradually with days of culture, as assessed by expression of macrophage differentiation antigens such as Mac-1 (CD11b) and F4/80. Both M-CSF and granulocyte macrophage-colony stimulating factor (GM-CSF) induced only Mac-1 antigen expression. Simultaneous treatment with M-CSF and 1,25(OH)2D3 enhanced the M-CSF's effect on expression of both antigens, although 1,25(OH)2D3 per se has no effect on the expression for up to 11 days. In addition, successive treatment with 1,25(OH)2D3 and M-CSF or GM-CSF dramatically enhanced expression of both antigens or Mac-1 antigen, respectively. Similarly, both simultaneous and successive treatment with 1,25(OH)2D3 and M-CSF significantly enhanced phagocytic activity and H2O2 production, whereas successive treatment with 1,25(OH)2D3 and GM-CSF significantly enhanced only phagocytic activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242473 TI - Expression of insulin-like growth factors during bone induction in rat. AB - Bone formation was studied after intramuscular implantation of demineralized bone matrix. Ash weight determinations were used to verify the bone-forming ability of implants, and confirmed that no bone was formed when nonactive implants (stripped of their bone-forming ability) were used. A solution hybridization/RNase protection assay was used for the detection of specific mRNA transcripts in the implants and surrounding tissue. Analysis of insulin-like growth factor I (IGF-I) mRNA showed a transient increase peaking on day 3 following implantation. Radioimmunoassay (RIA) for IGF-I-like immunoreactivity indicated a corresponding increase of IGF-I peptide in extracts from the implants at that time point. IGF II mRNA and alkaline phosphatase mRNA reached highest levels around day 11 following implantation. Bone formation in old rats, 50 weeks of age, was associated with lower IGF-I mRNA levels 3 days after implantation compared with young animals. IGF-II mRNA levels were also affected and tended to be higher 12 days after implantation compared with young animals. These results indicate that IGFs could be paracrine or autocrine factors in the bone-forming process. During this process, IGF-I mRNA is expressed at an early stage, in correlation with the recruitment and proliferation of surrounding mesenchymal cells, whereas IGF-II mRNA is activated significantly later, correlating to the beginning of the actual calcifying process during endochondral bone formation. PMID- 8242474 TI - Inhibitory effect of ipriflavone on osteoclast-mediated bone resorption and new osteoclast formation in long-term cultures of mouse unfractionated bone cells. AB - To study the effect of ipriflavone on osteoclast-mediated bone resorption and new osteoclast formation, we used an unfractionated bone cell culture system containing mature osteoclasts from femur and tibia of newborn mice. Ipriflavone (10(-5) M) inhibited pit formation on dentin slices and caused a decrease in the number of tartrate-resistant acid phosphatase (TRAP)-positive (+) multinucleate cells (MNCs) in a 4-day culture period in which no increase in the number of TRAP(+)-MNCs was observed in the presence of 5% fetal bovine serum (FBS) and 10( 8) M 1 alpha,25-dihydroxy-vitamin D3 (1 alpha,25(OH)2D3). During the following 12 days, both the total area of the pits and the number of TRAP(+)-MNCs increased in the control. Continuous treatment with ipriflavone also inhibited the increase in pit area during this period. These effects of ipriflavone were reversible. Furthermore, the differentiation of osteoclasts was examined when preexisting TRAP(+)-MNCs were removed by incubation in the absence of 1 alpha,25(OH)2D3 for the initial 4 days in culture dishes without dentin slices. When 1 alpha,25(OH)2D3 and ipriflavone were added to the medium on the 4th day, ipriflavone inhibited new TRAP(+)-MNC formation stimulated by 1 alpha,25(OH)2D3 in a dose-dependent manner. However, pretreatment of the cells with ipriflavone before the addition of 1 alpha,25(OH)2D3 did not inhibit TRAP(+)-MNC formation. These results indicate that ipriflavone inhibits both the activation of mature osteoclasts and the formation of new osteoclasts without affecting growth of TRAP negative progenitor cells. PMID- 8242475 TI - Skeletal effects of withdrawal of estrogen and diphosphonate treatment in ovariectomized rats. AB - The study was designed to determine the skeletal effects of withdrawal of estrogen and diphosphonate treatment in the estrogen-deplete state. Groups of ovariectomized (OVX) rats were treated with vehicle alone, estrogen, or the diphosphonates etidronate or risedronate for a 180-day period. A group of sham operated control rats was treated for 180 days with vehicle alone. All treatments were then terminated, followed by sequential sacrifice of rats at 0, 35, 90, 180, and 360 days after withdrawal of treatment. The proximal tibia from each animal was processed undecalcified for quantitative bone histomorphometry. At the end of the treatment period, vehicle-treated OVX rats were characterized by cancellous osteopenia and increased bone turnover relative to vehicle-treated control rats. Treatment of OVX rats with estrogen or diphosphonates depressed bone turnover and protected against cancellous osteopenia. During the withdrawal period, OVX rats previously treated with estrogen exhibited rapid bone loss associated with increased bone turnover. The bone protective effect of the hormone in OVX rats was nearly completely lost by 90 days of withdrawal. In contrast, OVX rats maintained low levels of bone turnover and normal cancellous bone mass at 180 days of withdrawal from diphosphonate treatment. The results suggest that estrogen-deplete women who are withdrawn from estrogen replacement are at high risk for subsequent bone loss. They further suggest that widely spaced periods of intermittent diphosphonate treatment may be sufficient to prevent the development of osteopenia in postmenopausal and oophorectomized women. PMID- 8242476 TI - Cytochalasin D reduces osteoclastic bone resorption by inhibiting development of ruffled border-clear zone complex. AB - The osteoclastic cytoskeleton has been demonstrated to be composed of microfilaments. Osteoclastic multinucleated cells were suspended on dentine slices and cultured for 24 hours in the presence or absence of cytochalasin D (CD), a specific and potent inhibitor of actin filament elongation to determine the role of this cytoskeleton. Cultured cells and co-cultured dentine slices were examined ultrastructurally. Unlike those in control cultures without CD, osteoclasts in CD-treated cultures became spherical in shape and lacked microvilli on their basolateral cell surfaces. Most importantly, CD treatment induced a complete disappearance of the ruffled border-clear zone complexes in osteoclasts, which resulted in loss of osteoclast-cytoplasmic polarity. Morphometric analysis of backscattered electron micrographs of co-cultured dentine slices revealed that CD treatment strongly inhibited the formation of resorption lacunae in a dose-dependent manner. These results suggest that the cytoarchitecture, as well as the bone-resorbing function, of the osteoclast is highly regulated by the F-actin-containing microfilamentous cytoskeleton in the ruffled border-clear zone complex. PMID- 8242477 TI - Atrial and arterial baroreceptor influences on the circulatory response to acute changes in renal perfusion. AB - We have recently reported a neurally mediated reflex increase in hindlimb vascular resistance associated with an acute decrease in renal perfusion pressure in the chloralose-urethane-anesthetized rabbit. The present study was designed to investigate the influence of this reflex in the body's integrated response to circulatory disturbances by investigating the influence of carotid baroreceptor and left atrial receptors on this reflex and assessing the effect of acute changes in renal perfusion on the heart. Interaction of the renal-generated reflex with carotid baroreceptors was investigated by independent perfusion of the carotid sinus region. Responses in hindlimb perfusion pressure, at constant flow, to changes in renal perfusion were greatest with the carotid sinus perfusion pressure (CSP) low (27 +/- 4 mmHg (1 mmHg = 133.3 Pa) increase in hindlimb pressure at low CSP vs. 19 +/- 3 mmHg increase at normal CSP) and were inhibited with maximum carotid stimulation. Partial mitral obstruction, resulting in left atrial distension and atrial receptor stimulation, attenuated the hindlimb vascular response. The increase in hindlimb pressure under control conditions was 34 +/- 10 mmHg compared with 20 +/- 5 mmHg during atrial receptor stimulation. However, acute reduction of renal perfusion pressure did not result in any changes in heart rate, cardiac output, or inotropic state. It appears that both atrial and arterial baroreflexes modify the reflex change in hindlimb vascular resistance associated with acute alterations of renal perfusion. PMID- 8242478 TI - [Carnitine levels in muscle in mice with hyperammonemia: effect of treatment with sodium benzoate]. AB - The object of this study was to measure carnitine levels in skeletal muscles of experimental mice with acute, subacute, and chronic hyperammonemia, with or without sodium benzoate treatment. Normal Swiss-ICR mice in which acute hyperammonemia was created through intraperitoneal injection of ammonium acetate showed a significant decrease in free carnitine in the skeletal muscles. The same phenomenon was observed in spf mice with chronic congenital hyperammonemia caused by an X-linked deficiency of liver ornithine transcarbamylase. In spf mice with subacute hyperammonemia created through treatment with an arginine-deficient diet, free carnitine of muscles increased compared with basal levels seen in chronic hyperammonemia. The changes in carnitine metabolism during chronic and subacute hyperammonemia were accompanied by a significant increase of creatine kinase levels in the plasma, which are an index of damage to the sarcolemma. A decrease in free and esterified levels of carnitine in the muscles was observed among normal mice treated with sodium benzoate. A decrease in esterified carnitine was also noticed among spf mice given an acute treatment with sodium benzoate as well as among normal and spf mice given a prolonged treatment with sodium benzoate. Plasma creatine kinase levels were high in normal mice, after both acute and prolonged treatments. However, the levels in spf mice treated with sodium benzoate were not significantly different from those in untreated spf controls in which a significant increase in plasma creatine kinase was already caused by chronic hyperammonemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242479 TI - Characterization of a simple animal model for nonsteroidal anti-inflammatory drug induced antral ulcer. AB - Most animal models of nonsteroidal anti-inflammatory drug (NSAID) induced gastric injury are characterized by acute, superficial erosions in the corpus region, whereas the clinically significant injury in man is the deep, antral ulcer. The purpose of this study was to characterize a model of NSAID-induced antral ulceration that more closely resembles the type of damage seen in man. Rabbits received indomethacin subcutaneously every 12 h. The progression of ulcer formation was followed by killing groups of animals after one to seven doses of indomethacin. The dose dependency of ulcer formation was assessed by giving indomethacin at doses of 1 to 20 mg/kg. Healing of antral ulcers was determined by examining the stomach at various times after administering the seventh dose of indomethacin (20 mg/kg). The effects of prophylactic treatment with misoprostol or ranitidine on ulcer formation were assessed. Indomethacin administration initially produced superficial erosions in the corpus and antrum, but with time, ulcers became apparent in the antrum. The formation of these ulcers was dependent upon the number of times indomethacin was administered and the dose. Similar ulcers could be induced with a second NSAID, diclofenac. Misoprostol treatment resulted in a significant reduction in the extent of indomethacin-induced antral ulceration, but ranitidine had no effect. Antral ulcers healed progressively following cessation of indomethacin administration and were almost completely resolved by 108 h after the final dose of indomethacin. These results demonstrate that subcutaneous NSAID administration to rabbits is a simple and reproducible method for producing ulcers that bear striking macroscopic resemblance to NSAID induced antral ulcers in man.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242480 TI - Hypoxia and beta-hydroxybutyrate acutely reduce glucose extraction by the brain in anesthetized dogs. AB - Induced ketosis may acutely prolong hypoxic tolerance by altering glucose and (or) beta-hydroxybutyrate extraction by the brain. Anesthetized dogs were ventilated and instrumental for arterial and cerebral venous blood sampling, taken during baseline, infusion, test, and recovery periods. Each dog received one of four different beta-hydroxybutyrate doses and 25 min of either a normoxic or hypoxic (8% oxygen) test period. Arterial glucose and beta-hydroxybutyrate were stable in the saline and 3 mg.kg-1 x min-1 beta-hydroxybutyrate infusion groups, but glucose declined and beta-hydroxybutyrate increased in the 10 and 20 mg.kg-1 x min-1 beta-hydroxybutyrate infusion groups. During normoxia glucose arteriovenous differences across the brain decreased significantly only in the 20 mg.kg-1 x min-1 beta-hydroxybutyrate infusion groups, and during hypoxia the differences decreased for all groups. beta-Hydroxybutyrate arteriovenous differences tended to increase with a 100-fold increase in beta-hydroxybutyrate concentrations, but the trend was not statistically significant. Hypoxia further decreased glucose extraction, but did not change beta-hydroxybutyrate extraction. We conclude that beta-hydroxybutyrate may protect during hypoxia by depressing glucose uptake and consumption rather than by acting as an alternative cerebral energy substrate. PMID- 8242481 TI - Contractile responses and signal transduction of endothelin-1 in aorta and mesenteric vasculature of adult spontaneously hypertensive rats. AB - The contractile responses and generation of intracellular second messengers in response to endothelin-1 (ET-1), a potent vasoconstrictor peptide released locally by endothelial cells and involved in the regulation of vascular tone, were investigated in different segments of the vascular tree of adult 18-week-old spontaneously hypertensive rats (SHR) as compared with age-matched Wistar-Kyoto (WKY) rats. Aorta rings of SHR showed lower maximum response to ET-1 in comparison with WKY rats. Rings of the main superior mesenteric artery of SHR and WKY showed similar responses to ET-1. Small mesenteric resistance arteries of SHR, mounted on a wire myograph, developed similar tension to those of WKY rats in response to ET-1. The dose-response of inositol phosphates to ET-1 was significantly blunted in thoracic aorta of SHR compared with WKY rats, whereas it was similar in the mesenteric arterial bed. Baseline 1,2-diacylglycerol content was higher in thoracic aorta of SHR than WKY, while it was similar in the mesenteric arterial bed of the two strains. The response of 1,2-diacylglycerol to ET-1 was blunted in aorta of SHR, whereas no significant differences in diacylglycerol accumulation could be found in mesenteric vessels between SHR and WKY. In small mesenteric arteries, the dose-response to ET-1 of cytosolic free calcium, measured with the fluorescent dye Fura 2-AM, was similar in the two groups of rats. We conclude that in the aorta of 18-week-old SHR there is reduced generation of second messengers (inositol phosphates and diacylglycerol), which underlies its decreased response to ET-1. In mesenteric vessels (both proximal and distal) signal transduction is similar in SHR and WKY, and as a result contractile responses in both species are comparable. The responses to ET-1 of the arterial tree in terms of contractility and second messenger generation may reflect the adaptive processes taking place as a consequence of elevated blood pressure within the arterial wall of different segments of the vasculature of SHR. PMID- 8242482 TI - Skeletal muscle ammonia production and repeated, intense exercise in humans. AB - We investigated the impact of repeated, high-intensity exercise on NH3 metabolism using the single-leg knee extensor model. The muscle glycogen level would be lowered by the initial exercise and low glycogen may stimulate NH3 production independent of any other effects of previous exercise. Therefore a high muscle glycogen condition was included in the protocol so that the pre-exercise glycogen concentration would be at least at a normal resting level for the second exercise. The subjects (n = 6) used previous exercise and (or) diet to begin the exercise with either normal (87.0 +/- 14.4 mmol/kg wet weight) or high (176.8 +/- 22.9 mmol/kg wet weight) glycogen (C and HG, respectively) in the quadriceps. They exercised (Ex1) one leg to exhaustion (140% leg VO2 max), rested 1 h, repeated the exercise (Ex2), and then repeated the protocol with the opposite leg. The exercise durations of Ex1 and Ex2, respectively, for C were 2.82 +/- 0.51 and 2.47 +/- 0.47 min (p < 0.05) and for HG were 2.92 +/- 0.57 and 2.77 +/- 0.50 min. The NH3 efflux was reduced (p < 0.05) from Ex1 to Ex2 in both C (516 +/ 159 and 250 +/- 69 mumol, respectively) and HG (618 +/- 233 and 275 +/- 124 mumol, respectively). While NH3 efflux was virtually identical between C and HG in both Ex1 and Ex2, HG consistently had a greater arterial NH3 concentration (p < 0.05). The decreased efflux in Ex2 compared with Ex1 was not due to greater accumulation of muscle NH3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242483 TI - Active calcium absorption in primary cultures of cortical collecting duct cells. AB - Primary cultures of rabbit cortical collecting duct (CCD) cells demonstrated accumulation of Ca at the basolateral (BL) side when cultured on either impermeable or permeable supports. Cell monolayers cultured on impermeable plastic surfaces absorbed Ca with such avidity that hydroxyapatite crystals formed. When cultured on a permeable difference. A steady-state BL/A [Ca] ratio of 120 developed across monolayers in 24 h on days 6 through 8 postseeding. Initial rates of unidirectional 45Ca fluxes on days 6 through 8 indicated a negligible BL to A flux (5.4 +/- 2.6 nmol.h-1 x cm-2) compared with A to BL 45Ca flux (99.4 +/- 19.4 nmol.h-1 x cm-2). Parathyroid hormone applied to the BL side had no significant effect on either unidirectional 45Ca flux, but the second messenger analog, 8-bromoadenosine cyclic monophosphate, increased the A to BL flux by 65%. Inhibiting the Na(+)-K+ ATPase with ouabain (10(-4) M) reduced the A to BL flux by 77%; however, a significant net A to BL flux still remained. Apical addition of amiloride (2 x 10(-5) M) did not affect either unidirectional 45Ca flux. In addition, the inorganic Ca channel blockers Ni2+ (100 microM and 1 mM), La3+ (100 microM and 1 mM), and Cd2+ (20 and 50 microM) did not significantly inhibit either unidirectional 45Ca flux. These results demonstrate that CCD monolayers actively absorb Ca and this can be stimulated by cyclic AMP, raising the possibility that apical Ca entry does not involve amiloride-sensitive channels, or typical Ca channels. PMID- 8242484 TI - Neurokinin receptor mediated plasma extravasation in guinea pig and rat airways: comparison of 125I-labelled human fibrinogen and 99mTc-labelled human serum albumin as markers of leakage. AB - In the present study we have characterized NK-1 and NK-2 receptor induced microvascular leakage in guinea pig and rat airways, using 125I-labelled human fibrinogen ([125I]FN) versus 99mTc-labelled human serum albumin ([99mTc]HSA) as markers for plasma protein extravasation. Intravenous administration of the selective NK-1 agonist [Sar9, Met(O2)11]SP (1 nmol kg-1) caused a dose-dependent increase of [125I]FN extravasation in guinea pig trachea, main bronchi, secondary bronchi, and intraparenchymal airways. Extravasation of [125I]FN increased by up to 192 (trachea), 284 (main bronchi), 368 (secondary bronchi), and 271% (intraparenchymal bronchi) over control levels in these regions of the airways. Pretreatment of the animals with CP 99,994 and RP 67,580, two NK-1 nonpeptide antagonists, caused a dose-dependent inhibition of [Sar9, Met(O2)11]SP-induced leakage of [125I]FN. [Sar9, Met(O2)11]SP (1 nmol kg-1) did not induce specific leakage of [99mTc]HSA in the intraparenchymal bronchi. Specific NK-2 receptor induced leakage was detected in the lower airways but only when using [125I]FN as a marker. We have also compared the ability of CP 99,994 and RP 67,580 to inhibit [Sar9, Met(O2)11]SP induced extravasation of [125I]FN in rat airways. CP 99,994 was 40-50 (tracheobronchial region) to 75 (lower airways) times more potent in the guinea pig than the rat airways. In contrast, RP 67,580 had higher affinity for rat airways compared with guinea pig airways. The results of this study highlight the superiority of [125I]FN as a sensitive marker of plasma extravasation over [99mTc]HSA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242485 TI - Voltage-dependent calcium fluxes in skeletal muscle transverse tubule membranes in the range of late afterpotentials. AB - Calcium flux responses mediated by voltage-dependent calcium channels have been studied in transverse tubule membrane vesicles from rabbit skeletal muscle. Vesicles were loaded with 45Ca2+, and membrane potentials were generated by establishing potassium gradients across the membrane in the presence of valinomycin. After the membranes were polarized to an estimated -80 mV to approximate the resting state of the cell, a significant 45Ca2+ efflux occurred upon subsequent depolarization to -60 mV. The efflux response was modulated by activators and inhibitors of slow, dihydropyridine-sensitive calcium channels, being inhibited by inorganic calcium channel blockers, verapamil, nifedipine, and (-)-SDZ 202-791 and potentiated by the dihydropyridine agonists (+/-)-Bay K8644 and (+)-SDZ 202-791. These results demonstrate that calcium channels in transverse tubule membranes can open to mediate calcium flux in the same range of membrane potential as the late afterpotentials that occur during tetanic contractions of intact muscle fibres. PMID- 8242486 TI - Microorganisms in the gastrointestinal tract of the rat prevent absorption of the mutagen-carcinogen 3-amino-1,4-dimethyl-5H-pyrido(4,3-b)indole. AB - The extent to which lactic acid bacteria, intestinal bacteria, and yeast from the gastrointestinal tract of rats suppress the absorption of 3-amino-1,4-dimethyl-5H pyrido(4,3-b)indole (Trp-P-1) was investigated. Trp-P-1 was absorbed from the small intestine very rapidly, but in the stomach it was slowly absorbed, requiring 1 or 2 h after administration. When mixtures of Trp-P-1 and freeze dried microorganisms were administered to rats for 1 h, the amounts of Trp-P-1 absorbed from the small intestine were significantly reduced, and the levels of Trp-P-1 in blood decreased by 40.4-64.7% compared with a control in which only Trp-P-1 was administered. There were no significant differences between the organisms used. In vitro, freeze-dried cells of the strains tested bound 51-97% of Trp-P-1. The Trp-P-1 bound to cells was effectively extracted by aqueous methanol, ethanol, ammonia (50 g/L), and solutions of MgCl2 and CaCl2 (100 mM/mL), but little was extracted by water and solutions of KCl, NaCl, and buffers at various pH values. PMID- 8242487 TI - Production of a cloned xylanase in Bacillus cereus and its performance in kraft pulp prebleaching. AB - Xylanase production from a Bacillus subtilis gene cloned into a strain of Escherichia coli was monitored. Although this gene was expressed in E. coli at several temperatures, efficient xylanase secretion did not occur; the observed protein release apparently depended on cell leakage or lysis. Screening for a better microbial protein secretor free of cellulase selected Bacillus cereus No. 518. A bidirectional vector plasmid (pMK3) was employed to carry the cloned gene into this B. cereus strain. Transformation was carried out by electroporation. Total xylanase production by the new pMK3-borne gene in B. cereus was similar to that from E. coli but the xylanase was shown to be normally secreted. The xylanase gene products from the E. coli and B. cereus hosts were shown to function identically. Both xylanases improved the delignification of unbleached softwood and hardwood kraft pulps, thus reducing the Cl2 required to achieve a given degree of bleaching, without altering the physical properties of the fibers. Using a target kappa number (lignin content) of 5, xylanase pretreatment of aspen kraft (chemical) pulp led to a 22% savings of chlorine. Adsorbable organic halogens in the bleachery effluent were also lowered by more than 50%. PMID- 8242488 TI - The production of polyclonal antibodies against the mycotoxin derivative patulin hemiglutarate. AB - The mycotoxin patulin is a toxic, carcinogenic, unsaturated lactone produced by a number of molds. Polyclonal antibodies against patulin hemiglutarate were produced. Specific antibodies against patulin alone, however, were not clearly demonstrated. Because of its low molecular weight, patulin required conjugation to bovine serum albumin (BSA) to increase its immunogenicity. Anti-patulin hemiglutarate-BSA antibody titer and specificity were determined using indirect and indirect competitive ELISA, respectively. Immunoassays would facilitate detection and quantitation of patulin. PMID- 8242489 TI - MutY repair is mutagenic in mutT- strains of Escherichia coli. AB - We have determined the numbers and types of mutations that occur in strains of Escherichia coli defective in mutT and (or) mutY repair. High rates of C.G to A.T mutations in mutY- cells are unaffected by the status of mutT. However, mutT /mutY+ strains have higher rates of A.T to C.G mutations than mutT-/mutY- strains. This result indicates that the high rates of A.T to C.G mutations seen in mutT- strains of E. coli are due in part to the activity of the mutY repair system. We conclude that mutY repair is mutagenic in a mutT- background. PMID- 8242490 TI - Antibacterial activity of multilayer silver-copper surface films on catheter material. AB - The antimicrobial activity of Ag, Cu, and layered Ag-Cu surface films, sputter coated onto several types of catheter material, against clinical isolates of Staphylococcus epidermidis and Staphylococcus aureus was evaluated. When 20 microL of a suspension of S. epidermidis or S. aureus (2.68 x 10(6) colony forming units/mL) was applied onto Ag-Cu- or Cu-coated butyl rubber, bacterial numbers were greatly reduced within 10 h, and eliminated within 24 h. In contrast, antibacterial activity was significantly less on uncoated or Ag-coated surfaces. Ag-Cu- or Cu-coated silicon rubber, polyvinylchloride, and teflon were even more effective than Ag-Cu- or Cu-coated butyl rubber. Ag-Cu layered surface films also showed antibacterial activity against Pseudomonas aeruginosa biofilm formation. Multiple metal surface film combinations show great promise in lowering the incidence of device-associated nosocomial infections. PMID- 8242491 TI - Paying the extra. PMID- 8242492 TI - Paying the extra. PMID- 8242493 TI - Paying the extra. PMID- 8242494 TI - Mandatory bicycle helmets? PMID- 8242495 TI - Mandatory bicycle helmets? PMID- 8242496 TI - Mandatory bicycle helmets? PMID- 8242497 TI - Mandatory bicycle helmets? PMID- 8242498 TI - Support for future physicians. PMID- 8242499 TI - Epidural analgesia during labour. PMID- 8242500 TI - Colour-blind drivers and traffic signals. PMID- 8242501 TI - Age and driving. PMID- 8242502 TI - Aluminum and Alzheimer's disease. PMID- 8242503 TI - The ethics and economics of out-of-laboratory testing. PMID- 8242504 TI - Clinical-trial registration: a call for its implementation in Canada. PMID- 8242505 TI - Effectiveness of intrapartum penicillin prophylaxis in preventing early-onset group B streptococcal infection: results of a meta-analysis. AB - OBJECTIVE: To determine the effectiveness of intrapartum penicillin prophylaxis in preventing early-onset group B streptococcal (GBS) infection in neonates of women whose birth canals are colonized by group B streptococci. DATA SOURCES: Articles published between 1966 and 1992 identified from MEDLINE, EMBASE, the Science Citation Index and the Oxford Perinatal Database; the bibliographies of primary studies, textbooks and review articles and published abstracts from major conferences and symposia. DATA SELECTION: Studies were selected if four criteria were met: (a) the target population was intrapartum women and neonates, (b) the intervention was penicillin prophylaxis, (c) invasive early-onset GBS infection was an outcome measure, and (d) the studies were controlled trials or cohort studies. Seven primary studies were identified, four of which were randomized controlled trials. DATA EXTRACTION: Explicit methodologic criteria were used by two of the authors to assess independently the study quality; one of the reviewers was blind as to author, institution and journal. The baseline characteristics of the population, intervention and outcome were summarized twice and checked for accuracy by two of the authors. DATA SYNTHESIS: Five of the studies showed a trend toward a beneficial effect of penicillin prophylaxis, and two showed a statistically significant effect. The pooled odds ratio indicated a 30-fold reduction (95% confidence interval 0.0013 to 0.17) in the incidence of early-onset GBS infection with intrapartum penicillin prophylaxis. Subgroup analyses did not change these results. The magnitude of improvement observed did not differ between women with prenatal risk factors (premature rupture of the membranes and premature labour) and those without these risk factors. CONCLUSIONS: There is accumulative evidence that intrapartum penicillin prophylaxis is effective in preventing early-onset GBS infection. Such therapy is beneficial to women whose birth canals are colonized with group B streptococci. Further studies are needed to determine the optimum timing and method of detecting vaginal colonization during pregnancy. PMID- 8242506 TI - Illness after measles-mumps-rubella vaccination. AB - OBJECTIVES: To provide accurate information on the common sequelae of measles mumps-rubella (MMR) vaccination and to compare post-vaccine symptoms in children vaccinated at 13 and 15 months. DESIGN: Prospective cluster randomized controlled trial. SETTING: Twenty-two family practices in southwestern Ontario. PATIENTS: All 376 infants who were due to receive MMR vaccine in the next year, 253 (67.3%) successfully completed the study. INTERVENTION: MMR vaccine administered at 13 months by half of the family physicians and at 15 months by the remaining half. OUTCOME MEASURES: Family physician's physical findings in children 7 days and 30 days after vaccine; reported illnesses by mothers in a daily diary in the month before and after vaccination and medical records of visits to family physicians and hospital admissions in the month before and after vaccination. RESULTS: Compared with the incidence rates in the corresponding weeks before vaccination, the rates of lymphadenopathy (23.8%) and fever (16.8%) were higher 1 week afterward and the rate of rash (26.9%) was higher 7 to 14 days afterward. Fewer health problems were reported in the third and fourth weeks after vaccination than in the corresponding weeks beforehand. Hospital admissions after vaccination were no more frequent than those before once cause and time of admission were taken into account. The two age groups did not differ in any of the outcomes. CONCLUSIONS: Mothers should be informed about the possibility of increased physical findings in the weeks after MMR vaccination, especially lymphadenopathy, nasal discharge and rash. Since the occurrence of sequelae does not seem to differ significantly between 13-month-old recipients and 15-month-old recipients, it should not influence the decision of when to administer the vaccine. PMID- 8242507 TI - Idiopathic CD4+ T-lymphocytopenia in two patients. PMID- 8242508 TI - The genes of eye disease: the light beckons. PMID- 8242509 TI - Mandatory retirement: should older MDs be forced to retire to make way for the new? PMID- 8242510 TI - Mail-order sales of drugs without prescriptions under investigation by US, Canadian agencies. PMID- 8242511 TI - Competition doesn't do for health care what it does for the price of hamburgers. PMID- 8242512 TI - Girls, women ignoring message about dangers of tobacco, doctors warned. PMID- 8242513 TI - Degree of ill health among natives is unacceptable, CMA tells royal commission. PMID- 8242514 TI - Hospitals apply business techniques in attempt to cut cost of delivering care. PMID- 8242515 TI - Is it time for technicians to perform some surgery? PMID- 8242516 TI - Organized medicine must assume a leadership role in protecting Canada's health care system. PMID- 8242517 TI - Anorexia nervosa and depression in a woman blind since the age of nine months. AB - A woman aged 58 who has been blind since the age of nine months presented with major depression and a 40 year history of an eating disorder characterized by a restriction of food intake and body disparagement. The case is additional evidence that a specifically visual body image is not essential for the development of anorexia nervosa and supports the view that the concept of body image is unnecessary and unproductive in eating disorders. Greater emphasis should be placed on attitudes and feelings toward the body, and the possibility of an eating disorder should be considered in cases of older women with an atypical presentation. PMID- 8242518 TI - Childhood sexual abuse, parental impairment and the development of borderline personality disorder. AB - Childhood sexual abuse in borderline patients made a significant independent contribution to the magnitude of borderline characteristics after controlling for other antecedents and degree of parental impairment. This finding strengthens the etiologic link between childhood sexual abuse and borderline personality. PMID- 8242519 TI - Anxiety and depression in patients with abdominal bloating. AB - Eighteen patients with a syndrome of abdominal bloating and discomfort were examined to explore the relationship between their symptoms and their emotional problems. They were compared with 33 patients with Crohn's disease and 38 normal, healthy volunteers. Using the Hospital Anxiety and Depression Scale, patients with bloating were found to resemble patients with Crohn's disease. Both groups showed increased anxiety and depression. After controlling for age, sex, education, occupation, personality variables and childhood experience, there was a trend towards more anxiety in the bloating group compared with normal subjects and a significant difference for depression. These characteristics appear to be related to the effects of the illness or to selection, but not to personality or childhood experience. Although psychiatric problems are common among patients with abdominal bloating and pain who stay in touch with a clinic, they are not the primary cause of the disorder. PMID- 8242520 TI - The wing of madness: the illness of Vincent van Gogh. AB - This paper briefly describes some aspects of Vincent van Gogh's life and attitudes. It discusses absinthe and several psychodynamic factors that may have contributed to his psychotic episodes at Arles, when he cut off his ear. It discusses Vincent's descriptions of his illness, especially at Saint Remy de Provence and concludes that he probably suffered from partial complex seizures (temporal lobe epilepsy) with manic depressive mood swings aggravated by absinthe, brandy, nicotine and turpentine. PMID- 8242521 TI - [Panic disorder and alcoholism: effects of comorbidity]. AB - Both epidemiological and clinical studies have demonstrated a high prevalence of panic disorder among alcoholic patients. In contrast, little attention has been given to studying alcohol abuse and/or dependence in patients suffering from panic disorder. One hundred and fifty-five consecutive referrals for treatment for panic disorder were interviewed using a modified version of the Schedule for Affective Disorders and Schizophrenia--Lifetime Version, modified for the study of anxiety disorders. Thirty-two patients (20.7%) had a lifetime history of alcohol abuse and/or dependence. Although the lifetime comorbidity rate of either agoraphobia and/or social phobia seems without any influence on the risk of alcohol-related disorder, alcoholic patients suffering from panic disorder appear to be more likely to have a history of depression and other addictive disorders. The majority of patients with primary alcoholism were male, and those who became alcoholics after they developed panic disorder were more likely to be female. The comparison between patients with primary and secondary alcoholism did not indicate any difference in the comorbidity rate with other psychiatric disorders nor the severity of panic disorder. PMID- 8242522 TI - Housing for people with mental illnesses: a comparison of models and an examination of the growth of alternative housing in Canada. AB - The decrease over the past three decades in institutional care for people with mental illness has not been matched by the adequate development of specialized housing in the community. Broadly speaking, two major models of housing have emerged in the past 30 years: custodial and alternative. Large custodial settings, including boarding houses, nursing homes and special care homes, are typically not associated with residents' satisfaction or a positive outcome. Alternative housing refers to a range of models, including halfway houses, group homes, co-ops and supported housing. It is associated with better residents' outcomes. This paper briefly compares the two models and examines key issues in alternative housing and its development in Canada. PMID- 8242523 TI - Therapist-patient sexual contact: a non consensual, inherently harmful activity. AB - Therapist-patient sexual contact is unethical, whether it is the patient or the therapist who attempts to sexualize the therapeutic relationship. Therapist patient sexual contact is non consensual; patients cannot provide a morally valid consent to sexual relations with their therapist because the key elements of a morally valid consent--intentionality, substantial understanding, substantial voluntariness and autonomous authorization--cannot be met. As well, therapist patient sexual contact harms patients, patients' families, and the profession. The most significant harm caused is experienced by the patients, the majority of whom are women. PMID- 8242524 TI - [Changes by Balint group education: experience with residents in family medicine]. AB - The article is about the change observed in participants in a training course by the Balint groups. The authors' experiment is aimed at residents in family medicine. It involves both the psychiatrist and physician, leader and participant. Psychic functioning as a process is applied to presentation, discussions and leadership. The optimum clinical understanding of this training strives for development within the group itself. The authors consider two factors that come into play: symbolization and awareness. These concepts are developed to support the notion of a global approach. PMID- 8242525 TI - Cluster suicide in rural western Canada. AB - This paper focuses on a series of adolescent suicides which occurred in a small rural community in Western Canada between December 1989 and June 1990. Risk factors for adolescent suicide and recent epidemiological data on cluster suicide are reviewed and discussed. The circumstances of the five adolescent suicides are then discussed, and the question of whether or not this was a cluster suicide is considered. This article supports the view that suicide is an abnormal response to stress or loss and emphasizes the role of genetic psychophysiological predisposition. PMID- 8242526 TI - Cognitive and behavioural efficacy of clozapine in clinical trials. PMID- 8242527 TI - A comparison of mail and telephone interview strategies for mental health surveys. AB - The high cost of mental health surveys of the general population has sparked interest in less costly research methods. Two low-cost mental health survey strategies (mail and telephone) were compared in terms of cost, response rate and quality of data obtained. A total of 1,074 persons agreed to participate in the study as a sample, one-half by telephone and the other half by mail. They completed the Diagnostic Interview Schedule Self-Administered, a questionnaire designed to be self-administered, which was used to assess specific mental disorders and to evaluate risk factors. In addition, 239 respondents who were selected according to the presence or absence of specific diagnoses were reinterviewed face-to-face using the Diagnostic Interview Schedule as an external criterion. The telephone method yielded a better response rate (15% higher) and better control over answers (for example, less missing data). The mail strategy was less expensive and appeared to yield data of slightly better quality, particularly for respondents suffering from anxiety disorders. PMID- 8242528 TI - Homicide, schizophrenia and substance abuse or dependency. AB - Few studies have extensively studied the aggressive behaviours of mentally disordered offenders. This investigation compared 14 schizophrenics found not guilty of homicide by reason of insanity (NGRI) with 12 schizophrenics convicted of homicide. A comparison group of 15 homicide offenders with no major mental disorder was used. Drug and alcohol consumption, previous history of aggression against others as well as mental health were assessed using standardized, reliable, valid instruments. Significantly more of the inmates with no major mental disorder were diagnosed as having a history of drug or alcohol abuse or dependency (60%) than the NGRI schizophrenics (35.7%). In addition, both groups of convicted homicide offenders were more likely to have committed homicide under the influence of drugs or alcohol than the NGRI group. No significant difference distinguished the groups for the mean number of aggressive incidents. The subjects found NGRI assaulted more often during an acute phase of mental illness than the convicted schizophrenics. Although both groups appeared to have a similar number of hospitalizations, most of the hospitalizations of the convicted schizophrenics occurred after the crime. PMID- 8242529 TI - [Informed consent: a prerequisite for psychotherapy?]. AB - Is it desirable to extend the practice of explicit informed consent to psychotherapy? Is it possible? How? This article offers preliminary answers to these questions following a literature review and reflection on the theory of informed consent and psychotherapy. It explores the advantages and the possibility of using explicit informed consent in place of the implicit consent obtained currently. Explicit consent for psychotherapy would be interactive, iterative, repeated and revokable. It would take into account specific characteristics of psychotherapy, such as the fact that it is a process rather than a procedure, its relative unpredictability, the active part the patient plays in treatment, and the importance of the unconscious, transference and counter-transference. It would have the advantages of respecting the patients' autonomy without sacrificing their well-being, of having meaning not only for medico-legal protocols but for the patient, and would improve satisfaction with and the efficacy of psychotherapy as a treatment modality. The proposed model provides direction for future research, both empirical and theoretical. PMID- 8242530 TI - Homicide followed by suicide: a Quebec case series, 1988-1990. AB - Homicide followed by suicide is a rare but tragic event. Psychiatrists and mental health professionals are often called upon to comment publicly on the event or to help the "hidden victims," the survivors of the tragedy. There were 39 such cases in Quebec between 1988 and 1990, and they have been reviewed through the examination of the coroner's files. The epidemiological rates of homicide, suicide, homicide-suicide and the sociodemographic and clinical characteristics of the perpetrators and victims are presented and compared with the international literature. Sociological and psychopathological hypotheses are presented with a special reference to the possibility of various subgroups of perpetrators. Research needs to be pursued along the lines of more detailed case studies of psychopathological characteristics using the psychological autopsy approach. The reconstruction of the couple's relationship and of the final circumstances leading up to the tragedy may provide warning indices that could be helpful in preventing such tragedies. PMID- 8242531 TI - The child abuse reporting laws: an ethical dilemma for professionals. AB - A central issue in the development of family law and child protection in this country has been the right of the state to intervene in family life. The reporting laws, which were developed in the 1960s, made it mandatory for any citizen, including professionals, to report child abuse (physical and sexual) to the authorities. These laws have fundamentally altered the relationship between the clinician and families in need of help and have resulted in a dramatic increase in the number of cases assessed and treated by child welfare agencies. Because of the emphasis on case-finding, the limited resources have been stretched to the point where families with serious problems of child abuse do not get the services they require. Apparently, at least 60% of all reports turn out to be unfounded. Other criticisms of the current system include ineffective intervention, over-intervention and ignoring parents' rights. The legalization and bureaucratization of the child protection process has profoundly affected the relationship between troubled families and social workers who now must "investigate" them as well as help them. This role conflict is exacerbated even further in cases of alleged child sexual abuse, where social workers are often expected to inform the police of the allegations. The authors argue that the reporting laws have been useful in that society is aware of the problem, and they suggest that it may be more beneficial now to dispose of them.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242532 TI - Adrenal gland weight and suicide. AB - The purpose of this study is to determine if there is an anatomical correlate, namely adrenal hypertrophy, among people who have committed suicide. The adrenal weights and other relevant information were collected prospectively from 118 consecutive coroner's cases of sudden death in the province of Ontario. No statistically significant difference was found between the adrenal weights of those who had committed suicide, whether violent or non violent, and those dying suddenly of causes not self-inflicted. This was true irrespective of the age of the subjects, their sex or the centre at which the autopsy was performed. This finding does not support the findings of an earlier report of increased adrenal weight in successful suicides. The incidental finding of increased adrenal weight in all subjects is of some significance. PMID- 8242533 TI - A review of the amendments to the Criminal Code of Canada (Mental Disorder). AB - On February 4, 1992, Bill C-30, an Act to Amend the Criminal Code of Canada (Mental Disorder), the National Defence Act and the Young Offenders Act, was officially proclaimed. The authors provide an overview of this legislation and raise some concerns regarding certain sections of the Criminal Code. PMID- 8242534 TI - Long term strategies for reducing the shortage of psychiatrists. PMID- 8242535 TI - Post-abortion survivors syndrome. PMID- 8242536 TI - Is simple schizophrenia a variant of negative syndrome? PMID- 8242537 TI - A tribute to Lauren V. Ackerman. PMID- 8242538 TI - Cure for early gastric cancer. PMID- 8242539 TI - Adrenal cortical carcinoma. Epidemiology and treatment with mitotane and a review of the literature. AB - BACKGROUND: Adrenal cortical carcinoma is rare; the authors have treated only eight patients with the disease at Good Samaritan Regional Medical Center since 1974. No exhaustive collection of cases of this cancer has been done since 1952. METHODS: The authors retrospectively reviewed the medical records of their eight patients with adrenal cortical carcinoma. They also searched the English literature from 1952 to 1992 for reports of patients with the disease. They treated each report as a series if two or more previously unreported patients were reported. They paid special attention to patients for whom stage of disease was noted at diagnosis, treatment with mitotane (o,p'-DDD) was used, and the outcome was reported. RESULTS: Five were male and three were female patients. Five had nonfunctional tumors. None were pediatric. The authors found 1891 cases in the English literature. Adrenal cortical carcinomas are more common in women (58.6%) than in men (41.4%). The age distribution of tumors is bimodal, with peaks in the first and fifth decades. Tumors in children are more commonly functional (83.5% in female patients, 85.6% in male patients), although nonfunctional tumors are more common in older patients (84.7%). Most (68%) of these tumors are diagnosed late in disease when surgery is no longer curative. Only 35% of patients treated with mitotane had a clinical response. CONCLUSIONS: Adrenal cortical carcinomas are diagnosed most often in children because of functionality and older men because of mass effect. Most tumors are discovered too late for curative resection. Treatment of metastatic disease with mitotane has limited success. PMID- 8242540 TI - Recurrence of early gastric cancer. Follow-up of 1475 patients and review of the Japanese literature. AB - BACKGROUND: The outcome of surgical treatment for early gastric cancer (EGC) generally is considered to be satisfactory. Although a small percentage of patients experience disease recurrence, the absolute number of such instances is too small for assessment of risk factors. METHODS: The authors investigated the follow-up records of 1475 patients with EGC treated at the National Cancer Center, Tokyo, with special reference to cancer recurrence. In addition, 20 Japanese reports on this subject were reviewed. RESULTS: Excluding operative deaths and patients with noncurative operations, 20 (1.4%) patients died of recurrent disease. The death rate associated with other causes (6.6%), including other malignant diseases, surpassed that associated with disease recurrence. Late recurrence (after 5 years) was seen in seven patients. By combining the data of patients reported in 20 articles in the literature, the authors estimated a recurrence rate for EGC of at least 1.9%, exclusive of cancers arising in the gastric stump. The incidence of recurrence was significantly higher in submucosal (3.6%), node-positive (10.7%), and histologically differentiated carcinomas (2.3%) than in mucosal, node-negative, and undifferentiated groups, respectively. Analysis of 123 instances of recurrent disease, the details of which had been reported, revealed that hematogenous metastasis was the most common mode of recurrence. The mean survival period of patients with recurrent disease was 40 months, and 23% of patients died more than 5 years after surgery. CONCLUSION: It is rare for EGC to recur, even in patients with tumors with a comparatively high risk of recurrence. Long-term follow-up of patients with positive nodes for hematogenous spread might facilitate early detection of disease recurrence. PMID- 8242541 TI - 5-fluorouracil and interferon-alpha-2a in advanced colorectal cancer. Results of two treatment schedules. AB - BACKGROUND: Potential synergy between 5-fluorouracil (5-FU) and interferon alpha 2a (IFN-alpha-2a) has been demonstrated in the treatment of colorectal carcinoma. Continuous low-dose infusion of 5-FU may have superior response rates to bolus 5 FU in these malignancies. This report presents results of two Phase II trials using these principles in colorectal cancer. METHODS: Forty-eight patients were entered onto two protocols; 18 were treated with 5-FU by a bolus infusion schedule with concurrent IFN-alpha-2a (Group 1). Thirty patients were treated with continuous low-dose 5-FU and IFN-alpha-2a thrice weekly (Group 2). RESULTS: The overall response rates were 33% (95% confidence interval [CI], 16-68%) and 33% (95% CI, 17-53%), respectively, for Groups 1 and 2. In Group 2, in 16 previously untreated patients, there was a response rate of 56% (95% CI, 30-80%). The median survival was 11 months and 6 months for Groups 1 and 2, respectively. Toxicity in Group 1 was as expected, except the incidence of central nervous system toxicity was low, with only one patient requiring dose reduction because of cerebellar ataxia. The toxicity in Group 2 was substantial, with four patients being removed from study because of toxicity and all patients treated for more than 2 months requiring dose reductions. The most common (67%) toxicity was mucositis, with 33% of those patients classified as Grade III or IV (Southwest Oncology Group criteria). Other major toxicities were fatigue and hand/foot syndrome. CONCLUSIONS: The first trial confirms previous response rate data for bolus injection 5-FU and IFN-alpha-2a. The second trial of low-dose continuous infusion 5-FU with IFN-alpha-2a demonstrates similar efficacy with substantially greater toxicity. PMID- 8242542 TI - A phase II study of mitoxantrone combined with beta-interferon in unresectable hepatocellular carcinoma. AB - BACKGROUND: Chemoimmunotherapy is being evaluated in the most common gastrointestinal tumors, but little data are available on hepatocellular carcinoma (HCC). Considering the encouraging objective response rates and the absence of important side effects obtained with mitoxantrone in HCC, we tested the activity and feasibility of a schedule combining beta-interferon (beta-IFN) and mitoxantrone. METHODS: Forty patients (ECOG Performance Status 0-1) with unresectable HCC received mitoxantrone (12 mg/m2 intravenously every 3 weeks) plus beta-IFN (3 x 10(6) U on days 1, 2, and 3; 6 x 10(6) from day 4 to day 60; and then 6 x 10(6) U three times a week for 10 months). RESULTS: Thirty-eight patients were evaluable for response and toxicity with a median of four administered cycles (range, 2-10 cycles). Nine patients achieved a partial response (23%) (95% confidence interval, 11-40%) with a median duration of response of 4 months. In 15 cases, the disease was stable for at least 2 months; 14 patients had disease progression. The median survival time of the group as a whole was 8 months. Patients who were alpha-fetoprotein positive had a median survival time of 7 months; those who were alpha-fetoprotein negative had a median survival time of 9 months. The most common side effects were hematologic (World Health Organization Grade 3, 15 patients; Grade 4, 3 patients). Mild or moderate flu-like syndrome was present in 50% of treated patients, whereas 10 patients experienced mild or moderate nausea. CONCLUSIONS: The schedule was active on advanced tumors with high alpha-fetoprotein values, and side effects were manageable. However, the addition of beta-IFN did not seem to improve significantly the response rate in HCC. PMID- 8242543 TI - An experimental study and clinical pilot trials on yttrium-90 glass microspheres through the hepatic artery for treatment of primary liver cancer. AB - BACKGROUND: Yttrium-90 (90-Y) glass microsphere is a new kind of radiation microsphere for internal radiation therapy of primary liver cancer (PLC). The study was carried out by administration of 35 microns nondegradable 90-Y glass microsphere through the hepatic artery for treatment of PLC. METHODS: Six rabbits were injected with 185-1480 megabecquerels (MBq) of 90-Y glass microspheres, and three rabbits were injected with 35-300 mg of 89-Y glass microspheres for the toxic test. Eighteen patients received 2442-5550 MBq of 90-Y glass microspheres for the treatment of PLC. Whole blood counts, liver function, and imaging examination were performed. Pathologic examinations were performed on all rabbits. RESULTS: All rabbits were apparently well after absorbing 114.1-845.2 Gy 90-Y glass microspheres in the liver but showed transient degeneration of hepatocytes and portal fibrosis histologically. The mean absorbed dose in liver tissue of patients with PLC was 30.33 Gy, whereas that in tumor tissue was 88 Gy and the highest in tumor tissue, 186.36 Gy. The mean tumor:liver tissue ratio was 3:1; the highest, 14:1. Fourteen patients were still alive after half a year's follow-up and 6 of these 14 were still alive after 1 year. CONCLUSIONS: The rabbits could tolerate up to eight-fold of the upper limit of clinical dose (100 Gy). Good responses to the radiation therapy of 90-Y glass microspheres in patients with localized and hypervascular or vascular mass were achieved. Contraindications for the therapy were presence of massive hepatic arterioportal shunt and cancer emboli in the main portal vein. It is safe and applicable to deliver 90-Y glass microspheres in large doses through the hepatic artery for internal radiation treatment of PLC. PMID- 8242544 TI - Clinical significance of DNA measurements in small cell lung cancer. AB - BACKGROUND: Little is known about the relationship between DNA histogram data and the clinical course of small cell lung cancer (SCLC). METHODS: The ability of tumor nuclear DNA histograms as measured by cytofluorometry to predict chemotherapeutic responsiveness and propensity for metastases was assessed in 36 patients with SCLC. Histograms were classified into four types: A, mono-mode and euploid; B, mono-mode but not euploid (A + B being monoclonal patterns); C, poly mode; and D, no mode (C + D being polyclonal patterns). The grade was classified into low (DNA content, smaller than 8C) and high (greater than 8C) by dispersion degrees about each type. RESULTS: The relative rate of response to chemotherapy was A (100%) is greater than B (82%) is greater than C (38%) is greater than D (17%) (A and D, P < 0.05; B and D, P < 0.01; A + B and C + D, P < 0.01) and closely related to type. Metastasis rates were significantly different between high (81%) and low (7%) grade (P < 0.01) and closely related to grade (sensitivity, 81%; specificity, 93%). Peripheral SCLC had a significantly lower response rate (20%) than did proximal SCLC (65%) (P < 0.01). In DNA histograms, the proportion of C + D (more heterogeneous than A+B) was significantly higher (80%) in peripheral SCLC (P < 0.01). CONCLUSIONS: Nuclear DNA measurements may be potentially useful for predicting distant metastases and response to chemotherapy. The heterogeneity and possibility of resistance to chemotherapy of peripheral SCLC may tend to be higher than those of proximal SCLC. PMID- 8242545 TI - Localized invasive pulmonary aspergillosis in patients with neutropenia. Effectiveness of surgical resection. AB - BACKGROUND: Invasive pulmonary aspergillosis (IPA) is a major cause of morbidity and mortality in patients with neutropenia. Two severe complications with poor outcome can be observed after apparently successful IPA medical treatment: severe hemoptysis and IPA relapse during subsequent cytotoxic treatments. Early surgical therapy has not been considered routinely in the management of localized IPA. METHODS: Six consecutive patients (four women, two men; median age, 52 years) with localized cavitating IPA diagnosed during chemotherapy-induced aplasia were treated with early surgical resection after hematologic recovery. RESULTS: All patients received a lobectomy. Surgery was uneventful. This procedures allows patients to proceed with further intensive chemotherapy and/or bone marrow transplantation without IPA reactivation. CONCLUSIONS: For selected patients, surgical resection of localized IPA with unique cavitating lesion, which prevents hemoptysis and IPA recurrence and allows for subsequent cytotoxic treatment, may be recommended. PMID- 8242546 TI - Primary chemotherapy and delayed surgery for nonmetastatic osteosarcoma of the extremities. Results in 164 patients preoperatively treated with high doses of methotrexate followed by cisplatin and doxorubicin. AB - BACKGROUND: Neoadjuvant chemotherapy is the most accepted treatment for localized osteosarcoma. This has led to a great improvement in limb-sparing surgery and in disease-free survival. Patients with a good response to preoperative chemotherapy showed a higher disease-free survival rate. Current studies examine the possibility of patients whose limbs could be rescued with a poor necrosis and a reduction of the side effects related to aggressive treatments. METHODS: Between September 1986 and December 1989, 164 patients entered the second neoadjuvant study conducted at the Rizzoli Institute, Bologna, Italy, for non-metastatic osteosarcoma of the extremities. Preoperative chemotherapy consisted of two cycles of high-dose methotrexate intravenously (i.v.) followed by cisplatin intraarterially and doxorubicin i.v. After surgery, patients classified as good responders (> 90% tumor necrosis) received three more cycles of these drugs, whereas poor responders (< 90% tumor necrosis) had more chemotherapy, which included ifosfamide and etoposide in addition to the other three drugs. RESULTS: Limb salvage was performed in 83% of cases. At an average follow-up of 54 months (36-76), 109 patients (66%) were continuously disease-free, 2 died from doxorubicin cardiotoxicity, and 52 experienced metastases and 3 had local recurrence. In two of these three patients, metastases followed local recurrence. The 5-year actuarial continuously disease-free survival rate was 63%, with no differences between good and poor responders. Excluding 20 patients who had major protocol violations, the projected continuous disease-free survival rate was 71%. CONCLUSIONS: With an aggressive neoadjuvant chemotherapy, it is possible to cure more than 60% of nonmetastatic osteosarcoma of the extremities, avoiding amputation in most cases. Ifosfamide and etoposide seem to be effective in patients who did not respond to preoperative chemotherapy. PMID- 8242547 TI - Familial cutaneous melanoma and two-mutational-event modeling. AB - BACKGROUND: According to the Knudson two-mutational-event theory, two mutations at a genetic locus may be required for the development of some cancers. Persons who have inherited a defect in one chromosome and therefore require only one more mutation for cancer development are at a higher risk of manifesting cancer at a younger age than persons without an inherited mutation, who need two acquired "hits." This difference allows one to distinguish familial and sporadic types of the same malignancy by evaluating age of disease onset. METHODS: To study the role of inheritance in the etiology of familial cutaneous melanoma, characteristics of patients with familial versus nonfamilial melanoma were analyzed according to the Knudson two-mutational-event model. RESULTS: The familial versus nonfamilial graphs, based on age of diagnosis, did not support this model. However, there was a statistically significant earlier age of diagnosis for patients with familial melanoma. Melanoma thickness was less (i.e., earlier cancer at possibly younger age) for patients with a positive versus a negative family history. Conversely, linear regression, after adjusting for tumor thickness, showed that patients with hereditary melanoma still manifested earlier ages of diagnosis of melanoma compared with sporadic patients. CONCLUSIONS: Genetic patterns other than the two-step model, additional family-related factors, patient-physician sensitization due to a family history, or a combination of these factors might explain this age difference. More complex multistep modeling of the data may be helpful in better characterizing the genetic patterns of cutaneous melanoma. PMID- 8242548 TI - Combination chemotherapy in adult desmoid tumors. AB - BACKGROUND: Desmoid tumors are locally aggressive tumors, with no metastatic potential, that generally are amenable to local treatments, such as surgery and radiation therapy. Systemic therapy is considered for selected cases that are not amenable to local treatment. METHODS: The authors reviewed their experience with chemotherapy in desmoid tumors. A patient population was identified through a search of the data base maintained by the Department of Patient Studies. RESULTS: Between January 1971 and December 1991, 180 patients with a histologically confirmed diagnosis of desmoid tumor were seen at the authors' institution. Twelve patients (8 male and 4 female patients; age range, 16-66 years; median age, 29 years) received chemotherapy. Eleven patients received doxorubicin (60-90 mg/m2) plus dacarbazine (750-1000 mg/m2)-based regimens for a median of 5 cycles (2-10 cycles). Six of the nine patients who could be evaluated for response had an objective response (two complete responses and four partial responses), one patient had a minor response, and two patients had stable disease. Two other patients treated in the early 1970s could not be evaluated objectively because of lack of modern imaging; however, they were reported to have "responses" that enabled resection of axillary and pelvic disease. All four patients with Gardner syndrome experienced disease response. One of these four patients had a complete response twice with doxorubicin-based chemotherapy and eventually died with an ejection fraction of 0.22. Five patients are alive with no evidence of disease (NED), four are alive with disease, and two are lost to follow-up after having an NED status at their last visit. CONCLUSION: The authors conclude that desmoid tumors in adults are responsive to chemotherapy, and such treatment should be considered before embarking on radical treatment to avoid obvious functional consequences and delayed complications. PMID- 8242549 TI - The psychological morbidity of breast cancer-related arm swelling. Psychological morbidity of lymphoedema. AB - BACKGROUND: The psychological morbidity, functional impairment, and disturbance in psychosocial adjustment to illness was evaluated in relation to breast cancer related arm swelling. METHODS: Fifty women with breast cancer-related arm swelling were matched with 50 control subjects for age, duration since treatment, and type of treatment received. All study participants were free from active disease and had been treated more than 1 year ago. RESULTS: Patients with arm swelling showed greater psychological morbidity at formal psychiatric interview, impaired adjustment to illness as evaluated by the Psychosocial Adjustment to Illness Scale, and greater impairment of physical functioning. CONCLUSIONS: Patients with arm swelling in relation to breast cancer experienced functional impairment, psychosocial maladjustment, and increased psychological morbidity. These findings have implications for management of breast cancer. PMID- 8242550 TI - Ovarian dysplasia: nuclear texture analysis. AB - BACKGROUND: Ovarian dysplasia has been defined by histologic and morphometric studies focusing on architectural and nuclear profile changes. A new technique is used to enhance the accuracy of this diagnosis by a quantitative evaluation of the nuclear texture that represents the nuclear chromatin pattern on which conventional diagnoses of malignancy are usually made. METHODS: Histologic sections from 35 ovaries classified as malignant (12), dysplastic (12), and normal (11) were evaluated by tracing boundaries of nuclear profiles and measuring "textons" (texture primitives) with a histogram analysis of three zones of gray level densities (called for simplification white, gray, and dark). The average combined area was tabulated for specimens with the same diagnosis, and linear regression plots compared the texton area with total nuclear area. RESULTS: The dimensions of textons originally hidden inside the chromatin and revealed by histograms were found to be closely clustered in normal epithelium, and increasingly dissociated from the containing nucleus as the lesion progressed from dysplastic to malignant. The statistical multivariate analysis including nine parameters correctly classified the three diagnostic categories as normal, dysplastic, and malignant. CONCLUSIONS: Computerized image analysis of nuclear texture adds accuracy to the recently elaborated morphometric methods to define ovarian dysplasia, a potential precursor of ovarian carcinoma. PMID- 8242551 TI - Combined analysis of flow cytometry and morphometry of ovarian granulosa cell tumor. AB - BACKGROUND: It is difficult to determine the prognosis of granulosa cell tumors (GCT) at the time of diagnosis. METHODS: The nuclear DNA content of 17 patients with ovarian GCT was investigated by flow cytometry using paraffin-embedded tissue. Nuclear area (NA), nuclear perimeter (NP), and nuclear shape factor (NSF) were measured by an image analyzer using hematoxylin- and-eosin-stained sections. RESULTS: The follow-up period of the patients ranged from 2 months to 11 years. Thirteen tumors were diploid or near diploid, whereas one was tetraploid, and three were aneuploid. Two tumors had varying degrees of DNA content heterogeneity. Crude survival of the patients with an euploid tumor (13 diploid, 1 tetraploid) was more favorable than that of the patients with an aneuploid tumor. Patients with S-phase fraction (SPF) greater than 10% or DNA content heterogeneity experienced disease recurrence or metastasis. A significant difference was observed in NA and NP between those with and without metastasis. CONCLUSIONS: Our results indicate that DNA aneuploidy, large SPF, DNA content heterogeneity, and large NA and NP are adverse prognostic factors in GCT. Thus, flow cytometric and morphometric measurement may provide a rapid and valuable method to predict the biologic behavior of GCT. PMID- 8242552 TI - Antagonists of bombesin/gastrin-releasing peptides as adjuncts to agonists of luteinizing hormone-releasing hormone in the treatment of experimental prostate cancer. AB - BACKGROUND: Palliative methods for treatment of advanced prostatic carcinoma, including those based on luteinizing hormone-releasing hormone (LH-RH) agonists, cannot prevent the ultimate growth of hormone-independent cells, and the duration of disease remission in patients with prostate cancer is limited. New therapeutic approaches combining androgen ablation therapy with other compounds must be explored. Various studies suggest that bombesin or gastrin-releasing peptide (GRP) act as autocrine growth factors and may play a role in the initiation and progression of some cancers, including those of the prostate. METHODS: The effects of treatment with bombesin/gastrin-releasing peptide (GRP) receptor antagonist [D-Tpi6, Leu13 psi(CH2NH)Leu14]BN(6-14)(RC-3095), an agonist of LH-RH [D-Lys6]-LH-RH and their combination were investigated in the androgen-dependent Dunning R-3327H rat prostate cancer model. Both analogs were administered by continuous subcutaneous infusion from osmotic minipumps for 7 weeks. RESULTS: Tumor volumes and weights were significantly reduced by treatment with RC-3095, compared with those of controls. In rats that received [D-Lys6]-LH-RH, there was a greater decrease in tumor weight and volume than that produced by RC-3095, and the weights of testes, ventral prostate, and seminal vesicles also were reduced. The combination of RC-3095 and [D-Lys6]-LH-RH had the greatest inhibitory effect on tumor growth. Histologic parameters demonstrated a significant increase of the ratio of apoptotic to mitotic indices in the groups treated with [D-Lys6]-LH-RH or the combination. Serum LH and testosterone levels were greatly depressed by [D Lys6]-LH-RH or the combination. Specific high-affinity binding sites for bombesin/GRP, epidermal growth factor (EGF), and insulin-like growth Factor I (IGF-I) were found on the tumor membranes. The concentration of receptors for EGF was significantly reduced by treatment with the bombesin/GRP antagonist RC-3095. CONCLUSIONS: Combination therapy of LH-RH analogs with bombesin antagonists such as RC-3095 might be considered for improvement of hormonal therapy of prostate cancer. PMID- 8242553 TI - Neuroblastoma as a prominent component of a mixed germ cell tumor of testis. AB - BACKGROUND: Primitive neuroectodermal tissue in teratomas of testis has been reported in the literature. A mixed germ cell tumor of testis with a prominent neuroblastoma component dictating the clinical behavior was found to be unique. METHODS: Tissue sections were stained with hematoxylin and eosin, and immunohistochemical, ultrastructural, cytogenetic, and flow cytometric analyses were performed on the primitive neuroectodermal component of the testicular mass. Follow-up results at 2.5 years are included. RESULTS: The microscopic findings on hematoxylin and eosin slides showed cells composing the majority of the neoplasm to have features of neuroblastoma. The immunohistochemical stains showed positivity for neuron-specific enolase in the cells comprising the neuroblastoma, and transmission electron microscopic study corroborated these findings by demonstrating microtubules and rare membrane-limited, dense-core granules in the cytoplasm. Flow cytometry showed a hypertetraploid population with a large aneuploid DNA content. Cytogenetics revealed a hypertriploid modal number of 74 chromosomes. The clinical features were dictated by the neuroblastoma component in a fashion similar to that of adult neuroblastomas and responded to the chemotherapeutic regimen designed for treating neuroblastoma. CONCLUSIONS: The neuroblastoma component proved to be more aggressive than the other elements of this neoplasm. This finding suggests that mixed germ cell tumors showing a large neuroblastoma component should be treated promptly and aggressively with chemotherapy. PMID- 8242554 TI - Primary cerebral fibrosarcomas. Clinicopathologic study and review of the literature. AB - BACKGROUND: Primary of the brain and meninges are uncommon tumors. Information regarding optimum treatment is limited due to their rarity, and the best form of therapy is not yet known. METHODS: Nine patients between the ages of 22 and 61 years with primary fibrosarcomas confined to the brain and meninges were studied clinicopathologically. Tumors were superficially located in five patients and intracerebrally or deep in four patients. Treatment consisted of maximum feasible surgical resection (seven patients, gross total; one, subtotal; one, no surgery before death) and radiation therapy (45-60 Gy conventional fractionation) in eight of nine patients. RESULTS: Fibrosarcomas were moderate or high grade in seven of nine patients (78%). Immunohistochemical reactions with antisera to glial fibrillary acid protein (GFAP), cytokeratin, vimentin, desmin, and S-100 demonstrated vimentin positivity in five of nine patients and cytokeratin positivity in two of nine patients. Individual GFAP-positive cells were seen in two cases, thought to represent trapped reactive astrocytes. None were S-100 positive or desmin-positive. Eight patients have died, with a median survival time of 7.5 months (range, 1 day-96 months). Local recurrence developed in eight patients and distant recurrence in six patients. Systemic metastases developed in four patients (50%) and meningeal seeding in four patients (50%). Longer survival was observed in superficially located tumors (range, 7.5 months-96 months) compared with intracerebral tumors (range, 1 day-9 months). CONCLUSIONS: Primary fibrosarcomas of the brain are uncommon tumors, usually of high histologic grade, with a high rate of local recurrence. Their propensity for meningeal and distant relapse distinguishes them from tumors of glial origin. Immunohistochemistry is of limited diagnostic value, although it may facilitate exclusion of other diagnoses. Because the prognosis after conventional surgery with 50-60 Gy external beam radiation therapy is relatively poor, the authors recommend more aggressive therapy with maximal feasible resection followed by external beam radiation therapy to doses of 64-66 Gy. When effective chemotherapy is established for soft tissue sarcomas of the extremities, this should be evaluated in view of the high incidence of distant metastases. PMID- 8242555 TI - Gene expression of insulin-like growth factor II in human intracranial meningioma. AB - BACKGROUND: Insulin-like growth factor II (IGF-II) is synthesized in the normal brain of adult humans predominantly in the choroid plexus and meninges and is secreted in the cerebrospinal fluid. The authors measured IGF-II transcripts and peptides in biopsy specimens from human intracranial tumors including astrocytomas, glioblastomas, and meningiomas. METHODS: The presence of IGF-II mRNA was analyzed in 12 human brain tumors by Northern analysis of total RNA extracted from tumor biopsies and by in situ hybridization of tissue sections. The amount of immunoreactive IGF-II was determined by radioimmunoassay of tumor extracts. RESULTS: Northern analysis of RNA from four meningiomas showed IGF-II mRNA of 6.0, 4.8, and 2.2 kb, and in situ hybridization revealed that meningioma tumor cells contained IGF-II mRNA. In contrast, biopsy specimens from four astrocytomas, one oligoastrocytoma, and four glioblastomas showed no IGF-II mRNA. Radioimmunoassay of IGF-II in tumor extracts showed that all tumors contained IGF II (40-160 ng/g tissue). Two meningiomas contained the highest amounts of IGF-II (144 and 160 ng/g tissue). CONCLUSIONS: IGF-II mRNA is present in higher amounts in benign meningiomas than in malignant glioblastomas and astrocytomas, whereas the content of immunoreactive IGF-II is similar. On the basis of these findings, the authors believe that IGF-II may be involved in growth regulation of meningiomas. PMID- 8242556 TI - Immunohistochemical study of adrenocortical carcinoma. Predictive value of the D11 monoclonal antibody. AB - BACKGROUND: The diagnosis of adrenocortical carcinoma (ACC) may be difficult with conventional light microscopy, especially when the tumor is nonfunctioning. Until now, no specific adrenocortical tumor marker was available. The current study was undertaken to investigate the interest of the D11 MoAb for the diagnosis and prognosis of ACC. METHODS: Eighteen adrenocortical carcinomas, 10 primary adrenomedullary tumors, 20 primary hepatocellular carcinomas, 50 primary renal cell carcinomas, 5 primary lung carcinomas, and 18 intraadrenal metastases were analyzed immunohistochemically with the D11 monoclonal antibody. ACC were also evaluated for the expression of other tumor markers, including neuron-specific enolase, chromogranin A, S-100, Leu-7, vimentin, KL1, AE1AE3, and epithelial membrane antigen. Relationships between clinical features and results of immunohistochemistry were also sought. RESULTS: Nuclear D11 staining appears to be highly specific for normal adrenocortical cells and related tumors. Nuclear D11 positivity was demonstrated in 44% of ACC and was restricted to well differentiated tumors. No cytoplasmic or nuclear D11 staining was observed in adrenomedullary tumors. D11 reactivity confined to the cytoplasm was found in 5 of 18 adrenal metastases, in all 20 hepatocellular carcinomas tested, in 3 of 5 lung carcinomas, and in 1 of 50 primary renal cell carcinomas. Patients with nuclear D11 immunostaining were initially seen with metastases less often and survived longer than those with no nuclear D11 immunostaining (P < 0.05). CONCLUSIONS: Nuclear D11 immunoreactivity may help to differentiate ACC from intraadrenal metastases and adrenomedullary tumors. This also selects a group of ACC patients with a more favorable outcome. PMID- 8242557 TI - Determinants of subsequent home health care nursing service use by hospitalized patients with cancer. AB - BACKGROUND: The authors examined the extent to which specific patient characteristics and length of hospital stay were capable of independently explaining the use of home health care nursing services by hospitalized patients with cancer after discharge. METHODS: The current study represents a secondary analysis of a data set originally gathered to identify the home health care needs of patients with cancer. The sample involved 87 patients with cancer who received home health care after hospitalization and 43 patients who did not receive such services. RESULTS: A logistic regression analysis indicated that home health care use was related to patient age, length of hospital stay, and level of symptom distress. Specifically, the likelihood of home health care use was found to increase among subjects older than 50 years of age, subjects with hospital stays of more than 7 days (apparently related to surgery), and those who experienced moderate to high levels of symptom distress. CONCLUSIONS: The results indicate a need for home health care nurses to be skilled in the management of cancer symptoms and in the complex problems commonly experienced by the postsurgical patient with cancer. PMID- 8242558 TI - Suramin for germ cell tumors. In vitro growth inhibition and results of a phase II trial. AB - BACKGROUND: Several studies have indicated a role for specific growth factors in the differentiation or deregulated growth of germ cell cancers. One proposed mechanism of antitumor activity for suramin is the inhibition of these tumor derived growth factors. METHODS: Based on these data, the effects of suramin were studied on the growth of a monolayer culture of two human teratocarcinoma cell lines, NT2/D1 and N2102ep. These represented one cell line that was sensitive (NT2/D1) and one that was resistant (N2102ep) to the differentiation agent retinoic acid. These studies were followed by a Phase II trial of suramin in patients with germ cell tumors. Patients were treated with a continuous infusion of suramin. Dosing was determined by a nomogram based on weekly serum concentrations. RESULTS: Suramin inhibited the growth of both cell types in a dose-dependent fashion at a concentration attainable in patient plasma. This provided the rationale for a clinical trial of suramin in patients with cisplatin refractory germ cell tumors. Fourteen patients were treated, and no complete or partial responses were observed. One patient achieved a prolonged decline in serum alpha-fetoprotein level without change in the size of a bidimensionally measurable retroperitoneal mass for more than 11 months. CONCLUSIONS: Despite the results of the in vitro studies, a Phase II trial failed to demonstrate significant antitumor activity for suramin in patients with cisplatin-refractory germ cell tumors. Although the dosing of suramin by the nomogram was not associated with neurotoxicity, the time needed to achieve therapeutic levels was lengthy, and the levels, once achieved, were of short duration. Thus, an alternative means of administration, i.e., by intermittent bolus administration, is being investigated in trials for patients with prostatic carcinoma. The lack of definitive efficacy for suramin in this trial has led the authors to consider other agents in Phase II trials for cisplatin-refractory germ cell tumor. PMID- 8242559 TI - Acquired angioedema as the presenting feature of lymphoproliferative disorders of mature B-lymphocytes. AB - BACKGROUND: Acquired complement component 1 (C1) inhibitor deficiency with consequent angioedema is a rare condition that may indicate an underlying neoplasm of B-lymphocytes or plasma cells. METHODS: Three patients with C1 inhibitor deficiency and late-onset angioedema were studied to establish the presence and to characterize the nature of any underlying lymphoproliferative disorder. RESULTS: Each of the patients was found to have a low-grade B-lineage lymphoma. In one patient, the underlying disorder, best characterized as splenic lymphoma with villous lymphocytes, was occult initially. The other two patients had splenic lymphoma with villous lymphocytes and small lymphocytic lymphoma, respectively. Two of the patients had an immunoglobulin M (IgM) serum paraprotein with cold agglutinin activity; the third patient had no paraprotein, but his cells expressed monoclonal membrane IgM. Two of the patients initially did not require specific anti-lymphoma therapy and responded to therapy with an attenuated androgen; in the third patient, splenectomy and cytotoxic chemotherapy led to resolution of the acquired angioedema. In all three patient, there was considerable delay in diagnosis consequent to lack of awareness of this syndrome. CONCLUSIONS: The presence of a neoplasm of B-lymphocytes was established eventually in all three patients, although in one patient it was occult initially. In each case, the lymphocytes were mature B-lymphocytes that either showed cytologic evidence of plasma cell differentiation or secreted a paraprotein. Underlying lymphoma is common in patients who are seen initially with late-onset angioedema. Angioedema is an important, potentially life threatening complication of lymphoma. Optimal patient management requires that both angioedema and the underlying lymphoma are recognized. PMID- 8242560 TI - Growth patterns of human neuroblastoma xenografts and their relationship to treatment outcome. AB - BACKGROUND: Several investigators have reported the ability to establish xenografts in nude mice from children with neuroblastomas, but a correlation of prognosis with this establishment and the growth patterns of the neuroblastomas has not been reported. METHODS: Tumor specimens from 58 children with neuroblastomas were heterotransplanted into BALB/c nude mice. In 34 patients, heterotransplantation was done before therapy; in 24 patients, tumors were obtained after at least one course of chemotherapy or radiation therapy. The histology, cytogenetics, and growth characteristics of serial passages of the xenografts were studied. RESULTS: The engraftment rate was 34%. Neuroblastomas with diploid chromosome numbers did not engraft. Chromosomal abnormalities involving 1p were seen in more than 50% of the xenografts. Cytogenetic features were retained between original tumors and resultant xenografts. Xenografts could be established only from tumors with unfavorable histology, as defined by Shimada classification criteria. The histology of each xenograft line was strikingly similar, and each was highly undifferentiated. Engraftment rates, doubling times, and lag times did not vary appreciably between xenografts established from treated tumors compared with xenografts established from untreated tumors. There was no correlation between doubling or lag times and prognosis. Patients whose tumors engrafted had only a 5% 3-year survival rate. CONCLUSIONS: From these results, it appears that successful engraftment is the most important prognostic indicator for patients with neuroblastomas. Because of the commonality of the histologic features and the stability of the tumor clones from patients before and after heterotransplantation, these xenografts may be useful as an in vivo model for studying drug resistance and for designing treatment regimens. PMID- 8242561 TI - Malignant paraganglioma presenting as Cushing syndrome with virilism in childhood. Production of cortisol, androgens, and adrenocorticotrophic hormone by the tumor. AB - BACKGROUND: A 12-year-old girl with intractable retroperitoneal paraganglioma experienced increased appetite, acne, obesity, "moon face," and enlargement of the clitoris during the course of the tumor. Plasma cortisol, serum testosterone, and dehydroepiandrosterone sulfate (DHEA-S) levels were increased to 34.1 micrograms/dl, 2.0 ng/ml, and 6.628 ng/ml, respectively. Adrenocorticotrophic hormone (ACTH) levels were not increased, and results of dexamethasone suppression tests were negative. Her condition was diagnosed as Cushing syndrome with virilism. Plasma cortisol levels were increased to a level of 107.1 micrograms/dl before death. METHODS: Tumor samples were obtained at the time of autopsy. The concentrations of cortisol, androgens, ACTH, and catecholamines were assayed in the tumor extracts. The indirect immunoperoxidase procedure was performed on fixed tissues for cortisol, DHEA-S, testosterone, and ACTH. RESULTS: Extracts of the tumor masses contained steroid hormones: the amount of immunoreactive cortisol was 1.64 micrograms/g wet weight; the amount of immunoreactive testosterone was 25.60 ng/g wet weight; immunoreactive DHEA-S, 579.00 ng/g wet weight; and immunoreactive ACTH, 891.00 pg/g wet weight in the metastatic mass of the lung. Immunohistochemically, immunoreactive cortisol, testosterone, and DHEA-S were detectable in the tumor cells. The adrenal gland was atrophic. CONCLUSIONS: The patient is the first reported with malignant paraganglioma with the capacity to produce cortisol, androgens, and ACTH. PMID- 8242562 TI - Human neuroblastoma cell lines that express N-myc without gene amplification. AB - BACKGROUND: About one half of aggressive neuroblastomas lack N-myc amplification. Cell lines from such tumors are needed to determine the biological basis of aggressive tumor behavior. METHODS: Neuroblastoma cell lines were established from a primary tumor (SMS-LHN) and a bone marrow metastasis (LA-N-6) of two children with Stage IV neuroblastoma. Although both cell lines and their original tumors lacked N-myc genomic amplification, these patients died of progressive disease. RESULTS: SMS-LHN and LA-N-6 can be distinguished from primitive neuroectodermal tumor (PNET) lines by cell surface antigen expression and catecholamine production. Cytogenetic analysis of each cell line revealed unique genetic rearrangements, whereas both lines showed abnormalities involving chromosome 2. Neither cell line contained double-minute chromosomes, homogeneously staining regions, a 1p chromosomal deletion, or t(11;22). The growth rates of these two new lines in vitro and in vivo (as xenografts in nude mice) are slower than N-myc amplified neuroblastoma lines. Both lines express greater amounts of N-myc RNA and protein relative to nonneuroblastoma cell lines (including PNET), although not to the extent of cell lines with N-myc genomic amplification. CONCLUSIONS: The relatively large amount of N-myc expression in these two new cell lines suggests that N-myc expression without amplification could play a role in the pathogenesis of some neuroblastomas. These cell lines should be useful for investigating mechanisms and consequences of N-myc gene activation other than genomic amplification. PMID- 8242563 TI - Disclosing the cancer diagnosis. Procedures that influence patient hopefulness. AB - BACKGROUND: Physicians view enhancing patient hopefulness as a critical aspect of cancer treatment, yet little is known about how the circumstances of the initial disclosure of a cancer diagnosis affects this important psychosocial variable. The authors examined the extent to which various forms of physician disclosure of a cancer diagnosis are seen by patients as more or less hopeful and as favorable or unfavorable. METHODS: Statements describing various circumstances of diagnostic disclosure were generated from interviews with 10 physicians and 10 patients with cancer. Fifty-seven statements were rated on two bipolar rating scales describing hopefulness and favorability by an additional 56 patients with recent onset of cancer. To determine if overall emotional adjustment to the illness affected these ratings, scores on the Mental Adjustment Scale were also obtained. RESULTS: Results revealed that patients had clear ideas about which procedures were most and least hopeful and favorable or unfavorable, that the ratings demonstrated clear factorial validity suggesting broad areas of diagnostic disclosure procedure where physicians may make efforts to enhance hope, and that ratings of hopefulness and favorability were related to overall emotional adjustment to the illness. CONCLUSIONS: There was consensus among patients that certain procedures enhanced hopefulness and certain procedures reduced it. Physicians would do well to attend carefully to this aspect of treatment and continually seek patient input concerning their preferences for the conduct of the treatment relationship. PMID- 8242564 TI - Cutaneous malignant melanoma of the head and neck. PMID- 8242565 TI - Spontaneous 19-year regression of oat cell carcinoma with scalene node metastasis. PMID- 8242566 TI - Patterns of presentation of Hodgkin disease. PMID- 8242567 TI - Meta-analysis of radiation therapy with and without adjuvant chemotherapy for malignant gliomas in adults. PMID- 8242568 TI - Primary chemotherapy for obstructive jaundice caused by intermediate-grade non Hodgkin lymphoma. PMID- 8242569 TI - Lung cancer after radiation therapy for breast cancer. PMID- 8242570 TI - Cancer chemoprevention. AB - Encouraging results from several recent trials have increased interest in the potential of chemoprevention in reducing the incidence of cancer. Treatment with 13-cis-retinoic acid has reversed oral leukoplakia, a premalignant lesion, and prevented the development of second primary carcinomas of the head and neck. Tamoxifen has proven effective in preventing both the recurrence of breast cancer and the development of new tumors in the contralateral breast, and is being tested in high-risk patients who have not yet developed disease. The results of the Physicians' Health Study, which demonstrated the effectiveness of aspirin in reducing heart attacks, already have been incorporated into health care practice, while the component investigating the effect of beta carotene on the incidence of cancer still is underway. These studies demonstrate the importance of careful design if chemoprevention trials are to yield convincing data. PMID- 8242571 TI - Hematopoietic cytokines. Current use in cancer therapy. AB - The introduction of hematopoietic cytokines into the clinic has been rapid with three currently approved by the Food and Drug Administration and perhaps a dozen more in clinical trials. Combinations of cytokines have been relatively unexplored in the clinics. Knowledge about the use of cytokines with chemotherapy has grown considerably in the past few years. The literature relating to use of cytokines in support of standard chemotherapy, marrow failure syndromes associated with malignant disease, and dose intensification not requiring progenitor cell replacement has been reviewed. This review does not address the use of cytokines to support bone marrow transplantation and collection of progenitor cells. Hematopoietic cytokines shorten the duration of cytopenia and decrease infectious complications when used to support standard chemotherapy regimens. However, several randomized trials have failed to show a benefit of such cytokine-supported treatment programs in terms of antitumor response, palliation, or survival. One exception is granulocyte-macrophage colony stimulating factor used with aggressive but standard chemotherapy for high-risk non-Hodgkin lymphoma. Cytokines decrease the infectious complications of myelodysplastic syndromes and may decrease transfusion requirements. Marked escalations in chemotherapy doses are possible with specific chemotherapy regimens. These increase complete remission rates although no benefit in survival or palliation has yet been proven. Cytokines have not been effective in allowing escalation of doses with certain chemotherapy agents and regimens. Cytokines decrease hematopoietic toxicity with standard or escalated chemotherapy doses. Randomized trials are now beginning to define the true patient benefit of this capability. PMID- 8242572 TI - The use of colony stimulating factors in marrow transplantation. AB - Colony stimulating factors (CSF) are now widely used in marrow transplantation. Randomized trials have shown that granulocyte macrophage-CSF accelerates marrow recovery after autologous marrow transplantation, resulting in fewer infections, shorter hospitalization, and lower costs. Similar results have been seen with granulocyte-CSF. Both factors also can accelerate engraftment after allogenic marrow transplantation, but there is, so far, less evidence for clinical benefit in this setting. Both granulocyte macrophage-CSF and granulocyte-CSF benefit some patients with graft failure. The recent demonstration that both granulocyte-CSF and granulocyte macrophage-CSF can mobilize large numbers of hematopoietic stem cells into peripheral blood has resulted in the widespread use of this technique as an alternative to autologous marrow transplantation. A number of colony stimulating factors, including IL-1, IL-3, and steel factor, which act on early hematopoietic progenitors, have recently been entered into clinical trials, with the hope that the progress already made with G-CSF and GM-CSF can be continued. PMID- 8242573 TI - Treatment of intractable cancer pain by electronically controlled parenteral infusion of analgesic drugs. AB - Intractable metastatic cancer pain can be controlled in the majority of cases with narcotic drugs. This review focuses on parenteral analgesic therapy; the available drugs, administration systems, and electronic infusion devices. The range of drugs suitable for parenteral therapy, including continuous infusion therapy by subcutaneous, intravenous, or epidural routes is limited both by pharmacokinetic and side effects profiles. Implantable ports and catheter systems permit ambulatory delivery of drugs when combined with portable electronic pumps. This enables patient care to move from a bed-bound hospital therapy to ambulation in the hospital or home. The safeguards, features, and modes of operation of these pump-port-catheter-drug systems are described. PMID- 8242574 TI - Development and implementation of a pain education program. AB - Pain is a significant problem for cancer patients and is of particular concern for the elderly. This article reports on the development and implementation of a pain management educational intervention for elderly cancer patients and their family caregivers. This patient education program was developed within a research study funded by the American Cancer Society. Subjects were stratified as either elderly (60-75 years) or oldest (greater than 75 years) and then were randomly assigned to an experimental group that received the three-part, structured pain education program or a control group that received usual care. Control group subjects were offered the intervention at the conclusion of data collection. The outcomes of the pain education were measured at two times after the intervention. Study variables included patient outcomes such as pain intensity, pain knowledge and attitudes, medication compliance, and quality of life as well as family caregiver outcomes. This pain education program was developed, implemented, and evaluated in 40 patients and family caregivers in the first year of a 2-year project. In this article, the investigators present the development and structure of the three-part education program, general principles of patient education regarding pain, methods of evaluating pain education, and initial results of the pain education. Pain education includes basic principles of pain relief, pharmacologic interventions, and nondrug interventions for relief of pain. This program has demonstrated that both cancer patients and their family caregivers benefit from a structured education program for relief of cancer pain. PMID- 8242575 TI - Strontium-89--precursor targeted therapy for pain relief of blastic metastatic disease. AB - Strontium-89 is a radioactive calcium analog that provides an energetic beta particle for radiation therapy of osteoblastic disease. Strontium-89 is used as palliative therapy with the primary goal being pain relief. More than 500 patients with painful blastic metastatic disease were treated at University of Kansas Medical Center since the initiation of the first clinical trial there 15 years ago. Most patients have had metastatic prostate cancer to bone or breast cancer, as these tumors are commonly associated with bone pain as their primary clinical management problem. Improvement (decrease in pain, increase in physical activity level) was noted in 80% of patients with prostate carcinoma and 81% of patients with metastatic breast cancer to bone. Marrow toxicity levels were acceptable. The therapy can be repeated at 3-month intervals. Strontium-89 is a safe and effective systemic therapy for painful blastic metastatic disease. There is no longer any reason why the vast majority of persons with painful blastic metastatic disease should continue to hurt. PMID- 8242576 TI - Enhancing the effectiveness of the specific serotonin antagonists. Combination antiemetic therapy with dexamethasone. AB - Combinations of drugs have become standard therapy for the prevention of vomiting caused by anticancer drugs like cisplatin. Recently, a new class of antiemetic agents, the potent and specific 5-HT3 receptor antagonists such as ondansetron, granisetron, and tropisetron, have been shown to be more effective and better tolerated than metoclopramide. This report describes the rationale for combination antiemetic therapy, details the testing of metoclopramide-based regimens as a model for combination therapy development, reviews completed trials of ondansetron plus dexamethasone, and offers strategies to further alleviate vomiting during anticancer chemotherapy. The reported trials testing metoclopramide-based combinations were reviewed and that experience was applied to the ongoing studies of ondansetron when used with dexamethasone and other agents. Combinations of metoclopramide, dexamethasone, and lorazepam prevented acute emesis caused by high-dose cisplatin in 63% of patients, lessened side effects, and were convenient enough to administer to outpatients. Completed trials of ondansetron and dexamethasone demonstrated improved vomiting control over ondansetron alone while using less cumbersome schedules. Attempts to improve ondansetron-based antiemetic regimens by developing optimal drug doses and schedules and adding adjuvant and different classes of antiemetic agents are now in clinical testing. Based on previous experience and current results, combinations of a specific serotonin agonist and dexamethasone are the best treatment for prevention of vomiting induced by chemotherapy. Future clinical research should aim to refine antiemetic regimens and improve emetic control through the use of new antiemetic and adjuvant agents. PMID- 8242577 TI - New bisphosphonates in the treatment of bone metastases. AB - Normal skeletal integrity is maintained by physiological bone turnover through a coupled process of bone resorption, mediated by osteoclasts, followed by new bone formation, mediated by osteoblasts. Major features of the pathogenesis of cancer associated skeletal destruction are enhanced osteoclast-mediated bone resorption and disruption of normal bone formation. In this article, the literature on the pathogenesis and clinical manifestations of metastatic bone disease is discussed. Animal and clinical trials investigating novel bone targeted agents, emphasizing the bisphosphonates, are critically assessed. The most frequent clinical manifestations of bone metastases are pain, fracture, immobility, spinal cord compression, and hypercalcemia. New treatments under study for patients with bone metastases include agents specifically targeted to the skeleton such as bone seeking radioisotopes and bisphosphonates. Studies in animal models of metastatic bone disease show that these bisphosphonates are able to inhibit tumor-induced osteolysis and are potentially useful in this condition. Bisphosphonates have been investigated in several clinical trials of patients with skeletal metastases from breast cancer, prostate cancer, and multiple myeloma. Overall, the studies investigating bone targeted radioisotopes or bisphosphonates for the treatment of morbidity due to skeletal metastases have been inconclusive. An improved understanding of the pathogenesis of metastatic bone disease and preclinical studies with bisphosphonates suggest that these agents may have a role in the treatment of this disorder. Additional trials of new generation bisphosphonates, employing a rigorously controlled, randomized study design with adequate numbers of subjects, are needed to demonstrate the safety and efficacy of this class of agents in this setting. PMID- 8242578 TI - Immunoscintigraphy in patients with colorectal, ovarian, and prostate cancer. Results with site-specific immunoconjugates. AB - Application of monoclonal antibody (MoAb) technology to cancer management is discussed by reviewing the development and clinical evaluation of two MoAb-based immunoscintigraphic agents (111In-satumomab pendetide [OncoScint CR/OV-In] and 111In-CYT-356; Cytogen Corporation, Princeton, NJ). Both agents were prepared using a site-specific MoAb modification method that preserves the immunoreactivity of the radiolabeled immunoconjugate. 111In-satumomab pendetide is an 111In-labeled conjugate of the murine MoAb B72.3, which is directed to TAG 72, an antigen expressed by the majority of adenocarcinomas. By providing information that complements the results of standard radiographic diagnostic modalities, this imaging agent can aid in the treatment of patients with colorectal or ovarian cancer. Immunoscintigraphy with 111In-satumomab pendetide has been shown to assist in medical-surgical management by detecting occult extrahepatic lesions, clarifying equivocal results of other diagnostic imaging tests, and evaluating the extent and resectability of known tumor lesions. 111In CYT-356 is an 111In-labeled conjugate of the murine MoAb 7E11-C5.3, which is reactive with prostatic carcinoma, benign prostatic hypertrophy, and, to a lesser extent, normal prostatic tissue. Results of preliminary clinical investigations suggest that 111In-CYT-356 immunoscintigraphy can be useful for the presurgical staging of prostatic carcinoma and for the detection of occult distant disease in patients with negative or equivocal results on standard imaging tests. Results with these site-specifically radiolabeled immunconjugates demonstrate the clinical utility of MoAb-based imaging agents in the treatment of patients with solid tumors. PMID- 8242579 TI - Therapeutic advances in radiologic treatment of cancer. AB - In 1993, roughly 1,170,000 new cases of invasive cancer will be diagnosed in the United States. A considerable portion of these patients who die will do so because of uncontrolled local and/or regional disease. Numerous advances in the radiologic treatment of cancer have led to improved local and regional control, as well as survival. These advances include improved radiologic imaging techniques, improved techniques of radiation therapy, innovative brachytherapeutic techniques, intraoperative electron beam therapy techniques, and stereotaxic radiosurgical techniques. In this paper, the authors discuss these techniques with specific attention to how they have lead to improved local and regional control, as well as cancer cure with organ preservation. PMID- 8242580 TI - Reversal of multidrug resistance to cancer chemotherapy. AB - In the past few years, the role of membrane-bound transport genes in human disease has been increasingly recognized and understood. Of these genes, the p170 membrane glycoprotein may function as an outward transport pump for many cancer chemotherapeutic drugs associated with the multidrug resistance phenotype. This article reviews the different classes of the current major modulators of multidrug resistance. PMID- 8242581 TI - Newer approaches to the management of multiple myeloma. AB - Patients with multiple myeloma must be distinguished from those with monoclonal gammopathy of undetermined significance and smoldering multiple myeloma. Therapy should be reserved for patients with active or symptomatic multiple myeloma. The pertinent literature on the diagnosis of multiple myeloma, prognostic factors, chemotherapy, and allogeneic bone marrow transplantation, as well as autologous peripheral blood or bone marrow stem cells for rescue, was reviewed. The two most powerful prognostic factors for multiple myeloma are the bone marrow plasma cell labeling index and the beta 2-microglobulin level. Chemotherapy is the preferred initial treatment for overt, symptomatic multiple myeloma. Combinations of alkylating agents produce a higher response rate, but the survival is the same as treatment with melphalan and prednisone. The combination of alpha 2-interferon with multiple alkylating agents produces a good response. alpha 2-Interferon prolongs the duration of the plateau state after a response to chemotherapy, but it apparently does not prolong survival. Allogeneic bone marrow transplantation is possible for only 5-10% of patients with multiple myeloma. Its advantage is that the graft contains no tumor cells that can subsequently produce a relapse. However, there is a significant early mortality, the risk of graft versus host disease is troublesome, and relapse of multiple myeloma is common. Autologous bone marrow transplantation is applicable for more patients because the age limit is higher and a matched donor is unnecessary. However, two major problems exist: (1) eradication of multiple myeloma from the patient may not occur even with large doses of chemotherapy and irradiation, and (2) infused autologous bone marrow or peripheral blood stem cells contaminated by myeloma cells or their precursors may be responsible for relapse. PMID- 8242582 TI - Amifostine-mediated protection of normal bone marrow from cytotoxic chemotherapy. AB - Amifostine (US Bioscience, West Conshohocken, PA; Ethyol, WR-2721), a phosphorylated thiol developed by the United States Army as a protective agent for military personnel in the event of nuclear warfare, has shown protection of normal tissues from the cytotoxic effects of therapeutic radiation and chemotherapy with preservation of cytotoxic effects on the tumor. The basis of this selective protection derives from the relatively rapid uptake and anabolism of Amifostine into normal tissues and minimal, slower uptake into tumor tissue. Preclinical investigations have demonstrated protection of bone marrow stem cells from the toxic effects of radiation and chemotherapy. Several controlled clinical trials demonstrated this hematoprotective effect. In patients given 1.5 g/m2 cyclophosphamide and month later given Amifostine (740 mg/m2) followed by the same dose of cyclophosphamide, the median nadir neutrophil count was significantly increased and duration of neutropenia was significantly reduced by pretreatment with Amifostine. In women with stage III/IV ovarian cancer treated with 1 g/m2 cyclophosphamide and 100 mg/m2 cisplatin +/- Amifostine 910 mg/m2, treatment with Amifostine before cyclophosphamide and cisplatin resulted in a significant decrease in both the incidence and duration of hospital stays for neutropenic fever compared to cyclophosphamide and cisplatin alone. There were equivalent rates of response and duration of survival in both groups. Other studies have shown Amifostine protects bone marrow purged in vitro with 4 hydroperoxycyclophosphamide before autologous bone marrow transplantation. This preservation of marrow stem cells resulted in a statistically significant decrease in time to marrow engraftment, need for platelet transfusions and antibiotics, and duration of hospital stay. Amifostine-mediated protection of normal bone marrow illustrated in preclinical experiments is also evident in clinical trials. Amifostine preserves trilineage stem cells (red blood cells, platelets, and white blood cells) in contrast to the lineage-specific effects of the colony-stimulating factors. Theoretically, Amifostine and the colony stimulating factors should provide complementary benefits to bone marrow recovery and function after cytotoxic therapies. These observations offer the promise of using high doses of chemotherapy to exploit antitumor, dose-response relationships in clinical trials. PMID- 8242583 TI - Neoadjuvant therapy in cancer treatment. AB - Neoadjuvant therapy has come to play an increasingly prominent role in the treatment of cancer. Originally defined as systemic therapy given before local treatment, the concept has been extended to include radiation therapy given before surgery. Potential advantages include improved local and distant control, direct evaluation, and organ-sparing treatment. Potential disadvantages include increased toxicity and cost, potential delay in effective treatment, and obscuring of pathologic staging. Neoadjuvant therapy in cancer treatment may be viewed in three categories: tumors in which neoadjuvant treatment has been shown effective, thus becoming standard therapy; tumors in which it has been shown to facilitate organ-sparing, and tumors in which its utility has not been shown. For patients with osteogenic sarcoma, for example, preoperative chemotherapy and limb salvage therapy have become the standard of care. Response to chemotherapy, ascertained by histologic review of the surgical specimen, can be used to tailor postoperative chemotherapy. In patients with advanced laryngeal squamous cell carcinoma, neoadjuvant chemotherapy followed by radiation has permitted laryngeal preservation in a majority of patients without compromising overall survival. Phase II and III studies conducted in women with breast cancer have demonstrated promising results for neoadjuvant chemotherapy given before radiation therapy and/or surgery. Phase III studies to compare neoadjuvant therapy to standard therapy in patients with breast cancer are underway. For neoadjuvant therapy, as with other innovations in cancer treatment, it is crucial that a new strategy must be compared closely to standard therapy in terms of recurrence, survival, and impact on organ sparing, as well as quality of life and treatment costs. PMID- 8242584 TI - A model system. Integration of services for cancer treatment. AB - The care of pediatric cancer patients continues to grow in complexity. Paradoxically, treatment regimens grow more intensive, while regulatory pressures mandate more outpatient care. The challenge is to integrate services around episodes of illness and encounters over periods of 1-3 years. The approach of the author's clinic has been to create seamless relationships that place the patient at the center of care and address the major boundaries patients face. The inpatient-outpatient boundary has been effectively breached by an inpatient case manager and the simultaneous temporary rotation of an inpatient nurse to the outpatient area for specialty training. Discharge planning has been improved by sharing a nurse with the clinic's major home health care company, providing a direct clinic-home health liaison for patients. Ongoing formal evaluations have documented the effects of the institution of each part of the program. These surveys have indicated that inpatient staff participating in the outpatient rotation are more satisfied with the continuity of care and the availability of divisional resources. Patient satisfaction was extremely high and pediatric oncology discharge planning was rated significantly higher than other pediatric services. Despite rapid growth of the oncology service since 1985 and the personal intensity of care, length of stay has shortened and the number of staff has not required excessive increases to meet the needs of the new model. PMID- 8242585 TI - Home parenteral and enteral nutrition in cancer patients. AB - The clinical outcomes of 2968 patients with active cancer receiving home nutrition support are described. Of these patients 1672 were receiving home parenteral nutrition and 1296 were receiving home enteral nutrition. The outcomes of these active cancer patients are compared to those of 123 radiation enteritis ("cured" cancer) and 480 Crohn's disease patients receiving home parenteral nutrition and 918 noncancer dysphagic patients receiving home enteral nutrition. This longitudinal clinical information was reported to the North American Home Parenteral and Enteral Nutrition Patient Registry between 1985 and 1990. Evidence indicates that the number of home parenteral and enteral nutrition patients has increased nationally by about 25% each year between 1989 and 1990. In a subsample of 37 home nutrition support programs that have consistently reported their data to the registry since 1985, more than 90% of their program growth was accounted for by new patients with active cancer. This is now the largest single diagnosis of patients starting home parenteral and enteral nutrition. The mean survival time of cancer patients is 6 months after starting home parenteral and enteral nutrition, but 25% live beyond a year and 20% resume full oral nutrition. Although most active cancer patients experience only partial rehabilitation, for those patients with longer survival, rehabilitation is more complete. The outcome is relatively better for children and for patients whose neoplasm is leukemia, lymphoma, small bowel, or liver. In comparison, 96% of home parenteral nutrition Crohn's patients, 80% of home parenteral nutrition radiation enteritis patients, and 60% of home enteral nutrition noncancer dysphagic patients survive at least 1 year. Adult active cancer home parenteral and enteral nutrition patients do not have a greater incidence of therapy-related readmissions than other patient groups, however, their overall rehospitalization rate is much more frequent. This article discusses factors that may have contributed to this growth in home nutrition support in active cancer patients. It attempts to clarify where this therapy is clearly justified and where more information is needed. It emphasizes some of the special issues that need to be addressed in treating these vulnerable patients. PMID- 8242586 TI - A clinician-driven home care delivery system. AB - The financial, entrepreneurial, administrative, and legal forces acting within the home care arena make it difficult for clinicians to develop and operate home care initiatives within an academic setting. HomeMed is a clinician-initiated and -directed home care delivery system wholly owned by the University of Michigan. The advantages of a clinician-directed system include: Assurance that clinical and patient-based factors are the primary determinants of strategic and procedural decisions; Responsiveness of the system to clinician needs; Maintenance of an important role for the referring physician in home care; Economical clinical research by facilitation of protocol therapy in ambulatory and home settings; Reduction of lengths of hospital stays through clinician initiatives; Incorporation of outcome analysis and other research programs into the mission of the system; Clinician commitment to success of the system; and Clinician input on revenue use. Potential disadvantages of a clinician-based system include: Entrepreneurial, financial, and legal naivete; Disconnection from institutional administrative and data management resources; and Inadequate clinician interest and commitment. The University of Michigan HomeMed experience demonstrates a model of clinician-initiated and -directed home care delivery that has been innovative, profitable, and clinically excellent, has engendered broad physician, nurse, pharmacist, and social worker enthusiasm, and has supported individual investigator clinical protocols as well as broad outcomes research initiatives. It is concluded that a clinician-initiated and -directed home care program is feasible and effective, and in some settings may be optimal. PMID- 8242587 TI - Identification of masked and variant Ph (complex type) translocations in CML and classic Ph in AML and ALL by fluorescence in situ hybridization with the use of bcr/abl cosmid probes. AB - In the present study, three chronic myelogenous leukemia (CML) patients with variant Philadelphia (Ph) chromosomes (complex types), two CML patients with a masked Ph, one case with Ph positive acute lymphocytic leukemia (ALL), and one with Ph positive acute myelocytic leukemia (AML) were analyzed by standard cytogenetic techniques (G-banding), Southern blot studies, and fluorescence in situ hybridization (FISH) procedures using probes from portions of the bcr and abl genes. It has been previously shown that this FISH approach could detect the bcr/abl fusion event in CML patients with classic or variant (simple type) Ph. Our results demonstrate that this FISH assay can also detect the bcr/abl fusion status in CML patients with masked or variant (complex type) Ph chromosomes and in some patients with Ph positive ALL or AML. PMID- 8242588 TI - Cosmid mapping and locus linkage within the human chromosomal region 22q13.1. AB - Many human meningiomas show loss of heterozygosity at distal loci but retain constitutional heterozygosity at one or more proximal loci of 22q. Molecular analysis indicted deletions involving at least the region 22q12.3-qter. In this region, distal to myoglobin, the putative meningioma locus ought to be expected. Long-range mapping was performed around two loci from 22q12.3-q13.1 (D22S16 and PDGFB, the most proximal locus to be lost in meningioma). D22S16, originally assigned to 22q13-qter by isotopic in situ hybridization, was placed in the vicinity of PDGFB by utilizing a set of somatic cell hybrids, an assignment confirmed by fluorescence in situ hybridization (FISH) of a cosmid clone containing the D22S16 locus. Moreover, pulsed field gel electrophoresis suggests a close linkage of both markers within 630 kb. PMID- 8242589 TI - Hypothesis: meiotic origin of trisomic neoplasms. AB - The gain of single additional chromosomes is a very common finding among the nonrandom abnormalities with human neoplasms. According to the current opinion, such trisomies result from a disease-related mitotic nondisjunction. In contrast, we suggest that some of these trisomies may in fact represent tissue-confined residual cell populations of meiotic origin. Our hypothesis is based on recent findings of uniparental disomies in humans (i.e., two homologous chromosomes are derived from the same parent) and on the notion that many of these disomies probably are the result of the successful correction of an initially trisomic conceptus. Thus, the trisomic neoplasm may represent the original trisomic karyotype, whereas the apparently normal disomic constitutional karyotype may be the acquired, corrected one. We propose molecular genetic strategies to test our hypothesis and suggest that constitutional uniparental disomies may be associated with an increased probability of developing neoplastic diseases characterized by trisomies for the respective chromosomes. PMID- 8242590 TI - Therapy-related myelodysplastic syndrome. Two cytogenetically unrelated abnormal clones in a patient with multiple myeloma. AB - A multiple myeloma patient presented for cytogenetic analysis at diagnosis of secondary MDS, which followed cytotoxic treatment including melphalan. Two abnormal unrelated clones were detected, one of them had 5q-, 7q- with clonal evolution of an additional aberration, t(12;13); in the second clone there was a translocation between the two homologues of chromosome 1 as the only aberration. We suggest that the clone with 5q- and 7q- represented the secondary MDS cells, whereas the abnormal clone with t(1;1) represented the plasmablasts of the multiple myeloma. PMID- 8242591 TI - Cell lineage involvement in four patients with myelodysplastic syndrome and t(1;7) or trisomy 8 studied by simultaneous immunophenotyping and fluorescence in situ hybridization. AB - Four patients with myelodysplastic syndrome (MDS), one with t(1;7) and three with trisomy 8, were studied by immunophenotyping and fluorescence in situ hybridization (FISH) to assess cell lineage involvement. The t(1;7) was detected using a biotin-labeled chromosome 1 centromere-specific DNA probe. This aberration was present in CD34-positive stem cells, the erythroid cell lineage (GPA+), and the granulocytic/monocytic (CD13+ and CD64+) cell lineages. We were not able to demonstrate the abnormality in the lymphoid cell lineages. In the patients with trisomy 8, the aberration was detected with chromosome 8 centromere specific DNA probe or by chromosome in situ suppression hybridization (CISS) with a chromosome 8-specific library probe. The trisomy was detected in stem cells, erythroid precursor cells, megakaryocytes, and granulocytes/monocytes. In these MDS patients, the chromosome aberrations appear to occur only in cells of myeloid lineage. PMID- 8242592 TI - t(8;21) prior to acute leukemia. AB - A t(8;21)(q22;q22) without blood and bone marrow invasion by immature myeloid precursor cells occurred in a patient previously treated for polycythemia vera. The presence of a molecular rearrangement confirmed that the chromosomal abnormality was identical to that observed in acute leukemia with t(8;21). This case shows that the translocation, t(8;21), may occur in myelodysplasia and suggests that it can precede the appearance of overt leukemia. PMID- 8242593 TI - Cytogenetic studies of primary cultures of esophageal squamous cell carcinoma. AB - Cytogenetic analysis was performed on primary cultures of 21 squamous cell carcinomas of the esophagus (SCCE). Seven cases exhibited mosaic clonal chromosome abnormalities distributed as follows: two contained tetraploid cell populations, one with t(3;7)(p21;q11); two showed loss of the Y chromosome, one with double minutes; single cases demonstrated der(11)t(4;11)(q?27;q23); add(1)(p35) and del(4)(p12); and del(7)(p13), del(7)(q22q34), and der(11)t(7;11)(p?15;p?13). The remaining 14 cases had apparently normal karyotypes, possibly derived from stromal elements. These results demonstrate numerical abnormalities and the multiple occurrence of rearrangements involving chromosomes 7 and 11 in SCCE. PMID- 8242594 TI - Presence of telomeric sequences on deleted chromosomes and their absence on double minutes in cell line HL-60. AB - The issue of telomeric sequences on deleted chromosomes and double minutes (dmin) was investigated by examining the cell line HL-60 with fluorescence in situ hybridization using a human plasmid DNA sequence with 800 bp TTAGGG repeats. This cell line showed telomeric sequences on the deleted short arms of chromosomes 9 and 10, with the results suggesting that so-called terminal deletion may be, in fact, an interstitial deletion, or that telomeric sequences may be synthesized by telomerase after deletion. On the other hand, numerous dmin showed no evidence of hybridization with the telomeric probe. This suggests that the characteristics unequal distribution of dmin during mitosis may result from the lack of not only centromeres but also telomeres. PMID- 8242595 TI - Translocation (Y;1)(q12;q21) in acute leukemia. AB - This report documents one patient with myelodysplasia evolving into acute leukemia who showed a t(Y;1) translocation confirmed by in situ hybridization. Most of the q arm of the Y chromosome was translocated to an additional q arm of chromosome 1, resulting in trisomy 1q. To our knowledge only four other cases with this t(Y;1) have been reported. PMID- 8242596 TI - Translocation (2;4)(p23;q25): an additional case of a new recurrent anomaly in acute myeloid leukemia. AB - A second case of acute myeloid leukemia (AML) with a t(2;4)(p23;q25) as the sole anomaly is reported. Our case had a de novo AML (M2); the case previously described had AML (M2) post-MDS. It is suggested that t(2;4)(p23;q25) is a new, recurrent, but rare anomaly in AML. PMID- 8242597 TI - Acute non-lymphocytic leukemia with t(16;21). AB - A patient with ANLL FAB subtype M1 was found to possess a t(16;21)(p11;q22) and trisomy 10. The 16;21 translocation has been reported in 12 other cases of ANLL, of various subtypes, and its relationship to the disease profile is discussed. PMID- 8242598 TI - 5q- anomaly preceding acute myeloblastic leukemia with 53 chromosomes without chromosome 5 abnormality. AB - Partial deletion of the long arm of chromosome 5 (5q-) was found in a patient with a myeloid disorder. After treatment, hematologic parameters became normal, but acute myeloblastic leukemia occurred after 58 months. At that time, cytogenetic studies showed the presence of an abnormal clone with 53 chromosomes and without 5q-. This exceptional case suggests that the 5q- abnormality may precede the occurrence of an acute leukemia in favoring the appearance of unrelated malignant clones. PMID- 8242599 TI - Two Down syndrome patients with an acquired translocation, t(8;14)(q11;q32), in early B-lineage acute lymphoblastic leukemia. AB - Two males with Down syndrome and acute lymphoblastic leukemia with the acquired translocation, t(8;14)(q11;q32), are described. In each case the constitutional karyotype was 47,XY,+21. The patients were, respectively, aged 3 years 11 months and 32 years, with presenting white blood counts 34 and 1.9 x 10(9)/L with blasts of FAB L1 and L2. In each case immunophenotype of the blasts was C-ALL. The child is alive and well and in first remission 6 years from diagnosis. In contrast, the adult patient died in first remission 8.5 months from diagnosis with severe pancytopenia. These are to our knowledge the second and third cases of ALL with t(8;14)(q11-12;q32) associated with a constitutional genetic disorder. PMID- 8242600 TI - Trisomy 4 in a patient with acute myelomonocytic leukemia (M4) PMID- 8242601 TI - Study of the BCR/ABL rearrangement in patients with two Philadelphia chromosomes. PMID- 8242602 TI - der(Y)t(Y;1) is a nonrandom abnormality in myelodysplastic syndrome. PMID- 8242603 TI - Overrepresentation of chromosome 12p sequences and karyotypic evolution in i(12p) negative testicular germ-cell tumors revealed by fluorescence in situ hybridization. AB - Human testicular germ-cell tumors (TGCTs) comprise a heterogeneous group of solid neoplasms. These tumors are characterized by the presence of a highly specific chromosomal abnormality, i.e., an isochromosome of the short arm of chromosome 12. At present, this i(12p) chromosome is found in more than 80% of TGCTs. Isochromosome 12p has also been observed in some ovarian and extragonadal germ cell tumors. In the remaining so-called i(12p)-negative TGCTs other abnormalities involving chromosome 12, mainly 12p, can be found. In order to establish whether 12p abnormalities other than i(12p) are a common phenomenon in TGCTs, a panel of 11 i(12p)-negative tumors was investigated using multicolor fluorescence in situ hybridization. All TGCTs examined appeared to contain chromosomal abnormalities involving 12p, resulting in a distinct overrepresentation of short arm sequences. In addition, indications were obtained for a clonal evolution in one of the tumors. Our data suggest that the occurrence of 12p abnormalities is a common phenomenon in i(12p)-negative TGCTs and that these abnormalities, analogous to i(12p), may contribute to the process of tumor development. PMID- 8242604 TI - Characterization of two marker chromosomes in a patient with acute nonlymphocytic leukemia by two-color fluorescence in situ hybridization. AB - A patient with acute nonlymphocytic leukemia (ANLL), M5b according to French American-British (FAB) classification, showed monosomy 16, an extra 1p-, and a 21q+. These derivative chromosomes could not be defined by GTG-banding. For better characterization, we performed two-color fluorescence in situ hybridization (FISH) experiments applying DNA libraries from sorted human chromosomes, chromosome-specific repetitive probes, and a band-specific YAC clone. With these FISH studies the karyotype could be characterized as 46,XY, +der(1)t(1;21)(p11;?), -16,der(21)t(16;21) (p11.1;q22). PMID- 8242605 TI - A rapid screening method to detect nonsense and frameshift mutations: identification of disease-causing APC alleles. AB - A functional screen for nonsense and frameshift mutations has been devised that allows genes of interest to be scanned in segments. This assay is based on the cloning of these segments in-frame with a colorimetric marker gene (lacZ) followed by screening for the level of functional activity from the marker polypeptide (beta-galactosidase). Individuals at risk for any one of a number of genetic diseases, in particular familial adenomatous polyposis coli (APC), can be quickly screened for chain-terminating mutations introduced by stops and frameshifts. At present, scanning of the APC gene for mutation requires significant effort because it is a large gene and most APC mutations are unique. Therefore, this assay offers a powerful option for the diagnosis of this and other genetic diseases, as well as great potential for the development of a similar rapid screen to detect APC mutations in colorectal adenomas and carcinomas. PMID- 8242606 TI - Insulin-like growth factor I gene expression in the uterus is stimulated by tamoxifen and inhibited by the pure antiestrogen ICI 182780. AB - Estrogen-induced uterine insulin-like growth factor I (IGF-I) expression has been demonstrated to mediate at least in part the uterotrophic action of estradiol. We studied the effects of tamoxifen, a partial antagonist to the estrogen receptor widely used in the treatment of breast cancer, and ICI 182780, a pure antagonist to the estrogen receptor, on uterine weight and uterine IGF-I gene expression in the rat. Tamoxifen increased uterine weight to 125% of control values and doubled uterine IGF-I expression. In contrast, ICI 182780 reduced uterine weight to 60% of control and uterine IGF-I gene expression to 13% of control. These results demonstrate for the first time that uterine IGF-I expression is a molecular marker that correlates with the effects of partial agonists and antagonists to the estrogen receptor on the uterus. Furthermore, the induction of uterine IGF-I expression by tamoxifen provides a molecular mechanism to account for the uterotrophic effects which are commonly seen with tamoxifen therapy and which have been associated with endometrial neoplasia. PMID- 8242607 TI - APC gene messenger RNA: novel isoforms that lack exon 7. AB - The APC gene at human chromosome 5q21 is responsible for familial adenomatous polyposis coli. Furthermore, sporadic cancers of not only colon but also other digestive organs often contain mutations in the APC gene. A dominant mouse mutation Min that was generated by chemical mutagenesis and causes polyposis in the digestive tract is in the mouse homologue of the human APC gene. The APC mRNA is generated from 15 exons. Two mRNA isoforms were reported which are produced by alternative splicing in the 9th exon. Here, we report novel mRNA isoforms that lack the 7th exon in both mouse and human cells. PMID- 8242608 TI - Mitosis-promoting factor activity of inducer mitotic cells may affect radiation yield of interphase chromosome breaks in the premature chromosome condensation assay. AB - We measured mitosis-promoting factor (MPF) activity in two cell lines, CHO and HeLa, extensively used at mitosis as inducers in the assay of premature chromosome condensation to study the yield and the repair kinetics of radiation damage in interphase chromosomes of diverse cell lines. We found a 2.5-fold higher MPF activity in HeLa as compared to CHO mitotic cells per mg of crude extract protein. HeLa mitotic cells, when used as inducers of premature chromosome condensation, uncovered two times more interphase chromosome breaks in irradiated, nonstimulated human lymphocytes as compared to CHO mitotic cells. A 2 fold increase in the yield of interphase chromosome breaks with HeLa mitotics was also observed in G1 cells from plateau-phase CHO cultures. Thus, MPF activity may be a contributing factor of the process that transforms radiation-induced DNA damage to chromosome breaks, and subsequently to other types of lethal chromosome aberrations. We speculate that the level and the control in the cell cycle of MPF activity may influence the radiosensitivity of cells to killing. The results strongly suggest that a direct comparison between the yields of interphase chromosome breaks measured in different laboratories may not be possible unless similar inducer cells with similar MPF activity are used. PMID- 8242609 TI - Prostatic nerve growth factor inducible A gene binds a novel element in the retinoblastoma gene promoter. AB - We are investigating the role of the early response transcription factor, nerve growth factor inducible A gene (NGFI-A), as a modulator of retinoblastoma (RB) gene transcription in prostate cells. Examination of the RB promoter reveals a novel element GCGGGGGAG located at nucleotides 152-144 upstream of the methionine initiation codon. This sequence shares strong homology with the consensus NGFI-A binding element GCGGGGGCG varying by a single nucleotide. In DNA binding assays, an NGFI-A fusion protein and the native protein product of the NGFI-A gene purified from prostate cancer cells bound specifically to an oligonucleotide containing the RB promoter element. Gene expression studies in rat ventral prostate demonstrated a 1.9-fold increase in RB mRNA following castration that parallels a 2.7-fold induction of NGFI-A mRNA. In summary, the in vitro DNA binding data and the transient coregulation of rat NGFI-A and RB following castration suggests that the RB gene may be transcriptionally regulated by NGFI-A in prostate cells. PMID- 8242610 TI - Deacetylase activity of human tumor cells producing immunosuppressive aminosugars: its possible role in resistance to cell-mediated cytotoxicity. AB - In the present study, we examined the presence of deacetylases capable of producing free hexosamines, which we have shown earlier to be immunosuppressive against human natural killer (NK) cell-mediated cytotoxicity, from N acetylhexosamines in human tumor cells. When human NK-resistant colon cancer cells (Colo-320DM) were incubated with acetyl-D-[1,6-3H(N)]glucosamine, a significant conversion to [3H]glucosamine occurred. Deacetylation was demonstrated as a change of the substrate radioactivity into free glucosamine trapped by a cation exchange resin, and this was subsequently confirmed by paper chromatography. This deacetylase activity was detected in other NK-resistant tumor cell lines, especially in freshly isolated human renal and breast cancer cells and testicular seminoma cells. However, no deacetylase activity was detected in NK-sensitive target cells such as K562, MOLT-4, or HL-60 cells. The ability to produce free hexosamines from N-acetylated aminosugars may provide a new mechanism for the escape of tumor cells from the attack of immune effector cells such as NK cells. PMID- 8242611 TI - Expression of the tumor suppressor gene DCC in human gliomas. AB - Reduced expression and/or allelic loss of the putative tumor suppressor gene DCC has been demonstrated in colorectal, gastric, pancreatic, esophageal, breast, and hematological malignancies. We examined the expression of the DCC gene in 22 tissue samples from human gliomas (glioblastoma multiforme, oligodendroglioma, and mixed oligodendroglioma/astrocytoma). Seven of 8 glioblastomas multiforme (88%) had reduced or absent DCC expression, and 8 of the other 14 tumors underexpressed DCC when compared to normal brain tissue. These results demonstrate that reduced expression of DCC occurs in human malignant gliomas and may be part of a common genetic pathway leading to neoplastic transformation and/or tumor progression. PMID- 8242612 TI - Decreased expression of the signal-transducing zeta chains in tumor-infiltrating T-cells and NK cells of patients with colorectal carcinoma. AB - An impaired immune response is frequently observed in cancer patients and tumor bearing mice. T-cells from mice with an experimental colon carcinoma were recently shown to express T-cell receptors that completely lacked the signal transducing molecule CD3 zeta. Here, we have investigated the expression of the signal-transducing molecule zeta on lymphocytes from 14 patients with colorectal carcinomas using flow cytometric analysis of permeabilized cells with a monoclonal antibody (TIA-2; IgG1) specific for the cytoplasmic domain of the zeta chain as well as with immunoprecipitation and analysis on diagonal gel electrophoresis. We demonstrate that T-cells isolated from the tumors of the patients express significantly less CD3 zeta than T-cells in the peripheral blood of the same patients and that the peripheral blood of the patients express decreased levels of zeta chains, as compared to the levels found in lymphocytes from healthy controls. This decreased expression was also observed on zeta chains associated with the low affinity Fc receptor for IgG found in tumor-infiltrating NK cells (Fc gamma RIIIA alpha; CD16). PMID- 8242613 TI - Loss of T-cell receptor zeta chain and p56lck in T-cells infiltrating human renal cell carcinoma. AB - Cancer patients and mice bearing tumors develop a progressive immunosuppression manifested by a decreased delayed-type hypersensitivity, decreased T-cell lytic activity, diminished production of lymphokines, and a reduced T-cell proliferative response. The mechanisms underlying these changes are incompletely understood. We recently reported the presence of marked alterations in signal transduction in T-cells from mice bearing long-term (28-day) tumours. We hypothesized that a soluble product produced by the tumor or resulting from the immune response to tumor might be responsible for inducing the changes in T cells. Tumor-infiltrating lymphocytes from patients with renal cell carcinoma tested here showed, in 10 of 11 cases, a marked decrease in the expression of the T-cell receptor zeta chain and in p56lck tyrosine kinase. The presence of major alterations in the tumor-infiltrating lymphocytes with only minor changes in the peripheral blood leukocyte T-cells supports the notion that the defects are induced by exposure to tumor. These results suggest that tumor-infiltrating lymphocytes may be compromised in their antitumor efficacy in patients with renal cell cancer. PMID- 8242614 TI - Identification of three regions on chromosome 17q in primary human breast carcinomas which are frequently deleted. AB - We have examined the long arm of chromosome 17 in sporadic breast carcinomas for the loss of heterozygosity (LOH) at 18 polymorphic loci. At least three distinct regions could be identified by the frequency of LOH and confirmed by high density deletion maps of individual tumor DNAs. A proximal region affected by LOH is located in a 22-cM region defined by D17S73 and NME1 and thus is similar in location to the region thought to contain the BRCA1 gene associated with familial breast and breast/ovarian cancer. The central region affected by LOH is bordered by the D17S86 and D17S21 loci and is estimated to be 28 cM in size. The third region is bordered by the D17S20 and D17S77 loci which are 11 cM apart. These results define three independent regions of chromosome 17q which are likely to contain tumor suppressor genes relevant to the etiology of sporadic breast carcinoma. PMID- 8242615 TI - Microsatellite instability in bladder cancer. AB - Somatic instability at microsatellite repeats was detected in 6 of 200 transitional cell carcinomas of the bladder. Instabilities were apparent as changes in (GT)n repeat lengths on human chromosome 9 for four tumors and as alterations in a (CAG)n repeat in the androgen receptor gene on the X chromosome for three tumors. Single locus alterations were detected in three tumors, while three other tumors revealed changes in two or more loci. In one tumor we found microsatellite instability in all five loci analyzed on chromosome 9. The alterations detected were either minor 2-base pair changes or larger (> 2 base pairs) alterations in repeat length. All six tumors were low stage (Ta-T1), suggesting that these alterations can occur early in bladder tumorigenesis. PMID- 8242616 TI - Cloning of the ALL-1 fusion partner, the AF-6 gene, involved in acute myeloid leukemias with the t(6;11) chromosome translocation. AB - Reciprocal chromosome translocations involving 11q23 are frequently associated with acute leukemias, with the t(4;11) translocation predominating among acute lymphoblastic leukemias, and the t(9;11), t(11;19) and t(6;11) translocations most common among acute myeloid leukemias. In each of these translocations the ALL-1 gene, located at 11q23 and constituting the human homologue of Drosophila trithorax, fuses to a specific gene on the partner chromosome to produce a chimeric protein. Here we report the cloning and the characterization of the partner gene from chromosome 6 (AF-6). AF-6 is expressed in a variety of cell types and encodes a protein of 1612 amino acids. The protein contains short stretches rich in prolines, charged amino acids, serines, or glutamines. In addition, the AF-6 protein contains the GLGF motif shared with several proteins of vertebrates and invertebrates thought to be involved in signal transduction at special cell-cell junctions. PMID- 8242617 TI - Differential activation of cyclophosphamide and ifosphamide by cytochromes P-450 2B and 3A in human liver microsomes. AB - The present study identifies the specific human cytochrome P-450 (CYP) enzymes involved in hydroxylation leading to activation of the anticancer drug cyclophosphamide and its isomeric analogue, ifosphamide. Substantial interindividual variation (4-9-fold) was observed in the hydroxylation of these oxazaphosphorines by a panel of 12 human liver microsomes, and a significant correlation was obtained between these 2 activities (r = 0.85, P < 0.001). Enzyme kinetic analyses revealed that human liver microsomal cyclophosphamide 4 hydroxylation and ifosphamide 4-hydroxylation are best described by a 2-component Michaelis-Menten model composed of both low Km and high Km P-450 4-hydroxylases. To ascertain whether one or more human P-450 enzymes are catalytically competent in activating these oxazaphosphorines, microsomal fractions prepared from a panel of human B-lymphoblastoid cell lines stably transformed with individual P-450 complementary DNAs were assayed in vitro for oxazaphosphorine activation. Expressed CYP2A6, -2B6, -2C8, -2C9, and -3A4 were catalytically competent in hydroxylating cyclophosphamide and ifosphamide. Whereas CYP2C8 and CYP2C9 have the characteristics of low Km oxazaphosphorine 4-hydroxylases, CYP2A6, -2B6, and 3A4 are high Km forms. In contrast, CYP1A1, -1A2, -2D6, and -2E1 did not produce detectable activities. Furthermore, growth of cultured CYP2A6- and CYP2B6 expressing B-lymphoblastoid cells, but not of CYP-negative control cells, was inhibited by cyclophosphamide and ifosphamide as a consequence of prodrug activation to cytotoxic metabolites. Experiments with P-450 form-selective chemical inhibitors and inhibitory anti-P-450 antibodies were then performed to determine the contributions of individual P-450s to the activation of these drugs in human liver microsomes. Orphenadrine (a CYP2B6 inhibitor) and anti-CYP2B IgG inhibited microsomal cyclophosphamide hydroxylation to a greater extent than ifosphamide hydroxylation, consistent with the 8-fold higher activity of complementary DNA-expressed CYP2B6 with cyclophosphamide. In contrast, troleandomycin, a selective inhibitor of CYP3A3 and -3A4, and anti-CYP3A IgG substantially inhibited microsomal ifosphamide hydroxylation but had little or no effect on microsomal cyclophosphamide hydroxylation. By contrast, the CYP2D6 selective inhibitor quinidine did not affect either microsomal activity, while anti-CYP2A antibodies had only a modest inhibitory effect. Overall, the present study establishes that liver microsomal CYP2B and CYP3A preferentially catalyze cyclophosphamide and ifosphamide 4-hydroxylation, respectively, suggesting that liver P-450-inducing agents targeted at these enzymes might be used in cancer patients to enhance drug activation and therapeutic efficacy. PMID- 8242618 TI - Immunohistochemical localization of glutathione S-transferases in human lung. AB - Glutathione S-transferases (GST) detoxify a number of carcinogenic electrophiles including diol-epoxide metabolites of polycyclic aromatic hydrocarbons. The distribution of GSTs A1/A2, M1, M2, M3, and P1 has been studied in lung tissue from 32 subjects by immunohistochemistry using rabbit polyclonal antibodies. GSTA1/A2 and GSTP1 were found to be the most abundant GSTs in human lung, being present in the bronchial and bronchiolar epithelium of all individuals studied. The staining intensity for GSTA1/A2 varied more than that for GSTP1 between individuals. GSTM1, a polymorphic mu-class enzyme, was ambiguously detected in lung tissue and, if expressed, is present at very low levels. GSTM2, a striated muscle-specific isozyme, occurred minimally in the epithelium of the terminal airways, and GSTM3, an enzyme of broad extrahepatic occurrence, was observable in the ciliated airway epithelium and smooth muscle of the lung. The staining for GSTM3 varied from minimal to very intense between individuals; in the bronchial epithelium, it was more abundant in current smokers than in exsmokers. The immunostaining for GSTs in general was most intense in the bronchial epithelium decreasing in the distal airways, in contrast to the previously described peripheral localization of the polycyclic aromatic hydrocarbons activating the P450IA1 enzyme. The localization of GSTs in the bronchial wall suggests that GST polymorphisms may contribute to susceptibility, especially to bronchial tumors of tobacco smokers. PMID- 8242619 TI - A phase I study of the left-shifting agent BW12C79 plus mitomycin C and the effect on the skeletal muscle metabolism using 31P magnetic resonance spectroscopy. AB - BW12C79 stabilizes the oxyhemoglobin molecule resulting in a reversible left shift of the oxygen saturation curve. The activity of a number of bioreductive anticancer drugs, such as mitomycin C, may be enhanced under hypoxic conditions. Twenty-four patients with various malignancies received BW12C79 and mitomycin C. BW12C79 was administered i.v. with a loading dose (20-50 mg/kg) over 1 h followed by a maintenance infusion of 4 mg/kg/h for 5 h. Percentage modification of the oxyhemoglobin (degree of left-shift) was dose related with maximum modification of 56% and was maintained for the duration of maintenance infusion of BW12C79. Hemoglobin electrophoresis showed a fast moving band consistent with the BW12C79 oxyhemoglobin complex. Side effects at the top dose level comprised headache, nausea/vomiting, vein irritation, and myocardial ischemia. One other patient suffered from an acute encephalopathy of unknown etiology a few days following BW12C79. 31P magnetic resonance spectroscopy of exercising calf muscles showed increased breakdown of high energy phosphate stores and a greater reduction in pH. Recovery of the high energy phosphate stores after exercise was slow. These results were consistent with reduced oxygen supply due to either a left shift of the oxygen saturation curve and/or reduced muscle blood flow. BW12C79 did not interfere with the pharmacokinetics of mitomycin C. In conclusion, this phase I study demonstrates the feasibility of achieving a significant left shift in the oxygen saturation curve in cancer patients which is maintained for at least 5 h with acceptable toxicity. The maximum tolerated dose of BW12C79 was 50 mg/kg loading infusion followed by a maintenance infusion of 4 mg/kg/h. Magnetic resonance spectroscopy results were consistent with reduced supply of oxygen to exercising skeletal muscle. BW12C79 may be of potential benefit as an adjunct to bioreductive drugs in the treatment of solid tumors. PMID- 8242620 TI - Evidence for local and systemic activation of immune cells by peritumoral injections of interleukin 2 in patients with advanced squamous cell carcinoma of the head and neck. AB - Interleukin 2 (IL2) was injected peritumorally and intranodally in 36 patients with unresectable squamous cell carcinoma of the head and neck enrolled in an Eastern Cooperative Oncology Group-sponsored phase Ib trial (EST P-Z388). Groups of 6 patients received escalating doses(200, 2 x 10(3), 2 x 10(4), 2 x 10(5), 2 x 10(6), and 4 x 10(6) units) of IL2 daily 5 times/week for 2 weeks. Tumor biopsies were obtained before and after IL2 therapy. Tumor tissue was provided for histology, and the remaining fresh tissue was divided for snap-freezing in -75 degrees C and for separation of tumor-infiltrating lymphocytes (TIL) and tumor cells. Immunophenotyping of TIL performed on cryostat sections of paired pre- and post-IL2 biopsy tissues showed increases after IL2 therapy in the number of T cells (P = 0.005), natural killer (NK; CD16+) cells (P = 0.0001), CD25+ cells (P = 0.004), and HLA-DR+ cells (P = 0.001) accumulating in the tumor stroma. In the tumor parenchyma, NK cells (P = 0.0001) and HLA-DR+ cells (P = 0.003) were increased after IL2 therapy. The T:NK cell ratios in the tumor stroma and parenchyma were decreased after therapy, suggesting selective accumulation of NK cells. By flow cytometry, TIL recovered from post-IL2 biopsy tissues were enriched (P < 0.05) in CD3-CD56+ (NK) cells. In situ hybridization with [35S] cDNA probes for cytokines and IL2 receptors indicated that the numbers of cells expressing mRNA for IL2, tumor necrosis factor alpha, IL1-beta, gamma-interferon, transforming growth factor beta, and IL2 receptor p55 or p70 were increased in post-IL2 biopsy tissues as compared to pre-IL2 tissues. Cytolytic activity of TIL isolated from post-IL2 tissues was also increased, as determined in 4-h 51Cr release assays against K562 targets (12 +/- 3 mean lytic units/10(7) cells +/- SEM pre-IL2 versus 46 +/- 13 post-IL2; n = 16) and against autologous tumor (13 +/- 8 versus 68 +/- 26; n = 9). Fresh TIL of one clinical responder showed relatively high levels (195 lytic units) of autotumor cytotoxicity after IL2 therapy versus no activity prior to therapy. In the blood, NK and lymphokine activated killer cell activity, and percentages of CD3-CD56+ NK cells and of activated (CD25+) T-lymphocytes were increased for all doses of IL2.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8242621 TI - Topoisomerase II inhibition and altered kinetics of formation and repair of nitrosourea and cisplatin-induced DNA interstrand cross-links and cytotoxicity in human glioblastoma cells. AB - By altering the accessibility of DNA sequences for alkylation or platination, and/or for subsequent repair, topoisomerase II can potentially affect the level of DNA interstrand cross-links induced in cells by bifunctional agents. In this study, we investigated the extent to which inhibition of topoisomerase II activity in a human glioblastoma multiforme cell line alters the kinetics of both the formation and the repair of total genomic DNA interstrand cross-links, as well as the sensitivity of the tumor cells to cis-diamminedichloroplatinum II (cis-DDP) and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). Cells were incubated with and without 200 microM novobiocin, a known topoisomerase II inhibitor, for 24 h, followed by exposure to 50 microM BCNU and 25 microM cis-DDP. DNA interstrand cross-linking was determined at various time points over 72 h, using a modified ethidium bromide-DNA binding assay. Sensitivity of the cells to cis DDP and BCNU was also determined with and without novobiocin pretreatment with 200 microM novobiocin. This concentration of novobiocin showed no significant direct cytotoxicity, although it inhibited topoisomerase II activity in tumor cell nuclear extracts by 73%. A significant decrease in the rate of repair of both cis-DDP and BCNU induced DNA interstrand cross-links, with a corresponding decrease in the clonogenic survival of the cells, was observed following novobiocin exposure. Although the peak cross-link indices of novobiocin-treated cells relative to controls were not significantly increased, residual DNA cross linking in the cells after 72 h was increased by 1.4-fold for BCNU and 3-fold for cells treated with cis-DDP, thus, indicating a greater effect of topoisomerase II on cross-link repair than on cross-link formation. These data suggest that inhibition of topoisomerase II may provide a potentially effective clinical strategy for sensitizing human brain tumors, and possibly other tumors as well, to DNA cross-linking anticancer agents. PMID- 8242622 TI - Drug-induced DNA modification in buccal cells of cancer patients receiving carboplatin and cisplatin combination chemotherapy, as determined by an immunocytochemical method: interindividual variation and correlation with disease response. AB - Twenty-six patients with a variety of tumor types were treated according to a phase 1 experimental treatment protocol consisting of repetitive cycles of cis diammine(1,1-cyclobutanedicarboxylato)platinum(II) (carboplatin, 200-480 mg/m2) at day 1 and cis-diamminedichloroplatinum(II) (cisplatin, 50-100 mg/m2) at day 3. Buccal cells were collected in one or two treatment cycles prior to carboplatin, 24 h after carboplatin, just prior to cisplatin, and approximately 24 h after cisplatin administration. Drug-induced DNA modification was visualized at the single cell level by anti-serum NKI-A59 and quantitated by microdensitometry. All (39 of 39) treatments with carboplatin, and almost all (33 of 35) treatments with cisplatin resulted in an increase in nuclear stain. Interindividual variation in drug-induced, adduct-specific nuclear stain amounted to a factor of 5-8 for carboplatin and 5-12 for cisplatin. This drug-induced increase was, however, not related to the dose of either carboplatin or cisplatin, suggesting that large interindividual differences in DNA adduct formation and/or repair obscured the effects of dose variation within the relatively small range used for the drugs (2.4 for carboplatin and 2.0 for cisplatin). This explanation was strengthened by the good reproducibility of the immunocytochemical assay and by the reasonable correlation between carboplatin-induced nuclear stain in cycles 1 and 2 (correlation coefficient, 0.69; P = 0.009). Mean carboplatin-induced nuclear stain was significantly higher in the first cycle than in the second cycle (P = 0.0001) but this difference was no longer significant when drug-induced nuclear stain was corrected for carboplatin dose. Differences in cisplatin-induced nuclear stain between cycle 1 and cycle 2 were small and not significant. Carboplatin-induced nuclear stain was significantly higher in the partial responders than in the nonresponders (P < 0.0001, two cycles combined); the level of statistical significance remained the same after dose correction. Cisplatin induced nuclear stain did not differ significantly between partial responders and nonresponders; this result might, however, be confounded to some extent by remaining carboplatin-induced nuclear stain at the moment of cisplatin administration. It is concluded that determination of the extent of platinum induced DNA modification might be helpful in predicting the tumor response in cancer patients. PMID- 8242623 TI - Therapeutic tumor-specific cell cycle block induced by methionine starvation in vivo. AB - The ability to induce a specific cell cycle block selectively in the tumor could have many uses in chemotherapy. In the present study we have achieved this goal of inducing a tumor-specific cell cycle block in vivo by depriving Yoshida sarcoma-bearing nude mice of dietary methionine. Further, we demonstrate that methionine depletion also causes the tumor to eventually regress. The antitumor effect of methionine depletion resulted in the extended survival of the tumor bearing mice. The mice on the methionine-deprived diets maintained their body weight for the time period studied, indicating that tumor regression was not a function of body weight loss. The data reported here support future experiments utilizing methionine depletion as a target for tumor-selective cell cycle dependent therapy. PMID- 8242624 TI - Pharmacokinetics and bioactivity of 1,4,7,10-tetra-azacylododecane off',N'',N''' tetraacetic acid (DOTA)-bismuth-conjugated anti-Tac antibody for alpha-emitter (212Bi) therapy. AB - A major factor that is critical to the potential effectiveness of alpha-emitter 212Bi radioimmunotherapy is the design of radiometal-chelated antibodies that will be stable in vivo. The chelate should bind the radiometal firmly to minimize release of the radionuclide from the monoclonal antibody-chelate complex. The present study examines a member of a new class of polyamine carboxylate chelating compounds, the DOTA ligands, for conjugating radiometal ions to antibody. Biocompatibility and stability are assessed with the anti-Tac monoclonal antibody that is directed against the human interleukin 2 receptors. The scientific basis for the clinical use of this antibody in radioimmunotherapy is that resting normal cells do not express the interleukin 2 receptor, whereas the receptor is expressed on the surface of certain neoplasms and by activated T-cells in select autoimmune diseases and in allograft rejection. First, we examined the impact of the labeling procedure and the presence of the chelate, DOTA, on antibody bioavailability and survival. Next, we studied the capacity of the antibody chelate complex to retain radiobismuth. Coupling DOTA to antibody or adding Bi(III) to DOTA-coupled antibody did not disturb antibody immunoreactivity in in vitro binding studies. In addition, as analyzed by in vivo studies, DOTA-antibody dummy labeled with nonradioactive bismuth showed pharmacokinetics and tissue distribution identical to those of antibody not modified with DOTA. DOTA-anti-Tac charged with radioactive bismuth showed pharmacokinetics identical to radioiodinated dummy-labeled DOTA-antibody, suggesting little premature release of radioactive bismuth from the antibody complex. Moreover, in the early, therapeutically relevant time points (2 h and 6 h), there was no significant preferential accumulation of bismuth in any organ. At the 5-day time point, beyond the range of therapeutic interest, there was delayed excretion of bismuth from reticuloendothelial tissues relative to radioiodine from catabolized antibody. Excretion of catabolized DOTA-bismuth had an apparent t1/2 of approximately 1 day without the marked renal accumulation typical of the free bismuth ion. The compatibility of DOTA conjugation with antibody bioactivity and the stability of the radioactive bismuth complex in vivo provide important preclinical validation of the potential utility of this new chelating agent for 212Bi monoclonal antibody radioimmunotherapy in humans. PMID- 8242625 TI - In vivo mutagenesis induced by CC-1065 and adozelesin DNA alkylation in a transgenic mouse model. AB - Although considerable work has focused on characterizing the bonding chemistry and sequence selective alkylation of DNA by cyclopropylpyrroloindole compounds, little is known about the molecular consequence of their N-3-adenine adducts in whole animal systems. We have utilized a transgenic mouse system, harboring a lambda phage shuttle vector, to assess the mutagenic potential of the antitumor compounds CC-1065 and adozelesin and, for the first time, to track the in vivo fate of their unique DNA modifications at the nucleotide level. Mice were inoculated with a single therapeutic dose of these agents and sacrificed at either 18 h, 3 days, or 15 days for extraction and analysis of liver DNA. Mutant frequencies obtained from drug treated and control animals were determined by in vitro packaging of the phage vector from genomic DNA followed by a colorimetric plaque assay to screen for phage in which the accompanying lacI repressor gene had mutated. Although undetectable at 18 h posttreatment, by 72 h a 3-fold increase in mutant frequency was observed in drug treated animals such that sequence analysis of drug induced mutations could be performed and a direct comparison made between in vitro and in vivo DNA alkylation. Base substitution involving guanine or cytosine accounted for 64% of the 41 mutations sequenced from drug treated animals. Only 7 of the mutations occurred at a cyclopropylpyrroloindole alkylation site while 23 occurred 1 to 4 nucleotides from a potentially alkylated adenine. PMID- 8242626 TI - Cytotoxicity of antifolate inhibitors of thymidylate and purine synthesis to WiDr colonic carcinoma cells. AB - We have studied the cytotoxicity of 5,10-dideazatetrahydrofolate (DDATHF) and of D-1694 to human WiDr colonic carcinoma cells as a model system for the effects of pure inhibitors of either the de novo purine synthesis pathway or thymidylate synthesis. The growth of this cell line was inhibited by very low concentrations of either agent and the lethality of DDATHF and D-1694 was completely prevented by continuous exposure to either hypoxanthine or thymidine, respectively, indicating that these compounds were very potent metabolic inhibitors, each specific for one of these pathways. D-1694 was highly cytotoxic (> 3 logs of kill) after a 4-h exposure to 1 microM drug, or a 24-h exposure to very low concentrations (0.04 microM). On the other hand, the cytotoxicity of DDATHF was substantially lower, with 2 logs of cell kill requiring >> 100 microM with 4 h of exposure or approximately 40 microM for 72 h of exposure. Maximal cell kill induced by D-1694 was 5-6 logs, consistent with elimination of all viable cells except preexisting mutants. A maximum of 2-3 logs of cell kill was observed with DDATHF. Exposure of WiDr cells to either D-1694 or DDATHF caused striking cellular changes, but the morphologies of cells treated with the two drugs were remarkably different. D-1694-treated cells detached from the dish within 1-2 days after a megaloblastosis, whereas DDATHF-treated cells remained adherent to the dishes for at least 10 days after treatment. The addition of thymidine to D-1694 treated cultures or hypoxanthine to DDATHF-treated cells after up to 20 h of drug exposure completely prevented cytotoxicity of either drug. With longer exposures, cytotoxicity of both drugs progressively increased in spite of such rescue. Our results indicate that substantial (99-99.9%) tumor cell kill can be induced by a pure inhibitor of purine synthesis, but that the rate of commitment to cell death and the extent of cell kill is greater with a pure inhibitor of thymidylate synthesis. PMID- 8242627 TI - cis-diamminedichloroplatinum(ii) resistance in vitro and in vivo in human embryonal carcinoma cells. AB - In the embryonal carcinoma cell line Tera and its 3.7-fold cis diamminedichloroplatinum(II) (CDDP)-resistant subline, Tera-CP, parameters were studied that might have changed in relation to induction of CDDP resistance. Phenotypes of both lines were embryonal carcinoma. Karyotypes were related with a decreased mean number of chromosomes and fewer copies of the short arm of chromosome 12 in Tera-CP. Tera-CP showed cross-resistance for melphalan and 4 hydroperoxycyclophosphamide and had an 1.4-fold increased glutathione (GSH) level, a 1.5-fold increased glutathione S-transferase (GST) activity, and a 1.4 fold increased GST pi expression compared to Tera. Tera-CP was cross-resistant to 5-fluorouracil, but thymidylate synthase activity was not increased. Topoisomerase I and II activities and c-myc RNA and protein expression were the same in both lines. Platinum accumulation was equal in both lines, and platinum DNA binding was lower in Tera-CP than in Tera. Both cell lines were xenografted into nude mice and tumors showed marked differentiation. Tera-CP tumors were 2.8 fold resistant to CDDP compared to Tera tumors. In new cell lines derived from xenografts of Tera and Tera-CP CDDP sensitivity, GST activity and GSH level corresponded with their sensitivity and resistant origin. Tera-CP is a model of in vitro and in vivo CDDP resistance with the GSH/GST detoxifying system as an important mechanism. CDDP resistance could be induced without a concomitant increase in differentiation. PMID- 8242628 TI - Detection of hypoxic cells by monoclonal antibody recognizing 2-nitroimidazole adducts. AB - Hypoxic cells in tissue pose many medical problems, and there is a need for more accurate measurements of tissue hypoxia. However, measurement of the pO2 and the extent of hypoxia within normal and tumor tissue have proven difficult. One of the most sensitive of the currently available methodologies involves the oxygen dependent metabolic activation of nitroheterocyclic drugs, leading to adducts between the drugs and cellular macromolecules. Limitations of the present drugs and adduct-detection methods prompted the present studies. A pentafluorinated derivative [EF5; 2-(2-nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3 pentafluoropropyl)acetam ide] of etanidazole was synthesized with the expectation of lessening some of the non-oxygen-dependent variability in adduct formation observed previously with other nitroaromatic compounds. EF5-protein conjugates, prepared by radiochemical reduction, were found to be immunogenic and allowed the development of monoclonal antibodies. One of these antibodies, ELK2-4, has been characterized and found to be highly specific for the EF5 adducts whether produced radiochemically or by cellular bioreductive metabolism. 9L rat glioma cells pretreated with EF5 under hypoxic, compared with aerobic, conditions were readily discriminated immunochemically using fluorochrome-conjugated secondary antibodies which recognize the ELK2-4 antibody subtype (IgG1). Similarly, the central region of multicenter spheroids, composed of EMT6 mouse mammary sarcoma cells, was selectively visualized by immunohistochemistry after the spheroids were incubated for 4 h in 0.5 mM EF5. Tumor biopsy, preparation, and immunohistochemical staining 24 h after treatment of tumor-bearing animals with drug also demonstrated high contrast regions within EMT6 mouse or Morris 7777 hepatoma rat tumors. The use of this new compound and its highly specific monoclonal antibody may allow elucidation of bioreductive metabolism of the nitroheterocyclics and significantly improve technologies for the quantitation of tissue pO2. PMID- 8242629 TI - Preclinical evaluation of 67Cu-labeled intact and fragmented anti-colon carcinoma monoclonal antibody MAb35. AB - The anti-carcinoembryonic antigen murine monoclonal antibody MAb35 and its F(ab')2 fragment were labeled with 131I or the potential therapeutic nuclide 67Cu. In vivo distribution patterns were compared in nude mice bearing human tumor xenografts by coinjection of the 131I- and 67Cu-labeled materials, thereby minimizing variations due to xenograft and host animal. The results showed that the 67Cu-labeled intact MAb35 achieved twice the percentage of injected dose/g tumor when compared to its 131I-labeled counterpart, without significant impairment of the wholebody distribution pattern. However, this effect was not evident in the case of F(ab')2, where high uptake of 67Cu was found in the kidney without any enhancement of accumulation in the target xenografts. To investigate the underlying causes of the different distribution patterns observed, iodine labeling was also performed using a more stable linkage, and the results indicated that the observed differences cannot be explained by simple deiodination of conventionally labeled preparations. We conclude that the intact form of the 67Cu-labeled antibody may be superior to the F(ab')2 fragment for use in our intended clinical studies. Our continuing work on the processing of radiometal-labeled F(ab')2 fragments, at the systemic and cellular level, will hopefully lead to a strategy to circumvent the problem of high kidney accumulation. PMID- 8242630 TI - The use of daunomycin-antibody immunoconjugates in managing soft tissue sarcomas: nude mouse xenograft model. AB - Analysis of human fibrosarcoma cells exposed to radiolabeled monoclonal antibody 19-24, which recognizes sarcoma-associated antigen p102, revealed that over 54% of the cell surface-bound radioactivity was internalized. No modulation of cell surface p102 antigen by monoclonal antibody 19-24 was observed in human fibrosarcoma cells. Monoclonal antibody 19-24 coupled to daunomycin via a dextran bridge was found to be most effective. In different preparations, the daunomycin:total protein molar ratio ranged from 1.9 to 6.1. In vitro cytotoxicity studies using human fibrosarcoma cells showed that, at 10 micrograms/ml concentration, this immunoconjugate was 79.4% as efficient as free daunomycin and, at 1 microgram/ml concentration, 36.8% as efficient. Control nonspecific murine monoclonal antibody P3 immunoconjugates were relatively ineffective. The distribution of 14C-Adriamycin, 125I-labeled monoclonal antibody 19-24, and 125I-labeled 19-24 immunoconjugate was also evaluated over a 24-h period in tumor and normal tissues of athymic mice bearing a human fibrosarcoma xenograft. Poor uptake of radiolabeled Adriamycin by the tumor tissue was observed. The level of 14C radioactivity in the tumor tissue never exceeded 1% of the total injected dose and was 24.8-fold lower than the radioactivity found in the spleen tissue. Tumor tissue uptake of radiolabeled monoclonal antibody 19-24 was characterized by the high tumor tissue:blood ratio of 1.62 +/- 0.28 (SD). However, for monoclonal antibody 19-24 immunoconjugates, this ratio decreased to 0.66 +/- 0.05, which was still higher than normal (liver, 0.48 +/- 0.02; lung, 0.48 +/- 0.07; spleen, 0.28 +/- 0.01) or nonspecific monoclonal antibody P3 immunoconjugates (0.22 +/- 0.03). Thus, it appears that, compared to free daunomycin, monoclonal antibody 19-24 immunoconjugates may be more efficient and less cytotoxic to normal tissues. PMID- 8242631 TI - TP53 gene mutation profile in esophageal squamous cell carcinomas. AB - Esophageal squamous cell carcinoma is a form of cancer occurring most commonly in males, particularly those living in some areas of Asia, Africa, and western Europe. In some of these tumors, a sequence alteration has been identified in the coding region of the TP53 gene which is known to inactivate the tumor suppressor function of its product. Using a GC clamp (i.e., a GC rich domain) denaturing gradient gel electrophoresis assay we have been able to identify sequence modifications in 27 of the 32 tumor samples analyzed (84%). Most of the mutations occur in exon 6, a region of the gene which has not previously been reported as being a hot spot for the mutations of other cancers. Twelve of the mutations reported here have not been described in other types of tumors and these consist mostly of frameshift or splice mutations. The distribution of mutations [transitions (45%), transversions (34%), and frameshift (21%)] suggests that the etiological contribution of genotoxic factors might be complex and might associate different exogenous and endogenous mutagen exposures. PMID- 8242632 TI - Mutagenicity induced by hyperthermia, hot mate infusion, and hot caffeine in Saccharomyces cerevisiae. AB - Mate, an infusion containing caffeine (3 g/liter), is drunk hot by most Uruguayan, North Argentinian, and South Brazilian people. This beverage has been recently associated with esophageal cancer in Brazil and Uruguay. To test the mutagenic and lethal effects of mate infusion, caffeine, hyperthermia, and their combinations, we used Saccharomyces cerevisiae as an eucaryotic model system measuring lys to LYS reversions. We showed that mate infusion was not mutagenic, whereas caffeine at the same concentration contained in mate, produced a 5-fold increase in the spontaneous mutation rate. The highest observed mutagenic rate corresponded to hyperthermia (54 degrees C at 60 min). Hot caffeine also produced a time-dependent mutagenic effect, whereas hot mate infusion determined a significantly lower mutagenic effect than hot caffeine. The differential lethality produced by the tested agents plays an important role in the expression of the induced mutagenic damage. Caffeine and mate infusion could decrease the mutagenic effect of hyperthermia through the channeling of part of the induced DNA lesions into an error-free repair pathway. PMID- 8242633 TI - Extinction of expression of the PU.1/Sfpi-1 putative oncogene encoding a B-cell- and macrophage-specific transcription factor in somatic cell hybrids. AB - Several examples of extinction of cell type-specific gene expression have been observed following fusion of different cell types. Possible mechanisms of the extinction include loss of transcriptional activators and acquisition of repressor factors responsible for cell type-specific gene expression. In this study, we demonstrated the extinction of expression of the PU.1/Sfpi-1 putative oncogene encoding a B-cell- and macrophage-specific transcription factor when plasmacytoma cells are fused with embryonal carcinoma (EC) cells. The hybrid cells retained most chromosome complements from both parental lines including chromosome 2 on which the PU.1 gene is located. Therefore, extinction of PU.1 gene expression in the hybrids is not likely the result of chromosome segregation but rather due to a transacting negative factor(s) present in EC cells. On the contrary, expression of the PU.1 mRNA in plasmacytoma cells was not extinguished upon cell fusion with T-lymphoma cells, although the parental T-lymphoma cells did not express PU.1 transcripts. Hence, T-lymphoma cells seemed to be permissive to PU.1 gene expression, while EC cells were repressive. These results suggest that PU.1 gene expression which positively regulates some B cell- and macrophage specific gene expression is a target of negative regulatory mechanisms during cell differentiation, and the regulatory mechanisms repressing PU.1 gene expression is different between EC cells and T-cells. PMID- 8242634 TI - Cloning and characterization of overexpressed genes in the mammary gland of rat strains carrying the mammary carcinoma suppressor (Mcs) gene. AB - Inbred rat strains vary in their susceptibilities to mammary carcinogenesis. The Copenhagen (COP) and Wistar-Kyoto (WKY) rats are tumor resistant, whereas the Wistar-Furth (WF), Fischer (F344), and outbred Sprague-Dawley (SD) rats are susceptible. A dominant pattern of inheritance acting via the mammary carcinoma suppressor (Mcs) gene(s), which is mainly responsible for mammary tumor resistance, has been defined in the COP and WKY rats. In order to understand the basis of the phenotype, COP and WF mammary mRNAs were used for subtractive hybridization to isolate genes associated with the activity of the Mcs gene(s). Three genes, alpha-casein, lipoprotein lipase, and an unidentified gene, were found to be overexpressed in the mammary gland of the COP rat. In addition to alpha-casein overexpression, Northern analysis demonstrated that beta- and gamma casein genes were also highly expressed in the mammary glands of tumor-resistant WKY and COP virgin rats but not the susceptible F344, WF, and SD strains. The association of casein gene expression with the tumor-resistant phenotype was further investigated by determining the functional site of the strain-specific casein gene regulation by using a mammary cell transplantation assay. In contrast to its normal endocrine control during pregnancy and lactation, casein gene overexpression was found to be controlled within the mammary epithelial cells of virgin rats. This is also the site of production and action of the Mcs gene product. Comparison of polymerase chain reaction-amplified beta-casein precursor RNA levels with the use of reverse transcription-polymerase chain reaction revealed that the regulation of this gene is likely at the transcriptional level. These data suggest an association of overexpression of casein genes, with the Mcs phenotype. The biological significance of this association is under investigation. PMID- 8242635 TI - Three discrete regions of deletion at 3p in head and neck cancers. AB - Alteration of the short arm of chromosome 3 is one of the most consistent cytogenetic abnormalities found in human head and neck cancers. These alterations, composed of translocations and deletions, have been associated with the presence of a tumor suppressor gene(s), but no clear evidence of the location of this presumptive gene(s) was available. We performed a molecular analysis of the 3p region using a polymerase chain reaction-based approach. Twenty-eight of the 38 cases analyzed (74%) showed the presence of single or multiple areas of allelic loss. Three commonly deleted regions, tentatively mapped to 3p24-ter, 3p21.3, and 3p14--cen, were identified. Our results suggest that at least three oncosuppressor genes mapping on 3p may be involved in head and neck cancer development and support a common oncogenic pathway with squamous cell lung cancer, for which a similar pattern of 3p deletion has been described recently. PMID- 8242636 TI - Allelic deletions in renal tumors: histopathological correlations. AB - Allelic loss on the short arm of chromosome 3 (3p) is considered to be one of the early detectable events in the pathogenesis of renal cell carcinoma (RCC). Conflicting reports, however, suggest that this event may be absent in some renal tumors. The present study attempts to further define subgroups of renal tumors associated with 3p deletions. In addition, we have also attempted to identify late genetic events associated with tumorigenesis and tumor progression. Eighty two primary renal tumors (69 RCC and 13 oncocytic tumors) were analyzed by restriction fragment length polymorphism analysis directed at chromosomes 3, 11p, 17p, and 18q. Results were correlated with histopathological information. Deletions of 3p were seen in nonpapillary RCC of all cell types, but were absent in oncocytic and most papillary tumors. Among the 60 nonpapillary RCC, significant correlations were seen between deletion of 17p and tumor grade (P = 0.037), P stage (P = 0.027), and nodal metastases (P = 0.042). We therefore conclude that 3p deletions, although not specific to any cell type or histological pattern of RCC, are seen in a majority of clear cell nonpapillary RCC but are absent in oncocytic and most papillary tumors. Additional allelic losses on chromosome 17p are associated with advanced disease and, therefore, may be related to tumor progression. Further studies on larger series of patients with extended follow-up will be necessary to investigate the prognostic value of molecular genetic markers in RCC. PMID- 8242637 TI - Gene transfection and expression of the ovarian carcinoma marker folate binding protein on NIH/3T3 cells increases cell growth in vitro and in vivo. AB - The glycoprotein gp38 is overexpressed in 90% of ovarian carcinomas and recognized by monoclonal antibodies MOv18 and MOv19. This molecule is a high affinity folate binding protein (FBP) and a potential marker for ovarian carcinoma. We have developed a model to investigate the biochemical and biological properties of this folate receptor by transfecting NIH/3T3 cells, which do not endogenously express FBP, with a vector containing the complementary DNA for the gp38 cloned from the ovarian carcinoma cell line IGROV1. The FBP expressed shows features identical to those of the protein produced by IGROV1 cell. The FBP is expressed on the cell membrane in a glycosylphosphatidylinositol linked form, since it is released by treatment with phosphatidylinositol-specific phospholipase C, and is shed into the culture medium of the NIH/3T3 transfectants. Immunoblot analysis with MAbs MOv18 and MOv19 showed that both the glycosylphosphatidylinositol-linked and the soluble FBP migrate at the same apparent molecular weight as the respective IGROV1 proteins. The FBP-transfected NIH/3T3 cells bound folic acid and internalized about 30-fold more folic acid than mock-transfected cells. Growth analysis revealed that FBP-transfected NIH/3T3 cells like IGROV1 maintained their growth rate after 10 days of culture in medium containing physiological or low folate concentration, and tumors arising after transplanting FBP-tNIH/3T3 cells in nude mice were 3-fold heavier than those arising after transplantation of non-FBP-expressing NIH/3T3 cells. These results suggest a correlation between human ovarian carcinoma growth and FBP overexpression. PMID- 8242638 TI - Infrequent mutations of the p53 gene in pulmonary carcinoid tumors. AB - Archival specimens of 25 pulmonary carcinoids including 15 cases of typical carcinoid, 9 atypical carcinoids, and 1 large-cell neuroendocrine carcinoma were analyzed for mutations in exons 5 to 8 of the p53 gene. Mutations were identified in 4 tumors, including 3 out of 15 (20%) typical carcinoids and the single large cell neuroendocrine carcinoma, but none of the atypical carcinomas showed a mutation. The mutations were acquired during tumor development since they were not present in the corresponding nontumorous tissue. All mutations in the typical carcinoids, a tumor type without epidemiological link to cigarette smoking, were G to A transitions. The level of p53 protein was investigated by immunohistochemistry with the polyclonal antibody CM-1. None of the pulmonary carcinoids investigated showed a positive reaction, despite the presence of missense mutations in two cases. Negative staining of carcinoids with mutations was also observed with the monoclonal antibodies pAb1801 and DO-1. Our data suggest that point mutations of the p53 gene are infrequent in pulmonary carcinoids thus contrasting the findings in other histological types of lung cancer, in particular small-cell lung cancer. Moreover, negative immunostaining for p53 is no indicator for the absence of p53 missense mutations in typical carcinoids. PMID- 8242639 TI - Serum and plasma M(r) 92,000 progelatinase levels correlate with spontaneous metastasis of rat 13762NF mammary adenocarcinoma. AB - The expression of metalloproteinases, such as type IV collagenase/gelatinase, enables tumor cells to degrade type IV collagen present in the basement membrane and correlates with metastatic potential of several tumor types. We found that increased levels of rat serum type IV collagenolytic activity are associated with increased 13762NF mammary adenocarcinoma metastases in lungs and lymph nodes of syngeneic rats. To investigate serum metalloproteinases responsible for type IV collagenolysis, we performed zymography and Western blot analysis of rat sera. A M(r) 92,000 progelatinase (progelatinase B, M(r) 92,000 type IV procollagenase, MMP-9) was detected on zymograms of rat sera within 16 days after intramammary fat pad inoculation of highly metastatic MTLn3 cells. The activated serum M(r) 92,000 progelatinase degraded sodium dodecyl sulfate-denatured type I and IV collagens but was not active on casein, albumin, hemoglobin, and immunoglobulin. Sera from rats with fat pad tumors and lung metastases formed from highly metastatic clones possessed greater than 7 times higher levels of serum M(r) 92,000 progelatinase than control sera or sera from rats bearing similar size fat pad tumors of low or nonmetastatic clones. The results were confirmed by Western blot analysis of rat sera using rabbit anti-human M(r) 92,000 progelatinase antibodies. Similar results were obtained from the analysis of rat plasma samples. The serum and plasma M(r) 92,000 progelatinase levels correlated with the extent of metastases in the lung and lymph nodes. The results indicate that high levels of serum and plasma M(r) 92,000 progelatinase are associated with the presence of disseminated metastatic adenocarcinoma cells and suggest that this enzyme may facilitate the invasion of blood-borne tumor cells through vascular basement membranes. PMID- 8242640 TI - Quantitative differential effects of rhodamine 123 on normal cells and human colon cancer cells by magnetic resonance spectroscopy. AB - Rhodamine 123 is a lipophilic cationic compound that is selectively taken up by cancer cell mitochondria. This compound is toxic to epithelial cancer cells in vitro and displays significant anticancer activity in vivo. However, the mechanism of action of rhodamine 123 in intact, actively metabolizing cell preparations is unknown. We have used 31P- and 13C-nuclear magnetic resonance spectroscopy to quantitatively characterize how rhodamine 123 affects the energetics of human colon cancer cells (HCT-116) and spontaneously immortalized normal epithelial cells (CV-1). Rhodamine 123 differentially altered the phosphorus and glucose metabolism of HCT-116 and CV-1 cells. 31P-nuclear magnetic resonance detected mitochondrial poisoning in the HCT-116 human colon cancer cell line in its early stages after selective uptake of rhodamine 123. When we compared administration of rhodamine 123 and [1-C13]glucose to administration of [1-C13]glucose alone in the HCT-116 cells, we noted a marked decrease in intracellular pH to 6.7 +/- 0.06 (mean +/- SD) units, a 2.2-fold increase in lactate production, and a 1.8-fold increase in glucose consumption after 10 h. In addition, we found a 2-fold rise in intracellular free magnesium 12 h after rhodamine 123 administration. These results suggest that when rhodamine 123 inhibits mitochondrial ATP production, it initially stimulates cytoplasmic glycolysis in an attempt to maintain cellular energy demands. The marked fall in intracellular pH and rise in intracellular free magnesium after administration of rhodamine 123 may inhibit activity of several glycolytic enzymes: this effect would inhibit cytoplasmic ATP generation and interfere with multiple cell enzymatic processes, leading to cell death. The CV-1 cells showed no change in intracellular pH, intracellular free magnesium, or magnesium-bound ATP levels over the 24-h period following rhodamine 123 administration. Rhodamine 123 also failed to alter glucose utilization and lactate production levels significantly in the CV-1 cells. These results prove the usefulness of 31P- and 13C-nuclear magnetic resonance spectroscopy for quantifying differing effects of rhodamine 123 on the high energy phosphate metabolism and glucose metabolism of HCT-116 and CV-1 cells. PMID- 8242641 TI - Antiproliferative effects of isoflavones on human cancer cell lines established from the gastrointestinal tract. AB - Seven isoflavones, biochanin A, daidzein, genistein, genistin, prunectin, puerarin, and pseudobaptigenin were tested for cytostatic and cytotoxic effects on 10 newly established cancer cell lines of the human gastrointestinal origin. Proliferation of HSC-41E6, HSC-45M2, and SH101-P4 stomach cancer cell lines was strongly inhibited by biochanin A and genistein, whereas other stomach, esophageal, and colon cancer lines were moderately suppressed by both compounds. Biochanin A and genistein were cytostatic at low concentrations (< 20 micrograms/ml for biochanin A, < 10 micrograms/ml for genistein) and were cytotoxic at higher concentrations (> 40 micrograms/ml for biochanin A, > 20 micrograms/ml for genistein). DNA fragmentation was observed at cytotoxic doses of both compounds, indicating the apoptotic mode of cell death by the compounds. Chromatin condensation and nuclear fragmentation of each cell line were also observed. The advent of apoptosis was dose dependent for both isoflavones. Biochanin A suppressed tumor growth of HSC-45M2 and HSC-41E6 lines in athymic nude mice. Our results suggest that two of isoflavone derivatives, biochanin A and genistein, inhibit the cell growth of stomach cancer cell lines in vitro through activation of a signal transduction pathway for apoptosis. Moreover, in vivo experiments demonstrate that biochanin A can be used as an anticancer agent. PMID- 8242642 TI - Correspondence re: J. P. Alexander et al., T-cells infiltrating renal cell carcinoma display a poor proliferative response even though they can produce interleukin 2 and express interleukin 2 receptors. Cancer Res., 53: 1380-1387, 1993. PMID- 8242644 TI - 60 years of the Institute of Oncology in Warsaw. PMID- 8242645 TI - Cancer in Poland. AB - As in other European countries, cancer is the second most frequent cause of death in Poland. Estimates say that some 100,000 Poles will have developed various cancers every year by the end of the 1980s. The number of cancer-related deaths reached 71,509 by 1989. The constantly growing number of new cases and deaths from cancer is one of the most characteristic features of malignant neoplasms in Poland. The recent decades have seen major changes in the structure of cancer in Poland. This is true for both sexes. By the end of the 1980s, the predominating neoplastic disease in men was cancer of the lung. It was the cause of 34.3% of cancer-related deaths in 1989. In women, malignant neoplasms of the breast, large bowel, and stomach represent about 33% of all cancer localizations. Geographic distribution of mortality ascribed to cancer shows an east-west pattern. This is true particularly for such cancer sites as the large bowel, gall bladder, pancreas, uterine cervix, urinary bladder, breast, ovary; it seems to have a clear connection with the geographic pattern of urbanization in Poland. PMID- 8242646 TI - Survival of cancer patients in Poland. AB - Data on survivals that were obtained from population-based cancer registries and representing different subpopulations in Poland, i.e., the City of Warsaw and the Warsaw Rural Areas (WRA), are presented. The overall 5-year relative survival rates calculated for males increased from 23.8% in 1963-1968 to 31.2% in 1978 1981, while no such increase was found among females (43.6 and 43.2%, respectively). The 5-year survivals were divided into three groups: (1) fairly good (40 to 100%); (2) moderate (20 to 39%); and (3) poor (below 20%). The first group also included (apart from cancer of the skin and lip, where the 5-year survival rate was 100%) cancers of the corpus uteri, Hodgkin's disease, lymphatic leukemia, malignant melanoma, testis, larynx, breast, thyroid, cervix, uteri, and prostate. The second group included cancers of the colon, bone, ovary, rectum, urinary bladder, kidney, non-Hodgkin's lymphoma, and brain. The lowest-survival group included, among other localizations, cancers of the lung, stomach, gall bladder, pancreas, esophagus, and liver. A comparison between the 5-year survivals in Warsaw City, WRA, and the rates obtained in Finland and the U.S. showed that Polish results are considerably worse than in the other countries. PMID- 8242647 TI - Survival studies: Cracow Cancer Registry experience. AB - The experience of the Cracow Cancer Registry is presented to show how useful a population-based survival study can be. Survival rates for lung, breast, and colorectal cancer are compared over two periods of time. In addition, the survival experience of patients treated at the Oncologic Center in Cracow is compared with the U.S. SEER data. PMID- 8242648 TI - Tobacco, alcohol, and diet in cancer etiology in Poland: a review of Polish epidemiological studies. AB - A review of the most important case-control studies conducted in Poland and published over the last 30 years, which indicate a role of tobacco, alcohol, and diet in the etiology of cancer, are presented. Very little was done in Poland to publicize these results. The current discussion on causes of the high incidence and mortality from cancer in Poland is now focusing on the extremely high environmental pollution in Poland rather than on such lifestyle hazards as tobacco smoking, alcohol drinking, and poor dietary habits. All studies presented here were performed in the Silesian and Cracow Regions of Poland, which are considered to be the most polluted areas. Awareness of the danger of pollution is very high among the inhabitants and the decision makers; however, awareness of the dangers of one's lifestyle is still relatively low and has been discussed in terms of disbelief rather than in terms of any well-defined facts that should lead to proper priorities for public health action. PMID- 8242649 TI - Trends in the tobacco-related cancer epidemic in Europe. AB - Recent trends in mortality from lung cancer and other tobacco-related neoplasms in Europe are reviewed. During the last decade, overall lung cancer mortality in males showed no systematic pattern in northern and central Europe, but some modest decline started at younger ages in several countries. In southern Europe, lung cancer mortality started from lower values, but is still rising, and only in Italy is some flattening of rates at relatively high levels becoming apparent in middle age (35 to 64 years). The average change in lung cancer rates in southern Europe over the last decade for males was +24% at all ages and +22% in middle age. The upward trends were even more substantial in eastern European countries (+32% in middle age), which now have the highest lung cancer rates in young and middle-aged males. Over the last few decades, female lung cancer rates have risen in all European countries, but only in Denmark and Britain are overall rates now over 20/100,000. There is therefore still ample scope for urgent intervention aimed at controlling a major tobacco-related lung cancer epidemic among European women in the near future. Substantial rises have been observed for cancer of the oral cavity and pharynx in most European countries, and for cancers of the esophagus, larynx, and (to a lesser extent) of the bladder in eastern Europe. Thus, southern and mainly eastern Europe are becoming priority areas for a campaign for giving up smoking since the prevalence of tobacco smoking in the young is higher and high-tar dark-tobacco cigarettes are still common. PMID- 8242650 TI - The United States National Cancer Institute and the science of tobacco control research. AB - U.S. efforts to control tobacco use and tobacco-related morbidity and mortality have been reasonably successful over the past 3 decades, during which there has been a 34% reduction in adult smoking. Nevertheless, tobacco use remains a significant public health problem in the U.S., with more than 430,000 tobacco related deaths per year and over one fourth of the population continuing to smoke. Many organizations are involved in tobacco-use control activities, the most broadly focused of which is that of the National Cancer Institute (NCI). The NCI's program is described and its emphasis on a data-based decision matrix in its approach to tobacco and cancer control research and applications of research is discussed. Finally, future approaches to tobacco-use control in the U.S. are suggested. PMID- 8242651 TI - Potential contribution of screening to cancer mortality reduction. AB - The objective of screening for cancer is to reduce mortality or to improve quality of life. Screening is practiced for several anatomical sites and by several tests. Only cervical cancer screening based on cytological smears has been shown to be effective as a public health policy. Screening for breast cancer based on mammography was shown to reduce mortality in several randomized trials and nonexperimental studies. However, no data are available on its effectiveness at population levels in terms of a public health policy. There are several other valid tests. Application of these tests has failed to demonstrate a reduction in mortality or such an application was never tried and tested. This emphasizes that the prerequisite for effective screening is that the total program is valid and not only the test. At present, knowledge on screening is not fully applied in most European countries. Regular screening for cervical cancer will result in a 90% reduction in the risk of invasive disease. It is likely that the protective effect for breast cancer is about 30%. At the population level, smaller protections are to be expected, and well-organized screening programs are likely to reduce the total cancer death rate by 6 to 10% among females. PMID- 8242652 TI - The European action against cancer. AB - 1. Prevention. In this field the main goal is the fight against tobacco. The following measures have been proposed and are now being implemented within the European community: banning smoking in places open to the public, regulation of maximum tar content of cigarettes, health warnings on all tobacco products, ban of oral tobacco, harmonizing tobacco prices upward. The banning of direct and indirect advertising has been proposed but has not yet been accepted. 2. Screening. Pilot projects on breast cancers and cervix cancer screening were launched in all member states. Guidelines for quality assurance were prepared and circulated for both types of screening. Exchange of information was organized through a networking of pilot studies. 3. Informing the public. A European code against cancer was widely distributed with the help of associations, cancer leagues, and practitioners. Health education at school (from 5 to 12 years of age) was recommended, a program was adopted, and pilot projects for the education of teachers were supported. 4. Education and training of health professionals. Emphasis has been placed on general practitioners whose role is crucial. 5. Research. A wide prospective study on diet and cancer was organized in six member states. A cohort of over 500,000 individuals will be monitored, follow-up, whose samples of urine and blood will be kept in liquid nitrogen. Clinical research is supported. In conclusion, despite a relatively small budget (about 11 million $U.S. per year), it was possible to support and coordinate several actions and therefore to stimulate the fight against cancer in the European community. PMID- 8242653 TI - Potential contribution of therapy to cancer mortality reduction. AB - Reductions in cancer mortality may come about for a number of reasons, including improvements in treatment. The impact will vary from cancer to cancer. For some, expert curative surgery is crucial, whereas for others, the use of appropriate chemotherapy is a key factor. Examples of the latter, in which there are already discernible reductions in national cancer mortality data resulting from chemotherapy, include testicular cancer and Hodgkin's disease. For more common diseases, such as ovarian cancer, reductions also are being seen. For others, such as breast and colorectal cancer, the current more widespread use of adjuvant chemotherapy may lead to overall mortality reduction in the future. It should be recognized that chemotherapy should be given only by those experienced in its use, and that this facility should form part of a larger provision for health care in relation to cancer, ranging from public education to population screening and from better oncology training for clinicians to greater encouragement to participation in clinical trials. New drug development is clearly a priority, but further advances can be made in many countries already using available forms of chemotherapy if treatment facilities are organized appropriately. PMID- 8242654 TI - Priorities for cancer control in Poland. AB - Deaths from cancer in Poland are expected to increase by over 60% during the next 20 years due to aging of the population and increasing age-standardized male cancer mortality. Five-year survivals are much lower than in developed Western countries. Improvement of statistics of cancer incidence and survival in different regions of Poland is essential for operational planning of interventions and resources. Reduction of tobacco smoking must remain a priority in prevention. Legislation enforcing protection from carcinogenic workplace exposures and development of monitoring is necessary. Poor performance of primary health care physicians is the major cause of delays in diagnosis. Interventions in this field should focus on professional training and development of units of rapid diagnosis of cancer. Implementation of state-of-the-art treatment through multicenter programs is recommended. Access to specialized treatment, in particular radiotherapy, should be improved through further development of the oncological networks. PMID- 8242655 TI - Cancer treatment in Poland: organization and policy. PMID- 8242656 TI - Partnership programs in Poland. AB - Sponsored by the U.S. Agency for International Development (AID), Albany Medical College and the Marie Sklodowska-Curie Memorial Cancer Center and Institute of Oncology formed a Partnership in 1992 aimed at enhancing cancer control and care in Poland by providing technical assistance and education to foster existing programs and develop new oncologic programs. The initial focus of the partnership is on breast cancer. Three demonstration screening/early disease programs are being developed, as well as a clinical and clinical research program and a Polish Breast Cancer Trials Group. A program to foster accreditation of oncologic facilities and programs throughout Poland is being considered. PMID- 8242657 TI - Prevention and screening for breast cancer. AB - Breast cancer is the most common cancer among women in both Europe and the U.S. It is the second leading cause of cancer deaths for women in the U.S., but the first in Poland. Age, family history, reproductive factors, proliferative breast disease, and radiation are established risk factors. Diet may account for differences in international incidence rates; however, the majority of women who get breast cancer do not have identifiable risk factors other than age and being female. Primary prevention of breast cancer is unknown. Control relies on secondary prevention aimed at suppression of clinically occult disease, mammographic screening, and early detection of palpable disease through physical examination and breast self-examination. Dietary and medical prevention strategies are being tested in the U.S. Screening mammography has demonstrated effectiveness in reducing mortality in women over 50. The "Partnerships in Healthcare" program aims to enhance the early detection of breast cancer in Poland by providing mammography units, training for technicians and nurses, and incentives to develop multidisciplinary approaches to diagnosis and treatment. PMID- 8242658 TI - Loco-regional management of early cancer of the breast. AB - Early detection of breast cancer and its treatment with breast conservation therapy (BCT) are major components of a strategy aimed at improving therapeutic outcome by treating disease earlier in its course. In this paper, we review the indications and contraindications for BCT, discuss principles of integrating the required diagnostic and therapeutic modalities, and present several areas of current controversy that pose the opportunity for appropriate clinical trials. PMID- 8242660 TI - Sequential changes in MHC antigen expression induced by the v-Ki-ras oncogene. AB - A series of early-passage cell lines were transformed with the v-Ki-ras oncogene with the aim of examining the effect of an activated ras gene on the ability of these cells to express major histocompatibility complex (MHC) antigens. These cell lines were found to undergo multiple phenotypic changes upon transformation and subsequent proliferation. At early passage, the predominant effect of ras was an increased ability to express class II antigens when induced with interferon gamma (IFN gamma). For class I antigens, maximum levels of expression induced with IFN gamma were largely unaffected, however, decreased sensitivity to induction with this lymphokine was noted. With subsequent in vitro or in vivo passage, both class I and class II antigen inducibility was attenuated. The latter phenotypic change was found to be transferable by coculture, implicating a soluble IFN gamma antagonist. Conditioned media from ras-transformed cells treated to activate their latent transforming growth factor beta (TGF beta) content mediated similar changes in MHC antigen inducibility, suggesting that TGF beta may be involved in modulating MHC antigen expression in ras-transformed cells. PMID- 8242661 TI - Phenotypic and functional profile of peripheral blood mononuclear cells isolated from melanoma patients undergoing combined immunotherapy and chemotherapy. AB - In the present study we tested the phenotypic profile as well as several immunological responses of peripheral blood mononuclear cells (PBMC) isolated from melanoma patients. These patients underwent chemotherapy with dacarbazine and carboplatin from day 1 to day 22, followed by immunotherapy of low-dose recombinant interleukin-2 and recombinant interferon alpha administered subcutaneously from day 36 to day 75. The PBMC from 14 patients were isolated on day 0 before chemotherapy, on day 36 after chemotherapy and on day 76 after immunotherapy. After chemotherapy, a decrease in CD16+ cells and increase in CD3+ and CD4+ cells correlated with a significant decrease in the generation of lymphokine-activated killer (LAK) activity. After immunotherapy, an increase in CD16+ cells correlated with an increase in the induction of LAK activity. A comparison between responding and non-responding patients revealed statistically significant differences in LAK activity of PBMC and response to concanavalin A following chemotherapy, and in the percentage of CD8+ cells following immunotherapy. Our results point toward the value of continuing such a study on a larger population of cancer patients in order to select the appropriate bioassays for monitoring and predicting the clinical responsiveness to combined therapies. PMID- 8242659 TI - Immune reconstitution following bone marrow transplantation. PMID- 8242662 TI - Tumor idiotype vaccines. VIII. Analysis of protective idiotype in sera and hybridomas derived from tumor-bearing mice with long-term survival. AB - In this study, the contribution of idiotype-positive antitumor antibodies (anti Id) in protective tumor immunity was investigated. We have previously shown that among various anti-Id generated and typed as the internal-image Ab2 of the tumor associated antibody (TAA) gp52, only 2F10 antibody induces protective immunity. Increase of the 2F10 idiotope in sera of tumor-bearing mice correlated with long term survival, while in mice with short survival the circulating 2F10 idiotype decreased. 2F10+ Ig were purified from sera of tumor-bearing mice with long-term survival and the amount of 2F10+ anti-TAA antibodies was determined. Only about 3% of 2F10+ antibodies are 2F10+ anti-TAA+. Hybridomas were generated from a 2F10 high mouse with spontaneous tumor regression. Only 2 out of 52 tumor-specific hybridomas were 2F10+. These results suggest that the protective effect induced by 2F10 vaccination may not be directly mediated by 2F10+ antibodies but indirectly through the stimulation of a 2F10-specific cellular immune response. PMID- 8242663 TI - Interleukin-4 plus tumor necrosis factor alpha augments the antigenicity of melanoma cells. AB - Immune cytokines are important regulators of the immune response to neoplastic cells. We previously reported that interleukin 4 (IL-4) and either tumor necrosis factor alpha (TNF) or interferon gamma (IFN) synergistically inhibit melanoma cell growth and induce cell differentiation. In the present study we used various combinations of IL-4, IFN and TNF to enhance the antigenicity of melanoma cells. IL-4 plus TNF significantly increased the ability of melanoma cells to stimulate cytotoxic T cells (CTL) and act as targets of these CTL; IL-4 plus IFN was somewhat less effective, while TNF plus IFN was not as effective. IL-4 plus TNF also increased the expression of HLA class I and HLA-DR antigens on melanoma cells. The CTL lines examined in this study were CD3+CD4+ and oligoclonal. These preclinical results suggest that the immune response to melanoma whole-cell vaccines might be enhanced by pretreating vaccine cells with IL-4 plus TNF. PMID- 8242664 TI - Increase in the ability of human cancer cells to induce cytotoxic T lymphocytes by ultraviolet irradiation. AB - The roles of ultraviolet-B (UV) radiation in the immunogenicity of human cancer cells have not been fully studied. We have investigated the effects of UV radiation on metastatic melanoma and renal cell carcinoma cells with regard to MHC antigen expression and the ability to induce cytotoxic T lymphocyte (CTL) activity in peripheral blood mononuclear cells (PBMC) or tumor-infiltrating lymphocytes (TIL) against untreated autologous tumor cells. UV radiation respectively decreased or increased MHC class I expression of freshly isolated tumor cells or cultured tumor cells, and also decreased MHC class I expression of starved cultured tumor cells. It increased the ability of both freshly isolated and cultured tumor cells to induce CTL activity from PBMC against untreated autologous tumor cells. UV-irradiated subclones that were more susceptible to CTL lysis were more potent for CTL induction from TIL than either an untreated parental clone or a UV-irradiated subclone that was resistant to CTL lysis. In summary, UV radiation increased the ability of tumor cells to induce CTL activity without a corresponding effect on MHC antigen expression. PMID- 8242666 TI - Turning the promises of computers into reality: Part 2. PMID- 8242665 TI - Effect of Adriamycin on liposomal muramyl tripeptide's ability to up-regulate monocyte cytokine expression. AB - Liposomal muramyl tripeptide phosphatidylethanolamine (L-MTP-PE) is a biological agent in phase I and II trials for osteosarcoma and melanoma. Its mechanism of action has been linked to its ability to activate monocyte tumoricidal function and to stimulate monocyte production of tumor necrosis factor (TNF) and interleukins(IL)-1, -6, and -8. Our ultimate goal is to combine L-MTP-PE with chemotherapy. The purpose of this study was to determine whether doxorubicin (Adriamycin) interfered with the ability of L-MTP-PE to activate monocyte cytokine production. Human monocytes were cultured with or without 5-500 ng/ml of Adriamycin for 3 h and washed before being exposed to 2 micrograms/ml L-MTP-PE for 16 h. Cultured supernatants were collected and assayed for TNF, IL-1, IL-6, and IL-8. The messenger RNA expression of IL-1 alpha, IL-1 beta, TNF alpha, IL-6, and IL-8 was quantified with northern blot analysis. Adriamycin did not suppress the up-regulation of any of these cytokines. We concluded that combination therapy with L-MTP-PE and Adriamycin is feasible and that this combination warrants further investigation in a clinical setting. PMID- 8242667 TI - Treatment of terminal dentitions: epidemiologic and clinical aspects of overdenture application. PMID- 8242668 TI - Angina pectoris-diagnosis and treatment in the outpatient setting. PMID- 8242669 TI - Class I and class II light-hardened glass-ionomer/resin restorations. PMID- 8242670 TI - Diagnostic procedures in oral amyloidosis. PMID- 8242671 TI - Affordable video imaging--still video photography. PMID- 8242672 TI - Malocclusion secondary to acromegaly: a case report. PMID- 8242673 TI - The hydroxyapatite ion activity product in acid solutions equilibrated with human enamel at 37 degrees C. AB - Five samples of powered enamel were each sequentially equilibrated 5 times at 37 degrees C with either 4 or 17 mmol/l H3PO4, in contact with air, and the ionic activity product for hydroxyapatite (IHA) estimated. There was evidence for preferential release of Mg and Na, especially in the first equilibrations. In two experiments, raised values of IHA were observed in the first equilibration but otherwise IHA was reasonable constant within experimental error and was much closer to the solubility product of hydroxyapatite than many previous estimates, mainly at 25 degrees C, suggest. The mean value of IHA for the majority fraction, averaged over all samples, was 1.7 (+/- 0.7) x 10(-58). Non-apatitic solids formed in all systems but solubility appeared nevertheless to be controlled by an apatitic phase: either the enamel mineral itself or apatite reprecipitated during the course of equilibration. High values of IHA reported previously may be due to use of conditions favouring dissolution only of more soluble factions or to metastability artifacts associated with control of solubility by non-apatitic phases. PMID- 8242674 TI - Effect of salivary components on dissolution rates of carbonated apatites. AB - Salivary pellicle has been reported to retard the acid-induced demineralization of dental enamel. However, this effect has not been quantified, nor has the direct protective effect of pellicle on carbonated apatite dissolution been elucidated. We have developed a model capable of further studying these questions. Well-characterized synthetic ceramic carbonated apatite discs were pretreated with parotid or whole salivary secretions for 18 h or 7 days and then challenged with 0.1 mol/l acetate, pH 5.0, in a dissolution apparatus. Surface dissolution rates were compared to those from the same carbonated apatites with no saliva pretreatment. Pellicles formed from whole and parotid saliva in vitro and whole saliva in vivo were capable of reducing surface dissolution rates of ceramic carbonated apatites by up to 83% but the degree of this reduction was dependent on the type of saliva and the conditions of pretreatment. PMID- 8242675 TI - Defluoridation of drinking water by co-precipitation with apatite. AB - The two-step brushite saturation, apatite precipitation method which has previously been shown to be effective for the partial defluoridation of water containing 10 ppm F has now been tested at lower concentrations. The method also reduced F concentrations in the range 1-5 ppm, but was less efficient at lower F levels. Although co-precipitation as fluorapatite is the intended mechanism here, experiments indicated that F adsorption on the seed material added to the defluoridation effect. When the contribution of co-precipitation was isolated it was found that this mechanism could, on its own, account for a reduction of 2.2 2.6 ppm F in water initially containing 3-5 ppm F. A computer program which determines equilibrium F concentrations when supersaturated solutions are induced to precipitate fluorapatite was used to estimate the maximum possible F removal from water by co-precipitation. Using saturation with brushite as a starting point for precipitation, F reductions similar to those found experimentally were predicted. Computer simulations also suggest that any desired reduction in F concentration may be achieved by adding sufficient Ca and phosphate and/or by raising the initial pH. PMID- 8242676 TI - The influence of different factors on in vitro enamel erosion. AB - The aim of this study was to use two demineralization test methods to analyze the erosive potential of beverages and foodstuffs. In addition, the surface microhardness test and the iodide permeability test were compared. Surface microhardness and iodide permeability were measured before and after exposure. To characterize the beverages and foodstuffs the content of phosphate, calcium and fluoride, pH, the titrable amount of base to pH 5.5 and 7.0 as well as the buffer capacity at pH 5.5 were determined. Sprite light showed the highest significant decrease in surface microhardness (p < or = 0.05) followed by grapefruit juice, apple juice and salad dressing. The highest significant increase in iodide permeability (p < or = 0.05) was caused by exposure to grapefruit juice followed by apple juice (Sprite light was not tested). Multiple linear regression analyses revealed that the erosive capacity of different drinks, juices and foodstuffs are statistically significantly associated with their acidity, pH values, phosphate and fluoride contents as well as the baseline surface microhardness or iodide permeability values of the exposed enamel. PMID- 8242677 TI - pH measurements of human dental plaque after consumption of starchy foods using the microtouch and the sampling method. AB - The aim was to study the effect of different starch-containing food items on pH changes in dental plaque. The pH was, in 10 subjects, simultaneously measured at various occasions up to 60 min in: (1) two approximal spaces in the premolar molar region with the microtouch method, and (2) pooled samples of dental plaque with the sampling method. Six starchy products were tested: (1) spaghetti, (2) macaroni, (3) potato, (4) rice, (5) unsweetened bread, and (6) sweetened bread. A glucose chewing tablet and a mouthrinse with 10% sucrose served as controls. Saliva was collected for HPLC analysis of maltose and maltotriose when testing the spaghetti, rice and unsweetened bread. The glucose tablet and the sucrose solution resulted in the greatest pH drops. However, all starchy products resulted in obvious pH falls, which were most pronounced for sweetened bread, followed, in order, by unsweetened bread and potato. Significantly more maltose and maltotriose were found in saliva after consumption of unsweetened bread compared to spaghetti. There seemed to be a relationship between the amount of starch hydrolysates in saliva and the area of the plaque pH curve (AUC). The microtouch method gave consistently lower pH values than the sampling method. Both methods, however, ranked the eight test products roughly in the same order with respect to minimum pH, maximum pH decrease and AUC. PMID- 8242678 TI - Microbiological validation of assessments of caries activity during cavity preparation. AB - The operative management of primary and secondary caries assumes that all discoloured tissue at the enamel-dentine junction (EDJ) represents active disease and this is removed to arrest the carious process. This study aims to establish clinical criteria to differentiate between active and arrested caries at the EDJ using microbiological assessment of dentine samples to verify its clinical status. Radiographs were available for posterior teeth. Cavities (n = 205) were prepared under rubber dam. After gaining access, areas of the EDJ were chosen and assessments made of consistency (soft, medium, hard), colour (dark brown, mid brown, pale) and moisture content (wet, dry). Dentine was removed by using a No. 3 round burr and placed in 1 ml of bacteriological culture broth. This sampling procedure was repeated at the same site once during cavity preparation and again when the cavity was judged as fully prepared. Samples were vortexed, diluted and cultured to give viable counts of the total anaerobic microflora, mutans streptococci and lactobacilli; viable counts were expressed as log10 (CFU per sample +1). Results showed no significant differences between the microflora of primary and secondary caries. The number of bacteria recovered diminished significantly as cavities were completed. Initial samples from soft and wet lesions harboured significantly more bacteria, lactobacilli and mutans streptococci than samples from medium, hard or dry lesions. Lesions visible on radiographs harboured more bacteria, including lactobacilli and mutans streptococci, while dentine colour was not discriminatory.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242679 TI - Comparison of different methods for the diagnosis of fissure caries without cavitation. AB - The aim of this study was to compare the accuracy of several common methods for the diagnosis of fissure caries. 63 human teeth without fillings and without any macroscopic carious cavitation but with different degrees of fissure discoloration and decalcification were selected from a pool. Dentists were asked to examine embedded teeth for fissure caries. The examination was done under standard conditions in a professional dental unit. The methods employed were: visual inspection (VI, n = 26 dentists), visual inspection with a magnifying glass (2x; VIM; n = 26), conventional bite-wing radiography (BW; n = 24), visual inspection combined with conventional bite-wing radiography (VI + BW; n = 10), and visual inspection combined with light pressure probing (VI + P; n = 23 dentists). In order to measure the reproducibility, VI and VI + BW were repeated. After the last inspection, the teeth were histologically prepared, serially sectioned perpendicular to the occlusal surface, and diagnosed for the presence of caries. The agreement between histological and clinical diagnosis was assessed. Specificities and sensitivities were: VI = 93 and 12%, VIM = 89 and 20%, BW = 83 and 45%, VI + BW = 87 and 49%, and VI + P = 93 and 14%, respectively. The following likelihood ratios were found: VI = 1.84, VIM = 1.86, BW = 2.6, VI + BW = 3.85, and VI + P = 2.05. It was concluded that the rather low sensitivity especially for VI and VI + P may lead to a significant number of teeth with dentinal caries being undetected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242680 TI - The effect of glass-ionomer cement on carious dentine: an in vivo study. AB - The effect of a glass-ionomer cement sealant on carious dentine was investigated clinically and bacteriologically. The study group consisted of 24 molar teeth, with small clinical visible occlusal dentine lesions, in 13 children (aged 7-18 years). Twenty of these molars were filled and sealed with a glass-ionomer cement (Fuji Ionomer Type III) and 4 with a resin sealant (Delton). From each molar, two dentine samples were collected aseptically with a time interval of 7 months. The first sample was taken after opening the lesion just beneath the dentino-enamel junction before application of the filling material (sample A), and the second beneath the removed filling material (sample B) 7 months later. Before collecting sample B the sealant was clinically evaluated and impressions were prepared in order to (re)evaluate the sealants later by scanning electron microscopy (SEM). After taking the second sample, the remaining fissures were opened to a standard depth, the caries which was still present was removed, and a composite resin restoration was applied. In all teeth, more glass-ionomer cement material was found by SEM compared to the clinical evaluation. In both the glass-ionomer- and the resin sealant-treated group, the median value of the total number of microorganisms (CFU) on blood agar was 100 times smaller in the B sample. Microorganisms were found in 90% of the B samples in the group treated with glass ionomer cement; hard dentine was also found in 45% of the B samples from this group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242681 TI - Prediction of dental caries in pre-school children. AB - Data obtained in a longitudinal study of caries incidence and caries-related factors were analysed with a view to producing a model for the prediction of caries. In direct correlations, caries incidence was significantly associated with bacterial, dietary and salivary variables; but when the data were examined by stepwise regression the strongest variables were the baseline caries score and misuse of sugar. Counts of Streptococcus mutans entered into the analysis but only as a relatively minor component. Similar significant relationships were seen with the determination of odds ratios. A caries activity test was formulated combining the following caries-associated variables: high counts of S. mutans, or lactobacilli, or the misuse of sugar, or frequent consumption of paediatric medicines. Regular use of fluoride tablets could compensate for paediatric medicine use or misuse of sugar. Such a caries activity test if it had been applied to the children at baseline would have had a positive predictive value of 0.76, a negative predictive value of 0.82, a sensitivity of 0.8 and a specificity of 0.78. Combining tests made the prediction of caries more accurate and in the population for which it was intended gave a reliable means of detecting those children most in need of enhanced caries prevention. PMID- 8242682 TI - Dental caries and cariogenic factors in pre-school urban Icelandic children. AB - Caries prevalence and several caries-related factors were measured in 158 pre school children in Reykjavik, initially aged 4 years and followed longitudinally for 2 years. The mean dmfs at 4 years was 3.3 and rose to 5.8 at 6 years although 34% remained caries-free throughout the study. Caries was very unevenly distributed. Children with dmfs > or = 5 at 4 years were significantly more likely to have caries in their permanent dentition at 6 years than those with a lower caries score at 4 years. Almost all children carried Streptococcus mutans but only 58% carried lactobacilli at 4 years. Lactobacilli declined in numbers at 5 and 6 years as the number of open carious lesions decreased but the mean count of S. mutans remained fairly static (> 10(5) cfu/ml). There were strong associations between high counts of S. mutans or lactobacilli and caries. Salivary parameters were not as strongly associated with caries although a low salivary pH and low salivary flow rate were significantly associated with high caries scores. The misuse of sugar as determined by dietary questionnaire was strongly associated with a high caries score and low caries prevalence was found in those children that took fluoride tablets regularly. PMID- 8242683 TI - Effect of erupting third molars on dental caries and gingival health in Finnish students. AB - The purpose of this study was to assess how the eruptive status of third molars relates to gingival and dental health and, in particular, to the status of the adjacent second molar. One hundred and twenty-three young adults were followed up over a 6-year period, from age 20 to age 26. The status of the second and third molars, and DMF, DMFS, DS, MS, and FS indices, periodontal condition, and salivary acidogenic bacterial counts were assessed. The 6-year DFS increase in relation to second molars was greatest in sextants with unerupted third molars. The percentage of erupting third molars that became carious or had been filled during the study period was 30%. Most new caries lesions or filled surfaces were on mesial surfaces of second molars and occlusal surfaces of third molars. The periodontal condition was best in mandibular sextants in which the third molar was missing. It is concluded that the most favourable situation for upper second molars is if the neighbouring third molar is missing by age 20. PMID- 8242684 TI - Effect of pinealectomy and the light/dark cycle on 2-[125I]iodomelatonin binding in the chick optic tectum. AB - 1. These studies investigated the regulation of melatonin receptor sites within the chick brain by the light/dark cycle and by endogenous melatonin using quantitative autoradiography. Saturation analyses performed in coronal brain sections of the optic tectum revealed a single class of high-affinity binding sites. 2. For diurnal rhythm studies, chicks were maintained on a 14:10 L:D cycle (light on 0400) and sacrificed at 4-h intervals. No significant variation was observed in either the affinity (KD = 67.8-76.4 pM) or the density (BMAX = 88.9 118.9 fmol/mg protein) of 2-[125I]iodomelatonin binding sites as a function of time of day. 3. In pinealectomized or age-matched sham-operated chicks, no changes were found in either the affinity (KD = 66.7-89.1 pM) or the density (BMAX = 112.8-180.4 fmol/mg protein) of 2-[125I]iodomelatonin binding sites at either 1, 7, or 14 days following surgery. These data suggest that endogenous pineal melatonin may not play a role in the regulation of melatonin receptor site affinity or density in the chick optic tectum. PMID- 8242685 TI - Down-regulation of beta-adrenergic and dopaminergic receptors induced by 2 phenylethylamine. AB - 1. The effects of chronic administration (28 days s.c. via Alzet osmotic minipumps) of 2-phenylethylamine.HCl (10 mg kg-1 per day) and/or (-)-deprenyl.HCl (1 mg kg-1 per day) on dopamine and noradrenaline receptor subtypes have been measured in rat brain. 3H-CGP 12177 was used to label beta-adrenoceptors; 3H spiperone and 3H-SCH 23390 were used to label D2-like and D1-like receptors. 2. Total cortical beta-adrenoceptor density was reduced by (-)-deprenyl but not 2 phenylethylamine alone. Combined administration of 2-phenylethylamine and (-) deprenyl resulted in a significantly larger decrease than (-)-deprenyl alone. Subtype density analysis by competition experiments with ICI 89406 revealed that the (-)-deprenyl effect in cortex was due to a decrease in beta 1-adrenoceptor density. The combination of 2-phenylethylamine and (-)-deprenyl resulted in a significant decrease in both cortical beta 1- and cortical beta 2-adrenoceptors. Cerebellar beta-adrenoceptor density was not altered by the present drug treatments. The Kd values for total beta-adrenoceptor densities and Ki values for beta-adrenoceptor subtype densities were not altered by drug treatment in either cortex or cerebellum. 3. Administration of 2-phenylethylamine and of (-)-deprenyl resulted in a decrease in the density of D1-like 3H-SCH 23390 but not D2-like 3H spiperone binding to dopamine receptors in the striatum. The effects of combined 2-phenylethylamine and (-)-deprenyl treatment on 3H-SCH 23390 binding were additive. These drug treatments did not alter Kd values for these binding sites. 4. The down-regulation of catecholamine receptors following chronically increased availability of 2-phenylethylamine may be due to the catecholamine releasing or uptake blocking effects of this amine. These effects may also be attributable to a direct neuromodulatory action of 2-phenylethylamine on catecholamine receptors. 5. The parallels between effects of increased 2-phenylethylamine availability and effects of administration of MAO inhibitor antidepressants on catecholamine receptor systems indicate that this substrate for MAO may mediate some of the effects of MAO inhibitor antidepressants. PMID- 8242686 TI - Calcium influxes and calmodulin modulate the expression and physicochemical properties of acetylcholinesterase molecular forms during development in vivo. AB - 1. Acetylcholinesterase (AcChoE; EC 3.1.1.7) exists in several molecular forms that may be anchored to cell membranes or associated with extracellular matrix. AcChoE bound to lipidic membranes is detergent extractable (DE AcChoE), whereas the enzyme associated with extracellular matrix is high salt soluble (HSS AcChoE). The latter variant is accumulated in synaptic regions by an unknown mechanism. 2. We have suggested previously that depolarization-induced Ca2+ influx is a major factor that modulates AcChoE synthesis in vivo, as well as the conversion of some DE AcChoE to HSS variant. In the present study, we have examined (i) the effects of depolarization-induced skeletal muscle inactivity and ionophore-induced Ca2+ influxes on the expression of AcChoE molecular forms and (ii) the hypothesis that Ca(2+)-dependent calmodulin may be involved in the conversion of at least some forms of DE AcChoE to HSS variant in vivo. 3. Chick embryos were treated in ovo during the early period of nerve-muscle interactions with d-tubocurarine (dTC; a competitive neuromuscular blocking agent) or with decamethonium (dMET; a depolarizing agent). Both dTC and dMET equally and significantly reduced embryonic neuromuscular activity (motility). However, dTC significantly decreased AcChoE overall activity, whereas dMET had virtually no effect on AcChoE expression, compared to controls. 4. Treatment of embryos with the Ca2+ ionophore A23187 significantly increased the total AcChoE activity as well as the DE/HSS ratio of each AcChoE molecular form. However, treatment with N (6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide (also termed W-7), a calmodulin antagonist, did not alter the total AcChoE activity, but significantly increased the DE/HSS ratio of AcChoE forms. 5. These results support the idea that (i) depolarization and/or Ca2+ influxes, but not muscle contraction, may regulate AcChoE expression in skeletal muscle and (ii) Ca(2+)-dependent calmodulin activation may be involved in the conversion of some DE AcChoE to their HSS variant in vivo. PMID- 8242687 TI - Quantitative receptor autoradiographic analysis for angiotensin II receptors in bovine retinal microvessels: quantitation with radioluminography. AB - 1. Specific 125I-Sar1, Ile8-Angiotensin II (125I-Sar1, Ile8-AII) binding sites in bovine retinal microvessels were investigated using the quantitative receptor autoradiographic method with pellet sections. 2. A quantitation was made with the computerized radioluminographic imaging plate system, a newly developed and highly sensitive method. Binding characteristics of the retinal microvessels were compared with those of the cerebral microvessels and the retinal macrovessels. 3. We isolated microvessels from the bovine retina and bovine cerebral cortex using the method composed of two-size sievings and high-speed homogenization with a Polytron. The isolated microvessels were composed of capillaries, and the retinal macrovessels contained vessels with smooth muscle. 4. There were specific binding sites for 125I-Sar1, Ile8-AII which were single and of a high affinity, in both the cerebral and the retinal microvessels and the retinal macrovessels. There were no differences in affinity between the vessels, but the retinal microvessels did have a higher density of binding sites than the cerebral microvessels. 5. The method we used is simple and sensitive for detecting and characterizing 125I Sar1, Ile8-AII binding sites in retinal capillaries. Knowledge of the existence of large numbers of specific binding sites, candidates of physiologically active angiotensin II receptors, aids with understanding the regulatory roles of angiotensin II in the blood-retinal barrier. PMID- 8242688 TI - Chronic caffeine alters the density of adenosine, adrenergic, cholinergic, GABA, and serotonin receptors and calcium channels in mouse brain. AB - 1. Chronic ingestion of caffeine by male NIH strain mice alters the density of a variety of central receptors. 2. The density of cortical A1 adenosine receptors is increased by 20%, while the density of striatal A2A adenosine receptors is unaltered. 3. The densities of cortical beta 1 and cerebellar beta 2 adrenergic receptors are reduced by ca. 25%, while the densities of cortical alpha 1 and alpha 2 adrenergic receptors are not significantly altered. Densities of striatal D1 and D2 dopaminergic receptors are unaltered. The densities of cortical 5 HT1 and 5 HT2 serotonergic receptors are increased by 26-30%. Densities of cortical muscarinic and nicotinic receptors are increased by 40-50%. The density of cortical benzodiazepine-binding sites associated with GABAA receptors is increased by 65%, and the affinity appears slightly decreased. The density of cortical MK-801 sites associated with NMDA-glutaminergic receptors appear unaltered. 4. The density of cortical nitrendipine-binding sites associated with calcium channels is increased by 18%. 5. The results indicate that chronic ingestion of caffeine equivalent to about 100 mg/kg/day in mice causes a wide range of biochemical alterations in the central nervous system. PMID- 8242689 TI - The isolation of a cDNA encoding a neuropeptide prohormone from the light yellow cells of Lymnaea stagnalis. AB - 1. The central nervous system (CNS) of the freshwater snail Lymnaea stagnalis contains several clusters of neuroendocrine cells, which synthesize neuropeptides that act as neurotransmitters, neurohormones, and/or neuromodulators, controlling a broad range of physiological processes. Using a protein chemical approach, we have previously characterized a peptide [named LYCP-A (Hoek et al., 1992], which is produced by the neuroendocrine light yellow cells (LYC), which are present as two clusters of endogenously bursting neurons in the visceral and right parietal ganglion, respectively. 2. A differential screening technique was used to isolate the cDNA that encodes the prohormone of LYCP-A. The prohormone appeared to contain three or four putative neuropeptides, one of which is LYCP-A. The organization of the identified prohormone resembles that of the histidine-rich basic peptide precursor previously identified in the R3-14 neurons of the marine snail Aplysia californica (Campanelli and Scheller, 1987). 3. In situ hybridization analysis indicates that the gene encoding the LYC prohormone is expressed in a subset of the LYC. The LYC release their peptides into the hemolymph from a neurohemal area, which is located around the CNS. In addition, the peptides are released from axonal branches in the aorta of the heart, suggesting a role in the regulation of cardiovascular functions. PMID- 8242690 TI - Identification of a 7B2-derived tridecapeptide from bovine adrenal medulla chromaffin vesicles. AB - 1. A novel tridecapeptide was isolated from extracts of bovine adrenal medulla chromaffin vesicles and the primary structure determined to be SVPHFSDEDKDPE. 2. This peptide is identical to the C termini of human and porcine 7B2 and is highly homologous to the same region of the mouse and Xenopus lavis protein. 3. In all these species the homologous peptide is preceded by a pair of lysine residues, a potential proteolytic processing site. 4. Ser6 is part of a well-conserved casein kinase II consensus phosphorylation sequence. Evidence for phosphorylation of this residue was obtained during Edman sequencing. 5. Thus, this novel adrenal medullary probably arises from the posttranslational processing of the bovine 7B2 protein. PMID- 8242691 TI - Proteolysis at the secretase and amyloidogenic cleavage sites of the beta-amyloid precursor protein by acetylcholinesterase and butyrylcholinesterase using model peptide substrates. AB - 1. It was recently proposed that acetylcholinesterase (AChE), in addition to its esteratic activity, has proteolytic activity such that it may cleave the beta amyloid precursor (beta-APP) within the beta-amyloid sequence. The purpose of this paper was to examine further whether AChE or butyrylcholinesterase (BuChE) had associated proteinase activity that was involved in the metabolism of beta APP. 2. The ability of various preparations of AChE and BuChE to hydrolyze two synthetic fragments of beta-APP695 as model substrates containing the normal and aberrant cleavage sites was studied. 3. Digestion of these synthetic substrates with commercial preparations of Electrophorus electricus AChE indicated the presence of a trypsin-like proteolytic activity cleaving each peptide at the carboxy-terminal side of an internal lysine residue. 4. Purification of the trypsin-like proteinase activity by aminobenzamidine affinity chromatography yielded a preparation that was devoid of AChE activity but retained all of the proteinase activity. 5. Amino-terminal sequence analysis of this preparation showed that the first 13 amino acid residues were identical to beta-pancreatic trypsin. 6. These data indicate that the proteinase activity found in these commercial preparations of AChE is due to contamination with trypsin. PMID- 8242692 TI - [Congenital heart defects in adults worldwide and in the Czech Republic]. AB - Congenital heart disease of adults is a new area of cardiology with specific diagnostic, therapeutic, social and economic problems of these patients. The First Medical Clinic of the Second Medical Faculty, Charles University assembled so far data on more than 1200 adult patients with congenital heart disease from different districts in Bohemia and Moravia. Atrial septal defects are recorded most frequently (25.7%), followed by ventricular septal defects (25.0%) and coarctation of the aorta (12.4%). 50.3% of the patients were treated by surgery, most frequently patients with a patent ductus arteriosus--in 83.8%, with coarctation of the aorta (82.8%) and an atrial septal defect (60.5%). The authors review reasons why for the care of these patients in other countries specialized centres developed and the foundation of such a centre in the Czech Republic is discussed. PMID- 8242693 TI - [Pericardial puncture under echocardiographic control]. AB - The authors summarize their hitherto assembled experience with pericardiocentesis under echocardiographic control. From May 1986 to December 1991 they made these punctures 55 times in 49 patients. In the majority therapeutic puncture was involved, only seven times puncture was used for diagnostic purposes. Echocardiography can evaluate not only the presence of an exudate and assess its haemodynamic impact but it is the optimal method for finding an optimal site for puncture and it is very suitable for checking the course of the operation. In smaller operations and in biopsies of pathological pericardial tissue it is advisable to use special instruments which make the procedure safe. The authors recorded a total of three failures, in one instance the exudate was removed with the assistance of a surgeon, in one instance the situation was coped with by repeated puncture and in one instance by a conservative procedure. None of the patients developed complications. A list of the latter is discussed. The method is nowadays part of therapeutic procedures of the cardiologist. PMID- 8242694 TI - [Significance of hetero-collateral circulation in patients with chronic occlusion of the superficial femoral artery]. AB - The authors examined a total of 11 lower extremities in 10 patients with chronic occlusion of the superficial femoral artery (mean length of occlusion 17.2 cm) in order to compare the participation of the heterocollateral and homocollateral circulation in the maintenance of the ankle pressures assessed by the Doppler principle. Before percutaneous transluminal recanalization of the chronic occlusion of the superficial femoral artery the authors induced by means of a balloon catheter short-term occlusion of the deep femoral artery in its central portion, closely after its insertion (complete block of the heterocollateral circulation) and in the periphery of this artery (partial block of the collateral circulation). During both types of balloon occlusion the authors assessed the ankle pressures and compared them with control values, assessed before occlusion. The authors found that in their patients the heterocollateral circulation, as compared with the homocollateral one, participated substantially more in the value of the ankle pressure at rest. The importance of the heterocollateral circulation increased with the length of chronic occlusion of the superficial femoral artery. In some patients the homocollateral circulation did not participate at all in the maintenance of the ankle pressure. A drop of the ankle pressure in patients with chronic occlusion of the superficial femoral artery is also more marked in acute occlusion of the deep femoral artery in its central portion, as compared with acute occlusion in the periphery. PMID- 8242695 TI - [The oral magnesium loading test for detecting possible magnesium deficiency]. AB - In 26 top sportsmen (volleyball players, rower) aged 15-18 years the serum magnesium level and urinary Mg excretion before and after oral administration of 5 g Mg lactate was assessed (oral magnesium load test). In 11 sportsmen (42.3%) a low percentage of excretion of administered Mg was found, suggesting magnesium deficiency, irrespective of the basal serum Mg levels. After subsequent 10-day Mg supplementation 6 volleyball players responded by a markedly lower retention after an identical Mg load, again without marked changes of the serum Mg level. The authors consider the magnesium load test a more sensitive indicator of latent Mg deficiency than the serum Mg level which is maintained at a relatively stable level and declines only in severe deficiency. The test assumes normal renal function and intestinal absorption. PMID- 8242696 TI - [Possible therapeutic applications of pulsed magnetic fields]. AB - Magnetotherapy is a relatively new, nowadays however, relatively widespread method in several medical disciplines. The mechanism proper of the favourable action of the pulsed magnetic field on the living organism is not quite clear so far, clinical investigations revealed, however, a favourable anti-inflammatory, angioedematous and analgesic therapeutic effect. The authors sought an optimal frequency of the pulsed magnetic field with regard to the character of the disease. They focused attention above all on treatment of acute and chronic inflammatory conditions of the locomotor apparatus, ischaemia of the blood vessels of the lower extremities, dyspeptic syndrome, lactation mastitis and other diseases. One therapeutic cycle lasted 20 minutes, the mean number of cycles varied between 5.8 and 7.7. A regression of complaints was recorded as a rule after 2-3 sessions. The optimal frequency of the pulsed magnetic field seems to be a value between 10.0 and 25.0 Hz. It is useful in particular in severe conditions to repeat the therapeutic cycle after 2-3 months. The advantage of this therapeutic method is the minimal number of contraindications. PMID- 8242697 TI - [Thoughts on ethical problems of long-term institutional care of chronically ill patients and the aged]. AB - Long-term institutional care of dependent subjects is an integral part of chronic health and social care. Its importance will increase as a result of the increasing numbers of chronic patients and subjects depending on the care of others. The author reviews some serious problems associated with care provided in institutes of social care, pensioners' homes and long term care departments in our country and abroad. She discusses the cultural and historical aspect of care, the contemporary state and quality of care and investigates the causes of negative attitudes to these institutions and possibilities how to improve care and the quality of life of their inmates. PMID- 8242698 TI - [Interesting items from the personal effects of Jan Evangelista Purkinje]. PMID- 8242699 TI - [Treatment and prevention of addictions in the USA]. PMID- 8242700 TI - Carbamylcholine-induced morphological changes and spatial dynamics of [Ca2+]c in Harderian glands of guinea pigs: calcium-dependent lipid secretion and contraction of myoepithelial cells. AB - To determine whether lipid-secreting cells have cytosolic Ca2+ concentration ([Ca2+]c)-related secretory mechanisms, morphological changes and intracellular calcium dynamics of Harderian glands of guinea pigs stimulated by secretagogue were studied by electron microscopy and Fura-2/AM digital image analysis. Control glandular cells contained large lipid vacuoles that were bordered by multi layered membranes. Rough-surfaced endoplasmic reticulum, mitochondria, and smooth surfaced endoplasmic reticulum may be involved in lipid vacuole formation. Myoepithelial cells surrounded alveoli. After carbamylcholine (CCh, 10(-6), 10( 5), and 10(-3) M) stimulation, lipid materials within the membranous structures were frequently discharged by an exocytotic mechanism. Conspicuous deformation of glandular cells caused by vigorous contraction of myoepithelial cells was observed in isolated alveoli after 10(-6) M CCh stimulation, whereas the deformities of glandular tissues perfused via vessels were small even after 10( 3) M CCh stimulation. Connective tissue between glandular alveoli inhibited unbridled myoepithelial-cell contraction. Fura-2/AM digital imaging analysis revealed that CCh stimulation caused an increase in [Ca2+]c in isolated alveoli. The morphological reactions and changes in [Ca2+]c were prevented by atropine. When extracellular calcium ions were absent, enhanced extrusion of lipid vacuoles, myoepithelial-cell contraction, and a rise in [Ca2+]c after CCh stimulation were not observed. Nicotine and catecholamines had no effect on the secretion or on the dynamics of [Ca2+]c. It can be concluded that acetylcholine elicits exocytosis in glandular cells and contraction of the myoepithelial cells of Harderian glands, accompanied by an increase in [Ca2+]c. The dynamics of [Ca2+]c of the gland alveoli are mostly dependent on extracellular Ca2+. PMID- 8242701 TI - Localization of spectrin isoforms in the adult mouse heart. AB - The distribution of two isoforms of spectrin in the adult mouse heart was investigated by Western blotting and immunocytochemistry by use of monospecific antibodies to erythrocyte spectrin and nonerythroid brain spectrin (240/235). Western blotting revealed proteins analogous to both isoforms of alpha-spectrin in adult heart. Light-microscopic immunocytochemistry indicated that erythroid spectrin was distributed throughout the myocardium, with immunofluorescence localized to plasma membranes, Z-lines, and intercalated discs. Antibodies to brain spectrin (240/235) exhibited staining throughout the heart, with a generally diffuse distribution except for the prominent immunoreactivity associated with the intercalated discs. Nonerythroid spectrin immunofluorescence was detected in the endothelial cells of the endocardium and the mesothelial cell lining of the epicardium. Erythrocyte spectrin was not detected in the endocardium or the epicardium. The identification and localization of spectrin isoforms in the mammalian heart suggest the importance of spectrin proteins in the structural integrity and proper function of cardiac cells and tissues. This is the first demonstration of two different alpha-spectrin subunits in the mammalian heart. PMID- 8242702 TI - 20-OH-ecdysone swells nuclear volume by alkalinization in salivary glands of Drosophila melanogaster. AB - Ecdysteroids play an important role in the larval moulting process of insects. Ecdysone-induced stimulation causes specific "puffs" in polytene chromosomes of salivary gland cells resulting in nuclear swelling. During this process, changes of intracellular ion composition are thought to act as an early regulatory mechanism of gene activation. By use of video-imaging analysis and electrophysiological techniques, we examined ecdysone-induced nuclear swelling in Drosophila salivary glands in situ and its dependence on pH and calcium. Isolated glands of the third larval stage were superfused with a solution mimicking the haemolymph. Addition of 5 x 10(-6) mol/1 20-OH-ecdysone led, after a lag period of 50 min, to a sustained Ca(2+)-dependent increase of nuclear volume by 23.0 +/- 2.3%. Amiloride, a blocker of plasma membrane Na+/H+ exchange, prevented 20-OH ecdysone-induced nuclear swelling. Decreasing pH in the superfusate from 7.15 to 6.8 led to nuclear shrinkage by 16.9 +/- 3.9%. Measurements of pH in salivary gland cells with ion-sensitive microelectrodes disclosed an alkalinization of 0.23 +/- 0.05 pH units after stimulation with 20-OH-ecdysone. We postulate that 20-OH-ecdysone activates the amiloride-sensitive plasma membrane Na+/H+ exchanger. This leads to intracellular alkalinization and concomitant decondensation of the nuclear chromatin visible as nuclear swelling. Thus, cell alkalinization could be a potentially important stimulatory mechanism in mediating ecdysteroid-induced activation of the cell nucleus. PMID- 8242703 TI - Distribution of immunoreactive Tamm-Horsfall protein in various species in the vertebrate classes. AB - A sheep antibody to human Tamm-Horsfall protein, the major protein in normal urine, was used in an immunohistological study of organs of 48 species of vertebrate animals, representing the classes Mammalia, Aves, Reptilia, Amphibia, Osteichthyes and Chondrichthyes. Immunoreactivity was shown in the thick limb of the loop of Henle in the kidney of mammals, but there was no reactivity with tissues of birds or reptiles. Superficial layers of the skin of several amphibians and fish, superficial layers of the oral mucosa and gills of fish, and the distal tubules of the kidney of some amphibians, reacted with the antibody. Immunoreactivity with mammalian kidney was removed by passage of the antibody down an immunoadsorption column coated with human Tamm-Horsfall protein, and amphibian immunoreactivity was removed by incubation of the antibody with material prepared from frogs in the same way as Tamm-Horsfall protein. These findings suggest that immunoreactive Tamm-Horsfall protein appeared early in vertebrate phylogeny, initially in skin and gills and later in kidney, and that although conserved in evolution, it shows antigenic differences between amphibians and mammals. Its distribution is consistent with the hypothesis that is acts as a waterproofing agent. PMID- 8242704 TI - Response of the seminiferous epithelium of the rat testis to withdrawal of androgen: evidence for direct effect upon intercellular spaces associated with Sertoli cell junctional complexes. AB - The morphological response of the Sertoli cells to partial or complete withdrawal of testosterone was studied in adult rats following hypophysectomy or administration of ethane dimethanesulphonate (EDS), a toxicant known to destroy selectively the Leydig cells of the testis. To assess the role of germ cells in effecting changes to Sertoli cells following withdrawal of testosterone, germ cell-deficient rats with Sertoli-cell-only testes (SCO) were treated with EDS to remove the source of testosterone. At 6 days after hypophysectomy or 4, 6 and 8 days after EDS treatment, stage VII and VIII seminiferous tubules showed degenerating germ cells and numerous basally-located vacuoles approximately 1-15 microns in diameter. Ultrastructural analysis indicated that most of the vacuoles were multiple focal dilations of the intercellular space associated with Sertoli cell junctional complexes. In SCO rats, treatment with EDS resulted in a significant (P < 0.05) increase in the formation of many vacuoles particularly in the base but also in the trunk of the Sertoli cells and again electron microscopic analysis showed multiple, localized expansions of the intercellular space associated with Sertoli cell junctional complexes. The appearance of intercellular spaces in SCO testes following androgen withdrawal cannot be attributed to shrinkage of degenerating germ cells since the seminiferous tubules did not contain germ cells. It is concluded that withdrawal of androgen induces early morphological alterations of the Sertoli cell junctional complexes in which the sites of membrane fusions representing tight junctions remain intact whereas the intercellular spaces exhibit major focal dilations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242705 TI - Development and androgen regulation of the secretory cell types of the Syrian hamster (Mesocricetus auratus) Harderian gland. AB - The secretory cell types of the hamster Harderian glands were studied in both male and female Syrian hamsters. As previously demonstrated, female hamsters showed a single secretory cell type (type I), while male hamsters displayed two secretory cell types (type I and type II). Type-II cells were observed after the first month of age correlating with the increase in testosterone levels. The administration of testosterone to adult female hamsters resulted in a marked increase in the percentage of type-II cells without a significant increase in the number of mitotic figures. Very low levels of serum testosterone were able to maintain the percentage of type-II cells. Castration of male hamsters produced a decrease in the percentage of type-II cells. This drop correlated with the reduction in serum testosterone levels. The chronic administration of a luteinizing hormone-releasing hormone agonist to male Syrian hamsters induced a significant reduction in both serum luteinizing hormone and testosterone. However, the percentage of type-II cells was similar to that of control hamsters suggesting that very low levels of circulating testosterone are able to maintain the percentage of type-II cells. In a final experiment male Syrian hamsters were treated with the antiandrogen cyproterone acetate. No changes were observed in the percentage of type-II cells, whereas serum luteinizing hormone and testosterone levels were significantly modified. We concluded that (1) type-II cells differentiate from type-I cells; (2) gonadal androgens are the major factor controlling this differentiation; and (3) the disappearance of type-II cells after androgen deprivation occurs through holocrine and apocrine mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242706 TI - Mouse adrenal chromaffin cells can transform to neuron-like cholinergic phenotypes after being grafted into the brain. AB - The development of neuron-like cholinergic immunophenotypes by adrenal chromaffin cells was studied in 10-week-old mouse adrenal medullary grafts. Fragments of chromaffin tissue were implanted into mouse hippocampus, and antibodies specific for neurofilaments (NF), neuron-specific enolase (NSE), choline acetyltransferase (ChAT), acetylcholinesterase (AChE), and phenylethanolamine-N-methyltransferase (PNMT) were applied to the grafts. Adrenal medulla grafts survived well and most of the transplanted cells were either round or polygonal. A minority of chromaffin cells elaborated an intermediate or sympathetic neuron phenotype. Chromaffin cells showed pronounced immunoreactivity for NSE in their perikarya and axon-like processes: immunoreactivity for NF was only found in a few processes. In adjacent immunohistochemically stained sections, the transplanted cells stained for ChAT and AChE. At the electron-microscope level, the immunohistochemical reactions for the two acetylcholine-related enzymes were mainly located on the endoplasmic reticulum and in cell processes. Immunoreactivity for PNMT was found to decline in transplanted chromaffin cells below that of normal adrenal medulla. These observations suggest that, in adrenal medullary grafts implanted into the hippocampus, chromaffin cells are endowed with neuron-like cholinergic immunophenotypes. PMID- 8242707 TI - Demonstration of melatonin-binding sites in cyclohexylamine-formaldehyde-fixed brain tissues. AB - This study shows for the first time that perfusion of rat or hamster brain with a cyclohexylamine-paraformaldehyde mixture makes possible the observation by autoradiography of melatonin binding sites in structurally well-preserved fixed tissues. This result is a first step in the identification of melatonin-receptor containing cell types by cytoautoradiography. PMID- 8242708 TI - Immunocytochemical approach to the structure of human blood group B and H glycoconjugate antigens in the three days old rat cochlea. AB - The possible structure of human blood-group antigens, as found in cochlear hair cells of 3-day-old rats, is suggested. Data were obtained from immunocytochemical studies using 77 antibodies against the major human blood group antigens of the ABO, H, I and Lewis genetic systems. Neither the anti-A-related nor the anti Lewis-related antibodies showed any positive immunoreaction on hair cells. In contrast, anti-B, anti-AB and anti-H antibodies displayed specific positive immunoreactive patterns on the hair cells. The results suggest that, in immature hair cells, two main glycoconjugate structures of the lactoseries are present: H type 2 antigen, which is the precursor of the B type 2 antigen, and the B type 2 antigen itself. Similar H and B carbohydrate structures have been reported in rat olfactory receptors. The type 2 glycoconjugates carrying these H and B antigens of auditive and olfactory receptors are resistant to fixation and paraffin embedding, suggesting that they might be glycoproteins. These auditive and olfactory H and B antigens must be different from the B-related antigens that are expressed by pseudo-unipolar neurons of rat posterior root ganglia, that are built from type 4 core chains, and that are destroyed by routine paraffin embedding procedures. PMID- 8242709 TI - Insect myotropic peptides: differential distribution of locustatachykinin- and leucokinin-like immunoreactive neurons in the locust brain. AB - Locustatachykinin I is one of four closely related myotropic neuropeptides isolated from brain and corpora-cardiaca complexes of the locust Locusta migratoria. Antiserum was raised against locustatachykinin I for use in immunocytochemistry. It was found that the antiserum recognizes also locustatachykinin II and hence probably also the other two locustatachykinins due to their similarities in primary structure. Locustatachykinin-like immunoreactive (LomTK-LI) neurons were mapped in the brain of the locust, L. migratoria. A total of approximately 800 LomTK-LI neurons were found with cell bodies distributed in the proto-, deuto- and tritocerebrum, in the optic lobes and in the frontal ganglion. Processes of these neurons innervate most of the synaptic neuropils of the brain and optic lobes, as well as the frontal ganglion and hypocerebral ganglion. The widespread distribution of LomTK-LI neurons in the locust brain indicates an important role of the locustatachykinins in signal transfer or regulation thereof. As a comparison neurons were mapped with an antiserum against the cockroach myotropic peptide leucokinin I. This antiserum, which probably recognizes the native peptide locustakinin, labels a population of about 140 neurons distinct from the LomTK-LI neurons (no colocalized immunoreactivity). These neurons have cell bodies that are distributed in the proto- and tritocerebrum and in the optic lobe. The processes of the leucokinin-like immunoreactive (LK-LI) neurons do not invade as large areas in neuropil as the LomTK-LI neurons do and some neuropils, e.g. the mushroom bodies, totally lack innervation by LK-LI fibers. In some regions, however, the processes of the LomTK LI and LK-LI neurons are superimposed: most notably in the central body and optic lobes. A functional relation between the two types of neuropeptide in the locust brain can, however, not be inferred from the present findings. PMID- 8242710 TI - Immunocytochemical localization of oviduct-specific glycoproteins in the oviductal epithelium from cows at follicular and luteal phases. AB - The immunocytochemical localization of bovine oviduct-specific glycoproteins was investigated by light and electron microscopy. Using monoclonal antibodies (MAbs) specific for bovine oviductal glycoproteins, 3 regions (fimbriae, ampulla, and isthmus) of the epithelium in the bovine oviduct we studied during the follicular and luteal phases. The MAbs reacted specifically with the oviductal epithelial cells. Intense labeling was observed in the ampullar and fimbrial epithelia of cows at the follicular phase, but the reactions were weaker at the luteal phase. In the isthmus, the immunohistochemical reaction was faint during both follicular and luteal phases. At the ultrastructural level, the MAbs bound selectively to putative secretory granules of nonciliated cells in the ampulla and fimbriae, but not in the isthmus. These results suggest that there are cyclic changes and regional differences in the production of glycoproteins in the bovine oviduct. PMID- 8242711 TI - Localization of corazonin in the nervous system of the cockroach Periplaneta americana. AB - Antisera raised to the cardioactive peptide corazonin were used to localize immunoreactive cells in the nervous system of the American cockroach. Sera obtained after the seventh booster injection were sufficiently specific to be used for immunocytology. They recognized a subset of 10 lateral neurosecretory cells in the protocerebrum that project to, and arborize and terminate in the ipsilateral corpus cardiacum. They also reacted with bilateral neurons in each of the thoracic and abdominal neuromeres, a single dorsal unpaired median neuron in the suboesophageal ganglion, an interneuron in each optic lobe, and other neurons at the base of the optic lobe, in the tritocerebrum and deutocerebrum. The presence of corazonin in the abdominal neurons and the lateral neurosecretory cells was confirmed by HPLC fractionation of extracts of the abdominal ganglia, brains and retrocerebral complexes, followed by determination of corazonin by ELISA, which revealed in each tissue a single immunoreactive peak co-eluting with corazonin in two different HPLC systems. Antisera obtained after the first three booster injections recognized a large number of neuroendocrine cells and neurons in the brain and the abdominal nerve cord. However, the sera from the two rabbits reacted largely with different cells, indicating that the majority of this immunoreactivity was due to cross-reactivity. These results indicate that the production of highly specific antisera to some neuropeptides may require a considerable number of booster injections. PMID- 8242712 TI - The shape and distribution of lysosomes and endocytosis in the ciliary epithelial cells of rats. AB - The shape and distribution of lysosomes in the ciliary epithelium of rat eyes were examined by electron microscopy combined with acid phosphatase (ACPase) cytochemistry and three-dimensional observation of 2 microns-thick sections. ACPase activity was cytochemically localized in lysosomes and trans Golgi cisternae in the non-pigmented epithelial (NPE) and pigmented epithelial (PE) cells. In NPE cells, it was shown three-dimensionally, that most lysosomes had an elongate form, up to 5 microns in length, and a diameter of 70-100 nm. These elongate lysosomes (nematolysosomes) were predominantly located in the basal region of the cells. In contrast, PE cells had spherical lysosomes distributed at random throughout the cytoplasm. However, no nematolysosomes were seen in the PE cells. When the isolated ciliary processes were incubated in a medium containing horseradish peroxidase (HRP), HRP was incorporated into the nematolysosome-like structures by pinocytosis from the basal surface of the NPE cells. These findings suggest that nematolysosomes are associated with the pinocytotic activity of NPE cells. The pinocytosis-nematolysosomal route may be involved in the uptake and degradation of macromolecules from the aqueous humor in the posterior chamber. PMID- 8242713 TI - Rod-opsin immunoreaction in the pineal organ of the pigmented mouse does not indicate the presence of a functional photopigment. AB - The aim of the present study was to characterize the rod-opsin immunoreaction in the mammalian pineal organ. Pigmented mice (strain C57BL) were selected as the animal model. Immunocytochemical investigations involving the use of highly specific polyclonal and monoclonal antibodies against bovine rod-opsin (the apoprotein of the photopigment rhodopsin) showed that approximately 25% of all pinealocytes were rod-opsin immunoreactive. Immunoblotting techniques revealed three protein bands of approximately 40, 75, and 110 kDa; these were detected by the monoclonal antibody and the polyclonal antiserum in retinal and pineal extracts. These protein bands presumably represented the monomeric, dimeric and trimeric forms of rod-opsin. The amount of rod-opsin in retina and pineal organ was quantified by means of an enzyme-linked immunosorbent assay. This yielded 570 +/- 30 pmoles rod-opsin per eye and 0.3 +/- 0.05 pmoles rod-opsin per pineal organ. High pressure liquid chromatography analysis of whole eye extracts demonstrated the chromophoric group of the photopigment rhodopsin, 11-cis retinal, and its isomer, all-trans-retinal. A shift from 11-cis retinal to all trans-retinal was found upon light adaptation. No retinals were detected in the pineal organ. Autoradiographic investigations showed that 3H-retinol, intraperitoneally injected into the animals, was incorporated into the outer and inner segments of retinal photoreceptors, but not into the pineal organ. It is concluded that the mouse pineal organ contains the authentic apoprotein of rhodopsin but that it lacks retinal derivatives as essential components of all known vertebrate photopigments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242714 TI - Vasoactive intestinal peptide-immunoreactive cerebrospinal fluid-contacting neurons in the reptilian lateral septum/nucleus accumbens. AB - By means of immunocytochemical demonstration of vasoactive intestinal peptide (VIP) an accumulation of cerebrospinal fluid (CSF)-contacting neurons was found in a circumscribed region of the nucleus accumbens/lateral septum of eleven reptilian (chelonian, lacertilian, ophidian, crocodilian) species. Basal processes of these cells contribute to a subependymal plexus whose density displays considerable interspecific variation. VIP-immunoreactive nerve fibers occur also in the lateral septum and the nucleus accumbens where they encompass immunonegative cells in a basket-like pattern. The CSF-contacting neurons are surrounded by columnar ependymocytes frequently arranged in a pseudostratified manner. These specialized arrays of ependymal cells, however, occupy a more extended area than the VIP-immunoreactive CSF-contacting neurons and can be traced from the rostro-ventral pole of the lateral ventricle to the interventricular foramen. These observations suggest the existence of a telencephalic site of CSF-contacting neurons which may be more widespread than hitherto thought and which may participate in a circumventricular system of the lateral ventricle. Previous studies mainly performed with birds indicate that the VIP-immunoreactive CSF-contacting neurons of the nucleus accumbens might form a part of the "encephalic" (extraretinal and extrapineal) photoreceptor. However, further experiments are required to test this supposition since the VIP immunoreactive neurons of the nucleus accumbens remained unlabeled by antibodies against bovine rodopsin and chicken cone-opsin in all eleven species analysed in this investigation. PMID- 8242715 TI - Cell shape and arrangement of cultured aortic smooth muscle cells grown on collagen gels. AB - Aortic smooth muscle cells (SMC) grown on conventional plastic culture dishes have morphological and functional properties of dedifferentiated cells in subculture. We examined the influence of collagen gels on the cell shape and arrangement. The cells grown on collagen gels showed a multilayered growth with formation of nodules. When the edge of the collagen gels was detached from the culture dish, the shape and arrangement of cells on the edge differed from that of the central, still attached region. The cells grown on floating collagen gels exhibited a spindle-like shape and were arranged in concentric circles. These findings suggest that the physical property of the substrate influences the cell shape and arrangement. PMID- 8242716 TI - Modes of neuronal arbor enlargement in the ear of a postembryonic fish, Astronotus ocellatus. AB - New hair cells are added during postembryonic life in several species of fishes and birds. The production of new hair cells appears to require enlargement of eighth nerve arbors during growth since, at least in fish, eighth nerve neurons are added more slowly than hair cells or not at all. This situation provides an intriguing opportunity to study the mechanisms of growth of the neuronal arbors. In this paper, we report the results of studies on the postembryonic growth of eighth nerve dendritic arbors in the saccular epithelium of the cichlid fish Astronotus ocellatus. Arbor sizes and shapes were compared in small and large fish using the axonal tracer cobaltous lysine. Our data suggest that postembryonic eighth nerve arbors enlarge in 2 ways. First, arbors add new terminal endings to their distal ends. Second, whole new branches appear to be added at locations up to hundreds of micrometers proximal to the terminal endings. These 2 modes of growth suggest that more than one mechanism may be operative in controlling arbor enlargement. PMID- 8242717 TI - Enhancement of cell killing and increase in cytosolic calcium concentration by combined treatments with hyperthermia and TMB-8 in mouse mammary carcinoma FM3A cells. AB - Mouse mammary carcinoma FM3A cells were exposed 42 degrees C or 44 degrees C hyperthermia for 60 min in a culture medium with or without 8-(N,N-diethylamino) octyl 3,4,5-trimethoxybenzoate hydrochloride (TMB-8) at a concentration of 100 microM. Cell survival, cytosolic calcium ion concentration, [Ca2+]C, and influx of 45Ca2+ were investigated to examine the cardinal role of intracellular Ca2+ in hyperthermic cell killing. The [Ca2+]C in a single cell was measured by a digital image analyzing technique using Fura-2. TMB-8 enhanced cell killing, increase in [Ca2+]C, and influx of 45Ca2+ induced by both hyperthermic treatments. In contrast, TMB-8 abolished the increase in [Ca2+]C induced by hyperthermia at 42 degrees C and 44 degrees C in the absence of extracellular Ca2+. These results suggest that TMB-8, an agent usually called an inhibitor of intracellular calcium mobilization at 37 degrees C or lower, enhances Ca2+ entry from the extracellular space at 42 degrees C or 44 degrees C and, in turn, promotes cell killing under hyperthermic conditions. PMID- 8242718 TI - Control of intracellular Ca2+ by adrenergic and muscarinic agonists in mouse mandibular ducts and end-pieces. AB - The changes in free Ca2+ ([Ca2+]i) in the cells of the secretory end-pieces and intralobular ducts of mouse mandibular glands exposed to adrenergic or cholinergic agonists were measured using fluorescence imaging techniques. [Ca2+]i in both cell types increased in a dose-dependent manner during both adrenergic and cholinergic stimulation. The duct cells responded to noradrenaline and to acetylcholine over the same concentration range (30 nmol/l to 3 mumol/l) although the maximum increase in [Ca2+]i above resting levels evoked by noradrenaline (ca. 137 nmol/l) was about twice that evoked by acetylcholine. The response to acetylcholine was blocked by atropine (0.1 mumol/l) and the response to noradrenaline was blocked by the alpha 1-adrenergic antagonist, prazosin (0.1 mumol/l), but not by the alpha 2-adrenergic antagonist, yohimbine. The alpha adrenergic agonist, phenylephrine, mimicked the action of noradrenaline but the beta-adrenergic agonist, isoproterenol, had no effect. In contrast to the duct cells, the end-piece cells responded to acetylcholine at much lower concentrations (threshold << 1 nmol/l) than to noradrenaline (threshold ca. 300 nmol/l) and the size of the increase in [Ca2+]i above resting levels evoked by acetylcholine (216 nmol/l) was nearly 5-times greater than for noradrenaline. VIP and substance P failed to evoked a Ca2+ response in either end-piece or duct cells. PMID- 8242719 TI - Characterization of a polyclonal antiserum against the purified human recombinant calcium binding protein calretinin. AB - We have purified recombinant human calretinin (CR) from Escherichia coli lysates and have produced a polyclonal antiserum against it. The antiserum recognizes determinants conserved in fish, chicken, rat, monkey and human CR. We show its use in the qualitative detection of CR by different methods of immunohistochemistry as well as in the detection of CR on immunoblots. PMID- 8242720 TI - Differential activation of inositol 1,4,5-trisphosphate-sensitive calcium pools by muscarinic receptors in Xenopus laevis oocytes. AB - Muscarinic acetylcholine (ACh) receptors activate the phospholipase C signal transduction pathway to promote the formation of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and the consequent elevation of cytoplasmic calcium (Ca2+). The inositol phosphate and Ca(2+)-mobilization responses to ACh were analyzed in Xenopus oocytes possessing endogenous receptors, and in oocytes expressing exogenous receptors from injected muscarinic RNA transcripts, to evaluate the patterns of signal transduction mediated by native and expressed receptors. Activation of native ACh receptors elicited dose- and time-dependent increases in Ins(1,4,5)P3 and inositol bisphosphate (InsP2) production. ACh-induced Ins(1,4,5)P3 production increased rapidly within the first 2 min and continued to rise over the next 20 min. ACh was a much more effective stimulus of inositol phosphate production at native (up to 35-fold) than at expressed receptors (less than 2-fold). In contrast, measurements of Ca(2+)-mobilization in oocytes injected with the Ca(2+)-specific photoprotein, aequorin, revealed that ACh stimulation of expressed receptors evoked up to 200-fold increase in light emission, whereas ACh stimulation of native receptors elicited less than a 2-fold response. These observations indicate that the oocyte possesses functionally distinct agonist-sensitive Ca2+ pools which differ markedly in their sensitivity to Ins(1,4,5)P3 production and suggest that these pools are mobilized by different effector mechanisms. The finding that the magnitude of the intra-oocyte Ca2+ response is not necessarily determined by the degree of Ins(1,4,5)P3 production, but rather by another aspect of the signal transduction pathway (e.g. the nature and/or location of the Ins(1,4,5)P3 releasable Ca2+ pool), reveals an additional level of complexity in the transduction mechanisms responsible for intracellular Ca2+ signaling. PMID- 8242721 TI - Dopamine receptor imaging. PMID- 8242722 TI - A review of hormonal findings in cluster headache. Evidence for hypothalamic involvement. AB - The cause of cluster headache remains to be determined. The involvement of peripheral neurovascular structures can explain the pain and autonomic signs of a cluster attack, but not its rhythmicity. The central theory of cluster headache attributes the cyclic recurrence to involvement of the hypothalamus. To evaluate hypothalamic dysfunction a number of hormone studies have been carried out on cluster headache patients. Alterations in plasma melatonin, cortisol, testosterone, gonadotrophins, prolactin, growth hormone and thyrotropin have been documented, some only in the cluster period but others in the remission phase of the illness. We believe that the hormonal abnormalities in cluster headache support disorders of hypothalamic function. PMID- 8242723 TI - Prevalence of headache in Tulp's Observationes Medicae (1641) with a description of cluster headache. AB - In 1641, Nicolaas Tulp, a well-known physician from Amsterdam, published his Observationes Medicae, in which medical knowledge and practice in that period is described in more than 200 systematically organized case histories. Next to symptomatic headaches he mentioned two different types of "recurring headache" based on the characteristics of the pain and reaction to contemporary treatment: migraine and, probably, cluster headache. PMID- 8242724 TI - Painful stimulation of the forehead increases photophobia in migraine sufferers. AB - Thresholds for visual and auditory discomfort were investigated in 51 migraine sufferers and 27 controls of similar age and sex distribution who rarely suffered from headache. Tests in migraine sufferers were carried out during the headache free interval. Discomfort thresholds were measured before and during painful stimulation of the forehead with ice. The visual discomfort threshold was lower in migraine sufferers than in controls, and decreased further during painful stimulation of the forehead. In contrast, the auditory discomfort threshold was similar in migraine sufferers and controls, and did not decrease during painful stimulation of the forehead. These findings suggest that trigeminal discharge contributes to photophobia but not phonophobia in migraine sufferers. PMID- 8242725 TI - Dopamine D2-receptor imaging with 123I-iodobenzamide SPECT in migraine patients abusing ergotamine: does ergotamine cross the blood brain barrier? AB - Two migraine patients were studied by in vivo SPECT using the dopamine D2 receptor specific radioligand 123I-3-iodo-6-methoxybenzamide (123I-IBZM) during ergotamine abuse and after withdrawal. Results were compared with 15 healthy controls. Striatum/cerebellum and striatum/occipital cortex ratios of count rate density were calculated as a semiquantitative measurement for striatal dopamine D2-receptor binding potential. No differences were found in striatal uptake of 123I-IBZM between healthy controls and the patients when on or off ergotamine. Preliminary evidence suggests that ergotamine may not occupy striatal dopamine D2 receptors to a large extent and thus may not cross the blood brain barrier in large quantities. PMID- 8242726 TI - Prevalence of frequent headache in young Finnish adults starting a family. AB - A questionnaire study on pre-pregnancy headache and its predictors was carried out in a representative sample of young adults expecting their first baby. The study was part of a major project on the competence of Finnish families. It included 1322 women and 1257 men, either married or cohabiting (in total, 1262 families), followed from their first contact with a maternity health care unit. A questionnaire was given separately to the women and men inquiring about their health and health behaviour, as well as about sociodemographic and psychosocial factors. The prevalence of frequent headache (at least one episode per month) was 34.3% in women, 19.3% in men and 47.1% in families. On multivariate analysis, after age-adjustment, the independent predictors of headache occurrence in women were depression, menstrual pain, responsibilities at work and psychosocial stress. The predictors in men were nervousness, psychosocial stress, lack of or irregular physical exercise and long-term disease, depression, responsibilities at work and insomnia. PMID- 8242727 TI - The trigemino-pupillary response in cluster headache. AB - Central impairment of the integrative neural systems controlling vegetative function and pain perception has been demonstrated in cluster headache (CH). Recently, we described the human pupillary response (trigeminal reflex) to quantified (painless and painful) corneal stimulation with a combined neurophysiological and pharmacological technique. In this study, the trigeminal reflex was evaluated in 26 subjects with episodic cluster headache. During the active phase of the disease, on the side of the pain we observed reduced mydriasis to electrical stimuli with an intensity equal to the corneal reflex threshold, and on both sides to stimuli with intensity that equalled the pain threshold. No difference was found when amplitude of the miotic phase was compared in the different groups. These suggest disordered pupillary activation in response to pain, probably sympathetic in origin, which is bilateral, detectable also during the remission phase and which cannot be explained simply by the antidromic release of pain-related peptides. PMID- 8242728 TI - Platelet-rich plasma serotonin levels in tension-type headache and depression. AB - We measured platelet-rich plasma (PRP) serotonin in patients suffering from tension-type headache, before and after treatment with amitriptyline, comparing them with a healthy control group and patients with untreated depression. We evaluated the severity of headache and depression in each group. PRP serotonin levels were higher in patients with headache than in controls and depressed patients. We observed a fall of PRP serotonin in patients with tension-type headache to similar levels after treatment as the depressed group. This fall was correlated with the improvement of headache but not with depression scales. Our data suggest that the rise of platelet serotonin levels in tension-type headache is related to pain and not depression. PMID- 8242729 TI - Increased platelet serotonin content and hypersecretion from dense and alpha granules in vitro in tension-type headache. AB - We investigated platelet aggregation and secretion from dense and alpha-granules in vitro in 28 tension-type headache (TH) patients and 26 healthy controls. We also measured basal platelet serotonin levels. Platelet aggregation was normal in TH, but the secretion of serotonin and platelet factor 4 (PF4) was significantly increased in response to 0.5 and 2.0 micrograms/ml collagen and to 1.0 mumol/l PAF. The basal platelet serotonin levels were also higher in patients than in controls. The mechanisms of platelet hypersecretion remain to be determined, but the increased secretion of serotonin is probably in part related to the increased basal levels. The increased platelet serotonin in TH patients may reflect an enhanced serotonin turnover. PMID- 8242730 TI - Occipital neuralgia treated by intradural dorsal nerve root sectioning. AB - The term occipital neuralgia was first used in 1821 to describe a characteristic pain in the region innervated by the greater occipital nerve. Since that time numerous authors have failed to reach a consensus concerning the presentation, cause, and treatment of this entity. Review of these previous works reveals that the term occipital neuralgia encompasses a collection of signs and symptoms which develop secondary to a variety of different disease processes. We have treated nine patients suffering from severe occipital pain with associated C2 hypesthesia, unilateral retro-orbital pain, and occipital or C2 tubercle trigger points by intradurally sectioning the C1-C4 dorsal rootlets (C4 if shoulder pain existed). All patients had been treated previously with various modalities without success. This paper discusses the results obtained in these patients. PMID- 8242731 TI - Trigeminal neuralgia with chronic paroxysmal hemicrania: the CPH-tic syndrome. AB - A case of chronic paroxysmal hemicrania-tic syndrome is described. The chronic paroxysmal hemicrania (CPH) was controlled with indomethacin and the trigeminal neuralgia with a glycerol blockade. The trigeminal neuralgia reappeared four years after the blockade and was then treated successfully with corticosteroids. Six months later, when indomethacin was stopped, the trigeminal neuralgia recurred and resolved again with corticosteroids. We suggest that the CHP-tic syndrome is due to periorbital venous vasculitis. To our knowledge this is the first reported case of this disorder. PMID- 8242732 TI - Problem drinkers do not get headache. PMID- 8242733 TI - Protein translocation across the endoplasmic reticulum: a tunnel with toll booths at entry and exit. PMID- 8242734 TI - Coatomers and SNAREs in promoting membrane traffic. PMID- 8242735 TI - Friends and family: the role of the Rab GTPases in vesicular traffic. PMID- 8242736 TI - Sorting of membrane proteins in the secretory pathway. PMID- 8242737 TI - Incorrect attribution of peptide source. PMID- 8242738 TI - Protein translocation into proteoliposomes reconstituted from purified components of the endoplasmic reticulum membrane. AB - We have reproduced the process of protein transport across and of protein integration into the mammalian endoplasmic reticulum membrane by the use of proteoliposomes reconstituted from pure phospholipids and purified membrane proteins. The transport of some proteins requires only two membrane protein complexes: the signal recognition particle receptor, needed for targeting of a nascent chain to the membrane, and a novel complex, the Sec61p complex, that consists of Sec61p and two smaller polypeptides. The translocation of other proteins also needs the presence of the translocating chain-association membrane (TRAM) protein. The integration of two membrane proteins of different topologies into the membrane does not require additional components. These results indicate a surprising simplicity of the basic translocation machinery. They suggest that the Sec61p complex binds the ribosome during translocation and forms the postulated protein-conducting channel. PMID- 8242739 TI - Genetic identification of Mom-1, a major modifier locus affecting Min-induced intestinal neoplasia in the mouse. AB - Mutations in the human APC gene caused various familial colon cancer syndromes. The Multiple intestinal neoplasia (Min) mouse provides an excellent model for familial colon cancer: it carries a mutant mouse Apc gene and develops many intestinal adenomas. Here, we analyze how this tumor phenotype is dramatically modified by genetic background. We report the genetic mapping of a locus that strongly modifies tumor number in Min/+ animals. This gene, Mom-1 (Modifier of Min-1), maps to distal chromosome 4 and controls about 50% of genetic variation in tumor number in two intraspecific backcrosses. The mapping is supported by a LOD score exceeding 14. Interestingly, Mom-1 lies in a region of synteny conservation with human chromosome 1p35-36, a region of frequent somatic loss of heterozygosity in a variety of human tumors, including colon tumors. These results provide evidence of a major modifier affecting expression of an inherited cancer syndrome. PMID- 8242740 TI - The C. elegans cell death gene ced-3 encodes a protein similar to mammalian interleukin-1 beta-converting enzyme. AB - We have cloned the C. elegans cell death gene ced-3. A ced-3 transcript is most abundant during embryogenesis, the stage during which most programmed cell deaths occur. The predicted CED-3 protein shows similarity to human and murine interleukin-1 beta-converting enzyme and to the product of the mouse nedd-2 gene, which is expressed in the embryonic brain. The sequences of 12 ced-3 mutations as well as the sequences of ced-3 genes from two related nematode species identify sites of potential functional importance. We propose that the CED-3 protein acts as a cysteine protease in the initiation of programmed cell death in C. elegans and that cysteine proteases also function in programmed cell death in mammals. PMID- 8242741 TI - Induction of apoptosis in fibroblasts by IL-1 beta-converting enzyme, a mammalian homolog of the C. elegans cell death gene ced-3. AB - The mammalian interleukin-1 beta-converting enzyme (ICE) has sequence similarity to the C. elegans cell death gene ced-3. We show here that overexpression of the murine ICE (mICE) gene or of the C. elegans ced-3 gene causes Rat-1 cells to undergo programmed cell death. Point mutations in a region homologous between mICE and CED-3 eliminate the ability of mICE and ced-3 to cause cell death. The cell death caused by mICE can be suppressed by overexpression of the crmA gene, a specific inhibitor of ICE, as well as by bcl-2, a mammalian oncogene that can act to prevent programmed cell death. Our results suggest that ICE may function during mammalian development to cause programmed cell death. PMID- 8242742 TI - Identification of human activin and TGF beta type I receptors that form heteromeric kinase complexes with type II receptors. AB - Transforming growth factor beta (TGF beta) and activin each bind to pairs of membrane proteins, known as receptor types I and II, that associate to form a signaling complex. We report that TSR-I and ActR-I, two human transmembrane serine/threonine kinases distantly related to TGF beta and activin type II receptors, act as type I receptors for these factors. TSR-I is a type I receptor shared by TGF beta and activin, whereas ActR-I is an activin type I receptor. ActR-I, but not TSR-I, signals a particular transcriptional response in concert with activin type II receptors. The results indicate that type I receptors are transmembrane protein kinases that associate with type II receptors to generate diverse heteromeric serine/threonine kinase complexes of different signaling capacities. PMID- 8242743 TI - Cloning of a TGF beta type I receptor that forms a heteromeric complex with the TGF beta type II receptor. AB - A cDNA clone encoding a 53 kd serine/threonine kinase receptor with an overall structure similar to that of the type II receptor for transforming growth factor beta (TGF beta) was obtained. 125I-TGF beta 1 bound to porcine endothelial cells transfected with the cDNA and formed a cross-linked complex of 70 kd, characteristic of a TGF beta type I receptor. Immunoprecipitation of the cross linked complexes by antibodies against the cloned receptor revealed the 70 kd complex as well as a 94 kd TGF beta type II receptor complex. The immunoprecipitated novel serine/threonine kinase receptor had biochemical properties of the TGF beta type I receptor and was observed in different cell types. Transfection of the cloned cDNA into TGF beta type I receptor-deficient cells restored TGF beta-induced plasminogen activator inhibitor 1 production. These results suggest that signal transduction by TGF beta involves the formation of a heteromeric complex of two different serine/threonine kinase receptors. PMID- 8242744 TI - Three-dimensional structure of recoverin, a calcium sensor in vision. AB - Recoverin, a recently discovered member of the EF hand superfamily, serves as a calcium sensor in vision. We report here the crystal structure of recombinant unmyristoylated recoverin at 1.9 A resolution. The four EF hands of the protein are arranged in a compact array that contrasts with the dumbbell shape of calmodulin and troponin C. A calcium ion is bound to EF hand 3, while EF hand 2 can bind samarium but not calcium in this crystal form. The other two EF hands have novel structural features that prevent or impair calcium binding. A concave hydrophobic surface formed by EF hands 1 and 2 may participate in the read out of calcium signals by recoverin and its homologs. PMID- 8242745 TI - Loss of a yeast telomere: arrest, recovery, and chromosome loss. AB - Yeast strains were constructed in which a single telomere could be eliminated from the end of a dispensable chromosome. In wild-type cells, elimination of a telomere caused a RAD9-mediated cell cycle arrest, indicating that telomeres help cells to distinguish intact chromosomes from damaged DNA. However, many cells recovered from the arrest without repairing the damaged chromosome, replicating and segregating it for as many as ten cell divisions prior to its eventual loss. Telomere elimination caused a dramatic increase in loss of the chromosome in all strains examined, demonstrating that yeast telomeres are also essential for maintaining chromosome stability. Thus, in spite of checkpoint and DNA damage repair systems, many chromosomes that lose a telomere are themselves destined for loss. PMID- 8242746 TI - Interaction between dorsal and ventral cells in the imaginal disc directs wing development in Drosophila. AB - The adult appendages of Drosophila develop from imaginal discs. An early step in disc patterning involves the formation of developmental boundaries that subdivide the discs into compartments. Anterior and posterior compartments are established in the embryo. Later in development a new boundary originates to subdivide the wing disc into dorsal and ventral compartments, which correspond to the dorsal and ventral surfaces of the adult wing. We report here that spatially localized expression of the homeobox gene apterous (ap) specifies the identity of dorsal cells in the wing. The boundary of cell lineage restriction between dorsal and ventral compartments coincides with the limit of the domain of ap expression. Using genetic mosaics, we show that juxtaposition of dorsal and ventral cells induces formation of the wing margin. We present evidence that the dorsal-ventral boundary promotes growth and serves as a pattern-organizing center in the wing disc. PMID- 8242747 TI - Dif, a dorsal-related gene that mediates an immune response in Drosophila. AB - There are striking parallels between the regulation of gene expression along the dorsoventral (DV) axis of Drosophila embryos and lymphoid-restricted expression in the mammalian immune system. Both depend on regulatory factors containing rel domains (dorsal and NF-kappa B) that are controlled at the level of nuclear transport. A novel Rel-containing gene in Drosophila, Dif (dorsal-related immunity factor), provides a potential link between these seemingly disparate processes. Although Dif maps close to dorsal, it does not appear to participate in DV patterning, but instead mediates an immune response in Drosophila larvae. Dif is normally localized in the cytoplasm of the larval fat body, but quickly accumulates in the nucleus upon bacterial infection or injury. Evidence is presented that once in the nucleus, Dif binds to kappa B-like sequence motifs present in promoter regions of immunity genes. These results suggest that mammalian and insect immunity share a common evolutionary origin. PMID- 8242748 TI - Differential induction of transcriptionally active p53 following UV or ionizing radiation: defects in chromosome instability syndromes? AB - Accumulation of p53 protein was seen in the nuclei of mammalian cells following DNA damage caused by ultraviolet radiation (UV), X-ray, or a restriction enzyme. Promoters containing p53-binding sites show a dramatic transcriptional response to DNA damage. The p53 response to X-ray is rapid, reaching a peak at 2 hr after radiation, but is very transitory and reduced in magnitude compared with that seen in response to UV. We find no substantive defect in the p53 response of cells from ataxia telangiectasia or xeroderma pigmentosum complementation group A patients. In contrast, 2 out of 11 primary cultures from Bloom's patients showed a complete absence of p53 accumulation following UV irradiation or SV40 infection and a grossly delayed and aberrant response following X-ray. PMID- 8242749 TI - A C-terminal protein-binding domain in the retinoblastoma protein regulates nuclear c-Abl tyrosine kinase in the cell cycle. AB - The ubiquitously expressed c-Abl tyrosine kinase is localized to the nucleus and binds to DNA. The DNA binding activity is regulated by cdc2-mediated phosphorylation, suggesting a cell cycle function for c-Abl. Here we show that the tyrosine kinase activity of nuclear c-Abl is regulated in the cell cycle through a specific interaction with the retinoblastoma protein (RB). A domain in the C-terminus of RB, outside of the A/B pocket, binds to the ATP-binding lobe of the c-Abl tyrosine kinase, resulting in kinase inhibition. The RB-c-Abl interaction is not affected by the viral oncoproteins that bind to RB. Hyperphosphorylation of RB correlates with release of c-Abl and activation of the tyrosine kinase in S phase cells. The nuclear c-Abl tyrosine kinase can enhance transcription, and this activity is inhibited by RB. Nuclear c-Abl is an S phase activated tyrosine kinase that may participate directly in the regulation of transcription. PMID- 8242750 TI - Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2. AB - We used the interaction trap, a yeast genetic selection for interacting proteins, to isolate human cyclin-dependent kinase interactor 1 (Cdi1). In yeast, Cdi1 interacts with cyclin-dependent kinases, including human Cdc2, Cdk2, and Cdk3, but not with Ckd4. In HeLa cells, Cdi1 is expressed at the G1 to S transition, and the protein forms stable complexes with Cdk2. Cdi1 bears weak sequence similarity to known tyrosine and dual specificity phosphatases. In vitro, Cdi1 removes phosphate from tyrosine residues in model substrates, but a mutant protein that bears a lesion in the putative active site cysteine does not. Overexpression of wild-type Cdi1 delays progression through the cell cycle in yeast and HeLa cells; delay is dependent on Cdi1 phosphatase activity. These experiments identify Cdi1 as a novel type of protein phosphatase that forms complexes with cyclin-dependent kinases. PMID- 8242751 TI - The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. AB - The cyclin-dependent kinase Cdk2 associates with cyclins A, D, and E and has been implicated in the control of the G1 to S phase transition in mammals. To identify potential Cdk2 regulators, we have employed an improved two-hybrid system to isolate human genes encoding Cdk-interacting proteins (Cips). CIP1 encodes a novel 21 kd protein that is found in cyclin A, cyclin D1, cyclin E, and Cdk2 immunoprecipitates. p21CIP1 is a potent, tight-binding inhibitor of Cdks and can inhibit the phosphorylation of Rb by cyclin A-Cdk2, cyclin E-Cdk2, cyclin D1 Cdk4, and cyclin D2-Cdk4 complexes. Cotransfection experiments indicate that CIP1 and SV40 T antigen function in a mutually antagonistic manner to control cell cycle progression. PMID- 8242752 TI - WAF1, a potential mediator of p53 tumor suppression. AB - The ability of p53 to activate transcription from specific sequences suggests that genes induced by p53 may mediate its biological role as a tumor suppressor. Using a subtractive hybridization approach, we identified a gene, named WAF1, whose induction was associated with wild-type but not mutant p53 gene expression in a human brain tumor cell line. The WAF1 gene was localized to chromosome 6p21.2, and its sequence, structure, and activation by p53 was conserved in rodents. Introduction of WAF1 cDNA suppressed the growth of human brain, lung, and colon tumor cells in culture. Using a yeast enhancer trap, a p53-binding site was identified 2.4 kb upstream of WAF1 coding sequences. The WAF1 promoter, including this p53-binding site, conferred p53-dependent inducibility upon a heterologous reporter gene. These studies define a gene whose expression is directly induced by p53 and that could be an important mediator of p53-dependent tumor growth suppression. PMID- 8242753 TI - A novel moesin-, ezrin-, radixin-like gene is a candidate for the neurofibromatosis 2 tumor suppressor. PMID- 8242754 TI - Synergistic effect of extracellular adenosine 5'-triphosphate and tumor necrosis factor on DNA degradation. AB - Extracellular ATP (ATPo) was recently considered a possible mediator of cell mediated cytotoxicity since it is secreted by effector cells following appropriate stimulus and causes lysis as well as DNA degradation of susceptible target cells. This hypothesis however is not readily reconciled with the finding that ATPo-resistant cells are fully susceptible to intact effector cells, which instead suggests that a necessary step in cell-mediated cytotoxicity is the interaction between different molecules released by a cytotoxic cell. By combining ATPo with TNF or lymphotoxin (LT), cytokines which induce late DNA damage, we observed a synergistic effect on target cell death. Under these conditions, the phenomenon of DNA degradation also appeared early, with a kinetics reminiscent of that observed during target cell incubation with intact effector cells. Target cells which are resistant to one of the two molecules exhibited an enhanced rate of cell death when exposed to their association. Target cell lysis and DNA degradation were also Ca(2+)-independent events as they took place following external Ca2+ chelation by EGTA addition and under experimental conditions in which little or no Ca2+ was released from target cell intracellular stores. These findings suggest that ATPo might represent a further mediator which is responsible for the alternative Ca(2+)-independent cytolytic pathway in association with other molecules released by effector cytotoxic lymphocytes. PMID- 8242755 TI - Correlation between induction of lymphocyte apoptosis and prostaglandin E2 production by macrophages infected with HIV. AB - Several data indicate that HIV infection of antigen-presenting cells (APCs) and apoptosis of lymphocytes play important roles in pathogenesis of AIDS. We have recently demonstrated that prostaglandin E2 (PGE2) can cause thymocyte apoptosis in vivo. In the present study we have investigated the possibility that the intercellular contacts between HIV-infected APCs and lymphocytes could induce apoptosis in the latter population and that PGE2-production by HIV-infected APCs could be involved in the hypothesized phenomenon. Monocytes/macrophages separated from peripheral blood mononuclear cells (PBM phi) of healthy donors were infected in vitro and maintained in culture. PGE2 was promptly produced by HIV-infected PBM phi and values of PGE2 concentrations in supernatants over the background noninfected controls persisted for more than 2 weeks. HIV-infected PBM phi or cell-free supernatants from their cultures were then added to autologous uninfected lymphocytes and apoptosis was assessed by morphological criteria and by looking to the expression of tissue transglutaminase, one of the effector elements of the program of cell death. In both culture conditions the percentage of apoptotic lymphocytes was significantly increased in respect to values obtained in control cultures. When a cyclooxygenase inhibitor was added to the HIV-infected PBM phi cultures, the percentage of apoptotic lymphocytes was reduced at levels similar to those observed after cocultivation with uninfected PBM phi or exposure to supernatants from uninfected PBM phi. In addition, a substantial increase in apoptosis in lymphocytes from healthy donors was found following PGE2 treatment in vitro. PMID- 8242756 TI - Selective prothymocyte targeting by prenatal diethylstilbesterol exposure. AB - Estrogens have been reported to modulate immunologic responses at both physiologic and pharmacologic concentrations. Treatment of experimental animals with the synthetic estrogen, diethylstilbesterol (DES), markedly decreases thymic cellularity, manifested histologically as a progressive loss of cortical thymic lymphocytes. In the present report thymic atrophy after prenatal DES exposure was found to be more severe than has been reported following adult exposure, indicating a possible greater sensitivity of the developing immune system to estrogenic hormones. DES exposure resulted in a limited alteration of cell maturation within the fetal thymus as evidenced by only slight alterations in the expression of CD4 and CD8 cell-surface antigens. To examine the possibility that DES targets hematopoietic stem cells in the fetal liver, cytometric analysis was conducted using a panel of fluorescent antibodies to quantitate the hematopoietic subpopulations present in control and DES-exposed Gestational Day (gd) 18 fetal mouse liver. There were no significant DES-induced alterations in the number of hematopoietic stem cells, or in fetal liver cells expressing CD44 (hematopoietic precursors), Mac-1 (granulocyte-macrophage lineage precursors), or CD45R (B lineage lymphocytes) surface antigens. However, DES selectively reduced the number of fetal liver precursors containing the lymphocyte stem cell-specific DNA polymerase, terminal deoxynucleotidyl transferase, which suggested that DES may specifically target the fetal liver prothymocyte. Reconstitution of irradiated hosts with gd 18 fetal liver from vehicle and DES-exposed syngeneic donors demonstrated an impaired ability of the DES-treated fetal liver to repopulate the thymus of irradiated hosts. In addition, fetal liver cells enriched for prelymphoid cells contained potentially significant levels of estrogen specific receptors. Taken together these data, in conjunction with the lack of direct thymocyte injury (necrosis, apoptosis, and/or inhibition of cell proliferation) by DES treatment, suggest that estrogen-mediated thymic atrophy may result, at least in part, from a specific alteration in the lymphocyte stem cell population responsible for colonizing the thymus. PMID- 8242757 TI - CD3 monoclonal antibodies evoke the same cytochrome P450-regulated capacitative entry of calcium as thapsigargin in Jurkat T cells. AB - In T cells CD3 monoclonal antibodies mediate an elevation of cytosolic Ca2+ concentration due to a release from internal stores and also due to an entry from extracellular medium, the mechanism of which is not clearly elucidated. Previous studies on several cell types have reported that depleting intracellular Ca2+ stores with inhibitors of the reticulum Ca(2+)-ATPase resulted in an increased plasma membrane permeability to calcium ions. It has been suggested that emptying the reticulum triggers a Ca2+ influx from extracellular medium, independent of phosphoinositide hydrolysis. To document the physiological relevance of such a mechanism, we compared CD3- and thapsigargin-induced sustained increase of cytosolic Ca2+ concentration in Jurkat T cells with regard to their sensitivity to internal and external Ca2+ level and to several inhibitors which do not affect the release of internal stores. We show that (1) there was no additivity of the two effects; (2) both CD3- and thapsigargin-evoked Ca2+ influx were inhibited when membrane was depolarized by either gramicidin or a high potassium concentration; and (3) Ca2+ influx was abrogated by cytochrome P450 inhibitors such as lipoxygenase inhibitors or imidazole antimicotic drugs. CD3 mAb and thapsigargin thus triggered the same signaling events, probably involving a cytochrome P450, to transmit information from depleted endoplasmic reticulum to the plasma membrane. PMID- 8242758 TI - The interaction of auranofin and buthionine sulfoximine blocks activation of human peripheral T lymphocytes. AB - We have previously demonstrated that auranofin, at nanomolar concentrations, enhances T cell activation, as measured by IL-2 release and Tac expression. However, it is not clear how enhanced T cell activation might be related to therapeutic value, since rheumatoid arthritis is widely believed to be associated with overactivation of the immune response. In this study, we show that the action of auranofin on T cell activation is dramatically influenced by the glutathione levels of the responding cells. Under conditions of very low intracellular glutathione where the synthesis of glutathione is blocked, the action of auranofin is converted from enhancement to a profound inhibition of T cell activation. Since glutathione levels in rheumatoid arthritis are known to be abnormally low, these results may explain how auranofin can act to suppress the immunological processes leading to rheumatoid arthritis. In addition, this study further demonstrated the close link which exists between auranofin action and glutathione metabolism in lymphocytes. PMID- 8242759 TI - Intracellular induction pathways for CD23 antigen and surface immunoglobulins in human tonsillar B cells: the roles of protein kinase C and tyrosine kinase mediated signals. AB - The expression of CD23 on human tonsillar B cells is increased following treatment with interleukin 4 (IL-4) or 12-O-tetradecanoylphorbol 13-acetate (TPA), while that of surface immunoglobulins (sIgs) is increased by IL-4 but decreased by TPA. This suggests that the signaling by these effectors may result from distinct second messenger-generating systems. In this study, we attempted to elucidate the signal transduction pathways responsible for the expression of CD23 and sIgs by using different protein kinase C (PKC) and tyrosine kinase (TK) inhibitors. Our results showed that B cells expressed varying amounts of sIgs depending on different activators and inhibitors. Sphingosine, a PKC inhibitor, almost completely reversed the TPA-induced decrease in sIgM and sIgD expression. Other PKC inhibitors, e.g., H7 and staurosporine, had similar but less profound effects. In comparison, the up-regulation of CD23 by IL-4 and TPA was only partially blocked by these PKC inhibitors. TK inhibitors, such as herbimycin A and genistein, decreased both the IL-4- and TPA-induced CD23 expression by 50 80%, but had modest effects on sIgs expression. These findings indicate that CD23 and sIgs expression is regulated by independent pathways; PKC is important for the regulation of sIgs expression while the signals through TK pathways might play the major role in CD23 expression. PMID- 8242760 TI - The role of antigen-presenting cells in the regulation of delayed-type hypersensitivity. II. Epidermal Langerhans' cells and peritoneal exudate macrophages. AB - A new mechanism for regulation of delayed-type hypersensitivity (DTH) was investigated. The subcutaneous injection without adjuvant of syngeneic epidermal Langerhans' cells (LC) pulsed with keyhole limpet hemocyanin (KLH) into BALB/c mice gave rise to DTH upon challenge of the ear with the same antigen. When such cells were transferred intravenously, DTH did not occur, although the titer of anti-KLH antibodies was high. Peritoneal exudate macrophages (M phi) pulsed with KLH gave rise to neither DTH nor antibody production. The intravenous transfer of KLH-pulsed LC into mice immunized subcutaneously with KLH in complete Freund's adjuvant at the same time (in the sensitization phase) had a suppressive effect on DTH in an H-2-restricted way. M phi did not have immunoregulatory effects. When radiolabeled LC and M phi were transferred intravenously, they migrated into the spleen, but when they were transferred subcutaneously, they stayed in the skin or migrated into the lymph nodes. In splenectomized mice immunized with KLH, the intravenous transfer of LC pulsed with KLH neither caused the production of anti-KLH antibodies nor suppressed DTH. When Ia was expressed on the surface of M phi, the cells could present antigens, as LC could. These findings suggest that the anatomic sites at which an antigen is presented (i.e., spleen or draining lymph nodes) rather than the kind of cell that first presents an antigen to the immune system is important in deciding whether the immune response that takes place is DTH or antibody production. PMID- 8242761 TI - Effect of CD4 engagement on CD4-T cell receptor complexes. AB - To detect the presence of CD4-T cell receptor (TCR) complexes, we previously developed a flow cytometric method for measuring singlet-singlet energy transfer on human T cells labeled with fluorescein isothiocyanate-conjugated anti-CD4 and tetramethylrhodamine isothiocyanate-conjugated anti-TCR. Using the same procedure, we have now studied changes in the expression of CD4, TCR, and CD4-TCR complexes following CD4 engagement. Ligation of the D3 domain with OKT4, or the D1 domain with anti-Leu3a, induced CD4 and TCR down-regulation, while ligation of the D1 domain with gp120 did not. OKT4 caused a transient decrease in CD4-TCR association over 1 hr at 37 degrees C, while anti-Leu3a caused a steady-state decrease. In contrast, gp120 decreased CD4-TCR association mainly at 0 degrees C, rather than at 37 degrees C. Such alteration in CD4-TCR assembly may underly anti Leu3a- and gp120-mediated inhibition of T cell antigen recognition and account for the negative effect of CD4 ligation on TCR-triggered responses. PMID- 8242762 TI - Phenotypic characterization of antibody-secreting cells in the liver. AB - We compared the phenotypic characterization of antibody-secreting cells in the liver, the spleen, and Peyer's patch that had been subjected to various treatments in mice. The highest proportion of IgA-secreting cells was found in the liver mononuclear cells (MNC). They contained IgM-secreting cells in similar proportions to those in Peyer's patch but at lower proportions than in the spleen. In immunized mice, levels of IgG-secreting cells in the spleen and the liver were about 10 times higher than those in normal mice. The distribution pattern of immunoglobulin-secreting cells in Peyer's patch was virtually unchanged by any treatment. In the normal mice there were no significant differences among these organs in the distribution of the surface immunoglobulin phenotype in MNC. IgA-secreting cells in Peyer's patch secreted more antibodies per cell than those in the spleen or the liver. PMID- 8242763 TI - Expression of the gene for tumor necrosis factor-beta but not for tumor necrosis factor-alpha is impaired in tumor-bearing mice. AB - Spleen cells from mice bearing progressively growing syngeneic sarcomas are immunologically hyporeactive and respond by significantly decreased proliferative response to stimulation with mitogens and cytokines. Here we show that these hyporeactive cells synthesize, after mitogen stimulation, comparable amount of mRNA for tumor necrosis factor (TNF)-alpha as do cells from control mice. However, stimulated spleen cells from the same tumor-bearing mice produce considerably less mRNA for TNF-beta than cells from control mice. These observations were further confirmed using purified peritoneal macrophages and enriched splenic T cells. The results thus demonstrate a distinct regulation of expression of genes for TNF-alpha and TNF-beta, two functionally very similar cytokines, and simultaneously a selective impairment of T-cell function in the course of growth of syngeneic tumors in mice. PMID- 8242764 TI - Retinoic acid promotes proliferation and induces expression of retinoic acid receptor-alpha gene in murine T lymphocytes. AB - We have studied the direct effect of retinoic acid on the antigen-specific proliferative response of a T lymphocyte clone and have investigated the related expression of the retinoic acid receptor-alpha gene (RAR-alpha). Retinoic acid provided direct growth support, in a dose-dependent manner, for hen egg white lysozyme (HEL)-activated T lymphocytes in serum-free medium. The increase of T lymphocyte proliferation supported by retinoic acid was dose dependent, and best effects were achieved at physiological concentrations (10(-9)-10(-7) mol/liter). Expression of RAR-alpha mRNA in antigen-stimulated T lymphocyte cultures was upregulated by retinoic acid in a dose-dependent manner; maximal expression occurred within 6 hr of culture and in response to 10(-9)-10(-7) mol/liter retinoic acid. Lower expression of RAR-alpha was attained by antigen or mitogen (concanavalin A) in the absence of externally added retinoic acid. The inclusion of retinoic acid at the time of HEL processing led to increased T lymphocyte proliferation, provided antigen presenting cell cultures had been maintained for 16 hr. Our results indicate that retinoic acid and RAR-alpha might function in T lymphocytes in a manner similar to that described in other cell systems, namely as ligand-inducible transcriptional enhancer factors. PMID- 8242765 TI - The 4F9 antigen is a member of the tetra spans transmembrane protein family and functions as an accessory molecule in T cell activation and adhesion. AB - In this report, we describe a 43- to 50-kDa protein, which may function as a costimulatory molecule for full activation of human T cells. This Ag, defined by a mouse monoclonal antibody (mAb) anti-4F9, is primarily distributed on "helper/inducer" or "memory" CD4+CD45RO+ subset. Like mAbs against many other accessory/costimulatory molecules, coimmobilization of anti-4F9 with anti-CD3 resulted in synergistic T cell proliferation. In addition, immobilized anti-4F9 on plastic plates induced T cell spreading characterized by the development of prominent dendritic processes. A cDNA encoding the 4F9 Ag was isolated from a cDNA library constructed from PHA/PMA-activated T cells using a COS cell expression system. The sequence of the cDNA and a homology search revealed that the 4F9 Ag was identical to R2, a molecule recently cloned by subtractive hybridization. The 4F9/R2 Ag belongs to a newly identified supergene family (tetra spans transmembrane protein family) characterized by four putative transmembrane domains which are highly conserved between the members of this family. Based upon the phenotypical and functional studies described here, we propose that the 4F9 Ag is an integral membrane protein which can transmit signals involved in T cell proliferation and adhesion. The preferential distribution of this molecule on the CD4+CD45RO+ subset of T cells may contribute to the distinct activation profile and functional repertoire of these cells. PMID- 8242766 TI - Characterization of MHC induction by Mycoplasma fermentans (incognitus strain). AB - Mycoplasma fermentans (incognitus strain) is a recently identified new human pathogen and suspected cofactor in acquired immune deficiency syndrome. Because this organism appears to exert strong immunosuppressive properties of its own, we decided to investigate whether it was capable of inducing MHC class II expression, as we have observed for other species of mycoplasma. In this report we demonstrate that M. fermentans (incognitus strain) is capable of producing factors that increase MHC class II expression as well as MHC class I expression on the myelomonocytic cell line, WEHI-3 cells. We also present data showing that these mycoplasmal factors induce small, although significant, increases in MHC class I and II antigens on a mouse glioma cell line, G26-20, and MHC class II expression on the human monocyte cell lines, U-937 and HL-60. Using nuclear run on analysis, we show that the mycoplasma-induced increase in MHC expression is at least partially due to an increase in transcription of the MHC genes. Furthermore, we show that the factor that mediates this activity is sensitive to protease treatment, indicating that it is, at least in part, protein. These results demonstrate that M. fermentans (incognitus strain) is capable of modulating the expression of immunologically important MHC genes in both murine and human cell lines, which may prove to be an important factor in the pathogenesis of this organism. PMID- 8242767 TI - Induction of mouse thymocyte apoptosis by inhibitors of tyrosine kinases is associated with dephosphorylation of nuclear proteins. AB - Incubation of mouse thymocytes with the protein tyrosine kinase inhibitors herbimycin A and methyl-2,5-dihydroxycinnamate induced a decreased and altered profile of nuclear phosphotyrosine proteins in parallel with an increase in internucleosomal DNA fragmentation and cell death dose-dependently. No change in the profile of cytoplasmic phosphotyrosine proteins was observed. DNA fragmentation was dependent on the synthesis of RNA and protein, suggesting that the inhibition of tyrosine phosphorylation of the nuclear proteins induces apoptosis. DNA fragmentation was enhanced by simultaneous incubation with phorbol esters capable of activating protein kinase C. Genistein, another inhibitor of protein tyrosine kinase, induced DNA fragmentation more rapidly than herbimycin A, but there was no predominant alteration of phosphotyrosine proteins in early incubation, suggesting that genistein may induce apoptosis by a mechanism other than direct inhibition of protein tyrosinekinase activity. PMID- 8242768 TI - Direct demonstration of the CD27 molecule involved in the negative regulatory effect on T cell activation. AB - CD27 is a member of the nerve growth factor/tumor necrosis factor receptor family and appears to play an important role in the process of T cell activation. We have previously shown that perturbation of the CD27 molecule by anti-1A4 (CD27) mAb inhibits T cell proliferation induced by various stimuli. To further determine the molecular basis of this inhibition of T cell proliferation by anti 1A4 mAb, we have established CD27-transfectant lines. Introduction of the human CD27 cDNA to ovalbumin-specific I-Ad-restricted murine T cell hybridomas significantly reduced their antigen-specific IL-2 production. In addition, treatment of these human CD27 transfectants with an anti-CD27 mAb (anti-1A4) further reduced their IL-2 production. These results provide direct evidence that CD27 plays a crucial role in the process of T cell activation. PMID- 8242769 TI - Induction of unresponsiveness in adult rats by immunodominant and nondominant peptides. AB - Intravenous (i.v.) administration of antigens in aqueous solution is an efficient procedure for inducing unresponsiveness in adult animals. Using this procedure, we have compared the unresponsiveness-inducing capacity of an immunodominant and a nondominant peptide derived from the sequence of a retinal antigen, interphotoreceptor retinoid-binding protein (IRBP). The immunodominant peptide, R16, is highly immunogenic in the Lewis rat, while the nondominant peptide, R4, is much less immunogenic and requires higher doses to induce immune response. Pretreatment of Lewis rats with peptides R4 or R16 effectively induced unresponsiveness, shown by the reduced lymphocyte responses following the challenge with the same peptide in adjuvant emulsion. A correlation was found between the doses of each peptide needed to induce immunity or unresponsiveness: both of these doses had to be higher for peptide R4 than for R16. Remarkably, the minimal unresponsiveness-inducing doses of both peptides were lower than those used for the challenge. Pretreatment with the immunodominant peptide, R16, readily inhibited the response to this peptide by lymphocytes from rats challenged with whole IRBP and excessive doses of R16 even affected the response of these rats against whole IRBP. In contrast, R4, which is not recognized by cells of rats challenged with whole IRBP, had no effect on the responses of these rats. The data thus underscore the relationship between immunodominance and unresponsiveness-inducing capacity of peptides and support the notion that the processes of unresponsiveness induction are related to those that produce immunity. PMID- 8242770 TI - The effects of temperature and oleic acid on murine memory and virgin T cell activation: interleukin-2 secretion and interleukin-2 receptor expression. AB - Previous studies established that low in vitro temperatures (27 degrees C, termed nonpermissive) suppressed murine primary thymus-dependent (TD), but not primary thymus-independent (TI) or secondary TD, antibody responses. Suppression was rescued by the addition of oleic acid (18:1), recombinant interleukin (rIL)-2 and/or rIL-4, but not rIL-1. These observations suggested that low temperatures suppress the functions of virgin T cells (Tv) but not those of memory T cells (Tm), B cells, or accessory cells and that hypothetically 18:1 may rescue suppressed responses by altering the membranes of Tv cells, allowing them to proliferate and subsequently secrete interleukins. In this study negatively selected Tm and Tv cells were stimulated with various mitogens at 37 or 27 degrees C. The results indicated that proliferation was suppressed at 27 degrees C to all mitogens tested. The subsequent addition of 18:1 induced proliferative responses at 27 degrees C to both concanavalin A (Con A) and a combination of 12 O-tetradecanoylphorbol 13-acetate (TPA) and calcium ionophore (A23187), but not to phytohemagglutinin-P (PHA). Both Tm and Tv cells showed significant secretion of IL-2 and expression of IL-2 receptor (IL-2R) at 27 degrees C in response to all mitogens, irrespective of proliferation, and the subsequent addition of 18:1 caused little or no change in the levels of IL-2 secretion or IL-2R expression. These findings indicate that suppression of Tm and Tv cell proliferative responses occurs irrespective of IL-2R expression and, unlike antibody production, of IL-2 secretion. Furthermore, it appears that 18:1 can synergistically act with Con A or TPA/A23187, but not PHA, in activating Tm and Tv cells to induce proliferation at 27 degrees C. These findings suggest differences in signal transduction mechanisms between these various mitogens. PMID- 8242771 TI - Lymphokine expression in granulomas of Schistosoma mansoni-infected mice. AB - Lymphokine mRNA for IL-2, IL-4, IL-5, IL-10, and IFN-gamma was identified in total RNA, extracted from granuloma and spleen cells of Schistosoma mansoni infected mice. The specific mRNA in these preparations was identified after reverse transcription and polymerase chain reaction amplification. In some experiments, spleen cells were incubated in the presence of soluble egg antigens or mitogen before RNA extraction. Also, the secretion of these lymphokines from cells maintained in vitro was measured by ELISA or bioassay. The spleen cells from either uninfected or infected mice constitutively released no detectable cytokines and expressed only IL-10 mRNA prominently. Mitogen induced spleen cells from either source to express IL-2 and IFN-gamma mRNAs and their corresponding proteins. Also, spleen cells from noninfected mice stimulated with mitogen expressed IL-4 mRNA, but not IL-5 mRNA, and no detectable protein products. Yet, IL-5 was inducible in spleen cells from infected animals. Soluble egg antigens induced spleen cells from infected mice to prominently express a variety of cytokine genes and produced large amounts of cytokine proteins, but spleen cells from uninfected mice did not respond. Unlike the spleen, the granulomas constitutively expressed mRNAs for all the cytokines examined. Yet, there was limited expression of IL-2 mRNA as compared to that of other cytokines. Except for IL-5, unfractionated granuloma cells constitutively released cytokines at low or undetectable levels. Soluble egg antigens or mitogen enhanced the production of all cytokines to different extents. These observations indicate that schistosome granulomas and spleens express both Th1 and Th2 cytokines. The various lymphokines display complex differences in both lymphokine mRNA and product expression. These results suggest that the production of granuloma lymphokines is governed selectively, possibly by a variety of undetermined regulatory pathways. PMID- 8242772 TI - Interferon-gamma-stimulated uptake and turnover of linoleate and arachidonate in macrophages: a possible pathway for hypersensitivity to endotoxin. AB - When P388D macrophage cells were exposed to [14C]linoleic acid, phosphatidylcholine (PC) phosphatidylethanolamine (PE), phosphatidylinositol (PI), and triacylglycerol (TAG) were major labelled lipid classes. PC was the major labelled phospholipid class at shorter incubation times with PE and PI becoming proportionally better labelled with time. Interferon-gamma (IFN-gamma) stimulated the labelling of all phospholipid classes at the expense of triacylglycerol (TAG) labelling whether it was added coincidently to the [14C]linoleic acid or after a prelabelling period. A similar pattern of labelling of all major phospholipid classes by interferon-gamma in J774.2 macrophage cells was also observed. Interferon-gamma also exerted a stimulatory effect on incorporation of [14C]arachidonic acid into the phosphatidylinositol fraction of the membrane phospholipids. However, uptake of [14C]stearic acid was not different in control compared with IFN-gamma-activated cells. Uptake of linoleic acid into the plasmalogen fraction of PE was also considerably enhanced in IFN gamma-treated cells. The results suggest that IFN-gamma has a direct effect on the activity of enzymes controlling fatty acid turnover in phospholipids. This altered uptake and turnover of unsaturated fatty acids may have important consequences for the subsequent activation of macrophages by endotoxin. PMID- 8242773 TI - Regulation of plasminogen activator inhibitor-1 levels in human monocytes. AB - The regulation of PAI-1 synthesis by elutriation-purified human monocytes was studied in vitro and compared to that for PAI-2. PAI-1 formation, as measured by ELISA, was upregulated by TGF-beta (> or = 1 ng/ml) and surprisingly down regulated by LPS (100 ng/ml), particularly in the presence of TGF-beta; LPS elevated PAI-2 levels (ELISA) while TGF-beta reduced its basal levels and those in LPS-treated cultures. Concomitant changes in mRNA expression occurred. The glucocorticoid dexamethasone (10(-7) M) elevated PAI-1 and acted in concert with TGF-beta in this regard at both the antigen and mRNA levels; interleukin-4 (IL-4) (250 pM) failed to mimic the steroid in its regulation of PAI-1 formation. Since monocyte/macrophage PA activity is likely to be important in tissue remodeling and cell migration at sites of inflammation and in fibrinolysis, it is proposed from these studies that PAI-1, as well as the usually considered PAI-2, may be involved in the negative control of PA activity in this cell type. The synthesis of each PAI appears to be independently regulated. PMID- 8242774 TI - Activation of murine lymphocytes by exogenous phosphatidylethanolamine- and phosphatidylcholine-specific phospholipase C. AB - Enzymes of the protein kinase C (PKC) family have a decisive role in the activation process of T cells. Under physiological conditions, PKC undergoes activation by diacylglycerol (DAG) which is transiently produced following activation of an antigen receptor-coupled phosphatidylinositol 4,5-bisphosphate (PIP2)-specific phospholipase C (PLC) and hydrolysis of PIP2. In vitro activation of distinct PKC isoenzymes is differentially regulated by various synthetic DAGs and membrane phospholipids. We tested whether exogenous PLC obtained from Bacillus cereus can affect PKC-dependent functions in mouse lymphocytes. The results indicated that B. cereus PLC, which preferentially hydrolyzes phosphatidylethanolamine (PE) and phosphatidylcholine (PC), induces the expression of functional cell surface IL2 receptors and that, in the presence of exogenous IL2, it leads to cell proliferation. The PE/PC-PLC reversed the inhibitory effect of PC on enzymatic activity of T cell-derived PKC in vitro. Furthermore, it augmented the activity of PKC over the background level suggesting that the PC-derived DAG may function as a positive regulator of T cell specific PKC isoenzymes. PMID- 8242775 TI - The RT6 rat lymphocyte alloantigen circulates in soluble form. AB - RT6 is a rat maturational lymphocyte alloantigen that appears to subserve important immunoregulatory functions. The lymphopenic diabetes-prone BioBreeding (BB)/Worcester rat is severely deficient in RT6+ T cells and develops spontaneous autoimmune diabetes mellitus. Transfusion of RT6+ T cells prevents the disease. Conversely, in vivo immune elimination of RT6+ T cells from the diabetes resistant line of BB rats induces diabetes and thyroiditis. RT6 protein is expressed in two allotypic forms, each linked to the cell surface by a phosphatidylinositol (PI) anchor. The mechanism by which RT6+ T cells exert their regulatory function is not known, nor is the function of the RT6 protein defined. In this study, we investigated the possibility that, like other PI-linked proteins, RT6 also exists in a soluble form in the circulation. Using standard biochemical procedures we observed: (i) Soluble RT6 circulates in readily detectable amounts in all rat strains studied. (ii) The diabetes-prone BB rat circulates less RT6.1 than does any other strain, including the coisogenic diabetes-resistant line. (iii) Injections of monoclonal anti-RT6.1 antibody rapidly eliminate soluble RT6 from the circulation of diabetes resistant BB rats. The existence of a soluble form of a protein associated with immunoregulatory T cells suggests the possibility that soluble RT6 itself might possess immunomodulatory properties. PMID- 8242776 TI - Reduced activation of transcriptional factor AP-1 among peripheral blood T cells from elderly humans after PHA stimulation: restorative effect of phorbol diesters. AB - In the present study, we evaluated if aging influences the activation and characteristics of transcriptional factor AP-1 in human T cells. Using gel mobility shift assays, the activation of AP-1 was quantified in peripheral blood T cells from 11 elderly (mean 74 year) and young (mean 33 year) subjects following stimulation with PHA, PMA, or PHA plus PMA. The results showed that the activation of AP-1 was significantly reduced in PHA-stimulated T cells from the group of elderly subjects when compared to T cells from young subjects (P < 0.05). Even though PHA-stimulated T cells from 8 of the elderly subjects had pronounced impairments in the activation of AP-1, additional signals provided by costimulation with PMA frequently restored AP-1 activation to more normal levels. Other experiments demonstrated that the AP-1 complexes expressed by stimulated T cells of elderly and young subjects exhibited similar properties in gel shift assays with competing unlabeled AP-1 oligonucleotides and with blocking antibodies specific for Fos and Jun. Thus, these data suggest that the activation of AP-1 can be reduced in human T cells during aging and that these reductions may often be related to impairments in signal transduction rather than represent an absolute loss in the ability to express AP-1. PMID- 8242777 TI - Digital imaging in craniomandibular practice. PMID- 8242778 TI - Reliability and validity of the pressure pain thresholds (PPT) in the TMJ capsules by electronic algometer. AB - Pressure pain thresholds (PPT) of 39 normal subjects (20 males and 19 females) and 30 female patients with temporomandibular joint (TMJ) capsulitis (21 lateral and 25 posterior capsulitis) were examined by an electronic algometer. In normal subjects, statistically significant correlation coefficients were obtained from the values of intra-examiners and inter-examiners in both lateral and posterior TMJ capsules (p < 0.01). A comparison with data obtained from contralateral sides failed to demonstrate significant differences. Statistically significant differences were found between the PPT of normal female subjects and female patients with capsulitis in both lateral and posterior TMJ capsules (p < 0.01). PMID- 8242779 TI - A comparison of electromyographic activity between anterior repositioning splint therapy and a centric relation splint. AB - The purpose of this study was to compare the electromyographic activity of masticatory muscles (temporal and masseter) with the use of an anterior repositioning splint and a centric relation superior repositioning splint. Twenty six consecutive patients, who referred with the chief complaint of temporomandibular pain and/or headache were selected from one of the author's practices. All these subjects were diagnosed as having internal derangement of the temporomandibular joint. Ten normal subjects were used as controls. Surface electromyographic recordings were taken of each subject prior to the beginning of clinical therapy for the patients. The results show significantly less masseter and temporal muscle activity with anterior repositioning splint therapy compared to the centric relation superior repositioning splint therapy. PMID- 8242780 TI - The immediate effect of the variation of anteroposterior laterotrusive contacts on the elevator EMG activity. AB - A unilateral maxillary splint divided into three occlusal blocks was built for eight healthy young adults. The three blocks allowed the anteroposterior location of the laterotrusive contacts to vary and to distribute the occlusal force over equivalent periodontal surfaces. The ipsilateral masseteric and temporalis electromyogram (EMG) activities were recorded with surface electrodes during maximal voluntary clenching. The elevator EMG activity with the anterior occlusal block in place was significantly lower than with the intermediate or posterior blocks. The reduction for both elevators with the intermediate block in place did not show a significant variation in comparison to the posterior block of the three blocks inserted. The elevator activity with the three occlusal blocks did not differ from that recorded with the posterior block alone. Neurophysiologic and biomechanical explanations are given related to this laterotrusive elevator muscular behavior. PMID- 8242781 TI - Vibration analysis of the temporomandibular joints with meniscal displacement with and without reduction. AB - The vibrations of 102 joints demonstrating meniscal displacement with either early or late reduction (MDR-early/MDR-late) and 70 joints displaying meniscal displacement without reduction either incomplete or complete (MD-incomplete/MD complete) were analyzed and compared to 83 arthrographically normal but symptomatic joints (NID) using electrovibratography (EVG). The total power density of the vibration [I(T)], peak power density [I(max)] and power density at each 50Hz range between O to 600 Hz [I(f)] showed the highest in the MDR-late group followed by the MDR-early group, suggesting that the level of vibration is related to the degree of disk displacement and reduction. The wave characteristic parameters such as the correlation coefficients between I(T) and each I(f) showed higher correlation at higher frequency ranges as the degree of disk displacement progressed, from MDR-early to MDR-late to MD-incomplete. The diagnostic sensitivity of EVG when using I(T) as a determining parameter was 96.6% for the MDR-early group, 91.8% for the MDR-late group, 77.8% for the MD-incomplete group and 57.4% for the MD-complete group with the specificity for the NID group at 75%. PMID- 8242782 TI - Vibrations in the temporomandibular joints in patients examined and treated in a private clinic. AB - Patients indicated for examination, evaluation and treatment of the temporomandibular joint (TMJ) in the Centro de Diagnostico e Tratamento da ATM (CDTATM) were submitted to computerized BioPAK system tests. Special attention was placed on the SonoPAK test (electrovibratography), seeking to evaluate the articular sound problem in differentiated patients for specific TMJ treatment. The percentage of sounds (148 joints, 74 patients) was high (75.67% in the opening movement and 50.00% in the closing movement) which suggests the necessity to give more attention to the TMJ sounds during a clinical exam. And also, if possible, give more attention to the utilization of specific tests to obtain a clear definition of the type of sound the patient presents. PMID- 8242783 TI - Orthopedic jaw movement observations. Part II: The rotational capacity of the mandible. AB - Using computerized axiography, particularly the electronic mandibular position indicator (EMPI), the mandibular opening movement was measured in 86 asymptomatic volunteers and temporomandibular joint (TMJ) patients. Terminal hinge-axis movement and hinge-axis rotation at maximum-guided mouth opening were recorded. The angle of hinge-axis rotation was used as a parameter for both movements, in accordance with the neutral-zero method. This is a well-established technique in orthopedics and is the standard tool for quantitative functional analysis of joints. An average group was defined, 50% being volunteers. The interquartile range for terminal hinge-axis movement in this study was 5.42 degrees-7.41 degrees in the volunteers and 4.73 degrees-7.25 degrees in the patients. The interquartile range for hinge-axis rotation at maximum opening was from 29.09 degrees-34.87 degrees in the volunteers and from 26.7 degrees-35 degrees in the patients. Computerized axiography is a refined tool for analyzing rotational and translational capacities of the mandible. It is a valid and practical method for orthopedic-diagnostic evaluation of mandibular movements and gives objective criteria for diagnosis in accident victims. PMID- 8242784 TI - Measuring the impact of a dental practice on TM disorder symptoms. AB - This article describes a practical, quantitative method of measuring changes in temporomandibular disorder (TMD) symptoms in a dental practice. It applies the TMJ Scale to produce a number of clinically important measures of treatment effectiveness. Those measures include pre- and post-treatment symptom severity, percent of patients improved, level of improvement, and percent of patients converting to non-symptomatic after treatment. All of these outcome parameters were applied to a sample of the practice and to subgroups based on age, sex and problem length. The treated patients were compared to a group of diagnosed but untreated patients. The specific target symptoms of TMD are analyzed including pain, palpation pain, joint dysfunction and limited range of motion of the mandible. In addition, the overall symptom severity of the TM disorder, psychological factors and stress are studied, leading to an assessment of symptom change. The results support the concept that patients with acute problems are more treatment responsive than are patients with chronic problems. This suggests that careful screening and earlier detection of TMD may have an important impact on ultimate treatment outcome. PMID- 8242785 TI - Prevalence of temporomandibular disorder symptoms in a major metropolitan area. AB - An accurate description of the population prevalence of temporomandibular disorders (TMDs) would be of value to scientists, health-policy makers, and clinicians but, to date, various unreplicated epidemiologic methodologies have produced disparate prevalence figures. We report on a telephone survey that sought to describe the prevalence of six TMD-related symptoms among a random, non clinic sample from the Kansas City metropolitan area. The prevalence figures for nocturnal bruxism, diurnal clenching, jaw soreness and joint sounds ranged from 10-19%. Some figures were higher than those from St. Louis, but generally the figures accord well with the results of other prevalence studies. Of the 534 people interviewed, 246 reported one or more of the six TMD-related symptoms. Symptoms were not more prevalent among women than men, but were higher among persons of age 45 and under. Pain was reported more commonly by persons with multiple symptoms. The level of concordance between data from Kansas City and St. Louis supports the validity of the telephone survey method for studying TMD prevalence in non-clinic samples. PMID- 8242786 TI - Articular disk displacement of TMJ due to trauma. AB - One hundred and thirty native American Indian patients undergoing orthodontic treatment for malocclusions were screened for temporomandibular joint (TMJ) abnormalities. These patients were followed over the course of orthodontic treatment and six of these patients suffered a traumatic injury to the TMJ. These six patients who had been previously screened and found to be asymptomatic, developed symptoms. Three were diagnosed with articular disk displacement of the TMJ confirmed by magnetic resonance imaging (MRI). PMID- 8242787 TI - Craniomandibular dysfunction in children and adolescents: a review. AB - The purpose of this paper is to show that temporomandibular disorders manifest as much in children and adolescents as in adults. Objective signs, such as pain in the temporomandibular joint area to palpation or function, were found to be 40.6% on average from ages 6-18 years. Subjective signs reported by patients or their parents, such as clicking, pain to function and muscle tenderness, had an average prevalence of 38.7%. All but one researcher reported no difference between signs and symptoms as related to sex. These same articles concurred that the symptoms in children and adolescents, though less severe, were slightly higher in incidence than the adult population. Since the prevalence of signs and symptoms of temporomandibular disorders is the same in children and adolescents as in adults, more attention needs to be placed on its recognition in this age group. In addition, more research into treatment modalities and possible preventive measures in this age group is required to prevent sequelae in later years. PMID- 8242788 TI - Osteocavitation lesions (Ratner bone cavities): frequently misdiagnosed as trigeminal neuralgia--a case report. AB - The disorder termed osteocavitation lesion has been described in the literature since at least 1976. This disorder has often been misdiagnosed as trigeminal neuralgia or atypical facial pain, and, unfortunately, patients have either continued to suffer or inappropriate treatment or treatments have been prescribed in an attempt to rid the patient of this terrible pain disorder. These symptoms, which can be misinterpreted as trigeminal neuralgia, include a history of undiagnosed facial pain, a history of tooth extraction, the presence of trigger areas and normal radiographic findings. A confirmed diagnosis of osteocavitation lesion can be treated only with surgery. PMID- 8242789 TI - Legal perspectives on TMJ/whiplash. PMID- 8242790 TI - Clinical joint sound analysis. PMID- 8242791 TI - Orofacial physiotherapy after radiotherapy in the head and neck region. PMID- 8242792 TI - Isolation of a new species of Physarum lysophosphatidic acid, PHYLPA, and its effect on DNA polymerase activity. AB - A new species of lysophosphatidic acid was isolated from myxoamoebae of a true slime mold, Physarum polycephalum, and structural studies were performed. The purified substance was subjected to nuclear magnetic resonance spectroscopy (NMR), infrared spectroscopy (IR), fast atom bombardment mass spectroscopy (FAB/MS), alkaline hydrolysis and tandem mass spectroscopy (MS/MS), and the results suggested this substance to be lysophosphatidic acid composed of a cyclic phosphate and cis-11,12-methylene octadecanoic acid. The effects of the LPA on DNA polymerases were studied and compared with the effects of PHYLPA, which had been isolated as a specific inhibitor of eukaryotic DNA polymerase alpha (6). It showed a specific inhibitory activity on eukaryotic DNA polymerase alpha, but no activity on the repair-type, or mitochondrial DNA polymerases. PMID- 8242793 TI - Folimycin (concanamycin A), a specific inhibitor of V-ATPase, blocks intracellular translocation of the glycoprotein of vesicular stomatitis virus before arrival to the Golgi apparatus. AB - Folimycin (concanamycin A) specifically inhibited vacuolar-type ATPase as far as examined. Folimycin blocked excretion of the glycoprotein (G protein) of vesicular stomatitis virus into the medium and, instead, G protein was accumulated intracellularly. The intracellularly accumulated G protein electrophoresed a little faster than mature one. The N-glycan of the G protein was endoglycosidase H-sensitive, and terminal galactose and N-acetylglucosamine were not detected essentially on sequential digestion with exoglycosidases, indicating that processings known to occur in the Golgi apparatus do not take place in the presence of folimycin. The oligosaccharide chain of the G protein was determined to have a composition of Man8GlcNAc2 as analyzed by Bio-Gel P-4 column chromatography and high-performance liquid chromatography following digestion with alpha- and then with beta-mannosidase. Activities of mannosidase I and glycosyltransferases prepared from baby hamster kidney cells were not inhibited as far as examined, indicating that the incompleteness of the N glycosidic chain in folimycin-treated cells is not caused by inhibition of processing enzymes. Taken together these observations suggest that folimycin blocks the intracellular translocation of G protein before the step of trimming by mannosidase I which is confined to the cis compartment of the Golgi. The intracellular localization of G protein as revealed by fluorescence microscopy was in good accordance with this assumption. PMID- 8242794 TI - Cytoskeletal active drugs modulate signal transduction in the protein kinase C pathway. AB - The cytoskeletal network of cells is postulated to play a role in the signal transduction pathways of growth promoting stimuli. We show here that cytoskeletal active drugs modulate the mitogenic signal transduction pathway of the tumor promoter TPA in 3T3-L1 cells. Compounds which act on microtubules (vinblastine sulfate) and microfilaments (cytochalasin B) have opposite effects on DNA synthesis. Vinblastine sulfate leads to stimulation, whereas cytochalasin B causes potent inhibition of DNA synthesis in response to TPA. These drugs are cell cycle specific and apparently exert their regulatory action distal to activation of protein kinase C by TPA. The expression of four genes necessary for DNA synthesis in response to tumor promoters was examined: two nuclear proto oncogenes (c-myc and c-fos), a transcription factor (c-jun/AP-1) and a key enzyme involved in polyamine synthesis (ornithine decarboxylase). c-jun mRNA levels are not modulated during the action of cytoskeletal disrupting drugs on TPA-mediated mitogenesis, whereas c-myc and c-fos mRNA levels are similarly enhanced. Expression of ornithine decarboxylase mRNA and protein is increased by vinblastine sulfate but decreased by cytochalasin B in TPA treated cells. These data indicate that changes in cytoskeletal organization may play a role in regulating the levels of an enzyme critical for DNA synthesis. PMID- 8242795 TI - Purification and characterization of a lectin-like substance from silkworm faeces. AB - To obtain biologically active substances that are useful in the study of cell structures and functions, we examined several biological activities of an extract of silkworm faeces with hot buffer. We detected a lectin-like substance in the extract and purified it. When the extract of silkworm faeces was added to a culture of quail myoblasts transformed with temperature-sensitive Rous sarcoma virus (QM-RSV cells), the plated cells which were cultured at 35.5 degrees C (the permissive temperature for RSV that suppresses myogenic differentiation) became detached from the dishes and the cells in suspension aggregated. However, when the same amount of extract was added to plated QM-RSV cells cultured at 41 degrees C, which is a nonpermissive temperature for RSV, the cells did not become detached. Other kinds of plated cells examined also did not become detached. Unlike plated cells, nonplated cells of all kinds aggregate in suspension upon the addition of the extract. This active substance was purified by monitoring its induction of cell aggregation. The purified substance was found to be a lectin like glycoprotein with an apparent molecular mass of about 60 kDa. Further studies showed that the binding sites of this glycoprotein are sugar chains on the cell surface and that mannose is an epitope. PMID- 8242796 TI - Difference in laminin expression between high and low metastatic cell clones derived from murine Lewis lung carcinoma. AB - We investigated the expression of laminin in two cell lines with different metastatic potentials established from murine Lewis lung carcinoma. Immunostaining of the cells with anti-laminin antibody and Northern blot analysis of laminin mRNA demonstrated that the high metastatic clone expressed less laminin than the low metastatic one. In contrast, expressions of 67 kDa-laminin receptor were at similar levels between these two lines. These findings show the possibility that endogenous laminin may contribute to the difference in metastatic properties in the murine Lewis lung carcinoma cell lines examined. PMID- 8242797 TI - Activation of the complement system by the capsule of Cryptococcus neoformans. AB - In vitro studies indicate that encapsulated cryptococci are among the most powerful particulate activators of the complement system reported to date. The capsule itself is the site at which activation occurs. Activation occurs solely via the alternative complement pathway. Initiation occurs at apparently random focal sites in the capsule that expand through alternative pathway amplification to fill the capsule with C3 fragments. The C3 fragments are rapidly converted to iC3b, suggesting that phagocyte receptors for iC3b will be important in phagocytosis of the yeast. There is abundant evidence that a similar form of activation occurs in vivo during a cryptococcal infection. Cryptococcemia in humans and experimental animals is accompanied by a depletion of serum complement levels. Studies with complement deficient guinea pigs and mice indicate that the complement system plays an essential role in resistance to cryptococcosis. It is likely that the complement system contributes to host resistance by opsonization of the yeast to facilitate attachment and ingestion by phagocytic cells as well as by releasing chemotactic fragments of the complement cascade which contribute to the inflammatory response. The absence or unavailability of a functional complement system in the central nervous system may account in part for the predilection of the yeast for the brain. PMID- 8242798 TI - Host-parasite relationship in paracoccidioidomycosis. PMID- 8242799 TI - Treatment of coccidioidomycosis. AB - This review has traced chemotherapy of coccidioidomycosis from its unsuccessful beginnings through the present time. Although in vitro susceptibilities give initial impressions of activity of many drugs, and although animal studies identify the diseases targeted for initial trials, ultimately it is the patient in whom the benefits and risks of a new agent must be weighed. For this reason, considerations have been limited to clinical studies, with no review of animal data or immunologic forms of therapy such as transfer factor. In this review, amphotericin B has been found to be the first active antifungal agent in coccidioidomycosis. Although responses clearly occurred, even in meningitis, the vagaries of disease and investigators prevented a really hard assessment of how many patients really were "cured" versus how many relapsed again and again. Responses were given in broad ranges and the term "remission" came to be considered a more accurate definition for this illness than "cure." The azole antifungals, though fungistatic, gave us, for the first time, the ability to treat a patient indefinitely with an oral preparation; the advantages of this cannot be overstated. Although all of these drugs act by similar mechanisms, the differences in potency, pharmacokinetics and toxicities have brought fluconazole and itraconazole to the fore. Which of the two is superior (if either is) cannot be defined with the limited data available. The role of SCH39304 is, at present, unclear. Other agents such as nikkomycin and modified polyenes may enter a role in clinical evaluation of coccidioidomycosis, but for the next five years the drugs described in the preceding pages, or combinations, are likely to constitute the pharmaceutical arsenal against C. immitis. While we now have much more to offer the heirs of Domingo Escurra and Jose Furtado-Silveira than gentian violet and carbolic acid, we still have response rates of less than 75% for extrameningeal disease and we have not yet identified a drug that will guarantee a cure without later relapse. There is still a way to go. PMID- 8242800 TI - A clinician's view of blastomycosis. AB - In summary, blastomycosis is a rarely reported but important infection in certain endemic areas. Better understanding of the number with subclinical infection is dependent on studies with more reliable and specific immunologic testing. The illness may mimic several other conditions but particularly neoplastic disease. Therefore, fungal cultures and smears, which will allow a firm diagnosis, should be obtained during invasive procedures aimed at the diagnosis of cancer. Therapy of blastomycosis has been broadened with the addition of ketoconazole. Itraconazole appears to have even greater efficacy while fluconazole remains to be confirmed as an effective agent for this infection. The role of amphotericin B or itraconazole depends on comparative clinical trials. PMID- 8242801 TI - The presence of dermatophytes in the environment and on healthy looking skin: their significance as a cause of disease in Japan. PMID- 8242802 TI - Entomophthoromycosis. PMID- 8242803 TI - Serodiagnosis of deep-seated fungal infections. PMID- 8242804 TI - Antigenic variability of Candida albicans cell surface. PMID- 8242805 TI - Pathogenesis of dermatophytosis. PMID- 8242806 TI - Infections due to Trichosporon species: new concepts in mycology, pathogenesis, diagnosis and treatment. PMID- 8242807 TI - [Clinical application of the forced inspiratory volume-time curve. A preliminary study]. AB - The forced inspiratory volume-time curve in 25 health subjects with normal ventilation function and 25 patients with COPD were determined, and MMIF, FIV were calculated from the curves. The PImax was measured in the same 50 subjects by modified Blac method sequentially. A close relationship between MMIF, FIV and PImax was found. This result indicated that MMIF and FIV could be indeed for evaluation of strength of respiratory, muscles. Additional observations of MMIF, FIV and PImax in 50 patients with COPD before and after inotropic drugs, aminophylline and CNB, further proved the validity of MMIF and FIV. PMID- 8242808 TI - [Clinical study on patients with lateral position test (LPT) for determining the relative function on each lung]. AB - Comparing the lateral position test (LPT) on 20 patients of lung cancer with bronchospirometry of tracheal intubation, the authors found that the VC, oxygen uptake were in high positive correlation, while VE were in moderate positive correlation. Therefore, they conclude that LPT could substitute the complicated tracheal intubation for determining the relative function of each lung before the total pneumonectomy. PMID- 8242809 TI - [Immunohistochemical investigation on the expression of glutathione S transferases (GSTS) in lung cancer]. AB - Paraffin-embedded tissue sections from 105 cases of human lung cancer were stained for four isozymes of GSTs by immunohistochemical PAP technique. Of the 16 cases of small cell lung cancer (SCLC) examined 14 were negative for all GST individuals, whereas one undifferentiated squamous cell type and one oat cell type treated with chemotherapy before operation were positive for GST-pi. The total positive rates of GST-pi, GSTs and GST-mu in 89 cases of non-SCLC were 75.3%, 13.5% and 9.7% separatively. Among them squamous cell carcinoma were stained positively for GST-pi in 93.5%, GSTs in 9.7% and GST-mu in 6.5%, while adenocarcinoma were in 69.7%, 15.2% and 6.1% respectively. The expression of GST pi was weakened corresponding with the decreased degree of differentiation of cancer cell. GST-alpha was not detectable in all specimens studied. Ultrastructure location of GST-pi sites in 11 cases of non-SCLC was detected mainly on lysozymes and heterochromatin in cancer cell by transmission electronic microscopy utilizing colloidal gold labelled anti-GST-pi antibody. These results suggest that GST-pi may be an useful marker for differential diagnosis in histopathology and intrinsic sensitivity to anticancer drugs of lung cancer. GST mu was expressed in some types of lung cancer with low positive rate and its usage as a marker needs further investigation. PMID- 8242810 TI - [Immunohistochemical study of lung tissues from patients with idiopathic pulmonary fibrosis]. AB - Immunohistochemical analysis using 24 different monoclonal and polyclonal antibodies was applied to lung biopsy and autopsy material from 9 patients with idiopathic pulmonary fibrosis (IPF) and 6 patients without lung disorders as control. Lymphoid, non-lymphoid cells and their subsets as well as extracellular matrix (fibronectin, laminin and collagens type I, III and IV) were observed in lung interstitium. Our results suggest that local B and T lymphoid immune responses are occurring in IPF. Extracellular matrix alterations are closely linked with the local immune responses. PMID- 8242811 TI - [The diagnosis and therapy of 6 cases of pulmonary alveolar proteinosis]. AB - 6 cases of pulmonary alveolar proteinosis were presented. No symptom or physical finding is specific for diagnosis. The final diagnosis can only be documented by patho-morphological examination. PAS positive alcin blue negative lipoprotein in alveolar spaces is the characteristic finding. By using folley's catheter, the authors developed a modified method for pulmonary lavage, and 3 patients were treated effectively. PMID- 8242812 TI - [Thoracoscopy in pleural mesothelioma]. AB - 19 patients suspected pleural mesothelioma were performed thoracoscopy. The overall diagnostic rate was 94.7% (18/19 cases). The macroscopic findings of 18 cases presented solitary, well circumscribed, smooth, rounded masses in 2 localized cases and multiple, clear, yellowish-white and "grape-like" granulations or malignant pleural thickening in 16 diffuse cases. Biopsy was taken in 18 cases under direct vision. The histological diagnosis showed 2 benign fibromatous mesothelioma and 16 malignant mesothelioma. Pleural mesothelioma was easily misdiagnosed as primary lung cancer, tuberculous pleurisy and metastatic pleural tumor. The diagnostic accuracy by cytological study of pleural effusions and needle biopsy of pleura both were low. Thoracoscopy is currently a suitable diagnostic method for pleural mesothelioma. The resection was considered as a proper treatment of choice for localized pleural mesothelioma. We recommend that thoracoscopic talc poudrage followed by chemotherapy on diffuse malignant mesothelioma with pleural effusion could be used to improve the survival rate and to prevent the recurrent effusion. PMID- 8242813 TI - [Polydatin prevents endotoxin-induced acute lung injury in rats]. AB - The effects of polydatin on endotoxin-induced acute lung injury were studied in rats. The lung injury was induced by infusion of inactivated E. coli 30 min after MPS blockade with red cell membrane. It was found that pretreatment with polydatin resulted in marked reduction of lung wet-to-dry weight ratio, lung permeability index (LPI), neutrophil count in BALF and hemoconcentration. The elevation of lung tissue MDA and the decrease in plasma GSH-Px activity were greatly improved with polydatin. There was significant correlation between LPI and lung MDA level (r = 0.741, P < 0.01). Postmortem examination revealed that polydatin attenuated the histopathological changes. We conclude that polydatin protectsral against endotoxin-induced acute lung injury. PMID- 8242814 TI - [The effect of IL-2 on IL-2 production and on the IL-2R expression of spleen cell from mice infected with M. tuberculosis H37Rv]. AB - 36 mice infected with M. tuberculosis H37 Rv were divided into treated group and untreated group at random. 18 infected mice were in treated group with appropriate dose of human recombinant interleukin-2. Normal group consists of 10 normal mice. At 10, 20, 30 day after treatment, IL-2 production and IL-2R expression of mice spleen cells from all three groups were detected. The results showed that: 1) IL-2 and IL-2R level in untreated group was lower than that in treated group and in normal group; 2) In vivo administration of rIL-2 could enhance IL-2 production and IL-2R expression. PMID- 8242815 TI - [Analysis and efficiency-evaluation of case-finding among symptomatic suspects in rural area by smear microscopy]. AB - 216,307 suspects were discovered within the period from Oct. 1st, 1987 to Spt. 30th, 1988 in rural area of 26 counties. The suspect rate was demonstrated to be 2.2%; 182,917 persons accepted smear microscopy among suspects and the smear microscopy rate was 84.6%; 7,046 smear positive cases were detected and the smear positive rate was confirmed to be 3.95% among smear-accepted suspects. Smear microscopy among symptomatic suspects in rural area for infectious case-finding is recommended, because this method costs low, and is more effective and practical. PMID- 8242816 TI - [Clinical and epidemiological significances of L forms of Mycobacterium tuberculosis]. PMID- 8242817 TI - [The changes in erythrocyte calcium is one of the mechanisms of anemia formation in rabbit endotoxin-induced DIC]. AB - The effects of E. coli endotoxin (ET) on human erythrocyte cytosolic free calcium concentration ([Ca2+]i) and membrane calcium pump (Ca(2+)-Mg(2+)-ATPase) activity were observed in vitro. The changes of erythrocyte [Ca2+]i, Ca(2+)-Mg(2+)-ATPase activity, calmodulin activity, the deformability of erythrocytes, membrane protein electrophoresis, and the fluidities of membrane lipids were investigated in the ET-induced disseminated intravascular coagulation (DIC) model to study the significance of erythrocyte factors in the occurrence of microangiopathic hemolytic anemia. The [Ca2+]i of human erythrocytes was 86 +/- 9.2 nmol/L in normal cells, and it was increased to 124 +/- 22 nmol/L, 174 +/- 41 nmol/L and 220 +/- 92.1 nmol/L, respectively, when the erythrocytes were incubated with 0.5 mg/ml, 1 mg/ml and 2 mg/ml ET, respectively. At the same time, the Ca(2+)-Mg(2+) ATPase activities decreased from 1031 +/- 131 nmol/mg.h in normal to 870 +/- 182 nmol/mg.h, 684 +/- 124 nmol/mg.h and 718 +/- 144.4 nmol/mg.h in groups receiving 0.5 mg/ml, 1 mg/ml and 2 mg/ml ET, respectively (P < 0.01). The [Ca2+]i of rabbit erythrocytes was elevated from 76.6 +/- 14.9 nmol/L in normal to 224 +/- 74 nmol/L (P < 0.01), and the Ca(2+)-Mg(2+)-ATPase activity was inhibited from 220 +/- 26.8 nmol/mg.h in normal to 146.1 +/- 30.6 nmol/mg.h during ET-induced DIC in vivo. In addition, erythrocyte deformability decreased and the membrane protein electrophoresis showed changes in the 4.5 KD band area. However, there was no significant difference in the calmodulin activity and fluidities of membrane lipids of erythrocytes in DIC rabbits. These results suggest a possible alternative mechanism for the formation of microangiopathic hemolytic anemia in ET-induced DIC rabbits.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242818 TI - [Histochemical studies of cutaneous lichen amyloidosis using light and electron microscopy]. AB - Ten cases of cutaneous lichen amyloidosis are reported. Amyloid deposits were found to be limited to the papillary dermis. The amyloid masses were often fissured and had an amorphous eosinophilic globular appearance. The protein was confirmed by histochemistry as amyloid-associated protein. The epidermal origin of the amyloid deposit has been confirmed by electron microscopy. The amyloid deposit was of variable island structure, accompanied by filaments. PMID- 8242819 TI - [Distribution of the intermediate filaments in rat Sertoli cells: an electron microscopic study]. AB - The distribution of intermediate filaments (IFs) in the Sertoli cells of rat testis was studied under a transmission electron microscope. The results showed that IFs in Sertoli cells were centrally arranged around the nucleus in a criss cross manner. From the perinuclear region the IFs had widespread extensions and were connected with the following structures: 1) with the mitochondria, Golgi apparatus, endoplasmic reticulum and other subcellular structures; 2) with the desmosome-like and hemi-desmosome-like junctions with adjacent Sertoli and spermatogenic cells; 3) with the microfilaments (MFs) in the ectoplasmic specialization (ES) surrounding the head of elongated stage I-V; spermatids; 4) with the MFs in ES between adjacent Sertoli cells. In conclusion, IFs form a delicate intracellular network in Sertoli cell with its center in the perinuclear region and stretching and connecting with many subcellular structures in the cytoplasm and cell membrane. PMID- 8242820 TI - [Regulation of glycogenolysis against hormones in isolated rat hepatocytes]. AB - cAMP and Ca(2+)-dependent glycogenolytic pathways are regulated by different hormones involved in the activation of glycogen phosphorylase in isolated rat hepatocytes. Dose-dependent glucagon activation of glycogen phosphorylase and lowering of cAMP levels were obtained by vasopressin and angiotensin treatment. Similar time courses of Ca2+ fluxes and phosphorylase activation were induced by vasopressin, angiotensin and phenylephrine. Heterologous down-regulation of hormones was induced in glycogenolysis in hepatocytes. These results could be of significance in physiology studies in vivo. PMID- 8242821 TI - [Studies on the transforming effects of wollastonite on SHE cells]. AB - A transformation system utilizing SHE cells was used to study the carcinogenic effects of Chinese wollastonite, a new mineral fiber. Morphological transformation was observed in SHE target cells treated with either wollastonite or MNNG, a known chemical carcinogen. A single treatment of wollastonite (20 ug/ml) induced typical transformation. The transformation rate of SHE cells induced by MNNG followed by several treatments with wollastonite was higher than that induced by the same dose of MNNG only. Electron microscopic observation showed that the ultrastructure of wollastonite-transformed SHE cells was significantly different than that of normal SHE cells, but similar to that of SHE cells transformed by MNNG. The above results suggest that wollastonite may not be simply an environmental carcinogen, but also a possible promoter. PMID- 8242822 TI - [The preparation and identification of anti-erythropoietin monoclonal antibodies]. AB - By using the partially purified recombinant human erythropoietin (rHuEPO) as an antigen, two hybridoma clones have been obtained that secrete monoclonal antibodies against human erythropoietin. Intrasplenic immunization was successfully used in this experiment. The prepared monoclonal antibodies showed high specificity to human EPO by Ouchterlony, ELISA, dot blot and Western blot tests. They were also found to inhibit the biological activity of rHuEPO in a concentration-dependent manner by an in vitro bioassay. In the future, these monoclonal antibodies will be used as the solid phase antibodies in an enzyme linked immunosorbent assay and as a probe for the purification of native erythropoietin and for the further studies of the hormone and its mechanism of action. PMID- 8242823 TI - [Bronchiolitis obliterans with organizing pneumonia--a case report]. AB - A case of bronchiolitis obliterans with organizing pneumonia was reported. Since the clinical symptoms, chest X-ray and lung function testings are all non specific, open lung biopsy was performed. Final diagnosis was based on characteristic pathologic manifestations. Treatment with corticosteroid was very effective. PMID- 8242824 TI - [Spitz nevus (report of 2 cases)]. AB - This paper reports 2 cases of spitz nevus which appeared on the nose. Both patients were male, aged 3 and 33 years. The histopathologic characteristics showed compound nevus. One patient was treated by CO2 laser and the other by biopsy only. The patients were followed up for 3 and 9 months, respectively, ane no recurrence was found during these period. PMID- 8242825 TI - [Study and application of blood urea nitrogen slide]. AB - Blood Urea Nitrogen (BUN) Reaction Slide was studied and primarily applied in clinical laboratory as a method for BUN test it is ideal of emergency use. BUN Reaction slide is a dry chemical reagent pad, also it is a separation couple-seal system. The slide is divided into two chambers by a membrane, the up one is UN enzymatic reaction area, the bottom one is detector reaction part. This system eliminated interference from blood pH, Hb and bilirubin, only 5 minutes is needed for a single test. The test within day and day-to-day CVs were < 5.5%. Test linearity was up to 1000 mg/L. There was good correlation with BUN enzyme-couple method, r = 0.985, y = 0.91 X + 3.9. The acurrate was better than BUN Strip method. PMID- 8242826 TI - [The metabolism of TMPz in vivo]. AB - The metabolism of tetramethylpyrazine (TMPz) in rats was investigated. Deuterated TMPz and unlabelled TMPz were administered by intragastric perfusion to two rats, respectively. Extracts from urine samples were analyzed within 24h by gas chromatography/mass spectrometry (GC/MS). By applying the ion cluster technique, TMPz and its three metabolites were identified. PMID- 8242827 TI - [Lymphocyte subsets in labial salivary gland of Sjogren's syndrome]. AB - Using the immunohistochemical avidin-biotin-peroxidase complex (ABC) method, lymphocyte subsets in the salivary glands of 15 patients with primary Sjogren's syndrome, 7 patients with secondary Sjogren's syndrome and 4 normal controls were identified by monoclonal antibodies. The infiltrating lymphocytes were mainly T lymphocytes, the majority of which were T helper cells (Leu-3a) in SS groups. The Leu-3a/Leu-2a ratio was increased. These changes were related to the degrees of the disease. The lymphocytes of patients expressed the HLA-DR antigen (Ia). Thus genetic predisposition plays an important role in the disease. A proposed mechanism is that the deficiency of suppressor lymphocytes and the overactivity of helper lymphocytes would result in B cells becoming hyperactive. Multiple autoantibodies produced by the B cells would contribute to tissue damage, and the tissue fragments would in turn further stimulate antibody production. PMID- 8242828 TI - [Effect of lipopolysaccharide on cytosolic free calcium concentration in myeloma cells]. AB - The concentration of cytosolic free calcium was measured using the fluorescent Ca(2+)-indicator quin 2, and the effect of lipopolysaccharide (LPS) on cytosolic free calcium in myeloma cells was studied. Incubation of myeloma cells with LPS elicited a significant increase in cytosolic free calcium, and inhibited the phytohaemagglutinin (PHA)-induced elevation in cytosolic free calcium. Treatment of the cells with phorbol-12-myristate-13-acetate (PMA) failed to affect the concentration of cytosolic free calcium in resting cells, but inhibited the subsequent PHA-induced elevation in cytosolic free calcium. It is suggested that LPS causes alterations in intracellular calcium homeostasis and perturbations in the calcium-dependent signal transduction pathway. PMID- 8242829 TI - [The effects of optical isomers of gossypol on Sertoli cells in vitro]. AB - This study was designed to investigate the effects of gossypol optical isomers on Sertoli cells of 20-day-old Wistar rats in vitro. The dosages used were 10 micrograms/ml of (+/-) and (+) gossypol and 5 micrograms/ml (-) gossypol. After 24 h in culture, we found that different optical isomers induced different degrees of morphological changes in Sertoli cells. The changes in mitochondria were obvious. It was shown that the activity of SDH in mitochondria was reduced, or some enzymatic granules became coarser, after gossypol treatment. Under EM we found that the mitochondria had become swollen, the cristae were arranged irregularly, or even broken or absent. The damage caused by the different optical isomers on Sertoli cells was in the order of (+) < (+/-) < (-) gossypol. These changes coincided with changes in Sertoli cells of rats following oral administration of gossypol. PMID- 8242830 TI - Characterization of liposomes and other lipid assemblies by multiprobe fluorescence polarization. AB - This minireview describes the use of fluorescence polarization to characterize the dynamics and organization of biological membranes, lipoproteins, liposomes and other lipid-based assemblies. The advantages, disadvantages and limitations of this method are discussed. There is special emphasis on the multiprobe approach, which can monitor various parts of the lipid membrane or assembly. The fluorescence polarization, anisotropy and the total fluorescence intensity are defined and their graphic presentation described. A user guide for the actual measurements is given. It describes how to apply fluorescence polarization, especially how to select the proper extrinsic probe and to optimize the measurement regarding theoretical and practical issues, the determination of a fluorescent probe location in the assembly, how to introduce the probe to the lipid assembly, what is the ideal ratio of the probe to the lipid, dealing with the spectrofluorimeter problems and how to overcome disturbances introduced by light scattering. It is shown here, that even though there are disadvantages in using fluorescence polarization, this approach provides a highly sensitive and simple means for assessing the subtle and more pronounced changes in the structure of lipid assemblies. PMID- 8242831 TI - Structural and motional properties of vesicles as revealed by nuclear magnetic resonance. PMID- 8242832 TI - Osmotic behaviour and permeability properties of liposomes. AB - In this overview liposomes are described as bilayer-bounded vesicles which under defined conditions act as ideal osmometers according to the Boyle van't Hoff law. Investigations on osmotic volume changes, directly or indirectly by taking advantage of changes in light scattering, are considered and applications in permeability measurements are discussed. Solute-solvent interactions occurring in isotonic swelling experiments are analysed in view of an irreversible thermodynamic description. In a second part liquid crystalline lipid bilayers are characterized as highly selective permeability bilayers and the physical principles underlying this selectivity are considered. Attention is given to special physical and chemical conditions that may cause structural defects in the bilayer organization and can affect the selective permeability properties of the bilayer or completely deteriorate its barrier function. Finally an evaluation is given of intrinsic ionophoretic activity in lipid bilayers containing negatively charged lipids. PMID- 8242833 TI - Quality control of liposomal lipids with special emphasis on peroxidation of phospholipids and cholesterol. AB - The usefulness of various assays for the determination of phospholipid and cholesterol peroxidation in liposome formulations was studied on model liposomes prepared as small unilamellar vesicles (SUV) and multilamellar vesicles (MLV) from either native egg phosphatidylcholine (EPC), partially hydrogenated egg phosphatidylcholine (PHEPC) or fully hydrogenated egg phosphatidylcholine (HEPC) and cholesterol in 65/35 molar ratio at a total lipid concentration of 10 mumol/ml in phosphate buffered saline pH 7.2. Liposomes were incubated at 50 degrees C for a total of 3 months. Fatty acid and cholesterol peroxidation were monitored after 1, 2 and 3 months by quantitative measurement of fatty acids and cholesterol and as well as peroxidation products. Fatty acid peroxidation products malondialdehyde, lipidhydroperoxides, conjugated dienes, conjugated trienes were poor predictors of actual fatty acid loss. Among the cholesterol peroxidation products 7-hydroxy-cholesterols, 7-keto-cholesterol and 4-cholesten 3-one were measured quantitatively. Only the formation of 7-keto-cholesterol correlated well with cholesterol disappearance. PMID- 8242834 TI - The determination of liposome captured volume. AB - Manipulating the process by which lipids assemble to form bilayer membranes has produced a myriad of protocol-dependent liposome types. For each of these systems the arrangement of bilayers is characteristic and can be described by parameters such as aqueous entrapment per mole lipid or captured volume, vesicle size distribution, the average number of lamellae per vesicle and shape. For specific applications as model systems or drug delivery systems specific characteristics are desired. Consequently over the years many techniques have evolved to better quantitate these parameters. Here we focus on and detail several methods to quantitate liposome captured volume. We also briefly describe the available methods to measure the other aforementioned physical properties and discuss their interdependency with captured volume. PMID- 8242835 TI - In vitro tests to predict in vivo performance of liposomal dosage forms. AB - Design of liposome-based formulations for clinical use can be assisted by employing in vitro assays to predict pharmacokinetics and bioavailability of the drug before employing costly and time-consuming in vivo studies. For such formulations of the anti-cancer drug doxorubicin (DXR) we developed two assays. (A) An assay which determines the dilution-induced DXR release in buffers and plasma. This assay was employed to evaluate two liposomal DXR formulations: (i) membrane-associated liposomal doxorubicin (L-DXR), and (ii) sterically-stabilized liposomes which encapsulate DXR in the aqueous phase of the liposomes (S-DXR). The agreement between the dilution-induced release assay in vitro and the pharmacokinetics of DXR administrated either as L-DXR or as S-DXR in humans suggests that the dilution release assay can be used as a predictor for the pharmacokinetic performance of liposomal formulations. (B) An assay which determines intracellular drug release induced by liposome degradation in the presence of mouse liver lysosome lysate. This assay was used to assess bioavailability of DXR when delivered via L-DXR, which are taken up by the reticuloendothelial system (RES). (C) An assay which complements conventional chromatographic analyses (HPLC or TLC) of the drug, in which a DXR adduct or aggregate was determined by using Sephadex LH-20 gel exclusion chromatography. PMID- 8242836 TI - Complement-dependent phagocytosis of liposomes. AB - In this article we describe an in vitro model for complement-dependent phagocytosis of liposomes. We have previously reported that complement-opsonized liposomes are avidly ingested by murine peritoneal or bone marrow-derived cultured macrophages. However, when the liposomes contained certain lipids, including phosphatidylinositol, ganglioside GM1, and sulfogalactosyl ceramide, that have been identified as causing prolonged circulation time in vivo, complement-dependent phagocytosis of the liposomes was greatly suppressed. We identify certain additional factors associated with suppressed complement dependent phagocytosis, including, liposomal negative charge and liposomal prostaglandin E2 or thromboxane B2. Possible mechanisms responsible for suppression of complement dependent phagocytosis are suggested. We propose that suppression of complement-dependent phagocytosis could be a contributing factor in the promotion of increased circulation time of 'stealth' liposomes and that complement opsonization probably plays a role in vivo in removing liposomes from the circulation. PMID- 8242837 TI - Surface-modified liposomes: assessment and characterization for increased stability and prolonged blood circulation. AB - Advances in therapeutic applications of liposomes have been achieved through surface modifications increasing their biological stability: reduced constituent exchange and leakage as well as reduced unwanted uptake by cells of the mononuclear phagocytic system. The recent conclusions obtained from in vivo and in vitro studies are reviewed with an emphasis on evaluating the methods used and thus the kinds of conclusions which can be drawn. A number of issues are raised as to the limitations of the methods employed. Steric stabilization, meaning reduction in particle interactions by a surface steric barrier, has been proposed as a theoretical basis for the results and some of the initial results testing this hypothesis are reviewed here with respect to identification of the extent to which physical properties of the surface coatings correlate with the biological properties. At this time it seems that no one method is ideal so that multiple measures give the best characterization. PMID- 8242838 TI - Toxicity screening of liposomes. AB - Phospholipids are the major components of most liposomes. Extensive testing of these naturally occurring compounds has revealed them to be remarkably safe for pharmaceutical use. Addition of other constituents to liposomes in order to alter stability or kinetics can result in an increase in toxic potential, particularly on parenteral administration of liposomes. This paper describes some simple in vitro cellular tests for direct toxicity of liposomes, particularly following intravenous (i.v.) or topical administration, including tests for haemolysis, thrombosis and cytotoxicity. In addition, an in vivo test for the effects on phagocytosis and for pyrogenicity are described, together with a brief outline of the requirements for the further toxicity testing of liposomal drugs at a later stage of development. PMID- 8242839 TI - Liposomes: the Babraham connection. AB - A chronological history is given of the experimental work carried out at The Institute of Animal Physiology, Babraham, Near Cambridge (UK) prior to and after the first description of liposomes in 1965. A convergence of worldwide interest in membrane structure and function coupled with the coexistence of physiologists, biochemists and electron microscopists provided a unique opportunity for the development of a useful model system. As well as defining their properties, such as composition, dimensions, perm-selectivity, permeating pathways and response to solutes (oil and/or water soluble), temperature and pressure, attention is drawn to a number of ideas investigated over the years relating to in vivo 'invisibility' of foreign particles, the trapping of ions in small vesicles the origins of life, to the mechanisms of anaesthesia and formulation of an effective lung surfactant. PMID- 8242840 TI - Chemical stability of liposomes: implications for their physical stability. AB - In the first part of this article, chemical and physical stability of aqueous liposome dispersions have been addressed. Chemical stability of phospholipids has been considered in two parts: oxidation and hydrolysis. Major attention has been paid to hydrolysis kinetics of phospholipids as a function of pH, temperature, buffer concentration and ionic strength. Furthermore, the effect of chain length, head group, state of aggregation, addition of cholesterol and presence of charge on the hydrolysis kinetics of phospholipids has been dealt with. In the second part physical stability of chemically degraded liposome dispersions has been evaluated. In the final part quality control assays for liposome dispersions is presented and a HPLC method with a refractive index detector for the analysis of phospholipids from aqueous liposome dispersions is described. PMID- 8242841 TI - Liposome stability and formation: experimental parameters and theories on the size distribution. AB - The most important characteristics of liposomes, in addition to chemical composition and surface properties, are size distribution and lamellarity. Liposomes can be formed by many different preparation techniques, which according to the literature yield rather well defined vesicle preparations. In contrast to abundant information on the experimental procedures and preparation protocols the theoretical understanding of these processes is lacking. Only geometrical models of structural changes exist for few preparation procedures and size of the liposomes prepared by sonication and detergent depletion method were estimated using simple models. In this paper we first outline different theories on the stability of liposomes and their influence on the size distribution. In the experimental section we shall briefly present the importance of size distribution in experimental work and influence of various experimental parameters on the size distributions obtained. PMID- 8242842 TI - Small angle X-ray scattering: possibilities and limitations in characterization of vesicles. AB - The use of small angle X-ray scattering (SAXS) for characterization of lipid vesicle dispersions is described. The effect of curvature of the membrane, the presence of proteins in the core and on the surface of the membrane, variations in membrane thickness and distribution in the number of bilayers of the vesicles in the dispersion on the scattering curve is discussed. Concerning unilamellar vesicles, either the membrane curvature of vesicles smaller than 50 nm or variations in membrane thickness result in a disappearance of the first node in the scattering curve, even if the bilayer is symmetric with respect to the electron density distribution. In the case of dispersion in which unilamellar as well as multilamellar vesicles are present it is shown that a small fraction of multilamellar liposomes changes the scattering curve dramatically. Liposomes were prepared from various compositions of dipalmitoylphosphatidylcholine (DPPC) and cholesterol hemisuccinate (CHEMS) by the film method. The electron density profile of the bilayers and distribution in the number of bilayers of the liposome dispersions were determined. The average number of bilayers increased as a function of the decrease in CHEMS content. Liposomes with higher CHEMS content than 10 mol% were unilamellar. It seems that increase in charge intercalated in the bilayers resulted in unilamellar vesicles. PMID- 8242843 TI - Use of fluorescent probes to monitor molecular order and motions within liposome bilayers. AB - This article reviews the use of fluorescent probes to monitor the order and dynamics within the acyl chain region of liposome lipid bilayers. Fluorescence anisotropy is first defined and the theoretical framework that allows interpretation of steady-state or dynamic measurements in terms of molecular details is reviewed. The general advantages and/or limitations of fluorescent versus other methods of monitoring membrane order and dynamics are discussed. The properties of two classes of fluorescence probes are then described. The linear probes 1,6-diphenyl-1,3,5-hexatriene (DPH) and parinaric acid (PA) and their derivatives are seen as particularly useful when quantitative interpretation of observations in terms of details of bilayer dynamics and order are critical. Of these, DPH is the more widely and easily used, although parinaric acid has advantages for certain applications. The non-linear probes considered include the anthroyloxyl fatty acids and the recently introduced fluorenyl fatty acid probes. While the geometry and electronic configurations of these probes do not allow for detailed molecular interpretations, these probes can provide unique qualitative information about the state of the lipid bilayer at various positions along the acyl chains. PMID- 8242844 TI - Excretion of food-derived heterocyclic amine carcinogens into breast milk of lactating rats and formation of DNA adducts in the newborn. AB - The distribution, DNA adduction and excretion into breast milk of 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) were examined in lactating female F344 rats with 5 day old pups. Six hours after a single dose (10 mg/kg, p.o.) of radiolabeled IQ, MeIQx or PhIP to lactating dams, radioactivity in the dams was highest in the liver and kidney followed, in descending order, by the mammary gland, omental fat and brain. By 24 h after carcinogen administration, all tissues of the dams showed significantly reduced levels of radioactivity except for omental fat which changed only marginally from 6 to 24 h. 32P-Postlabeling analysis showed that the level of DNA adducts in mammary gland 6 h after dosing was 2.2, 0.7 and 0.2 adducts/10(7) nucleotides for PhIP, IQ and MeIQx respectively. In contrast, in hepatic DNA, the levels of IQ DNA adducts (5.5 adducts/10(7) nucleotides) were 11-fold higher than those of PhIP or MeIQx. The stomach contents, liver, kidney and urine of pups nursed by dams given radiolabeled IQ, MeIQx or PhIP were radioactive, indicating that these carcinogens (and/or metabolites) were excreted into breast milk and absorbed by the pups. After a 6 h suckling period, the amount of PhIP-derived radioactivity in the stomach contents of the pups was approximately 10-fold higher than that seen with IQ or MeIQx. Urine from pups from the three groups was mutagenic in the Ames assay with Salmonella TA98 in the presence of an S9 activating system. IQ-, MeIQx- and PhIP-DNA adducts, at levels in the range of 0.25-0.46 adducts per 10(8) nucleotides, were detected in the livers of pups using the 32P-postlabeling method under intensification conditions. The results from this study indicate that breast milk is a route of exposure of the newborn to heterocyclic amines. The presence of DNA adducts in the tissues of pups further suggests that this route of exposure may have a carcinogenic consequence to the newborn. PMID- 8242845 TI - Diet and carcinogenesis. AB - In summary there is a wealth of information on dietary and nutritional effects on carcinogenesis in laboratory rodents. Experimental studies based on epidemiological evidence, earlier experimental studies and known or predicted cellular, biochemical and molecular effects of nutrients have produced clear evidence that carcinogenesis in laboratory rodents is influenced by dietary intake of calories, fat, lipotropes (choline, methionine), vitamin A and related retinoids, Se, calcium, zinc, fiber, ethanol and a large number of non-nutrient components of foods. For these substances or groups of substances mechanistic hypotheses supported by experimental data and are leading to further research. The information provided will contribute to understanding of basic processes in carcinogenesis as well as of the specific interactions studied, and should contribute to significant advances in preventive medicine. Restriction of caloric intake of rodents by amounts > 10% over a significant portion of their lifetime reduces tumorigenesis. That level of restriction reduces the rate of growth and maturation, and most experiments in this area employ greater restrictions that virtually abolish growth from a young age. Therefore, the observations are of interest in mechanistic studies, but their applicability to preventive medicine requires better definition of the degree and duration of restriction required for a significant effect and the age at which it must be imposed. Restriction of total fat intake and modifications to increase the intake of omega-3 fats have a reasonably consistent effect on tumorigenesis in rodents but a much less consistently demonstrable effect in humans. Again, the observations in rodents are providing a major stimulus to mechanistic studies. The lipotropes are extremely valuable as tools for investigating mechanisms of carcinogenesis in rodents. Their importance in the epidemiology of human cancer has yet to be demonstrated clearly and is a subject of research at present. The naturally occurring vitamins and minerals, as well as fiber, derive their importance in this context from investigations to explain the consistent epidemiological demonstrations of reduction of tumor risk with increased consumption of fruits and vegetables. The activity of the isolated nutrients as anticarcinogens in rodents has generally not matched the activity expected from epidemiological studies. The anticarcinogenic activity of many of the non-nutrient components of fruits and vegetables is remarkable in particular models, however, as is the activity of natural and synthetic retinoids. At present the results must be interpreted to indicate an important effect of combinations of the whole foods with identification of particular nutrients or non-nutrients in specific cases.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8242846 TI - Inhibition by dietary curcumin of azoxymethane-induced ornithine decarboxylase, tyrosine protein kinase, arachidonic acid metabolism and aberrant crypt foci formation in the rat colon. AB - The present study was designed to investigate the modulatory role of dietary curcumin on (i) azoxymethane (AOM)-induced ornithine decarboxylase (ODC), tyrosine protein kinase (TPK) and arachidonic acid metabolism in liver and colonic mucosa of male F344 rats, (ii) in vitro arachidonic acid metabolism in the liver and colonic mucosa and (iii) AOM-induced aberrant crypt foci (ACF) formation in the colon of F344 rats. At 5 weeks of age groups of animals were fed one of the experimental diets containing 0 or 2000 p.p.m. curcumin. Two weeks later all the animals except the vehicle-treated groups were given s.c. injections of AOM, 15 mg/kg body wt, once weekly for 2 weeks. The animals intended for biochemical study were killed 5 days later and the colonic mucosa and liver were analyzed for ODC, TPK, lipoxygenase and cyclo-oxygenase metabolites. The animals intended for ACF study were killed 9 weeks later and analyzed for ACF in the colon. The results indicated that in saline-treated animals dietary curcumin significantly inhibited the ODC (P < 0.001) and TPK (P < 0.05) activities in the liver and colonic mucosa. Dietary curcumin significantly decreased the levels of AOM-induced ODC activity in the liver and colon (P < 0.0001) and TPK activity in the liver and colon (P < 0.01-0.0001) and the formation of 5(S)-, 8(S)-, 12(S)- and 15(S)-hydroxyeicosatetraenoic acids (HETEs) in the liver and colon (P < 0.0001). Also, curcumin suppressed AOM-induced prostaglandin (PG) and thromboxane (Tx) formation in the liver (PGE2, PGF2 alpha, PGD2, 6-keto-PGF1 alpha and TxB2 to 40, 59, 55, 53 and 39% respectively) and in the colon (PGE2 and PGF2 alpha to 39 and 41% respectively). Further, dietary curcumin reduced the in vitro formation of HETEs, PGs and Tx in a dose-dependent manner. AOM-induced colonic ACF were significantly (P < 0.001) inhibited in the animals fed the curcumin diet. The results of the present study indicate that curcumin, present in turmeric, inhibits AOM-induced colonic preneoplastic lesions and other cellular events relevant to colon carcinogenesis. PMID- 8242847 TI - Tumor promotion by an anticonvulsant agent, phenytoin, in mouse liver: correlation with CYP2B induction. AB - To investigate the liver tumor promoting effects of phenytoin (5,5 diphenylhydantoin; DPH), 5 week old male D2B6F1 mice were given a single i.p. dose of 90 mg N-nitrosodiethylamine (NDEA)/kg body wt in tricaprylin. Control groups received tricaprylin alone. After 2 weeks, the mice were given a diet containing 500, 250 or 125 p.p.m. DPH. Ten mice from each treatment group were killed at 30 weeks of age, at which time 10/10 mice given 500 p.p.m. DPH after NDEA initiation had developed multiple hepatocellular foci and adenomas. Such lesions were found only in 2/10 mice given NDEA alone. By 60 weeks, when the experiment was concluded, the incidences (and multiplicities, in units of tumors per tumor-bearing mouse) of hepatocellular adenomas were 60% (1.8 +/- 0.8), 100% (11.6 +/- 5.5), 77% (4.4 +/- 3.3) or 71% (2.6 +/- 1.3) in mice exposed to NDEA alone, or NDEA followed by 500, 250 or 125 p.p.m. DPH respectively. Hepatocellular carcinomas (87% incidence) and hepatoblastomas (33% incidence) were found only in mice given 500 p.p.m. DPH following NDEA initiation. Dose dependent and profound increases in hepatic CYP2B-mediated benzyloxyresorufin O dealkylase activity were detected in livers of B6C3F1 mice exposed for 14 days to dietary DPH (125, 250, 500 or 1000 p.p.m.). Similar increases in this activity were observed in D2B6F1 mice exposed to 500 and 250 p.p.m. for 30 or 60 weeks. Thus, increased hepatic CYP2B activity in mice exposed to DPH correlates with the tumor promoting effect of this compound. PMID- 8242848 TI - Bacterial and mammalian cell mutagenicity of four optically active bay-region 10,11-diol-8,9-epoxides of the nitrogen heterocycle dibenz[a,h]acridine. AB - The mutagenic activities of the enantiomers of the diastereomeric pair of bay region 10,11-diol-8,9-epoxides of dibenz[a,h]acridine (DB[a,h]ACR) were evaluated in histidine-dependent strains of Salmonella typhimurium and in cultured Chinese hamster V79 cells. In strains TA98 and TA100 of S.typhimurium, the (-) [8S,9R,10R,11S] diol-epoxide was the most mutagenic compound, inducing 1200 and 6900 His+ revertants/nmol respectively. The mutagenic activity of each of the remaining three isomers was essentially independent of the bacterial strain used and had 14-72% of the activity of the [S,R,R,S] isomer. However, in Chinese hamster V79 cells, the (+)-[8R,9S,10S,11R] diol-epoxide was the most mutagenic compound (68 8-azaguanine resistant variants/nmol/10(5) cells), inducing from 2 to 11 times as many mutations as the other three isomers. These results are analogous to previous studies with the bay-region diol-epoxides of other polycyclic hydrocarbons in that the isomer with [R,S,S,R] absolute configuration has had variable activity in the bacterial assays, but has generally been the most active in the mammalian cells. Furthermore, this isomer has almost always been highly tumorigenic in the mouse. PMID- 8242849 TI - Increases in 13-hydroxyoctadecadienoic acid dehydrogenase activity during differentiation of cultured cells. AB - Recently, oxidation products of linoleic acid such as 13-hydroxyoctadecadienoic acid (HODE) have been implicated in the regulation of cellular physiology including the proliferative response to growth factor treatment. In addition, an NAD(+)-dependent 13-HODE dehydrogenase was recently described. To evaluate the contribution of this enzyme to cellular processes we have examined the behavior of the enzyme under different conditions. In the present report, changes in the activity of 13-hydroxyoctadecadienoic acid dehydrogenase during in vitro differentiation of two different cell lines were examined. The cell line HT-29 undergoes induced differentiation via manipulation of the medium while the Caco-2 line undergoes spontaneous differentiation upon attainment of confluence. In both cell lines, longer culture times were accompanied by increases in 13-HODE dehydrogenase activity. The increase in enzyme activity continued even after cell proliferation had ceased. Cellular differentiation was verified by the observation of increases in sucrase and alkaline phosphatase activities. In addition, the activity of 13-HODE dehydrogenase was measured in growing, early confluent and late confluent cultures of undifferentiating Swiss mouse 3T3 fibroblasts. In the fibroblast line, no significant changes in 13-HODE dehydrogenase activity were observed during the course of the experiment. The specific activity of 13-HODE dehydrogenase was also significantly different between the three cell lines, consistent with the extent of differentiation. Highest levels of activity were found in Caco-2 cells (200-400 pmol/min/mg) and barely detectable levels in the fibroblasts (0.6-2 pmol/min/mg). The correlation between 13-HODE dehydrogenase and cell differentiation suggests the enzyme may have a role to play in the partitioning of cells between proliferation and differentiation pathways. PMID- 8242850 TI - N-acetoxy-N-acetyl-2-aminofluorene-induced mutation spectrum in a human hprt cDNA shuttle vector integrated into mammalian cells. AB - The spectrum of mutations induced by N-acetoxy-N-acetyl-2-aminofluorene (N-AcO AAF) was examined by the pZipHprtNeo shuttle vector in mammalian cells. The vector carries a cDNA of the human hypoxanthine phosphoribosyl transferase (hprt) gene, which is stably integrated into chromosomal DNA of a mouse cell line, VH12. After treatment of the cell with N-AcO-AAF, 48 independent 6-thioguanine resistant clones were obtained and altered sequences of the mutated cDNA hprt genes were determined. Frameshifts and deletions were the predominant mutational events (68%) induced by N-AcO-AAF and the remainder were base substitutions (32%) of various types. Analysis of sequence alterations at all the sites of mutation revealed that: (i) > 65% of mutations occurred at G:C sites, suggesting C8G adducts are responsible premutagenic lesions for these mutations; and (ii) short sequence repeats were frequently found at the sites of frameshift and deletion, and slippage--misalignment is the suggested mechanism for the induction of mutations at these sites. Implied significance of slippage--misalignment as a fundamental mechanism for mutagenesis is discussed. PMID- 8242851 TI - DNA strand breakage and DNA adducts in lymphocytes of oral cancer patients. AB - In lymphocytes of 12 oral cancer patients (and two control groups) the frequencies of DNA single-strand breaks and DNA-protein cross-linking were determined by alkaline filter elution. We found elevated DNA elution rates, which must be interpreted as an increased strand breakage frequency. There were significant correlations between the DNA strand breakage frequency and smoking habits. Using the 32P-postlabelling assay we determined the DNA adduct level in lymphocytes of 23 oral cancer patients (and two control groups). No significant influence of smoking habit on the DNA adduct level could be detected. There was a significant correlation between the DNA adduct level and the gamma glutamyltranspeptidase (GGT) value, suggesting systemic influences of alcohol drinking habits on the adduct level. PMID- 8242852 TI - Phosphatase inhibitors, gap junctional intercellular communication and [125I]-EGF binding in hamster fibroblasts. AB - A number of phosphatase inhibitors (okadaic acid, calyculin A, aluminium fluoride, sodium molybdate, sodium orthovanadate, pervanadate and vanadyl sulphate) were investigated for their effects on gap junctional intercellular communication (GJIC) and [125I]-epidermal growth factor (EGF) binding in early passage Syrian hamster embryo cells (mainly fibroblast-like cells) and in V79 Chinese hamster lung fibroblasts. Only pervanadate decreased GJIC significantly. After the initial pervanadate-induced decrease the GJIC recovered rapidly. Only pervanadate was able to change the band pattern of the gap junction protein connexin43 (cx43) in Western blots. Together this may indicate either that there is a low turnover of phosphate groups in cx43 under basal conditions or that the putative phosphatases are not sensitive to most of the phosphatase inhibitors applied. In contrast, pervanadate, orthovanadate and molybdate decreased [125I] EGF binding. 12-O-Tetradecanoylphorbol-13-acetate (TPA) is able to induce the phosphorylation of both cx43 and the EGF receptor, concomitantly with a decrease in GJIC and [125I]-EGF binding. These effects are reversible after removal of TPA. It could be imagined that other phosphatases would act on cx43 and the EGF receptor after the forced phosphorylation of the two molecules. Thus TPA was used to downregulate GJIC and [125I]-EGF binding and phosphatase inhibitors were applied in the upregulation phase. Only pervanadate affected the upregulation of GJIC, and pervanadate, orthovanadate and molybdate affected the upregulation of [125I]-EGF binding. Thus it is not an identical complement of phosphatases that act on cx43 and the EGF receptor. All the downregulating agents are assumed to be phosphotyrosine phosphatase inhibitors. PMID- 8242853 TI - Development of hydroxysteroid sulfotransferase-deficient lesions during hepatocarcinogenesis in rats. AB - Rat liver cytosolic hydroxysteroid sulfotransferases form highly reactive sulfuric acid esters from some benzylic alcohols, such as 1-hydroxymethylpyrene. In this study we examined the expression of hydroxysteroid sulfotransferase a (STa) in carcinogen-induced enzyme-altered, presumably preneoplastic, rat liver foci. Female Wistar rats were given a single i.p. injection of diethylnitrosamine (0.15 mumol/g body wt) 1 day after birth to induce the liver foci. After weaning, rats were given 1-hydroxymethylpyrene or phenobarbital continuously in their diet (250 or 500 p.p.m. respectively) for a total of 120 days. Carcinogen-induced liver foci were identified by a change in the marker enzyme adenosine triphosphatase. Immunohistochemical staining of consecutive sections using an anti-STa rabbit antibody demonstrated that STa was expressed at decreased levels in most of the adenosine triphosphatase-negative liver foci. This effect was observed in both 1-hydroxymethylpyrene- and phenobarbital-treated animals. The decrease in STa content in enzyme-altered foci may lead to a selective advantage of the preneoplastic cells in the presence of agents that are able to form reactive sulfuric acid esters, such as 1-hydroxymethylpyrene. In some diethylnitrosamine/phenobarbital-treated rats, a small number of atypical foci were observed, most of them showing enhanced expression of STa and unchanged to moderately increased ATPase activity. PMID- 8242854 TI - DNA damage induced by furocoumarin hydroperoxides plus UV (360 nm). AB - When irradiated at 360 nm, furocoumarins with a hydroperoxide group in a side chain efficiently give rise to a type of DNA damage that can best be explained by a photo-induced generation of hydroxyl radicals from the excited photosensitizers. The observed DNA damage profiles, i.e. the ratios of single strand breaks, sites of base loss (AP sites) and base modifications sensitive to formamidopyrimidine--DNA glycosylase (FPG protein) and endonuclease III, are similar to the DNA damage profile produced by hydroxyl radicals generated by ionizing radiation or by xanthine and xanthine oxidase in the presence of Fe(III) -EDTA. No such damage is observed with the corresponding furocoumarin alcohols or in the absence of near-UV radiation. The damage caused by the photo-excited hydroperoxides is not influenced by superoxide dismutase (SOD) or catalase or by D2O as solvent. The presence of t-butanol, however, reduces both the formation of single-strand breaks and of base modifications sensitive to FPG protein. The cytotoxicity caused by one of the hydroperoxides in L5178Y mouse lymphoma cells is found to be dependent on the near-UV irradiation and to be much higher than that of the corresponding alcohol. Therefore the new type of photo-induced damage occurs inside cells. Intercalating photosensitizers with an attached hydroperoxide group might represent a novel and versatile class of DNA damaging agents, e.g. for phototherapy. PMID- 8242855 TI - Severity of promotion by sodium salts of succinic acid in rat urinary bladder carcinogenesis correlates with sodium ion concentration under conditions of equal urinary pH. AB - Elevation of urinary pH and Na ion concentration induced by feeding high doses of sodium salts plays a crucial role in promotion of urinary bladder carcinogenesis in male rats initially treated with N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN). In the present study, urinary bladder carcinogenesis in male F344 rats was promoted by feeding the mono- and di-sodium salts of succinic acid, and the activity was enhanced by varying urinary sodium ion concentration under conditions of equally high pH. In Experiment 1, the rats were given 0.05% BBN in their drinking water for 4 weeks and then fed basal diet containing 5% succinic acid, 5% sodium succinate (Na-Suc) or 5% di-sodium succinate (2Na-Suc) for 32 weeks. The development of carcinomas in the urinary bladder was enhanced by treatment with Na-Suc and with 2Na-Suc. 2Na-Suc induced larger urinary bladder tumors than the Na-Suc. In Experiment 2, 5% Na-Suc and/or 5% 2Na-Suc were fed to the rats for 8 weeks without prior BBN treatment. Induction of simple hyperplasia as observed by light microscopy, was greater in rats fed 2Na-Suc than Na-Suc. Increased 5-bromo-2'-deoxyuridine labeling index and alterations of the urothelial surface observed by scanning electron microscopy of the urinary bladder were similarly greater in rats fed 2Na-Suc compared to Na-Suc. In addition, there was a tendency toward increased spermidine/spermine N1 acetyltransferase activity in the urinary bladder epithelium of rats fed 5% 2Na Suc. The results of Experiment 2 corresponded to differences in promoting activity for the different chemicals in Experiment 1. Thus, tumor growth was associated with sodium ion concentration level under conditions of equal increase of urinary pH. PMID- 8242856 TI - Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on protein kinase C and inositol phosphate metabolism in primary cultures of rat hepatocytes. AB - Adult rat hepatocytes, after maintenance for 24 h in serum-free culture, were treated with the tumor promoters, 12-O-tetradecanoylphorbol-13-acetate (TPA) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Short-term treatment (15 min) with TPA, 1 microM, increased protein kinase C (PKC) activity in the particulate fraction of hepatocytes and, concomitantly, decreased the vasopressin (100 nM) stimulated synthesis of inositol phosphates. The latter effect of TPA could be prevented by prior addition of the PKC inhibitor, H7 (100 microM). After short term treatment (15 min) with TCDD, 1 pM, no effects on PKC or inositol phosphate metabolism were observed. However, after prolonged exposure to TCDD (3-48 h), the particulate PKC was significantly activated (1.5-fold). In contrast to the effect of TPA (24 h), no down-regulation was found. Moreover, long-term treatment with TCDD significantly enhanced vasopressin-stimulated inositol 1,3,4,5 tetrakisphosphate synthesis, while TPA treatment (24 h) stimulated the synthesis of inositol trisphosphates and inositol 1,3,4,5-tetrakisphosphate. The results suggest that the tumor promoters, TPA and TCDD, act differently on the signal transduction pathways in hepatocytes. Thus, the effects of TCDD on PKC and inositol phosphate metabolism might be mediated by a yet unknown mechanism rather than by direct activation of PKC as seen with TPA. PMID- 8242857 TI - Characterization of benzo[a]pyrene-initiated mouse skin papillomas for Ha-ras mutations and protein kinase C levels. AB - The frequency and spectrum of Ha-ras mutations in benzo[a]pyrene (B[a]P) initiated/12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted CD-1 mouse skin papillomas were characterized by amplifying high molecular weight papilloma DNA using the polymerase chain reaction (PCR) followed by direct DNA sequencing. Analysis of 10 individual B[a]P-initiated early emergence papillomas indicated that 90% contained a Ha-ras mutation. Twenty percent of these papillomas contained a GGA-->GTA transversion in the 12th codon, 50% contained a GGC-->GTC transversion in the 13th codon and 20% contained a CAA-->CTA transversion in the 61st codon. A characteristic of 7,12-dimethylbenz[a]anthracene (DMBA)-initiated papillomas, which contain an A-->T mutation in the 61st codon of Ha-ras, is that they exhibit a constitutive decrease in both protein kinase C (PKC) activity and PKC alpha and beta 2 isozyme levels when compared to epidermis. In the present study we found that total PKC activity, as well as PKC alpha and beta 2 isoforms, were markedly decreased in B[a]P-initiated early emergence papillomas and that this decrease was also accompanied by an altered subcellular distribution of PKC activity. The particulate/cytosolic (P/C) ratio of PKC activity in the epidermis was 0.39, whereas the P/C ratio in the papillomas was 0.77. These results demonstrate that B[a]P-initiated/TPA-promoted papillomas exhibit a high incidence of specific ras mutations and that PKC levels are constitutively decreased in these papillomas, indicating that an activated ras gene is associated with and may contribute to the observed decrease in PKC levels. PMID- 8242858 TI - Role of SV40 T antigen binding to pRB and p53 in multistep transformation in vitro of human uroepithelial cells. AB - In the present study, we examined the sufficiency of SV40 T antigen (Tag) binding to pRB and p53 to substitute for alterations in RB and TP53 at all stages of human uroepithelial cell (HUC) transformation in vitro. Two independent SV40 immortalized HUCs (SV-HUC and SV-HUC/CK2) and 17 independent derivative carcinogen-induced or spontaneous tumors (T-SV-HUCs and T-SV-HUC/CK2) representing different stages of urothelial tumorigenesis were examined. Although five of 17 T-SV-HUCs and SV-HUC/CK2 and its derivative tumor showed 13q chromosome deletion and loss of heterozygosity (LOH), this did not reflect functional loss of pRB because Tag/pRB binding was unaltered and sequencing showed a normal RB gene in all these tumors. No genetic alterations involving 17p or TP53 were detected in any tumors in this study using the same techniques. These results indicate that Tag/pRB and Tag/p53 binding apparently abrogate requirements for/or a selective advantage of RB and TP53 mutations in HUC tumorigenic transformation and progression, as well as in HUC immortalization. These data also provide new evidence that more than one suppressor gene may be located on chromosome 13q. PMID- 8242859 TI - Increased mutagenicity of 1,2-dibromo-3-chloropropane and tris(2,3 dibromopropyl)phosphate in Salmonella TA100 expressing human glutathione S transferases. AB - We have expressed human glutathione S-transferases GSTA1-1 and GSTP1-1 in Salmonella typhimurium TA100 in order to assess the ability of these enzymes to modulate the mutagenicity of 1,2-dibromo-3-chloropropane (DBCP) and tris(2,3 dibromopropyl)phosphate (Tris-BP). Both compounds were mutagenic when activated by Aroclor-induced rat liver microsomes. However, when Aroclor-induced rat liver microsomes were used together with the GST-expressing strains the mutagenicity of both DBCP and Tris-BP was markedly potentiated. Neither of the GST-expressing strains potentiated the mutagenicity in the absence of microsomes, indicating that cytochrome P450-mediated metabolism was a prerequisite for GST-mediated potentiation. With DBCP both isozymes had comparable effects on mutagenic frequency, although the highest dose of DBCP was toxic in strains expressing GSTP1-1. In the case of Tris-BP, GSTP1-1 was much more active in potentiating the mutagenicity. These results indicate that human GSTs can play an important role in the activation of compounds such as DBCP and Tris-BP to mutagenic metabolites. PMID- 8242860 TI - Quantification of oxidative DNA modifications in mitochondria. AB - Specific repair endonucleases were used to quantify oxidative modifications in mitochondrial DNA (mtDNA) from rat liver and from porcine liver and kidney by means of a relaxation assay. In rat liver mitochondria the number of modifications sensitive to formamidopyrimidine--DNA glycosylase (FPG protein), which include 8-hydroxyguanine (8-oxo-7,8-dihydroguanine) residues, was only 0.8 +/- 0.2 per 10(5) base pairs (bp). Even lower values were observed in porcine kidney (0.5 +/- 0.3 per 10(5) bp) and liver (0.4 +/- 0.2 per 10(5) bp). The numbers of sites of base loss (AP sites) sensitive to T4 endonuclease V and of 5,6-dihydropyrimidines sensitive to endonuclease III were less than 0.2 per 10(5) bp in all cases. The data provide evidence that the steady-state levels of oxidative mtDNA modifications are low under physiological conditions, either because reactive oxygen species generated in the mitochondria are instantly inactivated or because of efficient DNA repair processes inside mitochondria. PMID- 8242861 TI - Heterocyclic aromatic amine formation in grilled bacon, beef and fish and in grill scrapings. AB - The heterocyclic aromatic amines (HAAs) 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino 3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx) and 2-amino alpha carboline (A alpha C) were quantitated in grilled bacon and beef. The levels of PhIP in bacon ranged from < 0.1 to 52 p.p.b., MeIQx was detected at levels ranging from 0.9 to 18 p.p.b. Both 4,8-DiMeIQx and A alpha C were found at < 1 p.p.b. In grilled meat patties, MeIQx and PhIP were detected at levels ranging from 0.8 to 3.2 p.p.b., while 4,8-DiMeIQx and A alpha C were below the limit of detection (approximately 0.5 p.p.b.). HAAs were below the limit of detection in grilled fish. The bacon fat drippings and the pan scrapings obtained from grilled meat and fish also contained significant amounts of HAAs and indicated that either these carcinogens are released with the fat during grilling or that HAAs are formed directly in the released fat and juices. Several of these carcinogens were detected in the pan scrapings at concentrations 10- to 100-fold higher than in cooked meats. PhIP was detected at 144 p.p.b. in combined grilled meat and fish scrapings, followed by A alpha C at 77 p.p.b., MeIQx at 29 p.p.b. and 4,8 DiMeIQx at 4 p.p.b. The co-mutagens harman and norharman were also detected in cooked meats and fish at amounts ranging from 5 to 30 p.p.b. Fat drippings and grill residue scrapings are often used as a base for gravies and sauces. Thus, cooking practices and dietary habits have a strong impact on HAA exposure. PMID- 8242862 TI - Induction of micronuclei and karyotype aberrations during in vivo mouse skin carcinogenesis. AB - The induction of chromosome and/or genome mutations during the first steps of skin carcinogenesis was followed in male NMRI mice, treated with a 'two-stage' [9,10-dimethyl-1,2-benzanthracene (DMBA) + phorbol-12-myristate-13-acetate (TPA)], or a 'three-stage' [DMBA+methyl methanesulphonate (MMS) + phorbol-12 retinoate-13-acetate (RPA)] protocol. The scoring of micronuclei (MN) in basal and suprabasal keratinocytes allows a relatively fast in vivo estimation of clastogenic and aneugenic effects of various compounds and treatments. Relevant stages were then further analysed by karyotyping the in vivo treated keratinocytes that were allowed to divide during short in vitro cultivation. DMBA used as initiator in both protocols was able to induce MN. The well-known clastogen MMS had an acute but transient effect on MN induction when used alone or as convertor in the three-stage protocol. Neither the propagator RPA, nor the 'full-promotor' TPA, which can carry out conversion as well as propagation, induced statistically significant numbers of MN when applied on mouse skin. Combined treatments, DMBA+MMS and MMS+RPA, showed higher MN frequencies than when MMS treatments were given alone. The full carcinogenic protocols showed significant frequencies of MN but the time points of appearance differed, indicating that the accumulation of aberrations could be more important than the order of appearance. Karyotypic analysis of those stages where the MN assay detected genome and/or chromosome aberrations revealed no specific loss of chromosomes that might be directly related to the carcinogenic process. When chromosome loss and aberrations were both taken into consideration together, chromosomes 7 and 11 and surely 9, 17 and 18 were more frequently involved than others. PMID- 8242863 TI - Peroxidase activation of hydroquinone results in the formation of DNA adducts in HL-60 cells, mouse bone marrow macrophages and human bone marrow. AB - Metabolism of benzene results in the formation of multiple metabolites, including hydroquinone (HQ). HQ is a reducing co-substrate for peroxidase enzymes, and the resultant semiquinone and para-benzoquinone (p-BQ) may bind to DNA. The role of peroxidase activation in the formation of DNA adducts by benzene metabolites has not been established. In this study we investigated the role of peroxidase activation in the formation of DNA adducts by HQ and p-BQ in HL-60 cells, human bone marrow (HBM) cells, mouse bone marrow macrophages (MBMM) and the U-937 and Raji leukemia cell lines. Adduct formation was measured by P1-enhanced 32P postlabeling; peroxidase activity was measured with a spectrophotometric assay. Treatment with p-BQ resulted in the formation of two DNA adducts in all of the cell lines. The DNA adducts were identical in all of the cells, however, the adduct level varied by 80-fold. Treatment with HQ produced one DNA adduct in HL 60 cells, HBM and MBMM; no adducts were detected in U-937 or Raji cells. The HQ DNA adducts in the three cell lines were identical. The adduct level was highest in the HL-60 cells, followed by HBM and MBMM. There was a statistically significant correlation between peroxidase activity and the formation of HQ-DNA adducts. These results suggest that peroxidase-mediated metabolism is involved in the activation of HQ to form DNA adducts in mouse bone marrow and HBM. PMID- 8242864 TI - Selenium and difluoromethylornithine additively inhibit DMH-induced distal colon tumor formation in rats fed a fiber-free diet. AB - We investigated the effects of difluoromethylornithine, an inhibitor of ornithine decarboxylase (ODC) and selenium supplementation on tumor formation induced by the carcinogen 1,2-dimethylhydrazine (DMH) in Sprague-Dawley rats. A biochemical link between polyamine biosynthesis and selenium metabolism to its cancer preventative form has been suggested by the common requirement of S adenosylmethionine. One-hundred and twenty male Sprague-Dawley rats were divided into experimental (n = 80) and control (n = 40) groups. Experimental animals received DMH 20 mg/kg s.c. for 20 weeks. Animals were fed either a regular diet (selenium content 0.2 p.p.m.) or a high selenium diet (5 p.p.m.) with or without 0.2% DFMO in the drinking water. At death, week 30, animal weights within experimental or control groups were not different between the four diet treatment groups. Tumor number and incidence in the proximal colon was not affected by DFMO treatment, selenium supplementation or the combined treatment. In contrast, in the distal colon, 19 tumors developed in the DFMO treated group, 22 tumors in the high selenium group and only 12 tumors in the combined high selenium/DFMO treatment group compared to 32 tumors in the regular diet group. Similarly, tumor incidence was decreased by DFMO and selenium supplementation and their effects were additive. In control animals, ODC activity was decreased by DFMO treatment and selenium supplementation in the distal colon and liver, but not the proximal colon. ODC activity of tumor tissue was greater than normal colon tissue from diet paired animals for proximal and distal colon, except for distal colonic tumors in the high selenium/DFMO treatment group. Polyamine content, however, did not correlate with ODC activity in normal or neoplastic tissue. In general, S adenosylmethionine levels from normal colon and liver tissue were unaffected by diet treatment. Selenium supplementation in combination with DFMO treatment selectively inhibited distal colon tumor formation in rats fed a fiber-free diet. PMID- 8242865 TI - Various inhibitors of DNA topoisomerases diminish repair-specific DNA incision in UV-irradiated human fibroblasts. AB - A function for topoisomerases I and II in DNA excision repair can be postulated from the organization of the mammalian chromosome, involving nucleosomal structures and matrix-attached DNA loops. To analyse this function we determined UV-induced DNA incision in confluent human fibroblasts in the presence of 16 inhibitors of topoisomerases I and II which belonged to at least five different drug categories, based on their mechanism of action. Dose-response experiments were performed, analysed by linear regression and the concentrations at which DNA incising activity was reduced to 50% were calculated (K50 values). The majority of these values represent concentrations for which interfering cell toxicity could be excluded. K50 concentrations, which were determined by extrapolating dose-response data, may hit the toxicity range, nevertheless, we deem our K50 scale useful for making biochemical comparisons. With respect to topoisomerase I, camptothecin and topotecan diminished repair-specific DNA incision to a small extent, whereas distamycin, which binds to the minor groove of DNA, caused a stronger effect. With respect to topoisomerase II the results were as follows. (i) The DNA intercalator ethidium bromide decreased DNA-incising activity at rather low concentrations, which indicates marked inhibitory potency. Quinacrine was less effective. (ii) Inhibitors intercalating and binding to the 'cleavable' DNA-topoisomerase complex (m-AMSA, mitoxantrone, doxorubicin and daunorubicin) strongly suppressed reparative DNA incision. (iii) Only small effects were observed using several drugs which act by trapping the 'cleavable' DNA-enzyme complex, namely nalidixic acid and oxolinic acid. In contrast, etoposide and teniposide inhibited post-UV DNA cleavage sizeably. (iv) Merbarone had to be applied at very high concentrations to reduce UV-induced DNA incision. (v) Novobiocin, an inhibitor of the ATPase subunit of topoisomerase II, markedly diminished repair-specific DNA cleavage. A comparison of the K50 values for DNA incision with those for DNA repair synthesis (1) shows that the majority of the investigated drugs inhibited both repair parameters. There were, however, differences in the concentrations required to achieve the 50% inhibition level. The results are best explained by assuming that in UV-irradiated human fibroblasts the 180 kd form of topoisomerase II is a target enzyme for inhibitors which suppressed repair and that this isozyme is involved in steps preceding repair-specific DNA incision. PMID- 8242866 TI - FVB/N mice: an inbred strain sensitive to the chemical induction of squamous cell carcinomas in the skin. AB - The widespread use of FVB/N mice for the establishment of transgenic lines containing active oncogenes suggested the importance of testing the parent FVB/N mice for sensitivity to experimental carcinogenesis. After initiation of mouse skin by a single treatment with 7,12-dimethylbenz[a]anthracene (DMBA) and promotion by 20 weekly applications of 12-O-tetradecanoylphorbol-13-acetate (TPA), the skin tumor incidence was compared in FVB/N mice, TPA-sensitive (SENCAR and CD-1) and TPA-resistant mice (BALB/c and C57BL/6). Initiation by 25 micrograms DMBA followed by promotion with a low dose of TPA (2 micrograms/week) induced one or more papillomas in only 25% of FVB/N mice, compared with 100% in SENCAR, 53% in CD-1, 17% in BALB/c and 0% in C57BL/6 mice. At a more effective dose of TPA (5 micrograms/week), FVB/N mice initiated by 5, 25 or 100 micrograms DMBA developed 3.4, 6.9 and 11.8 papillomas per mouse. In contrast, the incidence of squamous cell carcinomas (SCCs) (17-18/30 mice) did not increase with DMBA dose. TPA promotion of non-initiated mice induced only six papillomas, but three progressed to SCCs, a high rate of malignant conversion. Skin tumor induction by 20 weekly treatments with 10 micrograms DMBA produced few papillomas, but 50.0% of the papillomas progressed to carcinomas in FVB/N mice, compared with 9.15% in SENCAR, 37.5% in CD-1, 23.1% in BALB/c and 15.0% in C57CL/6 mice. The first carcinomas appeared after 14 weeks in FVB/N, 24 weeks in SENCAR, 26 weeks in CD-1 and C57BL/6 and 34 weeks in BALB/c mice. Thus, FVB/N mice develop an unusually high incidence of SCCs after treatment with repeated DMBA, DMBA initiation-TPA promotion and even TPA alone. PMID- 8242867 TI - Modification of carcinogenesis by alpha-tocopherol, t-butylhydroquinone, propyl gallate and butylated hydroxytoluene in a rat multi-organ carcinogenesis model. AB - Effects of the dietary antioxidants alpha-tocopherol (alpha-Toc), t butylhydroquinone (TBHQ), propyl gallate (PG) and butylated hydroxytoluene (BHT) were examined using a multi-organ carcinogenesis model. Groups of 20 F344 male rats were treated with a single intragastric administration of 100 mg/kg body weight N-methyl-N'-nitro-N-nitrosoguanidine, a single intragastric administration of 750 mg/kg body weight N-ethyl-N-hydroxyethylnitrosamine, two subcutaneous injections of 0.5 mg/kg body weight N-methylbenzyl-nitrosamine and four subcutaneous injections of 40 mg/kg body weight 1,2-dimethylhydrazine. At the same time the rats were given 0.1% N-dibutylnitrosamine for 4 weeks and then 0.1% 2,2'-dihydroxy-di-n-propylnitrosamine for 2 weeks in the drinking water, for a total carcinogen exposure period of 6 weeks. Starting 3 days thereafter the rats received 1% alpha-Toc, 1% TBHQ, 1% PG or 0.7% BHT in the diet, or basal diet alone. Further groups of 10-15 animals each were treated with antioxidant alone or basal diet alone as controls. Surviving animals were killed at the end of week 36. Histopathological examination showed that alpha-Toc increased the incidence of glandular stomach atypical foci but reduced the incidence and multiplicity of kidney atypical tubules. TBHQ significantly elevated the incidences of esophageal papillary or nodular (PN) hyperplasias and papillomas, as well as forestomach papillomas, but significantly decreased the multiplicity of colon adenocarcinomas. PG was only effective in reducing the multiplicity of kidney atypical tubules. BHT enhanced the development of thyroid hyperplasias, but strongly reduced the incidence and multiplicity of colon adenocarcinomas. This compound was also associated with lowered incidence and multiplicity of renal cell tumors. None of the agents studied was unequivocal in exerting either positive or negative influence. PMID- 8242868 TI - Cytochrome P450-mediated metabolism of tumour promoters modifies the inhibition of intercellular communication: a modified assay for tumour promotion. AB - The role of metabolism of tumour promoters on the inhibition of intercellular communication was investigated in a modified V79 metabolic cooperation system. V79 cells, which stably express different rat cytochrome P450 enzymes (CYP1A1, CYP1A2 or CYP2B1), were used in the metabolic cooperation assay. The inhibitory effect on intercellular communication of four compounds was changed in cells expressing cytochrome P450 enzymes, compared to cells without. The phorbol ester TPA and di(2-ethylhexyl)phthalate blocked intercellular communication in all the cell lines tested, but expression of CYP1A1 enzyme reduced the inhibitory activity in these cells. Diethylstilbestrol caused inhibition only with cells containing cytochrome P450 enzymes. In contrast, the benzene metabolite hydroquinone inhibited metabolic cooperation preferentially in cells without cytochrome P450 enzymes. The inhibition of metabolic cooperation by another benzene metabolite, phenol, was not affected by the cytochrome P450 enzymes. The inhibitory activity of several chemicals that have not been tested previously was analysed in the new metabolic cooperation assay. The inhibitory activity of none of these chemicals was affected by cytochrome P450-associated metabolism. 7 Octylindolactam V was as potent as TPA, whereas the related indolactam V was 100 fold less active. The carcinogenic aromatic amine 4-aminobiphenyl, but not its primary metabolite 4-hydroxyaminobiphenyl, inhibited metabolic cooperation. Other known carcinogens, ochratoxin A, aflatoxin B1 and 4-nitrobiphenyl, did not inhibit metabolic cooperation in either V79 cells expressing or cells not expressing cytochrome P450. We conclude that cytochrome P450-associated metabolism plays an important role in the inhibition of gap junctional intercellular communication of some tumour promoters. The modified metabolic cooperation assay presented here is valuable for detecting some inhibitory chemicals which have been 'false negative' in previous assays for gap junctional intercellular communication. The assay also discloses that cytochrome P450 metabolism alters intercellular communication by a mechanism other than metabolism of the exogenous inhibitor. PMID- 8242869 TI - Life-span and cancer: the induction time of tumors in diverse animal species treated with nitrosodiethylamine. AB - To ascertain the possible relationship between animal lifespan and the rate of tumor development, the results of carcinogenesis studies in various species treated with similar doses of a carcinogenic nitrosamine have been compiled from the literature. Comparable experiments in 20 species of mammals, reptiles, birds, amphibians and fish were analyzed. The animals received approximately 1000 mg/kg body wt (400-2500 mg/kg) lifetime total dose of nitrosodiethylamine (NDEA). Animals with lifespans varying from 3 years (mouse) to > 50 years (snake) developed tumors with latent periods of roughly 1 year (range 0.5-1.9 year), showing no relationship to lifespan. The evidence suggests that the time dependence of tumor development is more likely related to the cumulative dose of carcinogen than to lifespan and the rate of aging. PMID- 8242870 TI - Metabolism and macromolecular interaction of the tobacco-specific carcinogen 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone in cultured explants and epithelial cells of human buccal mucosa. AB - Metabolism and macromolecular interaction of the tobacco-specific carcinogen 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) were studied in human buccal mucosa in vitro. Microautoradiographic analysis of [5-3H]NNK-exposed explant cultures demonstrated a uniform distribution of bound radioactivity in the mucosal epithelium, without significant binding in the underlying connective tissue. The metabolism of [5-3H]NNK at concentrations of both 6 and 100 microM resulted in seven identified metabolites in both explant and epithelial cell cultures. Formation of 4-(methylnitrosamino)-1-(3-pyridyl)butan-1-ol by carbonyl reduction of NNK accounted for almost 95% of the total metabolism, whereas the proportions of other metabolites obtained by alpha-carbon hydroxylation and pyridine N-oxidation reactions varied from undetectable levels to approximately 2% in both experimental systems. A positive correlation between concentration and the metabolic route associated with the formation of DNA methylating intermediates (alpha-hydroxylation at the N-methylene carbon) was found, i.e. when the concentration of NNK was raised from 6 to 100 microM, keto-acid formation which in part reflects DNA methylation was increased preferentially over ketoalcohol production, an index of DNA pyridyloxobutylation. Both the total rate of NNK metabolism and the amount of protein adducts were higher in cells from primary cultures up to the third passage than in explants cultured for 1 day. Between 10(-9) to 10(-4) M, neither NNK nor its precursor alkaloid nicotine affected the colony forming efficiency of normal and tumorous buccal epithelial cells, although at 10(-3) M each agent inhibited this function. Taken together, the results demonstrate the capability of human buccal mucosal epithelium to metabolize NNK by three major pathways, including those involved in the formation of adducts with cellular macromolecules. PMID- 8242871 TI - Frequency and DNA content of micronuclei in rat parenchymal liver cells during experimental hepatocarcinogenesis. AB - Liver carcinogenesis is considered to be a good experimental model to study the sequential changes leading to cancer and was applied here for the analysis of chromosome/genome mutations. Since the micronucleus test was shown to be an adequate method to detect and analyse chromosome changes in dividing cells, the frequency of micronuclei (MN) together with their relative DNA content (DNA content of the MN divided by the DNA content of the corresponding nucleus) were analysed in hepatocytes isolated from rats at different stages of experimentally induced hepatocarcinogenesis. The protocol used for the induction of liver cancer was based on the triphasic 'Gerlans protocol', a Solt-Farber procedure supplemented with a phenobarbital (PB) promotion step. Male Wistar rats were initiated by a single i.p. dose of diethylnitrosamine (DENA), followed by selection of the resistant hepatocytes by 2-acetylaminofluorene (2-AAF). Subsequent promotion was accomplished by chronic exposure to phenobarbital. For each group of rats a mitotic stimulator (CCl4) is necessary at the end of their treatment period to express the clastogenic and/or aneugenic lesions which may lead to micronuclei. The results of these experiments do confirm that genetic alterations are occurring at the chromosome level (MN expression) during the different steps of experimental rat liver carcinogenesis. DNA measurements seem to be a good genetic parameter to detect eventual differences between the chromosomal content (whole chromosome or chromosome fragments) of MN populations appearing in different stages of the carcinogenic process. Moreover, a comparison between the mono- and bi-nucleated cell population showed that the frequency of micronuclei is higher in mononuclear parenchymal liver cells. PMID- 8242872 TI - Glutathione S-transferase isoenzyme patterns in different subtypes of enzyme altered rat liver foci treated with the peroxisome proliferator nafenopin or with phenobarbital. AB - It is known that phenobarbital (PB) and the peroxisome proliferator (PP) nafenopin (NAF) promote tumor formation by stimulating selective growth of different subtypes of liver foci. While PB enhanced the gamma glutamyltranspeptidase (GGT)-positive eosinophilic-clear cell foci (ECF), NAF amplified the GGT-negative weakly basophilic foci (WBF). These findings provide the possibility of using the occurrence of these foci subtypes as early indicators for the carcinogenic potential of PB- and PP-type promoters. In order to improve the methods for the discrimination between ECF and WBF we studied further differences in their phenotype, as determined by the expression pattern of glutathione S-transferase (GST) subunits. GST subunits of the alpha (Ya, Yc), mu (Yb1, Yb2) and pi family (Yp), which compose different GST isoenzymes, were demonstrated by immunohistochemical methods. ECF were the only foci subpopulation that expressed GST subunit Yp, while this subunit was always absent in WBF and in another focus subtype, the tigroid foci (TF). Neither PB nor NAF changed this pattern. Thus Yp expression was rather a function of the focus type than of the promoter used. Upon PB treatment expression of the GST subunits Yb1 and Yb2 was frequently elevated in ECF, while Ya and Yc remained more or less unchanged. In NAF-treated livers large WBF, however, showed diminished expression of all investigated GST subunits of the alpha and mu family. In conclusion, PB seems to promote mostly ECF with elevated levels of mu and pi class GSTs, while low levels or absence of all GSTs tested may be associated with growth selection of WBF through the PP NAF. PMID- 8242873 TI - A reappraisal of the carcinogenicity of surface modified asbestos fibres. AB - A previous study using intrapleural administration of surface-modified amosite asbestos showed a difference in the number of pleural mesotheliomas induced with C18-hydrocarbon derivatised fibres compared to native amosite asbestos. The study has been repeated with larger groups of animals (30) under specific pathogen free conditions, resulting in an increase in the mean animal survival time for both fibre-treated groups. Under these conditions there was no significant difference between the numbers of pleural mesotheliomas induced by C18 hydrocarbon-modified amosite asbestos and native amosite asbestos. The major difference between the two studies was the mean time to death from tumour of rats exposed to fibres. The C18 amosite treated rats in the first study may not have had a mean survival time long enough to allow mesotheliomas to develop. PMID- 8242874 TI - Promotion of cholangiocarcinogenesis in the hamster liver by bile duct ligation after dimethylnitrosamine initiation. AB - Administration of hepatocarcinogenic nitrosamines before or after infection with the liver fluke, Opisthorchis viverrini (OV), results in marked development of cholangiocellular and hepatocellular precancerous and cancerous lesions in the hamster liver. The promoting effects of OV are believed to be exerted either mechanically, chemically or immunologically. To test the influence of possible mechanical effects, Syrian hamsters were initiated with a single i.p. injection of dimethylnitrosamine (DMN) 20 mg/kg and subjected 2 weeks later either to a sham operation or to complete ligation of the extrahepatic bile duct to the left lateral lobe. At the end of week 40, the animals receiving DMN-initiation and ligation had a 60.9% incidence of cholangiofibrosis, 21.7% of mucous cystadenomas and 39.1% of cholangiocarcinomas, whereas the group given DMN alone only developed cholangiofibrosis, limited to 5% of the animals. In the latter case neither cystadenomas nor cholangiocarcinomas were observed. The incidence of hepatocellular nodules did not differ between the two groups and no tumorous lesions developed in either the ligated or the untreated groups without DMN pretreatment. Complete ligation of the bile duct itself led to a series of events; obstruction of bile flow being followed by dilatation, cyst formation, and necrosis of the bile duct epithelium and surrounding affected areas leading to regenerative proliferation. The results are in line with the conclusion that parasite-associated proliferation in target cell populations is, at least in part, responsible for the influence of OV on liver tumor development. PMID- 8242875 TI - Evaluation of the post-initiation effects of oltipraz on aflatoxin B1-induced preneoplastic foci in a rat model of hepatic tumorigenesis. AB - Previous studies have demonstrated that ingestion of 5-(2-pyrazinyl)-4-methyl-1,2 dithiole-3-thione (oltipraz) during the aflatoxin B1 (AFB1) treatment phase completely prevented hepatic cancer. In this study we evaluated the effect of feeding oltipraz during the post-AFB1 treatment phase. Fifty-five male F344 rats were divided into five groups. All rats were gavaged with 25 micrograms AFB1/rat, five times a week for two successive weeks. The rats were fed the oltipraz supplemented diet according to three different feeding regimes: during the AFB1 treatment phase (1 week prior to, during and 1 week after the last gavage with AFB1); during the post-treatment phase; or throughout the entire time of the experiment. Phenobarbital-supplemented diet was fed during post-treatment phase to one group and this was used as a positive control for the promotion of AFB1 induced focal growth. The burden of putative, preneoplastic, hepatic glutathione S-transferase P-positive foci was evaluated at 13 weeks after the AFB1 treatment phase. As seen previously, oltipraz fed during the AFB1 treatment phase significantly inhibited focal development, i.e. the volume percent of the liver occupied with foci was reduced by 87%. Oltipraz when fed during the post treatment phase neither inhibited nor enhanced focal development. PMID- 8242876 TI - Partial hepatectomy of rats 3 weeks before or simultaneously with 2-aminofluorene injection can affect the amounts of adducts induced in hepatic DNA. AB - The influence of partial hepatectomy on the level of 2-aminofluorene (2-AF) induced DNA adducts in rat liver was studied. We found that partial hepatectomy performed either 3 weeks before or simultaneously with the injection of 2-AF affected the amounts of adducts in rat hepatic DNA compared to controls. The level of DNA adducts in rats that were treated with 2-AF and simultaneously hepatectomized was higher (19.9 fmol/microgram DNA) than in non-hepatectomized ones (14.4 fmol/microgram DNA) when measured 48 h after 2-AF administration. In rats treated with the carcinogen 3 weeks after hepatectomy the level of DNA adducts was significantly higher than in nonhepatectomized rats when measured 15 days after the injection of 2-AF (10.9 fmol/microgram DNA and 5.9 fmol/microgram DNA respectively). The high level of DNA adducts in that group of hepatectomized animals was correlated with a relatively lower rate of 2-AF-induced radioactive thymidine incorporation into hepatic DNA (in comparison to non-hepatectomized rats). PMID- 8242877 TI - Effect of ascorbate and 5-aminosalicylic acid on light-induced 8 hydroxydeoxyguanosine formation in V79 Chinese hamster cells. AB - Recently we showed that ascorbate and 5-aminosalicylic acid (5-ASA) prevented 8 hydroxydeoxyguanosine (8-OHdG) formation in calf thymus DNA exposed to UV-visible light. However, the ultimate defense against oxidative DNA damage depends on an intracellular/intranuclear effect of the compounds. In the present study we investigated the effect of ascorbate and 5-ASA on 8-OHdG formation in V79 Chinese hamster cells exposed to light from a sun-lamp. Exposure for 1 min (4560 mJ/cm2) increased 8-OHdG formation in cellular DNA to 30-40 times background level. Preincubation of the cells with ascorbate or 5-ASA at concentrations of 0.1, 1 and 10 mM diminished the 8-OHdG formation to 0.67, 0.74 and 0.49 times controls (P < 0.05) for ascorbate respectively, and to 0.82, 0.66 and 0.33 times controls (P < 0.05), for 5-ASA. These findings demonstrate that both ascorbate and 5-ASA prevent oxidative DNA damage in cells by acting as intracellular/intranuclear antioxidants. PMID- 8242878 TI - Peroxisomal enzyme induction uncoupled from enhanced DNA synthesis in putative preneoplastic liver foci of rats treated with a single dose of the peroxisome proliferator nafenopin. AB - Putative preneoplastic foci of spontaneous origin could be detected in the livers of 2 year old, untreated male Wistar rats. The unaltered and preneoplastic hepatocytes showed an identical expression of the peroxisomal marker enzyme acyl CoA oxidase, as determined by immunohistochemical staining. A single dose of the peroxisome proliferator (PP) nafenopin (NAF) induced the enzyme predominantly in hepatocytes around the central venules and cell replication mainly in the periportal areas. However, upon one NAF application almost all of the preneoplastic foci showed a considerably weaker immunoreaction for peroxisomal acyl-CoA oxidase than the surrounding tissue. Concomitantly NAF elevated replicative DNA synthesis index in foci up to approximately 40%, while replication of hepatocytes in the unaltered portion of the livers increased only slightly to moderately. In conclusion, NAF-induced peroxisomal acyl-CoA oxidase and replicative DNA synthesis seem not to be necessarily coupled within the same liver cell. Furthermore, preneoplastic foci responded rather to the cell replicative than to the peroxisomal effects of NAF, suggesting that the PP induced growth stimulus is of particular significance for the carcinogenic action of this class of compounds. PMID- 8242879 TI - Effects of hypertonic saline solution (20%) on cardiodynamics during hemorrhagic shock. AB - The cardiodynamic effects of using hypertonic saline solution were analyzed in dogs after inducing hemorrhagic shock. Administration of 1.5 mL/kg 20% sodium chloride resulted in circulatory improvement: 200% increase in blood pressure, 260% increase in cardiac output, 60-70% increase in pulmonary arterial pressure, 66% increase in pulmonary wedge pressure, 200% increase in central venous pressure, 33% decrease in total peripheral resistance, and 40% decrease in pulmonary vascular resistance. The effects of sodium chloride on cardiodynamics probably resulted in elevation of preload and reduction of afterload. PMID- 8242880 TI - Coronary vascular function after hemorrhagic hypotension in dogs. AB - This study tested the hypothesis that hemorrhagic hypotension alters intrinsic contraction-relaxation mechanisms of coronary arteries. Coronary vascular smooth muscle (VSM) was evaluated ex vivo using left circumflex coronary artery preparations isolated from beagle dogs 4 hr after sham hemorrhage (controls) or maintained hemorrhagic hypovolemia. Hemorrhaged dogs exhibited systemic hypotension (mean arterial pressure approximately 65 mm Hg), tachycardia, and tachypnea during the 4 hr in vivo phase of the study, accompanied by 30-50% reductions in left ventricular myocardial blood flows (P < 0.05). Coronary arteries isolated from these dogs were stretched to the asymptote of their length contractile tension relationship; no significant differences were observed in length-active tension or length-passive tension relations between hemorrhage and control arteries. Similarly, neither the maximal responses nor the EC50 values for isometric contractions produced by prostaglandin F2 alpha (PGF2 alpha) (10( 8) to 3 x 10(-5) M) or depolarizing concentrations of K+ (10-100 mM) were altered by hemorrhage (P > 0.05). Vasodilator responses to the cyclic guanosine monophosphate (GMP)-dependent VSM relaxant nitroprusside (10(-4) M) also were not prevented by the hemorrhage protocol. In contrast, coronary VSM relaxation induced by the endothelium-dependent vasodilator acetylcholine (10(-9)-10(-5) M) was significantly decreased by 25-50% in K(+)- and PGF2 alpha-precontracted coronary arteries from the hemorrhaged dogs (P < 0.01). We conclude that receptor (PGF2 alpha)-dependent and membrane depolarization (K+)-dependent contractile mechanisms remained operational in coronary arteries during hemorrhagic hypotension, as did basal cyclic GMP-dependent VSM relaxation mechanisms. However, diminution of acetylcholine-induced relaxation of coronary VSM suggests impaired endothelium-dependent vasodilation in the coronary vasculature during acute (4 hr) hemorrhagic hypotension. PMID- 8242881 TI - Alterations in renal gluconeogenesis and blood flow during hemorrhagic shock. AB - The role of renal gluconeogenesis following hemorrhagic shock was studied. Hemorrhagic shock was induced in fasted, anesthesized rats by reduction of blood pressure to 40 mm Hg for either 30 or 150 min. Plasma samples were obtained for plasma glucose determination, and right renal arterial blood flow was determined with the help of an ultrasonic transit time blood flowmeter in hemorrhagic shock and control rats. The kidney was perfused via the renal artery with 37 degrees C oxygenated, glucose-free Krebs-Henseleit solution in the presence of 100 microM phloridzin to inhibit the cellular reuptake of glucose. Renal glucose production was determined by measuring glucose in both renal vein effluent and urine. After 30 min of equilibration, 5 mM lactate and 0.5 mM pyruvate were added to the perfusate as a gluconeogenic substrate, and effluent samples were collected after 5, 10, and 15 min. Moderate hyperglycemia was observed in vivo following 30 min of hemorrhagic shock, and significant hypoglycemia (P < .05) was observed following 150 min of hemorrhagic shock. Renal arterial flow was significantly decreased at 30 min (P < .05) and 150 min (P < .05) of hemorrhagic shock. Renal glucose production with and without substrate after 30 min of hemorrhagic shock was similar compared to control. Renal glucose production after 150 min of hemorrhagic shock was significantly decreased (P < .05) compared to control.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242882 TI - Differences in arterial and arteriolar endothelial structure during endotoxin shock. AB - Previous studies of physiological and ultrastructural assessment changes in the walls of the femoral artery and A1, A2, and A3 arterioles in the rat cremaster muscle after infusion of Escherichia coli endotoxin (ENDT) (6 mg/kg-1 hr. period) indicate there may be a difference in the alteration of the endothelial structure of arteries and arterioles. Functionally, ENDT has been shown to abolish acetylcholine (ACh)-induced relaxation in the femoral artery [Zhou, PhD dissertation, University of South Florida, Tampa, 1992]. Dilations of A1 arterioles were shown to be reduced, but dilations of A2 and A3 arterioles were not significantly changed from control [Baker and Sutton, Am J Physiol, 264:H 1118-H 1123, 1993]. In the current study, ultrastructural evaluation of femoral arterial tissue post-ENDT at the mean arterial pressure (MAP) of 100 mm Hg revealed partially destroyed endothelial cells. The MAP decreased as the animal progressed into shock. At 80 mm Hg, 60 mm Hg and 40 mm Hg, essentially all endothelial cells were destroyed, with the internal elastic lamina denuded. In contrast, endotoxin damage in the A1, A2, and A3 arterioles was minimal even at a MAP of 40 mm Hg. Endothelial cells of A1 arterioles post-ENDT had more vacuoles than at control. Therefore, arteriolar endothelium was functionally and anatomically relatively undamaged, consequent to endotoxin administration, in contrast to femoral arteries where the endothelium was destroyed in a short time. PMID- 8242883 TI - Evidence of increased nitric oxide production in patients with the sepsis syndrome. AB - To investigate whether the hypotension of septic shock is due to an excess production of nitric oxide (NO), we have determined the serum levels of nitrate and nitrite (NO3/NO2), the stable end-products of NO, in 12 patients with the sepsis syndrome and marked hypotension. Compared to a mean NO3/NO2 level of 36.4 microM in controls (n = 7), the group of septic patients had a significantly elevated mean NO3/NO2 level of 124 microM (P < 0.01, Wilcoxon two-sample test). A lesser elevation was also seen in a group of postoperative patients (mean level 87.3 microM, n = 7), which was significantly elevated compared to controls (P < 0.01, Wilcoxon two-sample test), but was not significantly lower than the septic group (0.1 > P > 0.05, Wilcoxon two-sample test). These data suggest that NO may be responsible, at least in part, for the hypotension of septic shock. PMID- 8242884 TI - Effect of subcutaneous Escherichia coli-induced hypermetabolic sepsis on hepatic gluconeogenesis and its hormonal responsiveness in the rat. AB - In hypermetabolic sepsis, gluconeogenesis is markedly elevated during fasting, and is manifested as an increased rate of glucose appearance (Ra). The likely causes of such a change are alterations in 1) concentration of systemic hormones, 2) concentration of glucose precursors, especially lactate, 3) activity of the key enzymes of the pathway, and 4) hormone receptors and/or transmembrane signalling mechanisms, involved in the hormonal regulation of the pathway. In this study, we investigated the importance of the latter two factors in the increase of gluconeogenesis during hypermetabolic sepsis. Rats were rendered septic by repeated subcutaneous administration of live Escherichia coli. The livers were perfused in vitro in a nonrecirculating mode to measure the rate of gluconeogenesis from saturating concentrations of lactate (5 mM) or lactate (5 mM) + pyruvate (0.5 mM), and the response of gluconeogenesis to vasopressin (VP, 0.1 and 1.0 nM), glucagon (Glc, 0.1 and 1.0 nM), and prostaglandin (PG) F2 alpha (5 microM). The rate of gluconeogenesis without precursor supply was approximately 20-30 mumoles/100 g b w/hr during the first 4-6 min of perfusion, followed by a continuous decline to very low levels. Infusion of lactate (5 mM) or lactate (5 mM) + pyruvate (0.5 mM) increased glucose output, and maintained it at approximately 100-110 and approximately 130-140 mumoles/100 g b w/hr, respectively. VP, Glc, and PGF2 alpha stimulated the rate of gluconeogenesis in a dose-dependent manner (VP and Glc). No differences were observed between control and septic rats using these stimuli.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242885 TI - Retrospective study comparing the pathophysiology of antibiotic-treated and untreated Escherichia coli- and Staphylococcus aureus-infused baboons. AB - Escherichia coli and Staphylococcus aureus are the most common pathogens encountered in septic shock. This is a descriptive study in which the pathophysiologic response to infusions of LD100 concentrations of E. coli and S. aureus are staged and compared. Equivalent concentrations of both organisms were infused over a 2 hr period into antibiotic-treated and untreated animals with the following results: 1) The apparent clearance of E. coli was less than that of S. aureus over the 2-hr infusion period, but far greater during the next 8 hr in both antibiotic-treated and untreated animals. Thus the clearance of E. coli fits a one-compartment (intravascular), and that of S. aureus fits a two-compartment (intra- and extravascular) model. 2) The intensity of the cardiovascular, temperature, and metabolic response to E. coli was greater, whereas that of the disseminated intravascular coagulant (DIC) response to S. aureus was greater. We conclude, therefore, that the response to E. coli consists of four stages with no invasion and colonization of tissues, whereas the response to S. aureus consists of two stages with invasion and colonization of tissues. PMID- 8242886 TI - Troponin-T and CK MB (mass) in early diagnosis of ischemic myocardial injury. The Helsingborg Study, 1992. AB - We evaluated cardiac troponin T (S-troponin T) versus CK MB in serum (S-CK MB) sampled 4, 10, 16, and 22 h after onset of acute symptoms in 207 consecutive cases admitted to our coronary care unit in Helsingborg, Sweden, May-October 1992. These were primarily classified into 106 acute myocardial infarctions (AMI) and 101 NOT-AMI cases by conventional cardiological criteria plus S-CK MB (mass). Time curves of S-troponin T and S-CK MB data were plotted for each individual case. Twelve of the 101 cases in the NOT-AMI group showed increased S-troponin T indicating ischemic myocardial injuries. The same cases also showed changes of S CK MB (mass), though below its discriminator. Seven of these cases were reclassified as minor myocardial damage, constituting 25% of our 28 cases of unstable angina. The remaining five cases showed a combination of constantly increased S-troponin T and decreasing or low S-CK MB mass values as seen after a recent infarction. Consequently, the patient material was reclassified into 118 cases of ischemic myocardial injury (106 conventional AMI + 7 minor myocardial damage + 5 postinfarctions) and 89 cases of NOT-ischemic myocardial injury. The frequency distributions of the maximal S-troponin T and S-CK MB (mass) values of each case were plotted in double test evaluation histograms. For troponin T, discriminator 0.20 micrograms/L, clinical sensitivity for ischemic myocardial injury was 97% and specificity 99%. With a lower discriminator of 0.10 micrograms/L, sensitivity increased to 99% and specificity decreased to 89%. For S-CK MB (mass), discriminator 10 micrograms/L, sensitivity was 91%, specificity 98%. With a lower discriminator of 5 micrograms/L sensitivity increased to 96% and specificity decreased to 78%. We conclude that as a single routine test, S troponin T is the marker of choice for early diagnosis of ischemic myocardial damage. The combination S-troponin T and S-CK MB (mass) provides additional, detailed information in reinfarction and postinfarction unstable angina pectoris. PMID- 8242887 TI - Enzyme diagnosis of liver diseases. PMID- 8242888 TI - Biochemical tests in the diagnosis of chronic pancreatitis and in the evaluation of pancreatic insufficiency. AB - Chronic pancreatitis (adults) and cystic fibrosis (children) are the most common diseases leading to exocrine pancreatic insufficiency that, when reduced to < 5% of normal function, is characterised by steatorrhoea. The pathogenesis of the former condition is outlined, and recent concepts are emphasized. Biochemical tests to detect pancreatic insufficiency and to identify pancreatic disease as the cause of steatorrhoea include: serum enzyme tests (lipase, amylase, trypsin); stool chymotrypsin; isotopic tests based upon the assimilation of [14C] lipids and starch or excretion of the isotope as breath CO2, as well as the dual labelled Schilling test; oral function tests utilising substrates hydrolysed by pancreatic enzymes such as benzoyl tyrosyl-p-aminobenzoic acid and fluorescein dilaurate; and duodenal intubation studies following meal-induced or hormonal stimulation of the pancreas. The rationale for these tests and the cumulative clinical experience of their utility are reviewed. A recommended diagnostic strategy is briefly presented. The role of various biochemical procedures to evaluate the efficacy of pancreatic enzyme replacement therapy is also described. PMID- 8242889 TI - Clinical significance of microalbuminuria and evaluation of the Micral-Test. AB - Microalbuminuria is defined as excretion of albumin between 20 and 200 micrograms/min. Persistent microalbuminuria indicates a high probability of damage to the glomerular filtration capacity of the kidney and is of great diagnostic relevance: (a) in diabetes, for early diagnosis of diabetic nephropathy; (b) in patients with hypertension, as an indicator of end-organ damage associated with a lowered life expectancy; (c) in pregnancy, as a possible predictor of developing preeclampsia. For screening, a concentration of 20-200 mg/L of albumin in the first morning urine has been proven to be a suitable indicator. The Micral-Test is a test strip now available that makes a semiquantitative assessment of the albumin concentration in the urine at various levels (0, 10, 20, 50, 100 mg/L). Many studies have shown that the sensitivity and specificity of the test strip is 90% or higher in urine containing elevated albumin concentrations. Furthermore, it was shown that there is no influence on the measurement from interfering factors such as glucose concentration, pH value, ketonuria, storage of the sample, or bacterial contamination in the urine. The Micral-Test offers a simple, exact, and with respect to interference effects, a robust testing method for the semiquantitative assessment of albumin concentration in the urine. PMID- 8242890 TI - Toward routine diagnosis of hepatitis B virus desoxyribonucleic acid. AB - Although a multitude of hepatitis B markers are available to discriminate different states of hepatitis B virus (HBV) infection, there remains a need to detect the HBV DNA itself. The HBV DNA test is the only test that can quantify the number of complete virions. As shown in this review, the diagnostic value of testing for HBV DNA depends on quantification, and there are situations in which in vitro amplification is necessary and others in which testing for HBV DNA without preamplification is best. Indications for analysis of HBV DNA include the early recognition of chronic hepatitis B, discrimination of different HBsAg carrier states, recognition of the evolution of variants, indication and follow up of anti-viral therapy with interferon, and assessment of blood products for infectivity. In each of these cases quantification of HBV DNA improves the diagnosis of HBV infection and will become an important tool in the routine diagnosis of hepatitis B virus infection in the future. PMID- 8242891 TI - The possible link between insulin dependent (juvenile) diabetes mellitus and dietary cow milk. PMID- 8242892 TI - The mechanism of insulin action. AB - Insulin was isolated over 70 years ago, but the intracellular transduction of the insulin signal has not been elucidated. Significant progress has been made, particularly in the last 10 years, with the characterization of the insulin receptor and its intrinsic tyrosine kinase. However, no mechanism has been proposed that accounts for all the actions of insulin. Furthermore, all the mechanisms discussed in this brief overview contain major inadequacies. Despite these gaps in our knowledge, substantial evidence indicates that receptor tyrosine kinase activity is essential for insulin action and multiple pathways almost certainly participate. Hopefully, continued dissection of the signalling pathways will soon yield the mechanism of insulin action. PMID- 8242893 TI - Detection of progressive diabetic nephropathy: role of microalbuminuria determination. PMID- 8242894 TI - Good and bad diabetes. PMID- 8242895 TI - Prevention and cure of juvenile diabetes. In our lifetime? AB - Advances in our understanding of the pathophysiology of IDDM have allowed us to consider potentially curative therapies for IDDM. A number of interventional strategies may be effective in halting beta-cell destruction and preventing the clinical expression of IDDM. Successful implementation of these strategies will be dependent upon the identification of individuals at high risk for developing IDDM. Similarly, technological advances in immunoisolation technology and porcine islet isolation may allow transplants for large numbers of diabetics sufficiently early in the disease to prevent complications. All of these developments make it likely that prevention and cure of IDDM will be possible in our lifetime. PMID- 8242896 TI - Diabetic nephropathy: are we making progress in delaying renal failure? PMID- 8242897 TI - Biochemical diagnosis and monitoring of diabetes: a laboratory perspective for the nineties. PMID- 8242898 TI - Low NK syndrome and its relationship to chronic fatigue syndrome. PMID- 8242899 TI - Roles of soluble fibronectin and beta 1 integrin receptors in the binding of Mycobacterium leprae to nasal epithelial cells. AB - The mechanisms by which Mycobacterium leprae invades the human host are presently unknown. We investigated the ability of M. leprae to bind to human RPMI 2650 cells, a human nasal septal epithelial cell line, using both microscopic observation and an ELISA technique. The results demonstrated that M. leprae adheres to nasal cells after binding to soluble fibronectin. Furthermore, it was observed that M. leprae could bind to the beta 1 chain of the integrins in the absence of serum or mucus. These results demonstrated that M. leprae uses fibronectin and fibronectin receptors on the surface of epithelial cells to bind and possibly invade the nasal epithelial cells. PMID- 8242900 TI - Murine experimental autoimmune orchitis (EAO) induced by syngeneic testicular germ cells (TC) alone: orchitogenic and lymphostimulatory activities of TC from mice at various ages. AB - As part of our continuous investigations on the immune pathogenesis of a new experimental autoimmune orchitis (EAO) model without the use of adjuvant in C3H/He mice, the developmental patterns of autoantigenicity of viable syngeneic testicular germ cells (TC) during the postnatal period were investigated by examining the orchitogenic and immunogenic activities of TC, the lymphostimulatory activities of TC (the cytokine release from sensitized spleen cells (SPC) in response to TC), and the immunohistochemical localization of autoantigens in the testes of normal mice at various weeks of age. TC from 6-week old mice had the capability of inducing EAO (orchitogenicity) for the first time. Delayed-type hypersensitivity-inducing capacity and anti-TC antibody-eliciting capacity were initially observed in mice immunized with TC from 4-week-old mice. A significant stimulation of interleukin 5 (IL-5) and interleukin 6 (IL-6) production by sensitized SPC was detectable when the TC from 3-week or older mice were employed as a stimulant. Interleukin 2 (IL-2) and interferon-gamma (IFN gamma) production was detected with TC from 4-week or older mice. Immunohistochemical staining reaction with anti-TC antisera was primarily localized at the acrosomal portion of spermatids and spermatozoa in the seminiferous tubules and was detected in early and late spermatids of mice as early as 3 weeks. Thus, from these chronological data it is suggested that the appearance of the lymphostimulatory activities of TC consistently precedes that of the orchitogenic activity and that the production of IL-5 and IL-6 (Th 2 cytokine) by sensitized SPC upon in vitro TC stimulation is detectable earlier than the production of IL-2 and IFN-gamma (Th 1 cytokine) in the postnatal period. PMID- 8242901 TI - Systemic and mucosal IgA responses to systemic antigen challenge in IgA nephropathy. AB - IgA nephropathy (IgAN) is characterized by the deposition of glomerular IgA. The source of the deposited IgA is not known, but both the mucosal and systemic IgA systems have been implicated. In order to investigate mucosal and systemic antibody production to systemic antigen challenge in IgAN, 20 patients and 20 controls where immunized with tetanus toxoid (TT). While patients with IgAN responded with a similar serum IgG, IgA, IgA1, and IgA2 antibody response to controls, they did, however, produce more IgA1 antibodies relative to IgA2 (P < 0.05). No salivary IgA antibody response was observed to systemic immunization in controls; however, there was a significant IgA response to TT in the saliva of patients with IgAN. IgA antibodies were produced in vitro by Epstein Barr virus (EBV)-transformed peripheral blood lymphocytes (PBLs) obtained from control blood only when taken shortly (1 or 2 weeks) after immunization. Patients with IgAN produced significantly more IgA anti-TT positive cultures than controls and for a longer period (P < 0.01) after immunization. In contrast, IgG anti-TT was produced in EBV-transformed cultures at all time points, but with no difference between IgAN and controls in the proportion of IgG producing cultures. These results demonstrate increased IgA antibody production in both the systemic and mucosal IgA systems following systemic immunization in IgAN and suggest an abnormal overlap between the two systems in IgAN. PMID- 8242902 TI - The effect of rapamycin on c-jun expression in human lymphocytes. AB - Rapamycin (RAPA) is a potent immunosuppressant. Several reports indicate that the drug can act at the late G1 stage of the lymphocyte activation. We studied the effect of RAPA on the expression of an immediate early phase gene c-jun, which plays a pivotal role in cell activation. The results showed that RAPA could inhibit PHA-induced c-jun expression by human T cells. This strongly suggests that there exists a mechanism for RAPA to interact with the lymphocyte activation cascade at a very early stage. We also demonstrated that in T cells a Na+/K+ ATPase inhibitor, ouabain, could induce a late (16 h poststimulation) c-jun expression, which was sensitive to cyclosporin A (CsA) but not to RAPA. This suggests that c-jun's role is probably not restricted to the early phase of lymphocyte activation. PMID- 8242903 TI - Prevention of type I diabetes with lymphotoxin in BB rats. AB - We previously reported that nonspecific immunomodulations with a streptococcal preparation (OK-432), an inducer of tumor necrosis factor (TNF), or with recombinant TNF prevented development of insulin-dependent diabetes mellitus (IDDM) in animal models (NOD mice and BB rats). Recently we have further reported that lymphotoxin (LT), a cytokine with functional and structural characteristics similar to those of TNF, also protected NOD mice from diabetes. In this study, we have extended our observation on the LT to BB rats. Male and female BB/Wor rats were treated intraperitoneally with recombinant human LT three times a week from 4 to 11 weeks of age. The cumulative incidence of diabetes by 14 weeks of age was 24/30 (80.0%) in nontreated control rats, whereas it was 10/26 (38.5% vs control, P < 0.01) and 4/29 (13.8% vs control, P < 0.0001) in the rats treated with 1 x 10(3) and 1 x 10(4) of LT, respectively. There was no significant difference in nonfasting blood glucose levels and body weights between nontreated control and LT-treated rats, which were nondiabetic. In the LT-treated rats, intensity of insulitis was significantly reduced in comparison with the nontreated rats. Concanavalin A-stimulated TNF/LT productivity of spleen cells was significantly lower in BB/Wor and BB/Sendai rats than in Wistar rats or other normal rat strains. On the other hand, there was no difference between BB/Sendai and Wistar rats in the in vivo TNF/LT productivity induced with LPS or with IFN-gamma plus LPS, and the TNF/LT productivity of these rats was lower on stimulation with LPS alone, but higher with IFN-gamma plus LPS than the other normal rats. These results indicate that treatment with LT, as well as TNF, modulated autoimmunity and prevented development of IDDM in BB/Wor rats which may be low producers of TNF/LT. PMID- 8242904 TI - Murine complement-mediated immune clearance dysfunction is associated with the lymphoproliferative (lpr) gene. AB - Using a branched series model of immune clearance and an iterative curve fitting process, we have previously demonstrated abnormal in vivo clearance of opsonized erythrocytes in autoimmune MRL-lpr/lpr mice. This technique allows simultaneous evaluation of four rate constants governing both complement- and Fc-mediated clearance; i.e., complement-mediated sequestration (k1), C3b deactivation and release (k2), complement-dependent phagocytosis (k4), and Fc gamma-mediated sequestration and phagocytosis (k3). To evaluate genetic factors which may contribute to abnormal clearance in MRL-lpr/lpr mice, serial clearance studies were performed in congenic MRL-(+/+) mice, as well as in several mouse strains homozygous for the lymphoproliferative gene; i.e., BALB/c-lpr/lpr and C57BL/6 lpr/lpr mice. Rate data were obtained from mice 3 months through 12-18 months of age, and mean rate constant values compared to control BALB/c and/or C57BL/6 mice. Clearance of opsonized cells in MRL-lpr/lpr mice was characterized by both abnormal complement and Fc gamma receptor function with decreased complement mediated sequestration, complement-dependent phagocytosis, and Fc gamma-mediated sequestration and phagocytosis. In contrast, abnormal clearance in congenic MRL (+/+) mice was characterized by decreased Fc gamma-mediated sequestration and phagocytosis (P < 0.0001) only. Both complement-mediated sequestration and complement-dependent phagocytosis were significantly decreased in BALB/c-lpr/lpr and C57BL/6-lpr/lpr mice (P < 0.0001), with the pattern and magnitude of abnormal complement clearance kinetics similar to that occurring in MRL-lpr/lpr mice and contrasting with normal complement-mediated clearance in control BALB/c and C57BL/6 mice. Since decreased complement-mediated clearance was observed in three differing strains of mice homozygous for the lymphoproliferative gene, these data suggest that clearance of opsonized cells in mice is in part genetically determined, and that there may be a specific association between complement mediated clearance dysfunction and the murine lymphoproliferative gene. PMID- 8242905 TI - Monoclonal anti-idiotypic antibodies against human anti-thyroglobulin autoantibodies recognize idiotopes shared by disease-associated and natural anti thyroglobulin autoantibodies. AB - Monoclonal murine anti-idiotypic antibodies generated against purified human anti thyroglobulin (TG) antibodies from a patient with Hashimoto's thyroiditis recognized two major clusters of idiotopes (1C2 and 1B1) on human anti thyroglobulin antibodies which were framework and paratope-related. A survey of affinity-purified autoantibodies from Hashimoto's patients indicated that the 1C2 and 1B1 idiotopes are expressed on the disease-associated anti-TG antibodies from several patients. 1C2 and 1B1 idiotopes are also detected on affinity-purified anti-TG antibodies isolated from serum of healthy donors and with therapeutic preparations of immunoglobulins obtained from plasma pools of healthy individuals. The observation that both framework and paratope-related idiotopes are shared by natural and patients' autoantibodies suggests that the anti-TG autoantibodies found in sera of patients with Hashimoto's disease represent an abnormal expansion of normal autoreactive B cell clones rather than the emergency of an antigen-driven, disease-specific B cell response. PMID- 8242906 TI - Myocardial infarction, thrombolytic therapy, and economics. PMID- 8242907 TI - Thrombolytic therapy for unstable angina. AB - Thrombolytic therapy for unstable angina has not gained acceptance as a primary treatment for unstable angina (UA) despite the evidence showing a reduction in mortality when these agents are given for myocardial infarction. The purpose of this review is to examine the clinical value of thrombolytic therapy for UA. The multiple lines of evidence supporting intracoronary thrombus formation as a key mechanism in the pathogenesis of UA are reviewed. Studies examining the effect of thrombolytic therapy on angiographic endpoints have shown little effect on the extent of luminal narrowing, but do reveal a decrease in angiographically detected thrombus. Twelve randomized, controlled trials of thrombolytic agents in 611 UA patients with predefined clinical endpoints have been published. These trials varied widely in design and adjunctive therapy both in treated and control grops. Review of these trials show a tendency to fewer clinical events such as death, infarction, and need for revascularization in treated patients, with a corresponding increase in bleeding complications. Clinical efficacy of thrombolytic therapy cannot be excluded by the available data, perhaps in part because of insufficient numbers of patients treated. Determination of the net clinical value of thrombolytic therapy must await larger and more definitive trials. PMID- 8242908 TI - Patterns of radiofrequency catheter ablation of left free-wall accessory pathways: implications for accessory pathway anatomy. AB - Despite the abundance of literature on the electrophysiology of accessory pathways, clinical data on their anatomic properties remain infrequent. The small and discrete nature of lesions generated by radiofrequency (RF) energy may allow better characterization of accessory pathway anatomy in the intact heart. RF catheter ablation was performed on 40 left free-wall accessory pathways in 39 consecutive patients with a unipolar endocardial approach. The patterns of accessory pathway ablation were identified. Spatial-electrophysiologic information provided by the ablation catheter at individual sites of RF application and corresponding data from the coronary sinus catheter were correlated with the effects of RF energy on accessory pathway conduction. Of 39 accessory pathways permanently (n = 37) or transiently (n = 2) ablated, 24 had "simple" ablation, with abolition of conduction by one individual RF application. In 15 of 24 pathways that could be crossed by the coronary sinus catheter, the concordance in anatomic and electrophysiologic information between the site of earliest retrograde atrial activation and the effective ablation position (ventricular approach) suggested a perpendicular fiber course. Fifteen pathways had "complex" ablations; of these, eight had spatial-electrophysiologic discordance between the atrial and ventricular insertions, suggesting an oblique fiber orientation. Seven pathways had modification or transient suppression of conduction, with or without subsequent abolition of conduction at identical or physically disparate (> 1 cm apart) sites; four pathways had sequential ablation of antegrade and retrograde conduction. These raised possibilities of broad fiber span and functional longitudinal dissociation of accessory pathway conduction. Accessory pathways with simple and complex ablations did not differ in clinical and electrophysiologic parameters. Complex ablations demanded more lengthy and difficult procedures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242909 TI - Silent ischemia and loss of reversible myocardial dysfunction following myocardial infarction. AB - Sixty-seven asymptomatic patients were enrolled after a first uncomplicated myocardial infarction (MI) so as to study the relevance of reversible myocardial dysfunction in determining left ventricular function soon after the acute episodes and 12 months later. Moreover, the potential role of silent ischemia in conditioning the evolutive aspects of contractile dysfunction has been investigated. Postextrasystolic potentiation during two-dimensional echocardiographic (2-D echo) monitoring has been used to detect the presence of viable myocardium in asynergic myocardial segments. Results of electrocardiographic (ECG) ambulatory monitoring at predischarge determined patient groups: Group A included 49 patients without ST changes during monitoring, while Group B included 18 patients with silent ischemia. Incidence of reversible myocardial dysfunction was similar in the two study groups (82 vs. 86%, p = NS). Group B patients were older (59.6 +/- 6.7 vs. 50.6 +/- 10.6 years, p < 0.015) and had lower ejection fractions (EFs, 43.4 +/- 6.4% vs. 51.2 +/- 8.3%, p = 0.026) and higher at-rest wall-motion scores (WMSs, 11.4 +/- 5.9 vs. 7.2 +/- 3.8, p = 0.019). Left ventricular end-diastolic volume (LVEDV) and potentiated WMS did not differ. At 1-year examination, Group B patients exhibited a greater LVEDV index (96 +/- 6.5 vs. 70.7 +/- 14 ml/m2, p < 0.002) with a worsening both in rest and in potentiated wall-motion score index (12.8 +/- 4.6 vs. 5.3 +/- 1.8, p < 0.001; 9.2 +/- 3.6 vs. 4.8 +/- 2.2, p < 0.001, respectively). Left ventricular EF remained significantly depressed in Group B patients (42 +/- 8.7% vs. 55.5 +/- 8.1%, p < 0.002).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242910 TI - Cutting balloon coronary angioplasty--initial clinical experience. AB - The success of percutaneous transluminal coronary angioplasty is limited by acute occlusion and late restenosis. In 25 patients (20 men, 5 women, age range 36-81 years) coronary angioplasty was performed using a new cutting balloon into which 3-4 longitudinally orientated blades are incorporated so as to reduce the rate of severe dissections. In 12 patients stenoses were reduced from 83.9 +/- 7.8% to 28.4 +/- 10.7% (mean +/- SD) by the cutting balloon alone, using predilatation with a small conventional balloon in two cases. Thirteen other patients were additionally dilated with a conventional balloon because of a residual stenosis > 50% after cutting balloon angioplasty. Here the stenoses could be reduced from 78.1 +/- 8.7% to 29.1 +/- 11.3%. Six months follow-up angiography in 14 patients showed > 50% restenosis in two of seven patients dilated with a conventional balloon in addition to the cutting balloon, and in one of seven patients dilated with the cutting balloon alone but predilated with a small conventional balloon. These results show that coronary angioplasty by the new cutting balloon results in a stenosis reduction comparable with conventional balloons at a low complication rate. Available 6 months follow-up data show three restenoses in patients either pre- or postdilated by a conventional balloon and none in stand alone cutting balloon cases. PMID- 8242911 TI - Diagnosis of papillary muscle rupture after acute myocardial infarction by transthoracic and transesophageal echocardiography. AB - The sensitivity of transthoracic echocardiography to visualize the structural abnormality of papillary muscle rupture (PMR) after acute myocardial infarction can be anticipated to average about 50%; therefore, we evaluated five patients exhibiting the condition with both transthoracic and transesophageal echocardiography. The use of the two imaging techniques resulted in the fact that no instance of PMR was missed. Using transthoracic echocardiography in two patients and transesophageal echocardiography in four, the ruptured papillary muscle was visualized directly. Mitral insufficiency as an indirect sign was observed in all patients. In one patient the papillary muscle rupture developed in a mitral valve previously affected by endocarditis. All patients underwent mitral valve replacement and coronary artery bypass grafting. The diagnosis was confirmed at surgery in all patients. Four patients died in hospital, the fifth 5 months later. We recommended that transesophageal echocardiography be performed in patients with suspected PMR if transthoracic echocardiography does not provide an unequivocal diagnosis. PMID- 8242912 TI - Electrocardiogram signal variance analysis in the diagnosis of coronary artery disease--a comparison with exercise stress test in an angiographically documented high prevalence population. AB - Variance electrocardiography (variance ECG) is a new resting procedure for detection of coronary artery disease (CAD). The method measures variability in the electrical expression of the depolarization phase induced by this disease. The time-domain analysis is performed on 220 cardiac cycles using high-fidelity ECG signals from 24 leads, and the phase-locked temporal electrical heterogeneity is expressed as a nondimensional CAD index (CAD-I) with the values of 0-150. This study compares the diagnostic efficiency of variance ECG and exercise stress test in a high prevalence population. A total of 199 symptomatic patients evaluated with coronary angiography was subjected to variance ECG and exercise test on a bicycle ergometer as a continuous ramp. The discriminant accuracy of the two methods was assessed employing the receiver operating characteristic curves constructed by successive consideration of several CAD-I cutpoint values and various threshold criteria based on ST-segment depression exclusively or in combination with exertional chest pain. Of these patients, 175 with CAD (> or = 50% luminal stenosis in 1 + major epicardial arteries) presented a mean CAD-I of 88 +/- 22, compared with 70 +/- 21 in 24 nonaffected patients (p < 0.01). Variance ECG provided a stochastically significant discrimination (p < 0.01) which was matched by exercise test only when chest pain variable was added to ST segment depression as a discriminating criterion. Even then, the exercise test diagnosed single-vessel disease with a significantly lower sensitivity. At a cutpoint of CAD-I > or = 70, compared with ST-segment depression > or = 1 mm combined with exertional chest pain, the overall sensitivity of variance ECG was significantly higher (p < 0.01) than that of exercise test (79 vs. 48%). When combined, the two methods identified 93% of coronary angiography positive cases. Variance ECG is an efficient diagnostic method which compares favorably with exercise test for detection of CAD in high prevalence population. PMID- 8242913 TI - Congestive heart failure in a patient with systemic hypertension. AB - A 70-year-old man developed hypertension many years previously and had a bout with severe congestive heart failure 4 to 5 years before his death. Autopsy showed congestion and edema characteristic of heart failure, and enlarged heart, and slight focal interstitial left ventricular fibrosis, but only slight to moderate coronary atherosclerosis. PMID- 8242914 TI - Coronary artery bypass grafting for left main trunk coronary artery lesion associated with essential thrombocythemia. AB - Effort angina due to left main trunk (LMT) lesion was diagnosed in a 58-year-old man. Platelet count was markedly increased and essential thrombocythemia was also diagnosed. Because of LMT disease, coronary artery bypass grafting (CABG) was performed prior to medication for essential thrombocythemia. There were no complications during the operation or in the early postoperative period. Melphalan was administered postoperatively resulting in the decrease of platelet count. Postoperative coronary angiography demonstrated that both grafts were patent; however, immediately after coronary angiography, the patient suffered from a sudden onset of myocardial infarction and cerebral infarction. The therapeutic problems associated with hematological disorder in such patients are discussed in this report. PMID- 8242915 TI - Sir Brian Barratt-Boyes. PMID- 8242916 TI - Biocompatibility of polyethylene and host response to loosening of cementless total hip replacement. AB - A human lymphocyte culture protocol was used to identify the biocompatibility pattern of fine particulate ultra-high molecular weight polyethylene. Polyethylene did not cause an increase in lymphocyte DNA synthesis as assessed by the 3H-thymidine incorporation method on culture Days 1, 3, nor 5. As analyzed with monoclonal activation markers, the polyethylene dependent expression of major histocompatibility complex (MHC) class II antigen as well as interleukin-2 receptor (CD25) was virtually nonexistent. An apparent increase in the amount of CD11b positive monocytes/macrophages from 7% +/- 2% to 22% +/- 6% was recorded. Samples of pseudocapsules of the totally replaced hips (THR) obtained at revision operations for aseptic loosening of cementless prostheses with polyethylene lining of the acetabular component, were obtained from ten patients for immunopathologic studies. In seven cases the prostheses consisted of chromium cobalt-molybdenum steel alloy and in three cases of titanium. Revisions were performed on average 4.6 years (range, 2.5-7) after insertion of the prostheses. The predominant cell in the lining cell layer of the periprosthetic cavity was in each case the CD11b, CD68, and nonspecific esterase positive but endogenous peroxidase-negative macrophage. Proline 4-hydroxylase positive fibroblasts dominated the stroma that was also inhabited by usually perivascular mononuclear cell infiltrations of mainly CD11b/CD68 phenotype with occasional CD4-positive cells. Only few mononuclear cells were activated CD25 positive T cells or CD19 positive B-lymphocytes. In titanium-based THRs, the cytoplasm of the macrophages contained a large number of small metallic particles, although this phenomenon was not seen in chromium-cobalt-molybdenum steel-based THRs. Fine particulate ultra-high molecular weight polyethylene is immunologically relatively insert. Nevertheless, it causes a clear foreign body type of phenomenon in vitro. The loosening of cementless acetabular components was associated with CD11b- and CD68 positive macrophage reaction in the pseudocapsular tissue. In any case, there is no clinical or experimental evidence to suggest that the use of cementless THR prostheses with polyethylene sockets would prevent an adverse biologic host response. PMID- 8242917 TI - Spondylodesis augmented by transpedicular fixation in the treatment of olisthetic and degenerative conditions of the lumbar spine. AB - Fifty-one patients having lumbar or lumbosacral posterolateral fusion with pedicle screw internal fixation were reviewed retrospectively two years after the fusion by an independent observer. In 44 patients, the underlying condition was lytic or degenerative spondylolisthesis. The internal device was removed one year after the fusion. The fusion rate was 94%. The subjective disability was assessed using the Oswestry disability index. The mean Oswestry score was 38% preoperatively, and 24% two years after the fusion. Preoperatively, only every fifth patient was able to work; two years postoperatively, almost 60% held jobs. Complications due to internal fixation were recorded in 57% of the cases. Most of these, such as screw breakage and screw loosening in the sacrum, were of no clinical importance. The use of internal fixation seems to enhance the fusion rate but is associated with more complications than posterolateral fusion without implants. PMID- 8242918 TI - Chromosome study of 249 patients examined for a bone tumor. AB - The authors have performed cytogenetic studies of bone tumors since 1987, and this article, based on previous work, summarizes their experiences in this field. Altogether 304 samples from 249 consecutive patients examined for a possible bone tumor at the Helsinki University Central Hospital, between October 1987 and April 1992, have been analyzed. The series consisted of 86 nonneoplastic disorders, 108 benign and 78 malignant primary bone tumors, and 32 other bone malignancies. In the group of nonneoplastic disorders, one sample from an infectious lesion demonstrated a simple clonal chromosome aberration. Simple clonal aberrations were seen in six of 75 successfully cultured benign tumors. Complex clonal aberrations occurred in 21 of 54 successfully cultured malignant primary bone tumors and in eight of 21 secondary bone malignancies. The complexity of clonal aberrations correlated with the grade of malignancy as the osteosarcomas and chondrosarcomas of high grade demonstrated chaotic abnormalities. The chaotic nature of the aberrations implicates the evolution of several different clones and thus advanced malignant transformation. Six of eight successfully cultured Ewing's sarcomas demonstrated the diagnostic t(11;22)(q24;q12); this translocation was also seen in one primitive neuroectodermal tumor. Cytogenetic study provides increasing knowledge of the biology of bone tumors and is a valuable adjunct in their evaluation. PMID- 8242919 TI - Adductor magnus tenodesis for patellar dislocation. Technique and preliminary results. AB - Fourteen patients with acute or chronic patellar instability had treatment with a new reconstructive procedure. It consists of isokinetic augmentation of the medial patellofemoral ligament (PFL) by making a tenodesis of the distal adductor magnus tendon to the medial border of patella. The patients were reviewed at a mean of 6.9 +/- 0.5 years after operation. In 12 patients, the subjective result was good. One patient had redislocation. PMID- 8242920 TI - Use of the Ilizarov technique after a free microvascular muscle flap transplantation in massive trauma of the lower leg. AB - The use of microvascular flaps has become routine in treating compound tibial fractures with extensive soft-tissue loss. Vascularized bone grafts, or shortening of the leg and later elongation with the Ilizarov technique, have been advocated if the injury involves major bone loss. The authors treated four of these massive injuries by debridement and external fixation with the leg at its normal length. Acute free flap reconstructions (four latissimus dorsi, and one case added with a vascularized iliac crest graft) were performed. Tibial corticotomy was later made above or below the defect (varying from 2.5 to 7 cm) and the bone was corrected with slow (0.5-0.75 mm/day) segmental distraction. This was combined with elongation of the leg in two cases. The consolidation time after corticotomy ranged from nine to 12 weeks. The healing time of the fracture after the initial injury was 12, 14, 22, and 36 months, respectively. In massive injury of the lower leg, maintain the initial length of the leg, and perform early free muscle flap reconstruction and segmental bone distraction under the free flap. PMID- 8242921 TI - Suprascapular nerve entrapment. AB - Fifty-four patients with suprascapular nerve entrapment were evaluated on an average of 5.6 years (range, 2-9 years) after surgical release. There were 32 males and 22 females. Supposed causative factor was exertion at work or vacation in 36 cases. Sixteen patients had an atrophy of the supraspinatus and 26 of the infraspinatus muscle. Conduction time to the supraspinatus muscle was 4.5 ms (range, 2.2-14.4 ms), and to the infraspinatus, 8.6 ms (range, 2.5-43.6 ms). The mean time from the onset of the symptoms to surgery was 2.8 years (range, three months to 14 years). The mean age at operation was 38.4 years (range, ten to 61 years). Two patients were operated on bilaterally within two and four years. All but two patients were operated on at the suprascapular notch. A new cranial approach is advocated. The most dramatic effect of the operation was prompt disappearance of the pain in 24 cases and marked diminishing in 15 cases (72%). At the follow-up evaluation, a moderate atrophy of the supraspinatus muscle was found only in one patient but that of the infraspinatus in 11 patients. There were ten poor long-term results, some of them presumably operated on after wrong diagnoses and some at a wrong region. PMID- 8242922 TI - Gene expression at graft-host interfaces of cortical bone allografts and autografts. AB - The healing processes of autogenous and frozen allogeneic cortical bone grafts were compared in rats. Two unicortical defects created in the proximal metaphysis of the tibia were autografted or allografted with blocks of cortical bone. The biomechanical properties of the graft-host junction and the time-related changes of the expression of genes coding for Type I, II, III, and X collagens and osteonectin were determined at one, two, four, and eight weeks. The strength of the allograft-host bone union was lower than that of autografts at the early stage of incorporation but the difference diminished by eight weeks. Northern analysis of graft mRNAs demonstrated a strong expression of Type I collagen and osteonectin genes at the beginning of autograft incorporation. The allografts showed a lower expression of the corresponding genes at one and two weeks, but the difference diminished thereafter. The gene coding for Type III collagen showed a temporary expression at two weeks both in allografts and autografts, probably because of the formation of inflammatory fibrovascular stroma provided by the host bone at the graft-host interface. No expression of cartilage-specific Type II and Type X collagens were observed, demonstrating that the healing in both grafts proceeded through primary bone healing. Compared with cortical cortical junctions of autografts, the graft-host interfaces of allografts exhibited a reduced osteoinductive process and a slower increase of union strength. PMID- 8242923 TI - The integrity of intervertebral disks after operative treatment of thoracolumbar fractures. AB - This study presents the results of magnetic resonance imaging (MRI) follow-up studies on 14 patients with unstable fractures of T12 or L1. Internal fixation was performed with Harrington rods in eight patients and with transpedicular fixation in six patients. Sixteen intervertebral disks adjacent to the fractured vertebral body in 11 patients showed decreased signal intensity in T2-weighted images. None of the uninjured intervertebral disks showed any detectable signal decrease in T2-weighted images. Temporary fixation of several healthy motion segments of thoracolumbar spine, as is necessary with Harrington instrumentation for 11 months, on average, does not seem to result in an intervertebral disk degeneration detectable on MRI in the patients within a follow-up period of two years. PMID- 8242924 TI - Operative treatment of adolescent idiopathic thoracic scoliosis. Harrington-DTT versus Cotrel-Dubousset instrumentation. AB - Radiographic outcome and complications of Harrington-DTT (H) and Cotrel-Dubousset (CD) instrumentation of idiopathic adolescent thoracic scoliosis were compared retrospectively. The patient material consists of 55 consecutive patients in the H group and 52 consecutive patients in the CD group. The mean age at operation was 15 +/- 2 years (range, 11-19 years) in both groups. The follow-up interval was 43 +/- 18 months (range, 17-91 months) in the H group and 28 +/- 11 months (range, ten to 53 months) in the CD group (p = 0.0001). The preoperative radiographic measurements (Cobb angle of primary and secondary curve, apical rotation, thoracic kyphosis, lumbar lordosis) were fully comparable in both groups. The mean correction of the primary curve at the follow-up evaluation was 47% in both groups. Apical rotation of the primary curve remained almost unchanged in both groups. Rotation of the secondary curve increased significantly in the CD group. Thoracic kyphosis was unchanged in the CD group but decreased in the H group. Spinal balance worsened in 29% of the H cases and in 44% of the CD cases. There were no neurologic complications in either of the groups. Intraoperative lamina fractures (four cases) and postoperative hook dislodgement (five cases) occurred only in the CD group. One distraction rod breakage occurred in the H group. Three reoperations were performed in the H group, nine in the CD group. PMID- 8242925 TI - The cluster phenomenon in patients who have multiple vertebral compression fractures. AB - Little is known about the natural history of patients who have multiple compression fractures. During an eight-year period, eight patients who suffered relentless nonmechanical back pain after an initial compression fracture were evaluated. Radiographs and radionuclide bone scans were obtained in all eight patients and showed the temporal clustering of at least five spontaneous thoracic or lumbar vertebral compression fractures within a period of eight months. The patient database during this eight-year period included 21 patients with a similar number of fractures but without the phenomenon of temporal clustering. Age and gender distribution, premorbid activity levels, presence of secondary causes of osteopenia, dietary history, and vertebral bone density measurements were similar in both groups. Five of the eight patients with cluster fractures were on chronic glucocorticoid therapy for treatment of an underlying illness. All five patients experienced an exacerbation of their chronic illness that required an increase in the dose of their steroid medication within three months preceding the onset of the cluster fractures. No patient in the noncluster fracture control group experienced a similar exacerbation of an underlying illness or required a similar boost in their steroid medication. The temporal clustering of numerous fractures (cluster phenomenon) is a cause of transient severe disability in patients who have profound osteopenia, and may be precipitated by exacerbation of an intercurrent illness or by an associated obligate increase in glucocorticoid medication in a patient with preexisting severe osteopenia. PMID- 8242926 TI - Evaluation of uncemented total hip arthroplasty in patients with avascular necrosis of the femoral head. AB - Thirty-seven uncemented primary porous coated anatomic (PCA) total hip arthroplasties (THA) that were performed in 33 patients for the diagnosis of avascular necrosis (AVN) between June 1984 and December 1986 were reexamined at four- to six-year follow-up periods. Final clinical evaluation showed an 86-point average Harris hip rating (range, 47-100 points), which had improved from an average preoperative rating of 42 points (range, 20-61 points). Thigh pain, which was usually not functionally limiting, occurred in 11 patients (25%), and seven patients (16%) had a moderate or severe limp. Nine patients (20%) required the use of a cane. Long-term radiographic evaluation in 37 hips demonstrated acetabular cup migration in one hip, acetabular lucencies in four hips (11%), and acetabular bead loosening in ten hips (27%). Femoral subsidence occurred in five hips (14%) and femoral bead loosening in 11 hips (30%). Thirty hips (81%) demonstrated radiographic evidence of stable bony ingrowth. No revisions were performed for aseptic loosening, but two prostheses were removed for infection. These results of uncemented THA in patients with AVN appear to be as good or better than the results reported for cemented arthroplasty in AVN at this interval of a follow-up period. This study supports the use of uncemented devices, especially with the newer designs and increasing surgical experience in this difficult patient population. PMID- 8242927 TI - Prospective preoperative evaluation of anterior cruciate ligament instability of the knee joint and results of reconstruction with patellar ligament. AB - Of 54 patients with chronic (n = 49) or acute (n = 5) anterior cruciate ligament instability, 52 were evaluated after mean 16 months' follow-up interval after open (n = 18) or arthroscopic assisted (n = 36) bone-tendon-bone patellar ligament reconstruction. Preoperative Knee Signature System side-to-side difference in anterior displacement decreased from 7.9 mm to 3.5 min at follow-up evaluation (p < 0.0001). An objectively satisfactory limit of 5 mm in side-to side difference was achieved in 73% of the patients. At follow-up evaluation, there was a positive pivot shift sign in eight knees (one definite and seven trace). Average Lysholm knee scores improved from 69 to 83. The only significant difference between the arthroscopic assisted and open groups was smaller side-to side anterior displacement difference in the arthroscopic group (2.2 mm versus 4.8 mm, p = 0.002). Results suggest that more accurate and isometric placement of the graft is possible with the arthroscopic-assisted technique. PMID- 8242928 TI - Case report of a successful closed reduction without anesthesia. AB - Intraarticular dislocation of the patella is a rare clinical entity. Horizontal dislocation occurs when the patella rotates 90 degrees about its horizontal axis. Of the 33 previously reported cases, only six have been successfully reduced by closed manipulation. Furthermore, all previously successful closed reductions have required general anesthesia to accomplish the reduction. Intraarticular dislocation of the patella in a 24-year-old man was successfully reduced in a closed fashion without anesthesia. PMID- 8242929 TI - Proprioception after knee arthroplasty. The influence of prosthetic design. AB - It is recognized that awareness of joint position in the knee deteriorates because of aging, anterior cruciate injury, or osteoarthrosis. Proprioception was studied in osteoarthritic and prosthetic knees, including both posterior cruciate ligament (PCL)-retaining and PCL-sacrificing designs. An improvement in proprioception was observed after knee arthroplasty. A PCL-retaining prosthesis was found to confer a greater improvement than a PCL-sacrificing design. PMID- 8242930 TI - Expandable endoprosthetic reconstruction of the skeletally immature after malignant bone tumor resection. AB - The mainstay of local control of primary bone malignancies in the skeletally immature has been amputation or, in selected cases, rotationplasty. The development of expandable endoprostheses has permitted an alternative approach for local control in the growing child. Between January 1985 and December 1987, 12 skeletally immature patients with primary malignant bone tumors were treated with extremity reconstruction with cemented custom-expandable endoprostheses after wide resection of their lesions. All patients were observed until death (four) or revision (two) with a minimum two-year follow-up period for the survivors (average, 3.1 years). Seven patients have undergone a total of 11 expansions and one patient was lengthened with a revision-expandable prosthesis. Four patients have not needed expansion. Eight patients have had a total of ten complications. Seven of the ten complications (70%) were prosthesis related and associated with failure of the expansion mechanism. The Musculoskeletal Tumor Society (MSTS) overall rating was good to excellent in seven patients (58%), fair in three (25%), and poor in two (17%). In five distal femoral arthroplasties and one total femoral arthroplasty where the tibial bearing component was cemented through the physis, tibial and epiphyseal growth was observed to be normal and equal to the nonoperative side. This suggests that partial central epiphyseal and physeal ablation does not cause physeal arrest. Although the high rate of expansion mechanism failure necessitates redesign, preliminary results suggest that expandable endoprostheses do offer an alternative to amputation and rotationplasty as a means of local control and extremity reconstruction in children with primary malignant bone tumors. PMID- 8242931 TI - The growth of orthopedics in Finland. PMID- 8242932 TI - Allograft transplantation in aggressive or malignant bone tumors. AB - Twenty-three patients with aggressive benign or malignant bone tumors had segmental allograft replacements after resection of the tumors. The mean age of the patients was 30 years of age. The mean length of the allografts was 14 cm. The allografts consisted of intercalary allograft (5), allograft arthrodesis (12), and allograft-prosthetic composite (6). Intramedullary fixation with or without additional compression plate was used to osteosynthesize all the osteotomy sites with the exception of one in which a long nail plate was used. A supplement of autogeneic bone grafts at the allograft-host junction was performed in 18 patients. Seven patients received preoperative and postoperative chemotherapy for high grade sarcomas. At an average follow-up period of four years, 78% of the patients had excellent or good results. The union rate of the grafts was 86%. Supplement with autografts shortened the union time of the allograft from 15 to eight months. One patient had a tumor recurrence and one died of the disease. Other complications included one delayed union, two nonunions of the allograft, two late infections, and one neuropathy. Allograft transplantation is an acceptable method for tumor reconstruction. The adjuvant chemotherapy did not have significant adverse effects on its outcome. Complications that relate to allografting are common, but most of them are salvageable. PMID- 8242933 TI - The effect of staple size, orientation, and number on torsional fracture fixation stability. AB - Staples have been used for fixation of metaphyseal fractures, but there are no guidelines for placing the staples to ensure maximum fixation stability. This study investigated the effect of staple size, orientation, and number on torsional stability in simulated transverse fractures. Six homogeneous foam cylinders were cut transversely, stapled back together using a power driver, and rotated coaxially on a servohydraulic test system. Staples with bridge widths of 7, 10, 13, and 16 mm, and leg lengths of 7, 10, 15, and 20 mm were used. Each test was performed seven times. Torsional holding power was increased when two staples were oriented in opposite directions (45 degrees) and had bridges wide enough to maintain bone purchase with respect to the fracture line. Fixation effectiveness increased with the number of staples: the use of two staples (as opposed to one) yielded the biggest increase; further significant increases were also observed for the use of three, four, and five staples. Bridge width and leg length had minimal effect, as long as bone purchase sufficient to avoid cutout was maintained from the staple to the fracture surface. PMID- 8242934 TI - Penetration of the distal femur by the distal locking device of Brooker Wills interlocking nail. AB - Of seven patients with complications caused by penetration of the distal locking device through the distal femur, three had penetration of the distal locking device into the knee joint. Two patients had penetration into the patellofemoral joint. One patient had penetration into the lateral cortex of the distal femur and another patient had penetration into the medial cortex of the distal femur. All but one patient required removal of the rods and/or the distal locking device. The main causes of complications include rod rotation, previous surgery, preexisting deformity of the femur, and failure to insert the rod into its proper location distally. PMID- 8242935 TI - Radial head fracture. A potentially complex injury. AB - A prospective study was undertaken on 50 acute, consecutive fractures of the radial head. Clinical examination and stress testing were performed on each patient. A valgus stress radiograph, in conjunction with clinical stress testing, was used to assess competence of the medial elbow ligaments. Axial stability of the radius was determined by the axial stress test. Eleven patients (22%) sustained a comminuted radial head fracture; all had instability either to valgus or axial stress. Seventeen patients (34%) had a displaced vertical shear type fracture or an impacted fracture of the radial neck. All sustained some injury to the medial collateral ligament, with variable degrees of valgus elbow instability. Twenty-two (44%) of the fractures were minimally or nondisplaced. All 22 were stable. Radial head fracture morphology can be used to assess the likelihood of associated soft-tissue injury. PMID- 8242936 TI - Regulation of extracellular matrix genes during fracture healing in mice. AB - The expression of several collagens, cartilage proteoglycan core protein (aggrecan), and osteonectin was investigated during healing of standardized fractures of the mouse tibia. Total RNA was extracted from fracture calluses and normal bone, and analyzed by Northern hybridization using specific cDNA probes. Based on the predominant fibrillar collagen type expressed, three phases of fracture healing were observed. Type III collagen expression characteristic for the inflammatory stage increased first, followed by Type II collagen expression indicative of the cartilaginous reparative phase, and by Type I collagen production, characteristic for the ossification and remodelling stages. Type IX collagen and aggrecan expression coincided with Type II collagen expression during the chondrogenesis. The mRNA for Type X collagen, a marker for hypertrophic chondrocytes during endochondral ossification, occurred somewhat later than that of other cartilage-specific genes. Osteonectin mRNA was present throughout the healing process and peaked during rapid new bone growth. This study confirms that the pattern of gene expression in the murine system is similar to, but somewhat faster than, other species analyzed (rat, rabbit, human). PMID- 8242937 TI - Supracondylar fractures of the humerus in children. Technique and results of crossed percutaneous K-wire fixation. AB - Displaced extension type supracondylar fractures of the humerus are associated with neurovascular injuries and malunion (cubitus varus). Reduction is not usually difficult during the first few hours, but it is a problem to maintain. After reduction, the shoulder is placed in abduction external-rotation with the elbow flexed under the C-arm. This position locks the proximal segment of the fracture, and the posteromedially rotated distal segment can then be brought back into an anatomic position under the "medial pillar." In this position, an excellent lateral view is obtained and the "fish tail" deformation can be corrected. Crossed percutaneous K-wires are introduced to maintain reduction. An anteroposterior radiograph can then be freely obtained, and there is no need for hyperflexion at the elbow. Twenty-six children were treated between 1987 and 1992. Excellent results were obtained. PMID- 8242938 TI - Chronic septic arthritis caused by Borrelia burgdorferi. AB - Chronic arthritis occurs in 10% of Lyme disease patients. A patient had chronic septic Lyme arthritis of the knee for seven years despite multiple antibiotic trials and multiple arthroscopic and open synovectomies. Spirochetes were documented in synovium and synovial fluid (SF). Polymerase chain reaction (PCR) analysis of the SF was consistent with Borrelia infection. Persistent infection should be excluded with silver stains and cultures in any patient with chronic monoarticular arthritis and a history of Lyme disease. PMID- 8242939 TI - Intraoperative glove perforation. A comparative analysis. AB - A prospective study was conducted to determine the rate of puncture of the inner glove when various double-gloving protocols were used for elective total joint procedures. Group 1 consisted of 22 cases in which two pairs of regular latex gloves were used. Group 2 consisted of 25 cases in which outer "orthopaedic" gloves were used over regular latex gloves. Group 3 consisted of 24 cases in which Repel gloves were used between two regular latex gloves. The patients were randomly assigned to one of the three groups. All gloves, both inner and outer, were tested for holes at the end of the procedure. All gloves that were changed for presumed intraoperative holes were also tested. The type and duration of the procedure was also documented. Of the inner holes, there were significant differences between each group. Group 1 (double latex) and Group 2 (outer orthopaedic/inner regular) had significantly more holes than Group 3 (Repel between two regular gloves). There were no holes in Group 3, and Group 2 offered significant protection over Group 1. PMID- 8242940 TI - Bilateral predominant radial nerve crutch palsy. A case report. AB - Axillary crutch palsy, although well described in the literature, is rarely seen in clinical practice. Little has been written in the contemporary orthopaedic literature. The purpose of this case report is to report on a well-documented axillary crutch palsy with bilateral, predominantly radial, nerve dysfunction that resolved spontaneously in eight weeks. PMID- 8242941 TI - Knee immobilization inhibits biomechanical maturation of the rabbit medial collateral ligament. AB - In this investigation, the effect of immobilization on the structural biomechanical properties of the immature rabbit medial collateral ligament (MCL) was quantified. The right hindlimbs of ten, three-month-old female New Zealand White rabbits were immobilized. The hindlimbs of the first group (n = 5) were immobilized for one month, until the rabbits were four months of age. The hindlimbs of the second group (n = 5) were immobilized for three months, until the rabbits were six months of age. Left hindlimbs were not surgically treated and served as contralateral controls. Fifteen normal animals were kept in similar cages until they were killed in subgroups at three, four, or six months of age. Biomechanical investigations after the animals were killed included the testing of a number of MCL structural properties, including prefailure and strength tests. Results showed that the maturation of MCL structural properties were inhibited during immobilization with signs of structural degradation becoming more obvious at the longer period of immobilization. Results also demonstrated that the structural mechanical properties of the contralateral MCLs were significantly altered. These results suggest that knee mobility is essential to the normal maturation of MCL mechanical properties and that contralateral MCLs cannot be used as normal controls in this immature rabbit model of immobility. PMID- 8242942 TI - High-purity resident tissue macrophage isolates from human synovium and periprosthetic tissues using immunomagnetic techniques. AB - Destruction of periarticular and periprosthetic bone by activated macrophages, a process often termed "macrophage mediated osteolysis," is recognized as a leading mechanism of aseptic arthroplasty failure. To develop effective interventional approaches and increase the longevity of implanted joint prostheses, the pathobiology of activated human-synovium-derived macrophages needs to be better characterized. The first step toward achieving this research objective is the acquisition of pure populations of macrophages from human synovial tissue. A simple, fast, and highly efficient method for isolating a relatively pure population of macrophages from periprosthetic tissue received from either primary or secondary arthroplasty is presented. This technique uses murine monoclonal antibodies (IgG) that recognize a phagocyte-specific marker, CD68, for primary binding, and sheep anti-murine IgG antibodies bound to polystyrene-coated magnetic microspheres for secondary binding. While the primary antibody specifically labels CD68-positive phagocytes in the digestion of synovial and periprosthetic tissue, the secondary antibody bound to polystyrene-coated iron oxide beads facilitates the removal of CD68-positive cells from CD68-negative cells by anchoring the former with a magnet. This protocol requires centrifugation only in the washing steps, which reduces the frequency of cell death and altered cell morphology. The patient population includes three primary and eight revision arthroplasties. The tissue macrophage isolation protocol yielded on average 4 x 10(5) cells/g tissue, of which 91% were viable nonspecific esterase positive macrophages. The experimental results suggest that immunomagnetic beads coupled to anti-CD68 enable the isolation of a purified population of resident tissue macrophages suitable for further biologic characterization. PMID- 8242943 TI - Atraumatic spontaneous hemarthrosis associated with Lyme arthritis. A case report. AB - A 20-year-old male athlete had Lyme arthritis and an associated atraumatic spontaneous hemarthrosis of the knee. Lyme arthritis is a common and well documented manifestation of Lyme disease, but an association with an atraumatic hemarthrosis to date has not been reported. Clinical diagnosis was confirmed by serologic testing. Treatment consisted of ceftriaxone, 1-g intravenous for 14 days. PMID- 8242944 TI - Human immunodeficiency virus transmission between surgeons and patients in orthopaedic surgery. AB - Surgeons are at risk for contracting human immunodeficiency virus (HIV) from patients. The risk of HIV transmission between patient and surgeon and the potential strategies to reduce the risk of HIV transmission are important. Continuous occupational exposure makes the risk of HIV transmission greater for surgeons than patients. Although the risk of seroconversion after a single exposure is relatively low, the risk for surgeons is more appropriately expressed as a cumulative lifetime risk. The estimated cumulative risk of HIV seroconversion for surgeons may be as high as 1 to 4%. Currently available strategies to prevent HIV transmission require knowledge of the mechanisms of exposure. Adequate barriers, such as double-gloving, waterproof outerwear, and face protection, should be worn for most, if not all, orthopaedic procedures. Additional specific strategies, however, are required to minimize sharp injuries. Surgeons should report any significant exposure to the occupational health department of their institution. Hospitals should have appropriate guidelines and procedures for counseling exposed surgeons, HIV testing of source patients, consideration of zidovudine prophylaxis, and disability insurance policies for surgeons who are occupationally infected with HIV. PMID- 8242945 TI - A new approach to the internal fixation of unstable pelvic fractures. AB - Eighteen patients with horizontally or vertically unstable pelvic fractures were operated on with a new intrapelvic technique. There were 11 horizontally unstable and seven vertically unstable pelvic ring fractures. The fractures were exposed through a low Pfannenstiel or lower midline incision combined with an incision at the iliac crest. This is referred to as the ilioanterior approach. At the operation, all lesions in the symphysis, rami, and in the sacroiliac region were reduced and fixed with reconstruction plates and screws. No intraoperative complications ensued. The average intraoperative blood loss was 930 ml and the operation time averaged two and one-half hours. The obtained reduction was maintained in all but one sacral fracture. All fractures united. The functional recovery was uneventful in all the patients. The good results obtained in this relatively small series encourage further development of operative techniques in unstable pelvic fractures. PMID- 8242946 TI - Partially purified reindeer (Rangifer tarandus) bone morphogenetic protein has a high bone-forming activity compared with some other artiodactyls. AB - Noncollagenous proteins, including bone morphogenetic protein (BMP), were extracted in 4 mol/l guanidinium hydrochloride (GuHCl) from the pulverized and HCl-demineralized matrix of reindeer, bovine, sheep and porcine bone. To remove water-soluble material, the GuHCl solution was dialyzed against water and water insoluble material and redissolved in 4 mol/l GuHCl. Gelatin peptides were removed by extraction in 0.25 mol/l citrate buffer (pH 3.1). The yield consisted mostly of large complexes and protein molecules of molecular weight less than 35,000 daltons. Isoelectric focusing of the material showed three to four different protein molecules: three acidic and one neutral. Bone-forming activity was investigated by implanting 0.6-15.0 mg of partially purified protein preparation into the thigh muscles of BALB mice. Radiologically detectable formation of new bone required 0.6 mg of reindeer BMP, 2.5 mg of bovine BMP, 5.1 mg of sheep BMP, and 8.0 mg of porcine BMP. A rough estimate of the area of the deposits showed that reindeer BMP had the highest bone formation activity, and porcine had the lowest. The formation of new bone was confirmed histologically. It is suggested that the differences in osteogenic activity are due to quantitative differences in BMP constituents or in the degree of complex formation in the protein preparations. Also immune and other defense mechanisms may generate differences in osteogenic response. PMID- 8242947 TI - Distraction bone healing. AB - Bone formation by distraction was studied using three different experimental models: (1) Physeal distraction of the sheep radius was performed in 20 animals. (2) Distraction after osteotomy of the radius was carried out in 39 sheep. (3) Mandibular distraction after osteotomy was performed in 17 sheep. Formation of the organic matrix and osteogenesis were studied by radiographic, histologic, and biochemical methods as well as by electron microscopy. The mode of osteogenesis was essentially similar in all of these distraction models. Bone formation was preceded by organization of the collagenous matrix in the distraction area. In the beginning of the distraction, the gap was composed of a heterogeneous cell population, with large polymorphic fibroblast-like cells. The cells in the central part differentiated into fibroblasts, which remained functionally active as long as distraction proceeded. During physeal distraction, bone formed from the epiphyseal and metaphyseal sides as well as from the surrounding perichondrium. Also, in osteotomy distraction of both tubular bone and mandible, bone formed centripetally from the osteotomized bone ends toward the center of the gap. The organic matrix was composed almost solely of Type I collagen in the earliest stages, suggesting that the mode of osteogenesis differs from bone repair by fracture callus. The structure of the distracted segment was mainly lamellar trabecular. Corticalization of the lengthened bone segment occurred gradually after several months. PMID- 8242948 TI - Partial closure of the epiphyseal plate. Principles of treatment. 1978. PMID- 8242949 TI - Early muscle changes after immobilization. An experimental study on muscle damage. AB - Immobilization of the rabbit knee in extension has previously been shown to damage the vastus intermedius profundus (VIP) muscle. To examine the mechanism of the early stages of the muscle damage, the authors studied creatine kinase activity in serum, and both light and electron microscopic changes in the affected muscle. The right knee was immobilized in an extended position using a splint, and thigh muscles were removed at various intervals, up to 48 hours after immobilization. The left hindlimb served as a control. Creatine kinase levels in serum rose ten hours after the onset of the immobilization. The enzyme levels reached a substantial peak by 24 hours, and plateaued thereafter. Light microscopic changes were not observed within 48 hours, but in electron microscopy distinct mitochondrial swelling and crystal abnormalities were seen as early as ten hours. The ultrastructural changes of mitochondria remained constant for up to 36 hours and decreased thereafter. At 48 hours of immobilization, also myofibrillar disorganization was seen. It appears that immobilization of the rabbit knee in extension rapidly leads to signs of remarkable damage to the VIP muscle. These suggest leakage of the cell membrane and metabolic disturbances. The ultrastructural changes observed share common features with muscle damage caused by ischemia, uncoupling agents, and inherited mitochondrial myopathies. PMID- 8242950 TI - Specific collagen mRNAs elucidate the histogenetic relationship between the growth plate, the tissue in the ossification groove of Ranvier, and the cambium layer of the adjacent periosteum. A preliminary report. AB - In situ hybridization was employed to study cellular phenotypes in the ossification groove of Ranvier in young rabbits. The expression of Type II collagen mRNA was seen in chondrocytes in all layers of the growth plate. Messenger RNA for the cartilage-specific Type II collagen was also detected in cells of the inner layer in the ossification groove and of the cambium layer of the adjacent periosteum, providing strong evidence for the views of Ranvier that these cells originate in the growth plate. These findings modify the interpretation of the pathogenesis of diseases of the growing skeleton. PMID- 8242951 TI - The effect of immobilization on the torsional strength of the rat tibia. AB - Torsion testing is considered a suitable method for measuring the strength of long bones. The aim of the current study was to investigate the effect of immobilization on the mechanical strength of rat bone and its relation to mineral content. The influence of freezing on the bone strength was also studied. The precision of the method, studied by comparing the maximum torque capacity of the contralateral tibiae of intact rats, was found to be 6.4%. No significant dominance of either side in the strength of the tibia could be observed. Tibiae that had been frozen for one week before thawing had a significantly lower maximum torque capacity and rigidity than fresh contralateral bones, the differences being -6.3 +/- 3.2% (mean +/- SEM) (p < 0.05) and -9.37 +/- 9.48% (p < 0.05), respectively. The change in the maximum angle of deformation was not statistically significant. Three weeks' cast immobilization reduced the femur ash weight by 13.8 +/- 2.5% (p < 0.001) relative to that of the nontreated contralateral leg. The maximum torque capacity and rigidity of the tibia also decreased in the immobilized leg, the differences being 11.6 +/- 3.5% (p < 0.01) and 16.6 +/- 5.3% (p < 0.01), respectively. The maximum angle of deformation was not significantly influenced by immobilization. The maximum torque capacity of the tibia correlated well with femur ash weight (r = 0.71, p < 0.001). The method used here is valid for measuring the strength of diaphyseal bones; it showed weakening of the bone during three weeks' cast immobilization. PMID- 8242952 TI - Ultrasonographic evaluation of patellar tracking in children. AB - Patellar positions with the knee in extension and in 10 degrees and 20 degrees flexion were measured by ultrasonography in both knees of 20 children with patellar dislocation and in both knees of 30 healthy children for controls. Ultrasonography proved to be applicable to measuring the distance between the most anterior point of lateral femoral condyle and patellar apex, the sulcus angle and the vertical position of the patella. Compared with normal knees, the sulcus angle was wider and the patellar position more lateral and cranial in both the dislocated and asymptomatic knees of the patients. These results suggest that false tracking of the patella in early flexion is a predisposing factor to patellar dislocation. PMID- 8242953 TI - Isthmic spondylolisthesis in symptomatic and asymptomatic subjects, epidemiology, and natural history with special reference to disk abnormality and mode of treatment. AB - Observations during the past 35 years on operative and conservative treatment of adult and young patients with isthmic spondylolisthesis, demonstrate the main indications of fusion are pain and risk of further progression of the slip. At one time, posterior fusion was recommended. Posterolateral fusion in situ was adopted in 1975, and during the past ten years anterior fusion combined with reduction and transpedicular fixation has been performed in severe slips. Surgery was mainly indicated to alleviate pain or to prevent further progression of the slip. It seems possible to alter the natural course of the disease by operation, but it is difficult to prove the connection between the radiographic findings and the pain. The epidemiologic study shows that the prevalence of spondylolisthesis in Finland is 6%, but there are no significant differences between these subjects and controls without spondylolisthesis. Symptomatic patients may have pain, however, even after a long observation time. Prevention of the slip is difficult because, statistically, 90% of the slip has already occurred when the patient is first seen. Posterolateral fusion in situ is the method of choice, especially for most young patients. Anterior fusion is indicated if the slip is greater than 40% 50%. Reduction of the slip is indicated in exceptional cases only. PMID- 8242954 TI - Proliferation of the hypertrophic chondrocytes of the growth plate after physeal distraction. An experimental study in rabbits. AB - Reports on the effects of physeal distraction on chondrocyte proliferation are controversial, and the biologic mechanism of lengthening is still open. Proliferation of the growth plate chondrocytes was studied after gradual physeal distraction in 12 growing rabbits. Distraction of the distal femoral physis was performed with an external fixation device for three to 14 days. Proliferating cells were labeled with 5-bromo-2'-deoxyuridine one hour before killing the animal, and labeled chondrocytes were localized with a specific monoclonal antibody. Labeled chondrocytes were observed in the hypertrophic zone after distraction of three to seven days, in addition to labeling in the proliferative zone; in the remaining physes, the distribution of labeled cells was normal. Physeal widening occurred in all distracted specimens, and a fracture-separation, usually through the hypertrophic zone, in all but one. The labeled hypertrophic cells were seen on the epiphyseal side of the separation gap, five to 20 cells in each specimen. Although hypertrophic chondrocytes are known to be metabolically active, proliferation of these cells has never been reported before. Hypertrophic chondrocytes seem capable of proliferation under special circumstances. PMID- 8242955 TI - Cementless total hip arthroplasty for congenitally dislocated or dysplastic hips. Technique for replacement with a straight femoral component. AB - A new straight cementless stem was used for replacement of 67 dislocated or severely dysplastic hips. There were 45 hips with complete dislocation, 27 of which were cases after Schanz osteotomy. Technical solutions for various deformities are presented here. The acetabular component was placed at the level of the original cotyloid cavity or some lower position. In hips after total dislocation a metaphyseal shortening osteotomy was combined with distal advancement of the greater trochanter with intact attachment of the abductor muscles. This method was appropriate also for the femora, where high-seated Schanz osteotomy was previously performed. If the diaphysis was too narrow for the stem, it was split about 10 cm both anteriorly and posteriorly. In cases with unilateral total dislocation where Schanz osteotomy had been seated low, metaphyseal segmental shortening with angular correction was performed and the stem was used as an intramedullary nail. Special attention was paid to achieve sufficient abduction strength to balance the pelvis and abolish Trendelenburg limp and to restore leg length. The clinical and radiographic results of the consecutive series were assessed three to five years after the arthroplasty. Pain relief and the functional results including improvement of gait were generally good, primary complications were few, but the loosening and revision rate of the smooth-threaded acetabular component was unacceptably high. There were no problems with the press-fit cups. In general the outcomes were good even when reoperation was required. PMID- 8242956 TI - Prognosis after conservative and operative treatment in Perthes' disease. AB - The prognosis of Perthes' disease was studied after femoral intertrochanteric varus osteotomy in 112 patients (126 hips) and after conservative noncontainment treatment (Thomas splint) in 96 patients (106 hips). The radiographic results at or near skeletal maturity after containment treatment were good in 45%, fair in 21%, and poor in 34% of the hips. The figures after noncontainment treatment were 21%, 18% and 61%, respectively. The results support the idea that containment of the femoral head in the acetabulum should form the basis of treatment for Perthes' disease. It was very difficult to delineate signs for good prognosis; age at onset of the disease and a good containment three months postoperatively in the operatively treated group were the only signs with prognostic significance. In both groups, one fifth of the patients had an extremely poor outcome. Treatment other than varus osteotomy or noncontainment treatment should be considered in these patients. PMID- 8242957 TI - Ceramic endoprosthesis in total hip arthroplasty. AB - Aluminia-on-aluminia hip prosthesis with titanium alloy stem was used in 255 cases, for 143 patients with cemented acetabular cup and 112 with an uncemented screw cup. The average age of the patients was 62 years. Of the patients, 35 were treated bilaterally. The indication for surgery was osteoarthritis in 186 cases, fractures and nonunions of the hip joint in 34 cases, and congenital dislocations of the hip joint in 16 cases. Previous surgery of the hip joint was recorded for 43 cases. All procedures were primary arthroplasties. Only personal clinical examinations together with radiographic studies were accepted as clinical data. Three patients failed to show up for routine follow-up evaluation and these were excluded from the series. Revision operation was classified as failure. The indications for revision were aseptic loosening, late infection, and fracture of the acrylic cement resulting in loosening of the acetabular or femoral component. The acetabular component was cemented in 143 patients and the mean follow-up period for these patients was 6.7 years (range, 1-12 years). In this series, a revision operation was undertaken for 16 patients (11%). In 12 cases, broken acrylic cement resulted in acetabular aseptic loosening. In the series of 112 patients with uncemented titanium screw cup, the mean follow-up period was 3.6 years (range, 1-7 years) and a revision operation was undertaken in seven cases (7%). In two, the indication was late infection; in one, technical failure; and in two, progression of Pigmented villonodular synovitis. Acetabular aseptic loosening resulted in revision in only two cases. With cementless acetabular component bone transplantation is indicated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242958 TI - Nonspecific diskitis in children. A nonmicrobial disease? AB - Eighteen patients were treated for nonspecific diskitis (diskitis without microbial cause) at the University Children's Hospital in Helsinki during 1970 1990. The mean age at admission was three years three months. The characteristic findings were restriction of spinal mobility and an elevated erythrocyte sedimentation rate (ESR). Radiographic narrowing of the affected disk space was seen four to five weeks after the initial symptoms. An operative biopsy was made in 16 patients, the histologic diagnosis being chronic or subacute nonspecific inflammation in ten, other nonspecific changes in two, and normal tissue in five. Treatment was either bed rest and antimicrobial medication, or bed rest alone. After a follow-up period of five months to nine years, all patients were clinically normal. All but two had radiographic signs of healed disk space defects. Nonspecific diskitis in children is an entity with unknown cause. The clinical picture is typical, but diagnostic difficulties may be encountered during the first weeks when radiographic signs are absent. The disease is self limiting in most cases, and only minor radiographic changes remain after healing. The results after routine operative biopsy raise the question of a possible nonmicrobial cause. PMID- 8242959 TI - Willem Anton den Hartog Jager (1913-1993). PMID- 8242960 TI - Rhabdomyolysis: a review of the literature. AB - Rhabdomyolysis (lysis of skeletal muscle cells) is a potentially lethal syndrome with a broad spectrum of clinical and biochemical findings. Myalgia, pigmenturia and elevated activity of serum creatine kinase are the common features. Fulminant rhabdomyolysis may be associated with severe metabolic disturbances and involvement of other organ systems. Cardiac arrest, compartment syndrome and acute renal failure are the major complications. The extent of the life threatening complications of rhabdomyolysis strongly depends on early diagnosis and adequate therapy. As the repair mechanism of striated muscle functions very well, the prognosis of adequately treated rhabdomyolysis is excellent. This article reviews the present state of knowledge of clinical and biochemical diagnosis of rhabdomyolysis, the pathophysiologic background, the classification and the etiological provocative factors. PMID- 8242961 TI - Cerebral arteriovenous malformations in The Netherlands Antilles. High prevalence of hereditary hemorrhagic telangiectasia-related single and multiple cerebral arteriovenous malformations. AB - Seventeen patients with symptomatic cerebral arteriovenous malformations (AVMs) were diagnosed between 1980 and 1990 in the Leeward Islands of the Netherlands Antilles. Five patients had multiple AVMs. The annual incidence of symptomatic AVMs was 1.1/100,000. The mean age of presentation was 35 years. In 6 patients cerebral AVMs were associated with hereditary hemorrhagic telangiectasia (HHT); 4 of these patients had multiple AVMs. We conclude that HHT is frequently encountered in Netherlands Antillians with symptomatic and multiple cerebral AVMs. PMID- 8242962 TI - Brain infarct causes under the age of fifty: a comparison between an east-Asian (Thai) and a western (Dutch) hospital series. AB - Proportion and causes of brain infarction among young persons may differ between Western world and developing countries, which may require different diagnostic and treatment strategies. We compared a Thai series of 56 hospitalized brain infarct patients younger than 50 years (16% of 357 similar patients from all ages) with a series of 55 Dutch patients (8% of 694 patients). The Thai patients had 12% (95% confidence interval: 3-21) more often a cardioembolic stroke cause, due to a higher frequency of rheumatic heart disease in the Thai series. The percentage of patients with atherothrombotic stroke cause was higher in the Dutch series, whereas the percentage of those with hematologic abnormalities, or those with vasculopathies did not differ. Patients in the Thai series had 17% (95% confidence interval: 7-27) more often an unidentified stroke cause, which was probably related to less extensive ancillary investigations compared with the Dutch series. We conclude that especially centers in developing countries that treat young stroke patients should be supplied with ancillary testing facilities, such as cardiac echo, carotid ultrasound, and some laboratory facilities. Early treatment of infections, especially acute rheumatic fever, could lower the incidence of stroke in the young. PMID- 8242963 TI - Natural history and risk factors of unruptured cerebral aneurysms. AB - We report the results of a statistical analysis of the long-term outcome of 54 patients with 72 unruptured cerebral aneurysms, and identify the factors for predicting subsequent ruptures. Twenty-two patients died during the observation period, which averaged 43.7 months. The 5-year survival rate was 56%. Aneurysms ruptured in 11 patients (20.4%), 10 of whom died without undergoing surgery. The annual bleeding rate was 1.92%. The average size of the 11 ruptured aneurysms was 13.1 mm. In 4 patients, however, bleeding occurred in unruptured cerebral aneurysms of 4 and 5 mm, which suggests that leaving unruptured cerebral aneurysms of less than 10 mm in size untreated is hazardous. According to the Cox proportional hazards model, the shape and location of the aneurysm and the presence of hypertension were the most important factors for predicting a subsequent rupture. Our data suggest that unruptured cerebral aneurysms arising from the vertebrobasilar and middle cerebral arteries of 10-19 mm size and of multilobes had a statistically high probability of subsequent bleeding. Although 20 patients with 28 unruptured cerebral aneurysms were followed through repeated examinations, we could not correlate the risk of subsequent bleeding with changes in the size of the aneurysm. PMID- 8242964 TI - The influence of a standard meal on Sinemet CR absorption in patients with Parkinson's disease. AB - We studied the influence of dietary protein intake on the plasma level profile of levodopa, carbidopa, and 3-O-methyldopa and clinical efficacy in 12 patients with idiopathic Parkinson's disease after intake of one levodopa-carbidopa 200/50 controlled release tablet (Sinemet CR; LC-CR). The tablet was given 1 h before the protein rich meal on one day (fasted) and together with the meal on an other day (non-fasted). Higher levodopa and carbidopa concentrations were reached when the LC-CR was taken 1 h before the meal, but the plasma level profile for levodopa was flatter in the non-fasted state. The area under the curve for levodopa was slightly higher in the fasted condition. For the clinical variables walking and tapping slightly better clinical results (P = 0.08) were found in the fasted condition with the higher levodopa levels. If the patient on levodopa is in a clinically satisfactory condition, then non-fasted condition could be preferred because of the smooth plasma level profile demonstrated. However, if the initial levodopa concentrations are not in the critical range to be effective for the patient, the advice should be to take the drug in a fasted condition. PMID- 8242965 TI - Circadian blood pressure patterns in four cases with hemodynamic brain infarction and prolonged blood-brain barrier disturbance. AB - We report on four patients with hemodynamic brain infarction and pathological circadian blood pressure patterns with nocturnal hypotension which gave rise to a prolonged disturbance of the blood-brain barrier. Besides at least one severe stenosis of the internal carotid artery, there was an untreated chronic arterial hypertension and a pathologically reduced vasomotor reactivity after CO2 stimulation in all patients. The 24-h blood pressure monitoring then carried out showed a distinctly pathological circadian profile with hypertensive day values and nocturnal hypotension with minimum values of 95/50 mm Hg. The range of variation between day and night values was significantly raised (systolic: 20% +/ 2.15%; diastolic: 22.9% +/- 2.58%) compared to patients with essential hypertension as well as normotensive subjects (P < 0.01), and was in excess of 40% in the individual case. There was a slow recovery of the blood-brain barrier after drug-induced normalization of the pathological circadian blood pressure profile. We conclude that the registration of circadian blood pressure patterns may be of prognostic and therapeutic relevance. It may also contribute to further clarification of the pathophysiological significance of blood pressure variability for the development of brain infarction. PMID- 8242966 TI - A double-blind study of the efficacy of apomorphine and its assessment in 'off' periods in Parkinson's disease. AB - Five patients with idiopathic Parkinson's disease with severe response fluctuations were selected for a randomized double-blind placebo-controlled study, concerning the clinical effects of subcutaneous apomorphine and its assessment in 'off'-periods. The study was designed as five n = 1 studies, in which every patient was his own control. The effect of apomorphine was studied by using the Columbia rating scale and quantitative assessments, using tapping, walking and pinboard. There was a significant positive effect of apomorphine, in a mean optimal dose of 2.7 mg, with a mean latency of onset of 7.3 min and a mean duration of response of 96 min. After pretreatment with domperidone, no significant adverse effects were observed. Tapping showed the highest correlation with rigidity and bradykinesia. Walking showed a high correlation with stability and gait. Pinboard testing did not give additional information. The first conclusion was that apomorphine proved to be a significantly effective dopamine agonist, proven now also by a double blind placebo-controlled study. Secondly it was concluded that assessment of clinical effect in parkinsonian patients can be performed best by combining the Columbia item tremor with tapping and walking scores. PMID- 8242967 TI - Rotatory seizures of frontal lobe origin. AB - We report a patient with true rotatory seizures. The CT scan showed a right frontal calcified lesion and the EEG showed focal epileptiform discharges over the right frontal region. The seizures responded well to phenytoin therapy. PMID- 8242968 TI - Longstanding isolated cerebral systemic lupus erythematosus in an 8-year-old black girl. Resemblance with multiple sclerosis. AB - We describe a 12-year-old girl with SLE who presented with longstanding isolated neurological symptoms resembling MS. The literature concerning the difficulties in diagnostic procedures in cerebral SLE are reviewed in comparison to MS. PMID- 8242969 TI - Primary ectopic intramuscular meningioma of the thigh. AB - A case of a primary ectopic intramuscular meningioma of the thigh in a 20-year old male patient is presented. The possible histogenesis of primary ectopic meningiomas is summarized. PMID- 8242970 TI - Successful treatment of amoebic meningoencephalitis in a Chinese living in Hong Kong. AB - Primary amoebic meningoencephalitis due to Naegleria fowleri was found in a 38 year-old Chinese man living in Hong Kong who presumably acquired the infection from swimming in a hot spring in neighbouring China. Amoebic cysts were identified in tissue taken from a brain abscess. The patient responded to surgical drainage and a 6-week course of amphotericin B, rifampicin and chloramphenicol. This is one of 6 cases of successful treatment of primary amoebic meningoencephalitis documented in the medical literature. PMID- 8242971 TI - Meningeal carcinomatosis as presenting symptom of a gallbladder carcinoma. AB - Meningeal carcinomatosis as presenting symptom is rare. We report a patient who presented with an isolated abducens nerve palsy and an unsteady gait. At autopsy the primary tumor proved to be a gallbladder carcinoma. Only one similar case has been reported. Several aspects are discussed, including the route by which the tumor cells could have reached the subarachnoid space. PMID- 8242972 TI - A unique case of intracranial metastasis from lung carcinoma. AB - We describe a case of an intracranial metastasis from a lung carcinoma in the same location from which an intracranial meningioma had been completely removed surgically eleven months earlier. It is demonstrated that the metastasis originated in the scar of the first operation. PMID- 8242973 TI - Three-phase bone scintigraphy. Asymmetric patterns in the upper extremities of asymptomatic normals and reflex sympathetic dystrophy patients. AB - Three-phase Tc-99m MDP scans of 61 patients with asymptomatic upper extremities randomly mixed with 17 studies of patients previously diagnosed with reflex sympathetic dystrophy were blindly interpreted by three observers. Asymmetry in any of the phases was recorded and a final diagnostic impression made. Thirteen of 17 reflux sympathetic dystrophy studies were rated abnormal by at least two observers. Mild to striking asymmetry was occasionally seen in all three phases in asymptomatic upper extremities. Twenty of 61 asymptomatic patients (33%) were rated abnormal by at least one observer, and 5 of 61 studies (8%) were rated abnormal by all observers. Asymmetries in normal patients occurred more commonly in the earlier phases, while asymmetry in the delayed images was mild in all but one. Tightening the criteria to exclude mild asymmetry in delayed images resulted in unacceptably low sensitivity for reflex sympathetic dystrophy (29%). Interobserver variability was most prominent in the flow and immediate images. In the diagnosis of reflex sympathetic dystrophy a greater reliance should be placed on the delayed images, which in themselves have an overall sensitivity of 94%. It is important, however, to recognize that occasional mild and rare moderate asymmetries in even the delayed images of normal individuals result in an overall lower specificity of 77%. PMID- 8242974 TI - Intraosseous fat necrosis and infarction associated with pancreatitis. AB - A case of trauma-induced pancreatitis with subsequent intraosseous fat necrosis and infarction is presented. The young patient demonstrated multiple cortical lytic lesions of the lower extremities and bilateral sterile joint effusions. Several radiographic modalities were employed in the workup of his disease. Related findings and pathophysiologic considerations are reviewed. PMID- 8242975 TI - Scintigraphic evaluation of multifocal hemangioendothelioma of bone. AB - Multifocal bone involvement was seen on bone scintigraphy in a young woman being examined for a suspected bone lesion noted on pelvic radiographs. Biopsy of the third lumbar vertebral body demonstrated a vascular lesion. The tumor was resected and a bone graft was performed. The patient was followed with serial scans to assess the viability of the bone graft and course of the disease. This report analyzes the scintigraphic characteristics of this rare tumor and emphasizes the importance of bone scintigraphy in the diagnosis, treatment, and follow up of this condition. PMID- 8242976 TI - Myocardial SPECT and left ventricular performance study using a single Tc-99m teboroxime injection. Comparison with thallium-201 myocardial SPECT. AB - Myocardial SPECT using teboroxime was compared to thallium SPECT in 26 patients undergoing cardiac catheterization. Agreement between thallium SPECT and teboroxime SPECT for the identification of myocardial segments was 209/235 (89%). A significant correlation was found in 15 patients between the left ventricular ejection fraction by teboroxime first-pass study and the ejection fraction by contrast ventriculography. In nine patients with myocardial infarction and/or multivessel coronary artery disease, the ejection fraction revealed a mean decrease from 0.52 at rest to 0.46 at exercise. Teboroxime makes it possible to perform an exercise first-pass study of left ventricular ejection fraction followed by myocardial perfusion imaging. PMID- 8242977 TI - Radionuclide simultaneous dual-isotope stress myocardial perfusion study using the "three window technique". AB - Several methods are available for performing radionuclide stress myocardial perfusion studies. All of these methods require two separate acquisition and processing iterations, which is time-consuming and inconvenient for the patient. The authors introduce a new method using the "three window technique" that they developed to perform simultaneous dual-isotope imaging for stress myocardial perfusion studies. In addition to Tl-201 and Tc-99m windows centered at 70 KeV and 140 KeV, respectively, a third window centered at 105 KeV, representing scattered radioactivity from Tc-99m, is established. By subtracting the radioactivity in the third window from the radioactivity in the Tl-201 window, "crosstalk" interference is significantly reduced. The simultaneous dual-isotope imaging method acquires and processes resting and stress myocardial perfusion images simultaneously. It halves imaging time and doubles patient throughput, improves scheduling flexibility, and reduces patient waiting time and discomfort. PMID- 8242978 TI - Temporal changes in myocardial uptake on serial bone scans in two cases of presumed senile cardiac amyloidosis. AB - Two unusual examples of cardiac uptake in elderly patients who had received serial bone scans, one with a "waxing" and the other with a "waning" pattern of myocardial tracer uptake, are presented. An apparent temporal discordance between echocardiographic and scintigraphic findings is noted. Neither of the two patients had clinical, electrocardiographic, or other laboratory evidence of old or new myocardial infarction. Possible causes for the temporal changes are described. Presumptive diagnoses of senile cardiac amyloidosis were made based on clinical findings, noninvasive imaging, and comparison with a historic cohort at the Veterans Administration Medical Center in Houston. PMID- 8242979 TI - The rim sign in acute cholecystitis. Comparison of radionuclide, surgical, and pathologic findings. AB - Nonvisualization of the gallbladder associated with the rim sign of enhanced pericholecystic hepatic activity has been established as a useful and highly specific sign of acute cholecystitis. This study's purpose was to determine the pathophysiologic cause of the rim sign. Thirty-seven surgery-bound patients with a clinical diagnosis of acute cholecystitis were studied. A group of 20 patients with a definite rim sign were compared to a group of 17 patients without a rim sign; all with nonvisualization of the gallbladder. Radionuclide, surgical, and pathologic findings were correlated. In patients with the rim sign and in those without the rim sign, microscopic review of all surgical specimens revealed transmural reaction of the gallbladder wall in 95% and 59% of cases, respectively. The surgeon found inflammation extending beyond the gallbladder wall in 75% of rim sign cases, but in only 35% of case without the rim sign. Liver tissue was attached to the gallbladder specimen in three cases, and the findings support the association of hepatic inflammation with the presence of a rim sign. Pathologic correlation and surgical evidence suggest that the rim sign is caused by the spread of inflammation through the gallbladder wall and into adjacent liver tissue. Transmural reaction is required before the inflammatory process can reach the liver. A high-count, high-intensity technique seems to best demonstrate the rim sign. Once demonstrated, further delayed imaging is unnecessary. PMID- 8242980 TI - The diagnostic value of grading hyperperfusion and the rim sign in cholescintigraphy. AB - Biliary scans of 84 hospitalized patients believed to likely have acute cholecystitis, including 55 scans that had a radionuclide angiography phase, were retrospectively evaluated to determine the frequency of the rim sign and hyperperfusion, and to test the hypothesis that more intense hyperperfusion or rim sign is associated with a greater severity of gallbladder pathology ("complicated" acute cholecystitis). In 65 of the 84 cases there was surgical intervention (including 43 from the 55 cases whose scans had a radionuclide angiography phase). "Complicated" acute cholecystitis was considered present if there was gangrene, perforation, empyema, necrosis, ulceration, or fibrous exudation. Each scan was evaluated for the presence of a rim sign and arterial hyperperfusion to the region of the gallbladder fossa. The intensities of these secondary signs of acute cholecystitis were then graded as "mild" or "marked." Subdividing the rim sign and hyperperfusion into a "marked" category considerably improved the specificity, positive predictive value, and likelihood ratio (positive) for the diagnosis of acute cholecystitis, but even more so for the complicated subgroup when marked hyperperfusion or marked rim sign were the criteria used for a positive study. Approximately 50% of the patients with acute cholecystitis had hyperperfusion and a rim sign, and approximately 15% had marked hyperperfusion and a marked rim sign. Of the patients with acute cholecystitis, the only ones with marked hyperperfusion or a marked rim sign were those who had complicated acute cholecystitis. The data demonstrate an association between greater intensity of the rim sign or hyperperfusion and greater severity of gallbladder pathology in patients with acute cholecystitis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8242981 TI - Postoperative diuresis renography and ultrasound in patients undergoing pyeloplasty. Predictors of surgical outcome. AB - The authors reviewed 50 pediatric patients (56 renal units) who underwent pyeloplasty and had serial preoperative and postoperative diuresis renogram and ultrasound studies. Of those patients that clinically improved with surgery, 73% and 91% showed improved renographic patterns in postoperative studies at 3 and 12 months, respectively. Ultrasound demonstrated more gradual improvement in grade and pelvic diameters over longer study intervals. Diuresis renography is an excellent predictor of surgical outcome within 3 months of pyeloplasty and showed change well in advance of that seen on ultrasound. Sonography is better suited to longer term evaluation of grade, pelvic diameter, and renal size. PMID- 8242982 TI - A comparison of methods in measurement of effective renal plasma flow and glomerular filtration rate in clinical practice. AB - Using a single injection clearance method as a reference test, 24-hour creatinine clearance and gamma camera estimation of ERPF and GFR were compared in a sample of the Japanese adult patient population. The reference tests included ERPF with I-131 OIH and GFR with In-111 DTPA, single blood sample, and 1-hour external counting over the heart for each tracer. The 24-hour creatinine clearance and gamma camera methods were equally accurate in estimating GFR or ERPF compared with the clearance methods. The gamma camera methods were less accurate for GFR than ERPF. Among many gamma camera estimation formulas for GFR, the renal Tc-99m DTPA uptake best correlated with the reference GFR (compared to Schlegel's GFR or Gates' GFR). The authors found that the simpler the procedure and data processing, the better the results. Origin of errors in estimating ERPF and GFR, the procedures of choice, patient population studied, quality control achievable in clinical practice, and the clinical situations the clinicians are likely to encounter are discussed. PMID- 8242983 TI - Scintigraphic findings of renal oncocytoma. AB - Tc-99m DTPA scintigraphy of most renal tumors characteristically discloses a vascular mass, without retention of activity on serial images. Two cases, encountered during hypertensive work-up, are presented with vascular renal masses and persistent DTPA uptake. Oncocytoma was confirmed pathologically in both cases. The authors conclude that renal oncocytomas have a relatively distinctive scintigraphic appearance, which may permit accurate preoperative diagnosis. PMID- 8242984 TI - Tc-99m sestamibi uptake by cerebellar metastasis from bronchogenic carcinoma. AB - Tc-99m sestamibi has been used to detect primary brain and lung tumors. The authors report a patient who underwent brain imaging to differentiate tumor from abscess in the cerebellum because of a ring-like enhancement lesion on a brain CT scan. An abnormal area of increased sestamibi uptake in the right cerebellum was demonstrated planar and on a SPECT imaging. The removed tumor was confirmed to be poorly differentiated metastatic carcinoma. PMID- 8242985 TI - The role of morphine-augmented cholescintigraphy in the detection of acute cholecystitis. PMID- 8242986 TI - Effect of gravity in preventing a potential false-positive lung scan. PMID- 8242987 TI - Arm splints seen on bone scan in a volleyball player. PMID- 8242988 TI - Unusual scintigraphic pattern for postsurgical bile leak. PMID- 8242989 TI - Gallium-67 scan of a patient with African variety of Burkitt's lymphoma involving Waldeyer's ring. PMID- 8242990 TI - Hepatic uptake of Tc-99m MDP in ischemic hepatitis. PMID- 8242991 TI - Retained esophageal activity on iodine-131 survey in patient with benign esophageal stricture. PMID- 8242992 TI - Hydrocele and epididymitis mimicking testicular torsion on scrotal scintigraphy. PMID- 8242993 TI - Autonomously functioning thyroid rests following total thyroidectomy for Graves' disease. PMID- 8242994 TI - Bridging rib fractures. Scintigraphic and radiographic findings. PMID- 8242995 TI - Is there a role for corticosteroid prophylaxis in patients at increased risk of adverse reactions to intravascular contrast agents? PMID- 8242996 TI - The use of the Wallstent in aorto-iliac vascular disease. AB - Forty-three patients had self-expanding endovascular Wallstents inserted for aorto-iliac arterial disease. A total of 59 Wallstents were inserted into 50 iliac arteries. There were 27 males and 16 females with a mean age of 62.51 years. Included in the series were 19 iliac occlusions, 21 severe atheromatous disease of the aortoiliac segment, two severe dissections following conventional angioplasty and one re-stenosis of a previously dilated iliac segment. All of the stents were successfully deployed but there were five significant complications. One case of distal embolism was treated by surgical embolectomy but the other four were treated by an additional percutaneous technique. At follow-up after 6 months all patients remain symptomatically improved. Aorto-iliac stenting is a cost-effective treatment in selected cases of severe aorto-iliac disease. PMID- 8242997 TI - Surgical clip artefact mimicking arterial stenosis: a problem with magnetic resonance angiography. AB - In three patients undergoing three-dimensional phase contrast magnetic resonance angiography (MRA) for evaluation of renal transplant arteries, a convincing stenosis was demonstrated that was not present on conventional angiograms. In each patient there was a stainless steel surgical clip in the soft tissues adjacent to the renal artery. Although the clip artefact was easily recognized on the acquired tomographic magnetic resonance slices, it was completely indistinguishable from a true stenosis on the final projection images. This may limit the use of MRA in post-surgical vascular evaluation unless review is made of the phase contrast images. PMID- 8242998 TI - Radiologically-guided cutting needle biopsy for suspected malignancy in childhood. AB - Twenty-seven cutting needle biopsies were performed on 25 children with suspected malignancy using computed tomographic (CT, 22) or ultrasound (US, 5) guidance. Anatomical sites were: retroperitoneum 6, liver 4, kidney 4, abdomen/pelvis 4, thorax 4, bowel 2, neck 1. Sixteen patients (64%) underwent subsequent open biopsy (5), marrow biopsy (2) or resection (9). There was complete concordance between the histological findings from the open or marrow biopsy and the previous needle biopsy in 12 of these 16 patients; in two patients the needle biopsy was misleading, causing inappropriate initial treatment in one. In two other patients needle biopsy was correct but lacked specific diagnostic features. Needle biopsies were performed under general, local or Ketamine anaesthesia. There were no apparent complications related to these procedures. We believe that radiologically-guided cutting needle biopsy should replace open biopsy in most children with solid malignant lesions. It can easily be performed during a single anaesthetic episode which allows radiological evaluation, biopsy, bone marrow and cerebrospinal fluid sampling. However, the potential for sampling error and histological variation within these tumours needs to be borne in mind. PMID- 8242999 TI - Endosonographic monitoring of transurethral cryoprostatectomy. AB - Cryoprostatectomy, the use of subzero temperatures to ablate prostate tissue gained favour in the 1960s because of its speed and lack of haemodynamic effects. It fell from use because the freezing process could not be monitored and this led to a high incidence of local complications. We have performed transrectal real time ultrasound in 12 patients undergoing cryoprostatectomy. In all the freezing process was easily visualized as an advancing hyperechoic 'iceball' with distal acoustic shadowing. Monitoring allowed maximum prostate ablation without breaching the prostatic capsule. There were no significant complications and all but one patient gained symptomatic relief. Ultrasound at 1 month showed a heterogeneous echo pattern with very little cavity formation but by 3 months a significant cavity was seen. This study demonstrates that it is possible to monitor the freezing process during transurethral cryoprostatectomy. This, theoretically, should significantly reduce the local complications encountered in previous studies. PMID- 8243000 TI - Polymyositis: MRI-appearance at 1.5 T and correlation to clinical findings. AB - The thigh regions of 10 patients suffering from polymyositis were examined by magnetic resonance imaging (MRI) at 1.5 T; follow-up examinations were performed in all patients. MRI findings were correlated with findings at physical examination and serological parameters. In four patients focal areas of hyperintensity on T2-weighted (T2W) spin echo (SE) images were found within muscles. They were not detectable on T1-weighted (T1W) images. In three of these cases enhancement within the musculature was seen following i.v. administration of Gd-DTPA on T1W images. In another five patients there was varied replacement of muscle by fatty tissue. In one patient the muscle appeared completely normal. On follow-up three patients showed definite changes in imaging findings; two correlated well with clinical and biochemical findings while the third exhibited massive deterioration clinically with mild improvement on imaging. Seven patients exhibited no change on MRI; four of these had shown a significant clinical change in the course of the disease, whilst three were clinically stable. In our small series MRI using SE technique (T1W and T2W) and contrast enhancement to date seems to be of limited value for the assessment of muscular changes during therapy. The i.v. use of Gd-DTPA did not add to MRI's diagnostic value in this series. PMID- 8243001 TI - Detection of pneumothorax with lateral shoot-through digital radiography. AB - The aim of this study was to compare the performance of digital frontal radiographs vs digital lateral shoot-through radiographs for the diagnosis of pneumothoraces in supine patients. A total of 146 pairs of radiographs were performed on 32 ventilated patients on the Intensive Care Unit. Each radiograph was independently assessed by three observers for the presence or absence of a pneumothorax. A degree of confidence was assigned to each observation and an image quality score was given to each radiograph. At least two out of three observers positively diagnosed a pneumothorax in 13/146 (8.9%) of the frontal radiographs compared to 43/146 (29.4%) of the lateral radiographs (P < 0.0001), but suboptimal images were obtained more often with lateral shoot-through radiographs than with frontal radiographs. We conclude that digital lateral shoot through radiographs are significantly more sensitive than digital frontal radiographs for the diagnosis of pneumothoraces in supine patients. PMID- 8243002 TI - Radiographic patterns of intrathoracic disease in breast carcinoma: prognostic implications. AB - The chest radiographic appearances of patients with intrathoracic metastases from breast cancer are variable and their relationship to patient outcome is unknown. This study aimed to classify and determine the frequency of various radiographic patterns and, in addition, to establish whether these patterns can be used to predict patient prognosis. A retrospective study was made of the patients of a major breast unit who over a 3 year period were found to have developed intrathoracic metastases. In each case, the earliest plain chest X-ray showing signs of metastasis was reviewed and the frequency of the different radiographic patterns determined. Comparison was made between these patterns and patient mortality. The chest radiographs of 92 patients who developed intrathoracic metastases during a 3 year period were analysed. Pulmonary nodules were found in 66%, pleural effusions in 41% (bilateral in 18%), lymphadenopathy in 25%, lymphangitis in 18% and pleurally-based nodules in 11%. Median survival of the whole group was 13.5 months. Patients with bilateral, but not unilateral, effusions had a significantly poorer prognosis (median survival 3 months). Although the median survival of patients with lymphangitis seemed poor (5.5 months), this difference was not statistically significant. Multivariate analysis showed bilateral effusions to be the only independent predictor of worse outcome. We have found that of the plain radiographic patterns seen in intrathoracic metastasis from breast cancer, only bilateral pleural effusions can be used to predict a worse prognosis than that associated with intrathoracic metastasis as a whole. PMID- 8243003 TI - Free intra-peritoneal gall-stones following laparoscopic cholecystectomy. AB - Laparoscopic cholecystectomy is now the operation of choice for symptomatic gall bladder stones. Two cases of retained free intra-peritoneal gall-stones following this procedure are described; the first resulting in a peritoneal granuloma and causing initial confusion in diagnosis, and the second seen incidentally in a biloma. In the appropriate clinical setting free intra-peritoneal gall-stones must be added to the differential diagnosis of abdominal and pelvic calcification. PMID- 8243004 TI - Image processing of videofluoroscopy of patients with velopharyngeal insufficiency and hypernasal speech. AB - Eleven patients with hypernasal speech and velopharyngeal insufficiency (VPI), together with five normals, were evaluated by videofluoroscopy to assess velopharyngeal movement, velar lift and lateral pharyngeal wall movement. Computer processing of the images obtained was used to compensate for initial poor quality images by contrast and edge enhancement techniques and to provide objective measurement of the movements involved. It was demonstrated that objective computer aided analysis of videofluoroscopic images is feasible and may provide additional subtle diagnostic information when nasendoscopy is unavailable. In addition, results obtained showed an increased degree of velar lift and lateral pharyngeal wall movement for the more severely affected patients. These suggest a compensatory mechanism in operation for the more severe cases of VPI. PMID- 8243005 TI - Antenatal detection of congenital diaphragmatic hernias: the northern region experience. AB - This study analyses 7 years of data relating to congenital diaphragmatic herniation in the Northern Region. Ninety-nine confirmed cases were notified to the Northern Region Fetal Abnormality Survey between 1985 and 1991, which is an incidence of 0.35 per 1000 births. A specific diagnosis of CDH was made by antenatal ultrasound in 21 fetuses. The rate of detection improved from 6% to 41% but this had no impact on survival; severe pulmonary hypoplasia was found in the majority dying in the first 48 h. Another structural abnormality occurred in 29% and was the primary antenatal scan diagnosis in 11 fetuses. No other reliable indicator of outcome was identified, including the gestation when detected. The incidence of chromosomal abnormality was 5% and Fryns' syndrome 4%. PMID- 8243006 TI - Iodixanol and ioxaglate in cardioangiography: a double-blind randomized phase III study. AB - A new non-ionic, dimeric contrast medium iodixanol (Nycomed AS, Norway) has a very low osmolality and is isotonic with blood. It has been compared with ioxaglate (Hexabrix, Laboratoire Guerbet, France) in a double-blind, randomized, parallel trial. The aims of the trial were to evaluate and compare the safety (vital signs, adverse events, discomfort and clinical-chemical parameters in blood and urine) and radiographic efficacy (diagnostic information and radiographic density) of iodixanol 320 mg I/ml vs ioxaglate (Hexabrix 320 mg I/ml) in coronary angiography and left ventriculography. Seventy-six patients referred for cardioangiography, two patients were withdrawn, 36 receiving iodixanol and 38 ioxaglate were included in the trial. Six patients (16%) in the iodixanol group and 16 (42%) patients in the ioxaglate group reported adverse events (P = 0.02). One serious adverse event occurred in the iodixanol group where a patient experienced transient cortical blindness and transitory global amnesia, but the patient recovered completely the day after the examination. Twenty-six patients reported injection-associated sensation of warmth in the iodixanol group versus 34 in the ioxaglate group (P = 0.06). Following contrast injection there were no differences between the groups regarding vital signs (ECG, heart rate, left ventricular pressures). Both contrast media were well tolerated by the kidneys, and on average only minor effects on clinical-chemical parameters in blood and urine were observed in the two groups. The radiographic efficacy was good in both groups. PMID- 8243007 TI - Urethral steinstrasse--clinical experience and radiographic findings. AB - Urethral steinstrasse is rare and requires immediate intervention. The radiographic and cystoscopic findings in two patients with urethral steinstrasse are described. PMID- 8243008 TI - The use of effervescent agents in the small bowel meal examination. AB - Some perceived disadvantages of the small bowel meal examination are failure of adequate distension, lack of a double contrast effect and the duration of the procedure. A new use for effervescent granules during the small bowel meal is described which reduces the examination time by 70% and reduces the radiation dose to the patient. PMID- 8243009 TI - Technical report: use of a directional needle for antegrade guide wire placement when performing femoropopliteal angioplasty. PMID- 8243010 TI - Case report: thromboembolism during transfemoral arteriography--radiographic demonstration with histopathological correlation. AB - Thromboembolism is a well described complication of transfemoral arteriography. Its origin and course is rarely demonstrated. We describe a case of thromboembolism with histopathological correlation demonstrating the origin and course during contrast injection. PMID- 8243011 TI - Case report: invasive renal angiomyolipoma--sonographic and CT features. PMID- 8243012 TI - Case report: emphysematous tuberculous pancreatitis diagnosis by ultrasound and computed tomography. AB - We describe a case of simultaneous tuberculous and pyogenic infection of the pancreas in a 58-year-old Asian man who presented with a pyrexia of unknown origin. There was no evidence of disseminated tuberculosis or immuno-deficiency. The diagnosis was confirmed by ultrasound-guided percutaneous pancreatic aspiration and subsequent progress assessed by sequential computed tomography (CT) and ultrasonography. Complete recovery was achieved on anti-tuberculous chemotherapy without surgical intervention. PMID- 8243013 TI - Case report: simple liver cyst masquerading as a gastric leiomyoma--a diagnostic pitfall. AB - A 47-year-old woman presented with abdominal pain. A barium meal and CT scan suggested a gastric leiomyoma. Follow-up CT showed that a cyst of the left lobe of the liver was responsible for the appearances. The differential diagnosis is discussed. PMID- 8243014 TI - Posterior rectocele or perineal herniation--what's in a name? PMID- 8243015 TI - Lower limb colour flow imaging: a comparison with ankle:brachial measurements and angiography. PMID- 8243016 TI - [Preliminary evaluation of an association of polyunsaturated acids in primary hypertriglyceridemia]. AB - The above study was intended to evaluate changes in triglyceride blood level in subjects treated with a polyunsaturated fatty acid combination: docosahexanoic (DHA) and eicosapentaenoic (EPA) acid, compared to changes observed in patients treated with arachidonic acid. The results obtained show that a significant reduction of triglyceride blood level was achieved and was more marked in patients treated with the DHA-EPA combination compared to those treated with arachidonic acid only. An increase in the index of fatty acid unsaturation was observed in all patients without significant differences between treatments. PMID- 8243017 TI - [Cigarette smoking and hormonal changes in the course of the menstrual cycle]. AB - ACTH, cortisol and PRL plasma levels were determined on day 7, 14, and 21 of the menstrual cycle in 20 females with hypertrichosis of whom 10 were smokers and 10 non-smokers. Significantly raised levels of all hormones considered were found in smokers throughout the cycle. These findings suggest a possible relationship between objective symptoms (hypertrichosis) and nicotine, mediated by complex changes in the endocrine pattern and presumably also due to the action of nicotine on the central nervous system. PMID- 8243018 TI - [Evaluation of the effect of enalapril in hypertensive patients with non-insulin dependent diabetes mellitus]. AB - We selected 15 hypertensive non insulin-dependent diabetics in reasonable metabolic control, treated for 6 months with enalapril 20 mg/die. The results showed a significant reduction of mean arterial blood pressure and triglycerides without interference on the metabolic control. PMID- 8243019 TI - [Evoked motor and somatosensory potentials in diabetic neuropathy]. AB - The present study was aimed at evaluating the information obtained from evoked motor and sensitive potentials in a population of non-insulin-dependent diabetics compared to information yielded by electroneurographic study. Besides, we wanted to check any possible correlation between metabolic control and changes of evoked potentials. Electroneurographic study of 20 patients revealed signs of peripheral nervous impairment in 11 of these while results were normal in the remaining 9. Analysis of findings resulting from testing of evoked potentials showed a statistically significant increase of these parameters both in the group of patients with neuropathy and in the controls. Since these methods explore longer stretches of the nervous pathways, they are apt to identify lesions located in areas not accessible to common diagnostic methods. Our data did not show a correlation between metabolic control, as assessed by fructosamine assay, and latency of evoked motor and sensitive potentials, thus indicating that other factors, such as environmental and genetic ones, are apt to influence the onset of diabetic neuropathy. PMID- 8243020 TI - [Cardiovascular effects of amitriptyline in therapeutic dosages. Echocardiographic study]. AB - The authors studied the cardiovascular effects of amitriptyline at therapeutic plasma concentrations in 15 depressed patients (6 M. 9 F.) without cardiovascular disease both before treatment and after six months of therapy. The cardiovascular effects were evaluated by means of electrocardiographic and 2D-echocardiographic examinations in basal conditions and after hand-grip stress test. The effects of isometric hand-grip exercise (IHG) on left ventricular size and performance were studied non invasively in all patients at rest and after 3 min. of IHG at 30% of maximum contraction. Left ventricular internal diameter was measured at end diastole and end-systole on LV echograms, and blood pressure was measured by sphygmomanometer. Our data confirmed the depressant effect of amitriptyline even on healthy myocardium, an effect that becomes manifest only at handgrip stress with a significant reduction of ejection fraction (form 70.6 to 66.4%; p < 0.001), while ECG and arterial blood pressure did not change throughout the study. This goes to show that treatment with tricyclic antidepressants always has a latent depressant effect on myocardial contractility that becomes clinically evident under stress, as well as in subjects with heart disease and in the elderly. Hence the need to monitor left ventricular function, as well as ECG and blood pressure, and to exercise great caution in prescribing tricyclic antidepressants to subjects with a history of myocardial failure. PMID- 8243021 TI - [Acromegalic arthropathy]. AB - Six acromegalic patients, three males (aged 28 to 48 years) and three females (aged 57 to 75 years), with GH-producing pituitary adenoma, were studied through clinical examination, laboratory and instrumental tests. In all the patients frequent involvement of large joints, with crepitus and provoked pain, was found; while articular mobility was normal especially in the dorso-lumbar spine, a frequent seat of pain. Radiology showed typical features of an osteoarthritic process with characteristic widening of articular spaces, especially in weight bearing large joints in symptomatic patients. The evolution of this arthropathy lead to anatomo-clinical pictures almost indistinguishable from osteoarthritis; however, in the early stages, the marked cartilaginous hypertrophy is responsible for peculiar anatomo-radiological pictures, principally represented by widening of articular spaces and intervertebral discal spaces, especially in the dorso lumbar spine. As far as bone metabolism is concerned, neoproduction and reabsorption, both increased, proceed simultaneously; bone mass reduction is described in some segments. In our study, the two patients with active acromegaly showed bone mass reduction in the lumbar spine. PMID- 8243022 TI - [Amlodipine in isolated systolic hypertension in the aged]. AB - The authors studied the efficacy and safety of a long half-life dihydropyridine calcium-antagonist--amlodipine--in a single daily dose in the treatment of isolated systolic hypertension in the elderly. Ninety-seven subjects, age range 65-85 years, with isolated systolic hypertension were randomized into two groups, similar for age, weight and height, of 47 subjects submitted to active therapy and 50 subjects to placebo, respectively. The study was divided into two phases: in the first one, of three weeks, the first group was administered amlodipine at the dose of 5 mg/day; in the second period, which lasted 9 weeks also in case of systolic pressure normalization, amlodipine at the dose of 10 mg/day was administered. Forty-one subjects of the active drug group and 41 of the control group completed the study. The results were determined as mean +/- standard deviation; statistical analysis was completed by the Student T test. In the amlodipine group, at the end of the third week, a significant decrease in systolic arterial pressure was reported; at the end of the follow-up a further decrease in systolic arterial pressure was observed in the active treatment group as compared to the control group (p < 0.001). The other parameters considered (E.C.G., routine haematochemical tests) did not show significant changes in the two groups during the whole study period. Furthermore, during the follow-up, neither orthostatic hypotension nor reflex tachycardia, events which require particular attention during antihypertensive therapy in the elderly, were observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243023 TI - [The role of magnesium in glucose homeostasis: therapeutic implications]. AB - The authors examined the literature on the close relationship of magnesium to carbohydrate metabolism. It was found that a diabetic condition is apt to bring about marked changes in body magnesium content and its compartmental distribution, and that the administration of dietary magnesium supplements can have a favorable influence on metabolic control, thus reducing insulin requirement and counteracting the progression of late diabetic complications. PMID- 8243024 TI - [Computerized tomography and magnetic resonance in the evaluation of neoplastic processes localized on the floor of the third ventricle]. AB - In our experience, neoplastic lesions of the floor of the third ventricle are frequently responsible for neurologic and dysendocrine symptoms. Ct scan and MR imaging are the most reliable techniques of neuroimaging in the diagnostic study of neoplastic masses affecting this region. In this work, CT and MR appearance of lesions of the III ventricle floor are discussed with the aim of contributing to the differential diagnosis of neoplasms and their surgical and medical management. PMID- 8243025 TI - [The role of tomography (X-ray and magnetic resonance) in the diagnosis and determination of the extension of tumors in the roof of the third ventricle]. AB - We describe the CT scan and MR appearance of neoplastic lesions affecting the roof of the third ventricle. The neoplastic lesions of this region may be of neuroepithelial origin (sub-ependymoma, colloid cyst), meningeal cell origin (meningiomas), metastatic or primary localization of lymphoproliferative tumors (lymphomas) and dysembryogenetic origin (epidermoid tumor). CT scan and MR imaging are useful techniques for diagnosis and planning of surgical or medical treatment. The primary role of these imaging techniques, is discussed in the quantification of extension, biologic malignancy and anatomical relationships between pathologic lesion and normal nervous tissue. PMID- 8243026 TI - [Idiopathic dilatational cardiomyopathy. A clinical case]. PMID- 8243027 TI - [Primary fibromyalgia]. PMID- 8243028 TI - Communicative competence and theory of mind in autism: a test of relevance theory. AB - Sperber and Wilson's (1986) relevance theory makes explicit the role of the comprehension of intentions in human communication. Autistic people have been hypothesized to suffer from a specific and characteristic impairment in the ability to attribute such mental states (e.g., beliefs, intentions); a lack of "theory of mind". According to relevance theory, then, autistic people should have specific difficulties with the use of language for communication. Relevance theory allows precise predictions about the levels of communicative competence that should be possible with either no, first-order only, or second-order theory of mind ability. Three experiments are reported which tested predictions following from the analysis of figurative language in terms of relevance and theory of mind, in able autistic and normal young subjects. The results lend support to relevance theory. In addition, the findings suggest that some autistic subjects are eventually able to attribute mental states. Lastly, the results demonstrate close links between social and communicative understanding in autism and normal development. PMID- 8243029 TI - The colour cognition of children. AB - It is well known that young children have great difficulty in stating the appropriate colour name when presented with a colour stimulus, even though they may be effective at matching one colour stimulus with another and possess a good vocabulary of colour terms. The present studies demonstrate a further, but distinct, impairment in dealing with coloured stimuli. Three- to four-year-old children were given the task of distinguishing an appropriately coloured option from one with an inappropriate colour. Despite displaying a preference for the appropriate colour over the inappropriate one, children found the task very difficult. In contrast, they succeeded on the task more frequently when the alternatives were presented verbally, rather than in terms of two pictures. We discuss the results within a theoretical framework which distinguishes the access to object-colour knowledge from colour naming. The model makes divisions within the mental templates required for object matching. In particular, colour detail is functionally separated from shape detail and appears to be represented in more than one code. PMID- 8243030 TI - Behavioral and neuronal changes during treatment of mixed transcortical aphasia: a case study. AB - The development of language mechanisms outside the traditional language regions is evidenced by the case of K.S., a patient with chronic mixed transcortical aphasia, that is, good repetition performance compared to all other language abilities. The aphasia was caused by an infarction of the left a. carotis interna that completely destroyed the left perisylvian language region and adjacent structures. Five years post onset the patient underwent intensive aphasia therapy. In the treatment setting chosen, both speech production and comprehension abilities improved as demonstrated by adequacy and reaction time scores. It is argued that the behavioral changes observed are likely to be due to (a) newly acquired communicative strategies and (b) neuronal changes within K.S.'s brain. The behavioral changes are explained in terms of strengthening of synaptic connections and formation of Hebbian cell assemblies corresponding to words. PMID- 8243031 TI - Darwinian algorithms and the Wason selection task: a factorial analysis of social contract selection task problems. AB - In four experiments with 760 subjects, the present study examined Cosmides' Darwinian algorithm theory of reasoning: specifically, its explanation of facilitation on the Wason selection task. The first experiment replicated Cosmides' finding of facilitation for social contract versions of the selection task, using both her multiple-problem format and a single-problem format. Experiment 2 examined performance on Cosmides' three main social contract problems while manipulating the perspective of the subject and the presence and absence of cost-benefit information. The presence of cost-benefit information improved performance in two of the three problems while the perspective manipulation had no effect. In Experiment 3, the cost-benefit effect was replicated; and performance on one of the three problems was enhanced by the presence of explicit negatives on the NOT-P and NOT-Q cards. Experiment 4 examined the role of the deontic term "must" in the facilitation observed for two of the social contract problems. The presence of "must" led to a significant improvement in performance. The results of these experiments are strongly supportive of social contract theory in that cost-benefit information is necessary for substantial facilitation to be observed in Cosmides' problems. These findings also suggest the presence of other cues that can help guide subjects to a deontic social contract interpretation when the social contract nature of the problem is not clear. PMID- 8243032 TI - Lysis-centrifugation blood cultures in the detection of tissue-proven invasive candidiasis. Disseminated versus single-organ infection. AB - Several studies have demonstrated significantly higher frequency and more rapid detection of candidemia with blood culture methods performed by lysis centrifugation (LC) in comparison with other techniques. Little is known, however, about the ability of LC blood culture methods to detect tissue-proven invasive candidiasis. We therefore investigated the sensitivity of LC blood cultures in the detection of tissue-proven invasive candidiasis. Between 1985 and 1991, invasive candidiasis was detected in 41 (5.1%) of 803 autopsies at the Clinical Center of the National Institutes of Health (Bethesda, MD, USA). Cases were classified as single-organ (SO) candidiasis (n = 20) and as disseminated candidiasis (DI) (n = 21). Patients with DI were more likely than those with SO to have a hematologic malignancy (71% vs 15%, P < 0.001) and to have gastrointestinal mucosal candidiasis (76% vs 25%, P = 0.003). LC detected fungemia in 16 (43%) of all 37 cases with blood cultures. When analyzed by classification, Candida spp. were isolated from blood in 11 (58%) of 19 patients with DI and in five (28%) of 18 patients with SO (P = 0.13). When analyzed by number of organs infected, blood cultures were positive in seven (78%) of nine patients with > 3 organs infected by Candida in comparison to five (28%) of 18 patients with one organ infected (P = 0.024). The mean recovery time for Candida in blood cultures was 2.6 days in DI and 3.2 days in SO (P = 0.017). There was no difference in colonies of organisms per LC tube between patients with DI and those with SO.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243033 TI - Improved detection of mycobacterial antigens in clinical specimens by combined enzyme-linked immunosorbent assay. AB - Three types of antibodies against cellular and secretory-excretory protein antigens were simultaneously used for the direct detection of mycobacterial antigens in sputum and cerebrospinal fluid (CSF) specimens, using enzyme-linked immunosorbent assay (ELISA). The antibodies consisted of in-house raised and prepared anti-whole-cell, heat-killed, and sonicated Mycobacterium tuberculosis, anti-secretory-excretory protein extract of bacilli Calmette-Guerin (BCG) strain, and commercially available anti-BCG. Sputum specimens comprised 24 smear positive, culture positive, and 47 smear-negative, culture positive (SNCP), from patients with pulmonary tuberculosis, as well as 45 smear-negative, culture negative (SNCN) control samples. The CSF specimens included 18 SNCPs from patients with tuberculous meningitis and 18 SNCN controls. The sensitivity of the individual tests for sputum and CSF specimens ranged from 70% to 79% and 72% to 89%, respectively, whereas in the combined tests it reached 86%-96% for sputum specimens and 100% for CSF specimens. The specificity of ELISAs for sputum specimens was lower in the combined (73%-87%) than in the individual (87%-98%) tests, whereas for CSF specimens it was 100% in all tests. Thus, the combined ELISA approach for mycobacterial antigen detection provides a rapid and reliable laboratory adjunct in the diagnosis of patients with tuberculosis. PMID- 8243034 TI - Serum inhibitory and bactericidal activity against methicillin-resistant Staphylococcus aureus in volunteers receiving novobiocin and rifampin alone and in combination. AB - The combination of novobiocin and rifampin is effective in eliminating colonization due to methicillin-resistant Staphylococcus aureus (MRSA) and in treating experimental MRSA soft tissue infections. To evaluate novobiocin, rifampin, and the combination of the two agents for potential oral therapy in patients with MRSA infections, we measured the serum inhibitory and bactericidal activity from four volunteers against 20 MRSA strains obtained from seven different institutions. When Stratton-Reller methods employing 50% human serum were used to perform the assay, rifampin produced peak mean serum inhibitory titers of 1:40, whereas novobiocin alone produced essentially no inhibitory activity. The combination of novobiocin plus rifampin had similar inhibitory activity as rifampin alone. The bactericidal titers produced by the three regimens were significantly less than inhibitory titers. In additional studies, involving serum from five volunteers tested against seven representative strains, peak mean serum inhibitory activity of novobiocin was 1:232 when Mueller-Hinton broth was used as the diluent compared with < 1:2 when 50% human serum was used. We conclude that despite the high degree of activity of novobiocin in broth, its activity against MRSA in serum is minimal, probably related to the high degree of protein binding of that antibiotic. PMID- 8243035 TI - In vitro activity of cefpodoxime compared with other oral cephalosporins tested against 5556 recent clinical isolates from five medical centers. AB - A multicenter study was conducted in which the in vitro activity of cefpodoxime (the active metabolite of the prodrug ester cefpodoxime proxetil) was compared with those of cefixime, cefuroxime, cefaclor, cefadroxil, and clarithromycin against 5556 recent clinical isolates. Cefpodoxime demonstrated potent activity against members of the Enterobacteriaceae, in particular against species generally resistant to the established oral cephalosporins such as Proteus vulgaris [minimum inhibitory concentration (MIC)50, 0.12 microgram/ml], Providencia rettgeri (MIC50, 0.015 microgram/ml), and Serratia marcescens (MIC50, 2 micrograms/ml). Cefpodoxime was very effective against the fastidious organisms most frequently associated with respiratory infections, such as Streptococcus pneumoniae (MIC90, 0.12 microgram/ml), Haemophilus influenzae (MIC90, 0.12 microgram/ml), and Moraxella catarrhalis (MIC90, 1 microgram/ml). In contrast to other orally administrated third-generation cephalosporins (cefixime or ceftibuten), cefpodoxime demonstrated reasonable activity against oxacillin susceptible staphylococci, with MIC50 ranging from 1 to 2 micrograms/ml. All cephalosporins tested demonstrated poor activity against Pseudomonas spp., Xanthomonas spp., Enterococcus spp., and oxacillin-resistant staphylococci. Cefpodoxime had the widest spectrum of activity of all tested oral cephalosporins. PMID- 8243036 TI - In vitro activity of sparfloxacin and clinafloxacin against multidrug-resistant enterococci. AB - We evaluated the in vitro susceptibility of 140 clinical enterococcal isolates to the quinolones sparfloxacin and clinafloxacin. Isolates included Enterococcus faecalis (107), Enterococcus faecium (29), Enterococcus raffinosus (3), and one Enterococcus gallinarum. There were 111 isolates that showed high-level [minimum inhibitory concentrations (MICs) > or = 2000 micrograms/ml] resistance to gentamicin and were resistant to high levels of all other aminoglycosides; five isolates produced beta-lactamase; 21 isolates were resistant (MIC > or = 16 micrograms/ml) to ampicillin and were not beta-lactamase producers; and 13 strains were resistant (MIC > or = 32 micrograms/ml) to vancomycin. Most strains were susceptible to low concentrations of sparfloxacin and clinafloxacin, with MIC90S of 0.6 microgram/ml and 0.5 micrograms/ml, respectively. There were no inoculum effects. Time-kill experiments were performed with 22 strains; using 2 x MIC at 24 h, a > or = 2 log10 reduction in growth was observed with sparfloxacin and clinafloxacin for 14 and 17 strains, respectively. Time-kill synergism experiments were performed with 15 strains lacking high-level aminoglycoside resistance. In vitro bacterial synergism with the combination of sparfloxacin or clinafloxacin with streptomycin or gentamicin was observed for five and 12 isolates, respectively. The bactericidal activity of sparfloxacin and clinafloxacin suggest that these antibiotics may prove useful for therapy of multidrug resistant enterococci. PMID- 8243037 TI - The North American component (the United States and Canada) of an International Comparative MIC trial monitoring ofloxacin resistance. AB - Common lots of reference MIC (minimum inhibitory concentration) method reagents were used to monitor ofloxacin, a newer fluoroquinolone, and 13 other drugs against 3200 recent clinical isolates in February-April 1992. Five medical centers in the United States and Canada contributed 640 strains per facility as follows: Escherichia coli, Staphylococcus aureus, coagulase-negative staphylococci, Klebsiella spp., and Pseudomonas aeruginosa (100 strains each); Streptococcus pneumoniae (40 strains); and Enterobacter cloacae, Serratia marcescens, Salmonella spp., Haemophilus influenzae, and Moraxella catarrhalis (20 strains each). Quality-control strains were processed concurrently, MICs recorded, and data processed at a common location. Selected ofloxacin-resistant isolates were retested at a reference laboratory to confirm resistances and determine cross-resistant patterns. Results indicate the following (a) fluoroquinolones were superior in usable spectrum of activity to other orally administered drugs (for example, cefaclor, cefixime, ampicillin, amoxicillin clavulanate, minocycline, oxacillin, and trimethoprim-sulfamethoxazole); (b) ofloxacin and ciprofloxacin were generally equal to gentamicin and cefotaxime against commonly isolated Gram-negative pathogens; (c) fluoroquinolone resistance was rare among enteric bacilli, pneumococci (ciprofloxacin > ofloxacin), H. influenzae, and M. catarrhalis, but more common among oxacillin-resistant staphylococci and P. aeruginosa; (d) cross resistance was generally observed between ofloxacin and ciprofloxacin but was species or genus dependent; and (e) a new fluoroquinolone, levofloxacin, demonstrated promising activity against contemporary pathogens. PMID- 8243038 TI - Central venous catheter infection caused by Moraxella osloensis in a patient receiving home parenteral nutrition. AB - We report the first case of a central venous catheter infection caused by Moraxella osloensis, which was successfully treated without catheter removal. The isolation, identification, and pathogenesis of this species are discussed. It is recommended that Moraxella isolates be identified to species in order to determine the relative pathogenic and opportunistic roles of the various Moraxella species. Our case also demonstrates that catheter sepsis caused by some Gram-negative organisms may be amenable to systemic antibiotic therapy without the necessity of catheter removal. PMID- 8243039 TI - Bilateral septic arthritis due to Salmonella enteritidis. AB - We present an unusual case of septic arthritis due to Salmonella enteritidis involving both knee joints of a 29-year-old man with Hodgkin's lymphoma. The role of ciprofloxacin in septic arthritis caused by Gram-negative bacilli is also reviewed. PMID- 8243040 TI - In vitro activity of ampicillin, amoxicillin, ampicillin-sulbactam, and amoxicillin-clavulanic acid against consecutive clinical isolates of Enterobacteriaceae. AB - Ampicillin was generally twice as active as amoxicillin against 2440 consecutive isolates of Enterobacteriaceae from five medical centers. When beta-lactamase inhibitors were added to the penicillins, there was a significant increase in susceptibility. The magnitude of the increased susceptibility to ampicillin sulbactam (A-S) and amoxicillin-clavulanic (A-C) acid varied with the species and types of beta-lactamases elaborated. Although cross-susceptibility and cross resistance between ampicillin and amoxicillin was nearly complete, major differences were documented between A-S and A-C with 6.7% of our consecutive isolates of Enterobacteriaceae. The clinical significance of these findings remains uncertain, but they may help explain some of the discrepancies occasionally observed by clinical microbiologists with the combination drugs. PMID- 8243041 TI - In vitro activity of new quinoxaline compounds against Campylobacter species and Clostridium difficile. AB - SC-44914 and SC-44942-A are two new quinoxaline compounds with a spectrum of activity similar to that of metronidazole. We studied the activity of SC-44914 and SC-44942-A against 35 Campylobacter jejuni, 30 C. coli, and 20 Clostridium (Cl.) difficile and compared it with that of metronidazole by utilizing an agar dilution method. The quinoxalines had little activity against the C. jejuni and C. coli [minimum inhibitory concentration (MIC)90 > or = 64 micrograms/ml]. SC 44914 and SC-44942-A had excellent activity against Cl. difficile (MIC90 < or = 0.06 micrograms/ml for SC 44914, and 0.5 micrograms/ml for SC-44942A). PMID- 8243042 TI - Comparison of fetal bovine serum and colostrum-free bovine serum supplements of virus culture. AB - Incidence of virus isolation and length of time to cytopathic effect were compared in human diploid fibroblasts by using culture media supplemented with either fetal bovine serum or a less expensive alternative, colostrum-free bovine serum. The two systems were equivalent for diagnosis of 386 viral isolates. PMID- 8243043 TI - Comparison of direct specimen testing utilizing TestPack strep A with testing of specimens following a two-hour broth enrichment. AB - In a previous study, we demonstrated that a 2-h enrichment of throat swabs in broth followed by a direct fluorescent antibody test was significantly more sensitive than the Abbott TestPack Strep A for the detection of group-A streptococci. In this study, in an effort to simplify the technique, we utilized broth enrichment with an enzyme immunoassay detection method. A total of 1017 pharyngeal specimens were tested with TestPack Strep A (DTP), culture on a selective streptococcal agar, and a 2-h broth enrichment in Todd-Hewitt broth (THB) followed by TestPack Strep A (ETP). Subculture of the THB was used to arbitrate discordant test results. The sensitivities and specificities respectively were as follows: DTP (69% and 99%), ETP (90% and 98%), and SSA (97% and 100%). This enrichment method offers significantly greater sensitivity than direct testing for antigen yet still allows the potential for finalized same-day reporting of results. PMID- 8243044 TI - Gesture-speech mismatch and mechanisms of learning: what the hands reveal about a child's state of mind. AB - Previous work has shown that, when asked to explain a concept they are acquiring, children often convey one procedure in speech and a different procedure in gesture. Such children, whom we label "discordant," have been shown to be in a transitional state in the sense that they are particularly receptive to instruction--indeed more receptive to instruction than "concordant" children, who convey the same procedure in speech and gesture. This study asks whether the discordant state is transitional, not only in the sense that it predicts receptivity to instruction, but also in the sense that it is both preceded and followed by a concordant state. To address this question, children were asked to solve and explain a series of problems instantiating the concept of mathematical equivalence. The relationship between gesture and speech in each explanation was monitored over the series. We found that the majority of children who learned to correctly solve equivalence problems did so by adhering to the hypothesized path: They first produced a single, incorrect procedure. They then entered a discordant state in which they produced different procedures--one in speech and another in gesture. Finally, they again produced a single procedure, but this time a correct one. These data support the notion that the transitional state is characterized by the concurrent activation of more than one procedure, and provide further evidence that gesture can be a powerful source of insight into the processes involved in cognitive development. PMID- 8243045 TI - The roles of similarity in transfer: separating retrievability from inferential soundness. AB - Similarity is universally acknowledged to be central in transfer, but recent research suggests that its role is complex. The present research attempts to isolate and compare the determinants of similarity-based access to memory and the determinants of the subjective soundness and similarity of a match. We predicted, based on structure-mapping theory, that subjective soundness would depend on the degree of shared relational structure, particularly higher-order structure such as causal bindings. In contrast, we predicted that memory retrieval would be highly sensitive to surface similarities such as common object attributes. To assess retrievability, in three studies, subjects were asked to read a large set of stories and were later given a set of probe stories that resembled the original stories in systematically different ways; e.g., purely relational analogies, surface-similarity matches, or overall (literal similarity) matches. Subjects were told to write out any of the original stories that came to mind. To assess subjective soundness, independent subjects (and also the same reminding subjects) were asked to rate the inferential soundness of each pair; i.e., how well inferences true of one story would apply to the other. As predicted, subjective soundness was highly related to the degree of common relational structure, while retrievability was chiefly related to the degree of surface similarity. Ratings of the similarity of the pairs did not predict the retrievability ordering, arguing against the possibility that the retrieval ordering simply reflected overall similarity. Further, a fourth study demonstrated that subjects given a forced-choice recognition task could discriminate between possible matches on the basis of relational structure, ruling out the possibility that the poor relational retrieval resulted from forgetting or failing to encode the relational structure. We conclude that there is a dissociation between the similarity that governs access to long-term memory and that which is used in evaluating and reasoning from a present match. We describe a model, called MAC/FAC ("Many are called but few are chosen"), that uses a two-stage similarity retrieval process to model these findings. Finally, we speculate on the implications of this view for learning and transfer. PMID- 8243046 TI - Conceptualization of mental illness among Korean-American clergymen and implications for mental health service delivery. AB - This study examines the relationship between conceptualization of mental illness and referral intent among 50 Korean-American clergymen. Subjects are presented vignettes depicting depression, psychotic symptoms with religious delusions, and psychotic symptoms with persecutory delusions. The vignettes are followed by questions assessing problem conceptualization, cause attribution, and referral intent of mental health treatment. Respondents with a psychological conceptualization are significantly more willing to make a referral than are those who hold a religious conceptualization. Implications for effective utilization of the clergy as a mental health resource are discussed. PMID- 8243047 TI - Personal and family distress in homeless adolescents. AB - Previous research has indicated that homeless children exhibit high rates of behavioral and emotional problems and come from families characterised by conflict and rejection. Further, some evidence exists to show that family variables may relate to adolescent distress differently for homeless males and females. In this study, 117 homeless adolescents were compared to a sample of non homeless youths on the self reported incidence of personal and family problems. The homeless children reported the highest incidence of all behavioral and emotional problems, parental marital discord, overprotection, and the lowest levels of parental care and acceptance. Sex effects were not evident in reported levels of personal or family problems. However, substantially more variance in the adolescents level of behavioral and emotional disturbance was predictable from family measures for females than males. Overall, the results point to the importance of incorporating family distress models in the understanding and remediation of adolescent homelessness. PMID- 8243048 TI - A comparison of homeless men and women: different populations, different needs. AB - Homeless women are very different from homeless men, but few studies have reported data separately on them or compared them directly with men. This report on a study of 600 homeless men and 300 homeless women in St. Louis presents comparison data on these populations. The pivotal difference between homeless men and women was that unlike men, most women had young children in their custody. The women were also younger than men, more likely to be members of a minority group, and more often dependent on welfare. They had been homeless for a shorter period and spent less time in unsheltered locations. Compared to men, they had less frequent histories of substance abuse, incarceration, and felony conviction. Solitary women (without children with them), compared to women with children in their custody, were more likely to be white, had been homeless longer, and more often had a history of alcoholism or schizophrenia. On most variables, values for solitary women lay somewhere between those for men and for women with children. The population of homeless women is therefore heterogeneous, with at least two subgroups. These groups are likely to benefit from intervention programs that are designed to address their specific problems and needs, which are not necessarily the same as those of homeless men. PMID- 8243049 TI - Crisis stabilization services for children and adolescents: a brokerage model to reduce admissions to state psychiatric facilities. AB - There continues to be an increased interest in developing community-based services for children and adolescents as an alternative to inpatient care. However, there has been much more talk than action in creating such alternatives. This paper describes the success of a crisis stabilization program for children and adolescents in a community mental health center which historically over utilized the State psychiatric facility for youth. The Crisis Stabilization Program consists of three components: (1) a two-person crisis team, (2) a four person on-call team for after hours crises, and (3) funds which the crisis team utilized to broker for a wider array of community-based services. PMID- 8243050 TI - Counseling Army reservists and their families during Operation Desert Shield/Storm. AB - The hardships of war affect all members of society. Those most acutely affected are service members and their families. Civilian service personnel mobilized within an active reserve component experience a disruption of life style which can cause stress. Service members are trained to be effective soldiers, but very little training is provided to service members or their families on the wartime stresses they face. The psychiatric army reserve unit based in Minneapolis developed a family support program for three phases of wartime stress: (1) pre deployment, (2) deployment, and (3) reunification and sustainment. Recommendations are provided for family support in future wartime situations. PMID- 8243051 TI - "Aggressive" and "problem-focused" models of case management for the severely mentally ill. AB - In this paper, the relative merits and limitations of two models of case management (CM) are compared: "aggressive" and "problem-focused". Although aggressive CM has a well-established history of improving the community tenure of deinstitutionalized patients, individuals participating in this treatment are likely to become dependent on health care providers and hence require indeterminate assistance. Problem-focused CM teaches patients how to identify and resolve community-based predicaments thereby making them relatively more independent of the mental health system. Patients receiving problem-focused CM, however, need reasonably competent cognitive functions thereby ruling out participation of the most severely disabled individuals. An interaction of the two approaches is proposed in which aggressive and problem-focused CM is selected depending on the patient's current needs, cognitive deficit, and level of social support. PMID- 8243052 TI - Psychiatric care of adults with developmental disabilities and mental illness in the community. AB - Large numbers of people with co-occurring developmental disorders and mental illnesses now live in the community. Though not specifically trained to care for this population, community psychiatrists already have the necessary basic concepts and skills from working with adults with persistent mental illnesses. There are, however, some population-specific aspects of information collection, clinical assessment, diagnosis, treatment planning and application of treatment. This paper presents a practical approach to the community mental health care of people with developmental disabilities and mental disorders. PMID- 8243053 TI - The effect of taurine on the incorporation of thymidine by chick B cells. AB - 1. This study was conducted to determine the effect of extracellular taurine on chick B cell proliferation as measured by the incorporation of [3H] thymidine. 2. B cells were cultured in RPMI-1640 media containing bacterial lipopolysaccharide and either 0 or 100 microM taurine. 3. Thymidine incorporation was significantly increased in the presence of 100 microM extracellular taurine. 4. These results suggest a role for taurine in the chick B cell proliferation process. PMID- 8243054 TI - Comparative studies in molecular stability of immunoglobulin G from different species. AB - 1. Molecular stability of immunoglobulin G (IgG) from chicken and four mammalian species (cow, goat, pig and rabbit) was compared from immunochemical and biochemical view-points. 2. IgG from cow, goat and pig showed similar stability against denaturation caused by heat, acid and denaturant treatments, while rabbit IgG was shown to be more resistant to all of the treatments than other mammalian IgGs, suggesting that rabbit IgG has a highly stable molecular structure. 3. The heat- and acid-stability of chicken IgG measured by immunochemical methods were similar to those of cow, goat and pig IgG. Conformational stability of chicken IgG measured by fluorometric and enzymatic methods, however, was lower than the mammalian IgGs, suggesting that chicken IgG has a more flexible and unstable molecular structure. PMID- 8243055 TI - Fiber type composition and enzyme activities of muscles in two models of obese rats. AB - 1. In oxidative (soleus) and glycolytic (extensor digitorum longus) muscles of obese Zucker rats, a significant decrease in the percentage of relative area occupied by glycolytic fibers was observed. 2. The activity of citrate synthase and beta-hydroxy-acyl-CoA-dehydrogenase was significantly higher in muscles of obese than of lean Zucker rats. 3. In rats, 6 weeks after lesion of the ventromedial hypothalamus, no changes were observed. 4. This indicates that neither the proportion of oxidative fibers, nor the oxidative capacities are decreased in skeletal muscles of obese rats suggesting that insulin resistance cannot be ascribed to a higher glycolytic-oxidative fiber ratio. PMID- 8243056 TI - Production and characterization of recombinant turkey prolactin. AB - 1. Recombinant turkey prolactin (rctPRL) was produced as a fusion protein in E. coli, purified by affinity chromatography followed by cleavage with thrombin. The final yield of the released rctPRL (> 90% purity) was 1-2 mg/l of bacterial culture. 2. Recombinant tPRL co-migrated with the main immunoreactive band (25 kDa) in turkey pituitary extracts and was identical to natural tPRL except for the addition of three amino acids (Gly-Ser-Ser) resulting from the cloning strategy at the amino terminal end. 3. The bioactivity of the rctPRL was equipotent to ovine PRL in a rabbit mammary explant system and in the Nb2 lymphoma mitogenic assay. PMID- 8243057 TI - Comparative nuclear magnetic resonance studies of diffusional water permeability of red blood cells from different species. V--Rabbit (Oryctolagus cuniculus). AB - 1. The diffusional water permeability (Pd) of rabbit red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition with p-chloromercuribenzene sulfonate (PCMBS). 2. The values of Pd were around 6.3 x 10(-3) cm/sec at 15 degrees C, 7.0 x 10(-3) cm/sec at 20 degrees C, 8.0 x 10(-3) cm/sec at 25 degrees C, 9.1 x 10(-3) cm/sec at 30 degrees C and 10.7 x 10(-3) cm/sec at 37 degrees C. 3. Systematic studies on the effects of PCMBS on water diffusion indicated that the maximal inhibition was reached in 15 min at 37 degrees C with 0.5 mM PCMBS. 4. The values of maximal inhibition were around 71-74% at all temperatures. 5. The basal permeability to water was estimated as 1.6 x 10(-3) cm/sec at 15 degrees C, 2.0 x 10(-3) cm/sec at 20 degrees C, 2.4 x 10(-3) cm/sec at 25 degrees C, 2.6 x 10(-3) cm/sec at 30 degrees C, and 3.1 x 10(-3) cm/sec at 37 degrees C. 6. The activation energy of water diffusion was around 18 kJ/mol and increased to 27 kcal/mol after incubation with PCMBS in conditions of maximal inhibition of water diffusion. 7. The membrane polypeptide electrophoretic pattern of rabbit RBCs has been compared with its human counterpart. 8. The rabbit membrane contained a higher amount of spectrin (bands 1 and 2), while the band 6 (glyceraldehyde-3-phosphate dehydrogenase) was markedly less intense.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243058 TI - Developmental changes in the molecular forms of acetylcholinesterase during the life cycle of the lamprey Petromyzon marinus. AB - 1. We have determined the molecular forms of acetylcholinesterase (AChE) present in the skeletal muscle of the lamprey during the adult parasitic stage of its life cycle. AChE was found primarily in the globular G4 form, as well as in the asymmetric forms A4, A8 and A12. 2. We compare the complement of molecular forms present in skeletal muscle during the larval, parasitic, and spawning stages of the lamprey life cycle. The larval form, the ammocoete, contains elevated amounts of G1 and G2. However, the most striking change that we observed was in the proportion of asymmetric forms of AChE present: 5% in the ammocoete, 28% in the parasite and 9% in the spawner. 3. We speculate that these differences may be related to the physiological states of the lamprey during the various stages of its life cycle. PMID- 8243059 TI - Purification and immunological characterization of acid beta-galactosidase from dog liver. AB - 1. Dog liver acid beta-galactosidase was isolated in high yield and purified to homogeneity using a series of chromatographies on Con A-Sepharose, decyl-agarose, anion-exchange HPLC and gel-filtration HPLC. 2. Non-denaturing gel filtration by HPLC gave a single homogeneous peak corresponding to molecular mass of 180-190 kDa. During SDS-PAGE analysis, the single peak dissociated into a major band corresponding to molecular mass of 32 kDa with minor bands at 18 and 13 kDa. 3. Polyclonal antibodies raised against the purified enzyme immunoprecipitated beta galactosidase activity specifically from dog liver extracts and recognized a single 32 kDa band in Western blot analysis of dog tissue homogenates. This antibody did not crossreact with any protein band in tissue homogenates from other species examined except cat. 4. Western blot analysis of tissue extracts from dogs affected with GM1-gangliosidosis showed the presence of a 32 kDa band similar to that of controls. PMID- 8243060 TI - Guinea-pig neutrophil cationic peptides are encoded by at least three kinds of mRNA transcripts. AB - 1. cDNA clones encoding antimicrobial guinea-pig neutrophil cationic peptides, GNCP-1 and GNCP-2 were isolated from a bone marrow cell cDNA library. 2. Three kinds of cDNAs (GNCP-1A, GNCP-1B and GNCP-2 cDNAs) were obtained, and the analysis of these cDNAs indicated that GNCPs were produced as precursor proteins comprising 93 amino acid residues, which were composed of signal sequences (N terminal 19 residues), pro-peptide sequences (43 residues) and mature GNCP sequences (31 residues). 3. The deduced amino acid sequences showed that GNCP-1A and GNCP-1B differed by only one amino acid substitution in the pro-peptide region, and GNCP-1B and GNCP-2 differed by only one amino acid substitution in the mature peptide region. The nucleotide sequences of these cDNAs were highly homologous (99%). 4. Together these observations indicate that GNCPs are encoded by at least three kinds of mRNA transcripts which are likely derived from the similar genes. PMID- 8243061 TI - Defence mechanisms against free radical-induced damage in sheep, cattle and dog erythrocytes. AB - 1. Erythrocytes from sheep were to be found the least resistant to peroxidative stress induced in vitro as compared to those from cattle and dogs. 2. The differences found among species in alpha-tocopherol content and glutathione peroxidase activity were probably insufficient to explain the lowest resistance. 3. One of the main reasons of the lowest resistance may be found in the membrane composition and characteristics (membrane fluidity). PMID- 8243062 TI - Induction kinetics of immune antibacterial proteins in pupae of Galleria mellonella and Pieris brassicae. AB - 1. Pupae of Galleria mellonella and Pieris brassicae given an injection with live, non-pathogenic Enterobacter cloacae or abiotic foreign molecules induce an acquired immunity that corresponds with the synthesis of haemolymph proteins of antibacterial activity. 2. This humoral defensive response which persists for several days, differs quantitatively between insect species and between the inducers used, although very different foreign bodies induced the same immune proteins in both lepidopteran insects. 3. A stronger and longer lasting response was consistently noticed in pupae immunized with non-pathogenic bacterium than after sterile nutrient broth injections. 4. A demonstrably elevated activity of haemolymph lysozyme and trace activity of cecropins found in pupae of Galleria treated with saline W, a salt solution physiological to moths, disappear soon after 36 hr from injection. 5. In P. brassicae, however, sterile insect Ringer can give a varying, if present at all, immune response. 6. A mechanical injury (sterile wounding of insect body) can occasionally induce a similar but much weaker response. 7. The antibacterial activity was drastically reduced in Pieris or completely depressed in most pupae of Galleria when actinomycin D or cycloheximide was given at an early time post-immunization with E. cloacae. 8. It is concluded that the de novo synthesis of ribonucleic acid and immune proteins is required for expression of antibacterial activity in pupal haemolymphs. 9. The synthesis of an immune mRNA was completed about 7 hr after the injection of the immunizing bacteria. PMID- 8243063 TI - The mechanism of calcium-dependent activation of respiration of liver mitochondria from hibernating ground squirrels, Citellus undulatus. AB - 1. It is shown that Ca(2+)-dependent activation of respiration of liver mitochondria from hibernating ground squirrels is accompanied by mitochondrial swelling. 2. The swelling of mitochondria from hibernating ground squirrels, as well as the activation of mitochondrial respiration, is precluded by cyclosporin A, p-bromphenacylbromide and oligomycin. Carboxyatractiloside, on the contrary, under these conditions favors the swelling and the acceleration of respiration. 3. It was concluded that Ca(2+)-dependent activation of hibernating ground squirrel liver mitochondrial respiration resulted from the appearance of a non specific permeability pathway and from swelling of mitochondria. PMID- 8243064 TI - Oxidation reactions of human, opossum (Didelphis virginiana) and spot (Leiostomus xanthurus) hemoglobins: a search for a correlation with some structural functional properties. AB - 1. Relative to human HbA, opossum (Didelphis virginiana) hemoglobin was found to be more susceptible to autoxidation. While the initial rate of autoxidation of spot (Leiostomus xanthurus) hemoglobin is close to that of HbA, complete oxidation occurs in 50 hr. 2. Direct addition of hydrogen peroxide (H2O2) induced oxidation of hemoglobins in a definite order: spot Hb > HbA > opossum Hb. Excess H2O2 led to heme degradation and precipitation that occurred much faster for spot Hb than the case with other proteins. 3. Exposure of hemoglobins to a continuous flux of H2O2, generated by the glucose/glucose oxidase system, induced the formation of heterogeneous protein-associated oxidation products. 4. Differential reactivity among these hemoglobins under the same or different oxidative conditions, with respect to methemoglobin formation and stability of the ferric form, may reflect the differences in the local heme environment of these proteins. PMID- 8243065 TI - Developmental profile of the gene expression of a Bombyx homolog of mammalian sorbitol dehydrogenase during embryogenesis in non-diapause eggs. AB - 1. By using semi-quantitative PCR analysis, changes in the amount of the transcript for a Bombyx homolog of mammalian sorbitol dehydrogenase (BmSDH) were examined during embryogenesis in non-diapause eggs. 2. Occurrence of the transcript for BmSDH correlated with the two developmental phases, the growth of embryo and the formation of larval tissues. 3. In the first phase, an increase in the amount of the transcript for BmSDH resulted from embryonic cells rather than yolk cells. 4. In the second phase, the transcript was suggested to be abundant in fat-body cells of pharate larva, because it was abundant in fat-bodies of 5th instar larvae. 5. The difference in the expression of BmSDH gene between diapause and non-diapause eggs is discussed. PMID- 8243066 TI - Comparative mammal model of chronic rate overload: relationship of myocardial Ca cycling to heart, metabolic and lipoperoxidation rates. AB - 1. The cardiac cycle is generally believed to be timed by a sarcoplasmic Ca-cycle and powered by a sarcoplasmic ATP-cycle that uses fatty acids as fuel and generates toxic free radicals as a side-product. 2. This study used a comparative mammal approach to test this model and the hypothesis that these cycles were closely coupled and correlated with fatty acid oxidation and peroxidative injury. 3. Fatty acid oxidation and ATP-cycling rates correlated to log heart rate whereas Ca-cycling rate was directly coupled to heart rate in a one-to-one relationship. 4. Both Ca-pump and Ca-channel activities coordinately increased as heart rate increased across species. 5. Thus, Ca-cycling activity is constant across mammals, when normalized to heart rate. 6. Comparison of Ca-ATPase and Ca flux rates indicated that sarcoplasmic volume was inversely correlated with log heart rate. Basal lipoperoxidation of myocardium and susceptibility of SR to lipoperoxidation correlated with metabolic rate. 7. We identified that the horse is a metabolic outlier amongst mammals, with abnormally high fatty acid oxidation and ATP-synthetase activity compared to its heart rate. PMID- 8243067 TI - A massively parallel memory-based story system for psychotherapy. AB - We describe a memory-based system for psychotherapy, Dr. Bob, built to run on the data parallel processor Thinking Machines, Inc., CM-2a Connection Machine. The system retrieves, in parallel, stories of alcohol addiction and sexual abuse which can be used by psychiatrists in working with their patients as part of their work in recovering from addictive behavior and psychological trauma. The program is written in *LISP (pronounced Star LISP), a version of LISP used in programming Connection Machines. PMID- 8243068 TI - Use of chaos theory and complex systems modeling to study alcohol effects on fetal condition. AB - A systems dynamics computer model to predict birth complications for individual pregnant woman was developed from prospectively conducted data on a database of 125 pregnant women. The model is based upon nonlinear mathematics derived from the study of chaos and complex systems. The model was then tested prospectively on 27 additional pregnant women, making predictions on their level of obstetrical risk. The model was refined until it correctly predicted the outcomes of all 125 cases in the development database. Prediction was made with an accuracy of 25/27 cases for the prospective test cases. Predictions were made for fetal condition at birth, presence or absence of operative delivery, and presence or absence of uterine dysfunction. Then the model was used to explore alcohol use during pregnancy. A reasonable spread of alcohol use existed among subjects, allowing consideration of alcohol effects. Alcohol was found to have differential effects on fetal condition at birth depending upon the presence or absence of high levels of psychosocial stress and the use of other substances. In all cases, the effect of alcohol was only evident after the 10 drinks per week level was reached. For the high-stress/one other substance group, there could be an 18-fold effect on fetal condition at birth. For the low-stress/one other substance group, the effect was only 3-fold, and for the alcohol alone group, the effect was negligible. PMID- 8243069 TI - Monitoring the monitor: automated statistical tracking of a clinical event monitor. AB - At Columbia-Presbyterian Medical Center, a clinical event monitor processes a set of rules called Medical Logic Modules (MLMs), which generate messages (interpretations, warnings, suggestions) for health care providers. The automated statistical tracker (AST) monitors the operation of the clinical event monitor for the purpose of detecting malfunctions in MLMs or in the clinical event monitor itself. The AST follows the number of messages generated by each MLM each day and issues an alert to a system administrator if the current count of messages seems unusual compared to the MLM's past activity. The AST is based upon a combination of Poisson and normal distributions. The AST was implemented using Unix shell scripts and put into operation. Of two malfunctions that occurred during a prospective study of 12 MLMs over 85 days, the AST automatically detected one that might otherwise have gone undetected, and the system administrator detected the other during routine review of the AST's daily report. The AST's performance was compared to that of five human subjects, and it was found to rank third among the six total subjects. The AST generated eight false positive alerts during the study period (false-positive rate = 0.009 alerts/MLM day); seven of these were also picked by the human subjects. Subsequent experience has proven the AST to be useful and efficient for a system with 20 to 60 MLMs. PMID- 8243070 TI - Monitoring free-text data using medical language processing. AB - In this paper, we describe a software system for automated monitoring of free text data in a medical information system that we call RadTRAC (Radiology Text Report Analyzer and Classifier). RadTRAC uses a medical language processing tool and rules derived from statistical analysis of a database to process free-text chest X-ray (CXR) reports and identify reports that describe new or expanding neoplasms for the purpose of monitoring the follow-up of these patients. To evaluate the RadTRAC system, we examined a set of 470 consecutive radiology reports at the Veterans Administration Medical Center, Palo Alto, CA. We compared RadTRAC classification of CXR reports with retrospective expert classification of the reports and with clinical classification from CXR films as recorded in a logbook while the films were being read. The RadTRAC system had a sensitivity of 90% and a specificity of 82% using the logbook as the gold standard. This was similar to the performance of expert radiologists (sensitivity, 92%; specificity, 90%). We then reviewed the charts, appointment schedule, and subsequent X-ray reports of cases either in the logbook or that were identified by RadTRAC as needing follow-up. Two cases in the logbook could have potentially benefited from an automatic monitoring system to ensure follow-up. RadTRAC identified six confirmed new tumors or new metastatic lesions that were not in the logbook. Six other cases were identified by the RadTRAC system with suspicious X-ray findings that had either no follow-up or no further mention of the X-ray lesion in medical records. This suggests that a reminder system based on the RadTRAC technology would be potentially useful. PMID- 8243071 TI - A process for obtaining patient clinical information in the ambulatory setting. AB - The Wisconsin Ambulatory Review Project is the most extensive study to date with the goal of describing in detail the nature of care in the ambulatory setting. To assist in the description, a detailed, electronic clinical data base was created from a random sample of nonstandard, often hand-written, ambulatory patient medical records. This paper describes the set of processes that enabled significant clinical data to be collected, organized, and made available to researchers, including the collection of a representative sample of patient medical records, the creation of a detailed clinical vocabulary, the implementation of the vocabulary as a data abstraction tool, the collection and aggregation of distributed electronic data files, and the assessment of the efficiency of the abstraction process. The integration of all of these processes has produced a working system that has successfully been deployed as a pilot study across a seven-state area. PMID- 8243072 TI - A software for the description of workplaces. AB - Three softwares have been designed, the first for job histories, the second for job-exposure matrices, and this, the third and latest, for workplace descriptions. The first two softwares were presented in earlier articles while the current article presents the software for workplace descriptions. Workplace description has been based on the idea that a job is best described by listing its constituent tasks so as to determine associated risks better. The three softwares, the job history software (JHS), the job-exposure software (JES) and the workplace description software (WDS) are interfaced and constitute an integrated professional risk surveillance (PRS) system. PMID- 8243073 TI - Graphical knowledge presentation in a MUMPS-based decision-support system. AB - Conventional knowledge-based medical expert systems present information and elicit responses in the form of text. PHOENIX, a decision-support system designed to help non-radiologist physicians select diagnostic imaging procedures, offers a graphical user interface. The system constructs and displays algorithms, or flowcharts, from the rules in its knowledge base. Users can view the flowcharts and move through them by answering questions at branch points. The system provides detailed textual explanations of the rules and descriptions of the imaging procedures. The system is written in MUMPS, a common programming language for biomedical information systems, and is easily incorporated into clinical computer systems. PMID- 8243074 TI - Classification of drug-induced behaviors using a multi-layer feed-forward neural network. AB - Measurement of laboratory animal motor behavior is an important part of many studies of experimental manipulations of the central nervous system. Current automated data collection and analysis systems are limited in the number of behaviors that can be monitored and quantified simultaneously. This paper describes a signal analysis technique that when used to analyze the data from a modified Stoelting electronic activity monitor is capable of classifying multiple behavior categories automatically. In this technique, the output signal from the motility monitor is fixed-length segmented and feature extraction is performed, calculating the Fourier transform and power spectrum of each data segment. An error back-propagation neural network, implemented on a microcomputer, is used to perform behavior classification of the segment power spectra. The technique provides a high degree of accuracy in automatic behavior classification and should prove useful in the quantitative assessment of behavior. PMID- 8243075 TI - Computer analysis of eye movements. AB - An interactive program package for the acquisition, analysis and plotting of human eye movements is introduced. It is shown that the programs described in this paper can be used by scientists in a wide range of disciplines in spite of their different data analysis requirements. An example dealing with smooth pursuit tracking is given. PMID- 8243076 TI - Nonlinear identification of the PCO2 control system in man. AB - Two approaches to identification of the PCO2 system in man are described. The first uses a nonlinear 'black box' NARMAX identification package, while the second method uses a structured two-compartment Belville model. The data were obtained from volunteers breathing either room air or a controlled gas mixture, controlled via a pseudorandom M-sequence. Measurements were made of respiratory gas flow and PCO2 content of inspired and expired gases. The identification results indicate that a low-order dynamic model with nonlinear polynomial expansion gave the best fit to the data. In contrast, the Belville model gave best results with a two-compartment linear model, mainly because of difficulties in the optimisation routines when the Belville model was not linear. Thus, modern systemic methods of excitation and identification appear to be appropriate for modelling this respiratory subsystem of humans. PMID- 8243077 TI - Segmentation of medical images through competitive learning. AB - In image analysis applications, segmentation of gray-level images into meaningful regions is an important low-level processing step. Various approaches to segmentation investigated in the literature, in general, use either local information of gray-level values of pixels (region growing based methods, for example) or the global information (histogram thresholding based methods, for example). Application of these approaches for segmenting medical images often does not provide satisfactory results. Medical images are usually characterized by low local contrast and noisy or faded features causing unacceptable performance of local information based segmentation methods. In addition, because of a large amount of structural information found in medical images, global information based segmentation methods yield inadequate results in region extraction. We present a novel approach to image segmentation that combines local contrast as well as global gray-level distribution information. The presented method adaptively learns useful features and regions through the use of a normalized contrast function as a measure of local information and a competitive learning based method to update region segmentation incorporating global information about the gray-level distribution of the image. In this paper, we present the framework of such a self organizing feature map, and show the results on simulated as well as real medical images. PMID- 8243078 TI - A computer package for the multivariate nonparametric rank test in completely randomized experimental designs. AB - When research data are measured on at least an ordinal scale and the assumptions required in the theory underlying parametric statistical methods are in question, nonparameteric procedures based on the ranks provide a sound approach to statistical analysis. Biomedical investigations, especially clinical trials, typically involve multivariate response and therefore multivariate statistical methods are called for in the interpretation of results. We discuss applications of the nonparametric multivariate rank test for completely randomized designs. Large sample theory can be used to support these statistical methods for assessing group differences in location. In small samples, randomization tests provide a basis for inferences. The execution of the procedure is facilitated by a computer program developed by the authors. PMID- 8243079 TI - Prompt i.v. anticoagulation of cerebral embolus based on clinical information alone: a case report. PMID- 8243080 TI - Ovarian abscess as an unusual infectious complication of laser vaporization of the cervix: a case report. AB - Laser vaporization of the cervix has been reported to have an extremely low incidence of infectious morbidity compared to other treatment modalities. This case report describes the development of an ovarian abscess following laser surgery of the cervix in a patient with no known risk factors for disease. PMID- 8243081 TI - Quadriplegia--some problems and some modern solutions. PMID- 8243082 TI - New developments at the Connecticut Peer Review Organization. PMID- 8243083 TI - Oral ciprofloxacin, ofloxacin, and lomefloxacin as alternatives to intravenous antimicrobial therapy. AB - Ciprofloxacin, ofloxacin, and lomefloxacin are oral fluoroquinolones with ideal characteristics for oral antimicrobial therapy. These agents provide 1) appropriate and reliable serum concentrations, 2) broad antimicrobial activity, 3) proven efficacy in the treatment of serious infections, and 4) good tolerability. Although cations decrease the absorption of these drugs, there are proven means of avoiding these interactions. Therefore, these agents could easily be utilized as alternatives to prolonged intravenous treatment of many infectious diseases in patients who do not have conditions which impair drug absorption. PMID- 8243084 TI - Response to Dr. Arthur's Koffler's case report. PMID- 8243085 TI - Managed competition. PMID- 8243086 TI - A shortage of nurses or a surplus of people called nurses? PMID- 8243087 TI - Resistance to change as a contribution to medical progress. 1943. PMID- 8243088 TI - Dr. Johnson and his burden of illness. PMID- 8243089 TI - Relief of benign essential blepharospasm and ? memory loss by cyproheptadine. AB - These results in a small group of patients seem quite clear for the benefit of cyproheptadine in patients with BEB, especially where botulinum A injections fail, are too expensive, or are refused. The apparent memory improvement requires further study. A search for more effective serotonin antagonists without the anticholinergic and antihistaminic side effects of cyproheptadine is needed and is in process. PMID- 8243090 TI - Homogenization of syncytial tissues. AB - This article derives a continuum representation of a multicellular, syncytial tissue directly from its microstructure and basic physical principles. The final equations for the homogenized syncytium contain the bidomain model as a special case. The derivation begins with an idealized, periodic representation of the tissue microstructure. Potentials inside the cells and in the extracellular fluid are governed by the Laplace equation. Electrical properties of the membrane separating those two regions are represented by the boundary conditions on the potentials. An homogenization process based on a two-scale asymptotic expansion converts this microscopic, pointwise description into an averaged, continuum representation by two reaction-diffusion equations. The same process also yields formulas for the effective conductivities of the tissue in terms of its microstructure and specific conductivities of cytoplasm and extracellular fluid. The validity of the homogenized syncytium model is assured deep in the tissue for autonomous processes, such as propagation, and in the presence of external fields that are nearly uniform and limited in strength. The derived model is not formally valid on the surface of tissue, in the proximity of sources, and under strong or rapidly changing electrical fields. PMID- 8243091 TI - Biomechanics of the knee joint: a critical review. AB - The literature concerning kinematic and kinetic studies on the knee joint is comprehensively reviewed in this article. Also reviewed are studies of etiology and operative treatment of injury, as well as chronic disease such as dislocation, arthritis, and ligamentous rupture. This paper formulates experimental study and mathematical model analysis of the tibio-femoral and patello-femoral joints, respectively. The sections on experimental study cover the following: the tibio-femoral joint including load bearing capacity, knee laxity, ligamentous strain, articular geometry, and multiaxial movement; the patello-femoral joint including forces and stresses and patellar tracking patterns. Further, three items, considered as future problems, are discussed briefly: individual variations in material properties of the soft tissue and biphasic cartilage-bone structures, quantitative description of bone geometry, and quantitative determination of extreme tensions and distortions in connective tissues surrounding the knee. PMID- 8243092 TI - Muscle sound: bases for the introduction of a mechanomyographic signal in muscle studies. AB - Muscular sound is a mechanical phenomenon detectable at the surface of an active muscle, which has been known and described since 1800. Only recently, because of the availability of reliable transducers and sophisticated analysis techniques, has this signal become attractive for monitoring the mechanical aspects of muscle contraction. The muscular sound characteristics were investigated both during electrically elicited and voluntary contractions. In the first case, the influence of the biophysical and mechanical properties of the muscle on this signal was studied. During voluntary efforts the summation of the mechanical activity of the recruited motor units was analyzed. The results indicate that the muscular sound may be an adjunct tool to the force, the physiological force tremor, and electromyogram to obtain information on the muscle mechanical model as well as on muscle motor control. This review focuses on the following aspects of the signal: (1) recording problems; (2) muscle sound properties during stimulation of isolated and in vivo muscle; (3) time and frequency domain analysis during non fatiguing and fatiguing contractions; (4) comparison with other signals related to muscle activity; (5) discussion on the origin; and (6) possible practical applications. PMID- 8243093 TI - Power spectral analysis of heart rate variability: a noninvasive signature of cardiac autonomic function. AB - Power spectral analysis of short segments of beat-to-beat heart rate variability (PS/HRV) reveals three distinct peaks. In human PS/HRV, the high frequency (HF) band (0.15 to 0.4 Hz) is correlated with respiratory driven vagal efferent input to the sinus node. The low frequency band (LF) 0.06 to 0.15 Hz is believed to be due to baroreceptor mediated blood pressure control. Therefore, PS/HRV represents a noninvasive signature of the balance between sympathetic and parasympathetic components of the autonomic nervous system. This paper reviews the literature on the methodological issues relevant to signal processing, computational, and clinical applications of PS/HRV. Factors affecting the power in the LF and HF bands are examined in healthy controls. Recent work from several laboratories suggests that PS/HRV is a potentially powerful tool for exploring neurocardiac dysfunction in patients with a variety of cardiac and autonomic disorders. Mathematical models which simulate neurocardiac control are examined. Concerns regarding the lack of standardization between different laboratories are expressed. As the PS/HRV attains the status of a clinical diagnostic test, we hope that this review serves as a source of integrated information for researchers in this field. PMID- 8243094 TI - Surface electromyogram during voluntary contraction: processing tools and relation to physiological events. AB - Surface electromyography (SEMG) has been used extensively in the last years in a variety of applications, including muscle function assessment, pathology identification, ergonomics, pattern analysis, or population characterization. Advanced processing methods, especially in the spectral domain, provide the research worker with more and more precise and user-friendly tools for signal characterization, analysis, and classification. The use of such sophisticated tools requires many assumptions on signal characteristics, and the wide variety of computing options related to each processing method makes it difficult to compare the results of different works when these options are omitted in the reports or improperly applied. This work first aims at taking stock of the various processing methods which have emerged in the last years around surface electromyography: signal acquisition, random feature extraction, time and spectral parameter determination, statistical tests application. The main methods are briefly explained and discussed, then variations between apparently equivalent methods are pointed out, necessary hypotheses are underlined, and the use of such methods in SEMG processing is shown with respect to the more recent works. A second section shows how authors deal with parameters extracted from SEMG in order to relate them to physiological modifications (force, fiber type, fiber environment). PMID- 8243095 TI - The nickel controversy at home and abroad. PMID- 8243096 TI - A dermatological diary. Portrait of a practice. PMID- 8243097 TI - Psoriasis and ultraviolet radiation. AB - Prevention and detection screening programs as a public health service in curtailing the ever-increasing incidence of all forms of skin cancer are reviewed. The effect of solar and artificial ultraviolet radiation on the general population and persons with psoriasis is examined. PMID- 8243098 TI - Diaper dermatitis: an approach to prevention employing effective diaper care. PMID- 8243099 TI - Calcinosis cutis following electromyographic examination. AB - We report the first case of calcinosis cutis following electromyographic examination. All previously reported cases of iatrogenic calcinosis cutis occurred following electroencephalographic examination using calcium-containing electrode compounds or following the extravasation of calcium-containing solutions along infiltrated intravenous lines. We also review the literature on calcinosis cutis due to iatrogenic and occupational exposure to calcium salts. The clinical presentation of calcinosis cutis is variable, but the histologic appearance is diagnostic. This entity may be underreported because of its benign, self-limited course and lack of recognition by physicians. Calcinosis cutis should be recognized by family practitioners, dermatologists, and subspecialists who encounter patients who have undergone prolonged monitoring studies that involve the use of electrodes. PMID- 8243100 TI - Systemic antifungals. AB - The administration, effectiveness, action, adverse reactions, and commentary are presented in Table I for a selected number of systemic antifungal agents. PMID- 8243101 TI - Lyme disease: the evolution of erythema chronicum migrans into acrodermatitis chronica atrophicans. AB - Erythema chronicum migrans and acrodermatitis chronica atrophicans are both recognized to be lesions associated with Lyme disease, although they are thought to be distinct entities. In this paper, the clear evolution of erythema chronicum migrans into acrodermatitis chronica atrophicans is demonstrated. PMID- 8243102 TI - Lichen planus of the nails. AB - The case of a nineteen-year-old man with lichen planus limited to the nails is reported. He showed no skin or mucous membrane involvement. PMID- 8243103 TI - Unusual eruption of cutaneous metastases from occult gastric carcinoma shortly after HIV-1 infection. AB - We report a case of eruptive metastases to the skin of the face and neck from a hitherto unknown gastric carcinoma, occurring in a fifty-two-year-old man (a regular blood donor), about six months after he contracted human immunodeficiency virus-1 infection. The eruption, consisting of more than thirty pea-sized cutaneous nodules appeared within about twenty days. The patient complained of epigastric pain and weight loss. Histologic examination showed signet-ring cells of gastric carcinoma metastases. Gastroscopic examination showed a widely infiltrating tumor. No metastases were found elsewhere after a careful screening. Immunologic variables were only slightly altered. This case and a brief review suggest that gastric carcinomas, possibly as well as other coincidental tumors, may present atypically during human immunodeficiency virus-1 infection. PMID- 8243104 TI - Pityriasis rubra pilaris presenting as subacute cutaneous lupus erythematosus. AB - Diseases of a very different nature may occasionally present in a similar manner. Differentiating among several possible diagnoses may be complicated by confusing and conflicting laboratory data. Our recent evaluation and treatment of a patient confirmed the need for thorough testing, particularly in patients suspected of having a connective tissue disorder. PMID- 8243105 TI - Inhibitory effect of cisplatin on the enzymatic peroxidation of platelet endogenous arachidonate. AB - The effect of cisplatin at concentrations of 0.1-33 microM on the enzymatic transformation of arachidonate in pig blood platelets was investigated. Estimation of the level of malonyl dialdehyde (MDA) after stimulation of pig platelets with thrombin (endogenous arachidonate pathway) and exogenous arachidonate served as an indicator of the transformation. MDA concentration was determined by means of a modified method with thiobarbituric acid. The effect of cisplatin on the thrombin-induced formation of MDA in blood platelets was dependent on the time of platelet preincubation with the drug. It was reduced significantly (p < 0.001) after 30 min action of cisplatin on the platelets. In contrast, cisplatin had no effect (p > 0.05) on the arachidonate pathway dependent on the activity of thromboxane synthase. PMID- 8243106 TI - Axenic cultivation of a pathogenic Phytomonas species isolated from tomato fruit, and from its phytophagic insect vector, Phthia picta (Hemiptera: Coreidae). AB - Axenic cultures of Phytomonas sp. were obtained from naturally infected tomatoes and from Phthia picta, a predator of tomato plants, by using a biphasic medium with Roitman's complex medium overlaying rabbit blood-agar slants. Light and electron microscopy of both isolates showed a similarity of morphological characteristics among the flagellates in fresh material or after cultivation. Other properties, including their agglutinability with the haemolymph of Phthia picta, suggest that these isolates are virtually identical. PMID- 8243107 TI - Aflatoxin induced testicular degeneration and decreased meiotic index in Rattus norvegicus. AB - The effect of oral consumption of 200 +/- 100 ppb of crude aflatoxin (B1, B2, G1, G2) showed testicular degeneration and a decrease in the meiotic index. PMID- 8243109 TI - The effects of aging on fixed DNA. AB - Agarose-gel electrophoretic analysis was carried out to study the effect(s) of aging on fixed naked DNA as well as on the DNA part of fixed chromosomes. Results show that aging of fixed DNA produces alterations in molecular size; the alteration is more effective in naked DNA than in the DNA part of fixed chromosomes; and the alteration occurs to a greater extent in air-dried DNA than in DNA aged in fixative. Since B-mercaptoethanol is capable of preventing DNA alterations induced by air-drying, oxidative processes are invoked for explaining these findings as well as the effect(s) of aging on fixed cytological preparations. PMID- 8243108 TI - High and low affinity 5-HT2 and 5-HT1C binding sites: responses to neonatal 5,7 DHT lesions in rat brain. AB - 5-HT receptor denervation supersensitivity has been proposed to explain behavioural supersensitivity to L-5-HTP in rats with 5,7-dihydroxytryptamine (5,7 DHT) lesions. No upregulation of 5-HT2 binding sites was found despite supersensitivity to putative 5-HT2,1C drugs. To test the hypothesis that the 5 HT1C properties of these drugs are involved instead, dose-response and time course studies of 5-HT1C and 5-HT2 receptors were performed using several different radioligands in rat brain after making neonatal 5,7-DHT lesions by intraperitoneal injection. 5-HT1C sites labelled with [3H]-mesulergine showed a distinct regional distribution: brainstem > diencephalon > cortex > hippocampus > cerebellum, constituting 65, 70, 31, 70, and 73% of total sites labelled by [3H] mesulergine in the absence of 20 nM spiperone to block 5-HT2 sites, respectively. 5,7-DHT lesions did not significantly alter BMAX, KD, or nH of [3H]-mesulergine labelled 5-HT1C sites in cortex or other regions but did reduce the density of cortical [3H]-paroxetine sites (-55%). Cortical 5-HT1C sites labelled by [3H]-5 HT or [3H]-mianserin, and cortical 5-HT2 sites labelled by [3H]-DOB or [3H] ketanserin, were also unaffected. These data suggest that although denervation supersensitivity of 5-HT1C or 5-HT2 receptors may occur at the level of the receptor transducer-effector, there is no evidence it occurs at the receptor recognition site. PMID- 8243110 TI - Expression of protein kinase C isoforms in HL60 and phorbol ester resistant HL525 cells. AB - Isoform protein expression and activity were investigated to evaluate whether differential expression of protein kinase C (PKC) accounts for the inability of HL525 cells to respond to phorbol 12-13,myristate acetate (PMA). Immunoblotting analysis of lysates from both the parental HL60 and PMA-resistant HL525 cells revealed expression of alpha-, beta-, delta- and zeta-PKC isoforms. Diminished expression of delta-PKC was observed along with increased expression of zeta-PKC in HL525 cells. Lysates from HL60 cells were resolved into the following 4 peaks of PKC activity using hydroxylapatite: peak 1, beta; peak 2, delta; peak 3, alpha; and peak 4, zeta. A similar elution pattern was noted for PKC isoforms expressed in HL525 cells although no peak 2-delta activity was recovered. PMA treatment of HL60 cells abolished activity of peaks 2, 3, 4; diminished alpha-PKC and increased zeta-PKC expression. In contrast, PMA treatment of HL525 cells did not abolish activity of peaks 1, 3 or 4, compared with the parental HL60 cells and failed to stimulate changes in zeta-PKC isoform expression. These data suggest that the inability of PMA to induce differentiation of HL525 cells may reside in diminished expression of delta-PKC along with alterations in zeta-PKC. The findings support the hypothesis that differential expression of PKC isoforms in HL525 cells contributes to the lack of morphological changes in response to PMA. The data also demonstrate that PMA alters the level of histone-dependent phosphotransferase activity as well as PKC cofactor dependence. PMID- 8243111 TI - Acute morphological effects of cocaine on rat cardiomyocytes. AB - The cardiovascular actions of cocaine in vivo are varied and often antagonistic. Cocaine produces excitatory sympathomimetic effects by interfering with re-uptake of norepinephrine at adrenergic nerve terminals. However, the local anaesthetic properties of cocaine exert depressant effects on the myocardium. In an attempt to differentiate the direct effects of cocaine from the indirect sympathomimetic effects on the myocardium, primary cultures of neonatal rat cardiomyocytes were established under serum-free conditions and exposed to cocaine and/or norepinephrine. After 36-48 h in culture, spontaneously contracting cells were treated with cocaine (10-1,000 micrograms/ml) and contractile rate (beats/min) quantitated, after which the cells were processed for ultrastructural examination. The contractile rate was reduced at all dosages with nearly 80% reduction at the highest concentration studied. Recovery of beating rate was observed 24 h after removal of cocaine. Pronounced cytoplasmic vacuolation of the cells occurred at concentrations > or = 100 micrograms/ml. Ultrastructural examination revealed extensive myofibrillar disruption, membrane damage, and a near complete loss of organized sarcomeres. Nuclear morphology remained unaffected. Within 24 h after removal of cocaine from the medium, myocytes recovered their characteristic cytoplasmic architecture, indicative of sarcomere reassembly. The results observed in response to cocaine were distinctly different from the response to norepinephrine. In these myocytes, the contractile rate was enhanced twofold and morphological damage was not observed. These findings suggest that cocaine can directly alter myocardial morphology independent of its sympathomimetic effects. PMID- 8243112 TI - Regulation of the phosphorylation of histones and glycogen synthase. AB - The phosphorylation of histones and glycogen synthase by protein kinases was analysed by SDS-polyacrylamide gel electrophoresis and autoradiography. The phosphorylation of histone III-S by the catalytic subunit of cyclic AMP-dependent protein kinase (A-PK) or cGMP-dependent protein kinase (G-PK) was inhibited by archidonic acid, sphingosine and staurosporine. Using the catalytic subunit of A PK, the phosphorylation of histone VIII-S was inhibited by Ca2+, arachidonic acid and staurosporine; the phosphorylation of histone II-S was inhibited by phosphatidyl ethanolamine, phosphatidyl inositol, arachidonic acid and staurosporine; and the phosphorylation of glycogen synthase was inhibited by arachidonic acid and staurosporine. After being phosphorylated by the catalytic subunit of A-PK, calpain II with 4 microM Ca2+ was less effective in degrading histone III-S, which had been prephosphorylated by PK-C. PMID- 8243113 TI - Retinoic acid treatment inhibits mitosis in the pre-existing spinal cord during tail regeneration of the axolotl larva, Ambystoma mexicanum. AB - During tail regeneration in the axolotl larva, Ambystoma mexicanum, retinoic acid reduced mitosis some 10-fold in the pre-existing spinal cord. This conclusion followed from comparisons of mitotic indices in the central grey matter of spinal cords, proximal to the amputation plane, of animals intracoelomically injected with (a) retinoic acid 2 days before amputation and the mitotic inhibitor, demecolcine 5 h before sacrifice; or (b) as for the latter but without retinoic acid; or (c) only the solvent for (a) and (b). Animals were sacrificed concurrently, 9 days after tail amputation. In addition, the demecolcine treatment revealed the true mitotic activity of the spinal cord of axolotl to be at least five times greater than might otherwise have been suspected. PMID- 8243114 TI - The impact of alcohol and other drugs on the child welfare system. AB - The Child Welfare League of America surveyed its member agencies on the impact of alcohol and other drugs (AOD) on the delivery of child welfare services. It discovered that the impact is profound and adversely affects the system by compounding problems, such as personnel shortages and shortages in the availability of foster homes, which existed before AOD was identified as a major social problem. Research is needed that more accurately describes the number and characteristics of children and families who enter the child welfare system, that contributes to improved screening and assessment, and that explicates the systemic changes and stresses that result from the impact of alcohol and other drug abuse. PMID- 8243115 TI - Perpetrator gender and type of child maltreatment: overcoming limited conceptualizations and obtaining representative samples. AB - A lack of sensitive differentiation across the types of child maltreatment and lack of attention to differences due to perpetrator gender are serious shortcomings in child maltreatment perpetration research. Findings about perpetration may be confounded and of questionable validity as a result. Certain conceptual and methodological difficulties lie at the root of this lack of sensitivity and lead to inadequate research designs and sampling procedures. These difficulties are discussed in this article, with corrective recommendations and an example of a new approach that will enhance sensitivity to the effects of both perpetrator gender and types of maltreatment. PMID- 8243116 TI - Present status of renal transplantation in China. PMID- 8243117 TI - Advances of homotransplantation of testes in China. PMID- 8243118 TI - Prevalence of human papillomaviruses 16 and 18 in transitional cell carcinoma of bladder. AB - It has been speculated that human papillomaviruses (HPV) might be a causative agent of transitional cell carcinoma of the bladder. With the polymerase chain reaction technic, fresh specimens of 53 transitional cell carcinomas of the bladder were examined, and bladder epithelium from 12 patients with hyperplasia of prostate and peripheral blood lymphocytes of 8 normal individuals served as controls. The primers set for HPV-16 and 18 were selected from E6/E7 regions of open reading frames. HPV-16 was positive in 52.8% of the specimens examined, whereas HPV-18 positive in only 3.7%. No controls showed either HPV-16 or 18. Reports on the role of HPVs in carcinogenesis of bladder are scanty and yet there exist conspicuous discrepancies among them. There is a diversity of opinions on the contamination and variation of technical procedures. More work should be done before the relationship between HPV-16 and development of bladder carcinoma can be clarified. PMID- 8243119 TI - Treatment of chronic allograft rejection. AB - Chronic renal allograft rejection at present the most important factor affecting long-term graft survival, and effective therapeutic method is still lacking. This article reports 43 cases of chronic renal rejection treated from January 1989 to April 1992. Effective result was obtained in 27 cases (62.8%). The authors believe that increasing dosage of the immunosuppressants is an effective method for the management of chronic renal rejection and the earlier the treatment is given the better the results would be. The selection and the regulation of the dosage of the immunosuppressants should be determined according to the individual situation. PMID- 8243120 TI - Long-term results of surgical treatment for acute hemorrhagic necrotizing pancreatitis. AB - Eighteen patients with acute hemorrhagic necrotizing pancreatitis who survived from operation were followed up for 12 to 43 months. Late complications including pancreatic external fistula, pseudocyst, hyperglycemia, etc were reviewed. The pancreatic juice was collected through endoscopic cannulation. The volume and HCO3- concentration of the pancreatic juice and three kinds of pancreatic enzymes were measured. The results demonstrated that inspite of anatomic abnormality or functional impairment of the pancreas, the pancreatic insufficiency rarely manifested to be marked because of the compensative ability of the viable exocrine pancreas. PMID- 8243121 TI - Different methods for wound management of radiation-burn injury in rats. AB - Different methods of burn wound management were used in rats with severe radiation-burn injury. The 30-day survival rate and the functional recovery of thymocytes and splenocytes were significantly higher in the group with early escharectomy and skin grafting than in the group without operational intervention and in the group with skin grafting in the recovery stage. In the group with early escharectomy and skin grafting on the 1st day after injury, all the autografts survived and the survival of the homografts was also markedly prolonged. Escharectomy and skin grafting performed in the late stage aggravated the wound condition, and infection and hemorrhage were the main factors hastening the death of the skin grafts as well as the animals. The findings indicate that escharectomy and skin grafting in the early stage after injury are conducive to the recovery of immune function and consequently to the recovery of the whole organism because infections are minimized after escharectomy and closure of the wound surfaces by skin grafting. PMID- 8243122 TI - CT detection of wrist bone erosion in rheumatoid arthritis. AB - A correlative radiographic and CT study of the wrists of 30 patients with rheumatoid arthritis (RA) was reported. The results showed that sensitivity of CT in detecting wrist erosion is superior to that of conventional radiograph. Accordingly, wrist CT scanning is useful for early diagnosis of RA. PMID- 8243123 TI - Reperfusion arrhythmias in acute myocardial infarction. AB - In 27 patients with acute myocardial infarction (MI) and their infarct-related coronary artery being completely occluded who received thrombolytic therapy or percutaneous transluminal coronary angioplasty, reperfusion was confirmed by immediate coronary angiography in 24. Reperfusion arrhythmias (RA) occurred in 19 (79.2%) of the patients, including ventricular arrhythmias in 13 (54.2%). Ventricular fibrillation and sustained ventricular tachycardia developed in 2 (8.4%) and accelerated idioventricular rhythm in 5 (20.8%), the latter was a reliable indicator of coronary recanalization. Transient sinus bradycardia or AV block occurred in 10 (66.7%) of the 15 patients with inferoposterior MI. The occurrence of RA was not related to the duration of ischemia; ventricular RA was also not related to the location of MI and the occurrence and severity of ischemic arrhythmias before reperfusion. PMID- 8243124 TI - Molecular cloning, sequencing and expression of core and NS3 fragments of HCV from patients with HCV infection. AB - Fragments of core and NS3 of hepatitis C virus-Hunan (HCV-Hun) were cloned by RT PCR and gene recombinant techniques from blood samples collected in Hunan Province, China. In comparison with sequences of our samples with those of HCV-US and HCV-J, the homologies of nucleotides and amino acids were about 90%, indicating that fragments of core and NS3 of HCV-Hun were in a relative conserved region of HCV. Two fusion proteins containing the peptides coded by HCV core (MBP HCV core) and HCV. NS3 (MBP-HCV. NS3-Gal) were expressed by Escherichia Coli with recombinant plasmids. The specific HCV antigenicity of the two fusion proteins were identified by western blotting. Therefore, MBP-HCV. core and MBP-HCV.NS3-Gal were found useful for anti-HCV assay. PMID- 8243125 TI - Purification and characterization of human hepatic stimulator substance. AB - Human hepatic stimulator substance (hHSS) was purified from fetal liver with 6,000-fold decrease in protein content and 840-fold increase in specific growth stimulative activity. Purification procedures included the heating of a homogenate in 35% (W/V) Tris-HCL at 95 degrees C for 20 min, high and ultra speed centrifugation, passage over Sephadex G100 gel filtration, DEAE-cellulose ion exchange, TSK G3000 SWG high performance liquid chromatography (HPLC) and YWG C 18 reverse phase HPLC techniques. The most purified material (HP-HSS) revealed cell-specific and dose dependent increase in 3H-TdR incorporation into cellular DNA. As little as 38 ng of the HP-HSS per ml of culture medium produced a 2.5 fold increase in DNA synthesis. Further studies indicate that HP-HSS in combination with insulin and epidermal growth factor stimulate DNA synthesis 16 fold compared with serum and hormone free controls and nearly 3-fold over hepatoma growth with HP-HSS alone. Sodium dedecyl sulfate polyacrylamide gel electrophoresis with silver stain and ultrascan XL laser densitrometer quantitative scanning revealed only one band at 12,800. PMID- 8243126 TI - A comparative study on the solid phase capture--platelet test, platelet adhesion immunofluorescence test and lymphocytotoxic test. AB - The sensitivity, specificity and reproducibility of the solid phase capture platelet test (CPT) were compared with those of the platelet adhesion immunofluorescence test (PAIFT) and the lymphocytotoxic test (LCT). This study shows that CPT has a higher sensitivity than PAIFT (P < 0.001) and LCT (P < 0.01). The specificity was 86% lower than LCT (100%) (P < 0.01). Both the CPT test and PAIFT were comparable with regard to reproducibility (100%, 92%). The solid phase assay is easier to handle than PAIFT and LCT, allowing identification of HLA as well as of platelet specific antibodies. It is an efficient antibody screening test. PMID- 8243127 TI - Endolaser photocoagulation after vitrectomy on rabbit's eyes. AB - Ophthalmoscopic and histopathologic studies of endolaser photocoagulation after vitrectomy were carried out on rabbit's eyes. The endoprobe was connected to the Coherent "Novus 2,000" argon laser system. Under appropriate application the grayish white retinal lesions produced by argon green and argon all wavelength endolaser photocoagulation were similar ophthalmoscopically at a treatment power of 80-100 mw and exposure duration of 0.2s. Histopathologically, the characteristic changes of the retinal lesions produced by argon all wave-length were more significant than that produced by argon green endolaser photocoagulation. Scattered retinal photocoagulation can be done when the opaque medium is cleared on the operating table. PMID- 8243128 TI - Candida zeylanoides causing candidiasis as tinea cruris. AB - Candida zeylanoides caused candidiasis as tinea cruris in 4 cases. Candida zeylanoides were characterized by good growth of the strain in malt medium, potato dextrose agar and Sabourard's agar. Their colony was milky white and greasy with smooth surface. The diameter of the colony reaches 12 mm within a week. As candida type pseudohyphae but not hyphae, they do not produce ascospore and bolospore. Carbon sources fermentation showed positive results. Nitrogen sources assimilation agar (KNO3) negative, arbutin agar negative, nonvitamin medium positive. G+C mol% value of their DNA was 51.24. Animal experiment showed that guinea pigs were infected by Candida zeylanoides. The main feature of candidiasis as tinea cruris caused by the strain was pink infiltrative erythema with clear margin, a lot of grain-sized papules, a few vesicles, and thin scales. The disease can be cured by preparation of ketoconazole. PMID- 8243129 TI - Phagocytosis of platelet in coronary heart disease patients with blood-stasis syndrome. AB - Phagocytosis of platelets was studied under electronmicroscope in 36 healthy persons and 64 coronary heart disease (CHD) patients with blood-stasis syndrome (BSS). The result showed that both the number of latex particles phagocytized by platelet and platelets which phagocytized latex particles were decreased in CHD with BSS. It was also found in this group that most of the platelets having active-phagocytosis were of medium size. The proportion of large platelets containing plentiful glycogen granules was much more in CHD patients with BSS and the shapes of platelets were changed readily with active exocytosis and aggregations. Therefore the present findings of disability of platelet phagocytosis might be considered the basic change in CHD with BSS. PMID- 8243130 TI - Genetic identification for routine diagnosis of HBV by dot-blot hybridization. PMID- 8243131 TI - Coloanal anastomosis in radical surgery for low rectal carcinoma by intra-anal circular ligation. AB - Using self-made special supporting tube, coloanal anastomosis was performed by intra-anal ligation for radical surgery in 48 patients with low rectal carcinoma. The anastomoses were completed without suturing, and the results were quite satisfactory. Follow-up after operation revealed that the bowel movement and the sphincteric function were excellent in all patients. The anal canal and the anal sphincter were all kept intact and abdominal colostomy was not required for decompression. Anastomosis without suturing, least trauma, and short operation time made blood transfusion unnecessary during the operation. This technique is actually derived from Park's operation, and is considered a promising procedure for coloanal anastomosis in the radical surgery for low rectal carcinoma. PMID- 8243132 TI - Microsurgery in orthopaedics. PMID- 8243133 TI - Recurrent lumbar disc herniation: what recurs? (A morphological study of recurrent disc herniation). AB - A morphological study was conducted on disc material obtained during surgical discectomy in 20 patients. The series was divided into 4 groups. Group I: 3 patients who had never been operated on before; group II: 3 patients who had already been submitted to discectomy but at a level which differed from that of the disc examined; group III: 4 patients treated surgically for contralateral herniation; group IV: 10 patients with true recurrent disc herniation, that is, already submitted to surgery for the treatment of homolateral herniation of the same disc. All of the cases submitted to surgery had been operated on more than a year previously. Moreover, the disc material obtained during autopsy in a patient operated 15 years before for disc herniation was studied. A morphological study of the cases and the data reported in the literature reveals that the fibrocartilaginous tissue of recurrences differs from that of primary herniation because of the presence of large collagen bundles associated with fibrillar framework. This aspect suggests the hypothesis that is also supported by the autoptic material examined that recurrent disc herniation is formed by repair tissue that substitutes the part of the disc that is operated on, and that recurrent herniation is caused by mechanical collapse of the repair tissue. From a surgical point of view, a distinction may be made in terms of early recurrences, which appear within one year of surgery and are formed by disc tissue which is identical to that of primary herniation, and late recurrences, the subject of this morphological study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243134 TI - Considerations on seven cases of breakage of the polyethylene acetabular cup. AB - The authors describe seven cases of breakage of the polyethylene acetabular cup observed in 6 patients (5 females and 1 male) after a period of time ranging from 2 to 11 years postsurgery. By comparing the results obtained with those reported in the literature the authors analyze the possible etiopathogenesis of this rare late complication in total hip arthroplasty, which they particularly believe to be caused by hyperwear and fatigue phenomena of the acetabular cup, in turn caused by mechanical overloading. PMID- 8243135 TI - Contracted valgus flat foot caused by tarsal synostosis in the adolescent. AB - Contracted valgus flat foot in the adolescent is frequently caused by tarsal synostosis or synchondrosis which are often not revealed in common radiographic views, and which may instead by clearly shown by CT scan. These synostoses are prevalently symptomatic during adolescence, when by ossifying they block the subtalar joint in valgus; they are less symptomatic between 20 and 40 years of age. Pain returns during the fifth decade due to the occurrence of secondary arthrosis of the nearby joints. PMID- 8243136 TI - Spondylocostal dysplasia (Jarcho-Levin syndrome). AB - The authors report 3 cases of rare spondylocostal dysplasia (Jarcho-Levin syndrome) associated with multiple visceral anomalies (V.A.C.TE.R.L. syndrome). The difficulty encountered in treating the disease because of the complexity and multiplicity of the visceral anomalies, and the extent and type of associated vertebral deformities is emphasized. The cases described revealed a moderate amount of progression, and were able to be treated non-surgically. It is also emphasized that even when there is moderate scoliosis associated costal deformities may cause anti-cosmetic thoracic gibbosity. PMID- 8243137 TI - Agenesia of the pectoral muscle. AB - Two cases of agenesia of the pectoral muscle are presented. Clinical examination is sufficient for diagnosis. Nonetheless, dosage of LDH and CPK is needed in order to exclude primary myopathy. Surgery is rarely indicated in selected cases of female sex, for cosmetic purposes. PMID- 8243138 TI - Association between fracture of the diaphysis of the femur and lesion of the ligaments of the knee. AB - The authors evaluated 97 knees out of a series of 135 fractures of the femur in 130 patients, revealing 34 cases of capsuloligamentous laxity, equal to 35% of the cases. No specific correlation between type of trauma, type and site of fracture, and lesion of the ligaments was observed. The frequency of unrecognized lesions, particularly in the posterior capsule, was made evident. The frequency of ligamentous lesions rises to 44.4% when fracture of the femur is associated with fracture of the homolateral tibia as compared to 26.7% in cases of isolated fracture of the femur. Recurvatum, which was present in a fair number of the cases, was related more to post-fracture therapy and rehabilitation rather than to specific capsuloligamentous lesion of the knee. PMID- 8243139 TI - Lowe syndrome: general problems in a female patient. AB - The authors describe a clinical case of a patient affected with Lowe syndrome. Ocular modifications, as well as those involving the nervous system, renal function, and the locomotor apparatus, typical of the syndrome, are taken into consideration. The problem of axial deformity of the knee, secondary to spontaneous fracture of the femur, a frequent occurrence in these patients, is dealt with. PMID- 8243140 TI - Orthopaedic manifestations of leukemia during childhood: a rare case of isolated periosteal reaction. AB - The authors describe one case of acute lymphoid leukemia during childhood manifested by migrant arthralgias. The only radiographic signs were constituted by a periosteal reaction in the left fibula. The main clinical symptoms and radiographic signs described in the literature and used to diagnose leukemia are reported. PMID- 8243141 TI - Quiz. Osteoblastic osteosarcoma of high grade malignancy. PMID- 8243142 TI - [Reducing the occurrence and development of pregnancy induced hypertension, lowering maternal and infant mortality rate]. PMID- 8243143 TI - [Analysis on the causes of maternal death with pregnancy induced hypertension in Shanghai from 1981 to 1990]. AB - The total deliveries in Shanghai from 1981 to 1990 were 1,770,103 and there were 456 cases of maternal death. Among them, 42 cases were caused by pregnancy induced hypertension (PIH) which accounted for 9.21% and ranked fourth in the causes of maternal death in Shanghai. The maternal mortality of PIH was 2. 37/10(5). The cerebrovascular accident and heart failure were the leading causes which accounted for 66.67%. The study showed that strengthening prenatal care, supervising 3 main symptoms and signs of PIH, paying attention to mean arterial pressure (MAP), using antispasm drugs such as MgSO4 in proper way and timely termination of pregnancy are the key points of decreasing maternal death of PIH. The average MAP of 18 cases died of cerebrovascular accident was 17 kPa (1kPa = 7.5 mmHg). Among them, MAP > or = 18.7 kPa (140 mmHg) was found only in 4 cases which accounted for 22.22%. It suggested that the threshold value of cerebrovascular accident at the level of MAP > or = 18.7 kPa may be too high and need to be further studied. PMID- 8243144 TI - [The use of synthetic haANP III in the treatment of heart failure in pregnancy induced hypertension]. AB - Highly active atrial natriuretic peptide III (haANP III) was administered for the treatment of heart failure due to pregnancy induced hypertension (PIH) in 7 patients with success. The heart failure was rapidly controlled within 24-48 hours with lowering of the blood pressure, disappearance of edema and urinary protein and alleviation of subjective symptoms. The plasma level of renin, angiotensin, aldosterone (RAA) and SOD all decreased. The results suggested that haANP III had the ability to facilitate the excretion of sodium and water, dilate the blood vessels and inhibit the action of RAA, and it could effectively reduce heart load and improve the cardiac function. Therefore, haANP III seemed to be an ideal new drug in treating heart failure in PIH, and it would have a wide scope for future development. PMID- 8243145 TI - [Cerebral lesions in the severe pregnancy induced hypertension: 61 cases of X-ray computed tomography of the brain]. AB - Among 61 patients with severe pregnancy induced hypertension, cerebral lesions were detected on CT scans in 23 cases (37.7%). The positive rates were 5/25 cases in preeclampsia and 18/36 cases in eclampsia. The incidence rate of cerebral lesions in eclampsia was significantly higher than that in preeclampsia (chi 2 test, P < 0.05). Furthermore, it showed that patients with renal impairment and retinal changes were more susceptible to cerebral lesions (P < 0.05), through comparative study of the relationships between either the function of liver and kidney or retinal changes and cerebral lesions. Main manifestations of cerebral lesions were ischemia, edema and infarction. The represented the different pathological stage of cerebral lesions. The cerebral lesions were mainly involved at cortical or subcortical area of bilateral parietal or occipital lobe (60%), secondly at the deep basal ganglia and the superior sagittal sinus. The pathological process of cerebral lesions and the management of pregnancy induced hypertension were analysed. PMID- 8243146 TI - [The relation between the levels of serum lipid peroxide, superoxide dismutase and atrial natriuretic peptide in placenta, umbilical cord vein and intrauterine growth retardation in pregnancy induced hypertension]. AB - This paper was written on the following base: 31 women with severe pregnancy induced hypertension (PIH) were chosen randomly as an observation group, 32 healthy and gestational age matched pregnant women with uneventful delivery were served as control group. Blood samples were collected from intervillous space of placenta and umbilical cord vein after delivery. The levels of LPO, SOD and atrial natriuretic peptide (ANP) were measured separately. From the placenta point of view, the pathogenic mechanism of PIH and the relation to the occurrence of intrauterine growth retardation (IUGR) were discussed. The results showed: the concentrations of LPO, SOD and ANP of placenta and umbilical cord vein were higher in the PIH group than that in the control group. The difference between 2 groups was statistically significant. A positive correlation between LPO and ANP was found both in PIH group and in control group (r = 0.641, P < 0.02). The birth weight of newborn was 2,174 +/- 142 g in PIH group and 3,460 +/- 187 g in control group. The difference between them was significant (P < 0.01). It suggested that the pathogenesis of PIH and the occurrence of IUGR might be closely related to the high concentration of LPO in placenta and umbilical cord vein. PMID- 8243147 TI - [Acute renal failure in severe pregnancy induced hypertension: a report of 19 cases]. AB - Twenty-six pregnant women complicated with acute renal failure (ARF) were admitted to Beijing Friendship Hospital during Jan. 1972 to Dec. 1990, the incidence rate was 0.045%. Among them, 4 cases of ARF were due to non-specific factors to pregnancy (15.4%) and 22 cases (84.6%) were due to factors correlated with pregnancy. Severe pregnancy induced hypertension (PIH) was the main cause of ARF in late pregnancy accounting for 86.4% (19 cases). In this series, 7 cases with eclampsia and 12 cases with preeclampsia, the incidence rate of ARF in preeclampsia and eclampsia was 0.91% and 11.3% respectively. The average age was 28.3 yr. 9 out of 19 cases were parous women. During pregnancy and labour, patients had more than one complications. 4 cases each were complicated with abruptio placentae, postpartum hemorrhage, intracranial hemorrhage and serious puerperal infection respectively. 3 cases were complicated with HELLP syndrome and one each with acute fatty liver, hypertension or gentamycin nephrotoxicity. 10 cases had deliveries preceded admission. The cesarean rate was 52.6% (10 cases). ARF onset before labour in 9 cases and postpartum in 10 cases. Developed only in 1 case of postpartum ARF in our department, this patient had a twin gestation, also complicated with severe PIH and acute fatty liver. Auria and oliguria ARF were found in 18 cases, nonauria in only 1 case. In this study, the highest serum BUN was 7.5-39.3 mmol/L (21-110 mg/dl). Medical management and rectal dialysis or peritoneal dialysis were performed in 10 cases who were in early stage, mild ARF or those ARF occurred before admission.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243148 TI - [Characterization of two antigens OC41 D and OC41 E associated with ovarian carcinoma]. AB - Two antigens, OC41D and OC41E, were characterized by two monoclonal antibodies (COC41D and COC41E) generated with the antigens (OC41) as immunogen, which were dissociated from immune complex in a patient with ovarian carcinoma. The expressing rates of the antigen OC41D and OC41E were 70.2% and 52.5% in epithelial ovarian cancer. They were mainly detected on the membrane of the cancer cells, also a little in the plasma. Two antigens were found no cross reactivities with HIA, CEA, alpha-FP, and beta-MG. Their express were not related to cell circle. Antigen OC41D might be a protein with molecular weight of 62,000 and could not be denatured by heating. Antigen OC41E might be a glycoprotein with molecular weight of 15,000. These two antigens may be new antigens highly associated with ovarian carcinoma. PMID- 8243149 TI - [Nucleolar organizer region-associated proteins and estrogen receptor of endometrial adenocarcinoma]. AB - The nucleolar organizer region-associated proteins (AgNOR) staining technique and estrogen receptor (ER) detective technique were applied to endometrial adenocarcinoma in grade 1-4 and endometrial cystic hyperplasia. The results showed that there was no significant difference of AgNOR numbers between the grade I endometrial adenocarcinoma and endometrial cystic hyperplasia, while a significant difference of the number, size and localization of the nuclei was observed between endometrial adenocarcinoma and grade 2-4 endometrial adenocarcinoma (P < 0.001). The ER content in well differentiated endometrial adenocarcinoma was more than poor differentiated ones. PMID- 8243150 TI - [Pathological analysis of adenomatoid tumours in the female genital tract]. AB - A review of 18 cases of adenomatoid tumours in the female genital tract was carried out. There are 14 cases in uterus, 3 in fallopian tubes and 1 in ovary. The incidence of adenomatoid tumour in uterus, fallopian tubes and ovary is 3.1%, 0.5% and 0.2% respectively. The result showed that the uterus adenomatoid tumour is more common than fallopian tube and ovary (P < 0.01). All of the 18 cases are found in internal genital tract. The incidence of adenomatoid tumors is increased with the size of surfaces serosa or germinal epithelium in different part. Majority adenomatoid tumour are closing to the folding region of coelomic epithelium or midline of uterus. These findings suggest that adenomatoid tumour may be originated from mesothelia. PMID- 8243151 TI - [Actinomyces-like organisms infection in intrauterine devices wearers]. AB - The prevalence of actinomyces-like organisms in cervicovaginal smears of 2,327 Chinese women, including 1,279 intrauterine devices (IUD) wearers and 1,048 non wearers, was investigated. Cervical smears were stained by the Papanicolaou method for the microscopic examination of actinomyces-like organisms. The relationship between actinomyces-like organisms infection and occupations, age, duration of IUD wearing and clinical symptoms were analysis. The result showed that the overall detection rate of actinomyces-like organisms in 2,327 women was 0.69%. The detection rates in IUD wearers and non-wearers were 1.1% and 0.2%, respectively, which were significantly different (P < 0.01). The rate of actinomyces-like organisms infection was significantly higher in women wearing IUD for more than seven years. Whereas the occupation or age of women did not affect the detection rate significantly. PMID- 8243152 TI - [Changes in the endothelial system and the level of endothelins in women with pregnancy-induced hypertension]. PMID- 8243153 TI - [Radioimmunoassay of plasma endothelin in maternal and fetal umbilical cord vein in pregnancy induced hypertension]. AB - Endothelin (ET) level was measured in 48 cases of normal pregnancies and in 29 cases of pregnancy induced hypertension (PIH) by radioimmunoassay (RIA). The results indicate that the difference of ET values between maternal and fetal umbilical cord vein was nonsignificant in normal pregnancy. Though a slight increase in maternal ET level was found in mild and moderate PIH, however the difference was nonsignificant when compared with normal pregnancy. ET increased significantly in plasma of patients with severe PIH. ET levels of fetal umbilical cord vein in different groups showed no much change. It suggests that ET increasing in plasma of patients with PIH may play a role in pathogenesis of PIH. PMID- 8243154 TI - [Primary report of a specific light absorption peak of red blood cells in pregnancy induced hypertension--one drop blood assay with photoacoustic spectroscopy]. AB - One drop of red blood cells (RBC) was assayed with double beam photoacoustic spectroscopy device (PAS) in 89 blood samples collected from 71 pregnant women during 3rd trimester of pregnancy. A specific light absorption peak at a spectral range of 620-650 nm was showed in 86.36% of women with pregnancy induced hypertension (PIH), while only 16.67% of normal pregnant women (P < 0.01). However we can not identify the component which induced this specific peak in RBC. Our conclusion is that this simple assay could be used promisingly in prediction and differential diagnosis for PIH in clinic. PMID- 8243155 TI - [Effect of 3,4-dihydroxyacetophenone treatment on intrauterine growth retardation]. AB - In this paper, the results of a pilot study of 3,4-dihydroxyacetophenone (DHAP) treatment for intrauterine growth retardation (IUGR) were reported. 20 out of 38 cases of IUGR were treated with DHAP and 18 with amino acid. Additionally, 170 normal pregnant women were served as control group. The following parameters have been observed and measured including uterine fundal height (UFH), body weight (BW), S/D ratio of umbilical artery (UmA), hemorheological indices, platelets aggregation, TXB2/6-keto-PGF1 alpha ratio and also fetal and neonatal various growth indices etc. After administration of DHAP, all the parameters almost restored to the normal range. The results expressed that the therapeutic effect of DHAP was much better than that of amino acid. It has also been verified by neonatal birth weight and fetal biparietal diameter. The clinical effective rate of DHAP treatment group was 90.00% which was significantly higher than that 74.00%, 79.00% of amino-acid treatment group. Meanwhile, the mechanism of DHAP has preliminarily been discussed. PMID- 8243156 TI - [Correlation between serotonin and prostaglandins in peritoneal fluid collected from patients with pelvic congestion]. AB - The concentration of serotonin and prostaglandins in peritoneal fluid were measured in 14 patients with pelvic congestion and 14 normal women. Results showed that in pelvic congestion group 5-HT and TXB2 and 6-keto-PGF1 alpha levels were higher than that of control; but DA and histamine were lower. It is suggested that the serotonin and prostaglandins may play an important role in developing pelvic congestion. PMID- 8243157 TI - [The clinical and pathological characters of malignant neoplasms arising from endometriosis]. AB - Eight cases of malignant tumors arising from endometriosis were reported; the ovary was the primary site in 7, whereas extragonadal site (rectum) in 1. The histologic classification was as follows: endometrioid carcinoma in 1. Clear cell carcinoma in 2, mixed clear cell carcinoma and endometrioid carcinoma in 2, mixed papillary cystadenocarcinoma in 1 and serous tumor of borderline malignancy in 1. The histologic pattern arising from endometriosis of the rectal wall was endometrioid adenocarcinoma with benign endometriosis contiguous to it. It was found that the ectopic endometrial tissue may undergo malignant change when they were under stimulation by some factors. One of which may be intrinsic or extrinsic estrogen, which may be related to the high incidence rate in this age group. Laparotomy should be done for assessing malignant change when ovarian mass was greater than 10 cm or grew rapidly. Five year survival rate will be expected to improved with postoperative progestin or irradiation. Preventive measures were also suggested. PMID- 8243158 TI - [Effect of postoperative chemotherapy on the reproductive function and offspring health in patients with ovarian neoplasms]. AB - The reproductive function and their offspring health in 30 patients with ovarian neoplasms following preservation of tubo-ovarian and uterine functions operation, and adjuvant or preventive chemotherapy were studied. All the patients were followed up for 2-18.5 years, 65.5% of the patients showed normal cycle of menstruation. The patients under 25 years old, they were demonstrated a significant higher frequency of abnormal menstruation than that over 25. In this series 15 patients wish to have child, 12 of them were pregnant (a gestational rate of 80%), totally there are 20 gestations, including 13 fullterm deliveries, 1 premature delivery, and 7 artificial abortions, however, no spontaneous abortion occurred. Fourteen healthy offsprings were followed up to date, 11 of them are normal in health, the Gesell tests of intelligence were also normal, as well as the G-band karyotype analysis. Only the SCE frequencies of 3 objects in one family comprised the mother and her two daughters were significant higher than that of the control. We recommend that women who preserving reproductive function desired to be pregnant at least 2 years later following chemotherapy, SCE determination must be studied before any attempt to become pregnancy. PMID- 8243159 TI - [Stage Ia cervical cancer]. AB - One hundred and fifteen patients with stage I cervical cancer were operated and studied retrospectively. By comparing the original standard of stage Ia (stromal invasion less than 3mm) and the new FIGO definition and analysing the relationship among stage of the cancer, lymphatic metastasis and prognosis, no lymphatic metastasis was found in 25 cases of original stage Ia, and no recurrence during the five year's follow up. There were five cases with lymphatic, metastasis in 42 cases of new FIGO definition and 3 cases recurred and died during the five year's follow up. It is pointed out that the new FIGO definition of stage Ia fails to provide guidelines for choice of treatment and can not differentiate the stage I a1 and stage I a2. If 3mm stromal invasion can be used as a criteria to differentiate stage I a1 and I a2, then we may have a clear and acceptable standard which may be helpful for the choice of treatment. PMID- 8243160 TI - [The phytohemagglutinins and soybean agglutinin in diagnosis of endometrial adenocarcinoma]. AB - The distribution of PHA and SBA receptors was studied by ABC immunoperoxidase method in 30 cases of endometrial adenocarcinoma (EA), 10 cases of normal endometrium (NE) and 20 cases of cystic glandular hyperplastic endometrium (CGH). The results showed that the positive rate of PHA and SBA in EA was 46.67% and 73.33% respectively, but negative and low in NE and CGH (P < 0.05). PHA and SBA showed a progressive rising tendency along with the changes of NE to CGH and EA (P < 0.01). It is suggested that both PHA and SBA may be used as differentiated markers in EA and may be helpful in clinical diagnosis. PMID- 8243161 TI - [Ripe cervix and cervical ripening]. PMID- 8243162 TI - [Advances in the research on group B streptococcal infections in obstetrics]. PMID- 8243163 TI - Organization of centromeres in the decondensed nuclei of mature human sperm. AB - The localization of centromeres in mature human sperm was shown by immunofluorescent labeling and nonisotopic in situ hybridization. In the decondensed nucleus structural elements (dimers, tetramers, linear arrays and V shape structures) formed by individual centromeres of nonhomologous chromosomes were observed. They organize the compact chromocenter, which was shown for nuclei decondensed to a low extent. The chromocenter is buried inside the nucleus; in contrast, telomeric regions of chromosomes were tentatively localized on the periphery. Thus, a gross architecture, which can influence selective unpackaging of the paternal genome upon fertilization, exists in human sperm. PMID- 8243164 TI - Amplification of the nucleolus organizer region during the sexual phase of Neurospora crassa. AB - Previously we have shown that the nucleolus organizer region (NOR) of Neurospora crassa displays frequent size changes during crosses. In these initial studies, we observed that decreases in NOR size are far more common than increases. Here, we have investigated the inheritance of NOR size in a strain with an unusually small NOR. We call this strain SNO for small nucleolus organizer. We found that progeny that inherit their rDNA from SNO receive either an NOR that is larger than that of SNO or, rarely, the same size, but never an NOR that is smaller than that of SNO. The number of progeny that inherit their NOR from SNO is not significantly different from the number that inherit their NOR from the other parent in the cross. This argues against the idea that the failure to find progeny with NORs smaller than that of SNO is due to inviability of spores carrying such an NOR, or that it is due to unconscious bias by the experimenter against isolating such spores. These results can most easily be explained by a combination of unequal sister chromatid exchanges in the rDNA, or sister chromatid conversion, coupled with selection against nuclei harboring small NORs during the premeiotic phase of the Neurospora life cycle. Other, less conventional, explanations are also possible, such as "directed" increase in the target NOR without corresponding loss at some other NOR. PMID- 8243165 TI - Partial reconstruction of the lampbrush loop pair Nooses on the Y chromosome of Drosophila hydei. AB - We present the analysis of genomic DNA fragments that were isolated as potential segments of the lampbrush loop pair Nooses on the short arm of the Y chromosome of Drosophila hydei. More than 300 kb of DNA were recovered in BamHI lambda and cosmid clone groups. This DNA is composed of the Y-specific ay1 family of repetitive DNA sequences, and of other repetitive DNA sequences, which at least in part are also located elsewhere in the genome (Y-associated sequences). Two additional classes of DNA fragments were obtained from an EcoRI library. One of them consists of ay1 repeats without apparent interspersion, including a total of more than 300 kb of DNA. The other is composed of tandemly repeated YsI sequences, a Y-specific sequence derived from ay1. This class includes more than 400 kb of DNA, which is also not interspersed by other sequences. Our results show that only the ay1 repeats interspersed by Y-associated DNA sequences can represent parts of the 260 kb transcription unit forming the lampbrush loop, whereas the ay1 and YsI repeats without interspersion form separate and nontranscribed clusters of repetitive DNA. PMID- 8243166 TI - Localization of the lampbrush loop pair Nooses on the Y chromosome of Drosophila hydei by fluorescence in situ hybridization. AB - We have used fluorescence in situ hybridization to map the positions of the different repetitive DNA sequences from the region forming the lampbrush loop pair Nooses on the Y chromosome of Drosophila hydei. This region harbours a megabase cluster of tandemly organized repeats of the Y-specific ay1 family and a megabase cluster of tandem repeats of the related Y-specific YsI family. In addition, ay1 repeats also occur in short blocks that are interspersed by other repetitive DNA sequences that we call Y-associated, since they have additional copies on other chromosomes. Using specific probes for ay1, YsI and Y-associated DNA sequences, we show that there is one large proximal cluster of YsI repeats and one, more distally located, large cluster of ay1 repeats. The Y-chromosomal copies of the Y-associated sequences are located in the most distal part of the ay1 cluster. This is consistent with the juxtaposition of ay1 and Y-associated sequences in more than 300 kb of cloned genomic DNA. Since both ay1 and Y associated sequences have been shown to be transcribed in the Nooses, the lampbrush loop is formed in a distal region of the short arm of the Y chromosome, adjacent to the terminally located nucleolus organizer region. The clusters of homogeneous ay1 and YsI repeats are of no functional significance for the formation of the lampbrush loop. PMID- 8243167 TI - The sex determining region of Chironomus thummi is associated with highly repetitive DNA and transposable elements. AB - The dominant male sex determiner in chromosome III of the midge Chironomus thummi thummi is closely linked to a large cluster of tandem-repetitive DNA elements, the Cla elements, which are otherwise highly repetitive and distributed over more than 200 sites on all chromosomes. Chromosome III displays a hemizygous cluster of Cla elements in males but not in females. The chromosomal location of this hemizygous Cla element cluster is in the region of the male determiner M as localized by cytogenetic analysis. With Cla elements as hybridization probe, it was possible to clone a large part of the sex determining region. Molecular analysis of the DNA of males and females in this region displayed a number of differences between the two sexes. One striking difference is an unusual transposable element associated with the male sex determining region. The sex determining region also contains several other tandem-repetitive DNA elements in addition to the Cla elements. They are interspersed with single copy DNA. The accumulation of repetitive elements in the sex determining region interpreted as the result of a lack of recombination between the male/female heteromorphic region, although recombination in the other sections of chromosome III occurs. PMID- 8243168 TI - Two new X-autosome Robertsonian translocations in the mouse. II. Sex chromosome configurations in spermatocytes of hemizygous males. AB - The influence of X-autosome Robertsonian (Rb) translocation hemizygosity on meiotic chromosome behaviour was investigated in male mice. Two male fertile translocations [Rb(X.2)2Ad and Rb(X.9)6H] and a male sterile translocation [Rb(X.12)7H] were used. In males of all three Rb translocation types, the acrocentric homologue of the autosome involved in the rearrangement regularly failed at pachytene to pair completely with its partner in the Rb metacentric. The centric end of the acrocentric autosome was found regularly to associate either with the proximal end of the Y chromosome or with the ends of nonhomologous autosomal bivalents; the proportions of cells with such configurations varied between pachytene substages and genotypes. Various other categories of synaptic anomaly, such as nonhomologous synapsis, foldback pairing and interlocks, affected the sex chromosome multivalent in a substantial proportion of cells. In one of the Rb(X.12)7H males screened, an unusual, highly aneuploid spermatocyte that contained trivalent and bivalent configurations was found. Rb translocation hemizygosity did not appear to increase to a significant extent the incidence of X-Y pairing failure at pachytene, although the incidence was elevated at metaphase I in Rb(X.12)7H animals. Overall, a comparison of the frequencies and types of chromosome pairing anomalies did not suggest that these were important factors in the aetiology of infertility in males carrying the Rb(X.12)7H translocation. PMID- 8243169 TI - Cytogenetic and molecular aspects of position-effect variegation in Drosophila melanogaster. V. Heterochromatin-associated protein HP1 appears in euchromatic chromosomal regions that are inactivated as a result of position-effect variegation. AB - Position-effect variegation in Drosophila melanogaster is accompanied by compaction of the corresponding chromosomal regions. The compaction can be continuous, so that bands and interbands located distal to the eu-heterochromatic junction fuse into one dense block, or discontinuous, when two or more zones of compaction are separated by morphologically and functionally normal regions. In this work it was found that in both continuous and discontinuous compaction the blocks of dense material contain the immunochemically detectable protein HP1, which has previously been characterized as specific for heterochromatin. The regions undergoing compaction do not contain HP1 when they have a normal banding pattern. Thus, it may be proposed that HP1 is one of the factors involved in compaction. If two different or two identical rearrangements are combined in the same nucleus, they variegate independently. The frequency of compaction of the two rearrangements in the same nucleus corresponds to the product of the frequencies of the compact state of the individual elements. The extent of compaction (i.e. the number of bands involved in heterochromatization) of each rearrangement does not depend on the compaction pattern of the other rearranged element. PMID- 8243170 TI - [The experiment of tumor-inhibiting effect of green tea extract in animal and human body]. AB - Green tea extract (GTE) inhibits obviously gastrointestinal malignant tumors in rats induced by N-methyl-N'-nitro-n-nitrosoguanidine (MNNG). The incidence of tumor was 93.3% in the MNNG positive control group, whereas 53.3% in the MNNG plus GTE group (P < 0.01). The percentage of inhibiting tumor growth was 88% (P < 0.001). N-nitrosoproline blockage test in 3 groups of 45 cases of patients from the highly prone to (gastric cancer people), examined with gastroscope and classified pathologically, revealed that GTE can block N-nitroso-proline synthesis in vivo, with blockage rate of 91% in superficial chronic gastritis (P < 0.001), 94% in intestinal metaplasia (P < 0.001) and 93% in atypical hyperplasia and gastric cancer (P < 0.001). Chemical analysis demonstrated that the main ingredients in GTE are catechols (with the content of 50%), vitamins and trace elements. The mechanism of GTE's cancer-inhibiting and cancer-preventing function might be that catechols combine with N-nitroso-compounds or resolve them, and reduce their carcinogenic activity. These results will be of great significance in preventing gastrointestinal cancer. PMID- 8243171 TI - [Study of anti-lipid peroxidation of vitamin E in human body]. AB - Anti-lipid peroxidization in vivo has been observed by VE per os. The results have proved that contents of serum VE and RBC VE obviously were increased and that contents of serum LPO and RBC LPO obviously were decreased in normal adults and patients with acute hepatitis taking VE (200 mg/d) after 10 days. Specially, there was significant change in the patients with acute hepatitis group (53 cases). Increased levels of VE in serum and RBC were 137% and 97% respectively. Decreased levels of LPO in serum and RBC were 48% and 20% respectively. The study showed that VE has very strong anti-lipid peroxidization. PMID- 8243172 TI - [Inhibitory effects of garlicin and cinnamaldehyde on SOS response and their mechanisms in Escherichia coli]. AB - Inhibitory effects of garlicin and cinnamaldehyde on SOS response and their mechanisms in Escherichia coli were investigated. Garlicin and cinnamaldehyde suppressed the lambda cI depended SOS response induced by 4-nitroquinoline-N oxide (4NQO), mytomycin (MMC) and methyl methane-sulfonate (MMS), and the lexA depended SOS response induced by 4NQO and UV to various extent, respectively. They also diminished the rexA 441-depended SOS response induced by temperature (at 42 degrees C) However, they did not show any effect on the constitution of SOS functions. It suggested that this inhibitory effect could be via RecA protease which regulates the cleavage of lexA repressor, and perhaps, lambda cI repressor. PMID- 8243173 TI - [Report of food poisoning by Salmonella hadar in China]. AB - A food poisoning epidemic caused by Salmonella hadar was confirmed by epidemiologic investigation and laboratory examination Samonalle hadar had been detected in the specimens of food feces of patients. Many epidemics of food poisoning caused by Salmonall group of have been reported before, but that caused by the serogroup Hadar hereby reported organisms is the first of its hind in China. Attention should be paid by the public health authorities. PMID- 8243174 TI - [The genotoxicity of air particles tested by SOS chromotest]. AB - Samples of variously-sized total suspended particles in the air of one sampling site of TaiYuan city were collected. The samples were extracted with acid and a simulated lung fluid (SLF) respectively. Both extracts mainly consisted of metallic elements. Using SOS chromotest as a means to test the gentoxity of the extracts and five metallic compounds, namely Cr6+, Ni2+, Pb2+, Mn2+ and Cd2+ were tested first. All the compounds tested could induce the SOS response to various extent, showing that the method was sensitive to metallic compounds. For the extracts of air particles, both the extracts of acid and of SLF of the smaller sized particles could induce SOS response. This indicated the existence of metaklic genotoxicants in the smaller-sized particles. Being convenient to use fast and precise, the SOS Chromotest has its unique advantage for detecting carcinogenic metallic compounds. PMID- 8243175 TI - [Study on distribution of the Pseadomonas cocovenenans subsp farinofermentans Meng ZH&Wang DS et al in the natural environment of Hebei Province]. AB - In this paper, the distribution of the Pseudomonas Cocovenenans Subsp Farinofermentans Meng ZH&Wang DS et al in the natural environment of Hebei Province is reported. Several food poisoning cases caused by the bacterium have taken place, but it is still unusual in the natural environment such as soid and food. Fresh tremella and mushroom are heavily polluted by the bacteria. New knowledge is gained in the differential diagnosis between the poisoning caused by poisonous mushroom and this bacteria. Control measures are suggested for the prevention and cure of the food poisoning caused by this bacteria. PMID- 8243176 TI - [Investigation of chronic arsenic poisoning caused by high arsenic coal pollution]. AB - This article reports the results of an investigation on environmental arsenic pollution and chronic arsenic poisoning in a rural area. Exploitation of high arsenic coal caused drinking and irrigating water to be polluted by arsenic and burning of this coal caused severe environmental arsenic pollution including air, food, soil and drinking well water. 1548 villagers in 47 villages suffered from chronic arsenic poisoning who used this coal in daily life. The polluted air and food were mainly responsible, while the polluted drinking water and skin absorption played some part in poisoning. When arsenic level in coal is as high as 100mg/kg, we should consider the possibility of environmental arsenic pollution and chronic arsenic poisoning in exposed population. The high arsenic coal's distribution is very uneven. When controlling the disease, it is important to remember monitoring the quantity of arsenic coal outside the arsenic coal mining area. PMID- 8243177 TI - [An immunological study of workers exposed to diphenylmethane diisocyanate (MDI)]. AB - The average (OD values) and positive rates of specific IgE antibodies to a MDI human serum albumin (HSA) conjugate in 25 workers exposed to MDI were significantly lower than those in 8 subjects of MDI asthma, while they were significantly higher than those in the 30 control subjects who had never bee exposed to MDI before. The positive rates of specific IgE antibodies in symptomatic MDI-workers were significantly higher than those in asymptomatic MDI workers. The results of studies of specific IgE antigenic inhibition of serum ELISA activity confirmed the antigenic specificity to MDI-HSA. The average levels and positive rates of serum total IgE in 25 MDI-workers significantly increased when compared with 30 non-exposed subjects; while no significant difference when compared with 8 subjects of MDI asthma. Agreement rates of specific IgE were 85.7% with skin-prick test. The percentage of the acid alpha-naphthyl acetate esterase activity in T-cell of peripheral blood showed no statistically significant difference in all 3 groups. The results of the study suggested that the level of specific IgE was related to exposure to MDI and IgE mediated response played an important role in the allergic mechanism due to MDI, while it was not related to cell-immunological response. PMID- 8243178 TI - [A preliminary enquiry into the normal values of vision in motor vehicle drivers]. AB - A study on distant vision, visual sense and colour vision in 2,218 motor vehicle drivers in Inner Mongolia Region was carried out and the effect of age on vision was analysed. The results showed that vision in drivers diminish with increasing age and the age threshold for visual acuity was 45 years. This paper discusses the normal values of vision in two age groups of pre-and-after the threshold age line, and these results may offer a scientific basis for drawing up occupational examination standards for drivers at their posts or under training. PMID- 8243179 TI - [Investigation of iodine intake in Tianjin resident]. AB - 83 kinds of main food on the market in Tianjin were determined using alklin ashing method. The iodine intake of adult in Tianjin was found to be 113.76 mg/day which is below the recommended daily intake. It is not adequate for pregnant and lactational women. Its deficiency may possibly affect the intellectual development of infants. Hence sufficient attention should be paid by public health authorities. PMID- 8243180 TI - [Changes in epidemic features of epidemic cerebrospinal meningitis after vaccination with purified meningococcal polysaccharide vaccine group A in Zhengzhou]. AB - The authors studied the epidemic features of epidemic cerebrospinal meningitis before and after mass vaccination with Purified meningococcal polysaccharide vaccine group A. After the mass vaccination, the morbidity rate of epidemic cerebrospinal meningitis fell considerably year after year. The epidemic cycle changed for the first time. The ratio of the number of cases in the city to those in the countryside was reduced. The phenomenon that the infectious disease spread along traffic lines from city to countryside disappeared. The cases mainly occurred in the remote mountain village. The morbidity rate in the group of 0-3 years relatively increased. The level of antibody titers in population obviously increased. The carrier rate of meningococci and constituent ratio of group A reduced. The epidemic group was group A, but cases of group C occurred for the first time. PMID- 8243181 TI - [A prospective study of dynamic and epidemic factor of HBV infection in the army]. AB - In order to study the HBV infection in the army a total of 519 HBV susceptibles were observed prospectively for five years. The results showed that the cumulative infection rate of HBV was 16.12/1000 person-years and the latent infection was in the majority. There are many factors that could affect army population, the rate of HBsAg positive and the contact history with HB patients were indirect correlation with the rate of new HBV infection. Therefore it is an urgent matter to manage asymptomatic HBsAg carriers and HB patients in the army. PMID- 8243182 TI - [A path analysis of risk factors in relation to coronary heart disease]. AB - The risk factors for CHD were analyzed with path analysis in this paper. The results suggested that in order of size the direct factors affecting CHD were physical activities, quantity of smoking, intensity score of life events, family history of CHD, social supports, systolic blood pressure, triglyceridemia, type B behaviour and the indirect factors were diastolic blood pressure, cholesterolemia, family position, weight index, state of falling asleep, and siesta. The relations among factors and the degree of every factor's influence on CHD were analyzed according to the path figure and the size of total effect of every factor. The structural model of risk factors for CHD was built. PMID- 8243184 TI - [The development and extensive application of vitreous body surgery]. PMID- 8243183 TI - [Research on quality of life]. PMID- 8243185 TI - [The use of domestic silicone oil in the treatment of complicated retinal detachment]. AB - Vitrectomy and domestic silicone oil were used in the treatment of 42 eyes with complicated retinal detachment, and the retina was reattached in 34 cases or a rate of 80%, which dropped to 54% in 4-24 months postoperative. Among 7 eyes with successful retinal attachment and silicone oil removal, 1 eye developed recurrent retinal detachment. The therapeutic effect of silicone oil and operative complications were discussed. Removal of intravitreal silicone oil 3-6 months after injection was necessary to avoid adverse consequences. The study indicated that pars plana vitrectomy with silicone oil injection was an effective procedure in the management of complicated retinal detachment. PMID- 8243186 TI - [Perfluoropropane for the treatment of retinal detachment]. AB - Perfluoropropane (C3F8) is an ideal intravitreal gas tamponade in retinal detachment surgery and vitrectomy, as it is nontoxic, expansive and slowly absorbable. Domestic C3F8 of 99.7% purity was produced and used in 1990. An initial series of 120 cases received the pure or air-mixed C3F8 injection into the vitreal cavity during the retinal reattachment procedure. Application of the gas simplified the treatment of macular holes with retinal detachment, facilitated the replacement of everted retinal flaps of giant tears, and flattened the retina with vitrectomy. PMID- 8243187 TI - [Micro-vitreoretinal surgery for the treatment of complicated retinal detachment]. AB - Micro-vitreoretinal surgery combined with scleral buckling and intraocular tamponade were used in the treatment of 108 cases of complicated retinal detachment, including 5 cases with turbid media, 14 cases with traction bands, 53 cases with severe proliferative vitreoretinopathy, 13 cases with giant tears, and 23 cases with posterior retinal breaks. After a follow-up of 3-28 months, 69 cases (63.9%) achieved total retinal reattachment. Visual acuity was improved in 69 cases, unchanged in 24 cases, and decreased in 15 cases, 47 cases obtained visual acuities of over 0.05. The surgical procedures in different groups were discussed. PMID- 8243188 TI - [Vitreo-retinal surgery for complicated retinal detachment]. AB - 93 eyes (93 patients) of complicated retinal detachment were treated with vitreo retinal surgery. Among the series, 75 eyes were rhegmatogenous with PVR C3-D3 in 66 eyes (88.0%), while the remaining 18 eyes were traction induced. None of the cases had giant tears or complicating diabetes. On discharge from the hospital, the operation was effective in 62 cases (66.7%), in whom the retina was totally reattached or only a small amount of subretinal fluid remained. In a group of 40 eyes where the inert gas SF6 was used, the operation was effective in 30 cases (75.0%). 41 cases were followed up postoperatively for over 3 months, averaging 13.7 months, to find the operative results stable in 33 eyes (80.5%), with the visual acuity improved in 22 cases (66.7%), unchanged in 9 cases (27.3%), and decreased in 2 cases (6.0%). The operative procedures, the peeling of pre-retinal membrane, the effect of PVR severity on the operative results, and the promotion of operative efficacy by application of wide encircling buckle and inert gas tamponade were discussed. PMID- 8243189 TI - [Observations on vitrectomy with silicone oil injection]. AB - Silicone oils were injected in 37 eyes of 37 patients among 95 patients performed vitrectomy during 1988-1990. The indication was complicated retinal detachment except for 2 cases of diabetic proliferative retinopathy and vitreous hemorrhage. In 29 (78.4%) of the 37 eyes, the retina was totally reattached, and in 7 eyes partially reattached that did not regain ambulatory vision. In 22.7% of the eyes successfully operated, cataracts developed with silicone oil 1,000mPa.s while transient hypertension occurred in 27.3% with silicone oil 5,000mPa.s. Vitrectomy with silicone oil filling may help some retinal detachments that are refractory to conventional retinal and vitreous operations; however, due to possible complications, the use of silicone oil should be in limited cases and removed after retinal reattachment. PMID- 8243190 TI - [The operative use of perfluorodecalin in severe proliferative vitreoretinopathy]. AB - Perfluorocarbon liquids can be used as an operative hydrodynamic tool during vitreous surgery. The high specific gravity exerts a pressure on the retina to open up the retinal funnel, to displace the subretinal fluid and to stabilize the posterior retina for membrane peeling, endo-laser photocoagulation, and to release anterior traction. The authors reviewed 27 patients of a variety of severe proliferative vitreo-retinopathies (PVR) treated with the perfluorocarbon procedure, including idiopathic PVR in 9 cases, traumatic PVR in 7 cases, aphakic or pseudophakic PVR in 5 cases, intraocular inflammatory PVR in 4 cases and other forms in 2 cases. The success rate was 81.5% and the final visual acuity ranged from 0.05 to 0.5 in 50.0% of the patients that attained retinal reattachment. The retina failed to reattach in patients with rupture of the eyeball or advanced proliferative diabetic retinopathy. The surgical techniques and management of operative complications were discussed. PMID- 8243191 TI - [Treatment of rhegmatogenous retinal detachment complicated by vitreous hemorrhage]. AB - 8 cases of rhegmatogenous retinal detachment associated with vitreous hemorrhage were treated by first removing the blood through vitrectomy followed by routine retina reattachment procedures, such as searching for retinal breaks with indirect ophthalmoscopy and scleral buckling or encircling. Among 6 cases followed up, only 1 cases relapsed due to PVR traction. The points of note during operation were mentioned and the relationship between retinal breaks with vitreous hemorrhage and posterior vitreous detachment discussed. PMID- 8243192 TI - [Marfan's syndrome with retinal detachment]. AB - 14 patients (16 eyes) of Marfan's syndrome are reviewed with regard to retinal detachment. The patients aged a mean 25.1 years; 9 cases were of high myopia, and lens dislocation was found in all. Multiple atrophic retinal holes were detected in 11 eyes (68.8%), and the retina was totally detached in 12 eyes (75.0%). Scleral encircling with external compression performed in 15 eyes achieved retinal reattachment in 14 eyes. The predisposing factors for retinal detachment, the selection of the mode of surgery, the management of the pupil, and the prognosis are discussed. PMID- 8243193 TI - [Quantitative study of diabetic retinopathy by computerized image analysis]. AB - Composite fluorescein angiographic pictures of 66 eyes with diabetic retinopathy and nonperfusion areas were studied by computerized image analysis, and the ratio of the nonperfusion area to the disc area was calculated. There were 49 eyes with neovascularization (NV), in which the mean ratio +/- standard error was 46.33 +/- 7.01 (8.15 to 298.26), and the corresponding value in the other 17 eyes without NV was 6.90 +/- 1.31 (2.20 to 21.37), the difference being very significant. When the ratios were under 10, from 10 to 30, or over 30, the percentages of eyes manifesting NV were 13.3%, 81.8% and 100% respectively. In the 49 eyes with NV, the majority of the total 267 neovascular clusters were distributed 4-6 DD away from the optic disc, which itself was involved only in 18 eyes. PMID- 8243194 TI - [An experimental study of cyclocryocautery and its application on glaucoma patients]. AB - Cyclocryocautery of different time duration was done on 52 rabbit eyes with the Syncron Optikon CO2 cryocoagulator. Postoperative studies of the ciliary body by light and electron microscopy and enzymatic histochemical analysis at various intervals revealed morphologic and metabolic changes in the epithelial cells that resulted in diminished aqueous humor formation and lowered intraocular pressure. 50 cases of glaucoma patients (54 eyes) were treated with cyclocryotherapy and followed up over 6 months, with a success rate of 72% in the advanced refractory patients and the pain was relieved in 85%. 8 of the ineffective eyes were performed additional cyclocryotherapy under a scleral flap, with good results in 5 eyes. PMID- 8243195 TI - [Refractive evaluation of donor corneas from the newborn in aphakic penetrating keratoplasty]. AB - Donor corneas from newborns aged 1-28 days were used in 30 cases of aphakic penetrating keratoplasty with good refractive results. After an average postoperative follow-up of 14.8 months, the mean visual acuity was 0.3 s.c. and 0.6 c.c; the mean keratometric corneal refraction was +53.04 DS (about 10.00 DS higher than that of the contralateral eye), and the mean corneal astigmatism was 2.84 DC. The postoperative specular corneal endothelial count averaged 3,324 cells/mm2 in 14 eyes. PMID- 8243196 TI - [Observations on 108 normal ophthalmic arteries in 80 patients by digital subtraction angiography]. AB - By digital subtraction angiography, the authors made observations on 108 normal ophthalmic arteries in 80 patients with regard to the origins, courses, branching, morphology and anastomoses of the vessels. The merits and demerits of DSA in observing the ophthalmic artery were discussed. PMID- 8243197 TI - [An experimental study of ocular contusion in rabbits: the changes in electroretinography]. AB - In rabbits, the amplitude of a-waves and b-waves in the ERG declined following ocular contusion; however, the oscillatory potentials did not change markedly. In the anisodine therapeutic group, the amplitude of a-waves recovered higher than did that in the control group at the 2nd and 4th week post-traumatic, while all a waves recovered at the 6th week in both groups. The amplitude of b-waves was restored to normal in 72 hours for all eyes. Since the a-wave was sensitive to contusion, it monitored the severity of damage and objectively evaluated the therapeutic effect of treatment. PMID- 8243198 TI - [A study on RBC immunofunction in experimental traumatic uveitis]. AB - Examination of the RBC immunofunction in rabbit experimental traumatic uveitis with Guo's method of complement-labelled yeast revealed that the percentage of RBC C3b receptor rosette and the activity of red cell immune adherence (RCIA) enhancing factor in sera were markedly lower, while the percentage of RBC surface immune complex rosette, the activity of the RCIA-inhibiting factor and the CIC in sera were higher than those in the normal controls, indicating that the lowered RBC immunofunction was involved in pathogenesis of the experimental traumatic uveitis. PMID- 8243199 TI - [Analysis of 24 cases of misdiagnosed orbital tumors with visual impairment as the presenting symptom]. AB - 24 cases of orbital tumors misdiagnosed as optic neuritis or optic atrophy were located at the orbital apex or on the optic nerve, comprising meningioma 9 cases, cavernous hemangioma 5 cases, neurilemmoma 5 cases, neogrowths form the nasal cavity or paranasal sinuses 3 cases, leiomyoma 1 case and hemangiopericytoma 1 case. The authors analyzed the causes of CT examination in early differential diagnosis. PMID- 8243200 TI - Guideline for flow cytometric immunophenotyping: a report from the National Institute of Allergy and Infectious Diseases, Division of AIDS. PMID- 8243201 TI - Statistical methods in interphase cytogenetics: an experimental approach. AB - In situ hybridization (ISH) techniques on interphase cells, or interphase cytogenetics, have powerful potential clinical and biological applications, such as detection of minimal residual disease, early relapse, and the study of clonal evolution and expansion in neoplasia. Much attention has been paid to issues related to ISH data acquisition, i.e., the numbers, colors, intensities, and spatial relationships of hybridization signals. The methodology concerning data analysis, which is of prime importance for clinical applications, however, is less well investigated. We have studied the latter for the detection of small monosomic and trisomic cell populations using various mixtures of human female and male cells. With a chromosome X specific probe, the male cells stimulated monosomic subpopulations of 0, 1, 5, 10, 50, 90, 95, 99, and 100%. Analogously, when a (7 + Y) specific probe combination was used, containing a mixture of chromosome No. 7 and Y-specific DNA, the male cells simulated trisomic cell populations. Probes specific for chromosomes Nos. 1, 7, 8, and 9 were used for estimation of ISH artifacts. Three statistical tests, the Kolmogorov-Smirnov test, the multiple-proportion test, and the z'-max test, were applied to the empirical data using the control data as a reference for ISH artifacts. The Kolmogorov-Smirnov test was found to be inferior for discrimination of small monosomic or trisomic cell populations. The other two tests showed that when 400 cells were evaluated, and using selected control probes, monosomy X could be detected at a frequency of 5% aberrant cells, and trisomy 7 + Y at a frequency of 1%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243202 TI - Three-dimensional visualization of multi-channel volume data: the amSFP algorithm. AB - In this paper we present a three-dimensional visualization technique for multi channel volume data. The technique simulates the physical process of fluorescence, hence its name: achromatic multi-channel simulated fluorescent process (amSFP). The data set is simulated as 3D distribution of different fluorescent dyes, where each channel is represented by a particular type of dye. Apart from the spatial density map, no additional characteristics about the data set have to be defined; no image segmentation is needed prior to visualization. The degree of interaction among the channels in the fluorescence process can be adapted to optimally render specific structures in the image. 3D multi-channel data can be obtained by a three-dimensional imaging device that is able to measure a number of physical quantities at a given location within a specimen. The fluorescence principle, the algorithm, and its implementation are presented. We have used the technique to investigate the relative spatial arrangement of blood vessels and astrocytes in the cat retina. The two components have been stained with different fluorescence dyes and recorded in a confocal light microscope to form a two-channel 3D data set. PMID- 8243203 TI - Concurrent analysis of intracellular glutathione content and gap junctional intercellular communication. AB - The potential for performing dual analysis of intracellular glutathione levels and assessment of gap junctional intercellular communication with thiol-specific fluorescent probes in anchored cells was evaluated. Gap junction-mediated diffusion of monochlorobimane and 5-chloromethylfluorescein diacetate following intracellular loading and conjugation with glutathione was compared with 5 carboxyfluorescein diacetate (which is routinely used in laser cytometry to monitor intercellular communication) by means of fluorescence recovery after photobleaching using a variety of communication-competent and communication incompetent cells. The rate of diffusion of fluorescence among communication competent cells was inversely proportional to the size of the fluorescent probe employed. The thiol-specific probes were also employed to monitor depletion and synthesis of glutathione following treatments to inhibit glutathione synthesis or consume glutathione by adduct formation. Analysis of gap junctional intercellular communication following glutathione depletion revealed a direct correlation between glutathione levels and intercellular communication. These studies support the utility of the thiol-specific probes to monitor the respective role of cellular glutathione and intercellular communication in the mechanisms of cellular injury. PMID- 8243204 TI - Phenotype analysis of cycling and postcycling thymocytes: evaluation of detection methods for BrdUrd and surface proteins. AB - We present a comparison of two different methods for simultaneous detection of bromodeoxyuridine and cell surface markers. Both methods use enzymatic generation of single-strand DNA with nuclease. The biological system used is the murine thymus, in which in vivo DNA synthetizing cells were labeled by injection of BrdUrd and analyzed at different time points after the nucleoside pulse. The surface proteins detected were CD4 and CD8 differentiation markers and the T-cell receptor. Extraction of DNA-associated proteins with 0.1N HCl and detergent is necessary for the action of EcoR1 and Exonuclease III, but this treatment destroys phycocyanins and induces cell aggregation, as shown using the doublet discrimination module. For DNAse I action, cells could be treated with paraformaldehyde and a low concentration of Tween 20, and this treatment was adequate for surface staining preservation (even with phycocyanins) and BrdUrd detection. Both methods were adequate for cell cycle studies, but only 7-amino actinomycin D could be used as total DNA dye after DNAse action, and good results needed long (48-72 h) incubation in the fixative-detergent mixture. The DNAse I method now allows three-color staining (two surface markers and Brd-Urd), analyzed in a one laser-cytometer for the study of the phenotype of cycling cells, and of their progeny, in vivo and in cell cultures. It also allows the quantitative analysis of cell surface receptor densities in conditions similar to fresh cells. PMID- 8243205 TI - Nucleated cells response to protein electroinsertion. AB - Application of an electrical pulse field at a strength slightly below the value required for electroporation to a suspension of red blood cells in the presence of membrane xenoproteins leads to the insertion of those proteins in the erythrocyte plasma membrane. This observation is extended to nucleated cells. In the presence of glycophorin A, application of such pulses leads to the insertion of 10(4)-10(5) molecules of glycophorin A per cell in CEM-CM3, Hela S3, and bovine CD8+ T cells. Electroinserted glycophorin A is detected by flow cytometry using anti-glycophorin monoclonal antibodies. The survival of the cells subjected to electroinsertion was 55% for CEM-CM3 cells, 69% for Hela S3 cells, and 65% for CD8+ T cells. Cells cultured after electroinsertion lost the electroinserted glycophorin A, with two different rates, by a temperature and cell type-dependent mechanism. During the first 2 h after electroinsertion, the CD8+ T cells lost 12.5% of the inserted glycophorin A per h, the CEM-CM3 cells lost 7.7% per h, whereas the Hela S3 cells lost only 0.8% of the inserted protein per h. After 2 h, the rate increased substantially, to 41.7% per h for the CD8+ T cells, 13.5% for the CEM-CM3 cells, and 8.9% for the Hela S3 cells. Cytochalasin D efficiently inhibited the disappearance of electroinserted glycophorin A during the first 2 h after electroinsertion only. PMID- 8243206 TI - Analysis of lymphocyte activation and proliferation by video microscopy and digital imaging. AB - Video microscopy and digital imaging were used as a noninvasive method to quantitatively analyze lymphocyte activation and proliferation. This method takes advantage of the fact that upon activation lymphocytes blast and become significantly larger before proliferating. The mean cell sizes of T lymphocytes in an activation kinetics assay were measured by digital image analysis and compared to [3H]-thymidine incorporation of cells under the same treatment. An increase in cell size was observed before [3H]-thymidine incorporation; therefore the digital imaging assay is more sensitive in determining the earliest time point of activation. Also, the digital imaging assay was comparable to the [3H] thymidine incorporation assay in providing information about the extent and rates of T lymphocyte proliferation. Cellular DNA was stained with propidium iodide to show that the larger blasting cells in the population of activated T lymphocytes were indeed the cells that accounted for the increase in DNA synthesis and thus an increase in cell size can be correlated with activation. PMID- 8243207 TI - Characterization of the differentiation of human colorectal cancer cell lines by means of Voronoi diagrams. AB - This paper describes differentiation in terms of population dynamics through the medium of Voronoi paving which enables (via digital cell image analysis) the structure of human LOVO and HCT-15 colorectal neoplastic cell colonies growing on histological slides to be characterized. Two other tests were also used, i.e., the colorimetric MTT assay that enables the cell growth level to be determined, and a test allowing the assessment of the proliferation index, i.e., the percentage of cells in the S phase of the cell cycle. The results show that these colorectal neoplastic cells exhibited a comparatively high level of organisation in terms of the topographical distribution of nuclei within the clones when the cells were cultivated in media containing even small amounts of fetal calf serum. On the other hand, certain chemically defined media completely overturned this "pseudo-tissular" architecture. Furthermore, the colorectal cells growing in media including fetal calf serum exhibited relatively large and dense clones, undergoing an increase in the density of these clones when hormones were added to the culture medium and, concomitantly, a decrease in their proliferation. In contrast, the cells growing in chemically defined media generally exhibited smaller clones whose cell proliferation was paradoxically greater than that of the cells referred to above. This seems to bring out the importance of the part played by the cell loss factor in this cell population dynamic. PMID- 8243208 TI - Preparation of cell nuclei from fresh tissues for high-quality DNA flow cytometry. AB - An easy method for preparation of bare cell nuclei from fresh solid tissues for DNA flow cytometry is described. Pieces of up to 2 x 2 x 2 mm3 size from fresh tissues were fixed in formalin. After removal of formalin by washing with ethanol and rehydration with tap water, the tissue pieces were incubated with subtilisin Carlsberg (pronase, Sigma protease XXIV) and then stained directly with DAPI. Staining with ethidium bromide gave unsatisfactory results. Neither mechanical disaggregation nor centrifugation were used. The resulting cell nucleus suspensions had extremely low frequencies of debris particles and of clumped cell nuclei. A good yield, a minimized loss, and a good stainability of cell nuclei were obtained. The applicability of the method was exemplified by the analysis of biopsies from the colon-rectum in patients with ulcerative colitis and of biopsies from the bladder in patients with bladder cancer and compared to the standard method of this laboratory, which uses mechanical disaggregation, ethanol fixation, pepsin treatment, and staining with ethidium bromide. The formalin subtilisin Carlsberg technique resulted in good agreement of ploidy measurements compared to the standard method, a higher number of evaluable histograms, an improved detectability of aneuploid cell populations, and an improved accuracy of the S- and G2-phase analysis, particularly in samples with low proliferation. The method also makes it possible to use long-term storage and to transport samples by post. PMID- 8243209 TI - Preferential loss of abnormal prostate carcinoma cells by collagenase treatment. AB - Effects of two different methods of tumor disaggregation on flow cytometric ploidy distribution and intact cell yield were investigated. Either mechanical disaggregation or collagenase digestion was applied to 35 prostate tumor specimens. Seven collagenase-treated samples failed to yield any intact cells, whereas with mechanical disaggregation in all cases a sufficient number of intact cells were obtained. No differences in the FCM ploidy distribution of tumors with a DNA diploid stemline were observed comparing both techniques. In DNA aneuploid tumors, however, collagenase treatment had an adverse effect on the abnormal cell populations. In 14/17 of such tumors, the abnormal cell populations were significantly reduced; in eight of these the percentage of DNA aneuploid cells declined even below the minimum percentage (10%) that was defined for DNA aneuploidy. Since collagenase is a widely used enzyme for tissue disaggregation, especially in tumor cytogenetics, the presented data will have consequences for the interpretation of results obtained by methods involving the use of this enzyme. PMID- 8243210 TI - Rapid flow cytometric antibiotic susceptibility assay for Staphylococcus aureus. AB - Conventional techniques for antibiotic susceptibility testing usually require 24 h or more to produce accurate results. This long waiting period to demonstrate antibiotic action is necessary because such tests rely on growth (or the lack of it), when the microorganisms are incubated in the presence of the antibiotic. In an effort to improve antibiotic susceptibility testing, we developed a flow cytometric technique for Staphylococcus aureus in which antibiotic action is determined by monitoring drug-induced membrane potential changes, using the potential-sensitive dye 3,3'-dipentyloxacarbocyanine iodide. Three ATCC reference strains of S. aureus and 13 unknown strains of the same microorganism were tested for susceptibility to penicillin G and oxacillin. Our results indicate that susceptibility of S. aureus to these antibiotics can be measured reliably at 90 min after addition of the antibiotic, and the results are comparable to those obtained with conventional susceptibility tests. PMID- 8243211 TI - Beam-shaping optics for a slit-scan flow cytometer. AB - Two sets of beam-shaping optical systems for slit-scan flow cytometry were made for 360 nm and 488 nm wavelength argon-ion lasers. A thin, ribbonlike laser beam was formed by compressing the zero-order component of Fraunhofer diffraction patterns caused by a slit vertically long and horizontally narrow by means of an objective with a numerical aperture of 0.5. The objectives were designed to include a jet stream that performed as a cylindrical lens. The characteristic values of the optical systems and lens alignments of the systems are described in this work. The beam size at focus was 0.95 micron in thinness and 85.0 microns in width on design values. The actual beam thinness for the UV laser was estimated to be 1.3 microns, based on shapes of pulses generated when fluorescence microspheres of diameter 1.0 microns passed across the beam. The usefulness of the optical systems was also proved by its ability to measure HeLa chromosomes. PMID- 8243212 TI - Flow cytometric assay of pinocytosis: correlation with membrane ruffling and metastatic potential in the Dunning R-3327 rat prostatic adenocarcinoma model. AB - Membrane ruffling has been associated with neoplastic transformation, Harvey ras expression, and metastatic capability. In the Dunning R-3327 rat prostatic adenocarcinoma model, membrane ruffling graded visually upon live cultured cells filmed by time-lapse video-microscopy has distinguished sublines of high and low metastatic potential. Fluid-phase pinocytosis is a constitutive, noninducible internalization of medium by cell membrane. Fluid phase pinocytosis may be measured flow cytometrically by cellular uptake of fluorescein-labelled medium constituents. The optimum conditions for a flow cytometric assay of pinocytosis were determined using AT-2 subline that has an intermediate degree of membrane ruffling. The optimum dextran concentration was selected from the midpoint of the linear portion of the dose-response (0.01-10.00 mg/ml) curve, whereas the optimum incubation time was determined from a time-course (1-405 min.) curve study. Cultured cells from 6 Dunning sublines incubated with 1.0 mg/ml of fluorescein labelled dextran for 90 min were washed, fixed, and the fluorescence of 10,000 cells studied by flow cytometry. For each subline, dextran fluorescence was measured in four independent experiments. Pinocytosis failed to distinguish sublines of high (AT-3 63.5 +/- standard error 4.1 mean channel number, MAT-LyLu 63.2 +/- 6.3, MAT-Lu 64.3 +/- 5.6) and low (G 33.5 +/- 1.2, AT-1 63.5 +/- 4.1, AT 2 58.4 +/- 3.6) (rank p = 0.38) metastatic potential but correlated strongly with visually graded membrane ruffling (r = 0.95, p = 0.003). Pinocytosis assayed by flow cytometry reflects membrane ruffling observed visually and thus flow cytometric assays may facilitate study of membrane activity. PMID- 8243213 TI - Development of a shape vector that identifies critical forms assumed by human polymorphonuclear neutrophils during chemotaxis. AB - Human polymorphonuclear neutrophils undergo characteristic shape changes that are critical to their ability to move and ingest their targets. We present here the construction of a simple shape vector, calculated from the coordinates of the cell perimeter, that can identify critical forms that a neutrophil assumes during the course of ameboid movement. The vector can be used to find neutrophils of a specific shape from the image analyzer data produced during a typical neutrophil tracking experiment. PMID- 8243214 TI - Positioning during hemodynamic monitoring: evaluating the research. AB - Can pulmonary artery or right atrial pressures be obtained accurately when patients are in lateral positions? This review of the published research says probably not. PMID- 8243215 TI - Venipuncture vs. arterial catheter activated partial thromboplastin times in heparinized patients. AB - The use of arterial or venous access for obtaining a blood sample to determine the activated partial thromboplastin time (aPTT) remains controversial. The findings of this study indicate that only access site affects aPTT. Findings such as these can be used by critical care nurses to develop unit-based standards. PMID- 8243216 TI - SCIWORA syndrome: nursing assessment. AB - Spinal Cord Injury Without Radiographic Abnormality (SCIWORA) is a commonly misdiagnosed injury in children and adolescents. Failure to recognize SCIWORA can lead to additional injury. The critical care nurse recognizes the risk of SCIWORA, conducts a complete neurologic assessment on children experiencing cervical trauma, and provides immediate immobilization to prevent further neurologic complications. These pediatric neurosurgical and trauma nurse authors describe the nurse's role and include a SCIWORA case study in this article. PMID- 8243217 TI - Nursing strategies for patients with silent myocardial ischemia. AB - While angina pectoris has been thought to be a reliable indicator of myocardial ischemia, it is now known that many ischemic episodes are painless or "silent." Critical care nurses play an essential role in detecting this silent disease as well as providing acute nursing care. PMID- 8243218 TI - Increasing your decision-making role in ethical situations. AB - Critical care nurses are increasingly involved in situations of ethical conflict; yet, they are often left out of consequential decision making. This article discusses ways critical care nurses can become more deeply involved in actual decision making in ethical situations. PMID- 8243219 TI - Using a model to assess family satisfaction. AB - Family satisfaction with nursing care is difficult to evaluate because there are many factors which influence it, especially in critical care areas. These authors present a model for evaluating the primary factors which affect family satisfaction with critical care nursing. PMID- 8243220 TI - Issues in biomedical ethics. AB - Bioethics is the discipline of ethics dealing with moral problems arising in the practice of medicine and the pursuit of biomedical research. Physicians may confront ethical dilemmas regularly in their individual relationships with patients and in institutional and societal decisions on health care policy. Ethical problem solving requires the application of certain ethical rules and principles to specific situations. Although ethical theories differ, certain ethical rules and principles appear consistently. These include nonmaleficence, beneficence, respect for individual autonomy, confidentiality, and justice. This article discusses some of the ethical issues that arise in clinical practice, including informed consent, do-not-resuscitate orders, noninitiation and termination of medical therapy, genetic intervention, allocation of scarce health resources, and infection with the human immunodeficiency virus (HIV). Some of these problems require ethical analysis at the bedside; others require physician involvement on a broader level. Perspectives on the different ethical issues are presented; however, absolute answers to these ethical dilemmas are not provided. Interpretation of the ethical principles and the application of these principles to each clinical situation demands the thoughtful attention of the practitioner. PMID- 8243221 TI - [Treatment of systolic hypertension in the elderly]. PMID- 8243222 TI - [Evaluation of the ambulatory blood pressure monitoring]. PMID- 8243223 TI - [Systolic hypertension in the elderly: Chinese trial (Syst-China)--second interim report]. AB - Two thousand and one hundred thirty elderly isolated systolic hypertensive patients entered the multicenter, randomized, double-blind, placebo-controlled clinical trial (1110 in the active treatment group and 1020 on placebo). The differences of ages, height, weight, sex constituent and baseline blood pressure between active treatment and placebo groups yielded no significance. In the 3.2 years (median) followup, the systolic and diastolic blood pressure in the active treatment differed from those in the placebo and the former were lower than the latter (P < 0.05). The prevalence of side effects or symptoms were greater in the placebo group than in the active treatment group. The present trial is in progress and the final results will not be reported until the end of the trial. PMID- 8243224 TI - [The relationship between ambulatory blood pressure and left ventricular hypertrophy in essential hypertension]. AB - The relationship between ambulatory blood pressure (ABP) and left ventricular hypertrophy (LVH) were investigated in 137 consecutive subjects (54 healthy normotensives and 83 uncomplicated patients with essential hypertension) who underwent 24-hour noninvasive ABP monitoring and M-mode echocardiography. In the normotensive group, left ventricular mass index (LVMI) was not correlated with average daytime (6AM-10PM) or nighttime (10PM-6AM) systolic and diastolic blood pressure (SBP and DBP) except for age (r = 0.405, P < 0.01). In the hypertensive group, however, LVMIs were significantly correlated with average daytime SBP (r = 0.315, P < 0.01), nighttime SBP (r = 0.408, P < 0.01) and DBP (r = 0.304, P < 0.01). Furthermore, significant inverse correlations were found between LVMI and percentage of nocturnal reduction of daytime SBP (r = -0.393, P < 0.01) and DBP (r = -0.308, P < 0.01). The results suggest that the average levels of ABP, particularly, the level of ABP during night and the loss of circadian variation of blood pressure, would be responsible for the development of LVH. The control of blood pressure and restoration of circadian blood pressure variation might be necessary for the reversal of LVH in hypertensive patients. PMID- 8243225 TI - [Changes in plasma neuropeptides before and after clonidine in patients with essential hypertension]. AB - Plasma concentrations of beta-endorphin (beta-EP), leucine enkephalin (LEK), arginine vasopressin (AVP), neurotensin (NT), renin activity (PRA) and angiotensin II (AT-II) were determined before and after the treatment with clonidine in 117 patients with essential hypertension. Before the treatment, the patient group had lower levels of beta-EP and LEK (P < 0.001), higher levels of AVP, PRA and AT-II (P < 0.05-0.01), as compared with those in control group. After 14 days of the treatment, plasma levels of beta-EP, LEK increased significantly (P < 0.001), and correlated negatively with the decrease of the mean artery pressure (r = -0.369 and r = -0.441, respectively, P < 0.01). PRA and AT-II decreased significantly (P < 0.05, P < 0.01). Decrease of AVP level was also observed, but did not reach the statistical significance. NT did not change both before and after the treatment. These data suggest that beta-EP and LEK may be involved in pathogenesis of hypertension and in hypotensive action of clonidine. PMID- 8243226 TI - [Changes in regulatory peptides at different stages of essential hypertension and their clinical significance]. AB - The changes of regulatory peptides in 100 patients with essential hypertension at different stages were studied. Four kinds of peptides in serum were measured by RIA method. The results showed that: (1) neurotensin, P-substances, beta endorphin and leucine-enkephalin decreased significantly in the group (P < 0.01). (2) the levels of those parameters at different stages of hypertension were decreased in parallel with its severity and the difference between different stages is significant (P < 0.01). (3) norepinephrine showed negative correlation with four kinds of neuropeptides. (4) neurotensin, Beta-endorphin and leucine enkephalin increased significantly after capton (P < 0.01). The possible mechanism and its clinical significance of the changes of those regulatory peptides in hypertension patients were discussed. PMID- 8243227 TI - [The changes in plasma arginine-vasopressin in patients with essential hypertension and the correlation with patient's condition]. AB - The levels of plasma arginine-vasopressin (AVP) in 80 patients with essential hypertension were measured, and its impact on the disease and its clinical significance were studied. The results showed that: (1) The levels of plasma AVP in patients with essential hypertension were significantly higher than that in normotensive subjects (P < 0.001). It dropped to normal level after antihypertensive drugs. (2) The concentrations of plasma AVP in both hypertensive subjects and normotensive subjects were not correlated with age and sex (P < 0.05). (3) The concentration of plasma AVP in patients with essential hypertension was the highest in stage III, the lowest in stage I, and middle in stage II. (4) The levels of plasma AVP in patients with malignant hypertension were significantly higher than that in patients with benign hypertension (P < 0.05). A positive correlation was found between the levels of plasma AVP and blood pressure (r = 0.3398, P < 0.01). (5) The concentrations of plasma AVP in hypertensive subjects with ventricular hypertrophy were higher than that in hypertensive subjects with out ventricular hypertrophy (P < 0.05). (6) The concentrations of plasma AVP in hypertensive subjects with heart failure were significantly higher than that in hypertensive subjects with out heart failure (P < 0.001). The results suggest that AVP has a role in the pathogenesis of hypertension, hypertension complicated with ventricular hypertrophy and hypertension complicated with heart failure. The levels of plasma AVP may be viewed as an index of the patient's condition in hypertensive subjects. PMID- 8243228 TI - [Changes in the fibrinolytic activity in hypertension and acute cerebral infarction]. AB - The fibrinolytic activity was measured in 30 patients with essential hypertension and 15 patients with acute cerebral infarction. The content of plasminogen (PLG) and plasminogen activator inhibitor-1(PAI-1) were higher in the patients with essential hypertension than in the controls. The results suggested that the fibrinolytic activity had a potential tendency towards decreasing in the patients with essential hypertension. The fibrinolytic activity was lower in the stage of cerebral attack and was higher in the stationary stage of acute cerebral infarction, which showed that a change of the fibrinolytic activity in acute cerebral infarction was related to the clinical course. PMID- 8243229 TI - [Surgical treatment of secondary atrial septal defect associated with mitral valve disease]. AB - From March 1965 through August 1991, 703 patients with secondary atrial septal defect were treated surgically in our hospital. Among them, 32 cases (4.6%) were associated with mitral valve disease. The mitral valve lesion was repaired at the same time of closure of the atrial septal defect. The operative mortality was 6.3% and there was no late death. Follow-up study showed that heart function of the patients was improved obviously and most of them have resumed work or study. The clinical features, incidence, etiology, pathology and surgical techniques of secondary atrial septal defect associated with mitral valve disease were discussed. PMID- 8243230 TI - [Percutaneous transfemoral plug closure of patent ductus arteriosus in 100 cases]. AB - One hundred patients with patent ductus arterious (PDA) were treated successfully by percutaneous transfemoral plug closure since 1985. Age ranged from 5 to 48 years. Among them five had previous ligation of PDA, One was complicated with infective endocarditis, and in four pulmonary hypertension were found. The internal diameter shown in aortograph ranged from 0.26-1.0 cm. Primary success was accomplished after the first trial in 97 patients and second trial was necessary in three cases. However in one patient plug closure was given up after unsuccessful trial. The plug closure was carried out with domestic-made material and instrument in all cases. We adopted the modified Porstmann technique. We believe that the infrainguinal transfemoral approach is simple, and less traumatic. The indication of plug closure can be extended especially to residual or recanalization cases after ligation in selected cases. It may become the method of first choice for PDA. The "papillary type" of PDA is discussed and a specially designed and tailored plug and also a spontaneous flow-directed method were recommended. Frequent ventricular premature beats usually occurred while the catheters were being pushed through chordae tendinae of the tricuspid valve and the way of prevention was discussed. In case of dislodgement of plug, reestablishing the wire locus would be necessary. One to sixty-eight months follow up study after closure showed no sign of recanalization or residual shunt. PMID- 8243231 TI - [Percutaneous transluminal peripheral excimer-laser angioplasty: two successful cases report]. AB - The excimer-laser (XeC1) operating at 308 nm, ablates by means of photochemical mechanisms with minimal thermal injury. Two male patients, 53 years and 67 years, with symptomatic peripheral vascular disease underwent percutaneous transluminal excimer-laser angioplasty (PTPELA). Both of them experienced claudication at a short walking distance and had superficial femoral or initial segmental femoral occlusions from 4 cm to 7 cm in length. The laser emitted 120ns pulses at 20 Hz with delivering 16-18 mJ by 7F and 9F catheter, energy density was about 4mJ/mm. The irradiation time was 157 seconds and 185 seconds respectively. Successful recanalization occurred in our two cases, the residual stenosis were less than 20%. The acute outcome was good without any complications. PMID- 8243232 TI - [A study on the circadian variation of tissue-type plasminogen activator and its rapid inhibitor in patients with coronary heart disease]. AB - In our study, we measured resting tissue-type plasminogen activator(t-PA) activity and resting plasminogen activator inhibitor-1 (PAI-1) activity in 15 healthy men and in 20 patients (male) with coronary heart disease in the morning and evening. Resting t-PA activity was lower (P < 0.05) in controls and was significantly lower (P < 0.001) in patients in the morning compared with that in the evening. In contrast, PAI-1 activity was significantly higher (P < 0.001) in controls and was also significantly higher (P < 0.001) in patients in the morning compared with that in the evening. t-PA activity in patients compared with that in controls was significantly lower (P < 0.05) in the morning and the evening; PAI-1 activity in patients compared with that in controls was significantly higher (P < 0.001) in the morning and the evening. However, t-PA activity was significantly lower (P < 0.05) and PAI-1 activity was significantly higher (P < 0.05) in patients with myocardial infarction compared with anginal patients in the morning. Applying this clinical phenomenon that myocardial infarction and angina have often occurred in the morning, the relationship between time of onset of myocardial infarction and circadian variation of t-PA and PAI-1 was discussed. PMID- 8243233 TI - [An evaluation of right ventricular systolic performance in patients with cor pulmonale with two-dimensional echocardiography]. PMID- 8243234 TI - [Ras oncogene expression of left ventricle in hypertrophic rat heart and during induction by vasopressin and neosynephrine]. AB - Using molecular hybridization, left ventricular ras oncogene expression was examined in the hypertrophic heart of rat or during injection of vasopressin or alpha-receptor agonist neosynephrine. Results showed that ras oncogene expression of left ventricle could be potentiated by neosynephrine but not by vasopressin. Furthermore, expression of both nucleic oncogene myc and membrane oncogene ras were increased during chronic cardiac pressure overload, with the former occurring in early loading stage and the latter staying the whole loading stage. PMID- 8243235 TI - [An experimental study on the mechanism of antiatherosclerotic effect of nifedipine]. AB - Thirty male N-Z white rabbits were randomized into 3 groups (10 each). Normal control group (C) were fed regular chow. Atherosclerotic control group (CAS) were fed a chow containing 1% cholesterol. Nifedipine group (N) were fed 1% cholesterol chow and nifedipine 40 mg/animal/day. The experiment lasted for 12 weeks. Blood total cholesterol, HDL-C, MDA, 6-keto-PGF1 alpha and TXB2 were measured. After the sacrifice of the animals, electron probe microanalysis was employed to cryo-ultrathin sections of the aorta for measuring cytoplasmic calcium content in situ in the medial smooth muscle cells. The area of atherosclerotic plaques of the aortae was planimetried with computer. The results showed that MDA and TXB2 in group CAS were markedly higher than those in groups C and N (P < 0.01 or 0.05). The cytoplasmic calcium in group CAS was significantly higher than that in groups C and N (P < 0.01). The difference between groups C and N was not significant. The area of atherosclerotic lesions in group CAS was 44 +/- 8%, markedly larger than that in group N (13 +/- 5%, P < 0.01). The antiatherosclerotic effects of nifedipine may be mainly by a mechanism of inhibiting intracellular calcium overload. PMID- 8243236 TI - [The protective effect of different drugs on ischemic-reperfusion injury in experimental pig heart]. PMID- 8243237 TI - [Ambulatory blood pressure monitoring research in hypertension and its clinical use]. PMID- 8243238 TI - [Complications after coronary angiography and balloon dilatation]. AB - All major complications within four weeks after diagnostic cardiac catheterization (DCCA; n = 4,778) or percutaneous transluminal coronary angioplasty (PTCA; n = 3,073) were analysed prospectively in 7,851 patients (5,769 men, 2,082 women; mean age 59.4 [16-88] years). There were 116 such complications in 114 patients, 46 (0.9%) after DCCA, 70 (2.3%) after PTCA. 76% of all complications occurred during the first two days after the procedure: bleeding in 29 patients, false aneurysms in 15, arterial occlusions in 12, venous ones in three, cerebral ischaemia in 12, fulminant pulmonary emboli in three, cardiac complications in 27, and others in 15. There were five deaths in connection with a DCCA, 18 with PTCA. Complications were twice as common in women as in men (DCCA: 1.5% vs 0.7%, P < 0.05; PTCA: 4.2% vs 1.7%, P < 0.001). Complications after DCCA were more frequent in those aged over 55 years (1.2% vs 0.3%, P < 0.05); after PTCA they were more frequent in those aged over 65 years (4.1% vs 1.3%, P < 0.05). Other risk factors for complications were: peripheral arterial occlusive disease for arterial occlusion (P < 0.001), high heparin dosage with DCCA (P < 0.05), and additional venous puncture with DCCA (P < 0.01). PMID- 8243239 TI - [Polyostotic fibrous dysplasia of the spinal column and ribs]. AB - The chest radiograph of a 56-year-old man with pain over the entire region of back, chest and vertebral column revealed chunky swelling over the 7th right rib. Other radiographs and magnetic resonance imaging further showed an infiltrative process in the 12th thoracic and the 2nd as well as 4th lumbar vertebrae. Skeletal scintigraphy was highly suggestive of bone metastases. Results of laboratory tests were within normal limits. Partial rib resection was performed after failure to find the site of a primary tumour. The diagnosis of polyostotic fibrous dysplasia was made histologically. There was no evidence of malignancy and a rare additional endocrinopathy (McCune-Albright syndrome) was excluded. PMID- 8243240 TI - [Acute kidney failure caused by sulfadiazine stones. A complication of the therapy of toxoplasmosis in AIDS]. AB - A 45-year-old man with AIDS was treated for a recurrence of cerebral toxoplasmosis with sulphadiazine, 4 g, and pyrimethamine, 75 mg, daily. Owing to a lack of appetite and dysphagia he drank rather little water during the first week of treatment. On the 13th day after starting the drugs he had bilateral renal colics and renal failure was diagnosed (serum creatinine 3.8 mg/dl). Ultrasound examination demonstrated multiple stones with bilateral urinary retention. After parenteral fluid replacement, alkalization of the urine with sodium-potassium-hydrogen citrate and N-butylcopolamine a stone, consisting of sulphadiazine and acetylsulphadiazine, was passed after two days. Three days later the creatinine concentration was within normal limits, and in further two days the ultrasound picture was normal. It is pointed out that diarrhoea, fever or dysphagia often prevent sufficient fluid intake in AIDS patients. Satisfactory oral fluid intake and alkalization of urine is thus of great importance for avoiding complications during sulphadiazine treatment. PMID- 8243241 TI - [Vesicorenal reflux]. PMID- 8243242 TI - [The prevention of Pneumocystis carinii pneumonia in HIV infection]. PMID- 8243243 TI - [The sauna in arterial hypertension]. PMID- 8243244 TI - [Lymph drainage]. PMID- 8243245 TI - [A transit thrombus in the patent foramen ovale with pulmonary and paradoxical embolization]. PMID- 8243246 TI - [The treatment of multiply resistant pulmonary tuberculosis]. PMID- 8243247 TI - [Life habits and mortality]. PMID- 8243248 TI - [Immune activity and serum interleukin-2-receptor concentration in autoimmune thyroid diseases]. AB - Serum interleukin-2 receptor (IL-2R) concentrations were compared in 55 patients with thyrotoxicosis (14 men, 41 women, mean age 43.5 +/- 17 years), 18 patients with Hashimoto's thyroiditis (5 men, 13 women, mean age 47 +/- 15 years) and 28 healthy subjects (12 men, 16 women, mean age 30 +/- 10 years). The patients with thyrotoxicosis were divided into three groups depending on the activity or stage of the disease: 17 patients with florid untreated hyperthyroidism, 23 euthyroid patients receiving treatment with antithyroid drugs and 15 patients with thyrotoxicosis in remission after completing one year's antithyroid treatment. The patients with untreated thyrotoxicosis had significantly higher IL-2R values than the euthyroid patients receiving treatment or those in remission (207 +/- 112 vs 139 +/- 66 and 91 +/- 26 U/ml, P < 0.05 and P < 0.01). The IL-2R values of patients with thyrotoxicosis in remission were, however, significantly lower than those of the 28 healthy subjects (126 +/- 34 U/l; P < 0.01) or the euthyroid patients receiving treatment (P < 0.05). The 18 patients with Hashimoto's thyroiditis had significantly lower serum IL-2R values (70 +/- 39 U/ml) than the healthy controls. These data show that the serum IL-2R level depends on the state of thyroid metabolism and on the activity phase of the thyrotoxicosis. The low serum levels of IL-2R in patients with Hashimoto's thyroiditis could signify a genetically determined decrease in IL-2R production or might be linked with the destruction of thyroid tissue by the chronic autoimmune process. PMID- 8243249 TI - [Pulmonary valve endocarditis, bilateral deep vein thrombosis amd recurrent pulmonary emboli]. AB - A 57-year-old obese man with hypertensive heart disease and long-standing varicose veins developed recurrent bouts of fever, lung infiltrations, cardiac arrhythmias and increasing dyspnoea. On admission multiple lung emboli and bilateral deep vein thromboses were confirmed. The echocardiogram demonstrated floating vegetations on all three pulmonary valve cusps. Biochemical tests indicated an inflammatory constellation (ESR 62/105 mm, C-reactive protein 13.3 mg/dl), partial respiratory insufficiency (pO2 54.6 mm Hg; pCO2 29 mm Hg). Streptococcus bovis was grown from several blood cultures. Conservative treatment over several weeks, complicated by "drug fever", with penicillin G (10 mega IU four times daily) and gentamicin (80 mg twice daily intravenously), later vancomycin (500 mg four times daily intravenously), then roxithromycin (150 mg three times daily by mouth), as well as maintenance anticoagulation with heparin (800-1,200 IU/h intravenously, later 15,000 IU subcutaneously twice daily), followed by phenprocoumon, contained the disease. One year after onset of treatment a good functional results had been achieved. PMID- 8243250 TI - [Eosinophilic fasciitis (Shulman syndrome) as a rare differential diagnosis of sclerodermiform skin changes]. AB - For 3 months, a 70-year-old woman had been affected by painless, symmetrical swellings in the limbs, spreading proximally, causing hardening of the skin and impaired joint movements, but no fever. Her general condition and findings on physical examination were according to age. ESR was 22/42 mm and there was a thrombocytosis (533,000/microliters) and eosinophilia (16%, absolute 2,140/microliters). Radiological and ultrasound examinations were normal. Skin biopsy revealed marked oedema of the connective tissue septa and perivascular lymphohistiocytic infiltrations, which--together with the other findings- suggested eosinophilic fasciitis. She was treated with prednisone, 100 mg daily, reduced gradually to 8 mg daily, as well as cimetidine, 800 mg daily, as immunomodulator, and intensive physiotherapy. The swellings and eosinophilia regressed within 8 weeks, but the hardening and impaired mobility persisted. Maintenance glucocorticoids are being continued. PMID- 8243251 TI - [Immunosuppression]. PMID- 8243252 TI - [Neurologic paraneoplastic syndromes. Anti-Purkinje-cell-antibodies and anti neuronal-nuclear-antibodies as serologic markers]. PMID- 8243253 TI - Troponin T. PMID- 8243254 TI - [Molsidomine and bladder cancer: is there a connection?]. PMID- 8243255 TI - [Acute respiratory failure in tropical malaria in pregnancy]. PMID- 8243256 TI - [Acute respiratory failure in tropical malaria in pregnancy]. PMID- 8243257 TI - [Acute respiratory failure in tropical malaria in pregnancy]. PMID- 8243258 TI - Estrous cycle stage-dependent effects of neuropeptide-Y on luteinizing hormone (LH)-releasing hormone-stimulated LH and follicle-stimulating hormone secretion from anterior pituitary fragments in vitro. AB - We recently demonstrated that neuropeptide Y (NPY) potentiates LH-releasing hormone (LHRH)-stimulated LH secretion in vivo, and that these actions of NPY are exerted only under the endocrine conditions leading to preovulatory LH surges, viz. the proestrous or estrogen- and progesterone-primed ovariectomized animal. The present experiments tested the hypothesis that NPY's facilitatory actions are exerted directly at the level of the anterior pituitary gland and depend on in vivo exposure of gonadotropes to the preovulatory endocrine milieu. Animals were killed at 1600 h on either metestrus or proestrus. Anterior pituitary glands (APs) were rapidly removed, cut into eighths, and placed into perifusion chambers. APs were perifused with medium 199 for a total of 8 h, and perfusate samples were collected every 5 min. After 30 min of equilibration, APs received hourly pulses of LHRH alone (10(-8) M), NPY alone (10(-6) M), or LHRH plus NPY. Basal runs consisted of perifusion with medium 199 for 8 h with no peptide treatments. Calculations of total hourly LH and FSH responses revealed that whereas LHRH significantly stimulated gonadotropin secretion from both metestrous and proestrous pituitaries, NPY significantly enhanced LHRH-stimulated LH and FSH secretion only from proestrous pituitaries, i.e. from tissue exposed to the endogenous endocrine milieu under which preovulatory gonadotropin surges are generated. NPY had no facilitatory effect on LHRH-induced gonadotropin secretion from metestrous APs. These results are consistent with our previous in vivo findings and demonstrate that the facilitatory actions of NPY on LHRH-stimulated gonadotropin secretion in vitro are limited to the endocrine conditions under which preovulatory gonadotropin surges are generated. PMID- 8243259 TI - RU486 administration blocks neuropeptide Y potentiation of luteinizing hormone (LH)-releasing hormone-induced LH surges in proestrous rats. AB - We previously demonstrated that NPY potentiates LHRH-induced LH secretion specifically under endocrine conditions in which preovulatory LH surges are generated. The present study was designed to test the hypothesis that NPY's facilitatory actions are dependent upon preovulatory progesterone secretion. In Exp 1, female rats were fitted with atrial catheters on diestrus. On proestrus, hourly blood samples were collected from 1100-2100 h. At 1230 h, rats received a sc injection of the progesterone receptor antagonist RU486 (6 mg/kg BW) or oil. At 1330 h, rats received pentobarbital (40 mg/kg BW), to block hypothalamic LHRH release, or saline. Every 30 min from 1400-1800 h, pentobarbital-treated rats received iv pulses of LHRH (15 ng/pulse) or saline along with concurrent pulses of NPY (5 micrograms/pulse), or saline. In Exp 2, rats received jugular catheters on diestrus, but were sampled every hour throughout the morning (0700-1600 h), rather than the afternoon, of proestrus. In these morning groups, pentobarbital was injected at 0830 h, and peptides (LHRH or combined LHRH and NPY solutions) were administered as pulses at 30-min intervals between 0900-1300 h. Results from Exp 1 were as follows: administration of RU486 to rats given an ip injection of vehicle at 1330 h and pulses of saline from 1400-1800 h completely blocked the endogenous LH surge. In oil-treated pentobarbital-blocked rats, concurrent administration of NPY with LHRH significantly (P < 0.01) potentiated the ability of LHRH to restore LH surges. However, NPY was without any potentiating effects in animals pretreated with RU486 at 1230 h. RU486 also attenuated the ability of LHRH alone to restore LH surges in pentobarbital-blocked rats. In Exp 2, NPY was without effect on LHRH-induced LH secretion during the morning hours of proestrus. Our results demonstrate that 1) NPY facilitates LHRH-induced LH surges on the afternoon of proestrus; 2) presumptive progesterone receptor blockade by RU486 completely prevents NPY's potentiating effects; and 3) NPY is without effect on the morning of proestrus, before the afternoon surge of progesterone. These findings are entirely consistent with the idea that one function of preovulatory progesterone secretion is to up-regulate pituitary sensitivity to the facilitatory actions of NPY. It is hypothesized that these actions of progesterone together with an increase in NPY neurosecretion mediate the acute increase in pituitary sensitivity to LHRH that occurs just before the LH surge. PMID- 8243260 TI - The chronic intracerebroventricular infusion of interleukin-1 beta alters the activity of the hypothalamic-pituitary-gonadal axis of cycling rats. I. Effect on LHRH and gonadotropin biosynthesis and secretion. AB - We have previously reported that the acute injection of interleukin-1 beta (IL-1 beta) into the brain ventricles of intact female rats promptly decreases LHRH release and inhibits gene expression of this peptide in the medial preoptic area (MPOA). The present studies were therefore designed to determine whether continuous exposure to the cytokine would disrupt the estrous cycle. IL-1 beta was injected intracerebroventricularly for 4-6 days at a rate of 4 ng/h. Daily vaginal smears were obtained to follow the cycle; pituitary LH and FSH secretion were measured at regular intervals. Steady state levels of LH and FSH messenger RNA (mRNA) in the pituitary, and LHRH gene expression in the MPOA, were measured at the end of the treatment. Infusion of IL-1 beta caused a total disruption of the estrous cycle, characterized by persistent smears indicative of the diestrus stage. When compared to animals treated with the vehicle, rats infused with IL-1 beta showed a significant decrease in circulating LH concentrations, which was accompanied by lowered mRNA levels in the pituitary. This statistical difference (P < 0.01) persisted even when treated rats were compared to control in a similar stage of the cycle (i.e. diestrus). Plasma FSH levels remained low at all times after IL-1 beta infusion but showed the expected cyclic changes in control animals. At the end of treatment, LHRH gene expression was also markedly suppressed in LHRH neurons distributed between the rostral preoptic area/organum vasculosum of the lamina terminalis and the MPOA of these animals. These results indicate that prolonged infusion of IL-1 beta into brain ventricles disrupts the estrous cycle, an event accompanied by decreased biosynthesis/release of LHRH and gonadotropins. We report in a related study that IL-1-treated rats also show increased plasma progesterone levels. However, it is improbable that this change was responsible for the interruption of the cycle described here; indeed we have previously observed that the central administration of IL-1 beta to intact rats resulted in an immediate blockade of the spontaneous activity of LHRH perikarya during the afternoon of proestrus and significantly decreased LHRH mRNA levels in gonadectomized animals. Taken together, these data suggest that the primary effect of IL-1 beta is at the level of LHRH perikarya, and that the resulting interruption of the cycle is caused by altered LHRH neuronal activity and blunted gonadotropin secretion. PMID- 8243261 TI - The chronic intracerebroventricular infusion of interleukin-1 beta alters the activity of the hypothalamic-pituitary-gonadal axis of cycling rats. II. Induction of pseudopregnant-like corpora lutea. AB - The acute administration of interleukin-1 beta (IL-1 beta) into the brain ventricles of rats has been shown to cause a significant decrease in plasma LH levels, a phenomenon primarily mediated through inhibition of LHRH release. However, there are no studies of the long-term consequences of IL-1 beta injected intracerebroventricularly on the hypothalamic-pituitary-gonadal axis. In particular, we became interested in determining whether IL-1 beta exerts deleterious effects on reproductive parameters, and to what extent they might be caused by a lowering of circulating gonadotropins. In the present experiments, we therefore investigated the effects of the infusion of IL-1 beta to intact cycling female rats and compared them to those observed in rats injected with a potent LHRH antagonist. Although blockade of LHRH receptors caused a modest and delayed inhibition of progesterone secretion, infusion of IL-1 beta (4 ng/h for 4-6 days) was accompanied by persistent and significant increases in plasma P4 levels. In these rats, the PRL release was erratic, with low values during the morning and generally extremely elevated values during the night. The volume of the corpora lutea-I (CL-I) of rats exposed to IL-1 beta, but not to the vehicle or the LHRH antagonist, was significantly increased, and the lutein cells showed extensive hypertrophy. These results indicate that prolonged infusion of IL-1 beta into the brain of cycling rats blocks luteolysis in newly formed CL. These changes were not present in rats injected with the LHRH antagonist, suggesting that they were not primarily related to decreases in gonadotropin secretion. We propose that the high plasma PRL levels may play a role in the changes in ovarian activity which we observed, through other mechanisms, such as sustained increases in adrenal epinephrine and/or glucocorticoids, may also be involved. These findings indicate a novel role for central IL-1 beta in the prevention of luteolysis and the transformation of the CL of the cycle into a CL of pseudopregnancy. PMID- 8243262 TI - Effects of N-terminal, midregion, and C-terminal parathyroid hormone-related peptides on adenosine 3',5'-monophosphate and cytoplasmic free calcium in rat aortic smooth muscle cells and UMR-106 osteoblast-like cells. AB - N-Terminal analogs of PTH-related protein (PTHrP) and PTH bind to a common receptor and exhibit similar biological properties. However, recent studies suggest that certain midregion and C-terminal PTHrP peptides have activities distinct from those of PTH in the placenta and in osteoclasts, respectively. In this study we determined the biological activities of full-length recombinant PTHrP-(1-141) and several synthetic N-terminal, midregion, and C-terminal PTHrP fragments in two PTHrP-producing cell types. Peptides were tested for their ability to stimulate cAMP production and raise intracellular free calcium ([Ca2+]i) in primary rat aortic smooth muscle cells (VSMC) and UMR-106 rat osteoblast-like (UMR) cells. In UMR cells PTHrP-(1-34)NH2, PTHrP-(1-141), and bovine PTH-(1-34) all increased cAMP (approximately 50 fold) and [Ca2+]i (180 nM). By contrast, in VSMC, these N-terminal peptides increased cAMP (3-fold) but had no detectable effect on [Ca2+]i. PTHrP-(1-34) and PTHrP-(1-141) significantly blunted the angiotensin II-induced rise in cAMP (but not the calcium signal) consistent with the concept that PTHrP opposes angiotensin II activity in VSMC. PTHrP-(67-86)NH2, PTHrP-(107-138)NH2, and PTHrP-(107-111)NH2 had no effect on either cAMP or [Ca2+]i in either cell type. VSMC and UMR-106 cells both expressed a 2.5-kilobase PTH/PTHrP receptor messenger RNA (mRNA) transcript. However, high affinity specific binding of 125I-labeled [Tyr36] PTHrP-(1-36)NH2 was detected in UMR cells but not in VSMC. We conclude that the PTH-like, N terminus of the PTHrP molecule is critical in induction of cAMP and [Ca2+]i pathways in UMR cells, and for cAMP stimulation in VSMC. In addition, PTHrP, like other established vasodilators, signals in VSMC mainly (if not exclusively) by increasing the production of cAMP. PMID- 8243263 TI - Growth hormone binding protein in the rat: effects of gonadal steroids. AB - In normal rats, females have higher circulating GH-binding protein (GHBP) levels than males, whereas in the GH-deficient dwarf (Dw) rat, there is no sexual dimorphism in plasma GHBP, suggesting that GH secretion may be involved in this difference. In order to study the relationship between gonadal steroids and GH on GHBP and GH receptor regulation, the levels of plasma GHBP, hepatic bovine GH, and human GH (hGH) binding as well as GHBP and GH receptor messenger RNA (mRNA) have now been studied in normal, Dw, hypophysectomized (Hx), or ovariectomized (Ovx) rats, subjected to different GH and gonadal steroid exposure. In normal male rats, estradiol (E2, 12.5-25 micrograms/day for 1 or 2 weeks) markedly increased plasma GHBP and hepatic hGH, and bGH binding. These effects of E2 were diminished in Dw rats, absent in Hx rats, but restored in Hx rats given exogenous hGH. Plasma GHBP rose in female rats given E2, and fell in females given the anti estrogen tamoxifen. Ovx animals had lower plasma GHBP and hepatic GH binding which was reversed by E2, but not testosterone treatment. Continuous hGH infusions in Ovx rats restored hepatic GH binding, and increased plasma GHBP. In Dw males, hGH increased plasma GHBP and hepatic GH binding, whereas testosterone had no effect on GHBP or GH receptors and did not affect their up-regulation by hGH. Hepatic levels of GHBP-, and GH receptor mRNA transcripts showed the same trends in response to steroid or GH treatment, but the differences were rarely significant, except in Ovx animals which had higher GHBP mRNA transcripts after GH or E2 treatment. Thus E2 and GH increase both plasma GHBP and hepatic GH receptor binding. GH up-regulates GHBP in the absence of E2, whereas E2 treatment does not raise GHBP in the absence of GH. Whereas some of the effects of estrogen could be mediated via alterations in GH secretion, estrogen may also directly influence GHBP production at the liver, but only in the presence of GH. PMID- 8243264 TI - Evidence for a protein-like factor from "rete testis" fluid that suppresses luteinizing hormone-releasing hormone (LHRH) pulses (LHRH statin): a new hormonal activity? AB - Signals that modulate LH-releasing hormone (LHRH) pulse frequency are fundamental mechanisms for regulating important reproductive processes. Gonadal steroids are presently considered to account for the entire gonadal feedback mechanism that modulates LHRH secretion. However, we have previously suggested that a testicular protein(s) present in charcoal-treated rete testis fluid (ctRTF) can suppress LH pulsatility in the ram. The present experiments were aimed at determining whether the disappearance of LH pulses induced by ctRTF administration implicate a hypothalamic or a pituitary site of action. Thus, we have examined the effects of ctRTF peripheral administration on 1) the LH response to LHRH, 2) LHRH portal blood levels, and 3) LHRH content in hypothalamic tissue. Finally, the effects of ctRTF administered into the third ventricle on plasma LH levels were assessed. The present results show that a testicular protein(s) is able to suppress LHRH pulse frequency without affecting amplitude and without any effect on the LH response to LHRH (LHRH Statin). The observation that an active dose administered by the intracerebroventricular route is 0.0005 the active dose needed by the peripheral route reinforces this evidence. These data lead to the new concept that the testicular signals that govern LHRH pulse frequency may be not only steroids, but also proteins. PMID- 8243265 TI - Human recombinant interleukin-1 receptor antagonist prevents adrenocorticotropin, but not interleukin-6 responses to bacterial endotoxin in rats. AB - The present study was designed to analyze the role of endogenous interleukin-1 (IL-1) in the ACTH, corticosterone (CORT), and IL-6 responses of rats to bacterial endotoxin. Recombinant rat IL-1 beta (rIL-1 beta) when given ip resulted in dose-dependent increases in plasma ACTH, CORT, and IL-6 concentrations. Plasma ACTH and CORT responses could be induced by low rIL-1 beta doses that did not elevate plasma IL-6 levels. The half-maximally effective dose of rIL-1 beta was less than 0.6 microgram/kg for the ACTH and CORT responses and higher than 2.5 micrograms/kg for the IL-6 response. Time-course studies indicated that plasma ACTH and CORT concentrations were already elevated 30 min after the injection of rIL-1 beta (2.5 micrograms/kg, ip), with peak values after 1-2 h, followed by a subsequent decline. In contrast, plasma IL-6 concentrations became elevated 2 h after the injection of rIL-1 beta. In another set of experiments, the administration of endotoxin resulted in a dose-dependent elevation of the plasma ACTH, CORT, and IL-6 concentrations. The dose-response characteristics for ACTH, CORT, and IL-6 were different. The half-maximally effective dose for the ACTH and CORT, and IL-6 responses were approximately 2.5 micrograms/kg and more than 10 micrograms/kg, respectively. Time courses of plasma ACTH, CORT, and IL-6 responses to endotoxin (2.5 micrograms/kg, ip) were similar, with peak values measured after 2 h. When given alone, the human IL-1 receptor antagonist (IL-1RA; 1 or 2.5 mg/kg, ip) did not affect resting plasma ACTH and CORT concentrations and reduced plasma IL-6 concentrations in one experiment. At a dose of 1 mg/kg, IL-1RA inhibited ACTH and IL-6 responses to rIL 1 beta (2.5 micrograms/kg, ip) by 75% and 90%, respectively. Administration of IL 1RA (2.5 mg/kg, ip) 30 min after endotoxin (2.5 micrograms/kg, ip) completely prevented the ACTH response and partially inhibited the CORT response, but did not affect the IL-6 response measured 2.5 h after endotoxin administration. We conclude that 1) IL-1 receptors are involved in the ACTH and IL-6 responses to rat IL-1 beta; 2) the ACTH response, but not the IL-6 response, to a low dose dose of endotoxin in rats requires IL-1 receptor activation by endogenous produced IL-1; and 3) circulating IL-6 is not a prime mediator involved in ACTH and CORT responses to low doses of rIL-1 beta and endotoxin. PMID- 8243266 TI - Neither endogenous nor exogenous bradykinin stimulates aldosterone in vivo. AB - It has been suggested that bradykinin (BK) can directly stimulate aldosterone secretion in vitro. Both chronic and acute studies were performed to determine whether we could show a pathway by which BK could stimulate plasma aldosterone in vivo. Chronic studies employed four groups of eight rats fed either a normal sodium or a sodium-restricted diet over 9 days. Half of the rats received infusion of a saline vehicle, and the others received the BK B-2 receptor antagonist HOE-140 at a rate of 200 micrograms/kg.day over 7 days via a sc implanted osmotic minipump. This rate was derived from studies showing that HOE 140 blocked pharmacological doses of exogenous BK. With a normal diet, there was no difference in plasma aldosterone in vehicle-treated rats vs. those treated with HOE-140 (368 +/- 55 vs. 329 +/- 66 pg/ml, respectively). Sodium restriction increased plasma aldosterone 10-fold, but again there was no difference between vehicle-treated rats and those treated with HOE-140 (2964 +/- 439 vs. 3755 +/- 475 pg/ml, respectively). Acute studies employed two groups of six rats. Direct suprarenal intraaortic infusion of 2 micrograms/min BK or saline vehicle was performed over 2 h. Plasma aldosterone was not changed in vehicle-treated groups, but decreased by 22% (P < 0.05) after BK infusion. Thus, blockade of kinin B-2 receptors had no effect on plasma aldosterone in normal or sodium-restricted diet rats. Acute BK infusion did not increase plasma aldosterone. We conclude that although BK may stimulate aldosterone in vitro, we found no physiological correlate to suggest that BK regulates plasma aldosterone in vivo. PMID- 8243267 TI - Cellular mechanism of natriuretic peptides-induced inhibition of endothelin-1 biosynthesis in rat endothelial cells. AB - We studied the cellular mechanism by which natriuretic peptides inhibit the synthesis and release of endothelin-1 (ET-1) in cultured rat aortic endothelial cells (EC). Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) showed dose-dependent and equipotent effects on displacement of [125I]ANP binding and generation of cGMP production in rat EC, whereas C-type natriuretic peptide and biologically inactive ANP analog had lesser effects. ANP and BNP as well as 8 bromo-cGMP had potent inhibitory effects on immunoreactive ET-1 release, the transient increase in the intracellular Ca2+ concentration, and the formation of inositol 1,4,5-trisphosphate stimulated by thrombin in rat EC. A cGMP-dependent protein kinase inhibitor (KT5823), but not a cAMP-dependent protein kinase inhibitor (KT5720), completely abolished the inhibitory effect of ANP on thrombin induced immunoreactive Et-1 release. Northern blot analysis using cDNA for rat prepro-ET-1 as a probe showed that ANP and 8-bromo-cGMP, but not C-type natriuretic peptide, inhibited thrombin-induced prepro-ET-1 mRNA expression, whose effect was abolished by KT5823. These data suggest that ANP and BNP inhibit the thrombin-induced synthesis and release of ET-1 in cultured rat aortic EC by blocking phosphoinositide breakdown, possibly via natriuretic peptides type A receptor-mediated cGMP-dependent mechanism. PMID- 8243268 TI - Evidence that nitric oxide may mediate the ovarian steroid-induced luteinizing hormone surge: involvement of excitatory amino acids. AB - The involvement of excitatory N-methyl-D-aspartate (NMDA) receptors in the hypothalamic control of pituitary LH secretion is well recognized. Recent evidence shows that nitric oxide (NO), a free radical gas, may act as neurotransmitter in the brain, and its efflux is stimulated by activation of NMDA receptors. Studies were undertaken to determine whether NO is involved in the hypothalamic release of LHRH and in the LH surge induced by progesterone (P) in estrogen-primed ovariectomized rats. Rats were ovariectomized and 2 weeks later received estradiol benzoate (30 micrograms sc) at 1000 h. Two days later, P was injected at 1000 h to potentiate the estradiol benzoate-induced LH surge in the afternoon. Serial blood samples were collected at hourly intervals from 1400-1800 h via an intraatrial cannula implanted the day before P injection. Additionally, at various times before onset of the LH surge at 1400 h, the rats were injected sc with one of three inhibitors of NO synthase, the enzyme that generates NO. Control, saline-injected rats showed unambiguous LH surges in the afternoon. However, either a single injection at 1000 h of NG-methyl-L-arginine (20 mg/kg) or three injections at 1000, 1200, and 1400 h of either Nw-nitro-L-arginine methyl ester (NAME, 40 mg/kg) or Nw-nitro-L-arginine (60 mg/kg) to inhibit NO efflux markedly suppressed the P-induced LH surge in the afternoon. To ascertain whether suppression of LH surge was due to blockade of hypothalamic LHRH release, a series of in vitro studies were performed in steroid-primed rats. First we examined the effects of sodium nitroprusside (NPS), a compound that spontaneously generates and releases NO. NPS increased basal and KCl-induced LHRH release in vitro from the medial basal hypothalamus-preoptic area and median eminence fragments. No direct effect of NO at the pituitary level was seen, since NPS did not alter basal or LHRH-induced LH in vitro release from hemipituitaries. In addition, we tested the effects of NAME on NMDA-induced LHRH release in vitro from the median eminence-arcuate nucleus fragments. As expected, NMDA alone (50 mM) induced a significant increase in LHRH release. Addition of NO synthase inhibitor, NAME (1 or 10 mM) to suppress NO efflux, significantly diminished the NMDA-induced LHRH release.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243269 TI - Rana catesbeiana tadpole red blood cells express an alpha, but not a beta, c-erbA gene. AB - Thyroid hormone (TH) receptor number in red blood cells (RBCs) from Rana catesbeiana (RC) tadpoles increases 4-fold during both spontaneous and TH-induced metamorphosis, an effect that we have previously shown to be preceded by an increase in the level of c-erbA-related mRNA. The goals of the present study were to obtain an RC c-erbA alpha cDNA that contains the entire open reading frame for a putative TH receptor protein, to determine if this protein has characteristics typical of a TH receptor, and to assess its contribution to the developmentally related increase in TH receptor number. To accomplish this, the missing 5' sequence of a previously isolated partial RC c-erbA alpha cDNA (RC12) was synthesized by polymerase chain reaction (PCR) and spliced to RC12 to yield a 1490-basepair cDNA (RC15) that contained the entire coding sequence of the receptor protein. Transcription of RC15 followed by translation of its mRNA in a rabbit reticulolysate system yielded a 50-kilodalton protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The protein binds T3 with high affinity (Kd, approximately 0.1 nM), and its affinity for T3 is at least 5 times that for T4. The results of cotransfection studies indicate that RC15 can function as a TH receptor; when COS cells were cotransfected with a construct consisting of RC15 cloned in the expression vector CMV4 and TK28 mult, a construct containing rat GH gene TH response element sequences up-stream of a chloramphenicol acetyltransferase reporter gene, chloramphenicol acetyltransferase activity is expressed in the presence, but not in the absence, of T3. To determine whether RBCs contain any c-erbA beta mRNA transcripts that might contribute to the developmentally related increase in the transcripts detected using RC c-erbA alpha cDNAs, alpha- and beta-specific cDNAs were synthesized by PCR and used as probes in a variety of hybridization assays. In all experiments using conditions in which c-erbA beta transcripts were detectable in other tissues, there was no evidence that tadpole RBCs contained such species. Lack of any beta-specific transcripts was confirmed by PCR, using as template cDNA prepared by reverse transcription of RC RBC RNA. It was also noted that the RBC at metamorphic climax is the tissue with the highest content of alpha specific c-erbA transcripts. It is concluded that the c-erbA alpha gene encodes a TH receptor, and that only the alpha-gene is expressed in tadpole RBCs and subject to regulation during development and by TH. PMID- 8243270 TI - Human thyroid peroxidase-myeloperoxidase chimeric molecules: tools for the study of antigen recognition by thyroid peroxidase autoantibodies. AB - We constructed seven chimeric molecules in which sequential segments in the cDNA for thyroid peroxidase (TPO) were replaced with the homologous regions of myeloperoxidase (MPO) cDNA. The sizes of the translated cDNA segments A through G ranged from 23-175 amino acid residues in length. The TPO-MPO cDNA chimeras, inserted into an eukaryotic expression vector, were stably transfected into Chinese hamster ovary cells. Protein expression was examined by immunoblotting under reduced/denaturing conditions with a murine monoclonal antibody to denatured wild-type TPO. Expression (at a low level) was confirmed for TPO-MPO chimeras A, B, F, and G. The amino acid substitutions in TPO-MPO-C eliminate the monoclonal antibody epitope, and this chimera, therefore, provides a negative control. TPO-MPO-D and TPO-MPO-E did not generate detectable levels of protein. To study TPO autoantibody interaction with native protein, we performed fluorescence-activated cell sorter analysis using intact Chinese hamster ovary cells stably transfected with the wild-type and TPO-MPO chimeric cDNAs. Of the chimeras, only cells transfected with TPO-MPO-A (N-terminal 146 amino acids of MPO substituted for the N-terminal 121 amino acids of TPO) were recognized by TPO autoantibodies, although to a lesser degree than cells expressing wild-type TPO. In conclusion, the present data indicate that TPO autoantibodies can interact with TPO molecules in which the amino-terminus is replaced with the homologous MPO prosequence region, not normally present in mature MPO. Our study provides a foundation for designing future TPO mutants that may be of value for characterizing disease-associated B-cell epitopes in autoimmune thyroid disease. PMID- 8243271 TI - Etidronate inhibits the thyroid hormone-induced bone loss in rats assessed by bone mineral density and messenger ribonucleic acid markers of osteoblast and osteoclast function. AB - TSH-suppressive doses of thyroid hormone are associated with bone loss. We have previously reported that L-T4 decreases femoral, but not vertebral bone mineral density (BMD) in rats. As bisphosphonates are able to decrease bone resorption, especially in high bone turnover states, we investigated the potential effects of etidronate disodium (EHDP) on L-T4-induced bone loss in the rat model by assessing BMD and gene expression of osteoblast (osteocalcin, osteopontin, type I collagen, and alkaline phosphatase), osteoclast (tartrate-resistant acid phosphatase), and cell growth (histone) markers in the skeleton. L-T4 administered for 20 days decreased BMD in the femur, but had no effect on the lumbar spine. EHDP alone had no effect on femoral or vertebral BMD, but did prevent the L-T4-induced bone loss in the femur. L-T4 increased mRNA levels of alkaline phosphatase, tartrate-resistant acid phosphatase, and histone H4 in the femur, but not in the vertebrae. EHDP, which alone had no effect on gene expression in the femur or vertebrae, inhibited the effect of L-T4 on mRNA markers in the femur. The results demonstrate that EHDP can prevent the L-T4 induced decrease in femoral BMD in rats that is associated with the prevention of changes in mRNA markers of osteoclast and osteoblast function. EHDP and other bisphosphonate compounds may be useful in the prevention of thyroid hormone induced bone loss in humans. PMID- 8243272 TI - Human G(olf) alpha: complementary deoxyribonucleic acid structure and expression in pancreatic islets and other tissues outside the olfactory neuroepithelium and central nervous system. AB - G(olf) alpha is a G-protein originally believed to mediate signal transduction exclusively within the olfactory neuroepithelium and subsequently found to be a major stimulatory G-protein in the basal ganglia. Here we present evidence that G(olf) alpha is expressed in several other tissues. The human isoform of G(olf) alpha was isolated from two human insulinoma cDNA libraries. Comparison of the human sequence with rat G(olf) alpha shows 91% nucleotide identity (within the coding region) and 99% identity at the amino acid level. Northern and reverse transcriptase-polymerase chain reaction analyses indicated that G(olf) alpha is expressed in all human insulinomas examined thus far as well as in normal pancreatic islets. G(olf) alpha mRNA was also detected in testis, retina, brain, and liver. Western blot analysis of various mouse tissues demonstrated that the level of G(olf) alpha protein in islets is lower than that in the olfactory neuroepithelium and other parts of the brain; its expression in retina, lung, and spleen was moderately higher than that in islets, and its expression in testis approached that in olfactory neuroepithelium. G(olf) alpha was also detected by immunohistochemistry in mouse islets, human insulinomas, the epithelial lining of mouse epididymis, photoreceptor cells of mouse retina, and mouse lung alveoli. These findings suggest a role for G(olf) alpha in a diverse population of cells located outside the olfactory neuroepithelium and central nervous system. PMID- 8243273 TI - Effects of tumor necrosis factor-alpha on luteinizing hormone-stimulated prorenin production by bovine ovarian thecal cells in vitro. AB - Investigations have been carried out to determine if the cytokine tumor necrosis factor-alpha (TNF alpha), a putative intraovarian regulator, plays a role in the regulation of the ovarian prorenin-renin-angiotensin system. Addition of TNF alpha to cultured bovine thecal cells resulted in a dose-dependent inhibition of LH- or 8-bromo-cAMP-stimulated production of prorenin and renin by the cells in a noncytotoxic manner. No clear inhibitory effect on progesterone production was noted. There was no inhibition of LH- or forskolin-stimulated cAMP formation by TNF alpha. The time-course experiment with TNF alpha revealed that the synthesis, rather than the secretion, of prorenin was inhibited. Also, it was evident that to observe a maximal inhibitory effect, it was necessary to add TNF alpha either before or together with LH. With the increasing delay in the addition of TNF alpha relative to the time of addition of LH, the extent of inhibition gradually decreased, and TNF alpha added 6 h after the addition of LH failed to produce any inhibitory effect. The results obtained permit us to conclude that TNF alpha can counterregulate LH-stimulated prorenin production by thecal cells in culture. The TNF alpha-induced lesion appears to be located at an early step of the biosynthetic pathway of prorenin, which is distal to the activation of LH receptor-coupled adenylate cyclase. Thus, this cytokine appears to be an important intraovarian regulator of prorenin production, a process that is under the stimulatory control of the pituitary gonadotropin. PMID- 8243274 TI - Exposure to physical and psychological stressors elevates plasma interleukin 6: relationship to the activation of hypothalamic-pituitary-adrenal axis. AB - Interleukin 6 (IL-6) is a pleiotropic cytokine produced by the cells of immune and nonimmune origin. Increased production of IL-6 is associated with disturbances of homeostasis, such as trauma, sepsis, or inflammatory diseases. Endotoxemia, tissue injury, or immune inflammatory reactions as well as physical or psychological stress are known to cause increased production of IL-6. We have confirmed this by showing that rats exposed to electric footshock, physical restraint, or a conditioned aversive stimulus have increased levels of plasma IL 6. Interestingly, the kinetics of the increase in plasma IL-6 resembled that of increase in plasma corticosterone. As no detectable endotoxin was found in the plasma samples from stressed and nonstressed rats and there is no evidence of tissue damage and inflammation in situations of restraint or conditioned aversive stimulus, a nonimmune origin of IL-6 is possible. Thus, the releasing of IL-6 into plasma may be under the regulation of neural and endocrine responses to stress. This hypothesis is supported by the decreased production of IL-6 in cultures of splenic cells and peripheral blood mononuclear cells from stressed animals. Furthermore, substantial attenuation of increased plasma IL-6 was achieved by adrenalectomy but not by pretreatment with the beta-receptor antagonist propranolol. The important role of the adrenal gland in the IL-6 response to stress suggests that increased plasma IL-6 may be part of the hormonal responses to stress. As IL-6 induces acute-phase proteins along with glucocorticoids from the adrenal, and regulates the secretion of various hormones from neuroendocrine and endocrine tissues, it is possible that stress-induced increase in plasma IL-6 contributes to the maintenance of homeostasis. PMID- 8243275 TI - Ischemic injury induces brain glucose transporter gene expression. AB - As neurons rely almost exclusively on glucose as an energy substrate, glucose transport is of critical importance to cerebral function. Two specific facilitative glucose transporters, GT1 and -3, predominate in brain, with the latter exclusively expressed by neurons, whereas GT1 is expressed by astrocytes and vascular elements. Little is known about the regulation of these transporters at the genetic level or the extent to which their expression may change in response to acute or chronic changes in metabolic demands. Thus, we employed in situ hybridization to evaluate changes in glucose transporter gene expression in the rat brain in response to ischemia induced by middle cerebral artery occlusion (MCAO). The most remarkable responses were demonstrated by GT1, which within an hour of ischemic insult demonstrated a global increase in gene expression throughout the forebrain. In the ensuing hours, GT1 expression further intensified and became lateralized to the lesioned hemisphere, with normalization of expression contralaterally. Increased GT1 mRNA levels were found in astroglia and microvessels and were also present in distinct neuronal populations, including the piriform cortex, dentate gyrus, and medial habenula, which normally do not express GT1 mRNA. By 24 h post-MCAO, glial cells of the ipsilateral cortex surrounding the infarct zone still demonstrated elevated GT1 mRNA levels, but expression had returned to baseline in neurons. Interestingly, it was not until GT1 expression had subsided (24 h post-MCAO), that there was a modest increase in neuronal GT3 gene expression in the affected hemisphere. GT2 and GT4 mRNAs were not detected in the rat brain under normal conditions or after ischemia. These data demonstrate that ischemia induces an immediate and sustained increase in brain GT1 gene expression in both glial cells and neurons. This augmentation of GT1 expression could represent a defensive strategy aimed at repletion of the brain's energy stores and stabilization of neuronal membrane potential. PMID- 8243276 TI - Characterization and the regulation of inhibin/activin subunit proteins of cultured rat anterior pituitary cells. AB - The production of inhibin/activin by cultured rat anterior pituitary cells was evaluated using specific antisera to inhibin/activin alpha, beta A, and beta B subunit proteins (anti-alpha, anti-beta A, and anti-beta B). Cellular or secreted proteins recognized by the antisera were immunoprecipitated from metabolically labeled cells then analyzed by denaturing polyacrylamide gel electrophoresis. Immunoreactive inhibin/activin beta B proteins were visualized in both cell lysates and the media. Experiments with anti-beta B confirmed that activin-B (beta B beta B) is a local secretory product of cultured rat anterior pituitary cells. The secreted beta B-immunoreactive protein band had an apparent size of 24 25 kilodaltons (kDa) or 14-15 kDa, consistent with the size of unreduced beta B dimer or reduced monomer, respectively. Cell lysates contained two proteins that were specifically immunoprecipitated by anti-beta B. One of these had a mobility of greater than 95 kDa (unreduced) or 55-60 kDa (reduced), probably representing dimers or monomers of the beta B precursor, respectively. The second 14- to 15 kDa (reduced and unreduced) immunoreactive beta B protein band was verified to be the mature beta B monomer. Mature heterodimeric inhibin-B (alpha beta B) was not detected by either anti-alpha or anti-beta B. Multiple protein species, however, were observed to be specifically immunoprecipitated by incubation of cell lysates with anti-alpha. Mature beta A monomer was not detected in any of the samples. The regulation of cellular beta B production was monitored by evaluating its rate of synthesis in pulse-labeled cells. Treatment with either forskolin or 12-O tetradecanoylphorbol acetate enhanced the rate of [35S]cysteine incorporation into the cellular 14- to 15-kDa beta B monomer, indicating that the activation of either protein kinase A or protein kinase C regulates its production. The rate of cellular beta B accumulation was also regulated by activin-A, inhibin-A, and follistatin; activin-A caused a 30% inhibition in contrast to the 70% stimulation by treatment with either inhibin-A or follistatin. Equimolar concentrations of activin-A and follistatin prevented the net effect produced by either factor alone. None of the immunoreactive alpha-forms was detectable under similar pulse labeling conditions, and there was no apparent change in their level after labeling to equilibrium (up to 48 h). The observed changes in beta B accumulation may, therefore, reflect the regulated production of pituitary activin-B. Taken together, these results suggest that locally produced activin-B or gonadal activins exert an inhibitory tone on the production of pituitary activin-B and that this negative-feedback control is in turn modulated by inhibins and follistatins. The relative importance of pituitary and gonadal activins, inhibins, and follistatins in the proposed regulatory loop remains to be established. PMID- 8243277 TI - Activin-A regulates follistatin secretion from cultured rat anterior pituitary cells. AB - The activin-binding protein, follistatin (FS), was immunoprecipitated from metabolically labeled rat anterior pituitary cells or their media using a specific antiserum to purified porcine FS (anti-FS). Several immunoreactive proteins, including one that had a mobility in the range of 42-44 kilodaltons (kDa), were detected in the cell lysates. When immunoprecipitates of the culture medium were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, a broad 35- to 46-kDa or 39- to 53-kDa band was visualized under unreducing or reducing conditions, respectively. Upon deglycosylation by treatment with N-glycosidase-F, the secreted product migrated as a sharp protein band with an apparent size of 35 kDa. The identity or the relatedness of the immunoprecipitated proteins to FS was verified by the ability of the C-terminally truncated form of recombinant human FS (rhFS288) to compete for binding to anti FS. When the cultured rat anterior pituitary cells were treated with either forskolin or 12-O-tetradecanoylphorbol acetate, the accumulation of FS in the culture medium was stimulated by approximately 2.5-fold. These observations suggest that the activation of either the protein kinase A or the protein kinase C signaling pathway has a stimulatory effect on anterior pituitary FS production. A more dramatic stimulation of FS secretion (up to 7-fold) was observed when the rat anterior pituitary cells were treated with activin-A. The concentration dependence for this effect was within the same range that has been reported for most of the actions of activin-A. Inhibin-A suppressed basal FS secretion and blocked its stimulation by activin-A. To determine if locally produced FS exerts an influence on the response of gonadotropes to activins, the effects of anti-FS on FSH secretion were monitored. The ability of this FS antiserum to immunoneutralize the activity of FS was initially confirmed; anti-FS attenuated the inhibitory action of exogenous follistatin on FSH secretion. Treatment of cells with the antiserum increased the apparent sensitivity of gonadotropes to submaximal concentrations of activin-A. Moreover, the presence of the antiserum lowered the concentration of activin-A that was required to produce the maximum amount of FSH secretion, without changing the magnitude of the response. These results suggested that locally produced FS interferes with the secretory response of gonadotropes to activins. Changes in locally secreted FS may, therefore, represent a mechanism by which the response of rat anterior pituitary cells to incoming stimuli are tightly regulated. PMID- 8243278 TI - Tissue distribution of somatostatin receptor subtype messenger ribonucleic acid in the rat. AB - The tissue distribution of mRNA encoding five somatostatin receptor subtypes, SSTR1, SSTR2, SSTR3, SSTR4, and SSTR5, was determined in adult rat tissues by solution hybridization/nuclease protection analysis using sequence-specific cRNA probes. In the central nervous system, SSTR1 and SSTR2 mRNA were expressed widely, with highest levels in hippocampus, hypothalamus, cortex, and amygdala and expression of both isoforms in cerebellum and spinal cord. Expression of SSTR3 was also widespread, occurring in all brain regions examined, with the highest level of expression in the cerebellum. SSTR4 mRNA was detected in most brain regions, with highest levels occurring in the hippocampus, cortex, and olfactory bulb. No detectable levels were found in cerebellum. SSTR5 showed a unique pattern of expression in the central nervous system, being found primarily in the hypothalamus and preoptic area. In peripheral tissues, high levels of SSTR1 and SSTR2 mRNA were found in pituitary and spleen. SSTR1 mRNA was also found in the heart and intestine, SSTR2 was detected in pancreas, and both isoforms were expressed in stomach. Expression of SSTR3 was noted in heart, liver, stomach, intestine, kidney, spleen, and pituitary. The patterns of expression were similar for SSTR4 and SSTR3 mRNA; however, SSTR4 was not expressed in liver. SSTR5 was expressed predominantly in the pituitary, but detectible levels were observed in spleen and intestine. Thus, the SSTR subtype mRNA showed both a tissue-specific and overlapping pattern of expression. Taken together with SSTR-specific signal transduction systems, this probably explains the diverse physiological actions of somatostatin. PMID- 8243279 TI - Expression, regulation, and production of tumor necrosis factor-alpha in mouse testicular interstitial macrophages in vitro. AB - Tumor necrosis factor-alpha (TNF alpha) is a cytokine principally secreted from macrophages and monocytes activated by agents such as lipopolysaccharide (LPS). We have recently shown that TNF alpha inhibited mouse Leydig cell steroidogenesis in vitro. LPS injection has also been shown to repress Leydig cell function and induce TNF alpha messenger RNA (mRNA) expression in testicular interstitial macrophages in vivo. A paracrine regulation of Leydig cell testosterone synthesis by testicular interstitial macrophages via TNF alpha has been proposed. To further support this possibility, we examined whether LPS can induce TNF alpha mRNA expression and protein production in testicular interstitial macrophages in vitro. The regulation of LPS-stimulated TNF alpha mRNA expression in vitro was also investigated by employing the protein synthesis inhibitor cycloheximide (CHX). TNF alpha secretion into culture supernatants was examined by both bioassay and enzyme-linked immunosorbent assay. Isolated testicular interstitial macrophages were cultured for 24 h before the initiation of treatments. Cells were treated with or without LPS (1.0 micrograms/ml) and in the presence or absence of CHX (5.0 micrograms/ml) at different time points. Northern blot analysis showed that TNF alpha mRNA was rapidly and significantly induced by LPS in testicular interstitial macrophages. The peak expression was at 2 h after the treatment, which was 8.3 +/- 2.6-fold over the control (P < 0.05). TNF alpha mRNA then declined quickly and completely disappeared by 8 h after LPS treatment. In contrast to this rapid and transient induction of TNF alpha message by LPS alone, CHX extended the induction and caused a marked increase in LPS-induced TNF alpha mRNA at 2 and 6 h. CHX induced more LPS-stimulated TNF alpha mRNA at 6 h than that at 2 h. At 3 h after LPS treatment, TNF alpha secretion was significantly stimulated (5.6 +/- 1.2 U/micrograms macrophage DNA) measured by L929 tumor fibroblast cytotoxicity. TNF alpha was also detected by enzyme-linked immunosorbent assay in culture media of testicular interstitial macrophages treated with control medium or LPS for 1, 2, and 6 h. TNF alpha secretion was increased in a time-dependent way. There are significantly higher LPS-induced TNF alpha levels in culture media at 2 h (35.4 +/- 2.2 pg/micrograms macrophage DNA) and 6 h (85.5 +/- 11.1 pg/micrograms macrophage DNA) than those in control groups. The current study demonstrates that LPS activates testicular interstitial macrophages to express TNF alpha mRNA and secrete TNF alpha protein in vitro. PMID- 8243280 TI - Bacterial lipopolysaccharide-induced expression of interleukin-6 messenger ribonucleic acid in the rat hypothalamus, pituitary, adrenal gland, and spleen. AB - Whereas the stimulatory effect of interleukin-6 (IL-6) on the hypothalamic pituitary-adrenal (HPA) axis is well established, its mode of action in this axis has yet to be fully elucidated. To further study the role of IL-6 in the HPA axis, we compared the expression of IL-6 messenger RNA (mRNA) in the rat hypothalamus, pituitary, and adrenal gland with that in the spleen after ip or intracerebroventricular (icv) administration of bacterial lipopolysaccharide (LPS). After either ip or icv administration, LPS induced the expression of IL-6 mRNA, which consists of 1.2 kilobases (kb) and 2.4 kb subclasses, in all these tissues of the HPA axis as well as in the spleen. Although we used 100 times less amount of LPS for the icv administration than that used for ip LPS, plasma ACTH levels in both the conditions rapidly reached comparable levels. This icv dose induced IL-6 mRNA expression in the hypothalamus faster than ip dose but also stimulated IL-6 mRNA expression in the hypothalamus, pituitary, and adrenal gland more effectively and smoothly than the ip LPS dose did. Northern blot analysis revealed that in the hypothalamus, pituitary, and adrenals, the predominant subclass of IL-6 mRNA was not 1.2 kb but 2.4 kb. In contrast, this subclass was the minor component in the spleen induced under the same circumstances. These findings indicate that IL-6-synthesizing cells in the HPA axis differ in character from those in the spleen, and that LPS applied in vivo may modulate IL 6 expression in these cells directly and/or indirectly through secondarily activated functions in the neuronal or endocrine systems. PMID- 8243281 TI - Maturation, internalization, and turnover of soluble and membrane proteins associated with atrial myocyte secretory granules. AB - Primary cultures of neonatal atrial myocytes were used to study the biosynthesis of a prominent secretory granule enzyme that occurs naturally in soluble and integral membrane forms. The two most prominent forms of peptidylglycine alpha amidating monooxygenase PAM) in atrial myocytes are type I integral membrane proteins (PAM-1 and -2); smaller amounts of a soluble form, PAM-3, are also found. All three PAM proteins are N-glycosylated, and PAM-1 also has sialylated O linked oligosaccharide. Two hours after their biosynthesis, approximately half of the newly synthesized PAM-1 and PAM-2 proteins have acquired N-linked oligosaccharide chains resistant to digestion with endoglycosidase-H. Secretion of newly synthesized PAM-3 is detectable within 90 min after biosynthesis and is largely complete within 4 h. Release of the catalytic domains of PAM-1 and PAM-2, which requires endoproteolytic cleavage, occurs at a slow rate for many hours after biosynthesis. Release of PAM-3 and the soluble PAM proteins derived from PAM-1 and PAM-2 can be stimulated by secretagogue. Integral membrane PAM proteins that reach the surface of atrial myocytes are internalized and enter the endocytic pathway. The turnover of newly synthesized PAM-1 and PAM-2 is only partially accounted for by the release of soluble PAM protein into the medium and may involve a significant contribution from intracellular degradation. PMID- 8243282 TI - Prolactin receptor messenger ribonucleic acid expression in the ovary during the rat estrous cycle. AB - We have investigated the relative amounts and sites of synthesis during the rat estrous cycle of the two ovarian mRNAs encoding the long and short PRL receptors (PRL-R). Quantitative analysis has revealed that the mRNA encoding the short PRL R is consistently present throughout the cycle in lower quantities than the long receptor mRNA. Both receptor mRNAs are at maximal levels during proestrus, decline to their lowest level of expression during estrus, then gradually rise in metestrus and diestrus. By in situ hybridization, both receptor mRNAs are present during early proestrus in corpora lutea, in the granulosa cell layers of large Graafian follicles, and in the interstitial cells closely associated with these follicles. The short PRL-R mRNA was detected at significant levels in the granulosa-derived cumulus oophorus and in the thecal cell region at this time, whereas the long PRL-R mRNA was only weakly expressed in these cell types. In contrast, the long PRL-R mRNA was present at higher levels, compared to the short receptor mRNA, in the granulosa cells of preantral follicles in the interior of the ovary. On late proestrus, the long PRL-R mRNA was found predominantly in the mural granulosa cells of large Graafian follicles and in corpora lutea, but by estrous morning this mRNA appeared to be mostly restricted to the corpora lutea. This distribution was maintained through estrous evening and metestrous morning. On diestrus, both mRNAs were present in some corpora lutea and in the granulosa cell layer in a subset of the larger Graafian follicles, but were detected at even higher levels in the interstitial cells surrounding these follicles; again, the long receptor mRNA appeared to be only weakly expressed in the thecal cell region of these follicles. These results indicate that the levels and locations of PRL-R mRNA expression in the ovary, and therefore, the potential responsiveness of the ovary to PRL, change throughout the reproductive cycle. Furthermore, the presence of both receptor mRNAs in several different ovarian cell types suggests that both of these receptor forms play important roles in PRL physiology in the ovary. PMID- 8243283 TI - The postnatal serum 3,5,3'-triiodothyronine (T3) surge in the rat is largely independent of extrathyroidal 5'-deiodination of thyroxine to T3. AB - In the rat, selenium deficiency causes a near-complete loss of the selenoenzyme type I 5'-deiodinase (5'D-I), resulting in a marked decrease in hepatic T4 to T3 conversion. In adult rats, serum T4 concentrations are consistently increased, whereas serum T3 and rT3 concentrations are unaffected or slightly decreased and increased, respectively. In rat fetuses near term, serum T4 and rT3 concentrations are not affected by selenium deficiency. We have now studied the effect of selenium deficiency on thyroid function in the neonatal rat. Weanling female rats were fed either a selenium-supplemented or a selenium-deficient diet for 4 weeks before mating and then throughout gestation and lactation. Neonatal rats were killed at 7, 14, 21, and 28 days. Selenium deficiency was confirmed by a more than 89% decrease in liver 5'D-I activity in mothers and pups. Selenium deficiency resulted in significant increases in serum T4 concentrations in 3- and 4-week-old pups. In contrast, selenium deficiency led to a striking increase in serum rT3 concentrations. The normal postnatal serum T3 surge was not affected by selenium deficiency at any age. In 2- and 4-week-old selenium-deficient pups obtained from a second litter from the same mothers, liver 5'D-I activity was markedly decreased, but thyroid 5'D-I activity was not affected. The increased serum rT3 and, less so, T4 concentrations observed in selenium-deficient pups were associated with a significant decrease in brain 5'D-II activity in 14- and 28-day-old pups and in brown adipose tissue 5'D-II activity in 14-day-old pups. In conclusion, the present study demonstrates that the increase in serum T4 concentrations consistently observed in selenium-deficient adult rats occurs only after the second week of life. The normal physiological postnatal 12-fold increase in serum T3 concentrations observed in selenium-deficient pups despite the marked decreases in liver 5'D-I and brain and brown adipose tissue 5'D-II activities suggests that T4 to T3 conversion by peripheral tissues may not be a major source of T3 in the neonate. In contrast, the thyroid gland, whose 5'D-I activity is not affected by selenium deficiency, is probably the principal source of circulating T3 in the neonate. Finally, the early and marked increase in serum rT3 concentrations observed in selenium-deficient pups suggests that liver 5'D-I is important in rT3 deiodination. PMID- 8243284 TI - Estrogen synthesis in vivo in the adult zebra finch: additional evidence that circulating estrogens can originate in brain. AB - Aromatase activity is abundant in limbic and nonlimbic neural structures in zebra finches, especially in the telencephalon near neural circuits that control singing behavior. Also, male songbirds can have high estrogen levels in blood. Because it is difficult to detect aromatase activity in other male tissues, we have postulated that the brain itself is the source of the estrogen present in the blood of males. Previously, we developed methods to measure estrogen synthesis in vivo by injecting [3H]androgen into the systemic circulation or directly into tissues and then determining the quantity of [3H]estrogen entering or leaving the brain. Our results support our hypothesis that the brain is the primary site of estrogen synthesis in males of this species. Here, we confirm that [3H]estrogen emanates from the brain after systemic [3H]androgen injection by showing that the presumptive estrogen in jugular plasma is significantly reduced by treatment with an aromatase inhibitor. In females, but not males, estrogens can be found in large amounts in carotid plasma after systemic androgen injection, presumably due to ovarian estrogen synthesis. However, carotid estrogen levels are variable in females, so it is difficult to determine if the female brain contributes estrogen to blood as is seen in males. The data presented here suggest that the adrenal of males is not an important site of aromatization, because [3H]estrogens are undetected in the adrenals of castrated or sham-castrated males after adrenal injections of [3H] androgen. Nevertheless, at least 2.8- to 17.3-fold more [3H]estrogen was present in the jugular than in the carotid of castrated males after systemic injection of [3H]androgen. Because androgen levels were in the physiological range, we conclude that the brain routinely enriches the estrogen content of blood in normal adult males of this species. PMID- 8243285 TI - In situ demonstration and characterization of progonadotropin-releasing hormone messenger ribonucleic acid in first trimester human placentas. AB - GnRH has been shown to play a role in the regulation of secretion of hCG by human placenta. Immunocytochemical studies have demonstrated the presence of the peptide, but do not address whether the GnRH is maternal, fetal, or placental in origin. In situ hybridization studies using a biotinylated pro-GnRH cDNA were, therefore, undertaken to determine the distribution of pro-GnRH mRNA in first trimester placental samples. Using an avidin-biotin-Cy.5 detection system in conjunction with laser scanning confocal microscopy, pro-GnRH mRNA was shown to be present in both the cytotrophoblast and syncytiotrophoblast cells of placental villi, although not in the cells of the fetal connective tissue. Prior hybridization with a 200-fold excess of unlabeled probe blocked hybridization of the labeled pro-GnRH probe. Southern and sequence analysis demonstrated that the probe hybridized to a transcript identical to hypothalamic GnRH. Immunocytochemical staining using an antiserum to amino acids 6-16 of pro-GnRH demonstrated the presence of translated pro-GnRH in both the cytotrophoblast and syncytiotrophoblast epithelia. We conclude that the synthesis of pro-GnRH by both the cytotrophoblast and the syncytiotrophoblast of human placenta is consistent with either an autocrine or a paracrine mode of GnRH regulation of hCG secretion. PMID- 8243286 TI - Ovarian steroids modulate endothelin-induced luteinizing hormone secretion from cultured rat pituitary cells. AB - It has been demonstrated that endothelins (ETs) induce LH secretory responses in cultured rat pituitary cells. Because estradiol and progesterone are known to be potent modulators of GnRH-induced gonadotropin secretion, we examined whether these steroids also influence the secretory responses of gonadotrophs to ETs. Cultured female rat pituitary cells were treated for 48 h with vehicle (0.2% ethanol), 1 nM estradiol alone, or a combination of 1 nM estradiol and 100 nM progesterone or for 48 h with 1 nM estradiol and a further 4 h with 100 nM progesterone and subsequently stimulated with 10 pM-100 nM ET-1 or ET-3. Forty eight-hour estradiol treatment led to enhanced LH secretory responses to both ETs. This action was facilitated by short term progesterone treatment (3-fold vs. vehicle), while long term progesterone treatment was inhibitory. Perifusion experiments were performed to study the kinetics of individual and pulsatile LH secretory responses after steroid exposure of pituitary cells. ET-1 induced immediate biphasic LH responses that were augmented by long term estradiol treatment. Two-hour progesterone exposure led to marked increases in LH secretion, whereas 48-h progesterone treatment was inhibitory. Estradiol and progesterone were able to modulate both the initial spike and the secondary plateau phase of the secretory profile in response to ET-1, although these actions did not always reach statistical significance. The steroid treatment paradigms employed also induced inhibitory and stimulatory effects in cells that were stimulated with ET-3 in a pulsatile fashion. In these experiments it could be demonstrated that the facilitatory action of progesterone was present after 50 min of treatment and was maximum after 150 min (5-fold enhancement). The present data support the hypothesis that ETs are involved in the physiological regulation of gonadotropin secretion and demonstrate that ovarian steroids can act as potent modulators of ET-induced LH secretion. PMID- 8243287 TI - Differential estrogen substrate specificities for transiently expressed human placental 17 beta-hydroxysteroid dehydrogenase and an endogenous enzyme expressed in cultured COS-m6 cells. AB - The metabolism of estrogens catalyzed by human placental 17 beta-hydroxysteroid dehydrogenase (17HSD) transiently expressed in COS-m6 cells was studied, and the properties of the enzyme were compared with those of an endogenous hydroxysteroid dehydrogenase (HSD) expressed in the cells. In cultured cells, the endogenous HSD had almost exclusively oxidative activity, converting estradiol to estrone (oxidative and reductive activity, 0.84 +/- 0.164 and 0.034 +/- 0.01 nmol/mg protein.h, respectively). This was, nevertheless, opposed to the activity of the transiently expressed human placental 17HSD, as a high reductive activity (0.86 +/- 0.30 nmol/mg protein.h) appeared in the cells after transfection, whereas oxidative activity was not significantly induced. In the different transfections, the reductive activity was induced 13- to 34-fold, and the oxidative activity in the 17HSD-transfected cells was 65-162% of that in the mock-transfected cells. Thus, in cultured cells, these two enzymes preferentially catalyze opposite reactions. When the metabolism of the estrogens was followed up to 20 h, the two enzymes were found to regulate the proportion of estrone to estradiol in the culture medium. The different properties found for the enzymes show that the endogenous HSD expressed in the COS-m6 cells is an additional member of the family of 17HSD enzymes. It is suggested that different 17HSD enzymes exist, with differential estrogen substrate specificities in cultured cells. Thus, in addition to cofactor and substrate availability, the biological activity of estrogens in different cell types may be regulated by the expression of different forms of 17HSD enzymes, resulting in the dominance of either estradiol or estrone production. PMID- 8243288 TI - Prodynorphin gene expression in the rat intermediate pituitary lobe: gender differences and postpartum regulation. AB - The distribution of prodynorphin (proDyn) messenger RNA (mRNA) was examined in the rat pituitary using Northern and in situ hybridization analysis. Anterior pituitary gonadotrophs are known to express ProDyn, but the present study demonstrated that proDyn mRNA was also expressed in the intermediate lobe melanotrophs and was colocalized with POMC mRNA. The 2.6-kilobase proDyn transcript observed in the intermediate lobe was shown to be translatable by polysome analysis. Immunohistochemical studies showed dynorphin (Dyn)-like immunoreactivity in all intermediate lobe melanotrophs. Intermediate lobe proDyn gene expression was not regulated by dopamine, in contrast to intermediate lobe POMC mRNA levels, which were increased with haloperidol and decreased with bromocriptine treatment, as expected. A gender difference in ProDyn gene expression was noted, since intermediate lobes of male rats had nearly 2-fold higher proDyn mRNA levels than intermediate lobes of female rats. In contrast, no gender difference of intermediate lobe POMC mRNA levels were detected. ProDyn mRNA levels were up-regulated by 3- to 4-fold in the intermediate lobes of postpartum females as compared to pregnant or nonpregnant female rats, whereas POMC mRNA levels were unchanged, suggesting a role for intermediate lobe ProDyn in the postpartum period of the female rat. Although our results demonstrate proDyn and POMC coexpression in the pituitary intermediate lobe melanotrophs and show a differential regulational control for each gene in this tissue, the present data also strengthen the notion that proDyn is a precursor that has a role to play in reproductive functions. PMID- 8243289 TI - Differential gene regulation during programmed death (apoptosis) versus proliferation of prostatic glandular cells induced by androgen manipulation. AB - By manipulating the circulating blood level of androgen, it is possible to induce either the programmed death (apoptosis) or proliferation of prostatic glandular cells. To examine the role of differential gene regulation in these two procedure, the expression of the mRNA of a series of genes was quantitated on a per cell basis during the androgen ablation-induced programmed death of rat prostatic glandular cells. These results were then compared to quantitative analysis of the mRNA expression of these same series of genes during the proliferative regrowth of prostatic glandular cells induced in rats castrated for 1 week before being treated with exogenous androgen replacement. These comparisons demonstrated that androgen ablation-induced programmed death of prostatic glandular cells share several (i.e. c-myc, H-ras, and tissue transglutaminase), but not most, of the epigenetic changes associated with androgen-stimulated proliferation of these cells. No enhancement of the mRNA expression of several genes required for entrance of prostatic glandular cells into the S-phase of the proliferative cycle (i.e. histone-H4, c-fos, p53, and ornithine decarboxylase) occurred during androgen ablation-induced programmed death of these cells. These results demonstrated that neither entrance into the S phase nor progression through a defective proliferative cell cycle is involved in androgen ablation-induced programmed death of prostatic glandular cells. This was further supported by the observation that there is a set of genes (i.e. TRPM-2, transforming growth factor-beta 1, alpha-prothymosin, and calmodulin) in which mRNA expression is only enhanced during programmed cell death and not during proliferation of prostatic glandular cells induced by androgen replacement. These results demonstrate that prostatic programmed cell death is a distinct pathway from cell proliferation involving differential gene regulation. PMID- 8243290 TI - Streptozotocin-diabetic rats exhibit facilitated adrenocorticotropin responses to acute stress, but normal sensitivity to feedback by corticosteroids. AB - A variety of chronic stress paradigms have been shown to increase basal activity in the hypothalamic-pituitary-adrenocortical axis, resulting in hypercorticoidism. Despite this, chronically stressed rats typically exhibit facilitated ACTH responses to acute novel stress, suggesting that the activity of some central neural component(s) in the axis is facilitated by chronic stress. We have used the chronic stress of streptozotocin (STZ)-induced diabetes in rats to determine diurnal sensitivity of basal and stimulated ACTH secretion to exogenous corticosterone (B) feedback in vivo. Control and STZ-diabetic rats were adrenalectomized or adrenalectomized and implanted with a 30% or 50% B pellet at the time of vehicle/STZ injection. Rats were killed 5 days later, under basal conditions or after 6 min of restraint, in the morning or evening. We show that basal ACTH secretion in both the morning and evening was similarly suppressed by B in STZ-diabetic and control rats. However, stress-induced ACTH secretion was significantly greater in STZ-diabetic compared to control rats throughout the range 3-7 micrograms/dl B, when tested in the morning. Suppression of evening stress-induced ACTH secretion by B was also significantly different in STZ diabetic rats; however, the IC50 values for the inhibition of ACTH by B did not differ. This result shows that in the evening after stress and under basal conditions in both the morning and evening, sensitivity to B feedback is normal in chronically stressed, STZ-diabetic rats. Despite the observed facilitation of morning stress-induced ACTH secretion in STZ-diabetic rats, there were no differences in hypothalamic CRF content between control and STZ-diabetic tissue. We conclude that 1) the facilitatory input to the paraventricular nucleus functions primarily at the time of the circadian trough to maintain or enhance acute stress responsiveness in chronically stressed, hypercorticoid rats; and 2) the sensitivity of ACTH to inhibition by B is normal in rats chronically stressed by STZ-induced diabetes. PMID- 8243291 TI - Estrogen alters the effects of neuropeptide-Y on luteinizing hormone and follicle stimulating hormone release in female rats at the level of the anterior pituitary gland. AB - In recent years, several studies have shown that neuropeptide-Y (NPY) is involved in the control of LH secretion. We determined the effects of estrogen on NPY induced LH and FSH release in the absence or presence of LH-releasing hormone (LHRH) at the level of the anterior pituitary gland (APG). Adult female rats were ovariectomized. Fifteen to 20 days later, they were given a blank or estrogen filled capsule subdermally and killed 17-19 h later. APG cells were isolated and cultured for 3 days in medium containing 12.5% rat serum collected at death from the same rats used to make the respective APG cell pools. The cells were then challenged for 3 h with vehicle, NPY (10(-12)-10(-6) M), LHRH (10(-9)-10(-6) M), or combinations of NPY (10(-9)-10(-7) M) and LHRH (10(-9) M). LHRH stimulated LH and FSH release from nonestrogen and estrogen-primed cells. NPY at 6.7 x 10(-8) 10(-6) M increased (P < 0.05) LH release and at 10(-6) M increased (P < 0.05) FSH release from estrogen-primed cells, but was without effect on nonestrogen-primed cells. In contrast, NPY at 10(-9)-10(-7) M potentiated the action of LHRH (10(-9) M) to increase the release of LH and FSH from nonestrogen-primed cells, but was without potentiating effects in cultures of estrogen-primed cells. The results demonstrate that 1) NPY can release LH and FSH by a direct action on estrogen primed APG cells; and 2) NPY can potentiate the action of LHRH to increase the release of LH and FSH by a direct action on nonestrogen-primed APG cells. PMID- 8243292 TI - The complementary deoxyribonucleic acid sequence, tissue distribution, and cellular localization of the rat granulin precursor. AB - Granulins (grns; also called epithelins) are cysteine-rich polypeptides with pleiotropic effects on epithelial cell growth in vitro. The grn/epi gene is widely expressed in epithelial cell lines, many of which respond to the gene product, raising the possibility of autocrine or paracrine regulation. In vitro the grn gene is expressed in cell types of diverse lineages, including epithelial cells, lymphoid and myeloid cells, and fibroblasts, but it is not known which cells express the gene in vivo. To understand the physiological role of the grn gene products it is necessary to know the context of grn gene expression in vivo. We have isolated the rat grn precursor complementary DNA and determined, by Northern blot analysis and in situ hybridization, the tissue distribution and cellular localization of grn gene expression. The complementary DNA predicts a 589-amino acid protein of M(r) 63,500 with seven and one-half grn repeats arranged in tandem and shows an overall identity of 75% with human progrn. The grn gene is expressed in a variety of tissues derived from all three embryonic germ layers but is most abundant in the spleen and several tissues of endocrine significance including the adrenal glands, epididymis, placenta, and ovary. Although widely expressed in tissues, gene expression is restricted to specific cell types. For example in the kidney, grn messenger RNA was detected in epithelial cells of the proximal and distal convoluted tubules and Bowman's capsule but not in medullary epithelia. In the spleen, grn messenger RNA expression was localized in lymphocytes, whereas hybridization signals were detected over scattered hepatocytes in the liver. Thus, although the grn gene is widely expressed in tissues and cell lines of many lineages in vitro, its expression in situ is restricted to hematopoietic and some epithelial cells. The restricted cell distribution suggests that the expression of the grn gene is more closely regulated in vivo than in cell cultures. Its localization to epithelial cells in situ supports an autocrine or paracrine role for these factors. PMID- 8243293 TI - Ovarian steroidal response to gonadotropins and beta-adrenergic stimulation is enhanced in polycystic ovary syndrome: role of sympathetic innervation. AB - Experimental induction of a polycystic ovarian syndrome (PCOS) in rodents by the administration of a single dose of estradiol valerate (EV) results in activation of the peripheral sympathetic neurons that innervate the ovary. This activation is evidenced by an increased capacity of ovarian nerve terminals to incorporate and release norepinephrine (NE), an increase in ovarian NE content, and a decrease in ovarian beta-adrenergic receptor number in the ovarian compartments receiving catecholaminergic innervation. The present experiments were undertaken to examine the functional consequences of this enhanced sympathetic outflow to the ovary. The steroidal responses of the gland to beta-adrenergic receptor stimulation and hCG were examined in vitro 60 days after EV administration, i.e. at the time when follicular cysts are well established. EV-treated rats exhibited a remarkable increase in ovarian progesterone and androgen responses to isoproterenol, a beta-adrenergic receptor agonist, with no changes in estradiol responsiveness. Basal estradiol release was, however, 50-fold higher than the highest levels released from normal ovaries at any phase of the estrous cycle. The ovarian progesterone and androgen responses to hCG were enhanced in EV treated rats, as were the responses to a combination of isoproterenol and hCG. Transection of the superior ovarian nerve (SON), which carries most of the catecholaminergic fibers innervating endocrine ovarian cells, dramatically reduced the exaggerated responses of all three steroids to both beta-adrenergic and gonadotropin stimulation. SON transection also reduced the elevated levels of ovarian NE resulting from EV treatment and caused up-regulation of beta adrenoreceptors. Most importantly, SON transection restored estrous cyclicity and ovulatory capacity. The results indicate that the increased output of ovarian steroids in PCOS is at least in part due to an enhanced responsiveness of the gland to both catecholaminergic and gonadotropin stimulation. The ability of SON transection to restore a normal response indicates that the alteration in steroid output results from a deranged activation of selective components of the noradrenergic innervation to the ovary. These findings support the concept that an alteration in the neurogenic control of the ovary contributes to the etiology of PCOS. PMID- 8243294 TI - Gonadal steroid modulation of oxytocin and vasopressin gene expression in the hypothalamus of the osmotically stimulated rat. AB - We investigated the modulatory role of gonadal steroids on the expression of oxytocin (OT) and vasopressin (AVP) cytoplasmic mRNAs in the paraventricular nucleus and supraoptic nucleus of the osmotically stimulated rat. We chronically administered an oral salt load (2% sodium chloride solution for 5 days) to intact and gonadectomized female and male Sprague-Dawley rats and measured serum sodium, body weight, pituitary content of OT and AVP immunoreactivities, and size and abundance of hypothalamic cytoplasmic OT and AVP mRNA transcripts. Intact and gonadectomized rats that were administered an osmotic challenge developed comparable degrees of hypernatremia and loss of body weight as well as depletion of posterior pituitary stores of OT and AVP. Hyperosmolality induced elongation of the OT and AVP transcripts in intact and gonadectomized animals, but only intact rats had enhanced hypothalamic cytoplasmic OT and AVP mRNA concentrations to this stimulus. Replacement with gonadal steroids restored the up-regulation in OT and AVP gene expression in gonadectomized animals rendered hyperosmolar. The findings support a modulatory role for gonadal steroids in hypothalamic OT and AVP gene expression during osmotic stimulation. PMID- 8243295 TI - Differential catabolism of 22-oxacalcitriol and 1,25-dihydroxyvitamin D3 by normal human peripheral monocytes. AB - 22-Oxacalcitriol [1,25-(OH)2-22oxa-D3] mimics the action of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] in a variety of target tissues, including the systemic control of calcitriol metabolism. Similar to 1,25-(OH)2D3, 1,25-(OH)2-22oxa-D3 decreases the rate of 1,25-(OH)2D3 synthesis and accelerates its metabolic clearance rate. We have previously shown that in normal human monocytes, physiological concentrations of 1,25-(OH)2D3 and 1,25-(OH)2-22oxa-D3 determine identical suppression of 1,25-(OH)2D3 synthesis. Moreover, both sterols have a similar potency to induce vitamin D degradation through stimulation of the C24 hydroxylation pathway. In this study, we examined the ability of normal human monocytes to metabolize 1,25-(OH)2-22oxa-D3 and whether the enzymes involved are the same as those that catabolize 1,25-(OH)2D3. Time-course experiments demonstrated no detectable basal catabolic activity. However, exogenous 1,25 (OH)2D3 at physiological concentrations induced 1,25-(OH)2-22oxa-D3 degradation by normal human monocytes. Competition experiments showed that a 10-fold molar excess of unlabeled 1,25-(OH)2D3 inhibited tritiated-1,25-(OH)2-22oxa-D3 catabolism by 85%, whereas a 10-fold excess of unlabeled 1,25-(OH)2-22oxa-D3 reduced tritiated-1,25-(OH)2-22oxa-D3 catabolism by 33%. In contrast, although a 10-fold excess of unlabeled 1,25-(OH)2D3 reduced tritiated 1,25-(OH)2D3 catabolism by 60%, a 1000-fold excess of 1,25-(OH)2-22oxa-D3 was required to reduce tritiated 1,25-(OH)2D3 catabolism to this degree. The apparent Km for 1,25 (OH)2-22oxa-D3 was significantly higher than that of 1,25-(OH)2D3 (2.0 +/- 0.8 0.9 +/- 0.2 nM, respectively; P < 0.001) for the catabolic pathway induced by physiological concentrations of 1,25-(OH)2D3. Moreover, the presence of 0.65 nM 1,25-(OH)2D3 caused an additional increase in the Km for 1,25-(OH)2-22oxa-D3 (3.2 +/- 0.8 nM). These data suggest that 1,25-(OH)2-22oxa-D3 may be less accessible than 1,25-(OH)2D3 to the hydroxylases involved in vitamin D catabolism. The resulting prolonged biological half-life of the analog in certain target tissues may be involved in its selectivity. PMID- 8243296 TI - Effects of calcitriol, 22-oxacalcitriol, and calcipotriol on serum calcium and parathyroid hormone gene expression. AB - Calcitriol markedly decreases PTH gene transcription, but because of the concern about hypercalcemia, there is interest in nonhypercalcemic analogs. We have studied the effects of calcitriol, oxacalcitriol, and calcipotriol on serum calcium and PTH mRNA levels in vivo and in vitro. In vivo in rats, calcitriol was the most effective analog in decreasing PTH mRNA levels, with a maximal effect of about 70% at 25-100 pmol after 24 h. Only 100 pmol led to hypercalcemia. Oxacalcitriol led to a maximal decrease in PTH mRNA levels of 44% at 2 and 5 nmol, similar to 1 nmol calcipotriol. Oxacalcitriol at 200 and 500 pmol led to an increase in serum calcium at 3 h, but not at 6 and 24 h, unlike calcitriol (100 pmol) which caused an increase only at 24 h. In vitro, in primary cultures of bovine parathyroid cells, calcitriol and oxacalcitriol both decreased PTH mRNA levels at similar concentrations. Therefore, in vivo calcitriol is the most effective analog studied in decreasing PTH mRNA levels, including a range of doses that does not cause hypercalcemia. Oxacalcalcitriol and calcipotriol are less effective, but have a wider dose range where they do not cause hypercalcemia. The results in vitro confirm that oxacalcitriol and calcitriol both effectively decrease PTH mRNA levels at the same concentration. The marked activity of calcitriol analogs in vitro compared to in vivo probably reflects differences from calcitriol in their pharmacokinetics. PMID- 8243297 TI - Pituitary adenylate cyclase-activating polypeptide stimulates prolactin gene expression in a rat pituitary cell line. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) has been shown to activate adenylate cyclase and stimulate PRL secretion in dispersed pituitary cells. We have employed the GH3 rat pituitary cell line to investigate whether PACAP can regulate expression of the PRL gene. PACAP increased cellular levels of cAMP in a concentration-dependent fashion (EC50, approximately 6 x 10(-9) M). PACAP also increased PRL mRNA levels in GH3 cells, implying that this peptide stimulates a step in expression of the PRL gene. In addition, PACAP strongly stimulated chloramphenicol acetyltransferase (CAT) activity in GH3 cells transiently transfected with a plasmid containing the first 187 basepairs of the rat PRL promoter cloned up-stream of the CAT gene, implying that PACAP stimulates transcription directed by the PRL promoter. The PACAP stimulation of CAT activity was observed at concentrations as low as 10(-11) M. We examined the action of PACAP on expression of a 5'-deletion series of PRL-CAT constructs. The PACAP response is completely lost when PRL promoter sequences between positions -187 and -113 are removed, implying that neither a previously described sequence resembling a cAMP response element nor the most proximal pit-1-binding site 1P plays a major role in the actions of PACAP on PRL gene transcription. This observation together with the ability of low concentrations of PACAP to stimulate PRL promoter activity without detectably increasing cellular cAMP levels suggest that the action of PACAP on PRL gene transcription might involve a cAMP independent pathway. PMID- 8243298 TI - Characterization of heparin-induced osteopenia in rats. AB - Heparin has been known to induce osteopenia, but its precise mechanism of action is unknown. In the present study, we examined the effect of heparin on the rat femur using single photon absorptiometry and characterized the osteopenia biochemically and pharmacologically. Daily heparin injection dose dependently induced osteopenia in rats. Significant bone loss was observed from 2 weeks after starting heparin treatment (2000 U/kg.day) and peaked at 4 weeks. Serum PTH levels were significantly elevated from 1 week onward after starting heparin treatment, whereas no significant changes were seen in serum total calcium or ionized calcium levels. A bone resorption inhibitor, FR78844 (a bisphosphonate compound), significantly attenuated the heparin-induced osteopenia, as did 1 alpha-hydroxyvitamin D3; with the latter, the effective dose was 10 times lower than that needed for a similar effect against immobilization and ovariectomy induced osteopenia, suggesting an up-regulation of 1 alpha, 25-dihydroxyvitamin D3 receptors in the heparin-treated rats. This speculation was supported by the finding that serum 1 alpha, 25-dihydroxyvitamin D levels were significantly decreased by 54% in the heparin-treated rats compared to those in normal rats. These results suggest that the enhanced bone resorption by high PTH blood levels and the reduction of 1 alpha, 25-dihydroxyvitamin D are involved in the pathogenesis of heparin-induced osteopenia. PMID- 8243299 TI - Effect of ethanol on energy status and intracellular calcium of Sertoli cells: a study using immobilized perfused cells. AB - The effects of ethanol on ATP, O2 consumption, and cytosolic ionized Ca2+ (Ca2+i) were studied in Sertoli cells isolated from the testes of 18- to 21-day-old rats. The cells were immobilized in agarose gel threads and perfused with Dulbecco's Modified Eagle's Medium. Intracellular ATP was determined by 31P nuclear magnetic resonance spectroscopy and enzymatic assay, Cai2+ was measured with the photoprotein aequorin, cell viability was assessed by trypan blue exclusion, and O2 consumption was monitored with a Clark electrode. Ethanol was used with or without pretreatment with the alcohol dehydrogenase inhibitor 4-methylpyrazole (MP). Perfusing the cells for 90 min with 500 mM ethanol produced a 50% reduction in the 31P nuclear magnetic resonance beta ATP signal, and pretreatment with 15 mM MP enhanced this decline of the beta ATP peak to 75%. Enzymatic measurements of ATP revealed that exposure to 500 mM ethanol reduced the ATP levels from 52 +/ 5 to 38 +/- 3 nmol/10(6) cells with MP pretreatment and to 28 +/- 4 nmol/10(6) cells without MP pretreatment (n = 5). Basal O2 consumption was 5.2 +/- 0.5 nmol/min.10(6) cells (n = 5), and it was reduced by ethanol or ethanol plus MP to 4 +/- 0.4 and 3.1 +/- 0.2, respectively (n = 5). The basal concentration of Cai2+ in Sertoli cells was 98 +/- 0.7 nM (n = 32). During perfusion with 500 mM ethanol, Cai2+ increased to 208 +/- 98 nM (n = 5) and was not modified further by the presence of MP. Perfusing the cells for 90 min with 500 mM ethanol with or without MP caused a decrease in cell viability from 93 +/- 2 to 76 +/- 3 and 67 +/- 3, respectively (n = 8). Exposure to 5 mM acetaldehyde produced only a minimal reduction in ATP, with no observable effect at lower concentrations, suggesting that the significant reductions in ATP, O2 consumption, and cell viability evoked by ethanol were not caused by acetaldehyde. These data suggest that ethanol is toxic to Sertoli cells, and its toxicity is not a result of ethanol metabolism. PMID- 8243300 TI - Cholera toxin and dibutyryl cyclic adenosine 3',5'-monophosphate sensitize gonadotropin-releasing hormone-stimulated inositol phosphate production to inhibition in protein kinase-C (PKC)-depleted cells: evidence for cross-talk between a cholera toxin-sensitive G-protein and PKC. AB - The present study assesses the relationship between G-proteins and protein kinase C (PKC) in the gonadotrope. Cells were depleted of PKC with 1 microM phorbol 12 myristate 13-acetate for 12 h, followed by medium 199-BSA for 6 h before treatment with vehicle, pertussis toxin (PTX), cholera toxin (CTX), or (Bu)2cAMP (dBcAMP) for 18 h. PTX (10 ng/ml) significantly decreased GnRH-stimulated inositol phosphate (IP) production over a range of 10(-8)-10(-6) M GnRH. The degree of this inhibition was the same in control cells and PKC-depleted cells. Pretreatment with CTX (0.5 microgram/ml) significantly decreased GnRH-stimulated IP production over a range of 10(-9)-10(-6) M GnRH in PKC-depleted cells. This effect was mimicked by pretreatment with 3 mM dBcAMP. Although CTX and dBcAMP both decreased GnRH-stimulated IP production in control cells, this effect was enhanced in PKC-depleted cells. CTX (0.1 microgram/ml) and dBcAMP (3 mM) both enhanced GnRH-stimulated LH release, whereas PTX (100 ng/ml) had no effect. This was observed in control as well as PKC-depleted cells. Both PKA and PKC are capable of regulating IP turnover by phosphorylating phospholipase-C at distinct sites. CTX activates a G-protein that increases cAMP. cAMP can then activate PKA. In PKC-depleted cells, CTX inhibits GnRH-stimulated IP production. This effect is mimicked by dBcAMP, which suggests a role for PKA in the gonadotrope. The results of this study provide evidence for cross-talk between a CTX-sensitive G-protein and PKC. PMID- 8243301 TI - Identification of binding proteins for nuclear localization signals of the glucocorticoid and thyroid hormone receptors. PMID- 8243302 TI - In vivo measurement of rat ovarian collagenolytic activities. AB - Ovarian collagenases are necessary for the process of ovulation, and they are believed to be activated by the preovulatory LH surge. This information is largely based on in vitro investigations in which the balance between inhibitory and stimulatory principles involved in the activation of collagenase are largely disrupted. Therefore, we developed a simple and reliable method to measure collagenolytic activity in vivo in freely moving rats. By the use of a microdialysis system, a peptide coupled with methyl-coumarin is perfused into the bursa of the ovary. Collagenolytic enzymes cleave this peptide, and the cleaved fragments rediffuse into the microdialysis system. The effluent is collected in fractions, and the peptide-methyl-coumarin complex is cleaved, which results in liberation of fluorescent methyl-coumarin. This assay is linear over a wide range of collagenolytic activity, and other proteases, such as trypsin or plasmin, do not give any fluorescent signal. In proestrous rats, collagenolytic activity increases after the onset of the preovulatory LH surge. In animals in which the LH surge was disrupted by the surgical procedure but had a normal proestrous PRL surge, neither progesterone nor collagenolytic activity increased in the perfusate fluid. This indicates that it is only LH, not PRL, that activates follicular collagenolytic enzymes. Similar results were obtained in immature PMSG/hCG-treated animals. Using a well established zymographic assay, these results were confirmed, and it was further demonstrated that type I and type IV collagenase are active in the rat ovary. PMID- 8243303 TI - Identification and characterization of angiotensin-II receptor subtypes in cultured bovine and human adrenal fasciculata cells and PC12W cells. AB - Angiotensin-II (Ang II) receptor subtypes (AT1 and AT2) were analyzed in bovine adrenal cells (BAC) by binding and cross-linking experiments using [125I]Ang II and [125I]CGP42112, a specific ligand of AT2 receptors. [125I]Ang II binding was reduced by 80% and 20% in the presence of maximal concentrations of the AT1 antagonist losartan (DuP 753) and CGP42112, respectively, whereas [125I]CGP42112 binding was inhibited by Ang II or CGP42112, but not by losartan. In the presence of the reducing agent dithio-1,4-erythritol, the binding of [125I] CGP42112 was increased 2-fold; this was due to an increase in the binding affinity (Kd, 8 +/- 4 x 10(-10) vs. 4.8 +/- 1.2 x 10(-10) M). Cross-linking of [125I]Ang II to BAC in the presence of disuccinimidyl suberate, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, revealed a band of 70,000 +/- 8,000 mol wt (M(r)) under both reducing and nonreducing conditions. This band disappeared when the incubation was performed in the presence of 10(-6) M Ang II or 5 x 10(-8) M CGP42112, but not in the presence of 10(-5) M losartan. Dithio-1,4-erythritol (10 mM) markedly enhanced the band. After cross-linking with 1,5-difluoro-2,4 dinitrobenzene and solubilization of the cells in the presence of protease inhibitors, two radioactive bands were observed with M(r) of 70,000 and 50,000. The first disappeared after the addition of Ang II or CGP42112, whereas the second disappeared in the presence of Ang II or losartan, but not in the presence of CGP42112. Cross-linking of [125I]AngII to either human adrenal fasciculata reticularis cells, which contain only AT1 sites, or COS-7 cells transfected with human AT1 cDNA revealed a major band of 50,000 M(r) that was blunted by Ang II or losartan, but not by CGP42112. Moreover, cross-linking of [125I]Ang II to PC12W cells, which contain only the AT2 receptor subtype, revealed a single radioactive band of 70,000 Mr that was blunted by CGP42112 but not by losartan. Thus, in both BAC and PC12W cells, the AT2 receptor has a M(r) of 70,000, whereas the AT1 receptor in BAC, human adrenal cells, and cells transfected with human AT1 receptor cDNA has a Mr of 50,000. Therefore, the heterogeneity of the size of the Ang II receptor previously reported after photoaffinity or cross-linking was probably due to only to a variation in the degree of glycosylation between tissues and species, but also to the presence of two different receptor subtypes. PMID- 8243304 TI - Reproductive aging in the male brown-Norway rat: a model for the human. AB - Previous studies in the Sprague-Dawley rat have demonstrated that male reproductive aging is primarily a neuroendocrine dysfunction characterized by decreased pulsatile LH secretion leading to low serum testosterone (T) levels, whereas sperm production is relatively well maintained. In contrast to the Sprague-Dawley rat, the Brown-Norway (BN) rat has a longer life span, does not become obese, and experiences fewer age-related tumors of the endocrine or reproductive system, thus providing a disease-free model for studying male reproductive aging. We studied the changes in serum hormone levels and related these alterations to testicular T production, Leydig cell morphometry, and spermatogenesis in young (6 months), aging (22 months), and old (30 months) BN rats. Low serum T levels were associated with decreased LH levels in the 22-month old (T, 0.58 +/- 0.08 ng/ml; LH, 0.45 +/- 0.06 ng/ml) and 30-month-old rats (T, 0.63 +/- 0.10 ng/ml; LH, 0.34 +/- 0.04 ng/ml) compared to those in young rats (T, 1.35 +/- 0.30 ng/ml; LH, 0.79 +/- 0.10 ng/ml). In vitro Leydig cell T production, basally and after stimulation by LH, was similar in young and old rats. The total testicular T content was lower in 30- than in 6-month-old rats. Testicular morphometry showed smaller Leydig cell volume in the old (857 +/- 97 microns3) than in the young rats (1387 +/- 103 microns3), although their number per testis remained unchanged (6 months, 22.7 +/- 1.6 x 10(6)/testis; 30 months, 25.2 +/- 3.1 x 10(6)/testis). In contrast, a marked (68.4%) reduction in the total number of Sertoli cell per testis was noted in the 30-month-old rats. The proportion of the testis occupied by seminiferous tubules was also reduced in the old rats. Significant (P < 0.05) age-related reductions occurred in tubule diameter, length of tubules, and volume of tubules and their lumens. The testicular sperm concentration and total sperm production were significantly reduced in the 22- and 30-month-old rats. These changes in seminiferous tubule function in the old rats were associated with low serum and testicular inhibin and high serum FSH levels. We conclude that aging in the reproductive axis of the BN rat is manifested by 1) lower serum T levels due to decreased pituitary LH stimulation of endogenous T production, and 2) decreased seminiferous tubule function accompanied by elevated FSH levels indicative of a primary testicular disorder.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243305 TI - Impaired growth hormone-releasing hormone signal transduction in the dwarf (dw) rat is independent of a generalized defect in the stimulatory G-protein, Gs alpha. AB - The homozygous dwarf (dw) rat pituitary is characterized by a 95% reduction in GH content as well as a 75-80% reduction in the number of somatotrophs. The nature of the mutation responsible for this phenotype is unknown. Previous investigations from our laboratory indicate that dw somatotrophs exhibit decreased sensitivity and a reduced GH secretory response to GH-releasing hormone in vitro, accompanied by a decreased generation of cAMP. We hypothesized that dw rats have a defect in the pathway linking the GH-releasing hormone receptor to adenylyl cyclase and focused on the expression and function of the stimulatory G protein, Gs alpha. When corrected for differences in pituitary size and cell number, GH mRNA content was reduced by 78% compared to that in controls. However, there was no difference in Gs alpha mRNA content or size in dw pituitaries. Similarly, there was no difference in the content or size of mRNA for the pituitary transcription factor pit-1 in dw pituitaries. Immunoblot analysis of pituitary membrane proteins using a Gs alpha-specific antibody revealed no differences in size, quantity, or relative distribution of Gs alpha peptides between control and dw pituitaries. In addition, cholera toxin effectively ribosylated dw Gs alpha, and there were no differences in the size, quantity, or relative distribution of ribosylated membrane proteins between dw and control pituitaries. Finally, to examine for mutations in other regions of the Gs alpha coding sequence, we cloned the full-length Gs alpha cDNA from dw rat pituitaries by polymerase chain reaction. The sequence of this clone was identical to that of normal rat Gs alpha cDNA. These results indicate that 1) 52-kilodalton Gs alpha appears to predominate in both normal and dw pituitary; 2) the content, function, and sequence of Gs alpha in adult dw rats are normal; and 3) a generalized Gs alpha regulatory or structural mutation as the cause of the observed phenotype can be excluded. The results also demonstrate that there is no decrease in pit-1 expression in the adult dw pituitary. PMID- 8243306 TI - The estrogen antagonist ICI 182,780 reduces cancellous bone volume in female rats. AB - Although estrogen is thought to protect the skeleton by inhibiting bone resorption, we have also found that in the rat, estrogen stimulates cancellous bone formation. However, the extent to which the various skeletal actions of estrogen are mediated by classical estrogen receptors remains unclear. Although estrogen receptor antagonists such as tamoxifen have been used to study this question, interpretation of the results is complicated by the fact that this agent also acts as a partial estrogen agonist. However, the recent development of estrogen antagonists devoid of agonist activity provides an opportunity to explore this question further. We, therefore, investigated the effect of administration of the pure estrogen antagonist ICI 182,780 on the skeleton of adult female rats. We found that ICI 182,780 reduced bone volume at the proximal tibial metaphysis by approximately 30%, associated with an increase in osteoclast surface. We then investigated the effect of ICI 182,780 on the anabolic action of estrogen. We used ovariectomized rats treated with 3-amino-1-hydroxypropylidene-1 bisphosphonate to inhibit bone resorption, thereby preventing any increase in bone formation as a result of the stimulation of bone resorption due to estrogen deficiency. 17 beta-Estradiol (1 micrograms/kg) stimulated cancellous bone formation in such animals by approximately 8-fold; this increase was abolished when ICI 182,780 was also given. In contrast, ICI 182,780 affected neither longitudinal nor periosteal tibial growth in either intact animals or ovariectomized rats given estradiol or vehicle. We conclude that ICI 182,780 reduces cancellous bone volume in the rat by antagonizing estrogen's actions on bone formation and resorption, suggesting that these processes are both mediated by classical estrogen receptors. PMID- 8243307 TI - Enhancement of growth hormone bioactivity by zinc in the eluted stain assay system. AB - The effects of ionic zinc (Zn2+) on human (h) GH bioactivity have been examined using a lactogenic bioassay. The potencies of pituitary-derived hGH (IRP 80/505), recombinant 22K hGH (IRP 88/624), pituitary-derived human PRL (IRP 84/500), and a recombinant methionyl 20-kilodalton variant of hGH in the presence of selected concentrations of ZnCl2 were investigated with an eluted stain assay that uses Nb2 rat lymphoma cells. This precise colorimetric bioassay is based upon the reduction of a yellow tetrazolium salt, 3-[4,5-dimethyl-thiazol-2-yl]2,5-di phenyl-tetrazolium bromide, to its purple formazan by lactogen-activated Nb2 cells. Zinc (6-100 microM) enhanced the bioactivity of low doses (< 0.045 nM) of both pituitary-derived and recombinant 22K hGH, although the magnitude of enhancement was considerably less than might have been anticipated from previous binding studies (13). Higher concentrations of pituitary-derived hGH (> 0.045 nM) were inhibited by Zn2+. The bioactivity of recombinant methionyl 20K hGH was greatly enhanced by zinc (3-100 microM). In contrast to hGH, the bioactivity of hPRL was not potentiated by Zn2+. These discriminatory effects of Zn2+ when stimulating via the lactogenic receptor are in concordance with the results of previous radioligand binding studies (13). The striking enhancement of 20K hGH lactogenic bioactivity was observed at Zn2+ concentrations within the physiological range for normal human serum (5-20 microM). PMID- 8243308 TI - The growth hormone (GH)-binding protein cloned from human IM-9 lymphocytes modulates the down-regulation of GH receptors by 22- and 20-kilodalton human GH in IM-9 lymphocytes and the biological effects of the hormone in Nb2 lymphoma cells. AB - The cDNA coding for the 246-amino acid long N-terminal extracellular portion of the human (h) GH receptor, corresponding to the circulating GH-binding protein (hGHBP), was cloned by polymerase chain reaction from human IM-9 lymphocytes. The cDNA sequence was identical to that reported for human liver and placenta and demonstrated alternative splicing of exon 3. The protein with the exon 3-encoded domain was expressed and secreted in glycosylated form from baby hamster kidney (BHK) cells, purified to homogeneity, and sequenced; the amino acid sequence was identical to that predicted from liver cDNA. The cloned hGHBP competed in a dose dependent fashion for binding of 125I-labeled 22-kilodalton (kDa) hGH, and at higher concentrations for binding of 125I-labeled 20-kDa hGH, to IM-9 lymphocytes. hGHBP decreased the association rate of [125I]hGH to the cells without decreasing the dissociation rate. hGHBP blocked the down-regulation of GH receptor in IM-9 cells by both 22- and 20-kDa hGH. hGHBP also blocked the binding of [125I]hGH to PRL receptors on Nb2 lymphoma cells and the effect of the hormone on thymidine incorporation. Binding of both 22- and 20-kDa hGH to the binding protein was demonstrated directly by immunoprecipitation with monoclonal antibody 263. The present work thus establishes the identity of the IM-9 human GHBP from those of liver and placenta, and demonstrates its ability to bind both 22- and 20 kDa hGH with good affinity and to block their biological actions mediated though both somatogenic and lactogenic receptors. The modulation of receptor down regulation by the BP may be a relevant facet of its physiological role. PMID- 8243309 TI - Estrogen uncouples beta-adrenergic receptor from the stimulatory guanine nucleotide-binding protein in female rat hypothalamus. AB - The responsiveness of adenylyl cyclase to beta-adrenergic receptor stimulation was investigated in membranes prepared from hypothalamus-preoptic area and cortex of ovariectomized female rats injected with oil vehicle or estradiol benzoate 24 or 48 h before death. Membranes from the hypothalamus-preoptic area of ovariectomized animals displayed a concentration-dependent stimulation of adenylyl cyclase when incubated with the beta-adrenergic receptor agonist, isoproterenol (10(-7)-10(-5) M). This response was suppressed in membranes from estrogen-treated animals. The effect of estrogen was observed 48 h, but not 24 h, after hormone administration. In addition, estrogen had no measurable effect on hypothalamic adenylyl cyclase activation by either GTP (10(-8)-10(-5) M) or forskolin (10(-8)-10(-6) M), on beta-adrenergic receptor density, or on antagonist binding affinity measured with the beta-adrenergic antagonist [125I]iodocyanopindolol. Analysis of isoproterenol displacement of iodocyanopindolol binding revealed that estrogen reduced agonist binding affinity in hypothalamus-preoptic area membranes. In membranes from ovariectomized controls, high affinity agonist binding to the beta-adrenergic receptor was apparent and was abolished by guanine nucleotides. However, membranes from estradiol-treated rats demonstrated only low affinity agonist binding that was unaffected by guanine nucleotides. Estradiol did not detectably alter concentrations of either cholera or pertussis toxin substrates in hypothalamus preoptic area membranes. These data indicate that estrogen promotes a stable time dependent desensitization of beta-adrenergic receptor activation of adenylyl cyclase in hypothalamus and preoptic area by uncoupling the receptor from the guanine nucleotide-binding protein, G8. PMID- 8243310 TI - The effects of hydrogen peroxide on steroidogenesis in mouse Leydig tumor cells. AB - It has previously been reported that treatment of rat luteal cells and human granulosa luteal cells with hydrogen peroxide (H2O2) results in a significant inhibition of steroid production. The mechanism of inhibition in the former was found to be at the level of cholesterol transport into the mitochondria, whereas in the latter it was found to be a result of inhibition of one or more enzymes in the steroidogenic pathway. In the present study we examined the effects of H2O2 on hormone-stimulated steroid production in another steroidogenic cell type, the Leydig cell. Our results demonstrate that treatment of either LH- or cAMP analog [(Bu)2cAMP]-stimulated MA-10 mouse Leydig tumor cells with H2O2 results in a dose dependent inhibition of the production of progesterone (the major steroid produced in MA-10 cells). It was also observed that similar concentrations of H2O2 resulted in a significant inhibition of protein synthesis, a finding which could in part be responsible for the observed decrease in steroid production. Furthermore, although H2O2 resulted in a dose-dependent inhibition of (Bu)2cAMP stimulated pregnenolone production, addition of the hydroxylated intermediate 22R hydroxycholesterol and inhibitors of further pregnenolone metabolism demonstrated that cholesterol side-chain cleavage complex activity was unaffected by H2O2. Conversely, incubation of H2O2-treated cells in the presence of pregnenolone resulted in a very significant inhibition of progesterone synthesis. These data indicate that H2O2 inhibits steroidogenesis in Leydig tumor cells primarily by inhibiting the activity of the 3 beta-hydroxysteroid dehydrogenase, but that other effects of H2O2 such as inhibition of protein synthesis and the transfer of cholesterol to the cholesterol side-chain cleavage complex may also be involved. PMID- 8243311 TI - Evidence for the potentiation of epidermal growth factor receptor tyrosine kinase activity by association with the detergent-insoluble cellular cytoskeleton: analysis of intact and carboxy-terminally truncated receptors. AB - The ligand-stimulated tyrosine kinase activity of the normal human epidermal growth factor (EGF) receptor and a truncated EGF receptor lacking 164 carboxy terminal (C-terminal) amino acids was examined in intact cells and after Triton X 100 extraction into Triton-soluble and -insoluble (cytoskeletal) preparations. Detergent extraction of the intact and truncated receptors appeared complete using 0.3% Triton as demonstrated by anti-EGF receptor immunoblots, tyrosine kinase assays, and marker enzyme (alkaline phosphatase) solubilization. Higher Triton concentrations yielded no additional EGF receptor extraction and began to inhibit EGF-stimulated kinase activity toward angiotensin II (AII). Furthermore, the tyrosine kinase activity of the truncated EGF receptor exhibited increased sensitivity to Triton extraction, suggesting a lower affinity or a more labile association of this receptor with the cytoskeleton. However, both EGF receptor forms had altered catalytic activity when associated with the cytoskeletal fraction, as evidenced by the increased phosphorylation of the exogenous substrates: AII, src-peptide, and [Val5]AII. Kinetic analyses of both receptor types revealed that the cytoskeletal fractions obtained using 0.3% Triton contain EGF receptor activity that exhibits a Michaelis-Menten constant (Km) for AII that is 2- to 3-fold more favorable than that calculated for the soluble receptor forms. EGF treatment of intact cells containing either the intact or truncated receptor revealed similar phosphorylated proteins in the soluble fraction of both cell types, although there was evidence for the enhanced phosphorylation of certain proteins (e.g. 115 and 50 kilodalton proteins) in cells containing the truncated receptor. There was also a greater number of tyrosine-phosphorylated proteins in the Triton-insoluble fraction of cells containing the truncated receptor, suggesting an altered specificity of this receptor toward selected cytoskeletal proteins. This work indicates that EGF receptor-cytoskeletal interaction may be an important consideration in the control of receptor-kinase activity and has examined the detergent sensitivity of this association. These studies also suggest that the C-terminal domain of the EGF receptor may affect cytoskeletal interaction in addition to influencing the receptor's catalytic capacity. PMID- 8243312 TI - Gastric inhibitory polypeptide receptor, a member of the secretin-vasoactive intestinal peptide receptor family, is widely distributed in peripheral organs and the brain. AB - Gastric inhibitory polypeptide (GIP), or glucose-dependent insulinotropic peptide, is released from endocrine cells in the small intestine after meals. It is involved in several facets of the anabolic response and is thought to be particularly important in stimulating insulin secretion. We have cloned, functionally expressed, and mapped the distribution of the receptor for GIP. It is a member of the secretin-vasoactive intestinal polypeptide family of G-protein coupled receptors. When expressed in tissue culture cells, it stimulates cAMP production (EC50 0.3 nM) and also increases intracellular calcium accumulation. GIP receptor mRNA is present in the pancreas as well as the gut, adipose tissue, heart, pituitary, and inner layers of the adrenal cortex, whereas it is not found in kidney, spleen, or liver. It is also expressed in several brain regions, including the cerebral cortex, hippocampus, and olfactory bulb. These results suggest that GIP may have previously undescribed actions. GIP receptor localization in the adrenal cortex suggests that it may have effects on glucocorticoid metabolism. Neither GIP nor its effects have been described in the central nervous system, and mRNA for the known peptide ligand for the receptor cannot be detected in the brain by in situ hybridization or polymerase chain reaction. This suggests that a novel peptide may be present in the brain. PMID- 8243313 TI - Effects of oleate and fatty acids from omental adipocytes on insulin uptake in rat liver cells. AB - Hyperinsulinemia in central type (upper body) obesity has been related to reduced hepatic insulin uptake caused by fatty acids reaching the portal vein directly from the omental (visceral) fat depot. We investigated the effect of sodium oleate and fatty acid-rich buffer recovered from incubation of omental adipocytes with 10(-7) M epinephrine on specific [125I]insulin uptake (at 37 C) in freshly prepared and cultured hepatocytes and nonparenchymal liver cells isolated from 200-250 g rats. In hepatocytes, insulin uptake was studied after standard cell isolation and washing and after additional repeated 30-min washings at 37 C. In the vigorously washed hepatocytes, both fatty acid rich buffer (from 0.08-1.3 mM) and sodium oleate (from 0.0125-1.2 mM) decreased insulin uptake maximally by approximately 30% and approximately 40%, respectively (P < 0.005); a biphasic effect of fatty acids was observed, with maximal inhibition occurring at 0.08-0.1 mM. Without vigorous washing, oleate had no effect on insulin uptake in either freshly prepared or cultured hepatocytes and nonparenchymal cells. The reason for the observed inhibitory effect of fatty acids on insulin uptake in vigorously washed cells remains unknown, but the effect is unlikely to be physiologically important. Thus, our data do not support the hypothesis that the increased concentration of fatty acids in the portal vein is responsible for reduced hepatic insulin uptake and hyperinsulinemia in upper body obesity. PMID- 8243314 TI - Induction of ovarian follicle luteinization by recombinant follicle-stimulating hormone. AB - Ovulation and subsequent luteal tissue formation are preceded by midcycle surges of both LH and FSH. Although LH has been widely known as the luteinizing hormone, a potential role for FSH in the luteinization process is possible. Our earlier studies using recombinant FSH (rcFSH) without LH contamination have shown that treatment with a surge dose of rcFSH induces ovulation of mature follicles in hypophysectomized rats. The present studies examined further whether FSH alone is sufficient to induce normal corpus luteum formation. Immature hypophysectomized rats were implanted with an estrogen pellet (10 mg diethylstilbestrol). Two days later, a minipump releasing 4 IU rcFSH/day was placed to induce follicular growth. Forty-eight hours after FSH treatment, both DES pellet and FSH minipump were removed, and rats were injected with a single sc dose of 40 IU rcFSH, 5 micrograms hCG, or saline. For some animals, oviducts were excised the following day to determine the number of ovulated oocytes. The remaining animals received, 2 days later, sc injections of 125 micrograms ovine PRL twice daily for 3 days to maintain luteal function. All rats that received a surge dose of rcFSH or hCG ovulated similar numbers of oocytes, whereas none of the control animals did. Ovaries and blood samples were obtained 5 days after the gonadotropin surge. rcFSH and hCG significantly increased ovarian weight to 73.9 +/- 4.8 and 94.7 +/- 5.6 mg, respectively, compared to 10.0 +/- 0.5 mg in controls. Serum progesterone levels were increased by 192- and 102-fold in rcFSH- and hCG-treated animals, respectively, compared with those in the saline-treated rats. rcFSH and hCG also induced a marked elevation of ovarian [125I]hCG binding (4.2 +/- 0.2 and 3.7 +/- 0.1 ng/mg ovary, respectively), whereas ovaries from control animals exhibited low binding (0.6 +/- 0.1 ng/mg ovary). These gonadotropin-induced increases in [125I]hCG binding were associated with similar elevations in the levels of three LH receptor transcripts of 2.5, 4.2, and 7.0 kilobases. Also, levels of the ovarian cholesterol side-chain cleavage enzyme (CYP 11A) mRNA (2 kilobases) were low in control animals, but increased 20.5- and 14.3-fold after surge doses of rcFSH and hCG, respectively. Accompanied by biochemical signs of luteinization, morphological features typical of luteinized ovaries were found in both rcFSH and hCG groups, showing the formation of large polyhedral lutein cells and small spindle-shaped lutein cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243315 TI - Evidence that the inhibitory effects of iodide on thyroid cell proliferation are due to arrest of the cell cycle at G0G1 and G2M phases. AB - The thyroid gland is unique in its ability to respond to ambient levels of iodine to autoregulate thyroid function and, possibly, thyroid cell proliferation. Although the inhibitory effects of iodide on thyroid cell proliferation have been previously reported, the exact mechanism and site of action of iodide on cellular proliferation events are poorly understood. Our initial experiments established the optimal cell plating density and timing to achieve exponential cell growth of FRTL5 thyroid cells, and subsequent studies using flow cytometric DNA analysis established the normal cell cycle kinetics of FRTL5 thyroid cell proliferation. FRTL5 cells were then exposed to graded concentrations of sodium iodide to establish whether the inhibitory effects of iodide are mediated through specific cell cycle events. We observed that increasing concentrations of iodide inhibited FRTL5 thyroid cell proliferation. Analysis of the cell cycle revealed two specific effects of iodide on cell cycle kinetics. The first was an arrest of cells in G0G1, evidenced by an accumulation of cells in this phase and a concomitant reduction in the percentage of cells in the S-phase. The second was an arrest of cells in the G2M phase of the cycle. G0G1 and G2M arrest occurred within 24 h and then reached a plateau. Iodide exposure did not increase the number of cells undergoing necrosis. The addition of methimazole at two concentrations (0.2 and 2 mM) to cells exposed to 100 mM NaI prevented the accumulation of cells in G2M, but did not abolish the accumulation of cells in G0G1 or the reduction in cell number. These results indicate that the inhibitory effects of iodide on FRTL5 thyroid cell proliferation are mediated by its action at two critical regulating points of the cell cycle, G0G1 and G2M. It appears that organified iodine may mediate the cell cycle arrest in the G2M phase, whereas inorganic iodide may be responsible for the inhibitory effects at G0G1. PMID- 8243316 TI - Coexpression of atrial natriuretic factor and beta-endorphin in a subpopulation of rat splenic macrophages: age-related differences. AB - Separate demonstration of immunoreactive (ir) atrial natriuretic factor (ANF) and ir-beta-endorphin (beta EP) in macrophages of the rat spleen prompted a reexamination of their distribution and cellular localization in addition to their developmental regulation. Double labeling immunohistochemistry was carried out on paraffin-embedded spleen sections of adult male Sprague-Dawley rats. Cells stained positive with antiserum (S118) raised against rat ANF-(99-126) were colocalized in more than 95% of cases with immunofluorescent staining of ir beta EP-(1-31) and distributed sparsely throughout the venous sinusoidal regions of the red pulp. In 1- or 5-day monolayer cultures of adherent splenocytes, 11-16% of the cells stained positive for either irANF or ir beta EP. Under these conditions, more than 95% of irANF- and ir beta EP-positive cells were also fluorescence stained for the histiocyte marker S22 or the rat macrophage marker ED-1. Colorimetric in situ hybridization similarly revealed signals for pro-ANF mRNA in more than 95% of ir beta EP-positive adherent cells. Thus, taken together with previous reports, our present findings suggest that in the rat spleen, both ANF and beta EP are produced by the same population of macrophages. However, the tissue levels and processing of the two peptides over the developmental period of the animal differed markedly in several ways. In splenic extracts of 2-day-old neonatal rats, Northern blot analysis and RIA revealed a greater abundance of pro ANF mRNA signal and an irANF concentration about 6-fold greater than those in 30 day-old animals. In contrast, the ir beta EP concentration did not vary significantly over the same period, consistent with the level of POMC mRNA. Sephadex G-50 gel chromatography of splenic extracts from 2-day-old animals revealed predominantly higher mol wt forms of irANF and ir beta EP. From days 16 60, a significant proportion of irANF eluted in the same fractions as the mature circulating form, rat ANF-(99-126); the proportion of 3.5-kilodalton ir beta EP increased progressively to become the predominant species in adult tissues. Thus, it appears that although ANF and beta EP are coexpressed by the same population of splenic macrophages, they differ markedly during the developmental period with respect to their constitutive regulation. PMID- 8243317 TI - Embryonic stem cells express growth hormone receptors: regulation by retinoic acid. AB - Embryonic and fetal growth are generally considered to be independent of pituitary GH. However, it has been demonstrated recently that 18-day-old rat embryos and rat fetuses express GH receptors, suggesting that GH could play a role in early development. The aim of the present investigation was to determine whether preimplantation embryos also express GH receptors. Germ line competent mouse embryonic stem (ES) cells and cultured mouse preimplantation embryos were examined with Northern blot analysis, RNAse-protection solution-hybridization assays, reverse transcription/polymerase chain reaction assays and immunohistochemistry for the detection of GH receptors. Northern blot analysis of ES cells using a probe corresponding to the extracellular domain of the GH receptor demonstrated the presence of two transcripts (1.2 and 4.5 kilobases). The RNAse-protection solution-hybridization assay revealed that ES cells express approximately one sixth of the GH-receptor messenger RNA (mRNA) levels expressed in liver from pregnant mice. Treatment of cultured ES cells with retinoic acid (100 nM) for 6 days increased GH-receptor mRNA levels (P < 0.01). GH-receptor mRNA was further identified in ES cells, preimplantation embryos, muscle, liver, and placenta by a reverse transcription/polymerase chain reaction assay. In humans it has previously been shown that exon 3 of the GH-receptor is deleted in the placenta. However, none of the studied mouse tissues had a deletion of the GH receptor mRNA corresponding to exon 3 of the human GH receptor. GH-receptor immunoreactivity was identified on the cultured ES-cells by immunohistochemistry. In conclusion, we have in the present study shown that germline competent ES cells and preimplantation mouse embryos express the GH receptor transcript and that this transcription is increased by retinoic acid in ES cells. Furthermore, the presence of GH-receptor immunoreactivity on the ES cells indicates that the GH-receptor transcript is translated. PMID- 8243318 TI - Evidence that changes in the function of the subtypes of the receptors for gamma amino butyric acid may be involved in the seasonal changes in the negative feedback effects of estrogen on gonadotropin-releasing hormone secretion and plasma luteinizing hormone levels in the ewe. AB - We have investigated the effects of gamma-amino butyric acid (GABA) agonists and antagonists to GABAA and GABAB receptors microinjected (1 microliter) into the medial preoptic area (MPOA) on LH secretion in ovariectomized (OVX) ewes with or without estrogen (E) treatment, during the nonbreeding season. Guide tubes (19 gauge) were placed into the MPOA of OVX ewes and injections of 1 microgram or 10 micrograms of the GABAA agonist muscimol or the GABAA antagonist bicuculline, and 10 micrograms of the GABAB agonist baclofen or the GABAB antagonist phaclofen were made into conscious animals. Jugular venous blood was collected at 10-min intervals for 3 h, the injection of GABA drug or vehicle was given and samples collected for a further 3 h. The plasma samples were assayed for LH. On completion of the experiments the brains were sectioned to locate the sites of injection. Thirty-four ewes were used, of which 30 had correct guide tube placement. In OVX sheep, both muscimol and bicuculline injection caused suppression of plasma LH concentrations, with a cessation of pulsatile release in many instances. Injection with baclofen or phaclofen into these sheep had no effect on LH secretion. When OVX sheep were treated with 0.5-cm Silastic implants (sc) of E for at least 7 days, there was a variable response to muscimol and bicuculline injection, depending on the degree of suppression of LH secretion by E. When E had little effect on plasma LH levels, muscimol and bicuculline both suppressed LH secretion in a similar manner to that observed in OVX ewes. When E treatment fully suppressed plasma LH levels, muscimol and bicuculline both stimulated LH secretion. This stimulation often took the form of a sustained, nonpulsatile secretion of LH. Baclofen injection into OVX ewes treated with E increased mean plasma LH levels through an increase in pulse amplitude, although there was only an effect on the amplitude of the first pulse after injection. Conversely, phaclofen injection in OVX ewes treated with E resulted in a reduction in LH pulse amplitude. These results provide evidence for a seasonal shift in the regulation of GnRH secretion by GABAergic neurons in the MPOA of the ewe. In contrast to the breeding season, where there was no effect of GABAB ligands, these appear to function in non-breeding season, this may be part of the mechanism for the seasonal shift in the negative-feedback effect of E on LH secretion.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243319 TI - The murine Niemann-Pick type C lesion affects testosterone production. AB - We have determined the effects of the Niemann-Pick type C (NPC) lesion, which impairs transport of cholesterol from lysosomes, on the androgenic status of male NPC mice. The mice have low serum testosterone levels resulting from decreased testosterone secretion. Testosterone secretion is reduced in NPC mouse testes incubated with 8-bromo-cAMP, 20 alpha-hydroxycholesterol, and pregnenolone compared to testosterone release by normal mouse testes under identical conditions. Ultrastructural examination of testes revealed a paucity of lipid droplets, extensive accumulation of inclusion bodies, and distorted endoplasmic reticulum in Leydig cells of adult NPC mice. The hypoandrogenemia caused systemic deficiencies in NPC mice. Seminal vesicles, a testosterone-responsive tissue, were underdeveloped in NPC male mice. The testosterone-responsive kidney beta glucuronidase activity was also underexpressed. Seminal vesicle mass and beta glucuronidase activity were increased by testosterone treatment of NPC mice. Many hepatic proteins, identified by microsequencing, were also deficient in NPC male mice. Levels of alpha 2-mu-globulin, glutathione S-transferase-pi, carbonic anhydrase-III, and selenium-binding protein increased in normal male mice during puberty, but did not increase in the NPC male mice. Based on the increases in protein expression during puberty, differential expression in males and females, and the reported involvement of androgens in regulating expression of some of these proteins, deficient expression of most of these proteins in male NPC mice appears to result from low testosterone levels. We conclude that a defect in testicular testosterone production in NPC male mice causes a pleiotropic deficiency in androgen-sensitive expression of proteins in various organs. PMID- 8243320 TI - Estrogen does not increase bone formation in growing rats. AB - The purpose of these experiments was to test the hypothesis that estrogen has an anabolic effect on cancellous bone formation in long bones of growing rats. In the present studies, estrogen decreased measurements related to cancellous bone formation, including osteoblast number, fluorochrome-labeled bone perimeter, and incorporation of [3H]proline into osteoid. Steady state mRNA levels for bone matrix proteins and mineral apposition rate were unchanged or reduced. The [3H]thymidine labeling index for osteoblasts was very low and was reduced by estrogen treatment. Finally, estrogen decreased the rate of removal of tetracycline previously incorporated into cancellous bone as well as the growth dependent decrease in cancellous bone area, indicating that hormone treatment reduced net bone resorption. In summary, no evidence was secured to support the hypothesis that estrogen has an anabolic action on bone formation; the results obtained in these studies consistently demonstrated a pronounced inhibitory action of the hormone on bone turnover. PMID- 8243321 TI - Selective administration of nicotine into catecholaminergic regions of rat brainstem stimulates adrenocorticotropin secretion. AB - Nicotine (Nic) is a potent stimulus for ACTH secretion, and this response appears to be mediated by central catecholamine secretion. We have previously shown that fourth ventricular administration of Nic rapidly elevated plasma ACTH levels, that a nicotinic cholinergic antagonist, mecamylamine, instilled into the fourth ventricle inhibited the ACTH response to iv Nic, and that Nic stimulated norepinephrine secretion in the hypothalamic paraventricular nucleus. Thus, the present investigations sought to identify Nic-responsive regions in the brainstem that give rise to ascending catecholaminergic afferents resulting in ACTH secretion. Chronic brain and jugular cannulae were implanted, and Nic (50 nl over 30 sec) was infused into the locus coeruleus (LC), nucleus of the solitary tract (NTS -C2 or -A2 regions), C1, or A1 cell regions of freely moving, adult male rats. Injection of Nic (free base, 0.25-10 micrograms) into either the C2 or A2 region of NTS resulted in a dose-dependent increase in plasma ACTH. In contrast, C1 was unresponsive and A1 only showed responses to the highest doses of Nic (5 or 10 micrograms). In LC, Nic in doses of 2.5 micrograms or higher was required to elevate plasma ACTH. This dose is approximately 10-fold greater than that required in NTS-A2. Finally, mecamylamine (0.25 mg/kg body wt, iv), administered 2 min before Nic, abolished the ACTH responses in both C2 and A2 and significantly reduced the 7-min peak ACTH response in LC (P < 0.05). In summary, microinjection of Nic selectively activated the brainstem regions under investigation, with a rank order of sensitivity to Nic that was NTS-A2 > NTS-C2 > LC > A1 > C1 = cerebrospinal fluid. Therefore, systemically administered Nic appears to activate multiple catecholaminergic brainstem regions that are involved in mediating ACTH secretion. PMID- 8243322 TI - Potential role for insulin and cycloheximide in regulating the intrinsic activity of glucose transporters in isolated rat adipocytes. AB - We examined the hypothesis that insulin stimulation of cellular glucose transport may involve a protein synthesis-dependent regulation of glucose transporter (GTer) activity independent of GTer translocation to the cell surface. Rat adipocytes were isolated, incubated with or without 10 micrograms/ml (36 microM) cycloheximide (CHX) for 60 min, and then with or without 7 nM insulin for 30 min. Glucose transport rates were assessed in intact cells, and both glucose transport rates and GTer levels were assessed in subcellular fractions of membrane vesicles before and after reconstitution into artificial liposomes. GTer functional and intrinsic activities were calculated as the ratio between these transport rates and GTer levels in native and reconstituted membranes, respectively. Insulin increased functional activity by 340% in native plasma membrane (PM) vesicles and intrinsic activity by 60% in reconstituted membranes (from 54 +/- 4 to 86 +/- 4 molecules transported per GTer/sec, P < 0.02). CHX preincubation of cells did not interfere with the insulin effect to stimulate glucose transport rate in either intact cells or in native PMs; it did, however, reduce PM GTer levels by 27-30%, but not affecting those in the intracellular pool. However, CHX additively increased the insulin-stimulated intrinsic activity of PM GTers by 67%. Relative reconstitution efficiencies, assessed by immunoblotting both native and reconstituted membranes against specific antibodies, were similar for GLUT 1 and GLUT 4. Although insulin did not alter this efficiency, CHX slightly decreased it for GLUT 4. Our data suggest that insulin stimulation of glucose transport may involve, as part of its mechanism, modulation of the GTer intrinsic activity. We further hypothesize that CHX effects on increasing this activity state of GTer may involve as yet unknown protein synthesis-dependent regulator(s). PMID- 8243323 TI - Generation of 1-37 and 1-38 forms of adrenocorticotropin by mono- and dipeptidyl serine carboxypeptidase activities in bovine pituitary secretory vesicles. AB - ACTH is a 39-amino acid peptide synthesized in the pituitary as part of the precursor molecule, POMC. Analysis of bovine anterior pituitary homogenates and secretory vesicles revealed that in addition to ACTH-(1-39), ACTH-(1-37) and ACTH (1-38) were also present in the lobe, indicating that carboxyl-terminal processing of ACTH-(1-39) occurred in vivo. Mono- and dipe;tidyl carboxypeptidase activities that cleaved ACTH-(1-39) were detected in bovine intermediate and anterior pituitary secretory vesicle membranes and characterized. The dipeptidyl carboxypeptidase activity liberated ACTH-(1-37) and the dipeptide, Glu-Phe, and the monocarboxypeptidase activity generated, to a smaller extent, ACTH-(1-38) and phenylalanine from ACTH-(1-39). Kinetic studies indicated that the formation of ACTH-(1-37) occurred within minutes, whereas the formation of ACTH-(1-38) occurred within hours. Both enzymatic activities had a pH optimum of 5.5 and a Km of 14-18 microM for ACTH-(1-39), and were inhibited by serine and some thiol, but not metallo- or aspartic protease inhibitors. These unique serine carboxypeptidase(s) may function as a converting enzyme in vivo. PMID- 8243324 TI - Localization and quantitation of expression of two glutamate decarboxylase genes in pancreatic beta-cells and other peripheral tissues of mouse and rat. AB - Glutamic acid decarboxylase (GAD) catalyzes synthesis of the inhibitory neurotransmitter gamma-amino butyric acid. Two homologous forms of GAD encoded by separate genes have been cloned from rat brain, with predicted protein sizes of 67 and 65 kilodaltons. GAD is present outside the brain, and pancreatic islet GAD is believed to be a target of autoimmunity in insulin-dependent diabetes mellitus. However, peripheral expression of the two GAD genes is incompletely characterized. We, therefore, investigated GAD expression in peripheral tissues, including pancreas, of mouse and rat. cDNAs encoding GAD 67 and GAD 65 were cloned from mouse brain and shown to be 95% homologous with the rat sequences. RNase protection assay using specific cRNA probes demonstrated expression of both GAD forms in freshly harvested pancreas and testis. Levels of both GAD mRNAs were greater in rat than mouse pancreas. GAD 67 mRNA was more abundant than GAD 65, and both were localized to islet beta-cells by in situ hybridization. In testis, both GAD mRNAs were localized to spermatocytes. Additionally, GAD 67, but not GAD 65, mRNA was detected in mouse and rat spleen and mouse liver. Thus, both GAD genes are expressed in peripheral tissues, with GAD 67 mRNA being more abundant under physiological conditions. The expression of both GAD 67 and GAD 65 genes specifically in islet beta-cells indicates that both GAD forms are candidate autoantigens in rodent models of insulin-dependent diabetes mellitus. PMID- 8243325 TI - Steady state organ distribution and metabolism of thyroxine and 3,5,3' triiodothyronine in intestines, liver, kidneys, blood, and residual carcass of the rat in vivo. AB - We have directly measured the relative distribution and steady state pool sizes of T4 and T3 in blood and extrathyroidal tissues of the whole euthyroid male rat, plus several steady state T3 and T4 metabolism indices in the whole animal, in eight rats infused sc for 7 days with labeled 125I-T3 (T3*) or 125I-T4 (T4*) via implanted minipumps. Liver, kidneys, complete intestine, and residual carcass homogenate extracts and sera were chromatographed on both Sephadex and HPLC. Total tissue T3* and T4* in steady state were assessed and corrected for hormone trapped in residual blood in tissues using radioactive albumin as a vascular marker. Labeled Triac* and/or Tetrac* were prominent among steady state metabolites of T3* or T4* in intestines, feces, and residual carcass, but not in liver or kidneys. Extrathyroidal steady state T3 and T4 pools in residual carcass were 52.8 +/- 5.74 (SD) % of total T3 and 41.2 +/- 4.57% of total T4. The largest extrathyroidal organ pools containing exchangeable T3 and T4 were intestines, with 33.1 +/- 5.46% of total (extrathyroidal) T3 and 18.1 +/- 2.80% of total T4, whereas liver and kidney pools, previously reported as the largest measured, had only 8.0 +/- 1.45% and 2.30 +/- 0.26% of total T3 and 8.72 +/- 1.56% and 1.01 +/- 0.111% of total T4, respectively. Whole blood contained 3.6 +/- 0.385% of total T3 and 31.2 +/- 1.70% of total T4. Also, 21.3 +/- 8.2% of total body T4 was converted to T3 in the whole rat (residual carcass+intestines+liver+kidneys), the production rate of T4 was PR4 = 37.1 +/- 5.28 ng/h.100 g BW and the plasma appearance rate of T3 (PR3min) was 10.6 +/- 2.83 ng/h.100 g BW. T3 production from extrathyroidal T4 was CR3-4 = 6.63 +/- 0.94 ng/h and the minimum T3 secretion rate (SR3min) was 4.02 +/- 0.94 ng/h, each per 100 g BW, indicating that the thyroid secretes more than 38% of whole body T3 in euthyroid steady state. T4 and T3 plasma clearance rates (PCR) were 1.01 +/- 0.29 (T4) and 23.7 +/ 4.36 (T3) ml/h.100 g BW.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243326 TI - Effects of thyroid and glucocorticoid hormones on the level of messenger ribonucleic acid for iodothyronine type I 5'-deiodinase in rat primary hepatocyte cultures grown as spheroids. AB - The activity of type I 5'-deiodinase (5'DI) is known to correlate with thyroid status; it is high in hyperthyroidism and low in hypothyroidism. Recently, it was shown that the increased activity of type I 5'DI in hyperthyroidism is associated with an increase in enzyme contents as well as its mRNA in liver. However, it remains unknown whether thyroid hormone directly regulates the expression of 5'DI mRNA in hepatocytes. In this study, the direct actions of thyroid hormone as well as rT3 and dexamethasone on type I 5'DI mRNA were investigated using primary cultures of rat hepatocytes. Hepatocytes were prepared from euthyroid rats by collagenase perfusion and plated either on collagen-coated dishes for conventional monolayer cultures or on positively charged dishes for spheroid cultures. After hormonal treatments, the levels of mRNAs for type I 5'DI and albumin were determined by Northern blotting. In spheroid cultures, T3 increased type I 5'DI mRNA in a dose- and time-dependent manner, whereas the albumin mRNA level was not altered. A lesser effect was observed in hepatocytes cultured as monolayers. The T3-induced increase in 5'DI mRNA was not inhibited by pretreatment with cycloheximide, indicating that the effect of thyroid hormone on 5'DI mRNA is direct, not requiring de novo protein synthesis. rT3 did not affect the levels in type I 5'DI mRNA increased by T3. On the other hand, dexamethasone alone increased 5'DI mRNA and, when added together with T3, had a synergistic effect. In contrast to T3, dexamethasone increased albumin mRNA. Dexamethasone induced increases in mRNAs for 5'DI and albumin were inhibited by pretreatment of cycloheximide. The present study indicated that T3 increases 5'DI mRNA through a direct action on its gene, whereas the effect of dexamethasone requires de novo synthesis of a protein factor(s). PMID- 8243327 TI - Does alpha-melanocyte-stimulating hormone from the pars intermedia regulate suckling-induced prolactin release? Supportive evidence from morphological and functional studies. AB - Recent evidence suggests that substances derived from the hypophyseal intermediate lobe (IL) play a crucial role in the regulation of suckling-induced PRL secretion. The purpose of the present study was to explore this possibility further by determining whether the suckling stimulus acutely increases the secretory activity of the IL and whether alpha MSH, a major secretory product of the IL, plays a specific role in suckling-induced PRL release. Light microscopic morphometric analysis of serial pituitary sections obtained from lactating rats revealed that as little as 1 min of suckling caused a significant increase in the proportion of the IL that was in secretory configuration (11.8 +/- 0.7% vs. 6.7 +/- 0.5%; 1-min suckled vs. nonsuckled control; mean +/- SE). Moreover, the fraction of the IL in secretory configuration continued to increase after 5 and 10 min of nursing (to 16.0 +/- 0.8% at 5 min and 18.2 +/- 0.7% at 10 min). In contrast, serum PRL was not significantly elevated above the control level after 1 min of suckling (18.1 +/- 13.5 vs. 9.9 +/- 6.5 ng/ml, 1-min suckled vs. control). In fact, a significant rise in PRL levels (to 314.4 +/- 19.4 ng/ml) could be detected only after 10 min of nursing. Thus, secretion by the IL in response to suckling preceded the release of adenohypophyseal PRL, suggesting that a secretory product(s) from the pars intermedia is involved in the modulation of nursing-induced PRL release. Having established a sequential temporal relationship between these two phenomena, we next investigated whether alpha MSH was the IL factor involved in the regulation of suckling-induced PRL secretion. To this end, lactating rats were injected either with antiserum to alpha MSH or preimmune serum and then allowed to nurse their pups. Serial blood samples were taken from the mothers 15, 30, 60, and 90 min after the litters were returned, and serum PRL was measured by RIA. We found that the suckling-induced rise in serum PRL was severely attenuated in animals that received anti-alpha MSH serum. This suppression was most evident at 15 min (70.1 +/- 13.4 vs. 323.5 +/- 127.0 ng/ml, antibody treated vs. preimmune serum control) and persisted throughout the entire 90-min test period. When taken together, our results suggest that suckling-induced PRL secretion is mediated at least in part by alpha MSH released from the hypophyseal IL. PMID- 8243328 TI - In situ ligand binding of recombinant human [125I] activin-A and recombinant human [125I]inhibin-A to the adult rat ovary. AB - Inhibin and activin are hormones produced by ovarian follicles. Specific ovarian cells that bind iodinated recombinant human (rh)-activin-A and rh-inhibin-A were identified by in situ ligand binding. Iodinated rh-molecules were incubated with tissue sections from ovaries, uterus, and oviduct, collected on the mornings of metestrus, diestrus, proestrus, and estrus and the evening of proestrus. Additionally, inhibin/activin subunit mRNA and follistatin mRNA accumulation was examined by in situ hybridization of radiolabeled antisense riboprobes. The cellular site of activin/inhibin binding could thus be colocalized to the cellular site of ligand mRNA and binding protein mRNA production. The association of both [125I]rh-activin-A and [125I] rh-inhibin-A with specific cell types varied across the rat estrous cycle. Nonspecific binding was evaluated by competition with a 1000-fold excess of homologous ligand, and low affinity association of the heterologous ligand was evaluated by competition with a 1000 fold excess of heterologous ligand. [125I]rh-activin-A binding was more widespread than was [125I]rh-inhibin-A binding under our experimental conditions. [125I]rh-Activin-A bound to the granulosa cells of all stages of follicles: unrecruited, growing, and Graafian follicles. Thecal cell binding was found in developing follicles (350-500 microns). The granulosa cells of stimulated follicles (evening of proesterus) bound less [125I]rh-activin-A than those of unstimulated follicles. [125I]rh-Activin-A binding was also associated with antral fluid of follicles in each size class. Although early atretic follicles retained some [125I]rh-activin-A binding, late atretic follicles did not bind [125I]rh-activin-A. Corpus luteum present on metestrus and diestrus bound [125I]rh-activin-A; however, corpus lutea present on proestrus and estrus bound little or no [125I]rh-activin-A. [125I]rh-Activin-A-binding sites were also present in the uterus and oviduct in a cycle-dependent manner. The highest levels of binding were found in the muscle wall of the uterus and the epithelial lining of the thick-walled portion of the uterus on metestrus and diestrus. In addition, [125I]rh-activin-A bound to the cumulus-oocyte complex present in the oviduct on metestrus, but did not bind to the oocytes present in developing follicles. Binding of [125I]rh-inhibin-A was restricted to the antral granulosa cells of 450 to 500-microns follicles. No other ovarian, uterine, or oviduct cells bound [125I]rh-inhibin-A. [125I]rh-Activin-A and [125I]rh-inhibin-A ligand binding was associated primarily with follicles coexpressing inhibin/activin subunit and follistatin mRNA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243329 TI - Identification of a receptor for human mullerian inhibiting substance. AB - Mullerian inhibiting substance (MIS), a Sertoli cell-derived glycoprotein and member of the transforming growth factor-beta supergene family, plays a key down stream role in mammalian sex determination. Identification of a receptor for MIS has now been achieved in a MIS-responsive human vulvar carcinoma cell line, A431, using fluorescein isothiocyanate labeling of recombinant human MIS (FITC-rhMIS) and RRAs with iodinated carboxy-terminal rhMIS. Confocal fluorescence microscopy of A431 cells incubated on ice with 30-nM concentrations of covalently bound FITC rhMIS reveals specific punctate cell surface fluorescent signal. Cytosolic fluorescent signal is seen after incubation at 37 C for 1 h as well as occasional apparent perinuclear accumulation. FITC-rhMIS coincubated with molar excesses of unlabeled rhMIS in A431 cells eliminates cell surface and cytosolic fluorescent uptake. Double label experiments with FITC-rhMIS and tetramethyl rhodamine isothiocyanate epidermal growth factor establish separate binding of each ligand, displaceable, respectively, by large molar excesses of unlabeled rhMIS or epidermal growth factor. RRAs reveal a single, high affinity (Kd, 5.8 nM), saturable, low abundance binding species for carboxy-terminal rhMIS. Solubilized supernatants of A431 whole cells cross-linked with 125I-carboxy-terminal rhMIS identify a band with a mol wt of 88,000 on electrophoresis and autoradiography. This identification of a MIS receptor in A431 cells now permits the design of affinity purification protocols using rhMIS, followed by direct protein microsequencing. PMID- 8243330 TI - Novel self-regulation of human chorionic gonadotropin biosynthesis in term pregnancy human placenta. AB - Term pregnancy human placenta contains hCG/LH receptor mRNA transcripts and immunoreactive receptor protein. Both the receptor transcripts and receptor proteins are present only in trophoblasts. These findings led us to investigate whether hCG can regulate its own synthesis in term pregnancy human placenta. Treatment of placental tissue in static cultures or in a dynamic superfusion system with increasing concentrations of highly purified hCG provoked a biphasic effect on the steady state hCG subunit mRNA levels. Although low concentrations of hCG (< 200 mIU/ml) were not effective, moderate concentrations (200-1000 mIU/ml) increased, and high concentrations (> or = 5000 mIU/ml) either had no effect or actually decreased mRNA levels relative to the control values. This response was specific, because none of the hCG concentrations tested had any effect on glyceraldehyde-3-phosphate dehydrogenase or beta-actin mRNA levels. The effects of hCG on steady state hCG subunit mRNA levels were paralleled by corresponding changes in tissue hCG protein levels. Endogenous hCG appears to down-regulate alpha-subunit mRNA levels and hCG secretion. The hCG effect is probably receptor mediated, because a receptor antagonist, deglycosylated hCG, partially antagonized the hCG action. Treatment with exogenous hCG also down regulated its own receptor mRNA and receptor protein levels. hCG regulation of its alpha-subunit and receptor levels involved both transcriptional as well as posttranscriptional mechanisms. In summary, this is the first demonstration of hCG regulating its own synthesis in term pregnancy human placenta. The findings of this study could offer a potential molecular explanation for the profile of hCG levels in normal pregnant women. PMID- 8243331 TI - Serotonin secretion from rat Leydig cells. AB - In rat Leydig cells, serotonin (5HT) binds to 5HT2 receptors and stimulates the secretion of CRF which in turn acts as an inhibitor of gonadotropin-induced cAMP formation and androgen production. In the present study we defined the regulation of 5HT secretion in cultured Leydig cells. Adult Leydig cells secreted considerable quantities of 5HT (100-150 pg/10(6) cells per 10 min). The release of 5HT was acutely stimulated by hCG (ED50, 1.1 pM) with maximal stimulation at 10 pM hCG (160%). Forskolin also increased (+220%) 5HT release from cultures (ED50, 50 nM) while TPA was much less effective (+20%), indicating a major role for cAMP in gonadotropin-induced 5HT release. This was confirmed by the finding that 8-Br cAMP (1 mM) was an effective stimulus of 5HT release (+360%). Similar increases of 5HT release by hCG were observed in the absence of extracellular Ca2+. However, ionomycin was a potent stimulus of 5HT release, indicating that elevation of cytoplasmic [Ca2+] could also induce amine secretion. The 5HT content of Leydig cells ranged from 300 to 350 pg/10(6) cells, and decreased during stimulation of 5HT release. Also, immunohistochemical studies revealed specific staining of 5 HT in interstitial cells of the adult rat testis. These studies demonstrated that rat Leydig cells contain and secrete 5HT, and that 5HT release is stimulated by gonadotropin acting primarily through a cAMP-mediated mechanism. PMID- 8243332 TI - Evidence for a novel adenylyl cyclase in human epididymal sperm. AB - Since cAMP is considered to play a major role in the acquisition of maturation and fertilizing capacity of mammalian sperm, we investigated the expression of cAMP-synthesizing adenylyl cyclase (AC) in sperm retrieved directly from the human epididymis. Particulate fractions were prepared from purified epididymal sperm samples and AC was monitored by the direct conversion of ATP into cAMP. We report that in great contrast to human ejaculated sperm and other mammalian sperm cells, the human epididymal sperm do not express a Mn(2+)-sensitive AC. However, a functional AC was readily detectable in these sperm cells in the presence of saturating concentrations of Ca2+ (50mM) and bicarbonate (HCO3-, 50mM), a combination that causes maximal activation in human ejaculated sperm. Using these conditions, human epididymal sperm AC showed similar capacity to generate cAMP compared to human ejaculated sperm AC. When assays were performed in the presence of Mg2+ and a saturating concentration of GMP-P(NH)P (50 microM), the hydrolysis resistant GTP analog, and forskolin (100 microM), no activity was detected indicating that the epididymal sperm AC differs from that in somatic cells. These data demonstrate that human epididymal sperm contain an AC that is unique and different from the enzyme system described in somatic cells and other mammalian sperm cells, including human ejaculated sperm. PMID- 8243333 TI - Cytokine-induced C-type natriuretic peptide (CNP) secretion from vascular endothelial cells--evidence for CNP as a novel autocrine/paracrine regulator from endothelial cells. AB - We previously demonstrated that C-type natriuretic peptide (CNP), originally isolated from the porcine brain, is produced by endothelial cells and proposed that CNP can exert local control over vascular tone and growth as a local regulator from endothelial cells. Since cytokines play pivotal roles in the control of vascular tone and structure, we have examined effects of various cytokines on CNP secretion from endothelial cells using the specific radioimmunoassay for CNP. While interleukin (IL)-2 had no significant effect on CNP secretion, IL-1 alpha, IL-1 beta and tumor necrosis factor (TNF)-alpha stimulated CNP secretion in a time- and dose-dependent manner. Among them, TNF alpha, one of the key mediators for inflammation and vascular remodeling, induced more than two orders of magnitude increase in CNP secretion. In addition, lipopolysaccharide (LPS) potently stimulated CNP secretion. These results indicate that IL-1, TNF-alpha and LPS, the endotoxin itself, can regulate local vascular tone and growth through the activation of CNP secretion from endothelial cells. Therefore, CNP could be of clinical relevance as an autocrine/paracrine regulator from endothelial cells for systemic and local cytokine-associated disorders, such as endotoxin shock and atherosclerosis. PMID- 8243334 TI - Autocrine induction of c-fos expression in GT1 neuronal cells by gonadotropin releasing hormone. AB - Activation of GnRH receptors in GnRH neuronal (GT1-7) cells causes marked and transient increases in c-fos expression, with peak response at 30 min. GnRH and des-Gly10-[D-Ala6]GnRH N-ethylamide induced concentration-dependent c-fos responses with EC50s of approximately 0.1 and approximately 1 nM, respectively. GnRH action was mimicked by phorbol 12-myristate-13-acetate (PMA), but stimulation of Ca2+ entry by K(+)-induced depolarization and Bay K 8644 was much less effective. In protein kinase C-depleted cells, the c-fos response to GnRH was reduced to that elicited by increased Ca2+ entry, and the effect of PMA was abolished. Thus, GnRH-induced c-fos expression in GT1 cells appears to be mediated predominantly by protein kinase C, and to a lesser extent by Ca2+. These findings demonstrate that c-fos expression can be induced in a peptidergic neuron by activation of receptors for its neurosecretory product. It is possible that the expression of c-fos in GnRH hypothalamic neurons during the proestrous surge of gonadotropins could likewise be stimulated by a positive feedback action of GnRH on its neuronal receptors. PMID- 8243335 TI - Expression of the type II activin receptor gene in the human placenta. AB - Activin has been suggested to be an autocrine/paracrine regulator in the human placenta. In the present study, we examined the expression of the gene encoding activin type II receptor (ActRII) in this tissue. Using primers corresponding to the published sequence of human ActRII, a 456bp fragment was obtained from cDNAs prepared from the placenta, as well as the ovary and brain, by polymerase chain reaction (PCR). Southern blot hybridization of the PCR products and DNA cloning and sequencing confirmed that the product is the authentic ActRII. Trophoblast cells prepared from both first trimester and term placentae expressed the ActRII gene. When trophoblast cells from term placenta were separated into syncytiotrophoblast- and cytotrophoblast-enriched fractions and incubated for 1-6 days, ActRII gene expression was observed in both cell preparations, with the syncytiotrophoblast-enriched fraction having higher levels of expression at days 1, 3, and 4. These results provide the first direct evidence that the activin type II receptor mRNA is present in human trophoblast cells and strengthen the hypothesis that activin is an autocrine/paracrine regulator of placental function. To our knowledge, this is also the first report that the ActRII gene is expressed in the human brain and ovary. PMID- 8243336 TI - Multiple copies of the bone-specific osteocalcin gene in mouse and rat. AB - The osteocalcin (OC) gene was initially described as a single copy gene encoding the bone specific vitamin K dependent and vitamin D regulated protein. We report here the presence of multiple copies of the gene in mouse and rat. Southern blot analysis and restriction mapping of genomic DNA from several strains of mice indicated the presence of at least three copies of the OC coding sequence within a 19 kb fragment. Two closely linked OC genes contain the proximal promoter region with intact coding sequences. The third potential OC gene includes a 3.5 kb insert between an OC promoter-like region and a coding region that has several amino acid substitutions distributed among functional domains when compared with the normal gene. The 940 nucleotides upstream of the modified coding region lack the well defined 5' regulatory elements that support basal and hormone-responsive transcriptional control. In rats either one or more OC genes were observed in different strains or in Sprague Dawley rats obtained from different suppliers. PMID- 8243337 TI - Induction of Fos-like immunoreactivity in the rat brain following Pasteurella multocida endotoxin administration. AB - The functional neuroanatomy of the immune system link to the CNS was investigated by assessing neuronal activity with Fos immunohistochemistry following systemic lipopolysaccharide (LPS) administration. Two hours after LPS robust Fos-like immunoreactivity (Fos-IR) was observed in several nuclear groups in the brain including the paraventricular and supraoptic nuclei of the hypothalamus, central nucleus of the amygdala, and nucleus of the solitary tract. A similar but diminished pattern of Fos-IR was present at 6 hours and was absent 24 hours after LPS administration. Investigation of the functional neuroanatomy of the acute phase reaction could prove to be critical in enhancing the ability of individuals to combat insults such as tissue damage and inflammation. The central nervous system (CNS), particularly the hypothalamus, is intimately involved in the coordination of the various aspects of the acute phase reaction (reviewed in 1). Understanding the functional neuroanatomy by which the brain responds to immune system challenges would greatly augment the ability to control the deleterious and enhance the beneficial aspects of the acute phase reaction. In this study we have used lipopolysaccharide (LPS or endotoxin) administration as an experimental model to study immune system activation. LPS is a complex glycolipid and a component of the outer membrane of most Gram-negative bacteria (2). Administration of LPS has been demonstrated to induce the secretion of several proteins including interleukin-1 (IL-1), tumor necrosis factor (TNF), and interleukin-6 (IL-6; reviewed in 3). Further, it has been hypothesized that LPS induction of IL-1 and TNF is the key event in the pathogenesis of Gram-negative bacterial septic shock syndrome (2). Many recent studies have utilized immunohistochemistry for Fos, the product of the immediate early gene c-fos, as a marker of neuronal activation. Fos is a nuclear-binding protein that is expressed at increased levels in activated neurons (4). Although the exact function of Fos in the CNS is still unknown, it is thought that Fos is transcribed after cellular stimulation as a means to convert a stimulus into long-term genetic action (for reviews see 5,6). This study investigated the activation of the CNS by peripherally administered LPS isolated from the bacterium Pasteurella multocida. As a marker of neuronal activation, immunohistochemistry for the Fos protein was performed and image analysis was utilized to quantify the Fos induction in the CNS. PMID- 8243338 TI - Cloning and functional expression of a rat brain corticotropin releasing factor (CRF) receptor. AB - Corticotropin releasing factor (CRF), a key neuroregulator of the hypothalamic pituitary-adrenal cortical axis, also displays a broad range of effects on the endocrine, central nervous and immune systems. Having recently characterized the human pituitary CRF receptor by expression cloning of cDNA from a human Cushing's corticotropic adenoma, we report here the structure of the cDNA for a rat brain CRF receptor (rCRF-R) which was cloned by hybridization from a rat brain cDNA library. The sequence of the rCRF-R encodes a 415 amino acid protein comprising seven membrane spanning domains. The rCRF-R is 97% identical at the amino acid level to the human pituitary tumor CRF receptor, differing by only 12 amino acids. When expressed in COSM6 cells, the rCRF-R binds CRF with high affinity (Kd = 1.7 (0.8-3.8)nM). The receptor transduces a CRF stimulated accumulation of intracellular cAMP which is inhibited by the CRF antagonist, alpha helCRF(9-41). These results suggest that the brain expresses a CRF receptor similar to that in the pituitary. PMID- 8243339 TI - Nuclear co-localization of prolactin and the prolactin receptor in rat Nb2 node lymphoma cells. AB - Previous studies have indicated that prolactin (PRL) interacts with specific, high affinity, immunoreactive binding sites within isolated rat hepatocyte nuclei. Moreover, endogenous PRL appears to be bound to this site. However, it remained important to demonstrate nuclear PRL receptors and hormonal translocation in an intact cell system. Therefore, we sought nuclear translocation of PRL and its receptor in the nucleus of PRL-dependent Nb2 node lymphoma cells. Utilizing immunofluorescence (IF) microscopy, growth-arrested cells were found to constitutively express the PRL receptor in the nucleus and in the membrane/cytosol compartments. Addition of PRL stimulated rapid hormone internalization followed by translocation to the nucleus within 6-12 hrs. The translocation of PRL was found to be reversible and dependent upon ATP. These results indicate that an early event coupled to the mitogenic action of PRL in Nb2 cells is hormone transport to the nucleus during the G1 and S phases of cell cycle. Once in the nucleus, PRL bound to its receptor may directly influence gene transcription. PMID- 8243340 TI - Resistance to vertical fracture of roots, previously fractured and bonded with glass ionomer cement, composite resin and cyanoacrylate cement. AB - A recently developed glass ionomer bone cement (Ionos) may be suitable for bonding vertically fractured teeth, but the resulting resistance to repeated fracture of the bonded teeth is unknown. The purpose of this study was to assess in vitro the resistance to repeated fracture of roots which were previously fractured and bonded with Ionos cement and other bonding agents. Thirty-six roots were separated at the cervix and grooved on the coronal surface. Each root was mounted in an Instron machine and a vertical force was exerted until fracture occurred. Both the force required to fracture the root (F) and the root surface area (A) were recorded, and the root halves were bonded with one of the following materials: Ionos bone cement without dentin etching, Permabond 910 cyanoacrylate adhesive, and Gluma bonding system. The bonded roots were maintained in a wet sponge. One week later the roots were re-fractured in the Instron machine, and the forces were recorded (RF). Each one of F, RF, A, and the ratios RF/F and RF/A were compared statistically among the three experimental groups. Overall, RF was at the most 20% of F. Statistically significant differences were obtained only in RF, RF/F, and RF/A (p < 0.0003). Roots bonded with Ionos cement demonstrated significantly lower values than roots bonded with Permabond and Gluma, both of which did not differ significantly. It was concluded that roots bonded with Ionos cement were the least resistant to re-fracture, due to a lesser bonding strength of Ionos bone cement as compared with Permabond and Gluma. PMID- 8243341 TI - Investigation of the children referred to a dental hospital with avulsed permanent incisor teeth. AB - The management of children referred to a dental hospital because of avulsed permanent incisor teeth was considered in a retrospective study. The group consisted of 49 children between the ages of 6 and 14 years. Most of the children (60%) had their avulsed teeth stored dry while only 7% had them stored in milk, and 19% in saliva. Following avulsion the majority of the children attended a dental surgeon or general hospital. Twenty-three children contacted health care personnel within 30 minutes but only 13 had their teeth replanted in this time period. Thirty-six children had 46 incisors replanted. No relationship was demonstrated between the place or personnel who replanted the teeth, and a successful outcome. PMID- 8243342 TI - Bonding of enamel-dentin crown fragments after crown fracture. An experimental study using bonding agents. AB - Previous experimental studies into the use of dentin bonding agents for reattachment of enamel-dentin crown fragments have demonstrated fracture strength about 50-60% of that of intact teeth. In the clinic this has been reflected in the frequent need of repeated bonding of the treated incisors due to refracture. Recently newer bonding agents have been developed which can bond equally well with enamel and dentin. Employing the same experimental model, these bonding agents (All-Bond 2, Scotchbond MP) were used to reattach crown fragments to the remaining portion of sheep incisors. Loading of teeth bonded with these agents in an Instron testing machine at a speed of 1 mm/min demonstrated similar fracture strengths as those previously achieved with Gluma, Scotchbond 2 or Tenure. A second study was carried out whereby fragments were bonded with Scotchbond MP and loaded at various speeds. The results demonstrated that the fracture strength decreased exponentially with increased loading speed. By extrapolation, it was concluded that the strength was nil at a loading speed of approximately 2 m/min. It was concluded that the weak link in the bond between tooth fragment and remaining tooth structure is the bonding resin. Theoretically, a resin which is slightly elastic might act as a shock absorber to withstand functional stress. However, studies carried out with resins of varying moduli of elasticity gave disappointing results probably due to their low flexural strength.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243343 TI - Comparison of the guarding capacities of mouth protectors. AB - The guarding capacity of seven mouth protectors (Erkoflex 3.0 and 4.0, Erkoloc 3.0, 4.5 and 5.5, TranSheet/Perform and TranSheet/LiteLine) was tested mechanically on 20 plaster models, two or three tests on each mouth protector. Two tests on a plaster model without the protector served as control. An appliance was constructed to simulate the impact of an ice-hockey puck on the teeth, the plaster model was fixed onto the device and the minimum force needed to break the plaster teeth under the protector was recorded. The average total thickness and that of the soft and hard layers of the labial plates of each mouth protector were recorded along with the average thickness of the cervical and incisal hard and soft layers. The results showed that the best protection was achieved with the TranSheet/LiteLine model, followed by TranSheet/Perform and Erkoloc 5.5 and 4.5 mouthgards. All these had a resilient layer against the teeth, and it was shown in stepwise regression analysis that the only property having a statistically significant effect on the guarding capacity was the thickness of this cervical soft layer. PMID- 8243344 TI - Long-term retention of a paper point in the periapical tissues: a case report. AB - A case is reported of a 15 mm paper point found in the periapical area of a maxillary right central incisor during endodontic surgery for an unresolved periapical radiolucency. The paper point had been present for between five and nine years. Histopathological examination showed the presence of chronically inflamed fibrous connective tissue with a number of foreign body giant cells and focal collections of chronic inflammatory cells. SEM comparison of the specimen with a similarly treated fresh paper point showed no indication of degradation of the paper point by host defense cells. PMID- 8243345 TI - Implants in children: a case report. AB - A post-crowned, root-filled tooth in a 13-year-old child which is retraumatised resulting in palatal fracture of the root often necessitates removal of the root. The future treatment plan involves appropriate space management and restorative treatment. In this instance the root was retained in order to preserve alveolar bone until the patient was old enough for an implant. PMID- 8243346 TI - Spirochaetes in oral infections. AB - Oral spirochaetes, which are small-, medium- or large-sized, include species of the genus Treponema, many of which have not yet been cultured. They are found in root canal infections, pericoronitis, gingivitis and periodontitis, constituting up to 10% of the flora in endodontic abscesses, 30% in acute necrotizing ulcerative gingivitis, and 56% in advanced marginal periodontitis. The strong proteolytic activity of these organisms probably make them causes of infection rather than consequences. Being able to penetrate tissue, they bring their enzymes, metabolic products, and endotoxins, in direct contact with target cells. This may perturb essential functions of host cells and immunoglobulins. Enzyme activities may also help fulfil the complex growth requirements of spirochaetes in vivo. Reaction between infected periodontal tissue and monoclonal antibodies to Treponema pallidum has suggested that uncharacterized pathogen-related oral spirochaetes have surface structures and functions analogue to this well recognized pathogen. This warrants a more intensified search for the role of spirochaetes in oral infections. PMID- 8243347 TI - Antibacterial effects of various endodontic irrigants on selected anaerobic bacteria. AB - This study was undertaken to determine the antibacterial effects of various endodontic irrigants against six selected anaerobic bacteria. The organisms were mixed separately with dilutions of each of the irrigants in tubes containing fluid thioglycollate broth and allowed to remain in contact for specific time periods before being evaluated for growth. Of the six irrigants tested, chlorhexidine appeared to be the most effective antibacterial substance. Hydrogen peroxide, sodium hypochlorite and REDTA were less effective while the calcium hydroxide and saline solutions proved to be totally ineffective. PMID- 8243348 TI - Convulsions induced by methyl beta-carboline-3-carboxylate in mice: effects of preceding saline injections. AB - The convulsant effects of a high (5 mg/kg intraperitoneally, i.p.) dose of benzodiazepine (BZD) receptor ligand methyl beta-carboline-3-carboxylate (beta CCM), whether or not preceded by administration of two lower doses of beta-CCM (0.5 and 1 mg/kg i.p.) or of saline were studied in nine inbred mouse strains. In five of the strains (A/J, BALB/cBy, C3H/HeJ, CBA/H, and DBA/2J), neither saline nor preceding injections of beta-CCM had any effect on subsequent reactivity to the subsequent convulsant dose. In the other 4 strains, such injections induced either tolerance (CPB-K, NZB), or sensitization (C57BL/6J, XLII), whatever the compound subsequently administered (beta-CCM or saline). In these strains, the data rule out any tolerance or sensitization effect due to beta-CCM, but suggest that such effects could be due to injection itself. PMID- 8243349 TI - Some aspects of prognosis in the epilepsies: a review. AB - The traditional view that epilepsy is usually a chronic condition in which the prognosis is consistently poor has been challenged in the last 2 decades. Evidence from population-based studies and from intervention studies in newly diagnosed patients has produced a wealth of information of a much better prognosis. It is now generally accepted that as many as 70-80% of people developing seizures for the first time will eventually achieve terminal remission, whereas the remaining 20-30% will continue to have recurrent seizures despite all treatment. Despite the high recurrence rate after a first epileptic seizure, remission usually occurs early and for most persons, epilepsy is a short lived condition. The exact role of antiepileptic drugs (AEDs) in this good outcome, however, remains open to debate, because the natural history of the untreated condition is largely unknown. In this article, factors that may influence the prognosis of the epilepsies, including the problems of diagnosis, are reviewed. Special emphasis is given to the issue of spontaneous remission and the question of prognosis of different epileptic syndromes. PMID- 8243350 TI - Knowledge, attitude, and practice toward epilepsy among rural Tanzanian residents. AB - Before a health education program can be established, one must first know what the target population believes and does with respect to the disease in question. Therefore, we performed a study among Tanzanian rural inhabitants to identify their knowledge, attitude, and practice (KAP) toward epilepsy: 3,256 heads of households (mean age 40.2 years, range 15-90 years; M/F ratio 1:1) were interviewed. Of the respondents, 32.9% said they had never seen a seizure; 67.7% said they did not know the cause of epilepsy; 33.3% mentioned various causes including heredity, witchcraft, infection of the spinal cord, hernia; 40.6% believed epilepsy was infectious through physical contact, flatus, breath, excretions, sharing food; 36.8% believed epilepsy could not be cured and 17.1% believed it could not even be controlled; 45.3% believed epilepsy could be treated by traditional healers, and only 50.8% believed hospital drugs were of any use; and 62.7% of the respondents would not allow an epileptic child to go to school for various reasons, including mental subnormality (54.0%), fear of the child falling while alone (65.9%), and fear that the epileptic child would infect other children (11.2%). Concerning what is to be done when a seizure occurs, 33.5% of the respondents would keep away and not touch the person; 16.5% would take some potentially harmful measure such as forcing a mouth gag or forcing a drink such as water (1 even mentioned urine); 5.2% would take unnecessary measures such as rushing the patient to a hospital. Only 35.7% of respondents would perform at least some of the currently recommended first-aid measures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243351 TI - Effect of anticysticercal treatment on the prognosis of epilepsy in neurocysticercosis: a pilot trial. AB - Epilepsy secondary to active or inactive neurocysticercosis (NCC) is a major public health problem in Latin American countries. In an open-label pilot trial, we evaluated and followed (mean = 13 months) 16 patients with epilepsy resulting from active NCC which was treated with anticisticercal (ACC) drugs. These patients were aged 12-68 years with confirmed active NCC and seizures not controlled by adequate antiepileptic drug (AED) therapy. Patients were treated with albendazole or praziquantel (ALB and PZQ, ACC drugs) and AED monotherapy. The number of NCC cysts was markedly reduced by ACC therapy. Thirteen patients remained seizure-free and 2 had only one seizure during follow-up. Our data suggest a positive effect of ACC treatment on the prognosis of epilepsy caused by active NCC, but a prospective, double-blind, controlled study with long-term follow-up must be performed to determine whether ACC therapy improves long-term seizure control. PMID- 8243352 TI - Prevalence of epilepsy in Kelibia, Tunisia. AB - A door-to-door survey was made in Kelibia, Tunisia to determine the prevalence of major neurologic disorders, including epilepsy. The survey was made according to a World Health Organization (WHO) protocol (1981). All individuals responding positively to the screening tool were examined by a neurologic team using well defined diagnostic criteria. One hundred forty-one individuals, alive on prevalence day (July 1, 1985), were identified as having active epilepsy, giving a crude prevalence ratio of 4.04 per 1,000 and an age-adjusted (on WHO population) prevalence ratio of 3.64 per 1,000. Prevalence ratios increase with age (in children and young adults with the highest prevalence ratio at approximately 20 years) and decrease after 40 years. The most frequently identified type was generalized convulsive seizures (93%). The most frequently associated conditions were cerebral palsy and mental retardation. PMID- 8243353 TI - Status epilepticus related to alcohol abuse. AB - We reviewed the case records of 249 adult patients with generalized convulsive status epilepticus (SE) examined in San Francisco General Hospital between 1977 and 1989 and identified 27 patients (10.8%) in whom alcohol abuse was the only identifiable precipitating cause of SE. In 12 patients (44% of the study group), SE was the first presentation of alcohol-related seizures. Seizures with focal features were observed in 11 patients (40.1%), but there was little correlation with localized computed tomography (CT) or EEG abnormalities. SE was controlled with phenytoin (PHT), with or without a benzodiazepine (BZD), in 18 patients (66.7%). Twenty-two patients (81.5%) were discharged with no new neurologic deficits, but time to recovery of baseline mental status was prolonged (> 12 h) in 24 patients. With regard to alcohol abuse history, study patients did not differ from a comparison group with isolated alcohol withdrawal seizures. The results indicate that alcohol abuse is a common cause of SE and that SE may be the first presentation of alcohol-related seizures. Furthermore, the outcome of patients with alcohol-related SE compares favorably with that of patients with SE due to other causes, but recovery of these patients may be complicated by a prolonged postictal state. PMID- 8243354 TI - Chronic intractable epilepsy as the only symptom of primary brain tumor. AB - We identified 39 patients with chronic epilepsy (seizures > or = 2 years) proven to have primary brain tumors. These cases represent approximately 12% of the surgery cases for epilepsy in the same period. Mean age of seizure onset was 13.2 years: mean duration before operation was 10.5 years. Thirty-eight of 39 had normal neurologic examination. Twenty-six tumors were temporal, 7 were frontal, 4 were parietal, and 2 were occipital. Nine of 26 (34.6%) of the temporal group had contralateral interictal EEG spikes. Pathology was 15 ganglioglioma, 13 low-grade astroctoma, 4 oligodendroglioma, 2 low-grade mixed glioma, 1 pleomorphic xanthoastrocytoma, 2 dysembryoplastic neuroepithelial tumor, and 1 ependymoma. Postoperative seizure frequency (minimum follow-up 6 months) ranged from 15 to 16 seizure-free auras only in patients with temporal tumors and total gross tumor removal (mean follow-up 28 months) to 0 of 6 seizure-free in patients with extratemporal tumors who underwent subtotal resection or biopsy. PMID- 8243355 TI - Predictivity of plasma prolactin levels in differentiating epilepsy from pseudoseizures: a prospective study. AB - The predictivity of raised plasma prolactin (PRL) concentrations in differentiating seizure from syncopal attack was prospectively assessed in all patients consecutively admitted to the Clinica Neurologica of Brescia, Italy in a 12-month period who fulfilled the criteria for either a seizure or syncopal attack. Postictal plasma prolactin concentration (P1) was assessed as soon as possible after the event. Three further assessments were performed: P2 was sampled 1 h after P1, P3, and P4 were sampled in the morning for the next 2 days. Patients who had had a seizure showed significantly increased P1 concentrations, when P1 was sampled within 60 min of the attack. In seizure patients assessed > 1 h after the event, P1 was not significantly different from either P2, P3, or P4. In patients who had had a syncopal attack, PRL concentration never increased. In patients assessed < or = 60 min after the seizure, cutoff criterion of P1 exceeding by +3 SD the mean calculated on P2, P3, and P4 yielded a positive predictive value of 89% and a negative predictive value of 61%. These findings confirm that plasma prolactin concentration is highly predictive of true epilepsy but barely predictive of pseudoseizures. PMID- 8243356 TI - Psychogenic seizures in old age: a case report. AB - Psychogenic seizures are unusual after age 60 years. A 73-year-old woman had onset of psychogenic seizures at age 69 years. Five to six attacks occurred each month, usually at night, characterized by an initial subjective sensation and headache followed by generalized stiffening and shaking. Continuous EEG-closed circuit television (EEG-CCTV) monitoring clearly showed these episodes to be nonepileptic. Discontinuation of antiepileptic drugs (AEDs) did not increase the frequency or severity of attacks. Epileptiform discharges were not recorded during the awake, drowsy, or sleeping states. Psychiatric evaluation identified significant turmoil in the patient's life and a history of childhood sexual and physical abuse. Psychogenic seizures may begin in old age and should be considered in the differential diagnosis of intractable seizures in the elderly. Predominantly nocturnal occurrence should not exclude the diagnosis. PMID- 8243357 TI - Effect of chronic electrical stimulation of the centromedian thalamic nuclei on various intractable seizure patterns: I. Clinical seizures and paroxysmal EEG activity. AB - Twenty-three patients with various intractable seizure patterns were divided into four groups based on their most frequent seizure type and their clinical and EEG response to chronic electrical stimulation of the centromedian thalamic nuclei (ESCM): group A, generalized tonic-clonic (GTC, n = 9); group B, partial motor (Rasmussen type) (n = 3); group C, complex partial seizures (CPS, n = 5); and group D, generalized tonic seizures (Lennox-Gastaut type) (n = 6). CM were radiologically and electrophysiologically localized by means of stereotaxic landmarks and by thalamically induced scalp recruiting-like responses and desynchronization. ESCM consisted of daily 2-h stimulation sessions for 3 months. Each stimulus consisted of a 1-min train of square pulses with a 4-min interstimulus interval, alternating right and left CM. Each pulse was 1.0 ms in duration at 60/s frequency and 8-15 V (400-1,250 microA) amplitude. Voltage (V), current flow (microA) and impedance (k omega) at the electrode tips were kept constant. A significant decrease in the number of seizures per month and paroxysmal EEG waves per 10-s spochs occurred in group A patients between the baseline period (BL) and the ESCM period. These changes persisted for > 3 months after discontinuation of ESCM (poststimulation period, Post). Post was accompanied by a significant decrease in the number of paroxysmal EEG discharges. A substantial decrease in seizures and paroxysmal discharges was also observed in patients of group B. In contrast, patients of groups C and D showed no significant changes from BL to ESCM and Post periods, except for a significant decrease in the number of seizures in group D patients from BL to Post periods. PMID- 8243358 TI - Effect of chronic electrical stimulation of the centromedian thalamic nuclei on various intractable seizure patterns: II. Psychological performance and background EEG activity. AB - We studied the effect of electrical stimulation of centromedian thalamic nuclei (ESCM) on seizure control and paroxysmal EEG activity in 23 patients. We report the effect of chronic ESCM on psychological performance and background EEG activity of patients with various intractable seizure patterns. In each patient, a simple specifically designed neuropsychological scales (one for adults and one for children) was administered at the end of the baseline (BL), ESCM, and poststimulation (Post) periods; and 14 consecutive EEG recordings during these periods were performed to determine the degree of neuropsychological improvement and the temporal course of EEG changes. A significant increase in psychological scores and the number of background EEG waves per 10 s was noted in groups A (generalized tonic-clonic seizures, GTC), C (complex partial seizures, CPS), and D (generalized tonic seizures) and the total group of patients from BL to ESCM and from BL to Post periods. Group B patients showed a substantial increase (partial motor seizures) during the same periods. Improvement on psychological performance correlated better with age and baseline degree of deterioration than with the nature of the particular psychological improvement in any given subtest. The improvement in EEG background rhythm was most noticeable at the end of ESCM and at the beginning of the Post periods. Complete normalization of neuropsychologic scores and EEG rhythms was rare, but improvement was significant for both. Psychological scores increased from BL 14 +/- 2 to ESCM 21 +/- 2 and Post 23 +/- 2 (normal expected 26), and EEG background rhythm increased from BL 42 +/- 2 to ESCM 62 +/- 2 and Post 54 +/- 2 EEG waves/10 s. (normal expected > 80). PMID- 8243359 TI - Prototype antiepileptic drug clinical development plan. AB - Antiepileptic drug (AED) development has been generally difficult owing to many factors: regulatory requirements for demonstration of efficacy and safety, subject availability, traditional trial designs, and physicians' beliefs about epilepsy and its treatment. The U.S. Food and Drug Administration (FDA) regulations require a new drug to be shown safe and effective for its intended use before it can be marketed. The unambiguous proof required is a formidable hurdle for AED development. We report a recent clinical development plan highlighting innovations in clinical trial design that have addressed these requirements, discuss alternative endpoints, and compare the results of various trial designs at various stages of development. This model clinical development plan includes trials relevant to all three clinically relevant contexts in which an AED might be used: as an adjunct to an existing regimen, as a substitution for much of an existing AED regimen, and as monotherapy. PMID- 8243360 TI - Effects of monotherapy and polytherapy on the blink reflex in epileptic patients. AB - We performed the blink reflex (BR) in 20 normal volunteers, 13 epileptic patients receiving antiepileptic drug (AED) monotherapy, and 13 epileptic patients receiving AED polytherapy. Comparison of R1, ipsilateral and contralateral R2 and VIIth nerve latencies in the three groups showed no statistically significant differences R1 and VIIth nerve latencies among the three groups. There were statistically significant differences between the polytherapy group and the monotherapy and control groups in comparisons of ipsilateral and contralateral R2. There were no significant differences between the monotherapy group and the control group for ipsilateral and contralateral R2. We hypothesized that AED polytherapy might interfere with synaptic transmission in the polysynaptic pathway of the blink reflex, prolonging the latency of R2. These results provide further evidence of the pathophysiologic effects associated with polytherapy in epileptic patients. PMID- 8243361 TI - Seizures with clonazepam: discontinuation and suggestions for safe discontinuation rates in children. AB - Clinical characteristics of clonazepam (CZP) discontinuation seizures were clarified from 17 patients who developed seizure exacerbation after rapid discontinuation, including abrupt discontinuation, reduction, or replacement with nitrazepam (NZP). Safe discontinuation rates of CZP were estimated by comparing these patients with 23 patients without CZP discontinuation seizures. The seizures consisted of status epilepticus or frequent attacks of generalized tonic clonic seizures (GTC) or non-GTC, and the first attack of GTC. Incidence did not differ significantly between CZP-effective cases and CZP-ineffective cases at discontinuation. The reasons for abrupt discontinuation included adverse effects, insufficient efficacy, difficulty in taking CZP owing to pneumonia, or carelessness of the doctors or the families. Seizure exacerbation occurred 1-6 days after CZP discontinuation. Most of the problems improved with readministration of the original or smaller dosages of CZP or of NZP in an amount threefold or more greater than the original CZP dose. Safe discontinuation rates were estimated to be < or = 0.04 mg/kg for abrupt discontinuation, < or = 0.04 mg/kg/week for reduction, and replacement with NZP of 2.5 or more times the original amount of CZP. These safe discontinuation rates were prospectively confirmed by 84 cases of CZP discontinuation in 54 other patients, and no seizure exacerbation occurred with these discontinuation rates. PMID- 8243362 TI - Hematologic manifestations of long-term valproate therapy. AB - Sixty patients receiving long-term valproate (VPA) monotherapy were studied for hematologic side effects. All were patients in a long-term care facility and ranged in age from 2 to 29 years (mean 14.6 years). Twenty developed at least one prominent hematologic abnormality. Thrombocytopenia and macrocytosis were the most common findings. In patients with macrocytosis, platelet counts were inversely related to VPA levels. Serum B12 levels were increased in 51 of the patients. In 12 patients with macrocytosis who were extensively studied, no etiology for the increased MCV could be identified. An increased number of Pelger Huet-like cells was noted in these 12 patients. None of the patients demonstrated hepatic dysfunction. Hematologic toxicity was never severe enough to discontinue therapy and always responded to small decrements in VPA therapy. VPA was discontinued in only 1 patient, owing to poor seizure control. PMID- 8243363 TI - Reduced bioavailability of moisture-exposed carbamazepine resulting in status epilepticus. AB - A 28-year-old man with seizures well controlled with carbamazepine (CBZ) and valproate (VPA) developed generalized convulsive status epilepticus three days after his CBZ tablets became wet during a rainstorm while he was camping out. He continued the CBZ but avoided ingesting obviously disintegrating tablets. The VPA was not exposed to excessive moisture. Previous random CBZ blood levels had been in the range of 9-13 micrograms/ml; at the time of admission to the emergency room the level had decreased to 3.8 micrograms/ml. The VPA level did not change significantly from baseline. The moisture-exposed CBZ tablets were of expected weight but were swollen and enlarged. Analysis of these tablets by USP paddle method 2 demonstrated a mean dissolution of 16% at 60 min as compared with > 80% for fresh CBZ tablets. After final dissolution of the moisture-exposed CBZ tablets, normal quantities of active drug were noted. We propose that poor dissolution of moisture-exposed CBZ tablets results in reduced bioavailability. We urge caution in the packaging and storage of CBZ to avoid exposure to moisture. PMID- 8243364 TI - Effects of carbamazepine on auditory brainstem response, middle-latency response, and slow cortical potential in epileptic patients. AB - Auditory brainstem responses, middle-latency responses, and slow cortical potentials (ABRs, MLRs, SCPs) were recorded in 21 epileptic patients before and during treatment with carbamazepine (CBZ). The peak-latencies, interpeak intervals, and amplitudes were estimated and evaluated statistically. CBZ monotherapy resulted in prolongation of peak latencies of ABR waves I, III, and V as well as of interpeak intervals I-III and I-V. A significant increase in the peak-latencies of MLR components Na, Pa, and Nb and of interpeak intervals V-Pa and Na-Nb was also observed along with the systematic NaPa amplitude reduction. CBZ also prolonged the peak-latencies of SCP components P1 and N1. Based on the obtained results, we suggest that CBZ exerts suppressive influences both on modally specific (lemniscal) and modally nonspecific (extralemniscal) auditory structures. PMID- 8243365 TI - Carbamazepine effects on preoptic GABA release and pituitary luteinizing hormone secretion in rats. AB - In vivo effects of carbamazepine (CBZ) on the neuroendocrine preopticopituitary feedback system were studied by local application of CBZ through a push-pull cannula into the preoptic area and measurement of local effects on gamma aminobutyric acid (GABA) and distant effects on a subsequent biologic response: luteinizing hormone (LH). Perfusion with 8 and 12 micrograms CBZ/ml cerebrospinal fluid caused a reduction in preoptic GABA release and concomitant suppression of plasma LH levels. These results suggest a GABA component to the mechanism(s) of action of CBZ: (a) CBZ reduces extracellular available GABA concentration, and (b) owing to the known inhibitory role of preoptic GABA in pituitary LH secretion, an increase of postsynaptic GABAergic transmission by CBZ itself could be inferred. PMID- 8243366 TI - Phenytoin and/or stiripentol in pregnancy: infant monkey hyperexcitability. AB - A monkey (Macaca fascicularis) model was used to assess infant hyperexcitability after prenatal exposure to phenytoin (PHT, n = 4), stiripentol (STP, n = 5), or PHT + STP (n = 4). Adult female monkeys were equipped with tether systems and stomach catheters so that drug administration could start 1 month before mating and could be continued throughout gestation. During pregnancy, PHT and STP plasma levels were maintained between 4-12 and 4-10 micrograms/ml respectively (for both monotherapy and polytherapy). Infants were separated from mothers at birth and transferred to the University of Washington's (Seattle) Infant Primate Research Laboratory (IPRL) for postnatal care and testing. Data on a hyperexcitability scale were obtained during cognitive testing for visual and cross-modal recognition memory in 13 infant monkeys when they were between 2 weeks and 3 months of age. The data indicated that infants prenatally exposed to PHT, whether alone or in combination with STP, were at increased risk for hyperexcitability (screeching, refusing to attend to stimuli, lack of visual orientation). This was not true of infants prenatally exposed to STP monotherapy (drug group differences, p < 0.05). PMID- 8243367 TI - Comparative study of the anticonvulsant effect of gamma-aminobutyric acid agonists in the feline kindling model of epilepsy. AB - We made a comparative study of the anticonvulsant effect of GABA agonists on feline amygdala or hippocampal kindled seizures. Progabide (PGB) [gamma aminobutyric acid (GABA) receptor agonist 25-100 mg/kg intraperitoneally, i.p.] significantly reduced both the kindled seizure stage and after discharge (AD) duration in a dose-dependent manner. SKF89976A (GABA uptake inhibitor 0.5-2.0 mg/kg i.p.) also significantly reduced the kindled seizure stage. Toxic doses of SKF89976A caused generalized paroxysmal EEG discharges and myoclonus, but AD generation in the kindled focus was suppressed completely. Furthermore, gamma vinyl GABA (GABA catabolic enzyme inhibitor, GVG 50-200 mg/kg i.p.) significantly reduced the seizure stage, while causing prolongation of the AD duration. In contrast, baclofen (selective GABAB receptor agonist, 1 or 5 mg/kg) did not show anticonvulsant effects on any parameters of kindled seizures. Therefore, these GABA agonists, which potentiate the inhibitory function of the GABAA systems, have potent anticonvulsant effects on partial onset and secondarily generalized limbic seizures. PMID- 8243368 TI - Anticonvulsant effects of bretazenil (Ro 16-6028) during ontogenesis. AB - Anticonvulsant action of a new benzodiazepine, bretazenil (Ro 16-6028), was studied in 240 rats in five age groups: age 7, 12, 18, 25 and 90 days. Motor seizures induced by metrazol (pentamethylenetetrazol, PTZ, 100 mg/kg subcutaneously (s.c.) except for 18-day-old rats which received a dose of 90 mg/kg) served as a model. Animals were pretreated with Ro 16-6028 in doses of 0.001-0.1 mg/kg intraperitoneally (i.p.) 10 min before metrazol. Both types of metrazol-induced seizures, minimal (mMS, predominantly clonic with preserved righting ability) and major (MMS, generalized tonic-clonic), were suppressed by Ro 16-6028 in a dose-dependent manner. Major seizures were always more sensitive to Ro 16-6028 than were minimal seizures. The youngest rats exhibited maximal effects of Ro 16-6028 against major seizures. On the other hand, this drug increased the incidence of minimal seizures in 7- and 12-day-old rats, i.e., in age groups in which this type of seizure is rare under control conditions. PMID- 8243369 TI - Effects of a benzodiazepine, bretazenil (Ro 16-6028), on rhythmic metrazol EEG activity: comparison with standard anticonvulsants. AB - A novel anticonvulsant benzodiazepine bretazenil (Ro 16-6028) was studied electrophysiologically in a model of human absence seizures: rhythmic metrazol activity (RMA) in rats. The effects of Ro 16-6028 pretreatment (0.01, 0.05, or 0.1 mg/kg intraperitoneally, i.p.) were compared with those of clonazepam (CZP, 0.02 or 0.1 mg/kg i.p.), valproate (VPA, 200, 300, or 400 mg/kg) and ethosuximide (ESM, 31.25, 62.5, or 125 mg/kg i.p.) in 45 rats with implanted electrocorticographic electrodes. RMA was elicited by an injection of pentylenetetrazol (metrazol, PTZ) in a dose of 40 or 35 mg/kg i.p. The effects of Ro 16-6028 were similar to those of CZP and VPA, i.e., suppression of RMA episodes, an increase in latency and a decrease in number, and total as well as mean duration. On the other hand, ESM differed from these antiepileptic drugs (AEDs) in inability to shorten the duration of RMA episodes. Based on these results, Ro 16-6028 might be predicted to be efficient against human absence seizures. PMID- 8243370 TI - Profile of anticonvulsant activity and minimal toxicity of methyl 4-[(p chlorophenyl)amino]-6-methyl-2-oxo-cyclohex-3-en-1-oate and some prototype antiepileptic drugs in mice and rats. AB - The anticonvulsant and toxic properties of methyl 4-[(p-chlorophenyl)amino]-6 methyl-2-oxocyclohex-3-en-1-oate (ADD 196022), were compared with those of phenytoin (PHT), carbamazepine (CBZ), and valproate (VPA). These compounds were evaluated in mice and rats using well-standardized anticonvulsant testing procedures. Results indicate that ADD 196022 is a very potent anticonvulsant in the maximal electroshock seizure (MES) model. The compound was effective in nontoxic doses after intraperitoneal (i.p.) administration in mice and oral administration in rats. In mice, i.p. administration of ADD 196022 resulted in an ED50 value of 26.2 mg/kg as compared with a value of 6.48 mg/kg for PHT in the same assay. ADD 196022 was more potent that PHT in the oral rat model, having an ED50 value of 5.79 mg/kg as compared to 23.2 mg/kg for PHT. ADD 196022 was ineffective in nontoxic doses against all other seizure models evaluated and thus has a pharmacologic profile similar to that of PHT. PMID- 8243371 TI - Role of the fastigial nucleus in generalized seizures as demonstrated by GABA agonist microinjections. AB - The cerebellum is electrically and metabolically active during seizures. Numerous studies have also shown that cerebellar electrical stimulation and lesions of the cerebellar cortex or nuclei influence seizure threshold, but there are significant contradictions, with different effects observed even in investigations using the same species and similar seizure types and experimental manipulations. Discrete intracerebral microinjection of neuroactive agents has been used to characterize the way in which other brain regions control seizures, but has not been applied to the cerebellar systems. This approach has advantages because effects are restricted to specific receptors and spare passing axons; experimental variables also can be simply specified and reproduced. We used this method to characterize the role of the cerebellar nuclei in seizures and to determine if observed effects could be reproduced with different agents at different doses. Effects of bilateral control microinjections in the fastigial (medial) cerebellar nucleus were compared with different doses of the GABAA agonist piperidine-4-sulfonic acid and the GABAB agonist (-)baclofen (Bf). Soon after injection, the animals were ataxic. After 4 min, seizures were induced by timed continuous intravenous (i.v.) bicuculline (BIC) infusion. Both GABA agonists produced significant reductions in myoclonic, clonic, and tonic seizure thresholds. Injections just dorsal or anterior to this nucleus and bilateral dentate (lateral) nucleus injections had little effect on seizures. These results demonstrate that the cerebellar system does control seizures, but does not provide support for the early concept that cerebellar stimulation and systemic phenytoin block seizures through inhibition of cerebellar nuclei secondary to Purkinje cell activation. PMID- 8243372 TI - Phosphate energy metabolism during domoic acid-induced seizures. AB - The effect of domoic acid-induced seizure activity on energy metabolism and on brain pH in mice was studied by continuous EEG recording and in vivo 31P nuclear magnetic resonance (NMR) spectroscopy. Mice were divided into ventilated (n = 6) and nonventilated (n = 7) groups. Baseline EEG was 0.1-mV amplitude with frequence of > 30-Hz and of 4-5 Hz. After intraperitoneal (i.p.) administration of domoic acid (6 mg/kg), electrographic spikes appeared at increasing frequency, progressing to high-amplitude (0.1-0.8 mV) continuous seizure activity (status epilepticus). In ventilated mice, the [31P]NMR spectra showed that high-energy phosphate levels and tissue pH did not change after domoic acid administration or during the intervals of spiking or status epilepticus. Nonventilated mice showed periods of EEG suppression accompanied by decreases in the levels of high-energy phosphate metabolites and in pH, corresponding to episodic respiratory suppression during the spiking interval. In all animals, status epilepticus was followed by a marked decrease in EEG amplitude that progressed rapidly to isoelectric silence. [31P]NMR spectra obtained after this were indicative of total energy failure and tissue acidosis. In a separate group of ventilated mice (n = 4), domoic acid-induced status epilepticus was accompanied initially by an increase in mean arterial blood pressure (MAP) that slowly returned to baseline level. Isoelectric silence was accompanied by a decrease in MAP to 75 +/- 8 mm Hg. These experiments suggest that domoic acid-induced seizures are not accompanied by an increase in substrate demand that exceeds supply. PMID- 8243373 TI - Innovative designs of controlled clinical trials in epilepsy. AB - Uncontrolled noncomparative clinical observations of investigational antiepileptic drugs (AEDs) often lead to overoptimistic efficacy results and are therefore of very limited value for clinical AED development. The classic add-on trial with placebo as control treatment, in contrast, has provided unequivocal evidence of the efficacy of classic and new AEDs and has also identified less useful AEDs. Drug interactions, carryover effects, difficulty in analyzing individual drug action, and the recognition that monotherapy is by far the more common way of prescribing AEDs have led to the development of classic active control monotherapy trials. A major problem of these trials is a no-difference outcome, which allows no useful interpretation. Recently, two alternative monotherapy designs have been developed to avoid the deadlock of a no-difference outcome. In these designs the active control drug is administered in an attenuated form (low dosage or low concentration) or a placebo control is used when standard treatment is discontinued during presurgical evaluation. Both designs have produced unequivocal evidence of the efficacy of the investigational AED during monotherapy. Ethical concerns are minimized by the introduction of preset escape criteria for patient protection. These designs are valuable new supplements for the clinical development of investigational AEDs for monotherapy in epilepsy. In our opinion, alternative monotherapy designs should be preceded by more than one pivotal add-on, placebo-controlled trial. PMID- 8243374 TI - Felbamate in the treatment of Lennox-Gastaut syndrome: results of a 12-month open label study following a randomized clinical trial. AB - Felbamate (FBM) is a new antiepileptic drug (AED) that has been evaluated in partial seizures and in the Lennox-Gastaut syndrome (LGS). When tested against placebo in an add-on, randomized, double-blind trial in 73 children with LGS, FBM significantly reduced the frequencies of astatic (atonic) seizures and generalized tonic-clonic seizures plus total seizure counts. In addition, FBM treated subjects improved significantly on a parent-rated global evaluation and had fewer injuries. Overall, approximately 50% of subjects experienced a 50% or greater reduction in total seizure frequency and a dose-response relationship was apparent. The improvement that occurred in the double-blind study has been sustained for at least 12 months in subsequent open-label follow-up studies. In the first month of FBM treatment, 62% of the subjects who had previously received placebo had a reduction in total seizure frequency of > 50%. By the 12-month follow-up point, approximately half of the patients had a 50% reduction in total seizure count. Astatic seizures responded even better, with two-thirds of patients having a reduction of > 50% in astatic seizure frequency after 12 months of treatment. Based on adverse experience reports thus far, FBM appears to be well tolerated. FBM is the first drug to be shown effective in the LGS in randomized controlled trials. Although few subjects with LGS became seizure free, the frequency of the most severe seizure types decreased and the patients' global functioning improved. PMID- 8243375 TI - Felbamate in the treatment of refractory partial-onset seizures. AB - Felbamate (FBM) is a new antiepileptic drug (AED) that has been tested in open and controlled studies in patients with therapy-refractory partial-onset seizures. Proof of efficacy is based on results of five controlled studies (three with polytherapy and two with monotherapy). In two of the three polytherapy studies, a classic placebo crossover design was used. The third study used a novel design evaluating the efficacy of FBM in a placebo-controlled parallel design in patients completing an evaluation for epilepsy surgery. The primary efficacy variable in this study was the number of patients who experienced a fourth seizure before the end of the study. Forty-six percent of patients randomized to FBM stopped treatment prematurely because of a fourth seizure compared with 88% randomized to placebo. Two studies investigating the efficacy in monotherapy were performed. Both studies used an identical trial design comparing FBM with a low dosage of valproate (VPA). The efficacy of FBM was found to be superior to the low-dosage VPA for both studies. Open long-term follow-up studies have confirmed the long-term efficacy of FBM for up to 12 months. Overall, FBM was well tolerated in both poly- and monotherapy. PMID- 8243376 TI - Felbamate: successful development of a new compound for the treatment of epilepsy. AB - Felbamate (FBM) is an effective and safe novel antiepileptic drug (AED) for add on treatment in adults with refractory partial seizures as shown in three pivotal controlled trials. In addition, FBM is effective and safe in monotherapy in adults with refractory partial seizures. FBM is also effective and safe as add-on therapy for children and adults with refractory Lennox-Gastaut syndrome. The effective daily dosage is approximately 30-45 mg/kg divided into three or four doses with resulting plasma concentrations of 50-80 mg/L. The safety profile of FBM is limited to mild gastrointestinal complaints, insomnia, and nonspecific CNS symptoms. Six pivotal controlled trials, with both classic and innovative design, showed that FBM is a useful AED. PMID- 8243377 TI - The Lennox-Gastaut syndrome. AB - One of the most challenging areas in nosology is in the field of severe generalized epilepsy of early childhood. This is certainly true in the case of Lennox-Gastaut syndrome (LGS), an age-related epileptogenic encephalopathy which comprises several types of generalized seizures including tonic seizures, atypical absence seizures and frequent status epilepticus. EEG shows generalized slow spike waves, and as the disease progresses, cognitive functions deteriorate. LGS is listed in the 1989 classification of the International League Against Epilepsy alongside epilepsy with myoclonic astatic seizures and West's syndrome. A number of variants or atypical forms have been proposed. As a result, differential diagnosis presents a major challenge and includes specific generalized epilepsies, i.e., metabolic or inflammatory; secondarily generalized epilepsies, i.e., those arising from the frontal lobe; and severe forms of idiopathic generalized epilepsy, i.e., Doose syndrome. Antiepileptic drug (AED) treatment of LGS has been disappointing. Results obtained from anterior callosotomy have been promising, but only a small number of patients have been evaluated. Although the syndrome is rare, the severe nature and intractability of LGS emphasizes the need for the development of specific AEDs which would completely modify the quality of life for these patients. PMID- 8243378 TI - Impact of environment, stress, occupational, and other hazards on sexuality and sexual behavior. AB - Sexual health is important for general as well as reproductive health. The effects of the environment on sexual health are complex, however, because of the psychosomatic nature of human sexuality. The effects of any specific environmental agent on sexual function will therefore be modified or amplified by psychosocial factors, and any assessment of the effects of the agent will need to take those factors into account. As a consequence, we have little direct evidence of the adverse effects of the environment on sexuality. This paper therefore considers a) the aspects of sexuality that may by susceptible to environmental effects and the likely mediating mechanisms. These are considered under three headings: psychophysiological, endocrine, and subjective/interpersonal; b) the types of relevant environmental factors, including toxic pollutants, self administered toxins, diet, situational factors, adversity, stress, and social and cultural factors. As yet research has not controlled adequately for the various confounding factors. It is suggested that a first step is to investigate the effects of work stress (e.g., shift and night work) on sexual relationships, comparing the effects across contrasting cultural settings. Such research would then provide a basis for cross-cultural study of other environmental factors. PMID- 8243379 TI - Effect of radiation on the human reproductive system. AB - Irradiation may have a profound effect on reproductive function. The schedule of the delivered irradiation (total dose, number of fractions, and duration) is an important determinant of the radiobiological effect on the tissues involved and varies among different tissues and organs. Irradiation to the central nervous system may affect the timing of the onset of puberty, result in hyperprolactinemia, or cause gonadotropin deficiency if the hypothalamic pituitary axis is involved in the radiation field. Direct irradiation to the testis will, in lower doses, affect the germinal epithelium: doses of irradiation greater than 0.35 Gy cause aspermia, which may be reversible. The time taken for recovery increases with larger doses; however, with doses in excess of 2 Gy aspermia may be permanent. At higher radiation doses (> 15 Gy), Leydig cell function will also be affected. In addition to radiation dose, the vulnerability of the testis is dependent on the age at irradiation and the pubertal status of the male. In the female, the response of the ovary to the effects of irradiation varies with age as well as dose, and separation of ovarian dysfunction into hormonal and fertility effects is not clearcut. An ovarian dose of 4 Gy may cause a 30% incidence of sterility in young women, but 100% sterility in women over 40 years of age. Pelvic irradiation may also have a profound effect on the uterus, with arrested growth in the prepubertal girl, and failure of uterine expansion during pregnancy with subsequent miscarriages and premature labor. PMID- 8243380 TI - Female reproductive function in areas affected by radiation after the Chernobyl power station accident. AB - This paper reports the results of a comprehensive survey of the effects of the accidental release of radiation caused by the accident at the Chernobyl nuclear power station in April 1986. The accident and the resulting release of radiation and radioactive products into the atmosphere produced the most serious environmental contamination so far recorded. We have concentrated on evaluating the outcomes and health risks to women, their reproductive situation, and consequences for their progeny. We have concentrated on two well-defined areas: the Chechersky district of the Gomel region in Belorussia and the Polessky district of the Kiev region in the Ukraine. A number of investigations were carried out on 688 pregnant women and their babies, and data were obtained from 7000 labor histories of the development of newborns for a period of 8 years (3 years before the accident and 5 years after it). Parameters examined included birth rate, thyroid pathology, extragenital pathology such as anemias, renal disorders, hypertension, and abnormalities in the metabolism of fats, complications of gestation, spontaneous abortions, premature deliveries, perinatal morbidity and mortality, stillbirths and early neonatal mortality, infections and inflammatory diseases, neurological symptoms and hemic disturbances in both mothers and infants, trophic anomalies, and biochemical and structural changes in the placenta. Several exogenous, complicating influences were also considered such as psycho-emotional factors, stress, lifestyle changes, and others caused directly by the hazardous situation and by its consequences such as treatment, removal from affected areas, etc.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243381 TI - Environmental release of chemicals and reproductive ecology. AB - Reproductive ecology is defined as "the study of causes and mechanisms of the effects of environmental risk factors on reproductive health and the methods of their prevention and management." Major areas of concern, within the purview of this paper, relate to adverse pregnancy outcomes, effects on target tissues in the male and the female, and alterations in the control and regulatory mechanisms of reproductive processes. Teratogenic potential of chemicals, released as a result of accidents and catastrophes, is of critical significance. Congenital Minamata disease is due to transplacental fetal toxicity caused by accidental ingestion of methyl mercury. Generalized disorders of ectodermal tissue following prenatal exposure to polychlorinated biphenyls have been reported in Taiwan and Japan. The Bhopal gas disaster, a catastrophic industrial accident, was due to a leak of toxic gas, methyl isocyanate (MIC), in the pesticide manufacturing process. The outcome of pregnancy was studied in female survivors of MIC exposure. The spontaneous abortion rate was nearly four times more common in the affected areas as compared to the control area (24.2% versus 5.6%; p < 0.0001). Furthermore, while stillbirth rate was found to be similar in the affected and control areas, the perinatal and neonatal mortality rates were observed to be higher in the affected area. The rate of congenital malformations in the affected and control areas did not show any significant difference. Chromosomal aberrations and sister chromatid exchange (SCE) frequencies were investigated in human survivors of exposure. The observed SCE frequencies in control and exposed groups indicated that mutagenesis has been induced. Strategies for the management, prediction, and preventability of such disasters are outlined. PMID- 8243382 TI - Impact of the environment on reproduction from conception to parturition. AB - Consideration of environmental influences on human reproduction must include an investigation of the socioeconomic factors that play an important role in embryo fetal development. Nutritional factors are of prime importance, and modern methods of food production and supply have led to new hazards for pregnant women. For example, pregnant women have been advised in a number of European countries not to eat liver due to the very high concentration (frequently exceeding 100,000 IU per 100 g) of vitamin A. The clear demonstration that periconceptional vitamin supplementation can dramatically reduce the recurrence rate of neural tube defects suggests that folic acid deficiency may be common even in otherwise well nourished women. Modern methods of food production manufacture and sale have increased the risks of Salmonella and Listeria infections in pregnancy. The dangers of social drug use, especially alcohol and tobacco, are discussed. The types of adverse effect that can result from chemical exposure during the different stages of pregnancy is reviewed with a discussion of some of the problems of epidemiological studies in attempting to identify toxic chemicals. The extent of the ignorance concerning the reproductive toxicity of industrial and environmental chemicals is emphasized by the fact that data are available on only a small percentage of even the high-production volume (1000 tons/year) chemicals. The evidence of reproductive toxicity from exposure to lead, methyl mercury, polychlorinated biphenyls, polychlorinated dibenzofurans, and hexachlorobenzene are briefly reviewed. Some areas of research needs are discussed with emphasis on nutrition, food production, preparation, and storage and the new problems that may be introduced by genetic manipulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243383 TI - The epidemiology of disasters and adverse reproductive outcomes: lessons learned. AB - A disaster has been defined as a disruption of human ecology that exceeds the capacity of the community to function normally. Little is known about the adverse effects of natural disasters on reproductive outcomes. Important lessons can be derived from several disasters caused by human factors, such as the Minamata Bay disaster. Adverse reproductive outcomes include infertility, early pregnancy loss, stillbirths, congenital malformations, and serious developmental disabilities such as cerebral palsy and mental retardation. Recent disasters like the Chernobyl and Bhopal explosions have provided important lessons on the need for accurate and sound information about the risk of prenatal exposures for adverse reproductive outcomes. To study questions of adverse reproductive outcomes and disasters requires a well-planned approach. It should include early development of surveillance for adverse reproductive outcomes, analytic studies on the risk of disasters from direct and indirect effects, sensitive methods to measure early pregnancy loss, and long-term follow-up programs to assess outcomes such as developmental disabilities. PMID- 8243384 TI - Approaches to evaluating reproductive hazards and risks. AB - Development of approaches to risk assessment for reproductive toxicity has aided in the critical evaluation of the scientific basis for interpretation of data and the description of assumptions underlying the process. In addition, it has helped to standardize, to the extent possible, the use of qualitative and quantitative data in the hazard identification and dose-response processes and to identify research needed to fill gaps in the available database. The standard study protocols for evaluating reproductive and developmental hazards include developmental toxicity studies and both short-term and longer-term reproductive studies. These study protocols have been in use for several decades, but development of risk assessment approaches has prompted the recommendation of additional end point measures to these protocols. These include evaluation of specific neurologic and behavioral function of offspring following prenatal and postnatal exposure, evaluation of sperm production and quality, reproductive organ weights, and more in-depth testicular histopathology in males, as well as measures of age at vaginal opening, vaginal cytology, oocyte toxicity, time to mating, gestation length, and reproductive organ weights in females. Current approaches to risk assessment in reproductive toxicity involve the determination of a no-observed-adverse-effect level (NOAEL) and the application of uncertainty factors (UFs) to account for differences between the experimental animal species and humans, variability in sensitivity within the human population, and other factors as necessary to derive the reference dose (RfD), or a specified RfD for developmental toxicity to account for the short period of exposure required.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243385 TI - Design options and methodological fallacies in the studies of reproductive failures. AB - Reproductive failures are at first sight well suited for epidemiologic research. The time of pregnancy is closely monitored, and failures such as spontaneous abortions and subfecundity are rather frequent. Although epidemiologists' interest in the field has been growing, there is still disappointingly little new information of relevance for prevention. A number of methodologic shortcomings may explain this. A large part of disease classification is not well suited for etiologic research, reduced fertility has diminished the populations at risk, close medical monitoring tends to mask causal links, and many scientific problems related to this area bring limitations to the research field. Still, much more could be learned from a systematic use of epidemiologic knowledge, existing registers, and the joint effort between different research groups. PMID- 8243386 TI - Registries of congenital anomalies: EUROCAT. AB - Congenital anomalies are one of the potential adverse effects of the environment on reproductive health. Registries of congenital anomalies are useful to detect abnormal frequencies, clusters, and trends. Such registries should meet a number of conditions, including an appropriate population denominator, an efficient system for collecting information, standardized diagnostic procedures, postmortem examinations of still-births, and linkage of records. The EUROCAT (European Registration of Congenital Anomalies and Twins) program is a Concerted Action of the Commission of the European Communities initiated in 1979. One of its objectives is the surveillance of congenital anomalies as related to environmental hazards. This surveillance system covers at present 350,000 births per year in 15 countries. A number of problems encountered in the development of EUROCAT and in the course of ongoing activities are reviewed: populations coverage, classification of malformations, coding, definition and coverage of late fetal death, registration of induced abortion, validation of diagnostic information, registration of late diagnosed cases, and maintenance of motivation in data collection. The issue of confidentiality and the need for strict safeguards for the protection of individual privacy are emphasized. PMID- 8243387 TI - Impact of the environment on reproductive health: executive summary. AB - The papers presented at the workshop on the "Impact of the Environment on Reproductive Health" are published in this issue of the EHP Supplements. After the formal presentation of the papers, the authors and scientists met to discuss the important aspects of environmental issues affecting human reproductive health. This Executive Summary was compiled by the organizers and editors of the workshop and the proceedings. PMID- 8243388 TI - A reproductive hazards research agenda for the 1990s. Research Needs Working Group. AB - There is substantial scientific and public concern about the potential effects of occupational and environmental toxicants on reproductive health. These effects include impaired functioning of the reproductive systems of men and women as well as a broad spectrum of developmental problems expressed in offspring. Research on reproduction and development is among the most complex undertakings in biomedical research. This complexity is due in part to the intricate biology of reproduction, the multiple targets involved (male, female, and offspring), the uncertainties in extrapolating from animal models to humans, and the problems involved in accurately characterizing exposures and outcomes in epidemiologic investigations. However, given the relatively brief history of research into toxicant-induced reproductive health effects, we have made enormous strides in our knowledge over the past decade. In particular, recent advances in reproductive biology and biotechnology and in the development of biological markers of exposure, effect, and susceptibility are greatly enhancing our ability to study cause-effect relationships. In this paper, the Research Needs Working Group proposes ways to apply existing knowledge to better protect reproductive health and suggests directions for future research. Fulfilling this challenging agenda will require responsible cooperation by labor, industry, government, individual citizens, and the scientific community. Further research and collaboration are essential to both prevent adverse reproductive and developmental outcomes and to formulate a sound scientific basis for policy making. PMID- 8243389 TI - Occupational and environmental reproductive hazards education and resources for communities of color. AB - Little research has been published on the occupational and environmental hazards affecting people of color. Even less is known about the hazards that affect women of color. Although women of color have always been aggressive participants in the work force, their labor activity has increased dramatically over the last decade. Current job placement patterns suggest that women of color are concentrated in the lowest-paying and most hazardous jobs. In this paper, we specifically focus on occupational and environmental reproductive health concerns. We write with the understanding that reproductive hazards can affect pregnant women, nonpregnant women, and men, as well as the health and development of young children. Emphasis is placed primarily on African American women, because information on Hispanic, Native American, and Asian women is very limited. We discuss the participation of women of color in the labor force, using the U.S. Department of Labor categories. We review specific occupational hazards associated with each category of work and briefly discuss environmental hazards, noting that communities of color are at a disproportionate risk of exposure. Finally, we present the consensus report of the Community Education Working Group from the Woods Hole Conference on Occupational and Environmental Reproductive Hazards. PMID- 8243390 TI - Identifying environmental factors harmful to reproduction. AB - Reproduction is essential for the continuation of the species and for life itself. In biological terms, living and reproducing are essentially one and the same. There is, therefore, no sharp division between identifying factors harmful to reproduction and identifying factors harmful to life or vice versa. Detection of harmful factors requires balanced use of a variety of methodologies from databases on structure-activity relationships through in vitro and in vivo test systems of varying complexity to surveys of wildlife and human populations. Human surveys provide the only assured means of discriminating between real and imagined harmful factors, but they are time consuming and provide information after the harm has been done. Test systems with whole animals provide the best prospects for identifying harmful factors quickly, but currently available methods used for testing agrochemicals and drugs need a thorough overhaul before they can provide a role model. Whether there is a need for new methodology is doubtful. More certain is the need to use existing methodology more wisely. We need a better understanding of the environment--whatever it is--and a more thoughtful approach to investigation of multifactorial situations. PMID- 8243391 TI - Improving education and resources for health care providers. AB - Workers and citizens are turning increasingly to the health care system for information about occupational and environmental reproductive hazards, yet most primary care providers and specialists know little about the effects of occupational/environmental toxicants on the reproductive system or how to evaluate and manage patients at potential risk. Although it is unrealistic to expect all clinicians to become experts in this area, practitioners should know how to take a basic screening history, identify patients at potential risk, and make appropriate referrals. At present, occupational and environmental health issues are not well integrated into health professional education in the United States, and clinical information and referral resources pertaining to reproductive hazards are inadequate. In addressing these problems, the conference "Working Group on Health Provider Education and Resources" made several recommendations that are detailed in this report. Short-term goals include enhancement of existing expertise and resources at a regional level and better integration of information on occupational/environmental reproductive hazards into curricula, meetings, and publications of medical and nursing organizations. Longer term goals include development of a comprehensive, single-access information and referral system for clinicians and integration of occupational and environmental medicine into formal health professional education curricula at all levels. PMID- 8243392 TI - A labor perspective on workplace reproductive hazards: past history, current concerns, and positive directions. AB - The Supreme Court's March 1991 ruling in United Automobile Workers (UAW) versus Johnson Controls barring corporate "fetal protection policies" was a major victory for women's employment rights and has health and safety implications for both sexes. However, 2 years after the Court's decision, the union's work is far from over. The UAW has yet to see what policy Johnson Controls will implement in place of the old one. Formulating solutions to the concerns of workers who are exposed daily to reproductive health hazards on the job will continue to be on labor's agenda. Preventing hazardous exposures is the first priority. This goal would be furthered by setting occupational health and safety standards designed to protect workers' general and reproductive health. Support for the Comprehensive Occupational Safety and Health Reform Act (COSHRA) would also positively affect health and safety in the workplace. Where hazards have not yet been abated, the framework of transfers and income protections for all workers with temporary job restrictions should be examined. The Legal/Labor Working Group convened at the Occupational and Environmental Reproductive Hazards Working Conference authored guidelines for developing a model reproductive hazards policy. These recommendations can serve as a guide for implementation of nondiscriminatory and health-protective policies by employers. PMID- 8243393 TI - Litigating reproductive and developmental health in the aftermath of UAW versus Johnson Controls. AB - In a major decision handed down last term (International Union [UAW] versus Johnson Controls, Inc.), the Supreme Court ruled that employment practices excluding fertile or pregnant women from the workplace because of alleged concerns for fetal health constitute illegal sex discrimination. We analyze the three opinions in the case and explain why the decision was an essential first step to promoting reproductive and developmental health in the workplace. Continued progress toward eliminating or reducing reproductive occupational risks will require comprehensive legal strategies involving private lawsuits, governmental regulation and enforcement actions, and new legislation designed to preserve the existing rights of workers and to obtain new and additional protections. Finally, we caution that, in designing such strategies, it will be important to avoid solutions that either shift responsibility for reproductive health to workers, rather than to employers, or that undermine other important legal rights. PMID- 8243394 TI - Protecting reproductive health and the environment: toxics use reduction. AB - Toxics use reduction is a new chemical hazard management approach that has emerged in several state laws over the past years. While toxics use reduction has been promoted as a means of preventing environmental pollution, little thought has been given to its adoption as a means of managing reproductive hazards. This paper provides illustrations of use reduction approaches to conventionally recognized reproductive and developmental toxicants. These approaches will require the opening of a new dialogue between industrial designers and process managers and those most concerned about reproductive health. Several different strategies are proposed that might be adopted into state programs for promoting reduction in the use of reproductive and developmental toxicants. PMID- 8243395 TI - A critical review of studies of the association between demands for hospital services and air pollution. AB - Studies of the associations between air pollution and hospital admissions and emergency room use are reviewed, including studies of air pollution episodes, time-series analyses, and cross-sectional analyses. These studies encompass a variety of methods of analysis and levels of air quality. Findings from all three types of studies were generally consistent in that almost all of the studies reviewed found statistically significant associations between hospital use and air pollution; this unanimity may have resulted in part from publication bias. These associations were characterized by elasticities of the order of 0.20; i.e., a 100% change in air pollution was associated with a change in hospital use of about 20%, for specific diagnoses. Respiratory diagnoses were emphasized by most studies; cardiac diagnoses were included in five of them. The air pollutants most often associated with changes in hospital use were particulate matter, sulfur oxides, and oxidants. Apart from the major air pollution episodes, there was no obvious link between air pollution level and the significance or magnitudes of the associations. Long-term indicators of hospitalization appeared to also be influenced by medical care supply factors, including the numbers of beds and physicians per capita. These nonpathological causal factors could also have influenced the findings of the time-series studies by introducing extraneous factors in the patterns of admissions. Although consistent associations have been shown between hospital use and air pollution, further research is required to distinguish among potentially responsible pollutants and to deduce specific dose response relationships of general utility. PMID- 8243396 TI - Enhancement of the pneumotoxic effect of cadmium acetate by ionizing radiation in the rat. AB - Experiments involving 120 male Wistar rats were performed to study the effect of treatment with cadmium acetate and external irradiation. A single 0.5 mg/kg body weight dose of cadmium acetate was administered intratracheally. Shortly thereafter, the animals received a single whole-body exposure to 4 Gy gamma rays (cesium source). Findings indicated the chemical elevated enzyme activities of lactate dehydrogenase (LDH), alkaline phosphatase (AlP), and acid phosphatase (AP), as well as protein content and percentage of neutrophils in bronchoalveolar lavage fluid (BALF); the percentage of alveolar macrophages was sharply reduced. Radiation alone produced no substantial changes in the parameters investigated. Treatment with both agents combined was found to result in a synergistic rise of LDH, AlP, and AP activities and protein content in BALF. It was concluded that the BALF biochemical markers used are reliable indicators for identifying the type of combined effect produced in the lungs by chemical agents and ionizing radiation. PMID- 8243397 TI - Environmental pollutants in relation to complications of pregnancy. AB - Certain complications of pregnancy, e.g., threatened spontaneous abortion, toxemia, emesis, and anemia, were studied in pregnant women living in industrial areas contaminated by smelters and the petrochemical industry. Exposure to lead or aromatic hydrocarbons was assessed in parallel by the determination of these agents or their metabolites in blood and urine. Comparison of respective exposure levels was made between women with normal pregnancies and those with complications. Significantly higher levels of lead in blood and increased excretion of the metabolic products of organic solvents were found in women with complicated pregnancies compared to those with normal pregnancies. Threatened spontaneous abortion, toxemia, and anemia were associated with higher lead exposure in the vicinity of smelters. In these patients, evidence of disturbances of blood glutathione equilibrium and increased lipid peroxidation were found indicating a decreased ability to compensate for the effects of exposure. Styrene exposure in a petrochemical industrial area was associated mainly with late toxemia and nephropathy. Patients with these complications also had a tendency to elevated exposure to other aromatic hydrocarbons. It is suggested that complications of pregnancy may be induced by environmental agents at levels lower than those that result in pregnancy loss or preterm birth. PMID- 8243398 TI - Transformation of BALB/c-3T3 cells: I. Investigation of experimental parameters that influence detection of spontaneous transformation. AB - The frequency of spontaneous morphological transformation is an important variable in measuring chemical-induced transformation in BALB/c-3T3 clone A-31-1 13 cell cultures. Data from 110 experiments, which included benzo[a]pyrene control groups and other chemical treatment groups, were analyzed for factors that influenced spontaneous transformation. Spontaneous transformants demonstrated a continuum of morphological variants (type I, II, and III foci) that fit a normal distribution if converted to log10. The magnitude of transformation depended on the ampule of cryopreserved cells and the serum lot. Although the average frequency was approximately 0.71 x 10(-6) (type III foci/cell that survived and proliferated to confluence), the absolute number of foci/vessel increased in proportion to the surface area of the culture vessel. Thus, the frequency of spontaneous transformation was directly related to the cumulative number of mitoses that occurred in forming the contact-inhibited monolayer. These data are consistent with a hypothesis that spontaneous transformation in BALB/c-3T3 cells is a mutational event or some other single step phenomenon. PMID- 8243399 TI - Transformation of BALB/c-3T3 cells: II. Investigation of experimental parameters that influence detection of benzo[a]pyrene-induced transformation. AB - Benzo[a]pyrene (BaP) induced significant morphological transformation of clone A31-1-13 BALB/c-3T3 cells without exogenous activation. Therefore, BaP was selected as a model to determine the internal consistency of detection of chemical-induced transformation. BaP induced a continuum of type I-III foci of different sizes, and the ratio of type I-III to type III foci/vessel was usually about 2-fold. The major finding was that BaP induced highly significant transformation responses, and the magnitude of these responses were inversely correlated with the cytotoxicity of the treatment doses. Thus, the induction of BaP-induced transformation behaved as though it was caused by a mutational event. Variability among responses were shown to depend on the serum lot and the cryopreserved ampule of cells. In addition, experiments with low spontaneous transformation responses had an impaired ability to detect BaP; however, experiments with high or normal spontaneous responses had a normal ability to detect BaP. Because the expression of BaP-induced transformation depended on both the cytotoxicity of the treatment and the cumulative number of mitoses, the frequency of BaP-induced transformation should be reported as the number of foci/vessel, but not expressed as the number of foci/viable cell surviving the chemical treatment. These conclusions are important because the same 110 experiments described in this report were also used to evaluate the transformation responses of many different carcinogenic and noncarcinogenic chemicals. These data are being reported separately. PMID- 8243400 TI - Transformation of BALB/c-3T3 cells: III. Development of a co-culture clonal survival assay for quantification of chemical cytotoxicity in high-density cell cultures. AB - A co-culture clonal survival assay was developed to measure the cytotoxicity of test chemical treatments to BALB/c-3T3 cells because the standard clonal survival assay using 200 wild type (WT) cells frequently overestimates chemical cytotoxicity when compared with identical treatment doses in high-density cultures. The assay used co-cultures of 3.2 x 10(4) WT cells, the same seeding density used in the transformation assay, and 200 ouabain resistant (OUAr) cells. After a 48-hr test chemical treatment, co-cultured cells were fed with culture medium containing 4 mM ouabain. The test chemical was cytotoxic to an equal percentage of WT and OUAr cells. Ouabain treatments killed the remaining WT cells. Thus, OUAr cells surviving the test chemical treatment measured the relative cloning efficiency (RCE) of all treated cells in the high-density cell co-culture. The co-culture assay succeeded because metabolic cooperation at the OUAr locus was not detected in BALB/c-3T3 cells. Five chemicals induced comparable cytotoxic responses in both assays, including actinomycin D, 5-bromo 2'-deoxyuridine, N'-methyl-N-nitro-N'-nitrosoguanidine, dimethyl sulfoxide and sodium chloride. In contrast, chemical cytotoxic responses detected in the standard and co-culture assays differed by > or = 10-fold for 11-aminoundecanoic acid, benzo[a]pyrene, cytosine arabinoside, and 3-methyl-cholanthrene and differed by > 2-fold for 2-acetylaminofluorene and dimethylnitrosamine. Detection of 11-aminoundecanoic acid-induced transformation was shown to be dependent on selecting treatment doses from the co-culture assay data. Thus, this method permits more accurate assessment of both chemical-induced cytotoxicity and transformation. PMID- 8243401 TI - Transformation of BALB/c-3T3 cells: IV. Rank-ordered potency of 24 chemical responses detected in a sensitive new assay procedure. AB - This report introduces an improved method of detecting chemical-induced morphological transformation of A-31-1-13 BALB/c-3T3 cells. The new procedure uses an increased target cell population to assess chemical-induced damage by increasing the initial seeding density and by delaying the initiation time of chemical treatment. Furthermore, a newly developed co-culture clonal survival assay was used to select chemical doses for the transformation assay. This assay measured the relative cloning efficiency (RCE) of chemical treatments in high density cell cultures. In addition, transformation assay sensitivity was enhanced through the use of improved methods to solubilize many chemicals. From a group of 24 chemicals tested in at least two trials, clear evidence of chemical-induced transformation was detected for 12 chemicals (aphidicolin, barium chloride-2H2O, 5-bromo-2'-deoxyuridine, C.I. direct blue 15, trans-cinnamaldehyde, cytosine arabinoside, diphenylnitrosamine, manganese sulfate-H2O, 2-mercaptobenzimidazole, mezerein, riddelliine, and 2,6-xylidine); 2 chemicals had equivocal activity [C.I. direct blue 218 and mono(2-ethylhexyl)phthalate], 9 chemicals were inactive [carisoprodol, chloramphenicol sodium succinate, 4-chloro-2-nitroaniline, C.I. acid red 114, isobutyraldehyde, mono(2-ethylhexyl)adipate, sodium fluoride, and 12-O-tetradecanoylphorbol-13-acetate), and 1 chemical had an indeterminate response (2,6-dinitrotoluene). All positive responses were detected in the absence of an exogenous activation system and exhibited significant activity at two or more consecutive doses. This report also presents a mathematical method that uses t-statistics for rank-ordering the potency of chemical-induced transformation responses. This model detects sensitivity differences in experiments used to evaluate chemical-induced transformation. Furthermore, it provides a method to estimate a chemical's transformation response in terms of the historical behavior of the assay, as well as its future activity. The most active of the 24 chemicals was mezerein, and the least active chemical was diphenylnitrosamine. PMID- 8243402 TI - Transplacental transfer of genotoxins and transplacental carcinogenesis. AB - A number of chemical compounds induce cancer in the offspring of animals treated with these compounds. The fetus is sensitive to the toxic and teratogenic effects of chemicals in the early embryonic stages, whereas it is sensitive to carcinogenic effects during late fetal stages. Carcinogens may be direct acting or may require metabolic oxidation such as those in tobacco smoke. Activation can occur in utero. Animal experiments indicate that tumors can be initiated in utero, commonly by activation of cellular proto-oncogenes, and that promotion can occur after birth by postnatal treatment with tumor promoters. This may have important implications for humans. The initial peak of cancer incidence during the first 5 years of life may be due to prenatal exposure of either parent to mutagens, but the role of paternal exposure in relation to childhood cancer is controversial. There is an increased risk of cancer in children whose fathers work in heavy industry or whose mothers work in medical or dental services. The exact etiological agents have not been unequivocally identified. Information on human transplacental exposure to carcinogens and genotoxins is limited and based on measurement of maternal plasma concentrations or analysis of cord blood. Transplacental transfer of carcinogens in smoke and smoke-related damage to fetal tissue have been demonstrated. The mycotoxin aflatoxin B1 or its metabolites have been detected in cord blood, as have metabolites of pesticides and polychlorinated biphenyls. New biomarkers may provide important information on the transplacental transfer of genotoxic compounds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243403 TI - Transformation of BALB/c-3T3 cells: V. Transformation responses of 168 chemicals compared with mutagenicity in Salmonella and carcinogenicity in rodent bioassays. AB - This report describes the activities of 168 chemicals tested in a standard transformation assay using A-31-1-13 BALB/c-3T3 cells. The data set includes 84 carcinogens, 77 noncarcinogens, and 7 research chemicals. Carcinogens included 49 mutagens and 35 nonmutagens; noncarcinogens included 24 mutagens and 53 nonmutagens. The transformation assay did not use an exogenous activation system, thus, all chemical responses depended on the inherent target cell metabolic capacity where metabolic activation was required. The upper dose limit was 100 milli-osmolar because the assay could not discriminate active and inactive chemicals tested above this concentration. Certain physicochemical properties resulted in technical problems that affected chemical biological activity. For example, chemicals that reacted with plastic were usually nonmutagenic carcinogens. Similarly, chemicals that were insoluble in medium, or bound metals, were usually nonmutagenic and nontransforming. Multifactorial data analyses revealed that the transformation assay discriminated between nonmutagenic carcinogens and noncarcinogens; it detected 64% of the carcinogens and only 26% of the noncarcinogens. In contrast, the transformation assay detected most mutagenic chemicals, including 94% of the mutagenic carcinogens and 70% of the mutagenic noncarcinogens. Thus, transformation or Salmonella typuimurium mutagenicity assays could not discriminate mutagenic carcinogens from mutagenic noncarcinogens. Data analyses also revealed that mutagenic chemicals were more cytotoxic than nonmutagenic chemicals; 88% of the mutagens had an LD50 < 5 mM, whereas half of the nonmutagens had an LD50 > 5 mM. Binary data analyses of the same data set revealed that the transformation assay and rodent bioassay had a concordance of 71%, a sensitivity for carcinogens of 80.0%, and a specificity for detecting noncarcinogens of 60%. In contrast, Salmonella mutagenicity assays and rodent bioassays had a concordance of 63%, a sensitivity of 58%, and a specificity of 69%. The transformation assay complemented the Salmonella mutagenesis assay in the identification of nonmutagenic carcinogens; thus, the two assays had a combined 83% sensitivity for all carcinogens and a 75% specificity for nonmutagenic noncarcinogens. PMID- 8243404 TI - Environmental factors influencing growth and pubertal development. AB - Postnatal growth is based on hereditary signals and environmental factors in a complex regulatory network. Each factor must be in an optimal state for normal growth of the child. Fetal conditions may also have consequences on postnatal height. Intrauterine growth retardation can be recovered postnatally, although postnatal growth remains depressed in about one-third of cases. After birth, the environment may exert either a positive or negative effect on growth. In underdeveloped countries, malnutrition plays a major role in inhibiting the growth process. Children from families of higher socioeconomic classes are taller than their coevals in the lower socioeconomic groups. Urbanization also has a positive effect on growth. Better child care is supported by sufficient food supply, appropriate health and sanitation services, and a higher level of education. Over the last century, these factors have induced a taller stature and a more rapid maturity in Europe, North America, and Australia; a phenomenon which has been referred to as "the secular trend" in growth. Recently, a secular trend has also been reported in some developing countries. Although urbanization in general appears to be associated with better conditions of living, this is not the case in the slums of South America or in Africa where rural children are better off than children living in the poor cities. This paper describes in more detail the different hereditary and environmental factors that act during the fetal period and postnatally, and which play a role in human growth and pubertal development. PMID- 8243405 TI - Nurturing and breast-feeding: exposure to chemicals in breast milk. AB - All chemicals that are not normal constituents of human milk should be considered undesirable contaminants. In the present review, the following substances detected in human milk are considered: persistent organochlorine pesticides; polychlorinated biphenyls (PCB); polychlorinated dibenzodioxins (PCDD) and dibenzofurans (PCDF); polybrominated compounds; polycyclic aromatic hydrocarbons (PAH); trace elements; mycotoxins; nitrate, nitrite, nitrosamines; nicotine, caffeine, ethanol; and drugs. The levels of most of these substances found in human milk were within a range that would not constitute health hazards for breast-fed infants. For many of these, there is a comfortable safety margin. This applies also to organochlorine pesticides and PCB, particularly since, as a result of their discontinued use, the levels of these compounds have clearly declined in recent years. On the other hand, the aflatoxin burden mediated through breast milk, at least in certain tropical countries, appears to pose a definite health hazard. Detailed reference are given on the contamination of human milk with PCDD/PCDF which has to be considered as a matter of concern from the viewpoint of preventive public health. Although the low PCDD/PCDF levels found in the adipose tissue of infants indicate that there is no appreciable health risk emanating from these substances for breast-fed infants, appropriate measures to reduce the current rate of their emission into the environment have to be taken. PMID- 8243406 TI - Genetic anomalies in mammalian germ cells and their significance for human reproductive and developmental risk. AB - The induction of heritable mutations in germ cells represents a potential health concern. This paper highlights data from mouse germ-cell mutagenesis studies that have implications in the assessment of reproductive and developmental risks. The paper discusses the developmental and reproductive consequences of induced chromosomal damage (structural rearrangements and numerical anomalies) and describes environmental agents that have been shown to produce such anomalies. Additionally, factors that influence the yield of genetic damage are addressed. Studies showing that the various germ-cell stages vary in their susceptibility to the induction of genetic damage are summarized. Of the chemicals evaluated in the male mouse, most appear to have their predominant or strongest effect on post stem-cell stages. The differences between males and females in the susceptibility to mutagens is examined. Recent studies have shown that the female may be uniquely sensitive to certain mutagens. Finally, an important aspect of mutagenic risk is not only effects induced in developing germ cells but also the effects of environmental agents during the period from fertilization through the zygote and the two-cell embryo. Recent work in the mouse has demonstrated that exposure during these early developmental stages leads to high frequencies of external and visceral fetal malformations, as well as mid-to-late gestational death. PMID- 8243407 TI - Effects of environmental factors on perinatal outcome: neurological development in cases of intrauterine growth retardation and school performance of children perinatally exposed to ionizing radiation. AB - We performed two studies to investigate environmental factors in relation to neurological development in infants. The first, a field study, examined the elementary school performance of 929 children who were born from mothers exposed to the atomic bombing of Hiroshima, Japan, August 6, 1945. The most severe mental retardation was observed in the group exposed between 8 and 15 weeks following fertilization, and the second most severely damaged group was exposed between 16 and 25 weeks. The second, a clinical investigation, examined infants in the perinatal center who survived intrauterine growth retardation (IUGR). Those who survived with abnormal neurological development had a mean growth arrest corresponding to a uterine height of 27 weeks of gestation. This was at an earlier stage than those who survived with normal neurological development and had a mean growth arrest corresponding to 29-30 weeks of gestation. A smaller head circumference at birth was closely correlated with abnormal neurological sequelae. These results indicate that the brain development of the fetuses may have been affected by neurotoxic events similar to ionizing radiation. We emphasize the importance of avoiding neurotoxic stress to pregnant women when the fetus is in the critical period of neuronal development, before 27 weeks of gestational age. PMID- 8243408 TI - Stress and other environmental factors affecting fertility in men and women: overview. AB - To understand how environmental factors contribute to fertility or infertility in humans, it is first necessary to define environment. A view that will guide this review is that environment represents the "external milieu," analogous to the well-defined concept of "internal milieu" first introduced by Claude Bernard. Within this context, the environment provides both positive and adverse influences on reproductive health and development. Environmental factors can then be classified into categories such as physical, chemical, biological, behavioral, and socioeconomic. In many circumstances, multiple environmental factors may contribute to adversely modify human health. It has been suspected and in some cases demonstrated that stress can adversely affect reproductive function. Both animal and human data support this contention; however, the human data are clear in extreme situations (e.g., inmates of concentration camps) but less so under less drastic conditions. In recent years many advances have been made concerning the neurochemical mechanisms that mediate the effects of stress on reproductive functions and on the identification of "stress hormones" that may not only be involved in the stress response but also serve as biochemical markers to identify and correlate stress with different fertility parameters. Nutrition also plays an important role in infertility, and undernutrition or nutrition disorders are associated with stress in infertility. Environmental factors are often invoked as contributing to many cases of unexplained infertility. However, the direct causal relationship between those factors and the ensuing infertility of the couple are seldom well established and remain largely anecdotal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243409 TI - Regional and geographical variations in infertility: effects of environmental, cultural, and socioeconomic factors. AB - Fertility is affected by many different cultural, environmental, and socioeconomic factors, especially in developing countries where poverty and infections are commonplace. Environmental factors play a major role in infertility in Africa. One of the most important health problems in sub-Saharan Africa is the high rate of infertility and childlessness. The African society has a strong traditional heritage, and the study of the patterns of infertility in this part of the world would be incomplete without consideration of the sociocultural and environmental factors. The most cost-effective approach to solving the infertility problems in Africa is prevention and education. In Mexico, problems of reproductive health are associated with pregnancy in adolescents, sexually transmitted diseases and genitourinary neoplasms. Infertility affects 10% of couples, usually as a result of asymptomatic infection. Education, poverty, nutrition, and pollution are problems that must be tackled. The government has taken positive action in the State of Sao Paulo in Brazil, where gender discrimination is a major factor affecting women's health and reproductive outcomes. The implementation of new policies with adequate funding has resulted in marked improvements. PMID- 8243410 TI - Effects of the workplace on fertility and related reproductive outcomes. AB - This report reviews the recent literature on the adverse effects of occupational factors on fertility and related reproductive outcomes. Few studies fulfill the criteria of good study design because of small sample size, insensitive measures of effect, selection, recall, and observation bias, weak if any control of confounding factors, bad definition of exposure, inability to analyze a dose response relationship, and inadequate statistical analysis. The high prevalence of unsuccessful reproductive outcomes in the general population makes the design of human fertility studies difficult. Although a number of publications indicate that certain occupational factors and settings adversely affect both male and female fertility, it is virtually impossible to estimate the proportion of infertility due to occupational factors in the general population. The collected data suggest that the exposure to the following substances or occupational settings may affect a function of male genital system, leading to sperm abnormalities, hyperestrogenism, impotence, infertility, and/or increased spontaneous abortion rate in wives of exposed workers: alkylmercury, antimonide, anesthetic gases, boron, carbon disulfide, chlorodecone, chloroprene, some carbamates (carbaryl), diaminostilbene, 1,2-dibromo-3-chloropropane, ethylene glycol ethers, ethylene dibromide, inorganic lead, manganese, methyl chloride, organic solvents, synthetic estrogens and progestins, tetraethyllead, combined exposure to styrene and acetone, welding operations, and heat. The majority of reviewed papers on female fertility concerns the alterations of menstrual cycle and pregnancy complications rather than occupational exposure-induced female infertility. The literature supports the hypothesis that, in general, working women have a tendency of higher risk of unsuccessful reproductive outcomes, although the existing data are not sufficient. PMID- 8243412 TI - Urinary protein markers in pesticide applicators during a chlorinated hydrocarbon exposure. AB - A previous study of 16 pesticide applicators showed that N-acetylglucosaminidase (NAG) excretion during exposure to the soil nematocide, 1,3-dichloropropene (DCP), was correlated with DCP dose and the excretion of its major metabolite, N acetyl-S-(cis-3-chloroprop-2-enyl)-cysteine (3CNAC). Using the same urine samples, two other proteins were measured: low concentrations of albumin ("microalbuminuria," ALB) and retinol-binding protein (RBP). For each worker there were five consecutive urine collection intervals (6-8 hr each): overnight prior to exposure, morning work shift, afternoon work shift, evening after work, and overnight. Paired comparisons of each interval to baseline (first interval) for each protein excretion (creatinine corrected) did not reveal significant increases. However, amounts of renal proteins per interval, in contrast to creatinine-corrected concentrations, were more strongly correlated with each other and with 3CNAC concentrations within samples. Clear dichotomous differences in urinary protein excretion were seen for high versus low exposure (< or > 1.5 mg 3CNAC excretion/day) for NAG and RBP, but not for ALB. Based on earlier findings of a slight elevation of NAG and the minimal changes reported herein for RBP, the possible effects of short-term workplace exposure to DCP on renal protein excretion appear to be mild. PMID- 8243411 TI - Environment, human reproduction, menopause, and andropause. AB - As the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator is an integrator of hormonal, metabolic, and neural signals, it is not surprising that the function of the hypothalamogonadal axis is subject to the influence of a large array of environmental factors. Before puberty, the central nervous system (CNS) restrains the GnRH pulse generator. Undernutrition, low socioeconomic status, stress, and emotional deprivation, all delay puberty. During reproductive life, among peripheral factors that effect the reproductive system, stress plays an important role. Stress, via the release of corticotropin-releasing factor (CRF), eventually triggered by interleukin 1, inhibits GnRH release, resulting in hypogonadism. Effects of CRF are probably mediated by the opioid system. Food restriction and underweight (anorexia nervosa), obesity, smoking, and alcohol all have negative effects on the GnRH pulse generator and gonadal function. Age and diet are important determinants of fertility in both men and women. The age associated decrease in fertility in women has as a major determinant chromosomal abnormalities of the oocyte, with uterine factors playing a subsidiary role. Age at menopause, determined by ovarian oocyte depletion, is influenced by occupation, age at menarche, parity, age at last pregnancy, altitude, smoking, and use of oral contraceptives. Smoking, however, appears to be the major determinant. Premature menopause is most frequently attributable to mosaicism for Turner Syndrome, mumps ovaritis, and, above all, total hysterectomy, which has a prevalence of about 12-15% in women 50 years old. Premature ovarian failure with presence of immature follicles is most frequently caused by autoimmune diseases or is the consequence of irradiation or chemotherapy with alkylating cytostatics. Plasma estrogens have a physiological role in the prevention of osteoporosis. Obese women have osteoporosis less frequently than women who are not overweight. Early menopause, suppression of adrenal function (corticoids), and thyroid hormone treatment all increase the frequency of osteoporosis. Aging in men is accompanied by decreased Leydig cell and Sertoli cell function, which has a predominantly primary testicular origin, although changes also occur at the hypothalamopituitary level. Plasma testosterone levels, sperm production, and sperm quality decrease, but fertility, although declining, is preserved until senescence. Stress and disease states accelerate the decline on Leydig cell function. Many occupational noxious agents have a negative effect on fertility.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243413 TI - A radioactive tracer technique to determine in vivo the number of fibers in the lungs of rats following their administration by intratracheal instillation. AB - A radioactive tracer technique is described which enables the total number of fibers present in the lungs of rats to be estimated following administration of the fibers by intratracheal instillation. The glass fiber used in the study was irradiated with thermal neutrons to induce radioactive 24Na. A suspension of the radioactive fiber was administered to eight rats by intratracheal instillation and to two additional rats by intraesophageal instillation. The 24Na radioactivity in the rats was counted in vivo at 24 and 48 hr after administration, after which they were killed. The amounts of fiber in the lungs, in the gastrointestinal tracts, and excreted in feces were estimated radiometrically. On average 93% of the administered fiber was accounted for. The lungs were digested with sodium hypochlorite solution and aliquots of the resulting digest filtered through membrane filters which were clear for examination by phase-contrast optical microscopy (PCOM). The numbers of fibers in the lungs, estimated by PCOM, were well correlated with the in vivo counting rates at 48 hr, indicating that the latter can be used to provide an accurate index of the number of fibers retained in the lung at that time. PMID- 8243414 TI - Pulmonary response, in vivo, to silicon carbide whiskers. AB - Fischer rats were exposed to silicon carbide whiskers (SiCW), boron carbide whiskers (BCW), silicon carbide platelets (SiCP), or crocidolite asbestos separately administered by intratracheal instillation. SiCW proved to be the most toxic material within the test group. Dramatic increases in alveolar macrophage populations within 1 week of exposure to SiCW persisted for at least 28 days, evidence of the chronic inflammation observed in necropsies during the first months of the study. The most common finding in histological preparations of tissues taken from animals 18 months after exposure to SiCW was a high incidence (frequency > 0.85) of multiple pulmonary granulomas which occasionally occluded airways. Lesions associated with crocidolite were similar to those found with SiCW. Equivalent treatment with BCW and SiCP produced no significant histological changes within 18 months of exposure. PMID- 8243415 TI - Physiologically based modeling of 2-butoxyethanol disposition in rats following different routes of exposure. AB - 2-Butoxyethanol (BE) is widely used as a solvent in coatings and other consumer products and has shown hematotoxicity in laboratory animals. To provide a physiological basis for extrapolating toxicokinetic data observed in rats to humans, a blood flow rate-limited, physiologically based pharmacokinetic model was developed to describe the distribution and metabolism of BE in rats following drinking water, dermal, and inhalation exposures. The major urinary metabolite, butoxyacetic acid, represented 45 to 60% of the absorbed dose in all three routes of exposure. Other identified urinary metabolites in our studies included ethylene glycol and BE-glucuronide. A model formulation of the possible metabolic pathways based on the experimental data was proposed. The amounts of individual urinary metabolites were used to develop the model. Metabolic constants were estimated by fitting the data within the constraints of in vitro measurements. The model explained the change of profiles of urinary metabolites in different exposure routes by taking into account the differences in absorption rate and by incorporating a minor pathway for metabolism by skin. Sensitivity analysis showed that metabolic constants and blood flow rate to liver had a relatively larger influence on the production of urinary metabolites than the organ volume or the partition coefficient for BE. PMID- 8243416 TI - Occupational influences relative to the burnout phenomenon among Japanese nursery school teachers. AB - To identify and evaluate recent working conditions and job content of nursery school teachers in Japan, as well as the prevalence of the burnout phenomenon and the occupational influences responsible for it, a questionnaire survey was carried out. The subjects consisted of 719 nursery school teachers and 204 municipal clerical workers as the control group. Working conditions and workload burdens were more severe among nursery school teachers than those of the clerical workers. The burnout phenomenon among the nursery school teachers was characterized by emotional exhaustion. Moreover, the rate ratio and multivariate analyses indicated that a great variety of occupational factors, not only interpersonal relationships, but also the general working conditions and specific physical or mental workloads, influenced the burnout phenomenon as well. Therefore, in examining measures dealing with the burnout phenomenon among nursery school teachers, it is important to evaluate the occupational factors systematically and comprehensively. PMID- 8243417 TI - Comparative study of modification and degradation of neurofilament proteins in rats subchronically treated with allyl chloride, acrylamide, or 2,5-hexanedione. AB - Allyl chloride (ALL), acrylamide (ACR), and 2,5-hexanedione (2,5-HD) are all industrial neurotoxicants and known to produce accumulation of neurofilament (NF) proteins in both the central and peripheral nervous systems. To clarify whether any common mechanisms underlie these neurofilamentous axonopathies, the ability of ALL, ACR, and 2,5-HD to cross-link the NFs and the effects on NF degradation by Ca(2+)-activated neural protease were investigated in spinal cords from rats subchronically treated with these chemicals. Sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by immunoblot analysis revealed the appearance of high-molecular-weight species of NF triplets immunoreactive to each anti-68K, anti-160K, and anti-200K NF antibody in the 2,5-HD-treated rats, whereas it was not found in those treated with ALL or ACR. A time course study on the degradation of NF proteins conducted by the co-incubation with Ca2+ showed degradation resistance in all three NF subunits from animals treated with 2,5-HD, while no significant alterations in the rate of NF degradation were observed in the ALL- or ACR-treated group. The present results suggest that neurofilament filled axonopathy induced by ALL or ACR and axonopathy induced by 2,5-HD may not share a common mechanism, though the initial step for the pathogenesis of this chemically induced neurotoxicity is not fully understood at present. PMID- 8243418 TI - Profile of subjective complaints and activities of daily living among current patients with Minamata disease after 3 decades. AB - We surveyed 1144 current patients with Minamata disease (MD) aged 40 or over in the Minamata area and the same number of neighbor controls matched with age and sex by questionnaire interview with regard to subjective complaints and activities of daily living (ADL). From analysis of subjective complaints, it was found that MD patients had significantly higher rates of all complaints than controls (P < 0.05). Multivariate analysis showed that subjective complaints in controls were clearly separated into the following two categories: sensory disturbances and movement nerve disturbances, but all complaints in MD patients formed one cluster. Such variation seemed to be due to methylmercury exposure to the central nervous system. ADL analysis revealed that the difference in the ADL disability between MD patients and controls significantly increased with age (P < 0.05) and that ADL disability in MD patients was aggravated by aging. PMID- 8243419 TI - Carbon disulfide and the central nervous system: a 15-year neurobehavioral surveillance of an exposed population. AB - Carbon disulfide-induced neurobehavioral effects are well known and do not need further evidence. Carbon disulfide vasculopathy and the syndromic complex resulting in depression, loss of memory and concentration, and behavior disturbances have been widely demonstrated. Less known is the evolution of the symptomatology when the environmental conditions are consistently improved, that is, the reversibility or the progression of the dysfunctions observed. This paper reports on a neurobehavioral follow-up in a viscose rayon factory carried out, in intervals, from 1974 to 1990. Several successive improvements were implemented in the plant through the years, until finally, the most radical changes were made at the end of the Seventies and these resulted in exposure levels far below the current Threshold Limit Values. A total of 493 subjects were examined and some of them were reexamined up to six times. The last examination was completed in September, 1990. In this paper, studies by our group over the 15 years of monitoring are discussed. The results show that the general mental state, as measured by neurobehavioral methods, reflects past and current exposure. This point was explored by dividing the subjects into six groups on the basis of their length of exposure and year of examination and by comparing their performances. The results show that even exposure to levels of carbon disulfide not exceeding 8 mg/m3 may induce absentmindedness and difficulties in perceptive abilities. PMID- 8243420 TI - Effects of methylmercury on protein kinase A and protein kinase C in the mouse brain. AB - The effects of methylmercury administration on adenosine 3',5'-cyclic monophosphate (cAMP)-dependent protein kinase (protein kinase A) and protein kinase C were investigated by determining their second messenger bindings ([3H]cAMP binding for protein kinase A and [3H]PDBu for protein kinase C) and enzymatic activities in the brains of methylmercury-treated mice. After single administrations of methylmercury (10 mgHg/kg, sc), no neurological symptoms were observed, while the mercury concentration in the brain reached 5.6 ppm. Neither second messenger bindings nor enzymatic activities of either protein kinase displayed significant changes. When methylmercury was administered repeatedly (10 mg Hg/kg x 5), the mercury concentration was 11.7 ppm and the enzymatic activity of protein kinase C was reduced to 75% of the control level without significant change in [3H]PDBu binding. Significant change has not been observed in either [3H]cAMP binding or enzymatic activity of protein kinase A. The reduction of enzymatic activity of protein kinase C was reversed by the simultaneous administration of selenite (0.5 mgSe/kg x 5). However, the fact that selenite administration alone displayed not a significant but about a 20% increase in [3H]PDBu binding suggested that selenite itself could affect the level of protein kinase C despite having no apparent effects on protein kinase C in vitro. Further investigation is necessary to assess whether protein kinase C is involved in the detoxication mechanism of selenite with respect to methylmercury. Since the mercury concentration in the brain was higher than the IC50s for both protein kinase A and protein kinase C observed in vitro even after single administration, methylmercury might inhibit both protein kinases, which might impair intracellular signal transduction. This might in part conceal the symptoms during the early stages of methylmercury toxicity. PMID- 8243421 TI - Neurochemical approaches to developing biochemical markers of neurotoxicity: review of current status and evaluation of future prospects. PMID- 8243422 TI - Neurotoxic effects of 2,5-hexanedione on rapidly growing unmyelinated peripheral nerve axons of a rat fetus: dose-effect relationship. AB - To investigate the potential neurotoxicity of 2,5-hexanedione (2,5HD) on developing axons we examined peripheral nerves of rat fetuses. Pregnant female Sprague-Dawley rats were injected subcutaneously with 680 mg/kg of 2,5HD once a day from Day 12 of gestation (GD12) to GD16 in one group and with 340 mg/kg of 2,5HD once a day from GD12 to GD20 in the other group. On GD20 live fetuses were removed from the uteri and their sciatic nerves were examined morphologically. By electron microscopical observations, affected nerves revealed axons which were aggregated and fused together, but there were no axons aggregated with neurofilaments. The diameter distributions of axons revealed an increase in the number of small-size axons in the nerves of the 340 mg/kg group and showed an enlargement of part of the axons in the nerves of the 680 mg/kg group, suggested by the appearance of a second peak at a diameter larger than that of the first peak. PMID- 8243423 TI - Neurobehavioral changes among workers in some chemical industries in Egypt. AB - This study investigates the long-term neuropsychiatric manifestations of single or combined chemicals: manganese; zinc phosphide; lead, mercury, and TNT; and pesticides among exposed industrial workers. We found that 75% of the exposed subjects as a whole and 50% of those exposed to each of Zinc phosphide and pesticides presented with more than one neuropsychiatric symptoms or signs. The main signs were mask faces, hyporeflexia, hyperreflexia, peripheral neuropathy, static tremors, radiculopathy, muscle weakness, mental changes, fasciculations and tremors, wasting, hypotonia, abnormal deep reflexes, and sensory hyposthesia. Neurological manifestations were confirmed by electromyography and their severity was related to the duration of exposure and confirmed as well by electroencephalography. These results are discussed and their implications high lighted. PMID- 8243424 TI - Neuropsychological assessment of organic solvent effects in South Africa: test selection, adaptation, scoring, and validation issues. AB - The neurobehavioral effects of organic solvent exposure were assessed in 68 South African workers in a paint manufacturing plant in which the maximum current combined solvent level in workplace air was 0.72 of the Threshold Limit Value. A 17-test battery was assembled from the WHO-NCTB, NES 2, and the University of South Africa Neuropsychological Assessment Procedure. Extensive modifications were made to some tests in the battery, using a guided learning rationale derived from the cross-cultural literature. In light of test correlations with demographic variables (age, education, and alcohol consumption), the construct and likely predictive validity of the tests in the battery are reviewed, and promising procedures identified. Finally, it is noted that in developing countries, the results of neuropsychological tests that have been well standardized in the West may be misleading unless the underlying validity issues that arise when a test developed in one culture is applied to another have been addressed. PMID- 8243425 TI - Calcium intakes in individuals on diets for the management of cows' milk allergy: a case control study. AB - Three hundred and twenty-three individuals with self-reported food allergy were recruited by media advertisements. Questionnaire information was collected on all respondents. Chocolate (57%), milk (47%), wheat (36%) and food additives (35%) were the most frequently implicated foods. The most frequently reported symptoms were itching (43%), skin rash (43%) and tiredness (43%). Food avoidance was the most common form of food allergy diagnosis (33%) with only 8% of respondents reporting food challenge in food allergy diagnosis. Self-diagnosis was reported by 34% of respondents with 29% and 24% reporting diagnosis by a general practitioner or a homeopath, respectively. Twenty-four per cent of respondents 'always' avoided and a further 57% 'nearly always' avoided the implicated food(s). A group of 38 adults with self-reported 'milk allergy' was selected for further study. Dietary assessments, using the dietary history method, were carried out on this subgroup and on age-, sex- and occupation-matched controls. The results of the dietary assessments revealed that the 'milk allergy' group had significantly higher intakes of fibre, beta-carotene, vitamin C, vitamin E, iron and folic acid (P < 0.05) and significantly lower intakes of calcium (P < 0.002) in comparison to controls. Eighteen per cent of the group used milk alternatives. Thirty-four per cent of the 'milk allergy' group took calcium-containing supplements. Even after calcium supplementation, the mean calcium intake of those who completely avoided milk was unacceptably low (441 mg/d). PMID- 8243426 TI - Osteoporosis and bone metabolic parameters in dependence upon calcium intake through milk and milk products. AB - The bone mineral content of young adults as well as of osteoporotic patients and age-matched controls without bone disease was measured by single-photon absorptiometry. A retrospective nutrition survey was additionally made to study the relationship between bone mineral content and calcium intake in different periods of life. The bone mineral content and bone mineral density of young adults is directly related to the calcium intake through milk and dairy products. The osteoporotics had a significantly lower bone mineral content than the controls. Calcium intake through milk and milk products in childhood and adolescence had been significantly lower in the patients than in the controls, whereas in the later periods of life (20-30 years prior to the study and at the time of the study) there were no significant differences between the calcium intakes of the two groups. It was also found that an adequate intake of calcium protected against increased bone resorption, as evidenced in particular by the reduced levels of serum osteocalcin, a parameter of bone turnover. In conclusion it can be stated that the data support the hypothesis that adequate calcium intake through milk and milk products in childhood and adolescence is a decisive marker for obtaining a maximum bone mass (peak adult bone mass) and for the prevention of osteoporosis. Furthermore, it can be stated that increased calcium intake in the later years may not reduce the accelerated risk of osteoporosis resulting from inadequate calcium intake during childhood and adolescence. PMID- 8243427 TI - Effects of two low-fat diets, high and low in polyunsaturated fatty acids, on plasma lipid peroxides and serum vitamin E levels in free-living hypercholesterolaemic men. AB - Diet enriched with polyunsaturated fat may increase the susceptibility of LDL to oxidation. Therefore the effects of two low-fat diets on plasma lipid peroxides in free-living mildly hypercholesterolaemic men (n = 37) were investigated in a randomized single-blind 28-week study. Composition of the diets were (1) American Heart Association (AHA) type 32/10:8:8 (indicating percentages of energy from total fat/saturated fat:monoenes:polyenes in actual diet); (2) low-fat 30/12:8:3. The subjects kept 3-day dietary records five times during the study to estimate the intake of nutrients. Plasma lipid peroxides were measured photometrically as the thiobarbituric-acid reactive substances (TBARS). Levels of serum vitamin E during the study were also determined. Mean change (+/- SD) in serum low density lipoprotein (LDL) cholesterol was similar in both groups (-0.32 +/- 0.76 vs -0.32 +/- 0.87 mmol/l) (AHA type vs low-fat). Level of TBARS decreased (P < 0.05) during the AHA type diet (-8.4 +/- 37.1%) (mean +/- SD) and increased (P = 0.228) during the low-fat diet (+8.7 +/- 27.0%) from 0 to 6 months. The mean intake of total active tocopherols was greater (14.7 +/- 3.7 mg) during the AHA type diet compared to the low-fat diet (7.8 +/- 2.1 mg). Serum vitamin E to LDL cholesterol ratio increased from 8.9 +/- 2.9 to 9.6 +/- 2.4 nmol/mmol (0 vs 6 months) (P = 0.07) during the AHA type diet and from 8.6 +/- 2.6 to 9.3 +/- 2.4 nmol/mmol (P = 0.159) during the low-fat diet.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243428 TI - The effect of a natural high-fibre diet on faecal and biliary bile acids, faecal pH and whole-gut transit time in man. A controlled study. AB - Dietary fibre possibly protects against colonic cancer by effects on bile acid metabolism. We investigated the effect of a natural high-fibre diet on secondary bile acid formation. Twelve healthy subjects on an habitual low-fibre diet (for 4 weeks) consumed a high-fibre menu for 10 weeks (experimental group). A control group of 10 subjects consumed their regular high-fibre diet during this period. Faecal and biliary acid composition, faecal weight, faecal pH and gut transit time were studied before and after 6 and 10 weeks of fibre addition. Changes in the experimental group were compared to changes in the control group. The concentration, but not the excretion, of the secondary faecal bile acids was reduced in the experimental group. Faecal weight increased, faecal pH dropped and gut transit time was not altered. The biliary deoxycholic acid content decreased and the cholic acid content increased after 6 weeks, but returned to baseline values after 10 weeks of fibre addition. This study shows that a natural high fibre diet lowers secondary faecal bile acid concentration through an increase in stool weight. The 7 alpha-dehydroxylation of primary bile acids is probably not or only transiently inhibited. PMID- 8243429 TI - Postprandial de novo lipogenesis in alcoholic liver cirrhosis: relationship with fuel homeostasis and nutritional status. AB - The prevalence and nutritional consequences of postprandial de novo lipogenesis were evaluated in 24 stable alcoholic cirrhotic patients. Energy expenditure and the rates of nutrient oxidation were assessed by indirect calorimetry after an overnight fast and 2 h after a standard meal supplying 15 kcal/kg (63 kJ/kg) of body weight. Postprandially, net lipogenesis occurred in 16 patients (group L+) as shown by a respiratory quotient clearly above 1.00 (P < 0.01). The rate of lipid oxidation remained positive in 8 patients (group L-). The main mechanism involved in this metabolic pathway appeared to be a sharp postprandial hyperinsulinaemia. When compared to group L+, patient group L- showed an impaired thermic effect of food (P < 0.05), a lower rate of glucose oxidation (P < 0.05) and a mild hyperketonaemia (P < 0.05) at fasting levels. Muscular and fat masses were lower (respectively P < 0.05 and P = 0.05) and the severity of the disease as assessed by the Child-Pugh classification was more pronounced in this group (P < 0.02). The occurrence of postprandial lipogenesis in stable cirrhotic patients is related to better nutritional status. Such a metabolic pathway may explain the nutritional heterogeneity of cirrhotics and is likely to have an effect on the benefits of refeeding. PMID- 8243430 TI - Acute effects of graded alcohol intake on glucose, insulin and free fatty acid levels in non-insulin-dependent diabetic subjects. AB - Alcohol-induced hypoglycaemia is a well-known phenomenon in insulin-treated diabetic subjects. Less attention has been paid to the impact of alcohol on blood glucose and insulin responses in non-insulin dependent diabetic subjects. The aim of this study was to investigate the acute metabolic effects of different alcohol contents added to a non-alcohol beer in 10 non-insulin-dependent diabetes mellitus (NIDDM) subjects. The patients received 500 ml non-alcohol beer with an alcohol percentage (v/v) of 0 (A), 2.7 (B), and 5.4 (C), implying identical contents of ingredients except for alcohol. Blood glucose (mean +/- SE) responses were similar in the three situations (395 +/- 59, 365 +/- 86 and 261 +/- 26 mmol/l x 240 min). In contrast, the incremental insulin response areas increased dose dependently to alcohol (5430 +/- 1158, 9336 +/- 2172 and 12336 +/- 2922 pmol/l x 240 min) and showed a linear correlation (r = 0.39; P < 0.03). The average suppression of serum free fatty acid was similar in the three situations (72.4 +/- 4.4%, 76.3 +/- 6.0% and 68.2 +/- 6.3%). In conclusion, intake of small amounts of alcohol does not acutely deteriorate the glycaemic control in NIDDM. The fact that alcohol results in a dose-related elevation in insulin levels with unaltered blood glucose and free fatty acid responses in NIDDM points to an aggravation of insulin resistance. PMID- 8243431 TI - Body mass index and work-disabling morbidity: results from a Bangladeshi case study. AB - A classification for chronic energy deficiency (CED) among adults has recently been proposed which utilizes body mass index (BMI). This paper reports on data collected in an urban slum in Bangladesh on the association between BMI and work disabling morbidity among adult men (n = 199). The mean BMI among the study men was 19.0 (SD = 2.9). 9% were classified as grade III CED, 15.6% grade II, 27% grade I, and 48.2% as 'normal'. There was a significant inverse association between BMI and work-disabling morbidity. Below a BMI of 16.0, 55% of men had lost one or more working days in the month prior to interview. This proportion dropped to 35% among those with a BMI between 16.0 and 17.0. Above a BMI 'threshold' of 17.0, the percentage of men incapacitated from work was similar in each BMI category. These results held whether accidents were included or excluded. Although the results are limited by the cross-sectional design of the study, the association supports the proposed classification of CED among adults. It is hoped that these results stimulate further research in this area. PMID- 8243432 TI - Elevated glutathione S-transferase activity in erythrocytes from malnourished children. AB - Glutathione S-transferases (GSTs) are principally involved in detoxification. These enzymes can be induced by an increased flux of substrate, such as occurs during pro-oxidative stress or antioxidant deficiency. We tested the hypothesis that the postulated oxidative stress in severe malnutrition would result in induction of GSTs in erythrocytes. Erythrocyte GST activity towards 1-chloro-2,4 dinitrobenzene (CDNB) was measured in 271 malnourished children (22 undernourished; 92 marasmic; 82 kwashiorkor; 75 marasmic-kwashiorkor) and 48 healthy children. GST activity in the malnourished children was significantly higher than the control group (P < 0.01). The GST activity in the four classes of malnutrition did not differ. There was a weak relationship between GST activity and the height deficit, but not with the weight deficit, or the clinical features displayed by the children. The 11 children that died had a higher value than the survivors. There was no change in GST with anthropometric recovery. We conclude that erythrocyte GST has been induced in children with malnutrition. Induction of erythrocyte GST may be the result of exposure of the children to oxidative stress during the months prior to their presentation with severe malnutrition. PMID- 8243433 TI - Dietary assessment in Asian children--a comparison of the weighed inventory and diet history methods. AB - A cross-sectional study of the dietary intake of Indo-Asian children, in Sheffield, aged 4-40 months, was carried out during a 1 year period. Weighed dietary inventories (facilitated by use of the PETRA electronic recording scale) and diet histories were completed, and analysis was carried out on the records of 117 children. This number is equivalent to a quarter of the estimated target population and 53% of those approached. The diet history method gave higher estimates of the dietary intake of most nutrients than the weighed inventory method (Wilcoxon's rank sum method), the differences represented 7%, 9%, 3%, 9% and 6% of the mean intakes of energy, protein, fat, iron and vitamin C respectively, and were significant for energy, protein and iron. Analysed by age group, differences were significant for energy at 12 to < 18 months; for iron at 6 to < 12 months and 12 to < 18 months; and for vitamin C at < 6 months. There were no significant differences between the estimates of standardised energy or protein intakes, nor in the estimates, by the two methods, of total energy (%) derived from protein or fat. Mean energy intake per kg was equivalent to basal metabolic rate (BMR)* 1.49 by weighed inventory and BMR*1.58 by diet history. Energy intake was expressed as a ratio of age-adjusted 'reference' energy expenditure [A.M. Prentice et al. (1988) The Lancet 2, 1066-1069], the modal ratio energy intake/energy expenditure was 0.75 to < 1.0 for both weighed inventory and diet history methods, suggesting that the observed levels of energy intake were within a reasonable range. PMID- 8243434 TI - Enzyme-linked immunosorbent assays for gliadin and ovalbumin and their application in normal subjects. AB - Two quick, sensitive and direct sandwich enzyme-linked immunosorbent assays (ELISAs) for the measurement of gliadin and ovalbumin in serum have been developed. The ovalbumin assay has a sensitivity of 0.14 ng/ml (2 SD from zero). Cross-reactivities with ovomucoid and conalbumin were 58% and 0.2%, respectively. No cross-reactivity was observed with bovine serum albumin (BSA), human serum albumin (HSA), gliadin and beta-lactoglobulin. The gliadin assay did not cross react with BSA, HSA, ovalbumin and beta-lactoglobulin and had a sensitivity of 7 pg/ml. In an acute feeding study, of 6 h duration, serum ovalbumin levels were measured in five non-food-allergic subjects who had been given raw eggs (5 g ovalbumin) after an overnight fast. Following a period of at least 1 week the same subjects were given 132 g wholemeal bread (5 g gliadin) after an overnight fast, and serum gliadin levels were measured. After the consumption of raw eggs ovalbumin levels peaked between 210 and 300 min with maximum serum concentration between 0.52 and 31.08 ng/ml. After the consumption of bread, gliadin levels reached a maximum between 180 and 300 min, reaching peak levels between 1.12 and 12.2 ng/ml. We conclude that levels of detected ovalbumin were higher than those for gliadin in this group of individuals. PMID- 8243435 TI - Cerebellar neuronal activity correlates with spike and wave EEG patterns in the rat. AB - In this study we investigated the involvement of the cerebellum in high voltage spike-and-wave spindles, a rodent model of petit mal epilepsy. High voltage spindles, recorded epidurally from the sensorimotor neocortex, were correlated with single or multiple unit activity in the cerebellar cortex and deep cerebellar nuclei. The majority of neurons or neuronal groups in the cerebellum (77.9%) fired rhythmically and phase-locked with the high voltage spindles, either during the spike (43.2%; n = 41) or during the wave (34.7%; n = 33) component of the high voltage spindle. Tremor of the head and neck musculature, recorded with an accelero-meter, occurred during the high voltage spindle in approximately half of the rats. When present, rhythmic movement occurred predominantly during the wave phase of the high voltage spindle. The remaining half of the rats did not show tremor during high voltage spindles but, nevertheless, had cerebellar units that burst during the spike or wave phase of the high voltage spindle. These latter results demonstrate that phase-locked bursting of cerebellar units during high voltage spindle is independent of rhythmic movement. The findings suggest that rhythmic output from the cerebellum may contribute to the maintenance of generalized petit mal seizures. PMID- 8243436 TI - Motor and electrocorticographic epileptic activity induced by 3-mercaptopropionic acid in immature rats. AB - The convulsant action of 3-mercaptopropionic acid (3-MPA), a known inhibitor of glutamate decarboxylase activity, was studied in 7-, 12-, 18- and 25-day-old rats and in adult animals. 3-MPA elicited predominantly clonic, minimal seizures as well as generalized tonic-clonic (major) seizures at all developmental stages studied. The CD50 for major seizures did not change during development; CD50 for minimal seizures was significantly lower in 18-day-old rats than in older animals. Latency to the onset of seizures was shortest in 18-day-old rats and extremely long in 12- and, especially, in 7-day-old rats. This long latency might signify either changing molecular properties of glutamate decarboxylase during development or slow turnover of GABA at early postnatal stages. Electrocorticographic recordings demonstrated sharp EEG components in the frontal region as a first sign of 3-MPA action, and seizure patterns exhibited similar developmental changes as found with other seizure models (a decrease in duration of individual graphoelements and an increase in synchronization among various cortical regions). This indicates the primary importance of brain maturation in the expression of epileptic EEG phenomena. The correlation between EEG and motor phenomena was poor in the youngest animals and it ameliorated with age, but it never became perfectly coincidental. PMID- 8243437 TI - Cross-tolerance between carbamazepine and valproate on amygdala-kindled seizures. AB - Carbamazepine and valproate are two clinically used anticonvulsants which are also effective in the treatment of manic-depressive illness. Although the biochemical profiles of these drugs are markedly different, some mechanisms in common may be implied by their partially overlapping spectrum of therapeutic efficacy in seizure and affective disorders. Further evaluation of common biological targets of these agents was attempted by determining whether cross tolerance would occur to the anticonvulsant effects of carbamazepine and valproate on amygdala-kindled seizures. It had previously been shown that tolerance to carbamazepine's anticonvulsant effects on amygdala-kindled seizures occurs only with contingent drug administration, i.e., it occurs only when the drug is injected before the kindling stimulation, and not when the drug is given after the seizure. In the current studies, rats that were made tolerant to carbamazepine showed cross-tolerance to valproate. Kindled rats given carbamazepine after each seizure stimulation (i.e., non-tolerant controls) did not show tolerance to valproate's anticonvulsant effects, indicating that the cross-tolerance between carbamazepine and valproate was also contingent. The clinical implications and potential common biochemical target mechanisms of the cross-tolerance between carbamazepine and valproate deserve further investigation. PMID- 8243438 TI - Changes in the threshold of pentylenetetrazol-induced seizures in cats with a chronic granuloma in brain amygdala. AB - A parenchymal brain granuloma is the final consequence of a localized inflammatory reaction against an infectious agent, particularly parasites. In humans, these lesions are frequent causes of epilepsy. We studied the pattern of seizures induced by pentylenetetrazol (PTZ) in cats with a chronic granuloma induced 1 year earlier by injection of silicates into the brain amygdala. Compared with controls, the seizure threshold to myoclonic seizures was significantly decreased in cats with granuloma, while the threshold for tonic clonic seizures was not altered. Thus, the difference in PTZ doses between the initial and final components of the seizure became significantly enlarged (P < 0.001). Our results indicate that a granulomatous lesion in amygdala facilitates the generation and propagation of myoclonic seizures, whereas other components of PTZ-induced seizures are not affected. PMID- 8243439 TI - The effect of naloxone on cerebral blood flow and glucose metabolism in patients with complex partial seizures. AB - We used positron emission tomography with [15O]water and [18F]fluoro-2 deoxyglucose (FDG) to study the effect of naloxone on cerebral blood flow (CBF) and glucose metabolism (LCMRglc) in patients with complex partial seizures. There was no effect on glucose metabolism, but blood flow was reduced 7-12% 45-60 min after infusion of 1 mg/kg naloxone, as was the degree of lateral temporal CBF asymmetry in patients with > 10% baseline hypoperfusion. Endogenous opiates are involved in regulation of human CBF, and possibly in hypoperfusion in epileptic foci. Since [18F]FDG PET measures mean LCMRglc over the tracer uptake and scanning periods, it is inferior to [15O]H2O PET for detecting drug effects which may be time dependent. PMID- 8243440 TI - Valproate and palmitate binding to serum albumin in valproate-treated patients. Relation to obesity. AB - Binding of valproate and palmitate to serum albumin was studied in 29 valproate treated epileptic patients. The results were compared with similar observations in a reference group of 43 non valproate-treated individuals. The binding affinity for palmitate was decreased (P < 0.0001) resulting in increased availability of long-chain fatty acids (P = 0.008) due to competitive valproate binding in the valproate-treated patients. The findings support a hypothesis on the pathogenesis of obesity as a complication of valproate treatment of epilepsy. PMID- 8243441 TI - The initial development of a health-related quality of life model as an outcome measure in epilepsy. AB - Patients with refractory epilepsy, despite no fixed physical deficit, are often socially and psychologically handicapped. Currently available outcome measures for epilepsy do not adequately address these manifestations or their influence on well-being and quality of life. A patient-based health-related quality of life (HRQL) model for epilepsy including physical, social and psychological domains was constructed. It contains previously validated measures of anxiety, depression, happiness, overall mood, self-esteem, mastery, social satisfaction and general health and a specifically designed seizure severity scale with patient- and carer-based components. The psychometric properties of this model were evaluated in the context of the trial of a potential new antiepileptic drug. All the scales, except the Social Problems Questionnaire, have acceptable internal consistency (alpha 0.69-0.85) in this patient population. Construct validity is indicated by the ability of the scales to differentiate between groups of patients predicted to have different levels of psychosocial function. Treatment effects were detected by the patient (P = 0.017) and carers (P = 0.035) subscales of the seizure severity scale, the happiness (P = 0.003) and the mastery (P = 0.003) scales. Despite obvious deficiencies preliminary analyses are encouraging. This model provides a framework for investigating the complex interaction between the physical, social and psychological manifestations of epilepsy. The model has potential as an outcome measure for use in longitudinal studies and as a measure of disability for use in cross-sectional studies designed to compare quality of life in different populations of people with epilepsy. PMID- 8243442 TI - Measuring the impact of epilepsy: the development of a novel scale. AB - The impact of a chronic illness is experienced not only through its physical symptoms, but also as a result of its effect on psychosocial functioning. In the case of an illness such as epilepsy, where the physical manifestations are transient, the psychosocial consequences may, with time, come to be of greater concern. We have been involved in developing a quality of life model for epilepsy. As part of the refinement of the initial model, we have devised a novel scale to measure the impact of the condition on a number of different aspects of daily life. The scale was administered to 75 patients attending an epilepsy out patient clinic. Initial analysis of its psychometric properties is encouraging, although the inclusion of an item relating to employment reduced the scale's reliability. As a result, the wording of the existing item has been amended and an additional item has been incorporated. We hope the scale will be useful in investigations of treatment for epilepsy and of its psychosocial aspects. PMID- 8243443 TI - Disorders of puberty in boys. AB - Puberty is the last phase of the complex process of sexual maturation, a process by which an individual acquires reproductive competency. This article reviews the physiologic and physical changes of normal puberty and the causes of either delayed or precocious sexual development of boys. PMID- 8243444 TI - Menstrual disorders in adolescence. AB - Current concepts of normal and pathologic puberty have been reviewed because menstrual abnormalities are usually due to abnormal maturation of the reproductive endocrine system. Schema are presented for the initial evaluation of patients with amenorrhea or oligomenorrhea based upon whether they are hypoestrogenic or estrogenized. The role of the bone age in the differential diagnosis of delayed puberty is emphasized. The differential diagnosis and management of excessive genital bleeding and dysmenorrhea in adolescence are given. PMID- 8243445 TI - Acne, hirsutism, and alopecia in adolescent girls. Clinical expressions of androgen excess. AB - Hyperandrogenism must be considered in any girl with premature pubarche, unusual acne, hirsutism, or androgenetic alopecia. An association with menstrual irregularity or obesity should raise the index of suspicion. The most common causes of hyperandrogenism presenting in a teenage girl are functional ovarian hyperandrogenism, one manifestation of which is polycystic ovary syndrome, and functional adrenal hyperandrogenism, which usually seems to be due to an exaggeration of adrenarche. The plasma-free testosterone is a more sensitive indicator of hyperandrogenism than is the total testosterone concentration. The pattern of response of plasma free testosterone, DHEAS, and cortisol to dex suppression testing can be diagnostic of the source of androgen excess. Treatment, including oral contraceptives, low-dose glucocorticoids, and antiandrogens, should be chosen according to the patient's symptoms and source of androgens and should be combined with traditional therapy for the specific dermatologic disorder. PMID- 8243446 TI - Growth hormone in adolescence. Normal and abnormal. AB - The adolescent growth spurt is one of the dramatic physical changes that accompanies pubertal development. The rate of growth during the adolescent growth spurt is greater than at any other time of life after infancy. In this article, the pathophysiology of normal adolescent growth is reviewed, factors that control growth in adolescence are addressed, and the growth hormone deficiency syndromes presenting in adolescence are discussed. PMID- 8243447 TI - Diabetes mellitus during adolescence. AB - Poorly controlled diabetes may affect the tempo and course of pubertal growth and development. Pubertal changes induce glucose metabolism or specific insulin resistance. Because puberty greatly increases the risk of diabetes complications, the management offered at this transitional age is critically important. An understanding of the environmental and developmental influences on diabetes control and the effect of the physiologic changes of puberty improves the approach to diabetes management in the adolescent with diabetes. PMID- 8243448 TI - Disorders of calcium and phosphorus metabolism in adolescents. AB - During adolescence, there are marked changes in the metabolism of calcium and phosphorus and a dramatic increase in the rate of bone mineralization under the influence of the sex hormones, growth hormone, and insulin-like growth factor-1. More than 50% of adult bone mass is accumulated during puberty; failure to achieve maximum bone mineralization at this time may lead to osteopenia and its complications in later adulthood. This article discusses the causes, evaluation, and management of adolescents with hypocalcemia, hypercalcemia, and disorders of bone mineralization. PMID- 8243449 TI - Goiter in adolescents. AB - Enlargement of the thyroid gland during adolescence should be considered a pathologic rather than physiologic process. With diffuse enlargement in an asymptomatic patient, thyroid function tests and thyroid antibodies usually are the only tests needed to define the diagnosis of euthyroid autoimmune thyroiditis. Patients with lobular or nodular thyroid enlargement may require additional tests if the diagnosis of Hashimoto's thyroiditis cannot be established by the presence of thyroid antibodies in serum. The tests to define the anatomic and functional status of nodular thyroid disease include ultrasonography and radionuclide scintigraphy of the thyroid, but rarely fine needle biopsy. The only indications for surgical therapy of the thyroid are hyperfunctioning thyroid adenomas, the suspicion of thyroid carcinoma and Graves' disease in patients who are not responsive to antithyroid drug therapy and who are poor candidates for radioiodine ablative therapy. The prognosis of thyroid disease during c adolescence is usually excellent. PMID- 8243450 TI - Effects of irradiation on the thyroid gland. AB - External irradiation to the thyroid, even at low doses, induces subsequent development of thyroid nodules and carcinomas, with an estimated incidence of five per one million people per year if they have been exposed to 1 rad. Larger doses are associated with hypothyroidism. External irradiation also induces thyroid hyperplasia and is associated with an increased incidence of thyroid autoimmunity and the development of thyroiditis and Graves' disease. 131I, used in the treatment of Graves' disease and thyroid cancer for nearly 50 years, has not been associated with an increased incidence of thyroid cancer. Nevertheless, caution is advisable in use of radioiodide therapy in adolescents. Low levels of fallout from nuclear explosions probably have minimal, if any, effect on the thyroid; but recent high-level exposure in individuals near Chernobyl is reported to be associated with the development of thyroid malignancies in children. PMID- 8243451 TI - Effects of chemotherapy and irradiation on the gonads. AB - As the survival of children with cancer improves, attention must now be focused on understanding and preventing long-term toxicities of treatment and improving the quality of survival. Gonadal toxicity is relatively common, particularly with radiation therapy and alkylating agents. Continued research into the pathogenesis and prevention of gonadal toxicities needs to be included in future treatment strategies for childhood cancer. This will require a multidisciplinary approach with input from pediatric hematologist/oncologists, radiation oncologists, and pediatric endocrinologists. PMID- 8243452 TI - The therapy for hypothalamic-pituitary tumors. AB - Pituitary-hypothalamic tumors may profoundly affect endocrine functions. Although these are generally rare tumors of the central nervous system, they prominently figure into the differential diagnosis of children and adolescents with disorders of growth or puberty. PMID- 8243453 TI - Endocrine problems of adolescent pregnancy. AB - A number of changes in renal and endocrine physiology occur during pregnancy that alter hormone levels and affect a number of disease processes. Increased glomerular filtration causes an increase in hormone and substrate clearance. Increased placental steroid production causes an increase in hormone-binding globulin production, insulin resistance, and prolactinoma growth. Production of peptide hormones may cause changes in normal physiology and alter dynamic hormone testing. Placental vasopressinase increases vasopressin clearance. A number of diseases of hormone overproduction and underproduction affect pregnancy outcome and must be treated promptly by therapeutic modalities that also may affect the fetus. PMID- 8243454 TI - Nutrition in adolescence. AB - Adolescence is often considered a nutritionally vulnerable period because of its characteristic rapid physical and psychological changes. Nutritional risks for both undernutrition and overnutrition are present and may be increased during physiologic stress such as athletics and pregnancy. Appropriate nutrition is important for maintaining optimal health and normal growth and development, as well as for preventing future chronic disease. PMID- 8243455 TI - The endocrinology of stress and stress system disorders in adolescence. AB - The endocrinology of stress and the stress response is reviewed briefly. The interrelations of the stress system with various other endocrine axes and how this system may influence adolescent development and its various disorders are discussed. PMID- 8243456 TI - Protein glycosylation. Structural and functional aspects. AB - During the last decade, there have been enormous advances in our knowledge of glycoproteins and the stage has been set for the biotechnological production of many of them for therapeutic use. These advances are reviewed, with special emphasis on the structure and function of the glycoproteins (excluding the proteoglycans). Current methods for structural analysis of glycoproteins are surveyed, as are novel carbohydrate-peptide linking groups, and mono- and oligo saccharide constituents found in these macromolecules. The possible roles of the carbohydrate units in modulating the physicochemical and biological properties of the parent proteins are discussed, and evidence is presented on their roles as recognition determinants between molecules and cells, or cell and cells. Finally, examples are given of changes that occur in the carbohydrates of soluble and cell surface glycoproteins during differentiation, growth and malignancy, which further highlight the important role of these substances in health and disease. PMID- 8243457 TI - Refined crystal structure of phycoerythrin from Porphyridium cruentum at 0.23-nm resolution and localization of the gamma subunit. AB - The three-dimensional structure of the light-harvesting pigment-protein b phycoerythrin from the red alga Porphyridium cruentum has been determined at 0.23 nm resolution. The b-phycoerythrin structure is very similar to the structure of B-phycoerythrin from Porphyridium sordidum. Besides three non-identical residues there are only small differences between b-phycoerythrin and B-phycoerythrin alpha and beta subunits, respectively. In the crystals b-phycoerythrin forms an (alpha beta)6 hexamer (molecular mass: 236 kDa), whereas B-phycoerythrin additionally contains a 30-kDa gamma subunit. The comparison of the b phycoerythrin and B-phycoerythrin electron-density maps clearly reveals, that the gamma subunit is located inside the (alpha beta)6 aggregate. PMID- 8243458 TI - Characterization of the post-translational modifications in tubulin from the marginal band of avian erythrocytes. AB - Tubulin purified from turkey erythrocytes was characterized by partial protein sequence data, high-resolution IEF and by its reaction with antibodies specific for certain post-translational modifications. The tubulin from the marginal band contains a single alpha and beta isotype, i.e. alpha 1 and beta 6. Partial protein sequences and immunoblotting with antibody 6-11B-1 show that erythrocyte alpha 1 tubulin is not acetylated at Lys40. The acidic carboxy-terminal peptides purified by Mono Q chromatography and reverse-phase HPLC were characterized by sequence analysis and mass spectrometry. Although erythrocyte alpha tubulin is almost completely detyrosinated it retains the penultimate glutamic acid residue, which is partially lost in brain tubulin. Thus erythrocyte tubulin is an excellent substrate for extensive in vitro tyrosination by tubulin-tyrosine ligase. Erythrocyte alpha and beta tubulin lack the side-chain polyglutamylation found in all major tubulins from adult brain. Finally we show that about 10% of the beta tubulin is phosphorylated at Ser441. Thus erythrocyte tubulin is an unusual homogeneous preparation. It contains the minimum possible number of tubulin isotypes and the only post-translational modifications detected (detyrosination and phosphorylation) are reversible. PMID- 8243459 TI - Mouse gelatinase B. cDNA cloning, regulation of expression and glycosylation in WEHI-3 macrophages and gene organisation. AB - Gelatinase B is a regulated matrix metalloproteinase with an important role in the remodelling of extracellular matrices and of basement membranes. To study the structure and function of gelatinase B in the mouse, the cDNA was cloned from a macrophage cell line (WEHI-3). Using this cDNA, a cosmid clone with the mouse gene was isolated. The complete gene (8 kbp) was sequenced and compared with the human gene structure. There was 78% similarity at the cDNA level and the exon/intron structure of the murine gene was similar to the human counterpart. At the 5' untranslated side, 1200 bp of the promoter/enhancer region were sequenced and found to contain several transacting-factor-binding sites. The mRNA transcription-initiation site was determined by non-isotopic primer-extension analysis. Polymerase-chain-reaction amplification of cDNAs yielded indirect evidence for a reverse-transcription stop in WEHI-3 cell mRNA. The DNA-derived mouse-protein structure exhibited 82% similarity with the human one. This similarity was functionally reflected by cross-reactivity of the mouse protein with an antiserum against human gelatinase B. The production of murine gelatinase B was studied at the protein level by zymography and at the mRNA level by Northern blot analysis. In WEHI-3 cells the gelatinase B protein is induced by bacterial lipopolysaccharide, phorbol ester, double-stranded RNA and the cytokine interleukin-1. Regulation of activity and structural heterogeneity of gelatinase B in WEHI-3 cells were shown to occur at the gene regulatory level, by expression of the matrix metalloproteinase inhibitor TIMP-1, and by glycosylation of the secreted protein. PMID- 8243460 TI - Determinants of the brain-specific expression of the rat aldolase C gene: ex vivo and in vivo analysis. AB - A 115-bp promoter fragment of the aldolase C gene is sufficient for conferring neural cell specificity on a reporter gene, in cultured PC12 cells and in transgenic mice. In vitro DNase I protection experiments detected two footprints on the promoter, termed boxes A/A', and B. The 5' A/A' box contains overlapping Sp1 and Krox20/Krox24 binding sites; it binds Sp1 in fibroblasts (box A') and a different complex in brain (box A). Any deletion or mutation of this box that impairs protein recognition also suppresses promoter activity. The replacement of box A/A' by a Sp1 consensus binding site results in the loss of the brain specificity of expression in transgenic mice. Further 3', box B is composed of a 5' direct repeat and a 3' GC box consisting of overlapping Sp1 and Krox20/Krox24 binding sites. Mutation of the direct repeat subregion appears to be more deleterious for the promoter activity than mutation of the G+C-rich subregion. PMID- 8243461 TI - Identification of O-linked oligosaccharide chains in the activation peptides of blood coagulation factor X. The role of the carbohydrate moieties in the activation of factor X. AB - Conversion of factor X to factor Xa results in release of a heavily glycosylated activation peptide. Analysis of protease-digested glycopeptides derived from the activation peptides of bovine and human blood coagulation factor X allowed the identification of sites of the O-linked oligosaccharide chains in these peptides. Glycopeptides were prepared from the activation peptides by digestion with chymotrypsin or Staphylococcus aureus V8 protease. By combined analysis of amino acid sequence and sialic acid content, we found that bovine factor X had an O linked oligosaccharide chain linked to Thr26, and human factor X had four carbohydrate-attachment sites, namely, O-glycosidic linkages to Thr17 and Thr29, respectively, and N-glycosidic linkages to Asn39 and Asn49, respectively, in their activation peptides. The O-linked carbohydrate-attachment sites were identified since the yields of phenylthiohydantoin derivatives of amino acids that corresponded to their residues were increased during amino acid sequencing after deglycosylation of the glycopeptides with sialidase and O-glycanase. The effect of deglycosylation of bovine factor X1 was investigated with factor-X activating enzyme from Russell's viper venom or extrinsic Xase (factor VIIa/tissue factor/phospholipid) by examining the activation rates of derivatives of factor X prepared using O-glycanase, sialidase, and/or N-glycanase. The removal of O-linked carbohydrate resulted in a decrease in the rate of activation. It appears that carbohydrate residues in factor X play an important role in the activation of the zymogen. PMID- 8243462 TI - Investigation of the oxygenation of phospholipids by the porcine leukocyte and human platelet arachidonate 12-lipoxygenases. AB - When arachidonate 12-lipoxygenase purified from porcine leukocytes was incubated aerobically with 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine, the phospholipid reacted at up to 30% of the rate of a free fatty acid substrate; the esterified arachidonic acid was oxygenated predominantly to the (12S)-12 hydroperoxy product. The porcine leukocyte enzyme was also capable of metabolizing phosphatidylcholine containing esterified (15S)-15-hydroperoxy 5,8,11,13-eicosatetraenoic acid; oxygenation occurred predominantly at the 14R position. Reaction with mitochondrial and endoplasmic membranes of rat liver produced esterified (12S)-12-hydroperoxy-5,8,10,14-eicosatetraenoic acid and (13S)-13-hydroperoxy-9,11-octadecadienoic acid as major oxygenation products. Thus, porcine leukocyte 12-lipoxygenase is capable of oxygenating not only free polyenoic fatty acids but also more complex substrates such as phospholipids and biomembranes. In contrast, the human platelet 12-lipoxygenase is almost inactive with these esterified polyenoic fatty acids. In regard to the function of these enzymes, the leukocyte-type of 12-lipoxygenase has similar catalytic activities to the mammalian 15-lipoxygenase and its physiological function may include the structural modification of membrane lipids. PMID- 8243463 TI - Complete amino acid sequences of five dimeric and four monomeric forms of metallothionein from the edible mussel Mytilus edulis. AB - Cadmium-induced metallothioneins from the common sea mussel, Mytilus edulis, were shown to comprise of two groups of isoforms having apparent molecular masses of 10 kDa and 20 kDa. The 10-kDa group was resolved by anion-exchange chromatography into four fractions while the 20-kDa group was resolved into three fractions using this method. After metal removal and S-methylation of the cysteine residues using methyl-p-nitrobenzenesulphonate the complete amino acid sequences were determined. Five isoforms of the 20-kDa group were shown to possess monomeric units consisting of 71 amino acids. These proteins were distinct from the four 72 amino-acid proteins of the 10-kDa group. The FASTA algorithm has been used to compare the degree of similarity between the mussel metallothionein MT-10-IV isoform and other metallothioneins. The mussel MT-10-IV isoform exhibited substantial similarity to other molluscan metallothioneins. Moreover, the mussel metallothionein exhibited more similarity to vertebrate metallothioneins than to those of non-molluscan invertebrates, thus suggesting that the mussel metallothioneins are class I metallothioneins. PMID- 8243464 TI - Anomalous interaction of Sp1 and specific binding of an E-box-binding protein with the regulatory elements of the Na,K-ATPase alpha 2 subunit gene promoter. AB - We identified cis elements in the 5'-flanking region of rat Na,K-ATPase alpha 2 subunit gene (Atp1a2) using transient transfection assays in L6 rat skeletal muscle myoblast cells. By 5'-deletion mutation analysis, the region between nucleotide positions -175 and -108 was identified as a positive regulatory region. In the region, the distal E box (nucleotides -144 to -139) acts as a negative regulatory element, and the Sp1 consensus sequence (nucleotides -123 to 118) and the GGGAGG sequence (nucleotides -114 to -109) act as positive regulatory elements. Gel-retardation analysis revealed that binding factors are an E-box-binding protein and Sp1. DNase I footprinting and methylation interference analyses revealed that Sp1 binds to the region from nucleotides -122 to -101 and the E-box-binding protein to the region from nucleotides -144 to 136. T4 DNA polymerase footprinting revealed that there are three Sp1-binding sites in the region and that Sp1 binds to one of the three sites in a mutually exclusive manner. The mechanism by which Sp1 activates the Atp1a2 promoter is discussed. PMID- 8243465 TI - Recombinant coho salmon insulin-like growth factor I. Expression in Escherichia coli, purification and characterization. AB - Recombinant coho salmon insulin-like growth factor I (rsIGF-I) was produced in Escherichia coli, purified and characterized. The rsIGF-I expression vector was constructed by polymerase chain reaction and cloning into a plasmid containing a phage T7 RNA polymerase promoter. The rsIGF-I was recovered from bacterial inclusion bodies, solubilized under reducing conditions, immediately refolded, then fractionated by a two-step ion-exchange chromatography on DEAE-52 and Mono-S columns. It was further purified by HPLC on a reverse-phase Asahi-Pak C4P-50 C4 column. Purification of rsIGF-I was monitored by SDS/PAGE and immunoblot with anti-[human somatomedin C (SM C)/IGF-I] serum. The rsIGF-I appeared as a single band with molecular mass of 7 kDa, the same size as recombinant human IGF-I (rhIGF-I) and cross-reacted with anti-(human SM C/IGF-I) serum. The amino acid sequence of rsIGF-I contained an NH2-terminal methionine residue followed by the sequence predicted for mature sIGF-I. At concentrations in the range 3.9-250 ng/ml, rsIGF-I significantly stimulated sulfate uptake by the cultured branchial cartilage of coho salmon. The stimulatory effect of rsIGF-I was concentration dependent and slightly more potent than that of rhIGF-I at the highest concentration tested. PMID- 8243466 TI - Structure and dynamics of the acidic compact state of apomyoglobin by frequency domain fluorometry. AB - The conformational dynamic properties of tuna apomyoglobin, a single tryptophan containing protein, in the acidic compact state, as well as in the native and in the fully unfolded state, have been explored by frequency-domain fluorometry. Apomyoglobin at acidic pH in the presence of high salt concentration displays bimodal tryptophanyl lifetime distributions which may be related to the simultaneous presence of different populations of structural states (compact and fully unfolded states). The tryptophanyl anisotropy decay indicated that the acidic compact state displays at least two rotational correlational times, suggesting that this state possesses a complex geometrical organization. 1 Anilino-8-naphthalene sulfonate (ANS), bound both to native and compact protein forms, shows broad unimodal lifetime distributions. The small time dependence of the ANS emission spectra indicated that the solvent dipolar reorganization are either absent or they occur on a time scale much shorter than the lifetime of the excited ANS molecule bound to apomyoglobin. The anisotropy decay data relative to the extrinsic fluorophore (ANS) are consistent with the presence of a single rotational correlation time for both native (12.1 ns) and compact (6.2 ns) states. PMID- 8243467 TI - Non-cooperative effects of glucose and 2-deoxyglucose on their metabolism in Saccharomyces cerevisiae studied by 1H-NMR and 13C-NMR spectroscopy. AB - The reciprocal effects of 2-deoxy-D-glucose (dGlc) and glucose (Glc) on the aerobic metabolism of Glc and dGlc in Glc-grown repressed Saccharomyces cerevisiae were studied at 30 degrees C in a standard pyrophosphate medium containing about 4.5 x 10(7) cells/ml. 1H-, 13C-NMR and biochemical techniques were used for quantitative evaluation of Glc and dGlc metabolites. The detection of intracellular dGlc and the determination of the intracellular dGlc6P/dGlc ratio were realised using [1-13C]dGlc and 13C-NMR spectroscopy. The rates of Glc consumption in the absence and in the presence of 5 mM dGlc and 20 mM dGlc were 29 +/- 0.03 mM/min (n = 3), 16 +/- 0.02 mM/min (n = 3), and 0.08 +/- 0.01 mM/min (n = 3), respectively. This means that the Glc consumption is reduced about 41% and 70% in the presence of 5 mM and 20 mM dGlc, respectively. When dGlc is the unique carbon source, only alpha and beta anomers of dGlc6P were formed. Their quantities are equivalent and reach the maximum values within 1 h of incubation and then decrease gradually. By contrast, Glc favours the consumption of dGlc and the synthesis of dGlc6P, dideoxy-trehalose (dGlc-dGlc), deoxy-trehalose (dGlc Glc). In the presence of Glc, dGlc6P reaches a plateau after 1 h or 2 h of incubation while the quantities of trehalose (Glc-Glc), dGlc-dGlc, dGlc-Glc, which are small at 1 h, rapidly increase with time. The reasons why dGlc and Glc exert opposite effects on their metabolism are discussed. The production of Glc Glc decreases with increasing the external dGlc concentration or the dGlc/Glc ratio. The effect of dGlc on the biosynthesis of Glc-Glc can be explained by the competition of dGlc and Glc with respect to hexokinase. Although Glc favours the synthesis of dGlc6P, the maximum concentration of dGlc6P shows little dependence on the external dGlc concentration as long as glucose is not exhausted. The internal dGlc6P/dGlc ratio at equilibrium, about 4.7 +/- 0.7, is also found to be independent of the dGlc concentration in the suspension. Only a small fraction of dGlc6P disappears to give rise to the formation of dGlc-dGlc and dGlc-Glc. At equilibrium the inverse reaction from dGlc6P to dGlc may be important to compensate for the fast reaction of dGlc phosphorylation by hexokinase. At least nine series of experiments were conducted and showed that, in pyrophosphate media and for incubation times less than 4 h, dGlc-dGlc was not observed in the absence of Glc.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243468 TI - Effect of lysine ionization on the structure and electrochemical behaviour of the Met44-->Lys mutant of the blue-copper protein azurin from Pseudomonas aeruginosa. AB - The structural and spectrochemical effects of the replacement of Met44 in the hydrophobic surface patch of azurin from Pseudomonas aeruginosa by a lysine residue were studied as a function of the ionization state of the lysine. In the pH range 5-8, the optical absorption, resonance Raman, EPR and electron spin-echo envelope modulation spectroscopic properties of wild-type and Met44-->Lys (M44K) azurin are very similar, indicating that the Cu-site geometry has been maintained. At higher pH, the deprotonation of Lys44 in M44K azurin (pKa 9-10) is accompanied by changes in the optical-absorption maxima (614 nm and 450 nm instead of 625 nm and 470 nm) and in the EPR gII value (2.298 instead of 2.241), indicative of a change in the bonding interactions of Cu at high pH. The strong pH dependence of the electron self-exchange rate of M44K azurin supports the assignment of Lys44 as the ionizable group and demonstrates the importance of the hydrophobic patch for electron transfer. The pH dependence of the midpoint potentials of wild-type and M44K azurin can be accounted for by the ionizations of His35 and His83 and by the additional electrostatic effect of the mutation. PMID- 8243469 TI - Effects of glycosylation on protein structure and dynamics in ribonuclease B and some of its individual glycoforms. AB - In an attempt to elucidate the role of carbohydrates on protein structure and dynamics in glycoproteins, ribonuclease B (RNase B), containing a single glycosylation site at Asn34, has been investigated and compared with the enzyme in the unglycosylated form (RNase A). RNase B consists of five glycoforms: Man5GlcNAc2, Man6GlcNAc2, Man7GlcNAc2, Man8GlcNAc2 and Man9GlcNAc2 (Man5 9GlcNAc2). The individual glycoforms Man1GlcNAc2 (synthetic) and Man5GlcNAc2 of RNase B have been studied to ascertain any specific effects of the different oligosaccharides. NMR measurement of amide-proton/deuterium exchange rates shows that glycosylation of the enzyme leads to the protection of amide-proton resonances from solvent exchange for a large number of residues, both in the vicinity of the glycosylation site (e.g. residues 29-34 and 35) and remote from it (e.g. residues 57-61 and 75-76). In addition, residues 10-13, 32, 34 and 35, which are observed to be protected from exchange as a result of glycosylation in the individual glycoforms Man1Glc-NAc2-RNase and Man5GlcNAc2-RNase (when compared with RNase A) are less protected in RNase B. This additional protection in the glycoforms Man1GlcNAc2-RNase and Man5GlcNAc2-RNase may arise from steric hindrance between the oligosaccharide and protein reducing solvent accessibility. The rates of solvent exchange of amide protons for residues 10-13, 32, 34 and 35 are dependent on the oligosaccharide moiety. The average amide-proton/deuterium exchange rate in Man6-9Glc-NAc2-ribonucleases for residues 10-13 and 35 is approximately three times greater than Man5Glc-NAc2-ribonuclease, while for residues 32 and 34 it is approximately 7-11 times greater. CD analysis of RNase A and RNase B revealed the carbohydrate moiety to have a small stabilizing effect (approximately 5 kJ/mol) on the protein. PMID- 8243470 TI - Gly387 of murine ornithine decarboxylase is essential for the formation of stable homodimers. AB - In its active form mammalian ornithine decarboxylase (ODC) is a homodimer composed of two 53-kDa subunits while the monomer retains no enzymic activity. In the present study we demonstrate that Gly387 of mouse ODC plays an important role in enabling dimer formation. Gly387 of mouse ODC, an evolutionary conserved residue, was converted to all possible 19 amino acids using site-directed mutagenesis. With the exception of alanine, all other substitutions of Gly387 completely abolished enzymic activity. Cross-linking analysis and fractionation through a Superose-12 sizing column have demonstrated that mutant subunits are detected only in their monomeric form. These results strongly suggest that the primary lesion of substitution at position 387 of mouse ODC is the inability of mutant subunits to associate with each other to form the active homodimers. In agreement with this conclusion, G387A, the only mutant that retained partial activity, displayed reduced dimerization. The degradation rate of ODC mutants in which Gly387 was substituted by aspartic acid or alanine was enhanced compared to the wild-type enzyme, suggesting that monomers may be more susceptible to degradation. PMID- 8243471 TI - Structure/activity relationships in porphobilinogen oxygenase and horseradish peroxidase. An analysis using synthetic hemins. AB - The apo-enzymes of porphobilinogen oxygenase and horseradish peroxidase were reconstituted with hemin IX, deuterohemin IX, 2,4-diacetyldeuterohemin IX, 2 vinyl-4-deuterohemin IX and hemin I. The apoproteins did not reconstitute with the dimethyl or diethyl esters of hemin IX. The native enzymes and the synthetic hemoproteins showed similar oxygenase activities toward porphobilinogen in the presence of dithionite and oxygen. They also showed peroxidase activity in the presence of H2O2, which was affected by the side-chain substitution pattern of the hemes. Oxygenase activities, however, were not affected by the heme structure. Iron chelators completely inhibited the oxygenase, but not the peroxidase activities. The EPR spectra of the native and synthetic porphobilinogen oxygenase showed that dithionite reduction produced a rapid disappearance of the high-spin heme-iron signal at g = 6.0. It reappeared 1 min later but the enzyme retained its catalytic activity. The changes in the EPR spectra could be correlated with the biphasic kinetics of the oxygenase reaction which was very fast during the first minute and then decreased to a half-value rate. The oxygenase reaction was inhibited by addition of superoxide dismutase during the fast rate phase, but not during the slower phase. These results could be explained by the formation of a superoxide anion during the first minute of the oxygenase reaction, after which a protein-stabilized radical (g = 2.0) is generated (very likely a tyrosyl radical). The latter then oxidizes the substrate porphobilinogen and facilitates its reaction with O2 to give oxopyrrolenines. PMID- 8243472 TI - Reconstitution of holotransketolase is by a thiamin-diphosphate-magnesium complex. AB - When human erythrocyte apo-transketolase is mixed with cofactors and substrates, the progress curve exhibits a lag phase. Elimination of the lag phase requires the presence of saturating concentrations of cofactors, thiamin diphosphate and Mg2+. The most simple explanation of the observed hysteretic transition is that the slow binding of a Mg(2+)-thiamin-diphosphate species precedes slow isomerisation of the enzyme to the active form. Although the hysteretic transition involves more than one process, it does not involve the dissociation association of enzyme subunits. The best estimate of the apparent Km, 1.59 +/- 0.23 microM, for the binding of Mg(2+)-thiamin diphosphate to transketolase was obtained in the presence of a high non-inhibitory concentration of magnesium and varied concentrations of thiamin diphosphate. Thus the reconstitution of the human enzyme differs from the yeast enzyme, which undergoes a rate-limiting dimerisation during reconstitution. PMID- 8243473 TI - The separate roles of plant cis and trans prenyl transferases in cis-1,4 polyisoprene biosynthesis. AB - In plants, the elongation of cis-1,4-polyisoprene (natural rubber, M(r) > 10(6) requires a small transallylic diphosphate (< or = C20) initiator. The trans allylic diphosphates are hydrophilic cytosolic compounds, whereas cis-1,4 polyisoprene is hydrophobic and compartmentalised in subcellular rubber particles. In this paper, it is demonstrated that soluble trans-prenyl transferase from latex of Hevea brasiliensis functions solely as farnesyl diphosphate synthase, and plays no direct role in cis-1,4-polyisoprene elongation. The cis-1,4-prenyl transferase is firmly associated with the H. brasiliensis rubber particle, as is also the case in other rubber-producing species [Archer, B. L., Audley, B. G., Cockbain, E. G. & McSweeney, G. P. (1963) Biochem. J. 89, 565-574; Madhavan, S., Greenblatt, G. A., Foster, M. A. & Benedict, C. R. (1989) Plant Physiol. 89, 506-511; Siler, D. J. & Cornish, K. (1993) Phytochemistry 32, 1097-1102]. The experimental data explain and refute previous reports in which soluble trans-prenyl transferase isolated from H. brasilensis latex was attributed both trans-prenyl transferase and cis-prenyl transferase activities [Light, D. R. & Dennis, M. S. (1989) J. Biol. Chem. 264, 18589-18597; Light, D. R., Lazarus, R. A. & Dennis, M. S. (1989) J. Biol. Chem. 264, 18598-18607]. Thus, it appears that plant prenyl transferases are comparable to animal enzyme systems in which trans-prenyl transferases are soluble enzymes whilst cis-prenyl transferases are membrane-bound [Ericcson, J., Runquist, M., Thelin, A., Andersson, M., Chojnacki, T. & Dallner, G. (1992) J. Biol Chem. 268, 832-838]. PMID- 8243474 TI - Phenotype of recombinant Leishmania donovani and Trypanosoma cruzi which over express trypanothione reductase. Sensitivity towards agents that are thought to induce oxidative stress. AB - Trypanothione reductase is thought to be important in maintaining an intracellular reducing environment in trypanosomatids. To investigate the role of trypanothione reductase we transfected Leishmania donovani and Trypanosoma cruzi with an expression vector containing the L. donovani trypanothione reductase gene and achieved over-expression of enzyme activity (10-14-fold) in transformed cells. Following treatment of L. donovani cells with the thiol-oxidizing agent diamide, the ability to regenerate dihydrotrypanothione from trypanothione disulphide was considerably enhanced in cells which over-expressed trypanothione reductase. However, the growth of transformed and control cells was equally sensitive to inhibition by nifurtimox, nitrofurazone and gentian violet, drugs that are thought to act by inducing oxidative damage. Likewise, growth of transformed and control cells were equally susceptible to inhibition by hydrogen peroxide, and control and transformed L. donovani promastigotes metabolized hydrogen peroxide at comparable rates. Thus, these experiments suggest that the ability to regenerate dihydrotrypanothione from trypanothione disulphide is not a rate-limiting step in the metabolism of hydrogen peroxide. PMID- 8243475 TI - Increased phosphorylation of eukaryotic initiation factor 4 alpha during early activation of T lymphocytes correlates with increased initiation factor 4F complex formation. AB - Mature porcine peripheral blood mononuclear cells (PPBMCs) exist in a resting state both in vivo and when maintained in culture, with low translation rates consistent with their non-proliferative state. When cultured in the presence of the appropriate mitogen, there is a 2-4-fold increase in the rate of protein synthesis per ribosome within 4 h of stimulation [Kay, J. E., Ahern, T. and Atkins, M. (1971) Biochim. Biophys. Acta 247, 322-334]. Studies on extracts prepared from unstimulated cells have suggested lesions in initiation factor activity, primarily affecting the binding of mRNA to ribosomes [Ahern, T., Sampson, J. and Kay, J. E. (1974) Nature 248, 519-521]. In these studies, we have demonstrated that activation of quiescent PPBMCs with the phorbol ester phorbol 12-myristate 13-acetate or concanavalin A leads to a rapid 2-4-fold increase in the rate of protein synthesis within 1 h or 4 h, respectively, which is insensitive to the transcriptional inhibitor, 5,6-dichlorobenzimidazole riboside. Relative to control cells, both phorbol ester and concanavalin A induce a 2-4 fold increase in labelling of the eukaryotic initiation factor eIF-4 alpha with phosphate in vivo, which primarily reflects a small net increase in phosphorylation rather than phosphate turnover on eIF-4 alpha. Similarly, with the human leukaemic T cell line JURKAT, stimulation of the T cell receptor with the monoclonal antibody, OKT-3, or treatment with phorbol ester induces a 2-3 fold increase in eIF-4 alpha phosphorylation within 30 min. Analysis of phosphorylation by two-dimensional gel electrophoresis and measurement of kinase activity towards synthetic peptides, indicate that this increased labelling also reflects increased eIF-4 alpha kinase activity rather than phosphate turnover on eIF-4 alpha. Of central importance is the finding that, concomitant with increased rates of protein synthesis following stimulation of PPBMCs with either phorbol ester or concanavalin A, there is a significant increase in the level of eIF-4 alpha recovered in high-molecular-mass complexes. These data suggest that, in quiescent PPBMCs, eIF-4F may be limiting and that the association of eIF-4 alpha and eIF-4 gamma into high-molecular-mass complexes is regulated by phosphorylation and may play a pivotal role in translational control. PMID- 8243476 TI - Sequence and expression of the gene encoding the respiratory nitrous-oxide reductase from Paracoccus denitrificans. New and conserved structural and regulatory motifs. AB - The structural gene for the respiratory nitrous-oxide reductase from Paracoccus denitrificans has been cloned using a probe derived from the structural gene, nosZ, for this enzyme from Pseudomonas stutzeri. The cloned gene could be expressed surprisingly well (presumably yielding an apo-protein) using an expression vector in Escherichia coli. Sequencing the nosZ gene from P. denitrificans has shown that the periplasmic nitrous-oxide reductase of this organism is highly similar in sequence to previously derived primary sequences for the enzyme from three other organisms. As with the other reductases, an unusually long signal sequence is deduced and a common motif of GXXRRXXLG near the beginning of this sequence is present. The results of N-terminal sequencing of the mature nitrous-oxide reductase from the closely related organism Thiosphaera pantotropha indicate that processing of the P. denitrificans precursor occurs between amino acids at positions 57 and 58. The predicted signal peptide is therefore of the same length and of similar overall structure to that previously described for the P. denitrificans methylamine dehydrogenase small subunit (MauA). The P. denitrificans sequence for the mature nitrous-oxide reductase reduces from 14 to 11 and 6 to 4, respectively, the number of conserved histidine and methionine residues compared to previous sequences. Three cysteine and four tryptophan residues, previously identified as conserved amongst nitrous oxide reductases, are found in the Paracoccus enzyme. A comparison of the sequence of the C-terminal region of the nitrous-oxide-reductase sequence with that for the CuA region of subunit II of the cytochrome aa3 from P. denitrificans reveals considerable sequence similarities. Upstream of the structural gene for nosZ are sequences TTGAAGCTTAACCAG (centred at position -21 with respect to the start codon) and CCCGGTGGTCATCAAG (centred at position -126). Although both could be FNR (ANR) boxes, the latter is far more probable to have this role because only it is likely to be upstream of a promoter site. This is the first indication at the DNA sequence level for the existence of this regulatory system in P. denitrificans. Analysis of the flanking DNA sequences revealed reading frames upstream and downstream of the nosZ gene showing similarity to the nosR and nosD genes, respectively, of Pseudomonas species. An S30 in vitro transcription/translation system was developed for P. denitrificans which permitted the expression of the cloned gene for nitrous-oxide reductase and which will be of general value in other studies of this organism. PMID- 8243477 TI - Structure/activity relationship of leukotriene B4 and its structural analogues in chemotactic, lysosomal-enzyme release and receptor-binding assays. AB - The biological activities of chemically synthesized leukotriene B4 and eight structural analogues have been studied using chemotaxis, lysosomal-enzyme release and receptor-binding assays on human neutrophils. The results show that increasing the number of double bonds between C14 and C20, having triple bonds at C6 or C14, substitution of the primary carboxyl group at C1, changing the geometry of the double bond at C6 from the cis to trans configuration and changing the chirality of the hydroxyl group at C12 from the R to the S configuration result in substantial loss of both biological activity and the capacity to bind to the LTB4 recognition site in parallel. We suggest that the functional epitopes of 5S,12R-dihydroxy-6,14-cis-8,10-trans-icosatetraenoic acid (LTB4) are either the same, or reside in the same domain as the binding site for the LTB4 receptor. Development of LTB4 antagonists to the high-affinity LTB4 receptor, based on the structure of LTB4, is unlikely to be successful. PMID- 8243478 TI - Partial purification and characterization of a circulating hypertensive factor in spontaneously hypertensive rats. AB - Parabiosis and cross-circulation experiments with spontaneously hypertensive and normotensive rats gave indications for a previously unidentified circulating hypertensive agent. In this study, plasma from normotensive and hypertensive rats was fractionated and the vasopressor action of the corresponding fractions was measured in the isolated perfused rat kidney. One of three vasoactive fractions obtained by gel filtration (Biol-Gel P2) from hypertensive rats showed a significantly higher activity (increase in perfusion pressure by 1502.9 +/- 438.9 Pa) than that from normotensive rats (increase in perfusion pressure by 505.4 +/- 186.2 Pa, P < 0.01). Further chromatographic separations of this fraction revealed that the hypertensive factor is hydrophilic and has no ionic groups or vicinal diol groups. The molecular mass was estimated by dialysis and the matrix assisted laser desorption/ionization mass spectrometry to be in the range of 1 kDa. The vasopressor is heat resistant and not degradable with trypsin or carboxypeptidase Y. The vasopressor action was not inhibited with the angiotensin II-receptor antagonist saralasin, the alpha-receptor antagonist phentolamine, the thromboxane-receptor antagonist carbocyclic thromboxane A2 or the serotonin antagonist ketanserin. The results confirm the existence of a vasopressor factor in the plasma of hypertensive rats and, in a lower concentration, of normotensive rats, which is possibly related to the pathogenesis of essential hypertension. The chromatographic behavior suggests that this factor is different from the parathyroid hypertensive factor described recently. PMID- 8243479 TI - Aminoacyl chloromethanes as tools to study the requirements of NADPH oxidase activation in human neutrophils. AB - Previous studies from this laboratory described the kinetic characteristics of the inhibition by tosylphenylalanine chloromethane (TosPheCH2Cl) on superoxide anion production by human neutrophils (PMN) stimulated with a phorbol ester (PMA). In this study we present further evidence concerning the potential role of the chloromethane target in the normal cellular activation of NADPH oxidase. When PMN are treated with TosPheCH2Cl and subsequently PMA, or with the two reagents in the reverse order, the inhibition of superoxide production by the intact cells is still present in a particulate NADPH oxidase fraction prepared from these cells. Nevertheless, when cells incubated only with the chloromethane and not with PMA are disrupted, both their cytosolic and membrane fractions are fully competent in the cell-free activation assay. Thus, the chloromethane target has a role in NADPH oxidase activation exclusively at the cellular level. This observation constitutes additional evidence in favour of the idea that activation in the cell-free system reflects only partially the events which occur in the cells. When cells are activated with PMA, their cytosol displays a loss of activating capacity in the cell-free activation assay in the presence of arachidonate, as was shown before with SDS as activator [Ambruso, D. R., Bolsher, B. G. J. M., Stockman, P. M., Verhoeven, A. J. & Roos, D. (1990) J. Biol. Chem. 265, 924-930]. This phenomenon was shown to arise most probably from the translocation of cytosolic factors to the membrane, resulting in a depleted cytosol. When superoxide production was inhibited by cell treatment with TosPheCH2Cl, either before or after activation with PMA, the cytosol from inhibited cells showed a recovery of activation capacity in the cell-free system. This effect probably results from TosPheCH2Cl inhibiting the translocation of the cytosolic factors when added before PMA. This results in an insufficient activation at the membrane level, which was previously considered as an inhibition. The effect of TosPheCH2Cl, when added after PMA, can best be explained again as an inhibition of translocation in the frame of the continuous replenishment-deactivation hypothesis proposed by Akard et al. [Akard, L. P., English, D. & Gabig, T. G. (1988) Blood 72, 322-327]. Thus, TosPheCH2Cl is apparently a promising new tool for studying the activation of NADPH oxidase at the cellular level.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243480 TI - Analysis of the DNA topoisomerase-II-mediated cleavage of the long terminal repeat of Drosophila 1731 retrotransposon. AB - The interaction of DNA topoisomerase II with the long terminal repeat (LTR) of the Drosophila melanogaster 1731 retrotransposon was studied. The covalent binding of topoisomerase II to the LTR was strongly stimulated by different inhibitors of the enzyme 4'-demethylepipodophyllotoxin-9-(4,6-O-2-ethylidene-beta D-glucopy ranoside (VP-16), 4'-(9-acridinylamino)methanesulfon-m-anisidine) (m AMSA) and an ellipticine derivative. Enzyme-mediated DNA cleavage could be observed in the absence of inhibitors and was stimulated in their presence. Cleavage occurred predominantly at sites located within or at the boundary of alternating purine/pyrimidine tracts in agreement with previous observations [Spitzner, J. R., Chung, I. K. & Muller, M. T. (1990) Eukaryotic topoisomerase II preferentially cleaves alternating purine-pyrimidine repeats, Nucleic Acids Res. 18, 1-11]. In addition, all of the cleavage sites observed in the absence of inhibitor were located in the U3 region of the LTR. The site specificity of drug induced cleavage was studied and the conformity of the cleavage sites with previously established consensus sequences was examined. Our results suggest that DNA topoisomerase II, through its ability to alter the degree of DNA supercoiling, might be involved in the control of different functions of the LTR. PMID- 8243481 TI - Failure to detect silent HIV infection by polymerase chain reaction in subjects at risk for heterosexually transmitted HIV type 1 infection. AB - The prevalence of silent HIV-1 infections in subjects at risk of acquiring HIV infection by heterosexual transmission was assessed using a nested polymerase chain reaction (PCR) assay. The two groups of risk subjects consisted of 92 female prostitutes and 43 heterosexual partners of infected individuals. Appropriate positive and negative control persons were included in the study. Serum samples were also tested for antibody to HIV-1 by an enzyme immunoassay (EIA) and positive results confirmed by Western blot. PCR results in the two risk groups and the positive and negative controls were in full agreement with serological results. It is concluded that silent infection with HIV-1 is infrequent in persons at risk for heterosexual transmission. PMID- 8243482 TI - Impact of screening donor blood for alanine aminotransferase and antibody to hepatitis B core antigen on the risk of hepatitis C virus transmission. AB - The transfusion-related risk of transmission of hepatitis C virus (HCV) was evaluated in France for the periods before and after exclusion of donor blood units with the surrogate markers elevated alanine aminotransferase (ALT) levels and antibody to hepatitis B core antigen (anti-HBc). A total of 1,412 blood recipients undergoing surgery were followed up prospectively in the period from 1986 to 1989. The stored serum samples were tested for antibodies to HCV by an enzyme immunoassay (EIA) and the result in reactive sera confirmed by a recombinant immunoblot assay (RIBA). The risk of HCV transmission was estimated by the maximum likelihood method for a subpopulation of 892 recipients divided into three groups. Of 55 (3.9%) EIA positive patients, 56.4% were found to be positive prior to transfusion. HCV seroconversion (positive RIBA) occurred in 22 patients (1.6%). The risk of HCV transmission per 1,000 transfused blood units decreased significantly from 4.11 in Group 1 (receiving non-screened blood) to 3.43 in Group II (receiving ALT screened blood) and to 1.40 in Group III (receiving ALT and anti-HBc screened blood). These results demonstrate that screening of donors for surrogate markers had reduced the risk of HCV transmission before the introduction of a systematic anti-HCV screening policy in France in March 1990. PMID- 8243483 TI - Survey of clinical isolates of Staphylococcus aureus for borderline susceptibility to antistaphylococcal penicillins. AB - On the basis of the MICs of methicillin and oxacillin, 975 clinical isolates of Staphylococcus aureus were categorized as having resistance, borderline susceptibility or full susceptibility to penicillinase-resistant penicillins (PRPs). The borderline phenotype accounted for 122 isolates (12.5%), whereas 562 isolates were fully susceptible and 290 resistant; one remaining isolate had resistance to methicillin and borderline susceptibility to oxacillin. Reductions in the MICs of methicillin and oxacillin in the presence of sulbactam were greater in strains with borderline PRP susceptibility than in fully susceptible or resistant isolates. Over 99% of fully PRP-susceptible strains, 93% with borderline susceptibility and 71% of resistant strains were susceptible to ampicillin/sulbactam. The production of beta-lactamase, assayed in all strains using nitrocefin as substrate, could be detected without prior induction in 729 strains and after induction only in another 156 strains. With only two exceptions, the beta-lactamase negative strains were part of the fully PRP susceptible group of organisms (88 of 562 isolates). Among the borderline isolates, strong beta-lactamase reactions were encountered with particular frequency, but not in all strains and not exclusively in borderline strains. Although associated with the majority of borderline strains, beta-lactamase hyperproduction thus did not appear to be an essential feature of the borderline phenotype. The results obtained may have implications for laboratory and clinical medicine, also in the light of recent findings suggesting that other mechanisms besides beta-lactamase hyperproduction may account for borderline susceptibility to PRPs. PMID- 8243484 TI - Plasmid carriage and antibiotic susceptibility of Neisseria meningitidis strains isolated in Sweden 1981-1990. AB - A random selection of Neisseria meningitidis strains isolated in Sweden in the period 1981-1990 were investigated for plasmid carriage and susceptibility to antimicrobial agents commonly used for treatment and prophylaxis of meningococcal disease. The MICs were determined by the agar dilution method for penicillin V, penicillin G, rifampicin, sulfadiazine, erythromycin and tetracycline. In 13% of the invasive strains the MIC of penicillin V was > or = 0.5 mg/l which may cause concern regarding the usefulness of penicillin V in prophylaxis. In strains isolated from the urogenital tract the MICs of penicillin V and penicillin G were higher than in the invasive strains. In about 82% of the strains isolated in 1987 1988 the MIC of tetracycline was > or = 0.5 mg/l whereas no such strains were found in 1981-1982. Plasmids were found in 2 of 119 invasive strains, in 1 of 50 strains from the respiratory tract and in 1 of 19 strains from the urogenital tract. The plasmid sizes were 1.3, 2.6, 25 and 40 Mda. None of these strains were beta-lactamase producing and no relation to a high degree antibiotic resistance was observed. PMID- 8243485 TI - Evaluation of pooled and individual components of Bordetella pertussis as antigens in an enzyme immunoassay for diagnosis of pertussis. AB - Six different antigen preparations for use in an enzyme immunoassay (EIA) to detect IgM, IgA and IgG antibodies to Bordetella pertussis were evaluated using sera from 13 randomly selected culture-positive patients and from 87 patients with suspected pertussis during a pertussis outbreak. Based on results in 80 healthy control sera a specificity limit of 99.9% was selected. Sera from all culture-positive patients reacted with at least one of the antigens. The sensitivity of the EIA using the individual antigen preparations was 85% for filamentous hemagglutinin, 92% for pertussis toxin, 62% for 69 kDa outer membrane protein, 85% for a pool of these three antigens, 54% for sonicated whole bacteria and 69% for 21 kDa pertussis toxin subunit S1. In the outbreak patient group 49 (56%) of the initial sera reacted with at least one of five antigen preparations. The EIA using sonicated bacteria detected only 41% of all seropositive cases compared with 51% using filamentous hemagglutinin, 61% using pertussis toxin, 65% using 69 kDa OMP and 65% using pooled antigen. It is concluded that either the pooled antigen or pertussis toxin antigen are suitable antigen preparations for use in the EIA for diagnosis of pertussis. PMID- 8243486 TI - Incidence of community-acquired pneumonia caused by Chlamydia pneumoniae in Italian patients. AB - The incidence of Chlamydia pneumoniae as a cause of community-acquired pneumonia was evaluated in a one-year prospective study in 108 patients with community acquired pneumonia. The bacteriological diagnosis was based on culture of sputum or bronchial aspirate and examination of acute and convalescent phase sera for Mycoplasma pneumoniae, Legionella pneumophila and Chlamydia pneumoniae. A definitive microbiological diagnosis was obtained in 58 (54%) patients. Chlamydia pneumoniae was the causative agent in 14 patients (13%) on the basis of positive serological tests; in 10 of the 14 patients Chlamydia pneumoniae was also detected by means of an indirect immunofluorescence test using pharyngeal swab specimens. In conclusion, Chlamydia pneumoniae seems to be a common etiological agent of community-acquired pneumonia, as increasingly reported in the last six to seven years. PMID- 8243487 TI - Parainfluenza virus infection in adult bone marrow transplant recipients. AB - The clinical course of parainfluenza virus infection occurring in 8 of 265 (3%) adult bone marrow transplant recipients during 1991 was reviewed. Parainfluenza virus type 3 was isolated from all eight patients. The clinical course ranged from self-limited upper respiratory tract infections (2 patients) to severe lower respiratory tract disease (6 patients) associated with a 50% mortality. This study highlights the important role of community respiratory viruses such as parainfluenza virus in the etiology of pneumonia in immunocompromised adults. PMID- 8243488 TI - Persistent human parvovirus B19 infection following an acute infection with meningitis in an immunocompetent patient. AB - A case in which parvovirus B19 infection persisted over a prolonged period of time in the blood of an immunocompetent patient following an acute infection with meningitis is reported. Using a nested polymerase chain reaction assay the viral genome was detected in cerebrospinal fluid as well as in blood at the time of overt disease and in consecutive blood samples collected for up to nine months. PMID- 8243489 TI - Three cases of Fusobacterium necrophorum septicemia. AB - Two cases of necrobacillosis or post-anginal septicemia (Lemierre's syndrome) due to Fusobacterium necrophorum are described. The first case in a young adult was complicated by thrombophlebitis of the internal jugular vein, lung abscesses and skin involvement. The second case was in a 2-year-old child, which is exceptional. The primary focus was possibly otitis media. In a third patient with multiple metastases of colon carcinoma, Fusobacterium necrophorum septicemia was diagnosed during a bout of fever 36 hours before death. PMID- 8243490 TI - Impact of cefpodoxime proxetil and amoxicillin on the normal oral and intestinal microflora. AB - Ten healthy volunteers were given 200 mg cefpodoxime proxetil tablets every 12 h and ten volunteers received 500 mg amoxicillin tablets every 8 h for seven days and the impact of the agents on the oral and intestinal microflora was studied. In the oral microflora, only minor alterations were observed in both groups. In subjects receiving cefpodoxime proxetil, the numbers of streptococci, enterobacteria and clostridia were strongly reduced in the faecal flora, while there was an overgrowth of enterococci, yeasts and Clostridium difficile. Amoxicillin administration induced somewhat smaller alterations in the faecal microflora, although all subjects had overgrowth of new colonizing amoxicillin resistant microorganisms, mainly Escherichia coli, Klebsiella and Enterobacter. Beta-lactamase activity was detected in the flora of six volunteers from each group. PMID- 8243491 TI - Fatal Clostridium perfringens meningitis associated with insertion of a ventriculo-peritoneal shunt. PMID- 8243492 TI - Isolation of Haemophilus paraphrophilus from an abdominal abscess. PMID- 8243493 TI - Development of pleural and pericardial effusions during itraconazole therapy of pulmonary aspergillosis. PMID- 8243494 TI - Fusidic acid induced jaundice. PMID- 8243495 TI - A new method to estimate parameters of pharmacokinetics with enterohepatic circulation. AB - A new method of estimating pharmacokinetic parameters for enterohepatic recirculation models is presented. The algorithm, based on the simplex method, assures a convergence toward the minimum minimorum; provides reliable parameters, even in large numbers; and handles up to five cycle effects in five established models: IV, EV, mono-, bi- or tricompartmental. For each model, the reabsorption rate may be considered to be slow (characterised by a rate constant with a significant value) or instantaneous (an infinite rate constant). Two examples are given to illustrate the qualities of the software that incorporates this new algorithm. The first relates to the pharmacokinetics of zinc orally given to humans; the second treats the kinetics of alpha-methylDOPA given to dogs intra arterially (note: values of plasma concentrations have been extracted from the literature). PMID- 8243496 TI - Metabolism and pharmacokinetics of [14C]-deoxyfructosylserotonin creatinine sulfate administered orally and intravenously to rats and mice. AB - Deoxyfructosylserotonin (DFS) has been shown in in vitro tests to inhibit L-DOPA oxidase and also to suppress the multiplication of Mycobacterium leprae. The possible therapeutic use of DFS makes necessary the study of its metabolic fate in animal models. Labelled [14C]-DFS was synthesized by condensation of serotonin and [14C]-glucose and administered per os or intravenously to rats and mice. After oral administration, some of the radioactivity transited through the intestinal tract to be excreted in feces (20-60% of the dose) and some was destroyed in the pH conditions of the intestine and further metabolized by the flora, producing 14CO2 in the expired air (10-40% of the dose). Radioactivity excreted in the urine amounted to 8-15% after 24 h. After intravenous administration, 60-90% of the dose had already been excreted in the urine after 8 h. Feces and CO2 accounted for 5-10% each. In the urine, for both routes of administration, beside DFS, half of the radioactivity corresponded to the glucuronide conjugate, while in the feces all the radioactivity found was unchanged DFS. Whole animal body autoradiography showed the presence of radioactivity in all the organs (1-2% of the dose) mainly resulting from the incorporation of labelled carbon from glucose and CO2. These results, obtained in healthy rats, demonstrate poor intestinal absorption of DFS (10% of the dose) and when it is absorbed, rapid urinary excretion. For its possible therapeutic use as an anti-leprosy drug in humans, derivatives with higher bioavailability must be attained. PMID- 8243497 TI - Pharmacokinetics of cutaneous Sulconazole nitrate in the hairless rat: absorption, excretion, tissue concentrations. AB - After cutaneous application of radioactive solutions of Sulconazole nitrate in the hairless rat, the total absorption of the substance by the skin, estimated from the sum of the cumulative urinary and fecal excretions over 96 h, was 2.4% of the dose administered. The elimination reached a maximum between 6 and 24 h and was virtually complete after 96 h. The excretion was almost equally distributed between the urine and the feces, which corresponds to an intense elimination via the biliary tract. The quantities present in the stratum corneum, epidermis and dermis at the end of the period of contact constituted another estimation of the total absorption of the substance which confirmed the previous estimation (3.6% of the dose). The measurement of the concentrations of Sulconazole and its metabolites in the various layers of the skin revealed a high affinity of the substance for the stratum corneum, where it remained present in large quantities for more than 48 h. This affinity is due to the very intense lipophilia of the molecule. The concentrations in the other tissues were inversely proportional to the distance from the surface of the skin and were virtually nil in the circulating blood. These results suggest the absence of risk of systemic effects after cutaneous administration of Sulconazole and support the recommended therapeutic protocol in man (one administration per day). PMID- 8243498 TI - Aspects of schizandrin metabolism in vitro and in vivo. AB - The metabolic transformation of schizandrin, isolated from the kernel of Schizandra chinensis Bill, was studied in vitro with a phenobarbital induced rat liver microsomal fraction containing the NADPH generating system. The major metabolites were isolated by preparative HPLC and identified as 7,8-dihydroxy schizandrin, 7,8-dihydroxy-2-demethyl-schizandrin, and 7,8-dihydroxy-3-demethyl schizandrin by UV, NMR and MS spectral analysis. The 7,8-dihydroxy-schizandrin was confirmed further by comparison with spectral and chromatographic behavior of the authentic compound. The metabolic biotransformation of schizandrin in vivo was also determined. PMID- 8243499 TI - Pharmacokinetics of d(+)-usnic acid in rabbits after intravenous administration. AB - The pharmacokinetics of usnic acid, a lichen antitubercular, antitumor and enzyme inhibiting agent, was studied in normal male rabbits after administration of 5 mg/kg dose by intravenous route. Plasma samples were collected up to 48 h. The plasma usnic acid levels showed a triexponential decay with a terminal half life of 10.69 h. The mean volume of distribution of the central compartment was 43.93 ml/kg. The pharmacokinetic parameters obtained from a three-compartment body model and a noncompartment body model gave close values. The mean steady-state volume of distribution was 167.17 and 163.29 ml/kg, systemic clearance was 12.17 and 12.25 ml/h/kg and AUC was 428.51 and 426.69 micrograms.ml/h following three compartmental and noncompartmental analyses respectively. PMID- 8243500 TI - Effect of induced hypoalbuminemia on distribution, total clearance and unbound clearance of piroxicam in vivo in the rat. AB - The influence of decreased albumin concentration on the pharmacokinetic behaviour of Piroxicam was studied in vivo in rats that had undergone plasmapheresis. Reductions of approximately 25% and 50% in the plasma albumin concentration were achieved, the former in rats not given plasma expander, the latter in animals given Ficol-70 as a plasma expander. The unbound fraction of Piroxicam in plasma and the apparent volume of distribution at steady state experienced a statistically significant increase where the albumin concentration was reduced. The average total plasma clearance rose with the increase in fu, between the control (6.3 +/- 2.4 ml/h) and plasmapheretic groups (11.1 +/- 4 ml/h), in accordance with predictions of the 'well-stirred' and 'parallel-tube' models, but no statistically significant differences were found between the two groups, perhaps because of the great interindividual variability associated with this parameter. The total plasma clearance value of the Ficol group (5.4 +/- 2.2 ml/h) was close to that of the control group, despite the high increase in the unbound fraction in plasma. Alterations in the uptake process in the liver due to the high level of induced hypoalbuminemia may have occurred. PMID- 8243501 TI - Absorption, tissue distribution and excretion of radiolabelled compounds in rats after administration of [14C]-L-alpha-glycerylphosphorylcholine. AB - The kinetics and metabolism of L-alpha-glycerylphosphoryl-choline (alpha-GPC) were investigated in male and female rats after i.v. (10 mg/kg) and oral doses (100-300 mg/kg). alpha-GPC was labelled with [14C]-glycerol ([14G]-GPC) or [14C] choline ([14C]-GPC). Different kinetic and metabolic profiles were observed after i.v. and oral administration. It is assumed that alpha-GPC is hydrolyzed by phosphodiesterases in the gut mucosa. The different labelled metabolites have different kinetic properties of absorption, distribution and clearance, leading to different blood concentration-time curves of total radioactivity. Both labelled compounds gave a wide distribution of radioactivity, particularly concentrated in the liver, kidney, lung and spleen compared to blood. Brain concentrations of [14C]-GPC were comparable to ([14G]-GPC) or lower than ([14C] GPC) total blood radioactivity. The metabolite profile in the perfused brain showed a small amount of choline and two unknown metabolites, probably the same as in blood. In addition, choline was incorporated into brain phospholipids in increasing amounts within 24 h of dosing. In all cases renal and fecal excretion of radioactivity was low and comparable for [14G]-GPC and [14C]-GPC. Mostly the administered radioactivity was exhaled as 14CO2, this degradation being faster and more pronounced for the glycerol-labelled metabolites than for the choline labelled metabolites for both routes of administration. In all cases the results were the same for male and female rats. PMID- 8243502 TI - Effect of pregnandiol on caffeine metabolism in female rats. AB - Three groups of six 5-week-old Sprague Dawley female rats received i.p. injections of pregnandiol, 1.25, 2.50 or 5 mg/kg, respectively, in triolein daily for 7 days. Caffeine metabolism was studied in liver slices on day 8 by HPLC. Only primary metabolites were formed. N-1 demethylation was the most important pathway (theobromine represented 51% of total dimethylxanthines). Unlike in human in vitro or in vivo, 1,3,7-DAU (6-amino-5-(N-formylmethylamino)-1,3 dimethyluracil) was an important metabolite (9.7% of total caffeine metabolites). Pregnandiol inhibited N-1, N-3 and N-7 demethylation in vitro (-33%, -33% and 28%, respectively, at 5 mg/kg/day), but it had no effect on N-1 demethylation at 1.25 or 2.50 mg/kg/day. Pregnandiol at all doses had no effect on 1,3,7 trimethyluric acid and 1,3,7-DAU formation. These results are consistent with the hypothesis that C-8 hydroxylation and demethylation of caffeine are mediated by different isoenzymes. They indicate that pregnandiol is a potent inhibitor of microsomal drug metabolism, specifically of cytochrome P450 IA, which could explain the immaturity of some metabolic pathways of caffeine in neonates. PMID- 8243503 TI - Metabolism of S-nicotine in noninduced and aroclor-induced rats. AB - The urinary excretion of nicotine and its metabolites in noninduced and Aroclor induced male and female rats has been determined following intravenous administration of 2'-[14C]-labeled S-nicotine at a dose of 4.6 mumol/kg. Complete recovery of the administered radioactivity was achieved: 95% in urine and 4% in feces over 96 h and 1% remaining in the body. More than 40 nicotine metabolites were found by radio-HPLC; 19 were identified including the cis/trans diastereomers of nicotine-N'-oxide and 3'-hydroxycotinine. The urinary metabolite profile and excretion kinetics of nicotine and its metabolites were significantly different between noninduced and Aroclor-induced rats. The major urinary nicotine metabolite in the noninduced rat was cis-nicotine-N'-oxide. In the Aroclor induced rat, cotinine metabolites were the major metabolites found. Sex differences were found for the urinary nicotine metabolite profile, mainly expressed in the excretion of cis-nicotine-N'-oxide, 29% in the male and 17% in the female noninduced rat, and the excretion of cotinine, 5% in the male and 12% in the female noninduced rat. High stereoselectivity was found for the formation of the cis/trans-diastereomers of nicotine-N'-oxide as well as of 3' hydroxycotinine, the stereoselectivity being more pronounced in male rats. PMID- 8243504 TI - Pharmacokinetics and metabolism of diltiazem in healthy males and females following a single oral dose. AB - Plasma concentrations and urinary excretion of DTZ and its metabolites were determined in 20 healthy volunteers (10 males and 10 females) after they had each been given a single oral 90 mg dose of DTZ. DTZ and six of its metabolites which included N-monodesmethyl DTZ (MA), deacetyl DTZ (M1), deacetyl N-monodesmethyl DTZ (M2), deacetyl O-desmethyl DTZ (M4) and deacetyl DTZ N-oxide (M1NO) and deacetyl N,O-didesmethyl DTZ (M6), were determined by a sensitive and specific HPLC assay. The major metabolites measurable in the plasma of all the volunteers were MA, M1, and M2. The terminal half-lives (t1/2) of M1 and M2 were considerably longer than those of DTZ and MA. Less than 5% of the dose was excreted as unchanged DTZ in the urine over the 24 h period. The major urinary metabolite was MA, followed by M6, M2, and then M1. Except for the urinary excretion of M4 there were no statistically significant differences in any of the pharmacokinetic parameters between the males and the females. The mean 24 h urinary recovery of M4 was higher in the males than in the females (P < 0.05). However there were large inter-individual variations in the plasma concentrations and urinary excretion of DTZ and its metabolites with some parameters differing by more than 20-fold. In addition, O-desmethyl DTZ (Mx) and N,O-didesmethyl DTZ (MB) were identified as two other major urinary metabolites. PMID- 8243505 TI - Digitoxin and its metabolites in patients with liver cirrhosis. AB - Pharmacokinetic profile and urinary excretion of digitoxin and 4 metabolites were investigated in 9 patients with biopsy confirmed liver cirrhosis (median antipyrine clearance 20.0 +/- 5.4 ml/min; X +/- SEM) and were compared with that of 8 healthy volunteers (antipyrine clearance 36.9 +/- 4.9 ml/min) following intravenous and p.o. administration of 1 mg digitoxin. The kinetic parameters derived from the digotoxin plasma concentration time curve and from urinary recovery including total clearance of unchanged digitoxin did not differ significantly between both groups investigated. Renal clearance of digitoxin was 0.017 +/- 0.005 ml/min/kg in the patient group and 0.011 +/- 0.002 ml/min/kg in the volunteers (NS); it was 0.00340 +/- 0.00047 ml/min/kg and 0.00223 +/- 0.00039 ml/min/kg, respectively for digitoxigenin-bis-digitoxoside (NS), 0.00006 +/- 0.00001 ml/min/kg and 0.00016 +/- 0.00005 ml/min/kg for digitoxigenin-mono digitoxoside (P < 0.05), 0.00041 +/- 0.00013 ml/min/kg and 0.00088 +/- 0.00032 ml/min/kg for digitoxigenin (P < 0.05), 0.00135 +/- 0.00049 ml/min/kg and 0.00113 +/- 0.00042 ml/min/kg for digoxin (NS). In conclusion, hydrolysis of digitoxin is altered in liver cirrhosis, whereby a significant reduction in the renal clearance and urinary recovery of digitoxigenin-mono-digitoxoside and digitoxigenin was seen in the present study. PMID- 8243506 TI - Some pharmacokinetic considerations about homeostatic equilibrium of endogenous substances. AB - Endogenous substances in the body are controlled through simple, very effective mechanisms, that preserve an optimum homeostatic equilibrium of baseline concentration and restore it when impaired. When planning a pharmacokinetic investigation of an endogenous substance exogenously administered, it is imperative to carefully ascertain the above mechanisms as well as the baseline value and their possible variations associated with daily rhythm, food, age, sex, menstrual cycle. Often the control mechanisms operate through non-linear processes, therefore a non-compartmental analysis or a tailored model may be more appropriate than the compartmental models used in standard pharmacokinetic analysis. Some specific examples of endogenous substances are discussed here on the basis of the data from the literature and personal experience. PMID- 8243507 TI - Management of acute peripheral vestibular disorders. PMID- 8243508 TI - Genetic contribution to idiopathic adult-onset blepharospasm and cranial-cervical dystonia. AB - A family study in 29 patients with idiopathic adult-onset blepharospasm (n = 16) and cranial-cervical dystonia (n = 13) was undertaken by examining 189 first degree relatives. Six relatives with dystonia were identified in 6 families. A further 3 affected relatives, now deceased, were from 2 other families. All the secondary cases were parents or siblings. There was a tendency for affected relatives to have the same type of dystonia of index patients. To assess the significance of secondary cases we compare the incidence of affected siblings between probands and their spouses. Dystonia was found in 5 out of 120 proband siblings whereas none of the 142 spouse siblings was affected (p < 0.05). Segregation analysis suggested an autosomal-dominant transmission and reduced penetrance or, alternatively, polygenic inheritance. PMID- 8243509 TI - Cognitive behavior in heredodegenerative ataxias. AB - Fifteen patients with Friedreich's ataxia (FA) and 15 others with olivopontocerebellar atrophy (OPCA) were evaluated with a comprehensive neuropsychological battery of tests. They were pair-matched with normal controls for age, sex and education. Depressed patients were excluded from the study as were those with extrapyramidal signs. The following results were obtained: (1) the Raven test, untimed block design performance in OPCA and quantitative analysis of Rey figure drawing revealed a visuospatial deficit suggestive of a mild parietal-like syndrome; signs of mild frontal-like syndrome were also found; (2) cognition was slowed in the FA group; (3) simple visual and auditory reaction times were increased in both patient groups. It appears that the cerebellum may interfere indirectly with cognition through various physiological and neurochemical 2-way cerebellocortical loops. Finally, the cerebellum seems to interfere directly also with basic speed of information processing. PMID- 8243510 TI - Involvement of cytokines in N-methyl-N'-nitro-N-nitrosoguanidine-induced plasminogen activator activity in acute disseminated encephalomyelitis and multiple sclerosis lymphocytes. AB - In order to examine the plasminogen activator (PA) induction involved in the pathogenesis of acute disseminated encephalomyelitis (ADEM) and multiple sclerosis (MS), PA activity in peripheral blood lymphocytes derived from 5 ADEM and 3 MS patients was investigated. There was no PA induction in any ADEM, MS or control lymphocytes treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) alone. PA activity, however, in lymphocytes exposed to human interferon-gamma (HuIFN-gamma) prior to MNNG treatment was elevated during the active phase of ADEM and MS, whereas the PA induction disappeared in association with improvement of the neurologic symptoms. The PA activity was abolished by mixed treatment with HuIFN-gamma and anti-HuIFN-gamma antibody. No such PA induction by any HuIFN was observed in any normal controls or cases of other neurologic diseases. Among the cytokines tested other than HuIFN, tumor necrosis factor-alpha, in combination with MNNG, also induced PA activity in lymphocytes from ADEM and MS patients during the active phase. Thus, the PA induction observed in lymphocytes on combined treatment with MNNG and cytokines may be involved in the progression of neurologic disorders in these demyelinating diseases, and indicates the possibility of therapeutic strategies involving anticytokine usage. PMID- 8243511 TI - Assessment of sudomotor dysfunction in early Parkinson's disease. AB - We performed a quantitative sudomotor function test on 10 patients with early Parkinson's disease (PD) and 11 age-matched control subjects. Thermal warming increased the sweat rate in both PD and controls. There was no difference in sweat rate between the forearm and the thigh in either PD or controls. In PD, thyrotropin-releasing hormone (TRH) did not increase the sweat rate, whereas it did so in the controls. These results suggest that sudomotor dysfunction in early PD is minor but that there may be an impairment of TRH-induced sympathetic response in the early stages of PD. PMID- 8243512 TI - Diffuse cerebral hypoperfusion in epileptic patients observed from quantitative assessment with single photon emission computed tomography using N-isopropyl (iodine-123)-p-iodoamphetamine. AB - Quantitative assessment of regional cerebral blood flow (rCBF) in the bilateral cerebral and cerebellar cortices was performed in 15 epileptic adult patients receiving chronic high-dose antiepileptic drug therapy and 22 normal volunteers matched for sex and age, using single photon emission computed tomography with N isopropyl-(iodine-123)-p-iodoamphetamine. The entire averaged rCBF value in the epileptic patients, i.e. 52.8 +/- 13.7 ml/10 g/min (range: 25-78 ml/100 g/min), was significantly lower as compared with that in the normal subjects, i.e. 69.1 +/- 14.2 ml/100 g/min (range: 46-102 ml/100 g/min). Six of the 15 patients showed absolute rCBF values less than the minimum of the normal range, i.e. 46 ml/100 g/min, in all or most of the measured brain tissues. There was a significant correlation between the diffuse cerebral hypoperfusion and simultaneous ingestion of phenytoin and phenobarbital. The possible effects of antiepileptic drugs on rCBF are discussed. PMID- 8243513 TI - Auditory event-related potentials in patients with systemic lupus erythematosus. AB - We studied auditory event-related potentials (ERPs) in 17 patients with systemic lupus erythematosus (SLE). Latencies of N100 and P200 components were normal in all patients, whereas the latencies of target P300 and nontarget P300 components were significantly prolonged in SLE patients with and without cognitive dysfunction. The P300 component of ERPs can be applied to evaluate the cognitive aspects of central nervous system manifestations in SLE patients. PMID- 8243514 TI - Spontaneous collapse of posttraumatic syringomyelia: serial magnetic resonance imaging. AB - The case of a 30-year-old man with posttraumatic syringomyelia involving the cervical and upper thoracic spine is reported. The patient was followed clinically and with magnetic resonance (MR) imaging over a 3-year period. Spontaneous complete remission of the syrinx-related symptoms correlated with a partial collapse of the syrinx, as proved by serial MR imaging. PMID- 8243515 TI - Unilateral trismus caused by vertebrobasilar dolichoectasia. AB - Vertebrobasilar dolichoectasia is a relatively uncommon pathological entity and generally asymptomatic. We report a quite unusual case of unilateral motor trigeminal involvement with trismus, due to VD. Apart from tetanus or local morbid conditions of the mouth, trismus is often attributed to disturbed programming and co-ordination of the masticatory muscles within the mesencephalic nucleus. The possibility of truncal compression of the trigeminal motor root by vertebrobasilar dolichoectasia being responsible for masseter muscle spasm, in analogy with the pathogenetic mechanisms proposed in hemifacial spasm, is proposed as an alternative explanation in the present case. PMID- 8243516 TI - Reversible motor neuron disease. AB - We investigated a 69-year-old male with a clinical syndrome resembling amyotrophic lateral sclerosis characterized by fasciculation, wasting of the limb muscles and increased deep tendon reflexes in the lower limbs. Electromyographic (EMG) studies showed abundant positive sharp waves and fibrillation potentials with decreased recruitment in the limbs and paraspinal muscles. The patient recovered almost completely in approximately 1.5 years, and follow-up EMG studies showed no positive sharp waves or fibrillation potentials in the limb muscles except for some polyphasic motor units in the bilateral intrinsic hand muscles. No known systemic disease, malignancy or heavy metal intoxication was found during the course of his illness. So far, there are only few cases reported with spontaneous remission of motor neuron disease; however, the possibility should always be considered. PMID- 8243517 TI - Multiple sclerosis-like syndrome in a woman heterozygous for adrenoleukodystrophy. AB - A 28-year-old asymptomatic woman was diagnosed to be heterozygous for adrenoleukodystrophy (ALD) by elevated very long-chain fatty acids in serum and fibroblasts after ADL had been diagnosed in her son. A year later she had transient unilateral blurred vision. Evoked potentials and brain magnetic resonance imaging showed further separate cerebral white matter lesions suggesting multiple sclerosis (MS). MS-like syndromes in women heterozygous for ALD may be more frequent than previously recognized. PMID- 8243518 TI - Computed tomography and magnetic resonance imaging in three patients with Tolosa Hunt syndrome. AB - Three patients with Tolosa-Hunt syndrome were examined by computed tomography (CT) and magnetic resonance imaging (MRI). CT of the brain did not reveal any definite abnormality in any of the patients. MRI demonstrated an abnormal soft tissue area in the cavernous sinus in only 1 patient. There was clinical improvement after corticosteroid therapy. MRI is mandatory for demonstrating lesions in the cavernous sinus or in the superior orbital fissure in patients with THS. PMID- 8243519 TI - Pachymeningeal thickening in rheumatoid arthritis. PMID- 8243520 TI - Hereditary motor and sensory neuropathy type V with spinal cord atrophy on magnetic resonance imaging. PMID- 8243521 TI - The platelet angiotensin II receptor in type I diabetes: studies in patients with and without nephropathy. AB - Experimental studies demonstrate impaired regulation of the mesangial angiotensin II (AII) receptor in diabetes. This could contribute to the disturbance of glomerular blood flow and the development of diabetic nephropathy. The aim of this study was to determine whether a similar receptor abnormality occurs in patients with type I insulin-dependent diabetes mellitus (IDDM) and if so whether this is more prevalent in patients with micro- or macro-albuminuria. The platelet AII receptor was chosen because of its availability from the circulation and its comparable regulatory properties to tissue-based receptors. The interaction between plasma AII and its platelet receptor was examined in 45 patients with IDDM and 36 age- and sex-matched control subjects. Seven patients had clinical nephropathy and two had persistent micro-albuminuria. The duration of diabetes varied from 1 month to 42 years. There was a significant inverse correlation between plasma AII and the logarithm of receptor number in the control group (r = -0.555, P < 0.001). This relationship was not observed in the diabetic patients irrespective of the duration of disease or the presence of nephropathy. Receptor expression in patients without nephropathy showed no correlation with either duration of disease or the degree of glycaemic control. However, a significant relationship between AII receptor number and duration of diabetes was noted in the group with nephropathy (r = 0.723, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243522 TI - Effect of duodenal juice on bombesin-stimulated cholecystokinin release during loxiglumide administration in man. AB - Stimulation of cholecystokinin release by bombesin in augmented by cholecystokinin receptor blockade with loxiglumide. We hypothesize that this augmented cholecystokinin release results from inhibition of the pancreatico biliary response to bombesin during cholecystokinin receptor blockade. To test this hypothesis, we infused bombesin for 180 min in six healthy subjects Three bombesin-infusion experiments were performed in each subject in random order on different days. In two of these experiments loxiglumide was co-infused with bombesin, while in the third experiment saline was co-infused with bombesin. In one of the loxiglumide experiments, duodenal juice, collected on the previous day during infusion of cholecystokinin-GIH, was reperfused intraduodenally during the second hour of bombesin infusion. In the saline experiment, the integrated cholecystokinin response during the first hour of bombesin-infusion (262 +/- 63 pmol 60 min-1) was significantly (P < 0.01) higher than during the second (88 +/- 26 pmol 60 min-1) and third (87 +/- 31 pmol 60 min-1) hour of bombesin-infusion. Loxiglumide augmented bombesin-stimulated cholecystokinin secretion from 262 +/- 63 pmol 60 min-1 to 453 +/- 63 pmol 60 min-1 in the first hour of bombesin infusion (P < 0.01). Integrated cholecystokinin values in the second (489 +/- 90 pmol 60 min-1) and third (450 +/- 74 pmol 60 min-1) hour of the loxiglumide experiment, were significantly (P < 0.01) higher than in the saline experiment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243523 TI - Composition of ascitic fluid in liver cirrhosis: bile acid and lipid content. AB - The concentrations of lipids, bile acids and proteins were evaluated in the ascitic fluid and plasma of 23 cirrhotics. Ascitic fluid density was highly correlated with its protein content, represented mostly by low molecular weight proteins. The ratio of plasma to ascitic fluid concentrations of nine examined proteins increased with molecular weight, indicating a selective ultrafiltration of the peritoneal transudate. Low density lipoproteins in ascitic fluid had modified electrophoretic mobility. Total cholesterol had a higher plasma to ascitic fluid ratio than high density lipoprotein cholesterol, whereas bile acids and proteins had similar plasma to ascitic fluid ratios. Indeed, bile acids strongly bind to circulating albumin: consequently ascitic fluid contains more cholic acid (less hydrophobic) than other bile acids. Analysis of both plasma and ascitic fluid composition in cirrhotics provides useful information on processes regulating passage of blood components into the peritoneal cavity. PMID- 8243524 TI - Effect of IgM-enriched intravenous immunoglobulin (Pentaglobin) on endotoxaemia and anti-endotoxin antibodies in bone marrow transplantation. AB - Endotoxin was measured in over 1000 plasma samples from bone marrow transplant patients in a randomized trial of the IgM-enriched intravenous immunoglobulin (IVIG) Pentaglobin. Peak endotoxaemia was significantly reduced (P = 0.02) in patients receiving Pentaglobin and 70% of all pyrexias of unknown origin were associated with endotoxaemia. Gut mucosal damage, assessed by lactulose/mannitol ratios, was significantly associated (P = 0.02) with endotoxaemia. Specific IgM antibody to endotoxin core-glycolipid was significantly raised (P < 0.01) in patients receiving the IVIG, and the IgM fraction of Pentaglobin was found to contain most of the anti-endotoxin antibody activity of the IVIG. These results suggest a role for IgM-enriched IVIG as a prophylactic agent for the reduction of endotoxaemia and its consequences in bone marrow transplant patients. PMID- 8243525 TI - Cica-clamp technique: a method for quantifying parathyroid hormone secretion: a sequential citrate and calcium clamp study. AB - Objectives of the present study were to establish and investigate a standardized method for quantifying of intact parathyroid hormone secretion during sequential induction of hypo- and hypercalcaemia, and to explore the applicability to these data of a mathematical model derived from in vitro studies as presented in the literature. Twenty-two healthy volunteers aged 20-80 years participated in one or more experiments. The experiments comprised three different protocols of sequential induction of a regular hypocalcaemic (citrate) clamp followed by increases in blood ionized calcium, ending in a regular hypercalcaemic (calcium) clamp. During protocol I, the induction of hypocalcaemia, blood ionized calcium 0.21 mmol l-1 (SD 0.01, n = 76) below baseline, the release of serum parathyroid hormone rapidly increased to a concentration of four to seven times above baseline. The serum parathyroid hormone declined gradually to a steady state of about two to three times above baseline. During stepwise increases in blood ionized calcium, the serum parathyroid hormone rapidly declined to new steady state concentrations. When a hypercalcaemia of 0.20 mmol l-1 (SD 0.02, n = 76) above baseline was reached, serum parathyroid hormone was suppressed to about one fourth of baseline concentration. Protocol II, the Cica-clamp, and protocol III, are short versions of protocol I using a slow and gradual increase in blood ionized calcium from hypo- to hypercalcaemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243526 TI - Plasma aluminium in a reference population: the effects of antacid consumption and its influence on biochemical indices of bone formation. AB - Aluminium is involved in the etiology of several complications of chronic renal failure and has been firmly established as having toxic effects on bone tissue. We have measured plasma aluminium together with serum osteocalcin, procollagen I C-terminal peptide and total alkaline phosphatase activity in healthy subjects and in a group of subjects who consumed aluminium-containing and non-aluminium containing antacid preparations, with normal renal function. Age-related healthy reference ranges for plasma aluminium are presented and the effects of chronic antacid consumption on plasma aluminium and biochemical markers of bone formation investigated. In 172 healthy subjects the mean plasma aluminium concentration was 4.4 +/- 2.9 micrograms L-1, men having a significantly greater circulating aluminium load than women (5.4 +/- 2.8 micrograms L-1 vs. 4.0 +/- 2.8 micrograms L-1 respectively (P = 0.0039)). Older men were found to have significantly higher plasma aluminium levels than younger men. Increased plasma aluminium was seen in subjects taking antacids although this was not associated with significant changes in most indices of bone formation. PMID- 8243527 TI - In vivo kinetics of lipoprotein(a) in homozygous Watanabe heritable hyperlipidaemic rabbits. AB - In vivo kinetics of lipoprotein(a) [Lp(a)] were investigated in homozygous Watanabe heritable hyperlipidaemic (WHHL) rabbits (an animal model of familial hypercholesterolemia (FH)), and in normolipidemic Japanese White rabbits (controls). 125I-labelled Lp(a) and 131I-labelled LDL were simultaneously injected intravenously. Blood samples were then taken periodically. Kinetic parameters were calculated from the plasma radioactivity decay curves. The fractional catabolic rates (FCRs) of both Lp(a) and LDL (1.355 +/- 0.189 pools per day and 1.278 +/- 0.397 pools per day, respectively) in the WHHL rabbits were significantly (P < 0.005) smaller than those in the control rabbits (2.008 +/- 0.083 pools per day and 2.855 +/- 0.759 pools per day, respectively). In WHHL rabbits, the FCRs of Lp(a) and LDL were similar. However, in control rabbits, the FCR of Lp(a) was significantly (P < 0.01) smaller than that of LDL. In WHHL rabbit organs, the mean ratio of 125I-Lp(a): 131I-LDL, 48 h after injection, normalized to the corresponding isotope ratio in plasma, were 1.525, 1.020, 1.819 and 1.967, in liver, kidney, spleen and bile, respectively. These values were significantly higher than the corresponding values in control rabbits (0.590, 0.677, 0.862 and 0.766, respectively). Our data strongly suggest that Lp(a) clearance is not entirely dependent upon LDL receptors and may be mediated by some other mechanisms. PMID- 8243528 TI - Biochemical markers of bone turnover in women with surgically treated carcinoma of the breast. AB - Biochemical markers of bone turnover were measured in fasting urine and blood samples obtained from 38 postmenopausal women with previous surgical treatment of breast cancer combined with adjuvant chemotherapy, tamoxifen, or placebo. Significantly elevated urinary pyridinoline as nmol mmol-1 creatinine (47.5 and 42.5 in tamoxifen and placebo treated patients compared with 26.3 in normal controls, both P < 0.001) and deoxypyridinoline (11.9 and 10.5 compared with 6.3, P < 0.001 and P = 0.002 respectively) were found with unchanged urinary hydroxyproline, serum alkaline phosphatase and procollagen I carboxyterminal peptide (PICP). These findings suggest enhanced bone resorption resulting from the humoral osteoclast activating effect of the previous breast cancer or underlying carcinoma recurrence. Alternatively the raised pyridinium excretion might indicate an altered crosslinking composition of bone collagen. No specific effect on bone metabolism was found with tamoxifen treatment as all measured parameters were similar in both tamoxifen ex-users and non-users. This confirmed the safety of tamoxifen therapy with respect to bone. PMID- 8243529 TI - The influence of pretransplant lipoprotein abnormalities on the early results of renal transplantation. AB - Lipoprotein patterns were investigated before and after renal transplantation in a prospective study including 151 patients. Kidney graft losses during the first 6 months were associated with higher total cholesterol (P = 0.03), LDL cholesterol (P = 0.003) and LDL triglyceride levels (P = 0.01) before transplantation. Patients with serum cholesterol > or = 6.9 mmol l-1 before transplantation had more acute rejections (1.7 vs. 0.9), a worse graft function and more vascular intimal hyperplasia and glomerular mesangial changes in transplant biopsies at 6 months. Patients with serum creatinine levels exceeding 160 mumol l-1 at 6 months had more severe lipid disorders already before transplantation. Serum creatinine at 6 months was influenced by the number of acute rejection episodes (P = 0.0001) and the age of the donor (P = 0.009) while the number of acute rejections was found to be related to pretransplant total cholesterol levels (P = 0.0086) and the age of the recipient (P = 0.025). In conclusion, pretransplant lipoprotein disturbances have an impact on the early outcome of renal transplantation. Since there is a progression of hyperlipidaemia following transplantation, this may have an influence also on the cardiovascular morbidity and late graft dysfunction. PMID- 8243530 TI - Lipoprotein(a) levels in relation to diabetic complications in patients with non insulin-dependent diabetes. AB - The relationship between serum levels of lipoprotein(a) Lp(a)) and the presence of chronic diabetic complications was studied in 194 patients with non-insulin dependent diabetes mellitus (NIDDM; 75 males, 119 females; age 66 +/- 11 years; duration of diabetes, 11 (range 1-35) years). They were taking various treatments (diet alone, oral hypoglycaemic agents and/or insulin). Metabolic status and prevalence of diabetic complications were assessed by detailed history, physical examination, laboratory analysis and ECG. Average metabolic control was moderate (HbA1c 8.2 +/- 1.7%). Median serum Lp(a) level was 183 U l-1 (range 8-2600 U l 1), which was significantly higher than in control subjects of comparable age (median 101; range 8-1747 U l-1; P < 0.05), while HDL-cholesterol levels were lower (1.14 +/- 0.38 vs. 1.35 +/- 0.35 mmol l-1; P = 0.001), and total cholesterol levels were comparable. No significant relationships between diabetes treatment or metabolic control and Lp(a) levels were observed. In the quartile of patients with the highest Lp(a) levels, total cholesterol and triglycerides were slightly higher (P < 0.05), whereas HDL-cholesterol was not different. With increasing Lp(a) levels, higher prevalences of preproliferative retinopathy and of coronary artery disease (CAD) were observed, but not of the other complications. No relationship was found between the degree of albuminuria and Lp(a) levels. We conclude that in NIDDM patients, Lp(a) levels are elevated compared with non-diabetic subjects, and that higher Lp(a) levels are associated with higher prevalences of CAD and of retinopathy. PMID- 8243531 TI - Forearm muscle metabolism in primary hypothyroidism. AB - Thyrotoxicosis is associated with increased skeletal muscle glucose metabolism and lipid oxidation in the fasted state. Conversely, in hypothyroidism, glucose metabolism is reduced. The present study has assessed in patients the effects of chronic thyroid hormone deficiency, on insulin-stimulated skeletal muscle metabolism, using a combination of the forearm and hyperinsulinaemic euglycaemic clamp techniques. Seven female patients presenting with primary auto-immune hypothyroidism were studied at presentation and 4-9 months after being rendered biochemically euthyroid. No significant differences were demonstrated between the hypothyroid and euthyroid states in forearm muscle blood flow, or in glucose or lipid metabolism. PMID- 8243532 TI - Big-endothelin-1 contracts rat isolated uterus via a phosphoramidon-sensitive endothelin ETA receptor-mediated mechanism. AB - The presence of a phosphoramidon-sensitive endothelin-1-converting enzyme was investigated in the rat isolated uterus. Endothelin-1 and its precursor, big endothelin-1, increased the rate of spontaneous contractions and caused tonic contractions. Responses to big-endothelin-1 had a slower start than those to endothelin-1. The tonic contraction induced by big-endothelin-1 (10 nM) was nearly abolished by phosphoramidon (100 microM), but the response to an equieffective concentration of endothelin-1 (3 nM) was not affected. Big endothelin-1 (EC50 6.7 nM) was only 7-fold less potent than endothelin-1 (EC50 0.9 nM), whereas endothelin-3 was much less potent (EC50 > 100 nM). The endothelin ETA receptor antagonist, BQ-123 (40, 150 and 600 nM), induced graded rightward shifts of the concentration-response curve for endothelin-1. Schild analysis yielded a straight line with a slope not different from unity, and a pA2 value of 7.76. At 100 nM, BQ-123 specifically blocked responses to both endothelin-1 (3 nM) and big-endothelin-1 (10 nM), without modifying those to oxytocin (5 nM), acetylcholine (3 microM) or bradykinin (0.5 nM). Our results suggest the presence of phosphoramidon-sensitive endothelin-converting enzyme and demonstrate the occurrence of functional endothelin ETA receptors in the rat uterus. PMID- 8243533 TI - Ionic basis for OPC-8212-induced increase in action potential duration in isolated rabbit, guinea pig and human ventricular myocytes. AB - Changes in transmembrane ionic currents induced by OPC-8212 (3,4-dihydro-6-[4 (3,4-dimethoxybenzoyl)-1-piperazinyl]-2(1H)-quinoline) , a recently introduced positive inotropic agent which lengthens cardiac action potential duration, were examined using whole-cell voltage-clamp techniques in single rabbit, guinea pig and human ventricular myocytes. In rabbit, OPC-8212 (12 mumol/l) significantly increased membrane action potential duration measured at 90% of repolarization by an average of 88 ms (from 462 +/- 25 to 550 +/- 35 ms, n = 4; P < 0.05). In rabbit this increase in duration was not associated with significant changes in either the inward rectifier or transient outward K+ currents. The magnitude of the secondary inward current evoked from a holding potential of -50 mV was significantly increased by 97 +/- 8% (n = 6; P < 0.01) while a demonstrable delayed rectifier outward current could not be identified in the rabbit myocytes examined at room temperature. In guinea pig ventricular myocytes, where the delayed rectifier was large, 12 mumol/l OPC-8212 significantly depressed the current by 58 +/- 10% (n = 6; P < 0.01). The effects of OPC-8212 in human ventricular myocytes obtained from the explanted heart of a single patient having an idiopathic cardiomyopathy most closely resembled those observed in isolated rabbit ventricular myocytes. Thus, in rabbit and a few human ventricular myocytes examined at room temperature, OPC-8212 appeared to lengthen cardiac membrane action potential duration primarily by increasing the amplitude of the secondary inward current believed to primarily represent current through L-type Ca2+ channels. In guinea pig preparations, OPC-8212 also decreased the delayed rectifier outward K+ current which also would account for an increase in action potential duration. OPC-8212 could not be demonstrated to affect Na+ current inactivation in a manner similar to that produced by 1 mg/l veratrine, a recognized Na+ channel agonist, which dramatically slowed this process. PMID- 8243534 TI - Antagonist effect of losartan on angiotensin II induced contraction in five isolated smooth muscle assays. AB - Antagonist effect of losartan, a nonpeptide antagonist of angiotensin II, on angiotensin II induced contractile response was studied in five isolated smooth muscle assays. In the rabbit aorta and guinea-pig stomach assays, losartan competed with angiotensin II for the angiotensin receptors in an apparently simple manner, that is compliance with the basic criteria of Schild analysis for simple competition. Noncompliance, however, was observed in the guinea-pig ileum, rat ileum and guinea-pig trachea assays where losartan induced nonparallel rightward shifts of angiotensin II E/log[A] curves and the Schild plots were found to have slopes greater than unity. The observed deviations from simple competitive antagonism appeared to be different from those reported earlier, and a possible explanation involving tissue-dependent noncompetitive factor(s) is discussed. PMID- 8243535 TI - Forskolin inhibition of K+ current in pregnant rat uterine smooth muscle cells. AB - Two kinds of outward K+ currents were examined in single smooth muscle cells from pregnant rat uterus, using whole-cell voltage clamp. The first and faster component was more sensitive to 4-aminopyridine (4-AP), whereas the second and slower (delayed) component was more sensitive to tetraethylammonium (TEA). A possible third K+ component (Ca activated K+ current) was not recorded as the pipette solution included EGTA. Forskolin inhibited the outward current in a concentration-dependent manner (50% inhibition occurred at about 30 microM); it affected the delayed component rather than the fast component. 8-Bromo-cAMP did not alter the outward current. In addition, inhibitors of protein kinase A and GDP-beta S and GTP-gamma S did not affect the forskolin-induced inhibition. These results indicate that forskolin inhibition of the delayed component of the outward current is independent of cAMP generation in the pregnant rat myometrial cells. Therefore, forskolin seems to directly inhibit specific K+ channels, as was reported for several other cell types. PMID- 8243536 TI - Structure-activity requirements of bombesin for gastrin-releasing peptide- and neuromedin B-preferring bombesin receptors in rat brain. AB - The pharmacological profile of [125I][Tyr4]bombesin binding to gastrin-releasing peptide- and neuromedin B-preferring sites has been investigated in rat cerebral cortex and olfactory bulb membranes, respectively. [125I][Tyr4]bombesin specific binding to cerebral cortex membranes was displayed biphasically by gastrin releasing peptide and [D-Phe6]bombesin-(6-13)-ethyl amide. In the presence of 10 mM neuromedin B, displacement curves for bombesin-related peptides were monophasic with gastrin releasing peptide displaying approximately 100-fold higher affinity than neuromedin B. In olfactory bulb membranes, [125I][Tyr4]bombesin binding was also displaced biphasically by gastrin releasing peptide, [D-Phe6]bombesin-(6-13)-ethyl amide and neuromedin B. In the presence of 10 microM [D-Phe6]bombesin-(6-13)-ethyl ester, displacement curves were monophasic with neuromedin B possessing approximately 10-fold higher affinity than gastrin-releasing peptide. Under these conditions, successive deletion of N terminal amino acids from bombesin-(1-14) was well tolerated at both sites, with little loss in affinity up to bombesin-(5-14). A 5- to 10-fold drop in affinity was observed at both sites with bombesin-(6-14), whilst the octapeptide acetyl bombesin-(7-14) displayed similar affinities to bombesin-(1-14). Bombesin-(8-14), -(9-14) and -(10-14) were essentially inactive (IC50 > 10 microM). C-terminal deletion of Met24 (bombesin-(1-13)) resulted in 100-fold loss of affinity at the gastrin-releasing peptide site and complete loss of affinity at the neuromedin B site. Fragments smaller than bombesin-(1-13) were virtually inactive at either site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243537 TI - L-deprenyl confers specific protection against MPTP-induced Parkinson's disease like movement disorder in the goldfish. AB - Administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to the goldfish causes a reversible, Parkinson's disease-like syndrome which includes loss of noradrenaline and dopamine from the brain, accumulation of the toxic metabolite 1-methyl-4-phenylpyridinium species (MPP+), and substantial reduction in movement. L-Deprenyl, a selective monoamine oxidase-B inhibitor, protects the goldfish from loss of movement, but clorgyline, a selective monoamine oxidase-A inhibitor, has no such protective action. L-Deprenyl and clorgyline primarily inhibit goldfish brain monoamine oxidase-B and monoamine oxidase-A, respectively. The mechanism by which MPTP causes reduced movement in goldfish is to cause an increase in resting time. Otherwise normal average velocity occurred during periods of movement. L-Deprenyl protection results in entirely 'normal' levels of resting time and average velocity during times of movement. Equivalent observations regarding l-deprenyl and clorgyline have been made in primate models of MPTP toxicity, and l-deprenyl is used for treatment of Parkinson's disease in humans. Therefore it is suggested that the evolutionarily equivalent subcortical circuitry and neural density of the goldfish brain may provide a useful model upon which to search for drugs relevant to human Parkinson's disease. PMID- 8243538 TI - Involvement of alpha-adrenoceptors in the endothelium-dependent depression of noradrenaline-induced contraction in rat aorta. AB - The involvement of endothelium-derived nitric oxide (NO) in the depressant action of the endothelium on noradrenaline-induced contractions and characterization of the receptor involved in the release of NO were studied using rat aorta. The noradrenaline-induced contraction was significantly potentiated by endothelium removal and in the presence of NG-nitro-L-arginine (L-NNA) or NG-monomethyl-L arginine (L-NMMA). The contraction induced by phenylephrine was also potentiated in the presence of L-NNA. Clonidine could induce contraction only in endothelium denuded preparations or in the presence of L-NNA. The potentiating action of L NNA on noradrenaline-induced contractions could also be observed in the presence of yohimbine or rauwolscine, although dose-response curves were shifted to the right. The depression of noradrenaline-induced contractions observed in the presence of the endothelium was increased by repeated stimulation. The depression was prevented by L-NNA and this effect was reversed by L-arginine. These results indicate the possibility that NO can be released through stimulation of alpha 1- and alpha 2-adrenoceptors on the endothelium and depresses noradrenaline-induced contractions of smooth muscle, although the contribution of the respective adrenoceptors remains to be investigated. The release of NO was increased when the stimulation was applied repeatedly. PMID- 8243539 TI - Inhibition of angiotensin-converting enzyme reduces urinary albumin excretion but not regional albumin clearance in experimental diabetes. AB - Albuminuria and increased regional albumin clearance are known to develop concomitantly in diabetes. The anti-proteinuric effect of angiotensin-converting enzyme inhibitor therapy is well established. We studied whether angiotensin converting enzyme inhibitor therapy with perindoprilat in comparison with conventional antihypertensive treatment could influence the development of increased regional albumin clearance as well as albuminuria in experimental diabetes. Rats with streptozotocin-induced diabetes were randomized into a saline group (n = 7), a perindoprilat (1 mg/kg per day) group (n = 8), and a hydralazine (3 mg/kg per day) group (n = 6); six rats served as non-diabetic controls. After 6-8 weeks, blood pressure was equally reduced in the perindoprilat- and hydralazine-treated groups (P < 0.01). Twenty-four-hour urinary protein and albumin excretion were increased in diabetic rats compared to control rats (P < 0.001). Hydralazine did not reduce 24 h protein or albumin excretion, whereas perindoprilat treatment reduced both (P < 0.001) to levels comparable to those of control rats. Regional albumin clearance, assessed in the eye, ileum, lung, skeletal muscle and skin, was clearly elevated in diabetic rats compared to control rats; however, neither drug therapy had an effect on albumin clearance. PMID- 8243540 TI - Blocking effect of serotonin on beta-adrenoceptor activity in follicle-enclosed Xenopus oocytes. AB - The effects of serotonin (5-HT) on a beta-adrenergic response were studied, using the voltage clamp technique, in Xenopus laevis oocytes surrounded by their follicular cells. noradrenaline induced marked hyperpolarization, with a specific increase in the permeability of the membrane toward K+. Application of 10 microM 5-HT had little effect on the resting membrane, but significantly depressed the response to 0.1 microM noradrenaline. The dose-response curve for noradrenaline showed a parallel shift to the right in the presence of 5-HT, suggesting that 5 HT competes with noradrenaline for common binding sites at the beta-adrenoceptor. PMID- 8243541 TI - Autoradiographic localization of [3H]nicardipine binding sites in the human renal artery. AB - In the present study the pharmacological profile and the anatomical distribution of dihydropyridine-type Ca2+ channels were analyzed in sections of the human renal artery by the use of combined radioligand binding and autoradiographic techniques with [3H]nicardipine as a ligand. The binding of [3H]nicardipine to sections of renal artery was time-, temperature- and concentration-dependent belonging, at least in the range of radioligand concentrations used, to a single class of high-affinity binding sites. The dissociation constant (KD) value was 0.3 nM and the maximum density of binding sites (Bmax) was 248 +/- 16 fmol/mg tissue. The pharmacological profile of [3H]nicardipine binding to sections of human renal artery was consistent with the labeling of dihydropyridine-type Ca2+ channels. In fact, dihydropyridine derivatives were the most powerful competitors of [3H]nicardipine binding, whereas phenylalkilamine, benzothiazepine or non selective channel modulators were weak or ineffective competitors. Light microscope autoradiography revealed the highest density of [3H]nicardipine binding sites in the tunica media of the renal artery, probably within smooth muscle cells. A smaller accumulation of the radioligand occurred in the tunica adventitia, whereas the tunica intima did not show specific binding. These results indicate that light microscope autoradiography techniques associated with radioligand binding may represent a useful tool for analyzing the localization of receptors or targets of drug action within the arterial wall. PMID- 8243542 TI - Glibenclamide, but not class III drugs, prevents ischaemic shortening of the refractory period in guinea-pig hearts. AB - The effective refractory period was measured in paced (4 Hz) perfused guinea-pig hearts in vitro. The effective refractory period was linearly correlated with temperature of the perfusing solution: as temperature was reduced the effective refractory period was increased. Reduction of the coronary flow rate to 10% of control resulted in a marked reduction in the effective refractory period. UK 66,914, dofetilide, ibutilide and phentolamine caused a prolongation in the effective refractory period, but during ischaemia the effective refractory period was reduced by the same degree as in vehicle-treated hearts. Glibenclamide had no effect on the effective refractory period prior to ischemia but it abolished the ischaemia-induced shortening. These results suggest that the opening of KATP channels may be responsible for the ischaemia-induced shortening of the effective refractory period in perfused guinea-pig hearts and that the class III effects of UK-66,914, dofetilide and ibutilide are attenuated during ischaemia. PMID- 8243543 TI - Hyperpolarization-activated inward current in embryonic chick cardiac myocytes: developmental changes and modulation by isoproterenol and carbachol. AB - Modulation of the hyperpolarization-activated inward current (If) in embryonic chick ventricular myocytes was examined using whole-cell voltage-clamp. Long (3 s) hyperpolarizing pulses were applied from a holding potential of -30 mV to steps of -40 to -120 mV. If was marked in 3-day-old cells, diminished at 10 days, and was almost completely gone at 17 days. If current density (at -120 mV) was 6.7 +/- 1.3 (3 days), -3.3 +/- 1.0 (10 days), and -2.0 +/- 0.5 pA/pF (17 days). If reduction paralleled the decrease in spontaneous activity. In 3-day cells, the threshold potential was -50 to -60 mV, and the reversal potential was -13.4 +/- 1.3 mV. The time course of activation was fitted by a single exponential and was temperature dependent: tau was 1.3 +/- 0.4 s at 20 degrees C and 0.7 +/- 0.4 s at 30 degrees C (at -120 mV). If amplitude was enhanced by 12.1 +/- 1.8% at 30 degrees C compared with 20 degrees C. Cs+ (3 mM) blocked If and had a negative chronotropic effect (rate decreased by 61%). Isoproterenol (1 microM) caused a positive chronotropic effect (17.1 +/- 2.9%) and increased If by 65.2 +/- 5.6%. Carbachol (0.1 microM) had a negative chronotropic effect (26.3 +/- 3.4%), and decreased If by 41.2 +/- 1.3%; it also reversed the enhancement produced by isoproterenol. Intracellular application of 100 microM GTP-gamma S decreased basal If by 35.2 +/- 5.0%, but potentiated the stimulant effect of isoproterenol (by 37.8 +/- 4.7%) and the inhibitory effect of carbachol (21.2 +/- 4.3%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243544 TI - Incomplete inhibition of endothelin-1 pressor effects by an endothelin ETA receptor antagonist. AB - Incomplete inhibition of endothelin-1-induced pressor effects by FR-139317, a novel, potent, ETA receptor antagonist, was observed in conscious, normotensive rats. Maximum inhibition by FR-139317 of the endothelin-1 pressor response (0.1, 0.3, 1.0 nmol/kg) was 49 +/- 7, 41 +/- 3, 62 +/- 5%, respectively. Two ETB selective receptor ligands induced pressor responses in conscious rats. A portion of the endothelin-1 pressor response may be mediated by ETB receptors, and ETB mediated vasoconstriction may contribute to incomplete inhibition of the pressor response to endothelin-1 by an ETA-selective receptor antagonist. PMID- 8243545 TI - Vasodilator action of cromakalim on human mesenteric artery. AB - Human mesenteric artery rings (1-3 mm diameter obtained from bowel resections), precontracted with KCl 80 mM, were relaxed by cromakalim (IC50 = 0.39 +/- 0.04 microM, maximum inhibition 69 +/- 2%). Cromakalim was more effective at inhibiting KCl 40 mM than KCl 80 mM. Cromakalim also inhibited contraction produced by noradrenaline, and was a more potent inhibitor of the second phase of the noradrenaline contraction than of the first phase. PMID- 8243546 TI - WAY 100,135 and (-)-tertatolol act as antagonists at both 5-HT1A autoreceptors and postsynaptic 5-HT1A receptors in vivo. AB - In binding studies, WAY 100,135 (N-tertiobutyl-3-[4-(2-methoxyphenyl) piperazinyl]-2-phenylpropana mide) and (-)-tertatolol showed affinities (Ki) of 29 nM and 10 nM, respectively, at 5-HT1A receptors. In vivo, they both dose dependently blocked the flat-body posture and corticosterone secretion provoked by an action of the 5-HT1A receptor agonist, S 14671 (1-[2-(2-thenoyl amino)ethyl]-4-[1-(7- methoxynaphtyl)]piperazine), at postsynaptic 5-HT1A receptors. Alone, they exerted little effect. The firing rate of dorsal raphe neurones, which bear inhibitory 5-HT1A autoreceptors, was reduced by S 14671 whereas it was not affected by WAY 100,135 and was increased by (-)-tertatolol. Both WAY 100,135 and (-)-tertatolol blocked the ability of S 14671 to inhibit raphe firing. In conclusion, these data demonstrate that WAY 100,135 and (-) tertatolol behave as antagonists at both 5-HT1A autoreceptors and postsynaptic 5 HT1A receptors in vivo. PMID- 8243547 TI - Dose-response differences in the ability of ramipril to improve retention in diabetic mice. AB - Ramipril blocks the conversion of angiotensin I to II. The literature indicates that diabetes is often associated with mild impairment of learning and memory. The study reports the effects of ramipril on memory retention in diabetic and non diabetic mice. Mice were made diabetic by an injection of streptozocin. After overt signs of diabetes were present, diabetic or vehicle-treated mice were partially trained on a footshock active avoidance task. Immediately after training, ramipril (0.5-1.5 mg/kg s.c.) was administered and retention was tested by continuing training one week later until mice avoided footshock on five out of six trails. The results indicate that ramipril enhanced retention of both diabetic and control mice but it required about 5 times as much ramipril in diabetic as control mice to achieve the same effect on retention. Increased sensitivity to angiotensin II may play a role in cognitive impairment in diabetes. PMID- 8243548 TI - Relaxation by dexamethasone of isolated guinea-pig airways precontracted with endothelin-1. AB - Dexamethasone (1-100 microM) produced concentration-dependent relaxations of isolated guinea-pig tracheal and bronchial strips precontracted with endothelin-1 (100 nM), which were not affected by the removal of the epithelium. Dexamethasone neither induced relaxation of tissues precontracted with carbachol (0.1 microM) or KCl (25 mM) nor inhibition of endothelin-1 (1-100 nM)-induced peak contractions after 10-min preincubation of the tissues with dexamethasone. These findings indicate that acute corticosteroid treatment can reverse the bronchoconstrictor action of endothelin-1. PMID- 8243549 TI - Modification of the N-methyl-D-aspartate response by antidepressant sigma receptor ligands. AB - Sertraline, a selective serotonin reuptake inhibitor, and clorgyline, a monoamine oxidase inhibitor, both of which have high affinity for sigma receptors, were assessed in an electrophysiological model. In keeping with previous data obtained with other sigma receptor ligands, low doses of sertraline and of clorgyline potentiated selectively with a bell-shaped dose-response curve the effect of N methyl-D-aspartate (NMDA) on pyramidal neurons in the CA3 region of the rat dorsal hippocampus. This potentiation was reversed by the sigma receptor ligands haloperidol and BMY-14802. The selective serotonin reuptake inhibitor paroxetine and the monoamine oxidase inhibitor tranylcypromine, both devoid of affinity for sigma receptors, had no effects on the NMDA response. These data suggest that the effects of sertraline and clorgyline on the NMDA response are due to their affinity for sigma receptors. PMID- 8243550 TI - Co-administration of (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid and arachidonic acid potentiates synaptic transmission in rat hippocampal slices. AB - Perfusion of the 1S,3R isomer of trans-aminocyclopentane-1,3-dicarboxylic acid (t ACPD, 50 microM), or arachidonic acid (10 microM), for 5 min produced only depression of the field excitatory postsynaptic potential recorded in the CA1 region of rat hippocampal slices from which the CA3 region had been removed. However, perfusion of t-ACPD and arachidonic acid in combination induced a rapid potentiation of the response which in 4/6 slices was maintained for at least 90 min. PMID- 8243551 TI - Analysis of the mechanisms underlying the biphasic responses to bradykinin in circular muscle from guinea pig ileum. AB - Experiments were designed to further characterize the receptor mediating the biphasic response to bradykinin in circular muscle from guinea pig ileum in vitro by the use of HOE 140, a potent and specific bradykinin antagonist. D-Arg [Hyp3,Thi5,D-Tic7,Oic8]bradykinin (HOE 140, 0.1-1000 nM) caused a graded inhibition of bradykinin (10 nM)-induced contraction and relaxation responses in circular muscle from guinea pig ileum, with IC50s of 4 and 10 nM respectively. However, the potency of HOE 140 to antagonize the bradykinin (300 nM)-induced contraction and relaxation was decreased about 6-fold (IC50 22 nM) and 57-fold (IC50 570 nM). HOE 140 (3-100 nM) caused parallel and concentration-dependent rightward displacements of bradykinin (0.1-3000 nM)-induced biphasic concentration-response curves in circular muscle from guinea pig ileum. Schild regression plots yielded straight lines with slopes not significantly different from unity and pKb values of 9.0 and 8.7 against bradykinin-induced contraction and relaxation, respectively. Similar pKb values (8.7) were obtained for HOE 140 against bradykinin-mediated contraction in the longitudinal muscle of the guinea pig ileum. The action of HOE 140 was selective for bradykinin, since response to other agonists were not affected. It is concluded that HOE 140 does not discriminate the receptors mediating the biphasic responses to bradykinin in circular muscle from guinea pig ileum, as it showed a similar selective, competitive and reversible antagonism against both components of the bradykinin response in this preparation. PMID- 8243552 TI - Comparative effects of the two endothelin ETA receptor antagonists, BQ-123 and FR139317, on endothelin-1-induced contraction in guinea-pig iliac artery. AB - The effects of two recently introduced endothelin ETA receptor antagonists, BQ 123 and FR139317, were investigated and compared in guinea-pig isolated iliac artery. Endothelins and sarafotoxins induced contraction of guinea-pig iliac artery with a pharmacological profile characteristic of the ETA receptor. The rank order of agonist potency was (mean EC50 values, nM): endothelin-1 (11.7) > or = endothelin-2 (14.9) > or = vasoactive intestinal contractor (19.5) > sarafotoxin S6b (49.8) > or = [Ala3,11]endothelin-1 (55.0) > sarafotoxin S6a (> 100) > endothelin-3 (> or = 1000). The C-terminal hexapeptide, endothelin-(16 21), sarafotoxin S6c and sarafotoxin S6d were neither agonists nor antagonists at concentrations up to 10, 3 and 1 microM, respectively. Both FR139317 (1-10 microM) and BQ-123 (0.1-1 microM) surmountably antagonized the effects of endothelin-1. Schild analysis suggested competitive antagonism for FR139317 (Schild slope 1.32 +/- 0.21, pA2 5.82 +/- 0.16, n = 5), but not for BQ-123 (Schild slope 0.28 +/- 0.08, n = 5), which was however more potent (apparent pKB 6.6-7.2) than FR139317. The potency of FR139317 was particularly low with respect to the reported affinity for ETA receptors, suggesting heterogeneity among ETA receptors. Thus, the endothelin receptor present in guinea-pig iliac artery has the following features: (1) rank order of agonist potencies of the ETA type; (2) low potency of FR139317 and (3) non-competitive antagonism by BQ-123. PMID- 8243553 TI - Vasorelaxant action of Ki1769, a new pyridinecarboximidamide, in isolated porcine coronary artery. AB - The characteristics of KRN2391 (N-cyano-N'-(2-nitroxyethyl)-3 pyridinecarboximidamide monomethansulfonate) and its phenethyl and 2-hydroxyethyl derivatives (Ki1769 and Ki3315) were studied in isolated porcine coronary arteries. KRN2391, Ki1769 and Ki3315 produced concentration-dependent relaxation of coronary arteries contracted by 25 mM KCl and the order of relaxant potency was KRN2391 > Ki1769 > Ki3315. At the maximum effect, KRN2391 produced nearly complete relaxation but Ki1769 produced about 66% relaxation. The maximum effect of Ki3315 could not be obtained because of its solubility. The relaxation induced by KRN2391 was antagonized by glibenclamide and methylene blue but relaxations caused by Ki1769 and Ki3315 were antagonized by glibenclamide alone. The antagonistic effect of glibenclamide on Ki1769- and Ki3315-induced relaxations was more potent than that on KRN2391-induced relaxation. KRN2391 induced relaxation of coronary arteries contracted by 40 mM KCl in a concentration dependent manner but the effect of KRN2391 was smaller against 40 mM KCl-induced contractions than against 25 mM KCl-induced contractions. Ki1769 had almost no effect on coronary arteries contracted by 40 mM KCl. These results suggest that pyridinecarboximidamide derivatives which do not possess a nitroxyl group have vasodilating ability based on a K+ channel opening action. PMID- 8243554 TI - Hypouricemic effect of the novel xanthine oxidase inhibitor, TEI-6720, in rodents. AB - We investigated the xanthine oxidase/xanthine dehydrogenase inhibitory activity and hypouricemic effect of a newly synthesized xanthine oxidase/xanthine dehydrogenase inhibitor, TEI-6720, 2-(3-cyano-4-isobutoxyphenyl)-4-methyl-5 thiazole-carboxylic acid, and compared its effects with those of allopurinol in rodents. TEI-6720 was found to inhibit bovine milk xanthine oxidase, and mouse liver and rat liver xanthine oxidase/xanthine dehydrogenase with IC50 values of 1.4, 1.8 and 2.2 nM, respectively. On bovine milk xanthine oxidase, TEI-6720 exhibited mixed-type inhibition and the Ki value was 0.7 nM. TEI-6720 displayed prolonged urate-lowering activity in normal mice and rats. We evaluated the hypouricemic effect of TEI-6720 on hyperuricemia induced by the uricase inhibitor, potassium oxonate (250 mg/kg s.c., 1 h before the test drugs), and measured the total molarity of both serum allantoin and urate in rats. Oral TEI 6720 and allopurinol had a hypouricemic effect 2 h after their administration to oxonate-pretreated rats with ED50 values of 1.5 and 5.0 mg/kg, respectively. Both compounds also reduced the combined molarity of uric acid and allantoin in rats. The ED50 values of TEI-6720 and allopurinol were 2.1 and 6.9 mg/kg p.o., respectively. These results suggest that TEI-6720 may be useful for the treatment of hyperuricemia. PMID- 8243555 TI - Localization and characteristics of atrial natriuretic peptide receptors in prenatal and postnatal rat brain. AB - We studied the expression of atrial natriuretic peptide (ANP) receptor subtypes during development in the rat forebrain, using quantitative autoradiography. Highest ANP binding was observed in the cortical neuroepithelium at embryonic day 17. Lower ANP binding was found in cingulate and frontal cortices at postnatal day 10, but none was detectable at 8 weeks of age. In the neuroepithelium of the embryonic rat, binding was displaced with a potency of rat ANP-(1-28) (rANP) > porcine type-C natriuretic peptide (pCNP-22) = rat ANP fragment C-ANP-(4-23) (rC ANP-(4-23)) = rat brain natriuretic peptide (rBNP-32), different from that of any of the well-characterized (ANPA, ANPB, and ANPC) natriuretic peptide receptors. The present results support the hypothesis of a role for ANP during brain maturation and indicate that the ANP receptors highly expressed in the embryonic neuroepithelium may belong to a new ANP receptor subtype not yet characterized. PMID- 8243556 TI - Cardioprotection by ramiprilat in isolated rabbit hearts. AB - The anti-ischemic properties of the ACE inhibitor ramiprilat (ram) were investigated in electrically driven Langendorff hearts from rabbits whose endogenous angiotensin-I content has been previously shown to be very low (constant pressure: 70 cm H2O, Tyrode solution, Ca2+ 1.8 mmol/l). Cumulative concentration-response curves showed that the reduction in global coronary flow (CF) by exogenous angiotensin-I was concentration dependently inhibited by ram (P < 0.05). Myocardial ischemia (MI) was induced by occlusion of a left coronary artery branch and MI was quantified by NADH surface fluorescence photography. MI was significantly enlarged (+23%) (P < 0.05) by exogenous angiotensin-I (6 x 10( 9) mol/l). Addition of ram (10(-8) mol/l) to the perfusion buffer simultaneously with angiotensin-I, completely prevented the reduction of CF by angiotensin-I (P > 0.05) and significantly diminished MI even below control values (-25%) (P < 0.05). In the absence of exogenous angiotensin-I, ram alone (10(-8) mol/l) did not significantly enhance CF (P > 0.05), supporting findings demonstrating a very low endogenous angiotensin-I content in isolated rabbit hearts. However, ram alone (10(-8) mol/l) significantly diminished MI (-24%) (P < 0.05). We conclude that ram does possess direct cardioprotective properties that are independent of the inhibition of angiotensin-II generation but that may be related to potentiation of the effects of bradykinin. PMID- 8243557 TI - Excitation-contraction uncoupling in the rat diaphragm by 2,3-butanedione monoxime. AB - 2,3-Butanedione monoxime (BDM), 0.5-12.5 mM, inhibited the twitches rapidly and reversibly in the rat diaphragm. A more extensive inhibition at direct than at indirect stimulation, a simultaneous effect on the electromyogram, and an attenuation of the inhibitory effect in preparations made hyperexcitable with Ca(2+)-free solution, myotonia or veratridine, indicated an interference with the gating mechanism of the sarcolemma. The excitation-contraction uncoupling with BDM was additive to the similar effect of dantrolene. A synergistic effect was not found, and neither agent alone nor both in combination caused a complete excitation-contraction uncoupling. Several differences between the effects of BDM and dantrolene were disclosed: Lowering the temperature increased and decreased the effect of BDM and dantrolene, respectively. Tetanic stimulation and high Ca2+ or high K+ antagonized the effect of dantrolene and did not affect the BDM induced inhibition. Ca(2+)-free solution potentiated the effect of dantrolene and antagonized the effect of BDM. Caffeine and KCl contractures were inhibited by BDM, whereas the propranolol contracture due to Na,K-ATPase inhibition, which is completely inhibited by dantrolene, was unaffected by BDM. PMID- 8243558 TI - Effect of acute and short-term administration of cholinomimetic drugs on corticosterone secretion in the rat. AB - Centrally acting cholinomimetic drugs have been proposed for the therapy of cognitive disorders in aged subjects. Among the possible adverse side effects of this class of compounds, of great relevance is the stimulatory action on the adrenocortical axis, in view of the toxicity of glucocorticoids for hippocampal neurons and the immune system. The aim of the present study was to evaluate in conscious male rats the effect of acute and short-term administration of three novel cholinomimetic drugs on the release of corticosterone. The potent agonist of muscarinic receptors RU 35963 strikingly increased corticosterone levels after acute but not after short-term (6 days) administration. Similar results were obtained after administration of the reversible inhibitor of cholinesterase, eptastigmine. In contrast to RU 35963 and eptastigmine, acute administration of a choline precursor, L-alpha-glycerylphosphorylcholine, only slightly affected plasma corticosterone concentrations after both acute and short-term administration. It is concluded that activation of adrenocortical function by cholinomimetic drugs is a short-lasting event which does not represent an important side effect of these compounds when given on a long-term basis. PMID- 8243559 TI - Differential effects of hydroxocobalamin on relaxations induced by nitrosothiols in rat aorta and anococcygeus muscle. AB - In aortic rings, hydroxocobalamin (30 microM) reduced the relaxant actions of S nitrosocysteine (0.1-3 microM), S-nitrosoglutathione (0.1-3 microM) and S-nitroso N-acetylpenicillamine (SNAP, 0.01-3 microM), but did not affect the relaxant action of S-nitroso-coenzyme A (0.1-3 microM). In anococcygeus muscles, hydroxocobalamin (30 microM) had little effect on relaxations produced by nitrosocysteine (0.1-3 microM) and SNAP (0.01-1 microM), and enhanced those produced by nitrosoglutathione (0.1-3 microM) and nitroso-coenzyme A (0.1-3 microM). Since hydroxocobalamin is thought to act like haemoglobin by sequestering NO, some of the effects of hydroxocobalamin were compared with those of haemoglobin. Haemoglobin (10 microM) inhibited relaxations of aortic rings produced by nitrosocysteine and nitrosoglutathione and relaxations of anococcygeus muscles produced by nitrosocysteine, nitrosoglutathione and SNAP. Thus the effects of hydroxocobalamin on nitrosothiol-induced relaxations differ between the rat aorta and anococcygeus muscle, and depend on the exact nature of the nitrosothiol; however, the effects of haemoglobin did not differ qualitatively between the two tissues. Since hydroxocobalamin reduced relaxations of rat anococcygeus muscles elicited by NO, but not those elicited by nitrergic nerve stimulation or nitrosothiols, the nitrergic transmitter more closely resembles a nitrosothiol than free NO. Of those tested, the best correspondence was with nitrosocysteine; however, there were some differences between it and the transmitter. PMID- 8243560 TI - Acute noise stress reduces [3H]5-hydroxytryptamine uptake in rat brain synaptosomes: protective effects of buspirone and tianeptine. AB - Acute noise stress decreased [3H]5-hydroxytryptamine ([3H]5-HT) uptake in synaptosomes from rat hypothalamus, hippocampus and cerebral cortex. The decrease was due to the maximum rate of [3H]5-HT uptake, which peaked 30 min after stress and partly returned to resting values within 4 h, with no changes in affinity (Km values). No changes in [3H]paroxetine binding and basal [3H]5-HT release were found in stressed rats. Tianeptine, given at the dose of 10 mg/kg 1 h before stress, counteracted the noise-induced decrease of 5-HT uptake, since it increased [3H]5-HT uptake in both resting and stressed animals, but did not prevent the rise in plasma corticosterone of stressed rats. Buspirone pretreatment had no effect on [3H]5-HT uptake in resting rats but prevented the noise-induced decrease in [3H]-HT uptake. Diazepam did not modify either the basal or the noise-induced reduction in [3H]5-HT uptake. The evidence that treatments reducing extrasynaptic 5-HT, by increasing its reuptake (tianeptine) or reducing its release (buspirone) in innervated regions are able to modify the stress-induced decrease in 5-HT uptake, further confirms the importance of serotonin in the mechanisms mediating neurochemical responses to stress. PMID- 8243561 TI - Inhibitory effects of ibogaine on cocaine self-administration in rats. AB - In order to determine the potential anti-addictive properties of ibogaine, we used the cocaine self-administration model in rats. The results indicate that a single injection of ibogaine (40 mg/kg i.p.) produced a significant decrease of cocaine intake, which remained unaltered for more than 48 h. Since the half-life time of ibogaine is short, this might suggest the involvement of one or several active metabolites of ibogaine in cocaine intake. Repetitive administration of ibogaine on three consecutive days also induced a pronounced decrease of cocaine intake. However, a more prominent inhibitory effect on cocaine intake was observed in animals treated repeatedly with ibogaine (40 mg/kg i.p.), once each week for 3 consecutive weeks. These results indicate that ibogaine or its metabolite(s) is a long-lasting interruptor of cocaine dependence, which supports similar observations from uncontrolled clinical studies. PMID- 8243562 TI - Hemodynamic effects of a novel hypotensive peptide, human adrenomedullin, in rats. AB - The hemodynamic effects of human adrenomedullin were investigated in anesthetized Wistar rats. Intravenous administration of adrenomedullin (1.0 nmol/kg) caused a rapid and marked reduction in mean blood pressure associated with a decrease in total peripheral resistance. This reduction in mean blood pressure was closely correlated with the decrease in total peripheral resistance. These findings indicate that human adrenomedullin is a potent vasodilator and may have some role in the regulation of blood pressure. PMID- 8243563 TI - Repeated exposure to cocaine produces long-lasting deficits in the serotonergic stimulation of prolactin and renin, but not adrenocorticotropin secretion. AB - To determine whether cocaine-induced deficits in serotonergic function are long lasting, the neuroendocrine responses to the serotonin (5-hydroxytryptamine, 5 HT) releaser, p-chloroamphetamine were evaluated 1-8 weeks subsequent to 7 days of cocaine exposure (15 mg/kg b.i.d.). In cocaine-pretreated rats, the p chloroamphetamine-induced elevations of prolactin and renin secretion were significantly reduced for 8 and 4 weeks, respectively. In contrast, the p chloroamphetamine-induced elevation of adrenocorticotropic hormone (ACTH) secretion was at control values 1 week after cocaine exposure. The data suggest that some cocaine-induced deficits in serotonergic function are long-lasting. PMID- 8243564 TI - Continuous infusion of interleukin-6 in sublethally irradiated mice accelerates platelet reconstitution and the recovery of myeloid but not of megakaryocytic progenitor cells in bone marrow. AB - Interleukin-6 (IL-6) is a pleiotropic cytokine that enhances the maturation of megakaryocytes. In mice, in vivo treatment with IL-6 results in elevated platelet counts both in untreated animals and after myelosuppressive therapy. In this study, we assessed the effect of continuous infusion of IL-6 in sublethally irradiated (7 Gy) mice on peripheral blood cell counts and progenitor cells in bone marrow and spleen. Female Swiss mice were treated by continuous infusion with 1 or 10 micrograms IL-6 per day for 7 or 14 days. Continuous infusion of IL 6 for 7 days resulted in elevated levels of circulating IL-6 (mean: 1872 pg/mL vs. 100 pg/mL for phosphate-buffered saline [PBS]-treated controls) and in an accelerated reconstitution of platelets starting at day 12 after irradiation. In IL-6-treated animals, the 50% pretreatment platelet count was reached on day 15 vs. day 21 for irradiated controls receiving no IL-6. Treatment with IL-6 for 14 days resulted in a further increase in platelet counts, exceeding the pretreatment counts. The number of colony-forming units-megakaryocyte (CFU-Mk) was significantly elevated from day 6 to 18 in the spleen but not in bone marrow. To assess the contribution of extramedullary megakaryocytopoiesis in the spleen to IL-6-induced platelet recovery, IL-6 was also administered to splenectomized mice. The stimulatory effect of IL-6 on platelet recovery was preserved in these animals, indicating that megakaryocytopoiesis in the spleen did not contribute to the accelerated recovery of platelets. The neutrophil counts were elevated during IL-6 treatment and became similar to controls after cessation of therapy, whereas the numbers of colony-forming units-granulocyte/macrophage (CFU-GM) in the bone marrow were elevated from day 9 to 24 in all animals treated with 10 micrograms IL-6 per day. In conclusion, continuous infusion of IL-6 stimulates platelet recovery after irradiation without increasing the number of CFU-Mk and conversely stimulates the proliferation of myeloid progenitor cells without an effect on neutrophil reconstitution. PMID- 8243565 TI - Apoptosis in tumor necrosis factor-alpha-dependent, monocyte-mediated leukemic cell death: a functional, morphologic, and flow-cytometric analysis. AB - Little is known about the precise ways in which monocytes and macrophages recognize tumor cells and how they exert their cytolytic and/or cytostatic effects. By a functional, morphologic, and flow-cytometric approach, we have studied monocyte/macrophage- and cytokine-mediated cytotoxicity against U937 cells, a human histiocytic lymphoma cell line. A rapid decrease in cell viability of U937 cells (MTT assay) could be observed at an effector-to-target cell (E:T) ratio of 10 in the presence of interferon (IFN)-gamma-activated monocytes. Light and electron microscopic examination showed the characteristic features of apoptosis of U937 cells after incubation with either monocytes or tumor necrosis factor (TNF)-alpha. TNF-alpha-induced apoptosis (10(4) U/mL) as measured by multiparameter flow cytometry (propidium iodide [PI]) paralleled the functional decrease in cell viability (MTT assay) of 20 +/- 3% after 24 hours up to a maximum of 50 +/- 4% after 48 hours. Apoptosis could be confirmed by the detection of DNA degradation into multiples of 200-bp subunits by agarose gel electrophoresis. After prolonged incubation times, monocyte-mediated leukemic cell death could be quantified as apoptosis by flow cytometry, whereas no decrease in net cell viability of tumor cells relative to the initial cell number could be observed by MTT spectrophotometry. In conclusion, our data provide evidence that apoptosis is the major mode of TNF-alpha-dependent monocyte mediated cytotoxicity against U937 cells. Furthermore, multiparameter flow cytometric analysis offers a sensitive method to quantify cytokine- and cell induced apoptosis in leukemia. PMID- 8243566 TI - Effect of combined treatment with interleukin-3 and interleukin-6 on 4 hydroperoxycyclo-phosphamide-induced programmed cell death or apoptosis in human myeloid leukemia cells. AB - In autologous bone marrow transplantation in patients with acute myeloid leukemia (AML), 4-hydroperoxycyclophosphamide (4-HC) is a commonly used ex vivo purging agent for leukemic blasts. In the present report, we demonstrate that exposure to high concentrations of 4-HC for 1 hour, as used in ex vivo bone marrow purging, produces internucleosomal DNA fragmentation characteristic of apoptosis, or programmed cell death (PCD), in human myeloid leukemia HL60 cells. Lower concentrations of 4-HC (10, 20, or 50 microM/L) failed to cause this effect, while higher concentrations (> or = 200 microM/L) produced random DNA fragmentation. 4-HC-mediated internucleosomal DNA fragmentation was associated with a marked induction in c-jun and significant reductions in bcl-2 and c-myc oncogene expressions. A combined treatment with interleukin-3 (IL-3) plus IL-6 for 18 hours before an additional, 1-hour concurrent treatment with 4-HC (100 microM/L) significantly increased 4-HC-induced DNA fragmentation as well as colony growth inhibition of HL60 cells. The effects of cotreatment with IL-3 plus IL-6 were also associated with a further, modest decrease in bcl-2 and c-myc and augmentation of c-jun expression. These findings highlight an alternative mechanism of 4-HC-induced leukemic cell death that can be potentially enhanced by cotreatment with IL-3 plus IL-6. PMID- 8243567 TI - A sensitive new bioassay for erythroid colony-stimulating factor. AB - Erythroid colony-stimulating factor (E-CSF) is a B cell-derived membrane protein that specifically affects the growth and development of human and murine committed erythroid progenitors. We report the development of a sensitive new bioassay for E-CSF, based on the ability of the growth factor to stimulate 3H thymidine incorporation into cloned Rauscher murine erythroleukemia cells. The assay has among its advantages the ability to measure growth factor activity on a purified target cell population in the absence of endogenous growth factor producing accessory cells. In addition, this assay measures E-CSF's proliferative effect on erythroid progenitors in the absence of erythropoietin (Epo) after 72 to 96 hours. In contrast, the standard bone marrow fibrin clot assay traditionally used to measure E-CSF requires the addition of Epo to promote the development of hemoglobinized erythroid colonies that are quantified after 7 days (for murine cells) to 12 days (for human cells). With the use of this new Rauscher cell bioassay, we have identified an E-CSF-producing human cell line and, further, have measured E-CSF activity derived from nonhuman splenic B lymphocytes. PMID- 8243568 TI - In vitro sensitivity of human hematopoietic progenitor cells to 4 hydroperoxycyclophosphamide. AB - Protracted engraftment and prolonged thrombocytopenia remain problems when marrow purged with 4-hydroperoxy-cyclophosphamide (4-HC) is employed for autotransplants. Toxic effects of 4-HC on early progenitor and stem cells might play an important causative role. Surprisingly, few investigations have examined the effects of 4-HC on progenitor cells other than colony-forming units granulocyte/macrophage (CFU-GM) and burst-forming units-erythroid (BFU-E), and only one study could be found where 4-HC exposure was carried out on cells purified beyond the buffy coat stage. Since the cellular milieu, and in particular the red blood cell count, can ameliorate 4-HC toxicity, the suppressive effect of this agent on marrow stem cells might be underestimated. We therefore investigated the relative 4-HC sensitivity of different human bone marrow progenitor cells in vitro using partially purified adherent cell- and T lymphocyte-depleted bone marrow mononuclear cells (A-T-MNC). Cells were exposed to increasing doses (10 to 200 micrograms/mL) of 4-HC using a standard purging protocol established for bone marrow buffy coat cells. Effects on mixed (CFU Mix), erythroid (CFU-E and BFU-E), granulocyte-macrophage (CFU-GM), and stromal cell (CFU-F) progenitors were determined. In addition, we examined the 4-HC sensitivity of megakaryocyte progenitors (CFU-Meg) since, to our knowledge, this has not been reported before. As expected, increasing doses of 4-HC led to progressive inhibition of hematopoietic colony formation. CFU-F, CFU-Mix, and CFU Meg appeared most resistant to 4-HC exposure, while CFU-E, BFU-E, and CFU-GM appeared most sensitive. At doses over 100 micrograms/mL, the usual concentration recommended for purging of buffy coat cells, hematopoietic colony formation was completely inhibited. These data suggest that if more highly purified marrow cells are employed for purging, lower 4-HC doses may need to be used. They also suggest that the thrombocytopenia that frequently accompanies 4-HC purging is not likely due to loss of CFU-Meg. PMID- 8243569 TI - Invalidity from nonparallelism in a radioimmunoassay for erythropoietin accounted for by human serum antibodies to rabbit IgG. AB - An immunologic crossreactant of erythropoietin seemed to develop and persist in serum samples from a patient during treatment and remission of idiopathic aplastic anemia. It had a steeper slope to radioimmunoassay log-dose response lines and a larger molecular size than erythropoietin. On fractionation of serum, the apparent crossreactant was bound by staphylococcal Protein A at pH 7.5 and recovered by elution from it at pH 3.0. Adsorption of serum from the patient, and from one of two similarly affected children, with rabbit IgG linked to agarose appeared to remove completely the apparent crossreactant. These treated sera gave radioimmunoassay log-dose response lines essentially parallel to that given by the International Reference Preparation (IRP) for erythropoietin and estimates of immunoreactive erythropoietin appropriate to the normal hemoglobin concentrations. The apparent crossreactant of erythropoietin is thus accounted for by heterophilic antibodies to rabbit IgG. These developed in the patient following treatment with rabbit antilymphocyte globulin but seem to have arisen spontaneously in the children. Thus iatrogenic and idiopathic antibodies to rabbit IgG interfered in a radioimmunoassay for erythropoietin in serum through their ability to react with the radioimmunoassay anti-erythropoietin antiserum raised in rabbits. PMID- 8243570 TI - Ribozyme-mediated cleavage of the bcr/abl transcript expressed in chronic myeloid leukemia. AB - The bcr/abl transcript is specifically expressed in the hematopoietic cells of patients with chronic myeloid leukemia (CML). The Haseloff and Gerlach model was used to design a ribozyme targeted to this transcript. When tested against a synthetic substrate covering the translocation sequence, the ribozyme exhibited site specificity and an absolute requirement for divalent metal ions. Cleavage of the normal bcr was also noted but at a reduced efficiency compared to that exhibited for the bcr/abl substrate. Importantly, cleavage of the full-length bcr/abl mRNA was achieved at physiologic temperature, demonstrating effective ribozyme-mediated cleavage. PMID- 8243571 TI - Detection and direct sequence identification of BCR-ABL mRNA in Ph+ chronic myeloid leukemia. AB - The reverse transcriptase-polymerase chain reaction (RT-PCR) for BCR-ABL mRNA is increasingly used to diagnose and monitor patients with Ph+ chronic myeloid leukemia (CML). We investigated an alternative approach to detect BCR-ABL mRNA in CML in order to overcome some of the potential drawbacks of RT-PCR. Nucleic acid sequence based amplification (NASBA) is a homogeneous, isothermal, in vitro process that provides the direct amplification of RNA. Peripheral blood from seven patients with Ph+ CML and Ph+ EM-2 cells were investigated by NASBA and RT PCR. A nested set of four primers flanking the BCR-ABL junction was used in two serial NASBA reactions performed for 2 hours. The two methods were fully concordant for detection of transcripts with bcr3-abl2 and bcr2-abl2 junctions. Ethidium bromide fluorescence with NASBA indicated in repeated experiments that similar quantities of total RNA from patient material contained different amounts of BCR-ABL mRNA. The data suggest that direct amplification of RNA is suitable for identifying and monitoring patients with Ph+ CML and may provide a means to quantify BCR-ABL mRNA levels. PMID- 8243572 TI - Reticular facilitation of cat visual cortical responses is mediated by nicotinic and muscarinic cholinergic mechanisms. AB - Stimulation of the mesencephalic reticular formation facilitates responses in the visual cortex elicited from the optic radiation. Using intravenous administration of cholinergic antagonists we investigated in adult cats and two kittens whether this effect is mediated by cholinergic mechanisms. When administered alone the muscarinic antagonists atropine and scopolamine and the nicotinic antagonist mecamylamine failed to block reticular facilitation and sometimes even enhanced the effects of reticular stimulation. However, when administered in combination muscarinic and nicotinic antagonists eliminated or significantly reduced the facilitation. This was even true when the two antagonists were administered with a time lag of several hours. These results support the notion that reticular facilitation of cortical responses is mediated by cholinergic mechanisms and suggest that this effect is mediated either by a receptor with a mixed pharmacological property or by two independent pathways acting via nicotinic and muscarinic receptors. This hypothesis is discussed in the context of recent evidence on cholinergic transmission and earlier data on the pharmacology of reticular arousal. PMID- 8243573 TI - The effect of gravity on the resting position of the cat's eye. AB - We measured rotation (horizontal, vertical and torisonal) and translation (horizontal and vertical) of the paralysed cat's eye in response to 45 degrees steps of orientation presented in a pseudorandom order around the roll and pitch axes (with respect to the horizontal canals). During changes of position of the animal in the roll plane, the eyes rotated towards the lowest part of the orbit (left with left ear down; top when the cat was upside down, etc.) by an average of 0.55 degree. Changing orientation in the pitch plane evoked vertical rotations of +/- 1.42 degrees (upwards eye movement during forwards head pitch) and torsional rotations of +/- 1.3 degrees. All these rotations taken together suggest that the centre of mass is in front of, below and temporal to the centre of rotation. The eyes translated temporally (thus separating by 0.72 mm) during forward pitching and there was a small vertical displacement (0.23 mm) when the animal was upside down. These findings are discussed with respect to a possible role of the extraocular proprioception system. PMID- 8243574 TI - An electrophysiological study of descending projections to the lumbar spinal cord in adult male rats. AB - Recent anatomical evidence suggests that descending projections from the lateral vestibular nucleus (LVe) and gigantocellular reticular nucleus (Gi) innervate areas of the lumbar spinal cord near the spinal nucleus of the bulbocavernosus (SNB). To confirm this finding electrophysiologically, we recorded and mapped averaged field potentials within the lumbar spinal cord of male rats in response to electrical stimulation of the LVe or Gi and compared these with the location of averaged field potentials evoked at the same levels by stimulation of SNB axons in the bulbocavernosus (BC) nerve. Stimulation of the LVe or the Gi produced negative field potentials that were largest at sites 200-450 microns dorsolateral to SNB somata. In an attempt to verify that this region innervates SNB motoneurons, the BC motor nerve volley was recorded in response to microstimulation at various depths within the spinal cord. Stimulation of sites dorsolateral and lateral to the SNB somata elicited volleys in the BC nerve that had two components. The onset latency of the earlier component was similar to the antidromic latency of SNB motoneurons to BC nerve stimulation, and the threshold for eliciting this component was lowest at sites in the electrode track near SNB somata. Thus, the earlier component may be evoked by direct stimulation of the SNB motoneurons. The threshold for evoking the later component was lowest at the sites 230-380 microns dorsolateral to SNB somata, suggesting that this component involves activation of other neurons. These results indicate that the LVe and Gi may modulate the activity of SNB motoneurons through interneurons located in a region several hundred microns away from SNB somata. PMID- 8243575 TI - The distal hindlimb musculature of the cat: interanimal variability of locomotor activity and cutaneous reflexes. AB - During stereotyped behaviors such as locomotion, patterns of muscle recruitment are usually quite consistent from animal to animal, even in the face of many surgical and pharmacological reductions. However, as studies of musculoskeletal structure, neuromuscular architecture, and sensorimotor circuitry become more detailed, it is important to ask whether there is some level of organization at which individual differences begin to dominate. This study concentrated on the small muscles of the foot and ankle, using standardized methods that consistently record stereotypical electromyographic activity from prime mover muscles and that permit well-calibrated stimulation of cutaneous nerves to elicit reflexes during treadmill locomotion. Some muscles (particularly the main ankle extensors, triceps surae, and plantaris) had stereotyped activity during both unperturbed locomotion and reflex responses. Others had stereotyped activity during locomotion but variable reflex patterns among animals (tibialis anterior, extensor digitorum longus, flexor hallucis longus, and peroneus brevis). Still others had variable locomotor activity but reflexes that were consistent (flexor digitorum longus) or variable for only peroneal nerve stimulation (peroneus longus), only plantar nerve stimulation (peroneus tertius), or the two (flexor digitorum brevis). Among muscles with interanimal variability, there seemed to be no particular correlation between locomotor and reflexive recruitment in a given animal. This functional heterogeneity is discussed in terms of the development of locomotor and reflex programs and in the context of structural heterogeneity of some of these muscles that is described in the companion paper. PMID- 8243576 TI - The distal hindlimb musculature of the cat: multiaxis moment arms at the ankle joint. AB - The cat hindlimb muscles have been classified, traditionally, as flexors and extensors, based on their actions in the parasagittal plane and their patterns of recruitment during locomotion and reflex responses. This study provides a detailed examination of the relative magnitudes of the various moment arms of the cat ankle muscles and the interdependent effects of position in the various axes of motion. We used a method based on observing small sliding movements of tendon in response to small angular displacements of the joint. Surprisingly, we found that the ankle joint of the cat permits substantial motion in three axes (eversion/inversion and abduction/adduction as well as extension/flexion) and many muscles crossing the ankle joint have their largest moment arms about axes other than extension/flexion. These moment arms often depended on the joint position in the axis of the moment arm and, to a lesser degree, on the extension/flexion angle as well. For some muscles (notably peroneus longus) there was sufficient variability that the predominant action in neutral posture (axis with the largest moment arm) could change from animal to animal, which may be related to heterogeneities of locomotor and reflex recruitment reported in the companion paper. PMID- 8243577 TI - The reach to grasp movement of blind subjects. AB - The importance of vision for the processing and coordination of the transport and manipulation components of a reach to grasp movement was assessed. Four blind volunteers (two men, two women; aged 25-40) were compared with matched control groups: (1) blindfolded and (2) full vision. Subjects reached 20 or 30 cm for a large or small diameter (6 cm or 0.7 cm, respectively) cylinder. For condition 1 trials they were given no instruction as to the type of grasp to adopt; for condition 2 they were instructed to consistently use a precision grip; while for condition 3 they were required to use whole had prehension. Blind subjects demonstrated a double grip pattern and either a low-velocity phase (20 cm) or a double transport movement (30 cm). However, their pattern of prehension with respect to intrinsic (size) and extrinsic (distance) cylinder properties was similar to that of the control groups. Grip aperture was appropriately scaled and, when greater precision was required, deceleration time was prolonged. Temporal coupling was evident between the two components. It was concluded that experience of vision is not necessary for the coordination or patterning of the basic reach to grasp movement. It does allow, however, for a movement consisting of only one opening and closing of the hand. PMID- 8243578 TI - The influence of stimulus orientation on the vertex positive scalp potential evoked by faces. AB - The scalp-recorded "vertex-positive peak" (VPP) evoked by images of faces in humans has previously been shown to be delayed when an originally upright stimulus is inverted or rotated by 90 degrees (Jeffreys 1989a). This paper describes a study of the effects on this scalp potential of smaller face orientation changes (15 degrees increments). The results showed that, under normal viewing conditions of clearly defined facial images, the VPP latency, which was minimal for face orientations within 15 degrees of the vertical (0 degree), increased almost linearly for incremental rotations from 15 up to 90 degrees, but was relatively unchanged or decreased slightly for further rotations from 90 up to 180 degrees. Similar results were observed for clockwise and anticlockwise rotations, and for different facial representations. These stimulus orientation changes did not change the latency of simultaneously recorded, pattern-specific potentials recorded from occipital scalp locations; nor did they greatly affect the VPP amplitude. By contrast, rotations of "Mooney figure" stimuli away from the vertical produced concurrent reductions in both the perception of a face and the amplitude of the evoked VPP. Experiments in which the orientation of both the stimulus face and the subject's head were varied further showed that minimal latency VPP responses were evoked for parallel stimulus and viewing orientations. The speed of response is thus determined by the orientation of the subtended retinal image. PMID- 8243579 TI - Ocular limit cycles induced by delayed retinal feedback. AB - Lisberger's and Robinson's models of smooth pursuit predict very different results from altering retinal feedback delay. We have therefore investigated the effects of increasing the retinal feedback time delay in three normal human subjects by means of an artificial feedback paradigm. When additional delays were incorporated into the retinal feedback path a threshold was reached beyond which the eye exhibited sustained self-excited oscillations or "limit cycles". The oscillation period increased linearly (as the added delay was increased) with slopes ranging from 1.41 to 1.6 with zero-delay intercepts of between 212 and 306 ms. Contrary to our experimental findings the Robinson and Lisberger models predict that the plot of oscillation period against added delay should have a slope of 3.4 and 2.7 and an intercept of 479 and 554 ms, respectively. Neither model produced comparable limit cycles, both being unstable at delays greater than 280 ms. Our results imply that the models of smooth pursuit need to incorporate predictive control. PMID- 8243580 TI - A quantitative analysis of the microglial cell reaction in central primary sensory projection territories following peripheral nerve injury in the adult rat. AB - The time course of the microglial cell reaction in central nervous system primary sensory projection territories has been examined following peripheral nerve injury in the adult rat using qualitative and quantitative analysis of immunoreactivity with the monoclonal antibody OX-42, which recognises the complement receptor CR3. The regions examined included the gracile nucleus, the column of Clarke and the spinal cord dorsal horn (superficial and deep laminae separately) after unilateral sciatic nerve transection, and the spinal trigeminal nucleus following unilateral infraorbital nerve transection. In all territories examined a qualitative increase in OX-42 immunoreactivity was observed 24 h postlesion. Further, quantitative analysis revealed an exponential development of the OX-42 immunoreactivity, with a peak at one week postlesion, thereafter showing a slow exponential decline. Our results show that the signal (or signals) that induces the microglial cell response in primary sensory projection territories is rapid in comparison to previously described central degenerative changes following peripheral nerve lesions (transganglionic degeneration). These findings are compatible with the hypothesis that activated microglia play a pathogenetic role in the development of transganglionic degeneration. PMID- 8243581 TI - Investigation of long-term recognition and association memory in unit responses from inferotemporal cortex. AB - We investigated recognition and association memory in the responses of single units isolated in the inferior temporal cortex of a macaque while it performed a visual discrimination task. The unit responses showed significant recognition memory (a decreased response upon image repetition). Furthermore, a recognition memory appeared to be a permanent feature in these units. Such memory was evident in responses recorded at least 1 h after the most recent presentations of the more familiar images and may have been built up over the months of training. For these cells, the shorter-term recognition memory (seconds) and the longer-term recognition memory (hour plus) were significantly correlated (0.68). In these same cells associative memory was investigated with ten abstract images which had been randomly and permanently paired. The monkey had been taught to discriminate these five pairs from other similar pairs of images. Neither the spike count nor temporal response shape (as determined by a principal-components analysis) showed increased similarity for the images that had been paired. The cells that had both short-term and long-term recognition memory had responses to previously paired stimuli that were no more similar than expected by chance. PMID- 8243582 TI - Botulinum toxin paralysis of the orbicularis oculi muscle. Types and time course of alterations in muscle structure, physiology and lid kinematics. AB - In chronically prepared guinea pigs, we investigated the time course of botulinum toxin A's (Bot A) effect on the blink reflex by monitoring lid movements and EMG activity prior to and after Bot A injection into the orbicularis oculi muscle (OOemg), or after nerve crush of the zygomatic nerve. We correlated these alterations with the morphological changes of the orbicularis oculi (lid-closing) muscles of the same animals. After Bot A treatment there was a profound reduction of OOemg activity and blink amplitudes as well as a slowing of maximum blink down phase velocity. Blink up-phases, however, remained unchanged. Gradual recovery of OOemg magnitude and blink amplitude started around day 6; a functioning blink reflex appeared on day 21, and full recovery of blink amplitude occurred by day 42. Crushing the zygomatic branch of the facial nerve produced similar changes in blink parameters, but recovery was much more rapid (15 days) than for Bot A treated guinea pigs. The morphological analysis demonstrated that Bot A produced a denervation-like atrophy in the orbicularis oculi. No fiber type-specific alterations were noted, and all muscle fiber types ultimately recovered, with no longstanding consequences of the transient denervation. Our findings support the notion that functional recovery was the result of preterminal and terminal axonal sprouting that subsequently re-established functional innervation. Moreover, differences between the present findings and those seen after injection of Bot A into the extraocular muscles strongly support the hypothesis that the composition in terms of muscle fiber type and the properties of the motor control system of a given muscle greatly influence both how the particular muscle responds to toxin injection, and how effective the toxin is in resolution of neuromuscular disorders that affect a particular muscle. The present findings were consistent with clinical observations that Bot A produces only temporary relief in patients with essential blepharospasm. It is likely that the efficacy of Bot A in treatment of blepharospasm could be improved by using agents that suppress terminal sprouting. The close correspondence of the changes in blink physiology between human patients and guinea pigs after Bot A treatment demonstrate that the guinea pig is an excellent model system for testing strategies to prolong the beneficial effects of Bot A treatment in relieving lid spasms in human subjects. PMID- 8243583 TI - The role of compensatory eye and head movements in the rat for image stabilization and gaze orientation. AB - Compensatory horizontal eye movements of head restrained rats were compared with compensatory horizontal eye-head movements of partially restrained rats (head movements limited to the horizontal plane). Responses were evoked by constant velocity optokinetic and vestibular stimuli (10-60 degrees/s) and recorded with search coils in a rotating magnetic field. Velocity and position components of eye and head responses were analysed. The velocity gains of optokinetic and vestibular responses of partially restrained and of head restrained rats were similarly high (between 0.8 and 1.0). Eye movements in partially restrained rats also contributed most (about 80%) to the velocity components of the responses. At stimulus velocities above 10 degrees/s, the "beating field" of the evoked optokinetic and vestibular nystagmus was shifted transiently in the direction of ocular quick phases. The amplitude of this shift of the line of sight was about 3 10 degrees in head restrained and about 20-30 degrees in partially head restrained rats. Most of this large, transient gaze shift (about 80%) was accomplished by head movements. We interpret this gaze shift as an orienting response, and conclude that the recruitment of the ocular and the neck motor systems can be independent and task specific: head movements are primarily used to orient eye, ear and nose towards a sector of particular relevance, whereas eye movements provide the higher frequency dynamics for image stabilization and vergence movements. PMID- 8243584 TI - Slow correcting eye movements of head-fixed, trained cats toward stationary targets. AB - Inspection of eye saccades made by head-fixed, trained cats revealed the existence of many eye shifts at an approximately constant velocity during the deceleratory phase of the saccade or at the end of it. Slow eye movements occurring at the end of a saccade are usually referred to as "postsaccadic drifts". It is shown that the duration and mean velocity of these "drifts" are related to the amplitude of the movement. The kinematics of these slow eye movements are nevertheless different from those of saccades. Slow movements at the end of the gaze shift have longer durations than those occurring during the intersaccadic interval between a saccade and a reacceleration of the eye. A closer study of the drifts of three trained cats showed that they play an important corrective role in reducing the residual error at the end of a saccade or during an intersaccadic interval. This functional corrective role was demonstrated by relating the amplitude of the slow movement to the amplitude of the residual error when the slow velocity eye shift began. It is therefore proposed to name these eye shifts "slow correcting movements". PMID- 8243585 TI - Types and time course of the alterations induced in monkey blink movements by botulinum toxin. AB - The alterations induced in eyelid movement metrics subsequent to unilateral injections of botulinum toxin type A into the orbicularis oculi muscle were studied in chronic alert monkeys using the search coil technique. Botulinum toxin caused rapid paralysis of blinks in the treated eyelid. The amplitude and peak velocity of blinks generated by this eyelid remained at or below 20% of that of the fellow, untreated eyelid for 10-20 days. Blink amplitude gain increased linearly thereafter, attaining control values by 40-60 days after injection. Recovery of blink peak velocity was slower. The adaptive alterations in blink duration that were observed during the acute phase of toxin paralysis suggest that the mechanisms responsible for blink reflex plasticity may produce bilateral adjustments in eyelid function. Taken together, these data establish a quantitative data base that can be exploited in order to: (1) better understand the neural adaptive mechanisms that operate during eyelid movements and (2) allow quantitative comparisons between current treatment protocols that employ botulinum toxin and protocols that may lead to improvements in the treatment of chronic eyelid spasm (blepharospasm). PMID- 8243586 TI - Structures mediating cholinergic reticular facilitation of cortical responses in the cat: effects of lesions in immunocytochemically characterized projections. AB - Cholinergic afferents to the neocortex controlled by the mesencephalic reticular formation (MRF) are known to transiently facilitate cortical excitability. In an attempt to identify the pathway mediating this effect in the cat visual cortex we combined retrograde tracing techniques with immunocytochemical methods to visualize the acetylcholine-synthesizing enzyme choline acetyltransferase (ChAT). In addition we examined, in acute electrophysiological experiments, whether local neurotoxin injections into nuclei of the basal forebrain interfered with the reticular facilitation of cortical evoked potentials. Cholinergic projections to area 17 originate from different centers in the homolateral substantia innominata/internal capsule, the septal nuclei, and the nuclei of the diagonal band of Broca. No direct cholinergic projection from the MRF to the visual cortex was observed. Retrogradely labelled cells intermingled with ChAT-positive neurons in the brainstem generally revealed immunopositivity for catecholaminergic markers. Local injections of neurotoxins in the substantia innominata blocked reticular facilitation, whereas local lesions of the septal nuclei and the nuclei of the diagonal band had no effect on MRF-induced facilitation. The blockage of the reticular facilitation of cortical evoked responses after unilateral lesions of the substantia innominata was bilateral, suggesting a cooperative interaction between basal forebrain structures of the two hemispheres. The anatomical and physiological data are discussed with respect to possible mechanisms of transient brainstem influences on cortical excitability. PMID- 8243587 TI - Nicotinic cholinoceptor-mediated excitatory postsynaptic potentials in rat nucleus ambiguus. AB - In rat brainstem slice preparations, intracellular recording from neurons (n = 39) in the compact formation of the nucleus ambiguus (AMBc) revealed spontaneous and miniature excitatory postsynaptic potentials (EPSPs; n = 11) that, along with acetylcholine-induced depolarization, were enhanced by physostigmine (10 microM; n = 2) and blocked by dihydro-beta-erythroidine 1-5 pmol (n = 4). Retrograde neuronal tracing combined with choline acetyltransferase immunocytochemistry demonstrated that the AMBc receives a projection from a subpopulation of cholinergic neurons in the zona intermedialis reticularis parvicellularis. Electrical stimulation of this region in slices evoked fast EPSPs in AMBc neurons (n = 23) that were inhibited by dihydro-beta-erythroidine 2-5 pmol (n = 8), but not by methscopolamine 1 pmol (n = 2). The present findings strongly support the existence of a cholinergic nicotinic synapse mediating fast transmission in brainstem vagal motoneurons. PMID- 8243588 TI - Evidence for a preventive action of the vigilance-promoting drug modafinil against striatal ischemic injury induced by endothelin-1 in the rat. AB - We have studied the ability of the vigilance-promoting drug modafinil to counteract the ischemic lesion produced by a unilateral microinjection of endothelin-1 (ET-1) in the neostriatum of the rat using a combined morphometrical, biochemical, cardiovascular and behavioral analysis. ET-1 was injected unilaterally into the neostriatum. The ET-1-induced lesion volume, which was determined by a computer-assisted morphometrical analysis, was reduced by the 7-day modafinil treatment (10, 30, and 100 mg/kg i.p.) in a dose-related way. Modafinil also produced a dose-related counteraction of the ET-1-induced increase of perfusate lactate levels, as determined by intrastriatal microdialysis without affecting the ET-1 induced reduction of striatal blood flow, as determined by laser-Doppler flowmetry. The ipsilateral rotational behavior induced by apomorphine in the ET-1-lesioned rats was reduced dose-dependently by modafinil treatment. Thus, morphological, neurochemical, and behavioral evidence that the putative ischemic striatal injury induced by microinjection of ET-1 in the rat neostriatum is counteracted in a dose-dependent way by modafinil treatment has been obtained. The mechanism does not appear to involve an increase in striatal blood flow. It is instead speculated that its powerful preventive action in striatal ischemic injury may be related to a reduced anaerobic metabolism. PMID- 8243589 TI - Etiology of intoxication: a study of 557 cases. AB - Our results were obtained from the emergency service at Elche General Hospital, which covers an adult population of 275,076 inhabitants. We studied intoxicated patients aged 15 years and above, during 1988. Our aim was to investigate the etiological and epidemiological circumstances of intoxications. We obtained a prevalence of voluntary cases (61.6%) over accidental cases (38.3%), with self poisoning being more frequent than drug addiction, in the first group and animal stings being more frequent than domestic or occupational intoxication in the second. Mortality resulted in 1.6% of cases; the etiology of the highest mortality was self-poisoning and the most common toxic substances were caustic substances. PMID- 8243590 TI - Studies on the epidemiology of child infections in the Bari area (south Italy). VII. Epidemiology of Epstein-Barr virus infections. AB - Serological patterns against Epstein-Barr virus (EBV) specific antigens were determined in 3732 healthy babies and children aged 0-10 years living in the Bari area (South Italy). IgG antibodies against EBV capsid antigen (VCA) were found in 2713 subjects (72.7%). Seropositivity rates, high in the first semester of life (83.8%), declined between 6 and 12 months (65.6%) and even further between 1 and 2 years (43.8%). After 2 years the frequency of positive children rose progressively reaching steady levels between 5 and 7 years (80.2%) and between 8 and 10 years (81.9%). IgA antibodies against VCA, IgG anti-early viral antigen (EA) and IgG against virus-associated nuclear antigens (EBNA) were found in 17.9%, 15.9% and 25.7% of the subjects tested, respectively. IgM anti-VCA were found only in 35 (0.9%) children, but 818 (21.9%) exhibited antibody patterns suggestive of a recent infection: IgG anti-VCA > or = 1:160 alone or in association with IgA anti-VCA or IgG anti-EA or both. These results suggest that in this area the primary infection by EBV occur early in life, with immunity to EBV acquired primarily after 4 years. PMID- 8243591 TI - Prevalence of congenital abnormalities in Turkish school children. AB - 19750 school children, ages 6 to 15 years, were examined by the authors of this study, 1,220 (6.18%) had congenital abnormalities. In this group, 4.23% were boys and 1.88% were girls. Case histories revealed inbreeding amongst the parents (families) of children with congenital malformation to be 8.9% and 8.2% for the rest of the families in this study. There were 27 different congenital abnormalities identified, with prevalence rates of 0.05/1,000 to 15.85/1,000. The most prevalent abnormalities were umbilical hernia (15.85/1000), inguinal hernia (14.50/1,000), pectus carinatum and excavatum (7.68/1,000), undescended testes (9.00/1,000 boys), congenital nevus (3.54/1,000), retractile testis (4.45/1,000 boys), pilonidal sinus (2.63/1,000), pes planus (2.28/1,000), and hemangioma (1.16/1,000). Of the 19,750 children, 70 had multiple anomalies (3.75/1,000). PMID- 8243592 TI - Epidemiology of cardiovascular disease risk factors in Catalonia (Spain). AB - Cardiovascular disease is one of the most important public health problems in developed countries. We have studied the epidemiology of the following cardiovascular disease risk factors in a random sample (n = 704) of the adult population of Catalonia (Spain): hypercholesterolemia (> or = 6.1) mmol/l or 240 mg/dl), hypertension (SBP > or = 160 and/or DBP > or = 95 mmHg), low HDL cholesterol concentrations (< 0.9 mmol/l or 35 mg/dl), hypertriglyceridemia (> 2.8 mmol/l or 250 mg/dl), obesity (BMI > 30), smoking and history of diabetes and coronary heart disease. Two percent of participants had hypertriglyceridemia, 3% had a history of coronary heart disease, 4% a history of diabetes, 6% low HDL cholesterol concentrations, 12% were obese, 20% had hypertension, 24% had hypercholesterolemia and 36% were smokers. 58% of hypertensive individuals had been previously detected, 46% were currently on treatment, and 21% had their blood pressure controlled (SBP < 160 and DBP < 95 mmHg). Correlation and multiple regression analyses were used to investigate the association between cardiovascular risk factors. Multiple linear regression analysis showed independent correlations between risk factors. Prevalence of hypercholesterolemia, obesity and diabetes was higher and prevalence of smoking was lower in hypertensives than normotensives. The odds ratio was 3.68 (95% CI = 2.07-6.54) for hypercholesterolemia, 3.26 (95% CI = 1.52-7.02) for obesity, 3.81 (95% CI = 1.09-7.02) for diabetes and 0.40 (95% CI = 0.22-0.70) for smoking. The adjusted odds ratio was statistically significant for hypercholesterolemia (OR = 2.74, 95% CI = 1.01-3.75). The prevalence of cardiovascular risk factors was similar to that observed in other Mediterranean communities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243593 TI - Nosocomial outbreak of severe Pseudomonas aeruginosa infections in haematological patients. AB - From June to September 1988, an outbreak of Pseudomonas aeruginosa infections in neutropenic patients admitted to the Haematological Wards of "Ospedali Riuniti" in Bergamo, Italy, was detected. Out of 11 cases of P. aeruginosa infections, 8 were bacteremic. Of these, 7 died within few days of onset (mortality rate: 87.5%). Consequently, possible sources of infection were investigated, and moist areas of the hospital environment were shown to be highly contaminated by P. aeruginosa. A clinical and microbiological follow-up of patients admitted to the Haematological Wards was performed for a 10 month period following the outbreak. Adequate measures for cleaning and disinfection were shown to reduce the frequency of P. aeruginosa hospital infections. PMID- 8243594 TI - The Ixodes ricinus tick as a vector of Borrelia burgdorferi in Slovenia. AB - In many parts of Slovenia, the vegetation, climate and animals form an ideal natural environment for the persistence of different tick species. In this study, Ixodes ricinus ticks, the prevalent tick species in our country, are identified as the vectors of Borrelia burgdorferi in Slovenia. Ticks were collected by flagging between May and October 1990. All the ticks collected from different parts of Slovenia were I. ricinus and each one was examined individually. The ticks were tested to determine the presence of B. burgdorferi by indirect immunofluorescent assay. Among the 496 ticks examined for borreliae there were 85 adults and 411 nymphs. Twenty (23.5%) adults and 18 (4.4%) nymphs were found positive for Borreliae. This study discusses the importance of I. ricinus ticks in the role of Lyme borreliosis in Slovenia. PMID- 8243595 TI - Echographic epidemiology of cystic hydatid disease in Italy. AB - This study was carried out in order to establish the actual prevalence of cystic hydatid disease in Italy using ultrasound. In fact, there is a great difference between the official reports of the disease and epidemiological investigations. From 1988 to 1990, 424 cases of cystic hydatid disease were diagnosed in 333,144 examinations performed in nineteen Italian Echographic Departments, giving a mean incidence of 0.16%. Most of the examined persons arriving at the 19 Echographic Departments had symptoms independent from hydatidosis, were affected with other pathologies, or they were patients attending follow-up for neoplasms. The authors note that human hydatidosis has a high incidence in the south and in the isles; hepatic lesions were 85.61% of all cases of abdominal hydatidosis, asymptomatic cystic hydatid disease was present in 60% of the total number; the average age of the sample examined was 45.38 years without significant differences between men and women. Finally the Authors suggest the importance of echography in planning therapeutic programs. PMID- 8243596 TI - Asymptomatic pharyngeal carriage of beta-haemolytic streptococci and streptococcal pharyngitis among patients at an urban hospital in Croatia. AB - In a one-year study at the University Hospital of Infectious Diseases in Zagreb, Croatia the rate of asymptomatic throat carriage of beta-haemolytic streptococci was investigated. Throat carriage was investigated in 1796 patients, none of whom had a sore throat nor signs and/or symptoms of a respiratory tract infection. The carrier rate of beta-haemolytic streptococci was 8.3%, for group A streptococci 6%, group B 1.3%, group C 0.3% and group F 0.1%. The highest rate was observed in the 6 to 14 year age group: 13.8% for all streptococcal groups, 11.7% for group A alone. The proportion of non-A streptococci was higher in older age groups. Tonsillectomised individuals were less frequently carriers. No sex or season dependent variations were observed. In a four-month study of 629 patients with pharyngitis the throat cultures yielded: group A streptococci in 44.7%, group B in 1.7%, group C in 0.8%, and group G in 0.6% of the patients. Group A streptococci in 44.7%, group B in 1.7%, group C in 0.8%, and group G in 0.6% of the patients. Group A streptococcal rates in carriers compared to rates in patients with pharyngitis suggest that approximately one fourth of the schoolchildren with culture-positive pharyngitis actually are not truly infected, but only carriers currently having a sore throat of non-streptococcal etiology. Antibiotics were administrated to 605 (96.2%) patients with pharyngitis; in 571 (94.4%) of the cases before culture results became available. After culture negative results were obtained the therapy was discontinued in only 28.5% of the cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243597 TI - Plasmid based differentiation and detection of Coxiella burnetii in clinical samples. AB - A "nested" PCR approach with primers based on conserved plasmid sequences was used for the highly sensitive and specific detection of Coxiella (C.) burnetii in clinical samples collected from cattle, dogs, cats and humans. Results were in good agreement with those obtained from Capture-ELISA and isolation of the organism in BGM cell culture. We also tested primers with sequences derived from genomic DNA and sequences based on 16S rRNA. In addition, we applied PCR for the differentiation of C. burnetii plasmid types from 28 isolates originating from the USA, Europe and South Africa. Reference isolates Nine Mile RSA493, Dugway 5J108-111 and all European isolates tested were recognized only by primers specific for the QpH1 plasmid. One isolate from a goat abortion in Namibia reacted identically to the reference isolate Priscilla Q177 bearing the QpRS plasmid. Reference isolate S Q217 with plasmid sequences integrated into the genome reacted with none of the plasmid-specific primer pairs. PMID- 8243598 TI - Diagnosis of HIV-1 infection by PCR with two primer pairs. AB - We have used two different primer pairs to assess HIV-1 infection of peripheral blood mononuclear cells (PBMC) by polymerase chain reaction (PCR). The study was carried out on 150 individuals: 50 seronegative individuals without risk behaviours for HIV-1 infection, 50 individuals with risk-behaviours but seronegative for HIV-1 and 50 patients with risk-behaviours who were HIV-1 seropositive. Discordances were found between the two primer pairs (SK38/39; SK68/69) in 3 cases. In the non-risk seronegative group, one specimen was scored positive with only one primer pair (SK38/SK39); all the samples belonging to seropositive individuals were found to be positive for HIV-1 DNA using both primer pairs; in the seronegative at risk group 2 samples were positive with only one primer pair (SK38/SK39), and 4 samples were found positive by both primer pairs (SK38/39 & SK68/69). Our study demonstrates that discrepant results take place with relatively high frequency; we propose that all specimens should be tested twice using at least two different primer pairs. PMID- 8243599 TI - Anti-HIV 1 antibodies prevalence in parturients through newborn testing: results of the Italian anonymous serosurvey. The Italian Collaborative Study Group of HIV Prevalence in Newborns. AB - A prevalence assessment of HIV infection among parturients was performed in order to estimate the future incidence of pediatric AIDS cases and obtain data on the trend of the infection in Italy. Consecutive whole blood samples from newborns collected on filter paper from all regions of Italy (October-December 1990) for routine metabolic screenings, were anonymously examined for HIV-1 antibodies by an ELISA technique. Positive results were confirmed by Western blot. Among 97,658 blood samples tested, 121 (0.124%, 95% confidence interval Poisson distribution 0.103-0.148) were positive. A high prevalence of HIV infection (> 0.2%) was observed in four regions from North and Central Italy while, in some regions in the South, the seroprevalence was ten times lower. In the Lazio Region, namely in Rome, 5 inner-city hospitals with a particularly high prevalence of HIV infection were identified. Assuming that the prevalence of HIV infection observed in the population in this study may reflect the infection rate in the total population of childbearing women, we estimate that approximately 700 newborn babies were delivered from HIV-positive mothers in Italy during 1990. The identification of high prevalence areas could allow for the development of "targeted" testing programs to plan and provide adequate counselling and care of HIV-infected women and their infants. PMID- 8243600 TI - Estimation of the magnitude of the HIV epidemic among homosexual men: utilization of survey data in predictive models. AB - A group of 709 randomly selected males aged 18-55 years living in the city of Amsterdam, the Netherlands, were questioned about sexual lifestyle and related behavior. A substantial proportion (10%) of these men appeared to be homo- or bisexual. Of these homo- and bisexual men, 16.5% were predicted to be HIV seropositive, using their behavioral information in logistic regression models derived from a second study among 506 homo- or bisexual volunteers who were questioned and tested for HIV antibodies as well. Generalizing the results of the sample, the magnitude of the homo- and bisexual population in Amsterdam is estimated to be 21,500 (95% confidence interval: 17,000-26,000) of whom 3500 are HIV seropositive. This is in agreement with estimations on the basis of AIDS surveillance data. The study shows that in the absence of serological data it is possible to estimate HIV prevalence, given that the relation between the presence of HIV antibodies and sexual activity or indicators for sexual activity is known. Since serological data are lacking in many occasions, this method may be of help to estimate HIV prevalence in other geographic locations and risk groups. PMID- 8243601 TI - Predictive equation for acquisition of hepatitis B in hospital workers in a general hospital. AB - A cross-sectional study was performed to obtain risk factors for hepatitis B disease, HBsAg carriers and immunised personnel, among 2470 workers in a general hospital in Madrid, Spain. The data obtained were analyzed with multiple logistic regression to obtain beta coefficients for variables. The results of the analysis show that being a nurse or being regularly exposed to blood are the most important risk factors for hepatitis B acquisition. The length of time working at the same job activity was also a risk factor. The resulting beta coefficients allow the construction for a hepatitis non-immunised, HBsAg carrier and immunised HBV status, which can select subjects for a hepatitis B vaccination program. PMID- 8243602 TI - Simultaneous infection with three different S. enteritidis strains in a nursing home resident. AB - A culture taken from a nursing home resident as part of a S. enteritidis outbreak was found to have a mixed infection due to three different strains of S. enteritidis. One of the three strains belonged to phage type (PT) 4, one to PT6 and one reacted with phages but did not conform to any typing scheme (RDNC). All three strains had the 38.9 megadaltons (MDa) plasmid found in the isolates from the outbreak-related cases, in addition the PT6 and RDNC strains harboured a 69.9 MDa plasmid. The importance of phage typing and plasmid analysis for S. enteritidis strain characterization and their epidemiologic and bacterial significance are discussed. PMID- 8243603 TI - Evaluation of rapid urease test for detection of Helicobacter pylori in gastric biopsy specimens. AB - Using 80 gastric biopsy specimens from patients with various gastroduodenal diseases, the isolation rate of H. pylori and urease activity of the biopsy specimens were examined. The sensitivity and specificity of the rapid urease test was 84.6% and 75.9%, respectively, for the cultivation of H. pylori. The mean number of H. pylori detected in urease-positive and negative biopsy specimens was 10(6.1 +/- 0.9) and 10(4.5 +/- 1.2) cfu/g specimens, respectively. PMID- 8243604 TI - A Cyanobacterium-like body found in the stools of an HIV+ patient with diarrhoea. PMID- 8243605 TI - Emergence of oropharyngeal candidiasis caused by non-albicans species of Candida in HIV-infected patients. PMID- 8243606 TI - Human neutrophil granules and secretory vesicles. AB - The traditional classification of neutrophil granules as peroxidase-positive (azurophil, or primary) and peroxidase-negative (specific or secondary) has proven to be too simple to explain the differential exocytosis of granule proteins and incorporation of granule membrane into the plasma membrane which is an important aspect of neutrophil activation. Combined subcellular fractionation and immunoelectron microscopy has revealed heterogeneity among both peroxidase positive and peroxidase-negative granules with regard to their content, mobilization and time of formation. Peroxidase-negative granules may be classified according to their content of lactoferrin and gelatinase: 15% of peroxidase-negative granules contain lactoferrin, but no gelatinase. 60% contain both lactoferrin and gelatinase. The term specific or secondary granule should be reserved for these two subsets. In addition, 25% of peroxidase-negative granules contain gelatinase but no lactoferrin. These should be termed gelatinase granules or tertiary granules. Gelatinase granules are formed later than specific granules and mobilized more readily. In addition, a distinct, highly mobilizable intracellular compartment, the secretory vesicle, has now been recognized as an important store of surface membrane-bound receptors. This compartment is formed in band cells and segmented cells by endocytosis. This heterogeneity among the neutrophil granules is of functional significance, and may also be reflected in the dysmaturation which is an important feature of myeloproliferative and myelodysplastic disorders. PMID- 8243607 TI - Heart transplantation in a case of juvenile hereditary haemochromatosis followed up by MRI and endomyocardial biopsies. AB - Cardiac involvement in hereditary haemochromatosis (HH) is a poor prognostic sign and is the main cause of death in the juvenile form. The treatment of choice is iron removal therapy by phlebotomy, but treatment by iron chelation (desferrioxamine) has been recommended in cases with severe cardiac symptoms. We describe here the first case of juvenile HH undergoing heart transplantation, which became necessary despite intensive iron removal therapy by phlebotomy and treatment by desferrioxamine. Throughout the course the myocardial iron content was monitored by endomyocardial biopsies and by magnetic resonance imaging (MRI). At the last follow-up, 18 months after transplantation, the myocardial iron content in the transplanted heart was still within reference ranges by biochemical determination and MRI and the patient's condition was completely satisfactory. In conclusion, heart transplantation should be considered in cases of severe juvenile HH. In the follow-up of these patients MRI may be a useful supplement. PMID- 8243608 TI - Therapeutic potential of intravenous 67-gallium in non-Hodgkin's lymphoma. AB - 67-gallium accumulates rather selectively in malignant lymphoid tissue. The isotope has a substantial cytotoxic effect in human-derived cell-lines. 67 gallium was given intravenously to 3 patients with end-stage, resistant large cell lymphoma. Evaluation of tumour response was done by physical measurements, and CT-scanning together with gallium scintigraphy. Three weekly doses of 20, 40 and 60 mCi respectively caused persistent pancytopenia in 1 patient. Pancytopenia was not observed in 2 other patients given two 40 mCi doses 4 weeks apart. In all 3 patients, some response was noted, be it shortlived and different from site to site. 67-gallium has some cytostatic effect in large cell lymphoma. It seems feasible to start a phase I study to find a tolerable dose to be given every 4 weeks. PMID- 8243609 TI - Synergistic cytotoxicity of AZT plus alpha and gamma interferon in chronic myeloid leukemia cell line K562. AB - We have previously reported that the antineoplastic activity of 3'-azido 3' deoxythymidine (AZT) can be increased by drugs that inhibit "de novo" thymidylate synthesis, such as 5-fluorouracil, methotrexate and hydroxyurea. In the present study we tested the combinations AZT+alpha interferon (IFN) and AZT+gamma IFN on in vitro growth of the human acute-phase chronic myeloid leukemia (CML) cell line K562. After 72 hours incubation, not only AZT+alpha-IFN but also AZT+gamma-IFN were synergistic in inhibiting K562 growth, as demonstrated by isobologram analysis of the data. This enhanced cytotoxicity was confirmed by the evaluation of [3H]AZT incorporation into cellular DNA, that was increased by 50% and 222% in the presence of alpha- and gamma-IFN, respectively. The addition of 50 mumol/l thymidine to the culture medium was able to reduce the cytotoxicity of the drug combinations to the degree observed with each compound alone; furthermore, the increased incorporation of AZT into DNA was completely reversed. These data indicate the existence of a biochemical interaction between AZT and IFNs that results in an increased cytotoxic effect. While the combination AZT+alpha-IFN is currently being tested in HIV-related malignancies, AZT+gamma-IFN is new and deserves further study in human CML acute and chronic phase models, in view of possible clinical applications. PMID- 8243610 TI - B-chronic lymphocytic leukaemia in Latvia: epidemiological aspects. AB - As in western Europe and the USA, chronic lymphocytic leukaemia (CLL) in Latvia is the most prevalent type of leukaemia. A total of 1509 newly diagnosed cases of B-cell chronic lymphocytic leukaemia entered the study, 440 of whom were followed up at the Latvian Haematology Centre. The main peculiarities of the study were: 1) the higher incidence of the disease in Latvia as compared with other regions of the former USSR and with western countries, 2) a lower male-to-female ratio than in most other countries and 3) a higher morbidity among the Latvian population in comparison with the Russian. A positive correlation between the disease stage and survival was confirmed, but no precise individual prognosis in an early stage of the disease was possible. PMID- 8243611 TI - Production of oxygen radicals by fibroblasts and neutrophils from a patient with x-linked chronic granulomatous disease. AB - Recently, a superoxide-generating NADPH-oxidase system in human fibroblasts has been described. Therefore, we reassessed the possible use of this cell type for prenatal diagnosis of CGD patients comparing normal and CGD peripheral blood neutrophils (PMN) and skin fibroblasts in their reactive oxygen intermediate (ROI)-producing capacity. While PMN of the CGD patient showed a clearly reduced respiratory burst activity, which correlated well with the measured content of cytochrome b558, fibroblasts of the same individual showed no impaired production of superoxide anion or H2O2 upon stimulation by cytokines (TNF and IL-1) or other agents (Ca2+ ionophores and PAF, unpublished results). Furthermore, fibroblasts of the CGD patient or of normal donors could be inhibited in ROI production by diphenylene iodonium (DPI) and 2-iodobiphenyl. In contrast to PMN, no inhibition of the fibroblast NADPH-oxidase system was observed using staurosporin, an inhibitor of proteinkinase C. These data demonstrate, in contrast to previous studies, that fibroblasts are able to produce ROI. Nevertheless, since fibroblasts obtained from a CGD patient exhibited no difference in ROI production compared with fibroblasts obtained from healthy donors, they are not suitable for prenatal diagnosis of CGD. PMID- 8243612 TI - Primary polycythaemia: diagnosis by non-conventional positive criteria. AB - Patients with polycythaemia and normal controls have been studied to establish and subsequently test non-conventional criteria for the diagnosis of primary polycythaemia (primary proliferative polycythaemia, polycythaemia vera) as compared with conventional Polycythaemia Vera Study Group (PVSG) assessment. One criterion was erythroid colony formation from peripheral blood in a serum-free system, assayed alone and with the addition of recombinant human erythropoietin (Epo), interleukin 3 (IL3), or alpha interferon (alpha-IFN) (Dudley et al. 1990). The remaining criteria were non-culture associated and comprised platelet distribution width (PDW), platelet nucleotide ratio (ATP:ADP), serum erythropoietin and clinical evidence of ischaemic vascular disease. The combination of culture associated and non-culture associated variables, by use of a simple additive scoring system, gave no false positive and only 6% false negative results in distinguishing primary polycythaemia from other polycythaemias and normal controls in those (34 patients Group A) used in its derivation. Testing the scoring system in a newly presenting group (25 patients Group B) was highly satisfactory with no false positives and only a few false negative results (14%). Use of these non-conventional criteria should allow more confident diagnosis of primary polycythaemia, where conventional clinical and laboratory assessment is inconclusive. PMID- 8243613 TI - Circulating megakaryocytes: delivery of large numbers of intact, mature megakaryocytes to the lungs. AB - To determine the locus of platelet production, we sought to determine if sufficient megakaryocytes reach the lungs in a state that could produce platelets. Elutriation was used to isolate megakaryocytes from blood reaching and leaving the lungs of 20 patients undergoing routine cardiac catheterizations. A mean of 5.0 intact megakaryocytes/ml were found in pulmonary artery blood, compared to only 0.5 megakaryocytes/ml, with partial cytoplasmic content, in aortic samples. The megakaryocytes in central venous and aortic samples were all mature. The identity of these cells as megakaryocytes, their maturity and normal morphology were confirmed by standard and immunoelectron microscopy. Cardiac outputs were obtained for each patient at the time of blood sampling, allowing an extrapolation that 40 x 10(6) intact, mature megakaryocytes were being delivered to the lungs every day in the average patient, compared to only 4.0 x 10(6) partially spent megakaryocytes exiting the lungs daily. About 98% of megakaryocyte cytoplasm reaching the lungs did not exit as recognizable megakaryocytes or fragments. The number and state of the megakaryocytes apparently filtered in the lungs is consistent with the hypothesis that megakaryocytes may shed platelets within the pulmonary microvasculature, which may be the primary site of platelet production. PMID- 8243614 TI - Pharmacokinetic dosing in prophylactic treatment of hemophilia A. AB - The aim of this study was to investigate individual pharmacokinetics as a tool for dosing of factor VIII (FVIII) in severe hemophilia A. It is assumed that effective prophylaxis against bleedings is maintained if the plasma FVIII:C activity is kept above 1 U/dl, and the present study is based on this assumption. A current standard dosage regimen for FVIII is 25-40 U/kg up to three times weekly. However, there is considerable individual variation in the pharmacokinetics of FVIII:C. Individual pharmacokinetic data were used to computer-simulate plasma activity curves after repeated doses in 8 patients. Going from prophylaxis regimens of normally 2-3 infusions per week to dosing every 2 days would theoretically reduce their average FVIII consumption by 43% with maintained or increased trough levels of FVIII:C. Daily dosing would reduce their mean FVIII usage by 82%. Modified dosage regimens, infusions every 2 days, were implemented in the patients, and plasma samples were drawn to verify the pharmacokinetic models. The feasibility of the method to generally raise trough levels with a decreased consumption of FVIII was confirmed. Dosing of coagulation factors according to kinetic principles can result in more cost-effective utilization of these very expensive preparations. PMID- 8243615 TI - Neuropeptide Y (NPY) as a tumor marker. PMID- 8243616 TI - The yield of bronchoalveolar lavage in the etiological diagnosis of pneumonia in leukemia and lymphoma patients. PMID- 8243617 TI - Beyond the "psychosomatic family": a biobehavioral family model of pediatric illness. AB - This article proposes a multilevel systems model of the interactive biological, psychological, and social processes in childhood illness. This heuristic model can organize the investigation of pathways and mechanisms by which these levels influence one another. A pivotal feature of this biobehavioral family model is the construct of responsivity, which is conceptualized at both family (interpersonal responsivity) and individual (biobehavioral reactivity) levels. Individual biobehavioral reactivity is proposed as a bidirectional pathway by which family patterns and disease processes influence one another. Illustrative clinical material and relevant research are presented with recommendations for future study. This model integrates individual and family-level theory and thus has the capacity to bridge the fields of psychosomatics, pediatrics, behavioral medicine, and family systems medicine. PMID- 8243618 TI - A case of functional urinary retention: the use of family play therapy. AB - This case presents a technique for doing family therapy in families with young children. In family play therapy, the entire family plays together, allowing full participation of even the youngest children in the therapeutic process. Standard family therapy interventions in the latter part of each session make use of family interaction patterns and unconscious processes revealed during the earlier play. In the case presented here, the technique was successfully applied to a family with a 3-year-old son who had functional urinary retention. This case also provided an opportunity for interdisciplinary collaboration between the separate Child and Family Services at a major teaching hospital. PMID- 8243619 TI - How problems evolve and dissolve: integrating narrative and strategic concepts. AB - This article presents an approach to therapy that links narrative and strategic concepts. The term "strategic" is used not in the prescriptive, impositional sense that has come to be associated with the method, but in terms of having a clear therapeutic direction in promoting change. The authors outline an approach to therapy that expands upon the fundamental principles of the MRI (Mental Research Institute) Brief Therapy model, elaborating more upon its constructivist premises than its prescriptive practices. They propose that by mapping how ordinary life events affect a person's preferred view, the therapist can locate the key narrative elements that shape the course of the problem and direct its solution. The authors suggest a framework for how problems evolve and dissolve. When new events are construed as contradicting family members' preferred narrative accounts, problems evolve. Problems dissolve when family members see the event, and the ideas and actions of others, as consonant with their preferred ways of being and acting. PMID- 8243620 TI - Couples therapy using the technique of construct differentiation. AB - A central problem in couples therapy is the clients' tendency to persist in endless and fruitless alternation between attacking the partner and justifying themselves. The technique of construct differentiation described in this article has proved helpful in coping with this situation. In three clearly structured steps, the therapist explores the personal constructs of each partner in the presence of the other. Instead of concentrating in the usual way on circularity of thoughts, feelings, and behavior, the partners are hindered from interacting and are confronted by the therapist's questions concerning their own inner process, independently of that of the partner. By becoming aware of their own ambivalent constructs and those of the partner, the eagerness to convert the partner to their own point of view can be reduced, and the construct system of each partner can be enriched by the other's constructs. Construct differentiation proves especially useful in the treatment of power struggles and narcissistic collusions. PMID- 8243621 TI - Missing links: the use of enthymemes and their applications for family therapists. AB - The enthymeme is a rhetorical concept that has been defined in a number of ways throughout the ages. A review of these definitions creates space for a broad enthymematic concept based on the various descriptions, rather than on a limiting, "definitive" definition. A number of examples are offered--from the work of Milton Erickson on hypnosis and from nonhypnotic therapy sessions--that demonstrate the presence and usefulness of enthymemes in family therapy dialogue. PMID- 8243622 TI - Maternal interaction style in affective disordered, physically ill, and normal women. AB - Affective style (AS) and communication deviance (CD) have been suggested as markers of dysfunctional family environments that may be associated with psychiatric illness. Studies have focused mainly on parental responses during family interactions when an offspring is the identified patient. The present study is unique in examining AS and CD in mothers with unipolar depression, bipolar disorder, or chronic physical illness, and in normal controls. The sample consisted of 64 mothers with children ages 8 to 16. Unipolar mothers were more likely to show negative AS than were any other maternal group. There were no group differences for CD. Chronic stress, few positive life events, and single parenting were associated with AS. CD was associated solely with lower socioeconomic status. Results suggest that dysfunctional interactions are determined not only by maternal psychopathology, but also by an array of contextual factors that are related to the quality of the family environment. PMID- 8243623 TI - Family transitions and developmental processes in panic-disordered patients. AB - Although there is increasing evidence for a familial role in panic disorder, little research exists on the family interaction patterns and relational stressors that may contribute to this disorder. Using DSM-III-R criteria, 52 families were classified into Panic Disorder, Major Depressive Disorder, and Nonclinical Control groups. A semi-structured interview and self-report measures were then used to examine family processes. MANOVA results were highly significant, suggesting that, compared to depressed and nonclinical control families, panic-disordered families had unresolved life-cycle issues, were enmeshed, used triangulation, and failed to resolve conflicts. These findings suggest that dysfunctional family processes may be involved in the expression and maintenance of panic disorder. PMID- 8243624 TI - Dropping out of marriage and family therapy: a critical review of research. AB - The lack of attention given to research on the premature termination by clients in marriage and family therapy is evident in research reviews of the dropout phenomenon in psychotherapy. This article is an attempt to fill that void. The research literature on dropping out of family therapy is reviewed and organized. Studies in this area tend to focus on four kinds of variables: client characteristics, therapist characteristics, therapy process variables, and interventions. Stable findings in each of these areas are pointed out. Both limitations and recommendations for future research are discussed. PMID- 8243625 TI - Bowen theory. PMID- 8243626 TI - Super-secondary structures involving triple-strand beta-sheets. AB - Triple-strand beta-sheets having up- and -down topology are widespread in proteins and occur in two forms denoted here as S-like and Z-like beta-sheets. In many cases they are included in super-secondary structures of higher order. A number of such structures is described in this paper. An important feature of these super-secondary structures is that they have a unique handedness. Another feature is that some of them only involve S-like beta-sheets and others only Z like beta-sheets. PMID- 8243627 TI - Selectivity of phospholipase C isozymes in growth factor signaling. AB - Xenopus laevis oocytes were injected with mRNA extracted from growth factor responsive CCL39, Chinese hamster lung fibroblasts. The expression of functional growth factor receptors on the oocytes was demonstrated by growth factor-induced 45Ca2+ efflux. To determine the isozyme(s) of phospholipase C (PLC) coupled to growth factor receptors, growth factor-induced 45Ca2+ efflux were measured following coinjection of mRNA from CCL39 cells with PLC antibodies. PLC-gamma 1 antibody did not lead to loss of 45Ca2+ efflux induced by thrombin but resulted in loss of that induced by platelet-derived growth factor (PDGF). In contrast, PLC-delta 1 antibody did not block PDGF-induced 45Ca2+ efflux but led to inhibition of thrombin-induced 45Ca2+ efflux. PLC-beta 1 antibody did not affect Ca2+ efflux by the treatment of either thrombin or PDGF. These results suggest that these growth factor receptors are coupled to specific effectors, i.e. thrombin receptor to PLC-delta 1 and PDGF receptor to PLC-gamma 1. PMID- 8243628 TI - Azidobutyryl clentiazem, a new photoactivatable diltiazem analog, labels benzothiazepine binding sites in the alpha 1 subunit of the skeletal muscle calcium channel. AB - [3H]Azidobutyryl clentiazem, a new photoactivable diltiazem derivative, has a higher binding affinity than azidobutyryl diltiazem. It can be covalently incorporated into the alpha 1 subunit of the skeletal muscle calcium channel by UV irradiation, which allows the benzothiazepine binding site to be determined. The photolabeled alpha 1 subunit and its proteolytic fragments were analyzed with a panel of sequence-directed antibodies. The results suggest that the linker region between segment S5 and S6 of domain IV is involved in benzothiazepine binding. This site is different from the dihydropyridine and verapamil binding sites. PMID- 8243629 TI - Secondary structure of globular proteins at the early and the final stages in protein folding. AB - The ellipticities for an early transient intermediate in refolding observed by kinetic circular dichroism measurements at 220-225 nm for 14 different proteins are summarized, and the ellipticity values are compared with those for the final native proteins and also with the ellipticities expected from a physical theory of protein and polypeptide secondary structure. The results show that a substantial part of the protein secondary structure is in general formed in the earliest detectable intermediate in refolding and that the ellipticities in both the native and the intermediate states are consistent with the physical theory of protein secondary structure. PMID- 8243630 TI - Interaction of clavulanate with class C beta-lactamases. AB - The interactions between clavulanate and three class C enzymes have been studied in detail. In all cases, the reactions followed branched pathways where 25-150 turnovers occurred before inactivation was completed. Reactivation rates were quite low. The poor efficiency of clavulanate as a class C inactivator appeared to rest upon a very slow acylation of the protein, and of a relatively high turnover rate. PMID- 8243631 TI - A synthetic model of collagen structure taken from bovine macrophage scavenger receptor. AB - A putative collagen structure from macrophage scavenger receptors binds to a wide range of ligands. In order to elucidate the ligand's binding mode, this collagen structure was constructed using short peptides. This was accomplished by the reaction of a tri-bromoacetylated branched peptide with a purified unprotected 25 residue peptide, which contained Cys, 4 repeats of triplet, Gly-Pro-Hyp, and 12 residues from the bovine of the N-terminus cross-linked tripeptide show that it forms a collagen structure below 10 degrees C and an extended structure at high temperature with a midpoint of 20 degrees C. PMID- 8243632 TI - Transcription in vitro promoted by the Agrobacterium VirG protein. AB - Expression of the virulence genes (vir) on the hairy-root-inducing plasmid pRiA4 is induced by plant signals in Agrobacterium cells through a two-component regulatory system, the VirA-VirG system. We constructed an in vitro transcription system that consisted of the purified VirG protein and the Agrobacterium RNA polymerase holoenzyme. Both versions of VirG, the non-phosphorylated form and the VirA-phosphorylated form, were active but showed different patterns of the pH dependency for transcriptional activation. PMID- 8243633 TI - Differential expression of dystrophin, utrophin and dystrophin-associated proteins in peripheral nerve. AB - The dystrophin-glycoprotein complex is a novel laminin receptor in skeletal muscle. Dystrophin-associated proteins are comprised of an extracellular glycoprotein of 156 kDa (156DAG), transmembrane glycoproteins of 50 kDa (50DAG), 43 kDa (43DAG) and 35 kDa (35DAG), and a cytoskeletal protein of 59 kDa (59DAP). The laminin-binding 156DAG and 43DAG are encoded by a single gene and are now called alpha- and beta-dystroglycan, respectively. In neuromuscular junctions, utrophin, an autosomal homologue of dystrophin, is associated with sarcolemmal proteins identical or immunologically homologous to the dystrophin-associated proteins. Here we demonstrate the co-localization of Dp116 (a 116 kDa protein product of the DMD gene), full-size utrophin, alpha- and beta-dystroglycan, 59DAP and 35DAG in a thin rim surrounding the outermost layer of myelin sheath of peripheral nerve fibers. The alpha-dystroglycan in peripheral nerve had molecular weight of 120 kDa instead of 156 kDa, suggesting different levels of glycosylation between skeletal muscle and peripheral nerve. In sharp contrast to skeletal muscle, however, full-size dystrophin and 50DAG were undetectable in peripheral nerve. Our results demonstrate the varied expression of the components of the dystrophin/utrophin-glycoprotein complex between skeletal muscle and peripheral nerve suggesting the complex may exist in varied compositions and have varied functions in these two tissues. PMID- 8243634 TI - Characterization of the hepatic insulin receptor undergoing internalization through clathrin-coated vesicles and endosomes. AB - Administration of insulin to rats caused a transient increase in the amount of hepatic insulin receptor present in clathrin-coated vesicles and endosomes. However, the total 'in vitro' insulin stimulated tyrosine kinase activity of the receptor present in endosomes did not vary when expressed per mg of protein and decreased when expressed per beta-subunit content. A decrease in the endogenous phospho tyrosine content of the receptor beta-subunit was observed in endosomes in response to insulin. This indicates that a fraction of the internalized receptor is dephosphorylated in endosomes, which renders it unable to become stimulated by insulin 'in vitro'. PMID- 8243635 TI - Neuron-specific glucose transporter (NSGT): CNS distribution of GLUT3 rat glucose transporter (RGT3) in rat central neurons. AB - The identity of the glucose transporters (GLUT) expressed in neurons in situ has yet to be fully established. In the present study we have isolated the GLUT3 (RGT3) cDNA and produced anti RGT3 polyclonal antibody allowing us to investigate the cellular localization and tissue distributions of RGT3 mRNA and protein in the central nervous system of the rat by the methods of in situ hybridization and immunohistochemistry. Here we demonstrate the direct evidence that RGT3 is present in neurons in adult rat brain. In situ hybridization showed the expression of RGT3 mRNA mostly in the regions of hippocampus, cerebral cortex, striatum, and the granule cell layer of the cerebellum, indicating that RGT3 mRNA is predominantly expressed within neurons. Immunohistochemistry showed that RGT3 protein is widely distributed in the rat brain, and concentrated on the plasma membrane of neurons. Double labeling studies with anti-RGT3, glial fibrillary acidic protein (GFAP), and neuron specific enolase (NSE) antibodies revealed the specific expression of RGT3 protein in neurons. Thus, RGT3 is indicated to be a neuron specific glucose transporter isoform (NSGT), and suggested to play a functionally significant role in rat central neurons. PMID- 8243636 TI - Amino acid sequence and thermostability of xylanase A from Schizophyllum commune. AB - The amino acid sequence (197 residues) of xylanase A from the fungus, Schizophyllum commune, was determined by automated analysis of peptides from proteolytic and acid cleavage. The sequence is similar to two Trichoderma xylanases (approximately 56% identical amino acids), but also shows at least 40% identities with xylanases from Bacillus subtilis, B. pumilus and B. circulans. The conserved regions of the enzyme contain only two glutamic acid residues which implicates their possible involvement in catalysis. The disulfide bond in xylanase A is not conserved in this family. In spite of this, the B. subtilis xylanase was found to be more thermostable than xylanase A. PMID- 8243637 TI - Experimental evidence for 60 Hz magnetic fields operating through the signal transduction cascade. Effects on calcium influx and c-MYC mRNA induction. AB - We tested the hypothesis that early alterations in calcium influx induced by an imposed 60 Hz magnetic field are propagated down the signal transduction (ST) cascade to alter c-MYC mRNa induction. To test this we measured both ST parameters in the same cells following 60 Hz magnetic field exposures in a specialized annular ring device (220 G (22 mT), 1.7 mV/cm maximal E(induced), 37 degrees C, 60 min). Ca2+ influx is a very early ST marker that precedes the specific induction of mRNA transcripts for the proto-oncogene c-MYC, an immediate early response gene. In three experiments influx of 45Ca2+ in the absence of mitogen was similar to that in cells treated with suboptimal levels of Con-A (1 micrograms/ml). However, calcium influx was elevated 1.5-fold when lymphocytes were exposed to Con-A plus magnetic fields; this co-stimulatory effect is consistent with previous reports from our laboratory [FEBS Lett. 301 (1992) 53 59; FEBS Lett. 271 (1990) 157-160; Ann. N.Y. Acad. Sci. 649 (1992) 74-95]. The level of c-MYC mRNA transcript copies in non-activated cells and in suboptimally activated cells was also similar, which is consistent with the above calcium influx findings. Significantly, lymphocytes exposed to the combination of magnetic fields plus suboptimal Con-A responded with an approximate 3.0-fold increase in band intensity of c-MYC mRNA transcripts. Importantly, transcripts for the housekeeping gene GAPDH were not influenced by mitogen or magnetic fields. We also observed that lymphocytes that failed to exhibit increased calcium influx in response to magnetic fields plus Con-A, also failed to exhibit an increase in total copies of c-MYC mRNA. Thus, calcium influx and c-MYC mRNA expression, which are sequentially linked via the signal transduction cascade in contrast to GAPDH, were both increased by magnetic fields. These findings support the above ST hypothesis and provide experimental evidence for a general biological framework for understanding magnetic field interactions with the cell through signal transduction. In addition, these findings indicate that magnetic fields can act as a co-stimulus at suboptimal levels of mitogen; pronounced physiological changes in lymphocytes such as calcium influx and c-MYC mRNA induction were not triggered by a weak mitogenic signal unless accompanied by a magnetic field. Magnetic fields, thus, have the ability to potentiate or amplify cell signaling. PMID- 8243638 TI - Modulation of interleukin 2 high affinity binding to human T cells by a pyrimidodiazepine insect metabolite. AB - An insect metabolite containing the little known pyrimido[4,5-b][1,4]diazepine ring system has been found to act as an effective mimic of tetrahydrobiopterin in its ability to modulate the affinity of interleukin 2 (IL-2) for its receptors on human T cells. Semi-empirical molecular orbital calculations reveal that while tetrahydrobiopterin has considerable flexibility, the pyrimidodiazepine has rather few conformational options and offers a useful model for exploring the nature of the pterin binding site. PMID- 8243639 TI - Transfection-mediated expression of human Hsp70i protects rat dorsal root ganglian neurones and glia from severe heat stress. AB - Considerable evidence suggests that the expression of heat shock proteins prior to a toxic insult (e.g. ischaemia, excitoxins, heat) can confer protection to neurones and glia. It is not certain which hsp(s) are involved in conveying these neuroprotective effects. Here we show that calcium phosphate-mediated transfection of dorsal root ganglia with an EF-1 alpha promoter-hsp70i expression vector significantly increased the survival of neurones and glia exposed to a severe heat stress. These data suggest that overexpression of hsp70i plays an important role in protecting neurones and glia from the denaturing effects of severe thermal stress. Inducing the expression of specific hsps may lead to the development of novel treatment strategies for CNS diseases. PMID- 8243640 TI - Evidence for release of free glucose from muscle during amylin-induced glycogenolysis in rats. AB - Amylin, a 37 amino acid product of pancreatic beta-cells, inhibits insulin stimulated radioglucose incorporation into muscle glycogen. In the current study, we exercised rats and then prelabelled the glycogen pool by infusing [3 3H]glucose along with glucose and insulin. Subsequent amylin administration increased the rate of appearance of tritiated water 6.4-fold, consistent with stimulation of glycogenolysis and passage of the released moieties through the hexose --> triose step of glycolysis. Further, there was an increase in plasma [3 3H]glucose after amylin, consistent with the release of free glucose previously sequestered in muscle glycogen. Calcitonin gene-related peptide (8-37), an amylin antagonist, prevented these actions. PMID- 8243641 TI - Activation of NF-kappa B by cAMP in human myeloid cells. AB - Nuclear factor kappa B (NF-kappa B) is a DNA-binding regulatory factor that controls the transcription of a number of genes. Various agents are known to activate this factor. We have studied the ability of cAMP to stimulate NF-kappa B activity in human myeloid cells. Electrophoretic mobility assay revealed that structural cAMP analogs and agents elevating intracellular cAMP levels induced NF kappa B DNA-binding activity. The inducibility was dependent on the maturation stage of myeloid cells. In promyelocytic HL-60 cells cAMP induced higher NF-kappa B activity than in more differential THP-1 cells and in human monocytes. By transfecting HL-60 and THP-1 cells with reporter constructs containing NF-kappa B DNA-binding sites, we observed that cAMP-induced NF-kappa B was transcriptionally active. PMID- 8243642 TI - Stimulation of prostaglandin production by hepatocyte growth factor in human gastric carcinoma cells. AB - Hepatocyte growth factor (HGF), a protein with pleiotropic biological activity affecting cell growth and motility, was found to markedly activate prostaglandin production in human gastric carcinoma TMK-1 cells. HPLC analysis revealed that HGF stimulated the production of prostaglandin E2 (PGE2), which is the major prostaglandin produced in these cells. HGF maximally stimulated PGE2 production at a concentration of 10 ng/ml, and it was a more potent stimulator of PGE2 production than epidermal growth factor (EGF), which is known to stimulate prostaglandin production in various cell lines. The simultaneous addition of HGF and EGF caused no further stimulation of the PGE2 production observed in HGF treated cells. We showed also that HGF increased the arachidonate release from TMK-1 cells, which release was completely suppressed by the addition of phospholipase A2 (PLA2) inhibitors. Further studies in vitro showed that HGF enhanced cellular activities of cytosolic PLA2 and cyclooxygenase 1.5-fold each. These results indicate that HGF stimulates prostaglandin production through increases in both cytosolic PLA2 and cyclooxygenase activities. PMID- 8243643 TI - The affinity-labelling of cathepsin S with peptidyl diazomethyl ketones. Comparison with the inhibition of cathepsin L and calpain. AB - Since peptidyl diazomethyl ketones are useful irreversible inhibitors for inactivating cysteinyl proteinases in vitro and in vivo and in order to reveal their role, we set out to obtain selective and effective reagents for cathepsin S. A number of such derivatives with hydrophobic amino acid residues, such as valine, leucine and tryptophane in positions adjacent to the primary specificity site were synthesized and these provided inhibitors rapidly acting at high dilution. For example, 1 nM Z-Leu-Leu-Nle-CHN2 inactivates cathepsin S with k2nd = 4.6 x 10(6) M-1 x s-1 at pH 6.5, 25 degrees C. Similarities to the specificities of cathepsin L and calpain were evident. However, Z-Val-Val-NleCHN2 is over 300 times more effective in inactivating S than L. On the other hand, Z Phe-Tyr(t-Bu)CHN2 is about 10(4) more effective against L than S. Reagents are thus now available for a clear discrimination between these proteases. PMID- 8243644 TI - Secretagogue-induced calcium wave shows higher and prolonged transients of nuclear calcium concentration in mast cells. AB - To clarify the mechanism of secretagogue (compound 48/80)-induced calcium signaling in rat peritoneal mast cells, we analyzed serial confocal calcium images with high spatial and temporal resolution using different Ca(2+)-probes. The Ca(2+)-wave began at the periphery of the cytoplasm, and then spread to the center of the nucleus. Nuclear [Ca2+]i was clearly higher than cytoplasmic [Ca2+]i. The heterogeneity of [Ca2+]i continued until about 2 min after degranulation. These results suggest the existence of an intranuclear Ca(2+) store which possesses a Ca(2+)-releasing mechanism similar to that in the cytoplasm. PMID- 8243645 TI - Antibodies to fluorylsulfonylbenzoyladenosine permit identification of protein kinases. AB - Antibodies were raised to fluorylsulfonylbenzoyladenosine (FSBA) coupled to carrier proteins. Affinity purified sera are shown to specifically detect protein kinase C-beta 1 treated with FSBA. Immunodetection is specific to modification at the ATP binding site as judged by the ability of ATP to block subsequent detection. The antisera are shown to react with a number of U937 cell proteins fractionated on Mono Q and treated with FSBA; the majority of these immunoreactions are prevented by inclusion of ATP in the FSBA treatments. One of these is positively identified as PKC-beta 1. PMID- 8243646 TI - Impaired gene transcription and nuclear protein kinase C activation in the brain and liver of aged rats. AB - The expression of the hsp70 and c-fos genes and the activation of nuclear protein kinase C (PKC) were studied in young and aged whole rats under heat-shock conditions. The induction of hsp70 and c-fos genes by heat shock were decreased several fold in the brain as well as in the liver of senescent animals. Nuclear run-off transcription assay indicated that this age-related impairment could be attributed to a block at the level of transcription. Nuclear PKC activation by heat shock was not apparent in old animals. Nuclear PKC involvement in the repression of transcription during senescence is postulated. PMID- 8243647 TI - Effects of amino acid substitution on the thermal stability of MS2 capsids lacking genomic RNA. AB - The thermal stability of capsids of the bacteriophage MS2, lacking genomic RNA, has been investigated using electron microscopy. Coat protein mutants with amino acid substitutions at residues involved in making contracts at both inter molecular interfaces and within the coat protein submit are also capable of forming 'empty' capsids of the same size and symmetry as the wild-type protein. Mutations have been characterised which are neutral, deleterious or advantageous in terms of thermal stability. In some cases, the results can be rationalised by reference to the recently refined X-ray crystal structure of the wild-type particle. PMID- 8243648 TI - Determination and mutational analysis of the phosphorylation site in the hypusine containing protein Hyp2p. AB - Electrospray mass spectrometry of the purified isoforms of the hypusine containing protein of Saccharomyces cerevisiae Hyp2p suggested a phosphorylation of the acidic isoform, which was confirmed by phosphatase treatment. The phosphorylation site was mapped to the N-acetylated serine residue in position no. 1 by mass spectrometric analysis of enzymatic fragments. Mutation of this serine residue gives rise to only the basic isoform, confirming our protein chemical data. As this mutation has no effect on cell viability or growth rate, the unphosphorylated isoform is sufficient to exert the essential in vivo function of Hyp2p. PMID- 8243649 TI - Analysis of topoisomerase II-mediated DNA cleavage of the c-myc gene during HL60 differentiation. AB - We have investigated the effect of mAMSA, a potent topoisomerase II inhibitor, on the c-myc proto-oncogene of the acute promyelocytic leukemia HL60 cell line during its differentiation. When HL60 cells were induced by dimethylsulfoxide (DMSO) to terminally differentiate, a rapid drop in the level of c-myc mRNA was observed, followed by an arrest of cell proliferation. In contrast, the level of topoisomerase II mRNA was transiently increased with a maximum at 6 h after DMSO addition and was then completely abolished after 48 h, indicating that topoisomerase II is activated during the onset of HL60 differentiation. In exponentially growing cells, treatment by mAMSA results in the formation of topoisomerase II-mediated double strand DNA breaks in the c-myc gene at positions where topoisomerase II would normally nick and reseal the two strands. In HL60 cells treated with both mAMSA and DMSO, the sites in the c-myc gene at which mAMSA had induced cleavage were not altered. However, a DNA cleavage site located at the end of the first c-myc exon (position +3100), was strongly stimulated by mAMSA and DMSO treatment. This site fell within a DNase I hypersensitive region encompassing the MYC intron factor 1 (MIF1) binding site, where transcription elongation is normally blocked during differentiation. These data indicate that a change of topoisomerase II binding to critical regulatory region of the c-myc gene is associated with the downregulation of this gene during differentiation. PMID- 8243650 TI - Nature of the pH-induced conformational changes and exposure of the C-terminal region of chromogranin A. AB - Chromogranin A is known to undergo pH induced conformational changes, and the difference in conformation is supposed to be responsible for the difference in Ca2+ binding property. To gain insight regarding the overall structure and the nature of pH-induced conformational changes of chromogranin A, limited trypsin digestions were carried out at pH 5.5 and pH 7.5. The resulting fragments were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the amino acid sequences of the tryptic fragments were determined. From these analyses it was shown that the chromogranin A structure consists of an N-terminal compact core region and a rather loosely organized C-terminal region and that the change of pH from 7.5 to 5.5 loosened the overall structure of chromogranin A, exposing the C-terminal region. Since the conserved C-terminal region (residues 407-431) was shown to exist in monomer-dimer and monomer-tetramer equilibria at pH 7.5 and 5.5, respectively, the conformational changes of the region at pH 7.5 and 5.5 were studied by circular dichroism spectroscopy using a synthetic peptide representing the conserved C-terminal region. When the pH was changed from 7.5 to 5.5, the coil structure of the C-terminal peptide decreased with an accompanying increase of alpha-helicity. PMID- 8243651 TI - Interaction between calponin and smooth muscle myosin. AB - Calponin is a thin filament-associated protein in smooth muscle that has been shown to bind actin, tropomyosin and calmodulin, and has been implicated to play a role in regulation of smooth muscle contractility. Using a centrifugation assay we found that calponin interacts with unphosphorylated filamentous smooth muscle myosin. We found that this calponin-myosin interaction is reversed by Ca(2+)-CaM, and depends on ionic strength. At 50 mM NaCl the binding constant and the stoichiometry of this interaction were estimated to be 2 x 10(6) M-1, and 1.2-2.4 calponin per myosin, respectively. We suggest that the calponin-myosin interaction could be involved in regulation of smooth muscle contractility by anchoring myosin to actin. PMID- 8243652 TI - Primary structure and functional expression of mouse pituitary and human brain corticotrophin releasing factor receptors. AB - Corticotrophin-releasing factor (CRF) is the principal hypothalamic factor governing the pituitary-adrenal axis, but the wide extra-pituitary distribution of CRF and its receptors suggest a major role for this neuropeptide in the integration of the overall physiological and behavioral responses of an organism to stress. We have cloned a CRF receptor complementary DNA (cDNA) by expression in COS-7 cells of a cDNA library from the AtT20 mouse pituitary tumour cell line. The cloned mouse cDNA was then as a probe to isolate a human CRF receptor cDNA from a human brain cDNA library. The mouse and human cDNAs both encode 415 amino acid proteins that are 97% identical, containing seven putative transmembrane domains characteristic of G protein-coupled receptors. The CRF receptor shows homology with the receptors for growth hormone-releasing factor, vasoactive intestinal peptide, secretin, parathyroid hormone, and calcitonin. COS-7 cells transfected with the mouse CRF receptor cDNA bind radiolabelled ovine CRF with high affinity and respond specifically to CRF by accumulation of intracellular cAMP. A 2.7 kb mRNA coding for the CRF receptor could be detected in AtT20 cells and human cortex tissue. PCR analysis also detected the receptor transcript in human pituitary, brainstem, and testis. PMID- 8243653 TI - Reaction of the type III iodothyronine deiodinase with the affinity label N bromoacetyl-triiodothyronine. AB - The type III iodothyronine deiodinase (ID-III) catalyzes the inner ring deiodination and, thus, the inactivation of the thyroid hormones T4 and T3. ID III activity in rat brain, rat placenta and embryonic chicken liver is inhibited by the affinity label N-bromoacetyl-T3 (BrAcT3) with an affinity similar to that of T3. Reaction of rat brain and placenta microsomes with BrAc[125I]T3 resulted in the extensive labeling of a 32 kDa protein (p32). However, p32 was also prominently labeled in fetal rat liver microsomes which have no ID-III activity. Labeling of p32 was not influenced by 100 microM substrate analogs or inhibitors of ID-III, some of which completely inhibit ID-III activity at 1 microM. BrAc[125I]T3 labeling of embryonic chicken liver microsomes did not reveal p32 or another protein possibly related to ID-III. In contrast to previous suggestions, it is unlikely that p32 represents ID-III or a subunit thereof. PMID- 8243654 TI - Neurogranin, a B-50/GAP-43-immunoreactive C-kinase substrate (BICKS), is ADP ribosylated. AB - Neurogranin is a neurone-specific, B-50-immunoreactive C-kinase substrate that has limited homology to, but considerable biochemical similarity to B-50/GAP43. The most significant differences between these two proteins are their cellular localisation and molecular mass (Neurogranin, 7.5 kDA cytosolic; and B-50, 25 kDa membranal). An understanding of the similarities and differences between Neurogranin and B-50 may facilitate the elucidation of their hitherto elusive functions in the nervous system. The results of the present study demonstrate that, in common with B-50, Neurogranin is a substrate for ADP-ribosyltransferase. This finding is discussed with regard to the concept of molecular flexibility of B-50-like proteins as the basis of their putatively diverse roles in the nervous system. PMID- 8243655 TI - Lymphotoxin gene expression by melanocytes and melanoma cell lines and persistence of unspliced mRNA. AB - Human melanoma cell lines express many different cytokines [1], including lymphotoxin (LT), the production of which has been considered to be restricted to cells of the lymphocytic lineage in response to cell activation. LT expression by melanomas is constitutive and characterized by the presence of two mRNAs. In the present paper we report an analysis of the origin of the two LT-specific transcripts in four human melanoma cell lines at different stages of progression and in four melanocytic cell lines. Reverse transcription-polymerase chain reaction (RT-PCR) performed with primers lying in the first and fourth exons and hybridization with intron probes showed a spliced and a full-unspliced LT mRNA. This pattern was also displayed by one of four melanocyte cell lines. Western blot analysis indicated that LT RNA is properly translated to a 23-25 kDa protein and immunocytochemistry showed its localization within the cytoplasm and on the cell membrane. PMID- 8243656 TI - Variant plasma gelsolin responsible for familial amyloidosis (Finnish type) has defective actin severing activity. AB - Familial amyloidosis, Finnish type is caused by a single base mutation in gelsolin, an actin filament severing and capping protein that is present in most tissues and in blood plasma. The mutation replaces aspartic acid with asparagine at residue 187 of the plasma sequence. This renders the gelsolin susceptible to proteolysis as a consequence of which amyloid protein is formed. Here it is shown that the mutant protein in plasma from a patient homozygous for this mutation lacks both actin severing and nucleating activities. Evidence is presented that the cleaved mutant gelsolin has dissociated under non-denaturing conditions and that the resultant 65,000 and 55,000 M(r) C-terminal fragments aggregate. PMID- 8243657 TI - Cytochrome d axial ligand of the bd-type terminal quinol oxidase from Escherichia coli. AB - Using various spectroscopic techniques, we studied the structure of the dioxygen reduction site of the bd-type terminal quinol oxidase in the aerobic respiratory chain of Escherichia coli. Resonance Raman and FT-IR spectroscopies identified the v(Fe(2+)-CO) and v(C-O) stretching frequencies at 471 and 1980.7 cm-1, respectively, at the cytochrome d center of the dithionite-reduced CO-bound enzyme. The CO ligation in the cytochrome bd complex is considerably different from those of the heme-copper terminal oxidases. Anaerobic addition of NO to the air-oxidized enzyme caused an exchange of cytochrome d-bound dioxygen with NO leading to an appearance of cytochrome d-NO EPR signal. But there is no superhyperfine structure originating from the cytochrome d proximal 14N ligand in the central resonance of the NO EPR signal. These results suggest that cytochrome d axial ligand of the cytochrome bd complex is likely a histidine residue in an anomalous condition or other than a histidine residue and, therefore, the molecular structure around the dioxygen-binding site is different from that of the heme-copper terminal oxidases. PMID- 8243658 TI - Glycoprotein IIb peptide 656-667 mimics the fibrinogen gamma chain 402-411 binding site on platelet integrin GPIIb/IIIa. AB - The human integrin glycoprotein IIb/IIIa complex plays a central role in haemostasis as an inducible receptor for fibrinogen and other adhesive proteins at the platelet plasma membrane. Current evidence indicates that the ligand binding domain of GPIIb/IIIa is discontinuous and placed at the subunit interface. Here we show that a synthetic peptide containing the polypeptide stretch GPIIb 656-667, which is hidden within the resting platelet GPIIb/IIIa heterodimer but becomes exposed following platelet activation with thrombin, binds to soluble fibrinogen (n = 2.3 +/- 1.3; Kd = 2 +/- 0.8 x 10(-5) M). This interaction is Ca(2+)-independent and can be partially inhibited with synthetic fibrinogen gamma-chain peptide 400-411 but not with GRGDS. In addition, peptide GPIIb 656-667 inhibits in a dose-dependent manner the aggregation of activated platelets (IC50 = 170 microM). Altogether, our results indicate that the GPIIb 656-667 region may form part of the inducible fibrinogen binding site and may not overlap with the integrin RGD-recognition domain. PMID- 8243659 TI - Comment on the paper by Shchelkunov et al. (1993) FEBS Letters 319, 80-83. Two genes encoding poxvirus cytokine receptors are disrupted or deleted in variola virus. PMID- 8243660 TI - NMR determination of the secondary structure and the three-dimensional polypeptide backbone fold of the human sterol carrier protein 2. AB - Nuclear magnetic resonance (NMR) spectroscopy was used to determine the secondary structure and the three-dimensional polypeptide backbone fold of the human sterol carrier protein 2 (hSCP2), which is a basic protein with 123 residues believed to participate in the intracellular transport of cholesterol and various other lipids. Sequence-specific assignments were obtained for nearly all backbone 1H and 15N resonances, as well as for about two-thirds of the side-chain 1H resonances, using uniform 15N-labeling of the protein combined with homonuclear two-dimensional 1H NMR and three-dimensional 15N-correlated 1H NMR. Three alpha helices comprising the polypeptide segments of residues 9-22, 25-30 and 78-84 were identified by sequential and medium-range nuclear Overhauser effects (NOE). The analysis of long-range backbone-backbone NOEs showed that hSCP2 further contains a five-stranded beta-sheet including the residues 33-41, 47-54, 60-62, 71-76 and 100-102, which is a central feature of the molecular architecture. The first three strands are arranged in an antiparallel fashion, the polypeptide chain then crosses over this three-stranded sheet in a right-handed sense so that the fourth strand is added parallel to the first one. The fifth strand runs antiparallel to the fourth one, so that the overall topology is +1, +1, -3x, -1. The three-dimensional arrangement of the beta-sheet and the first two helices was determined using an input of 625 NOE upper distance constraints and 95 scalar coupling constants for a preliminary structure calculation with the distance geometry program DIANA. PMID- 8243661 TI - Immediate-early gene induction by repetitive mechanical but not electrical activity in adult rat cardiomyocytes. AB - Mechanical factors are thought to play an important role in the induction of myocardial hypertrophy. Yet, it is not known whether active contraction induces genes that probably represent initial steps in the hypertrophic response in the adult myocardium--and if so, whether the mechanical or the electrical component of the twitch governs this response. We therefore investigated whether electrical stimulation of contraction was able to induce the immediate-early genes (IEGs) egr-1 and c-fos in adult rat cardiomyocytes. Cyclical contraction led to an increase in egr-1 and c-fos mRNA levels within 30 min. Full inhibition of contraction during electrostimulation by the Ca(2+)-desensitizer 2,3-butanedione monoxime (BDM) totally blocked this IEG-response without altering membrane potential. These data suggest that in adult myocardium, the mechanical rather than the electrical activity is responsible for the IEG-response during active twitch. PMID- 8243662 TI - The MyoD binding site is dispensable for cardiac actin gene expression in the somites of later stage Xenopus embryos. AB - We have studied the transcription directed by the promoter of a Xenopus borealis cardiac actin gene fused to a globin reporter sequence after injection into X. laevis embryos. This promoter is known to be effective specifically in somites of neurula stage embryos, and work on a very similar X. laevis gene has indicated the importance of an upstream MyoD binding site (E box) in this process. We show that, although in the absence of the MyoD site there is a small amount of tissue specific expression in neurulae, transcription is abundant in the tailbud embryo a few hours later. Thus, although the E box is initially essential, other sequences must adopt the same role soon afterwards. PMID- 8243663 TI - Adaptability of nonnatural aromatic amino acids to the active center of the E. coli ribosomal A site. AB - 3'-N-Aminoacyl analogs of puromycin with nonnatural aromatic amino acids were synthesized and their inhibitory activity in E. coli in vitro protein synthesizing system was evaluated. The analogs with L-2-naphthylalanine, L-p biphenylalanine, L-2-anthrylalanine and trans-L-p-phenylazophenylalanine were found to inhibit the protein synthesis with high efficiency. The inhibition suggests that these nonnatural amino acids are accepted by the active center of the E. coli ribosomal A site. A model for the adaptability of nonnatural aromatic amino acids to the active center is proposed. PMID- 8243664 TI - Alternative transcripts of the human CD23/Fc epsilon RII. A possible novel mechanism of generating a soluble isoform in the type-II cell surface receptor. AB - Human CD23/Fc epsilon RII is a 45 kDa type-II membrane glycoprotein having two isoforms (a and b) that only differ in the structures of their intracytoplasmic tails. CD23/Fc epsilon RII has been demonstrated to have multiple roles in the immune system such as regulation of lymphocyte growth and differentiation and IgE mediated immune responses. Here, we found that the human B-cell line RPMI8866, in addition to a and b transcripts, contained shorter transcripts (a' and b') that lack the entire third exon. These alternative transcripts were also detected in peripheral blood lymphocytes as well as other hematopoietic cell lines with CD23/Fc epsilon RII. Because exon 3 encodes all of the transmembrane segment and the anchoring region of the cytoplasmic tail, it is suggested that a' and b' transcripts encode secretory forms of CD23/Fc epsilon RII or they may function as regulatory transcripts involved in the control of CD23/Fc epsilon RII expression. PMID- 8243665 TI - The Escherichia coli cysG gene encodes the multifunctional protein, siroheme synthase. AB - Previously, the E. coli cysG gene product had been shown to sequentially methylate uro'gen III to produce precorrin-2, hence it was given the trivial name uro'gen III methylase. We now report that in addition to methylase activity, the CysG protein catalyses both the NAD+ dependent oxidation of precorrin-2 to sirohydrochlorin, but also the insertion of iron into this oxidized intermediate, thereby producing siroheme. Thus CysG is a multifunctional protein solely responsible for siroheme synthesis from uro'gen III in E. coli, and accordingly is renamed siroheme synthase. PMID- 8243666 TI - The genomic region encoding toxin gamma from the scorpion Tityus serrulatus contains an intron. AB - The gene encoding toxin gamma from the scorpion, Tityus serrulatus, was amplified by PCR from genomic DNA employing synthetic oligonucleotides designed from the reported cDNA sequence. The nucleotide sequence of this gene reveals the presence of an intron of 475 base pairs (bp) which interrupts the region that encodes the signal peptide of the precursor toxin. A comparison of the intron boundary sequences of the gamma toxin gene with ones from other arachnid genes is also presented. PMID- 8243667 TI - Synthesis of functional GABAA receptors in stable insect cell lines. AB - We have synthesised the beta 1-subunit of the bovine GABAA receptor in stable, continuous insect (Spodoptera frugiperda) cell lines. A cDNA was integrated randomly into the insect cell genome under control of a baculovirus immediate early (IE-1) gene promoter. Transformed cells were obtained by co-transfection of the insect cells with pIEK1.GR beta 1, encoding the beta 1 subunit cDNA, and pIEK1.neo, encoding the neomycin resistance gene. G-418-resistant clones were selected and expanded into continuous cell lines synthesising functional, GABA gated, homo-oligomeric chloride channels. These cell lines had significant advantages over the transient baculovirus expression system for the characterisation of receptors using electrophysiological recording techniques. PMID- 8243668 TI - Variability patterns of dehydrogenases versus peptide hormones and proteases/antiproteases. AB - Comparisons of human/rat species variants of 23 dehydrogenases versus 41 entries for peptide hormones and 41 for proteases/antiproteases reveal characteristic patterns. Dehydrogenases are 'constant' (2-8% residue differences between homologues) or 'variable' (12-40% differences). Peptide hormones exhibit a wider range, with many 'strictly conserved' and others spreading upscale, while proteases/antiproteases are overall shifted upscale, with few 'constant' and many 'variable'. Overall, the influence of polypeptide size and function is confirmed, while for the dehydrogenases, the data highlight the 'constant' pattern as the one with high frequency values and suggest for classic liver alcohol dehydrogenase the 'variable' pattern to correlate with emerging functions. PMID- 8243669 TI - RPII15 codes for the M(r) 15,000 subunit 9 of Drosophila melanogaster RNA polymerase II. AB - The RPII15 gene product of Drosophila melanogaster, which has recently been identified by sequence comparison, possesses a high similarity to subunit 9 of yeast RNA polymerase II. Using the polymerase chain reaction the coding region of RPII15 was isolated from genomic DNA of adult flies. Sequence analysis shows four amino acid substitutions in comparison to the previously reported sequence. Antisera were generated against bacterially expressed RPII15 and were used for immunoblotting experiments with RNA polymerase II of Drosophila melanogaster. This analysis identified the M(r) 15,000 subunit 9 as gene product of RPII15. PMID- 8243670 TI - Activation of snake venom metalloproteinases by a cysteine switch-like mechanism. AB - The cDNAs of several snake venom zinc endopeptidases code for a putative propeptide, which includes the conserved cysteine-containing sequence PKMCGVT. It has been suggested that binding of the cysteine thiol function to the active-site zinc, resulting in inactivation of the catalytic domain, occurs in a mode similar to the 'cysteine switch' mechanism proposed for matrix metalloproteinases. In order to confirm this hypothesis, inhibition kinetics have been performed on the metalloproteinase adamalysin II of the venom of the snake Crotalus adamanteus using several cysteine peptides. Among these the synthetic hexapeptide PKMCGV NH2, corresponding to the conserved sequence portion of the known propeptides, was found to be by far the strongest inhibitor of this proteinase with a Ki of 3.4 microM. The inhibitory potencies of an equivalent peptide with the L-Cys replaced by a D-Cys or by an L-Ser as well as of reduced glutathione, cysteine and two unrelated cysteine peptides were by one to two orders of magnitudes lower. These findings strongly support a cysteine switch-like mechanism even for activation of the snake venom metalloproteinases. PMID- 8243671 TI - Enzymes of the cyclic GMP metabolism in bovine retina. I. Cloning and expression of the gene for guanylate kinase. AB - Guanylate kinase (EC 2.7.4.8) catalyzing the reaction GMP + ATP = GDP + ADP, was purified to homogeneity from bovine retina. Using oligonucleotides based on the amino acid sequence of this enzyme, the cDNA encoding guanylate kinase (GK) was isolated and its nucleotide sequence was determined. Expression of the GK cDNA in E. coli, and the purification and functional characterization of the expressed enzyme are presented. It is shown that bovine retinal GK, like its yeast counterpart, contains the characteristic glycine-rich motif and all the amino acids involved in GMP binding. Bovine retinal enzyme is extended for several amino acid residues both at the N- and C-termini, compared to the yeast enzyme. PMID- 8243672 TI - Subunit structure of ATP synthase from Chloroflexus aurantiacus. AB - An ATP synthase has been isolated from green nonsulfur photosynthetic bacterium Chloroflexus aurantiacus, a representative of a lower branch of eubacteria. The enzyme, reconstituted with the bacterial lipids into proteoliposomes, is shown to catalyze [32P]Pi-ATP exchange (at a rate of 180 nmol [32P]ATP/min/mg). The ATP synthase is composed of nine polypeptide species (60, 50, 33, 19, 16.5, 15.5, 14.5, 13, and 8 kDa as determined by urea-SDS-PAGE). The catalytic part of the ATP synthase (which is detached by chloroform treatment) contains the first four polypeptides. In the intact ATP synthase the 14.5 and 13 kDa polypeptides are connected by disulfide bonds to form a heterodimer of 25 kDa. PMID- 8243673 TI - Generation of beta A4 from the amyloid protein precursor and fragments thereof. AB - The cellular mechanisms underlying the generation of beta A4 in Alzheimer's disease and its relationship to the normal metabolism of the amyloid protein precursor (APP) are unknown. In this report, we show that expression of the C terminal 100 residues of APP, with (SPA4CT) or without (A4CT) a signal sequence in the N-terminal position, in human neuroblastoma cells results in secretion of a 4 kDa beta A4-like peptide. In A4CT and SPA4CT expressing SY5Y cells, beta A4 generation could not be inhibited by the lysosomotropic amines chloroquine and ammonium chloride but was inhibited by brefeldin A, monensin and methylamine. The last also selectively inhibits APP secretion in neuroblastoma cells [1]. The finding that chloroquine and ammonium chloride inhibit beta A4 generation from full length APP but not from A4CT and SPA4CT are consistent with the assumption that the two cleavages necessary to generate beta A4 operate in two different compartments. Our data suggest the cleavage which generates the C-terminus of beta A4 takes place in the same compartment (late Golgi or endosomal vesicles) in which the APP-secretase operates. PMID- 8243674 TI - Carbohydrate isoforms of antithrombin variant N135Q with different heparin affinities. AB - We have changed one of the carbohydrate-bearing asparagine residues of human antithrombin to glutamine by site-directed mutagenesis and expressed the variant antithrombin, N135Q, in baby hamster kidney cells. Two isoforms were secreted, both of which had higher affinity for heparin than human plasma alpha antithrombin. Both forms had normal inhibitory activity toward factor Xa and showed normal heparin acceleration of proteinase inhibition. The mutation resulted in a higher production of the very high affinity form from about 30% to 60% of the total secreted antithrombin. This form should be the most useful for comparison of the effects of other mutations on heparin binding and proteinase inhibition. PMID- 8243675 TI - Amino acid sequence surrounding the retinal-binding site in retinochrome of the squid, Todarodes pacificus. AB - Squid (Todarodes pacificus) retinochrome was reduced to N-retinyl protein with borane dimethylamine and cleaved by CNBr. The retinyl peptide was then isolated by chromatography while being monitored for absorbances at 215 and 330 nm, and the N-terminal amino acid sequence was determined to be Ser-Lys-Thr-Gly-X-Ala-Leu Phe-Pro. This sequence was the same that we had observed at the 7th transmembrane domain of retinochrome whose structure was reported previously. During Edman degradation of the retinyl peptide, the yield of the PTH-lysine at the second cycle was lower than those of the other PTH-amino acids, proving that the lysine residue forms a Schiff's base with retinal (Lys-275 in retinochrome). The amino acid sequence surrounding the retinal-binding lysine in retinochrome greatly differed from those in a variety of known visual pigments. This fact would be associated with the difference in the photoisomerization of chromophore between retinochrome and rhodopsin. The protein structure of retinochrome is also compared with that of rhodopsin in Todarodes. PMID- 8243676 TI - Botulinum neurotoxins serotypes A and E cleave SNAP-25 at distinct COOH-terminal peptide bonds. AB - SNAP-25, a membrane-associated protein of the nerve terminal, is specifically cleaved by botulinum neurotoxins serotypes A and E, which cause human and animal botulism by blocking neurotransmitter release at the neuromuscular junction. Here we show that these two metallo-endopeptidase toxins cleave SNAP-25 at two distinct carboxyl-terminal sites. Serotype A catalyses the hydrolysis of the Gln197-Arg198 peptide bond, while serotype E cleaves the Arg180-Ile181 peptide lineage. These results indicate that the carboxyl-terminal region of SNAP-25 plays a crucial role in the multi-protein complex that mediates vesicle docking and fusion at the nerve terminal. PMID- 8243677 TI - The association of antithyroid antibodies in euthyroid nonpregnant women with recurrent first trimester abortions in the next pregnancy. AB - OBJECTIVE: To evaluate the prognostic value of antithyroid antibodies in euthyroid women with a history of recurrent first trimester abortions on future pregnancy loss. DESIGN: The sera of 42 euthyroid women with a history of three or more consecutive first trimester abortions were evaluated for the presence of antibodies to thyroglobulin and thyroid peroxidase before pregnancy and again as soon as the diagnosis of pregnancy was made. SETTING: Medical school-affiliated private infertility center. PATIENTS: Forty-two women with a history of three or more consecutive first trimester abortions who were planning to conceive again. MAIN OUTCOME MEASURE: The presence of antithyroid antibodies in the nonpregnant state and their association with pregnancy loss in the next gestation. RESULTS: Thirteen of 42 women (31%) were positive for the presence of antithyroid antibodies at the initial screening before pregnancy. All 13 maintained positivity by the time their next pregnancy was diagnosed. Only 12 of those 42 women (29%) experienced a first trimester abortion. Eight of these 12 women (67%) were positive for one or more antithyroid antibody. In contrast, among 30 nonaborting women, only 5 of 30 (17%) exhibited thyroid antibody positivity. The detection of thyroid antibodies before conception carried an increased risk of pregnancy loss in the next pregnancy (8 of 13, 62% versus 4 of 29, 14%). CONCLUSION: The presence of antithyroid antibodies in nonpregnant women with a history of recurrent abortion identifies a subgroup of women at significantly increased risk for yet another pregnancy loss in their next gestation. Because organ-specific autoantibodies thus demonstrate similar prognostic significance to nonorgan-specific autoantibodies, it is tempting to conclude that peripheral autoantibody abnormalities seen in habitual aborters only reflect an underlying T lymphocyte defect, which may be the actual cause of pregnancy loss. PMID- 8243678 TI - Study on the presence of homocysteine in ovarian follicular fluid. AB - OBJECTIVE: To study the presence of homocysteine, methionine and the vitamins folate, B12, and B6 in human ovarian follicular fluid (FF). DESIGN: Measurement of homocysteine, methionine, folate, and vitamins B12 and B6 in ovarian FF and blood. SETTING: Academic Department of Obstetrics and Gynecology at St. Radboud Hospital, Nijmegen, The Netherlands. PARTICIPANTS: Fourteen healthy women undergoing an IVF program. RESULTS: Detectable amounts of homocysteine and methionine were found in FF. Homocysteine concentrations were similar to those in serum. Methionine concentrations proved to be slightly but significantly lower than in corresponding serum samples. Concentrations of vitamins B12 and B6 were significantly lower in FF than in serum, whereas folate concentrations were not significantly different. A statistically significant correlation between corresponding serum and FF concentrations of homocysteine, folate, and vitamin B12 could be established. CONCLUSIONS: These data support the hypothesis that the ovum might be exposed to high homocysteine or low methionine concentrations, or both, and a lack of vitamins, which might be important in fertilization and early embryogenesis. PMID- 8243679 TI - Effects of the repetitive administration of progesterone by nasal spray in postmenopausal women. AB - OBJECTIVE: To study the effects of 10 days of nasal spray P treatment on P serum levels and the endometrium. DESIGN: Prospective. SETTING: University Medical School. PATIENTS: Eight postmenopausal women received oral conjugated estrogens at a daily dose of 0.625 mg for 4 weeks immediately before vaginal surgery for prolapse. For the first 9 of the last 10 days the patients also received a nasal spray dosage of 11.2 mg P three times a day; on the 10th day they received only one dose. MAIN OUTCOME MEASURES: Blood samples were taken at 8:00 A.M. on treatment days 1, 3, 5, 7, 10, and 11 to follow P serum concentration levels. Endometrial samples for histologic examination were collected before P administration and immediately after surgery to evaluate the end-organ effect. RESULTS: Mean P serum levels increased sixfold after 9 days of nasal spray P administration [from 0.612 +/- 0.280 ng/mL (1.958 +/- 0.896 nmol/L) to 3.925 +/- 1.553 ng/mL (12.560 +/- 4.970 nmol/L)] and declined thereafter, returning to the before treatment levels 24 hours after the last administration. In all subjects, the first histologic evaluation showed proliferative endometrium; the second showed clear secretive changes. CONCLUSIONS: Repetitive nasal spray P administration for 10 days in postmenopausal women led to increasing P serum levels and, when the estrogen stimulation was adequate, to secretory changes in the endometrium (end-organ effect). PMID- 8243680 TI - Functional differentiation in progesterone secretion by granulosa versus cumulus cells in the human preovulatory follicle and the effect of different induction of ovulation protocols. AB - OBJECTIVE: To investigate P secretion by granulosa cells (GCs) versus cumulus cells derived from human preovulatory follicles. DESIGN: Cells were recovered by aspiration of preovulatory follicles in 44 women participating in an IVF program. Induction of ovulation was performed using clomiphene citrate, hMG and hCG (group I), hMG/hCG (group II), buserelin acetate/hMG/hCG (group III), or Decapeptyl/hMG/hCG (group IV). SETTING: Laboratories of the IVF Unit at the Department of Obstetrics and Gynecology, Carmel Medical Center, Haifa, Israel. MAIN OUTCOME MEASURES: Secretion of P was examined after cultures for 96 hours under nonstimulated and hCG stimulated conditions. RESULTS: Progesterone secretion by GCs derived from all four groups was found to be higher compared with the respective cumulus cells. However, although the ratios of P secretion by GCs versus cumulus cells in groups I, II, and III were very similar, a significantly lower value was observed in group IV. The response of GCs to hCG in terms of P secretion was higher with at least one dose of hCG in groups I and IV compared with groups II and III. The response of cumulus cells to hCG was absent regardless of the treatment protocol used in vivo. CONCLUSION: Our results demonstrate that in the human preovulatory follicle, GCs and cumulus cells differ in their capacity to secrete P as well as in their response to hCG. They further suggest that the mode of induction of ovulation affects the relative capacity of GCs and cumulus cells to secrete P and their ability to respond to hCG. PMID- 8243681 TI - The effect of fertility drugs and in vitro methods on the outcome of 106 triplet pregnancies. AB - OBJECTIVE: To compare the effect of fertility drugs and IVF on the outcome of triplet pregnancies. DESIGN: Prospective clinical study. SETTING: A single university medical center. PATIENTS: One hundred six consecutive triplet pregnancies treated from 1984 through 1992. MAIN OUTCOME MEASURES: The frequency of pregnancy loss, livebirths, and antenatal and neonatal complications was compared in spontaneous, clomiphene citrate (CC), menotropins, and IVF triplet pregnancies. RESULTS: Eighty-one of the 106 (76.4%) triplet pregnancies progressed beyond 25 weeks, comprising 6 of 7 (85.7%) spontaneous pregnancies, 13 of 16 (81.2%) CC induced, 44 of 56 (78.6%) menotropin induced, and 18 of 27 (66.6%) IVF gestations. There were no significant differences in the stillbirth and neonatal mortality rates according to the mode of conception. The mean gestational ages and the mean birth weights were similar in the four groups. The frequency of premature contractions, premature rupture of membranes, cesarean section, and neonatal complications were similar in the ovulation induction and IVF pregnancies. CONCLUSION: Triplet pregnancies after ovulation induction and IVF have a similar outcome. PMID- 8243682 TI - Transrectal ultrasonography of infertile men. AB - OBJECTIVE: To determine normal transrectal ultrasonographic anatomy in young men and the frequency of abnormalities in the infertile population. DESIGN: Transrectal ultrasonography was performed upon 30 fertile volunteers and 150 consecutive men referred for male factor infertility. SETTING: A male fertility center. RESULTS: Transrectal ultrasonography was normal in 60% of controls and 53% of infertile group. The frequency of hyperechoic lesions within the prostate was similar in controls (40%) and infertile men (39%). Mullerian duct cysts were present in 11% of the infertile men and none of the volunteers. Rectal exam was normal in all of the men. CONCLUSIONS: Transrectal ultrasonography is more sensitive at detecting abnormalities of the seminal vesicles and prostate than rectal exam. Hyperechoic lesions within the prostate is frequently a normal finding. Mullerian duct cysts are more frequently observed in infertile men and may be a cause of ejaculatory duct obstruction. PMID- 8243683 TI - Long-term follow-up of couples after hamster egg penetration testing. AB - OBJECTIVE: To determine the clinical usefulness of the zona-free hamster egg penetration test as a long-term prognostic indicator for future pregnancy. SETTING: Division of Reproductive Endocrinology and Infertility at the Washington University Medical Center. PARTICIPANTS: All couples (n = 148) who had a hamster egg penetration assay performed between March 1, 1985 and December 31, 1986 were identified and followed with direct or telephone contact up to 68 months after the initial assay. MAIN OUTCOME MEASURE: The monthly fecundity rates using life table analysis and the 5-year incidence of pregnancy were categorized by the percentage of hamster eggs penetrated and by history of previous urologic surgery. RESULTS: There were no significant differences in the rate nor incidence of pregnancy in couples with hamster egg penetration scores of 0%, > 0% and < or = 10%, > 10% and < or = 20%, or > 20%. Although men with previous urologic surgery tended to have lower scores, there was no significant difference in the 5 year incidence of pregnancy. CONCLUSION: The hamster egg penetration score is not predictive of incidence of pregnancy nor time to conception. PMID- 8243684 TI - Detection of X- and Y-bearing human spermatozoa after motile sperm isolation by swim-up. AB - OBJECTIVE: To assess the ratio of X- to Y-bearing human spermatozoa in motile fractions isolated by the swim-up technique. DESIGN: The proportions of X- and Y bearing sperm were determined in neat semen samples (control) and in motile fractions isolated from the same samples by swim-up. X- and Y-bearing sperm were simultaneously identified using chromosome-specific DNA probes and double fluorescence in situ hybridization. SETTING: Hospital-based university department. PARTICIPANTS: Ten healthy donors with normal semen characteristics. MAIN OUTCOME MEASURES: The distribution of haploid cells (X or Y), normal size cells with two sex chromosome (XX, YY, or XY), and large cells containing two (XX, YY, or XY) or four (XXYY) sex chromosomes were measured in neat semen samples and in motile fractions prepared by swim-up. RESULTS: Overall, 95% of sperm in the neat semen and swim-up fractions were labeled with the probes. The ratios of X- to Y-bearing sperm were 47.3:46.9 (neat semen) and 48.4:47.1 (swim up fractions), which were not significantly different from a 1:1 ratio. The frequencies of sperm with normal size nuclei and two sex chromosomes (XX, YY, or XY) in the swim-up fractions were not significantly different from the controls, but there was a significant reduction in the proportion of cells with large nuclei and two (XX, YY, or XY) or four (XXYY) sex chromosomes in the swim-up fractions. CONCLUSIONS: The swim-up technique does not selectively enrich either X- or Y-bearing sperm. Because the isolation of motile spermatozoa is an important procedure for routine IUI, IVF-ET, and GIFT, the results of this study are important reassurance that the sex ratio is not altered by this method of sperm preparation. PMID- 8243685 TI - Efficacy of kallikrein in the treatment of oligozoospermia and asthenozoospermia: a double-blind trial. AB - OBJECTIVE: To assess the effect of kallikrein on sperm motility and sperm count in infertile oligozoospermic and asthenozoospermic men. DESIGN: Double-blind placebo-controlled clinical trial. SETTING: Multicenter trial in three andrologic clinics. PATIENTS: Infertile men with < 20 x 10(6) total sperm count and > 3 x 10(6) sperm cells/mL and/or sperm motility of < 30%. INTERVENTIONS: One hundred fourteen men received either placebo or 600 kallikrein units of kallidinogenase by mouth daily for 12 weeks. Semen analyses were performed before and at 6-week intervals during the trial. Compliance and incidence of adverse effects were assessed. RESULTS: The incidence of adverse effects was similar under placebo and under kallidinogenase treatment, usually mild, transient, and did not require the cessation of treatment in any patient. Kallidinogenase was not superior to placebo in improving sperm count and sperm motility. CONCLUSIONS: At the doses and for the duration given, our results were disappointing. The absence of a therapeutical effect, as evidenced in the present study may be due to the very low absorption rate of kallidinogenase after oral administration. Probably a much higher dose of kallidinogenase should be used to achieve a therapeutic affect. PMID- 8243686 TI - Subzonal sperm insemination and total or extreme asthenozoospermia: an effective technique for an uncommon cause of male infertility. AB - OBJECTIVE: To assess the value of subzonal insemination (SUZI) in cases of total or extreme asthenozoospermia. DESIGN: The results of IVF after SUZI were assessed in six cycles (four couples) with documented asthenozoospermia. SETTING: Institut de Medecine de la Reproduction, Marseille, France. PATIENTS: Four couples underwent six retrieval cycles. In all cases asthenozoospermia was total (100% of immotile spermatozoa) or extreme (5% nonprogressive motility). Oligozoospermia and teratozoospermia were ruled out in all cases. In two cases electron microscopy revealed the absence of the central singlet in the flagellum axonemal complex and in one case a major dysplasia of the fibrous sheath in the flagellum principal piece. RESULTS: The overall fertilization rate was 45%. The diploid embryo rate was 33%. Embryo quality was satisfactory. In five of six cycles, three embryos were transferred. Two triplet pregnancies were obtained corresponding to an implantation rate per transferred embryo of 40%. CONCLUSION: Total or extreme asthenozoospermia seems to be a good indication for SUZI. PMID- 8243687 TI - Comparison of two androgens plus depot-medroxyprogesterone acetate for suppression to azoospermia in indonesian men. World Health Organization. Task Force on Methods for the Regulation of Male Fertility. AB - OBJECTIVE: To compare the sperm suppression in Indonesian men achieved by combination treatment with either T enanthate or 19-nortestosterone ester) with depot-medroxyprogesterone acetate (depot-MPA). DESIGN: Prospective multicenter parallel group randomized trial. SETTING: Five centers in Faculties of Medicine in Indonesia. PARTICIPANTS: Ninety-six normal healthy volunteers. INTERVENTIONS: Seven weekly injections of T enanthate or 19-nortestosterone ester, 200 mg IM, followed by injections every 3 weeks to week 24. In both groups 250 mg depot-MPA was injected IM at 0, 6, 12, and 18 weeks. MAIN OUTCOME MEASURES: Sperm suppression by semen analysis at 3-week intervals and clinical chemistry, hematology, and gonadotrophin hormones monitored at 6-week intervals. Recovery assessed monthly until return to normal sperm concentration (> or = 20 x 10(6)/mL). RESULTS: Eighty-seven of 90 men (96.7%) achieved consistent azoospermia; 95.6% (43/45) and 97.8% (44/45) in the T and 19-nortestosterone groups, respectively. The remaining three men also suppressed to near azoospermic levels. Six of 96 men discontinued injections for failure to attend at the schedule time (3) and for medical reasons (3). Plasma FSH and LH concentrations were suppressed during treatment and returned to baseline levels during the recovery period. Clinical chemistry and hematologic values remained in the normal ranges throughout. More men complained of loss of libido in the 19 nortestosterone group (seven versus two, respectively). Median time to recover to normal sperm concentrations was 6.5 months. CONCLUSIONS: The two treatments were equally effective in sperm suppression. The rates of azoospermia achieved were higher than in previous studies with non-Indonesian men. PMID- 8243688 TI - Prospective study of leukocytes and leukocyte subpopulations in semen suggests they are not a cause of male infertility. AB - OBJECTIVE: To determine the effects of leukocytes in semen on sperm quality and the ability to achieve conception. DESIGN: A prospective analysis of 512 couples attending a regional infertility clinic. Leukocyte subsets were quantified using a monoclonal antibody-based staining procedure. In addition to basic seminal parameters (density, motility, morphology, and antisperm antibodies), reactive oxygen species and immature germ cells were also quantified in the semen of each patient. The presence or absence of a treatment-independent conception was determined 22 months after the start of the study. Semen parameters were then related to the ability to conceive. SETTING: University-based center for reproductive medicine. PARTICIPANTS: Success or failure to conceive was recorded from 512 couples. Couples were then selected to minimize the influence of any pathology of the female on outcome. A final study group of 229 couples, in which the women had regular menstrual cycles, ovulatory midluteal serum P levels of > 18 nmol/L, and patent fallopian tubes was finally selected for analysis. MAIN OUTCOME MEASURE: Pregnancy. RESULTS: Leukocyte concentration (total or individual subsets) was not associated with either reduced semen quality or conception rates. Similarly, neither reactive oxygen species or antisperm antibody (immunobead) concentration had any bearing on the outcome. Of all semen parameters measured, only the level of immature germ cells was found to be negatively associated with the rate of conception. CONCLUSION: Measurement of seminal leukocytes in routine semen analysis appears to be of little prognostic value with regard to male fertilizing potential. As reactive oxygen species and antisperm measurement were of similar predictive value, the term "immunologic male infertility" should be redefined. PMID- 8243689 TI - Assessment of human sperm acrosome reaction by flow cytometry: validation and evaluation of the method by fluorescence-activated cell sorting. AB - OBJECTIVE: To determine the applicability of flow cytometry to assess human sperm acrosome reaction. DESIGN: Prospective evaluation of semen samples incubated overnight for the development of spontaneous acrosome reaction or exposed to the calcium ionophore A23187 (5 microM) for 3 hours for induction of the acrosome reaction. SETTING: University-affiliated tertiary care center. PATIENTS: Normal healthy volunteers. INTERVENTIONS: The spermatozoa were stained with fluorescein isothiocyanate (FITC)-labeled pea agglutinin. The labeled samples were assessed visually and also subjected to analytic flow cytometry and fluorescence-activated cell sorting. MAIN OUTCOME MEASURES: Acrosome reaction assessed visually and by flow cytometry. RESULTS: Flow cytometric analysis showed a single peak of FITC fluorescence in the washed semen samples. A second peak of lower FITC fluorescence intensity was noted after overnight incubation or exposure to A23187, suggesting loss of fluorescence, which indicated the occurrence of the acrosome reaction. There was a statistically significant correlation between the assessment by the two methods (n = 41). However, although the mean difference between the methods was small (2.49%), the difference between the two methods for each individual sample can vary between -24% to +29%. When the sperm cells were subjected to cell sorting based on green fluorescence intensity, reanalysis and visual scoring verified that the low intensity peak contained a majority of acrosome-reacted spermatozoa (77.52% +/- 2.39%). CONCLUSION: These results validated the flow cytometric method for assessment of acrosome-reacted spermatozoa. Although flow cytometry is more objective and less time consuming when many samples are assessed at the same time, the visual method remains a useful and practical procedure in the clinical andrology laboratory. PMID- 8243690 TI - Numbers and quality of oocytes after induction of multiple folliculogenesis in women and in mice with different lots of human menopausal gonadotropins. AB - OBJECTIVE: To develop a murine in vivo bioassay that is sensitive to inherent variability of hMG. DESIGN: Female F1 hybrid mice were mated after injection with fixed dosages of each of four lots of hMG. Oocytes/embryos were collected and monitored during 5 days of culture. Outcome measures were compared with retrospective outcomes of ovarian stimulation of women with the same hMG lots. The bioassay was then used to compare biopotencies of six different lots of hMG. SETTING: Center for assisted reproductive technology. INTERVENTIONS: None. PATIENTS: Outcomes of controlled ovarian stimulation and IVF were retrospectively analyzed from 43 patients. MAIN OUTCOME MEASURES: Numbers of oocytes/embryos recovered, numbers fertilized, and numbers progressing beyond fertilization. RESULTS: Differences in biopotencies of hMG lots in the murine in vivo bioassay were consistent with differences in outcomes of patients treated with the same hMG lots during controlled ovarian stimulation. Bioassay outcomes also differed between different lots of hMG. CONCLUSION: The bioassay described in this study is sensitive to between-lot differences in hMG that affect follicle recruitment and oocyte quality in women (and in mice). In this respect, the bioassay could be of value in screening hMG lots, before its exploitation in a patient population. One such application is explored. PMID- 8243691 TI - The developmental potential of mouse embryos conceived in Ham's F-10 medium containing ethylenediaminetetraacetic acid. AB - OBJECTIVE: To measure fetal developmental potential of mouse eggs fertilized and cleaved in modified Ham's F-10 medium (Sigma Chemical Co., St. Louis, MO) containing the chelating agent, ethylenediaminetetraacetic acid (EDTA), as a model for establishing criteria for human IVF and GIFT procedures. DESIGN: Optimum concentrations of EDTA in modified Ham's F-10 were established by measuring blastocyst development of in vivo fertilized zygotes from a random bred (CD1) mouse strain. Postimplantation development of in vitro fertilized eggs from a hybrid inbred (B6D2F1) mouse strain was measured after conception in the modified Ham's F-10 medium without or with human serum or EDTA and compared with the results of in vivo fertilized eggs. Mouse embryo development potential and pregnancy rates (PRs) of recipients were compared with those calculated from theoretical probabilities. RESULTS: The addition of 10 to 100 microM EDTA to modified Ham's F-10 supported development of > 60% of the in vivo conceived zygotes to the blastocyst stage in the absence of serum; the addition of human serum significantly decreased embryo development to blastocysts (23%). Pregnancy rates (30% to 50%) of recipients of the in vitro conceived mouse embryos were similar for embryo cohorts from all culture conditions tested. The overall yield of fetuses from total embryos transferred was significantly higher for the groups developed in modified Ham's F-10 with 100 microM EDTA (37%) compared with embryos developed in modified Ham's F-10 alone (18%) or in modified Ham's F-10 with human serum (18%). CONCLUSIONS: [1] Based on fetal development in pregnant recipients, approximately two thirds of the mouse eggs fertilized in modified Ham's F-10 with 100 microM EDTA (or in vivo) had the potential to give rise to offspring. [2] Given the viability rate, the observed PRs of foster mothers receiving four to six embryos were only two thirds of the theoretically possible, suggesting the hormone and/or surgical interventions of the recipients reduced their pregnancy potential. [3] Including 100 microM EDTA during mouse conception in modified Ham's F-10 improved the fetal developmental potential to that of in vivo derived mouse embryos. PMID- 8243692 TI - Electroejaculation using standard nerve stimulation equipment and Teflon-coated needles. AB - We attempted transperineal needle electroejaculation using a Digistim nerve stimulator and Teflon-coated needles in 12 anejaculatory men. We obtained semen in 11 men. Five of the 12 men also underwent electroejaculation using a transrectal probe. Comparable semen parameters were obtained by transrectal probe and transperineal needles. PMID- 8243693 TI - Dealing with uncertainty--the challenge of the decade? PMID- 8243694 TI - Coculturing--the way to blastocyst transfers? PMID- 8243695 TI - Physiopathology of spermatogenic arrest. AB - OBJECTIVE: To review the world literature on the etiology, physiopathology, and treatment of spermatogenesis arrest. STUDY SELECTION: All the pertinent literature on spermatogenic arrest has been selected. Most studies related to this topic have been identified through Medline and through published literature. PATIENTS: Spermatogenic arrest has been diagnosed by testicular biopsy in men of reproductive age who had either severe oligospermia (partial arrest) or azoospermia (complete arrest), normal testicular volume, and depending on the etiology normal, high, or low levels of gonadotropins. INTERVENTIONS: The effects of heat, radiotherapy, and chemotherapy have been reported. Depending on the etiology of spermatogenic arrest, different hormonal treatments have been tested. MAIN OUTCOME MEASURE: Level of interruption of germ cell differentiation in testicular biopsy have been determined. Improvement of the sperm count or appearance of mature sperm after an hormonal treatment have been observed. RESULTS: Spermatogenic arrest can occur at spermatogonial level in case of gonadotropin insufficiency or after germ cell damage due to chemotherapy or radiotherapy. The arrest is most frequently observed at primary spermatocyte level. Reversible arrest at that level can be due to heat, infections, hormonal and nutritional factors. Irreversible arrest at primary spermatocyte or spermatid level have a genetic origin due to chromosomes anomalies either in somatic cells or in germ cells. CONCLUSIONS: Spermatogenic arrest is usually due to genetic factors resulting in irreversible azoospermia. However some cases may be consecutive to hormonal, thermic, or toxic factors and may be reversible either spontaneously or after a specific treatment. PMID- 8243696 TI - Spermatogenic arrest: a classical and futuristic view. PMID- 8243697 TI - Endometriosis and pelvic pain. PMID- 8243698 TI - Management of endometriosis in the presence of pelvic pain. The American Fertility Society. PMID- 8243699 TI - The effect of operative technique and uterine size on blood loss during myomectomy: a prospective randomized study. AB - OBJECTIVES: To compare operative blood loss between two accepted blood loss reducing techniques used during myomectomy and to evaluate the effect of preoperatively determined uterine volume on blood loss. DESIGN: Subjects were stratified by ultrasound-determined uterine volume < 600 cm3 (n = 11) and > or = 600 cm3 (n = 10) and then randomized into treatment groups. The same radiologist, surgeons, and anesthetic induction technique were involved in every case. In the pharmacologic technique, diluted vasopressin (20 U in 20 mL normal saline) was injected into the serosa and/or myometrium overlying the fibroid(s) before the uterine incision(s). In the mechanical technique, a penrose drain tourniquet was passed through defects created in the broad ligaments at the level of the internal os and secured posteriorly, occluding the uterine vessels. In addition, vascular clamps were placed on the infundibulopelvic ligaments, occluding anastomotic blood flow through the ovarian vessels. RESULTS: There was no difference in operative blood loss, operating time, preoperative and intraoperative mean arterial blood pressures, postoperative febrile morbidity, preoperative and postoperative hematocrits, transfusion rates, and length of hospital stay between groups. Blood loss was significantly greater for uteri with ultrasound-determined volumes > or 600 cm3 (627 +/- 175 mL, mean +/- SEM) than for those < 600 cm3 (228 +/- 49 mL). For all subjects, blood lost while operating on the uterus (mean, 379 mL; range, 35 to 1,968 mL) was positively correlated with the total weight of the fibroids resected and with time spent operating on the uterus. Total blood loss (mean, 418 mL; range, 42 to 1,968 mL) was also positively correlated with the time spent operating on the uterus and with total operating time. CONCLUSIONS: There were no demonstrable differences in blood loss, morbidity, or transfusion requirements between subjects undergoing myomectomy using pharmacologic vasoconstriction and mechanical vascular occlusion techniques. Blood loss during myomectomy is primarily incurred while operating on the uterus and is correlated with preoperative uterine size, total weight of fibroids removed, and operating time. PMID- 8243700 TI - Cost of ectopic pregnancy management: surgery versus methotrexate. AB - OBJECTIVE: To estimate the potential annual cost savings of methotrexate (MTX) therapy for ectopic pregnancy (EP) at our institution and nationally. DESIGN: Retrospective. SETTING: San Francisco General Hospital. PATIENTS: All patients treated for EP in 1991. INTERVENTIONS: Medical records and billing statements were reviewed. MAIN OUTCOME MEASURES: Direct costs were determined from billing statements, and indirect costs were estimated based on published data. Potential annual national cost savings from MTX treatment were then extrapolated using national statistics on the incidence of EP. RESULTS: Fifty EPs were treated with an average direct cost of $7,972 per case. Direct and indirect costs totaled > $500,000. Fifteen (30%) of the cases would have been eligible for MTX treatment with a resultant cost savings of > $160,000. The potential annual national cost savings would be in excess of $280 million. CONCLUSIONS: If preliminary findings suggesting the effectiveness of MTX as an alternative therapy for EP are confirmed, appropriate use of MTX will not only reduce morbidity but also the substantial cost. PMID- 8243701 TI - The association between Chlamydia trachomatis serology and pelvic damage in women with tubal ectopic gestations. AB - OBJECTIVE: To determine whether pelvic damage is associated with positive Chlamydia trachomatis serology in women with tubal ectopic pregnancy. DESIGN: Cross-sectional retrospective study. SETTING: A prepaid health maintenance organization. PATIENTS: Two-hundred eighty-one women admitted with confirmed tubal ectopic pregnancy were interviewed for history of sexually transmitted diseases. Chlamydia serology was obtained for 135 subjects, and operative findings were available for 121 of these. INTERVENTIONS: None. MAIN OUTCOME MEASURE: Pelvic damage, as determined by review of operative findings of the pelvis at the time of ectopic surgery. RESULTS: Pelvic damage was associated with positive chlamydia serology with an adjusted odds ratio of 4.2 (95% confidence interval: 1.8 to 9.7). Moderate and severe pelvic damage were more strongly associated with positive serology than mild damage. CONCLUSIONS: Women with ectopic pregnancies and antibodies to C. trachomatis are more likely to have damaged pelves than women with ectopic pregnancies without such antibodies. Prevention or early treatment of C. trachomatis infection may reduce pelvic damage and, therefore, reduce incidence of ectopic pregnancy. PMID- 8243702 TI - Long-term follow-up of the twisted ischemic adnexa managed by detorsion. AB - OBJECTIVE: To examine our assumption that although the twisted adnexa appears ischemic-hemorrhagic, it can safely be revived by detorsion with preservation of ovarian function. DESIGN: Patients' records were obtained from a computerized database and reviewed. The preoperative diagnosis of adnexal torsion was based upon patients' symptoms, clinical examination, laboratory investigations, and ultrasound scanning. SETTING: Gynecology department at a large teaching hospital receiving primary referrals of public sector patients. PATIENTS: In 40 patients who presented with signs and symptoms suggestive of ovarian torsion, "black bluish" ischemic adnexa were encountered at surgery. INTERVENTIONS: All patients were managed by unwinding of the adnexa: laparotomy in 26 cases and operative laparoscopy in 14. In 13 patients detorsion only was performed, in 15 detorsion and cystectomy were carried out, and in 12 patients detorsion was done and ovarian cysts aspirated. MAIN OUTCOME MEASURES: Postoperative course, mean hospitalization period, follow-up pelvic examination, ovarian folliculogenesis on ultrasound examination, and oocyte retrieval and fertilization. RESULTS: The size of the twisted ovary ranged from 4 to 20 cm (mean, 9.5 cm). The postoperative course was uneventful, except for transient temperature elevation in five patients. The mean hospitalization period was 6.5 days (range 5 to 10 days) after laparotomy and 1.8 days (range 1 to 3 days) after laparoscopy. Three patients were lost to follow-up. In 37 patients, pelvic examination was normal. A normal sized ovary, with follicular development, was demonstrated sonographically in 35 of 37 patients. In 6 of 7 patients, macroscopically normal adnexa were visualized at subsequent laparotomy or laparoscopy. In two patients undergoing IVF, oocytes were retrieved and fertilized from the detorted ovary. The patency of the fallopian tube was demonstrated in four cases. CONCLUSIONS: This new "adnexal sparing" approach should be applied instead of the traditional salpingo oophorectomy in young women with twisted ischemic adnexa. PMID- 8243703 TI - Gonadotropin-releasing hormone agonist administration in early human pregnancy is associated with normal outcomes. AB - OBJECTIVE: To evaluate the reproductive outcomes of early human gestations exposed to GnRH agonist (GnRH-a). Gonadotropin-releasing hormone agonist is commonly used as an adjunct to ovulation induction regimens, and sometimes early pregnancies are inadvertently exposed to the drug. There has been concern regarding possible teratogenic effects of GnRH-a on these gestations, but there is a paucity of data regarding human subjects. DESIGN: Identified pregnancies were followed for obstetric and fetal outcomes. PATIENTS, PARTICIPANTS: Over 4 years 18 early pregnancies among 17 women were found to be exposed to GnRH-a. During the month of conception, daily injections were begun on menstrual day 22 after documenting an ovulatory P level on day 21. Gonadotropin-releasing hormone agonist was discontinued from 10 to 24 days later, immediately after pregnancy was recognized. Obstetric and fetal outcomes were ascertained by chart review and telephone follow-up with mothers. RESULTS: Five first trimester losses were observed in 18 hCG confirmed pregnancies for a loss rate of 28%. Thirteen pregnancies produced apparently normal babies. One normal fetus was lost at 20 weeks' gestation because of cervical incompetence. CONCLUSION: There does not appear to be an increased risk of birth defects or pregnancy wastage in human pregnancies exposed to daily low-dose GnRH-a therapy in the first weeks of gestation. PMID- 8243704 TI - Diminished growth hormone responses to L-dopa in polycystic ovarian disease. AB - OBJECTIVE: To investigate the GH secretion capacity in polycystic ovarian syndrome (PCOS). DESIGN: Prospective. SETTING: Reproductive endocrinology unit. PATIENTS: Nineteen women with clinical and endocrinologic signs of PCOS were compared with 10 healthy weight-matched volunteers with normal menstrual rhythm and no history of endocrine or reproductive abnormality. INTERVENTIONS: Standard GH reserve estimates were affected in all subjects using oral L-Dopa. The test was carried out after an overnight fast on cycle days 5 to 8 in menstruating subjects; the test was done randomly in patients with amenorrhea. RESULTS: Both PCOS and control groups responded to L-Dopa over a 3-hour period with significant elevations in GH concentrations. The control group showed significantly higher values than the PCOS group at 90, 120, and 180 minutes. CONCLUSION: The findings indicate that PCOS patients demonstrate a relative deficiency in GH reserve. PMID- 8243705 TI - Counter-regulatory response to hypoglycemia in the follicular and luteal phases of the menstrual cycle. AB - OBJECTIVE: To assess whether the phase of the menstrual cycle influences the counter-regulatory response to hypoglycemia. DESIGN: Prospective randomized euglycemia-hypoglycemia clamp studies in the follicular and luteal phases of the menstrual cycle in the same woman. SETTINGS: Clinical Research Center, Yale University School of Medicine. PATIENTS: Eight regularly menstruating nonobese women. MAIN OUTCOME MEASURES: Counter-regulatory hormonal response to hypoglycemia-epinephrine, norepinephrine, glucagon, cortisol, GH, and PRL; glucose homeostasis: rates of whole-body glucose appearance and utilization, and the rate of hepatic glucose production. RESULTS: In the follicular and luteal phases of the cycle fasting glucose levels (88 +/- 1 and 85 +/- 2 mg/dL, mean +/- SEM, respectively; conversion factor to SI units, 0.05551), basal glucose turnover (2.37 +/- 0.20 and 2.63 +/- 0.13 mg/kg per minute), basal insulin levels (10 +/- 1 and 9 +/- 1 microU/mL; conversion factor to SI units, 6.0), and insulin levels during the clamp study (53 +/- 3 and 45 +/- 4 microU/mL) were not significantly different. During the euglycemic phase of both studies, glucose utilization rose twofold (to 4.73 +/- 0.31 and 4.39 +/- 0.31 mg/kg per minute): hepatic glucose production was suppressed; and counter-regulatory hormones remained unchanged. Induction of hypoglycemia produced increases in the concentrations of counter-regulatory hormones that were indistinguishable in both phases of the cycle. Similarly, the increase in hepatic glucose production provoked during hypoglycemia was similar in each phase of the cycle (1.20 +/- 0.24 and 1.28 +/- 0.36 mg/kg per minute). CONCLUSION: The counter-regulatory hormonal response to hypoglycemia, as well as the metabolic sequelae of these hormonal changes, are similar in the follicular and luteal phases of the menstrual cycle. PMID- 8243706 TI - Differential effects of oral glucose-mediated versus intravenous hyperinsulinemia on circulating androgen levels in women. AB - OBJECTIVE: To determine whether transient endogenous or continuous exogenous hyperinsulinemia produced different changes in circulating gonadal steroid levels because insulin is a putative regulator of gonadal steroid secretion in polycystic ovarian syndrome (PCOS). DESIGN: An oral glucose tolerance test (OGTT) and a 2-hour euglycemic clamp (+100 microU/mL [718 pmol/L] steady-state insulin levels) were performed in obese patients with PCOS (n = 7) in nonobese patients with PCOS (n = 5), and in age- and weight-matched ovulating normal women (obese normal n = 7, nonobese normal n = 6). RESULTS: Despite increased insulin levels, the levels of T and androstenedione (A) decreased in three of the four groups during the OGTT (nonobese normal women showed a slight increase in T during the OGTT). In contrast, the continuous 2-hour insulin infusion resulted in increased androgen levels. When all four groups were pooled, the difference in the changes in A levels between the two tests was significant, and the difference in T levels between the two tests approached significance. CONCLUSIONS: Transient physiological hyperinsulinemia produced by an oral glucose load was associated with a decrease rather than an increase in circulating androgen levels. The effects on circulating androgen levels of oral glucose-mediated increases in insulin levels were significantly different from those of a continuous intravenous insulin infusion. Sustained hyperinsulinemia produced by exogenous insulin infusion appeared to be required to increase androgen levels in women; transient physiological increases in insulin levels after an oral glucose load were insufficient to produce hyperandrogenism. Postmeal hyperinsulinemia does not contribute to hyperandrogenism in women. PMID- 8243707 TI - [Effect of liposomes on tissue respiration in animals with acute hypoxic hypoxia]. AB - Introduction of liposomas has been studied in experiments on Wistar rats for its effect on oxygen demand by tissues of the brain great hemispheres, lungs, liver and femoral muscle under acute hypoxic hypoxia due to inhalation by an animal of gas mixtures containing of 6.7-7.1% of O2. It is shown that after introduction of phospholipid liposomes as a lipin preparation against a background of acute and grave forms of hypoxic hypoxia the oxygen demand of tissues remains practically at the level of control values. This proves absence of the tissue oxygen debt that usually develops at hypoxic states. It is concluded that the organism tissues in the presence of liposomas do not practically suffer from the oxygen deficiency and introduction of lipin may be effective to prevent from development of the secondary tissue hypoxia. PMID- 8243708 TI - [The role of bile acids in the regulation of human gallbladder filling]. AB - An artificial increase of a pool of bile acids due to administration of dehydrocholic, chenodesoxycholic acid preparations and dry bile in 22 healthy people and 179 patients with chronic cholecystitis has been studied for its effect on regulation of the gallbladder filling on an empty stomach using daily echocholecystometry. Filling of the gallbladder under artificial sequestration of bile acids during duodenal probing, enterosorption and cholatogenic diarrhea was studied in 55 patients with cholecystitis. It has been proved that an increase of the pool of bile acids induces intensification of the gallbladder filling on an empty stomach, while a decrease of the pool, vice versa, causes attenuation of its filling. It is found out a size of a pool of bile acids in the human organism is an important humoral factor in the physiological mechanism of the gallbladder filling regulation. PMID- 8243709 TI - [Mechanisms of the negative effect of blood serum on iodine-131 uptake by cultured thyrocytes]. AB - Embryonal (calf) and ordinary (cattle) blood sera have ben studied experimentally on a primary culture of thyroid cells from newborn pigs for their effect on basal and thyrotropin (TSH)-stimulated 131I uptake by thyrocytes. It is shown that the value of this index is inversely proportional to the relative percentage of the blood serum in the cultivation medium. The concentrations being equal, the ordinary blood serum effect is more pronounced than the embryonic one. Thyrocyte incubation in a medium with 1% blood serum and 0.1-10.0 mU/ml TSH stimulates 131I uptake by the cells, whereas 15% concentration of the blood serum blocks the influence of this hormone. Ouabain (0.1 mM) and cyclohexamide (10 micrograms/ml) have also an inhibiting effect on exposure of the basal and TSH-stimulated uptake of the isotope. It is suggested that the blood serum contains biologically active substances inhibiting the transmembranous transfer of iodides into thyrocytes. This process is controlled by Na+, K(+)-ATPase, and its TSH-stimulation is mediated by protein synthesis de novo. PMID- 8243710 TI - [Changes in concentration of the blood serum malondialdehyde due to a one-time and long-term ethanol and acetaldehyde administration]. AB - Changes in concentration of the rat blood serum malone dialdehyde (MDA) at separate and combined action of ethanol and acetaldehyde under conditions of their single and long-term administration have been investigated. In some series of experiments acetaldehyde and ethanol were administered together with teturame. The obtained data permit supposing that under conditions of acute ethanol intoxication an increase of the MDA concentration is caused by the acetaldehyde action. The long-term ethanol intoxication induces a significantly higher (if compared with acute intoxication) increase the MDA concentration. The results confirm the concept of the lipid peroxidation process at acute and chronic intoxication due to ethanol metabolism. Acetaldehyde is an important factor among those determining disturbances of cell biogenesis in the animal organism, causing an increase of the blood serum MDA. PMID- 8243711 TI - [Activation mechanisms of lipid peroxidation in rat's tissue in various types of hypoxia]. AB - The mechanisms of lipid peroxidation (LP) intensification in the rat myocardium, brain and liver were studied under hypoxic hypoxia and hemic hypoxia. The level of intensification depends on the type of hypoxia and is characterized by tissue specificity. It is shown that LP is activated by hypoxia. This activation is more pronounced under hemic hypoxia associated with more marked catecholamine accumulation, deficiency of the antioxidant glutathione system, decrease of the cytochrome c-oxidase activity. The level of lipid peroxides was higher in the rat liver under both types of hypoxia. PMID- 8243712 TI - [Effect of 6-hydroxydopamine administration to newborn female rats and sex differentiation of the brain]. AB - Remote effects of neonatal 6-hydroxydopamine (6-OHDA) combined with testosterone propionate (TP) and alone were compared to ultrastructure of catecholaminergic terminals of the median eminence and hypothalamic catecholamine (CA) content in the critical period of sexual differentiation of the brain. It is suggested that inability of 6-OHDA to prevent TP-induced anovulatory sterility is connected with the preserved hypothalamic CA levels and a lack of significant degenerative changes in CA-ergic terminals in the median eminence. PMID- 8243713 TI - [Distribution of 1-[14C] GABA and its conjugates with nicotinate, pyridoxal phosphate and biotin in tissues of guinea pigs in closed systems]. AB - Distribution of the total radioactive label in the blood, different parts of the brain (the whole cortex and its lobes, bulbi olfactorii, hippocamp, striopallidum, hypothalamus, thalamus, peduculi cerebri, cerebellum, pons, medulla oblongata), eye (retina, hyaloid and vascular membrane), hypophysis and liver of guinea pig males has been studied after the hypodermic injection of [14C] GABA and products of its conjugation with pyridoxal phosphate, nicotinic acid and biotin in the equimolar dose (50 nmol/g) under normal conditions and in case of action of the closed volume factors. Accumulation of the total label in the hypophysis after injection of [14C] GABA, PLP-[14C] GABA and nicotinoyl-[14C] GABA was 3.6, 3.1 and 2.7 times (respectively) as much as that in the blood. The content of the labelled compounds in the cortical structures of the telencephalon and in the diencephalon was the greatest in comparison with other parts of the brain. The permeability through a hemato-ophthalmic barrier was 3-5 times as much as that through a hemato-encephalic barrier. The closed volume factors lead to higher accumulation of the label as compared with the control after injection of [14C] GABA, PLP-[14C] GABA and biotinyl-[14C] GABA. PMID- 8243714 TI - [Respiratory reactivity and parameters of nonspecific immunity in workers of the Chernobyl nuclear power plant during adaptation to mountain climate]. AB - The quantitative composition of the peripheral blood, enzymatic activity of neutrophils and hypoxic ventilatory drive were determined in people working at the Chernobyl NPP during sojourn for 28 days in the Caucasus mountains at 220 m altitude with a periodical climbing up to 4200 m. At the initial state we found neutropenia, lymphocytosis, an increase in the number of prolymphocytes and big hairy lymphocytes, a decrease in the number of small lymphocytes, fall in activity of enzymes responsible for oxygen-dependent and oxygen-independent mechanisms of bactericidity. The indices of sensitivity to the hypoxic respiration stimulus did not essentially differ from the norm. Staying at altitude promoted normalization of the number of segmento-nuclear neutrophils, lymphocytes (their small population, in particular), an increase in the number of eosinophiles of the peripheral blood. Changes in the activity of myeloperoxidase, NADP-oxidase and cationic proteins in the neutrophils were observed. A considerable increase of the ventilatory drive to the hypoxic respiration stimulus was determined. Accentuation of fermentative transformations proved to be closely related to the ventilatory response to hypoxia: weak activation of NADP-oxidase and a decrease of myeloperoxidase activity were observed in people with initially low respiration reactivity, while a considerable increase in activity of these enzymes and in content of cationic proteins was observed in people with high respiration reactivity. Individual peculiarities of the reaction to staying at altitude should be taken into account when developing particular methods of immune correction. PMID- 8243715 TI - [Oxygen-dependent processes in the liver and immunologic reactivity of rats in chronic liver damage induced by carbon tetrachloride]. AB - The chronic hepatitis in Wistar rats was induced by seven subcutaneous injections of carbon tetrachloride (0.3 ml of 50% oil solution per 1 kg of body mass) made each third day. It was shown that cytochromes P-450 and b5 content, demethylase activity in the liver microsomal fraction as well as the oxygen tension in the liver tissue were decreased, while lipid peroxidation was intensified. The PHA induced blood lymphocyte blastogenesis was inhibited and concentration of circulating immune complexes in the blood was higher than that in intact animals. PMID- 8243716 TI - [Role of catecholamines, released due to stress, on stimulation of heparin secretion by mast cells in rats]. AB - The secretory status of mast cells of mesentery and subcutaneous connective tissue was studied in rats using the morphometry analysis. Immobilization for 30 min has induced a 3.7-fold decrease of the heparin saturation index in these mast cells and a 3-fold increase of the granulolysis index. In rats preliminary given propranolol, alpha-adrenoceptor antagonist (1.25 mg/kg), a stimulatory effect of immobilization was absent. Phentolamine, alpha-adrenoceptor antagonist (6 mg/kg), partially inhibited the immobilization effect. Isoprenaline, beta-agonist (1.5 mg/kg), activated heparin secretion in rats without immobilization, while phenylephrine, alpha-agonist (2.5 mg/kg) had no effect on the secretory state of mast cells. Propranolol in the same dose also blocked the stimulatory action of i. v. injected alpha-thrombin (50 NIH/kg) on heparin secretion from mast cells of subcutaneous connective tissue. The obtained results show that catecholamines participate in activation of heparin release from mast cells induced both by the stress and by thrombin injection. Their action is mediated by beta-adrenoceptors of the vessel wall. PMID- 8243717 TI - [Effect of human colostrum and its fraction on functional activity of B lymphocytes]. AB - The human colostrum stimulated by suboptimal doses (50 ng/ml) of lipopolysaccharide (LPS) has induced proliferation of mouse splenocytes. Addition of the human colostrum has elicited a rise in splenocyte culture supernatants. Effect of human colostrum on Ig synthesis was dose-dependent and was expressed at 0.5% concentrations in the medium. In contrast to the human colostrum, the bovine colostrum exerted a suppressive effect on proliferation of mouse splenocytes and Ig synthesis induced by suboptimal doses of LPS. Bovine casein did not significantly affect the Ig synthesis by mouse splenocytes. Suppression of the Ig synthesis was associated with bovine colostral globulins. Using the ELISPOT method we have found that a rise in the Ig level in the culture after treatment with the human colostrum was due to an increase of the number of IgG2a-secreting cells. B-cell promoting fraction was found after HPLC gel-filtration. The molecular weight of this fraction was 70-75 kD. PMID- 8243718 TI - [Protective effect of Lactobacillus acidophilus on development of infection, caused by Klebsiella pneumoniae]. AB - The mechanisms of protective action of Lactobacillus have been studied during development of the generalized infection induced by Klebsiella pneumoniae in CBA mice after weaning. The mice were infected intragastrically during the first day after weaning (1 x 10(9) bacterias per mice). Suspensions of Lactobacillus were introduced before and after infection during 10 days (1 x 10(6) bacterias per mice). It has been shown that introduction of Lactobacillus substantially decreased the level of the gut contamination by Klebsiella, prevented generalization of infection and death of animals. Significant higher levels of IgA in the blood serum, IgA and IgM in the gut content, percentage of splenocytes, expressing surface IgM and IgG were observed on the 7th day as compared with those in animals without Lactobacillus. Significantly lower percentage of splenocytes, expressing CD4 antigen was also observed. On the 11th day after infection the mice receiving lactobacillus have shown a tendency to an increase of IgA in the gut content, significantly lower concentrations of IgM in the gut content and a higher level of IgA to the blood serum as compared with the control. Other characteristics were comparable to those of the control group. A conclusion is made that introduction of Lactobacillus prevents development of the Klebsiella infection and protects the immune system from excessive antigenic action. PMID- 8243719 TI - [Formation of immunologic memory to antigens of tuberculosis mycobacteria in mice]. AB - The antibody immune response and development of immunological memory to Murobacterium bovis antigens were studied in experiments carried out on CBA and BALB/c mice. Optimal antigen does for primary immune responses were 1 mg of both live and killed bacterial cells. In contrast to the primary antibody production, formation of the immunological memory was found to depend greatly on the viability of M. bovis injected. The levels of circulating immune complexes increased during secondary antibody responses demonstrated only in mice primed with live mycobacteria but not with killed ones. Aggregation of a purified derivative of tuberculin protein by crosslinking polypeptides using chromium chloride resulted in a significant enhancement of their immunogenic properties providing the high level of immunological memory formation as well as production of the antibodies against M. bovis specific antigens. PMID- 8243720 TI - [Effect of triiodothyronine and isopropyl noradrenaline on the "set-point" temperature in white rats]. AB - It has been shown that in experiments where a negative feed-back method has been used, triiodothyronine and beta-adrenomimetic isopropylnoradrenaline exert a significant influence on the work of the whole system of thermoregulation in white rats. This is observed in an upward displacement of the "set-point" temperature. PMID- 8243721 TI - [Correlation of basic properties of nervous processes with volitional attention in young school children]. AB - It is revealed that children 7-9 years old lack a correlation between properties of principal nervous processes and properties of attention. Such connection appears only in age of 9-10. Development of properties of principal nervous processes as well as of attention in early ontogenesis is characterized by their quick growth. PMID- 8243722 TI - [Effect of experimental hypokinesia on adrenergic reactivity of the cardiovascular system in rabbits]. AB - In the experiments on rabbits under prolonged hypokinesia the functions of the cardiovascular system and the adrenergic reactivity of blood pressure were studied. The increase of the heart rate at rest, clear tendency to the decrease of the arterial blood pressure and no changes of the venous pressure are shown. The reliable decrease of the maximum size and duration of the blood pressure reactions to adrenaline are demonstrated. These changes prove the decrease of the functional possibilities and adrenergic reactivity of the cardiovascular system under hypokinetic conditions. PMID- 8243723 TI - [Comparative study of nerve stimulation factors by a single rectangular impulse and a series of rectangular impulses]. AB - The experiments were performed on the sciatic nerve of Rana temporaria. A single rectangular electric impulse and a series of rectangular microsecond impulses were used for stimulation. The threshold amplitude of a single impulse and the average voltage in series of impulses were compared. It was experimentally shown that application of a series of impulses as a stimulus necessitated less values of stimulation near the threshold. There was no significant difference between the average voltage in a series of impulses and the amplitude of a single rectangular impulse in stimulation far up the threshold. PMID- 8243724 TI - [Changes in energy metabolism parameters of the liver in acute fluoride intoxication and hyperbaric oxygenation]. AB - Examinations of energetic metabolism in liver of white rats with acute fluoride intoxication were carried out in acute and recovering periods. The influence of hyperbaric oxygenation on the change of energetic metabolism indices and on survival rates of animals has been studied. Hyperbaric oxygenation greatly prevented profound metabolic disturbances, in particular, ATP, increased detoxic in liver intensifying energetic forming processes. The survival rate of animals under the influence of course of HBO increased by 27% during first 72 hours. It was determined that HBO has an effective influence on the course of recovering period during fluoride intoxication. PMID- 8243725 TI - [Changes of various physiologic characteristics in adult rats as effected by electrostatic field]. AB - The chronic effect of electrostatic field has been studied in experiments on adult female Wistar rats. It is shown, that under these conditions the stability of erythrocyte membranes decreases, that causes their earlier hemolysis. The level of calcium and iron in blood increases, that is correlated with changes in the urea content. In the brain iron content decreases, while urea increases. The obtained results reflect the active establishment of dynamic equilibrium in conformity with this influence. PMID- 8243726 TI - [Effect of caffeine on chronotropic correlations in the myocardium of the rat and guinea pig]. AB - Force-frequency relations in the isolated papillary muscles of rats and guinea pigs were compared using isometric force measurement technique. Stimulation frequency varied between 0.33 and 4 Hz. Under normal conditions the rat papillary muscle exhibits a negative force-frequency dependence which differs from ventricular preparations in many other mammals. Caffeine (10 mmol/l) introduced into the bath solution abolishes a negative force-frequency dependence in the papillary muscle of rat. During incubation of the rat ventricular preparations in the caffeine-containing solution the force-frequency relations measured on these muscles display the same behaviour as in guinea pig preparations. Caffeine has induced no changes in the force-frequency relations of guinea-pig ventricular preparations. A conclusion is made that caffeine-sensitive intracellular calcium stores participate in species-determined differences in force-frequency relations of the ventricular muscles. PMID- 8243727 TI - [Closure of cleft palate in adult patients, using a forearm flap with microvascular radial artery II]. AB - A clinical report on the utilization of the so called "Chinese Flap" first published in 1981 is presented by the authors. The basic principle of this method is the following: After closing the soft palate in three layers, a mucoperiosteal flap has been prepared from the palatal mucosa and rotated into the nasal cavity so to close nasally the defect in one layer. Than a pediculated fasciocutaneous flap containing both the radial artery and the two comitantes veins prepared from the forearm was sutured to the defect. The a radialis and the v. comitantes running forward and downward between the two layers of the soft palate passing the mandibular ramus and descending through a tunnel leading to the submandibular region were joined to the facial artery and veins and an anastomosis was established. The clinical experience with the "Chinese Flap" procedure showed good results. This method has been used in 12 cases so far and only one out of the 12 failed due to blood vessel developmental abnormalities. PMID- 8243728 TI - [Mechanism of the eruption of secondary dentition]. AB - The second dentition is a process determined in space and time which presumes the presence of the permanent dental germ, its optimal position and the soundness of the periapical area. In absence of these conditions the clinically and histologically proved pathological phenomena (ex. the pathological triad of second dentition) can occur. PMID- 8243729 TI - [Heredity as an etiologic factor in dentistry (oral genetics)]. AB - The oral genetics is the part of the stomatology that is related to the questions of heredity. The oral genetics searches the typical morphological characteristics, the congenital anomalies, other pathological alterations and diseases, as well as hereditary syndromes. The author describes the genetical relevance of the two most important oral diseases: caries and parodontopathy. PMID- 8243730 TI - [Hormonal regulation of bone metabolism]. AB - During the past two decades our ideas about the regulation of bone formation and resorption has been expanded. At the cellular level we have learnt more about the hormonal control of the osteoblastic bone formation, the osteoclastic bone resorption and the role of the nonosseous cells in the bone metabolism. At the tissue level we have understood the coupling between osteoblastic bone formation and osteoclastic bone resorption and consequently the relation between bone turnover and skeletal growth, involution and pathologic bone changes. Though many of the findings of the recent bone researches have helped us to understand the pathogenesis of the metabolic and infectious bone diseases we do not yet have a reliable means for stimulating bone formation of structurally and functionally effective new bone in human beings. In this review we tried to summarize the research data and clinical observations showing the direct and indirect effect of calcium regulating hormones (PTH, 1,25 dihydroxyvitamin D and calcitonin) on bone metabolism. PMID- 8243731 TI - [Corrosion of NiCr- and CoCr-containing dental alloys in cellulose-based artificial saliva under in vitro conditions]. AB - It was found that the CoCr alloy is more resistant to the corrosive effects than NiCr alloy. The artificial saliva developed at our clinic increased the passivation range of the both alloys. The higher value observed in the case of the CoCr alloy makes it more suitable for use in patients with xerostomia or an increased risk of allergy. PMID- 8243732 TI - [Electrochemical properties of dental alloys]. PMID- 8243733 TI - [History of the Hungarian Dental Association]. AB - The Hungarian Dental Association was founded in 1878. Its history can be divided into three periods. The first was the Austro-Hungarian Monarchy until the end of the 1st World War. The second was after the Trianon peace- pact during the Horthy regime, the third during the Soviet occupation. The fourth period starts now, in the--for the first time--independent, free Hungary. PMID- 8243734 TI - [Use of hydroxyapatite (Ceros-80) in dental implantology]. AB - The use of lyodura prevents the loss of hydroxyapatite crystals from the operated area during the healing period. Its protective membrane effect prevents infection and other problems during healing. Histological studies have shown that the combined use of "Ceros 80" hydroxyapatite and lyodura within 20 months resulted in the development of new, osseointegrated considerable amount of bone. This bone hydroxyapatite bond is the base for further implantological and prosthodontic procedures. PMID- 8243735 TI - [Electrochemical properties of dental alloys (II)]. AB - The studies were focused on the corrosion stability of dental alloys (containing NiCr and stainless steel), for a possible release of Nickel (Ni). The investigations were carried out in artificial saliva at 37 degrees C. The potential range, where Ni dissolution may occur, was determined by transient measurements. It has been established, that: 1. Under in vivo conditions the probability of the pitting corrosion is very low. 2. The Ni release takes place on the temporarily damaged, locally activated parts of the surfaces. PMID- 8243736 TI - [Improvement of the stability of the total lower prosthesis by vestibuloplasty]. AB - An up-to-date method of vestibuloplasty is shown by a case report. The flap raised from the cheek is suitable to cover the surface of the anterior alveolar process made free during the deepening of the lower vestibular fold. The recipient and donor areas are both within the oral cavity, thus the procedure is simple and the patients can bear it without difficulty. Due to the new anatomical situation the functional stability of complete dentures is considerably increased. PMID- 8243737 TI - [Malignant tumor developing in transplanted tissue in the oral cavity (case report)]. AB - Primary and metastatic malignant tumors appears frequently in the head and neck region. The occurrence of malignancies in tissues--transplanted to this area--are extraordinary rare. Authors report about a case, where the tumor developed in a cylindroid jump flap, used to the closure of the cleft palate. They discuss theoretical aspects, as well as the course of the disease. PMID- 8243738 TI - [Additional roentgen-anatomical examination of OP (orthopantomography) pictures]. AB - The authors examined various anatomical details which appeared in the OP pictures with varying frequency and in various recognisable forms. They sought to help to a more perfect implementation and evaluation of techniques which are considered to be relatively new in our country. PMID- 8243739 TI - [The use of iontophoresis in dental practice]. AB - The effect of hypersensitivity reducing local NaF-gel treatment with or without iontophoresis was evaluated. Air test was used for pain and pain reduction evaluation. Three consecutive treatments (0, 2, 5 weeks) and a 3 month control examination was performed on 20 and 16 patients. Additional iontophoretic treatment with Pikomed-Dention equipment was found more effective than topical NaF alone. Therapeutic use of iontophoresis could be employed in several other areas of dentistry. More attention should be paid to its use in caries prevention. PMID- 8243740 TI - [Indications for preventive removal of impacted third molars]. AB - The impacted third molars are clinically important due to the pathological alterations caused by them. It is important from orthodontic viewpoint, that during their development the anomalies combined with the constriction, or unsuitable length of the dental arch may lead to recrudescence. For this reason the preventive removal of impacted third molars is suitable at the proper stade of their development. PMID- 8243741 TI - [Basics of acupuncture in the oral cavity--oral acupuncture]. AB - In accordance with Professor Gleditsch's chinese doctrines the mouth cavity and it's surrounding areas are ensnared with energetic channels. The appropriate knowledge of meridian points is a decisive factor in making diagnoses of mouth cavity processes. There is an increasing responsibility of persons working around mouth cavity in recognising and correctly interpreting the connections between the mucous membrane processes and other remote organs and organ systems. PMID- 8243742 TI - [Quality control of dental radiography]. AB - The quality assurance of dental X-ray films can be made by a simple device prepared by Crabtree. Using this tool it is possible to check-up dental films to assure consistent radiographic quality. The radiation done of patient will be "as low as reasonably achievable". PMID- 8243743 TI - [A case of hygroma of the neck]. AB - The authors describe a case of cervical cystic hygroma in a young adult. The histological diagnosis of an operation performed two years earlier was erroneous. In this context they describe the pathology, occurrence and therapy of the disease. PMID- 8243744 TI - [Testing the reliability of face bow registration]. AB - The reliability of the Quick ear face-bow to transfer an arbitrary hinge axis to a Quick articulator was clinically compared with a Dentatus face-bow. 40 measurements were carried out on 8 patients with both instruments and were compared. The statistical analysis of the data indicated no difference between these facebow recordings. Furthermore both showed good reproducibility. This suggests that the same clinical results can be reached by these instruments. PMID- 8243745 TI - [Relation of the histological picture of biopsy material and prognosis in cases of squamous cell carcinoma of the oral cavity]. AB - Biopsy specimens from 42 oral cancer cases were graded by a histological scoring system which was initiated by Bryne et al. This system was based on both the features of the tumor cells and the relationship between tumor cells and connective tissue. The results suggested significant correlation between the histological score values and the survival, the occurrence of recidive tumors and metastatic tumors. PMID- 8243746 TI - [Dietary habits and oral hygiene in school children in the city of Debrecen]. AB - Acid products formed by bacterial fermentation of dietary carbohydrates in the dental plaque, are required for the initiation of dental caries. The aim of the present study was to analyze nutritional and oral hygienic habits of 216 schoolchildren (108 boys and 108 girls) aged of 7, 12 and 14 years. In the studied population children brushed their teeth after meals in 42.5, 54.3 and 38.1% (in the 7, 12 and 14 year age groups respectively). The additional hygienic aids (interdental cleaning: 32.5, 42.4 and 34.5%, oral rinse: 7.5, 27.2 and 23.8%) were used only by a smaller part of the studied groups. Consumption of dairy products and fruits was satisfactory. The habit of drinking carbonic acid containing soft drinks was rather high (%). Some (2.5, 10.9 and 7.1%) of the children drink it several times a day. Fluoride containing Dentocar tablets were used only in 42.5, 3.3 and 1.2% in the studied groups. PMID- 8243747 TI - "The gingiva is red around my crowns"--a differential diagnosis. PMID- 8243748 TI - The reality of dental marketing--1993. PMID- 8243750 TI - What is success? Editorial board agrees--dollars are not the measure. PMID- 8243749 TI - Videos enhance patient compliance. PMID- 8243751 TI - Maximizing your facility. PMID- 8243752 TI - Investing in your practice. PMID- 8243753 TI - PSR provides new patient-management tool. PMID- 8243754 TI - Update on computers. PMID- 8243755 TI - What is your practice really worth? PMID- 8243756 TI - Looking toward future growth. PMID- 8243757 TI - Counterclockwise crown prep. PMID- 8243758 TI - Hybridizing dentin--a new concept in treating dentinal surfaces. PMID- 8243759 TI - Current dilemmas in infection control. PMID- 8243760 TI - Periodontal regeneration with bone grafts. PMID- 8243761 TI - Effective utilization of the preperiodontal biologic provisional. PMID- 8243762 TI - Reconstruction of damaged edentulous ridges for implant placement. PMID- 8243763 TI - It's time for me to go. PMID- 8243764 TI - The components of quality dental records. PMID- 8243765 TI - Survey of dental fees. PMID- 8243766 TI - Early orthodontic treatment is important. PMID- 8243767 TI - A home away from home. PMID- 8243768 TI - Female dentists' fees higher for 35 services. PMID- 8243769 TI - Critical first step to success in dentistry. PMID- 8243770 TI - The emotional aspects of dentistry. PMID- 8243771 TI - The real costs of infection control. PMID- 8243772 TI - The laser's role in soft-tissue management. PMID- 8243773 TI - Root proximity a risk factor in periodontal disease. PMID- 8243774 TI - Current concepts in successful posterior class II direct composites. PMID- 8243775 TI - The bonded amalgam restoration. PMID- 8243776 TI - Are we becoming a profession of elitists? PMID- 8243777 TI - Take the plunge. PMID- 8243778 TI - Periodontics and dentistry's future. PMID- 8243779 TI - Putting it all together. PMID- 8243780 TI - Unconscious advertising. PMID- 8243781 TI - Gain control by giving it up. PMID- 8243782 TI - Schedule your way to improved efficiency. PMID- 8243783 TI - High-tech, but warm and inviting. PMID- 8243784 TI - Your guide to municipal bonds. PMID- 8243785 TI - Lean and serene. PMID- 8243786 TI - Success, available to most. PMID- 8243787 TI - Periodontal-flap management is improved with radiosurgery. PMID- 8243788 TI - Use and abuse of antibiotics in periodontal therapy. PMID- 8243789 TI - Successful treatment of concomitant acute myeloblastic leukaemia and adenoid cystic carcinoma of the palate. AB - The case of a 55-year old man who was affected simultaneously by acute myeloblastic leukaemia (AML) and adenoid cystic carcinoma of the palate is reported. The carcinoma was removed after induction chemotherapy for AML and the patient subjected to consolidation therapy. No reports are evident of other cases in which acute leukaemia and other malignancies were treated simultaneously, and in which the patient was cured of both. It is suggested that this case may serve as a reference for future similar cases. PMID- 8243790 TI - Dose-dependent effect of hydroxymethylglutaryl-coenzyme A reductase inhibitor on serum cholesterol with limited dietary restrictions: a case study. AB - Hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor (pravastatin sodium) can selectively inhibit cholesterol biosynthesis in the liver and may lower serum cholesterol concentrations even where there are no particular dietary restrictions. A 72-year old housewife with non-insulin-dependent diabetes mellitus complicated by hyperlipaemia type IIb, who did not follow directions for diet therapy or kinesitherapy, was administered HMG-CoA reductase inhibitor. The initial dose of 10 mg/day HMG-CoA reductase inhibitor was increased by 10 mg/day every 4 weeks to 30 mg/day, maintained at 30 mg/day for 8 weeks and then reduced gradually until discontinuation after a further 27 weeks. Test results showed the changes in low-density lipoprotein cholesterol and apoprotein B to be dose dependent. The findings represent the first clinical evidence that hypercholesterolaemia can be adequately managed by the use of HMG-CoA reductase inhibitor, even when no specific dietary restrictions are imposed, and may contribute to improvements in the quality of daily life for many patients suffering from hyperlipaemia type IIb. PMID- 8243791 TI - A parallel-group comparison of budesonide and beclomethasone dipropionate for the treatment of perennial allergic rhinitis in adults. AB - The efficacy and safety of the two intranasal glucocorticosteroids, budesonide and beclomethasone dipropionate (BDP), were compared in a single-blind study of 120 patients with perennial allergic rhinitis. Patients were randomized to receive either budesonide 200 micrograms twice daily or BDP 200 micrograms twice daily from pressurized nasal inhalers for 3 weeks. No other anti-rhinitis medication was allowed during the study. Patients recorded symptoms (blocked nose, runny nose, itchy nose, sneezing, runny eyes and sore eyes) daily on a diary card and were asked to make a global assessment of their treatment at the end of the study. Mean daily symptom scores were lower in patients receiving budesonide than in those receiving BDP for all symptoms, although treatment differences only reached statistical significance for runny nose and sneezing. After 3 weeks, 38% of patients on budesonide and 27% on BDP were totally symptom free, and 72% of patients on budesonide described the treatment as noticeably, very or totally effective compared with 58% on BDP. In conclusion, the present study showed budesonide to be more effective in controlling symptoms of perennial allergic rhinitis than BDP. PMID- 8243792 TI - A double-blind crossover study of two cyclobenzaprine regimens in primary fibromyalgia syndrome. AB - In a double-blind crossover study, the efficacy and tolerability of oral cyclobenzaprine administered in two different regimens were compared in 40 patients affected by primary fibromyalgia syndrome. The patients were randomly divided into two groups. Each group of 20 patients was treated for 15 days with either a single dose of 10 mg/day cyclobenzaprine at bedtime or 30 mg/day cyclobenzaprine in three equal doses daily. Following treatment there was a 15 day washout period before the groups were crossed over to the other treatment. Both regimens resulted in a significant decline in the number of tender points; significant improvements were also reported in the quality of sleep, anxiety, fatigue, irritable bowel syndrome and stiffness. There was no significant difference in efficacy between the two therapeutic regimens at any stage during the trial. The frequency of reported side-effects was significantly greater (P < 0.001) when patients received 30 mg/day cyclobenzaprine (26 patients, 84%) than when they received 10 mg/day (10 patients, 27%). A dose of 10 mg cyclobenzaprine at bedtime significantly improved the symptomatology of patients affected by primary fibromyalgia syndrome. The higher dose did not further reduce these symptoms but did result in a higher incidence of side-effects. PMID- 8243793 TI - Comparative efficacy of salbutamol and salmeterol in exercise-induced asthma. AB - This study compared the protection provided by salbutamol and salmeterol against exercise-induced asthma. Asthmatic patients (n = 12) with exercise-induced asthma were exercised submaximally for 6 min on a treadmill 1, 6 and 12 h after inhalation of 200 micrograms salbutamol or 50 micrograms salmeterol. Each patient also took baseline exercise 1 h after two puffs of placebo. Two days later the drugs were administered in a double-blind trial of crossover design with an interval of 48 h between the two treatments. The main parameters measured were: air flow with a Wright flowmeter and mediator concentrations (histamine, leucotriene and prostaglandin D2 measured by radioimmunoassay) in venous blood, which was withdrawn before and 4 min after each exercise period. The maximum percentage bronchoconstriction recorded following placebo was 29 +/- 4% and following salbutamol inhalation it was 4 +/- 4%, 20 +/- 13%, 27 +/- 10%, respectively, for the exercise periods performed 1, 6 and 12 h after inhalation of the drug. Following salmeterol, the corresponding figures were 3 +/- 4%, 3 +/- 3% and 11 +/- 9%. The concentrations of mediator in plasma were significantly increased after exercise. Salbutamol and salmeterol intake reduced these concentrations both when the patients were at rest and following the exercise period. This effect of both drugs on the mediators corresponded with the protection they provided against exercise-induced asthma and was maintained for 12 h after salmeterol inhalation and for 6 h after salbutamol inhalation. PMID- 8243794 TI - Non-steroidal anti-inflammatory drug-induced gastropathy: a comparative endoscopic and histopathological evaluation of the effects of tenoxicam and diclofenac. AB - A 4-week double-blind study compared the potential for 20 mg/day tenoxicam or 100 mg/day diclofenac sodium to induce gastropathy in 36 patients with joint disease and assessed the influence of gastric colonization by Helicobacter pylori. Endoscopic assessment at the end of 4 weeks indicated that the mucosa was normal in 79% of tenoxicam-treated patients and 59% of diclofenac-treated patients. Only 5% of patients in the tenoxicam group developed severe gastroduodenitis (> 11 haemorrhages or erosions) compared with 18% in the diclofenac group. Histological evaluation indicated that 58% and 47%, respectively, of tenoxicam-treated and diclofenac-treated patients retained normal mucosa after treatment. Diclofenac treatment was discontinued in two patients, due to a duodenal ulcer or severe erosive gastritis. Overall, 5/14 patients with moderate to severe colonization with Helicobacter pylori developed severe chronic active gastritis or ulceration, compared with the 1/22 patients in whom colonization was either absent or mild (P = 0.02). Tenoxicam and diclofenac did not show major differences in terms of gastrointestinal safety, although the trends favoured tenoxicam. The presence of severe colonization of the gastric mucosa with Helicobacter pylori appears to be an important factor for development of severe gastritis or ulceration. PMID- 8243795 TI - Treatment of lower respiratory tract infections with amoxycillin/clavulanic acid in adults. AB - Patients with lower respiratory tract infections [pneumonia (n = 16), bronchiectasis (n = 5) and acute exacerbations of chronic bronchitis (n = 44)] were treated daily with amoxycillin/clavulanic acid given either 1.2 g intravenously three times daily or 625 mg orally three times daily for 7-15 days. Symptoms, signs and sputum volume and colour were monitored daily. Chest X-ray, sputum culture and Gram-stain examinations were also carried out on days 1 and 5, and immediately after the end of the treatment. There was a clinical improvement, as indicated by the incidence of cough, dyspnoea and rales, and by sputum volume and colour in 90.8% of the patients. Microbiological improvement, as indicated by the complete elimination of sputum pathogens and pus cells, was achieved in the same proportion of patients (90.8%). In one patient, an adverse side-effect, diffuse exanthema, was noted. Amoxycillin/clavulanic acid possesses a high clinical and microbiological efficacy for lower respiratory tract infections. PMID- 8243796 TI - Establishment of steroidogenic granulosa cell lines expressing follicle stimulating hormone receptors. AB - Follicle stimulating hormone (FSH) plays an important role in the regulation of oogenesis, spermatogenesis and production of steroid hormones. Receptors to FSH, which are uniquely expressed in ovarian granulosa and testicular Sertoli cells, are rapidly lost in tissue culture conditions and upon cell transformation. We have succeeded, by triple transfection of primary rat granulosa cells with SV40 DNA, Ha-ras oncogene and an FSH receptor expression plasmid, to establish stable steroidogenic cell lines expressing FSH receptors. The cell lines respond to rat, ovine and bovine FSH, which stimulate progesterone production at levels comparable to primary granulosa cells obtained from preovulatory follicles. No steroidogenic response is detected upon stimulation with ovine luteinizing hormone or human chorionic gonadotropin. The steroidogenic response is accompanied by de novo appearance of adrenodoxin which serves as a marker for the mitochondrial steroidogenic enzyme system. These cells express approximately 27,000 receptors per cell with a Kd of 100-115 pM. This Kd is close to the value calculated for the native receptor. The ED50 for the steroidogenic response to ovine FSH is 200 pM, suggesting a tight coupling between receptor activation and the steroidogenic response. FSH induces pronounced morphological changes in the established cell lines, which are also characteristic of primary granulosa cells. These FSH responsive cell lines can serve as a useful model for the study of the structure and function of the FSH receptor and the effect of oncogenes on its expression. PMID- 8243797 TI - Cloning and functional characterisation of the human TRH receptor. AB - Thyrotrophin-releasing hormone (TRH) is a hypothalamic tripeptide known to act via its receptor in the anterior pituitary gland to stimulate the release of thyrotrophin (TSH) from thyrotrophs and prolactin (PRL) from lactotrophs. It is also thought to act as a neurotransmitter/neuromodulator in the central and peripheral nervous systems. We have isolated a cDNA encoding the human pituitary TRH receptor (TRH-R) protein with a predicted amino acid sequence of 398 amino acids. Comparison of the amino acid sequences of the human TRH-R with the previously published rodent TRH-Rs showed that it is similar to both the rat and mouse TRH-Rs, except that each species has variant amino acids at the carboxy (COOH) terminus. The human TRH-R belongs to the family of seven transmembrane domain, G-protein-coupled receptors, and it is believed that the COOH terminal region of this family of receptors may play an important role in receptor downregulation/internalisation, and possibly G-protein coupling. COS-1 cells expressing the human TRH-R showed high affinity receptor binding. Stimulation of these cells with TRH produced a typical phosphoinositide response and mobilisation of intracellular calcium [Ca2+]i. PMID- 8243798 TI - Competition for electron transfer between cytochromes P450scc and P45011 beta in rat adrenal mitochondria. AB - Rat adrenal mitochondria contain approximately equal levels of P450scc and P45011 beta, each reduced by NADPH through adrenodoxin reductase (ADX-reductase) and adrenodoxin (ADX). Constitutive cholesterol side-chain cleavage (SCC) can be increased over 20-fold through a combination of hormonal activation and inhibition of cholesterol metabolism in vivo prior to isolation of the mitochondria. This stimulation, which results from accumulated reactive cholesterol, does not significantly affect either the dependence of activities on the concentration of isocitrate (IC) and succinate (SU) or the ratio of maximum activities [3:1] supported by these reductants. Thus, the rate of cholesterol SCC is determined independently by electron transfer and the amount of reactive cholesterol. Hydroxylation of deoxycorticosterone (11 beta and 18 positions) required much higher levels of each reductant, indicating less effective reductant transfer to P45011 beta. Reactions at P450scc and P45011 beta, mediated by IC, are enhanced by low concentrations of various dicarboxylates anions (fumarate, SU). The actions of SU dehydrogenase inhibitors and the activity of fumarate, a poor direct reductant, suggest that higher production of NADPH results from malate-enhanced uptake of isocitrate. Only synergistic combinations of reductants are sufficient to sustain maximum rates of 11-deoxycorticosterone (DOC) metabolism, whereas IC is fully effective for P450scc. Increased reaction at P450scc (cholesterol loading or addition of 20 alpha-hydroxycholesterol) decreased simultaneous DOC metabolism at P45011 beta in inverse proportion to the estimated intramitochondrial generation of NADPH (1 mM or 10 mM SU > 1 mM IC > 10 mM IC). These decreases were reversed by inhibition of P450scc. Crossover inhibition caused by maximum DOC metabolism was less pronounced. EGTA/albumin treatment, which enhanced activities at both P450scc and P45011 beta, presumably via increased NADPH, diminished this cross-competition. The differential dependence on reductants and the characteristics of crossover competition are consistent with a roughly three-fold more favorable partitioning of electron transfer to P450scc, possibly caused by preferential interaction of reduced adrenodoxin with P450scc. PMID- 8243799 TI - Promoter independent down-regulation of the firefly luciferase gene by T3 and T3 receptor in CV1 cells. AB - We report that the activity of the firefly luciferase (LUC) reporter gene is down regulated by T3 and T3 receptor (TR) in the CV1 mammalian cell line, which is widely used for studies of TR action. Repression was highly reproducible, T3 and TR dependent, promoter independent, and observed regardless of whether an internal control for transfection efficiency was used. Cotransfections with normal and mutant TRs indicate that the negative T3 response is mediated by sequences within the LUC gene coding region, and is not due to the interaction of TR with a limiting transcription factor. Negative regulation of the LUC reporter was overcome by a strong, cis-linked T3 response element (TRE), but continued in the presence of a TRE of moderate strength. The results described here demonstrate that conclusions drawn from studies of TRE structure and activity performed using the LUC reporter in CV1 cells should be interpreted with caution. PMID- 8243800 TI - Biosynthesis of ecdysteroid hormones by crustacean Y-organs: conversion of cholesterol to 7-dehydrocholesterol is suppressed by a steroid 5 alpha-reductase inhibitor. AB - A pair of glands (Y-organs) in crustaceans synthesize and secrete ecdysteroid hormones; the obligate precursor for synthesis is circulating cholesterol. Ecdysteroid output by the Y-organs is regulated negatively by an eyestalk neurosecretory peptide, molt-inhibiting hormone (MIH). The question was addressed, does MIH suppress ecdysteroid synthesis by decreasing cholesterol supply (uptake) or its utilization or both? Experiments were conducted with Y organs in vitro from the crab, Menippe mercenaria, in the presence of labeled cholesterol, with or without the steroid 5 alpha-reductase inhibitor, L-645390 (Merck). Other experiments superimposed the presence or absence of eyestalk extract containing MIH activity. L-645390 greatly depressed incorporation of cholesterol into an early intermediate, 7-dehydrocholesterol and into secreted ecdysteroids. At the same time, cholesterol accumulated in the Y-organs, to levels significantly higher than in untreated controls. MIH alone depressed both cholesterol uptake and incorporation. MIH together with L-645390 produced the greatest depression of cholesterol incorporation while also preventing the cholesterol accumulation seen with L-645390 alone. These results indicate that cholesterol uptake and its metabolic utilization in Y-organs are separable events representing separate sites of regulation by MIH. PMID- 8243801 TI - Localization of the glucocorticoid receptor in testis and accessory sexual organs of male rat. AB - Localization of glucocorticoid receptor-like immunoreactivity (GR-LI) was studied in adult rat testis, epididymis, ejaculatory duct, seminal vesicle and prostate by light and electron microscopic immunocytochemistry. In the interstitium of the testis GR-LI was seen in the nuclei of Leydig cells, macrophages, fibroblasts, smooth muscle cells and endothelial cells of blood vessels. Furthermore, GR-LI was observed in zygotene and early pachytene primary spermatocytes of some seminiferous tubules during stages XIII-XIV and I-III of the spermatogenic cycle. Other spermatogenic cells and Sertoli cells were devoid of staining. GR-LI was also found in peritubular myoid cells, fibroblasts and basal cells of the epididymis, vas deferens and prostate. Localization of GR-LI in primary spermatocytes and Leydig cells suggests that glucocorticoids directly affect spermato- and steroidogenesis in the testis. The absence of GR-LI from functional, stromal cells of the male accessory sexual organs suggests that they are not targets for glucocorticoid hormones. PMID- 8243802 TI - Dopamine D2 receptor mRNA in the pituitary during the oestrous cycle, pregnancy and lactation in the rat. AB - We have used the technique of quantitative in situ hybridization in order to study the changes in the levels of expression of D2 receptor mRNA in the anterior pituitary gland of female rats at different stages of the reproductive cycle. Plasma prolactin levels in the same animals were determined by radioimmunoassay. Rats in the prooestrous, oestrous, dioestrous 1, dioestrous 2 phases of the oestrous cycle and in pregnant, lactating, ovariectomised and ovariectomised animals treated with diethylstilbestrol (DES) have been examined. Our results show that expression of D2 receptor mRNA in the anterior pituitary gland varies during the oestrous cycle, with the lowest expression measured during oestrus. Expression levels increased during dioestrus 1, reaching the highest values in dioestrus 2 and declining again in prooestrus. Expression of D2 receptor mRNA was reduced during pregnancy when compared to lactating animals. In ovariectomised animals, the level of D2 receptor mRNA was similar to that observed in intact animals during oestrus. Ovariectomised animals treated with DES showed dramatically increased prolactin levels, while D2 receptor mRNA remained low. Prolactin secretion might be controlled not only by variations in the release and plasma concentrations of dopamine itself, but also by modulation of D2 receptor expression in pituitary cells. Our results suggest that the variations in D2 receptor density in the anterior pituitary cells at different physiological states are, at least to some extent, regulated at the level of gene expression. PMID- 8243803 TI - Acute cholecystitis potentiates bradykinin stimulated fibroblast prostanoid release in the rabbit. AB - Gallbladder explants from control rabbits and rabbits subjected to bile duct ligation (BDL) for 24 and 72 h (cholecystitis model) were placed in cell culture to determine the source for increased gallbladder prostanoid synthesis during cholecystitis. Cultures from control and 24 h BDL gallbladders grew spindle shaped fibroblasts which did not exhibit increased prostanoid synthesis. 72 h BDL gallbladder cell cultures grew large polygonal shaped cells which appeared to be 'stimulated fibroblasts' by light and electron microscopy and were associated with increased basal and bradykinin stimulated 6-keto-PGF1 alpha release and increased content of prostacyclin synthase when measured by enzyme immunoassay and protein immunoblot analysis respectively. Use of bradykinin antagonists showed that the bradykinin BK2 subtype receptor was the most prominent in the 72 h BDL cell cultures. The 'stimulated fibroblasts' were the source of bradykinin stimulated gallbladder 6-keto-PGF1 alpha synthesis in the inflamed rabbit gallbladder which was mediated by the bradykinin B2 subtype receptor. PMID- 8243804 TI - Human chorionic gonadotropin induces interleukin-1 gene expression in rat Leydig cells in vivo. AB - Luteinizing hormone (LH)/human chorionic gonadotropin (hCG) causes inflammatory type responses in the testis. In the present study, we evaluated the effects of hCG on Leydig cell interleukin-1 (IL-1) gene expression. Using monoclonal antibody (ED2) staining for macrophages, our Leydig cell preparations had no significant contamination with macrophages. When purified Leydig cells from normal rats were cultured for 24 h and then treated with IL-1 beta (1-100 ng/ml) for 6 h, IL-1 beta induced dose-dependent increases in IL-1 beta mRNA levels. IL 1 beta also induced IL-1 alpha mRNA accumulation; however, the level of IL-1 alpha mRNA was much lower than that of IL-1 beta mRNA. When rats were treated with either saline or hCG (5 units i.p.), hCG markedly induced IL-1 beta mRNA accumulation in purified Leydig cells at 6 h which persisted for over 24 h. However, hCG had no direct effect on purified Leydig cell or crude interstitial cell IL-1 mRNA levels. Our results suggest that inflammatory effects of hCG in vivo may be mediated by increased IL-1 gene expression in Leydig cells. PMID- 8243805 TI - Estrogen receptor binds to the salmon GnRH gene in a region with long palindromic sequences. AB - Footprinting and gel shift assays demonstrated that the human estrogen receptor (hER) specifically binds to two estrogen response element (ERE)-like motifs in the gonadotropin releasing hormone (GnRH) gene promoter region of Atlantic salmon (Salmo salar). The two ER binding sites are situated approximately 1.5 kb upstream of the transcriptional start site of the GnRH gene and are localized 49 bp from each other. Each ERE-like motif is composed of two palindromic ERE half sites interspaced by 8 and 9 nucleotides, respectively. The salmon GnRH gene promoter region contains an almost perfect 426-bp-long palindromic sequence that might form a cruciform structure. PMID- 8243806 TI - Differential expression of keratinocyte growth factor and its receptor in the human uterus. AB - The expression of mRNAs for keratinocyte growth factor (KGF) (also called FGF-7) and its receptor was evaluated in normal human endometrium and myometrium as well as in myoma and in endometrial adenocarcinoma cell lines using reverse transcriptase polymerase chain reaction. Both KGF and its receptor mRNA are expressed in the human endometrium throughout the menstrual cycle, whereas fibroblast growth factor receptor 2 (FGFR-2) mRNA expression is low in this tissue. In endometrial stromal cell enriched preparations KGF mRNA dominates with little expression of KGF receptor (KGFR) and FGFR-2, whereas in the epithelial cell-enriched fraction the KGFR mRNA dominates. Human myometrium and myoma express mRNA for KGF, but not for KGFR. FGFR-2 is expressed in both myometrial and myoma tissues. None of the five endometrial adenocarcinoma cell lines studied expressed KGF mRNA, whereas all cell lines expressed mRNA for either KGFR or FGFR 2 or for both receptors. The results show a selective expression of KGFR and the closely related FGFR-2 in the human uterus with the former being expressed in the endometrium and the latter predominantly in the adjacent myometrium. In the endometrial tissue, selective expression of KGF in stromal cells and KGFR in epithelial cells supports the paracrine function of KGF in epithelial tissue. PMID- 8243807 TI - 1,25(OH)2-vitamin D3 mediated changes in mRNA for c-myc and 1,25(OH)2D3 receptor in HL-60 cells and related subclones. AB - HL-60 cells have been shown to differentiate into monocyte-like cells as a consequence of the interaction of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) with a specific intracellular receptor (VDR). Of c-myc it has been reported that it plays a role in cell proliferation and differentiation. We have investigated the expression of VDR and c-myc mRNA of three subclones of HL-60 cells (N, R and MR) after exposure to 1,25(OH)2D3. The N cell is a normal clone in which 1,25(OH)2D3 inhibited cell proliferation and also induced cell differentiation. The R cell is a resistant clone whose proliferation and differentiation are not affected by 1,25(OH)2D3. The MR cell is a mixed response type clone whose proliferation was not inhibited by 1,25(OH)2D3, while its differentiation was actually induced by 1,25(OH)2D3. The VDR mRNA expression of the N and MR cells reached a peak at 2 h (2.3- and 2.6-fold induction, respectively) and returned to the control level after 24 h treatment of 1,25(OH)2D3. The c-myc mRNA expression was significantly inhibited in the 1,25(OH)2D3-induced N cells, but not in the MR cells. In contrast, 1,25(OH)2D3 did not induce any changes of the VDR and c-myc mRNA levels in the R cells. In separate experiments, the level of the VDR in the three clones was determined via Scatchard analysis; the VDR of the N, MR and R cells were found to be 4500/cell, 3570/cell and less than 600/cell, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243808 TI - Growth hormone-receptor messenger RNA in the rat ovary: regulation and localization. AB - The results of several reports indicate that GH can modulate ovarian function. In the present study, the expression of the growth hormone-receptor (GH-R) mRNA was studied in the rat ovary using an RNA probe corresponding to a part of the extracellular domain of the GH-R. The probe hybridized to two major transcripts with estimated sizes of 4.0 kb and 1.2 kb in RNA extracted from liver and ovary. Recently, these transcripts have been shown to encode the GH-R and the GH-binding protein (GH-BP). The ontogeny of the GH-R/GH-BP mRNA expression was studied using Northern blot analysis and a solution hybridization RNase protection assay. In the liver GH-R/GH-BP mRNA levels increased with age, while in the ovary, the levels decreased between 1 and 5 weeks of age. Hypophysectomy caused a decrease in GH-R/GH-BP gene expression in the ovary, an effect which could be partly reversed with a single injection of GH (2 mg/kg). No significant changes in the ovarian concentration of GH-R/GH-BP transcripts were seen during the estrus cycle. Using in situ hybridization GH-R/GH-BP transcripts were found to be most abundant in follicles. Northern blot analysis of RNA extracted from isolated granulosa cells and corpora lutea showed that both these compartments contained GH-R and GH-BP mRNA, although more abundant in granulosa cells. Immunoreactive GH R was detected in granulosa cells of healthy follicles, corpus luteum, and in the germinal epithelium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243809 TI - Thyrotropin-releasing hormone and gonadotropin-releasing hormone-associated peptide modulation of [Ca2+]i in human lactotrophs. AB - The effect of thyrotropin-releasing hormone (TRH) and gonadotropin-releasing hormone-associated peptide (GAP) was studied on both secretion and intracellular free Ca2+ concentrations ([Ca2+]i) in human pituitary cells cultured from prolactin (PRL)-secreting tumors. Secretion was measured during a 30-min incubation period and we used a microspectrofluorimetric method in individual cells and indo-1 as the fluorescent probe. TRH (10(-8) M) significantly increased PRL release in five out of the six cell populations. In these five cases, more than 68% of individual cells responded to TRH by an increase in [Ca2+]i. No significant increase in PRL secretion was found in another culture in which TRH increased [Ca2+]i in only 37% of the cells. The effect of GAP (10(-7) M) was studied in five cell populations. In three of them, a decrease of 20% to 51% of the PRL basal secretory rate was observed under GAP. GAP inhibited [Ca2+]i in respectively 59%, 46% and 94% of the cells from these cultures. The inhibitory effect of GAP was blocked by a pertussis toxin (PT) pretreatment which demonstrates the involvement of a PT-sensitive G-protein in GAP action. In two other cultures, GAP did not significantly alter PRL secretion or individual cell [Ca2+]i. These observations suggest that GAP might play a role in the control of PRL secretion in the human. PMID- 8243810 TI - Differential involvement of phospholipase A2 in phorbol ester-induced luteinizing hormone and growth hormone release from rat anterior pituitary tissue. AB - The protein kinase C (PKC) activator, phorbol 12,13-dibutyrate (PDBu) induced the release of both luteinizing hormone (LH) and growth hormone (GH) from proestrous rat anterior pituitary pieces in vitro. Phorbol 12,13-dibutyrate-induced LH, but not GH release was readily inhibited by the phospholipase A2 (PLA2) inhibitors, quinacrine, aristolochic acid, ONO-RS-082 and chloracysine. Furthermore, PDBu induced release of [3H]arachidonic acid ([3H]AA) from pre-labelled anterior pituitary tissue that was prevented in the presence of quinacrine, aristolochic acid and ONO-RS-082 but not the diglyceride lipase inhibitor RHC 80267. The effect of PDBu was completely inhibited by staurosporine and the selective PKC inhibitor Ro 31-8220 but only partially by low concentrations of H7; consistent with the involvement of both H7-sensitive and H7-resistant forms of PKC in the activation of PLA2 by PDBu. The protein synthesis inhibitor cycloheximide inhibited the release of both [3H]AA and LH that had been induced by PDBu, whereas LH release induced by the PLA2 activator mellitin was cycloheximide insensitive. These results suggest that PKC activators may induce LH but not GH release from anterior pituitary tissue by a mechanism involving activation of a PLA2, brought about by a process which is reliant on protein synthesis. PMID- 8243811 TI - In the thyroid cells proliferation, differentiated and metabolic functions are under the control of different steps of the cyclic AMP cascade. AB - In the course of studies to elucidate the complex network of interactions controlling FRTL5 cell proliferation, thyroid stimulating hormone (TSH) independent mutants (M cells), have been obtained from FRTL5 cells by chemical mutagenesis. In the present studies, the role of TSH on the proliferation and on differentiated and metabolic functions in these mutant cells have been investigated and compared to their response to insulin-like growth factor I (IGF I). The addition of IGF-I to M cells leads to normal stimulation of DNA synthesis. However, inspite of the fact that mutant cells display normal TSH receptors, TSH is unable to stimulate the proliferation of the M cells. Nevertheless, TSH is able to increase intracellular levels of cAMP leading to regulation of TSH function in the M cells. On the other hand, TSH does not influence iodide transport and actin filaments depolimerization in these cells. However, aminoacid transport, stimulated in wild-type FRTL5 cells by both TSH and IGFs, is under the control of IGFs but not of TSH in the mutant cells. Neither TSH or IGF-I modified the expression of c-fos proto-oncogene in the M cells, probably because of high constitutive expression. These data suggest that a crucial signalling step(s) required for TSH induced mitogenesis is impaired in the M cells, and that this signalling step is not required for IGF-I induced mitogenesis. PMID- 8243812 TI - The sequence of the 5'-end of the rat CYP17 gene, the transcription initiation site and a comparison with the homologous genes of other species. AB - A clone bearing the rat CYP17 gene has been isolated and the sequence determined for 417 nucleotides 5' to the translation start site. The sequence shows one difference, a G-C inversion, with that published by Zhang et al. (1992). A transcription initiation site has been located at position -41 relative to the translation start site; this confirms the location published by Nason et al. (1992) with the caveat of the G-C inversion, so that the transcription start site becomes G instead of C. Overall percent similarities of this sequence with those of bovine, human, pig and mouse show values of 58, 64, 65, and 82, respectively. Similar comparisons done for the cAMP-responsive sequences, CRS-1 and CRS-2 (Lund et al., 1990), show 42, 55, 47 and 95% similarities for CRS-1, and 56, 77, 71 and 90% for CRS-2. These comparisons suggest that the highly conserved rat and mouse gene sequences corresponding to the bovine CRS-1 and CRS-2 may be important for either negative regulation of this gene in the adrenal or positive regulation in the gonads of these two species. PMID- 8243813 TI - Hypoglycemia begets hypoglycemia in IDDM. PMID- 8243814 TI - Inhibition of lipolysis decreases lipid oxidation and gluconeogenesis from lactate but not fasting hyperglycemia or total hepatic glucose production in NIDDM. AB - We determined whether overnight inhibition of lipolysis by a long-acting nicotinic acid derivative (acipimox) decreases gluconeogenesis from lactate in NIDDM patients. For this purpose, 250 mg of acipimox or placebo was administered in a double-blind crossover study at 2400, 0400, and 0800 to 8 NIDDM patients (54 +/- 4 yr of age, body mass index 29.5 +/- 1.3 kg/m2, fasting plasma glucose 11 +/ 1 mM). The next morning, total hepatic glucose production (glucose Ra) and gluconeogenesis from lactate were determined using primed, continuous infusions of [3-3H]glucose and [U-14C]acetate. Glucose and lipid oxidation rates were measured using indirect calorimetry. Mean overnight serum free fatty acid concentrations averaged 242 +/- 8 microM after acipimox and 721 +/- 30 microM after placebo (P < 0.001). Inhibition of lipolysis decreased lipid oxidation from 33 +/- 3 to 22 +/- 2 J.kg-1 x min-1 (P < 0.001) and increased carbohydrate oxidation from 15 +/- 3 to 23 +/- 2 mumol.kg-1.min-1 (P < 0.005). Gluconeogenesis from lactate decreased by approximately 40%, from 6.2 +/- 0.6 to 3.8 +/- 0.5 mumol.kg-1 x min-1 (P < 0.005); lactate oxidation increased from 5.6 +/- 0.8 to 7.9 +/- 1.1 mumol.kg-1 x min-1 (P < 0.005), with no change in plasma lactate concentrations or total lactate Rd. Fasting plasma glucose concentrations were comparable at 2400 (10.0 +/- 1.1 vs. 10.6 +/- 1.3 mM, acipimox vs. placebo) and between 0900 and 1000 (10.6 +/- 1.3 and 11.3 +/- 1.3 mM, respectively). Also, total glucose production rates remained unchanged (14.0 +/- 1.2 vs. 14.9 +/- 1.3 mol.kg-1 x min-1, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243815 TI - Intensive insulin therapy and weight gain in IDDM. AB - Intensive insulin therapy is frequently complicated by excessive weight gain. The purpose of this study was to determine the cause and composition of this weight gain. Therefore, changes in body composition, energy expenditure, glycosuria, and substrate kinetics were evaluated in patients with IDDM who transferred from conventional insulin therapy to intensive insulin therapy. Six adult patients with IDDM were studied on conventional insulin therapy and after 2 mo of intensive insulin therapy while maintaining constant caloric intake and were compared with a group of 6 matched nondiabetic volunteers. Body composition was determined by underwater weighing. Energy expenditure was measured during 24-h stays in a whole-room calorimeter. Whole-body turnover rates of glucose, glycerol, palmitate, and leucine were determined by isotope dilution methods. Intensive insulin therapy lowered the mean daily blood glucose concentration and HbA1 (14.8 +/- 1.6 to 7.7 +/- 0.6 mM and 12.9 +/- 0.9 to 9.6 +/- 0.6%, both P < 0.01) and almost eliminated glycosuria (428 +/- 116 to 39 +/- 22 mmol/day, P < 0.05). Body weight increased 2.6 +/- 0.8 kg with intensive insulin therapy (P < 0.05) as a result of an increase in fat mass (2.4 +/- 0.8 kg, P < 0.05). Daily energy expenditure decreased 5% (118 +/- 32 kcal/day) with intensive insulin therapy (P < 0.05). The rates of glucose, glycerol, free fatty acid, and leucine turnover, triglyceride/free fatty acid cycling, and nonoxidative glucose and protein disposal were reduced in the diabetic volunteers during intensive insulin therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243816 TI - Localization of a type I diabetes susceptibility locus to the variable tandem repeat region flanking the insulin gene. AB - A susceptibility gene for type I diabetes is present on chromosome 11p15.5, but its location, identity, and mechanism of action are unknown. We have sequenced 14 kilobases of DNA flanking the human insulin gene and found new DNA polymorphisms and determined their frequencies in the general population and in families of type I diabetic subjects. A DNA polymorphism located 3123 base pairs downstream from the initiation site of transcription of the insulin gene, when present in the homozygous state, provides a relative risk for type I diabetes of 5.2 (P = 0.006). However, this DNA polymorphism as well as other diabetes-associated 3' markers are in linkage-disequilibrium with the actual susceptibility region, because these polymorphisms are found on haplotypes both positively and negatively associated with type I diabetes susceptibility. Nucleotide sequence analysis of the variable tandem repeat region flanking the 5' end of the insulin gene shows variable tandem repeat elements associated with these haplotypes to differ greatly in composition, i.e., an ATAGGGGTGTGGGG repeat element is absent on a haplotype associated with type I diabetes susceptibility, but is found in 6 10 copies on two haplotypes negatively associated with the disease. These findings suggest that the type I diabetes susceptibility locus on chromosome 11p15.5 is probably located in the 5' variable tandem repeat region rather than in the 3' region of the insulin gene. PMID- 8243817 TI - A second pathway for regeneration of adult exocrine and endocrine pancreas. A possible recapitulation of embryonic development. AB - Substantial regeneration of both the endocrine and exocrine pancreas occurs after a 90% partial pancreatectomy in the young adult rat. We have reported previously that replication of preexisting islet and exocrine cells is enhanced 3- to 4 fold. Here, we report a second pathway of regeneration, that of proliferation and differentiation of precursor cells in the ductal epithelium. As shown with in vivo pulse labeling using 5-bromo-2'-deoxyuridine, an expansion of the ductal epithelium occurs. Proliferation is seen first in the common pancreatic duct and sequentially in smaller ducts of the ductal tree as focal areas of proliferation small ductules form. By 60 h after pancreatectomy, only these focal areas show heavy 5-bromo-2'-deoxyuridine staining. These proliferating ductules comprise 12.8% of the pancreatic volume at 3 days after pancreatectomy but are uncommon at 7 days after pancreatectomy. Coincident with the appearance and disappearance of these regions was a 3.5-fold increased growth of the pancreatic remnant compared with its equivalent of sham animals. These small ductules differentiate into new pancreatic islets and exocrine tissue, forming new lobules of pancreas that are indistinguishable from the preexisting ones. This second pathway of rapid regeneration recapitulates embryonic development in its pattern of ductal proliferation and subsequent differentiation. Furthermore, these studies provide evidence of the presence of precursor/stem cells in the adult pancreas. PMID- 8243818 TI - Antioxidant MDL 29,311 prevents diabetes in nonobese diabetic and multiple low dose STZ-injected mice. AB - Recent investigations suggest a role for antioxidants in preventing IDDM. MDL 29,311 (4,4'-[methylenebis(thio)]bis](1,1- dimethylethyl)]-phenol) is an analogue of the antioxidant probucol. Administered as a 1% dietary admixture to female nonobese diabetic mice from 4 to 24 wk of age, MDL reduced the prevalence of diabetes from 49 to 4% at 24 wk of age (n = 50-61/group). Discontinuation of treatment at 24 wk of age did not result in a rapid onset of diabetes. Probucol (1%) did not prevent diabetes. Initiating MDL treatment at 4 or 8 wk of age was more effective (19 and 17%, respectively, compared with 60% in control mice) than initiating treatment at 12 wk of age (30% diabetic; n = 28-35/group). A lower dose of MDL (0.1%), started at 4 wk of age, decreased the prevalence of diabetes to 36%. Histopathology indicated that MDL did not prevent insulitis. MDL (0.1%) also was evaluated in combination with immunosuppressants. Compared with control mice (65% diabetic), the combination of MDL and deflazacort was more effective (21% diabetic) than either agent alone (39% diabetic for MDL and 59% diabetic for deflazacort), whereas the effectiveness of MDL, cyclosporin, and MDL plus cyclosporin was similar (39, 38, and 34% diabetic, respectively). In another model of IDDM, the multiple-low-dose streptozocin-injected mouse, MDL (1%) also reduced the prevalence of diabetes when administered beginning 8 wk before streptozocin (55% diabetic vs. 100% of control mice; n = 20-25/group). Probucol (1%) was ineffective. MDL appears effective in preventing the onset of disease in two mouse models of IDDM. PMID- 8243819 TI - Impaired regulation of hepatic fructose-1,6-bisphosphatase in the New Zealand obese mouse model of NIDDM. AB - The New Zealand obese mouse, a model of NIDDM, is characterized by hyperglycemia, hyperinsulinemia, and hepatic and peripheral insulin resistance. The aim of this study was to investigate the biochemical basis of hepatic insulin resistance in NZO mice. Glycolytic and gluconeogenic enzyme activities were measured in fed and overnight fasted 19- to 20-wk-old NZO and control New Zealand chocolate mice. The NZO mice were twice as heavy as the NZC mice. The activity of the glycolytic enzymes glucokinase and pyruvate kinase was higher, whereas that of the gluconeogenic enzymes PEPCK and glucose-6-phosphatase was lower in fed and fasted NZO mice. These enzyme changes are consistent with a normal response to the hyperinsulinemia in NZO mice. In contrast, the activity of the third regulated gluconeogenic enzyme, fructose-1,6-bisphosphatase, was similar in fed and fasted NZO and NZC mice despite the higher insulin and glucose levels in the NZO mouse. This enzyme is primarily regulated by the powerful inhibitor fructose-2,6 bisphosphate. The levels of this metabolite were measured and found to be increased in both the fed and fasted states in the NZO mouse, suggesting that the activity of the bifunctional enzyme that regulates the level of inhibitor (6 phosphofructo-2-kinase/fructose-2,6- bisphosphatase) is normally regulated in the NZO mouse. We conclude that most insulin-responsive gluconeogenic and glycolytic enzymes are normally regulated in the NZO mouse, but an abnormality in the regulation of fructose-1,6-bisphosphatase may contribute to the increase hepatic glucose production in these mice. PMID- 8243820 TI - Mechanisms for D-glucose inhibition of myo-inositol influx into rat lens. AB - Myo-Inositol depletion as a result of hyperglycemia is considered one of the leading contributors to chronic diabetic complications. We investigated the possible mechanisms through which elevated extracellular glucose levels affect the loss of intracellular myo-inositol in rat lens. Short-term incubation (up to 4 h) in solutions with elevated glucose concentrations revealed a concentration dependent inhibition of myo-inositol influx. This inhibition was caused by both an increase of the transport coefficient and a decrease of maximal flux and thus was a mixed competitive and noncompetitive inhibition. If polyol accumulation was prevented with sorbinil, an aldose reductase inhibitor, the inhibition of myo inositol influx was partially reduced. The remaining inhibition was the result of an increased transport coefficient without a change in maximal flux and therefore represents a strictly competitive inhibition. A similar competitive inhibition was observed with the nonmetabolizable glucose analogue L-glucose, which cannot be converted to polyol. Longer exposure (16 h) to solutions with high glucose concentrations resulted in an inhibition that correlated with high lens polyol levels. This inhibition persisted after the lenses were returned to solutions with normal glucose concentrations and was the result of a decrease of maximal flux without a significant change in transport coefficient, a strictly noncompetitive inhibition. The noncompetitive inhibition associated with polyol accumulation and the competitive inhibition due to extracellular glucose were additive. Lens myo-inositol depletion after exposure to elevated glucose concentrations thus resulted from a competitive inhibition caused by the interaction of extracellular glucose with the myo-inositol carrier and a noncompetitive inhibition associated with polyol accumulation. PMID- 8243821 TI - Relationship between the circadian rhythms of blood pressure and sympathovagal balance in diabetic autonomic neuropathy. AB - In diabetic autonomic neuropathy, abnormal circadian patterns of blood pressure and sympathovagal balance with reduced fall of blood pressure and prevalence of sympathetic activity during the night have been described. To correlate the abnormalities of blood pressure to those of sympathovagal balance, we simultaneously performed 24-h noninvasive monitoring of blood pressure and ECG in 25 diabetic patients (45.6 +/- 13.6 yr of age with a 17.6 +/- 9.1 yr duration of diabetes) with various degrees of cardiovascular reflex impairment. Autoregressive power spectrum analysis of RR interval variability was applied to 24-h ECG recordings to obtain for day and night periods the mean power of low- (0.03-0.15 Hz) and high-frequency (0.18-0.40 Hz) components, which are relative markers of sympathetic and vagal activity, respectively, and their ratio (low frequency/high frequency), assumed as index of sympathovagal balance. Diabetic patients showed a lower percentage of day-night change in systolic blood pressure (9 +/- 5.48 vs. 11.6 +/- 4.78%, P < 0.037), a lower day low frequency (5.9 +/- 0.81 vs. 6.62 +/- 0.73 In-ms2, P < 0.001), a lower night high frequency (6.06 +/- 0.71 vs. 6.52 +/- 0.85 In-ms2, P < 0.05), a lower day low frequency:high frequency ratio (1.82 +/- 1.77 vs. 3.05 +/- 1.82, P < 0.01), and a lower percentage of day-night change in low-frequency:high frequency ratio (-13.4 +/- 109.9 vs. 28.7 +/- 29.7%, P < 0.05), when compared with control subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243822 TI - Sulfonylurea-binding sites and ATP-sensitive K+ channels in alpha-TC glucagonoma and beta-TC insulinoma cells. AB - alpha-Cells secrete glucagon in a fuel-dependent fashion. We tested the hypothesis that alpha-cells contain sulfonylurea- and ATP-sensitive K+ channels. We studied two clonal lines of alpha-TC cells (simian virus 40 T-antigen induced glucagonoma cells) and for reference purposes, similarly transformed beta-TC insulinoma cells. alpha-TC cells each contained approximately 3000 high-affinity binding sites for the sulfonylurea [3H]glyburide. Whole-cell ATP- and tolbutamide sensitive K+ currents of alpha-TC and beta-TC cells, relative to cell surface area, were comparable. In cell-attached membrane patches of alpha-TC cells, two types of K+ channels were observed. They had slope conductances of approximately 63 and 33 pS when the electrode contained 151 mM K+. Tolbutamide and diazoxide decreased and enhanced, respectively, the open probability of these channels. The membrane of alpha-TC cells depolarized periodically. This electrical activity was inhibited by diazoxide. A physiological mixture of amino acids enhanced glucagon release, and high glucose partially inhibited this release. Tolbutamide also enhanced glucagon release, whereas diazoxide inhibited it. Thus, alpha-TC glucagonoma cells contain ATP-sensitive K+ channels that regulate glucagon release, yet allow inhibition of hormone release by glucose. PMID- 8243823 TI - Multiple defects in the adipocyte glucose transport system cause cellular insulin resistance in gestational diabetes. Heterogeneity in the number and a novel abnormality in subcellular localization of GLUT4 glucose transporters. AB - Mechanisms causing cellular insulin resistance in gestational diabetes mellitus are not known. We, therefore, studied isolated omental adipocytes obtained during elective cesarean sections in nondiabetic (control) and GDM gravidas. Cellular insulin resistance was attributed to impaired stimulation of glucose transport; compared with control subjects, basal and maximally insulin-stimulated transport rates (per surface area) were reduced 38 and 60% in GDM patients, respectively. To determine underlying mechanisms, we assessed the number, subcellular distribution, and translocation of GLUT4, the predominant insulin-responsive glucose transporter isoform. The cellular content of GLUT4 was decreased by 44% in GDM patients as assessed by immunoblot analysis of total postnuclear membranes. However, GDM patients segregated into two subgroups; half expected profound (76%) cellular depletion of GLUT4 and half had GLUT4 levels in the normal range. Cellular GLUT4 was negatively correlated with adipocyte size in the control subjects and GDM patients with normal GLUT4 (r = 0.60), but fell way below this continuum in GDM patients with low GLUT4, indicating that heterogeneity was not caused by differences in obesity. All GDM. distribution. In basal cells, increased amounts of GLUT4 were detected in membranes fractionating with (such that the plasma membrane GLUT4 level in GDM (such that the plasma membrane GLUT4 level in GDM patients was equal to that observed in insulin stimulated cells from control subjects). Furthermore, insulin stimulation induced translocation of GLUT4 from low-density microsomes to plasma membranes in control subjects but did not alter subcellular distribution in GDM patients. In other experiments, cellular content of GLUT1 was normal in GDM patients, and GLUT1 did not undergo insulin-mediated recruitment to plasma membranes in either control subjects or GDM patients. A faint signal was detected for GLUT3 only in low density microsomes and only with one of two different antibodies. In GDM, we conclude that insulin resistance in adipocytes involves impaired stimulation of glucose transport and arises from a heterogeneity of defects intrinsic to the glucose transport effector system. GLUT4 content in adipocytes is profoundly depleted in approximately 50% of GDM patients, whereas all patients are found to exhibit a novel abnormality in GLUT4 subcellular distribution. This latter defect is characterized by accumulation of GLUT4 in membranes cofractionating with plasma membranes and high-density microsomes in basal cells and absence of translocation in response to insulin. The data suggest that abnormalities in cellular traffic or targeting relegate GLUT4 to a membrane compartment from which insulin cannot recruit transporters to the cell surface and have important implications regarding skeletal muscle insulin resistance in GDM and NIDDM. PMID- 8243824 TI - Early introduction of dairy products associated with increased risk of IDDM in Finnish children. The Childhood in Diabetes in Finland Study Group. AB - Associations between infant-feeding patterns and risk of IDDM were investigated in a nationwide Finnish case-control study of 690 IDDM children < 15 yr of age. Each child was matched by date of birth and sex to a randomly selected population based control child. Univariate analysis revealed that the risk of IDDM was increased by approximately 1.5 in children for whom breast-feeding was terminated at < 2 mo of age, doubled in those who were exclusively breast-fed for < 2 mo, and doubled in those who were introduced to dairy products at < 2 mo of age. In further multivariate analyses of these factors, it was found that introduction of dairy products at an early age was the most important risk factor, and the observed univariate effects of duration of breast-feeding variables were explained by their correlation with this factor. This is the first observational study to show that early introduction of dairy products is independently associated with an increased risk of IDDM. Adjustment for mother's education and age, child's birth order, or birth weight did not affect the results. PMID- 8243825 TI - Mechanism of awareness of hypoglycemia. Perception of neurogenic (predominantly cholinergic) rather than neuroglycopenic symptoms. AB - We sought 1) to determine which symptoms of hypoglycemia are reproducible, 2) to pharmacologically distinguish neurogenic (autonomic) from neuroglycopenic symptoms, and 3) to test the hypothesis that awareness of hypoglycemia is the result of perception of neurogenic rather than neuroglycopenic symptoms. Awareness of hypoglycemia and 19 symptoms were quantitated in 10 normal, young adults, each studied on four occasions in random sequence, during 1) clamped euglycemia (approximately 5 mM), 2) clamped hypoglycemia (approximately 2.5 mM), 3) clamped hypoglycemia with combined alpha- and beta-adrenergic blockade (phentolamine and propranolol), and 4) clamped hypoglycemia with pan-autonomic blockade (phentolamine, propranolol and atropine). Significant (ANOVA, P < 0.001) treatment effects on the awareness of hypoglycemia ("blood sugar low") were noted. No change occurred in the score for this during euglycemia, but the mean +/- SE increase was 2.1 +/- 0.4 during hypoglycemia. This increase was not reduced significantly by adrenergic blockade (1.6 +/- 0.5), but was reduced significantly and substantially (approximately 70%) by pan-autonomic blockade (0.6 +/- 0.3). Significant neurogenic symptoms included shaky/tremulous (P < 0.001), heart pounding (P < 0.001), and nervous/anxious (P = 0.002), all adrenergic; and sweaty (P < 0.001), hungry (P < 0.001), and tingling (P = 0.009), all cholinergic. Significant neuroglycopenic symptoms, those produced by hypoglycemia but not reduced by pan-autonomic blockade, included warm (P < 0.001), weak (P = 0.011), difficulty thinking/confused (P = 0.004), and tired/drowsy (P = 0.003). We conclude that muscarinic cholinergic mechanisms mediate an important and previously uncharacterized component of the neurogenic symptoms of hypoglycemia and awareness of hypoglycemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243826 TI - Differential expression of GAD65 and GAD67 in human, rat, and mouse pancreatic islets. AB - The smaller form of the autoantigen glutamic acid decarboxylase, GAD65 (formerly the 64,000 M(r) autoantigen), is a major target of humoral autoimmunity in type I diabetes. Human autoantisera have been used extensively to characterize the GAD65 antigen in both rat and human islets, but the protein has escaped detection in mouse islets. We have now analyzed the expression of GAD65 and GAD67, the larger glutamic acid decarboxylase protein, in human, rat, and mouse islets of Langerhans and brain, using human monoclonal islet cell autoantibodies, human autoantisera, and experimentally raised antibodies to glutamic acid decarboxylase. Human monoclonal autoantibodies and experimentally raised antibodies reacted with mouse GAD65 produced in a baculovirus expression system by Western blotting and immunoprecipitation and with GAD65 in mouse brain by immunohistochemistry but failed to detect GAD65 in mouse islets by the latter two methods. However, analysis of mouse islets by Western blotting technique, using the most sensitive experimentally raised antibody, showed that mouse islets express both GAD65 and GAD67 but at levels that are severalfold lower than those in mouse brain or in human and rat islets. Furthermore, both human and rat islets predominantly express GAD65, whereas GAD67 is the major glutamic acid decarboxylase protein in mouse islets. Human islets are significantly distinct from mouse and rat islets and from brain because they only express GAD65, which is consistent with the predominant role of this form as a target of autoantibodies associated with beta-cell destruction in humans. Human as well as rat islet GAD65 are found in both membrane-bound and soluble forms. The low level of glutamic acid decarboxylase expression in mouse islets compared with human and rat islets is likely to have implications for both the development of tolerance to glutamic acid decarboxylase as well as the homing of glutamic acid decarboxylase-specific lymphocytes to the mouse beta-cell. In this context, the results suggest 1) that the mouse is ideal for studies of the consequences of an expression of high levels of glutamic acid decarboxylase in the beta-cell from a transgene and 2) that the rat may be better suited than the mouse for development of nontransgenic animal models of glutamic acid decarboxylase autoimmunity by immunization. PMID- 8243827 TI - Primary prevention of diabetes mellitus by prevention of obesity in monkeys. AB - Many, but not all, adult rhesus monkeys spontaneously develop significant increases in body fat mass, and many, but not all, progress to develop overt adult-onset type II diabetes. The purpose of this study was to determine whether both an increase in body fat and onset of diabetes could be simultaneously prevented through long-term maintenance of stable normal adult body weight by caloric titration. Eight adult male monkeys were provided a complete normal chow diet, but with daily amounts restricted and titrated on a weekly basis to maintain a constant body weight (weight-stabilized group). This regimen has been continued for 5-9 yr (mean +/- SD of 7 +/- 0.5 yr) with monkeys attaining the age of 17.9 +/- 0.6 yr and with maintenance of normal body fat (17.7 +/- 1.8%). The age-matched ad libitum fed group (18.1 +/- 0.2 yr of age) consisted of 19 monkeys maintained under identical laboratory conditions and diet, but with food available ad libitum. Results showed weight-stabilized monkeys weighed significantly less than ad libitum fed monkeys (10.4 +/- 0.2 vs. 16.1 +/- 0.7 kg, respectively, P < 0.05) and had significantly better glucose tolerance as measured by Kglucose (glucose disappearance rate) (3.9 +/- 0.3 vs. 2.4 +/- 0.2, P +/- 0.05). Of the 19 ad libitum fed age-matched monkeys, 4 were overtly diabetic, and 6 others had significantly reduced glucose tolerance. Hyperinsulinemia did not develop in the weight-stabilized group, and beta-cell response to glucose remained normal; both were significantly different from the exaggerated levels of the ad libitum fed group (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243828 TI - Lymph node T-cells do not optimally transfer diabetes in NOD mice. AB - The nonobese diabetic mouse in a model of spontaneous development of autoimmune type I diabetes. The disease can be induced in young, irradiated recipients by injecting splenic T-cells from diabetic donors. The adoptive transfer of diabetes requires the presence of both CD4+ and CD8+ splenic T-cell subsets. To test whether diabetogenic cells distribute in other lymphoid organs of diabetic mice, we first analyzed lymph node cells. Lymph node cells were much less efficient in transferring diabetes than splenocytes. This inefficacious transfer was not attributable to the absence of hematopoietic precursors or a lack of macrophages. Lymph node cells did not protect from the transfer of diabetes by splenocytes, indicating the absence of suppressor cells. Although CD8+ lymph node T-cells seemed functionally comparable to CD8+ splenocytes, CD4+ lymph node T-cells failed to cooperate with CD8+ splenocytes to transfer diabetes. Our study suggests that diabetogenic cells are not evenly distributed in the different lymphoid organs. This may reflect a differential migration pattern of pathogenic T-cells in this animal model. PMID- 8243829 TI - Expression of functional nerve growth factor receptors in pancreatic beta-cell lines and fetal rat islets in primary culture. AB - Previous data demonstrated that one rat insulinoma cell line, RINm5F cells, which is a rat beta-cell line derived from a pancreatic tumor, express mRNA coding for both the low- and the high-affinity nerve growth factor receptors. Goals of this study were to extend our data to other beta-cell lines and fetal islets in primary culture and to study further the binding characteristics of nerve growth factor receptors on beta-cells. Northern blot analysis revealed that not only a panel of endocrine beta-cell lines (RINm5F, INS-1, beta-TC3) but also fetal rat islets in primary culture express mRNA coding for trk-A, which has been proposed to be the neuronal high-affinity nerve growth factor receptors. Reverse polymerase chain reaction followed by sequencing revealed that the sequence of trk-A receptor in RINm5F cells is identical to that of trk-A expressed in PC12 cells. The expression of the low-affinity nerve growth factor receptor was examined by Northern blot analysis that showed low-affinity nerve growth factor receptor to be expressed in RINm5F and INS-1 cell lines, in fetal rat islets in primary culture, but not in beta-TC3-cells. Binding experiments revealed the presence of low- and high-affinity nerve growth factor binding sites, identical to those described for PC12 cells, on RINm5F and INS-1 cells and only high affinity binding sites on beta-TC3 cells. Exposure of all three beta-cell lines to nerve growth factor increased NGFI-A and c-fos mRNA steady-state levels, showing that these receptors are functional.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243830 TI - Ala1048-->Asp mutation in the kinase domain of insulin receptor causes defective kinase activity and insulin resistance. AB - We identified a heterozygous missense mutation that substituted aspartic acid (GAC) for alanine (GCC) at codon 1048 of the insulin receptor gene in a patient who displayed typical symptoms of Type A syndrome of insulin resistance. The proband's mother and younger brother were also found to be heterozygous for the mutation. We constructed the identified mutant insulin receptor cDNA by site directed mutagenesis, transfected the mutant cDNA into COS 7 cells, and found that kinase activity of the mutant insulin receptors was markedly impaired. Ala1048 is located in the kinase domain of the insulin receptor beta-subunit and is conserved in most of protein-tyrosine kinases. Besides, neighboring Glu1047 is invariant in all protein kinases and is thought to be involved in interaction with ATP. Photoaffinity labeling of the mutant insulin receptor with ATP analogue, 8-azido (alpha-32P)ATP was not influenced by the mutation, suggesting that the mutation did not inhibit ATP binding but possibly interfered with subsequent phosphoryl transfer. Insulin-stimulated phosphorylation of exogenous substrate by partially purified insulin receptors prepared from COS 7 cells that were cotransfected with wild-type and mutant insulin receptor cDNAs was markedly impaired, whereas autophosphorylation was decreased by approximately 50% of wild type receptors. These results indicated that the identified heterozygous substitution of Asp for Ala1048 in insulin receptor was responsible for insulin resistance of this patient. PMID- 8243831 TI - Treatment with an interleukin-1 receptor antagonist protein prolongs mouse islet allograft survival. AB - An interleukin-1 receptor antagonist protein was evaluated with regard to its efficacy in allogeneic and xenogeneic islet transplantation. Alloxan-induced diabetic C57BL/6 (H-2b) mice were transplanted under the kidney capsule with 500 C57BL/Ks (H-2d) mouse islets. Alzet osmotic pumps, which release their content over an 11- to 13-day period, were implanted subcutaneously for continuous infusion of interleukin-1 receptor antagonist protein (1.0, 5.0, 8.0 mg.kg-1 x day-1) or phosphate-buffered saline. Blood glucose determinations were performed every second or third day; at death, the islet-bearing kidneys were morphologically evaluated. Mice treated initially with the higher interleukin-1 receptor antagonist protein concentrations were followed for an additional period after cessation of the drug release to evaluate whether a transitory interleukin 1 receptor antagonist protein treatment would induce tolerance to the graft. All phosphate-buffered saline-treated mice were hyperglycemic 11 days after islet allotransplantation. Most of their grafts were heavily infiltrated with mononuclear cells. In the various interleukin-1 receptor antagonist protein treated groups, 60-80% of the mice were normoglycemic after 11 days. Moreover, light microscopic examinations showed that most mice treated with interleukin-1 receptor antagonist protein had normal islet grafts or grafts infiltrated with only a few mononuclear cells. After interruption of interleukin-1 receptor antagonist protein infusion (8.0 mg.kg-1 x day-1), all animals developed hyperglycemia within 2-9 days. In xenogeneic experiments, 500-750 fetal porcine islet-like cell clusters were transplanted under the kidney capsule of normoglycemic C57BL/6 mice. These animals were treated either with interleukin-1 receptor antagonist protein (8.0 mg.kg-1 x day-1) or phosphate-buffered saline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243832 TI - The sulfonylurea drug, glimepiride, stimulates glucose transport, glucose transporter translocation, and dephosphorylation in insulin-resistant rat adipocytes in vitro. AB - Sulfonylurea drugs are widely used in the therapy of NIDDM. The improvement of glucose tolerance after long-term treatment of NIDDM patients with the drug can be explained by stimulation of glucose utilization in peripheral tissues that are characterized by insulin resistance in these patients. We studied whether the novel sulfonylurea drug, glimepiride, stimulates glucose transport into isolated insulin-resistant rat adipocytes. After long-term incubation of the cells in primary culture with high concentrations of glucose, glutamine, and insulin, stimulation of glucose transport by insulin was significantly reduced both with respect to maximal responsiveness (65% decrease of Vmax) and sensitivity (2.6 fold increase of ED50) compared with adipocytes cultured in medium containing a low concentration of glucose and no insulin. This reflects insulin resistance of glucose transport. In contrast, both responsiveness and sensitivity of glucose transport toward stimulation by glimepiride were only marginally reduced in insulin-resistant adipocytes (15% decrease of Vmax; 1.2-fold increase of ED50) versus control cells. Glimepiride, in combination with glucose and glutamine during the primary culture, caused desensitization of the glucose transport system toward stimulation by insulin, but to a lesser degree than insulin itself (50% reduction of Vmax; ninefold increase of ED50). Again, the maximal responsiveness and sensitivity of glucose transport toward stimulation by glimepiride were only slightly diminished. The presence of glimepiride during primary culture did not antagonize the induction of insulin resistance of glucose transport. The stimulation of glucose transport in insulin-resistant adipocytes by glimepiride is caused by translocation of glucose transporters from low density microsomes to plasma membranes as demonstrated by subcellular fractionation and immunoblotting with anti-GLUT1 and anti-GLUT4 antibodies. Immunoprecipitation of GLUT4 from 32Pi- and [35S]methionine-labeled adipocytes revealed that the insulin resistance of GLUT4 translocation is accompanied by increased (three- to fourfold) phosphorylation of GLUT4 in both low-density microsomes and plasma membranes. Short-term treatment of desensitized adipocytes with glimepiride or insulin reduced GLUT4 phosphorylation by approximately 70 and 25%, respectively, in both fractions. We conclude that glimepiride activates glucose transport by stimulation of GLUT1 and GLUT4 translocation in rat adipocytes via interference at a site downstream of the putative molecular defect in the signaling cascade between the insulin receptor and the glucose transport system induced by high concentrations of glucose and insulin. The molecular site of glimepiride action is related to GLUT4 phosphorylation/dephosphorylation, which may regulate glucose transporter activity and translocation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243833 TI - Different sensitivity of glucose and amino acid metabolism to insulin in NIDDM. AB - NIDDM subjects are characterized by impaired glucose tolerance and insulin resistance with respect to glucose metabolism. To examine whether the defect in glucose utilization extends to amino acid metabolism, 6 NIDDM subjects (64 +/- 4 yr of age; ideal body weight of 107 +/- 3%) and 7 control subjects (58 +/- 4 yr of age; ideal body weight of 105 +/- 2%) were studied with the euglycemic insulin clamp technique, in combination with [1-14C]leucine and indirect calorimetry. All subjects participated in two studies. In study 1, after 3 h of tracer equilibration, a 3-h insulin clamp (40 mU.m-2 x min-1) was performed to define the effect of insulin on leucine kinetics and glucose metabolism. In study 2, subjects received a repeat 3-h insulin clamp, and a balanced amino acid solution was infused to increase the plasma amino acid concentrations approximately 2-fold to examine the effect of combined physiological hyperinsulinemia hyperaminoacidemia on the rate of leucine and glucose disposal. Insulin-mediated total body glucose uptake was significantly reduced in NIDDM during both study 1 (5.6 +/- 0.4 vs. 6.9 +/- 0.6 mg.kg-1 x min-1, P < 0.01) and study 2 (5.2 +/- 0.4 vs. 6.8 +/- 0.6, P < 0.01). Basal plasma leucine (120 +/- 10 vs. 123 +/- 11 microM) and alpha-ketoisocaproic acid concentrations (28 +/- 3 vs. 25 +/- 2 microM) were similar in NIDDM and control subjects, respectively. In contrast, the basal plasma glucose concentration (8.9 +/- 0.8 vs. 4.7 +/- 0.2 microM) and the HbA1c (8.5 +/- 0.2 vs. 5.7 +/- 0.2%) were significantly increased in NIDDM (P < 0.01). In the postabsorptive state, endogenous leucine flux, leucine oxidation, and nonoxidative leucine disposal were similar in NIDDM and control subjects. When insulin was infused without amino acids (study 1), the decrement in plasma leucine (53 +/- 5 vs. 48 +/- 4 microM), endogenous leucine flux (13 +/- 2 vs. 11 +/- 1 mumol.m-2 x min-1), leucine oxidation (1.6 +/- 0.2 vs. 1.3 +/- 0.1 mumol.m 2 x min-1), and nonoxidative leucine disposal (10 +/- 1 vs. 8 +/- 1 mumol.m-2 x min-1) was comparable in both groups. During combined insulin and amino acid infusion (study 2), plasma leucine concentration (185 +/- 20 vs. 190 +/- 15 microM) rose similarly in NIDDM and control subjects.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8243834 TI - G-protein specificity in signaling pathways that mobilize calcium in insulin secreting beta-TC3 cells. AB - Fuel- and receptor-induced signal transduction pathways were investigated in beta TC3 cells, an insulin-secreting cell line. An increase of glucose concentration from 0 to 15 mM and stimulation with 0.5 mM carbachol resulted in up to a twofold increase in insulin secretion by beta-TC3 cells, and their simultaneous addition increased insulin release eightfold. In single fura 2-loaded cells, a potentiating effect of carbachol was also observed on glucose-induced intracellular Ca2+ mobilization. Immunoblotting with antibodies raised to the COOH-terminal of G-protein alpha-subunits showed that G alpha i, G alpha o, and G alpha q are present in beta-TC3 cells in commensurable quantities. The novel technique of microinjection of anti-G alpha antibodies into single beta-cell was used to probe the functional role of these G-proteins. Microinjection of anti-G alpha i antibodies into beta-TC3 cells had no effect on glucose- and carbachol induced Ca2+ mobilization. However, anti-G alpha q completely inhibited the Ca(2+)-mobilizing effect of carbachol, but not of glucose, within 5 min. Microinjection of anti-G alpha o antibodies had no effect on carbachol-induced Ca2+ mobilization. Microinjection of anti-G alpha i and anti-G alpha q antibodies had a minimal effect on glucose-induced Ca2+ mobilization (< 8% of cells nonresponding), but microinjection of anti-G alpha o increased the proportion of nonresponding cells to 37%. The results suggest that, in beta-TC3 cells, distinct signal transduction pathways with specific G-protein involvement may interact with secretagogue-induced Ca2+ mobilization and, ultimately, with insulin secretion. PMID- 8243835 TI - Importance of various intracellular regulatory mechanisms of polyamine metabolism in camostate-induced pancreatic growth in rats. AB - Aim of the present study was to evaluate the relevance of various intracellular regulatory mechanisms of polyamine metabolism in normal and camostate (FOY 305) induced pancreatic growth in rats. Furthermore it was investigated whether the simultaneous inhibition of the polyamine interconversion pathway by the potent polyamine oxidase inhibitor MDL-72527 together with the ornithine decarboxylase (ODC) inhibitor alpha-difluoromethylornithine (DFMO) is able to enhance and prolong the only initial and transient inhibitory effects of DFMO on camostate induced pancreatic growth. Simultaneous administration of DFMO and MDL-72527 resulted in a significant inhibition of the camostate-induced increases in pancreatic putrescine, ODC and DNA over 5 days, while the initial significant inhibition of pancreatic weight, protein content, DNA-polymerase and especially spermidine was absent after 5 days and spermine as well as N1-acetylspermidine were even increased. Diamine oxidase (DAO) activity in pancreas is very low and the potent DAO inhibitor aminoguanidine failed to alter any of the measured parameters except DAO. Therefore, oxidative degradation of putrescine by DAO- which is essential in the gut--is irrelevant in the pancreas. Simultaneous inhibition of the pancreatic polyamine interconversion pathway extended the initial inhibitory effects of DFMO only slightly and failed to exert a potent long-lasting inhibitory effect on spermidine and pancreatic growth. PMID- 8243836 TI - Ulcerative colitis serum recognizes the M(r) 40K protein on colonic adenocarcinoma cells for antibody-dependent cellular cytotoxicity. AB - Antibodies in ulcerative colitis (UC) serum promote antibody-dependent cellular cytotoxicity (ADCC) against colon cancer cells. The target molecules on the cells recognized by the antibody are unknown. This study examines the role of the M(r) 40K colon epithelial protein in this process. Serum from 29 UC patients, but not from 19 patients with Crohn's disease, significantly (p < 0.001) induced more cytotoxicity than did normal serum against DLD-1 colon cancer cells which are known to express the M(r) 40K protein. Peripheral blood lymphocytes but not polymorphonuclear leukocytes showed ADCC. Anti-M(r) 40K monoclonal antibodies significantly (p < 0.001) inhibited the UC-serum-induced ADCC, but not the spontaneous cytotoxicity. These results suggest the involvement of the M(r) 40K protein in ADCC induced by UC serum. PMID- 8243837 TI - Molecular forms of pseudocholinesterase in patients with Crohn's disease. AB - Previous studies have shown that pseudocholinesterase (PCHE) is low in patients with Crohn's disease (CD). This finding, however, failed to be of clinical relevance due to the wide normal range. PCHE consists of four main molecular forms designated as C1, C2, C3 and C4 according to their electrophoretic mobility. The question of the present study was to assess the influence of CD on the distribution and pattern of the PCHE isozymes. We therefore investigated the electrophoretic separation of PCHE in 16 healthy volunteers (HV), in 15 patients with quiescent CD (QCD; CD activity index: median = 71, interquartile range = 44 122) and in 10 patients with active CD (ACD; CD activity index: 229, 173-304). In most of the cases total serum PCHE activity was within the normal range even in patients with active CD. No changes of the isozyme pattern were found but the percentage distribution was significantly influenced by the inflammatory activity in patients with active CD:C1 (HV: 14.7%, 13.7-18.1%; QCD: 16.0%, 9.8-19.9%; ACD: 8.5%, 2.9-12.5%, p < 0.01) and C2 (HV: 8.0%, 6.7-10.5%; QCD: 9.0%, 7.9-9.7%; ACD: 6.7%, 3.2-8.6%, p < 0.05) were decreased in active CD while the C4 component (HV: 66.8%, 62.5-69.9%; QCD: 63.1%, 54.9-73.8%; ACD: 77.3%, 70.7-90.1%, p < 0.01) was increased. The percentage of the C3 band (HV: 5.7%, 4.6-6.9%; QCD: 6.8%, 4.6 8.4%; ACD: 5.3%, 2.8-6.8%, NS) was unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243838 TI - Impairment of bacterial flora in human ulcerative colitis and experimental colitis in the rat. AB - Changes in the colonic mucosa-associated microflora were determined both in patients with active and inactive ulcerative colitis and in rats with acetic acid induced colitis. In patients with active ulcerative colitis, significant decreases in the number of anaerobic bacteria (Brain Heart Infusion medium), anaerobic gram-negatives and Lactobacillus were found, whereas no changes were seen in the number of aerobic bacteria and Enterobacteriaceae. In patients with inactive ulcerative colitis, no significant differences in colonic mucosa associated microflora could be demonstrated. Similar changes were seen in rats with acetic acid-induced colitis. Thus, 4 days after acetic acid administration, at which time the colitis was well developed as evaluated by morphological appearance and myeloperoxidase activity, reduction in the number of anaerobic bacteria and Lactobacillus was seen. The first day after acetic acid administration, when the colitis had not developed, or after 14 days, when the colitis had been overcome, no alterations were seen in the mucosa-associated microflora as compared with control rats. We conclude that a reduction in the number of anaerobic bacteria and Lactobacillus is a common feature in active colitis regardless of the origin of colitis. PMID- 8243839 TI - Perceived sexual dysfunction amongst patients with inflammatory bowel disease. AB - 300 patients with inflammatory bowel disease (IBD) were randomly selected from the community-based register held in Leicester. They were invited to participate in a study investigating personal problems of patients with IBD. 188 patients agreed to participate and were subsequently sent a previously validated questionnaire. A similar questionnaire was sent to buddy controls of similar age, sex and background (n = 76) as well as matched controls drawn at random from general practitioner lists (n = 46). Details sought included demographic data, patients' perception of their disease severity, data relating to treatment, family history, fertility, frequency of sexual intercourse and the effect IBD had on personal relationships. 150 questionnaires were returned from patients (response rate 80%) and 122 from controls. The reliability of a randomly selected group of questionnaires was assessed by direct interview 4 months after the main study (n = 20). There were no demographic differences between the patient and control groups (age: t = 0.67, ns; duration of marriage: t = 0.92, and marital status/number with a regular partner: chi 2 = 14, 12 d.f., p = 0.3). Dyspareunia was commoner amongst patients (n = 15, 38%) than controls (n = 7, 18%), although this was not statistically significant (z = 2.6). There was no significant difference in the overall frequency of sexual intercourse amongst patient and control groups (chi 2 = 12.78, 12 d.f.), even when comparing those with infrequent or no sexual intercourse (chi 2 = 6.98, 4 d.f.) The reliability of these results was confirmed after 4 months when there was no difference in responses in the re-interviewed group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243840 TI - The 1993 Claude Bernard lecture. Domenico Andreani, Rome, Italy. PMID- 8243841 TI - The 28th Minkowski prize winner 1993. Hannele Yki-Jarvinen, Helsinki, Finland. PMID- 8243842 TI - 1993 Castelli Pedroli prize, 8th Camillo golgi lecture. Kristian Folkvord Hanssen, Oslo, Norway. PMID- 8243843 TI - The Albert Renold fellowship 1993 (I). Vincent Poitout, Paris, France. PMID- 8243844 TI - The Albert Renold fellowship 1993 (II). Carani B. Sanjeevi, Stockholm, Sweden. PMID- 8243845 TI - 1993 Mollegaard BB rat travel fellowship. Anna Pettersson, Charlestown, Massachusetts, USA. PMID- 8243846 TI - The Eli Lilly/EASD research fellowship 1993. Decio L. Eizirik, Uppsala, Sweden. PMID- 8243847 TI - Is microalbuminuria part of the prediabetic state? The Mexico City Diabetes Study. AB - Microalbuminuria is associated with increased cardiovascular mortality in both diabetic and non-diabetic subjects. A number of studies have indicated that insulin resistance, increased blood pressure and dyslipidaemia precede the onset of clinical diabetes. We examined various correlates of microalbuminuria in 1,298 non-diabetic subjects who participated in the Mexico City Diabetes Study, a population-based study of diabetes and cardiovascular risk factors. Both parental history of diabetes and impaired glucose tolerance were significantly associated with microalbuminuria. These results were not explained by differences in age or blood pressure between subjects with or without a parental history of diabetes or impaired glucose tolerance. In addition, subjects with microalbuminuria had increased 2-h insulin and triglyceride concentrations, a higher prevalence of hypertension, and decreased high density lipoprotein cholesterol concentrations relative to subjects without microalbuminuria. These results that microalbuminuria may be a feature of the prediabetic state. PMID- 8243848 TI - Epidemiology of renal involvement in newly-diagnosed middle-aged and elderly diabetic patients. Cross-sectional data from the population-based study "Diabetes Care in General Practice", Denmark. AB - We report on a study in which 487 Danish general practitioners participated with the purpose of including all newly-diagnosed diabetic patients aged 40 years or more from a well-defined catchment population during a well-defined time period. A total of 1267 diabetic patients with a median age of 65.3 years were included. Renal involvement was assessed from the albumin/creatinine ratio in a morning urine sample. Albumin/creatinine ratio was < 2/2- < 20/ > or = 20 mg/mmol in 59.8/33.6/6.6% of male and 66.6/28.8/4.6% of female patients. The level of albumin/creatinine ratio increased with age and the observed overall male predominance was almost confined to diabetic patients with an albumin/creatinine ratio of 5 mg/mmol or greater. By taking into account the confounding effect of age and sex, a positive association between smoking and albumin/creatinine ratio was disclosed. Moreover, high systolic blood pressure, hypertriglyceridaemia, hypercholesterolaemia (males only) and high HbA1c, but not body mass index or diastolic blood pressure were identified as correlates of elevated albumin/creatinine ratio. Glucosuria was positively correlated with albumin/creatinine ratio even when the influence of HbA1c, sex and age was taken into account. A positive correlation between serum creatinine and albumin/creatinine ratio was seen in males, but not in females. In addition, renal involvement was associated with the presence of peripheral angiopathy and diabetic retinopathy and with high resting heart rate. The cross-sectional data presented highlight the importance of reducing the overall burden of modifiable risk factors in newly-diagnosed Type 2 diabetic patients. PMID- 8243849 TI - Microalbuminuria in a random cohort of recently diagnosed type 2 (non-insulin dependent) diabetic patients living in the greater Munich area. AB - Still under debate is the prevalence of microalbuminuria in patients with recently diagnosed Type 2 (non-insulin-dependent) diabetes mellitus and its relation to existing macro-vascular disease and the major vascular risk markers. Hence, from a representative sample of 1512 patients with Type 2 diabetes of varied duration (recruited from 22 non-specialized medical practices of the Greater Munich Area) 68 (26 males, 42 females) of 71 eligible subjects with a known duration of diabetes of up to 17 weeks and not less than 4 weeks were examined in the present study. Median age was 61 (39 to 75) years, prevalence of ischaemic heart disease (case history plus ECG, Minnesota code, Whitehall criteria) 41.2%, and that of peripheral vascular and carotid artery disease (both assessed by ultrasound-Doppler) were 35.3 and 4.4%, respectively. Diabetes was well controlled (HbA1c: 6.9%, 5.6-8.3; fasting blood glucose: 7.7 mmol/l, 5.4 10.4; median +/- interquartile range IQ), the cardiovascular risk profile was most prominent in terms of triglycerides (3.1 mmol/l, 2.1-4.6, median +/- IQ range) and systolic blood pressure (164 mm Hg, 140-186, median +/- IQ range). 13.2% showed signs of urinary tract infection. Of the remainder, 19.0% exhibited microalbuminuria (RIA, > 30-200 mg/l), and 5.2% macroalbuminuria (> 200 mg/l). Significant correlations (p < 0.05) were found between urinary albumin concentration and beta 2-microglobulin in serum, systolic blood pressure, serum triglycerides, serum HDL-cholesterol (inversely), HbA1c, and peripheral vascular disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243850 TI - UK Prospective Diabetes Study (UKPDS). X. Urinary albumin excretion over 3 years in diet-treated type 2, (non-insulin-dependent) diabetic patients, and association with hypertension, hyperglycaemia and hypertriglyceridaemia. AB - Urinary albumin excretion has been assessed in 585 newly-presenting Type 2 (non insulin-dependent) diabetic patients (aged 53 (8) years, 67% male) at diagnosis with fasting plasma glucose 10.3 (3.2) mmol/l and over 3 years of dietary treatment. Urinary albumin at diagnosis, geometric mean (1 SD interval) corrected for dilution by regression on urine creatinine concentration of 10 mmol/l, was 17 (5-58) mg/l compared with 8 (3-18) mg/l in an age-matched non-diabetic reference population. Values greater than 50 mg/l were found in 17% of diabetic patients compared with 4% in the reference group. After diet therapy for 3 months, fasting plasma glucose decreased to 6.9 mmol/l and urinary albumin to 12 (4-31) mg/l (p < 0.0001). This suggests that increased urinary albumin excretion at diagnosis is in part functional, possibly secondary to glomerular hyperfiltration caused by hyperglycaemia and raised blood pressure. Over the next 3 years, mean fasting plasma glucose was 7.2 mmol/l, albumin excretion changed little, without significant increase either in patients with raised or normal albumin at diagnosis. Both at diagnosis and over 3 years, urinary albumin excretion was independently associated with fasting plasma glucose and triglyceride levels and with systolic blood pressure, but the combination of these factors only explained 10% of the total variance. This suggests the presence of additional pathological processes in patients with increased urinary albumin. Urinary albumin was not associated with other variables included in syndrome X, such as HDL cholesterol, fasting plasma insulin, obesity or central adiposity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243851 TI - Prognostic value of urinary albumin excretion rate and other risk factors in elderly diabetic patients and non-diabetic control subjects surviving the first 5 years after assessment. AB - In 1981-1982 urinary albumin excretion rates were determined in 211 diabetic and 216 non-diabetic subjects aged 60-74 years. By April 1992 122 diabetic and 58 non diabetic probands had died. Dividing the two study populations at an albumin excretion rate of 15 micrograms/min showed that 69.3% of diabetic subjects with values at or above the limit, and 49.9% of those with values below (log rank test p = 0.0082) had died. The corresponding values for non-diabetic subjects were 44.4% and 21.0%, respectively (log rank test p = 0.0002). In single factor log rank tests ischaemic heart disease and a low value of HDL were also predictive of death in the diabetic population during a 10-11-year observation period. In the non-diabetic population ischaemic heart disease, hypertension, and a serum creatinine level above the median value were predictive. In further log rank analyses probands dying during the first years, (e.g. the first 2 years) were removed from the calculations. The prognostic value of the above-mentioned factors diminished with time. In a Cox Regression analysis we found that the predictive value of urinary albumin excretion rate to mortality had disappeared when subjects who had died during the first 5 years were removed from the analysis, whereas HDL in the diabetic patients and blood pressure and serum creatinine in non-diabetic subjects were still of significant predictive value. We therefore conclude that urinary albumin excretion rate is a more short-term predictor of mortality than previously thought, in contrast to HDL, hypertension and serum creatinine. PMID- 8243852 TI - Relationship of progressively increasing albuminuria to apoprotein(a) and blood pressure in type 2 (non-insulin-dependent) and type 1 (insulin-dependent) diabetic patients. AB - This study has explored the temporal relationship between apoprotein(a), blood pressure and albuminuria over a mean interval of 11 years in a cohort of 107 diabetic patients of whom 26 (14 Type 2 (non-insulin-dependent), 12 Type 1 (insulin-dependent) had progressively increasing albuminuria ('progressors'). In Type 2 diabetic patients, no significant differences were noted for HbA1, blood pressure, creatinine clearance or serum lipids between progressors and non progressors. In Type 1 diabetic patients, final systolic and diastolic blood pressures were higher in progressors compared with non-progressors and progressors showed impairment of renal function in association with a rise in blood pressure at the macroalbuminuric stage. Initial apoprotein(a) levels were similar in progressors and non-progressors of either diabetes type. Apoprotein(a) levels increased exponentially with time in 12 of 14 Type 2 progressors but only in 5 of 12 Type 1 progressors (p < 0.01). In Type 2 diabetic patients, the annual increase in apoprotein(a) levels was 9.1 +/- 2.4%, which was significantly greater than in non-progressors, 2.0 +/- 1.2% (p < 0.01) and also exceeded the rates of increase of apoprotein(a) in progressors with Type 1 diabetes, 4.0 +/- 1.4%, (p < 0.05). Apoprotein(a) levels correlated significantly with albuminuria in 8 of 14 Type 2 progressors but only in 3 of 12 Type 1 progressors (p < 0.05). The rate of increase of apoprotein(a) levels was not related to mean HbA1, creatinine or creatinine clearance levels, or to albuminuria.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243853 TI - Thrombogenic factors are related to urinary albumin excretion rate in type 1 (insulin-dependent) and type 2 (non-insulin-dependent) diabetic patients. AB - Parameters of haemostasis, endothelial cell markers and lipid peroxide levels were studied in 64 Type 1 (insulin-dependent) and 94 Type 2 (non-insulin dependent) diabetic patients according to their urinary albumin excretion rate in comparison with age-matched control subjects. We determined plasma levels of fibrinogen (Clauss' method), coagulation factor VII:activity (clotting assay), factor VII antigen, protein C and S antigen, von Willebrand factor antigen, D dimer concentration (ELISA), and lipid peroxide levels (thiobarbituric acid) in relation to urinary albumin excretion rate (RIA). Significant positive correlations were found between urinary albumin excretion rate and plasma fibrinogen (p < 0.005, p < 0.02), factor VII activity (p < 0.0002, p < 0.002), factor VII antigen (p < 0.0001, p < 0.001), protein C (p < 0.003, p < 0.05), and lipid peroxides (p < 0.02, p < 0.004) in Type 1 as well as in Type 2 diabetes. Von Willebrand factor (p < 0.001) and protein S (p < 0.0005) correlated with albuminuria only in patients with Type 1 diabetes. Although most of the haemostatic abnormalities are already found in normoalbuminuric patients, the significant positive correlations to urinary albumin excretion indicate that endothelial cell damage and coagulation disorders deteriorate with the progression of diabetic nephropathy. PMID- 8243854 TI - Effect of metabolic factors and blood pressure on kidney function in proteinuric type 2 (non-insulin-dependent) diabetic patients. AB - Decline of kidney function with time and its influencing factors were investigated in the present longitudinal study in Type 2 (non-insulin-dependent) diabetic patients with clinical diabetic nephropathy. Compared to a control group of Type 2 diabetic patients without proteinuria, the proteinuric patients showed a higher prevalence of hypertension, higher systolic blood pressure values and serum triglyceride levels. The annual loss of glomerular kidney function was much higher in the proteinuric patients (5.3 ml.min-1 x 1.73 m2) than in the control subjects (0.9 ml.min-1 x 1.73 m2). Correlation analyses revealed a close correlation between the annual decrease of kidney function and the factors, systolic and diastolic blood pressure, triglyceride and postprandial blood glucose level as well as body mass index. Regression analyses showed for the first time that in addition to the systolic blood pressure and metabolic control, the triglyceride level is also an independent factor influencing the progression of nephropathy. Higher values of these parameters were associated with a more rapid deterioration of kidney function. PMID- 8243855 TI - Glomerulosclerosis in type 2 (non-insulin-dependent) diabetes mellitus: relationship to glycaemia in the University Group Diabetes Program (UGDP). AB - Kidney tissue of acceptable quality was available from autopsies of 55 patients who had been followed prospectively for 3 to 15 years as participants in the University Group Diabetes Program, a study of vascular disease in Type 2 (non insulin-dependent) diabetic patients. Slides were prepared for light microscopic reading by uniform histologic techniques, and then were randomly intermixed and coded with tissues identically prepared from matched non-diabetic subjects (morphologic controls). After independent review by three morphologists, the results were tabulated and assigned to one of four diagnostic groups: 1) typical diabetic nodular glomerulosclerosis; 2) mesangial changes suggestive of diabetes (diffuse lesion); 3) non-diabetic renal disease; 4) normal for age. Of the diabetic cases 31% (17 of 55) were found to show nodular glomerulosclerosis, and another 47% (26 of 55) showed suggestive changes; none of the morphologic control slides was read as showing nodular glomerulosclerosis, but some were judged to show suggestive mesangial (diffuse) changes. Although only 4 of the 17 diabetic patients with nodules had died of uraemia, many had hypertension, which may have contributed to their deaths from vascular disease. The patients with nodular glomerular changes also showed, on the average, the highest blood glucose levels during life. Type 2 diabetes in later life appears to be associated with a high risk for typical tissue changes of diabetic kidney damage, which may contribute significantly to morbidity and mortality and may be present before azotaemia and qualitative proteinuria have been recognized. PMID- 8243856 TI - Glomerular structure and function in proteinuric type 2 (non-insulin-dependent) diabetic patients. AB - Glomerular ultrastructure was examined in a series of 20 Type 2 (non-insulin dependent) diabetic patients with proteinuria. Reference was made to data previously obtained in non-diabetic kidney donors and in Type 1 (insulin dependent) diabetic patients with similar degrees of proteinuria. The Type 2 diabetic patients demonstrated the changes which characterize the diabetic glomerulopathy seen in Type 1 diabetic patients: basement membrane thickening, and increase in the mesangium and mesangial matrix expressed as fraction of the glomerular volume. Among the Type 2 diabetic patients there was more variation than among the Type 1 diabetic patients, as this group included subjects with normal parameters. The group means and coefficients of variation (= SD/mean) of the glomerulopathy parameters combined in the glomerulopathy index = basement membrane thickness/10+ Vv(matrix/glom).100 were 81 (0.30) and 92 (0.15) in the two diabetic groups, clearly different from the non-diabetic index, 42 (0.16). All Type 2 diabetic patients who also had retinopathy had a glomerulopathy index above the normal range. Similar changes in glomerular composition were seen in the two diabetic groups: with increasing glomerulopathy the volume of matrix dominated over the peripheral basement membrane, and a shift in the ratio of interfaces was seen: mesangial surface towards capillary lumen increased relative to the urinary surface, and peripheral capillary surface comprised less of the total capillary surface. Data indicated marked glomerular hypertrophy, which correlated with the mesangial volume fraction, thus encompassing preserved filtration surface per glomerulus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243857 TI - The course of kidney function in type 2 (non-insulin-dependent) diabetic patients with diabetic nephropathy. AB - We evaluated the impact of some putative progression promoters on kidney function in albuminuric Type 2 (non-insulin-dependent) diabetic patients with biopsy proven diabetic glomerulosclerosis. Twenty-six patients (1 female) with a mean age of 52 (standard error 2) years and a known mean duration of diabetes of 9 (1) years were followed-up prospectively for a mean of 5.2 (range 1.0-7.0) years. Twenty-one patients received antihypertensive treatment. During the observation period the glomerular filtration rate decreased from 83 (24-146) to 58 (2-145) ml.min-1 x 1.73 m-2 (mean (range)) (p < 0.001). The mean rate of decline in glomerular filtration rate was 5.7 (-3.5 to 22.0) ml/min per year. Albuminuria increased from 1.2 (0.3-7.2) to 2.3 (0.4-8.0) g/24 h (geometric mean (range)) (p < 0.001). Arterial blood pressure remained unchanged: 162/93 (SE 4/3) and 161/89 (4/2) mm Hg. Univariate analysis showed the rate of decline in glomerular filtration rate to correlate with systolic blood pressure (r = 0.71, p < 0.001), mean blood pressure (r = 0.56, p < 0.005), albuminuria (r = 0.58, p < 0.005) and the initial glomerular filtration rate (r = -0.49, p < 0.02). The rate of decline in glomerular filtration rate did not correlate significantly with dietary protein intake, total cholesterol, high-density lipoprotein cholesterol or HbA1c. Three patients died from uraemia and four patients died from cardiovascular disease. Two patients required renal replacement therapy at the end of the observation period. Our prospective observational study revealed that one-fifth of the patients developed end-stage renal failure during the 5-year observation period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243858 TI - Renal function and insulin sensitivity during simvastatin treatment in type 2 (non-insulin-dependent) diabetic patients with microalbuminuria. AB - The effect of simvastatin (10-20 mg/day) on kidney function, urinary albumin excretion rate and insulin sensitivity was evaluated in 18 Type 2 (non-insulin dependent) diabetic patients with microalbuminuria and moderate hypercholesterolaemia (total cholesterol > or = 5.5 mmol.l-1). In a double-blind, randomized and placebo-controlled design treatment with simvastatin (n = 8) for 36 weeks significantly reduced total cholesterol (6.7 +/- 0.3 vs 5.1 mmol.l-1 (p < 0.01)), LDL-cholesterol (4.4 +/- 0.3 vs 2.9 +/- 0.2 mmol.l-1 (p < 0.01)) and apolipoprotein B (1.05 +/- 0.04 vs 0.77 +/- 0.02 mmol.l-1 (p < 0.01)) levels as compared to placebo (n = 10). Both glomerular filtration rate (mean +/- SEM) (simvastatin: 96.6 +/- 8.0 vs 96.0 +/- 5.7 ml.min-1 x 1.73 m-2, placebo: 97.1 +/- 6.7 vs 88.8 +/- 6.0 ml.min-1 x 1.73 m-2)(NS) and urinary albumin excretion rate (geometric mean x/divided by antilog SEM) (simvastatin: 18.4 x/divided by 1.3 vs 16.2 x/divided by 1.2 microgram.min-1, placebo 33.1 x/divided by 1.3 vs 42.7 x/divided by 1.3 micrograms.min-1)(NS) were unchanged during the study. A euglycaemic hyperinsulinaemic clamp was performed at baseline and after 18 weeks in seven simvastatin- and nine placebo-treated patients. Isotopically determined basal and insulin-stimulated glucose disposal was similarly reduced before and during therapy in both the simvastatin (2.0 +/- 0.1 vs 1.9 +/- 0.1 (NS) and 3.1 +/- 0.6 vs 3.1 +/- 0.7 mg.kg-1 x min-1 (NS)) and the placebo group (1.9 +/- 0.1 vs 1.8 +/- 0.1 (NS) and 4.1 +/- 0.6 vs 3.8 +/- 0.2 mg.kg-1 x min-1 (NS)).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243859 TI - Determinants of end-stage renal disease in Pima Indians with type 2 (non-insulin dependent) diabetes mellitus and proteinuria. AB - To identify factors related to the development of end-stage renal disease after the onset of proteinuria, its incidence was determined in 364 Pima Indians aged 35 years or older with Type 2 (non-insulin-dependent) diabetes mellitus and proteinuria (protein-to-creatinine ratio > or = 0.5 g/g). Of these 364 subjects, 95 (36 men, 59 women) developed end-stage renal disease. The cumulative incidence was 40% 10 years after and 61% 15 years after the onset of proteinuria. The incidence of end-stage renal disease was significantly related to the duration of diabetes, the duration of proteinuria, higher 2-h plasma glucose concentration, type of diabetes treatment, and the presence of retinopathy at the time of recognition of the proteinuria, but not to age, sex, or blood pressure. Duration of proteinuria influenced the risk of end-stage renal disease, contingent, however, upon the duration of diabetes at the onset of proteinuria. The higher cumulative incidence of end-stage renal disease 15 years after the onset of proteinuria in Pima Indians (61%) than in Caucasians from Rochester, Minnesota (17%) may be attributable, in part, to the younger age of onset of Type 2 diabetes in Pima Indians than in Caucasians, to ethnic differences in susceptibility to renal disease, or to lower death rates among the Pima Indians from competing causes of death, such as coronary heart disease. PMID- 8243860 TI - Comparison of the course to end-stage renal disease of type 1 (insulin-dependent) and type 2 (non-insulin-dependent) diabetic nephropathy. AB - Is the course leading to diabetic end-stage renal disease similar for Type 1 (insulin-dependent) and Type 2 (non-insulin-dependent) diabetes mellitus? We identified all diabetic end-stage renal disease patients starting renal replacement therapy from 1989 to 1991 in two urban counties in Texas. Three ethnic/racial groups were enrolled: Mexican Americans, non-Hispanic Whites, African Americans. Patients were interviewed and their medical records, both inpatient and out-patient, were abstracted for relevant diagnostic and therapeutic information. We attempted to obtain records as far back as the onset of diabetes or hypertension and from all physicians who had cared for the patient. An historical algorithm was used to determine diabetic type. Of the patients enrolled, 91 were Type 1 and 438 were Type 2 diabetic patients. Type 1 diabetic patients had higher mean glucose levels in the first 10 years of diabetes (16.3 vs 11.4 mmol/l) but lower systolic blood pressures (148 vs 157 mmHg). The duration of diabetes prior to end-stage renal disease was longer for Type 1 than Type 2 patients (22 vs 17 years). Type 1 diabetic patients were more likely to have other microvascular complications (retinopathy, neuropathy, gastroparesis), less likely to have coronary disease (myocardial infarction and congestive heart failure), and had similar rates of stroke and vascular surgery procedures (carotid endarterectomy, coronary artery bypass surgery, aortofemoral bypass). Type 1 and Type 2 diabetic patients were just as likely to have a first degree relative with hypertension (60.5 vs 65.5%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243861 TI - Epidemiology, development and treatment of end-stage renal failure in type 2 (non insulin-dependent) diabetic patients in Europe. AB - The aim of the present report was to compare the current patterns of incidence and prevalence of end-stage renal failure and mode of renal replacement therapy in patients with Type 1 (insulin-dependent) and Type 2 (non-insulin-dependent) diabetes mellitus in Europe. All Type 1 and Type 2 diabetic patients recorded on the Registry of the European Dialysis and Transplant Association as being alive on renal replacement therapy were analysed according to age, sex, geographic distribution, and mode of therapy (haemodialysis, peritoneal dialysis or renal transplantation). During 1990 3981 diabetic patients commenced renal replacement therapy in Europe, and at 31 December 1990 a total of 15,197 diabetic patients were receiving treatment. One-third were reported to be Type 2 diabetic patients, but the true proportion is expected to be higher. Both male and female Type 2 diabetic patients were older than Type 1 patients. Major geographic variations were observed; annual acceptance of Type 2 diabetic patients for treatment was greatest in Austria (10.7 per million) and equal to Type 1 patients, whereas the number of Type 1 diabetic patients was four times that of Type 2 patients in Sweden, Finland and Norway. Overall, the majority of Type 2 diabetic patients (80%) were treated by haemodialysis, 14% by peritoneal dialysis, and 6% had a functioning renal transplant. However, transplantation was the preferred option in young patients (48% of 25-34 year olds) and in Sweden and Norway (45% of all Type 2 patients). PMID- 8243862 TI - Current status of type 2 (non-insulin-dependent) diabetic subjects on dialysis therapy in Japan. AB - According to a national survey of dialysis patients in Japan conducted by the Japanese Society for Dialysis Therapy, there were 1,033 patients on dialysis in the Shiga area which has a population of about 1.2 million. Of these 1,033 dialysis patients 140 were the result of diabetic nephropathy. From four hospitals affiliated to Shiga University of Medical Science the medical records of 90 diabetic subjects on dialysis therapy were reviewed and various clinical parameters were analysed and compared with those of patients with chronic glomerulonephritis. Since only one patient had Type 1 (insulin-dependent) diabetes, the remaining 89 with Type 2 (non-insulin-dependent) diabetes were used for this study. The significantly different variables between patients with Type 2 diabetes and chronic glomerulonephritis were age (60.4 vs 54.6 years, p < 0.05), BMI (22.4 vs 20.6 kg/m2, p < 0.001), cardiothoracic ratio (56.4 vs 53.3%, p < 0.001), mean blood pressure (110 vs 117 mmHg, p < 0.05), serum creatinine (9.0 vs 11.5 mg/dl, p < 0.001), serum urea-N (98.2 vs 115.5 mg/dl, p < 0.001), serum total protein (6.0 vs 6.5 g/dl, p < 0.001) and serum albumin (3.5 vs. 3.9 g/dl, p < 0.001). Serum levels of cholesterol and triglyceride were not significantly different between two groups, though the prevalence of electrocardiogram abnormalities, oedema, neuropathy, myocardial infarction and cerebrovascular diseases was significantly higher in the Type 2 diabetic group. These results suggested that Type 2 diabetic patients with end-stage renal disease were older, more malnourished, fluid overloaded and multi-morbid as a result of vasculopathy and neuropathy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243863 TI - Epidemiology, development and treatment of end-stage renal failure in type 2 (non insulin-dependent) diabetes mellitus. The case of mainland France and of overseas French territories. AB - The prevalence of diabetes mellitus among patients treated for end-stage renal failure by dialysis in France was studied in two stages (UREMIDIAB Study). The first stage consisted of a questionnaire which was mailed to all dialysis centres in mainland France. The response rate was 80.8%, resulting in a study population of 12,903 patients. Of these patients 884 were declared diabetic (6.9%). Later 295 of them were interviewed by seven specially-trained physicians who checked the medical records together with the nephrologist in charge. Plasma C-peptide was measured in almost all of the patients. Effectively, 1.4% were found to have Type 1 diabetes and 5.5%, Type 2. Diabetic nephropathy was found to be the only primary renal diagnosis among 93.9% of Type 1 diabetic patients and 36.8% of Type 2. Of the latter 51.6% had a non-diabetic cause of renal failure. In the second stage a survey was later conducted in 13 of 14 dialysis centres located in the remote overseas French territories. Among 934 patients 1.04% were Type 1 diabetic and 19.67% Type 2 (22.9% altogether). Type 2 diabetic patients treated overseas were essentially non-Caucasians (92.6%). The sex ratio was 0.54 in the overseas territories vs 1.4 in the mainland. We conclude that the prevalence of diabetes among people on dialysis is low in mainland France. But there are striking differences in the prevalence of Type 2 diabetes among dialysis patients in mainland France and its overseas territories. These differences are not related to access to dialysis facilities. PMID- 8243864 TI - Survival and predictors of death in dialysed diabetic patients. AB - The objective of this study was to examine diabetic patients at the time of admission to maintenance haemodialysis and to follow them for 36 months in order to define predictors of cardiovascular and non-cardiovascular death. This prospective study comprised all consecutive diabetic patients admitted to 28 German dialysis centres between January 1985 and October 1987; 196 patients were examined, 67 Type 1 (insulin-dependent) diabetic (43 male, 24 female; median age 49 years, range 22-73) and 129 Type 2 (non-insulin-dependent) diabetic patients (54 male, 75 female; 64 years, range 37-82). Outcome measures were death, i.e. myocardial infarction, sudden death, cardiac death of other causes, stroke and non-cardiovascular death. Actuarial survival 36 months after the beginning of dialysis was similar in Type 1 (40%) and Type 2 diabetic patients (43%) despite the age difference. Causes of death were myocardial infarction (18%), sudden death (18%), other cardiac causes (18%); stroke (6%); septicaemia (17%) mostly originating from diabetic foot problems; and interruption of therapy. Survival rates and the proportion dying from cardiac causes were similar in patients with diabetic nephropathy or with other primary chronic renal disease and coincidental diabetes. On dialysis, de novo amaurosis or de novo amputation was not observed in any patient. The strongest predictor of myocardial infarction or sudden death was serum lipids on admission. Duration of hypertension, blood pressure at the time of admission to dialysis, left ventricular hypertrophy or end-diastolic diameter by echocardiography, Sokolow index and average predialysis blood pressure, smoking, interdialytic weight gain and type of dialysis were not predictive of cardiovascular death or death by all causes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243865 TI - A review of the recent epidemiological data on the worldwide incidence of type 1 (insulin-dependent) diabetes mellitus. World Health Organization DIAMOND Project Group. AB - Nearly 70 registries from more than 40 countries have collected and published incidence data of childhood Type 1 (insulin-dependent) diabetes mellitus up to the end of the 1980s. The majority of incidence data comes from regions of high incidence i.e. from Europe and North America. All these published data facilitate the descriptive comparison of incidence and variation of the occurrence of Type 1 diabetes roughly throughout the northern hemisphere. The aim of this paper is to review and compare the most recent epidemiology data on the incidence of Type 1 diabetes among children under the age of 15 years. A clear difference in incidence appeared between northern and southern hemisphere with no countries below the equator having an incidence greater than 15.0 per 100,000. In contrast above the equator the disease is common. Between continents the variation in incidence showed that the lowest incidences were found in Asia, followed by Oceania (Australia and New Zealand), South and North America, and the highest rates were in Europe. The incidence varied from 0.6 per 100,000 in Korea and Mexico to 35.3 per 100,000 in Finland showing prominent worldwide variation in incidence of Type 1 diabetes. The largest intracontinental variation in incidence appeared in Europe, varying from the highest in Finland to the lowest (4.6 per 100,000) in northern Greece. The highest incidence in the world was in northern Europe, but within the continent scale there were some striking exceptions from the overall level of incidence.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243866 TI - The preventive effect of aldose reductase inhibition on diabetic optic neuropathy in the BB/W-rat. AB - A polyol-pathway-related mechanism has been invoked in the pathogenesis of murine and human diabetic peripheral neuropathy in which progressive axonal atrophy and axo-glial dysjunction constitute the cardinal structural abnormalities. We have previously reported similar neuroanatomical changes in the optic nerve of 6-month diabetic BB/W-rats. In the present study we demonstrate progression of axonal atrophy and axo-glial dysjunction in the optic nerve in 12-month diabetic BB/W rats. These structural lesions showed highly significant correlations with the associated prolongation of the latencies of the visual evoked potentials, suggesting that axo-glial dysjunction and axonal atrophy are major determinants for impaired optic nerve function. As in peripheral nerve, the polyol-pathway is present in the optic nerve and is activated by hyperglycaemia and galactosaemia. In this study we further examined the treatment effect of the aldose reductase inhibitor ponalrestat, given from 3 weeks of diabetes and continued throughout the study protocol. This regimen resulted in complete prevention of axo-glial dysjunction, and had a significant ameliorating effect on visual evoked potential latencies, but had no effect on optic nerve axonal atrophy. This latter finding differs from the effect of aldose reductase inhibition on diabetic peripheral nerve and suggests that axonal atrophy of central nerve tracts in diabetes may be the consequence of other metabolic abnormalities or alternatively the present regimen was insufficient to protect central axons from the effects of an increased activity of the polyol pathway. PMID- 8243867 TI - Involvement of non-esterified fatty acid oxidation in glucocorticoid-induced peripheral insulin resistance in vivo in rats. AB - The mechanism by which glucocorticoids induce insulin resistance was studied in normal rats administered for 2 days with corticosterone then tested by euglycaemic hyperinsulinaemic clamps. Corticosterone administration induced a slight hyperglycaemia, hyperinsulinaemia and increased non-esterified fatty acid levels. It impaired insulin-stimulated total glucose utilization (corticosterone 15.7 +/- 0.7; controls 24.6 +/- 0.8 mg.kg-1 x min-1), as well as residual hepatic glucose production (corticosterone 4.9 +/- 1.0; controls 2.0 +/- 0.7 mg.kg-1 x min-1). During the clamps, insulin did not decrease the elevated non-esterified fatty acid levels in corticosterone-administered rats (corticosterone 1.38 +/- 0.15, controls 0.22 +/- 0.04 mmol/l). Corticosterone administration decreased the in vivo insulin-stimulated glucose utilization index by individual muscles by 62 +/- 6%, and the de novo glycogen synthesis by 78 +/- 2% (n = 8-9 muscles). GLUT4 protein and mRNA levels were either unchanged or slightly increased by corticosterone administration. Inhibition of lipid oxidation by etomoxir prevented corticosterone-induced muscle but not hepatic insulin resistance. In conclusion, glucocorticoid-induced muscle insulin resistance is due to excessive non-esterified fatty acid oxidation, possibly via increased glucose fatty-acid cycle ultimately inhibiting glucose transport, or via decreased glycogen synthesis, or by a direct effect on glucose transporter translocation or activity or both. PMID- 8243868 TI - Decreased skin blood flow early in the course of streptozotocin-induced diabetes mellitus in the rat. AB - We have previously used laser Doppler technology to demonstrate that skin blood flow is reduced in Type 1 (insulin-dependent) diabetic patients. The possibility of using the skin as an extremely accessible indicator of diabetic microvascular disease is attractive. The streptozotocin diabetic rat is an appealing potential animal model. We performed measurements of skin blood flow in two rat species, nine Sprague Dawley (SD) rats and nine Wistar Kyoto (WKY) rats, observing early changes following the inception of diabetes. Four of the SD rats and five of the WKY rats were made diabetic, the rest serving as controls. There were no significant differences in skin blood flow between the two rat strains. As in man, there appear to be rat skin sites with primarily nutritive capillary supply and those with arteriovenous anastomotic predominance. The back and base of tail, both hair-covered areas, demonstrated low flow characteristics, consistent with nutritive perfusion. In contrast, the plantar surface of the paw behaved similarly to the finger or toe pulps in man, sites of arteriovenous perfusion, with high basal flow and a marked increment with thermal stimulation. In diabetic rats of both species, there was significantly lower flow at the back and base of tail than in non-diabetic animals. The differences were of the order of 30-40%. As a function of time, the decrease in blood flow at the base of tail parallelled the increase in glycohaemoglobin levels in the diabetic rats. In contrast, blood flow at the plantar surface of the paw was unchanged throughout the 3-month post streptozotocin observation period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243869 TI - The BBZ/Wor rat: clinical characteristics of the diabetic syndrome. AB - The BBZ/Wor rat is a model of obesity and autoimmune diabetes mellitus developed by crossing the BB/Wor and Zucker rats. We studied circulating glucose and insulin levels, islet morphology and lymphocyte subsets in lean and obese BBZ/Wor rats before and after the onset of diabetes, and studied the clinical course of diabetes in animals after interruption of exogenous insulin therapy. Lean BBZ/Wor rats developed insulin-dependent diabetes and died in ketoacidosis within 1 week after cessation of insulin injections. Diabetes also developed in obese rats, but these animals were not insulin-dependent and survived for months without insulin therapy. The islets of the lean diabetic rats revealed complete destruction of pancreatic beta cells and plasma insulin levels were virtually undetectable. In contrast, the islets of the obese rats revealed insulitis and substantial beta cell loss, however autoimmune beta-cell destruction was incomplete, and residual beta cells were presumably responsible for the presence of measurable levels of plasma insulin and the long-term survival of obese diabetics without insulin therapy. Obese rats were hyperinsulinaemic, developed diabetes significantly earlier, and with a greater incidence than lean rats, suggesting a possible relationship between enhanced beta-cell metabolic activity and immune destruction. Obese males became diabetic more frequently and at an earlier age than obese females and lean rats of both sexes, suggesting a role for gender in the pathogenesis of diabetes. We conclude that the BBZ/Wor rat is a unique animal model for investigating the interaction of obesity, beta-cell metabolism, autoimmune insulitis and genetic predisposition to diabetes. PMID- 8243870 TI - Vasoactive intestinal polypeptide-augmented insulin release: actions on ionic fluxes and electrical activity of mouse islets. AB - Vasoactive intestinal polypeptide is a pancreatic neurotransmitter which augments insulin release. To obtain more detailed information on its mode of action on the pancreatic beta cell we studied the effect of vasoactive intestinal polypeptide on 86Rb+ efflux, 45Ca2+ uptake, electrical activity and second messenger systems of isolated mouse islets. Vasoactive intestinal polypeptide enhanced insulin release and 45Ca2+ uptake in a concentration-dependent manner, and was effective at non-stimulatory and stimulatory glucose levels. It increased glucose-induced electrical activity but was without effect on either glucose-mediated changes of 86Rb+ efflux, cAMP or inositol-1,4,5-trisphosphate content. It is suggested that vasoactive intestinal polypeptide augments insulin release by increasing the uptake of Ca2+ into the cell by as yet undefined mechanisms. PMID- 8243871 TI - Increased aortic endothelial death and enhanced transendothelial macromolecular transport in streptozotocin-diabetic rats. AB - Hypertension, cigarette smoking and diabetes mellitus are well-known risk factors for atherosclerosis and coronary heart disease. Repeated endothelial cell injury and increased lipid entry have been suggested as initiating events in atherogenesis. Our previous studies have demonstrated that the frequency of endothelial cell death and associated endothelial permeability were significantly increased in the aorta of spontaneously hypertensive rats and chronic oral nicotine-treated rats. In the present investigation, we examined the hypothesis that diabetes also increases the frequency of arterial endothelial cell death and hence transendothelial macromolecular transport, which may have some implications in increasing lipid entry and thus accelerating atherogenesis. Diabetes was induced in 15 male Sprague-Dawley rats by intraperitoneal injection of 60 mg streptozotocin per kg body weight. The duration of diabetes was 6 weeks. A group of 15 age-matched rats, injected only with the buffer and maintained over the same time period, served as the controls. In en face preparations of the thoracic aorta, IgG-containing dead endothelial cells were identified by an indirect immunoperoxidase method, and endothelial leakage to Evans blue-albumin complexes was quantified by fluorescence microscopy. Diabetic rats, compared to control rats, had significantly higher values for the frequency of endothelial cell death (0.77 +/- 0.10% vs 0.38 +/- 0.04%; p < 0.005 by two-tailed, unpaired Student's t test) and the number density of Evans blue-albumin leaky foci (4.33 +/- 0.48/mm2 vs 2.99 +/- 0.38/mm2; p < 0.05 by two-tailed, unpaired t-test) in the aorta.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243872 TI - Intranasal glucagon treatment relieves hypoglycaemia in children with type 1 (insulin-dependent) diabetes mellitus. AB - The aim of the present study was to compare intra-nasal glucagon with subcutaneous glucagon as a treatment of insulin-induced hypoglycaemia in 11 children, 7-12 years old, with Type 1 (insulin-dependent) diabetes mellitus. Hypoglycaemia (1.6 +/- 0.1 vs 1.8 +/- 0.2 mmol/l) was induced twice in each child by continuous insulin and variable glucose infusions. One milligram of intranasal glucagon or 0.5 mg of subcutaneous glucagon was given in a randomized order. At 15 min after the administrations of either intranasal or subcutaneous glucagon, the blood glucose concentration increased by 1.5 +/- 0.2 mmol/l or 1.7 +/- 0.2 mmol/l above the glucose nadir, respectively. After nasal administration, the maximal rise in blood glucose was seen after 25 min. Subcutaneous injections induced higher and more sustained plasma glucagon concentrations but the children suffered more often from nausea than when they were treated intranasally. In conclusion, treatment with intranasal glucagon seems to be efficient and results in a rapid correction of insulin-induced hypoglycaemia with few side-effects. PMID- 8243873 TI - Differential effects of type 2 (non-insulin-dependent) diabetes mellitus on pentosidine formation in skin and glomerular basement membrane. AB - Pentosidine is an advanced Maillard/glycation reaction product the formation of which in human skin is significantly increased in Type 1 (insulin-dependent) diabetes mellitus and correlates with the severity of diabetic complications. Preliminary data in a limited number of Type 2 (non-insulin-dependent) diabetic individuals showed that skin pentosidine was not significantly elevated, raising the question of whether statistical power was insufficient for differences to be revealed, or whether pentosidine did not form because biological factors intrinsic to Type 2 diabetes affected the advanced Maillard reaction altogether. To resolve this question, pentosidine levels were measured in 209 human skin samples obtained at autopsy and in purified glomerular basement membranes from 45 subjects of various ages, with and without Type 1 and Type 2 diabetes and uraemia. Pentosidine increased exponentially in skin but curvilinearly in glomerular basement membranes, and reached 75 and 50 pmol/mg collagen at projected 100 years, respectively. Skin levels were not significantly elevated in individuals with Type 2 diabetes (p > 0.05). In contrast, pentosidine levels in glomerular basement membranes were elevated above the 95% confidence interval in the majority of diabetic patients regardless of the type of diabetes and in all individuals on haemodialysis. These data clearly demonstrate that the advanced Maillard reaction is indeed accelerated in Type 2 diabetes and strongly suggest that differences in pentosidine accumulation rates may be due to differences in collagen turnover. In diabetes and uraemia, accelerated Maillard reaction mediated protein crosslinking, as reflected by pentosidine, may contribute to decreased turnover of the extracellular matrix, sclerosis and thickening of basement membranes. PMID- 8243874 TI - Insulin excess counteracts the effects of HDL on intracellular sterol accumulation in cultured human skin fibroblasts. AB - Both Type 1 (insulin-dependent) and Type 2 (non-insulin-dependent) diabetic individuals are at increased risk of developing ischaemic heart disease. Insulin excess, present in both diabetic groups, may play an important pathophysiologic role in accelerating the atherogenic process. In this study, cultured human skin fibroblasts were incubated with varying concentrations of insulin to test the role of insulin on cell cholesterol homeostasis and on HDL3-mediated removal of excess cholesterol from cells. Insulin excess (1-100 nmol/l) resulted in a significant dose-dependent reduction in HDL3-mediated cholesterol efflux from the intracellular unesterified cholesterol pool of cultured human skin fibroblasts. Similar insulin concentrations resulted in impaired HDL-mediated cholesterol efflux from the cell membrane but had no effect on non-HDL-mediated efflux. The effect of insulin on cholesterol esterification and biosynthesis was assessed by 14C-oleate labelling. The addition of HDL3 (50 micrograms) resulted in a significant decrease in 14C-labelled cholesterol ester, reflecting a decrease in intracellular unesterified cholesterol, which was partially reversed by the addition of insulin. Insulin had no effect on the incorporation of 14C-oleate into unesterified cholesterol. During simultaneous incubation of fibroblasts with LDL and HDL, insulin resulted in an increase in cholesterol esterification and inhibited ability of HDL to promote the decrease in esterification. Thus, we have shown that insulin excess counteracts the beneficial effects of HDL that involve removal of cellular cholesterol and may in part promote atherogenesis by this mechanism. PMID- 8243875 TI - Relationships between dyspeptic symptoms and gastrointestinal motility in patients with type 1 (insulin-dependent) diabetes mellitus. AB - Reports on motor abnormalities in Type 1 (insulin-dependent) diabetes mellitus are inconsistent. In 20 Type 1 diabetic patients and in 11 control subjects antroduodenojejunal manometry was performed under euglycaemic conditions in order to examine the prevalence of gastric and small intestinal motor abnormalities in relation to dyspeptic symptoms and the degree of cardiac autonomic neuropathy. In diabetic patients compared to control subjects phase III (regular, high-amplitude contractile activity at maximal frequency) involved the gastric antrum less often (12 vs 35%, p < 0.05), the duration of phase I (motor quiescence) was shorter (6 +/- 1 vs 21 +/- 4 min, p < 0.002) and in phase II (irregular motor activity) the frequency of duodenal and jejunal contractions was higher. After a meal the duration of the fed state was shorter in diabetic patients with symptoms during the study than in diabetic patients without symptoms and than in control subjects (57 +/- 27 vs 157 +/- 11 and 140 +/- 13 min, p < 0.02). Postprandial antral hypomotility was seen in diabetic patients with symptoms only in the first 30 min after the meal. One hour after the meal the frequency of duodenal and jejunal contractions was again higher in diabetic patients. In diabetic patients compared to control subjects more burst activity (clusters of non-propagated high amplitude contractile activity at maximal frequency) was seen (7.9 +/- 1.6 vs 0.8 +/- 0.5% of the total time of study, p < 0.002). No correlation was found between manometric parameters and the degree of cardiac autonomic neuropathy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243876 TI - Oxygen diffusive conductance in placentae from control and diabetic women. AB - Random tissue sections of placentae from control and diabetic deliveries were analysed stereologically. The aim was to test whether or not adaptations in oxygen diffusive conductances occur to help compensate for fetal hypoxic stress in utero. Organs were from 34 control and 55 diabetic deliveries (39 from White's classes A, B, C and 16 from classes D, F/R) arranged in two major groups (ABC and DFR). Tissue samples were fixed in formalin and processed for wax histology. Stereological and physicochemical quantities were used to calculate the partial conductances of six tissue layers, viz. maternal erythrocytes and plasma, villous trophoblast, villous stroma, fetal plasma and fetal erythrocytes. From partial conductances and birthweights, total and specific conductances for each placenta were determined. No differences were detected between diabetic placentae in different classes (A, B, C, D, F/R). However, both the ABC and DFR groups of diabetic placentae had higher total diffusive conductances than control organs. The increase was in the order of 7-25% and persisted even when adjustments were made for apparent differences in birthweights. The principal contributors to these changes were the post-trophoblastic (fetally located) tissue layers. These findings suggest that the diabetic placenta adapts to facilitate the diffusion of oxygen across the placenta and, thereby, to assist the hypoxic fetus. PMID- 8243877 TI - Elevated long-term glycated haemoglobin precedes proliferative retinopathy and nephropathy in type 1 (insulin-dependent) diabetic patients. AB - The importance of glycaemic control for the development of proliferative retinopathy and nephropathy was assessed by monitoring glycated haemoglobin for 5 years or more before the diagnosis of these complications. The study comprised Type 1 (insulin-dependent) diabetic patients diagnosed at an age less than 31 years, and with diabetes duration 25 years or less. They were followed for an average of 7.9 years with 3.3 measurements per year. Of 172 patients screened for retinopathy 60 had no retinopathy, 104 had background retinopathy, and 8 had proliferative retinopathy The mean HbAlc (95% confidence intervals) of the groups was 6.4% (6.2-6.7%), 7.3% (7.1-7.5%) and 8.9% (8.1-9.6%), respectively (p < 0.0001); the mean duration of diabetes was 12, 18, and 17 years. Of 186 patients 7 had nephropathy (albuminuria > 200 mg/l). Mean HbAlc in patients without nephropathy was 7.0% (6.8-7.1%) and in patients with nephropathy 8.8% (7.8-9.9%, p < 0.001). Mean diabetes duration was 16 years in both groups. Multiple logistic regression including mean HbAlc, age at onset, duration, sex, and hypertension, was for both proliferative retinopathy and nephropathy significant only for mean HbAlc. In all cases, proliferative retinopathy and nephropathy were preceded by poor glycaemic control over several years, suggesting that these complications are caused by poor glycaemic control. PMID- 8243878 TI - Effects of recombinant human insulin-like growth factor I and insulin on counterregulation during acute plasma glucose decrements in normal and type 2 (non-insulin-dependent) diabetic subjects. AB - Insulin-like growth factor I (65 micrograms/kg) or insulin (0.1 IU/kg) were injected i.v. on two separate occasions in random order in normal and in Type 2 (non-insulin-dependent) diabetic subjects. Insulin-like growth factor I and insulin injection resulted in identical decrements of plasma glucose concentrations after 30 min but in delayed recovery after insulin-like growth factor I as compared to insulin in both groups (p < 0.05 insulin-like growth factor I vs insulin). Counterregulatory increases in plasma glucagon, adrenaline, cortisol and growth hormone concentrations after hypoglycaemia (1.9 +/- 0.2 mmol/l) in normal subjects were blunted after insulin-like growth factor I administration compared to insulin (p < 0.05). Plasma glucose in Type 2 diabetic subjects did not reach hypoglycaemic levels but the acute glucose decrease to 4.5 +/- 0.8 mmol/l was associated with significantly lower responses of plasma glucagon and adrenaline but higher cortisol levels after insulin-like growth factor I compared to insulin (p < 0.003). Plasma concentrations of non-esterified fatty acids and leucine decreased similarly after insulin-like growth factor I and insulin in both groups. The present results demonstrate that insulin-like growth factor I is capable of mimicking the acute effects of insulin on metabolic substrates (plasma glucose, non-esterified fatty acids, leucine). The decreases of plasma glucose were similar after both peptides in normal and in diabetic subjects who were presumably insulin resistant. Counterregulatory hormone responses to plasma glucose decrements differed, however, between insulin-like growth factor I and insulin and in the diabetic and the control subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243879 TI - Diabetes control and pulmonary function in type 1 (insulin-dependent) diabetes mellitus. PMID- 8243880 TI - Are patients with fibrocalculous pancreatic diabetes a subset of chronic calcific pancreatitis of the tropics with genetic predisposition to type 1 (insulin dependent) diabetes? PMID- 8243881 TI - Fetal growth and impaired glucose tolerance in men and women. PMID- 8243882 TI - Problems related to definitions and epidemiology of type 2 (non-insulin dependent) diabetes mellitus: studies throughout the world. AB - Many studies of Type 2 (non-insulin-dependent) diabetes mellitus assume that the condition is homogeneous and clearly defined. There are, however, several problems with these assumptions. Thus, definition of Type 2 diabetes is one of exclusion of other types (insulin-dependent, malnutrition-related, gestational and other rarer types) and inevitably contains a heterogeneous group of disorders the aetiology of which is largely unclear, and separation from the insulin dependent type can be problematic. Diagnosis is also imprecise in asymptomatic subjects due to the lack of accurate diagnostic tools and lack of clear distinction of impaired glucose tolerance. An alternative to the oral glucose tolerance test is urgently needed. Epidemiological studies of Type 2 diabetes and its complications are also fraught with difficulties due to variability of the oral glucose tolerance test, potential problems in glucose measurement, heterogeneity, population selection and problems in international comparisons due to differing age structures and life expectancy. Great care is needed in all studies of Type 2 diabetes to ensure that the groups under study are properly selected, well-defined and fully described. PMID- 8243883 TI - Renal function changes in middle-aged and elderly Caucasian type 2 (non-insulin dependent) diabetic patients--a review. PMID- 8243884 TI - Storage temperature and differing methods of sample preparation in the measurement of urinary albumin. AB - Microalbuminuria is a predictor of persistent proteinuria, renal failure and cardiovascular disease and therefore accurate determination of urinary albumin concentration is important. We examined the stability of albumin in urine under different conditions of storage, temperature and sample preparation. There was no significant difference in urinary albumin concentration between fresh urine and urine stored at either 4 degrees C or 20 degrees C for up to 7 days. Similarly in urine samples from diabetic patients there was no significant difference in albumin concentration at levels ranging from 1.3 to 1999.3 mg/l between fresh urine at 4 degrees C and urine stored frozen for 1 week, 1 month or 6 months. Neither storage temperature (-20 degrees C or -40 degrees C) nor centrifugation of sample prior to assay made a significant difference to the albumin concentration. Multiple freezing and thawing of urine samples during 6 weeks of storage at -20 degrees C made no difference to albumin concentrations. Storage of urine samples in either polypropylene, polystyrene or borosilicate glass tubes did not result in a significant change in urinary albumin concentration after either 1 week or 1 month at -20 degrees C although, after 1 month of storage, urinary albumin concentrations tended to be lower by an average of approximately 7%. In tubes to which gelatine had been added this was reduced to 4%. We conclude that fresh urine can be kept at 4 degrees C or 20 degrees C for up to 7 days. Frozen urine samples can be stored for up to 6 months before assay without any loss of albumin concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243886 TI - Abstracts of the 29th Annual Meeting of the European Association for the Study of Diabetes. Istanbul, Turkey, 6-9 September 1993. PMID- 8243885 TI - Pre-diabetic blood pressure predicts urinary albumin excretion after the onset of type 2 (non-insulin-dependent) diabetes mellitus in Pima Indians. AB - Blood pressure was measured in 490 non-proteinuric Pima Indians from the Gila River Indian Community in Arizona at least 1 year before the diagnosis of Type 2 (non-insulin-dependent) diabetes mellitus. Urine albumin concentration was measured in the same subjects 0-24 years (mean 5 years) after diabetes was diagnosed. Prevalence rates of abnormal albumin excretion (albumin-to-creatinine ratio > or = 100 mg/g) after the onset of Type 2 diabetes were 9%, 16%, and 23%, respectively, for the lowest to highest tertiles of pre-diabetic mean blood pressure. When controlled for age, sex, duration of diabetes and pre-diabetic 2-h post-load plasma glucose concentration, higher pre-diabetic mean blood pressure predicted abnormal urinary excretion of albumin after the onset of diabetes. This finding suggests that the higher blood pressure seen in diabetic nephropathy is not entirely a result of the renal disease, but may precede and contribute to it. PMID- 8243887 TI - Wilms' tumor (WT1) gene expression in rat decidual differentiation. AB - The Wilm's tumor suppressor gene (WT1) encodes a zinc-finger containing transcription factor that is selectively expressed in the developing urogenital tract, where it is thought to play a role in the differentiation of these tissues. We have used immunocytochemistry and in situ hybridization to study WT1 expression in the rat uterus during normal development and pregnancy from 0 to 20 days post coitum (p.c.). WT1 mRNA was abundant in uterine stroma from juvenile rats, but was much less abundant in uterine tissue from sexually mature rats; WT1 expression is not affected by ovariectomy or by treatment with estradiol or estradiol plus progesterone. WT1 gene was highly expressed, however, in the endometrial cells of early pregnancy. On day 6 p.c. WT1 mRNA was detected in anti mesometrial decidual cells, and WT1 immunoreactivity was concentrated in the nuclei of these cells. All cells of fully-developed deciduoma at 7-8 days p.c. demonstrated WT1 expression. WT1 was not detected in trophoblast/placental tissues but remained abundant in the decidua basalis until parturition. The expression of WT1 was compared with insulin-like growth factor-II (IGF-II) and its receptor in the decidual since it has been shown that IGF-II gene transcription is repressed by WT1 in vitro. However, no spatiotemporal correlation in the expression of these three genes was found in differentiation of the rat decidua. In summary, these data suggest a role for WT1 in decidualization, since its expression is activated during the differentiation of uterine stromal cells into decidual cells. PMID- 8243888 TI - Cooperation between retinoic acid and phorbol esters enhances human teratocarcinoma differentiation. AB - This study explored cooperation between the retinoic acid (RA) and protein kinase C (PKC) pathways during differentiation of the multipotential human teratocarcinoma (TC) cell line NTERA-2 clone D1 (abbreviated NT2/D1). We report here that, compared to RA treatment alone, RA combined with the PKC stimulator 12 O-tetradecanoylphorbol-13-acetate (TPA) enhanced the regulated expression of the immunophenotypic differentiation markers SSEA-3, a globo-series carbohydrate, and the ganglio-series carbohydrate antigens GD2 and GD3. Northern analysis and transient transfection assays revealed that TPA co-treatment augmented the RA induced expression and activation of the RA nuclear receptor-beta (RAR-beta), one early marker of RA response in NT2/D1 cells. This finding was extended with transient co-transfection experiments using a PKC-alpha expression vector which revealed that the PKC pathway can augment the activation of RAR-beta by RA. These experiments establish PKC as a modulator of RAR-beta expression in NT2/D1 cells. Similarly, experiments showed that RA can modulate activation of the PKC responsive AP-1 complex, a transcription factor rapidly activated by TPA. Northern analysis and transient transfection assays revealed that, compared to TPA treatment alone, RA and TPA augmented the expression and transcriptional activity of AP-1 in NT2/D1 cells. In contrast, transient transfection assays revealed no cooperative effect between RA and TPA in HeLa cells, indicating that this effect in NT2/D1 cells is cell type-specific. In summary, these studies show that stimulation of the PKC second messenger pathway can modulate tumor differentiation and transcriptional activation of a retinoid receptor associated with RA response. PMID- 8243889 TI - 9-cis and all-trans retinoic acid induce a similar phenotype in human teratocarcinoma cells. AB - Prior work has shown that all-trans retinoic acid (t-RA) treatment of the human teratocarcinoma (TC) cell line NTERA-2 clone D1 (abbreviated NT2/D1) induces a neuronal phenotype and other cell lineages. This study sought to explore the potential of 9-cis retinoic acid (9-cis RA) as a differentiation-inducing agent of this multipotent cell. Findings reported here show that 9-cis RA induced a phenotype similar to t-RA treatment of NT2/D1 cells. This similarity extended to their effects on the nuclear receptors retinoic acid receptor-beta (RAR-beta) and retinoid X receptor-alpha (RXR-alpha). Both retinoids prominently augmented RAR beta expression and transactivated a reporter plasmid containing putative RAR response elements (RAREs) with direct repeats separated by five nucleotides (DR5). Both retinoids had no appreciable effect on RXR-alpha expression and both minimally transactivated a reporter plasmid containing putative RXR response elements (RXREs) with direct repeats separated by one nucleotide (DR1). These studies suggest that 9-cis RA and t-RA activate common events during retinoid mediated NT2/D1 differentiation. This hypothesis was supported by the finding that NT2/D1 cells rendered refractory to t-RA (NT2/D1-R1) were also resistant to 9-cis RA. To discover alterations that could confer retinoid-refractoriness, retinoid receptor expression was examined in NT2/D1-R1 cells. In contrast to NT2/D1, the NT2/D1-R1 cell was found to have reduced RXR-alpha expression at the level of total cellular RNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8243890 TI - Vacuolar apical compartment (VAC) in breast carcinoma cell lines (MCF-7 and T47D): failure of the cell-cell regulated exocytosis mechanism of apical membrane. AB - We have previously shown that an integral plasma membrane glycoprotein (AP2) is highly polarized to the apical domain in confluent Madin-Darby canine kidney (MDCK) epithelial cells. However, when the monolayers are prevented from forming intercellular contacts, approximately 60% of the AP2 cellular content is stored in the intracellular vacuolar apical compartment (VAC). In the current work we found that AP2 was present in the non-tumorigenic human mammary epithelial cell line MCF-10A, in the breast carcinoma cell lines MCF-7 and T47D, and in breast ductal carcinomas in vivo. By radioimmunoassay, an intracellular compartment of AP2 was identified in the mammary cell lines in culture. In MCF-10A, this compartment behaved as in MDCK cells; namely it was observed only when the cells cannot form cell-cell contacts. However, in the carcinoma cell lines MCF-7 and T47D, a significant AP2 intracellular compartment was observed also under conditions permissive for the formation of intercellular contacts. These results were confirmed by immunofluorescence and immunoelectron microscopy experiments that showed VACs in MCF-7 and T47D, even in cells with extensive intercellular contacts. In MCF-7 cells, the addition of serum caused a partial decrease of the AP2 intracellular compartment. The exocytosis of VACs occurred towards the center of multi-cellular groups, forming intercellular lumens, similar to those transiently observed in MDCK cells and to structures described by others during embryo development. Altogether, these results suggest that VAC exocytosis is controlled by cell-cell contact signalling, which may be defective in carcinoma cells. PMID- 8243891 TI - Characterization of growth and differentiation of normal human submandibular gland epithelial cells in a serum-free medium. AB - Parenchymal tissue of human submandibular glands was cultured in a serum-free medium consisting of a 1:9 mixture of Dulbecco's modified Eagle's medium and MCDB 153 supplemented with 10 ng/ml epidermal growth factor, 10 microM dexamethasone and 1 microgram/ml insulin. Cultivation of the tissue in this medium resulted in propagation of loosely arranged epithelioid cells on plastic, without the necessity of a matrix. Epidermal growth factor significantly enhanced mitogenesis of cultured cells, which expressed specific high- and low-affinity receptors for epidermal growth factor. The epithelioid cells were found to represent the undifferentiated ultrastructure of ductal cells. Immunocytochemically, cultured epithelioid cells expressed antigens specific to basal cells of the intra- and interlobular ducts in situ, including cytokeratins 3 and 6 and cytokeratins 13 and 16, vimentin, and alpha-smooth muscle actin. Moreover, cytoplasm of the cells was immunostained using antibody against the basement membrane component, type IV collagen. These results suggested that cultured epithelioid cells are undifferentiated ductal cells, which have the characteristics of basal cells of the intra- and/or interlobular ducts. Cultured epithelioid cells maintained the characteristics for serial passage until the time that the cultures were confluent. On the other hand, several stratified foci developed on the confluent monolayer. The stratified cells were strongly positive for cytokeratins 3 and 6, but negative for vimentin, alpha-smooth muscle actin and type IV collagen. Moreover, the stratified cells were strongly stained with the antibody against epithelial membrane antigen. This antibody stained the luminal membrane domain of salivary epithelial cells. Electron micrograph of the vertical section through the foci revealed stratified cell layers with a gradual transition from basal cells to squamous epidermoid cells. This result suggests that cultured epithelioid cells, which have the characteristics of basal cells of the intra- and/or interlobular duct, have the potential to differentiate into luminal duct cells. PMID- 8243892 TI - Appearance and distribution of entactin in the early chick embryo. AB - Entactin is a sulfated glycoprotein of basement membranes and recent data indicate that it may play a major role in extracellular matrix (ECM) assembly and in modulating the activities of the other molecular components. We investigated the time of appearance and subsequent distribution of entactin during the earliest stages of morphogenesis and its involvement in the first major cellular migrations and interactions in the chick embryo. Entactin is first detected in the epiblast and in the hypoblast at the blastula stage. The accumulating ECM displays intense presence of entactin in the space between the epiblast and the hypoblast at late blastula. Entactin is increasingly abundant in the neural plate and in the ECM and also at least transiently in many mesodermal tissues such as the notochord, the developing heart and somites in the early chick embryo. Immunogold labeling revealed a punctate pattern of entactin distribution in the ECM during the gastrula, neurula and at later stages and at all levels within the embryo. Because of its early appearance in more than one germ layer, entactin may be important in the formation of most embryonic structures. Entactin is detected at the same developmental time and co-localizes with laminin. Antibodies to entactin do not interfere with triggering of the first major cell movements but perturb directional migration of these cells. It would seem that entactin plays a functional role in the directed migration of cells and does not seem to affect cell adhesion during the period of the first morphogenetic events in the early chick embryo. PMID- 8243893 TI - Pair-rule gene expression in the somitic stage chick embryo: association with somite segmentation and border formation. AB - In vertebrates, metameric organization is high-lighted by the formation of somites from mesenchymal cells of the segmental plate which then differentiate into dermamyotomal and sclerotomal tissues. The resegmentation of the sclerotome into rostral and caudal halves follows, coincident with the production of specific extracellular matrix molecules at the abutment of these two cell types. Ultimately, cells from the caudal sclerotome migrate ventrally and contribute to the chondrogenic prevertebrae. The objective of this work is to investigate the molecular steps regulating these events. Our study is focused on the paired-box containing genes, which have been implicated in delineating boundaries early in development. A chick embryo system, which is readily accessible to manipulation and observation during early development, is used in this study. We have identified the existence of the paired-box motif in the chicken genome by polymerase chain reaction and hybridization with the mouse Pax 1 paired-box sequence. Expression of paired-box genes occurs early in development as shown by Northern analysis, and is localized by in situ hybridization to the edge of each somite, a patch at the central core of each somite, and the periphery of the neural tube. This specific spatial pattern of expression is consistent with the hypothesis that the pair-rule genes function as effecters of border formation in the early embryo. Moreover, the patch of positive cells at the center of a resegmenting somite appear to migrate ventrally, and may contribute to structures of the prevertebrae. These findings are relevant to our understanding of the mechanism of somite resegmentation and implicate the involvement of pair-rule genes in the process. PMID- 8243894 TI - Peroxisomes and peroxisomal enzymes along the crypt-villus axis of the rat intestine. AB - The development of peroxisomes and expression of their enzymes were investigated in differentiating intestinal epithelial cells during their migration along the crypt-villus axis. Sequential cell populations harvested by a low-temperature method were identified by microscopy, determination of alkaline phosphatase and sucrase activities and incorporation of [3H]-thymidine into DNA. Ultrastructural cytochemistry after staining for catalase activity, revealed the presence of peroxisomes in undifferentiated stem cells located in the crypt region. Morphometry indicated that the number of these organelles increased as intestinal epithelial cells differentiate. Catalase activity was higher in the crypt cells than in the mature enterocytes harvested from villus tips. On the other hand, an increasing gradient of activity was observed from crypts to villus tips for peroxisomal oxidases, i.e. fatty acyl coA oxidase, D-amino acid oxidase and polyamine oxidase. These findings indicate that biogenesis of peroxisomes occurs during migration of intestinal epithelial cells along the crypt-villus axis and that peroxisomal oxidases contribute substantially to the biochemical maturation of enterocytes. PMID- 8243895 TI - Child health news in a metropolitan newspaper. PMID- 8243896 TI - Qualitative research on career choices. PMID- 8243897 TI - Timing of data collection. PMID- 8243898 TI - Family medicine leadership: developing an American primary care research agenda. PMID- 8243899 TI - Special series: hospital privileges for family physicians. PMID- 8243900 TI - Letters from the front: testimony on behalf of OB privileges for family physicians. PMID- 8243901 TI - Fair hearing testimony. AB - Realizing the competitive environment of medicine, the gold standard for credentials must remain training, experience, ability, and/or competence. These two doctors have met this standard. Family physicians have cesarean privileges in Memphis, Tenn.; Wichita, Kan.; Kansas City, Mo.; Rochester, Minn.; and numerous other cities across our state and nation. With access to perinatal care of critical importance, all physicians must practice medicine to their fullest capabilities. This hospital will greatly benefit from its support of fair privileging for family physicians. And the AAFP will vigorously defend each family physician's right to practice obstetrics on both a state and national level. PMID- 8243902 TI - Family practice privileges: an update. PMID- 8243903 TI - A prospective review system of nonurgent consultation requests in a family medicine residency practice. AB - BACKGROUND: This study describes a system for prospectively reviewing non-urgent consultation or referral requests in a family practice residency program. METHODS: A committee composed of four rotating family physicians met weekly during a 22-month period to review all non-urgent proposed consultations. The committee either approved consultations or made other recommendations to the primary care physician for clinical management. Data were prospectively recorded to determine what types of referrals were most commonly made and approved, and if the number of consultations changed after the system was instituted. RESULTS: The committee reviewed 930 non-urgent consultation requests. The consultation request rate was 2.5 per 100 patients seen. The number of consultation requests declined significantly over the study period. (P = .0007) Seventy-two percent of consultation requests were approved; 28% were not and resulted in recommendations for alternate management. Non-approval was most likely for referrals to specialists in podiatry, rheumatology, endocrinology, neurology, gastroenterology, cardiology, and allergy. The approval rate for consultations was not significantly related to patient age, sex, or type of health insurance. Patients with multiple consultation requests were more likely to have a prepaid health insurance plan or governmental insurance than private insurance. CONCLUSIONS: Prospective review of consultations and referrals can result in a decrease in the rate of consultations and referrals made by family physicians. Alternative management can frequently be recommended. Potential benefits of this system may include significant medical care cost savings and education for physicians. PMID- 8243904 TI - A comparison of university and community-based family practice physician educators. AB - BACKGROUND: Differences in satisfaction with training and perceived preparation for practice have been observed among residents trained in community and university family practice residency programs. Our study was undertaken to determine the degree to which faculty training, experience, or practice patterns may contribute to these different resident experiences. METHODS: Questionnaires addressing prior training, experience, current practice content, motivation for entering academic medicine, and satisfaction with current teaching position were sent to full-time physician faculty at 60 family practice residency programs. RESULTS: The 208 faculty responses showed no differences in faculty at various types of residency programs with respect to demographic factors, prior training, or past experience. Only small differences were found in the proportion of time devoted to most professional activities or in the content of clinical practice. CONCLUSIONS: Few differences were found between faculty at university- and community-based family practice residency programs. This implies that differences in residents' perceptions of training may be due to other factors, such as institutional philosophy or self-selection. However, qualitative differences in family practice faculty activities that were not measured in this research cannot be ruled out as factors influencing residents' perceptions of their training experiences. PMID- 8243905 TI - The structure and regulation of family practice residency laboratories. AB - BACKGROUND: Increasing regulation of physician office laboratories (POLs), increased complexity of outpatient care, and technological advances have increased the need to train family practice residents in POL management. There is little information on how residency laboratories are organized, how they are regulated, and how often quality controls and proficiency testing are done. METHODS: A questionnaire was sent to 385 ACGME-approved family practice residencies to determine the scope of testing, quality-assurance activities, and administrative structure of the residency laboratory. RESULTS: Two hundred and ninety-seven programs responded, for a response rate of 77%; 91.6% were performing routine office tests and 40.7% were performing automated testing. Of programs that performed automated tests, 71.2% were performing daily normal and abnormal quality-control specimens. Proficiency testing was only performed by 57.8% of programs. The program's parent hospital viewed the residency laboratory as a POL in 55.6% of cases and as a satellite of the hospital laboratory in 32.6%. CONCLUSIONS: Residency laboratories are more frequently organized as POLs than satellite laboratories of the parent hospitals. The POL model offers an excellent opportunity to teach residents about office laboratory management. Residency laboratories should be implementing proficiency testing for both patient safety and resident teaching. Many residency laboratories will need to increase the frequency of quality controls as the new CLIA '88 regulations are implemented. Accreditation by a private organization may be an attractive option for family practice office laboratories. PMID- 8243906 TI - Improving rural health: HeartSmart--an opportunity for family physician community involvement. AB - BACKGROUND: Cardiovascular disease is a major cause of morbidity and mortality among adults in the United States. Evidence shows that risk factors for cardiovascular disease appearing during childhood are reliable predictors of cardiovascular illness in adulthood. An educational program to alter risk factors present in children is needed to help prevent future disease. This paper describes HeartSmart, a cross-age teaching program that was developed by a family physician in a rural location. The purpose of the program was to improve cardiovascular health knowledge of sixth-grade students. METHODS: Under supervision of the family physician, the local hospital's cardiac rehabilitation unit taught cardiovascular disease prevention principles to a group of high school seniors, who then developed and taught a cardiovascular disease prevention program to sixth-grade students. Pre- and post-testing of the sixth grader's cardiovascular health knowledge was conducted. RESULTS: Sixth-grade students demonstrated improved knowledge of cardiovascular fitness principles following the program. CONCLUSIONS: HeartSmart exemplifies a program developed by a family physician that can be instituted in many communities to educate children about cardiovascular fitness and disease prevention principles. PMID- 8243907 TI - Literary images of menopause. AB - In contemporary North American cultures, menopause is primarily thought of as a deficiency disease. In literature, however, authors often portray menopause differently. In various literary portrayals, menopause evokes themes of the urgency of remaining life span, the occurrence of a significant life event, and a rite of passage. The use of literature to understand the way women experience life may help deepen and enlarge the perceptions of students and residents regarding the menopause. PMID- 8243908 TI - The meaning of death to adolescents in an American Indian community. AB - BACKGROUND: A high rate of premature death exists among young Native Americans in North America. To understand the qualitative effect of this phenomenon, we undertook this study to explore the meaning of death to adolescents in a Salish American Indian community. METHODS: Standard methods of ethnography were employed: community entry, open-ended in-depth interviews using key informant sampling, audiotape and field note transcription, review of field notes for key themes, and community feedback. Interviews were conducted with seven elders and 21 adolescents in a Pacific Northwestern American Indian community selected by key informants. Probe questions and narrative accounts primarily focused on personal experience with premature death among family and peers. RESULTS: The primary themes in the study were the subjects' personal exposure to death, alcohol and drugs. Spirit Sickness (a culturally defined illness experience), and healing. CONCLUSIONS: There are persistent beliefs in Spirit Sickness among adolescents and young adults in the Salish Indian community. Personal exposure to death is a precipitant of this potentially fatal illness experience. Clinicians working with Salish Native Americans should recognize potential beliefs in this illness experience among the youths. PMID- 8243909 TI - Can effective parent education occur during emergency room visits? AB - BACKGROUND: Parents might not feel the need to make as many visits to an emergency room (ER) for a sick child if they had more information about common illnesses and their management. This study measured the effect on future ER use of parent education on common childhood illnesses. METHODS: Children who were over 6 months old and had visited the ER at least twice in the preceding year were enrolled in an experimental group (n = 118) or in a control group (n = 128) over consecutive two-month periods. The intervention consisted of a pamphlet and a videotaped presentation that discussed the features and management of common childhood illnesses. RESULTS: Over the following year, an average of 0.43 (SD = 0.9) ER visits were made by experimental subjects compared to 0.52 (SD = 1.31) by control subjects (P = .30). Twenty percent (SD = .4) and 22% (SD = .4) of subsequent illnesses of experimental and control children respectively resulted in an ER visit. CONCLUSION: Our study demonstrated no effect of an educational intervention designed to decrease ER visits. Limitations of our study, however, suggest that further attempts to educate parents in this setting may still be warranted. PMID- 8243910 TI - [Hepatox: database on hepatotoxic drugs]. AB - Hepatox is a data base on the hepatotoxic drugs file published every year in Gastroenterologie Clinique et Biologique. The program was developed under Omnis 7 for Apple computers, and under Visual Basic Professional Toolkit and Code Base for IBM PC and compatibles computers. The data base includes forms of 866 drugs identified by their approved name and those of their 1,300 corresponding proprietary names in France; drugs are distributed among 104 pharmacological classes. It is possible to have instantaneously access to the card of a drug identified by its approved name. Acceding to a drug identified by its proprietary name gives a list of the approved name of its components; going from a name of this list to the correspondent card of hepatoxicity is immediate. It is easy to extract lists of drugs responsible of a type of hepatic injury, and a table of types of hepatic injuries induced by the drugs of a pharmacological class. PMID- 8243911 TI - [Telhepat. Database on hepatotoxic drugs available through Minitel]. PMID- 8243912 TI - [Score of suspected drug-induced acute liver disorders. Presentation of an evaluation sheet]. PMID- 8243913 TI - [Drug-induced acute hepatitis]. PMID- 8243914 TI - [Physiopathology of drug-induced hepatopathies]. PMID- 8243915 TI - [Drug-induced chronic hepatitis and cirrhosis]. PMID- 8243916 TI - [Drug-induced steatosis, phospholipidosis and pseudo-alcoholic liver disorders]. PMID- 8243917 TI - [Drug-induced granulomatous hepatitis]. PMID- 8243918 TI - [Drug-induced vascular lesions of the liver]. PMID- 8243919 TI - [Drug-induced liver tumors]. PMID- 8243920 TI - [Drug-induced biliary tract pathology]. PMID- 8243921 TI - [Occupational liver disorders induced by chemical agents]. PMID- 8243922 TI - [Hepatotoxicity of medicinal plants and plant-based preparations]. PMID- 8243923 TI - [Hepatotoxicity of drugs. An updated bibliographic database of liver disorders and responsible drugs]. PMID- 8243924 TI - [Preliminary evaluation of intrahepatic shunts]. PMID- 8243925 TI - [State of grace for percutaneous portal shunts]. PMID- 8243926 TI - [Transjugular intrahepatic portacaval shunt. Preliminary results]. AB - Transjugular implantation of a metallic prosthesis between main portal branch and a hepatic vein is a new intrahepatic portosystemic shunt method to treat portal hypertension. Forty-nine cirrhotic patients with portal hypertension were treated using this technique. The success rate of implantation was 94%. Mortality due to the procedure was 2%. Early complications decreased with the operator's learning curve consisting of bleeding (20%): digestive hemorrhage (n = 5; 12%), hemobilia (n = 4; 8%), which ceased after withdrawal of post-operative heparin. Early thrombosis following shunt implantation occurred in 13% of the patients. Shunt implantation resulted in a 53 +/- 16% decrease in the portocaval pressure gradient (21 +/- 5 to 10 +/- 4 mmHg). Shunts were performed in 45 patients to prevent a recurrence of variceal bleeding. Follow-up of 41 patients showed 12.2% with variceal rebleeding and 16 patients (39%) with a stenosis requiring a redilation. Ascites improved in 95% of the patients and hepatic encephalopathy occurred in 17%. One year survival was 85%. Controlled studies are needed to determine the indications of this new treatment for ascites and bleeding. PMID- 8243927 TI - [Treatment of hemorrhages by rupture of cardio-tuberous varices with transjugular intrahepatic portasystemic shunt]. AB - Twelve consecutive patients admitted for bleeding from ruptured gastric varices were treated with transjugular intrahepatic portosystemic shunts and followed for a mean of 6 +/- 3 months (range: 8-293 days). The shunt was performed successfully in all 12 patients. The shunt occluded in 3 patients (respectively 19, 101 and 103 days after insertion) of whom one remained asymptomatic and two experienced rebleeding. Four patients presented with acute encephalopathy, spontaneously in two and after rebleeding in two. Three patients died, two after rebleeding and one of septic shock secondary to pneumonia. Overall, 9 patients survived a mean of 211 +/- 92 days with no rebleeding, 8 of whom have not yet experienced any complications. These results suggest that transjugular intrahepatic portosystemic shunts could be useful in treating hemorrhages from ruptured gastric varices and in preventing their recurrence. PMID- 8243928 TI - [Effect of a low-fat diet on the fasting volume and postprandial emptying of the gallbladder]. AB - Little is known of the effects of meal composition on gallbladder emptying and cholecystokinin (CCK) release in man. Gallbladder volumes (measured by means of real time ultrasonography) and plasma CCK levels (determined by radioimmunoassay) were studied in 5 normal subjects, before and after a normal solid-liquid meal, and before and after a low-fat, low-protein, solid-liquid meal after 3 days regimen with low-fat, low-protein meal. Low-fat, low-protein regimen significantly increased gallbladder fasting volume and significantly decreased fasting plasma CCK levels. This suggests that CCK secretion regulates fasting gallbladder volume and that basal CCK secretion depends on diet composition. After a normal meal, gallbladder emptying was biphasic with a 44% volume decrease within the first 15 min followed by slower emptying during the next 60 min with a final volume reaching less than 15% of the fasting volume. After a hypolipidic, hypoproteic meal, the initial 15 min emptying (42%) was the same as after control meals but no further decrease of gallbladder volume was observed. This study shows that the initial phase of post prandial gallbladder contraction is not dependent on meal composition which affects late gallbladder emptying only. We conclude that a low-fat, low-protein diet, increasing gallbladder fasting volume and decreasing gallbladder emptying, may favor gallbladder stasis and therefore increase the risk of gallstone formation. PMID- 8243929 TI - [Tolerance of the liver to normothermic ischemia]. PMID- 8243930 TI - [Is management of the short bowel syndrome specific in the elderly?]. PMID- 8243931 TI - [Posterior fundoplication of 270 degrees. pH results after one and 5 years]. AB - This prospective study compared pre- and postoperative pH monitoring one and five years after a 270 degrees posterior fundoplication. Thirty-six consecutive patients with gastroesophageal reflux were included over a period of 3 years. All patients had preoperative esophagoscopy and 3 hour post-prandial pH monitoring. Thirty-three patients had esophagitis. At one year follow-up, pH monitoring was performed in 31 patients and esophagoscopy in 29. At five years, 23 patients accepted a new pH control. Appraisal of efficacy was assessed by Kaye's acid reflux score (normal < or = 90) and the percentage of time (t) below pH 5 (normal < or = 10%). Among the 23 patients who were submitted 3 times to pH monitoring, Kaye's acid reflux score evolved from 149 (+/- 68) before fundoplication to 1.1 (+/- 2) and 16 (+/- 29) after one and five years respectively. The value of t decreased from 20% (+/- 16) to 0.4% (+/- 0.8) after one year and to 2.5% (+/- 3) after five years. The 270 degrees posterior valve corrects pH recordings at one year. Its efficacy remained steady after 5 years, although there was a slight increase in acid reflux scores. The mean value of these scores was always within normal limits. PMID- 8243932 TI - [Guided puncture-cytology under electronic sectorial ultrasound endoscopy. Results in 26 patients]. AB - The diagnosis of submucosal tumors, extrinsic lesions of gastrointestinal tract and mediastinal or celiac masses can be made by endoscopic ultrasonography (EUS) but histological confirmation (benign or malignant lesion) is still required. The aim of this study was to determine the efficacy of a new method, enabling EUS (curved array transducer)--guided fine needle aspiration of deep lesions. From October 1991 to August 1992, EUS-guided fine needle aspiration was performed in 26 patients in whom a mediastinal mass (6 cases), mediastinal lymph nodes (4 cases), submucosal tumor (5 cases), extrinsic masses (4 cases) and large fold gastric disease with negative endoscopic biopsy (7 cases) was detected by EUS. Diagnostic results were obtained in 19 of 26 patients (73.1%). A malignant tumor was detected in 21 of 26 patients. The EUS-guided fine needle aspiration was positive in 16 cases. The sensitivity for diagnosis of malignancy was 76.2% while specificity was 100%. The development of this new technique in the future needs further study. PMID- 8243933 TI - [Isolated smooth muscle cells of the human colon. Cytophysiological study]. AB - Gross normal specimens of human distal colon were obtained at operation for cancer. Smooth muscle cells were separated from internal and external layers of the muscularis. They were dissociated by digestion with collagenase, isolated and concentrated by successive centrifugations. Colonic smooth muscle cell contraction was measured using various concentrations of carbamylcholine (10(-9) to 10(-4) M); relaxation was tested using atropine (10(-9) to 10(-4) M) on colonic smooth muscle cells pre-contracted by carbamylcholine. Compared with previous descriptions, human smooth muscle cells were smaller than in other species with an enlarged distribution of cell size (30 microns to 150 microns in length). Significant dose-response curves were obtained for both carbamylcholine and atropine. However, 3 original points characterized human colonic smooth muscle cells: a) the cells isolated from the internal layer were significantly more sensitive than those isolated from the external layer (10(-9) M vs 10(-7) M); b) for the muscle cells isolated from both the internal and external layers, small colonic smooth muscle cells were significantly more sensitive. On the other hand, these cells were shown to be located near conjunctive septae, and intramural plexuses; c) analysis of contraction curves demonstrated a more efficient response for colonic smooth muscle cells of the internal layer than for those of the external layer of the muscularis. PMID- 8243934 TI - [Role of serotonin and histamine in the effects of degranulation of mast cells on the colonic motility and the transit. Experimental study in rats]. AB - The aim of this work was to describe the alterations of colonic motility and transit induced by an experimental, histologically verified, degranulation of mast cells, provoked by the compound BrX-537A, and to determine the role of serotonin and histamine by specific antagonists, in the rat. Colonic myoelectrical activity was inhibited by BrX-537A (2 mg/kg IP) in a biphasic manner. The initial profound inhibition, lasting 30 min, during which the frequency of spike bursts decreased from 9.2 +/- 1.1 to 1.4 +/- 0.5/10 min, was followed by a sustained (5 h) period of moderate inhibition (5.2 +/- 0.5 spike bursts/10 min). In the same way, BrX-537A increased the mean retention time of a marker injected in the proximal colon (10.8 +/- 1.4 h vs 7.4 +/- 0.4 h). Neither serotoninergic nor histaminergic antagonists, at a dose of 1 mg/kg IP, modified the primary drastic inhibition of colonic motility during the first 20 minutes. After, a selective time-related blockade of this inhibition was observed. Granisetron blocked the inhibition from the 30th minute on, methysergide from the 120th minute on, and chlorpheniramine, between the 20th and 60th minutes. In conclusion, the inhibitory effect of mast cell degranulation depends on serotonin and histamine release, in a time-related manner, and implicates the H1, 5-HT3 and 5-HT1 or 2 receptors. PMID- 8243935 TI - [Repercussions and undesirable effects of non-steroidal anti-inflammatory agents on the intestine. 1: Experimental data and pathophysiological effects]. PMID- 8243936 TI - [Hepatic calcifications revealing epithelioid hemangioendothelioma]. AB - This report concerns a case of hepatic epithelioid hemangioendothelioma presenting with liver enlargement, multiple hepatic calcifications on the plain abdominal film and bilateral interstitial pneumonia. Epithelioid hemangioendothelioma is an unusual soft tissue tumor of "intermediate malignancy" characterized by a multicentric involvement of the liver and originating from endothelial cells. Histopathologic diagnosis may be difficult because of possible confusion with sclerosing epithelial tumors. PMID- 8243937 TI - [Late portal vein thrombosis caused by hematoma after liver transplantation]. AB - Late portal vein thrombosis of liver grafts is uncommon and is usually revealed by variceal bleeding. We herein report a case of thrombosis discovered in an adult 10 months after liver transplantation, due to stenosis of the portal vein by extrinsic compression, probably secondary to a biopsy-induced hematoma. Development of venous collateral circulation allowed for normal function of the liver graft. PMID- 8243938 TI - [Fatal fulminant hepatitis induced by nilutamide (Anandron)]. AB - The first case of fatal fulminant hepatitis probably caused by nilutamide, a non steroidal antiandrogen derivative, is reported. Nilutamide administration had been continued for 6 days after the occurrence of jaundice, 52 days after beginning of treatment. The rapidly fatal outcome might have been promoted by coadministration of phenobarbital. PMID- 8243939 TI - [Pharyngeal dysphagia and polymyositis]. AB - We report the case of a 70 year-old woman admitted for pharyngeal dysphagia. The diagnosis of polymyositis was made on manometry and histopathological neuromuscular biopsy findings. There were no inflammatory syndrome, muscular enzyme increase or electromyographic abnormalities. The patient was initially treated by prednisone (1 mg/kg/d) with success, but relapsed 12 weeks later and then was put on azathioprine (2 mg/kg/d). Pharyngeal dysphagia can be the only clinical manifestation of polymyositis, usually a systemic disease. PMID- 8243940 TI - [The cost of endoscopy in gastroduodenal ulcer disease]. PMID- 8243941 TI - [Remarks on the cost of endoscopy in gastroduodenal ulcer disease]. PMID- 8243942 TI - [The cost of upper digestive endoscopy: cries and whispers... Bureau du Club de Reflexion des Cabinets de Groupe de Gastroenterologie]. PMID- 8243943 TI - [Cost of esophago-gastro-duodenal endoscopy]. PMID- 8243944 TI - [The cost of endoscopy in gastroduodenal ulcer, or how to rationalize without rationing?]. PMID- 8243945 TI - [Apropos of the cost of endoscopy in gastroduodenal ulcer disease: about the difficulty and the necessity of calculation of costs]. PMID- 8243946 TI - [Hemobilia of late disclosure after placement of transjugular intrahepatic portacaval shunt]. PMID- 8243947 TI - [Hemorrhagic cholecystitis without apparent hemobilia: late complication of hepatic puncture biopsy]. PMID- 8243948 TI - [Section of a Blakemore sound by a patient: 3 cases]. PMID- 8243949 TI - [Encephalopathy associated with hyperammonemia in an alcoholic patient treated with sodium valproate]. PMID- 8243950 TI - [Lichen planus and chronic hepatitis C]. PMID- 8243951 TI - [Acute pseudo-angio-cholitic cholestasis during melphalan therapy]. PMID- 8243952 TI - [Prospective evaluation of two kits for percutaneous endoscopic gastrostomy]. PMID- 8243953 TI - [Value of hyperbaric oxygen in the hemostatic treatment of chronic radiation induced recto-sigmoiditis]. PMID- 8243954 TI - [Are cells of the immune system of the lamina propria of the stomach the targets of anti-ulcer drugs?]. PMID- 8243955 TI - Is adequate good enough? PMID- 8243956 TI - Confidentiality between the dentist and the adolescent. Part I. PMID- 8243957 TI - Infection control in the dental laboratory. PMID- 8243958 TI - Nitrous oxide and fertility. Part II. PMID- 8243959 TI - Analgesic and antibiotic administration during pregnancy. PMID- 8243960 TI - The pregnant dental patient: evaluation and management. PMID- 8243961 TI - Satisfaction level of dental office personnel. PMID- 8243962 TI - Physical evaluation and monitoring devices in dental practice. PMID- 8243963 TI - High blood pressure screening in the dental office: a survey among Dutch dentists. PMID- 8243964 TI - Periodontal and restorative treatment of the class V lesion. PMID- 8243965 TI - [Needle sphincterotomy: a safe method]. AB - We performed needle papillotomy in 26 patients. When the biliary tract was not approach by conventional methods. We used an home made sphincterotomy and begin the incision at the papillar orifice and cut in cephalic direction to the 11-12 o'clock, and stop the cutting when we observe bile flow or choledochus mucosa. The only complication was bleeding in two patients and it stopped with epinephrine injection. There were not perforations or mortality. We conclude that needle papillotomy is a safe and useful method in selected cases. PMID- 8243966 TI - [Measurement of esophageal pH during 24 hours in children. Preliminary report]. AB - A preliminary report is presented regarding the 24 hours continuous pH monitoring of 40 patients from 1 month to 19 years of age with clinical features of gastro esophageal reflux confirmed by at least one of the diagnostic methods utilized (radiology, endoscopy, esophageal biopsy). The equipment used was a MIC GASTROGRAPH with 1.2 and 3 mm catheters according to age and using the methodology suggested by ESPGAN. Patients with reflux index of more than 10% had all other pH indexes and other diagnostic methods abnormal. The area of the curve below pH 4 correlated directly with the presence of esophagitis. Patients with post history of esophageal atresia and caustic esophagitis had marked alterations of pH. PH monitoring with the modern methodology currently available is of great help to quantify severity of acid reflux, to distinguish between physiologic pathologic reflux and to decide treatment. PMID- 8243967 TI - [Urinary epidermal growth factor and serum pepsinogen I in patients with duodenal ulcers]. AB - Absolute and corrected by creatinine excretion urinary Epidermal Growth Factor (EGF) values were determined in 23 duodenal ulcer patients and compared to a control group. Basal serum pepsinogen I levels were measured in the patient group. Absolute urinary EGF values in patients were lower than in control group, such difference however, as such of EGF corrected by urinary creatinine excretion were not statistically significant (p > 0.05). Absolute urinary EGF excretion in male patients was higher than in female patients (p < 0.05), but after establishing the ratio EGF/Creatinine, the difference disappeared. There was no correlation between EGF and Pepsinogen I, it was inverse with age and positive with creatinine excretion. Some mechanisms are considered to explain the urinary EGF normality in these patients. The increasing importance given to EGF in ulcerous diseases and further trends in research are analyzed. PMID- 8243968 TI - [Splenic focal lesion in oncologic patients diagnosed by ultrasound]. AB - Focal lesion of the spleen are infrequent, they appear in 2-4% of patients with malignancies, in our experience we found metastases in nineteen oncologic patients (0.54%) this fact modified the staging prognosis and therapeutic. All the cases were correlated with laparoscopy, fine needle biopsy guided by ultrasound, and or laparotomy. It is emphasized tha the spleen's size was normal in 47.4%. The usual appearance was multiple focal lesions in 89.5% and the sonography pattern of the lesion was variable: Hypoechoic in 63.2% hyperechoic an mixed in 15.8% respectively and cystic in 5.3%. We conclude that ultrasound is an excellent non invasive method to detect focal lesions of the spleen in oncologic patient, where is necessary to know the histology of this lesions, fine needle guided biopsies, can be done safely. PMID- 8243969 TI - [Natural history of viral hepatitis A in Chilean adults: clinical and laboratory aspects]. AB - In many countries, Hepatitis is mainly due to virus. A. When improving life condition in a given population, initially there is a tendency to increase the number of cases in adults. We report clinical and laboratory findings in 87 adults with acute viral Hepatitis A in Chile. The rate man/woman was 1.55/1. Mean age: 23.8 years. Clinical forms: icteric classical (77.01%), cholestatic (10.34%), anicteric (8.05%), biphasic (2.30%) and fulminant (2.30%). From 87 patients in consult 1, 64 were controlled at day 15 (consult 2) and 35 one year later (consult 3). Laboratory (means): ALT (UI/L): 856.8, 111.6 and 20.8 in consult 1, 2 and 3 respectively. Correlation between values of ALT and AST (p < 0.0001). Mean total bilirubin (mg%): 6.6, 2.5 and 0.8 respectively. The evolution of Hepatitis A was favorable with a rapid decrease of clinical signs and normalization of laboratory values within the 3 first weeks of disease. PMID- 8243970 TI - [Measurement of cardia and pylorus distance by endoscopy in children]. AB - We performed upper gastrointestinal endoscopy in 258 children. We measured the distance from the teeth to cardias and pylorus and found, that height correlated with distance from then teeth to cardias. PMID- 8243971 TI - [Macroscopic aspect and depth in early gastric cancer]. AB - Depth of invasion in Early Gastric Cancer (E.G.C.) is very important to be evaluated. We reviewed 78 cases of E.G.C., in patients of our program that were operated on. We established the relationship among three macroscopic variables (localization, size and macroscopic shape) and depth of invasion. We realized that the lesions located in the fundus, though diagnosed less frequently, has more tendency to invade the submucosa (50%), in relation to other localizations. We did not find a significant statistic relation chi 2 = 3.76 P: .58 df:5 between size and invasion. The macroscopic types IIa + IIc and the E.G.C. type III have tendency to further invasion (58.8 and 66.67%). The polypoid lesions Type I and IIa are mostly intramucosal, so they could be treated endoscopically in high risk and old patients. PMID- 8243972 TI - [Leiomyosarcoma of the gastrointestinal tract]. AB - The records and biopsies of 10 patients with the diagnosis of leiomyosarcomas admitted at the Military Hospital "Dr. Carlos Arvelo" from 1962 to 1991, were reviewed. The sample correspond to 0.47% of all gastro-intestinal tumors. Sixty per cent of tumors were localized in the stomach, 20% in the jejunum, 10% in the duodenum and 10% in the colon. Eighty per cent had pain and 60% had bleeding. Sixty per cent of tumors were low grade leiomyosarcomas and 40% were epithelioid leiomyosarcomas. In eight of nine patients operated, resection was done. The survival range was from six months to 28 years with a median of 12 years. The most important prognostic factor was the extension of the tumor, independently of he histologic type. PMID- 8243973 TI - [Physician-patient relations ... an inexhaustible subject]. PMID- 8243974 TI - [Juxtapapillary diverticuli and biliary lithiasis]. AB - Duodenal diverticuli has been associated with biliary stones, specially in the common bile duct. Its presence alters the anatomy of the region of the papilla difficulting its identification and cannulation. ERCP was performed in 800 patients with suspicion of bilio-pancreatic disease, 60 (7.5%) of these patients (39 male and 21 female) had juxtapapillary diverticuli (single diverticuli in 52, 2 in 7 and 3 in 1 patient). In these patient cannulation of the common bile duct was possible in 43 (72%), only the pancreatic duct in 5 (8%) and no cannulation in 12 (20%). Common bile duct stones were found in 22 patients, papillotomy was indicated in 14 patients and was performed in 12, in 2 patients it was not possible to position the papillotomy. PMID- 8243975 TI - Malaria chemotherapy: resistance to quinoline containing drugs in Plasmodium falciparum. AB - Resistance to quinoline containing drugs, particularly chloroquine (CQ), is a major impediment to the successful chemotherapy and prophylaxis of malaria. CQ resistant parasites fail to accumulate as much drug as their sensitive counterparts and two major hypotheses have been proposed to account for this phenomenon. CQ-resistant parasites are thought to maintain lower intracellular drug levels by means of an active efflux system, similar to that found in multi drug resistant cancer cells, despite major differences in both the genetic and biochemical manifestations of drug resistance in the two cell types. Alternatively, CQ-resistance could be linked to a defective CQ uptake mechanism, possibly an impaired acidification process in the food vacuole of the resistant parasite. These two theories are discussed in detail in the following review. The potential of pharmacological intervention to override these resistance mechanisms is also discussed. PMID- 8243976 TI - Identification of a nuclear pheromone-sensitive protein kinase not identical to p34CDC28 in Saccharomyces cerevisiae. AB - Nuclei of Saccharomyces cerevisiae cells contain a protein kinase, the activity of which is drastically reduced in response to an activation of the mating signal pathway by pheromone. Inhibition of this pheromone-sensitive kinase is also observed under conditions of constitutive activation of the signal pathway in a temperature-sensitive cdc70 mutant. The enzyme, which by SDS-PAGE has a molecular mass of 34,500 Da, is a protein serine kinase that phosphorylates several endogenous substrates in nuclear extracts. The activity of this kinase is temperature-resistant in a temperature-sensitive cdc28 mutant, indicating that it is not identical to p34CDC28, the catalytic component of the cell cycle protein kinase complex. PMID- 8243977 TI - Arabinan degrading enzymes from Aspergillus nidulans: induction and purification. AB - The presence in Aspergillus nidulans of two enzymes related to the Aspergillus niger endo-arabinase and alpha-L-arabinofuranosidase B has been established using antibodies against the purified A. niger enzymes. Moreover, the absence of an equivalent in A. nidulans to the alpha-L-arabinofuranosidase A of A. niger has been confirmed both at the protein and at the DNA level. Both A. nidulans arabinases have been purified and physico-chemically and kinetically characterized. They have a much higher temperature optimum than the corresponding A. niger enzymes. The pattern of induction has been studied on media containing different carbon sources showing an important role of L-arabitol in the induction of these enzymes. PMID- 8243978 TI - Clostridium bifermentans serovar malaysia: characterization of putative mosquito larvicidal proteins. AB - The toxicity of Clostridium bifermentans serovar malaysia to mosquito larvae is due to protein toxins, belonging to a novel class of insecticidal toxins. Toxic extracts contains three major proteins of 66, 18 and 16 kDa. The 18-kDa and 16 kDa proteins are probably involved in toxicity. They are synthesised during sporulation, concomitant with activity. They are absent from non-toxic strains of C. bifermentans and are present at very low levels in non-toxic C. bifermentans serovar malaysia cultures produced at 42 degrees C. The 66-kDa protein is present throughout the growth phases of C. bifermentans serovar malaysia, and an immunologically related 66-kDa protein is present in non-toxic C. bifermentans strains. PMID- 8243979 TI - Synthesis of the Rhodopseudomonas viridis holo-cytochrome c2 in Paracoccus denitrificans. AB - The gene encoding the Rhodopseudomonas viridis cytochrome c2 (cycA) has been introduced on a broad host range vector into Paracoccus denitrificans, leading to high-level expression of the holo-cytochrome with the heme moiety covalently attached to the apoprotein. The cytochrome was demonstrated to reside in the periplasmic space of the host cell. In contrast to R. viridis, aerobic rather than anaerobic growth conditions led to higher production levels of the holo cytochrome in P. denitrificans. This heterologous expression system provides a suitable genetic background for the functional expression and mutagenesis of polypeptides involved in bacterial photosynthesis, offering the possibility of detailed structural and functional investigation. PMID- 8243980 TI - A method for enucleation of Saccharomyces cerevisiae. AB - The first method for enucleation of yeast Saccharomyces cerevisiae is reported. Various strains, including some killer strain and respiratory-deficient mutants of Saccharomyces cerevisiae were enucleated after treatment with cytochalasin B. Removal of nuclei from protruding sphaeroplasts was induced by centrifugation in a Percoll density gradient. The enucleation yield (which averaged about 80%) and the quality of the cytoplasts were best when the yeast culture had been synchronized with nocodazole before the preparation. The presence of 1 mM CaCl2 and ATP (10 microM) in the enucleation medium prevented the formation of fragile products or aggregation. Cytoplasts could be stored for at least 1 day without visible deterioration. PMID- 8243981 TI - Use of RAPD-PCR to isolate a species specific DNA probe for Phytophthora cinnamomi. AB - The products of RAPD-PCR amplification of Phytophthora cinnamomi DNA were separated by electrophoresis in agarose. Parallel Southern blots of the gels were hybridized with nick translated DNA from different species of Phytophthora. Fragments that hybridized specifically to P. cinnamomi DNA were identified. These fragments were purified and cloned into pUC18. Their specificity for P. cinnamomi was confirmed. PMID- 8243982 TI - Primary structure analysis of a duplicated region in the amplifiable AUD6 locus of Streptomyces ambofaciens DSM40697. AB - In Streptomyces ambofaciens, an amplifiable unit of DNA (AUD6) contains two homologous sequences, one located on the right extremity of the AUD (S1R), the other being internal (IHS). This paper presents the molecular analysis of this duplication. The nucleotide sequences are almost identical (95%) and each contains an ORF of about 330 codons, the two ORFs being nearly identical. The two hypothetical proteins, deduced from these sequences, show about 30% identity with different bacterial repressors. They also show a particularly strong similarity (90% identity between the full-length sequences) with hypothetical proteins of Streptomyces lividans 66 encoded by sequences also present on an amplifiable DNA region (AUD1). PMID- 8243983 TI - Kinetic properties of yeast lysine permeases coded by genes on multi-copy vectors. AB - Amplification of the LYP1 transport system in the plasma membrane of Saccharomyces cerevisiae did not change the substrate specificity, the affinity and the pH optimum of the transport system for lysine, but significantly increased the uptake velocity and accumulation ratio. The dependence of active lysine uptake and accumulation on pH is probably given by the properties of the permease itself rather than by the value of available protonmotive force. PMID- 8243984 TI - Chemotaxis away from thiocyanic and isothiocyanic esters in Pseudomonas aeruginosa. AB - Negative chemotaxis, the movement of organisms away from chemicals, was investigated in Pseudomonas aeruginosa using a rapid videotape method. Digital image processing was used to detect changes in bacterial numbers near the mouth of a capillary containing a test chemical. P. aeruginosa was found to be repelled by thiocyanic and isothiocyanic esters including allyl isothiocyanate, ethyl thiocyanate, methyl isothiocyanate and methyl thiocyanate. Particularly, the movement of bacteria away from methyl thiocyanate was so drastic that bacterial numbers near the mouth of the capillary decreased by approximately 80% within 30 s after the start of observation. Mutant strains, fully motile but lacking positive chemotaxis, did not escape away from the esters, suggesting a common mechanism between positive and negative chemotaxes in this organism. PMID- 8243985 TI - Cell-associated hemagglutinin of classical vibrio cholerae O1 with reference to intestinal adhesion. AB - Vibrio cholerae O1 86B3 biovar cholerae has at least two types of cell-associated hemagglutinin. One is cell wall-associated and L-fucose sensitive, whereas the other is pili-associated and D-mannose sensitive. A pilus rich variant of 86B3 and a poorly piliated parent strain adhered equally to the rabbit intestine. This adhesion was inhibited by L-fucose but not by D-mannose. A Fab fraction prepared from anti-pilus antibody did not inhibit the adhesion. These results suggest that not the pili but a colonization factor located in the outer membrane of the organisms plays a role in intestinal adhesion. PMID- 8243986 TI - Extracellular glycoprotein specific for Saccharomyces sensu stricto. AB - Using affinity-purified rabbit polyclonal antibodies against an extracellular mannoprotein (gp400) of Saccharomyces cerevisiae, the presence of immunohomologic proteins with similar electrophoretic mobility was shown in the culture medium of S. bayanus, S. paradoxus and S. pastorianus. Cross-reactive bands with different electrophoretic behaviour were observed for S. dairensis, S. exiguus, S. kluyveri, S. unisporus and also for the species moved from Saccharomyces to Arxiozyma, Kluyveromyces, Pachytichospora, Torulaspora and Zygosaccharomyces, in contrast to ascosporous yeasts of other genera in which these proteins were not found. PMID- 8243987 TI - Phylogenetic analysis of Desulfotomaculum thermobenzoicum using polymerase chain reaction-amplified 16S rRNA-specific DNA. AB - The 16S rRNA gene of the thermophilic sulfate-reducing bacterium Desulfotomaculum thermobenzoicum was amplified by polymerase chain reaction using two eubacterial consensus oligodeoxynucleotide primers flanking the majority of the 16S rRNA gene, cloned, and sequenced. Phylogenetic analysis revealed that D. thermobenzoicum belongs to the Gram-positive (low G + C content) branch and is more related to the thermophilic sulfate-reducing bacterium, D. australicum than the moderate thermophile D. nigrificans, or the mesophiles D. orientis, and D. ruminis. This relationship is further strengthened by the presence of an unusual idiosyncrasy in helix 6 of the 16S rRNA gene of D. thermobenzoicum resembling that of D. australicum but not found in other desulfotomacula species and in any other bacteria sequenced to date. PMID- 8243988 TI - Towards a phylogeny of the clostridia based on 16S rRNA sequences. AB - The 16S rRNA sequences of 17 species of the genus Clostridium were determined by direct sequencing of their PCR amplified genes. The sequences were aligned with those from other known clostridial species and representative low G + C Gram positive relatives, and a phylogenetic tree was constructed. It was evident from the comparative sequence analysis that the genus Clostridium as presently constituted is phylogenetically extremely heterogeneous. This study corroborates and extends earlier findings in showing that many non-sporeforming bacteria are phylogenetically closely intermixed with Clostridium species. The taxonomic implications of the phylogenetic findings are discussed. PMID- 8243989 TI - Impairment of nucleoid segregation and cell division at high osmolarity in a strain of Escherichia coli overproducing the chaperone DnaK. AB - Escherichia coli strain WM1390, which overproduces 20-fold the chaperone protein DnaK, was able to grow exponentially without apparent abnormality in 300 mosM medium. In contrast, it showed both aberrant nucleoid segregation and strong inhibition of septation when shifted to high osmolarity. These impairments could not be accounted for by a bacteriocidal effect of the high DnaK content. Rather, the DnaK content appeared to promote faster growth than that of the parent C600, at least at high osmolarity in the presence of the osmoprotectant glycine betaine. PMID- 8243990 TI - Psychosocial risk factors for HIV transmission in male drug abusers. AB - We studied the psychosocial risk factors associated with Black, Hispanic, and White (N = 257) male intravenous drug abusers' needle-sharing behavior with familiar individuals and with strangers. Approximately half the subjects were HIV+. The findings supported a mediational model in that risk factors in the family domain were associated with peer and personality risk factors conducive to needle sharing that, in turn, were related to needle-sharing behavior. PMID- 8243991 TI - Roland Thaxter's legacy and the origins of multicellular development. PMID- 8243992 TI - Screens for extragenic mutations that fail to complement act1 alleles identify genes that are important for actin function in Saccharomyces cerevisiae. AB - Null mutations in SAC6 and ABP1, genes that encode actin-binding proteins, failed to complement the temperature-sensitive phenotype caused by a mutation in the ACT1 gene. To identify novel genes whose protein products interact with actin, mutations that fail to complement act1-1 or act1-4, two temperature-sensitive alleles of ACT1, were isolated. A total of 14 extragenic noncomplementing mutations and 12 new alleles of ACT1 were identified in two independent screens. The 14 extragenic noncomplementing mutations represent alleles of at least four different genes, ANC1, ANC2, ANC3 and ANC4 (Actin NonComplementing). Mutations in the ANC1 gene were shown to cause osmosensitivity and defects in actin organization; phenotypes that are similar to those caused by act1 mutations. We conclude that the ANC1 gene product plays an important role in actin cytoskeletal function. The 12 new alleles of ACT1 will be useful for further elucidation of the functions of actin in yeast. PMID- 8243993 TI - Genetic evidence for functional interactions between actin noncomplementing (Anc) gene products and actin cytoskeletal proteins in Saccharomyces cerevisiae. AB - We describe here genetic interactions between mutant alleles of Actin NonComplementing (ANC) genes and actin (ACT1) or actin-binding protein (SAC6, ABP1, TPM1) genes. The anc mutations were found to exhibit allele-specific noncomplementing interactions with different act1 mutations. In addition, mutant alleles of four ANC genes (ANC1, ANC2, ANC3 and ANC4) were tested for interactions with null alleles of actin-binding protein genes. An anc1 mutant allele failed to complement null alleles of the SAC6 and TPM1 genes that encode yeast fimbrin and tropomyosin, respectively. Also, synthetic lethality between anc3 and sac6 mutations, and between anc4 and tpm1 mutations was observed. Taken together, the above results strongly suggest that the ANC gene products contribute to diverse aspects of actin function. Finally, we report the results of tests of two models previously proposed to explain extragenic noncomplementation. PMID- 8243994 TI - Point mutations that separate the role of Saccharomyces cerevisiae centromere binding factor 1 in chromosome segregation from its role in transcriptional activation. AB - Centromere binding factor 1 (Cbf1p or CP1) binds to the CDEI region of Saccharomyces cerevisiae centromeres and is a member of the basic helix-loop helix (bHLH) class of proteins. Deletion of the gene encoding Cbf1p results in an increased frequency of chromosome loss, hypersensitivity to low levels of microtubule disrupting drugs (such as thiabendazole and benomyl) and methionine auxotrophy. By polymerase chain reaction-based random mutagenesis of the CBF1 gene we have obtained a number of mutant alleles that make full-length protein with impaired function. The mutations in these alleles are clustered in or just downstream from the bHLH domain. Among the alleles obtained was a class that was more compromised for transcriptional activation and a class that was more compromised for chromosome loss and thiabendazole hypersensitivity. These results indicate that at least some aspects of the role of Cbf1p in chromosome segregation and transcriptional activation are distinct. In contrast, increased chromosome loss and thiabendazole hypersensitivity were not separated in any of the alleles, suggesting that these phenotypes reflect the same mechanistic defect. These observations are consistent with a model that suggests that one role of Cbf1p in chromosome segregation may be to improve the efficiency with which contact between the kinetochore and spindle microtubules is established or maintained. PMID- 8243995 TI - Mixed segregation and recombination of chromosomes and YACs during single division meiosis in spo13 strains of Saccharomyces cerevisiae. AB - Diploid yeast strains, homozygous for the mutation spo13, undergo a single division meiosis and form dyads (two spores held together in one ascus). Dyad analysis of spo13/spo13 strains with centromere-linked markers on five different chromosomes and on a pair of human DNA YACs shows that: (a) in spo13 meiosis, chromosomes undergo mixed segregation, namely some chromosomes segregate reductionally whereas others, in the same cell, segregate equationally; (b) different chromosomes exhibit different segregation tendencies; (c) recombination between homologous chromosomes might not determine that a bivalent undergoes reductional rather than equational segregation. PMID- 8243996 TI - A rare tRNA-Arg(CCU) that regulates Ty1 element ribosomal frameshifting is essential for Ty1 retrotransposition in Saccharomyces cerevisiae. AB - Translation of the yeast retrotransposon Ty1 TYA1(gag)-TYB1(pol) gene occurs by a +1 ribosomal frameshifting event at the sequence CUU AGG C. Because overexpression of a low abundance tRNA-Arg(CCU) encoded by the HSX1 gene resulted in a reduction in Ty1 frameshifting, it was suggested that a translational pause at the AGG-Arg codon is required for optimum frameshifting. The present work shows that the absence of tRNA-Arg(CCU) affects Ty1 transposition, translational frameshifting, and accumulation of mature TYB1 proteins. Transposition of genetically tagged Ty1 elements decreases at least 50-fold and translational frameshifting increases 3-17-fold in cells lacking tRNA-Arg(CCU). Accumulation of Ty1-integrase and Ty1-reverse transcriptase/ribonuclease H is defective in an hsx1 mutant. The defect in Ty1 transposition is complemented by the wild-type HSX1 gene or a mutant tRNA-Arg(UCU) gene containing a C for T substitution in the first position of the anticodon. Overexpression of TYA1 stimulates Ty1 transposition 50-fold above wild-type levels when the level of transposition is compared in isogenic hsx1 and HSX1 strains. Thus, the HSX1 gene determines the ratio of the TYA1 to TYA1-TYB1 precursors required for protein processing or stability, and keeps expression of TYB1 a rate-limiting step in the retrotransposition cycle. PMID- 8243997 TI - Responsiveness to exogenous cAMP of a Saccharomyces cerevisiae strain conferred by naturally occurring alleles of PDE1 and PDE2. AB - The Saccharomyces cerevisiae strain P-28-24C, from which cAMP requiring mutants derived, responded to exogenously added cAMP. Upon the addition of cAMP, this strain showed phenotypes shared by mutants with elevated activity of the cAMP pathway. Genetic analysis involving serial crosses of this strain to a strain with another genetic background revealed that the responsiveness to cAMP results from naturally occurring loss-of-function alleles of PDE1 and PDE2, which encode low and high affinity cAMP phosphodiesterases, respectively. In addition, P-28 24C was found to carry a mutation conferring slow growth that lies in CYR1, which encodes adenylate cyclase, and the slow growth phenotype caused by the cyr1 mutation was suppressed by the pde2 mutation. Therefore P-28-24C is fortuitously a pde1 pde2 cyr1 triple mutant. Responsiveness to cAMP conferred by pde mutations suggests that S. cerevisiae cells are permeable to cAMP to some extent and that the apparent absence of effect of exogenously added cAMP on wild-type cells is due to immediate degradation by cAMP phosphodiesterases. PMID- 8243998 TI - Identification and cloning of the CHL4 gene controlling chromosome segregation in yeast. AB - A collection of chl mutants characterized by decreased fidelity of chromosome transmission and by minichromosome nondisjunction in mitosis was examined for the ability to maintain nonessential dicentric plasmids. In one of the seven mutants analyzed, chl4, dicentric plasmids did not depress cell division. Moreover, nonessential dicentric plasmids were maintained stably without any rearrangements during many generations in the chl4 mutant. The rate of mitotic heteroallelic recombination in the chl4 mutant was not increased compared to that in an isogenic wild-type strain. Analysis of the segregation of a marked chromosome indicated that sister chromatid nondisjunction and sister chromatid loss contributed equally to chromosome malsegregation in the chl4 mutant. A genomic clone of CHL4 was isolated by complementation of the chl4-1 mutation and was physically mapped to the right arm of chromosome IV near the SUP2 gene. Nucleotide sequence analysis of CHL4 clone revealed a 1.4-kb open reading frame coding for a 53-kD predicted protein which does not have homology to published proteins. A strain containing a null allele of CHL4 is viable under standard growth conditions but has a temperature-sensitive phenotype (conditional lethality at 36 degrees). We suggest that the CHL4 gene is required for kinetochore function in the yeast Saccharomyces cerevisiae. PMID- 8243999 TI - The Drosophila melanogaster suppressor of Hairy-wing zinc finger protein has minimal effects on gene expression in Saccharomyces cerevisiae. AB - Many mutations in Drosophila melanogaster are gypsy retrotransposon insertions. Gypsy binds the protein (SUHW) encoded by the suppressor of Hairy-wing [su(Hw)] gene, and SUHW alters expression of surrounding genes. When gypsy is between an enhancer and promoter, SUHW blocks activation of transcription by the enhancer. Additionally, when gypsy is downstream of a promoter in a parallel orientation, SUHW increases truncation of transcripts at the poly(A) site in the gypsy 5' long terminal repeat, thereby decreasing the gene transcript levels. The effects of SUHW appear to involve fundamental and general mechanisms controlling gene expression because SUHW potentiates other poly(A) sites and blocks several enhancers in Drosophila. To investigate these mechanisms, SUHW was expressed in Saccharomyces cerevisiae. Although SUHW enters the nucleus and binds DNA in yeast, it has surprisingly minor effects on utilization of the CYC1 poly(A) site and transcription activation by a GAL upstream activation sequence. These observations indicate that the observed effects of SUHW on gene expression in Drosophila require specific interactions with other factors that are absent or unrecognizable in yeast. PMID- 8244000 TI - De novo methylation of repeated sequences in Coprinus cinereus. AB - We have examined the stability of duplicated DNA sequences in the sexual phase of the life cycle of the basidiomycete fungus, Coprinus cinereus. We observed premeiotic de novo methylation in haploid nuclei containing either a triplication, a tandem duplication, or an ectopic duplication. Methylation changes were not observed in unique sequences. Repeated sequences underwent methylation changes during the dikaryotic stage. In one cross, 27% of the segregants exhibited methylation-directed gene inactivation. However, all auxotrophs eventually reverted to prototrophy. C to T transition mutation were not observed in this study. Our studies also revealed one inversion that occurred in 50% of the segregants in a single triplication cross, and a single pop-out event that occurred during vegetative growth. These alterations were similar to changes reported in experiments with duplicated sequences in Neurospora crassa and Ascobolus immersus. However, significant differences were also noted. First, the extent of methylation was much less in C. cinereus than in the other two fungi. Second, CpG sequences appeared to be the preferred targets of methylation. PMID- 8244001 TI - Population structure in Daphnia obtusa: quantitative genetic and allozymic variation. AB - Quantitative genetic analyses for body size and for life history characters within and among populations of Daphnia obtusa reveal substantial genetic variance at both hierarchical levels for all traits measured. Simultaneous allozymic analysis on the same population samples indicate a moderate degree of differentiation: GST = 0.28. No associations between electrophoretic genotype and phenotypic characters were found, providing support for the null hypothesis that the allozymic variants are effectively neutral. Therefore, GST can be used as the null hypothesis that neutral phenotypic evolution within populations led to the observed differentiation for the quantitative traits, which I call QST. The results of this study provide evidence that natural selection has promoted diversification for body size among populations, and has impeded diversification for relative fitness. Analyses of population differentiation for clutch size, age at reproduction, and growth rate indicate that neutral phenotypic evolution cannot be excluded as the cause. PMID- 8244002 TI - Cloning of flagellar genes in Chlamydomonas reinhardtii by DNA insertional mutagenesis. AB - Chlamydomonas is a popular genetic model system for studying many cellular processes. In this report, we describe a new approach to isolate Chlamydomonas genes using the cloned nitrate reductase gene (NIT1) as an insertional mutagen. A linearized plasmid containing the NIT1 gene was introduced into nit1 mutant cells by glass-bead transformation. Of 3000 Nit+ transformants examined, 74 showed motility defects of a wide range of phenotypes, suggesting that DNA transformation is an effective method for mutagenizing cells. For 13 of 15 such motility mutants backcrossed to nit- mutant strains, the motility phenotype cosegregated with the Nit+ phenotype, indicating that the motility defects of these 13 mutants may be caused by integration of the plasmid. Further genetic analysis indicated that three of these mutants contained alleles of previously identified loci: mbo2 (move backward only), pf13 (paralyzed flagella) and vfl1 (variable flagellar number). Three other abnormal-flagellar-number mutants did not map to any previously described loci at which mutations produce similar phenotypes. Genomic sequences flanking the integrated plasmid in the mbo2 and vfl1 mutants were isolated and used as probes to obtain wild-type genomic clones, which complemented the motility defects upon transformation into cells. Our results demonstrate the potential of this new approach for cloning genes identified by mutation in Chlamydomonas. PMID- 8244003 TI - Sequence requirements for myosin gene expression and regulation in Caenorhabditis elegans. AB - Four Caenorhabditis elegans genes encode muscle-type specific myosin heavy chain isoforms: myo-1 and myo-2 are expressed in the pharyngeal muscles; unc-54 and myo 3 are expressed in body wall muscles. We have used transformation-rescue and lacZ fusion assays to determine sequence requirements for regulated myosin gene expression during development. Multiple tissue-specific activation elements are present for all four genes. For each of the four genes, sequences upstream of the coding region are tissue-specific promoters, as shown by their ability to drive expression of a reporter gene (lacZ) in the appropriate muscle type. Each gene contains at least one additional tissue-specific regulatory element, as defined by the ability to enhance expression of a heterologous promoter in the appropriate muscle type. In rescue experiments with unc-54, two further requirements apparently independent of tissue specificity were found: sequences within the 3' non-coding region are essential for activity while an intron near the 5' end augments expression levels. The general intron stimulation is apparently independent of intron sequence, indicating a mechanistic effect of splicing. To further characterize the myosin gene promoters and to examine the types of enhancer sequences in the genome, we have initiated a screen of C. elegans genomic DNA for fragments capable of enhancing the myo-2 promoter. The properties of enhancers recovered from this screen suggest that the promoter is limited to muscle cells in its ability to respond to enhancers. PMID- 8244004 TI - Genetic characterization of tube and pelle, genes required for signaling between Toll and dorsal in the specification of the dorsal-ventral pattern of the Drosophila embryo. AB - tube and pelle are two of the maternally transcribed genes required for dorsal ventral patterning of the Drosophila embryo. Females homozygous for strong alleles of tube or pelle produce embryos that lack all ventral and lateral embryonic pattern elements. By analyzing the phenotypes caused by 24 pelle and 9 tube alleles, we have defined characteristic features of the two genes, including the extremely variable phenotypes of a number of tube alleles and the antimorphic character of a number of pelle alleles. Double mutant females carrying dominant ventralizing alleles of Toll and dorsalizing alleles of tube or pelle produce dorsalized embryos, suggesting that tube and pelle act downstream of the membrane protein Toll in the signaling pathway that defines the embryonic dorsal-ventral pattern. Both tube and pelle are also important zygotically for survival: at least 30% of the zygotes lacking either tube or pelle die before adult stages, while 90-95% of tube- pelle- double mutant zygotes die. We discuss the phenotypes of tube-pelle double mutants in the context of whether the two proteins interact directly. PMID- 8244005 TI - Genetic and molecular analysis of the X chromosomal region 14B17-14C4 in Drosophila melanogaster: loss of function in NONA, a nuclear protein common to many cell types, results in specific physiological and behavioral defects. AB - We have performed a genetic analysis of the 14C region of the X chromosome of Drosophila melanogaster to isolate loss of function alleles of no-on-transient A (nonA; 14C1-2; 1-52.3). NONA is a nuclear protein common to many cell types, which is present in many puffs on polytene chromosomes. Sequence data suggest that the protein contains a pair of RNA binding motifs (RRM) found in many single strand nucleic acid binding proteins. Hypomorphic alleles of this gene, which lead to aberrant visual and courtship song behavior, still contain normally distributed nonA RNA and NONA protein in embryos, and in all available alleles NONA protein is present in puffs of third instar larval polytene chromosomes. We find that complete loss of this general nuclear protein is semilethal in hemizygous males and homozygous cell lethal in the female germline. Surviving males show more extreme defects in nervous system function than have been described for the hypomorphic alleles. Five other essential genes that reside within this region have been partially characterized. PMID- 8244006 TI - The effects of interspecific Y chromosome replacements on hybrid sterility within the Drosophila simulans clade. AB - We attempted to introgress Y chromosomes between three sibling species of Drosophila: D. simulans, D. sechellia and D. mauritiana. Four D. sechellia Y chromosomes were introgressed into D. simulans without loss of fertility whereas the four reciprocal introgressions (D. simulans Y introgressed into D sechellia) all result in sterility. Both reciprocal Y introgressions of D. simulans and D. mauritiana (four of each) also result in sterility. Compared with D. simulans males, the males with the D. sechellia Y chromosome in D. simulans background had lower productivity but only after multiple matings with virgin females. These males also were inferior compared with pure species males in sperm displacement and/or remating ability. The two different Y genotype males, however, were comparable in viability, longevity and mating success in female choice tests. We also use our results to estimate the effective number of autosomal loci interacting with X-linked genes to produce hybrid male sterility. PMID- 8244007 TI - Genetic dissection of pointed, a Drosophila gene encoding two ETS-related proteins. AB - The Drosophila gene pointed (pnt) is required for the differentiation of a number of tissues during embryogenesis, including the ventral ectoderm, the nervous system, the tracheal system and certain muscle fibers. The phenotypes associated with strong pointed alleles are reflected by a complex pointed expression pattern during embryogenesis. Two promoters, P1 and P2, separated by some 50 kb of genomic sequences, direct the transcription of two different transcript forms, encoding two different proteins related to the ETS family of transcription factors. To assess the individual functions of the two different pointed protein forms, we have generated new pointed alleles affecting either the P1 or the P2 transcript, termed P1 and P2 alleles, respectively. Genetic analysis reveals partial heteroallelic complementation between certain pointed P1 and P2 alleles. Surviving trans-heterozygous flies have rough eyes, abnormal wings and halters, suggesting a requirement for pointed function during their imaginal disc development. Further genetic analysis demonstrates that expression of a given pointed P2 allele depends on trans-acting transcriptional regulatory sequences. We have identified two chromosomal domains with opposite regulatory effects on the transcriptional activity of the pointed P2 promoter, one trans-activates and the other trans-represses pointed P2 expression. By deletion mapping we were able to localize these control regions within the 5' region of the pointed P2 transcript. PMID- 8244008 TI - Deletions and amplifications of tandemly arranged ribosomal 5S genes internal to a P element occur at a high rate in a dysgenic context. AB - We observed unusual kinds of rearrangements within tandemly clustered 5S genes internal to a P element in dysgenic context. Rearranged P transposons, initially containing eight 5S genes, were found to display discrete numbers of 5S genes, from 4 up to 17 units. Precise deletions and amplifications occurred at a high rate (40%), at both original and new insertion sites. These events can be explained by a "cut and paste" transposition model. Possible links between rearrangements due to dysgenic-like processes and concerted evolution are discussed. PMID- 8244009 TI - Population genetics of tandem repeats in centromeric heterochromatin: unequal crossing over and chromosomal divergence at the Responder locus of Drosophila melanogaster. AB - The Responder (Rsp) locus in Drosophila melanogaster is the target locus of segregation distortion and is known to be comprised of a tandem array of 120-bp repetitive sequences. In this study, we first determined the large scale molecular structure of the Rsp locus, which extends over a region of 600 kb on the standard sensitive (cn bw) chromosome. Within the region, small Rsp repeat arrays are interspersed with non-Rsp sequences and account for 10-20% of the total sequences. We isolated and sequenced 32 Rsp clones from three different chromosomes. The main results are: (1) Rsp repeats isolated from the same chromosome are not more similar than those from different chromosomes. This implies either that there are more homologous exchanges at the Rsp locus than expected or, alternatively, that the second chromosomes of D. melanogaster have diverged from one another more recently at the centromeric heterochromatin than at the nearby euchromatin. (2) The repeats usually have a dimeric structure with an average difference of 16% between the left and right halves. The differences allow us to easily identify the products of unequal exchanges. Despite the large differences between the two halves, exchanges have occurred frequently and the majority of them fall within a 29-bp interval of identity between the two halves. Our data thus support the suggestion that recombination depends on short stretches of complete identity rather than long stretches of general homology. (3) Frequent unequal crossover events obscure the phylogenetic relationships between repeats; therefore, different parts of any single repeat could often have different phylogenetic histories. The high rate of unequal crossing over may also help explain the evolutionary dynamics of the Rsp locus. PMID- 8244010 TI - Toward a molecular genetic analysis of spermatogenesis in Drosophila melanogaster: characterization of male-sterile mutants generated by single P element mutagenesis. AB - We describe 83 recessive autosomal male-sterile mutations, generated by single P element mutagenesis in Drosophila melanogaster. Each mutation has been localized to a lettered subdivision of the polytene map. Reversion analyses, as well as complementation tests using available chromosomal deficiencies, indicate that the insertions are responsible for the mutant phenotypes. These mutations represent 63 complementation groups, 58 of which are required for spermatogenesis. Phenotypes of the spermatogenesis mutants were analyzed by light microscopy. Mutations in 12 loci affect germline proliferation, spermatocyte growth, or meiosis. Mutations in 46 other loci disrupt differentiation and maturation of spermatids into motile sperm. This collection of male-sterile mutants provides the basis for a molecular genetic analysis of spermatogenesis. PMID- 8244011 TI - Effects of transposable elements on the expression of the forked gene of Drosophila melanogaster. AB - The products of the forked gene are involved in the formation and/or maintenance of a temporary fibrillar structure within the developing bristle rudiment of Drosophila melanogaster. Mutations in the forked locus alter this structure and result in aberrant development of macrochaetae, microchaetae and trichomes. The locus has been characterized at the molecular level by walking, mutant characterization and transcript analysis. Expression of the six forked transcripts is temporally restricted to mid-late pupal development. At this time, RNAs of 6.4, 5.6, 5.4, 2.5, 1.9 and 1.1 kilobases (kb) are detected by Northern analysis. The coding region of these RNAs has been found to be within a 21-kb stretch of genomic DNA. The amino terminus of the proteins encoded by the 5.4- and 5.6-kb forked transcripts contain tandem copies of ankyrin-like repeats that may play an important role in the function of forked-encoded products. The profile of forked RNA expression is altered in seven spontaneous mutations characterized during this study. Three forked mutations induced by the insertion of the gypsy retrotransposon contain a copy of this element inserted into an intron of the gene. In these mutants, the 5.6-, 5.4- and 2.5-kb forked mRNAs are truncated via recognition of the polyadenylation site in the 5' long terminal repeat of the gypsy retrotransposon. These results help explain the role of the forked gene in fly development and further our understanding of the role of transposable elements in mutagenesis. PMID- 8244012 TI - The pleiotropic function of Delta during postembryonic development of Drosophila melanogaster. AB - Analysis of the development of Delta (Dl) temperature-sensitive mutants pulsed at restrictive temperature during larval and pupal stages reveals multiple phenocritical periods during which reduction of Dl function affects viability and development of adult structures. Dl function is required during the third larval instar for post-pupal viability and during the first day of pupal development for viability through eclosion. Dl function is required biphasically for the development of sensory bristles. Earlier pulses lead to bristle multiplication and later pulses lead to bristle loss. The exact intervals during which multiplication and loss are induced vary for different bristles. Dl function is also required for development of most, if not all, cell types in the retina. Different pulses result in reduction in eye size, scarring, and glossiness, as well as multiplication and loss of interommatidial bristles. We also define intervals during which Dl function is required for aspects of leg and wing development. Phenocritical periods for Dl function are temporally coincident with those previously reported for Notch (N), consistent with the hypothesis that the proteins encoded by Dl and N interact throughout development to assure correct specification of cell fates in a variety of imaginal tissues. PMID- 8244013 TI - Estimates of linkage disequilibrium and the recombination parameter determined from segregating nucleotide sites in the alcohol dehydrogenase region of Drosophila pseudoobscura. AB - The alcohol dehydrogenase (Adh) region of Drosophila pseudoobscura, which includes the two genes Adh and Adh-Dup, was used to examine the pattern and organization of linkage disequilibrium among pairs of segregating nucleotide sites. A collection of 99 strains from the geographic range of D. pseudoobscura were nucleotide-sequenced with polymerase chain reaction-mediated techniques. All pairs of the 359 polymorphic sites in the 3.5-kb Adh region were tested for significant linkage disequilibrium with Fisher's exact test. Of the 74,278 pairwise comparisons of segregating sites, 127 were in significant linkage disequilibrium at the 5% level. The distribution of five linkage disequilibrium estimators D(ij), D2, r(ij), r2 and D(ij) were compared to theoretical distributions. The observed distributions of D(ij), D2, r(ij) and r2 were consistent with the theoretical distribution given an infinite sites model. The observed distribution of D(ij) differed from the theoretical distribution because of an excess of values at -1 and 1. No spatial pattern was observed in the linkage disequilibrium pattern in the Adh region except for two clusters of sites nonrandomly associated in the adult intron and intron 2 of Adh. The magnitude of linkage disequilibrium decreases significantly as nucleotide distance increases, or a distance effect. Adh-Dup had a larger estimate of the recombination parameter, 4Nc, than Adh, where N is the effective population size and c is the recombination rate. A comparison of the mutation and recombination parameters shows that 7-17 recombination events occur for each mutation event. The heterogeneous estimates of the recombination parameter and the inverse relationship between linkage disequilibrium and nucleotide distance are no longer significant when the two clusters of Adh intron sites are excluded from analyses. The most likely explanation for the two clusters of linkage disequilibria is epistatic selection between sites in the cluster to maintain pre-mRNA secondary structure. PMID- 8244014 TI - Cytoplasmic incompatibility and bacterial density in Nasonia vitripennis. AB - Cytoplasmically (maternally) inherited bacteria that cause reproductive incompatibility between strains are widespread among insects. In the parasitoid wasp Nasonia, incompatibility results in improper condensation and fragmentation of the paternal chromosomes in fertilized eggs. Some form of genome imprinting may be involved. Because of haplodiploidy, incompatibility results in conversion of (diploid) female eggs into (haploid) males. Experiments show that bacterial density is correlated with compatibility differences between male and female Nasonia. Males from strains with high bacterial numbers are incompatible with females from strains with lower numbers. Temporal changes in compatibility of females after tetracycline treatment are generally correlated with decreases in bacterial levels in eggs. However, complete loss of bacteria in mature eggs precedes conversion of eggs to the "asymbiont" compatibility type by 3-4 days. This result is consistent with a critical "imprinting" period during egg maturation, when cytoplasmic bacteria determine compatibility. Consequent inheritance of reduced bacterial numbers in F1 progeny has different effects on compatibility type of subsequent male vs. female progeny. In some cases, partial incompatibility occurs which results in reduced offspring numbers, apparently due to incomplete paternal chromosome elimination resulting in aneuploidy. PMID- 8244015 TI - Molecular characterization of a maize B chromosome centric sequence. AB - Supernumerary chromosomes are widespread in the plant kingdom but little is known of their molecular nature or mechanism of origin. We report here the initial cloning of sequences from the maize B chromosome. Our analysis suggests that many sequences are highly repetitive and shared with the normal A chromosomes. However, all clones selected for B-specificity contain at least one copy of a particular repeat. Cytological mapping using B chromosome derivatives and in situ hybridization show that the B specific repeats are derived from the centric region of the chromosome. Sequence analysis of this repeat shows homology to motifs mapped to various plant and animal centromeres and to the maize neocentromere. A precise localization of these sequences among breakpoints within the B centromere and an homology to a facultative centromere, suggest a role for this sequence in centromere function. PMID- 8244016 TI - Simple methods for testing the molecular evolutionary clock hypothesis. AB - Simple statistical methods for testing the molecular evolutionary clock hypothesis are developed which can be applied to both nucleotide and amino acid sequences. These methods are based on the chi-square test and are applicable even when the pattern of substitution rates is unknown and/or the substitution rate varies among different sites. Furthermore, some of the methods can be applied even when the outgroup is unknown. Using computer simulations, these methods were compared with the likelihood ratio test and the relative rate test. The results indicate that the powers of the present methods are similar to those of the likelihood ratio test and the relative rate test, in spite of the fact that the latter two tests assume that the pattern of substitution rates follows a certain model and that the substitution rate is the same among different sites, while such assumptions are not necessary to apply the present methods. Therefore, the present methods might be useful. PMID- 8244017 TI - Stalking the elusive recombination intermediate. PMID- 8244018 TI - Sequence analysis of four new heat-shock genes constituting the hslTS/ibpAB and hslVU operons in Escherichia coli. AB - Sequences of four new heat-shock (HS) genes of Escherichia coli organized into two operons were determined. The operon at 83 min specifies two proteins of 15.8 kDa (HslT) and 16.1 kDa (HslS), which are identical to IbpA and IbpB, respectively. Expression of mRNA from a sigma 32-dependent promoter of the hslTS/ibpAB operon is stimulated 30-75-fold upon temperature upshift. The transcription start point (tsp) is located at a G, 96 bp upstream from the AUG start codon of hslT/ibpA. The deduced amino acid sequences of HslT/IbpA and HslS/IbpB are 48% identical to each other and were found to be remotely related to the chloroplast low-molecular-weight HS protein, which is highly conserved among plants. The second hs operon is much less actively stimulated by temperature upshift, although it has a hs promoter that perfectly matches the consensus of promoters recognized by sigma 32. Located at 88.9 min, the hslVU operon specifies proteins of 19.1 kDa (HslV) and 49.6 kDa (HslU). Multiple tsp were found in this operon. HslV is remotely related to the eukaryotic proteasome proteins, and HslU is very similar to a Pasteurella haemolytica protein of unknown function. Both HslU and the P. haemolytica protein share a ATP/GTP binding motif near their N-termini. The two operons described here are transcribed counterclockwise on the standard genetic map. PMID- 8244019 TI - A T7 RNA polymerase-based system for the construction of Pseudomonas strains with phenotypes dependent on TOL-meta pathway effectors. AB - A general method to construct recombinant Pseudomonas putida (and related bacteria), which transcribe specific genes inserted into their chromosome in response to the presence of alkyl- and halobenzoates, has been developed. The system is based on the ability of the T7 RNA polymerase (T7pol) to initiate transcription from cognate promoter sequences located upstream from cloned genes. A specialized transposon, mini-Tn5 xylS/Pm::T7pol, was constructed which contains the structural T7 gene 1 downstream from the XylS protein/benzoate-regulated Pm promoter of the meta-operon of the TOL catabolic plasmid. This transposon was stably inserted into the chromosome of a P. putida target strain so that gene 1 is transcribed upon exposure of the bacteria to benzoate effectors of the XylS regulator. Genes whose expression is to be mediated by T7pol are cloned in mini Tn5 transposons containing T7 promoter sequences upstream from the cloning site and then the hybrid transposons are inserted into different positions in the same chromosome. In this way, expression of the genes cloned within the mini-Tn5 vectors is dependent on the T7pol/XylS/Pm system. This expression device is particularly well suited for applications in which the expression of two or more genes is to take place in response to a single chemical signal, i.e., exposure to certain aromatic compounds. PMID- 8244020 TI - Sequence and arrangement of two genes of the butyrate-synthesis pathway of Clostridium acetobutylicum ATCC 824. AB - The genes encoding both Clostridium acetobutylicum ATCC 824 butyrate synthesis pathway enzymes, phosphotransbutyrylase (ptb) and butyrate kinase (buk), were sequenced. The genes are immediately adjacent on the chromosome, with ptb preceding buk. A single transcription start point (tsp) was identified 57 bp upstream from the ptb start codon by primer extension analysis. The ptb and buk genes appear to form an operon. A putative Rho-independent terminator structure was identified 26 bp downstream from buk. PMID- 8244021 TI - Cloning and sequence analysis of the gene encoding a thermostable chitinase from Streptomyces thermoviolaceus OPC-520. AB - The gene (chi40) encoding a thermostable chitinase from Streptomyces thermoviolaceus OPC-520 was cloned in Escherichia coli JM109 using pUC18. The nucleotide (nt) sequence of chi40 has been determined. A single open reading frame (ORF) encoded a protein consisting of 414 amino acids (aa) with a M(r) of 43,838. The deduced aa sequence of the cloned chitinase (Chi40) showed striking homology (74%) with Chi63 from Streptomyces plicatus. Comparison with other chitinases revealed that Chi40 contained the two conserved regions common to microbial and plant chitinases. Furthermore, the putative promoter region of chi40, which might be involved in the repression and induction of such catabolite controlled genes, was detected by the DNA sequence alignments. PMID- 8244022 TI - Sequence of a fission yeast gene encoding a protein with extensive homology to eukaryotic elongation factor-1 gamma. AB - A polypeptide with an apparent molecular mass of 23 kDa was identified, that exhibited an affinity to a 491-bp DNA derived from one of the Schizosaccharomyces pombe centromeric DNAs (cen1). After determining its N-terminal amino acid (aa) sequence, a Sz. pombe genomic DNA encompassing the coding sequence of the isolated protein was cloned, and a 2.3-kb genomic DNA region sequenced. Further sequence analysis of cDNA clones, originating from this particular genomic region, confirmed the existence of an open reading frame with a short intron, which encodes a 409-aa protein with striking homology to eukaryotic elongation factor-1 gamma. PMID- 8244023 TI - Inactivation of a yeast transactivator by the fused HIV-1 proteinase: a simple assay for inhibitors of the viral enzyme activity. AB - The human immunodeficiency virus type 1 (HIV-1) proteinase (PR) and its flanking sequences have been fused in frame between the DNA-binding domain and the transcription-activation domain of the yeast protein, GAL4. As has been shown before with the 3C proteinase of Coxsackie virus B3 (CVB3) [Das Mahapatra et al., Proc. Natl. Acad. Sci. USA 89 (1992) 4159-4162], the GAL4::PR fusion protein retains its GAL4 function, providing the PR is inactive. When PR is active, its autocatalytic activity in the hybrid protein is shown to inactivate the transactivation function of GAL4. This provides a simple assay to monitor PR activity. A dose-dependent effect of a potent PR-specific inhibitor is demonstrated in this system and illustrates the sensitivity of the assay. The assay is used for high throughput screening to identify novel inhibitors of the viral PR, and provides a method to generate and analyze mutants and revertants of the PR. PMID- 8244024 TI - Chitin synthase-encoding gene(s) of the Zygomycete fungus Phycomyces blakesleeanus. AB - Using polymerase chain reaction (PCR) primers to two highly conserved sequences within fungal chitin synthase (CHS)-encoding genes, an approximately 750-bp DNA fragment was amplified from genomic DNA of Phycomyces blakesleeanus. The amino acid sequence deduced from the nucleotide sequence of this fragment best matches the motifs found in class-II CHS. The fragment includes an approximately 160-bp region that likely is an intron. Southern hybridization of restriction enzyme digested genomic DNA, using the PCR-amplified DNA as the probe, suggests that P. blakesleeanus contains additional CHS-encoding genes (CHS). To our knowledge, this is the first report on the detection of a CHS gene in a Zygomycete fungus. These studies represent a major step toward exploring the molecular mechanisms of CHS regulation in Phycomyces. The prospects are exciting, since CHS is implicated to play a central role in the sensory responses of P. blakesleeanus involving growth modulations. PMID- 8244025 TI - The sequences of gene rIII of bacteriophage T4 and its mutants. AB - The nucleotide sequences of bacteriophage T4 gene rIII, from six different rIII mutants, have been determined. We show that the mutations r67, rES35 and rES40 cause basic amino acid changes in the rIII protein, while the mutations rBB9, rCR28 and rCP24 cause chain termination. PMID- 8244026 TI - Sequence of the triosephosphate isomerase-encoding gene isolated from the thermophile Bacillus stearothermophilus. AB - By analysis of genomic clones, we have determined the complete nucleotide sequence of the gene encoding triosephosphate isomerase (TIM; EC 5.3.1.1) in the thermophilic bacterium, Bacillus stearothermophilus. The gene encodes a 253-amino acid TIM which is 39% identical to that of the mesophile, Escherichia coli. PMID- 8244027 TI - Characterization of a Streptomyces antibioticus gene cluster encoding a glycosyltransferase involved in oleandomycin inactivation. AB - By homology to the mgt gene (encoding a macrolide glycosyltransferase) from Streptomyces lividans, a 3.3-kb DNA fragment from the oleandomycin producer, Streptomyces antibioticus, was cloned and sequenced. Analysis of the sequence revealed the presence of the 3' end of a gene (ORF1) and two complete ORFs (ORF2 and oleD), all of them translationally coupled. The deduced product of the sequenced region of ORF1 contained the typical signature of integral membrane proteins responsible for the translocation of substrates across the membrane. The ORF2 product did not show significant similarity with proteins in databases, but contains an N-terminus leader peptide region characteristic of secreted proteins, and a lipid attachment site motif characteristic of membrane lipoproteins synthesized with a precursor signal peptide. The oleD product showed clear similarity with several UDP-glucuronosyl- and UDP-glycosyl-transferases from different origins and particularly with the mgt gene from S. lividans, and might encode a glycosyltransferase activity capable of inactivating macrolides. It is proposed that these three genes could participate in the intracellular glycosylation of oleandomycin and its secretion during antibiotic production. PMID- 8244028 TI - Codon preference in corynebacteria. AB - The codon usage (CU) of 34 genes from the closely related species, Brevibacterium lactofermentum and Corynebacterium glutamicum (BLCG), was analysed and compared with that of 23 genes from other Brevibacterium and Corynebacterium species. The G+C content of the BLCG genes ranged from 50 to 62%. A wider range was found in other corynebacterial genes (25-71%). The G+C contents of non-coding regions in glutamic acid bacteria are lower than those of the coding regions and both values are lower than the G+C content of ribosomal RNA (rRNA) sequences, suggesting an unusual biased mutation pressure. The CU and synonymous codon usage (SCU) analysis showed several common characteristics among the sequenced corynebacterial genes, consistent with the close relatedness of B. lactofermentum and C. glutamicum. A subset of 25 preferred codons were deduced from the presumably highly expressed genes and they encode most of the amino acid (aa) residues of the BLCG group. An analysis of the effective number of codons (Nc) was carried out in order to check the GC3s (G+C content at the silent third position of sense codons) dependence of the CU in corynebacteria. Nc values showed differences between the BLCG group and other corynebacterial sequences. A comparison of the most used codons for each aa showed a stronger similarity to Streptomyces than to Escherichia coli. The CU/SCU tables of corynebacteria are useful for identification of protein-coding regions, including start codons when they are uncertain, and for designing oligodeoxyribonucleotide probes from an aa sequence. PMID- 8244029 TI - Multiple tandem integrations of transforming DNA sequences in yeast chromosomes suggest a mechanism for integrative transformation by homologous recombination. AB - In yeast, the fate of linear DNA molecules upon transformation is determined by the existence of sequence homology between chromosomes and the ends of the transforming molecule. To understand the mechanism of integration of transforming DNA, we have studied the influence of DNA concentration on the frequency and type of transformants obtained, using either non-replicative or replicative plasmids. In both cases, increasing DNA concentration results in multiple tandem repeats integrated into the chromosome containing the homologous target sequence. When a diploid strain is transformed, multiple tandem repeats occur in only one of the two homologous chromosomes at a time. The frequency distribution of the different types of integrants observed indicates non-independent integration events likely to result from plasmid-plasmid interaction prior to chromosome integration. In addition, our results define the proper conditions for optimized gene targetting or gene rescue experiments. PMID- 8244030 TI - A Saccharomyces cerevisiae protein-serine kinase related to mammalian glycogen synthase kinase-3 and the Drosophila melanogaster gene shaggy product. AB - The glycogen synthase kinase-3 (GSK-3) family of protein-serine kinases is implicated in the development and hormonal regulation of higher eukaryotes. GSK-3 related genes have been cloned and characterized in mammals (alpha and beta forms), Drosophila melanogaster (shaggy/zeste-white3) and Saccharomyces cerevisiae (MCK1). Using the polymerase chain reaction and primers designed to hybridize to conserved catalytic domain sequences of this family, a genomic fragment was amplified from budding yeast DNA. Genomic clones encompassing the entire reading frame were subsequently isolated and sequenced. The protein encoded by this gene, termed ScGSK-3, displays high identity with members of the GSK-3 family, sharing several structural features including a regulatory Tyr residue. A phylogenetic analysis of the catalytic domains of these protein kinases suggests that ScGSK-3 represents the bona fide homologue of GSK-3 and the shaggy product, while the related MCK1 protein kinase is encoded by a paralogous gene which originated by a gene duplication event in the yeast lineage. PMID- 8244031 TI - The functional analysis of nonsense suppressors derived from in vitro engineered Saccharomyces cerevisiae tRNA(Trp) genes. AB - Nonsense suppressors derived from Saccharomyces cerevisiae tRNA(Trp) genes have not been identified by classical genetic screens, although one can construct efficient amber (am) suppressors from them by making the appropriate anticodon mutation in vitro. Herein, a series of in vitro constructed putative suppressor genes was produced to test if pre-tRNA(Trp) processing difficulties could help to explain the lack of classical tRNA(Trp)-based suppressors. It is clear that inefficient processing of introns from precursor tRNA(Trp), or inaccurate overall processing, may explain why some of these constructs fail to promote nonsense suppression in vivo. However, deficient processing must be only one of the reasons why classical tRNA(Trp)-based suppressors have not been characterized, as suppression may still be extremely weak or absent in instances where the in vitro construct can lead to an accumulation of mature tRNA(Trp). Furthermore, suppression is also very weak in strains transformed with an intronless derivative of a putative tRNA(Trp) ochre (oc) suppressor gene, wherein intron removal cannot pose a problem. PMID- 8244032 TI - Small subunit rDNA variation in a population of lichen fungi due to optional group-I introns. AB - A natural population of the lichen-forming ascomycetous fungus, Cladonia chlorophaea, contained individuals with small subunit ribosomal DNA (SSU rDNA) of at least four different size classes and nine restriction-site patterns. The source of these differences was the variable occurrence of 200-400-nucleotide insertions, previously identified as small group-I introns, at five different positions within the SSU coding region. By specific amplification of the sequences flanking these five intron positions with the polymerase chain reaction (PCR), a minimum of nine types of rDNA repeats were defined that differ in number, position, restriction pattern and size of introns. The positions of the introns were verified by sequence analysis. The variable distribution of these introns suggests that they are currently mobile--either by intron insertion, deletion or both--within this species complex. PMID- 8244033 TI - Recombinant DNA procedures for producing small antimicrobial cationic peptides in bacteria. AB - Natural polycationic antibiotic peptides have been found in many different species of animals and insects and shown to have broad antimicrobial activity. To permit further studies on these peptides, bacterial expression systems were developed. Attempts to produce these peptides with an N-terminal signal sequence were unsuccessful due to the lability of the basic peptides. Therefore, a number of different fusion protein systems were tested, including fusions to glutathione S-transferase (GST) (on plasmid pGEX-KP), Pseudomonas aeruginosa outer membrane protein OprF (on plasmid pRW5), Staphylococcus aureus protein A (on plasmid pRIT5), and the duplicated IgG-binding domains of protein A (on plasmid pEZZ18). In the first three cases, stable fusion proteins with the defensin, human neutrophil peptide 1 (HNP-1), and/or a synthetic cecropin/melittin hybrid (CEME) were obtained. In the course of these studies, we developed a novel method of purifying inclusion bodies, using the detergent octyl-polyoxyethylene (octyl POE), as well as establishing methods for preventing fusion protein proteolytic breakdown. Cationic peptides could be successfully released from the carrier protein with high efficiency by chemical means (CNBr cleavage) and with low efficiency by enzymatic cleavage (using factor Xa protease). Fusions of protein A to cationic peptides were secreted into the culture supernatant of S. aureus clones and after affinity purification, CNBr digestion and column chromatography, pure cationic peptide was obtained. CEME produced by this procedure had the same amino acid (aa) content, aa sequence, gel electrophoretic mobility and antibacterial activity as CEME produced by protein chemical procedures. PMID- 8244034 TI - Recombinant Leishmania surface glycoprotein GP63 is secreted in the baculovirus expression system as a latent metalloproteinase. AB - A gene encoding the Leishmania surface metalloproteinase, GP63, was modified using the polymerase chain reaction to obtain effective secretion of recombinant GP63 (reGP63) in the baculovirus insect cell expression system. The coding region for the N-terminal signal peptide (SP) of GP63 was modified to resemble the SP for the GP67 envelope protein from the budded virus form of Autographa californica nuclear polyhedrosis virus. To prevent processing at the C-terminus with a glycosyl phosphatidylinositol anchor and the subsequent membrane anchoring of reGP63 in insect cells, the coding region for a putative SP at the C-terminus of GP63 was deleted. The reGP63 protein was glycosylated and secreted as a latent metalloproteinase in the baculovirus expression system. The reGP63 protein was purified from serum-free medium using concanavalin A lectin affinity chromatography, with a yield of 1 mg/l. The purified Leishmania reGP63 was secreted as a latent proteinase. Treatment of reGP63 with HgCl2 resulted in activation of full proteinase activity and a concomitant decrease in M(r). The mechanism of the activation of Leishmania reGP63 is consistent with that of other members of the family of matrix-degrading metalloproteinases. PMID- 8244035 TI - Generation of a plasmid vector for deletion cloning by rapid multiple site directed mutagenesis. AB - The construction of a new plasmid vector, devoid of all MboI (GATC) and TspEI (AATT) restriction sites, is described. The lack of these two frequent-cutting restriction sites is a unique feature among plasmids. This new plasmid, pBRkanf1 , allows selective fragmentation of a cloned insert. As a result, the vector offers an alternative strategy to create overlapping and sequentially deleted subclones. In addition, the construction of the new plasmid required the development of a rapid and accurate multiple site-directed mutagenesis procedure. The mutagenesis method uses a combination of DNA amplification and chain extension by DNA polymerase. By this method, mutations are created progressively from one end of a DNA molecule to the other. PMID- 8244036 TI - Two plasmids, X1918 and Z1918, for easy recovery of the xylE and lacZ reporter genes. AB - Two plasmids, X1918 and Z1918, were constructed which contain the promoter-less xylE and lacZ reporter genes flanked symmetrically by the multiple cloning site (MCS) from pUC19. These cassettes can easily be derived from the multicopy plasmid, pUC1918. PMID- 8244037 TI - Sequence and structure of cmp, the replication enhancer of the Staphylococcus aureus plasmid pT181. AB - The Staphylococcus aureus plasmid pT181 possesses a DNA replication enhancer element, called cmp, that is required in cis for optimal utilization of the initiator protein by the origin of replication. The minimal nucleotide sequence required for cmp activity was defined by testing progressively smaller DNA fragments for their ability to restore cmp activity in a plasmid mutant deleted for cmp. These experiments indicate that cmp is a sequence of 100 base pairs (bp) characterized by a loosely repeated sequence motif and phased oligo(dT) tracts. Intrinsic DNA bending at cmp was detected by a circular permutation assay of the locus using polyacrylamide-gel electrophoresis and by computer modeling. The cmp element was found to contain two loci of intrinsically bent DNA that confer an overall bent conformation to this replication enhancer. PMID- 8244038 TI - Sequence and characterization of mutT from Proteus vulgaris. AB - A cloned DNA fragment containing the tryptophanase (tna) operon of Proteus vulgaris was found to contain a gene analogous to mutT of Escherichia coli immediately distal to the tna operon. The presumptive mutT of P. vulgaris was shown to be a functional gene by complementation of a mutT mutant from E. coli. The deduced amino acid sequence of the MutT polypeptide of P. vulgaris was 47% identical and 70% similar to MutT of E. coli. The mutT and tna operons of P. vulgaris were shown to be adjacent on the genome of this organism. These operons are located about 20 min apart in the E. coli genome. Our findings suggest that either or both tna and mutT have different genomic locations in the two organisms. PMID- 8244039 TI - Safety and efficacy of paroxetine in elderly patients. PMID- 8244040 TI - Depression in the elderly: when to treat, when to refer. PMID- 8244041 TI - Prevalence and consequences of depression in the elderly. PMID- 8244042 TI - Tricyclic antidepressants: advantages and limitations. PMID- 8244043 TI - Using SSRIs to treat depression in the elderly. PMID- 8244044 TI - Association of autoimmunity to vascular heparan sulfate proteoglycan and vascular disease in the aged. AB - In the very old, there is an increased prevalence of immunologically mediated vascular disease. Vascular heparan sulfate proteoglycan (vHSPG) is a molecule which plays an important structural and functional role in the vasculature. Autoimmunity to vHSPG may play a role in the pathogenesis of vascular disease. The presence of autoantibodies to vHSPG was investigated in sera from aged nursing home patients who have a high prevalence of vascular disease. The results showed an association of the presence of autoantibodies to vHSPG in the sera from patients in this aged population with vascular disease, particularly cerebrovascular disease, renal disease and diabetes. In addition, autoantibodies to vHSPG protein core were associated with cerebrovascular disease in this frail elderly population. These studies support the hypothesis that autoimmunity may play a role in vascular disease in the aged. PMID- 8244045 TI - Age-induced nerve cell loss in the myenteric plexus of the small intestine in man. AB - We examined the number of nerve cells of the myenteric plexus and the thickness of the smooth muscle in the small intestine in autopsy material of 6 young and 6 old persons. Neurons in the myenteric plexus have been visualised by a nonhistochemical method (Giemsa) in laminar preparations of the muscularis externa. Significant reductions of at least 34% in the number of neurons in the ganglia of the myenteric plexus of the old subjects were recorded in all regions of the small intestine, especially in the duodenum where the number of neurons decreased by over 38%. However, there was no significant correlation between nerve cell count and thickness of intestinal smooth muscle since no difference was found in thickness of intestinal smooth muscle between young and old subjects. The decrease in the neuron density with age could affect the potential of the enteric nervous system to influence control over several small intestinal functional parameters. PMID- 8244046 TI - Decreased melatonin biosynthesis, calcium flux, pineal gland calcification and aging: a hypothetical framework. AB - Increased pineal calcifications and decreased pineal melatonin biosynthesis, both age related, support the notion of a pineal bio-organic timing mechanism. Decreased calcium ion availability is the single common denominator of diminished beta-postreceptor- and alpha-receptor-stimulating functions in beta-receptor potentiation, which is necessary for nocturnal peak melatonin production. A comprehensive framework for the interaction of aging pineal cell mechanisms, calcium flux and melatonin biosynthesis is presented. PMID- 8244047 TI - Decreased peripheral glucocorticoid sensitivity in Alzheimer's disease. AB - Both peripheral and central glucocorticoid sensitivity was examined in patients with Alzheimer's disease (AD; n = 13), glucocorticoid-treated patients (n = 8), healthy elderly controls (n = 10) and young controls (n = 9). We performed glucocorticoid receptor-mediated skin vasoconstrictor responses to clobetasol and low-dose dexamethasone suppression tests. Patients with AD showed skin blanching at a significantly higher clobetasol concentration than did healthy elderly controls (p = 0.002). There was no difference in skin blanching between patients with AD and patients treated with corticosteroids. Patients with AD had significantly higher post-dexamethasone serum cortisol levels than healthy elderly (p = 0.01). No association was found between skin blanching and dexamethasone suppressibility. Thus patients with AD have apparently independent reductions in both central nervous system and peripheral glucocorticoid sensitivity. These results predict an increase in glucocorticoid secretion in some patients, which might accelerate neuronal degeneration in the absence of features of overexposure to glucocorticoids in peripheral tissues. PMID- 8244048 TI - Isolated systolic hypertension in the elderly: survey of practitioners' attitude and management. AB - A postal survey of 445 medical practitioners was carried out in the light of recent large studies supporting the treatment of isolated systolic hypertension (ISH) in the elderly. The response rate was 60.2%. 46% of the respondents would consider using drug therapy for ISH, and this was dependent on the patients age and the level of systolic blood pressure. Thiazide diuretic was the most popular first choice drug. The most recurring reason for not using drug therapy by 54% of medical practitioners was that treatment with drugs will reduce quality of life due to side effects of drugs. There was a large consensus on the use of non pharmacological treatment of ISH in the elderly. PMID- 8244049 TI - Plasma DNA as cell death marker in elderly patients. AB - Plasma DNA increases where cell death occurs in vivo. To investigate its significance in elderly patients, plasma DNA was assayed in 79 institutionalized patients over 68 years of age. The patients were divided into two groups: group I comprises 39 patients suffering from various acute or chronic illnesses; group II comprises 40 patients without chronic disease, and free of any clinical or biological symptoms of any infectious or inflammatory process. Plasma DNA was higher in group I than in group II (p < 0.0001) and in group II than in a control group of middle-aged subjects (p < 0.05). In group I, increase in plasma DNA concentration was found in various pathological situations associated with cell death phenomena, including infections, cancers with metastasis, hepatitis, irreversible cardiac failure, severe respiratory insufficiency and thrombophlebitis. Plasma DNA concentrations were not correlated with erythrocyte sedimentation rate, fibrinogen concentration, hemoglobin concentration or leukocyte count. In group I, as well as in the overall population, survival after 1 month was significantly reduced in patients with increased concentrations of plasma DNA. In conclusion, plasma DNA as a marker of cell death phenomena occurring in vivo, could be helpful for follow-up and management of elderly patients. PMID- 8244050 TI - Characteristics of colorectal cancer in elderly patients. AB - The aim of the study was to examine the characteristics of colorectal cancer in elderly patients. The medical histories of 178 consecutive patients (79 men and 99 women) with colorectal cancer were studied from hospital records. The studied parameters were compared between three age groups: under 65 years old, 65-80 years old and over 80 years old. In the two older age groups, cancer was situated more commonly in the proximal colon than in the youngest age group (p = 0.029). Especially in elderly subjects, rectosigmoidoscopy is not sufficient since up to 40% of cancers are located in the right colon. Colonoscopy is recommended as the method of choice in old patients if colorectal cancer is suspected. Cancers in the oldest age group more often fell into to Dukes' stages C1, C2 and D (metastatic growth) (p = 0.072) and they were greater in size than those of patients under 80 years old (p = 0.029). Small cancers (under 5 cm) had obvious blood in the stools as the main symptom and represented nonmetastatic Dukes' stages. If these cancers could be detected earlier by testing for occult blood, the prognosis of colorectal cancer would improve. The resectability of cancer was lower in patients over 80 years than in other age groups (p = 0.016). However, in logistic regression analysis, the large size, poor or moderate cell differentiation, and distal location of the cancer, but not age, were associated with poor resectability. PMID- 8244051 TI - Parkinson's dementia and Alzheimer's dementia: an evoked potential comparison. AB - Auditory endogenous event-related potentials (ERPs) and flash visual evoked potentials (VEPs) were recorded in 26 elderly patients with idiopathic Parkinson's disease (PD), 14 with dementia and 12 non-demented, 16 elderly patients with Alzheimer dementia (AD) and 15 cognitively intact controls. ERP P3 and flash-VEP N2, P2 and delta (P2-P1) latency measures were significantly increased in the demented PD group compared with controls. The ERP P3 latency was also significantly delayed in the AD group compared with controls, but the differences in the flash-VEP measures from controls were not significant. No significant differences were noted between the PD groups, except for a significantly shorter flash-VEP N1 latency in the demented PD group; this was also the only significant evoked potential difference between the AD and PD dementia groups, which were otherwise electrophysiologically similar. PMID- 8244052 TI - [Structure and sources of river and drinking water pollution by volatile organochlorine compounds in the Kuzbass region]. AB - Chemical pollution of the river and drinking water with volatile organochlorine compounds in the Kuznetsk coal fields was noted. Necessity of the alternative to chlorination way of water disinfection was discussed. PMID- 8244053 TI - [Environmental protection from pollution with agricultural toxic substances in the Vinnitsa region]. PMID- 8244054 TI - [Evaluation of Ricide-P pesticide toxicity using the experimental model mother fetus-newborn]. AB - Necessity of the mother-fetus-newborn model usage in toxicological experiments for the xenobiotics environmental content standardization was shown in the toxicological study of Ricid-P pesticide. PMID- 8244055 TI - [Pesticide standardization in the soil as a method of environmental quality regulation]. PMID- 8244056 TI - [Sanitary and toxicologic characteristics of anisaldehyde]. AB - The threshold dose of anisealdehyde was ascertained at the level of 1,8 mg/kg, and maximum noneffective one - 0,04 mg/kg. PMID- 8244057 TI - [Ecologic and hygienic evaluation of soil of industrial wasteland non-ferrous metallurgy and recycling plants]. PMID- 8244058 TI - [Current problems of drinking water fluoridation (Review)]. PMID- 8244059 TI - [Environmental protection and the population's health status with reutilization of sewage in foreign countries (Review)]. PMID- 8244060 TI - [Hygienic principles in evaluation and risk management in occupational diseases]. PMID- 8244061 TI - [Hygienic evaluation of workplace environment in the wool processing industry]. PMID- 8244062 TI - [Experimental evaluation of fibrogenic action of asbestos dust]. PMID- 8244063 TI - [Information for assessment of maximum permissible concentration of fire extinguishing powders of the pirant group in the workplace]. PMID- 8244064 TI - [Correlation between environmental pollution and morbidity level in urban children]. PMID- 8244065 TI - [Hygienic characteristics of dust and gas waste mixtures in ship scrap cutting]. PMID- 8244066 TI - [Physiologic and hygienic evaluation of working conditions and effectiveness of individual protective devices in sand-blast work]. PMID- 8244067 TI - [Introduction of research results on children and adolescents hygiene into sanitary and epidemiologic managerial practice]. AB - Most important results of investigations at Research Institute of Hygiene and Prevention of Diseases of Children, Adolescents, and Youth and at other analogous institutions are given. Standards and methodological documents in this sphere are mentioned. PMID- 8244068 TI - [Professional orientation of youth in foreign countries (Review)]. PMID- 8244069 TI - [Medical and biological research on Moscow children's health status in districts with various levels of air pollution]. PMID- 8244070 TI - [Ceruloplasmin as an indicator of biological activity of resin and it's ingredients]. PMID- 8244071 TI - [Utilization of chromatographic parameters for prognosis of toxicity of hazardous substances and their fate in the biosphere]. PMID- 8244072 TI - [Air environment in short-term recreation areas of industrial cities in Uzbekistan]. PMID- 8244073 TI - [Ecologic and hygienic evaluation of industrial waste from non-ferrous alloy plants]. PMID- 8244074 TI - [Spreading of Pseudomonas spp resistant to disinfectants]. PMID- 8244075 TI - [Toxic dynamic characteristics of citric acid with various routes of administration]. PMID- 8244076 TI - [Evaluation of combined action of kharelly preparation components on the reproductive function of experimental animals]. PMID- 8244077 TI - [Toxicologic and hygienic evaluation of some organosilicone compounds]. PMID- 8244078 TI - [Ecologic and hygienic problems of aluminum factories (Review)]. PMID- 8244079 TI - [Lead in animals and man (review)]. PMID- 8244080 TI - [Mass-gas chromatographic spectrometry of organic substances in environmental objects]. PMID- 8244081 TI - [Determination of tylosin in the water supply using a chromatographic method]. PMID- 8244082 TI - [Modeling of dose-response effect of beryllium exposure on the organism by aerogenic inhalation in industrial conditions]. PMID- 8244083 TI - [Gas-chromatographic determination of reiser in environmental objects]. PMID- 8244084 TI - Free radicals in disease processes: a compilation of cause and consequence. PMID- 8244085 TI - Alpha, mu and pi class glutathione S-transferases in human synovium and cultured synovial fibroblasts: effects of interleukin-1 alpha, hydrogen peroxide and inhibition of eicosanoid synthesis. AB - We describe expression of alpha, mu and pi class glutathione S-transferase (GST) and, CuZn- and Mn superoxide dismutase (SOD) in human synovium and cultured synovial fibroblasts. Immunohistochemical and immunoblotting studies showed synovium and cultured cells expressed pi GST and both isoforms of SOD. Cellular localisation was largely perinuclear. No expression of alpha or mu GST was detected even though polymerase chain reaction analysis showed 4/6 subjects had positive genotypes at the polymorphic, mu class GSTM1 locus. Incubation of cultured synovial fibroblasts with H2O2, IL-1 alpha and the cyclooxygenase and lipoxygenase inhibitor, Tenidap, did not induce expression of alpha, mu or pi GST though treatment with IL-1 alpha caused a marked increase in the expression of Mn SOD. PMID- 8244086 TI - Angiotensin converting enzyme inhibitors as oxygen free radical scavengers. AB - The authors have compared the ability of two non-SH-containing angiotensin converting enzyme (ACE) inhibitors (enalaprilat and lisinopril) with an -SH containing ACE inhibitor (captopril) to scavenge the hydroxyl radical (.OH). All three compounds were able to scavenge .OH radicals generated in free solution at approximately diffusion-controlled rates (10(10) M-1 s-1) as established by the deoxyribose assay in the presence of EDTA. The compounds also inhibited deoxyribose degradation in reaction mixtures which did not contain EDTA but not so effectively. This later findings also suggests that they have some degree of metal-binding capability. Chemiluminescence assays of oxidation of hypoxanthine by xanthine oxidase in the presence of luminol, confirm that the three ACE inhibitors are oxygen free radical scavengers. Our results indicate that the presence of a sulphydryl group in the chemical structure of ACE inhibitors is not relevant for their oxygen free radical scavenging ability. PMID- 8244087 TI - Protection of heme proteins by vitamin E, selenium, and beta-carotene against oxidative damage in rat heart, kidney, lung and spleen. AB - Effects of the combination of vitamin E, selenium, and beta-carotene on oxidative damage to rat heart, kidney, lung, and spleen were studied by measurement of the production of oxidized heme proteins (OHP) during spontaneous and prooxidant induced oxidation. Male SD rats were fed with a vitamin E and selenium deficient diet or a diet supplemented with vitamin E, selenium, and beta-carotene. Homogenates of heart, kidney, lung, and spleen were incubated at 37 degrees C with and without the presence of bromotrichloromethane (CBrCl3). The diet supplemented with antioxidants showed a strong protective effect against oxidative damage to heme proteins during the early stages of both spontaneous and CBrCl3-induced oxidation in contrast to the antioxidant deficient diet. Synergism of multiple antioxygenic nutrients against oxidative damage to various animal tissues is discussed. PMID- 8244088 TI - Changes in plasma antioxidant status during eccentric exercise and the effect of vitamin supplementation. AB - Twenty-four healthy students undertook one hour of box-stepping exercise. Prior to exercise eight had received no medication (Group A), eight received 400 mg of vitamin C daily for three weeks before and one week after exercise (Group C) and eight received 400 mg of vitamin E for the same period (Group E). Groups C and E had significantly higher levels of vitamin C (p < 0.01) and vitamin E (p < 0.01) respectively than group A at the commencement of exercise. Plasma total antioxidant capacity rose significantly during exercise in all group (A - p < 0.05; C - p < 0.001; E - p < 0.001). This rise was accounted for by increases in plasma uric acid in all groups. In addition there were significant increases in vitamin C in group C (p < 0.001) and vitamin E in group E (p < 0.05). There were no significant changes in plasma malondialdehyde following exercise in any group. It is concluded that plasma antioxidant capacity rises in response to one hour of eccentric exercise and that the contribution of individual antioxidants to this change can be influenced by vitamin supplementation. The possible mechanisms of the antioxidant changes during exercise and their implications are discussed. PMID- 8244089 TI - Cellularly generated active oxygen species and HeLa cell proliferation. AB - In HeLa cells evidence is provided that active oxygen species such as hydrogen peroxide and superoxide at low levels are important growth regulatory signals. They may constitute a novel regulatory redox system of control superimposed upon the established cell growth signal transduction pathways. Whilst for example hydrogen peroxide can be added exogenously to elicit growth responses in these cells, it is clear that cellularly generated superoxide and hydrogen peroxide are important. Experiments with superoxide dismutase, superoxide dismutase mimics and inhibitors of both superoxide dismutase and xanthine oxidase suggest that superoxide generated intracellularly and superoxide released extracellularly are both relevant to growth control in HeLa cells. PMID- 8244090 TI - A perspective on iron deficiency anaemia. PMID- 8244091 TI - Duodenal ulcer pain--the role of acid and inflammation. PMID- 8244092 TI - Influence of dietary fat on duodenal resistance to acid. AB - Three experimental diets were prepared from a standard formula plus pure oleic, linoleic, or eicosapentaenoic acid (2% by weight). Mucosal resistance to acid was tested in anaesthetised rats fed the experimental diets for at least four weeks (60 rats per diet) by duodenal infusion of HCl (200 to 700 mumol) 30 minutes after pretreatment with either saline or 100 mumol HCl (used as a mild irritant). Rats were killed one hour after the test and the duodenal damage was assessed 'blindly' using a combined macroscopic and histological score. Differences were tested by analysis of covariance of the dose-response curves. Mucosal resistance was similar in the three groups when the acid challenge was given after saline pretreatment. Resistance to acid in all three groups was significantly increased by previous exposure to 100 mumol HCl (p < 0.01). Interestingly, rats fed a linoleic or eicosapentaenoic supplemented diet after pretreatment with HCl developed significantly higher resistance to acid than those fed the diet with oleic acid (p < 0.05). Pretreatment with indomethacin suppressed the difference between diets. In conclusion, dietary polyunsaturated fatty acids enhance duodenal resistance to acid by potentiation of adaptive cytoprotection. PMID- 8244093 TI - Reduced tissue type plasminogen activator activity of the gastroduodenal mucosa in peptic ulcer disease. AB - The gastroduodenal mucosa has a rich blood supply. An active fibrinolytic system is presumably required to maintain vascular patency, and impairment may result in reduced blood flow, focal tissue necrosis, and peptic ulcerogenesis. Tissue type and urokinase type plasminogen activator activity (expressed as mIU/mg protein) and plasminogen activator inhibitor type-1 antigen were assayed in homogenates of gastric and duodenal biopsy specimens taken from patients with: normal endoscopy (controls) (n = 14); active duodenal ulcer (n = 21); healed duodenal ulcer (n = 12); and active benign gastric ulcer (n = 15). In controls mean duodenal tissue type plasminogen activator activity was 4110 and urokinase type plasminogen activator activity 150; gastric tissue type plasminogen activator was 2760 and urokinase type plasminogen activator 170; plasminogen activator inhibitor type-1 was generally undetectable. At the edge of active duodenal ulcers tissue type plasminogen activator was considerably reduced, 2220 (p < 0.001) whereas urokinase type plasminogen activator was raised, 290 (p < 0.01). At the edge of active benign gastric ulcers tissue type plasminogen activator was substantially reduced, 1160 (p < 0.001) but urokinase type plasminogen activator was unchanged. At the scar of healed duodenal ulcers tissue type plasminogen activator was slightly reduced, 3290, but urokinase type plasminogen activator was increased, 308 (p < 0.05). H2 receptor antagonist treatment had little effect on tissue type or urokinase type plasminogen activator activity. Plasminogen activator inhibitor type-1 was increased at the edge of active ulcers (p < 0.05) especially when tissue type plasminogen activity was low (r = -0.61, p < 0.05). These findings are consistent with the hypothesis that impaired fibrinolytic activity may be implicated in peptic ulcerogenesis. PMID- 8244094 TI - Diagnostic value of serum pepsinogen C in patients with raised serum concentrations of pepsinogen A. AB - Hypopepsinogenaemia A is often found in patients with gastric atrophy and gastric surgery. In these conditions serum pepsinogen C provides additional diagnostic information, especially when expressed as pepsinogen A:C ratio. Hyperpepsinogenaemia A has been shown in patients with duodenal ulcer disease, Zollinger-Ellison syndrome, hypertrophic gastropathy, chronic renal failure, and during omeprazole treatment. As patients with hyperpepsinogenaemia A often overlap in symptoms, endoscopical findings, and serum gastrin values, this study has evaluated whether measurement of serum pepsinogen C in subjects with hyperpepsinogenaemia A can help in differentiating clinical conditions. Serum concentrations of pepsinogen A and C were measured in serologically Helicobacter pylori negative blood transfusion donors (127) as reference population, and in patients with Zollinger-Ellison syndrome (24), duodenal ulcer (50), hypertrophic gastropathy (5), and chronic renal failure (50), and also in reflux oesophagitis patients on longterm omeprazole treatment (28). A low pepsinogen A:C ratio was found in all patients with hypertrophic gastropathy. A pepsinogen A:C ratio above the critical value of 4.7 was found in 14 (70.0%) of the Zollinger-Ellison patients, two (9.5%) of the duodenal ulcer patients, 11 (25.6%) of the patients with chronic renal failure, and in one (7.1%) of the patients receiving longterm omeprazole treatment. In fact, all but three hyperpepsinogenaemia A patients with a pepsinogen A:C ratio greater than 4.7 and normal renal function had the Zollinger-Ellison syndrome. In patients with hyperpepsinogenaemia A, a low pepsinogen A:C ratio may point to hypertrophic gastropathy, while a pepsinogen A:C ratio greater than 4.7 is suggestive for the Zollinger-Ellison syndrome. PMID- 8244095 TI - Prospective multicentre study of risk factors associated with delayed healing of recurrent duodenal ulcers (RUDER). RUDER Study Group. AB - Risk factors for delayed duodenal ulcer healing during treatment with ranitidine (300 mg daily) were examined in a multicentre German study of 1923 patients with endoscopically proved, recurrent duodenal ulceration. Healing rates, per protocol, were 39.5% at two weeks, 70.9% at four weeks, and 93.2% at eight weeks. Prospective testing of five, predefined risk factors indicated that smoking (p = 0.0039) was associated with a decreased healing rate at two weeks. Frequent prior recurrence (p = 0.464), a heavy physical workload (p = 0.145), and psychological stress (p = 0.062) were not associated with a decreased healing rate and there were too few patients at risk to allow assessment of the effect of regular NSAID intake. Exploratory analysis identified prior slow healing, a large ulcer, multiple ulcers, and prior ulcer complications, in addition to smoking, as markers of slow healing. In the absence of these risk factors, the mean healing time was 3.3 weeks (95% confidence interval 3.0, 3.5), rising to 3.7 weeks (3.5, 3.9) for one, 4.4 weeks (4.1, 4.7) for two, and 5.1 weeks (4.5, 5.6) for three to five risk factors. Delayed duodenal ulcer healing is associated with multiple factors whose effect is cumulative; for patients with two or more of five easily identified risk factors, more than four weeks' treatment with a histamine H2 receptor antagonist is required to achieve ulcer healing. PMID- 8244096 TI - Post-bulbar and coexisting ulceration: unique features of peptic ulcer in Hyderabad. AB - Post-bulbar ulceration is uncommon, but a pilot study in Hyderabad showed a high incidence. We therefore carried out a prospective endoscopic study of the distribution of peptic ulceration and its relation to symptoms and demography. Of the 360 consecutive patients referred for endoscopy, 113 (92 men, 21 women) had peptic ulceration. Median age 35 years, median duration one year. Five patients (4%) had gastric ulcer, 77 (68%) had duodenal ulcer, and 31 (28%) had coexisting gastric and duodenal ulcer. The duodenal ulcer was found in the pyloric canal in 14% of patients, in the bulb in 80%, and the post-bulbar region in 56% of patients. Sixty seven per cent of duodenal ulcers were located at more than one site. The incidence of post-bulbar v bulbar ulcer was 1:1.5. Deformed bulb was seen in 50% of duodenal ulcer patients, but haemorrhage and stenosis were uncommon. Except for nocturnal pain, there were no differences in symptoms between the groups. Forty two per cent of patients smoked, 15% chewed tobacco, and 18% drank alcohol; almost all were men. Sixty four per cent drank tea. The staple diet (85%) was rice, and 70% used tamarind and spices daily. Duodenal ulcer was three times more common than gastric ulcer with a high incidence of post-bulbar and coexisting ulcer. It affected a younger and predominantly male population, and was not associated with a higher rate of complication. PMID- 8244097 TI - Properties of a potassium channel in cultured human gastric cells (HGT-1) possessing specific omeprazole binding sites. AB - The HGT-1 human gastric cell line is similar to acid secreting parietal cells in that it possesses H2 receptors, histamine sensitive adenyl cyclase, and Cl- channels, which are activated by histamine by a cyclic adenosine monophosphate (cAMP) dependent mechanism. To discover if HGT-1 cells have additional properties found in parietal cells, [3H]omeprazole and patch clamp recording techniques were used to evaluate specific omeprazole binding sites and K+ channels in the plasma membrane. HGT-1 cells exhibited [3H]omeprazole binding in the non-stimulated state, which increased 100% in the presence of 1 mM histamine. High conductance (about 155 pS) K+ channels were active spontaneously in 17% of cell attached or excised inside out patches in non-stimulated subconfluent HGT-1 cells. In inside out patches, channel activity increased fivefold during depolarisation, ion substitution experiments confirmed that the channels were highly selective for K+, and channel activity was almost abolished by removal of Ca2+ or addition of 5 mM Ba2+. In quiescent cell attached patches, 0.1 mM dibutyryl cAMP failed to activate K+ channels. In contrast, 6.7 microM A23187 (a Ca2+ ionophore) increased intracellular Ca2+ concentration from mean (SEM) 14 (3) nM to 248 (30) nM and activated K+ channels in 21% of patches. It is concluded that the plasma membrane of HGT-1 cells possesses (a) specific 3H-omeprazole binding sites, which may reflect the omeprazole sensitive H+,K(+)-ATPase present in gastric parietal cells; and (b) Ca(2+)-activated K+ channels, which may be located in the basolateral membrane of human gastric parietal cells and play a part in acid secretion triggered by Ca(2+)-mediated secretory agonists. PMID- 8244098 TI - Systemic and mucosal humoral responses to Helicobacter pylori in gastric cancer. AB - The systemic IgG response to Helicobacter pylori was examined in 70 patients with gastric cancer. H pylori IgG antibodies were assayed by enzyme linked immunosorbent assay (ELISA), and serological recognition of H pylori antigens was characterised by western blotting. A percentage of 78.5 were seropositive by ELISA. Two of five patients under age 50 were seronegative. Positivity was unrelated to age, sex, tumour type, or site. Ninety one per cent of ELISA positive cancer patients recognised the H pylori cytotoxin associated 120 kilodalton (kD) protein, significantly more than a control group of 47 ELISA positive patients with non-ulcer dyspepsia (72%). Four of 15 ELISA negative cancer patients also showed recognition of this protein in western blots. Mucosal IgA responses to H pylori were examined by immunoblotting supernatants of in vitro cultured resected antral mucosa in an overlapping group of 19 gastric cancer patients. Eighteen had a positive response, including 10 of 11 negative for H pylori by biopsy urease testing. The systemic and local immunoblotting results show that the high seroprevalence of H pylori antibodies detected by ELISA is nevertheless an underestimate of past infection. Dyspepsia screening policies based solely on H pylori ELISA would miss some young patients with gastric cancer. Further study of the relation of the H pylori cytotoxin to gastric precancerous lesions is warranted. PMID- 8244099 TI - Acute perforated duodenal ulcer is not associated with Helicobacter pylori infection. AB - Most patients with chronic duodenal ulcer disease have Helicobacter pylori infection and eradicating it considerably reduces the relapse rate. The prevalence of H pylori in 80 patients (mean age = 52 years, range 17-85) presenting with acute perforated duodenal ulcer was examined and compared with age and sex matched hospital control patients. H pylori state was assessed by serum anti-H pylori IgG (Helico-G kit, Porton) using a titre of 18 or less as negative with a specificity of 89% and sensitivity of 88%. Only 47% of the perforated duodenal ulcer patients were positive for H pylori and this was similar to the value of 50% in the controls. In 51 of the perforated duodenal ulcer patients 14C-urea breath tests were also performed 4-10 weeks after surgery and this confirmed that only 49% were positive for H pylori. None of these patients had received perioperative drugs that might have eradicated the infection. The H pylori positive and H pylori negative perforated duodenal ulcer patients were similar with respect to age (53, 51), smoking (84%, 83%), and consumption of more than 15 units of alcohol per week (42%, 38%). Duodenal ulcer disease had been diagnosed before acute perforation in only 24% of those with H pylori and also 24% of those without the infection. Regular non-steroidal anti inflammatory drug (NSAID) use was common in both those with (44%) and without (45%) H pylori. In conclusion, the lack of association of acute perforated duodenal ulcer and H pylori infection suggests that perforated duodenal ulcer has a different pathogenesis from chronic duodenal ulcer disease, and that the first should not be regarded simply as a complication of the second. PMID- 8244100 TI - Helicobacter pylori: comparison of DNA fingerprints provides evidence for intrafamilial infection. AB - Although a high prevalence of antibodies to Helicobacter pylori has been documented within families, culture and DNA typing of strains from infected children and their parents has not been evaluated. This study aimed to analyse H pylori infection within family groups. Endoscopy, gastric biopsy, and H pylori culture were performed on all eight parents of four children who presented with dyspepsia and who had a positive H pylori culture. All biopsy specimens were cultured on Columbia based blood agar under microaerophilic conditions for four days. The DNA from each strain was extracted and electrophoretic patterns were compared after digestion with restriction endonucleases Hae III or Hind III. Ribotyping using a biotinylated cDNA probe prepared from 16S and 23S rRNA of H pylori NCTC 11638 was also used. Seven of the parents were positive for H pylori on urease testing, histology, and on culture. DNA typing showed the same or a similar strain to be present in at least two family members in three of the four family groups. In family 1, the mother, father, and child all had an identical strain; in family 2, father and son had a similar related strain; father and mother had the same strain in family 3; and all strains were unique in family 4. These data provide evidence for either intrafamilial cross infection or a common source of infection within family groups. PMID- 8244101 TI - Distribution of macrophages and granulocytes expressing L1 protein (calprotectin) in human Peyer's patches compared with normal ileal lamina propria and mesenteric lymph nodes. AB - Antibodies to the cytosolic leucocyte L1 protein (or calprotectin) were examined for reactivity with macrophages, neutrophils, and eosinophils identified by paired immunofluorescence staining in sections of normal human ileal mucosa, including Peyer's patches. Macrophages were recognised by expression of the myelomonocytic antigen CD68 (monoclonal antibody KP1). Neutrophilic granulocytes were identified by their content of neutrophil elastase, and eosinophilic granulocytes by monoclonal antibody EG2. Virtually all CD68+ macrophages in normal lamina propria and Peyer's patches were L1- and the same was true for most extravasated macrophages in normal peripheral lymph nodes. Some mesenteric lymph nodes, however, and all peripheral lymph nodes with overt pathological processes (malignant lymphoma) contained many CD68+L1+ macrophages. Numerous L1+ cells were also localised to the crypt region and to some extent beneath the villous epithelium in normal lamina propria, but they were mainly identified as EG2+ eosinophils. Such cells were remarkably scarce or absent beneath the follicle associated epithelium in the dome region of Peyer's patches, where CD68+L1- macrophages were abundant. Also subepithelial and interfollicular CD68- interdigitating dendritic cells in Peyer's patches (recognised by antibody to S 100 protein) were usually unreactive with L1 antibody. The L1 protein shows a broad spectrum of antimicrobial activities in vitro, and its putative antiproliferative properties are interesting in relation to the immunosuppression postulated to take place in lamina propria. The virtual absence of L1 producing cells beneath the follicle associated epithelium in Peyer's patches may support the immunostimulatory function of these macrophage rich structures, which are held to be crucial for induction of specific mucosal immunity. PMID- 8244102 TI - Technetium-99m hexamethyl propylene amine oxime granulocyte scintigraphy in Crohn's disease: diagnostic and clinical relevance. AB - Scintigraphy with autologous granulocytes labelled by technetium-99m hexamethyl, propylene amine oxime (99mTc-HMPAO) was performed in 103 Crohn's disease patients and 52 healthy controls. In 31 patients endoscopic and histologic activity was compared with scan activity index. In the 98 patients with a positive scan, the extent of Crohn's disease, assessed by scintigraphy, was compared with that evaluated by small bowel x ray or colonoscopy with biopsies. In 48 patients, Crohn's disease activity index, activity index, simple index, erythrocyte sedimentation rate, C-reactive protein were correlated with the scan results. In 16 patients the five parameters and scan were repeated after treatment with methyl-prednisolone (10 cases), enteral nutrition (3), and 5-acetylsalicylic acid (3). The results showed that 99mTc-HMPAO granulocyte scan had a 95% sensitivity and 100% specificity to detect active inflammation; it correctly showed an abscess or a fistula in all the 24 cases found. The correlation between histological inflammatory activity and scan activity index was highly significant (r = 0.85; p < 0.01), less significant (r = 0.65; p < 0.01) between endoscopy and scan activity index. The evaluation for the extent of Crohn's disease by scan was completely correct in the small bowel (100%) and 93% correct in the large bowel. No correlation was seen between the three clinical activity parameters and scanning; in more than 80% of the cases in remission on the basis of a clinical or laboratory index, scintigraphy remained positive. Medical treatment was effective on the clinical indices but not on the active inflammation in the ileum, whereas it led to a negative scan in 5/11 cases in the large intestine. Scintigraphy with 99mTc-HMPAO granulocyte plays an important part in Crohn's disease for the diagnosis of complications, for activity and assessment of the extent, and for the treatment results evaluation. PMID- 8244103 TI - Abnormal plasma polyunsaturated fatty acid pattern in non-active inflammatory bowel disease. AB - An abnormal plasma polyunsaturated fatty acid pattern (PUFA) (increased n3 and decreased n6 PUFA) has been reported in active inflammatory bowel disease (IBD). The possibility of a primary defect in the PUFA metabolism in IBD was hypothesised. The aim of this study was to assess plasma PUFA pattern in inactive inflammatory bowel disease and to ascertain whether patients who had had a colectomy and who were suffering from ulcerative colitis have a similar PUFA pattern than those patients with non-active ulcerative colitis and who had not had a colectomy. Plasma fatty acids were analysed by semi-capillary column gas liquid chromatography in three groups of patients with inactive IBD (24 patients with inactive ulcerative colitis who had not had a colectomy, 15 patients with ulcerative colitis who had had a colectomy, and 27 patients with Crohn's disease). Plasma concentration and percentage of C22:6n3 and unsaturation index were significantly higher in patients with inactive ulcerative colitis without a colectomy and the Crohn's disease group (p < 0.0001) than in controls. Plasma concentration and percentage of C22:6n3 and the unsaturation index remained significantly higher, in both the operated and non-operated ulcerative colitis patients when compared with controls (p < 0.0001). These results suggest that in inactive IBD, an increased PUFA biosynthesis might be the cause of the high values of n3 compounds. These findings although seen in active disease, are more noticeable in remission because of the lack of artefactual factors (malnutrition, steroids, inflammation). In addition, persistence of high values in both groups of ulcerative colitis patients--that is, those who had had a colectomy and those who had not suggests the existence of a primary abnormality in the PUFA metabolism in IBD. PMID- 8244104 TI - Diffuse jejunoileitis of Crohn's disease. AB - Diffuse jejunoileitis is an uncommon, but important manifestation of Crohn's disease because of the associated high morbidity and challenges in medical management. Despite this there have been no studies of prognosis or management of diffuse jejunoileitis for nearly 20 years. This study analysed the outcome among 34 patients (20 women and 14 men) with diffuse jejunoileitis diagnosed between 1960 and 1991 including clinical features, medical and surgical management, death rates, current status, and prognosis. Diffuse jejunoileitis presents in younger patients (mean age at diagnosis 26.4 years) than those with distal ileal Crohn's disease (mean age at diagnosis 33.3 years). Nearly all presented with clear cut abdominal symptoms including a combination of colicky abdominal pain (91%), weight loss (62%), and diarrhoea (53%). Most patients had severe symptoms reflected by the fact that 77% had been treated with corticosteroids for periods of more than six months at some stage during their follow up. The mean follow up from diagnosis was 16 years. Twenty eight patients (82%) had at least one operation for diffuse jejunoileitis and two thirds of the patients (n = 21) required two or more operations. The frequency of surgical intervention was particularly high in the younger patients (r = 0.71, p < 0.001). The proportion of patients requiring surgery was highest in the first year after diagnosis. The annual operative rate was 15% for the first 10 years and then it fell to 5.2% in years 11-15, and 2.6% in years 16-20. The data suggest that the disease does burn itself out over time. The increasing use of strictureplasty for short strictures and the minimal use of resection has eliminated problems associated with the short small bowel syndrome. The longterm prognosis of this patients is good. Only two patients died (one of perforation of the jejunum and another of an unrelated bronchogenic carcinoma). After a mean interval from diagnosis of 16 year, 24 of 32 living patients are well and symptom free. Only eight have abdominal symptoms of whom three are receiving corticosteroid treatment and one azathioprine. The combination of anti-inflammatory drugs with the relief of recurrent obstructive symptoms by strictureplasty can together produce a good longterm prognosis in most patients with diffuse jejunoileitis. PMID- 8244105 TI - Carriage of adhesive Escherichia coli after restorative proctocolectomy and pouch anal anastomosis: relation with functional outcome and inflammation. AB - Restorative proctocolectomy with pelvic ileal reservoir is a well accepted option for the surgical treatment of ulcerative colitis. Acute pouchitis is a common complication and resembles acute ulcerative colitis. Patients with ulcerative colitis carry Escherichia coli that adhere to epithelial cells and thus this study examined whether acute pouchitis is associated with the carriage of adhesive E coli. E coli isolated from stool samples from 24 patients (median age 34 years, range 16-64; 13 men, 11 women) who had had restorative proctocolectomy with pelvic ileal reservoir were examined by means of the buccal epithelial cell adhesion assay. Patients were studied at a median of 12 months (range 7-21) after operation. Eight of 24 patients had acute pouchitis at the time of study. Adhesive E coli were detected in nine of 24 patients with a pelvic ileal reservoir compared with none of 12 controls (p < 0.05). The buccal epithelial cell adhesion index was inversely related to the degree of acute pouchitis (rs = 0.46, p = 0.024) and to the functional outcome (rs = -0.49, p = 0.022). Carriage of adhesive E coli was not related to the design of the reservoir. By contrast with ulcerative colitis, acute pouchitis is not associated with the carriage of adhesive E coli. PMID- 8244106 TI - Outcome of restorative proctocolectomy when the diagnosis is suggestive of Crohn's disease. AB - Twenty of 81 patients treated by restorative proctocolectomy for presumed ulcerative colitis had some features of Crohn's disease: 10 were classified as definite Crohn's disease and 10 as indeterminate colitis. These pathological features were first apparent during synchronous colectomy and pouch construction in 10 of 11 cases. In the remainder, histological features of possible Crohn's disease were first identified during rectal excision (n = 6), preliminary subtotal colectomy (n = 2), and after pouch excision (= 2). Complications were marginally more common in patients with features of possible Crohn's disease: pelvic sepsis 30% (Crohn's disease 30%, indeterminate colitis 30%) v 20%, fistulas 45% (Crohn's disease 30%, indeterminate colitis 60%) v 16%; ileal stenosis 40% (Crohn's disease 40%, indeterminate colitis 40%) v 21%, pouchitis 50% (Crohn's disease 50%, indeterminate colitis 50%) v 26%, and small bowel obstruction 25% (Crohn's disease 30%, indeterminate colitis 30%) v 13%. Pouch excision or a persistent proximal stoma has been necessary in six patients with possible Crohn's disease (30%) (Crohn's disease 3 cases 30%, indeterminate colitis 3 cases 30%) compared with nine (15%) of the remainder. Median hospital stay, however, was the same and stool frequency in those with a functioning pouch were comparable. These results show that there is a higher complication rate if there are features of Crohn's disease but that the medium term functional results are acceptable if the pouch can be retained. PMID- 8244107 TI - Non-penetrance and late appearance of polyps in families with familial adenomatous polyposis. AB - One case of non-penetrance of the familial adenomatous polyposis (FAP) gene at 59 years of age and late onset of polyps on endoscopy and biopsy in this and two other families is described. Screening protocols should include dental screening as well as indirect ophthalmoscopy and endoscopy to detect minimal manifestations of the gene. In the absence of a specific DNA predictive test, bowel screening should continue well beyond 30 years of age. PMID- 8244108 TI - Increased risk of thyroid and pancreatic carcinoma in familial adenomatous polyposis. AB - Familial adenomatous polyposis has been associated with several extraintestinal cancers, but the relative and absolute risks of these malignancies have not been determined. Extraintestinal cancers reportedly associated with adenomatous polyposis (thyroid gland, adrenal gland, pancreas, and biliary tract) were identified in polyposis patients and their at risk relatives in The Johns Hopkins Registry. The incidence rates of identified tumours were then compared with the general population through person year analysis with adjustment for population. For comparison, the incidence rates of the two most common cancers not associated with polyposis (breast cancer in women and lung cancer) were also calculated. There was an increased relative risk of thyroid cancer (relative risk 7.6; 95% confidence limits (CL) 2.5-17.7) and pancreatic adenocarcinoma (relative risk 4.46; 95% CL 1.2-11.4) in polyposis patients and at risk relatives. The absolute risk was 26.8 and 21.4 cases/100,000 person years, respectively. No cases of adrenal or biliary cancer were found in this cohort. There was no increased relative risk of lung cancer (95% CL 0.04-1.4) or breast cancer (95% CL 0.04-1.4) over the general population. The relative risks of thyroid and pancreatic cancer are increased in familial adenomatous polyposis, but the absolute lifetime risk is low. Screening for pancreatic cancer may not be worthwhile with currently available methods, but careful physical examination of the thyroid gland is warranted along with consideration for ultrasonography. PMID- 8244109 TI - Bowel dysfunction in young women with urinary retention. AB - Urinary retention in young women is rare, but a syndrome has recently been described in which urinary retention is associated with abnormal periurethral electromyography. The incidence of bowel dysfunction in these women was investigated, in an attempt to determine whether this might be a more widespread disorder. Of 12 patients with this urological abnormality, eight were constipated. No consistent anorectal abnormality was identified on anorectal physiological testing. The electromyographic abnormality was not seen in the external anal sphincter. One patient had an abnormality of the internal anal sphincter smooth muscle, while another had a generalised disorder of the gastrointestinal tract and urinary bladder resembling a visceral myopathy. Bowel symptoms are common in this group of women with urinary retention, but abnormalities of bowel function are not specific. A common mechanism for bladder and bowel symptoms remains a possibility. PMID- 8244110 TI - Constipation in early childhood: patient characteristics, treatment, and longterm follow up. AB - Little is known about chronic constipation in infants, toddlers, and preschool children and longterm outcome after treatment. The symptoms of 174 children < or = 4 years of age, who were evaluated for chronic constipation, are reported in this study together with the long-term outcome in 90 of them. Initial symptoms were infrequent bowel movements in 58%, painful bowel movements in 77% often with screaming, and severe stool withholding manoeuvres in 97%. The treatment of chronic idiopathic constipation consisted of education, faecal disimpaction, prevention of future impaction, and promotion of regular bowel habits with dietary fibre and milk of magnesia, and finally toilet training of the preschool child. Longterm outcome could be evaluated in 90 patients (52%) (mean (SD) 6.9 (2.7)) years after initial evaluation. Fifty seven children (63%) had recovered, defined as no soiling with > or = 3 bowel movements per week, while not receiving treatment. The recovery rate of children < or = 2 years of age was significantly higher than in children > 2 to 4 years of age. Thirty three children (37%) had not recovered. Constipation recurred as soon as laxatives were discontinued in 31 (94%) of them. Laxatives were still used by 33% of the children who had not recovered, 39% had < 3 bowel movements per week, 48% had faecal soiling, 45% had stool withholding, 27% complained of abdominal pain, 73% passed large stools, and 45% still on occasions clogged the toilet with their large stools. Symptoms of chronic constipation persisted in one third of our patients, 3-12 years after initial evaluation and treatment. Children who had not recovered deserve continued follow up, to reinforce and adjust treatment and to prevent faecal soiling. PMID- 8244111 TI - Intestinal pseudo-obstruction in patients with amyloidosis: clinicopathologic differences between chemical types of amyloid protein. AB - A clinicopathologic study was made of 16 patients with amyloidosis and with clinical signs of intestinal pseudo-obstruction. amyloid deposits in the small intestine were proved in all cases by endoscopic or intra-operative biopsies, and immunohistochemical study identified the chemical types of amyloid protein: amyloid A protein (AA) in 13 cases, light chain protein (AL) in two, and beta 2 microglobulin (AH) in one. Clinically, an acute self limiting obstructive condition was evident in 13 cases with AA, and 12 of them returned to normal bowel function after receiving total parenteral nutrition. Two cases with AL and one with AH presented chronic, intermittent, obstructive symptoms, and medical treatment, including total parenteral nutrition, was ineffective with no recovery of intestinal propulsion. Pathological examination of the necropsy specimens in seven cases showed considerable differences in the preferential sites of gastrointestinal deposits between the chemical types of amyloid; extensive infiltration and replacement of the muscularis propria by amyloid deposits throughout the gastrointestinal tract, especially the small intestine, were found in the AL and the AH cases, while amyloid deposits in the myenteric plexus without appreciable muscle infiltration were shown in the AA cases. These results show that intestinal pseudo-obstruction in patients with amyloidosis is caused by either myopathy or neuropathy, and that chemical types of amyloid may determine which of the two factors has the dominant affect on the bowel function. PMID- 8244112 TI - Dieulafoy's disease: endoscopic treatment and follow up. AB - The findings from 480 patients who had emergency endoscopy for acute upper gastrointestinal bleeding of non-variceal origin at our institution were analysed. Twenty eight patients (5.8%) had a Dieulafoy lesion. In 27 patients (96.4%) bleeding could be successfully managed by injection of norepinephrine and polidocanol, in repeated sessions if needed. Two patients had to be treated surgically: one because of uncontrollable bleeding from the Dieulafoy lesion and one despite endoscopic control of the bleeding Dieulafoy lesion because of a concomitant bleeding from an anastomosal ulcer after gastric resection. Three patients died during hospital stay from causes unrelated to bleeding from Dieulafoy lesion. Out of the 25 patients discharged from the hospital 21 treated by endoscopy and two treated with surgery were followed up for a mean of 28.3 and 22.5 months, respectively. Twenty endoscopically treated patients (95%) had no recurrence of Dieulafoy's bleeding. One patient experienced severe rebleeding from the original site after a transient endoscopy confirmed complete disappearance. He had emergency operation without a further attempt to control bleeding by endoscopy. It is concluded that bleeding from Dieulafoy's disease can be successfully managed by endoscopic injection treatment. The longterm outcome is favourable. PMID- 8244113 TI - Transrectal ultrasound study of the pathogenesis of solitary rectal ulcer syndrome. AB - Transrectal ultrasonography is of clinical value in anorectal carcinoma and in inflammatory diseases of the anorectum. In this study a rigid linear endorectal probe was used to examine 15 patients with endoscopically and biopsy proved diagnosis of solitary rectal ulcer syndrome. In 13 of the 15 patients the rectal wall was thicker (mean (SEM) 5.7 (0.4) mm; normal values: 2.8 (0.1) mm) near the rectal ulcer. In all these cases the muscularis propria layer exceeded the maximum normal diameter of 2 mm. In nine of the 15 patients the normal rectal wall echo-structure, with five distinct layers, was disturbed and there was fading of the borders between the mucosa and the muscularis propria. Poor relaxation of the puborectalis muscle during straining was seen on ultrasound in 11 patients, as was intussusception of the rectal wall. The obvious enlargement of the muscularis propria points to a chronic mechanical load on the rectal wall. The ulcerative lesions are formed in this area of overloaded rectal wall. The direct visualisation of the puborectalis muscle during dynamic transrectal ultrasonography indicates that the fact that it does not relax is an important element in the pathogenesis of solitary rectal ulcer syndrome. PMID- 8244114 TI - Iron deficiency anaemia--how far to investigate? AB - Since 1980 investigations, by this group, of patients with iron deficiency anaemia and no obvious cause, have been limited to upper gastrointestinal endoscopy, with small bowel biopsy, and barium enema. This study attempted to follow up 93 consecutive patients whose anaemia remained obscure after these investigations to determine the outcome and assess the safety of this limited approach. In 1991/92 questionnaires were sent to the general practitioners. Eighty three completed questionnaires were received. Ten patients had died all unrelated to the iron deficiency anaemia. The mean follow up of the 73 living patients was six years (range 4-12). Sixty five (89%) had a normal haemoglobin concentration and only 15 were still taking oral iron. Possible causes for the anaemia were found in 17-non-steroidal anti-inflammatory drug use in 10, menstruation in two, gastrectomy in three, poor nutrition in two. No other cause emerged. It is concluded that this limited investigative approach is safe. PMID- 8244115 TI - Audit of endoscopic surveillance biopsy specimens in HIV positive patients with gastrointestinal symptoms. AB - An audit of upper gastrointestinal endoscopy in HIV infected patients with gastrointestinal symptoms assessed the frequency of disease detected by endoscopy and routine laboratory analysis of surveillance biopsy specimens. Sixty nine consecutive endoscopies were performed in 59 HIV infected patients. Endoscopic biopsy specimens were taken from the lower oesophagus, gastric antrum, and third part of the duodenum for virology, histopathology, parasitology, bacteriology, and mycobacterial culture. Endoscopic appearances detected disease in 25/59 (42.4%) patients (oesophageal candida, 14; oesophageal ulcer, 3; Kaposi's sarcoma, 4; others, 4), but only 4/43 (9.3%) specimens showed evidence of disease in the absence of endoscopic abnormality. Virology for cytomegalovirus (detection of early antigenic fluorescent foci and culture) was positive in 6/59 (10.2%) patients, but parasitology and mycobacterial culture were negative in all cases. Histopathology was abnormal in 11/52 (21%) oesophageal biopsy specimens, 13/47 (28%) gastric biopsy specimens, and 4/65 (6%) duodenal biopsy specimens. Abnormal findings were found predominantly in those with advanced HIV disease (CDC Stage IV) (21/33 patients (64%)) compared with those with early HIV disease (CDC Stage II) (5/26 (19%)). In conclusion, upper gastrointestinal endoscopy detects macroscopic disease in AIDS patients and those with low CD4 counts, but routine surveillance biopsy specimens of apparently normal bowel in early HIV disease (or where CD4 counts are greater than 0.2 x 10(9)/1) are of little value. PMID- 8244116 TI - Human gastric alcohol dehydrogenase activity: effect of age, sex, and alcoholism. AB - As various isoenzymes of gastric alcohol dehydrogenase exist and as the effect of sex and age on these enzymes is unknown, this study measured the activity of gastric alcohol dehydrogenase at high and low ethanol concentrations in endoscopic biopsy specimens from a total of 290 patients of various ages and from 10 patients with chronic alcoholism. Gastric alcohol dehydrogenase was also detected by immunohistological tests in biopsy specimens from 40 patients by the use of a polyclonal rabbit antibody against class I alcohol dehydrogenase. A significant correlation was found between the immunohistological reaction assessed by the intensity of the colour reaction in the biopsy specimen and the activity of alcohol dehydrogenase measured at 580 mM ethanol. While alcohol dehydrogenase activity measured at 16 mM ethanol was not significantly affected by age and sex, both factors influenced alcohol dehydrogenase activity measured at 580 mM ethanol. Young women below 50 years of age had significantly lower alcohol dehydrogenase activities in the gastric corpus and antrum when compared with age matched controls (SEM) (6.4 (0.7) v 8.8 (0.6) nmol/min/mg protein; p < 0.001 and 6.0 (1.3) v 9.5 (1.3) nmol/min/mg protein; p < 0.001). Over 50 years of age this sex difference was no longer detectable, as high Km gastric alcohol dehydrogenase activity decreases with age only in men and not in women. In addition, extremely low alcohol dehydrogenase activities have been found in gastric biopsy specimens from young male alcoholics (2.2 (0.5) nmol/min/mg protein), which returned to normal after two to three weeks of abstinence. The activity of alcohol dehydrogenase in the human stomach measured at 580 mM ethanol is decreased in young women, in elderly men, and in the subject with alcoholism. This decrease in alcohol dehydrogenase activity may contribute to the reduced first pass metabolism of ethanol associated with raised ethanol blood concentrations seen in these people. PMID- 8244117 TI - Reversal of carbon tetrachloride induced changes in microviscosity and lipid composition of liver plasma membrane by colchicine in rats. AB - Colchicine is beneficial in the treatment of cirrhotic patients, it prevents changes in plasma membrane bound enzymes induced by CCl4 intoxication. In this study, lipid composition and microviscosity were measured in liver plasma membranes isolated from rats given CCl4. Microviscosity values increased in rats given CCl4 for six weeks but fell considerably in those given CCl4 for 10 weeks. Both these changes were absent when colchicine was given with CCl4. The cholesterol/phospholipid molar ratios and lipid peroxide values increased but plasma membrane phospholipids, the length of fatty acyl chains, and the unsaturation index fell significantly after CCl4 intoxication. Colchicine treatment also prevented these changes. Changes in the lipid composition of liver plasma membranes were significantly correlated with lipid peroxidation. Colchicine prevents changes in the physicochemical properties of liver plasma membranes induced by longterm CCl4 treatment, probably by blocking peroxidation of unsaturated fatty acids. PMID- 8244118 TI - Correlation between gall bladder fasting volume and postprandial emptying in patients with gall stones and healthy controls. AB - To evaluate whether the extent of postprandial gall bladder emptying is correlated with gall bladder fasting volume, gall bladder motility was studied in 56 patients with cholesterol gall stone and 19 control patients. Gall bladder volumes were determined sonographically, while cholecystokinin plasma values were measured radioimmunologically. Twenty three per cent of gall stone patients were classified as pathological contractors (residual fraction > mean +2SD of controls) and 77% as normal contractors. Normal but not pathological contractor patients exhibited larger gall bladder fasting volumes (mean (SEM)) (24.7 (1.7) ml) than controls (15.3 (1.2) ml, p < 0.001). In normal contractor patients and controls fasting volume was closely related with ejection volume (r = 0.97, p < 0.001) and residual volume (r = 0.80, p < 0.001). Although ejection volume was enlarged in normal contractor patients it did not compensate the increase in fasting volume. Thus, residual volumes were considerably increased not only in pathological contractors (12.7 (2.5) ml, p < 0.001) but also in normal contractor patients (7.0 (0.5) v 4.6 (0.6) ml, p < 0.001). Postprandial cholecystokinin secretion did not differ between patients and controls. It is concluded, that in normal contractor patients gall bladder fasting volume is closely correlated with ejection and residual volume. Thus, fasting volume may be an essential factor affecting postprandial gall bladder emptying. Large fasting volumes in cholesterol gall stone disease could thereby contribute to bile retention, which facilitates gall stone growth. PMID- 8244119 TI - Outcome after cholecystectomy for symptomatic gall stone disease and effect of surgical access: laparoscopic v open approach. AB - The pre and postoperative symptoms and outcome after surgery in patients with symptomatic gall stone disease were evaluated by a detailed self administered postal questionnaire. The survey was conducted in two groups: 80 patients treated by laparoscopic cholecystectomy and an age matched cohort of patients who had conventional open cholecystectomy. The overall response rate on which the data were calculated was 76%. Symptomatic benefit ratios accruing from the surgical removal of the gall bladder were calculated. The symptoms that were relieved by cholecystectomy were nausea (0.98), vomiting (0.91), colicky abdominal pain (0.81), and backpain (0.76). Flatulence, fat intolerance, and nagging abdominal pain were unaffected as shown by a benefit ratio of 0.5 or less. Relief of heartburn (39/49) outweighed the de novo development of this symptom after cholecystectomy (7/49), resulting in a benefit ratio of 0.65. Postcholecystectomy diarrhoea occurred in 21/118 patients (18%): 10 after open cholecystectomy and 11 after laparoscopic cholecystectomy. The type of surgical access did not influence the symptomatic outcome but had a significant bearing on the time to return to work or full activity after surgery (laparoscopic cholecystectomy two weeks, open cholecystectomy eight weeks, p = 0.00001). In the elderly age group (> 60 years), significantly more patients (29/30) regained full activity after laparoscopic cholecystectomy when compared with the open cholecystectomy group (16/22), p = 0.001. The patient appreciation of a satisfactory cosmetic result was 72% in the open group compared with 100% of patients who were treated by laparoscopic cholecystectomy (p = 0.0017). Despite the persistence or de novo occurrence of symptoms, 111/117 patients (95%) considered that they had obtained overall symptomatic improvement by their surgical treatment and 110/118 (93%) were pleased with the end result regardless of the access used. PMID- 8244120 TI - Expression of major histocompatibility antigens in human chronic pancreatitis. AB - T-lymphocytic infiltration of the exocrine pancreas and liver in patients with chronic pancreatitis has suggested that cell mediated immune mechanisms may play a part in the pathogenesis of this disease. As expression of major histocompatibility (MHC) antigens is a prerequisite for organ specific autoimmunity, the expression of HLA class I (beta 2-microglobulin) and class II (HLA-DR) determinants have been analysed, together with the presence of T lymphocytes, in 93 patients (64 men and 29 women, mean age 40.6 years) having an operation for chronic pancreatitis. Ethanol (63 patients), recurrent acute pancreatitis (12), congenital lesions (2), and unknown (16) were suggested to be the causes of the disease. Immunohistochemical staining of formalin fixed and paraffin wax embedded tissue sections used conventional immunohistochemical techniques with specific anti-serum samples. No MHC expression was identified in 10 histologically normal pancreatic control specimens or in four cases of chronic pancreatitis secondary to obstruction by neuroendocrine tumours within the head of the pancreas. beta 2-microglobulin expression by pancreatic exocrine epithelial cells was seen in 76 chronic pancreatitis specimens (82%) while HLA-DR was present in 61 (66%). Simultaneous expression of both class I and II determinants was seen in 53 (57%) of cases. MHC determinant expression was not found in 10 cases (11%) of chronic pancreatitis. In the positive specimens, expression was confined to ductal and ductular (interlobular and intralobular) epithelium with no staining of acinar cells. Staining was not related to the suspected cause of the disease or age. T-lymphocytes were more prominent in chronic pancreatitis mean (SEM) (131 (15) cells per high powered field) than controls (5 (1), p < 0.01). Aberrant MHC expression by exocrine pancreatic epithelial cells occurring in the presence of an appreciable T-cell infiltration confirmed that the appropriate cellular conditions were present for cell mediated cytotoxicity to contribute to the pathogenesis of chronic pancreatitis. PMID- 8244121 TI - Improved symptom relief and duodenal ulcer healing with lansoprazole, a new proton pump inhibitor, compared with ranitidine. AB - The purpose of this study was to compare duodenal ulcer healing, symptom relief, and safety of lansoprazole (a new proton pump inhibitor) given at doses of 30 mg and 60 mg, in the morning with ranitidine 300 mg at bedtime. Two hundred and eighty nine patients were enrolled over a 20 month period in a double blind randomised parallel group comparative study set in outpatient endoscopy units of six United Kingdom medical centres. Patients were randomised to receive lansoprazole 30 mg in the morning (n = 95), 60 mg in the morning (n = 96), or ranitidine 300 mg at bedtime (n = 98) for four weeks. Efficacy was assessed by gastroscopy at study entry and after two and four weeks of treatment. Symptom relief was monitored by patient diaries and physician review at two and four weeks. Both doses of lansoprazole resulted in significantly greater ulcer healing than ranitidine after two and four weeks. Respective healing rates on lansoprazole 30 mg, 60 mg, and ranitidine 300 mg were 78%, 80%, and 60% after two weeks and 93%, 97%, and 81% after four weeks. Patients on lansoprazole 30 mg (p = 0.002) and lansoprazole 60 mg (p = 0.026) also recorded greater relief of night time pain in the diary cards during the first seven days of treatment than those on ranitidine. Patients on lansoprazole 60 mg reported significantly better pain relief at their two week visit compared with those receiving ranitidine (p = 0.007). There were no differences between treatment groups in the occurrence or pattern of adverse drug reactions during the trial. It is concluded that for patients with duodenal ulcer, lansoprazole 30 mg or 60 mg is associated with faster ulcer healing and better symptom relief than ranitidine 300 mg at bedtime. There were no significant differences between lansoprazole 30 mg and 60 mg. These data indicate that lansoprazole should be used at a once daily dose of 30 mg for the treatment of duodenal ulcer. PMID- 8244122 TI - Pleural lymphoma in a patient presenting with malabsorption: an illustration of the clinicopathological behaviour in a case of enteropathy associated T cell lymphoma. AB - A case of enteropathy associated T cell lymphoma (EATCL) is described, which was diagnosed by biopsy of a lymphomatous pleural mass. Retrospective radiological review showed that this lesion had been present when an initial diagnosis of coeliac disease had been made 12 months previously and a detailed description of the natural history of the lymphoma during this period was thus available. The findings show that EATCL may behave in an indolent fashion and masquerade as coeliac disease, delaying the correct diagnosis. The relation of this disorder with coeliac disease and lymphocytic gastritis is discussed with reference to recent published works. PMID- 8244123 TI - Interleukin-6 in the early assessment of acute pancreatitis. PMID- 8244124 TI - Serological diagnosis of Helicobacter pylori. PMID- 8244125 TI - Pioneer in the gluten free diet: Willem-Karel Dicke 1905-1962, over 50 years of gluten free diet. PMID- 8244126 TI - Molecular biology. Setting the stage in colorectal cancer? PMID- 8244127 TI - Paediatric gastro-oesophageal reflux: prognostic indicators from pH monitoring. AB - The original diagnostic 24 hour pH monitoring data in 57 children with gastro oesophageal reflux (GOR) were retrospectively reviewed after a minimum of one year follow up. The tracings of children who responded to medical treatment were compared with those who failed to respond and required a fundoplication. Children with GOR secondary to oesophageal atresia/tracheo-oesophageal fistula and neurological conditions (n = 12) were analysed separately from those with primary GOR (n = 45). Children with primary GOR requiring a fundoplication (n = 9) had increased daytime reflux. The percentage time pH < 4 was the best discriminator (21% v 7%) with a threshold of 18% giving a 92% specificity and a 70% sensitivity. For children with secondary GOR the percentage time pH < 4 at night was significantly higher (29% v 3.7%) in those requiring a fundoplication (n = 5). A threshold of 18% gave an 80% specificity and an 86% sensitivity. These results show that both daytime and night time pH monitoring data can be of prognostic value in different subgroups of children with GOR. A percentage time pH < 4 of greater than 18% was a useful threshold to apply when evaluating the pH monitoring data. PMID- 8244128 TI - Non-invasive detection of gastro-oesophageal reflux using an ambulatory system. AB - The aim of this study was the separate measurement of reflux of food and acid into the oesophagus in 37 healthy, ambulant subjects. This was performed by radiolabelling the food and monitoring its reflux with a small directional gamma detector, which was placed externally over the oesophagus, and connected to an ambulatory data recorder. The pH was measured with a conventional oesophageal pH electrode. This method permitted the separate characterisation of acid and neutral (food) components of gastro-oesophageal reflux. The gastric emptying characteristics of the test meal were also monitored by gamma scintigraphy in a separate experiment. The oesophageal pH fell below 4 for 3.2 + 8.6/-2.3% (mean (SD)) of the recording time. Food reflux alone occurred for 17.8 + 53.2/-13.8% of the recording time. Simultaneous food and acid reflux occurred for only 0.95 + 5.2/-1.2% of the time. Not every reflux event detected by a fall in pH was seen as an increase in counts as a result of reflux of food, and vice versa. This poor correlation of food and acid reflux implies incomplete mixing of food and acid in the stomach, and further shows the inadequacy of reflux diagnosis methods that depend on pH detection alone. PMID- 8244129 TI - Acid clearance and oesophageal sensitivity in patients with progressive systemic sclerosis. AB - This study examined the hypothesis that impaired oesophageal peristalsis was associated with delayed oesophageal clearance of acid in patients with progressive systemic sclerosis (PSS), some of whom are thought to have impaired oesophageal sensitivity to acid. Sixteen patients with PSS had: (a) oesophageal manometry and endoscopy; (b) acid perfusion of the oesophagus with simultaneous measurement of intraoesophageal pH during perfusion and for the next 10 minutes; (c) 22 hour monitoring of intraoesophageal pH; and (d) an evaluation of reflux symptoms during and after perfusion and during overnight pH monitoring. By oesophageal manometry, eight patients had normal peristalsis and eight patients had impaired peristalsis. Oesophageal endoscopy was unremarkable in patients with normal peristalsis, whereas all patients with impaired peristalsis had oesophagitis. The time needed to clear the oesophagus of perfused acid was shorter (p < 0.01) in patients with normal peristalsis and acid clearance time was significantly correlated (p < 0.01) with acid exposure time during overnight pH monitoring. During and after oesophageal perfusion, the nature, duration, and severity of symptoms did not differ between the groups, but overnight symptoms lasted longer (p < 0.05) in patients with impaired peristalsis. It is concluded that in PSS: (1) Impaired oesophageal motility delayed the clearance of acid and increased the exposure time to acid. (2) Acid clearance time is a useful parameter of impaired oesophageal motor function. The assessment of acid clearance time can be used as an alternative to overnight pH monitoring, to assess the impairment of oesophageal acid clearance. (3) Oesophageal sensitivity to acid was preserved in patients with impaired peristalsis and oesophagitis. (4) Reflux symptoms lasted longer in patients with prolonged oesophageal acid exposure but were still reported for a small fraction of the total acid exposure time. Thus, reflux symptoms reflect poorly prolonged exposure of the oesophagus to acid and are not a reliable guide to acid injury of the oesophagus in PSS. PMID- 8244130 TI - Effects of supplemental oxygen on cardiac rhythm during upper gastrointestinal endoscopy: a randomised controlled double blind trial. AB - To investigate the effects of supplemental oxygen on cardiac rhythm during gastroscopy, 103 patients aged over 60 were randomised to receive either supplemental oxygen or air at 2 litres/minute during the procedure. Pulse rate, blood pressure, oxygen saturation, and a Holter cardiac trace were monitored before, during, and for one hour after the gastroscopy. A wide range of electrocardiographic abnormalities were recorded in both oxygen and air groups, of which ventricular and supraventricular ectopic beats were the most common. There were no significant differences in the rate of occurrence of any clinically important cardiac abnormality either between the oxygen and air groups or between the three monitored periods before, during, and after gastroscopy. There were significantly fewer patients, however, with supraventricular extra systoles when oxygen was given during gastroscopy (p < 0.05). Although supplemental oxygen during gastroscopy significantly improved oxygen saturation (p < 0.001; 95% confidence intervals for the difference between the means: 2.9 to 4.7), there was no correlation between oxygen saturation and any electrocardiographic changes. It is concluded that electrocardiographic abnormalities are common in patients over 60, but this study found no evidence that they are induced by gastroscopy. Supplemental oxygen increases oxygen saturation but does not reduce the incidence of clinically important cardiac arrhythmias. PMID- 8244131 TI - Long term results of endoscopic dilatation for corrosive oesophageal strictures. AB - Although dilatation is the treatment of choice for most patients with benign oesophageal strictures, there is little information on its efficacy and safety in corrosive oesophageal strictures. Of 123 adults with benign oesophageal strictures treated by endoscopic dilatation, 52 (42.3%) had strictures after corrosive ingestion and 39 (31.7%) had peptic strictures. Treatment was considered adequate if the oesophageal lumen could be dilated to 15 mm and there was complete relief of dysphagia. If dysphagia recurred after adequate initial dilatation, the stricture was dilated again up to 15 mm. Initial dilatation was adequate in 93.6% of patients with corrosive strictures and this success rate was comparable with that of the peptic stricture group (100%, p > 0.05). Long term success after adequate initial dilatation was studied in 36 patients with corrosive strictures (mean follow up 32.36 (17.12) months, range 6-60) and 33 patients with peptic strictures (mean follow up 36.32 (17.9) months, range 6-60). The mean (SEM) number of symptomatic recurrences per patient month during the total follow up period in the corrosive group was significantly higher than that in the peptic group (0.27 (0.04) v 0.07 (0.02), p < 0.001). The recurrence rate in the corrosive group, however, decreased over time, and after 12 months it was significantly (p < 0.001) lower than the recurrence rate in the first six months. After 36 months, the difference in the recurrence rate in the two groups was not significant (p > 0.05). Only nine oesophageal perforations occurred during a total of 1373 dilatation treatments (procedure related incidence 0.66%), and eight of these were in the corrosive stricture group. These patients were managed conservatively and subsequently strictures were dilated adequately in all. Endoscopic dilatation is safe and effective for short and long term relief of dysphagia in patients with corrosive oesophageal strictures. PMID- 8244132 TI - Efficacy of the heater probe in peptic ulcer with a non-bleeding visible vessel. A controlled, randomised study. AB - A controlled, randomised study was performed to evaluate the efficacy of treatment with heater probe in the prevention of rebleeding from peptic ulcer with a non-bleeding visible vessel. One hundred and one patients were randomised into two groups: patients to be treated by heater probe (n = 51) and controls without active treatment (n = 50). In the heater probe group rebleeding occurred in five patients (10%) v 13 (26%) in the control group (p = 0.03), with a comparative risk of 0.38 in favour of the heater probe group. The difference in proportions of successful treatment for each group was 16.2% in favour of the heater probe (95% CI = 2 to 31%). Haemorrhage directly related to heater probe treatment occurred in four patients. In three of them bleeding was easily controlled by further heater probe pulses. There were no other complications and no death in the heater probe group. One patient in the control group died of pulmonary embolism. No significant differences in the length of stay in hospital, blood transfusions, surgical rates, or death were found; the design of the study, however, precluded an adequate assessment of these variables, because the heater probe was an optional rescue treatment when high surgical risk patients rebled. These results suggest that the heater probe is an effective and safe procedure in the prevention of recurrent haemorrhage in peptic ulcer with a non-bleeding visible vessel. PMID- 8244133 TI - Ultrastructural patterns of Helicobacter pylori. AB - Ultrastructural morphology of the bacterial bodies was studied in 40 Helicobacter pylori positive cases. Two bacterial patterns were identified, which were associated with different modes of contact with the epithelial cells and possibly with different stages of the natural history of the infection. PMID- 8244134 TI - Gastric metaplasia and Helicobacter pylori infection. AB - Duodenal and antral mucosal biopsy specimens were obtained from 139 patients with dyspeptic complaints to study the prevalence and extent of gastric metaplasia in the duodenal bulb in relation to Helicobacter pylori (H pylori) infection and duodenal ulcer disease. On logistic regression, the presence and extent of gastric metaplasia was not significantly associated with H pylori infection. The prevalence of gastric metaplasia, however, was found to be higher in patients with current or past evidence of duodenal ulcer disease in comparison with subjects with functional dyspepsia (p = 0.01). A follow up study on 22 patients before and at least one year after eradication of H pylori showed that the mean extent of gastric metaplasia did not change significantly after eradication and did not differ when compared with 21 patients with persisting infection. It is concluded that the unchanged gastric acid output after eradication of H pylori is a more important factor in the development of gastric metaplasia than the H pylori related inflammatory process. PMID- 8244135 TI - Dose dependent effects of protracted ingestion of small amounts of gliadin in coeliac disease children: a clinical and jejunal morphometric study. AB - This study aimed to investigate the effects of chronic ingestion of small amounts of gliadin on children with coeliac disease. A four week challenge was performed on 20 children who had been on a gluten free diet for mean (SD) 14 (3) months. They were given a daily dose of either 100 mg (group A, n = 10, mean age 4 (2) years) or 500 mg of gliadin (group B, mean age 5 (3) years). The effects of the gliadin were monitored by morphometric study of the jejunal mucosa, intestinal permeability test with cellobiose/mannitol, and serum antigliadin antibody test. After the challenge, group A patients showed a significant increase in the mean intraepithelial lymphocyte count (before challenge 11 (3), afterwards 19 (6)) and a decrease in the villous height/crypt depth ratio (beforehand 1.5 (0.1), afterwards 1.3 (0.2)), while the intestinal permeability test remained normal and the IgA-antigliadin antibody increased in four of 10 children. After the challenge group B showed more pronounced histological changes, an increase in the mean urinary cellobiose/mannitol % (beforehand 0.028 (0.020), afterwards 0.058 (0.028)), and IgA-antigliadin antibody positivity in six of eight subjects. The discriminant analysis function showed that the pretreatment group, group A after challenge, and group B after challenge were correctly classified in 90% of cases by functions based on the individual intraepithelial lymphocyte count and the villous height/crypt depth ratio. This study shows that chronic ingestion of small amounts of gluten causes dose-dependent damage to the small intestinal mucosa in children with coeliac disease. The predictive value of laboratory tests, such as the antigliadin antibody test and the intestinal permeability test seems to be lower in treated patients than in those with active coeliac disease. PMID- 8244136 TI - Effect of a satiating meal on the concentrations of procolipase propeptide in the serum and urine of normal and morbidly obese subjects. AB - The effect of a satiating meal on the serum and urinary concentrations of procolipase propeptide (Ala-Pro-Gly-Pro-Arg, APGPR) immunoreactivity, as measured by enzyme linked immunosorbent assay (ELISA) specific for free APGPR, has been studied in normal and morbidly obese human subjects. The normal subjects displayed a biphasic response with coordinate increases in both serum and urine APGPR immunoreactivity both occurring within the first two hours after the meal. In two of three of the morbidly obese subjects, this early rise in APGPR concentration in urine was not seen but was followed by a slow rise in urinary APGPR immunoreactivity at four to six hours. In both the normal and obese groups, the urinary immunoreactive signal was found to coelute with synthetic APGPR on gel chromatography. In rats, procolipase propeptide (Val-Pro-Asp-Pro-Arg, VPDPR) specifically inhibits fat intake early in the postprandial period when given peripherally or centrally. This study suggests that in humans APGPR reaches the circulation shortly after feeding and is excreted in the urine. These findings are consistent with the hypothesis that human procolipase propeptide may also act as a satiety signal. In addition the late appearance of the peptide in some of the morbidly obese patients could be associated with perturbation of appetite control in these subjects. PMID- 8244137 TI - Nerve involvement in fluid transport in the inflamed rat jejunum. AB - Net fluid transport was measured in denervated jejunal segments of rats infected with larvae of Nippostrongylus brasiliensis. On days 6-9 after nematode inoculation, when the jejunal segment exhibited macroscopic and microscopic signs of inflammation, net fluid absorption was noticeably attenuated compared with control, and in eight of 26 experiments a net fluid secretion was seen. To determine whether enteric nerves participated in the response, intravenous hexamethonium (10 mg/kg body weight) was given or lidocaine (1% solution) was placed on the serosa of the intestinal segment. Both drugs significantly reduced fluid secretion or increased fluid absorption. The effect was more pronounced the lower the rate of fluid absorption or the higher the rate of fluid secretion. The inflammatory response influenced intestinal fluid transport partly via activation of the enteric nervous system. It was estimated that 50-60% of the change in fluid transport caused by the parasite could be ascribed to activation of intramural nervous reflexes. The effect of hexamethonium indicates that a cholinergic synapse is present in the secretory nervous reflux activated by inflammation. Experiments were also performed on animals on days 11-14 after infection when the nematodes had been expelled from the animal. A large net fluid absorption was then recorded. PMID- 8244138 TI - Neoterminal ileal blood flow after ileocolonic resection for Crohn's disease. AB - Endoscopic laser Doppler flowmetry was used to measure neoterminal ileal blood flow in 16 patients who had undergone ileocolonic resection for Crohn's disease and had since remained clinically and biochemically free of disease, and eight control patients who had undergone similar surgery for colonic carcinoma. Four patients with clinically active Crohn's disease of the terminal ileum were also studied. Neoterminal ileal recurrence in those with inactive Crohn's disease was graded endoscopically. The median and minimum of five local blood flow measurements performed in each patient were inversely correlated with the endoscopic recurrence grade (r = -0.52, p = 0.04 and r = -0.63, p = 0.01 respectively). Relative to the control group, median blood flow was non significantly lower in the inactive Crohn's disease group as a whole (p > 0.05) but was significantly reduced in patients with active disease (p = 0.02). A progressive reduction in tissue perfusion may accompany recurrence of Crohn's disease while at a subclinical stage. PMID- 8244139 TI - Comparison of leukocyte excretion and blood loss in inflammatory disease of the bowel. AB - Clinical relapse of inflammatory bowel disease is characterised by increased neutrophil migration into the intestine. The site of the neutrophil chemoattractant(s), whether luminal or mucosal, may be important since, on contact with a chemoattractant, neutrophils cause indiscriminate damage to their immediate surroundings by generating reactive oxygen species and by lysosomal enzyme release. If this happens within the mucosa, inflammation should correlate significantly with tissue damage as assessed by bleeding, but if it occurs within the intestinal lumen, the inflammation would be disproportionately greater than the bleeding such as is seen in classical exudation. Intestinal inflammation and bleeding were quantitated with the simultaneous use of indium-111 labelled neutrophils (four day faecal excretion of indium-111) and chromium-51 labelled red cells in patients with ulcerative colitis (n = 12), Crohn's disease (n = 15), and NSAID induced enteropathy (n = 34). Intestinal inflammation and blood loss correlated significantly (Spearman) in patients with ulcerative colitis (20.3% v 6.5 ml/d (median) r: 0.85, p < 0.001) and NSAiD enteropathy (1.6% v 1.9 ml/d, r: 0.60, p < 0.01) but not in Crohn's disease (17.0% v 2.1 ml/d, r: 0.38, p > 0.1). For a given indium-111 excretion, patients with ulcerative colitis had significantly greater (p < 0.01) blood loss than patients with Crohn's disease. These results suggest that the predominant site of neutrophil chemoattractants may be within the mucosa in ulcerative colitis and NSAID enteropathy and within the lumen in Crohn's disease. PMID- 8244140 TI - Refractory pouchitis: does it reflect underlying Crohn's disease? AB - Typical 'pouchitis' is a well recognised complication of ileal pouches in ulcerative colitis. Infrequently, a refractory pouchitis (RP) presents with certain clinical, endoscopic, and pathological features resembling Crohn's disease and is often ascribed to misdiagnosis of the initial colitis. To test that hypothesis and to identify risk factors for RP, this study reviewed cases of presumed ulcerative colitis with ileal pouches constructed at The Mount Sinai Hospital between 1973 and 1986. Twenty four cases with RP (16 Kock pouches and eight pelvic pouches) and 21 controls were compared for eight clinical variables. The original colectomy slides from 15 RP and 18 control cases were reviewed blindly, classified into five histological categories (corresponding to definite ulcerative colitis, definite Crohn's disease, and three indeterminate groups), and scored for 23 histological features. There were no significant clinical differences between RP and control cases except for more frequent extraintestinal manifestations (38% v 5%) and male preponderance (79% v 43%) in RP. There were also no significant differences between the distributions of RP cases and controls among the five histological categories or in the 23 histological features studied. Refractory pouchitis therefore does not seem to reflect underlying Crohn's disease, but may be linked to immunological mechanisms that are manifested clinically as extraintestinal complications. PMID- 8244141 TI - Hormone replacement therapy prevents bone loss in patients with inflammatory bowel disease. AB - Patients with inflammatory bowel disease have an increased prevalence of osteoporosis, and suffer high rates of spinal bone loss. Hormone replacement therapy (HRT) is effective in the treatment and prevention of osteoporosis but has not been studied in patients with inflammatory bowel disease. A two year prospective study of HRT in inflammatory bowel disease was performed in 47 postmenopausal women aged 44 to 67 years with ulcerative colitis (25) or Crohn's disease (22). Patients had radial and spinal bone density measured annually by single photon absorptiometry and quantitative computed tomography respectively. The mean (95% confidence intervals) annual change in radial bone density was +1.42%/yr (+0.58 to +2.26; P < 0.005) and for spinal bone +2.60%/yr (+1.06 to +4.15; p < 0.005). There was no significant correlation between rates of change of bone density at the two sites, or between the rates of change and the initial bone density either in the radius or spine. Twelve patients were given prednisolone during the study, and their rates of change for spinal bone density were lower, but values were not statistically significantly different from those who did not receive corticosteroids. Changes in bone density for patients with ulcerative colitis and Crohn's disease were not significantly different. The change in bone density did not correlate with the patients' age or number of years after the menopause. It is concluded that HRT is effective in prevention of bone loss in postmenopausal women with inflammatory bowel disease. PMID- 8244142 TI - Prevalence and family risk of ulcerative colitis and Crohn's disease: an epidemiological study among Europeans and south Asians in Leicestershire. AB - The family history of patients identified during incidence studies in Leicestershire were investigated and the prevalence and comparative risks calculated; 1254 patients aged 15 to 80 years were sent a questionnaire about their family history. All cases with a positive family history were reviewed and confirmed cases included in the study. In Europeans the standardised prevalence of Crohn's disease was 75.8/10(5) and that of ulcerative colitis 90.8/10(5). The prevalence of Crohn's disease among South Asians was 33.2/10(5) and that of ulcerative colitis 135/10(5). The prevalence of Crohn's disease in Europeans was significantly greater than that in Hindus (chi 2 = 16, p < 0.001), while the prevalence of ulcerative colitis was significantly lower in Europeans than Hindus (chi 2 = 27, p < 0.001) and Sikhs (chi 2 = 4.4, p < 0.05). The comparative risk of developing ulcerative colitis in first degree relatives of Europeans patients with ulcerative colitis was increased by approximately 15, but the risk of Crohn's disease was not increased. The comparative risk of developing Crohn's disease among first degree relatives of patients with Crohn's disease was increased by up to 35, the comparative risk of ulcerative colitis was approximately 3. The risk among relatives of South Asian patients with Crohn's disease was not increased, but the risk of ulcerative colitis to relatives of patients with ulcerative colitis was. This study supports the view that Crohn's disease and ulcerative colitis arise in people with a genetic predisposition and exposed to some, as yet unknown, environmental factor. PMID- 8244143 TI - Ileal and colonic epithelial metabolism in quiescent ulcerative colitis: increased glutamine metabolism in distal colon but no defect in butyrate metabolism. AB - Previous studies have shown that butyrate is an important energy source for the distal colon, and that its metabolism may be defective in ulcerative colitis (UC). A similar metabolic defect in the ileum might account for the occurrence of 'pouchitis' in UC patients after colectomy. A method has been developed that allows the measurement of metabolism in ileocolonoscopic biopsy specimens, and this has been used to assess butyrate and glutamine metabolism in quiescent UC and controls. Preliminary experiments showed optimal metabolism of butyrate at 1 mmol/l. In controls glutamine metabolism was greater in the ascending (mean (SD)) (4.9 (3.2) nmol/h/micrograms protein) than in the descending colon (1.4 0.7)) (p < 0.05, Mann-Whitney U test), but butyrate metabolism was similar in the two regions (ascending 62.6 (44.2), descending 51.5 (32.0)). Consequently ratios of butyrate/glutamine metabolism were higher in the descending colon (20.6 (14.3)) than in the ascending colon (14.3 (9.6)) (p < 0.05). In UC, rates of butyrate metabolism were similar in the ascending (92.5 (58.3) nmol/h/micrograms protein) and descending (93.3 (115)) colon, and these were not significantly different from controls. In UC, glutamine metabolism was similar in the ascending (6.2 (7.7) nmol/h/micrograms protein) and descending colon (7.8 (7.9)); the metabolism in the descending colon was significantly greater than in controls (p < 0.01). Butyrate (135 (56) nmol/h/microgram protein) and glutamine (24.1 (16.2)) metabolism in the ileum in UC, were not significantly different from control values (butyrate 111 (57), glutamine 15.5 (15.6)). These results confirm that there is regional variation of nutrient utilisation throughout the colon, but they do not support the hypothesis that UC is caused by a deficiency of butyrate metabolism. PMID- 8244144 TI - Omeprazole inhibits colorectal carcinogenesis induced by azoxymethane in rats. AB - Numerous clinical and experimental studies suggest that gastrin plays an important part in the development of colorectal cancer in humans. This study was done to assess the influence of omeprazole induced hypergastrinaemia on the development of colorectal tumours in an experimental animal model. Forty female Sprague-Dawley rats received either omeprazole (40 mumol/kg) or vehicle (0.25% methylcellulose) by once daily oral gavage throughout the experiment. All animals received 12 consecutive weekly subcutaneous injections of azoxymethane (10 mg/kg/week) beginning at week 6. Serum gastrin concentrations were measured during weeks 1 and 5 and at death (week 27). Chronic omeprazole treatment resulted in appreciable hypergastrinaemia during the study, mean gastrin concentrations in omeprazole treated rats being raised by up to nine to 10 fold, compared with vehicle treated control rats (p < 0.001). Despite this, tumour incidence in the omeprazole group was significantly lower at 63%, compared with 95% in the vehicle only group (p < 0.02). The median number of tumours in the omeprazole group (1) compared with the vehicle group (3) was also significantly lower (p = 0.02). Average tumour size, site distribution, and the comparative frequencies of adenomas and adenocarcinomas were similar in the two groups. This study shows that omeprazole protects against colorectal carcinogenesis in this model despite causing appreciable hypergastrinaemia. The mechanism by which this occurs is unclear and merits further investigation. Because of the compounding protective effects of omeprazole, this model is not a suitable one for studying the longterm trophic effects of gastrin on the colon. PMID- 8244145 TI - DNA densitometry of colorectal cancer. AB - DNA analysis was assessed by densitometry for 281 cases of colorectal adenocarcinoma. Detection of aneuploidy in a single case rose from 65% if one, to 92.5% when three or more sections, were analysed. Although aneuploid tumours had significantly larger nuclear areas than near diploid tumours (p = 0.009), densitometric measurements showed no association with clinicopathological variables. DNA content determined by densitometry was compared with that from flow cytometry on 465 tissue sections from 241 cases. Aneuploidy assessed by flow cytometry was significantly associated with that determined by densitometry (p < 0.01 for all comparisons), ploidy state being similar in 381 sections (82%, kappa = 0.63, p < 0.001), and 187 cases (77.6%, kappa = 0.57, p < 0.001). Univariate survival analysis showed that DNA densitometric variables had no significant association with survival in (a) all cases, (b) cases without lymph node metastases, or (c) cases without distant metastases. Multivariate regression analysis of densitometric and clinicopathological variables identified Dukes's stage, patient age, and tumour differentiation as the combination of variables most closely related to survival. Densitometric measurement of DNA content could not significantly improve on the prognostic model containing these three variables. It is concluded that, although the assessment of DNA content by densitometry is comparable with that of flow cytometry, conventional histological variables remain the best predictors of prognosis in colorectal cancer. PMID- 8244146 TI - Ambulatory motility patterns of the transposed short segment colon. AB - Ambulatory manometry and pH-metry were performed on 10 asymptomatic patients who had undergone lower oesophageal replacement with the left hemicolon between two and 20 years previously. Recording of the ambient pH in the intrathoracic colon was carried out simultaneously. In every patient the recording, which lasted approximately eight hours, included upright, supine, prandial, and post-prandial periods. The results were analysed both visually and by computer. The emerging motility patterns showed three basic types of wave, based on amplitudes--low amplitude contractions (LAC) measuring < 60 mm Hg, medium amplitude contractions (MAC) of 60-100 mm Hg, and high amplitude contractions (HAC) of 100-200 mm Hg. Additional subtypes were identified in the LACs and MACs. Using software primarily designed for oesophageal motility, the proportion of propulsive (6% 55%) waves could be distinguished from simultaneous or mixed waves, or both. A proportion of propulsive waves was noted in patients who had undergone surgery 10 or more years previously. It is concluded that the transposed colon retains its normal motility pattern but some adaptation to its new location may occur in the long term. PMID- 8244147 TI - Liquid stool incontinence with severe urgency: anorectal function and effective biofeedback treatment. AB - The motor and sensory function of the anorectum is well characterised in patients with solid stool incontinence. Fewer data are available in the case of liquid stool incontinence. Anorectal sensorimotor function was studied in 16 patients with liquid stool incontinence and severe urgency (10 with diarrhoea) unresponsive to conventional medical treatment, and in 16 healthy volunteers. The only significant difference found between incontinent patients and controls was a reduction in squeeze duration (p < 0.0001). Fourteen patients were selected to receive biofeedback treatment. Treatment was associated with a substantial improvement in continence in 12 patients and with a significant decrease in urgency (p < 0.05). Bowel frequency was not significantly influenced. Most patients showed a persistent improvement in anal motor function. Functional parameters were not predictive of outcome of treatment; the poor responders showed major psychological problems. In conclusion, an anal motor deficit is often present in disabling liquid stool incontinence. Biofeedback may improve anal continence in 75% of patients. PMID- 8244148 TI - Biomechanical wall properties of the human rectum. A study with impedance planimetry. AB - Biomechanical properties of the rectal wall were studied in 17 healthy adult volunteers (nine men and eight women). With impedance planimetry it is possible to obtain simultaneous measurements of pressure and rectal cross sectional area (CSA) during balloon inflations. Rectal distensions were done with an intraluminal balloon using specified pressures up to 40 cmH2O above baseline rectal pressure. Balloon inflation elicited a phase of rapid increase in rectal CSA followed by a phase of slow increase until a steady state was reached. Steady state occurred within 67 to 140 seconds with the shortest period at the highest distension pressures. Steady state rectal CSA values had a non-linear relation to increasing distension pressure. Rectal CSA values in women showed a tendency of being slightly higher than male values at all pressure steps with a significant difference at 3 and 5 cm H2O. Biomechanical parameters were calculated from rectal CSA pressure relations. Circumferential wall tension increased in a linear way. Rectal compliance decreased in a non-linear way with no further decline between 30 and 40 cmH2O. The pressure elastic modulus increased steeply until a distension pressure of 35 cmH2O with no further increase to 40 cmH2O. This suggests that rectal tone is reduced as the muscle fails to resist further distension at 35 cmH2O and higher pressures. Impedance planimetry offers new possibilities for investigation of anorectal physiology through the study of segmental biomechanical wall properties of the human rectum. PMID- 8244149 TI - Randomised trial of targeted chemotherapy with lipiodol and 5-epidoxorubicin compared with symptomatic treatment for hepatoma. AB - Lipiodol injected into the hepatic artery is selectively retained in hepatomas so has been used as a vehicle for cytotoxic drugs. This study compared treatment with 5-epidoxorubicin emulsified in lipiodol and infused into the hepatic artery with symptomatic treatment alone in a randomised trial. Of 136 patients with hepatoma 78 (57%) were not eligible, eight (6%) refused to take part, and 50 entered the trial (chemotherapy: n = 25, symptomatic treatment: n = 25). The two groups had similar prognostic indices. Seven of 25 patients allocated to chemotherapy were unable to receive it. The slight survival disadvantage associated with chemotherapy was not significant (median survival 48 days compared with 51 days, log rank chi 2 = 0.07, p > 0.05). Patients given chemotherapy spent significantly longer in hospital, however (median three days compared with one, p = 0.0008). Changes in symptoms and indices of tumour growth did not differ significantly between the two groups. It is concluded that infusion of 5-epidoxorubicin emulsified in lipiodol for hepatoma increased morbidity but did not affect survival. In addition, most patients were unsuitable for this treatment because of advanced disease. The patients in the trial had a short median survival time so the conclusions may not be valid for other patients with hepatoma. PMID- 8244150 TI - Down regulation of epidermal growth factor receptors in liver proliferation induced by a mixture of triiodothyronine, amino acids, glucagon, and heparin (TAGH). AB - This study investigated the mechanisms by which TAGH solution (a mixture of triiodothyronine, amino acids, glucagon, and heparin) induces DNA synthesis in hepatocytes in the liver of intact rats, with particular reference to events at the epidermal growth factor (EGF) receptor. Both partial hepatectomy and infusion of TAGH stimulated DNA synthesis at 24 hours and both procedures resulted in a reduction of EGF receptors assessed in plasma membranes isolated from rat liver at this time. In cell cultures, while EGF strongly stimulated DNA synthesis and started EGF receptor down regulation, TAGH had only a minor effect (1.5 x basal) on DNA synthesis and did not interact with or down regulate the EGF receptor. Membrane phosphorylation studies, however, showed that TAGH induced phosphorylation of tyrosine residues in the EGF receptor. The in vivo action of TAGH seems to entail recruitment of similar changes in the EGF receptor to those that occur after partial hepatectomy. PMID- 8244151 TI - Postprandial biliary and pancreatic secretion during profound inhibition of gastric secretion in humans. AB - This study assessed the effect of profound inhibition of gastric secretion by an H2 antagonist on postprandial gastric emptying of acid and chyme, and on bile acid and pancreatic enzyme secretion under physiological conditions in humans. Six subjects were studied before and while they were given famotidine (40 mg). This study combined a continuous intestinal perfusion technique using 14C polyethylene glycol (14C-PEG) as duodenal recovery marker, with intermittent sampling of gastric content using PEG 4000 as meal marker. During the three hour study, the area under the curve for gastric acid output decreased from mean (SEM) 88.9 (7.6) mmol for those not receiving treatment, to 21.2 (2.7) mmol for subjects receiving famotidine (p < 0.01). The corresponding values for the rate of acid delivery into the duodenum decreased from 65.2 (11.9) to 16.6 (2.9) mmol (p < 0.05), and those for the rate of gastric emptying of chyme remained unchanged for the group receiving no treatment and during famotidine (1040 (200) v 985 (160) ml respectively, NS). Duodenal bile acid and trypsin output remained unchanged (area under the curve, 457 (128) v 373 (86) umol/kg and 5022 (565) v 5058 (400) IU/kg respectively, NS) receiving no treatment and during famotidine. It is concluded that profound inhibition of postprandial gastric acid secretion by anti-secretory drugs is not accompanied by changes in biliary and pancreatic secretion, mainly because the gastric emptying of chyme is unaffected. PMID- 8244152 TI - Primary culture of bovine gall bladder epithelial cells. AB - Gall bladder epithelial cells serve numerous biological functions and abnormalities in their function are important in the pathogenesis of several gall bladder diseases. Direct studies on cell function are rare due to lack of reliable methods to culture this epithelium. This study reports a reliable and reproducible method of harvesting and culturing gall bladder epithelial cells. Normal bovine gall bladder epithelium, obtained within 20 minutes of slaughter, was rinsed with modified Hanks's balanced salt solution, the mucosa separated and incubated in trypsin--EDTA solution at 37 degrees C. The cells were isolated and resuspended in Dulbeco's modification of Eagles' medium containing 10% fetal calf serum and, after filtration and centrifugation, were plated under aseptic conditions. The growth rate was established by flow cytometry and the morphological characteristics of the growing cells by electron microscopy. Gall bladder epithelial cells grew successfully and visible clusters of cells were present by day two, confluency being reached at 8 to 10 days in collagen coated plates and 12 to 14 days in uncoated plates. Electron microscopy showed typical gall bladder epithelia with microvilli, tight junctions, and mucus droplets. This method proved reliable and reproducible for the culture of gall bladder epithelial cells and should allow direct studies of the biological properties of these cells in human tissue. PMID- 8244153 TI - Inflammatory bowel disease and refractory anaemia (myelodysplasia). AB - Two patients with colitis and refractory anaemia requiring multiple transfusions are described. In one the anaemia was detected at the onset of colitis before drug treatment; in the other it started after 21 years of treatment with sulphasalazine and during an exacerbation of colitis with perianal abscess for which he had just been started receiving prednisolone, cofluampicil and metronidazole. Glomerulonephritis developed two weeks after the onset of anaemia. These findings together with a recent report of four patients with Crohn's disease and refractory anaemia suggests that the association may be more than coincidental and may be immune mediated. PMID- 8244154 TI - Intraperitoneal ivalon mimicking peritoneal malignancy after plugged percutaneous liver biopsy. AB - Intraperitoneal ivalon mimicking peritoneal malignancy after plugged percutaneous liver biopsy is reported in a 20 year old woman with a history of nausea and abdominal distension. PMID- 8244155 TI - Crohn's disease with pulmonary involvement in a 3 year old boy. AB - Extra-intestinal or metastatic Crohn's disease is being recognised increasingly, most commonly in the skin and genitourinary system. Only very rarely has it been reported in the lung. A 3 year old boy who presented with swollen gums and a persistent abnormality on his chest x ray is reported. Lung biopsy specimens showed multiple non-caseating epithelioid granulomas. Subsequent investigation showed the presence of intestinal Crohn's disease. The evidence for abnormal lung function in Crohn's disease and the possible pathogenesis of metastatic pulmonary Crohn's disease are discussed. PMID- 8244156 TI - Hormone replacement therapy in chronic active hepatitis; a case report. AB - There is an increased incidence of osteoporosis in patients with chronic liver disease. Because patients with chronic active hepatitis (CAH) usually require corticosteroids for their liver disease prevention and treatment of bone loss presents a difficult problem. This case report describes a 41 year old female patient with CAH who had a high rate of bone loss. After an early menopause with noticeable menopausal symptoms, she was given transdermal oestrogen replacement therapy. The menopausal symptoms resolved completely, and there was no deterioration in her liver function tests or corticosteroid requirement. In addition, follow up quantitative bone mineral measurements over two years have shown improvement. This case shows the value of measurements of bone density, and oestrogen replacement therapy in CAH, even in the presence of continued corticosteroids. PMID- 8244157 TI - Aldehyde disinfectants and health in endoscopy units. British Society of Gastroenterology Endoscopy Committee. AB - Summary of main recommendations(1) Glutaraldehyde, used in most endoscopy units in the United Kingdom for the disinfection of flexible gastrointestinal endoscopes, is a toxic substance being an irritant and a sensitiser; symptoms associated with glutaraldehyde exposure are common among staff working in endoscopy units.(2) The Control of Substances Hazardous to Health Regulations 1988 (COSHH) obliges the employer to make a systematic assessment of risk to staff of exposure to glutaraldehyde and institute measures to deal effectively with exposure.(3) At present glutaraldehyde remains the first line agent for the disinfection of flexible gastrointestinal endoscopes. Other agents are being developed; a standard means of assessment for flexible endoscope disinfectants should be devised.(4) Equipment and accessories that are heat stable should be sterilised by autoclaving; disposable accessories should be used wherever possible.(5) Flexible gastrointestinal endoscopes should be disinfected within automated washer/disinfectors; trays, bowls or buckets for this purpose are unacceptable.(6) Local exhaust ventilation must be used to control glutaraldehyde vapour. Extracted air may be discharged direct to the atmosphere or passed over special absorbent filters and recirculated. Such control measures must be regularly tested and records retained.(7) Endoscope cleaning and disinfection should be carried out in a room dedicated to the purpose, equipped with control measures to maintain the concentration of glutaraldehyde vapour at a level certainly below the current occupational exposure standard of 0.2 ppm and preferably below the commonly used working limit of 0.1 ppm. Sites other than the endoscopy unit where endoscopy is regularly performed, such as the radiology department, should have their own fully equipped cleaning and disinfection room.(8) COSHH limits the use of personal protective equipment to those situations where other measures cannot adequately control exposure. Such equipment includes nitrile rubber gloves, apron, chemical grade eye protection, and respiratory protective equipment for organic vapours.(9) Monitoring of atmospheric levels of glutaraldehyde should be performed by a competent person such as an occupational hygienist; the currently preferred method of sampling uses a filtration technique, the commercially available meters being less reliable.(10) Health surveillance of staff is mandatory; occupational health records must be retained for 30 years.(11) Endoscopy staff must be informed of the risks of exposure to glutaraldehyde and trained in safe methods of its control. Only staff who have completed such an education and training programme should be allowed to disinfect endoscopes.(12) The unsafe use of glutaraldehyde has significant health and legal consequences; the safe use of glutaraldehyde may have revenue consequences that contribute significantly to the cost of gastrointestinal endoscopy. PMID- 8244158 TI - Colonic epithelial metabolism in ulcerative colitis. PMID- 8244159 TI - Helicobacter pylori positive resistant duodenal ulcers. PMID- 8244160 TI - Helicobacter pylori positive resistant duodenal ulcers. PMID- 8244161 TI - Helicobacter pylori positive resistant duodenal ulcers. PMID- 8244162 TI - The physician and leadership challenges for the 1990s. PMID- 8244163 TI - Ifosfamide and mesna in epithelial ovarian carcinoma. AB - Ifosfamide is a cyclophosphamide analogue synthesized in the 1960s with antineoplastic activity demonstrated in early broad-ranging phase I studies conducted in Germany in the 1970s. Because of significant urothelial toxicity, phase II studies in ovarian cancer in this country were delayed until the urinary epithelial protector mesna became available in 1985. Since that time, two well executed prospective trials have shown that this agent produces measurable responses in about 20% of women with ovarian epithelial cancer recurring after primary chemotherapy and in 12% of those with tumors refractory to first-line therapy with regimens including cisplatin. Toxicity includes moderate to severe hematologic toxicity, renal dysfunction which is usually reversible, and CNS abnormalities including lethargy, somnolence, and disorientation. The risk of toxicity may be increased in patients with compromised hepatic or renal function and in those with hydronephrosis or hypoalbuminemia. PMID- 8244164 TI - Hexamethylmelamine/altretamine as second-line therapy for epithelial ovarian carcinoma. AB - The purpose of this report was to review second-line hexamethylmelamine (HMM) chemotherapy of epithelial ovarian cancer to determine if HMM was active in cisplatin-resistant disease. Forty-four women with measurable disease received 100-300 mg/day HMM for 14 days, courses repeated every 4 weeks. There were 6 complete and 3 partial responses for an objective response rate of 20%. Among responding patients disease-free survival was 55% and overall survival was 88% at 3 years. Five of the 6 patients with a complete response remained disease-free at 10-117 months. Only 7/35 (20%) nonresponding patients were alive with mean follow up of 16 months, and all had persistent cancer. Five women manifesting disease progression during cisplatin or carboplatin were subsequently treated with HMM, and none responded. Seventeen patients developing progressive cancer while receiving HMM were subsequently treated with cisplatin or carboplatin and objective responses occurred in 5 (29%). HMM was an active drug against epithelial ovarian cancer previously treated with cisplatin, but further study is needed to determine its activity against cisplatin-resistant ovarian cancer. PMID- 8244165 TI - Biologic therapy in the management of refractory ovarian cancer. AB - The success of platinum-based chemotherapy in the initial management of advanced stage ovarian cancer has been challenged by the emergence of drug-resistant tumors in the majority of patients. Even taxol, which can achieve clinical responses in one-third of patients with platinum-resistant tumors, may fail to substantially alter long-term survival for many patients with refractory disease. Thus, there continues to be a need for alternative therapeutic strategies. Specific roles for primary or adjunctive biologic therapy in the management of refractory ovarian cancer have not yet been defined. For example, a variety of conjugated monoclonal antibodies have been evaluated for over 10 years, but have not yet been shown to be effective with acceptable levels of host toxicity. The closest candidate for standardized biologic therapy is intraperitoneal interferon for treatment of microscopic residual disease. Adjunctive hematopoietic colony stimulating factors are widely used with taxol to achieve greater dose intensity and cumulative drug delivery, although they have not yet been shown to improve response or survival. Appreciation of the complex pathways regulated by growth factors and cytokines will provide a more challenging framework for continued development of biological therapeutics in the future. PMID- 8244166 TI - Screening for ovarian cancer: the preliminary experience of a familial ovarian cancer center. AB - Women with a family history of ovarian cancer represent a high-risk group for the development of epithelial ovarian cancer. From July 1990 through December 1992, 386 women with a first-degree or multiple second-degree relatives with confirmed ovarian cancer were enrolled in a study to assess the utility of screening with transvaginal sonography, color flow doppler, and CA125. The mean age of the group was 41; 85% were premenopausal and 89/384 (23%) had 2 or more relatives with ovarian cancer. An initial ultrasound examination was abnormal in 89/384 (23%), 89% of whom were premenopausal. A persistent ovarian mass was detected in 15 patients and all were surgically proven to be benign. Mean CA125 levels were significantly higher and more variable in pre- vs postmenopausal women. CA125 was > or = 35 U/ml in 42/386 (11%) (36-232 U/ml). All but one of these women were premenopausal and 50% subsequently normalized. Two patients who were surgically explored for a rising CA125 had normal ovaries. An additional 19 patients have undergone prophylactic oophorectomy with no consistent histopathologic abnormality identified. These data demonstrate the difficulty inherent in screening a predominantly premenopausal population and do not clearly establish the efficacy of these modalities in the early detection of ovarian cancer. PMID- 8244167 TI - Secondary cytoreduction--thoughts on the "pro" side. AB - The advantages of secondary cytoreduction in the treatment of women with ovarian carcinoma include improved survival and potential improvement in quality of life. To summarize the literature on this topic, patients can be grouped into four distinct clinical settings. Debulking at the time of second-look laparotomy, interval debulking following three to four courses of chemotherapy, and debulking at the time of recurrence following a disease-free interval are scenarios which offer clinical advantages to women with ovarian carcinoma. However, there is little or no evidence to support a survival or quality of life advantage for women who undergo secondary debulking despite progressive disease on primary platin-based chemotherapy. Data on the pro side of the secondary tumor debulking debate are summarized here. PMID- 8244168 TI - Secondary cytoreduction in ovarian cancer: pro or con? AB - During a symposium in the therapy for ovarian cancer as a prelude to the Gynecologic Oncology Group meeting in July 1992, the relative merits of secondary cytoreductive surgery were debated. The author presented part of the con argument. The content of that position is presented. PMID- 8244169 TI - Advanced ovarian carcinoma: molecular evidence of unifocal origin. AB - The clonal origin has sometimes been disputed in advanced epithelial cancers when multiple sites of peritoneal tumors were present because of the developmental similarity of the surface epithelium of the ovary and the peritoneal mesothelium. We have taken two independent approaches to study the clonality of advanced epithelial carcinoma. Tumor samples of both ovaries, and/or omentum, and/or peritoneum from nine patients were examined. The analysis of loss of heterozygosity (LOH) at 86 polymorphic chromosomal loci distributed on every chromosome revealed the loss of the same allele in every tumor of each patient when LOH was detected. In addition, we used a restriction-fragment-length polymorphism in an X-linked sequence (DXS255) to examine the pattern of X chromosome inactivation based on the differential methylation of DXS255 in active and inactive X chromosomes. Inactivation of the same X chromosome in various tumors from the same patient was observed in each of the five informative patients. Our results strongly suggest that the advanced epithelial cancers are monoclonal in origin. PMID- 8244170 TI - Early invasive carcinoma of the cervix. AB - Ninety-two patients with early invasive carcinoma of the cervix (5 mm or less) treated between July 1977 and June 1990 are reviewed. Eighty patients had squamous cell carcinomas and 12 had adenocarcinomas. The diagnosis was established by conization in 77 of 92 (83.6%) patients. Thirty-six patients (39%) had a depth of stromal invasion of 1 mm or less, 32 patients (35%) between 1 and 3 mm, and 24 patients (26%) between 3 and 5 mm. Forty-four patients were treated with radical hysterectomy and bilateral pelvic lymphadenectomy (RHND). None of these patients had positive lymph nodes. Thirty-three patients were treated with conservative hysterectomy (CH), 4 with modified radical hysterectomy, and 2 with trachelectomy. Six patients received radiotherapy. Three patients were treated by conization only. Two patients developed in situ carcinoma (CIS) of the vagina 12 months after CH for lesions on conization that invaded less than 1 mm. In both cases the cone margins were positive, and in one a microscopic focus of CIS of the cervix was present at the resection margin of the hysterectomy specimen. A third patient developed an invasive lesion of the vagina 25 months after CH for a lesion that invaded 2.5 mm in a cone whose margins were not specified, but the hysterectomy margins were clear. All 3 patients were successfully retreated. The remaining patients are free of disease for a median follow-up of 51 months. The results of the study indicate that CH is adequate therapy for patients in whom the diagnosis of early invasive cervical cancer is established by conization with free margins and the depth of invasion is 3 mm or less. Although only 1 of 24 patients with invasion > 3 mm but < or = 5 mm had a CH, pathologic findings in 18 patients who had RHND suggest that CH would have been sufficient for these since there were no instances of spread to nodes or parametrium. PMID- 8244171 TI - Laparoscopically assisted surgical staging (LASS) of endometrial cancer. AB - We report 59 patients who were considered candidates for laparoscopically assisted surgical staging (LASS) to manage their clinical stage I adenocarcinoma of the endometrium. Their ages ranged from 40 to 85 years, with a mean of 69; their weights ranged from 102 to 267 pounds, with a mean of 153 pounds. Patients with intraperitoneal disease were taken off study. Laparoscopic pelvic and para aortic lymphadenectomies were performed based on the grade of the tumor and the depth of myometrial invasion. Six patients were discovered to have intraperitoneal disease. Of the remaining 53 patients, 29 underwent lymphadenectomy, 1 of whom had positive para-aortic nodes. Of the 24 patients who did not have laparoscopic lymphadenectomy, 2 should have, according to the study criteria; however, obesity precluded this from being performed. Eight patients had grade 3 lesions; of these, 4 lesions had metastasized. The remaining 3 patients with metastatic disease had grade 2 lesions. Complications were related to the laparoscopically assisted vaginal hysterectomy and resulted in two laparotomies: one for a transected ureter and the other for a cystotomy. Estimated blood loss was < 200 cc and the average hospital stay was 2.9 days. We feel that LASS is an attractive alternative to the traditional surgical approach in patients with stage I endometrial carcinoma. PMID- 8244172 TI - Differences in the morbidity of radical hysterectomy between gynecological oncologists. AB - A multivariate analysis was performed on 405 patients who underwent radical hysterectomy and pelvic lymphadenectomy by eight surgeons for stage IB cervical carcinoma, to determine the influence of primary surgeon on morbidity. Patient characteristics analyzed (mean/proportion) were age (41 years), quetelet index (25.4), American Society of Anesthesiologists classification of physical status (0.5% > 2), previous laparotomies (23%), previous radiation (0.7%), prophylactic antibiotics (95%), prophylactic heparin (67%), tumor size (1.0 cm), histology (68% SCC), grade (68% grades 2 or 3), vascular space involvement (45%), pelvic lymph node metastases (6%), and depth of invasion (6.6 mm). Morbidity characteristics analyzed (mean/proportion) were blood loss (910 ml), operative time (3.0 hr), intra-op complications (5%), post-op infectious (21%) and non infectious complications (7%), transfusions (35%), post-op hospital stay (9.9 days), time to normal urine residual (9.0 days), and bladder dysfunction at 3 months post-op (21%). Mean tumor size was the only preoperative characteristic that was significantly different among surgeons (P < 0.001). Of the factors evaluated for morbidity, mean blood loss (P < 0.0001), operative time (P < 0.001), and postoperative hospital stay (P < 0.001) varied among physicians as did the incidence of blood transfusion (P < 0.0001) and bladder dysfunction at 3 months postoperatively (P < 0.0001). On multivariate analysis, surgeon was independently significant for blood loss (P < 0.0001), operative time (P < 0.0001), postoperative hospital stay (P < 0.001), incidence of blood transfusion (P < 0.0001), and bladder dysfunction at 3 months postoperatively (P < 0.0001). Despite differences in tumor size, patients appeared similar among the surgeons. Differences in patient morbidity among surgeons do exist and are of significant magnitude. Since the design of surgical trials to assess the therapeutic ratio should include not only measures of efficacy, but also measures of morbidity to be meaningful, intersurgical morbidity between centers/surgeons must continue to be quantified. PMID- 8244173 TI - Can ovarian masses be characterized using ultrasound? PMID- 8244174 TI - To drain or not to drain: a retrospective study of closed-suction drainage following radical hysterectomy with pelvic lymphadenectomy. AB - The records of 115 patients were reviewed following radical hysterectomy with pelvic lymphadenectomy for early stage (IA2 to IIB) cervical cancer to determine the effectiveness of closed-suction drainage in decreasing postoperative morbidity. In Group 1 (n = 67), closed-suction (Jackson-Pratt) drains were placed bilaterally in the pelvic lymphadenectomy sites, whereas in Group 2 (n = 48), no drains were utilized. The groups were similar for mean age, tumor histology, disease stage, and preoperative serum albumin and total protein. There was no difference in the mean operative time, mean estimated blood loss, or transfusion rate. Likewise, febrile morbidity rates, incidence of pelvic cellulitis, length of postoperative ileus, and total hospital stay were similar. Group 1 had an increased rate of rehospitalization and morbidity directly related to the presence of the drains. The data suggest that prophylactic surgical drainage may not be necessary to prevent postoperative morbidity following radical hysterectomy with pelvic lymphadenectomy and on occasion contributes to postoperative complications. PMID- 8244175 TI - Accuracy of frozen section for lymph node metastasis in patients undergoing radical hysterectomy for carcinoma of the cervix. AB - The efficacy of frozen section in detecting metastases in pelvic and or periaortic lymph nodes during radical hysterectomy is unknown. The finding of positive nodes may result in termination of the operative procedure. In this study, we attempted to determine the accuracy of frozen sections in this situation. Intraoperative pathology consultation records were examined for 127 patients undergoing surgical exploration for radical hysterectomy between 1977 and 1992. Microscopic slides of lymph nodes were reviewed for accuracy. Metastasis diameters were measured and blocks cut five close microtome levels deeper. In 19 cases (15%) positive nodes were documented on permanent section, with metastases ranging in size from less than 1 to 19 mm. Thirteen cases of node metastasis were diagnosed at frozen section. All were suspicious to the pathologist on palpation and gross inspection after bisection. Six cases were missed by sampling error on frozen section; in 4, metastases were smaller than 1 mm; in 1, between 2 and 3 mm; and in 1, 19 mm. The sensitivity was 68%, the false negative rate was 32%, and the specificity was 100%. No cases were false positive at frozen section. The frequency of nodal metastasis and detection rate by frozen section did not differ significantly between carcinoma types. No micrometastases (< 2 mm) were detected by frozen section. All micrometastases were no longer present within five microtome levels. Frozen section diagnosis of pelvic node metastasis is a highly specific procedure which should alter intraoperative management of early-stage cervical cancer. PMID- 8244176 TI - Biodistribution and intraoperative evaluation of radiolabeled monoclonal antibody MX35 in patients with epithelial ovarian cancer. AB - Murine monoclonal antibody (MAb) MX 35 shows strong homogeneous reactivity with more than 90% of epithelial ovarian cancers. Twenty-five patients with advanced ovarian cancer were entered into a clinical trial using 125I- or 131I-labeled MX 35 in doses of 2, 10, or 20 mg administered by intravenous (i.v.) or intraperitoneal injection. All patients underwent laparotomy at 7 to 20 days following MAb injection to assess tumor distribution, obtain biopsies of tumor and normal tissue, and evaluate the use of an intraoperative hand-held gamma detecting device. Following i.v. injection, serum Mab half-life was 36 hr. Tumor biopsies obtained at surgery showed MAb accumulation of from 6.7 x 10(-3) to 4.0 x 10(-5)% injected dose/g of tissue. There was no correlation between absolute MAb accumulation in tumor and MAb dose administered. Regression analysis showed a correlation between MAb accumulation and the interval between MAb injection and surgery (P = 0.008). Specific localization of MAb in tumor was demonstrated by tumor:normal tissue ratios ranging from 2.3:1 to 34:1 (mean, 10.18:1). The tumor:normal tissue ratios were not significantly related to MAb dose, the level of immunohistochemical antigen expression, or the interval between MAb injection and surgery. Due to the relatively long serum half-life, mean tumor:serum ratios were only 1.53 following IV injection. This ratio did not correlate with MAb dose, days from injection, or antigen expression. There was an excellent correlation (P = 0.001) between MAb uptake, as measured by the intraoperative hand-held gamma counter, and direct gamma counting of excised tissues. MAb MX 35 localizes well to tumor in selected patients with ovarian cancer, and MAb uptake can be reliably quantitated in vivo with the hand-held intraoperative gamma counter. PMID- 8244177 TI - High-dose-rate brachytherapy in primary stage I and II vaginal cancer. AB - Thirteen patients with primary vaginal cancer were treated with external beam irradiation and high-dose-rate brachytherapy. Median age was 65 years old. Five tumors were stage I, 4 stage IIA, and 4 stage IIB. Twelve tumors were squamous and 11 were moderately or poorly differentiated. Median tumor diameter was 4 cm. Patients were treated with external beam irradiation (4500 cGy) and high-dose rate brachytherapy (2000-2800 cGy in three to four fractions). All 13 patients had a complete response. Local control was achieved in 12 patients (92%) at a median follow-up of 2.6 years. No acute or chronic intestinal or bladder grade 3 or 4 toxicity was observed. Moderate to severe vaginal stenosis occurred in 6 patients (46%). Treatment of stage I and II primary vaginal cancer with external beam irradiation and high-dose-rate brachytherapy appears to produce a high response rate, local control, and survival with minimal complications. PMID- 8244178 TI - A morphology index based on sonographic findings in ovarian cancer. AB - A morphology index based on morphologic characteristics of ovarian tumors was developed. Specific categories included tumor volume, wall structure, and septal structure. A point scale (0-4) was developed within each category with the total points per evaluation varying from 0-12. Sonograms on 121 patients undergoing exploratory laparotomy for ovarian masses were evaluated using this index. Eighty ovarian tumors had a morphology index score < 5, and all were benign (negative predictive value, 1.000). In postmenopausal patients, a morphology index score > or = 5 had a positive predictive value for malignancy of 0.45. All ovarian malignancies had significant abnormalities in wall structure and all had a total volume in excess of 10 cm3. The findings of the present investigation indicate that the morphology index is a cost effective adjuvant method which significantly increases the specificity and positive predictive value of transvaginal sonography. The routine application of a morphology index to screening sonography should decrease the amount of diagnostic surgery performed in order to detect each case of ovarian cancer. PMID- 8244179 TI - Phase I and pharmacokinetic study of intraperitoneal ormaplatin. AB - Ormaplatin is a cisplatin analog which has demonstrated activity against cisplatin-resistant tumors in preclinical studies. We delivered 28 cycles to 14 patients in a phase I trial of intraperitoneal ormaplatin given every 28 days. The maximum tolerated dose was 88.4 mg/m2 and acute dose-limiting toxicity was abdominal pain. Other toxicities include nausea, emisis, fever, and severe neuropathy seen in 1 patient at a cumulative dose of 399 mg/m2. No objective responses were observed. Hematologic toxicity was mild. The dose recommended for future trials of intraperitoneal ormaplatin is 66.5 mg/m2. Pharmacokinetic analysis performed at a dose of 66.5 mg/m2 demonstrated that the initial phase of elimination from the peritoneal cavity follows first-order kinetics with k = 0.69 hr-1 and half-life of 1.4 hr. Plasma pharmacokinetic behavior is best described by biexponential model with k1 = 0.369 hr-1, k2 = 0.107 hr-1, and first half-life of 2.9 hr and second half-life of 8.4 hr. Pharmacologic advantage, calculated by ratio of peritoneal to plasma AUC, is 17.1. If site-specific activity is demonstrated, then the intraperitoneal route of administration of ormaplatin at 66.5 mg/m2 may be beneficial. PMID- 8244180 TI - Salvage therapy in ovarian cancer: is there a role for intraperitoneal drug delivery? AB - Phase 2 trials employing several antineoplastic agents have demonstrated that surgically documented responses, including complete responses, can result when patients with ovarian cancer are treated by the intraperitoneal route in the salvage setting following initial systemic chemotherapy. Activity is essentially limited to patients with very small-volume residual disease (microscopic cancer or largest visible tumor masses < or = 0.5 cm in maximum diameter) when the salvage regional therapy program is initiated. In the absence of a randomized phase 3 trial, the ultimate impact of these surgically documented responses on survival is difficult to evaluate, although long-term disease-free survival (> 4 years) has been demonstrated following intraperitoneal therapy in this clinical setting. PMID- 8244181 TI - Clinical reversal of drug resistance in ovarian cancer. AB - Human ovarian cancer cell lines and relevant in vivo model systems have been used to identify mechanisms of resistance associated with alkylating agents and platinum compounds. Drug resistance in ovarian cancer is multifactorial and it is probable that additional mechanisms than those already identified may be responsible for clinical drug resistance. Presently drug transport, increased inactivation in the cytosol, increased repair of damaged DNA, and alterations in signal transduction pathways have been shown to account for resistance of alkylating agents and platinum compounds. Clinical trials have been initiated with agents such as buthionine-sulfoximine, an inhibitor of glutathione biosynthesis which decreases the ability of resistant cells to inactivate platinum compounds prior to their interaction with DNA. In a phase I trial, it has been demonstrated that glutathione levels can be depleted both in normal tissues and tumor biopsies from drug-resistant ovarian cancer patients. Additional clinical trials are needed to determine the effect that glutathione reduction has upon response rate to alkylating agents and platinum compounds. Ultimately, successful modulation of drug resistance may require a combination of agents which inhibit multiple critical biochemical sites. In addition, an understanding of the mechanisms associated with antineoplastic drug resistance may lead to novel therapeutic strategies aimed at preventing the emergence of clinically relevant resistance. PMID- 8244182 TI - Is there a role for consolidation or salvage radiotherapy after chemotherapy in advanced epithelial ovarian cancer? AB - Cure rates following surgery and platinum-containing chemotherapy in advanced epithelial ovarian cancer are only 20-30%. Relapse rates even after complete chemotherapy responses or those leaving only microscopic residual disease are disappointingly high at 30 to 80%. Strategies to improve outcome, consolidate the results of surgery and chemotherapy, or salvage those with residual disease include whole-abdominopelvic irradiation. This paper reviews its usage to date in 28 trials. The results of sequential therapy are generally disappointing, possibly because of inappropriate patient selection, toxic regimens, underlying biological factors, and difficulties interpreting the uncontrolled studies. Tumor residuum appears to be one factor predictive of survival: no residuum, 76%; microscopic or < 5 mm, 49%; macroscopic, 17%. Other factors may be tumor grade and patient age. Appropriate selection of patients for controlled studies of sequential radiotherapy after surgery and chemotherapy include those with negative second-look laparotomy who had large residual Stage III presentations prechemotherapy, are over 50 years of age, or who have grade 3 tumors. Others are those with microscopic residual disease, grade 1 or 2. If therapy is to be successfully completed with minimal morbidity, abdominal radiotherapy should be limited to < or = 25 Gy, initial chemotherapy to six courses, and surgery to initial debulking and second-look laparotomy. PMID- 8244183 TI - Advancement of meiotic resumption in graafian follicles reduces fertility in the rat. AB - Fertile life of oocytes is usually considered to be related to ovulation time. In the present study, fertile life of rat oocytes was studied in relation to resumption of meiosis. In pro-oestrus, meiotic resumption without concomitant ovulation was induced in most graafian follicles by injection of a small amount of LH or FSH followed by Nembutal. These follicles either ovulated or developed into luteinized unruptured follicles if Ovalyse (a GnRH analogue) was given 8 h after LH or FSH. In subsequent experiments, rats were injected with FSH or saline, and Nembutal; 4 or 8 h later, Ovalyse was given to induce ovulation; the rats were mated 14 h after Ovalyse. At day 20 of pregnancy, fetal survival was 30% in rats with meiosis advanced by 8 h, against 91% and 70% in rats advanced by 0 or 4 h, respectively. Mortality occurred mainly during pre-implantation and early post-implantation. Advanced resumption of meiosis may cause pre-ovulatory ageing of oocytes; consequently, viability of these oocytes after ovulation is reduced. PMID- 8244184 TI - Origin of first trimester 17-hydroxyprogesterone levels as determined in pregnancies by donor oocyte fertilization. AB - The study presented herein measured 17-hydroxyprogesterone (17-OHP) levels in women with ovarian failure who conceived by transfer of embryos which resulted from donor oocytes fertilization. A significant increase in 17-OHP during the first trimester was seen compared to baseline nonpregnant levels. The 17-OHP levels increased from a baseline average of 47.7 +/- 9.7 ng/dl to a first trimester average of 175.8 +/- 80.6 ng/dl in the donor oocytes recipients vs. 63.0 +/- 38.0 ng/dl baseline to 295.0 +/- 83.9 ng/dl first-trimester in the control group. Initially these data may appear to contradict previous findings demonstrating a lack of 17-OHP secretion by the first-trimester placenta. However, by comparing the first-trimester progesterone (P) levels of normal pregnant women, and also measuring 17-OHP in patients with natural menopause and surgical menopause given exogenous P we concluded the following about the origin of first-trimester sera 17-OHP levels: hydroxylation of P to 17-OHP by the ovaries, some secretion by the first trimester placenta; and also increased adrenal conversion of P to 17-OHP. Contributing to the total serum 17-OHP level is the fact that there is cross-reactivity with P to 17-OHP. PMID- 8244185 TI - Changes in age distribution of erythrocytes during pregnancy: a longitudinal study. AB - This prospective study was undertaken to evaluate age distribution of erythrocytes longitudinally throughout pregnancy. Serial estimates of density distribution of cells (DDC), reticulocyte count, red cell distribution width (RDW), and mean corpuscular volume (MCV) in a group of healthy pregnant women at 12, 16, 20, 24, 28, 32 and 36 weeks gestation and during the latent phase of labor (39.04 +/- 1.5 weeks). MCV increased constantly during pregnancy; however, the increase was not statistically significant. The mean reticulocyte percentage rose significantly at the 28th week, thereafter it remained at high levels until delivery. RDW increased significantly at the 20th week, remained at high levels until the 32nd week and thereafter declined towards the delivery. The cumulative DDC curve gradually shifted to the right during pregnancy, indicating a less dense cell population as the pregnancy progressed. In conclusion, a continuous alteration in age distribution of erythrocytes towards a younger cell population takes place from early pregnancy until the delivery of the fetus. PMID- 8244186 TI - Elevated plasma interleukin 6: a biochemical marker of human preterm labour. AB - Maternal peripheral venous plasma interleukin 6 (IL-6) concentrations were determined during human pregnancy and labour at term and preterm. During preterm labour, IL-6 concentrations were significantly elevated (p < 0.02) compared to gestationally matched, non-labouring controls (53.7 +/- 15.7 pg/ml, n = 17, and 15.4 +/- 6.4 pg/ml, n = 23, respectively). IL-6 concentrations did not vary significantly during normal pregnancy and labour at term. These data support a role for IL-6 in the pathogenesis of human preterm labour, are evidence for the contention that preterm labour is mechanistically distinct at a biochemical level from normal labour at term and identify maternal peripheral venous plasma IL-6 as a biochemical marker of this condition. PMID- 8244187 TI - Venous placental reactivity to serotonin in normal and preeclamptic gestants. AB - The effect of serotonin (5-HT) on the contractile activity of venous chorionic rings from normal and preeclamptic gestants was analyzed. The results indicate that the resting tension did not differ between the groups, but the spontaneous rhythmic contractions began earlier and were of higher amplitude in the preeclamptic group. Dose-response data indicate that venous segments from preeclamptic patients developed a 1.7-fold increase in sensitivity to 5-HT. These effects were accompanied by an increase in maximal isometric tension (Emax) and relaxation times. Decreased desensitization and lack of tachyphylaxis was observed in most of the vascular preparations from the preeclamptic group. These data suggest that 5-HT may have a role in the modulation of preeclampsia. PMID- 8244188 TI - Pregnancy-induced hypertension is associated with changes in maternal and umbilical blood antioxidants. AB - Seventeen pregnancies with pregnancy-induced hypertension (PIH) and 28 control pregnancies were analyzed with regard to maternal and fetal blood antioxidants and lipid peroxidation products (conjugated dienes). In PIH, maternal blood levels of conjugated dienes were higher than in normal pregnancy. Also the activities of platelet and plasma glutathione peroxidase (GSHPx) were higher in PIH. In umbilical cord blood, the appearance of conjugated dienes, the concentration of vitamin E and the activity of erythrocyte GSHPx were lower than the corresponding maternal values. There was no difference between PIH and normal pregnancy in the appearance of conjugated dienes in cord blood, but erythrocyte GSHPx and plasma vitamin A were lower in PIH. Cord blood plasma vitamin A showed a negative correlation to maternal mean arterial pressure. We suggest that lipid peroxidation is involved in the pathogenesis of maternal PIH, and it may also have effects on the vascular function and antioxidant status of the fetus. PMID- 8244189 TI - Abnormal embryogenesis in the etiology of ectopic pregnancy. AB - Flow cytometric measurements of DNA using paraffin-embedded retrospective material were performed on 42 samples of ectopic conceptuses to see whether there were any aberrations from normal DNA. It was found that DNA aneuploidy was associated with tubal implantation in 14 (33%) cases. The range of DNA indices of the DNA aneuploid peaks was from 1.14 to 2.17. Near-diploid karyotype changes were seen in 8 (19%) cases. Six cases were found to be tetraploid. In 18 abortions with known cytogenetic status, all 3 cytogenetically normal cases showed DNA diploidy in flow cytometry. It was also found that 3 out of 10 cases of trisomy 21 were DNA aneuploid. Flow cytometric DNA reflects only chromosomal changes where a significant number of cells have either lost or acquired several chromosomes. DNA aneuploidy of ectopic pregnancies suggests a grave chromosomal aberration in one third of the cases. Abnormal embryogenesis may contribute significantly to the occurrence of ectopic implantation. PMID- 8244190 TI - Age distribution of erythrocyte population in women with twin pregnancy. AB - An evaluation of the age distribution of erythrocytes in twin gestation was performed. Based on previous studies on age distribution of erythrocytes in late pregnancy, we have proposed that a more pronounced shift in age distribution of erythrocytes in twin pregnancy may be expected. We have used the density distribution of cells method for evaluation of age distribution of erythrocytes in women with twin pregnancies. Control curves for pregnant and nonpregnant women were obtained from our recently published studies. The cumulative density distribution of cells curve of erythrocytes of women with twin gestation was evidently shifted to the right as compared with the curve of erythrocytes of women with single gestation. The shift in age distribution of erythrocytes may suggest that erythrocytes in women with twin gestation have a shorter life span as compared to the life span of erythrocytes in women with singles. The age distribution shift of erythrocytes in women with twin gestation may be attributed to human placental lactogen augmentation of erythropoietin action. PMID- 8244191 TI - Contractile effects of endothelin 1 and endothelin 3 on myometrium and small intramyometrial arteries of pregnant women at term. AB - This in vitro study was undertaken to investigate the effects of endothelin 1 (ET 1) and endothelin 3 (ET-3) on intramyometrial arteries and myometrial tissue of the uterus from pregnant women at term. Segments of small myometrial arteries (outer diameter 100-500 microns) obtained at cesarean section were isolated and mounted in tissue chambers. The effect of ET-1 and ET-3 on isometric tension was compared with the action of noradrenaline. In similar experiments on myometrial strips, the effect of ET-1 and ET-3 was studied in comparison with oxytocin. ET-1 and ET-3 both produced contractions of arteries and myometrial strips, ET-1 being the most potent. The response of ET-1 was more powerful than the effect of the reference compounds, i.e. noradrenaline with regard to myometrial arteries and oxytocin with regard to the myometrium. It is concluded that endothelins are powerful constrictors of myometrial arteries from pregnant women and possess strong oxytocic effects. In particular, ET-1 could be involved in the regulation of myometrial contractility and/or uteroplacental blood flow. PMID- 8244192 TI - Serum laminin levels in normal pregnancy and preeclampsia. AB - Laminin is a large noncollagenous glycoprotein localized in the trophoblast and glomerular basement membrane. We measured laminin levels in maternal serum, umbilical cord serum and amniotic fluid, both from preeclamptic and normal pregnant women, by enzyme immunoassay. The serum laminin levels in the preeclamptic group were significantly (p < 0.05 to p < 0.01) higher than those in the normal pregnant group. It has been suggested that laminin plays an important role in implantation of the placenta during early pregnancy. In this study, the amniotic-fluid laminin level at term was found to be lower than maternal serum laminin. We postulate that laminin may not have an important role in the maintenance of late pregnancy. There was a significant positive correlation between maternal serum laminin levels and serum uric acid levels. There was no significant correlation between maternal serum laminin level and blood pressure, urinary protein concentration, or any other laboratory data. These results suggest that there is damage of glomerular and placental spiral arteries in preeclampsia. PMID- 8244193 TI - Hormone anabolic/catabolic balance in female endurance athletes. AB - The anabolic/catabolic hormone balance in 13 female endurance athletes was studied during basal conditions by comparing their serum concentrations of androgenic and catabolic steroids, growth hormone, insulin-like growth factor 1 (IGF-1) and insulin with corresponding values in 15 matched sedentary controls. Higher cortisol and lower levels sex hormone-binding globulin (SHBG) were found in the athletes, and these differences were enhanced depending on the degree of menstrual disorders. There was a negative correlation between cortisol and SHBG levels. No differences were found in concentrations of androgens, IGF-1 and insulin. The results indicate a normal anabolic activity in oligomenorrheic athletes, but due to their distinct hypercortisolism, the anabolic/catabolic hormone balance is changed to a predominantly catabolic state. PMID- 8244194 TI - Metronidazole combined with nystatin (vagitories) in the prevention of bacterial vaginosis after initial treatment with oral metronidazole. AB - In a double-blind trial comprising 66 patients we assessed the effect of metronidazole-nystatin vagitories on the prevention of bacterial vaginosis (BV) in women using IUD as a contraceptive method after an initial oral single dose of 2.0 g metronidazole and 7 days of intravaginal metronidazole-nystatin or placebo treatment. The prophylactic treatment consisted of metronidazole-nystatin or placebo vagitories applied at bedtime for 3 days after menstruation over 6 consecutive menstrual periods. The patients were randomized in two study groups: a treatment group of 32 patients (group A) and a placebo group of 34 patients (group B). The overall objective cure rate after the initial treatment was 97% in group A and 91% in group B. After 6 months of follow-up, the overall cumulative objective cure rate in group A was 100%, and 76% in group B. The single-dose oral treatment was well tolerated and no notable side effects were recorded. PMID- 8244195 TI - Human papilloma virus infection and Ki-ras oncogene in paraffin-embedded squamous carcinomas of the cervix. AB - 42 paraffin-embedded squamous cervical carcinomas were screened for the presence of human papilloma virus (HPV; 6b, 11, 16, 18) and for activation of the Ki-ras oncogene family by polymerase chain reaction. In 72% of cases we found one or more HPV types, but no mutations of the Ki-ras gene (codon 12-1 and 61-1, 61-2 and 61-3) were found. We conclude that mutations of the Ki-ras oncogene, at the positions analyzed, are not likely to be involved in the events leading to cervical carcinogenesis. PMID- 8244196 TI - Primary Langerhans cell histiocytosis of the vulva. AB - Primary Langerhans cell histiocytosis (LCH) of the female genital tract is very uncommon. Since the natural history of this tumor is unpredictable, with spontaneous remissions and exacerbations, establishment of the diagnosis is often difficult. In the few reported cases of LCH involving the vulva there were systemic manifestations of the disease. We report the clinical and histopathological findings in the case of a 36-year-old woman who presented with a 9-year history of vulvar lesions. The diagnosis of LCH was established by immunohistochemical techniques, which demonstrated sheets of S-100 protein positive histiocytes in the dermal tumor. The patient received radiotherapy to the vulva and responded with complete remission. PMID- 8244197 TI - Primary ovarian leiomyoma. A case report. AB - A 32-year-old woman was admitted for termination of her 6-week pregnancy at her request. However, pelvic examination followed by transvaginal ultrasonography (TVU) revealed an adnexal mass complicating pregnancy and requiring surgical intervention that was carried out after termination of pregnancy. At laparotomy, left salpingo-oophorectomy was performed and histologic examination revealed a leiomyoma arising primarily in the ovary. PMID- 8244198 TI - 12-0-Tetradecanoylphorbol 13-acetate (TPA)-induced enzymatic change of human non T, non-B lymphoid leukemia cell lines. AB - Culture of two lymphoid leukemia cell lines with 5 x 10(-9) to 10(-7) M 12-0 tetradecanoyl-phorbol 13-acetate (TPA) induced the significant increase in sialyltransferase, acid phosphatase and butyrate esterase activities, the decrease in poly(A) polymerase and terminal deoxynucleotidyl transferase (TdT) activities. B4-, J5-, TdT- or PNA-positive cells were decreased in TPA-treated cells, while B1-positive cells were increased. These results suggest enzymatic changes as an aspect of TPA-induced differentiation in lymphoid leukemia cell lines. These enzymatic activities may be useful markers for the differentiation of lymphoid leukemia cells. PMID- 8244199 TI - Bleeding as a cause of death in HIV-seropositive patients with congenital clotting disorders. AB - The causes of death in a group of HIV-seropositive patients suffering from congenital clotting disorders (cCD) were studied. During the past 6 years, we have followed 19 patients with cCD and HIV infection. Eight patients fulfilled revised CDC criteria for AIDS, 6 subjects reached stage III of CDC, and 5 remained asymptomatic (CDC stage II). All patients who developed AIDS died. In 5 patients, the terminal cause of death was a severe haemorrhage (hematemesis, melena or haemoptysis) after gastrointestinal or lung opportunistic infections. Two other patients died as a consequence of disseminated infections but without significant bleeding. Only one subject died due to neoplastic disease. In the first stages of CDC (II and III), no increase in bleeding symptoms were seen in cCD HIV infected patients. The risk of haemorrhages is, however, increased in AIDS patients. Adequate replacement therapy should be started early whenever severe opportunistic infections appear. PMID- 8244200 TI - Platelet function and survival in multiple myeloma. AB - Symptoms of haemorrhagic diathesis are frequently observed in multiple myeloma patients. Their nature is complex and is thought to be connected with platelet function disturbances, coagulation abnormalities and vascular endothelium damage. We demonstrated the shortening of platelet halflife in six myeloma patients. Disturbed thrombocytopoiesis, decreased platelet adhesion and aggregation were shown in all patients studied. The shortened platelet survival in our patients may have resulted from disturbed platelet function resulting from high immunoglobulin concentration and changes in thrombocytopoiesis. PMID- 8244201 TI - Hyperkalaemia with renal tubular dysfunction by sulfamethoxazole-trimethoprim for Pneumocystis carinii pneumonia in patients with lymphoid malignancy. AB - Hyperkalaemia with renal tubular dysfunction by oral therapy of sulfamethoxazole trimethoprim (co-trimoxazole) is described in 2 elderly Japanese patients with lymphoid malignancy, who developed Pneumocystis carinii pneumonia and improved. A high dose of cotrimoxazole induced hyperkalaemia with the elevation of serum creatinine and blood urea, and increased urinary N-acetyl glucosaminase after several days of the drug administration in these patients; one patient became unconscious. Discontinuation of co-trimoxazole normalized serum potassium level and symptoms. A repeated low dose of the drug induced hyperkalaemia. Before the treatment of co-trixomazole, their serum levels of creatinine showed upper limits of normal ranges. In the present study, our cases suggested that patients receiving a high dose of co-trimoxazole should be evaluated for these potential complications during a course of treatment, particularly in elderly patients with preexisting renal dysfunction. PMID- 8244202 TI - Adequate iron stores and the 'Nil nocere' principle. AB - There is a need to change the policy of unselective iron supplementation during periods of life with physiologically increased cell proliferation. Levels of iron stores to be regarded as adequate during infancy and pregnancy are still not well established. Recent data support the view that it is not justified to interfere with physiological adaptations developed through millions of years by sophisticated and precisely coordinated regulation of iron absorption, utilization and storage. Recent data suggest that the chelatable intracellular iron pool regulates the expression of proteins with central importance in cellular iron metabolism (TfR, ferritin, and erythroid 5-aminolevulinic synthetase) in a coordinately controlled way through an iron dependent cytosolic mRNA binding protein, the iron regulating factor (IRF). This factor is simultaneously a sensor and a regulator of iron levels. The reduction of ferritin levels during highly increased cell proliferation is a mirror of the increased density of TfRs. An abundance of data support the vigorous competition for growth essential iron between microbial pathogens and their vertebrate hosts. The highly coordinated regulation of iron metabolism is probably crucial in achieving a balance between the blockade of readily accessible iron to invading organisms and yet providing sufficient iron for the immune system of the host. The most evident adverse clinical effects of excess iron have been observed in immunodeficient patients in tropical countries and in AIDS patients. Excess iron also increases the risk of initiation and promotion of malignant processes by iron binding to DNA and by the iron-catalysed release of free radicals. Oxygen radicals were shown to damage critical biomolecules leading, apart from cancer, to a variety of human disease states, including inflammation and atherosclerosis. They are also involved in processes of aging and thrombosis. Recent clinical trials have suggested that the use of iron-chelators, natural and synthetic antioxidants, and anti-TfR monoclonal antibodies can contribute in retarding malignant cell proliferation. Hypoferraemia during pregnancy is--like haemodilution--an adaptation to the risks involved in the natural hypercoagulable state of pregnancy. It may also serve to prevent the risk of infections and mutagenicity in the highly proliferating tissues of the foetus. Blunted erythropoiesis has been revealed during the first 30 weeks of pregnancy by the use of the newly developed method of determining the soluble serum transferrin receptor. The lack of increase in erythropoietin levels proves that there is no hypoxia. Decreases in Hb and iron levels are parts of a physiological adaptation. As a consequence they should neither be treated nor prevented. It is stressed that whenever a widespread and ingrained routine medical intervention has to be changed it is essential to first monitor the potential health effects of the recommended change in a national policy. PMID- 8244203 TI - Human glucose-6-phosphate dehydrogenase: structure and function of normal and variant enzymes. AB - Glucose-6-phosphate dehydrogenase (G6PD) is a unique enzyme with many genetic variants. Recent progress in molecular biological techniques have provided several important findings about the structure-function relationship of human G6PD. A putative substrate glucose-6-phosphate binding (around Lys 205) and a putative 'structural' NADP binding site (around Lys 386) have been identified. A conservative segment (amino acid 38-44) near the N-terminus was proposed to be the possible second NADP binding region (amino acid 38-44). Analysis of amino acid substitutions of variants revealed that there might be some relationship between the position of substitution and the properties of variants. PMID- 8244204 TI - An alternative view of autoimmunity: the network of antibody forming cells and the immunological homunculus. PMID- 8244205 TI - Dose-dependent effects of hydatid fluid toxins from Echinococcus granulosus on mouse peritoneal macrophages. AB - Hydatid fluid (HF) from hydatid cysts of the cestode Echinococcus granulosus inhibited the phagocytosis of bacteria and yeast cells by host macrophages in vitro. Different assays were used to study the dose-dependent effect of partially purified HF toxins on peritoneal macrophages (PM phi) of the mouse. Trypan blue exclusion, as well as measuring the specific release of lactate dehydrogenase activity and 51Cr release of 51Cr-labelled PM phi, showed that incubation with HF toxins lead to the lysis of the target cells. By measuring the effect of the toxins on the reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide by PM phi, and by experimental labelling using [3H]-uridine, a decrease in metabolic activity was seen. However, at non-lytic concentrations, the PM phi showed a peak of metabolic activity. The observations of the effects on macrophages by parasite-derived toxins may be related to a mechanism by which the parasite survives within an immunized host. PMID- 8244206 TI - Experimental infection in chickens with larvae of Baylisascaris transfuga (Nematoda: Ascaridoidea). AB - The larvae of Baylisascaris transfuga (Rudolphi, 1819) were able to penetrate the liver, lungs, carcass and brain of infected chickens, but a great number of larvae accumulated in the liver. No clinical signs were seen. Birds may serve as paratenic hosts of the parasite, but B. transfuga seems not to be a possible agent of avian cerebrospinal nematodosis. PMID- 8244207 TI - Cross-mating between Amblyomma parvum Aragao, 1908 and Amblyomma pseudoparvum Guglielmone, Mangold et Keirans, 1990 (Acari: Ixodidae). PMID- 8244208 TI - Features of bovine demodecosis (Demodex bovis Stiles, 1892) in Mongolia: preliminary observations. PMID- 8244209 TI - Isolation of a rickettsial organism (Eperythrozoon) from mice during virological testing of ixodid ticks. AB - A strain of Eperythrozoon (Rickettsiales: Anaplasmataceae) was isolated from suckling SPF mice (SM) inoculated with a homogenate of nymphal Ixodes ricinus (L.) ticks. The strain, passaged in the SM brain, was partially filterable through a 220-nm Millipore membrane, moderately sensitive to diethylether, and pathogenic for SM while no symptoms were observed in inoculated adult mice or male guinea pigs. A conspicuous cytopathic effect was produced in PS and SPEV (embryo pig kidney) stable cell lines. Both the cell cultures and SM could be used for the infectivity titration of the agent. PMID- 8244210 TI - Experimental transmission of Caryospora bigenetica Wacha et Christiansen, 1982 (Apicomplexa: Eimeriidae) from a rattlesnake, Crotalus atrox, to rodents and pigs. AB - Caryosporan oocysts were found in the feces of a diamond rattlesnake, Crotalus atrox, kept in ZOO Usti nad Labem, Czech Republic. Comparison of oocyst structure with hitherto described Caryospora species from snakes revealed that they were Caryospora bigenetica Wacha et Christiansen, 1982. The rattlesnake, Crotalus atrox, represents a new host for C. bigenetica. The common vole (Microtus arvalis) and gerbil (Meriones unguiculatus) were successfully infected with a mixture of C. bigenetica oocysts and sporocysts following oral inoculation. All experimentally infected animals displayed clinical signs of dermal coccidiosis, including swellings of the face, ears, footpads, and scrota or vagina. Gerbils developed more severe clinical signs of infection and had a higher mortality than voles. Developmental stages of C. bigenetica were found in connective tissue of the nose, cheeks, ears, scrotum or vagina. Transmission of C. bigenetica caryocysts between common voles and mice, between mice and common voles, common voles and gerbils, and common voles and pigs was demonstrated. This study demonstrates that C. bigenetica can be transmitted by predation or cannibalism between different species of rodents and pigs. PMID- 8244211 TI - [Cytokine and inflammation: role of IL-5 and its receptor system in inflammation]. AB - The inflammatory response is mediated by various cytokines. IL-5 is one of the inflammatory cytokines. The study of IL-5 has its origins in the search for one of the B-cell differentiation factors, named T-cell-replacing factor (TRF), that induces antigen-stimulated B cells to differentiate into antibody-forming cells. Eosinophil-differentiation factor, EDF, is a factor produced by thoracic duct lymphocytes of parasite-infected rats. cDNA cloning and mAb against IL-5 enable us to identify this molecule as a cytokine (TRF/EDF) that has pleiotropic activity on various target cells besides B cells and eosinophils. The pleiotropic activity of IL-5 is directly dependent on the initial binding to the IL-5 receptor (IL-5R) on the target cell surface. IL-5 transduces its signals through high affinity IL-5R which is constructed by two distinct polypeptides, alpha and beta. IL-5R alpha binds IL-5 with low affinity and associates with the beta chain which can convert low affinity IL-5R to high affinity IL-5R. IL-5R beta that does not bind IL-5 by itself is essential for the IL-5 signaling and is shared among IL-5R, IL-3R, and GM-CSFR. These results imply why IL-5, IL-3, and GM-CSF are eosinophylopoietin. The truncated IL-5R alpha and IL-5R beta complexes can not transduce IL-5 signals, although they bound IL-5 with high affinity, suggesting that IL-5-specific signaling may be transduced through IL-5R alpha. PMID- 8244212 TI - [Present and future of the methods for measurement of intracellular Ca2+ concentration]. AB - The development of fluorescent Ca2+ indicators, which have made measurement of the dynamic change in cytosolic free Ca2+ concentration during biological activity feasible, has allowed correlations to be drawn between biochemical changes within cells and their functions. Recently, many new types of Ca2+ indicators with special merit and also new methods for fluorescence measurement have been published. The present technical review summarizes properties of the newly developed indicators and describes the newly developed methods for measuring intracellular Ca2+ concentration. In particular, examples of image analysis applied to single cells and tissue slice preparations are stated in detail. Furthermore, a newly developed single optical fiber device for making measurements in the deep brain region is also introduced. PMID- 8244213 TI - [Effects of quinapril on the development of hypertension, cardiac hypertrophy and stroke in salt-sensitive Dahl rats and stroke-prone spontaneously hypertensive rats]. AB - The effects of repeated administration of quinapril (10 mg/kg/day) on the development of hypertension, cardiac hypertrophy and survival rate were examined, and compared with those of enalapril (10 mg/kg) in salt-sensitive Dahl (Dahl S) rats and 1% saline-loaded stroke-prone spontaneously hypertensive rats (SHRSP). The Dahl S rats were treated with the drugs at 10 to 20 weeks of age and the SHRSP, at 8-19 weeks of age. (1) In the Dahl S rats, salt loading rapidly raised systolic blood pressure, which was around 220 mmHg at 10 weeks of age. Both quinapril and enalapril significantly prevented the development of hypertension (below 160 mmHg) and cardiac hypertrophy. Age-associated histopathological alterations in the kidney and mesenteric artery in Dahl S rats were suppressed by the drug treatment. (2) Salt-loaded SHRSP rapidly developed severe hypertension (270 mmHg at 12 weeks of age) accompanied with stroke signs, and 19 animals out of 20 died by the end of the experiment. Both quinapril and enalapril significantly inhibited the age-associated development of hypertension and markedly improved the survival rate (only two animals out of 16 died in both groups). These results suggest that quinapril has protective actions against age associated development of hypertension and cardiac hypertrophy, and as a result, it prolongs the life span. PMID- 8244214 TI - [The maxillary sixth-year molar and its relation to the maxillary sinus. A comparative study between the panoramic tomogram and the computed tomogram]. AB - Panoramic radiographs (OPGs) and computer tomograms (CTs) of 30 patients, the latter taken on average 2.5 months later, were evaluated for the purpose of comparing the relation of their roots to the bottom of the maxillary sinus. The CT and OPG images yielded significant differences in the topographic relations between the roots and the maxillary sinus. In the OPG 64 out of 129 roots seemed to penetrate the maxillary sinus. In the CT transversal slices, on the other hand, penetration could be observed for only 37 roots. This phenomenon became more apparent when the relations between the mesial, distal, buccal, and oral root surfaces and their relations to the sinus were assessed. Applying a three stage scoring scheme, it was found that in the large majority of cases only the apices were in contact with the mucosa of the sinus. Thus, the possible loss of anchorage in these cases is below five percent. PMID- 8244215 TI - [The effect of information and communication in the orthodontic consultation. 1. The imparting of the information]. AB - The initial consultation has a crucial impact on the patient's decision whether or not to undergo orthodontic treatment. The consultation is the occasion for providing information regarding the nature of the malocclusion and its possible treatment. The consultation has a threefold objective: first, the ethical, i.e. assisting the patient in making an informed and responsible decision; second, the legal, i.e. facilitation of an informed consent; and third, the psychological, i.e. motivation to treatment and enhanced health behavior. The basic prerequisites to a successful consultation are the patient's accurate comprehension and retention of the given information. In the course of an ongoing quasi-experimental study the Wuerzburg orthodontic department has made video recordings of initial consultations. All information was coded according to a standardized schematic. After ten days the patients were interviewed about the consultation. With help of the schematic, retention and comprehension of the information given were assessed quantitatively by comparing the number of information units actually given with those recalled, understood, or forgotten. In addition, the recordings were analyzed for provider interaction patterns. PMID- 8244216 TI - [The orthodontic findings in patients with unilateral Angle Class II]. AB - In this study the original orthodontic documentation of 23 patients (twelve girls and eleven boys, average age 12.7 years) with one side Angle class II subdivision were analyzed. Computer assisted measurements were obtained from dental models and from posteroanterior and lateral oblique cephalometric radiographs of each patient. This results of this research indicate that the asymmetric occlusion was caused by dental as well as by skeletal components. In addition to an asymmetric position of the upper first molars, a lateral position of the whole mandible was established. PMID- 8244217 TI - [The accuracy of holograms compared to other model measurements]. AB - Because holograms are easy to deal with, and because their information content is especially high, they will potentially play a much larger role in the future. In this study plaster casts were placed in a measuring device fabricated in our department, measured by means of a laser beam, and then the accuracy of these measurements was compared to hologram measurements taken by means of a sliding caliper. The result was that the computer-guided hologram measurements evidenced no significant differences with the measurements with a laser beam. The hologram measurements, which are regarded as having an accuracy of 0.182 mm, should therefore be significantly superior to sliding caliper measurements. When measuring the mesiodistal width of teeth, the caliper yields a pseudo-accuracy which doesn't correspond to the factual conditions. When defining the Z coordinates, the sliding caliper's measuring deficiencies are easily circumvented, but this does not automatically imply a higher degree of accuracy. PMID- 8244218 TI - [The effect of static magnetic fields on prostaglandin synthesis in L-929 and 3T3 mouse fibroblasts. An in-vitro study]. AB - The aim of this study was to examine the influence of static, inhomogeneous magnetic fields on prostaglandin synthesis in L-929 and 3T3 mouse fibroblasts. Two pairs of magnets, one CoSm and the other NdFeB, were attached 10 mm apart under the culture chamber. One pair was in the attracting position and the other in the repelling position. The maximum magnetic induction measured was 85 respectively 120 mT and the incubation time was 72 hours. Control groups consisted of untreated cells and cells under the influence of demagnetized magnets. The following measurements were taken: prostaglandin synthesis (PGE2), protein content (GP), and the number (Z) of cells (only L-929 mouse fibroblasts). The data were analyzed statistically using the Tukey test and the significance niveau was set at alpha = 0.05. The results using this method demonstrated that the measurable effects of in vitro treatment of mouse fibroblasts with static magnetic fields of varying induction did not significantly differ from the control groups. The sensitivity of our method was verified beforehand with a recombinant, human TNF alpha stimulated reference group. PMID- 8244220 TI - Joint Meeting of the Deutsche Gesellschaft fur Pharmakologie und Toxikologie and of the Association Francaise des Pharmacologistes. Lille, France, 6-8 October 1993. Abstracts. PMID- 8244219 TI - [After myocardial infarction, captopril improves the prognosis]. PMID- 8244221 TI - Health care under French national health insurance. AB - Several elements of the French health system--the predominance of office-based medical practice, the mix of private and public hospitals, the use of patient cost sharing, direct payment of physicians by patients, and financing derived from payroll taxes--closely resemble aspects of the U.S. health system. There are four major differences between the two systems: the French system covers more than 99 percent of the population; the prices of health services in French are lower than in the United States; the volume of most services is higher than in the United States; and French health care spending per capita is lower than in the United States. Recently enacted and proposed reforms in France likely will strengthen existing health spending targets and utilization controls. PMID- 8244222 TI - Budget limits and managed competition: allies, not antagonists. PMID- 8244223 TI - Antitrust enforcement: putting the consumer first. PMID- 8244224 TI - Antitrust policy and real health care reform. PMID- 8244225 TI - Physicians as professionals, not pawns. PMID- 8244226 TI - Pharmaceutical companies as a positive force for change. PMID- 8244227 TI - Flexibility in antitrust enforcement. PMID- 8244228 TI - U.S. health care costs: the untold true story. PMID- 8244229 TI - Hospital/physician relationships: a constraint to health reform. PMID- 8244230 TI - Physician incomes under an all-payer fee schedule. AB - This DataWatch estimates physicians' net incomes under various scenarios for universal adoption of the Medicare fee schedule. American Medical Association (AMA) data on physicians' gross incomes by payer are adjusted for differences in fees across Medicare, Medicaid, and private payers. Net income (gross income net of practice costs) is shown under several alternatives. Average net incomes would range from roughly $100,000 to $200,000, depending on specialty and the particular assumptions used. General practitioners' incomes would typically rise, and income differences across specialties would be narrowed significantly. The Medicare fee schedule, as intended, would increase incomes of primary care physicians relative to specialists. PMID- 8244231 TI - Explaining the recent growth in Medicaid spending. AB - Medicaid spending more than doubled from 1988 to 1992, reversing a long trend of cost containment in the program. Reasons for the cost explosion are severalfold. (1) Congress expanded eligibility to more children, pregnant women, and low income elderly persons. (2) The recession has added more people to the Medicaid rolls. (3) Growing numbers of disabled cash assistance recipients have increased Medicaid enrollment. (4) States have increased their use of federal Medicaid funds to supplement previously state-funded programs and have become more skillful in leveraging federal funds to defray the cost of their Medicaid programs. This DataWatch explores the relative impact of enrollment changes, inflation, and increased reimbursement on the increase in Medicaid spending. PMID- 8244232 TI - Health reform lessons learned from physicians in three nations. AB - To explore the concerns of practicing physicians as a way to inform the health reform debate, the authors conducted a survey of physicians in the United States, Canada, and Germany. Survey results indicate that U.S. physicians are most likely to view affordability as the greatest barrier to access to care for their patients. However, unavailability of services and long waiting times were cited most often by Canadian physicians. German physicians did not cite access problems as frequently as Canadian physicians did; other measures of satisfaction were closer to U.S. levels, suggesting fewer trade-offs if the United States were to adopt aspects of the German health care system. PMID- 8244233 TI - How well do Americans understand their health coverage? AB - Data from two surveys are used in this DataWatch to explore Americans' understanding of their health insurance. First, data from a national survey of consumers are used to examine if people with private health insurance correctly report their coverage for six services. Second, information from an evaluation of a pilot project of subsidized insurance in New York is used to investigate how well newly insured persons understand their coverage. Based on these surveys, almost all privately insured people understand the basic elements of their insurance plans but underestimate their coverage for mental health, substance abuse, and prescription drug benefits and overestimate their coverage for long term care. People who are newly insured in physician networks or health maintenance organizations seem uncertain about what services their plan covers and restrictions on their choice of hospitals. PMID- 8244234 TI - Trends in hospital/physician relationships. AB - Findings from two surveys of all physicians in Pima County, Arizona, in 1985 and 1990 to determine the trajectory of hospital/physician relationships suggest that conflicts have increased in almost all areas. Physicians report the greatest number of and increase in conflicts in traditional problem areas such as the quality of nursing and response to equipment requests. They report a smaller increase in conflicts relating to the new competitive marketplace, including hospital efforts to influence medical practice. Both types of problems are most prevalent among younger physicians. The new marketplace thus appears to exacerbate traditional hospital/physician conflicts more than it generates new areas of contention. PMID- 8244235 TI - Access to care in the Indian Health Service. AB - The Indian Health Service (IHS) is unique among U.S. private and public health programs in that free comprehensive health services are provided to eligible American Indians and Alaska Natives regardless of their ability to pay. However, resource limitations may compel some eligible persons to go outside of the IHS system to receive health care. Although IHS eligibles have comparatively low rates of private or public health care coverage, and much of this population lives in underserved areas, over half of IHS-eligible persons had some type of out-of-plan use in 1987. Furthermore, services received through private providers appear to supplement those received through IHS-sponsored providers. Overall, persons who use both IHS and non-IHS providers have higher levels of health care use than do those who rely exclusively on the IHS. PMID- 8244236 TI - The financial capacity of nonprofit hospitals. PMID- 8244237 TI - Boston teaching hospitals' response to the Kane Report. PMID- 8244238 TI - Opportunities in mental health services research. PMID- 8244239 TI - AIDS. PMID- 8244240 TI - Why managed care has failed to contain health costs. AB - Much evidence points to the fact that managed care plans (health maintenance organizations and preferred provider insurance) reduce costs and offer value for money. Yet they apparently have not helped to slow national health expenditures. One explanation is that the practices of purchasers (including government and employers), the tax laws, and other market imperfections have reduced the demand for real cost containment, depriving managed care plans of an adequate incentive to cut cost and price. These market conditions can and should be corrected; the managed competition proposal being discussed at the national level is a comprehensive plan for doing so. PMID- 8244241 TI - Hawaii's message to the nation. PMID- 8244242 TI - AMCs: committed to managed care. PMID- 8244243 TI - Understanding faculty practice revenue. PMID- 8244244 TI - Why managed competition cannot be enacted. PMID- 8244245 TI - Markets, budgets, and health care cost control. AB - Health care cost control is often debated in terms of markets versus bureaucracies. Market restraints are limited in practice by the goal of providing access to care. Therefore, effective cost control requires budgeting. Experience from budgeting for other services should put health care cost control in perspective: The goal should not be rational and efficient allocations, merely better ones. The important choice is not between markets and bureaucracy, but rather which decisions should be made by physicians and which by budgeters, and how to ensure that professional judgment is applied where most important. PMID- 8244246 TI - The nation's health care enterprise. PMID- 8244247 TI - Health care reform and rural health networks. AB - Health care reform is likely to raise unique issues for rural communities and providers. This paper identifies and discusses several of these issues, with a particular focus on the potential relationship between health care reform and rural health networks. Topics addressed include the likely impact of health reform on the organization and development of rural health networks, the reimbursement of rural providers, rural medical practice, and state roles in the organization, delivery, and oversight of rural health care. PMID- 8244248 TI - Health spending by state: new estimates for policy making. AB - A new data set from the Health Care Financing Administration gives estimates of state spending for hospital care, physician services, and retail purchases of prescription drugs, which together account for 70 percent of health expenditures nationwide. Analysis of these data, which are the first uniform state data to be produced for nearly ten years, shows considerable variation among states and among regions in health care spending. The New England and Mideast regions show the consistently highest spending levels for all three categories; the Southwest and Rocky Mountain regions spent the smallest amount (as much as 17 percent below the U.S. average). PMID- 8244249 TI - Health care reform in rural areas. PMID- 8244250 TI - Medicaid and health reform: the case of New York. AB - New York State has the largest, most expensive state Medicaid program in the country. Thus, an examination of its Medicaid program can offer valuable lessons for other states that are considering reform of their health systems, as well as for reform at a nationwide level. Much recent growth in Medicaid in New York stems from shifting state-funded human service programs onto Medicaid and shifting the state's share of Medicaid onto nontraditional revenue sources. In contrast to other states, in which Medicaid is an unpopular program, New York's Medicaid provider constituency is large and diverse, and its clientele is relatively white and middle class. These two constituencies have made Medicaid harder to cut than in other states, in which Medicaid recipients lack political and economic clout. Current versions of national health reform will have little effect on Medicaid spending in New York, since they address neither spending on the elderly nor the "Medicaiding" of programs and revenue sources. PMID- 8244251 TI - Recent growth of Medicare home health. AB - Medicare spending for home health services has grown at an unprecedented rate since regulations were revised in 1989. To address the implications of this striking growth, it is necessary to examine the causes. This paper finds that the most important source of growth has been in the number of home health visits received per beneficiary, not in the number of beneficiaries that have been added in response to redefined eligibility criteria. Policymakers should clarify the objectives of the Medicare home health benefit, in light of increased attention to long-term care financing and expanded services for older Americans. PMID- 8244252 TI - Rapid sequence variation of the hypervariable region of hepatitis C virus during the course of chronic infection. AB - To evaluate the clinical significance of sequence variations in the hypervariable region of hepatitis C virus during the course of chronic infection, we performed pairwise comparison of the predominant nucleotide sequences. Hepatitis C virus RNA was extracted from two plasma samples obtained from 12 chronically infected Japanese patients over approximately 1 yr. Complementary DNA containing the hypervariable region was amplified by means of reverse transcription-polymerase chain reaction and was directly sequenced for determination of predominant sequences. In all 12 individuals, the predominant sequence of the hypervariable region at the second time point differed from that at the first time point, and significant sequence variation was observed during this short period. A high proportion of these nucleotide substitutions (90%) resulted in changes of predicted amino acid sequences, indirect evidence of in vivo positive selection for these variants. There appeared to be an important difference in the rate of nucleotide sequence variation of the hypervariable region between four patients with flare-ups of their ALT levels (1.54 to 2.24 x 10(-1)/genome site/yr) and eight patients with quiescent courses (0.13 to 1.21 x 10(-1)/genome site/yr). These results demonstrate that rapid sequence variations of the hypervariable region of predominant virus population take place during the natural course of chronic hepatitis C virus infection. These sequence variations seem to occur as an adaptive response of hepatitis C virus to evade host immunity and may play a major role in the establishment of persistent infection and in the occasional flare-up of hepatitis. PMID- 8244253 TI - Monitoring of antiviral therapy with quantitative evaluation of HBeAg: a comparison with HBV DNA testing. AB - The serological endpoint of response in the treatment of chronic hepatitis B is the loss of hepatitis B virus DNA and HBeAg. Because the quantitative measurement of hepatitis B virus DNA in serum has been shown to be useful for monitoring and predicting response to interferon-alpha therapy, we decided to evaluate whether changes in HBeAg concentration could also be used in this manner. Twenty-nine patients who were initially positive for HBeAg and HBV DNA were serially evaluated for HBeAg concentration with a microparticle-capture enzyme immunoassay. HBeAg levels in serum were calculated by means of comparison with a standard curve of fluorescence rate vs. HBeAg concentration. The results, expressed in milliunits per milliliter, were compared with hepatitis B virus DNA levels determined by means of solution hybridization. The baseline HBeAg concentration proved to be the best independent predictor of response on stepwise Cox regression analysis (p = 0.026). Similar disappearance curves were observed for the two markers, although hepatitis B virus DNA became undetectable at an earlier interval in 13 of 16 cases (81%). In the 16 responders, a decline in HBeAg concentration of more than 90% was observed by wk 12 of therapy (mean +/- S.D., 95% +/- 13%). Nonresponders did not demonstrate such steep declines in HBeAg values by wk 12 (mean +/- S.D., 45% +/- 27%), and levels tended to increase at subsequent time points. We conclude that serial monitoring of HBeAg concentration with a technique that should be readily adaptable to clinical laboratories may be useful in the initial evaluation and monitoring of patients undergoing antiviral therapy. PMID- 8244254 TI - Persistence of hepatitis B virus DNA in the liver after loss of HBsAg in chronic hepatitis B. AB - To determine whether patients with chronic hepatitis B who lose hepatitis B virus DNA and HBsAg from the serum completely resolve the hepatitis and virus infection, we evaluated serum and liver tissue from 11 patients who had lost HBsAg. These patients were evaluated for clinical, histological and serological features and for hepatitis B virus DNA by use of hybridization and polymerase chain reaction techniques. Liver biochemical test results were normal in all except two patients who had mild aminotransferase elevations. All sera were negative for hepatitis B virus DNA by direct hybridization, and only one was positive transiently by polymerase chain reaction. Liver histology was abnormal in all patients, but the changes were mild and markedly improved compared with biopsy specimens taken before loss of HBsAg. Liver tissue from 10 patients was positive for hepatitis B virus DNA by polymerase chain reaction but not by direct hybridization. These findings indicate that loss of HBsAg is associated with marked improvements in clinical and serum biochemical features of chronic hepatitis B, yet mild degrees of hepatitis and low levels of viral DNA may persist in the liver. PMID- 8244255 TI - Quantitation and typing of serum hepatitis C virus RNA in patients with chronic hepatitis C treated with interferon-beta. AB - We quantified serum hepatitis C virus RNA titers and determined hepatitis C virus subtypes in chronic hepatitis C patients treated with interferon-beta to investigate relationships among serum ALT response, serum hepatitis C virus titer and hepatitis C virus subtype. Of 146 chronic hepatitis C patients who received interferon-beta therapy, 24 patients with sustained serum ALT normalization (complete responders) and 26 patients without serum ALT normalization (nonresponders) were randomly selected. Detection, typing and quantitation of hepatitis C virus were performed by means of the "single-tube" polymerase chain reaction method. Of the 24 complete responders, 21 (87.5%) became negative for hepatitis C virus RNA, whereas 21 (80.8%) of the 26 nonresponders remained positive. Hepatitis C virus infections with types I, II, III, IV, II + III and III + IV occurred in 0 (0%), 22 (51.2%), 10 (23.3%), 1 (2.3%), 7 (16.5%) and 3 (7.9%) patients, respectively. The mean pretreatment hepatitis C virus RNA titer of complete responders (0.4 +/- 2.0 x 10(4) CID50/ml) was significantly lower than that of nonresponders (3.8 +/- 4.5 x 10(4) CID50/ml) (p < 0.01). Regardless of HCV subtype, patients with more than 10(4) CID50/ml of HCV did not show serum ALT normalization, whereas complete serum ALT response was seen in most cases with less than 10(2) CID50/ml HCV. These results show that mixed infections with different hepatitis C virus subtypes appear to be more common than previously reported and that the pretreatment serum level of hepatitis C virus RNA is a more important predictor of outcome of interferon therapy than is virus genotype. PMID- 8244256 TI - Viral pathogenesis of hepatocellular carcinoma in the United States. AB - Chronic hepatitis B virus infection is closely associated with the development of hepatocellular carcinoma, which is a major cause of cancer death worldwide. Recent studies have implicated hepatitis C virus infection as a major pathogenic agent of HBsAg-negative hepatocellular carcinoma. The significance of hepatitis C virus and hepatitis B virus infections in the occurrence of HBsAg-negative hepatocellular carcinoma has not been well established in the United States. We studied 91 HBsAg-negative American patients with hepatocellular carcinoma for evidence of hepatitis C virus or hepatitis B virus infection. These patients had no other predisposing factors to hepatocellular carcinoma. A sensitive polymerase chain reaction was employed to detect hepatitis C virus RNA and hepatitis B virus DNA in serum and liver. Three sets of hepatitis C virus and hepatitis B virus primers were used to optimize the detection of viral genomes. Hepatitis C virus antibodies were measured with second-generation immunoassays. Twenty-six (29%) of these patients carried low levels of hepatitis B virus DNA in either serum, liver/tumor tissue or both. On the basis of the results from serological and polymerase chain reaction analyses of serum and liver, we found that 53 of 91 patients (58%) exhibited evidence of hepatitis C virus infection. When data were combined, 14 patients (15%) had evidence of hepatitis B virus/hepatitis C virus coinfection, whereas 12 (13%) were infected with hepatitis B virus alone and 39 (43%) had hepatitis C virus only. Twenty-six (29%) had no markers of hepatitis B virus or hepatitis C virus infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244257 TI - Human leukocyte antigen A1-B8-DR3-DQ2-DPB1*0401 extended haplotype in autoimmune hepatitis. AB - Genetic susceptibility to autoimmune hepatitis is associated with the human leukocyte antigen haplotype A1-B8-DR3 and DR4. To date, only one study in Japan has considered the human leukocyte antigen DP locus in this disease, and no studies have been reported in whites. In this study we used a series of sequence specific oligonucleotide probes to determine human leukocyte antigen DPB1 genotypes in 101 unrelated white northern European patients and 105 racially and geographically matched controls. The aims of the study were twofold: first, to determine the degree of DPB-encoded susceptibility to autoimmune hepatitis, and, second, to establish whether susceptibility can be extended to include human leukocyte antigen DPB. None of 17 DPB1 alleles was significantly associated with the susceptibility to autoimmune hepatitis. Although one particular seven-locus haplotype A1-B8-DRB3*0101-DRB1*0301-DQA1*0501-DQB1*0201-++ +DPB1*0401 was significantly associated with the disease (27% vs. 7%, relative risk = 5.14, p < 0.0005), the association with this haplotype was weaker than that for the six locus haplotype excluding DPB (40% vs. 11%, RR = 5.52, p < 0.0005). When the patients first seen at ages younger than 16 yr (pediatric patients) were considered separately, the greatest relative risk was for the seven-locus haplotype (41% vs. 7%; relative risk = 9.60, p < 0.0005). The results of this study further confirm that major histocompatibility complex-encoded susceptibility to autoimmune hepatitis is located at or close to the human leukocyte antigen DR locus; however, the A1-B8-DR3-DQ2-DPB1*0401 extended haplotype may be important in determining the age of onset and severity of disease. PMID- 8244258 TI - The pathology of hepatitis C as a function of mode of transmission: blood transfusion vs. intravenous drug use. AB - We reviewed the clinical records of 140 consecutively evaluated patients with chronic hepatitis C infection. One hundred twenty-four patients (89%) contracted infection through blood transfusion or intravenous drug use. The liver biopsy specimens of 83 patients (43 blood transfusion cases and 40 intravenous drug abuse cases) were examined without knowledge of the mode of disease transmission. The mean histological activity index score was significantly higher in the blood transfusion group (10.2 +/- 4.2) than in the intravenous drug use group (6.9 +/- 4.5) (p = 0.001). The transfusion group had more periportal bridging necrosis (p = 0.0015) and fibrosis (p = 0.0016) than did the intravenous drug use group, whereas significant differences between lobular degeneration and portal inflammation were not achieved across the two groups. The distribution of final biopsy interpretations also differed significantly between the two groups (p < 0.001), with chronic active hepatitis more frequent in the transfusion group. Moreover, lymphoid aggregates and bile duct damage were more common in patients with chronic hepatitis due to blood transfusion. Multivariate analysis showed that the mode of viral transmission was the most powerful predictor of histological activity index score when tested against patient gender, duration of disease or age at biopsy. One year after completion of this study, 9 of 70 transfused patients and 1 of 54 intravenous drug users had died of liver disease or are awaiting liver transplantation at this writing (p = 0.03). We conclude that transfusion-acquired hepatitis C is associated with more aggressive histological inflammatory activity than hepatitis resulting from intravenous drug use.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244259 TI - Interferon-alpha 2b therapy reduces liver fibrosis in chronic non-A, non-B hepatitis: a quantitative histological evaluation. AB - The aim of this study was to evaluate the effect of interferon-alpha on liver fibrosis with an established quantitative histochemical method for determining collagen as a marker. 59 patients (31 men, 28 women; 47 +/- 14 yr) with chronic non-A, non-B hepatitis (92% with hepatitis C virus antibody) received subcutaneous injections of 3 or 1 MU recombinant interferon-alpha 2b or placebo thrice weekly for 24 wk. Needle-biopsy sections taken before and after interferon treatment were examined for histological evaluation and collagen quantitation. Values were compared with results obtained by means of morphometrical analysis of liver collagen and Knodell scoring histological index. The index of periportal and/or bridging necrosis was the only component of Knodell's histological score significantly decreased (p < 0.05) in patients treated with 3 MU interferon compared with placebo-treated controls. The fibrosis score was not significantly changed. In contrast, liver total collagen variations measured colorimetrically and morphometrically were significantly decreased in patients treated with 3 MU and 1 MU compared with the increase observed in the placebo-treated controls (p < 0.05). From these results, we conclude that a 6-mo course of 3 MU or 1 MU interferon-alpha 2b causes slight but nonetheless significant regression of liver fibrosis as assessed on the basis of quantitative estimation of liver collagen, irrespective of other response criteria, whereas progression of liver fibrosis can be observed in the absence of treatment. PMID- 8244260 TI - Activation of the fibrinolytic system in patients with fulminant liver failure. AB - Abnormalities of blood coagulation and fibrinolysis are a major part of fulminant liver failure. In this study, the key components of the fibrinolytic system were determined in 42 patients with this condition. Admission levels of plasma plasminogen activity were low (9.1% of normal), as to a lesser extent were the activities of its inhibitors alpha 2-antiplasmin (20.5% of normal) and C1 inhibitor (64% of normal). Tissue plasminogen activator activity was found at normal levels, whereas the level of its fast inhibitor, plasminogen activator inhibitor-1, was greatly increased compared with that of controls (24.3 U/ml and 7.4 U/ml, respectively), indicating a shift toward inhibition of fibrinolysis in these patients. Thrombin-antithrombin complex levels in plasma were significantly increased in fulminant liver failure compared with those in controls (33.5 micrograms/L vs. 2.5 micrograms/L, p < 0.001), indicating activation of coagulation in these patients. High plasma levels of D-dimer, a fragment of cross linked fibrin, were also found (1,510 micrograms/L vs. 33 micrograms/L in controls, p < 0.001); this finding correlated with the increased level of thrombin-antithrombin complex (r = 0.61, p < 0.001), consistent with activation of fibrinolysis resulting from intravascular coagulation. In conclusion, there are gross abnormalities of the fibrinolytic system in fulminant liver failure, but because inhibitory activity appears to be present in adequate quantities, this limits the incidence of bleeding due to fibrinolysis. PMID- 8244261 TI - The role of transjugular liver biopsy in fulminant liver failure: relation to other prognostic indicators. AB - Early and accurate diagnosis and prognosis of patients with fulminant liver failure is of critical importance for optimum management. We investigated the role of transjugular liver biopsy in the management of patients with fulminant liver failure and assessed its value in comparison with the recently proposed King's College criteria. Sixty-one patients with fulminant liver failure, ages 2 to 82 yr, were retrospectively analyzed. The main outcome measures were survival vs. death or progression to orthotopic liver transplantation. Transjugular liver biopsy was successful in 60 of 61 patients, with a mean core tissue length of 2.1 cm. There were eight minor complications, all of which were managed conservatively. Biopsy specimens were evaluated for degree of fibrosis, percentage of hepatocellular necrosis and presence of bile duct proliferation, hepatocellular mitotic figures and binucleate hepatocytes for each of the 54 specimens available for analysis. In 34 of 54 patients (63%), the presumed clinical diagnosis was confirmed by transjugular liver biopsy. In 11 patients the procedure served to clarify clinical uncertainty, whereas in 9 of 54 (16.7%) the diagnosis was altered after transjugular liver biopsy. The percentage of necrosis was the only histological parameter that appeared to have significant discriminatory prognostic value, with only 2 of 19 survivors having greater than 70% necrosis. Twenty-one of these biopsy specimens were reviewed by two pathologists, and their degree of correlation for the various features was assessed. Almost perfect concordance was found between the two pathologists on the percentage of hepatocellular necrosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244262 TI - High plasma concentration of myeloperoxidase in cirrhosis: a possible marker of hypersplenism. AB - Plasma myeloperoxidase levels in patients with cirrhosis were compared with those in patients with chronic hepatitis and healthy controls by means of a specific radioimmunoassay for myeloperoxidase. The mean concentration of plasma myeloperoxidase in cirrhotic patients (309.1 +/- 17.2 ng/ml, n = 41) was markedly higher than that in chronic hepatitis patients (222.6 +/- 17.2 ng/ml, n = 21) (p < 0.01) and normal controls (219.5 +/- 5.7 ng/ml, n = 50) (p < 0.01). Plasma myeloperoxidase showed good negative correlations with neutrocyte count (r = 0.32, p < 0.01), thrombocyte count (r = -0.40, p < 0.01), red blood cell count (r = -0.32, p < 0.01), serum albumin (r = -0.35, p < 0.01), and cholinesterase (r = 0.32, p < 0.02) and positive correlations with serum alkaline phosphatase (r = 0.49; p < 0.01) and lactate dehydrogenase (r = 0.31, p < 0.01) in patients with cirrhosis or chronic hepatitis. Among lactate dehydrogenase isozymes, a good positive correlation was seen between plasma myeloperoxidase and lactate dehydrogenase-2 (r = 0.40, p < 0.01) and lactate dehydrogenase-1 (r = 0.03, p < 0.02). Plasma myeloperoxidase was significantly higher in the cirrhotic and chronic hepatitis patients with splenomegaly (341.1 +/- 19.4 ng/ml, n = 31) than in those without splenomegaly (217.4 +/- 12.2 ng/ml, n = 29) (p < 0.01). We also examined the difference between plasma levels of myeloperoxidase in the portal and peripheral blood.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244263 TI - Histological evidence for recurrence of primary biliary cirrhosis after liver transplantation. AB - Whether primary biliary cirrhosis recurs after orthotopic liver transplantation remains a controversial issue. Sixty consecutive patients with primary biliary cirrhosis with at least 1 yr of follow-up after liver transplantation were studied. All patients were treated with triple-drug immunosuppression (cyclosporine, prednisone, azathioprine). Hepatic biochemical parameters and protocol liver biopsy specimens were evaluated 1 wk, 3 wk, 4 mo and yearly after orthotopic liver transplantation and at times of liver dysfunction. Antimitochondrial antibody titers and IgM levels were determined at 4 mo and yearly. At the time of last follow-up, all patients had marked symptomatic improvement compared with their pretransplant condition, and 91% of the patients had normal hepatic biochemical parameters, including serum levels of alkaline phosphatase, bilirubin, ALT and IgM. In addition, all patients had significant decreases in antimitochondrial antibody titer (p = 0.0001) and significant decreases in serum levels of IgM (p = 0.0001). Forty-one of the 60 patients had near-normal liver histological appearance. Of those with abnormal histological appearance, five patients, 2 to 6 yr after orthotopic liver transplantation, had histological features typical of a florid duct lesion, suggesting recurrent primary biliary cirrhosis. All five patients with portal granulomas had normal hepatic biochemical values and were clinically asymptomatic. Two of the five patients had persistent antimitochondrial antibody titers. We consider the documented histological changes highly suggestive of recurrence of primary biliary cirrhosis after liver transplantation but, so far, have no evidence that the condition in these patients is progressive. PMID- 8244264 TI - A prospective study of biliary cytology in 100 patients with bile duct strictures. AB - In patients with obstructive jaundice due to biliary tract stricture a tissue diagnosis is essential because of the varied treatment options available. Radiological imaging of a biliary stricture may suggest that it is malignant, but only a tissue diagnosis can be conclusive. The difficulty of obtaining biopsy tissue has encouraged the use of cytology in this field. This study prospectively analyzed the diagnostic value of exfoliative bile and brush cytology methods. One hundred consecutive patients with biliary strictures diagnosed at endoscopic retrograde cholangiopancreatography or percutaneous transhepatic cholangiography (60 men and 40 women; median age = 71 yr, range = 31 to 91 yr) underwent biliary cytology and were divided into two groups. Group 1 comprised the first 47 patients, who were studied by means of bile cytology alone; and group 2 comprised the subsequent 46 patients, who were studied by means of bile and brush cytology techniques. Seven patients were excluded from analysis because of inadequate follow-up information. A single experienced cytologist examined all samples to determine whether they were neoplastic. Eighty-one patients had malignant strictures and 12 had benign strictures. Combined bile and brush cytology (group 2) was more sensitive than bile cytology alone (group 1) (69% [27 of 39] vs. 33% [16 of 42], p < 0.01). In the patients studied by means of bile and brush cytology methods (group 2), cytologic study of brushings was more sensitive (69% vs. 26%, p < 0.01). No false-positive results were reported in either group (specificity = 100%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244265 TI - Evidence for neurological dysfunction in end-stage protoporphyric liver disease. AB - Protoporphyria is a genetic disorder characterized by a defect in the enzyme ferrochelatase, which catalyzes the chelation of iron to protoporphyrin. This causes excessive accumulation and excretion of protoporphyrin. The predominant clinical feature is photosensitivity. Progressive and fatal liver disease occurs in a small percentage of cases. We report our experience with eight patients with end-stage protoporphyric liver disease in whom a syndrome developed before transplantation that resembled the neurological crises of the acute porphyrias. This syndrome was characterized by abdominal pain, hypertension, tachycardia, extremity pain and weakness, constipation and nausea and vomiting. Erythrocyte and serum protoporphyrin levels were markedly increased in all patients. In one patient, profound hemolysis developed during the anhepatic phase of transplantation and continued over a period of 72 hr, causing an extreme increase in the serum protoporphyrin level. Progressive weakness deteriorated to paralysis in this patient. This phenomenon suggests that protoporphyrin may gain access to neural tissue when serum levels are markedly increased, causing neurotoxicity. PMID- 8244266 TI - Differential control by arterial and portal noradrenaline of hepatic carbohydrate metabolism: evidence for an indirect hemodynamic mechanism. AB - In an earlier study it was found (a) that arterial noradrenaline caused increases in glucose and lactate output slower in onset, smaller in peak height and clearly longer in duration than those caused by portal noradrenaline and (b) that arterial noradrenaline was extracted to a much greater extent than portal noradrenaline. It is possible that the differences in the actions of arterial and portal noradrenaline were due to the different hemodynamic changes; arterial noradrenaline reduced hepatic flow more strongly and with different kinetics than did portal noradrenaline. Therefore the effects of arterial and portal noradrenaline were investigated in isolated rat liver, single-pass perfused through both the hepatic artery (100 cm H2O, 30% flow) and the portal vein (15 cm H2O, 70% flow), after inhibition of the hemodynamic alterations by the Ca2+ antagonist nifedipine. We found (a) that nifedipine inhibited the hemodynamic changes by arterial noradrenaline strongly and those by portal noradrenaline slightly, (b) that the remaining hemodynamic alterations in the presence of high concentrations of the Ca2+ antagonist (100 mumol/L)--slight decrease in portal flow with unaffected arterial flow--were essentially identical after arterial and portal application of noradrenaline, (c) that the pronounced kinetic differences in the alterations by arterial and portal noradrenaline of glucose and lactate balance were diminished by 50 mumol/L nifedipine and abolished by 100 mumol/L of the Ca2+ antagonist and (d) that the different hepatic extractions of arterial and portal noradrenaline were greatly reduced and almost equalized by nifedipine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244267 TI - Effects of nitric oxide and cyclooxygenase inhibition on splanchnic hemodynamics in portal hypertension. AB - Portal hypertension is associated with splanchnic hyperemia and increased plasma levels of prostacyclin. Recently, nitric oxide was proposed as a mediator of this arterial vasodilatation. We hypothesized that portal hypertension alters the relative contribution of prostacyclin and nitric oxide to splanchnic vasomotor control. We studied the relationship of nitric oxide and prostaglandins in normal and portal-hypertensive (3 wk after partial portal vein ligation) male rabbits at baseline and following increasing doses of indomethacin, LG-nitro-L-arginine methylester or both. L-arginine was used as the control. Aortic, central and portal venous pressures were measured directly. Blood flow in the superior mesenteric artery was measured by means of an ultrasonic flow probe, and resistance was calculated. LG-nitro-L-arginine methylester produced vasoconstriction (increased resistance and decreased blood flow in the superior mesenteric artery) in normal and portal-hypertensive rabbits, although in portal hypertensive animals resistance and superior mesenteric artery blood flow remained significantly different than that in normal rabbits because of preexisting hyperemia. L-arginine reversed the effect of LG-nitro-L-arginine methylester. Cyclooxygenase blockade induced dose-dependent vasoconstriction in normal and portal-hypertensive animals. Indomethacin induced further vasoconstriction after LG-nitro-L-arginine methylester and reduced portal venous pressure in portal-hypertensive animals. We conclude that this indicates an amplified role for some prostaglandin, probably prostacyclin, in portal hypertension hemodynamics. It also implies that the two vasodilators act by way of independent mechanisms. PMID- 8244268 TI - Transforming growth factor-alpha expression during liver regeneration after partial hepatectomy and toxic injury, and potential interactions between transforming growth factor-alpha and hepatocyte growth factor. AB - Transforming growth factor-alpha and hepatocyte growth factor are important stimulators of hepatocyte proliferation. In this series of experiments we sought to measure the expression of transforming growth factor-alpha mRNA by hepatocytes in response to toxic liver injury produced by carbon tetrachloride or galactosamine and to perform a more detailed analysis of transforming growth factor-alpha expression after partial hepatectomy. We also explored the interactions of transforming growth factor-alpha and hepatocyte growth factor in their effects on hepatocytes in vitro and tested the ability of these factors to stimulate endogenous transforming growth factor-alpha production by hepatocytes. In previous work we have used oligonucleotide probes to measure transforming growth factor-alpha mRNA expression after partial hepatectomy. In this study we used a rat transforming growth factor-alpha cDNA probe and found that the level of liver transforming growth factor-alpha mRNA increases 4 hr after partial hepatectomy, shows peak expression at 18 hr and returns to the normal level by 36 to 48 hr. Measurement of the corresponding peptide in the liver by means of radioimmunoassay shows that the level of transforming growth factor-alpha rises by 12 hr, peaks at 24 hr and remains significantly increased at 48 hr compared with the levels in sham-operated rats. Carbon tetrachloride and galactosamine are known to produce different patterns of acute liver injury, with maximal hepatocyte DNA synthesis at 48 hr and 5 days, respectively. After carbon tetrachloride administration the profiles of the transforming growth factor-alpha and hepatocyte growth factor mRNA expression are similar, each showing two peaks: the first at 12 hr and the second at 48 hr. In contrast, after galactosamine induced liver injury the expression patterns of transforming growth factor-alpha and hepatocyte growth factor mRNAs differ: hepatocyte growth factor shows a major peak at 24 hr, with a smaller increase at 5 days, whereas transforming growth factor-alpha begins to increase after 2 days, with a single peak occurring at 5 days. In primary hepatocyte cultures, transforming growth factor-alpha and hepatocyte growth factor appear to have complementary effects. The maximal hepatocyte nuclear labeling index induced by hepatocyte growth factor was 42%; the addition of transforming growth factor-alpha increased this to 74%. Exogenous transforming growth factor-alpha, but not hepatocyte growth factor, stimulates the production of the transforming growth factor-alpha peptide by hepatocytes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8244269 TI - Handling of epidermal growth factor and number of epidermal growth factor receptors are changed in aged male rats. AB - In the aged liver, cell proliferation and induction of DNA synthesis by epidermal growth factor is impaired. Changes in the hepatic handling of epidermal growth factor may be important for these effects. I compared epidermal growth factor handling in the livers of young and old rats. Epidermal growth factor binding capacity of plasma membranes was reduced from 1.30 +/- 0.15 to 0.51 +/- 0.19 pmol/mg in young and old animals, respectively. Intracellular handling of epidermal growth factor was assessed by means of portal injection of 125I-labeled epidermal growth factor and collection of bile samples. The extraction of epidermal growth factor showed no significant difference. After 60 min the remaining radioactivity in the aged liver was half that in young liver. The total radioactivity secreted into bile was reduced from 16.5% +/- 5.7% to 10.0% +/- 2.3% of the injected dose in young and old animals, respectively. Per gram of liver and per 10(6) cpm injected, the old animals secreted only 30.9% of the amount secreted by young animals. The secretion of immunologically intact epidermal growth factor into bile was reduced from 1.6% +/- 0.8% to 0.78% +/- 0.2% of the injected dose in the young and old animals, respectively. Perfusion experiments showed that in aged liver the amount of total radioactivity secreted into the perfusate was increased to 133.6% that of the young liver. Per gram of liver and per 10(6) cpm injected, however, we found a reduction to 80.7%. Exocytosis at the canalicular and basolateral membrane appeared to be differently affected by aging. PMID- 8244270 TI - Regulation of tissue inhibitor of metalloproteinases-1 gene expression by cytokines and dexamethasone in rat hepatocyte primary cultures. AB - The steady-state levels of extracellular matrix proteins are regulated by the rates of their synthesis and degradation. Metalloproteinases and their specific inhibitors, tissue inhibitor of metalloproteinases-1 and -2 are believed to play a crucial role in extracellular matrix protein degradation. Here we show that the tissue inhibitor of metalloproteinases-1 is expressed in rat hepatocytes in primary culture and regulated by inflammatory cytokines. Rat hepatocytes constitutively express mRNA of tissue inhibitors of metalloproteinases-1 at a low level. Incubation with conditioned medium from lipopolysaccharide-stimulated human monocytes led to a dramatic induction of mRNA of tissue inhibitors of metalloproteinases-1. The inflammatory cytokines interleukin-1 beta, interleukin 6, interleukin-11, leukemia inhibitory factor and ciliary neurotrophic factor were also capable of stimulating expression of mRNA of tissue inhibitors of metalloproteinases-1. Among these cytokines interleukin-6 was the most potent stimulator. The combination of interleukin-1 beta, interleukin-6 and interleukin 11 synergistically up-regulated mRNA of tissue inhibitors of metalloproteinases 1. The synthetic glucocorticoid dexamethasone dose dependently inhibited constitutive and interleukin-6-induced expression of tissue inhibitors of metalloproteinases-1. A possible involvement of tissue inhibitor of metalloproteinases-1 in the pathogenesis of liver fibrosis and cirrhosis is discussed. PMID- 8244271 TI - Stimulation of liver growth by exogenous human hepatocyte growth factor in normal and partially hepatectomized rats. AB - Human hepatocyte growth factor stimulates DNA synthesis by cultured rat hepatocytes. When human hepatocyte growth factor prepared from the culture medium of human embryonic lung fibroblasts was intravenously injected into normal rats and rats after 70% hepatectomy, it was detected in hepatocytes but not in nonparenchymal cells isolated 30 min after injection. Similar injections of human hepatocyte growth factor at 2-hr intervals for 10 hr significantly increased hepatic DNA content in normal rats at 48 hr, with increased hepatic content of putrescine, the essential polyamine for hepatic DNA synthesis after 70% hepatectomy, and activities of catalytic enzymes of putrescine synthesis at 6 hr almost to the levels in rats after 70% hepatectomy. Those levels in rats after 70% hepatectomy were further enhanced by similar injections of human hepatocyte growth factor starting immediately after surgery. Increased hepatic DNA content in normal rats and rats after 70% hepatectomy was also seen with recombinant human hepatocyte growth factor to a greater extent compared with that seen with human hepatocyte growth factor. In normal rats given recombinant human hepatocyte growth factor, 5-bromo-2'-deoxyuridine-labeled and mitotic hepatocytes were significantly increased in number at 26 hr but not at 48 hr. We conclude that exogenous human hepatocyte growth factor acts as a trigger and a promoter of liver growth to increase hepatic putrescine production in rats. Recombinant human hepatocyte growth factor is more potent than human hepatocyte growth factor in this action. PMID- 8244272 TI - Degradation and intracellular accumulation of a residualizing hyaluronan derivative by liver endothelial cells. AB - The release and intracellular accumulation of 125I-hyaluronan degradation products was studied in cultured liver endothelial cells with hyaluronan oligosaccharides (relative molecular mass = approximately 44,000) uniquely modified and radiolabeled at the terminal reducing sugar. Two methods were combined to measure 125I-hyaluronan degradation by liver endothelial cells. (a) Cetylpyridinium chloride precipitation of hyaluronan oligosaccharides was used as a rapid, convenient assay to monitor the appearance of hyaluronan degradation products. Hyaluronan oligosaccharides less than 54 to 60 monosaccharides in length were not precipitated with cetylpyridinium chloride and thus were assessed as degraded. (b) Gel filtration chromatography was used to estimate the size range of oligosaccharides produced by liver endothelial cells. After internalization of 125I-hyaluronan, liver endothelial cells released radioactive degradation products into the culture media after a lag period of 2.5 to 3.0 hr. The intracellular accumulation of degraded 125I-hyaluronan was linear for at least 2 hr even though no degradation products were released. The long lag before release of degraded 125I-hyaluronan is likely caused by the modified chemical structure at the reducing end of the hyaluronan derivative; the derivative acts like a residualizing label. After this lag the release of degraded 125I hyaluronan proceeded linearly for up to 12 hr. The extracellular 125I-hyaluronan degradation products eluted with a distribution coefficient of 1.3 on a gel filtration column. The major intracellular 125I-labeled degradation product showed the same retardation (distribution coefficient = 1.3). This retention may be caused by the hydrophobic aromatic and alkyl modifications to the former reducing sugar, also characteristics of a residualizing label. In addition, at least two larger minor intermediates were observed intracellularly. The rate of intracellular 125I-hyaluronan degradation was dependent on hyaluronan concentration and reached a maximal rate (159 molecules/cell/sec) at 2 x 10(-7) mol/L. This was about half the maximal rate of endocytosis (285 molecules/cell/sec) at a hyaluronan concentration of 1.3 x 10(-7) mol/L. The apparent ligand concentration that gives half-maximal responses for endocytosis and intracellular degradation was 0.6 x 10(-7) and 1.0 x 10(-7) mol/L, respectively. PMID- 8244273 TI - Production of chemotactic factor, interleukin-8, from hepatocytes exposed to ethanol. AB - In a previous paper, we demonstrated proteinous chemotactic factors released from hepatocytes exposed to ethanol. In this study, we further characterized the chemotactic factors released from ethanol-treated hepatocytes. After fractionating the conditioned medium with gel chromatography, we demonstrated chemotactic activity at molecular weights of around 20 and 40 kD. When the conditioned medium was mixed with antiserum against interleukin-8, chemotactic activity was almost completely abolished. In addition, antiserum against interleukin-8 completely reduced chemotactic activity of the 20-kD chemotactic factor but did not influence the chemotactic activity of the 40-kD chemotactic factor. After gel electrophoresis of the conditioned medium, the protein reacted with antibody against interleukin-8 was demonstrated at a molecular weight of 20 kD but was not found at a molecular weight of 40 kD. These results suggest that a major part of the proteinous chemotactic factors released from hepatocytes exposed to ethanol could be a dimer form of interleukin-8, one of the proinflammatory cytokines, possibly contributing to the pathogenesis of alcoholic hepatitis. PMID- 8244274 TI - Role of ethanol-inducible cytochrome P-450 2E1 in the development of hepatocellular carcinoma by the chemical carcinogen, N-nitrosodimethylamine. AB - Cytochrome P-450 2E1 is a specific isozyme of cytochrome P-450 induced by ethanol. P-450 2E1 may also be the only enzyme that metabolizes N nitrosodimethylamine at a very low concentration. Because N-nitrosodimethylamine is a procarcinogen, the possibility that induction of P-450 2E1 by alcohol abuse may accelerate the carcinogenic action of a very small dose of N nitrosodimethylamine should be considered. To investigate this possibility, effects of ethanol on the hepatic carcinogenic action of a very small dose of N nitrosodimethylamine were analyzed in rats. Sixty Wistar male rats were divided into two groups (ethanol and control) according to the liquid diets they were fed. After preliminary feeding, N-nitrosodimethylamine in two concentrations (1.5 ng/ml and 3.0 micrograms/ml) in drinking water was given with the diets for 40 or 60 wk. To avoid metabolic competition between ethanol and n-nitrosodimethylamine, we gave N-nitrosodimethylamine and the diets alternately. The average amount of N nitrosodimethylamine consumed per day was 11 to 37 ng/kg body weight in the 1.5 ng group and was 27 to 37 micrograms/kg body weight in the 3.0-microgram group. Hepatic P-450 2E1 content was six times higher in the ethanol-treated groups than in the control groups throughout the experiment. Visible nodules and enzyme altered foci were not found in rats in any group at the end of the fortieth week. These preneoplastic changes were found at the end of the sixtieth week only in rats in the groups treated with both ethanol and N-nitrosodimethylamine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244275 TI - Differential impact of Carolina rinse and University of Wisconsin solutions on microcirculation, leukocyte adhesion, Kupffer cell activity and biliary excretion after liver transplantation. AB - This quantitative in vivo fluorescence microscopy study investigated the relative impact of an optimized rinse solution (warm Carolina rinse) and that of an established storage solution (University of Wisconsin solution) on various pathomechanisms of hepatic reperfusion injury after cold storage. Syngeneic orthotopic, arterialized liver transplantation was performed in male Lewis rats after 24 hr of cold ischemia (n = 24). The four experimental groups differed according to the type of preservation/rinse solution used: University of Wisconsin solution/albumin rinse (group 1), autologous blood (just external cooling)/albumin rinse (group 2), blood/Carolina rinse (group 3) and University of Wisconsin solution/Carolina rinse (group 4). Hepatic microvascular perfusion, leukocyte accumulation and phagocytic activity of Kupffer cells were assessed by means of intravital fluorescence microscopy 30 to 90 min after reperfusion. Disturbances of microvascular perfusion were most pronounced in group 2, markedly reduced by University of Wisconsin solution (group 1) and Carolina rinse (group 3) and minimized by combined use of University of Wisconsin solution and Carolina rinse in group 4. Intrahepatic leukocyte-endothelium interaction in sinusoids and postsinusoidal venules was found to depend on the application of Carolina rinse before reperfusion rather than the use of University of Wisconsin solution during cold storage. Activation of phagocytosis by Kupffer cells was most pronounced in group 1, intermediate in groups 2 and 3 and not noticeable in group 4. Hepatocellular excretory function as assessed on the basis of total bile flow and excretion of bile acids during the first 90 min after reperfusion was found to be improved by application of Carolina rinse, both after storage in blood or in University of Wisconsin solution.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244276 TI - Increased dietary fat content accelerates cholesterol gallstone formation in the cholesterol-fed prairie dog. AB - Epidemiological studies have provided conflicting information about the relationship between fat consumption and gallstone formation. We studied cholesterol gallstone formation in prairie dogs after 1 wk of the following diets: (group A) a control diet with no added cholesterol and 5% of calories from corn oil, (group B) 1.2% cholesterol with 5% of calories from corn oil or (group C) 1.2% cholesterol with 40% of calories from corn oil. In controls serum cholesterol was 58.9 +/- 4.5 mg/dl, gallbladder bile was unsaturated with cholesterol (cholesterol saturation index = 0.7 +/- 0.1; cholesterol = 3.8 mmol/L) and none of 12 animals formed cholesterol crystals or stones. The low-fat diet supplemented with cholesterol (group B) increased serum and biliary cholesterol concentrations to 292 +/- 76 mg/dl and 7.5 +/- 1.1 mmol/L, respectively (p < 0.05), but cholesterol saturation index was only modestly increased (1.1 +/- 0.1) and in only one of eight animals did cholesterol monohydrate crystals develop. Group C, animals, which received cholesterol plus high levels of corn oil, had higher serum cholesterol levels (457 +/- 66 mg/dl), higher biliary cholesterol concentrations (16.6 +/- 1.3 mmol/L), higher cholesterol saturation indexes (1.7 +/- 0.1) and increased incidence of cholesterol gallstones (5 of 11). The two cholesterol-supplemented diets increased biliary phospholipid concentrations, decreased the ratio of cholic/chenodeoxycholic acid and increased the proportion of biliary lecithins containing linoleic acid, but these abnormalities were greatest in group C, which was given large amounts of corn oil. These findings suggest that cholesterol gallstone formation in the prairie dog is accelerated by increased dietary omega 6 polyunsaturated triglycerides. PMID- 8244277 TI - Plasma elimination of indocyanine green in the intact pig after bolus injection and during constant infusion: comparison of spectrophotometry and high-pressure liquid chromatography for concentration analysis. AB - Indocyanine green is used to estimate liver blood flow rate and hepatic intrinsic clearance. However, its use as a test substance for studies of liver function has been limited by two puzzling kinetic observations: a biexponential plasma decay after bolus injection with an extremely slow late phase and an apparently steadily decreasing clearance value during constant infusion. These observations have been made with spectrophotometric concentration analysis. In anesthetized 30 to 40-kg pigs, we examined plasma concentration curves of indocyanine green after intravenous bolus injection and during long-term infusion. We compared spectrophotometry with high-pressure liquid chromatography for measurement of plasma indocyanine green concentration. In freshly prepared commercially available indocyanine green, high-pressure liquid chromatography could separately measure two fractions, the genuine indocyanine green (97% to 99% of total) and an in vitro degradation product (1% to 3%). Because their spectra were nearly identical, these fractions could not be distinguished by spectrophotometry. After intravenous administration both fractions were identified in the plasma by high pressure liquid chromatography. In the first series (n = 6) 25 mg of indocyanine green was injected intravenously for 5 min. When analyzed by high-pressure liquid chromatography, the genuine indocyanine green plasma concentration decay was biexponential with rate constants 0.196 +/- 0.021 (mean +/- S.E.M., n = 6) and 0.0372 +/- 0.0064 min-1. The degradation product of indocyanine green decayed almost monoexponentially, with a rate constant of 0.0093 +/- 0.0002 min-1. With spectrophotometry a biexponential decay was observed with rate constants 0.130 +/ 0.012 and 0.0095 +/- 0.0001 min-1. The biexponential decay of indocyanine green after spectrophotometry was the result of codetermination of the two fractions: genuine indocyanine green was responsible for initial phase, and the degradation product of indocyanine green was responsible for the late phase. In the second series (n = 9), indocyanine green was administered as a constant intravenous infusion. From 90 to 240 min the intrinsic hepatic clearance of genuine indocyanine green did not change detectably with time. In contrast, the degradation product of indocyanine green never reached steady-state concentrations. Because of code-termination of these two indocyanine green fractions, the apparent intrinsic hepatic clearance of indocyanine green estimated from spectrophotometry was steadily decreasing by 8.9% +/- 1% per hour of its initial value. At the same time estimation of liver plasma flow rate based on Fick's principle was not affected by the choice of analytical methodology. These observations indicate that high-pressure liquid chromatography is superior to spectrophotometry for kinetic analysis of indocyanine green elimination. PMID- 8244278 TI - Increase in prostanoid formation in rat liver macrophages (Kupffer cells) by human anaphylatoxin C3a. AB - Human anaphylatoxin C3a increases glycogenolysis in perfused rat liver. This action is inhibited by prostanoid synthesis inhibitors and prostanoid antagonists. Because prostanoids but not anaphylatoxin C3a can increase glycogenolysis in hepatocytes, it has been proposed that prostanoid formation in nonparenchymal cells represents an important step in the C3a-dependent increase in hepatic glycogenolysis. This study shows that (a) human anaphylatoxin C3a (0.1 to 10 micrograms/ml) dose-dependently increased prostaglandin D2, thromboxane B2 and prostaglandin F2 alpha formation in rat liver macrophages (Kupffer cells); (b) the C3a-mediated increase in prostanoid formation was maximal after 2 min and showed tachyphylaxis; and (c) the C3a-elicited prostanoid formation could be inhibited specifically by preincubation of C3a with carboxypeptidase B to remove the essential C-terminal arginine or by preincubation of C3a with Fab fragments of a neutralizing monoclonal antibody. These data support the hypothesis that the C3a-dependent activation of hepatic glycogenolysis is mediated by way of a C3a induced prostanoid production in Kupffer cells. PMID- 8244279 TI - Cholesterol carriers in human bile: are "lamellae" involved? AB - Cholesterol, a highly insoluble molecule, is transported in bile by specialized lipid aggregates. On the basis of extensive correlations between laboratory prepared model biles and surgically harvested native biles, it has become generally accepted that biliary cholesterol is solubilized by simple and mixed micelles, single bilayered (unilamellar) vesicles and, under certain conditions, multilamellar vesicles (liposomes or liquid crystals) all composed of bile salts, lecithin and cholesterol in different proportions. Current concepts suggest that in lithogenic biles multilamellar vesicles result from aggregation and fusion of unilamellar vesicles and are a principal source from which cholesterol precipitates to form gallstones. Recent reports now challenge the prevailing paradigm by proposing that the principal cholesterol-carrying particles in human biles are not micelles but are "lamellae" composed of stacked membrane-like bilayers of lipids. In this article, we provide a critical overview of the experiments that led to the established views of biliary cholesterol transport and to the newer lamellae hypothesis. The principal evidence for lamellae stems from negative-stain electron microscopy, an artifactprone technique when used to study lipid-rich fluids such as bile. We show that lamellar structures represent both the electron microscopic analog of multilamellar vesicles in supersaturated biles that presage the nucleation of cholesterol crystals and an electron microscopic artifact of fossilized mixed micelles that are in fact very tiny (2 to 4 nm in radius) by state-of-the-art noninvasive techniques. We argue further that the lamellae nomenclature improperly equates two fundamentally distinct physical-chemical mechanisms for cholesterol solubilization and dispersion in bile on the basis of identically appearing electron microscopic images. PMID- 8244280 TI - Stress proteins: is their application in clinical medicine on the horizon? PMID- 8244281 TI - Hepatocyte apoptosis: is transforming growth factor-B1 the kiss of death? PMID- 8244282 TI - DNA/protein complexes delivered in conjunction with adenovirus generate high efficiency in vitro transfection: a powerful transfection reagent with potential broad applications. PMID- 8244283 TI - Of windows, fenestrae and defenestrations. PMID- 8244284 TI - Normal increase in glucose output by portal noradrenaline and glucagon but not nerve stimulation with enhanced portal resistance, not only in micronodular but also in intermediate thioacetamide cirrhotic livers. PMID- 8244285 TI - [The effect of acupuncture on plasticity of spinal Clarke's nucleus synaptic terminals]. AB - Ten unilateral spared root cats with sectioned L1-S2 dorsal root ganglia, except L6, were used to explore the effect of acupuncture on plasticity of spinal Clarke's nucleus synaptic terminals. Five of the cats were needled on the operated hind limb for two courses (ten days as a course), and the other five were kept for control. Two pairs of acupoints located in the innervated area of L6 spinal nerve were used. The number of synaptic terminals per photo in neuropil of L3 Clarke's nucleus was compared directly, because there was no significant change in the area of Clarke's nucleus and its neuropil on the operated side of both groups. No significant difference was found in the number of three types of synaptic terminals between the unoperated sides of two groups, suggesting that acupuncture may not influence the contralateral side. In the control group, the number of giant axonal terminals from dorsal root of the operated side decreased to 75% of the unoperated side; however, in the experimental group, it recovered to contralateral or normal level of the operated side and was significantly higher than that of the control group's operated side, denoting that acupuncture may promote the plasticity such as collateral sprouting and synaptic reinnervation of coarse myelinated fibers from intact dorsal root, L6 mainly. The number of other two types of terminals from spinal interneurons showed no significant difference between the operated sides of two groups, indicating that acupuncture may not enhance the plasticity of nondorsal root nerve fibers. PMID- 8244286 TI - [Antigen analysis of McAb E4B7D5 directed against outer envelope of Leptospira interrogans serovar lai by SDS-PAGE and immunoblot]. AB - McAb E4B7D5 was prepared by hybridoma technology in BALB/c mice immunized to outer envelope of Leptospira interrogans serovar lai. This McAb agglutinated specifically with all the 13 serovars of Icterohaemorrhagiae serogroup in MAT test at high titres and protected the guinea pigs against the attack of virulent strain (017) of serovar lai. SDS-PAGE and immunoblot were used to analyse the reaction of the outer envelopes of the five strains of Leptospira (Leptospira interrogans Icterohaemorrhagiae serogroup, serovar lai, 017 and 601 stains; L. interrogans Hebdomadis serogroup, hebdomadis serovar strain 156; L. interrogans Australis serogroup australis serovar strain 620; biflexa leptospira patoc serovar strain Patoc I; with McAb E4B7D5. Results indicated that this McAb E4B7D5 recognized specifically 34.5 kd and 39.5 kd outer envelope proteins of serovar lai, strain 017 and 601, so it might be an immunoprotective McAb directed against Leptospira interrogans serovar lai. It was suggested that 34.5 kd and 39.5 kd outer envelope proteins of serovar lai recognized specifically by McAb E4B7D5 might be immunoprotective antigens. PMID- 8244287 TI - [Rat bladder ubiquitin-like molecule: isolation, purification and N-terminal sequencing]. AB - A Ubiquitin-like peptide was accidentally isolated from rat bladder by using 5% acetic acid wash while we were isolating antibacterial peptides. The purified molecule was obtained by reverse phase high performance liquid chromatography. Gas phase microsequence analysis indicated the N-terminal sequences of the molecule as follows: MET-GLN-ILE-PHE-VAL-LYS-THR-LEU-THR-GLY-LYS-THR-ILE-THR-LEU- GLU-VAL-GLU-PRO-SER-ASP-THR-ILE-GLU-ASN, which is homologous to human ubiquitin. Ubiquitin plays a role in the differentiation of pre-B lymphocytes, Thus, it is suggested from the findings of this molecule and the endogenous antibacterial polypeptides in mucosa or mucosal epithelium that mucosal epithelium also might be one of immune cells or immunity-associated cells, which may secrete effector molecules directly to kill adherent microbes and produce regulating factors in mucosal immune response. PMID- 8244288 TI - [Effects of high-carbohydrate diet on VLDL-, HDL-, and apolipoprotein CII receptor activities on liver parenchymal and non-parenchymal cells in rats]. AB - We studied the functions of hepatic VLDL-, HDL-, and apolipoprotein (apo) CIII receptors in the endogenous hypertriglyceridemic (HTG) rats induced by feeding a high-carbohydrate diet (HCD) (80% of total calories). Compared with the control rats (60% of total calories in CHO), the fasting plasma triglyceride (TG) levels of the rats fed on the CHO diet for 5 days increased by 113.4% (P < 0.025, n = 6), but HDL-C and total cholesterol concentrations decreased by 28.4% and 14.4% (P < 0.05, P < 0.05). The binding (Bmax) and uptake of 125I-labeled VLDL by liver non-parenchymal cells (NPC) from HTG rats decreased by 65.0% and 19.1% (P < 0.001, P > 0.05) respectively. The binding of 125I-labeled apoE-deficient HDL3 to the NPC also decreased significantly (-54.5%, P < 0.05). But the affinities (Ka) of the two receptors were prone to increasing. However, the activities of VLDL- and HDL-receptors on liver parenchymal cells didn't significantly change. Meanwhile, the specific binding of 125I-labeled apo CIII to NPC from HTG rats increased by 104.5% (P < 0.025). The results of this study suggest that hepatic VLDL- and HDL-receptors could be down- and up-regulated respectively in response to plasma lipoprotein concentrations in the high CHO dietary rats, and apo CIII receptor might play an important role in regulating VLDL- and HDL-receptor activities. PMID- 8244289 TI - [A study on the transcriptional expression of oncogenes in human primary brain tumors]. AB - The c-myc, L-myc, N-myc, Ha-ras, c-fos, and v-erbB oncogenes were used as probes to detect the total RNA of 5 cases of normal human brain tissues and 29 cases of brain tumor tissues by RNA dot hybridization analysis. The results showed that the increasing expressions of cerbB and c-fos were the highest, being 88.2% and 82.4%, respectively, the next was c-myc with an increased expression of 46.1%; the increasing expressions of L-myc (29.3%) and N-myc (31.6%) were observed in a few samples only. There was no significant difference of Ha-ras gene expression between normal brain and brain tumor. In addition, we observed the increasing expression of more than one oncogene in one sample. These results suggest that c erbB and c-fos oncogenes may play an important role in the pathogenesis of human primary brain tumors, and that multiple genetic changes are implicated in genesis of human brain tumors. PMID- 8244290 TI - [Electrophoretic analysis of isoenzyme and protein patterns of Romanomermis yunanensis and R. culicivorax (Nematoda:Mermithidae)]. AB - Isoenzyme electrophoretic patterns of lactate dehydrogenase (LDH), malate dehydrogenase (MDH), esterase (EST) and peroxidase (PO) of post-parasitic juveniles and adults of Romanomermis yunanensis and R. culicivorax were analysed by disc electrophoresis, respectively. The protein patterns of the two species of Romanomermis were compared by vertical slab SDS polyacrylamide gel electrophoresis. The results showed marked differences in isoenzyme patterns (number of bands and/or Rf) of LDH and MDH between the two species of Romanomermis, but the similar positions of isoenzyme bands denoted a certain genetic relationship between them. No band was found in the isoenzyme patterns of PO and EST. Distinct differences in protein pattern were observed between the two species of Romanomermis, but some common bands in pattern reflected the phylogenetic relationships of these species. The differences of isoenzyme and protein patterns observed in this study have provided the reason for identifying and differentiating the two species of Romanomermis at the molecular level. PMID- 8244291 TI - [Respiratory effects of tutin microinjection into NPBM of rabbits]. AB - On sodium pentobarbital anesthetized, cervical vagotomized adult rabbits, the microinjection of 1 microliter (10 micrograms) tutin into NPBM causes reversible changes in electrical activity of the diaphragm in different degrees. The most remarkable change is the reversible apnea. Our results indicate that NPBM plays an important role in the generation or modulation of respiratory rhythm and that tutin stimulates the pontine respiratory related neurons, thus disturbs the activity of neuronal circuit involved in respiration rhythm generation or modulation. PMID- 8244292 TI - [Synchronous investigation of the effects of vesnarinone on action potential and muscle tension of isolated rabbit's ventricular papillary muscle]. AB - This study adopted intracellular microelectrode technique to observe synchronously the effects of vesnarinone on intracellular potential and muscle tension of isolated rabbit's ventricular papillary muscle and analysed their variation. The results showed that vesnarinone increased the Tmax (P < 0.01) and Tdv/dt (P < 0.01) of isolated rabbit's ventricular papillary muscle and prolonged the action potential time (APT) without obvious effects on RP, APA and Vmax of papillary muscle. The relationship between them (r = 0.994, P < 0.05), suggesting that their changes be of the same mechanism and probably relate to the increase of inward calcium current. PMID- 8244293 TI - [Analysis of estrogen receptors in normal bone and bone tumor tissues]. AB - Thirteen human normal bone and fifteen bone tumor tissues were assayed for estrogen receptor (ER) by Dextran-coated Charcoal method (Scatchard plot). The results showed that the concentrations of ER in normal bone tissues (15.12 +/- 14.68 fmol/mg pro) were higher than that of bone malignant tumors (8.04 +/- 6.71 fmol/mg pro) P < 0.05). The binding dissociation constants (Kd) of ER in bone tumors (18.46 +/- 27.10 x 10(-11) mol/L) was lower than those in normal ones (39.91 +/- 20.13 x 10(-11) mol/L) (P < 0.025). The percentages for positive score of ER in normal and tumor ones (23.1%, 26.7%) were not significantly different (P > 0.05). Our study indicated that the variations of concentration and affinity of ER in bone related to the tumor incidence. That cytosol ER content was decreased in malignant bone tumors whereas that of affinity increase suggests an impairment or change of an intact receptor mechanism in this tissues. PMID- 8244294 TI - [Anti-implantation action of nylestriol and its effect on uterine cytosol estrogen and progesterone receptor in rabbits]. AB - The anti-implantation action of nylestriol in rabbit was studied. The changes of uterine cytosol estrogen and progesterone receptor (ER and PR) were analysed with the dextran-coated charcoal method. Plasma progesterone level of early pregnant rabbits was measured by radioimmunoassay. The results showed that oral nylestriol could obviously prevent the early pregnancy of rabbit by anti-implantation of blastocyst. The content of ER decreased in the nylestriol group (P < 0.01). The content of PR in nylestriol group was lower than that of the control group (P < 0.05). No significant difference of plasma progesterone level was noted between the control and treatment groups. These results suggest that there is close relationship between the anti-implantation effect of nylestriol and the decreased content of cytosol ER and PR during the pre-implantation period. PMID- 8244295 TI - [Monoclonal Anti-Id as substitute for HBsAg to detect Anti-HBs]. AB - According to Jerne's idiotype-anti-idiotype network theory, a new method to detect Anti-HBs was studied. This method utilized monoclonal Anti-Id as a substitute for HBsAg of the routine method adopting the enzyme-linked immunosorbent assay (ELISA) format. The serum specimen was incubated with solid phase (polyvinyl chloride) that had been coated with monoclonal Anti-Id. If Anti HBs was present in the serum, it would be combined with the Anti-Id. HRP-labeled goat Anti-human IgG was added, which reacted with the immobilized Anti-HBs. Under optimal conditions, 0.5 microgram/ml Anti-HBs could be detected. This method is safe, simple, and specific for Anti-HBs with no cross-reactivity with Anti-HBs, Anti-HBe and rheumatic factor positive serum, and the reagents can be obtained in unlimited amounts and are homogeneous. These features are particularly attractive when the antigen is difficult to obtain. PMID- 8244296 TI - [Computer analysis of evoked field potentials in hippocampal slice]. AB - A microcomputer system for the processing of evoked field potentials of brain slices was developed on the IBM PC-AT microcomputer. In the system, a 385-point finite impulse response (FIR) filter was designed and fast Fourier convolution algorithm was adopted to attain both high quality of filtering and high efficiency of calculation. Also a method for wave peak recognition by analysing waveform slope was established and an interface circuit was designed for the automatic control of the FM tape recorder by the computer. As a result, automated parameter measurement of epileptiform activity recorded from hippocampal slices was successfully realized with high accuracy. PMID- 8244297 TI - [Effects of interfacial state and bonding on the biostrength of plat castable ceramics crown]. AB - Plat castable ceramics (PCC) restorations depend primarily on their biostrength. In our experiment the human oral physiological environment was simulated and we studied the effects of restoration-tooth interfacial state and bonding on the biostrength of PCC crowns. The bond strength affects the biostrength of PCC crowns obviously. The interfacial treatment affects not only the bond strength between PCC crown and tooth structure but also the biostrength of the crown cemented. PMID- 8244298 TI - [Morphometric evaluation on the pathologic changes in ovariectomized endometria influenced by estrogen use]. AB - Thirty SD female rats were divided into the following groups: (1) sham operation; (2) bilateral ovariectomized (OVX); (3) OVX and estrogen-treated (E). Computerized Image Analyser was used to evaluate the pathologic changes of endometria in these animals. The results showed the endometria of OVX group was in the state of atrophia, while the endometria of E group was in the state of early proliferative stage. The heights of endometria, endometrial epithelia cells and gland epithelia cells, the volume fraction and numerical density of endometrial glands, and the cavity diameter of the glands could effectively express the proliferative changes of the endometria. It is suggested that this method be used to study the pathology of the different endometric diseases. PMID- 8244299 TI - [Pathologic observation of 298 rat mesotheliomas induced by asbestos]. AB - Two hundred and ninety eight rat mesotheliomas induced by chrysotile or crocidolite were studies. The results demonstrated that tumor masses were scattered extensively. The appearance of tumor was polymorphous. According to the microscopic features, the 298 cases of mesothelioma could be histologically classified into three types, i.e. epithelial type, fibrous type and mixed cell type. The malignant mesothelioma is characteristic of its polymorphism and the clear cancernation of mesothelial cells. PMID- 8244300 TI - [Isolation and purification of a NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase from human placenta]. AB - A NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase has been isolated and purified from human placenta. About 9300-fold enrichment was achieved with a yield of 9.3%. The specific activity of the purified enzyme was 6470 nmol/min.mg. 15-OH-PGDH showed a single band on PAGE and two bands on SDS-PAGE. The molecular weights were approximately 26 and 52 kd, respectively. The two bands were recognized by both polycloning and monocloning antibodies of the enzyme. It is suggested that the 15-OH-PGDH be composed of two identical subunits. PMID- 8244301 TI - [Study on determination of polyactin A by ELISA]. AB - Sandwich ELISA and two-step competition ELISA were utilized to determine polyactin A. After optimization of the experimental conditions, microtiter plates were coated with the rabbit antibody solution and evaporated to dryness at 37 degrees C. The coated microtiter plates were incubated first with the analyte, then with the HRP-labelled antibody (sandwich procedures), or with the HRP labelled polyactin A (competition procedures), and absorbance was measured after coloration. The whole determination took less than 4 hours, excluding the coating procedure, which is faster than the routine complement fixation method. The coated microtiter plates can be well after over three months of storage at 4 degrees C. The sensitivity obtained was 0.64-7.4 ng/well for the sandwich method or 0.6-1.2 micrograms/well for the competition method. PMID- 8244302 TI - [The applicability of a new stable fluorescence reagent CGE (N) for TLC measurements and spectrofluorimetry]. AB - A new stable fluorescent reagent, CGE (N), was designed and synthesized in our laboratory. This paper presents the results of using it to detect a compound having aminogroup. Glycine as an example was derived by using CGE (N), the best condition of derivatisation reaction was pH8, T 50 +/- 1 degree C, and time 3-5 h. Spread agent was CHCl3: CH3OH:CH3COOC2H5: iceHAc (1:3:5:1). Rf of glycine-CGE (N) was 0.45, of which fluorescent spectrum Em was 356nm (at Ex = 277nm). The correlation coefficient between the fluorescent scanning area and quantity was over 0.99, the linear relation retained for 24 h, the detection limit 0.24 microgram, and the linear range 0.24-19.2 micrograms. The data revealed that stability, sensitivity and reproducibility were good. These indicate that CGE(N) is a fine fluorescent reagent for compounds having primary amino-group and this method is reliable. PMID- 8244303 TI - [Effects of air pollution on health of residents in vicinity of an electrometallurgical factory in Chengdu]. AB - Air pollution in an electrometallurgical plant in Chengdu was monitored in 1988. Results showed that atmospheric TSP and Ni concentrations in the vicinity of the plant were higher than those in control area; and TSP content higher than its health standard level. A population of 262 children was physically examined in the same pollution and control areas in May, 1988. Physical examinations were as follows:otorhinolaryngological examination, skin patch test of Ni and Co, immune function test (including PHA skin test, salivary LZM content), Ni levels in human hair and urine etc. We found that body burdens of Ni in population of pollution area increased because of the pollution of Ni compounds in the atmosphere. Hair Ni content in pollution area was significantly higher than that in control area, and the positive rate of Ni skin patch test was also markedly higher than that in control area. Case history indicated that there was a significant difference in Ni content as detected by otorhinolaryngological examinations between the pollution and control areas. Some measures were suggested to improve the air quality based on the study. PMID- 8244304 TI - [Study of teratogenicity and sensitive period of vanadium pentoxide in Wistar rats]. AB - Pregnant Wistar rats were administered with vanadium pentoxide (V2O5) in three different ways single intraperitoneal injection (ip) of V2O5 5mg/kg on days 9, 10, and 11 of gestation; ip 5mg/kg/day on days 9-12 of gestation; and ip 3mg/kg/day on days 6-15 of gestation. The results showed that the female rats had such toxic symptoms as decreased weight gain, increase fetal mortality, decreased fetal weight and crown-rump, retarded ossification of the bone, increased incidence of subcutaneous hemorrhage, rib wavy, dilation of lateral ventricles and renal pelvis in the two groups, i.e., on days 6-15, and 9-12 of gestation, Decreased weight gain, increased incidence of subcutaneous hemorrhage and visceral anomalies were found on day 11 of gestation group. Increased incidence of fetal death, retarded ossification of the bone on day 10 of gestation group; and increased incidence of subcutaneous hemorrhage and visceral anomalies on days 9, and 10 of gestation were observed. These data suggest that V2O5 can induce teratogenicity in Wistar rat with or without obvious maternal toxicity. There was no obvious teratogenic sensitive period of V2O5 on embryo, but as for single treatment, it was more sensitive on gestation day 10 than on gestational day 11 or 9; as for multi-treatment, it was more sensitive on days 9-12 than on days 6 15 of gestation for teratogenicity and fetal death. It is suggested that V2O5 may act mainly on the middle or late stage of organogenetic period. PMID- 8244305 TI - [Study on degradation of phenols in water]. AB - The degradation of phenols in water and the effects of microbiolism, pH and dissolved oxygen (DO) on the degradation were investigated. The results showed that the degradation of phenols in water was mainly that of biochemistry, which depends on the existence of microbiolism. The most suitable pH for the degradation was 6 to 9, and no effects of DO on it were found. The accustomization of microbiolism with phenols would accelerate the degradation. PMID- 8244306 TI - [Gas chromatographic analysis of nicotine in indoor air]. AB - An analytical method for the determination of nicotine in indoor air was developed. Air sample was collected in absorption solution of 1% NaHSO4. After alkalization and heptance extraction, the content of nicotine in the sample was determined by gas chromatography with FID. The average sampling efficiency was 93.6%. The recovery rate was 89.3%-105.5% (mean = 98.4%). The coefficient of variation was 1.7-4.1%. Under the sampling condition the detection limit was 0.01 microgram/m3. After discussing the stability and extraction efficiency of the sample, we found the air sample collected in the absorption solution was stable for at least 7 days in refrigerator. The field experimental results show that the method is simple, reliable and suitable for collecting and determining trace nicotine in indoor air. PMID- 8244307 TI - [Determination of serum pancreatic polypeptide and its clinical significance in patients with end state renal failure]. AB - Serum levels of pancreatic polypeptide (SPP) responding to 75g glucose (oral) challenge were observed in 27 patients with end state renal failure (ESRF) and 27 normal subjects as a control. The results showed that the levels of SPP were significantly higher in ESRF patients than in normal subjects. Dialysis therapy seemed to have no effect on the levels of SPP. The elevated levels of SPP could return to normality after kidney transplantation. There was no positive correlation between the levels of SPP, the blood sugar, and the serum insulin. It is suggested that kidneys are the important sites for the metabolism of PP. The elevated SPP concentration in patients with ESRF may result from their renal failure, and may contribute to some gastrointestinal symptoms which are frequently associated with ESRF. PMID- 8244308 TI - [Helicobacter pylori associated gastritis: a primary study of ammonia and urea in gastric juice and mucus in gastric mucosa]. AB - The mucosa of 46 patients with histological chronic gastritis were cultured for helicobacter pylori (HP) and the ammonia and urea in their gastric juice were also determined. The results showed the average ammonia concentration (1.22 +/- 0.23 mmol/L) in HP positive patients was higher than that of HP negative patients (0.72 +/- 0.25 mmol/L), (P < 0.05). But the average urea concentration (0.45 +/- 0.29 mmol/L) in HP positive patients was lower than that of HP negative patients (2.71 +/- 1.20 mmol/L), (P < 0.05). The higher the level of ammonia stands, the more severe the gastritis is. (rs = 0.556). On the other hand, the quantity of mucus was elevated in 18 patients among 28 patients with HP eradicated by drug treatment, which was significantly different from the patients with HP not eradicated. It is suggested that the epithelium of gastric mucosa and the mucus are the chief components of the gastric mucosa barrier; the presence of HP increases ammonia, and thus damages the gastric epithelium and reduces the quantity of mucus. Consequently, the protective gastric mucosa barrier is damaged, and then comes the liability to HP-associated gastritis. PMID- 8244309 TI - [The changes in free radical and prostaglandin metabolism in newborn infants with asphyxia]. AB - Blood plasma thromboxane-B2 (TXB2) and 6-keto-PGF1 alpha levels were determined by radioimmunoassay technique during the first six to sixteen hours of life in 16 newborn infants with severe asphyxia, 18 newborn infants with mild asphyxia and 27 normal term neonates. Plasma lipid peroxidation (LPO) content was measured by TBA-colour-contrast method in 15 infants with severe asphyxia, 17 infants with mild asphyxia and 24 healthy term newborn infants. The results showed that blood plasma LPO, TXB2 and 6-keto-PGF1 alpha levels in infants suffering from severe asphyxia were higher than those in infants with mild asphyxia and normal infants (P < 0.01), but no significant difference was noted between the mild asphyxia group and normal control group (P > 0.01). These suggest that production of free radicals is increased and prostaglandin metabolism is triggered in the infants with severe asphyxia, that cerebral ischemia and hypoxia caused by asphyxia contributes to the augmented production of prostaglandins and free radicals, and that accumulation of free radicals and prostaglandin enhances brain damage and the metabolism of arachidonic acid appears to be an important source of the free radicals in infants with intrauterine asphyxia. PMID- 8244310 TI - [Memory function of patients with cerebral arteriosclerosis evaluated by Rivermead behavioural memory test]. AB - Using Rivermead behavioural memory test, we examined 142 patients with cerebral arteriosclerosis. The results showed that the rate of anomalies of screening score in patients with cerebral arteriosclerosis were significantly higher than that of the control group (P < 0.01). The patients and control scores were significantly different (P < 0.05) in all items except the picture and face recognition. Comparing the screening score with brain CT scan, we found a relationship between brain damage and behavioural memory declination. Also, many persons of control group got low scores with behavioural memory test, suggesting that the Rivermead behavioural memory test be sensitive to memory declination. With its simplicity and sensitivity the Rivermead behavioural memory test may have some practical value in China. PMID- 8244311 TI - [Effects of "he xiang zhuang gongfu" on respiratory function in healthy adults]. AB - Eleven healthy adults who practised He Xiang Zhuang Gongfu (HGF) were followed for 6 months; 15 pulmonary function tests were determined before HGF practice, 2 months and 6 months after HGF practice. All 11 subjects took the HGF exercise every day for 30 minutes. Samplings were taken before subjects had ever practised HGF as control value, immediately and 30 minutes after HGF exercise. In the 2nd month and 6th month of HGF practice. The results indicated that VO2 decreased after 2 and 6 months HGF practice, and the effect even persisted to 30 minutes after HGF exercise, indicating that HGF may reduce metabolic rate and decrease the oxygen consumption of body. The possible mechanism was discussed. MVV had an up tendency after HGF practice, which suggested the possibility of strengthening respiratory muscle by HGF. HGF may be a good physical exercise and can induce a wakeful hypometabolic physiologic state. PMID- 8244312 TI - [The changes in the levels of angiotensins and angiotensin converting enzyme and effects of captopril in patients with hypoxic pulmonary hypertension]. AB - The levels of angiotensin I, II (ATI, II) and angiotensin converting enzyme (ACE) were measured in 28 patients with hypoxic pulmonary hypertension and 23 normal individuals. The effects of captopril were observed in 8 of 28 patients. The results showed that the average level of ATII in the patients was significantly higher than that in normal control (P < 0.05), but the average level of ACE in the patients was significantly lower than that in the controls (P < 0.05). In 8 patients, after the use of captopril, an ACE inhibitor, for two weeks, the levels of ATII, ACE and mean pulmonary arterial pressure (mPAP) were markedly decreased (P < 0.05). But the other 20 patients treated with antibiotics, antiasthmatics, diuretics and oxygen therapy for two weeks did not show any alterations in the levels of ATII, ACE and mPAP. PMID- 8244313 TI - When is a new diagnostic method established? PMID- 8244314 TI - Distinction of follicular hyperplasia from follicular lymphoma in B5-fixed tissues: comparison of MT2 and bcl-2 antibodies. AB - The distinction between follicular hyperplasia (FH) and follicular lymphoma (FL) is one of the classic problems in diagnostic hematopathology. Both MT2 and bcl-2 antibodies have been reported to be useful in making this distinction. To compare the utility of these two commercially obtained antibodies in paraffin-embedded sections, B5-fixed sections from 21 cases of follicular hyperplasia and from 46 follicular lymphomas were immunostained and the follicles evaluated. Neither MT2 nor the bcl-2 antibody showed definite follicular positivity in FH. In contrast, 66% of FLs were MT2 positive and 89% were bcl-2 positive (P < .02). MT2 staining was equivocal in 30% of FHs and 7% of FLs, whereas bcl-2 staining was equivocal in only one FL. In addition, MT2 staining was unsatisfactory in three cases. CD10 was negative (0% to 3%) in the three bcl-2-negative FLs tested and positive (> 10%) in 26 of 38 bcl-2-positive cases. These results demonstrate the utility of the commercially obtained bcl-2 antibody in distinguishing FL from FH in B5 fixed, paraffin-embedded tissue sections. They also illustrate the superiority of the bcl-2 antibody over MT2, which was less sensitive and more problematic to interpret. PMID- 8244315 TI - Sulfur-rich prostatic intraluminal crystalloids: a surgical pathologic and electron probe x-ray microanalytic study. AB - Prostatic intraluminal crystalloids are irregular eosinophilic, non-birefringent structures increasingly recognized as potential indicators of prostatic malignancy. In a study of 250 randomly selected surgical pathology cases of prostatic tissues none of 50 cases of benign glandular hyperplasia (0%), one of 50 cases of atypical adenomatous hyperplasia, 18 of 50 cases of well differentiated adenocarcinoma (36%), seven of 50 cases of moderately differentiated adenocarcinoma (14%), and none of 50 cases of poorly differentiated adenocarcinoma (0%) revealed intraluminal crystalloids. A histochemical and immunohistochemical staining panel indicated that the crystalloids were nonproteinaceous. Crystalloids were stained intensely with phosphotungstic acid hematoxylin and did not stain for prostatic-specific antigen or hemoglobin. Crystalloids were clearly differentiated from prostatic corpora amylacea on light microscopy, histochemistry, scanning, and transmission electron microscopy. Electron probe x-ray microanalysis of 10 cases of crystalloids revealed uniform high sulfur peaks and small sodium peaks. We conclude that intraluminal crystalloids are associated more frequently with low-grade prostatic adenocarcinoma, may occur in benign tissue bordering adenocarcinoma, are predominantly composed of inorganic sulfur, and their presence in benign and atypical prostate biopsies may be of pathologic significance and should warrant further clinical investigation and possibly repeat biopsy. PMID- 8244316 TI - Juxtaglomerular cell tumor of the kidney: report of two cases with a papillary pattern. AB - We report the clinicopathologic, immunohistochemical, and electron microscopic study of two cases of juxtaglomerular cell tumor of the kidney with a hitherto unreported dominant papillary pattern. Both tumors were associated with high blood pressure that did not respond to medical therapy, but that returned to normal after removal of the kidney. They were well delineated, tan, and had no necrosis. The cores of the papillary structures consisted of polygonal cells found to express renin by immunohistochemistry and to contain renin protogranules by electron microscopy. The papillary fronds were covered by one layer of cuboidal epithelial cells that did not stain for renin and had ultrastructural features reminiscent of the collecting duct epithelium. These tumors must be differentiated from malignant papillary tumors of the kidney, such as papillary clear cell carcinoma, transitional cell carcinoma, and collecting duct carcinoma. PMID- 8244317 TI - p53 accumulation in ovarian carcinomas and its prognostic implications. AB - Intranuclear p53 accumulation is a common finding in many different human tumors and is associated with a worse prognosis in breast, colon, and lung carcinomas. We immunostained a series of common epithelial ovarian cancers to assess (1) the prevalence of p53 accumulation and its clinicopathologic correlations, and (2) its prognostic implications. The study population comprised 98 patients (83 with follow-up data). A variable degree of p53 immunoreactivity, strictly confined to the neoplastic cells, was detected in 54 tumors (55%). Among these tumors there were 10 low expressors (1% to 10% immunoreactive tumor cells), 16 moderate expressors (10% to 50% immunoreactive cells), and 28 high expressors (> 50% immunoreactive cells). The prevalence of p53 immunoreactivity did not show any association with the histologic type of the tumors or with the disease stage at presentation. p53 Accumulation, however, was significantly more prevalent among poorly differentiated tumors (P = .0059, by analysis of variance). Life table analysis demonstrated that patients with tumors showing moderate and high p53 expression had worse disease-free and adjusted lengths of survival (P = .0011 and P = .0025, respectively, by Mantel-Cox). The patients with "early stage" disease (stages I and II) and p53 accumulation showed a trend toward shorter length of survival, but this did not reach statistical significance. However, patients with "advanced stage" disease (stages III and IV) and moderate or high p53 accumulation had a poorer prognosis (P = .0154 and P = .0171, for disease-free and adjusted length of survival, respectively). These results suggest that p53 accumulation occurs more frequently in tumors with aggressive behavior and that p53 immunoreactivity may have a prognostic role in certain subsets of patients with ovarian carcinoma. PMID- 8244318 TI - Bronchial epithelial changes associated with small cell carcinoma of the lung. AB - From 1976 to 1991, 151 cases of small cell carcinoma of the lung were diagnosed by fiberoptic bronchoscopic biopsy at our institution. One hundred twenty-eight of 151 cases provided suitable material for the examination of the morphologic changes in the bronchial surface epithelium. Thirty-seven percent of the cases showed normal bronchial epithelium, 47% showed benign squamous metaplasia, 9% showed atypical squamous metaplasia, and 5% showed squamous cell carcinoma in situ. Immunohistochemical examination for bombesin and epidermal growth factor was performed on selected biopsy specimens. The biopsy specimens chosen for immunohistochemistry included 20 specimens that showed normal bronchial epithelium, 20 specimens with benign squamous metaplasia, 12 specimens with atypical squamous metaplasia, and seven specimens with squamous cell carcinoma in situ. All the specimens showed positive staining with anti-bombesin. With anti epidermal growth factor 10% of biopsy specimens with normal epithelium showed positive staining. The positive reaction increased from 25% for biopsy specimens with benign squamous metaplasia to 58% for biopsy specimens with atypical squamous metaplasia and to 71% for biopsy specimens with carcinoma in situ. These findings suggest a connection between epidermal growth factor production by small cell carcinoma of the lung cells and changes in the bronchial surface epithelium. PMID- 8244319 TI - The detection of B-cell monoclonal populations by polymerase chain reaction: accuracy of approach and application in gastric endoscopic biopsy specimens. AB - The recently developed strategy for the detection of monoclonal B-cell populations, based on the selective amplification of predominant immunoglobulin heavy chain gene (IgH) rearrangement, is seen to be a suitable alternative to Southern blot analysis. The new technique uses a pair of consensus primers for variable (VH) and joining (JH) regions. We first tested the accuracy of this new approach on a broad series of 67 samples that had been well characterized by both Southern blot and immunohistochemical techniques before being subjected to blind testing. Our results show that this technique gave 100% specificity (absence of false-positive results) and approximately 70% sensitivity (30% false-negative results). The only exception was the presence of an IgH polymerase chain reaction ambiguous result in a case of Sezary's syndrome. The polymerase chain reaction technique was then applied to a panel of 27 frozen gastric endoscopy biopsy specimens following previous clinical suspicion of lymphoma. Monoclonality was detected in nine of 13 samples previously diagnosed as lymphomas and in one of two carcinomas. Further examination of the gastrectomy specimen in the latter case disclosed a B-cell lymphoma associated with the carcinoma. In spite of its limited sensitivity, the high specificity attained by this technique in the detection of monoclonality makes it a useful adjunct to routine morphologic criteria, as it is sometimes capable of detecting true positive cases that conventional morphology studies show to be negative. PMID- 8244320 TI - Histopathologic features in the central nervous system of 400 acquired immunodeficiency syndrome cases: implications of rates of occurrence. AB - Histopathologic lesions in the central nervous system (CNS) of 400 autopsy cases of the acquired immunodeficiency syndrome (AIDS) collected from 1982 to 1990 were studied. Lesions most closely associated with human immunodeficiency virus (HIV) infection in the CNS (perivascular macrophages, nodular encephalomyelitis, diffuse leukoencephalopathy, necrotizing encephalitis, and long-tract degeneration) were found in 20% of the cases. The group of vascular and inflammatory lesions and of opportunistic infections was seen in 25% of cases. These two lesion groups were found together in 32% of cases, and none of these lesions was present in 23% of cases (most of the latter having no significant CNS lesions). Length of survival increased in the last group of 100 cases compared with the first 300 cases. The homosexual and bisexual risk groups showed continuously increasing lengths of survival for each category of HIV-associated CNS lesions throughout the study, while the lengths of survival in the other risk groups varied. Patients in the last group of 100 autopsy cases with any HIV associated lesion survived longer than patients without these lesions. The AIDS patients with no CNS lesions had the shortest mean length of survival. The results suggest that although survival is prolonged as specific therapy is given, there is an increase in CNS lesions in AIDS patients with longer survival. This may indicate that CNS lesions in AIDS are generally dependent on systemic disease progression over many months as immune function decreases. PMID- 8244321 TI - The pagetoid variant of urothelial carcinoma in situ. AB - The frequency, distribution, and clinical significance of pagetoid changes in urothelial carcinoma in situ (CIS) were studied in over 600 tissue slides from 102 patients followed for a mean period of over 4 years. Results indicate that pagetoid changes are unusual, randomly distributed, and never widespread. These changes occur in a histologic setting of CIS and apparently never appear as the initial lesion, even in cases in which CIS is the primary or only neoplasm. Patients manifesting pagetoid CIS had essentially the same progression and survival rates as patients without pagetoid changes. Our data suggest that this histologic pattern represents a peculiar reaction to injury confined to small areas of urothelium in susceptible hosts. PMID- 8244322 TI - Infantile histiocytoid cardiomyopathy--myocardial or conduction system hamartoma: what is the cell type involved? AB - Primary myocardial diseases in the pediatric age group encompass a variety of metabolic, infectious, congenital, and acquired disorders, one of which is "histiocytoid cardiomyopathy." We describe clinical and pathologic features in two infants. Autopsy findings in the first case were consistent with sudden cardiac death. The second infant has survived for 2 years on antiarrhythmic therapy with amiodarone. Microscopically, cells with vacuolated to granular cytoplasm were grouped in fascicles, imparting a pseudonodular appearance, but following a distribution reminiscent of conduction fibers. Ultrastructurally, the cells lack a T-tubule system, possess scattered lipid droplets and desmosomes rather than side-to-side junctions, and contain leptomeric fibrils that predominantly marginate to the cell periphery without sarcomeres. Immunostaining of paraffin-embedded tissue reveals perimembranous immunoreactivity for muscle specific actin, but not for the histiocytic markers CD68 (KP1) and lysozyme. Immunohistochemistry may be a practical alternative when tissue is not saved for electron microscopy. The clinical and pathologic features of this disorder in light of the current literature suggest that it may be hamartoma, possibly of conduction system origin. PMID- 8244323 TI - The value of polyclonal carcinoembryonic antigen immunostaining in the diagnosis of microvillous inclusion disease. AB - Microvillous inclusion disease is a specific disorder recognized as a cause of intractable diarrhea of infancy. We studied three cases by light microscopy, electron microscopy, and immunostaining for polyclonal carcinoembryonic antigen (CEA). Histologically, all cases had villous atrophy and abnormal accumulation of periodic acid-Schiff-positive material in surface enterocytes. Ultrastructurally, poorly developed brush-border and intracytoplasmic inclusions lined by intact microvilli were present in surface enterocytes. Crypt cells showed well-preserved surface microvilli. Carcinoembryonic antigen immunostaining showed prominent intracytoplasmic reactivity in surface enterocytes and linear brush-border reactivity in crypt cells. Normal and diseased small bowel biopsy specimens used as controls revealed linear brush-border reactivity without intracytoplasmic staining. Intracytoplasmic positivity for carcinoembryonic antigen in microvillous inclusion disease is explained by its presence in the glycocalyx within the microvillous inclusions. The demonstration of a distinct staining pattern for polyclonal carcinoembryonic antigen in routinely processed small bowel biopsy specimens provides a new useful criterion that complements other established techniques for accurate diagnosis of microvillous inclusion disease. PMID- 8244324 TI - Localization of p53 protein and human papillomavirus in anogenital squamous lesions: immunohistochemical and in situ hybridization studies in benign, dysplastic, and malignant epithelia. AB - p53 Protein is a 53-kd nuclear phosphoprotein believed to play an important role in controlling proliferation of neoplastic and normal cells. This "natural tumor suppressor" can be rendered ineffective (or oncogenic) by mutations in the p53 gene or by interactions with proteins synthesized by DNA-transforming viruses, including specific subtypes of human papillomavirus (HPV). We describe the localization of p53 protein in association with HPV in paraffin sections of a spectrum of benign, dysplastic, and malignant anogenital squamous epithelia using immunohistochemical and in situ hybridization techniques. p53 Was detected in 81% of the 48 cases studied. Immunoreactivity for p53 was seen in 83% of the benign and low-grade squamous intraepithelial lesions (SILs), in 73% of the high-grade SILs, and in 86% of the infiltrating squamous carcinomas. In high-grade SILs p53 staining was frequently observed in individual nuclei at various levels of the abnormal epithelium and in the basal layer of the adjacent epithelium, while in squamous metaplasia and low-grade SILs immunostaining for p53 was limited to the basal layer of the epithelium. p53 Was detected in a slightly higher percentage of HPV-positive than HPV-negative epithelia as determined by in situ hybridization. No correlation was observed between p53 immunoreactivity and HPV subtypes. p53 Protein and HPV were detected in anal lesions from a small group of human immunodeficiency virus-positive individuals. Antibodies currently available mainly demonstrate mutant forms of p53 protein that are associated with longer half-lives than the wild-type protein, but demonstration of p53 protein overexpression is not necessarily indicative of malignancy. PMID- 8244325 TI - Stereomicroscopic examination of surface morphology in colorectal epithelial tumors. AB - The mucosal surface structure of epithelial neoplasms in the large intestine was examined by a stereomicroscope, and the correlation between histologic and macroscopic features was studied. The surface structure of the colonic mucosa could be readily and clearly observed using the stereomicroscope after superficial staining, and neoplastic lesions were identified as well-defined foci of the unique structures that were conspicuously different from those in the normal mucosa. Five hundred two adenomas and 159 carcinomas were examined and divided into five types according to stereomicroscopic appearance: long ellipsoid (359 lesions), cerebriform (141 lesions), leaf-like (16 lesions), dense tiny pits (89 lesions), and devastated (56 lesions). In the first three types benign adenomas predominated in the majority of cases (94.7%, 72.3%, and 68.8%, respectively) and the variation in the surface features correlated well with the histologic types. In the latter two types the frequency of malignancy was strikingly high (49.4% and 92.9%, respectively), with the devastated mucosal appearance being highly indicative of invasive carcinoma. Macroscopically, exophytic polypoid configurations were frequently found in the first three types, whereas flat nonpolypoid configurations predominated in the cases with dense tiny pits. Our results indicate that there is a close relationship between the surface structure of epithelial neoplasms in the large intestine and their histologic features and growth patterns. PMID- 8244326 TI - Small lymphocytic non-Hodgkin's lymphoma of suppressor/cytotoxic T-cell phenotype [CD4(-), CD8(+)]. AB - An unusual case of small lymphocytic non-Hodgkin's lymphoma of suppressor/cytotoxic T-cell phenotype is described. The patient presented with significant involvement of the spleen, lymph nodes, and bone marrow, but without a leukemic phase. Most small lymphocytic lymphomas are of B-cell phenotype, and those of T-cell origin are overwhelmingly of T-helper/inducer phenotype. Furthermore, T-cell lymphoproliferative lesions of suppressor/cytotoxic phenotype generally present as leukemias comprising large granular lymphocytes. This case reveals that suppressor/cytotoxic T lymphocytes may give rise to a lymphoproliferative disorder with a distinctive phenotype and presentation. PMID- 8244327 TI - Glioblastoma multiforme of donor origin after renal transplantation: report of a case. AB - A case of transmission of a glioblastoma multiforme from the donor to a kidney transplant recipient in the absence of previous ventriculosystemic shunt is described. The recipient was a 48-year-old woman who developed a fever with no other associated symptoms 17 months posttransplant. Physical examination revealed a large nonpulsatile mass on the upper graft pole. Histopathologic examination showed a highly cellular neoplasm with fusiform and globoid cells, a high grade of nuclear pleomorphism and mitosis, necrosis with pseudopalisading, and vascular proliferation. Immunohistochemistry of the cells showed them to contain glial fibrillary acidic and S-100 proteins, consistent with a glioblastoma multiforme. We conclude that the risk of tumor transmission from donors with primary central nervous system tumors to kidney transplant recipients, is real and should be considered when evaluating a graft mass in such patients. PMID- 8244328 TI - Epithelioid leiomyoma of the breast with granular cell change: a case report. AB - A 42-year-old woman with an epithelioid granular cell leiomyoma of the right breast is reported. The cells were spindle to polygonal and displayed finely granular cytoplasm. The smooth muscle differentiation of this tumor was shown by the immunohistochemical positivity of the neoplastic cells with anti-alpha smooth actin and antidesmin antisera. Microfilaments with focal densities were present in the cytoplasm at an ultrastructural level. The granular cytoplasmic changes are related to a relevant number of lysosomes within the neoplastic cells. PMID- 8244329 TI - Liposclerosing myxofibrous tumor of bone. PMID- 8244330 TI - Papillary endothelial hyperplasia. PMID- 8244331 TI - Data on the CGG repeat at the fragile X site in the non-retarded Japanese population and family suggest the presence of a subgroup of normal alleles predisposing to mutate. AB - The fragile X mutation is the result of amplification in the repeat number of p(CGG)n in FMR-1; alleles with more than 52 repeats have been shown to be so unstable as to mutate in the repeat number in almost every transmission. To improve our understanding of mutations in normal alleles of FMR-1, the following studies were carried out in the Japanese population: a study on length variation in the repeat to determine the allele distribution of the repeat length in a non retarded population, family studies to observe new mutations in normal allele, and haplotype analyses with microsatellite markers flanking the repeat to confirm estimated mutation rates and founder chromosomes in the fragile X syndrome. Analysis of the p(CGG)n in 370 unrelated males detected 24 distinct alleles with repeats of 18-44. A comparison with previously reported data suggests the presence of racial/ethnic differences in the allele distribution. No premutation allele was found in 824 unrelated X chromosomes examined by the polymerase chain reaction and Southern blot analysis. Family studies detected one new mutation in a total of 303 meioses. However, the mutation rate was not in accordance with the expected or observed heterozygosities in the population or with linkage disequilibrium observed between the repeat numbers and the haplotypes of the markers flanking the CGG. The haplotype in the chromosome in which the new mutation was found was the same as that frequently found in the Japanese fragile X chromosomes, and the variance in the CGG repeat number was wider in chromosomes with the haplotypes frequently found in the fragile X chromosome than in those with the other haplotypes. These observations suggest that a subgroup is present in normal alleles and that this subgroup is more liable to mutate than others. PMID- 8244332 TI - Molecular genetics of GM2-gangliosidosis AB variant: a novel mutation and expression in BHK cells. AB - The GM2 activator is a hexosaminidase A-specific glycolipid-binding protein required for the lysosomal degradation of ganglioside GM2. Genetic deficiency of GM2 activator leads to a neurological disorder, an atypical form of Tay-Sachs disease (GM2 gangliosidosis variant AB). Here, we describe a G506 to C transversion (Arg169 to Pro) in the mRNA of an infantile patient suffering from GM2-gangliosidosis variant AB. Using the polymerase chain reaction amplification and direct-sequencing technique, we found the patient to be homozygous for the mutation, whereas the parents were, as expected, heterozygous. BHK cells transfected with a construct of mutant cDNA gave no GM2 activator protein detectable by the Western blotting technique, whereas those transfected by a wild type cDNA construct showed a significant level of human GM2 activator protein. The substitution of proline for the normal Arg169 therefore appears to result in premature degradation of the mutant GM2 activator, either during the post translational processing steps or after reaching the lysosome. The basis for the phenotype of GM2 gangliosidosis variant AB may therefore be either inactivation of the physiological activator function by the point mutation or instability of the mutant protein. PMID- 8244333 TI - Molecular and cytogenetic analyses of autism in Taiwan. AB - Karyotypic and DNA analyses were both performed on 104 autistic children referred from Taichung Autism Education Academy and Tainan Autism Association in Taiwan. The frequency of fragile sites of the autistic patients did not differ significantly from that of the normal individuals. Of the 12 autistic children with chromosomal abnormalities, 8 had the fragile X, 2 had Down syndrome, and the remaining had other aneuploid constitutions. The results of this study illustrate the contribution of chromosomal abnormalities or variants to the pathogenesis of infantile autism. PMID- 8244334 TI - Molecular analysis of factor VII deficiency in Italy: a frequent mutation (FVII Lazio) in a repeated intronic region. AB - Molecular defects and polymorphic haplotypes of coagulation factor VII gene were studied in eight unrelated Italian subjects with factor VII deficiency, seven having the factor VII- variant, one the factor VIIR variant. An intron 7 mutation, which alters the consensus donor splice site sequence, was found in six subjects. The presence of the founder effect is suggested by their common geographical origin (a mountain area in the Lazio region) and by the identical polymorphic haplotype underlying the mutation. A different mutation, also located in the 5' monomer of the repeated intron 7 sequence, was found in the heterozygous condition in a subject from Northern Italy. New polymorphic alleles were detected in the repeated intron 7 region in subjects from Eastern Africa. Two missense mutations in codon 97 (Gly-->Cys, Gly-->Ser), the first found in the compound heterozygous condition with the frequent intron 7 mutation, suggest the presence of a hot spot mutation site in the second epidermal growth factor domain. Two neutral dimorphisms at codon 333Ser and 115His were detected, the last in linkage disequilibrium with the 353Arg/Gln polymorphism, and showing differences in frequency in the FVII deficient and control subjects. PMID- 8244335 TI - Supernumerary marker chromosomes in peripheral blood cells of hepatitis B virus chronic carriers. AB - Cytogenetic analysis of metaphase chromosome spreads from peripheral blood cells of hepatitis B virus (HBV) chronic carriers revealed supernumerary marker chromosomes in 2 of the 46 individuals tested. Both individuals are phenotypically normal oriental males, and exhibit mosaicism with a 46,XY/47,XY,+mar/48,XY,+2mar profile in one, and a 46,XY/47,XY,+mar profile in other. Based on the reported frequency of unidentified supernumerary chromosomes (1:2,500) in 377,357 amniocentesis samples, the frequency seen (1:23) in the population of HBV chronic carriers sampled here appears unusually high. The possibility of a role for HBV in the generation of marker chromosomes is discussed. PMID- 8244336 TI - Human malformations similar to those in the mouse mutation disorganization (Ds). AB - We report two patients with malformations similar to those seen in mice with the disorganization (Ds) mutation. The first case has a body wall defect, limb malformation, and hamartoma, while the second case has a partially duplicated foot, in addition to the other anomalies. We discuss the implications that recent advances regarding the genetic analysis of the mouse Ds locus have for the search for the human homologue. PMID- 8244337 TI - Frequency of glucose-6-phosphate dehydrogenase (G6PD) mutations in Chinese, Filipinos, and Laotians from Hawaii. AB - In a Hawaii Hereditary Anemia Screening Project, 4,984 participants were tested for glucose-6-phosphate dehydrogenase (G6PD) deficiency by a filter paper blood spot fluorescence test. Abnormal samples and suspected heterozygotes were checked by quantitative G6PD assay (normal 4.5 to 14 units/g Hb). G6PD was deficient (< 1.5 units/g Hb) in 188 of 2,155 males; 7 other males had low activity (1.5 to 2.8 units/g Hb). The gene frequency, estimated from males after excluding referred and related cases, was 0.037 for Chinese, 0.134 for Filipinos, and 0.203 for Laotians. Among 2,829 females tested, family data showed 111 females were obliged to be at least heterozygous, regardless of G6PD activity, and 43 others had low G6PD activity. Most heterozygotes probably remained undetected by G6PD screening. In 28 females, activity was under 10%; in another 9 females, activity was < 1.5 units/g Hb. Since only 25 homozygotes would be predicted, this apparent excess of females with deficient activity could be due to unequal X-inactivation in some heterozygotes. DNA analysis by polymerase chain reaction amplification and special analytic procedures revealed 10 different missense mutations in 75 males. The nucleotide 835 A-->T and 1360 C-->T transitions were first detected in this Hawaiian Project; we found that the nucleotide 1360 mutation was the most common cause of G6PD deficiency in Filipinos. This is the first report of G6PD screening and analysis of molecular G6PD mutations in Filipino and Laotian populations. PMID- 8244338 TI - Diverse polymorphism within a short coding region of the human aldehyde dehydrogenase-5 (ALDH5) gene. AB - Human aldehyde dehydrogenase-5 gene (originally named as ALDHX) is expressed in liver and testis. The ALDH5 does not contain introns in the coding sequence for 517 amino acid residues. Within a short nucleotide region of the gene, the following three nucleotide changes were found in high frequencies, i.e., a silent C<-->T at nucleotide (nt) 183, C<-->T at nt 257 associated with a Val<-->Ala substitution, and T<-->G at nt 320 associated with a Arg<-->Leu substitution. The frequency of C at nt 183 is 81% in Caucasians and 65% in Japanese, and the difference is statistically not significant. The frequency of C at nt 257 is 76% in Caucasians and 55% in Japanese, and the difference is statistically significant (P = 0.02). The frequency of T at nt 320 is 71% in Caucasians, while it is only 27% in Japanese. The racial difference at nt 320 is highly significant (P < 0.001). No significant difference was found in the genotypes of the three nucleotide positions between alcoholic and nonalcoholic Caucasians within the limited numbers of subjects examined. PMID- 8244339 TI - Paternal duplication of chromosome 5q11.2-5q14 in a male born with craniostenosis, ear tags, kidney dysplasia and several other anomalies. AB - A de novo duplication of the proximal part of the long arms of chromosome 5 was found in a male born with craniostenosis, ear tags and kidney dysplasia. The nature of the chromosomal aberration was defined by fluorescence in situ hybridization and the origin of the duplication was traced by polymorphic DNA markers. A comparison is made with the published cases showing similar duplications in the long arm of chromosome 5. PMID- 8244340 TI - Hereditary triosephosphate isomerase (TPI) deficiency: two severely affected brothers one with and one without neurological symptoms. AB - A 13-year-old Hungarian boy (B.J.Jr.) with congenital haemolytic anaemia (CHA) and hyperkinetic torsion dyskinesia was found to have severe triose-phosphate isomerase (TPI) deficiency. One of his two brothers (A.J.), a 23-year-old amateur wrestler, has CHA as well, but no neurological symptoms. Both have less than 10% TPI activity and a highly increased dihydroxyacetone phosphate (DHAP) level in their red blood cells. Their TPI had a slow electrophoretic mobility and was heat unstable. Both parents and a third brother are healthy heterozygous carriers of the defect. A.J. represents a unique phenotype from the point of view that all published "homozygotes" had severe neurological alterations from infancy or early childhood except one infant who died at 11 months, probably too young for neurological symptoms to be noted. In contrast to the two affected Hungarian brothers all but one "homozygote" has died before the age of 6 years. The striking difference in the clinical course of the defect between the two brothers with the same severe red blood cell enzyme deficiency may originate from unusual differences between two double heterozygous brothers resulting inter alia in different levels of TPI expression in various tissues. Significantly lower TPI activities were found in both the T- and B-cells of the propositus as compared to the respective cells of the neurologically symptom-free brother. PMID- 8244341 TI - Spondyloepiphyseal dysplasia and precocious osteoarthritis in a family with an Arg75-->Cys mutation in the procollagen type II gene (COL2A1). AB - Direct sequencing of polymerase chain reaction (PCR)-amplified genomic DNA from a patient with spondyloepiphyseal dysplasia and precocious osteoarthritis revealed a single-base change in exon 11 of the type II procollagen gene (COL2A1), which produces an Arg-->Cys mutation in one allele. The proband is a member of a large Chilean kindred presenting with chondrodysplasia of the hips, knees, shoulders, elbows, and spine associated with severe, early-onset osteoarthritis. All affected individuals exhibit mildly short stature; in addition, five out of seven affected family members display shortened metacarpals or metatarsals. DNA from affected and unaffected family members was PCR-amplified and analysis of restriction digests of the products determined that the mutation segregated with the disease with a lod score of 2.2 at zero recombination. The mutation, which resides in the triple-helical region of type II procollagen at amino acid position 75, is the second example of an Arg-->Cys mutation in the COL2A1 gene in heritable cartilaginous disease and is the first example of a point mutation in the amino terminal region of the alpha 1(II) chain, that results in a spondyloepiphyseal dysplastic phenotype. PMID- 8244342 TI - Acceptor splice site mutation in the invariant AG of intron 5 of the protein C gene, causing type I protein C deficiency. AB - An acceptor splice-site mutation (3318, A-->G) in the invariant AG of intron 5 of the human protein C gene has been identified in a Spanish family with heterozygous type I protein C (PC) deficiency and thromboembolic disease. Family studies confirmed cosegregation of the mutation with type I PC deficiency. Computer analysis of the mutated sequence predicted the normal splicing site to be abolished by this mutation, whereas a cryptic splice site located two nucleotides downstream, in exon 6, is probably activated. According to this, 3318, A-->G should result in a frameshift with a stop at codon 119, in agreement with the presence of a type I or quantitative PC deficient phenotype in the affected members of the family. PMID- 8244343 TI - A rare reciprocal translocation (12;21) segregating for nine generations. AB - An autosomal reciprocal translocation (12;21) was found in five seemingly unrelated families in Finland. Three families had had multiple spontaneous abortions and two families had a child with Down's syndrome. The genealogies of the five families were traced using population registries, and four families were found to have a common ancestor born in 1752. Kinship to the fifth family could not be established but its ancestors were traced back to the same rural parishes as those of the four other families. The translocation segregated at the same frequency as normal chromosomes. A statistically insignificant increase in spontaneous abortions was detected when the matings of translocation carriers were compared with non-carrier matings. The increase may however be clinically significant. These results permit more accurate counselling in these and similar translocation families. PMID- 8244344 TI - Gaucher disease (Norrbottnian type III): probable founders identified by genealogical and molecular studies. AB - Gaucher disease type III (GD) is found at a high frequency in northern Sweden. The contemporary Swedish index families are found in two geographically distinct clusters with the highest world-wide frequency of type III GD. A single T-to-C transition in exon 10 has previously been identified in patients from one of the two isolates and we report there the same mutation in the second isolate. Mutational analysis was combined with a genealogical reconstruction of 19 contemporary index families. Both clusters were traced back to two corresponding pairs of ancestors over a 9-13 generation span. Molecular studies show that the two clusters are compatible with a single founder who arrived in northern Sweden in or before the 16th century. PMID- 8244345 TI - Assignment of the human aggrecan gene AGC1 to 15q25-->q26.2 by in situ hybridization. AB - The human aggrecan gene (AGC1) has been localized to 15q25-->q26.2 by in situ hybridization. Although no genetic diseases of connective tissue map to this location, the malignant melanoma-associated surface antigen mel-CSPG is located here; mel-CSPG is a chondroitin sulfate proteoglycan. This raises the possibility that AGC1 and mel-CSPG may be the same gene. PMID- 8244346 TI - A novel Cys-214-Ser mutation in the peripherin/RDS gene in a Japanese family with autosomal dominant retinitis pigmentosa. AB - We have screened for possible disease-causing mutations in the peripherin/retinal degeneration slow (RDS) gene in 13 Japanese families with autosomal dominant retinitis pigmentosa (ADRP). Using polymerase chain reaction-single strand conformation polymorphism analysis, a novel mutation at codon 214 was found in which the highly conserved cysteine was replaced with a serine in one family. The mutation at codon 214 was found in all three affected siblings of this family, but none of the 40 normal control individuals had this mutation. These results strongly suggest that the mutation is pathogenic for RP in this family. The clinical phenotype for this family is a late-onset form of ADRP. PMID- 8244347 TI - Centromeric association of a microchromosome in a Turner syndrome patient with a pseudodicentric Y. AB - A 12-year-old patient with Turner syndrome was found to have a complex mosaicism for a microchromosome (MC) and a psu dic(Y)(q11). The MC was smaller than Yp, appeared pale in G, C and late replicating bands, had a pair of small centromeric dots, was associated with other chromosomes in most metaphases, and was rather stable both in size and during mitosis. The psu dic(Y) was Cd-positive only at the active centromere, had two pericentromeric heterochromatic regions, and lacked the Yq12 band. No cells with both abnormal chromosomes were found. To evaluate the association of the MC with all ordinary chromosomes, 857 G-banded cells with the marker were screened. The MC was considered as "associated" whenever the distance between it and other chromosome(s) was equal to, or smaller than, 18p. Out of 848 associations registered, 489 (57.7%) were centromeric, 202 (23.8%) telomeric, and 157 (18.5%) interstitial; i.e., centromeric associations were overrepresented (P < 0.001) and showed a random distribution, except for an excessive involvement of chromosome 8. This association pattern, also exhibited by two similar MCs in human beings, the minute Y of a marsupial and certain B chromosomes in plants, probably reflects the Rabl orientation of chromosomes in interphase. PMID- 8244349 TI - Sexually transmitted diseases in modern China: a historical survey. AB - This paper points to the congruence between political and social variables and the epidemiology of sexually transmitted diseases (STDs) in modern China. STDs became a major health problem after the fall of the empire in 1911 and were only reluctantly addressed by a weak nationalist government during the 1930s. During the 1950s and 60s, the communist regime brought STDs under control, but problems have reappeared since reforms were implemented during the 1980s. Cultural values and social attitudes have also structured medical responses to venereal disease. From the reform movements between the two World Wars to the more recent communist health campaigns, medical theory has often been confused with moral prescription. PMID- 8244348 TI - The pathogenesis of the neurological complications of HIV-1 infection. PMID- 8244350 TI - Diagnosis of genital human papillomavirus (HPV) lesions in the male: correlation of peniscopy, histology and in situ hybridisation. AB - OBJECTIVE: To assess the diagnostic criteria of genital HPV lesions in male sexual partners of HPV infected women. METHODS: Peniscopically directed biopsy specimens (from 693 lesions in 300 men) were examined on light microscopy and in situ hybridisation (ISH) for HPV types 6,11,16,18,31,33 and 42. The predictive value of different histological criteria for ISH positivity was also evaluated using stepwise logistic regression analysis. RESULTS: Flat HPV lesions were most accurately predicted by the punctuation pattern on peniscopy, giving the concordance between peniscopy and histology between peniscopy and histology of 79.5% (66/83) and that between peniscopy and ISH of 56.6% (47/83). Diffuse acetowhite pattern disclosed a typical HPV lesion in only 17.8% (13/73), and HPV DNA was found in 11.0% (8/73) of cases. Of the 114 biopsy specimens from peniscopically healthy areas adjacent (0.5-1 cm) to the lesions, 93.0% (106/114) were normal on light microscopy, and HPV DNA was found in only 2.6%. Penile intraepithelial neoplasia (PIN) lesions were most frequently ISH positive, 81.1% (30/37), 50% showing HPV 16 and/or 18 DNA. Lesions classified as HPV-suspicious or nonspecific on light microscopy were HPV DNA-positive in 16.9% (11/65) and 8.1% (13/160), the frequency of high-risk HPV types being 3.1% and 1.3%, respectively. In logistic regression analysis, koilocytosis was the most powerful predictor of ISH-positivity in the flat lesions (without PIN), the risk ratio being 3.7. CONCLUSION: No conclusive peniscopic criteria for male HPV infections could be established, making histological evaluation mandatory. Care should be exercised in interpreting as HPV lesions the cases devoid of koilocytosis, HPV typing being essential in confirming the diagnosis in doubtful cases. PMID- 8244351 TI - HIV and other sexually transmitted diseases at a rural hospital in Zimbabwe. AB - OBJECTIVE: To define the epidemiological characteristics of STD patients attending an outpatient clinic in rural Zimbabwe, to examine the aetiologic agents causing infection and to determine their relationship with HIV infection. SUBJECTS: 319 men and 146 women, making a sample of about 7% all patients attending an STD clinic during the 3 month study period. Microbiological data were collected from 104 men and 72 women selected randomly from these. Pregnant women were excluded and patients who had received antibiotics within the previous 14 days were excluded from the microbiology sub-sample. SETTING: An outpatient STD clinic at a District Hospital on a major truck route about 300 km north of the capital, Harare. METHODS: All new patients attending the clinic during a 3 month period were enrolled for clinical and epidemiological investigations using a standard procedure. Specimens for microbiological investigation were taken from every second patient seen on the first three days of each week. RESULTS: The typical patient was male (m:f ratio 2.2) aged 20-29 years (68% patients), not married (56% men) and in paid employment (66% men vs. 27% for the district). In men the most common presenting feature was genital ulceration, while in women, discharges were more common. Genital warts were noted frequently in both sexes. In the sub-sample examined microbiologically, H ducreyi was isolated from 46% ulcers clinically diagnosed as chancroid, and motile spirochaetes were detected in 25% painless ulcers. Neither of these were detected in ulcers in women, but HSV antigen was found as frequently in ulcers from men (19%) as from women (17%). In patients with genital discharges, gonococcal infection occurred in 64% men and 17% women, while T vaginalis was isolated from 39% women and only 8% men. Over 60% gonococcal isolates were PPNG, and 18% showed in vitro resistance to tetracycline. Yeasts, mainly C albicans were isolated from 42% women with a discharge and 25% women with ulcers. In men the presence of yeasts was associated with superficial ulceration and itchiness of the glans. Positive HIV-1 serology was found in 64% patients. There was no statistical association with current genital ulcers, though there was an association with previous STD episodes and particularly with serological evidence of syphilis. Apart from yeasts, there was no association between positive HIV-1 serology and the presence of pathogens in the genital tract. CONCLUSIONS: The high prevalence of HIV-1 antibodies in STD patients in Karoi suggests integration of STD and AIDS control programmes to be a necessity. Since paid employment was a common feature of both STD clinic attendance and HIV-1 seropositivity, these programmes may be effectively directed through the work place. PMID- 8244352 TI - The association between sexually transmitted pathogens and cervical intra epithelial neoplasia in a developing community. AB - OBJECTIVE--To determine the association of sexually transmitted pathogens in women with cervical intra-epithelial neoplasia (CIN). SETTING--An urban tertiary referral hospital serving a large indigent developing community. PARTICIPANTS--48 women attending a colposcopy clinic and 49 women attending a family planning clinic. METHODS--Vaginal, endocervical, rectal swab specimens and sera were collected for the detection of sexually transmitted pathogens. Cervical cytology was performed on all patients. Women attending the colposcopy clinic had confirmation of abnormal cervical cytology by colposcopic directed biopsy. RESULTS--The mean age of women with CIN (33 years) was significantly greater than that of the women without CIN (28 years) and that of the family planning group (26 years). There was a high prevalence of sexually transmitted pathogens in all women. A significantly higher prevalence of bacterial vaginosis was found in women with CIN compared to those without (50% vs 20%; p = 0.034). The human papilloma virus (HPV) was detected in 46% of women with CIN and 65% of those without CIN. Chlamydia trachomatis (21%) and Trichomonas vaginalis (39%) were detected frequently in women with CIN. C. trachomatis (14%-21%) was detected more frequently than Neisseria gonorrhoeae (3-5%) in all asymptomatic women studied. CONCLUSION--This study demonstrates a high prevalence of sexually transmitted pathogens in women with and without CIN as well as family planning clinic attenders. Bacterial vaginosis was a significant finding in women with CIN. C. trachomatis was detected in a high proportion of all women studied and found more commonly than N. gonorrhoeae. We therefore recommend that all women attending gynaecological services in a developing community be investigated and treated for sexually transmitted diseases. PMID- 8244353 TI - Microbiology of acute epididymitis in a developing community. AB - OBJECTIVE: To investigate the aetiology of acute epididymitis in a developing community with a view of determining appropriate antimicrobial therapy. SETTING: City Health Sexually Transmitted Diseases Clinic, King Edward VIII Hospital, Durban, South Africa. PARTICIPANTS: 144 adult men with clinically diagnosed acute epididymitis. METHOD: Endourethral swab and midstream urine (MSU) specimens were processed to detect sexually transmitted pathogens and urinary tract infections. RESULTS: The majority of patients (93%) were less than 35 years of age. Neisseria gonorrhoeae and/or Chlamydia trachomatis were detected in 78% of patients: N gonorrhoeae in 57%, C trachomatis in 34% and both in 13%. Escherichia coli was cultured more frequently from MSU specimens of older patients, 30% versus 3%. In 53% of patients urethritis was diagnosed by the presence of inflammatory cells in endourethral smears in the absence of a visible urethral discharge. CONCLUSION: In our setting of a busy clinic with limited facilities, we recommend the performance of a Gram stain on endourethral specimens from patients with acute epididymitis. If inflammatory cells and Gram negative diplococci are detected, treatment with antimicrobial agents to cover both penicillinase-producing N gonorrhoeae strains and C trachomatis is recommended. If Gram negative diplococci are not detected in the presence of microscopic evidence of urethritis, treatment for chlamydial infection alone is recommended. PMID- 8244354 TI - The incidence of gonorrhoea and the antibiotic sensitivity of gonococci in Australia, 1981-1991. The Australian Gonococcal Surveillance Programme. AB - OBJECTIVE: To review and analyse the changing incidence of gonorrhoea and the increasing antibiotic resistance in gonococci in Australia from 1981 to 1991. DESIGN: Use of data from the sample of gonorrhoea in Australia examined by the Australian Gonococcal Surveillance Programme (AGSP), a continuing long-term multi centric study of gonococcal disease and gonococcal susceptibility to antibiotics, over the period 1 July, 1981 to 30 June, 1991. RESULTS: Over 32,000 cases and strains from defined sources were examined in the 10 year study period. The number of cases of gonorrhoea in the sample decline from a peak of 6599 in 1982 1983 to 1121 in the final year under review, a reduction of 83%. Periods when greater than average reductions in incidence occurred in different groups were noted. Ano-rectal gonorrhoea in men decreased sharply in 1985 during an overall decline of 92.5% recorded between 1 July, 1981 to 30 June, 1987. However, the incidence of ano-rectal cases in males rose in subsequent years while gonorrhoea, overall, continued to decrease and at a greater rate after 1985. Antibiotic resistance in gonococci in Australia was manifested both as a progressive increase in the levels of intrinsic resistance to the penicillins and through the appearance and spread of penicillinase-producing N gonorrhoeae (PPNG). At the end of the review period in June, 1991, 8.8% of gonococcal isolates in Australia showed high levels of intrinsic resistance to the penicillins and 13% of strains were PPNG. These separate mechanisms of resistance appeared at different times in different parts of Australia, and their importance also varied throughout the country. Most infections with PPNG were acquired by men overseas whereas most women with PPNG were infected locally. Endemic spread of PPNG was a significant problem in Sydney and Melbourne, but decreased in importance in the later years of the study. CONCLUSIONS: In the past decade a large reduction in the incidence of gonorrhoea and, by implication, other STDs has occurred in the past decade in Australia. In some groups of patients the decline in incidence is continuing while in others a slight increase has been noted. Resistance to antibiotics of gonococci in Australia was mainly restricted to the penicillins, but through both chromosomal and plasmid-mediated mechanisms. This resistance was seen particularly in Sydney and Melbourne where endemic cycles of transmission of PPNG were established, and in infected travellers from S-E Asia in other centres. Valid and comparable, and regionally relevant data are a continuing requirement for assessing and modifying antibiotic treatment regimens for gonococcal disease. PMID- 8244355 TI - Outpatient follow-up in women with HIV infection in Parkside Health Authority (UK). AB - OBJECTIVE: To describe patterns of attendance for follow-up among HIV infected women in Parkside, UK and their correlates. DESIGN: Retrospective cohort study. SUBJECTS: 103 HIV infected women. MAIN OUTCOME MEASURES: Whether patients attended for follow-up between three and 18 months. RESULTS: 31% of women were married and 46% had children. Women born in sub-Saharan Africa were significantly less likely to attend for follow-up after three months (56%) than women born in other areas who had acquired HIV either heterosexually (82%) or through injecting drug use (81%). This pattern persisted on multivariate analysis controlling for whether women were symptomatic, had had a previous positive test, were married or had children. CONCLUSIONS: HIV positive sub-Saharan African women are less likely to reattend for follow-up than women with heterosexually acquired HIV from other areas or those who acquired infection through intravenous drug use. Further studies are needed to identify barriers to follow-up for women and to shape the development of more appropriate and accessible services for HIV infected women, especially those of sub-Saharan African origin. PMID- 8244356 TI - Defective IgG2 response to Pneumovax in HIV seropositive patients. AB - OBJECTIVE: To determine whether HIV (Human Immunodeficiency Virus) antibody positive adults are capable of mounting an effective immune response when immunized with polyvalent pneumococcal vaccine. DESIGN: 28 patients (nine homosexual men, one bisexual man, three heterosexual females, 10 injecting drug abusers, five haemophiliacs) and 11 healthy volunteers, were immunised with Pneumovax II and titres of IgG1 and IgG2 specific antibody measured before and 1 month after immunisation. Magnitude of immune response was related to CD4 T lymphocyte count at time of immunisation to establish whether responses are better in early disease. MAIN OUTCOME MEASURES: Based on our data in healthy volunteers we defined an adequate response to Pneumovax II as a post immunisation IgG2 antibody level at least 50% greater than the pre immunisation level. RESULTS: The magnitude of the response was significantly higher in the normal volunteers (U = 95; p = 0.0328). Adequate IgG2 responses were seen in 11/11 normals but in only 14/28 HIV seropositives (chi 2 = 8.58; p < 0.01). Poor responses were unrelated to the CD4 T-lymphocyte count at immunisation. Absolute IgG2 deficiency accounted for the poor response in only 1 HIV patient. CONCLUSION: 50% of HIV antibody seropositive individuals fail to mount adequate immune responses to polyvalent pneumococcal vaccine. Non responders are unlikely to be protected. PMID- 8244357 TI - Attitudes of medical students to HIV and AIDS. AB - OBJECTIVE--To assess the knowledge and attitudes of medical students to HIV/AIDS and whether attitudes correlate with knowledge and clinical experience. To determine if students felt adequately prepared to deal with medical and psychological aspects of HIV/AIDS. SUBJECTS AND METHODS--The subjects consisted of 190 London and 99 Cambridge medical students at the end of their genitourinary medicine attachment, plus 230 Cambridge medical students at the end of their second pre-clinical year. Between March 1991 and February 1992 all were asked to complete an anonymous questionnaire, covering factual knowledge and attitudes towards HIV/AIDS. MAIN RESULTS--Cambridge genitourinary medicine students, despite spending less time studying HIV infection than their London counterparts gave more correct answers to the factual questions, although this difference did not reach significance (52.4% vs. 47.5%, p = 0.14). One third of students believed that many health care workers were at high risk of acquiring HIV at work and one fifth thought doctors should have the right to refuse to treat people with HIV. Fourteen percent of Cambridge genitourinary medicine students indicated that most British people with HIV have only themselves to blame, by comparison with 4% of London students (p = 0.003). Thirty-nine per cent of Cambridge genitourinary medicine students expressed reluctance to care for someone with AIDS by comparison with 10% of London students (p = 0.0001). CONCLUSIONS--It is important that medical educators convey accurate information about HIV, including the actual risks posed by occupational exposure and try to ensure that medical students spend sufficient time seeing patients with HIV/AIDS during their training. PMID- 8244358 TI - The epidemiology of herpes simplex types 1 and 2 infection of the genital tract in Edinburgh 1978-1991. AB - INTRODUCTION: The changing epidemiology of genital herpes in Edinburgh is described in relation to herpes simplex virus (HSV) Type 1 and herpes simplex virus Type 2 infection over a period of 14 years. METHODS: 2018 episodes of genital herpes in 1794 patients over a 14 year period were assessed. Data on age, sex, sexual orientation, geographical origin and herpes antibodies were also analysed. RESULTS: The proportion of cases that were HSV Type 1 increased over the period from approximately 20% to over 40%. Type 1 infection is more common in the young, in women and as a primary infection. CONCLUSIONS: HSV Type 1 is of increasing importance as a cause of genital herpes in our population. This may reflect changes in sexual attitudes and practises over the past decade. PMID- 8244359 TI - Adjustment to the psychological and social sequelae of recurrent genital herpes simplex infection. AB - OBJECTIVE: To investigate whether adverse psychological consequences and impaired sexual and interpersonal functioning are present in individuals suffering from recurrent genital herpes simplex infection (HSV). DESIGN: A questionnaire-based study completed by subjects defined as having more than one episode of HSV infection. SUBJECTS: Completed questionnaires were returned from 90 subjects; 40 from the department of genitourinary medicine at our hospital and 50 from members of the Herpes Association. RESULTS: Stress and being physically run-down were identified as provoking factors by the majority of individuals (78% and 56% respectively). The frequency of sexual activity was not different when comparing rates before and after infection (p < 0.001) and the majority of subjects had told partners of their infection. Women reported significantly greater disturbances in several psychological variables and reported a greater decrement in their general health. However, overall there were no differences in the psychological sequelae following herpes infection. CONCLUSIONS: The study suggests that, given time, most people are able to adjust psychologically to having recurrent herpes infection. However, for a minority of subjects this is not the case and these individuals may require psychotherapeutic intervention. PMID- 8244360 TI - Effect of lactic acid suppositories compared with oral metronidazole and placebo in bacterial vaginosis: a randomised clinical trial. AB - OBJECTIVE: To compare the effect of lactic acid locally, metronidazole orally and placebo in women with bacterial vaginosis. DESIGN: Randomised clinical trial. SETTING: 30 general practices in the Netherlands. PATIENTS: 125 women consulting the general practitioner for symptomatic bacterial vaginosis. MAIN OUTCOME MEASURES: Duration of subjective symptoms, recurrence of symptoms, clinically diagnosed cure, adverse events. RESULTS: Survival analysis showed a significantly faster disappearance of symptoms in the metronidazole category compared with both lactic acid and placebo (p = 0.0005 metronidazole v placebo, p = 0.0002 metronidazole v lactic acid p = 0.6521 lactic acid v placebo [The stratified Mantel Cox test]). The median duration until absence of symptoms was 21 days for metronidazole and 80 days for placebo. Disappearance of symptoms did not occur in 50% of the lactic acid group in 90 days. Recurrence rates of symptoms were similar over the treatment categories (p = 0.13 metronidazole v placebo and p = 0.12 lactic acid v placebo). After 2 weeks cure rates (cure defined as less than three of four clinical criteria present) were 83%, 49% and 47% for metronidazole, lactic acid and placebo category respectively. At that time cure rates (cure defined as none of three clinical criteria present) were 10%, 0% and 3%. After four weeks and three months these figures were: 55%, 20%, 20% and 64%, 28%, 28%. No differences in adverse events were found between the three interventions. CONCLUSIONS: Lactic acid suppositories are ineffective, metronidazole capsules are effective on signs and symptoms in bacterial vaginosis. A considerable proportion of the patients recover without active medication. PMID- 8244361 TI - Factors associated with genital chlamydial and gonococcal infection in males. AB - BACKGROUND: Predictors of chlamydia and gonorrhoea can be used to increase the cost-effectiveness and acceptability of screening programmes, and allow targeting of control strategies. METHOD: All men attending an STD clinic in 1988-1990 were offered screening for chlamydia and gonorrhoea, and the test results correlated with a wide range of potential predictors using multiple logistic regression. RESULTS: Of 9622 attenders, 7992 (82.3%) were tested over a total of 10,110 episodes for chlamydia and 10,090 episodes for gonorrhoea, yielding 729 (7.2%) chlamydial and 123 (1.2%) gonococcal infections. Having urethral discharge and/or dysuria, being heterosexual, and STD contact, unmarried, uncircumcised, tattooed and not having had an STD previously were independently associated with chlamydial infection. Having urethral discharge and/or dysuria, being Aboriginal, an STD contact, homosexual, uncircumcised, tattooed and having sex outside the state in the past three months, no steady partner in the past three months and multiple partners in the past month were associated with urethral gonococcal infection. Selective screening criteria for gonorrhoea provided 90% of positives, eliminated the need for 58% of tests and resulted in an increased yield ratio of 2.2 whereas the corresponding outcomes for screening criteria for chlamydia were 93% 20% and 1.2 respectively. CONCLUSIONS: The widespread influence of confounding on potential predictors for both gonorrhoea and chlamydia may provide misleading indicators of risk factors by univariate analysis. In the setting studied the benefits of selective screening for gonorrhoea in men would be substantial, whereas satisfactory criteria for selective screening for chlamydia could not be identified. PMID- 8244362 TI - Cervical sampling for diagnosis of genital chlamydial infection with a new brush device. AB - OBJECTIVES: To compare a new sampling device, a brush, Accellone-Multi-Instrument (AMI), with a dacron-tipped swab for detection of Chlamydia trachomatis in endocervical specimens, and to evaluate if consecutive multiple cervical sampling as compared with such a single specimen would increase the sensitivity. METHODS: 501 females attending an STD clinic and 172 females attending a family planning clinic were examined prospectively. Two cervical specimens were collected from each woman. C trachomatis were detected by culture or enzyme immunoassay (IDEIA III). Positive EIA samples were confirmed by a direct immunofluorescent test. RESULTS: When cervical specimens were collected with the brush as the first device, 92% of the culture-positive cases were detected, and when the samples were collected with the dacron-tipped-swab first, 84% of the culture-positive cases were detected (p < 0.05). The first collected specimen detected 89% of the culture-positive cases and 81% of those that were positive by IDEIA. CONCLUSIONS: The study indicates that the AMI brush is superior to non-toxic, dacron-tipped swabs for detection of C trachomatis in cervical samples by cell culture but not by ELISA, and that the sensitivity could be improved by analysing multiple cervical samples. PMID- 8244363 TI - Clinical features and management of recurrent balanitis; association with atopy and genital washing. AB - OBJECTIVE: To evaluate clinical features and diagnostic investigations in patients with recurrent or unresponsive balanitis in order to institute rational management. DESIGN: Forty-three patients presenting to a genitourinary medicine clinic with recurrent or persistent balanitis were studied. All patients were asked whether they had a history of atopic illness and about their practice of genital washing. All patients were investigated by taking a swab specimen from the preputial area for bacterial and viral culture and 30 underwent biopsy of the affected skin. Follow-up was between three and six months. SETTING: Outpatient genitourinary medicine clinic, St Mary's Hospital, London, UK. RESULTS: In 31 (72%) of the patients a diagnosis of irritant dermatitis was made. In comparison with the remaining patients, they had a greater lifetime incidence of atopic illness and more frequent daily genital washing with soap. For 28 (90%) of these patients, use of emollient creams and restriction of soap washing alone controlled symptoms satisfactorily. For the remaining 12 patients, a variety of diagnoses were made. Biopsy proved a well tolerated and diagnostic investigation, but the isolation of microbial pathogens from preputial swabs was irrelevant to management. CONCLUSION: A history of atopic illness and of the practice of penile washing are important aspects in the evaluation of patients with recurrent balanitis. Biopsy is an important investigation in the condition when it does not seem to be caused by irritant dermatitis. PMID- 8244364 TI - Random amplification of polymorphic DNA of penicillinase-producing Neisseria gonorrhoeae strains. PMID- 8244365 TI - Direct culture of gonococci in the patient care area. PMID- 8244366 TI - Hepatitis B infection post Engerix B vaccination. PMID- 8244367 TI - Non tuberculous cavitory disease in a west African man with AIDS. PMID- 8244368 TI - The localisation of treponemes in skin biopsies. PMID- 8244369 TI - The localization of a gene causing X-linked cleft palate and ankyloglossia (CPX) in an Icelandic kindred is between DXS326 and DXYS1X. AB - The locus responsible for X-linked, nonsyndromic cleft palate and/or ankyloglossia (CPX) has previously been mapped to the proximal long arm of the human X chromosome between Xq21.31 and q21.33 in an Icelandic kindred. We have extended these studies by analyzing an additional 14 informative markers in the family as well as including several newly investigated family members. Recombination analysis indicates that the CPX locus is more proximal than previously thought, within the interval Xq21.1-q21.31. Two recombinants place DXYS1X as the distal flanking marker, while one recombinant defines DXS326 as the proximal flanking marker, an interval of less than 5 cM. Each of the flanking markers recombines with the CPX locus, giving 2-point lod scores of Zmax = 4.16 at theta = 0.08 (DXS326) and Zmax = 5.80 at theta = 0.06 (DXYS1X). PMID- 8244370 TI - Mapping of aldose reductase gene sequences to human chromosomes 1, 3, 7, 9, 11, and 13. AB - Aldose reductase (alditol:NAD(P)+ 1-oxidoreductase; EC 1.1.1.21) (AR) catalyzes the reduction of several aldehydes, including that of glucose, to the corresponding sugar alcohol. Using a complementary DNA clone encoding human AR, we mapped the gene sequences to human chromosomes 1, 3, 7, 9, 11, 13, 14, and 18 by somatic cell hybridization. By in situ hybridization analysis, sequences were localized to human chromosomes 1q32-q42, 3p12, 7q31-q35, 9q22, 11p14-p15, and 13q14-q21. As a putative functional AR gene has been mapped to chromosome 7 and a putative pseudogene to chromosome 3, the sequences on the other seven chromosomes may represent other active genes, non-aldose reductase homologous sequences, or pseudogenes. PMID- 8244371 TI - Organization of the murine T-cell receptor gamma locus. AB - Murine T cells carry either of two antigen receptors on their surface: alpha beta or gamma delta T-cell receptors (TCR). The function of gamma delta T cells is not well understood. In this population, subsets exist with tissue-specific localization, with invariant variable regions, and with specificity for self antigens. We have analyzed the murine TCR gamma locus by field inversion gel electrophoresis and cosmid cloning. The structure of the complete TCR gamma locus, the first complete physical map of any mouse TCR locus, is reported here. The locus spans 205 kb and consists of four clusters of genes and gene segments. Each cluster contains one or more variable elements, one joining element, and one constant element. We demonstrate that the C gamma 4 gene is distinguished from the other C gamma genes in that it has an additional hinge-encoding exon. New enhancer-like elements are identified in two of the clusters, downstream from the C gamma genes. The most upstream cluster has four V elements and resembles the human TCR gamma locus. The four clusters probably arose via duplications. PMID- 8244372 TI - Application of cDNA selection techniques to regions of the human MHC. AB - Identification of transcribed sequences by cDNA selection is a potentially rapid and efficient way of scanning large genomic DNA fragments for the presence of genes. To evaluate this approach further, we have applied it to three yeast artificial chromosomes (YACs) and examined the products obtained from a total of about 1100 kb from two regions of the human major histocompatibility complex (MHC). One YAC was derived from an extensively studied portion of the Class II region of the MHC. The cDNAs recovered from this YAC included representatives of the previously described genes as well as one or more cDNA clones not described in the databases. A second YAC spanned about 330 kb of DNA surrounding the Class I gene HLA-A. In addition to Class I clones, 10 distinct cDNA products were identified from this YAC. A third YAC contained about 700 kb of human DNA, including 260 kb of overlap with the second YAC, and recovered an additional cDNA complementary to YAC B30 H3 DNA. Overall, the method is shown to be able to detect very scarce cDNAs and to detect a large fraction of coding sequences in YAC clones. Advantages and limitations of the approach are discussed. PMID- 8244373 TI - Using mitotic recombinant mutant clonal lymphocytes for physical mapping of polymorphic loci on the short arm of human chromosome 6. AB - Immunoselection has been used to identify human lymphocytes that have undergone spontaneous mutation resulting in the loss of expression of one of the codominant HLA-A alleles. Approximately 35% of such mutations are the consequence of mitotic recombination events. Mitotic recombination is the result of nonsister chromatid exchange that leaves the mutated cell homozygous for all loci distal to the crossover point. The location of the crossover has been regionalized for 99 independently derived mutant lymphocyte clones by identification of their loss or retention of heterozygosity at seven reference polymorphic loci on chromosome 6. If a polymorphic locus of unknown map position is studied in clones from this ordered set of mitotic recombinants, and clones that display loss of heterozygosity and retention of heterozygosity of the locus are observed, then the map position of the locus is between the appropriate reference loci of the ordered set. The newly mapped locus becomes a reference locus in turn. In this way the mitotic recombinant mutant clones can be used to generate an ordered set of crossover points with a theoretical resolution limited only by the number of mutants generated. In this paper such a set of mutants is used to refine or confirm the map position of eight polymorphic loci on chromosome 6. PMID- 8244374 TI - A high-resolution linkage map of human 9q34.1. AB - A map comprising 16 distinct markers with heterozygosities of 0.61-0.92 for a 10 cM region of human 9q34.1 is presented. The map incorporates four genes and has a maximum intermarker interval of 2.1 cM. Markers were analyzed in the Venezuelan reference pedigrees and all were placed uniquely in the map with a minimum likelihood of 676:1. The map should prove useful in analysis of families segregating dystonia and tuberous sclerosis, as the DYT1 and TSC1 loci map within this region. PMID- 8244375 TI - The Ah receptor nuclear translocator gene (ARNT) is located on q21 of human chromosome 1 and on mouse chromosome 3 near Cf-3. AB - We have mapped the Ah receptor nuclear translocator (ARNT) gene to a conserved linkage group located on mouse chromosome 3 and human chromosome 1. EcoRI digested DNA from a panel of 17 human x mouse somatic cell hybrids was probed with a cDNA fragment of the human ARNT gene. Six of the 17 independent mouse x human hybrids were positive for human bands. Human chromosome 1 showed complete cosegregation with the gene, whereas discordant segregation was observed for all other human chromosomes. The human gene was localized to 1q21 by using DNA from mouse x human hybrid clones that retain translocations involving human chromosome 1, by segregation analysis in nine informative CEPH families, and by in situ hybridization. The mouse homologue was mapped to mouse chromosome 3 using a panel of 16 hamster x mouse somatic cell hybrids. Six of 16 mouse x hamster hybrids were positive for mouse bands, showing complete concordance with mouse chromosome 3. The mouse Arnt gene was regionally mapped on chromosome 3, using linkage analysis in an interspecific backcross. The results indicate that the mouse gene resides about 40 cM from the centromere and about 10 cM proximal to Cf-3, the gene for tissue factor. PMID- 8244376 TI - Conserved synteny between pig chromosome 8 and human chromosome 4 but rearranged and distorted linkage maps. AB - The porcine genes encoding interleukin 2, alcohol dehydrogenase (class I) gamma polypeptide, and osteopontin were mapped to chromosome 8 by linkage analysis. Together with previous assignments to this chromosome (the albumin, platelet derived growth factor receptor A, and fibrinogen genes), an extensive syntenic homology with human chromosome 4 was discovered. Loci from about three-quarters of the q arm of human chromosome 4 are on pig chromosome 8. However, the linear order of the markers is not identical in the two species, and there are several examples of interspecific differences in the recombination fractions between adjacent markers. The conserved synteny between man and the pig gives strong support to a previous suggestion that a synteny group present in the ancestor of mammalian species has been retained on human chromosome 4q. Since loci from this synteny group are found on two cattle chromosomes, the bovine rearrangement must have occurred after the split of Suidae and Bovidae within Artiodactyla. PMID- 8244377 TI - The oligodendrocyte-myelin glycoprotein of mouse: primary structure and gene structure. AB - The oligodendrocyte-myelin glycoprotein (OMgp), a phosphatidylinositol-linked membrane glycoprotein expressed in the brain, is in man encoded by a gene that is entirely within an intron of and on the strand opposite to the neurofibromatosis type 1 (NF1) gene. We obtained two distinct overlapping DNA clones from a mouse genomic library that contain the OMgp gene from mouse (mOMgp). There is a single intron in the 5' untranslated region in exactly the same position as the sole intron in the gene for the human OMgp (hOMgp). A repeat, (TC)24, in the mouse intron divides it into 5' and 3' segments that have 72 and 93% sequence identity, respectively, with the human gene. The deduced unprocessed polypeptides of both species have 440 amino acids and the similarity of the primary structures of mOMgp and hOMgp indicates conservation of function. The conservation of the nucleotide sequences of the coding, noncoding, and flanking regions of the two genes is remarkable and raises the possibility that the nucleotide sequence may serve a function that is separate from the role of encoding OMgp. PMID- 8244378 TI - High-density genetic map of the BRCA1 region of chromosome 17q12-q21. AB - To facilitate the positional cloning of the breast-ovarian cancer gene BRCA1, we constructed a high-density genetic map of the 8.3-cM interval between D17S250 and GIP on chromosome 17q12-q21. Markers were mapped by linkage in the CEPH and in extended kindreds in our breast cancer series. The map comprises 33 ordered polymorphisms, including 12 genes and 21 anonymous markers, yielding an average of one polymorphism every 250 kb. Twenty-five of the markers are PCR-based systems. The order of polymorphic genes and markers is cen-D17S250-D17S518-HER2 THRA1-RARA-D17S80 -KRT10-[D17S800-D17S857]-GAS- D17S856-EDH17B-D17S855-D17S859 D17S858-[++ +PPY-D17S78]-D17S183-EPB3-D17S579- D17S509-[D17S508-D17S190 = D17S810]-D17S791-[D17S181 = D17S806]-D17S797- HOX2B-GP3A-[D17S507 = GIP]-qter. BRCA1 lies in the middle of the interval, between THRA1 and D17S183. Markers from this map can be used to determine whether cancer is linked to BRCA1 in families, to evaluate whether tumors have lost heterozygosity at loci in the region, and to identify probes for characterizing chromosomal rearrangements from patients and from tumors. PMID- 8244379 TI - Multicolor FISH mapping with Alu-PCR-amplified YAC clone DNA determines the order of markers in the BRCA1 region on chromosome 17q12-q21. AB - A gene designated BRCA1, implicated in the susceptibility to early-onset familial breast cancer, has recently been localized to chromosome 17q12-q21. To date, the order of DNA markers mapped within this region has been based on genetic linkage analysis. We report the use of multicolor fluorescence in situ hybridization to establish a physically based map of five polymorphic DNA markers and 10 cloned genes spanning this region. Three cosmid clones and Alu-PCR-generated products derived from 12 yeast artificial chromosome clones representing each of these markers were used in two-color mapping experiments to determine an initial proximity of markers relative to each other on metaphase chromosomes. Interphase mapping was then employed to determine the order and orientation of closely spaced loci by direct visualization of fluorescent signals following hybridization of three probes, each detected in a different color. Statistical analysis of the combined data suggests that the order of markers in the BRCA1 region is cen-THRA1-TOP2-GAS-OF2-17HSD-248yg9-RNU 2-OF3-PPY/p131-EPB3-Mfd188- WNT3-HOX2-GP3A-tel. This map is consistent with that determined by radiation reduced hybrid mapping and will facilitate positional cloning strategies in efforts to isolate and characterize the BRCA1 gene. PMID- 8244380 TI - A radiation hybrid map of the BRCA1 region of chromosome 17q12-q21. AB - The chromosomal region 17q12-q21 contains a gene (BRCA1) conferring susceptibility to early-onset familial breast and ovarian cancer. An 8000-rad radiation-reduced hybrid (RH) panel was constructed to provide a resource for long-range mapping of this region. A large fraction of the hybrids (approximately 90%) retained detectable human chromosome 17 sequences. The complete panel of 76 hybrids was scored for the presence or absence of 22 markers from this chromosomal region, including 14 cloned genes, seven microsatellite repeats, and one anonymous DNA segment. Statistical analysis of the marker retention data employing multipoint methods provided both comprehensive and framework maps of this chromosomal region, including distance estimates between adjacent markers. The comprehensive RH map includes 17 loci and spans 179 cRays(8000). Likelihood ratios of at least 1000:1 support the 10-locus framework order: cen-D17S250-ERBB2 (THRA1, TOP2A)-D17S855-PPY-D17S190-MTBT1-GP3A++ +-BTR-D17S588-tel. The order obtained from RH mapping, when used in conjunction with other methods, will be useful in linkage analysis of breast cancer families and will facilitate the development of a physical map of this region. PMID- 8244381 TI - Ordered shotgun sequencing, a strategy for integrated mapping and sequencing of YAC clones. AB - Ordered shotgun sequencing proposes to organize the mapping and sequencing of YACs with a hierarchical strategy that incorporates a feedback loop. Building on current protocols, a YAC is subcloned into plasmids, plasmid insert ends are sequenced, and the sequences are overlapped to create a partial map. Complete sequencing then starts with plasmids whose end-sequence tracts have overlapped, but to a minimal extent. The next plasmids to be sequenced are again selected for least overlap, striking out progressively to span the YAC with minimal directed gap-filling. Simulations support its feasibility and indicate that during the generation of the complete sequence, the approach facilitates the early choice of regions for selective sequencing, for example, for coding units. The sequencing of plasmids would also require less redundancy, and discriminate repetitive sequences more easily, than random sequencing across larger clones. The overall effort scales with YAC size and can be further reduced by additional mapping information. PMID- 8244382 TI - cDNA cloning and expression of human glutamyl aminopeptidase (aminopeptidase A). AB - The murine B-lymphocyte differentiation antigen BP-1/6C3 has been identified as glutamyl aminopeptidase (E-AP), formerly known as aminopeptidase A, the new gene symbol for which is ENPEP. In mice, the enzyme is found on early B-lineage cells and certain stromal cells of the bone marrow and thymus. This ectopeptidase is also expressed by capillary endothelial cells, placenta, and epithelial cells of the intestine and proximal renal tubules. Here we have used a mouse E-AP cDNA to identify the human counterpart in a kidney library. Sequence comparison of the human and mouse cDNAs reveals approximately 80% homology at both nucleotide and predicted amino acid levels. The nucleotide sequence of human E-AP predicts a type II integral membrane protein of 957 amino acids with an 18-amino-acid aminoterminal intracellular domain, and a 22-amino-acid transmembrane domain. The large extracellular carboxyterminal domain contains the zinc-binding motif typical of zinc-dependent metallohydrolases. When the human E-AP cDNA was placed downstream of the SR alpha promoter in an expression vector and transfected into COS-7 cells, the transfected cells exhibited cell surface E-AP activity. A 4.1-kb transcript could be detected in a variety of human tissues, including heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas. However, in representative lymphoid leukemias, E-AP transcripts were restricted to pre-B leukemia and were not found in T- and B-cell leukemias. The cDNA cloning and successful expression of human E-AP will allow more precise analysis of its physiological role(s). PMID- 8244383 TI - Physical delineation of the minimal chromosomal segment encompassing the murine host resistance locus Bcg. AB - The host resistance locus Bcg determines resistance of mice to infection with intracellular pathogens such as certain species of Mycobacteria, Salmonella typhimurium, and Leishmania donovani. Bcg maps on the proximal portion of mouse chromosome 1, very tightly linked to villin (Vil), with the gene order and intergene distances Tp-1-(1 cM)-D1Mcg105-(0.1 cM)-lambda Mm1C165/Vil/Bcg-(0.2 cM) lambda Mm1C136-(0.3 cM)-Des-(0.1 cM)-Inha. In an effort to clone genomic sequences overlapping Bcg, we have used pulsed-field gel electrophoresis (PFGE) and fluorescence in situ hybridization to construct a physical map of the 3.9-Mb segment of proximal mouse chromosome 1, near Bcg. In situ hybridization to metaphase mouse chromosomes indicates that the mapped region is within band C5. Physical mapping of the Tp-1-Vil and lambda Mm1C136-Inha intervals was carried out by PFGE analysis, whereas the Vil-Des interval was estimated by in situ hybridization to interphase nuclei. Results of these combined analyses indicate the following locus order and maximal interlocus distances: Tp-1-(1000 kb) D1Mcg105-(160 kb)-lambda Mm1C165-(180 kb)-Vil-(800 kb)-lambda Mm1C136-(290 kb) Des-(130 kb)-Inha. Detailed restriction mapping of this region identifies numerous putative CpG islands, suggesting that several transcription units are present in the vicinity of Bcg. PMID- 8244384 TI - Three human glutamate dehydrogenase genes (GLUD1, GLUDP2, and GLUDP3) are located on chromosome 10q, but are not closely physically linked. AB - Yeast artificial chromosomes (YACs) of 340 and 370 kb that contain the functional human glutamate dehydrogenase gene (GLUD1) and the pseudogene GLUDP2, respectively, were isolated. These genes were not physically linked to each other nor to any other sequences homologous to the exons of GLUD1. No additional GLUD sequences were found within at least 70 kb of the 5' and 175 kb of the 3' end of GLUD1 or 150 kb of either end of GLUDP2. By in situ hybridization, GLUD1 was located at 10q23.3, GLUDP2 at 10q11.2, and another pseudogene of the GLUD gene family, GLUDP3, at 10q22.1. DNA fragments of these three genes showed cross hybridization to the loci assigned to the other two genes, but not to any other chromosomal locus. Thus, these three genes are located at distinct positions on chromosome 10q. PMID- 8244385 TI - Characterization of two chromosome 12 cosmid libraries and development of STSs from cosmids mapped by FISH. AB - We have constructed and characterized two related human chromosome 12-specific cosmid libraries. DNA from flow-sorted chromosomes from a somatic cell hybrid was cloned into a cosmid vector. Approximately 61% of the cosmids in the nearly 26,200 member arrayed libraries (LL12NC01 and LL12NC02) contain human DNA inserts, and 31% of the cosmids derived from human DNA contain CA repeats. One hundred and fifty-two cosmids isolated from the libraries have been mapped by fluorescence in situ hybridization (FISH). Cosmids containing human DNA inserts were localized by FISH exclusively to chromosome 12, confirming the chromosomal specificity of the libraries. The cosmids have been localized to all parts of this chromosome, although some regions are more highly represented than others. Partial sequence information was obtained from 44 mapped cosmids, and oligonucleotide primer pairs were synthesized that define unique sequence tagged sites (STSs). These mapped cosmids, and unique STSs derived from them, provide a set of useful clones and primer pairs for screening YAC libraries and developing contigs centered on regions of interest within chromosome 12. In addition, 120 of the mapped cosmids contain CA repeats, and thus they also provide a useful resource for defining highly polymorphic simple tandem repeat elements that serve as genetic markers for linkage analysis and disease gene localization. PMID- 8244386 TI - Physical and linkage mapping of the human and murine genes for the alpha 1 chain of type IX collagen (COL9A1). AB - Type IX collagen, a member of the FACIT family of extracellular matrix proteins, is a heterotrimer composed of three genetically distinct alpha chains. The cDNAs for the human and mouse alpha 1 (IX) chains have been cloned. In this paper we confirm the mapping of the human COL9A1 gene to chromosome 6q12-q13 by fluorescence in situ hybridization utilizing two genomic clones which also contain short tandem repeat polymorphisms. We also report the characterization of these repeats and their incorporation into the chromosome 6 linkage map. The COL9A1 locus shows no recombination with the marker D6Z1 (Z = 27.61 at theta = O) and identifies the most likely locus order of KRAS1P-[D6Z1-COL9A1]-D6S30. In addition, using an interspecific backcross panel, we have mapped murine Col9a1 to mouse chromosome 1. Together with other comparative mapping results, these data suggest that the pericentric region of human chromosome 6 is homologous to the most proximal segment of mouse chromosome 1. These data may facilitate linkage studies with COL9A1 (or Col9a1) as a candidate gene for hereditary chondrodysplasias and osteoarthritis. PMID- 8244387 TI - A sequence-tagged site map of human chromosome 11. AB - We report the construction of 370 sequence-tagged sites (STSs) that are detectable by PCR amplification under sets of standardized conditions and that have been regionally mapped to human chromosome 11. DNA sequences were determined by sequencing directly from cosmid templates using primers complementary to T3 and T7 promoters present in the cloning vector. Oligonucleotide PCR primers were predicted by computer and tested using a battery of genomic DNAs. Cosmids were regionally localized on chromosome 11 by using fluorescence in situ hybridization or by analyzing a somatic cell hybrid panel. Additional STSs corresponding to known genes and markers on chromosome 11 were also produced under the same series of standardized conditions. The resulting STSs provide uniform coverage of chromosome 11 with an average spacing of 340 kb. The DNA sequence determined for use in STS production corresponds to about 0.1% (116 kb) of chromosome 11 and has been analyzed for the presence of repetitive sequences, similarities to known genes and motifs, and possible exons. Computer analysis of this sequence has identified and therefore mapped at least eight new genes on chromosome 11. PMID- 8244388 TI - TSPY-related sequences represent a microheterogeneous gene family organized as constitutive elements in DYZ5 tandem repeat units on the human Y chromosome. AB - TSPY (testis-specific protein, Y-encoded) is encoded by members of a Y-chromosome specific sequence family. We show here that TSPY elements are part of the DYZ5 repeat unit. We have established a cosmid library of Y-chromosomal DNA derived from the hybrid cell line 3E7 and isolated eight cosmids representing 15 TSPY elements. Interindividual variability with respect to TSPY copy number was observed. One cosmid clone was investigated in detail by subcloning and sequencing. Sequence microheterogeneity was identified among both transcribed and nontranscribed TSPY elements. Transcript variability is not restricted to single basepair exchanges. We present data for the existence of at least three differently sized TSPY transcripts, caused by different patterns of splicing. PMID- 8244389 TI - A bovine homologue of the human TSPY gene. AB - We have isolated a male-specific bovine genomic homologue of the human TSPY gene (testis-specific protein, Y-encoded). Two specific transcripts can be detected in bovine testicular RNA. The cloned sequence represents one copy of a clustered array of some 50-200 related sequences on the Y chromosome resembling the arrangement present in the human genome. The clone isolated from the bovine genome has two regions that are highly conserved between homo and bos. One stretch of DNA is homologous in 45 of 51 bp (88%), and the other stretch shares 71 of 100 bp (71%). PMID- 8244390 TI - Evidence that the SRY protein is encoded by a single exon on the human Y chromosome. AB - To facilitate studies of the SRY gene, a 4741-bp portion of the sex-determining region of the human Y chromosome was sequenced and characterized. Two RNAs were found to hybridize to this genomic segment, one transcript deriving from SRY and the second cross-hybridizing to a pseudogene located 2.5 kb 5' of the SRY open reading frame (ORF). Analysis of the SRY transcript using 3' and 5' rapid amplification and cloning of ends suggested that the entire SRY protein is encoded by a single exon. A 700-bp CpG island is located immediately 5' of the pseudogene (and 2 kb 5' of the SRY ORF). Within this CpG island lies the sequence CGCCCCGC, a potential binding site for the EGR-1/WT1 family of transcription factors, some of which appear to function in gonadal development. PMID- 8244391 TI - A leucine-to-proline substitution causes a defective alpha 1-antichymotrypsin allele associated with familial obstructive lung disease. AB - Using denaturing gradient gel electrophoresis and direct sequencing of amplified genomic DNA, we have identified two defective mutants of the human alpha 1 antichymotrypsin (ACT) gene associated with chronic obstructive pulmonary disease (COPD). A leucine 55-to-proline substitution causing a defective ACT allele (Bochum-1) was observed in a family with COPD in three subsequent generations. Another mutation, proline 229-to-alanine (Bonn-1), was associated with ACT serum deficiency in four patients with a positive family history. These mutations were not detected among 100 healthy control subjects, suggesting a possible pathogenetic role of ACT gene defects in a subset of patients with COPD. PMID- 8244392 TI - A panel of sequence tagged sites for chromosome band 11q23. AB - A panel of sequence tagged sites (STSs) representing 30 markers previously assigned to human chromosome band 11q23 has been assembled. Eleven STSs represent cloned genes, and the remainder are from anonymous DNA segments. The STSs have been used in PCR experiments to localize their cognate sequences further with respect to five translocation breakpoints that define three intervals in 11q23. Two of these translocation breakpoints have been mapped more precisely by the STS assignments. The STS panel will form a useful starting point for the generation of a genomic contig of band 11q23. PMID- 8244393 TI - Assignment of the human Na+/glucose cotransporter gene SGLT1 to chromosome 22q13.1. AB - The Na+/glucose cotransporter gene SGLT1 encodes the primary carrier protein responsible for the uptake of the dietary sugars glucose and galactose from the intestinal lumen. SGLT1 transport activity is currently exploited in oral rehydration therapy. The 75-kDa glycoprotein is localized in the brush border of the intestinal epithelium and is predicted to comprise 12 membrane spans. In two patients with the autosomal recessive disease glucose/galactose malabsorption, the underlying cause was found to be a missense mutation in SGLT1, and the Asp28- >Asn change was demonstrated in vitro to eliminate SGLT1 transport activity. The SGLT1 gene was previously shown to reside on the distal q arm of chromosome 22 (11.2-->qter). We have used a cosmid probe for fluorescence in situ hybridization, which refines the localization to 22q13.1, and provide an example of the utility of the SGLT1 probe as a diagnostic for genetic diseases associated with translocations of chromosome 22. PMID- 8244394 TI - The human prohibitin (PHB) gene family and its somatic mutations in human tumors. AB - Five cosmid clones, isolated by procedures to screen genomic libraries for homologous variants of the human prohibitin gene (PHB), were analyzed to determine their genomic structures. Four of these (PHBP1-4) were found to be processed pseudogenes, each located on a different chromosome from their counterparts on chromosome 17q21. The DNA sequence of one clone (PHBP1, on chromosome 6q25) shared a 91.3% identity at the nucleotide level with the cDNA of functional prohibitin. A large number of human tumors of the breast, ovary, liver, and lung were examined for somatic mutations in the PHB gene. Although mutations were observed in a few sporadic breast cancers, none were identified in any of the other cancers. PMID- 8244395 TI - The prostatic acid phosphatase (ACPP) gene is localized to human chromosome 3q21 q23. AB - Human prostatic acid phosphatase (ACPP) has been used as a diagnostic marker for prostate cancer. It is synthesized under androgen regulation and secreted by the epithelial cells of the prostate gland. We have confirmed the previous assignment of the ACPP gene to chromosome 3 by probing a panel of 25 human-Chinese hamster somatic cell hybrids, and we have further localized the ACPP gene to chromosome 3q21-q23 by fluorescence in situ hybridization. PMID- 8244396 TI - Highly polymorphic tetramer repeat (GATA)n on human chromosome 11p15.3. AB - A tetrameric repeat sequence (GATA)n was identified by random DNA sequencing of chromosome 11-specific cosmid clones and located at 11p15.3 by fluorescence in situ hybridization. Oligonucleotide primers flanking the repetitive unit were used to amplify the DNA using the polymerase chain reaction (PCR) and this repetitive element was shown to be highly polymorphic and inherited in typical Mendelian fashion. Analysis of amplification products containing the repetitive element from 45 unrelated Caucasian individuals and five families showed at least five alleles at this locus, ranging from 227 to 249 bases in length. This polymorphism may serve as a useful PCR-detectable genetic marker for 11p15.3, a landmark for disease gene isolation, and a locus for identity testing. PMID- 8244397 TI - Sequence of the human iduronate 2-sulfatase (IDS) gene. AB - Deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS; EC 3.1.6.13) results in the storage of the glycosaminoglycans heparan sulfate and dermatan sulfate, which leads to the lysosomal storage disorder mucopolysaccharidosis type II. Three overlapping genomic clones derived from an X-chromosome-specific library containing the entire IDS gene were isolated and the sequences of the intron boundaries and the 5' promoter region were determined. The IDS gene is split into nine exons spanning approximately 24 kb. The potential promoter for IDS lacks a TATA box but contains GC box consensus sequences, consistent with its role as a housekeeping gene. A polypyrimidine-like repeat is found in intron 1. PMID- 8244398 TI - Isolation and mapping of 45 NotI linking clones to chromosome 22. AB - Fifty-nine NotI linking clones have been isolated from a flow-sorted chromosome 22 cosmid library and mapped using fluorescence in situ hybridization and/or a panel of somatic cell hybrids. Fourteen clones map to the short arm of chromosome 22, 31 to the long arm, and 9 to other chromosomes; 5 clones could not be unambiguously mapped. To identify potentially informative genetic markers, the chromosome 22 clones were screened for poly(CA) sequences; 24 positively hybridizing clones, 10 on the long arm and 14 on the short arm, were identified. These clones will be useful for constructing a long-range restriction map of chromosome 22 and may facilitate the cloning of chromosome 22 genes. PMID- 8244399 TI - Mapping of the glycoprotein 330 (Gp330) gene to mouse chromosome 2. AB - Glycoprotein 330 (Gp330) is a member of the low-density lipoprotein receptor gene family that is expressed in the kidney. We have mapped the Gp330 gene to mouse chromosome 2, 4.5 cM proximal to Acra, in an interspecific backcross of (C57BL/6J x Mus spretus) F1 x C57BL/6J. PMID- 8244400 TI - Localization of the gene for the ciliary neurotrophic factor receptor (CNTFR) to human chromosome 9. AB - Ciliary neurotrophic factor (CNTF) has recently been found to be important for the survival of motor neurons and has shown activity in animal models of amyotrophic lateral sclerosis (ALS). CNTF therefore holds promise as a treatment for ALS, and it and its receptor (CNTFR) are candidates for a gene involved in familial ALS. The CNTFR gene was mapped to chromosome 9 by PCR on a panel of human/CHO somatic cell hybrids and localized to 9p13 by PCR on a panel of radiation hybrids. PMID- 8244401 TI - Human somatostatin receptor genes: localization of SSTR5 to human chromosome 20p11.2. AB - The gene encoding the somatostatin receptor subtype designated as SSTR5 was mapped to human chromosome 20p11.2 by using fluorescence in situ hybridization to metaphase chromosomes. Fluorescence in situ hybridization using a probe for SSTR5 in combination with probes for neuroendocrine convertase-2 (NEC2), thrombomodulin (THBD), and brain glycogen phosphorylase (PYGB) established a physical order for these loci of 20pter-NEC2-SSTR5-THBD-PYGB-cen. PMID- 8244402 TI - Localization of the Na+/glucose cotransporter gene SGLT2 to human chromosome 16 close to the centromere. AB - The chromosomal location of the gene SGLT2, which is presumed to encode a low affinity Na+/glucose cotransporter, has been determined using a panel of rodent human somatic cell hybrids. Southern blot analysis of genomic DNA from 16 different hybrids shows that SGLT2 is located on chromosome 16. Analysis of three additional hybrids that selectively retain all or part of human chromosome 16 demonstrates that SGLT2 is located close to the centromere in band p11.2 of the chromosome. PMID- 8244403 TI - Molecular cloning and chromosomal localization to 17q21 of the human WNT3 gene. AB - In mouse mammary tumors, the Wnt-3 gene can be activated by proviral insertion. Here we report on the isolation of a human homolog, WNT3. A genomic clone was isolated by use of mouse Wnt-3 sequences as a probe, after which cDNA containing most of the protein-encoding domain of the human gene was obtained by PCR. Comparison between the deduced mouse and human WNT-3 protein sequences showed four changes in 333 amino acids. WNT3 is located on chromosome 17q21. The gene was not found to be amplified or rearranged in a collection of human breast tumors. PMID- 8244404 TI - Localization of the human Sp1 transcription factor gene to 12q13 by fluorescence in situ hybridization. PMID- 8244405 TI - Genetic mapping of the high-affinity nerve growth factor receptor gene, Ntrk1, to mouse chromosome 3. PMID- 8244406 TI - Rapid identification of VNTR alleles of the human thyroid peroxidase gene by PCR: a study in a population sample from south Italy. PMID- 8244407 TI - A subset of 1200 hexamers is sufficient to sequence over 95% of cDNAs by hexamer string primer walking. PMID- 8244408 TI - Aggregation bias and the use of regression in evaluating models of human performance. AB - Regression analyses are increasingly being used to provide confirmatory evidence for models of human performance. The amount of information made available to judge these models is reduced because clearly established standards in the techniques of performing and reporting regression analyses are lacking. This paper addresses two primary problems in regression analysis: aggregation of data and the aggregation of variables into composite models. We provide examples of the misuse of regression techniques and recommend ways in which the amount of information made available to evaluate the model being tested can be maximized in analysis and reporting. PMID- 8244409 TI - Experimental evaluation of a model of mental workload. AB - This study was designed to test predictions from a model of mental workload. The model predicts that mental workload grows as perceived distance from a task goal increases and the effective time for action decreases. Decreases in workload can be achieved by actions that bring the task goal into the region of acceptable time/distance constraints for successful resolution. We reported an experiment that tested these assertions using the Timepools performance task. Timepools generates a spatial representation of a shrinking temporal target to assess the effects of path length (i.e., the number of sequential targets to be acquired) and shrink rate (i.e., elapsed time during which the circle is halved in area) on reaction time (RT), movement time (MT), error rate, and the subjective perception of workload. Data from the experiment indicated systematic effects for task related factors across performance and workload measures. Path length and shrink rate had differential effects on both RT and MT, which were also reflected in the components of the individual workload scales. The results support a general form of the workload model which may help researchers and practitioners in the difficult task of workload prediction. PMID- 8244410 TI - Spatial and temporal characteristics of rapid cursor-positioning movements with electromechanical mice in human-computer interaction. AB - This research examines how people make movements with pointing devices during human-computer interaction. It specifically concerns the perceptual-motor processes that mediate the speed and accuracy of cursor positioning with electromechanical mice. In three experiments we investigated the spatial and temporal characteristics of positioning movements made with a mouse, analyzing subjects' speed and accuracy as a function of the types of targets that the movements had to reach. Experiment 1 required rapid and accurate horizontal movements to targets that were vertical ribbons located at various distances from the mouse's starting location. The targets for Experiments 2 and 3, respectively, were vertical lines having various heights and rectangular boxes having various heights and widths. Constraints on movement distance along the primary (that is, horizontal) line of motion had the greatest effects on total positioning times. However, constraints on movement distance along a secondary (vertical) line of motion also affected total positioning times significantly. These effects may be localized in different phases of movements (e.g., movement execution and verification). The duration of movement execution (i.e., physical motion) depends primarily on the target distance, whereas the duration of movement verification (i.e., check for endpoint accuracy) depends primarily on target height and width. A useful account of movement execution is provided by stochastic optimized submovement models, which have significant implications for designing mice and menu-driven displays. PMID- 8244411 TI - Effects of stereoscopic and rotational displays in a three-dimensional path tracing task. AB - A series of three experiments investigated the effectiveness of stereoscopic and rotational display techniques for the purpose of establishing human factors guidelines for the design of three-dimensional (3D) displays. In the described experiments, depth perception was evaluated by examining accuracy in a 3D path tracing task, with stimulus displays resembling the structure of cerebral angiograms. The first experiment allowed subjects to control rotation in dynamic displays. The results indicated that performance improved using either technique relative to viewing two-dimensional (2D) displays. However, rotational displays were superior to stereoscopic displays, and performance was best when both techniques were combined. The second experiment compared subject-controlled rotation with observation of continuously rotating displays at different rates of rotation. Performance declined at faster rotation rates; however, there were no advantages of subject-controlled rotation. In the third experiment, performance in rotational displays was no better than that in stereoscopic displays enhanced with multiple static viewing angles. However, performance was always best when both 3D techniques were jointly implemented. The results are discussed in terms of the visual information available using either 3D display technique and are related to the weighted additive model of depth perception. PMID- 8244412 TI - Spatial ability and land navigation under degraded visual conditions. AB - Land navigation tasks require the use of visual cues. When these cues are degraded by the loss of resolution, navigators suffer varying degrees of performance decrements. We tested the hypothesis that these decrements are less severe for people of high spatial ability than they are for people of low spatial ability. We tested 108 noncommissioned officers on a task that required them to determine if two woodland photographs taken from different directions (N, NE, E ... NW) were of the same location; spatial ability was assessed using the Cognitive Laterality Battery. Spatial ability was related to the ability to do this task. Furthermore, there was a significant interaction between spatial ability and visual resolution on recognition performance. This article discusses the implications of these results for teleoperations and land navigation. PMID- 8244413 TI - Curvilinear approach to an intersection and visual detection of a collision. AB - Visual motion perception plays a fundamental role in vehicle control. Recent studies have shown that the pattern of optical flow resulting from the observer's self-motion through a stable environment is used by the observer to accurately control his or her movements. However, little is known about the perception of another vehicle during self-motion--for instance, when a car driver approaches an intersection with traffic. In a series of experiments using visual simulations of car driving, we show that observers are able to detect the presence of a moving object during self-motion. However, the perception of the other car's trajectory appears to be strongly dependent on environmental factors, such as the presence of a road sign near the intersection or the shape of the road. These results suggest that local and global visual factors determine the perception of a car's trajectory during self-motion. PMID- 8244414 TI - Pistol grip power tool handle and trigger size effects on grip exertions and operator preference. AB - Finger and palmar forces were measured during actual pneumatic nutrunner operation using a strain gauge dynamometer. Eighteen student subjects were assigned to one of three categories based on hand length. Two triggers and four handle spans were presented randomly. Handle span affected maximal and submaximal grip force. As span increased from 4 cm to 7 cm, average peak finger force increased 24%, peak palmar force increased 22%, and average finger and palmar tool-holding forces increased 20%. When an extended trigger was used, average peak finger force decreased 9%, peak palmar force decreased 8%, finger tool holding force decreased 65%, and palmar tool-holding force decreased 48%. Hand size affected grip strength (MVC), grip force, and exertion level (force/MVC). Holding exertion level was maximum for large-handed subjects using a 4-cm handle and for small-handed subjects using a 7-cm handle. Subjective handle span preference increased as hand size increased. A similar experiment was performed using 11 factory workers. PMID- 8244415 TI - Inhibition of matrix metalloproteinase 9 activation by a specific monoclonal antibody. AB - Two members of the matrix metalloproteinase (MMP)1 family of enzymes are expressed at elevated levels in highly aggressive human tumor cells and have been implicated in the catalytic functions of extracellular proteolysis. The zymogen forms of these enzymes are designated proMMP-2 and proMMP-9, also known as 72kDa and 92kDa type IV collagenases/gelatinases, respectively. The MMP family of enzymes can be activated in vitro by a number of compounds including the organomercurial 4-aminophenylmercuric acetate (APMA). The natural or in vivo activators of MMP-2 and MMP-9 are at present unknown. A partially purified preparation of MMP-9 was used to immunize mice for the isolation of monoclonal antibodies (mAbs). Three IgG1 mAbs were identified by immunoreactivity with purified MMP-9 and are designated 6-6B, 7-11C, and 8-3H. These mAbs react specifically with MMP-9 by ELISA and Western blot. Additionally, these mAbs react with N-glycanase treated 92kDa protein. These mAbs were tested for their ability to inhibit enzyme activation in a radio-labeled gelatin assay. The 6-6B mAb inhibited the activation of MMP-9, but had no effect on MMP-2. These mAbs are highly specific to human MMP-9 and the 6-6B mAb will be extremely useful for examining the autolytic and catalytic activity of MMP-9 in normal and abnormal biological processes. PMID- 8244416 TI - Construction and expression of two mouse-human chimeric antibodies with high specificity and affinity for carcinoembryonic antigen. AB - We have previously reported that a group of monoclonal antibodies (MAbs) to carcinoembryonic antigen (CEA), designated Group F MAbs, are able to discriminate CEA in tumor tissues from the CEA-related normal antigens and that CEA assay systems utilizing at least one Group F MAb show the improved cancer diagnosis. In this study, we cloned the genes coding for two Group F MAbs (F11-35 and F11-39) and deduced the amino acid sequences of the variable regions for their heavy and light chains. The variable region for the heavy chain of F11-35 contained a possible N-glycosylation site (Asn/Asp/Thr) at amino acid positions 89-91. Then, we constructed two mouse-human chimeric antibodies by using the F11-35 and F11-39 variable region genes of heavy and light chains (VH and V kappa) and human heavy and light chain constant region genes (gamma 1 and kappa) derived from a human plasma cell leukemia line (ARH77). The chimeric gene constructs were sequentially co-transfected into murine non-Ig-producing myeloma (P3-U1) or hybridoma (Sp2/0) cells by electroporation. The resulting chimeric heavy chain of F11-35 showed a slightly but significantly higher molecular weight than that of F11-39, but the molecular weights of their unglycosylated peptides synthesized in the presence of tunicamycin were similar, indicating the glycosylation at the possible N glycosylation site in the variable region of the Ch F11-35 heavy chain. Both chimeric antibodies exhibited the same specificity and affinity for CEA as those of the parental murine hybridoma antibodies, respectively. Ascites production of Sp2/0 transfectomas is sufficiently high (600-900 micrograms/ml) for initial clinical studies with the chimeric antibodies. PMID- 8244417 TI - Isolation and characterization of monoclonal antibodies to a recombinant human topoisomerase II polypeptide. AB - We have produced two murine monoclonal antibodies (SWT3D1 and SWR1C2) to a recombinant polypeptide corresponding to the carboxyl-terminal one-third (amino acid 854-amino acid 1447) of human topoisomerase II alpha. Each antibody is able to recognize intact human topoisomerase II using immunoblotting and enzyme-linked immunosorbent assay (ELISA) techniques. Data is presented demonstrating that the antibodies bind specifically to topoisomerase II alpha but do not interact with topoisomerase II beta. The monoclonal antibodies do not recognize murine or calf thymus topoisomerase II indicating that each may bind exclusively to the human enzyme. The topoisomerase II binding sites for each monoclonal antibody have been compared in a competition ELISA. The SWT3D1 antibody had no significant effect on the binding efficiency of biotinylated SWR1C2 antibody. Although SWR1C2 was capable of inhibiting the binding of biotinylated SWT3D1, this only occurred at concentrations approximately 1000-fold higher than those required of SWT3D1 to block binding of itself. These results suggest that SWT3D1 and SWR1C2 do not recognize identical epitopes on topoisomerase II. PMID- 8244418 TI - Production and characterization of monoclonal antibodies to the carboxyl-terminal heptapeptide of endothelin-1. AB - Two cell lines, RR5.ET-1 and RR1.ET-1, that produce monoclonal antibodies specific for the carboxyl-terminal heptapeptide of endothelin-1 (ET-1) have been cloned and stabilized. An RIA was developed to facilitate the evaluation and characterization of these monoclonal antibodies. The affinity constant of each MAb for ET-1, as determined by Scatchard analysis, was 5.74 x 10(8) M-1 for RR5.ET-1 and 4.15 x 10(7) M-1 for RR1.ET-1. The antibodies reacted specifically with the carboxyl-terminus (ET15-21) and did not cross-react with the amino terminal amino acids (ET1-16). As expected, the antibodies cross-reacted with endothelin-2 (ET-2) and endothelin-3 (ET-3), and to a lesser extent, with the closely related sarafotoxins. Both MAbs retained about 55% reactivity with the ET 1 terminal sequence of Asp-Ile-Ile-Trp (ET18-21) but had no reactivity with the ET sequence His-Leu-Asp-Ile-Ile-Trp-Val-Asn (ET16-23) nor with Big ET-1 (ET1-39). These data strongly suggest that the terminal four amino acids of ET-1 are included in the MAb binding site. More importantly, the terminal Trp21 must be free, not linked to Val22 to retain reactivity with either of the MAbs. PMID- 8244419 TI - Novel immunization protocol and ELISA screening methods used to obtain and characterize monoclonal antibodies specific for human light chain variable-region subgroups. AB - We have developed a novel immunization protocol for the production of a panel of high-affinity murine monoclonal antibodies (MoAbs) that are specific for each of the major human kappa and lambda light chain variable-region (VL) subgroups. Mice were injected with heat-precipitated human Bence Jones proteins or VL-related fragments emulsified in monophosphoryl lipid A (MPL) and trehalose dimycolate (TDM) at two- to four-week intervals over a seven-month period. A unique direct capturing enzyme-linked immunosorbent assay (ELISA) employing biotinylated monoclonal light chains was designed to select optimally immunized animals for hybridoma preparation and to screen culture supernatants for high-affinity anti VL MoAbs. These methods have led to the generation of MoAbs that by ELISA react specifically with each of the four V kappa subgroups--V kappa I, V kappa II, V kappa III, and V kappa IV or five V lambda subgroups--V lambda I, V lambda II/V, V lambda III, V lambda IV, and V lambda VI. These reagents have been used successfully to establish, on the basis of VL subgroup, the monoclonal nature of serum or urinary immunoglobulins as well as those found in the cytoplasm or on the cell surface of monoclonal plasma cell or B-lymphocyte populations, respectively. The availability of anti-VL subgroup-specific MoAbs will facilitate the immunodiagnosis and study of patients with multiple myeloma, AL amyloidosis, and related B-cell proliferative disorders. PMID- 8244420 TI - Cloning of cDNAs encoding the variable domains of antibody KC4G3 and construction of a chimeric antibody. AB - We have cloned and sequenced cDNAs encoding the variable regions of the light (VL) and heavy (VH) chains of monoclonal antibody KC4G3. VL belongs to group II and resulted from a V kappa-J kappa 2 recombination. VH belongs to group IIId and arose from a V-D9-JH3 recombination. The VL and VH frameworks are respectively 84% and 83% identical to the corresponding VL and VH human consensus frameworks. The deduced amino acid sequence of VL contains an asparagine-linked glycosylation site in framework 3 (N74 I75 S76). We have determined that a large fraction of the light chains are indeed glycolysated. We constructed an IgG1, kappa human/mouse chimeric antibody (by inserting the murine KC4G3 Fv-encoding cDNAs into plasmids encoding a human IgG1, kappa Fc domain) and expressed it in SP2/0 Ag14 mouse myeloma cells. This chimeric monoclonal antibody is designated ChiKC4. We have determined that the murine monoclonal antibody KC4G3 binds the human breast mucin with an affinity constant of 1.1 x 10(9) M-1. ChiKC4 binds the same antigen with an affinity constant of 1.2 x 10(9) M-1. ChiKC4 binds the carcinoma tissue sections by the ABC immunoperoxidase method in an identical manner as does KC4G3. PMID- 8244421 TI - A test for teratological effects of power frequency magnetic fields on chick embryos. AB - An analysis of 13 studies of the teratological effects of pulsed magnetic fields on chick embryos from ten independent laboratories permits no clear conclusions. Comparatively little has been done to follow up on the reports by Juutilainen and coworkers on the effects of extremely low-frequency, sinusoidal magnetic fields on the malformation rate in chick embryos. Our attempt to follow up on their results using similar but not identical exposures of 10 microT, 50 Hz magnetic fields produced negative results. PMID- 8244422 TI - Multiple multipole method for simulating EM problems involving biological bodies. AB - The three-dimensional implementation of the multiple multipole (MMP) method, based on the generalized multipole technique (GMT), is presented. Its performance in simulating electromagnetic problems involving biological bodies is analyzed. In particular, the step-by-step simulation technique and the built-in procedures to validate the solution on numerical basis are discussed and demonstrated in two examples. A comparison is made with other numerical techniques often applied in this field. The advantages of the MMP method are shown to be in its validation capability, in its efficiency for smoothly shaped bodies and in the achievable accuracy, in particular near boundaries. The method is especially suited to handle high-gradient fields in the vicinity of biological bodies. On the other hand, finite difference (FD) techniques are superior for scatterers with complicated angular shapes or inhomogeneous bodies, for which MMP shows rather strong practical limitations. However, in most cases the inhomogeneities of biological bodies modify the field distribution only locally beyond the uncertainties of models. In these cases, inhomogeneities can be stimulated efficiently and with high accuracy by MMP applying the block iterative technique. Other methods are not general enough to compete with FD or MMP in solving EM problems involving biological tissues. PMID- 8244423 TI - Boundary element analysis of the directional sensitivity of the concentric EMG electrode. AB - Assessment of the motor unit architecture based on concentric electrode motor unit potentials requires a thorough understanding of the recording characteristics of the concentric EMG electrode. Previous simulation studies have attempted to include the effect of EMG electrodes on the recorded waveforms by uniformly averaging the tissue potential at the coordinates of one- or two dimensional electrode models. By employing the boundary element method, this paper improves earlier models of the concentric EMG electrode by including an accurate geometric representation of the electrode, as well as the mutual electrical influence between the electrode surfaces. A three-dimensional sensitivity function is defined from which information about the preferential direction of sensitivity, blind spots, phase changes, rate of attenuation, and range of pick-up radius can be derived. The study focuses on the intrinsic features linked to the geometry of the electrode. The results show that the cannula perturbs the potential distribution significantly. The core and the cannula electrodes measure potentials of the same order of magnitude in all of the pick-up range, except adjacent to the central wire, where the latter dominates the sensitivity function. The preferential directions of sensitivity are determined by the amount of geometric offset between the individual sensitivity functions of the core and the cannula. The sensitivity function also reveals a complicated pattern of phase changes in the pick-up range. Potentials from fibers located behind the tip or along the cannula are recorded with reversed polarity compared to those located in front of the tip. Rotation of the electrode about its axis was found to alter the duration, the peak-to-peak amplitude, and the rise time of waveforms recorded from a moving dipole. PMID- 8244424 TI - Excitation of dorsal root fibers in spinal cord stimulation: a theoretical study. AB - In epidural spinal cord stimulation it is likely that not only dorsal column fibers are activated, but that dorsal root fibers will be involved as well. In this investigation a volume conductor model of the spinal cord was used and dorsal root fibers were modeled by an electrical network including fiber excitation. The effects of varying some geometrical fiber characteristics, as well as the influence of the dorsal cerebrospinal fluid layer and the electrode configuration on the threshold stimulus for their excitation, were assessed. The threshold values were compared with those of dorsal column fibers. The results of this modeling study predict that, besides the well known influence of fiber diameter, the curvature of the dorsal root fibers and the angle between these fibers and the spinal cord axis were of major influence on their threshold values. Because of these effects, threshold stimuli of dorsal root fibers were relatively low as compared to dorsal column fibers. Excitation of the dorsal root fibers occurred near the entry point of the fibers. PMID- 8244425 TI - Selective control of muscle activation with a multipolar nerve cuff electrode. AB - Acute experiments were performed on adult cats to study selective activation of medial gastrocnemius, soleus, tibialis anterior, and extensor digitorum longus with a cuff electrode. A spiral nerve cuff containing twelve "dot" electrodes was implanted around the sciatic nerve and evoked muscle twitch forces were recorded in six experiments. Spatially isolated "dot" electrodes in four geometries: monopolar, longitudinal tripolar, tripolar with four common anodes, and two parallel tripoles, were combined with transverse field steering current(s) from an anode(s) located 180 degrees around from the cathode(s) to activate different regions of the nerve trunk. To quantify the degree of selectivity, a selectivity index was defined as the ratio of the force in one muscle to the force in all four muscles in response to a particular stimulus. The selectivity index was used to construct recruitment curves for a muscle with the optimal degree of selectivity. Physiological responses were correlated with the anatomical structure of the sciatic nerve by identifying the nerve fascicles innervating the four muscles, and by determining the relative positions of the electrodes and the nerve fascicles. The results indicated that the use of transverse field steering current improved selectivity. We also found that tripoles with individual dot anodes were more selective than tripoles with four common dot anodes. Stimulation with two parallel tripoles was effective in activating selectively fascicles that could not be activated selectively with only a single tripole. The multipolar cuff proved an effective method to control selectively and progressively the force in muscles innervated by fascicles that were well defined at the level of the cuff. PMID- 8244426 TI - Axon termination conditions for electrical stimulation. AB - The cable model for electrical stimulation near the terminal of a passive fiber is derived for excitation by an arbitrary, time-varying, applied extracellular field. Unless the termination impedance is comparable to that of mammalian node of Ranvier, the end-conditions require the longitudinal intracellular current at the fiber terminal to be negligibly small. This requirement substantially alters the membrane potential profile from that obtained with a fiber of infinite length. Stimulation near the end of a fiber may result in lower thresholds and may reverse the anodal/cathodal threshold ratio obtained with stimulation in the mid-portion of the fiber. Chronaxie for stimulation near the terminal may be much smaller than at a distance from the terminal and the strength-duration curve may be nonmonotonic. These differences may have significant implications for any application of electrical stimulation where fiber terminations may play a role in the excitatory process. PMID- 8244427 TI - A musculotendon model of the fatigue profiles of paralyzed quadriceps muscle under FES. AB - In a previous work, we studied the mechanical and the metabolic profiles of fatigue of paralyzed quadriceps muscle under activation by FES. The metabolic state of the muscle during stimulation of paraplegic patients was monitored, simultaneously with the decaying force, by using 31P nuclear magnetic resonance spectroscopy. In the present work, a musculotendon model was developed to enable prediction of the force output during continuous electrical stimulation. The model consisted of five elements, including the tendon, the parallel elastic, contractile and damper muscle elements, as well as the muscle mass. The mechanism of the contractile element was based upon the length-tension and the velocity tension curves, the activation trajectory, and the experimentally obtained relationship between force and intracellular pH. In the equations obtained, three sets of parameters were used: 1) general muscle parameters, associated with the length-tension curves of tendon, fascia, and muscle and the velocity-tension curve of the contractile element; 2) specific anthropometric parameters of the muscle; and 3) fatigue parameters which were obtained from our previously recorded experimental data. The model was formulated to allow prediction of the quadriceps muscle force under dynamic activation and at various levels of stimulation. The model solution was for isometric contraction in supermaximal stimulation, and it provided the force decaying profiles, which were compared to those obtained experimentally. The parameters yielding the best fit between the model and the experimental results were indicated. Particularly, two muscle nonspecific parameters, namely, the muscle stress parameter and the parameter representing the ratio between the muscle's slack length and its length in vivo at various knee angles, were determined using the model. The muscle stress parameter was found to be between 60 and 64 N/cm2, and the length ratio was 0.952, 0.935, 0.920, and 0.901 for the 0, 30, 60, and 90 degrees knee angle, respectively. Finally, a sensitivity analysis was conducted of the model to perturbations of these two estimated parameters, revealing that the model was sensitive to these parameters. PMID- 8244428 TI - Automated tuning of a closed-loop hand grasp neuroprosthesis. AB - An automated tuning algorithm was developed to reduce the time and skill required to tune a closed-loop hand grasp neuroprosthesis. The time reduction results from simultaneous tuning of four gain parameters controlling the dynamic response of the system, and from automation of the calculation and decision processes. The new tuning method is therefore an automated parallel tuning method, replacing a manual sequential method in which only one parameter at a time was tuned. RMS error between the step input and the grasp output is minimized, with absence of oscillation as a constraint. The difference between the system's RMS ramp tracking errors for the two tuning methods was less than 1% of the ramp size regardless of the initial values of the parameters, implying that the tuning methods were equivalent. However, the parallel tuning method was faster and required fewer trials than the sequential method. The capability of the closed loop system to regulate grasp output in the presence of disturbances was compared with the capability without feedback. Patients were instructed to either grasp an object at a certain force level or to match a certain grasp opening. They would then lock their command at a fixed value, and either remain immobile to test time dependence or pronate and supinate their forearm to test postural disturbances. With closed-loop control, the grasp output was better regulated in the presence of disturbances, with an average output variance 60% lower than without feedback control. PMID- 8244429 TI - Generation of two-dimensional spatial and temporal properties through spatiotemporal convergence between one-dimensional neurons. AB - Primary otolith afferents are characterized by diverse temporal and spatial response properties. The temporal properties of these neurons vary from tonic to phasic response characteristics during stimulation with linear acceleration. This presentation examines the response properties of target neurons that arise from spatiotemporal convergence (STC) between purely tonic and phasic-tonic afferents. The transfer function of the phasic-tonic afferent is described by either fractional leaky differentiator or integrator terms. Target neurons would generally exhibit two-dimensional spatial sensitivity and are characterized by two perpendicular response vectors. It is shown that target neurons have different temporal properties during stimulation along different spatial directions. Specifically, they could exhibit tonic temporal response dynamics during stimulation along one response vector and phasic dynamics during stimulation along the second response vector. The phasic dynamics along one response vector are described by a complete ideal differentiator for frequencies below the corner frequency of the leaky operator terms. PMID- 8244430 TI - A moment-based three-dimensional edge operator. AB - This paper presents a three-dimensional edge operator aimed at the detection of anatomical structures in medical imaging. It uses the spatial moments of gray level surface, and operates in three dimensions with any window size. It allows us to estimate the location and the contrast surface, as well as the surface orientation. The computation of the discrete version is reported. Bias and errors due to the spatial sampling and noise are analyzed both at a theoretical and experimental level. The moment-based operator is compared with other well-known edge operators on simple shaped primitives for which the analytical solution is known. The 3D rendering of real data is then provided by merging the operator in a ray-tracing framework. PMID- 8244431 TI - Mechanics of the occlusive arm cuff and its application as a volume sensor. AB - Although a common medical instrument, the mechanical function of an occlusive arm cuff has not been fully described in an engineering sense. The occlusive arm cuff is examined here using a mathematical mechanics model and experimental measurements. Cuff stretch was modeled by a nonlinear pressure-volume function. Air compression was represented by Boyle's law. An apparatus was developed to measure pressure due to the air volume pumped into the cuff for fixed arm volume. Data were obtained for two different cuff designs, and reveal a nonlinear cuff pressure-volume relationship that could be represented accurately by the mathematical model. Calibration constants are provided for the two types of occlusive cuff. Thus, the cuff pressure was found to consist of a balance between that produced by stretch of the elastic cuff bladder and that of the compression of the air contained within the bladder. The use of the gas law alone was found to be inadequate to represent the cuff mechanics. When applying the cuff to measure change in arm volume, such as during plethysmography or oscillometry, it cannot be assumed that the cuff sensitivity is constant. More precisely, it was found that the occlusive cuff is a transducer with a volume sensitivity that increases with cuff pressure and volume until it becomes nearly constant at high levels of cuff pressure (150 mmHg). A hypothetical case of a linear elastic artery with constant pulse pressure was used as input to the cuff model to illustrate the change in cuff pressure oscillations that occurs while cuff pressure is released.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244432 TI - A system for the exposure of small laboratory animals to a 25-mT 60-Hz alternating or traveling magnetic field. AB - This paper describes a versatile exposure system which can be used to investigate the effects of a 60-Hz alternating magnetic field or traveling magnetic field on small laboratory animals. The system was designed and built as part of an extensive experimental effort to find out what effects, if any, these fields would have on the various biological systems of animals. Special attention was given to the design of the electric circuit to simplify the supply requirements and to keep the overall current to a minimum. Magnetic field having a flux density up to 30 mT (300 G) can be generated, and a traveling magnetic field is obtained by reconnecting the coils to a three-phase supply. The total magnetic field due to the currents in the three coils used in the exposure system is analyzed by considering the x, y, and z components of the flux density due to each coil, and then using the method of superposition to obtain the flux density B at any point in the region of the three coils. A computer program was written to evaluate the magnetic field in the region where the animals are located. The computed values are also compared with those obtained from actual measurements. The difference between the measured and the computed values of the flux density is attributed to the idealized representation of the multiturn, multilayer coils by equivalent single-turn coils. PMID- 8244433 TI - High permittivity patch radiator for single and multi-element hyperthermia applicators. AB - This paper describes a compact, low-profile patch radiator which is the base element for efficient, small-size applicators suitable for superficial hyperthermia. The design criteria and the technological processes involved are presented. The electromagnetic characteristics of the patch element are outlined, and possible application of the radiator are discussed. PMID- 8244434 TI - Comments on "Radio-frequency coils in implantable devices: misalignment analysis and design procedure". PMID- 8244435 TI - Decline in edentulism means more practice activity. PMID- 8244436 TI - Final rules--continuing education for dentists and dental hygienists. PMID- 8244437 TI - Fluoride overfeeds in public water supplies. AB - A description of the fluoride overfeed in Hooper Bay, Alaska, that resulted in the first reported death due to fluoride toxicity caused by drinking water from a community water system. PMID- 8244438 TI - Selecting an effective telephone answering strategy. PMID- 8244439 TI - Overview of appliance therapy in sleep apnea and snoring. PMID- 8244440 TI - HIV-1 gp41 binding to human peripheral blood mononuclear cells occurs preferentially to B Lymphocytes and monocytes. AB - Based on our findings that HIV-1 gp41 independently of CD4 can bind to the human helper T lymphoid cell line H9, B cell line Raji and monocyte cell line U937, we characterized putative binding of HIV-1 gp41 to human peripheral blood lymphocytes (PBLs) and monocytes. Using flow cytometry (FACS), we demonstrated that the recombinant soluble HIV-1 gp41 (sgp41; Env amino acid 539-684) can bind to the normal human peripheral blood mononuclear cells (PBMCs), preferentially to B lymphocytes and monocytes independently of gp120-binding sites on CD4 molecules. This binding is dose-dependent. The HIV-1 sgp41 binds to blood B lymphocytes and monocytes more strongly than to T lymphocytes. By two-color flow cytometric analysis, we identified that sgp41 can bind 10% of CD4+ T lymphocytes, 11.9% of CD8+ T lymphocytes, 47% of CD19+ B lymphocytes and 44.2% of CD14+ monocytes. PMID- 8244441 TI - Analysis of T cell receptor V beta regions expressed by rheumatoid synovial T lymphocytes. AB - The T cell receptor (TCR) V beta gene segment repertoire of T lymphocytes derived from peripheral blood of two healthy individuals and synovial tissue, synovial fluid and peripheral blood of three rheumatoid arthritis (RA) patients was analyzed. A sensitive assay based on the amplification of cDNA by the polymerase chain reaction (PCR) was used to analyze the levels of expression of 20 TCR V beta gene segment families. The relative expression of V beta gene segments in lymphocytes derived from peripheral blood, synovial tissue and synovial fluid was conserved over 155 days in one patient. V beta 9 transcripts were undetectable in the cells of this individual. In the two other patients the frequency of V beta 2 transcripts in synovial T cells of affected joints was significantly higher than in their peripheral blood lymphocytes. Dominance of distinct rearrangements among the V beta 2 transcripts from the synovial cells of these patients support the idea that the synovial T cell response is driven by antigen. PMID- 8244442 TI - Analyses of thymic abnormalities in autoimmune-prone (NZW x BXSB) F1 mice. AB - Thymic abnormalities of autoimmune-prone (NZW x BXSB)F1 mice are investigated. After the onset of autoimmune diseases, severe thymic atrophy is observed. The atrophied thymus shows lower CD4+ CD8+ T cell and higher CD4+CD8- or CD4-CD8+ T cell counts than aged-matched normal strains or other autoimmune-prone (NZB x NZW)F1(B/WF1) mice; even at the age of 40 weeks, B/WF1 mice have a large number of double-positive cells and a small number of single-positive cells. Thymocytes in the atrophic thymus of (NZW x BXSB)F1 mice respond better than normal mice to T cell-mitogens (PHA and ConA). In addition, the single-positive T cells are J 11d-negative. These findings indicate that the atrophic thymus includes mature T cells and a large number of plasma cells and B cells; it therefore responds well to lipopolysaccharides. Experiments in reciprocal transplantation of the thymus and bone marrow between (NZW x BXSB)F1 and normal mice show that the thymic abnormalities are due to defects of the hemopoietic stem cells (HSCs) rather than either intrinsic abnormalities in the thymus or extrinsic abnormalities such as anti-thymus antibodies. It should be noted that the atrophic thymus recovers after transplantation of normal bone marrow cells; the atrophic thymus still has the capacity to induce the differentiation of normal T cells (including double positive T cells) if normal HSCs are introduced. PMID- 8244443 TI - The effect of adult thymectomy on immune response to infection with Listeria monocytogenes in mice. AB - The role of adult thymus in in vivo immune response to infection with Listeria monocytogenes was examined using euthymic mice and adult-thymectomized (ATx) mice which had been thymectomized 2 weeks before. Numbers of T cells in peritoneal cavities and spleens were increased at 2 weeks after inoculation of L. monocytogenes, whereas such increases of T cells were several times higher in euthymic mice than in ATx mice. In flow-cytometric analysis of peritoneal exudate cells, a significant increase of CD3+CD4-CD8- cells bearing gamma/delta type T cell receptor was noted in euthymic mouse after infection compared to ATx mouse. Neither assay of antigen-specific T cell proliferation nor analysis of cell cycle exhibited any superiority of T cells obtained from euthymic mice to those obtained from ATx mice. These findings suggest that the enlargement of T cell population in euthymic mice is attributed largely to T cells which emigrate from the thymus after inoculation of L. monocytogenes. Moreover, in vivo protective immune response against secondarily challenged L. monocytogenes was achieved more efficiently in euthymic mice than in ATx mice, as shown by the clearance of the bacteria from organs and the survival rate of infected mice. Our results indicate the importance of adult thymus-dependent increase of T cells in eliminating the facultative intracellular bacteria such as L. monocytogenes. PMID- 8244444 TI - Immune restoration in children after partial splenectomy. AB - Splenectomy (SE) is recognized to be a therapeutical approach in treating children with severe autoimmune diseases (chronic idiopathic thrombocytopenia; hemolytic anemia) or hypersplenism because of portal hypertension. Nevertheless, removal of a main immune organ results in elevated infection risk for these patients. Partial splenectomy (PSE) was developed as a therapeutical compromise to retain immunologically active spleen tissue. Here, we document the analysis of immune parameters obtained from children after both partial and total splenectomy, which have been followed up for a period of more than 6 years: (i) Lymphocytes from both groups of patients failed to produce IgG in response to pokeweed mitogen in vitro. This was observed in 11/20 splenectomized patients even 10 years after operation, whereas in PSE patients a restoration of this parameter after 1-2 years was seen. (ii) In patients after PSE, but not in splenectomized persons, an elevated number of HLA-class II positive cells had been detected suggesting a different situation of immune regulation following this operation. However, in parallel with an improvement of B cell in vitro activity this parameter was found to achieve normal values. Our findings indicate that partial splenectomy may be a therapeutical alternative, if the therapeutic goal can be achieved by this procedure. PMID- 8244445 TI - Effects of IL-4 depletion on the antibody response to Pseudomonas aeruginosa lipopolysaccharide in mice. AB - These studies were done to examine the role of interleukin-4 (IL-4) in the generation of isotype specific antibody responses of mice to Pseudomonas aeruginosa lipopolysaccharide (PALPS) by neutralization of IL-4 in vivo using anti-IL-4 antibody (11B11). We found that the administration of anti-IL-4 antibody (11B11) 24 h before immunization with PALPS resulted in a decreased PALPS-specific antibody response for all isotypes examined (IgM, IgG1, IgG2a, IgG2b, IgG3). By contrast, we observed that the non-antigen-specific (polyclonal) IgM response of mice following treatment with 11B11 antibody and PALPS was increased while the polyclonal responses for the other isotypes were unaffected. When mice were given recombinant IL-10 at the time of immunization with PALPS there was a decrease in the PALPS-specific antibody response but an increase in the polyclonal IgM, IgG2a, IgG2b, IgG3 response whereas the polyclonal IgG1 response was decreased by a five-fold margin. The results of these studies suggest that both the antigen-specific and the polyclonal response can be influenced in a different manner by IL-4 or by IL-10. PMID- 8244446 TI - Characterization of new monoclonal antibodies identifying avian T lymphocyte antigens. AB - Six monoclonal antibodies (mAb) were produced to identify and characterize surface antigens of chicken T cells. Determination of their reactivity with different lymphatic cells using immunofluorescence analysis demonstrates that mAb KH8, NA6, PD4 and TH8 stained 32-43% blood lymphocytes, 72-77% thymocytes and 19 27% spleen cells, mAb OC5 approximately 99% thymocytes and 55% blood and spleen lymphocytes each, and mAb OC2 36% blood lymphocytes, 79% thymocytes and 62% spleen cells. The KH8, NA6, PD4 and TH8 antibodies immunoprecipitated from lysates of surface-labeled chicken thymocytes a polypeptide of M(r) 60,000 under non-reducing conditions and the OC5 antibody a glycoprotein of M(r) 68,000 under reducing conditions. MAb OC2 precipitated a single polypeptide of M(r) 40,000 under both conditions. The mAb KH8, NA6, PD4, TH8 and OC2 inhibited ConA-induced proliferative responses of blood T cells in vitro. However, sepharose-bound or soluble OC5 antibody was able to increase DNA synthesis significantly. These results indicate that (a) the mAb KH8, NA6, PD4 and TH8 identify the avian homologue of the mammalian CD4 molecule, (b) the mAb OC2 detects the avian CD2 antigen, and (c) the mAb OC5 recognizes the putative avian CD5 homologue. PMID- 8244447 TI - Tumor-induced macrophage tumor necrosis factor-alpha production suppresses autoreactive T cell proliferation. AB - T cells can react to self-cells bearing the syngenic major histocompatibility complex class II molecule Ia. Decreased autoreactive T cell responses are associated with cancer. Tumor growth causes syngeneic macrophages (M phi) to suppress autoreactive T cell proliferation by decreasing M phi Ia expression and increasing M phi production of the suppressor molecule prostaglandin E2 (PGE2). Because M phi produce tumor necrosis factor-alpha (TNF-alpha) during cancer, and TNF-alpha stimulates M phi PGE2 synthesis, we determined if TNF-alpha mediates tumor-induced suppression of autoreactive T cell proliferation stimulated by syngeneic M phi. We showed that tumor growth increases TNF-alpha production because tumor-bearing host (TBH) M phi synthesized more TNF-alpha than normal host (NH) M phi when cultured with lipopolysaccharide. Exogenous TNF-alpha increased NH CD4+ autoreactive T cell proliferation stimulated by syngeneic NH M phi but not by TBH M phi. When endogenous TNF-alpha activity was neutralized by anti-TNF-alpha antibody addition, T cell proliferation decreased when stimulated by NH M phi but increased when stimulated by TBH M phi. Kinetic studies showed that TNF-alpha affected M phi-stimulated T cell proliferation during the first few hours (4h) of the 96 h culture time. Indomethacin-treatment allowed TNF-alpha to increase T cell proliferation stimulated by TBH M phi. A PGE2-specific enzyme linked immunosorbent assay showed that TBH M phi T cell cultures contained significantly more PGE2 than those containing NH M phi, and that exogenous TNF alpha increased PGE2 production in TBH M phi cultures more than in NH M phi cultures. Short-term (4h) pretreatment of M phi with TNF-alpha increased T cell proliferation stimulated by NH, but not TBH, M phi. However, long-term (16 h) TNF alpha pretreatment reversed TBH M phi-mediated suppression, suggesting that early suppressor molecular production inhibits synthesis or activity of TNF-alpha induced stimulatory monokines. Although TNF-alpha is known to increase T cell proliferation, these results show that the tumor-induced increase in M phi TNF alpha synthesis suppress autoreactive T cell proliferation, which is mediated by PGE2 production. PMID- 8244448 TI - IL-1, TNF-alpha and IL-2 production by peritoneal and spleen cells from Schistosoma mansoni infected mice and its potentiation by preimmunization with schistosomal antigens and immunostimulants. AB - In the present study we tested the effect of immunization with schistosome derived antigens such as frozen-thawed schistosomula in combination with either BCG, liposomes or liposomal muramyl tripeptide-phosphatidyl ethanolamine (MTP PE), on the resistance of mice to infection, and on the function of their macrophages and lymphocytes. Immunization with either F-T schistosomula + BCG or F-T schistosomula + MTP-PE and subsequent infection, resulted in a 2-3-fold increase in adherent peritoneal macrophage-mediated schistosomulicidal activity (SCA). Peritoneal and spleen macrophages from immunostimulant treated and/or immunized animals showed a significant increase in LPS triggered TNF-alpha production, as compared to non-treated controls. The highest increase in TNF alpha production was achieved after immunization with either F-T schistosomula + BCG or F-T schistosomula + MTP-PE. LPS triggered IL-1 production was elevated in spleen and peritoneal macrophages from F-T schistosomula + BCG treated mice, and also in spleen macrophages treated with F-T schistosomula + MTP-PE. Only immunization with F-T schistosomula + BCG increased ConA-induced spleen lymphocyte proliferation and IL-2 production. Immunization of mice with F-T schistosomula + BCG also induced protection against parasite infection, while F-T schistosomula + MTP-PE failed to do so. Potentiation of antischistosomal resistance seems to require both macrophage and lymphocyte activation which was achieved only when BCG served as an immunostimulant. PMID- 8244449 TI - Retinoic acid induces changes in c-fgr proto-oncogene mRNA levels in Burkitt's lymphoma cells. AB - The c-fgr proto-oncogene is expressed in Burkitt's lymphoma (BL) cell and cell lines derived from them. When Epstein-Barr virus (EBV)-negative BL cell lines that contain low levels of c-fgr mRNA are infected with EBV, transcription of the c-fgr gene is further induced. In this paper we show that treatment of EBV negative and EBV-positive BL cell lines with all-trans retinoic acid also stimulates an increase in c-fgr mRNA levels, varying between 2- and 13-fold depending on the cell line. An increase is detectable 12 to 48 h after treatment, depending on the cell line, suggesting that the c-fgr gene is not regulated directly by retinoic acid but responds to other retinoic acid-induced changes in the cell. We also show that treatment of BL cell lines with all-trans retinoic acid either results in a dose-dependent decrease in growth rate, or has no effect on growth, depending on the cell line. It has previously been suggested that the c-fgr gene product might have a role in regulating the growth of BL cells, since treatment of the EBV-positive BL cell line Daudi with alpha-interferon results in a decrease in c-fgr mRNA levels followed by a decrease in growth rate. Our data indicate that there is no general correlation between c-fgr mRNA levels and growth rate in BL cells and so argue against a role for the c-fgr gene product in growth regulation in these cells. PMID- 8244450 TI - A role for interleukin-1 alpha in immunologically mediated intestinal pathology. AB - Interleukin-1 (IL-1) is an important mediator of inflammation and has been implicated in several forms of immunopathology. Here we have investigated whether IL-1 plays a role in the enteropathy which occurs during a graft-versus-host reaction (GVHR) in mice. Non-irradiated (CBA x BALB/c) F1 mice with GVHR had increased production of IL-1 and treatment with rabbit anti-IL-1 alpha antibodies abolished the crypt hyperplasia and significantly reduced the parallel increase in crypt length which occurs in the jejunum. Antibody treatment had no effect on the accompanying increase in intraepithelial lymphocyte (IEL) counts or on the splenomegaly. Recombinant IL-1 itself produced villus atrophy, crypt hyperplasia and increased IEL counts in normal mice and stimulated the proliferation of an intestinal epithelial cell line in vitro. We propose that IL-1 plays an effector role in immunologically mediated enteropathy, either via direct effects on epithelial cells or secondary to an action on other, stromal cells in the mucosa. PMID- 8244451 TI - Intestinal protection against Strongyloides ratti and mastocytosis induced by administration of interleukin-3 in mice. AB - Information about interleukin-3 (IL-3) effects in vivo is limited compared with the in vitro effects. We found that a repetitive injection of a low dose of recombinant IL-3 induced protection against intestinal worms of Strongyloides ratti in C57BL/6 mice. When mice were injected i.p. with different doses of recombinant IL-3 twice a day from day -5 to day -1 and infected orally with larvae recovered from the head of infected rats on day 0, worm recovery from the small intestine was markedly reduced by a total of 10(4) U IL-3 or more on day 2 post-infection. The number of intestinal mucosal mast cells (MMC) was increased by the protective dose of IL-3. The IL-3 treatment, however, was ineffective in protecting mice against tissue migrating larvae, as assessed by recovery from the head. The protective effect of IL-3 on intestinal worms was observed within 6 hr post oral infection, suggesting little concern with antigen-specific immune responses. The effective dose of IL-3 treatment increased the number of MMC progenitors five times in the spleen and the mesenteric lymph nodes. An MMC specific protease, MMCP-1, was secreted 200 times more than in controls in the intestinal lumen by the IL-3 treatment. The IL-3 treatment induced no protection or mastocytosis in mast cell-deficient W/Wv mice. These results suggest that the IL-3-induced intestinal protection against S. ratti is mediated by MMC. PMID- 8244452 TI - Chloroquine inhibits macrophage tumour necrosis factor-alpha mRNA transcription. AB - Although chloroquine administration in vivo following haemorrhage in mice decreases tumour necrosis factor-alpha (TNF-alpha) release by macrophage (M phi), the mechanism remains unknown. To study this, peritoneal M phi (pM phi) from unmanipulated, sham-operated and post-haemorrhage mice were isolated, treated with 0.13 mg/ml chloroquine for 2 hr, and then stimulated with lipopolysaccharide (LPS) for 48 hr. Pretreatment of pM phi from various groups of mice with chloroquine resulted in 75-90% inhibition of TNF-alpha release, determined by bioassay. Total RNA was isolated from pM phi and murine M phi-derived cell lines (P388D1 and RAW 264.7), stimulated with LPS for 0.5 or 1 hr, respectively, and Northern blot analysis for TNF-alpha mRNA performed. Chloroquine inhibited TNF alpha mRNA expression without interfering with mRNA stability, suggesting that this agent reduces M phi TNF-alpha release by disrupting TNF-alpha gene transcription. PMID- 8244453 TI - Chloroquine-induced inhibition of the production of TNF, but not of IL-6, is affected by disruption of iron metabolism. AB - There is now considerable evidence that cerebral malaria may be related to the over-production of tumour necrosis factor (TNF). Nevertheless, our knowledge is very poor concerning the biological events which lead up to this TNF over production. Furthermore, interleukin-6 (IL-6) is produced in large amounts during malaria infection and seems to have inhibitory action on TNF production. Anti malarial drugs were investigated for their ability to interfere with TNF and IL-6 secretion by human non-immune macrophages stimulated by lipopolysaccharides (LPS) or Plasmodium falciparum culture supernatant. Macrophages were pretreated with chloroquine, quinine, proguanil, mefloquine or halofantrine before stimulation. TNF and IL-6 production were suppressed in a dose-dependent manner when macrophages were treated with chloroquine, but not with other anti-malarial drugs. Considering that chloroquine probably acts via lysosomotropic mechanisms, and that iron metabolism may interfere with the non-specific immune response, we focused our attention on these biochemical events in order to investigate the mechanisms by which chloroquine inhibits cytokine production. Our results demonstrated that chloroquine-induced inhibition of TNF and IL-6 production is not mediated through a lysosomotropic mechanism, and that chloroquine probably acts on TNF secretion by disrupting iron homeostasis. Inhibition of IL-6 production seems not to be mediated through these pathways. These observations suggest that chloroquine may help to prevent cerebral malaria whatever the drug sensitivity of the parasite strain, and may provide new tools for an anti-disease therapy regardless of the emergence of parasite multi-drug resistance. PMID- 8244454 TI - Characterization of autoreactive helper T cells in a murine model of autoimmune haemolytic disease. AB - Repeated immunization of mice with rat red blood cells (RRBC) results in the production of both erythrocyte autoantibodies and anti-RRBC antibodies. The manner in which erythrocyte self-tolerance is broken has been little studied. It has been assumed that help for autoreactive B cells is provided by Th cells specific for the foreign RRBC. We show here that autoreactive Th cells can be recovered from RRBC-immunized mice. The Th cells proliferate in vitro whether stimulated by self or rat erythrocytes. Analysis of the specificity of the proliferating cells revealed extensive cross-reactivity for the two types of erythrocytes. It is therefore surprising that, in initial cultures, a slower response is evident when mouse erythrocytes are used as the antigen. From cytotoxic depletion of T-cell subsets, the phenotype of the proliferating cells was identified as Thy-1.2+, Lyt-1.1+, Lyt-2.1-, L3T4+. During in vitro stimulation of the T cells, growth factors characteristic of Th cells are secreted. Finally, we demonstrate that the responding T cells are able to help primary in vitro antibody responses to self and rat erythrocytes. We conclude that autoreactive Th cells are likely to be involved in the experimental induction of autoimmune haemolytic disease in mice. PMID- 8244455 TI - Pro-tumour necrosis factor cleavage enzyme in macrophage membrane/particulate. AB - Tumour necrosis factor (TNF) is synthesized initially as a membrane-bound precursor which is then cleaved to yield soluble, mature protein. The 26,000 MW TNF precursor isolated from the lysate of activated RAW 264.7 (mouse macrophage) cells by immunoprecipitation was used to identify pro-TNF cleavage enzyme in the same cells. A significant amount of mature protein was formed in samples containing Nonidet P-40 (NP-40)-lysed cells, whereas sonicated cells showed negligible activity. Most of the cleavage activity in macrophages was localized in the membrane/particulate fraction and remained largely insoluble after sonication or treatment with 2 mM EDTA/1 M NaCl, indicating that the enzyme is associated with the membrane/particulate fraction. The crude cleavage activity in membrane/particulate was partially inhibited by a spectrum of serine, cysteine and aspartate proteinase inhibitors, whereas secretion of TNF from activated macrophages was inhibited exclusively by serine and serine/cysteine proteinase inhibitors. This result suggested that, among heterogenous pro-TNF cleavage activities, the enzyme responsible for the processing of TNF is a serine proteinase. Pro-TNF cleavage activity was present in non-stimulated macrophages and decreased significantly 8 hr after lipopolysaccharide (LPS) stimulation, suggesting that it is negatively regulated after an initial burst of TNF synthesis. PMID- 8244456 TI - Thrombin generates monocyte chemotactic activity from complement factor H. AB - We have recently found that the complement factor H (H) was the precursor of the major macrophage chemotactic factor in the delayed-type hypersensitivity (DTH) reaction site in the skin and was converted to the factor by an unidentified trypsin-like protease in plasma. Thrombin and plasmin are also present in the site, and we, therefore, examined the possibility that these proteases converted H to be monocyte chemotactic. Intact H caused no monocyte migration, although it was able to do so after incubation with thrombin, but not with plasmin. The activity was chemotactic rather than chemokinetic and was absorbed by an anti-H IgG-conjugated column. The generation of monocyte chemotactic activity from H was dependent on incubation time with thrombin and also the protease activity of thrombin, and the activity was seen at concentrations of H lower than 10(-8) M. The inhibitory activity of H for C3b-Bb was not affected by incubation with thrombin or plasmin. Incubation of H with thrombin, but not with plasmin, generated a hydrophobic molecule, in a time-dependent manner, which had monocyte chemotactic activity. These results show that H becomes a monocyte chemotactic factor due to cleavage by thrombin, which converts H to a more hydrophobic molecule and also suggest that thrombin-treated H induces monocyte migration in the DTH reaction site. PMID- 8244457 TI - Differential regulation of cytokine production by nitric oxide. AB - Nitric oxide (NO) has recently been identified as a potent and pleiotropic intracellular mediator produced by and acting on many cells of the body. Although considerable attention has been devoted to the regulation of NO by inflammatory cytokines, and also to the role of NO as an important effector molecule in immune function, there is very little information on the role of this mediator in modulating T-cell-dependent cytokine production. In this study we show that physiological levels of NO (either produced by activated macrophages or by the addition of exogenous NO donors) can selectively down-regulate interleukin-3 (IL 3) production by spleen cells from contact-sensitized mice, while leaving IL-2 activity unaffected. Thus NO may have an important role as an immunomodulatory as well as effector molecule in the immune system. PMID- 8244458 TI - Pentoxifylline: a potent inhibitor of IL-2 and IFN-gamma biosynthesis and BCG induced cytotoxicity. AB - In the present study we investigated the influence of pentoxifylline (POF) on bacillus Calmette-Guerin (BCG)- and phytohaemagglutinin (PHA)-induced DNA synthesis and cytokine release, and BCG-induced cytotoxicity of human peripheral blood mononuclear cells (PBMC). DNA synthesis of PBMC stimulated with either BCG or PHA was inhibited by POF. We also demonstrated that the addition of POF led to a POF dose-dependent decrease of the release of the cytokines interleukin (IL)-2, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha). The release of IL-6 remained unaffected. With respect to the inhibition of BCG induced IL-2 and IFN-gamma release POF is active at the transcriptional (mRNA) level, as found by polymerase chain reaction (PCR). However, PHA-induced mRNA expression of these lymphokines is not affected by POF. Thus, the existence of a post-transcriptional regulation of PHA-induced cytokine release by POF can be assumed. The observed inhibition of cytokine release is correlated with a potent inhibitory effect of POF on BCG-induced cytotoxicity against bladder tumour cell lines. This effect is reversible. PMID- 8244459 TI - Amino acid sequence of the region of beta 2-glycoprotein 1 (gp1) which mediates binding of autoantibodies to the cardiolipin-gp1 complex in humans. AB - Anticardiolipin antibodies (ACA) in sera from patients with autoimmune and infectious diseases were tested for binding to beta 2-glycoprotein 1 (gp1) in order to determine whether human gp1 acts as a cofactor for the binding of ACA to cardiolipin (CL) or as an antigen recognized by ACA. While none of the ACA positive sera tested recognized gp1 by itself, gp1 was necessary for the binding of ACA to CL in sera from four patients with autoimmune diseases. In three of the four sera the presence of lupus anticoagulant (LA) was detected by prolonged partial thromboplastin time (PTT). Examinations using the bovine equivalent of human gp1 contained in fetal calf serum (FCS) and adult bovine serum (ABS) showed that the human protein can be replaced by the bovine equivalent in the enzyme linked immunosorbent assay (ELISA). Using affinity-purified antibodies directed against the CL-gp1 complex it was shown that the binding of these antibodies is dependent on the concentration of the bovine gp1 equivalent contained in the formed complex. Similar results found with the human gp1 confirmed this assertion. In order to find out which region of gp1 might mediate the binding between ACA and cardiolipin, we examined to what extent selected oligopeptide sequences of gp1 can substitute for the protein. Peptide P2 (representing the amino acids at positions 268-278 of the gp1 molecule) and gp1 showed about the same binding capacity. Histidine in this peptide seems to be essential for the binding to CL as we found decreased binding with peptides modified in this position. Conclusions from this work show that gp1 does not act as a relevant antigen for ACA, but occupies an essential function in the complex formed with cardiolipin for a certain group of ACA. PMID- 8244460 TI - The role of various compartments in the chicken spleen during an antigen-specific humoral response. AB - In the chicken spleen, certain aspects of the complex of ellipsoid, peri ellipsoid lymphocyte sheath (PELS), and ring of macrophages are morphologically and functionally similar to the marginal zone in the mammalian spleen. In this study, the role of this marginal zone-like complex was examined during the humoral immune response against a thymus-dependent antigen. The antigen peroxidase-conjugated rabbit anti-peroxidase (PAP) was injected intravenously in chickens and subsequently visualized by its enzymatic activity. Anti-PAP antibody containing cells (ACC) were detected by incubation with PAP and subsequent peroxidase staining. Shortly after injection, PAP antigen was located in the ellipsoids. Within 24 hr most PAP had relocated to the B lymphocytes of the PELS. After this, most PAP disappeared, although minute amounts of PAP were detected in peri-arteriolar T-cell areas (PALS). At 48 hr, the PALS contained the first anti PAP ACC; later ACC were also located in the red pulp. During the immune response, the isotype of the ACC switched from IgM to IgG. From 72 hr, PAP antigen and anti PAP antibodies were trapped on follicular dendritic cells in immature and mature germinal centres. These results demonstrate that humoral immune responses are induced by the interaction of the PALS and the complex of ellipsoid, PELS and macrophages. These results, along with earlier ones, suggest that this complex is analogous to the marginal zone in the mammalian spleen. PMID- 8244461 TI - Two functionally different follicular dendritic cells in secondary lymphoid follicles of mouse spleen, as revealed by CR1/2 and FcR gamma II-mediated immune complex trapping. AB - The capacity to trap immune complexes (IC) was examined using purified peroxidase antiperoxidase IgG on frozen sections of mouse spleen. In the absence of serum, binding of IC was confined to macrophages in the red pulp. However, when IC were applied in the presence of fresh mouse serum as a source of complement, trapping occurred on follicular dendritic cells (FDC) in the primary follicles. Trapping was specifically inhibited with monoclonal antibodies against complement receptor type 1 (CR1) (8C12) and CR 1/2 (7G6), but not with anti-CR3 (Mac-1). The binding mechanism of IC in the secondary follicles of animals immunized with sheep red blood cells differed from that observed in unimmunized mice. Here, trapping occurred both in the absence and presence of fresh mouse serum. In the absence of fresh mouse serum, trapping on FDC was mediated by FcR gamma II, as demonstrated by blocking with monoclonal antibody 2.4G2. FcR gamma II-mediated trapping was only observed on a relatively small proportion of FDC that was located in the light zone of the secondary follicles, while complement receptor-mediated IC trapping was observed in more expanding areas, including dark zone and corona. These results show that both complement receptor as well as Fc receptor can function in the binding of IC to FDC. They also provide evidence for the existence of functionally different FDC populations in the secondary lymphoid follicles of mouse spleen. PMID- 8244462 TI - Surface expression of immunoglobulin isotypes on primary human B cells: no evidence for glycosyl-phosphatidylinositol linkage. AB - Surface expression of membrane(m) IgM molecules requires the association of two disulphide-linked transmembrane (TM) glycoproteins, encoded by the B-cell specific genes mb-1 and B29. We have shown that mIgM, mIgD and mIgG are associated with structurally related heterodimers on primary human B cells. Transfection studies in murine plasmacytoma cells, however, have demonstrated mb 1-independent expression of both mIgD and mIgG. The recent finding that mIgD is expressed on these cells through glycosyl-phosphatidylinositol (GPI) linkage may be interesting in view of the function of mIgD on primary B cells. We therefore investigated whether GPI linkage serves as an additional mechanism for expression of mIgD and the other mIg isotypes on primary human B cells. However, we were unable to demonstrate the release of mIg molecules upon treatment with phosphatidylinositol-specific phospholipase C (PI-PLC) in either immunofluorescence analysis or Ig heavy (H) chain-specific enzyme-linked immunosorbent assay (ELISA). We conclude that primary human B cells, which constitutively express the mb-1 and B29 genes, do not express the mIg isotypes in a GPI-linked manner. PMID- 8244463 TI - Non-oscillatory free Ca2+ response of single B cells and WEHI-231 cells after cross-linking of antigen receptors with anti-immunoglobulin. AB - Cross-linking of surface-immunoglobulin (sIg) has been associated with IP3 production and a rise in cytoplasmic-free calcium ([Ca2+]i) in studies of populations of normal and transformed B cells. We have examined the kinetics of the induced cytoplasmic calcium rises in single, Fura-2-loaded cells, during stimulation with a variety of agonists. Our data indicate that the responses of B cells to some stimuli, such as elevated cyclic AMP, consist of repetitive calcium transients, but these Ca2+ oscillations do not occur after sIg ligation. Rather, sIg cross-linking leads to a rapid rise in cytoplasmic calcium which is followed by a sustained elevation. Most of this response seems to result from an inflow of extracellular calcium rather than from internal stores. PMID- 8244464 TI - Expression and functional properties of mouse B7/BB1 using a fusion protein between mouse CTLA4 and human gamma 1. AB - We report the construction and expression of a fusion protein made from the extracellular portion of the mouse CTLA-4 gene and the constant region of human IgG1. This fusion protein behaves like an antibody to mouse B7/BB1, binding to activated B cells and purified dendritic cells. In addition, we found it to bind to activated T cells. The fusion protein interfered with the ability of antigen pulsed antigen-presenting cells to induce proliferation of T-cell clones, although the degree of inhibition varied. These findings are discussed in the light of the physiological activation of T cells in secondary lymphoid organs. PMID- 8244465 TI - Suppression of murine IgM, IgG, IgA and IgE antibody responses by alveolar macrophages. AB - Freshly recovered pulmonary alveolar macrophages (AM) and the AM-derived established cell line, MH-S, have previously been shown to be highly suppressive of in vitro IgM anti-sheep erythrocyte (SRBC) responses. Supernatants obtained from cultures of AM incubated with antigen-stimulated lymphocytes or from the MH S cell line alone have also been shown to be suppressive when added to the in vitro antibody-forming system. In order to determine if AM and MH-S cells, owing to their mucosal location, could differentially regulate antibody responses including immunoglobulin isotypes other than IgM, an in vitro system for the detection of cells producing IgG, IgA and IgE anti-2,4 dinitrophenol (DNP) antibody was developed. These studies demonstrate that AM, MH-S cells, and MH-S culture supernatants suppress the in vitro generation of IgM, IgG, IgA and IgE anti-DNP spot-forming cells (SFC). No apparent differential regulation of any of the four murine IgG anti-DNP antibody subclasses was observed. Time-course experiments suggested that optimal AM- and MH-S-mediated suppression occurred 18 hr after culture initiation. Both AM and MH-S cells suppressed IgM and IgG anti DNP antibody responses in a dose-related manner, suggesting that MH-S is a good model for the study of AM-mediated immunoregulation. PMID- 8244466 TI - Biological markers of macrophage activation: applications for fish phagocytes. AB - The immune defence mechanisms of fish seem to be related and similarly competent to those of mammals. Because of this, there is an increased interest in the immune responses of fish as models for higher vertebrates in immunological/immunotoxicological studies. Macrophages (M phi), phagocytic cells of the mammalian and teleost immune system which reside in tissues, represent a quiescent population of cells. However, upon stimulation, alterations in the physiology of these resident M phi occur which can be defined in terms of activation. This study was undertaken to determine whether biological markers used to assess mammalian M phi activation are applicable for use with fish M phi. Cells were recovered from the peritoneal cavity of non-injected and Aeromonas salmonicida-injected fish, and differences between resident and elicited M phi were evaluated with respect to protein content, phagocytic competence, enzyme activities and hydrogen peroxide production. Results demonstrate that biological markers used to assess mammalian M phi activation, with the exception of acid phosphatase activity, can be used to characterize the activation state of trout M phi, and that the activation process in both fish and mammals may occur by similar mechanism(s). PMID- 8244467 TI - Intestinal epithelial cells express the CD23/Fc epsilon RII molecule: enhanced expression in enteropathies. AB - Immunohistochemical analysis of normal human intestine revealed that two anti CD23 monoclonal antibodies (mAb), EBVCS 1 and EBVCS 2, reacted with human intestinal epithelial cells. Both mAb exhibited an exclusive reactivity with epithelial cells of the small and large bowels. Staining with both EBVCS 1 and EBVCS 2 was localized on the apical and basal sides of enterocytes. Enhanced expression of CD23 on gut epithelial cells was found in inflammatory bowel diseases, in children with food intolerance to cows' milk proteins and in a young infant with severe autoimmune enteropathy. Western blot analysis of anti-CD23 mAb reactivity with gut epithelial cell extracts showed the presence of a non reducible 42,000-45,000 M(r) polypeptide compatible with the membrane form of the intact CD23 molecule. These data show that CD23 is constitutively expressed by intestinal epithelial cells and that its expression is enhanced in enteropathies. PMID- 8244468 TI - A new era for the Journal and for pediatrics in India. PMID- 8244469 TI - Status epilepticus in children. PMID- 8244470 TI - Mechanical ventilation in pediatrics. PMID- 8244471 TI - Ultrasonic evaluation of neonatal subependymal-intraventricular hemorrhage and its complications. AB - This study demonstrates the value of neurosonography in the evaluation of subependymal-intraventricular hemorrhage (SEH-IVH) and its major complications in the premature infant. Real time sector scans were performed on 150 premature neonates within first 24 hours of life (mean, 10 hrs), and then at 48, 72, and 96 hours and thereafter at weekly intervals. The incidence of SEH-IVH was 22% (n = 33). Twenty seven of the 33 infants had small hemorrhages (Grade I & II), 4 infants sustained Grade III hemorrhage and 2 infants had Grade IV hemorrhage. All hemorrhages occurred in the first 96 hours of life; 50% being diagnosed with the first scan. Larger grades of hemorrhages were generally symptomatic while majority of patients with small hemorrhages had no associated symptoms. The infants especially at risk were those less than 32 weeks' gestation (odds ratio, 29 fold) and/or birth-weight < 1200 gms (odds ratio, 6 fold). Three infants developed post hemorrhagic ventriculomegaly which subsided spontaneously in two of them by the age of 4 weeks. One infant progressed to post hemorrhagic hydrocephalus. The mortality in the study group was 11.3% (n = 17) and correlated to the severity of hemorrhage (39% vs 3.4%; hemorrhagic vs non hemorrhagic group, p < 0.001). At least one screening sonogram is essential within first 4 days of life to diagnose SEH-IVH, which may be otherwise clinically silent. Once the hemorrhage is diagnosed, followup evaluations may be done at weekly intervals to detect post-hemorrhagic ventriculomegaly. PMID- 8244472 TI - Gastrointestinal bleeding--diagnosis and management. AB - Gastrointestinal (GI) bleeding in infants and children provokes alarm in parents, and often anxiety in pediatricians. While this clinical situation has not changed over the years, advances in endoscopy and radiology, as well as new therapeutic modalities allow more accurate identification of the cause of bleeding and more effective treatment. Although the majority of GI bleeding in the pediatric population is self limiting, a systematic approach including prompt assessment, diagnosis, and treatment is necessary to preclude undue morbidity. PMID- 8244473 TI - Albendazole in neurocysticercosis. PMID- 8244474 TI - Ribavirin in clinical practice. PMID- 8244475 TI - Growth hormone therapy for Turner syndrome--is it indicated? PMID- 8244476 TI - Lipoblastomatosis of the neck. PMID- 8244477 TI - Parainfluenza virus type 3 and pertussis syndrome. PMID- 8244478 TI - Smith-Lemli-Opitz syndrome. PMID- 8244479 TI - Wenckebach phenomenon and motor aphasia in enteric fever. PMID- 8244480 TI - Infection by Dipylidium caninum through pet cats. PMID- 8244481 TI - Parenteral nutrition in the management of severe protracted diarrhea. AB - Seventeen children aged 3 weeks to 19 months with severe Protracted Diarrhea (PD), and who were deteriorating on our standard management protocol (including special diets) were given Parenteral Nutrition (PN) for 4 to 19 days with crystalline aminoacid solution (Vamin N) in 10% dextrose and lipid emulsion (Intralipid 10%). Peripheral lines were used in majority (84%). Enteral feeds were started early and rebuilt as per tolerance. The mean daily protein and caloric intake achieved by hyperalimentation was 2.2 +/- 0.7 g/kg and 106 +/- 41 K cal/kg respectively. Diarrheal control and improvement in nutritional status was achieved in all but 4 who died (2 of refractory diarrhea and 2 of sepsis, 1 of which was probably PN related). Other PN related, treatable complications included thrombophlebitis (11.8%), sepsis (17.6%), and metabolic imbalance (17.6%). PN solutions and accessories alone cost an approximate average of Rs. 280/day, with extras for biochemical monitoring (Rs. 70/day) and special nursing (Rs. 200/day). Only 5 of the 13 survivors had a significant relapse of PD, within 5 to 80 days of discharge, necessitating further PN in 2. There were no further deaths. PN was therefore, found to be of life saving value in 13 of 17 children with severe protracted diarrhea and therefore, must be available in specialised units caring for such children. PMID- 8244482 TI - Outbreak of neonatal septicemia with multidrug resistant Klebsiella pneumoniae. AB - A nosocomial outbreak of neonatal septicemia due to K. pneumoniae occurred in nursery during June-July, 1991. Klebsiella pneumoniae (Klebocin type 314) was recovered from blood of 33 (70.2%) of 47 neonates with septicemia. Multiple drug resistance was observed in all the cases. The same strain of K. pneumoniae was recovered from the neonates and environment of nursery and labour room as well. The outbreak was attributable to environmental dissemination. PMID- 8244483 TI - Efficacy of amphotericin B in multi-drug resistant kala-azar in children in first decade of life. AB - Fifty children in the first decade of life, and suffering from multiple drug resistant kala-azar, confirmed by demonstration of amastigotes in aspirates of bone marrow or spleen were treated with amphotericin B in gradually increasing dosage to a total dose of 20 mg/kg. All patients had classical features of severe kala-azar, and had taken more than one course of antimony and pentamidine, and three patients had taken one additional course of ketoconazole besides many courses of antimony and pentamidine. The clinical response started just after first infusion in 8 patients, and the patients became afebrile. By 5th infusion, all looked better and 18 patients became afebrile. By 15th infusion all patients were afebrile and cheerful. Their spleens became smaller and body weights and total white cell counts increased. Forty eight patients had parasitological cure at the end of treatment, and only 2 patients required an additional 5 infusions for parasitological cure. All patients were ultimately cured. No one relapsed within six months of follow up. All patients had shivering, rigor and rise of temperature on the day of infusion, which could be minimized with prior administration of low dose of hydrocortisone, but could not be eliminated. Eighteen patients had loose motions during treatment, while 14 patients had decrease in appetite which improved quickly when the treatment was over. Fourteen patients had transient rise of blood urea, in six patients serum creatinine also increased and 16 patients had a minor fall in serum potassium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244484 TI - Serodiagnosis of subacute sclerosing panencephalitis by enzyme linked immunosorbent assay. AB - Thirty three young patients (23 males and 10 females), between 3-20 years of age, with a clinical suspicion of subacute sclerosing panencephalitis (SSPE) were evaluated over a period of 4 years. A past history of measles was obtained in 42.4% of the cases. One child was immunised against measles. Measles specific antibodies (IgG) were demonstrated in the cerebrospinal fluid (CSF) from 30 (90.9%) of the 33 patients by ELISA. In the remaining three cases the antibodies were detected only in the serum. Serodiagnosis of SSPE by ELISA had a sensitivity of 100%, a specificity of 93.3% and a positive predictive value of 100%. PMID- 8244485 TI - Brain abscesses in cyanotic heart disease. AB - Sixty cases of brain abscesses in patients with cyanotic heart disease is reported. Forty four (73.3%) belonged to pediatric age group. There was only single case of infratentorial abscess located in the cerebellar paravermian region. The abscesses were multiloculated in 42%, multiple in 10% and solitary in 48% cases. Streptococci was the commonest (77%) micro-organism isolated from pus culture. Forty eight patients were treated by aspiration through twist drill, 7 patients conservatively and 5 had primary excision of abscesses. Forty four patients improved in their neurological status, three had fixed neurological deficits and 5 patients deteriorated further. There were 8 deaths. Complication like cyanotic spells was recorded in 17% cases treated by aspiration. None of the patients treated by twist drill aspiration had seizure. It is suggested that aspiration of abscesses through twist drill hole and antibiotic therapy could be the treatment of choice. Vascular slowing in a localised area resulting in infraction and thus preparing the focus for abscess formation is the possible mechanism in the evolution of these abscesses. PMID- 8244486 TI - The profile and outcome of patients admitted to a pediatric intensive care unit. AB - The records of all admissions to a 6-bed pediatric intensive care unit (PICU) over a period of 6 years were reviewed. The age, diagnosis, clinical service provided, duration of stay and outcome were recorded. Of the 3025 children admitted, 2092 (69.2%) were males. Neonates constituted 13.1% (400) and infants 57.1% (1727) of total admissions. The duration of stay ranged from 6 hours to 46 days, and 61 patients stayed for longer than 13 days (long-stay patients). The most common cause for admission was septicemia, seen in 459 patients (14.8%); 418 (13.8%) children had congenital heart disease, 407 (13.5%) lower respiratory tract infections (LRTI) and 261 (8.6%) meningitis. The most common conditions necessitating long-stay in the PICU were meningitis (20%), Landry-Guillain-Barre syndrome (16.6%), acute renal failure (20%), and septicemia (16.6%). There were 721 deaths giving a mortality of 23.5%. Of these 134 (18.6%) were due to septicemia, 103 (14.2%) due to congenital heart disease, 77 (10.6%) due to meningitis and 55 (7.6%) due to LRTI. The highest case fatality rate was seen with encephalitis (52.6%), followed by hepatic coma (51.3%), malignancies (43.2%), septicemia (29.1%) and meningitis (29.5%). The mortality was lower (9.8%) in long-stay patients than in short-stay patients (24.6%). There was gradual increase in proportion of cases requiring interventions including artificial ventilation (1% to 35%), peritoneal dialysis (1.5% to 11%), insertion of central venous pressure lines (0 to 10%), over the last 6 years.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244487 TI - Monitoring in pediatric intensive care. AB - Monitoring plays an essential role in the management of critically ill children, although continuous observation along with frequent clinical examination remains the best readily available monitor. Unfortunately, human beings do not have the capability of a prolonged, uninterrupted attention span and nurses often have multiple tasks assigned that limit their ability for continuous observation. Furthermore, some information cannot be obtained accurately by clinical examination alone, e.g. oxygen saturation, ICP, etc. Therefore, it is important to understand the principles and practical points to be able to use these monitors, and first rule out malfunction, disconnection, or improper electrode placement when abnormal reading appear on these monitors before initiating clinical intervention on a patient. PMID- 8244488 TI - Procedures in pediatric intensive care unit. PMID- 8244489 TI - Multiple trauma--stabilization and management in pediatric intensive care unit. AB - Unlike many medical problems which allow us the luxury of casual contemplation or a diligent search for relevant references for assistance, the care of children suffering major trauma does neither. Our only proper response to these young patients is a rapid assessment and treatment, often performed simultaneously, if we are to preserve life and limb. The anxiety this challenge presents us cannot be minimized: the almost unbelievably thin margin for error in injured youngsters, their small total blood volume, the variation in blood pressure, other vital signs and drug dosages with age, and the family's justifiable hope for a complete recovery all account, in part, for this anxiety. It is, however, precisely for these reasons that our preparation and knowledge is critical if the disability and mortality too often associated with childhood trauma are to be circumvented. Neither the price of failure nor the rewards of success can be measured--they're both too high. PMID- 8244490 TI - Electrolyte problems in critically ill children. PMID- 8244491 TI - Mycoplasma infections of the human respiratory tract. AB - The knowledge on the role of M. pneumoniae in human respiratory tract infections especially in infants and children has advanced in the past two decades. The understanding of its incidence, clinical presentations, pathology & pathogenesis, laboratory diagnosis, complications and late sequelae, treatment, prevention and epidemiological aspects place it as an important aetiological agent in the pediatric patients. It should be considered in the differential diagnosis of all pneumonias. Further studies are needed to solve many observations on the role of mycoplasma in diseases of man. PMID- 8244492 TI - Von Willebrand's disease. PMID- 8244493 TI - Refractory anaemias. PMID- 8244494 TI - Oncogenes: present status. AB - Oncogenes are genes associated with causation of cancer. They were originally associated with the ability of retroviruses to cause tumors in animals. These viral oncogenes (V-onc) have their cellular counterparts (C-onc) called Proto oncogenes. Function of Proto oncogenes is to maintain cellular growth and development. Activation of these proto-oncogenes can occur due to mutation which leads to uncontrolled cell growth. The Proto oncogenes can be grouped into different categories based on their protein products, i.e. protein kinases, growth factors, growth factor receptors, and DNA binding proteins. There are also genes that normally suppress malignant transformation and these are called anti oncogenes. Loss of their suppressor activity leads to unimpeded growth. Oncogene abnormalities are seen in pediatric leukemias, lymphomas, and various solid tumors. Anti oncogenes are associated with retinoblastoma (Rb gene), Wilms' tumor, rhabdomyosarcoma and neuroblastoma, etc. Identification of these abnormalities have diagnostic, prognostic and therapeutic implications. The utility of oncogenes in classification of human cancer and monitoring cancer therapy is quite clear, but the future of these for therapeutic interventions remains uncertain. Role of c-abl oncogene in chronic myeloid leukemia (CML), bcl 2, in lymphomas, N-myc in neuroblastomas and retinoblastoma (Rb) gene in retinoblastomas is well understood and used in designing proper therapeutic approaches. Since oncogenes also control normal cellular function, their use for therapy may be limited by the amount of damage to normal cells. Their maximum therapeutic benefit may be realized only when used in combination with other modalities. PMID- 8244495 TI - Cytogenetics in childhood leukemias and malignant lymphomas. PMID- 8244496 TI - Supportive therapy in management of leukemias. PMID- 8244497 TI - Retinoblastoma: a review. AB - Retinoblastoma is the most common primary ocular malignancy in childhood, usually occurring below the age of five. Recent advances in molecular biology have enabled understanding of retinoblastoma tumorigenesis as well as detecting carriers of the mutant retinoblastoma allele. It is possible now to predict prenatally whether a child carries the retinoblastoma genome. This has enabled early detection of retinoblastoma and improved outcome. Also these children are at risk for mesenchymal tumors in early adult life necessitating life long vigilance. Great strides have been made in the management of retinoblastoma. Introduction of a new pathological staging system and addition of combination chemotherapy for extraocular disease has led to more than 80% long term disease free survivors in this group of patients, who earlier had a very poor outcome. For localized intraocular disease, local treatment seems to be all that is necessary. Newer radiation techniques have helped preserve useful vision and reduced radiation related side effects. PMID- 8244498 TI - Perspectives in the management of central nervous system tumours. AB - A general review and update in the management of CNS tumours using medulloblastoma as the main model is given in this article. Special emphasis has been placed on the benefits of combined modality treatment for brain tumours. The pathogenesis and management of these tumours is discussed and recommendations made for treatment in developing countries. PMID- 8244499 TI - Childhood non-Hodgkins lymphoma. PMID- 8244500 TI - The importance of pulmonary function tests in adenotonsillectomy indications. AB - Children who had undergone adenotonsillectomy for recurrent adenotonsillitis showing no signs of clinical or radiological obstructive manifestations were evaluated with pulmonary function tests before, and one month after the operation. In relation to the result obtained by function tests, 60% of 45 cases (27) had the findings of mild obstructive pulmonary disease whereby these findings were in transient character that vanished after the operation. The following parameters were measured and found that they were all increased, mean FVC from 82.22 +/- 6.82 to 93.11 +/- 7.81 (p < 0.01), mean PEF from 77.60 +/- 8.38 to 88.60 +/- 5.57 (P < 0.01), mean FEVI from 74.28 +/- 11.68 to 90.15 +/- 7.28 (p < 0.01), mean FEF 25 from 71.44 +/- 11.53 to 83.53 +/- 6.40 (p < 0.01), mean FEF 50 from 69.53 +/- 14.53 to 84.37 +/- 7.72 (p < 0.01), mean FEF 75 from 70.08 +/- 12.15 to 85.48 +/- 7.15 (p < 0.01). In conclusion, pulmonary function tests could reveal the obstructive effects of adenotonsillar hypertrophy with no clinical or radiological obstructive findings, and could be useful in surgical indications of adenotonsillar hypertrophy dur to recurrent infections in children. PMID- 8244501 TI - Cord blood diazepam: clinical effects in neonates of eclamptic mothers. AB - Diazepam used in the treatment of eclampsia crosses the placental barrier readily, and may cause various clinical effects in the neonates. Twenty-five (25) live born babies of eclamptic mothers receiving diazepam were studied and cord blood diazepam concentration was estimated. Effect of low dose of diazepam is minimal apart from lowering of rectal temperature and the effects lasted for a period of 12 hours. But high dose (> 30 mg) of diazepam and prolonged duration of diazepam therapy in mothers causes significant depression of the newborn and the effects lasted for a period of 36-48 hours. As the clinical condition of the newborn is not related to the diazepam concentration in cord blood, the cord blood estimation is not helpful in the assessment of clinical effects of the drug in newborn. The tissue storage of the drug in newborn appears to be responsible for the clinical effects. PMID- 8244502 TI - Nerve conduction studies and its importance in diagnosis of acute poliomyelitis. AB - Motor nerve conduction velocities was performed on 50 subjects in the pediatric age group. Thirty two patients with acute poliomyelitis and 18 controls. The MNCV was studied in the median nerve in the upper limb and the posterior tibial in the lower limb. The motor nerve conduction velocity in polio patients matched well with the controls, as well as within the accepted standards for normal. The MNCV of the median nerve ranged from 41.8 +/- 2.76 m/sec in under 1 year to 44 +/- 2.1 m/sec in 3-8 years, in polio patients, while the range in controls varied from 37 to 53 m/sec. Similarly, for the posterior tibial nerve, in polio patients the value of MNCV varied from 38.7 +/- 4.9 m/sec to 42.5 +/- 3.1 m/sec. In the controls, also the MNCV ranged from 38.5 +/- 6.3 m/sec to 48.4 +/- 3.42 m/sec. Thus, no delay on the motor nerve conduction velocity was seen. Poliomyelitis is a major problem in developing countries like Pakistan and India, where serological diagnosis is a luxury. The determination of motor nerve conduction velocity provides a quick and easy method of distinguishing poliomyelitis from other motor nerve disorders esp. Guillain Barre syndrome. PMID- 8244503 TI - Maternal risk factors associated with low birth weight. AB - Maternal factors comprising of social, obstetric and anthropometric are found to influence LBW. The present study had found association between obstetric risk factors like age of the mother, parity and gravida with LBW. Similar association was also observed between maternal height, and maternal weight with LBW. However, social factors were not found to be associated with LBW. This could probably be due to RUHSA's intervention which requires a further inquiry. PMID- 8244504 TI - Oral polio vaccination in infants: beneficial effect of additional dose at birth. AB - This study was done to assess the response of newborns to trivalent oral polio vaccine and to study any efficacy of OPV if given to infants on third day of life. The study was conducted in two groups, A (87) and B (55) of infants in Delhi, India. In group A, the children received one birth dose or 'O' dose of TOPV, followed by 3 conventional doses started at 6 weeks, and in group B the children received only 3 doses of OPV. Pre and one month post immunization serum samples were tested for the presence of neutralising antibodies. In addition, in group A serum samples were collected at 6 weeks before the administration of 1st dose to see the sero response following 'O' dose of TOPV. It was found that administration of OPV on 3rd day of life leads to sero conversion in 15.3% of infants to all three polio virus types by the age of 6 weeks, and highest sero response was seen for polio virus type 1. Sero-conversion in group A was significantly more than sero-conversion in group B after the administration of last dose. Thus the study has established that immunization of newborns with TOPV is a safe and effective means for improving protection against the disease. PMID- 8244505 TI - Management of ARI for control of mortality in underfives. AB - WHO advocated a standard case management for control of ARI related mortality. India has started an ARI control programme in 15 districts. Conflicting reports appeared regarding the effectiveness of case management strategy. Authors reported earlier that it was not effective. The coverage was only about 57% in pneumonias, and 23% in severe pneumonias. Health agencies & health workers have a low utilization (13.7%), and RMPs were used in large numbers (36.9%). Most of the deaths were in those treated by RMPs (75%). The efficacy of standard case management (88%) is highlighted. The problem of coverage and assumptions behind the strategy were discussed and recommendations for improvement were suggested. PMID- 8244506 TI - Coxsackie virus infections in rheumatic fever. AB - The close relationship between Group A beta Hemolytic Streptococci (GABS) and rheumatic fever is a well established one. However, the concept of the streptococcus as the sole etiologic agent of the rheumatic heart disease (RHD) has been challenged over the past years. Since coxsackievirus group B (CVB) has long been proposed as a cause of acquired valvular disease simulating rheumatic fever, we attempted in this study to document infections with this group of viruses in patients with rheumatic fever. We obtained blood samples from 106 patients with old (quiescent) rheumatic fever/rheumatic heart disease [group I], 94 patients with acute rheumatic fever (ARF) [group II], and 74 normal matched controls. We tested for the presence of neutralizing antibodies to the 6 serotypes of CVB by a micro neutralization test. We have found that infection with CVB, especially types B2 and B4, was common in the studied population. Forty two percent of normal individuals had evidence of infection with any of the 6 serotypes of CVB. Patients of group I had significantly more frequent infections with CVB 2. Patients in group II had significantly more frequent infections with CVB 2 and CVB 6. There was no clear correlation between such infections and the clinical course of rheumatic fever. There was no difference in the incidence of CVB infections between patients with definite ARF, and patients with suspected ARF. We set a low order association between rheumatic fever and infection with CVB types B2 and B6. We emphasize the importance of pursuing the investigation of the role of CVB in relation to RHD. PMID- 8244507 TI - Transient Isospora belli infection in a normal child. PMID- 8244508 TI - Fibrous dysplasia in two siblings. PMID- 8244509 TI - Cole-Carpenter's syndrome. PMID- 8244510 TI - Tricho-rhino-phalangeal syndrome type-1. PMID- 8244511 TI - Bone marrow involvement in embryonal rhabdomyosarcoma. PMID- 8244512 TI - Normotensive blacks have heightened sympathetic response to cold pressor test. AB - The purpose of this study was to compare sympathetic nerve activity responses to the cold pressor test in black and white normotensive subjects. We recorded muscle sympathetic nerve activity (microneurography of the peroneal nerve), arterial blood pressure, and heart rate in 9 normotensive American blacks (24 +/- 2 years, mean +/- SEM) and 10 normotensive American whites (28 +/- 2 years) at rest and during hand immersion in ice water (cold pressor test). Body weight was not different in the two groups (72.4 +/- 3.7 versus 74.1 +/- 3.8 kg, black versus white subjects). During supine rest, mean arterial pressure (92 +/- 2 versus 93 +/- 3 mm Hg, black versus white), heart rate (66 +/- 4 versus 62 +/- 3 beats per minute, black versus white), and muscle sympathetic nerve burst frequency (12 +/- 2 versus 17 +/- 3 bursts per minute, black versus white) were not different in the two groups. During the cold pressor test, mean arterial pressure, heart rate, and muscle sympathetic nerve activity increased from supine rest in both groups. The magnitudes of increases in mean arterial pressure and total minute muscle sympathetic nerve activity were significantly greater in blacks than whites (33.5 +/- 3 versus 22.4 +/- 3 mm Hg and 416 +/- 24% versus 243 +/- 31% of control, respectively, black versus white, P < .05). The increases in heart rate were most significantly different for the two groups. These data suggest that the enhanced pressor response to cold stress observed in normotensive blacks is attributable to greater increases in peripheral sympathetic nerve activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244513 TI - Effect of captopril on vasoconstriction and Ca2+ fluxes in aortic smooth muscle. AB - The effects of captopril on the response of cytosolic free Ca2+ concentration in cultured vascular smooth muscle cells of aortas from Wistar-Kyoto and spontaneously hypertensive rats to angiotensin II (Ang II) and bradykinin were studied using fura 2. Incubation with captopril for longer than 10 minutes caused a decreased response of cytosolic free Ca2+ to Ang II and bradykinin. Maximal effects of captopril were observed after a 40-minute incubation. The inhibitory effect of captopril was abolished in Ca(2+)-free medium, suggesting that captopril acts by blocking Ca2+ influx. Similar effects were observed with enalaprilat. Isometric contraction of aortic strips induced by Ang II in normotensive rats was reduced from 6.5 +/- 2.5 to 1.8 +/- 0.6 mN by a 40-minute incubation with 1 mumol/L captopril (P = .016). Enalaprilat similarly decreased the Ang II-induced contraction. Besides the inhibition of the angiotensin converting enzyme, direct effects of Ang II converting enzyme inhibitors on vascular contraction and Ca2+ influx in vascular smooth muscle cells may be of therapeutic relevance. PMID- 8244514 TI - Is white coat hypertension innocent? Structure and function of the heart in the elderly. AB - To evaluate the morphological and functional characteristics of the heart in elderly patients with white coat hypertension, we performed an echocardiographic study in 67 elderly individuals older than 60 years: 17 patients with white coat hypertension, 34 patients with true hypertension, and 16 normotensive control subjects. White coat hypertension was defined as a mean 24-hour ambulatory systolic blood pressure of less than 140 mm Hg associated with office hypertension. Cardiac responses to an isometric handgrip exercise test were used to evaluate left ventricular functional reserve. Left atrial dimension and left ventricular mass index were significantly greater in the white coat hypertension group than in the normotension group (P < .05) but were similar to values in the true hypertension group. Left ventricular diastolic function, expressed by peak late-early filling ratio of diastolic mitral flow, showed increasing impairment in the order of the normotension, white coat hypertension, and true hypertension groups (analysis of variance, P < .05); the ratio in the white coat hypertension group tended to be higher than that in the normotension group (unpaired t test, P = .054). The relation between fractional shortening and end-systolic stress did not shift downward after handgrip exercise in the white coat hypertension group, indicating that functional reserve in the left ventricle was maintained. Thus, patients with white coat hypertension had a moderately increased left atrial dimension and left ventricular mass in association with a tendency for disturbed diastolic function, although systolic functional reserve remained the same. These findings suggest that white coat hypertension in the elderly may not be innocent. PMID- 8244515 TI - Arterial hemodynamics in human hypertension. Effects of angiotensin converting enzyme inhibition. AB - Previous studies have shown some distinct hemodynamic alterations in essential hypertension, including increased resistance, wave reflections, and pulse wave velocity and decreased systemic compliance. These abnormalities are completely normalized by nonspecific smooth muscle dilation with nitroprusside but not by combined alpha- and beta-adrenergic blockade. The renin-angiotensin system, acting possibly via both circulating and local tissue effects, is thought to play an important role in essential hypertension, so its role in the altered hemodynamics deserves careful investigation. A hypertensive patient group was compared with a normotensive group similar in age, body size, and proportion of men and women. During diagnostic cardiac catheterization, ascending aortic micromanometer pressures and electromagnetic flows were measured in the baseline state. Intravenous captopril of a sufficient dosage (11 mg) to normalize blood pressure then was given to the hypertensive patients while measurements were repeated. From the pressures and flows, aortic input impedance, wave reflection magnitude, and compliance were computed. In the hypertensive group, the important hemodynamic alterations consisted of increased peripheral resistance, first zero crossing of aortic impedance phase angle, and wave reflections and decreased systemic compliance. Captopril had a pronounced hemodynamic effect. It normalized blood pressure, resistance, and impedance phase angle zero crossing. Compliance, although increased substantially by captopril, was still slightly lower than normotensive levels. The magnitude of wave reflections, although substantially lowered by angiotensin converting enzyme inhibition, was still persistently greater than normal. The present results, together with those previously reported, demonstrate that a complex interplay of factors underlies the increased smooth muscle tone in essential hypertension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244516 TI - Caffeine attenuates the renal vascular response to angiotensin II infusion. AB - Non-modulation has been proposed as an intermediate phenotype in human essential hypertension. The trait is characterized by blunted aldosterone and renal plasma flow responses to short-term angiotensin II (Ang II) infusion. Elevated tissue Ang II levels or decreased tissue adenosine levels could account for this decreased sensitivity to Ang II. In support of the latter possibility, endogenous adenosine has been shown to contribute to the renal vasoconstrictive response to Ang II in animals. We therefore tested the hypothesis that endogenous adenosine contributes to modulation of renal plasma flow in sodium-replete humans. We examined the effect of long-term administration of the adenosine receptor antagonist caffeine on baseline renal plasma flow and on the renal plasma flow response to short-term Ang II infusion in six salt-replete normotensive subjects in a single-blind, placebo-controlled study. para-Aminohippurate clearance was used to assess renal plasma flow. Ang II was infused in graded doses (0.3 to 3 ng/kg per minute) in the presence and absence of caffeine (250 mg PO TID for 7 days). Blood pressure, plasma renin activity, Ang II, electrolytes, and para aminohippurate clearance were measured before and after each dose of Ang II. Caffeine did not alter either baseline blood pressure or the blood pressure response to Ang II but did increase baseline plasma renin activity from 0.72 +/- 0.09 to 1.42 +/- 0.26 ng angiotensin I/mL per hour (P = .01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244517 TI - Effects of angiotensin II and endothelin-1 on platelet aggregation and cytosolic pH and free Ca2+ concentrations in essential hypertension. AB - The aims of this study were to determine the relations between platelet free calcium concentrations ([Ca2+]i), intracellular pH (pHi), and aggregation and to assess the effects of angiotensin II (Ang II) and endothelin-1 on these platelet parameters in normotensive subjects and hypertensive patients. Seventeen normotensive subjects, 25 untreated hypertensive patients, and 34 treated hypertensive patients were studied. Platelet cytosolic free [Ca2+]i and pHi were measured spectrofluorometrically using specific fluorescent probes (fura 2-AM and BCECF-AM, respectively) in unstimulated and Ang II- and endothelin-1-stimulated platelets. Aggregation was measured by a turbidometric technique. Basal [Ca2+]i (141 +/- 11 nmol/L) and pH (7.16 +/- 0.01) were higher (P < .05) in the untreated hypertensive group compared with the normotensive (118 +/- 9 nmol/L, 7.11 +/- 0.01, respectively) and treated hypertensive (121 +/- 11 nmol/L, 7.12 +/- 0.01, respectively) groups. In the combined normotensive and hypertensive groups, there were significant correlations between [Ca2+]i and mean arterial pressure (r = .75, P < .01), pHi and mean arterial pressure (r = .72, P < .01), [Ca2+]i and pHi (r = .71, P < .01), [Ca2+]i and aggregation (r = .69, P < .02), and pHi and aggregation (r = .56, P < .05). Ang II stimulation significantly increased [Ca2+]i and pHi in the untreated hypertensive and normotensive groups. The net change in [Ca2+]i induced by Ang II was significantly higher (P < .05) in the untreated hypertensive group compared with the other groups (67 +/- 6 nmol/L for the untreated hypertensive group versus 54 +/- 5 and 29 +/- 8 nmol/L for the normotensive and treated hypertensive groups, respectively). In the presence of Ang II, thrombin-induced aggregatory responses were increased in all three groups, but the maximal response was significantly higher in the untreated hypertensive group compared with the other groups (P < .05). Endothelin-1 increased pHi through endothelin A-receptors (effect blocked by the specific antagonist BQ-123) but had no significant effect on [Ca2+]i or aggregation. However, endothelin-1 blunted thrombin-induced platelet aggregation in normotensive subjects but not in hypertensive patients. In conclusion, increased Ang II-stimulated [Ca2+]i and pHi in platelets of essential hypertensive patients may be associated with increased aggregatory responses. The stimulatory effect of endothelin-1 on pHi but not on [Ca2+]i or aggregation suggests that in platelets endothelin-induced signaling pathways other than phospholipase C may be involved.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8244518 TI - L-arginine administration normalizes pressure natriuresis in hypertensive Dahl rats. AB - A blunted pressure-natriuretic response characterizes hypertension in the Dahl salt-sensitive rat. Long-term L-arginine administration prevents hypertension in these animals. To determine if long-term L-arginine corrects the pressure natriuretic response, we gave salt-sensitive rats on an 8% sodium diet L-arginine or vehicle daily for 3 weeks. Identically treated salt-resistant rats served as controls. After 3 weeks, acute pressure-natriuresis curves were determined. To control for hypertension-induced renal damage, we also examined pressure natriuresis in salt-sensitive rats after short-term L-arginine. Baseline mean arterial pressure was 158 +/- 3 mm Hg in vehicle-treated salt-sensitive rats and 127 +/- 3 mm Hg in chronically L-arginine-treated salt-sensitive rats. During alterations in perfusion pressure, renal blood flow was autoregulated in all groups. Glomerular filtration rate was autoregulated in salt-resistant rats and L arginine-treated salt-sensitive rats but fell with decreasing pressure in vehicle treated salt-sensitive rats. Sodium excretion was greater (P < .05) in L-arginine treated than in vehicle-treated salt-sensitive rats and did not differ from salt resistant rats at 100, 125, and 158 mm Hg. The slope of the pressure-natriuresis relation was greater (P < .05) in chronically L-arginine-treated than in vehicle treated salt-sensitive rats. L-Arginine had no effect on natriuresis in salt resistant rats. Thus, long-term L-arginine administration normalizes pressure natriuretic responses in salt-sensitive rats. The effect is not due to the prevention of renal damage and is specific to the salt-sensitive strain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244519 TI - Pregnancy restores the renal vasodilator response to glycine in Dahl salt sensitive rats. AB - To determine if pregnancy alters the impaired renal vasodilator responses in hypertensive Dahl salt-sensitive (Dahl S) rats, we measured glomerular filtration rate and effective renal plasma flow and calculated renal vascular resistance before, during, and after renal vasodilation with glycine (0.17 mmol/kg per minute IV). Conscious, midterm (day 11 to 14) pregnant and age-matched virgin Dahl S and Dahl salt-resistant (Dahl R) rats were fed an 8% NaCl diet for 4 weeks (n = 6 per group). Mean arterial pressure was elevated (P < .05) in Dahl S compared with Dahl R rats, with no significant difference between pregnant and virgin animals in either group. Pregnancy resulted in significant increases in plasma volume, baseline glomerular filtration rate, and renal plasma flow and a significant decrease in renal vascular resistance. The glycine-induced increase in filtration rate in virgin Dahl S rats (27 +/- 4%) was less (P < or = .01) than pregnant Dahl S (60 +/- 4%) and either group of Dahl R (virgin, 43 +/- 3%; pregnant, 45 +/- 5%) rats. Similarly, the 21 +/- 4% increase in renal plasma flow in virgin Dahl S rats was less (P < or = .01) than the pregnant Dahl S (45 +/- 4%) and either pregnant (45 +/- 5%) or virgin (45 +/- 5%) Dahl R rats. Glycine decreased renal vascular resistance only 12 +/- 2% in the virgin Dahl S compared with 31 +/- 3% in the pregnant Dahl S rats and 30 +/- 4% and 28 +/- 3% in pregnant and virgin Dahl R rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244520 TI - Wave reflections and cardiac hypertrophy in chronic uremia. Influence of body size. AB - Arterial wave reflections (AWRs), an important determinant of cardiac afterload, are increased in hemodialysis patients. However, an association between AWRs and left ventricular hypertrophy has not been established in these subjects. We therefore sought an association between these two parameters in a cross-sectional study of two outpatient hemodialysis populations. AWRs were quantified (augmentation index, percent) as the ratio of the height of the late systolic peak to the total height of the carotid pulse wave recorded with a micromanometer tipped probe. AWRs were determined in 44 hemodialysis patients, 22 with pronounced AWRs (group A: augmentation index > 12%), and 22 with small or moderate AWRs (group B: augmentation index < 12%). The groups were matched for age, sex, blood pressure, and hemodialysis duration. Left ventricular size was determined by echocardiography. Despite lower body size (P < .005) and similar blood pressure, cardiac output, peripheral resistance, and aortic pulse wave velocity, group A patients had greater left ventricular mass (P < .01). For the population as a whole, left ventricular mass and AWRs were positively correlated (P < .0001) independent of age, blood pressure, hemodialysis duration, or body size. Lesser body height was the principal factor associated with increased AWRs in group A (P < .001). We conclude that in hemodialysis patients AWRs are associated with the development of left ventricular hypertrophy and that small body height is a risk factor for long-term cardiovascular complications. PMID- 8244521 TI - Renal acid-base excretion in normotensive salt-sensitive humans. AB - Reduced extracellular pH and bicarbonate levels recently have been reported in normotensive salt-sensitive subjects. To assess the possible role of altered renal acid-base handling in the perturbation of acid-base status in these individuals, we measured the renal acid-base excretion after an acute oral administration of either an alkali or acid load in normotensive salt-sensitive and salt-resistant men. Twenty-four young (22 to 29 years old), healthy male volunteers were placed on a low-salt diet (20 mmol NaCl per day) for 2 weeks with either 220 mmol NaCl or placebo added to the low-salt diet for 1 week each in a randomized single-blind crossover order. Salt sensitivity was defined as a significant drop in mean arterial pressure (> 3 mm Hg, mean of 60 readings taken on the seventh day of each diet, P < .05) during the low-salt diet. On the fifth and seventh days of each week, subjects were given an oral load of either sodium citrate (0.7 mmol/kg) or ammonium chloride (2.2 mmol/kg), respectively, in a randomized order, and arterial and urinary acid-base status was assessed at baseline and followed for 8 hours thereafter. According to the above definition, 13 subjects were considered salt sensitive. During the high-salt diet, mean arterial pressure was higher in the salt-sensitive than in the salt-resistant group (P < .01). Cumulative urinary bicarbonate excretion after the administration of sodium citrate was lower in the salt-sensitive than in the salt resistant subjects during both the low-salt (46%, P < .001) and high-salt (32%, P < .01) diets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244522 TI - Cardiovascular reactivity, coronary risk factors, and sympathetic activity in young men. AB - We have previously demonstrated that awareness of high blood pressure may increase blood pressure, plasma catecholamine levels, and stress responses. In the present study, three groups of 19-year-old men, all unaware of their blood pressure status, were selected from the first (group-1, 62 +/- 2 mm Hg, [mean +/- SEM], n = 15), 50th (group-50, 90 +/- 4 mm Hg, n = 15), and 99th (group-99, 123 +/- 5 mm Hg, n = 14) percentiles in causal mean blood pressure at a screening. They were studied (blinded examiners) with intra-arterial blood pressure recordings and multiple measurements of arterial plasma epinephrine and norepinephrine during a mental arithmetic challenge and cold pressor test. Despite high mean blood pressure at the screening, group-99 did not differ from group-50 either in intra-arterial mean blood pressure after 30 minutes of supine rest (89 +/- 3 versus 86 +/- 2 mm Hg) or in serum lipids and resting plasma epinephrine and norepinephrine. However, in group-99 resting plasma epinephrine showed a positive hyperbolic relation to resting diastolic blood pressure (r = .73, P = .004) and a negative hyperbolic relation to the ratio of high-density lipoprotein cholesterol to total cholesterol (r = -.75, P = .002). None of these correlations were present in the two other groups. Furthermore, the three groups differed in heart rate responses (P < .0005) and systolic (P < .0005) and diastolic (P < .05) blood pressure responses to mental arithmetic challenge, group-99 being hyperreactive compared with the other two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244523 TI - Ambulatory blood pressure of adults in Ohasama, Japan. AB - We performed a cross-sectional study in a small town in northern Japan to evaluate the distribution, reference values, and daily variation in ambulatory blood pressure. A total of 705 subjects (229 men aged 61.3 +/- 13.4 years [mean +/- SD] and 476 women aged 57.5 +/- 13.3 years; 41.1% of the regional adult population, n = 1716), including those treated with antihypertensive drugs (n = 231, 66.5 +/- 9.5 years) as well as untreated subjects (n = 474, 55.0 +/- 13.5 years), participated in the study. Both ambulatory and screening blood pressures were measured in 659 subjects. Ambulatory blood pressure was measured with an automatic device (Colin ABPM-630). The 24-hour ambulatory blood pressure in the total population was 121.7 +/- 13.0/71.1 +/- 7.6 mm Hg (95th percentile value [95%] = 146/85 mm Hg). The corresponding value in the untreated subjects was 119.4 +/- 12.5/70.1 +/- 7.4 mm Hg (95% = 144/83 mm Hg). The 24-hour average ambulatory blood pressure was 118.0 +/- 11.1/69.4 +/- 6.8 mm Hg (95% = 139/81 mm Hg) in subjects identified as normotensive by their screening blood pressure (n = 448, 57.2 +/- 13.1 years) and 133.6 +/- 14.2/78.9 +/- 8.8 mm Hg in those identified as hypertensive by their screening blood pressure (n = 73, 63.1 +/- 10.6 years). Based on the mean+SD of the 24-hour ambulatory blood pressure in the normotensive subjects by their screening blood pressure (129/76 mm Hg), the 24 hour ambulatory blood pressures in 25 (34.2%) of these 73 hypertensive subjects by screening blood pressure were below this level. Nine (2%) of 448 normotensive subjects by screening blood pressure were above the mean+2 SDs (140/83 mm Hg) of the 24-hour ambulatory blood pressure in the normotensive group by screening blood pressure. Ambulatory and screening blood pressures increased with age. The age-dependent increase in ambulatory blood pressure was less apparent in men. The 24-hour average pulse rate decreased with age. The daily variation in ambulatory blood pressure (standard deviation) increased with age, whereas that of pulse rate decreased with age. Increases in blood pressure variation were observed in nighttime and daytime blood pressure values. The differences between day versus night ambulatory blood pressures decreased with age in men but not in women.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8244524 TI - Endothelial dysfunction augments myogenic arteriolar constriction in hypertension. AB - To elucidate the underlying reason or reasons for the increased peripheral resistance in hypertension, we investigated the pressure-diameter relation--the myogenic response--of isolated, cannulated arterioles (approximately 50 microns) of cremaster muscle of 12-week-old Wistar-Kyoto (WKY) rats, spontaneously hypertensive rats (SHR), and normal Wistar (NW) rats. All arterioles constricted in response to step increases in perfusion pressure from 20 to 160 mm Hg. This constriction was, however, significantly enhanced from 60 to 160 mm Hg in arterioles of SHR compared with NW or WKY rats. For example, at 80 and 140 mm Hg, respectively, the normalized diameter (expressed as a percentage of the corresponding passive diameter of arterioles of SHR) was 11.8% and 27.6% (P < .05) less compared with those of WKY rats. Endothelium removal eliminated the enhanced pressure-induced tone in SHR. Similarly, indomethacin (10(-5) mol/L, sufficient to block prostaglandin synthesis) or SQ 29,548 (10(-6) mol/L), a thromboxane A2-prostaglandin H2 receptor blocker that inhibited vasoconstriction to the thromboxane agonist U46619, attenuated the enhanced pressure-diameter curve and reversed the blunted dilation to arachidonic acid in SHR. In contrast, the thromboxane A2 synthesis inhibitor CGS 13,080 (5 x 10(-6) mol/L) did not affect the increased pressure-induced tone or the reduced dilation to arachidonic acid in SHR. Thus, the present findings suggest that in early hypertension pressure-induced arteriolar constriction is increased. This seems to be due to an enhanced production of endothelium-derived constrictor factors, primarily prostaglandin H2. PMID- 8244525 TI - A gamma-melanocyte stimulating hormone and postnephrectomy natriuresis. PMID- 8244526 TI - Development of radial glia and astrocytes in the spinal cord of the North American opossum (Didelphis virginiana): an immunohistochemical study using anti vimentin and anti-glial fibrillary acidic protein. AB - We have shown previously that rubrospinal axons grow around a lesion of their pathway in developing opossums and that a critical period exists for that plasticity. As a first step toward addressing the possibility that glial maturation and/or the development of an astrocytic response to lesioning contribute to loss of rubrospinal plasticity, we have studied the normal development of radial glia and astrocytes in the spinal cord of the opossum by immunostaining for vimentin (Vim) and glial fibrillary acidic protein (GFAP). Vim like immunoreactivity (Vim-LI) was present in radial glia throughout the spinal cord at birth (12 days after conception), whereas GFAP-like immunoreactivity (GFAP-LI) was limited to cells of comparable morphology in the ventral part of the cervical cord. The subsequent appearance of GFAP-LI followed ventral to dorsal and rostral to caudal gradients and by postnatal day (PD) 15, it was found in radial glia throughout the cord. At the same age, processes immunostained by either antibody had lost their radial orientation in the ventral horn of the cervical cord. The subsequent transformation from radial glia to astrocytes also followed ventral to dorsal and rostral to caudal gradients. By PD30, mature appearing astrocytes were immunostained by both antibodies at thoracic levels of the spinal cord, the levels lesioned in the plasticity experiments referred to above, and by PD41, they were found at all levels of the cord.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244527 TI - Development of an astrocytic response to lesions of the spinal cord in the North American opossum: an immunohistochemical study using anti-glial fibrillary acidic protein. AB - We have shown previously that rubral axons grow around a lesion of their pathway in developing opossums and that a critical period exists for that plasticity. The critical period begins when rubral axons first reach the level of the lesion and ends sometime between postnatal days (PD) 26 and 30. The aim of the present study was to examine the development of an astrocytic response to lesioning the spinal cord to determine if there is a temporal correlation between the development of such a response and the end of the critical period. The astrocytic response was examined immunohistochemically, 2 and 4 weeks after hemisecting the thoracic spinal cord, using an antibody to glial fibrillary acidic protein (GFAP). A response was first seen at PD21 in the 2-week series. The response was relatively mild, however, and limited to the white matter. When the lesion was made at PD26, the response was still restricted to the white matter, but hypertrophied astrocytes were found at the gray/white matter junction and cystic cavities were present. When the lesion was made at PD41, the response had spread to the gray matter and it occupied a larger area rostral and caudal to the lesion than at earlier ages. The animals allowed to survive 4 weeks after lesioning were subjected to a second operation 4-5 days before sacrifice so that Fast Blue could be injected bilaterally two to three segments caudal to the lesion. When the hemisection was made at PD15, a response was present in the ventral and ventrolateral funiculi, but not in that part of the lateral funiculus that contains rubrospinal axons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244528 TI - Native astrocytes do not migrate de novo or after local trauma. AB - While transplanted astrocytes migrate in specific patterns in the recipient brains, it is not known whether native astrocytes behave similarly. The ability of normal astrocytes to migrate under non-transplant conditions was therefore explored. Native astrocytes were labelled in situ with fluorescent latex beads. These latex spheres were actively endocytosed by astrocytes in vitro, and it was therefore anticipated that these spheres would also be endocytosed by native astrocytes exposed to them. Labelling was accomplished by dissecting the pia mater away from a small region of the cerebral cortex and overlaying the area with Gelfoam containing fluorescent beads. After 2-4 h, the Gelfoam was removed and the wound was closed. At the end of 2-4 weeks, manipulated brains were harvested for fluorescence microscopy. In this analysis, fluorescent polyspheres had been taken up by both pial fibroblasts and astrocytes at the pial-glial margin. Labelled astrocytes [identified by glial fibrillary acidic protein (GFAP) staining] were neither hyperplastic nor hypertrophic. They were confined to the area of the original labelling site, and did not migrate either laterally across the pial margin or ventrally into the cortical layers. Knife wounding at the time of label application, either in the region of the label or distant from it, produced reactive astrocytes that were hypertrophic. These cells also did not migrate from the label site. These results suggest that astrocytes labelled by this method do not migrate in the absence of some transplant-derived stimulus even when stimulated by local wounding. PMID- 8244529 TI - Lines of glial precursor cells immortalised with a temperature-sensitive oncogene give rise to astrocytes and oligodendrocytes following transplantation into demyelinated lesions in the central nervous system. AB - Immortalised lines of murine glial precursor cells expressing the neomycin resistance gene and a temperature-sensitive mutation of the SV 40 T oncogene were established from cultures of oligodendrocytes and precursor cells infected with a replication-incompetent, helper-free retrovirus. At the permissive temperature (33 degrees C), they could be continually propagated in vitro and cells were present expressing the 04 antigen specific for glial precursor cells and oligodendrocytes. At 38 degrees C, where the expression of the T antigen is down regulated, cell division largely ceased. During early passage in vitro, limited differentiation to a more mature phenotype, as evidenced by expression of GFAP and the oligodendrocyte marker 01 was observed at both 33 degrees C and 38 degrees C. When transplanted into demyelinating lesions in the spinal cords of adult rats early passages of the lines yielded myelin-forming oligodendrocytes and astrocytes. Cells from later passages of the lines although failing to synthesise myelin still associated specifically with the demyelinated axons. These experiments demonstrate the retention of physiological properties of these oncogene-carrying glial cells when transplanted in vivo and suggest that such immortalised populations can be used for the isolation of molecules regulating glial cell function. PMID- 8244530 TI - Interleukin-6 production by brain tissue and cultured astrocytes infected with Theiler's murine encephalomyelitis virus. AB - Theiler's murine encephalomyelitis virus (TMEV) is known to interact with cells of the central nervous system (CNS). Here we report that, interestingly, it is a potent inductor of interleukin-6 (IL-6) in the CNS of infected animals and in pure cultures of astrocytes. Maximal IL-6 gene transcription in glial cells, as detected by bioassay and ELISA, was observed at 6 and 24 h after infection. Astrocytes from both SJL/J and Balb/c (strains of mice susceptible and resistant, respectively, to TMEV-induced demyelination) produced similar amounts of IL-6, measured in tissue culture supernatants. These results indicate that although an immunomodulatory effect can be exercised by IL-6 synthesized by astrocytes, it does not play a crucial role in immune-mediated demyelination induced by TMEV. PMID- 8244531 TI - Expression of janusin (J1-160/180) in the retina and optic nerve of the developing and adult mouse. AB - We have analyzed the expression of the oligodendrocyte-derived extra-cellular matrix molecule janusin (previously termed J1-160/180) in the retina and optic nerve of developing and adult mice using indirect light and electron microscopic immunocytochemistry, immunoblot analysis, and enzyme-linked immunosorbent assay. In the optic nerve, janusin is not detectable in neonatal and only weakly detectable in 7-day-old animals. Expression is at a peak in 2- or 3-week-old animals and subsequently decreases with increasing age. In the retina, expression increases until the third postnatal week and then remains at a constant level. In immunocytochemical investigations at the light microscopic level, janusin was found in the myelinated regions of the nerve with spots of increased immunoreactivity possibly corresponding to an accumulation of the molecule at the nodes of Ranvier. At the electron microscopic level, contact sites between unmyelinated axons, between axons and glial cells, and between axons and processes of myelinating oligodendrocytes were immunoreactive. Cell surfaces of astrocytes at the periphery of the nerve and forming the glial-limiting membrane, in contrast, were only weakly immunopositive or negative. In cell cultures of young postnatal mouse or rat optic nerves, oligodendrocytes and type-2 astrocytes, but not type-1 astrocytes were stained by janusin antibodies. In the oligodendrocyte-free retina, janusin was detectable in association with neuronal cell surfaces, but not with cell surfaces of Muller cells or retinal astrocytes. Our observations indicate that expression of janusin in the optic nerve and in the retina is developmentally differentially regulated and that other cell types, in addition to oligodendrocytes, express the molecule. Since the time course of janusin expression in the optic nerve coincides with the appearance of oligodendrocytes and myelin and since janusin is associated with cell surfaces of oligodendrocytes and outer aspects of myelin sheaths and is concentrated at nodes of Ranvier, we suggest that janusin is functionally involved in the process of myelination. PMID- 8244532 TI - Thyroid hormone up-regulates thyroid hormone receptor beta gene expression in rat cerebral hemisphere astrocyte cultures. AB - Oligonucleotide probes complementary to specific regions of three thyroid receptor cDNAs were used to study the effects of thyroid hormone on the expression of the mRNAs encoding two alpha (alpha 1 and alpha 2) and one beta thyroid (beta 1) receptors isoforms in rat cerebral hemisphere astrocyte cultures. Both genes are expressed by type 1 astrocytes. The levels of the alpha 1-, alpha 2-, and beta 1-mRNAs did not significantly change between day 8 and day 22, in cultures grown in the absence of thyroid hormone. L-triiodothyronine (L T3) treatment of the cultures increased the levels of beta 1-mRNAs by fivefold without changing either the levels of the alpha 1- and alpha 2-mRNAs or L-T3 binding capacity. The effect of L-T3 on beta 1-mRNAs was observed after 4 h of treatment and was independent of protein synthesis, suggesting that this effect is likely to be a direct one. Treatment of the cultures by cytosine arabinosine, a drug that kills dividing cells, specifically decreased level of the alpha 1- and alpha 2-mRNAs by 60% and 38%, respectively. Finally, by immunocytochemistry, we showed that the beta 1 receptor-immunoreactivity was either located in the perinuclear region and the cytoplasm or in the nuclei of astrocytes. Taken together with previous data obtained in neuronal cultures where no effect of L-T3 was observed on the levels of the beta 1-mRNAs, our findings indicate that the beta 1 gene is differentially regulated in neurons and astrocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244533 TI - Differential migration of astrocytes grafted into the developing rat brain. AB - Fetal and neonatal astrocytes migrate in specific patterns when transplanted into the adult rat host brain. However, it is unclear whether these astrocytes demonstrate the same degree of mobility during early brain development. In the present study, neonatal cortical, hippocampal, and hypothalamic astrocytes were collected from the brains of 1- to 3-day-old rats and placed in tissue culture. After 14 to 21 days, cultures enriched in astrocytes were harvested and labelled with either the fluorescent dye Fast Blue or fluorescein-labelled latex beads. They were then transplanted into the right frontal cerebrum of neonatal rats at 2, 5, 8, and 11 days postpartum. Seven days after transplantation, animals were sacrificed and their brains were fixed by immersion in aldehydes, sectioned on a cryostat, and examined with fluorescence microscopy. Transplanted astrocytes migrated along the corpus callosum, internal capsule, glial limitans, ventricular linings, and hippocampal structure. Labelled cells were also found in the contralateral hemisphere in day 2 brains. Migration in a radial fashion from the injection site toward the periphery was a particularly obvious pattern, and was most pronounced in these younger hosts. In days 5 and 8 rat brains, astrocyte migration became more restricted to the hemisphere of implantation. In 11-day-old host brains, hemispheric restriction and other region-specific influences became manifest and specifically modulated migration. Radial migration was absent in the 11-day-old host group except for cells of cortical origin. The observed results demonstrate that neonatal cortical, hippocampal, and hypothalamic astrocytes transplanted into the neonatal cerebrum migrate in patterns that are more extensive than in the adult brain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244534 TI - Effects of tumor necrosis factor on inward potassium current and cell morphology in cultured human oligodendrocytes. AB - The effects of recombinant human tumor necrosis factor-alpha (rhTNF-alpha) on inward rectifier potassium [K(IR)] currents and on cell morphology have been studied in cultured human oligodendrocytes. Cell-attached patches were used to isolate and record unitary currents through an inward rectifier K+ channel with a conductance of 23 pS. In control oligodendrocytes the mean open times showed an exponential dependence on patch potential with an e-fold decrease over a patch hyperpolarization of 28 mV. Treatment of oligodendrocytes with rhTNF (at 250 ng/ml for 24-48 h) had significant actions to diminish the mean open times of K(IR) relative to control values. At cell resting potential the mean open times were reduced by 60% after rhTNF application; the amplitudes of unitary currents or the extrapolated zero-current potentials were not significantly changed by the cytokine. The rhTNF treatments were not cytotoxic to cultured human oligodendrocytes; however, in some experiments rhTNF caused evident retraction of cell processes. PMID- 8244535 TI - A Ca(2+)- and pH-dependent K+ channel of rat C6 glioma cells and its possible role in acidosis-induced cell swelling. AB - The aim of the present study was to explore whether a change in membrane K+ conductance contributes to acidosis-induced swelling of cultured rat C6 glioma cells. Electrophysiological studies were performed using whole-cell and single channel recordings in combination with cell volume measurements in cell suspension by flow cytometry. Whole-cell recordings revealed a voltage-dependent K+ conductance. The predominant K+ channel in single-channel recordings with symmetrical high K+ concentrations was inwardly rectifying and had conductances of 35 and 15 pS, respectively. A raised internal free Ca2+ concentration and membrane depolarization increased the open probability of this channel. Internal acidosis (pH 6.4-5.4), on the other hand, reduced open probability and single channel conductance. Both whole-cell and single-channel K+ currents were blocked by quinidine (0.1-1 mM), which was therefore used to analyze the functional consequences of an inhibition of this conductance for cell volume. Thereby, quinidine (1 mM) produced a small (5%) and transient cell swelling of C6 glioma cells. In contrast, acidosis (pH 5.6) caused a much larger (about 20%) and maintained swelling. Since quinidine produced only a minor swelling of C6 cells, it is unlikely that inhibition of the K+ conductance caused acidosis-induced cell swelling. Other mechanisms, such as activation of ion transporters, must therefore be responsible. PMID- 8244536 TI - Endogenous opioid systems and the growth of oligodendrocyte progenitors: paradoxical increases in oligodendrogenesis as an indirect mechanism of opioid action. AB - Endogenous opioids inhibit nervous system development by inhibiting the proliferation of certain neuronal and glial progenitors. To determine whether opioids affect the growth of preoligodendrocytes, the effects of the endogenous opioid [Met5]-enkephalin were examined in preoligodendrocytes in primary mixed glial and preoligodendrocyte-enriched (> 98% pure) cultures. Proliferating preoligodendrocytes in mixed-glial or preoligodendrocyte-enriched cultures were continuously treated for a total of 40 h with either basal growth media (controls), 1 microM [Met5]-enkephalin, 1 microM [Met5]-enkephalin plus the opioid antagonist naloxone (3 microM), or naloxone alone (3 microM), and incubated in [3H]-thymidine (0.2 microCi/ml/4-6 h) after 34-36 h of opioid exposure. Opioid-dependent changes in DNA synthesis were assessed autoradiographically in O4-immunoreactive oligodendrocyte progenitors. Naloxone alone significantly decreased the rate of DNA synthesis and number of O4 immunoreactive preoligodendrocytes in mixed-glial cultures. However, naloxone and/or [Met5]-enkephalin did not affect DNA synthesis or the number of O4 immunoreactive preoligodendrocytes in cultures enriched in preoligodendrocytes. The results suggest that astrocytes, or perhaps another cell type, play a permissive role in opioid-dependent alterations in preoligodendrocyte proliferation. Endogenous opioids affect the genesis of neural cells by both direct and indirect mechanisms. PMID- 8244537 TI - Glial calcium. AB - This review summarizes current knowledge relating intracellular calcium and glial function. During steady state, glia maintain a low cytosolic calcium level by pumping calcium into intracellular stores and by extruding calcium across the plasma membrane. Glial Ca2+ increases in response to a variety of physiological stimuli. Some stimuli open membrane calcium channels, others release calcium from intracellular stores, and some do both. The temporal and spatial complexity of glial cytosolic calcium changes suggest that these responses may form the basis of an intracellular or intercellular signaling system. Cytosolic calcium rises effect changes in glial structure and function through protein kinases, phospholipases, and direct interaction with lipid and protein constituents. Ultimately, calcium signaling influence glial gene expression, development, metabolism, and regulation of the extracellular milieu. Disturbances in glial calcium homeostasis may have a role in certain pathological conditions. The discovery of complex calcium-based glial signaling systems, capable of sensing and influencing neural activity, suggest a more integrated neuro-glial model of information processing in the central nervous system. PMID- 8244538 TI - Blood transfusion and injured patients. PMID- 8244539 TI - Use of blood transfusions by helicopter emergency medical services: is it safe? AB - All injured patients given prehospital blood transfusions by flight nurses of the 'Lifeflight' Helicopter Emergency Medical Service (HEMS) programme based at Emanuel Hospital, Portland, Oregon, USA, between 1985 and 1991 were included in the study. Of the 112 patients, there were no documented adverse reactions directly related to the administration of O-negative packed red cells, although one patient suffered transient shortness of breath. It is a safe and practical prehospital procedure. Further studies are required to assess the advantages over other fluids. PMID- 8244540 TI - Caveat surgeon: do orthopaedic surgeons take adequate precautions against blood borne viral infections, in particular the human immunodeficiency virus (HIV)? AB - One year ago the British Orthopaedic Association issued guidelines for the prevention of cross-infection with special reference to HIV and the hepatitis viruses. We were interested to establish whether the guidelines were being widely applied and whether they had changed general orthopaedic practice. We distributed a questionnaire to Scottish Orthopaedic Fellows, Associates of the BOA, and orthopaedic trainees. With a 70 per cent return rate, it would appear that the recommendations are not adhered to in full. Of respondents, 84 per cent were immunized or undergoing immunization against hepatitis B. In all, 30 per cent were operating on high-risk patients on a monthly basis, 60 per cent thought that their current practice was low risk, and only 15 per cent thought that their future practice would be high risk; 81 per cent were concerned and yet only 60 per cent had altered their practice. It is of some concern that orthopaedic surgeons may not take the threat of HIV cross-infection seriously enough and do not consider precautions mandatory. Further pressure and support from the BOA may be necessary to encourage a change in orthopaedic practice as the threat of HIV is increasing. PMID- 8244541 TI - Resolving the casualty evacuation conflict. AB - One of the most contentious issues surrounding the prehospital management of the injured is the decision to stabilize on site or to rapidly evacuate the patient. Although there are vociferous proponents for both views, there appears to be little thought given to the rationalization of these decisions, and the subject has only merited lip-service in previous texts. Treatment choices and decisions in the prehospital environment are notoriously difficult to make. The doctor, unlike ambulance paramedics, does not necessarily work to didactic protocols and, if suitably trained, is at liberty to commence advanced resuscitation procedures at the roadside. However, these may be time consuming and, to be of greatest benefit to the patient, a fine balance between resuscitation and evacuation must be drawn. This paper aims to discuss some of the key issues which need to be considered in the prehospital scenario, such as treatment triage, evacuation priorities, mode of transportation and hospital destination. The end result must be to eliminate the medical 'bottleneck' and to afford early and appropriate definitive care to those in most need. PMID- 8244542 TI - Post Gulf war explosive injuries in liberated Kuwait. AB - We collected the statistics of the victims affected by explosive injuries in liberated Kuwait during the period March to December 1991. Included were 1679 patients treated at nine different hospitals in Kuwait. As members of a trauma team, we, the plastic surgeons, were involved in the care of 152 victims of explosive injuries admitted at Al-Razi Orthopaedic Hospital. The records of these patients were reviewed in detail and the results were analysed. Of the patients, 58 had penetrating wounds and 94 explosive blast wounds. In all, 69 patients (45.3 per cent) were found to have associated fractures, 8 (5.3 per cent) had vascular injuries and in 10 (6.5 per cent) a nerve was also involved. There were 41 patients (26.9 per cent) who required amputations. The mainstay of surgical treatment was adequate wound excision, skeletal fixation and soft tissue coverage. Split-skin grafts were used in 34 patients, local flaps in seven and free flaps in four. Most patients healed within 2-3 weeks. There were 10 patients with explosive blast wounds who developed infection and needed staged wound excision before providing final coverage. PMID- 8244543 TI - A cross-matching policy for fractures of the proximal third of the femur. AB - Cross-matching and transfusion policies for patients with a fracture of the proximal third of the femur are poorly formulated. A series of 90 patients who underwent operation for such fractures at a district general hospital were evaluated. Unnecessary cross-matching was most frequent in intracapsular fractures, and discrepancies in indications for postoperative transfusions were demonstrated. If postoperative transfusion is indicated for haemoglobin levels of less than 9 g/dl, we recommend routine preoperative cross-matching only for patients admitted with trochanteric fractures and haemoglobin levels between 10 and 12 g/dl. This practice is safe and is a potential area for savings in cost and resources. PMID- 8244544 TI - A finite element analysis of the effect of pin distribution on the rigidity of a unilateral external fixation system. AB - The effect of pin distribution on the rigidity of a unilateral external fixation system has been investigated using finite element techniques. The results show that spreading the pins out along the entire length of the bone is more stable than grouping the pins in clusters as is customary. PMID- 8244545 TI - A comparative study of early motion and immediate plaster splintage after internal fixation of unstable fractures of the ankle. AB - We reviewed 47 patients following operatively treated ankle fracture-dislocation, at an average of 15 months after injury, to assess the outcome of two different postoperative regimens. Of the 47 patients, 27 received early active and passive ankle exercises, and 20 patients received immediate plaster splintage. Patients were assessed clinically by an independent surgeon and subjective, objective and radiological criteria recorded. No significant difference was apparent between the two groups on any of the criteria, although the early movement group contained more patients who were completely pain free, had a normal gait and no radiological signs of arthrosis (P < 0.05). This was achieved at the expense of a longer stay in hospital (average 10.2 days versus 7.4 days for plaster splintage) and more ankle swelling. PMID- 8244546 TI - Muscle function after plated and nailed femoral shaft fractures. AB - We studied retrospectively 14 patients who had been nailed and 12 patients who had been plated for femoral shaft fractures respectively 37 and 67 months previously. Isokinetic thigh muscle function measurements showed quadriceps strength to be fully restored in nailed patients, while there was a moderate persisting median deficit among plated patients. There was a similar moderate deficit in flexion strength for both groups. Strength restoration did not correlate with age at fracture or fracture level in the femur. It did, however, correlate strongly with the interval between fracture and study for flexion strength among nailed patients. It may be that this parameter also is eventually normalized in nailed patients. PMID- 8244547 TI - External fixator pin placement in the forearm: how safe is it? An anatomical study. AB - An anatomical study was undertaken using cadaveric forearms to identify the best technique and angle of pin placement for external fixator pins. We have found that inserting pins through stab incisions risks transfixion of the underlying structures and leads to an increased morbidity. We conclude that inserting pins under direct vision is much safer and that the optimum angle of pin insertion is in the coronal plane, parallel to the plane of the metacarpals. PMID- 8244548 TI - Use of Autosuture Surgiport for pleural drain insertion. AB - The insertion of pleural drains (tube thoracostomy) is associated with serious complications in up to 10 per cent of cases. A safe and efficient technique of tube thoracostomy using the Autosuture Surgiport is described. PMID- 8244549 TI - Injuries to the heart. AB - During a 12-year period, 36 patients with cardiac injuries were recorded. Nine suffered penetrating injuries, nine blunt ruptures and 18 contusions. Only two patients with blunt rupture survived, in contrast to six out of seven patients with stab wounds to the heart. This reflects that heart stabs are easier to diagnose than blunt ruptures, and that the associated injuries are the most important factor for survival in patients with blunt injuries. All clinical manifestations of the heart injuries were present within the first 12 h. Cardiac monitoring of haemodynamically stable patients beyond this period may not be necessary. PMID- 8244550 TI - Whiplash injury and surgically treated cervical disc disease. AB - The incidence of a previous whiplash injury in 215 unselected patients who underwent an anterior cervical discectomy and fusion between 1988 and 1991 was found to be twice that of a control population of 800 general orthopaedic outpatients (P < 0.05). The mean age at which the whiplash injury occurred in the surgical group was 37 years and in the control group 36 years. The mean age at operation of those patients with a previous whiplash injury (45 +/- 12 years) was significantly less than those patients without a previous whiplash injury (55 +/- 14 years, P < 0.001). This study provides further evidence that whiplash injury causes structural changes predisposing to premature degenerative disc disease. PMID- 8244551 TI - Free fibular graft still has a place in the treatment of bone defects. AB - We report on 27 patients illustrating the use of non-vascularized single fibular strut graft, augmented with a corticocancellous bone graft to bridge bone defects. The indications were varied and included infection, fracture with bone loss, non-union, bone tumour, bone cyst and congenital pseudarthrosis. Primary union was achieved in 92 per cent. Stress fracture occurred in 26 per cent and no significant fibular graft hypertrophy occurred. The aim of this paper is to show that the non-vascularized single fibular graft, if augmented with corticocancellous bone graft along its whole length, is a simple procedure that is still valid to bridge bone defects. PMID- 8244552 TI - Vessel-like structures in the callus cartilage. AB - The recent claim that the callus cartilage contained vessel-like structures has been investigated by new immunocytochemical methods. The fracture callus was exposed to monoclonal antibodies which recognized damaged or regenerating endothelial cells. The proteolytic enzyme urokinase which is expressed by migrating endothelium was also studied. No vascular structures were identified by these methods which suggests that the callus cartilage is truly avascular. PMID- 8244553 TI - Patterns of meniscal tears associated with anterior cruciate ligament lesions in athletes. AB - In this study, 400 clinically anterior cruciate ligament (ACL) deficient knees were arthroscoped and studied prospectively in the period January 1986 to April 1992. An ACL tear was always confirmed, and 41 per cent of these patients did not have an associated meniscal tear. In 30.25 per cent the lateral meniscus was torn; in 21.25 per cent the ACL tear was associated with a medial meniscus tear, and in the remaining 7 per cent both menisci were torn. The most frequently associated meniscal injury was the bucket handle tear of the medial meniscus (9 per cent), followed by the posterior horn tear of the lateral meniscus, which showed the same frequency as the ragged (or degenerated) tear of the lateral meniscus (6 per cent). The horizontal tear of the posterior part of the lateral meniscus showed a prevalence of 4.3 per cent. This picture is probably dependent on a secondary referral nature of the centre surveyed, in which the average time between injury and arthroscopy was 23.3 months. PMID- 8244554 TI - Hazard of ionizing radiation to trauma surgeons: reducing the risk. AB - A study aimed to establish the level of radiation exposure to orthopaedic surgeons involved in the care of injured patients; parts of the body most at risk, and to establishing whether surgeon control of X-ray image intensification reduced the risk. This was conducted on five orthopaedic surgeons regularly involved in musculoskeletal care. Radiation dosage (in millisieverts (mSv) to the body, neck, eyes and hands, was measured by means of dosimeters applied to these areas, before and after surgeon-controlled use of X-ray image intensification. Although all doses measured were within current safety guidelines (1.25 mSv total body dose/month, 3.75 mSv eye dose/month and 12.5 mSv extremity dose/month), the hands were most at risk (maximum recorded dosage 3.95 mSv/month). Control by the surgeons of X-ray image intensification significantly reduced exposure of the hands (P < 0.05). PMID- 8244555 TI - Gradual closure of fasciotomy wounds using a vessel loop shoelace. PMID- 8244556 TI - Sparing of deep rectal sensation after spinal cord injury. PMID- 8244557 TI - An unusual proximal tibial epiphyseal injury. PMID- 8244558 TI - Acute onset meralgia paraesthetica after fracture of the anterior superior iliac spine. PMID- 8244559 TI - Internal fixation of a fracture of the distal femur with ipsilateral osteoarthritis of the hip. PMID- 8244560 TI - Management of skeletal trauma in the United Kingdom. PMID- 8244561 TI - Butler's life review: how universal is it? AB - In the nearly thirty years since Butler first proposed the life review, the notion that older persons review their lives in the face of approaching death has become firmly entrenched in both the literature and practice of gerontology. This article examines the commonly-held assumptions underlying the life review and reviews what previous studies have found in testing these assumptions. Findings from a recent study of centenarians are also presented that further call into question some aspects of the life review. PMID- 8244562 TI - Themes in reminiscence groups with older women. AB - The preponderance of older women in the United States is apparent to all professionals in health care for elderly individuals. Because there is much emphasis on health care promotion for all age groups, providers of health care seek strategies for health promotion for older women. Although the literature is replete with information on reminiscence in general, and specifically reminiscence group therapy, there has been little discussion about the use of themes in reminiscence therapy groups. There are no publications about themes appropriate for reminiscence groups composed solely of older women. This article is based on three research studies and analyzes themes used in the three protocols for those studies. Appropriate themes for an all women's group are discussed and implications for practice are suggested. PMID- 8244563 TI - Subjective age and health perceptions of older persons: maintaining the youthful bias in sickness and in health. AB - Self-reports of 250 persons fifty years of age and older confirm the increasing bias toward reporting a more youthful age as one increases in years. Optimistic perceptions of health care also maintained in older persons. Results from two subsets of this sample (N = 48) further indicate that the youthful and optimistic bias occurs in older persons with poorer and failing health (N = 23) as well as for persons in stable and good health (N = 25). Given the importance of self perceptions in quality of life and in determining survivability, and given the indication that such measures are modifiable, it is suggested that future research be aimed at identifying those self-perceptions of health and age that are most susceptible to intervention. PMID- 8244564 TI - Caregivers' reactions to the physical appearance of a person in the final stage of dementia as measured by semantic differentials. AB - The semantic differential (SeD) technique was applied to 158 caregivers from a nursing home in the northern part of Sweden. The questionnaire contained fifty eight bipolar scales of adjective pairs and the interviewees indicated their reactions to a described picture of a severely demented person: A factor analysis revealed three dimensions; an ethical one, an esthetical one, and one about the person's own feelings. The fifty-eight scales were mostly rated toward the negative poles. The severely demented person was rated as painful, apathetic, suffering, weak, afraid, sad, cold, dark, rough, and ugly. Four years later a comparable group of caregivers (n = 93) answered a revised questionnaire containing the 26 SeD scales with factor loadings > .50 for the picture of the severely demented person. The result was nearly identical and alternative interpretations are discussed. PMID- 8244565 TI - Age and typicality judgments of stereotypes of the elderly: perceptions of elderly vs. young adults. AB - Healthy community-dwelling elderly individuals assessed the typicality and approximate age of individuals representing ten stereotypes of elderly people identified in prior research with young adults. Results showed that, as predicted, the elderly adults saw the stereotypes as less typical of elderly individuals than did the young adults, although both groups agreed on which stereotypes were most typical of the general elderly population. Analysis of age judgments revealed that the elderly respondents, in comparison to the young adults, were more likely to use the old-old age ranges to describe the stereotypes, and much less likely to associate the positive stereotypes with the young-old age ranges. Both elderly and young adults tended to associate negative stereotypes with the older elderly age ranges. PMID- 8244566 TI - Race, gender, and the effect of social supports on the use of health services by elderly individuals. AB - This study examines the effects of social supports on the use of health services by elderly individuals, and whether those effects differ by race or gender. Multiple regression analysis of the 5,151 baseline Longitudinal Study on Aging (LSOA) respondents revealed that social supports, mainly church attendance, increased the R2s when estimated separately within race and gender groups in a statistically significant manner. The magnitude of these increases, however, was small suggesting that their unique contribution may be minimal. The effects of the social support variables on the use of health services differed by race and gender. These differences, however, were not consistent across the various measures of health services utilization. PMID- 8244567 TI - Working group report on hepatitis viruses. PMID- 8244568 TI - Specific expression of the pancreatic-secretory-trypsin-inhibitor (PSTI) gene in hepatocellular carcinoma. AB - Twenty hepatocellular carcinomas (HCC) were analyzed by Northern blotting to test the expression of pancreatic secretory trypsin inhibitor (PSTI). This gene was expressed in all HCCs, but not in other tumors, including mammary, thyroid, pulmonary and ovarian cancers. Some gastric and colonic cancers weakly expressed PSTI. Among cell lines examined in a similar manner, PSTI was expressed in all of 4 derived from hepatoma. On the other hand, among 15 cell lines derived from cancers other than hepatoma, only 3, derived from pancreatic, colonic and gastric cancers, weakly expressed PSTI. A CAT assay using a deletion set of the 5' region from the cloned PSTI gene has shown that in hepatoma cell lines, the expression of this gene is dependent on the presence of 2 regulatory regions that include an IL-6 responsive elements and an AP-I-binding site. However, in non-hepatoma cell lines, the 2 regulatory regions are not necessary for expression. The blood level of PSTI in 27 patients with HCC was significantly increased, and it was positively correlated with tumor size, suggesting that specific expression of PSTI in HCC causes this effect and that elevated blood level of PSTI without inflammation indicates the presence of HCC. PMID- 8244569 TI - Intratumoral microvessel density and p53 protein: correlation with metastasis in head-and-neck squamous-cell carcinoma. AB - Squamous-cell carcinoma of the head and neck includes a heterogeneous group of tumours of the upper air and food passages for which prognosis is difficult to assess. In fact, patients in comparable stages may have diverse clinical courses and responses to similar treatments. In order to better define the prognosis of each patient there is therefore a need to identify novel biological markers which reflect more accurately growth rate, progression and metastatic potential of each tumour. We assessed whether metastases correlate with microvessel counts (i.e. intratumoral vascularity) using the CD-31 monoclonal antibody (MAb) and p53 mutant protein expression, determined in the primary by immunocytochemical methods in 70 patients with locally advanced head and neck cancer. Patients were treated with concurrent chemo-radiotherapy; 50 of these presented loco-regional node metastasis at diagnosis whereas 3 cases, initially node-negative, developed distant metastasis during the period of observation. No feature was predictive for objective response to treatment. The overall mean and median blood vessel density at "hot spots" was 37.42 and 36, respectively, and 57% of the tumours expressed p53 mutant proteins. These 2 biological markers were significantly associated. Patients with metastases (loco-regional and distant) had a significantly higher mean blood-vessel density than those without tumour spread. Also, patients with p53-positive (+/++) tumours had a significantly higher incidence of metastasis than those with negative ones. Multivariate analysis showed that both vascularity and stage, but not p53 expression, are significant and independent predictors of metastasis in this series. PMID- 8244570 TI - Parity and hepatocellular carcinoma. A population-based study in Sweden. AB - The association between parity and hepatocellular carcinoma (HCC) was studied using a data-base generated by linking 2 Swedish nation-wide registries; the Cancer Registry and the Fertility Registry. Among women born between 1925 and 1960, 133 patients with HCC recorded in the Cancer Registry between 1958 and 1984 were compared with 665 age-matched controls. In this nested case-control study there was no positive association between parity, age at first birth or frequency of twinning on the one hand, and risk of HCC on the other. It appears that the positive association between parity and HCC previously reported is limited to cases of HCC caused by chronic infection with hepatitis B virus; these cases represent only a small fraction of HCC cases in Sweden. PMID- 8244571 TI - Epstein-Barr virus subtype distribution in angioimmunoblastic lymphadenopathy. AB - The tissues of 16 patients bearing a T-cell lymphoma of angioimmunoblastic lymphadenopathy type (AILD-TCL) were investigated for the distribution of Epstein Barr virus (EBV) subtypes 1 and 2. EBV-association had been proven in these cases by polymerase chain reaction (PCR) for EBV-DNA, in situ hybridization (ISH) for EBV-encoded small nuclear RNAs (EBER) and immunohistology for EBV-encoded latent membrane protein (LMP). PCR and EBER-ISH produced mostly identical results, but some cases were positive with only one of the 2 methods employed. LMP was detected in a few large cells of 8/13 cases. Twelve cases were investigated for the distribution of EBV subtypes. One case contained EBV genome of subtype 2, 3 cases contained subtype 1 and 4 cases contained both subtypes. Four cases could not be typed. These findings suggest that in AILD, as in AIDS-associated lymphomas and lymphomas of the lethal midline granuloma type, subtype 2 of EBV may occur, perhaps in relation to an immunodysfunction developing progressively in these patients. PMID- 8244572 TI - Expression of alpha B-crystallin in human brain tumors. AB - We have previously shown that alpha B-crystallin is a heat-shock protein which specifically accumulates in response to the expression of c-Ha-ras and v-mos oncogenes in mouse NIH 3T3 fibroblasts. Elevated levels of alpha B-crystallin mRNA or protein were shown to be associated with pathological conditions of the brain. Therefore, we have examined the expression of alpha B-crystallin in normal human brains and brain tumors by Western blot analysis. alpha B-crystallin is moderately expressed in adult but not fetal brain. Elevated levels of alpha B crystallin expression are observed in glial tumors such as astrocytoma, glioblastoma multiforme, and oligodendroglioma. alpha B-crystallin in these tumors is predominately unphosphorylated. High amounts of accumulated alpha B crystallin in astrocytic tumors are preferentially found in the more aggressive stages. Glioblastoma multiforme is exceptional in that high alpha B-crystallin expression is observed in only one half of the analyzed samples whereas no alpha B-crystallin could be detected in the other. These results indicate that alpha B crystallin may be a useful biochemical marker for studying the pathogenesis of various human brain tumors. PMID- 8244573 TI - An analysis of a melanoma epidemic. AB - Between 1986 and 1988 the annual incidence of invasive melanoma in the Hunter area of New South Wales, Australia, almost doubled to 52.5 per 100,000 in men and 42.9 per 100,000 in women. These rates have been maintained and are similar to those reported for 1987 in Queensland, Australia, which are the highest in the world. Most of the increase in incidence was in melanomas less than 1.50 mm in thickness, and adults of both sexes and all ages were affected. Thicker melanomas also increased in incidence but only in adults 45 years and older, and mainly in men. An analysis of health insurance data on treatment of skin lesions and data from a histopathology laboratory suggested that diagnosis and treatment of skin lesions generally in the Hunter area had increased almost 2-fold over this period. Advancement of the time of diagnosis and a real increase in incidence were likely explanations for some of the observed trends. Increasing diagnosis of a non-metastasising form of thin melanoma, consequent upon increasing removal of pigmented skin lesions by medical practitioners, may also explain some of the observed increase in the incidence of the disease. This possibility has important implications for proposed population screening programs, and methods are needed to distinguish such lesions, if they exist, from potentially fatal melanoma. PMID- 8244574 TI - Stromelysin-3 expression in breast cancer biopsies: clinico-pathological correlations. AB - Stromelysin-3 expression was studied by Northern blotting in 222 tissue samples including primary and metastatic breast carcinoma and normal breast tissue. Uninvolved breast tissue from mastectomy specimens, normal breast tissue from reduction mammoplasties and normal lymph nodes did not contain stromelysin-3 mRNA. About 62% of primary and metastatic breast carcinomas, but only 1 of 10 in situ ductal carcinomas, expressed stromelysin-3. Stromelysin-3 mRNA was found more often in estrogen-receptor-positive carcinomas and in histological grade-1 carcinomas. There was no significant correlation between stromelysin-3 expression and other prognostic factors, including tumor size, lymph-node involvement, age of patient, vascular invasion and cathepsin-D. PMID- 8244575 TI - Serologic response in human papillomavirus-associated invasive cervical cancer. AB - Human papillomavirus (HPV) transforming proteins E6 and E7 are uniformly expressed in HPV-associated cervical cancer. Our objective was to measure antibodies to HPV-16 E6 and E7 proteins in cervical cancer patients using an assay which would detect antibodies to conformational epitopes. Serum specimens obtained from two case-control studies of HPVs and cervical cancer were tested. The studies were performed in Cali, Colombia, South America and in 9 provinces of Spain. Cases consisted of women with invasive cervical cancer associated with HPV 16 or other HPV types and women with HPV-16-associated high-grade cervical intra epithelial neoplasia (CIN-3). Controls for invasive cases and CIN-3 cases were women who had no cytologic abnormalities and who were matched for age and country of residence. Serum antibodies to HPV-16 E6 and E7 proteins were detected by radio-immunoprecipitation of in vitro translated proteins. Antibodies to the E6 and E7 protein were observed among 56% and 43%, respectively, of invasive cases and 1.7% and 4.1%, respectively, of controls. Antibodies to either protein were detected in 72% of sera from invasive cases and 5.8% of sera from controls. High antibody reactivity and antibodies to both proteins were found almost exclusively in invasive cases. The frequency of antibodies to the E6 protein and the E7 protein among CIN-3 cases did not differ significantly from the CIN-3 controls. Five women with HPV-18-associated invasive cervical cancer were negative for serum antibody to HPV-16 E6 and E7 proteins. Antibodies to HPV-16 E6 and E7 proteins appear to be partially virus-specific and disease state-specific markers of HPV-associated cervical cancer. PMID- 8244576 TI - The predominant mRNA class in HPV16-infected genital neoplasias does not encode the E6 or the E7 protein. AB - Human papillomavirus (HPV) type 16 is strongly implicated in the development of progressive neoplasias of the uterine cervix. Its oncogenic potential is decisively determined by the activity of the early gene products E6 and E7. To look for changes in the expression of these genes during tumour progression we cloned subgenomic fragments of HPV16 into RNA expression vectors, which allowed the generation of 35S-labelled riboprobes specific for distinct mRNA classes. Four constructs were made to differentiate between transcripts starting upstream of the E6 ORF or the E1 ORF, and one probe was specific for unspliced E6/E7 region transcripts. Five other constructs were used to identify transcripts covering the E1, E2, E4, L1 and L2 regions. With the help of these constructs, we analyzed by in situ hybridization 2 low-grade intraepithelial neoplasias of the vulva, 1 high-grade neoplasia of the cervix as well as 4 vulvar and 3 cervical carcinomas. Transcripts from the E1, E2, E4, L1 and L2 region that were consistently detected in the differentiated layers of benign lesions were variably expressed in precancers and carcinomas. None of the investigated cases revealed detectable amounts of unspliced E6/E7 transcripts with a coding potential for a full-length E6 protein. In benign lesions, the E7 transcripts were confined to isolated nuclei of differentiated cells, whereas high-grade lesions and invasive cancers showed elevated levels of equally distributed E7 specific signals in the cytoplasm of all tumour cells. The most abundant transcripts observed in intraepithelial neoplasias and in invasive cancers appear to initiate within ORF E7 and therefore have no coding potential for full-length E6 and E7 proteins. Our data show that the actual level of E7-specific transcripts in cancers is lower than anticipated from earlier studies using an ORF E6/E7-specific probe that hybridizes with the 5'-ends of the abundant mRNA class. PMID- 8244577 TI - Helicobacter pylori infection in Japanese patients with adenocarcinoma of the stomach. AB - To examine the association of Helicobacter pylori infection with adenocarcinoma of the stomach in Japanese patients, we studied 29 patients and 58 matched controls. Ascertainment of H. pylori status was based on the presence of specific IgG to H. pylori. For the entire group, an association of this infection with gastric adenocarcinoma was suggested but not statistically significant. For patients in a putatively high-risk subgroup (non-cardia tumors and age < or = 70 years), the association was significant. Assays detecting serum IgA to whole H. pylori cells and cytotoxin, IgG to cytotoxin and Hp54K (the heat-shock protein homolog) and serum neutralization of cytotoxin activity each showed clear differences between H. pylori-infected and uninfected persons in this population. However, for none of these assays was there a significant difference between values for H. pylori-infected persons with or without gastric cancer. Thus, while H. pylori infection was associated with non-cardia gastric cancer in Japanese persons < or = 70 years of age, use of these additional serologic markers did not define additional factors that might be associated with increased risk. PMID- 8244578 TI - Characterization of cell lines established from Merkel-cell ("small-cell") carcinoma of the skin. AB - Six cell lines have been established from different biopsies of the rare small cell skin cancer of neuro-endocrine origin known as Merkel-cell carcinoma (MCC). These were established from metastatic lesions and have now been in culture for periods varying from 1 to 3 years. All lines grow as suspension cultures and exhibit typical MCC cytological features of small round cells with little cytoplasm, round nuclei and tight junctions. There was variation in the number and size of nucleoli and dense-core granules. We present details of their immunohistochemistry and growth characteristics in culture. The latter were similar to small-cell lung-cancer (SCLC) lines and can be subdivided, by their appearance, into type I, type II and type III in the same way as SCLC lines. This classification may be of significance for the prognosis and management of MCC. PMID- 8244579 TI - Uncoordinate induction and differential regulation of HLA class-I and class-II expression by gamma-interferon in differentiating human neuroblastoma cells. AB - Recombinant gamma-interferon (IFN-gamma) has recently been shown to be one of the most effective inducers of neuroblastoma (NB) cell differentiation. Since increasing evidence indicates that expression of MHC class-I and class-II antigens by tumour cells is important for immunorecognition and cell targeting, we tested whether induction of NB cell differentiation by IFN-gamma is followed by expression of HLA class-I and class-II molecules. LAN-5 human NB cell line completely lacks HLA class-I antigens. Their expression was induced in a dose dependent manner by IFN-gamma. HLA class-II molecules are also absent on LAN-5 cells, but only DP antigens were dose-dependently induced by IFN-gamma, while DR and DQ molecules were unaffected by the treatment. To confirm and extend the immunological data to all the class-II molecules, we performed Northern blot analysis, observing that DP alpha mRNA was induced in a dose- and time-dependent manner. DO beta and DZ alpha genes were also induced peaking after 3 days of IFN gamma treatment. DR beta and DQ beta genes, which were not induced by IFN-gamma, gave a normal pattern of enzyme restriction by Southern blot. To get an insight into the regulation of HLA class-II gene expression in the neuronal model, we measured the decline of the steady-state HLA class-II mRNA. DO beta mRNA rapidly returned to baseline level after removing IFN-gamma, while the decay rates of DP alpha and DZ alpha mRNA were very slow. This might indicate different regulation at the post-transcriptional level for DO beta mRNA with respect to DP alpha and DZ alpha mRNA. To strengthen these findings we evaluated the half-lives of the mRNA after IFN-gamma induction by means of actinomycin D treatment. HLA-DO beta mRNA had a shorter half-life, while DZ alpha and DP alpha had a longer decay rate. Finally, we report that treatment of LAN-5 cells with cycloheximide did not alter the rate of transcription of the HLA-DP alpha gene, suggesting that no protein factor(s) is/are needed to maintain DP alpha gene expression. PMID- 8244580 TI - A CD16/CD30 bispecific monoclonal antibody induces lysis of Hodgkin's cells by unstimulated natural killer cells in vitro and in vivo. AB - In order to target NK cells against the Hodgkin's-derived cell line L540, we developed bispecific monoclonal antibodies (Bi-MAbs) by somatic hybridization of the 2 mouse hybridoma cell line HRS-3 and A9 which produce monoclonal antibodies (MAbs) with reactivity against the Hodgkin and Reed-Sternberg cell-associated CD30 antigen and the CD16 antigen (Fc gamma III receptor), respectively. The CD16 MAb-producing cell line A9 was selected as a partner for HRS-3 because of its efficiency in inducing lysis of the A9 hybridoma cells by resting NK cells. The hybrid hybridoma cell line HRS-3/A9 produced the supernatant with the strongest bispecific reactivity and was repeatedly subcloned and used for ascites production. Crude supernatant and purified HRS-3/A9 Bi-MAb triggered specific lysis of the CD30+ Hodgkin's-derived cell line L540, but not of the CD30- cell line HPB-ALL by unstimulated peripheral-blood lymphocytes and NK-cell-enriched populations. Moreover, treatment of SCID mice bearing heterotransplanted human Hodgkin's tumors with HRS-3/A9 and human peripheral blood lymphocytes induced specific complete tumor regression in 10/10 animals. We thus report successful tumor treatment in an in vivo model using NK-cell-associated Bi-MAbs and show that the Bi-MAb HRS-3/A9 is an efficient promoter of the anti-tumor effects of NK cells in vitro and in vivo. PMID- 8244581 TI - Quantitative and/or qualitative changes in the p21-H-ras post-translational products in regenerating liver and during hepatocarcinogenesis. AB - Using Western immunoblot analysis with the Ras-10 monoclonal antibody (MAb), we characterized 4 post-translational products of the c-H-ras gene in rat tissues, and showed that they were readily distinguished from the normal p21-N-ras or Val12 mutant p21-H-ras products. In the present study, we used this approach to compare the electrophoretic pattern of p21-H-ras during chemically induced hepatocarcinogenesis in rats with that in control liver samples. Three types of pattern were defined in the liver samples taken at various stages of malignant progression. The first type, like all the 18 normal samples analyzed, was characterized by a predominant intensity of spot a (corresponding to the palmitoylated p21-H-ras product). This was observed in all the samples at the stage of foci and only in a proportion of nodular or tumoral tissues. The second type of pattern deviates from this normal basal pattern by a higher relative level in one or more of the precursors of the fully processed p21-H-ras product. It was observed in all liver samples at the stage of nodules and in a proportion of tumors, but also in all samples from fetal or regenerative liver, thus suggesting an association with high proliferative activity of the hepatocytes. The third type of pattern was characterized by the presence of spots never detected in any of the normal rat tissue or cells that we investigated. Abnormal spots were observed in nodular liver samples and in hepatocarcinomas, indicating that they probably correspond to mutant p21-H-ras products. The fact that the 2 types of abnormal products were not constantly associated with neoplasms indicates that if they play a role in their induction or maintenance this may also be achieved in an independent way, perhaps, but not necessarily, involving another mutation of the ras gene. PMID- 8244582 TI - Decreasing the level of translation initiation factor 4E with antisense RNA causes reversal of ras-mediated transformation and tumorigenesis of cloned rat embryo fibroblasts. AB - Transformation of cloned rat embryo fibroblasts (CREF) with the T24-ras oncogene results in loss of contact inhibition, growth in soft agar and tumor formation in nude mice. Previously we showed that in such cells (CREF T24), the phosphorylation rate of protein synthesis initiation factor 4E (eIF-4E) is increased, correlating with an increase in the general rate of protein synthesis. In the present study, we have expressed antisense RNA complementary to eIF-4E mRNA in CREF T24 cells using a stably integrated vector. Cells expressing antisense RNA (CREF T24/AS) contained 30-50% of the normal level of eIF-4E and exhibited many of the properties of untransformed cells. CREF T24 had a spindle shaped, refractile appearance, whereas CREF T24/AS grew in ordered, parallel patterns and exhibited contact inhibition similar to untransformed CREF. The rates of growth and protein synthesis in CREF T24/AS were decreased compared to CREF T24 but were not as low as in CREF. The efficiency of growth in soft agar was 11-fold lower for CREF T24/AS compared with CREF T24. The latency period for tumor formation in nude mice was increased from 8 days for CREF T24 to 17-27 days for CREF T24/AS and various clonal lines derived from them. Cell lines established from these CREF T24/AS-derived tumors were shown to have partially regained the eIF-4E levels characteristic of CREF T24. These results demonstrate that many of the phenotypic alterations associated with ras-induced malignant transformation can be reversed by a moderate reduction of the translational initiation capacity and therefore may be mediated through a translational mechanism. PMID- 8244583 TI - Effect of buthionine sulfoximine on PtII and PtIV drug accumulation and the formation of glutathione conjugates in human ovarian-carcinoma cell lines. AB - Glutathione (GSH) has often been implicated in the mechanism of resistance to platinum anti-cancer drugs. It has been suggested that GSH may reduce the cytotoxicity of these drugs by forming inactive conjugates and by enhancing the repair of DNA-platinum crosslinks. In the present study we have examined the effect of D,L-buthionine-S,R-sulfoximine (BSO) pretreatment on the accumulation of platinum in a sensitive (CHI) and 2 relatively resistant (SKOV-3, HX/62) human ovarian-carcinoma cell lines following exposure to PtII- (cisplatin, carboplatin) and PtIV-drugs (tetraplatin). The metabolism of cisplatin and tetraplatin (particularly the extent of platinum-GSH conjugate formation) in the presence and absence of BSO pre-treatment was also examined in these cell lines. BSO pre treatment reduced the accumulation of PtII but not that of PtIV drugs in the relatively resistant SKOV-3 and HX/62 cell lines. It had no effect on the accumulation of either class of drugs in the sensitive CHI cells. Metabolism studies with cisplatin showed that the SKOV-3 and HX/62 cells, which contained 2- to 3-fold higher levels of GSH, were able to inactivate a greater proportion of cellular cisplatin, by the formation of platinum-GSH conjugates, than the CHI cells. A significant inhibition in formation of these conjugates, by BSO-induced depletion of cellular GSH (over 80%), did not, however, increase cisplatin concentration in the resistant cells. In contrast, a small increase in cisplatin concentration was observed in the sensitive cells following BSO pre-treatment. Comparison of cisplatin and tetraplatin metabolism in the SKOV-3 cells indicated that a greater proportion of the latter drug was inactivated by formation of GSH conjugates. BSO-induced depletion of cellular GSH in this cell line significantly reduced the formation of such conjugates from both drugs. However, concomitant increases in intracellular levels of reactive species were observed only after tetraplatin exposure. Our data suggest that the greater potentiation of PtIV- compared with PtII-drug cytotoxicity in the relatively resistant cell lines following 24 hr BSO pre-treatment may be caused by a differential effect of BSO on the metabolism and cellular distribution of these drugs. A BSO-induced reduction in PtII- but not PtIV-drug accumulation in these cells may also partially contribute to the differential potentiation of cytotoxicity of these drugs. PMID- 8244584 TI - Immunization with interleukin-2-secreting allogeneic mouse fibroblasts expressing melanoma-associated antigens prolongs the survival of mice with melanoma. AB - The survival of C57BL/6 mice (H-2b) bearing B16 melanoma (H-2b) was prolonged if the animals were treated solely by immunization with an interleukin-2 (IL-2) secreting allogeneic cell construct that expressed melanoma-associated antigens (MAA) along with major histocompatibility complex (MHC) class-I determinants (H 2k; RLBA-IL-2 cells). This was the case if the mice were immunized simultaneously with, or 6 days following, the injection of viable B16 cells. Under similar conditions, the survival of tumor-bearing mice immunized with an allogeneic cell construct (H-2k) that expressed MAA but did not secrete IL-2 (RLBA-ZipNeo cells), or with an allogeneic construct (H-2k) that secreted IL-2 but did not form MAA (LM-IL-2 cells), was also prolonged. However, in these instances, the period of survival was significantly shorter than that of mice immunized with the cell construct that combined IL-2 secretion with the expression of MAA. Tumor-bearing mice immunized with non-transfected LM(TK-) cells (H-2k), or irradiated B16 cells (H-2b) failed to survive longer than untreated mice. Although the survival of the treated animals was prolonged, in most instances tumor growth recurred. The recurrent tumors in mice treated with the allogeneic cell constructs formed melanin and were histologically indistinguishable from tumors in untreated mice. Cells from the recurrent tumors were resistant to further immunotherapy and to cytotoxic effector cells obtained from the spleens of mice immunized with the same cellular immunogen used initially. The injection of IL-2-secreting syngeneic B16 cells into C57BL/6 mice invariably resulted in the appearance of non-IL-2 secreting melanomas. Under similar circumstances, tumors failed to develop in C57BL/6 mice injected with IL-2-secreting, or non-secreting, allogeneic cell constructs. Thus, the expression of allogeneic antigens protected the mice from growth of the cellular immunogens. PMID- 8244585 TI - Circumvention of tamoxifen resistance by the pure anti-estrogen ICI 182,780. AB - Both primary and acquired resistance to the growth-inhibitory effects of anti estrogens (e.g., tamoxifen) limits the clinical usefulness of these drugs in the treatment of breast cancer. The new, steroidal anti-estrogen ICI 182,780 was tested for its ability to inhibit the proliferation of a tamoxifen-resistant variant of the parental MCF-7 human breast-cancer cell line. Two cell lines cloned from the MCF-7 line were used for these experiments: a tamoxifen-sensitive line, MCF 5-21, and a tamoxifen-resistant line, MCF 5-23. Compared with tamoxifen, ICI 182,780 appeared to be 150 and 1540 times more effective in inhibiting cell growth in the 5-21 and 5-23 sub-lines respectively. ICI 182,780 completely circumvented tamoxifen resistance at a concentration of (5 to 10) x 10(-9) M in this model. Based on IC50 concentrations, the 5-23 line was 22-fold more resistant to tamoxifen than the 5-21 line, but only 2-fold more resistant to ICI 182,780, reducing relative resistance by 10-fold in the resistant line. There were no differences in ER parameters between the 2 lines. ER numbers/cell were: 40500 and 34800 and the KD 0.48 and 0.15 x 10(-9) M in the 5-21 and 5-23 cells respectively. In the 5-23 cells, the concentrations of ICI 182,780 and tamoxifen resulting in a 50% inhibition of 3H-estradiol binding were 2.3 x 10(-8) M and 1 x 10(-6) M, respectively (cf. estradiol 0.89 x 10(-9) M). Thus, one potential mechanism for the increased effectiveness of ICI 182,780 may relate to the increased affinity of this drug for the estrogen receptor as compared with tamoxifen. PMID- 8244586 TI - Orthotopic implantation of mesothelioma in the pneumonectomized immune-deficient rat: a model for innovative therapies. PMID- 8244587 TI - Characteristics of the Oxford knee. Basic concepts of surface replacements. AB - Mechanical and kinematic principles are very important in knee prostheses. The use of artificial menisci is the only way to provide a kinematically free range of motion with a wide area of load transfer. This allows polyethylene wear to be minimized. However, not only the design of the prosthesis, but also the patient's underlying disease must be paid more attention. PMID- 8244588 TI - The Oxford knee. Survival analysis of joint replacement. AB - To evaluate the success of a knee prosthesis, long-term follow-up studies are needed. The best means for this is survival analysis. It would be desirable for a contribution towards the costs of these essential follow-up studies to be included in the sale price of the prosthesis. PMID- 8244589 TI - Prosthetic knee surgery in Strasbourg. AB - Knee surgery employs both joint preserving and joint replacing techniques. Even when replacement has been decided on, the further decision as to unicompartmental vs total replacement depends on many factors. A good surgeon should be able to offer all forms of treatment. PMID- 8244590 TI - The Blauth total knee endoprosthesis. Eighteen years' experience in practice. AB - A distinction must be drawn between the older constrained prostheses, in which load was transferred via metal-coated hinges, and later models using the low friction principle, where the axle is not load bearing. The latter show good long term results. The greater stability of constrained prosthesis allows earlier and less complicated mobilization even of elderly patients. PMID- 8244591 TI - Is there an ideal knee prosthesis? PMID- 8244592 TI - Normal knee joint kinematics with regard to total knee replacement. AB - Every knee has its individual envelope of motion. The joint surfaces, the ligaments and the articular capsule determine the range of motion. In walking, stability is mainly provided by the muscles. Malalignment is the usual reason for failure in knee replacement with PCA prostheses. Preoperative planning is essential for operative success. PMID- 8244593 TI - Characterization of the different prosthesis models. AB - Polyethylene is always exposed to wear. The more punctual and asymmetric the force on the surfaces, the stronger the wear. Thus, fatigue wear is more often found in unconstrained prostheses. In regard to wear problems, a prosthesis should have an axis that avoids fatigue wear and two polyethylene parts on the tibial plateau. PMID- 8244594 TI - Resolving resistances to immediacy in the group setting. AB - This article presents interventions appropriate to three forms of resistance commonly encountered in the group. One is the tendency of members to talk in the past tense instead of the present. A second is the disposition of members to dwell on the future. The third is their tendency to occupy the group with issues in their outside lives. Also mentioned are certain other common resistances to interaction that all group therapists will sometimes encounter. As important as the techniques themselves is the therapist's recognition of the need to keep groups moving forward in new and productive ways. During this is the essence of the work. PMID- 8244595 TI - The captive group: guidelines for group therapists in the inpatient setting. AB - Group therapists working with psychiatric inpatients must deal with problems unique to that setting. First, group members are usually in the hospital and the group involuntarily. They need to be recruited to attend the meetings. Second, inpatients tend to use the group as a private forum for communication with the leader. They need to be inducted into the group-as-a-whole, going beyond an exclusive focus on the leader as a need-fulfilling object. Third, members have little idea of what they are supposed to do in the group, or may be too symptomatic to observe the social mores of the group. They need to be trained as group members. Techniques for turning a "captive group" into a therapy group are discussed. PMID- 8244596 TI - Character change in group therapy. AB - The treatment of character pathology in group therapy becomes ever more relevant in the current climate of health care delivery with the mounting awareness of the importance of the long-term treatment needed for such problems. This article reviews the psychodynamic meanings of character pathology and addresses the specific ways that psychodynamic group therapy is suited to its treatment. Clinical examples are offered to illustrate how the resolution of character difficulties occurs in group therapy. PMID- 8244597 TI - Factors affecting group therapy outcome for adult sexual abuse survivors. AB - Adult female sexual abuse survivors (N = 148) participated in year-long therapy groups. Demographic, abuse history, initial symptomatology, and group process variables were examined in relationship to group completion and improvement over time. Survivors who had previously been psychiatrically hospitalized were less likely to complete group treatment. Among group completers, significant pretreatment-posttreatment changes were found on measures of locus of control, sexual problems, self-esteem, trauma-related symptomatology (TSC-33), and general psychological distress (SCL-90-R). Greater changes on the SCL-90-R were found among Caucasian women, women with more initial trauma-related symptomatology, women whose abuse included intercourse, members of groups with similar abuse histories, and women with previous psychotherapy experience. PMID- 8244598 TI - Staff-staff relations group. AB - This article describes, through the use of clinical examples, how a staff-staff relations group functions in a brief, evening, partial-hospitalization program, and how it differs from other groups within the program. It further illustrates how the appropriate use of this group allows staff to work together harmoniously and treat patients more effectively. This article also compares and contrasts this method of dealing with staff-staff and staff-patient conflict with other approaches. PMID- 8244599 TI - Open group therapy at a university counseling service. AB - A unique, open group therapy conducted at a university counseling service is described. This model differs from other approaches in that the participants "can come or not come." Method, impressions and conclusions are presented. PMID- 8244600 TI - The effect of an anti-G suit on the maximal performance of individuals with paraplegia. AB - The purpose of this study was to examine whether lower body positive pressure (LBPP) could improve maximal performance in paraplegic subjects (P). LBPP may diminish venous blood pooling (VBP) below the lesion and, therefore, support the redistribution of blood during exercise. Twelve male P subjects with complete spinal cord-lesions between T6 and T12, and 13 male able-bodied control subjects (C), performed maximal arm-cranking exercise with and without LBPP (52 mmHg) by means of an anti-gravity suit. P and C subjects had a significantly lower maximal heart rate (HRmax) with LBPP than without (P: 176 +/- 10 vs 184 +/- 7 b.min-1; C: 174 +/- 8 vs 177 +/- 8 b.min-1), whereas maximal power output (Wmax) and oxygen uptake (VO2max) remained unchanged. This suggests that LBPP provides P and C with central hemodynamic benefits. These benefits, however, did not result in improvements in Wmax or VO2max, probably due to other limitations in the complex and closely linked oxygen uptake process. During submaximal exercise the circulatory benefits were more pronounced in P than in C, probably due to the decrease of the VBP by LBPP. The results of this study suggest that LBPP appears to offer both P and C groups a central hemodynamic benefit without improving maximal performance. PMID- 8244601 TI - Plasma cyclic AMP and blood lactate responses to incremental cycling in untrained male subjects. AB - Recently, it has been suggested that epinephrine influences blood lactate and the lactate threshold during incremental exercise through a beta-adrenergic adenylate cyclase dependent mechanism. We sought to characterize the relationship between the changes in the beta-adrenergic adenylate cyclase system and blood lactate during incremental exercise indirectly through the measurement of plasma cAMP. The relationships of plasma cAMP to blood lactate levels and the lactate threshold were examined in nine untrained male subjects. Each subject performed an incremental exercise test to volitional exhaustion on an electronically-braked cycle ergometer. Although plasma cAMP was rising at the lactate threshold, it did not demonstrate the classic threshold response usually seen in blood lactate. Pairwise matched t-tests were used as a post-hoc test to determine if the successive changes in blood lactate and plasma cAMP between 21.4, 38.6, 58.7, 81.2 and 100% of VO2max were significant. Plasma cAMP was rising between 38.6% and 58.7% and between 58.7% and 81.2% of VO2max, but these changes in plasma cAMP did not reach statistical significance (p > 0.0125) with Bonferoni adjustment. The change in plasma cAMP compared to its previous value as well as the change in plasma cAMP above the resting value was not statistically significant until 81.2% of VO2max. A moderate but significant correlation was observed between blood lactate and plasma cAMP levels (r = 0.612) using blood samples obtained at each workstage in all subjects. The mean correlation between blood lactate and plasma cAMP was 0.75 (S.E. = 0.05) and ranged between 0.58 and 0.97 in individual subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244602 TI - Adaptations to six months of aerobic swim training. Changes in velocity, stroke rate, stroke length and blood lactate. AB - The purpose of this investigation was to determine how swimming velocity (SV), stroke rate (SR), stroke length (SL) and blood lactate concentration change as adaptations to six months of aerobic swim training. Subjects were trained male college swimmers (n = 8). Measurements were obtained following specially designed 400m freestyle swim tests, pre- and post-intervention. The swim test consisted of 4 x 400 m freestyle over two days. On day 1, subjects performed a maximal effort 400 m freestyle swimming trial; maximal mean velocity (Vmax) for each swimmer was calculated from this effort. On the next day, subjects were instructed to perform three 400 m freestyle swims at constant velocities equal to 85%, 90% and 95% of Vmax, respectively. Subjects rested one hour between swims. During each 400 m trial, lap time and time to complete 10 mid-pool strokes (50 m) were measured to determine SV (m.s-1), SR (stroke.min-1) and SL (m.stroke-1). Mixed arterial blood samples were taken at the end of each 400 m trial to evaluate blood lactate concentration. Results indicated that post-maximal swimming velocity (Vpostmax) increased significantly from pre-intervention measures (p < 0.05). Blood lactate concentration decreased significantly relative to SV and absolute lactate concentration following Vpostmax was significantly lower than that at Vpremax (p < 0.05). Six of seven subjects increased Vmax due to increases in SL. Mean SL during the second test was significantly higher (p < 0.05). Also, during the 400 m maximal test, SL increased significantly after sixth lap (p < 0.05). There was no significant difference between SRs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244603 TI - Usefulness of free testosterone/cortisol ratio during a season of elite speed skating athletes. AB - Free testosterone/cortisol ratio (FTCR) was used in eight elite speed skaters (five males, three females), during an eight-month season, to study the effectiveness of the hormonal parameter as a marker of the condition of overstrain and/or incomplete recovery. We did not find values lower than 0.35 x 10(-3). We confirmed the reliability of the criterion of FTCR decrease of 30% or more to indicate temporary incomplete recovery from intensive training and residual weariness and, consequently, of reduced effectiveness for competitive purposes; a formula was used in this case to calculate FTCR. When FTCR was calculated by using directly measured FT, we remarked that such threshold criterion could not be used and the significant decrease was of 18% or more. The major problem of FTCR is the limitation of its use to male athletes, although the trend found in females was similar to that presented in males. FTCR is a useful index in elite speed skaters: broader application of this index and further investigation are required to set testing times, to use direct measurement of free testosterone and, finally, to find an equivalent support for women. PMID- 8244604 TI - Validity and reliability of the Cosmed K2 instrument. AB - The purpose of this study was to measure the validity and reliability of a portable telemetry system (Cosmed K2) for measurement of oxygen consumption (VO2) during a laboratory exercise test at submaximum and maximum intensities. Twenty subjects performed three testing sessions on a treadmill on three consecutive days. Each testing session consisted of a submaximal and a maximal exercise test. On one day, VO2 was determined with the conventional Douglas bag method, and on the other two days, VO2 was determined using the Cosmed K2 system. Utilization of the Douglas bag method or the Cosmed K2 was randomly assigned. The results indicated that the Cosmed K2 instrument is reliable and valid (p > 0.05) for measurement of VO2 (l.min-1) during laboratory exercise testing at submaximum and maximum intensities. The assumption made by the K2 apparatus of a constant respiratory gas exchange ratio (R) of 1.00 did not have a significant influence on VO2 measurements. PMID- 8244605 TI - Involuntary dehydration during cricket. AB - The sweat rate, heart rate and core temperature as well as urinary volume, osmolarity, electrolyte concentration and pH of 20 cricketers were measured under cool, warm and hot conditions with wet bulb globe temperature indices of 22.1, 24.5 and 27.1, respectively. Simulated match conditions were used on the cool and warm days, while 3 bowlers were measured under actual match conditions on the hot day. The tendency for higher heart rate, sweat rate and renal conservation of water and sodium on the warm day compared with the cool day is consistent with increasing thermoregulatory stress under relatively moderate environmental conditions. The average dehydration of the three fast bowlers was -4.3% of initial body mass after only two sessions of play, on the hot day. This level of dehydration is sufficient to impair physical performance. These results suggest that the adverse effects of dehydration could be minimised if the rules of cricket were amended to allow players the opportunity to drink as desired when the environmental conditions are extreme. PMID- 8244606 TI - Effects of aerobic and anaerobic training on plasma lipoproteins. AB - We studied the effects of anaerobic and aerobic training on lipoprotein concentrations in 45 healthy untrained men. Thirty-three subjects exercised four times per week during nine weeks on a bicycle ergometer. Sixteen trained with an intensity above the anaerobic threshold (blood lactate concentration > 4 mmol.l 1) and 17 trained with an intensity below the anaerobic threshold. In addition, twelve subjects served as controls. The calculated caloric expenditure of the two training groups was similar. In all three groups, total cholesterol, total high density lipoprotein (HDL), HDL subfractions (HDL2, HDL3), and low-density lipoprotein (LDL) were measured. Training had a significant influence on HDL, HDL2, LDL/HDL, HDL2/HDL3, and cholesterol/HDL. With anaerobic training these variables changed in the opposite direction compared with aerobic training which influenced the lipoprotein profile in the desired direction. Cholesterol, HDL3, and LDL did not alter during the nine weeks of training. After nine weeks of training, the higher the blood lactate concentration during exercise (representing training intensity) was, the higher resting LDL/HDL ratio was found. The correlation between these two variables was highly significant. We conclude that training above the anaerobic threshold has no or even negative effects on blood lipoprotein profiles. Therefore, beneficial adaptations in lipoprotein profile must be achieved with moderate training intensities below the anaerobic threshold. PMID- 8244607 TI - Successful weight loss in a self-taught, self-administered program. AB - There is little evidence concerning the effectiveness of self-help materials for weight control. The purpose of this research was to evaluate a self-help weight loss program. Obese (body fat > or = 25.0%, range = 25.0-48.6%, mean +/- SEM = 36.5 +/- 1.3%) men (n = 14) and women (n = 21) were given a workbook detailing a behavior modification approach to weight loss that emphasizes self-monitoring of diet and exercise behaviors, and then sent home for 6 months to learn how to lose weight on their own. A group of 9 controls (CONT) who did not get a workbook were used for comparison. ANOVA showed that the experimental group (EXP) lost 8.1 +/- 0.9 (mean +/- SEM) kg body weight, 6.4 +/- 0.8 kg fat, and 3.9 +/- 0.6% body fat; all significant over time (p < 0.001) and different from the CONT (p < 0.0001) who showed no change in these variables. The EXP also reduced their fat intake (% of joules) from 36.1 +/- 1.0% to 27.9 +/- 1.3% (p < 0.0001), increased their carbohydrate intake from 45.7 +/- 1.2% to 50.0 +/- 1.7% (p < 0.007) and their protein intake from 16.3 +/- 0.05% to 20.7 +/- 0.7% (0 < 0.03), all of which were significantly different (p < 0.03) than the CONT who did not change. Dietary fiber increased in the EXP from 19.8 +/- 1.4 to 27.3 +/- 2.2 g/d (p < 0.001) even with a significant reduction in energy intake (11.3 +/- 0.6 vs. 8.9 +/- 0.5 Mj/d; p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244608 TI - Leisure time physical activity in the young. Correlation with low-back pain, spinal mobility and trunk muscle strength in 15-year-old school children. AB - From a population of 1503 school children, 38 15-year-old children reporting recurrent or continuous low-back pain (LBP) and 38 asymptomatic controls (34 boys and 42 girls) matched for age, sex and school class, were compared with respect to spinal mobility and trunk muscle strength. The subjects were interviewed on leisure time physical activity preceding the measurements. Of the subjects with recurrent or continuous LBP and of those completely without LBP experience 73.7% vs 42.1% reported that they pursued some physical activity two days a week or less (P = 0.006). Further, decreased spinal mobility, diminished endurance strength of the back muscles and diminished dynamic strength of the abdominal muscles were found in the group with a low frequency of activity. Mobility in children pursuing physical activity two days a week or less was decreased in lumbar extension measured by flexicurve, in forward bending, in side bending and in straight leg raising. Our results indicate that a very low frequency of physical activity in the young might be the risk factor for LBP. Pursuing sports as a leisure time activity is not harmful; on the contrary it has positive effects on spinal mobility and trunk muscle strength. PMID- 8244609 TI - Tympanometric evaluation of middle ear barotrauma during recreational scuba diving. AB - We report the first prospective evaluation of middle ear barotrauma in experienced recreational scuba divers. In this pilot study, tympanometric and otoscopic evaluations were performed daily on two experienced scuba divers engaged in multi-day repetitive diving. Middle ear pressures decreased in proportion to diving frequency, demonstrating eustachian tube dysfunction which promptly reversed upon cessation of diving Otoscopic evidence of traumatic injury to the middle ear occurred in proportion to diving frequency, and also readily reversed upon cessation of diving. Tympanic membrane compliance remained normal, often despite pronounced otoscopic abnormalities. Otologic symptoms and impairment of acuity were not observed. Tympanometry appears to be a valuable modality for the verification of middle ear hemorrhage or tympanic membrane rupture. This preliminary data should assist investigators in planning more comprehensive studies of middle ear barotrauma, including clinical trials of treatment and prophylactic interventions for this common condition. PMID- 8244610 TI - The Nottingham Adjustment Scale: a validation study. AB - The concept of adjustment to acquired sight loss is examined in the context of existing loss models. An alternative conceptual framework is presented which addresses the 'blindness experience', and which suggests that the depression so frequently encountered in those losing their sight can be understood better by recourse to cognitive factors than to psychoanalytically based theories of grieving. A scale to measure psychological status before and after rehabilitation is described, its factorial validity is demonstrated, and its validity in enabling changes to be measured. Practitioners are encouraged to adopt a similar perspective in other areas of acquired disability. PMID- 8244611 TI - Disability self-help organizations in the developing world: a case study from Malaysia. AB - Disability self-help organizations have emerged as an important element of response to the advancement of people with disabilities throughout the developing world in recent years. There are now self-help organizations in all regions of the developing world, their memberships are growing, and the scope of their activities is enlarging. This paper draws on field research in Malaysia to present an organization developed by and for people with physical disabilities as a case study of self-help action in the Southeast Asian region. This paper reviews the origins and growth of the organization, describes its current programme of activities, and offers comment about the nature and future of disability self-help in the region and its continued advancement in the developing world generally. A combination of internal factors that relate to organization and programming, and external political and social conditions is indicated as important. PMID- 8244612 TI - New home-delivered training prospects for people with disabilities. AB - The 'Home-Delivered Training Project' set out to test the viability of providing training in Information Technology to people with physical disabilities in their homes, using computers and modem links. The course included core modules on business applications software and optional modules on more specialist applications. The computer-based approach to instruction included computerized monitoring of student progress, the provision of on-line tutorial support as well as computerized test administration and marking. The 20 course participants, who lived in locations throughout Ireland, were unable to attend conventional training centres because of their limited mobility or other restrictions arising from their disabilities. They differed considerably in terms of disability type, educational qualifications and experience with computers. All but one of them completed the course successfully and were awarded certificates which are recognized in Ireland and the United Kingdom. Overall, they were very positive about the course, mainly because they could undertake it in their homes at a flexible pace to suit their learning patterns and the requirements of their disabilities. They recognized, however, that studying at home required a self disciplined study routine which was not always easy to maintain. The fact that they were awarded an internationally recognized qualification on completion of the course was an important motivating factor, as it made the course relevant in terms of their employment aspirations. PMID- 8244613 TI - Reading strategies in partially sighted children. AB - We know very little about the reading strategies adopted by children with poor vision. Here a group of children with a range of visuo-perceptual problems including defective foveal vision and impaired eye-movements are compared to a group of fully sighted children (matched on age and verbal ability). Three studies using lexical decision tasks with words (regular and irregular) and non words (legal, illegal and pseudohomophones) as well as with format distorted items, are reported. Regularity effects, as well as effects of intact and disrupted spelling units emerged but the predominant finding was that the partially sighted children though making good use of the 'visual' route in word recognition also relied on phonological mediation. The conclusions drawn were that the phonological emphasis in teaching partially sighted children was a very effective method for compensating for poor vision, and the problems of a visuo perceptual nature did not seem connected to the development of dyslexia. PMID- 8244614 TI - Mental health and functioning: a case analysis of rehabilitation patients. AB - The relationship between mental health and functioning has been inconsistently described. Here, the role of mental health in determining functioning was studied from a large multi-axial (biopsychosocial) and multi-dimensional (impairments, disabilities and handicaps) point of view. Case-analysis of rehabilitation patients indicated that inconsistent findings may be due to an incorrect linearity assumption. Results indicated that: (1) good mental health may aid the patient to benefit from rehabilitation. However, (2) it does not guarantee good future functioning, because the impact of mental health on functioning varies in interaction with other functional aspects. On the other hand, (3) poor mental health does not necessarily impede good future functioning, whereas (4) poor mental health, associated with other aspects of poor functioning, quite reliably predicts poor functioning after rehabilitation. It was concluded that single psychological factors cannot predict functioning. To improve prediction, broader models of functional assessment in rehabilitation should be used. PMID- 8244615 TI - Effects of early motor intervention in the prone position of full-term infants through the first year of life. PMID- 8244616 TI - Simple technology to promote independent activity engagement in institutionalized people with mental handicap. PMID- 8244617 TI - Delayed recording to facilitate the educational integration of people with disabilities. PMID- 8244618 TI - Community-based rehabilitation services for people with disabilities: an experimental study. PMID- 8244619 TI - The gastric groove, a specific adaptation to nutritional ontogeny in eutherian mammals. AB - In this paper grooves or groove-like structures along the lesser gastric curvature in 19 eutherian mammal species are classified according to their muscular architecture and according to mucosal differentiations. It is shown that the differentiations along the lesser curvature represent morphological and functional links between the length of lactation and weaning times, on the one hand, and the types of digestion applied by the adult animal, on the other. PMID- 8244620 TI - The effects of age and low-intensity endurance exercise on the contractile properties of single skinned fast- and slow-twitch skeletal muscle fibres. AB - Aged (25 month old) rats were subjected to a low-intensity exercise programme, consisting of 10-weeks endurance swimming. At the conclusion of training (at the age of 27 months), fresh single skinned muscle fibres were prepared from the extensor digitorum longus (EDL) and soleus (SOL) muscles of the hindlimb and activated in Ca(2+)- and Sr(2+)- buffered solutions to measure selected isometric contractile characteristics. The major fibre population of the SOL from trained aged animals demonstrated a higher threshold [Ca2+] and [Sr2+] for contraction and a reduced sensitivity to Sr2+. Few changes in contractile characteristics were observed in the EDL muscle fibres from exercised rats, except for fibre type specific changes in the steepness of the force-pCa and force-pSr relationship. The effect of ageing on the contractile characteristics of skinned muscle fibres was also investigated by comparing the data obtained from aged sedentary rats with that from adult (6-9 month old) sedentary rats. Ageing was shown not to have affected the single fibre contractile properties of the SOL, but did affect one of the two fibre populations of the EDL. This was illustrated by the higher sensitivity to Ca2+ in the aged muscle fibres with a concomitant decrease in the cooperative interactions within the thin filament during tension activation. PMID- 8244621 TI - The epigenetic influence of growth hormone on skeletal development. AB - We studied the epigenetic effect of growth hormone using mice that were transgenic for a sheep metallothionein 1a-sheep growth hormone, which was expressed beginning at 21 days postnatal age. The impact of exogenous growth hormone (GH) on various skeletal traits with special emphasis on the mandible was examined by conventional statistical analysis and finite element scaling analysis. In long bones, growth hormone enhances the proliferation rate of cartilage cells in the growth plate and should thus lead to increased lengths. Further, growth hormone is known to increase muscle mass. Our results are consistent with these developmental considerations. We found that the lengths of long bones increased in the transgenic mice compared to the control mice, while the differences in long bone width were less pronounced. In the mandible and skull, the impact of GH is most pronounced in areas of major muscle attachment, i.e., the proximal part of the mandible and the occipital and malar bones in the skull. PMID- 8244622 TI - Multiphasic allometry. AB - A multiphasic linear model is presented that describes the allometric relation between body components and permits a smooth transition from one linear segment to the next. Three applications of a diphasic linear model to allometric growth problems were used to illustrate the model. The data sets were selected because a breakpoint exists in the allometric relation. Relations between growth of the alimentary tract and of empty body weight in rabbits, growth of length and of body weight in larval-carp and growth of protein and of fat in pullets were described by a diphasic allometric model. For rabbits and larval-carp, relations were described significantly better by a diphasic instead of a monophasic (simple) allometric model. For situations where response is expected to consist of more than one linear segment, the multiphasic model is recommended. PMID- 8244623 TI - Food restriction postpones, not prevents, exercise-induced growth in hamsters. AB - Voluntary exercise during ad libitum feeding accelerates growth in mature female hamsters. If food is restricted during the exercise period, growth is suppressed, but ad libitum access to food at the cessation of exercise permits rapid catch-up growth in length and weight during retirement. To see whether the exercise induced stimulus to grow persists when food-restriction is continued during retirement, female golden hamsters were matched by weight and assigned to exercise (EX) and sedentary (SED) groups fed ad libitum and to corresponding activity groups (REST-EX, REST-SED) food-restricted to 80% of starting weight. At the end of each week for 3 weeks following retirement, one REST-EX and one REST SED group were fed ad libitum. At 21-35 days post-refeeding, mean gains in length and weight of each exercise group were greater (p < 0.05) than the gains of the corresponding sedentary group. Thus, exercise under food restriction remained an effective stimulus for acceleration of growth throughout three weeks of continued food restriction beyond termination of exercise, and ad libitum feeding permitted the delayed expression of exercise-induced catch-up growth in hamsters. PMID- 8244624 TI - Alteration of tissue levels of 13C in pigs by natural abundance 13C labeling of diets. AB - A study was conducted to determine the feasibility of using differences in the natural abundance of dietary 13C as a means of in vivo labeling. Four female pigs were fed a C3 plant diet (barley, soybean meal and alfalfa) and four were fed a C4 plant diet (corn and corn gluten meal). The pigs were continually fed the diets from 8 wks of age to maturity and through pregnancy until they had themselves produced offspring. The resulting piglets were sacrificed at various time intervals (0 to 165 days) after switching either the piglets or their mothers' to the opposite diet. Changes in the delta 13C levels in samples of milk and tissues following the diet change were monitored by isotope ratio mass spectrometry. The differences in the delta 13C content of the two diets was 12.9 parts per mil (/1000). After piglets born to a sow fed the C3 diet were switched at one day of age to a surrogate sow fed the C4 diet, tissue delta 13C levels of the piglets were nearly identical to the diet of the surrogate sow by 11 days while those switched from C4 to C3 grew more slowly and differed from the diet by 2-5/1000. When the mothers diet was switched, 15-20 days later the delta 13C in the milk had changed by only 2-3/1000, indicating that the body rather than the diet was the primary source of carbon in the milk. At 28 days after the piglets diet was switched at weaning, the tissue delta 13C had changed by 8.4/1000 (C4 to C3) and 2.9/1000 (C3 to C4). The delta 13C level in the liver changed more rapidly than in the muscle or fat and the rate of change was also influenced by the size and growth rate of the pig. PMID- 8244625 TI - Experimental studies and potential energy calculations of the blocked tetrapeptide Ac-Lys-Pro-Gly-Ile-NMA from the third loop of short-chain snake venom neurotoxins. AB - The conformational space of the tetrapeptide Ac-Lys-Pro-Gly-Ile-NMA from the beta bend present in the third loop of short-chain snake venom neurotoxins was investigated with the aid of energy calculations, resulting in the identification of an ensemble of beta-turn conformations. These results were compared with the experimentally determined conformations, as observed using NMR and CD spectroscopy. A random coil conformation of the peptide is indicated in polar hydrogen-bonding solvents. In less polar solvents the peptide backbone assumed a more rigid conformation, as reflected by the existence of at least a type II beta turn conformation. PMID- 8244626 TI - Superpotent synthetic tripeptide mimics of the mud-crab pumping pheromone. AB - Soluble pheromones released by the mud crab, Rhithropanopeus harrisii (Gould) during egg hatching cause the female crab to contract her abdomen rapidly (the pumping response). This stereotypical behavior can be induced in the laboratory by exposing egg-bearing females to solutions containing certain amino acids or peptides. Twelve amino acids exhibited response thresholds of 10(-3)-10(-10) M, the most potent (norleucine, methionine and tryptophan) having a hydrophobic sidechain. Four synthetic tripeptides of the form Gly-X-Arg, where the hydrophobic residue X was isoleucine, norleucine, methionine or phenylalanine, were superpotent agonists over a limited concentration range. Gly-Ile-Arg was a significant agonist at 10(-17) and 10(-16) M, both Gly-Nle-Arg and Gly-Met-Arg at 10(-20) and 10(-19) M, and Gly-Phe-Arg at 10(-21), 10(-20) and 10(-19) M. At the subattomolar concentration of 10(-20) M the superpotent pheromone mimics Gly-Met Arg and Gly-Phe-Arg produced not only a statistically significant increase in the relative number of pumping mud crabs but also a substantial increase in the pumping rate ratio. In contrast, at 10(-13) M the tripeptide Gly-Met(O)-Arg with an internal residue of methionine sulfoxide blocked the abdominal pumping response of egg-bearing mud crabs. PMID- 8244627 TI - Solvation of human serum albumin in aqueous alkylurea solutions. AB - Solvation of human serum albumin in aqueous solutions of urea, methyl-, N,N' dimethyl-, and ethylurea was studied by density measurements. From the densities at constant molality and constant chemical potential, the preferential solvation parameters of human serum albumin were determined. In all solutions the denaturant is preferentially bound. From preferential solvation data, Gibbs free energies of transfer from water to alkylurea solutions were calculated. Since the enthalpies of transfer were determined previously the entropies of transfer would also be obtained. Based on these and model compound data an attempt is made to identify various interactions involved in solvation. The total binding of denaturant to protein could also be calculated from preferential solvation data. The following major binding sites have been identified: ionic groups on the surface of protein molecules, peptide groups and aromatic side-chains. The correlation between the total number of sites and the number of urea molecules bound is satisfactory. PMID- 8244628 TI - Prediction of transmembrane helices from hydrophobic characteristics of proteins. AB - Membrane proteins, requiring to be embedded into the lipid bilayers, have evolved to have amino acid sequences that will fold with a hydrophobic surface in contact with the alkane chains of the lipids and polar surface in contact with the aqueous phases on both sides of the membrane and the polar head groups of the lipids. It is generally assumed that the characteristics of the aqueous parts of the membrane proteins are similar to those of normal globular proteins, and the embedded parts are highly hydrophobic. In our earlier works, we introduced the concept of 'surrounding hydrophobicity' and developed a hydrophobicity scale for the 20 amino acid residues, and applied it successfully to the study of the family of globular proteins. In this work we use the concept of surrounding hydrophobicity to indicate quantitatively how the aqueous parts of membrane proteins compare with the normal globular proteins, and how rich the embedded parts are in their hydrophobic activity. We then develop a surrounding hydrophobicity scale applicable to membrane proteins, by mixing judicially the surrounding hydrophobicities observed in the crystals of the membrane protein, photosynthetic reaction center from the bacterium Rhodopseudomonas viridis, porin from Rhodobacter capsulatus and a set of 64 globular proteins. A predictive scheme based on this scale predicts from amino acid sequence, transmembrane segments in PRC and randomly selected 26 membrane proteins to 80% level of accuracy. This is a much higher predictive power when compared to the existing popular methods. A new procedure to measure the amphipathicity of sequence segments is proposed, and it is used to characterize the transmembrane parts of the sample membrane proteins. PMID- 8244629 TI - Circular dichroism (CD) studies of antagonists derived from parathyroid hormone related protein. AB - We have undertaken a study of the structure of antagonist peptides derived from the parathyroid hormone-related protein (PTHrP) in the presence of amphiphiles using circular dichroism (CD). The results were used to gain knowledge about bioactive conformations of the peptide when bound to a membrane. The substitutions within the PTHrP-(7-34)amide sequence resulted in differences in biological activity. Structural determination by CD showed the presence of an alpha-helical structure. The antagonist activity was increased in constrained peptides in which i to (i + 4) side-chain to side-chain cyclization was used to form a lactam, [Lys13,Asp17]PTHrP-(7-34)NH2. This peptide showed increased helicity in the presence of a surfactant. Hydrophobic substitutions Leu and D-Trp at positions 11 (Lys) and 12 (Gly), respectively, in PTHrP-(7-34)NH2 resulted in increased potency, but the derivatives were not significantly more helical than the unsubstituted peptide in the presence of surfactants. The combination of the hydrophobic substitutions with the constraint of lactam formation were mutually exclusive in terms of their biological activity and their alpha-helical content. We conclude that hydrophobic substitutions contribute to an increase in binding affinity by increasing hydrophobic interactions which stabilize receptor-ligand complexes. Structural rigidification, on the other hand, increases the alpha helical content, which is important for attaining a conformation recognized by the receptor. PMID- 8244630 TI - Allyl-based groups for side-chain protection of amino-acids. AB - Allyl and allyloxycarbonyl groups are used for the side-chain protection of amino acids. The protecting groups may be selectively cleaved using the reagent HSnBu3 under palladium catalysis. The preparation of Boc and Fmoc series of protected amino acids is described. PMID- 8244631 TI - Solution structure of the toxic octapeptide, lophyrotomin. AB - Lophyrotomin is a toxic octapeptide, first isolated from larvae of the sawfly Lophyrotoma interrupta, which causes the death of cattle and sheep. It appears to act principally on the liver, however very little is known about the cellular site and mechanism of action. In the present study lophyrotomin was synthesized using solid-phase peptide synthesis, and the structure examined with two dimensional nuclear magnetic resonance (NMR) spectroscopy. Two-dimensional correlation experiments (COSY and TOCSY) enabled the assignment of many of the resonances. Conventional NOESY experiments did not produce inter-residue information, however the alternative rotating frame NOE experiment (ROESY) resulted in intra-residue alpha N, and sequential alpha N and NN NOEs, permitting the sequence-specific assignment of all resonances. The presence of few additional shortage NOEs and the absence of any long-range NOEs in the ROESY spectra indicated a lack of persistent secondary structure. The results from circular dichroism (CD) spectroscopy experiments were consistent with the NOE data, as addition of high concentrations of the denaturant urea produced no changes in the lophyrotomin CD spectrum. This conclusion was further supported by 13C spin-lattice relaxation studies, which indicated that the peptide is a flexible molecule, by examination of the alpha-carbon chemical shifts, and by amide proton exchange rate measurements. Consequently it appears that if this peptide has to adopt a well defined structure to exert its biological activity, it must do so on interaction with other molecules, such as a receptor. PMID- 8244632 TI - Acyl, pseudotetra-, tri- and dipeptide active-core analogs of insect neuropeptides. AB - Pseudopeptides of the achetakinin insect neuropeptide family were synthesized by replacing the amino acid blocks Phe-, Phe-Tyr-, and Phe-Tyr-Pro- of the active core pentapeptide Phe-Tyr-Pro-Trp-Gly-NH2 with hydrocinnamic acid, 6 phenylhexanoic acid, and both 9-phenylnonanoic and 6-phenylhexanoic acid, respectively. All four of these analogs retained myotropic activity, demonstrating that the active core could be reduced from a pentapeptide to a modified dipeptide. Most notable of these was the pseudotetrapeptide hydrocinnamyl-Tyr-Pro-Trp-Gly-NH2, which retained 70% of the potency and over 85% of the maximal activity of the parent pentapeptide. The N-terminal amino group, the phenol ring of the Tyr residue, the sulfate moiety and the Gly residue of the insect sulfakinin active core Tyr(SO3H)-Gly-His-Met-Arg-Phe-NH2 were all replaced by dodecanedioic acid. The resulting pseudotetrapeptide, dodecandioyl-His-Nle-Arg Phe-NH2, elicited myostimulatory activity. Conversely, the related acyl pseudopentapeptide azelayl-Gly-His-Nle-Arg-Phe-NH2 proved myoinhibitory. A possible explanation for these disparate biological responses is discussed. These acyl pseudopeptides are important advances towards the eventual development of stable, potent mimetic agonists and antagonists of insect neuropeptides. PMID- 8244633 TI - First characterization at atomic resolution of the C-activating groups in a peptide synthesis acid chloride, acid azide and carboxylic-carboxylic mixed anhydride. AB - The X-ray diffraction structures of three N alpha-protected, C alpha-activated MeAib derivatives are reported. They are Tos-MeAib-Cl, Tos-MeAib-N3 and Tos-MeAib O-Piv. The geometry and conformation of these classical carboxyl activating groups, which have been characterized at atomic resolution for the first time, are discussed. PMID- 8244634 TI - Discrimination against people with HIV/AIDS. PMID- 8244636 TI - WHO is recommending sweeping reforms in prisons. PMID- 8244635 TI - Tuberculosis is the leading cause of death attributable to a single infectious pathogen. PMID- 8244637 TI - Managing national nurses' associations: the UK example. AB - As the trend to restructure health systems takes momentum, there is a demand for effective management systems and managers capable of delivering quality care. Similarly, skillful management of national nurses' associations (NNAs) is essential if NNAs are to be successful in meeting their responsibilities to membership, the profession and society. Below a look at how the Royal College of Nursing, as the UK's principal professional union, has improved conditions for nurses while at the same time identifying and focusing on the "professional" issues and assuring that those who use the health services receive the highest care. PMID- 8244638 TI - Quality through management. AB - Quality assurance and performance indicators are now the order of the day. But how can nurses carry out the required orders when thy have never been sensitized to management issues in our schools of nursing. Below, Kadandara suggests a new approach to educating nurses, based on experience in Zimbabwe. PMID- 8244639 TI - Educating nurses to change their attitudes about caring for patients with HIV/AIDS. AB - Changing nurses' attitude about caring for HIV/AIDS patients is a challenge for nursing leaders, who have no choice but to promote an attractive and acceptable environment in which nurses can practice safely, compassionately and knowledgeably. Below, how nursing administrators and educators can develop strategies to meet nursing needs. PMID- 8244640 TI - Shared decision-making tested in Iowa. PMID- 8244641 TI - New, improved 'Care for Kids'. PMID- 8244642 TI - Guarding the health of student athletes. PMID- 8244643 TI - Flood-related upper respiratory infections. PMID- 8244644 TI - New coalition aims to reduce childhood injuries in Iowa. PMID- 8244645 TI - Iowa wrestlers continue detrimental weight loss practices. PMID- 8244646 TI - Cooperative Cardiovascular Project: committee updates. PMID- 8244647 TI - Recreditation through personalized CME. PMID- 8244648 TI - Ethical analysis of cases. PMID- 8244649 TI - A modified technique of delayed primary closure using a povidone iodine wick: influence on wound healing in an experimental model. AB - We report a modified technique of delayed primary wound closure using a wound wick. A rodent model was used to determine its influence on healing. After loose primary closure of two paravertebral incisions on the dorsum of male rats, a 10 cm length of ribbon gauze, soaked in either saline or 1% povidone-iodine, was inserted into the right sided wound. Wicks were soaked daily with the same solution until removal on the third post-operative day. The inclusion of a wick soaked in saline had an early transient effect on healing. A more prolonged impairment of healing was demonstrated in povidone-iodine wicked wounds and their respective controls. These data suggest that povidone-iodine impaired healing not only in the wicked wound, but also inhibited healing in the adjacent unwicked wound which was not directly exposed to the antiseptic. The observed delay in healing in povidone-iodine wicked wounds must however be balanced against its beneficial effect on wound infection rates. We suggest that further evaluation of this method of closure in contaminated wounds is warranted. PMID- 8244650 TI - Early experience with leg lengthening by callotasis and chondrodiastasis. AB - Over a four year period, 20 segments in 16 patients were treated by lengthening by callotasis or chondrodiastasis. The indications for treatment were leg length discrepancy in 13 patients and short stature in three. Patients with leg length discrepancy were all treated by femoral lengthening; the mean length gained was 4.4 cm (range 2.5 to 6.Ocm). For patients with short stature, the mean femoral length gain was 10.0 cm (9.0 to 10.5 cm), and the mean tibial length gain was 8.0 cm (6.5 to 9.0 cm). The commonest problem was pin tract infection, but this always settled with antibiotic therapy. Most other complications were also successfully dealt with, and did not compromise the outcome of treatment. At the time of review, 13 of the 16 patients said they were very happy with the result; two patients were reserving judgement until completion of treatment, and, only one thought the treatment had been of no benefit. Our initial experience, with callotasis in particular, has been that it is a highly satisfactory method of limb lengthening, has an acceptable complication rate, and involves minimal hospitalization as compared with older techniques. PMID- 8244651 TI - Correlation of peritoneal aspiration cytology with acute appendicitis. AB - Fifty-one patients in whom a clinical diagnosis of acute appendicitis had been made underwent peritoneal aspiration cytology (PAC). Thirty-six had a positive result, 11 had a negative result and aspiration failed in four cases. All patients had an appendicectomy and the histological findings were correlated with the PAC results. Thirty-five of the 36 patients with a positive PAC had histologically proven appendicitis. Seven of the 11 patients with a negative result had normal appendices. The sensitivity of PAC for acute appendicitis was 85% and the specificity was 70%. The positive predictive value was found to be 97% and the negative predictive value 60%. Peritoneal aspiration cytology is a useful aid in the diagnosis of acute appendicitis however, a negative result does not exclude this diagnosis. PMID- 8244652 TI - Fatal trauma: a five year review in a Dublin hospital. AB - The autopsy reports and clinical data of 96 patients presenting to St. James's Hospital with trauma of fatal outcome, between January 1, 1986 and December 31, 1990, were analysed. Eighty-one (84%) deaths were due to road traffic accidents and all were the result of blunt trauma. Ninety-five (99%) had an Abbreviated Injury Scale (AIS) score > or = 3 in at least one anatomical category. Cerebral trauma (41%) and shock and haemorrhage secondary to multiple injuries (40%) were the commonest causes of death. Sixty-eight (71%) patients were dead on arrival at the hospital. Of the twenty-eight deaths occurring after admission to hospital nine (32%) were potentially preventable. The mean (sem) Injury Severity Score (ISS) for those with preventable deaths was 24.1 (3.8). These data have implications for the development of trauma care facilities in Ireland. PMID- 8244653 TI - Multicentric reticulohistiocytosis--a case report. PMID- 8244654 TI - Attempted homicide in hospital. AB - Strychnine poisoning is reported in a patient who was the victim of attempted homicide in his home and in a General hospital. The diagnosis proved to be an elusive one over five months. The question of a primary psychiatric diagnosis, raised by the patient's wife proved to be an attempt by her to cover her crime. Medico-legal issues relating to the management of attempted homicide in hospital are discussed. PMID- 8244655 TI - The prevalence of microalbuminuria and associated risk factors in a population with insulin-dependent diabetes mellitus. AB - The prevalence of microalbuminuria was studied in a clinic population of patients with insulin-dependent diabetes mellitus [IDDM] with disease duration longer than 5 years. 75 patients were included in the study, 23 patients (30.7%) had microalbuminuria and 2 patients (2.7%) had macroalbuminuria. Comparison of the clinical and laboratory characteristics of patients with microalbuminuria (n = 23) to patients with normoalbuminuria (n = 50) showed no differences with respect to age, duration of diabetes, blood pressure, presence of retinopathy and glycosylated haemoglobin. The mean glomerular filtration rate (GFR) was higher in the microalbuminuric group than in the normoalbuminuric group (125 +/- 45 ml/min compared to 99 +/- 32 ml/min; P < 0.05). In a multiple regression model excluding patients with macroalbuminuria the following predictors of albumin excretion rate (AER) were identified: systolic blood pressure, glycosylated haemoglobin and GFR. Since microalbuminuria is common in Irish patients with IDDM, we suggest that AER should be determined as part of the annual routine screen. Stricter control of blood pressure and glycaemia should be considered. PMID- 8244656 TI - The role of cyclosporine on Ia antigen expression on gut epithelium in nude mice. AB - Increased expression of class II antigens (HLA-DR, human:Ia murine) on epithelial cells (EC) such as keratinocytes and gut EC suggests a role of the epithelium in the inflammatory response. It has been shown that injection of normal mouse serum (NMS) into nude mice caused an induction of Ia antigen by keratinocytes of the nude mice. The aim of the present study was to determine whether such induction could be observed in gut EC and whether it can be inhibited by cyclosporine A (CyA). Forty nude mice were divided into 4 groups of 10 each, designated A-D. Group A was injected with 0.1 ml of NMS; groups B and C were also injected with NMS and treated with oral CyA for 10 days. The dosage in group B was 30 g/ml and in group C 60 g/ml of drinking water. Group D was injected with serum of nude mice and served as a control group. Biopsies of small intestine and colon were obtained from each mouse and analyzed by indirect immunoperoxidase to identify Ia expression. Small intestine and colon EC were induced to express Ia antigen in most of the mice in groups A and B. A striking reduction in Ia expression was noted in group C. The differences between groups A and C concerning Ia expression on small intestine and colon EC were statistically significant. Our results demonstrate that the nude mouse may serve as a model for the study of Ia expression on gut EC, and that CyA can suppress the expression of Ia antigen also in the gut. PMID- 8244657 TI - Continuous therapy with intravenous prostacyclin for primary pulmonary hypertension: a bridge to heart-lung transplantation. AB - Primary pulmonary hypertension (PPH) continues to be a major problem. Vasodilator therapy has been of variable success in selected patients. Recently, with the introduction of prostacyclin (Prostaglandin I2, epoprostenon) as a potent vasodilator, several reports have shown a positive effect in primary pulmonary hypertension. We report here two patients with PPH who responded favorably to continuous intravenous therapy with prostacyclin. Both patients responded with a reduction in pulmonary vascular resistance, increase in cardiac output and dramatic improvement of right heart failure. Although therapy with prostacyclin is extremely expensive it may be used as a bridge to heart-lung transplantation in patients with end-stage PPH who do not respond adequately to alternative therapy. PMID- 8244658 TI - Home care therapy for respirator-dependent patients. AB - Our experience with 13 patients suffering from various ventilatory disorders who received mechanical ventilation at home for periods between 1 and 12 years is presented. Seven of the 13 patients were ventilated by positive pressure via tracheostomy, 3 by negative body ventilators and 3 by exufflation belt and positive pressure. Only a few hospitalizations were required due to pulmonary complications, and there was no case of sudden death occurring at home. Costs of home care for respirator-dependent patients, including the initial investment of home equipment, are much lower than the costs of long-term hospital care. A comprehensive home care program should be the preferred choice for medically stable ventilator-dependent patients. PMID- 8244659 TI - Prenatal and postnatal studies of a late infantile GM2 gangliosidosis in a family of Syrian origin: a possible B1 variant. AB - We describe late infantile Tay-Sachs disease with high residual hexosaminidase A activity in two siblings of a Syrian Druze family. The patients' leukocytes had 26% of normal hexosaminidase A activity when tested with the conventional fluorogenic substrate 4-methyl-umbelliferyl-2-acetamido-2-deoxy-beta-D glucopyranoside (4-MUG) and only about 10% when assayed with the sulfated substrate, 4-methyl-umbelliferal- beta-N-acetyl-glucosamine-6-sulfate (4-MUGS). According to the standard procedure of the heterozygote screening program (employing 4-MUG and heat inactivation), the parents were not diagnosed as an at risk couple since the father was classified as a noncarrier. However, both parents' levels were clearly within the carrier range on the basis of 4-MUGS. The unique catalytic characteristics of the patients' enzyme forward the assumption that the affected sibs are B1 variants. The parents' enzymatic levels, together with their known consanguinity, might indicate that these patients are homozygotes for the rare mutation and not genetic compounds as has been documented for most of the infantile B1 variants. To the best of our knowledge this is the first reported case of B1 variant in a child of that extraction. PMID- 8244660 TI - Computed tomography scan of the brain in pediatric neurology practice--an 8-year experience. AB - Computerized tomography scans of the brain were retrospectively studied in 1,979 children. Abnormalities were found in 45.1% of the examinations. The most common radiologic findings were enlarged ventricles (28.1% of the examinations) and enlarged subarachnoid space (14.7%). Referral diagnosis associated with high rate of radiological abnormalities included: congenital anomalies (71.7% of radiological abnormalities), cerebral palsy (70.7%), central nervous system infection (60.9%), suspicion of brain tumor (58%), and psychomotor retardation (55.6%). On the other hand, convulsive disorders (34.4% of radiologic abnormalities), learning disabilities (15.9%), and headache (11%) were associated with a relatively low rate of radiologic abnormalities. PMID- 8244661 TI - Standards of skinfold thickness and anthropometric indices in term Israeli newborn infants. AB - Anthropometric measurements including weight, length, head and mid-arm circumferences, skinfold thickness (biceps, triceps, subscapular, abdominal, suprailiac and quadriceps) were obtained from 250 singleton, full-term infants within 4 h of birth. Anthropometric indices for total body fat stores such as arm fat and nonfat areas, adiposity and ponderal indices, and weight/length ratio were calculated. These data may be helpful in determining nutritional status at birth and assessing postnatal growth of Israeli infants. PMID- 8244662 TI - Seroprevalence of antibodies against Toxoplasma gondii among two rural populations in northern Israel. AB - The prevalence of antibodies against Toxoplasma gondii was measured in two rural populations in northern Israel--Jewish kibbutz members and Arab villagers. The respective prevalences in these two populations were 22.2% and 55.8% (P < 0.001). No correlation was found between the presence of antibodies and sex, occupation, contact with cats, a history of fever and/or lymphadenopathy, eye disease, abortions or delivery of children with congenital malformations. In contrast to Jewish children who were not found to have antibodies in the first decade of life, 20.5% of Arab children tested positive. A gradual increase in the prevalence of antibodies with age was seen in both groups, with the Jews reaching a prevalence of 42.6% at age 60+ and the Arabs reaching 74% at age 40. The difference between the two groups probably stems from different eating habits, namely ingestion of raw meat and unpasteurized milk and milk products. PMID- 8244663 TI - Rupture of the uterus in labor: a review of 150 cases. AB - We analyzed 150 cases of uterine rupture in late pregnancy that occurred over an 8-year period at the State Maternity Hospital in Ankara, Turkey. The incidence of uterine rupture was 1 in 966 deliveries, and 114 uterine ruptures occurred in previously scarred uteri. Rupture of the unscarred uterus is a more catastrophic event. There is a marked difference in fetal and maternal outcome between the group with a previously scarred uterus and the group without a previous scar. Common etiological factors for uterine rupture were grandmultiparity, cephalopelvic disproportion, fetal malpresentation, and oxytocin stimulation of labor. The fetal wastage was high, a perinatal mortality of 32.2% being recorded, but 98% of mothers were saved. Hysterectomy was commonly performed in this group. Repair of the uterus should only be performed when the rupture is simple, is in the lower segment and has no sign of infection. PMID- 8244664 TI - Is Solcoderm a treatment for pigmented lesions? PMID- 8244665 TI - Hemolytic episode in a patient with Wilson's disease treated with zinc. PMID- 8244666 TI - Semen parameters among epileptic males treated with carbamazepine. PMID- 8244667 TI - Fetal effect of corticosteroid-induced maternal hyperglycemia. PMID- 8244668 TI - Mitral valve prolapse, sick sinus and Wolff-Parkinson-White syndromes: interrelationships with respect to sudden cardiac death. PMID- 8244669 TI - Prostacyclin in primary pulmonary hypertension: a bridge to heart-lung transplantation. PMID- 8244670 TI - Approach to the treatment of pigmented skin lesions. PMID- 8244671 TI - Cardiovascular disease and hormone replacement therapy: a review. PMID- 8244672 TI - Karl Landsteiner--discover of human blood types. PMID- 8244673 TI - Preface: AIDS in Israel--1993. PMID- 8244674 TI - HIV infection and intravenous drug abuse: world perspective and epidemiology in Israel. AB - As the second largest group of persons to have been infected with human immunodeficiency virus (HIV) and the main source of HIV transmission to heterosexual partners in the industrialized world, intravenous drug abusers (IVDAs) play an increasingly important role in the AIDS epidemic. The situation is particularly alarming in South and East Asia where a sharp increase in HIV seropositivity rate among IVDAs has occurred over just a few years. In Israel, the impact of HIV infection on IVDAs has remained limited with a prevalence of 2% 4.2%. In most cases, infection was contracted following travel and drug injection abroad. Israel continues to be a pre-epidemic area of HIV infection. PMID- 8244675 TI - Clinical, epidemiological and natural history study of HIV-1 infection among seropositive homo/bisexuals in Israel, 1986-92. A collaborative study in three medical centers. AB - The clinical and epidemiological aspects of HIV-1 seropositive homo/bisexuals at three AIDS screening clinics in Israel were analyzed retrospectively for the 4 year period 1986-92. Of 1,526 individuals who attended the clinics a total of 238 (15.5%) were found to be HIV-1 seropositive; 182 seropositives were followed for a mean of 3.3 (+/- 2.6) person-years. The incidence of AIDS in this cohort was 33% over 4 years, a rate of 9.8/100 person-years. Half of all HIV-1 seropositives are estimated to have been infected outside Israel. On initial presentation 144 (62%) were asymptomatic and 94 (38%) were symptomatic. CD4 levels were > 500 ml3 in 87 (36%), 200-500 ml3 in 66 (28%) and < 200 ml3 in 45 (19%) individuals. The pattern of clinical presentations was similar to that seen in the West, but mycobacterial infections were rare. A cohort of 86 homo/bisexuals with full-blown AIDS was followed for a mean of 2.2 person-years. The mortality rate in this group was 53.5% over 4 years, a rate of 26/100 person-years. Thus, HIV infection among Israeli homosexuals ceased to be a rarity as it took the pattern of AIDS in the West, with some special features such as the late presentation of patients for medical treatment and the paucity of mycobacterial diseases complicating the disease. PMID- 8244676 TI - The epidemiology of adult AIDS and HIV infection in Israel. AB - Between 1980 and 1992, 227 AIDS cases and 1,010 HIV seropositives were diagnosed in Israel, 222 AIDS cases and 965 seropositives occurring in persons aged 13 years and older. Among the AIDS cases, the predominant risk groups were male homo/bisexuals, intravenous drug abusers (IVDAs) and hemophiliacs. Among the seropositives, the predominant risk group comprised men and women originating in countries characterized by pattern II transmission; however, among nonimmigrants, male homo/bisexuals, IVDAs and hemophiliacs predominated. In recent years, there has been a gradual fall in the number of new seropositives among homosexuals and drug abusers, and the appearance of seropositive hemophiliacs has ceased. The spread of HIV infection in the indigenous heterosexual population has been negligible. PMID- 8244677 TI - Early and complete detection of HIV exposure. AB - Currently, HIV diagnosis relies on serology. Yet in groups at high risk for HIV serology is not sufficient because of the window period between infection and seroconversion. There is a growing body of reports on HIV-infected yet seronegative individuals. Some tests have been developed to identify exposure to HIV by its effect on the cells of the immune system that would differentiate following exposure to the foreign antigens. Detection, in vitro, of HIV-specific B and T cells in seronegative, at risk individuals has been reported. In only some of these individuals was an HIV infection confirmed by other methods. These new assays to detect HIV immunity enable us to identify two new groups among seronegative, at risk individuals; namely those with immunity to HIV and a detectable HIV infection (silent carriers), and those with immunity and no proof of infection. Both groups have been exposed to HIV yet are not being detected by serology. Both might hold information on other forms of HIV immunity, possibly a protective one. Thus there could be an important role for other immunological assays in early detection of HIV exposure. PMID- 8244678 TI - Host immunity and the course of HIV infection. AB - HIV infection and disease are determined to a large extent by the host's immune response. Exposure to the virus alone does not lead necessarily to long-lasting infection and the progress of HIV infection once established varies in different individuals. The development of host immunity, while either "protective" or "enhancing," will determine all these outcomes. PMID- 8244679 TI - Sexual behavior of heterosexual men and women attending an HIV testing clinic. AB - The prevalence of high risk sexual behaviors was assessed among heterosexual men (n = 154) and women (n = 109) voluntarily attending an HIV testing clinic in Jerusalem. Men were found to be involved more frequently than women in nonmonogamous relationships (66% vs. 45% respectively, P < 0.001), to have more casual partners (21% vs. 11% had three or more casual partners, P < 0.01), and to report passive oral sex (71% vs. 50%, P < 0.01) and anal sex (23% vs. 12%, P < 0.05). Condom use was not a very common practice, and was higher among men than among women (57% vs. 47% in vaginal sex, and 30% vs. 19% in oral sex respectively, NS). Appropriate counselling for this population attending the clinic is advised and urgently needed. PMID- 8244680 TI - Israeli youth and AIDS: knowledge and attitude changes among high school students following an AIDS education program. AB - A pilot project conducted in three Jerusalem schools in 1990-91 demonstrated that medical students who had received extensive training in teaching about AIDS and its prevention were able to influence the level of knowledge and attitudes on this subject among high school students. Research findings showed moderate to low knowledge levels in a sample group of 241 Jerusalem high school students on all AIDS-related questions. The average score was (mean +/- SD) 68.9 +/- 15.4, with significant differences (P < 0.01) between boys 77.2 +/- 8.6 and girls 65.9 +/- 16.4. The principal source of knowledge on AIDS among the sample population was the electronic media, 47% stating that their knowledge of HIV/AIDS came from television. After a series of four lessons in the "The Immune System and AIDS" program, there was a significant improvement in the students' knowledge. The average score rose to 78.8 +/- 13.9 with an improvement of up to 23% in the answers to the various subgroups of questions. The program succeeded in clarifying for the students the degree of personal risk from contracting HIV, but did not affect their fear of the disease--70% of the students stated that they were afraid of being ill with AIDS. The study did not examine the students' behavioral patterns. PMID- 8244681 TI - A culturally specific educational program to reduce the risk of HIV and HBV transmission among Ethiopian immigrants to Israel II: Evaluating the effect of the training program on veteran immigrant trainees. AB - We describe the evaluation of a program for training veteran Ethiopian immigrants to be health educators/cultural mediators. This educational intervention project was designed to lower the risk of transmission of HIV and HBV among the immigrants who arrived in Israel in "Operation Solomon", May 1991. The training program was designed to teach the biomedical model of HIV transmission and its prevention, and to allow the trainees to explore their feelings about people with HIV/AIDS in order to enable them to teach these issues appropriately to the general community of immigrants from Ethiopia, and to work with individuals with HIV among them. It also trained them in teaching skills and in imparting the knowledge in a culturally appropriate manner. The evaluation of changes in knowledge was done at three time points: at the beginning of a 3-day training seminar (A), its end (B) and after 7 half-day training sessions comprising the whole training program 4 months later (C). We also evaluated the changes in their attitudes and feelings about teaching these subjects and their estimate of their teaching skills at two time points (B and C). The data were collected from a written questionnaire. We also monitored their teaching through direct observation by people who spoke Amharic, and through the trainees completing a self-monitoring form. The monitoring and evaluation efforts were built into the structure of the program (including the interventions themselves). They include several complementary approaches: qualitative and quantitative, structured and semi-structured, reported and observed. PMID- 8244682 TI - HIV/AIDS among Palestinian Arabs. AB - We studied 24 HIV-positive Palestinians who presented for medical care in the West Bank, Jerusalem and the Gaza Strip between 1987 and 1992. Three individuals, all males, were detected by blood bank screening. An additional 21 individuals (12 males, 7 females, 2 infants) were diagnosed as having HIV infection. Fifteen of 24 had full-blown AIDS--11/24 (46%) were expatriates; there were 4 heterosexuals, 5 transfusion recipients and 6 homosexuals, together comprising 79% of the individuals whose risk group was known. As compared with Israelis, Palestinians presented for medical treatment at a later stage of HIV infection. We also studied the incidence of HIV infection among Palestinian blood donors between 1987 and 1992. The incidence analysis was based on HIV screening at seven blood banks of major hospitals in the West Bank, at the Makassed Hospital in East Jerusalem, at Hadassah University Hospital (Palestinian blood donors only) and at two blood banks in the Gaza Strip. Demographic features were derived from the health department of the West Bank and from the AIDS clinic at Hadassah Hospital. Approximately 50,000 Palestinians were screened at the blood banks. Only three, all males, were detected as HIV seropositive, giving an HIV overall cumulative incidence of 0.006%. This figure was similar to the incidence among 500,000 Israeli donors (0.008%; NS) during that period. Although the low HIV incidence at the blood banks is encouraging, the negligible number of HIV seropositives and the presentation at a late clinical stage points to a deficiency in AIDS tracing and in access to specialized AIDS care in the West Bank and the Gaza Strip. The small overall number of cases precludes any definite conclusion as to trends in the epidemiology of AIDS in the West Bank and Gaza. However, the receipt of contaminated transfusions and the return of infected expatriates seem to be important features of AIDS in the West Bank and Gaza. PMID- 8244683 TI - Widespread homelessness during the 1980s. PMID- 8244684 TI - Experience of family among homeless individuals. AB - The purposes of this article are to describe family experiences, both childhood and later, of homeless individuals, to propose some theoretical considerations of the influence of the family on homelessness, and to suggest programmatic structures and philosophies that consider the effect of these family experiences on the homeless experience. As part of a study of homeless people's health concerns, 40 homeless people were interviewed for 1 1/2-6 hours, each time using a semi-structured format. During the course of being interviewed, all individuals discussed experiences in their families. The commonalities among individuals' family stories were striking. As a result, experience of family was identified as an important theme in homeless people's lives. A thematic analysis of family stories was performed. As the result of the power of these stories, some theoretical considerations about homelessness that include the experience of family are proposed. Programmatic implications for psychiatric nursing's care of homeless people is discussed. PMID- 8244685 TI - Parent influences on children's self-esteem in economically disadvantaged families. AB - The purpose of this study was to test hypotheses from a transactional model of the self, related to parental influences on children's self-concepts and self esteem, within economically disadvantaged families. Thirty-nine school-age children completed self-report instruments that assessed multiple dimensions of self-perceptions, perception of parental support, importance of specific domains, and family conflict. Parents completed similar ratings of their children's specific competencies and their importance, childrearing practices, and family conflict. The results supported a model in which parents (a) influence self concepts by providing children with information on how well they perform in specific domains and which domains are important; and (b) influence self-esteem by their affective behavior. Children and parents clearly agreed on the children's competence in school, social acceptance, and behavior but not in athletics or physical appearance. Family members did not consistently agree in their importance ratings. Global self-worth was predicted by children's perceptions of parental support and perceived competence in domains that were important to the parents. PMID- 8244686 TI - Case studies of bibliotherapy with homeless children. AB - Children who experience homelessness constitute a growing population at-risk for developing mental health problems. The purpose of this study was to explore and describe the use of bibliotherapy with homeless children. A case study method of inquiry was used with drawings, audiotapes and clinical records from a convenience sample of three children living in a transitional homeless shelter. Four themes of homelessness emerged from the case studies. These were labelled "Longing for Affluence," "Escaping Violence," "Shame and Stigma," and "Making Transitions." Therapeutic stories were selected in response to the emergence of each child's own issues and the theme issues. Implications for psychiatric mental health nurses are discussed. PMID- 8244687 TI - Homeless mothers' caring for children in a shelter. AB - Homeless mothers with children are a vulnerable population that faces problems from multiple sources. Little is known about their experiences, as current homeless studies report women's data as part of the aggregate findings with men. This study sought to identify the dimension of homeless mothers' family caring in a shelter. In-depth interviews were conducted with six homeless mothers who were participants in a larger study of homeless women. Constant comparative analysis of transcribed interviews led to the identification of homeless mothers' caring behaviors for their children. These behaviors were sacrificing for children, struggling with limitations, guarding from harm, and seeking answers. This taxonomy reflects the pain and hardship these homeless families experience while living a human disaster. PMID- 8244688 TI - Hard time: the stressful nature of incarceration for women. AB - This descriptive, correlational study explored the stressful life event of incarceration for women prisoners and examined its relationship to selected health outcomes. Interviews with 55 women during their first week of incarceration and after 6 months in prison provided the data for analysis. Specific stresses of incarceration identified by the women included separation from families, worry about their children, and loss of control of their own lives. Psychological stress at time of incarceration was found to be positively related to depression and weight gain after 6 months of incarceration. Strategies to decrease the stressful nature of incarceration and improve the health status of incarcerated women are recommended. PMID- 8244689 TI - Mental health risks and problems encountered by illegal immigrants. AB - This article documents the emotional difficulties associated with the illegal Irish immigrant experience. Demographic data were collected and in-depth interviews were conducted with 17 Irish immigrants (13 had experiences as illegal immigrants and 4 were expert observers but did not have personal or first-hand experience on the topic). Content analysis of interview data documented positive and negative aspects of the illegal immigrant experience. Positive aspects included opportunity for adventure and the monetary benefit of illegal work. Negative aspects included difficulty finding satisfactory work, exploitation by employers, uncertainty about the future, vulnerability and fear over being apprehended and deported, and restrictions in numerous spheres of activity, including limited freedom to return home to visit family and friends. The findings sensitize mental health professionals to the difficulties experienced by illegal immigrants and provide direction for clinical interventions with this high-risk group. PMID- 8244690 TI - Stress and coping: the influence of racism on the cognitive appraisal processing of African Americans. AB - Individuals who experience repeated stressful events are at risk for developing physical and psychological illnesses. African Americans are an ethnic group that is exposed to a range of stressors over time, including racism which leads to discrimination. African Americans also suffer disproportionately from hypertension, cardiac disease, obesity, and drug and alcohol abuse--all illnesses that have been linked to stress. This paper describes a model to guide nursing practice, research, and education about the influence of racism on the cognitive appraisal, stress, and coping of African Americans. Lazarus and Folkman's (1984) phenomenological approach to cognitive appraisal, stress, and coping is the theoretical framework on which the model is based. PMID- 8244691 TI - Afghan refugees in California: mental health issues. AB - Refugees are a particularly vulnerable population that is at risk for mental health problems for a variety of reasons: traumatic experiences in and escapes from their countries of origin, difficult camp or transit experiences, culture conflict and adjustment problems in the country of resettlement, and multiple losses--family members, country, and way of life. Afghan refugees comprise the largest refugee population in the world, at its peak numbering more than 6 million, living mainly in Pakistan and Iran. Based on an ethnographic study of Afghan refugees in Northern California, this article describes common antecedents to and examples of mental health problems in this population, such as depression, somatic symptoms, and posttraumatic stress disorder. It reviews some of the literature on traumatized refugees and makes some suggestions to mental health providers. PMID- 8244692 TI - On the conduct of homelessness research: lessons from a qualitative study of women diagnosed with chronic mental illness. AB - This article presents a summary of the author's study of the lived experiences of homeless women diagnosed with chronic mental illness. Strategies found to be effective in the conduct of the study are highlighted. Issues and problems related to population access, sampling, data generation, and difficult situations in the field are discussed with proposed solutions. The resolution of nurse helper/nurse researcher role conflicts is addressed. PMID- 8244693 TI - A review of ecologic studies of lung cancer and indoor radon. AB - Although radon exposure is an established cause of lung cancer among underground miners, the lung cancer risk to the general population from indoor radon remains controversial. This controversy stems in part from the contradictory results of published studies of indoor radon and lung cancer, including 15 ecologic studies, seven of which found a positive association, six no association, and two a negative association. To address the misunderstanding of the indoor radon risk that has resulted from these ecologic studies, the authors discuss the general methodologic problems and limitations of ecologic studies, and the particular limitations of these 15 studies. The authors conclude that the shortcomings of the ecologic studies render them uninformative on the lung cancer risk associated with indoor radon. PMID- 8244694 TI - Video display terminals and radon. AB - Recent reports indicate that video display terminals (VDTs) can collect radon progeny from the air. This occurs especially when they are turned off and may have negative electric fields which attract positively charged radioactive dust. We have studied this phenomenon in detail and evaluated various techniques for removing the gettered radioactivity while the video display terminal is both off and on. We have evaluated what happens when the video display terminal is switched, thereby reversing the electric field near the screen. We have also studied possible inhalation effects by a video display terminal operator during field reversal and find that although some radioactivity may be released during the cycle, room air currents redistribute it into the room with no detectable levels being inhaled by users. PMID- 8244695 TI - Biospheric 129I concentrations in the pre-nuclear and nuclear age. AB - In order to detect characteristic regional differences or temporal changes of 129I concentrations in the biosphere, thyroids from humans, grazing livestock, and herbivorous wildlife species (reindeer and roedeer) were collected in various areas of the world which are not affected by reprocessing plants. For reasons of comparison, all samples were analyzed for their 129I:127I atom ratios. Human and bovine thyroids taken from the 10th region in Southern Chile (39 degrees-41 degrees South) indicated values of the 129I:127I atom ratio between 1.1 x 10(-9) and 2.0 x 10(-9) and between 1.2 x 10(-10) and 9 x 10(-9), respectively. They showed no significant increase in the concentration of biospheric 129I in comparison with that established in the pre-nuclear age. Atom ratios found in human thyroids collected in Lower Saxony (Federal Republic of Germany), which is a region not directly affected by reprocessing plants, exhibited 129I:127I values between 8 x 10(-9) and 6 x 10(-8) from February 1988 to September 1990. Thyroid glands of reindeer and roedeer as well as heather, moss, and lichen were taken from the Vilhelmina, Heby, and Gavle communes in Sweden and analyzed for 129I and 127I. All three communes were found to be seriously contaminated by fallout from the Chernobyl accident. Highest 129I:127I atom ratios between 3.5 x 10(-7) and 1 x 10(-6) were found in the Gavle commune (approximately 150 km northwest of Stockholm) where the highest 137Cs ground deposition (70-80 kBq m-2) was measured. Two soil samples taken from Krasnaya Gora and Mirny locations in Russia (approximately 200 km northeast of Chernobyl) exhibited ratios of about 1 x 10( 6). These locations showed a 137Cs ground deposition of 370 and 1,300 kBq m-2, respectively. PMID- 8244696 TI - Residual 152Eu and 60Co activities induced by neutrons from the Hiroshima atomic bomb. AB - Specific activities of 152Eu:Eu in stone samples exposed to the Hiroshima atomic bomb were determined for 70 samples up to a 1,500-m slant range from the epicenter. The specific activities of 60Co:Co were also determined for six samples near the Hiroshima hypocenter. First, the 152Eu data were investigated to find out the directional dependence of neutron activation. Directional anisotropy was not definite; however, there was an indication that the activation in the west-southwest was lower than in other directions. Second, measured 152Eu and 60Co radioactivity data were compared with activation calculations based on DS86 neutrons. It is clearly shown that the measured data are lower than the calculation near the hypocenter and vice versa at long distances beyond 1,000 m. The calculated-to-measured ratios of 152Eu are 1.6 at the hypocenter, 1.0 at approximately 900 m, and 0.05 at a 1,500-m slant range. Present results indicate that systematic errors exist in the DS86 neutrons concerning the source-term spectrum, neutron transport calculations in air, and/or activation measurements. PMID- 8244697 TI - Estimate of excess uranium in surface soil surrounding the Feed Materials Production Center using a requalified data base. AB - A conservative estimate of the excess total uranium in the top 5 cm of soil surrounding the former Feed Materials Production Center was made using a data base compiled by the International Technology Corporation in 1986, and the requalification of that data base was completed in 1988. The results indicate that within an area of 8 km2, extending 2 km both northeast and southwest of the Feed Materials Production Center, the uranium concentration is between 2 and 5 times greater than average natural background radiation levels. More than 85% of this excess uranium is deposited within 1 km of the site boundary. The presence of any excess uranium outside of this area is indistinguishable from the natural background contribution. PMID- 8244698 TI - A predictive model for the behavior of radionuclides in lake systems. AB - This paper describes a predictive model for the behavior of 137Cs in lacustrine systems. The model was tested by comparing its predictions to contamination data collected in various lakes in Europe and North America. The migration of 137Cs from catchment basin and from bottom sediments to lake water was discussed in detail; these two factors influence the time behavior of contamination in lake water. The contributions to the levels of radionuclide concentrations in water, due to the above factors, generally increase in the long run. The uncertainty of the model, used as a generic tool for prediction of the levels of contamination in lake water, was evaluated. Data sets of water contamination analyzed in the present work suggest that the model uncertainty, at a 68% confidence level, is a factor 1.9. PMID- 8244699 TI - Benefits to a life insurance company from providing radon tests for clients. AB - If a life insurance company provided free radon tests to clients, clients' life expectancies would be extended and profits would thereby be increased. This effect is quantified and it is found that the direct monetary benefits to the company could be substantial. Several subsidiary advantages are also discussed. PMID- 8244700 TI - Assessment of the natural radioactivity in the Brazilian state of Rio Grande do Norte. AB - Fifty-one soil samples were collected to a depth of 30 cm in the central and eastern region of the Brazilian state of Rio Grande do Norte. Concentrations of background radionuclides in soil samples were determined by gamma ray spectrometry with an HPGe detector. The average concentrations of 226Ra, 232Th, and 40K in the surveyed soils were 29.0 +/- 19.4 Bq kg-1, 46.6 +/- 36.2 Bq kg-1, and 677.8 +/- 434.9 Bq kg-1, respectively. The bedrock of Santana do Matos (Rio Grande do Norte) showed fairly high radioactivity (90 Bq kg-1 of 226Ra, 285.6 Bq kg-1 of 232Th, and 1,414 Bq kg-1 of 40K). Radiological measurements carried out in Santana do Matos with a hand-held scintillometer revealed external gamma radiation ranging from 200-330 nGy h-1 in the down-town area. Higher values were recorded in the surroundings of that town. PMID- 8244701 TI - Normal concentration and excretion ratio of uranium in serum of normal individuals in India. AB - This paper reports the normal concentration of uranium in the blood serum of 12 normal volunteers from the Bombay, India area who had no occupational exposure to uranium. Serum samples were analyzed by neutron activation, radiochemical separation, and gamma counting. The mean and median uranium concentrations obtained in the present study are 14 and 13 ng L-1, respectively. The median value was used to obtain the excretion ratio of 41% for uranium excreted in urine compared to that present in the blood serum pool. PMID- 8244702 TI - A method to obtain subject background for low-level in-vivo measurements of the head. AB - Background for an in-vivo whole-body counter measurement is commonly obtained by averaging the counts of one or more matched (size, weight, gender) control subjects who have not been exposed to the radionuclide of interest. This limits the statistical significance to the time of the count, generally 30 min to 1 h when counting people. To obtain a human background with a small associated counting error in this manner would require counting the matching control subjects for a period much longer than they could tolerate. Also, it may not be possible to find a matching control subject who has not been exposed to the radionuclide of interest, as in the case of 210Pb, a nuclide that is normally present in air, food, and water, and to which everyone has been exposed all of their lives. This paper presents a method developed at New York University Medical Center, Nelson Institute of Environmental Medicine to obtain a matching background for any subject, for use in the measurement of 210Pb in the skull, in vivo. It is obtained using a head phantom containing known amounts of potassium and calcium. The phantom, surrounded by three phoswich detectors in the same position as the subject's head, is counted repeatedly for long periods. Although it has been used to obtain "head" backgrounds only in the 210Pb spectral region, the method should be applicable to other regions of the low-energy phoswich spectrum. PMID- 8244703 TI - Performance test of PDM-101 electronic pocket dosimeters using a 60Co source. AB - The performance tests of one type (named PDM-101) of silicon semiconductor dosimeter were carried out by using a 60Co point source (approximately 500 kBq). The dosimeter responds to dose equivalents on the order of 0.02 microSv although measured data show a large error for values < 0.08 microSv. The deviation of measured dose equivalents from calculated ones was < 10% in the region > or = 0.08 microSv. The deviation of measured dose equivalent rates from calculated ones was generally < 10%. The average value of measured dose equivalents for 31 directional exposures without underside exposure was 0.92 +/- 0.06 of the calculated value. The difference of measured data for the front and back exposure was < 10%. The background and noise level of the dosimeter was 0.092 +/- 0.005 microSv h-1. All measured data showed that dosimeter response for radiation was symmetric between the front (left) and the back (right) sides of the dosimeter within 10% tolerance. PMID- 8244704 TI - Monitoring 90Sr contamination in terms of 131I contamination in imported food. AB - This paper describes a method for monitoring 90Sr contamination in terms of 131I contamination in food imported from a country with a recent nuclear power plant accident. For imported food, the guideline levels set by the Codex Alimentarius Commission will be applied, which requires that the contamination in food by beta emitters (typically strontium) be monitored. However, the measurement of strontium contamination cannot be made within a reasonably short time as it requires rather time-consuming chemical extraction and sample preparation procedures in advance. We propose here to use the contamination level of 131I in these foodstuffs as indicators of 90Sr contamination levels. Possible accident scenarios are employed to estimate the abundance ratio of 131I and 90Sr deposited on soil and intercepted by plants. Using a dynamic food chain model, we can calculate their relative activities in the food products which, when combined with the 131I levels present, give the theoretical 90Sr levels present. PMID- 8244705 TI - An augmented audit program for assuring radiation safety during radiographic examination operations. AB - Auditing a gamma radiography program is required as part of the authorizing license. Checklists and cursory reviews are the typical approach to addressing program requirements. A more proactive approach is recommended. The audit program described was prepared for a specific set of operating conditions but can be applied to any given program. Improvements in the effectiveness of the radiography safety program can be made with additional examination and emphasis on direct observation of licensed activities. PMID- 8244706 TI - Preface: tritium dosimetry, health risks, and environmental fate. PMID- 8244707 TI - Special issue: Tritium dosimetry, health risks, and environmental fate. PMID- 8244708 TI - Overview of tritium: characteristics, sources, and problems. AB - Tritium has certain characteristics that present unique challenges for dosimetry and health-risk assessment. For example, in the gas form, tritium can diffuse through almost any container, including those made of steel, aluminum, and plastics. In the oxide form, tritium can generally not be detected by commonly used survey instruments. In the environment, tritium can be taken up by all hydrogen-containing molecules, distributing widely on a global scale. Tritium can be incorporated into humans through respiration, ingestion, and diffusion through skin. Its harmful effects are observed only when it is incorporated into the body. Several sources contribute to the inventory of tritium in our environment. These are 1) cosmic ray interaction with atmospheric molecules; 2) nuclear reactions in the earth's crust; 3) nuclear testing in the atmosphere during the 1950s and 1960s; 4) continuous release of tritium from nuclear power plants and tritium production facilities under normal operation; 5) incidental releases from these facilities; and 6) consumer products. An important future source will be nuclear fusion facilities expected to be developed for the purpose of electricity generation. The principal health physics problems associated with tritium are 1) the determination of the parameters for risk estimation with further reduction of their uncertainties (e.g., relative biological effectiveness and dose-rate dependency); 2) risk estimation from complex exposures to tritium in gas form, tritium in oxide form, tritium surface contamination, and other tritium contaminated forms, with or without other ionizing radiations and/or nonionizing radiations; 3) the dose contributions of elemental tritium in the lung and from its oxidized tritium in the gastrointestinal tract; 4) prevention of tritium (in oxide form) intake and enhancement of tritium (oxide form) excretion from the human body; 5) precise health effects information for low-level tritium exposure; and 6) public acceptance of tritium leakage and waste disposal from reactors and fuel reprocessing plants. PMID- 8244709 TI - Tritium sampling and measurement. AB - Current methods for sampling and measuring tritium are described. Although the basic techniques have not changed significantly over the last 10 y, there have been several notable improvements in tritium measurement instrumentation. The design and quality of commercial ion-chamber-based and gas-flow-proportional counter-based tritium monitors for tritium-in-air have improved, an indirect result of fusion-related research in the 1980s. For tritium-in-water analysis, commercial low-level liquid scintillation spectrometers capable of detecting tritium-in-water concentrations as low as 0.65 Bq L-1 for counting times of 500 min are available. The most sensitive method for tritium-in-water analysis is still 3He mass spectrometry. Concentrations as low as 0.35 mBq L-1 can be detected with current equipment. Passive tritium-oxide-in-air samplers are now being used for workplace monitoring and even in some environmental sampling applications. The reliability, convenience, and low cost of passive tritium-oxide in-air samplers make them attractive options for many monitoring applications. Airflow proportional counters currently under development look promising for measuring tritium-in-air in the presence of high gamma and/or noble gas backgrounds. However, these detectors are currently limited by their poor performance in humidities over 30%. PMID- 8244710 TI - Metabolism and dosimetry of tritium. AB - This document was prepared as a review of the current knowledge of tritium metabolism and dosimetry. The physical, chemical, and metabolic characteristics of various forms of tritium are presented as they pertain to performing dose assessments for occupational workers and for the general public. For occupational workers, the forms of tritium discussed include tritiated water, elemental tritium gas, skin absorption from elemental tritium gas-contaminated surfaces, organically bound tritium in pump oils, solvents and other organic compounds, metal tritides, and radioluminous paints. For the general public, age-dependent tritium metabolism is reviewed, as well as tritiated water, elemental tritium gas, organically bound tritium, organically bound tritium in food-stuffs, and tritiated methane. PMID- 8244711 TI - Microdosimetry of tritium. AB - Microdosimetric aspects of tritium radiotoxicity are discussed. The level of coincidence of tritium locations and of radiation-sensitive sites does not appear to be decisive for relative effectiveness of tritium exposures. The actual target structure and dimensions seem to be of more importance. Particularly efficient germ cell destruction by chronic exposures to tritiated water is unlikely to be explainable on the basis of classical principles of microdosimetry. The phenomenological approach of investigating biological response functions may be a useful alternative for the interpretation of unexpected experimental results, especially when combined with bidimensional microdosimetry. The latter allows for clustering of nanometer sites within a target of cellular dimensions. PMID- 8244712 TI - Tritium radiobiology and relative biological effectiveness. AB - During the past decade, a large number of radiobiological studies have become available for tritium--many of them focusing on the relative biological effectiveness of tritium beta rays. These and previous studies indicate that tritium in body water produces the same spectrum of radiogenic effects (e.g., cancer, genetic effects, developmental abnormalities, and reproductive effects) observed following whole-body exposure to penetrating radiations such as gamma rays and x rays. However, tritium beta rays are of greater biological effectiveness than gamma rays and x rays. For example, tritium in the oxide form is about 2 to 3 times more effective at low doses or low dose rates than gamma rays from 137Cs or 60Co. When tritium is bound to organic molecules, relative biological effectiveness values may be somewhat larger than those for tritium in oxide form. Tritium administered to animals or to cells in vitro as tritiated amino acids results in relative biological effectiveness values that appear similar to those obtained for tritium in oxide form; however, if administered as tritiated thymidine, the relative biological effectiveness values appear to be about two-fold higher. It is clear from the wealth of tritium data now available that relative biological effectiveness values for tritium beta rays are higher than the quality factor of unity generally used in radiation protection. PMID- 8244713 TI - Tritium risk assessment. AB - Estimates of the health risks in humans from low-level exposure to tritium are presented. The health risks considered are those for cancer, genetic effects, and developmental abnormalities from exposures in utero. Because direct risk information for these effects is not available from human exposures to tritium, the following approach was used. Excess risks for the effects given following low level exposure to x rays or gamma rays were estimated from available human epidemiological data using appropriate dose-rate effectiveness factors. These human-risk estimates for low-level x rays or gamma rays were then multiplied by the appropriate best-estimate relative biological effectiveness for tritium, taking into account differences in effectiveness of comparison radiations. The resultant lifetime risk coefficients for low-level exposure to tritiated water are as follows. For cancer mortality, the most probable risk (50th percentile) is 81 x 10(-6) mGy-1 and the 90% confidence interval is 38 to 185 x 10(-6) mGy-1. For genetic effects in the first generation after exposure the risk is 7.9 x 10( 6) mGy-1, with a 90% confidence interval of 3.8 to 16.3 x 10(-6) mGy-1. For developmental effects from low-level tritiated water exposures in utero, the risk is uncertain but is estimated to be < 400 x 10(-6) mGy-1. The risks from exposure to organically bound tritiated molecules are estimated to range from values that are similar to those for tritiated water to about a factor of 2 higher. PMID- 8244714 TI - Tritium transport and cycling in the environment. AB - The transport and cycling of tritium in the environment can be understood in terms of the role of hydrogen in the environment. Physical and chemical isotopic effects, while present in some transport mechanisms and chemical reactions, are not important factors in environmental tritium dynamics. In addition, because of the role of hydrogen in metabolism and its ubiquitousness in the environment, organisms have not evolved mechanisms to accumulate or concentrate hydrogen or its isotopes in food chains. Therefore, biomagnification of tritium is not a factor in food chain transfer. The lack of significant isotopic effects or biomagnification means that tritium transport and cycling in the environment can be predicted on the basis of the transport processes, hydrogen content, and chemical transformation of hydrogen and its compounds in the environment. PMID- 8244715 TI - Organically bound tritium. AB - Tritium released into the environment may be incorporated into organic matter. Organically bound tritium in that case will show retention times in organisms that are considerably longer than those of tritiated water which has significant consequences on dose estimates. This article reviews the most important processes of organically bound tritium production and transport through food networks. Metabolic reactions in plant and animal organisms with tritiated water as a reaction partner are of great importance in this respect. The most important production process, in quantitative terms, is photosynthesis in green plants. The translocation of organically bound tritium from the leaves to edible parts of crop plants should be considered in models of organically bound tritium behavior. Organically bound tritium enters the human body on several pathways, either from the primary producers (vegetable food) or at a higher tropic level (animal food). Animal experiments have shown that the dose due to ingestion of organically bound tritium can be up to twice as high as a comparable intake of tritiated water in gaseous or liquid form. In the environment, organically bound tritium in plants and animals is often found to have higher specific tritium concentrations than tissue water. This is not due to some tritium enrichment effects but to the fact that no equilibrium conditions are reached under natural conditions. PMID- 8244716 TI - Tritium radiobiological effects in mammals: review of experiments of the last decade in Russia. AB - This review briefly describes techniques and basic results of experimental investigations in mice and rats on metabolism, dosimetry, and radiobiological effects of tritium oxide and some tritiated biogenic compounds (glucose, amino acids, and nucleosides) during the last 10 to 15 years in Russia. The content of water in tissue cells of mammals is shown to be 15 to 40% less than in whole tissue. The kinetics of tritium incorporation from oxide (HTO) and its retention in DNA of hemopoietic tissues were studied. The contribution of bound tritium to dose strongly depends on the chemical form of tritium and reaches 90% when labeled L-lysine is injected. Specific features of the action of HTO on hemopoietic tissue were investigated in tests of damage and repair of DNA, induction of chromosome aberrations in cells, content of nucleic acids, kinetics of cell populations, immunity parameters, carcinogenesis, decrease of life span, induction of dominant lethal mutations in germ cells in male mice, and reciprocal translocations in mouse spermatogonia. According to these tests, the radiobiological effects of tritium beta radiation in the form of oxide is 2 to 6 times higher than for gamma radiation of 137Cs. The frequency of dominant lethal mutations induced by labeled lysine, thymidine, and deoxycytidine is 3 to 12 times higher than those induced by equal HTO activity. The results of these investigations are used to standardize HTO and the various biogenic compounds of tritium, improve techniques of indirect dosimetry, provide medical aid to personnel, and estimate population risk. PMID- 8244717 TI - Double whammy own goal. PMID- 8244718 TI - Homelessness. Who likes to B&B beside the seaside? PMID- 8244719 TI - HVA fights 'probationary' F grades. PMID- 8244720 TI - Marketing. I. Lessons in talking business. AB - The Welsh Office has invested thousands of pounds in a national training programme to help community nurse managers and field staff market their services to GPs, commissioning health authorities and the public. Catherine Jackson reports on the genesis of the project and the benefits to health visitors in the field. PMID- 8244721 TI - Marketing. 2. Hook, line and sinker. PMID- 8244722 TI - Marketing. 3. Contract culture. AB - Health visiting and school nursing services are now governed by the contracts agreed between health authorities and trusts under the purchaser/provider split. Jean Rowe describes how purchaser and provider do business in the health care market. PMID- 8244723 TI - Group work. Cope street revisited. AB - 45 Cope Street is a preventive health project working with pregnant women and young mothers aged between 16 and 25 years and their children in the inner city of Nottingham. A beacon of innovative health visiting practice, it has recently passed the fifth anniversary of its opening. Team leader Ann Rowe describes their philosophy and practice. PMID- 8244724 TI - Thalassaemia: 1 of 3. Establishing basic awareness. AB - Thalassaemia has until recently received little attention and consequently is not widely understood by health professionals, write Simon Dyson, Verna Davis and Rafeya Rahman. In the first of three articles they describe the illness and its treatment and the potential role for health visitors in supporting children with the condition and their families. They conclude with a brief questionnaire to test thalassaemia-awareness levels. PMID- 8244725 TI - Promoting physical activity in schools. AB - Neil Armstrong, director of the Coronary Prevention in Children Project, argues for a comprehensive programme for promoting children's physical activity. The project's survey of adult coronary risk factors in British children revealed a worryingly low level of physical activity among British schoolchildren. Schools are ideally placed to encourage children to take physical exercise, he writes, but parental role models also play an important part. PMID- 8244726 TI - Teenage screening in a general practice setting. AB - Concern about the apparent lack of health awareness among local teenagers prompted practice nurse Anne Campbell and medical receptionist Sheila Edgar to launch a health promotion programme aimed at this age group. Here they report the success of the 'Teenscreen' project. PMID- 8244727 TI - Postnatal support. Listening: an under-valued health visiting skill. AB - The experience of being listened to can be a turning point in people's lives, particularly in the early months of parenthood, write Shona Reed-Purvis and Sally Dakin. This is a time when health visitors are ideally placed to offer emotional support. But training may be necessary to help the practitioner move beyond the traditional directive role engendered by old-style nurse training and expected by many clients, and to encourage the client to take control. Here they describe a training programme developed at the request of Oxford City health visitors to help them use their skills more effectively. PMID- 8244728 TI - Profiling. Less impertinence, more partnership. AB - Health profiling is a central aspect of health visiting practice, writes Ian Bell. Here he describes the development of a profiling tool for use with all age groups to enable accurate assessment of clients' health needs. PMID- 8244729 TI - A Victorian early childhood: the parental record of a baby born in 1892. AB - Histories of Victorian child rearing practices have usually concentrated on the most materially deprived sections of society: the communities living in conditions such as necessitated the employment of the Salford sanitary inspectors, writes Patience Karseras. These extracts from the diary of her grandmother chart the milestones and significant events in the early life of a baby--her father--born to a prosperous middle class family. PMID- 8244730 TI - Punishing the poor. PMID- 8244731 TI - TUPE. Safe transfer. AB - The NHS and Community Care Act protected the pay and conditions of NHS staff transferred from health authority employment to trust. But with the advent of GP fundholding and moves by health care purchasers to 'play' the NHS internal market, HVA members will need the protection of TUPE: the Transfer of Undertakings (Protection of Employment) regulations 1981, as Roger Kline explains. PMID- 8244732 TI - The mathematics of community care. PMID- 8244733 TI - An overbalanced response by industry? PMID- 8244734 TI - Flying a flag for standards. PMID- 8244735 TI - Abortion. Clinics prepare for autumn protests. PMID- 8244736 TI - Immunisation. Facts and fiction. AB - Immunisation is widely acknowledged to be one of the most effective public health interventions of this century. Very few parents positively refuse immunisation for their child, but their objections are fuelled by national media and professional journal coverage of the counter arguments. Helen Bedford reviews the case for immunisation. Health professionals need to be able to discuss the issues from an informed, balanced base if they are to advise those parents who believe it is of no benefit and may even harm their child, she writes. PMID- 8244737 TI - An update on headlice. PMID- 8244738 TI - Health visiting. Principles in practice: a re-affirmation. AB - Sheila Twinn suggests that an ad hoc development of health visiting practice in recent years has contributed to the current confusion and uncertainty over its future role. Here she outlines a new framework for practice based on a re affirmation of the four principles of health visiting and on reflective practice. PMID- 8244739 TI - Measuring standards in community practice teaching. AB - Doreen Langford describes how a group of community practice teachers (CPTs) audited their work. Nine professional standards for the training of health visitor students were agreed and success in meeting those standards assessed using Donabedian's model of evaluation. PMID- 8244740 TI - Pros and cons of family credit. AB - Family credit (FC) is the best thing since sliced bread--at least that is what the government would like potential claimants and advisers to believe. Certainly family credit can assist some families on low pay but there are also a number of problems with the benefit, as Beth Lakhani of the Child Poverty Action Group explains. PMID- 8244741 TI - Health promotion. Pitching the message at men. AB - Haringey health services beamed its safer sex message into millions of homes all over Britain. It was splashed across almost every national newspaper and got saturation coverage in the local press and on local radio. So how did they do it? Cath Jackson reports on a whole new ball game in health promotion. PMID- 8244742 TI - Sharing drink-wisdom. PMID- 8244743 TI - Evaluating community practice teaching in Doncaster. AB - Managers in Doncaster health district were concerned to ensure community practice teachers give a high quality standard of teaching to students. June Evans and Dorothy Sedgwick describe the process of introducing audit to the community practice teaching of health visitor and district nurse students. PMID- 8244744 TI - Less means less. AB - The UKCC's proposed reform of community nurse education (PREP) will not adequately prepare health visitors for their demanding role in community health care, health visitors fear. Frances Appleby and Anne Robotham present the views of leading health visitor educationalists, while (right) 41 health visitors from Kent make their views. PMID- 8244745 TI - Misrepresenting milks: the industry responds. PMID- 8244746 TI - Laser safety eyewear. AB - In spite of repeated warnings about laser safety practices, as well as the availability of laser safety eyewear (LSE), eye injuries continue to occur during use of surgical lasers, as discussed in the Clinical Perspective, "Laser Energy and Its Dangers to Eyes," preceding this Evaluation. We evaluated 48 models of LSE, including goggles, spectacles, and wraps, from 11 manufacturers. The evaluated models are designed with absorptive lenses that provide protection from CO2 (carbon dioxide), Nd:YAG (neodymium:yttrium-aluminum-garnet), and 532 (frequency-doubled Nd:YAG) surgical laser wavelengths; several models provide multiwavelength protection. (Refer to ECRI's Product Comparison System report on LSE for specifications of other models.) Although most of the evaluated models can adequately protect users from laser energy--provided that the eyewear is used -many models of LSE, especially goggles, are designed with little regard for the needs of actual use (e.g., adequate labeling, no alteration of color perception, sufficient field of vision [FOV], comfort). Because these factors can discourage people from using LSE, we encourage manufacturers to develop new and improved models that will be worn. We based our ratings primarily on the laser protection provided by the optical density (OD) of the lenses; we acknowledge the contribution of Montana Laser Optics Inc., of Bozeman, Montana, in performing our OD testing. We also considered actual-use factors, such as those mentioned above, to be significant. Among the models rated Acceptable is one whose labeled OD is lower than the level we determined to be adequate for use during most laser surgery; however, this model offers protection under specific conditions of use (e.g., for use by spectators some distance from the surgical site, for use during endoscopic procedures) that should be determined by the laser safety officer (LSO). LSE that would put the wearer at risk are rated Unacceptable (e.g., some models are not properly or clearly labeled, have measured ODs that are not adequate for protection, or significantly restrict the wearer's FOV); also, LSE with side shields that do not offer adequate protection from diffuse laser energy are rated Unacceptable. Those models that offer adequate protection for surgical applications, but whose measured OD is less than their labeled OD, are rated Acceptable--Not Recommended; if the discrepancy is great, they are rated Unacceptable. Those models whose labels were removed during cleaning are rated Conditionally Acceptable.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8244747 TI - Healthco disposable saliva ejectors: dislodged tips. PMID- 8244748 TI - Puritan-Bennett Companion 360 oxygen regulator: disconnected flow-control knob. PMID- 8244749 TI - Electronic equipment mounted on IV poles. PMID- 8244750 TI - Disposable breathing circuits for portable ventilators. PMID- 8244751 TI - Investigating device-related "burns". AB - In this article, we discuss a procedure for investigating skin injuries sustained by patients in the operating room and special care areas of the hospital. Skin injuries are often mistaken for burns, and medical devices are immediately blamed as the cause; however, such a hasty conclusion can overlook the actual cause of the injury and delay the implementation of measures to prevent future occurrences. We describe a thorough investigation process for uncovering the real cause of the injury and include a questionnaire to facilitate the investigation. We also list potential causes of accidental skin injuries and present two investigation case studies. PMID- 8244752 TI - Baxter PowerCut Arthroscopic Shaver System handpiece not modified after recall (patient suffers gas embolic brain death). PMID- 8244753 TI - The gene for bovine interferon gamma (IFNG) maps to the q22-q24 bands of chromosome 5 in cattle. AB - A approximately 700 bp bovine IFNG cDNA probe was used for the chromosomal localization of the IFNG gene in cattle using in situ hybridization technique. This locus has previously been assigned to chromosome 5 by analysis of a somatic cell hybrid panel. The in situ hybridization results obtained in the present study allow us to map the locus at the 5q22-q24 bands. Relative location of the IFNG gene and the class II cytokeratin and HOX3 gene clusters in cattle and humans is discussed. PMID- 8244754 TI - Effect of the O chromosome of Drosophila subobscura on abdominal bristle number. AB - The role of the O chromosome of D. subobscura on the control of abdominal bristle number was analyzed. The study was performed using four strains derived from four lines selected for high (H1 and H2) and low (L1 and L2) bristle number. In all the lines, the factors for abdominal bristle number carried on the O chromosome showed additive effects and diminished the number of abdominal bristles. The negative effect of the O chromosomes from low selection lines was found to be stronger than the negative effect of the O chromosomes from high selection lines. PMID- 8244755 TI - Zoogeography of the chromosome 1 HSR in natural populations of the house mouse (Mus musculus). AB - A polymorphism of the central part of chromosome 1 has been described from natural populations of the house mouse (Mus musculus). The region shows up as a C band-positive homogeneously staining region (HSR) under the light microscope. M. m. domesticus mice carry single band HSRs, whereas M. m. musculus animals have double band HSRs. HSR size variations have been described in both subspecies. The frequency of the HSR chromosome 1 in populations varies from 4% to 81%, but none of the large samples examined consisted only of homozygotes. In the subspecies M. m. domesticus, HSRs were found in North Africa and Western Europe, mainly in the hilly regions of Southern Germany and Switzerland. Localities with double HSRs are distributed all over the area of M. m. musculus. Based on the population data presented and DNA similarity of different HSRs, the origin and distribution of HSR chromosomes in the house mouse are discussed. PMID- 8244756 TI - Expression of the dihydroflavonol reductase gene in an anthocyanin-free barley mutant. AB - The barley gene encoding dihydroflavonol-4-reductase (DFR) was delivered by micoprojectile bombardment into leaf sheath tissue of the anthocyanin-free barley mutant ant 18-162, a mutant which lacks DFR activity-probably because of a missense mutation in the structural gene for DFR. The delivered gene complemented the mutation, as evidenced by the synthesis of anthocyanin in individual leaf sheath cells of the bombarded tissues. Pigment synthesis appeared two days after gene delivery and both the number of pigmented cells and the intensity of pigmentation increased over the following days. Depending on the physiological condition of the host plants, up to 15 pigmented cells per 10 tissue segments were detected. These results demonstrate that the Ant 18 gene of barley encodes dihydroflavonol-4-reductase. A series of gene constructs encoding DFR were expressed in the anthocyanin-free mutant tissue. The genomic clone complemented the mutation whereas an equivalent plasmid with all introns deleted did not. The highest number of pigmented cells was obtained using plasmids containing the DFR coding sequence interrupted by intron 1 of the genomic clone, indicating that the presence of an intron stabilizes the DFR message. PMID- 8244757 TI - Localization of the ceruloplasmin (CP) gene to the q32-q33 bands of chromosome 13 in pigs by in situ hybridization. AB - Ceruloplasmin (CP) is a copper-binding protein in vertebrate plasma. In the present study, the porcine ceruloplasmin gene was localized to the 13q32-q33 bands by in situ hybridization, using a human CP cDNA probe. This confirmed the localization of the porcine linkage group V to chromosome 13. The CP locus is closely linked to the transferrin locus in pigs. Their relative physical order on chromosome 13 is discussed. Comparisons are made with the order of these two loci in other mammalian species. PMID- 8244758 TI - Simultaneous detection of high-resolution R-banding and fluorescence in situ hybridization signals after fluorouracil-induced cellular synchronization. AB - A method for simultaneous detection of fluorescence in situ hybridization of DNA probes and high resolution fluorescent R banding is described. Human lymphocytes were stimulated with phytohemagglutinin and synchronized using a fluorouracil block followed by exposure to bromodeoxyuridine and Hoechst 33258 prior to harvest. Metaphase preparations were treated with Hoechst 33258 and exposed to UV light. Thereafter they were incubated in sodium phosphate buffer and dried prior to in situ hybridization with a biotin-labelled centromere-specific alpha satellite DNA probe for chromosome 1 (pUCl.77) and two digoxigenin-labelled probes, i.e., a PCR-generated chromosome 8-specific alphoid probe (#8) and a cosmid probe for FLT4 gene on 5q33-qter (class III receptor tyrosine kinase). Hybridization signals were detected by an indirect immunofluorescence method using fluorescein isothiocyanate. The chromosomes were counterstained with propidium iodide and 4',6-diamidino-2-phenylindole dihydrochloride. This simple method allows unambiguous chromosome band identification simultaneously with detection of the hybridized probes. PMID- 8244759 TI - On the estimation of heritability for post-emergence longevity in Drosophila melanogaster. PMID- 8244760 TI - Significance of biometrical methodology in our study of longevity genetics. PMID- 8244761 TI - Increased synthesis and specific localization of a major lysosomal membrane sialoglycoprotein (LGP107) at the ruffled border membrane of active osteoclasts. AB - The immunocytochemical localization was investigated of a major lysosomal membrane sialoglycoprotein with a molecular mass of 107 kDa, which was designated as LGP107. The study utilized rat osteoclasts with different bone resorbing activity and osteoclast precursors at various stages of differentiation and maturation together with monospecific antibodies to this protein. Despite its localization primarily in lysosomes and endosomes in the other cell types examined, LGP107 was exclusively confined to the apical plasma membrane at the ruffled border of the active osteoclast, where the osteoclast is in contact with the bone surface. The protein was also concentrated in a number of endocytic vacuoles in the vicinity of the ruffled border membrane. However the labeling was not found in the basolateral membranes of the active osteoclast. The ruffled border membrane detached from the bone surface showed a marked decrease in the extent of the immunolabeling. The post- and/or resting osteoclasts, which were located away from the bone surface, were totally devoid of the membraneous localization of LGP107. No definite immunolabeling was found in the immature preosteoclasts. These results indicate that the protein is largely synthesized in the active osteoclast and rapidly translocated to the ruffled border membrane by vectorial vesicle transport. LGP107 is suggested to contribute to the formation and maintenance of the specialized acidic environment for bone resorption. PMID- 8244762 TI - The acinar heterotopy of phosphoenolpyruvate carboxykinase activity in rat liver under the influence of the oestrous cycle. AB - The acinar localization of phosphoenolpyruvate carboxykinase was investigated in livers from untreated female rats during the 4-day oestrous cycle. The results were correlated with plasma levels of insulin, glucagon and oestrogens. Total activity was highest in dioestrus and lowest in metoestrus. The highest activities were present in the periportal zone and decreased continuously towards the perivenous zone in all four phases of the cycle. This general pattern was modified under the influence of the cycle. The periportal-perivenous gradient was subject to cycle dependent changes, being steepest in proestrus. In addition to oestrogens the concentrations of plasma insulin also varied during the female cycle. Insulin was highest in oestrus and lowest in proestrus. Glucagon showed only minor variations. Oestrogen concentrations increased continuously from low values in oestrus to high values in proestrus. These changes were directly related to the changes in the slope of the activity gradient. PMID- 8244763 TI - Vitamin D3 (soltriol) nuclear receptors in abdominal scent gland and skin of Siberian hamster (Phodopus sungorus) localized by autoradiography and immunohistochemistry. AB - In vivo autoradiography with [3H]1,25-dihydroxycholecalciferol (vitamin D, soltriol) and immunostaining with antibodies to vitamin D receptor were applied to identify specific binding sites in the abdominal scent gland of male Siberian hamster (Phodopus sungorus). Nuclear concentration of radiolabeled hormone and receptor antibodies was observed in the corresponding cell types including basal cells of sebaceous glands, cells of the outer hair sheaths and hair bulbs, and also keratinocytes in the epidermis. Cells of the hair dermal papillae and fibroblasts of the dermis did not show nuclear labeling. There was good correspondence between the autoradiographic and immunohistochemical data. The results indicate the presence of receptors for vitamin D-soltriol and suggest a seasonal regulation of scent gland marking activities by this steroid hormone of sunlight in cooperation with the sex steroid testosterone. PMID- 8244764 TI - Immunocytochemical analysis of phosphatidylinositol-specific phospholipase C in PC12 cells: predominance of the delta isoform during neural differentiation. AB - The rat pheochromocytoma PC12 cell line, which differentiates into sympathetic neurons under nerve growth factor (NGF) treatment, contains at least three phosphoinositidase C (PIC) isozymes, PIC beta, PIC gamma, PIC delta. These isozymes have been previously shown to display a different subcellular localization. To determine whether or not NGF induces changes in the presence and/or distribution of PIC isozymes during PC12 neural differentiation, studies were carried out by means of in situ immunocytochemistry. After NGF administration the proliferative activity was progressively reduced to very low levels, as measured by bromodeoxy Uridine incorporation, and a neuron-like morphology was displayed by almost all cells. In unstimulated PC12 cells, PIC beta was detected in the nucleus whereas PIC delta was only cytoplasmic; PIC gamma was found in both cell compartments. In cells treated with NGF for 3 days, neural processes extended to twice the diameter of the cell body; the gamma isoform was concentrated near the nucleus, while the immunoreactivity of the beta form remained constant and the delta form was increased. After 10 days of treatment with NGF, PIC beta was hardly detectable and PIC gamma immunostaining was considerably decreased. On the contrary, PIC delta progressively increased and, after 14 days of NGF exposure, fully differentiated cells displayed an intense labelling of cell body and neurites. In the same cells, PIC beta and PIC gamma were almost negative. These results suggest that NGF dependent neural differentiation is related to the selective down regulation of PIC beta and gamma and the increase of PIC delta isozyme associated with the decrease of cell proliferation. PMID- 8244765 TI - Protein gene product 9.5 and ubiquitin immunoreactivities in rat epididymis epithelium. AB - A quantitative immunohistochemical study was performed of the distribution of protein gene product 9.5 (PGP, a soluble protein localized in neurons and neuroendocrine cells as well as in some non-nervous cells) and ubiquitin along the rat epididymis. In the ductuli efferents, PGP immunoreaction was observed in the whole cytoplasm of some columnar cells; a smaller number of columnar cells showed ubiquitin immunoreactivity with limited apical and basal cytoplasmic localization. In the proximal caput epididymidis, the whole cytoplasm of all columnar cells showed PGP immunoreactivity, ubiquitin immunostaining was negative in this region. In the middle and distal caput epididymidis and the distal cauda, the apical cytoplasm of some columnar cells and the whole cytoplasm of some basal cells showed immunoreactivity to PGP. In these regions, immunoreactivity to ubiquitin was positive in the supranuclear cytoplasm of some columnar cells but not in the basal cells. No immunoreactivity to PGP or ubiquitin was detected in the corpus epididymis and the proximal cauda. Double immunostaining revealed that all the epididymal ubiquitin immunoreactive cells were also PGP immunoreactive, whereas most PGP immunoreactive cells did not immunoreact to ubiquitin. In ubiquitin-PGP immunoreactive cells, the site of the PGP immunoreaction differed from that of the ubiquitin immunoreaction. PGP-ubiquitin immunoreactive cells also seemed to be immunoreactive to anti-AE1/AE3 keratin antibodies. The spermatozoal heads were immunoreactive to PGP antibodies in the epididymal regions from proximal caput to distal cauda but not in the ductuli efferents. The findings suggest that non-ubiquitinated PGP immunoreactive proteins are secreted in the epididymis, mainly in the proximal caput, and attach to spermatozoa. PMID- 8244766 TI - The continuum of pure and hybrid myosin heavy chain-based fibre types in rat skeletal muscle. AB - The myosin heavy chain (MHC)-based fibre composition of adult rat adductor magnus (AM) and tibialis anterior (TA) muscles was investigated using single fibre analysis. Microelectrophoresis performed on single fibre fragments demonstrated a predominance of pure fast MHC-based fibre types (expressing only one fast MHC). Most of the fibres analysed from both the AM (72%) and TA (50%) were pure type IIB (expressing only MHCIId). Pure type IID fibres (expressing only MHCIId) were also abundant in AM (20%) and TA (18%). In addition, hybrid fibres coexpressing MHCIIb and MHCIId in varying proportions (fibre types IIBD and IIDB) were found, as well as fibres coexpressing MHCIId and MHCIIa with a predominance of MHCIId (type IIDA) and some C fibres (coexpressing MHCI and MHCIIa in varying proportions). Considered altogether, these data reflect the dynamic nature of adult skeletal muscle fibres and indicate a continuum of MHC-based fibre types in normal rat muscle with transitions in the order IIB<==>IIBD<==>IIDB<==>IID<==>IIDA<==>IIAD<==>II A<==>IIC<==>IC<==>I. PMID- 8244767 TI - Demonstration of cells of the mononuclear phagocyte lineage in the periodontium following experimental tooth movement in the rat. An immunohistochemical study using monoclonal antibodies ED1 und ED2 on paraffin-embedded tissues. AB - In this immunohistochemical study two monoclonal antibodies, ED1 and ED2, which recognize exclusively cells of the mononuclear phagocyte system (MPS) in the rat, were applied to study the presence of these cells during remodeling of the periodontal tissues following mechanically induced orthodontic tooth movement. The immunohistochemical procedure was carried out successfully on routinely processed, paraffin-embedded histological sections using the avidin-biotin peroxidase-complex (ABC) technique. Cells of the MPS could be demonstrated on positive control sections of rat spleen and bone marrow. For the study of remodelling of the periodontal tissues only the ED1 antibody proved to be suitable. With this antibody, positive mononuclear and multinuclear cells, i.e. macrophages and osteoclasts, were seen throughout the periodontium even in the control animals. After the induction of orthodontic tooth movement activation of macrophages, osteoclasts and odontoclasts was demonstrable, all of them showing a clear-cut positive reaction to ED1. PMID- 8244768 TI - Confocal images of marrow stromal (Westen-Bainton) cells. AB - A cytochemical method was used for imaging a defined subset of marrow stromal cells (alkaline phosphatase-positive reticulum cells, hereinafter referred to as Westen-Bainton cells), which are endowed with membrane-associated alkaline phosphatase. The use of two different types of confocal microscopes was compared: a tandem scanning reflected light microscope and a laser scanning confocal microscope equipped with a 633 nm (helium-neon) laser. Sharp confocal reflection images of the cytochemically stained stromal cells were obtained with both microscopes. Three-dimensional reconstructions were generated with both systems, revealing morphological features of Westen-Bainton cells related to both their actual shape and organization within tissue architecture, which were not otherwise appreciated. The observations were extended to individual cases of bone pathology, and demonstrated the value of confocal microscopy for the investigation of marrow-bone relationships in physiology and disease. PMID- 8244770 TI - Remodelling of cardiomyocyte cytoarchitecture visualized by three-dimensional (3D) confocal microscopy. AB - The break-down and reassembly of myofibrils in long-term cultures of adult rat cardiomyocytes was investigated by a novel combination of confocal laser scanning microscopy and three-dimensional image reconstruction, referred to as FTCS, to visualize the morphological changes these cells undergo in culture. FTCS is discussed as an alternative imaging mode to low-magnification scanning electron microscopy. The three-dimensional shape of the cells are correlated with the assembly state of myofibrils in different stages. Based on immunofluorescence and confocal laser scanning microscopy it was shown that myofibrils are degraded within a few days after plating and that newly assembled myofibrils are predominantly confined to the continuous area in the perinuclear region close to the membrane in contact with the substratum. The localization of myofibrils along the cell's vertical axis has been investigated both by optical sectioning using confocal light microscopy and by physical sectioning followed by transmission electron microscopy. Based on the distribution of myofibrillar proteins we propose a model of myofibrillar growth locating the putative assembly sites to a region concentric around the nuclei. We provide evidence that the cell shape is dominated by the myofibrillar apparatus. PMID- 8244769 TI - Robert Feulgen Prize Lecture 1993. The journey of the insulin receptor into the cell: from cellular biology to pathophysiology. AB - The data that we have reviewed indicate that insulin binds to a specific cell surface receptor. The complex then becomes involved in a series of steps which lead the insulin-receptor complex to be internalized and rapidly delivered to endosomes. From this sorting station, the hormone is targeted to lysosomes to be degraded while the receptor is recycled back to the cell surface. This sequence of events presents two degrees of ligand specificity: (a) The first step is ligand-dependent and requires insulin-induced receptor phosphorylation of specific tyrosine residues. It consists in the surface redistribution of the receptor from microvilli where it preferentially localizes in its unoccupied form. (b) The second step is more general and consists in the association with clathrin-coated pits which represents the internalization gate common to many receptors. This sequence of events participates in the regulation of the biological action of the hormone and can thus be implicated in the pathophysiology of diabetes mellitus and various extreme insulin resistance syndromes, including type A extreme insulin resistance, leprechaunism, and Rabson Mendehall syndrome. Alterations of the internalization process can result either from intrinsic abnormalities of the receptor or from more general alteration of the plasma membrane or of the cell metabolism. Type I diabetes is an example of the latter possibility, since general impairment of endocytosis could contribute to extracellular matrix accumulation and to an increase in blood cholesterol. Thus, better characterization of the molecular and cellular biology of the insulin receptor and of its journey inside the cell definitely leads to better understanding of disease states, including diabetes. PMID- 8244771 TI - Gamma-aminobutyric acid (GABA) immunoreactivity in the mouse adrenal gland. AB - Gamma-aminobutyric acid (GABA) immunoreactivity was revealed by immunocytochemistry in the mouse adrenal gland at the light and electron microscopic levels. Groups of weakly or faintly GABA immunoreactive chromaffin cells were often seen in the adrenal medulla. By means of immunohistochemistry combined with fluorescent microscopy, these GABA immunoreactive chromaffin cells showed noradrenaline fluorescence. The immunoreaction product was seen mainly in the granular cores of these noradrenaline cells. These results suggest the co existence of GABA and noradrenaline within the chromaffin granules. Sometimes thick or thin bundles of GABA immunoreactive nerve fibers with or without varicosities were found running through the cortex directly into the medulla. In the medulla, GABA immunoreactive varicose nerve fibers were numerous and were often in close contact with small adrenaline cells and large ganglion cells; a few, however, surrounded clusters of the noradrenaline cells, where membrane specializations were formed. Single GABA immunoreactive nerve fibers, and thin or thick bundles of the immunoreactive varicose nerve fibers ran along the blood vessels in the medulla. The immunoreaction deposits were observed diffusely in the axoplasm and in small agranular vesicles of the GABA immunoreactive nerve fibers. Since no ganglion cells with GABA immunoreactivity were found in the adrenal gland, the GABA immunoreactive nerve fibers are regarded as extrinsic in origin. PMID- 8244772 TI - Immunohistochemical localization of xanthine oxidase in human cardiac and skeletal muscle. AB - The generation of a monoclonal antibody specific to xanthine oxidase and its use in the distribution of the enzyme in human tissue is described. Xanthine oxidase was purified from human and bovine milk by a rapid method, allowing for minimal proteolytic degradation, and the purified enzyme preparations were used for the immunization of BALB/c mice as well as for the subsequent selection of hybridomas. The hybridoma clone X1-7, IgG (2a, kappa-light chain) was selected for further analysis and demonstrated to precipitate xanthine oxidase from human liver and skeletal muscle extracts. As determined by SDS-polyacrylamide gel electrophoresis of eluates from affinity chromatography, the X1-7 antibody bound to a main protein of 155 kDa, from human milk and skeletal muscle, and to proteins of 155, 143 and 95 kDa from human liver. Immunohistochemical studies, using two of the monoclonal antibodies with differing epitope specificity, revealed xanthine oxidase to be localized mainly in the vascular smooth muscle cells but also in a proportion of endothelial cells of capillaries and smaller vessels in both human cardiac and skeletal muscle. Immunoreactivity was additionally observed in human macrophages and mast cells. The results of the present study confirm previous reports of the presence of xanthine oxidase in capillary endothelial cells, but also demonstrates additional localization of the enzyme in vascular smooth muscle cells, macrophages and mast cells. The current findings verify that the distribution of xanthine oxidase in human tissue includes cardiac and skeletal muscle. PMID- 8244773 TI - Differential kinetics of propidium iodide uptake in apoptotic and necrotic thymocytes. AB - Apoptosis and necrosis represent two different mechanisms by which cells die. The dynamics of cellular lesions in these two processes differ. In particular we demonstrate that plasma membrane damage, occurring as a primary event during necrosis represents, on the contrary, a delayed but massive phenomenon during apoptosis. In consequence there are different kinetics of propidium iodide incorporation by necrotic and apoptotic thymocytes. This represents the basis for the flow cytometric identification of different cellular subsets. Analysis of these subsets after sorting showed that clearly apoptotic cells, which are not able to exclude propidium iodide for long incubation periods, do not show any morphologically detectable membrane damage. The kinetics of propidium iodide incorporation in vivo in isolated rat thymocytes can therefore be used in flow cytometric analysis. This technique can be used instead of DNA staining of ethanol-treated cells or nick translation to recognize apoptotic cells, and distinguish apoptosis from necrosis, without killing the cell. PMID- 8244774 TI - Ultrastructural organization of type XI collagen in fetal bovine epiphyseal cartilage. AB - Type XI collagen was localized with polyclonal antibodies specific for alpha 1 (XI) and alpha 2 (XI) chains in the resting zone of epiphyseal cartilage from calf fetuses. The immunofluorescence technique was used on sections of cartilage, and the immunogold labelling technique for electron microscopy on fibrils isolated from cartilage and, for the first time, in situ on blocks of cartilage fractured in liquid nitrogen. Immunofluorescence showed that without pepsin treatment the staining of type XI collagen was restricted to the pericellular zones; after pepsin treatment, the staining was co-distributed with that of type II collagen. Immunoelectron microscopy performed on isolated fibrils and on cartilage blocks showed that after disruption of fibrils with pepsin, type XI collagen was labelled on small filaments on the fibrils. When the fibrils were not disrupted, labelling was observed in situ only at the ends of the fibrils or on cross-sections of some fibrils. These results indicate that type XI collagen is located inside type II collagen fibrils in fetal bovine epiphyseal cartilage, as already postulated for embryonic chicken sterna. PMID- 8244775 TI - Predominant expression of the beta subunit of prolyl 4-hydroxylase (disulfide isomerase) in human extravillous trophoblasts. AB - Prolyl 4-hydroxylase is a heterodimeric enzyme that is crucial in the biosynthesis of collagen. The beta subunit of this enzyme is a multifunctional protein which is also known as protein-disulfide isomerase. Immunofluorescence and monoclonal antibody (Mab) 5B5 were used to localize the beta subunit in human extraembryonic tissues. The strongest sites of 5B5 reactivity were extravillous cytotrophoblasts in the basal plate, uteroplacental arteries and amniochorion, syncytiotrophoblast displayed variable weaker reactivity. Only a small fraction of placental 5B5 antigen was detected as a component of prolyl-4-hydroxylase by affinity chromatography on immobilized polyproline. The results indicate a difference in the expression of an endoplasmic reticulum marker between villous and extravillous trophoblast. The predominance of 5B5 antigen in extravillous trophoblast could be associated with an increased ability to synthesize collagen or other enzymatic reactions associated with prolyl 4-hydroxylase beta subunit. PMID- 8244776 TI - Deficit of HLA homozygotes in a Caucasian isolate. AB - Deficits of HLA-A, -B homozygotes observed many years ago in two inbred populations suggested negative selection against HLA homozygotes. To determine whether a similar deficiency would be observed in the S-leut Hutterites, a well characterized Caucasian isolate of European ancestry, and to determine whether selection operated at the allele, locus, or haplotype level, observed and expected numbers of homozygotes were compared in 852 adult Hutterites. Deficits ranging from 11% to 24% were observed for all five loci examined (HLA-A, -B, -C, DR, and -DQ). However, these deficits were secondary to, and almost completely accounted for by, a 64% loss of individuals homozygous for the haplotype. There was no evidence of deficits affecting only a single allele or locus. The data indicate strong negative selection against HLA homozygotes. This could be due, at least in part, to decreased fecundability among couples sharing HLA-DR. However, these data suggest that additional selective factors acting at the level of the haplotype also operate in this population. PMID- 8244777 TI - Lymphokine gene transcription in CD4+CD8- T cells of a type III bare lymphocyte syndrome patient. AB - In this study we analyzed the impact of a MHC class-II-deficient environment on the differentiation of CD4+CD8- T lymphocytes into functional defined subsets of lymphokine-producing T-helper cells. To this end a CD4+CD8- T-cell line and CD4+CD8- T-cell clones, isolated from PBMCs of a type III BLS patient, were stimulated in vitro with anti-CD3 and PMA and assessed for lymphokine transcription patterns. The results of these analyses show that CD4+CD8- T cells that have matured in a MHC class-II-deficient environment display lymphokine transcription patterns that resemble those of MHC class-II-expressing family control-derived CD4+CD8- T cells. PMID- 8244778 TI - Mouse mammary tumor virus-mediated T-cell receptor negative selection in HLA-DRA transgenic mice. AB - Products of specific mouse Mtv genes expressed in association with mouse MHC class II products cause the deletion of T cells expressing particular TCR V beta gene segments. These endogenous deletion ligands have been termed superantigens due to their ability to negatively select entire T-cell families, as defined by V beta-chain usage. In most cases, deletion is preferentially effected through interaction of the Mtv ligand with H-2E products. Although human DR alpha shares only 75% identity with the E alpha chain of H-2E, it has previously been shown to substitute for the mouse homologue in its capacity to induce the deletion of V beta 11- and V beta 17a-bearing T cells. In the present study, we have undertaken a more comprehensive analysis of the interaction of mixed DR alpha/E beta pairs with various endogenous Mtv integrants in various mouse backgrounds, leading to negative selection of particular V beta families. We show in this paper that transgenic DR alpha/E beta can also efficiently interact with products of Mtv-7, causing deletion of both V beta 6+ and V beta 7+ cells. Deletion of V beta 11+ T cells in DRA transgenic mice carrying Mtv-8 and -9, however, was less efficient than in control H-2Ea transgenic mice. These data and those from other MHC transgenic mouse studies show that while the class II alpha chain can influence the interaction with superantigen, it is the identity of the beta chain that seems to be critical.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244779 TI - Lack of association of HLA polymorphisms with human papillomavirus-related cervical cancer. AB - An association of HLA-DQ3 with SCC of the cervix has been reported by researchers in Germany and Norway. This article documents a similar-sized study with patients and controls from northwest England. We report in detail on serologically determined HLA polymorphism in SCC patients with respect to HPV 16 infection, MHC class II expression within the tumor, serologic response to HPV, and other relevant clinical variables. We have also extended our studies to include DNA based analysis using PCR and SSO probes for HLA-DQ. No significant association of any HLA-A, -B, -C, -DR, or -DQ antigen with SCC patients was found. While a possible explanation of the differences among studies could be a reflection of disease heterogeneity, the several tumor and clinical factors examined do not account for the observed differences from previous reports. Further studies are needed for a greater understanding of the interaction of HPV and HLA type in the development of cervical neoplasia. PMID- 8244780 TI - Restricted usage of T-cell receptor V alpha sequence and variable-joining pairs after normal T-cell development and bone marrow transplantation. AB - TCR V alpha 3 and V alpha 5 transcripts in PBLs from healthy individuals of multiple age groups and from BMT recipients were analyzed. PCR, cloning, and sequencing studies revealed significant V-J junctional diversity among TCR transcripts from all tested blood samples, as provided both by N/P-region addition and exonuclease activity. However, results illustrated restrictions in TCR alpha diversity at several additional levels. First, V alpha 5 and V alpha 3 gene families, which were expected to be composed of multiple members, were dominated in each case by a single sequence at the transcript level. Second, restrictions existed in V-J pairing in that J alpha genes, which were encoded toward the 5' region of the locus, were rearranged frequently with V alpha 3 and rarely with V alpha 5. Conversely, J alpha genes encoded toward the 3' region of the locus preferentially rearranged with V alpha 5. Healthy individuals showed few differences with regard to V-J pairing patterns, while one of three BMT recipients demonstrated a skewed usage of 3' J alpha genes. In total, results demonstrated qualitative restrictions that may limit the working TCR repertoire in human peripheral tissues, both among BMT recipients and their healthy donors. PMID- 8244781 TI - The influence of HLA-DR and -DQ alleles on progression to multiple sclerosis following a clinically isolated syndrome. AB - A 5-year follow-up study was performed on 70 Caucasian patients presenting with isolated neurological syndromes of the optic nerve, brain stem, or spinal cord to assess the risk of progression to MS. The influence on patient prognosis of HLA DR and -DQ alleles and presentation with disseminated brain lesions, demonstrated by MRI scanning, was determined. Clinical progression to MS was observed in 61% of optic neuritis patients, 50% of patients with a brain-stem syndrome, and 35% of patients with a spinal cord disturbance. MS and the isolated clinical syndromes were positively associated with DRB1*1501, DQA1*0102, and DQB1*0602; the frequency of these alleles in the latter group was intermediate between that seen in MS patients and healthy controls. Conversion to MS was positively associated with the DRB1*1501.DQA1*0102.DQB1*0602 haplotype, but the influence of HLA was only significant in patients with disseminated brain lesions at presentation (MRI positive); MS developed in 86% of MRI-positive, DRB1*1501 positive patients compared with 55% of MRI-positive, DRB1*1501-negative patients (p < 0.025). The data suggest that these HLA alleles are involved in susceptibility to initial demyelinating lesion formation and are important in the subsequent development of MS in MRI-positive patients. PMID- 8244782 TI - Molecular characterization of HLA-B71 from an African American individual. AB - We have sequenced a cDNA clone encoding an HLA-B71 (B70) allele from an African American individual. Serologic definition of B70 allelic products is very difficult due to extensive cross-reactivity of the alloantisera with B35, B15, and B5 antigens. The new sequence most closely resembles the sequence of B*1503, differing only by three amino acids at positions 63, 67, and 116. The B71 sequence differs from alleles of the B15 antigenic group (serologically defined as B62) by 7-8 amino acids and from members of the B35 family by 10 to 12 amino acids. B71 may represent an evolutionary intermediate, sharing elements common to both B35 and B15 allelic groups. PMID- 8244783 TI - Nucleotide sequence of a novel DPB1 allele, DPB1*4701. PMID- 8244784 TI - 'Osteopathic physician' defines our identity. PMID- 8244785 TI - OMT enhances any medical regimen. PMID- 8244786 TI - Primary care DOs, specialists--and OMM--important to profession. PMID- 8244787 TI - Quantifying passive resistance to motion in the straight-leg-raising test on asymptomatic subjects. AB - The clinician needs an objective way to measure limb motion in the straight-leg raising test. A biomechanical algorithm was used to quantify resistance to motion in 15 asymptomatic subjects. Measurements from a pendulum electrogoniometer and hand-held load cell were used to calculate a moment representing passive resistance to motion. After three measurement trials, significant increases in range of motion (4.7%) and moment (8.4%) occurred. Then, an isometric contraction relaxation of the hip extensors produced a highly significant increase in motion (8.8%) but decrease in moment (< 14.3%). A third order polynomial fit of moment per angle stratified the sample into two groups according to their change in moment. Motion in group 1 increased 8.0%, and in group 2, 9.5%. However, group 1 had no change in moment whereas group 2 had a highly significant decrease in moment (22.9%). The measured change in resistance demonstrated that a simple biomechanical algorithm quantified properties in a clinical test that were not observed in range of motion alone. PMID- 8244788 TI - Delirium tremens: a review. AB - Delirium tremens was first identified as being due to long-term excessive alcohol intake in 1813, but is now known to be associated with abrupt withdrawal of alcohol in chronically habituated persons. Recent publications quote an anticipated mortality rate of 15% to 20%. Our experience in the past 20 years has not confirmed that rate. This review reveals that the prevalence of fatal cases is extremely low, with the true mortality close to 0%. We believe that this decrement is due to the increasing use of benzodiazepines to detoxify alcoholic patients. It is postulated that the benzodiazepines act either to prevent delirium tremens or to reduce the neurotransmitter disruption in the central nervous system caused by excessive alcohol intake, or both. PMID- 8244789 TI - Teaching residents to write a research paper. AB - Medical writing and publications are important in developing a scholarly basis for residency programs and in providing a learning experience for both resident and faculty mentors. Residency directors must provide the stimulus and support for both faculty and residents' varied creative activities. This support manifests itself in a commitment to scholarly activity (including a dedicated research person), the procurement of available research materials, the establishment of a process or plan for beginning a research project, and the development of a method for rewarding or recognizing faculty and residents who produce scholarly works. Some osteopathic residency programs may need to train faculty in research skills at the same time that residents are learning to write. Trained faculty are better models and coaches for residents engaged in research. Beginning with a fundamental, but disciplined, writing program, both faculty and residents may learn methods for sharing new knowledge or acquiring those skills necessary to critically analyze the medical literature. PMID- 8244790 TI - Complications of varicella pneumonia in adults. AB - Varicella is predominantly a disease of children, in whom it typically has a benign course and outcome. However, when the disease affects adults, the complications can be life-threatening. Varicella pneumonia, secondary bacterial infection, and opportunistic infection in the immunocompromised host are the most common complications in the adult. Surveillance for these complications is mandatory because appropriate therapy may be life-saving. The introduction of specific antiviral agents may be useful in certain clinical circumstances. The authors describe two representative patients and discuss the diagnosis and treatment. They also discuss this infection in adulthood in general. PMID- 8244791 TI - Why prevent child maltreatment? PMID- 8244792 TI - State of the art of nursing research and theory development in adolescent health. AB - A literature review was conducted to ascertain the state of the art of nursing research and theory development in adolescent health. A conceptual framework and data collection tool were developed to guide evaluation of the research reports. Adolescent health literature reported within the past 10 years (June 1982 to June 1992) was reviewed. Of 124 adolescent health articles identified in literature searches, 46.0% (n = 57) met inclusion criteria and were included in the literature review. A developmental approach, which included the purposes of theory and research (Chinn & Jacobs, 1987), was used to determine the state of the art of nursing research and theory development. Nursing research and theory development in adolescent health were determined to be at a level between adolescence and young adulthood, reflecting beginning maturity. PMID- 8244793 TI - Home care for medically fragile children: urban versus rural settings. AB - The development of managed care programs has allowed children who are medically fragile and technology-dependent to be discharged back to their home communities. Rural and urban settings each have unique characteristics that can affect home care. A number of home-care problems are common to both urban and rural settings, including nursing coverage, respite care, school services, coordination of medical resources, and emergency response protocols. Although the problems may be similar, the solutions often differ and should be adapted to the unique urban or rural settings in which they occur. Approaches to these problems are offered on the basis of experience in providing care management to these children in the Specialized Home Care Program in Michigan. Also explored are the problems of the lack of resources available to children who are medically fragile but not technology-dependent. PMID- 8244794 TI - Crucial issues in the treatment of a sexually abused latency-aged boy. PMID- 8244795 TI - The relationship between maternal characteristics, maternal vulnerability beliefs, and immunization compliance. AB - The purpose of this descriptive study was to determine the relationship of demographic variables and mothers' perceived vulnerability to eight communicable diseases with immunization compliance. A random sample consisting of 40 mothers with preschool-age children 4-24 months of age was interviewed by means of the Communicable Disease Perceived Vulnerability Scale and the investigator-developed Demographic Data Collection Instrument. Data were analyzed with descriptive statistics and chi-square (p < or = .05). The average mother in the sample was White, had a 10th- to 12th-grade education, was 24-31 years old, and had a yearly income of $10,000-$45,000. Statistically there were no significant relationships between maternal characteristics or between maternal vulnerability beliefs and immunization compliance. However, it is clinically significant that the overall compliance rate of 70% in this rural area is much higher than the national average of 50%. Findings suggest that using health education materials at the time of newborn discharge, mailing reminder cards, and accessing a statewide computer recall system may increase immunization compliance. In addition, mothers who perceived a vulnerability to the diseases had a 60.6% compliance rate, suggesting that perception of vulnerability may be a useful predictor of immunization compliance. PMID- 8244796 TI - Health needs of school-age children in two midwestern counties. AB - Administrators, school nurses, and informed parents (N = 113) were queried by means of the Delphi Technique to determine priority health needs and barriers in two midwestern counties. Needs of health education and school nurses and barriers of restricted resources received top ranking by all participant groups. Descriptive measures were computed by using Kendall's tau. PMID- 8244797 TI - Billboards in space: a health care opportunity? PMID- 8244798 TI - Parental perceptions and support strategies in caring for a child with a chronic condition. AB - The purpose of this article is to compare fathers' and mothers' perceptions of a child's chronic condition and to compare strategies used by fathers and mothers to support themselves during critical times. Analysis of data focused on questionnaires from 38 fathers and mothers (19 paired parents) who had children with chronic conditions. Results showed that, although parental perceptions of the child's chronic condition were similar, their responses to the situation were not the same. Fathers and mothers relied on different strategies to support themselves during critical times. PMID- 8244799 TI - Mothers of chronically ill children and perceptions of environmental variables. AB - Environment plays a critical role in supporting families, but the influence of the environment on families with a chronically ill child has remained relatively unexplored. This paper presents a study that examined the role of the environment in families with a chronically ill child. This study, using a secondary analysis of two existing data bases (Holaday & Turner-Henson, 1991; U.S. Bureau of the Census, 1980), examined the influence of selected environmental factors on mothers' perceptions of the environment as supportive, resourceful, safe, and accessible. A presentation of the study findings, as well as implications for nursing practice, is presented. PMID- 8244800 TI - Day care for ill children: an employed mother's dilemma. AB - The purpose of this qualitative study was to describe the process that employed mothers used to make child care decisions for children who became ill while enrolled in day care and the factors that influenced those decisions. Responding to an investigator-developed interview schedule, 20 married, employed mothers of preschoolers in day care each described a recent experience when her child was ill on a work day, her feelings, advantages and disadvantages of available options, preferences, job characteristics, and demographics. Inductive analysis of narrative data revealed themes of anxiety and decisional conflict between work and motherhood responsibilities and limited care options. These mothers usually cared for their sick children, sometimes sharing the duty with another caregiver. Data were categorized by actually selected and usually selected modes of care: mother; mother and father; mother and grandparent; mother and sitter; and mother and day care center. Influencing factors included: certainty and severity of illness, relative advantages and disadvantages of available options, job flexibility, and availability of paid leave. A model describing the decision making process is proposed. While further research is needed, this model provides some guidance for nursing practice with families who face this dilemma and for evaluation of workplace policies. PMID- 8244801 TI - Comparison of dependent-care activities for well siblings of children with cystic fibrosis and well siblings in families without children with chronic illness. AB - The purpose of this study was to determine if well siblings in families with a child with cystic fibrosis receive less dependent care, as defined by Orem, than well siblings in families without children with chronic illness. Nineteen mothers of children with cystic fibrosis and a well sibling and 19 mothers of only well children completed the Dependent Care Agent Instrument. In this population, no significant differences were found in a mother's performance of dependent-care activities for well siblings of children with cystic fibrosis and well siblings in families without children with a chronic illness. The possibility of a difference in a child's and mother's perception of parenting and the use of a questionnaire to study parenting is discussed. The implication that the child's perception of dependent care received is overlooked by focusing only on the mother's perception warrants further consideration. PMID- 8244802 TI - Homeless children and families: clinical and research issues. AB - The rapid increase in the number of homeless children and families has focused attention on understanding the health and related needs of this vulnerable group. This paper describes challenges associated with providing health care to homeless children and evaluation of those services. Data are presented from the Nursing Center for the Homeless of the School of Nursing at the State University of New York at Buffalo. Health services provided to homeless children and families (N = 470) documented that more than half (56%) were covered by health insurance and had received age-appropriate preventive health care. Of the children seen, 50% were considered well and 30% were diagnosed as having upper respiratory infections, skin problems, and/or gastrointestinal disorders. Health teaching for parents was the most frequent nursing intervention (50%), while 20% of the children were referred to community agencies. Factors that impede data collection and provision of health services for the homeless population are discussed, including suggestions for treatment and research approaches. PMID- 8244803 TI - A selective approach to adjunctive therapy for cancer of the rectum. AB - PURPOSE: To present results of a selective approach to adjunctive therapy and surgery based on a new model of clinical staging for rectal cancer. METHODS AND MATERIALS: Three hundred and sixty-two patients with rectal cancer treated with adjunctive radiation therapy and surgery have been analyzed to define patient selection criteria based on clinical assessment of disease. Clinical prognostic features of tumor mobility and level of lesion in the rectum with reference to the anorectal junction were used. Mobile, early fixed (partial), advanced fixed (total) and frozen pelvis are defined as clinical Stages I, II, III, and IV. Tumors above 6 cm (middle valve), 3-6 cm (inferior to middle valve), 0-3 cm (anorectum to inferior valve), and into the anal canal are defined as levels a, b, c, d, respectively. Based on this model, patients with mobile tumors of the proximal rectum (CS Ia, b) are treated with 500 cGy preoperative radiation and selective postoperative radiation (4500 cGy) for Stages B2 and C cancer. All other patients are treated with escalating doses of preoperative radiation. Follow-up in these patients ranges from 1 year to 14 years with a median of 5 years. RESULTS: Overall 5-year survival of the total group of patients is 69%. Survival by pathological stage is 82% for O, A, B1, 67% for B2, 74% for C1, and 51% for C2. Survival by clinical stages is 77% for CS I, 67% for CS II, 57% for CS III, and 21% for CS IV. Overall local recurrence is 43/362 (12%). L.R. by pathological stages is 5% for O, A, B1, 18% for B2, 10% for C1, and 17% for C2. L.R. by clinical stages is 9% for CS I, 14% for CS II, 17% for CS III, and 50% for CS IV. CONCLUSION: An integrated adjunctive therapy and selective surgical approach based on careful clinical staging of rectal cancer results in a global improvement in overall local control and survival of patients. PMID- 8244804 TI - High-dose preoperative radiation for cancer of the rectum: impact of radiation dose on patterns of failure and survival. AB - PURPOSE: A variety of dose-time schedules are currently used for preoperative radiation therapy of rectal cancer. An analysis of patients treated with high dose preoperative radiation therapy was undertaken to determine the influence of radiation dose on the patterns of failure, survival, and complications. METHODS AND MATERIALS: Two hundred seventy-five patients with localized rectal cancer were treated with high-dose preoperative radiation therapy. One hundred fifty-six patients received 45 Gy, +/- 10% (low-dose group). Since 1985, 119 patients with clinically unfavorable cancers (fixed, low-lying, or deeply ulcerated) were given a higher dose, 55 Gy, +/- 10%, using a shrinking field technique (high-dose group). All patients underwent curative resection. Median follow-up was 66 months in the low-dose group and 28 months in the high-dose group. Patterns of failure, survival, and complications were analyzed as a function of radiation dose. RESULTS: Fourteen percent (38/275) of the total group developed a local recurrence; 20% (31/156) in the low-dose group as compared with 6% (7/119) in the high-dose group. The actuarial local recurrence rate at 5 years was 20% for the low-dose group and 8% for the high-dose group, and approached statistical significance with p = .057. For tethered/fixed tumors the actuarial local recurrence rates at 5 years were 28% and 9%, respectively, with p = .05. Similarly, for low-lying tumors (less than 6 cm from the anorectal junction) the rates were 24% and 9%, respectively, with p = .04. The actuarial rate of distant metastasis was 28% in the low-dose group and 20% in the high-dose group and was not significantly different. Overall actuarial 5-year survival for the total group of patients was 66%. No significant difference in survival was observed between the two groups, despite the higher proportion of unfavorable cancers in the high-dose group. The incidence of complications was 2%, equally distributed between the two groups. CONCLUSION: High-dose preoperative radiation therapy for rectal cancer results in excellent local control rates. However, in clinically unfavorable cancers a higher dose (55 Gy) of preoperative radiation can be given safely with significantly improved local control. Careful clinical staging is helpful in selectively treating patients with unfavorable tumors to a higher preoperative radiation dose and thus optimizing their outcome. PMID- 8244805 TI - The reality of radical sphincter preservation surgery for cancer of the distal 3 cm of rectum following high-dose radiation. AB - PURPOSE: The inordinately high rate of locoregional recurrence following sphincter-preserving surgery for cancer of the distal rectum led to the conviction that restorative surgery was inappropriate for the low level cancer. A rectal cancer management program initiated in 1976 that selectively uses high dose preoperative radiation and sphincter-preserving surgery produced lower than expected local recurrence rates. Exploring the safety of extending the indication for sphincter-sparing surgery to include post-radiation mobile cancers as low as the 0.5 cm level is the purpose of this report. METHODS AND MATERIALS: Of 218 rectal cancer patients treated with high-dose preoperative radiation and sphincter-preserving procedures, 69 had radical curative surgery for cancers at or below the 3 cm level. Data regarding the first 52 patients whose ages ranged from 39 to 77 years form the basis of this report. Fifty-seven percent were men. Twenty-five (48%) patients had post-radiation unfavorable cancers (B2, C1, C2). Forty-five to sixty Gy high energy photon radiation was administered over 4 1/2 to 6 weeks followed by a similar interval prior to radical proctosigmoidectomy with anastomosis in the distal 1 cm of rectum. Temporary fecal diversion was performed in all patients; colostomies were closed after 8 weeks. RESULTS: There was zero mortality and two self-limiting anastomotic leaks. Local recurrence developed in 6/43 (14%) patients followed for 24 months or longer. By stage, there were 0/21 (0%) recurrences among O, A, B1 tumors; 6/22 (27%) among unfavorable tumors. By distal margins 1/9 (11%) occurred in .3-1 cm; 4/13 (31%) 1.1-2 cm; 1/18 (5%) 2.1-3 cm. Five-year Kaplan Meier actuarial survival for the 52 patients was 85%. CONCLUSION: Our data indicates that sphincter preservation can be accomplished in cancers of the distal 3 cm of rectum if high-dose preoperative radiation is administered and fixed cancers are excluded. This is the first reported study of sphincter-preserving surgery for the distal rectal cancer after high-dose radiation. The data are important to the design of new treatment options. PMID- 8244806 TI - Postoperative local radiotherapy in rectal cancer: treatment results with limited radiation fields. AB - PURPOSE: The purpose of this study is to determine the treatment results and complication rates of postoperative local radiotherapy, with doses per fraction of 2.25 to 2.50 Gy, in patients with rectal carcinoma who have received macroscopically radical surgery. METHODS AND MATERIALS: A retrospective analysis was done of the records of all consecutive patients (N = 147) with rectal carcinoma Dukes' Stage B or C who have received postoperative local radiotherapy in our institution in the years 1981 through 1989. All patients have been treated on a limited target area covered by only local radiation fields. Locoregional fields covering the whole iliac node chains have not been given. In our treatment protocol doses per fraction were from 2.25 to 2.50 Gy and total doses from 50 to 55 Gy. The minimum follow-up was 24 months; eight patients have been lost to follow-up. RESULTS: The overall 5-year survival rate for the whole group of patients was 39%. The actuarial 2- and 5-year pelvic recurrence rates were 14% and 22% respectively for Dukes' B patients and 30% and 38% respectively for Dukes' C patients. The difference between the pelvic recurrence rates of Stage B and Stage C patients was statistically significant (p = 0.009). No other factors with prognostic significance for pelvic recurrence were found. The interval between surgery and radiotherapy especially had no influence on pelvic recurrence rates. The 35 pelvic recurrences were classified as follows: 17 in-field, 5 marginal, 1 out-of-field, and 9 peritoneal seeding; in three patients there was not enough information for classification. Of the 32 classified pelvic recurrences, the five marginal recurrences were probably geographical misses, only the one out-of-field recurrences, the five marginal recurrences were probably geographical misses; only the one out-of-field recurrence might have been prevented with locoregional radiotherapy. Serious complications caused by the radiotherapy have occurred in 3% of the patients. CONCLUSION: We conclude that the results of postoperative local radiotherapy alone are comparable with the published results of locoregional radiation. Even when relatively high doses per fraction are given low complication rates are seen. PMID- 8244807 TI - Carcinoma of the cervix: predictive value of clinical and magnetic resonance (MR) imaging assessment of prognostic factors. AB - PURPOSE: This retrospective study assesses the predictive value of magnetic resonance imaging (MRI) to identify high risk cervical cancer patients. METHODS AND MATERIALS: The MRI evaluation of morphologic risk factors in patients with invasive cervical carcinoma treated with definitive radiation therapy were correlated with clinical factors and with complete tumor regression (CTR) at 6 months, tumor local control (TLC), and patient outcome at 12 months after irradiation. Sixty-six patients, median age 44.5 years, with bulky Stage I or greater disease were included in the study. RESULTS: In univariate analysis, clinical International Federation of Gynecology and Obstetrics (FIGO) stage had significant correlation with patient outcome, but it correlated poorly with complete tumor regression and tumor local control. In contrast, MRI stage showed significant correlation with complete tumor regression, tumor local control, and disease-free survival at 12 months. When each stage was analyzed separately, the greatest difference was demonstrated between clinical and MRI assignment of stage Ib disease. MRI Stage Ib disease significantly correlated with all three categories analyzed, while clinical Stage Ib did not. Superiority of MRI assessment of low stage disease was also evident in the detection of lymph node metastasis. Significant risk for nodal metastasis was related to tumor size greater than 4 cm, invasion of the parametria and urinary bladder, and stage of the disease. CONCLUSION: The multivariate analysis demonstrated that the most related variables in order of significance were the presence of juxta-regional and paraaortic lymph nodes, patient age, tumor size, and MRI tumor stage. This study demonstrates the value of MR imaging as an adjunct to clinical assessment of bulky invasive cervical cancer, rendering more complete assessment of morphologic risk factors important in patient prognosis. PMID- 8244808 TI - Treatment results of high dose rate brachytherapy in patients with carcinoma of the cervix. AB - PURPOSE: The combination of external beam irradiation and low-dose-rate brachytherapy is known to be an effective form of treatment in carcinoma of the cervix and any change from this well-established therapeutic combination must be able to equal or improve the treatment results. Since 1984 we have been using high dose rate brachytherapy in conjunction with external beam irradiation for patients with carcinoma of the cervix. This paper reports our long term treatment results in terms of local disease control, survival, and complications. METHODS AND MATERIALS: Between January 1984 and December 1989, 187 previously untreated patients with carcinoma of the cervix underwent combined external beam irradiation and high dose rate brachytherapy. The International Federation of Gynecology and Obstetrics stage distribution of patients was as follows: I B = 15, II A = 35, II B = 68, III A = 9, III B = 54, IV A = 6. External beam irradiation to the whole pelvis was delivered by megavoltage irradiation with once-a-day fractionation, to a median dose of 4600 cGy. High dose rate brachytherapy was delivered by a high-dose-rate remote controlled afterloading unit, containing 20 spherical Cobalt 60 sources with a nominal activity of 19 GBq (0.5 Ci) at the time of installation, giving a typical dose rate to point A of 160 cGy/min, decreasing to about 80 cGy/min at the end of the 5-year study. One to 3 high dose rate brachytherapy treatments delivering 800 to 1000 cGy to point A were given weekly concurrently with the last 2 to 3 weeks of radiation therapy, or following its completion. Maximum rectal and bladder doses were routinely measured for each treatment. RESULTS: Overall 5-year actuarial survivals were as follows: I B = 72%, II A = 65%, II B = 66%, III A = 66%, III B = 45%. Five-year actuarial pelvic control rates were as follows: I B = 66%, II A = 83%, II B = 78%, III A = 88%, III B = 40%. At a median follow-up time of 54 months for patients at risk, 23 patients developed 25 complications attributable to radiotherapy (13 rectal, 3 bladder, 8 small bowel, 1 fistula) at a median time of 18 months following completion of treatment. Thirteen complications (7.6%) were grades 3 or 4. Patients with Stage II disease had a higher incidence of complications than patients with Stages I and III disease (p < 0.05). Rectal complications were significantly higher in patients who received a total rectal dose > 5400 cGy (p = 0.045). CONCLUSION: High-dose-rate brachytherapy treatment results are comparable to those obtained with low dose rate brachytherapy techniques. The use of three high dose rate brachytherapy insertions is a practical, economical, and safe treatment for patients with carcinoma of the cervix. PMID- 8244809 TI - Local control prediction by histologic grading or radiation effects with drill biopsy specimens for uterine cervical cancer. AB - PURPOSE: Local control predictability by histologic grading for radiation effects was analyzed on 224 patients with cervical squamous cell carcinomas obtaining complete regression. METHODS AND MATERIALS: The cervical specimens after radiation therapy were obtained by drill biopsy technic. Radiation effect was assessed with Shimosato's histologic classification for radiation effect. RESULTS: The cancer tissues were altered by radiation to various degree from destruction of cancer nests with numerous viable cancer cells indicating grade 2a to scar tissues without viable or non-viable cancer cells indicating grade 4 at the end of treatment. Forty five percent of the patients had viable cancer cells remaining on the specimens at that time. The numbers of the grades 2a, 2b, 3, and 4 in biopsy specimens were 36, 65, 39, and 84, respectively. The local control rates and 5-year survival rates were 53% and 28% for grades 2a, 75% and 51% for grade 2b, 95% and 74% for grade 3, and 92% and 76% for grade 4, respectively. The histologic grading was significantly associated with the local control and 5-year survival. Of 41 patients having remaining viable cancer cells, residual tumors disappeared in 33 patients (80%) and 8 (20%) still persisted to show grades 2a-b at 1 to 2 months after radiotherapy. Of the 8, all four patients without any salvage treatment, died of local recurrence. CONCLUSION: These results suggested that the remaining viable cancer cells in the specimens at 1 to 2 months after radiation therapy were highly associated with recurrence and that the histologic grading system was an indicator predictive for prognosis. PMID- 8244810 TI - Treatment of endometrial carcinoma with radiation therapy alone. AB - PURPOSE: To review the results of treatment with radiotherapy alone in 152 patients with adenocarcinoma of the endometrium who had medical or surgical contraindications to hysterectomy. METHODS AND MATERIALS: We reviewed the records of all patients who were treated with radiotherapy alone for uterine carcinoma at The University of Texas M. D. Anderson Cancer Center between 1960 and 1986. One hundred fifty-two cases were analyzed. Most patients had multiple medical problems. One hundred sixteen patients were treated with intracavitary radiotherapy alone. A combination of external beam and intracavitary radiotherapy was used for 10 patients with Stage I disease who had unusually large cavities, 10 patients with Stage II disease, and 13 of 15 patients with Stage III or IV disease. Histologic material was reviewed in 91 cases. RESULTS: Ten years after treatment, these patients were twice as likely to have died of intercurrent illness as of uterine cancer. The 5-year disease-specific survival rate of patients with Stage I disease was 87%. The disease-specific survival of patients with Stage II disease was 88%, which was not significantly different from that of Stage I patients. Stage III and IV patients had a significantly poorer disease specific survival rate of 49% at 5 years. Intrauterine recurrence occurred in 14% of the patients with Stage I or II disease. Salvage treatment was attempted in 5 of the 10 patients who had isolated intrauterine recurrences of Stage I disease and was successful in all cases. Extrauterine pelvic recurrence developed in only 3% of Stage I and II patients. Of 82 Stage I and II carcinomas that were available for pathologic review, 17 (21%) were clear-cell or papillary serous variants. The disease-specific survival rate of patients with Stage I or II papillary serous carcinomas was 43%, significantly poorer than that of patients with endometrioid carcinomas. Seven patients experienced acute anesthesia-related complications; none were fatal. Five patients had serious late complications of radiation therapy. CONCLUSION: Radical radiotherapy achieved acceptable DSS and local control rates in patients with medically or surgically inoperable uterine carcinoma. However for patients with localized disease, such treatment is justified only when the operative risk exceeds the 10-15% uterine recurrence rate expected with radiation alone. PMID- 8244811 TI - Vaginal stenosis and sexual function following intracavitary radiation for the treatment of cervical and endometrial carcinoma. AB - PURPOSE: This study was designed to document the incidence and degree of vaginal stenosis, sexual activity, and satisfaction in women treated with intracavitary radiation therapy for cervical or endometrial carcinoma. METHODS AND MATERIALS: The incidence of vaginal stenosis in 90 patients treated for either cervical carcinoma (n = 42) or endometrial carcinoma (n = 48) with standard doses of intracavitary radiation from 1989 to 1992 with at least 6 months follow-up was recorded. Vaginal length was measured using a modified vaginal dilator calibrated in centimeters. Semistructured patient interviews documented pretreatment and posttreatment sexual activity and sexual satisfaction. RESULTS: Vaginal length was found to be dependent on diagnosis (cervical vs. endometrial), stage (I vs. II/III), and follow-up interval (6-12 months, 12-24 months, and > 24 months). Slightly more women reported sexual activity post vs. pretreatment (31% active pretreatment vs. 43% active posttreatment). However, 22% of women reported a decrease in sexual frequency and 37% reported a decrease in sexual satisfaction. Reports for all parameters were worse for women treated for cervical carcinoma vs. women treated for endometrial carcinoma with the exception of dyspareunia. CONCLUSION: High-dose radiation for either cervical or endometrial carcinoma at standard doses with or without hysterectomy can cause a decrease in vaginal length as compared to the normal vaginal length of 8 to 9 cm documented by Masters and Johnson. Women treated with intracavitary implants remained as sexually active postimplant as preimplant, however, coital frequency and sexual satisfaction decreased while dyspareunia increased. The decrease in vaginal length may explain in part the decrease in sexual satisfaction but cause and effect have not been established. PMID- 8244812 TI - Patterns of disease recurrence following definitive therapy of adenocarcinoma of the pancreas using surgery and adjuvant radiotherapy:correlations of a clinical trial. AB - PURPOSE: Between 1980 and 1984, 26 patients with resectable adenocarcinoma of the pancreatic head were enrolled in a National Cancer Institute protocol evaluating intraoperative radiotherapy vs. standard therapy. METHODS AND MATERIALS: After complete excision of their lesions, patients were observed (Stage I), or randomized to intraoperative radiotherapy vs. external beam radiotherapy (Stages II-IV). The intraoperative dose was 20 Gy in a single fraction using 9-20 MeV electrons. The external beam radiotherapy schema involved daily 150-180 cGy fractions to 45-55 Gy in 5-6 weeks. Chemotherapy was not used for primary disease but was administered off protocol for recurrent disease. Median potential followup for the trial was > 9 years, with a median patient survival of 18 months. Perioperative mortality was 27% (7 patients). Of the remaining 19 patients, one remains alive and without evidence of disease 9 years post-therapy. Twelve patients underwent autopsy and 2 required antemortem laparotomy; histopathologic evidence of disease recurrence was analyzed. RESULTS: Of 15 patients evaluable for intra-abdominal control, 7 (47%) suffered local recurrences and 7 (47%) failed regionally, with 5 patients (35%) failing in both areas. Five patients (35%) developed peritoneal seeding. Of 13 patients evaluable for systemic disease, 8 (62%) suffered distant failure. There were no differences in outcome between intraoperative or external beam radiotherapy or observation in this subset of patients. CONCLUSION: This report is among the most rigorous descriptions of patterns of failure after resection of pancreatic cancer. It appears clear that advances in local control of this disease are unlikely to translate into increased survival in the absence of improved systemic therapy. PMID- 8244813 TI - Esthesioneuroblastoma: the role of adjuvant radiation therapy. AB - PURPOSE: We describe our experience with adjuvant radiation therapy in patients who underwent operation for esthesioneuroblastoma. METHODS AND MATERIALS: Between January 1951 and December 1990, 49 patients with esthesioneuroblastoma received their initial treatment at the Mayo Clinic. There were 27 male and 22 female patients; their ages ranged from 3 to 79 years (median, 54 years). The tumors were Kadish Stage A in 4 patients, Stage B in 13, Stage C in 29, and modified Kadish Stage D in three (cervical nodal or distant metastasis). The tumors were graded according to Hyams' classification. Treatment included gross total resection alone in 22 patients and gross total resection and postoperative adjuvant radiation therapy in 16. The patients treated with adjuvant radiation had a greater proportion of advanced-stage and high-grade tumors. RESULTS: The 5 year actuarial overall survival, disease-free survival, and local control rates were 69.1% + 7.0%, 54.8% + 7.6%, and 65.3% + 7.4%, respectively. The only significant predictor for overall survival, disease-free survival, and local control was Hyams' grade. Local control was improved in patients who received postoperative adjuvant radiation even though this group of patients had more advanced and higher-grade tumors (5-year rate of local control was 85.9% + 9.3%, compared with 72.7% + 9.5% for those who had operation alone, p = 0.26). CONCLUSION: Adjuvant radiation therapy for esthesioneuroblastoma improves local tumor control, particularly for high-grade and high-stage tumors. We recommend additional treatment with radiation (55.5 Gy) after complete resection of esthesioneuroblastoma. PMID- 8244814 TI - Palladium-103 versus iodine-125 for ophthalmic plaque radiotherapy. AB - PURPOSE: A dosimetry study comparing the use of I-125 vs. Pd-103 radioactive seeds for ophthalmic plaque brachytherapy. METHODS AND MATERIALS: Palladium-103 (Pd-103) seeds in ophthalmic plaques were used to treat 15 patients with intraocular malignant melanoma. Computer-aided simulations were performed to evaluate the intraocular dose distribution of I-125 versus Pd-103 ophthalmic plaques (delivering equivalent apex doses). Seven target points were selected. Starting at the outer scleral surface, four were located along the central axis of the plaque: the 1 mm point (the inner sclera), the 6 mm point, the tumors apex, and the opposite eye wall. We also evaluated the fovea, optic nerve, and the lens because they were considered to be critical structures. RESULTS: These studies demonstrated that the lower energy photons generated by Pd-103 seeds (average 21 KeV) in ophthalmic plaques were more rapidly absorbed in tissue than photons generated by I-125 (average 28 KeV). Therefore, during ophthalmic plaque radiotherapy, Pd-103 photons were found to be more rapidly absorbed within the tumor and less likely to reach most normal ocular structures. On average, the use of Pd-103 decreased the dose to the fovea by 5.7%, to the optic nerve by 8.4%, to the lens by 26%, and to the opposite eye wall by 38.4%. CONCLUSION: Palladium-103 ophthalmic plaque brachytherapy resulted in slightly more irradiation of the tumor and less radiation to most normal ocular structures. PMID- 8244815 TI - Radiation therapy salvage of Hodgkin's disease following chemotherapy failure. AB - PURPOSE: This study evaluates the efficacy of comprehensive salvage radiation therapy, with or without chemotherapy, in selected patients with Hodgkin's disease who have failed primary combination chemotherapy. METHODS AND MATERIALS: Between 1972 and 1991, 28 patients with advanced Hodgkin's disease failing intensive combination chemotherapy in previously unirradiated nodal and/or pulmonary sites were treated with salvage radiotherapy with curative intent. Treatment consisted of comprehensive extended field radiotherapy to all known areas of disease. Total radiation doses ranged from 17.0 to 50.0 Gy, with only five patients (18%) receiving < 30.0 Gy to any field. Twelve patients also received chemotherapy as part of their salvage regimen. RESULTS: Twenty-six patients (93%) achieved a complete response and 14 (50%) remain in continuous complete remission at 14 to 133 months. In addition, three patients (11%) who failed salvage radiotherapy are now without evidence of disease following additional therapy. With a median follow-up of 47 months (range 14+ to 198+), actuarial median relapse-free survival and overall survival are 46 and 97 months, respectively. Actuarial 5-year relapse-free survival and overall survival are 40% and 63%, respectively. Patients with an initial complete response to chemotherapy had a significantly longer actuarial relapse-free survival than those with an initial partial response (p = 0.02). Salvage therapy was generally well-tolerated and resulted in no treatment-related deaths. CONCLUSION: Comprehensive salvage radiotherapy is of significant benefit in selected patients and should be considered an option for patients with advanced Hodgkin's disease following chemotherapy failure. PMID- 8244816 TI - Radiation treatment of heterotopic bone formation in the temporomandibular joint articulation. AB - PURPOSE: The efficacy and toxicity of radiation therapy used for preventing re formation of heterotopic bone involving the temporomandibular joint are assessed. METHODS AND MATERIALS: Ten patients (15 TMJs) with bony ankylosis of the TMJ were referred after reconstruction with costochondral graft, gap arthroplasty, or debridement of heterotopic bone. Treatment consisting of 10 Gy was delivered early postoperatively to a field encompassing the TMJ with adequate margin. Response to therapy was assessed by comparison of routine roentgenograms obtained preoperatively, immediately postoperatively, and at last follow-up; the Turlington-Durr grading system was used. Median duration of postoperative follow up was 19 months. RESULTS: Radiation therapy prevented ectopic bone re-formation in 10 (69%) of 15 TMJs with prior bony ankylosis. Of the 15 TMJs, 13 (87%) had improvement in their Turlington-Durr scores compared with the preoperative scores. Development of ectopic bone formation was prevented in 9 (90%) of 10 TMJs rendered Turlington-Durr grade 0 postoperatively. Eight of the 10 patients have remained asymptomatic. Treatment was well tolerated. The only complication experienced was parotitis in three patients. CONCLUSION: Radiation therapy is useful for prevention of heterotopic bone redevelopment after TMJ operation. We recommend 10 Gy in 5 fractions beginning early postoperatively for high-risk patients. This strategy appears beneficial in this young patient population, who suffer significant pain and functional impairment in the TMJ articulation. PMID- 8244817 TI - Weekly cisplatin plus external beam radiotherapy and high dose rate brachytherapy in patients with locally advanced carcinoma of the cervix. AB - PURPOSE: Prospective, single arm, Phase I/II trial performed to assess the efficacy and toxicity of the concomitant use of weekly cisplatin and pelvic radiotherapy in patients with locally advanced carcinoma of the cervix. METHODS AND MATERIALS: Between December 1988 and April 1991, 50 previously untreated patients with bulky, locally advanced, squamous cell carcinoma entered the study. All patients were evaluated by a gynecologist and a radiation oncologist and were submitted to standard pre-treatment staging procedures. The International Federation of Gynecology and Obstetrics stage distribution was as follows: IIA three patients, IIB seventeen, IIIA two, IIIB 25, and IVA three. Radiotherapy consisted of 46 Gy external beam irradiation plus three high dose rate intracavitary treatments given on a weekly basis to a total dose of 30 Gy to point A. Cisplatin 30 mg/m2 was also given weekly starting on day 1 of radiotherapy. RESULTS: With a median follow-up time of 27 months, complete response was seen in 88% (44/50) of the patients. The actuarial survival rate at 44 months was 65%. Total pelvic failure rate was 26% (13/50). Of the 44 patients who achieved a complete remission, only seven have failed in the pelvis. Distant disease was observed in 24% of the cases. Treatments were well tolerated with no patient requiring an interruption in the radiotherapy. However, the incidence of late gastrointestinal toxicity was high, with 10 patients developing a rectal ulcer (four colostomies for severe bleeding), two patients a small bowel obstruction, and two patients a recto-vaginal fistula. Moreover, gastrointestinal complications appeared sooner than expected, at a median follow-up time of 11 months after completion of treatment. CONCLUSION: The combination of weekly cisplatin and radiotherapy appears to be a very effective regimen for patients with locally advanced carcinoma of the cervix, but resulted in a relatively high frequency of late gastrointestinal complications. PMID- 8244818 TI - Salvage whole abdomen radiation therapy: its role in ovarian cancer. AB - PURPOSE: In spite of high initial response rates, many patients with epithelial ovarian carcinoma eventually fail their primary treatment. Further treatment with second-line regimens has been ineffective in producing durable responses. Thus, whole abdomen radiation therapy was evaluated as a salvage therapeutic modality as to its feasibility, efficacy, and toxicity. METHODS AND MATERIALS: Between June 1983 and June 1990, 44 patients who failed one or more chemotherapeutic regimens were treated with whole abdomen radiation therapy. Forty patients had epithelial carcinoma of the ovary and the remaining had primary adenocarcinoma of the peritoneal cavity. Radiation was delivered with an open-field technique and 2500 cGy were planned to the whole abdomen, with a boost when indicated. Prior to radiation, the amount of residual disease after debulking was noted to be microscopic in one-half of the patients and macroscopic in the other half. Pelvis alone was the site of residual disease in 14 patients, and upper abdominal involvement was found in 30. RESULTS: Five patients (11%) were unable to complete the planned therapy secondary to acute toxicity. The 4-year actuarial survival and recurrence-free survival rates for the entire group were 23% and 22%, respectively. The survival and recurrence-free survival rates for the group with microscopic residual disease at 37% and 42% were significantly better than those for the patients with macroscopic residual disease at 9% and 5% (p < 0.005; p < 0.001) at 4 years, respectively. Patients with disease limited to pelvis only had a recurrence-free survival of 56% compared to 0% when the upper abdomen was involved (p < 0.005). The abdomino-pelvic cavity was the first site of recurrence in 28 of 31 patients in whom the site of recurrence could be determined. Eight patients (18%) experienced bowel complications, of whom five needed surgical intervention. CONCLUSIONS: Whole abdomen radiation therapy with a pelvic boost is feasible with acceptable acute and late toxicity. It is effective in patients with minimal residual disease. PMID- 8244819 TI - Adjuvant postoperative accelerated hyperfractionated radiotherapy in rectal cancer: a feasibility study. AB - PURPOSE: To assess the acute toxicity and hence feasibility of postoperative hyperfractionated accelerated radiotherapy in rectal cancer. METHODS AND MATERIALS: Twenty patients were submitted to accelerated hyperfractionated radiotherapy after resection of rectal cancer. A total dose of 48 Gy was given in 3 weeks. Two fractions of 1.6 Gy were used with a mean interfraction interval of at least 6 hours. The pelvic volume was treated by a four-field box technique using a linear accelerator (6-18 MV). Acute toxicity was assessed once per week. Small bowel and skin toxicity were scored according to the criteria of the World Health Organization. Bladder toxicity was scored according to the criteria of the Radiation Therapy Oncology Group. RESULTS: All the patients underwent the treatment as planned except one. No patient presented grade 3 or 4 bladder toxicity. There was only one patient who complained from grade 3 skin toxicity at the end of the treatment. Fourteen patients had some degree of intestinal toxicity. This was the most frequently occurring acute side-effect. Only two out of the fourteen patients had intestinal toxicity exceeding grade 2. CONCLUSION: Hyperfractionated accelerated radiotherapy on a pelvic volume is feasible as far as acute toxicity is concerned. PMID- 8244820 TI - The relationship between proliferative and oxygenation status in spontaneous canine tumors. AB - PURPOSE: Immunocytochemical markers have been applied to biopsy specimens from spontaneous canine tumors to assess the prevalence and spatial distribution of proliferating and hypoxic cells, and their "geographic" relationship to each other. Both types of cells have been implicated in the failure to locally control human tumors treated with radiation and chemotherapy. METHODS AND MATERIALS: For the detection of hypoxic cells, a rabbit polyclonal antibody raised against a protein-bound, hexafluorinated, 2-nitroimidazole, designated CCI-103F, was used. The unmetabolized drug must first be injected into the dog to allow time for hypoxic metabolism and cellular binding to occur. For the detection of proliferating cells, a mouse monoclonal antibody raised against an endogenous nuclear protein, the "proliferating cell nuclear antigen," or PCNA, was used. This protein is expressed in most actively proliferating cells, but not in quiescent ones. An indirect immunostaining technique was used to visualize these markers in the tissue sections, and image analysis was used to estimate the area fraction of positive staining in representative, low magnification microscope fields. RESULTS: Tumors with both high and low hypoxic and proliferative area fractions have been identified. No systematic relationship between the prevalence of the two markers, nor of the relationship between tumor grade and proliferative fraction, could be established. Staining with the proliferation marker was more commonly found near blood vessels, but some "nests" of tumor cells apparently distant from vasculature contained many proliferating cells. Staining with the hypoxia marker tended to be distant from the vasculature and/or bordering regions of tumor necrosis, but some labeled cells appeared near blood vessels, and in the absence of necrosis. Staining of sequential sections, one with the proliferation marker and one with the hypoxia marker, indicated that the two cell populations overlapped to varying extents. Some incidental staining of canine normal tissues with both the proliferative and hypoxia markers was observed as well. CONCLUSION: The immunochemical marker approach promises to be a useful tool to increase both our basic understanding of tumor physiology and the complex nature of tumor heterogeneity. PMID- 8244821 TI - Radiation resistance of primary clonogenic blasts from children with acute lymphoblastic leukemia. AB - PURPOSE: Detailed comparative analyses of the radiation sensitivity of primary clonogenic blasts from children with acute lymphoblastic leukemia (ALL) were performed to achieve a better understanding of clinical radiation resistance in ALL. METHODS AND MATERIALS: The radiation sensitivity of primary clonogenic blasts from 74 children with newly diagnosed acute lymphoblastic leukemia (ALL) was analyzed using leukemic progenitor cell (LPC) assays. Primary bone marrow blasts from all 74 patients were exposed to ionizing radiation and subsequently assayed for LPC-derived blast colony formation. Radiation survival curves of primary clonogenic blasts (i.e., LPC) were constructed for each of the newly diagnosed patients using computer programs for the single-hit multitarget as well as the linear quadratic models of cell survival. RESULTS: A marked interpatient variation in intrinsic radiation sensitivity was observed between LPC populations. The SF2 values ranged from 0.01 to 1.00 (median: 0.36; mean +/- SE = 0.40 +/- 0.03), and the alpha values ranged from 0.00 Gy-1 to 3.27 Gy-1 (median: 0.280 Gy-1; mean +/- SE = 0.43 +/- 0.09 Gy-1). Patients were divided into groups according to their sex, age, WBC at diagnosis, cell cycle distribution of leukemic blasts, and immunophenotype. Only immunophenotype provided a significant correlation with the intrinsic radiation sensitivity of LPC. Patients with B lineage ALL had higher SF2 (0.47 +/- 0.04 vs. 0.31 +/- 0.05, p < 0.05) and smaller alpha values (0.43 +/- 0.09 Gy-1 vs. 0.65 +/- 0.10 Gy-1, p < 0.05) than T lineage ALL patients, consistent with greater intrinsic radiation resistance at the level of LPC. Notably, 43% of B-lineage ALL cases, but only 27% of T-lineage ALL cases had LPC with SF2 > or = 0.5. Similarly, 66% of B-lineage ALL cases, but only 37% of T-lineage ALL cases had LPC with alpha values < or = 0.4 Gy-1. Combining the two indicators of radiation resistance, we found that only 34% of the B-lineage ALL patients had none of the two parameters in the respective critical regions (alpha < or = 0.4 Gy-1; SF2 > or = 0.5), while 63% of the T lineage patients had none (p < 0.05). In multivariate analyses, the immunophenotypic B-lineage affiliation was the only significant predictor of radiation resistance at the level of LPC. Whether alone or in combination, none of the other variables examined, including sex, age, WBC, in vitro plating efficiency, S-phase index, and proliferation index were significantly correlated with the radiation sensitivity or resistance of LPC. CONCLUSION: These results offer unprecedented evidence for an association between composite immunophenotype (viz., B-lineage ALL vs T-lineage ALL) and radiation resistance that may form a basis for modifying radiation conditioning regimens. PMID- 8244822 TI - Study of biological effects of varying mixtures of Cf-252 and gamma radiation on the acute radiation syndromes: relevance to clinical radiotherapy of radioresistant cancer. AB - PURPOSE: Data for the 30 day bone marrow syndrome (BM-50) and the 6-10 day gastrointestinal (GI-50) syndrome for a one and two fraction schedule and acute and low dose rate irradiation using pure and mixed Cf-252 and photon radiation are presented. METHODS AND MATERIALS: The radiations of Cf-252 is a mixture of neutrons and gamma rays. We total body irradiated Balb/c mice of both sexes with acute Co-60, low dose rate Cs-137 and Cf-252 using a 1 x and 2 x schedule. For low linear energy transfer radiations of Co-60 or Cs-137 there was expected to be an increase in the dose to produce the gastrointestinal and bone marrow syndromes with minimal change for Cf-252 neutrons. However, the radiations from Cf-252 are approximately 65% neutrons and approximately 35% photons and hence some repair may be expected. We further altered the proportion of photons in the Cf-252 radiation field by mixing Cs-137 with the Cf-252 sources and total body irradiated the mice to determine the effects on the syndromes. We determined the effects of mixing Cf-252 neutrons with different proportions of photons on the radiation syndromes. RESULTS: There was increase in BM-50 and GI-50 doses with fractionated or low dose rate photon irradiations and the dose modifying factors were 1.3-1.4 for the GI syndrome and 1.2 for the bone marrow syndrome. For Cf-252 there was minimal fractionation effect for the GI-50 syndrome, which increased by a 1.1 for x 1 vs. x 2 fractions; for the BM-50 syndrome it rose by a 1.1 factor. For LDR Cs-137 the dose for the GI-50 syndrome rose by a 2.2 fold. For mixed neutron-photon radiation of 0%, 15%, 35%, and 65% eta/gamma mixtures, the dose to produce the BM-50 and GI-50 endpoints dropped sharply from 0 to 35% neutrons and remained flat thereafter. CONCLUSION: For major tissues such as the bone marrow and G-I tract, Cf-252 behaved as high linear energy transfer for mixtures of neutrons and gamma rays of approximately 35% neutrons when the radiation were delivered simultaneously at the low dose rates studied. There was little or no additional contribution to the effectiveness of the mixed radiations if neutrons contributed 35% or more of the dose. PMID- 8244823 TI - Localization of the prostatic apex for radiation therapy using implanted markers. AB - PURPOSE: This report concentrates on the localization of the prostatic apex using implantable markers, and a comparison to localization defined by computed tomography (CT) and retrograde urethrography. METHODS AND MATERIALS: Fifteen patients were entered into a prospective trial and scheduled to undergo (a) pelvic CT, (b) retrograde urethrogram, and (c) transrectal ultrasound with placement of radiodense markers. Three markers were implanted: one was placed at the trapezoid area (prostatic apex), and the others placed lateral to the base of each seminal vesicle. The retrograde urethrogram was performed using standard technique. The superior-inferior distance between the apex identified by the marker placed at the prostatic apex and the other studies was measured. RESULTS: CT and urethrogram overestimated the inferior extent of the prostatic apex when compared to the location as defined by the implanted marker. With CT, the average distance from the marker to the CT-defined apex was 0.6 cm (95% C.I.--0.4-0.8 cm). With urethrogram, the average distance from the marker to the urethrogram defined apex was 1.3 cm (95% C.I.--0.7-1.9 cm). When CT and urethrogram were compared, CT was more accurate in identifying the prostatic apex. CONCLUSION: Under ultrasound guidance, radiodense markers have been implanted into the prostate. This has revealed that the apex is localized superior to the apical margin as defined by retrograde urethrogram and CT, and that CT may be more accurate than retrograde urethrogram. In addition, the placement of multiple markers yields spatial information on prostatic position that can be extracted from megavoltage portal images. PMID- 8244824 TI - Immobilization improves the reproducibility of patient positioning during six field conformal radiation therapy for prostate carcinoma. AB - PURPOSE: To determine the magnitude of patient positioning errors associated with six field conformal therapy for carcinoma of the prostate, and to assess the impact of alpha-cradle immobilization on these errors. METHODS AND MATERIALS: The records of 22 patients, treated at two of the treatment facilities within our department, using computed tomography-planned conformal six field therapy for carcinoma of the prostate, were reviewed. At one facility (UCD), patients were routinely treated with immobilization, while at the other (UCSF) no rigid immobilization was used. Portal films of patients treated at both facilities were subsequently reviewed, and the deviation of each portal from the simulation film was determined (simulation-to-treatment variability). In addition, for each patient, the average deviation of each portal film from the average portal film (treatment-to-treatment variability) was determined. RESULTS: The mean and median simulation-to-treatment variability was 0.4 cm for those patients treated with immobilization, versus 0.6 cm for those treated without immobilization. The 90th percentile of simulation-to-treatment variability was 0.7 cm for those patients treated with immobilization, versus 1.1 cm for those not immobilized. There was a significant reduction in the number of portals observed with errors of > or = 0.50 cm (132/201 vs. 37/87, 66% vs. 43%; p < 0.001), 0.75 cm (184/201 vs. 59/87, 92% vs. 68%; p < 0.001), and 1.0 cm (196/201 vs. 74/87, 98% vs. 85%; p < 0.001) for patients treated with immobilization. There was also a significant reduction in the number of patients with treatment-to-treatment variability > or = 0.5 cm (1/10 vs. 8/12; p = 0.01) for patients treated with immobilization. CONCLUSION: The use of immobilization devices significantly reduces errors in patient positioning, potentially permitting the use of smaller treatment volumes. Immobilization should be a component of conformal radiation therapy programs for prostate carcinoma. PMID- 8244825 TI - Pre-installation empirical testing of room shielding for high dose rate remote afterloaders. AB - PURPOSE: Many facilities are acquiring high dose rate remote afterloading units. It is economical that these units be placed in existing shielded teletherapy rooms. Scatter-radiation barriers marginally protect uncontrolled areas from a high dose rate source especially in a room that houses a non-dynamic Cobalt-60 unit. In addition the exact thickness and material composition of the barriers are unknown and therefore, a calculation technique may give misleading results. Also, it would be impossible to evaluate an entire wall barrier by taking isolated core samples in order to assist in the calculations. A quick and inexpensive measurement of dose equivalent using a rented high activity 192Ir source evaluates the barriers and locates shielding deficiencies. METHODS AND MATERIALS: We performed transmission calculations for primary and scattered radiation based on National Council on Radiation Protection and Measurements Reports 49 and 51, respectively. We then rented a high activity 21.7 Ci (8.03 x 10(11) Bq) Ir-192 source to assess our existing teletherapy room shielding for adequacy and voids. This source was placed at the proposed location for clinical high dose rate treatment and measurements were performed. RESULTS: No deficiencies were found in controlled areas surrounding the room, but large differences were found between the calculated and measured values. Our survey located a region in the uncontrolled area above the room requiring augmented shielding which was not predicted by the calculations. A canopy shield was designed to potentially augment the shielding in the ceiling direction. CONCLUSION: Pre-installation testing by measurement is an invaluable method for locating shielding deficiencies and avoiding unnecessary enhancement of shielding particularly when there is lack of information of the inherent shielding. PMID- 8244826 TI - Advances in 3-dimensional radiation treatment planning systems: room-view display with real time interactivity. AB - PURPOSE: We describe our 3-dimensional (3-D) radiation treatment planning system for external photon and electron beam 3-D treatment planning which provides high performance computational speed and a real-time display which we have named "room view" in which the simulated target volumes, critical structures, skin surfaces, radiation beams and/or dose surfaces can be viewed on the display monitor from any arbitrary viewing position. METHODS AND MATERIALS: We have implemented the 3 D planning system on a graphics superworkstation with parallel processing. Patient's anatomical features are extracted from contiguous computed tomography scan images and are displayed as wireloops or solid surfaces. Radiation beams are displayed as a set of diverging rays plus the polygons formed by the intersection of these rays with planes perpendicular to the beam axis. Controls are provided for each treatment machine motion function. Photon dose calculations are performed using an effective pathlength algorithm modified to accommodate 3-D off center ratios. Electron dose calculations are performed using a 3-D pencil beam model. RESULTS: Dose distribution information can be displayed as 3-D dose surfaces, dose-volume histograms, or as isodoses superimposed on 2-D gray scale images of the patient's anatomy. Tumor-control-probabilities, normal-tissue complication probabilities and a figure-of-merit score function are generated to aid in plan evaluation. A split-screen display provides a beam's-eye-view for beam positioning and design of patient shielding block apertures and a concurrent "room-view" display of the patient and beam icon for viewing multiple beam set ups, beam positioning, and plan evaluation. Both views are simultaneously interactive. CONCLUSION: The development of an interactive 3-D radiation treatment planning system with a real-time room-view display has been accomplished. The concurrent real-time beam's-eye-view and room-view display significantly improves the efficacy of the 3-D planning process. PMID- 8244827 TI - On-line portal imaging: image quality defining parameters for pelvic fields--a clinical evaluation. AB - PURPOSE: A test of several image enhancement techniques, performed on on-line portal images in real clinical circumstances, is presented. In addition a score system enabling us to evaluate image quality on pelvic fields is proposed and validated. METHODS AND MATERIALS: Localization images (n = 546) generated by an on-line portal imaging system during the treatment of 13 patients on pelvic fields were obtained by delivering a radiation dose of 6-8 cGy by an 18 MV photon beam, and recorded with a silicon intensified target video camera with adjustable gain, kV- and black level. Set-up errors were corrected before continuing irradiation. A scoring system based on the number of visible bone-soft tissue edges and transformed to a scale 0 to 5 was developed to judge image quality. A validation of this classification of images was performed with the use of transsectional bone-densities (bone-density*radiological path length) specified at the score defining landmarks. A high pass filter was used on all images, additional on-line open field subtraction was performed on 242 fields. Off-line study was performed in which a panel consisting of two groups (one composed of three radiation oncologists, the other of three radiotherapy technologists), scored 470 pelvic fields without further enhancement, and the same images with Contrast Limited Adaptive Histogram Equalization (CLAHE) (Pizer et al.). Two different clipping levels (3.0 and 5.0) were studied. RESULTS: Gender and transsectional bone-densities were the most defining patient-related factors influencing image quality. Camera settings, gantry angle, and image post processing were important non-patient-related factors. All investigators judged CLAHE to ameliorate low contrast images and to deteriorate good quality images (p < 0.001). PMID- 8244828 TI - Total scalp treatment of mycosis fungoides: the 4 x 4 technique. AB - PURPOSE: A technique for treating mycosis fungoides confined to the scalp using a method known as the 4 x 4 technique is presented. METHODS AND MATERIALS: Uniform dose distribution on the scalp and acceptable "hot spots" along five match lines is accomplished by using four sets of four fields (i.e., 4 x 4) on the patient. Precise and reproducible patient and field alignment was accomplished with a solid thermoplastic mask, which is the surface on which match lines are drawn. In vivo dosimetry (thermoluminescent dosimeters and film) are easily attached to the mask which also provides a portion of the 7 mm bolus used to shift the characteristic 6 MeV electron depth dose toward the skin surface. RESULTS: In vivo dosimetry demonstrated that single fraction match line doses are within 25% to 30% of central axis dose. Shifting these match lines to four locations reduces these "hot spots" to satisfactory levels (less than 120%). Three patients have been treated with this technique and each patient continues to have a complete clinical response at 14 to 21 months post treatment. In addition, each patient has excellent cosmetic results with no evidence of acute or chronic side effects at the match lines. CONCLUSION: The 4 x 4 technique has proven to be useful in the treatment of mycosis fungoides confined to the scalp. PMID- 8244829 TI - Role of bipedal lymphangiogram in radiation treatment planning for cervix cancer. AB - PURPOSE: The role of bipedal lymphangiogram in tailoring radiation portals in radical radiation therapy for Stages II or III cervix cancer is investigated. METHODS AND MATERIALS: The records and simulation films of 87 patients with Stage II or III carcinoma of the cervix treated with radical radiation therapy alone have been retrospectively reviewed. RESULTS: Sixty-two percent of patients who had a bipedal lymphangiogram, subsequently had their radiation fields altered from a "standard portal." The most frequently altered fields were the lateral margin of the postero anterior field and the anterior margin of the lateral fields. In order to cover the lymphatic channels in the pelvis in 90% of cases as outlined by the bipedal lymphangiogram, the lateral margins of the postero anterior fields would need to be 2.5 cm lateral to the pelvic brim and the anterior border of the lateral field, 0.5 cm anterior to the pubic symphysis. CONCLUSION: "Standard" fields adequately cover the median distribution of lymphatics, but may result in a geographic miss in some patients. Bipedal lymphangiograms allow a more accurate tailoring of pelvic lymphatic fields. PMID- 8244830 TI - Inguinofemoral radiation of N0,N1 vulvar cancer may be equivalent to lymphadenectomy if proper radiation technique is used. AB - PURPOSE: To update a previous retrospective study that compared inguinofemoral irradiation (N = 23) to lymphadenectomy (N = 25) for N0,N1 vulvar carcinoma with more patients and longer follow-up. These results, unlike the recent randomized Gynecologic Oncology Group (GOG study), suggest that radiation is a viable alternative to groin dissection, and the differences may be explained on the basis of irradiation technique. METHODS AND MATERIALS: Based on a growing body of data supporting control of subclinical nodal disease with irradiation in vulvar cancer, the decision was made to offer patients with squamous cell carcinoma of the vulva and clinically negative groins (N0,N1), either lymphadenectomy or inguinofemoral irradiation based on clinical factors. Because of the acute skin reaction and possible underdosing of deep femoral nodes with electrons as used in the GOG study, opposed photon fields to 50 Gy were used. From 1983 to 1991, 48 patients underwent a radical vulvectomy followed by either lymphadenectomy (Group I, n = 25) or inguinofemoral irradiation (Group II, n = 23). RESULTS: The actuarial nodal control was 100% in Group I and 91% in Group II (p = 0.14). In addition, there was no difference in cause specific survival at 3 years (96% and 90%, respectively, p = 0.47). The morbidity of lymphadenectomy (Group I) included: 16% lymphedema, 16% seromas, 44% infection, and 68% wound separation. In the irradiated patients (Group II), 16% developed lymphedema and only 9% had a significant skin reaction. CONCLUSION: Based on this analysis of local control and survival with longer follow-up and more patients, irradiation of the N0, N1 inguinofemoral nodes may be a viable alternative to lymphadenectomy for squamous cell carcinoma of the vulva if proper radiation technique and dose are used. In addition, the acute and delayed morbidity of lymphadenectomy exceeds that of irradiation. PMID- 8244831 TI - Femoral vessel depth and the implications for groin node radiation. AB - PURPOSE: To quantify, based on pretreatment computer tomographic measurements, potential groin node depths, which will aid in optimal treatment planning for patients requiring groin node radiation. METHODS AND MATERIALS: The pretreatment computer tomographic scans of 50 gynecologic cancer patients were reviewed to determine the distance of each femoral vessel beneath the overlying skin surface, as an indicator of potential groin node depth. Correlative data regarding height and weight were obtained from patient medical records, and were used to calculate the Quetelet index, defined as (weight in kg)/(height in m)2. Treatment parameters of 5 patients who failed prophylactic groin radiation in a recently published study were assessed to determine if underdosage represented a possible cause of failure. RESULTS: Individual femoral vessel depths ranged from 2.0 to 18.5 cm. When the depths of all four femoral vessels were averaged in each patient, the mean "4-vessel average" depth for this patient population was 6.1 cm. The median Quetelet index for the group was 25.6, and there was a strong correlation between femoral vessel depth and patient Quetelet index. Recalculation of doses provided to the 5 patients failing prophylactic groin radiation in the Gynecologic Oncology Group study showed that all had received potential tumor doses < 4700 cGy, with 3 patients being underdosed by > 30%. CONCLUSION: While surgery is often indicated in the management of patients with potential groin node metastases, the role of prophylactic groin radiation should not be rejected. Data from this study may aid in the optimal design and implementation of groin node radiotherapy. PMID- 8244832 TI - On the lack of demonstrated clinical benefit of neoadjuvant cisplatinum therapy for cervical cancer. AB - PURPOSE: This study ascertains the benefit and detriment of cis-platinum-based regimes for advanced cervical cancer using standard response parameters as well as a methodology to separate potential effects of radiosensitization and effects of control of occult systemic metastases. MATERIALS AND METHODS: In addition to conventional response criteria used to analyze three recently published randomized studies, the efficacy of neoadjuvant chemotherapy is measured with a statistical model to calculate potential improvement in survival offered by perfect local and distant control. RESULTS: Neoadjuvant chemotherapy either did not affect or it impaired survival in the three trials. Evidence of altered failure patterns was obtained. In particular, local control was compromised by neoadjuvant chemotherapy. CONCLUSION: Failure patterns are altered by treatment as well as by standard prognostic factors. Neoadjuvant chemotherapy may decrease survival. Possible reasons include the limited efficacy of cis-platinum alone and in combination with other therapeutic agents, accelerated repopulation caused by neoadjuvant therapy, delay in initiation of radiotherapy by neoadjuvant chemotherapy, low statistical power of published studies, and tumor heterogeneity. Curative modalities such as surgery and radiotherapy must not be compromised by neoadjuvant chemotherapy. PMID- 8244833 TI - Value of 3-D imaging for staging cervical cancers and predicting behavior. PMID- 8244834 TI - The advantage of an X-ray visible marker of the prostatic apex. PMID- 8244835 TI - Groin node irradiation for vulvar cancer: treatment planning must do more than scratch the surface. PMID- 8244836 TI - Radiotherapy in the treatment of vulvar carcinoma. PMID- 8244837 TI - Groin irradiation is a viable option in carcinoma of the vulva. PMID- 8244838 TI - Positive and negative shifts of the readiness potential: preparatory effects. AB - Freude and Ullsperger (1989) reported that positive potentials occurred on some trials in a single trial analysis of the Readiness Potential (RP). The present study investigated the effect of split attention on the occurrence of RPs of different polarities as well as comparing the post-movement potential and auditory-evoked potential after RPs of different polarities. In part 1 of the study, subjects (n = 16) pressed a key in a self-paced manner, concentrating on the movement. In part 2 they were asked to discriminate between two randomly ordered tones, triggered by the key press (with a delay of 560 ms), and to announce the result to the experimenter verbally. The results of part 1 matched previous findings, in that in 41% of all trials RP was positive. In part 2 there was a significant increase in the relative frequency of positive RPs to 54%. This resulted in the magnitude of the averaged RP decreasing from part 1 to part 2. The early component of the averaged RP, which had begun about 1300 ms before the movement in part 1 of the study, disappeared in part 2, where the averaged RP began only 350 ms before the movement. The post-movement potential was significantly larger when the preceding RP was negative. In part 2 of the study we found an N100 in response to the tone; this was larger after positive RPs, but the difference was not significant. The implications of these novel findings for interpretation of the averaged RP are discussed. PMID- 8244839 TI - Event-related potentials and recognition memory: the effects of word imagery value. AB - Event-related potentials (ERPs) were recorded from 11 subjects during a recognition memory task. Subjects were auditorily presented with 160 target words and, after about 10 min, they were engaged in a recognition task. Subjects were required to determine on each of 320 trials if the word was 'new' (never before presented) or 'old' (heard previously). In the recognition test, 160 target words and 160 non-target words were used as stimuli. To assess the effect of word imagery value, 160 stimuli (80 target and 80 non-target) were high imagery words (HI) and the remaining 160 stimuli were low imagery words (LI). Word frequency was balanced across target and non-target words. Results showed that the old/new effect was reflected by a negative component (belonging to the N400 family, Cz maximum) which was larger for new words in the 400-700 ms range. The imagery effect was also present in the 250-550 ms range of the N400, which was larger for HI stimuli. The imagery effect was present at 400-550 ms only for target words. These results confirmed the hypothesis that negative components as N400, could reflect processes occurring during the memory retrieval of items. PMID- 8244840 TI - Comparison of QEEG and response accuracy in good vs poorer performers during a vigilance task. AB - Subjects performed an auditory continuous performance test requiring them to detect targets in a series of letters presented at a rate of 2/s. 2-min samples of EEG were obtained from eight bipolar sites during a resting condition and during early and late (7-10 min) test performance. EEG power spectra from 27 subjects whose performance accuracy decreased between these latter periods (LoVig group) were compared with those from 27 subjects who maintained a constant level of performance (HiVig group). In both groups EEG power changed significantly between resting and test conditions for all frequency bands: beta power increased, especially in fronto-temporal and temporal left-hemisphere sites; alpha and posterior theta decreased; anterior theta and delta increased. Significant changes also were found between early and late test performance: anterior theta and delta power decreased in both groups; temporal beta power decreased in the LoVig group only, and is thus considered the best indicator of performance changes. Other differences found between groups were across conditions. The HiVig group had more anterior beta and less posterior alpha and theta than the LoVig group. EEG results are discussed in relation to an explanation of vigilance errors based on signal detection theory. PMID- 8244841 TI - Difficulty as a determinant of cardiovascular response: moderating effect of instrumentality in an alleviation paradigm. AB - Female subjects performed 40 trials of an easy or difficult digit-recognition task while being exposed to intermittent presentations of a noxious noise. Half (high instrumentality) were told that, if they performed well, there was a high chance that they would work without noise in a subsequent trial period. The rest (low instrumentality) were told that, if they performed well, there was a low chance that they would work without noise in the subsequent trial period. Thus, a good performance insured either a high or low probability of alleviating the aversive stimulus. Results indicated that systolic responsiveness was proportional to task demand when the instrumentality of alleviative behavior was high, but low under both task conditions when the instrumentality of alleviative behavior was low. Reaction time and performance data were generally consistent with the view that these effects were due to group differences in task engagement, or 'active coping'. The main findings conceptually replicate and extend results from two previous studies that crossed difficulty and instrumentality manipulations. They also call further into question familiar theoretical conceptions that intimate direct effects of incentive value and perceived control on cardiovascular reactivity. PMID- 8244842 TI - Autonomic changes in Brahmakumaris Raja yoga meditation. AB - This report presents the changes in various autonomic and respiratory variables during the practice of Brahmakumaris Raja yoga meditation. This practice requires considerable commitment and involves concentrated thinking. 18 males in the age range of 20 to 52 years (mean 34.1 +/- 8.1), with 5-25 years experience in mediation (mean 10.1 +/- 6.2), participated in the study. Each subject was assessed in three test sessions which included a period of meditation, and also in three control (non-mediation) sessions, which included a period of random thinking. Group analysis showed that the heart rate during the meditation period was increased compared to the preceding baseline period, as well as compared to the value during the non-meditation period of control sessions. In contrast to the change in the heart rate, there was no significant change during meditation, for the group as a whole, in palmar GSR, finger plethysmogram amplitude, and respiratory rate. On an individual basis, changes which met the following criteria were noted: (1), changes which were greater during meditation (compared to its preceding baseline) than changes during post meditation or non-meditation periods (also compared to their preceding baseline); (2), Changes which occurred consistently during the three repeat sessions of a subject and (3), changes which exceeded arbitrarily-chosen cut-off points (described at length below). This individual level analysis revealed that changes in autonomic variables suggestive of both activation and relaxation occurred simultaneously in different subdivisions of the autonomic nervous system in a subject. Apart from this, there were differences in patterns of change among the subjects who practised the same meditation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244843 TI - EEG spectral analysis during hypnotic induction, hypnotic dream and age regression. AB - EEG was recorded monopolarly at frontal (F3, F4), central (C3, C4) and posterior (in the middle of O1-P3-T5 and O2-P4-T6 triangles) derivations during the hypnotic induction of the Stanford Hypnotic Clinical Scale (SHCS) and during performance following suggestions of hypnotic dream and age-regression as expressed in the before-mentioned scale. 10 low-hypnotizable and 9 highly hypnotizable and right-handed female students participated in one experimental session. Evaluations were Fast-Fourier spectral analyses during the following conditions: waking-rest in eyes-open and eyes-closed condition; early, middle, and late phases of hypnotic induction; rest-hypnosis in eyes closed condition; hypnotic dream and age regression. After spectral analysis of 0 to 44 Hz, the mean spectral amplitude estimates across seven Hz bands (theta 1, 4-6 Hz, theta 2, 6-8 Hz; alpha 1, 8-10 Hz; alpha 2, 10-13 Hz; beta 1, 13-16 Hz; beta 2, 16-20 Hz; beta 3, 20-36 Hz) and the 40-Hz EEG band (36-44 Hz) for each experimental condition were extracted. In eyes-open and -closed conditions in waking and hypnosis highly-hypnotizable subjects produced a greater 40-Hz EEG amplitude than did low hypnotizable subjects at all frontal, central and posterior locations. In the early and middle hypnotic induction highly-hypnotizables displayed a greater amount of beta 3 than did low hypnotizables and this difference was even more pronounced in the left hemisphere. With posterior scalp recordings, during hypnotic dream and age regression, high hypnotizables displayed, as compared with the rest-hypnosis condition, a decrease in alpha 1 and alpha 2 amplitudes. This effect was absent for low hypnotizables. Beta 1, beta 2 and beta 3 amplitudes increased in the left hemisphere during age regression for high hypnotizables; low hypnotizables, in contrast, displayed hemispheric balance across imaginative tasks. High hypnotizables during the hypnotic dream also displayed in the right hemisphere a greater 40-Hz EEG amplitude as compared with the left hemisphere. This difference was even more evident for posterior recording sites. This hemispheric trend was not evidenced for low hypnotizable subjects. Theta power was never a predictor of hypnotic susceptibility, 40-Hz EEG amplitude displayed a very high main effect (p < 0.004) for hypnotizability in hypnotic conditions by displaying a greater 40-Hz EEG amplitude in high hypnotizables with respect to lows. PMID- 8244844 TI - Bilateral auditory-evoked potentials in conditions of hypnosis and focused attention. AB - Brain event-related potentials (ERPs) evoked by auditory stimulation were used to study cerebral hemispheric activity during hypnosis. ERPs were recorded from bilateral central (C3 and C4) and temporal (T3 and T4) scalp locations in response to tone pips in 6 medium-high and 6 low-susceptible subjects in three conditions: baseline (tones only), hypnosis (tones plus hypnotic induction), and a focused attention control (tones plus a newspaper story read by the hypnotist). Task asymmetries were individually adjusted for baseline asymmetries. Responses from central locations did not differentiate hypnosis from focused attention for either group. The same was true of temporal locations for the low-susceptible group. The predominant temporal lobe pattern for both conditions and groups was larger left than right responses. The exception was the hypnosis condition for the medium-high susceptible group where there was an increase in responses in the right temporal lobe. PMID- 8244845 TI - The law of initial values: a four factor theory. PMID- 8244846 TI - Component analysis of task-related respiratory patterns. AB - The pattern of breathing, determined by time, volumetric and respiratory shape parameters was examined in male students subjected to mental arithmetic tasks, sustained attention tasks, a relaxation task and four levels of graded exercise. Also, the relationships among the respiratory time parameters were determined by performing a factor analysis on a pooled matrix containing the parameter correlations across all experimental conditions. Relative to baseline, the experimental tasks induced rather distinct breathing patterns. During exercise the breathing curves resembled a triangular shape which was caused by a decrease in total breath duration coupled to an increase in both the depth of breathing and the inspiratory duty cycle time. For both the sustained attention and mental arithmetic conditions there was a slight decrease in the depth of breathing and a more pronounced decrease in inspiration time. However, during mental arithmetic the decrease in inspiration was linked to an increase in expiration duration, exhibiting a breathing curve that resembled a 'saw tooth'. In contrast, the attention tasks induced a slight decrease in expiration duration showing a sinusoidal breathing curve which was more similar to the baseline and relaxation breathing pattern. The factor analysis revealed two respiratory factors: a time factor reflecting inspiration, expiration and total cycle duration and a factor related to the inspiratory duty cycle time. We conclude that analysis of breathing patterns with components other than rate and depth of breathing is a potentially useful research tool. PMID- 8244847 TI - Should Mexico hold its horses? PMID- 8244848 TI - Seminar addresses gender issues. PMID- 8244849 TI - "Big difference between being harassed and being offended". PMID- 8244850 TI - Assessing periodontal pockets. PMID- 8244851 TI - Treatment of amitraz toxicosis. PMID- 8244852 TI - New feature prompts question. PMID- 8244853 TI - Autochthonous echinococcosis in a horse. PMID- 8244854 TI - A poison control center for animals: liability and standard of care. PMID- 8244855 TI - Violative drug residues: the record of food animal practitioners looks good. PMID- 8244856 TI - "The human spirit is a marvelous thing.". PMID- 8244857 TI - What is your diagnosis? Bilateral hip dysplasia and possible osteolytic lesion in the left ischium. PMID- 8244858 TI - Demographic and employment shifts of US veterinarians, 1990 and 1992. PMID- 8244859 TI - A brief analysis of the Omnibus Reconciliation Act of 1993. PMID- 8244860 TI - Concentration of alpha 1-acid glycoprotein in dogs with malignant neoplasia. AB - Serum alpha 1-acid glycoprotein (alpha 1AG) concentrations were determined in 55 dogs with previously untreated, histologically confirmed, high-grade lymphoblastic lymphoma, and in 34 dogs with histologically confirmed nonhematopoietic malignancies (13 dogs with carcinomas and 21 dogs with sarcomas). Serum concentrations were again determined in 32 dogs with lymphoma that were in complete remission 3 weeks after 1 dose of doxorubicin (30 mg/m2 of body surface, i.v.) and in 22 dogs that were still in complete remission 3 weeks after a fourth dose of doxorubicin. For comparison, serum alpha 1AG concentrations were measured in 19 clinically normal (control) dogs of similar weight and age. Eight of the control dogs were given 1 dose of doxorubicin (30 mg/m2, i.v.), and serum alpha 1AG concentrations were measured 3 weeks later. In control dogs, mean serum alpha 1AG concentration after treatment with doxorubicin was not significantly different from mean concentration before treatment. Mean alpha 1AG concentrations in untreated dogs with lymphoma, in dogs with sarcomas, and in dogs with carcinomas were all significantly higher than mean concentration for untreated control dogs. In addition, the mean concentration for dogs with osteosarcomas was significantly higher than mean concentration for untreated control dogs. There were no significant differences in mean serum alpha 1AG concentrations among dogs with different clinical stages of lymphoma (stage IIIa, stage IVa, stage Va). However, mean serum alpha 1AG concentrations were significantly increased for dogs with stages IIIa, IVa, and Va lymphoma, compared with mean concentration for untreated control dogs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244861 TI - Radiofrequency ablation of a concealed accessory pathway as treatment for incessant supraventricular tachycardia in a dog. AB - A new technique for the treatment of certain types of cardiac arrhythmias was used in a 3-year-old dog that was evaluated for incessant supraventricular tachycardia (220 to 280 beats/min), which had been refractory to several treatment regimens. The mechanism of supraventricular tachycardia was atrioventricular (AV) reentry, using a dorsoseptal accessory pathway (AP) for retrograde ventriculoatrial conduction (concealed AP). With the dog under general anesthesia and with fluoroscopic monitoring, electrode catheters were introduced into the heart via peripheral vessels. Electrical recordings allowed localization of the accessory AV pathway. Programmed electrical stimulation was used to verify the function of the abnormal AV connection. At the atrial insertion site of the AP, 2 applications of radiofrequency current (45 V, 21.6 W) were delivered to the dorsoseptal right atrium (near the coronary sinus ostium), which eliminated AP conduction and AV reentrant tachycardia. The dog has remained free of tachycardia and has not required medication during more than 1 year of follow-up. PMID- 8244862 TI - Gastritis caused by Aonchotheca putorii in a domestic cat. AB - Aonchotheca putorii is a parasitic nematode of the stomach and small intestines of many wild mammals. Although A putorii has been found in domestic cats in Iowa, it has not been reported to be pathogenic. The parasite caused severe gastric and was associated with a gastric ulcer and secondary anemia in a 10-year-old domestic shorthair cat from Ohio. The source of infection was not determined. Surgical resection of the affected tissue resolved clinical signs. The importance of postoperative administration of an anthelmintic in the resolution of any remaining A putorii is unknown. Ivermectin was given empirically to this cat. It is not known whether, or at what dosage, ivermectin or any other anthelmintic is effective against A putorii. PMID- 8244863 TI - Cutaneous lymphosarcoma and leukemia in a cat. AB - A cat with cutaneous lymphosarcoma and leukemia, similar to Sezary syndrome in human beings, had initial clinical signs that included pruritus and exfoliative dermatosis, associated with weight loss and lymphadenopathy. Dermatopathologic findings and ultrastructural morphologic features of the circulating cells and cellular infiltrate were consistent with Sezary cells. Cutaneous lymphosarcoma and leukemia should be considered in cats with chronic pruritic exfoliative dermatoses. PMID- 8244864 TI - Tethered spinal cord and an intradural lipoma associated with a meningocele in a Manx-type cat. AB - An 8-month-old neutered male Manx-type cat was evaluated because of clear fluid that had been draining from a skin mass dorsocaudal to the sacrum for 1 week. Neurologically, the cat had poor postural reactions and poor withdrawal reflexes in the hind limbs. Ultrasonography of the dorsal sacral area revealed a 3-cm-long hypoechoic tract extending from the skin mass cranioventrally to the area of the sacrum. The tract appeared to contain a circular mass. Results of myelography and computed tomography helped to confirm the diagnosis of a meningocutaneous tract containing a mass. Surgical exploration was performed and the tract was excised. Histologic changes were similar to those in human beings with tethered spinal cord syndrome and an intradural lumbosacral lipoma. Surgery was indicated in this cat to prevent progression of neurologic signs associated with tethered cord syndrome and to prevent problems associated with loss of CSF through a fistulated meningocele. PMID- 8244865 TI - Displacement of the tendon of the superficial digital flexor muscle in dogs: 10 cases (1983-1991). AB - Displacement of the tendon of the superficial digital flexor muscle was diagnosed in 9 dogs (10 tarsi). Four of 10 displacements occurred in Shetland Sheepdogs. All dogs had an acute onset of lameness with swelling over the proximal end of the tuber calcanei. Lateral displacement occurred in 8 tarsi and medial displacement in 2 tarsi. Nonsurgical treatment (exercise restriction, bandaging, and administration of anti-inflammatory medication) was ineffective in the 5 dogs in which it was attempted. Surgical reconstruction of the supporting soft tissues resulted in return to normal function in 9 cases available for follow-up evaluation. PMID- 8244866 TI - Adrenal insufficiency associated with long-term anabolic steroid administration in a horse. AB - Adrenal insufficiency was diagnosed in a 9-year-old American Quarter Horse gelding that had received monthly injections of stanozolol for 8 years. After the injections were abruptly discontinued, the horse developed anorexia, lethargy, weight loss, and bilateral forelimb lameness. Secondary hypoadrenocorticism was diagnosed on the basis of clinical signs, lack of high endogenous plasma ACTH concentration, and lack of cortisol response to administration of ACTH. Because the medical history did not include glucocorticoid administration, the cause was determined to be excessive administration of an anabolic steroid. Treatment consisted of physiologic glucocorticoid replacement for 9 months until adrenal function returned. Findings in this horse indicate that anabolic steroids influence the hypothalamic-pituitary axis in horses. PMID- 8244867 TI - Causes of abortion, stillbirth, and perinatal death in horses: 3,527 cases (1986 1991). AB - Pathology case records of 3,514 aborted fetuses, stillborn foals, or foals that died < 24 hours after birth and of 13 placentas from mares whose foals were weak or unthrifty at birth were reviewed to determine the cause of abortion, death, or illness. Fetoplacental infection caused by bacteria (n = 628), equine herpesvirus (143), fungi (61), or placentitis (351), in which an etiologic agent could not be defined, was the most common diagnosis. Complications of birth, including neonatal asphyxia, dystocia, or trauma, were the second most common cause of mortality and were diagnosed in 19% of the cases (679). Other common diagnoses were placental edema or premature separation of placenta (249), development of twins (221), contracted foal syndrome (188), other congenital anomalies (160), and umbilical cord abnormalities (121). Less common conditions were placental villous atrophy or body pregnancy (81), fetal diarrhea syndrome (34), and neoplasms or miscellaneous conditions (26). A diagnosis was not established in 16% of the cases seen (585). The study revealed that leptospirosis (78) was an important cause of bacterial abortion in mares, and that infection by a nocardioform actinomycete (45) was an important cause of chronic placentitis. PMID- 8244868 TI - Accidental superphosphate fertilizer poisoning in pregnant ewes. AB - Consumption of superphosphate fertilizer by 200 pregnant ewes resulted in signs of toxicosis in 41 ewes, 14 of which died. Predominant clinical signs were marked teeth grinding, voluminous diarrhea, CNS depression, apparent blindness, and a stiff-legged atactic gait. Biochemical abnormalities were hypocalcemia, hypoglycemia, and a high anion gap. The primary toxic principal in superphosphate fertilizers is the fluorine contaminant; however, calcium pyrophosphate and calcium orthophosphate also contribute to toxicosis, which results in acute proximal renal tubular necrosis. Voluntary consumption of superphosphate fertilizer in well-fed livestock is not expected, and was believed to be related to the lack of availability of salt. PMID- 8244869 TI - Small-intestinal volvulus in cattle: 35 cases (1967-1992). AB - Medical records of 35 cattle with small-intestinal volvulus were reviewed. Surgical correction was performed on 32 cattle, and 17 of these cattle were discharged from the hospital. Mean duration of clinical signs for survivors was not significantly different from that for nonsurvivors, and the most commonly recorded clinical signs were abdominal pain, anorexia, lethargy, abdominal distention, and dehydration. Physical examination of cattle with intestinal volvulus revealed tachycardia, tachypnea, and normothermia. Rectal examination findings included distended small intestine, scant feces or mucus, and tight bands coursing dorsoventrally in the middle portion of the abdomen. Clinicopathologic testing revealed azotemia, hypocalcemia, hyperglycemia, and leukocytosis with a left shift. Nonsurvivors had significantly lower mean preoperative venous blood pH and mean base excess and higher mean serum potassium concentration than did survivors. A diagnosis of volvulus of the entire small intestine was made during surgery in 25 cattle, whereas volvulus of the distal jejunum and ileum was diagnosed during surgery in 7 cattle. Survival rate following surgical correction of volvulus of the entire small intestine (44%) was not significantly different from survival rate following surgical correction of volvulus of the distal jejunum and ileum (86%). However, survival rate for dairy cattle (63%) was significantly higher than survival rate for beef cattle (22%). To determine potential risk factors for the development of small-intestinal volvulus, epidemiologic data from cattle admitted to veterinary teaching hospitals throughout North America were collected by searching records entered into the Veterinary Medical Data Base.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8244870 TI - Decision analysis in the evaluation of diagnostic tests. AB - Decision analysis was used to evaluate the clinical usefulness of 2 diagnostic tests: one for canine heartworm disease and the other for bovine traumatic reticulo-peritonitis. Several clinically relevant measures of test performance are introduced, including expected utility, risk profile, testing band, threshold analysis, and the relative cost of misdiagnosis. One of the principal benefits of decision analysis of diagnostic tests is that the technique can be used to determine how changes in underlying assumptions will affect clinical decisions. If clinicians can identify and assign values to relevant variables, then decision analysis can provide clinically meaningful guidelines for interpreting the results of diagnostic tests. To take advantage of these techniques, clinicians must become comfortable with quantitative expressions for test performance, risk, and prognosis. PMID- 8244871 TI - Developments in the national workforce policy: challenges for allied health education. PMID- 8244872 TI - New directions for allied health education accreditation. AB - The American Medical Association's (AMA's) proposal in October 1992 for a new freestanding agency to replace the Committee on Allied Health Education and Accreditation (CAHEA) energized review committees and their sponsoring organizations, educational institutions, and communities of interest to examine the current accreditation system. It also prompted the design of an independent allied health education accrediting body responsive to the needs of allied health professions, educational institutions, students enrolled in allied health educational programs, and the public. The result of a task force and input by many communities of interest, CAHEA's proposed successor agency, the Commission on Accreditation of Allied Health Education Programs (CAAHEP), will be operational in mid-1994 to provide accreditation and related coordination services. It is committed to simplify the existing accrediting process, be more inclusive of allied health professions that provide entry-level education, and serve as a steppingstone for future more far-reaching developments. PMID- 8244873 TI - Faculty research productivity in allied health settings: a TQM approach. AB - Faculty research productivity in colleges of allied health has often been discussed in the literature over the last five years. Articles have focused on the problem of faculty research productivity from various viewpoints, but none have used a theoretical framework to analyze the problem. The total quality management (TQM) framework is currently being used in health care to improve quality and productivity. This article uses the TQM framework to synthesize literature concerning faculty research productivity and verifies the current relevance of synthesis findings using an allied health faculty survey. These analyses show that the TQM framework is useful in suggesting ways to increase faculty research productivity in colleges of allied health. PMID- 8244874 TI - Staffing patterns and practices in programs in communication disorders. AB - Allied health programs in higher education present unique academic and administrative challenges. Clinical education, the practice of providing students with hands-on experience, is frequently the norm rather than the exception. This paper reports results of a study that examined faculty staffing patterns and practices in undergraduate and master's programs in communication disorders. Specifically, the study examined 1) employment of full- and part-time faculty/supervisors, 2) teaching/supervisory responsibilities of faculty/supervisors, 3) degree requirements for clinical supervisors, 4) teaching load for clinical supervisors, and 5) methods for determining supervisory load in clinical teaching. Results address the "traditional" model for determining the number of faculty positions needed and the implications for change in communication disorders. This study is directly related to allocation of faculty resources. PMID- 8244875 TI - Health care reform: Part II. PMID- 8244876 TI - Shaping the future boundaries of occupational therapy. PMID- 8244877 TI - Phenazoviridin, a novel free radical scavenger from Streptomyces sp. Taxonomy, fermentation, isolation, structure elucidation and biological properties. AB - Phenazoviridin is a newly discovered free radical scavenger from microorganisms. It was isolated from the culture of Streptomyces sp. HR04. The structure of phenazoviridin was determined as 6-(3-methyl-2-butenyl)phenazine-1-carboxylic acid 6-deoxy-alpha-L-talopyranose ester on the basis of its spectroscopic and physico-chemical properties. The novel substance showed strong inhibitory activity against lipid peroxidation in rat brain homogenate and exhibited antihypoxic activity in mice. PMID- 8244878 TI - New inhibitors of melanogenesis, OH-3984 K1 and K2. I. Taxonomy, fermentation, isolation and biological characteristics. AB - Melanogenesis inhibitors, OH-3984 K1 and K2 were isolated from fermentation broth of Streptomyces sp. OH-3984. OH-3984 K1 and K2 inhibited the melanogenesis of B16 melanoma cells at concentrations of 7.5 and 3.8 micrograms/ml, respectively, whereas inhibition of tyrosinase activity has not been observed. The microbial metabolites showed no antimicrobiological activities against Gram-positive and Gram-negative bacteria, fungi or yeast at a concentration of 1,000 micrograms/ml. PMID- 8244879 TI - New inhibitors of melanogenesis, OH-3984 K1 and K2. II. Physico-chemical properties and structural elucidation. AB - New melanin synthesis inhibitors, OH-3984 K1 and K2, were isolated from the fermentation broth of Streptomyces sp. OH-3984, and their structures were elucidated by spectroscopic methods and by chemical transformations. OH-3984 K1 (M.W.: 306; C18H26O4) and K2 (M.W.: 308; C18H28O4) have unique gamma-lactone rings, both of which correspond to oxidative products derived from C1-O14 cleavage of the 14-membered lactone group. PMID- 8244880 TI - Novel antitumor antibiotics, saptomycins. I. Taxonomy of the producing organism, fermentation, HPLC analysis and biological activities. AB - Streptomyces sp. HP530 was found to produce novel antitumor antibiotics, saptomycins, closely related to the pluramycin-group and was further found to mutate frequently. The natural mutant produced several new saptomycins as determined by HPLC analyses. We isolated saptomycins A, B, C1, C2 and F from the parent strain and saptomycins D, E, G and H from the mutant. The saptomycins showed antimicrobial activities and potent antitumor activities against human or murine tumor cell lines in vitro and against Meth A fibrosarcoma in vivo. In particular, saptomycin D was most effective component in vivo of all saptomycins. PMID- 8244881 TI - Novel antitumor antibiotics, saptomycins. II. Isolation, physico-chemical properties and structure elucidation. AB - A complex of novel antitumor antibiotics related to the pluramycin-group was isolated from the fermentation of actinomycete, named Streptomyces sp. HP530. The producing strain mutated frequently. The products isolated from the parent strain were designated saptomycins A, B, C1, C2 and F, while those of the mutant were named saptomycins D, E, G and H. These structures were elucidated by several NMR spectral analyses and other spectroscopic experiments. PMID- 8244882 TI - Cyanopeptolins, new depsipeptides from the cyanobacterium Microcystis sp. PCC 7806. AB - Four depsipeptides (peptide lactones), called cyanopeptolins A, B, C and D, have been isolated from the cyanobacterium Microcystis sp. PCC 7806. They possess identical structures consisting of cyclic L-glutamic acid-gamma-aldehyde, L leucine, N-methyl-phenylalanine, L-valine, L-threonine, L-aspartic acid, hexanoic acid and a variable basic amino acid. This variable amino acid can be L-arginine (cyanopeptolin A), L-lysine (cyanopeptolin B), N epsilon-methyl-L-lysine (cyanopeptolin C) and N epsilon,N epsilon-dimethyl-L-lysine (cyanopeptolin D), respectively. The L-glutamic acid-gamma-aldehyde and the amino group of L-leucine form an unusual 3-amino-6-hydroxy-2-oxo-1-piperidine system. L-Threonine is connected to L-valine via its hydroxy-group forming an ester bonding. The hexanoic acid residue is attached to the N-terminal aspartic acid residue which is not a part of the ring structure. The isolation procedure of the four cyanopeptolins as well as structure elucidation are described. Amino acid analysis, GC/MS analysis, FAB-MS and several NMR techniques were used to reveal the structures. PMID- 8244883 TI - Quinolidomicins A1, A2 and B1, novel 60-membered macrolide antibiotics. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological activity. AB - Three novel macrolide antibiotics, quinolidomicins A1, A2 and B1, were isolated from the fermentation broth of Micromonospora sp. JY16. Quinolidomicin A1 inhibited the growth of various tumor cells including multidrug-resistant cells. Quinolidomicin B1 was similarly cytotoxic, while quinolidomicin A2 was inactive against these tumor cells. PMID- 8244884 TI - Quinolidomicins A1, A2 and B1, novel 60-membered macrolide antibiotics. II. Structure elucidation. AB - The structures of novel macrolide antibiotics, quinolidomicins A1, A2 and B1, were elucidated as shown in Fig. 1 by NMR spectral analysis including a variety of two-dimensional techniques. The quinolidomicins possess a novel 60-membered macrolide ring, which is to our knowledge the largest among natural products. PMID- 8244885 TI - Rousselianone A, novel antibiotic related to phenalenone produced by Phaeosphaeria rousseliana. AB - Novel antibiotic, rousselianone A, was produced by fermentation of Phaeosphaeria rousseliana L2144. Rousselianone A was characterized as a new derivative of phenalenone. The antibiotic showed no antimicrobial activity in vitro, but exhibited in vivo antifungal activity against plant diseases with a relatively broad spectrum. PMID- 8244886 TI - A modified method of mixed lymphocyte reaction: establishment of the assay system and its application to extracts of fungal cultures. AB - A modified mixed lymphocyte reaction (MLR) assay was developed to screen immunosuppressive agents. In the MLR, irradiation of splenocytes with UV light was employed for the preparation of stimulator cells, and the highest response was observed with the combination of splenocytes of C3H/He mice as a stimulator and those of C57BL/6 as a responder. The blastogenesis of the responder cells was quantified by a colorimetric method using MTT in 96-well microculture plates. For screening immunosuppressive substances, the MLR in combination with a cytotoxicity test can distinguish immunosuppressors from cytotoxic agents. The applications of this assay system to extracts of fungal culture was also described. PMID- 8244887 TI - Biosynthetic studies of allosamidin. 2. Isolation of didemethylallosamidin, and conversion experiments of 14C-labeled demethylallosamidin, didemethylallosamidin and their related compounds. AB - A new allosamidin analog, termed didemethylallosamidin (3), was isolated from the mycelia of Streptomyces sp. AJ 9463 which is a producer of allosamidin (1) and demethylallosamidin (2). 14C-Labeled 1, 2 and 3 as well as their related compounds, 4, 5, and 6, were prepared to investigate the biosynthesis of 1. Conversion experiments with the labeled allosamidins revealed that 2 was a precursor of 1, but 3 was not incorporated. This suggests that the first N-methyl group is introduced before the cyclization of the aminooxazoline ring during the biosynthesis of 1. Although none of the compounds 4, 5, and 6 were converted to 1, the production of 1 was inhibited by the addition of 4. PMID- 8244888 TI - Microbial modification of pradimicins at C-11 leading to 11-O-demethyl- and 11-O L-xylosylpradimicins A and FA-1. AB - In a screen of pradimicin-nonproducing mutants derived from Actinomadura verrucosospora subsp. neohibisca R103-3, we found a strain capable of producing 11-hydroxyl analogs of pradimicins A and FA-1, designated pradimicins H and FH, respectively. Feeding of pradimicins H and FH to growing cultures of an actinomycete strain AA3798 produced 11-O-L-xylosylpradimicins H and FH, respectively. These 11-O-L-xylosylpradimicins had a broad spectrum of antifungal activity and demonstrated in vivo efficacies against Candida albicans in mice. PMID- 8244889 TI - Stereostructure and NMR characterization of the antibiotic candidin. AB - The stereostructure of the heptaene macrolide antibiotic candidin was established on the basis of NMR studies: 13C, DQF-COSY, ROESY and C,H-COSY experiments. The absolute configuration of the candidin chiral centers were assigned as 3R, 5S, 10R, 11R, 13R, 15S, 16R, 17S, 19S, 34S, 35R, 36R and 37S. PMID- 8244890 TI - Inactivated products of rifampicin by pathogenic Nocardia spp.: structures of glycosylated and phosphorylated metabolites of rifampicin and 3-formylrifamycin SV. AB - Rifampicin (1) was converted into four inactivated products by pathogenic Nocardia, RIP-1 and RIP-2 by N. brasiliensis and RIP-3 and RIP-4 by N. otitidiscaviarum. MS and NMR analysis showed the compounds to be 3-formyl-23-[O (beta-D-glucopyranosyl)]rifamycin SV (2), 23-[O-(beta-D glucopyranosyl)]rifampicin (3), 21-(O-phosphoryl)rifampicin (4) and 3-formyl-21 (O-phosphoryl)-rifamycin SV (5), respectively. PMID- 8244891 TI - A novel compound related to chartreusin from a mutant of Streptomyces chartreusis. PMID- 8244892 TI - RK-397, a new oxo pentaene antibiotic. PMID- 8244893 TI - Structure of RK-397, a new oxo pentaene antibiotic. PMID- 8244894 TI - Mer-N5075A, a potential HIV-1 protease inhibitor, produced by Streptomyces chromofuscus. PMID- 8244895 TI - Metabolic products of microorganisms. 268. Obscurolides, a novel class of phosphodiesterase inhibitors from streptomyces. II. Minor components belonging to the obscurolide B to D series. PMID- 8244896 TI - The effect of 1 beta-methyl and imidoyl substituents on the antipseudomonal activity of carbapenems. PMID- 8244897 TI - Rapid and alternative methods for the detection of salmonellas in foods. PMID- 8244898 TI - Survival of pathogenic bacteria during mesophilic anaerobic digestion of animal waste. AB - The survival of pathogenic bacteria was investigated during the operation of a full-scale anaerobic digester which was fed daily and operated at 28 degrees C. The digester had a mean hydraulic retention time of 24 d. The viable numbers of Escherichia coli, Salmonella typhimurium, Yersinia enterocolitica, Listeria monocytogenes and Campylobacter jejuni were reduced during mesophilic anaerobic digestion. Escherichia coli had the smallest mean viable numbers at each stage of the digestion process. Its mean T90 value was 76.9 d. Yersinia enterocolitica was the least resistant to the anaerobic digester environment; its mean T90 value was 18.2 d. Campylobacter jejuni was the most resistant bacterium; its mean T90 value was 438.6 d. Regression analysis showed that there were no direct relationships between the slurry input and performance of the digester and the decline of pathogen numbers during the 140 d experimental period. PMID- 8244899 TI - Antimicrobial effect of chlorine on Yersinia enterocolitica. AB - The effects of chlorine at varying pH, culture media and incubation temperatures on one type and two wild type strains of Yersinia enterocolitica were studied. Exposure to 1 and 5 mg 1(-1) did not diminish viability, even after prolonged exposure. A level of 10 mg 1(-1) was required to achieve a 5-log reduction in 120 s for the type strain and 80 s for the wild strains. There was an increase of more than 30% in the rate of disinfection with a 10 degrees C rise, a remarkable increase in antimicrobial activity at pH 5-log reduction in 20 s, as well as marked neutralization of the effect in the presence of 0.1% peptone. Younger cells were more susceptible than older ones, and those from liquid medium more resistant than those from solid medium. Incubation temperature of a 24-h inoculum failed to show any influence. Lastly, there was a noteworthy demand for free chlorine by bacterial biomass, with agreement of the curve depicting the drop in free chlorine in the presence of inoculum with biphasic kinetics of survival curves. PMID- 8244900 TI - Microflora of the aerobic preservation of directly brined green olives from Hojiblanca cultivar. AB - New procedures for the preservation stage of ripe olives from Hojiblanca cultivar were studied. An aerobic fermentative process was used with initial pH correction (0.3% acetic acid) and various NaCl concentrations: 6, 3 and 0% (w/v) in tap water. Treatments were carried out at industrial level and the spontaneous changes monitored. At initial salt concentrations of 6 and 3% (w/v) NaCl, pH rose progressively, reaching 4.3 at equilibrium maintaining during this period a constant free lactic acidity of around 0.4% (w/v). When the initial solution was tap waste, however, the pH decreased rapidly to stabilize at about 3.7, and lactic acidity increased continuously to reach values over 1% (w/v) at the end of the preservation process. In all treatments aeration effectively purged the carbon dioxide from the preservation brines, preventing shrivelling of olives. The microbial growth was strongly influenced by the initial NaCl concentration. At 6 and 3%, only yeasts grew, the most abundant being Pichia membranaefaciens, P. vini, P. fermentans and Hansenula polymorpha. However, when there was no NaCl, lactic acid bacteria colonized the solution. Lactobacillus plantarum and Pediococcus inopinatus were the only species found. In this case there was a co existence between yeasts and lactic acid bacteria. As the treatment that supported lactic acid bacteria achieved the best final pH and acidity for olive stability, it may help to overcome the obstacles to a lactic fermentative process during the preservation stage of ripe olives from the Hojiblanca cultivar. PMID- 8244901 TI - Polymerase chain reaction for the rapid identification of Clostridium botulinum type A strains and detection in food samples. AB - A polymerase chain reaction (PCR) was developed for the detection of Clostridium botulinum type A, a cause of human botulism. A two primer set and an oligonucleotide detection probe were used to specifically detect Cl. botulinum type A neurotoxin gene (BoNT/A). After 40 cycles of amplification, detection of a 798 bp amplified DNA fragment was carried out by agarose gel electrophoresis and Southern blot hybridization. This assay was able to detect 12.5 fg of purified target DNA or five bacteria per reaction. The sensitivity in artificially contaminated food samples after an 18 h enrichment step ranges from 10 to 10(3) bacteria per g according to the type of food samples. No cross-reactions were observed with the other Cl. botulinum toxinotypes and other bacteria found routinely in food. This PCR method may provide a suitable and rapid alternative to standard techniques for detection of Cl. botulinum type A in food samples. PMID- 8244902 TI - Study of Enterobacteriaceae throughout the manufacturing and ripening of hard goats' cheese. AB - The evolution of the counts and the species of Enterobacteriaceae as well as some physico-chemical parameters (pH, aw and NaCl and moisture contents) during manufacturing and ripening of a hard Spanish goats' cheese of the Armada-Sobado variety were studied. Enterobacteriaceae (mean log counts 4.45 g-1 in milk) increased 0.71-2.18 log units in curd and afterwards decreased until they disappeared after 2-4 weeks of ripening. This premature disappearance seems to be due to the decrease in aw values and in moisture contents. However, the low pH values, reached from the beginning of the ripening process, could also contribute to this phenomenon. The most abundant species in milk was Serratia liquefaciens (57.5% of isolates), followed by Morganella morganii (27.5%), Hafnia alvei (5%), Klebsiella oxytoca (5%) and Yersinia enterocolitica (5%). Yersinia enterocolitica was not subsequently isolated from either curd or in cheese. Hafnia alvei numbers increased in curd and in 1-week-old cheese where this micro-organism was the most abundant (47.5% and 75% of the isolates respectively). Escherichia coli, which was not isolated from milk, curd or 1-week-old cheese, was the predominant organism in 2-week-old cheese (57.8% of isolates). This confirms the finding of other authors who have shown that it is one of the most resistant species in ripening cheeses. PMID- 8244903 TI - Fluorescent detection-polymerase chain reaction (FD-PCR) assay on microwell plates as a screening test for salmonellas in foods. AB - This study evaluates a polymerase chain reaction assay coupled with a fluorescent detection in microwell plates for salmonellas in food samples. Chelex 100 extracted cultures and bulk and processed food samples were used as templates for a PCR assay in microwell plates, with a primer pair that amplifies a 206 bp segment of IS200. The PCR products were then denatured by heat and transferred to CovaLink NH plates (Nunc) to which capture oligonucleotides were covalently bound. Hybridization was performed for 1 h at 55 degrees C, the microwells were washed and an alkaline phosphatase-labelled probe, complementary of an internal sequence of the PCR product, was added. After stringent washes, 100 microliters of 1 mmol 1(-1) AttoPhos (JBL Scientific) was then added to the wells and the fluorescence measurement system (Millipore). The level of detection of the assay was as low as 1-10 cfu. A total of 172 food samples were tested, both by culture and FD-PCR. Of these 53 were culture positive and 119 culture negative. The sensitivity of the FD-PCR assay was 100% and the specificity was 90.1%. Positive and negative predictive values were 82.8 and 100%, respectively. Based on the results obtained in this study it appears that the FD-PCR assay described here can be useful to screen a large number of food samples for contamination by salmonellas. PMID- 8244904 TI - Numerical classification and identification of Acinetobacter genomic species. AB - A total of 211 Acinetobacter strains (representing all currently recognized genomic species) were tested for 329 biochemical characters. Overall similarities of all strains were determined for 145 characters by numerical taxonomic techniques, the UPGMA algorithm and the S(SM)) and the S(J) coefficients as measures of similarity. Seven clusters (two or more strains) and three unclustered strains were recovered at a similarity level of 80.0% (S(SM). At this level a complete correspondence between phenotypic cluster and genomic species was found only for genomic species 12 (Ac. radioresistens). At higher similarity levels (84.0% to 84.6% (S(SM)), however, several subclusters were found, each representing a single genomic species. An exception were the strains belonging to the genetically closely related species of the Acinetobacter calcoaceticus baumannii complex. These were recovered scattered in several subclusters. The degree of genomic relatedness between some DNA groups correlated with phenotypic similarities, especially for DNA group 8 (Ac. Iwoffii) and 15 of Tjernberg and Ursing, and for DNA group 4 (Ac. haemolyticus) and 6. For the majority of genomic species, two identification matrices were constructed consisting of 22 and 10 diagnostic characters, respectively. The correct identification rates for the matrices were 98.0% (22 tests) and 90.8% (10 tests) taking a Willcox probability > 0.9. For unambiguous identification of some genomic species, however, additional methods (preferably DNA-DNA hybridization or ribotyping) should be used. PMID- 8244905 TI - Lactobacillus cells in the rabbit digestive tract and the factors affecting their distribution. AB - Lactobacillus fermentum was present in small numbers in the caecum and colon of only one of 16 rabbits. Studies aimed at elucidating the factors affecting the colonization of lactobacilli in rabbit gut were thus performed. These studies included investigation of the effect of gastric juice and bile salts on the viability of lactobacilli, assay of the survival rates of lactobacilli in the gastrointestinal tracts of the rabbits with or without ileum cannulation and measurement of the adhesive capability of lactobacilli to the rabbit intestinal epithelial cells. Results showed that, although some lactobacilli were resistant to the rather low pH levels of rabbit gastric juice, lack of adhesive capability may prevent them from colonizing in the intestinal tract. Therefore, if lactobacilli are to be used as probiotics for rabbit, factors that affect their viability and ability to colonize should be considered. PMID- 8244906 TI - Prognostic factors in childhood acute lymphoblastic leukemia. AB - Treatment efficacy alters the impact of most prognostic factors. Among clinical features, only age and leukocyte count remain prognostically important. Immunophenotyping is useful for ALL classification and for assignment to specific therapy regimens, but, with the possible exception of CD10 expression, has little prognostic importance in the context of contemporary phenotype- and risk-directed therapy. Cytogenetic features are useful for risk assignment. Hyperdiploidy > 50 chromosomes is associated with a favorable prognosis, whereas Ph+ chromosome and t(4;11) confer an adverse prognosis. Pre-B cases with the t(1;19) do not fare well with antimetabolite-based therapy and should be treated with additional classes of chemotherapeutic agents. Finally, certain specific rearrangements such as dic(9;12) may in fact be associated with favorable prognosis. With the exception of treatment for B-cell ALL (and perhaps transitional pre-B ALL), phenotype- or genotype-directed therapies have not been successfully devised. Selection of treatment for individual patients, therefore, should be based on their estimated risk of failure. For patients with very high-risk leukemia (i.e., those with > 70% likelihood of treatment failure), the use of experimental therapeutic strategies, despite the potential for acute and long-term disabilities, may be justified. For the subset of children in the lower-risk category (< 20% probability of failure), antimetabolite-based therapy should be employed to minimize long-term sequelae. PMID- 8244907 TI - Monoclonality in reactive lymphadenopathy: gene rearrangement and multiparameter analysis. AB - Multiparameter analysis of lymph nodes with follicular, interfollicular, and/or atypical hyperplasia was undertaken to search for monoclonality. Twenty-three patients aged 7 to 75 years (mean 32 years) were studied. One patient had a history of lymphoma; two were HIV-positive. Nodes were removed for clinical suspicion of lymphoma. Light microscopy revealed increased and&or abnormal follicular proliferation and occasional progressive transformation of germinal centers. Immunostaining of frozen sections revealed CD4, CD8, kappa, and lambda positivity with more CD4+ than CD8+ cells. Flow cytometry showed a mixed population of T and B cells with no evidence of clonality. Hybridization studies with JH and JK probes showed rearranged bands in one case. No rearrangements were seen with CT beta and bcl-2 probes. Follow-up of 3 to 5 years showed no new occurrences of lymphoma. Although no evidence of monoclonality was seen with other parameters, DNA hybridization revealed heavy and light chain gene rearrangement in 4% (1 of 23). PMID- 8244908 TI - Combination therapy with rhGM-CSF and rhEpo for two patients with refractory anemia and aplastic anemia. AB - Because GM-CSF possesses burst-promoting activity (BPA) and megakaryocyte colony stimulating activity (Meg-CSF) as well as stimulating activity on granulocyte macrophage progenitors, and erythropoietin (Epo) has thrombopoietin-like activity, the combination therapy of GM-CSF and Epo seems to be more effective for stimulating erythropoiesis and thrombocytopoiesis in patients with pancytopenia. For this reason, the combination therapy of recombinant human GM CSF (rhGM-CSF) and rhEpo was performed in two patients with refractory anemia (RA) and aplastic anemia (AA). Epo-unresponsive anemia was remarkably improved by adding rhGM-CSF to Epo and the effect lasted for 1 1/2 months in a patient with RA, but severe anemia occurred again immediately after the discontinuation of Epo. The neutralizing antibodies against GM-CSF were not demonstrated at the phase when anemia re-progressed in this patient. In a patient with AA, anemia and thrombocytopenia, which were refractory to previous administration of rhGM-CSF, responded to the combined administration of GM-CSF and Epo. Although the effects were maintained for 3 1/2 months, the anemia and thrombocytopenia became worse again after the administration of rhGM-CSF was changed from daily to every other day. These findings suggest the usefulness of combination therapy of GM-CSF and Epo for patients with pancytopenia. PMID- 8244909 TI - Molecular biology of chromosomal aberrations in leukemia/lymphoma. PMID- 8244910 TI - In memoriam Jose M. Trujillo, M.D. 1925-1992. PMID- 8244911 TI - Asthma in the inner city--a growing public health concern in the United States. PMID- 8244912 TI - Characteristics of asthma mortality and morbidity in African-Americans. AB - The percent rise in the number of asthma deaths was analyzed using data from the National Center of Health Statistics and compared for African-Americans and Caucasians. The rate of increase for African-Americans in the period 1979-1983 was nearly twice that of Caucasians, and the difference among genders for Caucasians was significantly higher for females. In Baltimore a high percentage (29%) of adult asthma patients (86.8% African-American) seen in an emergency room (ER) and living in the inner city had frequent visits (6 or more annually) to the ER. One-third of the patients used the ER exclusively for asthma management, and 39% delayed for at least 48 hr after onset of symptoms before seeking medical assistance. One-fourth had daily symptoms, and 11% of those regularly employed had missed 10 or more days annually because of asthma. Among the high ER users, 39% required more than one annual hospitalization for management of acute exacerbation of asthma symptoms. Risk factors for mortality and morbidity among inner-city and minority populations as well as potential areas of intervention are discussed. PMID- 8244913 TI - Ease-of-use study of pirbuterol acetate in the Autohaler actuator in three countries: the United States, Germany, and France. AB - The preferred form of acute bronchodilator therapy is to administer aerosolized beta-agonist by a metered-dose inhaler. Many patients cannot use this device efficiently, the major problem being coordination of dose release with inspiration. A new breath-actuated inhaler (the Autohaler actuator) has been developed to overcome this problem. The objective of this study was to evaluate patient's and staff's perceptions of this device. Ninety-eight percent of patients rated the device easy to use and 83% rated the overall use as excellent or good. The staffs rated it as excellent or good for 93% of patients. It was concluded that patients would accept this device, and greater patient compliance could be realized. PMID- 8244914 TI - Sensitization of asthmatic children to common environmental allergens according to their residence. AB - The medical records of 974 asthmatic children aged 1-14 years (mean 8.7 +/- 3.9 years) who had been evaluated with skin prick tests (SPT) in two referral Children's Hospitals in Athens from 1975 to 1987 were analyzed. The children were grouped according to their residence into groups from urban area (UR), rural area (RU), and coastals (CO). The prevalence of positive SPT and the sensitizing allergens according to the residential area and the family atopic history were considered. It was found that 662/974 (68%) children had positive SPT with 63.6%, 70.7%, and 80.4% in UR, RU, and CO respectively. There was a significant difference in the prevalence of positive SPT between UR and CO. A positive family atopic history was more often accompanied by positive SPT in UR only. Sensitization to grass pollens was noted with higher prevalence in UR. The house dust mite Dermatophagoides pteronyssinus sensitization was more prevalent in CO. Our results support the notion that the environment can influence the prevalence of sensitization to common environmental antigens, the kind of sensitizing allergen, and the expressiveness of the genetic factor with regard to development of atopic asthma. PMID- 8244915 TI - Cryopreservation of human bronchi. AB - Human bronchi have been investigated in vitro without or after storage at -196 degrees C in different media containing 1.8 M dimethyl sulfoxide and 0.1 M sucrose as cryoprotectants, dissolved in either fetal calf serum (FCS), Krebs Henseleit solution (KH), or 50% FCS in KH as vehicles. As assessed by the post thaw responses to both carbachol and histamine, optimal preservation of contractile responsiveness was obtained with bronchi that had been frozen in a medium containing KH solution as the vehicle. With each group of cryopreserved bronchi, maximal responses to relaxant agonists such as isoprenaline, papaverine, and the potassium channel activator bimakalim were attenuated by up to 50%, and the passive resting tension after maximal pharmacological relaxation was considerably higher than in the unfrozen tissues. The evidence suggests that, despite some reduction in elasticity, cryopreservation of human bronchi at -196 degrees C preserves both contractile and relaxant mechanisms and offers clear potential for storing human airway smooth muscle for subsequent pharmacological experiments. PMID- 8244916 TI - Objective and subjective bronchodilation over 12 hours after inhaled formoterol: individual responses. AB - The aim of the present study was to compare the duration of bronchodilation obtained with 12 micrograms formoterol and 24 micrograms formoterol with that of 200 micrograms salbutamol over 12 hr. Thirteen stable asthmatics (mean age 59 years, asthma duration 7 years, mean FEV1 56% predicted, and reversibility to beta-adrenoceptor stimulation 36%) were studied in a double-blind crossover study. Lung function test (FEV1 and FVC), heart rate, tremor, and subjective effects were recorded before and 30 min after the test doses and every hour up to 12 hr. The test doses were randomized and given double-blindly as two puffs using a spacer. The median duration of effect, defined as time when FEV1 fell below 20% the maximum bronchodilating capacity, was longer than 12 hr for both formoterol doses, whereas it was 7 hr for salbutamol (p < 0.01). No difference between the two doses of formoterol was seen. The median of the patients' subjective evaluation of the duration of effect was 12 hr, more than 12 hr for 12 and 24 micrograms formoterol, respectively, and 8 hr for salbutamol (p < 0.01). Only 1 patient needed rescue medication on the 2 formoterol days. However, 6 patients inhaled rescue medication during treatment with salbutamol. There were no differences with regard to heart rate, blood pressure, subjective tremor, or palpitations. Formoterol, 12 micrograms and 24 micrograms, was shown to produce at least 12 hr of bronchodilating effect in most patients. However, there was considerable individual variation in duration of effect. PMID- 8244917 TI - A life activities questionnaire for childhood asthma. AB - The paper describes the development and testing of a paper-and-pencil instrument, the Life Activities Questionnaire for Childhood Asthma, that can be applied to assess activity restriction in children with asthma. It describes how items for the instrument were selected to ensure the validity of the developed instrument. The method by which the reliability of the questionnaire was established is also discussed. The result is a valid and reliable instrument that should have wide applicability in measuring activity restriction because of childhood asthma. The developed instrument has potential use in clinical settings, research, and policy making. PMID- 8244918 TI - Effects of glucocorticoids on humoral and cellular immunity and on airway inflammation in patients with steroid-dependent intractable asthma. AB - The effects of glucocorticoids on the proportion of lymphocytes in bronchoalveolar lavage (BAL) fluid in relation to humoral and cellular immunity were studied in 56 patients with steroid-dependent intractable asthma. To analyze the mechanism responsible for reduced numbers of BAL lymphocytes, we divided the subjects into 4 groups according to their BAL lymphocyte proportions: 0-4.9%, 5.0 9.9%, 10.0-14.9%, and 15.0-20.0%. Serum IgG levels and the peripheral lymphocyte count were significantly reduced in patients with a low proportion of BAL lymphocytes (less than 9.9%) than in those with more than 10% BAL lymphocytes. Delayed cutaneous reactivity to purified protein derivative was suppressed in patients with a low proportion of BAL lymphocytes (less than 4.9%). The mean proportion of BAL neutrophils tended to increase as the proportion of BAL lymphocytes decreased. These results show that the reduction in BAL lymphocytes produced by glucocorticoids is associated with suppressed humoral and cellular immunity, and that under such conditions the proportion of BAL neutrophils increases. PMID- 8244919 TI - Transcription-repair coupling and mutation frequency decline. PMID- 8244920 TI - Formation of the K30 (group I) capsule in Escherichia coli O9:K30 does not require attachment to lipopolysaccharide lipid A-core. AB - Escherichia coli K antigens (capsular polysaccharides) are divided into two broad classes, designated groups I and II, on the basis of a number of chemical, physical, and genetic criteria. Group I K antigens can be further subdivided on the basis of the absence (group IA) or presence (group IB) of amino sugars in the repeating unit of the K antigen. One criterion proposed for inclusion in group I is covalent linkage of the capsular polysaccharide to the lipid A-core of lipopolysaccharide (LPS). E. coli O9:K30 is a strain with a representative group IA K antigen. This organism synthesizes an LPS-associated low-molecular-weight form of K30 antigen which is called K(LPS). To determine the involvement of LPS lipid A-core in expression of the K30 capsular polysaccharide, E. coli K30/K-12 hybrid strains were constructed with mutations in the E. coli K-12 rfa locus, responsible for the biosynthesis of the LPS core oligosaccharide. These strains lack K(LPS), indicating that a full-length core is required for K(LPS) expression. However, formation of a K30 capsule was unaffected by rfa defects, indicating that attachment to lipid A-core is not an obligatory step for either export of high-molecular-weight capsular polysaccharide or maintenance of the capsular structure on the cell surface. Silver-stained tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of lipopolysaccharides from other E. coli K serotypes showed that all strains with group IB K antigens expressed some K(LPS). In contrast, some strains with group IA K antigens appear to lack K(LPS). Consequently, although association of group 1 K antigens with lipid A-core is common, it is not a universal marker for inclusion in group I. PMID- 8244921 TI - Genetic and biochemical characterization of the oligopeptide transport system of Lactococcus lactis. AB - The nucleotide sequence of a chromosomal DNA fragment of Lactococcus lactis subsp. lactis SSL135, previously implicated in peptide utilization, has been determined. The genes oppDFBCA, encoding the oligopeptide transport system (Opp), and that encoding the endopeptidase PepO were located on this 8.9-kb DNA fragment. The oppDFBCA and pepO genes are probably organized in an operon. Analysis of the deduced amino acid sequences of the genes indicated that the oligopeptide transport system consists of two ATP-binding proteins OppD and OppF, two integral membrane proteins OppB and OppC, and a substrate-binding protein OppA. On the basis of the homology of OppF and OppD of L. lactis with other ABC (ATP-binding cassette) transporter proteins, the L. lactis Opp system can be classified as a member of this group. Two integration mutants, one defective in OppA and the other defective in PepO, were constructed. Growth of these mutants in a chemically defined medium with oligopeptides showed that the transport system, but not the endopeptidase, is essential for the utilization of peptides longer than three residues. Uptake of the pentapeptide Leu-enkephalin in glycolyzing lactococcal cells was followed by rapid hydrolysis of the peptide intracellularly. Importantly, extracellular hydrolysis of Leu-enkephalin is not observed. The OppA-deficient mutant was unable to transport Leu-enkephalin. Growth experiments with pasteurized milk revealed that transport of oligopeptides forms an essential part of the proteolytic system in lactococci. PMID- 8244922 TI - Use of a reporter gene to follow high-pressure signal transduction in the deep sea bacterium Photobacterium sp. strain SS9. AB - Photobacterium sp. strain SS9 is a deep-sea bacterium which modulates the abundances of several outer membrane proteins as a function of hydrostatic pressure. These proteins include the product of the previously cloned ompH gene (D. H. Bartlett, M. Wright, A. A. Yayanos, and M. Silverman. Nature (London) 342:572-574, 1989). Subsequent to conjugal plasmid delivery it was possible to cross an ompH::lacZ transcriptional fusion into the genome of SS9, replacing the wild-type ompH gene, generating strain EC10. EC10 is not impaired in growth at high pressure, indicating that under the growth conditions employed, OmpH is not required for baroadaptation. beta-Galactosidase production in EC10 is induced by high pressure to approximately the same extent that OmpH production is in the parental strain, SS9. Therefore, OmpH abundance appears to be primarily regulated at the transcriptional level. EC10 was used for the isolation of ompH regulatory mutants. Derivatives of EC10 which produce reduced levels of beta-galactosidase at both low and high pressure and which appeared to possess mutations outside the ompH::lacZ locus were obtained. All of these regulatory mutants displayed alterations in the high-pressure repression of a second outer membrane protein, designated OmpL, and two of the mutants were also deficient in the high-pressure induction of a third outer membrane protein, designated OmpI. The most dramatic phenotype was present in mutant EC1002, whose growth was extremely barosensitive. EC1002 is the first pressure-sensitive mutant ever isolated. Prolonged incubation of EC1002 at high pressure led to the accumulation of cells with wild-type growth characteristics at high pressure. These cells are suggested to possess suppressor mutations, as they remain deficient in beta-galactosidase production and maintain their high-pressure-adapted phenotype for many generations in the absence of high pressure selection. PMID- 8244923 TI - A novel antivirulence element in the temperate bacteriophage HK022. AB - Lysogens of the temperate lambdoid phage HK022 are immune to superinfection by HK022. Superinfection immunity is conferred in part by the action of the HK022 CI repressor at the O.R operators. In this work, we have identified an additional regulatory element involved in immunity. This site, termed OFR (operator far right), is located just downstream of the cro gene, more than 250 nucleotides distant from OR. The behavior of phage containing a mutation in OFR suggests that the wild-type site functions as an antivirulence element. HK022 OFR- mutants were able to form turbid plaques indistinguishable from those of the wild type. However, they gave rise to virulent derivatives at a far higher frequency than the wild type (approximately 10(-5) for OFR- versus about 10(-9) for the wild type). This frequency was so high that cultures of HK022 OFR- lysogens were rapidly overgrown by virulent derivatives. Whereas virulent mutants arising from a wild-type OFR+ background contained mutations in both OR1 and OR2, virulent derivatives of the OFR- mutant phage contained a single mutation in either OR1 or OR2. We conclude that the wild-type OFR site functions to prevent single mutations in OR from conferring virulence. The mechanism by which OFR acts is not yet clear. Both CI and Cro bound to OFR and repressed a very weak rightward promoter (PFR). It is unlikely that repression of PFR by CI or Cro binding to OFR can account in full for the antivirulence phenotype conferred by this element, since PFR is such a weak promoter. Other models for the possible action of OFR are discussed. PMID- 8244924 TI - Characterization of the cell wall of the sheathed methanogen Methanospirillum hungatei GP1 as an S layer. AB - The cell wall of Methanospirillum hungatei GP1 is a labile structure that has been difficult to isolate and characterize because the cells which it encases are contained within a sheath. Cell-sized fragments, 560 nm wide by several micrometers long, of cell wall were extracted by a novel method involving the gradual drying of the filaments in 2% (wt/vol) sodium dodecyl sulfate and 10% (wt/vol) sucrose in 50 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffer containing 10 mM EDTA. The surface was a hexagonal array (a = b = 15.1 nm) possessing a helical superstructure with a ca. 2.5 degrees pitch angle. In shadowed relief, the smooth outer face was punctuated with deep pits, whereas the inner face was relatively featureless. Computer-based two-dimensional reconstructed views of the negatively stained layer demonstrated 4.0- and 2.0-nm wide electron-dense regions on opposite sides of the layer likely corresponding to the openings of funnel-shaped channels. The face featuring the larger openings best corresponds to the outer face of the layer. The smaller opening was encircled by a stalk-like mass from which 2.2-nm-wide protrusions were resolved. The cell wall in situ was degraded at pH 9.6 at 56 degrees C but was unaffected at pH 7.4 at the same temperature. The cell wall was composed of two nonglycosylated polypeptides (114 and 110 kDa). The cell wall resembled an archaeal S layer and may function in regulating the passage of small (< 10-kDa) sheath precursor proteins. PMID- 8244925 TI - Analysis of Streptococcus pyogenes promoters by using novel Tn916-based shuttle vectors for the construction of transcriptional fusions to chloramphenicol acetyltransferase. AB - We have developed a series of shuttle vectors based on the conjugative transposon Tn916 that have been designed for the analysis of transcriptional regulation in Streptococcus pyogenes and other gram-positive bacteria. Designated the pVIT vectors (vectors for integration into Tn916), the vectors are small, stable plasmids in Escherichia coli to facilitate the fusion of promoters from cloned S. pyogenes genes to a promoterless gene which encodes chloramphenicol acetyltransferase. The vectors each contain one or more small regions of Tn916 to direct the integration of the transcriptional fusion into the transposon via homologous recombination following transformation of S. pyogenes or other suitable gram-positive hosts. Integration can be monitored by the inactivation or replacement of an antibiotic resistance determinant in modified derivatives of Tn916. Promoter activity can then be quantitated by the determination of chloramphenicol acetyltransferase-specific activity. In addition, since integration is into loci that do not disrupt the conjugative transpositional functions of Tn916, the vectors are useful for analysis of regulation in strains that are difficult or impossible to transform and can be introduced into these strains by conjugation following transformation of an intermediate host. The promoters for the genes which encode both the M protein and protein F of S. pyogenes were active in pVIT vectors, as was the region which controls transcription of mry, a trans-acting positive regulator of M protein expression. However, neither of the two characterized promoters for mry demonstrated activity when independently analyzed in pVIT-generated partial diploid strains, suggesting that regulation of mry is more complex than predicted by current models. The broad host range of Tn916 should make the pVIT vectors useful for analysis of regulation in numerous other bacterial species. PMID- 8244926 TI - The tcmVI region of the tetracenomycin C biosynthetic gene cluster of Streptomyces glaucescens encodes the tetracenomycin F1 monooxygenase, tetracenomycin F2 cyclase, and, most likely, a second cyclase. AB - Certain mutations in the tcmVI region of the Streptomyces glaucescens chromosome affect formation of the D ring of the polyketide antibiotic tetracenomycin C (TCM C). This region lies immediately upstream from the TCM C polyketide synthase genes (tcmKLM), and the nucleotide sequence reveals the presence of three small genes, tcmH, tcmI, and tcmJ. On the basis of the phenotypes of mutants and the effects of these genes, when coupled on a plasmid with the tcmKLMN177 genes (tcmN177 is a 3'-truncated version of tcmN), on the production of TCM intermediates in a TCM- mutant, the tcmH gene encodes the C-5 monooxygenase that converts TCM F1 to TCM D3, the tcmI gene encodes the D-ring cyclase that converts TCM F2 to TCM F1 (mutations in this gene are responsible for the type VI phenotype), and the tcmJ gene most likely encodes the B-ring cyclase that acts in the biosynthesis of TCM F2. Furthermore, it appears that the N-terminal domain of the tcmN gene product (encoded by the tcmN177 gene) acts later in the biosynthesis of TCM F2 than the product of tcmJ, suggesting that the N-terminal domain of the TcmN protein is the C-ring cyclase. PMID- 8244927 TI - Regions of the Bacillus subtilis ilv-leu operon involved in regulation by leucine. AB - The ilv-leu operon of Bacillus subtilis is regulated in part by transcription attenuation. The cis-acting elements required for regulation by leucine lie within a 683-bp fragment of DNA from the region upstream of ilvB, the first gene of the operon. This fragment contains the ilv-leu promoter and 482 bp of the ilv leu leader region. Spontaneous mutations that lead to increased expression of the operon were shown to lie in an imperfect inverted repeat encoding the terminator stem within the leader region. Mutations within the inverted repeat of the terminator destroyed most of the leucine-mediated repression. The remaining leucine-mediated repression probably resulted from a decrease in transcription initiation. A systematic analysis of other deletions within the ilv-leu leader region identified a 40-bp region required for the derepression that occurred during leucine limitation. This region lies within a potential RNA stem-and-loop structure that is probably required for leucine-dependent control. Deletion analysis also suggested that alternate secondary structures proximal to the terminator are involved in allowing transcription to proceed beyond the terminator. Additional experiments suggested that attenuation of the ilv-leu operon is not dependent on coupling translation to transcription of the leader region. Our data support a model proposed by Grundy and Henkin (F. J. Grundy and T. M. Henkin, Cell 74:475-482, 1993) in which uncharged tRNA acts as a positive regulatory factor to increase gene expression during amino acid limitation. PMID- 8244928 TI - Genetic analysis of the marine manganese-oxidizing Bacillus sp. strain SG-1: protoplast transformation, Tn917 mutagenesis, and identification of chromosomal loci involved in manganese oxidation. AB - Mature spores of the marine Bacillus sp. strain SG-1 bind and oxidize manganese(II), thereby becoming encrusted with a manganese(IV) oxide. Both the function and mechanism of this oxidation are unknown, although evidence suggests that spore coat proteins are involved. To further study this phenomenon, methods of genetic analysis were developed for SG-1. By a modified protoplast transformation procedure, SG-1 was transformed (approximately 100 transformants per micrograms of DNA) with several different plasmids of gram-positive origin. Transposon Tn917, delivered on the temperature-sensitive plasmid pLTV1, was used to generate mutants of SG-1. Conditions were established that allowed 98% plasmid loss and insertions to be recovered at a frequency of 10(-3). Each mutant was found to be the result of a single insertion event. Restriction analysis of 27 mutants that do not oxidize manganese but still sporulate localized 17 of the insertions within two regions of the chromosome (termed Mnx regions), and a physical map of these regions was generated. Analysis of 18 transposon integrants in which manganese oxidation was unaffected revealed random transposon integration, with none of their insertions mapping within the Mnx regions. The Mnx regions were cloned from wild-type SG-1, and the largest region, carried on the lactococcal plasmid pGK13, was used to complement in trans one of the nonoxidizing mutants. These results demonstrate that the Mnx regions encode factors that are required for the oxidation of manganese, and this represents the first report identifying genes involved in bacterial manganese oxidation. PMID- 8244929 TI - Cloning and sequencing of the cell division gene pbpB, which encodes penicillin binding protein 2B in Bacillus subtilis. AB - The pbpB gene, which encodes penicillin-binding protein (PBP) 2B of Bacillus subtilis, has been cloned, sequenced, mapped, and mutagenized. The sequence of PBP 2B places it among the class B high-molecular-weight PBPs. It appears to contain three functional domains: an N-terminal domain homologous to the corresponding domain of other class B PBPs, a penicillin-binding domain, and a lengthy carboxy extension. The PBP has a noncleaved signal sequence at its N terminus that presumably serves as its anchor in the cell membrane. Previous studies led to the hypothesis that PBP 2B is required for both vegetative cell division and sporulation septation. Its sequence, map site, and mutant phenotype support this hypothesis. PBP 2B is homologous to PBP 3, the cell division protein encoded by pbpB of Escherichia coli. Moreover, both pbpB genes are located in the same relative position within a cluster of cell division and cell wall genes on their respective chromosomes. However, immediately adjacent to the B. subtilis pbpB gene is spoVD, which appears to be a sporulation-specific homolog of pbpB. Inactivation of SpoVD blocked synthesis of the cortical peptidoglycan in the spore, whereas carboxy truncation of PBP 2B caused cells to grow as filaments. Thus, it appears that a gene duplication has occurred in B. subtilis and that one PBP has evolved to serve a common role in septation during both vegetative growth and sporulation, whereas the other PBP serves a specialized role in sporulation. PMID- 8244930 TI - An extraintestinal, pathogenic isolate of Escherichia coli (O4/K54/H5) can produce a group 1 capsule which is divergently regulated from its constitutively produced group 2, K54 capsular polysaccharide. AB - We are studying an O4/K54/H5 Escherichia coli bacteremic isolate (CP9) as a model pathogen for extraintestinal infection. Its group 2, K54 capsular polysaccharide is an important virulence determinant and confers serum resistance. In this study the effect of the group 1 capsule regulators, RcsA, RcsB, and Lon protease, on the regulation of CP9's capsular polysaccharides was assessed. It was established that in the presence of multicopy rcsA or with disruption of lon, CP9 can be induced to produce a group 1 capsule. RcsA, RcsB, and Lon are present in this K54 background and regulate group 1 capsule expression in a fashion similar to that described for K-12 strains. Two independent group 2 capsule gene protein fusions (cl1.29::TnphoA and cl1.137::TnphoA) were used to evaluate the effects of these regulators on group 2 K54 capsule production. Disruption of lon resulted in 1.9 fold (TR293 [cl1.29::TnphoA lon-146]) and 3.4-fold (TR1373 [cl1.137::TnphoA lon 146]) decreases in fusion activity at 28 degrees C, relative to the baseline level. However, decreases in fusion activity at 42 degrees C were only 1.2- and 1.4-fold, respectively. Inactivation of both lon and rcsA or lon and rcsB restored fusion activity to baseline levels at 28 degrees C, but only a partial restoration of activity was seen at higher temperatures. To assess whether these differences in fusion activity reflected a functional change in capsule production, the effects of 80% normal human serum (NHS) were tested against CP9 and TR93 (lon-146). Since the group 2 K54 capsule protects against the bactericidal activity of 80% NHS, a decrease in its production results in an increase in serum sensitivity. Viable counts of CP9 increased 10-fold in 80% NHS over 3 h at 28 degrees C, as expected. In contrast to CP9, TR93 (lon-146) incurred a 10-fold loss in viability under the same conditions. The levels of RcsA are increased in TR93 (lon 146) as consequence of lon disruption; therefore, these results in conjunction with the cl1::TnphoA protein fusion data establish RcsA as a negative regulator of the group 2 K54 capsular polysaccharide. Furthermore, these results also suggest existence of another Lon-sensitive negative regulator of group 2 K54 capsule production, which is active higher temperatures. PMID- 8244931 TI - Location of IS200 on the genomic cleavage map of Salmonella typhimurium LT2. AB - Locations of six Tn10s, closely linked to each of the six IS200s on the genomic cleavage map of Salmonella typhimurium LT2, were determined by digestion with XbaI and BlnI and separation of the fragments by pulsed-field gel electrophoresis; the locations were then further defined by P22-mediated joint transduction. The orientation of each IS200 with respect to its linked Tn10 was determined by Southern blotting. The locations of IS200-I, IS200-III, and IS200-V were confirmed to be close to sufD, melB, and purC, as previously indicated. IS200-II is jointly transduced with cysG. IS200-IV is near fliA; the linked Tn10 is inserted in fli, making the strain nonmotile. IS200-VI is jointly transduced with aspC but not with aroA. IS200 is transposed to a seventh site in some strains, while remaining in the other six locations described above. These data indicate that genome analysis by pulsed-field gel electrophoresis can locate the positions of Tn10s with accuracy sufficient to predict P22-mediated joint transduction. PMID- 8244932 TI - Transcription properties of RNA polymerase holoenzymes isolated from the purple nonsulfur bacterium Rhodobacter sphaeroides. AB - We have been characterizing RNA polymerase holoenzymes from Rhodobacter sphaeroides. RNA polymerase purified from R. sphaeroides transcribed from promoters recognized by Escherichia coli E sigma 32 or E sigma 70 holoenzyme. Antisera to E. coli sigma 32 or sigma 70 indicated that related polypeptides of approximately 37 kDa (sigma 37) and 93 kDa (sigma 93), respectively, are present in this preparation. Transcription of sigma 32-dependent promoters was observed in a further fractionated R. sphaeroides holoenzyme containing the sigma 37 polypeptide, while a preparation enriched in sigma 93 transcribed sigma 70 dependent promoters. To demonstrate further that the sigma 93 polypeptide functions like E. coli sigma 70, we obtained an R. sphaeroides E sigma 93 holoenzyme capable of transcription from sigma 70-dependent promoters by combining sigma 93 with (i) an E sigma 37 fraction with diminished sigma 93 polypeptide content or (ii) E. coli core RNA polymerase. The generation of analogous DNase I footprints on the lacUV5 promoter by R. sphaeroides E sigma 93 and by E. coli E sigma 70 suggests that the overall structures of these two holoenzymes are similar. However, some differences in promoter specificity between R. sphaeroides E sigma 93 and E. coli E sigma 70 exist because transcription of an R. sphaeroides rRNA promoter was detected only with E sigma 93. PMID- 8244933 TI - Insertional inactivation of genes to isolate mutants of Synechococcus sp. strain PCC 7942: isolation of filamentous strains. AB - We have developed a simple procedure for generating mutants of the cyanobacterium Synechococcus sp. strain PCC 7942 in which the site of the lesion can be readily identified. This procedure involves transforming Synechococcus sp. strain PCC 7942 with a library of its own DNA that was fully digested with Sau3A and ligated into the plasmid vector pUC8. The homologous integration of the recombinant plasmid into the genome will often result in the disruption of a gene and the loss of gene function. We have used this method to generate many mutants of Synechococcus sp. strain PCC 7942 which grow as multicellular filaments rather than as unicells. Since the gene harboring the lesion was tagged with pUC8, it was easily isolated. In this paper, we discuss the usefulness of this procedure for the generation of mutants, and we characterize one mutant in which the lesion may be in an operon involved in the assembly of lipopolysaccharides. PMID- 8244934 TI - Identification of the Shiga toxin A-subunit residues required for holotoxin assembly. AB - Recent X-ray crystallographic analyses have demonstrated that the receptor binding (B) subunits of Shiga toxin (STX) are arranged as a doughnut-shaped pentamer. The C terminus of the enzymatic (A) subunit presumably penetrates the nonpolar pore of the STX B pentamer, and the holotoxin is stabilized by noncovalent interactions between the polypeptides. We identified a stretch of nine nonpolar amino acids near the C terminus of StxA which were required for subunit association by using site-directed mutagenesis to introduce progressive C terminal deletions in the polypeptide and assessing holotoxin formation by a receptor analog enzyme-linked immunosorbent assay, immunoprecipitation, and a cytotoxicity assay. Tryptophan and aspartic acid residues which form the N terminal boundary, as well as two arginine residues which form the C-terminal boundary of the nine-amino-acid sequence, were implicated as the stabilizers of subunit association. Our model proposes that residues 279 to 287 of the 293-amino acid STX A subunit penetrate the pore while the tryptophan, aspartic acid, and 2 arginine residues interact with other charged or aromatic amino acids outside the pore on the planar surfaces of the STX B pentamer. PMID- 8244935 TI - Cloning and characterization of the Pseudomonas aeruginosa sodA and sodB genes encoding manganese- and iron-cofactored superoxide dismutase: demonstration of increased manganese superoxide dismutase activity in alginate-producing bacteria. AB - Pseudomonas aeruginosa is a strict aerobe which is likely exposed to oxygen reduction products including superoxide and hydrogen peroxide during the metabolism of molecular oxygen. To counterbalance the potentially hazardous effects of elevated endogenous levels of superoxide, most aerobic organisms possess one or more superoxide dismutases or compounds capable of scavenging superoxide. We have previously shown that P. aeruginosa possesses both an iron- and a manganese-cofactored superoxide dismutase (D. J. Hassett, L. Charniga, K. A. Bean, D. E. Ohman, and M. S. Cohen, Infect. Immun. 60:328-336, 1992). In this study, the genes encoding manganese (sodA)- and iron (sodB)- cofactored superoxide dismutase were cloned by using a cosmid library of P. aeruginosa FRD which complemented an Escherichia coli (JI132) strain devoid of superoxide dismutase activity. The sodA and sodB genes of P. aeruginosa, when cloned into a high-copy-number vector (pKS-), partially restored the aerobic growth rate defect, characteristic of the Sod- strain, to that of the wild type (AB1157) when grown in Luria broth. The nucleotide sequences of sodA and sodB have open reading frames of 612 and 579 bp that encode dimeric proteins of 22.9 and 21.2 kDa, respectively. These data were also supported by the results of in vitro expression studies. The deduced amino acid sequence of the P. aeruginosa manganese and iron superoxide dismutase revealed approximately 50 and 67% similarity with manganese and iron superoxide dismutases from E. coli, respectively. There was also remarkable similarity with iron and manganese superoxide dismutases from other phyla. The mRNA start site of sodB was mapped to 174 bp upstream of the ATG codon. A likely promoter with similarity to the -10 and -35 consensus sequence of E. coli was observed upstream of the ATG start codon of sodB. Regions sequenced 519 bp upstream of the sodA electrophoresis, sodA gene revealed no such promoter, suggesting an alternative mode of control for sodA. By transverse field electrophoresis, sodA and sodB were mapped to the 71- to 75-min region on the P. aeruginosa PAO1 chromosome. Strikingly, mucoid alginate-producing bacteria generated greater levels of manganese superoxide dismutase than nonmucoid revertants, suggesting that mucoid P. aeruginosa is responding to oxidative stress and/or changes in the redox status of the cell. PMID- 8244936 TI - The xynC gene from Fibrobacter succinogenes S85 codes for a xylanase with two similar catalytic domains. AB - The xynC gene of Fibrobacter succinogenes S85 codes for a 66.4-kDa xylanase which consists of three distinct domains separated by two flexible regions rich in serine residues. Domains A and B of XynC code for catalytic domains with 56.5% identity and 9.6% similarity with each other, and both domains share homology with xylanases of Ruminococcus flavefaciens, Neocallimastix patriciarum, Clostridium acetobutylicum, Bacillus pumilus, Bacillus subtilis, and Bacillus circulans. More than 88% of the xylanase activity of Escherichia coli cells carrying the original 13-kb recombinant plasmid was released from intact cells by cold water washes. The major products of hydrolysis of xylan by both domains were xylose and xylobiose, indicating that the xynC gene product exhibits catalytic properties similar to those of the XynA xylanases from R. flavefaciens and N. patriciarum. So far, these features are not shared broadly with bacteria from other environments and may indicate specific selection for this domain structure in the highly competitive environment of the rumen. PMID- 8244937 TI - Genetic and molecular analyses of the C-terminal region of the recE gene from the Rac prophage of Escherichia coli K-12 reveal the recT gene. AB - The nucleotide sequence of the C-terminal region of the recE gene of the Rac prophage of Escherichia coli K-12 reveals the presence of a partially overlapping reading frame we call recT. Deletion mutations show that recT is required for the RecE pathway of conjugational recombination. By cloning recT with a plasmid vector compatible with pBR322, we showed by cis-trans tests that the portion of the recE gene encoding ExoVIII DNA nuclease activity is also required for RecE pathway conjugational recombination. The recT gene can replace the redB gene of lambda for recA-independent plasmid recombination. A Tn10 insertion mutation previously thought to be in recE is located in recT and is renamed recT101::Tn10. Discrepancies between the molecular mass estimates of wild-type ExoVIII protein determined from mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and calculated from the predicted amino acid sequence are discussed. The hypothesis that wild-type ExoVIII protein results from fusion of RecE and RecT proteins is disproved genetically, thus supporting a previous hypothesis that the discrepancies are due to abnormal protein mobility in SDS PAGE. A computer-performed scan of the bacteriophage nucleotide sequence data base of GenBank revealed substantial similarity between most of recE and a 2.5-kb portion of the b2 region of lambda. This suggests interesting speculations concerning the evolutionary relationship of lambda and Rac prophages. PMID- 8244938 TI - In vitro activity of NifL, a signal transduction protein for biological nitrogen fixation. AB - In the free-living diazotroph Klebsiella pneumoniae, the NifA protein is required for transcription of all nif (nitrogen fixation) operons except the regulatory nifLA operon itself. NifA activates transcription of nif operons by the alternative holoenzyme form of RNA polymerase, sigma 54 holoenzyme. In vivo, NifL is known to antagonize the action of NifA in the presence of molecular oxygen or combined nitrogen. We now demonstrate inhibition by NifL in vitro in both a coupled transcription-translation system and a purified transcription system. Crude cell extracts containing NifL inhibit NifA activity in the coupled system, as does NifL that has been solubilized with urea and allowed to refold. Inhibition is specific to NifA in that it does not affect activation by NtrC, a transcriptional activator homologous to NifA, or transcription by sigma 70 holoenzyme. Renatured NifL also inhibits transcriptional activation by a maltose binding protein fusion to NifA in a purified transcription system, indicating that no protein factor other than NifL is required. Since inhibition in the purified system persists anaerobically, our NifL preparation does not sense molecular oxygen directly. PMID- 8244939 TI - Physiological characterization of putative high-affinity glucose transport protein Hxt2 of Saccharomyces cerevisiae by use of anti-synthetic peptide antibodies. AB - Characterization and quantification of the Hxt2 (hexose transport) protein of Saccharomyces cerevisiae indicate that it is one of a set of differentially expressed high-affinity glucose transporters. The protein product of the HXT2 gene was specifically detected by antibodies raised against a synthetic peptide encompassing the 13 carboxyl-terminal amino acids predicted by the HXT2 gene sequence. Hxt2 migrated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a broad band or closely spaced doublet with an average M(r) of 47,000. Hxt2 cofractionated with the plasma membrane ATPase, Pma1, indicating that it is a plasma membrane protein. Hxt2 was not solubilized by high pH or urea but was solublized by detergents, which is characteristic of an integral membrane protein. Expression of the Hxt2 protein was measured under two different conditions that produce expression of high-affinity glucose transport: a medium shift from a high (2.0%) to a low (0.05%) glucose concentration (referred to below as high and low glucose) and growth from high to low glucose. Hxt2 as measured by immunoblotting increased 20-fold upon a shift from high-glucose to low-glucose medium, and the high-affinity glucose transport expressed had a strong HXT2-dependent component. Surprisingly, Hxt2 was not detectable when S. cerevisiae growing in high glucose approached glucose exhaustion, and the high affinity glucose transport expressed under these conditions did not have an HXT2 dependent component. The role of Hxt2 in growth during aerobic batch culture in low-glucose medium was examined. An hxt2 null mutant grew and consumed glucose significantly more slowly than the wild type, and this phenotype correlated directly with appearance of the Hxt2 protein. PMID- 8244940 TI - Isolation and location on the R27 map of two replicons and an incompatibility determinant specific for IncHI1 plasmids. AB - Two replicons were isolated independently from different IncHI1 plasmids. One was isolated from R27, and a second was isolated from pIP522. We demonstrate, by DNA DNA hybridization experiments, that these maintenance regions are different and that they are specific to, and carried by, all IncHI1 plasmids tested. In view of this specificity we decided to designate the replicon isolated from R27 as RepHI1A and the replicon isolated from pIP522 as RepHI1B. These two autoreplicative regions are not related to a third replicon present in all IncHI1 plasmids that bears homology with RepFIA and that expresses the characteristic incompatibility of IncHI1 subgroup plasmids toward F factor (D. Saul, D. Lane, and P. L. Bergquist, Mol. Microbiol. 2:219-225, 1988; D. E. Taylor, R. W. Hedges, and P. L. Bergquist, J. Gen. Microbiol. 131:1523-1530, 1985). These results demonstrate that all IncHI1 plasmids tested contain at least three replicons. An incompatibility (Inc) region that hybridizes specifically to all the IncHI1 plasmids was previously isolated (M. Couturier, F. Bex, P. L. Bergquist, and W. K. Maas, Microbiol. Rev. 52:375-395, 1988). Although this Inc locus is not located in an autoreplicative region of IncHI1 plasmids, we observed that this locus stabilizes a low-copy-number replicon. This Inc locus is probably a component of an active partition locus involved in the maintenance of IncHI1 plasmids. The nucleotide sequence of the Inc region contains direct repeats of 31 bp. In addition, this incompatibility determinant hybridizes specifically with IncHI1 plasmids but expresses incompatibility toward plasmids of both IncHI subgroups (IncHI1 and IncHI2). In this communication, we present the mapping of these maintenance elements on the R27 genome. PMID- 8244941 TI - Biosynthesis of biotin from dethiobiotin by the biotin auxotroph Lactobacillus plantarum. AB - Lactobacillus plantarum requires biotin for growth. We show that in the presence of high levels of the biotin biosynthetic precursor, dethiobiotin, L. plantarum synthesizes biotin and grows in medium with dethiobiotin but without biotin. Lactobacillus casei also grew under similar conditions. PMID- 8244942 TI - Azotobacter vinelandii mutS: nucleotide sequence and mutant analysis. AB - An Azotobacter vinelandii homolog to the Salmonella typhimurium mutS gene was discovered upstream of the fdxA gene. The product of this gene is much more similar to S. typhimurium MutS than either is to the HexA protein of Streptococcus pneumoniae. An A. vinelandii delta mutS mutant strain was shown to have a spontaneous mutation frequency 65-fold greater than that of the wild type. PMID- 8244943 TI - The pss and psd genes are required for motility and chemotaxis in Escherichia coli. AB - Mutants of Escherichia coli defective in phosphatidylserine synthase (encoded by pss) and phosphatidylserine decarboxylase (encoded by psd) make cell membranes deficient in phosphatidylethanolamine. In this report we show that wild-type pss and psd genes are required for motility and chemotaxis. Null mutants or strains with temperature-sensitive pss or psd mutations grown at high temperature (35 degrees C) were nonmotile. They lacked flagella and showed reduced rates of transcription of the flhD master operon (encoding FlhD and FlhC), the fliA operon (encoding sigma F), and the fliC operon (encoding flagellin). At low temperature (25 degrees C), the temperature-sensitive mutant cells showed motility and chemotaxis but at reduced levels. The extent of the motility and chemotaxis defects in the mutants was correlated with the amount of phosphatidylethanolamine in the membranes, suggesting a link between membrane phospholipid composition and expression of the flagellum chemotaxis regulon. PMID- 8244944 TI - Altered-function mutations in the Agrobacterium tumefaciens OccR protein and in an OccR-regulated promoter. AB - OccR is a LysR-type transcriptional activator that controls the occQ and traR promoters of octopine-type Ti plasmids. The opine octopine converts OccR from a repressor to an activator of occQ, shortens the protein's DNase I footprint, and decreases the angle of an OccR-caused DNA bend at the occQ promoter. In this study we first localized the cis-acting DNA sequences required for regulated expression of occQ. To understand better the mechanism of activation of OccR, we isolated mutations both in the occQ promoter and in the occR gene which function differently from the wild type. An occQ promoter mutation that changes the putative -35 region of occQ from TTGACC to TTGACA increases the basal expression of occQ about 15-fold. Three mutations in occR were also identified, one of which activates occQ at fully constitutive levels in both the absence and presence of octopine. This mutation (E23G) is located in the first helix of a putative helix turn-helix DNA-binding motif. The other two occR mutations cause the protein to detect much lower concentrations of octopine than wild-type OccR protein does. These mutations (F113L and G148D) are located in a region of the protein that is predicted to contain the ligand-binding site. PMID- 8244945 TI - Energy-dependent degradation of lambda O protein in Escherichia coli. AB - Protein O of bacteriophage lambda is a short-lived protein which has a key role in the replication of the phage DNA in Escherichia coli. Here we present evidence that lambda O degradation is energy dependent: it is impaired by cyanide and alpha-methylglucoside, both of which inhibit cellular energy metabolism. Removal of these inhibitors restored the degradation of lambda O. Our experiments suggest that limited amounts of cellular energy are sufficient to support lambda O degradation. In addition, degradation of lambda O protein is prevented by a mutation in the E. coli clpP gene, but not by a mutation in the clpA gene. These results suggest that the ClpP protease is involved in the energy-dependent degradation of the lambda O protein. PMID- 8244946 TI - Mutational analysis of the bacteriophage P2 Ogr protein: truncation of the carboxy terminus. AB - The Ogr protein is a 72-residue, zinc-binding transcription factor essential for activation of late gene expression in bacteriophage P2. Analysis of C-terminal truncated proteins generated by stop codon mutagenesis shows that deletion of residues distal to position 51 had negligible effects on Ogr function. More extensive deletion resulted in unstable products with severely reduced activity. These results, as well as the effects of other mutations in this region, support the idea that the 21 C-terminal residues are not required for transactivation. PMID- 8244947 TI - Patch length of localized repair events: role of DNA polymerase I in mutY dependent mismatch repair. AB - In vivo experiments with heteroduplex lambda genomes show that the MutY mismatch repair system of Escherichia coli defines an average repair tract that is shorter than 27 nucleotides and longer than 9 nucleotides and extends 3' from the corrected adenine. The phenotype of a mutant defective in DNA polymerase I shows that this enzyme plays a significant, though not an essential, role in the in vivo repair of apurinic sites generated by this system. Evidence is presented that in the absence of polymerase I the repair tracts are modestly longer than in the polA+ extending in the 5' direction from the corrected adenine, suggesting a role for another DNA polymerase. PMID- 8244948 TI - Evidence that peptide deformylase and methionyl-tRNA(fMet) formyltransferase are encoded within the same operon in Escherichia coli. AB - Overexpression of the fms gene, the first translation unit of a dicistronic operon that also encodes methionyl-tRNA(fMet) formyltransferase in Escherichia coli, sustains the overproduction of peptide deformylase activity in crude extracts. This suggests that the fms gene encodes the peptide deformylase. Moreover, the fms gene product has a motif characteristic of metalloproteases, an activity compatible with deformylase. The corresponding protein could be purified to homogeneity. However, its enzymatic activity could not be retained during the purification procedure. As could be expected from the occurrence in its amino acid sequence of a zinc-binding motif characteristic of metallopeptidases, the purified fms product displayed one tightly bound zinc atom. PMID- 8244949 TI - The FhuA protein is involved in microcin 25 uptake. AB - A chromosomal Tn5 insertion resulting in complete resistance to the peptide antibiotic microcin 25 was mapped to the min 4 region of the Escherichia coli genetic map. Additional experiments showed that the insertion disrupted the fhuA gene, which encodes the multifunctional outer membrane receptor for ferrichrome, the antibiotic albomycin, colicin M, and bacteriophages T5, T1, and phi 80. Thus, microcin 25 and all of these agents share the same receptor. PMID- 8244950 TI - The dihydropteroate synthase gene, folP, is near the leucine tRNA gene, leuU, on the Escherichia coli chromosome. PMID- 8244951 TI - Physical mapping of the Escherichia coli pepT and potABCD genes. PMID- 8244952 TI - Physical map location of a set of Escherichia coli genes (hde) whose expression is affected by the nucleoid protein H-NS. PMID- 8244953 TI - Location of the hemE gene on the physical map of Escherichia coli. PMID- 8244954 TI - Genetic map of the tyrT region of Escherichia coli from 27.1 to 27.7 minutes based exclusively on sequence data. PMID- 8244955 TI - Yeast DNA replication. PMID- 8244956 TI - Targeting of Bcl-2 to the mitochondrial outer membrane by a COOH-terminal signal anchor sequence. AB - The protooncogene product Bcl-2 is an integral membrane protein that functions as a suppressor of programmed cell death. It contains a single predicted transmembrane segment located at its COOH terminus. Here, we show that the transmembrane domain of human Bcl-2 functions as a mitochondrial signal anchor sequence that targets and inserts the protein into the outer membrane in an Ncyto C(in) orientation, leaving the bulk of the polypeptide facing the cytosol. Deletion of the COOH-terminal 22 amino acids of Bcl-2 abrogated protein targeting, whereas fusion of this domain to the COOH terminus of dihydrofolate reductase resulted in targeting and insertion of the hybrid protein into the outer membrane in a manner similar to that of Bcl-2. The sequence of the hydrophobic core of the Bcl-2 signal anchor is similar to the corresponding region of the NH2-terminal signal anchor of the mitochondrial outer membrane protein in yeast, Mas70p. A synthetic peptide comprising the Mas70p signal anchor sequence effectively competed for insertion of Bcl-2 into the outer membrane but had no effect on the comparatively low association that Bcl-2 makes with endoplasmic reticulum microsomes. Insertion of Bcl-2 into the mitochondrial outer membrane is mechanistically different than its association with microsomes. PMID- 8244957 TI - Integrin alpha 7 as substrate for a glycosylphosphatidylinositol-anchored ADP ribosyltransferase on the surface of skeletal muscle cells. AB - An arginine-specific mono-ADP-ribosyltransferase is expressed on the surface of differentiated mouse skeletal muscle cells and is anchored in the membrane via a glycosylphosphatidylinositol tail. Following incubation of intact cells with [adenylate-32P]NAD and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), a 97-kDa [32P]ADP-ribosylated protein was observed under reducing conditions and a 140-kDa complex under nonreducing conditions. The ADP-ribosylated protein was purified on a laminin affinity column. Based on its N terminal sequence (FNLDVM-GAIRKEGEPGSLFGF) and a partial internal sequence (GLMRSEELSFVAGAP), the modified protein was identified as integrin alpha 7. Following partial trypsin digestion, a 39-kDa/79-kDa radiolabeled fragment was produced (reduced/nonreduced SDS-PAGE), narrowing the ADP-ribosylation site to a 39-kDa segment in the extracellular domain of integrin alpha 7. Labeling under optimal conditions was at least 0.4 mol of ADP-ribose/mol of integrin alpha 7. Selective expression of both ADP-ribosyltransferase and integrin alpha 7 in cardiac and skeletal muscle, a similar developmental appearance, and the apparently specific ADP-ribosylation, are consistent with a regulatory association between these proteins. ADP-ribosylation may modulate integrin receptor signaling and could play a significant role in the regulation of muscle cell function by the extracellular matrix. PMID- 8244958 TI - Regulation of rDNA transcription by insulin in primary cultures of rat hepatocytes. AB - The studies presented herein were designed to investigate the role of insulin in maintaining the steady-state number of ribosomes in primary cultures of rat hepatocytes. The RNA content of hepatocytes maintained in the presence of insulin did not change during the 7 days of culture. In contrast, the RNA content declined significantly following 2 days of insulin deprivation and after 4 days without insulin reached a new steady-state value that was approximately 60% of that observed for hepatocytes maintained in the presence of the hormone. Following addition of insulin, the RNA content of insulin-deprived hepatocytes started to increase within 6 h and was restored to the control value within 48 h. The amounts of 18 and 28 S RNA, and thus the number of ribosomes, changed in concert with the total RNA content. Furthermore, synthesis of total cellular protein responded in parallel to the changes in RNA content, suggesting that the number of ribosomes was the primary determinant of protein synthesis under the conditions of these experiments. About one-half of the increase in RNA content following the addition of insulin to insulin-deprived cells could be accounted for by a reduction in the rate of degradation of ribosomes. The remainder of the change in RNA content must have resulted from an increase in ribosome biogenesis. This possibility was confirmed by measuring [3H]uridine incorporation into ribosomal RNA present in mature ribosomes. The rate of synthesis of ribosomal RNA was reduced in parallel with the fall in RNA content in insulin-deprived hepatocytes and was restored to the control value within 3 h of the addition of insulin. Alterations in the synthesis of ribosomal RNA in response to insulin deprivation and replacement were associated with parallel changes in transcription of rDNA as measured in nuclear run-on assays using a DNA probe corresponding to the external transcribed spacer region of rat rDNA. The data demonstrate that insulin regulates the number of ribosomes in primary cultures of rat hepatocytes by accelerating the rate of transcription of rDNA and by slowing the rate of ribosome degradation. PMID- 8244959 TI - Engineered topographic determinants with alpha beta, beta alpha beta, and beta alpha beta alpha topologies show high affinity binding to native protein antigen (lactate dehydrogenase-C4). AB - The use of peptides has attracted much interest in the development of synthetic vaccines. Although our current understanding of peptide antigens as immunogens has been greatly advanced recently, there still remain many obstacles. The B cell response elicited by a peptide antigen is governed by a number of poorly understood events such as epitope structure, T cell dependency and major histocompatibility complex restriction, adjuvancy, route of immunization, and immunogen stability. In this paper, we extend our previous studies on the problem of the topographical nature of antigenic sites on native protein antigens, in terms of how much molecular mimicry must be maintained in an antigenic determinant for the induction of high affinity antibodies specific for native protein. We show here that an antigenic epitope from the model contraceptive vaccine candidate lactate dehydrogenase (LDH-C4) can be rationally engineered into a highly structured conformation that mimics the corresponding site in the native three-dimensional protein. Our strategy is based on the selection of an antigenic segment which exhibits certain secondary structural properties and by design principles is fixed in three dimensions by appropriate grafting onto a supersecondary structural motif such as alpha beta, beta alpha beta, or beta alpha beta alpha. The biophysical data are consistent with the proposed secondary structures, and antibodies raised to the various construct show high affinity for the native protein. These studies lend further credence to the conformational nature of peptide epitopes and provide a basis for the rational design of peptide vaccines. PMID- 8244960 TI - The interplay of the GrpE heat shock protein and Mg2+ in RepA monomerization by DnaJ and DnaK. AB - Genetic and biochemical studies have established that the sole function of the Escherichia coli DnaJ, DnaK, and GrpE heat shock proteins in plasmid P1 DNA replication is to convert RepA dimers to monomers. Monomers bind avidly to oriP1 DNA and initiate DNA replication. However, with purified heat shock proteins, only DnaJ, DnaK, and ATP were required for the monomerization of RepA; GrpE was not required. We have found reaction conditions that mimic the physiological situation. GrpE function is absolutely necessary for RepA activation in vitro with DnaJ and DnaK when the free Mg2+ concentration is maintained at a level of approximately 1 microM by a metal ion buffer system. EDTA or physiological metabolites, including citrate, phosphate, pyrophosphate, and ATP, all elicit the GrpE requirement. With these metal ion-buffering systems, GrpE specifically lowers the concentration of Mg2+ required for the RepA activation reaction. The absence of Mg2+ blocks activation and high levels of Mg2+ in solution bypass the requirement for GrpE but not for the other two heat shock proteins. Our results imply that GrpE facilitates the utilization of Mg2+ for an essential step in RepA activation. PMID- 8244961 TI - A novel ecto-phosphatidic acid phosphohydrolase activity mediates activation of neutrophil superoxide generation by exogenous phosphatidic acid. AB - Phosphatidic acid (PA) added to intact cells activates a variety of processes including mitogenesis in fibroblasts and superoxide generation in neutrophils. We have investigated the mechanism of activation of superoxide generation in intact human neutrophils by a short-chain (dioctanoyl) PA (diC8PA). After a lag, diC8PA caused a high rate of superoxide production (19.6 nmol of cytochrome c reduced/min/10(6) cells). Activation did not require extracellular Ca2+ and coincided with near quantitative conversion of diC8PA to dioctanoylglycerol (diC8 glycerol). diC8PA also activated cellular phospholipase D with release of long chain PA and secondary production of long-chain diradylglycerol (sn-1,2 diacylglycerol and 1-O-alkyl-2-acylglycerol). The metabolism of diC8PA to diC8 glycerol was catalyzed by a novel PA phosphohydrolase on the outer leaflet of the plasma membrane as demonstrated by the exclusive release of Pi into the extracellular medium. This enzyme also showed activity toward PA containing long chain unsaturated fatty acids. The ecto-PA phosphohydrolase differed from the intracellular PA phosphohydrolase based on its relative insensitivity to desipramine and N-ethylmaleimide. The enzyme was also present in Chinese hamster ovary (CHO) cells and its activity did not change in transfected CHO cells expressing the two membrane-associated isoforms of alkaline phosphatase, indicating that the PA phosphohydrolase was not alkaline phosphatase. Non hydrolyzable phosphonate analogs of diC8PA poorly stimulated superoxide production. Activation of superoxide generation by diC8PA was inhibited by staurosporine, suggesting a protein kinase C-dependent mechanism. We suggest that the action of a novel ecto-PA phosphohydrolase permits exogenously added short chain PA to serve as "timed-release diacylglycerol" and that its biological effects in neutrophils are secondary to diacylglycerol-mediated protein kinase C activation. PMID- 8244962 TI - Differential acute-phase response of rat kininogen genes involves type I and type II interleukin-6 response elements. AB - The serum concentration of rat T1 kininogen increases 20-30-fold in response to acute inflammation. This increase, induced in the liver, is regulated primarily at the transcriptional level. In contrast, synthesis of a homologous K kininogen is not induced. In this study, we further analyzed a 321-base pair interleukin (IL)-6 response element in the T1 kininogen promoter and showed that it consists of at least three functionally distinct sequences (A, B, and C boxes). All three sequences were required for full promoter activity. The B box, a strong C/EBP binding site, was crucial for T1 kininogen's basal expression, whereas A and C boxes resembled the type II IL-6 response elements and were critical for the cytokine response. C/EBP alpha, -beta, and -delta interacted with the B box sequence; however, upon IL-6 stimulation, C/EBP delta binding activity was dramatically induced and became the predominant factor binding to this site. Consistent with these binding studies were the cotransfection experiments, revealing that C/EBP delta was the most potent transactivator under induced conditions and that its transactivation on the T1 kininogen promoter required an intact B box. These findings substantiated the importance of the B box in eliciting the full acute-phase response. A sequence comparison showed the K kininogen promoter contained identical A and B boxes but differed from the T1 kininogen promoter by two nucleotides at the C box. This divergence reduced the IL-6 response by approximately 4-fold, thus contributing to the differential inflammatory response. Our studies demonstrate that evolutionary divergence of a few nucleotides at a critical sequence in the promoter regions can profoundly alter the expression patterns of downstream genes. PMID- 8244963 TI - The adenine nucleotide translocase modulates oligomycin-induced quenching of pyranine fluorescence in submitochondrial particles. AB - Incorporation of the fluorescent, nonpermeant pH indicator pyranine into submitochondrial particles (pyranine-SMP) permitted monitoring of intravesicular pH changes brought about by proton translocation due to oxidation of respiratory chain substrates or to hydrolysis of ATP. Addition of oligomycin to beef heart pyranine-SMP was followed by a pH-independent quenching of pyranine fluorescence. Quenching was influenced by the presence of adenine nucleotides both inside and outside the submitochondrial particles. The nature of the nucleotides required for quenching resembled the specificity of the adenine nucleotide translocase rather than F1-ATPase. Removal of F1 from pyranine-SMP by treatment of the particles with urea did not alter oligomycin-induced quenching. Atractyloside, a specific inhibitor of the adenine nucleotide translocase, prevented oligomycin induced quenching when the inhibitor was coincorporated into submitochondrial particles with pyranine. Bongkrekic acid prevented or reversed the oligomycin dependent quenching when added to pyranine-SMP either before or after oligomycin, respectively, but only when ATP was present within the particles. A mutant of Saccharomyces cerevisiae, lacking translocase genes, exhibited oligomycin dependent fluorescence quenching which was not inhibited by bongkrekic acid. The results support the interpretation that oligomycin promotes sequestration of the fluorescent probe in a region of the submitochondrial particle, probably the F0F1 complex, that leads to a quenching of fluorescence. The observed quenching can be modulated in a way that suggests an interaction between the translocase and F0. PMID- 8244964 TI - Activation of p21ras by nerve growth factor in embryonic sensory neurons and PC12 cells. AB - p21ras is believed to be involved in the neuronal differentiation of cells responsive to nerve growth factor (NGF). We show that NGF stimulates the activation of p21ras in embryonic sensory neurons and in PC12 cells. In the initial 5 min of exposure to NGF, the activation is concentration-dependent. In the sensory neurons and PC12 cells, the apparent maximal activation was reached at 50 and 10 ng/ml, respectively, with half-maximal activation at approximately 5 and 2-3 ng/ml, respectively. Kinetic analysis at low concentrations of NGF showed that p21ras activation slowly increases with time in both types of cells, while high concentrations result in rapid activation within 5 min. These results indicate that NGF regulates the activation state of p21ras in these cells and provides evidence suggesting that activation of p21ras is involved in NGF signal transduction. Treatment of PC12 cells with brain-derived neurotrophic factor or neurotrophin-3 (NT-3) failed to activate p21ras, suggesting that binding alone to p75LNGFR is insufficient for ras activation. Treatment with the kinase inhibitor, K252a, which inhibits the NGF tyrosine kinase receptor p140trk, abolished ras activation, suggesting that p140trk is the major mediator of p21ras activation by NGF. PMID- 8244965 TI - Crystal structure of adenylosuccinate synthetase from Escherichia coli. Evidence for convergent evolution of GTP-binding domains. AB - The structure of the P2(1) crystal form of adenylosuccinate synthetase from Escherichia coli has been determined to a resolution of 2.8 A. The refined model for the enzyme gives an R factor of 0.20 and a root-mean-square deviation from expected bond lengths and angles of 0.016 A and 2.27 degrees, respectively. The dominant structural element of each monomer of the homodimer is a central beta sheet of 10 strands. The first nine strands of the sheet are mutually parallel with right-handed crossover connections between the strands. The 10th strand is antiparallel with respect to the first nine strands. In addition, the enzyme has two antiparallel beta-sheets, comprised of two strands and three strands each, 11 alpha-helices and two short 3/10-helices. The overall fold of the polypeptide chain has not been observed heretofore in any other protein structure. Residues tentatively assigned to the active site of the enzyme on the basis of chemical modification and directed mutation cluster in two separate regions. Gly12, Gly15, Gly17, Lys18, Ile19, and Lys331 lie at one end of a crevice that measures 12 A by 30 A by 12 A deep. Lys140 and Arg147 are not part of this crevice, but instead lie at the interface between monomers of the dimer. Lys140 makes a salt link with Asp231 of a monomer related by molecular symmetry and Arg147 binds to the carbonyl of the same Asp231. Superposition of the p21 ras protein (Pai, E. F., Kabsch, W., Krengel, U., Holmes, K., John, J., and Wittinghofer, A. (1989) Nature 341, 209-214) onto the synthetase reveals significant correspondences between side chains of the two proteins. Residues which interact with GTP in the p21ras protein have structurally equivalent residues in the synthetase. The GTP molecule, when transformed to the coordinate frame of the synthetase, falls into the crevice defined by studies in directed mutation. We suggest that the similarities in the GTP-binding domains of the synthetase and the p21ras protein are an example of convergent evolution of two distinct families of GTP-binding proteins. PMID- 8244966 TI - Chemotactic methyltransferase promotes adaptation to repellents in Bacillus subtilis. AB - Bacillus subtilis cheRB, which encodes the chemotactic methyltransferase, has been cloned and sequenced. CheRB is a polypeptide of 256 amino acids, with a predicted molecular mass of 28 kDa. A comparison of the predicted amino acid sequence of B. subtilis CheRB with that of Escherichia coli CheRE demonstrates that the two enzymes share 31% amino acid identity. The homology was functional in that the expression of cheBB in an E. coli cheRE null mutant made the bacteria Che+. In contrast to cheRE null mutants which show a strong smooth swimming bias, cheRB null mutants were predominantly tumbly. They respond to the addition and subsequent removal of attractant. They also respond to the addition of repellent but do not adapt; they resume prestimulus bias on removal of repellent. Tethering analysis of a culture of a cheRB null mutant revealed two distinct subpopulations, each demonstrating unique behaviors. One showed a strong clockwise flagellar rotation bias, whereas the other was more random. The latter phenotype may be due to a deficiency of CheB and may reflect an interaction of CheB and CheR. Measurements of CheB activity in the cheR null mutant showed them to be only 20% of wild type levels. We conclude from this work that CheRB functions to promote adaptation to repellent stimuli in B. subtilis, whereas CheRE functions to promote adaptation to attractant stimuli in E. coli. PMID- 8244967 TI - A Chinese hamster ovary cell line with a temperature-conditional defect in receptor recycling is pleiotropically defective in lysosome biogenesis. AB - We have previously described the isolation of a Chinese hamster ovary cell line, TfT1.11, that has a pleiotropic, temperature-conditional defect in receptor recycling (Cain, C. C., Wilson, R. B., and Murphy, R. F. (1991) J. Biol. Chem. 266, 11746-11752). These cells show a rapid loss of cell surface receptors upon temperature shift due to a reduction in the rate of receptor recycling. We show here that, in addition to altered receptor recycling, TfT1.11 cells show three defects in lysosome biogenesis. At the nonpermissive temperature, they 1) redistribute at least one lysosomal enzyme from lysosomes to endosomes, 2) fail to transfer fluid-phase material from early endosomes to later compartments, and 3) fail to accumulate fluid-phase markers due to increased efflux of internalized material. The results suggest that the processes of recycling from the endosome and movement of material from endosomes to lysosomes are tightly linked. PMID- 8244968 TI - Molecular cloning and cDNA sequencing of endoxyloglucan transferase, a novel class of glycosyltransferase that mediates molecular grafting between matrix polysaccharides in plant cell walls. AB - Endoxyloglucan transferase is a novel class of glycosyltransferase recently purified from Vigna angularis (Nishitani, K,, and Tominaga, R. (1992) J. Biol. Chem. 267, 21058-21064). This enzyme is the first transferase identified that catalyzes molecular grafting between polysaccharide cross-links in the cell wall matrix and participates in reconstruction of the network structure in the cell wall. Based on the NH2-terminal amino acid sequence information of the purified transferase, we have here cloned and sequenced cDNAs derived from five different plant species, V. angularis, Triticum aestivum, Arabidopsis thaliana, Lycopersicon esculentum, and Glycine max. In the five plant species, the amino acid sequence of the mature proteins is conserved in the range of 71-90% throughout their length. The consensus sequence for N-linked glycosylation, and four cysteine residues are all conserved in the five species. Thus, the endoxyloglucan transferase protein is ubiquitous among higher plants. The highly conserved DNA sequence will serve as a promising tool for exploring the molecular process by which cell wall construction, and hence cell growth, is regulated. PMID- 8244969 TI - Four contiguous amino acids define the target for streptolydigin resistance in the beta subunit of Escherichia coli RNA polymerase. AB - Streptolydigin (stl), a bacteriostatic inhibitor of transcription elongation, interacts with the beta subunit of Escherichia coli RNA polymerase. We have defined the target for stl resistance using chemical mutagenesis and mutagenic polymerase chain reaction. Mutations resulting in stl resistance are confined to a small cluster of contiguous amino acids, amino acids 543 to 546. These stlr mutants differ from one another in their levels of resistance to stl in vivo and in vitro. We have analyzed two of the mutants, A543V and F545S, for their effects on elongation and termination in vivo and in vitro. Neither affected termination at rho-dependent or rho-independent terminators. These mutants were indistinguishable from wild type in a T7 in vitro elongation assay. F545S, however, did exhibit slower elongation kinetics in a lambda tR1 pausing assay. We conclude that mutations in the stlr region can influence transcription elongation, but that these amino acids are not directly involved in catalysis. PMID- 8244970 TI - Growth factor-inducible alternative splicing of nontransmembrane phosphotyrosine phosphatase PTP-1B pre-mRNA. AB - PTP-1B is a major nontransmembrane phosphotyrosine phosphatase in human cell lines and tissues, but its physiological function(s) and mechanism(s) of regulation are largely unknown. We have found that in human diploid fibroblasts a novel PTP-1B mRNA isoform is produced upon stimulation of quiescent cells with a variety of growth factors. Generation of this isoform requires protein synthesis, suggesting that the product(s) of an immediate early response gene(s) is required. RNase protection and reverse transcription polymerase chain reaction analysis demonstrate that the isoform arises as a consequence of alternative pre mRNA splicing, leading to retention of the last intron. The novel isoform is found on polyribosomes, indicating that it is actively translated, and is variably expressed in human tissues. Sequence analysis indicates that the isoform encodes a PTP-1B protein with an altered C terminus. To our knowledge, this is the first example of growth factor-regulated alternative splicing. PMID- 8244971 TI - Amino acid N-malonyltransferases from mung beans. Action on 1-aminocyclopropane-1 carboxylic acid and D-phenylalanine. AB - 1-Aminocyclopropane-1-carboxylate (ACC) N-malonyltransferase from etiolated mung bean hypocotyls was examined for its relationship to D-phenylalanine N malonyltransferase and other enzymes which transfer malonyl groups from malonyl CoA to D-amino acids. Throughout a 3600-fold purification the ratio of D phenylalanine N-malonyltransferase activity to ACC N-malonyltransferase activity was unchanged. Antibodies raised against purified ACC N-malonyltransferase 55-kDa protein were also able to precipitate all D-phenylalanine-directed activity from partially purified mung bean extracts. The irreversible inhibitors phenylglyoxal and tetranitromethane reduced malonyltransferase activity towards D-phenylalanine to the same extent as that for ACC. In addition, several other D-amino acids, particularly D-tryptophan and D-tyrosine, were able to inhibit action towards both ACC and D-phenylalanine. These lines of evidence suggest that a single enzyme is capable of promoting malonylation of both ACC and D-phenylalanine. Km values for D-phenylalanine and malonyl-CoA were found to be 48 and 43 microM, respectively; these values are 10-fold lower than the corresponding values when ACC was substrate. Coenzyme A was a noncompetitive (mixed type) product inhibitor towards malonyl-CoA at both unsaturated and saturated ACC concentrations. The enzyme was also inhibited uncompetitively at high concentrations of malonyl-CoA. We propose that the enzyme follows an Ordered Bi-Bi reaction pathway, with the amino acid substrate being bound initially. PMID- 8244972 TI - Species and subtype variants of the N-formyl peptide chemotactic receptor reveal multiple important functional domains. AB - The N-formyl peptide receptor (FPR) is a seven transmembrane-domain receptor that mediates trafficking and activation of phagocytic leukocytes in response to N formyl oligopeptides such as fMet-Leu-Phe. cDNAs for high affinity FPRs have been cloned from both human (huFPR) and rabbit (rabFPR). To identify functional domains of FPR, we have studied two structurally related "natural mutants" that are 100-10,000-fold less sensitive than huFPR and rabFPR to fMet-Leu-Phe owing to sequence differences that are located predominantly in the proposed extracellular and transmembrane domains. The first is murine FPR (muFPR, 76% identical to huFPR) whose gene we have now cloned and expressed in Xenopus oocytes; the second is the previously reported human FPR-like 1 receptor (FPRL1R, 69% identical to huFPR) which was used to construct huFPR-FPRL1R chimeras. Comparison of the structure and function of huFPR, FPRL1R, muFPR, rabFPR, and huFPR-FPRL1R chimeras suggests that multiple non-contiguous residues must be apposed by coordinate folding of all of the extracellular and transmembrane domains in order to form the high affinity fMLF-binding site. PMID- 8244973 TI - Characterization of the Streptococcus pneumoniae maltosaccharide regulator MalR, a member of the LacI-GalR family of repressors displaying distinctive genetic features. AB - The gene encoding a transcriptional repressor of the maltosaccharide utilization operons of the Gram-positive bacterium Streptococcus pneumoniae (malR) has been cloned and sequenced. The genetic structure of the locus reveals the presence of an upstream gene necessary for growth on maltotetraose medium (malA). The phenotype of malR- and malA- mutants obtained by interruption of the coding regions suggests that both genes could belong to the same transcription unit. Two copies of a DNA motif consisting of three conserved regions of 59, 42, and 49 nucleotides were found located upstream and downstream of the malA-malR putative operon. These DNA structures are almost identical to the reported Box sequences associated with several genes of S. pneumoniae. The protein encoded by malR was visualized and partially purified after selective expression in Escherichia coli, whereas the product of malA was identified in vitro. The amino acid sequence of MalR displays similarities with the Lac and Gal family of repressors. The highest similarities were found when comparing MalR with the E. coli MalI repressor, which is related with an indirect induction pathway of the maltose regulon. The significance of these similarities is discussed in terms of the possible evolutionary pathways followed by structural and regulatory genes of sugar utilization systems in bacteria. PMID- 8244974 TI - Modifications in a flexible surface loop modulate the isozyme-specific properties of mammalian alkaline phosphatases. AB - We have analyzed to what extent the surface loop domain of alkaline phosphatases (APs) is responsible for isozyme-specific functional properties. Unique AatII and RsrII restriction sites were introduced by site-directed mutagenesis at identical positions in murine tissue-nonspecific AP (TNAP) and human placental AP (PLAP) cDNAs to allow the homologous exchange of the loop domain of the TNAP (T domain) and PLAP (P domain) isozymes and the generation of the reciprocally chimeric molecules PLAP-T and TNAP-P. The introduction of the T loop into PLAP reduced the heat stability of PLAP-T to almost that of TNAP. The domain substitution was accompanied by a conformational change that resulted in the loss of immune reactivity with four of 17 epitope-mapped anti-PLAP monoclonal antibodies. The T and P loops provided stabilization to the side chain of specific uncompetitive AP inhibitors. The introduction of the T domain also conferred collagen-binding properties to PLAP-T accounting for half of the binding affinity of TNAP for collagen, while not affecting PLAP binding to IgG. Our data indicate that the surface loop determines overall enzyme stability, differs conformationally in the various isozymes, and modulates catalytic parameters in the presence of protein ligands, thus, accounting in part for isozyme-specific protein interactions. PMID- 8244975 TI - PAM, a novel plasminogen-binding protein from Streptococcus pyogenes. AB - The ability of group A streptococci to bind human plasminogen and plasmin has attracted interest, because it could provide the bacteria with a mechanism for invasion. M or M-like proteins account for the binding of several plasma proteins to group A streptococci. To investigate whether M or M-like proteins were responsible for the binding of plasminogen to group A streptococci, acid extracted material from a type M53 streptococcal isolate was tested for its ability to bind plasminogen. Indeed, a 42-kDa plasminogen-binding protein was solubilized. Two oligonucleotides homologous with conserved sequences in known M protein genes were used as primers in the polymerase chain reaction, with chromosomal DNA from the M53 isolate. When cloned and expressed in Escherichia coli, a resulting fragment encoded a 43-kDa plasminogen-binding protein. Nucleotide sequence determination of the gene fragment revealed an open reading frame encoding a polypeptide of 43,580 Da, which matched the amino-terminal amino acid sequence of the plasminogen-binding protein extracted from M53 streptococci. The DNA sequence data also proved the relationship of the encoded protein, named PAM, to the M proteins. The plasminogen-binding domain was mapped to the amino terminal third of PAM. Plasminogen absorbed by M53 streptococci or by immobilized PAM could be activated by streptokinase. The results provide further evidence of the diversity of the M protein family and suggest a new mechanism whereby these proteins contribute to the virulence of group A streptococci. PMID- 8244976 TI - pH-sensitive calcium release in triads from frog skeletal muscle. Rapid filtration studies. AB - Triad vesicles from frog skeletal muscle exhibited calcium-induced calcium release highly sensitive to extravesicular pH; calcium induced release at pH > 7.4 but not at pH 6.8. In contrast, triads isolated from rabbit skeletal muscle exhibited significant calcium-induced calcium release at pH 6.8. At pH 7.4, there was no stimulation of calcium release in triads from frog at pCa 7, maximum stimulation at pCa 5, and complete inhibition at pCa 3. Addition of ATP at pCa 5, pH 6.8, induced calcium release with the same high rate constants in both preparations. In triads from frog, ATP-induced calcium release at pCa 5 had the same kinetics at pH 6.8 and 7.4, whereas ATP-induced calcium release at pCa > 8, pH 6.8, was partial, with a decrease in the amounts released but not in rate constants. In contrast, triads from rabbit displayed the opposite behavior, with a decrease in rate constants but not in the amounts of calcium released at pCa > 8, pH 6.8. In triads from frog ATP-induced calcium release decreased at pCa < 4, to reach total inhibition at pCa 2; addition of magnesium (free > 0.5 mM) completely abolished ATP-induced calcium release at pH 6.8, pCa 7 or 5. The potential physiological relevance of these results is discussed. PMID- 8244977 TI - Metabolism of leukotriene B4 by cultured human keratinocytes. Formation of glutathione conjugates and dihydro metabolites. AB - Six previously unidentified leukotriene (LT) B4 metabolites formed during incubation of LTB4 with human keratinocytes in primary culture indicate the importance of the 12-hydroxyeicosanoid dehydrogenase pathway in LTB4 metabolism. The ultraviolet absorption spectra obtained for all keratinocyte metabolites revealed the presence of a conjugated diene structural moiety rather than the conjugated triene structure of LTB4. Metabolites were characterized using fast atom bombardment-mass spectrometry, gas chromatography-mass spectrometry of the pentafluorobenzyl ester, trimethylsilyl ether derivatives and specific degradation reactions. The previously identified 10,11-dihydro-LTB4 and 10,11 dihydro-12-epi-LTB4 were observed as well as 20-OH-10,11-dihydro-LTB4, consistent with the reductase pathway of LTB4 metabolism. This pathway involves initial formation of 12-oxo-LTB4 catalyzed by 12-hydroxyeicosanoid dehydrogenase followed by reduction by delta 10-reductase. The most lipophilic metabolite of LTB4 was identified as 10-hydroxy-4,6,12-octadecatrienoic acid which could result from beta-oxidation reactions of LTB4 following reduction of the 10,11-double bond. One of the most abundant metabolites was characterized as 3,7,14-trihydroxy 8,10,16- docosatrienoic acid which could result from fatty acid elongation following reduction of the 10,11-double bond. Additional abundant LTB4 metabolites were identified that result from glutathione conjugation of 12-oxo LTB4. These were characterized using fast atom bombardment-mass spectrometry and by chemical degradation using hypochlorous acid as well as transpeptidases. These metabolites were identified as 5,12-dihydroxy-6-glutathionyl-7,9,14 eicosatrienoic acid (c-LTB3), 5,12-dihydroxy-6-cysteinyl-glycyl-7,9,14 eicosatrienoic acid (d-LTB3) and 5,12-dihydroxy-6-cysteinyl-7,9,14-eicosatrienoic acid (e-LTB3). We propose that these metabolites result from a 1,8 Michael-type addition of glutathione to the 12-oxo-LTB4 intermediate. PMID- 8244978 TI - In vitro activity of the herpes simplex virus type 1 protease with peptide substrates. AB - Herpes simplex virus type-1 (HSV-1) protease is responsible for proteolytic processing of itself and the virus assembly protein ICP35 (infected cell protein 35). Two proteolytic processing sites within the protease have recently been identified between Ala247 and Ser248 and between Ala610 and Ser611. In this report we demonstrate that peptides corresponding to each of these cleavage sites are substrates for recombinant HSV protease-glutathione S-transferase fusion protein in vitro by high performance liquid chromatography analysis of cleavage reactions. Analysis of the products by fast atom bombardment-mass spectrometry confirmed that cleavage occurred at the expected position between the Ala and Ser residues of the substrate. Peptide cleavage was linear with respect to time and enzyme concentration and proceeded optimally at pH 8.0. A peptide that spans Ala99/Ser100 of the protease but does not correspond to a naturally occurring cleavage site was not a substrate for the protease in vitro confirming that sequence elements outside the conserved dipeptide sequence are required for substrate recognition and cleavage. Identification of P5-P8' as the minimal substrate peptide for the Ala610/Ser611 cleavage site revealed a requirement for residues flanking the conserved core P4-LVNA/S-P1' in substrate recognition and hydrolysis. Kinetic analysis with peptide P5-P8' yielded a Km of 190 microM and kcat of 0.2 min-1. Experiments with a panel of class-specific protease inhibitors were consistent with the protease being a member of the general class of serine proteases. PMID- 8244979 TI - Regulation of protein tyrosine phosphatases by insulin and insulin-like growth factor I. AB - In this study, we have examined the effects of insulin and insulin-like growth factor (IGF)-I on protein tyrosine phosphatase (PTPase) activity in rat L6 skeletal muscle cells. Under basal conditions, about 85% of total cellular PTPase activity was associated with the particulate (Triton X-100-soluble) fraction. Incubation of the cells with 100 nM insulin or IGF-I significantly increased particulate PTPase activity (p < 0.005) without altering activity in the supernatant or Triton X-100-insoluble fractions. Dose responses studies suggested that the effect of each hormone was mediated through its own receptor. PTPase activity was regulated by both acute and chronic insulin and IGF-I treatment. Maximal stimulation by both ligands occurred at 32 h and then declined. By using an antibody and a cDNA specific for PTPase1B, we found that the chronic stimulation of PTPase activity was accompanied by enhanced expression of PTPase1B mRNA and protein. Maximal induction of PTPase1B mRNA and protein by insulin and IGF-I occurred at 12 and 24 h, respectively. Based on these data, it can be suggested that ligand-stimulated PTPase activity might oppose tyrosine kinase mediated insulin or IGF-I signal transmission and thus desensitize cells to long term action by insulin and IGF-I. However, it is also possible that PTPase act as positive mediators of insulin and IGF-I action. PMID- 8244980 TI - Cloning of Chinese hamster DNA topoisomerase I cDNA and identification of a single point mutation responsible for camptothecin resistance. AB - A camptothecin-resistant (DC3F/C-10) Chinese hamster cell line that contains a catalytically altered and camptothecin (CPT)-resistant DNA topoisomerase I (top 1) (Tanizawa, A., and Pommier, Y. (1992) Cancer Res. 52, 1848-1854) and the parent cell line (DC3F) were used to compare top 1 mRNAs and cDNAs. Northern blot analysis showed a single 4.1-kilobase band without quantitative reduction between the two cell lines. We have cloned and sequenced top 1 cDNAs. DC3F and DC3F/C-10 top 1 c-DNA are 3591 and 3626 base pair long, respectively, and encode 767 amino acids. The homology of deduced amino acid sequences between Chinese hamster and mouse or human top 1 are 98.1 and 96.7, respectively. cDNAs from DC3F/C-10 and DC3F cells differ by a single base point mutation (G to A) which results in an amino acid change from Gly505 to Ser (Gly505-->Ser). G505 corresponds to Gly503 of human top 1 cDNA and is located 220 amino acids away from the presumed catalytic Tyr725. The point mutation in the Chinese hamster top 1 is located in a region that is highly conserved among all cloned top 1 cDNAs (plant ATH, vaccinia virus, Shope fibroma virus, Drosophila, Saccharomyces cerevisiae, Schizosaccharomyces pombe, mouse, and Human). A mutation of Asp533 to Gly in this same region has been shown to confer CPT resistance for human top 1. Chinese hamster top 1 protein with a Gly505-->Ser mutation that was expressed in bacteria was resistant to CPT, indicating that this single base mutation is involved in CPT resistance. Our results suggest that the highly conserved region around Gly505 plays an important role in the interactions among top 1, DNA, and CPT. PMID- 8244981 TI - Separate nuclear genes encode cytosolic and mitochondrial nucleoside diphosphate kinase in Dictyostelium discoideum. AB - We have previously isolated cDNA clones for the gip17 gene encoding the cytosolic nucleoside diphosphate (NDP) kinase from Dictyostelium discoideum, and partial cDNAs for guk, a second member of the NDP kinase gene family (Wallet, V., Mutzel, R., Troll, H., Barzu, O., Wurster, B., Veron, M., and Lacombe, M. L. (1990) J. Natl. Cancer Inst. 80, 1199-1202). We now characterize genomic DNA clones for both NDP kinase genes, and we show that guk defines a nuclear-encoded mitochondrial NDP kinase. Isolated D. discoideum mitochondria contain 3% of the total cellular NDP kinase activity. Antibodies which specifically recognize and inhibit the activity of either cytosolic or mitochondrial NDP kinase unambiguously distinguish between these activities. The nascent mitochondrial NDP kinase contains a presequence of 57 amino acids that is removed during import into the organelle as shown by determination of the NH2 terminus of the mature protein from mitochondria. The genes for mitochondrial and cytosolic NDP kinases contain four and two introns, respectively. The positions of the of the introns in the gene for the cytosolic enzyme match exactly the positions of the second and fourth introns in the coding region of its mitochondrial homologue. From these results we conclude that the isozymes diverged from a common ancestor, and we discuss possible phylogenetic pathways for the evolution of cytosolic and organelle NDP kinases. PMID- 8244982 TI - Turnover of the photosystem II D1 protein in higher plants under photoinhibitory and nonphotoinhibitory irradiance. AB - The turnover in vivo of the Photosystem II (PS II) reaction center D1 protein was investigated by [35S] methionine labeling of leaf discs of Brassica napus and subsequent analysis after thylakoid SDS-gel electrophoresis. The rate of D1 protein degradation was found to have a t1/2 of approximately 2 h, at an irradiance corresponding to the growth irradiance. The rate of D1 protein degradation was not increased further by prior photoinhibitory treatment which inactivated 40% of the PS II centers, but the amount of [35S]methionine label incorporated into the D1 protein during 1 h was increased 3-fold. Whether photoinhibited or not, the incorporation of label into the D1 protein increased with irradiance only up to approximately the growth irradiance and above that it decreased with increasing incident light. These data indicate that a high rate of D1 protein degradation occurs not only after photoinhibition but also under conditions where no net decrease in the number of functional PS II centers occurs and that the number of PS II centers that undergo D1 protein turnover varies depending on prior photoinhibitory damage and on incident irradiance. A down regulation of PS II in leaves at high irradiances is discussed. PMID- 8244983 TI - Substitutional and insertional RNA editing of the cytochrome c oxidase subunit 1 mRNA of Physarum polycephalum. AB - The term RNA editing encompasses two types of specific alterations in the coding potential of RNA molecules: base substitution and the insertion (or deletion) of nucleotides. Such changes in RNA sequence can have profound effects on gene expression, and, indeed, most genes in the mitochondria of plants, trypanosomatids, and Physarum appear to require editing for their expression. We describe here the first instance of the utilization of both types of RNA editing in the processing of a single mRNA, that of the mitochondrially encoded cytochrome oxidase subunit I of the acellular slime mold, Physarum polycephalum. Editing of this mRNA includes the insertion of cytidine, guanosine, and uridine residues, as well as the apparent conversion of cytidines to uridines. No edited version of this gene was detected in Physarum DNA, and amino acid alignments suggest that both types of RNA editing are required to produce a functional protein. PMID- 8244984 TI - Low density lipoprotein receptors bind and mediate cellular catabolism of normal very low density lipoproteins in vitro. AB - Very low density lipoproteins (VLDL) are heterogeneous, triglyceride-rich particles that are precursors of low density lipoproteins (LDL). Before conversion to LDL, the majority of VLDL are irreversibly cleared from plasma by uncertain mechanisms. To investigate one potential mechanism for VLDL clearance, we studied the ability of LDL receptors to mediate VLDL uptake in vitro. Small, intermediate, and large VLDL from normolipidemic humans were found to bind and undergo catabolism via LDL receptors on normal human fibroblasts. Binding to cell surfaces was up-regulated by lovastatin, an inducer of LDL receptors. Both LDL and a monoclonal antibody against the LDL receptor (IgG-C7) prevented binding of 125I-VLDL. Also, VLDL binding to mutant fibroblasts lacking LDL receptors was low. Thus, LDL receptors mediated VLDL interactions with cells. Binding affinity decreased near saturation, and the apparent number of high affinity sites decreased with increasing VLDL particle size. Because LDL receptors are small (M(r) 115,000) relative to VLDL (M(r) 9-24 x 10(6)) and are clustered in clathrin coated pits, these findings suggest that steric hindrance becomes an important binding determinant near saturation and are consistent with a lattice model for LDL receptor-ligand interactions. The capacity for cellular catabolism of VLDL decreased with increasing particle size, consistent with a lattice model. The lattice model was also supported by differences between 125I-VLDL binding to cell surfaces and binding to partially purified LDL receptors in solid-phase assays in which steric constraints resulting from clustering in clathrin-coated pits are not present. In both cell-surface and solid-phase assays, VLDL bound via apoE, not apoB-100. Our studies establish that normal VLDL interact with LDL receptors and that steric hindrance due to crowding of particles on clustered LDL receptors is an important determinant of their binding and catabolism. These findings suggest that LDL receptors may participate in normal VLDL clearance in vivo. PMID- 8244985 TI - Bovine lens aldose reductase. pH-dependence of steady-state kinetic parameters and nucleotide binding. AB - The pH-dependence of nucleotide binding and steady-state kinetic parameters of aldehyde reduction and alcohol oxidation catalyzed by bovine lens aldose reductase were studied. The maximal velocity of aldehyde reduction with NADPH and p-chlorobenzaldehyde was pH independent at low pH but decreased at high pH with a pK of 7.6. The V/K of NADPH displayed a bell-shaped dependence on pH and decreased with a pKa of 5.3 and a pKb of 7.5. The dissociation constant of NADPH and 3-acetylpyridine adenine dinucleotide phosphate (3-APADP) increased at low pH with a pK of 5.6-5.8 and at high pH with a pK of 9.4-9.7. The pKi of NADP and NADPH decreased below a pH of 5 and 6.7 and above a pH of 8.5 and 9.7, respectively. The pK of 8.5-9.7 appears to be due to the interaction of the 2' phosphate of the nucleotide with a protonated base, possibly a lysine residue. The maximal velocity of alcohol oxidation was pH independent at high pH but decreased at low pH with a pK of 6.5-7.0, when p-chlorobenzyl alcohol or benzyl alcohol and 3-APADP were used. The amino acid residue for alcohol binding has a pK of 7.5-8.2 and also appears in pKi profiles of sorbinil, a competitive inhibitor versus the alcohol. Large (3-3.5) isotope effects on maximal velocity obtained with benzyl alcohol and 3-APADP suggest that with these substrates the hydride transfer step is rate-limiting and a pK of 6.5-7.0 may be the true pK of the acid-base catalyst, possibly a histidine. PMID- 8244986 TI - Identification of a lysophospholipase C that may be responsible for the biosynthesis of choline plasmalogens by Madin-Darby canine kidney cells. AB - The biosynthesis of choline plasmalogens was investigated in Madin-Darby canine kidney cells to determine the source of the vinyl ether linkage. 1-O-[3H] Alk-1' enyl-2-lyso-sn-glycero-3-phosphoethanolamine was a better precursor than 1-O [3H]alkyl-2-lyso-sn-glycero-3-phosphocholine for the synthesis of 1-O-[3H]alk-1' enyl-2-acyl-sn-glycero-3-phosphocholine; this suggests that the vinyl ether linkage in choline phosphoglycerides originates from ethanolamine plasmalogens. The contribution of N-methylation and base exchange enzymes to choline plasmalogen biosynthesis was assessed using 1-O-[3H]alkenyl-2-lyso-sn-glycero-3 [32P]phosphoethanolamine. While 1-O-[3H]alkenyl-2-acyl-sn-glycero-3 phosphocholine was formed from this precursor, the 32P was lost indicating that N methylation and base exchange enzymes do not contribute significantly to the synthesis of choline plasmalogens. The conversion of a phosphono analog of 1-O [3H]alkyl-2-lyso-sn-glycero-3-phosphoethanolamine, which is resistant to phospholipase D hydrolysis, to 1-O-[3H]alkenyl-2-acyl-sn-glycero-2-phosphocholine was observed demonstrating that phospholipase D is not required for choline plasmalogen biosynthesis. A Mg(2+)-dependent lysophospholipase C activity was detected in microsomes that actively hydrolyzed ether-linked lysophosphoglycerides as well as the lysophosphono analog. To assess the role of lysophospholipase C in shuttling 1-O-alk-1'-enyl-sn-glycerol (alkenylglycerol) from ethanolamine plasmalogens to choline plasmalogens, cells prelabeled with 1-O [3H]alkenyl-2-lyso-sn-glycero-3-phosphoethanolamine were treated with 12-O tetradecanoylphorbol-13-acetate. This resulted in the rapid deacylation of 1-O [3H]alkenyl-2-acyl-sn-glycero-3-phosphoethanolamine to 1-O-[3H]alkenyl-2-lyso-sn glycero-3-phosphoethanolamine and the subsequent generation of 1-O [3H]alkenylglycerol. A concomitant 2-3-fold increase in 1-O-[3H]alkenyl-2-acyl-sn glycero-3-phosphocholine was observed. These studies suggest that the alkenyl linkage in choline phosphoglycerides may originate from 1-O-alkenyl-2-lyso-sn glycero-3-phosphoethanolamine through an enzymatic pathway involving lysophospholipase C to generate alkenylglycerol that is subsequently converted to choline plasmalogens. PMID- 8244987 TI - Appendix. Analysis of tricarboxylic acid cycle using mass isotopomer ratios. PMID- 8244988 TI - Kinetics and regulation of three cloned mammalian Na+/H+ exchangers stably expressed in a fibroblast cell line. AB - The kinetics and second messenger regulation of three cloned mammalian intestinal Na+/H+ exchangers were studied using fluorometric techniques. These exchangers, NHE1, NHE2, and NHE3, were stably expressed in PS120 fibroblasts, which lack an endogenous Na+/H+ exchanger. H+ kinetic data indicated cooperativity by internal protons, with Hill coefficients of approximately 2 for all three isoforms. In contrast, Na+ kinetic data fit Michaelis-Menten kinetics, with Km (Na+) 15-18 mM and a Hill coefficient of approximately 1. The exchangers were all activated by growth factors and thrombin; in NHE1 these agonists increased the apparent affinity for intracellular H+, but did not change Vmax, while for NHE2 and NHE3 the effect was on Vmax alone. Phorbol ester stimulated NHE1 and NHE2, but inhibited NHE3 with a decrease in Vmax. ATP-depletion decreased Vmax and the apparent affinity for H+ for all three isoforms, and reduced the Hill coefficient to approximately 1, suggesting that a basal level of phosphorylation was required for the cooperativity. The differences in kinetics and second messenger regulation suggest that the NHE isoforms may serve different cellular functions. The up- and down-regulation of NHE3 by kinases indicates that this isoform may be involved in a specialized function such as Na+ absorption. PMID- 8244989 TI - Functional properties of the rat Na/H exchanger NHE-2 isoform expressed in Na/H exchanger-deficient Chinese hamster ovary cells. AB - The primary structure and functional expression of the rat Na/H exchanger (NHE) NHE-2 isoform has recently been reported (Wang, Z., Orlowski, J., and Shull, G. E. (1993) J. Biol. Chem. 268, 11925-11928). To further characterize some of its functional properties, biochemical and pharmacological analyses were performed on exchanger-deficient Chinese hamster ovary cells (AP-1) that had been stably transfected with a full-length NHE-2 cDNA. Transport activity for NHE-2 was assayed by measuring amiloride-inhibitable 22Na+ influx following an acute intracellular acid load. Pharmacological analyses revealed that NHE-2 had a relatively high affinity for amiloride and some of its analogues. The most potent analogue was 5-(N-ethyl-N-isopropyl)amiloride (EIPA) (K0.5 = 79 nM), followed by 5-(N,N-dimethyl)amiloride (DMA) (K0.5 = 250 nM), amiloride (K0.5 = 1.4 microM), and benzamil (K0.5 = 320 microM). Nonamiloride compounds known to inhibit the activity of other Na/H exchanger isoforms also inhibited NHE-2 with the following order of potency: clonidine (K0.5 = 42 microM) > harmaline (K0.5 = 330 microM) approximately cimetidine (K0.5 = 330 microM). Biochemical analyses showed that the extracellular Na+ dependence of NHE-2 followed simple, saturating Michaelis Menten kinetics with an apparent affinity constant for Na+ (KNa) of 50 mM. In contrast, intracellular H+ appeared to activate NHE-2 by a positive cooperative mechanism with an apparent half-maximal activation value of pK 6.90. Other cations, such as extracellular Li+ and H+, acted as competitive inhibitors of 22Na+ influx by NHE-2, with apparent Ki values of 3.0 mM and 10 nM, respectively. In contrast, extracellular K+ had no effect on the transport activity of NHE-2. These results indicated that the rat NHE-2 cDNA encodes a functional Na/H exchanger isoform with distinct properties compared to rat NHE-1 and -3. PMID- 8244990 TI - Comparison of kinetic properties between two mammalian ras p21 GDP/GTP exchange proteins, ras guanine nucleotide-releasing factor and smg GDP dissociation stimulation. AB - The mammalian counterpart of the yeast ras p21 GDP/GTP exchange protein CDC25, ras GRF, was expressed in Escherichia coli and purified, and its kinetic properties were compared with those of another mammalian ras p21 GDP/GTP exchange protein, smg GDS. ras GRF was active on Ki- and Ha-ras p21s but inactive on rap1A p21, rhoA p21, rac1 p21, and rab3A p25, whereas smg GDS was active on Ki-ras p21, rap1A p21, rhoA p21, and rac1 p21 but inactive on Ha-ras p21 and rab3A p25. The Kcat values of ras GRF and smg GDS for Ki-ras p21 as a common substrate were calculated to be 1.2 and 0.37 nmol/min/nmol, respectively. The Km values of ras GRF and smg GDS for Ki-ras p21 were 680 and 220 nM, respectively. rasGRF was slightly active on post-translationally unprocessed Ki-ras p21 but much more effective on post-translationally processed Ki-ras p21 than on post translationally unprocessed Ki-ras p21. smg GDS was active on post translationally processed Ki-ras p21 but inactive on post-translationally unprocessed Ki-ras p21. Moreover, as described for smg GDS, ras GRF showed a potency to inhibit the binding of Ki-ras p21 to membrane and to induce the dissociation of prebound Ki-ras p21 from the membrane. These results indicate that ras GRF and smg GDS show apparently similar kinetic properties except for the different substrate specificities and the requirement of the post translational processing of Ki-ras p21. PMID- 8244991 TI - The inactivation of dihydroxy-acid dehydratase in Escherichia coli treated with hyperbaric oxygen occurs because of the destruction of its Fe-S cluster, but the enzyme remains in the cell in a form that can be reactivated. AB - The enzyme dihydroxy-acid dehydratase previously has been shown to be inactivated in vivo in Escherichia coli within minutes of exposure to hyperbaric O2. In this paper, we show its inactivation is due to the destruction of its catalytically active [4Fe-4S] cluster. The inactivation is not followed by an appreciable decrease in the amount of dihydroxy-acid dehydratase protein as determined by Western blots. Thus, the protein from the inactivated enzyme remains unproteolyzed in the cells. Dihydroxy-acid dehydratase activity recovers after the cells treated with hyperbaric O2 are returned to ambient oxygen. Since this recovery in activity is not accompanied by a significant increase in dihydroxy acid dehydratase protein and is not prevented by chloramphenicol, it appears primarily to be due to reactivation of the previously inactivated enzyme. The reactivation occurs by reconstitution of the enzyme's Fe-S cluster. These results demonstrate that this enzyme can cycle between forms in which the Fe-S cluster is either present or absent. The facile ability to cycle between these two forms would be compatible with a regulatory role in addition to a catalytic role for this enzyme. PMID- 8244992 TI - Tendon hypertrophy is associated with increased hydroxylation of nonhelical lysine residues at two specific cross-linking sites in type I collagen. AB - This study was designed to investigate whether the changes in lysine hydroxylation known to occur in hypertrophic tendon occur randomly or at specific lysine residues in the type I collagen molecule. Peptides corresponding to the two known major cross-linking sites of type I collagen (a lysine (or hydroxylysine) at position 9N cross-linked to a hydroxylysine at 930 and a lysine (or hydroxylysine) at position 16C cross-linked to a hydroxylysine at position 87) were prepared by collagenase digestion, size fractionation, and separation by high performance liquid chromatography from normal chicken tendon and from chicken tendon subjected to increased tensile load as a result of muscle hypertrophy. The ratio of the difunctional cross-links dihydroxylysinonorleucine to hydroxylysinonorleucine in normal tendon is 0.75:1; this ratio is increased to 6:1 in hypertrophic tendon. The dihydroxylysinonorleucine to hydroxylysinonorleucine ratio is increased to the same extent in samples of the purified cross-linked peptides derived from both the N-terminal and C-terminal lysine aldehyde residues. On the other hand, the relative hydroxylysine content of preparations of the pooled larger helical peptides obtained by cyanogen bromide digestion of normal and hypertrophic tendons was essentially identical. These results demonstrate that there is a specific increase in hydroxylation of only the N- and C-terminal non-helical lysine residues that participate in the formation of the reducible difunctional cross-links of type I collagen in hypertrophic tendon, while the extent of hydroxylation of lysine residues in the helical regions is not affected. The specific mechanism by which the enzyme lysyl hydroxylase acting on its substrate can distinguish between lysine residues destined to be in non-helical versus helical regions in a nascent collagenous peptide that has not yet attained its final secondary structure remains to be defined. PMID- 8244993 TI - Drosophila proteasome Dm25 subunit substitutes the mouse MC3 subunit in hybrid proteasomes. The N-terminal domain is essential for subunit incorporation. AB - The proteasome is a multisubunit 20 S proteinase complex involved in ubiquitin dependent and -independent intracellular protein metabolism. Individual subunits of the alpha- and beta-type share extensive sequence homology and are encoded as members of two related and evolutionarily conserved gene families. Due to the lack of viable deletion mutants of essential alpha-type proteasome subunits in higher eukaryotes, an identification and analysis of potentially homologous subunits of different species was so far not possible. It is shown here that the novel Drosophila alpha-type Dm25 subunit can be incorporated into mouse proteasomes of stably transfected NIH 3T3 cells. The Dm25 subunit is able to substitute the mouse MC3 alpha-type subunit in proteasomes, indicating a high structural and possibly also functional homology of the two subunits. In contrast and pointing at the importance of the slightly hydrophobic N-terminal region stabile expression of a Dm25 subunit, which is truncated at its N terminus and lacks PROS box I, results in a subunit which cannot be incorporated into mouse proteasomes. The ability to form hybrid proteasomes involving essential nondeletable subunits now opens the possibility for structural and also functional analysis of such subunits by mutagenesis in higher eukaryotes. PMID- 8244994 TI - Regulation of interleukin 8 gene expression by oxidant stress. AB - Interleukin 8 (IL-8) is a recently described cytokine that functions as a potent neutrophil chemoattractant and activator. We sought to examine the link between the generation of reactive oxygen intermediates (ROI) and the regulation of IL-8 gene expression to specifically test the hypothesis that ROI would induce production of IL-8 mRNA and protein. In lipopolysaccharide-stimulated human whole blood, the OH radical scavenger dimethyl sulfoxide (Me2SO) dramatically inhibited (approximately 90%) IL-8 production, but had minimal effects on the production of tumor necrosis factor, interleukin 1 beta (IL-1), and IL-6. To determine whether NADPH-oxidase-generated free radicals were critical in the regulation of IL-8, studies were performed using blood from patients with chronic granulomatous disease. In both normal individuals and patients with chronic granulomatous disease, production of IL-8 could be initiated with lipopolysaccharide, phytohemagglutinin, or aggregated immune complexes, and this production could be inhibited by Me2SO (1% v/v). To examine if oxidant stress represents a ubiquitous mechanism for the induction of IL-8, experiments were performed in cultured cell lines. In the human hepatoma cell line Hep-G2, Me2SO dose-dependently inhibited tumor necrosis factor-stimulated IL-8 production, with a 74 +/- 1% reduction observed at a Me2SO concentration of 1%. Direct exposure to ROI demonstrated that H2O2 stimulated IL-8 production in a dose-dependent manner in Hep-G2 cells, A549 pulmonary type II epithelial cells, and human skin fibroblasts; this induction could be prevented by addition of catalase. The production of IL-8 appeared to be specific to an oxidant stress since exposure of the cells to heat shock or chemical stress did not induce expression of IL-8. These studies demonstrate that oxidant stress is an important regulator of IL-8 gene expression and support the hypothesis that low levels of ROI may serve to initiate IL-8 production which then serves to recruit neutrophils to sites of inflammation. PMID- 8244995 TI - Molecular characterization of a multi-promoter gene encoding a chicken filamin protein. AB - We report the cloning and sequencing of cDNA encoding a chicken filamin protein. The 2,567 amino acid protein contains an NH2-terminal 267 amino acid actin binding domain followed by a series of 24 repeating units that are each approximately 95 amino acids in length. The overall primary structure of filamin closely resembles that of human actin-binding protein (ABP). However, filamin lacks a 24-amino-acid insertion prior to repeat 16 that is contained within ABP. This region of human ABP is a site of calpain cleavage and is thought to confer flexibility on the molecule. Hence, it is possible that the properties of actin gels formed with either human ABP or filamin reflect the presence or absence of this insertion. Filamin is encoded within a multi-promoter transcription unit. A downstream filamin promoter (Fil1) resembles those of certain housekeeping genes and has a putative binding site for the transcription factor E2F. Thus, transcription from this promoter may be linked to the cell cycle. A second filamin promoter (Fil2) is located at least 8 kilobases upstream from the Fil1 promoter. This structural arrangement suggests that regulation of filamin gene expression is likely to be complex. PMID- 8244996 TI - RNA polymerase II transcription factor SIII. I. Identification, purification, and properties. AB - A transcription factor that stimulates synthesis of accurately initiated transcripts by RNA polymerase II has been identified and purified to apparent homogeneity from rat liver nuclear extracts. This factor, which we designate SIII, has a native molecular mass of approximately 140 kDa and is composed of three polypeptides of 110, 18, and 15 kDa. All three polypeptides are required to reconstitute SIII transcriptional activity. SIII appears distinct from previously identified mammalian transcription factors. PMID- 8244997 TI - The role of NF-kappa B and NF-IL6 transactivating factors in the synergistic activation of human serum amyloid A gene expression by interleukin-1 and interleukin-6. AB - To understand the mechanisms by which large increases in serum amyloid A (SAA) occur during the acute phase response, human hepatoma cells were transfected with SAA2 gene reporter plasmids and stimulated with combinations of cytokines. Although interleukin-1 (IL-1) and interleukin-6 (IL-6) stimulated transcription from this promoter individually, addition of both mediators produced a response between two and nine times greater than the expected additive response. This synergistic activation was dependent on the integrity of at least two cis-acting sequences in the SAA2 enhancer. The SAA2 NF-kappa B site was required functionally for the response to both IL-1 and IL-6 alone as well as for synergistic activation; however, IL-6 did not directly induce binding of nuclear proteins to the NF-kappa B sequence. A NF-IL6 site was required for full induction by IL-1 and IL-6, and also mediated strong transactivation by recombinant NF-IL6. Furthermore, transfected NF-IL6 synergized strongly with co transfected NF-kappa B, particularly with RelA (p65). However synergy between IL 1 and IL-6 was only partly reduced by mutation of the NF-IL6 site, indicating further levels of interaction in addition to the NF-kappa B/NF-IL6 cooperativity. PMID- 8244998 TI - Random mutagenesis of the sheep Na,K-ATPase alpha-1 subunit generates a novel T797N mutation that results in a ouabain-resistant enzyme. AB - Site-directed mutagenesis of the Na,K-ATPase has provided a rational approach to identify several amino acids that appear to be involved in ouabain sensitivity. In order to identify additional amino acids that play a role in ouabain binding, we used formic acid to randomly mutagenize a cDNA cassette encoding amino acids 691-946 of the sheep Na,K-ATPase alpha subunit. This was then used to replace the wild type cDNA cassette in the full-length cDNA, and pools of mutants were electroporated into HeLa cells. Ouabain-resistant cells were selected in 0.5 microM ouabain, and the polymerase chain reaction was used to amplify the region corresponding to the mutagenized cassette from the genomic DNA of the resistant cells. Sequence analysis of the polymerase chain reaction product revealed a single amino acid substitution, T797N. Site-directed mutagenesis was used to reproduce this change in the wild type sheep alpha subunit, and this construct was able to confer resistance to HeLa cells, verifying the role of the mutation in ouabain resistance. The mutant sheep enzyme was found to be as resistant to ouabain as is the rat Na,K-ATPase. These data suggest that T797N, predicted to be in the fifth putative transmembrane domain, is involved in the interaction between ouabain and the Na,K-ATPase. PMID- 8244999 TI - Glucuronidation of hyodeoxycholic acid in human liver. Evidence for a selective role of UDP-glucuronosyltransferase 2B4. AB - Monospecific polyclonal antibodies were raised against a variable amino-terminal domain (amino acids 14-150) of a human liver form of UDP-glucuronosyl-transferase conjugating bile acids, UGT2B4 (Jackson, M. R., McCarthy, L. R., Harding, D., Wilson, S., Coughtrie, M. W., and Burchell, B. (1987) Biochem. J. 242, 581-588), expressed as a fusion protein in Escherichia coli. The antibodies were able to recognize the protein, stably expressed in a genetically engineered eukaryotic V79 cell line, against which they were directed. The specificity of these antibodies allowed their use for analyzing the substrate specificity of this isoform in human liver, as well as for determining its contribution to the total hepatic and extra-hepatic glucuronidation of hyodeoxycholic acid. Western blot analysis of microsomal proteins demonstrated the presence of UGT2B4 exclusively in human liver and not in human kidney. In human liver microsomes, the antibodies were able to inhibit and precipitate up to 90% of the total hyodeoxycholic acid 6 O-glucuronidation activity, but had no effect on activities toward several other substrates, such as phenols, bilirubin, or other bile acids, especially hyocholic acid and the steroids 4-hydroxyesterone and estriol. Moreover, Western blot analysis and immunoinhibition studies of human liver microsomes from healthy patients and from patients presenting liver diseases revealed a good correlation between the glucuronidation rate of hyodeoxycholic acid and the UGT2B4 expression level. The absence of immunoinhibition of hyodeoxycholic acid conjugation with UDP sugars other than UDP-glucuronic acid suggests the involvement of different enzymatic systems in the glucosidation and xylosylation of hyodeoxycholic acid. Altogether, the results provided strong evidence for the specific and predominant involvement of UGT2B4 in the 6-O-glucuronidation of this bile acid via a UDP glucuronic acid-dependent mechanism. PMID- 8245000 TI - Differential regulation of inducible and constitutive prostaglandin endoperoxide synthase in osteoblastic MC3T3-E1 cells. AB - Regulation of mRNA levels for the constitutive and inducible prostaglandin endoperoxide synthases, PGHS-1 and PGHS-2, was examined in murine osteoblastic MC3T3-E1 cells. Serum induction of PGHS-2 mRNA levels was rapid, transient, increased by cycloheximide, and inhibited 72% by cortisol. The cortisol inhibition was blocked by cycloheximide. Serum stimulation of PGHS-1 mRNA was slower, decreased by cycloheximide, and inhibited 28% by cortisol. Increased prostaglandin E2 (PGE2) production and induction of PGHS-2 immunoreactive protein paralleled changes in PGHS-2 mRNA. PGHS-2 mRNA was induced at 2 h in serum-free cells by transforming growth factor-beta (TGF-beta), phorbol 12-myristate 13 acetate, and, to a lesser extent, by forskolin. The combination of phorbol 12 myristate 13-acetate and forskolin was synergistic. TGF-beta induction was prolonged compared with serum, inhibited 67% by cortisol, and the inhibition was not blocked by cycloheximide. TGF-alpha had little effect on PGHS-2 mRNA at 2 h, but the combination of TGF-beta and TGF-alpha was synergistic for PGHS-1 and PGHS 2. PGE2 itself induced PGHS-2 mRNA, and inhibition of PGE2 production decreased the serum induction by 55%, suggesting an important role for autoamplification. The rapidity and amplitude of changes in PGHS-2 suggest that it may be involved in bone responses to acute stresses, such as mechanical strain, inflammation, and injury. PMID- 8245001 TI - Demonstration that a kifunensine-resistant alpha-mannosidase with a unique processing action on N-linked oligosaccharides occurs in rat liver endoplasmic reticulum and various cultured cells. AB - A novel alpha-mannosidase has been identified in rat liver endoplasmic reticulum (ER) which at neutral pH processes the Man9GlcNAc oligosaccharide of glycoproteins by specifically cleaving the terminal mannose residue of the alpha 1,6-linked chain to yield Man8GlcNAc, isomer C. This enzyme accounted for about half of the total ER alpha-mannosidase activity and was fully active at the concentration (0.25 microM) of kifunensine (KIF) completely inhibitory to the action of the ER enzyme which by removing the terminal sugar of the middle chain converts Man9GlcNAc to Man8GlcNAc isomer B; both ER enzymes, however, were inhibited in a similar manner by 1-deoxymannojirimycin (IC50 = 0.2 mM) and their action could not be distinguished with this agent. The KIF-resistant mannosidase which functioned optimally in the presence of 0.1-0.5% Triton X-100 did not show the high susceptibility to EDTA demonstrated by the KIF-sensitive enzyme and unlike the latter had the capacity to hydrolyze p-nitrophenyl-alpha-D-mannoside (Km = 0.45 mM); it had no specific cation requirements, but its activity was greatly reduced in the presence of Zn2+. In isolated ER membranes as well as in intact carbonyl cyanide m-chlorophenylhydrazone-treated cells, the processing pattern was substantially different in the presence of KIF than in its absence; while in the latter instance Man9GlcNAc was readily converted to Man6GlcNAc, the KIF-resistant enzyme was limited in its capacity to go beyond Man8GlcNAc. The KIF resistant alpha-mannosidase was found in substantial amounts in all cell lines examined (HL-60, BW5147.3, MOLT-4, K-562, HepG2, Chinese hamster ovary, F-9, Madin-Darby canine kidney, FRTL-5). The finding that mannose removal from N linked oligosaccharides can be initiated in two distinctive manners substantially broadens our concept of the processing events which can occur before a glycoprotein reaches the Golgi complex or to which ER resident molecules can be exposed. PMID- 8245002 TI - Membrane assembly of the outer membrane protein OmpA of Escherichia coli. AB - The membrane part (residues 1 to approximately 170) of the 325-residue Escherichia coli outer membrane protein OmpA is thought to exist in the membrane as an 8-stranded beta-barrel, subdividing this part into four segments. The influence of proline residues on membrane assembly of the protein has been studied. These were introduced, using site-directed mutagenesis, into each of seven of the antiparallel beta-strands. One important parameter for allowing or not allowing membrane assembly was the potential H beta (i) which is the potential to form an amphiphilic beta-strand. When H beta (i) remained unaltered, 2 prolines were tolerated. Lowering H beta (i) in most cases caused failure of assembly when 2 such residues were present. An insert of 10 residues, including 3 prolines, did not alter H beta(i) and was tolerated, but caused "looping out" of the strand to the outer face of the membrane; displacement to its inner side would not have allowed for an amphiphilic beta-strand. Thus, a beta-structured protein is as adaptable as it has been shown for an alpha-helix. The wild type segment order 1-2-3-4 has been changed to 1-3-3-4 and 1-4-3-4. Since the proteins were found associated with the outer membrane but could not be incorporated into it, it appears that sorting is less sensitive to alterations than assembly. A regulatory circuit was affected (missense mutants of outer membrane proteins can cause inhibition of synthesis of other such proteins); expression of the two rearranged genes effected a strong inhibition of synthesis of the unrelated porins OmpC and F as well as that of the maltoporin LamB and wild type OmpA. Hence, outer membrane proteins are designed not only for efficient membrane assembly but also for proper regulation of their synthesis. PMID- 8245003 TI - Multiple signal transduction pathways induce phosphorylation of serines 16, 25, and 38 of oncoprotein 18 in T lymphocytes. AB - A multitude of external signals induce extensive phosphorylation of Oncoprotein 18 (Op18), which suggests a putative role for this protein in signal transduction. We have recently identified two distinct proline-directed kinase families that phosphorylates Op18 with overlapping but distinct site preference. These two kinase families, mitogen-activated protein (MAP) kinases and cyclin dependent cdc2 kinases, are involved in receptor- and cell cycle-regulated phosphorylation events, respectively. In the present study, site-specific phosphorylation of Op18 in response to stimulation of the antigen receptor associated CD3 complex was analyzed in the Jurkat T cell-line. The results show that CD3-induced phosphorylation of Ser-25 of Op18, which is the primary MAP kinase phosphorylation site, can be induced by an apparently protein kinase C (PKC)-independent signal transduction pathway. We also demonstrate that Ser-16 of Op18 is specifically phosphorylated in response to the Ca2+ signal generated by CD3 stimulation or by the Ca2+ ionophore ionomycin. Ser-16 phosphorylation occurs independently of both PKC and MAP kinase activation. Using site-specific Op18 mutants and tryptic phosphopeptide mapping, we show that phosphorylation of Ser 16 of Op18 together with Ser-25, or Ser-25 and Ser-38, generates two Op18 phosphoisomers showing a dramatic electrophoretic retardation. In conclusion, site-mapping studies of Op18 reveal that CD3 stimulation results in an apparently PKC-independent activation of both the MAP kinase and a Ca(2+)-regulated kinase pathway, which results in phosphorylation of distinct sites of Op18. The data also pinpoints the specific phosphorylation events that result in electrophoretic retardation of Op18. PMID- 8245004 TI - A rat homolog of the Drosophila enhancer of split (groucho) locus lacking WD-40 repeats. AB - In Drosophila, neurogenic loci function in defining cellular fate by interpreting the identity of other cells in the immediate environment. To begin studies of mammalian homologs of these genes, we have isolated two rat homologs of the neurogenic locus Enhancer of split. The protein encoded by the Drosophila Enhancer of split locus is complex and contains five distinct regions based on amino acid composition. One region contains six WD-40 repeats, which were first described in the beta subunit of the heterotrimeric guanine nucleotide-binding protein. One of the rat cDNAs we isolated, R-esp1, encodes a novel form that lacks the WD-40 repeating units. Data are presented demonstrating that the R-esp1 cDNA is a full-length clone encoding an expressed 24-kDa protein. Antibodies raised against this protein stain the nucleus of both PC-12 and GH3 cells. The second clone, R-esp2, encodes a full-length homolog containing WD-40 repeats. The hydrodynamic properties of in vitro translated R-esp1 and R-esp2 proteins indicate that they do not stably self-associate or form heterodimers. A model is presented for the possible role of the R-esp1 protein in the negative regulation of Enhancer of split proteins containing WD-40 repeats. PMID- 8245005 TI - Probing the active site of human aldose reductase. Site-directed mutagenesis of Asp-43, Tyr-48, Lys-77, and His-110. AB - Structural models of human aldose reductase complexed with NADPH have revealed the apposition of C4 of the nicotinamide ring with tyrosine 48 and histidine 110, suggesting that either of these residues could function as the proton donor in the reaction mechanism. Tyrosine 48 is also part of a hydrogen-bonding network that includes lysine 77 and aspartate 43. In order to study the potential catalytic roles of these 4 residues, we evaluated the kinetic properties of mutants containing structurally conservative replacements at these sites. Enzymatic activity was undetectable when Tyr-48 was mutated to phenylalanine (Y48F) although affinity for NADPH was unchanged. In contrast, a mutant containing asparagine substituted for His-110 (H110N) was characterized by an almost 80,000-fold increase in Km, but only about a 14-fold reduction in kcat measured with D-glyceraldehyde. Modest changes in catalytic properties were observed in the mutant containing aspartate 43 substituted with asparagine (D43N): Km for aldehyde substrates was elevated up to 17-fold, and kcat decreased less than 16-fold. However, the Kd(NADP) values for D43N were about 5 times higher than those for wild type. Mutant enzyme containing methionine substituted for lysine 77 (K77M) was up to 1,460-fold less active than the wild type. These results are consistent with Tyr-48 acting as the acid-base catalyst in human aldose reductase and confirm the importance of Asp-43, Lys-77, and His-110 to the structure and function of the active site. PMID- 8245006 TI - The transactivation potential of variant hepatocyte nuclear factor 1 is modified by alternative splicing. AB - Two forms of the transcription factor vHNF1 (HNF1 beta or LFB3) have been previously described, derived by alternative splicing from a common premessenger RNA, and have been called vHNF1-A and vHNF1-B. vHNF1 proteins share a homologous homeo-related DNA-binding domain with the HNF1 protein, initially characterized as a liver-restricted transcription factor, and bind to a similar sequence motif. Here we demonstrate that vHNF1-A is a stronger transactivator than vHNF1-B when assayed in transient transfections using two different promoters. vHNF1-A also binds DNA with a higher affinity suggesting that a region of the protein located immediately upstream of the homeodomain can modulate the protein/DNA interaction and transactivation. Both vHNF1 transcripts were found at a constant ratio in every tissue where vHNF1 expression could be detected, using a quantitative reverse transcriptase-polymerase chain reaction. PMID- 8245007 TI - In vivo phosphorylation sites in fetal and adult rat tau. AB - Fetal tau and tau in paired helical filaments show similar immunoreactivities with several phosphorylation-dependent paired helical filament-polyclonals and monoclonals, suggesting that the two molecules share several distinct phosphorylated epitopes. To make clear the similarities and differences between the two, we have undertaken work to identify the in vivo phosphorylation sites in fetal rat tau. We have approached this problem by identifying phosphopeptides by means of mass spectrometry and sequencing of those phosphopeptides after modification with ethanethiol. Although remarkable heterogeneity was present, fetal tau was found to bear at most 10 phosphates at Ser-189, Ser-190, Ser-193, Ser-226, Ser-387, Ser-395, Thr-172, Thr-222, and, presumably, Ser-391 and Thr-208 (numbering is according to the longest form of rat tau; Kosik, K. S., Orecchio, L. D., Bakalis, S., and Neve, R. L. (1989) Neuron 2, 1389-1397). In contrast, adult rat tau was much less phosphorylated; only Thr-172, Ser-190, Ser-193, Thr 222, and Ser-395 were phosphorylated to a slight-to-moderate extent. All these sites except for Ser-189 and Ser-391 were followed by Pro residues. Thus, tau is an in vivo substrate for proline-directed protein kinase(s), and its phosphorylation state is developmentally regulated. PMID- 8245008 TI - cDNA cloning, expression during development, and genome mapping of PSY2, a second tomato gene encoding phytoene synthase. AB - The enzyme phytoene synthase (Psy) catalyzes the formation of phytoene, an intermediate in the carotenoid biosynthesis pathway. Expression of a previously described gene (PSY1) is induced by fruit ripening in tomato (Lycopersicon esculentum). We describe here the cloning of a partial cDNA for PSY2, a gene related to PSY1. A plasmid containing the PSY2 coding region under control of a bacterial promoter complements a Rhodobacter capsulatus phytoene synthase mutant, indicating that this gene has the capacity to encode an active enzyme. We used reverse transcriptase-polymerase chain reaction followed by digestion with a restriction enzyme to determine that both PSY1 and PSY2 are expressed during tomato development. PSY1 transcripts predominate in seedlings and in late stages of fruit ripening, whereas PSY2 transcripts are relatively more abundant in mature leaves. Both genes are expressed under photooxidative conditions induced by treatment with the carotenoid biosynthesis inhibitor Norflurazon. We used polymerase chain reaction-based restriction fragment length polymorphisms and alien addition lines to map PSY2 to chromosome 2. We conclude that PSY2 is a second tomato gene encoding phytoene synthase. PMID- 8245009 TI - Binding affinity and inhibitory properties of a single-chain anti-T cell receptor antibody. AB - The antigen-specific function of T cells is mediated through a surface receptor (alpha beta). Antibodies to the receptor can either inhibit or activate the T cell depending on the multimeric form of the antibody and the isotype of the antibody constant regions. To further examine and eventually control these antagonistic properties, an anti-receptor antibody (1B2) was engineered by linking the variable region genes of the heavy and light chains to form a single chain binding domain (scFv). The native 1B2 antibody recognizes determinants on the variable regions of both the alpha and beta subunits of the T cell receptor. The scFv gene was expressed in Escherichia coli, and after folding by dilution of solubilized inclusion bodies, scFv protein was purified as both a 29,000-Da monomer and a noncovalently associated 54,000-Da dimer. The binding affinity of the monomer (KD = 17 nM) was 20-fold lower than that of 1B2 Fab fragments (KD = 0.9 nM). In contrast, the binding affinity of the dimer (KD = 1.6 nM) was approximately 2-fold lower than that of 1B2 Fab fragments. Both forms were able to inhibit recognition by the T cell, but the dimer was significantly more effective than the monomer. Inhibition was observed for T cell recognition of either the conventional ligand (a peptide bound to a class I product of the major histocompatibility complex) or the "superantigen" ligand (staphylococcal enterotoxin B bound to a class II product of the major histocompatibility complex). The results suggest that scFv dimers are the most active form of at least some scFv preparations and that the dimers may have the most potential for inhibiting detrimental in vivo T cell activity. PMID- 8245010 TI - Human rhinovirus-14 protease 3C (3Cpro) binds specifically to the 5'-noncoding region of the viral RNA. Evidence that 3Cpro has different domains for the RNA binding and proteolytic activities. AB - Protease 3C (3Cpro) encoded by human rhinovirus type 14 was purified from recombinant Escherichia coli and shown to bind specifically to the 5'-terminal 126 nucleotides of the viral RNA (126 RNA) in addition to efficiently cleaving a synthetic peptide in trans. The binding of 3Cpro to the viral RNA may be required for the initiation of plus strand viral RNA synthesis, suggesting a second non proteolytic function for 3Cpro. Single amino acid substitutions were generated in 3Cpro at residues that are highly conserved among picornaviruses or that lie within the putative catalytic triad. Conservative changes at Asp-85 (D85E and D85N) destroyed the ability of 3Cpro to bind specifically to the 126 RNA, whereas the D85N mutation resulted in almost wild-type levels of proteolytic activity. Conversely, substitutions at His-40, Glu-71, or Cys-146 (H40D, E71A, or C146S) gave proteolytically inactive mutants that bound to the 126 RNA. These results suggest that the highly conserved Asp-85 is essential for specific binding to the 126 RNA, but is unlikely to function in proteolysis as the acidic member of the catalytic triad. Moreover, 3Cpro appears to have different domains for the RNA binding and proteolytic activities. PMID- 8245011 TI - Bile salt-dependent lipase biosynthesis in rat pancreatic AR 4-2 J cells. Essential requirement of N-linked oligosaccharide for secretion and expression of a fully active enzyme. AB - This study used the rat pancreatic AR 4-2 J cell line as a model system to investigate the role of glycosylation in bile salt-dependent lipase (BSDL) secretion and esterolytic activity. Results indicated that AR 4-2 J cells synthesized BSDL, the esterolytic activity of which was the greater part of the cell activity toward 4-nitrophenylcaproate. The protein thus expressed was glycosylated and had a molecular mass approximately 74 kDa. Exposure of these cells to tunicamycin significantly decreased [3H]mannose incorporation, while [35S]methionine incorporation in trichloroacetic acid-precipitable material was not modified. Tunicamycin treatment of AR 4-2 J cells lead to a lower molecular mass form (approximately 70 kDa) of BSDL which did not incorporate [3H]mannose. The nonglycosylated low M(r) form of the enzyme was not secreted as shown by the decreasing activity in cell-free medium which paralleled the time-dependent secretion of the enzyme. Tunicamycin had no effect on BSDL synthesis. Nevertheless, the nonglycosylated BSDL was apparently not degraded in any cell compartment as shown in part by the enzyme activity accumulation within cells upon brefeldin A treatment. The BSDL expressed by AR 4-2 J cells was characterized by a Km of 68 +/- 30 microM and a kcat of 106 +/- 19 min-1. The sequestrated BSDL due to tunicamycin treatment of cells presents a significant increase of Km of over 10 times to 757 +/- 303 microM. kcat was affected by a factor of approximately 4-445 +/- 22 min-1. These data correlated with an approximately 2.5-fold decrease of the esterolytic activity following inhibition of the N-glycosylation of the protein. The nonglycosylated BSDL was less stable to temperature than the native form. Processing inhibitors (castanospermine, 1 deoxymannojirimycin, and swainsonine) had no effect either on the enzyme activity or on its secretion. Results suggested first that the transfer of the oligosaccharide precursor to the nascent BSDL is essential for the folding of the fully active BSDL. Second that the glycan structure is required for the enzyme secretion. PMID- 8245012 TI - Lysophosphatidylcholine reversibly arrests exocytosis and viral fusion at a stage between triggering and membrane merger. AB - Little is known of the events occurring between membrane fusion triggering and subsequent fusion steps. To dissect this process we applied a reversible inhibitor of membrane fusion, lysophosphatidylcholine, to arrest exocytosis and virus-mediated syncytia formation. Next Ca2+ or H+ (the respective fusion triggers) was administered and later removed. Then, inhibitor was withdrawn and fusion ensued, demonstrating that triggering causes the formation of an "activated state," which later develops into the fused state. Therefore, while different fusion processes utilize different triggers, the pivotal step involving membrane merger is trigger-independent and lipid-sensitive. PMID- 8245013 TI - The M(r) 35,000 beta-adrenergic receptor mRNA-binding protein induced by agonists requires both an AUUUA pentamer and U-rich domains for RNA recognition. AB - Delineating the molecular basis for agonist-induced destabilization of mRNA of G protein-linked receptors that contributes to receptor down-regulation is fundamental to our understanding of long-term regulation of receptors by agonist. Previously we identified a prominent, M(r) 35,000 cytosolic RNA-binding protein that (i) binds selectively to beta 1 and beta 2-adrenergic receptor mRNAs, both of which undergo agonist-induced down-regulation; (ii) does not bind either to alpha 1b-adrenergic receptor mRNA, which does not undergo agonist-induced down regulation, or to beta-globin mRNA; (iii) displays binding to beta 2-adrenergic receptor mRNA that is selectively competed by poly(U) RNA, but not poly(A),-(C), or -(G) RNA; and (iv) its abundance varies inversely with the level of receptor mRNA, being induced by agonists that down-regulate receptor mRNA (Port, J. D., Huang, L.-y., and Malbon (1992) J. Biol. Chem. 267, 24103-24108). We demonstrate here that the binding of beta-adrenergic receptor mRNA by this protein, termed beta-ARB protein, is sensitive to competition by AU-rich domains of the 3' untranslated regions of c-fos, c-myc, and human granulocyte-macrophage colony stimulating factor. Using the AU-rich 3'-untranslated regions of wild-type adenovirus IVa2 mRNA and variants with defined mutations in the AUUUApentamer, AU rich, and U-rich domains, we were able to define sequences critical to the binding of the beta 2-receptor mRNA by the beta-ARB protein. Recognition of beta ARB protein requires not only an AUUUA destabilization pentamer, but also a flanking U-rich domain(s). Using radiolabeled 3'-untranslated regions of short lived mRNA, we were able to identify this same M(r) 35,000 cytosolic RNA-binding protein(s), beta-ARB protein, as selective for beta 2-adrenergic receptor mRNA. PMID- 8245014 TI - Signaling-defective mutants of the B lymphocyte antigen receptor fail to associate with Ig-alpha and Ig-beta/gamma. AB - The B cell receptor for antigen, the membrane immunoglobulin (mIg) molecule, is normally expressed in association with at least two other proteins, mb-1 (Ig alpha) and B29 (Ig-beta/gamma). We have previously described a set of murine B cell clones transfected with wild-type and mutated forms of membrane-bound IgM (mIgM) that vary in their ability to transduce signals to the cell interior. Analysis of the B cell receptor complex in signaling and nonsignaling mutants of mIgM reveals complete concordance between intact signaling ability and ability of the mIgM to associate with Ig-alpha and Ig-beta/gamma. These results point to the importance of the mIgM transmembrane region in mediating binding to accessory molecules, and provide support for a role of Ig-alpha and Ig-beta/gamma that extends beyond surface expression to signal transduction in B lymphocytes. PMID- 8245015 TI - A serine phosphorylation of Nm23, and not its nucleoside diphosphate kinase activity, correlates with suppression of tumor metastatic potential. AB - We describe a serine phosphorylation of the putative metastasis suppressor protein Nm23, and present evidence of its relevance to the signal transduction and tumor metastatic processes. Nm23 was previously demonstrated to exhibit nucleoside diphosphate kinase (NDPK) activity, which transfers a phosphate among nucleoside tri- and diphosphates via an Nm23-phospho-histidine intermediate. Recent data have dissociated the NDPK activity of Nm23 from its phenotypic effects; therefore we have asked whether Nm23 possesses additional biochemical functions. An acid-stable (nonhistidine) phosphorylation was identified on autophosphorylated purified recombinant Nm23 proteins and [32P]orthophosphate labeled human breast carcinoma and murine melanoma Nm23. Phosphoamino acid analysis identified serine as the acid-stable phosphorylation and serine 44 as the major site of phosphorylation. The acid stable phosphorylation (serine) of Nm23 was inhibited by cAMP in vitro and forskolin in vivo, suggesting that this phosphorylation pathway is regulated in signal transduction. No effect of cAMP was observed on Nm23 NDPK activity. Once phosphorylated, Nm23-phosphoserine can release free phosphate in vitro. The biological relevance of the novel phosphorylation identified herein is suggested by the direct correlation of in vivo Nm23 acid-stable phosphorylation levels, but not Nm23 NDPK activity, with suppression of tumor metastatic potential among control and nm23-1 transfected murine melanoma cells. PMID- 8245016 TI - Specific interaction of SPARC with endothelial cells is mediated through a carboxyl-terminal sequence containing a calcium-binding EF hand. AB - SPARC is a secreted, Ca(2+)-binding protein that modulates cell shape and gene expression (Sage, E.H., and Bornstein, P. (1991) J. Biol. Chem. 266, 14831 14834). In the present study we questioned whether SPARC interacted with an endothelial cell surface receptor. The binding of 125I-SPARC to bovine aortic endothelial cells was dependent on Ca2+ and was sensitive to small changes in extracellular pH; maximal binding occurred at pH 7.1. Scatchard analysis indicated approximately 2.3 x 10(7) binding sites/cell with an apparent KI of 1.1 nM. The interaction was diminished specifically by competition with synthetic peptides corresponding to amino acids 54-73 (SPARC 54-73) and 254-273 (SPARC254 273). The binding of 125I-SPARC254-273, a sequence containing a Ca(2+)-binding EF hand, was saturated within 45 min at a concentration of 5 microM; Scatchard analysis indicated 4.2 x 10(7) sites/cell and a KI of 2.4 nM. Iodinated proteins from plasma membranes were affinity-chromatographed on SPARC254-273; several proteins with apparent masses ranging from 153 to 100 kDa (unreduced) or from 153 to 122 kDa (reduced) were eluted with the soluble peptide. These proteins represent candidates for a SPARC receptor(s) that mediates the biological activity of this protein on endothelial cells. PMID- 8245017 TI - Human cartilage gp-39, a major secretory product of articular chondrocytes and synovial cells, is a mammalian member of a chitinase protein family. AB - One of the major secreted proteins of human articular chondrocytes in monolayer or explant culture and of synovial fibroblasts is a glycoprotein with an apparent molecular weight of approximately 39,000, referred to as human cartilage glycoprotein-39 (HC gp-39). The protein was purified, and its complete cDNA sequence was determined. It contained an open reading frame coding for a 383 amino acid long peptide. Comparison of the deduced amino acid sequence with known sequences revealed that HC gp-39 contained regions displaying significant homology with a group of bacterial and fungal chitinases and a similar enzyme found in the nematode, Brugia malayi. In addition significant homologies were observed with three mammalian secretory proteins of as yet unknown function, suggesting that a related protein family exists in mammals. The human protein does not possess any glycosidic activity against chitinase substrates, arguing against any function as an endoglycosidase with specificity for N acetylglucosamine. Analysis by Northern blotting and by reverse transcription/polymerase chain reaction showed mRNA for HC gp-39 to be present in human articular chondrocytes as well is in liver, while mRNA was undetectable in muscle tissues, lung, pancreas, mononuclear cells, or fibroblasts. Neither the protein nor mRNA for HC gp-39 was detectable in normal newborn or adult human articular cartilage obtained at surgery, while mRNA for HC gp-39 was detectable both in synovial specimens and in cartilage obtained from patients with rheumatoid arthritis. These observations suggest that the expression of HC gp-39 may be related to a response of these cells to an altered tissue environment. PMID- 8245018 TI - Mutation of histidine 373 to leucine in cytochrome P450c17 causes 17 alpha hydroxylase deficiency. AB - We identified a new homozygous missense mutation His373-->Leu in the CYP17 gene of two sisters with 17 alpha-hydroxylase deficiency with an elevated plasma aldosterone concentration by sequencing their genomic DNAs amplified by polymerase chain reaction. Using polymerase chain reaction-based site-directed mutagenesis, we prepared a DNA that encoded the Leu373 mutant protein. COS-1 cells transfected with the mutant DNA, despite having an RNA hybridizable to the P450c17 cDNA, did not show 17 alpha-hydroxylase and 17,20-lyase activities. Also, the cells were devoid of 11 beta-hydroxylase and aldosterone synthase activities. To examine the mechanism by which the single amino acid change His373-->Leu eliminates activity, we expressed N-terminally modified P450c17 proteins with and without the Leu373 mutation in Escherichia coli and performed spectral studies. Membrane preparations from E. coli cells expressing the wild-type form of the modified enzyme showed an absorption peak at 449 nm upon addition of carbon monoxide in the reduced state and produced characteristic substrate-induced difference spectra, whereas those from the cells expressing the mutant form did not show these spectral changes. The 17 alpha-hydroxylase and 17,20-lyase activities were observed only in E. coli cells expressing the wild-type enzyme. These results show that the His373-->Leu mutant does not incorporate the heme prosthetic group properly and suggest a critical role of His373 in heme binding. PMID- 8245019 TI - Possible role of both the alpha and beta gamma subunits of the heterotrimeric G protein, Gs, in transcytosis of the polymeric immunoglobulin receptor. AB - In endosomes the polymeric immunoglobulin receptor (pIgR) is sorted into transcytotic vesicles. Transcytosis of the pIgR in polarized Madin-Darby canine kidney (MDCK) cells is regulated both by phosphorylation of Ser664 and by binding of ligand, dimeric IgA (dIgA). Mutation of Ser664 to Ala (pIgR-Ala664) greatly reduces transcytosis of the pIgR without dIgA bound. Here we report that cholera toxin, which specifically activates the heterotrimeric G protein, Gs, stimulates transcytosis of pIgR-Ala664 when dIgA is not bound. However, transcytosis of the wild-type pIgR without dIgA bound was not stimulated by cholera toxin. To further analyze the involvement of Gs in transcytosis, we reconstituted part of the transcytotic pathway. Vesicles containing endocytosed ligand bound to the pIgR could be released from MDCK cells whose apical surface had been mechanically perforated. Release of these vesicles was controlled by phosphorylation of the pIgR and by binding of dIgA. The use of perforated cells allowed us to directly address the involvement of Gs. Our data suggest that both subunits of Gs, Gs alpha and beta gamma, function together to control vesicular release and may therefore be involved in transcytosis in vivo. PMID- 8245020 TI - Purification and characterization of UDP-N-acetylgalactosamine GM3/GD3 N acetylgalactosaminyltransferase from mouse liver. AB - A UDP-N-acetylgalactosamine: Sia alpha 2-3Gal beta 1-4Glc beta 1-/Sia alpha 2 8Sia alpha 2-3Gal beta 1-4Glc beta 1-1ceramide N-acetylgalactosaminyltransferase has been purified to apparent homogeneity from mouse liver. The purification procedure involved differential centrifugation for preparation of Golgi membranes, extraction of the enzyme with Triton X-100, and sequential chromatography on phosphocellulose, UDP-aldehyde adipic acid hydrazone agarose, UDP-hexanolamine-Sepharose, CM-Sepharose, and DEAE-Sepharose. At the phosphocellulose column chromatography step, the recovery of the enzyme activity was less than 25%, but it was enhanced up to 70% when the enzyme assay was performed in the presence of the flow-through fraction from the phosphocellulose column. With this assay, the enzyme activity was found to be quantitatively recovered during all the column chromatographies, the enzyme finally being purified 171,000-fold with a specific activity of 3.6 mumol/min/mg protein. The apparent molecular mass of the purified enzyme is 65,000 daltons. The enzyme exhibits a pH optimum of 7.5-7.9 and requires 2.5-10 mM Mn2+ for the maximal activity. The Km value for UDP-N-acetylgalactosamine is 7 microM. Among the glycolipids tested as acceptor substrates, NeuGc alpha 2-3Gal beta 1-4Glc beta 1 1ceramide, NeuAc alpha 2-3 Gal beta 1-4Glc beta 1-1ceramide, NeuGc alpha 2-8NeuGc alpha 2-3Gal beta 1-4Glc beta 1-1ceramide, and NeuAc alpha 2-8NeuAc alpha 2-3Gal beta 1-4Glc beta 1-1ceramide are good ones, the Km values for them being 160, 2,100, 27, and 350 microM, respectively, but sialyllactose, NeuAc alpha 2-3Gal beta 1-4Glc, is not. This suggests that the enzyme recognizes not only the oligosaccharide portion but also the ceramide of gangliosides. PMID- 8245021 TI - Expression and function of the cytoplasmic variants of the integrin alpha 6 subunit in transfected K562 cells. Activation-dependent adhesion and interaction with isoforms of laminin. AB - Two variants of the cytoplasmic domain of the integrin alpha 6 subunit have been identified (alpha 6A and alpha 6B). To determine the role of each variant in mediating cell adhesion to laminin, we have independently expressed the alpha 6A and alpha 6B subunits in K562 cells. Both variants associated with endogenous beta 1 and were present at comparable levels on the surface of transfected K562 cells. After activation with phorbol ester (phorbol 12-myristate 13-acetate; PMA) or the stimulatory anti-beta 1 antibody TS2/16, alpha 6A beta 1 as well as alpha 6B beta 1 mediated cell adhesion to laminin and more specifically to its fragment E8. Furthermore, both integrin variants interacted with the laminin isoforms kalinin and merosin. Cell adhesion to laminin isoforms was inhibited by the alpha 6-specific monoclonal antibody GoH3. PMA was less efficient in stimulating adhesion than TS2/16 and stimulated adhesion of alpha 6B transfectants better than of alpha 6A transfectants. In contrast, TS2/16 stimulated the adhesion of the alpha 6A and alpha 6B transfectants to laminin to a similar extent. These findings indicate that the cells may regulate the activation of the two alpha 6 variants independently. Activation by PMA was associated with the phosphorylation of both alpha 6A and alpha 6B subunits, but there was no relationship between the degree of phosphorylation and the ability of the transfectants to adhere to laminin since alpha 6A became phosphorylated much more strongly by PMA than alpha 6B. Thus, both alpha 6A beta 1 and alpha 6B beta 1 on K562 cells are activation dependent receptors for different isoforms of laminin. PMID- 8245022 TI - The X-ray structure of Synechococcus ribulose-bisphosphate carboxylase/oxygenase activated quaternary complex at 2.2-A resolution. AB - The structure of the hexadecameric ribulose-bisphosphate carboxylase/oxygenase from Synechococcus PCC6301 has been solved to 2.2-A resolution. Crystallization was in the presence of CO2, Mg2+, and 2'-carboxyarabinitol bisphosphate to form a stable enzyme quaternary complex that mimics one of the intermediate states of the carboxylation reaction. The structure was solved by molecular replacement using the coordinates of spinach carboxylase. The deviations in C alpha positions of the L- and S-subunits are only 0.3 and 2.0 A, respectively, and localized at specific regions of the two polypeptides. One region that shows significant divergence of the peptide backbone is loop 6 of the beta barrel in the L-subunit. Two other elements, the C terminus, and a highly conserved loop of the N-terminal domain of a second L-subunit, interact with loop 6 in the quaternary complex. These three regions, plus two other flexible segments, completely enfold the bisphosphate inhibitor. Significant alteration in their spatial relationship must occur to allow substrates or products access to and from the active site. The active site residues, activating cofactors, and inhibitor are well resolved in the electron density map. The disposition of these groups around the essential metal provides some indication of their role at different stages of the catalytic cycle. PMID- 8245023 TI - Inositol 1,4,5-trisphosphate-mediated quantal Ca2+ release measured by high resolution imaging of Ca2+ within organelles. AB - The distribution and operation of Ca2+ pools within cells has been directly studied in situ by monitoring the Ca2+ inside Ca2+ dye-loaded organelles using high resolution imaging procedures. Using DDT1MF-2 smooth muscle cells, loaded with fura-2 under conditions favoring dye entry into organelles and subjected to carefully controlled permeabilization still attached to coverslips, the Ca2+ within organelles was analyzed by high resolution, z axis-controlled imaging, and deblurring methods. Saturation analysis of entrapped fura-2 indicated that the dye reported Ca2+ identically to fura-2 in solution. Areas containing high Ca(2+) sequestering organelles (> 5 microM free Ca2+) were observed to predominate around the nucleus and close to the periphery of the cell. Analysis of the actions of inositol 1,4,5-trisphosphate (InsP3) within small (3 microns 2) selected intracellular areas, revealed a "quantal" release phenomenon, with rapid attainment of limited stable release at submaximal InsP3 levels. The apparent EC50 for InsP3 was approximately 3 microns, higher than within suspensions of permeabilized cells. The action of InsP3 was competitively blocked by 10 micrograms/ml of the InsP3 antagonist, heparin. Applied after maximal InsP3 mediated Ca2+ release, heparin reversed InsP3-induced Ca2+ release resulting in reuptake of Ca2+ into Ca(2+)-pumping organelles with identical spatial distribution as before Ca2+ release. InsP3 released Ca2+ from all areas of high Ca(2+)-pumping organelles; extensive areas of high fura-2-loading, but low intraorganelle Ca2+, were unchanged by InsP3. GTP induced no alteration in Ca2+ release (in contrast to suspensions of permeabilized cells), suggesting that the InsP3-sensitive Ca2+ pool was functioning as a single homogeneous pool. Opening of InsP3-sensitive channels was also monitored by assessing InsP3-activated channel-mediated Mn2+ quenching of organelle-loaded fura-2; the results revealed a similar pattern of quantal release, with slightly increased apparent InsP3 sensitivity. The results provide the first high resolution in situ localization of Ca2+ signaling organelles and demonstrate the quantal operation of InsP3 sensitive Ca2+ pools within highly discrete subcellular loci. PMID- 8245024 TI - Alteration of splice site selection by an exon mutation in the human glycophorin A gene. AB - We report the identification in human glycophorin A gene of a novel exon mutation that affects splice site selection. DNA mapping showed that the mutation has abolished a unique MspI site marking the exon III-intron 3 splice junction (ACCG/GT). Genomic sequencing confirmed the occurrence of a single G-->A transition at the terminal nucleotide position of exon III. Analysis of the mRNA composition demonstrated a partial inactivation of the altered 5' splice site as well as skipping of exons involving the alternative use of other constitutive splice sites. The full-length transcript with the cognate A change encodes a variant glycophorin with an arginine replacing a glycine at position 59 and defining the ERIK epitope, whereas the exon III-deleted transcript specifies a shorter glycophorin carrying the Sta antigen. Also identified were the misspliced mRNA species with exon I-IV and exon I-V connections generated by selection of 5' splice sites far distant from the mutated site. Although having a correct translation frame, the predicted polypeptides of such exon-skipping products were not assembled on the erythrocyte membrane probably due to the severe truncation of the signal sequence required for targeting and translocation. These observations reveal a new mechanism for antigenic diversity of human glycophorins. PMID- 8245025 TI - Crystal structures of oxidized and reduced Azotobacter vinelandii ferredoxin at pH 8 and 6. AB - Crystal structures of Azotobacter vinelandii ferredoxin I (FdI) have been solved and refined at 2.2 to 1.9-A resolution at pH 8 and 6 for both the oxidized and dithionite-reduced proteins. Only the [3Fe-4S] cluster is reduced by dithionite. The four structures (denoted FdI8ox, FdI8red, FdI6ox, and FdI6red) have been compared to address three questions: the effect of reduction at pH 8, the effect of pH change on the structure, and the effect of reduction at pH 6. Comparison of the FdI8ox and FdI8red structures shows that Asp-15 changes conformation in a manner consistent with increased anionic repulsion between this residue and the reduced [3Fe-4S]0 cluster. By revealing an electrostatic interaction between Asp 15 and the [3Fe-4S] cluster, this result supports the conclusion in the accompanying paper (Shen, B., Martin, L. L., Butt, J. N., Armstrong, F. A., Stout, C. D., Jensen, G. M., Stephens, P. J., LaMar, G. N., Gorst, C. M., and Burgess, B. K. (1993) J. Biol. Chem. 268, 25928-25939) that Asp-15 participates in protonation of the reduced [3Fe-4S]0 cluster at acid pH. The [3Fe-4S]0 cluster in the FdI8red structure also displays a distinct shift within the protein as well as internal distortions when compared to the [3Fe-4S]+ cluster in the FdI8ox structure. Comparison of the FdI8ox and FdI6ox structures shows that pH change does not have any significant effect on the [3Fe-4S]+ cluster or surrounding residues. Comparison of the FdI6ox and FdI6red structures shows that reduction at pH 6 also does not have any significant effect on the [3Fe-4S] cluster or Asp-15. The absence of structural change supports the conclusion that at acid pH, the reduced [3Fe-4S] cluster is protonated, i.e. [3Fe-4S]0-H+ (Shen et al., 1993). The cluster is not shifted or distorted as in the FdI8red structure. Instead, the [3Fe-4S]o-H+ cluster FdI8red is structurally similar to the [3Fe-4S]+ cluster (FdI8ox, FdI6ox), which has the same net charge. An Asp-15-Lys-84 salt bridge is observed in all four structures, indicating that Asp-15 is ionized at pH 8 and 6. An ionized state for Asp-15 is also implied by a lack of conformational change in Lys-84; the side chain of this residue rearranges when Asp-15 is substituted with a neutral amino acid (Shen et al., 1993).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8245026 TI - Azotobacter vinelandii ferredoxin I. Aspartate 15 facilitates proton transfer to the reduced [3Fe-4S] cluster. AB - The [3Fe-4S]+/0 cluster of Azotobacter vinelandii ferredoxin I (AvFdI) has an unusually low and strongly pH-dependent reduction potential (E'0). The reduced cluster exists in two forms, depending upon pH, that exhibit substantially different magnetic circular dichroism (MCD) spectra. Recent studies have established that the MCD changes observed on decreasing the pH from 8.3 (alkaline form) to 6.0 (acid form) cannot be explained either by a change in spin state of the cluster (Stephens, P.J., Jensen, G.M., Devlin, F.J., Morgan, T.V., Stout, C. D., Martin, A.E., and Burgess, B.K. (1991) Biochemistry 30, 3200-3209) or by a major structural change (e.g. ligand exchange) (Stout, C.D. (1993) J. Biol. Chem. 268, 25920-25927). Here, we have examined the influence of aspartate 15 on the pH dependence of the spectroscopic and electrochemical properties of AvFdI by construction of a D15N mutant. Aspartate 15, which is salt-bridged to lysine 84 at the protein surface, is the closest ionizable residue to the [3Fe-4S] cluster. The results show that replacement of aspartate by asparagine results in an approximately 20-mV increase in E'0 for the [3Fe-4S]+/0 cluster at high pH concomitant with an approximately 0.8-pH unit decrease in the pK of the reduced form. The major pH dependence of E'0 is preserved as is the effect observed by MCD. These data eliminate the possibility that the MCD change is due to the presence of Asp-15 and support the conclusion that it originates in direct protonation of the [3Fe-4S]0 cluster, probably on a sulfide ion. Voltammetric studies show that interconversion between [3Fe-4S]+ and [3Fe-4S]0 at acidic pH involves rapid electron transfer followed by proton transfer (for reduction) and then proton transfer followed by electron transfer (for oxidation). Ionized aspartate 15 facilitates proton transfer. Thus, protonation and deprotonation are much slower for D15N relative to the native protein at pH > 5.5. Proton transfer reactions necessary for further reduction of the [3Fe-4S]0 cluster to the [3Fe 4S]- and [3Fe-4S]2- states are also retarded in D15N. The results suggest that the carboxylate-ammonium salt bridge afforded by Asp-15-Lys-84 conducts protons between the cluster and solvent H2O molecules. Overproduction of D15N FdI, but not native FdI, in A. vinelandii has a negative effect on the growth rate of the organism, suggesting that the rate of protonation or deprotonation of the [3Fe 4S]0 cluster may be important in vivo. PMID- 8245027 TI - Rapid degradation of an unassembled immunoglobulin light chain is mediated by a serine protease and occurs in a pre-Golgi compartment. AB - The immunoglobulin kappa light chain produced by the CH12 lymphoma is unusual because it is not secreted when expressed in the absence of a heavy chain. Instead, it undergoes rapid intracellular degradation. This degradation is selective, as another light chain expressed in the same cell is not degraded. It is also a property of the CH12 kappa chain itself, since it is degraded rapidly when expressed either in another myeloma cell or in COS-1 fibroblasts. When provided a heavy chain, this kappa chain assembles into IgM and is then protected from proteolysis. The degradation of kappa requires ATP, is sensitive to reduced temperature and to the thiol reagent diamide. Of all the proteolytic inhibitors tested, 3,4-dichloroisocoumarin, L-1-tosylamido-2-phenylethyl chloromethyl ketone, and to a lesser extent 1-chloro-3-tosylamido-7-amino-2-heptanone, inhibit kappa degradation, suggesting the involvement of a serine protease. The degradation of kappa does not require transport to the Golgi complex, nor is it sensitive to a variety of lysosomotropic agents. Both immunofluorescence and the observed association with the endoplasmic reticulum (ER) stress proteins GRP78/BiP and GRP94 indicate that the kappa chain is localized mostly in the ER. When a point mutation which blocks transport to the Golgi complex is introduced into this kappa chain, the association with the stress proteins is enhanced but the rate of degradation is not significantly decreased. We conclude that the CH12 kappa chain is a particularly good substrate for an ER degradation machinery, and that its sensitivity to the protease(s) is governed by its state of assembly. This ER degradation provides a possible quality control mechanism during the differentiation of B lymphocytes. PMID- 8245028 TI - Activation of phospholipase C-beta 2 mutants by G protein alpha q and beta gamma subunits. AB - The beta- but not the gamma- and delta-type isozymes of inositol phospholipid specific phospholipase C (PLC) are activated by G protein alpha q and beta gamma subunits. The beta-type PLC isozymes differ from other isozymes in that they contain a long carboxyl-terminal region downstream of the Y catalytic domain and a region rich in acidic amino acids between the two separated X and Y catalytic domains. To determine the sites on PLC-beta 2 that participate in the interaction of the enzyme with alpha q and beta gamma subunits, we introduced specific truncations and substitutions in the PLC-beta 2 cDNA at positions corresponding to the carboxyl-terminal and acidic amino acid-rich regions, respectively. After transient expression of these cDNA clones in CV-1 cells, the mutant enzymes were partially purified and their capacity to be activated by alpha q and beta gamma subunits determined. Substitution of glutamine residues for three or all seven of a stretch of consecutive glutamic acids in the acidic domain of PLC-beta 2 affected neither alpha q- nor beta gamma-dependent activation significantly. Carboxyl-terminal truncation to residue Gly-934 or to residue Ala-867 resulted in enzymes that were activated by beta gamma but not by alpha q. This result suggests that the carboxyl-terminal region of PLC-beta 2 is required for activation by alpha q, and that beta gamma subunits interact with a different region of the enzyme. Thus, alpha q and beta gamma subunits may independently modulate a single PLC-beta 2 molecule concurrently. PMID- 8245029 TI - Targeting and translocation of the phosphate carrier/p32 to the inner membrane of yeast mitochondria. AB - We analyzed the submitochondrial location and biogenesis pathway of the phosphate carrier (PiC), also termed p32, of Saccharomyces cerevisiae mitochondria, PiC/p32 was found to behave as an integral membrane protein that cofractionated with the ADP/ATP carrier of the inner membrane. Import of the precursor of PiC/p32 required a membrane potential across the inner membrane, supporting its localization to the inner membrane. This makes it unlikely that the major function of PiC/p32 is that of an import receptor on the surface of the mitochondrial outer membrane. Furthermore, we found that both receptors MOM72 and MOM19 were involved in the import pathway of the precursor of PiC/p32 with MOM72 being responsible for the bulk of import. Yeast PiC/p32 is thus not only structurally homologous to the ADP/ATP carrier, but has a similar targeting mechanism and submitochondrial location, supporting its classification as a member of the inner membrane carrier family. PMID- 8245030 TI - Hysteresis in the triple helix-coil transition of type III collagen. AB - A hysteresis is observed in the triple helix-coil transition of type III collagen. This hysteresis appears in the thermal transition, where it is not solvent-dependent, and also in the transition induced by guanidinium hydrochloride. The transitions are observed both by optical rotatory dispersion and by trypsin digestion. Experiments with various fragments of type III collagen, which, like the intact protein, contain a set of disulfide bonds at the carboxyl-terminal end, which functions as an effective nucleus for refolding, indicate that the hysteresis depends on the amino acid sequence, rather than being a general property of the triple helix. The carboxyl-terminal one-quarter fragment of type III collagen does not show a hysteresis, whereas a mutant type III collagen missing residues 595-1008 shows a hysteresis of the same magnitude as normal type III collagen. Several fragments were produced that are missing parts of the amino-terminal end, and these fragments show no hysteresis. The amino-terminal part of the triple helix of type III collagen consists of a stable region containing six Gly-Pro-Hyp tripeptide units within the first 16 tripeptide units. This region forms an amino-terminal clamp for the triple helix and, together with the set of disulfide bonds at the carboxyl-terminal end, allows for highly cooperative unfolding of the triple helix. The presence of this amino terminal region is a prerequisite for the observed hysteresis. PMID- 8245031 TI - Involvement of mitochondrial contact sites in the subcellular compartmentalization of phospholipid biosynthetic enzymes. AB - The concerted synthesis of phospholipids derived from serine involving two microsomal enzymes (phosphatidylserine synthase and phosphatidylethanolamine N methyltransferase) and a mitochondrial one (phosphatidylserine decarboxylase) occurs in reconstituted cell-free systems. Subfractionation of crude mitochondria after swelling and separating on a sucrose density gradient resulted in the isolation of two contact site-enriched fractions from total outer membranes and inner membranes, respectively. Estimation of marker enzyme activities shows a high recovery of glucose-6-phosphate phosphatase (a marker for the endoplasmic reticulum) associated with contact site-enriched fractions. Accordingly, the linked synthesis of phosphatidylserine, phosphatidylethanolamine, and at a lesser extent phosphatidylcholine can occur. This biosynthetic pathway was absent from purified contact site-enriched fractions correlative with the absence of glucose 6-phosphate phosphatase activity. Reconstitution experiments, including contact site-enriched fractions incubated with endoplasmic reticulum-rich fraction, led to the restoration of the linked synthesis of phospholipids, thereby demonstrating that a reversible association between these two fractions can occur. These functional interactions between the endoplasmic reticulum and mitochondria are confirmed at the ultrastructural level using either chemical or physical fixation before resin embedding. These results show that the interorganelle trafficking of lipids may involve only highly specialized microdomains of both membranes, thereby allowing the maintenance of a specific lipid composition and distribution within membranes. PMID- 8245032 TI - Quantitative analysis of alternative splicing options of human plasma membrane calcium pump genes. AB - The alternative splicing options and the quantitative tissue distribution of the transcripts of the four currently known human plasma membrane calcium pump (PMCA) genes have been analyzed in seven tissues (cerebral cortex, skeletal and heart muscle, stomach, liver, lung, and kidney) by quantitative polymerase chain reaction on reverse transcribed mRNA with glyceraldehyde-3-phosphate dehydrogenase as the internal standard. The mRNAs of genes 1 and 4 were found to be present in similar amounts in all tissues, whereas the transcripts of genes 2 and 3 were expressed in a tissue-specific manner, i.e. their amounts were highest in fetal skeletal muscle and brain. Alternative splicing was found to occur in the PMCA transcripts at two major regulatory sites (sites A and C), adjacent to the amino-terminal phospholipid-responsive region and within the carboxyl terminal calmodulin binding domain, respectively. Novel splicing variants not described previously for human genes were detected for hPMCA3 and 4 at site A and for hPMCA1, 2, and 3 at site C. For all genes a common splice variant was found at both splice sites. The common splice variant at site A was characterized by the inclusion of a small exon (hPMCA1, 39 base pairs (bp); hPMCA2, 42 bp; hPMCA3, 42 bp; hPMCA4, 36 bp). In the common splice variant at site C, an exon (hPMCA1, 154 bp; hPMCA2, 227 bp; hPMCA3, 154 bp; hPMCA4, 178 bp) was excluded in the mRNA. All genes normally express these main splice variants in all tissues in which the corresponding isoform is present. The splicing complexity at site C was found to be augmented in the transcripts of PMCA2 and PMCA3 through the use of additional exons, and in PMCA1 and 3 through the use of additional internal splice sites in the single alternatively spliced 154-base pair exon. PMID- 8245033 TI - Identification of Gq as one of the G-proteins which copurify with human platelet thromboxane A2/prostaglandin H2 receptors. AB - The present study employed ligand affinity and immunoaffinity chromatography to isolate human platelet thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptor coupled G-proteins. Purification of TXA2/PGH2 receptors by ligand (SQ31,491) affinity chromatography resulted in the elution of receptor binding and GTPase activity in the same fraction. GTPase activity of this fraction was enriched (6 fold) relative to solubilized platelet membranes, was stimulated (65%) by 500 nM U46619, and was blocked (74%) by 250 nM SQ29,548. Furthermore, GTP (100 microM) increased [3H]SQ29,548 receptor binding by 48%. Immunoblotting of this fraction against QL antiserum identified a 42-kDa protein as a member of the Gq family. In separate experiments, TXA2/PGH2 receptors were purified by immunoaffinity chromatography using P1Ab, P2Ab, and TxAb affinity columns. QL-immunoreactive proteins at 42 kDa were found in all three column eluates. Studies using G alpha,common antiserum (GA/1) demonstrated immunoblotting of two proteins of approximately 42 and 85 kDa in both the ligand and P2Ab affinity column fractions. On the other hand, the P1Ab and TxAb affinity column eluates contained GA/1 immunoreactivity only in the 42 kDa region. Collectively, these data identify Gq as a TXA2/PGH2 receptor-coupled G-protein and suggest the association of this receptor with additional G alpha subunits. PMID- 8245034 TI - Structural and functional characterization of the HPV16 E7 protein expressed in bacteria. AB - The E7 gene of the human papillomaviruses (HPV) encodes a 98-amino acid, multifunctional nuclear phosphoprotein with functional and structural similarities to adenovirus E1A and the papovavirus T antigens. E7 is a viral oncoprotein, which will cooperate with an activated ras oncogene to transform primary rodent cells, and can cooperate with the HPV E6 protein for the efficient immortalization of primary human keratinocytes. Due to the compelling epidemiological and experimental association between HPV infection and cervical cancer, we have undertaken a detailed study of the structure of the HPV16 E7 protein. The E7 protein was expressed in Escherichia coli as a native, unfused polypeptide, and soluble protein was purified by conventional chromatographic techniques. The purified protein was assessed for various biochemical and biophysical properties. Purified E7 binds the retinoblastoma protein avidly and specifically, and it can dissociate the E2F transcription factor when assayed in vitro. Circular dichroism spectroscopy indicated that E7 reversibly binds Zn2+ and Cd2+, resulting in a substantial increase in the alpha-helical content of the metal-bound E7 consistent with the stabilization of a hydrophobic core in the COOH terminus of the protein. PMID- 8245035 TI - Goniodomin A induces modulation of actomyosin ATPase activity mediated through conformational change of actin. AB - Goniodomin A, one of the polyether macrolides, isolated from the dinoflagellate Goniodoma pseudogoniaulax stimulated skeletal actomyosin ATPase activity 2-3 times, with a peak at 3 x 10(-8) to 3 x 10(-7) M. The effect of goniodomin A was dependent on the concentration of actin, but not of myosin. The actomyosin ATPase activity was increased by pretreatment of actin (but not of myosin) with goniodomin A. Goniodomin A induced a sustained and concentration-dependent increase in the fluorescence intensity (excitation wavelength, 277 nm; emission wavelength, 329 nm) of actin. The maximum response was obtained with concentrations of goniodomin A in the 10(-5) to 10(-4) M range in the presence of 5 microM F-actin. However, the ATPase activity and fluorescence intensity of myosin were not changed by goniodomin A at concentrations from 10(-8) to 10(-5) M. Interestingly, goniodomin A induced a remarkable but transient increase in the fluorescence intensity of actomyosin in a concentration-dependent manner, with a peak at 3 x 10(-7) M. This profile was quite similar to that found in the stimulation of the actomyosin ATPase activity induced by goniodomin A. To investigate further the effect of goniodomin A, actin was labeled with N-(1 pyrenyl)iodoacetamide. Goniodomin A at 10(-6) M had no effect on the fluorescence intensity of pyrenyl-actin (excitation wavelength, 365 nm; emission wavelength, 407 nm), but increasing concentrations of goniodomin A to 3 x 10(-6) M remarkably decreased its intensity. This effect was potentiated by heavy meromyosin. Actin molecules treated with goniodomin A were completely sedimented by mild centrifugation (for 15 min at 12,000 x g). Electron microscopic observations suggest that actin filaments associate with each other to form a gel in the presence of 3 x 10(-6) M goniodomin A. The conformational change of actin molecules, resulting from stoichiometric binding of goniodomin A to actin monomers in filaments, may modify the interaction between actin and myosin. PMID- 8245036 TI - Microencapsulated human hepatoma (HepG2) cells: in vitro growth and protein release. AB - The feasibility of a microencapsulation process ultimately for cell transplantation was investigated by encapsulating human hepatoma (HepG2) cells in hydroxyethyl methacrylate-methyl methacrylate (HEMA-MMA) membranes through an interfacial precipitation process. Changes in viability and metabolic activity as well as protein secretion by the encapsulated cells were studied in vitro. When encapsulated at either low or high density (1 or 5 x 10(6) cells/mL, respectively), HepG2 cells retained their active metabolic state and/or proliferated during the initial 1-week period, after which a significant drop in cell viability was obtained. Encapsulation of a biological attachment substrate, Matrigel, along with the cells, however, resulted in rapid proliferation in both low and high density capsules with prolonged maintenance of an active metabolic state. The secretion of four model proteins (alpha 1-acid glycoprotein, alpha 1 antitrypsin, haptaglobin and fibrinogen) was demonstrated during the 2-week study period for the Matrigel encapsulated cells. Furthermore, the encapsulated cells remained responsive to interleukin 6 (IL6), a physiological stimulator of plasma protein secretion, as determined by the elevated secretion of haptaglobin in response to IL6 treatment. We conclude that HEMA-MMA capsules, in the presence of an attachment substrate, provide a suitable environment for the growth and expression of differentiated functions of encapsulated hepatoma cells. PMID- 8245037 TI - Dissolution and scanning electron microscopic studies of Ca,P particle-containing bioactive glasses. AB - Calcium phosphate (Ca,P) precipitation behavior on the surface of two bioactive glasses and four bioactive glass composites--two with hydroxylapatite (Ca10(PO4)6 (OH)2) and two with rhenanite (CaNaPO4)--were studied in simulated body fluid (SBF) and in Tris-Buffer at 5, 8, 16, 24, 48, 72, and 144 h. The weight loss of the materials was measured and the amount of precipitation was estimated using scanning electron microscopy with electrochemical detection (SEM-EDX) analysis. The test was repeated for one glass and its respective rhenanite composite every 3 h until 60 h and thereafter every 10 h until 150 h in SBF. Atomic absorption spectroscopy, spectrophotometry, SEM-EDX analysis, and pH measurements were performed on these samples. It is shown that in vitro the composite materials have a higher capacity for Ca,P precipitation than the glasses. Weight losses of the materials correlate well with their composition. Both the glass and Ca,P phases influence the precipitation mechanism and rate. Precipitation begins preferably from the glass phase. Ca,P particles clearly influence the time of onset and rate of precipitation. Cross-sectional EDX analysis of the samples revealed an absence of a clear Si-rich layer in glass A0B0 (SiO2 53.9 mol %, Na2O 27.5, CaO 12.4, P2O5 6.2, Al2O3 0.0 and B2O3 0.0) composites. This was attributed to the presence of extra calcium and phosphate ions on the surface of the material. The ion-concentration and pH change curves offered insight into the mechanism of precipitation. A connection was established between SEM-EDX results and the release curves. Formation of an Si,Ca,Na film was observed that seemed to initiate the Ca,P precipitation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245038 TI - Directional porosity of porcine small-intestinal submucosa. AB - Small-intestinal submucosa (SIS) has been shown to be a promising biomaterial for vascular graft applications. This study examines the directionality property of SIS porosity using 35 SIS specimens from 13 pigs. In addition, the effects of the weight of the donor pig, pre-conditioning of 13 additional SIS specimens, and the duration of the test of five additional SIS specimens on such porosity are reported. The porosity from serosal to mucosal direction was found to be four times greater than the porosity in the opposite direction. The weight of the donor pig was not found to be an important factor in SIS porosity. Preconditioning served to increase the average serosal porosity index at 120 mm Hg static water pressure from 2.99 to 8.33 mL/(min cm2). The porosity in the mucosal direction was not affected by preconditioning. Porosity in both directions decreased with increasing test duration. The directionality property of SIS porosity may be an important factor in its success as a vascular graft. The term 'porosity' is used throughout this article, but current standards also refer to the term 'permeability' to describe the passage of liquid through a vascular graft. PMID- 8245039 TI - A novel recovery system for cultured cells using plasma-treated polystyrene dishes grafted with poly(N-isopropylacrylamide). AB - Poly(N-isopropyl acrylamide) (PIPAAm) demonstrated a fully expanded chain conformation below 32 degrees C and a collapsed, compact conformation at high temperatures. This unique temperature responsive polymer was grafted onto surfaces of commercial polystyrene dishes and used as temperature switches for creating hydrophilic surfaces below 32 degrees C and hydrophobic surfaces above 32 degrees C. Cell attachment and the growth of bovine endothelial cells and rat hepatocytes on PIPAAm-grafted surfaces at 37 degrees C demonstrated similar behavior to the commercialized culture dishes. Both cell types were observed to detach from the PIPAAm-grafted surface simply by reducing the temperature below the polymer transition temperature (collapse). Cells recovered by this method maintained substrate adhesivity, growth, and secretion activities nearly identical to those found in primary cultured cells in contrast to the compromised function found in cultured cells damaged by trypsinization. These results provide strong evidence that PIPAAm-grafted surfaces, as thermal switches are very effective for reversing cell attachment and detachment without cell damage. Properties of cell culture surfaces can be readily transformed by this technique reversibly into hydrophilic and hydrophobic coatings of PIPAAm-grafted polymers. PMID- 8245040 TI - Poly(ethyleneimine) modified filters for the removal of leukocytes from blood. AB - Polyurethane membrane filters and filters coated with poly(ethyleneimine) were used to investigate the influence of leukocyte adhesion during filtration. Treatment of the filters with an aqueous solution of 1% (w/v) poly(ethyleneimine) (PEI) led to the introduction of amine groups at the filter surfaces, as was confirmed by X-ray photoelectron spectroscopy. The modification procedure did not significantly change the porous structure in the filters, as was demonstrated by SEM and porometry. Using 14C-labeled poly(ethyleneimine) it was shown that nearly a complete coverage (approximately 0.1 mg/m2) was achieved that did not desorb from the filter surface during contact with blood plasma. When the filtration was carried out with purified leukocytes in the absence of red cells, platelets, and blood plasma, the number of cells removed by modified filters (> 95%) was significantly higher as compared to the removal with unmodified filters (approximately 80%). However, no significant differences between the filters were found when the filtration was performed with whole blood. This finding was unexpected, because it was shown before that immobilization of poly(ethyleneimine) on solid polyurethane film, surfaces promoted the adhesion of leukocytes from whole blood. Apparently, the adhesive properties of the PEI diminish during filtration. Filter coating of commercial leukocyte filters composed of polyester fibers also had no effect on the removal of leukocytes from whole blood. It was postulated that morphological factors, such as filter shape, roughness, tortuosity, and porosity rather than the physicochemical properties of the filter surface influence cell adhesion to the filter surface, and through that the filtration process. PMID- 8245041 TI - Compliance effects on small diameter polyurethane graft patency. AB - Microporous compliance matched and noncompliant grafts were compared in a dog carotid artery interposition model. We fabricated 4 mm diameter sponge type polyurethane (Biomer) tubes 5 cm in length with a 0.5 mm wall thickness. The luminal surface was covered with a 50 microns coating of cross-linked gelatin. Compliance was measured in vitro and in vivo by volume and vessel diameter changes. Over a mean arterial pressure range of 55-155 mm Hg, the diameter changes of grafts and stump arteries were measured in situ using an ultrasonic Hokanson device. Compliance matched grafts were found to have the same in vitro compliance values as the natural canine carotid at a mean arterial pressure of 100 mm Hg. Compliance matched and noncompliant grafts had values of 10.3 +/- 1.3 and 0.9 +/- 0.1 x 10(-2) mm Hg, respectively. End to end arterial anastomoses were constructed between the graft and the host arteries. The use of synthetic grafts with matched compliance to the adjacent natural vessels has been advocated as the ideal solution to circumvent the problems of graft failure. These studies indicate that compliance values for compliance matched grafts decreased immediately after implantation (from 10.3 to 6.5 x 10(-2) %/mm Hg) and within 6 weeks decreased to 3.6 x 10(-2) %/mm Hg. The compliance values for noncompliant grafts remained constant throughout the test period. At autopsy all grafts showed a tightly adhered tissue capsule. The thickness of the anastomotic hyperplasia at the distal sites of compliance matched grafts was significantly different (P < .05) than that of the adjacent artery. The patency for compliant and noncompliant grafts was 64% and 50%, respectively. Evidence for polyurethane graft degradation was obtained by Fourier transform infrared spectroscopy and gel permeation chromatography analysis of patent explants. Compliance mismatch alone does not contribute to graft failure, however, material degradation, suture technique and/or capsule formation can play a contributory role although these were not tested directly. PMID- 8245042 TI - Development of FRP composite structural biomaterials: fatigue strength of the fiber/matrix interfacial bond in simulated in vivo environments. AB - Fiber/matrix interfacial bonding in fiber reinforced polymer (FRP) composite materials is potentially sensitive to degradation in aqueous environments. Ultimate bond strength (UBS) in carbon fiber/polysulfone (CF/PSF) and polyaramid/polysulfone (K49/PSF) was previously reported to be significantly decreased in two simulated in vivo environments. While UBS is a useful parameter, for orthopedic implant applications the fatigue behavior of the interface is probably a more relevant indicator of long-term composite material performance. In this article, the effects of simulated in vivo environments (saline, exudate) upon the fatigue behavior of the interface of CF/PSF and K49/PSF are reported. The fatigue behavior of both material combinations was linearly dependent on the logarithm of fatigue life in the dry (control), saline, and exudate environments. Testing either material in saline and exudate resulted in significantly lower fatigue strength than in the dry environment; however, results in the two wet environments were indistinguishable. The CF/PSF interface experienced fatigue failure at approximately 10(5) load cycles at a maximum applied load level of only 15% of its ultimate dry bond strength without indication of an endurance limit being reached. These results raise some important questions regarding the durability of CF/PSF composite in load bearing orthopedic applications. PMID- 8245043 TI - Quantification of the edge effect in calcified bioprosthetic tissues. AB - In bioprosthetic tissue samples that had been implanted in the subcutaneous space of rats, and recurring pattern of calcification was observed. In this pattern, which we call the edge effect, the interior of the tissue is calcified and is surrounded and separated from the subcutaneous fluid by a zone that is free from calcification. The edge effect has been qualitatively described in the literature for subcutaneous implants and for valve leaflets, and it may be related to the mechanism of calcification for these materials. The thickness of the calcification free outer layer was quantified for glutaraldehyde treated bovine pericardium, glycerol treated bovine pericardium, glutaraldehyde treated human dura mater, and glycerol treated human dura mater. The edge effect values were found to be unique and consistent for each material type, and they were inversely related to the shrinkage temperatures and the calcium contents of the materials. It was determined that the chemical treatment was more important than the tissue type in determining the edge effect value. PMID- 8245044 TI - Chemical durability of Y2O3-Al2O3-SiO2 glasses for the in vivo delivery of beta radiation. AB - Microspheres made from Y2O3-Al2O3-SiO2 (YAS) glasses, which contain radioactive Y 90, are currently being used to treat liver cancer in humans, where their chemical durability is of prime importance. In deionized water or saline at 37 degrees C, the weight percent Yttrium (Y) dissolved from eight different YAS glasses ranged from only 0.02-0.13% of the total Y present and their dissolution rate was barely measurable, < or = 1.0 x 10(-9) g/cm2-min. The most chemically durable YAS glass was 17Y2O3-19Al2O3-64SiO2, mol%. The small amount of Y released from microspheres, 25-35 microns diameter, of this glass after corrosion in saline or deionized water at 37 degrees C was essentially the same as for bulk glass samples. Based on their excellent chemical durability, it is concluded that YAS glass microspheres are suitable for in vivo use. PMID- 8245045 TI - Effects of phospholipid adsorption on nonthrombogenicity of polymer with phospholipid polar group. AB - Polymers with phospholipid polar groups, 2-methacryloyloxyethyl phosphorylcholine (MPC) polymers, have excellent nonthrombogenic properties. The effects of adsorption of phospholipids on platelet adhesion and activation on the MPC copolymer with n-butyl methacrylate (BMA) were investigated with particular attention to the structure of the phospholipids adsorbed onto the polymer surface. The electrical nature of the phospholipids adsorbed on the polymer surface affected the thrombogenicity of the polymer. On the MPC polymer surface treated with an aqueous liposomal solution of acidic phospholipids, phosphatidylserine, platelet adhesion and activation occurred to a greater extent when compared to a poly(MPC-co-BMA) surface. However, on the MPC polymer surface treated with electrically neutral phosphatidylcholines, reduced thrombogenicity could be observed. Therefore, the adsorption of the phosphatidylcholines was an important factor in reducing the thrombogenicity on the polymers. Moreover, by comparison of the poly(MPC-co-BMA) to a poly(BMA), platelet adhesion and activation on these polymer surfaces depended on the adsorption state of the phosphatidylcholines. The amount of phosphatidylcholine adsorbed on the poly(MPC co-BMA) increased with an increase in the MPC mole fraction of the copolymer. This indicates that the MPC moieties have affinity for the phosphatidylcholines. We conclude that the poly(MPC-co-BMA) can adsorb large amounts of phosphatidylcholines and that these phospholipids organize themselves. The organized adsorption layer of the phosphatidylcholines on the surface, which construct biomembrane-like surfaces, can reduce platelet adhesion and activation effectively. PMID- 8245046 TI - The shear strength and the failure mode of plasma-sprayed hydroxyapatite coating to bone: the effect of coating thickness. AB - Plasma-sprayed hydroxyapatite coated (HAC) 50 and 200 microns thick on Ti-6Al-4V cylinders was transcortically implanted in the femora of canines to evaluate in detail the effect of coating thickness on the pushout shear strength and failure mode examined under scanning electron microscope after the periods of 4, 6, 8, and 12 weeks. The HAC coating exhibited higher shear strength at 50 microns than at 200 microns. Its failure mode was conclusively at or near the HAC-bone interface, and the slight attack of body fluid had not degraded the implant to the extent that failure occurred at the HAC-Ti alloy interface after 12 weeks of observation. For 200 microns-HAC, failure was found at the HAC-bond interface, inside the HAC lamellar splat layer and at the HAC-Ti alloy substrate interface, depending on the period of implantation. It was also deduced that the variation of failure mode of 200 microns-HAC with time could not be accounted for by the attack of body fluid alone; the degradation must be a synergetic adverse result of residual stress in the HAC and the attack of body fluid. PMID- 8245047 TI - Bicomponent vascular grafts consisting of synthetic absorbable fibers. I. In vitro study. AB - The objective of this study is to determine the effects of the location and concentration of synthetic absorbable yarn components in bicomponent vascular graft fabrics on their structure and properties in a controlled in vitro hydrolytic environment. Bicomponent vascular fabrics were made from Dacron and polyglycolic acid (PGA) yarns with a range of composition ratios of PGA to Dacron and a range of locations of PGA. Both woven and single jersey knit fabrics were made. These fabrics were characterized by standard textile methods and subject to in vitro hydrolytic degradation study. In vitro hydrolytic degradation study showed that the most dramatic changes in the bicomponent fabric characteristics and properties occurred 30 and 60 days of hydrolysis. This schedule coincided with the hydrolytic degradation rate of PGA absorbable sutures. In the woven (W) group, the incorporation of absorbable yarns in the weft direction (W3) of the bicomponent fabrics resulted in the velour-like, loose, and porous surface morphology of the fabric for potential subsequent tissue ingrowth, while those woven fabrics with absorbable yarns in the warp direction (W1) did not have this unique velour-like surface. In the knitted (K) group, the concentration of absorbable yarns appeared to be closely related to the observed changes in fabric properties and structure. The incorporation of absorbable yarns into knitted fabrics did not result in the same level change in fabric structure and property as woven fabrics. In both W and K groups, a minimal level of mechanical strength of the fabrics was maintained due to the remaining Dacron yarns. Structural integrity of these fabrics was retained at the end of hydrolytic degradation study. The data obtained could be used to correlate with the subsequent in vivo performance of these bicomponent vascular grafts. If correlations exist, they could be used to improve the design of future bicomponent vascular grafts for improved performance. PMID- 8245048 TI - Degradation of polyurethane foams used in the Meme breast implant. AB - The release of toluene diisocyanate derivatives from polyurethane foam covers that were removed from the Meme breast implant was studied. The polyurethane foam covers as retrieved from the implants were contaminated on average with 2,4- and 2,6-toluene diamine (TDA), 2,4- and 2,6-toluene diisocyanate (TDI), and toluene isocyanate amine (TIA) at the 1086 ppm level (combined TDA, TDI, and TIA). When cleaned foam was incubated in Ringer's solution at 37 degrees C for periods ranging from 6-35 days, 4.6 ppm/day of TDA, TDI, and TIA (combined) was formed; the projected annual degradation rate of the foam under these conditions was estimated to be about 0.8%. When cleaned foam was incubated dry at 37 degrees C, 1.5 ppm/day of TDA, TDI, and TIA (combined) was produced. Hence, the polyurethane foam covers used in the Meme breast implants are susceptible to both thermal and hydrolytic degradation under simulated physiological conditions. PMID- 8245049 TI - Outcomes measurements. PMID- 8245050 TI - A self-administered questionnaire for the assessment of severity of symptoms and functional status in carpal tunnel syndrome. AB - We developed a self-administered questionnaire for the assessment of severity of symptoms and functional status in patients who have carpal tunnel syndrome. The reproducibility, internal consistency, validity, and responsiveness to clinical change of scales for the measurement of severity of symptoms and functional status were evaluated in a clinical study. The scales were highly reproducible (Pearson correlation coefficient, r = 0.91 and 0.93 for severity of symptoms and functional status, respectively) and internally consistent (Cronbach alpha, 0.89 and 0.91 for severity of symptoms and functional status, respectively). Both scales had positive, but modest or weak, correlations with two-point discrimination and Semmes-Weinstein monofilament testing (Spearman coefficient, r = 0.12 to 0.42). In thirty-eight patients who were operated on in 1990 and were evaluated a median of fourteen months postoperatively, the mean symptom-severity score improved from 3.4 points preoperatively to 1.9 points at the latest follow up examination, while the mean functional-status score improved from 3 to 2 points (5 points is the worst score and 1 point is the best score for each scale). Similar improvement was noted in twenty-six patients who were evaluated before and three months after the operation. We concluded that the scales for the measurement of severity of symptoms and functional status are reproducible, internally consistent, and responsive to clinical change, and that they measure dimensions of outcomes not captured by traditional measurements of impairment of the median nerve. These scales should enhance standardization of measurement of outcomes in studies of treatment for carpal tunnel syndrome. PMID- 8245051 TI - Late-onset pseudarthrosis of the dysplastic tibia. AB - Eleven children in whom a tibial fracture occurred after minor trauma had pre existing dysplastic changes evident radiographically. These changes included cortical tapering, sclerosis, and formation of a cyst in the region of the medullary canal. Ten of the eleven patients had had no more fractures an average of fifteen years after the most recent fracture. Six of the fractures healed following prolonged immobilization in a cast, but four of the six tibiae were abnormally bowed anteriorly, and it was thought that a stress fracture could occur in the future. Four of the patients had a clinically straight tibia and radiographic evidence of thick cortices following corrective osteotomy, intramedullary fixation with bone-grafting, and prolonged immobilization in a cast. At the most recent follow-up examination, the eleventh patient had a persistent pseudarthrosis despite several operative procedures. While there were too few patients in this series for us to draw definite conclusions, our findings suggest that late-onset pseudarthrosis of a dysplastic tibia has a better prognosis than does congenital pseudarthrosis. PMID- 8245052 TI - Use of the metaphyseal-diaphyseal angle in the evaluation of bowed legs. AB - We evaluated the accuracy of the angle described by Levine and Drennan, the metaphyseal-diaphyseal angle of the proximal aspect of the tibia, for the differentiation of physiological bowing from Blount disease. We compared this angle, as measured at presentation, in 106 children (179 extremities) who had physiological bowing with the angle in nineteen children (thirty-two extremities) who had documented Blount disease. The angle averaged 9 +/- 3.9 degrees for the patients who had physiological bowing and 19 +/- 5.7 degrees for the patients who had Blount disease (p < 0.0000001). Linear regression analysis, performed to evaluate any changes in the metaphyseal-diaphyseal angle in relation to age, showed that the older the child was at the time of presentation the more likely it was that the angle would be smaller in a child who had physiological bowing and larger in a child who had Blount disease. The chance for false-positive and false-negative errors was greater than 5 per cent if the angle was more than 9 degrees but less than 16 degrees. Sixty-six (37 percent) of the 179 extremities in the group that had physiological bowing had an angle of at least 11 degrees; one extremity affected by Blount disease had an angle of less than 11 degrees. In this study, we found that the metaphyseal-diaphyseal angle may be helpful in the identification of Blount disease but should not be the sole criterion used to determine the diagnosis. PMID- 8245053 TI - The Mitchell procedure for the treatment of adolescent hallux valgus. A long-term study. AB - Thirty patients who had had a total of fifty-one Mitchell procedures to correct adolescent hallux valgus deformities were examined clinically and radiographically an average of seven years (range, three to seventeen years) after the operation. The average age of the patients at the time of the operation was fifteen years (range, ten to nineteen years). The result was excellent in nineteen feet, good in sixteen, fair in six, and poor in ten. The fair and poor results were associated with recurrence of the deformity, stiffness (real or perceived), and unsightly scars. The cause of the fair and poor results was inadequate correction at the time of the operation in six feet and loss of fixation in two; the loss of fixation resulted in one recurrence and one malunion. The remaining eight patients who had a fair or poor result were not totally satisfied and had reservations about more than one of three categories (relief of discomfort, appearance, or range of motion). Seventeen feet had a plantar callosity beneath the second metatarsal head, suggesting increased load bearing by the second metatarsal. Although sixteen callosities caused no symptoms at the most recent follow-up evaluation, the long-term implications of this altered pattern of weight-bearing are unknown. PMID- 8245054 TI - The effect of elective total hip replacement on health-related quality of life. AB - The effect of total hip replacement on the health-related quality of life of patients who have osteoarthrosis was examined as part of a randomized, controlled trial comparing femoral head prostheses that were inserted with or without cement. One hundred and eighty-eight patients were followed for three months: 179 of them, for six months; 156, for one year; and ninety, for two years. The health related quality of life was assessed with use of the Harris hip score, the Merle d'Aubigne hip score, the Sickness Impact Profile, the Western Ontario and McMaster University (WOMAC) Osteoarthritis Index, the McMaster--Toronto Arthritis (MACTAR) Patient Preference Disability Questionnaire, and the time trade-off technique as a measure of utility. Patients also took the six-minute-walk test. The mean age of the patients in the study was sixty-four years (range, forty to seventy-five years); ninety-seven patients (53 per cent) were men and ninety-four (50 per cent) had a prosthesis inserted with cement. Only three of 188 patients refused to return for quality-of-life assessments. There was significant improvement in all health-related quality-of-life measures and in the six-minute walk test after the operation (p < 0.01 for all items, except for the work dimension of the Sickness Impact Profile at three months [p = 0.07]). Most of the improvement had occurred by three months postoperatively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245055 TI - Severe osteolysis of the pelvic in association with acetabular replacement without cement. AB - We reviewed the cases of fourteen patients (fifteen lesions) who had osteolysis following the replacement of the acetabulum without cement. Nine women and five men, seventeen to sixty-seven years old, were involved in the study. One woman had bilateral pelvic osteolysis. Eight of the fifteen index acetabular reconstructions were done with a titanium-alloy implant and seven, with a chromium-cobalt-alloy implant. Eleven of the fifteen acetabular components had holes in the metal shell that may have acted as a conduit through which wear debris could gain access to the implant-bone interface, but only two of the acetabular components had been fixed with screws. In these two acetabular components, all available screw holes were not filled. The polyethylene liner was eight millimeters thick or less in twelve of the fifteen acetabular components; all of the liners were ten millimeters thick or less. The diameter of the head of eleven of the fifteen femoral components was thirty-two millimeters. Fourteen of the fifteen femoral components were placed without cement, and all but one was radiographically stable. The duration from the index operation to the appearance of pelvic osteolysis ranged from fifty-three to eighty-four months (mean, sixty five months). At the time of the diagnosis, the patients were functioning well clinically, and all but three had a Harris hip score of 90 points or better, despite extensive destruction of bone in some instances. Since these patients were functioning well, the pelvic osteolysis was diagnosed radiographically at a regular follow-up examination. Only one patient had evidence of migration of the acetabular component on serial radiographs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245056 TI - Passive motion: the dose effects on joint stiffness, muscle mass, bone density, and regional swelling. A study in an experimental model following intra-articular injury. AB - We studied the effects of passive motion on joint stiffness, muscle mass, bone density, and regional swelling after an intra-articular injury. Instrumentation was applied to the hindlimbs of thirty adolescent New Zealand White rabbits to allow either passive motion or immobilization of the ankle. The knee was immobilized by the locking together of Steinmann pins that had been placed within the medullary canals of the tibia and femur. An intra-articular injury was produced by drilling of the tibial pin through the ankle joint into the talus and subsequent withdrawal of the pin from the ankle joint. The rabbits were divided into five groups, and they received four, eight, twelve, sixteen, or twenty-four hours of passive motion each day during the three-week period of study. One ankle of each rabbit was moved through an arc of 90 to 170 degrees of dorsiflexion at one cycle per minute, while the contralateral ankle was immobilized in 100 degrees of dorsiflexion with an aluminum splint, which was fixed to the aluminum block that was used to stabilize the knee joint. We found that sixteen and twenty four hours of passive motion prevented stiffness of the joint. Passive motion for shorter periods was ineffective, even harmful, and resulted in stiffness ratios that were as much as four times higher than those of the control limbs (those treated with immobilization). Swelling of the limb decreased only in the group that received twenty-four hours of passive motion. Muscle mass increased by an average of 13 per cent (range, 4 to 34 per cent), in comparison with that of the immobilized limbs in every group that was treated with passive motion. Bone density was maintained only in the limbs in which the ankle became stiff (ankles that had been treated with passive motion for twelve hours or less). An inverse relationship was noted between the duration of passive motion and the radiographic density of the distal tibial metaphysis; this relationship was statistically significant (p < 0.01). The limbs treated for twelve, eight, or four hours each day showed progressively greater bone density in comparison with those treated with immobilization or with sixteen or twenty-four hours of passive motion. PMID- 8245057 TI - Intralesional excision compared with en bloc resection for giant-cell tumors of bone. AB - We reviewed the results for forty consecutively seen patients, each of whom had been managed by one surgeon for a giant-cell tumor of bone in an extremity between 1976 and 1990. Twenty patients had been managed with an en bloc resection and twenty, with an intralesional excision of the tumor with adjunctive local insertion of methylmethacrylate or phenol. All patients had been followed for a minimum of two years. Both en bloc resection and intralesional excision were found to be excellent oncological procedures. There were fewer complications and better functional results after the intralesional procedure than following the en bloc resection. PMID- 8245058 TI - The use of a bone allograft for reconstruction after resection of giant-cell tumor close to the knee. AB - The results of bone-allograft reconstruction after the resection of giant-cell tumor close to the knee were reviewed in fifty-two patients (fifty-five allografts), who had been followed for a mean of seven years (range, two to twenty-four years). One giant-cell tumor was graded as stage 1; twenty, as stage 2; and thirty-one, as stage 3. Three reconstructions were repeated transplants that were done after the failure of a previous transplant. Ten allograft reconstructions were intercalary and were combined with an arthrodesis of the knee, and forty-five were osteoarticular. Major complications included infection (after three reconstructions), resorption of the graft (six), collapse of the articular surface (two), fracture (two), and recurrence (one). According to the criteria described by Mankin et al. for functional analysis, forty-two (76 per cent) of the extremities had a result that was considered to be excellent or good. Radiographic evaluation according to the system of the Musculoskeletal Tumor Society showed a mean score of 72 per cent for osteoarticular reconstructions, and of 86 per cent for intercalary reconstructions. PMID- 8245059 TI - Fractures in large-segment allografts. AB - Sixteen of thirty-five large-segment allografts that had been implanted after resection for neoplastic conditions, and had been followed for a minimum of thirty-six months, were found to have fractured at a mean of twenty-six months after the implantation. Thirteen of the fractures were treated operatively, and we found a lack of vascularization and soft-tissue attachments to the graft at the fracture site. For seven fractured grafts, there were radiographic and clinical signs of union with the host bone. Eight of the sixteen grafts that had fractured were salvaged with one or more autogenous bone grafts, and two healed spontaneously. Thus, twenty-nine of the thirty-five grafting procedures were considered to have been successful in that the initial objective--provision of a functional segment for skeletal replacement--had been achieved. Multivariate analysis revealed a significant correlation for fracture in patients who were receiving chemotherapy when internal fixation of the graft had included devices that penetrated the cortices of the graft (p < 0.05). PMID- 8245060 TI - A biomechanical analysis of decompression and reconstruction methods in the cervical spine. Emphasis on a carbon-fiber-composite cage. AB - Biomechanical analysis of three different patterns of instability--that created by fifth and sixth cervical anterior discectomy, that created by one-level (fifth cervical) anterior corpectomy, and that created by two-level (fourth and fifth cervical) corpectomy--was performed in eighteen calf spines. Three types of anterior reconstruction--anterior iliac strut bone-grafting, use of an anterior carbon-fiber-composite cage packed with cancellous bone graft, as well as use of polymethylmethacrylate anteriorly--were cyclically tested in axial compression, torsion, and flexion-extension. Each of these types of reconstruction was also tested with supplemental posterior wire stabilization (the triple-wire technique of Bohlman). Regardless of the type of anterior instability, the carbon-fiber reinforced cage packed with cancellous bone graft was more rigid than the iliac bone graft alone. The cage resulted in good stiffness in the axial compression and rotation tests and was the most rigid construct in the flexion-extension tests. The superior aspect of the polymethylmethacrylate constructs loosened at the bone-cement interface in eight of the twelve specimens during flexion extension testing. The addition of the supplemental posterior wiring to the anterior constructs provided additional rigidity in flexion-extension testing. PMID- 8245061 TI - Coxsackie-B monarthritis with hepatitis. A case report. PMID- 8245062 TI - Tuberculous peroneal tenosynovitis. A case report. PMID- 8245063 TI - Late effect of isolated avulsion fractures of the lesser tubercle of the humerus in children. Report of two cases. PMID- 8245064 TI - Traumatic lumbopelvic spondyloptosis. A case report. PMID- 8245065 TI - The diagnosis and orthopaedic treatment of childhood spinal muscular atrophy, peripheral neuropathy, Friedreich ataxia, and arthrogryposis. PMID- 8245066 TI - Posterior stabilized prosthesis. Results after follow-up of nine to twelve years. PMID- 8245067 TI - The value of aspiration of the hip joint before revision total hip arthroplasty. PMID- 8245068 TI - Biological effects of intramedullary reaming. PMID- 8245069 TI - Metal backing of polyethylene implants. PMID- 8245070 TI - Fractures caused by child abuse. PMID- 8245071 TI - Triple innominate osteotomy for hip stabilisation and transiliac leg lengthening after poliomyelitis. AB - We reviewed our experience with a modified triple innominate osteotomy for hip instability and limb shortening due to poliomyelitis in 62 adolescent and adult patients, treated from 1973 to 1990. Their ages at surgery ranged from 12 years to 35 years (average 22.3). At a mean follow-up of 4 years (2 to 18) 59 of the patients (95.2%) had substantial improvement in hip stability, and all but one had radiological improvement as determined by the acetabular angle, centre-edge angle and acetabulum-head quotient. In 59 cases in which transiliac limb lengthening was attempted, the mean gain was 1.7 cm (0.6 to 3.0). When the abductor muscles had been partially paralysed, the operation produced an appreciable increase in power in 12 of the 39 hips examined. PMID- 8245072 TI - Loosening and bone resorption in exeter hip arthroplasties. Review at a minimum of five years. AB - We reviewed at a minimum elapsed time of five years a consecutive series of 143 primary Exeter hip replacements in which matt-surfaced femoral stems had been used. Twenty-five patients had died and six stems and two sockets had been revised before follow-up. The remaining 110 hips were all examined clinically and radiographically. In 15 hips there were radiographic signs of definite loosening of the stem and in eight suspected loosening. The acetabulum was loose in four hips. In another eight hips localised bone resorption was present without signs of loosening. Half the patients with loosening or localised bone resorption had mild pain or no pain at all. The late complication rate with the matt-surfaced Exeter femoral stem is unacceptably high. PMID- 8245073 TI - Cement-within-cement revision hip arthroplasty. AB - We reviewed 19 revision hip arthroplasties in which the new femoral component had been recemented into the old, intact cement mantle. The mean time from the first operation to revision was 64 months and the average follow-up was 59 months. There were 7 excellent, 11 good, and one fair result. No femoral component had been revised for loosening and all the stems appeared radiographically stable. Complications included intraoperative perforation of the femur on two occasions and one dislocation. The use of the cement-within-cement technique requires that the old cement surface be dry and roughened to increase the surface area and that the cement be injected in the liquid phase to prevent lamination. The indications for this technique include a broken stem with an intact distal cement mantle, the removal of a femoral component for revision of a loose cup to improve exposure and/or increase offset, recurrent dislocation secondary to component malposition, and debonding of the femoral component within an intact cement mantle. PMID- 8245074 TI - Results of cemented total hip replacement in juvenile chronic arthritis. A radiological review. AB - We assessed 57 total hip arthroplasties in 34 adolescents with juvenile chronic arthritis using standard radiological techniques at an average of 4.7 years (20 months to 9 years) after surgery. The incidence of overall loosening was 24.6% (14 hips), but hips with a follow-up of more than five years had a loosening rate of 43.5% (10 hips; p < 0.01). PMID- 8245075 TI - Avascular necrosis of the femoral head in sickle-cell disease. Treatment of collapse by the injection of acrylic cement. AB - In ten patients with sickle-cell disease, we used a new technique of cement injection for the treatment of 16 painful hips with a radiographic crescent line or flattening of the articular surface due to avascular necrosis. The necrotic bone and overlying cartilage are elevated by the injection to restore the sphericity of the femoral head. Five days after the operation, full weight bearing was allowed with the help of crutches for three weeks. The time in hospital averaged eight days; the average blood loss was 100 ml. There was early pain relief and postoperative radiographs showed improvement in the shape of the femoral head. At a mean follow-up of 5 years (3 to 7), 14 of the 16 hips were still improved although some gave slight pain. Only two hips had required revision to total hip arthroplasty, at one year and two years respectively. The increasing longevity of patients with sickle-cell disease means that avascular necrosis will be an increasing problem. Total hip replacement has a poor prognosis because of the risks of infection, high blood loss, and early loosening. Cement injection does not have these problems and allows for earlier, more conservative surgery. PMID- 8245076 TI - Costoplasty in adolescent idiopathic scoliosis. Objective results in 55 patients. AB - Costoplasty can reduce the important cosmetic deformity of rib prominence in scoliosis but there are few objective reports of correction. We recorded the results of three objective methods of assessing back shape before and after short segment costoplasty in 55 patients. We showed that significant improvement was maintained over a two-year follow-up period. Primary costoplasty at the time of scoliosis surgery (n = 35) achieved greater proportional correction than secondary costoplasty performed after fusion of the spine (n = 20). The rib segments removed at primary surgery provided enough bone for the autogenous graft; harvesting from the pelvis was unnecessary. We report a new classification of rib morphology which helps in planning the site and extent of costoplasty, and in predicting the possible correction. PMID- 8245077 TI - Extracoelomic approach to the spine. AB - A modified transthoracic approach to the thoracic vertebral column is described. In this method, the parietal pleura is detached from the chest wall and retracted with the visceral pleura and its contents. A direct approach to the vertebral bodies is thus achieved without transgression of the intrapleural space. The technique can be extended to include exposure of the thoracolumbar spinal column, utilising a thoracoabdominal approach with extrapleural and extraperitoneal dissection. Management of the costophrenic detachment is thus simplified. This approach has significant advantages for orthopaedic, vascular and neurosurgical procedures. PMID- 8245078 TI - Repeat decompression of lumbar nerve roots. A prospective two-year evaluation. AB - In a prospective, consecutive study 93 patients who had had previous lumbar spinal surgery underwent repeat decompression for persistent or recurrent back and leg pain. The previous operations had been discectomies in 65 patients and decompression for spinal stenosis in 28; two of the latter group had also had posterolateral fusion. At the repeat operation, disc herniation was found in 19 patients, lateral spinal stenosis in 19, central spinal stenosis in 20 and periradicular fibrosis in 35. Ninety-one patients were followed up for two years after surgery; the effect of the operation was recorded using a four-scale grading system. The results were significantly related to the diagnosis. Nerve root compression due to recurrent disc herniation or to bony compression responded well to repeat decompression. In patients with a single nerve-root compression the results were similar to those obtained in primary operations. Sciatica due to nerve-root scarring was seldom improved by the repeat operation. PMID- 8245079 TI - Longitudinal growth of the rabbit tibia after callotasis. AB - We investigated the effect of bone lengthening by callotasis on longitudinal growth of the tibia in rabbits. Ninety-nine five-week-old immature rabbits were divided into five groups according to the percentage of lengthening: group I, 10%; group II, 20%; group III, 30%; group IV, 40%; and group V, sham operation without lengthening. Corticotomy was performed at the proximal metaphysis of the left tibia and the right tibia was used as a control. The lengthening rate was 0.25 mm twice daily. Radiological, histomorphometric, and immunohistochemical studies were done on animals killed at the time of corticotomy, at the completion of lengthening, and thereafter every two weeks until 12 weeks. Tibial lengthening did not cause retardation of growth when the bone was lengthened by up to 20%. When the bone was lengthened by 30% or more, growth retardation was evident, and persisted until skeletal maturity. PMID- 8245080 TI - Massive allografts sterilised by irradiation. Clinical results. AB - From 1984 to 1988 we implanted 127 massive allografts irradiated with a dose of 25,000 grays. These were reviewed at a minimum follow-up of three years to determine the effect of irradiation on infection, the complications and the functional result. No bacteriological infection was seen in the 44 patients who had allografts for revision of joint arthroplasty or for a tumour with no adjuvant therapy. For the 83 patients who also had chemotherapy or radiotherapy or both for a bone tumour, the rate of infection was 13%. The major mechanical complications were nonunion in seven grafts (5.5%) and fracture in eight (6%). These rates do not differ greatly from those reported for non-irradiated grafts. Our results suggest that irradiation, which remains the most convenient and acceptable method of sterilisation, does not jeopardize the clinical results. PMID- 8245081 TI - Bone density after rigid plate fixation of tibial fractures. A dual-energy X-ray absorptiometry study. AB - We used dual-energy X-ray absorptiometry (DEXA) to measure the bone mineral content (BMC) of both tibiae in 13 patients who had been treated for a tibial fracture by rigid plate fixation. Within two weeks of plate removal the BMC was significantly greater in the bone that had been under the plate than at the same site in the control tibia. An unplated area of bone near the ankle showed a significant decrease in BMC at the time of plate removal with subsequent return to the level of the control tibia during the ensuing 18 months. We conclude that osteoinductive influences outweigh the potential causes of osteopenia, such as stress shielding and disuse, and that, contrary to expectation, demineralisation is not a factor in the diminished strength of the tibia after plating for fracture. PMID- 8245082 TI - Glove perforation during hip arthroplasty. A randomised prospective study of a new taperpoint needle. AB - Exposure to blood is a hazard for all surgeons. We assessed the incidence of glove perforation and needlestick injury from a new blunt taperpoint needle designed to penetrate tissues other than skin with the minimum of force. We performed a prospective, randomised trial comparing the incidence of perforations of surgical gloves with the new needle and a standard cutting needle during wound closure after hip arthroplasties. There was at least one glove perforation in 46 of 69 such procedures (67%). The use of the taperpoint needle produced a significant decrease in perforations (p = 0.049). PMID- 8245083 TI - The detection of fat embolism by transoesophageal echocardiography during reamed intramedullary nailing. A study of 24 patients with femoral and tibial fractures. AB - We performed transoesophageal echocardiography on 24 patients during reamed intramedullary nailing of 17 tibial and seven femoral fractures. In 14 patients there was only minimal evidence of emboli passing through the heart, but in six copious showers of small emboli (< 10 mm maximum dimension) were observed. In four other patients, there were also multiple large emboli (> 10 mm maximum dimension). Three of these patients developed fat embolism syndrome postoperatively and one died. Earlier nailing was associated with smaller quantities of emboli. PMID- 8245084 TI - Central grafting for persistent nonunion of the tibia. A lateral approach to the tibia, creating a central compartment. AB - We report the operative technique and results of a new method of central grafting for persistent nonunion of the tibial shaft. The operation is performed through a lateral approach, anterior to the fibula. Fresh autogenous bone from the iliac crest is used to form a central bridge between the tibia and fibula above, below and at the level of the nonunion. In 48 tibiae, most with long-standing nonunion and some with infection or bone defects, sound healing was obtained in 45 after one operation. Only one failure needed amputation. PMID- 8245085 TI - Intra-articular fractures of the calcaneum treated operatively or conservatively. A prospective study. AB - We report a prospective trial of 66 patients with intraarticular fractures of the calcaneum. All fractures were assessed by CT. Patients with displaced fractures were randomised to receive either conservative (n = 31) or operative treatment (n = 25). Undisplaced fractures (n = 10) were treated conservatively. Operation involved open reduction of the posterior subtalar joint, and fixation with Kirschner wires. All 66 patients were reviewed at a minimum of one year (mean 23 months). After conservative treatment the undisplaced fractures had slightly better results than the displaced fractures. There was no significant difference in outcome between the operatively and the conservatively treated displaced fractures. We have also documented prospectively the natural history of the injury, which is of use in assessing prognosis for both clinical and medicolegal purposes. PMID- 8245086 TI - Repair of avulsed insertion of biceps. A new technique in four cases. AB - We report the use of the Mitek anchor with a limited approach for repair of avulsion of the biceps tendon from the radius in four middle-aged men. All regained a full range of movement with minimal loss of power. PMID- 8245087 TI - The incidence of fatal pulmonary embolism after knee replacement with no prophylactic anticoagulation. AB - We made a prospective study of the incidence of fatal pulmonary embolism in patients after total knee replacement with no prophylactic anticoagulation. There were 499 consecutive patients having 527 knee replacements. They all wore anti thromboembolic stockings and were mobilised 48 hours after surgery. No patient was lost to follow-up. One patient died of pulmonary embolism 22 days after operation. There were no other deaths within three months of operation. The incidence of fatal pulmonary embolism was 0.19% (95% confidence interval: 0 to 0.6%). Fatal pulmonary embolism is rare after total knee replacement without prophylactic anticoagulation and the routine anticoagulation of these patients is of doubtful value. PMID- 8245088 TI - The incidence of deep-vein thrombosis after upper tibial osteotomy. A venographic study. AB - We performed postoperative venography on 84 consecutive patients who had undergone upper tibial osteotomy for medial compartment osteoarthritis of the knee. Deep-vein thrombosis was demonstrated in 41%. Only 15% of the cases were diagnosed clinically, all in the calf veins. Cases of proximal thromboses (3) and mixed-vein thromboses (12) were only revealed by venography. PMID- 8245089 TI - Acute haemarthrosis of the knee in athletes. A prospective study of 106 cases. AB - We made a prospective arthroscopic study of 106 skeletally mature male sportsmen with an average age of 28.35 years (16.8 to 44) who presented with an acute haemarthrosis of the knee due to sporting activities. We excluded those with patellar dislocations, radiographic bone injuries, extra-articular ligamentous lesions or a previous injury to the same joint. The anterior cruciate ligament (ACL) was intact in 35 patients, partially disrupted in 28 and completely ruptured in 43. In the patients with an ACL lesion, associated injuries included meniscal tears (17 patients), cartilaginous loose bodies (6), and minimal osteochondral fractures of the patella (2), the tibial plateau (3) or the femoral condyle (9). We found no age-related trend in the pattern of ACL injuries. Isolated injuries included one small osteochondral fracture of the patella, and one partial and one total disruption of the posterior cruciate ligament. Three patients had cartilaginous loose bodies, and no injury was detected in five. Acute traumatic haemarthrosis indicates a serious ligament injury until proved otherwise, and arthroscopy is needed to complement careful history and clinical examination. All cases with a tense effusion developing within 12 hours of injury should have an aspiration. If haemarthrosis is confirmed, urgent admission and arthroscopy are indicated. PMID- 8245090 TI - Factors affecting the range of movement of total knee arthroplasty. AB - We have investigated those factors which influence the range of movement after total knee arthroplasty, including sex, age, preoperative diagnosis and preoperative flexion deformity and flexion range. We also compared cemented and cementless tibial fixation, the influence of collateral ligament and lateral parapatellar releases and of replacement of the patella, and of the period of postoperative immobilisation. We reviewed 516 Johnson-Elloy (Accord) knee arthroplasties performed between 1982 and 1989, with a minimum follow-up of 12 months. The most important factors in the range of flexion achieved after arthroplasty are the diagnosis and the preoperative range of flexion. In patients with osteoarthritis there was a mean loss of flexion; in rheumatoid arthritis there was a mean gain. In both groups, the stiffer knees gained motion and the more mobile knees lost it. Post-operative range of motion was not influenced significantly by cement fixation, collateral ligament or patellar retinacular releases, prolonged immobilisation or patellar replacement. PMID- 8245091 TI - Chondrolysis in slipped upper femoral epiphysis. Long-term study of the aetiology and natural history. AB - Of 44 patients (55 hips) with slipped upper femoral epiphysis treated from 1963 to 1989, 13 (14 hips) developed chondrolysis. Eight hips had chondrolysis at the time of presentation, all in female patients who were either coloured or black and who had moderate or severe slips. The other six hips had persistent pin penetration of the joint; in five of these the pin penetrated the anterosuperior quadrant of the head. Removal of penetrating pins resulted in improvement in pain in all six hips and in the range of movement in four. Chondrolysis did not develop in any of 11 hips with transient intraoperative pin penetration. In hips with chondrolysis maximum joint-space narrowing developed within the first year; improvement in joint space and range of movement continued for up to three years after maximal involvement. At an average follow-up of 13.3 years no patient had pain but five hips were stiff. PMID- 8245092 TI - Double oblique diaphyseal osteotomy. A new technique for lengthening deformed and short lower limbs. AB - We treated 25 patients with severely deformed and shortened lower limbs by double oblique diaphyseal osteotomy of the short deformed bone, followed by balanced skeletal traction to correct the alignment and increase the length of the limb. The technique is simple and inexpensive and does not require special equipment. It has the advantage of allowing rapid lengthening and secure healing as the overlap allows end-to-end union with no bone gap. The lengthening achieved ranged from 6 to 16 cm. No serious complications were encountered. PMID- 8245093 TI - Medial calcaneal osteotomy for relapsed equinovarus deformity. Long-term study of the results of Frederick Dwyer. AB - In the 1950s Frederick Dwyer evolved the concept of treating resistant and relapsed clubfoot by osteotomy of the calcaneum. He published the results of his medial opening wedge procedure in 1963 with a mean follow-up of five years. We present the structured, radiographic and functional results at a mean elapsed time of 27 years of 36 feet (26 patients) all operated on by Dwyer. Their mean Laaveg and Ponseti (1980) grading was 83.7%. In 94% the heel was in neutral or valgus and 86% of the feet were plantigrade. A good range of movement was present in the ankle and subtalar joints in 83%. PMID- 8245094 TI - Cardiac arrest during intramedullary nailing for femoral metastases. PMID- 8245095 TI - Scaphoid fracture at the tip of a Herbert screw. PMID- 8245096 TI - The effect of complications after hip fracture on rehabilitation. PMID- 8245097 TI - Recurrence of an osteosarcoma in a needle biopsy track. PMID- 8245098 TI - Trochanteric osteotomy. PMID- 8245099 TI - Intra-articular fractures of the calcaneum. PMID- 8245100 TI - Wear particles and lymphadenopathy. PMID- 8245101 TI - Measurement of polyethylene wear. PMID- 8245102 TI - Composite grafts of human keratinocytes grown on a polyglactin mesh-cultured fibroblast dermal substitute function as a bilayer skin replacement in full thickness wounds on athymic mice. AB - We have developed and tested in athymic mice a new, cultured, dermal-epidermal graft composed of two human cell types coupled with a biodegradable dermal scaffold. Cultured, proliferating human keratinocytes (HK) were applied to the surface of a living dermal tissue replacement that is composed of human fibroblasts cultured on a polyglactin mesh. After 4 to 6 days of coculture, proliferating HKs achieved confluency on the surface of the living dermal tissue replacement. Grafts were then transferred to full-thickness wounds on the dorsum of athymic mice. Sixteen animals were grafted, and the mean percentage of graft take (original wound area covered) on day 20 after grafting was 51.25%. Staining with antibody specific for human involucrin confirmed the presence of HKs on closed wounds, and staining with antibody specific for human laminin revealed a continuous layer of laminin at the dermal-epidermal junction on day 20. Animals closed with living dermal tissue replacement alone markedly contracted, whereas application of living dermal tissue replacement-HK grafts appeared to retard contraction. Because polyglactin mesh fibers are absorbed by hydrolysis rather than by enzymatic degradation, this living composite graft may be more resistant to destruction when placed on excised human wounds than are composite grafts, which are composed of a collagen matrix. The inclusion of the living dermal substitute may ultimately provide better skin quality than is achieved from the use of cultured keratinocytes alone. Fragility of the epidermal layer is probably due to the short-term culture of HKs on the living dermal tissue replacement, and further efforts to develop a thicker epithelial layer may improve graft durability. PMID- 8245103 TI - Effects of fluid resuscitation, burn eschar excision, and blockade of afferent pain responses on bacterial translocation and acid-base balance after murine burn injury. AB - We tested effects of fluid resuscitation, early burn excision/grafting, and blockade of afferent stimuli from the burn wound on bacterial translocation and acid-base balance after murine burn injury. Burn excisions were performed with patients either 15 minutes or 2 hours after burn injury under anesthesia, and excised wounds were immediately closed with murine allograft skin. Twenty-four hours after 25% total body surface area (TBSA) burn injury and 48 hours after 32% TBSA injury, mesenteric lymph nodes were cultured. Incidences of bacterial translocation in 25% and 32% TBSA burns were 31.6% and 68.4% of animals, respectively. Burned animals were in severe shock, and metabolic acidosis reached a nadir 12 hours after burn injury, with base deficit -27.8 +/- 0.6 mEq/L; 5% to 10% of animals died acutely after burn injury. After excision/grafting of burned mice at 2 hours after burn injury, the incidence of bacterial translocation was unchanged (35.7% with 25% TBSA burn, 73.3% with 32% TBSA burn), and mortality did not change. When 32% TBSA excisions were performed exactly 10 minutes after burn injury, four of the 13 mice died within several hours, and five (55.5%) of the nine survivors translocated. Rates of bacterial translocation in mice receiving anesthesia or excision/grafting without burn injury were 15.0% and 20%, respectively (p = NS compared with normal mice). Subcutaneous implantation of normal or burned skin into normal animals neither elicited shock nor increased the incidence of bacterial translocation. Increasing amounts of fluid resuscitation in the 25% TBSA burn model provided only delayed improvement of acid-base balance; increased amounts of fluid did not decrease bacterial translocation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245104 TI - Success of immediate intragastric feeding of patients with burns. AB - We evaluated our attempts to aggressively feed 45 adult patients with burns with the use of intragastric tube feedings. Patients were divided into three burn sizes (20% to 30% total body surface area, 31% to 44% total body surface area, and > 44% total body surface area). No attempts were made to place feeding tubes across the pylorus. Patients were fed as early as possible after admission. Mean caloric intake met calculated nutritional goals beginning on the second day after burn injury, and in the severely burned group, mean caloric intake was 2500+ kcal by the second day. Initiation of intragastric feedings within hours of burn injury results in the provision of high daily levels of nutrition and may obviate the necessity of placing a transpyloric feeding tube in the majority of patients with burns. PMID- 8245105 TI - Intragastric pH monitoring. AB - Buffering of intragastric pH is an accepted treatment modality for prophylaxis against the development of gastric stress ulcers. This method of prophylaxis is commonly based on the pH value acquired by measurement of gastric aspirate. Recent literature suggests pH measurement techniques that involve gastric aspirate specimens have many flaws. The purpose of this study was to compare gastric pH measurements with the use of a nasogastric sensor, meter system, and pH-sensitive test paper. Fifteen hundred paired serial measurements of intragastric pH were obtained on 19 thermally injured patients (16 men and three women, ages 23 to 79 years, total body surface area burn 25% to 80%). A double lumen tube containing an antimony/graphite pH sensor incorporated into the tip of the tube was inserted with the use of a standard technique. Each tube was in place an average of 5.7 days (range 1 to 15 days). Patients were randomized into two groups. The first group (six patients) received non-acid-buffering prophylaxis therapy. The second group (13 patients) received standard antacid or antacid/H2 histamine-blocking agent combination prophylaxis therapy. Analysis of the 539 paired measurements for the non-acid-buffering revealed a correlation coefficient of r = 0.532. The 961 measurements from the group receiving gastric acid buffering revealed a correlation coefficient of r = 0.569. Paired t test values for the sample showed a significant difference (18.52, p < 0.0000) between measurement techniques. PMID- 8245106 TI - Management of burns of the penis. AB - Burns of the penis can be physically and psychologically devastating. This is particularly true in young pediatric patients who have to content with growth as well as with the trials of adolescence and the development of self-image. Little has been written on the grafting management of these injuries. This report reviews a management technique used with great success. Ten patients who sustained full-thickness penis burns were reviewed. The grafting technique consisted of excision and grafting with sheet grafts in a one- or two-stage setting. After grafting, the penis was placed in traction with a weighted Foley catheter. Grafts were left open or stented. No complications occurred from the traction apparatus, and graft take was uniformly excellent. Short-term results have been excellent, and no urethral problems have developed over a median 3-year follow-up. This technique appears to provide excellent means of supporting the penis to accomplish superb graft take in this hard-to-manage area. PMID- 8245107 TI - Transient and stable ionic permeabilization of isolated skeletal muscle cells after electrical shock. AB - Electroporation of skeletal muscle cell membranes has been postulated to cause myonecrosis in victims of major electrical trauma. To evaluate this concept we carried out a series of experiments to measure the transmembrane potential (delta Vm) threshold for skeletal muscle membrane electroporation using isolated mammalian skeletal muscle cells and compared this threshold with the expected range of delta Vm in victims of electrical trauma. Alterations in membrane Mg2+ or Ca2+ permeability in response to applied extracellular field pulses were quantified by measuring the kinetics of influx before and after field exposure. To avoid heating effects, 4-msec duration field pulses were used. The resting intracellular [Mg2+] was 0.86 +/- 0.01 mmol/l, and [Ca2+] was 0.1 +/- 0.01 microns. The delta Vm threshold for transient or stable electropore formation was determined by performing experiments over a wide range of applied fields. A delta Vm of 340 mV produced no significant change in intracellular [Mg2+]. Delta Vms ranging between 340 to 480 mV caused only a transient influx of Mg2+, indicating that spontaneous sealing of the membrane electropores occurred. A delta Vm of greater than 540 mV caused stable electropore formation. In addition, the efficacy of two surface active polymers as membrane sealing agents was tested. Either 1 mmol/L Poloxamer-188 (8.1 kDa) or 1 mmol/L neutral dextran (10.1 kDa) prevented Mg2+ influx after delta Vms greater than 540 mV (p < 0.001, n = 7). These results suggest that the fields produced in victims of electric shock are sufficient to damage cell membranes by a nonthermal mechanism and that nonionic surfactants may rapidly seal electropores. PMID- 8245108 TI - Experience with patients with burns who refuse blood transfusion for religious reasons. AB - Patients with extensive full-thickness burns who refuse blood transfusion present a challenge to the burn team. We have recently treated four such patients, and we achieved a successful outcome in three. Staged excision and skin graft operations with minimal blood loss were performed. Two patients were treated with erythropoietin. Patient autonomy should be accepted by the burn team, but opportunity should exist for reassessment of treatment plans. PMID- 8245109 TI - Preliminary evaluation of vibriolysin, a novel proteolytic enzyme composition suitable for the debridement of burn wound eschar. AB - Vibriolysin, a proteolytic enzyme secreted by the marine microorganism Vibrio proteolyticus, was evaluated for its efficacy as an enzymatic debridement agent. Initial in vitro experiments revealed that the protease readily hydrolyzed proteinaceous components of eschar (e.g., fibrin, elastin, and in particular, collagen). Enzyme selectivity towards the digestion of denatured proteins but not viable elements was demonstrated in vitro and in vivo by treatment of full thickness burn wounds with a porcine model. The full-thickness wound eschar was rapidly hydrolyzed by a hydrophilic vibriolysin composition with a resultant wound bed that appeared pink and viable. Vibriolysin exhibited desirable properties heretofore not described for the enzymatic debridement agents, in particular, its selective hydrolysis of dead but not viable tissue, debridement in the absence of bleeding, compatibility with adjunct therapies, and its unique shelf-life stability in a hydrophilic composition at ambient temperature. PMID- 8245110 TI - Arterial occlusion and progressive gangrene caused by mucormycosis in a patient with burns. AB - Ischemic necrosis of the upper extremities caused by invasive mucormycosis developed in a patient with soil contamination of severe burn wounds. An arteriogram of the arm showed complete obstruction of blood flow in the forearm. Histologic specimens showed nonseptate branching hyphae obliterating the arterial lumens. Cutaneous mucormycosis affects patients who are immunocompromised, including victims of multiple trauma and burns. This case represents a previously undescribed clinical presentation in a patient with major burns. Because of its lethal nature, mucormycosis in a patient with burns must be treated with aggressive surgical debridement, including amputation, and with parentral amphotericin B at the earliest sign of cutaneous presence. PMID- 8245111 TI - An effective technique of scrotal harvest. AB - In male patients with extensive burns, the scrotum is often spared and is potentially a very useful donor site. We describe a technique that simplifies scrotal harvest and facilitates the procurement of clinically useful amounts of split-thickness skin. PMID- 8245112 TI - The elderly patient with burns: treatment considerations. AB - When therapists treat elderly patients with burns, they should ask themselves several questions: Am I treating elderly patients with burns "just like other patients with burns?" Am I assessing the patient's joint mobility and strength as critically as I should? Am I ensuring that further tissue trauma is avoided by instructing the nursing staff in turning/transfer techniques? Am I routinely monitoring heart rate and blood pressure? Am I requiring repetition and practice of exercises before the patient's discharge? Is my patient safe to function in the environment to which he or she is being discharged? Most physical changes in the elderly population are attributed to the aging process but often are due to diminished movement disorders arising from poor physical fitness and disuse. The "slowdown" in the older adult may result from a lack of appropriate movement practice and failure to receive reinforcement for activity. Regardless of age, physiological challenges result in enhanced functional status. Training for a specific skill increases performance in that skill just as strength training increases strength. Practice or repetition of a task improves movement and response time for that task. Appropriate physical training programs for the elderly can (1) counterbalance age-associated declines in work capacity and physical performance; (2) maintain and improve range of motion, strength, flexibility, balance, and endurance; and (3) reduce the disability of the cardiovascular, respiratory, and musculoskeletal systems that may result in decreased risk of falls and accidents. Practice of a task and maintenance of physical fitness helps ensure a high level of motor performance and speed as a person grows older. One is never too old to exercise! PMID- 8245113 TI - Conceptual and methodologic problems in the evaluation of a new burn treatment modality. AB - This article describes and analyzes major conceptual and methodologic problems that occurred during the implementation and evaluation of a new home-based burn treatment modality. Problems described include those of conceptualization, measurement, patient sampling, and follow-up, data collection, and analysis. The origins of each specific problem are discussed along with their effects on the implementation of the project and project outcomes. Solutions for each set of problems are suggested. PMID- 8245114 TI - Resting metabolic rate variability as influenced by mouthpiece and noseclip practice procedures. AB - Mouthpiece and noseclip introduction affect resting physiologic function (i.e., hyperventilation, anxiety), which can influence a resting metabolic rate measurement. It also is obvious that practice is required to minimize this influence when such apparatuses are used. However, the type of practice that best minimizes this influence has not been established. To determine the effect of acute and longer term practice on measurement variability of resting metabolic rate, 27 healthy, premenopausal women were randomly assigned to one of three groups: (1) a practice group, involving three 10-minute practice sessions 1 day to 1 week before resting metabolic rate was measured (n = 9); (2) an acclimation group, in which each subject acclimated to the mouthpiece and noseclip for 5 minutes before the resting metabolic rate measurement (n = 8); and (3) a control group (no practice) (n = 10). Resting metabolic rate was measured three times for each subject via a 5-minute oxygen uptake in the early morning (5:00 to 8:30 am) after a 25-minute supine rest. Resting metabolic rate measurements (kJ/day) and intraindividual variance estimates, adjusted for age and weight, were calculated to determine which practice group had the lowest resting metabolic rate measurement and the lowest variability around each individual's repeated resting metabolic rate measures. Of the three averaged resting metabolic rate measures, significantly lower 24-hour energy expenditures were found with the practice (5647 kJ) and acclimation (5550 kJ) groups when compared with those of the control group (6077 kJ) (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245115 TI - National Burn Awareness Week: how are burn care facilities in the United States participating? AB - A survey was conducted from burn care facilities (BCF) regarding their participation in National Burn Awareness Week (NBAW). The second week in February has been proclaimed NBAW. This is the only national campaign of this scope and is supported by the American Burn Association. The NBAW Task Force meets annually to compose and review the burn awareness materials that are sent to all BCF. A telephone survey was conducted, and responses to questions were obtained from all 148 of the BCF listed in the American Burn Association's "Burn Care Resources in North America 1991-1992." The purpose of the study was to answer questions regarding BCF involvement with NBAW and burn prevention programs. Review of the literature reveals this to be the first national study to elicit this type of information. Information sharing of this magnitude can have a dynamic effect on the participation of BCF in NBAW and can result in increased burn awareness in the BCF's local communities. An added benefit is that the task force receives specific input direct from those using the campaign, which validates the continuation of NBAW and the support of the ABA. PMID- 8245116 TI - Submembranous junctional plaque proteins include potential tumor suppressor molecules. PMID- 8245117 TI - Nuclear organization of splicing snRNPs during differentiation of murine erythroleukemia cells in vitro. AB - Murine erythroleukemia (MEL) cells are erythroid progenitors that can be induced to undergo terminal erythroid differentiation in culture. We have used MEL cells here as a model system to study the nuclear organization of splicing snRNPs during the physiological changes in gene expression which accompany differentiation. In uninduced MEL cells, snRNPs are widely distributed throughout the nucleoplasm and show an elevated concentration in coiled bodies. Within the first two days after induction of terminal erythroid differentiation, the pattern of gene expression changes, erythroid-specific transcription is activated and transcription of many other genes is repressed. During this early stage splicing snRNPs remain widely distributed through the nucleoplasm and continue to associate with coiled bodies. At later stages of differentiation (four to six days), when total transcription levels have greatly decreased, splicing snRNPs are redistributed. By six days postinduction snRNPs were concentrated in large clusters of interchromatin granules and no longer associated with coiled bodies. At the end-point of erythroid differentiation, just before enucleation, we observe a dramatic segregation of splicing snRNPs from the condensed chromatin. Analysis by EM shows that the snRNPs are packaged into a membrane-associated structure at the nuclear periphery which we term the "SCIM" domain (i.e., SnRNP Clusters Inside a Membrane). PMID- 8245118 TI - The structure of replicating kinetoplast DNA networks. AB - Kinetoplast DNA (kDNA), the mitochondrial DNA of Crithidia fasciculata and related trypanosomatids, is a network containing approximately 5,000 covalently closed minicircles which are topologically interlocked. kDNA synthesis involves release of covalently closed minicircles from the network, and, after replication of the free minicircles, reattachment of the nicked or gapped progeny minicircles to the network periphery. We have investigated this process by electron microscopy of networks at different stages of replication. The distribution of nicked and closed minicircles is easily detectable either by autoradiography of networks radiolabeled at endogenous nicks by nick translation or by twisting the covalently closed minicircles with intercalating dye. The location of newly synthesized minicircles within the network is determined by autoradiography of network is determined by autoradiography of networks labeled in vivo with a pulse of [3H]thymidine. These studies have clarified structural changes in the network during replication, the timing of repair of nicked minicircles after replication, and the mechanism of division of the network. PMID- 8245119 TI - Multiple regions of NSR1 are sufficient for accumulation of a fusion protein within the nucleolus. AB - NSR1, a 67-kD nucleolar protein, was originally identified in our laboratory as a nuclear localization signal binding protein, and has subsequently been found to be involved in ribosome biogenesis. NSR1 has three regions: an acidic/serine-rich NH2 terminus, two RNA recognition motifs, and a glycine/arginine-rich COOH terminus. In this study we show that NSR1 itself has a bipartite nuclear localization sequence. Deletion of either basic amino acid stretch results in the mislocation of NSR1 to the cytoplasm. We further demonstrate that either of two regions, the NH2 terminus or both RNA recognition motifs, are sufficient to localize a bacterial protein, beta-galactosidase, to the nucleolus. Intensive deletion analysis has further defined a specific acidic/serine-rich region within the NH2 terminus as necessary for nucleolar accumulation rather than nucleolar targeting. In addition, deletion of either RNA recognition motif or point mutations in one of the RNP consensus octamers results in the mislocalization of a fusion protein within the nucleus. Although the glycine/arginine-rich region in the COOH terminus is not sufficient to bring beta-galactosidase to the nucleolus, our studies show that this domain is necessary for nucleolar accumulation when an RNP consensus octamer in one of the RNA recognition motifs is mutated. Our findings are consistent with the notion that nucleolar localization is a result of the binding interactions of various domains of NSR1 within the nucleolus rather than the presence of a specific nucleolar targeting signal. PMID- 8245120 TI - Targeting and mistargeting of plasma membrane adaptors in vitro. AB - Targeting and recruitment of the plasma membrane (PM) clathrin-coated vesicle adaptor complexes has been studied using an in vitro system based on permeabilized acceptor cells and donor cytosol. Through the use of species- and/or tissue-specific antibodies, only newly recruited exogenous PM adaptors are visualized. Targeting of PM adaptors can be switched from the plasma membrane to a perinuclear compartment by GTP gamma S or excess calcium. Prior treatment with brefeldin A prevents GTP gamma S-induced mistargeting. Double-labeling immunofluorescence and immunogold EM indicate that the perinuclear PM adaptor binding compartment is late endosomal. We propose that receptors for PM adaptors cycle between the plasma membrane and an endosomal storage compartment. Normally the receptors would be switched on only at the plasma membrane, but both GTP gamma S and calcium are capable of reversing this switch. Intracellular sequestration of PM adaptor receptors may provide the cell with a mechanism for up-regulating endocytosis following a burst of exocytosis. PMID- 8245121 TI - Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. AB - The clathrin-coated pit lattice is held onto the plasma membrane by an integral membrane protein that binds the clathrin AP-2 subunit with high affinity. In vitro studies have suggested that this protein controls the assembly of the pit because membrane bound AP-2 is required for lattice assembly. If so, the AP-2 binding site must be a resident protein of the coated pit and recycle with other receptors that enter cells through this pathway. Proper recycling, however, would require the switching off of AP-2 binding to allow the binding site to travel through the endocytic pathway unencumbered. Evidence for this hypothesis has been revealed by the cationic amphiphilic class of drugs (CAD), which have previously been found to inhibit receptor recycling. Incubation of human fibroblasts in the presence of these drugs caused clathrin lattices to assemble on endosomal membranes and at the same time prevented coated pit assembly at the cell surface. These effects suggest that CADs reverse an on/off switch that controls AP-2 binding to membranes. We conclude that cells have a mechanism for switching on and off AP-2 binding during the endocytic cycle. PMID- 8245122 TI - The subcellular distribution of early endosomes is affected by the annexin II2p11(2) complex. AB - The tyrosine kinase substrate annexin II is a member of a multigene family of Ca2+ and lipid-binding proteins which have been implicated in a number of membrane-related events. We have analyzed the subcellular distribution of annexin II in relation to other cellular components in normal and specifically manipulated MDCK cells. In a polarized monolayer of MDCK cells annexin II and its cellular ligand p11 are restricted almost exclusively to the cortical regions of the cells which also contain peripheral early endosomes. Treatment of the polarized cells with low Ca2+ medium leads to a disintegration of the cortical cytoskeleton and a translocation of both, the annexin II2p11(2) complex and early endosomes, to the cytoplasm. A similar translocation which is however specific for the annexin II2p11(2) complex and early endosomes and does not affect other elements of the cell cortex is observed in cells expressing a trans-dominant annexin II-p11 mutant. This chimeric mutant protein causes the aggregation of endogenous annexin II and p11 and the simultaneous detachment of early endosomes from the cell periphery resulting in the binding of the early endosomes but no other components of the endocytotic or biosynthetic pathways to the annexin II/p11 aggregates. The specificity of this effect argues for the association of the annexin II2p11(2) complex with early endosomes and suggests that this association contributes to establish the peripheral localization of early endosomal structures. PMID- 8245123 TI - Mutational and secondary structural analysis of the basolateral sorting signal of the polymeric immunoglobulin receptor. AB - The 17-juxtamembrane cytoplasmic residues of the polymeric immunoglobulin receptor contain an autonomous basolateral targeting signal that does not mediate rapid endocytosis (Casanova, J. E., G. Apodaca, and K. E. Mostov. Cell. 66:65 75). Alanine-scanning mutagenesis identifies three residues in this region, His656, Arg657, and Val660, that are most essential for basolateral sorting and two residues, Arg655 and Tyr668, that play a lesser role in this process. Progressive truncations suggested that Ser664 and Ile665 might also play a role in basolateral sorting. However, mutation of these residues to Ala or internal deletions of these residues did not affect basolateral sorting, indicating that these residues are probably not required for basolateral sorting. Two-dimensional NMR spectroscopy of a peptide corresponding to the 17-mer signal indicates that the sequence Arg658-Asn-Val-Asp661 has a propensity to adopt a beta-turn in solution. Residues COOH-terminal to the beta-turn (Arg662 to Arg669) seem to take up a nascent helix structure in solution. Substitution of Val660 with Ala destabilizes the turn, while mutation of Arg657 to Ala does not appear to affect the turn structure. Neither mutation detectably altered the stability of the nascent helix in the COOH-terminal portion of the peptide. PMID- 8245124 TI - Triad formation: organization and function of the sarcoplasmic reticulum calcium release channel and triadin in normal and dysgenic muscle in vitro. AB - Excitation-contraction (E-C) coupling is thought to involve close interactions between the calcium release channel (ryanodine receptor; RyR) of the sarcoplasmic reticulum (SR) and the dihydropyridine receptor (DHPR) alpha 1 subunit in the T tubule membrane. Triadin, a 95-kD protein isolated from heavy SR, binds both the RyR and DHPR and may thus participate in E-C coupling or in interactions responsible for the formation of SR/T-tubule junctions. Immunofluorescence labeling of normal mouse myotubes shows that the RyR and triadin co-aggregate with the DHPR in punctate clusters upon formation of functional junctions. Dysgenic myotubes with a deficiency in the alpha 1 subunit of the DHPR show reduced expression and clustering of RyR and triadin; however, both proteins are still capable of forming clusters and attaining mature cross-striated distributions. Thus, the molecular organization of the RyR and triadin in the terminal cisternae of SR as well as its association with the T-tubules are independent of interactions with the DHPR alpha 1 subunit. Analysis of calcium transients in dysgenic myotubes with fluorescent calcium indicators reveals spontaneous and caffeine-induced calcium release from intracellular stores similar to those of normal muscle; however, depolarization-induced calcium release is absent. Thus, characteristic calcium release properties of the RyR do not require interactions with the DHPR; neither do they require the normal organization of the RyR in the terminal SR cisternae. In hybrids of dysgenic myotubes fused with normal cells, both action potential-induced calcium transients and the normal clustered organization of the RyR are restored in regions expressing the DHPR alpha 1 subunit. PMID- 8245125 TI - Yeast actin with a mutation in the "hydrophobic plug" between subdomains 3 and 4 (L266D) displays a cold-sensitive polymerization defect. AB - Holmes et al. (Holmes, K. C., D. Popp, W. Gebhard, and W. Kabsch. 1990. Nature [Lond.] 347: 44-49) hypothesized that between subdomains 3 and 4 of actin is a loop of 10 amino acids including a four residue hydrophobic plug that inserts into a hydrophobic pocket formed by two adjacent monomers on the opposing strand thereby stabilizing the F-actin helix. To test this hypothesis we created a mutant yeast actin (L266D) by substituting Asp for Leu266 in the plug to disrupt this postulated hydrophobic interaction. Haploid cells expressing only this mutant actin were viable with no obvious altered phenotype at temperatures above 20 degrees C but were moderately cold-sensitive for growth compared with wild type cells. The critical concentration for polymerization increased 10-fold at 4 degrees C compared with wild-type actin. The length of the nucleation phase of polymerization increased as the temperature decreased. At 4 degrees C nucleation was barely detectable. Addition of phalloidin-stabilized F-actin nuclei and phalloidin restored L266D actin's ability to polymerize at 4 degrees C. This mutation also affects the overall rate of elongation during polymerization. Small effects of the mutation were observed on the exchange rate of ATP from G-actin, the G-actin intrinsic ATPase activity, and the activation of myosin S1 ATPase activity. Circular dichroism measurements showed a 15 degrees C decrease in melting temperature for the mutant actin from 57 degrees C to 42 degrees C. Our results are consistent with the model of Holmes et al. (Holmes, K. C., D. Popp, W. Gebhard, and W. Kabsch. 1990. Nature [Lond.]. 347:44-49) involving the role of the hydrophobic plug in actin filament stabilization. PMID- 8245126 TI - Peripherin expression in hippocampal neurons induced by muscle soluble factor(s). AB - Previous studies have shown that neuronal cells in culture can switch neurotransmitters when grown in the presence of different target cells. To examine whether this plasticity extends to structural proteins, we cocultured hippocampal neurons and pituitary-derived neuroendocrine (AtT20) cells with astrocytes, kidney epithelial cells, or skeletal muscle cells. As a marker of phenotypic change we used the cytoskeletal protein peripherin, a type III intermediate filament (IF) subunit which is not expressed in hippocampal neurons and AtT20 cells. We show here that soluble factor(s) secreted specifically from skeletal muscle cells can induce the expression and de novo assembly of peripherin in a subset of post-mitotic neurons. We further demonstrate that one of these factors is the Leukemia Inhibitory Factor/Cholinergic Neuronal Differentiation Factor. The environmentally regulated expression of peripherin implies a remarkable degree of plasticity in the cytoskeletal organization of postmitotic CNS cells and provides a noninvasive model system to examine the de novo assembly of IF proteins under in vivo conditions. PMID- 8245127 TI - Axonal regulation of Schwann cell integrin expression suggests a role for alpha 6 beta 4 in myelination. AB - Ensheathment and myelination of axons by Schwann cells in the peripheral nervous system requires contact with a basal lamina. The molecular mechanism(s) by which the basal lamina promotes myelination is not known but is likely to reflect the activity of integrins expressed by Schwann cells. To initiate studies on the role of integrins during myelination, we characterized the expression of two integrin subunits, beta 1 and beta 4, in an in vitro myelination system and compared their expression to that of the glial adhesion molecule, the myelin-associated glycoprotein (MAG). In the absence of neurons, Schwann cells express significant levels of beta 1 but virtually no beta 4 or MAG. When Schwann cells are cocultured with dorsal root ganglia neurons under conditions promoting myelination, expression of beta 4 and MAG increased dramatically in myelinating cells, whereas beta 1 levels remained essentially unchanged. (In general agreement with these findings, during peripheral nerve development in vivo, beta 4 levels also increase during the period of myelination in sharp contrast to beta 1 levels which show a striking decrease.) In cocultures of neurons and Schwann cells, beta 4 and MAG appear to colocalize in nascent myelin sheaths but have distinct distributions in mature sheaths, with beta 4 concentrated in the outer plasma membrane of the Schwann cell and MAG localized to the inner (periaxonal) membrane. Surprisingly, beta 4 is also present at high levels with MAG in Schmidt Lanterman incisures. Immunoprecipitation studies demonstrated that primary Schwann cells express beta 1 in association with the alpha 1 and alpha 6 subunits, while myelinating Schwann cells express alpha 6 beta 4 and possibly alpha 1 beta 1. beta 4 is also downregulated during Wallerian degeneration in vitro, indicating that its expression requires continuous Schwann cell contact with the axon. These results indicate that axonal contact induces the expression of beta 4 during Schwann cell myelination and suggest that alpha 6 beta 4 is an important mediator of the interactions of myelinating Schwann cells with the basal lamina. PMID- 8245128 TI - ATP-dependent degradation of a mutant serine: pyruvate/alanine:glyoxylate aminotransferase in a primary hyperoxaluria type 1 case. AB - Primary hyperoxaluria type 1 (PH 1), an inborn error of glyoxylate metabolism characterized by excessive synthesis of oxalate and glycolate, is caused by a defect in serine:pyruvate/alanine:glyoxylate aminotransferase (SPT/AGT). This enzyme is peroxisomal in human liver. Recently, we cloned SPT/AGT-cDNA from a PH 1 case, and demonstrated a point mutation of T to C in the coding region of the SPT/AGT gene encoding a Ser to Pro substitution at residue 205 (Nishiyama, K., T. Funai, R. Katafuchi, F. Hattori, K. Onoyama, and A. Ichiyama. 1991. Biochem. Biophys. Res. Commun. 176:1093-1099). In the liver of this patient, SPT/AGT was very low with respect to not only activity but also protein detectable on Western blot and immunoprecipitation analyses. Immunocytochemically detectable SPT/AGT labeling was also low, although it was detected predominantly in peroxisomes. On the other hand, the level of translatable SPT/AGT-mRNA was higher than normal, indicating that SPT/AGT had been synthesized in the patient's liver at least as effectively as in normal liver. Rapid degradation of the mutant SPT/AGT was then demonstrated in transfected COS cells and transformed Escherichia coli, accounting for the low level of immunodetectable mutant SPT/AGT in the patient's liver. The mutant SPT/AGT was also degraded much faster than normal in an in vitro system with a rabbit reticulocyte extract, and the degradation in vitro was ATP dependent. These results indicate that a single amino acid substitution in SPT/AGT found in the PH1 case leads to a reduced half-life of this protein. It appears that the mutant SPT/AGT is recognized in cells as an abnormal protein to be eliminated by degradation. PMID- 8245129 TI - The mechanism for the activation of latent TGF-beta during co-culture of endothelial cells and smooth muscle cells: cell-type specific targeting of latent TGF-beta to smooth muscle cells. AB - Transforming growth factor-beta (TGF-beta) is secreted in a latent form and activated during co-culture of endothelial cells and smooth muscle cells. Plasmin located on the surface of endothelial cells is required for the activation of latent TGF-beta (LTGF-beta) during co-culture, and the targeting of LTGF-beta to the cellular surface is requisite for its activation. In the present study, the cellular targeting of LTGF-beta was examined. We detected the specific binding of 125I-large LTGF-beta 1 isolated from human platelets to smooth muscle cells but not to endothelial cells. A mAb against the latency-associated peptide (LAP) of large LTGF-beta 1 complex, which blocked the binding of 125I-large LTGF-beta 1 to smooth muscle cells, inhibited the activation of LTGF-beta during co-culture. The binding of 125I-large LTGF-beta 1 could not be competed either by mannose-6 phosphate (300 microM) or by the synthetic peptide Arg-Gly-Asp-Ser (300 micrograms/ml). These results indicate that the targeting of LTGF-beta to smooth muscle cells is required for the activation of LTGF-beta during co-culture of endothelial cells and smooth muscle cells. The targeting of LTGF-beta to smooth muscle cells is mediated by LAP, and the domain of LAP responsible for the targeting to smooth muscle cells may not be related to mannose-6-phosphate or an Arg-Gly-Asp sequence, both of which have been previously proposed as candidates for the cellular binding domains within LAP. PMID- 8245130 TI - Structure and expression of fibulin-2, a novel extracellular matrix protein with multiple EGF-like repeats and consensus motifs for calcium binding. AB - A new protein, fibulin-2, was predicted from sequence analysis of cDNA clones obtained from a mouse fibroblast library. This protein consists of a 1195-residue polypeptide preceded by a 26-residue signal peptide. The COOH-terminal region of 787 amino acids contained three anaphylatoxin-related segments (domain I), 11 EGF like repeats (domain II), 10 of which had a consensus motif for calcium-binding, and a 115-residue globular domain III. Except for two additional EGF-like repeats, this COOH-terminal region showed 43% sequence identity with the previously described fibulin-1 (BM-90). The NH2-terminal 408 residues, unique to fibulin-2, showed no sequence homology to other known proteins and presumably form two additional domains that differ in their cysteine content. Recombinant fibulin-2 was produced and secreted by human cell clones as a disulfide-bonded trimer. Rotary shadowing visualized the protein as three 40-45 nm long rods which are connected at one end in a globe-like structure. No significant immunological cross-reaction could be detected between fibulin-1 and fibulin-2. Production of the fibulin-2 was demonstrated by Northern blots and radioimmunoassay in fibroblasts but not in several tumor cell lines. Together with the observation that the serum level of fibulin-2 is 1,000-fold lower than that of fibulin-1, the data indicate that these two isoforms are not always coordinately expressed. This is also suggested by Northern blots of tissue mRNAs and by immunofluorescence localizations using mouse tissues. The latter studies also demonstrated an extracellular localization for fibulin-2 in basement membranes and other connective tissue compartments. PMID- 8245131 TI - Thrombin adhesive properties: induction by plasmin and heparan sulfate. AB - We have previously demonstrated that chemically modified thrombin preparations induce endothelial cell (EC) adhesion, spreading and cytoskeletal reorganization via an Arg-Gly-Asp (RGD) sequence and the alpha v beta 3 integrin. Native thrombin, however, did not exhibit adhesive properties, consistent with crystal structure analysis, showing that Gly-Asp residues of the RGD epitope are buried within the molecule. We have now identified a possible physiological mean of converting thrombin to an adhesive protein. Plasmin, the major end product of the fibrinolytic system, converted thrombin to an adhesive protein for EC in a time and dose-dependent manner. EC adhesion and spreading was also induced by a low molecular weight (approximately 3,000 D) cleavage fragment generated upon incubation of thrombin with plasmin. Cell adhesion mediated by this fragment was completely inhibited by the synthetic peptide GRGDSP. Conversion of thrombin to an adhesive molecule was significantly enhanced in the presence of heparin or heparan sulfate, while other glycosaminoglycans (GAGs) (e.g., dermatan sulfate, keratan sulfate, chondroitin sulfate) had no effect. The role of cell surface heparan sulfate in thrombin conversion to EC adhesive protein was investigated using CHO cell mutants defective in various aspects of GAG synthesis. Incubation of both thrombin and a suboptimal amount of plasmin on the surface of formaldehyde fixed wild-type CHO-KI cells resulted in an efficient conversion of thrombin to an adhesive molecule, as indicated by subsequent induction of EC attachment. In contrast, there was no effect to incubation of thrombin and plasmin with fixed CHO mutant cells lacking both heparan sulfate and chondroitin sulfate, or with cells expressing no heparan sulfate and a three-fold increase in chondroitin sulfate. A similar gain of adhesive properties was obtained upon incubation of thrombin and plasmin in contact with native, but not heparinase treated extracellular matrix (ECM) produced by cultured ECs. It appears that cell surface and ECM-associated heparan sulfate modulate thrombin adhesive properties through its heparin binding site in a manner that enables suboptimal amounts of plasmin to expose the RGD domain. Our results demonstrate, for the first time, a significant modulation of thrombin molecule by heparin, resulting in its conversion to a potent adhesive protein for ECs. This conversion is most effective in contact with cell surfaces, basement membranes and ECM. PMID- 8245133 TI - Abnormal shyness in children. AB - Recent nonclinical studies of children who do not often interact with their peers have identified at least three different kinds of solitude: temperamental shyness, social-evaluative shyness, and unsociability. These kinds of solitude can be clearly distinguished from social nonacceptance. The evidence for the different faces of children's solitude and their situational, age, and cultural specificity is reviewed, and potential implications for clinical child psychology are discussed. Clinical practitioners can profit from a recognition of the multifaceted nature of solitude in childhood. PMID- 8245132 TI - Sequence and tissue distribution of the integrin alpha 9 subunit, a novel partner of beta 1 that is widely distributed in epithelia and muscle. AB - The integrin family of adhesion receptors consists of several heterodimeric glycoproteins, each composed of one alpha and one beta subunit. A novel integrin alpha subunit partial cDNA isolated from TGF-beta stimulated guinea pig airway epithelial cells has previously been reported (Erle, D.J., D. Sheppard, J. Bruess, C. Ruegg, and R. Pytela. 1991. Am. J. Respir. Cell Mol. Biol. 5:170-177). We have now determined cDNA and amino acid sequence for the human homolog of this subunit, named alpha 9, from a human lung cDNA library, a human small intestine cDNA library, and cDNA from the cell lines U937, HL-60 and Tera-2. This sequence is predicted to encode a 1006-amino acid mature protein that shares 39% identity with the previously identified integrin subunit alpha 4. By Northern blot analysis, alpha 9 mRNA was detected in the human carcinoma cell lines Tera-2 and Caco-2. Anti-peptide antibodies against the predicted COOH-terminal sequence of alpha 9 immunoprecipitated a heterodimer (140 kD/115 kD nonreduced; 150 kD/130 kD reduced) from Tera-2 lysates. Immunodepletion of beta 1-containing integrins with Tera-2 lysates removed alpha 9 immunoreactivity, suggesting that beta 1 is the principal beta subunit partner for alpha 9 in these cells. alpha 9 was detected by immunohistochemistry in airway epithelium, in the basal layer of squamous epithelium, and in smooth muscle, skeletal muscle, and hepatocytes. PMID- 8245134 TI - Depressed mothers' speech to their infants and its relation to infant gender and cognitive development. AB - The speech of depressed and well mothers during play with their infants at two months was compared on dimensions of structure and semantics. No differences between maternal groups were found on measures of complexity and syntax. However, the speech of depressed women expressed more negative affect, was less focused on infant experience, and tended to show less acknowledgement of infant agency. Speech style of depressed women also varied according to infant gender. Regression analyses indicated that the quality of maternal communication with the infant, and particularly the focus of speech, mediated the association between depression and infant cognitive development in the first 18 months. PMID- 8245135 TI - Depression in 11-16-year-old girls: the role of past parental psychopathology and exposure to recent life events. AB - Interviews with parents of a non-referred sample of 11-16-year-old girls (n = 82) revealed that a significantly greater proportion of mothers with a lifetime history of any psychiatric disorder also reported one or more recent undesirable life events focused on the adolescent compared with mothers with no such history. Lifetime episodes of maternal depression and recent undesirable life events exerted significant additive effects on the likelihood of depression occurring in the previous 12 months in adolescent girls. Some families may be "life event prone" as a consequence of lifetime episodes of parental psychopathology. PMID- 8245136 TI - Depressive symptomatology, perceptions of attractiveness, and body image in children. AB - Fifty-seven children, aged 9-12 completed the Children's Depression Inventory, rated their bodies on dimensions of attractiveness, effectiveness and vulnerability, and rated themselves and three other children on global attractiveness. Dysphoria was significantly correlated with both measures of dissatisfaction of own attractiveness. No significant correlations occurred between CDI scores and effectiveness or vulnerability. Stepwise regression showed dysphoria to be predicted by dissatisfaction with the body's attractiveness and by age with the former being the most powerful. No subject differences were found on rating the attractiveness of other children. Results are compared to research with adult subjects. PMID- 8245137 TI - Self-esteem, parental appraisal and body size in children. AB - This study investigates the influence of body size, parental appraisal of body size, and children's beliefs about parental appraisal, on self-esteem in children from 9 to 11 years old. Parents' and children's responses to a matched question about body size suggest that children are accurate predictors of parental evaluation and that their self-esteem scores are influenced both by actual parental dissatisfaction and beliefs about parental dissatisfaction. For boys, lower self-esteem is associated both with thinness and being perceived as too thin. For girls, lower self-esteem is more associated with fatness. PMID- 8245138 TI - Hyperactivity and spelling disability: testing for shared genetic aetiology. AB - The influence of genetic factors in the comorbidity of spelling disability and hyperactivity was investigated in two samples of 190 and 260 same sex twin pairs. The method of bivariate group heritability was used to estimate the genetic correlation for spelling disability and hyperactivity. A similar though not statistically significant value for the genetic correlation was obtained for the two samples (0.29 and 0.42). It was estimated that approximately 75% of the co occurrence of these two conditions was due to shared genetic influences. PMID- 8245139 TI - Adolescent adjustment to chronic physical disorders--I. Comparing neurological and non-neurological conditions. AB - Early research on child adjustment to chronic illness assumed that each condition had a unique impact. Recently researchers have suggested that all chronic conditions influence adjustment in similar ways. To compare these models, data were collected on 165 adolescents having chronic conditions with and without brain involvement, and 49 healthy controls. Adolescents with brain-based conditions had more behaviour problems, less autonomous functioning and poorer school achievement. Children with conditions having no brain involvement differed from controls only in reporting less work experience and having lower math achievement scores. These findings support a modified perspective that involves both general factors and effects specific to brain-based conditions. PMID- 8245140 TI - Psychotic disorders diagnosed in adolescence. Outcome at age 30 years. AB - Fifty-five cases who had been diagnosed in adolescence as suffering from a psychotic condition while attending a child or adult psychiatric clinic were compared with age-, sex- and school-matched cases with respect to various outcome measures at age 30 years. Overall, the outcome for young people who need psychiatric care for psychotic conditions in adolescence appears to be gloomy with respect to psychosocial adjustment. There were clear differences in outcome depending on specific psychosis diagnosis category at the time of original diagnosis. Females tended to have better outcome than males, but differences were small and not conclusive in this respect. PMID- 8245141 TI - DSM-III-R disorders, social factors and management of school attendance problems in the normal population. AB - Eighty 13-15-year-old children who failed to attend one of four schools for more than 40% of a term, without good reason, were studied. A systematic schedule (C.A.P.A.) was used in interviewing parents and children. Twenty-five had DSM-III R Disruptive Behaviour Disorders and 15 had Anxiety/Mood Disorders. Truancy was associated with the former and school refusal with the latter but both often occurred without any Disorder. Fourteen children had neither school refusal nor truancy. Compared to controls, poor attenders came from materially disadvantaged homes. School refusal with anxiety disorders rarely received psychiatric treatment. Non-disturbed absentees were not usually dealt with appropriately. PMID- 8245142 TI - Chronic family adversity and infant attachment security. AB - A beginning step in the prevention of psychopathology in children is the identification of conditions and events associated with a disproportionately high incidence of behavior problems. Rutter, Cox, Tupling, Berger and Yule (1975a, British Journal of Psychiatry, 126, 493-509) have reported a dramatic increase in the probability of children's adjustment difficulties as a function of multiple family stressors. However, this association has never been tested with infants. Among instruments of infant behavior, attachment classification has been found to be a significant predictor of later adjustment problems, particularly among low income samples. The present investigation examines the relation between six significant familial stressors and infant attachment security in a sample of 100 low-income parent-infant dyads. Family stressors included parental criminality, maternal depressive symptomatology, maternal personality risk, overcrowding in the home, and the quality of the relationship with a significant other. Cumulative family adversity was found to differentiate secure from insecure infants, but only among families with three or four stressors present. There also was moderate support for one hypothesis derived from attachment theory, namely that stressors more closely associated with maternal functioning are more common among families with insecurely attached infants. Conceptual links between stressors and attachment are discussed, including pathways by which chronic stressors may interfere with the formation and maintenance of secure mother infant attachment relations. PMID- 8245143 TI - Is the Continuous Performance Task a valuable research tool for use with children with Attention-Deficit-Hyperactivity Disorder? AB - The Continuous Performance Task (CPT) has become a popular research tool used to distinguish children with Attention-Deficit-Hyperactivity Disorder (ADHD) from their normal peers. Whether hyperactive children have a vigilance decrement remains an ongoing controversy. The theoretical basis of the CPT and variables known to influence performance are examined in order to interpret the inconsistencies evident in the research. Studies which employed the CPT in order to examine the possibility of sustained attention deficits in children with ADHD will be reviewed. The results are examined in light of group selection criteria, task variables and situational and external variables. It is concluded that there is no compelling evidence for a sustained deficit in ADHD children. An alternative theoretical model for understanding the results of CPT performance in ADHD children is provided. PMID- 8245144 TI - Convergence of the Child Behavior Checklist with structured interview-based psychiatric diagnoses of ADHD children with and without comorbidity. AB - We evaluated the convergence of CBCL scales with the diagnosis of ADHD and comorbid disorders in 133 ADHD and 118 normal control boys, aged 6-17 years old. We evaluated the strength of association between each CBCL scale and structured interview derived diagnoses with Total Predictive Value (TPV) and the odds-ratio (OR). Excellent convergence was found between the CBCL Attention Problems scale with the diagnosis of ADHD, between the Delinquent Behavior scale and the diagnosis of CD, and between the Anxiety/Depression scale and the diagnoses of Anxiety Disorders. These findings indicate that the CBCL could serve as a rapid and useful screening instrument to identify comorbid and non-comorbid cases of ADHD. PMID- 8245145 TI - The validity of the Five-Minute Speech Sample as an index of Expressed Emotion in parents of eating disorder patients. AB - This article examines the concurrent validity of the Five-Minute Speech Sample (FMSS) as an index of Expressed Emotion in a Dutch sample of 84 parents of adolescents suffering from anorexia or bulimia nervosa. The Camberwell Family Interview (CFI), the criterion measure of EE, and the FMSS were conducted on the same day. The levels of Expressed Emotion in these families were low when compared with the EE ratings from the schizophrenia studies. The FMSS and CFI-EE ratings showed a limited degree of overlap. Whether the limited association between the two methods is due to the low levels of criticism in our sample, to cultural differences and/or to differences in the psychopathology under study remains unclear. PMID- 8245146 TI - 2-(4-Hydrazinocarbonylphenyl)-4,5-diphenylimidazole as a versatile fluorescent derivatization reagent for the high-performance liquid chromatographic analysis of free fatty acids. AB - A new fluorescent derivatization reagent with a lophine skeleton, 2-(4 hydrazinocarbonylphenyl)-4,5-diphenylimidazole (HCPI), has been synthesized and applied to the assay of saturated free fatty acids. The HCPI derivatives with five representative saturated fatty acids [caproic acid (C6), lauric acid (C12), palmitic acid (C16), stearic acid (C18) and arachidic acid (C20)] were synthesized to examine their fluorescence properties. The fluorescence spectra (lambda ex ca. 335 nm, lambda em ca. 455 nm) of the derivatives were almost identical in methanol, n-heptane and acetonitrile. For the high-performance liquid chromatographic assay, fatty acids were derivatized with HCPI using a condensing agent, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, and pyridine at room temperature. The HCPI derivatives were separated on a C18 column with a gradient elution of methanol-water, and the eluates were monitored at 455 nm using an excitation wavelength of 335 nm. By this method, seven kinds of free fatty acid (C6, C10, C12, C14, C16, C18 and C20) could be determined using margaric acid (C17) as an internal standard. Calibration curves were linear over the range 0.2-400 pmol per 20-microliters injection (r = 0.994-1.000). Relative standard deviations of the peak-height ratio for five replicate measurements of fatty acids (100 pmol per injection) were 0.7-2.6%. Detection limits were 7-57 fmol at a signal-to-noise ratio of 3. The method could be successfully applied to the determination of four saturated fatty acids (C12, C14, C16 and C18) in normal human serum. PMID- 8245147 TI - Electrochemical detection of cobalt in hair following on-column derivatization with 1-(2-pyridylazo)-2-naphthol-6-sulphonic acid and reversed-phase liquid chromatography. AB - A method for the specific determination of cobalt based on reversed-phase liquid chromatography with amperometric detection via on-column complex formation has been developed. A water-soluble chelating agent, 1-(2-pyridylazo)-2-naphthol-6 sulphonic acid (PAN-6S), is added to the mobile phase and aqueous cobalt solutions are injected directly into the column to form in situ the cobalt-PAN-6S chelate, which is then separated from other metal PAN-6S chelates and subjected to reductive amperometric detection at a moderate potential of -0.3 V. Because the procedure eliminates the interference of oxygen and depresses the electrochemical reduction of the mobile phase-containing ligand PAN-6S, by virtue of the quasi-reversible electrode process of the cobalt-PAN-6S complex, a low detection limit of 0.06 ng can be readily obtained. Interference effects were examined for sixteen common metal species, and at a 5- to 8000-fold excess by mass no obvious interference was observed. The feasibility of the method as an approach to the specific analysis of cobalt in a hair sample has been demonstrated. PMID- 8245148 TI - Determination of dopamine and its metabolites in microdialysates by capillary liquid chromatography with electrochemical detection. AB - A liquid chromatographic method with packed fused-silica capillary columns and amperometric detection for the determination of trace amounts of catecholamines in microdialysis samples has been developed. The analysis time for dopamine and the products of its metabolism does not exceed 10 min. The minimum detectable dopamine concentration in a 2-microliters sample is 0.05 microgram/l. The relative standard deviation in concentration determinations is +/- 0.03 (n = 10). The small sample volume required and the speed of analysis make it possible to analyse biologically important compounds with a rapidly changing concentration. PMID- 8245149 TI - Basal and stimulated cyclic adenosine 3',5'-monophosphate production in rat adenopituitary homogenates: direct determination by high-performance liquid chromatography. AB - A sensitive and reliable procedure to quantify cAMP in biological samples, without involving radioisotopes, is described. After incubation, the assay mixture was boiled for 4 min, centrifuged, filtered and directly injected onto a mu Bondapak C18 column (10 microns particle size; 150 mm x 3.9 mm I.D.). cAMP was resolved within 40 min in isocratic mode using a mobile phase composed of 96% of 0.01 M ammonium acetate, pH 6.6 and 4% of acetonitrile-water &70:30, v/v) at a flow-rate of 0.5 ml/min. Detection was monitored at 254 nm and quantification was achieved by peak surface integration. The lowest detection limit was 1.6 pmol (signal-to-noise ratio = 2.5). The potential of this assay to perform structure activity of bioactive peptides is illustrated with growth hormone-releasing factor in rat adenopituitary homogenates. PMID- 8245150 TI - Lyophilization can generate artifacts in chromatographic profiles of inositol phosphates. AB - Lyophilized extracts of [3H]inositol-labelled rat heart or renal tubule preparations were found to contain unidentified 3H-labelled compounds in addition to the inositol phosphates. The appearance of these labelled substances was caused by the presence in the extracts of compounds which bound [3H]inositol when lyophilized together with it. These studies demonstrate a previously undescribed source of [3H]inositol-labelled compounds which can complicate chromatographic profiles of inositol phosphates. These problems can be overcome either by not lyophilizing the samples or by lyophilizing in the presence of 0.3 M urea, which prevents the association with [3H]inositol and does not interfere with the chromatography. PMID- 8245151 TI - Improved method to extract and concentrate porphyrins from liver tissue for analysis by high-performance liquid chromatography. AB - A new method to extract and concentrate porphyrins from liver tissue for analysis by high-performance liquid chromatography is described. Porphyrins were extracted with acetonitrile-hydrochloric acid, concentrated on disposable octadecylsilyl columns, brought to dryness, and reconstituted in 1.2 M hydrochloric acid. Dried porphyrins were stable at -80 degrees C for at least three weeks, and porphyrins reconstituted in hydrochloric acid were stable for at least 20 h. Yields of uro-, heptacarboxyl-, hexacarboxyl-, pentacarboxyl- and coproporphyrin were > or = 90%. The method allows determination of porphyrin concentrations in small (10-100 mg) samples of liver tissue. PMID- 8245152 TI - Rapid and simple method for the determination of nitrite in synaptosomal superfusates by liquid chromatography with electrochemical detection. AB - A rapid and simple technique using reversed-phase high-performance liquid chromatography with electrochemical detection (HPLC-ED) was developed for the determination of nitrite. Standard solutions of sodium nitrite produced a linear current response over the concentration range 1.5-30 microM. The chromatographic peaks of sodium nitrite were superimposable with peaks obtained after injection into the chromatograph of supernatants from rat cerebellar synaptosomes. The concentrations of nitrite in synaptosomes measured with HPLC-ED were similar to those found with a widely used colorimetric procedure. Therefore, this new method may represent a useful means of measuring nitrite in biological samples. PMID- 8245153 TI - Reusability of Bond Elut Certify columns for the extraction of drugs from plasma. AB - The reusability of Bond Elut Certify columns for the extraction of toxicologically relevant drugs from plasma has been evaluated. Pentobarbital, hexobarbital, mepivacaine, trimipramine and clonazepam were selected as test drugs to represent various classes of drugs. The columns were regenerated immediately after an extraction by washing with methanol, hydrochloric acid (1%), water and methanol, sequentially. These regenerated columns were found to be reusable for plasma. More than 85% of the test drugs were recovered when the columns were used three times. However, the extraction power of the regenerated columns decreased slightly with the number of reuses, so that a column should not be used more than two or three times. PMID- 8245154 TI - Rapid high-performance liquid chromatographic assay of ethynyloestradiol in rabbit plasma. AB - A method for the determination of ethynyloestradiol in samples of rabbit plasma containing pentobarbital and heparin, the former used as an anaesthetic and the latter as an anticoagulant, has been developed. Quantification was carried out using a reversed-phase high-performance liquid chromatographic (HPLC) method in isocratic mode at room temperature, with electrochemical detection at an applied potential of +1 V vs. Ag/AgCl. Under these conditions, the retention time for ethynyloestradiol was ca. 2.9 min, the average recovery from plasma was 74.5%, and the limit of detection was 10 pg, corresponding to a plasma concentration of 50 pg/ml using 1 ml of plasma. Natural oestrogens, oestriol, oestradiol and oestrone showed peaks that did not interfere with ethynyloestradiol, and retention times of ca. 0.8, 2.4 and 3.4 min, respectively. PMID- 8245155 TI - Determination of mitomycin C in human aqueous humor and serum by high-performance liquid chromatography. AB - Mitomycin C (MMC) is used in the treatment of disseminated adenocarcinoma of the stomach and pancreas and is used in ophthalmology as adjunctive therapy in trabeculectomy. Since only small volumes of aqueous humor are available for analysis, a sensitive method requiring limited sample preparation was developed. An internal standard, 4-aminoacetophenone, was added to aqueous humor specimens, and the solution was directly injected into the high-performance liquid chromatographic (HPLC) column. The use of a short 50-mm C18 reversed-phase column gave adequate resolution of peaks with improved sensitivity. The method was applicable for determination of MMC in serum, although solid-phase extraction for sample clean-up was required prior to injection into the HPLC column, and analytical columns of 150-250 mm were necessary for adequate resolution of peaks. The method has been validated and is linear from 6.25 to 50 ng/ml in aqueous humor and from 10 to 500 ng/ml in serum. PMID- 8245156 TI - Simultaneous determination of flumazenil, midazolam and metabolites in human biological fluids by liquid chromatography. AB - A simple procedure for the simultaneous determination of flumazenil, midazolam, 1 hydroxymethylmidazolam and 4-hydroxymidazolam in plasma or urine in surgical patients is described. The assay involves a preliminary extraction of the drugs, metabolites and internal standard (flurazepam) from biological fluid into an organic solvent mixture (dichloromethane-diethyl ether, 40:60 v/v). The extract was evaporated to dryness at 40 degrees C under a gentle stream of nitrogen. The residue was re-dissolved in distilled methanol (80 microliters) and a 30 microliters aliquot was injected via an automatic sampler into the liquid chromatograph and eluted with the mobile phase (32% acetonitrile in 0.004 M sodium hydrogenphosphate buffer containing 1 ml of triethylamine and adjusted to pH 7.2) at a flow-rate of 1.5 ml/min on a 30-microns C8 precolumn linked to a 4 microns Nova-pak C18 cartridge column (100 mm x 8 mm I.D.) at ambient temperature (25 degrees C). The eluate was detected at 220 nm. PMID- 8245157 TI - Improved column-switching liquid chromatographic method for the determination of the enantiomers of mefloquine. AB - A liquid chromatographic method for the determination of the enantiomers of mefloquine has been improved. The chromatography involved two columns: an achiral cyanopropyl stationary phase for the quantification of (+/-)-mefloquine and a chiral naphthyl-urea stationary phase for the determination of the enantiomeric ratio. Compared with the previous method, which needed two detectors, this one used one detector-integrator to which the two columns are connected alternately by an automated column-switching system. The method is suitable for the quantification (0.05 microgram/ml) of mefloquine and the determination of enantiomeric ratios from 500-microliters plasma samples with ultraviolet detection. PMID- 8245158 TI - High-performance liquid chromatographic assay for N2-[5-(hypoxanthin-9 yl)pentyloxycarbonyl]-L-arginine (ST 789) in plasma by cyclization with benzoin and fluorimetric detection. AB - This paper describes a new highly sensitive assay for N2-[5-(hypoxanthin-9 yl)pentyloxycarbonyl]-L-arginine, an immunomodulatory agent, required for clinical pharmacokinetic investigation. A pre-column derivatization by cyclization with benzoin in aqueous medium produces the fluorescent 2-substituted amino-4,5-diphenylimidazole fluorescing at 450 nm (excitation wavelength 310 nm). L-Arginine-acetyl-L-carnitinamide chloride (ST 857, II), another arginine derivative, was used as an internal standard. A C18 DB column (5 microns, 250 mm x 4.6 mm I.D.) and a 45:55 (v/v) mixture of 0.05 M ammonium phosphate at pH 7.2 and methanol as mobile phase were used. Linearity was ascertained in the range 5 100 ng. Extraction recovery from plasma proved to be higher than 90% in the range 5-50 ng/ml. Intra-day precision, expressed as coefficient of variation, was in the range 4.7-6.0%. The limit of quantification proved to be 5 ng/ml and the limit of detection 2 ng/ml at a signal-to-noise ratio of 5. The method is specific. PMID- 8245159 TI - High-performance liquid chromatographic determination of concentrations of the reversible H+/K+ ATPase inhibitor SK&F 97574 in plasma. AB - A reversed-phase high-performance liquid chromatographic (HPLC) assay using ultraviolet spectrophotometric detection has been developed for the determination of the concentration of 3-butyryl-4-(2-methylphenylamino)-8-(2 hydroxyethoxy)quinoline (I) in rat, dog and human plasma. Prior to analysis, the protein in plasma samples was precipitated with acetonitrile containing 3-butyryl 4-(2-methylphenylamino)-8-methoxyquinoline to act as an internal standard. The supernatant layer was injected onto the HPLC column with no further clean-up. The assay requires 200 microliters of plasma and is precise and accurate within the range 25-1000 ng/ml. The mean within-run and between-run coefficients of variation were < 6% at 25 ng/ml and greater concentrations. The mean accuracy of quality control standards was generally within +/- 5% of the nominal concentration. Recovery of I and internal standard from plasma was approximately 100% over the entire assay range irrespective of species. PMID- 8245160 TI - Combined enzymic and chromatographic techniques to determine specific radioactivity in free and triglyceride fatty acid plasma fractions. AB - A reliable method to determine both free and triglyceride fatty acids and simultaneously to determine the specific radioactivity in each fraction has been developed. The procedure can be used to analyse a large number of samples. Lipoprotein lipase was used to hydrolyse triglyceride fatty acids, and a Carbopack B column was used to isolate free fatty acids. The radiolabeled fatty acids were determined by liquid scintillation counting, and individual fatty acid levels in each fraction were determined by gas chromatography. Free and triglyceride fatty acids were eluted in different fractions from the Carbopack B column. No interferences from other compounds were significant. PMID- 8245161 TI - Improved high-performance liquid chromatographic analysis of intracellular deoxyribonucleoside triphosphate levels. AB - The ability to measure intracellular deoxyribonucleoside triphosphate (dNTP) pool sizes is important for understanding the intracellular metabolism of DNA synthesis and repair. We have developed an improved method for measuring intracellular dNTP pool size by high-performance liquid chromatography (HPLC). Previous methods have enabled accurate measurement of dNTPs only in concentrations greater than approximately 10 pmol per 10(6) cells due to the inability to partially purify cell extracts, to the inability to apply extracts from extremely large numbers of cells, to the lack of efficient columns, to the presence of incompatible solvents, and to the inability to inject large volumes. We have modified a low-pressure strong anion-exchange column pre-step developed by others to concentrate and partially purify oxidized cell extracts while at the same time eluting them in a more compatible solvent for HPLC injection. The HPLC column is a YMC ODS-AQ column operating in a combined hydrophobic-interaction chromatography-reversed-phase chromatography mode. The injection and elution solvents are both phosphate-based. Using this method it is possible to measure intracellular dNTP levels well below 0.5 pmol per 10(6) cells or at the sensitivity of the DNA polymerase assay. PMID- 8245162 TI - Liquid chromatographic method for the determination of carotenoids, retinoids and tocopherols in human serum and in food. AB - A liquid chromatographic (LC) method has been developed for the quantitative measurement of the six major carotenoids in human serum (lutein, zeaxanthin, beta cryptoxanthin, lycopene, alpha-carotene, and beta-carotene) as well as retinol, retinyl palmitate, alpha-tocopherol, gamma-tocopherol, and delta-tocopherol. Several polar carotenoids, 2',3'-anhydrolutein, alpha-cryptoxanthin, and geometric isomers of lycopene and beta-carotene are also separated. Retinoids and carotenoids are monitored using a programmable ultraviolet-visible detector, while tocopherols are monitored using a fluorescence detector. The method uses a gradient containing acetonitrile, methanol, and ethyl acetate. Ammonium acetate is introduced with the methanol to minimize carotenoid losses on the LC column aggravated by the use of acetonitrile and ethyl acetate. The method is also applicable to the analysis of foods. PMID- 8245163 TI - Gas chromatographic-mass spectrometric method for determination of anabasine, anatabine and other tobacco alkaloids in urine of smokers and smokeless tobacco users. AB - A selected ion monitoring method for determination of the tobacco alkaloids anabasine, anatabine, nornicotine, metanicotine, dihydrometanicotine, and 2,3' bipyridyl in urine of smokers and smokeless tobacco users is described. The method involves conversion of the secondary amine alkaloids to tertiary amine derivatives by reductive alkylation using an aldehyde and sodium borohydride, and chromatography on a 5% phenylmethylsilicone capillary column. These derivatives have good chromatographic properties, allowing determination of concentrations as low as 1 ng/ml. The alkaloid 2,3'-bipyridyl is unaffected by the derivatization procedure and may be determined simultaneously with the other alkaloids. The structural analogues 2-(3-pyridyl)hexahydroazepine, 5-methyldihydrometanicotine, and 6-methyl-2,3'-bipyridyl were synthesized for use as internal standards. Using the method, concentrations and 24 h excretion of anabasine, anatabine, and nornicotine in urine of twenty-two smokers, eight chewing tobacco users, and six oral snuff users were determined and compared with concentrations and excretion of nicotine and its metabolite cotinine. Excretion of nicotine and cotinine was similar in all tobacco users, but excretion of anabasine, anatabine and nornicotine was substantially greater in urine of smokeless tobacco users, presumably due to absence of pyrolysis of these alkaloids in smokeless tobacco products. PMID- 8245164 TI - Determination of erythromycin A by liquid chromatography and electrochemical detection, with application to salmon tissue. AB - The chromatographic performance of erythromycin A (EA) is improved significantly over that achieved on polymeric columns by using a sterically shielded octyldiisopropylsilica (Zorbax Rx-C8) column and a neutral mobile phase consisting of 5 mM aqueous sodium perchlorate-acetonitrile (50:50). This mobile phase facilitates electrochemical detection of EA at the 3-pmol level. Temperature control of both column and detection cell is important for minimizing detector noise and drift. A clean-up procedure, based on aminopropylsilica solid phase extraction, allows the detection of EA in salmon flesh down to the 0.2-ppm level. Some of the metabolites of EA that retain the tertiary amine may also be detected by this method. PMID- 8245165 TI - Liquid chromatographic determination of para-substituted N,N-dialkylaniline N oxides. AB - A high-performance liquid chromatographic method for the determination of p substituted N,N-dialkylaniline N-oxides [N,N-dimethylaniline N-oxide (DMANO), N,N dimethyl-p-toluidine N-oxide, N,N-diethylaniline N-oxide, N-ethyl-N-methylaniline N-oxide (EMANO), p-cyano-N,N-dimethylaniline N-oxide (pCNDMANO), and N phenylpyrrolidine N-oxide (PPNO)] has been developed. It uses an octadecylsilica column, a mobile phase of methanol-phosphate buffer (pH 7.0, adjusted by triethylamine) and ultraviolet detection. N-Oxides were eluted in the order of hydrophilicity except for PPNO, which eluted between DMANO and EMANO. The number of theoretical plates and the detection limit under optimized conditions were between 2400 (DMANO) and 5400 (pCNDMANO) and between 2.3 microM (pCNDMANO) and 30 microM (PPNO), respectively. The concentration of N-oxides recovered from enzyme reaction mixtures by solid-phase extraction using Sep-Pak C18 (prior to chromatography) was also optimized with regard to the sensitivity and interference. PMID- 8245166 TI - Sensitive high-performance liquid chromatographic method for direct separation of labetalol stereoisomers in biological fluids using an alpha 1-acid glycoprotein stationary phase. AB - A chiral high-performance liquid chromatographic assay for the separation of the four stereoisomers of labetalol, an antihypertensive, in biological fluids has been developed. Baseline separation of the isomers was achieved using an alpha 1 acid glycoprotein stationary phase. No interference from endogenous substances was observed following extraction from various biological fluids obtained from pregnant (ewe and fetus) and non-pregnant sheep. The concentration of the individual isomers of labetalol was determined by first measuring the total concentration of racemic labetalol obtained from an achiral assay followed by reassay of each sample by the chiral method after which, by using the estimate of the percentage of each individual isomer, the individual concentration of each of the four isomers was determined. The mobile phase was 0.02 M phosphate buffer containing 0.015 M tetrabutylammonium phosphate. The pH of the mobile phase was adjusted to 7.10. The detector was set at an excitation wavelength of 230 nm and emission wavelength of 400 nm to monitor the nascent fluorescence intensity of the isomers of labetalol. The limit of detection of the individual isomers was 0.15 ng (0.6 ng of injected racemic labetalol). The assay was linear over the range 0.6-15.0 ng of labetalol (injected) with the intra- and inter-day mean coefficients of variation being less than 9.0 and 6.0%, respectively. Application of the assay in the study of pharmacokinetics of the stereoisomers of labetalol in sheep following administration of racemic labetalol has been demonstrated. PMID- 8245167 TI - Direct enantiomeric separation of cis-(+/-)diltiazem in plasma by high performance liquid chromatography with ovomucoid column. AB - A method is proposed for the direct enantiomeric separation of cis-(+/-)diltiazem in plasma. The method is based on a commercial Ultron ES-OVM column with a mobile phase of different mixtures of ethanol-phosphate buffer. The degree of extraction of the enantiomers was more than 86%. The detection limit of the method used was 5 ng of racemic mixture on column (coefficient of variation = 13%), corresponding to 3.2 ng for cis-(+/-)diltiazem and 1.8 ng for cis-(+/-)diltiazem. PMID- 8245168 TI - Method for isolation of non-esterified fatty acids and several other classes of plasma lipids by column chromatography on silica gel. AB - A method is described for isolation from human plasma of non-esterified fatty acids, cholesteryl esters, triglycerides, cholesterol and diglycerides, monoglycerides, and some phospholipids by extraction and silica gel column chromatography. All of these lipid classes except diglycerides and cholesterol were separated cleanly in seven elution steps. Diglycerides and cholesterol were isolated together. Recovery of model compounds which represent the most significant classes of plasma lipids during the column chromatographic step was nearly complete. The overall recovery of added heptadecanoic acid from plasma specimens was 81% after both sample isolation steps. The overall recovery of added synthetic pentadecanoic acid and heptadecanoic acid ester lipid homologues from plasma was 80-91% after both sample preparation steps. About 6 h are required for extraction and isolation in duplicate of these lipid classes from twenty plasma specimens. Alternatively, non-esterified fatty acids can be isolated from twenty plasma specimens in duplicate within 4 h by a variation of the full procedure. PMID- 8245169 TI - Direct determination of adamantanamine in plasma and urine with automated solid phase derivatization. AB - A simple, highly sensitive and selective method is described for adamantanamine determination in plasma and urine by high-performance liquid chromatography with fluorescence detection. The method involved a simultaneous extraction and derivatization of biological fluids with a 9-fluoreneacetate (9-FA) solid-phase derivatization reagent. This approach eliminated tedious sample preparation steps and provided automatic derivatization with selective and efficient sample clean up for direct injection of biological fluids. Derivatized adamantanamine was separated under conventional reversed-phase conditions and determined by fluorescence detection. The optimization and validation of the derivatization method with the 9-FA solid-phase reagent is described. PMID- 8245170 TI - Identification and determination of individual sophorolipids in fermentation products by gradient elution high-performance liquid chromatography with evaporative light-scattering detection. AB - High-performance liquid chromatography (HPLC) was used for the characterization of sophorolipids, one of the most important types of glycolipid biosurfactants. By using gradient elution with a water-acetonitrile mixture on a reversed-phase (C18) column and evaporative light-scattering detection, resolution of all the important individual sophorolipids present in fermentation products was achieved. In addition to HPLC, a combination of techniques involving selective production by fermentation of sophorolipids, chemical conversions of the products, separation methods and, for identification of lipidic chains of sophorolipids, gas chromatography and mass spectrometry was used. This led to the identification of almost all significant compounds observed in HPLC, including several previously unreported sophorolipids. As a result, a rapid method is now available for investigations of the influence of fermentation conditions on the nature and quantitative distribution of the sophorolipid products obtained. PMID- 8245171 TI - High-performance liquid chromatography of the photoproducts of nucleic acid components. III. Detection of the secondary structure differences in sequence isomeric self-complementary oligonucleotides. AB - Reversed-phase high-performance liquid chromatography (RP-HPLC) was used to detect differences in the secondary structures of two self-complementary oligodeoxyribonucleotides. The [d(CCTTTAAAGG)]2 duplex assumes an ordinary B conformation in aqueous solution, while [d(GGAAATTTCC)]2 is known to contain in its central part a stretch of a more rigid B'-form conformation with significantly lowered fluctuational mobility of base pairs. The latter factor causes a marked difference in the amounts of thymine cyclobutane photodimers formed under UV irradiation of corresponding duplexes as revealed by chromatography of two single-stranded oligonucleotides. Increasing the temperature below the duplex melting temperature (stimulation of the B'-B structural transition) results in an increase in photodimer formation that was inhibited in the B'-form. Thus, we demonstrate the usefulness of RP-HPLC for duplex DNA structural studies. PMID- 8245172 TI - Application of solid-phase microextraction to the headspace gas chromatographic analysis of halogenated volatiles in selected foods. AB - Solid-phase microextraction (SPME), with the poly(dimethylsiloxane)-coated silica fiber suspended and equilibrated in the headspace, has been applied to the capillary gas chromatographic (GC) analysis of 33 halogenated volatile contaminants in model aqueous solutions and in foods. With electrolytic conductivity detection, the limits of detection in water ranged from 1.5 micrograms/kg for vinyl chloride to < or = 0.005 microgram/kg for the tri- to hexachlorobenzenes. Headspace SPME-GC shows a much greater response for the less volatile analytes than those of greater volatility, a procedure complementing headspace GC with gas sampling. In model systems or foods, increasing lipid material decreased the headspace extraction. With 50 mg of lipid, the headspace extraction decreased about 50% for analytes with LODs about 0.1 microgram/kg and by > or = 99.5% for the above chlorobenzenes. Standard addition was used to analyze a variety of beverages and dry foods and to determine the analyte partitions. PMID- 8245173 TI - Separation of neuropeptide Y diastereomers by high-performance liquid chromatography and capillary zone electrophoresis. AB - Separation of analogues of neuropeptide Y (NPY) in which a single D-amino acid replaced the corresponding naturally occurring residue was performed by chromatographic techniques to ensure the quality of the synthetic peptides to be used for structural and biological studies. Of the 35 compounds, 28 were easily separated (alpha = 1.02-2.76) from native NPY by standard reversed-phase high performance chromatography (RP-HPLC) methods using a Vydac C18 column and a gradient buffer system developed in our laboratory comprised of triethylammonium phosphate (TEAP) at pH 2.25 and acetonitrile at 40 degrees C. The identical diastereomers could be separated on the same solid support and by using 0.1% trifluoroacetic acid (TFA) as the mobile phase modifier, however separation factors were smaller and retention times were longer. Three of the remaining seven unresolved analogues were separated (alpha = 1.02-1.96) by changing the solid-phase support to Vydac diphenyl derivatized silica and a buffer system consisting of 0.1% TFA and acetonitrile. Of the four remaining unresolved analogues, only two could be separated by capillary zone electrophoresis (CZE) in 0.1 M sodium phosphate at pH 2.5, but all four were finally resolved by changing the electrophoretic buffer to 0.1 M TEAP buffer at pH 2.5. Migration times of the diastereomers differed by 0.2-2.0 min from that of the natural NPY. In addition to confirming the uniqueness of each isomer, this investigation demonstrated the expansive utility and high efficiency of the TEAP buffer system for both RP-HPLC and CZE as well as the difference in selectivity produced by the TEAP and TFA buffers in RP-HPLC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245174 TI - Comparison of a liquid solvent extraction technique and supercritical fluid extraction for the determination of alpha- and beta-carotene in vegetables. AB - An ethanol-pentane solvent extraction procedure and a supercritical CO2 extraction procedure are compared for the high-performance liquid chromatographic determination of alpha- and beta-carotene in vegetables. The vegetables tested included carrots, collard greens, turnips, turnip greens, kale, mustard greens, broccoli florets, zucchini, and squash. Homogenization of the sample prior to liquid or supercritical fluid extraction significantly improved recovery of the carotenoids. A combination of static and dynamic modes of extraction with ethanol modifier at 338 atm and 40 degrees C was necessary in order to achieve optimum recovery with the supercritical fluid procedure. beta-Carotene results with the supercritical CO2 procedure averaged 23% higher than results for the liquid extraction procedure. Only corn and carrots contained detectable levels of alpha carotene, and, in both cases, liquid extraction yielded slightly higher results. Liquid extractions were performed in approximately 90 min, and supercritical fluid extractions were performed in 30 min; however, the supercritical fluid extractions procedure required less than 10 min of an analyst's time while the liquid extraction procedure was labor intensive. PMID- 8245175 TI - Determination of anabolic steroid 19-nor-testosterone in bovine serum by GC-SIM MS. AB - Anabolic steroids extracted from bovine serum by means of organic solvents or solid-phase extraction techniques with C18 cartridges are not suitable for quantitative analysis by gas chromatography-mass spectrometry (GC-MS); therefore, they were extracted from serum by Extrelut columns (Merck; Germany), eluted with diethyl ether, and purified by a solid-phase extraction technique using amino (NH2) columns. The resulting clean extract was treated with heptafluorobutyric anhydride (HFBA); this formed a derivative suitable for GC-MS analysis. Using selected ion monitoring (SIM), quantities as small as 40 pg/mL can be detected. PMID- 8245176 TI - Intravenous immunoglobulin induces interferon-gamma and interleukin-6 in vivo. AB - Immunoglobulin is known to be an immunomodulator. It can induce protein mediators from mononuclear cells, particularly monocytes in vitro. Intravenous immunoglobulin (IVIg) has been used as a therapy in several clinical situations. In this study, the influence of IVIg infusion on the plasma levels of two protein mediators, interferon-gamma (IFN-gamma) and interleukin-6 (IL-6), was assessed in patients with secondary generalized epilepsy. Compared to preinfusion levels, plasma interferon-gamma was increased in 18 of 18 patients 20 min after the 6- to 8-hr infusion of IVIg. Plasma interferon-gamma levels reached their peak at various times from 20 min to 3 days post IVIg infusion, dependent upon the individual patient. Plasma IL-6 levels also increased after IVIg infusion. Generally, IL-6 reached its peak level after IFN-gamma. No activated T cells or B cells were observed as determined by the expression of surface CD25, CD23, and HLA-DR 20 min following the infusion when the IFN-gamma and IL-6 levels were assessed. The expression of the high-affinity receptor for IgG, CD64, on monocytes was significantly enhanced after IVIg infusion, while the low-affinity receptor for IgG, CD32, was only slightly increased. Cytoplasmic staining of PBMC indicates that both CD16-positive and CD16-negative cells may contribute to the increase seen in plasma IFN-gamma. These data raise the possibility that the therapeutic effects of intravenous immunoglobulin may be related, at least in part, to the immunomodulatory activity as demonstrated by the changes in plasma levels of IFN-gamma and IL-6. PMID- 8245177 TI - Regulation of interleukin-6 (IL-6) expression: evidence for a tissue-specific role of protein kinase C. AB - The regulation of IL-6 mRNA expression was studied in human blood monocytes and in the human epidermoid carcinoma cell line HEp-2. In human monocytes phorbol-12 myristate 13-acetate (PMA) did not induce IL-6 but it increased IL-1 beta and IL 8 mRNA levels. Furthermore, in monocytes, protein kinase C (PKC) activation by PMA even reduced IL-1-induced IL-6 mRNA, and IL-1-induced IL-6 synthesis was increased by the PKC inhibitor staurosporine. IL-6 synthesis in HEp-2 cells was induced by IL-1, PMA, and calcium ionophore A 23187 but not by dibutyryl-cAMP. PMA-, but not IL-1-induced IL-6 synthesis in HEp-2 cells was inhibited by staurosporine. PMA pretreatment of HEp-2 cells abolished PMA-induced IL-6 but the IL-1 effect was not reduced. These data indicate that IL-6 can be induced by a PKC-independent pathway in monocytes and HEp-2 cells. In monocytes PKC activation does not induce IL-6 and PMA interferes with the IL-1 effect. Transcription factors known to be involved with the regulation of IL-6 expression were studied by gel retardation assays. NF-IL-6 and AP-1 activity were constitutively expressed in monocytes and HEp-2 cells under conditions where IL-6 mRNA was not detectable and levels did not change in response to stimulation by IL-1 or PMA. In contrast, NF-kB was increased by both IL-1 and PMA, but only the effect of PMA, and not that of IL-1, was inhibited by staurosporine. In summary, these results show tissue-specific differences in the regulation of IL-6 expression. Induction of IL-6 in monocytes is PKC independent. In the epithelial cell line HEp-2 IL-6 is inducible by PKC as well as by a PKC-independent pathway. PMID- 8245178 TI - Evaluation of soluble tumor necrosis factor (TNF) receptors and TNF receptor antibodies in patients with systemic lupus erythematodes, progressive systemic sclerosis, and mixed connective tissue disease. AB - Two TNF binding proteins have been characterized as soluble fragments of TNF receptors. We measured the plasma concentrations of soluble type A (p75) and type B (p55) TNF receptors in patients with systemic lupus erythematodes (SLE), progressive systemic sclerosis (PSS), and mixed connective tissue disease (MCTD). In SLE and PSS patients plasma concentrations of both types of TNF receptors and in MCTD patients type A TNF receptors were significantly elevated compared to controls. Plasma concentrations of both soluble TNF receptors were highly correlated in SLE, PSS, and MCTD patients, indicating a possible coregulation of both TNF receptors. In contrast, soluble interleukin 2 receptor (sCD 25) plasma concentrations were not correlated and seem to be an independent parameter. The soluble forms of the TNF receptors neutralize TNF in cytotoxicity assays and are functionally active as TNF antagonists. In one patient with SLE, autoantibodies against type A TNF receptors were detected, TNF alpha, and TNF beta did not interfere with the autoantibody binding to the receptor. PMID- 8245179 TI - Anti-"dividing cell antigen" autoantibody: a novel antinuclear antibody pattern related to histones in systemic lupus erythematosus. AB - This study aims to describe a novel antinuclear antibody directed to proteins only accessible during the mitosis: anti-"dividing cell antigen" (DCA) antibody. A total of 709 disease-associated and control sera was tested by indirect immunofluorescence using a variety of cell lines as substrate. Cells were treated with enzymes and antibodies absorbed with nuclear antigens. Antibodies to DNA, histone subfractions, and synthetic peptides were evaluated using enzyme-linked immunosorbent assays. Cell extracts were electrophoresed before and after synchronization and sera tested on the blots. The anti-DCA antibody was demonstrated in 10 of 183 SLE patients but virtually never in other connective tissue diseases. The DCA was sensitive to HCl and proteolytic enzymes and the anti-DCA binding inhibited by histones H2A and H2B. Differences of anti-H2A and anti-H2B were observed between anti-DCA antibody-positive and anti-DCA antibody negative sera, and antibodies specific for the 1-15 region of H2A, the 1-25 region of H2B and the 1-29 region of H4 were more frequent in the former sera than in the latter. The anti-DCA antibody was shown to react with a 60-kDa protein. Our findings suggest that the anti-DCA antibody is directed to a protein complex containing H2A and H2B. PMID- 8245180 TI - Catecholamines induce alterations of distribution and activity of human natural killer (NK) cells. AB - Catecholamines have been suggested to be responsible for altered cellular immunity after stress. This study was performed to determine the effects of adrenaline and noradrenaline on lymphocyte subpopulations and NK cell functions. Subjects were given a subcutaneous injection of either NaCl, adrenaline (5 micrograms/kg), or noradrenaline (10 micrograms/kg). Catecholamine concentrations, subsets of peripheral blood lymphocytes, NK activity, and antibody-dependent cellular cytotoxicity (ADCC) were analyzed before (baseline) and 5, 15, 30, 60, and 120 min after injection. There were no differences between groups in the distribution of CD2+ and CD8+ lymphocytes over time. However, CD3+ and CD4+ T cells decreased significantly 5 to 60 min after injection of adrenaline. In contrast, NK cell numbers (CD16+, CD56+) increased significantly 5 min after injection of adrenaline and noradrenaline, reached the highest values 15 to 30 min postinjection, and subsequently declined to baseline values 60 (noradrenaline) and 120 (adrenaline) min, respectively, after injection. Similar alterations for NK activity and ADCC were observed after administration of both catecholamines. These data suggest that both sympathetic-adrenal hormones are similarly potent modulators of natural immunity and provide further evidence that catecholamines might be responsible for the observed alterations in immune functions after phases of acute stress. PMID- 8245181 TI - Cellular immune response of patients with uveitis to peptide M, a retinal S antigen fragment. AB - Peptide M, an 18-amino acid fragment from position 303 to position 320 of retinal S-antigen, produces experimental autoimmune uveitis (EAU), similar to that produced by native S-antigen, in several vertebrate species including nonhuman primates. It was observed that 12 of the 39 (30.7%) patients with uveitis, 1 of the 29 (3.4%) patients with systemic connective tissue disorders (CTD) without eye involvement, 2 of the 7 (5.8%) patients of CTD with uveitis, 1 of the 17 (5.8%) patients with diabetic retinopathy, and none of the 19 normal healthy controls showed a significant lymphoproliferative response to peptide M (stimulation index of 3 or more). Yeast histone H3 peptide gave a positive response in 1 (2.5%), 2 (6.8%), 1 (14.2%), 2 (11.7%), and 2 (10.5%) individuals, respectively, in the different groups studied. In a few cases a positive response to yeast histone H3 peptide was observed without significant stimulation to peptide M. These findings indicate that peptide M could also be an immunogenic epitope of S-antigen in humans and be aetiopathologically related to uveitis in a subset of patients with this disease. However, unlike experimental animals, the responses to peptide M and yeast histone H3 were nonconcordant, necessitating further studies. PMID- 8245182 TI - Anti-Pseudomonas aeruginosa IgG subclass titers in patients with cystic fibrosis: correlations with pulmonary function, neutrophil chemotaxis, and phagocytosis. AB - To explore possible mechanisms for the association between elevated immunoglobulin levels and lower pulmonary function in cystic fibrosis patients, we measured serum IgG subclass levels and anti-P. aeruginosa IgG subclass titers and correlated levels with neutrophil phagocytosis and chemotaxis. Serum was obtained from 13 cystic fibrosis patients colonized with the same serotype of P. aeruginosa, 12 noncolonized patients, and 12 normal volunteers. All anti-P. aeruginosa IgG subclass titers were elevated in serum from colonized patients. IgG3 level and anti-P. aeruginosa IgG3 titer were inversely correlated with pulmonary function. Phagocytosis of P. aeruginosa by neutrophils correlated with serum IgG3 level and was increased by opsonization with serum from colonized patients. Chemotactic index was increased in serum from colonized patients and inversely correlated with pulmonary function chest roentgenogram score. Chemotactic index directly correlated with anti-P. aeruginosa IgG3 titer and serum IgG3. These data demonstrate that cystic fibrosis patients with increased IgG3 levels are in poorer clinical condition and that their serum enhances neutrophil function. Such patients may have increased pulmonary inflammation with subsequent lung damage. PMID- 8245183 TI - A farewell to symmetry. PMID- 8245184 TI - 'Animated' orthodontic treatment planning. PMID- 8245185 TI - Selection card for ligature colors. PMID- 8245186 TI - An embedded lingual holding arch. PMID- 8245187 TI - Implant therapy in cases of agenesis. PMID- 8245188 TI - Self-directed orthodontic teams. PMID- 8245189 TI - Informed consent for credit checks. PMID- 8245190 TI - Late knee problems in myelomeningocele. AB - This study was undertaken to assess the incidence and etiology of knee problems in a long-term follow-up of myelomeningocele patients. Of the 72 community ambulators reviewed, 17 (24%) had significant knee symptoms. A specific gait pattern was identified in symptomatic patients with low lumbar lesions, which may be explained on the basis of hip abductor and calf muscle weakness. These patients have a characteristic gait, which places abnormal stress on the knee, leading to medial and anteromedial rotary instability and eventual degenerative change. It is likely that disability resulting from knee symptoms will be the factor precluding independent ambulation in the patients reviewed in this study. PMID- 8245191 TI - Comparison of two instrumentation techniques in treatment of lumbar kyphosis in myelodysplasia. AB - Thirty-three patients with myelodysplasia had kyphectomies for correction of spinal deformities. Harrington compression instrumentation was used for fixation in 21 patients, with postoperative immobilization in a spinal orthosis for 6 months. Twelve patients had Luque rod instrumentation, with fixation through the first sacral foramina with a modification of the Dunn technique and with no postoperative immobilization. Kyphosis recurred in eight patients with Harrington rod instrumentation and in none of those with Luque rod instrumentation. Nine patients with Harrington compression rods required 15 procedures for instrumentation problems. Only five of the 21 patients with Harrington compression rods had no significant complications, although ultimately 20 of 21 patients had solid fusions without further progression of the kyphosis. All 12 patients with Luque rods had solid fusions with no recurrences of the kyphotic deformity. One patient required operation for instrumentation problems. PMID- 8245192 TI - Latex allergy in spina bifida patients: prevalence and surgical implications. AB - We interviewed 93 consecutive children with spina bifida who were scheduled for operation to document (a) the prevalence of latex sensitivity, (b) predictors of anaphylactic reaction, and (c) the risk of type I hypersensitivity reaction perioperatively. Of these patients, 9.7% were clinically allergic. Radioallergosorbent testing (RAST) was 89% sensitive. All patients with known latex allergy were premedicated, and rigorous efforts were made to avoid latex exposure. No intraoperative anaphylactic reactions were noted. One postoperative reaction occurred in a history-positive, RAST-negative patient. We conclude that operation can be performed safely in patients with latex allergy. PMID- 8245193 TI - Long-term follow-up of triple arthrodesis in patients with cerebral palsy. AB - This study was a long-term retrospective review of patients with cerebral palsy (CP) who had triple arthrodeses as children before 1981. The medical records were reviewed, and the patients were requested to return for reevaluation, during which a radiograph, physical examination, and patient questionnaire were obtained. Twenty-four patients who had triple arthrodesis on 35 feet returned for evaluation. Twenty-three feet had planovalgus deformities and 12 equinovarus deformities. Mean age at operation was 14.2 years, with a mean follow-up of 17.8 years (range 11-45 years). Of the 24 patients questioned, 19 were satisfied and five were dissatisfied with their result. Nine patients had occasional pain; one patient reported frequent pain in one foot. Six patients had limited distance ambulation owing to their feet. Radiographic evaluation demonstrated that 43% of the feet had degenerative changes at the ankle joint. Four of six patients reported ambulatory limitation due to pain. Ankle joint range of motion (ROM) and degenerative arthritis were not correlated with pain, distance limitations, residual deformity, or patient satisfaction. Patient satisfaction was predominantly related to persistent pain, especially pain causing distance limitations in ambulation. Patient satisfaction was also strongly correlated with residual deformity. Persistent pain and distance limitation were also strongly correlated with residual planovalgus deformity. PMID- 8245194 TI - Significance of the confusion test in cerebral palsy. AB - The confusion test examines ankle dorsiflexion in patients with cerebral palsy. Orthopedists have related this test to swing-phase activity of the tibialis anterior, and have used it as a prerequisite for tendon transfer. To determine the validity of this assumption, ankle dorsiflexion was tested in 47 normal children. Forty-seven percent had a positive, unresisted confusion test, and 97% had a positive, resisted confusion test. Twenty-three patients with cerebral palsy who had a positive confusion test underwent gait analysis. Tibialis anterior electromyographs showed wide variability. Sagittal-plane ankle-movement curves revealed five patterns. Thirty-three percent of the patients showed abnormal swing-phase dorsiflexion, and 61% had abnormal swing-phase plantar flexion. We conclude that the confusion test evaluates a normal, patterned response, and is positive in most children with cerebral palsy. Although a positive confusion test shows that active ankle dorsiflexion is possible, it is not predictive of swing-phase ankle kinematics. PMID- 8245195 TI - Hamstrings in cerebral palsy crouch gait. AB - After observing patients with increased anterior pelvic tilt following medial hamstring lengthening in cerebral palsy crouch gait, we became concerned that the hamstrings may be functionally important hip extensors. To evaluate this, we studied the three-dimensional motion of the hip and knee, calculated hamstring muscle length, and evaluated dynamic electromyography (EMG) of the medial hamstrings in 16 patients with diplegic cerebral palsy and crouch gait to determine if the hamstrings were extending the hip. Twelve of 16 patients exhibited marked prolongation of electrical activity in the medial hamstrings, and in eight of these 12, the hamstrings were contracting concentrically, thus aiding in hip extension during gait. Hamstrings may be important hip extensors in some cerebral palsy patients with crouch gait; however, other deformities contributing to crouch (such as hip flexion contracture) need to be considered before isolated hamstring lengthening is performed in these patients. PMID- 8245196 TI - Kinematic and kinetic evaluation of the ankle after lengthening of the gastrocnemius fascia in children with cerebral palsy. AB - The effect of surgical lengthening of the gastrocnemius fascia on ankle joint kinematics and kinetics during gait in patients with cerebral palsy (CP) was evaluated. Twenty independent ambulators (24 sides) were included in this retrospective study. The evaluation included clinical examination, calculation of joint kinematics and kinetics, and collection of surface electromyography (EMG) during gait. Postoperative improvements were noted in static heelcord range of motion (ROM), with an associated increase in dorsiflexion in stance and swing. Kinetic analysis showed a decrease in the abnormal energy generated around the ankle in midstance and a statistically significant increase in the energy generated in late stance for push-off. PMID- 8245197 TI - Heterotopic ossification after rhizotomy and femoral osteotomy. AB - In 15 patients with cerebral palsy who underwent selective posterior rhizotomy (SPR) and subsequent proximal femoral varus derotation osteotomy (VDO), heterotopic ossification (HO) around the hip after VDO was noted in four of eight patients with spastic quadriplegia (seven of 26 hips, 27%). HO was not noted in the seven patients with spastic diplegia. A radiographic review of 118 hips with a femoral VDO in the 69 patients with cerebral palsy who did not undergo SPR during the same period showed no HO. Frequent evaluation of postrhizotomy patients with cerebral palsy quadriplegia after a femoral VDO is necessary to recognize HO as a possible etiology for late-onset pain and limited hip range of motion (ROM). PMID- 8245198 TI - Heterotopic ossification after hip and spine surgery in children with cerebral palsy. AB - We made a retrospective review of 198 children with cerebral spasticity to determine development of heterotopic ossification. Of 61 children undergoing hip adduction lengthening, 21 had mild to moderate grades of heterotopic ossification. Only two of 132 patients who underwent spine fusions had developed heterotopic ossification, but in both it was severe, resulting in hip fusion. Two of five patients undergoing concomitant hip surgery and spine fusion developed heterotopic ossification of severe degree. The occurrence of heterotopic ossification after hip muscle surgery in children with cerebral spasticity is common but usually minor. Heterotopic ossification developing after spine fusion is rare but severe when it occurs, and concomitant hip and spine surgery causes frequent severe occurrences of heterotopic ossification. PMID- 8245199 TI - Soft-tissue expander failure in severe equinovarus foot deformity. AB - Soft-tissue expanders have recently been reported to be useful in clubfoot surgery. We report our experience with this procedure in five patients with seven severe equinovarus foot deformities. The primary diagnoses of these patients included distal arthrogryposis, cerebro-oculo-facial syndrome, ischemic contracture and spina bifida. Our patients' major complications included ischemia, infection, and sepsis, which resulted in premature removal of the soft tissue expanders in five of seven clubfeet. Minor complications, such as partial wound dehiscence, occurred in the two successful expansions. Predisposing factors may include soft-tissue expanders with self-contained ports, poor compliance, and extremely tight tissues that limit the subcutaneous space available for expansion, with resultant diminution of the vascular supply to the overlying skin. PMID- 8245200 TI - Simplified technique for scapulocostal fusion in facioscapulohumeral dystrophy. AB - Surgical stabilization of the scapula by a simplified technique is suggested. An average gain of nearly 30 degrees in both shoulder abduction and flexion with elimination of scapular winging resulted. In all cases, this allowed elevation of the arm above the head, with concomitant improvement in activities of daily living (ADL). There were no instances of pleuritic chest pain, hemothorax, or pneumothorax. There was no substantive change in pulmonary function. Scapulocostal fusion in patients with facioscapulohumeral (FSH) dystrophy is an effective operation to improve upper extremity function. Patients report a subjective increase in endurance and are uniformly pleased with the cosmetic result. PMID- 8245201 TI - Spinal fusion in Duchenne muscular dystrophy--fixation and fusion to the sacropelvis? AB - Twenty-two wheelchair-bound patients with Duchenne muscular dystrophy (DMD) underwent Luque segmental instrumentation and fusion. Twelve patients were instrumented to the sacropelvis, and 10 were instrumented to L5. Mean preoperative and postoperative curves were nearly identical in both groups. The mild degree of trunk shift and pelvic obliquity was similar between the two groups. The recommendation for operation in such patients should be made when their curve is > 20 degrees and if their forced vital capacity is > 40%. If treatment is initiated early, Luque instrumentation and fusion from high thoracic (T2 or T3) to the fifth lumbar vertebra should be sufficient. PMID- 8245202 TI - Normal values of neonatal lower limbs: an evaluation of 1,000 neonates. AB - The purpose of this study was to establish normal values for neonatal hip range of motion, knee range of motion, tibial torsion, and foot alignment. Normal values for examination of the lower extremity in the neonate are not well documented in the orthopaedic literature. One-thousand healthy neonates were examined by the orthopaedic residency under the supervision of an attending doctor. A total of 527 girls (53%) and 473 boys (47%) were examined. Results were compared and analyzed for mean, range, and standard deviations. There was a wide normal range for each category measured. For all categories, boys and girls had essentially the same values. There were no differences between right and left values. Normal values for these measurements have not previously been reported from such a large series. PMID- 8245203 TI - Predicting height from arm measurements. AB - Height measurement is required to standardize measures of physical capacity (e.g., pulmonary function) and to adjust drug dosage in the physically disabled. To estimate height in wheelchair-bound patients, we examined the relation of four different upper extremity measurements to height in 119 normal individuals aged 0.5-56 years. Regression analysis was performed for each of the following: (a) interacromioclavicular distance, (b) upper arm, (c) lower arm (cubit), (d) arm span; with height. Pearson correlation coefficients of 0.937, 0.967, 0.975, and 0.989, respectively, were obtained (accuracy +/- 5%, 90% confidence interval, for upper and lower arm measurements). We conclude that height can be predicted reliably from arm measurements. PMID- 8245204 TI - Ultrasound localization of subperiosteal abscesses in children with late-acute osteomyelitis. AB - Ultrasonography was undertaken in nine children with late presenting acute osteomyelitis, in four children with typical superficial cellulitis, and in four with a soft tissue abscess. Ultrasound distinguished between superficial cellulitis, soft tissue abscess, and subperiosteal abscess. The abscesses were confirmed at operation, and a subperiosteal abscess was also detected in the child with deep periosseous cellulitis. Ultrasonography was particularly useful in confirming the existence of a subperiosteal abscess and in localizing it precisely in children with diffuse swelling and tenderness of a limb owing to late-acute osteomyelitis. Surgical drainage of pus can be avoided in patients without ultrasound features of an abscess and can be better planned in those who require it. PMID- 8245205 TI - Pyomyositis in children. AB - Bacterial infections of muscle, also known as pyomyositis or tropical pyomyositis, occur more commonly in tropical regions. This article reviews five cases of pyomyositis diagnosed in children over a 1-year period. Pyomyositis should be considered in the differential diagnosis of septic-appearing children as well as children complaining of joint pain or muscle aches. The diagnosis can be aided with either a computed tomography (CT) or magnetic resonance imaging (MRI) scan. The MRI is preferable because multiple processes can be evaluated, such as joint effusion suggesting septic arthritis. If the patient does not respond quickly to antibiotics and surgical intervention, either there is a recurrence of the previously debrided abscess, or there is an unrecognized secondary abscess. Multiple abscess sites should be entertained prior to initial debridement. PMID- 8245206 TI - Hypertension following orthopaedic surgery in children. AB - Five patients developed hypertension after undergoing orthopaedic surgery. Three of four patients in whom serum renin was assayed had significant elevation, suggesting a possible renal-mediated etiology. Two patients required the addition of an angiotensin-converting enzyme (ACE) inhibitor to control hypertension while continuing the prolonged orthopaedic procedures. The need for careful blood pressure monitoring of children undergoing orthopaedic surgery is emphasized in this article. PMID- 8245207 TI - Intraoperative blood salvage in children and young adults undergoing spinal surgery with predeposited autologous blood: efficacy and cost effectiveness. AB - We conducted a retrospective review of 155 spinal operations at our institution to determine the efficacy of intraoperative salvage. Addition of intraoperative salvage had little effect on the success of a preoperative autologous donation program. Only patients with operative blood loss > 2,000 ml (12% of patients) benefited from this expensive source of autologous blood. The technique tended to be most effective in children aged 16-18 years. Use of intraoperative salvage for all pediatric spinal procedures is neither necessary nor cost effective. PMID- 8245208 TI - Erythrocyte viability in postoperative autotransfusion. AB - Awareness of the potential for transmission of the spread of viral diseases through homologous blood products has led to an increasing awareness of techniques that minimize these risks. Reinfusion of postoperatively shed blood has become popular, but its efficacy has been questioned. In an effort to determine the viability of postoperatively collected cells, 10 consecutive patients undergoing an identical surgical procedure were studied; their blood was tagged with chromium 51 and reinfused. Reinfused cells were then monitored serially at determined intervals and expressed as a percentage of postinfusion activity. At 4 days, 75.9% of initial activity remained. These figures are comparable to those previously reported for intraoperative and preoperative cell collection. Blood salvaged postoperatively is not significantly damaged and is as viable as blood collected from other autologous and homologous sources. PMID- 8245209 TI - Focal scleroderma in children: an orthopaedic perspective. AB - Focal scleroderma is a connective-tissue disorder manifested by fibrosis of the skin and subcutaneous tissues. Consequently, it may be associated with joint contractures, extremity deformity, and impairment of extremity function. It has a variable clinical course, with both remissions and recurrences. In resistant cases of extremity involvement, treatment considerations should include physical and occupational therapy, medical treatment, bracing, serial casting, and surgical intervention. We reviewed seven patients treated between 1960 and 1990 with significant joint contractures secondary to focal scleroderma. Serial casting was found to be useful for contractures about the wrist, knee, and ankle. Soft-tissue release was an effective treatment for contractures at the wrist, hip, and ankle. Knee disarticulation was performed on one patient with a severe knee-flexion contracture. Epiphysiodesis and femoral shortening osteotomy were effective treatments for leg-length equalization in a case of hemiatrophy. PMID- 8245210 TI - Retinal detachment in spondyloepiphyseal dysplasia congenita. AB - Retinal detachment is one of the most serious complications of spondyloepiphyseal dysplasia congenita (SEDC). The ocular complication of 12 patients with SEDC was studied. Three patients developed bilateral retinal detachments; five of the six detachments occurred during the period of rapid growth spurt. Our results suggest that patients with SEDC have an increased risk for development of retinal detachment during the period of rapid growth spurt. PMID- 8245211 TI - A case of Satoyoshi's syndrome. AB - Satoyoshi's syndrome is a rare neurologic disorder characterized by progressive muscle spasms, alopecia, diarrhea, and skeletal abnormalities. We describe a 21 year-old woman with Satoyoshi's syndrome whose skeletal changes were followed for 15 years until after she had reached skeletal maturity. The patient developed slipping of the epiphyses in the proximal humeri and femurs and metaphyseal lesions mimicking metaphyseal chondrodysplasia. These lesions were a consequence of physeal injuries caused by the recurrent muscle spasms. PMID- 8245212 TI - Hallux interphalangeus extensus. AB - We report four cases of symptomatic extension deformity of the interphalangeal joint of the hallux. Patients without pathology in the joint were effectively treated by osteotomy of the proximal phalanx. For older patients, or those with joint pathology, interphalangeal fusion is recommended. The condition has not been previously reported. PMID- 8245213 TI - A modification of the Cincinnati approach for obtaining adequate lengthening of the tendoachilles in the presence of severe equinus deformity. PMID- 8245214 TI - Proximal femoral osteotomy using the AO fixed-angle blade plate. PMID- 8245215 TI - Cytoarchitecture of spinal-projecting neurons in the brain of the larval sea lamprey. AB - The descending spinal projecting system of the lamprey is of interest because it includes axons that activate swimming pattern generators and because regeneration of this system is involved in the behavioral recovery of lampreys following spinal transection. However, little is known about the true size of this projection and of the distribution of its terminations along the spinal cord. Brain neurons with spinal projections were studied in larval sea lampreys by using wholemount preparations labeled retrogradely with horseradish peroxidase (HRP) from spinal injections at 10%, 15%, 25%, 50%, 70%, and 75% of body length from the anterior end. Neurons projecting to different levels of the spinal cord were mapped. A large number of descending axons terminated within nine segments caudal to the last gill. The spinal projection system was divided into 10 bilateral groups based on cytoarchitectural landmarks. All of the lateral nuclear groups had contralateral spinal projections. In addition to the 12 pairs of Muller cells, the pair of Mauthner cells, and the pair of auxiliary Mauthner cells described by previous authors, the study revealed four pairs of smaller neurons that were individually identifiable. PMID- 8245216 TI - Synaptic relationships involving local axon collaterals of pyramidal neurons in the cat motor cortex. AB - The intracortical synaptic relationships of pyramidal neurons in the cat motor cortex were studied by intracellular recording and labeling techniques. Neurons that responded with monosynaptic excitatory postsynaptic potentials (EPSPs) to microstimulation in the somatosensory cortex were identified by intracellular recordings. Long-term potentiation (LTP) was evoked in all of these neurons (n = 15), following tetanic stimulation (50 Hz, 5 s) of their afferents from the somatosensory cortex. Three of these cells (cells A-C) were identified as pyramidal neurons, following intracellular injections of Neurobiotin. The intracortical axon collaterals of these labeled cells arborized extensively, forming terminal clusters both in close proximity to the parent soma and along their long, horizontal branches. Terminal clusters in both the proximal and in the distal termination zones of each of the cells were studied by electron microscopy. In their proximal arborization zones, the axon collaterals of the labeled pyramidal neurons synapsed preferentially with dendritic spines belonging to other pyramidal cells. In contrast, in their distal terminal clusters, the axon collaterals of each of the cells formed synapses in different proportions with different postsynaptic targets. The distal axon collaterals of cell A formed 86% of their synapses with pyramidal neurons; those of cell B formed 64% of their synapses with pyramidal cells, the remaining synapses with the dendritic shafts and somata of nonpyramidal neurons, and those of cell C provided most of their output (68%) to nonpyramidal, presumably inhibitory neurons. These findings suggest a high selectivity of intrinsic axon collaterals to form specific patterns of synapses. The patterns of synaptic interactions formed by these intrinsic axon collaterals may be a substrate for shaping and modulating representation maps in the motor cortex. PMID- 8245217 TI - NGF augmentation rescues trigeminal ganglion and principalis neurons, but not brainstem or cortical whisker patterns, after infraorbital nerve injury at birth. AB - Prior studies indicate that neonatal nerve injury kills many trigeminal (V) first and second-order cells, and interrupts pattern formation in the brainstem and cerebral cortex. Yet it is not known whether effects upon cell survival and pattern formation are causally related. To determine whether axotomized V ganglion cells can be rescued by an exogenous trophic agent, rats received 5 mg/kg of nerve growth factor (NGF) prior to, and every day after, infraorbital nerve section on the day of birth until sacrifice on postnatal day (PND) 1, 3, 5, 7, or 14. Other animals received identical lesions without NGF. Ganglion cell numbers were significantly reduced by PND1 in pups not given NGF, while NGF treated rats displayed no significant cell loss through PND7. However, NGF did not permanently rescue V neurons because ganglion cell numbers were reliably reduced by PND14. Cell numbers in V nucleus principalis were reduced by PND1 in pups not given NGF, while NGF-treated animals displayed no cell loss through PND14. NGF's rescue of second-order cells is probably an indirect effect of NGF action upon V ganglion cells because, in other newborns, NGF failed to maintain principalis cells after direct lesion of the left V ganglion. To determine whether preventing cell death permits whisker-related pattern formation, other rats also received NGF prior to and after infraorbital nerve section at birth. After 3-14 days, patterns were assessed in the brainstem and cortex with cytochrome oxidase histochemistry and serotonin immunocytochemistry. Whisker related patterns failed to develop as in cases not given NGF. These data indicate that communication with the periphery is necessary for the maintenance of central whisker-related patterns. They also suggest that V ganglion cells can be rescued, albeit temporarily, from rapid injury-induced death by NGF, thereby delaying injury-induced cell death in nucleus principalis. However, the mechanism(s) responsible for injury-induced pattern alterations in the developing V system remains to be elucidated. PMID- 8245218 TI - The inhibitory motoneurons of crayfish thoracic limbs: identification, structures, and homology with insect common inhibitors. AB - The three inhibitory motoneurons supplying crayfish thoracic limbs were identified, stained, and compared structurally. The inhibitors to the walking leg muscles (in Orconectes) were identified anatomically by a combination of immunocytochemical staining for gamma-aminobutyric acid (GABA) or glutamate decarboxylase and differential backfill staining with nickel and cobalt ions. The cheliped inhibitors were identified intracellularly and injected with Lucifer Yellow (Pacifastacus) or cobalt (Procambarus). The common inhibitor (CI) in each thoracic segment has a medial or slightly contralateral soma near the ganglion's posterior boundary, a gently curving primary neurite, an extensive ipsilateral dendritic tree, and an axon emerging through the anterior root. The stretcher closer inhibitor (SI) has a soma slightly anterior and ipsilteral to the CI's a sharply bent proximal neurite, a smaller dendritic tree, and an axon in the posterior root. The opener inhibitor (OI) lies more laterally and often posterior to the CI; its diagnoally directed neurite enters the posterior root. The inhibitors' structures were related to major neuroanatomical landmarks within the ganglion, to soma positions of excitatory motoneurons revealed by backfilling, and to soma locations of inhibitory interneurons revealed by GABA-like immunoreactivity. In their peripheral distributions to the leg muscles and in their central structures, these crayfish limb inhibitors show striking similarities with those of the locust. Crayfish and locust thoracic ganglia also show more general neuroanatomical similarities. These observations suggest that the crayfish CI, SI, and OI are, respectively, homologous with the locust CI1, CI2, and CI3. The implications of such a homology for arthropod phylogeny are discussed. PMID- 8245219 TI - Cholinergic innervation of olfactory glomeruli in the rat: an ultrastructural immunocytochemical study. AB - The ultrastructural organization of cholinergic afferents to the rat olfactory bulb (OB) was studied with the aid of choline acetyltransferase (ChAT) immunocytochemistry in electron microscopy. Particular attention has been paid to a subset of glomeruli characterized by a remarkably high density of cholinergic afferents. Numerous cholinergic terminals making symmetric or asymmetric synaptic contacts were observed in the periglomerular area. ChAT-labelled terminals have a diameter ranging from 0.3 to 1.5 micron and contain numerous clear agranular vesicles. Axo-somatic and axo-dendritic contacts were both observed in contact with several types of target neurons. Three types of cholinoceptive, noncholinergic neurons could be identified: periglomerular cells, superficial short-axon cells, and external tufted cells. Our results provide an anatomical substrate for the hypotheses concerning the complex effects of acetylcholine in the processing of sensory information in the olfactory bulb. PMID- 8245220 TI - Roles of estradiol and progesterone in regulation of hippocampal dendritic spine density during the estrous cycle in the rat. AB - We have previously shown that the density of dendritic spines on hippocampal CA1 pyramidal cells is dependent on circulating estradiol and progesterone and fluctuates naturally during the 5 day estrous cycle in the adult rat. To date, however, no detailed characterization of the roles that these hormones play in regulation of spine density has been made. In order to determine the time courses and extent of the effects of estradiol and progesterone on dendritic spine density, we have analyzed the density of dendritic spines on the lateral branches of the apical dendritic tree of Golgi-impregnated CA1 hippocampal pyramidal cells in several experiments. In summary, our findings included the following: (1) Following ovariectomy, circulating estradiol is undetectable within 24 hours; however, spine density decreases gradually over a 6 day period. (2) Spine density does not decrease any further up to 40 days following ovariectomy. (3) Treatment with estradiol alone can reverse the ovariectomy-induced decrease in spine density. (4) Spine density begins to increase within 24 hours following estradiol benzoate injection in an ovariectomized animal, peaks at 2 and 3 days, then gradually decreases over the next 7 day period. (5) Although free estradiol is metabolized more rapidly than estradiol benzoate, there is no difference in the rate of decrease in spine density following injection of either form. (6) Progesterone has a biphasic effect on spine density in that progesterone treatment following estradiol initially increases spine density for a period of 2 to 6 hours but then results in a much sharper decrease than is observed following estradiol alone. By 18 hours following progesterone treatment, spine density is decreased nearly to 6 day ovariectomy values. (7) Treatment of intact rats with the progesterone receptor antagonist, RU 486, during the proestrus phase of the estrous cycle inhibits the proestrus to estrus drop in spine density. These findings account for both the gradual increase and rapid decrease in spine density which we have previously observed during the estrous cycle and indicate that progesterone in particular may be an important factor in the regulation of rapid morphologic changes which occur naturally in the adult brain. PMID- 8245221 TI - Origins of cerebellar mossy and climbing fibers immunoreactive for corticotropin releasing factor in the rabbit. AB - Corticotropin-releasing factor (CRF) has been implicated by both anatomical and physiological techniques as a potential cerebellar transmitter or modulator. In the present experiment, with the aid of immunohistochemistry, we have described specific cerebellar afferent pathways in the rabbit in which CRF is located. CRF immunoreactive climbing fibers were present in the molecular layer throughout the cerebellum, but especially in lobules 8-9a. All inferior olivary neurons were CRF immunoreactive. In lobules 8-9a, CRF-immunoreactive mossy fibers were organized in sagittal bands. The highest density of CRF-immunoreactive mossy fiber terminals was observed in the granule cell layer of lobules 8-9a and the flocculus. No CRF-immunoreactive perikarya were located in rabbit cerebellum. The brainstem origin of CRF-immunoreactive mossy fiber terminals was suggested by numerous CRF-immunoreactive perikarya located in the medial, lateral and descending vestibular nuclei, nucleus prepositus hypoglossi, nucleus x, paramedian reticular nucleus, gigantocellular reticular nucleus, lateral reticular nucleus, and raphe nuclei. Using double label experiments, we investigated the specific CRF afferent projection to the flocculus and posterior vermis. Horseradish peroxidase (HRP) injections into the posterior vermis double labeled CRF-immunoreactive neurons in the caudal medial and descending vestibular nuclei and nucleus prepositus hypoglossi. HRP injections into the flocculus double labeled more CRF-immunoreactive neurons in the nucleus prepositus hypoglossi than in the vestibular nuclei. HRP injections into either the posterior vermis or flocculus double labeled CRF-immunoreactive neurons in the paramedian reticular nucleus, nucleus reticularis gigantocellularis, and raphe nuclei. These data suggest that CRF may play an important role in vestibularly related functions of the cerebellum. PMID- 8245222 TI - NCAM expression by subsets of taste cells is dependent upon innervation. AB - The expression of the neural cell adhesion molecule (NCAM) and distinct carbohydrate groups by cells of the taste buds of the rat vallate papilla was investigated by immunohistochemical and biochemical techniques. We employed antibodies against 1) the extracellular (mAb 3F4) and cytoplasmic (mAb 5B8) portions of the NCAM polypeptide, 2) the highly sialylated form of NCAM (mAb 5A5), 3) carbohydrate epitopes associated with glycosylated NCAM forms in the rat (mAb 2B8) or frog (mAb 9-OE) olfactory system, and also 4) the Lewisb blood group carbohydrate epitope (mAb CO431). NCAM mRNA was demonstrated by polymerase chain reaction (PCR) in samples of the vallate papilla, suggesting the presence of NCAM in cells of the taste buds. Antibodies against NCAM (mAbs 3F4 and 5B8) recognized a subset (about 20%) of cells within the vallate taste buds; fibers of the glossopharyngeal nerve, including those innervating the gustatory epithelium, were NCAM immunoreactive. Taste bud cells did not express polysialic acid (mAb 5A5), but mAb 5A5 immunoreactivity was observed on fibers of the IXth nerve, including a few that entered the taste buds. All or nearly all of the cells within the vallate taste buds were immunoreactive to mAb 2B8, whereas mAbs 9-OE and CO431 reacted with subsets of cells. The carbohydrates recognized by mAbs 2B8 and 9-OE were also abundantly expressed in the ducts and acini of the lingual salivary glands. Bilateral crush of the IXth nerve resulted in the loss of expression of all of these molecules from the gustatory epithelium. If cells of the taste bud express NCAM during their final stage(s) of differentiation, then NCAM could play a role(s) in the growth of gustatory axons toward their target epithelial cells and in the recognition between the nerve fibers and mature taste receptor cells, or among the taste bud cells themselves. PMID- 8245223 TI - Immunolocalization of different forms of neural cell adhesion molecule (NCAM) in rat taste buds. AB - Taste buds consist of approximately 100 taste cells, including three morphological types of short receptor cells which synapse on the peripheral gustatory nerves. Some of the receptor cells produce neural cell adhesion molecule (NCAM), which may play a role in formation of specific connections in this system. Antibodies directed against different forms of NCAM were utilized in an attempt to define not only the distribution, but also the type of NCAM within taste buds. Within each taste bud approximately 10% of the taste cells exhibit abundant immunoreactivity for 180 kD (ld) or 140 kD (sd) forms of NCAM (i.e., those with an intracellular domain) along virtually the entire surface of the cell. Ultrastructural analysis reveals that these abundantly immunoreactive taste cells are of the intermediate morphological type, although not all of the intermediate taste cells within any bud are immunoreactive. In addition, the ultrastructural studies show that punctate (ld/sd) NCAM-immunoreactivity occurs on the membranes of taste cells and nerve fibers throughout each taste bud. The embryonic form of NCAM (E-NCAM), rich in polysialic acid residues, is present only in association with nerve fibers and other unidentified elongate, thin profiles of a few taste buds. The nerve plexus beneath the gustatory epithelium is also rich in NCAM-immunoreactivity. These fibers occasionally reveal immunoreactivity indicative of only the 120 kD (ssd) form of NCAM, typical of glial cells. PMID- 8245224 TI - Amphibian-specific regulation of polysialic acid and the neural cell adhesion molecule in development and regeneration of the retinotectal system of the salamander Pleurodeles waltl. AB - Antibodies specific to the neural cell adhesion molecule (NCAM-total), the 180 x 10(3) M(r) component of NCAM (NCAM-180) and polysialic acid (PSA) were used in immunohistochemistry and Western blots to detect the spatiotemporal dynamics of these molecules in development and regeneration of the retinotectal system of Pleurodeles waltl. NCAM-total and NCAM-180 are continuously expressed in the retina, optic nerve, and tectum of the developing and adult salamander. This is also found for the 140 x 10(3) M(r) component of NCAM in Western blots of the retina. In the larval retina, PSA is present in the inner plexiform layer (IPL) and a few cells in all nuclear layers. At metamorphosis, PSA expression in the retina strongly increases in the layer of cone photoreceptor somata. Several cells in the inner nuclear layer and Muller cell processes also begin to express PSA. This pattern persists into adulthood. The optic nerve and the tectum are strongly PSA-immunoreactive throughout development. In the adult optic nerve and optic fiber pathway in the brain, PSA expression is selectively downregulated. In the crush-lesioned adult optic nerve, regenerating fibers are NCAM-180-positive but PSA-negative. This demonstrates a molecular difference between growing nerve fibers of Pleurodeles in development and in regeneration. PSA regulation is closely correlated with metamorphosis, thus suggesting that PSA expression may be under hormonal control. Some aspects of PSA and NCAM isoform expression patterns in the retinotectal system of salamanders differ considerably from that of other vertebrates. The sustained expression of NCAM isoforms in adult salamanders might be due to secondary simplification (paedomorphosis). PMID- 8245225 TI - Immunocytochemical localization of neuropeptide FF (FMRF amide-like peptide) in the hypothalamo-neurohypophyseal system of Wistar and Brattleboro rats by light and electron microscopy. AB - Neuropeptide FF (F8Famide, FMRFamide-like, or morphine modulating peptide) immunoreactivity was localized by light and electron microscopy in the hypothalamo-neurohypophyseal system of Wistar and Brattleboro rats. In Wistar rats neuropeptide FF was present in part of the magnocellular neurones of the paraventricular and supraoptic nuclei in which it was coexpressed with vasopressin. Neuropeptide FF containing fibres were present in the paraventricular and the supraoptic nuclei, and in the central part of the neural lobe. At the electron microscopic level, neuropeptide FF containing nerve terminals in the neural lobe formed synaptoid contacts exclusively with pituicytes. No neuropeptide FF containing neurovascular contacts or contacts with other neuronal structures were observed. In contrast with Wistar rats, neuropeptide FF was almost completely absent in cell bodies of the paraventricular and supraoptic nuclei, and in fibres of the neural lobe in Brattleboro rats. Only a few solitary cells could be observed in these structures. The present results demonstrate that neuropeptide FF coexists with vasopressin within the hypothalamo-neurohypophyseal system. As we did not observe neuropeptide FF containing neurovascular contacts, neuropeptide FF containing nerve terminals probably have a local function within the neural lobe. Neuropeptide FF may be involved in the modulation of oxytocin and vasopressin release, with the pituicyte as an intermediate cell. PMID- 8245226 TI - Presence and biological activity of a GnRH-like factor in the nervous system of Helisoma trivolvis. AB - Gonadotropin-releasing hormone (GnRH) constitutes a family of neuropeptides found throughout the vertebrates. Although a GnRH-like peptide has also been isolated from yeast (alpha-mating factor), the presence of GnRH has not been clearly demonstrated in invertebrate phyla. In this study, we tested the hypothesis that GnRH-like peptides are present and functional in the central nervous system (CNS) of the gastropod mollusc, Helisoma trivolvis. The presence of a GnRH-like peptide was examined by three methods: (1) in immunofluorescence studies with four different antibodies generated against several GnRH peptides, select neurons and putative neurosecretory cells were specifically and consistently labelled throughout the CNS; (2) reverse-phase high performance liquid chromatography (HPLC) and radioimmunoassay (RIA) analysis revealed a GnRH-like factor which co migrates with mammalian (m)GnRH; and (3) in bioactivity experiments, extracts of Helisoma trivolvis CNS mimicked GnRH in stimulating gonadotropin release from dispersed goldfish pituitary cells in static culture. Two functional assays were carried out to examine the potential biological roles of GnRH-like peptides in Helisoma. (1) Intracellular recordings of left-parietal and visceral ganglion neurons revealed diverse electrophysiological responses to mGnRH. These effects were attenuated by a mGnRH antagonist. (2) Addition of mGnRH arrested neurite outgrowth in a subpopulation of dissociated embryonic Helisoma neurons in culture. Taken together, these results strongly suggest that a mGnRH-like peptide is an important neuropeptide in Helisoma. A hypothesis is presented that GnRH like peptides may be ancient factors that are conserved both structurally and functionally in the evolution of animals. PMID- 8245227 TI - Antennular projections to the midbrain of the spiny lobster. III. Central arborizations of motoneurons. AB - The central organization of antennular motoneurons in the brain of the spiny lobster, Panulirus argus, was analyzed by combining biocytin backfills with serial reconstructions of the antennular nerves and the brain. Eighty-nine to 99 antennular motoneurons occur in each hemibrain. The somata of the motoneurons are distributed in a consistent pattern in two complex soma clusters, the ventral paired mediolateral cluster of the deutocerebrum and the dorsal unpaired median cluster of the tritocerebrum. The motoneurons arborize ipsilaterally in the lateral and median antennular neuropils and the tegumentary neuropil. The backfills indicate a minimum of five morphological types of motoneurons with different arborization patterns. The innervation pattern of the motoneurons, together with previously reported innervation patterns of antennular sensory afferents, suggest that the lateral antennular neuropil is a lower motor center driving local antennular reflexes in response to chemical and mechanical stimulation of the antennule, whereas the median antennular neuropil is a lower motor center for equilibrium responses. PMID- 8245228 TI - Optic nerve degeneration induces the expression of MHC antigens in the rat visual system. AB - The brain has long been considered to be an immunologically privileged site. However, privilege is not absolute, as has been shown by the inability of foreign tissue grafts to survive indefinitely in the brain. The rejection of this tissue is accompanied by the upregulation of major histocompatibility complex (MHC) antigen expression. Therefore it is essential to define conditions that influence the expression of these antigens in the brain, especially since such a definition may further the understanding of disease processes that lead to the autoimmune destruction of the central nervous system. Here we show that both MHC class I and class II antigens are expressed within 1 or 2 days of eye removal by cells showing the morphological characteristics of microglia. Expression is seen along the optic pathway and within the brainstem centers to which optic axons project. In the early stages of the reaction, MHC class I antigen expression is seen throughout the optic pathway, including the terminal distribution areas of the subcortical visual centers, while MHC cells class II are localised mainly to degenerating myelinated fiber systems. These changes are not accompanied by any alteration in the integrity of the blood-brain barrier. During the second week postlesion, class I positive cells are found beyond the confines of the degenerating pathways, while class II positive cells are seen within regions such as the stratum griseum superficiale of the superior colliculus, where few myelinated axons are present. There is subsequent diminution of MHC positive cells, but a small number of cells are still seen 60 days post-lesion. Focal lesions within the eye show that at early survival times, while class I MHC positive cells are distributed throughout the nerve, class II positive cells are largely absent from the unmyelinated segment of the nerve. Retrograde changes in the retina after nerve section are accompanied only by MHC class I antigen expression. These observations show that neural degeneration is accompanied by a rigid sequence of events involving expression of MHC antigens by microglia. If foreign antigens were present in the brain while these events were taking place, it is possible that such antigens would be recognised and destroyed by the host immune system. PMID- 8245229 TI - Subacute spongiform encephalopathies--the transmissible brain amyloidoses: a comparison with the non-transmissible brain amyloidoses of Alzheimer type. PMID- 8245230 TI - Pathological characteristics of mucopolysaccharidosis VI in the rat. AB - The histological and electron microscopical characteristics of the pathology of rats with arylsulphatase B-deficient mucopolysaccharidosis (mucopolysaccharidosis VI; MPS VI) were investigated. In affected animals, intracytoplasmic vacuoles were prominent in chondrocytes, the macrophage system, cardiac valve fibroblasts, cornea, connective tissues, vascular smooth muscle cells and uterine stromal cells. Tissues containing glucosaminoglycans stored in lysosomes were positive to Mowry's colloidal iron and alcian blue stains. By electron microscopy, the lysosomes were seen to be distended by electron lucent or fine fibrillary storage material, and lysosomal storage was also detected in the endothelial cells of the arteries and cornea. In the central and peripheral nervous system abnormalities were restricted to the connective tissue. Lesions in the affected rats resembled those described in human and feline mucopolysaccharidosis VI. These results indicate that MPS VI of the rat may be a useful animal model for human MPS VI (Maroteaux-Lamy syndrome). PMID- 8245231 TI - The pathology of experimental Mycoplasma californicum mastitis in ewes. AB - Mastitis was reproduced experimentally by inoculating the lactating mammary glands of 12 ewes with Mycoplasma californicum. Large numbers of mycoplasmas were recovered from the milk. The mastitis had a lobular distribution and was characterized in the early stages by degeneration of the secretory epithelium and intense leucocytic infiltration of alveolar and ductal lumina. Later there was widespread alveolar involution, and lymphohistiocytic infiltration and fibrosis of the interstitium. A monoclonal antibody-based immunohistochemical technique detected M. californicum antigen in alveolar and ductal lumina but not in epithelium or interstitial tissue. This report is the first description of the pathological features of M. californicum mastitis in ewes. PMID- 8245232 TI - Equine pituitary adenoma: a functional and morphological study. AB - Clinico-pathological correlations in horses with pituitary adenomas are poorly understood. This paper describes the functional and morphological features of five cases of equine pituitary adenoma and of a case of multinodular pituitary hyperplasia. New findings reported include immunoreactivity for beta-lipotropin (beta-LPH), beta-melanocyte-stimulating hormone (beta-MSH), gamma 3-MSH, prolactin (PRL), and follicle-stimulating hormone (FSH) in neoplastic cells of the pituitary adenoma; and, in the multinodular hyperplasia, beta-LPH, beta endorphin (beta-END), alpha-MSH, beta-MSH, gamma 3-MSH and FSH immunoreactivity. It is suggested that the equine pituitary syndrome does not correspond to human Cushing's disease, as generally accepted, but is related to the overproduction of several pro-opiomelanocortin (POMC)-derived peptides by the cells of the tumour or hyperplastic nodules. PMID- 8245233 TI - The isolation and characterization of camel (Camelus dromedarius) immunoglobulin classes and subclasses. AB - Immunoglobulins G and M were prepared from camel sera by ammonium sulphate precipitation, gel filtration and fast protein liquid chromatography (ion exchange). Heavy and light chains were identified by polyacrylamide gel electrophoresis and Western blotting. IgG subclasses were isolated by DEAE ion exchange chromatography and shown to have different electrophoretic motilities. No molecules corresponding to IgA were detected. Rabbit anti-serum against camel IgG was generated and its specificity shown by Western blotting. PMID- 8245234 TI - Calcification of the urinary bladder in the wild cotton rat (Sigmodon hispidus). AB - Calcification of the urinary bladder epithelium was observed in 19 of 30 and 18 of 30 wild cotton rats from control and petrochemical-contaminated sites, respectively. The rats in the two sites did not differ significantly in respect of serum calcium and phosphorus concentrations. The calcification was considered to be dystrophic in nature. An unidentified factor common to both control and petrochemical-contaminated sites was considered to be responsible for this syndrome. PMID- 8245235 TI - Low nickel diet: an open, prospective trial. AB - BACKGROUND: Nickel-sensitive patients may experience persistent dermatitis even if they avoid cutaneous contact with nickel-plated items. OBJECTIVE: The purpose of the study was to determine whether reduced nickel intake in food reduces the activity of dermatitis in selected nickel-sensitive persons. METHODS: Ninety nickel-sensitive patients who had a flare of dermatitis after oral challenge with 2.5 mg of nickel but had no reaction to a placebo were instructed to adhere to a low-nickel diet. RESULTS: Fifty-eight of the 90 patients benefited in the short term from the diet, whereas 15 others had possible benefit. Seventeen patients did not benefit in the short term. Fifty-five patients who adhered to the diet for at least 4 weeks, and whose dermatitis had cleared or improved at the end of this time, responded to a questionnaire follow-up 1 to 2 years later. Forty of these patients had long-term improvement of their dermatitis. Patients with strongly positive patch tests to nickel had less benefit from the diet than patients with moderately positive patch tests. CONCLUSION: Reduction of the dietary intake of nickel may benefit some nickel-sensitive patients. PMID- 8245236 TI - A randomized, double-blind, placebo-controlled trial of ketoconazole 2% shampoo versus selenium sulfide 2.5% shampoo in the treatment of moderate to severe dandruff. AB - BACKGROUND: Ketoconazole is highly effective against the yeast Pityrosporum ovale, an organism believed to be involved in the pathogenesis of dandruff. OBJECTIVE: Our purpose was to evaluate the safety and effectiveness of ketoconazole 2% shampoo versus selenium sulfide 2.5% shampoo and placebo shampoo in patients with moderate to severe dandruff. METHODS: Features assessed included adherent and loose dandruff scores, presence or absence of irritation, itching, yeast cells, and global improvement rating by the investigator. RESULTS: A total of 246 patients were included. Mean total adherent dandruff score declined throughout the treatment period with both ketoconazole 2% and selenium sulfide 2.5% shampoos significantly better than placebo at all visits. Ketoconazole was statistically superior to selenium sulfide at day 8 only (p = 0.0026). Both medicated shampoos were significantly better than placebo for reducing irritation and itching. Of the nine adverse experiences reported during the treatment phase, all involved patients treated with selenium sulfide 2.5% shampoo. CONCLUSION: Both ketoconazole 2% shampoo and selenium sulfide 2.5% shampoo are effective in the treatment of moderate to severe dandruff; however, ketoconazole 2% shampoo appears to be better tolerated. PMID- 8245237 TI - PUVA, UVB, psoriasis, and nonmelanoma skin cancer. AB - This article reviews the English-language literature on the risk of nonmelanoma skin cancer from phototherapy (UVB) or photochemotherapy (PUVA) in the treatment of psoriasis. The evolution of the debate on the carcinogenic potential of these treatment is discussed, particularly with reference to PUVA. Available data indicate that therapeutic UVB has a low risk of producing cutaneous cancers, with the possible exception of those on male genital skin. However, there is a definite cutaneous carcinogenic risk from oral PUVA. The major associated risk factors are reviewed. We provide recommendations for the assessment and treatment of patients to minimize this risk. PMID- 8245238 TI - Exacerbation of porokeratosis: a sign of immunodepression. PMID- 8245239 TI - Human papillomavirus type 1 DNA in verrucous carcinoma of the leg. PMID- 8245240 TI - Prevention of blisters in dystrophic recessive epidermolysis bullosa with cyclosporine. PMID- 8245241 TI - Linear hyperpigmentation overlying veins in association with collagen vascular disease. PMID- 8245242 TI - Successful treatment of disseminated verrucae vulgares with interferon-beta. PMID- 8245243 TI - Digital angioleiomyoma: an uncommon neoplasm. PMID- 8245244 TI - Prevention of skin cancer in xeroderma pigmentosum: the physician as advocate. PMID- 8245245 TI - Cutaneous T-cell lymphoma mimicking porokeratosis of Mibelli. PMID- 8245246 TI - Effect of cyclosporine on genital psoriasis and lichen planus. PMID- 8245247 TI - Primary neuritic leprosy. PMID- 8245248 TI - Delayed cutaneous ulcerations arising at sites of prior parenteral drug abuse. PMID- 8245249 TI - Bilateral renal cell carcinoma in the Birt-Hogg-Dube syndrome. PMID- 8245250 TI - On rifampin treatment of psoriasis. PMID- 8245251 TI - Is the screening patch test tray still worth using? PMID- 8245252 TI - Biblical leprosy. PMID- 8245253 TI - Basal cell carcinoma arising in a fibroepithelial polyp. PMID- 8245254 TI - Unilateral dermatomal pigmentary dermatosis: a variant dyschromatosis? PMID- 8245255 TI - The rationale of the ABCDs of early melanoma. PMID- 8245256 TI - Angiotensin-converting enzyme inhibitors. PMID- 8245257 TI - Angiotensin-converting enzyme inhibitors. PMID- 8245258 TI - Facial cutaneous reconstructive surgery: facial flaps. AB - Basic principles of aesthetics and function important for successful facial cutaneous reconstructive surgery with local flaps include the use of appropriate tissue donor sources, knowledge of the mechanisms, dynamics, and effects of tissue movement, and techniques for scar camouflage. Useful flaps for defects in each of the facial cosmetic units are reviewed with these principles used as guidelines. PMID- 8245259 TI - Lymphoma versus pseudolymphoma of the skin: gene rearrangement study of 21 cases with clinicopathologic correlation. AB - BACKGROUND: Diagnosis of cutaneous lymphoma in the absence of systemic lymphoma may be difficult. Reactive lymphoid lesions can mimic lymphoma clinically and histologically and have been designated pseudolymphomas. OBJECTIVE: Our purpose was to analyze lymphoid gene rearrangements in cutaneous lymphoproliferative lesions and to correlate these findings with the histologic, immunophenotypic, and clinical profile. METHODS: We examined 21 cases of lymphoproliferative lesions that developed in skin and performed molecular rearrangement analysis of T-cell receptor and immunoglobulin genes. We examined identical tissues by histologic and immunophenotypic criteria and conducted follow-up clinical evaluation of all patients. RESULTS: Clonal rearrangements of immunoglobulin (seven cases) or T-cell receptor (two cases) gene were detected in 9 of 21 patients. No specific histologic or immunophenotypic feature was consistently associated with a clonal lymphoid gene rearrangement. Systemic lymphoma developed in one patient in whom a clonal rearrangement within the immunoglobulin gene was identified. CONCLUSION: Gene rearrangement analysis may be helpful in differentiating primary cutaneous lymphoma from pseudolymphoma. The chronic clinical course of patients with clonal lymphoid gene rearrangements supports a lack of correlation between clonality and biologic aggressiveness. PMID- 8245260 TI - Diagnosis and incidence of delayed pressure urticaria in patients with chronic urticaria. AB - BACKGROUND: The incidence of delayed pressure urticaria (DPU) may have been underestimated, particularly in patients with widespread wheals of concurrent idiopathic urticaria, because of difficulty in recognizing the association between pressure-related swellings and the preceding physical stimulus. Diagnostic tests for DPU have not been done routinely in studies of urticaria and have not been compared. OBJECTIVE: Our purpose was to establish the incidence of DPU and other physical urticarias in patients with chronic urticaria and to compare two reproducible pressure challenge tests. METHODS: One hundred thirty five patients were tested for immediate dermographism, for DPU and, when indicated by the history, for cholinergic and/or cold urticaria. In patients with pressure-related symptoms, pressure challenge testing with a dermographometer (100 gm/mm2) was compared with a test that involves the application of 1.5 cm diameter weighted rods and more closely resembles a naturally occurring pressure stimulus. RESULTS: Physical urticarias were present in 96 patients (71%). Thirty patients (22%) had immediate dermographism, 50 (37%) had DPU, 15 (11%) had cholinergic urticaria, and 3 (2%) had cold urticaria. A 70-second pressure challenge with the dermographometer gave results comparable to the use of the weighted rods. CONCLUSION: DPU appears to be more common in chronic urticaria than previously reported and is present in some patients who do not report pressure-related wheals. PMID- 8245261 TI - A survey of attitudes, beliefs, and behavior regarding tanning bed use, sunbathing, and sunscreen use. AB - BACKGROUND: Although cosmetic tanning and unprotected solar exposure are common, little is known about general attitudes, beliefs, and behavior regarding sunbathing, sunscreen use, and tanning salon use. OBJECTIVE: We sought to determine the frequency of UV exposure in a select sample and to assess the knowledge and beliefs of the effects of UV irradiation. METHODS: A written, anonymous questionnaire was distributed to a sample of 477 persons in a shopping mall, at a social gathering, and on a vacation cruise ship. The instrument explored demographic information, sunscreen use, sunbathing habits, tanning bed use, and cutaneous solar effects. RESULTS: Forty-two percent of respondents seldom or never used sunscreen, and 33% sunbathed at least once a week. Although the three sample populations differed in education, sunbathing habits, sunscreen use, and tanning bed use, they were equally informed about UV light hazards. Compared with those who had not used tanning beds, tanning bed users were more likely to be female and more knowledgeable about the long-term effects of UV. Tanning beds were most commonly used in tanning or hair salons, (mean 23 +/- 7 minutes at 2.3 +/- 1.1 times per week). Reported positive psychologic sequelae from tanning bed use were more common than negative physical sequelae. At least 10% would continue to use tanning beds if these were proved to cause skin cancer. CONCLUSION: In this select sample, sunbathing and tanning bed use were common. No group surveyed universally practiced sun protection and avoidance. Clientele of tanning beds may be aware of the damaging effects of the sun, but may not be aware that tanning bed use is associated with skin damage. PMID- 8245262 TI - Topical administration of hexadecylphosphocholine in patients with cutaneous lymphomas: results of a phase I/II study. AB - BACKGROUND: Hexadecylphosphocholine is a new antineoplastic drug that inhibits tumor cell growth directly and, in addition, might have immunoregulatory properties. OBJECTIVES: We investigated the topical application of this phospholipid in patients with cutaneous lymphoma. METHODS: Twenty-four patients with histologically documented cutaneous lymphoma were treated for 8 weeks. Lesions that responded to treatment were biopsied and evaluated histologically. RESULTS: Of 15 patients with cutaneous T-cell lymphomas, 12 were evaluable. Two complete remissions, four partial remissions, and one minor remission were observed. Of seven patients with B-cell lymphomas, six were evaluable. One complete remission, three partial remissions, one case of stable disease, and one case of progressive disease were seen. However, histologic monitoring demonstrated only a partial clearing of infiltrating lymphocytes in lesions that showed a partial or complete response clinically. Both patients with lymphomatoid papulosis had complete clearing of the lesions clinically. An objective response rate (partial and complete response) of 56% (10/18) was achieved in the patients with cutaneous lymphoma who were treated in this study. CONCLUSION: Hexadecylphosphocholine appears to be effective topically in the treatment of some cases of cutaneous lymphomas. PMID- 8245263 TI - Herpes zoster: daily marking of new vesicles in therapeutic studies. A clinical method for objective assessment of the end of the eruptive phase. AB - BACKGROUND: The efficacy of a therapeutic agent must be evaluated by objective criteria. However, in herpes zoster (HZ) studies there has been no generally accepted objective clinical criterion. OBJECTIVE: Our purpose was to establish a clinical method for determining objectively the point in time at which the eruptive phase of HZ is completed (no new vesicle formation). This point is said to be a clinical criterion for the end of viral replication in the skin and thus for measuring the efficacy of a virustatic agent. METHODS: Newly formed vesicles were marked with differently colored permanent marker pens each day. This method was evaluated by comparing the results of acyclovir therapy in two groups of patients with HZ. (Group A, no underlying malignancy; n = 9. Group B, underlying malignancy; 64% of these patients were undergoing cytostatic polychemotherapy or had immunodeficiency; n = 22). RESULTS: In both groups, acyclovir stopped the eruption of new vesicles within 1.8 and 2.8 days, respectively (not statistically significant). Group B showed a tendency toward more protracted hematogenous dissemination and a longer duration of therapy. The total duration of the eruptive phase depended solely on the length of the interval between the onset of the HZ and the beginning of therapy. CONCLUSION: The method of marking new vesicles is independent of laboratory facilities, simple, and cost effective; in addition, this method is suitable for statistical evaluation. It is thus superior to other clinical methods for objective assessment of the progression of HZ. PMID- 8245264 TI - Pretibial epidermolysis bullosa: a clinicopathologic study. AB - BACKGROUND: Pretibial epidermolysis bullosa (PEB) is a rare variant of dystrophic epidermolysis bullosa (DEB) that predominantly involves the pretibial skin. OBJECTIVE: We report 19 cases of DEB with pretibial predilection and our findings on anchoring fibrils. METHODS: Patients with blisters and scars that primarily involved the pretibial were selected. Blisters and noninvolved skin were studied by light and electron microscopy. Anchoring fibrils were quantified by morphometry. RESULTS: There were 19 patients from 13 families. The inheritance was autosomal dominant in 10 families and sporadic in one family, and sibling involvement was present in two families. Nail dystrophy was noted in all patients. Pruritus was a common feature. Extensive prurigo occurred in four patients. Eight patients had skin lesions that exclusively affected the legs and were diagnosed as PEB. The remaining patients also showed some albopapuloid or hypertrophic scars at sites other than the leg, features seen in the albopapuloid or Cockayne-Touraine types. These were labeled as albopapuloid PEB or Cockayne Touraine PEB. Common ancestry and various combinations of PEB, with albopapuloid or Cockayne-Touraine type of EB, were noted in some families. Compared with normal controls, the anchoring fibrils were rudimentary and sparser in both lesional and nonpredilected normal skin, although in the latter the changes were less marked in PEB than in the albopapuloid or Cockayne-Touraine types. CONCLUSION: Common ancestry may contribute to the higher incidence of DEB, especially PEB, in Tainan, Taiwan. The abnormalities of anchoring fibrils were not restricted to the predilected site in PEB and did not distinguish PEB from other types of DEB. PMID- 8245265 TI - Randomized trial comparing cryopreserved cultured epidermal allografts with hydrocolloid dressings in healing chronic venous ulcers. AB - BACKGROUND: Cultured epidermal allografts have been successfully used to treat a variety of wounds. Their postulated mechanism of action is through release of cytokines that stimulate epithelialization. On the basis of previous experience we expected ulcers treated with cryopreserved cultured allografts (CCAs) to be healed by 6 weeks. Hydrocolloid dressings (HCDs) have also been reported to be effective in the treatment of venous ulcers. OBJECTIVE: Our purpose was to compare the effectiveness of CCAs with HCDs in healing chronic venous ulcers. METHODS: Forty-three patients with 47 ulcers were enrolled in a randomized controlled trial. Ulcers not healed by 6 weeks were changed to the other treatment. RESULTS: No difference in the number of healed ulcers between the two groups was observed at 6 weeks. Healing rate, percent reduction of initial ulcer size, and radial progression toward wound closure were significantly greater for CCAs than for HCDs. Pain relief was not significantly different. CONCLUSION: CCAs achieve more rapid healing and greater reduction in ulcer size than HCDs. PMID- 8245266 TI - How common is the atypical mole syndrome phenotype in apparently sporadic melanoma? AB - BACKGROUND: Although patients from some families with the atypical mole syndrome (AMS) are predisposed to melanoma, it is not known how frequently this underlies the apparently sporadic presentation of melanoma. OBJECTIVE: This study was designed to estimate the frequency of the AMS (dysplastic nevus or FAMMM syndrome) phenotype in a population-based study of patients with melanoma in the United Kingdom and to determine the prevalence of the phenotype in the relatives of the patients with AMS. METHODS: The nevi of patients with melanoma and controls in a case-control study, and the nevi of some relatives of patients with AMS, were examined. An AMS scoring system was used to define the AMS phenotype. The familiarity of the AMS phenotype was then determined by screening first degree relatives of persons with the AMS phenotype. RESULTS: Forty of 266 (15%) of patients with melanoma had the AMS phenotype compared with 7 of 305 (2%) of the controls (odds ratio 7.5, 95% confidence interval 3.4-16.8). Screening of relatives of patients with melanoma who had the AMS phenotype identified the same phenotype within the families, providing evidence that the AMS phenotype in patients with melanoma is predictive of the same phenotype in relatives, consistent with so-called type D1 AMS. CONCLUSION: The AMS phenotype is a potent risk factor for cutaneous melanoma and is present in 15% of patients. Melanoma in the United Kingdom is more common in women than in men, but the AMS phenotype was more frequent in men in this study. It is our hypothesis that the effects of the putative AMS gene are diluted by environmental factors in U.K. women. Screening of relatives of patients with melanoma who have the AMS phenotype may identify persons at increased risk of melanoma. PMID- 8245267 TI - Nevus of Ota: treatment with high energy fluences of the Q-switched ruby laser. AB - BACKGROUND: The nevus of Ota is a benign dermal melanocytic lesion that has previously proved difficult to treat. Recently, the Q-switched ruby laser has been reported to be successful in treating benign pigmented lesions and tattoos. OBJECTIVE: Our study evaluates the treatment of 16 patients with nevus of Ota with the Q-switched ruby laser (694 nm). METHODS: Sixteen patients with nevus of Ota were treated with the Q-switched ruby laser with a pulse width of 28 nsec and energy fluences ranging from 7.5 to 10 J/cm2. Response to treatment was assessed by an independent investigator with photographs. RESULTS: The average number of treatments was 3.8 per patient. After two treatments, 44% of patients showed a 50% or greater improvement. After three treatments, 85% of patients showed a 50% or greater improvement; after four treatments, 100% of patients showed 50% or greater improvement. No patients had permanent textural changes or scarring. CONCLUSION: High-energy fluences of the Q-switched ruby laser lead to significant improvement without scarring of nevus of Ota after a few treatments. PMID- 8245268 TI - Introduction to special section: clinical research in smoking cessation. AB - Although tobacco smoking is becoming less common in the United States, it remains perhaps the most important behavioral problem treated by psychologists. Research interest in smoking has greatly increased among psychologists, but success rates of formal treatment programs for smoking cessation have remained stubbornly low. This article introduces a series of 9 reviews that examine the current status of several diverse areas of research in smoking cessation and provide directions for future efforts in these areas in an effort to promote more innovative investigations. PMID- 8245269 TI - Smoking cessation treatment: any progress? AB - This article argues that, although smoking rates are declining and quit rates are rising, research on behavioral approaches to smoking cessation is stagnant. Despite an increase in publications on smoking cessation, few innovative approaches have been introduced in the past decade. Furthermore, average treatment outcomes have not improved over this period. Treatment innovations may come from research that aims at fundamental new insights regarding smoking behavior and smoking behavior change rather than from incremental improvements of current approaches. PMID- 8245270 TI - Why some people smoke and others do not: new perspectives. AB - Because initial reinforcement consequences set the stage for subsequent nicotine use, elucidation of the contribution of environmental and inherited factors is crucial to an understanding of nicotine dependence as well as of individual differences in susceptibility to cigarette smoking. A review of some recent animal research and laboratory studies of smokers and never-smokers suggests that vulnerability to nicotine dependence is related to high initial sensitivity to nicotine and that the development of acute pharmacodynamic tolerance in these individuals--particularly to effects of nicotine that are toxic or aversive--may be an adaptation for protecting homeostasis. PMID- 8245271 TI - Assessing smoking patterns and motives. AB - This article focuses on measures used to assess smoking patterns and motives. Self-report typology scales demonstrate a remarkably stable factor structure and good reliability. However, self-monitoring studies suggest that typology scales do not accurately assess smoking patterns and do not provide a good basis for treatment planning. Nevertheless, typology scores are correlated with variables such as smoking rate, craving, and cessation outcomes. These correlations may largely be accounted for by a common underlying factor, here labeled smoking drive, which may serve as a measure of dependence. Finally, the article briefly introduces new methods for assessing smoking patterns using palm-top computers to track smoking and other stimuli in real time. These methods may provide more valid assessment of smoking patterns. PMID- 8245272 TI - Nicotine delivery kinetics and abuse liability. AB - It is well established that nicotine meets all criteria of a highly addictive drug. However, as recognized by the U.S. surgeon general, the nicotine delivery system itself is an important determinant of the toxic and addictive effects engendered by nicotine use. Therefore, altering the form of nicotine dosing may allow for selective therapeutic action in efforts to develop safer and less addictive nicotine replacement therapies. While it is the case that initial tobacco use often escalates to compulsive use accompanied by tolerance and physical dependence, this is not usually observed with nicotine replacement therapies. These observations are consistent with laboratory data indicating that (a) nicotine polacrilex and transdermal systems deliver nicotine more slowly and at lower dose levels than tobacco-based forms, and (b) human data suggesting that the abuse liability of these systems is substantially lower than that of the tobacco-based nicotine delivery systems. Because the drug dosage form can be systematically manipulated and evaluated, further research in developing alternative nicotine delivery forms may hold substantial promise in the treatment of tobacco dependence. Psychological research methods can play an important part in their evaluation. PMID- 8245273 TI - Pharmacotherapy for smoking cessation: unvalidated assumptions, anomalies, and suggestions for future research. AB - This article questions several assumptions about the rationale for pharmacological therapies for smoking cessation, including whether (a) future smokers will be those more dependent on nicotine and thus in greater need of nicotine replacement or other pharmacotherapy, (b) transdermal nicotine and nicotine gum work by reducing withdrawal symptoms, and (c) clonidine works by decreasing sympathetic arousal. After describing currently available pharmacotherapies, the article also describes several unexpected findings that need to be taken into consideration by clinicians: (a) transdermal nicotine is effective when given without psychological therapy, (b) transdermal nicotine and nicotine gum do not consistently decrease postcessation weight gain, (c) high level of nicotine dependence does not consistently predict better response to transdermal nicotine, and (d) clonidine is effective in women but not in men. The article poses other questions for future research. PMID- 8245274 TI - Weight gain following smoking cessation. AB - Smoking generally suppresses body weight below "normal," and smoking cessation allows weight to return to normal. This weight gain following cessation appears to be due to a transient increase in eating coupled with the removal of acute metabolic effects of each cigarette, with no change in physical activity. Nevertheless, tobacco smoke (and specifically nicotine) does not appear to be simply either an anorectic or a thermogenic agent. Although there may be no easy explanation for the effects of smoking on energy balance, the most parsimonious explanation may be that smoking lowers body weight "set point" and cessation raises set point. The transient changes in eating are therefore secondary to the changes in body weight set point. This notion is supported by animal research with nicotine as well as with other drugs, and it is also supported less directly by the pattern of changes observed with changes in smoking status among humans. A set-point explanation for weight gain after smoking cessation may also help explain the lack of success of interventions designed to prevent this weight gain. PMID- 8245275 TI - Helping hospitalized smokers quit: new directions for treatment and research. AB - To date, relatively little work has been done to develop or evaluate effective inpatient quit-smoking treatment programs. However, there is growing interest in programs that motivate and assist the hospitalized smoker to quit smoking and remain abstinent. This article presents the rationale for hospital-based smoking treatment programs and introduces a practical minimal-contact treatment model based on extensive studies of primary-care-based and self-help interventions, on a limited number of inpatient studies, and on several prominent theories of health behavior change. Recent controlled and exploratory studies of inpatient interventions are reviewed, including programs for the general medical population and programs for special patient groups such as patients hospitalized for cardiovascular disease, pulmonary disease, cancer, and drug or alcohol dependency. Major findings are summarized, and recommendations are given for future treatment and research. PMID- 8245276 TI - Self-help interventions for smoking cessation. AB - Self-help interventions for smoking cessation are an important bridge between the clinical and public health approaches to smoking cessation. The current literature on self-help interventions is encouraging but incomplete. Although their quit rates are lower than those of more intensive programs, self-help interventions could have a large public health impact because of their potential for widespread distribution. Studies comparing self-help to more intensive treatment suggest that long-term cessation rates for self-help programs are potentially as high as rates for face-to-face interventions, with lower quit rates for self-help programs that are likely due to differences in program adherence. Tailored materials and personalized adjuncts (e.g., written feedback or telephone counseling) that promote program adherence may increase cessation rates. PMID- 8245277 TI - Using research to guide culturally appropriate drug abuse prevention. AB - Several ways in which research can be an important tool in creating culturally appropriate prevention programs are suggested. To illustrate cultural variability, we present data from a study of 5th-grade African-American and European-American students. These data describe rates of early initiation, levels of exposure to identified risk factors, and degree to which identified risk factors predict substances initiated. Results indicate that European-American Ss have higher rates of tobacco and alcohol initiation, that the same risk factors predict the variety of substances initiated for African-American and European American Ss, and that several differences exist in the level of exposure to risk factors between the 2 groups. Implications of the findings for developing culturally appropriate drug abuse prevention are discussed. PMID- 8245278 TI - Experience-dependent neuropsychological recovery and the treatment of alcoholism. AB - Accumulating research has shown that some cognitive deficits in recently abstinent alcoholics (e.g., cognitive flexibility, acquisition of novel skills) improve only with remediation in contrast to the spontaneous, time-dependent rebound seen for other tasks. In principle, this facilitated or experience dependent recovery should enhance acquisition of the content of alcoholism treatment programs, but this relationship has yet to be tested empirically; previous research assessed recovery using only neuropsychological tasks presented by an experimenter. The current investigation focused on treatment-relevant remediation (acquisition of the content of a relapse-prevention [RP] program) using tasks administered by self-guided workbooks. Four groups of male alcoholics received pre- and posttesting. Between the two testing sessions, the groups received neuropsychological remediation tasks (n = 15), ecologically relevant tasks (n = 15), attention-placebo tasks (n = 16), or no intervention (n = 15). Results showed that exposure to both types of remediation produced significant cognitive recovery, with skills transferring to posttest neuropsychological measures and RP acquisition. Hence, cognitive remediation may facilitate alcoholism treatment. PMID- 8245279 TI - Meta-analytical review of the efficacy of nicotine chewing gum in smoking treatment programs. AB - The research on the effectiveness of nicotine gum as a treatment for smoking was reviewed through a meta-analysis of 33 studies. The differential effectiveness of experimental (nicotine gum) versus control (placebo and no gum) groups at both short- and long-term follow-up was indexed as d, the mean effect size. These effect sizes were contrasted within each brief and intensive treatments. Nicotine gum was superior to placebo and no-gum controls at both intervals with the intensive strategies, but the gum was effective only at short term for the brief treatments. Results show that the positive effects of nicotine gum in smoking treatment are a function of an interaction between the gum's pharmacological properties and the effectiveness of intensive treatment strategies. The theoretical and clinical implications of the results are discussed. PMID- 8245280 TI - Stress and unaided smoking cessation: a prospective investigation. AB - A prospective design was used to determine the outcomes associated with unaided smoking cessation and the influence of stress on cessation. Heavy smokers (N = 308) completed stress-related measures and were then recontacted at 1, 6, and 12 months. At each follow-up, they indicated their smoking status (which was confirmed by collateral report and biochemical tests) and completed several stress-related questionnaires. Results indicated that 33% of subjects smoked continuously throughout the year, 39% quit briefly but subsequently relapsed, and 15% quit (confirmed biochemically). An additional 7% reported that they had quit, but this could not be confirmed, and 6% were lost to follow-up. Compared with nonquiters, quitters reported less perceived stress, greater self-efficacy, greater use of problem solving and cognitive restructuring, and less reliance on wishful thinking, self-criticism, and social withdrawal. A model to forecast quitting was built and cross-validated. PMID- 8245281 TI - Three-year course of behavioral/emotional problems in a national sample of 4- to 16-year-olds: 3. Predictors of signs of disturbance. AB - We tested parent-reported family variables, problems, competencies, and stress as predictors of (a) academic problems, (b) school behavior problems, (c) receipt of mental health services, (d) child's need for additional help, (e) suicidal behavior, (f) police contacts, and (g) the sum of these 6 outcomes. Included in the study were 995 cases manifesting at least 1 sign of disturbance (from Outcomes a-f) and 995 matched controls from a national sample of 2,479 children assessed twice over a 3-year interval. Path analyses identified predictors that were significant across age and sex plus those specific to particular groups. The predictive models accounted for medium to large effects in Time 2 signs of disturbance. Time 2 Child Behavior Checklist scores were significantly associated with all Time 2 disturbance scores. The multiple significant risk factors associated with signs of disturbance indicated variations in pathways leading to particular signs of disturbance. PMID- 8245282 TI - Do depressed patients in different treatment settings have different levels of well-being and functioning? AB - Differences in the functioning and well-being of adult patients with current or past depressive disorder who visited clinicians of different specialties in health maintenance organizations, solo practices, or large multispecialty group practices were examined. For patients in different systems, there were no significant differences in functioning and well-being across 12 domains tested. Patients of mental health specialists had worse mental health and more limitations in social activities, whereas patients of medical clinicians had worse physical functioning, more pain, more physical/psychophysiologic symptoms, and worse health perceptions. Thus, each system of care had depressed patients with a similar functioning and well-being "burden" but specialty sectors had patients with slightly different functioning and well-being profiles, probably reflecting patient selection of type of provider. PMID- 8245283 TI - Temporal course of change of depression. AB - Two hundred fifty moderately to severely depressed outpatients were randomly assigned to 16 weeks of cognitive-behavioral therapy, interpersonal psychotherapy, imipramine plus clinical management (IMI-CM), or pill placebo plus clinical management. Two hundred thirty-nine patients actually began treatment. The most rapid change in depressive symptoms occurred in the IMI-CM condition, which achieved significantly better results than the other treatments at 8 and 12 weeks on 1 or more variables. Change over the course of treatment on variables hypothesized to be most specifically affected by the respective treatments was found only in the case of pharmacotherapy, in which imipramine produced significantly greater changes on the endogenous measure at 8 and 12 weeks. PMID- 8245284 TI - Coping modeling problem solving versus mastery modeling: effects on adherence, in session process, and skill acquisition in a residential parent-training program. AB - This trial compared two approaches used to introduce parenting skills in a residential staff training program. Fifty staff were randomly assigned to: mastery modelling in which videotaped models demonstrated new skills, coping modelling problem solving (CMPS) in which participants formulated their own solutions to the errors depicted by videotaped models, or a waiting-list control group. In both, leaders used modelling, role playing, and homework projects to promote mastery and transfer of new skills. The skills of all groups improved, but CMPS participants attended more sessions, were late to fewer sessions, completed more homework, engaged in more cooperative in-session interaction, rated the program more positively, and reported higher job accomplishment scores. These data suggest that CMPS allowing participants to formulate their own solutions may enhance adherence and reduce the resistance observed in more didactic programs. PMID- 8245285 TI - Cognitive, behavioral, and family factors in the differentiation of depressive and anxiety disorders during childhood. AB - Contribution of cognitive, behavioral, and family environment variables to the differentiation of depressive and anxiety disorders in children was explored. Fifty-nine children from Grades 4-7 (14 diagnosed with a depressive disorder, 16 diagnosed with depressive and anxiety disorders, 11 diagnosed with an anxiety disorder, and 18 nondisturbed controls) completed measures of the depressive cognitive triad, depressive cognitions, social skills, family environment, and maladaptive family messages. Results of a stepwise discriminant function analysis indicated that 2 discriminant functions composed of 7 variables from the cognitive, behavioral, and family environment domains accounted for 91% of the between-groups variance. Results suggest that depressive disorders can be distinguished from anxiety disorders on the basis of ratings of cognition, social skills, and family environment. Implications for existing research and a model of depression during childhood are discussed. PMID- 8245286 TI - Earvin "Magic" Johnson's HIV serostatus disclosure: effects on men's perceptions of AIDS. AB - The effects of celebrity self-disclosure of human immunodeficiency virus (HIV) seropositivity on perceptions of HIV and Acquired Immunodeficiency Syndrome (AIDS) were investigated. AIDS-related interest and knowledge measures were collected from 468 men before and after basketball star Earvin "Magic" Johnson's self-disclosure of HIV seropositivity. Increased interest in AIDS paralleled media coverage of the announcement, with the most substantial effects occurring within 2 weeks. Perceived impact of the disclosure was greatest among African American men and men who had not previously known someone with HIV-AIDS. Celebrity self-disclosure appears to affect perceptions through mechanisms similar to those involved in personally knowing someone infected with HIV. PMID- 8245287 TI - Comparative study of emotional experiencing in psychodynamic-interpersonal and cognitive-behavioral therapies. AB - Despite increased interest in the role of emotion in the process of psychotherapy, we currently lack a valid gauge of its importance in the change process. Significant sessions obtained from 13 experienced psychodynamic interpersonal and 17 experienced cognitive-behavior therapists were examined to determine the extent of affective exploration and therapists' views of these client states. Results indicate that affective experiencing is present in equivalent amounts in the change sessions of these two orientations. However, therapists' clinical views were dissimilar. Psychodynamic-interpersonal therapists viewed portions containing higher affective experiencing as more critical to the change process, whereas cognitive-behavior therapists viewed lower levels of experiencing as being therapeutically more significant. These findings suggest that, with regard to emotional experiencing, psychodynamic interpersonal therapists may share common ground with experiential therapists, whereas cognitive-behavior therapists appear to have a unique perspective. PMID- 8245288 TI - Toward a competency-based curriculum. PMID- 8245289 TI - Trends in the prescription of radiographs for comprehensive care patients in U.S. and Canadian dental schools. AB - Results of this 1992 survey are compared to results from 1977 and 1987 surveys to establish long-term trends in radiographic prescribing practices for dentulous adult comprehensive care patients in U.S. and Canadian dental schools. The major trends include a decline in prescribing both an intraoral full-mouth series and panoramic radiograph (46 percent to 19 percent to 9 percent of schools from 1977 to 1987 to 1992, respectively). These changes were complemented by an increase in prescribing a full-mouth series only (32 percent to 48 percent to 57 percent, respectively), and an increase in the use of selection criteria (2 percent to 19 percent to 26 percent, respectively). Between 1987 and 1992 there was little change in the preferred examination for the edentulous adult patient, and small changes in the preferred examination for the child patient. PMID- 8245290 TI - Practice characteristics of graduates of postdoctoral general dentistry programs. AB - A mail questionnaire was sent to 900 dentists who completed a Postdoctoral General Dentistry (PGD) program in 1985 and 900 dentists who graduated from dental school in 1985 but did not participate in a PGD program. The response rate was 57 percent. Three-quarters of the PGD graduates trained in civilian programs with the remainder being equally divided between military and Veterans Administration (VA) programs. Graduates of military programs continued to be employed by the military five years after graduation in contrast to the VA where less than 2 percent of graduates were employed after five years. The percent of dentists who are specialists or are enrolled in specialty programs five years after graduation was approximately 13 percent and the rates were similar for PGD and non-PGD dentists. The patterns of care in private practice tended to be similar for both groups; however, in institutions GPR graduates provided more treatment for special care patients and patients who were economically disadvantaged. GPR graduates also were more involved in teaching and in hospital activities. PMID- 8245291 TI - Effect of planning the desired outcome on the quality of tooth preparation. PMID- 8245292 TI - Utilization of radiographs for a state dental hygiene board examination. PMID- 8245293 TI - Management of patients susceptible to bacterial endocarditis and related infections. PMID- 8245294 TI - Case-based learning for orofacial pain and temporomandibular disorders. PMID- 8245295 TI - The oral health research work-force. PMID- 8245296 TI - Curriculum guidelines on community dental health for dental hygienists. American Association of Dental Schools. PMID- 8245297 TI - Curriculum guidelines for postdoctoral operative dentistry. American Association of Dental Schools Section on Operative Dentistry. PMID- 8245298 TI - Flash lamp-pumped dye laser treatment of telangiectasia. AB - BACKGROUND: Although many treatment modalities are effective in the treatment of essential telangiectasia, most have a risk of scarring and or incomplete obliteration of the dilated vessels. The Candela flash lamp-pumped dye laser has been regarded as highly selective for blood vessel ablation with virtually no risk of scarring. OBJECTIVE: To present a study of 182 patients who underwent this treatment. RESULTS: Good to excellent response was seen in 97.5% of these patients. No scars were seen in any of the patients studied. Patients described the degree of discomfort as mild to moderate and found the post-treatment purpura an acceptable phase of healing, if advised preoperatively. CONCLUSION: We found the laser to be safe and effective in the treatment of facial telangiectasia. PMID- 8245299 TI - Scalp lifting. An 8-year experience with over 1,230 cases. AB - BACKGROUND: This article reviews the author's 8-year experience with extensive scalp-lifting, which includes 1,230 cases. OBJECTIVE: To overview extensive scalp lifting paying particular attention to the areas of complications and the ways to reduce them. METHOD: Retrospective analysis of 1,230 cases was done. Necrosis, infection, hematoma, numbness, and scarring were evaluated. RESULTS: 1) Vertical incision occipital artery ligatory should be performed 4 to 8 weeks prior to a scalp lift. 2) Use a delay procedure prior to a frontoparietal advancement flap. 3) Use only lateral lifts on patients with prior punch-grafting. 4) Locate the temporal arteries preoperatively with Doppler ultrasonography. 5) Do not perform frontoparietal advancement flaps on patients with prior punch-grafts. CONCLUSION: Scalp lifting is an extremely effective and expedient method to treat alopecia; however, certain precautionary measures must be taken to obtain excellent, consistent results. PMID- 8245300 TI - Histiocytic reaction associated with topical aluminum chloride (Drysol reaction) AB - BACKGROUND: In the past few years, dermatologists have begun to use aluminum chloride (Drysol) as a hemostatic agent for minor surgical procedures. An unusual histiocytic reaction was noted in biopsies of skin previously treated with aluminum chloride. This reaction consisted of a proliferation of histiocytic cells that contained prominent basophilic cytoplasmic granules. OBJECTIVE: To determine the cause of this reaction and the nature of the basophilic granular material within the histiocytic cells. METHODS: Four cases are presented in which re-excised tissue previously treated with aluminum chloride were examined with special histochemistry staining and roentgen diffraction studies. RESULTS: The granules of these histiocytes stained positively with the aluminon stain, a stain specific for aluminum, although no aluminum was found using the less sensitive roentgen diffraction studies. CONCLUSION: These studies support the concept that aluminum chloride can cause a proliferative histiocytic reaction when used as a topical cauterizing agent. PMID- 8245301 TI - Dermal augmentation with fat: a reality? PMID- 8245302 TI - Autologous collagen? Lipocytic dermal augmentation. A histopathologic study. AB - BACKGROUND: Soft tissue augmentation with processed fat has been used clinically for several years. This material has been termed "autologous collagen" since it was presumed it was composed primarily of collagen and fibrous tissue from lipocyte cell walls. OBJECTIVE: To examine the histologic behavior of dermis implanted with processed fat. METHOD: Volunteers were injected with processed fat intradermally in the post auricular area and these sites were biopsied at one week, one month, and three months. RESULTS: Early biopsies revealed no intact adiopocytes but a significant inflammatory infiltrate. Subsequently biopsies revealed replacement of the inflammatory reaction with cellular fibrosis and expansion of the dermis. CONCLUSION: Intradermal injection of processed fat results in deposition of fibrous material and collagen due to the response of the recipient site. The material itself contains very little collagen. PMID- 8245303 TI - The scalp. To lift or not to lift--is that really the question? PMID- 8245304 TI - Pooled analysis of the efficacy of bacille Calmette-Guerin (BCG) immunotherapy in malignant melanoma. AB - BACKGROUND: The trials of bacille Calmette-Guerin (BCG) as adjuvant therapy in malignant melanoma conducted over the preceding 2 decades have presented conflicting claims of efficacy. OBJECTIVE: Determination of the role of BCG immunotherapy in malignant melanoma. METHODS: Critical analysis of randomized clinical trials of stage I and II melanoma and all reported trials of intralesional and oral BCG in stage III melanoma was conducted. A literature search used the Medline data base (1966-1992);bibliographic reviews of relevant texts and pertinent articles. RESULTS: No significant benefit of BCG as postsurgical adjuvant therapy in stage I malignant melanoma was observed. Although two of seven trials in stage II melanoma demonstrated benefit with the addition of BCG, the trial with the greatest power in this series detected no difference in outcomes. In stage III malignant melanoma, there was no significant benefit with addition of BCG to various chemotherapeutic regimens. Oral BCG monotherapy produced complete responses in 6%, partial responses in 1%, and extended survival in 7% of patients. Objective responses were observed primarily in patients with intracutaneous non-visceral metastases. Pooled analysis of 15 non-controlled trials of intralesional BCG injections revealed complete responses in 19%, partial responses in 26%, and extended survival in 13% of patients with stage III melanoma. Objective responses to intralesional BCG were more likely in patients with solely cutaneous metastases, particularly intradermal lesions. CONCLUSION: Pooled analysis of non-placebo controlled trials of intralesional BCG for stage III malignant melanoma supports a trend to enhanced survival in patients with cutaneous non-visceral metastases. PMID- 8245305 TI - A comparison of copper vapor and flashlamp pumped dye lasers in the treatment of facial telangiectasia. AB - METHODS: Twelve adult patients with bilateral facial telangiectasias were treated with two yellow light lasers. A section of each involved area was treated with a copper vapor laser and a similar section in the same patient was treated with the flashlamp pumped dye laser. RESULTS: Both lasers provided satisfactory clearance at 2 and 6 weeks. Treatment was not painless with either laser; however, no patient needed local or general anesthesia. The time taken to treat equivalent areas was similar for both lasers. Postoperative swelling was greater with the flashlamp pumped dye laser than with the copper vapor laser and the time required for healing was longer with the flash-lamp pumped dye laser. CONCLUSION: The larger purpuric macules produced post-operatively by the flashlamp pumped dye laser were less cosmetically acceptable to patients when compared with the thin linear crusting produced by the copper vapor laser. Scarring or textural changes were not seen with either laser. PMID- 8245306 TI - Augmentation of in vitro cytolytic activity of LAK cells with heated ATL-derived cell lines. AB - Three adult T-cell leukemia/lymphoma-derived cell lines, MT-2, MJ, and HUT102, were investigated to determine how they responded to hyperthermia, lymphokine activated killer (LAK) cells, or a combination of both in vitro. All three cell lines showed a similar sensitivity to LAK cells, but revealed varying degrees of sensitivity to hyperthermia (MT-2 < MJ < HUT102) by 51Cr release assay. Hyperthermia did not cause immediate cell death as determined by the trypan blue exclusion test, but did cause substantial decreases in the numbers of heated cells within 2 days. The density of the cells began to increase thereafter, which was consistent with the results of the experiments labeling the cells with 3H-TdR after hyperthermia. When the cells were heated at 39-43 degrees C for 1-3 hr and then interacted with various LAK cell/ATL cell (E/T) ratios at 37 degrees C for 4 hr, total cytolysis of the cells increased in a synergistic and/or additive manner over that of the cells without hyperthermia. Prolonged incubation of the cells at high temperature did not necessarily cause a large increase in the interaction of LAK cells after hyperthermia. This augmentation of cytolysis by LAK cells after hyperthermia was not seen in normal peripheral lymphocytes. These results suggest that the combination therapy of hyperthermia and LAK cells may be more specific, useful, and effective for treating malignant lymphoma. PMID- 8245307 TI - Subungual pigmented nevus: evaluation of DNA ploidy in six cases. AB - The discrimination between subungual pigmented nevus and subungual melanoma in situ is still a clinical problem. We measured DNA ploidy in six cases of subungual melanotic lesions which exhibited the features of subungual pigmented nevus or lentigo simplex histologically. Five cases presented a diploid pattern with or without a slight increase of hyperdiploid cells. One case presented a polyploid pattern; it also exhibited histologically abnormal melanocytes with large nuclei and pigment-filled elongated dendrites. The DNA ploidy pattern and histologic features suggest that the lesion of this latter case contains abnormal melanocytes which probably have the potential to undergo a malignant transformation into a subungual melanoma. DNA ploidy analysis, therefore, is likely to provide information for evaluating the biologic behavior of subungual melanotic lesions. PMID- 8245308 TI - A case of herpes-associated erythema multiforme in a Japanese child. AB - A case of a five-year-old Japanese boy with herpes-associated erythema multiforme (HAEM) was reported. The patient had eleven recurrences of herpes labiaris within one year; four of these recurrences were accompanied by erythema multiforme. A study of the human leukocyte antigens revealed the presence of HLA-DQW3, which has been reported to be significantly frequent in Caucasian patients with HAEM. Oral administration of acyclovir at the onset of herpes labialis was effective in preventing HAEM. Early administration of oral steroids at the onset of HAEM also prevented its exacerbation. PMID- 8245309 TI - A case of verrucous carcinoma associated with human papillomavirus type 16 DNA. AB - We report here a case of verrucous carcinoma which occurred on the penis of a 75 year-old male. The nodule was first noted six months earlier and was whitish, cauliflower-like, and 17 x 19 mm in size. The histopathological examination revealed hypertrophic epidermal proliferation with pale staining keratinocytes, extending into the deep dermis. Partial penectomy and inguinal lymph node dissection were done. No lymph node metastasis was recognized. DNA was isolated from the frozen tumor tissue and examined for the presence of human papillomavirus (HPV) 16, 18, and 33 DNA by the polymerase chain reaction (PCR), using common and specific primers. A 140 base pair (bp) band was amplified and finally determined to be the HPV16 sequence by dot-blot hybridization. PMID- 8245310 TI - Linear porokeratosis. AB - A case of linear porokeratosis in a 14-year-old girl was reported. The lesions started from the right forearm and extended to the right breast. Histological examinations showed cornoid lamella in the horny layer and dyskeratotic cells in the spinous layer beneath the cornoid lamella. Linear distributions of the lesions resembled the lines of Blaschko. PMID- 8245311 TI - A case of adamantinoid basal cell epithelioma. AB - A 56-year-old woman visited the clinic of dermatology, Yokohama Sakae Kyosai Hospital in May of 1992 with a 5-year history of a blue-gray lesion on her left cheek which had enlarged asymptomatically. The tumor was excised with Z plasty closure. Histopathologically, a loosely encapsulated tumor was situated in the upper to deep dermis. At the periphery of the tumor, there were typical solid basal cell epithelioma (BCE) nests, but within the majority of lobules, intercellular spaces were widened and cells were connected by stellate appendages. Mucoid material filled the spaces and the cells at the periphery palisaded. Histochemical study revealed that the mucoid substance contained hyaluronic acid. An immunohistochemical study was performed using peroxidase antiperoxidase (PAP) method. The antibodies used were anti-epithelial membrane antigen (EMA), anti-carcinoembryonic antigen (CEA), anti-S-100 protein, anti neurofilament, anti-vimentin, anti-desmin, anti-BRST-1, and anti-BRST-2. The tumor cells and stroma expressed none of these antigens. The tumor was diagnosed as an adamantinoid BCE. PMID- 8245312 TI - The effect of cimetidine on verruca plana juvenilis: clinical trials in six patients. AB - Cimetidine is an H2-receptor antagonist which is mainly used to treat gastrointestinal diseases. Recently, many authors have suggested that cimetidine has the ability to reverse acquired tolerance to dinitrochlorobenzene (DNCB). We have used cimetidine in six patients with verruca plana who showed no therapeutic response to DNCB. Four of them responded and showed complete disappearance of the verruca plana. The mean duration of oral cimetidine administration (1.2 g/day) was 4 weeks until the disappearance. Therefore, we suggest that oral cimetidine can be used as an effective and safe adjunctive modality for the treatment of verruca plana. PMID- 8245313 TI - Clinical efficacy and side effects of acitretin on the disorders of keratinization: a one-year study. AB - Acitretin, Ro 10-1670, the principal and free acid metabolite of etretinate, was used to treat twenty patients with disorders of keratinization. An open, prospective study of clinical efficacy, tolerability, and the effects of acitretin on lipid metabolism, hepatic function, and the osteoarticular system was performed over a one year period. Each patient was initially treated with 30 mg/day of acitretin or approximately 0.6 mg/kg/day. Doses were adjusted according to the clinical efficacy and maintained for one year. There were no statistically significant changes in liver function tests or lipid profile. Twelve of eighteen evaluated patients developed asymptomatic skeletal changes; the most common change was disc space narrowing, especially at thoracic-spine level (7 of 18 patients). The earliest bone change was detected 9 months after treatment. Acitretin is effective in improving the disorders of keratinization with mild mucocutaneous side effects and asymptomatic osteoarticular changes. PMID- 8245314 TI - Cryosurgery and topical fluorouracil: a treatment method for widespread basal cell epithelioma in basal cell nevus syndrome. AB - A 58-year-old man with basal cell nevus syndrome had variously sized basal cell epitheliomas (BCEs), mostly of the superficial type, on his chest, back, and lumbar areas. BCEs on the lumbar area were treated with 5-fluorouracil (5-FU) cream which was applied daily under occlusive dressings (ODT). Complete erosion occurred in the center, but not at the periphery of the lesions. In the latter regions, BCE remained. Then cryosurgery (cryo) followed by topical 5-FU (cryo + 5 FU) was tried to treat the peripheral, non-eroded lesions; this caused complete erosions. Biopsy specimens obtained 6 months after epithelization did not show any evidence of recurrence. We also tried either cryo alone or cryo + 5-FU on the chest lesions, and either 5-FU alone or cryo + 5-FU on the abdominal lesions. Cryo alone or 5-FU alone could not clear BCE, but cryo + 5-FU could. These results suggest that the cryo + 5-FU was the most effective of these therapies. PMID- 8245315 TI - The relationship of sun protection factor to minimal erythema dose, Japanese skin type, and skin color. AB - The relationship of sun protection factor (SPF) to minimal erythema dose (MED), Japanese skin type, and skin color was investigated on the unexposed backs of 48 healthy subjects. SPF of a UVB-sunscreen was determined using sun lamps as a light source. A significant correlation was found between SPF and MED; subjects with lower MED showed higher SPF. The average SPF decreased with increasing skin type. There was no significant correlation between SPF and skin color (L*a*b* color system). PMID- 8245316 TI - Truth or consequences: dishonesty in dating and HIV/AIDS-related issues in a college-age population. AB - This article examines the issue of dishonesty in dating among college students as it relates to the HIV/AIDS epidemic in the United States. The authors surveyed 171 under-graduates at a large midwestern university in January 1991. They conducted an analysis by gender and found significant differences in responses of male and female students. Of those students involved in monogamous relationships, 36% of the men and 21% of the women reported being sexually unfaithful to their current partner or to any of their previous partners. A greater percentage of women than of men inquired about past sexual histories before engaging in sexual activity. In addition, men admitted they had lied to their sexual partner or partners more often than did women. Because a large portion of HIV/AIDS and STD education involves teaching students to inquire about previous sexual histories, the results of this study should influence the direction and content of behaviorally focused educational programs. The authors also address implications for health educators and clinicians. PMID- 8245317 TI - Research in college health. 3: Representative designs and their challenges. AB - This article explores three research designs common to college health investigations: survey, epidemiologic, and experimental. It identifies the varieties of research questions addressed by these designs and notes examples from clinical nursing/medicine, mental health, and health education topics. In addition, it summarizes the unique challenges encountered in each of these designs, including issues of sample selection, adequate measurement, and control. PMID- 8245318 TI - Six years of clinical experience using postcoital contraception in college women. AB - Postcoital contraception (PCC), also known as the "morning-after pill," has been used at the University of Rochester Health Service for several years. In 1985, the healthcare providers developed a formal protocol for dispensing PCC to female students presenting with the complaint of unprotected intercourse within the previous 72 hours. Patients are screened for any absolute contraindications to using birth control pills, are asked to sign a consent form, and are told to schedule a follow-up visit to evaluate pregnancy status and contraceptive options. Data from 1985 to 1991 is presented and include total number of times PCC was dispensed, side effects of medication, patient profiles, and predicted and actual pregnancy rates. The authors compare the data from the University of Rochester with data described in the literature and discuss recommendations for practice. PMID- 8245319 TI - Suicides among university students in Finland. AB - This study is part of a nationwide research project dealing with all suicides in Finland. It includes all suicides among university students during the 1-year period from April 1, 1987, to March 31, 1988. Ten students (6 men and 4 women) committed suicide during the period studied. The observed suicide rates per 100,000 students per year were 13.8 (95% CI = 5.1-30.1) for men and 9.0 (95% CI = 2.4-22.8) for women. The difference between the observed and expected suicide rate for men (13.8 v 47.7, standard mortality rate [SMR] = 29) was statistically significant. For women (9.0 v 8.3, SMR = 108), the difference was not statistically significant. Most of the suicides had experienced serious difficulties since early childhood, including narcissistic problems, difficulties in human relations, severe depressive periods, previous suicidal behavior, and alcohol problems; they had also had earlier psychiatric treatment. PMID- 8245320 TI - A survey of university health centers in western Canada. AB - A survey of 11 universities in the four provinces of western Canada in July 1991 found that only 1 of the institutions had no student health service. The survey asked for information about general operations, staff, finances, and services at the centers. The 10 universities that had health services were found to vary widely in their programs, funding, and compensation to physicians, with two distinct types of health service: those that serve the entire university community and others that provide care for students only. Psychiatry, preventive medicine, immunization, and sports medicine were the common services, and all centers were open throughout the year. Patient visits to the centers had increased during the preceding 5 years, although physicians working in university health services experienced approximately the same number of annual patient visits as did community physicians in private practice. The author contrasts services at the Canadian institutions with those reported by colleges and universities in the United States. PMID- 8245321 TI - College health: a model for our nation's health. The college health perspective. PMID- 8245322 TI - Correlation between college students' driving offenses and their risks for alcohol problems. AB - This study was designed to investigate the correlation between college students' driving offenses and their risk for alcohol problems. The CORE Alcohol and Drug Survey was administered to 22 undergraduate students enrolled in an alcohol and drug education program. Participants had committed an offense of driving under the influence of alcohol (DUI) or a non-DUI alcohol-related offense. The authors examined the relationship between DUI and non-DUI offenses and the following variables: age of first use, binge drinking, average number of drinks, and grade point average. The DUI group engaged in binge drinking more often and had been younger at the time of first use of alcohol than the non-DUI group. The high overall consumption and frequency of binge drinking among DUI and non-DUI offenders appears to validate alcohol and drug education programs that focus on reducing high-risk behaviors. PMID- 8245323 TI - Incidence of alcohol abuse in the parents of medical students. AB - An anonymous self-report survey of the medical students at the University of Colorado Health Sciences Center assessed how many of the students' parents were alcohol abusers. The results indicated that 27% of the student body were children of alcohol abusers, a rate twice that of the general population. This finding has implications for teaching strategies, student substance abuse, prevention efforts during the medical-school years, and provision of appropriate programs through student health services. PMID- 8245324 TI - Smoke-free universities. PMID- 8245325 TI - Culprit lesion morphology and stenosis severity in the prediction of reocclusion after coronary thrombolysis: angiographic results of the APRICOT study. Antithrombotics in the Prevention of Reocclusion in Coronary Thrombolysis. AB - OBJECTIVES: In the APRICOT study (Antithrombotics in the Prevention of Reocclusion In Coronary Thrombolysis), we sought to determine whether angiographic characteristics of the culprit lesion could predict reocclusion after successful thrombolysis and to analyze the influence of three antithrombotic treatment regimens. BACKGROUND: After successful thrombolysis, reocclusion is a major problem. Prediction of reocclusion by angiographic data and choice of antithrombotic treatment would be important for clinical management. METHODS: After thrombolysis, patients were treated with intravenous heparin until initial angiography was performed within 48 h. Patients with a patent infarct-related artery were eligible. Three hundred patients were randomly selected for treatment with coumadin, aspirin (300 mg once daily) or placebo. Patency on a second angiographic study after 3 months was the primary end point of the study. RESULTS: Reocclusion rate was 25% with aspirin, 30% with coumadin and 32% with placebo (p = NS). Lesions with > 90% stenosis reoccluded more frequently (42%) than did those with < 90% stenosis (23%) (p < 0.01). Reocclusion rate of smooth lesions was higher (34%) than that of complex lesions (23%) (p < 0.05). In lesions with < 90% stenosis, the reocclusion rate was lower with aspirin (17%) than with coumadin (25%) or placebo (30%) (p < 0.01). In complex lesions, the reocclusion rate was lower with aspirin (14%) than with coumadin (32%) or placebo (25%) (p < 0.02). Multivariate analysis showed only stenosis severity > 90% to be an independent predictor of reocclusion (odds ratio 2.31, 95% confidence interval 1.28 to 4.18, p = 0.006). CONCLUSIONS: Angiographic features of the culprit lesion after successful coronary thrombolysis significantly predict the risk of reocclusion: high grade (> 90%) stenoses reoccluded more frequently. Aspirin was effective only in complex and less severe lesions (< 90% stenosis). These findings should prompt investigation of the effects of an aggressive approach to patients with severe residual stenosis. PMID- 8245326 TI - Two- and three-year results of the Thrombolysis in Myocardial Infarction (TIMI) Phase II clinical trial. AB - OBJECTIVES: This report describes the survival and reinfarction rates for 2- and 3-year follow-up in the Thrombolysis in Myocardial Infarction (TIMI) Phase II clinical trial. BACKGROUND: Patients enrolled in TIMI II were randomly assigned to an invasive (1,681 patients) or a conservative (1,658 patients) management strategy to follow receipt of intravenous recombinant tissue-type plasminogen activator for acute myocardial infarction. METHODS: Eligibility required presentation within 4h of onset of symptoms and at least 1-mV ST segment elevation in two contiguous electrocardiographic leads. The invasive strategy group underwent cardiac catheterization 18 to 48 h after study entry and, when appropriate, percutaneous transluminal coronary angioplasty or coronary artery bypass grafting. In the conservative strategy group these diagnostic and revascularization procedures were reserved for recurrent spontaneous ischemia or ischemia on low level exercise at the time of hospital discharge. RESULTS: Complete 2-year follow-up data are available for 3,187 patients (95.4%). Cumulative life-table rates of death or reinfarction were 17.6% for the invasive strategy group and 17.9% for the conservative strategy group (p = NS) and mortality was 8.9% and 8.7% (p = NS), respectively. Complete data are available for 1,959 (90.1%) of the 2,174 patients enrolled for 3 years. Rates of death or reinfarction were 21.0% for the invasive strategy group with 20.0% for the conservative strategy group (p = NS), with mortality of 11.5% and 11.0% (p = NS), respectively. In this cohort, the mortality was 1.3% in the 2nd year and 1.7% in the 3rd year from study entry. CONCLUSIONS: TIMI II invasive and conservative strategies resulted in similar favorable outcomes after 2 and 3 years. Mortality and reinfarction rates in the two strategies were comparable. Deaths were infrequent in the 2nd and 3rd years from study entry. PMID- 8245327 TI - Incidence and significance of ventricular tachycardia and fibrillation in the absence of hypotension or heart failure in acute myocardial infarction treated with recombinant tissue-type plasminogen activator: results from the Thrombolysis in Myocardial Infarction (TIMI) Phase II trial. AB - OBJECTIVES: The purpose of this study was to determine the incidence of ventricular tachycardia and fibrillation without hypotension or heart failure after treatment with recombinant tissue-type plasminogen activator (rt-PA), anatomic correlates of their development, the effect of immediate intravenous metoprolol on their occurrence and the outcome of patients with these arrhythmias. BACKGROUND: Malignant arrhythmias after thrombolytic therapy have been reported to occur as a result of coronary reperfusion, which is associated with reduced mortality in patients receiving thrombolytic therapy. METHODS: We analyzed data from 2,546 patients in the Thrombolysis in Myocardial Infarction (TIMI) Phase II trial without congestive heart failure or hypotension during the 1st 24 h after study entry. Forty-nine patients (1.9%) developed sustained ventricular tachycardia or ventricular fibrillation within 24 h of study entry (group 1), and 2,497 patients (98.1%) did not (group 2). RESULTS: Baseline characteristics and admission laboratory values were similar in the two groups. In patients undergoing protocol angiography 18 to 48 h after rt-PA, the infarct related artery was patient in a greater percent of group 2 patients (87% [1,015 of 1,169]) than group 1 patients (68% [15 of 22], p = 0.01), although angiography was performed less frequently in group 1 than in group 2. More group 1 than group 2 patients died within 21 days (20.4%) (1.6%, p < 0.001). For patients surviving to 21 days, there was no difference in mortality between patients in the two groups in the following year. CONCLUSIONS: Ventricular tachycardia and fibrillation are not markers for reperfusion after thrombolytic therapy. These arrhythmias are associated with occlusion, not patency, of the infarct-related artery. Early mortality is increased in patients who develop ventricular tachycardia and fibrillation, even in the absence of congestive heart failure and hypotension. PMID- 8245328 TI - Thrombolytic therapy for women with myocardial infarction: is there a gender gap? Thrombolysis and Angioplasty in Myocardial Infarction Study Group. AB - OBJECTIVES: The goal of this study was to investigate whether female gender portends an adverse prognosis independent of the severity of the underlying disease after acute myocardial infarction treated by thrombolysis. A total of 348 women were compared with 1,271 men enrolled in the Thrombolysis and Angioplasty in Myocardial Infarction (TAMI) trials. BACKGROUND: The reasons for gender differences in the management and prognosis of acute coronary artery syndromes remain poorly defined. The extent to which gender itself explains observed differences in outcome and use of diagnostic procedures remains unclear because confounding factors have not been specified. METHODS: Patients < 76 years of age presenting within 6 h of onset of ischemic symptoms with electrocardiographic ST segment elevation and without contraindications to thrombolysis, previous infarction in the same distribution or cardiogenic shock were prospectively enrolled in Phases 1 to 3, 5 and 7 of the TAMI trials. All patients received recombinant tissue-type plasminogen activator, urokinase or a combination of both agents. Protocol-mandated cardiac catheterization was performed during the hospital period. Rescue coronary angioplasty was carried out for reperfusion failure at angiography 90 min after initiation of thrombolytic therapy. Coronary artery bypass grafting or coronary angioplasty was performed for clinical indications. RESULTS: Women were older than men (61.0 +/- 9.7 vs. 55.8 +/- 10.1 years, mean +/- SD) and had a higher incidence of many risk factors for adverse outcome after myocardial infarction. There were no differences in baseline hemodynamic variables or time to thrombolytic treatment. Rates of acute and predischarge infarct-related artery patency and global and regional left ventricular function were similar in the two groups. Rates of in-hospital coronary angioplasty (52.6% and 54.1%) and bypass graft surgery (20.4% and 22.0%) were comparable in women and men, respectively. Women had higher unadjusted rates of mortality (9.2% vs. 5.4%, p = 0.014), reinfarction (6.4% vs. 2.6%, p = 0.005) and hemorrhagic stroke (2.0% vs. 0.55%, p = 0.017) than did men during the hospital period. When adjusted for clinical and angiographic variables, differences in mortality and hemorrhagic stroke did not reach statistical significance, and the risk of reinfarction was only marginally associated with gender. CONCLUSIONS: In selected patients undergoing thrombolytic therapy and cardiac catheterization for acute myocardial infarction, adjusted mortality rates and utilization of postlysis revascularization are similar in women and men. However, women may be at increased risk for reinfarction. PMID- 8245329 TI - Influence of diabetes on mortality in acute myocardial infarction: data from the GISSI-2 study. AB - OBJECTIVES: This study was conducted to determine the role of insulin-dependent and noninsulin-dependent diabetes in the prognosis of patients after myocardial infarction and treatment with fibrinolytic agents. BACKGROUND: Several studies have shown that diabetic patients have a high mortality rate after acute myocardial infarction. However, the impact of diabetes on survival in patients treated with fibrinolytic agents is still undefined. It is also not known whether the type of diabetes or gender affects prognosis. METHODS: We analyzed prevalence and prognostic significance of a history of diabetes in patients enrolled in the GISSI-2 study, all of whom received fibrinolytic agents. The incidence of deaths in the hospital and at 6 months after study entry was computed for patients without diabetes and for insulin-dependent and noninsulin-dependent diabetic patients; relative risks were evaluated by univariate and multivariate analysis. RESULTS: Information on diabetic status was available for 11,667 patients, 94.2% of those randomized in the GISSI-2 study. The prevalence of diabetes was higher in women than in men (8.75% vs. 1.85%, p < 0.01 for insulin-dependent and 23.7% vs. 13.8%, p < 0.01 for noninsulin-dependent diabetic patients). The type of fibrinolytic agent did not affect mortality rates; the increase in in-hospital mortality of diabetic patients was moderate and similar for men with insulin- and noninsulin-dependent diabetes (8.7% and 10.1%, respectively, vs. 5.8% in nondiabetic patients); in women, mortality was markedly higher for insulin dependent and only slightly higher for noninsulin-dependent diabetic patients (24.0% and 15.8%, respectively, vs. 13.9% for nondiabetic patients). The adjusted relative risks were 1.9 (95% confidence interval 1.2 to 2.9) for insulin dependent diabetic women and 1.4 (95% confidence interval 1.1 to 1.8) for noninsulin-dependent diabetic men. The mortality rate after discharge showed a similar gender difference, and in insulin-dependent diabetic women, prognosis was ominous even in the absence of left ventricular damage before discharge. CONCLUSIONS: A history of diabetes is associated with a worse prognosis after myocardial infarction, even in patients treated with fibrinolytic agents. Gender and type of diabetes appear to be critical in affecting survival. In men, both insulin-dependent and noninsulin-dependent diabetes are associated with a moderately higher mortality rate; in women, insulin-dependent diabetes is, in itself, a strong risk factor for death after myocardial infarction. PMID- 8245330 TI - Feasibility of early discharge after acute Q wave myocardial infarction in patients not receiving thrombolytic treatment. AB - OBJECTIVES: The purpose of this study was to analyze the feasibility of early discharge (4 days) after acute myocardial infarction in patients not receiving thrombolytic therapy by first identifying predictors of short-term prognosis and then testing the derived risk profile in an independent cohort of patients. BACKGROUND: Previous studies have shown that early discharge after acute myocardial infarction is possible. However, physicians are reluctant to shorten the standard 7- to 10-day hospital stay, presumably because of difficulty in selecting low risk patients. METHODS: From January 1985 to November 1986, 358 patients with acute myocardial infarction who did not receive thrombolytic therapy were screened. Those with a Q-wave infarction showing no complications on day 4 were considered candidates for early discharge and were transferred to the ward for a mean of 12 days. During this period, we looked for any event (cardiac or noncardiac) that would have prompted readmission if the patient had been previously discharged. Univariate and multiple regression analysis were performed to identify predictors of these events among 25 baseline variables. The derived risk profile was tested in an independent validation cohort. RESULTS: One hundred five (29.3%) of the 358 patients were free of symptoms on day 4, and 29 (27.6%) had at least one cardiac event, including four deaths and one reinfarction. Multivariate analysis selected diabetes, ejection fraction < 40% and age as independent predictors of events. Using the risk profile, 18 (13.2%) of the 136 validation cohort patients were categorized as low risk, and only 1 of them had a major event (progressive angina). Sensitivity for the risk profile was high (91%), but specificity was low (34%). CONCLUSIONS: The use of simple clinical variables may allow the safe reduction of hospital stay after infarction in selected patients. However because the proportion of candidates for early discharge is small (12.6%), it seems unlikely that the current policies on length of hospital stay will change in the near future. PMID- 8245331 TI - Early discharge after a myocardial infarction: what's the hurry? PMID- 8245332 TI - Emergency room technetium-99m sestamibi imaging to rule out acute myocardial ischemic events in patients with nondiagnostic electrocardiograms. AB - OBJECTIVES: The goal of this study was to determine the role of nuclear imaging in patients with chest pain. BACKGROUND: The diagnosis of myocardial ischemic events in patients with chest pain and a nondiagnostic electrocardiogram (ECG) is problematic. METHODS: Rest tomographic technetium-99m sestamibi imaging (740 MBq intravenously) was performed in 64 patients presenting to the emergency room with chest pain of suspected cardiac origin and a nondiagnostic ECG. Patients were admitted to the coronary care unit on the basis of clinical criteria only and were strictly monitored. RESULTS: Thirty patients showed a perfusion defect on admission. Of these, 13 developed myocardial infarction within 12 h. Coronary artery disease was diagnosed in 14 patients and the remaining 3 patients were classified as having false positive findings. Normal perfusion scans were seen in 34 patients, none of whom were ultimately diagnosed as having coronary artery disease. A 100% sensitivity was demonstrated versus the final diagnosis of acute cardiac ischemia (kappa 0.91, 95% confidence interval 0.8 to 1.0). A follow-up period of up to 18 months (mean 11 +/- 3) was also carried out for major cardiac events (death, myocardial infarction, coronary angioplasty and coronary artery bypass grafting). Six events (two coronary bypass procedures, three angioplasty procedures and one death) were observed at follow-up in the group of patients with a technetium-99m sestamibi perfusion defect. Patients with normal perfusion scans on admission had no major cardiac events at follow-up study. CONCLUSIONS: Technetium-99m sestamibi perfusion imaging is a promising technique for ruling out acute myocardial ischemia in the emergency room. More efficient utilization of intensive therapy beds may be expected with this approach. PMID- 8245333 TI - Prognostic value of dipyridamole echocardiography early after myocardial infarction in elderly patients. Echo Persantine Italian Cooperative (EPIC) Study Group. AB - OBJECTIVES: This study was conducted to assess the feasibility, safety and prognostic value of dipyridamole echocardiography in elderly patients recovering from an uncomplicated acute myocardial infarction in a subset analysis performed on the patients entered in the subproject "residual ischemia" of the Echo Persantine Italian Cooperative Study (EPIC). BACKGROUND: Coronary heart disease accounts for two thirds of all deaths in the age group > 65 years, and > 50% of all patients admitted to the hospital with acute myocardial infarction are > 65 years old. The prognostic value of dipyridamole-induced left ventricular dysfunction was clearly established in patients evaluated early after acute infarction. METHODS: In a subgroup analysis of the Echo Persantine Italian Cooperative Study (EPIC), we assessed the value of dipyridamole echocardiography in predicting cardiac events in 190 elderly (> or = 65 years) patients (age 68.4 +/- 3.3 years, range 65 to 78; 147 men and 43 women) evaluated early (mean 10 days) after uncomplicated acute myocardial infarction and followed up for 14 +/- 9.8 months. RESULTS: There was no major side effect during dipyridamole echocardiography. A positive test result occurred in 85 patients (44.7%). During follow-up, there were 62 events (14 cardiac deaths, 7 nonfatal reinfarctions, 21 cases of class III or IV angina and 20 revascularization procedures). Of these 62 events, 44 occurred among 85 patients with positive dipyridamole echocardiography and 18 among 105 patients with negative dipyridamole echocardiography (52% vs. 17%, p < 0.001). Spontaneous events (death, reinfarction, angina) occurred in 31 patients with positive and in 11 with negative dipyridamole echocardiography (36% vs. 10%, p < 0.001). Hard events (myocardial infarction or death) occurred in 14 patients with positive and 7 with negative dipyridamole echocardiography (16% vs. 6%, p < 0.05). Death occurred in 11 patients with positive and in 3 with negative dipyridamole echocardiography (13% vs. 3%, p < 0.01). The positive predictive value of positive dipyridamole echocardiography and negative predictive value of negative dipyridamole echocardiography as related to the occurrence of all events in the follow-up period (death, reinfarction, angina, revascularization procedures) were 52% and 83%, respectively. The relative risk (that is, the relative risk of occurrence of future cardiac events in the group with positive dipyridamole echocardiography compared with that in those with negative dipyridamole echocardiography) was 3 for all events and 4.4 for death. CONCLUSIONS: Dipyridamole echocardiography was well tolerated by elderly patients and proved to be very effective in prognostic stratification early after uncomplicated acute myocardial infarction, even when only survival was considered. PMID- 8245334 TI - Comparison of symptom-limited and low level exercise tolerance tests early after myocardial infarction. AB - OBJECTIVES: This study was conducted to determine the diagnostic yield and risks of a symptom-limited treadmill exercise test before hospital discharge. BACKGROUND: Currently, predischarge low level and 6-week symptom-limited exercise treadmill tests are recommended for risk stratification after myocardial infarction. However, few data exist on the safety and value of a predischarge symptom-limited exercise test. METHODS: We utilized a modified Bruce protocol starting at 1.7 mph and 0 grade with 3-min stages in 150 consecutive patients 6.4 +/- 3.1 days after myocardial infarction. Each exercise test was interpreted for duration, symptoms and ST segment changes at the low level (70% of predicted heart rate) and symptom-limited end point. RESULTS: There were no complications related to the symptom-limited exercise tests. The test results were positive in only 23% of the patients at the low level end point, but were positive in 40% of the patients at the later symptom-limited end point (p < 0.001). During a mean follow-up period of 15 +/- 5 months in 138 patients (92%), 50 patients (36%) had a cardiac event. Of the patients with a cardiac event, significantly more (p < 0.001) had a positive exercise test at the symptom-limited end point (31 vs. 16 patients). Five patients with a negative and 14 patients with a nondiagnostic symptom-limited exercise test had an event. CONCLUSIONS: In patients with uncomplicated myocardial infarction, we demonstrated the safety of an early symptom-limited treadmill exercise test. Symptom-limited exercise tests will identify more patients with inducible ischemia who are at risk of future cardiac events and who may benefit from early intervention. PMID- 8245335 TI - Long-term physical training and left ventricular remodeling after anterior myocardial infarction: results of the Exercise in Anterior Myocardial Infarction (EAMI) trial. EAMI Study Group. AB - OBJECTIVES: The aim of this multicenter randomized study was to investigate whether long-term physical training would influence left ventricular remodeling after anterior myocardial infarction. BACKGROUND: Exercise is currently recommended for patients after myocardial infarction; however, the effects of long-term physical training on ventricular size and remodeling still have to be defined. METHODS: Patients with no contraindications to exercise were studied 4 to 8 weeks after anterior Q wave myocardial infarction and 6 months later by echocardiography at rest and bicycle ergometric testing. After the initial study, patients were randomly allocated to a 6-month exercise training program (n = 49) or a control group (n = 46). A computerized system was used to derive echocardiographic variables of ventricular size, function and topography. RESULTS: After 6 months, a significant (p < 0.01) increase in work capacity (from 4,596 +/- 1,246 to 5,508 +/- 1,335 kp-m) was observed only in the training group, whereas global ventricular size, regional dilation and shape distortion did not change in either the control or the training group. However, compared with patients with an ejection fraction > 40%, patients with an ejection fraction < or = 40% had more significant (p < 0.001) ventricular enlargement at entry and demonstrated further (p < 0.01) global and regional dilation after 6 months, in both the control and the training group (end-diastolic volume from 77 +/- 14 to 85 +/- 17 ml/m2 in the control group and from 74 +/- 11 to 77 +/- 15 ml/m2 in the training group; regional dilation from 46 +/- 18% to 57 +/- 21% in the control group and from 42 +/- 18% to 44 +/- 26% in the training group). Ventricular size and topography did not change in patients with an ejection fraction > 40%. CONCLUSIONS: Patients with poor left ventricular function 1 to 2 months after anterior myocardial infarction are prone to further global and regional dilation. Exercise training does not appear to influence this spontaneous deterioration. Thus, postinfarction patients without clinical complications, even those with a large anterior infarction, may benefit from long-term physical training without any additional negative effect on ventricular size and topography. PMID- 8245336 TI - Determinants of successful nonthoracotomy cardioverter-defibrillator implantation: experience in 101 patients using two different lead systems. AB - OBJECTIVES: This study was conducted to identify the determinants of successful nonthoracotomy cardioverter-defibrillator implantation. BACKGROUND: Until recently, either median sternotomy or thoracotomy was necessary to implant the electrodes used for internal cardioverter-defibrillator systems. A number of manufacturers have developed nonthoracotomy lead systems comprising two transvenous coil electrodes and a subcutaneous patch electrode. At present, the factors associated with the success or failure of a nonthoracotomy approach are unknown. METHODS: A total of 101 consecutive patients requiring a cardioverter defibrillator underwent an initial nonthoracotomy approach. Factors associated with successful nonthoracotomy implantation were prospectively determined. RESULTS: A nonthoracotomy system was implanted in 72 (71%) of 101 patients. Twenty-nine patients (29%) required thoracotomy. Univariate predictors of successful nonthoracotomy implantation included smaller cardiac size (p < 0.0001), smaller cardiothoracic ratio (p < 0.0002), QRS duration < 120 ms (p = 0.003), female gender (p = 0.006), ventricular fibrillation as the presenting arrhythmia (p = 0.03) and smaller echocardiographic left ventricular size (p = 0.04). Multivariate predictors included smaller cardiac size (p < 0.002) and female gender (p < 0.007). Total actuarial survival over a mean (+/- SD) follow up interval of 12 +/- 7 months was 91 +/- 0.03% and was not different in the thoracotomy and nonthoracotomy groups. CONCLUSIONS: A nonthoracotomy cardioverter defibrillator system can be implanted in a majority of patients. Smaller cardiac size and female gender are associated with a high probability of successful implantation. PMID- 8245337 TI - A placebo-controlled trial of intravenous and oral disopyramide for prevention of neurally mediated syncope induced by head-up tilt. AB - OBJECTIVES: A double-blind randomized trial was designed to determine the efficacy of intravenous and oral disopyramide phosphate in preventing neurally mediated syncope induced by a head-up tilt test. BACKGROUND: Neurally mediated syncope is a frequent cause of syncope and may be induced by head-up tilt testing. Recent uncontrolled trials have suggested that disopyramide may be an effective therapy in patients with neurally mediated syncope. METHODS: Twenty-two consecutive patients with recurrent neurally mediated syncope and two or more successive positive head-up tilt test responses were randomly allocated to receive either intravenous disopyramide or placebo. Head-up tilt testing at 60 degrees was performed for 15 min. If presyncope or syncope was not provoked, isoproterenol infusion was started at a rate of 1 microgram/min and the rate gradually increased until a 25% increase in heart rate was achieved. Eleven patients were subsequently randomized in crossover fashion to receive oral disopyramide (800 mg/day) or placebo during 1 week. The primary end point was prevention of syncope or presyncope provoked by head-up tilt testing. RESULTS: Head-up tilt test results were positive for syncope in 12 (75%) of 16 patients receiving intravenous placebo and in 12 (60%) of 20 patients receiving disopyramide (p = 0.55 Fisher exact test, 95% confidence interval [CI] -14% to 40%). In the intravenous phase, complete crossover was achieved in 15 patients. Head-up tilt test results during this phase were positive in 13 patients (87%) receiving placebo and in 12 patients (80%) receiving disopyramide (p = 0.50 Fisher exact test, 95% CI -19% to 32%) and were positive in all patients receiving their initially randomized drug or placebo. In the oral phase, head-up tilt results were positive in only two patients (18%) assigned to placebo and in three patients (27%) receiving disopyramide (p = 0.54 Fisher exact test, 95% CI 42% to 24%). A mean follow-up time of 29 +/- 8 months was obtained in 21 of the 22 patients. Syncope recurred in 3 (27%) of the 11 patients receiving disopyramide and 3 (30%) of the 10 patients not treated pharmacologically (p > 0.05). CONCLUSIONS: Intravenous disopyramide was ineffective for the prevention of neurally mediated syncope provoked by head-up tilt testing. No significant effect was observed after oral therapy with disopyramide. There was a striking decrease in the incidence of positive tilt test results over time regardless of intervention, thus discouraging the use of head-up tilt as the single method of assessing therapeutic efficacy. Recurrence of syncope after the investigative protocol was infrequent over long-term follow-up regardless of treatment group. PMID- 8245338 TI - Elimination of slow pathway conduction: an accurate indicator of clinical success after radiofrequency atrioventricular node modification. AB - OBJECTIVES: The purpose of this study was to determine the optimal end point of radiofrequency atrioventricular (AV) node modification using anatomically guided slow pathway approaches in patients with AV node reentrant tachycardia. BACKGROUND: The optimal end point for AV node modification using radiofrequency energy is uncertain, although elimination of inducible AV node reentrant tachycardia has been used. METHODS: We followed up 51 consecutive patients (40 women, 11 men, mean age +/- SD 41 +/- 16 years) with symptomatic AV node reentrant tachycardia for 12 +/- 6 months (range 4 to 24) after radiofrequency AV node modification using an anatomically guided slow pathway approach. Inducible AV node reentrant tachycardia was eliminated in all patients, whereas residual slow pathway conduction persisted in 12 patients (24%) after ablation. One study was complicated by complete AV block and two patients were lost to follow-up (one with and one without residual slow pathway conduction). RESULTS: Clinical recurrence of AV node reentrant tachycardia was documented in seven patients (14%) 3 days to 3 months (median 1 month) after ablation. The recurrence rate was significantly higher in patients with than in those without residual slow pathway conduction (6 [55%] of 11 vs. 1 [3%] of 37, p < 0.01). The recurrence rate was not different between patients with only residual slow pathway conduction and those with residual slow pathway conduction and inducible single echo cycles (three [60%] of five in both groups, p = NS). The number of radiofrequency energy applications was not significantly different between those without and those with recurrence (20 +/- 17 vs. 16 +/- 9, p = NS). Junctional tachycardia during application of radiofrequency energy tended to be more frequently observed in those with a successful outcome (77% vs. 57%, p > 0.05). Of the 22 patients who underwent modification before 1992, residual slow pathway conduction was present in 9 (41%) of 22 patients. Atrioventricular node reentrant tachycardia recurred in five (56%) of these nine patients. A greater effort made in 1992 to eliminate slow pathway conduction in 29 patients resulted in residual slow pathway conduction in only 3 (11%) with recurrence in 2 (4%). CONCLUSIONS: Complete elimination of slow pathway conduction is feasible in the majority of patients. Elimination of slow pathway conduction is highly predictive of long-term success after AV node modification using an anatomically guided approach. PMID- 8245339 TI - Atrial sensing performance of the single-lead VDD pacemaker during exercise. AB - OBJECTIVES: The purpose of this study was to examine the atrial sensing performance of the single-lead VDD pacing system during exercise and concomitant changes in the amplitude of the atrial electrogram. BACKGROUND: Studies of conventional dual-chamber pacing have demonstrated an overall reduction in the atrial signal amplitude and a variable incidence of atrial undersensing during vigorous exercise. METHODS: The telemetered atrial electrogram and simultaneous surface electrocardiogram (ECG) were continuously recorded in 12 patients (mean age 70.8 years) with an implanted single-lead VDD pacing system during treadmill stress testing. The atrial signal amplitude was measured at peak exercise, and the patients were monitored for maintained atrial synchronized ventricular pacing during the entire exercise and recovery period. RESULTS: The atrial electrographic voltage decreased an average of 19.5% (p < 0.05) during peak exercise, and the reduced P wave amplitude ranged from 6.9% to 59.4% of the preexercise values in 8 of 12 patients. Three patients showed a modest increase in atrial signal amplitude, and one patient had no change. The telemetered electrogram displayed persistent and intact atrial synchronous ventricular pacing throughout the study period. CONCLUSIONS: Despite relatively low atrial signal amplitudes at rest and further decreases during exercise, the single-lead VDD pacemaker maintains reliable atrial tracking and ventricular pacing during vigorous exercise. PMID- 8245340 TI - Intravascular ultrasound study of angiographically mildly diseased coronary arteries. AB - OBJECTIVES: We hypothesized that intravascular ultrasound may identify significant coronary artery narrowing in the mildly diseased coronary artery of patients with insignificant or one- or two-vessel coronary artery disease. BACKGROUND: Necropsy studies have revealed that coronary angiography may underestimate stenosis severity in vessels that appear mildly diseased. Intravascular ultrasound has been shown to detect atherosclerotic changes in angiographically normal coronary arteries and to correlate better with histologic findings. METHODS: In 20 patients, we performed intravascular ultrasound imaging (3.5F catheter, 30-MHz transducer) in 37 coronary arteries that were considered mildly diseased (<50% diameter narrowing) by qualitative angiography. The angiographic diagnosis was no significant coronary artery disease in eight patients, one-vessel disease in seven and two-vessel disease in five. Each vessel, except for the left main coronary artery, was divided into proximal, mid and distal segments. Percent area narrowing and minimal lumen diameter were subsequently quantified by both ultrasound and quantitative angiography. RESULTS: Mean maximal arterial area narrowing by ultrasound in the 67 segments studied was 36 +/- 20% (range 0% to 80.2%) and 19 +/- 23% (range 0% to 82%) by quantitative angiography of these same segments (p < 0.001, paired t test). Mean minimal lumen diameter of the segment was 3.3 +/- 0.9 mm by ultrasound and 2.7 +/- 0.8 mm by quantitative angiography. In 10 patients there were 19 angiographically mildly diseased segments where the percent arterial area narrowing by ultrasound was > or = 50%. Intravascular ultrasound revealed that the more proximal (reference) segment had > 25% intimal thickening in 12 of the 19 underestimated segments. In six stenosed segments (32%), total vessel area increased compared with that of the adjacent proximal vessel segment because of compensatory dilation. CONCLUSIONS: Intravascular ultrasound identifies potentially significant coronary artery disease in vessels that appear to be only mildly diseased by angiography. PMID- 8245341 TI - Impairment of the arterial baroreflex during symptomatic and silent myocardial ischemia in humans. AB - OBJECTIVES: The aim of this study was to assess whether transient episodes of symptomatic or silent myocardial ischemia after baroreceptor modulation of heart rate. BACKGROUND: Animal and human studies have shown that myocardial infarction is accompanied by an impairment of the baroreceptor influences on the sinus node. However, whether this also occurs during transient myocardial ischemia has never been documented. METHODS: In 12 patients undergoing coronary angiography, systolic blood pressure (intraarterial catheter) was reduced by an intravenous bolus of nitroglycerin during a spontaneous episode of transient chest pain and myocardial ischemia (ST segment depression on the electrocardiogram) and 30 min after recovery. The slope of the linear regression between the decrease in systolic blood pressure and the RR interval shortening was taken as the measure of baroreflex sensitivity. RESULTS: During ischemia, baroreflex sensitivity was 1.3 +/- 0.3 ms/mm Hg (mean +/- SEM), whereas after recovery it was markedly and significantly greater (2.6 +/- 0.5 ms/mm Hg, p < 0.01). Similar results were obtained in eight other patients who experienced a silent ischemic episode either spontaneously or during coronary angioplasty. The reduction in baroreflex sensitivity was similarly pronounced during inferior (10 patients) and anterior (10 patients) ischemia, and its magnitude showed little or no relation to the ischemia-dependent changes in blood pressure and heart rate. CONCLUSIONS: Transient myocardial ischemia is associated with marked baroreflex impairment. The impairment occurs even during symptomless ischemic episodes and is therefore not related to pain or to other nonspecific influences on the baroreflex. PMID- 8245342 TI - Relation of arterial pressure waveform to left ventricular and carotid anatomy in normotensive subjects. AB - OBJECTIVES: The purpose of this study was to examine the relation of the central arterial pressure waveform to left ventricular and carotid structure. BACKGROUND: The pressure waveform in the central arteries is affected by reflection of the pressure wave from the periphery. When reflected waves merge with the incident wave during systole, a late systolic peak and increment in systolic blood pressure are observed. The consequent increase in hemodynamic load may stimulate left ventricular and vascular adaptive changes. METHODS: Sixty-seven normotensive adults were studied by noninvasive techniques. Anatomy and function of the left ventricle and carotid artery were investigated by ultrasonography. Pressure waveforms were recorded by an external tonometer applied to the carotid artery, and waveform shape was expressed by the augmentation index, calculated from the difference between the maximal systolic pressure and that at the inflection between early and late systolic pressure peaks divided by the pulse pressure. Subjects were assigned to groups with a dominant early (group 1, augmentation index < or = 0) or dominant late systolic peak (group 2, augmentation index > 0). RESULTS: Left ventricular mass index was significantly higher in group 2 than in group 1, a difference that persisted after controlling for the confounding effects of gender, age and blood pressure. Carotid wall thickness and regional arterial stiffness were significantly increased in group 2, but differences disappeared in the analysis of covariance for age. CONCLUSIONS: Left ventricular and carotid artery structure are related to the shape of the central pressure waveform. Although the increase in left ventricular mass seen in subjects with a dominant late systolic peak pressure appears to be directly related to the shape of the pressure waveform, changes in the structural and physical properties of the carotid artery appear to be more closely related to the aging process. PMID- 8245343 TI - Short- and long-term effects of smoking on arterial wall properties in habitual smokers. AB - OBJECTIVES: This study investigated the short-term effects of smoking on hemodynamic function and distensibility and compliance of large arteries in habitual smokers. In addition, the effect of smoking was not measured in nonsmokers, but vessel wall properties were compared between smokers and nonsmokers (basal state). BACKGROUND: Smoking is a well known risk factor for atherosclerosis. Loss of distensibility and compliance of large arteries may play a role in the onset of atherosclerosis. METHODS: The distensibility and compliance coefficients of the common carotid and brachial arteries were determined from the arterial wall displacement during systole and the end diastolic diameter by using a vessel wall movement detector and from the pulse pressure as assessed in the upper arm. Cardiac function (cardiac output, stroke volume) was measured with Doppler echocardiography. Systemic vascular resistance was calculated as mean arterial pressure divided by cardiac output. RESULTS: In habitual smokers, smoking one cigarette caused a sharp increase in blood pressure (6%) and heart rate (14%). Cardiac index increased (16%), mainly because of the marked increase in heart rate. Stroke and systemic vascular resistance indexes did not change significantly. Smoking enhanced forearm blood flow after wrist occlusion (17%), but total forearm blood flow was unchanged, suggesting an increase in muscle blood flow and a decrease in skin flow. Because of higher blood pressure, the diameter of the elastic common carotid artery increased by 3% (passive phenomenon). Distensibility of the carotid artery decreased (7%), and as a result, carotid compliance was preserved. In contrast, despite higher blood pressure, the diameter of the muscular brachial artery did not change, suggesting an increased vascular tone. Brachial distensibility and compliance decreased (18% and 19%, respectively). Habitual smokers were comparable to nonsmokers with regard to blood pressure, cardiac function, vascular resistance and vessel wall properties of large arteries. Heart rate was higher in habitual smokers (14%). CONCLUSIONS: These data indicate that in habitual smokers, smoking one cigarette causes short-term increases in arterial wall stiffness that might be harmful to the artery and increase the risk for plaque rupture. Except for a higher heart rate, no obvious long-term effect of smoking was observed on hemodynamic variables and arterial stiffness. Because acute cardiovascular events are mainly due to plaque rupture, the short-term effects of smoking might be a more important risk than long-term effects for these acute ischemic events. PMID- 8245344 TI - A novel strategy for stent deployment in the treatment of acute or threatened closure complicating balloon coronary angioplasty. Use of short or standard (or both) single or multiple Palmaz-Schatz stents. AB - OBJECTIVES: The purpose of this study was to determine the immediate and long term angiographic and clinical results of coronary stenting as a specific therapy for intracoronary dissection associated with acute or threatened closure complicating percutaneous transluminal coronary angioplasty. BACKGROUND: Published reports contain conflicting results with regard to the benefit of stent insertion for coronary dissection. In particular, there is a wide range in the reported rates of subacute occlusion. METHODS: Palmaz-Schatz stents were inserted in 56 patients who had significant dissections and acute or threatened closure complicating coronary angioplasty. An attempt was made to cover the entire site of the dissection with short or standard single or multiple Palmaz-Schatz stents. The use of the short stent allowed complete coverage of the dissection, specifically in situations such as marked vessel tortuosity or the need to place a stent distal to a deployed stent. RESULTS: A single stent was implanted in 24 patients and multiple stents were implanted in 32 patients. A total of 138 stents (78 standard, 60 short stents) were implanted. The primary clinical success rate was 88% (49 of 56 patients). Complications occurred in seven patients (12.5%): Three patients (5%) required urgent bypass surgery; two patients (4%) had a myocardial infarction; and two patients (4%) died. Subacute occlusion occurred in one patient (2%). Clinical follow-up was available in all patients at a mean of 10 +/- 4 months. Thirty-nine (80%) of 49 patients were clinically asymptomatic. Angiographic restenosis was found in 15 (36%) of 42 patients on angiographic follow-up performed a mean of 5 months (median 6) after the procedure in 86% of the eligible patients. Nine patients had successful repeat angioplasty, and two had elective bypass surgery. CONCLUSIONS: The strategy of coronary stenting to completely cover the lesion is an effective treatment for large coronary dissection complicating angioplasty. A total major complication rate of 12.5% may be acceptable for this high risk group. PMID- 8245345 TI - Mechanisms of cardiac pain during coronary angioplasty. AB - OBJECTIVES: This study was conducted to establish whether the cardiac pain patients experience during coronary angioplasty is modulated by 1) the stretching of the coronary artery wall, and 2) the mechanisms responsible for the ischemic preconditioning. BACKGROUND: Anecdotal experimental observations indicate that stretching of the coronary artery wall is a stimulus adequate to cause cardiac pain. Furthermore, recent experimental studies indicate that adenosine, a mediator of the anginal pain, appears to play an important role in the genesis of ischemic preconditioning. METHODS: We randomly allocated 48 consecutive patients undergoing coronary angioplasty into two groups. In Group A the second balloon inflation was performed at a higher level than the first; in Group B the first two inflations were performed at the same level of balloon pressure. The mean values (+/- 1 SD) of ST segment shift on the surface 12-lead electrocardiogram (ECG) and the intracoronary ECG were measured at the end of each inflation period. Severity of cardiac pain was also obtained at the same time by using a visual analog scale. RESULTS: The mean ST segment shift during the second balloon inflation was significantly less than that during the first inflation in both groups of patients (12.8 +/- 9.3 vs. 18.5 +/- 11.9 mm, p < 0.001 and 13.7 +/- 10.1 vs. 21.3 +/- 13.9 mm, p < 0.001, respectively, in Groups A and B). Yet, the severity of cardiac pain during the second inflation was greater than that during the first inflation in Group A (40.8 +/- 32.7 vs. 26.9 +/- 27.2 mm, p < 0.01), whereas it was lesser in Group B (23.1 +/- 20.7 vs. 32.9 +/- 29.6 mm, p < 0.05). However, in the latter group, pain severity after normalization for the mean ST segment shift was similar during the first and second inflations (2.1 +/- 2.4 vs. 2.7 +/- 3.6, p = NS). CONCLUSIONS: During coronary angioplasty, the cardiac pain experienced by patients is caused in part by stretching of the coronary artery wall. If the stretching is maintained at a constant level during repeated coronary occlusions, the cardiac pain is entirely predicted by the severity of myocardial ischemia and therefore does not appear to be directly modulated by the mechanisms responsible for the ischemic preconditioning. PMID- 8245346 TI - Increased levels of serum neopterin and decreased production of neutrophil superoxide anions in chronic heart failure with elevated levels of tumor necrosis factor-alpha. AB - OBJECTIVES: The purpose of this study was to examine the role of tumor necrosis factor-alpha and tetrahydrobiopterin and superoxide anion release from neutrophils in severe chronic heart failure. BACKGROUND: Previous studies have demonstrated elevated production of tumor necrosis factor-alpha and free radical induced endothelial cell damage in severe heart failure. METHODS: Plasma and serum levels of immunoreactive interleukin-1, interleukin-6, interferon-gamma, neopterin and tumor necrosis factor-alpha and the release of superoxide anions from circulating neutrophils both at basal conditions and after triggering with f Met-Leu-Phe or phorbol 12-myristate 13-acetate were measured in 16 patients with severe heart failure and in 11 healthy control subjects. RESULTS: Circulating levels of tumor necrosis factor-alpha and neopterin were elevated in patients with heart failure compared with values in control subjects. A significant correlation between the two was found. Basal and phorbolester-triggered release of oxygen radicals from neutrophils was not affected in patients with heart failure. However, formylpeptide-stimulated release of oxygen radicals by neutrophils was significantly reduced. CONCLUSIONS: Suppressed neutrophil function in patients with heart failure exhibiting elevated levels of tumor necrosis-alpha factor may indicate self-protection against the deleterious effects of neutrophil-derived oxygen radicals. Through induction of tetrahydrobiopterin synthesis (as reflected by increased neopterin), tumor necrosis factor-alpha may affect nitric oxide synthesis. PMID- 8245347 TI - Up-regulation of beta 2-adrenergic receptors in previously transplanted, denervated nonfailing human hearts. AB - OBJECTIVES: The purpose of this study was to examine beta-adrenergic receptor signal transduction in denervated, previously transplanted human ventricular myocardium. BACKGROUND: In model systems, surgical denervation typically results in both presynaptic and postsynaptic supersensitivity in beta-adrenergic receptor pathways and alteration in G protein-mediated signal transduction. METHODS: We examined beta-adrenergic receptor signal transduction in the left and right ventricles removed from nine subjects with a previous transplant and surgical denervation 25 +/- 4 months after their first transplantation. Twenty-six hearts removed from organ donors served as control hearts. RESULTS: Total beta adrenergic receptor density and stimulation of muscle contraction in isolated right ventricular trabeculae by the nonselective agonist isoproterenol were similar in the transplant and donor groups. Beta 1-receptor density was not different in the left ventricles of the two groups but tended to be reduced (by 29%, p = 0.09) in transplant right ventricles. By contrast, beta 2-receptor density was higher in transplant left and right ventricles relative to the respective values in donor ventricles by 33% in left ventricles and 97% in right ventricles (both p < 0.05). Isoproterenol, which in particulate fractions of human heart stimulates adenylyl cyclase primarily via beta 2-receptors, produced a greater increase in cyclic adenosine monophosphate generation in membranes prepared from transplant left ventricles and right ventricles compared with donors. In contrast, guanosine 5'-[beta,gamma-imido]triphosphate, sodium fluoride and forskolin, which stimulate adenylyl cyclase through nonreceptor/G protein sensitive mechanisms, yielded similar degrees of adenylyl cyclase stimulation in the two groups, and both pertussis toxin- and cholera toxin-catalyzed adenosine diphosphate ribosylation were not altered in transplanted left ventricles. CONCLUSIONS: These data indicate that the transplanted human heart exhibits an up regulation of functional beta 2-adrenergic receptors. PMID- 8245348 TI - Aortofemoral transfer function: a method to determine the instantaneous aortic valve gradient in aortic valve stenosis. AB - OBJECTIVES: This study was performed to evaluate the use of synthesized ascending aorta pressure, calculated from femoral artery pressure using an aortofemoral transfer function, in the assessment of aortic valve stenosis. BACKGROUND: Measurement of an accurate aortic valve gradient in patients with aortic stenosis often requires simultaneous recordings of ascending aorta and left ventricular pressures. The use of femoral artery pressure is considered to be a poor substitute for ascending aorta pressure. However, the aortic pressure wave can be calculated from the femoral artery pressure if the aortofemoral transfer function has been determined. METHOD: Femoral artery pressure from the side arm of an introducer sheath and ascending aorta pressure are recorded simultaneously and the data stored in a personal computer. An aortofemoral transfer function is determined from the ratio of the Fourier components of aortic and femoral pressures. Left ventricular and femoral artery pressures are then recorded. Using the previously determined transfer function, the simultaneous ascending aorta pressure is calculated from the femoral pressure. RESULTS: Ascending aorta pressure waveforms estimated from femoral artery pressure closely resembled the simultaneously recorded ascending aorta pressure. Mean aortic valve gradients calculated from the synthesized aortic pressure correlated well with the gradient measured from direct recordings of aortic pressure (r = 0.98). There was also a good relation between valve areas (r = 0.93) and valve resistances (r = 0.98) calculated using the two methods. CONCLUSIONS: Using current computer technology, accurate aortic valve gradients can be rapidly calculated using femoral artery pressure as a substitute for ascending aorta pressure. This technique will reduce the need and risks of multiple catheters to determine aortic valve gradients. PMID- 8245349 TI - Late outcome of unsupported annuloplasty for rheumatic mitral regurgitation. AB - OBJECTIVES: The aim of this study was to evaluate medium- and long-term (range 4 months to 17 years) clinical results in a series of patients treated surgically by unsupported mitral annuloplasty. BACKGROUND: Mitral valve regurgitation has usually been treated by valve replacement or ring annuloplasty. A few series have reported plastic repair procedures without annular support or remodeling. Furthermore, in rheumatic lesions the results have been inferior to those in degenerative mitral insufficiency, and the majority of previous reports have provided information on short- or medium-term follow-up. METHODS: One hundred fifty-four patients were operated on (55 male [36%] and 99 female [64%]). The mean age +/- SD was 36 +/- 16 years (range 5 to 73). Associated lesions comprised 47 aortic and 21 tricuspid valve lesions and 2 atrial septal defects. Patients with concomitant mitral stenosis were not included. Preoperative functional class was I or II in 19% and III or IV in 81%. The cardiothoracic ratio was 0.61 +/- 0.10. All patients underwent an unsupported mitral annuloplasty procedure in which the mural portion of the annulus was reduced by applying two buttressed mattress sutures at the commissures without compromising the width of the septal leaflet. When necessary, additional chordal procedures were performed. No patients received ring or posterior annular support. RESULTS: The early mortality rate was 1.9% (three patients; one of the three died of myocardial failure and two of pulmonary thromboembolism). The late mortality rate was 5.8% (nine patients; three of the nine died of myocardial failure, one each of septicemia, pulmonary thromboembolism and sudden arrhythmic death and three of unknown causes). Twenty-eight patients (18.2%) were reoperated on because of mitral valve dysfunction and 2 (1.3%) because of prosthetic aortic valve dysfunction. A residual late systolic murmur was present in 48% of patients. Late complications were systemic thromboembolism in 5.8% (one third with an aortic valve prosthesis), infective endocarditis in 1.3% and pulmonary thromboembolism in 0.6%. Postoperative functional class was I or II in 84% and III or IV in 16%. Cardiothoracic ratio was 0.58 +/- 0.10. Actuarial probability of late survival was 79.5 +/- 5.3% at 10 years and 71.0 +/- 7.4% at 14 years. Event-free survival was 67.9 +/- 8.9% at 10 years and 56.1 +/- 11.7% at 14 years. CONCLUSIONS: Rheumatic mitral regurgitation can be effectively treated by annuloplasty without prosthetic annular support, with late results comparable to those obtained with more complicated procedures. This observation is particularly important for treatment of children and young adult patients. PMID- 8245350 TI - Natural history of idiopathic dilated cardiomyopathy: effect of referral bias and secular trend. AB - OBJECTIVES: The current study was designed to determine the effect of secular trend and referral bias on the natural history of idiopathic dilated cardiomyopathy. BACKGROUND: In a previous study of 104 patients with idiopathic dilated cardiomyopathy conducted in a referral population at the Mayo Clinic between 1960 and 1973, the 1- and 5-year mortality rates were 31% and 64%, respectively. A recent study of 40 patients with idiopathic dilated cardiomyopathy conducted in a population-based cohort at the Mayo Clinic between 1975 and 1984 reported 1- and 5-year mortality rates of 5% and 20%, respectively. We hypothesized that improvements in diagnosis and therapy have occurred since the original referral cohort was described and that these improvements have altered the apparent natural history of the disease. We refer to this effect as secular trend. Alternatively, the presence of more advanced disease in the referral population (referral bias) may also contribute to the differences in survival. METHODS: Two sequential referral populations with idiopathic dilated cardiomyopathy seen at the Mayo Clinic between 1976 and 81 (n = 85) and 1982 and 1987 (n = 137) were identified. Outcome was compared between these cohorts and the 1960-1973 referral cohort to examine the effect of secular trend. Outcomes were compared with that of the population-based cohort to examine the effect of referral bias. RESULTS: Survival in the 1976-1981 referral cohort did not differ from that in the 1960-1973 referral cohort, suggesting little impact of secular trend during these time periods. Survival in the more recent 1982-1987 referral cohort was significantly better than that in the earlier referral cohorts, suggesting that improvements in diagnosis and treatment in the 1980s altered the natural history of idiopathic dilated cardiomyopathy. Survival in the 1982-1987 referral cohort was still worse than that of the population-based cohort, suggesting an effect of referral bias on studies of the natural history of idiopathic dilated cardiomyopathy. CONCLUSIONS: The current study demonstrates that secular trend and referral bias affect the apparent natural history of idiopathic dilated cardiomyopathy. Survival in referral patients with this disease is significantly better than previously described. PMID- 8245351 TI - Cardiac involvement in Becker muscular dystrophy. AB - OBJECTIVES: The purpose of this study was to assess the incidence of myocardial involvement and the relation of cardiac disease to the molecular defect at the deoxyribonucleic acid (DNA) or protein level in Becker muscular dystrophy. BACKGROUND: Dystrophin gene mutations produce clinical manifestations of disease in the heart and skeletal muscle of patients with Becker muscular dystrophy. METHODS: Thirty-one patients underwent electrocardiographic and echocardiographic examination and 24-h Holter monitoring. The diagnosis was established by neurologic examination, dystrophin immunohistochemical assays or Western blot on muscle biopsy, or both, and DNA analysis. RESULTS: Electrocardiographic and echocardiographic findings were abnormal in 68% and 62% of the patients, respectively. Right ventricular involvement was detected in 52%. Left ventricular impairment was observed either as an isolated phenomenon (10%) or in association with right ventricular dysfunction (29%). Right ventricular disease was manifested in the teenagers, and an impairment of the left ventricle was observed in older patients. Right ventricular end-diastolic volumes were significantly increased compared with those in a control group. The left ventricular ejection fraction was significantly lower in older patients than in control subjects or younger patients. Life-threatening ventricular arrhythmias were detected in four patients. No correlations were found between skeletal muscle disease, cardiac involvement and dystrophin abnormalities. In our patients, exon 49 deletion was invariably associated with cardiac involvement. Exon 48 deletion was associated with cardiac disease in all but two patients. CONCLUSIONS: The cardiac manifestation of Becker muscular dystrophy is characterized by early right ventricular involvement associated or not with left ventricular impairment. Exon 49 deletion is associated with cardiac disease. PMID- 8245352 TI - Differentiation of constrictive pericarditis from restrictive cardiomyopathy by Doppler transesophageal echocardiographic measurements of respiratory variations in pulmonary venous flow. AB - OBJECTIVES: The purpose of this study was to test the utility of measuring respiratory variation in pulmonary venous flow by transesophageal echocardiography. BACKGROUND: Respiratory variation of atrioventricular and central venous flow velocities by Doppler echocardiography has been used to differentiate constrictive pericarditis from restrictive cardiomyopathy. METHODS: We performed pulsed wave Doppler transesophageal echocardiography of the left or right pulmonary veins in 31 patients with diastolic dysfunction. Fourteen patients had constrictive pericarditis, and 17 had restrictive cardiomyopathy. We measured the pulmonary venous peak systolic and diastolic flow velocities and the systolic/diastolic flow ratio with transesophageal echocardiography during expiration and inspiration. The percent change in Doppler flow velocity from expiration to inspiration (%E) was calculated. RESULTS: Pulmonary venous peak systolic flow in both inspiration and expiration was greater in constrictive pericarditis than in restrictive cardiomyopathy. The %E for peak systolic flow tended to be higher in constrictive pericarditis (19% vs. 10%, p = 0.09). In contrast, pulmonary venous peak diastolic flow during inspiration was lower in constrictive pericarditis than in restrictive cardiomyopathy. The %E for peak diastolic flow was larger in constrictive pericarditis (29% vs. 16%, p = 0.008). The pulmonary venous systolic/diastolic flow ratio was greater in constrictive pericarditis in both inspiration and expiration. The combination of pulmonary venous systolic/diastolic flow ratio > or = 0.65 in inspiration and a %E for peak diastolic flow > or = 40% correctly classified 86% of patients with constrictive pericarditis. CONCLUSIONS: The relatively larger pulmonary venous systolic/diastolic flow ratio and greater respiratory variation in pulmonary venous systolic, and especially diastolic, flow velocities by transesophageal echocardiography can be useful signs in distinguishing constrictive pericarditis from restrictive cardiomyopathy. PMID- 8245353 TI - Modified technique for balloon valvuloplasty of critical pulmonary stenosis in the newborn. AB - OBJECTIVES: We report our experience in eight consecutive neonates who underwent attempted balloon dilation as an initial therapy for critical valvular pulmonary stenosis, and we review in detail technical modifications that improved the success rate. BACKGROUND: Balloon dilation of the pulmonary valve has become the treatment of choice for valvular pulmonary stenosis in children and adults. There are few reports of its effectiveness in critical pulmonary stenosis in the newborn. In this setting, application of the technique of balloon dilation has been limited by the ability to advance the necessary guide wires and catheters across the stenotic, often near-atretic, pulmonary valve. METHODS: The pulmonary valve was crossed in all patients. When this could not be accomplished with an end-hole catheter, a soft guide wire was advanced directly across the pulmonary valve through the end-hole catheter positioned in the right ventricular outflow tract below the valve. Initial predilation was achieved in all patients by using a coronary dilation catheter in an effort to facilitate introduction of the definitive balloon dilation catheter. Definitive dilation with a balloon diameter of > or = 110% of the diameter of the pulmonary valve annulus was possible in six patients. RESULTS: Right ventricular pressure declined from a mean value of 108 +/- 32 mm Hg to a mean value of 49 +/- 11 mm Hg after balloon dilation, with no change in heart rate or aortic pressure in these six patients after definitive balloon dilation. CONCLUSIONS: The results of this small series suggest that critical valvular pulmonary stenosis in the newborn can be successfully treated by transluminal balloon valvuloplasty. PMID- 8245354 TI - Influence of ventricular morphology on diastolic filling performance in double inlet ventricle after the Fontan procedure. AB - OBJECTIVES: The purpose of this study was to define the influence of dominant chamber morphology on ventricular performance after the Fontan procedure in patients with double-inlet ventricle. BACKGROUND: Previous studies have reported the impact of ventricular morphology on preoperative ventricular performance and surgical outcome. However, the influence on postoperative ventricular performance has not been addressed. METHODS: Twenty-six clinically asymptomatic patients > 1 year after repair (mean age at procedure 6.1 +/- 3.7 years) were evaluated with ventricular cineangiography and radionuclide blood pool studies (18 with a dominant left ventricular morphology [LV group], 8 with a dominant right ventricular morphology [RV group]) and compared with normal control subjects. RESULTS: Ventricular volume, mass and systolic variables were similar between patient groups. In the LV group, however, the mass/volume ratio was significantly elevated compared with values in control subjects (1.11 +/- 0.28, 0.97 +/- 0.19, p < 0.05), whereas this ratio in the RV group (0.90 +/- 0.11) was within the normal range and significantly lower than that in the LV group (p < 0.05). Mean right atrial and pulmonary artery pressures in the RV group were significantly higher than those in the LV group (p < 0.05). Peak filling rates (2.87 +/- 0.70, 2.41 +/- 1.15 and 3.84 +/- 0.51 end-diastolic volume/s [LV and RV groups and control subjects, respectively]) were significantly lower in both groups than in control subjects (p < 0.001), without intergroup difference. CONCLUSIONS: Ventricular filling abnormalities after atrial to pulmonary anastomosis are common regardless of the type of dominant ventricular morphology, and these abnormalities in patients with dominant right ventricular morphology do not coexist with ventricular hypertrophy. Such diastolic abnormalities may be related to either intrinsic myocardial or acquired factors, not to excessive hypertrophy alone. Those differences may become clinically more apparent with longer follow up and may raise concerns over the long-term course. PMID- 8245355 TI - Echocardiographic predictors of left ventricular outflow tract obstruction after repair of interrupted aortic arch. AB - OBJECTIVES: This study was designed to identify preoperative echocardiographic predictors of left ventricular outflow tract obstruction after repair of interrupted aortic arch and ventricular septal defect closure. BACKGROUND: Left ventricular outflow tract obstruction becomes apparent in nearly 50% of patients after repair of interrupted aortic arch and ventricular septal defect closure but is seldom recognized preoperatively. METHODS: We analyzed the preoperative echocardiograms of all patients with interrupted aortic arch who had postoperative echocardiographic or catheterization data available. Thirty-seven infants (aged 1 day to 10 months, median 5 days) were included. Off-line measurements were performed on hard copies of selected images. The cross sectional area (indexed to body surface area) and diameters (indexed to the square root of body surface area) of the left ventricular outflow tract; ascending and descending aorta; ventricular septal defect; and mitral, aortic and pulmonary valves were compared with outcome by using analysis of variance. Outcome was classified according to development of postoperative left ventricular outflow tract Doppler gradient (Group 1 < or = 20 mm Hg, Group 2 > 20 mm Hg). RESULTS: The cross-sectional area of the left ventricular outflow tract was significantly smaller in patients who did than in those who did not develop subaortic obstruction ([mean +/- SD] 0.64 +/- 0.25 vs. 1.7 +/- 1.01 cm2/m2, p < 0.004). Left ventricular outflow tract and aortic valve diameters and aortic valve area were not predictive of postoperative left ventricular outflow tract obstruction. Incidence of postoperative left ventricular outflow tract obstruction was lower (p < 0.03) in interrupted aortic arch type A (0 of 6) than in type B (15 of 31). The incidence of aberrant right subclavian artery was lower (p < 0.02) in Group 1 (6 of 22) than in Group 2 (10 of 15). CONCLUSIONS: The preoperatively measured cross-sectional area of the left ventricular outflow tract is significantly smaller in patients with interrupted aortic arch who develop subaortic obstruction postoperatively, with a left ventricular outflow tract area < or = 0.7 cm2/m2 being a sensitive predictor. Aortic arch anatomy (i.e., type of interrupted aortic arch and presence of aberrant right subclavian artery) is also predictive of postoperative left ventricular outflow tract obstruction, possibly by influencing the volume of blood flow across the left ventricular outflow tract. These data should enable preoperative identification of infants who may require surgical relief of subaortic stenosis. PMID- 8245356 TI - Positron emission tomographic imaging of cardiac sympathetic innervation using 6 [18F]fluorodopamine: initial findings in humans. AB - OBJECTIVES: This study evaluated the safety, efficacy and validity of 6 [18F]fluorodopamine positron emission tomographic scanning of cardiac sympathetic innervation and function in humans. METHODS: Positron emission tomography (PET) scans, arterial blood and urine were obtained after a 3-min intravenous infusion of 6-[18F]fluorodopamine (1 to 4 mCi, 188 to 809 mCi/mmol) in healthy volunteers, with or without pretreatment with oral desipramine to inhibit neuronal uptake of catecholamines. RESULTS: 6-[18F]Fluorodopamine PET scanning visualized the left ventricular myocardium. Blood pressure increased slightly and transiently. The estimated absorbed radiation dose to the main target organ, the wall of the urinary bladder, was 0.8 to 1.0 rad/mCi of injected 6-[18F]fluorodopamine. By 24 h after the injection, the main 6F-compound in urine was 6F-vanillymandelic acid, a metabolite of 6F-norepinephrine. Desipramine attenuated accumulation of myocardial 6-[18F]fluorodopamine-derived radioactivity and plasma 6F dihydroxyphenylacetic acid. CONCLUSIONS: 6-[18F]Fluorodopamine produces negligible hemodynamic effects and acceptable radiation exposure at doses that visualize the left ventricular myocardium. Sympathetic nerves take up 6 [18F]fluorodopamine, which is translocated from the axoplasm into storage vesicles, where is it beta-hydroxylated to the fluorinated analogue of the sympathetic neurotransmitter norepinephrine. Therefore, the basis for visualization of myocardium after 6-[18F]fluorodopamine injection in humans is radiolabeling by 6-[18F]fluorodopamine and 6-[18F]fluoronorepinephrine of vesicles in sympathetic terminals. 6-[18F]Fluorodopamine PET scanning provides a novel means for assessing sympathetic innervation and function noninvasively in the human heart. PMID- 8245357 TI - Estimation of left ventricular filling pressures using two-dimensional and Doppler echocardiography in adult patients with cardiac disease. Additional value of analyzing left atrial size, left atrial ejection fraction and the difference in duration of pulmonary venous and mitral flow velocity at atrial contraction. AB - OBJECTIVES: The purpose of this study was to determine whether left atrial size and ejection fraction are related to left ventricular filling pressures in patients with coronary artery disease. BACKGROUND: In patients with coronary artery disease, left ventricular filling pressures can be estimated by using Doppler mitral and pulmonary venous flow velocity variables. However, because these flow velocities are age dependent, additional variables that indicate elevated left ventricular filling pressures are needed to increase diagnostic accuracy. METHODS: Echocardiographic left atrial and Doppler mitral and pulmonary venous flow velocity variables were correlated with left ventricular filling pressures in 70 patients undergoing cardiac catheterization. RESULTS: Left atrial size and volumes were larger and left atrial ejection fractions were lower in patients with elevated left ventricular filling pressures. Mean pulmonary wedge pressure was related to mitral E/A wave velocity ratio (r = 0.72), left atrial minimal volume (r = 0.70), left atrial ejection fraction (r = -0.66) and atrial filling fraction (r = -0.66). Left ventricular end-diastolic and A wave pressures were related to the difference in pulmonary venous and mitral A wave duration (both r = 0.77). By stepwise multilinear regression analysis, the ratio of mitral E to A wave velocity was the most important determinant of pulmonary wedge (r = 0.63) and left ventricular pre-A wave (r = 0.75) pressures, whereas the difference in pulmonary venous and mitral A wave duration was the most important variable for both left ventricular A wave (r = 0.75) and left ventricular end diastolic (r = 0.80) pressures. The sensitivity of a left atrial minimal volume > 40 cm3 for identifying a mean pulmonary wedge pressure > 12 mm Hg was 82%, with a specificity of 98%. CONCLUSIONS: Left atrial size, left atrial ejection fraction and the difference between mitral and pulmonary venous flow duration at atrial contraction are independent determinants of left ventricular filling pressures in patients with coronary artery disease. The additive value of left atrial size and Doppler variables in estimating filling pressures and the possibility that left atrial size may be less age dependent than other mitral and pulmonary venous flow velocity variables merit further investigation. PMID- 8245358 TI - Reliability of echocardiographic measurements of myocardial perfusion using commercially produced sonicated serum albumin (Albunex). AB - OBJECTIVES: The purpose of this study was to assess the reproducibility and pitfalls of intracoronary and aortic root sonicated albumin injections, using time-intensity curves, in a large sample of normal dogs. BACKGROUND: The utility of a new myocardial contrast echocardiographic agent, sonicated serum albumin (Albunex), is currently under investigation. However, the reproducibility, injection techniques and general pitfalls of this contrast agent have not been well characterized. METHODS: We administered sequential intracoronary and aortic root injections (518 injections) of sonicated albumin in 25 closed chest normal dogs to measure the effectiveness and reproducibility of this product. Time intensity curves, as a measure of myocardial perfusion, were derived and quantified using an on-line videodensitometric analysis system and two dimensional echocardiography. Measurements included peak intensity, area under the curve, half-time of descent, alpha-parameter and transit time within a 31- x 31-pixel "region of interest" in the anterior septum. Analyses provided 80% power and a type I error protection of 95%. RESULTS: The best reproducibility of the variables was half-time of descent for aortic root injections (coefficient of variation [CV] 20%) and peak intensity for intracoronary injections (CV 25%), whereas aortic root area under the curve showed the most variability (CV 41%). Analysis of variance for repeated measures of serial intracoronary and aortic root injections showed no significant systematic variability within subjects for the measured variables. In a comparison between intracoronary and aortic root injection sites, paired t tests showed no significant difference for mean values between these two techniques. There was also no statistically significant difference between manual versus power intracoronary injections. Finally, there was no significant difference among three injection rates (1, 2 and 3 ml/s) in paired intracoronary injections, nor was there a difference among injection rates in paired aortic root injections, except for a lower peak intensity with a 1-ml/s injection rate compared with a 2-ml/s injection rate (p = 0.01). Potential pitfalls include preparation of sonicated albumin, delivery techniques and measurement variables. CONCLUSIONS: We conclude that the results of serial injections of sonicated albumin show no systemic change or trend in normal dogs. Both intracoronary and aortic root injections at standard injection rates by hand or power injector can be used to quantify time-intensity curves, as measure of myocardial perfusion, with similar variability ranging from 20% to 41%. These results are important in the human model, especially after coronary interventions. PMID- 8245359 TI - Femoral vein delivery of contrast medium enhances transthoracic echocardiographic detection of patent foramen ovale. AB - OBJECTIVES: We postulated that femoral vein delivery of contrast medium because of streaming, might enhance precordial echocardiographic detection of patent foramen ovale. BACKGROUND: Although precordial contrast echocardiography is widely used to diagnose patent foramen ovale, this method is limited by poor sensitivity. Previous investigators have demonstrated enhanced detection of atrial defects by the dye-dilution technique after delivery of contrast medium into the inferior rather than the superior vena cava. METHODS: Transthoracic contrast examinations were performed in a randomly selected group of 70 patients (without previous history of cerebral or systemic embolus) undergoing cardiac catheterization. Paired contrast agent injections (10 ml dextrose in water/0.25 ml air) were administered from an upper extremity vein and femoral vein in each patient during spontaneous respiration, cough and Valsalva maneuvers. Studies were interpreted by an experienced echocardiographer unaware of the sequence and site of injections. Positive studies were semiquantitatively graded from +1 (minimal left ventricular opacification) to +4 (intense left ventricular opacification). Catheterization and echocardiographic assessment of patent foramen ovale were compared in 21 subjects. RESULTS: Patent foramen ovale was detected significantly more often during femoral vein versus upper extremity contrast delivery (23 of 70 patients [prevalence 33%] vs. 9 of 70 patients [prevalence 13%], p < 0.001). The intensity of left ventricular opacification was also greater during femoral vein contrast injection. Precordial echocardiography combined with femoral contrast delivery was significantly more sensitive than cardiac catheterization for assessment of patent foramen ovale (8 of 21 patients vs. 2 of 21 patients, p < 0.05). CONCLUSIONS: Femoral vein contrast delivery significantly enhances the ability of precordial contrast echocardiography to diagnose patent foramen ovale. Physiologic patency of the foramen ovale is more common (prevalence 33%) than previously documented. PMID- 8245360 TI - Re-expansion of balloon-expandable stents after growth. AB - OBJECTIVES: The purpose of this study was to evaluate the feasibility of re expansion of balloon expandable intravascular stents and to examine the gross and histologic effects of re-expansion on vascular integrity. BACKGROUND: Intravascular stents have been used successfully as an adjunct to balloon dilation of congenital pulmonary artery branch stenosis and postoperative stenosis of the pulmonary arteries in children. However, use of rigid stents in children could result in development of relative stenosis at the site of stent implantation with subsequent growth of the child. METHODS: Stainless steel "iliac" stents were placed in the thoracic aorta of 10 normal juvenile swine by a transcatheter technique. Angiography and re-expansion were performed at a mean of 11 weeks (n = 9) and again at 18 weeks (n = 5). After euthanasia, the aortic specimens were removed for gross and histologic examination. RESULTS: Stents were successfully implanted in 10 swine. Re-expansion was successfully performed in each animal at 11 weeks and at 18 weeks. Aortic growth produced a relative constriction of the aorta of 20% +/- 10% (mean +/- SD) at the site of stent implantation at both 11 and 18 weeks. Re-expansion produced a significant increase in mean stent diameter from 10.1 +/- 1 mm to 12.3 +/- 1.2 mm at 11 weeks and from 11.2 +/- 0.7 to 13.5 +/- 1.1 mm at 18 weeks after implantation (p < 0.001). Balloon dilation produced a relative increase in stent diameter of 21% +/ 7% at 11 weeks and 18% +/- 4% at 18 weeks. Stent re-expansion was accompanied by plastic deformation of the neointima without neointimal dissection. Where neointima was thick, there was no evidence of neointimal abrasion, but where neointima was thin, areas of localized neointimal abrasion were observed with focal fibrin and platelet adherence to the stent struts. There was no evidence of medial or adventitial hemorrhage or dissection produced by re-expansion. CONCLUSIONS: Re-expansion of intravascular stents is feasible after growth in juvenile swine without significant injury to neointima, media or adventitia. The results of this study support careful and selective use of intravascular stents as an adjunct to balloon dilation of congenital stenoses in children. PMID- 8245361 TI - Intracoronary air-filled albumin microspheres for myocardial blood flow measurement. AB - OBJECTIVES: The aim of this study was to explore the possibility of quantifying coronary blood flow by myocardial contrast echocardiography with air-filled serum albumin microspheres (Albunex). BACKGROUND: Air-filled albumin microspheres have been proposed as an intravascular tracer for the study of myocardial perfusion by contrast echocardiography. METHODS: In six anesthetized open chest dogs, the left circumflex coronary artery was cannulated and perfused by a roller pump with blood from the femoral artery. Both air-filled albumin microspheres (0.4 ml, 2 x 10(8) spheres/ml) and technetium-99m-labeled albumin were injected as a bolus into the coronary cannula at baseline and after treatment with dipyridamole (0.56 mg/kg body weight intravenously for 4 min). Two-dimensional echographic images of the left ventricular short axis were digitized to generate myocardial time intensity curves; myocardial radioactivity was measured by an external detector to generate radionuclide time-activity curves. RESULTS: After dipyridamole, left circumflex coronary artery blood flow (as measured by both the pump and an electromagnetic flow meter) significantly increased (from 1.06 +/- 0.28 to 3.61 +/- 1.43 ml/min per g of myocardium). Peak intensity and rise time of contrast echo curves were able to differentiate baseline myocardial perfusion from coronary hyperemia but did not show any significant correlation with coronary blood flow. A weak inverse correlation with coronary blood flow was provided by myocardial mean transit time of air-filled albumin microspheres (r = 0.33). Conversely, a close inverse correlation with coronary blood flow was obtained by myocardial mean transit time of technetium-99m-labeled albumin (r = 0.95). Myocardial transit time of air-filled albumin microspheres (1.95 +/- 0.60 s) was also markedly shorter than that of labeled albumin (5.35 +/- 3.43 s, p < 0.001) and the measurements were less reproducible. CONCLUSIONS: In this experimental study, coronary blood flow was not adequately quantified by myocardial contrast echocardiography with intracoronary injection of air-filled albumin microspheres. PMID- 8245362 TI - Characterization of double potentials in a functionally determined reentrant circuit. Multiplexing studies during interruption of atrial flutter in the canine pericarditis model. AB - OBJECTIVES: We tested the hypothesis that double potentials recorded during atrial flutter in a functionally determined reentrant circuit reflect activation of the reentrant wave front around an area of functional conduction block. BACKGROUND: The center of the atrial flutter reentrant circuit in the sterile pericarditis canine model is characterized by double potentials. METHODS: We studied 11 episodes of atrial flutter in eight dogs during interruption of atrial flutter while pacing the atria. A multielectrode mapping system was used to record simultaneously from 190 electrodes on the right atrium (location of reentry). RESULTS: Interruption of atrial flutter occurred when the orthodromic wave front from the pacing impulse blocked in an area of slow conduction in the reentrant circuit. The response of the double potential with interruption of atrial flutter depended on the location of the recording site relative to this area of block. Two types of response were seen. When the double potential was recorded orthodromically distal to this area of block, interruption of atrial flutter was associated with disappearance of the second deflection, and continued pacing after interruption of atrial flutter was not associated with reappearance of the second potential. When the double potential was recorded at a site orthodromically proximal to the area of block, interruption of atrial flutter was not associated with disappearance of the second potential, and when rapid atrial pacing was continued, the double potential remained despite disappearance of the atrial flutter reentrant circuit. CONCLUSIONS: Double potentials represent functional conduction block in the center of the reentrant circuit, with each deflection of the double potential reflecting activation on either side of the area of functional block. The data also demonstrate that double potentials are not limited to a reentrant circuit, as they were recorded on either side of an area of block in the absence of such a circuit. PMID- 8245363 TI - Guidelines for percutaneous transluminal coronary angioplasty. A report of the American College of Cardiology/American Heart Association Task Force on Assessment of Diagnostic and Therapeutic Cardiovascular Procedures (Committee on Percutaneous Transluminal Coronary Angioplasty). PMID- 8245364 TI - Scientific misconduct. PMID- 8245365 TI - President's page: a cardiology summit: the ACC and the subspecialty societies. PMID- 8245366 TI - Bedside markers of coronary reperfusion. PMID- 8245367 TI - Vasovagal reactions after heart transplantation. PMID- 8245368 TI - Interaction of propafenone and mexiletine. PMID- 8245369 TI - Pros and cons of treating anorexia with parenteral nutrition. PMID- 8245370 TI - Nutritional management of diabetes in pregnancy. PMID- 8245371 TI - Why children and parents must play while they eat: an interview with T. Berry Brazelton, MD. Interview by Nancy I. Hahn. PMID- 8245372 TI - Nutrition screening and assessment: current practices and dietitians' leadership roles. AB - OBJECTIVE: To ascertain hospital nutrition screening and assessment practices and the need for additional resources to facilitate these processes. DESIGN: A survey questionnaire was developed to ascertain screening protocols, assessment procedures and tools, relevant training and research needs, and respondent characteristics. SAMPLE: The survey was sent to 988 members of Dietitians in Nutrition Support, a dietetic practice group of The American Dietetic Association; 388 (39.3%) responses were used for analysis. STATISTICAL ANALYSIS: Descriptive statistics were used to calculate response frequency. chi 2 Statistics were used to test associations between nutrition assessment practices and association membership, nutrition support team activity, patient load, time allocations, and type and size of hospital. RESULTS: Most institutions had policies for screening (90.2%); 68.7% of dietitians were involved in screening either alone or with others. Assessment practices within hospitals varied widely: 45.9% used a standard form; 66.8% assessed patients at risk; and 10.6% assessed all patients. Typical assessments included height and weight, diagnosis, weight history, albumin, food intolerance, and allergies. Social histories and sophisticated methods such as indirect calorimetry were used infrequently. Most assessments (54.5%) took 31 to 59 minutes; 35.0% of the dietitians spent more than half their time doing assessments. A large percentage (48.7%) of respondents viewed the dietitian as having greatest influence on assessment policies; others thought physicians (23.2%), administrators (15.7%), nurses (6.2%), or pharmacists (6.5%) had the greatest influence. APPLICATIONS/CONCLUSIONS: Recommended improvements include improved standards of practice, development of written protocols and practice guidelines, better use of human resources, standardization of assessment procedures throughout the profession, and empowerment of dietitians to assume a stronger leadership role in this dimension of dietetics practice. PMID- 8245373 TI - The Dietary Intervention Study in Children (DISC): dietary assessment methods for 8- to 10-year-olds. AB - OBJECTIVES: The dietary assessment methods used in the Dietary Intervention Study in Children (DISC) are described and the rationale, validity, and/or general usefulness of each are discussed. DESIGN: DISC is the first multicenter, randomized, clinical trial to study the feasibility and long-term efficacy, safety, and acceptability of a fat-moderately diet in 8- to 10-year-old prepubescent children with moderately elevated plasma low-density lipoprotein cholesterol (LDL-C) levels. Final data collection for the original study (DISC I) occurred December 1, 1993; continued intervention and follow-up (DISC II) will extend beyond 1997. SETTING: Six clinical centers across the country participate in DISC. SUBJECTS: Preadolescent boys and girls with fasting LDL-C levels between the 80th and 98th age-specific and sex-specific percentiles established by the Lipid Research Clinics were eligible for the study. The feasibility phase included 140 children who were then enveloped into the full-scale trial. Baseline dietary data for 652 randomized children in the full-scale trial and 6-month results for the feasibility cohort are reported. INTERVENTIONS: Dietary assessment involved several elements: (a) determining eligibility based on consumption of more than 30% of energy from total fat, (b) monitoring adherence to and adequacy of the intervention diet, (c) evaluating acceptability of the diet in the intervention group, and (d) determining appropriate foods for the intervention diet. Methods are described for each purpose. MAIN OUTCOME MEASURES: LDL-C differences between the two groups and differences in total and saturated fat intakes as calculated from three 24-hour recalls were the primary outcome measures. Six-month dietary differences in the feasibility group are reported. STATISTICAL METHODS: Baseline group means and 6-month differences in dietary intake are reported for the full-scale trial and feasibility study, respectively. RESULTS: Baseline mean intake from three dietary recalls for the intervention (n = 328) and control (n = 324) groups, respectively, were as follows: energy = 1,759 kcal and 1,728 kcal; total energy from fat = 33.3% and 34.0%; total energy from saturated fat = 12.5% and 12.7%; and total dietary cholesterol = 209 mg and 195 mg. After 6 months of intervention, percentage of energy from total fat and saturated fat was reduced by 5.1% (P = .004) and 2.9% (P < .001), respectively, in this feasibility subset (n = 73) of the intervention group. Essentially no change in these parameters occurred in the control group (n = 67), which demonstrates a measurable difference in reporting between groups. APPLICATIONS/CONCLUSIONS: Results illustrate the feasibility of implementing a variety of dietary assessment methods among preadolescent children without relying primarily on parental reports. PMID- 8245374 TI - Comparison of anthropometric measures of men with HIV: asymptomatic, symptomatic, and AIDS. AB - OBJECTIVE: Anthropometric measures were performed to determine differences in estimated fat mass, lean body mass, and body weight among three groups of men infected with human immunodeficiency virus (HIV). DESIGN: This study was cross sectional. SETTING: Local centers of community services and support groups for persons infected with HIV in the province of Quebec, Canada. SUBJECTS: Thirty seven HIV-positive men were recruited; 11 were asymptomatic (T helper cells [CD4+ count] > 400 cells/mm3), 8 were symptomatic (CD4+ < 400 cells/mm3), and 17 were clinically stable but met the criteria of the Centers for Disease Control and Prevention for acquired immunodeficiency syndrome (AIDS). MAIN OUTCOME MEASURES: Self-reported usual weight, actual weight, body mass index, midarm circumference, and triceps and subscapular skinfolds were recorded. From those we derived the percentage of body fat, the midarm muscle, and fat areas. Daily energy and protein intakes were determined from a 7-day food record. Clinical signs and symptoms were assessed by a structured questionnaire. STATISTICAL ANALYSES PERFORMED: For statistical comparisons, analysis of variance was used, with P < .05 being significant. RESULTS: We found a trend toward a decrease in body weight and in the fat mass indicators as the disease progressed. Lower energy intakes were observed among symptomatic and AIDS groups. The number of nutrition-related clinical signs and symptoms experienced by each individual correlated with the magnitude of weight loss (P < .0004, r = -.69). APPLICATIONS: The findings suggest that anthropometric measures can be used in routine clinical practice to assess changes in body weight and in estimated fat mass among men infected with HIV. Symptoms and energy intakes should be assessed to identify subjects at high risk of greater weight loss. PMID- 8245375 TI - Usual dietary intake and hemoglobin A1 level in patients with insulin-dependent diabetes. AB - OBJECTIVE: To explore epidemiologic evidence for the relationship between dietary intake and glycemic control. DESIGN/SUBJECTS: We examined usual dietary intake, assessed by a food frequency questionnaire, from a random sample (n = 136) of patients who had had insulin-dependent diabetes mellitus for 15 to 21 years. RESULTS: In men, absolute intakes of energy, carbohydrate, protein, fat, and dietary fiber were positively correlated with hemoglobin A1 (HbA1) (P < .05); Spearman correlation coefficients (rs) were .28, .22, .28, .34, and .25, respectively. In women, the correlations were weaker and not significant; rs values were .18, .13, .17, .19, and .16, respectively. When these nutrients were expressed as a percentage of energy or as an amount per 1,000 kcal, only percentage of energy from fat showed a significant association with HbA1--but only in men (rs = .23 for men and .02 for women). Adjustment for body weight, insulin dose, and physical activity using multiple regression analysis did not change the relationship between HbA1 and intakes of energy and fat. Sucrose and alcohol intakes did not show any association with HbA1. CONCLUSIONS: Among men with insulin-dependent diabetes mellitus, the major dietary correlates of poor glycemic control were high intake of energy and percentage of energy from fat. Further investigation is needed to confirm this relationship of energy, fat, and glycemic control. PMID- 8245376 TI - Nutritional status of Chinese-, Korean-, and Japanese-American elderly. AB - OBJECTIVE: To examine the nutritional status of three major subgroups of Asian American elderly. SUBJECTS: The sample consisted of 169 Chinese, 90 Korean, and 50 Japanese elderly who lived in five apartment buildings for senior citizens in Chicago, Ill. MAIN OUTCOME MEASURES: Twenty-four-hour dietary recalls were used to evaluate dietary intake. Anthropometric measures of height, weight, and triceps skinfold thickness were also obtained. RESULTS: Many Asian-American elderly consume an inadequate amount of dietary calcium. A large percentage of Korean elderly also consume inadequate amounts of protein and vitamins A and C. Underweight was more common than obesity among these ethnic elderly groups. APPLICATIONS: The information in this article provides valuable data to the Asian American community for program planning as well as to health providers who work with individual Asian-American elderly to meet their nutrition needs. PMID- 8245377 TI - Nutrition therapy for hepatic glycogen storage diseases. AB - Hepatic glycogen storage diseases (GSD) are a group of rare genetic disorders in which glycogen cannot be metabolized to glucose in the liver because of one of a number of possible enzyme deficiencies along the glycogenolytic pathway. Patients with GSD are usually diagnosed in infancy or early childhood with hypoglycemia, hepatomegaly, poor physical growth, and a deranged biochemical profile. Dietary therapies have been devised to use the available alternative metabolic pathways to compensate for disturbed glycogenolysis in GSD I (glucose-6-phosphatase deficiency), GSD III (debrancher enzyme deficiency), GSD VI (phosphorylase deficiency, which is less common), GSD IX (phosphorylase kinase deficiency), and GSD IV (brancher enzyme deficiency). In GSD I, glucose-6-phosphate cannot be dephosphorylated to free glucose. Managing this condition entails overnight continuous gastric high-carbohydrate feedings; frequent daytime feedings with energy distributed as 65% carbohydrate, 10% to 15% protein, and 25% fat; and supplements of uncooked cornstarch. In GSD III, though glycogenolysis is impeded, gluconeogenesis is enhanced to help maintain endogenous glucose production. In contrast to treatment for GSD I, advocated treatment for GSD III comprises frequent high-protein feedings during the day and a high-protein snack at night; energy is distributed as 45% carbohydrate, 25% protein, and 30% fat. Patients with GSD IV, VI, and IX have benefited from high-protein diets similar to that recommended for patients with GSD III. PMID- 8245378 TI - Validation of 24-hour recalls assisted by food records in third-grade children. The CATCH Collaborative Group. AB - OBJECTIVE: The objective of the study was to validate the use of 24-hour recalls assisted by food records as a dietary assessment tool for use with third-grade children. DESIGN: Trained staff observed children during mealtime at school, and parents observed and recorded what children ate in their presence. The following day children participated in a 24-hour recall interview. Children's ability to recall what they consumed during a 24-hour period was compared with observational data collected during the same period. SETTING: All data were collected in elementary school settings at four sites involved in the Child and Adolescent Trial for Cardiovascular Health. SUBJECTS: The sample of 49 children was self selected, based on parents' willingness to observe and record their child's food intake. MAIN OUTCOME MEASURES: Recalled and observed data for energy and nutrient levels were compared using mean energy and nutrient analysis and quartile classification. In addition, recalled and observed foods were compared by meal type and estimation of portion size. STATISTICAL ANALYSIS PERFORMED: Paired t tests, Pearson and Spearman correlations, and classification analysis were used to compare recalled and observed data. RESULTS: Comparison of observed and recalled food intakes showed no significant differences in percentage of energy from total fat, saturated fat, monounsaturated fat, and polyunsaturated fat or in the amount of sodium consumed, although there were differences in energy intakes. Spearman rank order correlations between recalled and observed nutrients ranged from .45 to .79. A 77.9% agreement was found across all meals in the food items children recalled having consumed compared with those adults actually observed them consuming. CONCLUSIONS: We conclude that the 24-hour recall assisted by food records is a valid method for assessing the dietary intake of children as young as 8 years old for the purpose of group comparison. PMID- 8245379 TI - Hospital infant feeding practices in metropolitan Chicago: an evaluation of five of the 'Ten Steps to Successful Breast-feeding'. PMID- 8245380 TI - Nutrition screening for malnutrition: potential economic impact at a community hospital. PMID- 8245381 TI - Saturated fat intake of elderly women reflects perceived changes in their intake of foods high in saturated fat and complex carbohydrate. PMID- 8245382 TI - Position of The American Dietetic Association: health implications of dietary fiber. PMID- 8245383 TI - 6 x 1 = 1: Part I. PMID- 8245384 TI - Optometry in Russia and the former USSR. PMID- 8245385 TI - Functional ophthalmology in Russia and the former USSR. PMID- 8245386 TI - VOSH-Poland and the optometric dream. PMID- 8245387 TI - An African journey for vision. PMID- 8245388 TI - The growth of professional optometry: focus on optometry in India. PMID- 8245389 TI - International optometry: Let's work together to make change our friend. PMID- 8245390 TI - Onchocerciasis and other eye problems in developing countries: a challenge for optometrists. AB - BACKGROUND: Onchocerciasis, also known as River Blindness, affects about 18 million people around the world, resulting in severe visual impairment or blindness for approximately 2 million. METHODS: The disease is transmitted through the bite of a tiny black fly, which breeds in fast moving rivers and streams. The fly injects immature forms of the parasite worm, Onchocerca volvulus, whose microfilariae migrate to superficial tissues, and may invade any part of the eye. In the eye, living microfilariae may be found in any ocular structure, however, sclerosing keratitis, a severe corneal involvement is the major cause of blindness from the disease. RESULTS: Substantial efforts are currently underway to control the disease in Latin America and equatorial Africa, now that an effective, nontoxic medication, ivermectin, is available. CONCLUSIONS: Optometrists are helping to solve the logistic challenges for treatment of this disease, as most onchocerciasis endemic areas are remote with difficult access. PMID- 8245391 TI - The frequency distribution of ocular disease by age in Imo State Nigeria. AB - BACKGROUND: A total of 600 'first-time served' patients with confirmed ocular pathologies visited our clinic between 1987 and 1988. METHODS: These patients and their ocular diseases were classified according to age, sex, and employment. Gender mix was 53.5 percent male, 46.5 percent female. RESULTS: Of the total disease conditions, 42 percent fell within the 0-30 age group, 44 percent fell within the 31-60 age group and 14 percent fell between 61-90. The relative incidence of ocular disease within this clinic population was: 32.8 percent conjunctivitis; 28.8 percent cataract; 15.5 percent glaucoma; 11.5 percent pterygium; 1.8 percent optic atrophy; and 9.5 percent other. CONCLUSIONS: Caution is made to generalize these data to the population at large since those without financial means were not represented in this study. PMID- 8245392 TI - Constraints to optometric practice in Third World countries. AB - BACKGROUND: Optometry, as a profession in the Third World countries, is in an underdeveloped stage and far from the ideal. Factors responsible for these include literacy level, and the low standard of economic, political, industrial as well as social development. METHODS: Comparisons of various aspects of selected countries bring these factors into clearer focus. RESULTS: Awareness of these problems will lead to possible solutions arising from recognition of need for proper orientation, a focus of interest, manpower, with professional and economic resources in the eye care delivery of the Third World. CONCLUSIONS: The ideal situation can be achieved only when the problems are well defined. PMID- 8245393 TI - Ocular and facial dimensions of male Arab children. AB - BACKGROUND: Photographic measurements of facial and ocular distances of 200 normal male Arab children have been undertaken. METHODS: Measurements include inter inner canthal distance (IICD), inter outer canthal distance (IOCD), length of the palpebral fissure (LPFOD), width of the palpebral fissure (WPFOD), horizontal corneal diameter (HVDOD), vertical brow-nasal distance (VBND), interpupillary distance (IPD), bridge height (BH) and frontal angle (AANG). RESULTS: Percentile values of the measured distances have been given. These values will aid in syndrome diagnosis. CONCLUSIONS: They will also be useful to manufacturers of spectacle frames and lenses. Our values have been compared to those obtained from young males of other racial backgrounds. PMID- 8245394 TI - Exposure keratopathy in a mentally retarded adult population. AB - BACKGROUND: Mentally retarded patients are often found to have exposure corneal disease. No study could be found that documented the prevalence of this corneal disorder among these patients. METHODS: A retrospective study was conducted on a population of 210 mentally retarded adults to determine the prevalence of clinically significant exposure keratopathy. RESULTS: 17.6 percent of this population was found to have this disorder mostly due to incomplete blinking. CONCLUSIONS: Exposure keratopathy was found to be a significant ocular health problem among this population. Since many of these people cannot verbalize their discomfort it is recommended that the optometrist tailor his or her examination so this disorder is not overlooked and untreated. PMID- 8245395 TI - Optometric education and optometry in Africa. AB - BACKGROUND: For the vast majority of people in Africa, eye care services are not available. Optometrists have long been practicing in parts of Africa, but optometric teaching institutes have only recently appeared in several African nations. METHODS: Utilizing interviews, site visits, and a literature review this article presents a survey of the current status of optometric education, the profession of optometry, and eye care needs in Africa. RESULTS AND CONCLUSIONS: Four countries in Africa have optometric teaching institutes: South Africa, Nigeria, Tanzania, and Ghana. Most of these programs are struggling in difficult economic and/or political environments. The difficulties facing these programs are likely to remain, but the recent dramatic developments in communication and cooperation between these four countries will be a positive force towards seriously addressing the need for more optometric practitioners throughout the African continent. PMID- 8245396 TI - Creutzfeldt-Jakob disease: assessment and management. AB - 1. Creutzfeldt-Jakob disease (CJD) is a progressive neurologic disease that occurs in approximately one in one million individuals annually. 2. CJD affects late middle-aged and elderly individuals, with a peak occurrence between the ages of 55 and 75. 3. The disease is thought to be caused by an unknown virus that enters the central nervous system, causing neuron destruction and replacing the destroyed neurons with plaques. 4. There is no known cure or treatment for CJD, and the time span from onset of symptoms until death is 6 to 12 months. PMID- 8245397 TI - Staff attitudes: monitoring technology in long-term care. AB - 1. Overall, care staff members displayed positive attitudes toward resident monitoring technologies, especially staff members working in facilities with more of this technology. 2. It is necessary to place nurses and other front-line health care workers in the center of the policymaking process that determines the technological versus human composition of any long-term care facility. 3. The ethnicity of the health care worker, and to a lesser extent their age and education, are more important than the job status in determining attitudes toward monitoring technology. 4. Comprehensive evaluations of technology used in care facilities should include the overall value of the technology, its impact on a facility's liability, its effect on the quality of care on residents, and whether use of the devices in question is ethical. PMID- 8245398 TI - Nursing knowledge: acute postoperative pain management in the elderly. AB - 1. The inadequate management of acute postoperative pain among adults is well documented. Studies have shown that 75% or more of hospitalized adult patients following surgery suffer moderate or intense pain even with the use of analgesics. Also, physicians under-prescribe narcotic analgesics and nurses administer less than the patient could receive. 2. Nurses' ineffective approaches to the management of pain have been attributed to inappropriate fears of addiction and respiratory depression, rigid attitudes regarding what constitutes adequate pain relief, and misunderstandings about the physiologic and psychologic components of pain. 3. The results of this study support McCaffery's (1989) finding that nurses do not understand the effective use of narcotics in relation to pain management. PMID- 8245399 TI - A vulnerable population: multiproblem older adults in acute care. AB - 1. The older patient has unique needs and vulnerabilities that pose a challenge to nursing. 2. Excess nosocomial infections, incontinence, confusion, activity limitations, skin breakdown, and increased posthospitalization mortality are potential negative outcomes of older adult hospitalization. 3. There is some evidence of age bias in clinical decisions made regarding the older patient. 4. Additional preparation in gerontological nursing, increased sensitization to the needs of older patients, more specialized units, and greater emphasis on basic needs in the acute care setting are necessary to enhance quality nursing care for the frail older patient. PMID- 8245400 TI - Interventions to improve oral care: the nursing assistant's role. PMID- 8245401 TI - Our image. PMID- 8245402 TI - Nursing research: quality of life and perceived health in the elderly. AB - 1. Studies have shown that self-perceived health is significantly related to quality of life for older persons. 2. Terminology regarding quality of life and perceived health was found to be inconsistent in the gerontological nursing research reviewed. 3. Outcomes of the research related to quality of life and perceived health fell into three basic categories: predictors found, relationships found, and no relationships found. 4. More experimental research, more replication of research, more research on older males, more use of nursing theory as a framework for research, and more multidisciplinary gerontological research may contribute useful knowledge to the nursing discipline. PMID- 8245403 TI - Abstracts of the 28th annual meeting of the European Association for the Study of the Liver. Paris, France, 1-4 September 1993. PMID- 8245404 TI - Keratan sulfate and dermatan sulfate proteoglycans associate with type VI collagen in fetal rabbit cornea. AB - Keratan sulfate proteoglycan (KSPG) and dermatan sulfate proteoglycan (DSPG) are associated with collagen fibrils in adult rabbit cornea. Because certain cytochemical data suggested that proteoglycans are associated with type VI collagen in the fetal rabbit cornea, we developed polyclonal antibodies specific to the core proteins of rabbit corneal KSPG (lumican and/or fibromodulin) and DSPG (decorin and/or biglycan) and used the antibodies as immunocytochemical probes to determine proteoglycan ultrastructural location. Immunogold particles were associated with Type VI collagen filaments but not with collagen fibrils in fetal and neonate rabbit cornea. Association of corneal KSPG and DSPG with Type VI collagen was immunocytochemically confirmed with monoclonal antibodies to low sulfated keratan sulfate glycosaminoglycan (GAG) and chondroitin-4-sulfate GAG of DSPG. The monospecificity of the polyclonal and monoclonal antibodies and exclusive binding of antibodies to Type VI collagen filaments, together with previous chemical identification of low-sulfated KSPG and DSPG in developing rabbit cornea, support our contention that corneal proteoglycans in developing rabbit cornea are associated with Type VI collagen. PMID- 8245405 TI - Immunogold labeling of calcium ATPase in sarcoplasmic reticulum of skeletal muscle: use of 1-nm, 5-nm, and 10-nm gold. AB - We evaluated the use of immunogold electron microscopy to study the distribution of calcium ATPase in the sarcoplasmic reticulum membrane of skeletal muscle. We examined (a) 1-nm gold labeling, (b) the effect of gold size on immunolabeling, and (c) the densities of gold particles in areas of maximal labeling in fibers from rat extensor digitorum longus and pig gracilis muscles. The technique allowed unequivocal identification of the calcium ATPase. Gold particles of 1 nm were successfully visualized in unstained or lightly stained sections and the density of labeling was about 20 times greater than with 10-nm gold. The average densities in areas of intense labeling were 2878 +/- 139/microns 2 with 5-nm gold and 4310 +/- 276/microns 2 with 1-nm gold. These numbers are similar to the density of particles in freeze-fracture replicas of sarcoplasmic reticulum. The low density of 10-nm gold suggests that the large gold particles hinder binding of secondary to primary antibodies. The difference between 1- and 5-nm gold is explained by the amounts of gold conjugated to the immunoglobulin. The results suggest that there is a one-to-one relationship between secondary immunoglobulins (1-nm or 5-nm gold conjugates) and oligomeric complexes of calcium ATPase. PMID- 8245406 TI - Differential expression of SPARC and thrombospondin 1 in wound repair: immunolocalization and in situ hybridization. AB - SPARC and thrombospondin 1 (TSP-1) are secreted glycoproteins expressed by similar types of cells in culture and in tissues. To compare these two proteins in vivo, we analyzed the differential expression of SPARC and TSP-1 during wound repair. Full-thickness incision wounds were made in rats and biopsied at 12 hr-14 days. Antibodies against SPARC revealed an increased proportion of immunoreactive fibroblastic cells at the wound edge at 3 days with maximal numbers at 7 days. In situ hybridization for SPARC produced results consistent with those of immunohistochemistry. With combined immunohistochemistry and in situ hybridization, some of the macrophages at the wound edge expressed SPARC mRNA. In contrast, immunoreactivity for TSP-1 was extracellular; expression at the wound edge was noted at 12 hr and was maximal at 1-2 days. TSP-1 mRNA was found in the thrombus, but not at the wound edge. In conclusion, SPARC and TSP-1 have contrasting roles during wound healing. SPARC expression from the middle through late stages of repair was consistent with its previously proposed functions in remodeling; in contrast, the transient expression of TSP-1 early in repair might facilitate the action of other proteins in recruitment and/or proliferation of cells in the healing wound. PMID- 8245407 TI - Expression of human mucin genes in respiratory, digestive, and reproductive tracts ascertained by in situ hybridization. AB - In recent years considerable advances have been made in our knowledge of the peptide moiety of human mucins through cDNA cloning. In many diseases disorders in mucin biosynthesis are observed, which result either from changes in the synthesis of the carbohydrate side chains or from differences in the relative expression of the different apomucins, each of which may affect physical properties of the viscous gel. We describe in situ hybridization studies on healthy human mucosae with five different oligonucleotide probes corresponding to each of the human genes known to date that encode secreted mucins, i.e., MUC 2, 3, 4 (HGM nomenclature) and 5B, 5C (proposed name). These genes present a nucleic tandem repeat organization. The choice of oligonucleotide probes was made to amplify the signal by hybridization of many small probes on the same mRNA molecules. A characteristic pattern of mucin gene expression was observed for each mucosa. PMID- 8245408 TI - Endothelin in normal lung tissue of newborn mammals: immunocytochemical distribution and co-localization with serotonin and calcitonin gene-related peptide. AB - We demonstrated immunoreactivity for endothelins (ET)-1, -2, and -3 and for the precursor, big-ET-1, in the pulmonary diffuse neuroendocrine system (PDNES) of newborn cat, rat, hamster, and mouse. ET-like positive neuroepithelial bodies (NEB) were numerous in the intrapulmonary airways and the alveolar parenchyma. Single neuroendocrine cells (NEC) were less often labeled and mainly localized in the larger bronchi. ET-3-reactive neuronal elements were rarely observed. The intensity and number of immunostained NEB were highest for ET-3, followed in declining order by big-ET-1, ET-1, and ET-2. ET-like possessing NEB displayed interspecies differences. We conclude that ET-3 represents a neuroendocrine form of the ET peptide family. NEB expressing several ET isoforms can be grouped into NEB containing either big-ET-1 and ET-1 or ET-3 only. ET-like immunoreactivity was present in a subpopulation of serotonin (5HT)- and/or calcitonin gene-related peptide (CGRP)-positive NEB. As ET, 5HT, and CGRP have potent pulmonary vaso- and/or bronchomotor effects, our observations suggest that they play a separate or synergistic role in regulatory function of the mammalian PDNES, exerting their influence by paracrine, endocrine, and neurocrine pathways or a combination of these. PMID- 8245409 TI - Post-embedding in situ hybridization for localization of viral nucleic acid in ultra-thin sections. AB - We developed an in situ hybridization technique for localization of plant virus RNA in ultra-thin sections of virus-infected, Lowicryl HM20- or osmium-fixed, Araldite-embedded plant leaf tissues. A digoxigenin-labeled in vitro transcript corresponding to 173 nucleotides at the 3' end of bamboo mosaic virus (BaMV) RNA was used as a riboprobe and the hybrids were detected by incubation with sheep antidigoxigenin antibody followed by gold-labeled rabbit anti-sheep IgG. Pre treatment of ultra-thin sections with an etching agent was the critical step in enhancing hybridization signals on Lowicryl-embedded thin sections. Etching combined with K-metaperiodate treatment increased the accessibility of nucleic acids to riboprobes in osmicated Araldite-embedded sections. BaMV RNA was specifically detected within chloroplasts, mitochondria, and nuclei of infected cells. BaMV virions and BaMV-specific electron-dense crystalline bodies were also labeled. The labeling intensity on Lowicryl-embedded samples, in general, was much higher than that on osmicated Araldite-embedded samples. However, our procedure offers the advantage that it permits labeling of viral nucleic acids in tissues already processed for routine EM and should be applicable for in situ labeling of any cellular nucleic acids. PMID- 8245410 TI - Restricted distribution of integrin beta 6 mRNA in primate epithelial tissues. AB - Integrins are cell adhesion receptors that mediate cell-extracellular matrix and cell-cell interactions. Each integrin consists of two glycoprotein subunits (alpha and beta). We have previously described a novel integrin beta-subunit, beta 6, which is expressed in cultured epithelial cells. beta 6 can associate with alpha v to form the fibronectin-binding heterodimer alpha v beta 6. Here we report the tissue distribution of beta 6 integrin mRNA determined by in situ hybridization of a beta 6 cRNA probe with representative frozen tissue sections from a rhesus monkey tissue bank. We detected beta 6 mRNA exclusively in epithelial cells. However, beta 6 mRNA expression varied greatly among different epithelia. High levels of beta 6 mRNA were found only in two very specialized epithelial cell types: a portion of the kidney tubule epithelium, termed macula densa, and the endometrial epithelium of secretory phase uterus. In the endometrium, beta 6 expression was highest in the differentiated epithelium of functional layer glands, suggesting that beta 6 expression can be regulated in a differentiation-dependent manner. beta 6 expression may also depend on the stage in the estrous cycle, since we found much lower beta 6 mRNA levels in a specimen of proliferative phase endometrium. Epithelium in several other tissues, including salivary gland ducts, gall bladder, and epididymis, contained detectable levels of beta 6 mRNA, albeit much lower than in macula densa and endometrium. In other epithelia, including skin and lung, beta 6 mRNA was undetectable. Taken together, these results suggest that in normal adult primates beta 6 expression is regulated in a cell type-specific manner, restricted to a few epithelial tissues. PMID- 8245411 TI - Histochemical localization of copper in the intestine and kidney of macular mice: light and electron microscopic study. AB - Menkes' disease is an X-linked recessive disorder characterized by accumulation of copper in various organs and cells, such as the intestine, kidney, and cultured fibroblasts. Light and electron microscopic localization of Cu was investigated in the intestine and kidney of macular mice, an animal model of Menkes' disease, by a modified sulfide-silver method. Cu was accumulated in the cytoplasm of the absorptive epithelial cells, the vascular endothelium, and secretory granules of the Paneth cells. In kidney the distal tubule cells and glomeruli of both macular and control mice stained faintly, whereas the organelle free cytoplasm in the proximal tubule cells of macular mice stained more intensely than those of controls. The nuclei, mitochondria, and lysosomes of the cells of macular mice hardly stained at all. These findings indicate that Cu is concentrated in the organelle-free cytoplasm of the affected cells of macular mice. This suggests that the Menkes' mutation affects Cu transport from the cytosol to the organelles in the cell. PMID- 8245412 TI - Superiority of in situ hybridization over immunolabeling for detecting DNA on Lowicryl sections: a study on adenovirus-infected cells. AB - We investigated the intranuclear distribution of viral genomes in adenovirus Type 5-infected HeLa cells on ultra-thin sections of Lowicryl K4M-embedded material by immunolabeling of DNA and in situ hybridization with a viral DNA probe. Monoclonal AC-30-10, raised against double- and single-stranded DNA (dsDNA, ssDNA), and HB2 antibodies, raised against only the dsDNA, were used. Both antibodies intensely labeled the condensed host chromatin and the virus-induced substructures containing inactive encapsidated and non-encapsidated viral genomes. The antibodies labeled only slightly those substructures containing replicating and transcribing viral genomes. Viral ssDNA, which is accumulated within well-delineated compact fibrillar structures, was not revealed by AC-30-10 even after elimination of the proteins of the section. In situ hybridization detected all of these types of viral DNA, depending on the protocol used. Therefore, the sensitivity of in situ hybridization for detecting DNA appears superior to immunolabeling with specific antibodies. PMID- 8245413 TI - Distribution of calcitonin gene-related peptide in vertebrate neuromuscular junctions: relationship to the acetylcholine receptor. AB - Calcitonin gene-related peptide (CGRP), regarded by several authors to be involved in maintenance of the acetylcholine receptor, is present in the motor axons of various striated rat muscles. It is present, however, only in motor endplates of several selected striated muscles, where it is located in presynaptic axon terminals of neuromuscular junctions. No immunoreactivity could be seen within synaptic vesicles themselves. In the non-human primate Macaca fasciculata, neuromuscular junctions, including those in the diaphragm, display an intense CGRP reaction. The structure of the simian motor endplates is more elaborate than that of the rat. Amphibian motor nerve endings, both in tetanic and tonic muscles, display CGRP immunoreactivity. In tetanic muscles the CGRP reaction outlines "terminaisons en placque" (true motor end plates) and weakly reacting "terminaisons en grappe" (grape-like endings) in tonic muscles. On supramaximal stimulation of the motor nerve, CGRP is depleted from the affected neuromuscular junctions. Wallerian degeneration of the motor axon results in complete disappearance of CGRP. In most rat muscles in which motor endplates do not normally exhibit CGRP immunoreactivity, e.g., the diaphragm and buccinator muscles, the pre-terminal motor axons are CGRP-positive. After immobilization of such muscles by local bupivacaine injection to rats under brief chloral hydrate anesthesia, CGRP immunoreactivity of the neuromuscular junctions can be elicited because blockade of neuromuscular transmission results in accumulation of CGRP in the endplates. Even more striking is the appearance of CGRP immunoreactivity in normally non-reactive motor endplates during axon regeneration after an experimentally induced Wallerian degeneration of the motor axons. We conclude that CGRP is a regular, genotypically determined component of neuromuscular junctions, present either in a manifest or in a latent form. The latter can be elicited by various experimental approaches. The presence of CGRP in the motor endplate supports the theory that this peptide is instrumental in maintenance of the acetylcholine receptor. PMID- 8245414 TI - A specific fluorogenic reaction for tryptophan residues using isatin in organic solvents. AB - The development of new fluorogenic reactions for specific chemical groups is of increasing interest in cytochemistry. We describe the application of the nonfluorescent compound isatin on methanol-fixed blood smears. When treated with 0.1% isatin in absolute ethanol or acetone for 30 min and observed under violet blue exciting light, eosinophil leucocyte granules show a bright green fluorescence. This fluorogenic reaction is abolished after blocking of tryptophan residues by performic acid oxidation. Spectrofluorometric studies with amino acids in vitro reveal that isatin forms a unique and specific fluorescent product with tryptophan. PMID- 8245415 TI - Polysialosyl glycoconjugates defined by monoclonal antibody M6704 are expressed in human gliomas and embryonic neurons. AB - The distribution of polysialosyl glycoconjugates defined by a monoclonal antibody M6704 (M6704 antigen) and of its O-acetylated counterpart (O-Ac-M6704 antigen) was investigated in human gliomas and in normal adult and fetal central nervous system. Among 32 gliomas, M6704 antigen was confined to the glioma cell surface and cytoplasmic processes in 21 cases, whereas O-Ac-M6704 antigen was expressed in the Golgi region in 22 cases and at the cell surface in seven cases. In adult tissues M6704 antigen was barely detectable, whereas in fetal tissues it was spatially and temporally expressed at the cell surface and along processes of post-mitotic neurons. The O-Ac-M6704 antigen first appeared in the Golgi regions of neurons at 66 gestational days and increased gradually with further maturation to adult levels. Several polysialoglycoproteins, the most dominant being 51 KD, and C-series polysialogangliosides were isolated from gliomas. These results indicate that M6704 antigen is a distinctive component expressed selectively by embryonic neurons during development and by glioma cells. PMID- 8245416 TI - Use of fluorescent latex microspheres (FLMs) to follow the fate of transplanted myoblasts. AB - A potential treatment for Duchenne muscular dystrophy (DMD) is injection of normal myoblasts into dystrophic muscles to induce formation of muscle fibers. To develop this therapy it is important to identify the injected myoblasts and the muscle fibers that they form in the host muscles. Fluorescent latex microspheres (FLMs) were used for this purpose in this study. Normal myoblasts were labeled with FLMs and injected into dystrophin-deficient (mdx) mice. The FLMs clearly indicated the location of injected myoblasts in the host muscle. Muscle fibers containing dystrophin were localized by immunofluorescence and immunoperoxidase. They were observed in clusters near the myoblasts labeled with FLMs. FLMs were also observed in some of these dystrophin-positive fibers in each cryostat section. These results indicate that: labeling myoblasts with FLMs can be used to trace the injected myoblasts in the muscle and to identify the muscle fibers that they formed; injected myoblasts remain near the injected site and do not migrate very far; most of the dystrophin-positive muscle fibers around the injected myoblasts result from fusion of the injected myoblasts; and the low percentage of dystrophin-positive muscle fibers is likely related to limited diffusion and lack of fusion of many injected myoblasts. PMID- 8245417 TI - The mouse chimera during intrauterine stages: immunohistochemical analysis with the C3H strain-specific antibody. AB - Our objective was to establish an immunohistological method for analysis of chimerism in mouse chimeras at embryonic stages with an anti-C3H strain-specific antigen (CSA) antibody. We developed an effective new method to retain CSA antigenicity with good morphology of embryonic tissues by using microwave irradiation (MWI) for pre-fixation, 95% ethanol/1% acetic acid as post-fixative solution, and polyester wax as embedding material. We used a biotinylated mouse monoclonal anti-CSA antibody, peroxidase-avidin, and silver amplification. These procedures were successful in demonstrating the chimerisms in various tissues of C3H<-->Balb/c chimeras at different embryonic stages and postnatal days. In chimeras at Days 7 and 7.5 post coitum (p.c.), both genotypes were clearly identified and well intermingling in every embryonic tissue (embryonic ectoderm, mesoderm, extra-embryonic ectoderm, ectoplacental cone, amnion, and chorion). Chimerisms at Day 14.5 p.c. were also clearly observed in mesencephalon, neural retina, spinal cord, lung, kidney, and liver. We concluded that the present immunohistological procedures for analysis of chimerism during embryonic periods will give us insightful information about dynamic histological changes such as cell proliferation, migration, selection, and death during organogenesis. PMID- 8245418 TI - Workshop on in situ hybridization: what you need to know to get it to work. PMID- 8245419 TI - Fundamental principles of in situ hybridization. AB - In situ hybridization provides invaluable information regarding the localization of gene expression in heterogeneous tissues. The technique is extremely sensitive and can detect the amount of mRNA contained in a single cell. This review provides a starting point for those who wish to begin using in situ hybridization in their own laboratories. The procedure outlined here is based on 35S-labeled riboprobes and has been used with many probes and tissues with a greater than 90% success rate on the first hybridization. The importance of appropriate controls is stressed. Clusters of silver grains after hybridization do not necessarily indicate specific mRNA localization. Regions of the tissue rich in nuclei often appear to cause spurious binding of probes and have high backgrounds often mistaken as positive signals. The most difficult aspect of in situ hybridization is not to get clusters of silver grains on the slide but rather to do the appropriate controlled experiments to ensure that the signal is real and is not due to some artifactual binding of the probe to the tissue. PMID- 8245420 TI - Selection of oligonucleotide probes for detection of mRNA isoforms. AB - In this article we discuss strategies for selecting oligonucleotides to target isoform-specific mRNAs, drawing on our experience with isotopically labeled oligonucleotides for ISH of Na,K-ATPase mRNA alpha- and beta-subunit isoforms. Oligonucleotide probes based on one of these isoforms have a high probability of forming nonspecific hybrids with related isoform mRNAs. The design and selection of isoform-specific ISH and how their nucleotide structure influences hybridization are reviewed, as well as basic principles in identifying and evaluating candidate probes. Controls such as Tm analysis and GC content are evaluated. For distinguishing among multiple isoforms of gene families, choose lowest possible homology between isoforms consistent with other factors that influence probe performance. PMID- 8245421 TI - A method for simultaneous detection of multiple mRNAs using digoxigenin and radioisotopic cRNA probes. AB - We present a simple, reliable method for simultaneous detection of two distinct mRNAs within the same tissue sample by double in situ hybridization histochemistry. Sections are hybridized with a cocktail of radiolabeled and digoxigenin-labeled cRNA probes. The digoxigenin-labeled probe is detected with an alkaline phosphatase (AP)-dependent chromogen reaction and then the radiolabeled probe is detected by conventional autoradiography. The sensitivities of the two detection methods are comparable and demonstrate the feasibility of using double in situ hybridization histochemistry to investigate the co localization and co-regulation of mRNAs that are expressed at relatively low levels. PMID- 8245422 TI - Biotinylated probes for in situ hybridization histochemistry: use for mRNA detection. AB - Biotinylated probes for in situ hybridization (ISH) are now widely used to detect RNAs and viral genomes at the light and electron microscopic levels. Many protocols for ISH with biotinylated probes are now available. The most critical parameters that influence sensitivity and specificity of the procedure are the nature of the probe, the mode of fixation or incorporation of biotin, and the mode of detection of biotin. Biotin can be detected with antibodies or with avidin (or streptavidin) to which is coupled a marker that can be identified under the microscope. Here we review the protocols, pitfalls, advantages, and disadvantages of biotinylated probes for ISH. PMID- 8245423 TI - Fluorescence in situ hybridization using digital imaging microscopy. PMID- 8245424 TI - Detection of messenger RNA and low-abundance heteronuclear RNA with single stranded DNA probes produced by amplified primer extension labeling. AB - We describe a procedure for detection of low-abundance cellular RNAs by in situ hybridization histochemistry, using single-stranded DNA probes produced by amplified primer extension labeling with Taq polymerase. We have used this approach to detect a number of high- and low-abundance RNA species and have found it to be a simple and reproducible method of obtaining sensitive probes for in situ hybridization studies. For example, DNA probes generated by amplified primer extension labeling can detect low-abundance heteronuclear RNAs in individual neurons. Since this procedure does not involve recombinant DNA technology or microbiological facilities, it should prove useful to a wide variety of investigators studying the regulation of gene expression at the cellular level. PMID- 8245425 TI - Fundamentals of quantitative autoradiography by computer densitometry for in situ hybridization, with emphasis on 33P. AB - The goal of quantitative autoradiography (QAR) in in situ hybridization (ISH) is to determine the amount of radioactive oligonucleotide or riboprobe present in the corresponding area of the tissue slice that produced an autoradiographic image. This article discusses (a) some of the considerations related to selection and use of computer image analysis systems for accomplishing this objective, (b) development of 14C plastic autoradiographic standards for ISH QAR with 33P, (c) using QAR to develop Tm curves for ISH probes, and (d) measurement of resolution with video imaging systems for QAR. PMID- 8245426 TI - Estimation of tissue mRNAs by in situ hybridization. AB - We describe a new stereological method for analyzing data derived from the in situ hybridization procedure. This method should prove important, since data summarization in terms of grains per anatomic area by sampling of tissue sections may lead to faulty interpretations. Using computer simulation of measurements taken from a two- and a three-dimensional perspective, we show how the detection of molecular changes can be influenced by multiple structural events. Cell volume, the volume of the structure, and the number of labeled cells in an anatomic structure are all important parameters that must be assessed to obtain accurate results. We then outline in detail a simple and efficient method for estimation of the number of mRNA molecules in cells and in the total structure. By estimation of the abundance of mRNA molecules in three dimensions, we can make better inferences concerning gene expression in a structural context. PMID- 8245427 TI - Nile red labeling of single living cells for contour delineation to quantify and evaluate the distribution of rhodamine 123 with fluorescence image cytometry. AB - Simultaneous study of intracellular quantification and distribution of fluorescent probes is difficult when cell staining is not homogeneous. This occurs after mitochondrial staining with rhodamine 123 (R123). Classical techniques for evaluation of intracellular R123 fluorescence, such as flow cytometry, are based on measurement of the global fluorescence intensity but do not take into account parameters that reflecting cellular distribution of the probe. For simultaneously studying intracellular quantification and distribution of R123 with fluorescence image analysis, we delineated a mask of the cell, generated from a fluorescent image of the plasma membrane stained by nile red (NR). After a preliminary study of the fluorescence characteristics of R123 and NR to avoid artifacts and optimize conditions of staining, quantification and distribution of intracellular R123 studies were performed by superimposition of the mask on the R123 fluorescence image. This protocol was applied to leukemic cells and allowed estimation of individual cell parameters such as mean fluorescence intensity and standard deviation, the latter providing information of the cellular distribution of R123. Moreover, it permitted demonstration of the redistribution of R123 in the whole cell when coincubated in the presence of nigericin. PMID- 8245428 TI - Quantitative electron probe microanalysis of acetylcholinesterase activity in rat brain sections. AB - In this study we determined acetylcholinesterase (AChE) activity by electron probe microanalysis (EPMA). AChE was localized in rat brain sections by a histochemical reaction that produced a Cu-ferricyanide precipitate. This reaction is based on the hydrolysis of acetylcholine substrate with AChE. The copper (iron) concentration in the deposit is proportional to acetylcholine involved in the hydrolysis reaction. Absolute determination of the size of the reaction was calculated from EPMA measurements compared with those made from a dextran standard containing a known amount of Cu(Fe). Electron probe microanalysis allows AChE activity to be assayed quantitatively on thick brain sections with a spatial resolution of 10 microns. PMID- 8245429 TI - Secretory granules and granins in hyperstimulated male rat gonadotropes. AB - Granins are acidic proteins co-localized with peptides in secretory granules of many endocrine cells. They are thought to participate in certain steps of the regulated secretory pathway. This is of particular interest in rat pituitary gonadotropes, which in most cases contain both gonadotropins (follicle stimulating hormone, FSH and luteinizing hormone, LH) and two granins (chromogranin A, CgA and secretogranin II, Sg II). Therefore, we investigated male rat gonadotropes ultrastructurally and for the cellular and subcellular localization of gonadotropins/granins under normal conditions and after stimulation by luteinizing hormone-releasing hormone (LHRH) or castration. Typical gonadotropes of controls contained differently composed secretory granules: small granules showed immunoreactivity for LH and SgII and larger granules were immunoreactive for FSH and CgA and to a lesser extent, for LH. Stimulated gonadotropes showed hypertrophy or hyperplasia and RIA for plasma LH levels showed a 30-70-fold increase. In stimulated cells a third type of secretory granule became prominent. These "intermediate" granules had an electron dense core immunoreactive for LH and SgII, whereas CgA labeling was confined to a less electron-dense outer region. (In stimulated gonadotropes, FSH immunoreactivity could be shown effectively only at the light microscopic level.) Intermediate granules developed from structures resembling condensing vacuoles. They began to exhibit their typical double structure as they budded off from the trans-Golgi network. It therefore appears that CgA and SgII participate in establishing two different routes of the regulated pathway in gonadotropes. Therefore, immunocytochemistry of the granins seems to be a suitable approach to investigating secretory pathways in these endocrine cells. PMID- 8245430 TI - Cellular localization of endothelial alkaline phosphatase reaction product and enzyme protein in the myocardium. AB - Myocardial capillary endothelial cells, arteriolar endothelial cells, and the arterial adventitia show positive alkaline phosphatase (AP) enzyme reaction and immunoreactivity in both rat and human hearts. In guinea pigs, however, capillary endothelial staining is discontinuous and arterial adventitia is negative. The ultrastructural correlate of discontinuous capillary staining is a pronounced labeling of pericytes in guinea pig heart and relatively weak endothelial staining. In rat and human heart, enzyme reaction products are localized mainly on plasma membranes and cytotic vesicles of endothelial cells. Comparison of two strains of rat reveals a more dense deposition of enzyme reaction product along the luminal and particularly along the abluminal plasma membrane of Sprague Dawley rats than of Wistar rats. Quantitative analysis of immunogold labeled anti AP antibody density confirms the pronounced polarity of capillary endothelial cell labeling in Sprague-Dawley rats. More than 80% of total endothelial AP protein in Sprague-Dawley rats is localized over the abluminal plasma membrane and basal lamina, as compared with less than 30% in Wistar rats. Moreover, the total endothelial cell labeling is almost sixfold higher in Sprague-Dawley than in Wistar rats. Total endothelial labeling and proportion of labeling on the abluminal endothelial plasma membrane in human hearts is intermediate between the two strains of rat. The strain and species differences in enzyme distribution could provide important information concerning enzyme function. PMID- 8245431 TI - Comparison of anti-fading agents used in fluorescence microscopy: image analysis and laser confocal microscopy study. AB - To ascertain the ability of commercial and home-made anti-fading media to reduce the decrease of fluorescein isothiocyanate (FITC) fluorescence, we studied the bleaching characteristics of FITC-stained Reh 6 cells mounted in buffered glycerol and in anti-fading media. We measured the intensity of fluorescence over time with a confocal laser scanning microscope and a standard epifluorescence microscope coupled to an image analysis system. Most of the anti-fading media effectively retard fading but each has drawbacks. Better results were obtained with media containing p-phenylenediamine (solutions in buffered glycerol, Vectashield, Fluorstop). However, Mowiol, Slowfade, n-propyl gallate (20 g/liter) were also effective in retarding fading. Most of them, except Mowiol, reduced fluorescence intensity. We concluded that the choice of anti-fading medium would depend on the desired results: a slower decay of fluorescence despite an initial quenching of fluorescence or a lower retardant effect with no decrease in initial fluorescence intensity. Moreover, the combination of Mowiol with another anti fading medium may be a useful compromise when a strong retardant effect is required without marked quenching of the initial fluorescence. PMID- 8245432 TI - Significance of high glucose-6-phosphatase activity in rat oviduct epithelium. AB - To study the origin of glucose in the oviduct fluid, we cytochemically examined glucose-6-phosphatase (G6Pase) activity in rat oviduct. The activity in the whole oviduct was also assayed biochemically. During proestrous, estrous, and metestrous phases, staining reaction for the activity was moderate in the epithelium of the caudal isthmus (CaI) and uterotubal junction (UJ), whereas it was weak in that of the ampulla (A) and cephalic isthmus (CeI). In the diestrous phase, staining reaction in the epithelium of CaI and UJ became strong although it remained weak in that of A and CeI. Reaction product for the activity was localized in the endoplasmic reticulum and nuclear envelope of all cell types in the epithelium. The amount of reaction product in secretory cells was small to moderate in CaI and UJ, and small in A and CeI during proestrus, estrus, and metestrus. In diestrous the amount became abundant in CaI and UJ and moderate in A and CeI. However, the amount in ciliated cells remained small in the four segments during the four phases. The biochemical activity in diestrous was greater than that in proestrus, estrus, or metestrus. This shows that the activity is high in secretory cells in the epithelium of CaI and UJ in the diestrous phase and suggests that the role of the high activity is to release glucose into the oviduct fluid for use by the embryo passing down the CaI and UJ to the uterus. PMID- 8245433 TI - Hepatic lobular patterns of phosphoenolpyruvate carboxykinase, glycogen synthase, and glycogen phosphorylase in fasted and fed rats. AB - The goal of this study was to localize phosphoenolpyruvate carboxykinase (PEPCK), glycogen synthase (GS), and glycogen phosphorylase (GP) in the liver lobule by immunocytochemical techniques and to describe the effects of feeding and fasting on the distribution and quantity of these enzymes. Livers from ad lib fed and overnight fasted normal adult male rats were frozen in liquid nitrogen after transcardial perfusion with 30% sucrose. Serial cryostat sections of tissue were collected on slides, fixed by immersion in 4% paraformaldehyde, and incubated with antibodies against PEPCK, GS, and GP. Antibodies to these enzymes were visualized with a gold-conjugated secondary antibody and a silver enhancement technique. Fed animals demonstrated a periportal to pericentral gradient of PEPCK. Fasting increased the periportal content of PEPCK, induced the midlobular and centrilobular cells to express the enzyme, and steepened the periportal to pericentral gradient. The increase of PEPCK was confirmed by Western blot analysis. GS and GP were distributed throughout the lobule in the fed animal but often showed a centrilobular pattern, and fasting did not alter the lobular distribution of either enzyme. Western blot analysis revealed no changes in the amount of these enzymes in the fed or fasted state. The cellular distribution of the three enzymes is similar to that of hepatic glycogen, in that the immunoreactive material has a clumped appearance in the periportal hepatocytes and is more dispersed in the pericentral cells. On fasting the periportal hepatocytes lose the dense compact localization of the enzymes and the protein becomes more homogeneously distributed throughout the cytosol. Further studies are needed to elucidate the functional significance of the regional heterogeneity of the glycogen-metabolizing enzymes and the molecular mechanisms regulating their gene expression. PMID- 8245434 TI - Expression of the cysteine proteinase inhibitor cystatin C gene in rat heart: use of digoxigenin-labeled probes generated by polymerase chain reaction directly for in situ and northern blot hybridizations. AB - Cystatins represent a widely distributed superfamily of cysteine proteinase inhibitory proteins. We investigated the expression of the cystatin C gene, belonging to the family 2 of cystatins, in the hearts of female rats. Using a highly sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) we have detected cystatin C mRNA in the ventricule and atrium, as well as in liver and submandibular gland. A digoxigenin-labeled cystatin C probe, generated by PCR, hybridized to a single mRNA species of about 700 nucleotides on Northern blots. Northern blot hybridizations established that neither an acute inflammation produced by injection of turpentine nor administration of the beta-adrenergic agonist isoproterenol had an effect on the level of cystatin C mRNA in the heart. In situ hybridizations with digoxigenin-labeled probe localized the expression of the cystatin C gene to cardiac muscle fibers but not to other cardiac cellular elements. Cystatin C may be released by cardiac muscle fibers under physiological and pathological conditions and may modify inflammatory and necrobiotic processes. PMID- 8245435 TI - Controls for immunogold labeling. PMID- 8245436 TI - Intrathymic lymphoid precursor cells during fetal thymus development. AB - Our previous studies have demonstrated the presence, in the adult mouse thymus, of a population of early precursor cells able to give rise to T and B lymphocytes but not myeloid cells. This population of cells expresses low levels of CD4 and has been termed the "low CD4 precursors." All these precursors were found to be c kit positive, and they precede the better known CD4-CD8- precursor stage. In this study, embryonic and neonatal thymuses were examined to see whether a similar low CD4 precursor was part of the pathway of T cell development during ontogeny. A population with the phenotypic characteristics of the adult low CD4 precursor was found from day 15 of embryonic development, although the expression of low levels of CD4 was apparent only from embryonic day 17. Functional tests of these putative precursors showed they had no thymus-reconstituting ability when isolated from thymuses at any time during embryonic life, and very low reconstituting ability even 24 days after birth. These results raise questions about the adult low CD4 precursor as an obligatory stage in the development of T cells in the thymus. PMID- 8245437 TI - Functional effects of CD30 on a large granular lymphoma cell line, YT. Inhibition of cytotoxicity, regulation of CD28 and IL-2R, and induction of homotypic aggregation. AB - Studies are described revealing novel regulatory functions for the lymphocyte activation Ag CD30. A new mAb, C10, reactive with YT cells binds to CD30 and induces inhibition of the cytotoxicity of YT for Raji cells. C10 inhibition of cytotoxicity requires several hours preincubation of YT with C10; the antibody has no effect if added directly to YT cytotoxicity assays. CD30 stimulation by C10 down-regulates CD28 expression on YT by > 80% within 48 h. Because CD28 is required for YT cytotoxicity toward Raji cells and other B7/BB1 bearing targets, it is suggested that inhibition of cytotoxicity of YT is mediated by control of CD28 expression and/or signaling via CD30. Accordingly, conjugation of YT with Raji is only slightly affected by CD30-mediated down-regulation of CD28, and perforin mRNA steady state levels are not changed at all. C10 treatment of YT cells additionally down-regulates the expression CD45 and up-regulates IL-2R p55. Moreover, CD30 stimulation by C10 causes homotypic aggregation of YT. Homotypic aggregation is slow, requiring gene transcription, translation, metabolic energy at elevated temperature (37 degrees C), magnesium ions, and an intact cytoskeleton. These studies offer insights into the function of CD30 as a complex regulator of T cells. PMID- 8245438 TI - Production of soluble IL-4 receptors by murine spleen cells is regulated by T cell activation and IL-4. AB - Many cytokine receptors exist naturally as both membrane-bound and soluble forms. Whereas the membrane receptors have an obvious role in signal transduction, the putative immunoregulatory role played by the soluble receptors remains unclear. Although natural forms of soluble IL-4R (sIL-4R) are known to be present in the biologic fluids of normal mice, the mechanisms regulating the production of sIL 4R have not been characterized. In this study, we have developed an ELISA that allows the measurement of sIL-4R without interference from endogenous IL-4, and have analyzed the effect of cellular activation and several cytokines on the secretion of sIL-4R by murine splenic cells. Although normal spleen cells in culture produced low, but detectable levels of sIL-4R under basal conditions, stimulation with the T cell-mitogens, Con A or soluble anti-CD3 antibodies, caused a 10- to 40-fold increase in the production of sIL-4R. Stimulation of B lymphocytes with LPS, however, did not result in significant up-regulation of sIL 4R secretion. Moreover, IL-4, but not other cytokines, was also a potent inducer of sIL-4R production by spleen cells, even in the absence of other stimuli. Blocking experiments with an anti-IL-4 antibody, 11B11, demonstrated that the effect of T cell-mitogens is partially mediated by endogenously produced IL-4. Cell depletion experiments suggested that although the effect of T cell-mitogens was dependent on the presence of viable T cells, all major cell types including T cells, B cells, and macrophages, either resting or activated, were able to up regulate their secretion of sIL-4R in response to IL-4. Unlike many activities of IL-4, the secretion of sIL-4R by IL-4-stimulated splenic cells was not antagonized by IFN-gamma. These results suggest that the production of sIL-4R is regulated by stimuli leading to T cell activation and IL-4 secretion and are consistent with sIL-4R having an important role in the regulation of IL-4 activity in vivo. PMID- 8245439 TI - Reversion of the SCID phenotype by human T cell grafts. Development of cross species immunocompetence. AB - Due to defective recombinase function, mice with severe combined immunodeficiency (SCID) lack functional lymphocytes and can accept human lymphoid xenografts. Xenografted animals (SCIDhum) are thought to provide a neutral environment for in vivo studies of normal, malignant or HIV-infected human cells. SCIDhum often develop endogenous, EBV+ lymphomas in the graft and in the our study two-thirds of 142 SCIDhum mice did so. Surprisingly, one-third of animals developed reversion of the SCID phenotype rapidly after human T cell engraftment. 90% of tumors occurred in nonrevertant and only 10% in revertant mice. These revertant animals showed immunologic tolerance for normal human B lymphocytes, maintained stable levels of mouse and human IgM and IgG. In addition, they generated competent mouse T cells able to kill transformed (EBV+) but not fresh B cells from the same donor nor unrelated human B cell lines. The tolerance for human lymphoid cells and the cross-species antitumor competence of host T lymphocytes imply unexpected recognition and selection events. Rather than a neutral "bioreactor," these observations mark the SCID host as potentially active participant in a composite immune system generated by xenografting. PMID- 8245440 TI - Influence of membrane Ig receptor density and affinity on B cell signaling by antigen. Implications for affinity maturation. AB - We have initiated a series of experiments to explore the effect of changes in density of the surface Ig receptor (mIg) on Ag:mIg interactions. We transfected into the surface Ig-negative B cell line M12.4 H and L chain constructs known to effect a 10-fold change in antibody binding affinity for the naturally occurring hapten phosphocholine (PC). Two sets of stable transfectants were generated and those expressing levels of mIg comparable to the range normally seen on splenic B cells were studied. One set expressed an unmutated VH and an unmutated VL. The second set expressed the same pair of V regions except for a single somatic change in CDR3 of VH; this substitution increases the affinity of antibody for PC from 3 x 10(4) M-1 to 3 x 10(5) M-1. Ag:mIg interactions were assessed in the transfected cell lines by measuring calcium mobilization induced by stimulation with soluble PC Ag. As expected, the mutation that increased affinity for PC increased the sensitivity of transfectants to PC Ag. Relatively small changes in receptor number had a dramatic effect in the quantity and quality of a calcium response. Significantly, we found that Ag-specific signaling could occur with only a few thousand receptors per cell. Signaling differences were most noticeable with PC protein Ag (T-dependent form) compared with PC polysaccharide Ag (T-independent form). These results suggest that the down-regulation of mIg that follows B cell activation may have evolved to assist in the selection of B cell clones with higher affinity for Ag. Furthermore, the results also provide an explanation for why selection of higher affinity clones can occur with protein Ag but only poorly so with polymeric Ag. PMID- 8245441 TI - Naturally processed HLA class I bound peptides from c-myc-transfected cells reveal allele-specific motifs. AB - Naturally processed peptides, bound to HLA-A2, A68, B40 molecules, were isolated from a c-myc transfected lymphoblastoid B cell lines for sequence analysis. Forty three sequences of bound peptides could be grouped into three structural motifs. One of the peptide sequences obtained, SLLPAIVEL, was identical to a previously reported peptide bound to HLA-A2.1 and was used for grouping HLA-A2-bound peptides. A second motif, identical to that previously reported for HLA-A68-bound peptides, was also observed. A distinct third motif, consistent with the structure of the HLA-B40 "45 pocket," was observed. The peptides within this group contained glutamate in position 2, usually followed by a hydrophobic residue in positions 3 and 9. Within this motif group of peptides bound to MHC class I molecules, one peptide, HEETPPTTS, was 100% homologous to residues 243 251 of the c-myc protein. PMID- 8245442 TI - Naturally processed cytokine-derived peptide bound to HLA-class II molecules. AB - Sequence analysis of HLA-class II (HLA-DR beta 1-1502 and 1104)-bound self peptides from a transformed B cell line was performed. The sequences of naturally processed self-peptides bound to HLA-DR2 and DR5 were compared with protein and nucleic acid data bases for homology to known precursor proteins. Of the matches to known precursors, one peptide showed 100% homology to the third framework and CDR3 regions of Ig VH expressed by the line. Another peptide matched 100% to the human equivalent of macrophage inflammatory protein (MIP). A synthetic peptide corresponding to the naturally processed form of MIP (KPGVIFLTKRSRQV) was shown to inhibit Ag-specific HLA-DR beta 1*1104-restricted T cell proliferation. This indicates that the MIP peptide binds to HLA-DR beta 1*1104. The MIP peptide belongs to a set of peptides that showed uniform NH2-terminal processing. In this set, proline always occurred as the second residue followed by a basic lysine or arginine in position nine. This suggests that final NH2-terminal processing of peptides precedes their binding to MHC molecules. A distinct, second set of peptides showed ragged NH2-terminii, as has been reported for other naturally processed MHC-class II-bound self-peptides. PMID- 8245443 TI - Identification of a novel population of CD8 alpha+ beta- bone marrow-derived dendritic epidermal cells. This unique population is subsequently outnumbered by thymus-independent expansion of the CD8 alpha+ beta+ dendritic epidermal cells. AB - Maturation of some T cell populations has been suggested to occur in epithelial tissues. The CD8 molecule, which is expressed as heterodimers made of alpha/beta chains on virtually all peripheral T cells, is preferentially expressed as homodimers made only of alpha-chains on a subset of CD8+ mouse gut intraepithelial lymphocytes, which arise from bone marrow (BM) precursors in a thymus-independent mechanism. This unique population of CD8 alpha+ beta- T cells, however, has not been identified in other lymphoid tissues in normal adult mice. Because our previous study demonstrated that reconstitution of thymectomized irradiated mice with T cell-depleted BM cells resulted in the appearance of CD8+, TCR-alpha beta+ dendritic epidermal cells (DEC) and suggested that a proportion of the BM-derived DEC may mature within the epidermis, we asked whether the unique phenotype of CD8 alpha+ beta- could be detected among the CD8+ BM-derived DEC. In this report for the first time we identified this unique population of CD8 alpha+ beta- cells among the DEC. Although this population comprised the predominant fraction of the BM-derived DEC in the early post-transfer period (2 to 3 mo), gradual shift from the CD8 alpha+ beta+ phenotypes occurred during the late post-transfer period (4 to 6 mo) independently of the presence of the thymus. Kinetic studies on the BM-derived DEC revealed that this phenotypic shift could be caused by the subsequent expansion of the CD8 alpha+ beta+ DEC. In contrast, the CD8 alpha+ beta- population was the major subset of the BM-derived intraepithelial lymphocytes throughout the entire observation period and the phenotypic shift with age was never observed. These results indicate that CD8 alpha+ beta- cells are preferentially detected on T cells that home to the epithelial tissues and that in the epidermis, but not in the gut epithelia, the subsequent expansion of the CD8 alpha+ beta+ DEC would dilute the high number of the CD8 alpha+ beta- cells. PMID- 8245444 TI - Avidity for antigen can influence the helper dependence of CD8+ T lymphocytes. AB - To examine the influence of avidity on the helper dependence of CD8+ T cells, purified CD8+ T cells from transgenic mice expressing a TCR specific for H-2Kb were tested for their responsiveness to spleen cells expressing different densities of H-2Kb. High avidity interactions resulted in the induction of strong proliferative responses by purified CD8+ T cells. These cells synthesized their own IL-2 and IL-2R. By contrast, low avidity interactions failed to induce proliferation or detectable IL-2 synthesis, although IL-2R were induced and addition of rIL-2 induced a proliferative response. These results are consistent with a model in which TCR avidity for Ag can determine the helper dependence of a CD8+ T cell. PMID- 8245445 TI - Coexpression of NKR-P1 and alpha beta-TCR on lymphoid cells in fully xenogeneic (rat-->mouse) chimeras and syngeneically reconstituted (A-->A) rats. AB - We have identified three populations of cells among rat splenocytes expressing NKR-P1, including cells of NKR-P1bright/alpha beta-TCR-, NKR-P1dim/alpha beta TCR+, and NKR-P1dim/alpha beta-TCR- phenotypes. To study the phenotypic characteristics and development of these various cell populations, we have made use of transplanting untreated rat bone marrow into recipient rats (syngeneic) or mice (fully xenogeneic) conditioned with total body irradiation. Rat NK cells exhibit normal phenotypic markers (NKR-P1+, CD8+) and are normally functional by 28 days after reconstitution. We have found that the various populations of NKR P1+ cells are enriched significantly in the spleen and follow a characteristic pattern of development in the first mo after reconstitution. After syngeneic bone marrow reconstitution (rat-->rat), NKR-P1dim and NKR-P1bright cells (3-15%) can be demonstrated among splenocytes as early as day 3 after bone marrow transplantation. By day 7, the NKR-P1+ cells reach peak levels and comprise as much as 80% of splenic lymphoid cells, with 35% being NKR-P1bright and 45% being NKR-P1dim. The percentage of NKR-P1+ cells decreases over the next several wk until they constitute "normal rat" levels with 8 to 20% being NKR-P1bright and only 1 to 5% being NKR-P1dim. These same populations are also present in fully xenogeneic chimeras. In both models, approximately 80 to 90% of the NKR-P1dim cells were found to coexpress alpha beta-TCR at all time points. These NKR P1dim/alpha beta-TCR+ cells are not large granular lymphocytes and lack NK cell lytic activity against YAC-1 target cells. Additional analyses of cells derived from spleen, bone marrow, and thymus indicated that NKR-P1+ cells develop, for the most part, in a thymic-independent manner in our fully xenogeneic chimeras and syngeneically reconstituted rats. At present the developmental pathway of these NKR-P1dim cells remains speculative. PMID- 8245446 TI - Keratinocyte-derived IL-7 serves as a growth factor for dendritic epidermal T cells in mice. AB - Dendritic epidermal T cells (DETC) are CD3+, CD45+, CD4-, CD8-, TCR-V gamma 3/V delta 1+ T lymphocytes that reside in symbiosis with keratinocytes in mouse epidermis. To address mechanisms by which these cells survive and proliferate within the epidermal environment, we have tested the hypothesis that cytokines secreted by neighboring keratinocytes play relevant roles. The present study was conducted to determine whether keratinocytes produce biologically relevant amounts of IL-7, and, if so, to study its effects on DETC. The long term cultured DETC line, 7-17, and freshly isolated DETC exhibited dose- and time-dependent proliferative responses to rIL-7. These responses were blocked completely by anti IL-7 antibodies, whereas anti-IL-2 had no effect, indicating that DETC respond to IL-7 by an IL-2-independent mechanism. Proliferative responses depended on the state of cell activation; DETC stimulated 2 to 5 days earlier with Con A proliferated optimally to added IL-7, whereas cells stimulated 10 days earlier did not proliferate. DETC that failed to proliferate responded to IL-7 by showing prolonged survival; cells maintained for 7 days with IL-7 alone retained their capacity to proliferate in response to Con A. Mouse epidermal cells and Pam 212 keratinocyte line both expressed IL-7 mRNA constitutively, as demonstrated by reverse transcription-polymerase chain reaction analyses. The production of IL-7 by mouse keratinocytes was also confirmed; Pam 212 culture supernatants supported DETC proliferation, and this activity was diminished by 50% with added anti-IL-7 antibodies. These results indicate that keratinocytes produce IL-7 in biologically relevant amounts, which, in turn, serve to promote the survival and growth of DETC. IL-7-mediated communication between epithelial cells and gamma delta T cells may represent one mechanism to sustain the indefinite residence of gamma delta T cells in epithelial tissues of mice. PMID- 8245447 TI - Dissociation of the peptide-MHC class I complex limits the binding rate of exogenous peptide. AB - Soluble, single-chain molecules for two MHC class I alleles, H-2Kd and H-2Kb, were used to analyze the kinetics of antigenic peptide binding to MHC. After MHC preloading with radiolabeled or fluorescent peptides, the observed rate of MHC peptide complex dissociation increased after addition of an excess of unlabeled competitor peptide. Although exogenous peptides conforming to the allele-specific motif were required for the enhanced complex dissociation to occur, the dissociation rate of the complex was independent of exogenous peptide concentration. Similarly, the association rate of exogenous peptides was independent of concentration, reflecting the presence of low affinity peptides in the binding sites of the recombinant MHC proteins; the sequences of these endogenous peptides conform to the consensus motif for the MHC allele studied. Finally, the association rate of exogenous peptide decreased when MHC molecules were preloaded with high affinity peptides, and the binding of labeled high affinity peptide to isolated recombinant MHC was faster than the subsequent dissociation observed in the presence of competitor peptide. Taken together, these results imply that the rate of exogenous peptide binding is limited by the dissociation rate of the previously bound peptides. PMID- 8245448 TI - A lymphostromal molecule, thymic shared Ag-1, regulates early thymocyte development in fetal thymus organ culture. AB - Previously, we detailed the characterization of thymic shared Ag-1, a unique marker of immature thymocytes and isolated thymic stromal cells, defined by the mAb MTS 35. In this study, the functional relevance of this molecule to thymopoiesis was investigated by the addition of purified MTS 35 to fetal thymus organ culture. It down-regulated thymic shared Ag-1 expression and dramatically reduced thymocyte cell yield through inhibition of alpha beta-TcR+ T cell differentiation, post CD3-CD4-CD8- triple negative thymocytes. These effects were specific for the mAb MTS 35, because controls, which include both isotype-matched and other lymphostromal mAb, showed no similar effects. These results demonstrate that thymic shared Ag-1 is a functionally important marker of early thymocyte differentiation, particularly with regard to the alpha beta-TcR lineage. PMID- 8245449 TI - CD28-mediated costimulation is necessary for the activation of T cell receptor gamma delta+ T lymphocytes. AB - The role of costimulation in the activation of TCR-gamma delta cells in normal mice and mice transgenic (tg) for a TCR-gamma delta receptor was investigated. Activation of TCR-gamma delta cells required two signals. One signal was mediated by TCR occupancy, whereas a second signal was provided by accessory cells. The importance of the CD28/B7 interaction in the delivery of the second signal was demonstrated in multiple ways. First, addition of a soluble fusion protein homolog of CD28, CTLA4Ig, significantly inhibited the activation of G8 tg splenic TCR-gamma delta lymphocytes and intestinal epithelial TCR-gamma delta lymphocytes by Ag-bearing lymphocytes during primary stimulation. Similarly, both proliferation and IFN-gamma production were inhibited by addition of CTLA4Ig to secondary antigenic stimulation of G8 tg TCR-gamma delta cells. Second, an Ag bearing thymoma, EL-4, was only able to stimulate expanded G8 tg TCR-gamma delta cells when the thymoma expressed B7. This stimulation was blocked by both CTLA4Ig and anti-B7 antibody. Third, antibodies to CD28 were able to mimic the costimulatory affect of APC. TCR-gamma delta cells cultured with either Ag bearing fixed stimulator cells or submitogenic concentrations of immobilized anti pan TCR-gamma delta mAb proliferated only in the presence of anti-CD28 mAb. Finally, G8 tg cells produced IL-2 only in the presence of APC costimulation or anti-CD28 antibodies, and the addition of exogenous rIL-2 overcame the need for costimulation. Thus, autocrine IL-2 production is one of the major consequences of TCR-gamma delta cell costimulation. Together these data demonstrate that costimulation is necessary for the activation of TCR-gamma delta cells and can occur through CD28 interaction. PMID- 8245450 TI - Difference in signal transduction pathway for IL-2 and IL-4 production in T helper 1 and T helper 2 cell clones in response to anti-CD3. AB - To elucidate Th2 cell clone activation mechanism through TCR-CD3 complex, we examined the reactivity of Th2 cell clones to soluble anti-CD3 in the absence of accessory cells or costimulator. The soluble anti-CD3 stimulated IL-4 production of Th2 cell clones as efficiently as specific Ag. IL-4 production of Th2 cell clones was consistent with the elevation of intracellular free Ca2+ concentration ([Ca2+]i). The elevation was slow and sustained but occurred consistently after the anti-CD3 stimulation in all Th2 cell clones tested. The [Ca2+]i elevation appeared to depend on Ca2+ influx because it could not be observed in Ca(2+)-free medium. Several chemicals such as cholera toxin, neomycin, and herbimycin A, which have been shown to block phosphatidylinositol-4,5-bisphosphate (PIP2) breakdown pathway or protein tyrosine kinase activation, exerted no effect on the IL-4 production. In accordance with these findings, neither PIP2 breakdown nor protein tyrosine phosphorylation was observed in Th2 cell clones stimulated with anti-CD3. The inclusion of anti-CD4 in culture and the depletion of protein kinase C (PKC) did not affect IL-4 production of Th2 cell clones either. These findings support a hypothesis that Th2 cell clones use a signaling pathway for IL 4 production that is independent of protein tyrosine kinase, PIP2 breakdown or PKC, and that the [Ca2+]i elevation is the only pathway common to an IL-2 production of Th1 cell subset. PMID- 8245451 TI - Two separate mechanisms of T cell clonal anergy to Mls-1. AB - T cell tolerance to superantigen can be mediated by clonal anergy in which Ag specific mature T cells are physically present but are not able to mount an immune response. We induced T cell unresponsiveness to minor lymphocyte stimulations locus antigen (Mls)-1a in mice transgenic for TCR V beta 8.1 in three different systems: 1) injection of Mls-1a spleen cells, 2) mating with Mls 1a mice, and 3) bone marrow (BM) chimeras in which Mls-1a is present only on nonhematopoietic cells. CD4+8-V beta 8.1+ cells from all these groups did not proliferate in response to irradiated spleen cells from Mls-1a mice. We compared the response of these cells by T cell/stimulator cell conjugate formation, Ca2+ mobilization, and proliferation assays. The mechanisms underlying the unresponsiveness of these T cells appear to differ. CD4+8-V beta 8.1+ cells from Mls-1a spleen cell-injected mice mobilized cytoplasmic Ca2+ but proliferated at a reduced level in response to cross-linking with anti-TCR mAb. However, these cells formed conjugates, mobilized Ca2+, and proliferated in response to Mls-1a when activated B cells were used as stimulators, although they produced reduced levels of IL-2. In Mls-1a/b V beta 8.1 transgenic mice, a subset in CD4+8-V beta 8.1+ cells did not mobilize cytoplasmic Ca2+ after TCR cross-linking. Their conjugate formation, Ca2+ mobilization, or proliferation in response to Mls-1a on activated B cells was undetectable. Finally, CD4+8-V beta 8.1+ cells from the BM chimeras proliferated to TCR cross-linking at a partially reduced level and formed conjugates, mobilized Ca2+, and proliferated in response to Mls-1a on activated B cells. These features suggest that the mechanisms underlying the maintenance of anergy in Mls-1a spleen cell-injected mice are distinct from those in Mls-1a mice. PMID- 8245452 TI - A cDNA clone encoding the mouse Qa-1a histocompatibility antigen and proposed structure of the putative peptide binding site. AB - We have isolated a cDNA clone which encodes the Qa-1a histocompatibility Ag from a library prepared from Con A-activated B10.BR mouse spleen cells. The clone encodes a protein of 322 amino acids with three potential N-glycosylation sites. The coding sequence shows strongest similarity with that of the T23d gene of DBA/2 mice which encodes the Qa-1b molecule. Molecular modeling of the putative peptide-combining site indicates most of the differences between Qa-1a and Qa-1b are located peripheral to the binding cleft, with only two amino acid substitutions, at positions 9 and 24, which might affect peptide binding. Many features of the Qa-1 binding cleft are also conserved in the rat RTBM.1 and in human HLA-E molecules. This suggests that all of these molecules may associate with structurally similar peptides. PMID- 8245453 TI - E-box activator of the C4 promoter is related to but distinct from the transcription factor upstream stimulating factor. AB - The murine C4 promoter contains a motif (E-C4) active in transcriptional activation whose structure complies with the E-box consensus sequence recognized by the helix-loop-helix transcription factors. This site is found also in human and rat C4 promoters and has the structure (-75) CACGTG (-70) characteristic of the class B subset of b-helix-loop-helix-zipper proteins. We have challenged the hypothesis that the protein factor responsible for the E-C4-mediated transcriptional activation is identical to one of the previously characterized nuclear factors. The molecular mass of the E-C4 factor is slightly bigger than that of Hela upstream stimulating factor (USF) (43/44 kDa). Moreover, the nucleotides immediately adjoining the E-C4 core sequence contribute to the distinctive fine specificity of the E-C4 factor. Optimized USF and MYC DNA binding sites, which differ in the nucleotides bordering the hexanucleotide box displace the E-C4 factor in competition assays but with lesser efficiency than the E-C4 site itself. Finally, the E-C4 factor fails to exhibit the heat resistance characteristic of USF proteins. The results show that the E-C4 transcription factor has DNA binding properties overlapping those of other helix loop-helix proteins but is structurally distinct from the factors so far described. Although C4 is not the first liver gene endowed with an E-box-mediated activation, it affords the first example where such activation takes place in the context of a TATA-less promoter, and is functionally linked to an initiator element-dependent transcription. PMID- 8245454 TI - Few V gene segments dominate the T cell receptor beta-chain repertoire of the human thymus. AB - We undertook a large sample census of the TCR-beta repertoire from a single human thymus (300 clones from an unamplified cDNA library), to establish the statistics of the rearranged V, D, J, and C gene segments. The assortments of all germ-line segments are subject to significant biases and thus critically reduce the effective germ-line contribution to beta-chain diversity. Thirty-two V genes characterize the whole sample, but surprisingly as few as seven genes from different families encompass half of the beta-chain repertoire. Furthermore, a Spearman rank order correlation test of the thymic V beta frequencies with those inferred in other studies using RNase protection assays shows a statistically significant similarity. Thus, in the establishment of thymic V beta frequencies in man, rearrangement preferences intrinsic to the progenitors of TCR-beta expressing cells override HLA- and Ag-dependent biasing factors. By implication, a large enough pool of direct progenitor cells (maybe as high as thousands) must exist to secure V beta frequencies against large random fluctuations. Uncorrelated with the V gene bias, the representation of D beta and J beta segments is also far from even. Notably, D beta 1 and J beta 2.1 and 1.2 predominate. Using the above mentioned rank order statistic, we also find that unlike V genes, the bias in the J segment usage carries over into the frequencies of peripheral T lymphocyte populations. PMID- 8245455 TI - Third component of trout complement. cDNA cloning and conservation of functional sites. AB - Of the 30 distinct complement proteins recognized to date, C3 is probably the most versatile and multifunctional molecule known, interacting with at least 20 different proteins. It plays a critical role in both pathways of complement activation and participates in phagocytic and immunoregulatory processes. Structural and functional analysis of C3 from different species, in addition to phylogenetic information, provides insights into the structural elements mediating the various functions. This study describes the cDNA cloning of one of two isoforms of the third complement component, C3-1, of rainbow trout (Salmo gairdneri) and the analysis of its functional sites. By screening a trout liver lambda gt11 library with anti-trout C3 chain-specific antibodies and polymerase chain reaction we have determined the cDNA sequence of trout C3-1. The obtained sequence is in complete agreement with the protein sequence of several tryptic peptides, corresponding to different regions of trout C3-1. C3-1 consists of 1640 amino acids with a calculated molecular mass of 181,497 Da. The sequence contains two potential N-glycosylation sites, one on each chain of C3. The deduced protein sequence showed 44.1, 43.3, 44.2, 44.9, 43.1, 43.8, 45.9, 29.9, and 33.1% amino acid identities to human, mouse rat, guinea pig, rabbit, cobra, frog, hagfish, and lamprey C3, whereas the identities to human C4, C5, and alpha 2M are 30.4, 28, and 22.9%, respectively. The trout C3 amino acid sequence shows clusters of high and low similarity to C3 from other species. In the regions of high similarity belong the C3 domains that contain the thiolester site and the properdin binding sites, whereas the regions that correspond to regions of human C3 where CR1 and CR2 bind show low amino acid sequence similarity. The deduced amino acid sequence shows that the C3 convertase cleavage site (Arg-Ser) is conserved in trout C3, whereas the factor I cleavage sites are Arg-Ala and Arg Thr instead of Arg-Ser, which is found in the C3 of other species. Protein sequencing of the trout C3 fragments fixed on zymosan during complement activation confirmed the cleavage of trout C3 by trout C3 convertase and factor I at Arg-Ser and Arg-Thr, respectively. PMID- 8245456 TI - cAMP activates the IL-5 promoter synergistically with phorbol ester through the signaling pathway involving protein kinase A in mouse thymoma line EL-4. AB - Expression of the IL-5 gene in T cells is induced in response to Ag stimulation; however, functional analysis of the IL-5 gene has been limited by lack of an appropriate transfection assay to facilitate measurement of the IL-5 promoter activity in response to T cell activation signals. Here, we describe a transient transfection system with which the IL-5 promoter activity can be assayed quantitatively. Using mouse thymoma line EL-4 cells, which produce several lymphokines including IL-2, IL-3, IL-4, IL-10, and GM-CSF in response to PMA, the effect of cAMP on IL-5 production was examined. These cells produce a low level of IL-5 when stimulated with PMA alone; however, N6, O2-dibutyryl cAMP (Bt2cAMP), in combination with PMA, augmented by more than tenfold the IL-5 production at the mRNA and the protein levels. Likewise, a transient transfection assay revealed that Bt2cAMP activated the IL-5 promoter more than tenfold, in a PMA dependent manner, thereby indicating that two signals, PMA and cAMP, are required for optimal activation of the IL-5 promoter. Activation of the IL-5 promoter in response to Bt2cAMP and PMA depends on the region spanning from nucleotide position -1,200 to +33 relative to the transcription initiation site. Action of cAMP on the IL-5 promoter is mimicked by cotransfection of the expression plasmid containing cDNA encoding the catalytic subunit of protein kinase A, hence, cAMP probably exerts its action through the signaling pathway that involves protein kinase A. In contrast, Bt2cAMP almost completely inhibited the PMA-dependent activation of the endogenous IL-2 gene as well as the transfected IL-2 promoter. These results indicate that the IL-5 gene in EL-4 cells is positively regulated by cAMP in a manner opposite that for the IL-2 gene. PMID- 8245457 TI - Role of the B cell antigen receptor in antigen processing and presentation. Involvement of the transmembrane region in intracellular trafficking of receptor/ligand complexes. AB - To study the role of the B cell Ag receptor (membrane-bound Ig [mIg]) in Ag processing and presentation, we have generated several Ig transfectants that express transfected TEPC-15 idiotype (T15-Id) mIgM with phosphorylcholine (PC) binding specificity. The wild-type Ig transfectant is able to present a specific Ag (e.g., PC-conjugated hen egg-white lysozyme [PC-HEL]) more efficiently than a nonspecific Ag (HEL) to a T cell hybridoma recognizing an epitope on the HEL molecule. A substitution in the entire spacer, transmembrane, and cytoplasmic region of mIg with an equivalent region of I-A alpha chain completely abolishes this mIg-enhanced Ag presentation. Experiments with the wild-type and substituted variant Ig transfectants suggest that this substitution may interfere with normal intracellular trafficking of mIg after cross-linking with specific Ag or antibodies specific for the mIg (anti-T15-Id mAb). Prolonged treatment of the wild-type Ig transfectants with specific Ag or anti-T15-Id mAb reduces the surface expression of T15-Id mIgM and leads to an accumulation of T15-Id mIgM inside the cells. The reduced surface expression and the elevated cytoplasmic accumulation of T15-Id mIgM are not observed in the variant Ig transfectant. Despite the ability of the variant to endocytose ligands similarly to the wild type Ig transfectant, this variant displays a higher rate of recycling of the mIg/ligand complexes back to the cell surface and a lower rate of degradation of the ligands. These abnormalities may be responsible for the deficiency in mIg mediated enhancement of Ag presentation in the variant Ig transfectant. Therefore, our results suggest that the transmembrane region of mIg is involved in intracellular trafficking of receptor/ligand complexes and that proper delivery and handling of internalized Ag are required for the enhanced presentation of specific Ag by B cells. PMID- 8245458 TI - Analysis of T cell receptors specific for recognition of class IB antigens. AB - T cells that recognize a peptide presented by a self-class IA molecule generally use a restricted repertoire of V beta and V alpha receptors. In contrast, alloreactive T cells, which recognize alloantigens that present a wide array of peptides, use a diverse repertoire, particularly in the CDR3 loop. Because the T cell repertoire directed against class IB alloantigens is not known, we examined V-D-J sequences in V beta chains specific for Qa-1 and similar sequences in both V beta and V alpha chains specific for Qa-2. We observed that 14 Qa-1-specific clones use a limited number of V beta segments and 8 of 14 express V beta 8.2 and have a conservation of charged residues in the CDR3 loop, particularly between residues 99 and 101. Thirteen of the 14 clones rearrange to the second J beta cluster and use within this cluster is restricted. Alloreactive anti-Qa-1 T cells can be assigned into three different specificity groups based on a Qa-1 modifying gene, Qdm, as well as Qa-1 epitope expression on Tap-2-deficient RMA-S cells. Receptors from members of each specificity group are more similar in their CDR3 loop to each other than members of the other groups. These data lend support to the Qa-1 class IB Ag presenting a limited number of peptides to T cells or in some manner limiting the development of a diverse alpha beta T cell repertoire. The alpha- and beta-chains from nine alloreactive anti-Qa-2 clones were analyzed. V beta use was limited to use of V beta 7 or a member of the V beta 8 family. Rearrangements were solely to the second J beta cluster. The use of V alpha and J alpha segments were diverse. Although conserved residues or motifs were observed in the CDR3 regions of both the beta- and alpha-chains, the extent of conservation was less than that for anti-Qa-1 receptors. Anti-Qa-2 T cells can be divided into two specificities, Q6 and Q7. No common features were apparent between these groups. PMID- 8245459 TI - Characterization of the gene for the human high affinity IgE receptor (Fc epsilon RI) alpha-chain. AB - The Fc epsilon RI couples the mast cell-surface binding of IgE and Ag to a complex series of intracellular events culminating in cell activation and degranulation. The alpha-chain of Fc epsilon RI constitutes the Ig-binding subunit of this heterotetrameric receptor, and is itself a member of the Ig gene superfamily. We have isolated a human genomic DNA clone containing the entire Fc epsilon RI alpha gene, and completely sequenced a region from 1257 bp 5' of the transcription start site, to 513 bp 3' of the last exon of the gene. As with the previously characterized rat and mouse genes, human Fc epsilon RI alpha consists of five exons and four introns, and spans 5889 bp of genomic DNA. The splice donor and acceptor sites deduced by comparison with the cDNA sequence corresponded exactly to the locations found in analogous rodent genes. By mapping the 5' end of Fc epsilon RI alpha transcripts we found three major transcription initiation sites 24, 27, and 29 bp upstream of the ATG translation initiation codon. As well, several longer minor transcripts were seen, with a maximum of 60 nt of 5'-untranslated sequence. About 650 bp of DNA upstream of the ATG translation initiation codon were compared among human, rat, and mouse Fc epsilon RI alpha sequences in search of common motifs that might mediate conserved regulatory interactions with DNA binding proteins. A 172-bp region of the human Fc epsilon RI alpha 5'-flanking sequence was highly conserved in both rodent species. Further studies will be required to determine whether these or other sequences are involved in Fc epsilon RI alpha gene regulation. PMID- 8245460 TI - Selection for amino acid sequence and J beta element usage in the beta chain of DBA/2V beta b- and DBA/2V beta a-derived myoglobin-specific T cell clones. AB - Previous experiments in our laboratory have demonstrated that there is a marked restriction on the TCR beta-chain usage in DBA/2 mice in response to the sperm whale myoglobin (SpWMb) determinant 110-121, predominated by the use of the V beta 8.2 gene element. We analyzed the response of mice that had been genetically depleted of V beta 8+ T cells by generating a DBA/2 line that carries the V beta a TCR haplotype. Despite the very limited TCR repertoire expressed by DBA/2V beta a mice, they made an excellent response after immunization with the SpWMb 110-121 peptide. Data presented in this manuscript demonstrate that there is an equally restricted TCR V beta-chain utilization in the T-cell response to the determinant SpWMb 110-121 in DBA/2V beta a mice. Unexpectedly, there was a shift of MHC restriction of this determinant to T cells in the V beta a strain when compared with the V beta b strain of DBA/2 mice. We had previously demonstrated that DBA/2 mice utilized both the hybrid E alpha dA beta d MHC molecule as well as the conventional A alpha dA beta d molecule as presenting elements in response to SpWMb 110-121. Data presented in this manuscript demonstrate that the T-cell response in DBA/2V beta a mice is entirely restricted by the A alpha dA beta d MHC class II molecule. By analyzing a panel of SpWMb 110-121-specific T-cell clones from DBA/2V beta a mice, we were able to study the TCR repertoire expressed on T cells from mice that lack the V beta 8.2 gene. The V beta usage by the panel of clones analyzed was remarkably homogeneous. Thirteen of the 17 clones analyzed used the V beta 1 gene segment. Perhaps more striking was the junctional region nucleotide and amino acid sequences that were shared among these clones and that were similar to the V beta 8.2 clones analyzed previously. All clones assayed used the J beta 2.6 element, as did the great majority of the V beta 8.2 clones analyzed from DBA/2 (and B10.D2) V beta b mice. Importantly, in each strain of mice, irrespective of the V beta utilized, each TCR appeared to have selected an acidic amino acid in the beta-chain at position 100.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8245461 TI - Met-ase: cloning and distinct chromosomal location of a serine protease preferentially expressed in human natural killer cells. AB - A cDNA clone encoding a human NK serine protease was obtained by screening a lambda-gt10 library from the Lopez NK leukemia with the rat natural killer Met ase (RNK-Met-1) cDNA clone. In Northern blot analysis human Met-ase (Hu-Met-1) cDNA hybridized with a 0.9-kb mRNA in two human NK leukemia cell lines, unstimulated human PBMC, and untreated purified CD3-CD56+ large granular lymphocytes. Unlike other members of the granzyme family that are highly expressed in activated peripheral T cells, the Hu-Met-1 transcript was barely detected in a population of PMA and ionomycin or IL-2-treated high density T cells. Several in vitro cultured Burkitt lymphomas, chronic- and promyeloid leukemias, acute lymphoblastic leukemias, and colon and ovarian carcinomas and colon and ovarian carcinomas did not express Hu-Met-1 mRNA. Hu-Met-1 mRNA expression in a small number of human T cell tumor lines did not correlate with any particular phenotype or stage of development. The presence of Hu-Met-1 mRNA closely correlated with the Met-ase activity of cellular lysates prepared from these various human peripheral blood subsets and in vitro cultured cell lines. Met-ase activity detected in whole cell lysates of cytotoxic lymphocytes was associated with the cytoplasmic granules of these cells. The nucleotide sequence of the Hu-Met-1 cDNA clone encodes a predicted serine protease of 257 amino acids. The predicted protein is an active enzyme of 232 amino acids with a calculated unglycosylated m.w. of 27,100. Hu-Met-1 is 66% identical to RNK-Met-1 at the amino acid level. The human and rat mature protein sequences conserve the active site His, Asp, and Ser amino acids that form the catalytic triad of serine proteases, all 8 cysteine residues, and several amino acids critical in the formation of the substrate binding pocket. The gene for the Hu-Met-1 serine protease is located on chromosome 19, which distinguishes it from any other member of the human granzyme family. PMID- 8245462 TI - Binding site on the murine IFN-gamma receptor for IFN-gamma has been identified using the synthetic peptide approach. AB - We have studied the structural parameters involved in the binding of murine IFN gamma (MuIFN-gamma) to its receptor. Ten synthetic overlapping peptides corresponding to the extracellular domain of the MuIFN-gamma receptor (MuIFN gamma R) were synthesized. In direct binding studies, biotinylated MuIFN-gamma bound specifically to receptor peptide (95-120). Further, the NH2-terminal IFN gamma peptide, MuIFN-gamma (1-39), also specifically bound to receptor peptide (95-120). Binding of both labeled MuIFN-gamma and MuIFN-gamma (1-39) to MuIFN gamma R peptide (95-120) was inhibited by either unlabeled molecule. The COOH terminal receptor binding peptide, MuIFN-gamma (95-133), neither bound to any receptor peptides nor blocked the binding of intact MuIFN-gamma or MuIFN-gamma (1 39) to receptor peptide (95-120). Polyclonal antibodies to each of the peptides were then produced. Each of the anti-peptide antisera recognized its corresponding peptide and bound denatured cloned soluble receptor by Western blotting. Furthermore, the antisera to peptides representing the inner region of the extracellular domain of the receptor bound to nondenatured soluble MuIFN gamma R. Specifically, antisera to the receptor peptides (73-97), (95-120), (118 143), (142-163), and (161-182) bound to soluble MuIFN-gamma R, whereas antisera to peptides (1-21), (20-49), (46-74), (178-203), and (202-227) did not bind. Most important, antisera to peptides (95-120) and (118-143) competed with [125I]MuIFN gamma for binding to soluble receptor. These results show that the region of the MuIFN-gamma R encompassing amino acid residues (95-120) is a binding site on the receptor for the NH2-terminal of MuIFN-gamma by direct binding, and that the larger region (95-143) on the receptor may play a role in binding of intact MuIFN gamma based on blocking of binding by site-specific antibodies. PMID- 8245463 TI - Structure of the gene for the F allele of complement receptor type 1 and sequence of the coding region unique to the S allele. AB - The genes for human complement receptor type 1 (CR1) F and S alleles have been cloned and span a region of 133-160 kb on chromosome 1. The F allele was found to comprise 39 exons and the S allele contains an additional 8 exons. The leader sequence and 5'-untranslated region are contained in one exon. Each of the long homologous repeats (LHR), which contain seven short consensus repeats (SCR), is composed of 8 exons. Within a LHR, SCR 1, 5, and 7 are each encoded by a single exon, SCR 2 and 6 are each encoded by 2 exons, and a single exon codes for SCR 3 and 4. The transmembrane region is encoded by 2 exons and the cytoplasmic domain and the 3'-untranslated regions are coded for by separate exons. The sequences of the eight S allele-specific exons were very similar to those from LHR-A and -B, as was predicted by comparison of the genomic restriction maps. It had previously been suggested that the alleles of CR1 have arisen by a mechanism of unequal crossover. A comparison of intron sequences from LHR-A, -B, -C, and -S revealed data that support this hypothesis. The homologies of intron sequences from LHR-A, -B, and -C indicated that the crossover event between LHR-A and -C that gave rise to LHR-B probably occurred within the fourth exon of these LHR. Likewise, the crossover event between LHR-A and -B that produced LHR-S probably occurred within a 383 bp region around the sixth exon. Analysis of RNA from peripheral blood cells by the S1 nuclease assay indicated that the transcription start site is 111 bp upstream of the translation initiation codon ATG. The 5' rapid amplification of cDNA ends confirmed this position as a transcription start site and revealed another possible start site 29 bp further upstream. PMID- 8245464 TI - Direct evidence that interferon-beta mediates enhanced HLA-class I expression in measles virus-infected cells. AB - Viral infection results in enhancement of HLA-class I expression in a number of cell types, including glial cells, which normally do not express these molecules. This enhancement may occur through a direct interaction between a viral component and the HLA-class I gene or indirectly through virus-induced soluble factors produced by infected cells. These include cytokines such as IFN-gamma, IFN alpha/beta, and TNF-alpha, known to enhance class I expression. Measles virus (MV) infection of a human glioma cell line (U-105 MG) and of primary human umbilical vein endothelial cells enhances the expression of HLA-class I molecules on these cells. The enhancement of HLA-class I is dependent on infectious virus, as antibody-neutralized MV has no effect on class I expression. In this study, we demonstrate the presence of an HLA-class I-enhancing factor in supernatants from MV-infected cells. The supernatant class I-enhancing factor is not IFN-gamma, IFN alpha, or TNF-alpha because MV-infected cells did not produce measurable levels of these cytokines as detected by immunoassay or polymerase chain reaction. In contrast, IFN-beta is produced by the infected cells and the supernatant class I enhancing factor could be entirely neutralized by antibodies to IFN-beta, but not antibodies to IFN-alpha, TNF-alpha, or non-immune sera. Furthermore, preincubation of cells with neutralizing antibodies to IFN-beta prior to infection blocked MV enhancement of HLA-class I completely in the U-105 MG cells and by as much as 74% in the umbilical vein endothelial cells. The results of these experiments provide direct evidence that enhanced HLA-class I expression in MV-infected cells is mediated primarily by IFN-beta. PMID- 8245465 TI - Conglutinin acts as an opsonin for influenza A viruses. AB - Since the 1940's, non-Ig inhibitors of influenza A virus (IAV) hemagglutination activity and infectivity have been recognized in mammalian sera. Recently, the heat labile (beta) inhibitor of this type was identified by indirect methods as the lectin, conglutinin. In support of this hypothesis, we found that purified conglutinin strongly inhibited hemagglutination activity and infectivity of IAV. By using IAV strains with specific variations in glycosylation of the hemagglutinin molecule, we showed these effects to be mediated by binding of conglutinin to high mannose carbohydrate attachments on the viral hemagglutinin. Through the same mechanism conglutinin caused aggregation of IAV particles. Human neutrophils produce hydrogen peroxide upon exposure to IAV. Also, after a brief period of exposure to IAV, neutrophils exhibit depressed responsiveness (deactivation) upon exposure to other stimuli (e.g., chemotactic peptides). These phenomena may be related to the in vivo inflammatory response during IAV infection, and to the propensity of IAV-infected subjects to suffer bacterial superinfection. Pre-incubation of IAV with conglutinin markedly potentiated human neutrophil hydrogen peroxide production in response to the virus. This effect correlated with the ability of conglutinin to aggregate the virus. IAV treated with conglutinin also caused significantly less neutrophil deactivation than did the unopsonized virus. These enhancements of neutrophil respiratory burst responses by conglutinin were again mediated by binding of the lectin to viral carbohydrates. The mammalian C-type lectin family includes conglutinin, mannose binding protein, and surfactant proteins A and D. These lectins may be important constituents of the initial host response to IAV, by inhibiting IAV infectivity directly, causing viral aggregation, and acting as opsonins to enhance phagocyte responses to the virus. PMID- 8245466 TI - Very late antigen-5 and complement receptor type 3 cooperatively mediate the interaction between Bordetella pertussis and human monocytes. AB - Nonopsonized Bordetella pertussis, the causative agent of whooping cough, can attach to and become ingested by human monocytes. It has been reported that complement receptor type 3 (CR3) on human monocyte-derived macrophages binds filamentous hemagglutinin expressed on B. pertussis. In the present study, the role of very late antigen-5 (VLA-5) in the attachment of B. pertussis to adherent human monocytes was investigated. It was found that soluble fibronectin and soluble mAb against VLA-5 markedly inhibited the attachment of B. pertussis to monocytes. When VLA-5 on monocytes was cross-linked by plating these cells onto surfaces precoated with fibronectin or mAb against VLA-5, the binding of both B. pertussis and C3bi-coated sheep erythrocytes to these cells was significantly enhanced, whereas the binding of a B. pertussis mutant strain deficient in filamentous hemagglutinin was not affected. The enhanced attachment of B. pertussis to monocytes plated onto fibronectin-coated surfaces was markedly inhibited by soluble mAb against CR3. Neutrophils, which express similar levels of CR3 and about 10-fold lower levels of VLA-5 as compared with monocytes, did not bind B. pertussis. Together, these results indicate that VLA-5 is involved in the attachment of B. pertussis to monocytes and that cross-linking of VLA-5 enhances the attachment of B. pertussis to monocytes by augmenting the binding activity of CR3. We propose that the attachment of B. pertussis to monocytes occurs in two steps: binding and cross-linking of VLA-5 by B. pertussis enhances the binding activity of CR3, which in turn facilitates the subsequent binding of these bacteria to the latter receptor. PMID- 8245467 TI - Beta 2M-/- knockout mice contain low levels of CD8+ cytotoxic T lymphocyte that mediate specific tumor rejection. AB - C57BL/6 mice with a disrupted beta 2M gene (beta 2M-/- mice) express very low levels of MHC class I molecules and are deficient for CD8+ T lymphocytes. Because CD8+ T cells are thought to be a principle effector cell in tumor rejection, we have assessed the ability of beta 2M-/- mice to respond to tumors. beta 2M-/- knockout mice were challenged with seven independent MHC allogeneic and syngeneic tumors. The beta 2M-/- mice responded very similarly to their CD8+ beta 2M+/- littermates in that they rejected high dose challenges of 4/5 allogeneic tumors and were susceptible to 3/3 syngeneic tumors. In vivo depletion of CD4+ or CD8+ cells from the beta 2M-/- mice resulted in susceptibility to allogeneic tumor. The apparent requirement for CD8+ cells for tumor immunity was corroborated by in vitro assays in which depletion of CD8+ but not CD4+ T cells eliminated tumor specific CTL activity. mAb blocking studies in which target tumor cells were incubated with MHC class I-specific mAb demonstrated that the tumor-specific CD8+ activity was MHC class I restricted. beta 2M-/- mice therefore contain very small quantities of potent, CD8+ T cells that are capable of rejecting large challenges of allogeneic tumor cells. PMID- 8245468 TI - In vitro candidastatic properties of the human neutrophil calprotectin complex. AB - Microbiostatic mechanisms may contribute substantially to host defense against infection by certain microbes. We studied the candidastatic activity of human neutrophils, neutrophil cytosol, and neutrophil-derived "calprotectin," a cytosolic protein complex comprised of two subunits, MRP8 and MRP14. Intact neutrophils, neutrophil lysates (prepared by ultrasonic disruption, freezing and thawing, or nonionic detergent extraction), and granule-depleted neutrophil cytosol were effective in restricting the growth of Candida albicans in a nutrient-rich tissue culture medium, RPMI 1640. Neither a subcellular fraction enriched in neutrophil granules nor selected purified granule components (lactoferrin, myeloperoxidase, cathepsin G, leukocyte elastase, lysozyme, and defensins) exerted candidastatic activity in this medium. Gel filtration liquid chromatography, anion exchange FPLC, and SDS-PAGE showed that the fungistatic factor in neutrophil cytosol was associated with the calprotectin complex. Its antifungal effects included restriction of yeast phase and mycelial growth and inhibition of glucose incorporation by yeast phase cultures. The antifungal effects of calprotectin were sustained for over 120 h and were inhibited by zinc. However, studies with 65Zn-enriched RPMI suggested that the candidastatic effects of calprotectin were not mediated by sequestration or binding of zinc. After reversed phase HPLC, calprotectin fractions containing MRP14 exhibited fungistatic activity, whereas fractions depleted of MRP14 but enriched for MRP8 lacked fungistatic activity. The results support a potentially significant role for the calprotectin complex of neutrophil cytosol in antifungal defense and suggest that MRP14 is of key importance in that activity. PMID- 8245469 TI - Shared T cell-defined antigens on independently derived tumors. AB - We report that a subset of tumors independently derived from a cloned line of contact-inhibited, non-tumorigenic murine fetal fibroblasts confer cross protective immunity against each other in vivo. Concordant with the in vivo cross protection, cytolytic T cell clones from mice immunized with one of these tumor lines specifically lyse the three other lines in the same set but do not cross react with either the nontumorigenic parental line or another similarly derived tumor line representing a different antigenic profile. This and other recent evidence for shared expression of tumor rejection Ag contrasts with the antigenic diversity previously described for chemical- and radiation-induced tumors. In the interpretation of such data it is essential to distinguish between Ag expressed in association with the transformation process and Ag induced by random mutation of already transformed cells. PMID- 8245470 TI - Antigen-dependent stimulation by bone marrow-derived mast cells of MHC class II restricted T cell hybridoma. AB - This paper describes a new role for mast cells as being able to present Ag to immune T cells. A mouse bone marrow-derived mast cell population obtained after 3 wk of culture in a conditioned medium has been shown to express a variety of membrane-associated Ag, including MHC class II and class I Ag, CD23, CD32, high affinity receptor for IgE, and CD4. Expression of MHC class II molecules was up regulated upon stimulation with LPS but not with IFN-gamma and was down-regulated after exposure of mast cells to IL-3 treatment. We have demonstrated that mast cells were able to present native Ag as well as immunogenic peptides to MHC class II-restricted T cell hybridoma. The inhibition of Ag presentation after mast cells have been treated with ammonia suggests that Ag catabolism in intracytoplasmic compartment as a key step in Ag handling takes place in these cells. The MHC class II molecule is the restricting element for the presentation of OVA and the lambda repressor from bacteriophage lambda to a panel of specific T cell hybridomas, as demonstrated by the blocking effect of anti-MHC class II mAb on the Ag-presenting function. A characteristic feature of mast cells is the generation of a narrower immunogenic peptide repertoire as compared with A20 and LBB 3.4.16, a B lymphoma cell line, and a B cell hybridoma, respectively. This novel function of mast cells brings to a much closer connection inflammatory and immunologic processes and sheds new light on the biology of mast cells and particularly on the specific allergic responses. PMID- 8245471 TI - IL-3 alters free arachidonic acid generation in C5a-stimulated human basophils. AB - IL-3 is known to enhance the secretion of several mediators from human basophils activated by receptor-mediated stimuli. IL-3 can cause a qualitative change in the mediator release pattern for C5a-mediated stimulation; without IL-3, C5a causes no leukotriene release whereas in the presence of IL-3, significant leukotriene occurs. This study examines the influence of a 15-min pretreatment of basophils with IL-3 (10 ng/ml) on several signal transduction events. Basophils stimulated with C5a typically displayed only a transient cytosolic Ca2+ response [Ca2+]i, attributed to the release of intracellular calcium stores. IL-3 had sporadic, statistically nonsignificant, effects on the peak of this initial response as well as inconsistent effects on the generation of a second phase in the [Ca2+]i response (i.e., that which is caused by the influx of extracellular Ca2+). IL-3 also had no effect on resting [Ca2+]i levels. Challenge of basophils in the presence of EGTA had little effect on the amount of leukotrienes generated. This maneuver did not influence the initial transient elevation of [Ca2+]i. Although basophil leukotriene release is usually slow, after exposure to IL-3 it was found that leukotriene release occurred rapidly, during the brief window of time (0 to 45 s) in which the [Ca2+]i transient occurred. These results suggested that the generation of AA was accelerated by pretreatment with IL-3. Subsequent mass measurements of free AA by gas chromatography-mass spectroscopy showed: 1) IL-3 itself caused no free AA generation; 2) without IL-3, C5a stimulated little or no free AA generation; and 3) in the presence of IL-3, C5a generated substantial levels of free AA at an accelerated rate. A similar acceleration in the rate of free AA generation, without any apparent increase in the amount, occurred in IL-3-primed basophils stimulated with FMLP. Additional studies showed that IL-3 had no effects on whole cell ATP levels and that the kinase inhibitor, staurosporine, did not inhibit the ability of IL-3 to cause enhanced responses to C5a. We conclude that the primary effect of a short period of pretreatment with IL-3 is to couple the generation of free AA to C5a-mediated stimulation in particular and to accelerate its generation after other stimuli. Enhancements of the [Ca2+]i response, while sporadic and at best modest, may have some influence on enhanced mediator release but did not clearly explain the functional effects of IL-3. PMID- 8245472 TI - Tyrosine phosphorylation of a gamma-chain homodimer associated with Fc gamma RIII (CD16) in cultured human monocytes. AB - The efficient expression of transmembrane-anchored Fc gamma RIIIa requires the presence of other peptides, such as the gamma-chain of the IgE receptor I or the zeta-chain of the TCR. We found that Fc gamma RIIIa in cultured human monocytes is specifically associated with the gamma-chain homodimer, and that the gamma chains in this complex are phosphorylated on tyrosine residues. Anti-Fc gamma RIII immunoprecipitates, which were prepared from 1% digitonin lysates of cultured human monocytes, incorporated phosphate into a homodimer consisting of two 14-kDa polypeptides when incubated with [gamma-32P]ATP. Identity of this co associated structure of Fc gamma RIIIa as the gamma-chain dimer was confirmed by elution of the protein from the anti-Fc gamma RIII immunoabsorbent with 1% Nonidet P-40 detergent and reimmunoprecipitation with anti-gamma-chain antibody. Phosphoamino acid analysis showed that the gamma-chain exclusively contained phosphotyrosine. The gamma-chain was also phosphorylated when electropermeabilized cells were activated by cross-linking Fc gamma RIIIa. The gamma-chain may play an important role in signal transduction via Fc gamma RIIIa in human macrophages. PMID- 8245473 TI - Macrophage progenitors from mouse bone marrow and spleen differ in their expression of the Ly-6C differentiation antigen. AB - Using mAb and magnetic and fluorescence-activated cell sorting, together with culturing in semisolid culture medium, we show that essentially all splenic macrophage progenitors are Ly-6ChiMac-1-. By contrast, bone marrow macrophage progenitors were more heterogeneous in Ly-6C expression, the majority being restricted to the Ly-6CdimMac-1- and the Ly-6C-Mac-1- subsets. These findings provide immunophenotype evidence that macrophages in the spleen and bone marrow arise from different progenitors. PMID- 8245474 TI - Induction by chemokines of lipid mediator synthesis in granulocyte-macrophage colony-stimulating factor-treated human neutrophils. AB - In recent years, there has been a growing body of evidence suggesting that IL-8 and granulocyte-macrophage CSF (GM-CSF) play an important role in inflammatory processes. We show that after GM-CSF treatment, the exposure of human neutrophils to IL-8 results in the synthesis of leukotriene (LT)B4 and platelet-activating factor. In GM-CSF-treated cells, IL-8 induced a concentration-dependent synthesis of both lipid mediators, with a threshold at 10 to 30 nM, suggesting that IL-8 could stimulate phospholipase A2 activity, an enzyme essential for both syntheses. Accordingly, IL-8 induced a substantial release of 3H-arachidonic acid in GM-CSF-treated PMN. It was also found that IL-8 activates the neutrophil 5 lipoxygenase (5-LO), the other key enzyme in LT biosynthesis. IL-8 induced 5-LO activation in a time- and concentration-dependent manner, with a threshold at 1 nM, and prior treatment of neutrophils with GM-CSF enhanced this effect of IL-8 over the 1 to 300 nM range. Neutrophil-activating peptide-2 and the melanoma growth-stimulatory activity, two peptides that are closely related to IL-8, also had the ability to activate the 5-LO and stimulate LT synthesis, albeit less potently than IL-8. Finally, pertussis toxin and the 5-LO translocation inhibitor, MK-886, both blocked the IL-8-elicited 5-LO activation. Taken together, our results raise the possibility that the combined presence of IL-8 and of GM-CSF at inflammatory foci could result in the synthesis of platelet activating factor and LTB4 by neutrophils, thereby contributing to the amplification of the inflammatory response. PMID- 8245475 TI - Multiple sites on IL-8 responsible for binding to alpha and beta IL-8 receptors. AB - To define the structural features important for IL-8 binding to its two known receptors, mutants of IL-8 and melanoma growth-stimulating activity (MGSA) and chimerae consisting of segments of these two chemokines were constructed and purified from the pGEX 2T Escherichia coli expression vector. IL-8 alpha and beta receptors were expressed stably and individually in 293 kidney epithelial cells and HL60 human leukemia cells. The Kd for IL-8 itself and copy numbers for both receptors in transfected cells were comparable. Competition binding with 125I labeled IL-8, however, showed large differences for several of the IL-8 mutants between alpha and beta receptors. The amino-terminal ELR sequence was important for IL-8 binding to the alpha receptor, but not sufficient for high affinity binding. Both rabbit IL-8 and MGSA share the ELR sequence with human IL-8, but compete poorly with it. The carboxyl terminus distal to amino acid 50 does not seem to mediate high affinity binding to the alpha receptor. A rabbit IL-8/human IL-8 chimera that differs in only eight amino acids from the human IL-8 sequence, was 150-fold lower in its affinity for the alpha receptor than human IL-8. In contrast, both the amino and carboxyl termini appear to be important for binding to the beta receptor. If the ELR sequence of IL-8 was substituted with alanines or if the carboxyl terminus distal to C50 was replaced with the MGSA sequence, a reduction occurred in binding competition. If both changes were introduced simultaneously, binding was abolished. Binding of MGSA was completely prevented by replacement of the ELR sequence with alanines. Ca2+ mobilization in HL60 cells transfected with the alpha or beta receptor was used to assess cell stimulation. The various mutant forms of IL-8 induced receptor activity with a pattern of sensitivity parallel to the competition binding affinities, indicating that both receptors are active. PMID- 8245476 TI - Binding of monomeric human IgG defines an expression polymorphism of Fc gamma RIII on large granular lymphocyte/natural killer cells. AB - Human PBL express one or more of the three classes of Fc receptors for IgG (Fc gamma receptors, I, II, and III). Each type of Fc gamma receptor has a characteristic binding pattern for isotypes of human and mouse IgG. Large granular lymphocyte/NK cells (LGL/NK cells) express the transmembrane form of Fc gamma RIII (Fc gamma RIIIa) on their surface, whereas polymorphonuclear neutrophils (PMN) express the glycosyl phosphatidyl inositol-linked receptor that is the product of the Fc gamma RIIIB gene. Fc gamma RIII has been reported to have low affinity for monomeric IgG, regardless of the cell type on which it is expressed. This study demonstrates specific and saturable binding of monomeric human IgG (M-IgG) to Fc gamma RIIIa on LGL/NK cells but not to Fc gamma RIIIb on PMN. M-IgG binding revealed an expression polymorphism between individuals, those with either high (12,000 to 15,000) or low (4,000 to 7,000) binding sites per cell. Both high and low M-IgG-binding LGL/NK cells bound hulgG1 and IgG3 but did not bind IgG2 or IgG4. The high/low Fc gamma RIIIA expression polymorphism correlated with the ability of an individual's LGL/NK cells to perform antibody dependent cellular cytotoxicity of erythroid cell targets. PMID- 8245477 TI - Increased gene expression of acetylcholine receptor and myogenic factors in passively transferred experimental autoimmune myasthenia gravis. AB - The passive transfer of myasthenia gravis by injection of mAb against muscle acetylcholine receptor (AChR) alpha-subunit, results in increased expression of AChR subunit genes, mainly at synaptic regions. The gene expression of AChR and of other muscle-specific proteins is regulated in a similar manner in passively transferred experimental autoimmune myasthenia gravis (EAMG) and in AChR-induced EAMG. Administration of AChR-specific mAb leads to a significant reduction in muscle AChR content and to an elevation in the mRNA levels corresponding to the adult, synaptic type of the receptor, as shown by Northern blot and in situ hybridization analyses. The mRNA levels of the myogenic factors myogenin and MRF4 are also increased moderately, whereas MyoD transcript levels remain unchanged. Thus, passive transfer of EAMG by mAb directed to defined epitopes of AChR alpha subunit provides a suitable model for analyzing and following the cascade of molecular events triggered by anti-AChR immunopathologic antibodies and may shed light on the regulatory mechanisms underlying the human disease as well. PMID- 8245478 TI - Interferon-gamma prevents graft-versus-host disease after allogeneic bone marrow transplantation in mice. AB - Lethally irradiated C3H/Law mice were injected (i.v.) with C57BL/Rij allogeneic bone marrow cells to induce a delayed type graft-vs-host disease (GVHD). Signs of GVHD first became apparent in the third week after transplantation. The disease resulted in a mortality rate of 70% at 80 days. Treatment with IFN-gamma twice weekly, for a period of 6 wk, starting at the time of bone marrow transplantation (BMT), completely prevented overt GVHD, as evidenced by a lack of diarrhea and no mortality during the follow-up period of 100 days after BMT. Also, the histologic GVHD lesions in the gastrointestinal tract were almost completely abrogated by the IFN-gamma treatment. All long term survivors were proven to be chimeras. During the induction phase of GVHD, the number of Con A-induced, IFN-gamma producing cells in the spleen was significantly reduced in the IFN-gamma-treated mice as compared with control mice. These results suggest that the normally enhanced production of endogenous IFN-gamma in the spleen at the time of hematopoietic reconstitution after BMT is down-regulated by exogenously administered IFN-gamma. This cytokine-mediated strategy to prevent GVHD might be an alternative to the current strategy of in vitro depletion of T cells for allogeneic BMT. PMID- 8245479 TI - Human T cell clones reactive against U-small nuclear ribonucleoprotein autoantigens from connective tissue disease patients and healthy individuals. AB - SLE and mixed connective tissue disease (MCTD) are characterized by the presence of high titers of autoantibodies against uridine-rich RNA-small nuclear ribonucleoprotein (snRNP) Ag. Because the presence of such snRNP-reactive autoantibodies has recently been shown to be associated with polymorphisms of HLA, this study was undertaken to determine whether snRNP-reactive T cells could be identified and characterized from patients. PBMC were stimulated with affinity purified snRNP Ag and cloned by limiting dilution in the presence of rIL-2 and rIL-4, snRNP-reactive human T cell clones were generated from three patients and two healthy blood donors who possessed disease-associated HLA genotypes. The cell surface phenotype of clones determined by flow cytometry was CD3+, CD4+, CD45RO+, TCR V alpha beta+. TCR V beta analysis, performed using V beta-specific primers and polymerase chain reaction, revealed that the T cell lines generated were clonal; a limited number of TCR V beta genes were expressed among the clones tested. All clones tested by mAb blocking of Ag-induced proliferation were restricted by HLA-DR. Several T cell clones were identified that were specific for B'/B or D polypeptides. These results demonstrate that snRNP-reactive T cells can be isolated from SLE and MCTD patients in vitro, and that Ag-driven expansion of such T cells could play a role in the immunopathogenesis of these diseases in vivo. PMID- 8245480 TI - C33 antigen and M38 antigen recognized by monoclonal antibodies inhibitory to syncytium formation by human T cell leukemia virus type 1 are both members of the transmembrane 4 superfamily and associate with each other and with CD4 or CD8 in T cells. AB - C33 Ag and M38 Ag had been identified by mAb inhibitory to HTLV-1-induced syncytium formation. The cDNA encoding C33 Ag had revealed that it belongs to the newly defined transmembrane 4 superfamily (TM4SF). M38 Ag was detected on virtually all human cell lines and fresh leukocytes except for most granulocytes. It was also expressed on a mouse hybrid cell clone containing human chromosome 11q23-pter. Immunoprecipitation and immunoblot analyses identified a monomeric 26 kDa protein. The M38 epitope was dependent on S-S bonding. These characteristics were very similar to those reported for TAPA-1 (the target of antiproliferative antibody-1), which also belongs to TM4SF as C33 Ag. We therefore cloned the cDNA of human TAPA-1 and expressed it in COS cells. M38 indeed reacted with COS cells expressing human TAPA-1. We concluded that M38 Ag was identical to TAPA-1. To further investigate the biologic functions of C33 Ag and M38 Ag (TAPA-1) and their roles in HTLV-1-induced syncytium formation, molecules associated with these Ag were examined in T cells. Immunoprecipitation from surface-iodinated cell lysates revealed that proteins co-precipitated by C33 and M38 were mostly common including each other. Sequential immunoprecipitation-immunoblot experiments confirmed that C33 Ag and M38 Ag (TAPA-1) were associated with each other. The association was further confirmed in BHK cells doubly transfected with human cDNA for C33 Ag and TAPA-1. We extended similar analyses and found that C33 Ag and M38 Ag (TAPA-1) were regularly associated with CD4 or CD8. The association of these Ag on the cell surface was further supported by co-modulation of M38 Ag (TAPA-1), CD4 and CD8 with C33 Ag. This is the first time that a physical association between the members of TM4SF is demonstrated. Furthermore, the regular association of C33 Ag and M38 Ag (TAPA-1) with CD4 or CD8 might indicate that they play a role in expression and/or function of the CD4/CD8 co-receptor complex. PMID- 8245481 TI - Analysis of glomeruli-eluted Gp330 autoantibodies and of Gp330 antigen of Heymann nephritis. AB - Quantity, charge, IgG isotype, Ag reactivity of glomerular gp330 autoantibodies (gp330Ab), and the charge of the putative Ag, gp330, were studied in active Heymann nephritis. Gp330 was anionic with an isoelectric point of 4.6. Despite variation in C3 glomerular immunofluorescence staining, C5b-9 staining was seen in all rats. Positive correlation was seen between glomerular gp330Ab and abnormal 24-h proteinuria (r = 0.637, p = 0.008), which appeared to require a certain threshold level of gp330Ab. Positive correlation was also seen between serum and glomerular gp330Ab at time of death (r = 0.55, p < 0.05). No differences were seen in charge, IgG isotypes of glomerular gp330Ab, or reactivity to gp330 by Western analysis in rats with or without abnormal proteinuria. Comparison of gp330Ab IgG isotypes in immune sera and glomeruli showed no difference in IgG1 and IgG2a, but IgG2b was significantly lower in glomeruli (p < 0.0001). CONCLUSIONS: 1) this is the first documentation that gp330 is highly anionic; 2) regardless of the degree of proteinuria, all immunized rats deposit in their glomeruli a qualitatively similar type of gp330Ab (charge, IgG isotype, complement activation, gp330 reactivity); 3) despite the highly anionic nature of gp330, anionic gp330Ab accumulate in glomeruli; and 4) complement activation appears to occur in every rat with gp330Ab in glomerulus and development of proteinuria is dependent on a certain threshold level of gp330Ab in the glomerulus which, in turn, correlates with the serum level of gp330Ab. PMID- 8245482 TI - Expression of alpha-3, alpha-5, and beta-4 neuronal acetylcholine receptor subunit transcripts in normal and myasthenia gravis thymus. Identification of thymocytes expressing the alpha-3 transcripts. AB - We used amplification of reverse transcribed RNA to investigate the thymic transcription of genes encoding for subunits of neuronal acetylcholine receptors (AcChR). All thymic tissues investigated, which include normal thymi, myasthenic hypertrophic thymi, and both myasthenic and nonmyasthenic thymomas, express transcripts encoding for the alpha-3, the alpha-5, and the beta-4 subunits of AcChR. The thymic transcription of at least three different genes encoding for neuronal AcChR subunits suggests there is expression of neuronal AcChR in thymus. Amplification of reverse transcribed RNA and Northern blotting of thymic and peripheral immunocytes showed that thymocytes but not peripheral lymphocytes express alpha-3 AcChR subunit transcripts. This finding suggests that within the T cell lineage there is transcriptional regulation of the alpha-3 AcChR gene. PMID- 8245483 TI - Constitutive expression of IL-6 receptors and their role in the excessive B cell function in patients with systemic lupus erythematosus. AB - Excessive B cell function including autoantibody production is a common feature of SLE and considered to be intimately associated with spontaneous lymphokine secretion by themselves. To clarify roles of IL-6/IL-6 receptor autocrine activation pathway in autoantibody production observed in patients with SLE, we studied expression and function of IL-6 receptors in comparison with those of IL 2 receptors, Tac on SLE B cells. IL-6 receptors and IL-2 receptors have been detected on B cells in the blood without any in vitro stimuli in most patients with SLE. The introduction of anti-IL-6 receptor antibody, which inhibits binding to the receptors of IL-6, and anti-IL-2 receptor antibody, anti-Tac to the cultures of SLE B cells resulted in potent inhibition of spontaneous production of polyclonal Ig and anti-DNA autoantibodies. In addition, fresh SLE B cells secreted high levels of IL-6 without any in vitro stimuli. These results indicate that constitutive expression of IL-6 receptors on B cells in conjunction with spontaneous IL-6 production by B cells induces autocrine B cell activation, which may lead to B cell hyperactivity and autoantibody secretion in SLE patients. Dysregulation of B cell activity observed in patients with SLE could thus be, at least in part, independent of T cell help. PMID- 8245484 TI - Role of gamma delta T cells in murine collagen-induced arthritis. AB - Murine collagen-induced arthritis (CIA) is a T cell-mediated disease which is induced by injection of type II collagen. Previous studies have shown that CD4+ cells which express particular V beta TCR genes are involved in the induction of arthritis in this model. In the present report we demonstrate that CD4-, CD8-, TCR gamma delta cells are present in arthritic joints, expanded in peripheral lymphoid tissue of DBA/1 lac J mice with CIA, and respond in vitro to the anti TCR gamma delta mAb UC7-13D5 (13D5). In order to directly investigate the role of the gamma delta TCR in murine CIA, DBA/1 lac J mice were injected with 13D5 before or 40 days after injection of type II collagen. Our results demonstrate that i.p. injections of 13D5 initiated 1 day before injection of type II collagen significantly delays both the onset and severity of CIA compared with treatment with type II collagen alone. In contrast, anti-TCR gamma delta mAb injection of arthritic mice 40 days after collagen injection resulted in the rapid onset of severe arthritis which was accompanied by increased bone erosion and cell infiltration into inflamed joints compared with arthritic mice injected with either control hamster IgG or F(ab')2 fragments of 13D5. Arthritic mice injected with intact 13D5 rapidly lost weight, suggesting that 13D5 may induce a cytokine mediated syndrome similar to that observed in mice and humans after the injection of anti-CD3. Flow cytometry analysis of joint cells isolated after collagenase digestion from arthritic mice demonstrated that 13D5 injection induces the accumulation of CD4-, CD8-, PgP-1 (CD44)+ cells within arthritic joints, whereas arthritic joints from mice injected with control hamster IgG contained cells with a CD4+, CD8- phenotype. CD3+ T cell lines which express the gamma delta TCR from inflamed joints of arthritic mice were established and examined for V gamma usage by the polymerase chain reaction. V gamma 2 rearrangements were predominant in both T cell lines established from inflamed synovium as well as freshly isolated synovial cells from arthritic mice, whereas synovial cells from nonarthritic mice did not demonstrate V gamma 2 rearrangements. Taken together, the results described in this report suggest a direct role for gamma delta TCR T cells in the pathogenesis of CIA in DBA/1 lac J mice. PMID- 8245485 TI - Preferential use of a H chain V region in antitumor-associated glycoprotein-72 monoclonal antibodies. AB - The DNA sequence of the mouse H chain V regions from five hybridomas directed against the human tumor Ag tumor-associated glycoprotein-72 (TAG-72) have been determined. This includes a previously determined VH gene sequence from a first generation anti-TAG-72 mAb, B72.3, and the VH gene sequences from four second generation anti-TAG-72 mAb, CC49, CC83, CC46, and CC92. A sequence comparison revealed a high degree of shared sequence identity between the five productively rearranged VH genes, suggesting derivation from a common germ line V region gene. In the process of cloning the unrearranged germ line gene, two highly related VH germ line genes were identified and designated VH alpha TAG-1 and VH alpha TAG-2. A comparison of the productively rearranged anti-TAG-72 VH sequences with the two germ line VH genes demonstrated that they were all derived from VH alpha TAG-1. In contrast, the L chain V regions are all derived from separate germ line V region genes. The preferential use of VH alpha TAG-1 in these five mouse hybridomas suggests that VH alpha TAG-1 is a preferred anti-TAG-72 H V chain region germ line gene and that the H chain plays a predominant role in the recognition of this Ag. PMID- 8245486 TI - Further characterization of the interaction between the C1q subcomponent of human C1 and the transmembrane envelope glycoprotein gp41 of HIV-1. AB - Previous studies have provided evidence for activation of the human C1 complex by HIV-1, resulting from direct interaction between C1q and the external portion of the viral transmembrane envelope protein, rsgp41. The present study was undertaken to locate more precisely, within C1q and rsgp41, the sites involved in the C1/HIV-1 interaction. Using a solid phase binding assay, we showed that 125I labeled C1q binding to rsgp41 was dose dependent, saturable, and comparable with binding of C1q to IgG-OVA immune complexes. The globular and, to a lesser extent, the collagen-like regions of C1q both bound to rsgp41. In contrast, the globular region of C1q inhibited the C1q/rsgp41 interaction, whereas the collagen-like region of C1q did not. A series of peptides covering the putative C1q-binding site on gp41 (positions 590-613 of gp160) were synthesized and used as competitors in the C1q-rsgp41-binding assay. Peptide 601-613 (GIWGCSGKLICT) inhibited C1q binding the most efficiently, with 50% inhibition at a concentration of 100 microM. This peptide also inhibited binding of C1q to rsgp36, the protein of HIV-2 homologous to rsgp41. The inhibitory effect of this peptide was dependent in part on the presence of the S-S bridge normally connecting Cys 605 to Cys 611 because reduction of this bond significantly reduced its efficiency. These data suggest that the C1q/HIV-1 interaction involves a site on C1q located within the globular regions, and a major site located within the immunodominant domain of HIV-1, which shares homology with the corresponding region of HIV-2. PMID- 8245487 TI - TSG-6: a TNF-, IL-1-, and LPS-inducible secreted glycoprotein associated with arthritis. AB - TSG-6 (TNF-stimulated gene 6) was originally discovered by differential screening of a cDNA library prepared from TNF-stimulated human diploid FS-4 fibroblasts. We show that the 35-kDa protein encoded by TSG-6 was undetectable in the medium of untreated FS-4 cultures, whereas its production reached approximately 1400 and 700 ng/10(6) cells after 24-h treatment with IL-1 or TNF, respectively. Stimulation of TSG-6 protein and mRNA levels was also demonstrated in normal human mononuclear cells by treatment with TNF and, especially, by LPS. In view of the inducibility of TSG-6 by inflammatory cytokines and its earlier demonstrated affinity for hyaluronan, we examined the presence of TSG-6 protein in the synovial fluids from patients with various forms of arthritis. TSG-6 protein was undetectable in the joint fluids of persons with no known history of arthritis, but high levels of TSG-6 oere demonstrated in the synovial fluids of a majority of arthritis patients. TSG-6 protein was also detected in the sera of some of the arthritis patients, albeit at concentrations that were less than in the joint fluids. To investigate the source of TSG-6 in the synovial fluids, we examined the production of TSG-6 protein in cultures of synovial cells. Synoviocytes from rheumatoid arthritis patients produced TSG-6 protein constitutively, and this production was increased by treatment with TNF or IL-1, but not with TGF-beta. Steady-state levels of TSG-6 mRNA were also increased in synoviocytes after treatment with TNF or IL-1. The presence of high levels of TSG-6 protein in the synovial fluids of arthritis patients and its inducibility by inflammatory cytokines in fibroblasts, mononuclear cells, synoviocytes, and chondrocytes suggest a role for TSG-6 in arthritis and inflammation. PMID- 8245488 TI - Influence of a recombinant human soluble tumor necrosis factor receptor FC fusion protein on type II collagen-induced arthritis in mice. AB - A recombinant human TNF receptor Fc fusion protein (rhuTNFR:Fc) was assessed for antiarthritic activity using murine type II collagen-induced arthritis in mice. DBA/1 mice were immunized with bovine type II collagen and treated with rhuTNFR:Fc either from day 21 to day 28 (preventative protocol), or after disease onset for fourteen days (therapeutic protocol). Control mice received either sterile saline or human serum albumin injections. rhuT-NFR:Fc treatment significantly reduced both the incidence and the severity of collagen-induced arthritis in the preventative protocol. Mice receiving rhuTNFR:Fc therapeutically progressed to less severe disease than did control animals, and the arthritis index in rhuTNFR:Fc treated mice was significantly lower than the index in control mice from 7.5 weeks after treatment. The antibody response to collagen was significantly reduced by treatment with rhuTNFR:Fc in both the preventative and therapeutic protocols. No difference was observed in the proliferative response to type II collagen or Con A, but the response to LPS was significantly lower in rhuTNFR:Fc treated mice at the conclusion of both the preventative and therapeutic trials. The results suggest that rhuTNFR:Fc may have both immunosuppressive and antiarthritic properties in this experimental model, and may represent a useful approach to the treatment of autoimmune arthritis. PMID- 8245489 TI - Implantation of dopaminergic cells in the treatment of parkinsonism. PMID- 8245490 TI - IMC (Amendment) Ordinance 1992. PMID- 8245491 TI - Perinatal mortality in caesarean section: a disturbing picture of unfulfilled expectations. AB - Indications for caesarean section had been studied in a 2-year period and the incidences were compared to that of the same 15 years back. Though there are more incidences of caesarean section, still perinatal death is a major concern to all. The study included a total of 291 perinatal deaths of which there were 208 early neonatal deaths and 83 stillbirths over a period of 2 years from January, 1990 to December, 1991. Caesarean section is being increasingly performed for foetal interest, but this study reveals that perinatal mortality is still high though cesarean section rate has increased in recent times. PMID- 8245492 TI - Effect of pH adjustment of bupivacaine on onset and duration of epidural anaesthesia. AB - The effect of pH adjustment of bupivacaine on onset and duration of sensory and motor block was studied in 45 patients undergoing lower abdominal and lower limb surgery. The study was done in 3 groups with the mean pH adjustment of 5.85, 7.14 and 7.21. Each patient was given 30 ml of 0.4% pH adjusted bupivacaine solution in epidural space. Onset of sensory block and duration were noticed by pin-prick method while motor block was observed by asking the patient to raise the thigh. It was observed that increase in pH of bupivacaine solution quickens the onset of sensory and motor block and increases the overall duration of neural blockade. PMID- 8245493 TI - Burns in elderly patients. AB - Admissions in the burn unit of IPGME & R, Calcutta over a period of 5 years from 1985 to 1990 have been reviewed. There were 1980 patients who had been treated and 84 of them were of 60 years of age or above. Mortality rate in elderly patients with burns had been found to be much higher. High mortality rate might be due to pre-existing medical problems and low general condition due to aging. Most of them were treated with conservative surgery. Early excision and grafting was the surgical approach in few cases with discouraging results. PMID- 8245494 TI - Ischaemic enteritis and colitis. PMID- 8245495 TI - Sporotrichosis. PMID- 8245496 TI - Haemangiopericytoma of urinary bladder. PMID- 8245497 TI - Meige's disease. PMID- 8245498 TI - Various questions on AIDS. PMID- 8245499 TI - Medical practitioners and 'golden handshake' in industry. PMID- 8245500 TI - Medical education. PMID- 8245501 TI - Modern prophylactic treatment of animal bite cases. PMID- 8245502 TI - Cyclosporin A rapidly inhibits epidermal cytokine expression in psoriasis lesions, but not in cytokine-stimulated cultured keratinocytes. AB - To better understand the cellular target(s) of cyclosporin action in psoriasis, we have studied the effects of systemic short-term (7 d), low-dose (3-7.5 mg/kg) cyclosporin A administration on the expression of the cytokines interleukin (IL) 8 and IL-1 beta in psoriatic lesions. RNA blot hybridization analysis of pretreatment keratome biopsies revealed that expression of both cytokine mRNAs was highly variable from patient to patient. Significant covariation of both cytokine mRNA levels was noted (r = 0.86, p < 0.0001). However, there was no significant correlation between expression of either cytokine and clinical severity, as measured by the pretreatment Psoriasis Area and Severity Index (PASI). IL-1 beta protein levels measured by enzyme-linked immunosorbent assay (ELISA) were highly correlated with IL-1 beta mRNA levels, indicating that the differences in transcript levels accurately reflect differences in epidermal cytokine protein. Significant reductions in both cytokine transcripts and in IL-1 beta immunoreactive protein were noted in the high expression subgroup after 1 week of cyclosporin A therapy, prior to detectable clinical improvement. In contrast to its pronounced effects on epidermal cytokine expression in vivo and the allogeneic mixed lymphocyte reaction in vitro, cyclosporine A did not inhibit the induction of intercellular adhesion molecule (ICAM)-1 or IL-8 mRNAs by cultured keratinocytes in response to IL-1 beta or the combination of tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma. These data suggest that epidermal keratinocytes respond to signals produced by activated T cells by coordinate expression of multiple cytokines, and that cyclosporin A acts primarily through blockade of T cells, rather than through keratinocyte activation. PMID- 8245503 TI - GRO-alpha mRNA is selectively overexpressed in psoriatic epidermis and is reduced by cyclosporin A in vivo, but not in cultured keratinocytes. AB - Interleukin (IL)-8 and gro peptides are members of the intercrine-alpha family of chemotaxins known to be present in biologically active form in psoriasis lesions. However, the relative contribution of the three different gro genes to the expression of this material is unknown, as is the stimulus for gro overexpression in psoriatic lesions. To address these questions, Northern blot and semiquantitative polymerase chain reaction analysis were performed on RNA extracted from keratome biopsies of normal skin, untreated plaques of psoriasis, or plaques treated for 1 week with low-dose cyclosporin A (CsA). Northern blot analysis revealed a significant correlation between gro and IL-8 mRNA levels in psoriasis lesions from 26 different individuals (r = 0.61, p = 0.0009), and overexpression of gro was markedly reduced by CsA prior to detectable clinical improvement (79.3%, p = 0.01, n = 22). To determine which form(s) of gro were overexpressed in psoriatic lesions, total keratome RNA (1 microgram) was analyzed by semiquantitative reverse transcription-polymerase chain reaction (SQRT-PCR). In five patients known to markedly overexpress gro and IL-8 mRNAs by Northern blotting, gro-alpha was approximately six times more abundant than gro-beta, and 25 times more abundant than gro-gamma. In cultured human keratinocytes, all three forms of gro mRNA were increased by IL-1 alpha or by interferon (IFN)-gamma plus tumor necrosis factor (TNF)-alpha. However, in contrast to the situation in vivo, CsA had no inhibitory effect on cytokine-stimulated gro expression in cultured keratinocytes. Taken together, these results demonstrate that the gro-alpha gene is selectively overexpressed in psoriatic lesions and strongly suggest that overexpression of gro is a keratinocyte response to activated T cells in psoriasis. PMID- 8245504 TI - Expression of gap junction proteins connexin 26 and 43 is modulated during differentiation of keratinocytes in newborn mouse epidermis. AB - We examined the expression of the gap junction proteins connexin 26 (Cx26), 32 (Cx32), and 43 (Cx43) in keratinocytes of newborn mouse epidermis to elucidate which connexins are expressed in keratinocytes in intact skin of newborn mice, and whether the expression of connexins is modulated during terminal differentiation of keratinocytes. Immunofluorescent staining using antibodies against Cx26, Cx32, and Cx43 combined with type-specific anti-keratin immunohistochemistry showed that Cx26 was expressed in keratinocytes in the granular layer and in the upper part of the squamous layer, whereas Cx43 was localized in keratinocytes in the basal layer and in the lower part of the squamous layer. No specific staining of Cx32 was found in mouse epidermis. Double staining of Cx26 and Cx43 revealed that some keratinocytes in the squamous layer expressed both connexins, but that in most cases localization of the two kinds of connexins was different, i.e., Cx26 was localized on the upper surface, whereas Cx43 was present on the lower surface of the plasma membrane of keratinocytes. Northern and Western blot analyses confirmed that Cx26 and Cx43, but not Cx32, were expressed at mRNA and protein levels in newborn mouse skin. These results suggest that the modulation of connexin expression from Cx43 to Cx26 takes place during terminal differentiation of keratinocytes in mouse epidermis. PMID- 8245505 TI - Localization of adducin in epidermis. AB - On the cytoplasmic side of the plasma membrane of erythrocytes there is a dense protein filament matrix that maintains the shape of the cells. The main constituents of this system, actin and spectrin, which have also been detected in keratinocytes and fibroblasts, are known to be linked in erythrocytes in a network structure by additional proteins such as band 4.1 and adducin. The interaction between actin and spectrin, mediated by adducin, is regulated by calmodulin and protein kinase C. Because we have previously found adducin in cultured keratinocytes, we investigated epidermis by immunochemical techniques. We found adducin to be localized at cell-cell contact sites in epidermis using affinity-purified antibodies against human erythrocyte adducin. Immunofluorescence of epidermis revealed an intense fluorescence in the basal layer, whereas stratum spinosum and stratum granulosum showed moderate staining. There was intense staining at sites of cell-cell contact in cultured human keratinocytes. Immunoblot analysis indicated the presence of adducin polypeptides of 103 kd and 97 kd in epidermis, but in cultured keratinocytes only the higher molecular weight form could be detected. This study indicates adducin, a regulatory protein in erythrocytes, is also present in epidermis. Its localization suggests that it may be involved in the formation of the microfilament matrix of the membrane skeleton at cell-cell contact sites. PMID- 8245506 TI - Talin: adherens junction protein is localized at the epidermal-dermal interface in skin. AB - The interaction between cells of the epidermis and the basal lamina is important for the integrity of the skin. Several hereditary and acquired diseases show changes at the dermal-epidermal interface due to loss of adhesion between basal cells and the basement membrane. The structures mediating this interaction are hemidesmosomes, which have been extensively characterized by biochemical, molecular biologic, and morphologic techniques. Recently, however, a group of adhesion molecules that are distinct from hemidesmosomes and that mediate cell matrix interactions was described in cultured fibroblasts, keratinocytes, and skin. These adhesion molecules, beta 1 integrins, have been shown to be present in the focal adhesion, a cell-matrix contact associated with microfilaments rather than intermediate filaments characteristic of hemidesmosomes. In cultured cells, integrins of the beta 1 family have been shown to be linked by a protein complex to actin filaments. In this study we describe the localization of talin, the binding protein for beta 1 integrins, and vinculin at the dermal-epidermal interface in skin with immunofluorescence and immunoblotting techniques. These data suggest the presence of a link between the cytoplasmic actin filament system in basal keratinocytes and the extracellular matrix. PMID- 8245507 TI - Predominance of epidermal CD8+ T lymphocytes in bullous cutaneous reactions caused by beta-lactam antibiotics. AB - The phenotype and functional characteristics of skin-infiltrating lymphocytes in beta-lactam antibiotic-induced vesiculobullous exanthemas were studied in vivo and in vitro. Immunohistochemical analysis demonstrated that CD8+ T lymphocytes were the predominant epidermal T-cell subset in these reactions. Epidermal T lymphocytes were isolated and expanded for in vitro studies. Fluorescence activated cell sorter analysis showed the majority of epidermal T cells to be CD3+, T-cell receptor alpha/beta+, CD4-, CD8+, and HLA-DR+, which correlated with the predominance of epidermal CD8+ T lymphocytes found in situ. Three CD8+ epidermal T-cell clones derived from cutaneous lesions proliferated in response to penicillin-pulsed autologous antigen-presenting cells but not allogeneic antigen-presenting cells, indicating that those clones were antigen and major histocompatibility complex specific. All T-cell clones produced significant amounts of interleukin-2, interferon-gamma, and granulocyte-macrophage colony stimulating factor. Additionally, the T-cell clones displayed cytotoxicity against epidermal cells in lectin-mediated cytotoxicity and against B-cell lines in T-cell receptor-triggered cytotoxicity. These data demonstrate the presence of epidermal drug-specific CD8+ T cells in bullous drug reactions. Because these CD8+ T cells have a cytotoxic potential, they may contribute to the necrosis of keratinocytes associated with drug-induced blister formation. PMID- 8245508 TI - Expression of diazepam-binding inhibitor peptide in human skin: an immunohistochemical and ultrastructural study. AB - Human diazepam binding inhibitor (DBI) was originally isolated from the brain and subsequently found to be present in several peripheral tissues. The various physiologic effects recently attributed to DBI include acting as an endogenous ligand for the central and peripheral (mitochondrial) benzodiazepine receptors. The present work provides, for the first time, evidence of DBI immunoreactivity in skin. DBI immunoreactivity was found in the epidermis, in the eccrine sweat and in sebaceous glands. Ultrastructurally, DBI was distributed throughout the cytoplasm. Although the physiologic role of DBI in skin is unknown, our results indicate that DBI may serve as an endogenous ligand for mitochondrial benzodiazepine receptors. Its activity could be related to the regulation of lipid and cholesterol synthesis in keratinocytes and sebaceous glands and to the secretion of sweat in sweat glands. PMID- 8245509 TI - Agarose gel keratinocyte outgrowth system as a model of skin re-epithelization: requirement of endogenous acetylcholine for outgrowth initiation. AB - To better understand the mechanisms of skin re-epithelization, we developed a simple technique that assays the outgrowth of human keratinocytes. Second-passage foreskin keratinocytes were inoculated at high cell density into 3-mm wells cut from agarose gels in standard 6-well tissue culture dishes. The cells settled on the dish bottom and formed a confluent colony. The cells at the periphery of the colony flattened, spread their cytoplasm, and moved away over the dish surface under the agarose gel. The morphology of migrating keratinocytes was observed microscopically through the transparent agarose, and the migration distance was measured after the gels were removed and after cells were fixed and stained. To determine which cell activities were involved in the outgrowth, the effects of cholinergic compounds on keratinocyte outgrowth were compared with their effects on keratinocyte proliferation, cell-plastic attachment, and spreading measured in separate sets of experiments. Outgrowth was inhibited by the specific inhibitor of acetylcholine synthesis bromoacetylcholine (0.05 mM) and restored by 5 mM exogenous acetylcholine. The irreversible muscarinic antagonist propylbenzilylcholine mustard (0.05 mM) abolished the restorative effects of exogenous acetylcholine, and also inhibited outgrowth of intact keratinocytes. In keratinocyte cell cultures, bromoacetylcholine stopped cell division. Propylbenzilylcholine mustard increased cell number, but interfered with cell plastic attachment and spreading. This suggests that cell-matrix attachment, spreading, and locomotion of human keratinocytes, but not mitosis, mediate the earliest stages of skin re-epithelization, and that endogenous acetylcholine regulates these keratinocyte functions. Specifically, keratinocyte acetylcholine is required to initiate outgrowth. PMID- 8245510 TI - Direct comparison of a cultured composite skin substitute containing human keratinocytes and fibroblasts to an epidermal sheet graft containing human keratinocytes on athymic mice. AB - This study compares two techniques for making cultured skin substitutes: a composite graft made of human fibroblasts and keratinocytes on a collagen glycosaminoglycan membrane (CG) and a cultured epidermal cell sheet graft (CEG), without a dermal component. The "take" and quality of these cultured skin substitutes were evaluated by placing them on full-thickness, excised wounds of athymic mice. These cultured skin substitutes were placed onto 2-X-2-cm wounds created on athymic mice. Mice were sacrificed at days 10, 20, and 42 with histologic sections obtained for light, electron, immunofluorescent, and immunohistochemical microscopy. "Take" was determined separately by a direct immunofluorescent stain for human leukocyte ABC antigens. There were ten mice of each graft type with at least two animals sacrificed at each time point. Results showed positive "take" for all animals. Grossly, there was little difference between the two graft types, with the CEG having occasional blister formation. By light microscopy, the CEG had a dissociation of dermis from epidermis until day 42, which was never apparent with the CG. By day 42, the CG had increased dermoepidermal interdigitations similar to rete ridges, with a mature epithelium. Neither of these findings were seen with the CEG. Immunofluorescent and immunohistochemical microscopy for type IV collagen and laminin, as well as electron microscopy, showed similar retardation of basement membrane formation with the CEG. Using this model, the composite graft had significant advantages over the epidermal sheet graft in the closure of full-thickness wounds. PMID- 8245511 TI - C3d,g deposits in inflammatory skin diseases: use of psoriatic skin as a model of cutaneous inflammation. AB - Recent studies in our laboratories have shown that human keratinocytes synthesize and secrete complement components including C3. Moreover, human keratinocyte derived C3 is regarded as a potential source of C3d,g, a recently described constituent of the sublamina densa region of normal epidermal basement membrane. Additionally, human keratinocyte-derived C3 may also contribute to epidermal basement membrane deposits of C3 in autoimmune or inflammatory skin disorders. To further our understanding of the specificity and origin of epidermal basement membrane C3 deposits in normal and diseased skin, we have characterized in situ deposits of C3 and C3 cleavage fragments in various inflammatory skin diseases and utilized a skin equivalent model to assess the deposition of C3 cleavage fragments in neo-basement membrane of epidermal outgrowths from normal or diseased human skin. C3d,g reactivity was found to be greater in all samples of inflamed skin, and typically associated with C3c reactivity at these sites. No immunoglobulins or other complement components were detected. When lesional psoriatic skin rich in epidermal basement membrane C3c was used in our organ culture system, C3 incorporation within neo-basement membrane was observed. These results show that human keratinocyte-derived C3 may contribute to inflammatory reactions in skin as well as account for deposits of C3d,g in normal epidermal basement membrane. PMID- 8245512 TI - Retinoid toxicity for fibroblasts and epithelial cells is separable from growth promoting activity. AB - Three different retinoids with widely varying capacity to stimulate skin repair in vivo and stimulate fibroblast and epithelial cell growth in vitro were examined for capacity to lyse the same cells at high concentrations. These included all-trans retinoic acid (RA), tetrahydro tetramethyl napthalenyl benzoic acid (TTNPB), and its biologically inactive structural analogue, meta-carboxy TTNPB. Despite their differing capacities to stimulate skin repair and cell growth, all of the agents were cytotoxic for fibroblasts and epithelial cells over the same range of concentrations (0.6-3 x 10(-5) M). Cytotoxicity for both fibroblasts and epithelial cells was blocked by addition of phosphatidylcholine to the cells along with the retinoid. In the presence of high concentrations of RA (0.75-3 x 10(-5) M) and phosphatidylcholine, proliferation was observed. The proliferative response was greater under these conditions than in the presence of an optimal concentration of RA (0.75-3 x 10(-6) M) without phosphatidylcholine. These data suggest that toxicity of retinoids can be separated, at least partially, from their growth-promoting activities. PMID- 8245513 TI - Podofilox-induced regression of Shope papillomas may be independent of host immunity. AB - We tested the hypothesis that infiltrating leukocytes might contribute to papilloma destruction following podofilox treatment. New Zealand White (NZW) rabbits were inoculated with cottontail rabbit papillomavirus (CRPV) onto abraded areas of the dorsal skin. At 21 d after viral inoculation, 5.0% podofilox solution was applied to some papillomas, whereas others were used as controls. Three rabbits were sacrificed at each of three different periods after treatment initiation (1, 4, and 7 d). Four monoclonal antibodies (MoAbs), RG-16 (for B cells), L11/135 (specific for T cells), 2C4 (specific for class II antigen), and Ki67 (specific for proliferating cells), were used in an immunohistochemical study. All positive cells and total cells in the field were counted with an ocular grid. After 1 d of treatment, proliferation of papilloma cells was strongly suppressed in treated papillomas, but leukocytic infiltration was not altered. At 4 d and 7 d of treatment, there were substantial increases (about two to three times) in the numbers of B and T cells and class II-expressing leukocytes. The upper layers of the papillomas were highly necrotic and cell proliferation was absent in all layers. These data support the view that podofilox has a direct toxic effect on papilloma tissue. Leukocyte infiltration is not strongly associated with papilloma tissue and may not contribute to papilloma destruction. PMID- 8245514 TI - Differentiation-associated localization of nPKC eta, a Ca(++)-independent protein kinase C, in normal human skin and skin diseases. AB - The expression of nPKC eta, a Ca(++)-independent isoform of protein kinase C in normal human skin, and skin from patients with psoriasis, squamous cell carcinoma, basal cell epithelioma, nevus pigmentosus, and seborrheic keratosis, were examined by immunohistochemical staining using a polyclonal antibody raised against a synthetic peptide at a diverse region of the nPKC eta molecule. In normal epidermis, the strongest staining was observed in the uppermost granular layer with no staining of the spinous or basal layers. The inner layer of the intra-epidermal eccrine duct was also strongly stained. Weak staining was observed in several layers of the outer root sheath of the follicular infundibulum. No staining was detected in the inner root sheath of the hair follicles, hair matrix, sebaceous gland, eccrine gland, intradermal eccrine duct, arrectores pilorum, melanocytes, Langerhans cells, fibroblasts, or blood vessels. In psoriatic skin, stained keratinocytes were distributed in the suprabasal layers with the most being observed in the uppermost layer and the least in layers closed to the basal layer. In squamous cell carcinoma, weak staining was observed in the keratotic cells around horny pearls. In the basal cell epithelioma and nevus pigmentosus, the cells were not stained, whereas in seborrheic keratosis, cells that stained were located in the granular layer. We conclude from the evidence presented above that nPKC eta is expressed in close association with epidermal differentiation in normal skin and skin diseases. PMID- 8245515 TI - Analyses of mixed melanogenesis in tyrosinase cDNA-transfected human amelanotic melanoma cells. AB - In the pigment cell the synthesis of tyrosinase and the formation of premelanosomes are independent, yet coordinated, processes. However, the interrelationship between the two processes has not been elucidated previously. In this study, an expression plasmid for human tyrosinase cDNA was constructed and transfected into a human amelanotic melanoma cell line, G-361. Stable transfected cells (G-CMHT-3) were obtained with high tyrosinase activity and distinct melanization occurred. As for the type of melanin, both pheo- and eumelanin contents increased in G-CMHT-3 cells. Interestingly, catalase activity as one of the other melanogenic enzymes was decreased in G-CMHT-3 cells. The decrease of catalase activity was considered to play a role in melanin-polymer formation, resulting in the increase of both pheo- and eumelanin contents. Under electron microscopic observation, dopa-oxidase-positive Golgi-associated endoplasmic reticulum of lysosome, coated vesicles, and premelanosomes were observed in pigmented G-CMHT-3 cells, and the expressed tyrosinase was considered to be well translocated to these organelles. In addition, the number of premelanosomes (stages I-III) as well as melanosomes (stage IV) increased in G CMHT-3 cells compared to those in G-361 cells. It is also noted that G-CMHT-3 cells showed more normal phenotype premelanosomes with occasional transitional premelanosomes exhibiting partial melanin polymer formation within their concentric whorl-like internal membranes. Furthermore, the number of eumelanosomes in G-CMHT-3 cells was much larger than that in G-361 cells. These results suggest that the tyrosinase introduced by its cDNA transfection induced active and structurally non-aberrant premelanosome formation resulting in the upregulated pheo- and eumelanogenesis. PMID- 8245516 TI - Phenotypic differences in cytokine responsiveness of hypertrophic scar versus normal dermal fibroblasts. AB - The alteration of normal dermal fibroblast function that leads to the development of hypertrophic scar after thermal injury is unknown. To determine functional differences that might explain this process, fibroblasts were cultured from biopsies of post-thermal injury mature hypertrophic scars and patient-matched normal skin. The mitogenic responses of scar cells to fetal bovine serum, epidermal growth factor (EGF), platelet-derived growth factor (PDGF), and tumor necrosis factor alpha (TNF alpha) were determined and compared to normal skin cells. Collagen synthetic rate was also compared in the presence and absence of transforming growth factor beta 1 (TGF beta 1). Whereas both scar and normal cells responded with increased thymidine uptake to serum and cytokines, the stimulation to EGF and serum was significantly lower in scar cells. In contrast, synthesis of collagen, but not of non-collagenous proteins, was increased in scar relative to normal cells, both basally and when stimulated with low doses of TGF beta 1. Additionally, the fraction of protein synthesized as collagen was significantly higher in scar fibroblasts. These results suggest that fibroblasts from hypertrophic scars demonstrate stable phenotypic differences in cytokine responsiveness in comparison to cells from unaffected skin. The increased rate of collagen synthesis and decreased responsiveness to mitogens are consistent with the increased extracellular matrix content and decreased cellularity of hypertrophic scars. PMID- 8245517 TI - Enhanced low-density lipoprotein degradation and cholesterol synthesis in monocyte-derived macrophages of patients with adult xanthogranulomatosis. AB - Adult xanthogranulomatosis is an uncommon disorder in which dermal macrophages accumulate cholesterol intracellularly despite normal plasma cholesterol levels. In an attempt to elucidate an underlying biochemical abnormality in this disorder, we studied the rates of 125I-labeled low-density lipoprotein degradation, and intracellular cholesterol synthesis, in human monocyte-derived macrophages of three patients with adult xanthogranulomatosis. In all three patients, the rates of cellular 125I-low-density lipoprotein degradation and of cholesterol synthesis were 22-37% and 14-84% higher than those of the respective normal controls (p < 0.01). These findings suggest that in MDM of adult xanthogranulomatosis patients, the uptake and degradation of low-density lipoprotein-derived cholesterol and intracellular cholesterol biosynthesis are enhanced. Because dermal macrophages are derived from blood monocytes, it is possible that such an enhancement might play a role in the accumulation of cholesteryl esters in the macrophages that form the xanthogranulomatosis lesions. PMID- 8245518 TI - Hematogenous spread of malignant melanoma cells in different stages of disease. AB - Patients with malignant melanoma and distant metastases generally have an unfavorable prognosis, with a median survival of about 6 months. The mechanisms of hematogenous spread and implantation of melanoma cells are, however, poorly understood, because the standard diagnostic methods are not sensitive enough to detect oligocellular micrometastases. Recently a method using reverse transcription and polymerase chain reaction to determine tyrosinase mRNA in peripheral blood, which indicates the presence of circulating melanoma cells, has been developed. We utilized this assay to examine blood samples of 56 patients with malignant melanoma in different stages of disease. In one of 10 patients in stage I (localized disease) and in six of 17 patients with regional lymph nodes metastases (stage II) tyrosinase mRNA was detected. Tyrosinase transcripts were found in all 29 patients with distant metastases (stage III). Interestingly, tyrosinase mRNA was also detected in six patients with metastatic amelanotic malignant melanoma. In contrast, tyrosinase mRNA was not detectable in any of 39 healthy subjects or 17 patients with other malignancies. These findings may be helpful to define a patient group at high risk for systemic spread of disease. PMID- 8245519 TI - Human oropharyngeal lesions with a defective Epstein-Barr virus that disrupts viral latency. AB - The Epstein-Barr virus (EBV), a human herpesvirus that is predominantly latent after infection, can be induced to replicate by deleted, rearranged EBV DNA from cultures of laboratory strain P3HR-1. Because mucosal surfaces are permissive of EBV replication, 101 oral biopsies from 70 Chinese and 5 American patients were examined for natural counterparts to tissue culture defective virus (WZhet), using as marker the abnormal juxtaposition of BamHI W and Z EBV DNA restriction fragments. Of the 49 oral biopsies that contained EBV DNA, 12 (24%) had the rearranged WZ fragment by polymerase chain reaction analysis: 3 (42%) of 7 EBV positive epithelial dysplasias or carcinomas, 6 (38%) of 16 hairy leukoplakias, and 3 (12%) of 25 nonmalignant salivary gland biopsies. Accompanying viral replication was confirmed by in situ cytohybridization and demonstration of the linear configuration of the genome in select WZhet-positive lesions. These findings indicate that defective EBV with the unusual property of disrupting EBV latency is prevalent in natural infections and may contribute to EBV's pathogenic diversity. PMID- 8245520 TI - The effect of decreasing amounts of live virus, while antigen content remains constant, on immunogenicity of Oka/Merck varicella vaccine. AB - The purpose of the study was to establish the minimum immunizing dose of stored Oka/Merck varicella vaccine and mimic effects of prolonged storage of vaccine at freezer temperatures. Ninety children were randomized to one of three groups: group A (1770 pfu/dose), group B (400-500 pfu/dose), or group C (80-160 pfu/dose). Seroconversion rates for groups A-C were 97%, 96%, and 92%, respectively. Group A and B vaccinees had higher antibody titers at 1 year than did group C subjects (P = .01). The difference between groups B and C approached significance (P = .058). At 1 year after immunization, 73%, 83%, and 80% in groups A-C, respectively, demonstrated cell-mediated responses. Since freshly reconstituted vaccine contains 1770 pfu/dose, vaccine failure from loss of potency is unlikely if vaccine is stored properly and administered promptly after reconstitution. PMID- 8245521 TI - Distribution of wild type 1 poliovirus genotypes in China. AB - Poliomyelitis remains an important public health problem in China. Most cases and outbreaks are associated with wild type 1 polioviruses. To obtain an overview of type 1 poliovirus transmission in China, partial genomic sequences were compared for 24 case isolates from 12 provinces. Because polioviruses evolve rapidly during infection of humans, the genetic relationships among isolates provide a measure of the extent of epidemiologic linkage among cases. The observed genetic relationships were complex: six different genotypes, apparently derived from five separate endemic origins, were found. One genotype was recombinant, having noncapsid sequences derived from the Sabin type 1 vaccine strain and capsid sequences derived from a genotype indigenous to several northern and eastern provinces. Some isolates from geographically separate provinces were closely related; other isolates were related to wild polioviruses found in neighboring countries. The combination of epidemiologic and virologic analyses may facilitate the development of more effective strategies for poliomyelitis eradication. PMID- 8245522 TI - Quinolinic acid in the cerebrospinal fluid of children with symptomatic human immunodeficiency virus type 1 disease: relationships to clinical status and therapeutic response. AB - Quinolinic acid (QUIN) is a neurotoxin implicated in the neurologic deficits associated with human immunodeficiency virus type 1 (HIV-1) infection. Forty children with symptomatic HIV-1 disease had elevated (P < .001) cerebrospinal fluid (CSF) QUIN levels (55.8 +/- 8.9 nM) compared with controls (14.9 +/- 3.0 nM). Age-adjusted CSF QUIN concentrations in HIV-1-infected children were predicted by the general index of mental abilities (GIMA, from an age-appropriate intelligence test; r = -0.45, P < .01). Zidovudine therapy reduced CSF QUIN from 64.1 +/- 16.3 to 19.7 +/- 5.2 nM (P < .01; N = 16) and increased GIMA from 76.8 +/- 5.2 to 87.2 +/- 6.3 (P < .001). Encephalopathic HIV-1-infected patients had higher CSF QUIN levels than patients without encephalopathy (79.6 +/- 16.1 vs. 32.7 +/- 6.7 nM, P < .01). CSF QUIN concentrations were also higher (P < .001) in patients who died < or = 3 years after their baseline assessment, compared with those who were still alive. These results warrant further investigation of CSF QUIN in HIV-infected children as a mediator of neurologic dysfunction and a supplemental marker of neurologic disease, particularly when combined with measures of neurocognitive functioning. PMID- 8245523 TI - Safety and immunogenicity of a fully glycosylated recombinant gp160 human immunodeficiency virus type 1 vaccine in subjects at low risk of infection. National Institute of Allergy and Infectious Diseases AIDS Vaccine Evaluation Group Network. AB - Recombinant gp160 derived from human immunodeficiency virus type 1 (HIV-1)IIIB and produced in mammalian tissue culture cells using a vaccinia virus expression system (rgp160-mam) was evaluated as a vaccine in combination with alum and deoxycholate adjuvant. Sixty low-risk, uninfected subjects received 12.5 micrograms, 50.0 micrograms, or adjuvant control at 0, 1, 6, and 12 months in a randomized, double-blind dose-escalation study. A single injection of 200 micrograms of vaccine was given at 18 months in an open study to 9 vaccines who had received 50 micrograms. The vaccine was safe. Six of 16 subjects receiving 50 micrograms developed neutralizing antibody to HIV-1IIIB. Seven of the 9 boosted with 200 micrograms of vaccine at 18 months developed neutralizing antibodies. Lymphocyte proliferation to rgp160-mam and baculovirus-derived rgp160 and rgp120 was induced in both groups (12.5 and 50.0 micrograms) and appeared after the first dose. Further studies with higher doses of rgp160-mam and vaccines derived from other strains of HIV-1 are warranted. PMID- 8245524 TI - Cross-reactive response to human immunodeficiency virus type 1 (HIV-1) gp120 and HLA class I heavy chains induced by receipt of HIV-1-derived envelope vaccines. AB - Autoantibodies specific to HLA class I antigens were detected in the sera of persons vaccinated with human immunodeficiency virus type 1-derived recombinant vaccines by using synthetic peptides representing the amino acid sequences recognized by an HLA class I/gp120 cross-reactive monoclonal antibody. Study subjects received recombinant vaccinia gp160 (vacc-env) alone, vacc-env followed by one dose of recombinant gp160 (rgp160, 640 micrograms), or four doses of rgp160 alone (640 or 80 micrograms). All sera from vacc-env/rgp160-vaccinated subjects contained HLA/gp120 cross-reactive antibodies, as did all sera from recipients of the rgp160 alone at 640 micrograms/dose. In contrast, none of the sera from subjects who received either the vacc-env alone or the 80 micrograms/dose rgp160 alone contained detectable HLA cross-reactive antibodies, and these same sera showed little or no envelope reactivity on Western blot. The results showed a striking correlation between immunogenicity and the induction of cross-reactive antibodies by the rgp160 vaccine. PMID- 8245525 TI - The association between circumcision status and human immunodeficiency virus infection among homosexual men. AB - To evaluate whether uncircumcised status is correlated with acquisition of human immunodeficiency virus (HIV), 502 homosexual men were surveyed; 85% were circumcised. HIV infection was significantly associated with uncircumcised status (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.2, 3.8), nonwhite race, intravenous drug use, sexual contact with an intravenous drug user, number of male partners, frequency of unprotected receptive anal intercourse, and with history of genital herpes, anal herpes, or syphilis. Uncircumcised status was significantly associated with older age, nonwhite race, and history of syphilis; it was inversely associated with intravenous drug use. Using logistic regression analysis, the adjusted OR for the association between HIV infection and uncircumcised status was 2.0 (95% CI, 1.0, 4.0). Uncircumcised homosexual men had 2-fold increased risk of HIV infection. The role of circumcision as an intervention strategy to reduce sexual transmission of HIV warrants consideration. PMID- 8245526 TI - Early abnormalities of the antibody response against Mycobacterium tuberculosis in human immunodeficiency virus infection. AB - Among human immunodeficiency virus (HIV)-infected persons, those who react against purified protein derivative (PPD) have higher risk of tuberculosis. Since PPD testing has limited predictive power in HIV-positive populations, new markers of antituberculous immunity were sought by analyzing antibodies to Mycobacterium tuberculosis antigens (PPD and its fraction A60) in 102 HIV-positive subjects, some PPD-positive and some PPD-negative, and in 23 HIV-positive tuberculosis patients. ELISA and Western blotting were used. Forty HIV-negative healthy subjects and 40 HIV-negative tuberculosis patients were evaluated as controls. While all those HIV-negative and PPD-positive had IgG antibodies recognizing the 38-, 28-, and 19-kDa M. tuberculosis antigens, only 26% of those HIV-positive and PPD-positive (all with < 400 CD4+ cells/mm3) and none of the HIV-positive tuberculosis patients recognized them, indicating that the lack of IgG against those antigens, in the presence of a specific IgM response, is a marker of immunodeficiency. PMID- 8245527 TI - Serum antibody opsonic activity against Actinobacillus actinomycetemcomitans in human periodontal diseases. AB - Actinobacillus actinomycetemcomitans is frequently associated with severe periodontitis. Many periodontitis patients have elevated levels of serum IgG antibodies to A. actinomycetemcomitans, but the role of these antibodies is unknown. This study evaluated the functional capacity of anti-A. actinomycetemcomitans IgG antibody to enhance phagocytosis of A. actinomycetemcomitans by polymorphonuclear leukocytes. Chemoluminescence assays were done using sera from 64 subjects, 61 of whom had severe periodontitis; results were compared with the subject's anti-A. actinomycetemcomitans IgG titer and avidity. There was a strong correlation between chemoluminescence and antibody log titer (P < .00001) and a weak correlation between chemoluminescence and antibody avidity (P < .05). The results support the hypothesis that anti-A. actinomycetemcomitans IgG antibodies are important in promoting phagocytosis and killing of A. actinomycetemcomitans. Subjects who develop high levels of highly avid antibodies against A. actinomycetemcomitans may have greater resistance to continued or repeated infection by this pathogen. PMID- 8245528 TI - Mucosal and systemic T helper cell function after intragastric colonization of adult mice with Candida albicans. AB - In 85% of adult DBA/2 mice inoculated intragastrically with Candida albicans, significant numbers of yeast cells were recovered from the gastrointestinal tract for up to 4 weeks, with the animals eventually clearing infection in the absence of systemic disease despite the occurrence of localized, self-limiting foci of mucosal involvement in their stomachs. Two major findings in colonized mice were defective production of IgA, interleukin (IL)-4, and IL-5 by Peyer's patches lymphocytes and increased numbers of interferon-gamma-producing T cells in mesenteric lymph nodes and spleens. Relatively low levels of circulating antibodies of T helper type 2 (Th2)-dependent isotypes were also found in colonized mice, which exhibited strong footpad responses and increased resistance to systemic reinfection. Unlike systemic challenge, gastrointestinal colonization of adult immunocompetent DBA/2 mice with C. albicans appears to be an effective stimulus for the systemic development of protective Th1 responses. PMID- 8245529 TI - Systemic cell-mediated immune reactivity in women with recurrent vulvovaginal candidiasis. AB - It has been suggested that impaired cell-mediated immunity (CMI) against Candida antigens is responsible for susceptibility to recurrent vulvovaginal candidiasis (RVVC) in adult women. To address this, we conducted a comprehensive longitudinal study examining in vivo and in vitro systemic CMI reactivity in RVVC patients. Results showed that RVVC patients frequently demonstrated a transient loss of Candida-specific delayed cutaneous skin test reactivity during episodes of symptomatic vaginitis. In contrast, in vitro peripheral blood lymphoproliferation and Th1-type lymphokine production by RVVC patients in response to a T cell mitogen and multiple Candida and bacterial antigens were similar to controls both during acute episodes of vaginitis and during periods of infection-free remission. These results suggest that women with RVVC have no detectable impairment of systemic CMI in peripheral blood and that transient reductions in skin test reactivity appear to be a result of vaginal Candida infection and not a predisposing factor to RVVC. PMID- 8245530 TI - Phagocytosis and stimulation of the respiratory burst in neutrophils by Pneumocystis carinii. AB - Phagocytosis and superoxide production of neutrophils stimulated with Pneumocystis carinii were studied using P. carinii obtained from rats and neutrophils and serum from healthy blood donors. Superoxide generation increased in a dose-related way to the number of pneumocysts added and was significantly higher when the parasites were opsonized. The use of heat-inactivated serum for opsonization reduced superoxide production somewhat, but not significantly. Preincubation of neutrophils with recombinant human granulocyte-macrophage colony stimulating factor (rHuGM-CSF) increased superoxide production in neutrophils stimulated with pneumocysts from 7.0 to 9.6 fmol/cell/20 min; however, the production in unstimulated cells increased by the same amount. Activation of the respiratory burst was paralleled by phagocytosis, and very few phagocytic vacuoles were found in neutrophils stimulated with unopsonized pneumocysts. By electron microscopy, various stages of cyst degradation were seen. PMID- 8245531 TI - Intracellular survival and multiplication of Toxoplasma gondii in astrocytes. AB - Primary neonatal murine astrocyte cultures were used to investigate the role of these glial cells in host defense of the central nervous system (CNS) against Toxoplasma gondii. For comparison, neonatal murine microglial cells were also studied. Microscopic analyses revealed that uptake of T. gondii into astrocytes was parasite-driven and was followed by uniform intracellular survival and multiplication of tachyzoites. Treatment of astrocytes with interferon (IFN) gamma and lipopolysaccharide (LPS) had no apparent effect on the survival or growth of T. gondii. Microglia, on the other hand, had both an intrinsic phagocytosis-associated antitoxoplasma activity and a nitric oxide-dependent inhibitory activity that was up-regulated by IFN-gamma and LPS. The results of this study suggest that in contrast to microglial cells, astrocytes may provide a safe harbor within the CNS for T. gondii. PMID- 8245532 TI - Strongyloides stercoralis: the first rodent model for uncomplicated and hyperinfective strongyloidiasis, the Mongolian gerbil (Meriones unguiculatus). AB - Strongyloidiasis is the most common endemic helminthiasis in several of the world's industrialized nations, yet relatively little is known about its basic biology and immunobiology because a practical rodent model for the investigation of this clinically important parasitism is lacking. This study reports such a model for use in the investigation of Strongyloides stercoralis infection. Normal male gerbils infected subcutaneously with 1000 infective filariform larvae harbored moderate numbers (83.6 +/- 27.6) of adult worms at 35 days after infection, and a low-grade infection persisted for at least 131 days mimicking the chronicity of human infections. Gerbils treated weekly with 2 mg of methylprednisolone acetate developed hyperinfective strongyloidiasis with up to 8000 autoinfective larvae occurring in these animals at postinfection day 21. Autoinfection never occurred in normal (untreated) gerbils. PMID- 8245533 TI - Global occurrence of Plasmodium vivax-like human malaria parasite. AB - A Plasmodium vivax-like human malaria parasite was recently identified from Madang, a holoendemic malarious region in Papua New Guinea. The complete nucleotide sequence of the circumsporozoite (CS) protein gene of this parasite is presented here. The CS protein of this parasite has an 11-mer repeat sequence and is different from the other known CS protein genes of human malaria parasites. However, it is identical to the CS protein gene of a monkey malaria parasite, Plasmodium simiovale. This P. vivax-like malaria parasite was found in Sepik, another malarious region of Papua New Guinea, and in Brazil, Indonesia, and Madagascar. No pure isolate of this parasite was identified. Specific oligonucleotide probes were used to determine relative proportion of the P. vivax like parasite in P. vivax (type 1 and type 2) mixed field isolates. Compared with P. vivax or Plasmodium falciparum, the circumsporozoite protein of P. vivax-like parasites showed markedly less polymorphism. PMID- 8245534 TI - Primary human immunodeficiency virus type 1 infection: review of pathogenesis and early treatment intervention in humans and animal retrovirus infections. AB - Primary human immunodeficiency virus type 1 (HIV-1) infection can present clinically as the abrupt onset of a febrile illness resembling acute mononucleosis. The symptoms coincide with high titers of culturable plasma viremia, cell-associated virus, and antigenemia, which rapidly decrease coincident with the emergence of detectable HIV-specific antibody and HIV specific cytotoxic T lymphocytes. This article reviews the human and animal model data on the virologic and immunologic events that occur during primary HIV-1 and animal retrovirus infections, evaluates the prophylactic treatment experience of retrovirus infections in the animal model, and provides a plausible rationale for treatment intervention of primary HIV-1 infection in humans. Recent work delineating the pathogenesis of primary HIV-1 infection provides insight into the major mechanisms of viral dissemination and host immune response. The results from retrovirus-infected animal models treated with antiviral agents suggests that therapy at the time of viral dissemination may be an effective strategy that may modify disease progression. Clinical trials to evaluate this approach are in progress. PMID- 8245535 TI - Quantitative cultures of the cell fraction and supernatant of bronchoalveolar lavage fluid for the diagnosis of cytomegalovirus pneumonitis in lung transplant recipients. AB - Results of quantitative shell vial cultures of cell and supernatant fractions of bronchoalveolar lavage (BAL) fluid from lung transplant recipients were compared with results of transbronchial lung biopsies. Cytomegalovirus (CMV) was present in both the cell and supernatant fractions, including 28 (80%) and 29 (83%), respectively, of 35 BAL samples with corresponding biopsies that showed evidence of CMV pneumonitis and 34 (15%) and 75 (33%), respectively, of 227 BAL samples with corresponding biopsies that did not. Cultures of unseparated BAL fluid had a similar yield to cultures of supernatant. Virus titers of cell fractions and supernatants from BAL samples with corresponding biopsies that were positive for CMV pneumonitis were higher than those from BALs with negative corresponding biopsies. Culture of the unseparated fluid is recommended for simplicity and to maximize detection of CMV. Quantitative results may help in the identification of CMV pneumonitis, but additional studies are needed before quantitative cultures can be recommended for routine use. PMID- 8245536 TI - Intravitreal ganciclovir concentration after intravenous administration in AIDS patients with cytomegalovirus retinitis: implications for therapy. AB - To determine whether therapeutic intravitreal concentrations of ganciclovir are achieved after intravenous administration, vitreous samples were obtained intraoperatively from 23 eyes of 22 AIDS patients with retinal detachments associated with cytomegalovirus (CMV) retinitis. The mean intravitreal ganciclovir concentration of all samples was 0.93 +/- 0.39 microgram/mL (3.6 +/- 1.5 microM). This level is near the published trough serum concentrations obtained with every-12-h intravenous dosing and well below the peak. It is significantly below the concentration of ganciclovir required to achieve 50% of viral plaque formation for many human CMV strains. Only a small decrease in vitreous drug levels was observed as a function of time after last dose. Intravenous administration of ganciclovir results in near-steady-state subtherapeutic intravitreal ganciclovir concentrations for many CMV isolates. This may explain the difficulty of long-term complete suppression of CMV retinitis. PMID- 8245537 TI - Direct detection of wild poliovirus circulation by stool surveys of healthy children and analysis of community wastewater. AB - Cartagena, Colombia, was one of the last cities in the Americas known to have endemic poliomyelitis. After 3 cases were identified in 1991, two approaches for detecting continued silent transmission of wild polioviruses within a high-risk community were used: stool surveys of healthy children and virologic analysis of community sewage. Wild type 1 polioviruses were isolated from 8% of the children studied and from 21% of sewage samples. The proportions of wild polioviruses, vaccine-related polioviruses, and nonpolio enteric viruses were similar for both approaches. Wild poliovirus sequences were also amplified directly from processed sewage samples by the polymerase chain reaction using primer pairs specific for the indigenous type 1 genotype. The last reported cases associated with wild polioviruses in the Americas occurred in Colombia (8 April 1991) and Peru (23 August 1991). Direct sampling for wild polioviruses in high-risk communities can provide further evidence that eradication of the indigenous wild polioviruses has been achieved in the Americas. PMID- 8245538 TI - Respiratory syncytial virus lung infection in infants: immunoregulatory role of infected alveolar macrophages. AB - Thirteen previously healthy children with acute onset of severe lower respiratory tract signs and symptoms underwent bronchoscopy and bronchoalveolar lavage (BAL) for diagnostic purposes. BAL samples were assessed for viral, bacterial, mycobacterial, and fungal cultures. Cytospin preparations of BAL cells were assessed for expression of respiratory syncytial virus (RSV), HLA-DR, interleukin 1 beta, and tumor necrosis factor proteins. Purified alveolar macrophages from 2 RSV-infected children were assessed for viral replication. Three children had bacterial pneumonia and 6 were infected with RSV. BAL cells from RSV-infected children demonstrated viral protein expression. Alveolar macrophages were the predominant cell type recovered by BAL and demonstrated coexpression of RSV, HLA DR, interleukin-1 beta, and tumor necrosis factor proteins. Purified alveolar macrophages from 2 RSV-infected children replicated RSV by infectious center assays. Thus, alveolar macrophages are infected by RSV in vivo and coexpress potent immunomodulatory molecules that potentially regulate the local immune response or lung injury due to this virus. PMID- 8245539 TI - Congenital yellow fever virus infection after immunization in pregnancy. AB - To determine whether yellow fever (YF) vaccine administered in pregnancy causes fetal infection, women who were vaccinated during unrecognized pregnancy in a mass campaign in Trinidad were studied retrospectively. Maternal and cord or infant blood were tested for IgM and neutralizing antibodies to YF and dengue viruses. One of 41 infants had IgM and elevated neutralizing antibodies to YF virus, indicating congenital infection. The infant, the first reported case of YF virus infection after immunization in pregnancy, was delivered after an uncomplicated full-term pregnancy and appeared normal. Congenital dengue 1 infection may have occurred in another case. The frequency of fetal infection and adverse events after such exposure could not be estimated; however, the neurotropism of YF virus for the developing nervous system and the now documented possibility of transplacental infection underscores the admonition that YF vaccination in pregnancy should be avoided. PMID- 8245540 TI - Conjunctival scarring in trachoma is associated with depressed cell-mediated immune responses to chlamydial antigens. AB - In vitro responses of peripheral blood mononuclear cells (PBMC) were measured to antigens of Chlamydia trachomatis in 29 subjects with severe conjunctival scarring and compared with responses from 29 age-, sex-, and location-matched controls from a trachoma-endemic area in The Gambia. Proliferative and interferon gamma responses were measured against chlamydial elementary bodies, recombinant major outer membrane protein, and affinity-purified recombinant chlamydial heat shock protein 60. Serum antibody levels in response to these antigens were also compared between the two groups, and the presence of ocular chlamydial antigen and DNA was assessed. PBMC from donors with conjunctival scarring had reduced proliferative responses to chlamydial antigens but not control antigens or mitogens compared with matched controls. By contrast, subjects with scarring had significantly higher levels of anti-chlamydial IgG antibody compared with matched controls and were more likely to be harboring ocular chlamydial antigen or DNA (or both). PMID- 8245541 TI - Invasive pneumococcal disease in central Oklahoma: emergence of high-level penicillin resistance and multiple antibiotic resistance. Pneumococcus Study Group. AB - Relatively penicillin-resistant pneumococci have caused 10% of invasive pneumococcal disease in central Oklahoma during the last decade, but almost no high-level penicillin or other antibiotic resistance has been described. This study evaluated antibiotic susceptibility and serotype distribution in invasive pneumococcal disease in the Oklahoma City metropolitan area (1990 population 848,000). A total of 144 cases of invasive infection was collected in 1 year (17 with meningitis, 120 with other bacteremic infections, and 7 with other invasive infections), for a rate of 16.9/100,000 (95% confidence interval [CI], 14.0 19.5). For the population aged > or = 60, invasive pneumococcal disease rates were higher among nursing home residents (352/100,000) than among nonresidents (25.6/100,000; relative risk, 13.7; 95% CI, 7.7-24.7). Antibiotic-resistant organisms caused 19.4% of the cases: relative penicillin resistance, 7.6%; high level penicillin resistance, 1.4% (2 cases), and 11% resistance to erythromycin, trimethoprim-sulfamethoxazole, or both, with 5% sharing both resistances plus a MIC of penicillin of 0.06 microgram/mL. PMID- 8245542 TI - Safety and immunogenicity of live oral cholera vaccine candidate CVD 110, a delta ctxA delta zot delta ace derivative of El Tor Ogawa Vibrio cholerae. AB - The current pandemic of cholera is caused primarily by Vibrio cholerae O1 of the El Tor biotype. Live attenuated classical biotype V. cholerae vaccine strains prevent severe and moderate cholera due to either biotype in challenged volunteers but may provide less protection against mild cholera due to El Tor organisms. CVD 110, a new ctxA-deleted vaccine strain derived from an El Tor Ogawa parent, lacks zona occludens toxin (Zot), accessory cholera enterotoxin (Ace), and hemolysin/enterotoxin. Ten healthy adult volunteers were given 10(8) cfu of CVD 110 with buffer; 7 developed diarrhea (mean stool volume, 861 mL). Vaccine organisms were shed in stool by all vaccines and were recovered from duodenal fluid in three-quarters of vaccinees. After vaccination, the geometric mean peak reciprocal vibriocidal titer among vaccinees was 17,829. CVD 110 is a powerful immunogen but insufficiently attenuated despite the absence of known potential enterotoxins of V. cholerae. Another unrecognized toxin or colonization alone may be responsible for diarrhea after ingestion of this strain. PMID- 8245543 TI - Direct DNA amplification and restriction pattern analysis of Helicobacter pylori in patients with duodenal ulcer and their families. AB - Sera from 14 patients with duodenal ulcer and their families were tested for IgG antibodies to Helicobacter pylori. Fourteen serologically negative patients and their families served as controls. Index patients and their family members who were serologically positive were advised to undergo endoscopic biopsy. Gastric biopsy tissues were subjected to HaeIII restriction analysis of nested polymerase chain reaction products of the urease gene. By serology, 28 (49.0%) of 57 in index families and 11 (27.5%) of 40 in control families tested positive. A higher prevalence rate was found in children of index patients (11/31, 35.5%) than in those of control patients (1/18, 5.6%; P < .05). On DNA analysis, 11 patterns were found in 13 patients, and 6 families underwent endoscopy. Children in 5 families exhibited identical patterns to those of their siblings and, in 3 of the 5 families, identical to the pattern of 1 of the parents. These results suggest that parent-to-child transmission and common infection source are probable causes of intrafamilial clustering of H. pylori. PMID- 8245544 TI - Diagnosis of Helicobacter pylori infection in a developing country: comparison of two ELISAs and a seroprevalence study. AB - Serology to detect antibodies to Helicobacter pylori is not frequently used as a diagnostic tool in developing countries. When compared to a commercial ELISA, an ELISA constructed and validated in Thailand had a higher sensitivity (98% vs. 85%), specificity (76% vs. 66%), and negative predictive value (97% vs. 76%) for the detection of H. pylori infection among 104 patients with dyspepsia evaluated by endoscopy. The positive predictive value was 88% for both tests. Serum antibody levels fell significantly 5-8 months after eradication of infection in 8 Thai patients (P = .009). By 8 years of age, > 50% of Thai persons living in urban and rural locations were seropositive. The low negative predictive value of the commercial ELISA limits the usefulness of this assay as a diagnostic tool in Thailand and suggests a need to reevaluate H. pylori serologic tests when used in populations living in developing countries. PMID- 8245545 TI - Evidence of previous infection with Mycobacterium avium-Mycobacterium intracellulare complex among healthy subjects: an international study of dominant mycobacterial skin test reactions. AB - Skin tests with 0.1 mL of intermediate-strength Mycobacterium tuberculosis purified protein derivative (PPD) and 0.1 mL of Mycobacterium avium sensitin were conducted on 484 healthy subjects from diverse geographic sites. Reactions of > or = 5 mm to one antigen that exceeded the reaction to the other by > or = 3 mm were considered M. avium- or PPD-dominant. PPD-dominant reactions were more frequent at sites where routine Bacille Calmette-Guerin immunization is done or where there are high rates of tuberculosis: New Hampshire, 2%; Boston, 7%; Finland, 14%; Trinidad, 26%; and Kenya, 28%. However, rates of M. avium-dominant reactions ranged from 7% to 12% at all sites. Analysis of dominant reactions based on a more stringent 10-mm minimum reaction size showed similar trends. These data suggest that exposure to MAC is similar in developed and developing countries but that broad mycobacterial immunity is greater in developing countries and may contribute to the lower rates of disseminated MAC infections in AIDS in these areas. PMID- 8245546 TI - Low serum levels of oral antimycobacterial agents in patients with disseminated Mycobacterium avium complex disease. AB - Twenty-seven human immunodeficiency virus-infected patients with disseminated Mycobacterium avium complex disease who were treated with oral antimycobacterial agents (clofazimine, ciprofloxacin, ethambutol, and rifampin) were studied to evaluate the usefulness of monitoring serum drug concentrations and testing in vitro susceptibility of M. avium complex (MAC) isolates. Twenty patients tolerated treatment with three or four antimycobacterial agents for at least 8 weeks; mycobacteremia was eradicated in 7 (35%). The in vitro susceptibilities of MAC isolates to antimycobacterial agents were similar for these 7 and for the 13 who did not respond to antimycobacterial treatment. Serum drug levels were below the expected range in 6 of the 7 whose mycobacteremia was cleared and in 9 of the 13 nonresponders (P = .41). These low serum concentrations of antimycobacterial drugs may be due to impaired drug absorption in patients with AIDS and disseminated MAC disease. PMID- 8245547 TI - Defective antifungal activity of monocyte-derived macrophages from human immunodeficiency virus-infected children against Aspergillus fumigatus. AB - Invasive aspergillosis recently has been encountered in adults and children with human immunodeficiency virus (HIV) infection even without known risk factors, such as neutropenia or corticosteroid therapy. Macrophages play a significant role in the host defenses against Aspergillus organisms by ingesting conidia and preventing their germination to hyphae. The antifungal activity of peripheral blood monocyte-derived macrophages (MDM) from 19 HIV-infected children was compared with that of 16 normal controls. The phagocytic activity of patients' MDM, measured as percentage of phagocytosis, was significantly decreased compared with normal donors (P = .014). In addition, the inhibitory activity of MDM on germination of intracellular A. fumigatus conidia was significantly impaired in patients compared with normal controls (P = .016). There was no significant difference in the defects between patients with lower or higher CD4 lymphocyte counts. Impairment of antifungal activity of macrophages may contribute to the susceptibility of HIV-infected patients to aspergillosis. PMID- 8245548 TI - Anticryptosporidial activity of lasalocid and other ionophorous antibiotics in immunosuppressed rats. AB - The anticryptosporidial activities of three ionophorous antibiotics were assessed in dexamethasone-immunosuppressed rats infected with Cryptosporidium parvum. When administered prophylactically, lasalocid prevented infection in a dose-dependent manner, whereas monensin and salinomycin were ineffective. Therapeutically, lasalocid eliminated established overt infections of the intestine, although infection persisted in the common bile duct and intestinal infection recurred after lasalocid treatment was stopped. These findings suggest that lasalocid is a potentially useful anticryptosporidial agent but that long-term continuous administration may be necessary in the immunocompromised host. PMID- 8245549 TI - Small subunit rRNA sequence of Enterocytozoon bieneusi and its potential diagnostic role with use of the polymerase chain reaction. AB - In the past several years, microsporidia have become recognized as another important group of opportunistic infections of immunocompromised patients, especially those with AIDS. Enteric infections with the noncultivatable microsporidian parasite Enterocytozoon bieneusi have been diagnosed from AIDS patients with chronic diarrhea, malabsorption, and wasting. The incidence of infection and mechanism of transmission of these organisms in humans is unknown. Several recent tests for human pathogens have been developed using rRNA genes as diagnostic probes. Using the polymerase chain reaction and conserved regions of the small subunit rRNA (SSU-rRNA) gene, the SSU-rRNA gene of E. bieneusi was successfully cloned and subsequently sequenced. Amplification of E. bieneusi rRNA could be demonstrated from intestinal biopsies from HIV-1-infected patients infected with E. bieneusi but not from intestinal biopsies from noninfected patients. This cloned SSU-rRNA gene was used to develop improved probes for detection of E. bieneusi in tissue of infected patients. PMID- 8245550 TI - Schistosoma mansoni egg attachment to cultured human umbilical vein endothelial cells: an in vitro model of an early step of parasite egg excretion. AB - To model the attachment of Schistosoma mansoni eggs to the endothelium of the mesenteric vasculature, the interaction between S. mansoni eggs and cultured human umbilical vein endothelial cells (HUVEC) in vitro was investigated. S. mansoni eggs rapidly attached to monolayers of both HUVEC and bovine aortic endothelial cells but more slowly to monolayers of cultured fibroblasts and smooth muscle cells. While both native and glutaraldehyde-fixed eggs attached equally well to HUVEC, eggs attached only to live, metabolically active HUVEC. Attachment was enhanced by both serum and plasma factors. In addition, platelet release products increased egg attachment by 75%. Preincubation of S. mansoni eggs with soluble egg antigens promoted attachment; in contrast, preincubation of HUVEC with the antigens inhibited attachment. These results suggest that interaction of S. mansoni eggs with HUVEC is an active process that can be modulated by molecules secreted by the egg and by platelets during egg extravasation. PMID- 8245551 TI - Comparability and reliability of ELISA, immunofluorescence, and indirect hemagglutination assays for Trypanosoma cruzi and Trypanosoma rangeli. AB - Serum samples and filter paper spot samples from 48 adults were assayed in three laboratories for Trypanosoma cruzi and Trypanosoma rangeli by ELISA, immunofluorescence (IF), and indirect hemagglutination (HA) in a controlled, blind experiment. IF (serum samples) attained higher levels of comparability necessary for validity than did ELISA (serum samples) in these laboratories. ELISA was inferior to IF and HA (spot samples) in testing for T. cruzi. ELISA was not reliable (intrasubject r = .51) and was not in agreement between two laboratories (kappa = .09). The experiment indicates that ELISA is not the best choice of assay for T. cruzi in epidemiologic surveys. Assays for T. rangeli by ELISA and IF lacked comparability to the extent that those two techniques could not be considered useful for T. rangeli. PMID- 8245552 TI - Detection of Toxoplasma gondii by competitive DNA amplification of bronchoalveolar lavage samples. AB - The prevalence of pulmonary toxoplasmosis was assessed by a prospective analysis of 144 bronchoalveolar lavage (BAL) samples using competitive polymerase chain reaction (PCR) to avoid false-negative results due to PCR techniques. Six samples were excluded because they contained amplification reaction inhibitors. None of the samples from the 37 immunocompetent patients and only 1 sample (1.7%) from the 59 immunocompromised patients without human immunodeficiency virus infection were PCR-positive. In contrast, Toxoplasma gondii DNA was found in 6 (14%) of 42 samples from patients with AIDS. All 6 patients had < or = 40 CD4 cells/microL and anti-Toxoplasma antibodies, and 5 had other sites of Toxoplasma infection. Six other AIDS patients who had received treatment for cerebral toxoplasmosis were PCR-negative. Thus, pulmonary toxoplasmosis is frequent in AIDS patients who have other sites of Toxoplasma infection and low CD4 lymphocyte counts and who are not receiving prophylaxis. PMID- 8245553 TI - Efficacy of gloves in reducing blood volumes transferred during simulated needlestick injury. AB - This study was designed to evaluate factors that affect blood volumes transferred to skin during simulated needlestick injuries in an in vitro paper prefilter model and an ex vivo porcine tissue model. The effect of needle type and size, penetration depth, and glove use on the volume of radiolabeled blood transferred was determined in each model. Blood volumes ranged from 0.47 +/- 0.26 microL (30 gauge needle, 0.5-cm depth, in vitro model) to 5.88 +/- 1.45 microL (18-gauge needle, 2.0-cm depth, in vitro model). Needle size and penetration depth were significantly associated with transfer volume. Glove material reduced the transferred blood volume by 46%-86% in both models. Transfer volumes were within the same order of magnitude for all conditions. Hence, virus titer in the source blood may be a better predictor of needlestick infectivity than is exposure volume. Regardless, gloves may exert some protective effect and should be worn whenever needles are handled. PMID- 8245554 TI - Protection against lethal endotoxemia by monoclonal antibodies. PMID- 8245555 TI - Aminoglycoside toxicity. PMID- 8245556 TI - Maternal measles and rubella antibody levels and serologic response in infants immunized with MMR II vaccine at 12 months of age. PMID- 8245557 TI - Evaluation of a bovine-human rotavirus reassortant vaccine in infants. PMID- 8245558 TI - Spontaneous lymphocyte proliferation in human T lymphotropic virus type II infection is associated with increased provirus load. PMID- 8245559 TI - Treatment trials for primary human immunodeficiency virus type 1 infection. PMID- 8245560 TI - Infections caused by imipenem-resistant Acinetobacter calcoaceticus biotype anitratus. PMID- 8245561 TI - Activity of doxycycline against preerythrocytic malaria. PMID- 8245562 TI - United we stand, divided we fall. PMID- 8245563 TI - Augmentation genioplasty with hydroxyapatite blocks. A case report. PMID- 8245564 TI - One-stage closure of isolated cleft palate with the Veau-Wardill-Kilner V-Y push back method or the Cronin modification. Cephalometric comparison of nasopharynx. AB - The craniofacial structure of 116 consecutive patients with isolated cleft palate was studied by lateral cephalograms at 17-20 years of age. One-stage soft- and hard-palate closure had been done with the Veau-Wardill-Kilner or the Cronin mucoperiosteal palatal V-Y push-back method at the mean age of 1.8 years. The patients treated with the Veau-Wardill-Kilner method showed larger sagittal depths of the bony nasopharynx and nasopharyngeal airway but smaller thickness of the soft tissue on the posterior nasopharyngeal wall. The sagittal depth of the nasopharyngeal airway was larger in patients who had had secondary palatal operations. The effect of sex was significant, males having larger dimensions. PMID- 8245565 TI - Surgical considerations for endosseous implants in the craniofacial region: a 3 year report. AB - From 1987 to 1990, 28 patients were treated with titanium implants (Branemark flange fixtures) in the craniofacial area for the support of craniofacial prosthesis or anchorage of bone-conduction hearing aids in the Orofacial Implant Center at the University of California, Los Angeles (UCLA). A total of 88 implants were placed, of which 22 were placed in previously irradiated areas. Twenty-three of the implants were placed in the orbital area, 37 in the mastoid area, 20 in the nasal area, and eight in various other craniofacial regions. Seven implants were lost in four different patients. The proper location and positioning of implants in specific areas of the craniofacial regions, as well as soft-tissue complications and management, are discussed. PMID- 8245566 TI - Cervical and mediastinal emphysema secondary to mandible fracture: case report and review of the literature. AB - Cervical and mediastinal emphysema is a rare but serious, life-threatening complication associated with mandibular fractures secondary to high-impact trauma. A case is reported in which a 24-year-old white man involved in a motor vehicle accident presented with an isolated mandibular fracture, cervical emphysema, and pneumomediastinum. A review of the literature is presented, and relevant anatomy and management are discussed. PMID- 8245567 TI - Clinical impact of synchronous and metachronous malignancies in patients with oral cancer. AB - In a retrospective study, 20 of 379 patients (5.2%) with cancer of the oral cavity developed second malignant tumors (SMT) at various sites. The diagnosis of SMT was made with an average delay of 49.2 months. In view of the increasing incidence of SMT after curative treatment of oral cancer, the use of chemopreventive drugs should be considered. PMID- 8245568 TI - p53 and Ki 67 expression in preneoplastic and neoplastic lesions of the oral mucosa. AB - Some lesions of the oral mucosa such as leukoplakia and erythroplakia may develop into squamous cell carcinoma (SCC). At present, however, there is no method available to predict malignant transformation. It is known that the grade of dysplasia is related to the potential malignant development, but this is unreliable as the only indicator. In 64 hyperplastic lesions and 85 SCC of the oral mucosa, a correlation between the expression of the mutated tumor-suppressor gene p53 and the dysplasia of the lesions was found. Ki 67 was used as a proliferation marker. The results imply that expression of mutated p53 is an indicator for potential malignant development in benign lesions of the oral mucosa. PMID- 8245569 TI - Acinic cell carcinoma arising in a parotid lymph node. AB - The case of a 37-year-old woman with primary acinic cell carcinoma arising in an intraparotid lymph node is presented. The patient is free of disease 20 months after superficial parotidectomy. This is probably the first histologically documented case of acinic cell carcinoma arising from intranodal salivary gland tissue. Awareness of possible malignant alteration of ectopic salivary gland tissue in lymph nodes is essential. PMID- 8245570 TI - Septic arthritis of the temporomandibular joint: review of the literature and report of two cases in children. AB - Septic arthritis of the temporomandibular joint (TMJ) has a high morbidity, is infrequently reported, and has been described almost exclusively in adults. We present two cases of septic arthritis of the TMJ that occurred in children after minor blunt trauma. Literature related to septic arthritis of the TMJ was reviewed, and a composite list of cases was constructed. The most common causes were various infections of the head and neck, rheumatic joint disease, and iatrogenesis. Pathogens may gain access to the TMJ by several routes. Patients typically present with an acute, tender, monarticular arthritis with associated swelling and erythema. Malaise, nausea, and vomiting may also be present. Traumatic effusions, fractures, and neoplasms may present in a similar fashion, and mimic TMJ septic arthritis. Staphylococcus aureus is the most commonly reported pathogen and often causes permanent joint damage. Aspiration and analysis of joint fluid, as well as blood chemistry, imaging studies, and clinical impression, may assist in the diagnosis. Timely diagnosis and treatment are essential for a successful outcome; therapy should include antimicrobial agents, adequate drainage, and resting of the joint. Complications include spread of infection, postinfectious bony changes, and fibrous (or bony) ankylosis of the temporomandibular joint. PMID- 8245571 TI - Epidermal nevus syndrome with maxillary involvement. AB - A female patient with epidermal nevus syndrome is reported. There were linear epidermal nevi, hemihyperplasia of the limbs and tongue, macrocephaly, several ophthalmic malformations, and multiple radiolucent lesions in the limbs and sacroiliac region. At age 14 years, she developed a giant cell granuloma of the maxilla. PMID- 8245572 TI - Binding pattern of eight different lectins in healthy subjects and patients with dysplastic and malignant lesions of the oral cavity. AB - The present investigation was undertaken to assess the binding characteristics of eight different lectins to normal oral mucosa (11 cases), leukoplakia with varying degrees of dysplasia (five cases), and oral mucosal squamous cell carcinoma (12 cases) by the use of biotinylated lectins and avidin biotin peroxidase complexes. The lectins employed were soybean (SBA), peanut (PNA), Dolichos biflorus (DBA), concanavalin A (Con A), wheat germ (WGA), Ulex europaeus (UEA), Ricinus communis (RCA), and Lotus tetragonolobus (LTA). It was observed that SBA, DBA, WGA, UEA, RCA, and LTA showed very strong to strong binding in healthy oral mucosa but no or very weak binding in squamous cell carcinoma. On the contrary, PNA showed weak binding to normal mucosal epithelial cells but showed strong binding to malignant cells. The dysplastic mucosa had an intermediate binding pattern. The lectin Con A was not bound at all or seen in very low concentration in the malignant cells and dysplastic epithelium, but it showed weak binding in the normal mucosa. Hence, we conclude that lectins may be utilized as probes to determine the dysplastic and malignant status of the oral mucosal epithelium. PMID- 8245573 TI - The role of the periosteum in osteointegration of hydroxyapatite granules. AB - The role of the periosteum in the osteointegration of hydroxyapatite (HA) granules was studied in the skull and tibial bone of eight adult Gottingen minipigs. Calcein and tetracycline were used as labeling stains to determine new bone formation. Depending on the condition of the periosteum, three different patterns of osteointegration of HA were found. PMID- 8245574 TI - Service-mix of oral and maxillofacial surgeons in Australia and New Zealand. AB - This study aimed to investigate the service-mix of oral and maxillofacial surgeons and its association with patient age, sex, and referral sources. Oral and maxillofacial surgeons in Australia and New Zealand were surveyed (n = 97; 71.9% response rate), and they supplied service-mix data from a week-long log of services provided to 5770 patients. Dentoalveolar services comprised 67.0% of services. Subcategories of service-mix showed high levels of removal of unerupted teeth (44.8%) and multiple extractions (10.0%), with soft-tissue lesions (5.0%), TMJ surgery (4.7%), mandible trauma (4.6%), and simple extraction (4.4%) the next categories in order of percentage. Variations in service-mix were observed for patient age and sex, among referral sources, and between private and public practice. PMID- 8245575 TI - A consensus approach to projections of the supply of oral and maxillofacial surgeons in Australia. AB - The analysis presented involves projections of the future requirements for the oral and maxillofacial surgeon labour force in Australia under a range of different recruitment scenarios, followed by a consensus-seeking exercise with the Australian and New Zealand Association of Oral and Maxillofacial Surgeons (ANZAOMS) Council in an attempt to produce estimates of the most probable future requirement for oral and maxillofacial surgeons. The resulting consensus estimates would result in the maintenance of the present population-to-surgeon ratio. Different premises or beliefs underlying the projections were viewed as counterbalancing each other. PMID- 8245576 TI - Stanford 3D hyperthermia treatment planning system. Technical review and clinical summary. AB - In the field of deep regional hyperthermia cancer therapy the Sigma 60 applicator of the BSD-2000 Hyperthermia System is one of the most widely used devices. This device employs four independent sources of radiofrequency electromagnetic energy to heat tumour sites deep within the body. The difficulty in determining the input parameters for the four sources has motivated the development of a computer based three-dimensional (3D) treatment planning system. The Stanford 3D Hyperthermia Treatment Planning System has been in clinical use at Stanford Medical Center for the past 2 years. It utilizes a patient-specific, three dimensional computer simulation to determine safe and effective power deposition plans. An optimization programme for the selection of the amplitudes, phases and frequency for the sources has been developed and used in the clinic. Examples of the application of the treatment planning for hyperthermia treatment of pulmonary, pelvic, and mediastinal tumours are presented. Methods for quantifying the relative effectiveness of various treatment plans are reviewed. PMID- 8245577 TI - Thermoradiotherapy with combined interstitial and external hyperthermia in advanced tumours in the head and neck with depth > or = 3 cm. AB - Advanced tumours in the head and neck 3-6 cm depth are too deep to be completely heated by external 915 MHz microwaves. A preliminary study was performed using interstitial plus external hyperthermia combined with external beam radiation therapy to heat tumours to depths > or = 3 cm. Nine advanced metastatic lesions of squamous cell carcinoma located in the head and neck were treated between 1987 and 1990 with the combined hyperthermia technique and radiation doses of 38-60 Gy (mean of 49 +/- 3 Gy). The mean tumour volume was 58 +/- 9 (SE) cm3 (range 24-94 cm3) with a mean tumour depth of 3.9 +/- 0.3 cm (range 3-5.5 cm). The deeper aspects of the tumour were heated by interstitial 915 MHz microwave antennas and the superficial aspects heated by external 915 MHz applicators. A single plane of polyurethane closed-end catheters, 16 Ga, were inserted under local anaesthesia approximately 1.5-2 cm apart in parallel arrays at the base of a lesion behind the sternomastoid muscle, or an equivalent site in a dissected neck, extending forward and angled deeply no more than 15 degrees. Hyperthermia was administered twice weekly immediately after radiation therapy in a mean of 5.3 +/- 0.7 external heat sessions (range 3-7) and a mean of 3.5 +/- 0.6 interstitial heat sessions (range of 1-6). Interstitial hyperthermia was usually administered in alternating sessions with external hyperthermia, but in some patients all of the sessions of one modality were administered followed by all of the sessions of the other modality. In no case were both interstitial and external heatings performed on the same day. Surface thermometers were used to monitor skin temperature during external hyperthermia sessions. Results showed that by 8 weeks after completion of treatment, six lesions exhibited a complete response (67%) and three a partial response (33%). One of the partial responses continued to regress and became a complete response (78% complete response). The recurrence rate in complete responders was 14% (1/7) with time to recurrence of 7.7 months. Six lesions were recurrence-free at last follow-up of 21.3 +/- 8.8 months. Skin reactions were absent in four fields (44%), erythema was noted in five (56%) and thermal blistering in one (11%). Ulceration occurred only in association with tumour breakdown when the skin was infiltrated by tumour (three patients, 33%).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8245578 TI - 27 MHz hybrid evanescent-mode applicators (HEMA) with flexible heating field for deep and safe subcutaneous hyperthermia. AB - A new class of low-frequency electromagnetic applicators for hyperthermic treatment of superficial and subcutaneous tissues is described. These applicators employ an air-filled waveguide segment which is operating below the cut-off frequency, the evanescent modes of which are energized by suitable exciters to produce model field components. Direct radiators are integrated into the waveguide to generate additional direct field components. All field components may be combined in different power level ratio, phase, and orientation, to provide a composite heating field exhibiting a large variety of field sizes, shapes, and penetration features. The composite field emerging from the waveguide aperture propagates within the tissue to be heated through a non-critical air gap. These versatile heating devices appear of potential interest to heat a variety of deep and localized subcutaneous tissues to therapeutic temperatures without injury to access fat layers of substantial thickness. PMID- 8245579 TI - Heating patterns of the Helios ultrasound hyperthermia system. AB - Thermal dosimetry studies of the Helios ultrasound system were performed in a tissue-equivalent phantom and in porcine tissues in vivo. SAR distributions in the phantom demonstrate the adjustability of the power deposition pattern from the 50 cm diameter annular array of 30 transducers. Examples are given of well localized SAR patterns (< 3 cm diameter) in the phantom, and of peripherally enhanced (doughnut-shaped) SAR patterns up to 10 cm in diameter at the focal plane. In vivo trials in porcine thigh muscle demonstrate the ability of the Helios system to produce regions of therapeutic heating (42-45 degrees C) as small as 20 cm3 and as large as 200 cm3 at depths of greater than 10 cm at a frequency of 0.5 MHz. PMID- 8245580 TI - Temperature and SAR measurements in deep-body hyperthermia with thermocouple thermometry. AB - Multisensor (7-14) thermocouple thermometry is used at our department for temperature measurement with our 'Coaxial TEM' regional hyperthermia system. A special design of the thermometry system with high resolution (0.005 degrees C) and fast data-acquisition (all channels within 320 ms) together with a pulsed power technique allows assessment of specific absorption rate (SAR) information in patients along catheter tracks. A disadvantage of thermocouple thermometry, EM interference, is almost entirely eliminated by application of absorbing ferrite beads around the probe leads. We investigated the effect of remaining disturbance on the temperature decay after power-off, both experimentally in phantoms and in the clinic, and with numerical simulations. Probe and tissue characteristics influence the response time tau dist of the decay of the disturbance. In our clinical practice a normal pulse sequence is 50 s power-on, 10 s power-off: a response time longer than the power-off time results in a deflection of the temperature course at the start. Based on analysis of temperature decays correction of temperature is possible. A double-pulse technique is introduced to provide an initial correction of temperature, and fast information about accuracy. Sometimes disturbance with a relatively long response time occurs, probably due to a bad contact between probe, catheter and/or tissue. Thermocouple thermometry proved to be suitable to measure the SAR along a catheter track. This is used to optimize the SAR distribution by patient positioning before treatment. A clinical example illustrates this. PMID- 8245581 TI - Effects of pre-heating on cis-diamminedichloroplatinum(II)-hyperthermia-induced tumour growth depression of transplantable human oesophageal cancer to nude mice. AB - The effects of the combination of cis-diamminedichloroplatinum(II) (CCDP) and hyperthermia on tumour growth were examined using transplantable human oesophageal cancer (ESO-2), having a histological type of moderately differentiated squamous cell carcinoma, in nude mice. Eighteen days after the inoculation of the tumour fragment into the subcutaneous tissue of the right hind foot, the treatment of CDDP and/or hyperthermia was performed and the antitumour effect was evaluated 21 days after the treatment. The combination of 4 mg/kg of CDDP and 43 degrees C heating for 30 min effectively depressed tumour growth in comparison with the individual treatment. The mean relative tumour weight of the combination group at 3 weeks after the treatment was 15% of that of the control group without treatment. On the other hand, pre-heating at 42 degrees C for 30 min did not influence the inhibition of tumour growth by CDDP alone or the concentration of CDDP in tumour. When pre-heating at 42 degrees C for 30 min was performed at 6 or 12 h prior to the combined treatments of 2 mg/kg of CDDP and 43 degrees C hyperthermia for 30 min, however, tumour growth depression by CDDP hyperthermia was diminished. When pre-heating was performed 4 days prior to CDDP hyperthermia, however, tumour growth depression occurred. These results showed that thermotolerance of tumour cells induced by pre-heating diminished hyperthermic potentiation for cytotoxicity of CDDP, though the thermotolerance did not affect the cytotoxicity of CDDP. PMID- 8245582 TI - Effects of hyperthermia on avian red blood cells. AB - Our primary goal was to develop a model for studying the effects of heat on terminally differentiated cells. Using nucleated chicken red blood cells (RBC) as the model, heat dose-response and thermotolerance were investigated. A new haemolytic assay was developed to score them. Heating chicken RBC for as long as 3 h at 51.5 degrees C resulted in only small amounts of haemolysis immediately after heating. When haemolysis was scored 1-2 days after heating, heat-induced haemolysis increased with heating temperature and duration. Heat shock of 30-90 min at 42.6 degrees C just before heat challenge of 40 min at 51.5 degrees C, or heat shock of 15 min at 43.1 degrees C with incubation of 0-2 h at 35-37 degrees C prior to the same heat challenge induced thermotolerance, but the levels of heat resistance achieved were different. Similar experiments were performed using potassium leakage as the endpoint. Potassium leakage was measurable immediately after heating, and it increased with heating duration, but there was no correlation between potassium leakage and haemolysis. Potassium leakage was not a suitable criterion for measuring thermotolerance. PMID- 8245583 TI - Ultrastructural changes in the rat sciatic nerve after local hyperthermia. AB - The rat sciatic nerve was heated over a length of 5 mm for 30 min at 43, 44 or 45 degrees C. Morphological changes were not observed after heating at 43 degrees C. Treatment at 44 degrees C resulted in endoneurial oedema and mild vascular changes, such as contraction and vacuolization of endothelial cells and thickening of the media of the larger vessels. Within 1 week several demyelinated axons were observed. The first changes after heating at 45 degrees C included oedema, blood vessel occlusion and severe endothelial cell damage. Axonal changes, e.g. the accumulation of cell organelles, appeared 8 h after treatment; 24 h after treatment most axons and myelin sheaths showed degenerative changes. Absence of blood flow in the heated area of the nerve was shown 2 h after heating at 45 degrees C. We conclude that hyperthermic treatment directly affects endothelial cells and myelin sheaths in the rat sciatic nerve. Axons degenerate most probably as a consequence of ischaemia. PMID- 8245584 TI - Manipulation of intra- and extracellular pH in spontaneous canine tumours by use of hyperglycaemia. AB - We evaluated the use of hyperglycaemia to reduce tumour pH in dog with spontaneous tumours. Dogs were randomized to two groups: control and glucose. Intravenous administration of 20% glucose was used to induce and maintain hyperglycaemia. Extra- and intracellular tumour pH were measured using interstitial pH microelectrodes and 31P-MRS, respectively. During the administration of glucose, the mean (+/- SEM) blood glucose concentration was 419.8 (+/- 32.8) and 121.1 (+/- 8.0) mg/dl for the glucose and control groups, respectively. The mean extracellular tumour pH before and following 90 min of hyperglycaemia was 7.15 (+/- 0.08) and 7.15 (+/- 0.09). During consecutive measurements, intracellular tumour pH did not change significantly for the control group or the group subjected to hyperglycaemic manipulation. In contradistinction to previous rodent studies, our results demonstrate that hyperglycaemia alone is not sufficient to manipulate either intra- (pHi) or extracellular (pHe) hydrogen ion concentration in spontaneous canine soft tissue tumours. PMID- 8245585 TI - Thermal dose and secondary tumour cell death. AB - Both primary and secondary tumour cell death may occur in clinical hyperthermia. The equation usually used for calculation of thermal dose takes only the primary cell death into consideration. We propose that the thermal dose equation should be reassessed; contributions from secondary cell death should also be included. The secondary cell death is governed mainly by the temperature distribution during treatment, the arteriolar density distribution in the tumour and the heat sensitivity of the arterioles. Increased thermal dose and hence increased tumour treatment response may result if hyperthermic treatments are designed to maximize the secondary cell death. Massive secondary cell death may be achieved by inducing hot spots in tumour areas with high arteriolar density, identified in pretreatment tumour angiograms. PMID- 8245586 TI - Temperature errors using multi-sensor thermocouple probes with a common constantan wire. PMID- 8245587 TI - [The clinical significance of retroperitoneal lymph node metastasis and lymphadenectomy on 3-year survival in ovarian carcinoma]. AB - Fifty-two patients with primary epithelial ovarian cancer received surgical treatment and chemotherapy at the Hospital of Shiga University of Medical Science and retroperitoneal lymph node metastases were evaluated clinico-pathologically. Forty patients underwent paraaortic and pelvic lymphadenectomy, although 12 patients with stage IIIc underwent no lymphadenectomy. In the patients with lymphadenectomy, stage I and II groups had 100% of 3-year survival, stage III group, 40.0% and stage IV group, 0.0%. A series of 27 patients with stage IIIc diagnosed from findings in the abdominal cavity had cytoreductive surgery and chemotherapy including cisplatinum, and were evaluated for 3-year survival. No significant difference in the survival rate was found between the stage IIIc groups with and without para-aortic and pelvic lymphadenectomy. The most important prognostic factors for 3-year survival were retroperitoneal lymph node metastases, complete remission, and residual tumor at second surgery. A three year survival analysis confirmed that para-aortic and pelvic lymphadenectomy is not an important procedure for the treatment for stage III ovarian cancer. However, retroperitoneal lymph node metastases was one of the most important prognostic factors. PMID- 8245588 TI - [Ultrasonographic study of low-lying placenta and its clinical significance]. AB - Low-lying placenta is reported to cause PROM, preterm labor, and abnormal bleeding during pregnancy and labor, but it has not been clearly defined how low the placenta has to be implanted to categorize it as "low-lying". In the present investigation, using ultrasonography, the distance between the placental implantation site and the internal cervical os in relation to gestational outcome was investigated to clarify the diagnosis and clinical significance of low-lying placenta. 1. Placental implantation site in normal gestation 1) In none of the cases at week 28 of gestation or later, the distance was 2cm or less between the lower edge of the placenta and the internal cervical os. At week 32 or later, the distance was 3cm or less in none. 2) At week 36 or later, it was 5 cm or more in all cases. 2. Low-lying placenta in association with gestational or labor outcome When "low-lying placenta" was tentatively defined as the placenta implanted within 2cm from the internal os and "normal placenta" as one implanted 2.1 cm or more from the internal os, the following findings were obtained. 1) The incidence of low-lying placenta was 2.8% (49/1,771) at gestational weeks 20 < or = - < 24, 0.96% (17/1,771) up to gestational week 29, and 0.73% (13/1,771) up to 1 week prior to labor. 2) The incidence of bleeding during gestation, preterm labor, PROM and emergency C/S was higher in the 13 cases with low-lying placenta remaining until labor than in 1,749 cases (p < 0.05) normally sited.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245589 TI - [The efficacy of prophylactic antibiotic and tocolytic therapy for premature rupture of the membranes--a prospective randomized study]. AB - A comparative study in patients with premature rupture of the membranes (PROM) from 25 to 34 weeks of gestation was carried out, prospectively. Group 1 (34 patients) was given aggressively intrauterine therapy including the administration of tocolytic agents (ritodrine and/or magnesium sulfate) and prophylactic antibiotics (AB-PC 2g/day). Group 2 (41 patients) was managed conservatively with bed rest only. At the time of admission to the study, there were no clinical signs of infection, fetal distress, or active labor in either group. All patients were delivered if the pregnancy had reached 35 weeks of gestation or later, had established labor, or developed evidence of chorioamnionitis or fetal distress. Prolongation for more than 72 hours was greater in group 1 than in group 2. There was no difference in the incidence of chorioamnionitis, postpartum endometritis, or placental infection in the groups. However, the incidence of a low Apgar score (7 < at 5 min), requiring artificial ventilation, and infection was more common in group 1. It is concluded that the use of antibiotics and tocolytics might make the management of PROM more complicated. PMID- 8245590 TI - [Clinical significance of magnetic resonance imaging (MRI) in evaluation of the extension of uterine cervical cancer]. AB - Magnetic resonance imaging (MRI) was performed in 62 patients with uterine cervical cancer and the preoperative MRI findings were compared with the pathological findings following surgery. The surgical stages of 62 patients were 2 at stage 0, 18 at stage Ia, 19 at stage Ib, 9 at stage IIa, 11 at stage IIb, 2 at stage IIIa, and 1 at stage IIIb. The MRI findings in the present study included: (1) the existence of a high intensity area (HIA) in the uterine cervix or minimum thickness of residual normal cervical tissue, (2) necrotic cavity, (3) pyometra, (4) irregular margin of the cervix, (5) parametrial invasion, (6) vaginal invasion, (7) bladder invasion, and (8) lymphnode enlargement. No HIA was observed in any patient with stage 0 or stage Ia, in 8 patients with stage Ib, and in 2 patients with stage IIa, while all patients with cancer tissues greater than 10mm in diameter had a HIA. In 39 patients in whom hysterectomy was undergone without conization, the findings of MRI correlated significantly (r = 0.929, p < 0.001) with the minimum thickness of residual normal cervical tissue by the pathological measurement. When the irregular margin of the cervix was regarded as a disruption of the cervical myometrium by cancer tissue, accuracy was 87%. Furthermore, the degrees of accuracy for parametrial invasion, vaginal invasion and bladder invasion were 92%, 90% and 94%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245591 TI - [Relationship among aromatase activity, estrogen receptor and progesterone receptor in ovarian tumors from postmenopausal women]. AB - Aromatase activity, as well as steroid receptors, has been demonstrated in ovarian tumors. Aromatase activity was detected in 35 tumors (81%), PR in 21 tumors (49%) and ER in 13 tumors (30%) out of 43 ovarian tumors (27 benign, 14 malignant and 2 granulosa cell tumors). Moreover, immunohistochemical study demonstrated aromatase cytochrome P-450 (P-450arom) in the tumor tissues. Aromatase activity was significantly greater in PR-positive tumors than in PR negative tumors (p < 0.002). There was a positive correlation between aromatase activity and the PR level (rs = 0.490, p < 0.001). Thirteen of 14 (93%) malignant tumors showed negative PR with low aromatase activity. However, the presence of ER was not correlated with aromatase activity or the presence of PR. There was a positive correlation among serum levels of estradiol, estrone, androstenedione and testosterone, whereas the serum steroid levels were not correlated with aromatase activity, the PR or ER of ovarian tumors. These results suggest that aromatase activity is correlated with PR in ovarian tumors of postmenopausal women. PMID- 8245592 TI - [Treatment of advanced ovarian cancer--is residual tumor diameter less than 2cm optimal?]. AB - The present study investigates whether residuum < 2cm is suitable for "optimal" by analyzing the impact of the size of the residuum on survival. Among 151 patients with stage III or IV ovarian cancer, 77 deemed "resectable" received debulking at initial laparotomy and received postoperative chemotherapy (PO group). The remaining 74 deemed "unresectable" had an exploratory laparotomy followed by neoadjuvant (NA) chemotherapy and subsequently received debulking (NA group). In the PO group, there was a significant difference in survival between patients with primary residuum < 2cm and > or = 2cm, and between < 0.5cm and > or = 0.5cm, respectively. Moreover, significant difference in survival was observed among each group with no residuum, macroscopic residuum but < 0.5cm, and 0.5cm < or = residuum < 2cm. The same trend was observed in the NA group. Altogether, a significant difference in survival was observed between patients with residuum < 0.5cm and those with 0.5cm < or = residuum < 2.0cm. In the PO interestingly, debulking the tumor to residuum < 2 cm did not influence the survival if postoperative chemotherapy was not effective. Thus, primary debulking of tumor to residuum < 2cm was significant when postoperative chemotherapy was effective. However, when we evaluated the residuum as an independent prognostic factor, residuum < 2 cm was insufficient but in view of the results of this study, < 0.5cm was judged to be suitable. PMID- 8245593 TI - [Glucose transport in human placental chorioepithelium]. PMID- 8245594 TI - [A case of ovarian serous papillary adenocarcinoma combined with malignant endosalpingiosis]. PMID- 8245595 TI - [The pregnancy and delivery of a women with bone marrow transplantation]. PMID- 8245596 TI - [Low-dose dopamine treatment for severe ovarian hyperstimulation syndrome]. PMID- 8245597 TI - [A case of pregnancy with antiphospholipid antibodies: serial measurement of flow velocity waveforms of uterine arteries]. PMID- 8245598 TI - [A case of bone marrow carcinomatosis complicated with DIC in a patient with uterine cervical carcinoma]. PMID- 8245599 TI - [Kidney failure. 1. Kidney failure--approach to background and diagnosis]. PMID- 8245600 TI - [Kidney failure. 2. Etiological mechanism and factors responsible for progression of the clinical course of acute renal failure]. PMID- 8245601 TI - [Kidney failure. 3. Mechanism and factors responsible for progression of the clinical course in chronic renal failure]. PMID- 8245602 TI - [Kidney failure. 4. Renal function of the aged and its evaluation (including obstructive nephropathy)]. PMID- 8245603 TI - [Acute renal failure. 1. Acute progressive glomerulonephritis syndrome]. PMID- 8245604 TI - [Acute renal failure. 2. Nephrosis with symptoms of renal failure- physiopathology, treatment, and prognosis]. PMID- 8245605 TI - [Acute renal failure. 3. Thrombotic microangiopathy]. PMID- 8245606 TI - [Acute renal failure. 4. acute renal failure caused by hypertension- physiopathology and management]. PMID- 8245607 TI - [Acute renal failure. 5. Renal infarction]. PMID- 8245608 TI - [Acute renal failure. 6. Renal failure caused by drugs or laboratory test procedures]. PMID- 8245609 TI - [Acute renal failure. 7. Postoperative acute renal failure--preventive management]. PMID- 8245610 TI - [Acute renal failure. 8. Hepatorenal syndrome--pathogenetic mechanism and treatment]. PMID- 8245611 TI - [Chronic progressive renal failure. 1. Diet therapy of chronic renal failure]. PMID- 8245612 TI - [Chronic progressive renal failure. 2. Chemotherapy of chronic active (progressive) glomerulonephritis]. PMID- 8245613 TI - [Chronic progressive renal failure. 3. The importance of antihypertensive agents and the treatment of chronic progressive renal failure]. PMID- 8245615 TI - [Chronic progressive renal failure. 5. Use of general medication in patients with reduced renal function]. PMID- 8245614 TI - [Chronic progressive renal failure. 4. Dietary instructions given to patients at various stages of diabetic nephropathy]. PMID- 8245616 TI - [Diagnosis and treatment of progressive renal failure: discussion]. PMID- 8245617 TI - [Case of pure red cell aplasia associated with granulo-lymphocytic leukemia successfully treated with cyclosporin A]. PMID- 8245618 TI - [Surgically treated case of abdominal aortic aneurysm associated with Weber Christian disease]. PMID- 8245619 TI - [Case of von Meyenburg complex with an echographic presentation suggestive of liver cirrhosis]. PMID- 8245620 TI - [Case of localized tuberculous hepatic abscess and tuberculous stomach ulcer discovered during treatment of tuberculous pleurisy]. PMID- 8245621 TI - [Case of anti-neutrophilic cytoplasm antibody-positive SLE succumbing to massive pulmonary hemorrhage]. PMID- 8245622 TI - [Onset and development glomerulonephritis and molecular biology]. PMID- 8245623 TI - [Recent progress in the study of the onset and recovery process of acute renal failure]. PMID- 8245624 TI - [Progress on the study of allergy and immunology in the field of internal medicine]. PMID- 8245625 TI - [Structure and function of the kidney]. PMID- 8245626 TI - [Clinical study of chronic myelogenous leukemia]. PMID- 8245627 TI - [Autoimmune thyroiditis]. PMID- 8245628 TI - [clinical study of heart enlargement]. PMID- 8245629 TI - [Autoimmune liver diseases]. PMID- 8245630 TI - [Prevention and early diagnosis of disseminated intravascular coagulation]. PMID- 8245631 TI - [Immune mechanism of digestive systems]. PMID- 8245632 TI - [Immunologic study of chronic gastritis]. PMID- 8245633 TI - [Inflammatory bowel disease and immunology]. PMID- 8245634 TI - [Chronicity and immunology of viral hepatitis]. PMID- 8245635 TI - [Physiopathology of autoimmune liver diseases]. PMID- 8245636 TI - [Classification of amyloidosis]. PMID- 8245637 TI - [Amyloidosis of amyloid AA and AL protein]. PMID- 8245638 TI - [Transthyretin and familial amyloid polyneuropathies]. PMID- 8245639 TI - [Beta/A4-type amyloid protein in Alzheimer's disease and cerebral amyloid angiopathies]. PMID- 8245640 TI - [Beta2-microglobulin and hemodialysis]. PMID- 8245641 TI - [Islet amyloid polypeptide and diabetes mellitus]. PMID- 8245642 TI - [New development on the study of etiological pathology of gastritis, peptic ulcer and stomach cancers in relationship to Helicobacter pylori infection]. PMID- 8245643 TI - [Diagnosis and therapy of pulmonary mycoses]. PMID- 8245644 TI - [Changing pattern of Chlamydia respiratory tract infections--with special reference to infections in Japan]. PMID- 8245645 TI - [Changing pattern in rickettsia infections--with special reference to tsutsugamushi disease and spotted fever]. PMID- 8245646 TI - [Changes in genes of hepatitis B virus and liver diseases]. PMID- 8245647 TI - [Proliferation mechanism of hepatitis C virus and effectiveness of interferon therapy]. PMID- 8245648 TI - [Chronic EB virus infections and cytokines--with special reference to IL-2, interferon-gamma and BCRF1 gene products]. PMID- 8245649 TI - [Physiopathology of ulcerative colitis and the non-invasive therapy]. PMID- 8245650 TI - [Physiopathology and therapy of chronic pancreatitis]. PMID- 8245651 TI - [Diagnosis and therapy of ischemic heart diseases]. PMID- 8245652 TI - [Treatment of diabetes mellitus]. PMID- 8245653 TI - [DNA diagnosis of hemophilia]. PMID- 8245654 TI - [Diagnosis of dementia--with special reference to image diagnosis and biological markers]. PMID- 8245655 TI - [Internal diseases and serum enzymes]. PMID- 8245656 TI - [Laparoscopic cholecystectomy]. PMID- 8245657 TI - [Intravascular ultrasonography]. PMID- 8245658 TI - [Visceral fat obesity]. PMID- 8245659 TI - [Etiology of nephrotic syndrome]. PMID- 8245660 TI - [Clinical study of chronic pulmonary thromboembolism]. PMID- 8245661 TI - [Chronic fatigue syndrome]. PMID- 8245662 TI - [Adult-onset Still's disease]. PMID- 8245663 TI - [Infectious diseases transmitted from foreign countries]. PMID- 8245664 TI - The differential diagnosis and comorbidity of hidradenitis suppurativa and perianal Crohn's disease. AB - Over the last 8 years, 61 patients with hidradenitis suppurativa (HS) have been treated at this institution. Twenty-four have also had a diagnosis of Crohn's disease (38%). This dual pathology is examined in detail in this retrospective review. There were 11 males and 13 females with a mean age of 39 years (range 18 to 75 years). The Crohn's disease was ileal in 1 patient, ileocolic in 4, and affected the large bowel only in 19. The diagnosis of Crohn's disease predated that of HS by an average of 3.5 years. At the time of review, 22 patients had a stoma, 23 had undergone laparotomy and 17 had lost their rectum. Hidradenitis suppurativa occurred in the perineal or perianal area in all patients but involved other sites in 20 cases. Skin grafting had been done in 9 and local procedures in 19 patients. Granulomas were found in excised skin in 6 cases but this finding was not associated with a poor outcome. At a mean follow-up of 3.2 years from the most recent surgery for HS (range 1 to 11 years) 11 were asymptomatic for HS, 11 had symptoms and no follow up was available in 2. These data show that HS may coexist with Crohn's proctocolitis, complicating the diagnosis and management of patients in whom it occurs. An increased appreciation of the possibility is recommended. PMID- 8245665 TI - Decrease in collagenous proteins and mechanical strength of distal colon after diverting colostomy in rats. AB - The effects of a diverting colostomy on weight, collagen content, and biomechanical strength of the left colon was investigated in rats. Female rats received either a diverting colostomy or a sham-operation and were killed randomly 1, 2, 4, or 12 weeks after surgery. The dry weight and hydroxyproline content of excluded colon decreased by 37% (P < 0.001, confidence limits (CL) = 21-53%) and 25% (P < 0.001, CL = 18-31%), respectively, after 4 weeks, and by 52% (P < 0.001, CL = 38-66%) and 40% (P < 0.001, CL = 27-53%), respectively, after 12 weeks compared to sham-operated rats. The breaking strength of the defunctioned colon decreased by 21% (P < 0.05, CL = 2-40%) after 4 weeks and by 25% after 12 weeks (P < 0.05, CL = 4-46%). The tensile strength of defunctioned colon (load per unit collagen) was reduced by 19% after 4 weeks (P < 0.05, CL = 6-32%). Proximal to the colostomy site we found no differences in the hydroxyproline content or biomechanical strength. In conclusion, defunctioning of the left colon in rats leads to deterioration in the biomechanical properties caused by a lesser content and a decreased quality in the collagen of the left colon. PMID- 8245666 TI - Anal manometry in patients with fissure-in-ano before and after internal sphincterotomy. AB - The motility pattern of the internal anal sphincter was estimated manometrically in 42 patients with fissure-in-ano before and after left lateral internal sphincterotomy (LAS). Resting anal pressure and anal pressure during straining were significantly higher in patients than in controls (132 +/- 21 SD cmH2O vs 81 +/- 14 SD cmH2O P < 0.0002 and 46 +/- 16 SD cmH2O vs 13 +/- 4 SD cmH2O P < 0.0005), but were normal after LAS. Slow waves were more common in fissure patients (86 +/- 6 SD% of total recording time vs 68 +/- 11 SD% of total recording time, P < 0.0002), but also became normal after successful treatment. The presence of ultra slow waves was also more common in fissure patients (P < 0.0001), and although it was significantly reduced postoperatively (P < 0.0001), it did not return to normal. Sampling was less frequent in fissure patients (P < 0.0001) and improved significantly after successful treatment (P < 0.0002). Rectal distension produced significantly less reduction in anal pressure in fissure patients as compared to controls (P < 0.01), but successful treatment returned the response to normal. There were 2 patients with anal fissure who did not heal after left LAS. Those patients and a further 5 patients with non healed fissures after left LAS showed the same pathological manometric features as before surgery. Their fissures were successfully treated by additional right lateral internal sphincterotomy. In conclusion, increased internal sphincter activity is probably an aetiological factor in fissure-in-ano, while successful LAS improves anal sphincter function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245667 TI - Effect of perioperative blood transfusions on survival of patients after radical surgery for colorectal cancer. AB - The effect of perioperative blood transfusion on the survival of patients with colorectal cancer was evaluated in 128 patients undergoing curative surgery between 1980 and 1988. The following clinical and histopathological variables were also studied: age, sex, duration of symptoms, presence of intestinal obstruction, tumour site, extent of spread through the bowel wall, lymph node involvement, Dukes' stage, grade of differentiation, venous invasion and type of surgical procedure performed. The need for perioperative blood transfusion was unrelated to the stage of disease. In the transfused patients (n = 73) the 5-year recurrence-free survival, calculated by the Kaplan-Meyer technique, was 37% and in the non-transfused (n = 55) was 60% (P = 0.0027, Mantel-Cox). Similar differences were found in the comparison of the groups with (n = 68) and without (n = 60) transfusions on the day of operation. The deleterious effect of transfusion was evident in patients who received only one unit of blood (n = 19)- these had a 5-year survival rate of 45% compared those who had more than one unit of blood (n = 54) (5-year survival rate 35%) (P = 0.0062). With a multivariate analysis, using a Cox proportional hazard model, taking into account all the variables studied, a significant and independent effect on survival was found for lymph node involvement (beta coefficient = 3.97), blood transfusion (beta coefficient = 2.16) and extent of bowel wall spread (beta coefficient = 1.75).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245668 TI - Our approach to the management of congenital presacral tumors in adults. AB - This study reviews the recent overall experience in one colorectal surgery department with congenital presacral tumors in adults. 24 patients greater than 21 years of age, who underwent curative resection between January 1980 and August 1992, were analyzed retrospectively. The growths were divided into two broad categories: developmental cysts and chordomas. The most common presenting symptom was pain (19/24). A preoperative evaluation regimen is outlined in the study and includes use of CT scanning, MRI imaging, and possibly the use of endoluminal ultrasound to document the relationship of presacral tumors to pelvic viscera. There were 20 developmental cysts and 4 chordomas treated in this series. 15 of 19 developmental cysts were excised by a posterior approach alone, 2 were excised by an anterior approach alone, and 3 were treated by a combined approach. Trans sacral excision was carried out in 4 patients with developmental cysts. One chordoma was resected posteriorly and the other 3 through a combined anterior and posterior approach. Three recurrences were diagnosed after excision of developmental cysts at 8, 18, and 41 months postoperatively. Recurrence occurred in 3 of 4 chordoma patients after 25, 32, and 55 months. Reexcision was carried out in all patients. None of the developmental cyst cases developed a second recurrence but 2 of the 3 chordoma patients have recurred, but have undergone local irradiation, which has controlled their disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245669 TI - Quality control in colonoscopy: a Singapore perspective. AB - For quality control purposes, a computer based auditing system for colonoscopy has been developed in the Department of Colorectal Surgery, Singapore General Hospital, Singapore. The first year's experience was reviewed to determine both workload patterns and the individual operators' performances. Colonoscopic completion rates, both as raw data and cusum transformations, were used as indices of quality control. The mean colonoscopic completion rate for the Unit was 97.0% and, when temporal trends were analysed by cusum, all operators consistently met or exceeded the minimum target completion rate of 90%. PMID- 8245670 TI - Pattern of recurrence in rectal carcinoma: implications for the design of (future) studies on adjuvant therapy. AB - 255 patients who underwent potentially curative surgical treatment for rectal carcinoma at the Cantonal Hospital Aarau from 1981 to 1989 have been followed up and the results analysed for overall survival and the timing and occurrence of local and distant recurrence in relation to the initial tumor stage. The patients had received standard surgical treatment for rectal carcinoma with the exception of 28 patients who had been randomized to the treatment arm of SAKK protocol 40/81 and were given perioperative adjuvant portal chemotherapy. 10 patients with large fixed tumors had preoperative radiation therapy. In UICC stage I most patients were cured by surgery alone, local recurrence as rare, and distant metastases appear in about 10% of pT2. cases. In stage II about one fifth of the patients developed a recurrence with one third of them having only local recurrence, and two thirds having either combined local and distant recurrence or distant recurrence alone. The small subgroup of pT4 pN0 had a very poor prognosis with early occurrence of distant metastases. In stage III the actuarial five-year survival was 25%, 60% of stage III cases had pN1 nodal disease. No patient with pN2-3 disease was alive after five years follow-up. The overall risk of distant metastases in stage III was 64%. Local recurrence rate was highly dependent on the pT-stage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245671 TI - Parenteral single dose ceftriaxone with tinidatsole versus aminoglycoside with tinidatsole in colorectal surgery: a prospective single-blind randomized multicentre study. AB - A single dose preoperative antibiotic prophylaxis of ceftriaxone 2 g and tinidatsole 500 mg was compared with netilmycin (150 mg) or tobramycin (80 mg) in patients undergoing colorectal surgery by a prospective randomised single-blind trial. There were 628 patients from seven Finnish hospitals. The wound infection rate of 3% in the ceftriaxone group was significantly lower than 12% of the aminoglycoside group. Wound infection was related to the amount of perioperative blood loss, duration of the operation, type of operation and drainage in the aminoglycoside group. In the ceftriaxone group, wound infection was related to the patient's weight only. PMID- 8245672 TI - CCK-8 and gastrin plasma levels in cholecystectomized and colorectal cancer patients. AB - The elevated incidence of large bowel carcinoma after cholecystectomy has long been controversial. The pathomechanism of this entity, however, is still unclear. Many authors have demonstrated a correlation between cholecystokinin (CCK) and gastrin levels and the occurrence of colorectal cancer. As yet, no clear data are available on the potential impact of cholecystectomy on CCK level alterations. Moreover, no reports have yet been published on CCK receptors. We have investigated the role of CCK-8 and gastrin plasma levels in patients with prior cholecystectomy and CCK receptor levels in patients with colorectal cancer. 125 patients entered a prospective study. Of these, 45 served as controls. 40 patients had prior cholecystectomy, 5 patients underwent cholecystectomy during the ongoing trial. 35 patients had a colorectal cancer, 5 of these had prior cholecystectomy. No patient had elevated CCK-8 plasma levels. Gastrin levels were slightly elevated in 2 patients. There was no correlation between large bowel carcinoma and CCK-8 and gastrin levels. Elevated CCK-8 levels following cholecystectomy occur neither immediately after surgery nor on a long-term basis. Immunohistochemical studies in patients with colorectal cancer showed no CCK receptors in the normal colonic or tumor tissue. These findings are contrary to gastrin receptor data. PMID- 8245673 TI - Operative mortality in carcinoma of the rectum. Results of the German Multicentre Study. AB - An analysis of operative mortality for carcinoma of the rectum in the German Study Group Colo-Rectal Carcinoma (SGCRC) comprising 1115 operated patients, found a total operative mortality rate of 4.3% and a 30-day-mortality rate of 2.8%. Advanced age, severe concurrent disease, anastomotic leakage in anterior resections and institution were found to be factors with independent significant influence. In anterior resections, especially low anterior resections, with protective enterostomy, the lower rates of anastomotic leakage and mortality emphasize the advantages of protective enterostomy. PMID- 8245674 TI - Nerve regeneration across colorectal anastomoses after low anterior resection in a canine model. AB - The disappearance and subsequent return of the recto-anal inhibitory reflex following low anterior resection was investigated using manometric and histological studies. Ten female greyhound dogs were randomised into 2 groups. All underwent low anterior resection. Five had stapled EEA anastomosis of the rectum and 5 were handsewn. The recto-anal inhibitory reflex was measured before and after surgery using a microtransducer tipped catheter. Six months postoperatively the recto-anal inhibitory response was again measured and the animals sacrificed. The colorectal anastomoses were examined histologically using light microscopy to determine the pattern of innervation at the anastomotic site. Manometric studies showed the recto-anal inhibitory reflex present in all cases pre-operatively, in only 1 case on the tenth postoperative day and was present in eight cases after 6 months. Histological examination of longitudinal sections across the anastomoses showed clear evidence of regenerating nerve trunks at the anastomotic site in both stapled and handsewn groups. We conclude that the return of the recto-anal inhibitory reflex is associated with regeneration of intramural autonomic nerves across the anastomotic scar. PMID- 8245675 TI - Level of anastomosis and anorectal manometry in predicting function following anterior resection for adenocarcinoma. AB - The clinical and physiological consequences of sphincter preservation after resection of rectal carcinoma at various levels were evaluated. Thirty-two patients (mean age 59.2 years; range 31-79 years) who had undergone curative surgery at least one year previously and were not given adjuvant therapy, were studied with 19 normal controls. Three blinded observers independently assessed clinical sphincter function, level of anastomosis (ANL) and anorectal manometry. 85.7% had an increase in stool frequency but their normal lifestyles were unaffected. The mean stool frequency was 3.1 (range 2 to 8) times/day. The anal maximal basal and maximum squeeze pressures, rectal volume of first sensation (VIS) and urge, maximum tolerable volume and compliance were all significantly impaired (P < 0.05). Patients with poorer function had significantly lower ANLs (P < 0.05). On multiple regression analysis, the ANL and the VIS significantly predicted stool frequency (P < 0.05). ANL below 6 cm was significantly associated with impaired frequency of stools (P < 0.05). This may be a subgroup of patients whose residual rectal reservoir function is compromised and may theoretically benefit from colonic pouches. PMID- 8245676 TI - Early post-operative 5-fluorouracil does not affect the healing of experimental intestinal anastomoses. AB - 5-Fluorouracil (5-FU) remains the mainstay for systemic treatment of colorectal cancer. In view of the increasing interest in peri-operative administration of antineoplastic agents, we have investigated the effects of early postoperative 5 FU on the healing of intestinal anastomoses in the rat. Animals underwent resection and anastomosis of both ileum and colon and 5-FU (20 mg/kg body weight) was given, either intravenously or intraperitoneally, on the day of surgery and the two subsequent days. Healing was assessed three and seven days after operation. Administration of 5-FU led to a reduced white blood cell count. However, anastomotic strength was not significantly reduced at either time point and anastomotic hydroxyproline content was not significantly affected. We suggest that limited use of 5-FU during or immediately after operation does not necessarily affect early anastomotic healing in the intestine. PMID- 8245677 TI - Successful local repair of paracolostomy hernia with a newly developed prosthetic device. PMID- 8245678 TI - Proopiomelanocortin in skin: new possibilities for regulation of skin physiology. PMID- 8245679 TI - Endothelin: localization in pheochromocytoma. PMID- 8245680 TI - Cortisol and hypertension: what was not so apparent about "apparent mineralocorticoid excess". PMID- 8245681 TI - IGF-I and the kidney. PMID- 8245682 TI - Peritoneal function and peritonitis in patients on peritoneal dialysis. PMID- 8245683 TI - Monocyte-endothelial interactions: insights and questions. PMID- 8245684 TI - Drug and environmental effects on the induction of autoimmunity. PMID- 8245685 TI - Detection of proopiomelanocortin-derived antigens in normal and pathologic human skin. AB - We investigated the presence of proopiomelanocortin (POMC) products in sections of skin from normal subjects and patients with neoplastic and non-neoplastic cutaneous disorders. Antibodies specific against adrenocorticotropin, beta melanotropin, and beta-endorphin were used for detection and characterization of cell types bearing POMC peptides. POMC products were not observed in sections of normal skin from the corporal (non-scalp) areas (six cases), whereas the hair follicles of scalp skin exhibited positive immunostains that were readily apparent (four cases). POMC products were frequently detected in corporal skin affected by diseases (13 of 26 cases), for example, psoriatic keratinocytes, the inflammatory infiltrate in scarring alopecia, nevocytes, the epithelial cell nests of basal cell carcinoma, and melanoma cells. Further tests were performed in keloids, a primary reactive skin disorder, to evaluate whether POMC accumulation represented a disease-related phenomenon or an expression of normal cutaneous reactivity. POMC products were consistently detected (10 of 11 cases) in the keratinocytes and mononuclear cells at keloid lesions. Thus these observations indicate that POMC products may accumulate locally in lesional skin representing, presumably, a novel cutaneous response to injury. The broad spectrum of POMC products detected suggests that these arise from production in situ (expression of the POMC gene itself) by human skin. PMID- 8245686 TI - The effect of carbenoxolone on the peripheral metabolism of cortisol in human patients. AB - Carbenoxolone in human patients induces a state that is similar to that seen in the syndrome of apparent mineralocorticoid excess. The mechanism in both the drug induced and the naturally occurring disorder is thought to be the inhibition of a normal mechanism for preventing access of cortisol to the mineralocorticoid receptor, namely 11 beta-hydroxy dehydrogenation. We took the opportunity to study the effect of carbenoxolone on the peripheral metabolism of cortisol in the course of evaluating the drug's therapeutic effectiveness in pseudohypoaldosteronism. Carbenoxolone, at a dose that induces mineralocorticoid effects in patients with normally responsive mineralocorticoid receptor systems, did not lead to significant changes in the urinary cortisol: cortisone tetrahydrometabolite ratio. There was, however, a marked inhibition of ring A reduction of both cortisol and cortisone to tetrahydro metabolites. Urinary cortisol level was not significantly changed, but urinary cortisone level was decreased and the cortisol:cortisone ratio markedly increased. We conclude that the urinary cortisol:cortisone tetrahydrometabolite ratio is not necessarily a valid measure of effective inhibition of 11 beta-hydroxy dehydrogenation. A better measure of the inhibitory effect of carbenoxolone on 11 beta-hydroxy dehydrogenation is the urinary free cortisol:cortisone ratio. PMID- 8245687 TI - Effect of dietary protein intake on renal growth: possible role of insulin-like growth factor-I. AB - Insulin-like growth factor-I (IGF-I) has been implicated as a possible mediator of renal hypertrophy after uninephrectomy and diabetes mellitus. Because renal hypertrophy is also a consequence of high protein intake, we studied the effect of varying concentrations of dietary protein on circulating levels and renal tissue content of IGF-I. Male Sprague-Dawley rats were fed isocaloric diets containing high (50%, HP), normal (20%, NP) or low (6%, LP) dietary protein for up to 14 days before they were killed. As expected, renal size (dry kidney weight) was greater in HP-fed rats and smaller in LP-fed rats when compared with NP-fed animals (HP, 1415 +/- 26 mg [p < 0.01 vs NP]; NP, 1148 +/- 27 mg; LP, 838 +/- 16 mg [p < 0.01 vs NP]), and most of the relative changes in kidney size occurred during the first week of ingestion of the experimental diet. Renal hypertrophy in the HP-fed animals was accompanied at day 3 by a significant rise in kidney tissue IGF-I that remained elevated at day 7 but had fallen to baseline values by day 14. The rise in renal IGF-I content in the HP-fed rat was accompanied by increases in circulating IGF-I on day 3 only. Both circulating and renal tissue IGF-I levels were suppressed in the LP-fed animals at 3, 7, and 14 days. These data confirm that varying dietary protein intake has profound effects on both circulating and renal IGF-I levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245688 TI - Relationship of TNF-alpha, interleukin-6, and prostaglandins to peritoneal permeability for macromolecules during longitudinal follow-up of peritonitis in continuous ambulatory peritoneal dialysis. AB - Infectious reactions are known to comprise changes in vasopermeability and inflammatory mediators. We used peritonitis that complicated continuous ambulatory peritoneal dialysis (CAPD) as an in vivo inflammation model to study the time courses of peritoneal permeability characteristics and mediators in dialysate. Sixteen episodes of peritonitis were prospectively followed on eight consecutive days from the onset of the infection and once after recovery (control). Dialysate night dwells were examined for marker proteins of peritoneal permeability, cytokines (tumor necrosis factor-alpha [TNF-alpha], interleukin-6 [IL-6] and prostaglandins (PGE2, 6-keto-PGF1 alpha, and thromboxane B2 [TxB2]). The clearance of beta 2-microglobulin was used as indicator of the effective peritoneal surface area. The intrinsic permeability was characterized functionally by the peritoneal restriction coefficient. All protein clearances were increased during the acute phase of peritonitis and subsequently decreased to control. Likewise, the intrinsic peritoneal permeability was elevated, as demonstrated by a decrease of the peritoneal restriction coefficient. Peritoneal appearance rates of TNF-alpha, IL-6, and prostaglandins were also increased during the acute phase. Peak values were reached on day 1. The largest increase was observed for IL-6 (median 854-fold), followed by TNF-alpha (35-fold). The vasodilating PGE2 and 6-keto-PGF1 alpha were increased 12-fold and the vasoconstricting TxB2 was increased threefold. Evidence was obtained for local intraperitoneal synthesis of IL-6 and prostaglandins. TNF-alpha production appeared to be present only during the early acute inflammatory response. Analysis of variance for repeated measurements revealed that changes in the clearance of beta 2-microglobulin were related to those in IL-6 and marginally also to TNF-alpha in dialysate. Changes in the peritoneal restriction coefficient were also related to IL-6, but were more closely related to alterations in dialysate PGE2. These findings suggest that TNF-alpha, IL-6, and PGE2 are involved in the changes in permeability characteristics during CAPD-related peritonitis. PMID- 8245689 TI - The effect of chemical modification of basic amino acid residues on the activation and amidolytic activity of Hageman factor (factor XII). AB - Modification of arginyl residues of Hageman factor by phenylglyoxal hydrate inhibits activation of this clotting factor in a plasma-free system, that is, in the absence of the other constituents of the contact activation system. Activation is also inhibited by alteration of the other two basic amino acid residues present, lysine and histidine. Chemical modification of histidine and arginine residues does not inhibit the amidolytic activity of activated Hageman factor. In contrast, modification of amino group(s) in N-terminal and lysine residues inhibits activated Hageman factor. Thus, basic amino acid residues essential to the activation or activity of Hageman factor appear to be variably accessible to chemical modification. PMID- 8245690 TI - Modulatory effect of arachidonate 5-lipoxygenation on glomerular cell proliferation in nephrotoxic serum nephritis. AB - The modulatory effect of arachidonate 5-lipoxygenation on glomerular cell growth was assessed in rat nephrotoxic serum nephritis (NSN). After a single intravenous injection of anti-glomerular basement membrane immune serum, significant increments in glomerular proliferative activity (GPA)--assessed by tritiated thymidine incorporation in short-term glomerular cultures--occurred and were associated with enhanced glomerular cell proliferation, assessed in cortical sections by staining for the presence of proliferating cell nuclear antigen (PCNA) positive cells in glomeruli. Leukocyte depletion induced by x irradiation ameliorated the enhanced GPA and reduced PCNA (+) cell counts. The same effect was observed after treatment of rats with the arachidonate 5-lipoxygenase inhibitor MK886. These observations indicate that in NSN, leukocytes infiltrating glomeruli, and leukocyte-derived arachidonate 5-lipoxygenation eicosanoids promote glomerular cell proliferation. PMID- 8245691 TI - Vitamin K-dependent clotting factors are elevated in young adults who have close relatives with ischemic heart disease. AB - An increase in factor VII was found to be a risk factor for ischemic heart disease. The present study was designed to test the hypothesis that this increase in factor VII is part of a general increase in vitamin K-dependent clotting factors. Initially, a prospective analysis of factor VII antigen and prothrombin activity was performed in two groups of young subjects without symptoms who differed in their risk of ischemic heart disease based on a history (or lack thereof) of premature heart disease in a first-degree relative. A statistically significant increase in prothrombin activity and factor VII antigen was found in the high-risk group of subjects when compared with the low-risk group. In a second series of subjects, factor IX and X activity assays were also performed, and all four of the vitamin K-dependent clotting factors were found to be significantly higher in high-risk subjects when compared with low-risk subjects. A second goal of the study was to explore whether correlations between factor VII and cholesterol and triglycerides might be due to binding of factor VII with apolipoprotein B. Although a significant correlation of factor VII antigen with apolipoprotein B (rho = 0.523, p < 0.025) was found in our high-risk group of subjects, the correlation between factor VII and triglycerides (rho = 0.641, p < 0.005) was even stronger statistically, suggesting a probable interaction of factor VII with very-low-density lipoproteins in vivo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245692 TI - Altered pharmacokinetic properties of a lipophilically derivatized low-molecular weight heparin in rats. AB - A new generation of lipophilic heparins has been developed that show longer lasting inhibitory effects on the coagulation system. We have studied the radiopharmacokinetics of a derivatized low-molecular-weight heparin (LMWH) with a residualizing lipophilic tyramine-deoxysorbitol label in comparison with conventional LMWH after intravenous application into Wistar rats. Whole body scintigraphy and analysis of the blood and organ distribution of different tracer preparations revealed that the lipophilically derivatized LMWH substance was predominantly trapped in the liver RES by a scavenger receptor-mediated mechanism. After the saturable uptake mechanism was blocked by maleylated bovine serum albumin, 41.4% of the lipophilic LMWH tracer circulated in blood, as compared with 18.4% of the control and 1% of conventional LMWH. The same results were attained by a competition experiment with an excess of unfractionated heparin. Urinary excretion of the lipophilic tracer among the rats in this competition experiment was considerably lower (13.7%) as compared with conventional LMWH (53.0%). Experiments with lipophilic LMWH tracer bound nonspecifically to rat serum albumin confirmed that the prolonged half-life might in part be due to an increased affinity for albumin. About 59% of the activity of the lipophilic tracer bound to albumin was found in the liver reticuloendothelial system, and only 3.3% were excreted to urine 3 hours after injection. Further studies are necessary to evaluate the accumulation rates and the metabolic fate of lipophilically derivatized heparins in the case of an impeded reticuloendothelial system uptake before attempts are made to therapeutically apply these compounds. PMID- 8245693 TI - Pregnancy decreases the threshold for cocaine-induced convulsions in the rat. AB - The objective of this study was to test our hypothesis that pregnancy modifies the central nervous and cardiovascular toxicity of cocaine. Ten chronically catheterized term pregnant rats and 13 chronically catheterized nonpregnant female rats were infused with cocaine (2 mg/kg/min) intravenously to observe the sequential toxic manifestation of cocaine from mild central nervous stimulation (hyper-locomotor activities) to fatal cardiovascular collapse. Arterial blood samples were withdrawn at the onset of major toxic signs or symptoms--namely convulsion, hypotension, and circulatory collapse--for determination of cocaine concentrations and plasma cholinesterase activity. The dosage and plasma concentrations of cocaine associated with the onset of convulsions and cardiovascular depression were significantly lower in pregnant rats when compared with the nonpregnant animals. The mean time required to develop convulsions in the pregnant rat was significantly shorter (21 minutes) than that in the nonpregnant animal (33 minutes). However, once convulsive activity had developed, the time interval to achieve circulatory collapse was similar in both groups. Although the baseline plasma cholinesterase activity was higher in the pregnant rats than in the nonpregnant ones, the values in the samples obtained from the pregnant group at the onset of circulatory collapse were similar to the baseline values for the nonpregnant group. These findings suggest that a higher enzyme activity does not protect the development of toxic manifestations in the pregnant rat as compared to the nonpregnant animal when cocaine was administered at the same infusion rate. PMID- 8245694 TI - Gender and reading disability. PMID- 8245695 TI - Learning disabilities in Colombia, South America. PMID- 8245696 TI - Dysrationalia: a new specific learning disability. AB - The concept of selective deficit is the foundation of most conceptual definitions of learning disability. Such definitions have tended to implicate the construct of intelligence in the conceptualization of learning disability and have led to the use of IQ test scores to operationalize the notion of aptitude-achievement discrepancy. The learning disabilities field is only beginning to grapple with the implications of its reliance on the concept of psychometrically defined intelligence. For example, discrepancy-based definitions of learning disabilities guarantee that such disabilities will become more or less prevalent depending on the comprehensiveness of the set of skills assessed on IQ tests. Unlike the vernacular concept of intelligence--which is quite broad--psychometric operationalizations reflect only a thin slice of the mental domain that might be considered cognitive. Thus, it is possible that we have not exhausted the potential set of discrepancy-based disabilities. As a demonstration proof, a new discrepancy-based disability category is proposed and defended in this paper. The disability is one that may force more careful consideration of the role that intelligence plays in conceptual and operational definitions of learning disabilities. PMID- 8245697 TI - Would you rather take orders from Kirk or Spock? The relation between rational thinking and intelligence. AB - This article considers the relation between rationality and intelligence. The article is divided into three main parts: In the first, I consider the concept of rationality and conclude that rational thinking is a part of intelligence, but not a major part. In the second, I consider Stanovich's concept of dysrationalia, and propose that an alternative concept, practical intelligence (or the lack thereof), may better accomplish the goals that Stanovich wishes to achieve with his concept. Finally, I draw some conclusions. PMID- 8245698 TI - How many learning disabilities are there? A commentary on Stanovich's "Dysrationalia: a new specific learning disability". PMID- 8245699 TI - Use of constant time delay in teaching multiplication facts to students with learning disabilities. AB - This study investigated the effectiveness and efficiency of a constant time-delay procedure in teaching multiplication facts to three male and two female elementary school students, aged 9 years 6 months to 12 years 5 months, identified as learning disabled. The five students were taught unknown facts using a 4-second constant time-delay procedure. Data were collected on generalization to a paper-and-pencil task, to a horizontal display orientation, and to the reverse fact, as well as incidentally acquired ability to solve missing-factor problems. Results indicated that the near-errorless learning procedure was effective and efficient in teaching multiplication facts to students with learning disabilities. Learning generalized to a paper-and-pencil task, to a different presentation orientation, and to the reverse fact. PMID- 8245700 TI - What do students with learning disabilities think when their general education teachers make adaptations? AB - The purpose of this study was to investigate the perceptions of mainstreamed students with learning disabilities (LD) regarding adaptations (e.g., altering tests, homework, assignments, instruction) made by general education teachers. Furthermore, their responses were compared with those of low achieving (LA) and average/high achieving (A/HA) classmates. One hundred seventy-nine students participated in this study: 60 mainstreamed students with LD, 59 low achieving students, and 60 average/high achieving students. Students were selected from 60 teachers' classrooms (20 elementary, 20 middle school, and 20 high school). Results from the elementary level indicate that students with LD differ from their LA and A/HA classmates in that the former indicate a stronger preference for opportunities to work in groups with different students, and prefer the teacher to make adaptations when they have difficulty learning. High school and middle school students from both the LA and A/HA groups, but not the LD group, preferred the teacher who made no adaptations in homework and textbooks. Discussion focuses on the role of students' perceptions in teacher decision making. PMID- 8245701 TI - External validity of the profile variability index for the K-ABC, Stanford-Binet, and WISC-R: another cul de-sac. AB - Clinicians often attach interpretive significance to high variability among subtest scores in IQ profiles of adults and children. In this study we evaluated the external validity of a relatively new measure of IQ subtest variability, the profile variability index (PVI). Within a sample of referred children, we administered the Kaufman Assessment Battery for Children, the Fourth Edition Stanford-Binet, and the Wechsler Intelligence Scale for Children-Revised. We calculated PVIs for each child from each test and correlated these values with achievement scores and indexes of discrepancy between actual and predicted scholastic achievement. Against these external criteria, PVI information from all three cognitive-ability batteries had essentially nil validity. We discuss implications of these findings for future research in this area. PMID- 8245702 TI - Enhanced hepatic clearance of intravenously administered sterically stabilized microspheres in zymosan-stimulated rats. AB - The blood clearance and organ deposition of sterically stabilized (poloxamine-908 coated) polystyrene microspheres of two different sizes (60 and 220 nm in diameter) were compared in control and zymosan-stimulated rats 3 h after intravenous administration. Poloxamine coating dramatically decreased the uptake of 60-nm microspheres by organs of the reticuloendothelial system and, concomitantly, kept microspheres in the blood. Large poloxamine-coated microspheres (220 nm) initially remained in the blood, but eventually a large fraction of these microspheres was filtered by the spleen. Daily administration of zymosan produced a marked increase in the intravascular clearance of the large, but not the small, poloxamine-coated microspheres. The enhanced intravascular clearance of large poloxamine-coated microspheres in zymosan treated rats was the result of hepatic sequestration. On the other hand, the splenic filtration of these microspheres was depressed by 225% below the control values, despite the dramatic increase in spleen size of zymosan-treated rats. Preincubation of large poloxamine-coated microspheres in serum derived from both the control and zymosan-treated animals suggested that the enhanced hepatic uptake of large sterically stabilized microspheres following zymosan stimulation was not the result of "specific opsonization" processes. Instead, the changes in the proliferative as well as the phagocytic response of Kupffer cells appeared to be responsible for these observations. The preferred hepatic uptake of large poloxamine-coated microspheres, as opposed to smaller particles, is suggested to be due to differences in surface characteristics and the properties of microspheres. These may include differences in polymer density and the surface conformation of the polyoxyethylene segments of the polymer in the biological environment and the way they interact with both plasma components and the macrophage surface. These observations could be of importance in the use of sterically stabilized drug carriers for delivery of therapeutic agents to sites other than the reticuloendothelial system in clinical conditions associated with globally or regionally enhanced reticuloendothelial activity. PMID- 8245703 TI - Effect of lipoxins and other eicosanoids on phagocytosis and intracellular calcium mobilisation in rainbow trout (Oncorhynchus mykiss) leukocytes. AB - Rainbow trout (Oncorhynchus mykiss) macrophages generated lipoxin (LX) A4, LXA5, leukotriene (LT) B4, LTB5 and 12-hydroxyeicosatetraenoic acid (12-HETE) during the phagocytosis of zymosan and Escherichia coli, but not of the yeast Saccharomyces cerevisiae. Prostaglandin (PG) E2 was also detected in supernatants from macrophages incubated with either zymosan or calcium ionophore A23187. LXA4 (10(-8)-10(-6) M) and LTB4 (10(-9)-10(-7) M) provoked rapid and transient dose dependent increases in intracellular calcium ([Ca]i) concentrations in leukocyte suspensions containing 40-60% macrophages. EC50 values were 14.9 and 1.2 nM, respectively. PGE2 and 12-HETE had no effect on [Ca]i at concentrations up to 30 microM. PGE2 and 12-HETE (10(-5)-10(-10) M) enhanced the in vitro phagocytosis of yeast test particles by trout macrophages, whereas LXA4 and LTB4 had no demonstrable effect on the responses of these cells at concentrations up to 10( 5) M. In conclusion, the processes involved in trout macrophage stimulation are complex but involve generation of both cyclooxygenase and lipoxygenase products. The increase in [Ca]i caused by LXA4 and LTB4 may form part of the chemotactic transduction mechanism that recruits granulocytes and macrophages to sites of inflammation. The effects of eicosanoids on phagocytosis appear to be independent of changes in [Ca]i. PMID- 8245704 TI - Granulocyte-macrophage colony-stimulating factor plays a role in the functional activity of mast cells. AB - A peptide homologous to a region of murine granulocyte-macrophage colony stimulating factor (mGM-CSF), P27-38, which was shown to be a GM-CSF antagonist, inhibited the function of serotonin release from murine mast cells. Peptide P27 38 inhibited immunoglobulin E (IgE)-mediated serotonin release in a dose dependent manner when induced by either specific antigen or anti-IgE antibody. In contrast, non-receptor-mediated release of serotonin by agents such as compound 48/80 or the calcium ionophore A23187 were not affected by the GM-CSF antagonist. Similar effects were observed with GM-CSF-neutralizing antibodies. The inhibitory effect of P27-38 and the neutralizing antibodies on serotonin release could be reversed by the addition of exogenous GM-CSF to the stimulated mast cells, indicating that the inhibitory activity was probably due to an effect on endogenously produced GM-CSF. These findings suggest that GM-CSF produced by stimulated mast cells is involved in the regulation of their activity in an autocrine manner. PMID- 8245705 TI - Expression of human perforin in a mouse cytotoxic T lymphocyte cell line: evidence for perturbation of granule-mediated cytotoxicity. AB - Expression of the pore-forming protein perforin is normally restricted to the cytolytic granules of cytotoxic T lymphocytes and natural killer cells. Perforin, which causes cell death by osmotic lysis, has the ability to form transmembrane channels in target cell membranes. This function makes perforin crucial in the granule-exocytosis model of T cell-mediated cytotoxicity. In the present study, variants of the mouse cytotoxic T lymphocyte cell line CTLL-R8 have been produced which express human perforin. A full-length cDNA clone (HP-10) encoding human perforin was inserted in the sense orientation into the expression plasmid pCMV5neo. The resultant construct, designated pCMV5neoHP-10, was used to transfect CTLL-R8 cells. Of eight G418-resistant clones studied, four clones expressed human perforin mRNA by Northern analysis and three of these clones also expressed human perforin protein by Western blotting. The expression of human perforin protein was associated with a pronounced (55-74%) and consistent reduction in the killing of three target cell lines, P815, YAC-1, and EL4, compared with parental CTLL-R8 cells. The reduction in target cell lysis could not be attributed to nonspecific effects of the transfection, as clones transfected with neo alone showed no reduction in killing in comparison with parental CTLL-R8 cells. Clones expressing human perforin showed very similar growth characteristics, surface phenotype, and N-alpha-benzyloxycarbonyl-l thiobenzyl-esterase release compared with untransfected CTLL-R8 cells. The mechanism of reduction of cytolysis is unclear but may involve competition by human perforin in the handling or packaging of endogenous granule constituents (including mouse perforin) or assembly of human perforin into mouse polyperforin channels in target cell membranes. The expression of human perforin in mouse cytotoxic T cells provides a potential model for studying how cytotoxic T cells process, package, utilize, and protect themselves against the perforin molecules they produce. PMID- 8245706 TI - Transient exposure of human eosinophils to the protein kinase C inhibitors CGP39 360, CGP41-251, and CGP44-800 leads to priming of the respiratory burst induced by opsonized particles. AB - We report that a transient incubation of human eosinophils with the protein kinase C (PKC) inhibitor CGP39-360 (staurosporine) or the more PKC-specific inhibitors CGP41-251 and CGP44-800 prior to activation of the respiratory burst with opsonized particles results in priming of this response. This priming effect was concentration dependent and occurred in the range in which the phorbol myristate acetate-induced respiratory burst was inhibited. CGP39-360 priming was minimally affected in Ca(2+)-depleted cells, indicating that an increase in [Ca2+]i is not important. Also, the binding of serum-treated zymosan (STZ) particles was strongly enhanced by the inhibitors. On the other hand, the release of platelet-activating factor (PAF) induced by opsonized particles was enhanced only by CGP39-360 and not by CGP41-251 and CGP44-800. Therefore, priming of the respiratory burst is not due to an aspecific enhancing effect of the inhibitors. These data indicate that different signal transduction routes are involved in priming of the STZ-induced respiratory burst and PAF release in human eosinophils. PMID- 8245707 TI - A recombinant amino terminal fragment of bactericidal/permeability-increasing protein inhibits the induction of leukocyte responses by LPS. AB - Bactericidal/permeability-increasing protein (BPI) is a major component of the granules of polymorphonuclear neutrophils (PMNs) and is involved in the killing of gram-negative bacteria. A 23-kd recombinant protein, corresponding to the NH2 terminal fragment of human BPI (rBPI23), has been shown to bind lipid A and antagonize some lipopolysaccharide (LPS)-mediated effects. In this study the ability of rBPI23 to prevent a wide range of cellular responses to LPS was investigated. In vitro assays were carried out using human blood to more closely approximate in vivo conditions. The release of proinflammatory cytokines [tumor necrosis factor (TNF), interleukin-1 beta (IL-1 beta), IL-6, IL-8], induced by E. coli O113 LPS, was markedly reduced by rBPI23 in a concentration-dependent fashion. The production of the anti-inflammatory protein IL-1ra (IL-1 receptor antagonist) was triggered by lower LPS concentrations than those necessary for the other cytokines. Furthermore, prevention of IL-1ra release required higher rBPI23 concentrations than for other cytokines. The LPS-induced production of oxygen-derived free radicals by phagocytic cells (resulting in chemiluminescence) was also prevented by rBPI23. The inhibition was specific for LPS because the activation of leukocytes by phorbol myristate acetate, zymosan, or TNF was unaffected by BPI. The ability of rBPI23 to antagonize specifically the effects of endotoxin in the complex environment of human blood along with its bactericidal activity suggests that rBPI23 may be a novel therapeutic agent in the treatment of gram-negative infections. PMID- 8245708 TI - Both mannose and beta-glucan receptors are involved in phagocytosis of unopsonized, heat-killed Saccharomyces cerevisiae by murine macrophages. AB - We studied the involvement of lectin-like receptors in phagocytosis of unopsonized heat-killed yeast (Saccharomyces cerevisiae) by murine macrophage like cell lines and murine peritoneal resident macrophages. For this purpose we used a technique that allowed us to discriminate ingested and adsorbed heat killed yeast. The internalization can be partly inhibited by soluble polyosides such as laminarin (beta-glucan) or alpha-mannan. However, when they were used together (0.4 mg/ml alpha-mannan and 0.4 mg/ml laminarin), almost complete inhibition of phagocytosis was obtained. These observations suggest that phagocytosis of unopsonized heat-killed yeast by murine macrophage-like cell lines as well as murine peritoneal resident macrophages is mediated by both mannose and beta-glucan receptors. The respective activity of these two types of receptors is a function of in vitro cell differentiation. To achieve maximal phagocytosis of unopsonized heat-killed yeast, coexpression of both mannose and beta-glucan receptors is required. PMID- 8245709 TI - A carboxyl-terminal tail peptide of neutrophil chemotactic receptor disrupts its physical complex with G protein. AB - The binding of G protein to the N-formyl peptide receptor of human neutrophils was investigated with site-specific synthetic peptides. Peptide CT336(322) (322RALTEDSTQTSDTAT336) from the carboxyl-terminal tail region of the receptor competed with the receptor for binding to bovine Gi protein. The peptide competition was assayed by dissociation of a GTP-sensitive, rapidly sedimenting (7S) form of receptor-G protein complex as analyzed by velocity sedimentation on linear sucrose density gradients. An IC50 of 590 microM was determined for CT336(322) peptide. A control peptide, with the reverse sequence, rCT322(336) (336TATDSTQTSDETLAR322), did not perturb the sedimentation of the reconstituted receptor-G protein complex up to the highest tested concentration, 3 mM. Other peptides tested, corresponding to central portions of the predicted intracellular loop regions CII140(127) (127VLHPVWTQNHRTVS140) and CIII239(227) (227KIHKQGLIKSSRP239) of the receptor, failed to dissociate the reconstituted receptor-G protein complex. Control peptides from the extracellular region EII184(170) (170KTGTVACTFNFSPWT184) and an unrelated sequence matching a portion of neutrophil cytochrome b, CYT306(296) (296KVVITKVVTHPFKTIE306), were also ineffective. Our results suggest that the cytoplasmic tail of the formyl chemotactic peptide receptor is involved in its coupling to the signal transducing G protein. PMID- 8245710 TI - Influence of suspension on the expression of protective immunological memory to murine Listeria monocytogenes infection. AB - Female BDF1 mice were immunized with virulent Listeria monocytogenes, which resulted in the generation of a long-lived state of protective immunological memory for this facultative intracellular bacterial pathogen. The influence of antiorthostatic suspension, a ground-based model employed to simulate certain aspects of weightlessness that occur during spaceflight, on the capacity of these mice to express memory immunity was evaluated. Memory-immune mice were suspended by the tail in the orthostatic or antiorthostatic position and were reinfected with a lethal dose of virulent L. monocytogenes. It was found that suspension did not influence the kinetics of bacterial growth in vivo when the rechallenge infection was started concurrently with the suspension. However, mice that were reinfected on day 2 or 4 of the suspension exhibited an enhanced capacity to eliminate the infection. Attenuation of this enhancing effect was observed when mice were infected on day 7 of the suspension. These results indicate that the stress of antiorthostatic suspension can influence the capacity of the murine host to express protective immunological memory to pathogenic bacteria. PMID- 8245711 TI - Histamine modulation of Ca2+ homeostasis in human neutrophils. AB - The influence of histamine on the basal intracellular free Ca2+ concentration ([Ca2+]i) and agonist-induced increases of [Ca2+]i was studied in Fura-2-loaded neutrophils. Histamine was unable to change the basal [Ca2+]i at concentrations (10(-6)-10(-4) M) that have been shown to cause a rapid increase in [Ca2+]i in a variety of cell types. Histamine, in contrast, was found to inhibit dose dependently the rise in [Ca2+]i induced by two neutrophil receptor agonists, N formylmethionyl-leucylphenylalanine (fMLP) and serum-opsonized zymosan particles. The histamine inhibition was shown to be specific for H2 receptor activation by blocking experiments with selective H1 and H2 receptor antagonists. In the absence of extracellular Ca2+, histamine failed to inhibit the agonist-induced rise in [Ca2+]i, indicating that histamine does not affect the release of Ca2+ from internal pools. Forskolin, which mimics the biochemical effects of H2 receptor activation by directly stimulating adenylate cyclase, also decreased the Ca2+ transients induced by receptor agonists. Similarly, 3-isobutyl-1 methylxanthine (IBMX), a phosphodiesterase inhibitor, reduced the Ca2+ response of activated neutrophils. These data suggest that in human neutrophils (1) no functional H1 receptors are present or alternatively H1 receptors are not coupled to cellular Ca2+ metabolism, and (2) H2 receptors modulate the receptor-triggered Ca2+ flux via the cAMP second messenger system. PMID- 8245712 TI - Docosahexanoic acid (22:6, n-3) but not eicosapentaenoic acid (20:5, n-3) can induce neutrophil-mediated injury of cultured endothelial cells: involvement of neutrophil elastase. AB - Previously published work has indicated that polyunsaturated fatty acids (PUFA) may enhance neutrophil-mediated damage to host tissues. We have found that endothelial detachment was significantly increased by neutrophils pretreated with docosahexaenoic (22:6, n-3) and arachidonic (20:4, n-6) acids at 10-40 microM but not by eicosapentaenoic acid (20:5, n-3). Endothelial cell lysis as measured by 51Cr release was unaffected. The extent of detachment was dependent on both fatty acid and neutrophil pretreatment concentrations. A specific leukocyte elastase inhibitor abrogated the increased detachment but catalase had no effect. Measurement of prostaglandin I2 synthesis as an alternative nonlytic assay of endothelial function indicated that 20:4 but not 20:5 was able to stimulate neutrophil-induced endothelial PGI2 synthesis. Although all three PUFA (3-33 microM) were found to stimulate release from neutrophil-specific granules, only 22:6 and 20:4 could stimulate release of the azurophilic granules containing elastase to any significant extent. Saturated fatty acids (20:0 and 22:0) and the methyl ester of 22:6 did not cause either neutrophil-mediated endothelial detachment or degranulation. We conclude that neutrophils pretreated with 22:6 or 20:4 but not 20:5 can decrease endothelial integrity through detachment involving neutrophil elastase. These findings may have important implications for the dietary use of fish oils rich in n-3 fatty acids. PMID- 8245713 TI - Dietary supplementation with n-3 fatty acids suppresses interleukin-2 production and mononuclear cell proliferation. AB - We studied the in vitro production of interleukin-2 in nine healthy volunteers who added 18 g/day of fish-oil concentrate rich in n-3 polyunsaturated fatty acids to their normal Western diet for a period of 6 weeks. Interleukin-2 synthesis from stimulated peripheral blood mononuclear cells was suppressed from 6.2 ng/ml at baseline to 2.2 ng/ml 10 weeks after the end of n-3 fatty acid supplementation (65% decrease; P = .04). At the same time phytohemagglutinin induced proliferation of mononuclear cells was suppressed by 70% from the presupplement level. Interleukin-2 production returned to the premedication level at the end of the studies. The results suggest that the effect of dietary n-3 fatty acids in some diseases may be mediated in part by decreased production of interleukin-2 and decreased mononuclear cell proliferation. PMID- 8245714 TI - Chemokines and serpentines: the molecular biology of chemokine receptors. AB - Chemokines are pro-inflammatory molecules with a diverse array of biological and biochemical functions. These molecules induce the migration of a number of leukocyte subsets including monocytes, neutrophils, and T-cells. The recent cloning of the IL-8, GRO, and MIP-1 alpha chemokine receptors revealed that these glycoproteins belong to the serpentine family of seven transmembrane G-protein coupled receptors. Other members of this family include the chemotactic receptors for fMLP and C5a, indicating that a common pathway for eliciting the directional migration of leukocytes is probably transduced via G proteins. Ligand binding to chemokine receptors is complex, featured by multiple chemokines binding to a single receptor and multiple receptors binding a specific ligand. Future directions in this field appear to be focused on the cloning of novel receptors and the identification of ligands for orphaned receptors. PMID- 8245715 TI - Involvement of inflammatory mediators in macrophage antitumor activity. AB - This review describes the potential role of macrophages in defense against cancer cells and the regulatory involvement of inflammatory mediators in this role. Interactions between macrophage-derived cytokines (tumor necrosis factor alpha, interleukin-1, IL-6) and their interrelationships with eicosanoids (mainly the cyclooxygenase product prostaglandin E2 and some lipoxygenase metabolites) represent a network that controls the expression of antitumor activity of macrophages either in a cell-to-cell contact system between the effector and the target tumor cell or as cell-free soluble products. Attention is given to the influence of tumor burden on production of cytokines and eicosanoids by macrophages and to the production of these mediators by tumor cells. Emphasis is placed on the roles of TNF-alpha and PGE2 in links between inflammatory and antitumor functions of macrophages. Finally, the perspectives and still existing problems in clinical implications of macrophage-derived cytokines are discussed in terms of a conceivable macrophage-directed immunotherapy of cancer. PMID- 8245716 TI - Role of liver in the maintenance of cholesterol and low density lipoprotein homeostasis in different animal species, including humans. PMID- 8245717 TI - Comparative binding of bile acids to serum lipoproteins and albumin. AB - Characteristics of the binding of lithocholic acid (LC), chenodeoxycholic acid (CDC), and cholic acid to human plasma proteins were studied. Affinity of the different plasma protein fractions for the bile acids studied decreased with increased polarity of the steroid nucleus of the bile acid. Binding of LC, CDC, and cholic acid to the lipoprotein-free, albumin-rich plasma fraction was characterized by two classes of binding sites with respective KDs of 2, 5, and 51 microM, and of 39, 2,387, and 5,575 microM, while corresponding Bmax values were similar for the different bile acids, at around 6 and 100 nmol/mg protein. Bile acid binding to the different lipoprotein fractions was characterized by a single population of binding sites, with a KD ranging from 47 to 66 microM for LC, 695 to 1010 microM for CDC, and 2,511 to 2,562 microM for cholic acid. Bmax values, at 416-913 nmol/mg protein, were similar among the different bile acids studied. Both glycine- and taurine-conjugated, as well as unconjugated LC competitively inhibited [24-14C]LC binding to low density (LDL) and to high density lipoproteins (HDL) to the same extent, while the more polar LC-3-sulfate, CDC, and cholic acid were increasingly less potent in displacing LC binding from the respective lipoproteins. Furthermore, all bile acids studied shared the same lipoprotein binding site. The lipoprotein fluorescence at 330-334 nm, following excitation at 280 nm, was diminished after incubation with LC, suggesting that the bile acid masks the tryptophan residues of the protein moiety. Finally, the initial rate of uptake of 1 microM LC, in isolated hamster hepatocytes, at around 0.045 nmol.sec-1.mg cell wt-1, was not affected by the protein carrier. However, for the same concentration of LC, bound to either LDL or HDL, LC binding resulted in 75-77% of the total [24-14C]LC nonspecifically bound to the hepatocyte, compared to 65% when bound to albumin, and 45% in the absence of protein. The studies show that, under conditions when the serum bile acid concentration exceeds the capacity of the high affinity class of albumin binding sites for bile acids, lipoproteins have similar or greater affinity to bind bile acids than does albumin. The ability of lipoproteins to increase the nonspecific association of lithocholic acid with liver cells may also facilitate bile acid association with extrahepatic tissues. As lipoproteins, in contrast to albumin, are targeted to most cells, they may play a major role in the transport of potentially toxic bile acids to peripheral cells. PMID- 8245718 TI - Bile acid excretion and cholesterol 7 alpha-hydroxylase expression in hypercholesterolemia-resistant rabbits. AB - We have developed a partially inbred substrain of New Zealand white rabbits (CRT/mlo) that are resistant to the hypercholesterolemia that accompanies cholesterol feeding to normal rabbits. The plasma cholesterol concentration of normal rabbits increases dramatically from about 30 mg/dl to > 300 mg/dl after they are fed a 0.1% cholesterol-enriched diet for 3-4 months. Cholesterol-fed CRT/mlo animals, however, maintain a cholesterol level of about 30 mg/dl during the entire cholesterol feeding period. In addition to the low plasma cholesterol level, measurements of cellular cholesterol indicate that the hepatic cholesterol content of the cholesterol-fed resistant rabbit remains markedly lower than it does in normal animals fed the same diet. The only mechanism for removal of significant quantities of cholesterol carbon from the body is via the fecal excretion of cholesterol, neutral sterol metabolites, and bile acids. In comparison to the basal, low-cholesterol diet, we observed that cholesterol-fed resistant rabbits had increased excretion of lithocholic acid, while excretion of this bile acid by cholesterol-fed normal rabbit remained similar to basal diet levels. Deoxycholic acid excretion, the other main bile acid excreted in the feces of rabbits, was decreased in response to cholesterol challenge in animals with either resistant or normal phenotypes, but the decrease was significantly less in the resistant rabbits. Thus, the resistant rabbits excreted relatively more lithocholic and deoxycholic acid than did the cholesterol-fed normal rabbit. The difference in bile acid excretion was also manifest by a higher than normal level of cholesterol 7 alpha-hydroxylase activity and cholesterol 7 alpha hydroxylase mRNA in the livers from resistant versus normal rabbits. As cholesterol 7 alpha-hydroxylase is the putative rate-limiting step of bile acid synthesis, we believe that the increased excretion of bile acids by resistant animals is due, at least in part, to increased levels of cholesterol 7 alpha hydroxylase expression. PMID- 8245719 TI - Association of plasma triglyceride concentration and LDL particle diameter, density, and chemical composition with premature coronary artery disease in men and women. AB - Low density lipoprotein (LDL) physical-chemical characteristics were studied as nontraditional risk factors of coronary artery disease (CAD) in a well characterized population of 98 men aged < or = 50 and 100 women aged < or = 60 who underwent elective diagnostic coronary arteriography. The average LDL diameter was determined by gradient gel electrophoresis, chemical composition (%w/w) was measured, and the density of the major LDL peak was determined by equilibrium density gradient ultracentrifugation. Logistic regression was used to examine the association of various LDL characteristics with CAD before and after adjustment for other covariates. Smaller, cholesterol-poor LDL particles were associated with CAD independently of traditional risk factors (age, sex, smoking, diabetes, LDL and HDL cholesterol concentrations), other than the plasma triglyceride concentration. These characteristics were generally more strongly associated with CAD when measured on the major LDL subfraction (defined as the density gradient ultracentrifugation fraction with the highest LDL concentration) than the average characteristics of the more heterogeneous parent LDL (d 1.019 1.063 g/ml). The associations with CAD among men and women were generally similar. These data show that a broad range of LDL characteristics are associated with CAD before, but not after, adjustment for the plasma triglyceride concentration. These data further indicate the importance of hypertriglyceridemia and LDL heterogeneity in premature CAD. PMID- 8245720 TI - Localization of a liver-specific enhancer in the apolipoprotein E/C-I/C-II gene locus. AB - The sequences necessary for liver-specific expression of the apolipoprotein (apo) E gene have been shown to reside 3' to the gene, within the apoE/C-I/C-II gene cluster, but have not been precisely characterized. Utilizing a transient transfection reporter gene assay based on the apoC-II promoter, we have localized a liver-specific enhancer to its approximate limit dimension of 154 base pairs. This enhancer directed liver-specific expression of an apoE gene construction in transgenic mice. A DNaseI protection assay revealed two footprints over an inverted repeat of a known transcriptionally active motif, TGACCT. DNaseI sensitive sites were present in three of six repeats of a motif (consensus GCAAACA) which has been postulated to represent the recognition sequence of a hepatic transcriptional activity, HNF-5. This region of DNA may function as a liver-specific enhancer for the entire apoE/C-I/C-II gene cluster. PMID- 8245721 TI - Metabolism of beta-muricholic acid in the hamster and prairie dog. AB - The metabolism of beta-muricholic acid was investigated in the prairie dog and the hamster. Intravenous infusion into bile fistula hamsters showed that beta muricholic acid was extracted by the liver and secreted into the bile (> 85% in 1 h). Hepatic extraction of this compound and cholic acid in the prairie dog was not as rapid as in the hamster. In the bile of the prairie dog, most (93%) of the administered beta-muricholic acid was present as the taurine conjugate. In the hamster, 28% of infused beta-muricholic acid was secreted in unconjugated form, 43% as the taurine conjugate, and 22% as the glycine conjugate. In both species, the administered compound underwent little biotransformation. After intraduodenal injection of [6 alpha-3H]-labeled beta-muricholic acid into bile fistula hamsters, the bile acid was rapidly secreted into the bile; more than 80% of the administered radioactivity was recovered in 3 h. In the prairie dog, biliary recovery after intraduodenal administration of either beta-muricholic acid (43% in 3 h) or cholic acid (22% in 3 h) was slower than in the hamster. After intragastric administration, more than 80% of beta-muricholic acid was recovered unchanged in feces of both animal species. PMID- 8245722 TI - Identification of disulfide-linked apolipoprotein species in human lipoproteins. AB - We wished to determine whether apolipoprotein C-IIToronto, a mutant form of apolipoprotein C-II that contains a C-terminal cysteine residue, exists as a monomeric species or as multiple disulfide-linked species in plasma lipoproteins. The plasma lipoproteins from a heterozygous carrier and two homozygous carriers of apoC-IIToronto were investigated. The mutant apolipoprotein was found in homodimeric form and as heterodimers with apolipoprotein A-II, apolipoprotein B 100, and apolipoprotein E. Of particular interest was the demonstration of the existence of the disulfide-linked species apolipoprotein B-100:A-II and B-100:C IIToronto in the very low density and low density lipoproteins in subjects who were carriers of apoC-IIToronto. We also observed that apoE3:C-IIToronto and apoE3:A-II dimers were present in the chylomicrons and very low density lipoproteins of these subjects. The observation of the existence of apolipoprotein B-100:A-II was extended to other hypercholesterolemic and hypertriglyceridemic subjects. The highest proportion of apolipoprotein B-100:A II was observed in the very low density lipoproteins of hypertriglyceridemic subjects. The concentration of this species was significantly higher in hyperlipidemic subjects than in normolipidemic controls. These results demonstrate that the molecular species of cysteine-containing apolipoproteins are complex and should be considered in studies of human lipoprotein composition and function. PMID- 8245723 TI - Cholesteryl esters in lymph chylomicrons: contribution from high density lipoprotein transferred from plasma into intestinal lymph. AB - Most of the cholesterol in intestinal chyle and chylomicrons is derived from plasma. Our aim was to determine how much plasma low density (LDL) and high density (HDL) lipoproteins contribute to the cholesterol in chyle and chylomicrons, and to examine how plasma cholesterol becomes associated with lymph chylomicrons. Intravenous injection of radioiodinated plasma lipoproteins into two chyluric patients showed that 82% of the HDL plasma pool transferred daily to intestinal chyle, corresponding to 58% of lymph cholesterol; LDL contributed 18% of its plasma pool, corresponding to 18% of lymph cholesterol. When plasma HDL radiolabeled in both the protein and cholesteryl ester moieties was injected, the isotope ratios of plasma HDL and lymph lipoproteins were identical; 85% of the HDL cholesteryl esters transferred to triglyceride-rich lipoproteins, while the apolipoproteins remained largely (70%) in the higher density lipoproteins of the chyle. Incubations of similarly labeled plasma HDL showed preferential transfer of cholesteryl esters to artificial chylomicrons mediated by a factor present in lipoprotein-free plasma. Thus, a sizable portion of plasma HDL enters intestinal lymphatics probably as intact HDL, and then transfers part of their cholesteryl esters to chylomicrons, possibly mediated by transfer proteins. Reverse cholesterol transport may therefore include an extravascular loop via lymph chylomicrons and chylomicron remnants to the liver. PMID- 8245724 TI - Effect of phosphodiesterase inhibition with amrinone or theophylline on lipolysis and blood flow in human adipose tissue in vivo as measured with microdialysis. AB - Phosphodiesterase III (cyclic GMP-inhibited, particulate, low Km) is believed to play a dominant role in the cyclic AMP breakdown and lipolysis regulation in fat cells. Its importance for lipolysis activity was investigated in situ in humans by comparing the effects of a selective (amrinone) and a nonselective (theophylline) inhibitor of the enzyme. Abdominal subcutaneous adipose tissue of healthy nonobese humans was microdialyzed with solvents containing one or both of these agents, and glycerol (lipolysis index) or the escape of ethanol from the dialysis solvent (blood flow index) was continuously monitored in the perfusate. Both agents caused a dose-dependent and sustained increase in the glycerol level in the perfusate for at least 2.5 h. Although amrinone was 5000 times more potent than theophylline on a molar basis, its maximum activity was only 35% as compared to the maximum activity of theophylline. Half-maximum lipolytic effect of the two drugs occurred at about 0.1 mumol/l and 1 mmol/l, respectively (P < 0.001). At maximum effective concentrations, amrinone stimulated lipolysis by about 63% and theophylline by about 200% (P < 0.01). At these concentrations amrinone increased the rate of disappearance of ethanol from the perfusate by about 20% and theophylline increased the rate by about 75%, the difference being statistically significant (P < 0.01). When the two drugs were added together, the level of lipolysis stimulation was not different from that with theophylline alone both at maximal and submaximal effective concentrations of drug combinations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245725 TI - Mechanisms of copper- and iron-dependent oxidative modification of human low density lipoprotein. AB - Oxidative modification of low density lipoprotein (LDL) has been suggested as a causal step in atherosclerosis, and both redox-active transition metal ions and superoxide (O2.-) have been implicated in this process. In order to determine the mechanisms of metal ion-dependent oxidation of LDL in the presence of O2.-, LDL was exposed to hypoxanthine (HX) and purified xanthine oxidase (XO) without and with added CuCl2 or Fe(3+)-citrate. Production of O2.- and hydrogen peroxide (H2O2) at pH 7.4 by the HX/XO system in the absence of metal ions was not sufficient to oxidize LDL. Preincubation of LDL with Cu2+ or Fe(3+)-citrate with subsequent removal of metal ions not tightly bound to the lipoprotein did not enable the HX/XO system to oxidize LDL. However, incubation of LDL with HX/XO and Cu2+ resulted in extensive modification of LDL. Exposure of LDL to Cu2+ alone also led to extensive modification, although the LDL was initially free of detectable amounts of lipid hydroperoxides (LOOH), i.e., < 0.005 molecules of LOOH per LDL particle. Although HX/XO and Cu2+ did not produce detectable amounts of O2.- or aqueous hydroxyl radicals (HO.), oxidation of LDL under these conditions was partially inhibited by superoxide dismutase, and completely inhibited by the HO. scavenger thiourea. In contrast to Cu(2+)-mediated oxidation of LDL, oxidation mediated by Fe(3+)-citrate was strictly dependent upon O2.-, as it was abolished by omission of the HX/XO system or by addition of superoxide dismutase to this system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245726 TI - Mutational analysis of human lipoprotein lipase by carboxy-terminal truncation. AB - We have previously reported a Trp382 (TGG)-->stop (TGA) mutation that causes familial lipoprotein lipase (LPL) deficiency. Expression study of the Trp382- >stop mutant showed that the truncated LPL was catalytically inactive with a marked reduction in the expressed mass. To investigate the minimal amino-terminal sequence of human LPL required for an active enzyme, a series of carboxy-terminal (C-terminal) truncated LPLs were expressed in vitro, and the enzyme activity, mass, and LPL mRNA levels were analyzed. The lipolytic activity showed a stepwise reduction between LPL-437 (Cys438-->stop; 68% of normal LPL activity in medium) and LPL-434 (Phe435-->stop; 3%). Without affecting LPL mRNA levels, LPL mass was reduced with the mutants not larger than LPL-437. In terms of specific activity, a significant difference was observed between LPL-436 (Lys437-->stop; 88% of that of normal LPL in medium) and LPL-435 (Val436-->stop; 22%), implying the importance of the role of Val436 in LPL action. Furthermore, our results unexpectedly showed that LPL-446 (Ser447-->stop), which is considered to be a common polymorphic form of LPL, exhibited higher activity than normal LPL (185% in medium). These results demonstrate that the C-terminal region of human LPL is closely associated with the expression of enzyme mass and catalytic activity. PMID- 8245727 TI - Abnormal activation of lipoprotein lipase by non-equilibrating apoC-II: further evidence for the presence of non-equilibrating pools of apolipoproteins C-II and C-III in plasma lipoproteins. AB - Using artificial triglyceride emulsions, we have demonstrated the presence of non equilibrating pools of apolipoproteins C-II and C-III in human plasma lipoproteins. As the concentrations of acceptor triglycerides were increased, a greater fraction of both apoC-II and apoC-III shifted away from the native plasma lipoproteins to the artificial lipid emulsions. All of the apoC-II and apoC-III in very low density and high density lipoproteins (VLDL and HDL), however, could not be removed from native plasma lipoproteins. The percent of total plasma apoC II and apoC-III that could be recovered in the VLDL and HDL density fractions varied when plasma from different individuals was used. When plasma samples from normotriglyceridemic subjects were used, HDL was the primary donor of apoCs. The percent of total plasma apoCs associated with HDL decreased from 60% to 25% for apoC-II and from 65% to 15% for apoC-III. When plasma samples from hypertriglyceridemic subjects were incubated with artificial lipid emulsions, VLDL was the primary donor of apoCs. HDL from hypertriglyceridemic subjects only accounted for 5-10% of total fasting plasma apoCs and did not contribute significantly to the final apoC contents of the artificial triglyceride emulsions. To evaluate the significance of the depletion of exchangeable apoCs from plasma HDL, we also examined the ability of control and apoC-depleted HDL to serve as activator for bovine milk lipoprotein lipase (LPL) in vitro. When HDL depleted of exchangeable apoCs were used as the source of plasma apolipoproteins for the activation of LPL in vitro, only 5-10% of the maximal activity obtained with native HDL was demonstrated. In fact, in the presence of comparable concentrations of HDL apoC-II, activation of LPL was the least with HDL which lacked exchangeable apoCs. Our data thus indicated that the presence of exchangeable apoC-II on HDL is necessary for the activation of LPL in vitro. This finding is consistent with our data that suggest that HDL from hypertriglyceridemic subjects do not stimulate LPL as well as HDL from normolipidemic subjects. PMID- 8245728 TI - Inhibitors of sterol synthesis. Chemical synthesis and properties of 3 beta hydroxy-25,26,26,26,27,27,27-heptafluoro-5 alpha-cholest-8(14)-en-15-one and 25,26,26,26,27,27,27-heptafluorocholesterol and their effects on 3-hydroxy-3 methylglutaryl coenzyme A reductase activity in cultured mammalian cells. AB - A side-chain fluorinated delta 8(14)-15-ketosterol has been synthesized from 3 beta-acetoxy-24-hydroxy-5 alpha-chol-8(14)-en-15-one (VII) as part of a program to prepare new analogs of 3 beta-hydroxy-5 alpha-cholest-8(14)-en-15-one (I), a potent regulator of cholesterol metabolism. 3 beta-Hydroxy-25,26,26,26,27,27,27 heptafluoro-5 alpha-cholest-8(14)-en-15-one (VIII) was prepared in five steps from VII in 38% overall yield. Dehydration of VII via the ortho nitrophenylselenide to the 23-ene, followed by addition of (CF3)2CFI gave 3 beta acetoxy-23R-iodo-25,26,26,26,27,27,27-heptafluoro-5 alpha-cholest-8(14)-en-15 one. Reductive deiodination with tributyltin hydride, followed by hydrolysis of the acetate gave VIII. 25,26,26,26,27,27,27-Heptafluorocholest-5-en-3 beta-ol (XXI) was prepared in eight steps in 31% overall yield from 3 alpha,6 alpha diacetoxy-5 beta-cholanic acid (XIII). Compound XIII was reduced with borane methyl sulfide to the corresponding 24-hydroxysteroid, which was converted to 3 alpha,6 alpha-diacetoxy-25,26,26,26,27,27,27-heptafluoro-5 beta-cholestane (XVIII) by reactions analogous to those developed for the preparation of VIII from VII. Conversion of XVIII via the 3 alpha,6 alpha-diol to the 3 alpha,6 alpha ditosylate, followed by heating with potassium acetate in dimethylformamide and subsequent hydrolysis gave XXI. Full 1H and 13C NMR assignments are presented for VIII, XXI, and intermediates involved in their synthesis. 13C NMR assignments for 3 alpha,6 alpha-dihydroxy-5 beta-steroids have been corrected, and stereochemical assignments were established for the side-chain methylene protons of VIII, XXI, and most synthetic intermediates. Compound VIII lowered the levels of HMG-CoA reductase activity in CHO-K1 cells and in HepG2 cells with a potency comparable to that of 3 beta-hydroxy-5 alpha-cholest-8(14)-en-15-one (I). In contrast, 25,26,26,26,27,27,27-heptafluorocholest-5-en-3 beta-ol had little or no effect on reductase activity in CHO-K1 cells. These combined results indicate that metabolism of 3 beta-hydroxy-5 alpha-cholest-8(14)-en-15-one (I) to 26- and 25 oxygenated species is not required for the suppressive action of I on the levels of HMG-CoA reductase activity in CHO-K1 cells and HepG2 cells. PMID- 8245729 TI - [13C]cholesterol as a tracer for studies of cholesterol metabolism in humans. AB - The investigation of cholesterol metabolism in humans would be facilitated by the availability of a tracer that is not radioactive. However, to be useful such a tracer must be detectable in the large pool of body cholesterol over an extended time. To meet these requirements we used synthetic [23,24,25,26,27 13C5]cholesterol with detection by isotope ratio mass spectrometry (IRMS), a technique in which the ratio of 13CO2/12CO2 can be determined with high precision in the combustion products of a biological sample. [13C]cholesterol (65 mg) and [14C]cholesterol (15 microCi) were solubilized in Intralipid and injected simultaneously (IV) into three normal subjects, and plasma samples were obtained over the ensuing 10 weeks. Cholesterol was isolated from plasma and either counted for radioactivity or combusted to CO2 and analyzed by IRMS. [14C]cholesterol and [13C]cholesterol tracer concentrations in plasma were very similar. [13C]cholesterol kinetic parameters calculated using a standard two compartment cholesterol turnover model and a new three-compartment minimal model were 103 +/- 10.5 SD % of those computed from corresponding [14C]cholesterol data. These results show that [13C5]cholesterol can be used as a practical tracer for cholesterol metabolic studies lasting many weeks. PMID- 8245730 TI - Activation waves in a model of platelet aggregation: existence of solutions and stability of travelling fronts. AB - Platelets cohere to one another to form platelet aggregates as part of the blood's clotting response. The ability of a platelet to participate in this process depends on its prior 'activation' by chemicals released into the blood plasma by other activated platelets. We study the piecewise-linear system of reaction-diffusion equations which, in one spatial dimension, describe the chemically-mediated spread of platelet activation. We establish the existence of classical solutions to this system of equations, and show that these solutions do not blow up in finite time. We also explicitly construct travelling front solutions and discuss their stability. Finally, we present numerical evidence which suggests that for a broad range of initial data with the correct limiting values at +/- infinity, the solution to the initial value problem rapidly evolves into the travelling front solution provided the front is linearly stable. PMID- 8245731 TI - Actin aggregation and embryonic epidermal wound healing. AB - Recent experiments on the response of embryonic epidermis to wounding have revealed a cable of filamentous actin at the wound edge, which may be responsible for healing (Martin and Lewis 1991, 1992). We investigate the important question of how the cable forms as a response to wounding. We modify the mechanical model of Murray and Oster (1984) to investigate the post-wounding equilibrium in the epidermal sheet. We analyse the model in both one-dimensional and radially symmetric two-dimensional geometries, to determine the parameter domain in which a solution exists. We show that in both geometries the model solutions reflect the phenomenon of the actin cable for parameter values close to one edge of this domain. We interpret these results in terms of the relative rates of intracellular reorganization of actin and myosin, and thus suggest a possible mechanism for the formation of the actin cable. PMID- 8245732 TI - Equilibrium model of a vascularized spherical carcinoma with central necrosis- some properties of the solution. AB - Properties of the solutions of a nonlinear time-independent diffusion equation are studied. The equation arises in a model of a spherically symmetric vascularized carcinoma with a central necrotic core. The boundary value problem as posed possesses a constant solution when the nutrient consumption rate and deposition rate (from the vascular network) are equal. This solution can lose uniqueness at a critical tumor dimension which corresponds to the onset of instability with respect to deviations from that uniform equilibrium state. PMID- 8245733 TI - Heterogeneous selection in subdivided populations. AB - The dynamics of allele frequencies changing under migration and heterogeneous selection in a subdivided population are investigated. Using perturbation techniques, a stationary state is obtained when migration and selection are both small. Heterogeneous selection leads to a positive correlation between values of F-statistics and heterozygosities when these are compared among sets of subdivided populations. This contrast with a negative value of the correlation obtained under Wright's classical model of homogeneous selection, and with the absence of correlation in the completely neutral situation. PMID- 8245735 TI - The ultimate transition. Death. PMID- 8245734 TI - On child abuse and the breakdown of the family. PMID- 8245736 TI - Hospice: another choice, another chance. PMID- 8245737 TI - Evoking apparitions of the deceased. PMID- 8245738 TI - Obesity and weight cycling. AB - Over 30 million Americans are overweight and in a battle to "slim down." Unfortunately for some, pounds that are lost are soon regained, leading to repeated bouts of "weight cycling." This pattern can have deleterious effects on both the physical and mental health of the individual. One must understand the causes of weight cycling to help guide the patient towards a more sensible and successful lifestyle. Emphasis on exercise, diet, and maintenance of weight loss is vital. PMID- 8245739 TI - Motor vehicle engine-related burns. PMID- 8245740 TI - The winds of change: transitions in medicine. PMID- 8245741 TI - Transitions in private practice. PMID- 8245742 TI - A physician's divorce: what's at risk? PMID- 8245743 TI - Assessment of physical education faculty students' knowledge about first aid. AB - The Physical Education Faculty students are highly exposed to different types of accidents, mainly fracture, sprains, strain, cramp, bleeding and wounds. The present study revealed that more than half of the students had correct knowledge about first aid of three injuries only out of eight namely fracture, cramp and bleeding. Lack of knowledge noted regarding cut wound, penetrating wound, falling, sprains and epistaxis. It is recommended that first aid course should be taught by specialized medical personal and the course content should include theoretical knowledge plus clinical practice of first aid in correct ways. PMID- 8245744 TI - Comparative study of staphylococcal co-agglutination and latex agglutination for detection of enterotoxigenic Escherichia coli. AB - Several biological and immunological methods have been used for detection of ETEC. In this work a comparative study was done between staph-co-agglutination and latex agglutination. From the 75 faecal samples collected from infants and children having acute diarrhea, the total number of E. Coli strains isolated were 64. Out of these 64 strains, 6 strains were found to be positive for Labile Toxin (LT) production by both latex agglutination and staphylococcal co-agglutination test. However, another 3 strains which were negative by latex agglutination were found to be positive by staph co-agglutination test. The latex agglutination test was found to be simple, easy and rapid. However this test was less sensitive in detection of ETEC. The staph-co-agglutination was also found to be simple, easy but it takes longer time (14 days). It was concluded that both tests are simple, inexpensive to perform and can be used for rapid detection of ETEC. PMID- 8245745 TI - An epidemiological study of cancer breast in greater Cairo. AB - This study was carried out to get a picture of risk factors associated with the occurrence of cancer breast. Three hundred and seventy two females (180 breast cancer patients and 192 controls) were selected from Ain Shams and Benha University Hospitals to be the subjects of this study. The results showed that the important risk factors affecting the development of cancer breast were: parity, age at first birth, age of menarche, age of the patient, family history, number of pregnancies and age of natural menopause. Mostly these factors are interrelated to each other. PMID- 8245746 TI - Effect of health education programme on knowledge about AIDS and HIV transmission in paramedical personnel working in Benha hospitals. AB - Accurate information about acquired immunodeficiency Syndrome (AIDS) and human immunodeficiency virus (HIV) is important for prevention. This study is intended to asses AIDS knowledge in paramedical personnel working in Benha Hospitals and to measure the effect of health education programme done on knowledge about AIDS in general, modes of HIV transmission and their degree of misperception about transmission through causal contact. A well designed health education programme using personal communication & visual media techniques was conducted to 238 paramedical personnel working in Benha hospitals, during the year 1992. The results of this study pointed out that the means of the scores of general AIDS knowledge and transmission knowledge were greatly and significantly improved by the end of the programme (P < 0.01). PMID- 8245747 TI - Family care of elderly problems. AB - This study was conducted to determine the elderly health problems and family care presented to them. The study was carried out in four MCH centers in Zagazig. The sample was composed of 275 women, having elderly members in their families, they were interviewed individually at the MCH center. The results showed that the main elderly physical problems were decline in daily activities, impairment of vision & digestive and urinary disorders. Psychological problems as mentioned by their families were agitation, depression and low body image. While the main identified socio-economic problems were loneliness and limited income. Regarding the family care presented to the elderly, the results showed that the low social class families had poor consideration as regards nutrition of elderly, regular physician visit, foot care, and personal hygiene. It is recommended to provide health education programs for the family about aging process, problems and needs of elderly people. As well as health education to the elderly themselves to motivate them to seek medical advice at regular intervals and integration of geriatric care in MCH centers. PMID- 8245748 TI - Cytogenetic study in idiopathic infertile males. AB - Sex chromatin investigations, including X-chromatin and Y-chromatin, was carried out on one hundred idiopathic infertile males with marked oligospermia or azoospermia. Seven cases (7%) were X-chromatin positive, 18% of the cases had aberrant Y-body (10% big Y, 7% small Y and 1% double Y). Such Y-chromosome abnormalities were frequent among azoospermic than oligospermic males. Chromosomal analysis of patients with positive X-chromatin and/or abnormalities of the Y-chromosome showed chromosomal abnormalities in the seven azoospermic cases (7%) which were X-chromatin positive. These chromosomal abnormalities are varieties of mosaic Klinefelter. Six patients (6%) were 46,XY/47,XXY mosaic and one patient (1%) was 46, XY/47,XXY/48,XXYY mosaic. In conclusion, chromosome analysis as well as sex-chromatin analysis is thus necessary in the investigation of male infertility. PMID- 8245749 TI - Risk factors of gallstone disease in a sample of patients in Benha City. AB - This study included one hundred cases of gallstone disease diagnosed by ultrasonography and a similar group of matched controls chosen from Benha University and General Hospitals. Both cases and controls were subjected to a specially designed questionnaire, clinical examination, anthropometric measurements, ultrasonography and laboratory investigations to find out some risk factors that may contribute to the development of gallstone disease. Females constituted the majority of cases (87%) while males were a minority (13%). In both sexes, the highest percentages of cases were among patients aged fifty years or more (53.8% and 33.3% for males and females respectively). Bad dietary habits including increased consumption of animal fats and frequent use of snacks between meals, obesity as indicated by body mass index and body fat percent, positive family history of gallstones and liver cirrhosis were found to be significant risk factors in the predisposition to gallstones in the present study. On the other hand, factors suggesting hormonal role in the pathogenesis of gallstones such as age of menarche, duration of menstrual life, age at first pregnancy, multiparity and duration of contraceptive pills use did not show any significant difference between female cases and controls. PMID- 8245750 TI - An epidemiologic study of the health status among elderly in Ain Shams Geriatric Unit. AB - A cross-sectional research was done aiming at the assessment of geriatric daily activities, gait and balance, besides assessment of different presentation of common diseases which occur commonly in the elderly. Special Geriatric Unit has been established in Ain Shams University Hospitals in 1984. This study included all those consulting that clinic in the period from January to April 1992. Out of 516 elderly, 270 (52.3%) were males and 246 (47.7%) were females and 478 (70%) of them were in the age group 60-70 years. Eating and wearing Clothes without help decreases with aging from 60-80 years (P < 0.001). Musculoskeletal functional disorders (joint pain and backpain) were the most frequent findings in both sexes (P < 0.001). Incontinence was the most prevalent genitourinary disorder among females Diabetes in combination with other diseases as hypertension and ischemic heart diseases was the most prevalent systemic disorder (28%). Therefore, it is recommended that these findings should be put into consideration for preventive gerontology. PMID- 8245751 TI - Incidence of nasal carriers of Staphylococcus aureus in and outside hospital environment and antibiotic sensitivity of isolated staphylococcus strains. AB - This study was carried out on 100 nasal swabs collected from medical personnel (nurses and doctors) and patients inside hospital environment and also from 50 individuals outside hospital. The swabs were inoculated on different culture media for isolation of /staphylococci which were further identified as S. aureus either by classic bacteriologic methods or by one of rapid screening test of S. aureus. The isolated strains were tested for antibiotic sensitivity to some of B Lactam antibiotics and to other antibiotics. The results showed that significantly higher percentage of coagulase + ve Staph. were isolated from newborn nursery (90%), operating theatre (71.4%) and hemodialysis unit (60%) than those isolated from intensive care unit, cancer chemotherapy, surgery, chest, internal medicine departments (25%, 26.6%, 31.2%, 33.3%, 50%) respectively. It also showed significant difference in isolation rate between persons at the hospital (patients, doctors and nurses) 44% and controls (normal population) 26%. Most isolates of coagulase + ve Staph. were resistant to penicillin G (93.2%), Streptomycin (77.3%), tetracycline (61.4%) and sensitive to cefamandole (95.4%). All coagulase+ve Staph. isolates were resistant to sulphonamide and methicillin and all sensitive to vancomycin. PMID- 8245752 TI - Collagen type III in normal and prematurely ruptured amniotic membranes. AB - This study was performed on two groups of parturient women. The first group included 20 parturient women with premature rupture of membranes (PROM) and the second group included 20 parturient women with intact membranes. From each case a specimen was taken by a sterile swab from the posterior fornix for bacteriological examination to isolate and identify both aerobic and anaerobic vaginal organisms. Also a specimen 4 x 4 cm was cut from chorioamniotic membrane at site of rupture for histological examination including routine hematoxylin and eosin staining and silver impregnation technique (Gordon & Sweet method) for identification of reticular-argyrophilic fibers (collagen III). It has been found that collagen type III was significantly decreased in amniotic membranes in cases with PROM, its decrease can be considered as a predisposing factor of PROM. The beta-hemolytic streptococci showed a significant increase in vaginal swabs taken from cases with PROM. No linking correlation was found between types of organisms in vagina and decreased collagen type III in amniotic membrane with premature rupture. Polymorphonuclear leukocytes showed no significant increase in amniotic membrane belonging to cases of PROM. PMID- 8245753 TI - The effect of praziquantel administration on the course of hepatitis B among cases with concomitant schistosome infection. AB - The present work was designed to study the course of HB virus infection among bilharzial and non-bilharzial patients and to assess the therapeutic effect of praziquantel administration on subsequent course of HB among individuals with concomitant infections. This study included 26 bilharzial cases, 14 cases with HB and 40 cases with both infections (HB and schistosomiasis). Praziquantel was administered to all bilharzial positive cases. Sera were collected from all groups prior to anti-bilharzial chemotherapy, and then later at three and six months post treatment. The results show improvement of both liver function tests and cell mediated immunity as estimated by increased mean value of CD3, CD4, helper/suppressor ratio among individuals who received praziquantel in the two groups with schistosomiasis and concomitant infection. Furthermore the individuals who lost their HB surface antigenaemia were found to have a higher mean pan T cells, CD4 values, normal helper/suppressor ratio and absence of Clc in their sera than those who retained their carrier state. The follow up of HB carriers demonstrated a higher cure rate (clearance of HBsAg) among the group with concomitant infection as compared to the group with virus hepatitis only. HBV type two infection was common among the study population accounting for 25.9% of HBsAg positive cases. 42.9% of them cleared their antigenemia after treatment with praziquantel. PMID- 8245754 TI - Root canal instruments are poorly standardized. AB - In an earlier study we demonstrated that root canal instruments vary greatly in their effectiveness in removing substrates. To further analyze the reason for this variation of results, the dimensions of the instruments used were considered of interest as their matching effectiveness may be size dependent. Nine different brands of Hedstrom files, seven brands of K files, and four brands of special files were examined. The measurements were done in a computerized measuring microscope and the data were compared with earlier reported data on machining. Although the dimensions of the instruments varied greatly, no correlation could be found between the machining and dimensions. Only one brand of files was completely within allowable size range. PMID- 8245755 TI - Biochemical and histochemical analysis of the enzyme arylsulfatase in human lesions of endodontic origin. AB - Lesions of endodontic origin are areas of inflammatory response which occur as a result of untreated disease process within the root canal system. Lysosomal hydrolytic arylsulfatase A and B have been identified as major enzymes initiating and propagating bone loss by degrading chondroitin-4-sulfate. The purpose of this investigation was to examine human lesions of endodontic origin for the presence of arylsulfatase A and B. Fifteen periapical lesions were obtained at the time of periapical surgery. The lesions were analyzed for the presence of arylsulfatases using the spectrophotometer by monitoring the liberated 4-nitrocatechol at 515-nm wavelength. The same lesions were examined histochemically using the electron microscope. Five control samples from healthy periodontal ligament were evaluated in a similar manner. The results showed higher levels of arylsulfatase A in lesions than in control tissues, and marked activity of arylsulfatase B in lesions, whereas no activity of this enzyme was detected in the control specimen. Histochemically, all lesions showed positive staining for enzyme activity, whereas the controls were negative. These findings indicate that arylsulfatase A and B play a role in the pathogenesis of human lesions of endodontic origin. PMID- 8245756 TI - Wound healing following demineralization of resected root ends in periradicular surgery. AB - The objective of this study was to determine the effect of demineralization of resected root ends on the temporal and qualitative healing of the dentoalveolar (apical attachment apparatus) and alveolar (osseous) tissues in the excisional wound site created during periradicular surgery. Root end resections to orthograde gutta-percha obturations were performed on the mandibular premolars of six mongrel dogs. Twenty-four experimental root ends were demineralized by citric acid burnishing of the resected surfaces. The remaining 24 standard treatment root ends were not demineralized. Microscopic evaluations at postsurgical intervals from 4 to 45 days revealed that the demineralized root ends were associated with more rapid and complete healing than the nondemineralized root ends. It is proposed that demineralization enhances cementogenesis, the key to dentoalveolar healing, by removing the smear layer barrier and exposing the organic component (collagen fibrils) of resected cementum and dentin. PMID- 8245757 TI - Histological evaluation of therapy using a calcium hydroxide dressing for teeth with incompletely formed apices and periapical lesions. AB - Periapical tissue response was evaluated histologically in teeth of 6-month-old dogs with incomplete root formation and periapical lesions in order to compare the effects of two calcium hydroxide-containing pastes prepared with different vehicles on induced root apex formation and repair of the periapical region. A group of teeth with no root canal dressing was used as control. After induction of the periapical lesion, the root canals were instrumented and filled with one of the two pastes (Calen and Calasept), which were renewed monthly for 90 days. At the end of the follow-up period (3 months), the animals were killed. Block sections from the actual areas were dissected free and prepared for histopathological analysis. Both calcium hydroxide pastes helped induce apical sealing and repair of the region, but the Calen paste yielded better results than Calasept. The induced mineralized tissue was predominantly of the cementoid type after the use of both pastes, but the inflammatory infiltrate was significantly less intense with the use of the Calen paste. In the control group, the absence of a root canal dressing impaired the process of periapical repair and yielded no apical root closure in any of the cases analyzed. PMID- 8245758 TI - Comparison of the sealing capabilities of Ketac-silver and extra high copper alloy amalgam when used as retrograde root canal filling. AB - Apical microleakage following reverse retrograde root filling with extra high copper amalgam alloy was compared with that following a silver-glass ionomer retrofilling. The root canals of 56 extracted, single-rooted anterior human teeth were instrumented and obturated with laterally condensed gutta-percha and zinc oxide-eugenol sealer. Each tooth was apically resected at 45 degrees to its long axis and the root surface isolated with nail varnish. Teeth were divided into three groups. The first group received extra high copper amalgam retrograde fillings, the second group was retrofilled with a silver-glass ionomer, and the control group had no retrograde root filling placed. Following immersion in 1% methylene blue dye at 37 degrees C, the roots were sectioned and dye penetration was measured using an image analyzer. The sealing effectiveness of these materials was determined by their ability to inhibit dye penetration at 24 and 48 h. The results of this study show that a silver glass-ionomer is just as effective as extra high copper amalgam in terms of sealing capability. There was no statistically significant difference between the two materials. PMID- 8245759 TI - Healing of furcation perforations in primate teeth after repair with decalcified freeze-dried bone: a longitudinal study. AB - Furcation perforations were created in 18 maxillary and mandibular molars in three adult rhesus monkeys. The perforations were located in the center of the pulp chamber floor and were 1 mm in diameter and 3 mm in depth. Teflon was used as a negative control to cover the perforation site in three teeth and the bony defects in the other 15 teeth were filled with decalcified freeze-dried bone. At the end of the 6-month experimental period the gingival tissues were clinically healthy, there was normal sulcular probing depths around all teeth, and only one tooth demonstrated furcal bone loss, radiographically. The histological picture for the Teflon and decalcified freeze-dried bone groups was very similar. There was a layer of epithelium immediately beneath the perforation site and deep to this was fibrous connective tissue and bone. Chronic inflammation was present in the connective tissue of all 3 Teflon samples but in only 2 of the 15 decalcified freeze-dried bone samples. New bone formation was not observed in any of the samples. PMID- 8245760 TI - Factors affecting apical leakage assessment. AB - This study was conducted to evaluate the influence of immediate versus delayed immersion time, and passive dye immersion versus centrifuged dye on apical leakage measurements. Eighty-four extracted human teeth with single straight canals were instrumented and divided into four experimental groups of 20 teeth each plus 2 negative and 2 positive controls. Low-temperature injection thermoplasticized gutta-percha and sealer were used to obturate the root canals. In groups A and B the filling materials were allowed to set for 72 h before the teeth were placed in India ink. In groups C and D the teeth were placed in India ink immediately after obturation. Also, in groups B and D the teeth were centrifuged in India ink for 20 min at 3,000 rpm before being immersed in ink. After 72 h in India ink, the teeth were cleared, and the linear extent of ink penetration was measured with a stereomicroscope. Statistical analysis of the data revealed no significant difference in leakage among the experimental groups whether the teeth were immersed in ink immediately after obturation or after setting of the filling materials for 72 h, and whether or not the teeth were centrifuged in ink prior to immersion. PMID- 8245761 TI - Survey of endodontic retreatment methods used to remove intraradicular posts. AB - A survey was conducted to determine the frequency and popularity of contemporary methods used in endodontic retreatment to remove intraradicular posts. Opinions and rationales were also sought for the choice of surgical management instead of nonsurgical retreatment. A questionnaire was sent to 571 active Diplomates of the American Board of Endodontics with 324 members responding (56.7%). Hand instruments, such as hemostats or special forceps, were the most commonly used, often in conjunction with ultrasonic or sonic vibration to loosen the post or break down the cement interface. A wide range of rationales was identified for the use of surgical retreatment methods, with failure to remove the post after reasonable nonsurgical efforts being the most often cited. PMID- 8245762 TI - Endodontic anesthesia in mandibular molars: a clinical study. AB - Sixty-one mandibular molar teeth with clinically manifest pulpitis, which required endodontic therapy, were studied. Twenty-seven subjects received standard inferior alveolar nerve block (IANB) with 2% lidocaine HCI with 1:100,000 epinephrine and 34 subjects received IANB with 3% mepivacaine with no vasoconstrictor. Pulpal anesthesia was assessed with dichlorodifluormethane (DDM). Subjects who gave a positive response to DDM were given a periodontal ligament injection with 2% lidocaine with 1:100,000 epinephrine. This study showed that 3% mepivacaine HCI is as effective as 2% lidocaine HCI in achieving pulpal anesthesia in mandibular molars with IANB. Of a total 61 IANB with lip anesthesia, 23 subjects required periodontal ligament injection to achieve a negative response to DDM. It was concluded that lip anesthesia is not a reliable indicator of pulpal anesthesia. The use of DDM is a reliable method of determining true pulpal anesthesia. PMID- 8245763 TI - A sonic instrument for retrograde preparation. PMID- 8245764 TI - Investigation of stimulus-secretion coupling in equine sweat gland epithelia using cell culture techniques. AB - When sweat glands isolated from samples of horse skin were explanted and cultured under favourable conditions, they could exhibit cellular outgrowth. This growth could be maintained for 2-4 weeks and these primary cultures were then disaggregated and the resultant cell suspensions used to initiate epithelial cell lines. Secretion from intact equine sweat glands is regulated by beta 2 adrenoceptors and appears to be mediated by cyclic AMP, but there is evidence that calcium may also play a role. Adrenaline could increase the cyclic AMP content of the cultured cells and this response was mediated by beta 2 adrenoceptors. Adrenaline was also able to evoke a small increase in intracellular free calcium ([Ca2+]i) but the pharmacology of this response remains obscure. Adrenaline thus activates at least two potentially important second-messenger signalling pathways which have the capacity to interact, because adrenaline-evoked cyclic AMP formation was inhibited if [Ca2+]i was raised with ionomycin. The chloride permeability of mammalian epithelial cells characteristically rises during secretion, and adrenaline could increase chloride permeability in the cultured epithelia but the cells did not contain cyclic-AMP dependent chloride channels and so this response was mediated by [Ca2+]i. PMID- 8245765 TI - Muscle temperature and swim velocity profiles during diving in a Weddell seal, Leptonychotes weddellii. AB - Locomotory muscle temperature and swim velocity profiles of an adult Weddell seal were recorded over a 21 h period. The highest temperatures occurred during a prolonged surface period (mean 37.3 degrees C, S.D. 0.16 degrees C). Muscle temperature averaged 36.8 and 36.6 degrees C (S.D. 0.25 degrees C, 0.19 degrees C) during two dive bouts and showed no consistent fluctuations between dive and interdive surface intervals. Swim velocities were also constant, near 1.3 m s-1. These data indicate that past records of low aortic temperatures (35 degrees C) during and after prolonged dives are not indicative of whole-body temperature changes, and that muscle temperature, even during dives as long as 45 min, remains near 37 degrees C. PMID- 8245766 TI - High pH in the ectoperitrophic space of the larval lepidopteran midgut. PMID- 8245767 TI - Three-year-olds' understanding of mental states: the influence of trickery. AB - This study examined the claim that children under 4 cannot conceptualize false mental states. Fifty-one children between 3;0 and 4;2 were tested in a Standard and a Trick Condition. In the Standard Condition, children were given a modified version of the Smarties task developed by Hogrefe, Wimmer, and Perner (1986). In the Trick Condition, the Smarties task was modified so that children were required to trick another person by switching the contents of a familiar box. Results revealed that a majority of subjects responded correctly to Ignorance and False Belief Questions in the Trick Condition, but not in the Standard Condition. There were no differences in children's ability to attribute ignorance and false belief. These results suggest that even young 3-year-olds have the ability to attribute mental states that differ from their own, even though this ability has proved elusive and difficult to demonstrate. PMID- 8245768 TI - Longitudinal study of the development of derived relations in an infant. AB - A normally developing child, Charlie (16 months old at the beginning and 27 months old at the end of this study), was tested several times for the derivation of relations over a period of 8 months. In a series of studies Charlie was: (1) taught to match names to pictures or pictures to names and was tested for derived relations of mutual entailment, (2) tested for retention of trained and derived relations after a 2 week delay and for the derivation of mutual entailment relations after a 1 week delay from training, (3) taught to match sounds to pictures and names to pictures and tested for mutual entailment relations and name-sound and sound-name combinatorial entailment relations, and (4) tested for the matching of a novel picture to a novel name ("nonverbal" exclusion) and for subsequent naming of the novel excluded picture ("verbal" exclusion). The results show that Charlie derived mutual entailment relations and showed nonverbal exclusion as early as 17 months. Combinatorial entailment relations and verbal exclusion emerged later. These findings lend support to the view that derivation of relations is not dependent upon sophisticated verbal abilities, and that such performances can be viewed as historically and contextually situated actions that develop over time. PMID- 8245769 TI - Processing time decreases globally at an exponential rate during childhood and adolescence. AB - Children (N = 108) from 6 to 16 years of age and adults (N = 108) were tested on four speeded tasks that included 19 different experimental conditions. The 6- to 16-year-olds' response times (RTs) increased linearly as a function of adults' RTs in those same conditions. The slope of this linear function declined exponentially with age at a rate that could be predicted accurately from values established in previous research. PMID- 8245770 TI - H-2Dd exploits a four residue peptide binding motif. AB - We have characterized the amino acid sequences of over 20 endogenous peptides bound by a soluble analog of H-2Dd, H-2Dds. Synthetic analogs corresponding to self, viral, tumor, or motif peptides were then tested for their ability to bind to H-2Dd by serologic epitope induction assays using both purified soluble protein and cell surface H-2Dd. The dominant primary sequence motif included glycine at position 2, proline at position 3, and a hydrophobic COOH terminus: leucine, isoleucine, or phenylalanine at position 9 or 10. Ancillary support for high affinity binding was contributed by a positively charged residue at position 5. Three-dimensional computer models of H-2Dds/peptide complexes, based on the crystallographic structure of the human HLA-B27/peptide complex, showed that the basic residue at position 5 was in position to form a salt bridge with aspartic acid at position 156, a polymorphic residue of the H-2Dd heavy (H) chain. Analysis of 28 such models, including 17 based on nonamer self-peptides, revealed considerable variation in the structure of the major histocompatibility complex (MHC) surrounding peptide residue 1, depending on the size and charge of the side chain. Interactions between the side chains of peptide residues 5 and 7, and 6 and 8 commonly occurred. Those peptide positions with limited sequence variability and least solvent accessibility may satisfy structural requirements for high affinity binding of the peptide to the MHC class I H chain, whereas the highly variable positions of the peptide (such as positions 4, 6, and 8) may contribute more to the T cell epitopes. PMID- 8245771 TI - Immortalized dendritic cell line fully competent in antigen presentation initiates primary T cell responses in vivo. AB - Dendritic cells (DC) can provide all the known costimulatory signals required for activation of unprimed T cells and are the most efficient and perhaps the critical antigen presenting cells in the induction of primary T cell-mediated immune responses. It is now shown that mouse cell lines with many of the features of DC can be generated using the MIB phi 2-N11 retroviral vector transducing a novel envAKR-mycMH2 fusion gene. The immortalized dendritic cell line (CB1) displays most of the morphologic, immunophenotypic, and functional attributes of DC, including constitutive expression of major histocompatibility complex (MHC) class II molecules, costimulatory molecules B7/BB1, heat stable antigen, intracellular adhesion molecule 1, and efficient antigen-presenting ability. Granulocyte/macrophage colony-stimulating factor (GM-CSF) proved to be effective in increasing MHC class II molecule expression and in enhancing presentation of native protein antigens. In comparison with macrophages, CB1 dendritic cells did not exhibit phagocytic and chemotactic activity in response to various stimuli and lipopolysaccharide activation was ineffective in inducing tumor necrosis factor alpha or interleukin 1 beta production. CB1 cells, pulsed with haptens in vitro and injected into naive mice were able to induce delayed-type hypersensitivity responses, further increased with pretreatment with GM-CSF, indicating that these cells may represent an immature, rather than a mature DC. The ability of CB1 to prime T cells in vivo could provide a tool to design novel immunization strategies. PMID- 8245772 TI - Human rheumatoid factor cross-idiotypes. IV. Studies on WA XId-positive IgM without rheumatoid factor activity provide evidence that the WA XId is not unique to rheumatoid factors and is distinct from the 17.109 and G6 XIds. AB - The WA cross-idiotype (XId) is the major XId among human monoclonal rheumatoid factors (mRF) and is almost always associated with the light (L) chain XId, 17.109, and the heavy (H) chain XId, G6. A cell line, 35G6, was cloned that bears the WA XId, but shows no reactivity with immunoglobulin G (IgG) and is negative for the 17.109 and G6 XIds. The 35G6 L chain appears to be derived from the same VKIII-JKI genes as most WA mRFs L chains. In contrast to the WA mRFs H chains in which VH1 genes are used, the 35G6 IgM expresses a VH3 gene. Sequence comparisons with other WA XId-positive mRF suggested several common structural features that may be related to the WA XId and differences that may relate to lack of IgG reactivity. Cells similar to 35G6 have previously been described in pokeweed mitogen-stimulated cell lines of peripheral blood lymphocytes from normal individuals and patients with rheumatoid arthritis and type II mixed cryoglobulinemia. These observations were confirmed, and in addition, it was shown that the majority of WA XId-positive cells in these cultures were negative for the 17.109 and G6 XIds. The presence of the WA XId in the absence of IgG reactivity suggests that the WA XId is more directly associated with an antigen specificity other than IgG, and its association with RF activity may be incidental. It is postulated that these WA XId-positive RF-negative antibodies may serve a physiologic role as natural antibodies to a pervasive pathogen, and that IgG reactivity is a consequence of somatic diversification accompanying proliferation of the WA XId-positive RF-negative cell. PMID- 8245773 TI - p120 GAP requirement in normal and malignant human hematopoiesis. AB - There is evidence to suggest that the p120 GAP (GAP), originally described as an inhibitor of p21ras, may also serve as a downstream effector of ras-regulated signal transduction. To determine whether GAP expression is required for the growth of human normal and leukemic hematopoietic cells, we used GAP antisense oligodeoxynucleotides to inhibit it and analyzed the effects of this inhibition on the colony-forming ability of nonadherent, T lymphocyte-depleted mononuclear cells and of highly purified progenitors (CD34+ MNC) obtained from the bone marrow and peripheral blood of healthy volunteers or chronic myeloid leukemia (CML, bcr-abl-positive) patients. The acute myelogenous leukemia cell line MO7, the Philadelphia BV173 cell line, and the acute promyelocytic leukemia NB4 and HL 60 cell lines were similarly examined. GAP antisense treatment inhibited colony formation from normal myelo-, erythro-, and megakaryopoietic progenitor cells as well as from CML progenitor cells. Proliferation of MO7 (growth factor-dependent) and BV173 (bcr-abl-dependent) cells, but not that of NB4 and HL-60 (growth factor independent) cells, was also inhibited, even though a specific downregulation of GAP was observed in each cell line, as analyzed by either or both mRNA and protein expression. Stimulation of MO7 cells with hematopoietic growth factors increased the expression of GAP as well as the levels of active GTP-bound p21ras. Stimulation of GAP expression was inhibited upon GAP antisense treatment. These data indicate that p120 GAP is involved in human normal and leukemic hemopoiesis and strongly suggest that GAP is not only a p21ras inhibitor (signal terminator), but also a positive signal transducer. PMID- 8245774 TI - 5-lipoxygenase and 5-lipoxygenase-activating protein are localized in the nuclear envelope of activated human leukocytes. AB - The intracellular distribution of the enzyme 5-lipoxygenase (5-LO) and 5 lipoxygenase-activating protein (FLAP) in resting and ionophore-activated human leukocytes has been determined using immuno-electronmicroscopic labeling of ultrathin frozen sections and subcellular fractionation techniques. 5-LO is a 78 kD protein that catalyzes the conversion of arachidonic acid to leukotrienes. FLAP is an 18-kD membrane bound protein that is essential for leukotriene synthesis in cells. In response to ionophore stimulation, 5-LO translocates from a soluble to a sedimentable fraction of cell homogenates. In activated leukocytes, both FLAP and 5-LO were localized in the lumen of the nuclear envelope. Neither protein could be detected in any other cell compartment or along the plasma membrane. In resting cells, the FLAP distribution was identical to that observed in activated cells. In addition, subcellular fractionation techniques showed > 83% of immunoblot-detectable FLAP protein and approximately 64% of the FLAP ligand binding activity was found in the nuclear membrane fraction. A fractionation control demonstrated that a plasma membrane marker, detected by a monoclonal antibody PMN13F6, was not detectable in the nuclear membrane fraction. In contrast to FLAP, 5-LO in resting cells could not be visualized along the nuclear envelope. Except for weak labeling of the euchromatin region of the nucleus, 5-LO could not be readily detected in any other cellular compartment. These results demonstrate that the nuclear envelope is the intracellular site at which 5-LO and FLAP act to metabolize arachidonic acid, and that ionophore activation of neutrophils and monocytes results in the translocation of 5-LO from a nonsedimentable location to the nuclear envelope. PMID- 8245775 TI - Thymus-independent positive and negative selection of T cells expressing a major histocompatibility complex class I restricted transgenic T cell receptor alpha/beta in the intestinal epithelium. AB - We demonstrate that in the mouse intestinal epithelium the selection of T lymphocytes expressing a transgenic T cell receptor alpha/beta (TCR-alpha/beta) specific for male antigen (H-Y) in the context of H-2Db depends on the differential expression of H-Y and H-2Db in situ. In H-2Db transgenic males, there is no reduction in the number of intestinal intraepithelial lymphocytes (IEL), and the four main subsets of IEL expressing TCR-alpha/beta, defined by the differential expression of CD4, CD8 alpha, and CD8 beta, are present. Moreover, the level of expression of CD8 alpha and CD8 beta on CD8+ IEL subsets is unaltered. The frequency of CD8 alpha+ IEL expressing CD8 beta, in H-2Db male mice, however, is significantly decreased and these cells do not express the transgenic TCR. In contrast, virtually all CD8 alpha+beta- IEL in the same animals express the transgenic TCR. Still, these potentially autoreactive cells are refractile to H-Y/H-2Db stimulation in vitro. Both H-2Db and H-2Dd transgenic females contain high frequencies of cells expressing the transgenic TCR among CD8 alpha+beta- and CD8 alpha+beta+ IEL. However, two possibly related phenotypic features are peculiar to H-2Db female mice. The frequency of CD8 alpha+ IEL expressing CD8 beta is increased in these mice and, while in H-2Dd females the level of the transgenic TCR alpha chain expressed on CD8 alpha+beta+ IEL is uniformly low, some of the CD8 alpha+beta+ IEL in H-2Db females express a high level of both transgenic TCR chains. It is important to note, the ability of CD8 alpha+beta+ IEL to respond to H-Y/H-2Db stimulation in vitro is restricted to those coexpressing a high level of both transgenic TCR chains. The analysis of athymic radiation chimeras using adult thymectomized recipients of distinct H-Y/H 2 haplotypes, reconstituted with bone marrow from H-2Db transgenic females, demonstrates that all IEL subsets present in unmanipulated transgenic animals develop in the absence of a thymus. These IEL are phenotypically identical to those found in unmanipulated transgenic animals sharing the H-Y/H-2 haplotype of athymic recipients. Taken together, these results demonstrate that in the absence of male antigen, expression of H-2Db in the intestinal epithelium results in the positive selection of functional IEL specific for male antigen, in situ. When both H-Y and H-2Db are expressed in the intestinal epithelium, CD8 alpha+beta+ IEL expressing the transgenic TCR are negatively selected, while the frequency of nonfunctional CD8 alpha+beta- IEL expressing the transgenic CR is increased.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8245776 TI - Folding and assembly of major histocompatibility complex class I heterodimers in the endoplasmic reticulum of intact cells precedes the binding of peptide. AB - Major histocompatibility complex (MHC) class I molecules are heterotrimers consisting of a polymorphic H chain, beta 2-microglobulin (beta 2m) and peptide. Peptides are thought to associate early during biosynthesis but the order of assembly of class I molecules from their component subunits in intact cells is not settled. We have studied the assembly of MHC class I molecules in intact cells with or without peptide transporters. MHC class I H chain/beta 2m heterodimers can be efficiently recovered only 4 min after translation and are preceded by a folding intermediate. Approximately 2 min after their formation, the class I heterodimers are loaded with peptides resulting in stable class I heterotrimers. In these in vivo studies, no evidence was obtained that peptide binding to the H chain preceded the association with beta 2m. In contrast, nonassembled class I H chains could be recovered immediately after translation, but this pool did not participate in the formation of class I molecules. PMID- 8245777 TI - V(D)J recombination in peritoneal B cells of leaky scid mice. AB - Developing lymphocytes in immune-deficient severe combined immunodeficient (scid) mice express a defective recombinase activity and rarely succeed in making an antigen receptor; those cells that do succeed account for the known B and T cell leakiness in this mutant mouse strain. To gain more insight into the nature of the scid defect, we assessed the status of heavy (H) and light (L)k, chain genes in immunoglobulin (Ig)Mk-secreting B cells from the peritoneal cavity of old leaky scid mice, the only lymphoid site where scid B cells have been routinely detected. We found these cells to be unusual in that their nonexpressed H chain alleles were either abnormally rearranged or in germline configuration (wild-type B cells generally show normal rearrangements at both H chain alleles). The VDJH junctions of the expressed alleles showed little or no nontemplated (N) addition, similar to neonatal B cells from wild-type mice. About half of the V(D)J junctions lacking N additions contained nucleotides that could have been encoded by either of the participating coding elements (VDH, DJH, or VJk), indicating that the recombination occurred between short stretches of homology. Unusually long templated (P) additions were seen in both VDJH and VJK junctions, and many recombinations appeared to involve P-based homologies. These findings suggest that: (a) B cell leakiness results from a low frequency of coding joint formation in cells expressing the defective scid recombinase activity; (b) joining of scid coding ends is facilitated when the ends contain short stretches of sequence homology, where in many cases, one of the homologous sequences results from a P addition; and (c) scid peritoneal B cells may arise early in ontogeny. PMID- 8245778 TI - Growth control or terminal differentiation: endogenous production and differential activities of vitamin A metabolites in HL-60 cells. AB - Vitamin A (retinol) is a prohormone that exerts its pleiotropic biological effects after conversion into multiple metabolites. In this report we describe the identification of three endogenous, retinolderived effector molecules, 14 hydroxy-retro-retinol (14-HRR), anhydroretinol (AR), and retinoic acid (RA) and a putative storage form of retinol, retinylesters (RE) in the human promyelocytic leukemia cell line HL-60. Exogenous application of the retinol metabolites in retinol-depleted serum-free cultures of HL-60 allowed the identification of unique cellular functions for each metabolite: 14-HRR is a growth factor for HL 60. AR is a functional antagonist of 14-HRR with growth-inhibiting activity, and RA is a potent inducer of granulocyte differentiation accompanied by growth arrest. Finally, intracellular RE serves as storage form allowing continuous production of 14-HRR when no external retinol is available. PMID- 8245779 TI - Immunoglobulin gamma 2b transgenes inhibit heavy chain gene rearrangement, but cannot promote B cell development. AB - Transgenic mice with a gamma 2b transgene were produced to investigate whether gamma 2b can replace mu in the development of B lymphocytes. Transgenic gamma 2b is present on the surface of B cells. Young transgenic mice have a dramatic decrease in B cell numbers, however, older mice have almost normal B cell numbers. Strikingly, all gamma 2b-expressing B cells in the spleen also express mu. The same is true for mice with a hybrid transgene in which the mu transmembrane and intracytoplasmic sequences replace those of gamma 2b (gamma 2b mumem). The B cell defect is not due to toxicity of gamma 2b since crosses between gamma 2b transgenic and mu transgenic mice have normal numbers of B cells. Presence of the gamma 2b transgene strongly enhances the feedback inhibition of endogenous heavy chain gene rearrangement. Light chain genes are expressed normally, and the early expression of transgenic light chains does not improve B cell maturation. When the endogenous mu locus is inactivated, B cells do not develop at all in gamma 2b transgenic mice. The data suggest that gamma 2b cannot replace mu in promoting the developmental maturation of B cells, but that it can cause feedback inhibition of heavy chain gene rearrangement. Thus, the signals for heavy chain feedback and B cell maturation appear to be different. PMID- 8245780 TI - Model for the in vivo assembly of nascent Ld class I molecules and for the expression of unfolded Ld molecules at the cell surface. AB - To characterize the process of class I assembly and maturation, we have studied the Ld molecule of the mouse. Previous studies have shown that a significant proportion of intracellular and surface Ld molecules can be detected in an alternative conformation designated Ldalt1. Nascent Ldalt molecules are non peptide ligand associated and are weakly associated with beta 2-microglobulin (beta 2m). Unexpectedly, when monoclonal antibodies were added directly to the lysis buffer, significant amounts of Ldalt/beta 2m heterodimer were detected, suggesting that beta 2m association is not necessarily sufficient to induce Ld conformation. By contrast, addition of peptide to cell lysates rapidly induced the folding of beta 2m-associated Ldalt to conformed Ld. Furthermore, the time course and dynamics of this conversion correlated precisely with peptide binding to Ld. The precursor-product relationship of Ldalt and conformed Ld was also visualized in vivo by pulse-chase analysis of BALB/c splenocytes. To investigate the factors that regulate intracellular transport of class I molecules, expression of Ld was studied in the peptide transport-deficient cell line, RMA.S Ld, and in beta 2m-/- splenocytes. In contrast to wild-type cell lines, both Ldalt and conformed Ld are poorly expressed at the cell surface of RMA.S-Ld and beta 2m-/- splenocytes. Therefore, surface expression of Ldalt is dependent upon the concomitant expression of conformed Ld molecules. To determine whether surface Ldalt molecules can result from melting of conformed Ld molecules, surface Ld molecules were loaded with several different known Ld peptide ligands. Complexes of Ld with different ligands were found to have dramatically disparate surface half-lives. Importantly, the Ld peptide complexes that turned over the most rapidly resulted in the most gain in surface Ldalt, implying that peptide dissociation can induce the accumulation of nonconformed Ld heavy chains at the cell surface. PMID- 8245781 TI - Restitution of superoxide generation in autosomal cytochrome-negative chronic granulomatous disease (A22(0) CGD)-derived B lymphocyte cell lines by transfection with p22phax cDNA. AB - The respiratory burst oxidase of phagocytes and B lymphocytes is a multicomponent enzyme that catalyzes the one-electron reduction of oxygen by NADPH. It is responsible for the O2-production that occurs when these cells are exposed to phorbol 12-myristate 13-acetate or physiologic stimuli, such as phagocytosis in phagocytes or cross-linking of surface immunoglobulin in B lymphocytes. The activity of this enzyme is greatly diminished or absent in patients with chronic granulomatous disease (CGD), an inherited disorder characterized by a severe defect in host defense against bacteria and fungi. In every CGD patient studied so far, an abnormality has been found in a gene encoding one of the four components of the respiratory burst oxidase: the membrane proteins p22phox or gp91phox which together form the cytochrome b558 protein, or the cytosolic proteins p47phox or p67phox. Autosomal recessive cytochrome-negative CGD (A22(0) CGD) is associated with mutations in the gene coding for p22phox. We report here that the capacity for O2- production and cytochrome b558 protein expression were restored to Epstein-Barr virus-transformed B lymphocytes from two A22(0) CGD patients by transfection with an expression plasmid containing a p22phox cDNA. No detectable O2- was generated by untransfected p22phox-deficient lymphocytes. The genetic reconstitution of the respiratory burst in A22(0) CGD B lymphocytes by transfer of the wild-type p22phox cDNA represents a further step towards somatic gene therapy for this subgroup of A22(0) CGD. This system will also be useful for expression of genetically engineered mutant p22phox proteins in intact cells, facilitating the structure-function analysis of cytochrome b558. PMID- 8245782 TI - Major histocompatibility complex susceptibility genes for dermatitis herpetiformis compared with those for gluten-sensitive enteropathy. AB - Dermatitis herpetiformis (DH) shares some clinical features and major histocompatibility complex (MHC) markers with gluten-sensitive enteropathy (GSE). We compared MHC haplotypes in 27 patients with DH, 35 patients with GSE, and normal controls. As in GSE, the frequencies of two extended haplotypes, [HLA-B8, SC01, DR3] and [HLA-B44, FC31, DR7], were increased in patients with DH. Distributions of fragments of extended haplotypes, consisting of some but not all of the elements of complete extended haplotypes, were analyzed to attempt to localize a susceptibility gene. Besides complete extended susceptibility haplotypes, (DR3, DQ2) and (DR7, DQ2) fragments were most common in GSE. In contrast, DH showed only a few such fragments but many instances of the fragment (SC01). The differences in distribution of these fragments in the two diseases were highly significant (P < 0.002). HLA-DQ2 and DR3 had the highest odds ratios for GSE, but the highest odds ratio for DH was for the complotype SC01. These findings suggest that the MHC susceptibility gene for DH is between class II and complotype regions, closest to the complotype, whereas that for GSE is in the class II region. PMID- 8245783 TI - The related FLT4, FLT1, and KDR receptor tyrosine kinases show distinct expression patterns in human fetal endothelial cells. AB - The growth factor receptors expressed on endothelial cells are of special interest because of their potential to program endothelial cell growth and differentiation during development and neovascularization in various pathological states, such as wound healing and angiogenesis associated with tumorigenesis. Vascular endothelial growth factor ([VEGF] also known as vascular permeability factor) is a potent mitogen and permeability factor, which has been suggested to play a role in embryonic and tumor angiogenesis. The newly cloned FLT4 receptor tyrosine kinase gene encodes a protein related to the VEGF receptors FLT1 and KDR/FLK-1. We have here studied the expression of FLT4 and the other two members of this receptor family in human fetal tissues by Northern and in situ hybridization. These results were also compared with the sites of expression of VEGF and the related placenta growth factor (PlGF). Our results reveal FLT4 mRNA expression in vascular endothelial cells in developing vessels of several organs. A comparison of FLT4, FLT1 and KDR/FLK-1 receptor mRNA signals shows overlapping, but distinct expression patterns in the tissues studied. Certain endothelia lack one or two of the three receptor mRNAs. These data suggest that the receptor tyrosine kinases encoded by the FLT gene family may have distinct functions in the regulation of the growth/differentiation of blood vessels. PMID- 8245784 TI - Identification of a dominant epitope of glutamic acid decarboxylase (GAD-65) recognized by autoantibodies in stiff-man syndrome. AB - Glutamic acid decarboxylase (GAD) is the enzyme that synthesizes the neurotransmitter gamma-aminobutyric acid (GABA) in neurons and in pancreatic beta cells. It is a major target of autoimmunity in Stiff-Man syndrome (SMS), a rare neurological disease, and in insulin-dependent diabetes mellitus. The two GAD isoforms, GAD-65 and GAD-67, are the products of two different genes. GAD-67 and GAD-65 are very similar to each other in amino acid sequence and differ substantially only at their NH2-terminal region. We have investigated the reactivity of autoantibodies of 30 Stiff-Man syndrome patients to GAD. All patient sera contained antibodies that recognize strongly GAD-65, but also GAD 67, when tested by immunoprecipitation on brain extracts and by immunoprecipitation or immunocytochemistry on cells transfected with either the GAD-65 or the GAD-67 gene. When tested by Western blotting, all patient sera selectively recognized GAD-65. Western blot analysis of deletion mutants of GAD 65 demonstrated that autoantibodies are directed predominantly against two regions of the GAD-65 molecule. All SMS sera strongly recognized a fragment contained between amino acid 475 and the COOH terminus (amino acid 585). Within this region, amino acids 475-484 and 571-585 were required for reactivity. The requirement of these two discontinuous segments implies that the epitope is influenced by conformation. This reactivity is similar to that displayed by the monoclonal antibody GAD 6, suggesting the presence of a single immunodominant epitope (SMS-E1) in this region of GAD-65. In addition, most SMS sera recognized at least one epitope (SMS-E2) in the NH2-terminal domain of GAD-65 (amino acids 1 95). The demonstration in SMS patients of a strikingly homogeneous humoral autoimmune response against GAD and the identification of dominant autoreactive target regions may help to elucidate the molecular mechanisms of GAD processing and presentation involved in GAD autoimmunity. Moreover, the reactivity reported here of GAD autoantibodies in SMS partially differs from the reactivity of GAD autoantibodies in insulin-dependent diabetes mellitus, suggesting a link between the pattern of humoral autoimmunity and the clinical condition. PMID- 8245785 TI - Dominant determinants in hen eggwhite lysozyme correspond to the cryptic determinants within its self-homologue, mouse lysozyme: implications in shaping of the T cell repertoire and autoimmunity. AB - We have studied the mouse lysozyme (ML) peptide-specific T cell repertoire in mice of five different major histocompatibility complex (MHC) class II haplotypes. 14 ML peptides were tested in a lymph node T cell proliferation assay. Upon immunization of diverse mouse strains with native ML, there was no response to any of the ML peptides tested. However, nine peptides were immunogenic, although there was no consistent pattern of reactivity toward any peptide among these strains. Thus, an autoreactive T cell repertoire directed against cryptic self(ML)-determinants exists, and it is different in mice of different MHC haplotypes. Moreover, our results demonstrate that crypticity is MHC associated and not merely a structural attribute of the determinant. On comparison of the pattern of response of various peptides of ML and that of its foreign homologue, hen eggwhite lysozyme (HEL) in H-2k, H-2b, and H-2d strains of mice, a striking correlation was evident. The stretches of amino acid sequences of determinants within HEL that were dominant in each of these three strains, almost exactly overlapped in position with those of the cryptic ML determinants against which self-reactivity was demonstrated in the same strain. These results demonstrate that the dominance-crypticity relationship between HEL and ML resulting from differential processing of these two proteins is critical in determining the response to HEL rather than the degree of sequence difference between them. These observations have important implications in the shaping of the T cell repertoire for foreign proteins and in the pathogenesis of autoimmunity. PMID- 8245786 TI - The alpha 3 domain of the Qa-2 molecule is defective for CD8 binding and cytotoxic T lymphocyte activation. AB - Qa-2 is a nonclassical class I molecule encoded by the Q7 gene within the mouse major histocompatibility complex (MHC). Results from previous experiments on Qa 2, and on a chimeric Ld molecule (LQ3) in which the alpha 3 domain is encoded by Q7b, suggested that the alpha 3 domain of Qa-2 does not carry out the functions typical of the alpha 3 domains in other classical and nonclassical class I antigens. Class I molecules that contain the Qa-2 alpha 3 domain are poorly recognized by primary cytotoxic T lymphocytes (CTLs), and do not function normally in either positive or negative selection in vivo. By employing a cell cell adhesion assay we demonstrate directly that the Qa-2 alpha 3 domain in the context of the LQ3 hybrid molecule cannot bind to human CD8, although other mouse class I alpha 3 domains bind efficiently. In addition, CD8-dependent CTL-mediated lysis of target cells, in a system which requires mouse CD8-class I alpha 3 domain interactions, is deficient in cells that express the Qa-2 alpha 3 domain. When combined with our earlier work on LQ3 transgenic mice, these results provide additional molecular support for the hypothesis that interaction with CD8 is required for both positive and negative selection of class I restricted T cells in the thymus. As the Qa-2 alpha 3 domain sequence does not differ from the previously defined minimal CD8 binding sequence of other class I molecules, these results also suggest that additional amino acids in the alpha 3 domain must be critical for CD8 binding and CTL activation. PMID- 8245787 TI - Fibrin(ogen) mediates acute inflammatory responses to biomaterials. AB - Although "biocompatible" polymeric elastomers are generally nontoxic, nonimmunogenic, and chemically inert, implants made of these materials may trigger acute and chronic inflammatory responses. Early interactions between implants and inflammatory cells are probably mediated by a layer of host proteins on the material surface. To evaluate the importance of this protein layer, we studied acute inflammatory responses of mice to samples of polyester terephthalate film (PET) that were implanted intraperitoneally for short periods. Material preincubated with albumin is "passivated," accumulating very few adherent neutrophils or macrophages, whereas uncoated or plasma-coated PET attracts large numbers of phagocytes. Neither IgG adsorption nor surface complement activation is necessary for this acute inflammation; phagocyte accumulation on uncoated implants is normal in hypogammaglobulinemic mice and in severely hypocomplementemic mice. Rather, spontaneous adsorption of fibrinogen appears to be critical: (a) PET coated with serum or hypofibrinogenemic plasma attracts as few phagocytes as does albumin-coated material; (b) in contrast, PET preincubated with serum or hypofibrinogenemic plasma containing physiologic amounts of fibrinogen elicits "normal" phagocyte recruitment; (c) most importantly, hypofibrinogenemic mice do not mount an inflammatory response to implanted PET unless the material is coated with fibrinogen or the animals are injected with fibrinogen before implantation. Thus, spontaneous adsorption of fibrinogen appears to initiate the acute inflammatory response to an implanted polymer, suggesting an interesting nexus between two major iatrogenic effects of biomaterials: clotting and inflammation. PMID- 8245788 TI - Expression and catalytic activity of the tyrosine phosphatase PTP1C is severely impaired in motheaten and viable motheaten mice. AB - Mutations in the gene encoding the phosphotyrosine phosphatase PTP1C, a cytoplasmic protein containing a COOH-terminal catalytic and two NH2-terminal Src homology 2 (SH2) domains, have been identified in motheaten (me) and viable motheaten (mev) mice and are associated with severe hemopoietic dysregulation. The me mutation is predicted to result in termination of the PTP1C polypeptide within the first SH2 domain, whereas the mev mutation creates an insertion or deletion in the phosphatase domain. No PTP1C RNA or protein could be detected in the hemopoietic tissues of me mice, nor could PTP1C phosphotyrosine phosphatase activity be isolated from cells homozygous for the me mutation. In contrast, mice homozygous for the less severe mev mutation expressed levels of full-length PTP1C protein comparable to those detected in wild type mice and the SH2 domains of mev PTP1C bound normally to phosphotyrosine-containing ligands in vitro. Nevertheless, the mev mutation induced a marked reduction in PTP1C activity. These observations provide strong evidence that the motheaten phenotypic results from loss-of-function mutations in the PTP1C gene and imply a critical role for PTP1C in the regulation of hemopoietic differentiation and immune function. PMID- 8245789 TI - Advanced glycosylation endproduct-specific receptors on human and rat T lymphocytes mediate synthesis of interferon gamma: role in tissue remodeling. AB - During normal aging and in chronic diabetes the excessive accumulation of reactive glucose-protein or glucose-lipid adducts known as advanced glycosylation endproducts (AGEs) has been shown to induce tissue dysfunction, in part through interaction with AGE-specific receptors on monocyte/macrophages and other cells. Recognizing that circulating lymphocytes trafficking through tissues interact with tissue AGEs, we searched for the expression of AGE-binding sites on peripheral blood T lymphocytes. Resting rat and human T cells bound 125I-AGE albumin with an affinity of 7.8 x 10(7) M-1, whereas, after stimulation with phytohemagglutinin (PHA) for 48 h, binding affinity increased to 5.8 x 10(8) M-1. Flow cytometric analysis of resting rat T cells using polyclonal antibodies raised against rat liver AGE-binding proteins (p60 and p90) revealed the constitutive expression of both immunoreactivities. The number of resting CD4+ and CD8+ T cells positive for anti-p60 antibody binding (34.2 and 58.5%, respectively) increased to 92 and 90% of cells after 48-h stimulation with PHA. Exposure of PHA-activated T lymphocytes to AGE-albumin enhanced expression of interferon gamma (IFN-gamma) mRNA 10-fold and induced greater elaboration of the mature protein than did exposure to unmodified protein or PHA treatment alone. These data indicate that T cells contain an inducible system of surface receptors for AGE-modified proteins, and that receptor occupancy is linked to lymphokine production. This T cell AGE-receptor system might serve to target lymphocytes to AGE-rich tissues and involve them in the regulation of tissue homeostasis either by assisting in macrophage-dependent clearance of AGE-proteins, or by exerting direct antiproliferative action on mesenchymal cells. Under conditions of excessive AGE-protein and AGE lipid accumulation (e.g., aging and diabetes), enhanced production of AGE-induced IFN-gamma may accelerate immune responses that contribute to tissue injury. PMID- 8245790 TI - Comparison of peptides bound to spleen and thymus class II. AB - In the past we and others have suggested that positive selection of developing thymocytes may depend upon interaction between the alpha beta receptors on these cells and major histocompatibility complex (MHC) proteins bound to peptides found uniquely in the selecting tissue, thymus cortical epithelium. To test this hypothesis, peptides were isolated from MHC class II proteins of spleen, thymus cortical plus medullary epithelium, or thymus cortical epithelium alone. The results showed that the major peptides bound to class II on thymus cortical epithelium were also associated with spleen class II. Some peptides could only be detected in isolates from spleen, probably because of differences in the distribution or uptake of the donor proteins between spleen and thymus. Thus, although we found some tissue-specific distribution of self-peptides, our data suggest that there are no fundamental differences among these tissues in the occupancy of class II MHC by self-peptides. These results limit hypotheses which depend on a specialized mechanism of peptide generation and/or MHC class II loading to account for the positive selection of T cells on thymic cortical epithelium. PMID- 8245791 TI - Interleukin 7-induced expression of specific T cell receptor gamma variable region genes in murine fetal liver cultures. AB - We previously reported that culture of murine fetal liver (FL) cells with interleukin 7 (IL-7) results in expression of high levels of T cell receptor (TCR) gamma transcripts by a population of cells expressing Thy-1 and Pgp-1, suggesting that IL-7 promotes the growth and/or differentiation of pre-T cells. We demonstrate herein that culture of FL cells for 7 d with IL-7 caused the rearrangement and expression of TCR gamma variable (V) region genes V gamma 4 and V gamma 6, but not V gamma 5 or V gamma 7. Since this effect was not blocked by hydroxyurea, it appeared to represent induction of expression of these genes by IL-7 rather than expansion of a preexisting positive population. We also show that IL-7 induced RAG-1 and RAG-2 mRNA expression by FL cells. These data provide evidence that specific TCR gamma/delta V region genes can be rearranged and expressed by T lineage cells before their migration to the thymus, in response to IL-7. PMID- 8245792 TI - Interleukin 10 (IL-10) inhibits the release of proinflammatory cytokines from human polymorphonuclear leukocytes. Evidence for an autocrine role of tumor necrosis factor and IL-1 beta in mediating the production of IL-8 triggered by lipopolysaccharide. AB - In this study we have examined the effects of interleukin 10 (IL-10) on polymorphonuclear leukocytes (PMN), and found that it is a potent inhibitor of tumor necrosis factor (TNF), IL-1 beta, and IL-8 secretion triggered by lipopolysaccharide (LPS). Cytokine production by phagocytosing PMN was also inhibited by IL-10, but to a lesser extent than the LPS-induced production. As shown by Northern blot analysis, IL-10 diminished the levels of TNF, IL-1 beta, and IL-8 mRNAs late after the onset of stimulation of PMN with LPS. In addition, we provide evidence that the kinetics of LPS-induced IL-8 production by PMN is composed of two distinct phases. Specifically, our experiments demonstrated that in the first phase, the production of IL-8 is a process directly induced by LPS that lasts for some hours. After this early wave, a second phase begins that is sustained and leads to an elevated production of IL-8 that appears to be due to the endogenous release of TNF and IL-1 beta. This second wave can in fact be blocked by anti-TNF and anti-IL-1 beta neutralizing antibodies, and by IL-10 as the consequence of its downregulatory effects on TNF and IL-1 beta release. Taken together, these findings identify novel biological actions of IL-10 as a suppressor of the inflammatory response. PMID- 8245793 TI - The synaptic vesicle-associated protein amphiphysin is the 128-kD autoantigen of Stiff-Man syndrome with breast cancer. AB - Stiff-Man syndrome (SMS) is a rare disease of the central nervous system (CNS) characterized by progressive rigidity of the body musculature with superimposed painful spasms. An autoimmune origin of the disease has been proposed. In a caseload of more than 100 SMS patients, 60% were found positive for autoantibodies directed against the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD). Few patients, all women affected by breast cancer, were negative for GAD autoantibodies but positive for autoantibodies directed against a 128-kD synaptic protein. We report here that this antigen is amphiphysin. GAD and amphiphysin are nonintrinsic membrane proteins that are concentrated in nerve terminals, where a pool of both proteins is associated with the cytoplasmic surface of synaptic vesicles. GAD and amphiphysin are the only two known targets of CNS autoimmunity with this distribution. This finding suggests a possible link between autoimmunity directed against cytoplasmic proteins associated with synaptic vesicles and SMS. PMID- 8245794 TI - Double-negative T cells from MRL-lpr/lpr mice mediate cytolytic activity when triggered through adhesion molecules and constitutively express perforin gene. AB - The lpr gene induces in mice, accumulation of large numbers of CD4-CD8- (double negative [DN]) T lymphocytes which bear adhesion molecules not characteristic of normal resting T cells. These cells fail to acquire interleukin 2 (IL-2) receptors, produce IL-2, and proliferate when activated with mitogens or monoclonal antibodies (mAbs) against the T cell receptor (TCR). Because of these poor functions in vitro, the nature and significance of DN T cells in the autoimmune disease process is not clear. In the current study, we describe a surprising finding that mAbs against CD3-TCR-alpha/beta complex can strongly trigger the lytic activity of the DN T cells to induce redirected lysis of Fc receptor-positive targets. Similar redirected lysis was also inducible using mAbs against CD44 and gp90MEL-14, molecules involved in the binding of lymphocytes to endothelial cells. The spontaneous cytotoxic potential of the DN T cells was further corroborated by demonstrating that the lpr DN T cells constitutively transcribed perforin gene but failed to express granzyme A. The current study suggests that DN T cells are capable of mediating lysis of autologous cells bearing the specific ligands for adhesion molecules involved in the signaling of cytotoxicity. These findings provide a novel insight into the functional significance of DN T cells in lpr mice and their potential role in the pathogenesis of autoimmune disease. PMID- 8245795 TI - Disseminated tuberculosis in interferon gamma gene-disrupted mice. AB - The expression of protective immunity to Mycobacterium tuberculosis in mice is mediated by T lymphocytes that secrete cytokines. These molecules then mediate a variety of roles, including the activation of parasitized host macrophages, and the recruitment of other mononuclear phagocytes to the site of the infection in order to initiate granuloma formation. Among these cytokines, interferon gamma (IFN-gamma) is believed to play a key role is these events. In confirmation of this hypothesis, we show in this study that mice in which the IFN-gamma gene has been disrupted were unable to contain or control a normally sublethal dose of M. tuberculosis, delivered either intravenously or aerogenically. In such mice, a progressive and widespread tissue destruction and necrosis, associated with very high numbers of acid-fast bacilli, was observed. In contrast, despite the lack of protective immunity, some DTH-like reactivity could still be elicited. These data, therefore, indicate that although IFN-gamma may not be needed for DTH expression, it plays a pivotal and essential role in protective cellular immunity to tuberculosis infection. PMID- 8245796 TI - Induction and function of vascular adhesion protein-1 at sites of inflammation. AB - Emigration of leukocytes from the blood into the tissues is critical in controlling lymphocyte patrolling in different lymphatic organs and in leukocyte accumulation at sites of inflammation. During the first stage of the extravasation process, leukocytes bind to the endothelial lining of vessels. At the molecular level, several adhesion molecules on leukocytes and endothelial cells function as receptor-ligand pairs in mediating this dynamic interaction. Recently, we have identified a novel human endothelial cell molecule, vascular adhesion protein 1 (VAP-1), that mediates lymphocyte binding (Salmi, M., and S. Jalkanen. 1992. Science [Wash. DC] 257:1407). VAP-1 was initially characterized by mAb 1B2 which inhibits lymphocyte adhesion to high endothelial venules (HEV) and to purified VAP-1 protein. Here we report the location and function of VAP-1 in normal and inflamed tissues in humans. VAP-1 is abundant in HEV of lymphatic organs belonging to the peripheral lymph node system, but considerably less is expressed in vessels of mucosa-associated lymphatic tissues. A subset of venules in most normal nonlymphatic tissues like skin, brain, kidney, liver, and heart is also VAP-1 positive. In addition to vessels, VAP-1 is distributed on a few other cell types, most notably in dendritic-like cells of germinal centers. At sites of inflammation, such as in inflammatory bowel diseases and chronic dermatoses, expression of VAP-1 is clearly increased. The induced VAP-1 is functional, since mAb 1B2 inhibits lymphocyte binding to inflamed lamina propria venules by approximately 60%. Thus VAP-1 is an endothelial adhesion molecule that under normal conditions is expressed mainly in HEV of lymphatic tissues. However, expression of functional VAP-1 in vivo is upregulated during an inflammatory reaction at other sites as well. Inducibility of VAP-1 suggests that it may play a significant role, not only in recirculation of lymphocytes, but also in controlling entry of leukocytes into sites of inflammation. PMID- 8245797 TI - Effect of dietary supplementation with n-9 eicosatrienoic acid on leukotriene B4 synthesis in rats: a novel approach to inhibition of eicosanoid synthesis. AB - Studies were undertaken to assess the biochemical effects of dietary supplementation with n-9 eicosatrienoic acid (ETrA), an arachidonic acid analogue that is normally present in cell membranes at very low levels but is raised in the presence of essential fatty acid deficiency (EFAD). The incorporation of dietary ETrA into rat neutrophils and its effect on A23187-stimulated 5 lipoxygenase metabolism in these cells was examined; in addition, the effect of ETrA was compared with that of another arachidonic acid analogue, eicosapentaenoic acid (EPA), which is known to accumulate in cell membranes and inhibit synthesis of leukotriene B4 (LTB4) a product of the 5-lipoxygenase metabolic pathway. Rats were fed a defined diet that was sufficient in essential fatty acids and that contained EPA or ETrA (0.014% of energy) or no added fatty acid, for 3 wk. In the cells from ETrA-fed rats, LTB4 synthesis was inhibited relative to control values, but synthesis of the other products of 5-lipoxygenase metabolism, 5-hydroxyeicosatetraenoic acid (5-HETE) and the all-trans isomers of LTB4, were not inhibited. This pattern indicates inhibition of LTA hydrolase in ETrA-fed rats. In EPA-fed rats, there was inhibition of LTB4 and the all-trans isomers of LTB4, but there was no inhibition of 5-HETE. This pattern indicates inhibition of LTA synthase in EPA-fed rats. The results establish that dietary ETrA effectively inhibits synthesis of the inflammatory mediator, LTB4, and suggest that ETrA may confer antiinflammatory benefits similar to those observed with EFAD or dietary fish oil (which contains EPA). Because ETrA is substantially less unsaturated than EPA, it can be expected to have greater chemical stability, which could be an important practical advantage when used as a dietary constituent or supplement. PMID- 8245798 TI - Drug samples. PMID- 8245799 TI - Physician satisfaction. PMID- 8245800 TI - Assisted suicide. PMID- 8245801 TI - RBRVS. PMID- 8245802 TI - RBRVS. PMID- 8245803 TI - Concurrent care. PMID- 8245804 TI - The emperor's new clothes. PMID- 8245805 TI - Do antidepressant medications relieve chronic low back pain? AB - BACKGROUND: Antidepressant medications are commonly prescribed for patients with chronic low back pain. A literature synthesis was performed to determine whether antidepressants are more effective than placebos in decreasing pain, disability, depression, and analgesic medication use in such patients. METHODS: English language journal articles were identified from MEDLINE and PsycLIT databases, bibliographies, and inquiries to researchers and drug companies. Articles were included if they reported data from placebo-controlled or drug comparison trials of antidepressants for patients with low back pain. Six articles met these criteria. RESULTS: Three studies compared the effects of antidepressants and placebos on pain; two found no difference and one found a trend toward superiority of imipramine for patient-rated symptoms but no difference in investigator ratings. Effects on functional disability were examined in three antidepressant-placebo comparisons; only one found the antidepressant to be more effective. Antidepressant effects of an antidepressant vs placebo were compared in three studies; none found a significant difference. Effects on analgesic medication use were compared in three studies; one found amitriptyline to be superior and the others found no difference. Serious methodologic flaws characterized all six studies, and insufficient reporting of data precluded meta analysis. CONCLUSIONS: The literature has not demonstrated that antidepressants are superior to placebos in improving low back pain or related problems. However, further randomized controlled trials are needed to determine whether antidepressants are useful for low back pain. PMID- 8245806 TI - Does family practice at residency teaching sites reflect community practice? AB - BACKGROUND: Family medicine has aspired to train residents and conduct research in settings that closely resemble community practice. The purpose of this study was to compare the patient characteristics of the ambulatory teaching centers of a consortium of seven community-based university-affiliated family practice residency programs in northeast Ohio with the National Ambulatory Medical Care Survey (NAMCS) results for family physicians (FPs) and general practitioners (GPs). METHODS: Ninety-eight faculty and resident physicians at the residency training site of the Northeastern Ohio Universities College of Medicine collected data on all ambulatory patient visits (N = 1498) for one randomly chosen week between July 1, 1991, and June 30, 1992. We compared these data with patient visits reported in the 1990 NAMCS for FPs and GPs. RESULTS: The residency training sites saw slightly more children, women, blacks, and Medicare and Medicaid patients. The most common reason for an office visit in both populations was an undifferentiated symptom. Fifteen of the top 20 "reason for visit" codes were identical, as were 14 of the top 20 diagnoses. More preventive and therapeutic services were offered or performed at our residency training sites but fewer diagnostic services were performed. There were fewer consultations requested at our residency training sites but similar hospitalization rates for patients. The mean duration of visit differed by only 1 minute. CONCLUSIONS: The residency training sites of the Northeastern Ohio Universities College of Medicine provide patient care opportunities similar to those found in a national survey of family and general practitioners. PMID- 8245807 TI - Patient attitudes toward rooming with persons with HIV infection. AB - BACKGROUND: Early in the HIV epidemic, hospitals developed strict isolation policies for patients with HIV infection, some of which have not been revised. The objectives of this study were to examine patient attitudes about rooming with persons with various medical conditions, including HIV, and to assess their knowledge about the transmission of HIV. METHODS: One hundred four inpatients at a university hospital were surveyed by means of a structured interviewer administered questionnaire. Patients were asked about preference for a single or double room, and about their objections to rooming with patients with HIV infection and other medical conditions. The questionnaire also examined subject's knowledge about the transmission of HIV. RESULTS: Of 104 inpatients surveyed, 55% objected to rooming with an HIV-seropositive patient. Patients who objected to rooming with an HIV-seropositive patient were also more likely to object to rooming with a disfigured patient (relative risk = 1.5; 95% CI, 1.1 to 2.2), or with a demented patient (relative risk = 1.7; 95% CI, 1.0 to 2.9). Also, patients who objected to rooming with an HIV-seropositive patient had greater misconceptions about the transmissibility of HIV infection. CONCLUSIONS: A significant proportion of patients reported an unwillingness to room with patients with HIV infection, but also had misconceptions about the transmissibility of HIV. Current rooming policies may perpetuate misconceptions about the possibility of causal transmission of HIV. PMID- 8245808 TI - Office systems for promoting screening mammography. A survey of primary care practices. AB - BACKGROUND: Office tracking, scheduling, and reminder systems have been shown to improve utilization of screening mammography, but little is known about the use of these systems by primary care physicians. METHODS: We surveyed 132 primary care and obstetrics and gynecology practices affiliated with an independent practice association model health maintenance organization in central Massachusetts to determine their use of reminder, scheduling, and follow-up systems, and education and counseling services aimed at increasing screening mammography rates. RESULTS: The use of chart flags to remind physicians of a patient's need for mammography screening was reported by 30% of practices. Thirty one percent reported the use of flow sheets, and 27% reported the use of mail or telephone patient reminders. At least one of these three systems was used by 57% of the practices, whereas 43% reported having none of these three systems. Variations in the use of these office systems were related to specialty type, physician number, and clinical staffing. The majority of practices (77%) reported using written educational materials, and 42% offered prevention counseling with nonphysician staff. Very few offices (8%) reported using mail or telephone reminders for previously scheduled appointments. CONCLUSIONS: Despite the proven effectiveness of reminder systems for screening mammography, many practices do not have a system in place. Promotion of reminder systems in primary care practices could have a substantial impact on mammography utilization. PMID- 8245809 TI - Adequacy of prenatal care among inner-city women. AB - BACKGROUND: Lack of prenatal care is a well-recognized risk factor for infant mortality and low birthweight. This study was conducted to identify factors that facilitate or inhibit access to prenatal care among low-income inner-city women. METHODS: A case-control interview study was conducted with women during their postpartum hospitalization at a midwestern inner-city hospital. Fifty-eight women who had received no prenatal care and 71 women who had received markedly inadequate prenatal care were compared with 123 controls who had received intermediate or adequate prenatal care. RESULTS: The majority of subjects were minorities, single, had low incomes, and were in the Medicaid program. Subjects' median age was 23 years and median parity 2, and the majority had not completed high school. Inadequate prenatal care was independently associated with the following variables (adjusted odds ratios): lack of any insurance, including Medicaid (5.3), being a smoker (3.8), being homeless (2.7), being black (2.5), not being worried what the physician or nurse might say (2.4), not using contraception (2.1), having a household income of less than $400 a month (1.8), being ashamed or afraid of the pregnancy or the physician (1.4), having transportation problems (1.3), and level of education (0.8). CONCLUSIONS: Practical factors related to poverty are substantial barriers to obtaining prenatal care. Comprehensive approaches to prenatal services that address these barriers may be more effective in facilitating adequate prenatal care among low income women. PMID- 8245810 TI - Follow-up of abnormal Papanicolaou smears among women of different races. AB - BACKGROUND: Current information about racial differences in the rate of cervical abnormalities is incomplete, and there are few data about racial differences in compliance with follow-up and treatment. The purpose of this study was to investigate the frequency and follow-up of abnormal Pap smear findings in white, black, and Southeast Asian women. METHODS: The charts of women who attended a St Paul family practice residency clinic and who had abnormal Papanicolaou (Pap) smear results between January 1, 1989, and September 1, 1992, were reviewed, and information about age, race, insurance, Pap smear findings, diagnostic studies, and treatment procedures was recorded. RESULTS: Of 1794 women who had Pap smears during this period, 190 (10.6%) had abnormal results, with a diagnosis of atypia, dysplasia, or carcinoma. The rate of abnormality was greater for black women (16.4%) than for Southeast Asian (6.1%) and white women (11.6%); however, the proportion of abnormal Pap smears that showed moderately severe or worse changes was greater for Southeast Asians than for whites (30.6% vs 14.3%, P < .05). Southeast Asian women with abnormal Pap smears were also less likely than whites and blacks to follow through with recommended diagnostic and treatment procedures. CONCLUSIONS: Southeast Asian women in this study were less likely than white and black women to comply with recommended follow-up diagnostic and treatment procedures for cervical disease. PMID- 8245811 TI - Trauma in pregnancy. AB - Trauma is the leading cause of nonobstetric maternal morbidity and mortality in this country. Maternal survival does not guarantee fetal survival, even in cases of apparently minor trauma. The injured pregnant patient presents unique diagnostic and therapeutic challenges. Physicians who make obstetrics or emergency medicine part of their practice must be aware of these unique problems. Prevention of traumatic injury should remain the focus of office practice. PMID- 8245812 TI - Advances in Alzheimer's disease. A review for the family physician. AB - Alzheimer's disease accounts for approximately two thirds of all cases of dementia in the United States and $90 billion in health care costs annually. Clinical and laboratory diagnostic tools have been refined so that clinicians now can diagnose Alzheimer's disease with up to 90% accuracy. Criteria for clinical diagnosis have been outlined by a work group of the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association. Key diagnostic tools include a complete patient history, mental status testing, and a thorough diagnostic workup to exclude the possibility of a reversible disease mimicking the symptoms of Alzheimer's disease. Currently, management of Alzheimer's disease involves a two pronged approach: behavioral-supportive care and pharmacologic control of disruptive behavioral symptoms. In the future, drug therapy may be available to maintain memory and cognitive function. Cholinesterase inhibitors, which block the decrease in choline acetyltransferase activity associated with Alzheimer's disease, appear promising. The realistic goal of health care providers at the present time, however, should be symptom control rather than disease reversal. PMID- 8245813 TI - Multiple pneumonias in a man infected with HIV. AB - There are many pathogens responsible for pneumonia in persons infected with HIV. This case report describes a patient with pneumonias diagnosed sequentially and caused by Pneumocystis carinii, Mycobacterium gordonae, and Coccidioides immitis. It demonstrates the importance of pursuing a definitive or additional diagnosis in HIV-related pulmonary disease when the response to empiric therapy or to treatment of an identified pathogen is suboptimal. PMID- 8245814 TI - Determination of gestational opiate, nicotine, benzodiazepine, cocaine and amphetamine exposure by hair analysis. AB - Hair samples were collected at time of delivery from 57 neonates whose mothers were known users of heroin (9 cases), nicotine (34 cases), benzodiazepines (11 cases), cocaine (2 cases) and amphetamine (1 case). In all cases, the corresponding drug was found in neonatal hair from the infants, with concentrations in the range 0.61-3.47 ng/mg (morphine), 0.15-11.80 ng/mg (nicotine), 3.36-17.55 ng/mg (diazepam), 0.78-31.83 ng/mg (oxazepam), 0.71-2.47 ng/mg (benzoylecgonine) and 1.21 ng/mg (amphetamine). Results are considered in the light of the existing literature and comparison with adults. PMID- 8245815 TI - ABO, phosphoglucomutase and erythrocyte acid phosphatase typing of blood samples containing added fluoride. AB - Blood samples containing approximately 2% and 4% sodium fluoride have been stored at 4 degrees C for up to 5 months and 4 weeks respectively. ABO typing by absorption-elution was successful on stains made from almost all of the samples. PGM subtyping by isoelectric focusing was successful on most samples, with clearer results from whole blood than from stains. Failure to type appears to be slightly more common at the higher preservative level and yields no result rather than an erroneous one. Apparent phenotypic changes in the 1+ and 1- bands have been seen after 5 months storage. EAP typing by isoelectric focusing was successful on most samples, again with clearer results from whole bloods than from stains. Possible errors can occur in samples containing C bands, but it is not clear whether they are caused by the presence of preservative. The results obtained are illustrated by two casework examples. PMID- 8245816 TI - A simplified method for typing haemoglobin using ultrathin-layer isoelectric focusing. AB - Haemoglobin alleles were identified by isoelectric focusing using ultrathin-layer polyacrylamide gels, containing pH 3.0-10.0 ampholytes. The run parameters were chosen to co-ordinate with other isoelectric focusing methods currently used by the authors' laboratory. The method devised separated the A, F, S, and C alleles. PMID- 8245817 TI - Perchlorate enhances transmission in skeletal muscle excitation-contraction coupling. AB - The effects of the anion perchlorate (present extracellularly at 8 mM) were studied on functional skeletal muscle fibers from Rana pipiens, voltage-clamped in a Vaseline gap chamber. Established methods were used to monitor intramembranous charge movement and flux of Ca release from the sarcoplasmic reticulum (SR) during pulse depolarization. Saponin permeabilization of the end portions of the fiber segment (Irving, M., J. Maylie, N. L. Sizto, and W. K. Chandler. 1987. Journal of General Physiology. 89:1-41) substantially reduced the amount of charge moving during conventional control pulses, thus minimizing a technical error that plagued our previous studies. Perchlorate prolonged the ON time course of charge movement, especially at low and intermediate voltages. The OFFs were also made slower, the time constant increasing twofold. The hump kinetic component was exaggerated by ClO4- or was made to appear in fibers that did not have it in reference conditions. ClO4- had essentially no kinetic ON effects at high voltages (> or = 10 mV). ClO4- changed the voltage distribution of mobile charge. In single Boltzmann fits, the midpoint potential V was shifted 20 mV and the steepness parameter K was reduced by 4.7 mV (or 1.78-fold), but the maximum charge was unchanged (n = 9). Total Ca content in the SR, estimated using the method of Schneider et al. (Schneider, M. F., B. J. Simon, and G. Szucs. 1987. Journal of Physiology. 392:167-192) for correcting for depletion, stayed constant over tens of minutes in reference conditions but decayed in ClO4- at an average rate of 0.3 mumol/liter myoplasmic water per s. ClO4- changed the kinetics of release flux, reducing the fractional inactivation of release after the peak. ClO4- shifted the voltage dependence of Ca release flux. In particular, the threshold voltage for Ca release was shifted by about -20 mV, and the activation of the steady component of release flux was shifted by > 20 mV in the negative direction. The shift of release activation was greater than that of mobile charge. Thus the threshold charge, defined as the minimum charge moved for eliciting a detectable Ca transient, was reduced from 6 nC/microF (0.55, n = 7) to 3.4 (0.53). The average of the paired differences was 2.8 (0.33, P < 0.01). The effects of ClO4- were then studied in fibers in modified functional situations. Depletion of Ca in the SR, achieved by high frequency pulsing in the presence of intracellular BAPTA and EGTA, simplified but did not eliminate the effects of ClO4-.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8245818 TI - Effects of perchlorate on the molecules of excitation-contraction coupling of skeletal and cardiac muscle. AB - To understand the nature of the transmission process of excitation-contraction (EC) coupling, the effects of the anion perchlorate were investigated on the voltage sensor (dihydropyridine receptor, DHPR) and the Ca release channel (ryanodine receptor, RyR) of the sarcoplasmic reticulum (SR). The molecules, from rabbit skeletal muscle, were either separated in membrane vesicular fractions or biochemically purified so that the normal EC coupling interaction was prevented. Additionally, the effect of ClO4- was investigated on L-type Ca2+ channel gating currents of guinea pig ventricular myocytes, as a native DHPR not in the physiological interaction of skeletal muscle. At 20 mM, ClO4- had minor effects on the activation of ionic currents through Ca channels from skeletal muscle transverse tubular (T) membranes fused with planar bilayers: a +7-mV shift in the midpoint voltage, V, with no change in kinetics of activation or deactivation. This is in contrast with the larger, negative shift that ClO4- causes on the distribution of intramembrane charge movement of skeletal muscle. At up to 100 mM it did not affect the binding of the DHP [3H]PN200-110 to triad-enriched membrane fractions (TR). At 8 mM it did not affect the kinetics or the voltage distribution of gating currents of Ca channels in heart myocytes. These negative results were in contrast to the effects of ClO4- on the release channel. At 20 mM it increased several-fold the open probability of channels from purified RyR incorporated in planar bilayers and conducting Ba2+, an effect seen on channels first closed by chelation of Ca2+ or by the presence of Mg2+. It significantly increased the initial rate of efflux of 45Ca2+ from TR vesicles (by a factor of 1.75 at 20 mM and 4.5 at 100 mM). ClO4- also increased the binding of [3H]ryanodine to TR fractions. The relative increase in binding was 50-fold at the lowest [Ca2+] used (1 microM) and then decayed to much lower values as [Ca2+] was increased. The increase was due entirely to an increase in the association rate constant of ryanodine binding. The chaotropic ions SCN- and I- increased the association rate constant to a similar extent. The binding of ryanodine to purified RyR protein reconstituted into liposomes had a greater affinity than to TR fractions but was similarly enhanced by ClO4-. The reducing agent dithiothreitol (5 mM) did not reduce the effect of ClO4-, and 5% polyethylene glycol, with an osmolarity equivalent to 20 mM ClO4-, did not change ryanodine binding.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8245819 TI - An allosteric model of the molecular interactions of excitation-contraction coupling in skeletal muscle. AB - A contact interaction is proposed to exist between the voltage sensor of the transverse tubular membrane of skeletal muscle and the calcium release channel of the sarcoplasmic reticulum. This interaction is given a quantitative formulation inspired in the Monod, Wyman, and Changeux model of allosteric transitions in hemoglobin (Monod, J., J. Wyman, and J.-P. Changeux. 1965. Journal of Molecular Biology. 12:88-118), and analogous to one proposed by Marks and Jones for voltage dependent Ca channels (Marks, T. N., and S. W. Jones. 1992. Journal of General Physiology. 99:367-390). The allosteric protein is the calcium release channel, a homotetramer, with two accessible states, closed and open. The kinetics and equilibrium of this transition are modulated by voltage sensors (dihydropyridine receptors) pictured as four units per release channel, each undergoing independent voltage-driven transitions between two states (resting and activating). For each voltage sensor that moves to the activating state, the tendency of the channel to open increases by an equal (large) factor. The equilibrium and kinetic equations of the model are solved and shown to reproduce well a number of experimentally measured relationships including: charge movement (Q) vs. voltage, open probability of the release channel (Po) vs. voltage, the transfer function relationship Po vs. Q, and the kinetics of charge movement, release activation, and deactivation. The main consequence of the assumption of allosteric coupling is that primary effects on the release channel are transmitted backward to the voltage sensor and give secondary effects. Thus, the model reproduces well the effects of perchlorate, described in the two previous articles, under the assumption that the primary effect is to increase the intrinsic tendency of the release channel to open, with no direct effects on the voltage sensor. This modification of the open-closed equilibrium of the release channel causes a shift in the equilibrium dependency of charge movement with voltage. The paradoxical slowing of charge movement by perchlorate also results from reciprocal effects of the channel on the allosterically coupled voltage sensors. The observations of the previous articles plus the simulations in this article constitute functional evidence of allosteric transmission. PMID- 8245820 TI - Visual pigment bleaching in isolated salamander retinal cones. Microspectrophotometry and light adaptation. AB - Visual pigment bleaching desensitizes rod photoreceptors greatly in excess of that due to loss of quantum catch. Whether this phenomenon also occurs in cone photoreceptors was investigated for isolated salamander red-sensitive cones. In parallel experiments, (a) visual pigment depletion by steps of bleaching light was measured by microspectrophotometry, and (b) flash sensitivity was measured by recording light-sensitive membrane current. In isolated cones, visual pigment bleaching permanently reduced flash sensitivity significantly below that due to the reduction in quantum catch, and there was little spontaneous recovery of visual pigment. The "extra" desensitization due to bleaching was most prominent up to bleaches of approximately 80% visual pigment and reached a level approximately 1 log unit beyond that due to loss of quantum catch. At higher bleaches, the effect of loss of quantum catch became more important. Bleaching did not greatly reduce the maximum light-suppressible membrane current. A 99% reduction of the visual pigment permanently reduced the circulating current by only 30%. Visual pigment bleaching speeded up the kinetics of dim flash responses. All electrical effects of bleaching were reversed on exposure to 11 cis retinal, which probably caused visual pigment regeneration. Light adaptation in photopic vision is known to involve significant visual pigment depletion. The present results indicate that cones operate with a maintained circulating current even after a large pigment depletion. It is shown how Weber/Fechner behavior may still be observed in photopic vision when the contributions of bleaching to adaptation are included. PMID- 8245821 TI - Ascorbic acid modulation of calcium channels in pancreatic beta cells. AB - We have studied the effect of ascorbic acid on voltage-dependent calcium channels in pancreatic beta cells. Using the whole-cell and perforated-patch variants of the patch clamp technique to record calcium tail currents, we have shown that the slowly deactivating (SD) calcium channel, which is similar to the T-type channel in other cells, is inhibited in a voltage-dependent manner by ascorbic acid (AA). The other channels that carry inward current in beta cells, FD calcium channels and sodium channels, are unaffected by AA. Ascorbic acid causes a voltage dependent decrease in the magnitude of the SD channel conductance which can be explained by the hypothesis that approximately 50-60% of the channels have their voltage dependence shifted by approximately 62 mV in the depolarizing direction. Thus, ascorbate appears to modify only a fraction of the SD channels. The activation kinetics of the ascorbate-modified channels are slower than control channels in a manner that is consistent with this hypothesis. Deactivation and inactivation kinetics are unaffected by ascorbate. These effects of ascorbate require metal ions, and it appears that some of the activity of ascorbate is due to a product of its metal catalyzed oxidation, perhaps dehydroascorbate. PMID- 8245822 TI - Regulation of Ca2+ current in frog ventricular cardiomyocytes by guanosine 5' triphosphate analogues and isoproterenol. AB - Calcium currents (ICa) were measured in frog ventricular myocytes using the whole cell patch clamp technique and a perfused pipette. To gain insight into the role of G proteins in the regulation of ICa in intact cells, the effect of internal perfusion with hydrolysis-resistant GTP analogues, guanylyl 5'-imidodiphosphate (GppNHp) or guanosine 5'-thiotriphosphate (GTP gamma S), on ICa stimulated by isoproterenol (Iso) or forskolin (Forsk) was examined. Significant differences were observed between the effects of the two GTP analogues. Internal perfusion of GppNHp resulted in a near-complete (approximately 80%) and irreversible inhibition of Iso-stimulated ICa. In contrast, internal perfusion with GTP gamma S resulted in only a partial (approximately 40%) inhibition of Iso- or Forsk stimulated ICa. The fraction of the current not inhibited by GTP gamma S remained persistently elevated after the washout of Iso but declined to basal levels upon washout of Forsk. Excess internal GTP or GppNHp did not reduce the persistent ICa. Internal adenosine 5'-thiotriphosphate (ATP gamma S) mimicked the GTP gamma S-induced, persistent ICa. GppNHp sometimes induced a persistent ICa, but only if GppNHp was present at high concentration before Iso exposure. Inhibitors of protein kinase A inhibited both the GTP gamma S- and ATP gamma S-induced, persistent ICa. We conclude that: (a) GTP gamma S is less effective than GppNHp in inhibiting adenylyl cyclase (AC) via the inhibitory G protein, Gi; and (b) the persistent ICa results from a long-lived Gs-GTP gamma S complex that can activate AC in the absence of Iso. These results suggest that different hydrolysis resistant nucleotide analogues may behave differently in activating G proteins and imply that the efficacy of G protein-effector molecule interactions can depend on the GTP analogue with which the G protein is activated. PMID- 8245823 TI - The role of ATP in swelling-stimulated K-Cl cotransport in human red cell ghosts. Phosphorylation-dephosphorylation events are not in the signal transduction pathway. AB - Volume-sensitive K-Cl cotransport occurs in red blood cells of many species. In intact cells, activation of K-Cl cotransport by swelling requires dephosphorylation of some cell protein, but maximal activity requires the presence of intracellular ATP. We have examined the relation between K-Cl cotransport activity and ATP in ghosts prepared from human red blood cells. K-Cl cotransport activity in swollen ghosts increased by ATP, and the increase requires Mg so that it almost certainly results from the phosphorylation of some membrane component. However, even in ATP-free ghosts residual volume-sensitive K Cl cotransport can be demonstrated. This residual cotransport in ATP-free ghosts is greater in the presence of vanadate, a tyrosyl phosphatase inhibitor, and in ghosts that contain ATP cotransport is reduced by genistein, a tyrosyl kinase inhibitor. Okadaic acid, an inhibitor of serine and threonine phosphatases, inhibits K-Cl cotransport in ghosts as it does in intact cells. Experiments in which ghosts were preexposed to okadaic acid showed that the protein dephosphorylation that permits K-Cl cotransport can proceed to completion before the ghosts are swollen and K transport measured and therefore dephosphorylation is not a response to ghost swelling. In experiments with ATP-free ghosts we found that phosphorylation is not necessary to increase the cotransport rate when shrunken ghosts are swollen, nor is rephosphorylation necessary to decrease the cotransport rate when swollen ghosts are shrunken. Cotransport is greater in swollen than in shrunken ghosts even when the swollen and shrunken ghosts have the same concentration of cytoplasmic solutes. We conclude that, although phosphorylation and dephosphorylation modify the activity of the cotransporter in swollen and in shrunken ghosts, neither of these processes nor any other known messenger is involved in signal transduction between the cell volume sensor and the cotransporter as originally proposed by Jennings and Al-Rohil (Jennings, M. L., and N. Al-Rohil. 1990. Journal of General Physiology. 95: 1021-1040). PMID- 8245824 TI - Effects of acidosis on resting cytosolic and mitochondrial Ca2+ in mammalian myocardium. AB - Acidosis increases resting cytosolic [Ca2+], (Cai) of myocardial preparations; however, neither the Ca2+ sources for the increase in Cai nor the effect of acidosis on mitochondrial free [Ca2+], (Cam) have been characterized. In this study cytosolic pH (pHi) was monitored in adult rat left ventricular myocytes loaded with the acetoxymethyl ester (AM form) of SNARF-1. A stable decrease in the pHi of 0.52 +/- 0.05 U (n = 16) was obtained by switching from a bicarbonate buffer equilibrated with 5% CO2 to a buffer equilibrated with 20% CO2. Electrical stimulation at either 0.5 or 1.5 Hz had no effect on pHi in 5% CO2, nor did it affect the magnitude of pHi decrease in response to hypercarbic acidosis. Cai was measured in myocytes loaded with indo-1/free acid and Cam was monitored in cells loaded with indo-1/AM after quenching cytosolic indo-1 fluorescence with MnCl2. In quiescent intact myocytes bathed in 1.5 mM [Ca2+], hypercarbia increased Cai from 130 +/- 5 to 221 +/- 13 nM. However, when acidosis was effected in electrically stimulated myocytes, diastolic Cai increased more than resting Cai in quiescent myocytes, and during pacing at 1.5 Hz diastolic Cai was higher (285 +/- 17 nM) than at 0.5 Hz (245 +/- 18 nM; P < 0.05). The magnitude of Cai increase in quiescent myocytes was not affected either by sarcoplasmic reticulum (SR) Ca2+ depletion with ryanodine or by SR Ca2+ depletion and concomitant superfusion with a Ca(2+)-free buffer. In unstimulated intact myocytes hypercarbia increased Cam from 95 +/- 12 to 147 +/- 19 nM and this response was not modified either by ryanodine and a Ca(2+)-free buffer or by 50 microM ruthenium red in order to block the mitochondrial uniporter. In mitochondrial suspensions loaded either with BCECF/AM or indo-1/AM, acidosis produced by lactic acid addition decreased both intra- and extramitochondrial pH and increased Cam. Studies of mitochondrial suspensions bathed in indo-1/free acid-containing solution showed an increase in extramitochondrial Ca2+ after the addition of lactic acid. Thus, in quiescent myocytes, cytoplasmic and intramitochondrial buffers, rather than transsarcolemmal Ca2+ influx or SR Ca2+ release, are the likely Ca2+ sources for the increase in Cai and Cam, respectively; additionally, Ca2+ efflux from the mitochondria may contribute to the raise in Cai. In contrast, in response to acidosis, diastolic Cai in electrically stimulated myocytes increases more than resting Cai in quiescent cells; this suggests that during pacing, net cell Ca2+ gain contributes to enhance diastolic Cai. PMID- 8245825 TI - Microbial cleavage of nitrate esters: defusing the environment. PMID- 8245826 TI - A novel pathway for the catabolism of 4-nitrotoluene by Pseudomonas. AB - Eleven strains of Pseudomonas were isolated by selective enrichment on 4 nitrotoluene (4NT). They all utilized 4NT, 4-nitrobenzyl alcohol (4NBA) or 4 nitrobenzoate (4NBZate) as sole sources of carbon and nitrogen. One strain, TW3, was used for more detailed studies. 4NT-grown cells of TW3 take up O2 when incubated in the presence of 4NBA, 4-nitrobenzaldehyde (4NBZ) and 4NBZate. HPLC analysis of culture supernatants showed that 4NBZ and 4NBZate were formed when 4NT-grown cells wer incubated with 4NBA, whereas only 4NBZate was found when they were incubated with 4NBZ. Two dehydrogenases were detected in extracts of 4NT grown cells. 4NBA dehydrogenase could be assayed by a dye-linked assay whereas 4NBZ dehydrogenase activity was linked to NAD+ reduction. No nitrite was detected in supernatants of 4NBZate-grown cells incubated with 4NBZate but the nitrogen appeared as ammonium. The only aromatic ring-cleavage dioxygenase that was induced during growth on the nitroaromatics was protocatechuate 3,4-dioxygenase. It is proposed that the pathway for 4NT catabolism proceeds via 4NBA, 4NBZ and 4NBZate and ultimately to protocatechuate with release of the nitro group as ammonium. PMID- 8245827 TI - Purification and characterization of plantaricin A, a Lactobacillus plantarum bacteriocin whose activity depends on the action of two peptides. AB - A Lactobacillus plantarum bacteriocin, plantaricin A, has been purified to homogeneity by ammonium sulphate precipitation, binding to cation exchanger and Octyl-Sepharose, and reverse-phase chromatography. The bacteriocin activity was associated with two peptides, termed alpha and beta, which were separated upon reverse-phase chromatography. Bacteriocin activity required the complementary action of both the alpha and beta peptides. From the N-terminal end, 21 and 22 amino acid residues of alpha and beta, respectively, were sequenced. Further attempts at sequencing revealed no additional amino acid residues, suggesting that either the C terminus had been reached or that modifications in the next amino acid residue blocked the sequencing reaction. Judging from their amino acid sequence, alpha and beta may be encoded by the same gene, since alpha appeared to be a truncated form of beta. Alanine, the first amino acid residue at the N terminal end of beta was not present at this position in alpha. Otherwise the sequences of alpha and beta appeared to be identical. The calculated molecular masses of the sequenced part of alpha and beta were 2426 and 2497 Da, respectively. The molecular masses of alpha and beta as determined by mass spectroscopy were 2687 +/- 30 and 2758 +/- 30 Da, respectively, indicating that (i) the only difference between alpha and beta was the presence of the N-terminal alanine residue in beta, and that (ii) in addition to the sequenced residues, two to three unidentified amino acid residues are present at the C-terminal ends of the alpha and beta peptides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245828 TI - Sequence and expression of the Mycobacterium leprae dnaJ gene. AB - Study of Mycobacterium leprae, the causative agent of leprosy, has been advanced by the isolation of genes encoding mycobacterial proteins including dnaK encoding the M. leprae 70 kDa heat shock protein. The sequence downstream from dnaK revealed a second open reading frame coding for a protein of 389 amino acids with a calculated molecular mass of 41.2 kDa. Sequence analysis demonstrated significant DNA homology with the dnaJ gene of other organisms. High amino acid sequence identity was obtained between the DnaJ protein of M. leprae and M. tuberculosis (89%) with significant divergence between the two occurring only at the C-terminal end. The expressed recombinant DnaJ protein had a molecular mass of 42 kDa. PMID- 8245829 TI - A new experimental approach to the search for chemical density factors in the regulation of monoculture growth. AB - In monocultures of micro-organisms, growth is controlled by feedback mechanisms involving chemical factors such as limiting substrates and inhibitory metabolic products. The role of such feedback in the growth regulation of Escherichia coli O-124 was investigated by growing cells in batch culture using a medium containing glucose and mineral salts. In various phases of growth, portions of the native culture were diluted with culture filtrate, so that although cell density decreased, the chemical composition of the growth medium was unaltered. As the diluted cultures grew, variations in growth acceleration were calculated and compared with those of native (undiluted) cultures. Towards the end of the exponential phase and in the growth deceleration phase, the specific feedback level (FBL) was between -20 and -200 (h g l-1)-1. The feedback components resulting from changes in glucose concentrations were calculated using experimentally determined values of mu max (0.55 +/- 0.05 h-1) and Ks (2.5 +/- 0.7 mg l-1). Only 0.1-40% of FBL could be accounted for by changes in glucose concentration, indicating the presence of additional growth regulators. The method developed may become a new tool for determination of growth-regulating cell-density factors in microbial cultures. PMID- 8245830 TI - Temporal activation of beta-glucanase synthesis in Bacillus subtilis is mediated by the GTP pool. AB - beta-Glucanase synthesis was temporally activated in Bacillus subtilis at the onset of stationary phase. This regulation was dependent upon a drop in the GTP concentration in response to nutrient limitation. The Spo0A and AbrB proteins were involved in the GTP-dependent temporal activation. PMID- 8245831 TI - Catabolite repression of beta-glucanase synthesis in Bacillus subtilis. AB - beta-Glucanase synthesis in Bacillus subtilis was repressed by glucose and other substrates of glycolysis. Experiments with different pts mutants showed that the phosphoenolpyruvate: sugar phosphotransferase system is not involved in carbon catabolite repression of beta-glucanase synthesis. Carbon catabolite repression of beta-glucanase synthesis was completely abolished in a ccpA mutant. An operator structure similar to those upstream of amyE and the xyl operon was found and was shown by site-directed mutagenesis to be the target for carbon catabolite repression of beta-glucanase synthesis. The presence of this operator on a multi copy plasmid resulted in a reduced repression of both beta-glucanase and alpha amylase synthesis. It seems likely that the gene encoding these enzymes are part of one regulon with respect to catabolite repression. PMID- 8245832 TI - The giant linear plasmid pHG207 from Rhodococcus sp. encoding hydrogen autotrophy: characterization of the plasmid and its termini. AB - As described previously, in Rhodococcus sp. (formerly Nocardia opaca) strains MR11 and MR22, the ability to grow as an aerobic hydrogen bacterium (the Aut character) is located on giant conjugative linear plasmids--on pHG201 (270 kb) in strain MR11 and on pHG205 (280 kb) in strain MR22. In an autotrophic transconjugant originating from MR22 a smaller plasmid, pHG207 (225 kb), was detected and shown to be a recombination product of the wild-type plasmids pHG204 and pHG205. A donor carrying pHG207 as the sole plasmid transferred the Aut marker at a 1000-fold frequency compared to the wild-type plasmid pHG205. Analysis of the plasmid ends revealed that plasmid pHG207 carries proteins at both ends; the proteins are linked to the 5' ends of the strands. The cloned end fragments of about 2 kb were sequenced and found to contain highly homologous sequences within the terminal 583 bp (left end part) and 560 bp (right end part). Several potential reading frames were detected, but database searching gave no indication about possible functions. PMID- 8245833 TI - Conserved and variable regions in protein Arp, the IgA receptor of Streptococcus pyogenes. AB - The streptococcal M protein family, a number of cell surface molecules that interact with the human immune system, can be divided into two major classes, A and C, characterized by different types of repeats in the central part of the molecule. Class A and class C molecules are known to have a variable N-terminal region and a more conserved C-terminal region, but little is known about the mechanisms that give rise to this structural variation. In this report, we show that two variants of protein Arp, an IgA receptor in class C of the M protein family, have virtually identical signal sequences and C-terminal halves, but unrelated N-terminal sequences. Comparison of the sequences of the two genes and their flanking regions also demonstrates the presence of well-defined variable and conserved regions. Our results strongly suggest that the N-terminal sequence variation between the two variants of protein Arp was generated through an intergenic recombination event, rather than through intragenic recombination or accumulation of mutations. PMID- 8245834 TI - Random amplified polymorphic DNA markers reveal a high degree of genetic diversity in the entomopathogenic fungus Metarhizium anisopliae var. anisopliae. AB - Metarhizium anisopliae isolates from several insect hosts and from various sugar cane growing areas of Queensland, Australia, were examined for genetic diversity using random amplified polymorphic DNA (RAPD) markers. Thirty isolates of M. anisopliae var. anisopliae and one isolate of M. anisopliae var. majus were examined. Ten randomly chosen 10mer or 11mer primers were used and RAPD banding patterns were compared. Thirty distinct genotypes could be distinguished amongst the 31 isolates tested on the basis of RAPD patterns. Six of the isolates classified as M. anisopliae var. anisopliae exhibited closer similarity to the M. anisopliae var. majus isolate than to other anisopliae strains tested. Isolates exhibiting similar (> 80% similarity) RAPD profiles tended to be isolated from the same geographic area and evidence for the persistence of particular fungal genotypes in specific geographical localities was obtained. Pathogenicity assays suggested that, in some instances, RAPD groupings may also indicate insect host range. The mean similarity amongst isolates measured by band sharing in all pairwise comparisons was 41% and the most distinct pair of isolates shared only 9% of their RAPD bands. We conclude that the isolates tested belonging to the species M. anisopliae, as assessed on morphological grounds, represent a very diverse genetic group. The results also suggest that RAPD markers may be useful for the tracking of specific biocontrol strains in the field. PMID- 8245835 TI - The Sc7/Sc14 gene family of Schizophyllum commune codes for extracellular proteins specifically expressed during fruit-body formation. AB - The Sc7 and Sc14 genes are specifically expressed in the dikaryon of the basidiomycete fungus Schizophyllum commune during fruiting. These genes are closely linked (within 6 kb) and highly similar in gene structure and nucleotide sequence (70% identical nucleotides in their coding regions). The encoded proteins (204 and 214 amino acids, respectively) have 87% similarity in amino acids (56% of the amino acids are identical). They contain putative signal sequences for secretion, are rich in aromatic amino acids which are generally located at similar positions, and they are generally hydrophilic. Inspection of databanks showed similarities with pathogenesis-related proteins (PR1) from plants, testis-specific proteins from mammals and venom allergen proteins from insects. An antibody raised against a Sc7 fusion protein showed the presence of the Sc7 protein in the culture medium and in the fruit bodies where it is apparently loosely associated with hyphal walls. PMID- 8245836 TI - Inactivation of the CDC25 gene product in Saccharomyces cerevisiae leads to a decrease in glycolytic activity which is independent of cAMP levels. AB - In the budding yeast Saccharomyces cerevisiae cyclic AMP (cAMP) can influence the activity of key enzymes in carbohydrate metabolism through modulation of the activity of cAMP-dependent protein kinase. One of the components involved in cAMP production is the CDC25 gene product, which can activate the RAS/adenylate cyclase pathway by promoting the exchange of guanine nucleotides bound to RAS. In two yeast strains carrying different thermosensitive alleles of the CDC25 gene, cAMP levels respond differently to an increase in growth temperature from 23 degrees C (permissive) to 36 degrees C (restrictive). In strain OL86 (cdc25-5) the estimated intracellular concentration of cAMP dropped after transfer to restrictive temperature whereas in strain ts321 (cdc25-1) the cAMP level rose under the same conditions. Despite the differences in cAMP levels the glycolytic flux in the two mutants responded in a very similar way to the shift from permissive to restrictive temperature; after the increase in the incubation temperature, the specific glycolytic flux in both cdc25-1 and cdc25-5 initially increased from about 300 nmol min-1 (mg protein)-1 to about 500 nmol min-1 (mg protein)-1 (presumably mainly as a consequence of the increase in temperature), but then gradually fell to 100-200 nmol min-1 (mg protein)-1. A similar pattern of CO2 production to that found in the two cdc25 mutants was also observed for several other thermosensitive mutants displaying a Start-II type of G1 arrest. In contrast, in a wild-type strain and in strains giving a Start-I type of G1 arrest, CO2 production did not drop after a temperature shift. The specific activities of glycolytic enzymes in the two cdc25 mutants did not show much change after the temperature shift, indicating that the decrease in glycolytic flux was not caused by a decrease in the activity of any of the glycolytic enzymes. Our data show that, at least in long-term regulation, the cAMP levels per se are not likely to be a prime factor controlling glycolytic flux. PMID- 8245837 TI - Influence of medium components and metabolic inhibitors on citric acid production by Penicillium simplicissimum. AB - Penicillium simplicissimum excreted more than 100 mmol citric acid l-1 [2.9 mmol (g dry wt)-1; 9 d] if an industrial filter dust (> 50% ZnO) providing a high extracellular buffering capacity was present in the medium. A similar specific [2 mmol (g dry wt)-1], but lower absolute (26 mmol l-1), citric acid excretion occurred in the absence of an extracellular buffer and if amino acids or urea were used as nitrogen source. P. simplicissimum excreted no citric acid under conditions where Aspergillus niger produces citric acid (deficiency of trace elements, low pH and reduced biomass formation). Citric acid excretion by P. simplicissimum always paralleled biomass formation and occurred in a pH range between 4 and 7. This indicated that different imbalances of metabolism were responsible for citric acid excretion in A. niger and P. simplicissimum. However, provided a high extracellular buffering capacity was present, the response of the Penicillium system to different carbon and nitrogen sources was similar to the Aspergillus system. In contrast, the metals iron and copper had virtually no effect on citric acid excretion compared with A. niger. Estimation of intracellular citric acid, as well as the effects of the uncoupler 2,4 dinitrophenol, and the H(+)-ATPase inhibitor sodium orthovanadate, led to the conclusion that the buffer-stimulated citric acid efflux was dependent on metabolic energy and an energized plasma membrane, respectively. Despite similarities to the Aspergillus system, a different mechanism for buffer stimulated citric acid excretion by P. simplicissimum seems probable. PMID- 8245838 TI - The cell wall of the oleaginous yeast Trichosporon cutaneum. AB - The cell wall of Trichosporon cutaneum consists of 11% protein, 63% neutral carbohydrate, 9% glucosamine and 13% glucuronic acid. The sugars include glucose (32%), mannose (6%) and traces of xylose and galactose. The cell wall was fractionated with alkali to yield a mixture of alkali-soluble matrix components, and an alkali-insoluble glucan associated with chitin. The alkali-insoluble glucan contained a mixture of (1-3) and (1-6) glycosidic linkages. It was only partly susceptible to digestion by the beta(1-3) glucanase, Zymolyase. The alkali soluble fraction contained glucan, mannan and acidic polymers. The glucan was (1 3)-linked with no (1-6) linkages and only trace amounts of (1-3-6)-linked glucose. It was resistant to digestion by Zymolyase. Extensive hydrolysis of this fraction with trifluoroacetic acid released a high-molecular-mass glucuronan which had 1H- and 13C-NMR profiles matching those of the beta(1-4) glucuronan, mucoric acid. Xylomannan was purified from isolated cell walls and from whole cells. It contained glucose, mannose, xylose, and D-glucuronic acid. It was very similar in composition and structure to the capsular polysaccharides of Cryptococcus neoformans, and to an extracellular polysaccharide produced by another yeast described as T. cutaneum. Electron microscopy showed that the cell wall of T. cutaneum has a lamellar structure characteristic of a basidiomycetous yeast rather than the electron-dense 'fuzzy coat' seen in Candida albicans. PMID- 8245839 TI - Molecular analysis of a gene encoding a serum-resistance-associated 76 kDa surface antigen of Haemophilus somnus. AB - Haemophilus somnus is a Gram-negative bacterial bovine pathogen which can cause disease or be carried asymptomatically. We previously showed that four serum sensitive isolates from asymptomatic carriers lacked a 13.4 kb sequence of chromosomal DNA that was present in two virulent serum-resistant strains. We have since sequenced 5 kb of the 13.4 kb fragment from a serum-resistant strain, which contained an open reading frame (ORF) of at least 4.5 kb. From Western blot analysis, the ORF was shown to encode a 76 kDa protein (p76) that co-migrated with a 76 kDa H. somnus surface protein. Both the recombinant and natural p76 reacted with convalescent-phase serum from a cow in an experimental H. somnus abortion study. The translational start site for p76 was identified by deletion analysis of subclones of the 5 kb cloned sequence. The 4.5 kb ORF contained 1.2 kb tandem direct repeats (DRs), with 65% identity between the two repeats at the protein level. The 5' DR (DR1) included the start site for the 76 kDa protein, and DR2 had a flanking inverted repeat, suggestive of an insertion-sequence-like element. PMID- 8245840 TI - Chlamydia trachomatis infection of cultured motile cells after uptake of chlamydiae from the substratum. AB - The ability of motile cells to remove small inanimate particles from solid substrata is well documented. We show here that motile cells will pick up and internalize infectious particles of the obligate intracellular parasite Chlamydia trachomatis when they are adherent to the substratum over which the host cells move. Two cell types were used to assess chlamydial uptake; a feeder independent human squamous cell carcinoma variant (AC3A cells) and the McCoy cell line. Purified chlamydia elementary bodies were attached to glass or collagen-coated glass by centrifugation. Suspensions of cells were then allowed to sediment on to the substrata to which chlamydiae had attached. Both types of cell picked up chlamydiae and transported them over their surface during the course of attachment and spreading. Stereoscopic images obtained by confocal microscopy demonstrated that chlamydiae were found mainly on the surface of non-spread cells. After the cells had spread on the substratum they began to move around forming tracks where the chlamydiae had been removed. Some cell-surface-attached chlamydiae were endocytosed and a proportion of these proliferated during the 48 h after plating. However, chlamydiae attached to the substratum lost infectivity by a simple exponential decay process within a few hours of incubation in the extracellular environment. Therefore, increasing numbers of non-viable organisms were probably endocytosed as the time of extracellular incubation increased. This mode of infection may be relevant to in vivo situations where cell migration occurs after damage to mucosal surfaces. PMID- 8245841 TI - Purification of yersiniabactin: a siderophore and possible virulence factor of Yersinia enterocolitica. AB - HPLC analysis revealed that Yersinia enterocolitica WA-C produced two substances under iron-limiting conditions one of which was identified as 2,3 dihydroxybenzoyl-L-serine. The other compound had iron-complexing activity and was called yersiniabactin. The fur mutant H1852 was shown to produce yersiniabactin constitutively in an iron-independent manner. Yersiniabactin was isolated by ethyl acetate extraction from the spent medium of H1852, size fractionation chromatography and preparative HPLC. A catechol function was demonstrated with different chemical assays and by UV-visible spectroscopy. The molecular mass of yersiniabactin was determined to be 482 Da. Purified yersiniabactin stimulated growth of Y. enterocolitica and Escherichia coli phi under iron-limiting conditions and apparently served as an iron carrier. Transport of 55Fe-yersiniabactin was TonB-dependent, indicating a receptor mediated uptake across the outer membrane. A pesticin-resistant mutant missing the receptor protein FyuA was unable to transport and use yersiniabactin as a siderophore. PMID- 8245842 TI - Biological activities of cell envelope fragments of the archaeobacterium Sulfolobus solfataricus: lethal toxicity, local hypersensitivity, pyrogenicity and spleen lymphocyte mitogenicity. AB - The sensitizing effect and the local and general toxicity related to membrane components of the archaeobacterium Sulfolobus solfataricus was studied. Cell envelope fragments were biologically active but this activity was lost upon separation of the lipid and protein components. The envelope fragments exerted lethal effects on mice sensitized with D-galactosamine that were prevented by pretreatment with anti-TNF-alpha serum. This lethal activity occurred in both LPS responder (BALB/cByJ) and LPS-nonresponder (C3H/HeJ) mouse strains. In addition, Sulfolobus envelope fragments tested in rabbits caused a local Schwartzman reaction, and showed pyrogenic activity. In vitro, the envelope fragments that act on spleen lymphocytes of the LPS-responder (BALB/cByJ) and LPS-nonresponder (C3H/HeJ) mice caused an uptake of [3H]thymidine similar to that caused by concanavalin A. A similar toxic activity to that exerted by eubacteria is therefore exerted by this non-pathogenic archaeobacterium despite the difference in surface chemistry. PMID- 8245843 TI - A haemagglutinating adhesin of group B streptococci isolated from cases of bovine mastitis mediates adherence to HeLa cells. AB - Rabbit erythrocytes were agglutinated by 43.4% of group B streptococci isolated from bovines but by none isolated from humans. Haemagglutination was enhanced by cultivation of the bacteria under microaerophilic conditions. Most of the haemagglutinating strains had protein type antigen X, either alone, or in combination with polysaccharide antigens. Heat and proteolytic treatment of the bacteria destroyed the haemagglutination activity. The haemagglutinin could be solubilized from the bacterial surface by mutanolysin treatment and isolated from culture supernatant fluid by ammonium sulphate precipitation. The isolated haemagglutinin did not cause direct agglutination of erythrocytes. However, binding of the haemagglutinin to rabbit erythrocytes could be visualized by agglutination of haemagglutinin-treated erythrocytes by specific antiserum obtained by absorption. Western blotting showed that the haemagglutinin obtained from erythrocyte lysates contained an antibody-reactive band with a molecular mass of 43 kDa. Haemagglutination-positive strains adhered to HeLa cells in higher numbers than did haemagglutination-negative strains. The HeLa cell adherence of Group B streptococci was inhibited in the presence of isolated haemagglutinin or of specific antiserum against the haemagglutinin. These observations suggest that the haemagglutinating adhesins of bovine group B streptococcal isolates are directly involved in the adherence mechanisms of these organisms. PMID- 8245844 TI - Analysis of intra-specific variation in the fatty acid profiles of Borrelia burgdorferi. AB - Analysis of the fatty acid methyl esters (FAMEs) of bacteria is a commonly used chemotaxonomic technique. Application of this methodology to spirochaetes associated with Lyme borreliosis revealed distinct clusters corresponding to three genetically distinguished groups: Borrelia burgdorferi sensu stricto, B. garinii, and the VS461 group. However, B. garinii formed a common group with B. hermsii, a relapsing fever spirochaete, and VS461 grouped with B. turicatae and B. parkeri, two other relapsing fever spirochaetes. The diversity in fatty acid profiles of Lyme disease spirochaetes has implications for the protean clinical manifestations of the disease. PMID- 8245845 TI - A simple chemical test to distinguish mycobacteria from other mycolic-acid containing actinomycetes. AB - Two hundred and fifty-two representatives of the general Corynebacterium, Gordona, Mycobacterium, Nocardia, Rhodococcus and Tsukamurella were degraded by alkaline hydrolysis and their mycolic acids extracted as methyl esters following phase-transfer-catalysed esterification. When the mycolic acid methyl esters were treated with a mixture of acetonitrile and toluene all mycobacterial mycolates formed copious white precipitates whereas all but 5 out of the 106 non mycobacterial mycolates remained in solution. The precipitated methyl mycolates and the dried soluble mycolates were compared by pyrolysis gas chromatography and silica gel thin-layer chromatography. On pyrolysis, the precipitated methyl mycolates from mycobacteria yielded fatty acid methyl esters with 20 to 26 carbon atoms whereas those from the remaining taxa produced shorter-chain esters. Mycobacteria and Tsukamurella paurometabola gave multispot mycolic acid patterns on thin-layer chromatography of their methyl esters whereas those from the remaining strains gave single spots. Our results indicate that Rhodococcus chlorophenolicus strains contain mycolic acids atypical of mycobacteria. It can be concluded that the mycolic acid precipitation test provides a simple and reliable way of distinguishing mycobacteria from all other prokaryotes, notably from other mycolic-acid-containing taxa. PMID- 8245846 TI - Antibody and DNA probes for detection of nitrite reductase in seawater. AB - A polyclonal antiserum was produced by immunization with nitrite reductase (NiR) purified from Pseudomonas stutzeri (ATCC 14405) and tested for specificity among known denitrifying strains. The antiserum was nearly strain-specific, identifying NiR only in some, but not all, other P. stutzeri strains. Denitrifying isolates from water column and sediment environments were also screened; several isolates from an intertidal microbial mat reacted with the NiR antiserum. Activity assays for NiR in polyacrylamide gels demonstrated that strains with apparently very similar NiR proteins did not react with the antiserum. These results imply that the NiR protein is more variable even among closely related strains than previously suspected. A DNA probe for a 721 bp region of the NiR structural gene was obtained by PCR amplification of P. stutzeri (ATCC 14405) DNA and used to screen denitrifying strains and isolates. The probe hybridized with a greater variety of strains than did the antiserum, implying that the DNA probe may be a more broadly useful and functional probe in environmental samples, whilst the NiR antiserum is nearly strain- or, at most, species-specific. Limits for detection of the enzyme and gene in seawater were estimated and NiR DNA was detected in DNA extracted from natural seawater. The hybridization data imply that in the order of 1-10 in 1000 cells in natural seawater possess homology with the NiR gene probe. PMID- 8245847 TI - Toroviruses: replication, evolution and comparison with other members of the coronavirus-like superfamily. PMID- 8245848 TI - Mapping and sequence of the gene encoding the African swine fever virion protein of M(r) 11500. AB - The gene encoding the African swine fever virus protein of M(r) 11,500, present in the virus particle, has been mapped and sequenced in the genome of the Vero cell-adapted virus strain BA71V. A serum raised against virion proteins of M(r) 12,000 to 13,000 isolated from polyacrylamide gels was used to screen a plasmid expression library, containing viral DNA random fragments, that expresses viral polypeptides fused to beta-galactosidase. Using this method, we have identified and sequenced the open reading frame (ORF) A137R, which initiates at the right end of the EcoRI A restriction fragment and extends into the EcoRI F fragment. Expression of the protein in Escherichia coli has confirmed that ORF A137R encodes a protein with an M(r) of about 12,000. A specific serum was raised against the E. coli-expressed protein, and has been used to identify the protein encoded by the ORF, which is translated at late times of infection and incorporated into the virus particle. Immunofluorescence experiments have shown that the protein localizes in virus factories. PMID- 8245850 TI - Tumour necrosis factor alpha stimulates the activity of the human cytomegalovirus major immediate early enhancer/promoter in immature monocytic cells. AB - Both tumour necrosis factor alpha (TNF-alpha) and phorbol 12-myristate 13-acetate (PMA) stimulated human cytomegalovirus (HCMV) major immediate early (IE) enhancer/promoter activity in the HL-60 granulocyte/monocyte progenitor cell line when added to transfected cells. In U-937 monocytic cells, by contrast, TNF-alpha had no stimulatory effect and the addition of PMA produced only marginal stimulation. In the mature THP-1 monocytic cell line and in differentiated HL-60 cells, addition of TNF-alpha caused inhibition of the IE enhancer/promoter activity. The stimulating effect of PMA, as observed in the other cell lines, however, remained. Thus the effect of TNF-alpha on the major IE enhancer/promoter activity is determined by the degree of differentiation of the infected cells. Unlike TNF-alpha and PMA, the interleukins IL-1, IL-3, IL-6 as well as the cytokine GM-CSF were found to have no detectable influence on the activity of the IE enhancer/promoter activity which, likewise, was not affected by the presence of the modulator sequence. Since premonocytic cells are suggested to be sites of HCMV latency, the stimulation by TNF-alpha could be of potential pathophysiological significance. PMID- 8245849 TI - Mapping the genetic region coding for herpes simplex virus resistance to mouse interferon alpha/beta. AB - Herpes simplex virus (HSV) ocular virulence has been associated with strain sensitivity to mouse interferon (IFN)-alpha/beta. To identify the region of the virus genome associated with heightened resistance to this cytokine, intertypic recombinants were constructed using the intact genome of avirulent, IFN-sensitive HSV type 1 (strain 35) and XbaI-digested DNA from virulent, IFN-resistant HSV type 2 (strain 186). An intertypic recombinant, designated HSV-R4, was isolated which grew to titres 10- to 100-fold higher than HSV-1(35) in mouse ocular tissue in vivo, and induced stromal keratitis. The recombinant which was several orders of magnitude more resistant to mouse IFN-alpha/beta than HSV-1(35) had a genome composed of HSV-1(35) DNA except for a 12 kb fragment (0.15 to 0.23 map units) derived from HSV-2(186). To define the IFN resistance locus further, three overlapping subclones of this 12 kb fragment were constructed from the HSV-2(186) genome and subjected to marker rescue experiments. The cloned BamHI D fragment was the only subclone that promoted HSV-1(35) ocular growth in vivo. An intertypic recombinant, designated HSV-R(BD), was isolated from the 35 x 186 BamHI D transfection progeny pool. This recombinant, in contrast to HSV-1(35), was several orders of magnitude more resistant to mouse IFN-alpha/beta inhibition in vitro, grew 10- to 100-fold better in mouse ocular tissue in vivo, and caused severe necrotizing stromal keratitis in BALB/c mice. Analysis of the recombinant genome indicated that the HSV-2 genetic information responsible for IFN resistance of HSV-R(BD) was located within the BamHI D fragment, most likely mapping to that region containing three partial open reading frames designated UL14, UL15 and UL16. The products encoded by this region remain to be identified. PMID- 8245851 TI - Interactions between equine herpesvirus type 1 and equine herpesvirus type 4: T cell responses in a murine infection model. AB - Interactions involving the immune responses to equine herpesvirus types 1 and 4 (EHV-1 and EHV-4) were studied in a murine infection model. When mice were inoculated intranasally with EHV-1, virus replication occurred in the respiratory tract and clinical signs were produced. In contrast, mice that were similarly inoculated with EHV-4 produced no evidence of virus replication and showed no clinical signs. When mice that had been inoculated with live EHV-4 were challenged 1 month later with EHV-1 they were partially protected. Although clinical signs were apparent on reinfection, virus replication in the respiratory tract was reduced in these mice compared with control mice that had not been previously immunized. Mice primed with heat-inactivated EHV-4, however, were not so protected. Live EHV-4-primed mice developed very low levels of antibody to EHV 1 and the humoral response could not account for this protection. However, the infected mice did give a strong delayed-type hypersensitivity reaction in a skin test using either EHV-1 or EHV-4 antigen. Spleen cells from EHV-4-primed donors provided a source of immune cells, including T cells which were used for transfer to recipient mice which were then challenged with EHV-1. The cells were protective; there was a reduction of virus replication on challenge with EHV-1 which correlated with the number of cells transferred. Modulation of the protective effect of primed cell populations was tested after depletion in vivo by means of complement-mediated lysis. The depletion of CD4-bearing cells produced the least effect on the protection afforded by cell transfer. In contrast, depletion of CD8-bearing cells markedly reduced the protection in recipients. EHV-1 and EHV-4 are widespread in horses and cross-infections are common. These results gained from a murine model indicate that important interactions occur at the level of T cell immunity between the two virus types which warrant further investigation in the natural host. PMID- 8245852 TI - Measles virus glycoproteins: studies on the structure and interaction of the haemagglutinin and fusion proteins. AB - We have investigated the structure and interaction of the measles virus (MV) glycoproteins expressed at the cell membrane. Cross-linking studies with a variety of chemicals stabilized dimeric forms of the haemagglutinin (HA) or fusion (F) proteins, although by sucrose density gradient analysis, oligomers corresponding to tetramers and larger were observed for both proteins. In cells in which both HA and F were expressed at the surface, their close association was shown by cross-linking and co-immunoprecipitation. PMID- 8245853 TI - Expression of the S1 and S2 subunits of murine coronavirus JHMV spike protein by a vaccinia virus transient expression system. AB - The spike (S) protein of murine coronavirus JHMV, variant cl-2, comprises two polypeptides, N-terminal S1 (with an N-terminal signal peptide) and C-terminal S2 (with a C-terminal transmembrane domain). In order to express these subunits, we constructed three different vaccinia virus transfer vectors (VV-TVs) containing cDNAs encoding the S1 protein without a transmembrane domain (pSFS1utt), the S1 protein with a C-terminal transmembrane domain derived from S2 (pSFS1tmd) or the S2 protein with an N-terminal signal peptide derived from S1 (pSFssS2). The S1 and S2 proteins were expressed in DBT cells by infection with vaccinia virus and transfection of these VV-TVs. In cells transfected with the pSFS1utt and pSFS1tmd, 96K and 106K proteins, respectively, were detected by Western blotting. The ssS2 protein expressed by pSFssS2 was 96K, which was slightly larger than the authentic S2 protein. The S1utt and S1tmd proteins were shown by binding studies using a panel of monoclonal antibodies to be antigenically indistinguishable from the authentic S1 protein. The S1tmd and ssS2 proteins were detected on the cell surface by immunofluorescence, whereas the S1utt protein was not. However, when the S1utt protein was expressed together with the ssS2 protein, the S1utt was detected on the cell membrane. This suggested that the S1utt was associated with ssS2 on the cell membrane. These observations indicate that the expressed S1 and S2 proteins associated in a similar manner to the authentic S1 and S2 proteins produced in DBT cells infected with cl-2. However, cell fusion was not observed in cells expressing either S1 or S2 nor in cells coexpressing both S1 and S2, although the whole S protein expressed by VV-TV did induce fusion. PMID- 8245854 TI - Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region. AB - Hepatitis C virus (HCV) showed substantial nucleotide sequence diversity distributed throughout the viral genome, with many variants showing only 68 to 79% overall sequence similarity to one another. Phylogenetic analysis of nucleotide sequences derived from part of the gene encoding a non-structural protein (NS-5) has provided evidence for six major genotypes of HCV amongst a worldwide collection of 76 samples from HCV-infected blood donors and patients with chronic hepatitis. Many of these HCV types comprised a number of more closely related subtypes, leading to a current total of 11 genetically distinct viral populations. Phylogenetic analysis of other regions of the viral genome produced relationships between published sequences equivalent to those found in NS-5, apart from the more highly conserved 5' non-coding region in which only the six major HCV types, but not subtypes, could be differentiated. A new nomenclature for HCV variants is proposed in this communication that reflects the two-tiered nature of sequence differences between different viral isolates. The scheme classifies all known HCV variants to date, and describes criteria that would enable new variants to be assigned within the classification as they are discovered. PMID- 8245855 TI - Hexamethylene bisacetamide activates the human immunodeficiency virus type 1 provirus by an NF-kappa B-independent mechanism. AB - Expression of the human immunodeficiency virus type 1 (HIV-1) provirus in T lymphocytic and monocytic cells can be induced by treatment with hexamethylene bisacetamide (HMBA). The induction occurs at the transcriptional level within 1 to 3 h after the addition of the drug, and is not associated with detectable changes in the binding of transcription factors to the enhancer, TATA box or other regulatory regions of the HIV-1 long terminal repeat (LTR). Using the 5' deletion mutants of HIV-1 LTR controlling the expression of the chloramphenicol acetyltransferase gene, we found that the deletion of the kappa B enhancer did not affect HIV-1 inducibility, whereas the deletion of the Sp1 binding sites abolished transcriptional activation. However, the presence of the HIV-1 LTR Sp1 binding sites in the context of the heterologous promoter did not induce responsiveness to HMBA. We conclude that HMBA increases transcription through the secondary modification of the basal transcription complex suggesting the existence of a regulatory pathway that circumvents the requirement for the induction of NF-kappa B or other DNA-specific binding proteins. PMID- 8245856 TI - Feline immunodeficiency virus gene expression: analysis of the RNA splicing pattern and the monocistronic rev mRNA. AB - The transcription pattern of the feline immunodeficiency virus (FIV) genome in a feline CD4+ cell line was examined. In addition to the genomic RNA (9.2 kb), at least five FIV-specific transcripts [5.2, 4.4 (doublet), 1.7 and 1.4 kb] were detected by using subgenomic restriction enzyme fragments of an FIV molecular clone or FIV-specific oligonucleotides as probes. Among these transcripts, the 9.2, 5.2 and 4.4 (doublet) kb mRNAs were not expressed in the cytoplasm of cells transfected with a rev- mutant. To determine the location of splice junctions in the FIV genome, we used PCR to amplify and clone cDNAs corresponding to the viral mRNAs from infected cells. The region between pol and env was found to contain at least two splice donor and three splice acceptor sites. Two splice acceptor sites were detected in the 3' region of env. By hybridization analysis and sequencing of cDNA clones, it was revealed that the medium sized mRNAs are derived from a single splice event, with different splice acceptor sites, and that the two smaller transcripts are doubly or triply spliced mRNAs. Our results demonstrate a complex pattern of alternative splicing of FIV mRNAs. Furthermore, we identified monocistronic rev mRNA species that employ a unique splice acceptor site. PMID- 8245857 TI - Characterization of a new bacteriophage which infects bacteria of the genus Acidiphilium. AB - A novel bacteriophage, termed phi AC1, that infects strains of the genus Acidiphilium (acidophilic, heterotrophic, aerobic, Gram-negative eubacteria) most commonly isolated from acidic mine drainage environments, has been discovered and several of its properties have been determined. This is the first report of a bacteriophage infecting such cells. The virion has a lambdoid morphology and is larger than lambda, as shown by electron microscopy and sucrose gradient centrifugation. The sedimentation coefficient of the virion is approximately 615S. The nucleic acid of phi Ac1 is dsDNA, approximately 102 kb in length. Several experimental results show that phi Ac1 is a temperate phage. The plaques are turbid, and most cells isolated from plaques produced on sensitive cells by filter-sterilized phage preparations contain the phage and are resistant to further phage infection. Southern blot analysis shows that phi Ac1 prophage DNA is integrated into the bacterial genome during the temperature growth phase. PMID- 8245858 TI - Isolates of citrus exocortis viroid recovered by host and tissue selection. AB - Isolates of citrus exocortis viroid (CEV) from a single sweet orange citrus source have been selected by sequential passage through the alternative hosts citron, Gynura aurantiaca, a hybrid tomato Lycopersicon esculentum x L. peruvianum, and from disorganized callus culture of the hybrid tomato. The distinctions in symptom expression, titre and electrophoretic mobility among the CEV isolates, operationally termed CEVc (citron), CEVg (Gynura), CEVt (tomato) and CEVcls (callus) are supported by characteristically different nucleotide sequences. The nucleotide sequence of full-length cDNA clones of CEVc purified from citron shows exchanges not reported for any previously described CEV variant. An unusual number of exchanges have been localized in the terminal domains of all the isolates analysed here. A common pattern of nucleotide exchanges, described as a 'tomato signature', can be detected in all of the isolates derived from hybrid tomato tissues. PMID- 8245859 TI - Nucleotide sequence evidence for the occurrence of three distinct whitefly transmitted geminiviruses in cassava. AB - The complete nucleotide sequence of the DNA of Indian cassava mosaic virus (ICMV) and a key part of that of a group B isolate of African cassava mosaic virus from Malawi (ACMV-M) were determined and compared at the nucleotide and encoded amino acid levels with the published sequences of an ACMV group A isolate (ACMV-K) and other whitefly-transmitted gemini-viruses (WTGs). The DNA of ICMV consists of two circular single-stranded molecules, DNA-A [2815 nucleotides (nt)] and DNA-B (2645 nt), which differ substantially in sequence from the genome components of ACMV-K (DNA-A 70%, DNA-B 47% sequence identity) and other WTGs. ICMV DNA-A contains eight open reading frames (ORFs) encoding proteins of > 100 amino acid residues, of which four ORFs (one genome sense, three complementary sense) are comparable to those of other WTGs. DNA-B contains one ORF in each sense, as in other WTGs. None of the putative viral proteins are more similar in amino acid sequence to the proteins of ACMV-K than to those of another WTG. The coat protein of ACMV-M is more like that of tomato yellow leaf curl virus from Sardinia (86% sequence identity) than those of ICMV or ACMV-K. The intergenic regions of ACMV-K, ACMV-M and ICMV DNAs differ in size, and largely in sequence, except for two 30 to 40 nt sequences which are also conserved in other WTGs and can form stem-loop structures. The intergenic region of ICMV DNA contains three copies of a 41 nt sequence, and that of ACMV-M DNA contains an imperfect repeat of a 34 nt sequence which resembles the repeated sequence in ICMV DNA. The differences between ACMV K, ACMV-M and ICMV are considered great enough to justify their separation as isolates of three distinct WTGs: African cassava mosaic virus, East African cassava mosaic virus and Indian cassava mosaic virus. PMID- 8245860 TI - Interference with brome mosaic virus replication by targeting the minus strand promoter. AB - Sense and antisense strategies for interfering with the replication of brome mosaic virus (BMV) were examined. The effects of 200 nucleotide-long sense and antisense transcripts, corresponding to the viral 3' end (-) strand promoter, on the accumulation of progeny viral RNAs were studied by co-inoculation with wildtype BMV RNAs. Progeny accumulation in barley protoplasts transfected with either sense or antisense transcripts of the (-) strand promoter and BMV RNAs-1 and -2 was decreased by more than 90%, and by 60 to 80% when RNA-3 was also present. This trans interference was concentration-dependent, and reduced both (+) and (-) strand progeny accumulation to a similar extent. The appearance of complementary (-) strands indicated that sense interfering transcripts could serve as templates for (-) strand synthesis, and the use of deletion mutants revealed that the observed interference was in part mediated by this template activity. The reproducibility of the protoplast assay used here allows rapid evaluation of interference strategies and comparisons to be made of alternative approaches to engineered resistance. The results presented here suggest that targeting viral (-) strand promoters with sense and antisense transcripts may be an effective method for engineering plant resistance to viral infection. PMID- 8245861 TI - Localization of a single-stranded RNA-binding domain in the movement protein of red clover necrotic mosaic dianthovirus. AB - Mutant movement proteins of red clover necrotic mosaic dianthovirus (RCNMV), consisting of in-frame deletions or fusions with a maltose-binding protein, were produced in Escherichia coli using expression vectors. The ability of the mutant proteins to bind to ssRNA was tested by photochemical cross-linking and gel retardation. The results showed that the region between amino acids 181 and 225 of the RCNMV movement protein contains an ssRNA-binding domain. PMID- 8245862 TI - Effect of the alfalfa mosaic virus movement protein expressed in transgenic plants on the permeability of plasmodesmata. AB - Symplastic transport of different sized fluorescent probes has been assessed in leaf epidermal cells of transgenic Nicotiana plants expressing the movement protein (MP) of alfalfa mosaic virus (AMV). In both N. tabacum and N. benthamiana, the size exclusion limit (SEL) of plasmodesmata increased from M(r) 1000, which represents the commonly accepted limit, to over 4.4K. However, in control plants, movement of a 3K probe was seen in 11 to 22% of the injections, indicating that plasmodesmata may on occasion allow the passage of molecules larger than was previously thought. The increase of SEL due to the presence of the AMV MP, although significant, remains insufficient to permit the passage of viral particles and the possibility of other mechanisms involved in viral cell-to cell spread is discussed. PMID- 8245863 TI - Complete nucleotide sequence and coding strategy of rice hoja blanca virus RNA4. AB - The complete sequence of rice hoja blanca virus (RHBV) RNA4 has been determined, based on the sequence of the corresponding cDNA clones. RNA4 consists of 1991 nucleotides with two open reading frames (ORFs). One putative ORF is located in the 5'-proximal region of the viral RNA4; it encodes a protein of predicted M(r) 20076 which corresponds to the major non-structural protein that accumulates in RHBV-infected rice plants, and which bears limited sequence identity with the helper component of tobacco vein mottling potyvirus. The other ORF is located in the 5'-proximal region of the viral complementary RNA4 and encodes a protein of predicted M(r) 32,469. Between the two ORFs is an intergenic region of 524 nucleotides, part of which can theoretically adopt a stable stem-loop structure; the 5' and 3' ends can potentially base-pair over 16 nucleotides, producing a pan handle configuration. These characteristics are in favour of an ambisense coding strategy for RHBV RNA4. PMID- 8245864 TI - Reasons for the low accumulation level of aphid transmission factor protein in infected leaves with an aphid-non-transmissible cauliflower mosaic virus isolate, CM1841. AB - The synthesis and accumulation of aphid transmission factor protein (p18) in cauliflower mosaic virus (CaMV)-infected turnip protoplasts were examined in time course and pulse-labelling experiments, comparing an aphid-non-transmissible isolate (CM1841) with an in vitro recombinant aphid-transmissible CaMV (CMBX) generated from the CM1841 isolate. There was little difference in the synthesis and accumulation of p18 between CM1841- and CMBX-infected protoplasts. When the accumulation of p18 in infected leaves was monitored from 3 to 28 days post symptom emergence (p.e.) by Western blotting, the amount of p18 accumulated in CM1841-infected leaves continuously decreased from 3 days p.e. throughout the experimental period, whereas the amount of p18 in CMBX-infected leaves was lowest at 3 days p.e. and increased thereafter. These results suggested that CM1841 differed from CMBX not in the synthesis of p18 but in the stability of p18 in infected leaves. PMID- 8245865 TI - Characterization of RNA-binding domains of hepatitis delta antigen. AB - Hepatitis delta antigen (HDAg), the only protein encoded by the hepatitis delta virus (HDV), binds specifically genomic and antigenomic strands of the HDV RNA. In a previous study, three recombinant HDAg subdomains were synthesized, covering residues 11 to 78, 79 to 163 and 164 to 212, and only the middle domain was shown to be responsible for the binding to HDV RNA. To investigate HDAg sequences involved in HDV RNA binding, we synthesized five peptides, 15 to 29 residues in length, and tested their ability to bind HDV RNA using a simple non-radioactive ELISA with digoxigenin-labelled HDV genomic or antigenomic RNA probes. The specificity of interactions was demonstrated by comparison with control peptides and non-HDV RNA probes, and with an inhibition assay using recombinant HDAg. The HDAg-binding domain found within the middle region (79 to 163) of HDAg was more finely mapped: it is located between residues 79 and 107. In addition, another domain (residues 2 to 27) of HDAg was also found to bind specifically to HDV RNA. These two peptides share sequence similarities at residues 2 to 10 and 97 to 107 with other RNA-binding domains. PMID- 8245866 TI - Trans-activation of the adenovirus E2 promoter by human papillomavirus type 16 E7 is mediated by retinoblastoma-dependent and -independent pathways. AB - In common with the adenovirus E1A and simian virus 40 large T oncoproteins, the E7 protein of human papillomavirus (HPV) type 16 interacts with the retinoblastoma (Rb) tumour suppressor protein (pRb). The functional importance of this interaction for HPV-16 E7 protein was investigated by analysis of the transactivating function of E7 at the adenovirus E2 promoter in a set of breast tumour cell lines. Trans-activation by HPV-16 E7 in two pRb-deficient cell lines demonstrated that pRb is not essential for E7-mediated trans-activation, but reconstitution of Rb expression indicated the existence of an Rb-mediated pathway of E7 trans-activation. This pathway results from suppression by E7 of a trans repressing function encoded by the Rb gene. The E7 protein is shown to be capable of interacting in vivo with the Rb-related protein p107. Furthermore, analysis of a fusion construct between the amino terminus of Rb and the carboxy terminus of p107 suggests that, in common with pRb, the p107 protein trans-represses the adenovirus E2 early promoter. Therefore it is proposed that the pRb-independent pathway of E7 trans-activation is a consequence of the suppression of trans repression by p107. PMID- 8245867 TI - Characterization of an in vivo reactivation model of herpes simplex virus from mice trigeminal ganglia. AB - Herpes simplex virus type 1 (HSV-1) is transcriptionally active during latent infection in human peripheral sensory ganglia. Viral gene expression includes the latency-associated transcripts (LATs) which have been linked to the ability of the virus to resume replication and reactivate. However, the molecular basis of reactivation and the mechanisms of action of these transcripts are unknown. In order to study these parameters, an in vivo reactivation model is needed. We investigated use of the mouse as the experimental animal, modifying the route of infection, the viral strain and the reactivation protocol. Following administration of human immunoglobulin 1 day prior to corneal infection, no infectious virus was detected in trigeminal ganglia (TG). However, latency was established in all infected animals as indicated by explant reactivation of TG, and in vivo reactivation was achieved in 30 to 40% of them. DNA quantification revealed that TG of immunized mice contained more HSV-1 DNA than did those of non immunized mice. By in situ hybridization twice as many neuronal cells in TG of immunized mice were positive for LATs, compared with infected but non-immunized, mice. These findings suggest that suppression of primary infection facilitates reactivation by increasing HSV-1 copy number in latently infected nervous tissue. PMID- 8245868 TI - The herpes simplex virus type 1 strain 17 open reading frame RL1 encodes a polypeptide of apparent M(r) 37K equivalent to ICP34.5 of herpes simplex virus type 1 strain F. AB - The region between the 'a' sequence and the 5' end of the IE1 gene within the long repeat sequence of the herpes simplex virus (HSV) genome plays an important role in the neurovirulence of both HSV-1 strain F and HSV-1 strain 17. However, there has been controversy over the protein-coding potential of this region. Although an open reading frame (ORF) was predicted in HSV-1(F) and shown to encode a polypeptide called ICP34.5, only recently has a corresponding ORF, designated RL1, been recognized in HSV-1(17). To determine whether the HSV-1(17) ORF is expressed, we raised antipeptide sera against predicted amino acid sequences from RL1; one serum specifically recognized a 37K protein in HSV-1(17) infected cell extracts. Compared with the corresponding HSV-1(F) polypeptide the HSV-1(17) protein has a lower apparent M(r), shows similar kinetics of accumulation and intracellular localization but may accumulate to lower levels than the HSV-1(F) protein. The non-neurovirulent HSV-1(17) deletion variant 1716 fails to synthesize detectable levels of ICP34.5. Thus we have established that HSV-1(17), like HSV-1(F), expresses ICP34.5, a protein important for HSV neurovirulence. PMID- 8245869 TI - Murine cytotoxic T cell response specific for human cytomegalovirus glycoprotein B (gB) induced by adenovirus and vaccinia virus recombinants expressing gB. AB - A murine model of the cytotoxic T lymphocyte (CTL) response to glycoprotein B (gB) of human cytomegalovirus (HCMV) was developed based on the use of adenovirus (Ad) and vaccinia virus (Vac) recombinants expressing gB. Mice of different major histocompatibility haplotypes [CBA (H-2k), BALB/k (H-2k) and BALB/c (H-2d)] infected with the Ad-gB recombinant developed an Ad-specific CTL response. However, only the H-2k mice developed a significant HCMV gB-specific CTL response, as indicated by the major histocompatibility complex class I-restricted lysis of Vac strain Copenhagen (VacC)-gB recombinant-infected target cells by H 2k mouse immune spleen cells. The VacC-gB recombinant elicited only a weak gB specific CTL response in these mice, indicating that the observed gB-specific CTL response in mice is dependent on the expression vector used for immunization. The gB-specific cytotoxicity observed in H-2k mice was mediated by the CD8 lymphocyte subset. PMID- 8245870 TI - Comparison of 10 influenza A (H1N1 and H3N2) haemagglutinin sequences obtained directly from clinical specimens to those of MDCK cell- and egg-grown viruses. AB - PCR was used to amplify and sequence the complete HA1 region of the haemagglutinin (HA)-encoding genes of 10 clinical isolates of influenza virus of the H1N1 or H3N2 subtypes. These sequences were compared to those obtained from viruses isolated from the same specimens after passage in eggs and MDCK cells. Amino acid substitutions in the egg-derived HA sequences were found in nine out of the 10 specimens analysed, whereas seven out of eight of the MDCK-derived HA sequences were identical to those in the corresponding original specimens. Changes in the H1 HA occurred at residues 77a, 196 (also found in the corresponding HA from the MDCK isolate), 225, 226 and 227; changes in the H3 HA occurred at residues 137, 156, 186, 248 and 276. In addition, we have shown that an amino acid change at residue 145 in the HA of the H3 subtype that was previously demonstrated to be egg-selected is now present in circulating strains. PMID- 8245871 TI - Complete sequence conservation of the human T cell leukaemia virus type 1 tax gene within a family cluster showing different pathologies. AB - We have amplified, through PCR, the full-length tax gene of human T cell leukaemia virus type 1 (HTLV-1) derived from proviral DNA of peripheral blood lymphocytes of five first degree relatives of Afro-Caribbean origin. One patient (the father) had adult T cell leukaemia (ATL), one (the mother) tropical spastic paraparesis (TSP), and three (children) were healthy asymptomatic carriers. All five family members had identical tax nucleotide sequences as determined by direct sequencing of PCR products. This sequence was compared with tax gene sequences of an unrelated TSP patient of Afro-Caribbean origin, and of C8166 cells, and found to have one and seven nucleotide differences, respectively. At the amino acid level these three sequences differed from the HTLV-1 prototype Japanese strain (ATK-1). All sequence changes were clustered towards the 3' end of the gene. These data demonstrate the complete conservation of an HTLV-1 gene following, presumably, horizontal and vertical transmission of the virus. Clones of this gene showed more sequence variation within the TSP patient than the ATL patient, mostly consisting of point mutations; there was no conservation of mutations between the two individuals. These mutations occurred only in individual clones of the ATL patient whereas those of the TSP patient were found to be repeated in different clones. A tax-specific cytotoxic T lymphocyte response was observed in two asymptomatic carriers with low antibody titres, whereas none was detected in an individual with a high antibody level. No tax specific sequence was identified which may have contributed to the apparently high degree of transmission from mother to children (three of five children tested) nor account for the differences between disease symptoms in the parents. PMID- 8245872 TI - Antiviral therapy: the time has come. PMID- 8245873 TI - A retrospective and an observational study with acyclovir. AB - Retrospective analysis: This open controlled non-randomized study was carried out to investigate the influence of intravenous acyclovir (ACV) on the incidence of post-herpetic neuralgia (PHN). Twelve women and 11 men (mean age 52 years, range 19-89) received ACV 5 mg/kg every 8 hours) for 10 days (I). Twenty-seven untreated patients (mean age 62 years, range 20-89) were taken as a control group (II). Six to 24 months after the onset of herpes zoster (shingles) the patients were reexamined. The analysis revealed a significantly lower incidence of both general pain and severe pain (P < 0.05, chi 2 = 5.55 and 4.39) for (I) compared to (II). For 21 patients who were treated for a period of 10 days, the significance level was 1% (chi 2 = 7.82 and 8.62). Observational study: Fifteen thousand eight hundred and thirty-one non-hospitalized patients with shingles (mean age 55.2 years) received oral ACV (800 mg five times daily) for 7 days. At the onset of therapy, 15,420 patients (97.6%) reported pain (severe 42.6%, moderate 43.1%, mild 14.3%). The pain during treatment was documented by the patients (n = 5,728) in a diary and transferred to a scoring system (0 = none, 1 = mild, 2 = moderate, 3 = severe). From day 1 to day 7 there was a decrease in the pain score level from 2.3 to 0.9. Three months after the onset of herpes zoster, 2,519 of 14,858 patients (16.95%) reported pain; 311 patients (2.1%) complained of continuous pain, typical for PHN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245874 TI - Current management of cytomegalovirus disease. AB - Cytomegalovirus (CMV) is an important pathogen for the fetus, for allograft recipients, and for acquired immunodeficiency syndrome (AIDS) patients; the clinical features in each of these groups of patients are described. CMV may also act as a cofactor to accelerate the rate at which the human immunodeficiency virus (HIV) causes AIDS. Active CMV infection in immunocompromised patients is best managed by routine laboratory screening of patients at risk using assays that provide prognostic information. Antiviral therapy can then be considered under one of four headings: For prophylaxis, the drug is given to all patients from a particular time point, e.g., time of transplantation. Successful trials have been reported for interferon (renal transplant), acyclovir (renal transplant and bone marrow transplant), and ganciclovir (heart transplant and bone marrow transplant). For suppression, the drug is given once CMV excretion has been detected in peripheral sites of particular patients. A successful trial of ganciclovir in bone marrow transplant patients has been reported. For pre-emptive therapy, the drug is given once CMV has been detected systemically. A successful trial of ganciclovir in bone marrow transplant patients has been reported. Regarding treatment, a controlled comparison of ganciclovir and foscarnet for CMV retinitis in AIDS patients has shown that both drugs are equally effective but that foscarnet has a survival benefit. Open studies suggest that the combination of immunoglobulin and ganciclovir is of benefit for treatment of established CMV pneumonitis. In open studies other treatments have been tried, with no clinical success, despite control of viral replication in some cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245875 TI - Acyclovir influence on graft versus host disease. AB - Allogeneic bone marrow transplantation has been used for over two decades as a therapy to treat patients with malignant disease [Thomas et al., 1977; Geller et al., 1989; Clift et al., 1987; Gratwohl et al., 1990; Goldman et al., 1986; Thomas et al., 1986]. High doses of chemotherapy are administered either alone or in combination with total body irradiation in an attempt to eradicate malignant cells. The treatment may be lethal to normal bone marrow function, but this toxicity is overcome by providing bone marrow from an external source. In allogeneic bone marrow transplantation, bone marrow is obtained from an HLA identical family member or unrelated donor. In recent years the use of less well matched donors has increased, thus expanding the use of this strategy to a larger patient population. The success rate of allogeneic bone marrow transplantation has been greatest in the treatment of haematopoietic malignancies. Patients with acute or chronic leukaemia have a 30-80 percent likelihood of being free of disease at 5 years following transplantation. The success rate depends on the stage of disease at the time of transplantation. Certain nonmalignant diseases have also been treated successfully with allogeneic bone marrow transplantation. These include severe aplastic anaemia, inborn errors of metabolism, and other genetically determined diseases [Storb et al., 1986a, 1991; Lucarelli et al., 1990; Kirkpatrick et al., 1991]. With the availability of effective antiviral therapy, treatment and prophylaxis are available for HSV, CMV, and VZV. Acyclovir has been shown to be effective in treating established infections with HSV and VZV, and in the prophylaxis against HSV, severe CMV infections, and VZV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245876 TI - Acyclovir in preventing cytomegalovirus infection in kidney transplant recipients: a case-controlled study. AB - High dose oral acyclovir has been reported to be effective in preventing both cytomegalovirus (CMV) infection and disease in renal transplant recipients. We conducted a case-controlled study in which 42 cadaveric kidney transplant recipients were prophylactically treated with high dose oral acyclovir for 3 months and compared to historical controls matched for donor/recipient CMV serological status, age, sex, and immunosuppressive therapy. Before transplantation, study group patients received acyclovir intravenously (500 mg/m2 over 1 hour) which was subsequently given orally (basal dose--800 mg four times daily) from day 2 post-transplantation according to renal function. All patients received 14-day induction immunosuppressive therapy with either a polyclonal or a monoclonal antibody together with low dose steroids and azathioprine, cyclosporin being introduced at day 10 post-transplantation. Blood viral cultures as well as CMV antibody titers were performed in study group and control patients in the same laboratory, before transplantation, weekly until 3 months and then monthly until 6 months. CMV infection was defined as a positive blood or bronchoalveolar lavage viral culture or presence of CMV IgM or CMV IgG in a previously seronegative patient. Diagnosis of CMV disease also required the presence of at least one concomitant febrile illness, with or without other clinical symptoms, not attributable to another pathogen. All patients were followed for 3 months. Incidence of both CMV infection and disease was compared in the two groups using the log-rank test. With regard to CMV infection, we found significantly less CMV infection in CMV seropositive patients (regardless of donor CMV serological status) in the study group compared to historical controls (P = 0.005).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245877 TI - Solid organ transplantation: results and implications of acyclovir use in liver transplants. AB - CMV infection is a major cause of morbidity and mortality following liver transplantation (LT). A prospective study of 218 LT recipients showed that 55% of patients developed CMV infection during the 1st year post-transplantation. Symptomatic CMV infection developed in 25% of all patients, being a major cause of death (21% of all deaths). Of 62 episodes of documented organ invasion, liver was the major site (38 episodes), followed by lung (20), gastrointestinal (4), and retina (4). The main patient group at risk (according to CMV serology of the recipient [(R)/donor(D)]) was the R-/D+: 77% of patients developed CMV infection, all of them with symptoms. The lowest group at risk was the R-/D-: 13% of patients developed CMV infection, half of whom developed symptoms. Time-dependent multivariate statistical analysis of risk factors indicated that the R-/D+ group was the main risk factor for CMV infection (P < .02) and symptomatic infection (P < .0001). To decrease the incidence and severity of CMV infection following LT, a randomized study is ongoing to evaluate the efficacy of ganciclovir (5 mg/kg/IV/q 12 hours for the first 14 days post-LT) followed by acyclovir (800 mg/po/qid for 14 weeks) GCV + ACV (group I), versus acyclovir (same dose for 16 weeks, starting immediately post-LT) ACV (group II). These treatment groups are compared to matched historical controls (C). Preliminary analysis of 83 LT recipients indicates that in group I the median date for the first evidence of CMV infection is delayed (82 days) as compared to group II and C (41 and 33 days, respectively) (P = .004).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245878 TI - Cytomegalovirus infection in acquired immune-deficiency syndrome. AB - Cytomegalovirus (CMV) infection is very common in acquired immune deficiency syndrome (AIDS) with approximately 40% of patients presenting CMV visceral involvement at advanced stage disease. The most common localizations are retinitis and gastrointestinal involvement; central nervous system disorders are likely to be underestimated. Diagnosis of CMV disease should be assessed, for all localizations except retinitis, on the association of clinical symptoms with presence of CMV inclusions in biopsy specimens. Two drugs, ganciclovir and foscarnet, are currently licensed for treatment. Both are administered intravenously. Since progression of CMV disease is very common particularly in retinitis, maintenance therapy should be given throughout remaining life. Unfortunately, maintenance therapy does not entirely prevent the risk of relapses. Drug toxicity is mainly bone marrow suppression with ganciclovir and renal insufficiency with foscarnet. Since quality of life is severely impaired by long-term daily intravenous treatment, orally active drugs are urgently needed both for treatment and primary prophylaxis of CMV infection. PMID- 8245879 TI - Neonatal herpes simplex virus infections. AB - Neonatal herpes simplex virus infections are a common problem in the United States, occurring at an incidence of one in approximately 3,500 deliveries. In the absence of antiviral therapy significant morbidity and mortality is attendant with disease. Disease manifestations in the newborn include multiorgan involvement (disseminated disease), encephalitis, and/or infection limited to the skin, eye, or mouth. These three broad classifications provide distinctions in severity of disease for evaluation of outcome following antiviral therapy. The availability of antiviral therapy for life-threatening disease, particularly that which is disseminated or involves the brain, has been of particular benefit for children with neonatal herpes. Both acyclovir and vidarabine have proven effective in the management of neonatal herpes simplex virus infection. With current therapeutic modalities, mortality from disease localized to the skin, eye, and mouth is virtually non-existent, yet a few children (approximately 5%) are still at risk for a long-term neurologic sequelae. For babies with encephalitis, the mortality has been reduced to approximately 15% and nearly 50% of survivors develop normally 3 years after treatment. Outcome with disseminated infection is of less value as mortality remains high (50%), but the number of survivors who develop normally is approximately 85%. The introduction of new antivirals with enhanced lipophilicity and, potentially, greater activity in the central nervous system may further improve outcome from this devastating disease. PMID- 8245880 TI - The acyclovir legacy: its contribution to antiviral drug discovery. AB - The discovery of acyclovir marked the beginning of an exciting era in antiviral research. Early studies on the novel mode of action explained the selectivity of the compound and the remarkably narrow spectrum of activity against a subset of the herpesviruses. Throughout the past decade many clinical trials have demonstrated the efficacy and safety of this drug. Furthermore, the development of resistance does not appear to be a significant issue in normal individuals. Acyclovir provided the stimulus for further work in the antiherpes area, and this has led to the recent discovery of an oral prodrug (256U87), which delivers higher levels of acyclovir by the oral route, and to the discovery of 882C87, a highly selective inhibitor of varicella zoster virus. The novel mode of action of acyclovir involves an extremely selective phosphorylation step carried out by the herpesvirus thymidine kinase. It has recently been shown with another nucleoside analogue, ganciclovir, active against human cytomegalovirus (HMCV), that activation can be carried out by other unrelated kinases (in this case the UL97 gene product). Studies of this type may lead to the development of further novel inhibitors. PMID- 8245881 TI - Review of research leading to new anti-herpesvirus agents in clinical development: valaciclovir hydrochloride (256U, the L-valyl ester of acyclovir) and 882C, a specific agent for varicella zoster virus. AB - Research leading to the new anti-herpesvirus compounds discussed here has come from three approaches. The first approach was directed towards improving the bioavailability of acyclovir by examining the potential of a variety of prodrugs, leading to the new compound valaciclovir hydrochloride. The second approach was to examine a large number of 5-substituted pyrimidines for activity against those viruses which were not as potently inhibited by acyclovir as are herpes simplex viruses, i.e., varicella zoster virus (VZV) and human cytomegalovirus (HCMV). This research led to the new chemical entity 882C for VZV. A third approach has been to examine drug combinations with acyclovir. This research led to the compound 348U, an inhibitor of herpes simplex virus ribonucleotide reductase which acts synergistically in combination with acyclovir. This manuscript will focus on the first two approaches leading to new compounds valaciclovir hydrochloride and 882C since Dr. Safrin details such background for 348U/acyclovir. Attempts to improve the bioavailability of acyclovir began a decade ago. Early prodrugs were compounds with alterations in the 6-substituent of the purine ring of acyclovir. The 6-amino congener required the cellular enzyme adenosine deaminase for conversion to acyclovir and the 6-deoxycongener was dependent on cellular xanthine oxidase for conversion. Neither of these prodrugs had a chronic toxicity profile in laboratory animals as good as acyclovir. Efforts were directed towards simpler esters and 18 amino acid esters were made. The pharmacokinetic profile of each prodrug was determined in rats by measuring the recovery of acyclovir in urine after oral dosing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245882 TI - Potential for combined therapy with 348U87, a ribonucleotide reductase inhibitor, and acyclovir as treatment for acyclovir-resistant herpes simplex virus infection. AB - Inhibitors of the ribonucleotide reductase of herpes simplex viruses (HSV) potentiate the activity of acyclovir in vitro and in animal studies. In addition, the combination of the ribonucleotide reductase inhibitor 348U87 and acyclovir has synergistic therapeutic effects against infections in mice due to thymidine kinase-deficient, thymidine kinase-altered, and DNA polymerase mutants of HSV. We performed a pilot study of topical combination therapy with 348U87 (3%) and acyclovir (5%) cream for acyclovir-resistant, anogenital HSV infections in ten human immunodeficiency virus (HIV)-infected patients. Our results, with lack of complete reepitheliazation of lesions in all patients and poor virologic response, suggest that this therapy is unlikely to be useful for this indication. PMID- 8245883 TI - Valaciclovir (BW256U87): the L-valyl ester of acyclovir. AB - Valaciclovir (BW256U87) is an L-valyl ester of acyclovir, which is extensively and almost completely converted to acyclovir. In healthy human volunteers, single valaciclovir doses of 100-1000 mg resulted in dose-proportional increases in acyclovir area under the curve (AUC). The 1,000 mg dose produced an acyclovir peak plasma concentration (Cmax) of 5-6 micrograms/ml, AUC6 of 19 hr. micrograms/ml, time to maximum plasma concentration (Tmax) of 1-2 hr, and half life (T1/2) of 2.8 hr. Plasma valaciclovir peak levels were < 0.3 micrograms/ml, and the prodrug was undetectable after 3 hr. Multiple valaciclovir doses of 250 2,000 mg given four times daily for 10 days resulted in dose-proportional increases in acyclovir Cmax. There were less than proportional increases in the AUCs. No serious or unexpected adverse events or laboratory abnormalities were reported. In volunteers with advanced human immunodeficiency virus (HIV) disease (absolute CD4 lymphocyte count < 150 cells/microliters), acyclovir and valaciclovir pharmacokinetic results were nearly identical to those in healthy volunteers. At the 2 g dose administered four times daily, steady-state acyclovir Cmax = 8.4 micrograms/ml, Tmax = 2.0 hr, AUC6 = 30.5 hr. micrograms/ml, and T1/2 = 3.3 hr. Nausea, vomiting, diarrhoea, and abdominal pain were commonly reported; however, only one adverse event (diarrhoea) was causally linked to valaciclovir exposure. There were no renal or neurologic adverse events. Valaciclovir is well absorbed and is rapidly converted to acyclovir, resulting in three- to fourfold higher acyclovir levels than can be achieved with oral acyclovir, even in patients with advanced HIV disease. The safety profile is generally favourable, with no evidence of nephrotoxicity or neurotoxicity. PMID- 8245884 TI - Preliminary pharmacokinetics and safety of 882C87 in patients with herpes zoster. AB - 882C87 [1-(beta-D-arabinofuranosyl)-5-propynyluracil] is a nucleoside analogue with potent and specific antiviral activity against varicella-zoster virus (VZV). The IC50 of 882C87 against VZV ranges from 0.6 to 3.8 microM. Potentially therapeutic plasma concentrations are readily achieved in humans; the pharmacokinetics have been previously evaluated in healthy young and elderly (> 65 years) volunteers following single oral doses of 50-400 mg. Thirty immunocompetent patients with localised herpes zoster were treated with 882C87. Groups of patients received 50 mg, 100 mg, or 200 mg tablets of 882C87 every 12 hours for 7 or 7.5 days (14 or 15 doses). Six patients in each group were over 60 years of age. Blood samples for determination of 882C87 concentrations were taken at entry, steady state, and during the elimination phase following the last dose. After the final doses of the 50 mg 100 mg, and 200 mg dosage regimens, the Cmax of 882C87 in patients over 60 years old was 7.7 +/- 3.1 microM, 12.6 +/- 3.5 microM, and 24.8 +/- 14.0 microM, respectively, and the AUCs 0-12 were 78.4 +/- 31.8 microM.hr, 137.5 +/- 22.8 microM.hr, and 272.5 +/- 170.5 microM.hr, respectively. Preliminary estimates of the elimination half-life ranged from 15.1 to 20.0 hr. These preliminary pharmacokinetic data confirmed good dose proportionality for AUC and Cmax with values between those predicted from single dose data in the young and those in elderly volunteers. The plasma concentration profiles at these doses were in excess of IC50 values and support the use of once or twice-daily regimens in future studies of 882C87 in herpes zoster. PMID- 8245885 TI - Impact of herpesvirus infections in the future. AB - Numerous diverse, often opposing, trends will determine the frequency (and severity) of herpesvirus infections in the future. A major factor is the growing population of iatrogenically immunocompromised patients, accruing especially from large increases in organ and autologous bone marrow transplantation. Of even greater importance is the immense number of human immunodeficiency virus (HIV) infected patients for whom herpesviruses may: (1) have been a cofactor in acquisition of HIV infection; (2) contribute to in vivo activation of HIV; (3) cause severe infections. Another factor influencing severe herpesvirus infections is the aging of populations in developed countries; this will be associated with a greater prevalence, and more morbid manifestations, of herpes zoster. Sociologic changes will also be important. The consequences of these, such as attempts to influence sexual practices, will be difficult to predict. Other changes, such as more frequent use of early child care facilities, will predictably lead to early acquisition of most herpesviruses, thereby decreasing the incidence of severe disease in adulthood. Factors that will reduce the incidence and/or severity of herpesvirus infections include vaccines (varicella, herpes simplex, cytomegalovirus); prophylactic strategies for immunocompromised hosts (passive immunization, antiviral drugs, blood product selection); more rapid and sensitive diagnostic methods; and suppressive or early antiviral therapy for common infections, such as genital herpes simplex and varicella.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245886 TI - Future management of herpesvirus infections. AB - The clinical investigations conducted to date with herpesviruses have provided a good grounding into the general principles of herpesvirus replication and pathogenesis. Multiple opportunities for intervention include prevention of initial infection, prevention of reactivation, suppression of reactivated virus, and treatment of established disease. Studies have shown that each of these approaches can be effective against one or more herpesviruses. What is needed now are studies combining sequential interventions so that patients who fail, for example, prophylaxis with one drug can be entered directly into trials of suppression and then into pre-emptive therapy, each with different antiviral agents. In this way, it is to be hoped that increasing proportions of cohorts of susceptible patients will suffer less from herpesvirus infections. Clearly, the design of such clinical trials needs to be constantly refined as methods of diagnosis and understanding of pathogenic mechanisms improve and as new agents are discovered. PMID- 8245887 TI - Acyclovir: discovery, mechanism of action, and selectivity. AB - The reasons for acyclovir's activity and selectivity in cells infected with HSV or VZV may be summarized as follows: 1. Activation by a HSV- or VZV-specified TK. 2. Greater sensitivity of viral DNA polymerase than of the cellular polymerases to ACV-TP. 3. Inactivation of the viral DNA polymerase, but not the cellular polymerases, by ACV-TP. 4. Chain termination of viral DNA by incorporation of ACV MP. For the Epstein-Barr virus, which is also sensitive to acyclovir, there is no selective activation in infected cells [Colby et al., 1981], but the viral polymerase can be inhibited by very low levels of ACV-TP [Datta et al., 1980]. For HCMV, the activation of acyclovir is very poor but the viral polymerase is also more sensitive to ACV-TP than the cellular polymerases. One of the important contributions of acyclovir was the demonstration for the first time that a compound could prevent the DNA replication of a DNA virus at concentrations far below those that affect cellular DNA synthesis. As we all know, in the past 15 years there has been a complete rejuvenation of antiviral chemotherapy. I think it is very fortunate that we changed our outlook on the possibility of making potent and selective antiviral agents in time so that, when the AIDS epidemic came along, we did not feel completely at a loss on ways to attack viral disease. PMID- 8245888 TI - Natural history, management and complications of herpes labialis. AB - Infection with herpes simplex virus (HSV) is a common worldwide problem. Primary infection with HSV-1 rarely causes significant problems although widespread involvement in atopic eczema can be life-threatening as may associated encephalitis. Keratoconjunctivitis, pharyngitis and hepatitis can also complicate primary infection. Twenty to 40% of the population at some stage have recurrent orolabial infections with HSV although in only 1% of these cases is this recurrence severe. Recurrent erythema multiforme appears to be associated with HSV-65% of patients are thought to have preceding herpes labialis. Many primary and recurrent infections with HSV-1 require little more than topical antiseptic therapy to control secondary infection. Systemic acyclovir, however, is indicated in various situations including complicated primary infection, infection in neonates, eczema herpeticum, HSV infections in the immunocompromised, and recurrent erythema multiforme. In the latter, prophylactic treatment with 6 months acyclovir appears to be effective. PMID- 8245889 TI - Prophylactic chemotherapy with acyclovir for recurrent herpes simplex labialis. AB - The medical literature was reviewed and 11 clinical trials of prophylactic topical or peroral acyclovir for the suppression of recurrent herpes simplex labialis were identified. The results of these trials showed that prophylactic topical acyclovir was mostly ineffective, but that prophylactic peroral acyclovir, in doses ranging from 400 to 1,000 mg/day, reduced the frequency of herpes labialis during treatment by 50-78%. The reduction in the frequency of episodes of herpes labialis with acyclovir prophylaxis is less than the suppressive effect that has been reported for herpes genitalis (50-78% vs. 80 90%). In trials of prophylactic acyclovir for herpes labialis induced by experimental ultraviolet radiation, 26% of induced lesions developed within 48 hours of radiation exposure ("immediate" lesions) and, in contrast to "delayed" lesions that developed 2-7 days post-irradiation, were not suppressed by the antiviral compound. It is proposed that these treatment-unresponsive immediate lesions have an atypical pathogenesis, possibly involving latency of herpes simplex virus in the labial epithelium, and that these may be responsible for the apparent difference between herpes labialis and genitalis in the degree of benefit from prophylactic acyclovir therapy. PMID- 8245890 TI - Genital herpes: natural history and treatment of acute episodes. AB - Genital herpes is a common, distressing infection whose incidence has been underestimated. Recent serological surveys, employing type-specific antibody assays, show a rising prevalence of previous herpes simplex virus (HSV) type 2 infections in post-adolescent populations in developed countries; many of these infections have been asymptomatic. In some geographical locations, HSV-1 infections are a common cause of first episodes. They may occur in stable, monogamous relationships and are less likely to recur than genital infections caused by HSV-2. The clinical features of first episode genital herpes show marked individual variation in severity; they tend to be more severe in women than in men. Local and distant complications are common in immunocompetent individuals and may be life-threatening in those who are immunocompromised. The psychological and social consequences which result from the life-long infection and its risks of transmission to new sexual partners often prove more disabling to affected individuals than do the milder physical symptoms associated with recurrent episodes. Counselling, support, and patient education are essential components of management. Acyclovir is clearly established as the first choice therapy in both first and recurrent episodes of genital herpes. This drug has potent antiviral effect and provides significant clinical benefit in first episodes. Systemic therapy for initial episodes does not prevent either the establishment of latency or the development of future recurrences even when used in high or prolonged dosage. Episodic treatment of recurrences, with either oral or topical acyclovir, requires early patient initiation of therapy to provide significant clinical benefit. PMID- 8245891 TI - Long-term clinical and psychological management of genital herpes. AB - Many patients with genital herpes will have recurrences, and for some the recurrences may be frequent and severe and accompanied by profound psychosexual morbidity. Some patients can successfully be managed with intermittent courses of oral or topical acyclovir to be used with each recurrence. However this treatment is of limited efficacy, and longterm acyclovir suppression may be useful. This form of treatment is highly successful for patients with frequent recurrences most of whom will have no episodes during treatment. The optimum dosage for commencing acyclovir suppression is 200 mg four times daily, and the dose may subsequently be reduced. Some patients can successfully be managed on 400 mg twice daily. Treatment should be stopped after 1-2 years as there is some evidence of a decrease in the frequency of recurrences. Selecting patients for suppression should be based on the frequency, severity, and duration of recurrences as well as any associated emotional problems. Some patients may recur on suppression, usually due to inadequate dosage. Many patients with first episode genital herpes experience emotional and psychological problems. However these problems only continue if patients have recurrences and continued psychological support is essential. Long-term acyclovir suppression improves psychological well-being but is not a substitute for information, counselling, and expert psychosexual support. PMID- 8245892 TI - Continuous five-year treatment of patients with frequently recurring genital herpes simplex virus infection with acyclovir. AB - This study presents data relative to the efficacy and safety following the continuous use of oral acyclovir in the treatment of genital herpes over a 5-year period. In this study, 1,146 patients (53% males; 47% females) were originally enrolled. These included patients with a history of frequently recurring genital herpes (mean > 12 episodes per year). During the first year, patients were randomized between those receiving 400 mg of acyclovir twice daily and an equal number receiving placebo. Additionally, acyclovir was utilized for episodic treatment (ES) in both groups. Thereafter, patients received open-label acyclovir suppressive therapy for the remainder of the study period. Complete data are available on 389 patients who completed the fifth year of therapy. All the participants who completed the fifty year of the study had completed either 4 or 5 years of daily suppressive acyclovir therapy. During the first year, a significant decrease in the frequency of recurrences in patients receiving continuous acyclovir (SS) was noted as compared to the placebo group (1.7 vs. 12.5 recurrences; P < 0.0001). From year one to the end of year three, a progressive decrease in the frequency of recurrences was noted in both groups. Yet, those patients who had received SS for the full 3 years had significantly fewer recurrences than those who had received ES in the first year (P = 0.05). During years four and five, the decrease in frequency of recurrences between the ES and SS groups was not significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245893 TI - Assessing the economic benefits of antiherpes therapy. AB - Increasing pressures on health-care budgets mean that health-care treatments and programmes need not only to demonstrate that they are efficacious, but also that they deliver good value for money. In the context of pharmaceuticals it is likely that both in Australia and Ontario (Canada) evidence of cost effectiveness will become a mandatory requirement prior to reimbursement (government subsidy) of drugs. However, in most jurisdictions economic data will be one of a number of factors taken into account by decision makers, be they policy makers, members of formulary committees, or individual prescribers. Methods of economic evaluation have been developed for the assessment of healthcare treatments, but these have not been extensively applied to antiherpes therapy. However, this paper, based on a literature review of the clinical trials of acyclovir, shows that there are many indicators of potential economic benefit, depending on the clinical indication, in length or quality of life, savings in other health-care resources, or wider benefits (e.g., reductions in lost work time). If appropriate economic data are to be gathered in future for new antiherpes therapy, a more systematic approach is required, conducting economic evaluation alongside, or as an integral part of, clinical trials. This raises important logistical and methodological challenges. It is important that researchers acknowledge these challenges early so that they can be adequately addressed. PMID- 8245894 TI - Sensitivity monitoring of clinical isolates of herpes simplex virus to acyclovir. AB - Zovirax (acyclovir, ACV) is now widely accepted as a safe and effective treatment for the management of herpes simplex virus (HSV) infections in normal and immunocompromised patients. However, a common concern with regard to the widespread use of any antimicrobial agent is resistance. The virus specific mechanism of action of ACV involves two virus encoded enzymes, thymidine kinase (TK) and DNA polymerase. Any alteration in the genes coding for these two enzymes would therefore be expected to confer resistance. The findings from two extensive resistance monitoring programs have shown that in immunocompetent patients receiving ACV for the management of acute HSV disease, the incidence of resistance is extremely rare. The situation in the immunocompromised is different. In this patient group HSV disease is severe and protracted often requiring prolonged therapy thus increasing the exposure of the virus to drug. As a result HSV isolates resistant to ACV have occasionally been recovered. Biochemical and genetic analysis of the resistant clinical isolates has shown that resistance in the most part is due to an inability of the virus to produce TK which mirrors the findings with cell culture derived resistant virus. Laboratory studies would indicate that TK-deficient virus would have little clinical impact. Significantly, resistance has rarely been attributed to alterations in the substrate specificity of TK or DNA polymerase. The biological significance of these mutants is unclear but to date there has been no evidence of transmission of resistant virus. PMID- 8245895 TI - Safety of acyclovir: a summary of the first 10 years experience. AB - The era of pharmacoepidemiology has introduced profound changes in the ability of the sector to monitor the post-marketing safety of new products. We report on the population-based assessment by Wellcome of the safety of Zovirax reflected in a program of unprecedented scope, diversity, and, most important, utility. The program couples epidemiologic intelligence--analysis of adverse experience reports arising spontaneously from practice and reported directly to the manufacturer, regulators, or in the published literature--with more structured formal epidemiologic research approaches. Taken together, the absence of major medical problems emerging from the monitoring system for spontaneous reports from the medical practice experience among over 20 million persons treated worldwide over the past decade, complemented by the similar absence of "signals" from structured epidemiologic studies closely monitoring the experience of over 50,000 patients, constitutes a persuasive body of information concerning the general safety of this important therapeutic intervention. PMID- 8245896 TI - Herpes simplex virus infections during the decade since the licensure of acyclovir. AB - The development and use of acyclovir accelerated in the area of the natural history, epidemiology, and biology of herpes simplex viruses. The development of serologic assays that differentiate HSV-1 from HSV-2 demonstrated the worldwide distribution of genital herpes, the role genital ulcers play in facilitating HIV transmission, and the high frequency in which pregnant women acquire HSV infection. The high reactivation rate of HSV-2 infections, the often silent nature of genital herpes, and its widespread anatomic distribution in the genitourinary tract lead to frequent subclinical transmission. Identification and counselling of the asymptomatic carrier is necessary if we are to decrease the transmission of genital herpes. PMID- 8245897 TI - Current management of varicella zoster virus infections. AB - A series of randomized, placebo-controlled, double-blind clinical trials conducted from 1980 to the present provide the basis for appropriate management of varicella-zoster virus (VZV) infections. Placebo recipients in these studies have also provided valuable natural history data on the clinical course of VZV infections. The protocols in toto have shown acyclovir (ACV) to be safe and effective for treatment of nearly all forms of acute VZV infection. A number of issues still need to be addressed, including appropriate dosage, importance of early initiation of therapy, cost-benefit ratio, and viral resistance. Considering the data in aggregate, the author recommends ACV treatment for all acute VZV infections in immunocompromised hosts; for acute herpes zoster infections in all adults; and for varicella in otherwise healthy adults and adolescents, and children who contract it from a sibling. PMID- 8245898 TI - Varicella-zoster virus infection in immunocompromised patients. AB - The prophylactic effect of acyclovir (ACV) on varicella-zoster virus (VZV) infection in leukaemia patients who have undergone bone marrow transplantation (BMT) was reviewed. The benefits of the use of the laminar air flow (LAF) room in the prevention of nosocomial VZV infections in the haematological ward are also discussed. Since 1986 ACV has been administered to BMT patients to prevent herpes simplex virus (HSV) infections. Of 98 patients with leukaemia who underwent BMT, 73 received ACV (200 mg five times daily) and 25 were not given ACV. In the untreated group, 9 patients (36.0%) developed VZV infection by day 67 (median) and 3 patients died due to disseminated VZV infection. In the ACV-treated group, 18 patients (24.6%) developed VZV infection by day 150 (median) and there were no deaths. From July to December 1989, nine cases of VZV infections (eight patients and one nurse) were reported in the haematological ward of the hospital. All cases originated in the conventionally ventilated areas of the ward while no VZV infections were reported in the 14 patients who occupied the LAF rooms during the same period. PMID- 8245899 TI - Acyclovir therapy for varicella in otherwise healthy children and adolescents. AB - Acyclovir has been approved in the United States and elsewhere as antiviral therapy for otherwise healthy children and adolescents with varicella. This development arose from multicentre placebo-controlled trials of acyclovir in normal patients, 2-18 years of age, which showed that the drug accelerated cutaneous healing, and reduced fever and related constitutional symptoms without harmful side effects. Acyclovir did not, however, decrease transmission of chickenpox within the household, nor was there any demonstrable effect of antiviral therapy on varicella complications. In this article, the background and rationale for the multicentre studies of acyclovir in normal paediatric patients with chickenpox is reviewed. The evidence for and against its routine administration within 24 hours of the eruption of skin rash is also discussed. PMID- 8245900 TI - Treatment of varicella in the immunocompetent adult. AB - Varicella in the immunocompetent adult is an infrequent but potentially serious infection. Previous studies in immunocompetent hosts and normal adults have demonstrated the value of intravenous acyclovir in the treatment of varicella zoster virus infections. Oral acyclovir has also shown efficacy in both normal adults with zoster (shingles) and immunocompetent children with varicella. A recently completed double-blind placebo-controlled study of oral acyclovir in immunocompetent adults with uncomplicated varicella also demonstrated efficacy. Therapy within the first day reduced the time to 100% crusting of skin lesions from 7.4 to 5.6 days, and reduced the duration of fever by one-half day. Symptoms were also diminished. These benefits were observed only when therapy was initiated within 24 hours of the appearance of the rash. Adults with complicated varicella (usually symptomatic varicella pneumonia) should receive intravenous acyclovir. Several new agents for varicella-zoster therapy are being evaluated; brovavir is a new agent currently being compared to placebo in the treatment of adult varicella. PMID- 8245901 TI - Effect of oral acyclovir on pain resolution in herpes zoster: a reanalysis. AB - The most frequent complication of herpes zoster is postherpetic neuralgia, usually defined as chronic pain in the area of the exanthem that persists for at least a month after the skin lesions have healed. Several clinical studies of acyclovir showed a reduction in severity and duration of acute pain, but provided no definitive data for chronic pain. In order to determine if acyclovir therapy could reduce chronic pain, we reanalyzed data from the largest U.S. placebo controlled treatment trial of 187 immunocompetent persons with herpes zoster. By considering pain as a continuum, we found that the median duration of pain in acyclovir recipients was 20 days vs. 62 days for their placebo counterparts (P = 0.02). Thus, acyclovir has been shown to reduce chronic zoster-associated pain. We also noted that the absence of pain at the onset of cutaneous herpes zoster did not preclude its later development. PMID- 8245902 TI - Management of ophthalmic zoster. AB - The natural history of herpes zoster ophthalmicus and aspects of its treatment and prevention are presented. Intraocular complications occur in 50 percent of cases. Anterior uveitis and the various varieties of keratitis are commonest, affecting 92% and 52% of patients with ocular involvement, respectively. Sight threatening complications include neuropathic keratitis, perforation, secondary glaucoma, posterior scleritis/orbital apex syndrome, optic neuritis, and acute retinal necrosis. Twenty-eight percent of initially involved eyes develop long term ocular disease (6 months), with chronic uveitis, keratitis, and neuropathic ulceration being the commonest. Acute pain occurs in 93% of patients and is still present in 31% at 6 months. Of patients aged 60 and over pain persists in 30% for 6 months or longer, and this rises to 71% in those aged 80 and over. Current evidence favours the use of topical acyclovir alone for treatment of established ocular complications, with topical steroids being withheld in all but the most severe cases. Stellate ganglion block has proved useful in the treatment of established acute pain. Amitryptiline, and to a lesser extent sodium valproate, are useful in established chronic pain. Evidence of the efficacy of early oral acyclovir on ocular complications is conflicting, with two studies reporting significant improvement in differing disease parameters. A similar situation exists for pain, with published studies showing differing effects on pain at varying times after the onset of disease. The use of systemic steroids to prevent pain is not supported by currently available evidence, but its therapeutic relationship with acyclovir requires further evaluation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245903 TI - Motivational orientation and span of apprehension in children with mental retardation. AB - Children diagnosed as mildly mentally retarded were examined with respect to performance on Estes's (1965) span-of-apprehension task. Based on their scores on the Simplified Version of the Intellectual Achievement Responsibility scale, we divided subjects into a "learned-helpless" group and a "mastery-oriented" group. Motivational orientation had a significant effect on performance, with the mastery-oriented subjects demonstrating higher detection accuracies than the learned-helpless subjects. These results have implications regarding not only centrally mediated attentional functioning in children with mental retardation, but also interpretation of certain previous findings with the span-of apprehension task. PMID- 8245904 TI - Freud, Piaget, or neither? Beliefs in controlling others by wishful thinking and magical behavior in young children. AB - Tests showed that 4- to 6-year-old children believe that people can be influenced or "controlled" both by thinking (e.g., both wishful thinking and magical behavior) and by realistic means (e.g., positive reinforcement, example, and group pressure). Belief in control by thinking did not vary by the subjects' sex or age, influence type (wishing or magical behavior), or target response (behavior, emotion, or thought). Quantitative measures, however, suggested that magical behavior was seen by subjects as being more efficient than wishing, and that emotion was considered easier to influence than thinking. Beliefs in control by thinking were not related to a measure of fantasy-reality differentiation (realism). PMID- 8245905 TI - Predictive associations between Type A behavior of parents and their children: a 6-year follow-up. AB - This study examined over a period of 6 years whether or not Type A behavior in parents predicts Type A behavior in their children. The subjects comprised 1,116 originally randomly selected 12-, 15-, and 18-year-old adolescents and their parents. Type A behavior of the adolescents was self-rated, using two different methods: the Type A Behavior Questionnaire for the Finnish Multicenter Study (Keltikangas-Jarvinen & Raikkonen, 1990) and the Hunter-Wolf A-B Rating Scale (Wolf, Sklov, Wenzl, Hunter & Berenson, 1982). Parents already had rated their Type A behavior with the Framingham Type A Scale (Haynes, Levine, Scotch, Feinleib, & Kannel, 1978) 6 years earlier. The results indicated that children are subject to the influence of both parents. However, the study found parent child interactions to be dimension-specific and dependent on the sex of the parent. Therefore, the findings emphasize that in studying the parent-child Type A similarity, the multi-dimensionality of Type A behavior and the conjoint influence of parental behaviors should not be forgotten. PMID- 8245906 TI - Relationships between depression and attachment styles in parent- and institution reared Turkish children. AB - The present study, using a sample of Turkish children, aimed to test two attachment-theory predictions: (a) institution-reared children who were separated from their parents at an early age are less likely to have secure attachment schemata than children from two-parent families, and (b) depressive schemata are positively related to insecure attachment schemata and negatively related to secure attachment schemata. Following a systems-theory perspective, we also predicted that the relationship between depression and attachment styles would be stronger for institution-reared children than for children from two-parent families. Hazan and Shaver's (1987) attachment-style items and the Beck Depression Inventory were administered to 20 institution-reared and 20 parent reared boys by a female interviewer. The results supported the first two predictions of the study. PMID- 8245907 TI - Referential set presentation effects on complementation by 6-year-olds. AB - What difficulties do children encounter when responding to complementation instructions? The responses of 50 children to six complementation requests were examined. For half of the children, the objects in the referential set were pooled, and for the other half, they were physically separated by category (pencils were presented in a pencil box, buttons in a sewing kit, and balls of yarn in a knitting basket). Reinforcing the identifying status of the semantic property by separating the objects into categories led to an increase in the number of responses bearing on the subset designated in the instructions (e.g., in response to "Give me everything that is not a black button," the children handed over only the nonblack buttons). In contrast, older children gave responses based on the entire set. The meaning attributed by children to object properties and the effects of that attribution on processing mode merit further study. PMID- 8245908 TI - Laterality of cradling in relation to perception and expression of facial affect. AB - A dominant leftward cradling bias has been observed in women in non feeding interaction with infants. Reasons for this behavior have been sought in behavioral asymmetries, but none have sufficiently justified the presence of this leftward pattern. Recently, the cradling bias has been linked to affective processing, considered to be a specialized function of the nondominant (right) hemisphere (Manning & Chamberlain, 1990). This study investigates Manning and Chamberlain's (1990) suggestions that a relationship exists between leftward cradling and the interpretation and expression of affect. Eighty-six nulliparous women were tested for this relationship by correlating direction of cradling bias with visual field dominance for perception of facially expressed emotion and expression of affect. No significant relationship was found to directly support the hypotheses. PMID- 8245909 TI - Marital dissolution by sex of the petitioner: a test of the man-child affiliative bond. AB - The problems in disentangling the tabula rasa explanations, as opposed to the biocultural explanations, in the understanding and prediction of human behavior are deep, real, and difficult. This study tested two conflicting sets of predictions concerning divorce patterns in the United States: a socioeconomic set and a biocultural set. The socio-economic perspective predicts that the addition of one or more children to a marriage would increase the husband's (as compared with the wife's) motivation to petition for a divorce. The biocultural perspective posits that a man-to-child affiliative bond would add to a man's adherence to his child(ren) and, in turn, would lower his motivation (as compared with the woman's) to petition for a divorce. The biocultural perspective was supported in this study. PMID- 8245910 TI - Children's perceptions of others' kindness in helping: the endocentric motivations of pride and guilt. AB - Elementary school and college students rated the kindness of helping by story protagonists with different attributed motivations. Of particular interest was the effect of the endocentric (self-serving) motivations of anticipated pride and guilt on the kindness ratings. A number of prosocial theorists view such endocentrically motivated helping as less altruistic than exocentrically (other serving) motivated helping. Compared with helping attributed to the exocentric motivation of sympathy, helping attributed to guilt avoidance led to lower ratings of kindness by all but second graders. Pride-attributed helping, however, did not result in lower kindness ratings at any grade level. The motivational attributions of praise and reward attainment and criticism and punishment avoidance led to relatively low kindness ratings, with the two avoidance motivations leading to the lowest ratings. The latter finding suggests an alternative explanation of the kindness ratings for guilt-motivated helping. PMID- 8245911 TI - Preschool children's decoding of facial emotions. AB - This study examined preschool children's ability to decode facial emotions. Subjects comprised 32 preschoolers (16 3 1/2-year-olds, 16 5-year-olds), with equal numbers of boys and girls, enrolled in a preschool. Children heard a brief story describing a boy's emotion and were shown three photographs of a boy, each displaying a different emotion (one target emotion and two distractor or nontarget emotions). Children were asked to choose the one photograph that corresponded to the boy's emotion in the story. As predicted, the ability to decode facial emotions improved with age for both boys and girls. Girls were significantly better than boys at identifying emotions; in fact, 3 1/2-year-old girls were as accurate as 5-year-old boys. Age and gender effects, as well as topics for future investigation, are discussed. PMID- 8245912 TI - Understanding the combination of memory variables: a re-examination. AB - This study was a direct prolongation of a study by Wellman, Collins, and Glieberman (1981), which showed that, provided a sufficiently sensitive methodology is used, it is possible to demonstrate experimentally that even 5 year-old children are capable of integrating two types of information--effort and difficulty of task--in their judgment of performance (number of items recalled). We prolonged this study by adopting Anderson's information integration methodology in order to provide evidence concerning the type of combinatorial rule describing subjects' use of both pieces of information and to refine analysis to the individual level. In terms of group results, it was possible to describe the algebraic aspects of the information integration process by an additive model in children and adolescents, and by a multiplicative model in adults. Conclusions changed considerably, however, when individual results were studied. In the 5- and 8-year-old children, the dominant rule was clearly unifactorial, involving effort. Only in the adult group was true integration seen in all subjects. PMID- 8245913 TI - The place of emotion in stories told by children: an exploratory study. AB - This study explored the relationship between developmental changes in children's understanding of emotions and their use of affective content (Wilkinson, Barnsley, Hanna, & Swan, 1980) and affective structure (Brewer & Lichtenstein, 1982) in the production of stories. We hypothesized that children with an advanced understanding of emotions would make more use of both affective content and affective structure. This relationship was found only for affective content, which was more advanced in girls than boys. Affective structure did not relate to affective understanding or sex but, rather, to verbal intelligence. The implications of these results for the part played by psychological processes in children's story telling are discussed. PMID- 8245914 TI - Are individual differences in fantasy play related to peer acceptance levels? AB - Individual differences in fantasy play, as they relate to cognitive competence and peer acceptance, were examined in a sample of 66 children between the ages of 4 and 8 years. Each child's free-play behaviors were videotaped and then coded for frequency of fantasy play and nonfantasy play, as well as for the unreality level and unfamiliarity level of the fantasy play. Solitary play was rarely observed; therefore all play measures that were coded were social (i.e., play involving at least one partner). Cognitive competence measures included self ratings and teacher ratings of academic skills. Peer acceptance measures included self-ratings and teacher ratings of friendships and social activities. Results showed that high-fantasy players had higher self-ratings of peer acceptance than did average-fantasy players. High-fantasy players, however, had lower teacher ratings of peer acceptance than did average-fantasy players. These differential relations could be related to an inability on the part of high-fantasy players to distinguish imagined popularity from actual peer acceptance levels. Future research is recommended to provide more detailed characterizations of individual differences in fantasy play and how they relate to outcomes in the social, cognitive, and affective domains. PMID- 8245915 TI - Does perceptual intake speed reflect intelligent use of feedback in an inspection time task? The effect of restricted feedback. AB - To examine whether the development of effective strategies during inspection time (IT) required active learning processes, we gave subjects an IT task involving false feedback. This IT task (ADIT) started at an exposure duration of 20 ms and gradually increased until subjects could reliably discriminate IT stimuli. False feedback about correctness of discrimination was introduced to make the ADIT task particularly difficult for individuals who were attempting to develop and refine an IT-related strategy. There was no difference between ITs derived from the ADIT and a standard IT task (VIT). There were no differences in ITs for subjects given either truthful or false feedback on the ADIT task. However, an interaction did exist between the feedback condition and the two IT tasks. This interaction indicated that subjects with ostensibly better VITs had poorer performance on ADIT, compared with those subjects who had truthful feedback. This finding suggests that false feedback can disrupt effective performance on IT. Self reported strategy use had a significant independent effect on the observed IT measures. Individuals who reported strategies were not significantly higher in IQ than those who did not report strategies. PMID- 8245916 TI - The relationship of worrying, coping, and symptoms among college men and women. AB - College students maintained a daily log of their worrying for 7 consecutive days, with three different subgroups starting on different days of the week. Patterns of the type and amount of worrying did not vary systematically over successive days. Multiple regression analyses showed a direct relationship between worrying level and amount of psychological symptoms, as measured by both the diary method and a single item self-rating. Avoidance coping was related to higher levels of physical and psychological symptoms, whereas problem solving was associated with fewer psychological symptoms for females. Men and women, for the most part, had similar relationships among the variables. PMID- 8245917 TI - Antibody response to rat erythrocytes and some behaviors in two strains of mice after social isolation. AB - The purpose of this research was twofold: (a) To find out if the antibody response and some behaviors are differential traits for mice (murine strains C57BL/6 and BALB/c); and (b) to study the influence of strain and isolation on both behavior and the antibody response. We used 21 C57BL/6 male mice and 28 BALB/c male mice; of these, 11 C57BL/6 and 16 BALB/c were kept isolated. The rest were housed 2 mice to a cage. We used three open-field tests (of varying light and sound stimulation), a holeboard test, and a light-darkness test. All the mice were immunized with rat erythrocytes. In addition, the mice were weighed. The traits that best differentiated both strains were weight, IgM primary response, and emotional behavior (behaviors in stimulating open fields). Social isolation (of adult mice) influenced emotional behavior, but not the antibody response. PMID- 8245918 TI - Detecting slant-in-depth of real trapezoidal and rectangular surfaces: moving monocular viewing equivalent to stationary-binocular viewing. AB - Cues from binocularity and observer motion are often believed to be more important in perceiving depth than pictorial cues such as relative visual size and linear perspective. Both binocularity and motion are effective in simulated displays. However, for real stimuli evincing nonveridical pictorial cues, binocularity has been more effective than motion; sometimes motion has had an insignificant effect. This may reflect inadequate extent of motion, an assertion investigated in the present study. Two groups of observers determined whether rectangular and trapezoidal surfaces were slanted-in-depth under stationary monocular (SM), stationary-binocular (SB), and moving-monocular conditions with 15-cm (15MM) and 25-cm (25MM) lateral head-motion extents according to group. The trapezoidal surfaces appeared as rectangular during SM viewing to mislead regarding slant. The effect of pictorial cues was substantially diminished during SB viewing whereas 15MM viewing was weak, 25MM was as effective as SB viewing. Comparison of the overall numbers of correct responses for the two groups indicated no contextual biasing. PMID- 8245919 TI - Functional and dysfunctional impulsivity and depression and suicidal ideation in a subclinical population. PMID- 8245920 TI - Environmental factors related to the outcome of panic disorder. A seven-year follow-up study. AB - The purpose of this study was to examine factors related to the outcome of naturalistically treated panic disorder. In order to achieve this we followed up 69 patients 7 years after they had presented at a psychiatric clinic. At follow up, the patients were generally doing well despite persisting symptoms. Patients who were more severely ill at the time of initial assessment had a worse outcome. These patients had more severe panic and agoraphobic symptoms, had illnesses of longer duration, and more often had histories of major depression. Among the developmental variables examined, separation from a parent by death or divorce was strongly related to poor outcome. Other factors associated with poor outcome included high interpersonal sensitivity, low social class, and unmarried marital status. The findings show that, for this chronic illness, measures of severity and chronicity predict more severe and persisting symptoms. They also indicate that outcome is importantly related to the social environment in which the illness develops and with which it interacts. PMID- 8245921 TI - Panic and prolapse. Meta-analysis. AB - Studies concerning the association of panic disorder and mitral valve prolapse have produced mixed results. Meta-analysis was used to quantitatively review the literature and explain conflicting results. Only 19 published and two unpublished studies have documented both panic and mitral valve prolapse in addition to including a control group, and were therefore included. The strength of association between panic disorder and mitral valve prolapse was measured as the odds ratio and study variables were included in the analysis. The overall weighted relative risk was 2.3 (confidence interval, 1.6 to 3.5). Important study variables were often not addressed in the literature and stepwise multiple regression found that auscultatory criteria, control group diagnosis, and sample size predicted relative risk (F = 11.34, p < .001). p-Values differed with publication status (t = 2.32, p = .033). Although the quality of literature is uneven, there appears to be a significant association between panic disorder and mitral valve prolapse. However, the suggestion of publication bias indicates that the published literature may present a biased view. PMID- 8245922 TI - Traumatic stress of a wartime mortuary. Anticipation of exposure to mass death. AB - Exposure to traumatic death is a risk factor for posttraumatic stress disorder. For some groups, anticipation of such exposure may contribute to traumatic stress. We studied the anticipated stress of working in the Operation Desert Storm mortuary in two groups prior to the arrival of the dead. We examined those who would handle remains (mortuary workers, N = 386, 330 men and 56 women) and those who would not (support workers, N = 87, 67 men and 20 women). These two groups were a mixture of volunteers and nonvolunteers as well as persons with and without experience in handling the dead. The mortuary workers had higher levels of preexposure distress than support workers; nonvolunteer mortuary workers had higher levels of distress than volunteers. Female mortuary workers had higher levels of distress than males, although this effect was modest. Experienced mortuary workers reported fewer intrusive and avoidant symptoms than did inexperienced workers. Those persons at highest risk for generalized distress as well as intrusive and avoidant symptoms were inexperienced nonvolunteer mortuary workers. PMID- 8245923 TI - The role of the helping alliance in psychiatric community care. A prospective study. AB - In this study, we assessed the quality of the helping alliance between patients and clinical case managers in psychiatric community care and examined its value as predictor of treatment outcome. Patients were interviewed about five different aspects of the helping alliance using simple questions and visual analogue scales. The duration and degree of hospitalization and changes in the patients' working and accommodation situations during a 20-month follow-up period were obtained as outcome criteria for 72 patients with mostly psychotic disorders receiving long-term treatment. The patients' general view of the helping alliance was quite positive. Some aspects of the helping alliance were significantly correlated with hospitalization and changes in working situation during the follow-up period, which indicated a better outcome for patients who experienced the helping alliance more positively. The findings suggest that the helping alliance may be a relevant therapeutic factor not only in psychotherapy, but also in complex psychiatric treatment settings. PMID- 8245924 TI - Rating depression in schizophrenia. A comparison of a self-report and an observer report scale. AB - Prior research has indicated an inconsistent agreement between self-report and observer report of depression in schizophrenia. The purpose of this study was to assess the level of agreement between a self-report measure of depression and a structured interview. Both outpatients (N = 100) and inpatients (N = 50) with schizophrenia were assessed using the Beck Depression Inventory and a structured interview scale, the Calgary Depression Scale. Scores were convergent. However, a sizable proportion of inpatients had difficulty completing the self-report instrument. It is concluded that depressed affect can be assessed in schizophrenics by both self-report and structured interview, but the Beck Depression Inventory poses difficulties in use with inpatients. PMID- 8245925 TI - Interictal violence in epilepsy. Relationship to behavior and seizure variables. AB - Interictal violence among epileptic patients could result from factors other than epileptiform activity. We characterized 44 patients who presented for psychiatric evaluation because of violent behavior. Most violent acts consisted of verbal or minor physical aggression. Twenty (45%) of these patients met criteria for a schizophrenic disorder, and one committed murder during a paranoid schizophrenic relapse. In addition to schizophrenia, the violence patients had significantly more mental retardation when compared with 88 age- and sex-matched epileptic patients without prior violent behavior. However, violent and nonviolent patients did not differ on seizure variables such as type and frequency of seizures, auras, electroencephalographic changes, epilepsy age of onset, or anticonvulsant therapy. These findings suggest that interictal violence is associated more with psychopathology and mental retardation than with epileptiform activity or other seizure variables. PMID- 8245926 TI - Men with sexual interest in transvestites, transsexuals, and she-males. AB - This study was a preliminary exploration of gynandromorphophilia, that is, sexual interest in cross-dressed or anatomically feminized men. Subjects were male subscribers to a voice mail system devoted to personal advertisements for sexual or romantic partners. These comprised 51 gynandromorphophiles who sought cross dressers, transvestites, transsexuals, or she-males for such relationships, 37 gynandromorphophilic cross-dressers who identified themselves as cross-dressers and sought similar men, and 31 residual cross-dressers who sought masculine or unspecified male partners. Analysis of advertisement content suggested that gynandromorphophilia constitutes a distinct erotic interest. PMID- 8245927 TI - Do substance abuse patients with more psychopathology receive more treatment? AB - The association between psychopathology at treatment entry and the amount of treatment services received was evaluated in 104 alcohol-dependent and 100 cocaine-dependent male veterans treated for 1 month in either a day hospital or inpatient program. Measures of psychopathology included the Addiction Severity Index psychiatric composite score, the presence or absence of an antisocial personality disorder diagnosis, and the total number of additional lifetime or current psychiatric diagnoses. Patients with higher admission Addiction Severity Index psychiatric composite scores received more medical, alcohol, family/social, and psychiatric services. There was also preliminary evidence that patients who received more treatment showed greater improvement 7 months after admission. The relationships between the other measures of psychopathology and treatment services failed to achieve overall statistical significance, although significant relationships were found in several individual areas. PMID- 8245928 TI - Discrimination of facial emotion in depressed patients with visual-perceptual disturbances. PMID- 8245929 TI - The triply diagnosed: patients with major mental illness, cognitive impairment, and substance abuse. PMID- 8245930 TI - Ear stuffing: an unusual form of self-mutilation. PMID- 8245931 TI - Substance abuse and impulse control. PMID- 8245932 TI - Annual emergence patterns of Culex nigripalpus females before, during and after a widespread St. Louis encephalitis epidemic in south Florida. AB - Resting Culex nigripalpus females were collected from the same site at least 3 times a week from January 1986 through December 1991. Newly emerged mosquitoes were used to monitor annual emergence patterns that were rainfall driven. Four of the years, 1986-89, preceded the 1990 St. Louis encephalitis (SLE) epidemic in south Florida. The pre-epidemic period averaged 4 emergences per year, with most occurring during July and August. The emergence patterns observed during the epidemic year were unusual in their early start in May and June and in the total number (11) observed. Adult mosquito emergences in 1991 were similar to those in 1990, with an early start and a total of 14 observed, yet no SLE transmission was recorded in Indian River County during 1991. A time series analysis of the 6-year data set showed a significant cross-correlation between the emergence of Cx. nigripalpus females and heavy (> 50 mm) rains. A significant adult emergence occurred 5-8 days and 12-15 days following a heavy rainfall event. PMID- 8245933 TI - In vitro assay for permethrin persistence and interference with bloodfeeding of Culicoides (Diptera: Ceratopogonidae) on animals. AB - Permethrin (65% AI) was applied to the dorsum of 3 goats (3-4 ml/animal). Hair clippings were taken before treatment and 4-6, 18-20, 39-41 and 67-69 days after treatment. Culicoides variipennis fed upward through a thin layer of hair and through a membrane on an artificial feeder containing cattle blood. Midge knockdown was usually complete after the 45 min exposure period, but many bloodfed first. Belly hair residues did not statistically reduce engorgement beyond 4-6 days. Head hair residues reduced engorgement by up to 98% at 18-20 days, but did not significantly reduce engorgement at 67-69 days. Back hair residues significantly reduced engorgement for the entire period (99-100% through day 39-41 and 66% by day 67-69). Partial engorgement was higher in the treatments. PMID- 8245934 TI - Evaluation of light traps for sampling anopheline mosquitoes in Kilifi, Kenya. AB - Anopheline mosquitoes were sampled inside houses, where residents slept under untreated bednets, by CDC light traps and human-biting catches to evaluate light traps as a means for determining human exposure to malaria vectors in Kilifi District, Kenya. Mosquitoes were sampled during 2 all-night collections by light traps and one all-night biting catch in a series of 262 houses. Collections yielded 1,721 Anopheles gambiae s.l. and 46 An. funestus, and 60.3% of the houses were negative for anophelines. There was a significant correlation in numbers of An. gambiae s.l. captured by light traps and human-biting collections (r = 0.64), but light traps were biased and underestimated An. gambiae s.l. abundance. This bias increased with increasing mosquito abundance. In addition, the proportion of An. gambiae s.l. infected by Plasmodium falciparum was 2.3-fold higher in light traps than in human-biting collections. Along the coastal zone of Kenya where vector abundance is low, light traps do not provide an adequate estimate of man vector contact when such information is required at the household level in epidemiological studies of malaria parasite transmission. PMID- 8245935 TI - Insect blood meal studies using radiosodium 24Na and 22Na. AB - Rabbits were infused with 3.7 GBq (100 mCi) of [24Na]Na ion in a 100-ml sodium carbonate solution. Beta particles were detected using a Tennelec Counting System; background counts were 1.6 +/- 1 counts per minute (cpm). Counts for one nanoliter of blood ranged from 22 to 30 cpm. Blood volumes on the mouthparts of tabanids following a 15-sec interrupted feeding were estimated to be 12.5 nl for Tabanus fuscicostatus, 10.8 nl for T. nigrovittatus and 6.12 nl for Chrysops fuliginosus. Estimates of the quantity of blood adhering to 22-gauge needles and insect pins (size 2) following a percutaneous intramuscular needle-stick were 8.8 +/- 1.0 nl and 5.7 +/- 1.8 nl, respectively. Mosquitoes, Aedes aegypti females, were fed to repletion with a 22Na-artificial diet, and radioactivity was measured using a Packard Autogamma 5650. The estimated average blood meal size was 2.80 +/ 0.94 microliters. PMID- 8245936 TI - Chironomid (Diptera: Chironomidae) larval occurrence and transport in a municipal water system. AB - Chironomid larval populations contaminating a municipal water system were monitored over a 24-month period between June 1987 and June 1989. Larval drift within distribution lines occurred throughout the year, and larval suspension was uniform at all depths within open reservoirs. Draining and cleaning reservoirs to rid them of chironomid populations was of questionable value because larval populations recovered fully within weeks, and larval drift occurred throughout the system. PMID- 8245937 TI - Laboratory evaluation of the irritancy of bendiocarb, lambda-cyhalothrin and DDT to Anopheles gambiae. AB - In a laboratory study, the irritancy of bendiocarb, lambda-cyhalothrin and DDT to Anopheles gambiae was evaluated at field, 1/3 field and 1/10 field rates using WHO conical exposure chambers and excito-repellency test boxes. Bendiocarb was the least irritant insecticide at all rates, inducing levels of takeoff, flight and exiting behavior similar to those of a distilled water control treatment. Of those mosquitoes introduced to the bendiocarb-treated boxes, not more than 1% exited and survived at any dose rate. Lambda-cyhalothrin and DDT were highly irritant to An. gambiae, inducing a strong stimulation to take off and fly and also a high level of exiting. Exiting-survival rates associated with lambda cyhalothrin and DDT were between 15 and 51%. The relevance of these findings to the control of mosquito populations and the prevention of malaria transmission is discussed. PMID- 8245938 TI - A new cytotype of Anopheles nuneztovari from western Venezuela and Colombia. AB - Cytogenetic analysis of the larval polytene chromosomes of Anopheles nuneztovari from 5 collection sites in Tachira and Zulia states northwest of the Andean Cordillera in western Venezuela and from 2 sites in the Department of Valle, western Colombia, revealed what appears to be a distinctive cytotype informally designated as An. nuneztovari C. Its chromosomes are homosequential with those of An. nuneztovari B from western Venezuela southeast of the Cordillera but differ in the presence of a well-defined chromocenter and unique inversion polymorphisms. The large complex inversion in western Venezuela, 2Lb, is present at a frequency of 0.263 and deviates significantly from Hardy-Weinberg equilibrium in 3 of the 5 sites. Two smaller inversions (2Lc and 2Ld) that are included in 2Lb are present in the Colombian samples at a frequency of 0.300. PMID- 8245939 TI - Efficacy of two formulations of the insect growth regulator, pyriproxyfen (Nylar or Sumilarv), against nuisance Chironomidae (Diptera) in man-made ponds. AB - A juvenoid insect growth regulator, pyriproxyfen (Nylar or Sumilarv), in a 10% emulsifiable concentrate (10 EC) and a granular (3% sand granules) formulation was evaluated for efficacy against natural populations of chironomid midges in man-made ponds in central Florida. Both formulations at 0.05 kg AI/ha induced complete suppression of adult midge emergence for 1 wk. At 0.2 kg AI/ha, emergence of adult midges completely ceased for 2 and 5 wk with the EC and the granules, respectively. The EC formulation was not effective beyond 2 wk, but the granules at both treatment rates reduced midge emergence by 81-100% for 9 wk posttreatment. The differences in suppression of adult midge emergence between the 2 formulations and times posttreatment were both highly significant (P < 0.01). No significant difference (P > 0.05) was found between the posttreatment percent inhibition of midge emergence caused by the lower and the higher rates of the granular formulation. Some reductions of midge larvae occurred in the ponds receiving the higher rate of the 2 formulations, but these reductions were statistically not significant. PMID- 8245940 TI - Photoperiod and longevity in Anopheles crucians. AB - The effect of photoperiod on longevity of nondiapausing members of Anopheles crucians was evaluated in laboratory experiments. First-generation adults reared from field collections illustrated the same trends. Individuals reared and maintained under short photoperiod (either 8 h light:16 h dark or 11 h light:13 h dark) lived longer than did those reared and maintained under long photoperiod (either 16 h light:8 h dark or 15 h light:9 h dark). Analysis of covariance showed that the greater longevity of short-photoperiod mosquitoes was not caused by their larger average body size. Also, a paired t-test confirmed the effect of photoperiod, independent of body size, on longevity; in mosquitoes paired according to thorax length, short-photoperiod individuals lived longer than did long-photoperiod individuals of the same sex and cohort. The reason short photoperiod mosquitoes of this species live longer is not known. PMID- 8245941 TI - Inheritance and linkage of malic enzyme in Anopheles stephensi. AB - Genetics and linkage analysis of malic enzyme (Me) have been worked out in Anopheles stephensi. Genetic analysis revealed the 2 variants to be codominant alleles at a locus Me, which is sex-linked. Linkage studies with another X-linked mutant red-eye (r), indicated that the map distance between 2 loci was 44.60 +/- 1.07. PMID- 8245942 TI - Comparison of sampling efficacy of sweeping and dipping for Aedes aegypti larvae in tires. AB - A sweep net with a flexible plastic frame was compared to a dipper for sampling efficacy for Aedes aegypti larvae in different-sized tires. When known numbers of mosquito larvae were introduced into tires, from 2 to 6.8 times more larvae were recovered by sweeping than by dipping. Sediment in the tires significantly reduced the recovery of larvae by both methods. Sampling efficacy was affected by water level and tire size. In relation to dipping, sweeping showed less variation in percentage recovery, higher sensitivity for detecting larvae and had better correlations with known numbers of larvae in tires. We conclude that sweeping is superior to dipping for sampling mosquito larvae in tires. PMID- 8245943 TI - Intradomiciliary behavior of Anopheles albimanus on the coastal plain of southern Mexico: implications for malaria control. AB - The postfeeding indoor resting behavior of Anopheles albimanus in experimental houses in southern Mexico was investigated by using a mark-recapture procedure. The majority of mosquitoes rested inside houses after taking a blood meal indoors. There was a higher landing frequency on interior surfaces other than walls and roofs; however, mosquitoes rested for longer periods on these 2 surfaces. Successive landings on walls after short flights showed that mosquitoes gradually increased their mean landing height from 1.0 to 1.4 m. Similarly, mosquitoes resting at the base of inner roofs had a successive landing height range of about 0.5 m. Based on these observations and the potential for reduction of nearly 50% in the quantity of insecticide used and the time needed to apply it, village-scale studies involving the selective spraying of a 1-m-wide swath of insecticide on walls and on roofs are recommended in this area. PMID- 8245944 TI - Field trials of Bacillus thuringiensis H-14 and Bacillus sphaericus (strain 2362) formulations against Anopheles arabiensis in the central highlands of Madagascar. AB - Malaria is highly endemic and unstable in the central Highlands plateau of Madagascar. The infection is seasonally transmitted by Anopheles funestus and An. arabiensis. The latter species is abundant especially in rice-growing areas. The field efficacies of commercial formulations of Bacillus thuringiensis H-14 and B. sphaericus (strain 2362) were assessed against An. arabiensis in 5 types of larval habitats. The granular formulation of B. thuringiensis (Vectobac GR) provided very good control in small ponds and rainwater ditches. In ricefields complete larval control was achieved with Vectobac GR and the flowable concentrate (Vectobac 12AS) at 2.5 kg/ha and 0.6 liter/ha, respectively. The granular formulation of B. sphaericus (ABG 6185) showed activity similar to that of Vectobac GR in small pools and ricefields. ABG 6185 was less effective in rainwater ditches where it gave satisfactory control at rates not lower than 6 kg/ha. PMID- 8245945 TI - Use of cellular fatty acid analysis to characterize commercial brands of Bacillus thuringiensis var. israelensis. AB - The cellular fatty acid composition of Bacillus thuringiensis var. israelensis (B.t.i.) from 5 commercial brands (Vectobac, Acrobe, Skeetal, Bactimos and Teknar), as well as of the current International Standard for B.t.i. (IPS 82), was determined using a Hewlett-Packard Microbial Identification System. The original strain of B.t.i., B.t. var. kurstaki, B.t. var. thuringiensis, B.t. var. morrisoni and Bacillus sphaericus strain 2362 were used as outgroups. Acrobe, Bactimos, Teknar, Vectobac and IPS 82 consisted of the same strain. Skeetal represented a different strain than the other commercially produced B.t.i. Our results indicate that cellular fatty acid analysis can be used to distinguish among the forms of B.t.i. produced by various manufacturers. PMID- 8245946 TI - Field trial of two Bacillus thuringiensis var. israelensis formulations for control of Aedes species mosquitoes in Michigan woodlands. AB - Vectobac and Bactimos corn cob granules were evaluated for control of Aedes species mosquito larvae in woodland pools. No significant differences were seen between the 2 formulations. Both provided greater than 90% control at application rates as low as 100 mg/m2 (0.89 lb/acre) and greater than 98% control at label specified field application rates (2.5 or 5 lb/acre). PMID- 8245947 TI - Oviposition response of Aedes aegypti mosquitoes to different concentrations of hay infusion in Trinidad, West Indies. AB - Ovitraps containing various concentrations of hay infusion and tap water were exposed weekly in the field for 15 wk to determine the oviposition patterns of Aedes aegypti. The results showed 10, 20, 60 and 80% hay infusions each attracted similar numbers of Ae. aegypti eggs oviposited and egg occurrences. No repellent effect was observed. In another field study, significantly more eggs and egg occurrences were collected from 25 and 50% hay infusions and tap water. The differences in these results from those of a previous study in Puerto Rico are discussed. PMID- 8245948 TI - Distributional records for Aedes mosquitoes from the U.S. Air Force ovitrapping program-1992. AB - During 1992, ovitrapping to sample populations of container-breeding Aedes was conducted at 28 U.S. Air Force installations in North America. Aedes albopictus was collected at 19 installations. The collections of Ae. albopictus at Goodfellow Air Force base (AFB), TX, and Moody AFB, GA, represent new records for Tom Green and Lowndes counties, respectively. Six installations were positive for Ae. aegypti; 13 were positive for Aedes triseriatus. PMID- 8245949 TI - Predation by a cyprinodontid fish, Aphanius mento, on Culex pipiens: effects of alternative prey and vegetation. AB - We assessed the effects of an alternative prey species (Daphnia magna) and a submergent plant (Ceratophyllum demersum) on the efficiency of the fish, Aphanius mento, to prey on mosquito (Culex pipiens) larvae in the laboratory. Aphanius mento is an untested biological control candidate of mosquitoes. When D. magna was introduced with equal numbers of Cx. pipiens larvae, there was a 50.7% reduction in predation by A. mento on the mosquito larvae. However, the more D. magna a fish ate, the more Cx. pipiens it consumed. Ceratophyllum demersum did not affect predation rates on Cx. pipiens larvae. PMID- 8245950 TI - Mosquito repellent action of neem (Azadirachta indica) oil. AB - Two percent neem oil mixed in coconut oil, when applied to the exposed body parts of human volunteers, provided complete protection for 12 h from the bites of all anopheline species. Application of neem oil is safe and can be used for protection from malaria in endemic countries. PMID- 8245951 TI - Neem oil as a sand fly (Diptera: Psychodidae) repellent. AB - The repellent action of neem oil was evaluated against sand flies under laboratory and field conditions. Concentrations of 2% neem oil mixed in coconut or mustard oil provided 100% protection against Phlebotomus argentipes throughout the night under field conditions; against Phlebotomus papatasi it repelled sand flies for about 7 h in the laboratory. Neem oil is an indigenous product and a low-cost alternative for personal protection against sand fly bites. PMID- 8245952 TI - Distribution, abundance and bionomics of Aedes albopictus in southern Texas. AB - A survey was conducted for Aedes albopictus in southern Texas during the summer of 1992. Thirty-five new country records were added to the distribution of this imported mosquito in Texas. Aedes albopictus was widely distributed throughout the ecological regions in the survey area, but its abundance decreased in counties adjacent to the Rio Grande River. However, these counties had high densities of Aedes aegypti. PMID- 8245953 TI - Binding of plasma selenoprotein P to cell membranes. AB - Competitive binding assays were performed to determine the amount of binding of 75Se-labeled plasma selenoprotein P (PSP) to membranes from different rat tissues at physiologic pH. 75Se PSP for use as a ligand in the binding assays was labeled in vivo by injecting rats with 75Se selenious acid. PSP was obtained from plasma by salt precipitation and affinity and ion-exchange chromatography. Membranes for receptor assays were prepared from liver, kidney, testes, and brain of rats fed diets with either 0.01, 0.1, or 2 ppm selenium. At pH 7.4 PSP was bound differentially to tissues in the following order: brain > kidney > testes > liver. Specific binding of PSP to tissue membranes from rats fed the different levels of selenium increased with increasing amounts of dietary selenium. Saturation assays indicated apparent saturation of membrane receptors by the 75Se labeled PSP. Another significant new finding was a 134-kDa complex of PSP and membrane receptor, identified by gel-filtration chromatography of cross-linked samples from binding assays. This provides evidence for a membrane receptor for PSP in rat tissues. PMID- 8245954 TI - Carbethoxylation of coordinated histidine by diethylpyrocarbonate. AB - The metastable, reversible carbethoxylation of histidine in the cobalt(III) complexes (ethylene-diamine)(histidine)chlorocobalt(III) chloride, [Co(III)(en)ClHis]Cl, and (diethylenetriamine) (histidine)cobalt(III) dichloride, [Co(III)(dien)His]Cl2, following reaction with diethylpyrocarbonate, was observed using UV spectroscopy. These observations indicate that diethylpyrocarbonate can react with a coordinated imidazole ring. CD transitions associated with coordinated carbethoxyhistidine in [Co(III)(en)ClHis]Cl and [Co(III)(dien)His]Cl2 were also observed. The molar ellipticities of the CD bands observed for carbethoxy [Co(III)(en)ClHis]Cl or carbethoxy [Co(III) (dien)His]Cl2 are much larger than the molar ellipticities of the CD transition associated with carbethoxy N alpha-acetyl-L-histidine, indicating that loss of rotational freedom of the imidazole ring could be a major factor in the enhanced CD. PMID- 8245955 TI - Further characterization of the redox and spectroscopic properties of Azotobacter vinelandii ferritin. AB - Bacterial ferritin from Azotobacter vinelandii (AVBF) has many properties in common with and a number of properties distinct from the more thoroughly studied animal ferritins. The most notable differences are the high phosphate content of the mineral core and the presence of heme (12 per AVBF) in AVBF. In both ferritin types, redox reactions are essential to the iron release and deposition function of the ferritins. The heme reduction potential in apo AVBF is pH independent as are both the heme and core reduction potential in holo AVBF. pH measurements confirm the pH independence for heme reduction in apo AVBF; however, they establish the conflicting result that 1.7 +/- 0.2 protons per iron atom are taken up during core reduction. These results are interpreted as a two-step reduction process consisting of a pH independent reduction of heme in holo AVBF followed by a pH dependent reduction of the mineral core. Detailed spectroscopic studies have been undertaken to determine if heme-core interactions are detectable during the redox reactions of AVBF. Optical spectroscopy of the heme groups in apo AVBF demonstrate that all twelve are identical and undergo uniform and rapid reduction. EPR spectroscopy establishes the presence of both low-symmetry, g = 4.3, Fe3+ from the mineral core and low-spin heme with g values of 2.87, 2.32, and 1.46 in holo and identical g values for the low-spin heme in apo AVBF. EPR integration of the heme groups in both apo and holo gave values of 13.2 +/- 1.3 heme spins per AVBF at 4.2, 10, 25, 35, and 45 K. No heme perturbations were detected in holo or apo AVBF by Resonance Raman and circular dichroism spectroscopy. Both reduced and oxidized apo AVBF gave normal fluorescence emission at 330-340 nm when excited at 279 nm. These spectroscopic, redox, and reactivity results provide more detailed properties of AVBF for comparison with other bacterial and animal ferritins. PMID- 8245956 TI - [125I]RTI-55 binding to cocaine-sensitive dopaminergic and serotonergic uptake sites in the human brain. AB - [125I]RTI-55 is a newly synthesized cocaine congener that may offer advantages over other ligands previously used to examine cocaine binding sites. However, the in vitro pharmacological and anatomical characterization of [125I]RTI-55 binding sites has not been previously performed in human brain. To determine the specificity, stability, and feasibility of [125I]RTI-55 for use in radioligand binding assays in postmortem human tissue, a series of experiments were performed characterizing [125I]RTI-55 binding sites in human brain using homogenized membrane preparations and quantitative autoradiography. Analysis of the association, dissociation, and saturation data favored two-phase processes. A curve-fitting analysis of the data derived in saturation experiments found a high affinity site with KD = 66 +/- 35 pM and Bmax = 13.2 +/- 10.1 pmol/g of tissue and a low-affinity site with KD = 1.52 +/- 0.55 nM and Bmax of 47.5 +/- 11.2 pmol/g of tissue. Competition by ligands known to bind to the dopamine transporter showed a rank order of RTI-55 > GBR-12909 > maxindol > WIN 35428 > = methylphenidate > (-)-cocaine > buproprion > (+)-amphetamine. Binding to serotonergic sites was evaluated in the midbrain. Results of the saturation experiment performed autoradiographically in the midbrain showed a single site with KD = 370 +/- 84 pM. It appears that [125I]RTI-55 should be useful in further studies of the regulation of cocaine binding sites using postmortem human specimens. PMID- 8245957 TI - Brain alpha-ketoglutarate dehydrogenase complex activity in Alzheimer's disease. AB - We measured the activity of the alpha-ketoglutarate dehydrogenase complex (alpha KGDHC), a rate-limiting Krebs cycle enzyme, in postmortem brain samples from 38 controls and 30 neuropathologically confirmed Alzheimer's disease (AD) cases, in both the presence and absence of thiamine pyrophosphate (TPP), the enzyme's cofactor. Statistically significant correlations between brain pH and lactate levels and alpha-KGDHC activity in the controls were observed, suggesting an influence of agonal status on the activity of alpha-KGDHC. As compared with the controls, mean alpha-KGDHC activity, with added TPP, was significantly (p < 0.005) reduced in AD brain in frontal (-56%), temporal (-60%), and parietal ( 68%) cortices, with the reductions (-25 to -53%) in the occipital cortex, hippocampus, amygdala, and caudate failing to reach statistical significance. In the absence of exogenously administered TPP, mean alpha-KGDHC activity was reduced to a slightly greater extent in all seven AD brain areas (-39 to -83%), with the reductions now reaching statistical significance in the four cerebral cortical areas and hippocampus. A statistically significant negative correlation was observed between alpha-KGDHC activity and neurofibrillary tangle count in AD parietal cortex, the brain area exhibiting the most marked reduction in enzyme activity; this suggests that the enzyme activity reduction in AD brain may be related to the disease process and severity. In each brain area examined, TPP produced a greater stimulatory effect on alpha-KGDHC activity in the AD group (23 280% mean stimulation) as compared with the controls (-4 to +50%); this TPP effect could be explained by reduced endogenous TPP levels in AD brain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245958 TI - Immunochemical characterization of the beta 2 subunit of the GABAA receptor. AB - To date three beta subunits of the GABAA receptor have been identified in rat brain as a result of cDNA library screening. The beta 2 subunit has been reported to have a wide distribution in rat brain based on in situ hybridization studies quantifying beta 2 mRNA. To study the beta 2 subunit more directly, we have raised a polyclonal antibody to a synthetic peptide representing residues 315-334 of the intracellular loop of the beta 2 subunit. The antibody, which had been affinity-purified, recognized the beta 2 peptide but did not immunolabel homologous beta 1 and beta 3 subunit peptides, indicating that this antibody is specific for the beta 2 subunit of the receptor. In western blots of the purified receptor, the antibody recognized a major diffuse band of 54-58 kDa and exhibited minor labeling of lower-molecular-mass polypeptides. In western blots of cortex homogenate, the antibody exhibited nervous system-specific labeling of a 55-kDa band that comigrated with the 55-kDa band of the purified receptor. Quantitative immunolabeling of this 55-kDa polypeptide permitted direct determination of the relative amounts of the beta 2 subunit in different brain regions. The brainstem contained the highest relative specific activity of the beta 2 subunit, followed by the inferior colliculus, olfactory lobe, and cerebellum. Lower levels of immunolabeling were seen in hypothalamus, hippocampus, thalamus, and cortex. PMID- 8245959 TI - Regional and interspecies differences in brain progesterone metabolism. AB - Metabolites of [3H]progesterone were studied in slices prepared from different brain regions of male rat, mouse, and monkey. The major metabolites were 5 alpha dihydroprogesterone (5 alpha-DHP) and 3 alpha,5 alpha-tetrahydroprogesterone (3 alpha,5 alpha-THP) in rat brain slices, 5 alpha-DHP and 20 alpha dihydroprogesterone (20 alpha-DHP) in mouse brain slices, and 20 alpha-DHP in monkey brain slices. In rat olfactory bulb slices, 5 alpha-DHP represented 25.2 +/- 3.3% of total radioactivity and 3 alpha,5 alpha-THP 17.5 +/- 2.8%, whereas in rat medulla oblongata slices, 5 alpha-DHP was 31.3 +/- 3.5% and 3 alpha,5 alpha THP 5.4 +/- 1.5% of total radioactivity. In slices from other rat brain regions, both metabolites represented 12-20% of total radioactivity. The highest metabolite content in mouse brain was also detected in olfactory bulb slices, where 5 alpha-DHP represented 16.6 +/- 4.6% and 20 alpha-DHP 9.5 +/- 2.3% of total radioactivity. In cortical and corpus callosum slices of monkey brain, 26.8 +/- 4.4% and 2.4 +/- 0.5% of total radioactivity, respectively, were converted to 20 alpha-DHP, and less than 3% of total radioactivity could be attributed to any of the other metabolites detected. The 3 alpha, 5 alpha-THP content in both rat and monkey brain was below 1 nM, but increased in rat brain to 6.7 +/- 2.5 nM after electroshock. Endogenous 3 alpha,5 alpha-THP might play an important role in the regulation of rat behavior through the modulation of GABA action on the GABAA receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245960 TI - Expression of facilitative glucose transporter isoforms in human brain tumors. AB - The expression of facilitative glucose transporter (GLUT) isoforms in human astrocytic tumors was examined. Reverse transcriptase-polymerase chain reaction of a surgically biopsied glioblastoma was carried out using the degenerative oligonucleotide primers corresponding to the sequences of the human facilitative glucose transporter family, and polymerase chain reaction products were hybridized with human GLUT1, GLUT2, GLUT3, GLUT4, and GLUT5 cDNA probes. The results showed that a biopsied glioblastoma expressed GLUT1, GLUT3, and GLUT4 glucose transporter genes. Northern blot analysis of total RNA (10 micrograms) from a biopsied glioblastoma showed the transcripts of only GLUT1 and GLUT3, suggesting that the expression of insulin-responsive glucose transporter GLUT4 mRNA is relatively low. Immunoblot analysis of biopsied glioblastoma tissues by polyclonal antibodies against the C-terminal synthetic peptides of GLUT1, GLUT3, and GLUT4 showed a single band of each polypeptide. However, elevated expression of GLUT1 and GLUT3 glucose transporters was not observed in the glioblastoma. Astrocytic tumor tissues (n = 14) were also examined immunohistochemically. Reactive products for GLUT1 were observed in the luminal surface of capillaries in all cases, whereas tumor cells were positive for GLUT1 in only two of 14 cases. GLUT3 was positive in astrocytic tumor cells in all cases. Three of 14 cases expressed the GLUT4 protein, which was localized in the cytoplasm of tumor cells. These results suggest that the facilitative glucose transport may be altered in astrocytic tumor cells and thus display a significant change in glucose metabolism. PMID- 8245961 TI - Regulated expression of the neurofibromin type I transcript in the developing chicken brain. AB - Neurofibromatosis type 1 (NF-1) is among the most common inherited diseases affecting cells of the central and peripheral nervous systems. A region of the NF 1 gene is similar in sequence to the ras-GTPase activator protein (ras-GAP), and investigations have confirmed that the NF-1 gene product (now known as neurofibromin) stimulates ras-GTPase activity in vitro and in vivo. Neurofibromin modulates the ability of ras proteins to regulate cellular proliferation and/or differentiation, suggesting a possible role in normal development. An alternative form of the neurofibromin transcript with an additional 63-bp exon inserted in the GAP-related domain (GRD) has been described recently. To determine whether differential expression of the two forms of neurofibromin GRD mRNA plays a role in embryonic development, we have isolated and characterized the corresponding chicken cDNA. The predicted amino acid sequence for the inserted exon is identical between chick and human, as are the exon-intron boundaries. RNase protection and RNA-polymerase chain reaction analyses demonstrate that most tissues express predominantly type II mRNA (which contains the insert) throughout embryonic development. In contrast, whereas type II is the major form in the brain early in development, expression of the type I transcript (without the insert) in this tissue increases dramatically at later times. Analysis of primary cultures derived from chick embryo brain indicates that the type I mRNA is enriched in neurons. PMID- 8245962 TI - Kynurenine pathway enzymes in brain: responses to ischemic brain injury versus systemic immune activation. AB - Accumulation of L-kynurenine and quinolinic acid (QUIN) in the brain occurs after either ischemic brain injury or after systemic administration of pokeweed mitogen. Although conversion of L-[13C6]tryptophan to [13C6]-QUIN has not been demonstrated in brain either from normal gerbils or from gerbils given pokeweed mitogen, direct conversion in brain tissue does occur 4 days after transient cerebral ischemia. Increased activities of enzymes distal to indoleamine-2,3 dioxygenase may determine whether L-kynurenine is converted to QUIN. One day after 10 min of cerebral ischemia, the activities of kynureninase and 3-hydroxy 3,4-dioxygenase were increased in the hippocampus, but local QUIN levels and the activities of the indoleamine-2,3-dioxygenase and kynurenine-3-hydroxylase were unchanged. By days 2 and 4 after ischemia, however, the activities of all these enzymes in the hippocampus as well as QUIN levels were significantly increased. Kynurenine aminotransferase activity in the hippocampus was unchanged on days 1 and 2 after ischemia but was decreased on day 4, at a time when local kynurenic acid levels were unchanged. A putative precursor of QUIN, [13C6]anthranilic acid, was not converted to [13C6]QUIN in the hippocampus of either normal or 4-day post ischemic gerbils. Gerbil macrophages stimulated by endotoxin in vitro converted L [13C6]tryptophan to [13C6]QUIN. Kinetic analysis of kynurenine-3-hydroxylase activity in the cerebral cortex of postischemic gerbils showed that Vmax increased, without changes in Km. Systemic administration of pokeweed mitogen increased indoleamine-2,3-dioxygenase and kynureninase activities in the brain without significant changes in kynurenine-3-hydroxylase or 3-hydroxyanthranilate 3,4-dioxygenase activities. Increases in kynurenine-3-hydroxylase activity, in conjunction with induction of indoleamine-2,3-dioxygenase, kynureninase, and 3 hydroxyanthranilate-3,4-dioxygenase in macrophage infiltrates at the site of brain injury, may explain the ability of postischemic hippocampus to convert L [13C6]tryptophan to [13C6]QUIN. PMID- 8245963 TI - Developmental changes in tau phosphorylation: fetal tau is transiently phosphorylated in a manner similar to paired helical filament-tau characteristic of Alzheimer's disease. AB - Rat and human fetal brain tau were probed with a panel of monoclonal antibodies (tau-1, AT8, 8D8, RT97, SMI31, SMI34) that distinguish between paired helical filament (PHF)-tau of Alzheimer's disease and normal adult brain tau. These antibodies discriminate between normal and PHF-tau because their epitopes are phosphorylated in PHF-tau. Although only one molecular isoform of tau was shown to be expressed in fetal brain, two fetal tau species could be distinguished on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the slower migrating species was recognized by all of the PHF-tau-specific antibodies. Moreover, this immunoreactivity was shown to be phosphorylation dependent. Our observations suggest that the abnormal phosphorylation of tau in Alzheimer's disease may be the result of reactivation of pathways governing the phosphorylation of tau in the developing brain. PMID- 8245964 TI - Interconvertible kinetic states of t-butylbicycloorthobenzoate binding sites of the gamma-aminobutyric acidA ionophores. AB - The kinetics of t-[3H]butylbicycloorthobenzoate (TBOB) binding to the convulsant sites of the gamma-aminobutyric acidA (GABAA) receptor-ionophore complex were examined in synaptosomal membrane preparations of rat brain. On and off rates of TBOB binding were accelerated by 1 microM GABA and decelerated by 1 microM bicuculline methochloride, a GABAA antagonist. The presence of GABA and bicuculline methochloride created rapid and slow phases of dissociation, respectively. The three groups of rate constants distinguished for the dissociation of 4 nM and 30 nM [3H]TBOB represent multi-affinity states of the convulsant sites depending on the presence of GABA or bicuculline methochloride. Apparent association rate constants do not obey the equation k(app) = k(off) + k(on) [TBOB] without assuming interconvertibility of the kinetic states during binding. Avermectin B1a (AVM B1a), a chloride channel opening agent, accelerated the association and dissociation of TBOB and resulted in a biphasic effect on TBOB binding, i.e., enhancement at low concentrations (EC50 7.8 nM) followed by displacement at high concentrations (IC50 6.3 microM) of AVM B1a. AVM B1a resulted in similar biphasic effects on t-[35S]butylbicyclophosphorothionate binding. DIDS, an isothiocyanatostilbene derivative with irreversible anion channel blocking effect, selectively inhibited basal [3H]TBOB binding (IC50 125 microM DIDS) leaving the enhancement by AVM B1a unaffected. PMID- 8245965 TI - Alterations in actin-binding beta-thymosin expression accompany neuronal differentiation and migration in rat cerebellum. AB - The beta 4- and beta 10-thymosins, recently identified as actin monomer sequestering proteins, are developmentally regulated in brain. Using specific mRNA and protein probes, we have used in situ hybridization and immunohistochemical techniques to investigate the distribution of the beta thymosin mRNAs and their proteins in developing rat cerebellum. Early in postnatal development, both beta-thymosin mRNAs were expressed at highest levels in the postmitotic, premigratory granule cells of the external granular layer; expression diminished as granule cells migrated to and differentiated within the developing internal granular layer. In addition, both beta-thymosin proteins were present in bundles of cerebellar afferent fibers in the white matter at this time. Throughout the maturation period, both proteins were present in elongating parallel fibers in the upper portion of the molecular layer. Later in cerebellar development, thymosin beta 4, but not thymosin beta 10, was expressed in Golgi epithelial cells and Bergmann processes. Thymosin beta 4 was expressed in a small population of cells with microglial morphology scattered throughout the gray and white matter. Thymosin beta 10 was detected in an even smaller population of glia. Expression of thymosin beta 4 and thymosin beta 10 in premigratory granule cells and in growing neuronal processes is consistent with the possibility that both beta-thymosins are involved in the dynamics of actin polymerization during migration and process extension of neurons. PMID- 8245966 TI - pH-dependent binding of synthetic beta-amyloid peptides to glycosaminoglycans. AB - The senile plaques found within the cerebral cortex and hippocampus of the Alzheimer disease brain contain beta-amyloid peptide (A beta) fibrils that are associated with a variety of macromolecular species, including dermatan sulfate proteoglycan and heparan sulfate proteoglycan. The latter has been shown recently to bind tightly to both amyloid precursor protein and A beta, and this binding has been attributed largely to the interaction of the core protein of heparan sulfate proteoglycan with A beta and its precursor. Here we have examined the ability of synthetic A beta s to bind to and interact with the glycosaminoglycan moieties of proteoglycans. A beta(1-28) associates with heparin, heparan sulfate, dermatan sulfate, and chondroitin sulfate. The interaction of these sulfated polysaccharides with the amyloid peptide results in the formation of large aggregates that are readily sedimented by centrifugation. The ability of both A beta(1-28) and A beta(1-40) to bind glycosaminoglycans is pH-dependent, with increasing interaction as the pH values fall below neutrality and very little binding at pH 8.0. The pH profile of heparin-induced aggregation of A beta(1-28) has a midpoint pH of approximately 6.5, suggesting that one or more histidine residues must be protonated for binding to occur. Analysis of the A beta sequence reveals a consensus heparin-binding domain at residues 12-17, and this motif contains histidines at positions 13 and 14 that may be involved in the interaction with glycosaminoglycans.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245967 TI - Microglia-derived elastase produces a low-molecular-weight plasminogen that enhances neurite outgrowth in rat neocortical explant cultures. AB - In the course of analysis of plasminogen in microglial conditioned medium (Mic CM), novel low-molecular-weight (LMW) zymogen with a molecular mass of approximately 36 kDa was detected by casein-urokinase zymography. Because this form was produced when rat native plasminogen was incubated with Mic-CM, a specific protease in the Mic-CM was thought to be responsible for the production of LMW plasminogen. The production of LMW plasminogen was strongly inhibited by elastase inhibitors. Furthermore, elastase (pancreatic or leukocyte) was also found to produce LMW zymogen from native plasminogen. These results indicate that LMW plasminogen is produced through limited proteolysis by an elastase-like protease in Mic-CM. To determine the biochemical characteristics of LMW plasminogen, rat native plasminogen was cleaved by pancreatic elastase, and the fragments (LMW plasminogen and nonzymogen fragments) were purified by several kinds of column chromatography. Amino acid sequence analysis revealed that LMW plasminogen is a carboxy-terminal region that contains the fifth kringle domain and a protease active site, and the amino acid sequence is identical to that of LMW plasminogen produced by Mic-CM. On the other hand, the nonzymogen fragment was the amino-terminal region containing four kringle domains. The effects of native plasminogen and the fragments on neurite outgrowth of rat brain explant were examined. LMW plasminogen promoted neurite outgrowth as well as did native plasminogen, whereas nonzymogen fragments did not. These results suggest that LMW plasminogen, which is produced from native plasminogen by elastase, may be a physiologically active molecule that mediates the intercellular interaction between microglia and neurons. PMID- 8245968 TI - Soluble and particulate organophosphorus neuropathy target esterase in brain and sciatic nerve of the hen, cat, rat, and chick. AB - Considerable evidence exists suggesting that the so-called neuropathy target esterase (NTE) is involved in the mechanisms responsible for organophosphorus induced delayed polyneuropathy (OPIDP). Earlier studies in the adult hen, the habitually employed experimental model in OPIDP, have shown that most NTE activity in the brain is centered in particulate fractions, whereas approximately 50% of this activity in the sciatic nerve is encountered in soluble form, with the rest being particulate NTE. In the present work, we have studied the particulate and soluble fractional distribution of paraoxon-resistant phenylvalerate esterase activity (B activity), paraoxon- and mipafox-resistant phenylvalerate esterase activity (C activity), and NTE activity (B-C) according to ultracentrifugation criteria (100,000 g for 1 h). To this effect, two sensitive (adult hen and cat) and two scarcely sensitive (rat and chick) models were used. In all four experimental models, the distribution pattern was qualitatively similar: B activity and total NTE were much greater in brain (900 2,300 nmol/min/g of tissue) than in sciatic nerve (50-100 nmol/min/g of tissue). The proportion of soluble NTE in brain was very low (< 2%), whereas its presence in sciatic nerve was substantial (30-50%). The NTE/B ratio in brain was high for the particulate fraction (> 60%) and low in the soluble fraction (7-30%); in sciatic nerve the ratio was about 50% in both fractions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245969 TI - Involvement of adenosine-sensitive cyclic AMP-generating systems in cobalt induced epileptic activity in the rat. AB - An injection of cobalt chloride solution into the unilateral sensorimotor cortex of rats induced electrographic epileptic activity, which was followed by a peripheral motor disturbance. Brain slices were prepared from the cortical region including the injection site and from the other cortical regions of rats between 8 and 50 days after the injection. In the cortical slices, we examined cyclic AMP accumulations elicited by adenosine and its stable analogue 2-chloroadenosine. Adenosine and 2-chloroadenosine at their maximal dose increased cyclic AMP accumulation six- to 10-fold and 10-15-fold, respectively, and the elicitation was markedly inhibited by the adenosine antagonist 8-phenyltheophylline. The cyclic AMP accumulation was increased in the primary epileptic region of the cortex adjacent to the injection site of cobalt chloride solution, whereas it was unchanged in the other cortical regions. The increase in cyclic AMP accumulation was observed regardless of the presence or absence of the adenosine uptake inhibitor dipyridamole, the phosphodiesterase inhibitor DL-4-(3-butoxy-4 methoxybenzyl)-2-imidazolidinone, and adenosine deaminase. Such an increased accumulation of cyclic AMP in the primary epileptic cortex was detected as early as 8 days after the injection. The cyclic AMP accumulation continued to increase and reached a peak level 17-19 days after the injection, and it returned to the control levels after 40-50 days, in correspondence with the electrographic and behavioral findings. It is concluded that alterations in adenosine receptor mediated generation of cyclic AMP in the primary epileptic cortex are closely associated with the central process of cobalt-induced epilepsy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245970 TI - Extracellular glutamate is increased in thalamus during thiamine deficiency induced lesions and is blocked by MK-801. AB - The current study measured extracellular fluid (ECF) levels of excitatory amino acids before and during the onset of thiamine deficiency-induced pathologic lesions. Male Sprague-Dawley rats were treated with daily pyrithiamine (0.25 mg/kg i.p.) and a thiamine-deficient diet (PTD). Microdialysates were simultaneously collected from probes inserted acutely via guide cannulae into right paracentral and ventrolateral nuclei of thalamus and left hippocampus of PTD and pair-fed controls. Hourly samples were collected from unanesthetized and freely moving animals. Basal levels obtained at a prelesion stage (day 12 of PTD treatment) were unchanged from levels in pair-fed controls. In samples collected 4-5 h after onset of seizures (day 14 of PTD), the levels of glutamate were elevated an average 640% of basal levels in medial thalamus and 200% in hippocampus. Glutamine levels declined, taurine and glycine were elevated, and aspartate, GABA, and alanine were unchanged during this period. Within 7 h after seizure onset glutamine was undetectable in both areas, whereas glutamate had declined to approximately 200% in thalamus and 70% in hippocampus. No significant change in glutamate, aspartate, or other amino acids was observed in dialysates collected from probes located in undamaged dorsal-lateral regions of thalamus. Number of neurons within ventrolateral nucleus of thalamus was significantly greater in PTD animals in which the probe was dialyzed compared with nondialyzed, suggesting that removal of excitatory amino acids was protective. No significant pathologic damage was evident in hippocampus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245971 TI - Synergistic interaction of muscarinic and opioid receptors with GS-linked neurotransmitter receptors to stimulate adenylyl cyclase activity of rat olfactory bulb. AB - We reported previously that in homogenates of rat olfactory bulb muscarinic and opioid receptor agonists stimulate adenylyl cyclase activity. In the present study we show that carbachol (CCh) and Leu-Enkephalin act synergistically with vasoactive intestinal peptide (VIP) and corticotropin-releasing hormone (CRH), but not with l-isoproterenol, in increasing cyclic AMP formation. The synergistic interaction consists of an increase in the maximal adenylyl cyclase activation without a significant change in the potency of each agonist. CCh also fails to affect 125I-CRH binding to olfactory bulb membranes. The synergism requires micromolar concentrations of GTP. Substitution of the stable GTP analog guanosine 5'-O-(3'-thiotriphosphate) for GTP allows the CRH stimulation, but abolishes the CCh enhancement of both basal and CRH-stimulated enzyme activities. Moreover, in vivo treatment of olfactory bulbs with pertussis toxin completely prevents the muscarinic and opioid effects. Thus, the synergistic interaction appears to result from opioid- and muscarinic-induced activation of a pertussis toxin sensitive GTP-binding protein which may potentiate the adenylyl cyclase stimulation by the stimulatory GTP-binding protein activated by either VIP or CRH receptors. PMID- 8245972 TI - Neither moderate hypoxia nor mild hypoglycaemia alone causes any significant increase in cerebral [Ca2+]i: only a combination of the two insults has this effect. A 31P and 19F NMR study. AB - (1) The energy state and free intracellular calcium concentration ([Ca2+]i) of superfused cortical slices were measured in moderate hypoxia (approximately 65 microM O2), in mild hypoglycaemia (0.5 mM glucose), and in combinations of the two insults using 19F and 31P NMR spectroscopy. (2) Neither hypoxia nor hypoglycaemia alone caused any significant change in [Ca2+]i. Hypoxia caused a 40% fall in phosphocreatine (PCr) content but not in ATP level, and hypoglycaemia produced a slight fall in both (as expected from previous studies). These changes in the energy state recovered on return to control conditions. (3) A combined sequential insult (hypoxia, followed by hypoxia plus hypoglycaemia) produced a 100% increase in [Ca2+]i and a decrease in PCr level to approximately 25% of control. The reverse combined sequential insult (hypoglycaemia, followed by hypoglycaemia plus hypoxia) had the same effect. On return to control conditions there was some decrease in [Ca2+]i and a small increase in PCr content, but neither recovered to control levels. (4) Exposure of the tissue to the combined simultaneous insult (hypoxia plus hypoglycaemia) immediately after the control spectra had been recorded resulted in a fivefold increase in [Ca2+]i and a similar decrease in PCr level to 20-25% of control. There was little if any change of [Ca2+]i or PCr level on return to control conditions. (5) These results are discussed in terms of metabolic adaptation of some but not all of the cortical cells to the single type of insult, which renders the tissues less vulnerable to the combined insult. PMID- 8245973 TI - Decline in response to nicotine in aged rat striatum: correlation with a decrease in a subpopulation of nicotinic receptors. AB - Specific and reproducible changes involving the cholinergic and dopaminergic systems have been described in both the aging rodent and the human nervous system. Nevertheless, relatively little information is available on changes in nicotinic cholinergic receptors occurring in normal aging, and there have been few attempts to correlate alterations in receptor densities with changes in nicotinic actions. We have utilized the nicotine-mediated stimulation of endogenous dopamine efflux in a striatal slice preparation as a functional index of responsiveness to nicotine in aging. Following incubation with nicotine, this efflux was significantly lower in 25-month-old (aged) as opposed to 4-month-old (young) rats. In contrast, the release of striatal dopamine following a high potassium stimulus was similar at both ages. Binding studies in young and aged animals did not reveal any significant change with age in the total number of striatal nicotinic receptors recognized by either [3H]nicotine or the neuronal nicotinic antagonist 125I-neuronal bungarotoxin. However, there was a nearly 80% decline in the subpopulation of striatal nicotinic receptors jointly recognized by both nicotine and neuronal bungarotoxin, but not by alpha-bungarotoxin. Quantitative autoradiography demonstrated declines with age in this receptor subtype in several brain regions examined. Decrements in this specific subpopulation of nicotinic receptors or in the nerve cells expressing these receptors may contribute to the functional declines that take place in the aging motor and visual systems. PMID- 8245974 TI - Developmental regulation of mRNAs encoding rat brain kainate/AMPA receptors: a northern analysis study. AB - Functionally diverse kainate/alpha-amino-3-hydroxy-5-methyl-4- isoxazolepropionic acid (AMPA) receptors are generated by assembly of glutamate receptor (GluR)1, 2, and 3 subunits into homomeric and heteromeric channels. We examined GluR1, 2, and 3 gene expression in embryonic, neonatal, and adult rat brain by northern analysis under conditions of high stringency. In the adult, hybridization to a GluR1 riboprobe revealed the presence of an abundant RNA species, 5.2 kb in size, and minor bands of 3.2 and 3.9 kb. GluR2 hybridized to two species, 3.9 and 5.9 kb, of comparable abundance, presumably attributable to alternate splice products. Hybridization to the GluR3 riboprobe showed a major species of 5.2 kb. This pattern of RNA species was invariant over all the brain regions examined. Examination of GluR expression in development revealed that in the postnatal period, GluR1, 2, and 3 mRNAs are regulated as a function of age. In adult rat brain, GluR1 and 2 mRNA expression was highest in hippocampus; GluR3 was expressed at highest density in hippocampus and frontal cortex. The three transcripts were first detected at embryonic day 16 and then exhibited changes in expression levels in a region-specific manner. In hippocampus, all three transcripts exhibited elevated expression in the late neonatal period; in frontal cortex, elevated expression was observed for GluR2 and 3 only. In striatum, all three transcripts were expressed at relatively low levels throughout development, with a modest peak at postnatal day 14. In cerebellum, the GluR1 mRNA level was high from postnatal day 28 to adult.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8245975 TI - The ontogeny of apomorphine-induced alterations of neostriatal dopamine release: effects on spontaneous release. AB - The effects of apomorphine (0.05, 0.1, and 1.0 mg/kg, s.c.) on the extracellular levels of dopamine and the dopamine metabolite 3,4-dihydroxyphenylacetic acid were studied through the use of in vivo microdialysis in the neostriatum of developing and adult rats. Fifteen-minute samples were collected from urethane anesthetized rats 5, 10-11, 21-22, and 35-36 days old and adults and quantified by HPLC with electrochemical detection. Apomorphine attenuated extracellular levels of dopamine in all age groups, suggesting that the dopamine autoreceptor modulating release in the neostriatum is functional by 5 days of age. A dose response effect of apomorphine on extracellular dopamine was observed in all age groups except at 10-11 days of age. Extracellular levels of 3,4 dihydroxyphenylacetic acid were also significantly decreased in all age groups, consistent with the hypothesis that synthesis-modulating dopamine autoreceptors in the neostriatum are functional by 5 days of age. Apomorphine had a significantly greater effect on extracellular 3,4-dihydroxyphenylacetic acid levels at the 0.05 and 0.1 mg/kg doses in the 5- and 10-11-day-old age groups compared with the other ages. Absolute levels of extracellular dopamine were significantly attenuated at 5 days of age compared with the other ages, and absolute levels of extracellular 3,4-dihydroxyphenylacetic acid monotonically increased with age. PMID- 8245976 TI - ucb 11056, a new potential nootropic drug, amplifies induced cyclic AMP formation in rat brain tissue. AB - ucb 11056 [2-(4-morpholino-6-propyl-1,3,5-triazin-2-yl)aminoethanol] induced a significant (approximately 25%) increase in cyclic AMP levels in different brain areas following its intraperitoneal injection. This effect started as early as 2 min postinjection and lasted for 30 min, after which cyclic AMP levels returned to normal. In hippocampal slice preparations in vitro, ucb 11056 exerted a strong potentiation of cyclic AMP levels when it was combined with agents such as norepinephrine, forskolin, and isoproterenol. Only a slight effect on cyclic AMP levels was measured when ucb 11056 was incubated alone with hippocampal slices. The potentiating effect of ucb 11056 on norepinephrine-stimulated cyclic AMP formation was partially reduced when slices were pretreated with yohimbine and totally abolished when slices were treated with propranolol. These combined data indicate that (a) ucb 11056 rapidly increases cyclic AMP levels in the rat brain in vivo and (b) ucb 11056 potentiates stimulated cyclic AMP formation in vitro. The data also suggest that the central effect of ucb 11056 might be via the modulation of cyclic AMP generation, most probably mediated through adenylate cyclase activation mechanisms combined with a weak inhibitory activity on the cyclic nucleotide phosphodiesterase activity. PMID- 8245977 TI - Clearance of exogenous dopamine in rat dorsal striatum and nucleus accumbens: role of metabolism and effects of locally applied uptake inhibitors. AB - In vivo electrochemistry was used to investigate the mechanisms contributing to the clearance of locally applied dopamine in the dorsal striatum and nucleus accumbens of urethane-anesthetized rats. Chronoamperometric recordings were continuously made at 5 Hz using Nafion-coated carbon fiber electrodes. When a finite amount of dopamine was pressure-ejected at 5-min intervals from a micropipette adjacent to the electrode, transient and reproducible dopamine signals were detected. Substitution of L-alpha-methyldopamine, a substrate for the dopamine transporter but not for monoamine oxidase, for dopamine in the micropipette did not substantially alter the time course of the resulting signals. This indicates that metabolism of locally applied dopamine to 3,4 dihydroxy-phenylacetic acid is not responsible for the decline in the dopamine signal. Similarly, changing the applied oxidation potential from +0.45 to +0.80 V, which allows for detection of 3-methoxytyramine formed from dopamine via catechol-O-methyltransferase, had little effect on signal amplitude or time course. In contrast, lesioning the dopamine terminals with 6-hydroxydopamine, or locally applying the dopamine uptake inhibitors cocaine or nomifensine before pressure ejection of dopamine, significantly increased the amplitude and time course of the dopamine signals in both regions. The effects of cocaine and nomifensine were greater in the nucleus accumbens than in the dorsal striatum. Local application of lidocaine and procaine had no effect on the dopamine signals. Initial attempts at modeling resulted in curves that were in qualitative agreement with our experimental findings. Taken together, these data indicate that (1) uptake of dopamine by the neuronal dopamine transporter, rather than metabolism or diffusion, is the major mechanism for clearing locally applied dopamine from the extracellular milieu of the dorsal striatum and nucleus accumbens, and (2) the nucleus accumbens is more sensitive to the effects of inhibitors of dopamine uptake than is the dorsal striatum. PMID- 8245978 TI - Ceramide 1-phosphate phosphatase activity in brain. AB - Recent studies have implicated sphingolipids in a variety of intracellular signaling systems. The finding that a calcium-stimulated ceramide kinase copurifies with neurotransmitter-containing vesicles suggests that ceramide, or one of its metabolites, has a role in neurotransmitter release. As a step toward understanding the role of ceramide kinase in vesicle functioning, this study sought to determine the metabolic fate of the product, ceramide 1-phosphate. We report that ceramide 1-phosphate is not deacylated by brain ceramidases to produce sphingosine 1-phosphate. It is, however, the substrate for a phosphatase activity that we name ceramide 1-phosphate phosphatase (CPPase). Subcellular fractionation studies suggest that CPPase is found in the synaptic terminal and is associated with both synaptic vesicle and plasma membranes. Divalent cations, most notably calcium, inhibit CPPase activity although not at concentrations that activate ceramide kinase. The existence of both ceramide kinase and CPPase activities at the synapse suggests that ceramide 1-phosphate production regulates some aspect of synaptic vesicle functioning. PMID- 8245979 TI - Sex differences in acute swim stress-induced changes in the binding of MK-801 to the NMDA subclass of glutamate receptors in mouse forebrain. AB - Acute swim stress (3 min at 32 degrees C) in mice produces increases in the binding of MK-801 to the NMDA subclass of glutamate receptors to forebrain membranes prepared from male mice. Scatchard analyses indicate that the observed increases in the binding of MK-801 in membranes from male mice are the result of changes in the affinity and density of low-affinity binding sites and in the density of high-affinity binding sites. In female mice, any changes in the binding of MK-801 appear to be much less pronounced and restricted to the low affinity binding sites. These results are in contrast to the situation with binding to GABA receptors where acute swim stress increases GABA binding in forebrain membranes much more in female than in male mice. This indicates significant sex differences in the responses of receptors for the major excitatory and inhibitory transmitters to acute swim stress. These rapid changes in MK-801 binding may result from changes in endogenous modulators as appears to be the case in the acute swim stress-induced changes in GABA binding. As with GABA binding, the endogenous modulators are likely to include steroids, the sex differences reflecting differences in modulation by gonadal steroids and the stress-induced changes reflecting differences in modulation by adrenal steroids. Estradiol, progesterone, and corticosterone treatments have been reported by other workers to influence the properties of glutamate receptors. PMID- 8245980 TI - Sequence and novel distribution of the chicken homologue of the mammalian gamma aminobutyric acidA receptor gamma 1 subunit. AB - cDNAs have been cloned that encode the chicken (Gallus domesticus) gamma aminobutyric acidA receptor gamma 1 subunit, the mature sequence of which shares 90, 79, and 69% identity with those of the rat gamma 1, gamma 2, and gamma 3 subunits, respectively. In situ hybridization reveals that there are pronounced differences in the regional and cellular localizations of the corresponding gamma aminobutyric acidA receptor gamma-subunit mRNA compared with that of the gamma 2 subunit mRNA in 1-day-old chick brain. The absence of the gamma 1-subunit transcript in certain chick brain nuclei of visual and auditory pathways, in which gamma 2-subunit mRNA is present, points to differences in the functional roles of receptors containing one or other of these polypeptides. Certain cells in other brain regions appear to contain both gamma 1- and gamma 2-subunit mRNAs, suggesting that they either have two gamma-aminobutyric acidA receptor subtypes or possess receptors incorporating two different gamma subunits. We have also found contrasts in the distribution patterns, in homologous brain regions, of the chicken gamma 1-subunit mRNA and the rat gamma 1-subunit mRNA. These data may reflect different functional roles of the chicken and rat gamma 1 subunits. PMID- 8245981 TI - Lithium decreases membrane-associated protein kinase C in hippocampus: selectivity for the alpha isozyme. AB - We investigated the effects of lithium on alterations in the amount and distribution of protein kinase C (PKC) in discrete areas of rat brain by using [3H]phorbol 12,13-dibutyrate quantitative autoradiography as well as western blotting. Chronic administration of lithium resulted in a significant decrease in membrane-associated PKC in several hippocampal structures, most notably the subiculum and the CA1 region. In contrast, only modest changes in [3H]phorbol 12,13-dibutyrate binding were observed in the various other cortical and subcortical structures examined. Immunoblotting using monoclonal anti-PKC antibodies revealed an isozyme-specific 30% decrease in hippocampal membrane associated PKC alpha, in the absence of any changes in the labeling of either the beta (I/II) or gamma isozymes. These changes were observed only after chronic (4 week) treatment with lithium, and not after acute (5 days) treatment, suggesting potential clinical relevance. Given the critical role of PKC in regulating neuronal signal transduction, lithium's effects on PKC in the limbic system represent an attractive molecular mechanism for its efficacy in treating both poles of manic-depressive illness. In addition, the decreased hippocampal membrane-associated PKC observed in the present study offers a possible explanation for lithium-induced memory impairment. PMID- 8245982 TI - Determination of kainic acid-induced release of nitric oxide using a novel hemoglobin trapping technique with microdialysis. AB - We have demonstrated the usefulness of a novel hemoglobin-trapping technique to quantify nitric oxide (NO) concentrations in vivo. Concentric microdialysis probes were implanted into the hippocampus of rats under urethane anesthesia and perfused with 1 microM oxyhemoglobin in artificial CSF to sequester NO in extracellular fluid. The concentration of methemoglobin was then determined spectrophotometrically. The basal level of NO in hippocampus was 2.2 +/- 0.5 nM (in vitro sensitivity of the probe was 0.2 nM). Administration of 13 mg/kg, i.p., of kainic acid (KA) produced a maximal 5.3-fold increase at 100 min in NO levels (11.8 +/- 0.2 nM). This response was significantly attenuated by pretreatment with the NO synthase inhibitor N-monomethyl-L-arginine (50 mg/kg, 30 min before KA). These results demonstrate that a microdialysis probe using a novel hemoglobin-trapping technique possesses adequate sensitivity to determine the basal levels of NO and document the ability of KA to increase these levels via a NO synthase-mediated mechanism. PMID- 8245983 TI - Functional identification and molecular cloning of a human brain vesicle monoamine transporter. AB - A vesicle monoamine transporter was functionally identified, molecularly cloned, and characterized from a human substantia nigra cDNA library. The ATP-dependent transport of 5-[3H]hydroxytryptamine ([3H]5-HT) by digitonin-permeabilized fibroblasts expressing the vesicle monoamine/H+ antiporter in culture exhibited a Km of 0.55 microM. Reserpine and tetrabenazine, inhibitors of two monoamine binding sites, effectively blocked [3H]5-HT accumulation with Ki values of 34 and 78 nM, respectively. Pretreatment of cells with as little as 10 nM reserpine in the presence of ATP abolished uptake. The rank order for substrate inhibition of [3H]5-HT uptake for both the previously reported rat vMAT1 and the human transporter clone followed the order 5-HT > dopamine > epinephrine > norepinephrine > 1-methyl-4-phenylpyridinium > 2-phenylethylamine > histamine. The virtually identical transport characteristics of rvMAT1 and hvMAT1 confirm the relevance of neuropharmacological studies of rat brain biogenic amine uptake and storage to human brain neurochemistry. PMID- 8245984 TI - Expression of the baculovirus p35 gene inhibits mammalian neural cell death. AB - Expression of the apoptosis suppressor gene p35, derived from the baculovirus Autographa californica nuclear polyhedrosis virus, markedly inhibited the cell death of stably transfected mammalian neural cells whether the cell death was induced by glucose withdrawal, calcium ionophore, or serum withdrawal. The p35 protein, which is required to block virus-induced apoptosis of cultured insect cells, is only the second gene product shown to block mammalian neural cell death, with Bcl-2 being the first. Because there is no apparent homology between p35 and Bcl-2, the existence of a cellular death program that may be modulated at multiple points is suggested. Furthermore, these findings demonstrate that the putative cellular death program is conserved across species and cell types. PMID- 8245985 TI - Superoxide dismutase activity, oxidative damage, and mitochondrial energy metabolism in familial and sporadic amyotrophic lateral sclerosis. AB - The cause of neuronal death in amyotrophic lateral sclerosis (ALS) is unknown. Recently, it was found that some patients with autosomal-dominant familial ALS (FALS) have point mutations in the gene that encodes Cu/Zn superoxide dismutase (SOD1). In this study of postmortem brain tissue, we examined SOD activity and quantified protein carbonyl groups, a marker of oxidative damage, in samples of frontal cortex (Brodmann area 6) from 10 control patients, three FALS patients with known SOD1 mutations (FALS-1), one autosomal-dominant FALS patient with no identifiable SOD1 mutations (FALS-O), and 11 sporadic ALS (SALS) patients. Also, we determined the activities of components of the electron transport chain (complexes I, II-III, and IV) in these samples. The cytosolic SOD activity, which is primarily SOD1 activity, was reduced by 38.8% (p < 0.05) in the FALS-1 patients and not significantly altered in the SALS patients or the FALS-O patient relative to the control patients. The mitochondrial SOD activity, which is primarily SOD2 activity, was not significantly altered in the FALS-1, FALS-O, or SALS patients. The protein carbonyl content was elevated by 84.8% (p < 0.01) in the SALS patients relative to the control patients. Finally, the complex I activity was increased by 55.3% (p < 0.001) in the FALS-1 patients relative to the control patients. These results from cortical tissue demonstrate that SOD1 activity is reduced and complex I activity is increased in FALS-1 patients and that oxidative damage to proteins is increased in SALS patients. PMID- 8245986 TI - Inhibition of beta-amyloid production by activation of protein kinase C. AB - The cellular factors regulating the generation of beta-amyloid from the amyloid precursor protein (APP) are unknown. Activation of protein kinase C (PKC) by phorbol ester treatment inhibited the generation of the 4-kDa beta-amyloid peptide in transfected COS cells, a human glioma cell line, and human cortical astrocytes. An analogue of diacylglycerol, the endogenous cellular activator of PKC, also inhibited the generation of beta-amyloid. Activation of PKC increased the level of secreted APP in transfected COS cells but did not significantly affect the level of secreted APP in primary human astrocytes or in the glioma cell line. Cell-associated APP and the secreted APP derivative, but not beta amyloid, were phosphorylated on serine residues. Activation of PKC did not increase the level of APP phosphorylation, suggesting that PKC modulates the proteolytic cleavage of APP indirectly by phosphorylation of other substrates. These results indicate that PKC activation inhibits beta-amyloid production by altering APP processing and suggest that beta-amyloid production can be regulated by the phospholipase C-diacylglycerol signal transduction pathway. PMID- 8245987 TI - Neurotoxicity of human amylin in rat primary hippocampal cultures: similarity to Alzheimer's disease amyloid-beta neurotoxicity. AB - Amylin, a 37-amino-acid amyloidogenic peptide, bears biophysical similarities to the amyloid-beta peptide (A beta) deposited in Alzheimer's disease. Using embryonic rat hippocampal cultures we tested whether amylin induces neurotoxicity similar to that previously observed with A beta(1-40). Treatment with human amylin(1-37) resulted in prominent toxicity as assessed by phase-contrast microscopy and quantification of lactate dehydrogenase in the medium. Amylin induced neurotoxicity was morphologically similar to that induced by A beta(1 40). In contrast, the nonamyloidogenic rat amylin showed negligible neurotoxicity despite having 95% sequence similarity to human amylin. Only full-length human amylin was toxic; various amylin peptide fragments including amino acid residues 20-29 were nontoxic at similar concentrations. These studies suggest that unrelated amyloidogenic peptides like human amylin and A beta can adopt a similar neurotoxic conformation in vitro. Similar conformation-dependent neurotoxicity may drive the prominent neurite degeneration around compacted but not diffuse deposits of A beta in Alzheimer's disease. PMID- 8245988 TI - GABA alters GABAA receptor mRNAs and increases ligand binding. AB - In addition to its role as an inhibitory neurotransmitter, gamma-aminobutyric acid (GABA) influences the cytodifferentiation of developing neurons both in culture and in vivo. Here, we report some of the targets of GABA action and the mechanism through which GABA acts. In primary cultures of cerebellar granule cells, GABA specifically stimulates an increase in the levels of mRNAs for alpha 1 and beta 2 GABAA receptor subunits. The GABAA agonist 4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP) mimics this effect, and the GABAA antagonist bicuculline prevents it. In addition, GABA and THIP trigger an increase in the number of GABA binding sites. This increase parallels that seen in vivo, where the total number of GABAA receptor sites increases during postnatal cerebellar development. It is interesting that the period of the greatest increase in the number of receptor sites coincides with the development of the granule cells. Taken together, our data suggest that GABA may play an important role during maturation of cerebellar granule cells by influencing the number and composition of its own receptors. PMID- 8245989 TI - Botulinum A like type B and tetanus toxins fulfils criteria for being a zinc dependent protease. AB - Although botulinum neurotoxin (BoNT) types A and B and tetanus toxin (TeTx) are specific inhibitors of transmitter release whose light chains contain a zinc binding motif characteristic of metalloendoproteases, only the latter two proteolyse synaptobrevin. Chelation of zinc or its readdition at high concentration hindered blockade of neuromuscular transmission by BoNT/A and B, indicating that type A also acts via a zinc-dependent mechanism. Such treatments prevented proteolysis of synaptobrevin II in rat brain synaptic vesicles by BoNT/B and TeTx but only the activity of the latter was antagonised appreciably by ASQFETS, a peptide spanning their cleavage site. The toxin's neuroparalytic activities were attenuated by phosphoramidon or captopril, inhibitors of certain zinc requiring proteases. However, these agents were ineffective in reducing the toxins' degradation of synaptobrevin except that a high concentration of captopril partially blocked the activity of TeTx but not BoNT/B, as also found for these drugs when tested on synaptosomal noradrenaline release. These various criteria establish that a zinc-dependent protease activity underlies the neurotoxicity of BoNT/A, a finding confirmed at motor nerve endings for type B and TeTx. Moreover, the low potencies of captopril and phosphoramidon in counteracting the toxins' effects necessitate the design of improved inhibitors for possible use in the clinical treatment of tetanus or botulism. PMID- 8245990 TI - Embolic complications of calf thrombosis following total hip arthroplasty. AB - The incidence of clinically significant pulmonary embolism following total hip arthroplasty is reported in patients with venographically documented deep venous thrombosis. One hundred seventy-four patients who enrolled in a prospective evaluation of deep venous thrombosis prophylaxis underwent contrast venography 7 10 days after operation. Ten patients with proximal and 12 patients with isolated deep calf thrombosis were identified on routine venogram reading and received appropriate anticoagulant therapy, including a 6-12-week course of warfarin. There were no clinically evident embolic events and no bleeding complications in this group. A secondary blinded review of venograms several weeks later identified isolated deep calf thrombi in 13 other patients, none of whom had received warfarin after discharge. Four of the 13 patients (P < .02) subsequently presented with pulmonary embolism at a mean of 33.5 days after operation. Asymptomatic deep calf venous thrombosis following total hip arthroplasty is associated with a significant risk of developing clinically evident pulmonary embolism within the first 8 weeks after operation. Once identified, these thrombi were effectively managed with outpatient warfarin anticoagulation, and both embolic and bleeding complications were avoided. This study documents the need for more prolonged routine prophylaxis against venous thrombosis than has heretofore been the rule, extending beyond the time of discharge from the hospital. Alternatively, the authors' experience supports the use of routine screening venography following total hip arthroplasty to allow detection and selective anticoagulant treatment of deep venous thrombosis in both the thigh and calf. PMID- 8245991 TI - Low-friction arthroplasty in severe acetabular dysplasia. AB - Eighty consecutive low-friction arthroplasties in 64 patients with secondary arthritis due to severe acetabular dysplasia (Crowe grades III and IV) are analyzed. Two groups of patients were considered. In group 1 over 20% of the cup was not covered by the acetabulum and was reconstructed with a femoral head graft (32 cases; in group 2) a femoral head graft was not used because the cup was adequately covered (48 cases). The mean follow-up period was 9.4 years (range, 3 17 years) for group 1 and 8.8 years (range, 3-18 years) for group 2. Early complications were frequent (19%). To study long-term results only 78 hips were assessed since two patients were excluded due to deep infection. In these 78 hips, group 1 patients showed "better" results after 16 years (69.6% good results) than group 2 (nongraft procedures) (52.2%), but these differences were not significant (P > or = .05). Early cup loosening was more frequent in group 2 than in group 1, and good coverage was obtained in 91% of the group 1 cups and in 71% of the group 2 cups (P > or = .05). Acetabular cup loosening was not correlated with acetabular cup position. Bone-grafts appeared consolidated in all cases, but resorption areas were frequently observed. Radiographic stem aseptic loosening appeared in 10 cases, cortical hypertrophy in 7 cases, and endosteal osteolysis in 3 cases. Although limb discrepancy was frequent in both groups (75%), the average discrepancy was 1.7 cm. PMID- 8245992 TI - Risk factors for recurrent aseptic loosening of the femoral component after cemented revision. AB - A series of 160 consecutive cemented first revisions, performed from 1977 through 1988 for aseptic loosening of a primary cemented femoral component, were studied using survivorship methods with the purpose of identifying risk factors for recurrent loosening of the femoral component. Risk of recurrent loosening depended on the length of the revision stem, with a significantly increased risk of loosening if the tip of the primary stem was overbridged by the revision stem with less than one width of the femoral shaft. Risk of recurrent loosening was also related to the extension of a cement mantle, exceeding more than 2 mm, around the revision stem measured on postrevision anteroposterior radiographs. Furthermore, low age and neutral position of the revision stem were identified as risk factors for recurrent loosening of the cemented revision femoral component. An improved fixation of the cemented revision femoral component for revisions performed after 1982 could be related to the use of longer revision stems and improved cementation. PMID- 8245993 TI - High assembly strains and femoral fractures produced during insertion of uncemented femoral components. A cadaver study. AB - The assembly strains produced in cadaver femurs during uncemented femoral arthroplasty were measured using strain gages and photoelastic coatings. Resecting the femoral neck, reaming the canal with power drills, and rasping with an optimal size rasp, as determined by preoperative radiographic templating, produced small assembly strains, up to 300 microstrain. Insertion of an optimal size prosthesis after preparing the femoral canal with instruments the same size as the prosthesis produced moderate assembly strains, up to 1,000 microstrain. Half a millimeter press-fit of optimal prostheses produced larger assembly strains, up to 2,000 microstrain. Half a millimeter press-fit of a prosthesis that was also one size (1.0 mm) larger than that determined to be optimum produced even larger assembly strains (2,000-6,000 microstrain) and longitudinal linear fractures in the femoral cortex. Insertion of prostheses that were smaller than the rasps produced minimal strains in the femoral cortex. The magnitude of peak strains produced by press-fitting the femoral components and the small amounts of disparity between the size of the recess and the prosthesis necessary to produce these strains show the narrow range of tolerances available to the surgeon. Cementless femoral arthroplasty requires great care in preparing the femoral canal to the appropriate size as determined from preoperative templating, using accurate and precisely toleranced instrumentation and prosthetic components in order to avoid femoral fractures, yet obtain a stable fit. PMID- 8245994 TI - Total knee arthroplasty in patellectomized patients. AB - Thirty-two total knee arthroplasties (TKAs) in patellectomized patients were evaluated with recent clinical and radiographic examinations. Eighteen patients had a primary TKA with a mean follow-up period of 49 months, and 14 patients had a revision TKA with a mean follow-up period of 36 months. A control group of 13 TKA patients with intact patellae were randomly generated but matched for age, sex, follow-up data, diagnosis, and prosthesis. Posterior cruciate ligament retaining types of prostheses were used in the primary knees, while the revision knees underwent arthroplasties with the more constrained, posterior cruciate ligament-substituting prostheses. All knees were evaluated based on the Knee Society's clinical and radiographic scoring system. In addition, 18 patients (9 primary, 9 revision) underwent isokinetic dynamometer testing for quantitation of peak quadriceps and hamstring torque. The knee score, indicative of pain relief, averaged 82.5 in the primary group (16 good/excellent, 1 fair, 1 poor), 86.5 in the revision group (12 good/excellent, 1 fair, 1 poor), and 93.9 in the control group (13/13 excellent). The function score averaged 59.7 in the primary group (6 good/excellent, 6 fair, 6 poor), 60.0 in the revision group (5 good/excellent, 2 fair, 7 poor), and 80.9 in the control group (12 good/excellent, 1 fair). The lower function scores predominantly reflected the patients' difficulty in independently climbing or descending stairs. This was also reflected in their higher flexion to extension peak torque ratios. There was one failure in the primary group requiring a revision and one failure in the revision group requiring a knee fusion. There was no radiographic evidence of impending failure in any of the remaining knees. No clinical or radiographic differences were found between the patients with osteoarthritis or rheumatoid arthritis. Although the knee and function scores were lower in the patellectomized patients, the overall results were generally satisfactory without a high incidence of failures. Satisfactory results were obtained in the primary TKAs using the minimally constrained prostheses when the posterior cruciate ligament was intact. Revision TKAs, in which the posterior cruciate ligament was absent, also demonstrated satisfactory results with the more constrained, posterior cruciate ligament substituting prostheses. PMID- 8245995 TI - Cutting errors in preparation of femoral components in total knee arthroplasty. AB - Cutting errors that occur during preparation of the femoral component can deter fixation in cementless total knee arthroplasty. This study evaluated the causes of femoral cutting errors and identified methods to minimize these errors. The cutting error caused by toggle of the saw blade in the guide-slot was measured with wood blocks, a cutting guide with 1.5 mm guide slots, and four kinds of saw blades (narrow and wide blades with thicknesses of standard 1.2 mm and thicker 1.4 mm). Another cause of cutting errors, relative motions between the cutting guide and distal femur due to vibration of the saw blade during bone cuts, was measured with preserved cadaver femurs. Various combined fixation methods with intramedullary rod, pins, and clamps were compared. The maximum cutting error caused by toggle of the saw blade in the slot was significantly reduced by using the thicker 1.4 mm saw blade, but the differences between wide and narrow blades in each thickness were not significant. Mean maximum cutting error at the cutting depth of 5 cm with 1.2 mm thick saw blades was 802 microns, and with 1.4 mm thick saw blades it was 488 microns. In the measurement of relative motion, fixation with only pins or clamps did not provide tight fixation and had deflections from 200 microns to more than 1 mm. The best fixation was obtained by combining fixation methods of intramedullary rod and pins or clamps. This resulted in total movement of less than 100 microns. The results of this study indicate that both toggle of the saw blade in the slot and motion of the cutting guide can cause major cutting errors. Cutting error can be minimized by using thicker saw blades and by fixing the cutting guide to the distal femur with combined fixation methods of a central rod and peripheral pins or clamps. PMID- 8245996 TI - Comparison of the subvastus and paramedian surgical approaches in bilateral knee arthroplasty. AB - A prospective randomized study was performed on 20 patients undergoing one-stage bilateral knee arthroplasty. One knee was exposed using a standard median parapatellar arthrotomy and the other knee with a subvastus arthrotomy. All patients underwent quantitative strength testing before surgery and at 1 week, 1 month, and 3 months after surgery. The knees were also evaluated for range of motion, and patients, who were blinded as to the approach used, completed questionnaires at each evaluation period as to their preference, if any, regarding knee pain and level of function. There was no difference in the range of motion between knees exposed with the paramedian or subvastus arthrotomy at any time period. The subvastus knees demonstrated significantly greater strength at the 1-week and 1-month intervals, but there was no strength difference at the 3-month interval. There were more lateral releases performed in the paramedium knees, and three minor complications were related to the subvastus approach. Patients who expressed a preference chose the subvastus knee 4:1 over the paramedian knee. The subvastus approach offers a reasonable alternative to the paramedian arthrotomy and preserves greater quadriceps strength in the early postoperative period. PMID- 8245997 TI - Revision total hip arthroplasty using ultrasonically driven tools. A clinical evaluation. AB - A three-center clinical evaluation of the use of ultrasonic instrumentation for cement removal during 90 revision total hip arthroplasties was undertaken. In conjunction with hand tools, cement removal was facilitated with ultrasonic tools. Superficial bone burns were seen in eight cases (9%). Bone perforations were created with ultrasonic tools in three cases (4%)--two followed perforations already created by hand tools in the proximal metaphyseal area. Only one perforation (1%) was made in the shaft of the femur. Ultrasonic instrumentation is a valuable adjunct for cement removal in revision total hip arthroplasty. PMID- 8245998 TI - Effect of postoperative reinfusion systems on hemoglobin levels in primary total hip and total knee arthroplasties. A prospective randomized study. AB - A prospective randomized study was undertaken to quantify the effect of reinfusion of postoperative shed blood drainage on the hemoglobin levels in patients undergoing elective primary total hip arthroplasty (THA) and total knee arthroplasty (TKA). One hundred eleven patients were enrolled between December 1990 and August 1991. There were 42 THAs and 69 TKAs. The study group consisted of 57 patients (35 TKAs and 22 THAs) who received a CBC ConstaVac (Stryker Surgical, Kalamazoo, MI) reinfusion system. The control group consisted of 54 patients (34 TKAs and 20 THAs) who received a ConstaVac collection unit. Postoperative drainage volumes were recorded for both groups. In addition, the volume of reinfused blood was recorded for the study group. Postoperative hemoglobins were recorded on postoperative days 1, 3, and 6, the latter reflecting the discharge hemoglobin level. All patients were encouraged to predeposit two units of autologous blood for both THAs and TKAs. This study showed no statistically significant difference in the postoperative hemoglobin levels between the study and control groups at anytime. Additionally, there was no statistically significant difference between hemoglobin levels and drainage volumes in both the THA and TKA study groups, compared to their respective control groups. The results of this study suggest that reinfusion units are not necessary in THAs and TKAs as a matter of routine use. PMID- 8245999 TI - Increased varus-valgus tilting of screw-fixated knee prostheses. Stereoradiographic study of uncemented versus cemented tibial components. AB - Twenty-six women and three men (34 knees) with osteoarthrosis were operated with the Miller-Galante I (Zimmer, Warsaw, IN) knee prosthesis. The patients were randomized to either cemented or uncemented fixation of the tibial component. All patients received a TiVaAl alloy tibial plate with four pegs and titanium fiber mesh undersurface. In the uncemented knees four screws were added. The fixation of the tibial component was determined by roentgen stereophotogrammetric analysis during the first 2 postoperative years. Rotations of the entire tibial component were recorded, as well as proximal or distal translation of various parts of the prosthetic edge corresponding to subsidence and lift-off. The uncemented components displayed almost all rotation and translation during the first 6 weeks, whereas the cemented ones displayed a more gradually increasing migration during the 2 years. Tibial component rotation about the sagittal axis was significantly increased in the uncemented knees throughout the investigation period. This corresponded to increased subsidence medially or laterally in the uncemented knees, whereas lift-off was equal in the two groups. Thin (8.5 mm) uncemented tibial components displayed more subsidence than the thicker ones at the medial or lateral edge 3 months after surgery. PMID- 8246000 TI - Rapidly progressive femoral head osteolysis. AB - Rapidly progressive osteolysis of the femoral head is an infrequently encountered yet rapidly disabling arthritic condition. This is a case of simultaneous osteolysis of the femoral heads in an otherwise healthy woman. Her clinical and laboratory findings support the diagnosis of rapidly progressive osteoarthritis, possibly accelerated by the use of indomethacin. Conventional total hip arthroplasty has successfully restored her ability to ambulate without pain, with stable implant fixation at the 2-year follow-up evaluation. PMID- 8246001 TI - Functional osseointegration of hydroxyapatite-coated implants in a weight-bearing canine model. AB - That hydroxyapatite (HA) can form a strong chemical bond with bone has been confirmed by several precedent workers using unloaded implants. The relationship between the histomorphometric measurements of the interface of the cementless femoral stems with and without HA coating and their mechanical properties in a weight-bearing canine model was evaluated in this study. Seven HA-coated and seven uncoated titanium (Ti6Al4V) hemiarthroplasties were implanted in the right hip of 14 dogs for a 5-month period. The specimens were taken to conduct mechanical testing and morphometric measuring. The average interface shear strength of 2.13 MPa for the HA-coated specimens was significantly higher than 0.93 MPa for the uncoated specimens (P < .05). There was greater bone apposition with the HA coating (72.39%) than without the coating (15.96%). Bone interfaces were divided into two types histomorphometrically: type A, simple bony shell without supporting trabeculae and type B, buttressed bony shell with supporting trabeculae. A positive correlation between the type B HA-bone interface and interface shear strength was found (r = .81). PMID- 8246002 TI - Variation of cortical hypertrophy depending on the medullary filling material. An experimental study of canine tibial diaphysis. AB - In order to evaluate the mineralization of bone after polymethyl methacrylate cementation, measurements of the periosteal apposition were made histologically and measurements of the bone-mineral content were made by dual-photon absorptiometry of the tibial diaphysis in 15 adult mongrel dogs in paired studies. Six dogs were investigated 4 weeks after intramedullary reaming of both sides and obturation of the medullary cavity with bone-wax on one side. Nine dogs were reamed on both sides. The medullary cavities were filled with polymethyl methacrylate bone-cement and bone-wax, respectively. The authors found (1) an increased bone-mineral content and an increased periosteal apposition after filling with bone-wax in comparison with the reamed side and (2) a decreased bone mineral content and a decreased periosteal apposition on the cemented side in comparison with bone-wax. These findings are interpreted as consequential upon repair processes with a high remodeling and mineralization rate after reaming or filling with bone-wax, whereas remodeling in inhibited after filling with acrylic bone-cement. PMID- 8246003 TI - Breakage of a Total Condylar III knee prosthesis. A case report. AB - The authors report the failure of a Total Condylar III (Cintor, Johnson and Johnson, Raynham, MA) knee arthroplasty, implanted in a second revision knee surgery, due to fracture of the tibial spine. The tibial component was in varus position and externally rotated relative to the tibia. This imparted torque and resulted in a spiral fracture of the polyethylene spine. This is the first report of failure of a Total Condylar III prosthesis by mechanical breakage. PMID- 8246004 TI - Femoral head autografts and contralateral total hip arthroplasty. PMID- 8246005 TI - Changes in muscular proteins during simulated microgravity. PMID- 8246006 TI - Behcet's disease and Sjogren syndrome in a patient with HTLV-I-associated myelopathy. PMID- 8246007 TI - A histometrical study on the globus pallidus in Huntington's disease. AB - Pathological change of the globus pallidus (GP) in 6 cases of Huntington's disease (HD) was examined histometrically by comparison with 10 normal control cases. All but 1 case of HD were in late stages of the disease. Total neuronal count, area of GP, and neuronal cell density were measured in 5 selected regions of coronal sections taken along the antero-posterior axis. Contrary to the findings of previous reports, no neuronal depletion was recognized in HD in any region despite marked atrophy of tissue bulk. The atrophy was more severe in the external segment (GPe) than in the internal segment (GPi). Reactive astrocytosis and fibrillary gliosis were observed in the atrophic lesions. These results indicate that atrophy of the GP can be attributed to striato-pallidal fiber loss and not to neuronal depletion even in the late stages. These findings support the hypothesis that loss of striato-GPe fibers plays the most important role in choreic movements in HD. It remains to be determined whether the pallidal neurons are also preserved in the end stage of the disease. PMID- 8246008 TI - A fine structural study of Onuf's nucleus in sporadic amyotrophic lateral sclerosis. AB - Onuf's nucleus of the second sacral spinal cord level was examined in twelve patients with sporadic amyotrophic lateral sclerosis (ALS). Under light microscopic examination, the neurons of Onuf's nucleus were well-preserved in contrast with the severe neuronal depletion of the anterior horn cells. Ultrastructurally, however, substantial pathological changes such as Bunina bodies (BB), randomly arranged filaments associated with fine granules corresponding to Lewy body-like hyaline inclusions (LBHI) and bundles of tubular filaments corresponding to skein-like inclusions (SI) all of which are characteristic features of ALS, were demonstrated ubiquitously in the Onuf's nucleus in eleven of twelve patients. These changes may be of a reactive nature and represent an initial change of neuronal degeneration, since no definite cell destruction was observed in the nucleus. The changes in the Onuf's nucleus correlated with the changes observed in the anterior horns, although to a lesser degree. The more BB, LBHI and SI were observed in the anterior horns of patients with ALS, the more these inclusions were observed in Onuf's nucleus. Thus, we conclude that Onuf's nucleus is also affected by pathological changes in ALS. PMID- 8246009 TI - Intrafamilial variability in dystrophin abundance correlated with difference in the severity of the phenotype. AB - In Duchenne muscular dystrophy, the progression of the disease is always severe and predictable, while in Becker dystrophy there is a wide variability (intra and inter familial) in the severity of the phenotype. We report here a family in which the proband, who is currently 15 years old, is showing a severe DMD progression, while his affected maternal uncle, aged 29, has a more benign course, compatible with BMD. No DNA deletion was detected in both patients. Dystrophin analysis through immunofluorescence and western blotting showed a negative pattern in the youngest patient and a positive one in the oldest. Apparently, this is the first report on intrafamilial variability in dystrophin abundance correlated with a difference in the severity of the phenotype. PMID- 8246010 TI - Reciprocal expression of dystrophin and utrophin in muscles of Duchenne muscular dystrophy patients, female DMD-carriers and control subjects. AB - We examined muscle biopsies from patients with Duchenne muscular dystrophy (DMD: 39 patients) and Becker muscular dystrophy (BMD: 11 patients), female DMD carriers (4 patients), and control subjects (26 persons) for the expression of dystrophin and utrophin. Control subjects showed all fibers to be dystrophin positive, while utrophin staining was negative or weak. On the other hand, muscles from DMD patients showed the inverse staining patterns: dystrophin was negative and utrophin staining strong. Thus, there was a reciprocal pattern of expression between dystrophin and utrophin. This reciprocal relationship was confirmed to some extent at the single-fiber level in female carriers of DMD showing a mosaic immunostaining of dystrophin. We consider that utrophin may have a function similar to that of dystrophin, and compensate to some extent for dystrophin deficiency in DMD. PMID- 8246011 TI - Abnormal localization of laminin subunits in muscular dystrophies. AB - To address potential involvement of muscle basal lamina and membrane cytoskeleton proteins in the etiology of non-dystrophinopathy muscular dystrophies, we examined the immunostaining intensity and distribution of laminin subunits (A, B1, B2 and M), type IV collagen, dystrophin and spectrin in skeletal muscle biopsies from 64 myopathic patients (17 Fukuyama congenital muscular dystrophy: FCMD, 13 congenital muscular dystrophy unrelated to FCMD: other CMD, 16 Duchenne muscular dystrophy: DMD, and 18 other neuromuscular diseases. In FCMD muscle, we found a significant reduction of laminin M (merosin; a striated muscle specific basal lamina-associated protein) with approximately 26% of levels seen in controls by quantitative immunofluorescence. Other CMD and DMD muscles showed less dramatic reductions (78%, 80%, respectively). The localization of laminin M was also abnormal in FCMD muscle. Laminin B1 and B2 showed abnormalities similar to those observed with laminin M, but were less marked. Laminin A was only detected in rare regenerating fibers in control biopsies, whereas it was seen around most muscle fibers in FCMD patients, and in dystrophin deficient muscle fibers from DMD patients and its carrier. Staining intensity of type IV collagen in FCMD muscle was not significantly different from the other diseases. These findings may implicate a primary or central role for the basal lamina in FCMD muscle. PMID- 8246012 TI - Further impairment of muscle phosphate kinetics by lengthening exercise in DMD/BMD carriers. An in vivo 31P-NMR spectroscopy study. AB - We used phosphorus magnetic resonance spectroscopy (31P-MRS) to study the effect of exercise-induced muscle injury in the calf muscle of 7 DMD/BMD carriers and 6 non-carrier females. All subjects performed 50-80 lengthening contractions with the right calf muscles. 48 h after lengthening exercise non-carriers showed increased sensitivity to pressure in their gastrocnemius accompanied by increased T2 relaxation times and by elevated Pi/PCr ratios at rest. DMD/BMD carriers did not show any effect of lengthening exercise on these measurements. In-magnet exercise revealed in all carriers a reduced initial rate of Pi recovery and an increased time to fully recovery the resting value of intracellular pH. Lengthening exercise further decreased the initial rate of Pi recovery. Non carriers did not show any variation attributable to lengthening exercise either during in-magnet work or during recovery from exercise. We found that lengthening exercise contractions causes: (1) less muscle injury in carriers compared to non carriers, (2) even slower rate of Pi recovery, but (3) no effect on Pi recovery in non-carriers. The use of lengthening exercise and measurements of Pi recovery may be a useful method to evaluate the disease process in DMD/BMD. PMID- 8246013 TI - Collagen fibril diameter and its relation to cross-linking of collagen in the skin of patients with amyotrophic lateral sclerosis. AB - The diameter of collagen fibrils and the age-related, stable cross-link, histidinohydroxylysinonorleucine (HHL), of skin tissue (left biceps) were measured and compared in 11 patients with amyotrophic lateral sclerosis (ALS) and 7 controls. In ALS patients, it was found that (1) the diameter of collagen fibrils and the content of HHL were negatively and significantly associated with the duration of illness (r = -0.74, P < 0.01 and r = -0.72, P < 0.02, respectively), and (2) there was an appreciable positive correlation between the diameter of collagen fibrils and the content of HHL (r = 0.73, P < 0.02). These findings were not observed in controls. These suggest there is a close relationship between the decreased amount of HHL and the small diameter of collagen fibrils in ALS. PMID- 8246014 TI - Swelling and death of neuronal cells by lactic acid. AB - Lactacidosis occurring in cerebral ischemia or trauma is a major mechanism of cytotoxic brain edema and brain damage. Respective effects of lactacidosis were currently analyzed in vitro by employment of the murine neuronal cell line, Neuro 2A, in order to obtain a better understanding of specific mechanisms underlying cell swelling and cell death in comparison with glial cells. The cells were suspended in a physiological medium in the presence of lactic acid at increasing concentrations. Levels of acidosis reaching from pH 6.8-5.6 were obtained while other parameters, such as osmolarity and electrolyte concentrations, were maintained in the physiological range. Assessment of cell swelling and cell viability using exclusion of propidium iodide was made by flow cytometry with employment of an advanced Coulter system. Swelling of Neuro-2A cells commenced once the pH in the medium was lowered to 6.8 or below. From this level downward, cell swelling was a function of the severity of acidosis and duration of exposure. For example, lactacidosis of pH 6.8 or 5.6 lasting 90 min led to an increase in cell volume to 109.5% or 159.6% of normal, respectively. Viability of the neuronal cells was 85% under control conditions. It remained in this range down to pH 6.2. At pH 5.6, however, cell viability decreased in a time-dependent fashion. At 90 min, only 48.9% of the neuronal cells were viable at pH 5.6. The swelling response and impairment of viability of the neuronal cells was compared with that of C6 glioma cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246015 TI - Transient ischemic attacks: electrophysiological (conventional and topographic EEG) and radiological (CCT) evaluation. AB - The value of electrophysiological tests: conventional electroencephalography (EEG), topographic EEG analysis as well as computerized tomography (CT) in the diagnosis and evaluation of 25 patients with manifestations of transient ischemic attacks (TIA) in the domain of the carotid system was assessed. Normal CT was the rule in TIA patients except in 8% of the cases, where nonspecific changes of brain atrophy were described. Conventional EEG, topographic EEG and spectral analysis could detect abnormalities in 48%, 80% and 64% of TIA cases respectively. None of the abnormal EEG records could be missed by topographic EEG analysis. 32% of the records were diagnosed as abnormal by topographic EEG, while conventional EEG failed to detect abnormalities. Spectral analysis of the EEG results revealed a significant decrease regarding mean high limit alpha percent power, and a significant increase regarding mean low and high limit theta percent power, as well as a significant increase of the mean high limit of the slow activities (delta + theta)/fast activities (alpha + beta) percent power ratio in the TIA group as compared to the normal control group. PMID- 8246016 TI - Copper-64 metabolism in two patients with non-Wilsonian movement disorders and copper deficiency. AB - Copper-64 studies are presented of 2 patients with non-Wilsonian movement disorder and with abnormal copper handling. Both patients differed from the usual phenotype of non-Wilsonian low copper movement disorder as they had choreiform movement disorders with an onset in the first decade; one patient lacked significant intellectual impairment. Both patients had reduced serum total copper and marginal free copper and caeruloplasmin levels, and both patients were capable of incorporating 64Cu2+ into caeruloplasmin but the second case did so at markedly reduced level. Both showed slightly increased basal and stimulated urinary copper loss compared to normal controls with the rate in patient 1 being capable of leading to copper depletion. Liver copper content was normal in both cases. These 2 patients add to the reports of cases with copper deficiency and movement disorder in whom copper chelation therapy is unlikely to be beneficial. PMID- 8246017 TI - Primary large-cell lymphoma of the mediastinum. PMID- 8246018 TI - Pharmacologic-based dosing of carboplatin: a better method. PMID- 8246019 TI - Thoughts from a journey. PMID- 8246020 TI - Primary mediastinal B-cell lymphoma with sclerosis: an aggressive tumor with distinctive clinical and pathologic features. AB - PURPOSE: To evaluate the clinical features of presentation, the morphologic and immunohistochemical pattern, the modality of spread, and the response to current treatments of patients with primary mediastinal B-cell lymphoma, a recently documented subtype of non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Thirty consecutive patients (14 males, 16 females; median age, 26 years) with primary mediastinal B-cell lymphoma with sclerosis were studied. RESULTS: The clinical aspects were largely homogeneous: 93% presented with chest symptoms of a rapidly enlarging mass of the anterior mediastinum; the tumor was bulky in 73%, and superior vena cava syndrome (SVCS) was present in 57%. Also, patients without SVCS symptoms showed subclinical venacaval compression at computed tomographic (CT) scan, for a total incidence of caval obstruction of 80%. Intrathoracic extension to adjacent organs was seen in 47% of patients. Despite its invasive behavior, only four patients showed extrathoracic spread at diagnosis. In 23 cases, the tumor presented with morphologic features that resembled follicular center-cell lymphomas. In seven, the neoplastic population was composed mainly of centrocyte-like cells with abundant clear cytoplasm not referable to any known B cell lymphoma subtype. All cases showed huge sclerosis. Of 29 patients assessable for response, 16 (55%) achieved a complete response (CR): five of 14 (36%) treated with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), and 11 of 15 (73%) treated with methotrexate plus leucovarin, doxorubicin, cyclophosphamide, vincristine, prednisone, and bleomycin (MACOP-B) or etoposide, doxorubicin, cyclophosphamide, vincristine, prednisone, and bleomycin (VACOP-B) (P = .047). We could identify no clinical, biologic, or histopathologic features significantly correlated with response. After chemotherapy, 14 of 16 remitters received consolidation radiotherapy to the mediastinum. At 3 years, the actuarial survival rate is 38% for all cases and 72% for remitters. None of the 13 patients who did not achieve CR responded to salvage treatments. CONCLUSION: This study shows that primary mediastinal B-cell lymphoma with sclerosis is a distinctive subtype of NHL with unique clinicopathologic aspects and aggressive behavior. Prompt recognition and aggressive treatment may provide long survival in a good proportion of cases. However, a subset of patients is extremely refractory to first- and second-line treatment. Conventional prognostic factors seem inadequate to identify these very-poor-risk cases. PMID- 8246021 TI - Carboplatin pharmacokinetics in children: the development of a pediatric dosing formula. The United Kingdom Children's Cancer Study Group. AB - PURPOSE: The aim of this study was to define the pharmacokinetics of carboplatin in children and use the data to develop a pediatric dose formula. It was anticipated that renal function would be a major determinant of carboplatin disposition and the relationship between carboplatin clearance and glomerular filtration rate (GFR) was examined in detail. PATIENTS AND METHODS: Plasma carboplatin pharmacokinetics were measured as ultrafiltrable platinum in 22 patients (5 to 63 kg) following 200 to 1,000 mg/m2 of carboplatin. GFR was measured by the plasma clearance of chromium 51-edathamil (51Cr-EDTA). RESULTS: Carboplatin pharmacokinetics in children were best described in most patients (16 of 22) by a two-compartment model. The dose-normalized area under the plasma carboplatin concentration versus time curve (AUC) ranged from 3.1 to 9.6 mg/mL.min/400 mg/m2 and there was only a weak linear relationship between carboplatin dose and AUC (R2 = .31). There was a significant relationship between absolute carboplatin and 51Cr-EDTA clearances (R2 = .56), but the relationship was weaker (R2 = .28) when both clearances were normalized for body surface area. Carboplatin plasma clearance was predicted by the equation: clearance = GFR (mL/min) + 0.36 x body weight (BW; kg), and a modified form of the adult carboplatin dose formula is proposed: dose (mg) = target AUC x (GFR [mL/min] + [0.36 x BW(kg)]). Two further equations were developed that use the 51Cr-EDTA half-life (t1/2) to calculate the GFR and these may reduce errors resulting from inaccurate measurement of the volume of distribution for 51Cr-EDTA. In patients treated with single-agent carboplatin or carboplatin plus vincristine, there was a significant sigmoidal relationship between AUC and thrombocytopenia (R2 = .56). CONCLUSION: GFR-based carboplatin dosing in children should be feasible and will be evaluated prospectively. PMID- 8246022 TI - Impact of megatherapy on survival after relapse from stage 4 neuroblastoma in patients over 1 year of age at diagnosis: a report from the European Group for Bone Marrow Transplantation. AB - PURPOSE: Relapse from stage 4 neuroblastoma usually carries a poor prognosis. A retrospective study using the European Bone Marrow Transplant (EBMT) Solid Tumor Registry was undertaken to define the role of megatherapy (MGT) in relapsed patients. PATIENTS AND METHODS: After relapse, 33 boys and 15 girls with previous stage 4 neuroblastoma received intensification by MGT followed by either autologous (n = 42) or allogeneic (n = 6) bone marrow rescue in 11 European institutions. The median age at diagnosis was 47 months (range, 14 to 134) and the median interval from diagnosis to relapse was 16 months (range, 4 to 94). Thirty patients had received only conventional-dose primary treatments (group A), whereas 18 patients had previously received intensification with MGT (group B). The median follow-up time of the total group is 95 months (range, 25 to 185). RESULTS: The actuarial overall survival rate at 2 years after MGT for relapse is 27% for group A and 0% for group B (P = .02). Three adverse, independent prognostic factors were confirmed by multivariate analysis using the Cox proportional hazards regression model: an interval of less than 12 months between diagnosis and relapse (P < .0001), nonresponding or untreated relapse (P = .0002), and previous MGT during primary treatments (P = .055). None of the other variables analyzed, such as sex, age, bone or bone marrow involvement at diagnosis or at relapse, and type of MGT at relapse, influenced outcome in this patient cohort. CONCLUSION: Responding patients who relapse more than 12 months from diagnosis who had not received previous MGT appear to benefit from consolidation MGT. Relapse patients who do not fulfill these criteria gain no advantage from this cost-intensive procedure and should be treated differently. PMID- 8246023 TI - Allogeneic, syngeneic, and autologous marrow transplantation for Hodgkin's disease: the 21-year Seattle experience. AB - PURPOSE: To analyze results of 127 patients undergoing myeloablative therapy followed by marrow transplantation for relapsed or refractory Hodgkin's disease. PATIENTS AND METHODS: Twenty-three patients had primary refractory disease, 34 were in early first relapse or second complete remission (CR), and 70 had refractory first relapse or disease beyond second CR. Preparative regimens included total-body irradiation (TBI) and chemotherapy (n = 61) or chemotherapy only (n = 66). Sixty-eight patients received autologous marrow, six syngeneic marrow, and 53 allogeneic marrow. RESULTS: The 5-year actuarial probabilities of survival, event-free survival (EFS), relapse, and nonrelapse mortality for the entire group were 21%, 18%, 65%, and 49%, respectively. HLA-identical allogeneic marrow recipients had a statistically lower relapse rate compared with recipients of autologous marrow, but survival, EFS, and nonrelapse mortality rates were not significantly different. In the multivariate analysis, higher performance status and absence of bulky disease predicted for improved EFS and lower relapse rates, while fewer prior treatment regimens predicted for improved EFS and lower nonrelapse mortality rates. Additionally, the univariate analysis showed that patients who underwent transplantation with disease refractory to chemotherapy or beyond second CR had a worse outcome compared with those who had less advanced disease. CONCLUSION: Outcome with transplantation for patients with Hodgkin's disease is improved if transplantation is performed early after relapse when disease burden is less, tumor chemosensitivity is greater, and the patient is likely to have a better performance status. The use of HLA-matched sibling marrow results in a lower relapse rate and, thus, for some individuals, may be preferable to the use of autologous marrow. PMID- 8246024 TI - One hundred autotransplants for relapsed or refractory Hodgkin's disease and lymphoma: value of pretransplant disease status for predicting outcome. AB - PURPOSE: One hundred autotransplants for Hodgkin's disease (HD) or non-Hodgkin's lymphoma (NHL) were examined prospectively to identify variables with prognostic significance. PATIENTS AND METHODS: Ninety-six patients with relapsed or refractory HD or NHL underwent 100 autotransplants. Patients received high-dose carmustine (BCNU), etoposide, cytarabine, and cyclophosphamide (BEAC) followed by unpurged autologous stem-cell rescue. RESULTS: The 3-year actuarial event-free survival (EFS) rate for the 47 HD patients is 49%, with a median followup duration of 2 years. For the 53 NHL patients, the 3-year actuarial EFS rate is 40%, with a median follow-up duration of 19 months. By multivariate analysis, minimal disease on admission (all areas < or = 2 cm) is associated with improved EFS (HD, P = .003, NHL, P = .03). The projected EFS rate for HD patients entering with minimal disease is 70% versus 15% for patients with bulky disease (P = .0001). The projected EFS rate for NHL patients with minimal disease is 48% versus 25% for patients with bulky disease (P = .04). Posttransplant involved field radiotherapy, administered to 26 of the last 61 patients, was associated with an improved EFS rate for NHL patients (P = .015). The BEAC regimen was well tolerated by patients who entered the study with minimal disease (mortality rate, < 5%), but caused significant toxicity in patients with bulky disease (mortality rate, 25%). CONCLUSION: Disease burden before autotransplantation is an important predictor of regimen-related toxicity and EFS. Posttransplant involved-field radiotherapy may improve outcomes in select patients with NHL. The BEAC regimen is safe and effective, particularly for patients with minimal disease. PMID- 8246025 TI - Therapy-related acute myeloid leukemia with t(8;21), inv(16), and t(8;16): a report on 25 cases and review of the literature. AB - PURPOSE: To analyze therapy-related acute myeloid leukemias (tAMLs) with t(8;21), inv(16), or t(8;16). PATIENTS AND METHODS: Twenty-five patients with tAML and t(8;21)(q22;q22), inv(16)(p13;q22), or t(8;16)(p11;p13) from seven centers, along with 23 previously published cases, were studied. RESULTS: Twenty-six, 16, and six patients, respectively, had t(8;21), inv(16), and t(8;16). Prior cancer was a solid tumor in 27 cases, and a hematologic malignancy in all other patients. Five patients had received prior radiotherapy (RT) alone, and 43 had received prior chemotherapy with or without RT. Prior chemotherapy included a drug that directly reacts with DNA (alkylating agent or cisplatin) and/or an agent that targets topoisomerase II (ATTop, an anthracycline or derivative or, less often, epipodophyllotoxin) in most patients. The interval between prior tumor and diagnosis of tAML was less than 3 years in most cases, and only seven patients had a preleukemic phase of disease. Morphology was M2 AML for t(8;21), M4eo for inv(16), and M4 or M5 for t(8;16). Sixteen of 21 (76%), 12 of 14 (86%), and zero of four patients with t(8;21), inv(16), and t(8;16), respectively, achieved complete remission (CR) with intensive chemotherapy. The actuarial disease-free survival rate at 24 months was 47% and 54% in patients with t(8;21) and inv(16), respectively. CONCLUSION: Like other tAMLs with a karyotype specific of de novo AML [balanced 11q23 rearrangement or t(15;17)], tAMLs with t(8;21), inv(16), or t(8;16) are usually characterized by a short latent period, previous treatment often combining a drug that directly reacts with DNA and an ATTop, and absence of preleukemic phase. Hematologic characteristics and response to treatment are also identical to those of de novo AML with the same karyotypes. PMID- 8246026 TI - Preoperative endoscopic ultrasound can predict the risk of recurrence after operation for gastric carcinoma. AB - PURPOSE: Endoscopic ultrasonography (EUS) has been shown to determine accurately the depth of invasion of the stomach wall in gastric carcinoma. We undertook this study to determine if T (tumor) stage as determined by EUS correlated with recurrence after resection and could be used to identify patients preoperatively at high risk for recurrence. MATERIALS AND METHODS: We reviewed the surgical pathology and obtained follow-up data from the first 50 patients who underwent preoperative EUS for staging of gastric carcinoma. Rotating sector-scan ultrasound endoscopes were used with switchable frequencies of 7.5 MHz and 12 MHz. RESULTS: Of 50 patients, 43 underwent resection with curative intent and were available for follow-up. The concordance of EUS T stage with pathologic T stage in these patients was 86%. At a median follow-up duration of 25 months, only two of 13 patients with preoperative EUS stage T1 or T2 disease were found to have recurrence, while 23 of 30 patients with EUS stage T3 or T4 disease had recurrence (P = .0002) and 22 died. CONCLUSION: We conclude that patients with a preoperative EUS stage T1 or T2 are at low risk for postoperative recurrence, while patients with EUS stage T3 or T4 are at high risk for early postoperative recurrence. The latter patients are reasonable candidates for controlled trials of alternative preoperative management programs, such as chemotherapy, in an effort to improve their poor prognosis. PMID- 8246027 TI - Hepatic toxicity associated with fluorouracil plus levamisole adjuvant therapy. AB - PURPOSE: To determine the frequency and nature of hepatic toxicity associated with fluorouracil (5-FU) plus levamisole adjuvant therapy. PATIENTS AND METHODS: All patients had resection of stage II or stage III colon cancer and were randomized to receive observation only, levamisole alone, or 5-FU plus levamisole. Serial liver function studies were documented in 1,025 patients who did not develop recurrence during the year of therapy. RESULTS: One hundred forty nine (39.6%) of 376 patients treated with 5-FU plus levamisole showed laboratory abnormalities consistent with hepatic toxicity, compared with 16.3% of 251 patients treated with levamisole alone and 16.1% of 398 untreated controls. Most common was elevation of alkaline phosphatase, frequently accompanied by elevations of transaminase or serum bilirubin. Characteristically, these changes were mild, not associated with symptoms, and resolved when therapy was stopped. In some instances, they were associated with elevated carcinoembryonic antigen (CEA) tests or with fatty liver seen on computed tomographic (CT) scan or liver biopsy. CONCLUSION: Mild and reversible hepatotoxicity is a common consequence of 5-FU plus levamisole adjuvant therapy, but this is only rarely symptomatic. However, the oncologist should be alert to this phenomenon, since the associated laboratory and imaging findings may simulate those associated with hepatic metastasis. PMID- 8246028 TI - Ondansetron versus ondansetron, dexamethasone, and chlorpromazine in the prevention of nausea and vomiting associated with multiple-day cisplatin chemotherapy. AB - PURPOSE: This study is designed to evaluate the effectiveness of ondansetron alone (OND) or in combination with 2 days of dexamethasone and 5 days of chlorpromazine (ODC) in the prevention of emetic episodes in patients receiving multiple-day cisplatin. PATIENTS AND METHODS: Forty-four patients receiving 20 or 40 mg/m2 of cisplatin daily for 4 to 5 days plus etoposide (VP-16) alone or in combination with bleomycin or ifosfamide were randomized to receive three doses of OND (0.15 mg/kg 30 minutes before and 4 and 8 hours after cisplatin) versus the identical OND regimen plus dexamethasone 8 mg before cisplatin and 4 mg 4 and 8 hours later on days 1 and 2, plus chlorpromazine 50 mg every 4 hours for four doses per day. Patients were chemotherapy-naive, had a Karnofsky performance status > or = 60, and were not receiving nonstudy antiemetics. RESULTS: Nineteen of 22 patients (86%) on ODC had fewer than three emetic episodes throughout the study period, compared with 10 of 22 (46%) on OND (P = .009), and 55% of patients on ODC had no emetic episodes, compared with 32% on OND (P = .22). The ODC arm had fewer treatment failures (5%) than the OND arm (32%). The mean nausea ratings per visual analog scale were 15.0 for OND and 5.5 for ODC (P = .046). Headache was less frequent with ODC versus OND (14% and 41%, respectively, P = .09). CONCLUSION: ODC was superior to OND with respect to therapeutic efficacy and decreased headaches. Both OND and ODC were more effective on days 1 and 2, rather than days 4 and 5, suggesting tachyphylaxis, anticipatory nausea, or delayed nausea from the first few days of cisplatin combination chemotherapy. PMID- 8246029 TI - Difference in persistence of efficacy of two antiemetic regimens on acute emesis during cisplatin chemotherapy. The Italian Group for Antiemetic Research. AB - PURPOSE AND METHODS: We conducted a prospective, double-blind, multicenter study of 287 cancer patients treated for the first time with high-dose cisplatin chemotherapy who were randomly assigned to receive three consecutive cycles of the same antiemetic treatment consisting of ondansetron plus dexamethasone, or metoclopramide plus dexamethasone and diphenhydramine. RESULTS: Patients who received the ondansetron combination achieved significantly greater complete protection from vomiting, but not from nausea, in all three cycles of chemotherapy than did patients treated with metoclopramide (78.7% v 59.6%, P < .002 during the first cycle; 73.4% v 51.0%, P < .002 during the second cycle; 73.7% v 47.5%, P < .001 during the third cycle). The ability of ondansetron treatment to protect patients from vomiting during the first cycle did not change in subsequent cycles, but decreased significantly as far as complete protection from nausea and from both nausea and vomiting are concerned. With the metoclopramide combination, a significantly greater reduction of complete protection from vomiting, nausea, and both nausea and vomiting was detected. Protection obtained in previous cycles of chemotherapy was the most important prognostic factor. Adverse events were significantly less frequent with ondansetron treatment during the three cycles of chemotherapy and no cumulative toxic effects were found with either treatment. CONCLUSION: Ondansetron plus dexamethasone was significantly more efficacious and better tolerated than metoclopramide plus dexamethasone and diphenhydramine during three cycles of chemotherapy and, in contrast to the metoclopramide regimen, the efficacy of ondansetron plus dexamethasone, at least for vomiting, is maintained in subsequent cycles. PMID- 8246030 TI - Phase I trial of recombinant human granulocyte-macrophage colony-stimulating factor derived from yeast in patients with breast cancer receiving cyclophosphamide, doxorubicin, and fluorouracil. AB - PURPOSE: To establish the optimum biologic dose and maximal-tolerated dose (MTD) of once-daily, subcutaneous recombinant human granulocyte-macrophage colony stimulating factor derived from yeast (RhuGM-CSF) in patients with breast cancer. PATIENTS AND METHODS: Seventeen patients with either newly diagnosed breast cancer with more than four involved axillary nodes (five patients) or metastatic breast cancer (12 patients) were treated with cyclophosphamide 1 g/m2, doxorubicin 50 mg/m2, and fluorouracil 500 mg/m2 (CAF) intravenously (IV) once every 3 weeks. RhuGM-CSF was administered subcutaneously once daily for 14 days after the second and third CAF cycles, at one of three dose levels. RESULTS: The 125-micrograms/m2/d RhuGM-CSF dose level shortened the duration of neutropenia in only one of three patients. The 250-micrograms/m2/d level was effective in shortening the duration of the neutropenic nadir (< .5 x 10(9)/dL) by 2 or more days in five of six patients. The 500-micrograms/m2/d level caused severe toxicity (chest pain, two patients; deep vein thrombosis, one patient) in three of eight patients. CONCLUSION: RhuGM-CSF administered once daily at the 250 micrograms/m2/d level is well tolerated and effective in shortening the duration of the neutrophil nadir by 2 or more days after CAF therapy. PMID- 8246031 TI - Phase I/pharmacokinetic study of high-dose progesterone and doxorubicin. AB - PURPOSE: We developed a new formulation of progesterone that permits administration of up to 10 g of progesterone as a continuous intravenous infusion over 24 hours and conducted a phase I clinical trial to determine whether progesterone could modulate the in vivo cytotoxicity of the P-glycoprotein substrate doxorubicin. PATIENTS AND METHODS: Thirty-four patients with advanced malignancies were treated with increasing doses of progesterone and a fixed dose of 60 mg/m2 of doxorubicin given as an intravenous bolus 2 hours after starting a 24-hour intravenous infusion of progesterone. RESULTS: Progesterone enhanced doxorubicin-induced myelotoxicity in a dose-dependent fashion without altering the pharmacokinetics of doxorubicin. The steady-state plasma concentration of progesterone at a dose level of 4 g was 4.1 +/- 0.9 mumol/L, which was higher than the minimal concentration required to reverse multidrug resistance (MDR) in vitro. CONCLUSION: Progesterone enhanced the hematologic toxicity of doxorubicin without altering its pharmacokinetics, suggesting that progesterone could modulate P-glycoprotein at the level of pluripotent hematopoietic stem cells. Adequate tissue concentrations of progesterone could be achieved in vivo to modulate doxorubicin toxicity in the bone marrow and thus potentially in tumor tissue as well. Selectivity may potentially be gained by using hematopoietic growth factors to offset the enhanced hematologic toxicity of doxorubicin while leaving the enhancement of toxicity to tumor cells unchanged. PMID- 8246032 TI - Leukemoid reaction, bone marrow invasion, fever of unknown origin, and metastatic pattern in the natural history of advanced undifferentiated carcinoma of nasopharyngeal type: a review of 255 consecutive cases. AB - PURPOSE: This study is an analysis of frequency and relationship regarding two undifferentiated carcinoma of nasopharyngeal type (UCNT)-associated paraneoplastic syndromes (PNS): leukemoid reaction (LR) and fever of unknown origin (FUO) with bone marrow invasion (BMI) and metastatic pattern. PATIENTS AND METHODS: A consecutive UCNT patient cohort (N = 255) with locally advanced (n = 142) or metastatic (n = 113) disease receiving chemotherapy alone or in combination with radiotherapy was studied. All patients had a complete baseline work-up that included bone marrow biopsy. RESULTS: UCNT has distinctive features among head and neck squamous cell cancers (HNSCC). These include early subclinical dissemination, with 70% of metastases appearing within 18 months of first symptoms. Metastases are common in bone (65% v 25% in HNSCC), liver (29% v 23%), and lung (18% v 84%), and BMI is observed in 25% of UCNT patients with metastases. Metastases likelihood is related to lymph node involvement, with 64% of patients with metastases having N3 disease. Involved lymph nodes in contrasted CT scans revealed hypodensity in 26% of UCNT patients versus 79% in patients with other HNSCC. Hypercalcemia was observed in one case. LR was identified in 41 patients (16%); in 26 of the 41 patients (64%) it was observed concomitant with N3 and/or metastatic disease. FUO was found in 23 patients (9%) and was associated in four instances with BMI and in 17 with LR (in four instances with both). Brain metastases or meningeal carcinomatosis were not observed despite the high rate of skull base compromise. Paraneoplastic signs were observed in 47 of 255 cases (18.5%) and were more frequent in patients with metastases. However, PNS were observed in 15 patients with negative metastases work-up. CONCLUSION: The PNS described could help in the diagnosis and follow-up of UCNT patients because they may be the first manifestation of the disease or may reappear with relapse. BMI is a frequent finding in patients with metastases and is unrelated to PNS. PMID- 8246033 TI - Low-dose gallium nitrate for prevention of osteolysis in myeloma: results of a pilot randomized study. AB - PURPOSE: Since osteolysis is a major cause of morbidity in myeloma, we conducted a pilot study to evaluate whether the addition of gallium nitrate to standard antimyeloma treatment could preserve or increase bone mass in patients with osteolytic disease. METHODS: Patients stabilized on cytotoxic therapy were randomized to treatment with gallium nitrate for 6 months, or to observation only for the first 6 months followed by gallium nitrate treatment during the subsequent 6 months. Gallium nitrate was administered in monthly cycles by daily subcutaneous injections (30 mg/m2/d) for 2 weeks, followed by 2 weeks with no therapy, supplemented by an intravenous infusion (100 mg/m2/d) for 5 days every other month. RESULTS: Paired 6-month comparisons were available for seven observation periods and 13 gallium nitrate treatment periods. Total-body calcium assessed by delayed-gamma neutron activation (DGNA) decreased in four of seven patients during observation, but increased in nine of 13 patients during gallium nitrate treatment; the mean difference in total-body calcium (TBCa) between the two groups at 6 months was 3%. Median regional bone density assessed by dual photon absorptiometry (DPA) declined by 1.4% in patients under observation (range, +6.7% to -18.3%), but was unchanged during gallium nitrate treatment (median change, 0%; range, -10.5% to +14.4%). The group mean vertebral fracture index assessed by lateral spine x-rays decreased by 27% during observation compared with 2% during gallium nitrate treatment. Mean body height decreased by 0.57 inches in the observation group and .06 inches in the gallium nitrate group. Patient self-assessment of bone pain showed that seven of 12 gallium nitrate treated patients rated themselves as experiencing major reductions in bone pain, compared with zero of seven patients who were observed. One episode of hypercalcemia occurred in a patient under observation. CONCLUSION: Adjuvant treatment with low-dose gallium nitrate attenuates the rate of bone loss in myeloma and may be useful for ameliorating the consequences of skeletal morbidity in patients with cancer-related osteolysis. PMID- 8246034 TI - Inadequacy of ultrasonography for monitoring response to treatment of liver metastases. AB - PURPOSE: We prospectively evaluated the clinical efficacy of ultrasonography (US) in monitoring the effect of medical treatment in patients with liver metastases, by comparing serial US assessment with serial magnetic resonance imaging (MRI) evaluation and clinical outcome in a group of 41 patients with solid tumors. PATIENTS AND METHODS: Both examinations were performed in patients with metastatic liver disease at the start of a new treatment modality and monthly thereafter for 3 months; close monitoring was prolonged beyond the third month in cases in which there was disagreement between the two techniques and the clinical course was not conclusive. RESULTS: Planned follow-up was completed in 37 cases. There was limited concordance between the two examinations: in 21 cases only (56.8%), US and MRI gave concordant information on the evolution of hepatic metastases; in eight cases, both agreed on progression of disease (PD), in 11 cases on stable disease (SD), and in one case each on partial response (PR) and complete response (CR). In the remaining 16 cases (43.2%), there was disagreement between the two examinations. On the basis of subsequent clinical course, this discrepancy was shown to be due to US inadequacy in 13 cases and to MRI inadequacy in one case; in two cases, the clinical course was not conclusive. The most striking limits of US appeared to be twofold: (1) a progressive appearance, with chemotherapy, of a diffusely inhomogeneous structure of the liver, resulting in obscuration of focal lesions (and a subsequent judgement of CR) in cases in which lesions were, on the contrary, detected at MRI and usually confirmed by subsequent clinical course; and (2) false US-determined PD in cases in which lesions proven at baseline MRI were noted at US only after one to two courses of therapy. CONCLUSION: We conclude that US, which is known to be inaccurate for screening of liver metastases, is unreliable for the follow-up of metastatic liver disease; despite its wide availability, low cost, and noninvasiveness, critical therapeutic decisions should not be made based on the outcome of this test. PMID- 8246035 TI - Informed consent in compliance studies. PMID- 8246036 TI - Heparinized saline to flush Groshong catheters. PMID- 8246037 TI - Dental composites: a review. AB - Dental composite resins are widely used in dental practice and are continually being developed in order to obtain better products. To gain full benefit from these materials, it is important for the clinician to understand their properties. The following is a review of the more common characteristics of composites in current use. PMID- 8246038 TI - Histiocytosis X: evidence for a genetic etiology. AB - Histiocytosis X is a rare disorder with no particular predilection for race, age or sex. Since its discovery by Hand in 1893, the etiology has remained unknown, although viruses, bacteria and genetic factors have been implicated. Familial occurrence of this disease is very rare, and only a handful of such cases have been reported. The present study adds further evidence to support the influence of genetic factors in the etiology of histiocytosis X. PMID- 8246039 TI - Quantitation of circulating immune complexes in patients with chronic periapical lesions. AB - Quantitation of circulating immune complexes (CIC) was done in 45 patients with chronic periapical lesions. The levels were compared with those of age-matched healthy individuals. Both patients with chronic periapical granuloma and periapical cysts showed significantly higher levels of CIC than the controls. This observation indicates that the continuous presence of root canal antigens may cause elevated levels of circulating immune complexes. The possibility of chronic periapical lesions acting as foci of infection is discussed, and the importance of early treatment of these conditions is emphasized. PMID- 8246040 TI - Property variations of stone casts following high-temperature sterilization. AB - The effects of high-temperature sterilization methods (boiling, dry-heating, autoclaving and chemiclaving) on the properties of four types of stone cast were determined. SEM observations were made of the cast surface before and after sterilization. High-temperature sterilization affected the properties of the stone casts. Boiling, dry-heating and autoclaving were considered to be unacceptable methods for sterilizing stone casts before making prosthetic restorations. PMID- 8246041 TI - Preprosthetic surgery for severely atrophic maxilla with poor palatal vault form; report of two cases treated by hard palate compression through palatal vault osteotomy. AB - Two cases of severely atrophic maxilla with poor palatal vault form treated by Kitayama's modified palatal vault osteotomy technique are described. The advantages of this osteotomy technique are outlined in comparison with other surgical methods. PMID- 8246042 TI - Study on high-strength plastic teeth. Tooth discoloration. AB - Conventional plastic teeth (CV teeth) are inferior to porcelain teeth in maintaining an adequate esthetic appearance with wear and discoloration, and thus have a shorter period of durability. Recently, high-strength plastic teeth (HS teeth) have been developed and applied to overcome the wear problems of CV teeth. Since HS teeth made of hard resin are still susceptible to staining with pigments, it has been observed that the esthetics of removable partial dentures made from such plastic teeth are gradually impaired in many patients. To investigate the susceptibility of HS teeth to pigments, we conducted an in vitro study by immersing three types of artificial teeth in three coloring liquids. It was found that the HS teeth tended to be less susceptible to the test pigments than the CV teeth to various degrees. In contrast, they showed markedly stronger susceptibility to the pigments than porcelain teeth. For all three artificial tooth types, daily tooth cleaning with an ultrasonic vibrator had a tendency to reduce the coloration in comparison with their counterparts without ultrasonic cleaning. PMID- 8246043 TI - Expression of ras-P21 and ras gene alteration in pleomorphic adenomas. AB - The expression of ras-P21 protein and gene alteration of the K-ras gene were examined in 12 cases of pleomorphic adenoma of the salivary gland. The tumor cells in all 12 cases were strongly stained for ras-P21 protein by the indirect immunoperoxidase method, suggesting the enhanced expression of ras-P21 protein in pleomorphic adenomas. Analysis of the PCR products for the K-ras gene by temperature-gradient gel electrophoresis (TGGE) revealed that gene alterations such as point mutations occurred in 4 out of 12 pleomorphic adenoma tissues examined. PMID- 8246044 TI - Swinglock removable partial denture and biting forces in partially edentulous patients. Comparison of individual biting forces with and without swinglock removable partial denture. AB - Several studies have shown that swinglock removable partial dentures are effective for stability of the abutment teeth and the surrounding tissues including alveolar bone. This efficacy has been thought to be due to the dynamic structure of the swinglock attachment. Individual biting force was measured to determine the physiological efficacy of teeth splints by use of the attachment. The results showed that individual biting forces with the swinglock removable partial denture were 10-25% higher than those without the denture. Furthermore, statistical analysis showed that the individual biting forces with the swinglock removable partial denture were significantly higher. The swinglock attachment can stabilize partially edentulous dentition by splinting all residual teeth. In addition, when physical forces are applied to the abutment teeth and the artificial teeth, this attachment can deliver and distribute the stress to the other abutment teeth and alveolar mucosa. The present findings suggest that the swinglock attachment augments the ability to withstand physical forces such as those occurring during biting and mastication. PMID- 8246045 TI - Regional odontodysplasia. AB - An unusual case of regional odontodysplasia in an 8-year-old boy is presented. He was referred to our clinic because of failure of tooth eruption on the left side of the maxilla and frequent fever attacks. Treatment consisted of extraction of the affected teeth, and removal of inflamed tissue. PMID- 8246046 TI - The yin and yang of neurological surgery. The 1993 AANS presidential address. AB - The President of the American Association of Neurological Surgeons (AANS) discusses the present socioeconomic milieu, which has created anxiety in the neurosurgical community. The underlying factors are technological advances, hospital-physician relationships, medical liability, quality assurance reviews, and physician reimbursement. It is proposed that neurosurgeons be proactive in the development of health-care reform. PMID- 8246047 TI - Cerebral oxygen metabolism during hypothermic circulatory arrest in humans. AB - Profound hypothermia with circulatory arrest is an important surgical adjuvant that allows protected cessation of cerebral blood flow for a brief period. In seven patients undergoing this procedure, continuous spectroscopic measurement of cerebral hemoglobin oxygen saturation was performed. Circulatory arrest at 18 degrees C was associated with a significant progressive desaturation (p < 0.01) of residual cerebral hemoglobin. Arrest time varied based on operative complexity (range 10 to 65 minutes), and a negative linear correlation between arrest time (y) and oxygen saturation (x) was noted (y = -0.87 x + 64). Five patients whose saturation remained above 35% had no neurological injury attributable to hypoxia. One patient (Hunt and Hess Grade 0) whose saturation fell below 35% had evidence of a global hypoxic injury at postmortem examination. Spectroscopically measured cerebral hemoglobin saturation (cerebral oximetry) may be used to monitor metabolic activity during circulatory arrest. Although the clinical utility of such monitoring cannot be established at this time, the potential may exist to prolong the safe duration of induced circulatory arrest for cerebral protection. PMID- 8246048 TI - Release of beta-endorphin and methionine-enkephalin into cerebrospinal fluid during deep brain stimulation for chronic pain. Effects of stimulation locus and site of sampling. AB - The authors systematically studied the release of the endogenous opioid peptides beta-endorphin and methionine (met)-enkephalin into the cerebrospinal fluid (CSF) during deep brain stimulation in patients suffering from otherwise intractable chronic pain. Nine patients were included in the study; six had stimulation electrodes placed in both the periventricular gray matter (PVG) and the thalamic nucleus ventralis posterolateralis (VLP) and three in the PVG only. Immunoreactivity of beta-endorphin and met-enkephalin (beta-EPir and MEir, respectively) was measured by radioimmunoassays in ventricular and lumbar CSF samples obtained before, during, and after stimulation. Prestimulation concentrations of beta-EPir and MEir were lower in ventricular than in lumbar CSF (6.6 +/- 0.5 vs. 13.7 +/- 1.0 pmol/liter, p = 0.0001, for beta-EPir; 33.6 +/- 5.1 vs. 48.3 +/- 3.2 pmol/liter, p < 0.05, for MEir). Ventricular CSF concentrations of both beta-EPir and MEir increased significantly during PVG stimulation, whereas VPL stimulation was without effect. No changes were seen in lumbar CSF levels of the peptides during stimulation in either site. A significant inverse relationship was found between the "during:before stimulation" ratios of visual analog scale ratings and beta-EPir levels during PVG stimulation. The beta-EPir and MEir concentration during:before stimulation ratios were positively correlated, whereas no correlation was present in prestimulation samples from ventricular or lumbar CSF. High-performance liquid chromatography of ventricular CSF pools obtained during PVG stimulation revealed that major portions of beta EPir and MEir eluted as synthetic beta-endorphin and met-enkephalin, respectively, thus documenting the release of beta-endorphin and met-enkephalin into ventricular CSF during PVG stimulation. The finding of a direct relationship between beta-EPir release and pain alleviation may suggest a role for beta endorphin in the analgesic mechanism of PVG stimulation. PMID- 8246049 TI - Role of angiography following aneurysm surgery. AB - The postoperative angiograms in 66 patients who underwent craniotomy for clipping of 78 cerebral aneurysms were reviewed. Indications for urgent postoperative angiography included neurological deficit or repeat subarachnoid hemorrhage. Routine postoperative angiograms were carried out in the remaining patients. Postoperative angiograms were reviewed to determine the incidence of unexpected findings such as unclipped aneurysms, residual aneurysms, and unforeseen major vessel occlusions. Logistic regression analysis was used to test if the following were factors that predicted an unexpected finding on postoperative angiography: aneurysm site or size; the intraoperative impression that residual aneurysm was left or a major vessel was occluded; intraoperative aneurysm rupture; opening or needle aspiration of the aneurysm after clipping; or development of a new neurological deficit after surgery. Kappa values were calculated to assess the agreement between some of these clinical factors and unexpected angiographic findings. Unexpected residual aneurysms were seen in three (4%) of the 78 occlusions. In addition, three aneurysms were completely unclipped (4%); these three patients were returned to the operating room and had their aneurysms successfully obliterated. There were nine unexpected major vessel occlusions (12%); six of these resulted in disabling stroke and two patients died. Of six major arteries considered to be occluded intraoperatively and shown to be occluded by postoperative angiography, two were associated with cerebral infarction. Logistic regression analysis showed that a new postoperative neurological deficit predicted an unforeseen vessel occlusion on postoperative angiography. Factors could not be identified that predicted unexpected residual aneurysm or unclipped aneurysm. The inability to predict accurately the presence of residual or unclipped aneurysm suggests that all patients should undergo postoperative angiography. Since a new postoperative neurological deficit is one factor predicting unexpected arterial occlusion, intraoperative angiography may be necessary to help reduce the incidence of stroke after aneurysm surgery. With study of more patients or of factors not examined in this series, it may be possible to select cases more accurately for intraoperative or postoperative angiography. PMID- 8246050 TI - Spinal cord injury without radiographic abnormality and Chiari malformation. AB - Spinal cord injury without radiographic abnormality and asymptomatic Chiari I malformation have an unusual coincidence. A young boy who had recently fallen from his porch was transferred to the neurosurgery service with a high cervical central cord syndrome. Careful study demonstrated no radiographic abnormality and, although the patient was previously quite well, magnetic resonance imaging revealed Chiari I malformation. Although expectedly uncommon, reports of three other similar cases support a less than independent relationship between these two processes. All four children, each aged 2 years, were premorbidly asymptomatic and were playing when they fell from low elevations; two were on a couch. All were evaluated by primary authorities 12 to 48 hours before definitive admission, and all had normal plain film examinations. Three of the four children suffered injuries in flexion, the fourth in extension. Three realized a 5-minute to 3-hour delay before the onset of symptoms, and three suffered gradual progression of deficit. Magnetic resonance imaging was the most commonly applied and productive diagnostic medium, demonstrating cerebellar ectopia in three of three cases. Two children were surgically treated, and all achieved at least a functional outcome. Similarities among these cases support a common mechanism of injury, and indicate careful counseling in children with asymptomatic Chiari I malformation and consideration of operative decompression in those children with progressive neurological injury and deficit. PMID- 8246051 TI - Nondiagnostic CT-guided stereotactic biopsies in a series of 407 cases: influence of CT morphology and operator experience. AB - Nondiagnostic biopsies were analyzed in a consecutive series of 407 patients undergoing computerized tomography (CT)-guided stereotactic biopsies. These were categorized as either negative biopsies, when normal tissue or nonspecific pathology was found, or inconclusive, when a definitive diagnosis could not be made although representative tissue was obtained. Nineteen biopsies (4.7%) were negative and 10 (2.4%) were inconclusive, giving an overall nondiagnostic biopsy rate of 7.1% (29 of the 407 cases). Suspected neoplastic masses (390 cases) were classified on the basis of their CT morphology into four groups: Group 1 included purely hypodense nonenhancing masses; Group 2 included isodense nonenhancing masses; Group 3 included ring-enhancing masses; and Group 4 included mixed density enhancing masses. Although a higher proportion of hypodense nonenhancing masses (six of 56, or 10.7%) yielded a negative result, there was no statistically significant difference in the negative biopsy rates for the different CT categories (p = 0.06). The negative biopsy rates for the 6 years of the study, 1987 to 1992 (1987 being an incomplete year) were as follows: 13.3%, 6%, 3.2%, 3%, 5.8%, and 2.7%. There was no significant decrease in the negative biopsy rate as experience with this procedure increased (p = 0.20). A total of eight surgeons independently performed the biopsies. There was no significant difference (p = 0.24) in the negative biopsy rate of the surgeon with the most experience (124 biopsies, 2.4% negative biopsy rate) compared with that of the seven other surgeons combined (283 biopsies; 5.7% negative biopsy rate). These findings suggest that the yield in a stereotactic biopsy is independent of the CT appearance of the mass. Adherence to certain basic principles in patient and target selection will ensure a reasonable percentage of positive yield with stereotactic biopsy procedures even if the surgeon is relatively inexperienced. There does not appear to be a learning curve in the performance of CT-guided stereotactic biopsies. The management of patients with nondiagnostic biopsies is discussed. PMID- 8246052 TI - Surgical management of brain-stem tumors in children: results and statistical analysis of 75 cases. AB - A study was made of 75 children treated between 1970 and 1990, with partial, subtotal, or total removal of three intrinsic and 72 exophytic or surface brain stem tumors. In all cases, the goal of surgery was to remove as much tumor as possible. Extent of removal was defined according to data obtained from postoperative computerized tomography or magnetic resonance imaging, and was considered partial when only a small amount of tumor was removed, subtotal when a few cubic millimeters of tumor was left, and total when no residual tumor was seen on postoperative radiological investigations. An ultrasonic aspirator was used for the 43 most recent operations. Among tumor removals without the aspirator, 24 (75%) were partial, eight (25%) subtotal, and none total; with the use of the aspirator, the number of partial removals decreased to 44.5% while that of subtotal and total removals increased to 32% and 23.5%, respectively. There were 69 gliomas (92%) and 47 benign tumors (62.6%). Forty-nine patients were irradiated postoperatively, and 14 of the 23 patients whose benign tumors were removed totally or subtotally did not undergo irradiation. This study showed that: 1) the overall prognosis of patients with malignant tumors was poor and was not improved by surgery; 2) the survival rate of those with benign tumors was significantly (p < 0.01) lower after partial removal than after total or subtotal removal (52% and 94%, respectively, at 5 years); 3) comparison of means and proportions (Student's and chi-squared tests) between benign and malignant tumors showed a significant difference relating to patient age (p < 0.03), peritumoral hypodensity (p < 0.001), and preoperative duration of symptoms (p < 0.001); 4) stepwise logistic regression analysis confirmed that two of these three variables were related to malignancy: namely, patient age at surgery (p < 0.03) and presence of peritumoral hypodensity (p < 0.001); and 5) routine postoperative irradiation was contraindicated after total or subtotal removal of benign tumors. PMID- 8246053 TI - Correlation of glucose consumption and tumor cell density in astrocytomas. A stereotactic PET study. AB - To determine histological correlates of the variability of glucose consumption in astrocytomas, the authors performed positron emission tomography (PET) with 18F-2 fluoro-2-deoxy-D-glucose (FDG) and matched the PET scans three-dimensionally with computerized tomography scans obtained in a stereotactic frame before biopsy. Ten patients with astrocytomas of World Health Organization Grade 2 or 3 were studied; patients with glioblastomas, oligodendrogliomas, or oligoastrocytomas were excluded from the study to avoid any confounding effects of different cell types and necroses. In samples of pure tumor, glucose consumption correlated significantly with cell density, but not with nuclear polymorphism. It is concluded that tumor cell density is a major determinant of glucose consumption in astrocytomas. The use of PET with FDG may help to locate the highest cell density and thus improve the diagnostic yield of stereotactic biopsy. PMID- 8246054 TI - Central nervous system gangliogliomas. Part 1: Pathology. AB - Histopathological features that suggest the diagnosis of ganglioglioma require, in most cases, confirmation by special stains to distinguish these tumors from other gliomas. For this purpose, immunostaining for synaptophysin, which has previously been shown to selectively label the cell surface of neoplastic ganglion cells, was used to retrospectively examine glioma tumor specimens. Sixty three cases of ganglioglioma were identified. The files of the Division of Neuropathology of New York University Medical Center contained 45 tumors that had been diagnosed as ganglioglioma, of which 42 were verified by synaptophysin; three cases were reclassified, two as astrocytomas and one as a gangliocytic paraganglioma. Thus, a tumor identified as ganglioglioma based on other criteria was likely to be a ganglioglioma. The other 21 cases of gangliogliomas were originally diagnosed as astrocytoma or mixed glioma, but were shown by synaptophysin staining to be gangliogliomas. In some cases the ultimate diagnosis was obtained after radical surgery provided relatively abundant amounts of tissue, thereby limiting sampling errors, in contrast to the biopsies from which the original diagnoses were made. Histopathological review of these cases demonstrated that four features represent important clues to the correct diagnosis: 1) clusters of large cells potentially representing neurons (without such cells the tumor cannot be classified as a ganglioglioma); 2) no perineuronal clustering of the glial cells around the alleged neoplastic neurons; 3) fibrosis (desmoplasia); and 4) calcification. Binucleate neurons, previously suggested to be common in gangliogliomas, were not frequently found in this series, and lymphocytic infiltrates, while common, are so often found in other tumors that they gave no specific hint that any single neoplasm was a ganglioglioma. The glial elements were astrocytic in all cases, except that one tumor also had oligodendroglial and ependymal patterns. Four tumors also had small mature neurons, as seen in neurocytomas. Cells from one tumor were successfully grown in short-term tissue culture; the culture contained large dividing neurons with synaptophysin immunoreactivity as well as smaller dividing cells, demonstrating that the neuronal cells are a proliferating element in gangliogliomas. PMID- 8246055 TI - Central nervous system gangliogliomas. Part 2: Clinical outcome. AB - The records of 58 patients with gangliogliomas surgically treated between January 1, 1980, and June 30, 1990, were retrospectively reviewed in order to determine long-term survival, event-free survival, and functional outcome resulting after radical resection and to assess the impact of histological grading on outcome. Tumors were located in the cerebral hemisphere in 19 cases, the spinal cord in 30, and the brain stem in nine. Forty-four patients had gross total resection and 14 had radical subtotal resection. Only six patients underwent postoperative irradiation or chemotherapy and, therefore, the outcome was generally related to surgery alone. Of the 58 gangliogliomas, 40 were classified as histological grade I, 16 were grade II, and two were grade III. The median follow-up period was 56 months. There were no operative deaths, and the operative morbidity rate was 5%, 37%, and 33% for cerebral hemisphere, spinal cord, and brain-stem gangliogliomas, respectively. The 5-year actuarial survival rates for cerebral hemisphere, spinal cord, and brain-stem gangliogliomas were 93%, 84%, and 73%, respectively (p = 0.7). The event-free survival rate at 5 years was 95% for cerebral hemisphere gangliogliomas and 36% for spinal cord gangliogliomas (p < 0.05); for brain-stem gangliogliomas the event-free survival rate at 3 years was 53% (p < 0.05). Neurological function at recent follow-up evaluation was stable or improved in 81% of patients. Multivariate analysis (Cox linear regression) revealed tumor location to be the only variable predictive of outcome, with spinal cord and brain-stem gangliogliomas having a 3.5- and 5-fold increased relative risk of recurrence, respectively, compared to cerebral hemisphere gangliogliomas. Histological grade was not predictive of outcome, although in each location there was a trend for higher-grade tumors to have a shorter time to recurrence. It is concluded that radical surgery leads to long-term survival of patients with gangliogliomas, regardless of location, and adjuvant therapy can probably be reserved for special cases. PMID- 8246056 TI - Localization of the face area of human sensorimotor cortex by intracranial recording of somatosensory evoked potentials. AB - The authors describe a method of localizing the sensory and motor peri-rolandic cortex representing the face and intraoral structures. Somatosensory evoked potentials (SEP's) to stimulation of the chin, lips, tongue, and palate were recorded in 37 patients studied intraoperatively under general anesthesia or following chronic implantation of cortical surface electrodes. Localization by trigeminal SEP recording was validated by SEP localization of the hand area with median nerve stimulation, and by cortical stimulation of the hand and face areas. The following conclusions were drawn regarding the implementation of face area localization: 1) in general agreement with the results of cortical stimulation in humans and single-unit recordings in monkeys, there is a medial-to-lateral representation in somatosensory cortex of the hand, chin, upper lip, lower lip, tongue, and palate; 2) the chin and lip representations overlap, are adjacent to the hand area, and provide little additional localizing information if the hand area has been identified; 3) stimulation of the tongue and palate evokes reliable, large-amplitude SEP's useful for localization; 4) palatal SEP's allow localization near the sylvian sulcus; 5) for any type of trigeminal stimulation, the largest SEP's are recorded from the somatosensory cortex and provide the most consistent criterion for its identification; and 6) polarity inversion of potentials across the sulcus (a reliable localizing criterion for median nerve SEP's) is a less reliable criterion for trigeminal SEP's. PMID- 8246057 TI - Plasma glucose levels and outcome after aneurysmal subarachnoid hemorrhage. AB - Plasma glucose levels were studied in 616 patients admitted within 72 hours after subarachnoid hemorrhage (SAH). Glucose levels measured at admission showed a statistically significant association with Glasgow Coma Scale scores, Botterell grade, deposition of blood on computerized tomography (CT) scans, and level of consciousness at admission. Elevated glucose levels at admission predicted poor outcome. A good recovery, as assessed by the Glasgow Outcome Scale at 3 months, occurred in 70.2% of patients with normal glucose levels (< or = 120 mg/dl) and in 53.7% of patients with hyperglycemia (> 120 mg/dl) (p = 0.002). The death rates for these two groups were 6.7% and 19.9%, respectively (p = 0.001). The association was still maintained after adjusting for age (> or < or = 50 years) and thickness of clot on CT scans (thin or thick) in the subset of patients who were alert/drowsy at admission. Increased mean glucose levels between Days 3 and 7 also predicted a worse outcome; good recovery was observed in 132 (73.7%) of 179 patients who had normal mean glucose levels (< or = 120 mg/dl) and 160 (49.7%) of 322 who had elevated mean glucose levels (> 120 mg/dl) (p < 0.0001). Death occurred in 6.7% and 20.8% of the two groups, respectively (p < 0.0001). It is concluded that admission plasma glucose levels can serve as an objective prognostic indicator after SAH. Elevated glucose levels during the 1st week after SAH also predict a poor outcome. However, a causal link between hyperglycemia and outcome after delayed cerebral ischemia, although suggested by experimental data, cannot be established on the basis of this study. PMID- 8246058 TI - Dose-response study of the pathological effects of chronically applied direct current stimulation on the normal rat spinal cord. AB - Electrical stimulation of the mammalian central nervous system (CNS) can result in extensive destruction of tissue unless applied within specific stimulation parameters. Classically, unbalanced or monopolar currents have been avoided in order to minimize these harmful effects. However, direct current (DC) fields have recently been proposed for the treatment of spinal cord injury. Until now, no rigorous analysis has been made of the safety of these fields in the mammalian CNS. The purpose of this study was to determine the amount of chronically applied DC current that can be tolerated by the normal rodent spinal cord stimulated with metal disc electrodes. Thirty-five normal rats underwent implantation of DC stimulating devices and were allowed to recover for a period of 2 to 12 weeks. The stimulators delivered constant currents of 0 to 50 microA through two disc shaped platinum/iridium electrodes positioned extradurally at the C-7 and T-3 levels. Following sacrifice of the animals, serial 8-microns cross sections of the spinal cord at the electrode sites were examined microscopically. Evidence of demyelination presumed due to the physical presence of the rostral electrode was seen in animals from most groups including control animals. Pathological changes directly attributable to the applied fields were seen with current as low as 3 microA. It was concluded that DC's of 3 microA or more are harmful to the mammalian CNS with this method of stimulation. In addition, the data suggest that the maximum current density tolerated by the rodent spinal cord is in the order of 75 microA/sq cm. These findings have important implications for the use of chronic DC stimulation in the mammalian CNS. PMID- 8246059 TI - Atlas hypoplasia as a cause of high cervical myelopathy. Case report. AB - A high cervical myelopathy due to atlas hypoplasia is described in a 56-year-old man; the condition caused marked segmental compression of the spinal cord. A remarkable neurological recovery followed decompressive laminectomy of the atlas and adjacent regions. The authors discuss the embryology and etiology of this anomaly. PMID- 8246060 TI - Ganglioglioma presenting as a vascular lesion in a 10-year-old boy. Case report. AB - The authors present the case of a 10-year-old boy admitted for evaluation of a generalized seizure and a history of headaches. Computerized tomography (CT) and gadolinium-enhanced magnetic resonance (MR) imaging demonstrated a large nonhomogeneous contrast-enhancing mass of the left frontal lobe, with a large cystic component. Cerebral angiography revealed the lesion to be highly vascular and fed entirely by the internal carotid artery system. The patient underwent craniotomy and the lesion was completely removed. Neuropathological study revealed that the tumor was a ganglioglioma. On review of the literature, it was found that gangliogliomas often present in the second and third decade, are known to have cystic components, and are contrast-enhancing on CT and MR imaging; however, they are classically known to be avascular on angiography. This case of a markedly vascular ganglioglioma emphasizes that these tumors should be included in the differential diagnosis of vascular supratentorial lesions. PMID- 8246061 TI - Neurogenic hypertension related to basilar impression. Case report. AB - The authors report the resolution of essential hypertension following transoral odontoidectomy and medullary decompression in a 39-year-old woman with basilar invagination. Current understanding of central regulation of the cardiovascular system is discussed and the pertinent neuroanatomy illustrated. Experimental and clinical evidence supporting the role of neurogenic mechanisms in the pathogenesis of hypertension is reviewed. PMID- 8246062 TI - Cranial hemophilic pseudotumor. Case report. AB - Hemophilic pseudotumor is a rare complication of hemophilia occurring in 1% to 2% of individuals with a severe factor VIII or IX deficiency. This pseudotumor has been defined as a progressive cystic swelling involving muscle that is produced by recurrent hemorrhage and may be accompanied by roentgenographic evidence of bone involvement. The case is presented of a 12-month-old child with mild factor VIII deficiency (10% of normal factor VIII activity), who developed a pseudotumor of the skull. PMID- 8246063 TI - Inflammatory pseudotumor of the choroid plexus. Case report. AB - The case of an 18-year-old man with a tumor of the left lateral ventricle is reported. The mass was observed to be contiguous with the choroid plexus but could be totally removed. Pathological examination disclosed an inflammatory pseudotumor. The authors review the literature on this lesion and related disorders involving the central nervous system and stress the need for a morphological differential diagnosis. Inflammatory pseudotumor should be considered in any case of intraventricular tumor. PMID- 8246064 TI - Microvascular sling decompression for trigeminal neuralgia secondary to ectatic vertebrobasilar compression. Case report. AB - A patient with trigeminal neuralgia caused by a tortuous and ectatic vertebrobasilar artery is presented. He was treated with microvascular decompression using a fine silicone sling sutured to the dura over the petrous pyramid. The technical details are described. PMID- 8246065 TI - The Massachusetts General Hospital. Early history and neurosurgery to 1939. AB - The early history of the Massachusetts General Hospital (MGH) is reviewed with emphasis on the development of neurological surgery. The hospital opened in 1823. Early trephinations were performed by Dr. John Collins Warren and others for treatment of trauma and epilepsy. In the 1880's, interest in brain surgery increased, and Dr. John Elliot performed several trephinations for brain tumors, three of which were witnessed by Dr. Harvey Cushing during his years at the MGH as medical student and intern. In 1911, all brain surgery was placed in the hands of Dr. S. J. Mixter. He later shared the assignment with his son, Dr. W. J. Mixter, who described herniation of the intervertebral disc with Dr. J. S. Barr and became the first Chief of the Neurosurgical Service at MGH in 1939. PMID- 8246066 TI - Hearing loss with petrous carotid exposure. PMID- 8246067 TI - Birth trauma and intracranial aneurysm. PMID- 8246068 TI - Aspiration of colloid cyst. PMID- 8246069 TI - Brachytherapy of cystic craniopharyngiomas. PMID- 8246070 TI - Percutaneous endoscopic discectomy. PMID- 8246071 TI - Percutaneous endoscopic discectomy. PMID- 8246072 TI - Facilitators and inhibitors of practice: a faculty perspective. AB - Nurse educators are being encouraged to integrate the role of faculty practice into the role expectations of the educational institution. Schools of nursing are faced with the challenge of facilitating faculty who wish to adopt the practice role. There are limited studies documenting factors that facilitate or inhibit faculty who engage in practice. This article presents the findings of a Delphi procedure used to collect the consensus of practicing faculty concerning organizational and personal facilitators and inhibitors of faculty practice. In this study, 299 faculty representing 170 schools of nursing completed all three rounds of the Delphi. The findings present the top five factors identified by practicing faculty as personal and organizational facilitators and inhibitors of their practice. PMID- 8246073 TI - Development of a predictive model for role strain in registered nurses returning to school. AB - This study identified the individual characteristics that are expected to predict and explain role strain in registered nurses (RNs) returning to school. A model was developed and tested to predict and explain role strain. The characteristics expected to predict role strain were personality, stage of career development, and marital status. Personality was assessed by the Comrey Personality Scales, stage of career development was assessed by the Career Concerns Inventory, and the criterion variable role strain was assessed by the Lengacher Role Strain Inventory. A convenience sample of 123 RN students were asked to participate in the study and 86 RNs volunteered. The data were analyzed using multiple stepwise regression analysis. Results of the stepwise regression analysis identified 13 variables as significant predictors of role strain. A double cross-validation regression procedure was completed to validate the results of the multiple regression analysis. In subsample A of the cross-validation analysis, six variables were found to be significant predictors of role strain, while in subsample B, 12 variables were significant predictors. PMID- 8246074 TI - Accord and discord in students' images of nursing. AB - This article discusses findings from a qualitative study of student nurses' images of nursing from entry to training through early clinical experiences. Three rounds of interviews were conducted with 24 Scottish students. Analysis revealed five major themes in students' initial images: pictures of nursing, the good nurse, what nursing entails, occupational labels for nursing, and being a student/becoming a nurse. These were pursued in later interviews and characteristics of students' experience-mediated images were identified. The picture of adult medical-surgical nursing as typical of real nursing persisted throughout. "Working with people" and "helping" appeared as central characteristics of nursing. Notions of good and bad underlay many features of the image. The paradoxical nature of "involvement" was noticeable. Staff attitudes figured prominently in determining the quality of students' experience and in development of image. Commitment and belonging were integral to students' images and made nursing more than just a job. Processes through which students' images developed included affirmation, accommodation, and nonaccommodation. Associated processes included identification, disillusionment, and extenuation. There was clear evidence of the importance of support to enable students to adapt to image disparities. PMID- 8246075 TI - The effects of imagery abilities and mental rehearsal on learning a nursing skill. AB - Nursing demands competency in numerous psychomotor skills. Using theory from Paivio (1986), Anderson (1976, 1985), and Bandura (1986), this attribute treatment study explored the interactive effects of imagery skills and various combinations of physical and mental practice on learning a skill. The author assessed 108 nursing students for imagery abilities and randomly assigned them to a physical practice (PP), mental rehearsal (MR), or a combined mental rehearsal plus physical practice (MR + PP) group. After learning to apply and remove sterile gloves, subjects were evaluated on their performance. A 3-factor ANOVA was performed and a significant main effect for practice condition was obtained. Subjects in the MR+PP condition performed significantly better than subjects in the MR condition but not the PP condition. These findings have important implications. Incorporating the use of mental rehearsal as an adjunct to physical practice in an effort to facilitate skill acquisition was found to be beneficial. PMID- 8246076 TI - Measuring the nursing, teaching, and interpersonal effectiveness of clinical instructors. PMID- 8246077 TI - Teaching professional writing skills to the baccalaureate student. PMID- 8246078 TI - A five-year evaluation of BSN students in a nursing management preceptorship. PMID- 8246079 TI - Integrating nursing informatics into a graduate research course. PMID- 8246080 TI - A vision for nurse-midwifery education. PMID- 8246081 TI - Women's experiences with combining breast-feeding and employment. AB - A group of primiparas who combined breast-feeding and employment are described in terms of how they felt about the experience, the difficulties they encountered, factors that affected their total duration of breast-feeding, and patterns of feeding while separated from their infant. These women are contrasted with primiparas who weaned prior to returning to work and those who remained homemakers. Of the 619 women included in the analyses, 499 (80.6%) of the women returned to work or school by 12 months postpartum and 288 (46.5%) continued to breast-feed after returning to work. Women who combined breast-feeding and employment were older, had more years of education, worked fewer hours per week, and more worked in professional jobs than those who weaned prior to returning to work. A number of women who chose to express breast milk only at home thought they would have had problems with having enough time and finding a place to express and to store expressed milk had they tried to express breast milk at work. The overwhelming majority of women who combined breast-feeding and employment felt that it was worth the trouble, that they would recommend it to others, and that they had done something special for their infants that no one else could do. The findings suggest that nursing interventions and workplace accommodations could assist more women to experience the benefits and rewards of continuing to breast-feed after returning to employment. PMID- 8246082 TI - Use of a cervical brush for Papanicolaou smear collection. A meta-analysis. AB - Inadequacy of Papanicolaou smear specimens, specifically lack of endocervical cells, has been cited as a major reason why between 0.5% and 44% of cases of dysplasia are not detected by conventional screening tools. Of several sampling instruments devised and tested in an attempt to increase endocervical cell retrieval, one of the most promising in the last decade has been a cervical brush. Studies comparing the cervical brush to more conventional devices such as the spatula and cotton swab were examined in this review. A meta-analysis of these studies indicated a significant mean risk difference or increase in endocervical cell retrieval of 0.22 when the cervical brush was used either alone or in combination with a spatula compared to a spatula alone or with a cotton swab. Subpopulations such as pregnant or postmenopausal women, women with cervical stenosis, or women with a history of cervical surgery also showed significant mean increases in endocervical cell retrieval with the cervical brush. The evidence for whether use of a cervical brush actually increases detection of cervical dysplasia is less clear, although there appears to be a trend in that direction. PMID- 8246083 TI - The Inventory of Functional Status-Fathers. Development and psychometric testing. AB - The Inventory of Functional Status-Fathers (IFS-F) was developed to measure the extent to which men continue or increase their usual household, social and community, childcare, personal care, occupational, and educational activities, and assume infant care responsibilities during their partners' pregnancies and the postpartum period. Content validity was established at 86%. Reliability testing used a sample of 125 expectant fathers and 57 new fathers. Internal consistency reliability using average correlations for the subscale item to subscale total scores ranged from .54 to .75. Subscale to total IFS-F score correlations ranged from .31 to .61. Initial construct validity testing was accomplished by examination of subscale correlations, which ranged from -.02 to .69. The IFS-F may be used to assess the functional status of men during the childbearing period for clinical and research purposes. PMID- 8246084 TI - The ACNM'S visionary planning. PMID- 8246085 TI - Listen to women. The ACNM's vision. AB - This presentation introduced the theme of "Listen to Women" in addressing visionary planning conducted by the American College of Nurse-Midwives in 1993. The author drew on the exemplar nurse-midwifery model of health care created at the Frontier Nursing Service, Leslie County, Kentucky, in 1925 by Mary Breckinridge, RN, MPH, SCM. It is suggested that this historical exemplar remains contemporary to women's health needs today by demonstrating a health system managed by nurse-midwives that provided continuity, accessibility, affordability, accountability, and responsivity to its community through the services and education of nurse-midwives. The author further recommends that the voices of women from the past be threaded to the voices of women in the future in preparation for further visionary planning in addressing women and childbearing family health needs in the 21st century. PMID- 8246086 TI - A challenge for nurse-midwifery. AB - This article suggests that nurse-midwifery today has an exceptional opportunity to play a leadership role in the restructuring of health care policy in the United States. A case is made for the profession to initiate the actions necessary to meet the need for at least 50,000 and possible as many as 100,000 practicing nurse-midwives in this country within the next three decades. The choice for action is in the hands of today's 4,000 nurse-midwives. PMID- 8246087 TI - Nurse-midwifery as it reinforces and transforms the American ideology of gendered roles. AB - Nurse-midwives both reinforce and subvert American constructs of gender roles. They do this both within the context of their relationship with the medical establishment and within the context of their relationship with women and families they serve. The authors explore this paradox through an examination of American society's historical and present constructs of gender role, the ways in which the medical profession has absorbed and prescribed these constructs, the effect of these constructs on medical and nurse-midwifery childbirth management of care, and, within the contextual framework thus established, the role of nurse midwives in reinforcing and subverting traditional gender role constructs. Finally, the impact of this paradox on the provision of nurse-midwifery care to women and families is explored. PMID- 8246088 TI - Effect of meconium on delivery of chorioamniotic membranes. AB - A decrease in stress tolerance of chorioamniotic membranes when exposed to meconium has previously been demonstrated. Clinically, variations in elastic qualities of membranes can be appreciated during their removal following spontaneous vaginal deliveries. The purpose of this study was to determine if there is a relationship between the presence of meconium-stained amniotic fluid and difficulty in removal of chorioamniotic membranes in spontaneous vaginal deliveries. All deliveries over four months (n = 400) were evaluated for color of amniotic fluid and method of delivery of membranes. Overall, the CNMs used significantly more intervention to deliver the membranes if any meconium was present in the amniotic fluid prior to delivery (Fisher's exact test, P = .000004, df = 1). These results suggest that the accoucheur should be prepared for difficulty in membrane removal in deliveries complicated by meconium-stained fluid. PMID- 8246089 TI - Annual student report to the American College of Nurse-Midwives. AB - This article profiles current areas of concern identified by the student membership at the 38th annual American College of Nurse-Midwives convention in June. Communication, financial aid, education, and practice are identified as topics affecting past, present, and future students, as well as the current membership of certified nurse-midwives. Recommendations are offered and the College is petitioned to address these substantial issues that directly impact the membership. PMID- 8246090 TI - Meet Jane Doe. PMID- 8246091 TI - HIV infection. PMID- 8246092 TI - Nursing research and public policy: parental leave. AB - Public policy issues offer fertile ground for the development of research questions. At state and national levels, today's legislative arena bristles with debate on policies affecting the health of women, children, and families. Nurse researchers can contribute to meaningful social debate addressing the benefits and costs of parental leave and other such issues of public policy. This article examines how a relevant social issue can help nurses identify research questions important to the development of sound public policy. PMID- 8246093 TI - Litigation, electronic fetal monitoring, and the obstetric nurse. AB - Using excerpts from actual lawsuits that allege malpractice and name obstetric nurses as defendants, this article explores the process and some possible outcomes of malpractice litigation. It discusses the duty of the nurse, the concept of reasonably prudent practice, and the role of the expert. In addition, it identifies some institutional standards that can mitigate potential damages in malpractice claims. PMID- 8246094 TI - Bed rest during pregnancy: implications for nursing. AB - Research on nonpregnant individuals reveals that bed rest produces numerous adverse physical and psychosocial effects. Evidence shows that bed rest affects pregnant women in the same manner. This article provides an overview of the history of bed rest, research on the side effects of bed rest, and a discussion of the research that has been conducted on these side effects during pregnancy. Also discussed are implications for providing antepartum, intrapartum, and postpartum nursing care. PMID- 8246095 TI - Adverse exposures and use of universal precautions among perinatal nurses. AB - OBJECTIVE: To identify the level of protective barrier use by perinatal nurses, the numbers of exposures experienced, and the factors influencing barrier use. DESIGN: Descriptive correlational study. PARTICIPANTS: A random sample of 560 perinatal nurses practicing in Arizona were sent questionnaires; 306 nurses responded. MAIN OUTCOME MEASURES: The nurses indicated frequency of barrier use, on a Likert-type scale, for 15 common obstetric procedures involving risk for blood or body fluid exposure. The nurses also reported number of exposures and barrier failures experienced in the preceding year. Two other scales were used to identify factors influencing barrier use and nonuse. RESULTS: A low frequency of barrier use and a high frequency of exposures were reported. A relationship was found between barrier use and exposures (r = -.29, p < .0001): As barrier use increased, exposures to body fluids decreased somewhat. Suspicion that a patient was a member of a high-risk group (e.g., an intravenous drug user) was a strong motivator for barrier use. Poorly fitting gloves and the need for quick action were strong influences for barrier nonuse. CONCLUSIONS: Perinatal nurses were inadequately protected from bloodborne disease. PMID- 8246096 TI - Sibling behavior with a newborn: parents' expectations and observations. AB - OBJECTIVE: To investigate parents' expectations and perceptions of their firstborn child's behavior before and after the birth of a second child. DESIGN: Couples expecting their second child completed questionnaires before and after the second child's birth. SETTING: Two large private hospitals offering primary care in urban settings. PARTICIPANTS: Seventy married couples who were expecting their second child and had one child age 6 or younger at home. MAIN OUTCOME MEASURES: The Older Child Expectations, Older Child Observations, and Sibling Behavior inventories. STATISTICAL ANALYSIS: Independent t-tests were used to measure mean changes over time. RESULTS: For the items related to the newborn on the Older Child Expectations and Observations inventories, 16 of 23 items differed significantly over time for one or both parents. Parents expected their firstborn child's behavior to be more negative than they actually observed it to be. For the items dealing with developmentally appropriate behaviors, only 7 of 45 items differed significantly over time. CONCLUSIONS: Parents expected more negative behaviors in their firstborn child than they actually observed after the birth of their second child. Parents observed similar levels of developmentally appropriate behaviors in their firstborn child both before and after the birth of the second child. PMID- 8246097 TI - Breastfeeding care in Ohio hospitals: a gap between research and practice. AB - OBJECTIVE: To assess current breastfeeding care in Ohio hospitals and compare that care to research-based principles. DESIGN: Survey. SETTING: Ohio hospitals that provide maternity care. PARTICIPANTS: All 141 Ohio hospitals that provide maternity care were invited to participate. One-hundred sixteen (83%) hospitals returned usable surveys completed by obstetric nurse managers. MAIN OUTCOME MEASURE: A 38-item questionnaire provided data on hospital demographics and information regarding the care of breastfeeding mother-infant dyads. RESULTS: Research-based practices common in Ohio's hospitals include demand feeding, breastfeeding education, and breastfeeding as the initial neonatal feeding. Common non-research-based practices include supplemental fluid administration, postpartum nipple treatments, mandatory initial nursery stays, limited sucking time, restricted maternal-infant contact, distribution of formula packs, minimal follow-up care, and the suspension of breastfeeding for hyperbilirubinemia. CONCLUSIONS: Despite positive changes in perinatal care, a number of non-research based practices persist in Ohio hospitals for the care of the breastfeeding mother-infant dyad. PMID- 8246098 TI - Health, symptoms, self-care, and dyadic adjustment in menopausal women. AB - OBJECTIVE: To determine the relationships among perceived health, menopausal symptoms, and self-care responses in perimenopausal and postmenopausal women. DESIGN: Descriptive, cross-sectional survey. SETTING: A large midwestern city in the United States. PARTICIPANTS: One-hundred-one menopausal women who were over age 40, used no hormones during the previous 6 months, and had an intact uterus. MAIN OUTCOME MEASURES: Four standardized survey instruments. RESULTS: Significant correlations were found between self-rated health and health perceptions (r = .44, p = .0004), self-rated health and total symptoms (r = -.30, p = .0023), self rated health and worrisome symptoms (r = -.26, p = .0085), health perceptions and worrisome symptoms (r = .30, p = .0195), health perceptions and self-care responses (r = .43, p = .0009), and total symptoms and worrisome symptoms (r = .38, p = .0001). CONCLUSIONS: Health is related to menopausal symptoms and self care. Dyadic adjustment is not related to health, menopausal symptoms, or self care. Healthy menopausal women provide self-care and need not automatically be treated for menopause. PMID- 8246099 TI - Poor, poorer, poorest. PMID- 8246100 TI - Laparoscopic assisted vaginal hysterectomy: a report of 26 cases. AB - This study was undertaken to evaluate the feasibility and safety of laparoscopic assisted vaginal hysterectomy. We have retrospectively studied the first twenty six cases. All cases were reviewed and included in this report. The complication rate is very minimal and well below that which would be expected from abdominal or vaginal hysterectomy approach. The operative time is approximately double that of a standard approach; however, the hospital stay is greatly decreased and the patient's return to normal activity has been considerably shortened when compared to the standard abdominal procedure. We conclude that the operation is not only feasible, but should be considered any time an abdominal approach to hysterectomy is being contemplated. PMID- 8246101 TI - Health care tips for your professional corporation or where there is a buy-sell agreement, there is a way. PMID- 8246102 TI - Guardianship in Oklahoma for purposes of medical consent. PMID- 8246103 TI - Diabetic retinopathy ... we can make a difference! PMID- 8246104 TI - General counsel defends state medical licensure board. PMID- 8246105 TI - Appraisals of childbirth experience and newborn characteristics: the role of hardiness and affect. PMID- 8246106 TI - Self-identity and specific vulnerability to depressed mood. AB - Four studies were conducted to test the hypothesis that experiencing a life event that disrupts the pursuit of self-defining activities would be associated with depressed mood. Across all four studies, a strong and exclusive identification with the athlete role was related to subjects' affective response to both hypothetical and actual athletic injuries. The results highlight the importance of assessing the match between specific cognitive diatheses and specific life events in predicting depressive reactions. PMID- 8246107 TI - Dependent and self-critical depressive experiences among inner-city adolescents. AB - The Adolescent Depressive Experiences Questionnaire was administered to 142 inner city ninth graders. Based on theory, several potential correlates of dependency and self-criticism were selected for study, including depression, aspects of anxiety, locus of control, and facets of social behavior. Self-criticism showed stronger associations with depression and concerns about losing others' approval, while dependency showed stronger associations with diffuse tension and worry and sensitivity to others' behavior. The two depressive tendencies had comparable associations with physiological anxiety and external locus of control. These findings are discussed in terms of current perspectives on the symptomatic heterogeneity of depressive experiences. PMID- 8246108 TI - Beyond personality impressions: effects of physical and vocal attractiveness on false consensus, social comparison, affiliation, and assumed and perceived similarity. AB - We examined the effects of target persons' physical and vocal attractiveness on judges' responses to five measures: false consensus (the belief that the target shares one's behavior), choice of targets as comparison others, affiliation with targets, assumed similarity (similarity between self-ratings and ratings assigned to targets), and perceived similarity (direct questions about similarity). Higher physical attractiveness and higher vocal attractiveness were both related to higher scores on all variables. The effect of one type of attractiveness was more pronounced for higher levels of the other type of attractiveness. The joint effect of the two types of attractiveness was best described as synergistic, i.e., only targets high on both types of attractiveness elicited higher scores on the dependent variables. The effect of physical attractiveness on most dependent variables was more pronounced for subjects who were themselves physically attractive. The synergistic effect (the advantage of targets high on both types of attractiveness) was more pronounced for judges high in self-monitoring. The contribution of the study to the literature on attractiveness stereotypes is discussed. PMID- 8246109 TI - Sex differences in intensity of emotional experience: a social role interpretation. AB - According to gender role theory, women's greater emotional intensity than men's stems from normative expectations for sex differences that arise as a result of men's and women's social roles. In the 1st experiment, endorsement of normative expectations for sex differences was associated with sex differences in Ss' own emotions: To the extent that they endorsed stereotypical differences between men and women, female Ss reported personally experiencing emotions of greater intensity and male Ss reported experiencing emotions of lesser intensity. The 2nd study manipulated expectations for responsiveness while Ss viewed a series of emotion-inducing slides. When instructions rendered normative expectations comparable for men and women, no sex differences were obtained in emotion self reports. Furthermore, women evidenced more extreme electromyograph physiological responding than men, suggesting general sex differences in emotion that are not limited to self-report. PMID- 8246110 TI - The role of situational demands and cognitive competencies in behavior organization and personality coherence. AB - Consistency in the natural organization of aggressive and prosocial (constructive) behavior, assessed repeatedly in vivo over a summer in a residential camp for children, was predicted from situational and personal characteristics. Similarity of situations in the types of competencies they demand in part predicted cross-situational consistency in individual differences in aggressive behaviors (Study 1). Study 2 examined the effect of cognitive competence on the discriminative patterning of behavior variation across situations. More cognitively competent Ss showed such discriminative patterning, which was reflected in greater Person X Situation interaction variance in their prosocial behavior. PMID- 8246111 TI - Authoritarianism and sexual aggression. AB - In Study 1, 198 men completed the Right Wing Authoritarianism, Sex Role Ideology, Hostility Towards Women, Acceptance of Interpersonal Violence, Adversarial Sexual Beliefs, and Rape Myth Acceptance scales, as well as measures of past sexually aggressive behavior and likelihood of future sexual aggression. As predicted, authoritarianism and sex role ideology were as closely related to self-reported past and potential future sexually aggressive behavior as were the specifically sexual and aggression-related predictors. Among 134 men in Study 2, authoritarianism and sex guilt positively correlated with each other and with self-reported past sexual aggression. In both studies, the relationship of authoritarianism and sexual aggression was larger in community than in university samples. PMID- 8246112 TI - Extraversion and neuroticism as predictors of objective life events: a longitudinal analysis. PMID- 8246113 TI - Life stress and dysphoria: the role of self-esteem and hopelessness. AB - This study evaluated the hypotheses that the relation between life stress (life events and daily hassles) and longitudinal change in dysphoria would be (a) moderated by self-esteem and (b) mediated by longitudinal change in hopelessness. Eighty undergraduates were first assessed on self-esteem, hopelessness, and dysphoria and then reassessed 3 months later on life events, daily hassles, hopelessness, and dysphoria. Residual change in dysphoria was significantly associated with self-esteem, life stress, and a Self-Esteem X Life Stress interaction. However, inconsonant with predictions, the moderating impact of self esteem was greatest under conditions of low (vs. high) life stress. Moreover, residual change in hopelessness mediated the relations between residual change in dysphoria and both self-esteem and life stress. PMID- 8246114 TI - Motivated resistance and openness to persuasion in the presence or absence of prior information. AB - Three experiments investigated the relation between need for cognitive closure and persuasion. In the 1st study, Ss high on an individual-differences measure of need for closure were more resistant to persuasion by their low need-for-closure counterparts than vice versa. In the 2nd study, Ss in a noisy environment, assumed to instill a relatively high need for closure, were more resistant to persuasion than Ss in a quiet environment, but only in presence of an initial informational base for an opinion. In its absence, Ss in the noisy (vs. quiet) environment were less resistant to persuasion. The interaction between need for closure and informational base was replicated in the 3rd experiment reverting to the individual-differences measure of need for closure. The discussion considered implications of these findings for further persuasion phenomena. PMID- 8246115 TI - In search of the "hot" cognitions: attributions, appraisals, and their relation to emotion. AB - Two studies examined the hypothesized status of appraisals, relative to attributions, as proximal antecedents of emotion. In Study 1, which looked at 6 emotions (happiness, hope-challenge, anger, guilt, fear-anxiety, and sadness), undergraduates (N = 136) reported on their attributions, appraisals, and emotions during past encounters associated with a variety of situations. In Study 2, which was focused on anger and guilt, undergraduates (N = 120) reported on these same variables in response to experimenter-supplied vignettes that systematically manipulated theoretically relevant attributions. The results of both studies indicated that the emotions were more directly related to appraisals than they were to attributions, and Study 2 provided evidence that appraisal serves as a mediator between attribution and emotional response. These findings lend support to the hypothesized status of appraisal as the most proximal cognitive antecedent of emotion. PMID- 8246116 TI - Love, hate, anger, and jealousy in close relationships: a prototype and cognitive appraisal analysis. AB - The aim of this research was to make a prototype and cognitive appraisal analysis of 4 emotions within marriage. In Study 1, 160 Ss recalled and wrote about a partner-related love, hate, anger, or jealousy incident. Distinct prototypes and appraisal patterns were obtained. In Study 2, 80 Ss wrote accounts of hypothetical love, hate, anger, and jealousy events in marriage. The results suggested both recalled and hypothetical accounts were derived from the same knowledge structures. In Study 3, Ss matched emotions to events described with varying amounts of prototypical and appraisal information, derived from Study 1. Adding such information significantly increased emotion-matching accuracy over the event-description-only condition. The results are discussed in relation to prototype and cognitive appraisal theories of emotion in close relationships. PMID- 8246117 TI - Melanized poly(HEMA) hydrogels: basic research and potential use. AB - Synthetic melanogenesis, using epinephrine and other melanin precursors, within the matrix of hydrophilic polymers and copolymers of 2-hydroxyethyl methacrylate resulted in hydrogels able to absorb ultraviolet and visible radiation. This significantly enhances their value as materials for extraocular (contact lenses) or intraocular (artificial crystalline lenses) devices that should protect the retina of aphakic patients from potential damage induced by light. The two-phase morphology of melanized hydrogels, as investigated by TEM, revealed a fine structure that is possibly indicative of a true sequential interpenetrating polymer network. Their biocompatibility was evaluated by a set of different assays involving human choroidal fibroblasts. No cytotoxicity was found in the aqueous extracts of materials. By using an assay with cells and polymers embedded in a collagen gel, a short-range toxic effect was detected, presumably caused by melanin itself. However, in vivo experiments in animal eyes with melanized hydrogel intraocular lenses did not reveal any toxic reaction. PMID- 8246118 TI - Hydrogen grafted polymer surfaces: interaction and morphology of platelets. AB - Besides the quantification of platelet adhesion, the morphology of adherent platelets on a polymer surface seems to be important as far as thrombogenicity is concerned. It is suggested that such events usually take place on a layer of adsorbed proteins on the surface of the blood. In the present study, we observed the morphological variation of adherent calf platelets (in vitro under static conditions) on bare and hydrogel-grafted Angioflex sheets. Electron micrographs indicate the extent of deformation of the Angioflex sheets varying with modified substrates as compared to the bare sheets. PMID- 8246119 TI - Controlling the risks of a medically complicated pregnancy. PMID- 8246120 TI - Chronic hypertension in pregnancy: focus on clinical management. PMID- 8246121 TI - Pregnancy in women with thyroid disease: a delicate balance. PMID- 8246122 TI - Effects of systemic lupus erythematosus on pregnancy and the neonate. PMID- 8246123 TI - Neonatal nurse practitioners: paving the way for case management of chronically ill infants and their families. PMID- 8246124 TI - Congestive heart failure: pathophysiology and management throughout infancy. PMID- 8246125 TI - Chronic psychiatric illness and pregnancy: nursing implications. PMID- 8246126 TI - Temporal characteristics of non-oral feedings and chronic feeding problems in premature infants. PMID- 8246127 TI - Serotonin-induced muscle contraction in rat stomach fundus is mediated by a G alpha z-like guanine nucleotide binding protein. AB - Serotonin (5-HT) potently contracts the fundus of the rat stomach; however, the associated transduction pathway has not been described fully. Experiments were performed in an attempt to gain insight into the coupling mechanism associated with this fundal 5-HT receptor. 5-HT-stimulated [35S]GTP gamma S binding to a protein which was recognized by anti-G alpha Z antiserum in a Mg(++)-dependent fashion. 5-HT increased [35S]GTP gamma S binding in the fundus, but not in the corpus of the rat stomach. 5-HT also enhanced the binding of [alpha-32P]GTP to the fundal protein and increased the hydrolysis of GTP to GDP in fundal membranes. The fundal protein which binds GTP is 25 to 29 kDa in size whereas the brain G alpha Z protein which is recognized by the anti-G alpha Z antibody is a 41 kDa protein. Mixing experiments revealed that the fundal guanine nucleotide binding protein does not appear to be a proteolytic product of the 41 kDa G alpha Z protein. Activating protein kinase C with phorbol-12-myristate, 13-acetate induced a concentration-dependent, noncompetitive inhibition of [35S]GTP gamma S binding to the fundal protein, and of 5-HT-induced contraction of fundal strips. Phorbol-12-myristate, 13-acetate did not alter carbachol- or KCl-mediated fundus contraction. Furthermore, the activation of [35S]GTP gamma S binding by serotonergic agonists and its inhibition by pharmacological antagonists corresponded to the known actions of these agents on contraction of fundal muscle. The results provide evidence that the 5-HT receptor in the rat stomach fundus is coupled directly or indirectly to a G alpha z-like protein which may mediate 5-HT-induced contraction in this tissue. PMID- 8246128 TI - Identification of CYP2C23 expressed in rat kidney as an arachidonic acid epoxygenase. AB - A cDNA was isolated from a rat kidney cDNA library using mixed probes of CYP (cytochrome P450) 2C6, 2C7, 2C8, 2C9 and 2C18 cDNAs. The 3'-terminal and 5' terminal regions of the cDNA were sequenced and were identical with those of cytochrome P450 2C23 (CYP2C23) except for a one-base deletion and a one-base addition in coding region. These changes caused a frame shift and changed the deduced amino acid sequence relative to the previously published sequence. This cDNA was expressed using a baculovirus expression system, and the resultant P450 had a lambda max of 450 nm when reduced and complexed with carbon monoxide. Specific content of the expressed P450 ranged from 0.27 to 0.43 nmol/mg of cell lysate protein. Arachidonic acid metabolism catalyzed by expressed CYP2C23 indicated that CYP2C23 efficiently produced epoxyeicosatrienoic acids (EETs). These EETs were characterized further by gas-liquid chromatography/negative ion chemical ionization mass spectrometry (GC/NCIMS) and were found to include 8,9 EET, 11,12-EET and 14,15-EET in a ratio of 1:2:1. No 5,6-EET was detected. A low rate of lauric acid hydroxylation at the (omega-1)-position was found, but the enzyme was unable to metabolize prostaglandin E1. These studies suggest that CYP2C23 is responsible, in part, for the production of EETs in rat kidney. PMID- 8246129 TI - Angiotensin II-induced vasopressin release is mediated through alpha-1 adrenoceptors and angiotensin II AT1 receptors in the supraoptic nucleus. AB - The central actions of angiotensin II (ANG II) include the release of vasopressin (AVP) from the supraoptic nucleus (SON) via the pituitary gland into the blood. In conscious rats, we investigated whether catecholamines in the SON are involved in this release process. It was found that i.c.v. injections of ANG II (100 ng) selectively increased the release of norepinephrine (NA) from the SON. Like the ANG II i.c.v.-induced AVP release, this effect was prevented by i.c.v. pretreatment with the ANG II AT1 receptor antagonist, losartan (5 micrograms). The alpha-1 adrenoceptor antagonist, prazosin (0.7 nmol), injected bilaterally into the SON, significantly reduced the ANG II 100-ng i.c.v.-induced AVP release. Pretreatment with the alpha-2, beta-1 and beta-2 adrenoceptor antagonists, idazoxan, atenolol and ICI 118551, respectively, had no effect. Injections of NA into the SON increased plasma AVP at doses up to 10 nmol but not at higher doses (30-100 nmol). The effects of NA were mimicked by the alpha-1 adrenoceptor agonist, methoxamine (1-5 nmol). Bilateral pretreatment of the SON with losartan (5 micrograms) markedly inhibited the i.c.v. ANG II 100 ng-induced AVP release. The increase in AVP release after ANG II injections into the SON was also inhibited by losartan pretreatment into the SON, whereas prazosin had no effect. These results demonstrate that the ANG II-induced release of AVP is initiated through periventricular ANG II AT1 receptors and involves postsynaptic alpha-1 adrenoceptor stimulation in the SON. In addition, ANG II AT1 receptors in the SON can contribute to AVP release after periventricular ANG II receptor stimulation. PMID- 8246130 TI - Mechanism of flecainide's rate-dependent actions on action potential duration in canine atrial tissue. AB - Increases in action potential duration (APD) caused by most antiarrhythmic drugs are maximal at slow rates and are attenuated during tachycardia, causing decreased action during arrhythmias and maximum effects during sinus rhythm. This property, "reverse use-dependence," limits efficacy and contributes to proarrhythmic potential. We have shown that the class 1c antiarrhythmic drug flecainide increases atrial APD to a greater extent at faster rates and that this property may underlie some of the drug's antiarrhythmic actions. The present studies were designed to evaluate possible underlying ionic mechanisms. Standard whole-cell voltage clamp and microelectrode techniques were used to study ionic currents and action potentials of canine atrial tissue. Flecainide (4.5 microM) increased APD at cycle lengths ranging from 150 to 1000 msec and attenuated the APD shortening that resulted from increased activation rate, resulting in greater APD prolongation at faster rates. The major time-dependent outward current (Ito), was reduced by flecainide in a rate-independent fashion. Flecainide's effect on Ito was due to inhibition of the 4-aminopyridine-sensitive component (Ito1); flecainide did not alter inward calcium current or the calcium-sensitive component of Ito (Ito2). The specific sodium channel blocker tetrodotoxin (1 microM) and the Na+, K(+)-ATP'ase inhibitor ouabain (1 microM) suppressed rate dependent APD shortening in a fashion similar to flecainide, and both flecainide and ouabain attenuated postoverdrive membrane hyperpolarization. We conclude that the rate-dependence of flecainide's action on APD is not explained by use dependent changes in outward currents but may be due to sodium channel blockade resulting in decreased sodium loading and reduced Na+, K(+)-ATP'ase stimulation during tachycardia. PMID- 8246131 TI - 3-Deazaadenosine: a therapeutic strategy for myasthenia gravis by decreasing the endocytosis of acetylcholine receptors. AB - The basic abnormality in myasthenia gravis is the depletion of acetylcholine receptors (AChRs) at neuromuscular junctions, which is due in part to excessive endocytosis brought about by the action of pathogenic antibodies. We asked whether 3-deazaadenosine (3DZA), an inhibitor of phospholipid methylation, could decrease the rate of endocytosis of muscle AChRs and thereby interfere with this pathological process. The rationale for the use of 3DZA is that methylation of phospholipids alters membrane properties, and inhibition of methyltransferase reactions is known to slow the process of endocytosis. In this study, we have tested the effects of 3DZA and other methylation inhibitors on the degradation and synthesis of AChRs in an in vitro model of myasthenia gravis, using primary rat skeletal muscle cultures and serum from human myasthenic patients. In normal cultures (without myasthenic serum), 3DZA inhibited AChR degradation with a broad dose-response relationship, beginning as low as 2 microM (P < .0001). There was no acute effect on synthesis of AChRs or on other measures of muscle cell integrity. When human myasthenic serum was added to the cultures to accelerate the endocytosis and degradation of AChRs, 3DZA still potently inhibited the degradation rate. Because the drug allows accumulation of AChRs in the surface membrane of the muscle cell by reducing endocytotic degradation, it provides a potential strategy for therapy in human myasthenia gravis. PMID- 8246132 TI - Characterization of the tachykinin neurokinin-2 receptor in the human urinary bladder by means of selective receptor antagonists and peptidase inhibitors. AB - The tachykinin (NK2) receptor-mediating contraction of the human isolated bladder to NKA was investigated by studying the affinities of eight structurally different receptor-selective antagonists (linear peptides, cyclic peptides and pseudopeptides, nonpeptide NK2 receptor antagonists). The affinities of the antagonists were compared to those measured for the same ligands at the NK2 receptors previously characterized in the rabbit pulmonary artery and hamster trachea. In the presence of a cocktail of peptidase inhibitors (bestatin captopril and thiorphan, 1 microM each) no significant correlation was found between pA2 values measured in the human bladder vs. those measured in the other two NK2 receptor-bearing preparation. In the presence of the aminopeptidase inhibitor amastatin, however, pA2 values of linear antagonists bearing an N terminal Asp residue MEN 10,207 and MEN 10,376 were significantly enhanced and these pA2 values were used for analysis; a significant correlation was found between pA2 values measured in the human urinary bladder and rabbit pulmonary artery. The pseudopeptide analog of NKA (4-10), MDL 28,564 which also bears a N terminal Asp residue behaved as an agonist and its action was enhanced by amastatin. We conclude that the NK2 receptor-mediating contraction of the human urinary bladder smooth muscle is similar to that previously characterized in the rabbit pulmonary artery (NK2A receptor category); in the human bladder smooth muscle an amastatin-sensitive peptidase (possibly aminopeptidase A) limits biological activity of linear peptide derivatives of NKA bearing a N-terminal Asp residue. PMID- 8246133 TI - Bronchopulmonary actions of 1-(1,2,3,4-tetrahydro-1-naphthylenyl)-1H-imidazole, nitric acid salt (LY150310), a substituted imidazole, in the guinea pig. AB - 1-(1,2,3,4-tetrahydro-1-naphthylenyl)-1H-imidazole, nitric acid salt (LY150310), was examined for bronchodilator activity in the guinea pig. In guinea pig tracheal preparations, LY150310 competitively antagonized the contractile effects of exogenous histamine and blocked the histamine-mediated component of contractions produced by ovalbumin challenge. LY150310 had little effect on the nonhistamine component of ovalbumin-induced contractions of lung parenchymal strips, but it enhanced the production of prostaglandin (PG) E2 and PGF2 alpha although it partially inhibited thromboxane B2 formation. In other studies, in which postmortem pulmonary gas trapping was used as an index of in vivo airway obstruction, i.v. LY150310 dose-dependently inhibited the bronchospasm produced by aerosols of the divalent cationic ionophore A23187, histamine, 5 hydroxytryptamine, leukotriene D4, methacholine, ovalbumin or platelet activating factor. LY150310 was equal to or more potent than aminophylline in all test systems. Also, orally administered LY150310 inhibited the airway obstruction produced by selected challenge aerosols. In ex vivo studies, LY150310 elevated PGE2 and tended to decrease thromboxane B2 in sodium arachidonate-stimulated whole blood. However, PGE2 and other cyclooxygenase products did not appear to account for in vivo bronchodilation, because combining LY150310 and piroxicam did not alter inhibition of A23187-induced airway obstruction. Our results demonstrate that LY150310 reduces airway obstruction caused by a variety of bronchoconstrictive agents, including A23187 and ovalbumin. Although this substituted imidazole appears to have activity as a histamine H1-receptor antagonist and can alter prostanoid concentrations in vitro and in vivo, its mode of bronchodilation is unclear. PMID- 8246134 TI - Aminopyridine block of potassium channels in mouse neuroblastoma cells. AB - Although 4-aminopyridine (4-AP) is known to block a variety of voltage-dependent K channels, details as to the site of action and the mechanism of block are known for relatively few. Single channel analysis has not been extensively used to answer these questions. The actions of 4-AP on whole cell K currents and single voltage-dependent K channels that exhibit fast activation and inactivation were therefore examined in N1E-115 neuroblastoma cells. The concentration for half block (K0.5) of the whole cell K current for externally applied compounds was found to be 56 microns for 4-AP and 0.3 mM for 3,4-diaminopyridine. 4-AP slowed the rate of development of outward K current, and the rate of decay after repolarization. These effects were consistent with the idea that 4-AP preferentially blocked a type of K channel generating a transient current. Block of this component of current was time- and use-dependent. 4-AP blocked the channel responsible for the transient outward current by decreasing the probability of an open channel in inside-out patches. 4-AP reduced the open time, indicating that 4-AP can interact with the open channel. The first latency to opening was also increased. 4-Aminopyridine methiodide (4-APMI), a permanently charged derivative, blocked the whole cell current with a K0.5 = 0.19 mM. Block by 4-APMI was found to be by a different mechanism at a different site compared to 4-AP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246135 TI - Calcium channel blockers inhibit hydrogen peroxide-induced proliferation of cultured rat mesangial cells. AB - The present experiments were designed to assess a possible role of H2O2 in the proliferation of cultured rat mesangial cells, as well as to evaluate the effect of different calcium channel blockers and a platelet-activating factor antagonist on this proliferation. Cultured rat mesangial cells were plated at two densities (10,000 and 25,000 cells/well) in 24-well dishes, and proliferation was measured by analyzing [3H]thymidine incorporation and by directly counting the cells. Hydrogen peroxide, 100 microM, increased [3H]thymidine incorporation at the two densities tested (85 and 59%, respectively), as well as the number of cells (53 and 23%, respectively). This effect was dose dependent and it was blocked completely by verapamil and diltiazem, 10 microM, but not by TMB-8 [3,4,5 trimethoxybenzoic acid 8-(diethylamino) octyl ester] at the same concentration. BN 52021, a competitive antagonist of platelet-activating factor, only slightly blocked the H2O2-dependent proliferation. The inhibitory action of the two calcium antagonists tested started at concentrations as low as 1 nM, and inhibited completely the H2O2 stimulated proliferation at concentrations between 0.1 and 1 microM. These results establish that H2O2 is able to induce proliferation of mesangial cells. Although the pathophysiological implications of this finding remain to be proven, these data suggest a potential therapeutic action of calcium antagonist in inflammatory conditions such as glomerulonephritis. PMID- 8246136 TI - Tamoxifen and vanadate synergize in causing accumulation of polyphosphoinositides in GH4C1 membranes. AB - Tamoxifen caused a dramatic stimulation of phosphatidylinositol kinase and phosphatidylinositol-phosphate (PIP) kinase activity in GH4C1 membrane preparation with an ED50 of 20 microM. Vanadate ions alone did not appreciably elevate the amount of polyphosphoinositides; however, when added together with tamoxifen it synergistically enhanced the formation of PIP and phosphatidylinositol-bisphosphate (PIP2). Vanadate caused the inhibition of phosphomonoesterase activity in the membranes that converts PIP2 to PIP and PIP to phosphatidylinositol. The synergism between tamoxifen and vanadate thus results from tamoxifen-induced stimulation of the phosphoinositide kinase reaction and vanadate inhibition of the backward phosphomonoesterase reaction. Tamoxifen had no effect on phosphomonoesterase activity. With optimal concentrations of the drugs, PIP was increased from 8.3 to 75%, and PIP2 was augmented from 0.36 to 8.5% of the total membrane phosphoinositides. Tamoxifen and vanadate are thus useful tools for the investigation of phosphoinositides metabolism. PMID- 8246137 TI - RWJ 26629, a new potassium channel opener and vascular smooth muscle relaxant: a potential antihypertensive and antianginal agent. AB - The effects of trans-5,6-dihydro-6-hydroxy-5,5-dimethyl-2-nitro-7-(2-oxopiperidin -1-yl)-7H- thieno[3,2-b]pyran (RWJ 26629) were compared with those of the standard potassium channel opener cromakalim and several standard calcium channel blockers. RWJ 26629 lowered the mean arterial blood pressure in conscious spontaneously hypertensive (ED30 = 10 micrograms/kg p.o. or 8 micrograms/kg i.v.) and renal hypertensive (15 micrograms/kg p.o.) rats, conscious renal hypertensive (ED20 = 4 micrograms/kg p.o.) and normotensive (ED20 = 5 micrograms/kg p.o. or 2 micrograms/kg i.v.) dogs and anesthetized rhesus monkeys (ED20 = 6 micrograms/kg i.v.). RWJ 26629 was more potent than cromakalim and had a maximal activity greater than the calcium channel blockers. At antihypertensive doses, RWJ 26629 had no significant effect on cardiac force, cardiac output, stroke volume or stroke work in dogs and had little or no effect on renal, carotid or femoral blood flow or vascular resistance. RWJ 26629 was also selective for antihypertensive activity in rats compared with its ability to inhibit intestinal motility. However, RWJ 26629 did relax contracted pulmonary smooth muscle in vivo at antihypertensive doses. All compounds tested caused reflex tachycardia in conscious dogs, although this effect was lowest for RWJ 26629. Most importantly, RWJ 26629 potently and selectively increased coronary blood flow with a potency and duration of action greater than that of cromakalim or nifedipine without affecting contractile force. In vitro, RWJ 26629 selectively relaxed precontracted coronary arteries compared with its effect on femoral arteries.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246138 TI - Angiotensin II receptor blockade improves renal function in rats with reduced renal mass. AB - The effect of inhibiting the renin-angiotensin system was evaluated in male Sprague-Dawley rats with reduced renal mass produced by right nephrectomy and infarction of two-thirds of the left kidney. Separate groups of rats were then administered the angiotensin receptor antagonists, A-81988 or losartan (DuP 753), the angiotensin converting enzyme inhibitor, enalapril, or vehicle (tap water) in their drinking water for 4 weeks. Tail cuff blood pressures and blood samples were obtained weekly. Excretory function during week 4 was evaluated using metabolic cages. Rats with reduced renal mass were characterized by a significant elevation in systolic blood pressure and urinary protein excretion along with a reduced urine osmolality. At 1 mg/kg/day, A-81988 prevented the hypertension and the development of proteinuria. A-81988 administration also improved urinary concentrating ability because urine osmolality was significantly higher in this group compared to untreated controls. The same dose of losartan or enalapril was ineffective at controlling the development of the hypertension, indicating that A 81988 is more potent in vivo. Despite the maintenance of systemic hypertension, losartan significantly blunted the proteinuria compared to vehicle-treated controls. At a dose of 10 mg/kg/day, losartan and enalapril also prevented the increase in systolic blood pressure and proteinuria and produced an increase in urine osmolality. These data support the hypothesis that angiotensin receptor antagonists have beneficial effects in forms of renal failure associated with proteinuria and diminished concentrating ability. PMID- 8246139 TI - NPC 15669 reduces mortality associated with sepsis in rats. AB - N-[9H-(2,7-dimethylfluoren-9-ylmethoxy)carbonyl]-L-leucine (NPC 15669), a leukocyte recruitment inhibitor, was investigated for its ability to enhance survival in a rat model of sepsis, fecal peritonitis. Infusion of NPC 15669 (3 mg kg-1 hr-1 i.v.) for 19, 24 or 48 hr or three to four bolus injections (10 mg/kg) at 2- or 6-hr intervals along with gentamicin effectively cured all animals (> 2 week survival) relative to gentamicin-treated controls (28 +/- 1 hr survival), whereas infusion at a 10-fold lower dose was ineffective. Unlike aspirin and dexamethasone, which were inactive (< 36-hr survival), ibuprofen significantly increased the survival time (66 +/- 1 hr) but did not cure septic rats. The efficacy of NPC 15669 on survival was associated with the reversal of leukopenia and a marked inhibition of neutrophil infiltration into the small intestine. By contrast, i.v. bolus injection or infusion of a related analog, N-[9H-fluoren-9 ylmethoxy)carbonyl]glycine, which does not inhibit leukocyte recruitment, failed to reduce the mortality rate associated with fecal peritonitis-induced sepsis. In addition, NPC 15669 was efficacious therapeutically, even when administered as late as 6 hr after the induction of sepsis (14 of 16 animals survived > 5 days). Together, these data suggest that NPC 15669 may be useful in the treatment of septic shock. PMID- 8246140 TI - Role of calcium in plasma protein binding and renal handling of alendronate in hypo- and hypercalcemic rats. AB - Alendronate (4-amino-1-hydroxybutylidine-1,1-bisphosphonate), an antiosteolytic agent, is currently under investigation for the treatment of osteoporosis. Earlier studies in animals from this laboratory disclosed that systemically administered alendronate is rapidly taken up by bone tissues to the extent of 60% to 70% of the dose and excreted by the kidney, 30% to 40% in 24 hr, and that renal excretion is the only route of elimination. This study was designed to explore the effect of calcium on plasma protein binding and the renal handling of alendronate. The binding of alendronate to rat plasma was concentration, pH and calcium dependent. The fraction of unbound drug in rat plasma increased from about 3% to 9% over a drug concentration range of 0.2 to 10 micrograms/ml. Supplementation of calcium strongly augmented the binding to serum albumin. The binding of alendronate in plasma increased with increasing pH from about 50% at pH 6.6 to 98% at pH 8.6. The effects of pH on the binding of calcium and of alendronate to serum albumin were qualitatively similar. Under steady-state conditions, the binding of alendronate was substantially lower in hypocalcemic rats but unchanged in hypercalcemic rats. Although hypocalcemia caused a significant decrease in the renal secretion of alendronate, there was no effect on the renal secretion of tetraethylammonium bromide and p-aminohippuric acid. The differential effect of hypocalcemia suggests that calcium may play an important role in the renal handling of alendronate. However, hypercalcemia resulted in a substantial decrease of renal secretion of all three compounds and the decreased renal secretion was associated with a marked decrease in the glomerular filtration rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246141 TI - Renovascular actions of angiotensin II in the isolated kidney of the rat: relationship to lipoxygenases. AB - Several actions of angiotensin II have been linked to metabolism of arachidonic acid by lipoxygenases. To evaluate the importance of this interaction intrarenally we tested the effect of three different lipoxygenase inhibitors, BW755c (50 microM), a dual lipoxygenase-cyclooxygenase inhibitor, MK447 (200 microM), a nonselective lipoxygenase inhibitor which can stimulate cyclooxygenase, and baicalein (1 microM), a highly selective 12-lipoxygenase inhibitor, on angiotensin II-evoked hemodynamic changes in the rat isolated kidney, perfused with oncotic agents. Kidneys were pretreated with indomethacin (10 microM) to exclude participation of cyclooxygenase-dependent arachidonate products. Renal perfusion pressure was kept constant at 90 mm Hg by continuous adjustments in perfusate flow rate. Inhibition of cyclooxygenase alone produced a transient potentiation of the vasoconstrictor response to angiotensin II without altering GFR. On the other hand, the lipoxygenase inhibitors attenuated the angiotensin II-induced increase in renal vascular resistance by approximately 50% and promoted an increase in GFR above that of kidneys infused with angiotensin II in the presence of only indomethacin. Base-line values were essentially unchanged by lipoxygenase inhibition. Furthermore, the vasoconstrictor response to the thromboxane/endoperoxide agonist U46619 was unaffected. We conclude that products of the lipoxygenase pathway, arising within the kidney, contribute to the renal hemodynamic effects of angiotensin II. PMID- 8246142 TI - Passage of pituitary adenylate cyclase activating polypeptide1-27 and pituitary adenylate cyclase activating polypeptide1-38 across the blood-brain barrier. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) is a newly discovered regulatory peptide related to vasoactive intestinal peptide and is found widely distributed throughout peripheral tissues and the central nervous system. We examined the ability of its two major forms of 38 (P38) and 27 (P27) amino acid residues to cross the murine blood-brain barrier. After i.v. injection, [125I]P27 had a unidirectional influx constant (Ki) into the brain of 2.13 (10(-3)) ml/g/min with no saturable component to entry. Despite being larger and less lipophilic, [125I]P38 entered the brain more than 30% faster by a saturable transport system. A peptide-T analog related to PACAP that has its own saturable transport system did not inhibit the entry of [125I]P38, but did alter its binding to vascular receptors. Despite the greater Ki of [125I]P38, a larger percentage of the i.v. dose of [125I]P27 entered the brain due to favorable pharmacokinetics. However, [125I]P38 was more resistant to degradation within the brain and, after correction for degradation, its Ki increased to 16.5 (10(-3)) ml/g/min. The influences of peripheral degradation and sequestration by capillaries were negated by use of the brain perfusion and capillary depletion methods. These showed that the Ki into the brain interstitial fluid/parenchymal space for [125I]P38 was 15.3 (10(-3)) ml/g/min and was again inhibited with unlabeled P38. Both PACAPs were transported out of the central nervous system and inhibited the efflux of the other, but there was a [125I]P38 preferring subcomponent to the transport system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246143 TI - Enrichment for metallothionein does not confer resistance to cisplatin in transfected NIH/3T3 cells. AB - Evidence has been presented both that metallothionein does and does not produce resistance to cisplatin. The metallothionein-enriched cells described in most previous studies have been selected for resistance to heavy metals, such as cadmium, or have been maintained in a medium enriched for the metals. Exposure to toxic metals could alter the cells in many ways. This report addresses the effect of metallothionein content alone, independent of exposure to metals, on cellular resistance to cisplatin. The toxicity of cisplatin was compared in NIH/3T3 cells that vary in their content of metallothionein as a consequence of transfection with a plasmid that results in the constitutive expression of metallothionein. The plasmid contains the bovine papillomavirus genome and the mouse metallothionein-I gene; it is driven by a glucose-regulated protein of 78 kD. Control cells were transfected with a similar plasmid in which the coding sequences for metallothionein were inverted and separated from the promoter, thereby abolishing expression. Expression of metallothionein required neither selection nor maintenance of cells in the presence of heavy metals. Despite large differences between the two types of cells in their cellular content of metallothionein and in their resistance to the toxicity of cadmium, no differences in resistance to cisplatin were observed. PMID- 8246144 TI - Suppression of peritoneal macrophage phagocytosis of Candida albicans by opioids. AB - Receptor-selective opioid agonists have been found to suppress the capacity of macrophages to ingest opsonized sheep erythrocytes. In an effort to characterize the immunomodulatory activity of opioids further, experiments were done to examine the uptake of Candida albicans by opioid-treated murine peritoneal macrophages. It was found that treatment with morphine and selective mu, i.e., DAMGO, delta, i.e., DPDPE, and kappa, i.e., trans-3,4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl) cyclohexyl]benzene-acetamide methanesulfonate, receptor agonists resulted a concentration-dependent suppression of both the percentage of phagocytic cells and the average number of ingested yeasts. Antagonists selective for mu, i.e., H-D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2, delta, i.e., naltrindole, and kappa, i.e., norbinaltorphimine, opioid receptors completely blocked the respective receptor-selective agonist-induced suppression. These results suggest that the mu, delta and kappa opioid receptors can modulate macrophage function. PMID- 8246145 TI - Differential regulation of neuropeptide Y systems in limbic structures of the rat. AB - Neuropeptide Y-like immunoreactivity (NPYLI) in the frontal cortex and nucleus accumbens was significantly decreased after acute and multiple administrations of phencyclidine-HCl (PCP). The role of dopamine, serotonin and sigma receptors in these PCP-induced effects was evaluated. Neither the dopamine D1 antagonist SCH 23390 nor the D2 antagonist sulpiride by itself altered cortical neuropeptide systems, but in combination they totally blocked the PCP-induced changes. In contrast, sulpiride alone significantly decreased accumbens NPYLI content and enhanced the PCP-induced decreases, whereas SCH 23390 alone had no effect on accumbens NPYLI levels but did attenuate PCP-induced effects. Neither depletion of serotonin nor blockage of the sigma "receptor" had any effect on PCP-induced changes in either structure. The effects of the selective, noncompetitive N methyl-D-aspartate receptor antagonist MK-801 on cortical and accumbens NPYLI content were similar to those of PCP, suggesting an N-methyl-D-aspartate receptor mechanism in these effects. Administration of gamma-aminobutyric acid transaminase (GABA-T) inhibitors, gamma-vinyl-GABA (GVG, vigabatrin, MDL 71,754) or aminooxyacetic acid alone had no effect on cortical NPYLI content; however, administration of aminooxyacetic acid alone decreased accumbens NPYLI levels. Co administration of these GABA-T inhibitors with PCP completely blocked PCP-induced cortical NPYLI decreases and attenuated NPYLI changes in the accumbens. These data suggest that limbic neuropeptide systems are differentially modulated by N methyl-D-aspartate and dopaminergic activity and that glutamatergic influences on cortical and accumbens NPY systems are mediated, at least in part, by GABAergic mechanisms. PMID- 8246146 TI - Facilitated dissociation of albumin-fatty acid complexes by rat hepatocytes. AB - Experiments were conducted comparing palmitate clearance by hepatocyte monolayers, polyethylene and that predicted by the diffusion-reaction model. The objective was to learn whether the albumin-dependent enhancement of unbound palmitate clearance was wholly attributable to codiffusion of bound and unbound fatty acid across an unstirred fluid layer, or whether the uptake flux was also contributed by facilitated dissociation of albumin-fatty acid complexes at the cell surface. We use a model-independent formulation of the facilitation phenomenon to explore this question at low concentrations of albumin. The central finding that facilitation occurs takes into consideration albumin-dependent changes in solution viscosity, endogenous albumin secretion and the adsorption of albumin to the polyethylene surface. PMID- 8246147 TI - Chelation of zinc antagonizes the neuromuscular blocking properties of the seven serotypes of botulinum neurotoxin as well as tetanus toxin. AB - Botulinum neurotoxin types A, B (unactivated and activated), C, D, E, F and G, as well as tetanus toxin, paralyzed transmission in mouse phrenic nerve hemidiaphragm preparations. Toxin-induced blockade of transmission was antagonized by chelators [e.g., ethylenediamine tetraacetic acid, tetrakis(2 pyridylmethyl)ethylenediamine or diethylene-triaminepentaacetic anhydride], but this effect was dependent on incubation conditions. Pretreatment of toxin with chelators failed to produce antagonism, but pretreatment of tissues did produce antagonism. Of the various chelators tested, tetrakis(2 pyridylmethyl)ethylenediamine produced the greatest effect. Antagonism of toxin induced neuromuscular blockade could be partially reversed by washing chelators from tissues and could be fully reversed by adding an excess of zinc. The ability of chelators to antagonize clostridial neurotoxins was specific and did not extend to phospholipase A2 neurotoxins. Ligand-binding studies with radioiodinated toxin and brain membrane preparations showed that chelators did not antagonize toxicity by inhibiting toxin association with receptors. Similarly, pharmacological experiments with unlabeled toxin- and type-specific antibodies demonstrated that chelators did not act by blocking receptor-mediated internalization of toxin. The chelators appeared to exert their effects by antagonizing the intracellular actions of clostridial neurotoxins. Electrophysiological studies showed that chelators, at concentrations relevant to antagonism of botulinum neurotoxin and tetanus toxin, did not enhance transmitter release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246148 TI - Competitive inhibition by capsazepine of [3H]resiniferatoxin binding to central (spinal cord and dorsal root ganglia) and peripheral (urinary bladder and airways) vanilloid (capsaicin) receptors in the rat. AB - Capsazepine was reported to block capsaicin- and resiniferatoxin (RTX)-induced responses both in vivo and in vitro with Schild plots suggesting a competitive mechanism of action. We have used the [3H]RTX binding assay, thought to represent the vanilloid (capsaicin) receptor, to explore the inhibitory mechanism of capsazepine at the receptor level in the rat. In competition assays, capsazepine inhibited [3H]RTX binding by spinal cord, dorsal root ganglion (DRG) and urinary bladder membranes with similar Ki values of 4.0 +/- 0.3, 3.5 +/- 0.5 and 5.0 +/- 1.0 microM (mean +/- S.E.M.; three determinations), respectively. By contrast, capsazepine was 35- to 50-fold more potent for inhibiting specific [3H]RTX binding in the airways (Ki = 0.12 +/- 0.02 microM; mean +/- S.E.M.; four determinations). In experiments in which the concentration of [3H]RTX was varied, 10 microM capsazepine reduced the affinity of the vanilloid receptor expressed by DRG and spinal cord membranes for [3H]RTX from 15 +/- 3 to 43 +/- 5 pM, and from 20 +/- 3 to 80 +/- 5 pM (mean +/- S.E.M.; three determinations), respectively, without a measurable change in Bmax or in cooperativity index; these shifts in affinity yield Ki values of 5.2 and 3.3 microM for DRG and spinal cord membranes, respectively. Capsaicin inhibited [3H]RTX binding by spinal cord, DRG and urinary bladder membranes with a 6- to 13-fold higher potency than did capsazepine; the Ki values were 0.3 +/- 0.1, 0.6 +/- 0.4 and 0.5 +/- 0.2 microM (mean +/- S.E.M.; three determinations), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246149 TI - Divergent pharmacologic activities of PD 132301-2 and CL 277,082, urea inhibitors of acyl-CoA:cholesterol acyltransferase. AB - The in vitro potencies and hypocholesterolemic properties of CL 277,082 and PD 132301-2, two urea inhibitors of acyl-CoA:cholesterol acyltransferase (ACAT) were compared. PD 132301-2 was several-fold more potent at inhibiting ACAT in microsomes from rat and rabbit tissues and in cultured cells (murine macrophages and the human HepG2 cell line). This disubstituted urea was also relatively specific for ACAT as other cholesterol esterifying enzymes (e.g., lecithin:cholesterol acyltransferase, pancreatic cholesterol ester hydrolase), as well as intestinal diglyceride synthesis (acyl-CoA:monoglyceride acyltransferase), were unaffected in vitro at relevant concentrations. In normal chow-fed rats, both compounds reduced plasma triglycerides at doses > 50 mg/kg, but only PD 132301-2 reduced plasma cholesterol. In rat and rabbit models of hypercholesterolemia the greater in vitro potency of PD 132301-2 translated into greater in vivo efficacy (i.e., ED50 values 2- to 3-fold higher for CL 277,082 in both acute and chronic rate models). Of particular note was the greater elevation of high-density lipoprotein-cholesterol and parenteral activity of PD 132301-2 compared to CL 277,082 in the chronic rat model. Inhibition of cholesterol absorption in rats was also greater with PD 132301-2. In guinea pigs, in which 77% of plasma cholesterol was transported in low-density lipoprotein, PD 132301-2 potently reduced plasma total cholesterol (lowest significant dose = 1 mg/kg) as well as plasma triglycerides. CL 277,082 only reduced cholesterol at doses > 100 mg/kg in this low-density lipoprotein model. In a canine model of hypercholesterolemia CL 277,082 was inactive at doses up to 50 mg/kg, but PD 132301-2 was active at 3 mg/kg. Moreover, efficacy in dogs with PD 132301-2 was positively correlated with plasma drug concentration, an observation not previously demonstrated for other hypolipidemic drugs. The combined data illustrate that pharmacologic activities can vary widely among ACAT inhibitors of the same general class. In addition, the unique observation of proportionality between efficacy and blood drug levels in nonrodent animal models may not only help to simplify early stages in drug development but also may help to predict or monitor a direct action of the drug on vascular disease. PMID- 8246150 TI - Insulin is a superior antidote for cardiovascular toxicity induced by verapamil in the anesthetized canine. AB - Because of its positive inotropic effects that are independent of cyclic AMP, insulin was compared to epinephrine and glucagon as a novel treatment for cardiac toxicity from verapamil. Twenty-four alpha-chloralose-anesthetized mongrel canines of either sex were instrumented to monitor standard hemodynamic and cardiodynamic parameters and maximum elastance at end systole, via the transit time technique, as our index of contractility. Toxicity was induced by 0.1 mg/kg/min of verapamil (i.v.), until 50% reduction in mean arterial blood pressure or complete AV dissociation for 30 min. This was followed by continuous infusion of 1.0 mg/kg/hr of verapamil during one of four treatment protocols: 1) control (0.9% NaCl, 2.0 ml/min); 2) epinephrine (1.0 micrograms/kg/min); 3) hyperinsulinemic-euglycemic (HIE) clamp (recombinant insulin at 4.0 U/min with 20% dextrose, arterial glucose clamped); or 4) glucagon (0.2-0.25-mg/kg bolus infusion followed by 150-micrograms/kg/min infusion). Treatments were continued until death or 240 min after which time surviving animals received a 3.0-mg/kg additional bolus of verapamil. Verapamil decreased all hemodynamic parameters during titration. All controls died within 85 min. All treatments tended to improve hemodynamics; however, HIE significantly improved maximum elastance at end systole, left ventricular end diastolic pressure and coronary artery blood flow vs. other treatments (P < .05, repeated measures). Glucagon transiently restored sinus rhythm (four animals), but in all cases reverted to A-V dissociation, coincident with sharp decreases in circumflex artery blood flow and contractility.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246151 TI - Transport of 2,4,5-trichlorophenoxyacetic acid across the blood-cerebrospinal fluid barrier of the rabbit. AB - Transport of the anionic herbicide, 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) across the blood-cerebrospinal fluid barrier was investigated using the isolated choroid plexus of the adult rabbit in vitro and ventriculocisternal perfusion in vivo. In vitro, 2,4,5-T transport was effective, with tissue concentrations 20 times those in the medium after only 5 min of incubation with 1 microM 2,4,5-T. The tissue to medium ratios reached steady state by 20 min at approximately 45 fold. Uptake was energy dependent and inhibited by ouabain, phloridzin and several organic anions (probenecid, 2,4-dichlorophenoxyacetic acid and octanoate). Neither tyrosine (transported by a separate system) nor the neurotoxin, quinolinic acid, inhibited 2,4,5-T transport. Kinetic analysis yielded an apparent Km of 58 microM and Vmax of 111 nmol g-1 min-1 in the lateral ventricular choroid plexus with similar values (57 microM and 87 nmol g-1 min-1) in the fourth ventricular plexus. In vivo, the steady-state clearance of 2,4,5-T from the cerebrospinal fluid exceeded that of inulin and was reduced in a dose dependent fashion by 2,4-dichlorophenoxyacetic acid and probenecid. Together, these data indicate that 2,4,5-T is cleared from the brain and cerebrospinal fluid by the organic anion transport system and that alterations in such transport may have a significant impact on the toxicity of this agent in the central nervous system. PMID- 8246152 TI - Mechanism-based inactivation of cytochrome P450 1A1 by N-aralkyl-1 aminobenzotriazoles in guinea pig kidney in vivo and in vitro: minimal effects on metabolism of arachidonic acid by renal P450-dependent monooxygenases. AB - Guinea pig renal microsomes convert arachidonic acid to two classes of P450 dependent metabolites, epoxyeicosatrienoic acids (EET), and 16- through 20 hydroxyeicosatetraenoic acids [(16-20)-OH-AA)]. The rate of formation of these metabolites was not altered by beta-naphthoflavone induction, which increased P450 1A1-dependent 7-ethoxyresorufin O-deethylation activity approximately 100 fold. alpha-Naphthoflavone, which inhibits renal P450 1A1 in vitro, did not inhibit the formation of these metabolites in microsomes from induced animals. In induced animals, N-benzyl-1-aminobenzotriazole and N-alpha-methylbenzyl-1 aminobenzotriazole, administered i.v., inhibited microsomal 7-ethoxyresorufin O deethylation activity by approximately 50% without inhibiting the formation of either class of arachidonic acid metabolites. In vitro these mechanism-based inhibitors inactivated 1A1 by > 90% without inhibiting EET or (16-20)-OH-AA formation. These data show that P450 1A1 does not bioactivate arachidonic acid to either (16-20)-OH-AA or EET in guinea pig kidney, and that N-benzyl-1 aminobenzotriazole and N-alpha-methylbenzyl-1-aminobenzotriazole selectively inactivate P450 1A1 in comparison to the P450 isozyme(s) that metabolize arachidonic acid in the kidney. In guinea pig liver beta-naphthoflavone treatment, which induces P450 1A1 and 1A2, increased the rate of the formation of (16-20)-OH-AA and EET and in vitro alpha-naphthoflavone inhibited the formation of these metabolites in induced hepatic microsomes by approximately 50 and approximately 35%, respectively. These data demonstrate that a beta naphthoflavone-inducible isozyme, most likely 1A2, converts arachidonic acid to both (16-20)-OH-AA and EET in guinea pig liver. PMID- 8246153 TI - Effects of nicotine on neocortical electrical activity in rats. AB - The present study investigates the effects of acute and repeated nicotine i.p. treatment on cortical EEG activity. Nicotine at 0.3 and 0.9 mg/kg, but not at 0.1 mg/kg, decreased high voltage spindles (HVSs). Nicotine at 2.7 mg/kg suppressed HVSs completely. Mecamylamine, a nicotinic cholinergic antagonist, increased HVSs at 5 and 7.5 mg/kg. Nicotine blocked the HVS induction induced by mecamylamine. Mecamylamine at 1.25 mg/kg antagonized the HVS suppressing action of nicotine at 0.3 mg/kg. The muscarinic cholinergic antagonist, scopolamine (0.2 mg/kg), increased the 1 to 20 Hz amplitude sum value, and this increase was blocked to some extent by the highest dose of nicotine (2.7 mg/kg). However, nicotine did not block the effect of a higher scopolamine (2.0 mg/kg) dose on the sum amplitude values. Mecamylamine at 2.5 and 7.5 mg/kg blocked the effect of nicotine at 2.7 mg/kg on the EEG sum amplitude values in scopolamine (0.2 mg/kg) treated rats. The peripherally acting nicotinic and muscarinic cholinergic antagonists, hexamethonium and scopolamine methylbromide, had no effect on spectral EEG and HVS values. In quisqualic acid nucleus basalis-lesioned rats, a frontal cortical choline acetyltransferase depletion (-72%) and slowing of the EEG was observed. Nicotine could not restore EEG activity in nucleus basalis lesioned rats. After repeated (10 days, three injections/day) administration of nicotine, no tolerance to the effects of either nicotine (0.9 mg/kg) on spontaneously occurring HVSs or nicotine (2.7 mg/kg) on the EEG change induced by scopolamine was observed. The present results show that nicotinic receptor stimulation desynchronizes neocortical EEG activity in normal animals, but this action disappears in basal forebrain-lesioned animals. Therefore, it is likely that the effects of nicotine in reversing EEG and behavioral abnormalities observed in Alzheimer's disease may be limited if the basal forebrain cell loss is extensive. PMID- 8246154 TI - Influence of 2,3-dimercaptopropane-1-sulfonate (DMPS) and meso-2,3 dimercaptosuccinic acid (DMSA) on the renal disposition of mercury in normal and uninephrectomized rats exposed to inorganic mercury. AB - The effects of the water-soluble chelating agents 2,3-dimercapto-1-propane sulfonate (DMPS) and meso-2,3-dimercaptosuccinic acid (DMSA) on the renal disposition of inorganic mercury were studied in normal and uninephrectomized (NPX) rats injected (i.v.) with a nontoxic 0.5-mumol/kg dose of mercuric chloride (HgCl2). When a 100-mg/kg dose of either DMPS or DMSA was injected (i.p.) 24 and 30 hr after treatment with HgCl2, the renal concentration and burden of inorganic mercury decreased markedly in both normal and NPX rats during the 24 hr after the first dose of the respective chelating agent was administered. Treatment with DMPS was more effective than treatment with DMSA in reducing the renal burden of mercury in both groups of rats. The fall in the renal concentration and burden of mercury in both normal and NPX rats was due primarily to a decrease in the content of mercury in the renal cortex and outer stripe of the outer medulla. However, the decrease in the concentration of inorganic mercury in the outer stripe was significantly greater in NPX rats than in normal rats. Both chelating agents caused urinary excretion of mercury to increase significantly in normal and NPX rats. In association with the increased renal release of mercury in NPX rats, the urinary excretion of mercury per gram of kidney was significantly greater in NPX rats than in normal rats. These data indicate that the renal handling of DMPS and DMSA may be altered significantly after a substantial reduction in renal mass. Findings from the present study also show that treatment with DMPS, but not with DMSA, causes the content of mercury in the liver and cellular fraction of blood to decrease in normal and NPX rats. These findings indicate that there are significant differences in the extrarenal handling of these two chelating agents. The findings in the present study suggest that DMPS and DMSA are very effective agents in reducing the renal (and whole body) burden of inorganic mercury in normal and NPX rats. PMID- 8246155 TI - Prevention of experimental carotid artery thrombosis by applaggin. AB - Applaggin, an inhibitor of platelet aggregation via binding to the glycoprotein IIb/IIIa receptor, was examined in an anesthetized canine model of arterial thrombosis formation secondary to arterial wall injury. Both carotid arteries were isolated and instrumented with flow probes, intravascular anodal electrodes and adjustable constrictors. The right carotid artery was injured initially and served as the control response to vessel wall injury in each animal, whereas the left carotid was injured after applaggin administration (1.0 mg/kg, i.v.). Arterial occlusion in the control vessel occurred in each of seven animals. Time for occlusive thrombus development was 125.6 +/- 15.2 min. One of the seven left carotid arteries occluded after applaggin. Thrombus weight was greater in control vessels (44.2 +/- 7.0 mg) vs. thrombus weight after applaggin (11.2 +/- 2.4 mg). Cyclic flow variations occurred in all control arteries before development of an occlusive thrombus. In contrast, cyclic flow variations were observed only in two of seven vessels injured after applaggin. One of the latter vessels developed an occlusive thrombus. Platelet counts, heart rate and blood pressure were unaltered over the course of the experimental protocol. Ex vivo platelet aggregation to arachidonic acid was examined before and after applaggin administration. Platelet rich plasma from animals having initial normal baseline aggregation no longer aggregated 30 min after administration of applaggin. Aggregation returned to normal within 3 hr. Applaggin binds to stimulated and unstimulated platelets and two classes of binding sites were identified. The results demonstrate that applaggin possesses an antithrombotic effect in the experimental model of canine carotid artery thrombosis. PMID- 8246156 TI - Types A and B monoamine oxidase contribute to the metabolism of norepinephrine in perfused lungs of rats. AB - The aim of the study was to determine the activity of monoamine oxidase (MAO) and the contributions of the A and B forms of MAO to the metabolism of norepinephrine (NE) in isolated perfused lungs of the rat. Preliminary experiments to investigate the MAO-A and MAO-B selective inhibitors, Ro41-1049 and Ro19-6327, respectively, were carried out in isolated preparations of rat vas deferens incubated in 10 nM [3H]NE (Uptake2 and catechol-O-methyltransferase inhibited) to measure rates of deamination of NE and the rate constant for deamination (kMAO) by MAO-A. These experiments showed that 1) the IC50 of Ro41-1049 for inhibition of MAO-A was 9.62 nM with 97% inhibition by 300 nM and 2) 100 or 300 nM Ro19-6327 did not affect deamination of NE by MAO-A. Hence, 300 nM Ro41-1049 and 100 nM Ro19-6327 were used to inhibit selectively MAO-A and MAO-B, respectively, in rat lungs perfused in vitro with 10 nM [3H]NE. Catechol-O-methyltransferase was inhibited and kMAO values for metabolism of NE were determined. The kMAO value for deamination of NE in the absence of Ro41-1049 or Ro19-6327 was 0.186 min-1. Ro41-1049 and Ro19-6327 decreased this kMAO value by 79 and 33%, respectively. Thus, it was concluded that both MAO-A and MAO-B contribute to the deamination of NE in the pulmonary circulation of the rat, with MAO-A responsible for 67-79% of the total deamination and MAO-B for 21-33%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246157 TI - Caffeine withdrawal: apparent heterologous sensitization to adenosine and prostacyclin actions in human platelets. AB - Chronic exposure to caffeine increases the number of adenosine receptors (up regulation) but these observations have been mostly limited to animal models that study A1 adenosine receptors. The regulation of adenosine receptors by caffeine in humans and, in particular A2 receptors, remains largely unexplored. The purpose of this study was to test the hypothesis that withdrawal from chronic caffeine administration results in up-regulation of A2 adenosine receptors in humans. The authors also wanted to determine whether caffeine induces homologous or heterologous up-regulation. Caffeine 250 mg three times daily was given orally to a total of 19 normal volunteers for 7 days. Platelets were obtained at base line and 12 and 60 hr after the last dose of caffeine and the antiaggregation responses to adenosine and prostacyclin receptors were evaluated ex vivo. Plasma caffeine levels remained elevated at 22 microM 12 hr after the last dose but decreased to 0.6 microM at 60 hr. Adenosine receptor activation with the agonist 5'-N-ethylcarboxamidoadenosine and prostacyclin receptor activation with iloprost or prostaglandin E1 produced a greater antiaggregation effect at 60 hr postcaffeine. Increased responsiveness to both receptors could also be demonstrated at 12 hr after removal of caffeine by washing the platelets. Sensitization to the actions of prostacyclin, however, was reversed if caffeine was added ex vivo. These results support the hypothesis that chronic caffeine exposure induces heterologous up-regulation of adenosine and prostacyclin receptors in humans and implies that endogenous adenosine normally modulates platelet adenosine receptors in vivo. These findings may be relevant to the caffeine withdrawal syndrome observed in humans. PMID- 8246158 TI - Anticonvulsant doses of 2-chloro-N6-cyclopentyladenosine, an adenosine A1 receptor agonist, reduce GABAergic transmission in different areas of the mouse brain. AB - The possible relationship between A1 adenosine receptors and the gamma aminobutyric acid (GABAA) receptor complex was evaluated in the mouse brain. We studied the effect of in vitro addition and in vivo administration of 2-chloro-N6 cyclopentyladenosine (CCPA), the most selective ligand for A1 receptors, on the biochemical parameters used currently to evaluate GABAergic function. In vitro, CCPA (0.01-100 microM) failed to modify [3H] GABA binding, [3H]flunitrazepam binding, t-[35S]butylbicyclophosphorothionate ([35S]TBPS) binding and muscimol stimulated 36Cl- uptake. On the contrary, in vivo, CCPA (1.4-27.6 mumol/kg i.p.) increased [35S]TBPS binding in membranes from the cerebral cortex, hippocampus, striatum and substantia nigra, but not from the cerebellum, thalamus, hypothalamus and olfactory tubercle. The specific A1 receptor antagonist 8 cyclopentyl-1,3-dipropylxantine (9.8 mumol/kg i.p.) abolished the effect of CCPA on [35S]TBPS binding, indicating that the action of this compound is mediated by its interaction with A1 receptors. Diazepam (1.7 mumol/kg i.p.), a positive modulator of GABAergic transmission, antagonized the increase of [35S]TBPS binding induced by CCPA. CCPA (2.8-8.3 mumol/kg i.p.) antagonized convulsions induced by isoniazid, an inhibitor of GABA synthesis, but neither antagonized nor potentiated isoniazid-induced increase of [35S]TBPS binding. CCPA (2.8-8.3 mumol/kg i.p.) antagonized the convulsions induced by pentylenetetrazol (398 mumol/kg i.p.), methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (50 mumol/kg i.p.) and bicuculline methiodide (9.8 mumol/kg i.p.). The results show that, in spite of its anticonvulsant activity, CCPA reduces the function of the GABA-coupled chloride channel function. This finding suggests that the anticonvulsant target sites are different from those involved in the action of CCPA on GABAA receptors. PMID- 8246159 TI - A novel, potent and selective nonpeptidic delta opioid receptor agonist BW373U86. AB - Four different opioid receptor binding assays and three different isolated tissue studies were used to screen for delta receptor-selective nonpeptidic compounds. (+/-)-4-((alpha-R*)-alpha-((2S*,5R*)-4-Allyl-2,5- dimethyl-1-piperazinyl)-3 hydroxybenzyl)-N,N-diethylbenzamide (BW373U86) was a potent delta receptor selective ligand in receptor binding assays. The Ki values were 1.8 +/- 0.4, 15 +/- 3, 85 +/- 4 and 34 +/- 3 nM for delta, mu, epsilon and kappa receptor binding sites, respectively. BW373U86 inhibited electrically evoked muscle contraction of mouse vas deferens with an ED50 value of 0.2 +/- 0.06 nM. This inhibitory effect of BW373U86 was antagonized by the delta receptor-selective antagonist naltrindole in a competitive manner: the Schild plot indicated a slope of 1 and a pA2 value of 9.43 (Ke = 3.7 x 10(-10) M), which is consistent with the high affinity of naltrindole in delta receptors. BW373U86 did not interact significantly with other receptors. BW373U86 inhibited the acoustic startle reflex after subcutaneous administration from 0.2- to 2-mg/kg doses in rats, and this inhibition was blocked by naltrindole. BW373U86 also induced a dose dependent increase of locomotor activity in rats at similar doses. This effect was inhibited by naltrindole. These data suggest that BW373U86 is a potent and selective nonpeptidic delta agonist, and it elicits distinct in vivo pharmacological activities. PMID- 8246160 TI - Antinociceptive actions of BW373U86 in the mouse. AB - This study explored the antinociceptive properties of (+-)-4-[(alpha- R*)-alpha [(2S*,5R*)-4-allyl-2,5-dimethyl-1-piperazinol]-3-hydroxy benzyl] - N,N-diethyl benzamide dihydrochloride (BW373U86) a nonpeptidic compound proposed to be a delta opioid agonist, using three models of nociception and four routes of administration in mice. BW373U86 produced dose- and time-dependent, naloxone sensitive antinociception in both the tail-flick and tail-pinch assays when given intrathecally (i.t.). However, at doses up to 187 nmol/mouse, i.c.v. BW373U86 was inactive in the tail-flick or tail-pinch assays. Additionally, at doses up to 187 mumol/kg, BW373U86 was not active after i.p. or p.o. administration in these endpoints. Further, in the tail-flick test, i.c.v. BW373U86 did not antagonize the antinociceptive effects of i.c.v. [D-Pen2,D-Pen5]enkephalin or [D Ala2,Glu4]deltorphin, but partially antagonized the effects of i.c.v. morphine. In the acetic-acid abdominal constriction assay, BW373U86 produced dose-dependent antinociceptive effects when given by the i.p., i.c.v. or i.t., but not by the p.o., routes. Intrathecal BW373U86 was 663-fold more potent in the abdominal constriction assay than when given by the same route in the tail-flick test. The effects of an A70 dose of i.p. or i.c.v. BW373U86 in the abdominal constriction assay were partially antagonized by i.c.v. naloxone, but not by i.c.v. N,N diallyl-Tyr-Aib- Aib-Phe-Leu-OH, where Aib is alpha-aminoisobutyric acid (ICI 174,864) or naltrindole. In contrast, i.t. naloxone, ICI-174,864 or naltrindole antagonized the antinociceptive effect of i.p. or i.t. BW373U86 in the abdominal constriction assay.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246161 TI - Discriminative stimulus effects of BW373U86: a nonpeptide ligand with selectivity for delta opioid receptors. AB - Several opioid agonists were evaluated in pigeons trained to discriminate i.m. injections of sterile water from either the mu agonist morphine (5.6 mg/kg), the kappa agonist bremazocine (0.032mg/kg) or (+/-)-4-((alpha-R*)-alpha-((2S*,5R*)-4 allyl-2,5-dimethyl-1- piperazinyl)-3-hydroxybenzyl)-N,N-diethylbenzamide (BW373U86; 0.56 mg/kg). Pigeons were trained to peak one of two keys on a fixed ratio 20 schedule for food reinforcement. The pattern of substitution of mu, kappa and delta selective agonists in the three groups of birds suggested that the discriminative stimulus effects of morphine, bremazocine and BW373U86 were different; however, a component of the discriminative stimulus effects of BW373U86 appeared to be shared with morphine. Apparent pA2 values for naltrexone with morphine, bremazocine and BW373U86 were 7.6, 6.8 and 6.3, respectively. The apparent pA2 value for naltrindole with BW373U86 was 8.3. Naltrindole (10.0 mg/kg) produced a 3-fold shift to the right in the dose-effect curve for morphine but did not antagonize bremazocine. Although results from the substitutions experiments suggested that a component of the BW373U86 discriminative stimulus was mediated through mu opioid receptors, the fact that naltrindole was 1000-fold more potent and naltrexone was 30-fold less potent in antagonizing BW373U86 than morphine indicated that the discriminative effects of BW373U86 were also mediated through delta opioid receptors. PMID- 8246162 TI - A novel delta opioid agonist, BW373U86, in squirrel monkeys responding under a schedule of shock titration. AB - The squirrel monkey titration procedure was used to assess the antinociceptive effects of the novel delta opioid agonist (+/-)-4-(a-R*)-a(2S*,5R*)-4-allyl-2,5 dimethyl-1-piperazinyl)-3-hydroxyb enzyl)- N,N-diethylbenzamide (BW373U86). Under this procedure shock increased every 15 sec from 0.01 to 2.0 mA in 30 steps. Five responses terminated the shock for 15 sec, after which the shock resumed at a lower intensity. The intensity at which the monkeys maintained the shock 50% of the time (median shock level, MSL) was determined. BW373U86 (1.0-30.0 mg/kg i.m.) increased MSL, but these increases were not dose-dependent and lasted only 15 min or less. Doses of BW373U86 that increased MSL often produced tremors and/or convulsions immediately after administration. When 1.0 mg/kg of naltrindole, a delta opioid antagonist, was given in combination with BW373U86, doses of BW373U86 up to 30 mg/kg no longer increased MSL nor did tremors and/or convulsions occur. Doses of BW373U86 (0.01-0.3 mg/kg i.m.) that did not increase MSL when administered alone shifted the dose-effect curve for the mu agonist l methadone to the left. These shifts were antagonized dose-dependently by naltrindole. In monkeys that were tolerant to morphine, BW373U86 (0.03-0.1 mg/kg i.m.) shifted the morphine dose-effect curve leftward. In addition, BW373U86 altered the effects of the partial opioid agonists, buprenorphine, nalbuphine, butorphanol and levallorphan such that doses of these drugs that did not increase MSL when administered alone, often did so in the presence of BW373U86. Taken together, these findings indicate that BW373U86 has a delta agonist profile in the squirrel monkey; however, its antinociceptive effects in the shock titration procedure may be due to its toxic effects. PMID- 8246163 TI - A nonpeptidic delta opioid receptor agonist, BW373U86, attenuates the development and expression of morphine abstinence precipitated by naloxone in rat. AB - The effect of (+-)-4-((alpha-R*)-alpha-((2S*,5R*)-4-allyl-2,5-dimethyl-1- piperazinyl)-3-hydroxybenzyl)-N,N-diethylbenzamide (BW373U86), the first potent nonpeptidic, highly selective delta opioid receptor agonist, on morphine dependence was studied in rats. Continuous infusion of BW373U86 by a subcutaneously implanted osmotic minipump did not induce any abnormal behavior. After 6 days of BW373U86 infusion, intraperitoneal injection of a high dose of naloxone or naltrindole did not precipitate morphine-like abstinence syndromes. Furthermore, a single injection of BW373U86 did not induce abstinence syndromes or modulate morphine abstinence precipitated by naloxone in chronic morphine treated rats. However, naloxone-precipitated abstinence syndromes in morphine dependent rats were partially suppressed by BW373U86 in a dose-dependent manner when the compound was infused subcutaneously before and throughout morphine treatment. Abstinence signs such as wet-dog shake, forelimb tremor and teeth chattering were either suppressed or the intensity was significantly attenuated in these BW373U86-infused rats. This effect was antagonized by naltrindole. These data show that chronic infusion of BW373U86 does not produce physical dependence and that it attenuates some abstinence behaviors in morphine-dependent rats via delta opioid receptors. PMID- 8246164 TI - Convulsive effects of systemic administration of the delta opioid agonist BW373U86 in mice. AB - A systemically active, nonpeptidic delta receptor-selective agonist, (+-)-4 ((alpha-R*)-alpha-((2S*,5R*)-4-allyl-2,5-dimethyl-1-piperazinyl) -3- hydroxybenzyl)-N,N-diethylbenzamide (BW373U86), produced a brief, nonlethal convulsion in mice. The behavioral pattern of convulsion produced by pentylenetetrazol was similar to that produced by systemic administration of BW373U86. Although several episodes of convulsion occurred with pentylenetetrazol, BWB373U86 produced a single, brief episode. Naltrexone (10.0 and 100 mg/kg) and naltrindole (1.0, 3.2 and 10.0 mg/kg), but not midazolam (0.32 mg/kg), produced dose-dependent rightward shifts in the potency of BW373U86 to induce a convulsion. A dose of 3.2 mg/kg of midazolam completely eliminated convulsions induced by BW373U86. Midazolam (0.32 and 3.2 mg/kg), but not naltrindole (3.2 and 32.0 mg/kg), produced parallel rightward shifts in the pentylenetrazol dose-effect curve. Pretreatment with a single injection of BW373U86 (3.2, 10.0, 32.0 or 100 mg/kg) produced a dose-related reduction in the capacity of BW373U86 to induce a second convulsion. Recovery of sensitivity to BW373U86 did not return to control levels for up to 2 weeks after pretreatment with a single injection of 32.0 mg/kg of BW373U86. Naltrindole (3.2 mg/kg) administered within 1 hr, but not at 2 hr, after a pretreatment dose of 10.0 mg/kg of BW373U86 prevented the refractoriness (tolerance) induced by the single dose of BW373U86. These data suggest that the convulsions as well as the tolerance induced by BW373U86 were initiated through delta opioid receptors. PMID- 8246165 TI - Effects of opioid agonists selective for mu, kappa and delta opioid receptors on schedule-controlled responding in rhesus monkeys: antagonism by quadazocine. AB - Rhesus monkeys were trained to respond under a fixed-ratio 30 schedule of food reinforcement. The mu opioid agonists alfentanil and fentanyl, the kappa opioid agonists ethylketocyclazocine (EKC) and U69,593, the delta opioid agonist BW373U86 [(+-)-4-((R*)-a-((2S*5R*)-4-allyl-2,5-dimethyl-1-piperazinal)-3-h ydroxy benzyl)-N,N-diethylbenzamide dihydrochloride] and the nonopioid, noncompetitive N-methyl-D-aspartate antagonist ketamine all produced a dose-dependent decrease in rates of responding. Quadazocine (0.1-10 mg/kg) antagonized the rate decreasing effects of all the opioid agonists, but not of ketamine. The in vivo apparent pA2 values (95% CL) for quadazocine in combination with each agonist were: alfentanil, 7.7 (7.6-7.8); fentanyl, 7.7 (7.6-7.8); EKC, 6.3 (5.9-6.7); U69,593, 6.5 (5.9-7.0); and BW373U86, 5.5 (5.3-5.8). Additionally, antagonist effects of individual doses of quadazocine in combination with each agonist were evaluated by using in vivo apparent pKB analysis, and pKB values were found to be similar to the more rigorously determined pA2 values. The relative pA2 and pKB values of quadazocine in antagonizing the rate-decreasing effects of mu, kappa and delta opioid agonists corresponded to the relative potency of quadazocine in displacing the specific binding of the mu agonist [3H]Tyr-D-Ala-Gly-(Me)-Phe-Gly ol (IC50 = 0.080 nM), the kappa agonist [3H]U69,593 (IC50 = 0.52 nM) and the delta agonist [D-Pen2,D-Pen5]-[3H]enkephalin (IC50 = 4.6 nM) from binding sites in membranes from monkey brain cortex.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246166 TI - Effects of naftidrofuryl on hypoxia-induced activation and mortality of human endothelial cells. AB - The present study was designed to elucidate the possible beneficial effects of naftidrofuryl on ischemia-induced endothelium damage. For this purpose, an in vitro model was developed wherein human endothelial cells isolated from umbilical vein were submitted to hypoxia. Long-term hypoxia incubation (6 h) induced cell mortality, and naftidrofuryl strongly protected endothelial cells against this mortality in a dose-dependent manner and at concentrations as low as 10(-9) M. 66% protection was still observed after 16 h of hypoxia. Naftidrofuryl had to be present during the hypoxia incubation to exert its action; preincubation up to 24 h in the presence of naftidrofuryl could not protect endothelial cells incubated under hypoxia without naftidrofuryl. Short-term hypoxia, which does not induce mortality, strongly activates the endothelial cells with an increase in the cytosolic calcium concentration, in the phospholipase A2 activity, and in the synthesis of prostaglandin and of platelet-activating factor. It also enhances the adherence of polymorphonuclear neutrophils. Naftidrofuryl was able to markedly inhibit this whole cascade of events in a dose-dependent manner. We also demonstrated that naftidrofuryl could block the decrease in ATP concentration that results from the hypoxic conditions. These results indicate that by preserving the energetic level of the cells, naftidrofuryl prevents the activation of endothelial cells and the cell mortality induced by hypoxia. By maintaining an intact endothelium in vivo during ischemia, naftidrofuryl could prevent the further damage induced by leukocyte recruitment and activation. PMID- 8246167 TI - Impact of L-dopa on striatal acetylcholine release: effects of 6-hydroxydopamine. AB - We investigated the effects of the dopamine (DA) precursor L dihydroxyphenylalanine (L-DOPA) on electrically evoked acetylcholine (ACh) overflow from rat striatal slices. Some animals were pretreated 1 to 2 months earlier with 6-hydroxydopamine, (6-OHDA), a catecholamine neurotoxin, so as to selectively destroy DA terminals (98.6% striatal DA depletion). Although the addition of L-DOPA (10 microM) produced a 37% inhibition of ACh overflow in slices from lesioned rats, it failed to affect ACh overflow in slices from intact animals. In contrast, ACh overflow from intact slices exposed to L-DOPA and to the DA uptake inhibitor nomifensine (1 microM) was 22% greater than in the presence of nomifensine without L-DOPA. ACh overflow from slices prepared from lesioned rats was 45% greater with both drugs than in the presence of nomifensine by itself. Superfusion with the aromatic L-amino acid decarboxylase (AADC) inhibitor NSD-1055 (250 microM) abolished the inhibitory effects of L-DOPA, as did L-sulpiride (1 microM), an inhibitor of DA receptors of the D2 subtype. These results suggest that inhibition of ACh overflow by L-DOPA is mediated by DA formed from exogenous L-DOPA which then acts on D2 receptors. They further indicate that the net impact of the loss of nigrostriatal terminals is an increased dopaminergic inhibition of striatal cholinergic interneurons in response to exogenous L-DOPA. This appears to result in large part from a lesion induced reduction in high-affinity reuptake of DA formed from exogenous L-DOPA. PMID- 8246168 TI - Insulin-like growth factor-I ameliorates transient ischemia-induced acute renal failure in rats. AB - Acute renal failure in rats was induced by transient occlusion of bilateral renal arteries and veins to investigate whether insulin-like growth factor-I (IGF-I) has an effect on the damaged renal function or not. Administration of IGF-I at 0.01, 0.1 and 1 mg/kg by s.c. injection caused a 18.7, 33.0 and 66.5% increase of glomerular filtration rate and 54.8, 61.2 and 84.1% decrease of blood urea nitrogen, respectively, compared with the values in the saline-treated group 2 days after ischemia. Other renal parameters tested such as fractional excretion of sodium, N-acetyl-beta-D-glucosaminidase and tubular reabsorptance of phosphorus which are thought to represent renal function of proximal and distal tubules, respectively, were also improved by IGF-I treatment. A histochemical study also supported these observations. Severe epithelial necrosis of proximal tubules and decrease of brush borders were observed 2 days after transient ischemia in the saline-treated group, whereas marked histochemical alterations were not observed in the IGF-I-treated group. L-NG-nitroarginine, an inhibitor of nitric oxide synthetase, prevented the improvement of glomerular filtration rate and blood urea nitrogen by IGF-I at 1 mg/kg, suggesting that the ameliorative action on renal function by IGF-I is mediated via nitric oxide, possibly its vasodilating action. These findings provide the first evidence for the efficacy of IGF-I in the model of acute renal failure, suggesting that IGF-I may be useful for the treatment of acute renal failure. PMID- 8246169 TI - Enhanced expression of rat hepatic CYP2B1/2B2 and 2E1 by pyridine: differential induction kinetics and molecular basis of expression. AB - Expression of the cytochrome P450 (CYP) 2B subfamily in rat and rabbit hepatic tissues after pyridine (PY) treatment has been examined, and the molecular basis for enhanced 2B1/2B2 expression has been determined. P450 expression was monitored using metabolic activity, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analyses, and the identity of the proteins was confirmed through N-terminus microsequence analysis. PY caused a dose-dependent elevation of hepatic CYP2B1/B2B levels in rats, which ranged from 4- to 22-fold over the dosing regimen of 100 to 400 mg PY/kg/day, for 3 days, respectively. PY at low dose failed to induce CYP2B in rabbit hepatic tissue, suggesting a species dependent response in 2B expression. Anti-2B1 IgG addition to PY-induced microsomes inhibited benzphetamine N-demethylase activity by only approximately 15%, in sharp contrast to the approximately 73% inhibition observed for phenobarbital-induced microsomes, suggesting the induction of other form(s) of P450 having benzphetamine N-demethylase activity. Northern blot analysis revealed that PY treatment increased 2B1 and 2B2 poly(A)+ RNA levels approximately 69- and approximately 34-fold, respectively, whereas the 2E1 poly(A)+ RNA levels failed to increase. The results of this study show that PY induces CYP2B1/2B2 and that induction is species-dependent and kinetically distinguishable from 2E1 induction. Moreover, 2B1/2B2 induction occurs as a result of elevated mRNA levels associated with either transcriptional activation or mRNA stabilization, and it differs from the mechanism of hepatic 2E1 induction by PY. PMID- 8246170 TI - A study of the dose dependency of a glycine receptor antagonist in focal ischemia. AB - N-methyl-D-aspartate receptor antagonists are potent neuroprotectants in experimental focal cerebral ischemia, but behavioral and neuropathologic changes seen with these drugs in rodent models may limit the clinical utility of these compounds. Glycine's modulation of N-methyl-D-aspartate channel function offers another pharmacologic approach to excitotoxicity in ischemia. The potent glycine antagonist 7 Chlorothiokynurenic acid (7-Cl-Thio-Kyna) was studied in a permanent middle cerebral artery occlusion stroke model in the rat. The compound was effective, in a dose-dependent manner, in attenuating infarct size when administered before or after permanent middle cerebral artery occlusion. Its activity was mainly due to glycine antagonism inasmuch as 5 Chlorothiokynurenic acid, a compound having other pharmacological activities in common with 7-CI-Thio Kyna (for instance the radical scavenger action), was inactive in this model. 7 Cl-Thio-Kyna did not produce cytological changes similar to MK 801. PMID- 8246171 TI - [3H]BIMU-1, a 5-hydroxytryptamine3 receptor ligand in NG-108 cells, selectively labels sigma-2 binding sites in guinea pig hippocampus. AB - The binding of [3H]endo-N-(8-methyl-8-azabicyclo[3.2.1.]oct-3-yl)- 2,3-dihydro-3 ethyl-2-oxo-1H-benzimidazole-1-carboxamide hydrochloride ([3H]BIMU-1) a benzimidazolone with high affinity for 5-hydroxytryptamine (5-HT)3 and 4 5-HT3 and 5-HT4 receptors, was characterized in NG-108 cells and guinea pig hippocampus. Specific, heat-sensitive, binding of [3H]BIMU-1 was detected in both NG-108 cells and guinea pig hippocampus. In NG-108 cell membranes, a portion of the specific binding was displaced by 5-HT3 receptor ligands with affinities and specificity consistent with the labeling of 5-HT3 receptors. The residual specific binding was insensitive to serotonin (Ki > 1 mM) but was displaced by haloperidol (Ki of 50 nM). In guinea pig hippocampal membranes [3H]BIMU-1 binding was insensitive to serotonin but was displaced by haloperidol, and 1,3-di-o-tolyl guanidine with affinities appropriate for the labeling of a sigma binding site (Ki of 6.3 and 31 nM, respectively). The affinity profile of ligands displacing [3H] BIMU-1 binding in guinea pig hippocampus was consistent with the selective labeling of a sigma-2 binding site because the sigma-1 selective benzomorphans, (+)-pentazocine and (+)-N-allylnormetazocine, only weakly displaced the binding (Ki greater than 1 microM). The affinity of BIMU-1 for sigma-2 binding sites (Ki = 32 nM) was 200-fold greater than that for sigma-1 binding sites (Ki = 6.3 microM), dopamine (D1 and D2), other serotonin (5-HT1A, 5-HT2A, 5-HT2C) and muscarinic (M1, M2, M3 and M4) receptors (Ki > 10 microM). The distribution of haloperidol-sensitive [3H]BIMU-1 binding was also consistent with the labeling of sigma-2 binding sites. These data suggest that [3H]BIMU-1 selectively labels sigma-2 binding sites in guinea pig hippocampus. [3H]BIMU-1, under appropriate experimental conditions, is thus the first sigma-2 binding site radioligand to be characterized. PMID- 8246172 TI - Autoradiographic distribution of [14C]tetrahydrobiopterin and its developmental change in mice. AB - Tetrahydrobiopterin (BH4) is a regulatory factor of phenylalanine degradation as well as of catecholamine, serotonin and nitric oxide synthesis. To understand the in vivo metabolism of BH4, whole-body autoradiography was performed on mice at various developmental stages after injection of a low dose (45 micrograms/kg) of radiolabeled BH4. In adult mice, high levels of radioactivity were accumulated in the liver and kidney, suggesting that BH4 in these organs is supplied not only by intracellular de novo biosynthesis but also by uptake from the blood. In contrast, little radioactivity was found in the brain, adrenal medulla and bone marrow, in which high levels of endogenous BH4 are found, suggesting that BH4 in these organs is supplied mainly by intracellular biosynthesis. Biopterin, a fully oxidized form of BH4, was not accumulated in any tissues and was excreted rapidly. In pregnant mice, maternal BH4 passed through the placenta and was distributed uniformly to the fetal tissues. Neonatal (5- and 7-day-old) mice had a similar distribution pattern of labeled BH4 to that of adult mice, except for the liver and kidney: in the liver, selective accumulation of BH4 in the adult was not observed in 5-day-old mice; in the kidney, the distribution of high radioactivity to the inner cortex observed in the adult was found in neither 5- nor 7-day-old mice. The developmental increase in hepatic BH4 accumulation was correlated with those of the activities of phenylalanine hydroxylase and GTP cyclohydrolase I, the rate-limiting enzyme of the BH4 biosynthetic pathway. PMID- 8246173 TI - Role of the complement system in antigen-induced bronchoconstriction and changes in blood pressure in the guinea pig. AB - Systemic anaphylaxis involves life-threatening bronchoconstriction and a serious hypotensive response often complicated by cardiac arrhythmias. The purpose of the present study was to determine whether complement system activation is essential to the bronchoconstriction and the changes in blood pressure seen in guinea pig models of anaphylaxis. The soluble complement receptor 1 (sCR1; BRL 55730) was used to inhibit activation of the classical and alternative pathways of complement in the guinea pig, and to determine whether this inhibition prevents bronchoconstriction and changes in blood pressure induced by i.v. antigen injection in guinea pigs that are either passively or actively sensitized to the antigen ovalbumin. sCR1 at 15 mg/kg did not affect significantly either the antigen-induced bronchoconstriction or the changes in blood pressure in a guinea pig passively sensitized with immunoglobulin G antibody to ovalbumin. However, it shortened the duration of the antigen-induced increase in blood pressure and inhibited the antigen-induced decrease in circulating platelets in an actively sensitized guinea pig. Continued studies using a cumulative dose of sCR1 of 105 mg/kg administered over a 24-hr period demonstrated that sCR1 attenuated the bronchoconstrictor response and the decrease in circulating platelets and prevented the hypotension induced by antigen in an actively sensitized guinea pig. At a cumulative dose of 105 mg/kg, sCR1 did not inhibit the bronchoconstrictor or blood pressure response to either histamine or bradykinin, indicating that its attenuation of cardiovascular and respiratory reactivity is specific for complement-related processes. The anaphylactic response was accompanied by complement activation as evidenced by cleavage of the C3 molecule. In the presence of sCR1, no C3 cleavage products were detectable in the plasma. Our studies demonstrate that complement activation is an essential step in the antigen-induced bronchoconstriction and the changes in blood pressure in an actively sensitized guinea pig model of anaphylaxis. Continued studies of the differing mechanisms and mediators of anaphylaxis are of importance, and the complement system clearly warrants consideration as a source of those mediators. PMID- 8246174 TI - Selective expression of methotrexate-resistant dihydrofolate reductase (DHFR) activity in mice transduced with DHFR retrovirus and administered methotrexate. AB - To determine the effect of in vivo pharmacological selective pressure on the insertion and expression of new gene sequences, retroviral-mediated transfer of a methotrexate-resistant dihydrofolate reductase (Mtxr-DHFR) gene in hematopoietic tissue was investigated using a murine syngeneic bone marrow transplant system. A series of recombinant retroviral vectors were constructed to contain long terminal repeat (LTR) regions from different murine retroviruses associated with various proliferative disorders of the lymphohematopoietic system, including Moloney leukemia virus, spleen focus-forming virus (anemia strain) and myeloproliferative sarcoma virus. High-titer DHFR virus (10(7) colony-forming units/ml) was generated by gene amplification adapting virus-producer cell lines to grow in medium containing increasing concentrations of Mtx. This high-titer DHFR virus was used to introduce the Mtxr-DHFR gene into murine hematopoietic tissue by injecting DHFR virus-exposed marrow into lethally irradiated syngeneic recipient mice that subsequently were administered Mtx. Southern blot analysis of spleen DNA demonstrated insertion of the DHFR provirus in all surviving mice transplanted with DHFR virus-exposed marrow. However, enzymatic assay of crude spleen extracts demonstrated the presence of Mtxr-DHFR activity only in mice that were administered Mtx; nonadministered animals or animals transplanted with control (neo) virus-infected marrow contained undetectable drug-resistant enzyme activity. These results suggest the selective outgrowth of hematopoietic tissue harboring and expressing a DHFR provirus in animals administered Mtx and have implications for the application of drug-resistance gene insertion in somatic tissues of animals and humans. PMID- 8246175 TI - Correlation between insulin receptor occupancy and tyrosine kinase activity at low insulin concentrations and effect of major histocompatibility complex class I derived peptide. AB - In rat adipocytes, insulin dose-response curves were determined for the following effects in the same cells under the same conditions: glucose uptake, binding to insulin receptors (IR), IR autophosphorylation in vivo and in vitro, IR tyrosine kinase activity and insulin-stimulated phosphatidylinositol (PI) kinase. All the EC50 values were essentially the same (mean +/- S.E.M. was 7 +/- 1 nM), except for glucose uptake, which was 170 pM. Using an improved method, we were able to measure PI kinase activity at picomolar concentrations of insulin (> 30 pM) corresponding to the EC50 for glucose uptake. These experiments showed that insulin-stimulated increase in glucose uptake was associated with an increase in antiphosphotyrosine antibody precipitable PI kinase activity, consistent with the view that IR tyrosine kinase activity may be involved in insulin-mediated signaling of glucose uptake. Small peptides (17-25 residues long) derived from major histocompatibility complex class I have previously been shown to inhibit IR internalization without any effect on the affinity of insulin to the receptor. It is now shown that the peptide-mediated inhibition of internalization, which doubles the number of insulin-occupied receptors at an insulin concentration of 70 pM, also results in a corresponding enhancement of PI kinase activity and glucose uptake. Thus, the receptors arrested on the cell surface by the peptide are biologically active. PMID- 8246176 TI - Endothelial control of the pulmonary circulation in normal and chronically hypoxic rats. AB - 1. The effect of blockade of nitric oxide synthesis in pulmonary endothelium by two L-arginine analogues was tested in isolated blood-perfused lungs of normal rats and rats exposed chronically to 10% O2. 2. In both groups of rats the analogues (N-monomethyl-L-arginine (L-NMMA) and N-nitro-L-arginine methyl ester (L-NAME)) enhanced hypoxic vasoconstriction. In normal rats, with rare exceptions, these analogues had little or no effect on pulmonary artery pressure (Ppa) at constant blood flow during normoxia. However, chronically hypoxic rats have pulmonary hypertension and in these rats the analogues always raised Ppa; the rise in Ppa after L-NMMA but not L-NAME could be partially reversed by L arginine. L-NAME was more potent than L-NMMA. 3. To see whether the difference between rat groups was due to the high Ppa in chronically hypoxic rats, in control rats we raised Ppa passively by lung inflation to values higher than found in chronically hypoxic rats. L-NAME did not alter the effects of lung inflation on Ppa. 4. Ppa was also raised passively by plotting pressure-flow lines up to high flow rates; the lines were changed minimally by both analogues in control rats but in chronically hypoxic rats the lines were raised to higher pressures and steepened substantially. 5. In control rats, during vasoconstriction caused by hypoxia, endothelin 1 and almitrine, L-NAME caused further rises in pressure. We conclude that a stimulus for nitric oxide release in control rats is the narrowing of vessels caused by vasoconstriction rather than passive increases in intravascular pressure. 6. In chronically hypoxic rats arterioles are narrowed by growth of new muscle and there is some muscle tone even in normoxia. Thus narrowing of the vascular lumen is the stimulus common to both groups of rats which leads to nitric oxide synthesis and attenuation of Ppa by a negative feedback process. Narrowing is associated with a large increase in shear stress due to two factors; the pressure drop along a vessel segment is increased and the surface area of the lining of the affected segment is decreased. 7. Atrial natriuretic peptide caused dose-dependent pulmonary vasodilation in both rat groups but had a greater effect in chronically hypoxic rats. The action persisted and was enhanced after blockade of NO synthesis. PMID- 8246177 TI - Influence of stimulation parameters on the release of adenosine, lactate and CO2 from contracting dog gracilis muscle. AB - 1. The addition of adenosine, CO2 and lactate to the venous blood draining an isolated constant-flow perfused gracilis muscle was studied in anaesthetized and artificially ventilated dogs during twitch and tetanic contractions. 2. Venous adenosine concentration increased from 154 +/- 33 nM (mean +/- S.E.M.) to 279 +/- 121 or 280 +/- 125 nM after 10 min of 1.5 or 3 Hz twitch contractions and to 240 +/- 120 or 276 +/- 139 nM after 10 min of 1 or 5 s tetani occurring at 0.1 Hz. Twitch contractions at 0.1 or 0.5 Hz for 10 min did not significantly elevate venous adenosine. 3. Venous lactate concentration was significantly increased after 10 min of 1.5 or 3 Hz twitches or 5 s tetani at 0.1 Hz. There was a good correlation (r = 0.70; P < 0.001) between venous adenosine and lactate concentrations. 4. Venous partial pressure of CO2 (PCO2) was significantly elevated after 10 min of 1.5 or 3 Hz twitch contractions or 1 or 5 s tetani at 0.1 Hz. There was also a good correlation (r = 0.58; P < 0.001) between venous adenosine concentration and PCO2. 5. Venous partial pressure of O2 (PO2) decreased during all contractions except those at 0.1 Hz, but the oxygen cost per unit of tension x time was similar during every pattern of stimulation, and the percentage of the total energy production achieved by anaerobic means during muscle contractions did not exceed that at rest, indicating that there had been no limitation to the oxygen supply. Venous PO2 was poorly correlated with venous adenosine concentration (r = 0.28), but quite well correlated with venous lactate concentration (r = 0.53; P < 0.001). If the indirect influence of PO2 on venous adenosine concentration via an increase in lactate concentration was eliminated by partial correlation, then the coefficient for the relationship between venous adenosine concentration and venous PO2 became 0.15. 6. There was a significant correlation between the venous adenosine concentration and the venous pH (r = 0.53; P < 0.001). If the influence of oxygenation on venous adenosine and pH was eliminated by partial correlation, the coefficient for the relationship between venous adenosine and pH increased to 0.95.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246178 TI - Force relaxation, labile heat and parvalbumin content of skeletal muscle fibres of Xenopus laevis. AB - 1. Measurements were made of stable (hb) and labile (ha) maintenance heat rate, slowing of relaxation as a function of tetanus duration, and parvalbumin (PA) content in intact single muscle fibres of types 1 and 2 from Xenopus laevis. The majority of experiments were performed at 20 degrees C. In addition, total and myofibrillar ATPase activity was measured in skinned Xenopus fibres, also of types 1 and 2; these studies were performed at 4 degrees C. 2. In agreement with a previous study hb was significantly higher in type 1 (175 +/- 13 mW (g wet wt) 1; n = 8) than in type 2 fibres (88 +/- 9 mW (g wet wt)-1; n = 7). The value of ha was 236 +/- 22 and 117 +/- 16 mW (g wet wt)-1, respectively (mean +/- S.E.M.). ha decayed with a time constant of 0.27 +/- 0.02 (n = 8) and 0.33 +/- 0.02 s (n = 7). 3. The early relaxation rate of tetanic force, extrapolated to the onset of stimulation (yo + yb; where yo is 'extra' rate of relaxation and yb steady rate) was 85.6 +/- 4.2 s-1 for type 1 fibres (n = 8) and 62.7 +/- 7.3 s-1 for type 2 fibres (n = 7). Relaxation rate at the end of a 1.8 s tetanus (yb) was 29.4 +/- 1.6 and 33.3 +/- 1.5 s-1, respectively; thus, there was more slowing with tetanus duration in type 1 fibres. The time constant for slowing of relaxation with tetanus duration was similar to that for decay of ha. 4. Parvalbumin concentration, [PA], was 0.45 +/- 0.04 mM in type 1 (n = 7) and 0.22 +/- 0.04 mM (n = 7) in type 2 fibres. 5. For individual fibres positive correlations were found between the 'extra' rate of relaxation (yo), labile heat (ha) and [PA]. Significantly more labile heat was liberated than can be accounted for by the enthalpy change of Ca2+ binding to PA. 6. For five fibres (type 1) studied both at 20 and 10 degrees C, the magnitude of slowing of relaxation, expressed as yo/(yo + yb), was 0.58 +/- 0.03 at 20 degrees C and 0.65 +/- 0.03 at 10 degrees C. 7. Both slowing of relaxation and labile heat were depressed in the second of two closely spaced tetani in type 1 fibres. Repriming of both effects followed similar, biphasic time courses and required more than 10 min for completion at 20 degrees C.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246179 TI - Permeability of the fetal villous microvasculature in the isolated perfused term human placenta. AB - 1. Capillary permeability-surface area (PS) products for the low molecular weight radioactive tracers, 22Na, 51Cr-EDTA (relative molecular mass 357) and 57Co cyanocobalamin (relative molecular mass 1353) were measured in the fetal circulation of isolated dually perfused lobules of normal term human placentae using the single circulation, multiple-tracer dilution technique. 2. In lobules perfused with M199 medium, containing dextran and 5 g l-1 bovine albumin, the extractions of all three tracers decreased as the flow was increased over the range of 2-8 ml min-1, and PS products for 51Cr-EDTA and 57Co-cyanocobalamin, but not for 22Na, reached constant values at flows above 0.1 ml min-1 g-1. 3. Flow independent PS products in the presence of albumin were 0.025 +/- 0.002 ml min-1 g-1 (mean +/- S.E.M., n = 25) for 57Co-cyanocobalamin and 0.057 +/- 0.003 ml min 1 g-1 (n = 25) for 51Cr-EDTA. The ratio of PS values (51Cr-EDTA/57Co cyanocobalamin) was 2.28, while the ratio of the corresponding free diffusion coefficients was 1.79, indicating substantial restriction to the diffusion of the 57Co-cyanocobalamin. 4. In another series of lobules perfused in the absence of albumin, extraction values for all three test tracers were constant over the same flow range. Values at high flow rates were therefore about twice those measured in the presence of albumin, and PS products for all three tracers failed to reach diffusion-limited values. 6. Lobules perfused with and without albumin were fixed using a glutaraldehyde fixative containing 1% Alcian Blue dye. An ultrastructural examination of the endothelium showed no significant changes in cell or cleft morphology, or in the glycocalyx, in the absence of albumin which might account for the observed permeability change. 7. These data are the first physiological measurements specifically characterizing fetal microvascular permeability in the human placenta. The results suggest that permeability resembles that found in skeletal muscle and, as such, the endothelium presents a significant barrier to the diffusion of large solutes. The observed 'protein effect' indicates that albumin can interact with elements of the solute pathway to increase its restrictiveness. PMID- 8246180 TI - Neither changes in phosphorus metabolite levels nor myosin isoforms can explain the weakness in aged mouse muscle. AB - 1. The contractile force, phosphorus metabolite levels, intracellular pH and myosin isoforms were compared in isolated soleus and extensor digitorum longus (EDL) muscles from young (6 month old) and aged (28 month old) mice, at 23 degrees C. 2. The isometric force per unit cross-sectional area was significantly lower by 21 +/- 5% in soleus muscles from aged mice compared to those from young mice (mean +/- S.E.M., n = 11 and 9 respectively). 3. The EDL muscle contained twice as much total creatine and phosphocreatine as the soleus, 1.7 times as much ATP, and 0.4 times the inorganic phosphate (Pi) per unit weight. The intracellular pH and free ADP levels were not significantly different between these muscle types. 4. There was no significant difference in resting metabolite levels between young and old EDL or soleus despite the difference in mechanical strength. 5. Examination of the expression of myosin isoforms by non-denaturing gel electrophoresis has shown that the percentage of each isoform does not change with respect to age; thus, if there is an atrophic process occurring, it is not fibre type specific. 6. We have determined that neither the Pi levels nor the intracellular pH can explain the differences seen in muscle strength with age. There is also no correlation between muscle weakness and any of the other metabolites responsible for energy transduction (phosphocreatine, ATP or ADP). PMID- 8246181 TI - Noradrenergic modulation of retinogeniculate transmission in the cat. AB - 1. Relay neurones were extracellularly recorded from the A-layers of the dorsal lateral geniculate nucleus (dLGN) of the anaesthetized cat. The noradrenergic influence on retinogeniculate transmission was investigated through microiontopheretic techniques in a total of 140 dLGN relay cells using three experimental approaches: (i) the effects of agonists for alpha 1-, alpha 2- and beta-adrenoceptors were separately analysed; (ii) the noradrenergic influence was related to the global state of activity of the relay neurones, which was associated with discrete patterns of the electroencephalogram (EEG); (iii) distinct phases of visual responses evoked from the area of the retinal receptive field, and of binocular and lateral inhibitory responses, were evaluated before, during and after the action of noradrenergic agonists. 2. The spontaneous generation of high-frequency bursts of spikes in dLGN relay neurones, associated with periods of highly synchronized, delta-like patterns of the EEG, was selectively suppressed by the beta-adrenoceptor agonist isoprenaline or the alpha 1- adrenoceptor agonist phenylephrine. Single action potentials, occurring at a low frequency between bursts, were significantly less affected. Depending upon the ejection level of the adrenoceptor agonists, burst activity was suppressed by 23-73%, compared with a reduction in single spike firing in the range 7-24%. The suppression of burst firing occurred in all functional types of dLGN relay neurones (X, Y; on, off), enhanced burst activity was observed in less than 1% of the cells. 3. On-going tonic sequences of action potentials (around 15 Hz), occurring during periods of EEG activity characterized by lower amplitudes and higher frequencies, were separately affected by adrenoceptor agonists. Isoprenaline had no significant effect, phenylephrine induced a global reduction of spike firing with no obvious relation to the ejection level, and the alpha 2 adrenoceptor agonist clonidine inhibited action potential generation in a near dose-dependent manner. 4. Visual response properties were investigated during periods of less synchronized states of EEG activity. Responses to visual stimulation of the retinal receptive field centre were not significantly influenced by isoprenaline, while phenylephrine or clonidine attenuated the phasic and the tonic response component in all functional types of relay neurones and independent of the stimulus contrast being used. At low ejection levels, slight facilitatory effects were observed with isoprenaline (65% of neurones that were tested) or phenylephrine (15%). The inhibitory influence of the antagonistic surround area of the receptive field appeared unaltered during action of isoprenaline or phenylephrine.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246182 TI - Neural induction suppresses early expression of the inward-rectifier K+ channel in the ascidian blastomere. AB - 1. Early expression of ion channels following neural induction was examined in isolated, cleavage-arrested blastomeres from the ascidian embryo using a two electrode voltage clamp. Currents were recorded from the isolated, cleavage arrested blastomere, a4-2, after treatment with serine protease, subtilisin, which induces neural differentiation as consistently as cell contact. 2. The inward-rectifier K+ current increased at the late gastrula stage shortly after the sensitive period for neural induction both in the induced (protease-treated) and uninduced cells. Ca2+ channels, characteristic of epidermal-type differentiation, and delayed-rectifier K+ channels and differentiated-type Na+ channels, characteristic of neural-type differentiation appeared much later than the inward-rectifier K+ channels, at a time corresponding to the tail bud stage of the intact embryo. 3. When cells were treated with subtilisin during the critical period for neural induction, the increase in the inward-rectifier K+ current from the late gastrula stage to the neurula stage was about three times smaller (3.67 +/- 1.74 nA, mean +/- S.D., n = 14) than in untreated cells (11.25 +/- 3.10 nA, n = 26). The same changes in the inward-rectifier K+ channel were also observed in a4 2 blastomeres which were induced by cell contact with an A4-1 blastomere. However, when cells were treated with subtilisin after the critical period for neural induction, the amplitude of the inward-rectifier K+ current was the same as in untreated cells. Thus the expressed level of the inward-rectifier K+ channel was linked to the determination of neural or epidermal cell types. 4. There was no significant difference in the input capacitance of induced and uninduced cells, indicating that the difference in the amplitude of the inward rectifier K+ currents derived from a difference in the channel density rather than a difference in cell surface area. 5. The expression of the inward-rectifier K+ channel at the late gastrula stage was sensitive to alpha-amanitin, a highly specific transcription inhibitor. In both induced and uninduced cells, injection of alpha-amanitin at the 32-cell stage reduced the current density of the inward rectifier K+ channel to about 2 nA/nF, corresponding to 13% of that recorded from uninjected cells. By contrast, the expression of the fast-inactivating-type Na+ current, which transiently increased along with the inward-rectifier K+ channel, was resistant to alpha-amanitin injection. 6. The dose of alpha-amanitin injected was controlled by monitoring co-injected fluorescent dye, fura-2.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246183 TI - Neural differentiation in cleavage-arrested ascidian blastomeres induced by a proteolytic enzyme. AB - 1. As previously reported, ectodermal a4-2 blastomeres isolated from 8-cell embryos of the ascidian, Halocynthia roretzi or aurantium, and cultured under conditions of cleavage arrest always differentiated into an epidermal phenotype, showing long-lasting Ca(2+)-dependent action potentials and/or tunic on the cell surface. a4-2 blastomeres contacted by a chordamesodermal blastomere, A4-1, differentiated into a neural phenotype, characterized by fast Na(+)-dependent spikes. Differentiation to a similar neural phenotype occurred when isolated a4-2 blastomeres from H. aurantium embryos were treated with > 0.003% subtilisin for 60 min at the 32-cell stage of the control embryo. Comparisons between induction by cell contact and induction by proteolytic enzymes were made and showed them to be similar in several respects. 2. When the serine protease, subtilisin, was used as the neural inducer, neural competence of a4-2 blastomeres, measured as the percentage frequency of the induction of Na+ spikes, increased after the 32-cell stage and decreased during the gastrula stage. The time course of the neural competence was the same as that for contact with the A4-1 blastomere. 3. The neural competence of four different ectodermal blastomeres isolated from the 16 cell embryo was also examined using subtilisin as a neural inducer, and by contact with the A4-1 blastomere from the 8-cell embryo. The competence was higher in anterior blastomeres than in posterior blastomeres for both types of induction. This regional difference in neural competence along the antero posterior axis paralleled that expected from neural cell lineage during normal development, i.e. blastomeres with more cells of neural lineage among their derivatives showed higher competence. 4. Streptomyces subtilisin inhibitor, SSI (0.1%), a specific protease inhibitor for subtilisin-type serine proteases, significantly suppressed (50%) neural induction of the ectodermal blastomere, a4 2, by contact with the chordamesodermal blastomere, A4-1. 5. Monensin, brefeldin A and bafilomycin A1, all of which affect secretory processes, suppressed the neural inducing ability of the chordamesodermal blastomere, A4-1. 6. These results permit the hypothesis that a protease secreted from the chordamesoderm generating blastomere induces the ectodermal blastomere to differentiate into neural cell type. PMID- 8246184 TI - Alterations in intracellular calcium and tension of activated ferret papillary muscle in response to step length changes. AB - 1. To study the effects of mechanical constraints on the calcium (Ca2+) affinity of cardiac troponin C, we analysed the tension and aequorin light (AL, intracellular Ca2+) transients in response to a step length change in aequorin injected ferret right ventricular papillary muscles. The muscle preparations were continuously activated with ouabain (10(-4) M) (ouabain contracture) or with high frequency stimuli in the presence of ryanodine (5 microM) (tetanic contraction). 2. The tension transient in response to either the release or stretch was oscillatory: tension decreased rapidly during the release and then increased, after which it lapsed into a new steady level in a series of damped oscillations. The opposite was true for the stretch. The oscillatory responses were conspicuous and less damped in ouabain-activated preparations (oscillation frequency of 2.2 2.3 Hz at 22 degrees and 4.5-4.6 Hz at 30 degrees C) and much more damped in ryanodine-treated preparations. 3. The transient AL response was also oscillatory, the time course of which corresponded to that of the transient tension response. Regardless of the difference in the time course of the transients in two different preparations and at two different temperatures, the increase in AL corresponded to the decrease in tension, likewise the decrease in AL to the increase in tension. 4. The mean level of AL after release was lower than the control level present just prior to the release in ouabain-activated preparations, but the AL after release finally returned to the nearly control level in ryanodine-treated preparations. 5. When the ryanodine-treated muscle was further treated with 2,3-butanedione monoxime (BDM) (20 mM), the tetanic tension decreased remarkably without affecting the AL signal. The tension transient of this preparation was quite similar to that of the resting muscle, which changed in a nearly stepwise fashion; AL was hardly affected by step length changes, as in the resting muscle, in spite of the higher AL level. 6. These results suggest that the Ca2+ affinity of cardiac troponin C is increased with an increase in tension (i.e. the cross-bridge attachment) and decreased with a decrease in tension i.e. the cross-bridge detachment), and that the mean [Ca2+]i is lowered by release, at least in a Ca(2+)-overloaded condition, mainly through the sarcoplasmic reticulum. PMID- 8246185 TI - Matching between motoneurone and muscle unit properties in rat medial gastrocnemius. AB - 1. Electrical and contractile (isometric) properties were studied for sixty-six motoneurone-muscle unit combinations from rat medial gastrocnemius (MG). The animals were anaesthetized with pentobarbitone. 2. The muscle units were classified into S (slow) and F (fast) on the basis of measurements of speed and fatigue resistance: the 'slow' category comprised units with an initial twitch contraction time exceeding those found among fatigue-sensitive units (border value 20 ms). 3. Twitch speed was assessed by three different measures: (i) contraction time (time to peak, range 11.4-28.0 ms), (ii) half-relaxation time (8.4-56.5 ms), and (iii) total twitch duration (34-116 ms). All three measures were mutually highly correlated and their respective values showed a continuous and unimodal distribution across the unit population. 4. The motoneurones were investigated with regard to their time course and amplitude of post-spike after hyperpolarization (AHP; range of total durations 30-116 ms, amplitudes 0.9-8.0 mV), rheobase (0.8-17.1 nA), input resistance (0.8-5.1 M omega) and axonal conduction velocity (33-85 m/s). 5. Motoneurones of slow-twitch muscle units (type S) had, on average, a significantly slower time course of AHP, a smaller rheobase, a higher input resistance and more slowly conducting axons than those innervating fast-twitch muscle units. 6. Across the whole neuronal sample, input conductance (reciprocal of input resistance) correlated well with rheobase (r = 0.74). However, the differences in rheobase did not seem to be caused exclusively by the associated differences in input conductance. 7. Throughout the sampled population, the relative slowness of AHP showed a continuous and highly significant correlation with the relative slowness of the corresponding unit twitch. The absolute duration of AHP was close to that of the twitch. In the Discussion it is argued that this 'speed match' between motoneurone and muscle unit would help ensure that barely recruited motoneurones start firing at a frequency that is optimally suited for the subsequent rate gradation of force. 8. AHP amplitude was, on average, significantly smaller for fast-twitch than for slow-twitch motoneurones. Calculations indicated that these differences were almost completely caused by the associated differences in input resistance; the computed value for the conductance change underlying the AHP was nearly the same for fast- and slow-twitch motoneurones. 9. A simple neurone model was used to calculate the consequences of the differences in AHP amplitude and duration for repetitive discharge properties of fast and slow cell categories.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246186 TI - Temporal and spatial summation in the human rod visual system. AB - 1. Absolute and increment thresholds were measured in a retinal region 12 deg temporal from the fovea with 520 nm targets of varying size and duration. Measurements were made under rod-isolation conditions in two normal observers and in a typical, complete achromat observer who has no cone-mediated vision. The purpose of these experiments was to determine how the temporal and spatial summation of rod-mediated vision changes with light adaptation. 2. The absolute threshold and the rise in increment threshold with background intensity depend upon target size and duration, but the psychophysically estimated dark light of the eye (the hypothetical light assumed to be equivalent to photoreceptor noise) does not. 3. The rise in increment threshold for tiny (10 min of arc), brief (10 ms) targets approaches the de Vries-Rose square-root law, varying according to the quantal fluctuations of the background light. The slope of the rod increment threshold versus background intensity (TVI) curves in logarithmic co-ordinates is about 0.56 +/- 0.04 (when cones are not influencing rod field adaptation). For large (6 deg) and long (200 ms) targets, a maximum slope of about 0.77 +/- 0.03 is attained. 4. The steeper slopes of the rod-detected TVI curves for large, long targets implies some reduction in temporal or spatial summation. In fact, the change in summation area is much more critical: under conditions where only the rod system is active the TVI curve slope is independent of target duration, suggesting that temporal summation is practically independent of background intensity. 5. The rise in threshold also depends on the wavelength of the background field in the normal observer but not in the achromat, confirming reports that the field adaptation of the rods is not independent of the quantal absorptions in the cones. The cone influence is most conspicuous on long wavelength backgrounds and is found for all target sizes and durations, but is greater for large and long targets than for the other conditions. PMID- 8246187 TI - Stimulation of the KATP channel by ADP and diazoxide requires nucleotide hydrolysis in mouse pancreatic beta-cells. AB - 1. The mechanisms by which ADP and the hyperglycaemic compound diazoxide stimulate the activity of the ATP-regulated K+ channel (KATP channel) were studied using inside-out patches isolated from mouse pancreatic beta-cells maintained in tissue culture. 2. The ability of diazoxide and ADP to increase KATP channel activity declined with time following patch excision and no stimulation was observed after 15-40 min. 3. Activation of KATP channels by ADP required the presence of intracellular Mg2+. The stimulatory effect of ADP was mimicked by AMP but only in the presence of ATP. Replacement of ATP with the non hydrolysable analogue beta, gamma-methylene ATP did not interfere with the ability of ADP to stimulate KATP channel activity. By contrast, enhancement of KATP channel activity was critically dependent on hydrolysable ADP and no stimulation was observed after substitution of alpha,beta-methylene ADP for standard ADP. 4. The ability of diazoxide to enhance KATP channel activity was dependent on the presence of both internal Mg2+ and ATP. Diazoxide stimulation of KATP channel activity was not observed after substitution of beta,gamma-methylene ATP for ATP. However, in the presence of ADP, at a concentration which in itself had no stimulatory action (10 microM), diazoxide was stimulatory also in the presence of the stable ATP analogue. 5. The stimulatory action of diazoxide on KATP channel activity in the presence of ATP was markedly enhanced by intracellular ADP. This potentiating effect of ADP was not reproduced by the stable analogue alpha,beta-methylene ADP and was conditional on the presence of intracellular Mg2+. A similar enhancement of channel activity was also observed with AMP (0.1 mM). In the absence of ATP, diazoxide was still capable of stimulating channel activity provided ADP was present. This effect was not reproduced by AMP. 6. In both nucleotide-free solution and in the presence of 0.1 mM ATP, the distribution of the KATP channel open times were described by a single exponential with a time constant of approximately 20 ms. Addition of ADP or diazoxide resulted in the appearance of a second component with a time constant of > 100 ms which comprised 40-70% of the total number of events. Under the latter experimental conditions, the open probability of the channel increased more than fivefold relative to that observed in the presence of ATP alone.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246188 TI - Repriming of L-type calcium currents revealed during early whole-cell patch-clamp recordings in rat ventricular cells. AB - 1. The establishment of the whole-cell patch-clamp recording configuration (WCR) revealed a type of inhibition to which L-type Ca2+ channels were subject in static rat ventricular myocytes before obtaining the WCR. 2. Immediately after membrane disruption (< 10 s), the Ca2+ current (ICa) was absent but gradually increased in amplitude to reach its final waveform (amplitude and kinetics) 2-3 min after the WCR was reached. 3. Three distinct phases (P) were identified. First, no inward but an outward current, blocked (1-2 min) by Cs+ dialysing from the patch pipette (P1), was recorded. Second, overlapping with (P1), ICa increased dramatically to reach a maximum peak amplitude within 2-3 min (P2). Concomitantly, its rate of decay, initially monoexponential and slow, became biexponential owing to the appearance of a fast component of inactivation (P3). Complete interconversion between slow and fast components sometimes occurred. 4. Changes in current waveform were not related to voltage loss or series resistance variation, and suppression of an outward current (P1) was unlikely to account for P2 and P3. 5. The run-up of ICa was independent of the nature of the permeating ions, the membrane holding potential, depolarization, rate of stimulation, the intracellular Ca2+, ATP, Mg2+, Cs+ and the pH of the pipette solution. Since large Ca2+ currents were recorded using the perforated patch technique, the run up of ICa is not explained by the wash-out of an inhibitory endogenous macromolecule during cell-pipette exchanges. 6. Pharmacological manipulations, including the use of Ca(2+)-Ba(2+)-EGTA and exposure of the cells to isoprenaline and/or Bay K 8644 prior to recording, did not alter the mechanism primarily responsible for build-up. Unrepriming of channel activity was required before these modulations could be effective. 7. Currents could however be instantly augmented when cells were extracellularly superfused during the run-up step. The wash-out of an inhibitory agent originating in the cell itself (such as H+, NH4+ and lactate) and accumulating in the extracellular microenvironment of the cells seems unlikely. Rather, we suggest that pressure-induced mechanostimulation may be involved in the restoration of Ca2+ channel activity. PMID- 8246189 TI - Characterization of TTX-sensitive and TTX-resistant sodium currents in small cells from adult rat dorsal root ganglia. AB - 1. The whole-cell patch-clamp technique was used to investigate the characteristics of two types of sodium current (INa) recorded at room temperature from small diameter (13-25 microns) dorsal root ganglion (DRG) cells, isolated from adult rats and maintained overnight in culture. 2. Sodium currents were isolated pharmacologically. Internal Cs+ and external tetraethylammonium (TEA) ions were used to suppress potassium currents. A combination of internal EGTA, internal F-, a low (10 microM) concentration of external Ca2+ and a relatively high (5 mM) concentration of internal and external Mg2+ was used to block calcium channels. The remaining voltage-dependent currents reversed direction at the calculated sodium equilibrium potential. Both the reversal potential and magnitude of the currents exhibited the expected dependence on the external sodium concentration. 3. INa subtypes were characterized initially in terms of their sensitivity to tetrodotoxin (TTX). TTX-sensitive (TTXs) currents were at least 97% suppressed by 0.1 microM TTX. TTX-resistant (TTXr) INa were recorded in the presence of 0.3 microM TTX and appeared to be reduced in amplitude by less than 50% in 75 microM TTX (n = 1). 4. As in earlier studies, the peak of the current-voltage relationship, the mid-point of the normalized conductance curve and the potential (Vh) at which the steady-state inactivation parameter (h infinity) was 0.5 were found to be significantly more depolarized for the TTXr INa (by ca 10, 14 and 37 mV respectively). There was little difference in the slope at the mid-point of the normalized conductance curves (the mean slope factors were 5.1 mV for the TTXs INa and 4.9 mV for the TTXr current) but the h infinity curves for TTXr currents were significantly steeper than those for TTXs currents (mean slope factors of 3.8 and 11.5 mV respectively). Both the time to peak and the decay time constant of the peak current recorded from a holding potential of -67 mV were more than a factor of three slower for the TTXr INa than for the TTXs current. 5. However, in direct contrast to the difference in activation and decay kinetics, 'slow' TTXr INa recovered from inactivation at 67mV, or reprimed, more than a factor of ten faster than 'fast' TTXs INa. 6. The differences apparent in both the repriming kinetics of TTXs and TTXr INa at -67 mV and the kinetics of the decay phase of the peak INa are shown to be explicable largely in terms of the voltage dependence of their respective inactivation systems.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246190 TI - Characterization of synaptically elicited GABAB responses using patch-clamp recordings in rat hippocampal slices. AB - 1. Tight-seal, whole-cell voltage clamp recording techniques were used to characterize monosynaptically evoked GABAB currents in adult rat brain slices maintained at 34-35 degrees C. Responses were recorded from granule cells of the dentate gyrus following the blockade of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX)-, D-2-amino-5-phosphonovaleric acid (D-AP5)- and picrotoxin-sensitive fast synaptic transmission, so that the remaining synaptic currents could be studied in isolation. 2. Under these conditions, stimulation in the molecular layer elicited a slow outward current which was blocked by the selective GABAB antagonist CGP 35348 in a concentration-dependent manner (200-800 microM). This current was absent in recordings made with pipettes containing 10-15 mM of the lidocaine derivative QX-314 or when caesium was substituted for K+. 3. Increasing the [K+]o e-fold (from 2.5 to 6.8 mM) shifted the reversal potential of the GABAB current from -97.9 to -73.2 mV, as predicted by the Nernst equation. Peak conductance was constant, but in 6.8 mM [K+]o at voltages hyperpolarized to EK (equilibrium potential for potassium), a small outward rectification was evident. 4. The time course of the current could be described by fourth-power exponential activation kinetics with double exponential inactivation. At 34-35 degrees C, the average activation time constant (tau m) was 45.2 ms, while the two inactivation time constants (tau h1 and tau h2) were 110.2 and 516.2 ms, with corresponding weighting factors (wh1 and wh2) of 0.84 and 0.16, respectively. The Q10 (temperature coefficient) values for these time constants were between 1.82 and 2.31. Neither tau m, nor tau h1 and tau h2 were voltage dependent in the range from -45 to -95 mV. 5. Paired-pulse depression of the GABAB current was studied by giving identical conditioning and test stimuli over a wide range (50-5000 ms) of interstimulus intervals (ISIs). The maximal depression (48%) occurred at 200 ms ISI, and the depression lasted for over 5 s. The magnitude of paired-pulse depression was not dependent on the postsynaptic membrane potential. 6. Application of the competitive antagonist CGP 35348 such that the peak current was diminished by approximately 50% had no effect on the activation or inactivation kinetics of the current. Similarly, during paired-pulse depression the kinetics of test currents were identical to those of conditioning currents. These findings support the hypothesis that the mechanism responsible for paired pulse depression involves a reduction in neurotransmitter release without postsynaptic alterations in K+ channel activation/inactivation kinetics.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246191 TI - Respiratory responses to electrical and chemical stimulation of the area postrema in the rabbit. AB - 1. The respiratory role of the area postrema (AP) has been investigated in pentobarbitone- or alpha-chloralose-anaesthetized, vagotomized, paralysed and artificially ventilated rabbits, by means of electrical stimulation and microinjections of DL-homocysteic acid (DLH). Phrenic nerve activity was used as an index of central respiratory drive. 2. Bipolar electrical or chemical stimulation (microinjections of DLH, 5-30 nl; 160 mM) of the caudal compact portion of the AP provoked excitatory effects on the inspiratory motor output, without apparent changes in the arterial blood pressure. 3. Depressant effects on inspiratory activity, accompanied on some occasions by changes in arterial blood pressure (as a rule, increases > or = 30 mmHg) were induced by DLH microinjections in close neighbouring areas (including the medial part of the nucleus tractus solitarii) or in the IV ventricle. 4. These results support a role for the AP in the neural control of respiration. The findings are discussed in connection with other autonomic functions to which the AP has been reported to contribute, in different animal species. PMID- 8246192 TI - Myelinated primary afferents of the sacral spinal cord responding to slow filling and distension of the cat urinary bladder. AB - 1. A total of sixty-five sacral afferent neurones with myelinated fibres supplying the urinary bladder was recorded from the sacral roots S2 in anaesthetized cats. All afferent units were identified with electrical stimulation of the pelvic nerve. The discharge properties were quantitatively evaluated using slow filling at rates of 1-2 ml min-1 and isotonic distension to preset pressure levels. Eight afferents were studied prior to and after acute sacral de-efferentation of the urinary bladder. 2. All afferent units were silent when the bladder was empty and responded in a graded manner to an increase of intravesical pressure. During slow filling the level of afferent activity correlated closely with the level of the intravesical pressure. All afferents behaved like slowly adapting mechanoreceptors with both a dynamic and static component of their discharge. With the exception of two units the intraluminal pressure threshold was below 25 mmHg. Thus virtually all myelinated afferents respond in the pressure range that is reached during a non-painful micturition cycle. 3. The stimulus-response functions of the afferents were similar regardless of whether intravesical pressure was increased by slow filling or by distension. However, during slow filling stimulation response functions often exhibited steeper slopes between 5 and 25 mmHg indicating that relatively small changes of intravesical pressure result in large changes of afferent activity. Nevertheless, all units displayed monotonically increasing stimulus response functions throughout the innocuous and noxious pressure level. 4. The stimulus response functions of the afferent neurones did not change after acute de efferentation of the urinary bladder, although the rapid phasic fluctuations of afferent activity that are produced by small contractions of the urinary bladder under normal conditions largely disappeared. This means that contractions and distension activate the afferent endings by a common mechanism. 5. It is concluded that the myelinated sacral afferents of the urinary bladder form a homogeneous population which encodes all information necessary for the normal regulation of this organ. Furthermore, this set of afferents mediates all sensations which may reach consciousness within a normal micturition cycle. PMID- 8246193 TI - Pyramidal and corticospinal synaptic effects over reticulospinal neurones in the cat. AB - 1. The spontaneous activity of 103 precruciate neurones (fifty-eight activated antidromically from the pyramidal tract but not from the corticospinal tract, PTNs; forty-five activated from both sites, CSNs) was used to trigger the average of the intracellularly recorded synaptic noise in 294 reticulospinal neurones (RSNs). These RSNs were recorded in the nucleus reticularis gigantocellularis of the contralateral medial bulbar reticular formation (NRGc) in chloralose anaesthetized cats. 2. Twelve pyramidal tract neurones (six CSNs) were tested with a single RSN, twenty-six (10 CSNs) with two RSNs each, thirty (13 CSNs) with three RSNs each, and thirty-five (16 CSNs) with four RSNs each. Postsynaptic potentials were observed in the averages generated by twenty PTNs and fifteen CSNs. 3. The only synaptic effect produced by both PTNs and CSNs upon RSNs in our sample was excitatory, and in none of the tested cases (n = 15) were any changes found in the amplitude, shape, or duration of the excitatory postsynaptic potentials (EPSPs) after injection of depolarizing or hyperpolarizing currents. This suggests that the synapses are probably located at the distal dendrites. 4. Recording of the presynaptic spike allowed separation of the conduction time and synaptic delay from the total latency. According to our data there appear to be two different types of excitation of corticofugal neurones over RSNs: a monosynaptic effect produced by both PTNs and CSNs, and a disynaptic effect produced by PTNs but not by CSNs. The disynaptic EPSPs had statistically significant slower rise times and longer widths than the monosynaptic EPSPs. PMID- 8246194 TI - Capsaicin did not evoke pain from human hand vein segments but did so after injections into the paravascular tissue. AB - 1. To see if pain from veins is mediated by C fibre endings, the C fibre stimulant capsaicin was applied intravenously, and, for comparison, paravenously and intracutaneously. 2. Capsaicin, dissolved in the fat emulsion Intralipid, was applied intravenously by continuous perfusion of vascularly isolated hand vein segments as well as by injections into occluded finger veins. Using the latter approach chemicals reach the paravascular space. 3. Pain intensities were recorded continuously with an electronically controlled visual analogue scale for deriving capsaicin concentration-pain intensity relations and the time course of pain (latencies, pain durations). 4. Capsaicin always evoked pain upon injection into skin and paravenous tissue (0.3-6.5 microM) and into occluded finger veins (3.3-33 microM), whereas it had no effect whatsoever when perfused through hand vein segments even at a concentration of 650 microM. 5. Pain intensity increased with concentration and usually reached the tolerance maximum at the fivefold threshold concentration, so that the concentration-pain intensity relations were congruent for the various routes of drug application. 6. The latencies and pain durations were independent of the capsaicin concentration, but were substantially longer with injections into occluded finger veins (latency 10-30 s, pain duration 60-120 s) than with intradermal or paravenous injections (2-9 s, 10-28 s). 7. These observations show for the first time a functional similarity between the nociceptive C fibre system of the skin and the paravascular tissues, and by inference, they dismiss the possibility that C fibre endings mediate pain in cutaneous veins. PMID- 8246195 TI - Retinal origins of the temperature effect on absolute visual sensitivity in frogs. AB - 1. The absolute sensitivity of vision was studied as a function of temperature in two species of frog (Rana temporaria, 9-21 degrees C, and Rana pipiens, 13-28 degrees C). 2. Log behavioural threshold (measured as the lowest light intensity by which frogs trying to escape from a dark box were able to direct their jumping) rose near-linearly with warming with a regression coefficient of 1.26 +/ 0.03 log units per 10 degrees C (Q10 = 18). Threshold retinal illumination corresponded to 0.011 photoisomerizations per rod per second (Rh* s-1) at 16.5 degrees C. 3. The effect of dim backgrounds on jumping thresholds suggested 'dark lights' of 0.011 Rh* s-1 at 16.5 degrees C and 0.080 Rh* s-1 at 23.5 degrees C, corresponding to Q10 = 17. 4. Response thresholds of retinal ganglion cells were extracellularly recorded in the isolated eyecup of R. temporaria. The thresholds of the most sensitive cells when stimulated with large-field steps of light were similar to the behavioural threshold and changed with temperature in a similar manner. 5. The decrease in ganglion cell 'step' sensitivity with warming consisted of a decrease in summation time (by a factor of 2-3 between 10 and 20 degrees C) and an increase in the threshold number of photoisomerizations (a decrease in 'flash' sensitivity, by a factor of 2-5 over the same interval). No effect of temperature changes on spatial summation was found. 6. Frequency-of response functions of ganglion cells indicated an 11-fold increase in noise equivalent dark light between 10 and 20 degrees C (mean values in four cells 0.009 vs. 0.10 Rh* s-1). 7. The temperature dependence of ganglion cell flash sensitivity could be strongly decreased with dim background illumination. 8. It is concluded that the desensitization of dark-adapted vision with rising temperature is a retinal effect composed of shortened summation time and lowered flash sensitivity (increased numbers of photons required for a threshold response) in ganglion cells. The desensitization bears no simple relation to the apparent retinal noise increase. PMID- 8246196 TI - Kinetic analysis of the denaturation process by alcohols of sodium channels in squid giant axon. AB - 1. The effects of several aliphatic alcohols on sodium currents were examined in the intracellularly perfused squid giant axon when the same concentration of alcohol was applied on both sides of the membrane. 2. An irreversible suppression of sodium currents, accompanied by anaesthesia at high alcohol concentration, was examined in detail using four aliphatic alcohols, that is, ethanol, 1-propanol, 1 butanol and 1-pentanol. 3. This irreversible effect seemed to be attributable to the sequential denaturation of sodium channels, because the kinetics, the current voltage relation and the sodium channel activation-voltage curve did not change after the sodium current decreased. 4. The time course of the remaining sodium conductance was measured as a function of the sum of the alcohol application time by repeating the process of applying and completely washing out alcohol. The remaining sodium conductance decayed as a function of time in a single exponential manner. This decay time constant depended strongly on the concentration of alcohol and could be assumed to be the denaturation time constant of the sodium channel. 5. The denaturation time constant decreased as the alcohol concentration increased. This time constant is proportional to the Nth power of the alcohol concentration. The N values are 4.3, 4.5, 5.8 and 7.6 for ethanol, 1-propanol, 1-butanol and 1-pentanol, respectively. This implies that alcohol molecules bind to a restricted number of specific sites in the sodium channel protein to cause the denaturation. 6. The concentration of alcohol which caused the same amount of denaturation is related to the exponential function of the carbon number of the alcohol. Considering the partition coefficient of alcohol between lipid and aqueous solution, the concentration of alcohol in the membrane which denatured half of the sodium channels in 2 h can be calculated to be 0.5 M for all alcohols. PMID- 8246197 TI - Effects of distension of the urinary bladder on the cardiovascular reflexes from the carotid baroreceptors in the dog. AB - 1. The hindlimb vasoconstrictor effects of distension of the urinary bladder were studied at different levels of input from the carotid sinus baroreceptors in the dog anaesthetized with a mixture of chloralose and urethane. 2. The vascularly isolated hindlimb was perfused at constant blood flow through its femoral artery, so that a change in pressure gradient (mean femoral arterial perfusion pressure minus mean inferior vena caval pressure) indicated a similar directional change in vascular resistance. The vascularly isolated carotid sinus regions were perfused with blood at a constant pulsatile flow. 3. Raising the carotid sinus mean perfusion pressure in randomly selected steps of 30 mmHg from 60 to 210 mmHg had little effect on heart rate unless the blood pressure was controlled, when a progressive bradycardia occurred, but caused a progressive reduction in arterial blood pressure and vasodilatation in the perfused hindlimb. Distension of the bladder at each level of carotid sinus pressure resulted in tachycardia, hypertension and hindlimb vasoconstriction. 4. The cardiac responses to bladder distension were the same at all carotid sinus pressures. When the blood pressure was controlled, however, the response was reduced at high and low sinus pressures. 5. The relationship between the carotid sinus perfusion pressure and hindlimb perfusion pressure (i.e. vascular resistance) was affected by distension of the bladder in two ways. In the one, hindlimb perfusion pressure increased by approximately the same amount at all levels of carotid sinus pressure indicating resetting of the carotid sinus baroreceptor reflex control of hindlimb vascular resistance towards vasoconstriction without change in gain of the reflex. In the other, the pressure increases were diminished at the higher levels of carotid sinus pressure indicating both resetting and an increase in gain of the reflex. 6. Both types of response occurred in the spontaneously breathing animal, in animals artificially ventilated, while pacing the heart, with the arterial blood pressure maintained constant at about 100 mmHg, and after division of the cervical vagosympathetic nerves. The frequency of occurrence of each type of response, however, varied under the different conditions. 7. The possible reasons for the two types of vascular response are discussed. PMID- 8246198 TI - Inhibitory effects of histamine and bradykinin on calcium current in smooth muscle cells isolated from guinea-pig ileum. AB - 1. Single smooth muscle cells were isolated from the longitudinal muscle layer of the guinea-pig ileum and within 10 h Ca(2+)-currents (ICa) were recorded using the whole-cell patch clamp technique. 2. Histamine (10 microMs) and bradykinin (BK, 1 microM) suppressed ICa; the effect had two phases: a rapid and transient suppression of ICa followed by a sustained suppression. Acetylcholine and substance P appeared to have similar effects but these were not investigated in detail. 3. The effects of histamine and BK on ICa were established by high intracellular concentrations of the Ca2+ buffer EGTA (30 mM) or 1,2-bis(O aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) (5 mM) in the absence of Ca2+ added to the pipette solution. When [Ca2+]i was strongly buffered to 125 or 190 nM by BAPTA-Ca2+ mixtures in the pipette the transient suppression of ICa was blocked but the sustained effect still occurred. This indicated that the transient effect was caused by a rise in [Ca2+]i. The sustained effect, in contrast, did not seem to be caused by a rise in [Ca2+]i but did show Ca2+ dependence because it did not occur if [Ca2+]i was abnormally low. 4. Application of caffeine (10 mM) to deplete stored Ca2+ or intracellular heparin (1 mM) to block the action of D-myo-inositol 1,4,5-trisphosphate (IP3) to release stored Ca2+ prevented the transient but not the sustained suppression of ICa. Heparin also blocked the transient Ca(2+)-activated K+ current in response to histamine or BK. Both transient and sustained suppressions of Ca2+ channel activity were observed in the absence of extracellular Ca2+ when current was carried mostly by Na+ ions. 5. Intracellular guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S; 10 or 100 microM) induced a gradual decline of ICa upon which transient decreases of current were superimposed. Histamine caused a larger than normal inhibition of ICa and no recovery occurred on wash-out. Intracellular guanosine 5'-O-(2 thiodiphosphate) (GDP-beta-S; 1 mM) abolished the effects of histamine and BK on ICa.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246199 TI - Feed artery role in blood flow control to rat hindlimb skeletal muscles. AB - 1. Vasomotor tone and reactivity were investigated in feed arteries of the extensor digitorum longus and soleus muscles. Feed arteries are located external to the muscle and give rise to the microcirculation within each muscle. Resting diameter was smaller in feed arteries of the soleus muscle. 2. Feed arteries of both muscles dilated to similar peak values with sodium nitroprusside. 3. Micropressure measurements demonstrated resistance to blood flow in the feed arteries supplying both muscles. Feed arteries supplying soleus muscle demonstrated greater resistance to blood flow compared to feed arteries of extensor digitorum longus muscle. 4. Greater resting tone and larger pressure drop for feed arteries of soleus muscle suggest greater range of flow control compared to feed arteries of extensor digitorum longus muscle. 5. In both muscles, feed artery diameter increased with muscle contraction (functional dilatation) and in response to transient ischaemia (reactive dilatation). The magnitude of these responses varied between muscles. 6. Feed arteries are active sites of blood flow control in extensor digitorum longus and soleus muscles of the rat. These muscles differ in fibre type and recruitment properties. Differences in feed artery reactivity may contribute to differences in blood flow between these muscles observed at rest and during exercise. PMID- 8246200 TI - Studies of brain structures involved in diffuse noxious inhibitory controls in the rat: the rostral ventromedial medulla. AB - 1. Previous electrophysiological, pharmacological and anatomical evidence had suggested a possible participation of rostral ventromedial medulla (RVM) in the supraspinal part of the loop underlying diffuse noxious inhibitory controls (DNICs). In order to test this hypothesis, two experimental series were performed during which DNICs were compared in control sham-operated rats and rats with lesions of the RVM and of an adjacent candidate for such a role, the nucleus gigantocellularis reticularis (Gi). 2. In the first experimental series, lesions were induced, in anaesthetized animals, by injections of quinolinic acid (0.3-0.8 microliter of a 360 nmol/microliter solution) into the RVM or Gi. In the control animals (n = 10), the vehicle alone (artificial cerebrospinal fluid) was injected. Histological lesion reconstructions were performed after each electrophysiological experiment. Three groups of animals were considered: in the first group (n = 5), the lesion was centered on the RVM, including the two caudal thirds of nucleus raphe magnus (NRM) and adjacent reticular areas; in the second group (n = 5), the lesions extended more rostrally and involved the rostral pole of NRM; in the third group (n = 5), the lesion extended more laterally and dorsally and included nucleus reticularis gigantocellularis pars alpha (GiA), nucleus reticularis paragigantocellularis lateralis (LPGi) and the Gi. In each case, all the neuronal cell bodies within the lesioned area were destroyed. 3. In the second experimental series, electrolytic lesions of the total rostrocaudal extent of the NRM (n = 5) were induced, in anesthetized animals, by passing cathodal current (5 mA, 8 s). In the control animals (n = 5), the electrode was lowered but current was not applied. 4. One week after lesioning, the animals were anaesthetized, paralysed, artificially ventilated and recordings were made from convergent neurones in trigeminal nucleus caudalis. These neurones gave responses due to activation of A and C fibres when percutaneous electrical stimuli were applied to their receptive fields. DNICs were triggered by immersion of each paw in a 50 degrees C water-bath. Both the general properties of the convergent neurones and the inhibitions of the C fibre-evoked responses produced by these heterotopic noxious stimuli were compared in the different groups of animals. 5. The sizes of receptive field, spontaneous activities, thresholds for C fibre-evoked responses and responses to C fibre activation were not different in the control and lesioned animals. The percentage inhibitions of the C fibre evoked responses both during and in the 44s following the conditioning periods were also very similar in the different groups of animals.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246201 TI - Calcium transients in single mammalian skeletal muscle fibres. AB - 1. We studied the transient changes in myoplasmic Ca2+ concentration under current- and voltage-clamp (double Vaseline-gap technique) in cut fibres of rat extensor digitorum longus muscle using mag-fura-2 (furaptra) as Ca2+ indicator, at 3.6-3.8 microns sarcomere length and 17 degrees C. Mag-fura-5 and fura-2 were also used in order to characterize some aspects of the Ca2+ transients. 2. The peak [Ca2+] in response to a single action potential was 4.6 +/- 0.4 microM (n = 5). The time to peak of the Ca2+ transient was 4.6 +/- 0.42 ms, with half-width of 8.2 +/- 1.5 ms, time constant of the rising phase 1.15 +/- 0.25 ms, time constant of the decaying phase 3.26 +/- 0.65 ms, and delay between action potential and Ca2+ transient 2.0 +/- 0.2 ms. 3. Ca2+ transients were studied under voltage-clamp conditions at different voltages and pulse durations. The rising phase showed a complex temporal course with a fast initial increase and a second component. Both components were separated by a plateau or a brief decrease of the Ca2+ concentration. The peak Ca2+ transient was 10.5 +/- 1.3 microM (n = 22). 4. After interrupting the pulse, Ca2+ concentration decayed exponentially. The time constant of decay of the Ca2+ transient increased with the pulse voltage and duration, reaching a maximum value at potentials more positive than +10 mV and pulses longer than 200 ms. An analysis of the decaying phases of the Ca2+ transients suggests that only the removal process operates after fibre repolarization. 5. The rate of Ca2+ release from the sarcoplasmic reticulum was calculated using the Melzer, Rios & Schneider model. The value of 17.2 +/- 3.1 micronM ms-1 (n = 10) estimated in these calculations was intermediate between those obtained by other authors from cut frog muscles (10 microM ms-1) and intact frog fibres (100 microM ms-1) using antipyrylazo III (AP III) as the Ca2+ indicator. PMID- 8246202 TI - Calcium transients and calcium release in rat fast-twitch skeletal muscle fibres. AB - 1. Calcium transients were recorded from cut segments of fast-twitch rat skeletal muscle fibres stretched to 3.7-4.0 microns per sarcomere and voltage clamped at a holding potential of -80 mV using the double Vaseline-gap technique. Calcium transients were monitored simultaneously with the two calcium indicators antipyrylazo III (AP III) and fura-2. AP III was used to record the calcium changes in response to 10-200 ms depolarizing pulses to different membrane potentials while fura-2 monitored the slow decay of the transient (during 16-20 s) and the resting calcium concentration. Experiments were performed at 14-17 degrees C. 2. For 50-100 ms depolarizing pulses calcium transients were first detected between -30 and -20 mV in a total of twenty-one fibres. The transients recorded with AP III showed a plateau for small pulses (-20 mV) and a steady increase during stronger pulses (-10 mV and more positive). Upon repolarization the transients decayed towards the baseline. The signal recorded simultaneously with fura-2 showed a continuous increase of the transient during the pulses at all membrane potentials. The amplitude of the calcium transients for the large pulses could not be followed with fura-2 due to saturation of the dye. 3. The signals obtained with both dyes were used to determine the kinetics of the calcium-fura-2 reaction inside the fibres. The mean values of the kinetic parameters were: the on rate constant (kon) = 5.1 x 10(8) M-1s-1, the off rate constant (koff) = 26 s-1, and koff/kon (KD) = 69.7 nM. 4. The fast phase of decay of the calcium transients after the pulses was studied from the records obtained with AP III. For depolarizing pulses of the same duration, the rate of decay of the transients after the pulse was slower the stronger the depolarization. For pulses to the same membrane potential, the rate of decay was slower the longer the pulse duration. Both stimulating patterns indicated saturation of the removal system in the muscle fibres due to occupancy of slowly equilibrating myoplasmic calcium binding sites by released calcium. 5. The fast phase of decay of the signals obtained with AP III was well fitted with a model of the system for removing calcium from the myofilament space. 6. The rate of calcium release (Rrel) from the sarcoplasmic reticulum was calculated once the removal system was characterized in the same fibre.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246203 TI - Control of inositol polyphosphate-mediated calcium mobilization by arachidonic acid in pancreatic acinar cells of rats. AB - 1. The patch-clamp technique of whole-cell current recording was applied to single, enzymatically isolated, rat pancreatic acinar cells to investigate the current responses evoked by internal perfusion of inositol polyphosphates (InsPx). The InsPx were included in the solution filling the recording pipette and inositol 1,4,5-trisphosphate (Ins(1,4,5)P3; 10 microM) evoked transient current responses generally of less than 1 min duration, inositol 2,4,5 trisphosphate (Ins(2,4,5)P3; 10 microM) evoked smaller current transients while inositol 1,3,4,5-tetrakisphosphate (InsP4; 10 microM) evoked no detectable current response. However, in the presence (in external bathing solution) of the phospholipase A2 inhibitor 4-bromophenacyl bromide (4-BPB; 8 microM) all three of the InsPx now evoked prolonged current responses lasting for several minutes. The current responses to all three InsPx were abolished by inclusion of the Ca2+ chelator EGTA (5 mM) in the internal, pipette-filling solution indicating that the responses are calcium dependent and reflect the effect of the InsPx in increasing intracellular Ca2+. Inositol 1,3,4,5,6-pentophosphate (InsP5) induced no current response when tested up to 20 microM in the presence or absence of 4 BPB. 2. The potentiating effect of 4-BPB on the InsPx-induced current responses was not mimicked by application of arachidonic acid (AA) oxidation inhibitors; indomethacin (20 microM), nordihydroguaiaretic acid (20 microM) or proadifen (SKF525A, 100 microM). The effects of 4-BPB were countered however, by the inclusion of 2 microM AA in the external solution. The results suggest that the 4 BPB potentiates the response by inhibiting the activity of phospholipase A2, thereby reducing the formation of AA. 3. In the presence of 4-BPB (8 microM) the InsPx-evoked responses were dose dependent with an increase in both the amplitude and speed of onset with increasing concentrations. In the presence of 4-BPB InsP4 was as efficient as Ins(1,4,5)P3 both in terms of speed of onset and amplitude of responses; the efficacy and dissociation constant (Kd) for both of these InsPx were the same at 1 microM and 45 nM respectively. Ins(2,4,5)P3 was always less effective, with an efficacy and Kd of 10 microM and 750 nM respectively. 4. If 4 BPB was applied after the current responses evoked by the InsPx were over, or if guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) was included in the recording pipette then the phospholipase inhibitor gave rise to an additional, prolonged, current response.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246204 TI - ATP-regulated K+ channels are modulated by intracellular H+ in guinea-pig ventricular cells. AB - 1. The ATP-regulated potassium channel (K+ATP) was investigated with respect to modulation by intracellular pH (pHi) by using the inside-out membrane patch clamp technique in ventricular cells isolated from the heart of the guinea-pig. Channels which had been closed by internal ATP (0.3-3 mM) were dose-dependently activated by decreasing the pHi over the range of pH 7.6-6.0. However, the channel was conversely inhibited when the pHi was further decreased below 6.0. Inwardly rectifying K+ channels were also decreased in activity when pHi fell from 7.2 to 6.0. 2. The channel activation was also observed with constant concentration of free Ca2+ (1 nM) and Mg2+ (1 mM) in the bathing solution, suggesting that a change in divalent cation concentration is not involved in channel modulation by pHi. 3. When the dose-response relations of the channel activity for ATP concentrations at different pHi were examined, the channel activity obtained at 1 microM ATP was increased by decreasing pH from 7.2 to 6.4. The half-maximal inhibition for ATP concentration at pH 7.2 and 6.4 was 20 and 40 microM, respectively, and the Hill coefficient was 2.5 in both curves. 4. In the absence of ATP, internal H+ was able to reactivate run-down channels but it had less effect on the channel as long as the activity was maintained at a higher level. The increase in the channel activity by H+ was facilitated with a proceeding of the run-down. However, after the channel was completely inactivated by a long exposure of the membrane patch to ATP-free solution, a reduction of pH could not activate the channel. 5. The decrease of pH from 7.2 to 6.4 reduced single channel conductance from 89.0 to 77.7 pS in the absence of Mg2+, whereas it reduced the conductance only at the negative membrane potentials in the presence of 2 mM Mg2+. 6. Mean open and closed times within the burst-like openings of the channel remained unaffected during the change in pHi. 7. We conclude that the cardiac K+ATP channel is modulated by a change in the intracellular pH. The channel modulation consisted of the increase in the channel activity and a decrease in the permeability. The former effect was due to the decrease in the sensitivity of the channel to ATP and the reactivation of the channel which is during the process of run-down in activity. PMID- 8246205 TI - Changes in motor unit synchronization following central nervous lesions in man. AB - 1. Single motor unit spike trains have been recorded during voluntary isometric contraction of the affected intrinsic hand muscles of patients with unilateral central nervous lesions. These have been compared with similar recordings made from the patients' unaffected hand muscles and with recordings made from the hand muscles of healthy subjects. 2. Cross-correlation analysis was performed between the times of occurrence of the motor unit spike trains. The time course of central cross-correlogram peaks constructed for normal subjects and stroke patients was used to infer properties of the underlying common EPSPs and the impulse-generating properties of the motoneurones. The results of this analysis were compared between the two groups. In addition, the size and time course of cross-correlogram peaks obtained from the patients were related both to the patients' clinical state and to their hand and fine finger function. 3. Central nervous lesions were found to result in either a narrowing or broadening of the time course of motor unit synchronization. These changes were attributed either to an increase in the size of common EPSPs with respect to synaptic noise, or to the effects of presynaptic synchronization of motoneurone inputs. 4. Longitudinal studies of motor unit discharges in the year following the stroke demonstrated, in some patients, differences in the level of motor unit synchronization. These paralleled improvements in the patients' fine motor control. Pooled data from patients with varying deficits of fine motor control confirmed that loss or reduction of motor unit synchronization was associated with a corresponding slowing in the performance of rapidly alternating finger movements. 5. The results of the present study suggest that the branched common presynaptic inputs that generate motor unit synchronization are either of corticospinal tract origin or are intimately dependent on its function. Differences in the strength and time course of motor unit synchronization are demonstrated that may reflect the altered behaviour of presynaptic inputs to motoneurones following central nervous damage in man. PMID- 8246206 TI - Effects of syntactic structure in the memory of concrete and abstract Chinese sentences. AB - Smith (1981) found that concrete English sentences were better recognized than abstract sentences and that this concreteness effect was potent only when the concrete sentence was also affirmative but the effect switched to an opposite end when the concrete sentence was negative. These results were partially replicated in Experiment 1 by using materials from a very different language (i.e., Chinese): concrete-affirmative sentences were better remembered than concrete negative and abstract sentences, but no reliable difference was found between the latter two types. In Experiment 2, the task was modified by using a visual presentation instead of an oral one as in Experiment 1. Both concrete-affirmative and concrete-negative sentences were better memorized then abstract ones in Experiment 2. The findings in the two experiments are explained by a combination of the dual-coding model and Marschark's (1985) item-specific and relational processing. The differential effects of experience with different language systems on processing verbal materials in memory are also discussed. PMID- 8246207 TI - The semantics of grammatical gender: a cross-cultural study. AB - Although most present-day scholars claim that grammatical gender has no meaning correlates, anecdotal evidence dating back to the Greeks suggests that grammatical gender carries connotative meanings of femininity and masculinity. In the present study native German speakers (tested in Germany) and native Spanish speakers (tested in Mexico) judged 54 high-frequency translation equivalents on semantic differential scales chosen to reflect dimensions of evaluation, potency, and activity. Half the words were of feminine gender in German but of masculine gender in Spanish (Type I words), and half were of masculine gender in German and of feminine gender in Spanish (Type II words). As predicted, German speakers judged Type II words higher in potency than Type I words, whereas Spanish speakers judged Type I words higher in potency than Type II words. The conclusion was that grammatical gender does affect meaning. PMID- 8246208 TI - Acute pancreatitis in cats with hepatic lipidosis. AB - The purpose of this study was to characterize the incidence, clinical features, and prognosis of acute pancreatitis in cats with hepatic lipidosis. Of 13 cats histologically diagnosed with hepatic lipidosis between July 1988, and November 1989, 5(38%) were also histologically diagnosed with acute pancreatitis. In cats with hepatic lipidosis alone, the signalment, history, physical examination, and clinicopathologic findings were generally indistinguishable from those of cats with concurrent acute pancreatitis except that cats with acute pancreatitis were more likely to be cachectic and to have coagulation abnormalities. Hepatomegaly was seen on abdominal radiographs in both groups. Of the 5 cats with concurrent acute pancreatitis, abdominal ultrasonography detected 1 cat with a hypoechoic pancreas and 5 with peritoneal effusion; those abnormalities were not seen in cats without concurrent acute pancreatitis. Cats with concurrent acute pancreatitis had only a 20% recovery rate, compared with a 50% recovery rate in cats with hepatic lipidosis alone. We conclude that cats with hepatic lipidosis should be rigorously evaluated for concurrent acute pancreatitis because of 1) the rate of disease coincidence, 2) the inability of signalment, history, physical examination, and clinicopathologic findings to adequately distinguish between hepatic lipidosis and acute pancreatitis, 3) the worse prognosis associated with concurrent acute pancreatitis, and 4) the opposing nutritional strategies for hepatic lipidosis and acute pancreatitis. PMID- 8246209 TI - Congenital esophageal hiatal hernia in the Chinese shar-pei dog. AB - Esophageal hiatal hernia was diagnosed in 11 young Chinese Shar-Pei dogs between October 1985 and July 1991. The dogs ranged in age from 2 to 11 months and included 3 females and 8 males. The most common clinical signs were regurgitation, vomiting, and hypersalivation. Physical examination was normal in 6 dogs; abnormal physical examination findings in the other 5 dogs included fever, dehydration, hypersalivation, and pulmonary wheezes and crackles. Laboratory evaluation was significant only for neutrophilia in 5 dogs. A diagnosis of hiatal hernia was made on the basis of survey thoracic radiographic and/or barium esophagram findings of displacement of the esophagogastric junction and stomach into the thoracic cavity; the diagnosis was confirmed by surgery in 9 dogs and at necropsy in 2 dogs. Megaesophagus (n = 7), gastroesophageal reflux (n = 4), and esophageal hypomotility (n = 1) were additional findings in some dogs. Aspiration pneumonia was diagnosed in 7 of the dogs. Medical therapies formulated for the therapy of presumed reflux esophagitis generally failed to resolve the clinical signs associated with the hiatal hernia. Hiatal herniae were surgically repaired in 9 of the Shar-Peis by various combinations of diaphragmatic crural apposition, fixation of the esophagus to the diaphragmatic crus (esophagopexy), and left fundic tube gastropexy. Eight of the animals survived surgery, six of which have been asymptomatic since surgery (19 to 36 months). The megaesophagus, esophageal hypomotility, and bronchopneumonia resolved in all of these dogs. PMID- 8246210 TI - Radiation therapy of canine brain masses. AB - This article evaluates the responses of 14 dogs with brain masses using orthovoltage irradiation for definitive treatment. Dogs were anesthetized for computed tomography (CT) examination, formation of head immobilization and positioning devices, radiation treatment simulation, and treatments. Total doses of 39 Gy (9 dogs) or 45 Gy (5 dogs) to the tumor were administered over 25 to 41 days. Two or three portals (parallel opposed lateral with or without a dorsal field) were used. Treatment volumes included the tumor and peritumoral edema, as determined by CT scan, and a 1-cm margin. Histopathologic diagnoses were available in 9 of 14 dogs. There were 4 meningiomas, 1 lymphosarcoma, 1 pituitary adenoma, 1 metastatic anaplastic carcinoma, 1 anaplastic oligodendroglioma and 1 dog with granulomatous meningoencephalitis. At the end of radiation therapy, 10 dogs could be evaluated for progression of clinical signs: 3 dogs deteriorated or failed to improve, and 7 dogs improved. At the time of analysis, all dogs were dead. Mean and median survival times, measured from the beginning of radiation, were 345 and 489 days, respectively. This was compared with mean survival times of 30 to 81 days reported in the literature for dogs with brain tumors that did not receive treatment. The median survival time of 9 dogs treated with 39 Gy was 153 days, versus 519 days for 5 dogs that received 45 Gy. It appears that radiation therapy prolongs survival times for dogs with brain masses. Although megavoltage therapy would be optimal, orthovoltage radiation can be applied in total doses of 45 Gy in 3.75 Gy fractions over 28 days without adverse effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246211 TI - Hemolytic uremic syndrome in dogs. AB - A disease syndrome similar to the hemolytic uremic syndrome of people is described in three dogs with acute renal failure. In each dog, hemorrhagic gastroenteritis preceded the onset of anuric acute renal failure. Evidence of microangiopathic hemolytic anemia (schizocytes, thrombocytopenia, and increased concentrations of fibrin split products) was present in the three dogs. Serum chemistry results showed increased concentrations of blood urea nitrogen, creatinine, and phosphorus. Ultrasound examination performed in one dog revealed increased echogenicity of the renal cortices. Treatment for anuric acute renal failure using a continuous dopamine and furosemide infusion established urine production in one of three dogs. Microscopic examination of tissue from the two dogs that underwent necropsy showed occlusion of the renal vasculature by fibrin thrombi consistent with microangiopathic arteriolar thrombosis. The pathophysiology and current knowledge of human hemolytic uremic syndrome is compared with hemolytic uremic syndrome in these dogs. PMID- 8246212 TI - The role of vascular mechanisms in the development of acute equine laminitis. PMID- 8246213 TI - Pharmacokinetic and phase I evaluation of carboplatin in dogs. AB - Thirty dogs with spontaneously occurring malignant neoplasms were treated monthly with carboplatin (CBDCA) given as a 30-minute intravenous infusion in a dose escalation study. Twenty-eight dogs were considered evaluable for toxicity. The maximally tolerated dose of CBDCA was conceptually defined as that dose, determined by logistic regression analyses of toxicity data, resulting in a 50% incidence of moderate toxicity (MOD50) or a 5% incidence of severe toxicity (SEV5). Each designated maximally tolerated dose was calculated for the first course of treatment only and for the first and second courses of treatment combined to estimate cumulative drug toxicity. The MOD50 and SEV5 for the first treatment course were 340 and 278 mg/M2, respectively. MOD50 and SEV5 values for the first plus second treatment courses were 327 and 231 mg/M2, respectively. The nadir of neutrophil and platelet counts occurred approximately 14 days after treatment. The mean neutrophil and platelet values for all dogs experiencing myelosuppression during the first two treatment courses were 1541/microL and 62,600/microL, respectively. Nonparametric pharmacokinetic analysis of plasma CBDCA values suggested that half-life (T1/2), area-under-the-curve and total body clearance (CLb) were not dose dependent. Volume of distribution (VDss) significantly increased with dose only between 100 and 150 mg/M2, not between 150 and 300 mg/M2. Dose-independent serum CBDCA pharmacokinetic disposition indicates that detailed investigation of tissue CBDCA distribution would be warranted and may identify novel dosing strategies that could improve the therapeutic index of CBDCA by minimizing toxicity. PMID- 8246214 TI - Urinary indices of horses after intravenous administration of crystalloid solutions. AB - Saline or glucose solution was infused for approximately 4 hours into six healthy mares in two separate experiments to determine the effect of infusion of crystalloid solutions on fractional excretion (FE) of sodium (Na), chloride (Cl), potassium (K), and phosphorus (P), ratio of urinary creatinine to serum creatinine (UCr/SCr), and ratio of urinary osmolality to serum osmolality (Uosm/Sosm). After intravenous infusion of either saline or glucose solution, FENa, FECl and FEP were significantly increased, whereas UCr/SCr and Uosm/Sosm were significantly decreased. In addition, FEK was significantly increased after infusion of glucose solution. It was concluded that urinary indices were altered by intravenous infusion of crystalloid solutions in healthy mares and that fluid therapy may interfere with the use of these indices for diagnostic purposes. PMID- 8246215 TI - Recommendations for standards in transthoracic two-dimensional echocardiography in the dog and cat. Echocardiography Committee of the Specialty of Cardiology, American College of Veterinary Internal Medicine. AB - Recommendations are presented for standardized imaging planes and display conventions for two-dimensional echocardiography in the dog and cat. Three transducer locations ("windows") provide access to consistent imaging planes: the right parasternal location, the left caudal (apical) parasternal location, and the left cranial parasternal location. Recommendations for image display orientations are very similar to those for comparable human cardiac images, with the heart base or cranial aspect of the heart displayed to the examiner's right on the video display. From the right parasternal location, standard views include a long-axis four-chamber view and a long-axis left ventricular outflow view, and short-axis views at the levels of the left ventricular apex, papillary muscles, chordae tendineae, mitral valve, aortic valve, and pulmonary arteries. From the left caudal (apical) location, standard views include long-axis two-chamber and four-chamber views. From the left cranial parasternal location, standard views include a long-axis view of the left ventricular outflow tract and ascending aorta (with variations to image the right atrium and tricuspid valve, and the pulmonary valve and pulmonary artery), and a short-axis view of the aortic root encircled by the right heart. These images are presented by means of idealized line drawings. Adoption of these standards should facilitate consistent performance, recording, teaching, and communicating results of studies obtained by two-dimensional echocardiography. PMID- 8246216 TI - Ultracentrifugal and electrophoretic characteristics of the plasma lipoproteins of miniature schnauzer dogs with idiopathic hyperlipoproteinemia. AB - To better characterize the idiopathic hyperlipoproteinemia of Miniature Schnauzer dogs, the plasma lipoproteins of 20 Miniature Schnauzers (MS) and 11 dogs of other breeds (DOB) were evaluated by ultracentrifugation, electrophoresis, and biochemical tests. Seventeen MS were healthy; 3 had diabetes mellitus. Plasma from 6 of 17 healthy and all 3 diabetic MS was visibly lipemic. Lipemia was slight to marked in healthy lipemic MS, and marked in diabetic ones. All DOB had clear plasma; 8 were healthy and 3 had diabetes. All healthy lipemic MS and diabetic lipemic MS had hypertriglyceridemia associated with excess very low density lipoproteins. Chylomicronemia was present in 4 of 6 healthy lipemic MS and all 3 diabetic lipemic MS. Lipoproteins with ultracentrifugal and electrophoretic characteristics of normal low density lipoprotein were lacking in 4 of 6 healthy lipemic MS. The lipoprotein patterns of 4 of 11 healthy nonlipemic MS were characterized by mild hypertriglyceridemia associated with increased very low density lipoproteins and a lack of lipoproteins with characteristics of normal low density lipoproteins. Lipoprotein patterns of diabetic DOB closely resembled those of healthy DOB; those of diabetic lipemic MS resembled those of markedly lipemic healthy lipemic MS. In conclusion, the hyperlipoproteinemia of Miniature Schnauzers is characterized by increased very low density lipoproteins with or without accompanying chylomicronemia; some affected dogs may have decreased low density lipoproteins. PMID- 8246217 TI - On the mechanism of HRV-14 antiviral compounds: "slow growth" as a conformational search procedure. AB - We report a novel conformational search procedure that is used to investigate the binding mechanism of a member of the WIN class of antiviral compounds. A simple hypothesis of important residues in the binding site based on differences in drug free and drug-bound X-ray structures along with more elaborate models, ultimately including the entire virus, is considered. Our search method is a variant of slow growth molecular dynamics used in free energy simulations and gives rise to local motion in the protein backbone of up to 3 A. This technique involves the scaling of drug-protein interaction energies over time periods of 10-100 ps and gives rise to local motion in the protein backbone. In addition, we have used high temperature dynamics with periodic quenching to generate low-energy conformations with backbone displacements in the crystallographic binding region of up to 7 A from the native structure. Mechanism of binding, hydrogen-bond stabilization of active-site conformations, concerted drug-protein motions, and the mode of virion stabilization are addressed in relation to our ligand induced and high temperature conformational search procedures. A loop-cap like mechanism is consistent with the results of our study. A large movement of the "active-site" residues is shown to be theoretically possible and provides a greater access for entry of the drug into its binding pocket than seen in the available crystal structures. PMID- 8246218 TI - Synthesis and evaluation of 1-acyl-1,2-bis(methylsulfonyl)-2-(2- chloroethyl)hydrazines as antineoplastic agents. AB - A series of 1-acyl-1,2-bis(methylsulfonyl)-2-(2-chloroethyl)hydrazines, conceived as more potent analogs of 1-(2-chloroethyl)-1,2,2-tris(methylsulfonyl)hydrazine, were synthesized and evaluated for antineoplastic activity against the L1210 leukemia in mice. Of these, 1-acetyl-1,2-bis-(methylsulfonyl)-2-(2 chloroethyl)hydrazine produced "cures" of mice bearing the L1210 leukemia at dosage levels that were considerably less than those at which the tris(sulfonyl) analog produced its antineoplastic effects. This compound was also found to have pronounced activity against the P388 leukemia and against several solid tumors, including the B16F10 melanoma, the M5076 reticulum cell sarcoma, and the M109 lung carcinoma. Furthermore, the acyl derivatives were in general considerably more resistant to hydrolysis in aqueous media and more prone to protease- and thiol-mediated activation than the tris(sulfonyl) analog. The former property is important to formulation, while the latter properties may result in some degree of drug targeting and enhancement of the therapeutic indices of these agents. PMID- 8246219 TI - Isosteres of the DNA polymerase inhibitor aphidicolin as potential antiviral agents against human herpes viruses. AB - A variety of isosteres of the DNA polymerase inhibitor aphidicolin were synthesized as potential antiherpes agents. Modeling studies indicated that the bicyclooctane C, D rings of aphidicolin could be replaced by an aromatic moiety while maintaining the spatial arrangement of the hydroxyl group equivalent to the essential C18 hydroxyl group of aphidicolin. Of the racemic isosteres synthesized only 13, the compound with the greatest structural similarity to aphidicolin, showed any significant antiviral activity in primary assays. An enantioselective synthesis of the compound was carried out and the 4aS isomer 36 was shown to account for the observed antiviral activity noted against herpes simplex virus 1 and human cytomegalovirus. PMID- 8246220 TI - Synthesis and CLOGP correlation of imidooxy anticonvulsants. AB - Continuing structure-activity studies on the anticonvulsant activity of analogs of N-(benzyloxy)-2-azaspiro[4.4]nonane-1,3-dione (2a), which displayed anti electroshock seizure (MES) activity and a protective index (TD50/ED50) of > 4.5 are reported. An in-depth analysis of this moiety was studied employing the Topliss structure activity and the Craig plot analytical approaches as well as a semiempirical method. CLOG P analysis was also applied to this series after experimentally determining the NOR fragment. All compounds were minimized and these physicochemical parameters correlated to anticonvulsant activity. Several interesting substituted benzyloxy compounds emerged from this study: the 2',4' dichloro (2b), 4'-(trifluoromethyl) (2c), 2'-bromo (2d), 3'-chloro (2o), 2' chloro (2r), 2'-fluoro (2p), and 3'-fluoro (2w) analogs, all of which had comparable, or better activity than the parent unsubstituted analog (2a). X-ray crystal analysis of the active 2a versus inactive N-benzyl-2-azaspiro[4.4]nonane 1,3-dione (10) is discussed. PMID- 8246221 TI - The synthesis and biochemical pharmacology of enantiomerically pure methylated oxotremorine derivatives. AB - Previous pharmacological studies of methylated oxotremorine derivatives bearing substituents at the 3-, 4-, and 5-positions of the pyrrolidinone ring have been conducted using racemic mixtures, and not with optically active compounds. The synthesis and radioligand binding data of optically active, methylated oxotremorine derivatives at the 3- and 4-positions are described. There are significant pharmacological differences between the 3- and 4-position derivatives. The 4-position enantiomers have weak, approximately equal affinity and antagonist-like profiles, whereas the 3-position enantiomers have significantly different affinities and partial agonist-like profiles. PMID- 8246222 TI - Aminoacyl analogs of chloramphenicol: examination of the kinetics of inhibition of peptide bond formation. AB - Two aminoacyl analogs and one peptidyl analog of chloramphenicol (1, Cl2CHCO-CA) were prepared. These are 2 (L-Phe-CA), 3 (Gly-CA), and 4 (L-Phe-Gly-CA). The kinetics of inhibition of peptide bond formation by these analogs were examined in a cell-free system which was derived from E. coli and used previously for the study of 1 (Drainas; et al. Eur. J. Biochem. 1987, 164, 53-58). In the absence of inhibitor, the reaction follows first-order kinetics for the entire course of the reaction. In the presence of the analog the reaction gives biphasic log-time plots. The kinetic information pertaining to the initial slope of the plot is analyzed (initial-slope analysis). This information differentiates the analogs from the parent compound 1. The parent compound 1 gives complex inhibition kinetics; increasing the concentration of 1 changes the inhibition from competitive to mixed noncompetitive (Drainas; et al. Eur. J. Biochem. 1987, 164, 53-58). In contrast, the analogs give competitive kinetics even at high concentrations of the inhibitor. The following Ki values have been determined: 18.0 microM for 2, 5.5 microM for 3, 1.5 microM for 4. If we were to assume that compounds 2, 3 and 4 behave as classical competitive inhibitors, we could say that 4 is 12 times more potent than 3 and 4 times more potent than 2. On this assumption we could also compare 1 with 4 and see that 4 is 2 times weaker than 1. It is suggested that as compared with 1, the two aminoacyl analogs and the dipeptidyl analog have increased structural similarity to the 3'-terminus of aminoacyl-tRNA or of peptidyl-tRNA and that this similarity results in a more pronounced competitive inhibition. PMID- 8246223 TI - Preparation and anti-HIV activity of O-acylated heparin and dermatan sulfate derivatives with low anticoagulant effect. AB - In order to increase the ratio of anti-HIV activity to anticoagulant activity, glycosaminoglycan derivatives selectively substituted at OH and/or COOH groups were prepared. Standard heparin, heparin fragments, or dermatan sulfate were converted to their tributylammonium or tetrabutylammonium salts. Their selective O-acylation to various (controlled) degrees was carried out in a homogeneous way in N,N-dimethylformamide using carboxylic acid anhydrides and 4 (dimethylamino)pyridine as catalyst. Esterification of the COOH groups was performed by the addition of alkyl halide to an N,N-dimethylformamide solution of glycosaminoglycan tetrabutylammonium salts. The in vitro anticoagulant activity, the activity against HIV-1 and HIV-2 cytopathicity, the cytotoxicity, and the activity on the induction of giant cell formation were determined. O-acylation (O butyrylation or O-hexanoylation) of the heparin fragments obtained by periodate depolymerization (compounds 2d and 2e), and their esters (compounds 7i and 7j), yielded products with very low anticoagulant effects in vitro, yet potent activity against both HIV-1 and HIV-2 induced cytopathicity, and low, if any, cytotoxicity. As compared to other anionic polysaccharides, these acylated derivatives are more active as inhibitors of HIV-induced giant-cell formation. Their anti-HIV activity is related to the degree of O-acylation and is mainly due to the inhibition of virus adsorption to the target cells. PMID- 8246224 TI - A unified framework for using neural networks to build QSARs. AB - We propose a new neural network architecture that explicitly separates linear and nonlinear contributions to the biological activity. To facilitate the use of neural networks as a regular tool we demonstrate that (1) a perceptron with linear output units is equivalent to multiple linear regression and (2) one hidden unit at a time can be added to the network so that QSAR data can be modeled by everything from the simplest linear hypersurfaces to complicated ones. The significant improvements accrued by the use of weight decay are demonstrated. We conclude that models built without attempting weight decay may not be reliable either for interpretation or extrapolation. Finally we compare models generated by neural networks, rank regression, and standard regression on non-normally distributed data and conclude that neural networks like rank regression bring out many facets of the data that are inaccessible to multiple linear regression. All the experiments were done on either triazine inhibition of pure DHFR from L1210 leukemia cells and on the inhibition of intact L1210 leukemia cells sensitive and resistant to methotrexate or on steroid binding to progesterone. PMID- 8246225 TI - Analysis of cocaine receptor site ligand binding by three-dimensional Voronoi site modeling approach. AB - The Voronoi approach has been used to obtain a three-dimensional model for the binding of the cocaine analogues at the cocaine receptor site. The method has been used to determine the geometric details and the physicochemical properties of the binding regions in the receptor site. With only eight compounds in the training set, the Voronoi site model, consisting of four regions, not only fully explains the binding affinity of the input compounds but is also successful in correctly predicting another eight compounds of the test set. The phenyl substituent at the 3-position of the tropane ring of cocaine was found to be the most significant functionality relevant for activity, while moderate contribution results from the hydrophobic interactions of the tropane ring with the binding regions. Some of the problems associated with the approach are discussed, and we report a new procedure for evaluating the validity of the model obtained from our approach. PMID- 8246226 TI - Substituted chromenes as potent, orally active 5-lipoxygenase inhibitors. AB - A series of chromene derivatives was synthesized and evaluated for their in vitro and ex vivo 5-lipoxygenase (5-LO) inhibitory activity. These compounds were prepared by condensation of appropriate salicyl aldehydes with alpha, beta unsaturated carbonyl compounds, followed by transformation to the corresponding hydroxamic acids or N-hydroxyureas. Placement of phenoxy or p-fluorophenoxy substituents at the 6 position of the chromene ring led to a dramatic increase in the in vitro potency as demonstrated by the guinea pig PMN 5-LO assay. Chromene hydroxamic acids, in general, behaved poorly in the ex vivo dog model. On the other hand, replacement of the hydroxamic acid function with N-hydroxyurea yielded potent and long-lasting 5-LO inhibitors in the dog model. In most cases, the oral efficacy of the chromene N-hydroxyureas correlated very well with their in vitro activity. Compounds 43 (CGS 23885) and 55 (CGS 24891) are among the most potent inhibitors prepared, showing IC50 values of 48 and 51 nM, respectively. The values for the duration of action (DA) for compounds 43 and 55 are 21 and 20 h, respectively, following intravenous (i.v.) administration of 1.0 mg/kg. In the oral (po) experiments, 43 and 55 have DA's of 14 and 15 h, respectively, at a 1.0 mg/kg dose. In both iv and po experiments, 43 and 55 showed sustained maximal inhibition (> 95%) at earlier time points. The oral ED50 values of 43 and 55 in the ex vivo dog model are 0.23 and 0.23 mg/kg, respectively, at 6.0 h, and 2.37 and 1.63 mg/kg, respectively, at 24 h. Compound 43, which inhibits sheep seminal vesicle cyclooxygenase (CO) with an IC50 value of 36 microM, was shown to be a selective 5-lipoxygenase inhibitor in the ex vivo study. These compounds compare favorably with zileuton (A-64077) in all the parameters examined. PMID- 8246227 TI - Nonpeptide angiotensin II antagonists derived from 1H-pyrazole-5-carboxylates and 4-aryl-1H-imidazole-5-carboxylates. AB - Two series of potential angiotensin II antagonists derived from carboxyl functionalized "diazole" heterocycles have been prepared and evaluated. Initially, a limited investigation of 4-arylimidazole-5-carboxylates led to 2-n butyl-4-(2-chlorophenyl)-1-[[2'-(1H-tetrazol-5-yl)biphenyl-4-y l] methyl]-1H imidazole-5-carboxylic acid (12b), which was found to be a highly potent antagonist of the rabbit aorta AT1 receptor (IC50 0.55 nM). In conscious, normotensive rats, 12b at 0.1 mg/kg iv inhibited the pressor response to AII by 88%, with a duration of > 6 h. More extensively studied was an isosteric series of 3-alkyl-4-[[2'-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-1H-pyrazole -5- carboxylates bearing aryl, alkyl, or aralkyl substituents at N1. These compounds were available in highly regioselective fashion via condensation of a substituted hydrazine hydrochloride with a 2-(methoxyimino)-4-oxoalkanoate intermediate. In vitro, the most potent pyrazolecarboxylic acids had n-butyl at C3 and were substituted at N1 by such groups as 2,6-dichlorophenyl (19h), 2 (trifluoromethyl)phenyl (19k), benzyl (19t), and phenethyl (19u), all with IC50 values of 0.18-0.24 nM. Although less potent in the receptor assay, 3-n propylpyrazolecarboxylic acids were at least as effective as their butyl counterparts in vivo. Several of the pyrazolecarboxylic acid derivatives demonstrated potent, long-lasting oral activity in rats. At 1 mg/kg po, the 1 benzyl-3-butyl (19t), 1-(2,6-dichlorophenyl)-3-propyl (19v), 3-propyl-1-(2,2,2 trifluoroethyl) (19y), and 1-benzyl-3-propyl (19z) analogues all gave > or = 75% inhibition of the AII pressor response in the rat model, with duration of action > 23 h. PMID- 8246228 TI - Transformation of heterocyclic reversible monoamine oxidase-B inactivators into irreversible inactivators by N-methylation. AB - 3-[4-[(3-Chlorophenyl)methoxy]phenyl]-5-[(methylamino)methyl]- 2-oxazolidinone (1) is a secondary amine known to be a potent time-dependent irreversible inactivator of monoamine oxidase B (MAO-B). The primary amine analogues of derivatives of 1, as well as of the corresponding dihydrofuranone and pyrrolidinone, had been shown to be time-dependent, but reversible, inhibitors of MAO-B. Here it is shown that the primary amine analogue of 1 is a time-dependent reversible inhibitor of MAO-B and that the secondary and tertiary amine analogues of the corresponding oxazolidinone, dihydrofuranone, and pyrrolidinone are time dependent irreversible inhibitors of MAO-B. The reaction leading to the irreversible enzyme adduct formation with 1 can be reversed by increasing the temperature. These results are consistent with a stabilizing stereoelectronic effect on the enzyme adduct caused by N-methylation which hinders free rotation and prevents the sp3-orbital containing the nitrogen nonbonded electrons from being trans to the active site amino acid leaving group. PMID- 8246229 TI - 2-Thio derivatives of dUrd and 5-fluoro-dUrd and their 5'-monophosphates: synthesis, interaction with tumor thymidylate synthase, and in vitro antitumor activity. AB - A convenient synthesis of 5-fluoro-2-thiouracil (11) is based on hydrolytic deamination of 5-fluoro-2-thiocytosine (9). Lewis acid-catalyzed condensation of di-TMS-5-fluoro-2-thiouracil (13) or di-TMS-2-thiouracil (14) with 2-deoxy-3,5-di O-p-toluyl-D-ribofuranosyl chloride (15) led to mixtures of the beta- and alpha anomers of 3',5'-toluylated 2'-deoxy-5-fluoro-2-thiouridine (16 and 18) or 2' deoxy-2-thiouridine (17 and 19), each of which was deblocked with MeOH-NH3 to give the desired free anomeric nucleoside pairs 1, 5 and 3, 7, respectively. These were selectively converted to the corresponding 5'-monophosphates 2, 6 and 4, 8, with the aid of the wheat shoot phosphotransferase system. Conformations of the nucleosides 1, 3, 5, 7 are deduced from 1H NMR spectra, and circular dichroism spectra for nucleotide anomeric pairs 2, 6 and 4, 8 are reported. Whereas beta-2-thio-dUMP (4) was a good substrate (Km approximately 10(-5) M), beta-5-fluoro-2-thio-dUMP (2) proved to be a potent competitive, slow-binding inhibitor (Ki approximately 10(-8) M) of the purified enzymes from Ehrlich ascites carcinoma and L1210 cells. The alpha-anomer 6 was a weak inhibitor, with Ki in the mM range, and its congener 8 hardly interacted with the enzyme. The beta-anomer 1 exhibited antitumor activity in a mouse leukemic cell line L5178Y (IC50 approximately 10(-6) M), hence 40-100-fold weaker than 5-fluoro-dUrd. Its alpha-anomer 5 was 10-fold less active, but exhibited at least 10-fold higher selectivity with respect to the tumor cells than the beta-anomer 1. PMID- 8246230 TI - Synthesis, chemical reactivity, and antitumor evaluation of congeners of carmethizole hydrochloride, an experimental "acylated vinylogous carbinolamine" tumor inhibitor. AB - A series of analogues of 4,5-bis(((N-methylcarbamoyl)oxy)methyl)-1-methyl-2 (methylthio)-im idazole (1, carmethizole) were synthesized. The chemical reactivities of the analogues (as electrophiles) were evaluated and related to the antitumor activity (in vivo and in vitro). Changes in the alkylthio moiety had a significant effect upon the chemical reactivity. Electron-withdrawing groups on the sulfur decreased chemical reactivity and, in parallel, decreased antitumor activity. Carmethizole sulfoxide (11a) was unreactive as an electrophile and exhibited no antitumor activity either in vivo or in vitro; this led to the conclusion that carmethizole sulfoxide was not acting as a "carrier form" of carmethizole. The disulfides 17 and 18 were unreactive as electrophiles but did exhibit antitumor activity. The activity of 17 and 18 was attributed to the thiol 10 that would be generated upon cleavage of the disulfide bond. PMID- 8246231 TI - Synthesis and evaluation of 3-modified 1D-myo-inositols as inhibitors and substrates of phosphatidylinositol synthase and inhibitors of myo-inositol uptake by cells. AB - A number of 3-substituted 1D-myo-inositols were synthesized and evaluated as substrates for phosphatidylinositol synthase and uptake by intact cells. 1D-3 Amino-, -3-chloro-, and -3-(acetylthio)-3-deoxy-myo-inositols were all synthesized by nucleophilic displacement of the 6-O-(trifluoromethyl)sulfonyl group of 1L-1,2:3,4-di-O-cyclohexylidene-5-O-methyl-6-O-[(trifluoromethyl)-sulfon yl] - chiro-inositol (which was prepared from L-quebrachitol), respectively, by reaction with LiN3, followed by reduction of the azido function, and with LiCl and KSAc to give the O-protected compounds. O-Demethylation using BBr3 and concomitant acetal hydrolysis furnished the free-hydroxy 3-amino- and 3-chloro-3 deoxy-1D-myo-inositols. The 3-mercapto analogue was obtained by removal of the acetal groups of the acetylthio analogue, followed by acetylation and purification of the peracetate, and subsequent O-demethylation and deacetylation. The 3-deoxy derivative was synthesized from the 6-O-(imidazol-1-ylthiocarbonyl) compound via Barton-McCombie deoxygenation. The 3-azido derivative was directly synthesized from 1L-1-O-tosyl-chiro-inositol via displacement with azide. The 3 keto analogue was prepared by Pt-catalyzed air oxidation of 1L-chiro-inositol. The compounds were all evaluated as substrates for phosphatidylinositol (PtdIns) synthase from mouse brain. The 3-NH2, 3-F, 3-deoxy, and 3-keto analogues all showed activity as substrates, as measured by liberation of cytidine monophosphate. These compounds also showed inhibition of the reaction of myo [3H]inositol with PtdIns synthase. These results taken together indicate that these compounds are likely to be incorporated into phospholipids. As a further indication that these compounds might be useful as probes for the PtdIns pathway, it was demonstrated that the 3-NH2, 3-F, and 3-deoxy compounds are taken up by intact fibroblast cells as evidenced by their competing with myo-[3H]inositol uptake. PMID- 8246232 TI - Protonation of phosphoramide mustard and other phosphoramides. AB - The chemistry of the bifunctional alkylating agent phosphoramide mustard and model phosphoramides was probed by multinuclear NMR spectroscopy as a function of pH. Between pH 1 and 11, both the 31P and 15N resonances for phosphoramide mustard displayed a single monobasic titration curve with a pKa of 4.9. The protonation below pH 4.9 correlates with the loss in reactivity of the mustard. The 17O NMR spectrum of 17O-enriched phosphoramide mustard shows little change with pH. The data on the mustard was compared to 15N and 31P NMR data on 15N enriched phosphoramidic acid, phosphorodiamidic acid, and phosphoric triamide. Contrary to the conclusions of previous studies, our combined 31P, 15N, and 17O NMR results are more consistent with N-protonation of phosphoramide mustard rather than an O-protonation. Theoretical calculations on the phosphoramidic acid, phosphorodiamidic acid, and phosphoric triamide show O-protonation to be more stable in the gas phase. For the latter two compounds, the calculations suggest that N-protonation may be the most stable protonated form in the aqueous phase. These findings influence our understanding of the structure-activity relationships of phosphoramide mustards. PMID- 8246233 TI - Inhibitors of cholesterol biosynthesis. 1. 3,5-Dihydroxy-7-(N-imidazolyl)-6 heptenoates and -heptanoates, a novel series of HMG-CoA reductase inhibitors. AB - 3,5-Dihydroxy-7-(N-imidazolyl)heptanoates 4 and the corresponding heptenoates 5 were synthesized as novel classes of potent HMG-CoA reductase (HMGR) inhibitors in which members of the latter series possess enzyme inhibitory activity greater than that of lovastatin 1 and pravastatin 2. Structure-activity studies show that the 7-(N-imidazolyl)heptenoates 5 are more active than the corresponding heptanoates 4. For both imidazolyl series, the 4-fluorophenyl group is preferred at C-5, and a broad range of aryl substituents which promote widely different lipophilicities is tolerated at C-4. While the CF3 group is preferred at C-2 in the heptanoate series, the 2-(1-methylethyl) substituent is optimal in the heptenoate series. The 2-(1-methylethyl) and 5-(4-fluorophenyl) groups can be interchanged in the latter series as exemplified by 5ab. Enzyme inhibitory activity resides principally in the 3R,5S series. These potent HMGR inhibitory activities by members of the heptenoate series translated well into whole cell activities in HepG2 cells. X-ray crystallographic studies on the active enantiomer 28 reveal noncoplanarity of the heptenoate C-C double bond with the imidazole ring; this finding provides an explanation for the high acid stability of the heptenoate series. PMID- 8246234 TI - Inhibitors of cholesterol biosynthesis. 2. 3,5-Dihydroxy-7-(N-pyrrolyl)-6 heptenoates, a novel series of HMG-CoA reductase inhibitors. AB - A series of 7-[2,3-diaryl-5-(1-methylethyl)-1H-pyrrol-1-yl]-3,5- dihydroxy-6 heptenoates was prepared and evaluated for its ability to inhibit the enzyme HMG CoA reductase in vitro. Maintaining a 5-(1-methylethyl) substituent found to be optimal in related studies, structure-activity relationships were established for compounds modified at positions 2, 3, and 4 of the pyrrole ring. The 4 fluorophenyl group was preferred at the pyrrole 2-position, while incorporation of a range of substituted phenyl groups and pyridyl substituents at the 3 position provided compounds with equivalent enzyme inhibitory activities and widely different lipophilicities. Pentasubstituted pyrrole 3h was found to have a 10-fold greater potency than lovastatin. PMID- 8246235 TI - Preparation, characterization, cytotoxicity, and mutagenicity of a pair of enantiomeric platinum(II) complexes with the potential to bind enantioselectively to DNA. AB - The synthesis of a pair of enantiomeric Pt(II) complexes, [Pt(R,R-eap)Cl2] and [Pt(S,S-eap)Cl2] (eap = N,N-diethyl-2,4-pentanediamine), designed to bind enantioselectively to GpG and ApG sequences of DNA is described. The in vitro cytotoxicity of each of the enantiomers toward murine leukemia and human bladder tumor cells has been measured. The R,R enantiomer was found to be more active in the leukemia cells, but the difference was not as great as expected (IC50; R,R 14 microM, S,S 33 microM). In the bladder tumor cell line, no significant difference in activity was found. The two enantiomers had similar mutagenicity in the Salmonella reversion assay, but the R,R enantiomer was more cytotoxic in the bacterial cells. A structural analysis of the R,R enantiomer revealed that the ligand adopted an unexpected configuration, and a strain energy minimization analysis showed that this was a consequence of interactions between the diamine ligand and the dichloro ligands. The significance of the structural preferences with respect to the lower than expected enantiospecificity is discussed. Crystals of [Pt(R,R-eap)Cl2] are monoclinic; space group, P2(1)2(1)2(1); a = 7.909(5), b = 12.972(9), and c = 13.269(12) A; Z = 4; and the structure was refined to R = 0.025 (1657F). PMID- 8246236 TI - Possible contribution of a glutathione conjugate to the long-duration action of beta-funaltrexamine. AB - The fumaramate derivative of naltrexone, beta-funaltrexamine (beta-FNA), is a highly selective long-lasting mu opioid receptor antagonist that is active both in vitro and in vivo, presumably as a result of covalent binding to a mu receptor based sulfhydryl group. Glutathione, which occurs in significant levels in brain and liver, was found to undergo a Michael-type reaction with beta-FNA in the test tube to give a stable conjugate 3 which occurred as an isomeric mixture. When tested in the GPI and MVD smooth muscle preparations, 3 was found to possess one tenth the agonist activity of beta-FNA is both tissues, but showed no irreversible antagonist activity. The same result was found for the cysteine conjugate 4, except for some irreversible antagonism in the MVD. Both conjugates antagonize the antinociceptive effect of morphine in the mouse radiant heat tail flick assay on icv administration. This antagonism persisted and actually increased over 24 h and generally paralleled the duration profile of beta-FNA. On sc administration, beta-FNA and 3 showed similar duration of antagonistic effect, while 4 exhibited only marginal activity at the early time interval. When the compounds are compared by the dose to produce equivalent antagonism, beta-FNA and 3 appeared equally effective and accessible by either route, whereas 4 showed a large difference between the two routes. It is possible that the ultra-long antagonism of the conjugates may result from their enzymatic conversion to beta FNA in the central nervous system in view of the fact that conjugate 5, which cannot be converted to beta-FNA, did not produce antagonism of long duration in vivo. Alternatively, the protracted antagonism could arise from sequestration of 3 and 4 in tissue compartments that interface with mu opioid receptors. PMID- 8246237 TI - HMG-CoA reductase inhibitors: design, synthesis, and biological activity of tetrahydroindazole-substituted 3,5-dihydroxy-6-heptenoic acid sodium salts. AB - Compounds comprising a series of 7-[2-(4-fluorophenyl)-4,5,6,7-tetrahydro-2H indazol-3-yl]-3,5- dihydroxy-6-heptenoic acid sodium salts (18) were synthesized and tested for their ability to inhibit HMG-CoA reductase in a partially purified enzyme preparation and cholesterol biosynthesis from acetate in cultured HEP-G2 cells. Changing the size of the saturated ring of the tetrahydroindazole nucleus did not improve potency, but incorporation of substituents at the 7-position resulted in up to 1700-fold improvement in inhibitory potency. Structure-activity studies revealed that the most potent compounds possess a substituted benzyl group at the 7-position, with a preference for steric bulk at the para position of the benzene ring. The most potent enzyme inhibitor (18t, IC50 = 3.0 nM) is approximately 3-fold more potent than lovastin sodium salt (2). The most potent cholesterol biosynthesis inhibitor in HEP-G2 cells (18q, IC50 = 0.078 microM) is slightly less potent than 2 (sodium salt). Molecular modeling studies suggested that, when compared to the parent compound (18b) lacking the appropriate 7 substituent, 18t overlaps better with 2 and literature inhibitors 5 and 6 in a hydrophobic binding region adjacent to the enzyme active site. PMID- 8246238 TI - Synthesis and evaluation of new 6-amino-substituted benzo[c]phenanthridine derivatives. AB - Different 7,8,9,10-tetrahydrobenzo[c]phenanthridin-6(5H)-ones (10a-e) were prepared by using a one-pot procedure which includes the preparation of various 6 and 7-alkoxy-1-naphthylisocyanates from 1-naphthylamines and triphosgene, followed by addition of 1-N-morpholino-1-cyclohexenes, and cyclization of the resulting amides upon heating in the presence of hydrogen chloride. Subsequent aromatization, chlorination, and substitution with (dimethylamino)alkylamines, followed by a demethylation or a selective desisopropylation, allowed us to synthesize the derivatives 6a-i and 7a-h bearing a [(dimethylamino)alkyl]amino side chain at their 6-position. These compounds, as the other analogs 5a-b, were devised to further study the structure-activity relationships in the benzo[c]phenanthridine family of antitumor alkaloids led by fagaronine (1a) and nitidine (1b). Topoisomerases I and II cleavable complex assay and evaluation of the cytotoxicity and antitumor properties were performed. In vitro cytotoxicity (L1210 and Calc 18) shows a relationship between the cytotoxicity of these compounds and their topoisomerase poisoning properties. However, all these compounds were devoid of significant antitumor effect on the P388 murine leukemia system. PMID- 8246239 TI - Development of high-affinity 5-HT3 receptor antagonists. Structure-affinity relationships of novel 1,7-annelated indole derivatives. AB - On the basis of the structures of ondansetron and GR 65,630, its ring-opened C linked methylimidazole analogue, novel 1,7-annelated indole derivatives were synthesized as potential 5-HT3 antagonists. Receptor binding studies show that all compounds display a high affinity for the 5-HT3 receptors. In both series annelation results in compounds being 7 and 4 times more potent than the references ondansetron and GR 65,630, respectively. Similar to ondansetron, the 1,7-annelated indoles show little stereoselectivity. The (-)-isomers are only slightly more potent than the (+)-isomers. The receptor binding profile of l-10 [(2-methyl-1H-imidazol-1-yl)methyl]-5,6,8,9,10,11-hexahydro-4H-pyri do [3,2,1 jk]carbazol-11-one hydrochloride (24b) (INN cilansetron) shows that the compound displays, besides a high affinity for 5-HT3 receptors (Ki = 0.19 nM), a weak affinity for sigma-receptors (Ki = 340 nM), muscarine M1 receptors (Ki = 910 nM), and 5-HT4 receptors (Ki = 960 nM) and no affinity (Ki > or = 5000 nM) for all the other receptor types tested (n = 37). The new compounds fit the proposed necessary chemical template for binding: a heteroaromatic ring system, a coplanar carbonyl group, and a nitrogen center at well-defined distances. The enhanced potency of the annelated 1,7-indole derivatives indicates that the extra ring provides a favorable hydrophobic area for interaction with the 5-HT3 receptor site. In vivo cilansetron is more potent and induces less central side effects than ondansetron. At present cilansetron is in clinical trials. PMID- 8246240 TI - Synthesis and pharmacological examination of benzofuran, indan, and tetralin analogues of 3,4-(methylenedioxy)amphetamine. AB - Benzofuran, indan and tetrahydronaphthalene analogs of 3,4 (methylenedioxy)amphetamine (MDA) were prepared in order to examine the role of the dioxole ring oxygen atoms of MDA in interacting with the serotonin and catecholamine uptake carriers. The series of compounds was evaluated for discriminative stimulus effects in rats trained to discriminate saline from the training drugs (S)-(+)-MBDB (1c), MMAI (3), and (S)-(+)-amphetamine and for the ability to inhibit the uptake of [3H]serotonin, [3H]dopamine, and [3H]norepinephrine into crude synaptosome preparations. Behaviorally, the benzofuran and indan analogs 4-6 produced similar discriminative cues, whereas the tetralin derivative 7 did not fully substitute for the training drugs. The results in the in vitro pharmacology studies indicate that selectivity for 5-HT versus catecholamine uptake carriers may be modulated by the position and orientation of ring oxygen atoms. However, the nonoxygenated isostere 6 possessed high potency at all uptake sites examined. Enlargement of the saturated ring by one methylene unit to give the tetralin derivative resulted in a large (3-4-fold) reduction in activity at catecholamine sites. PMID- 8246241 TI - 18F-labeled benzamides for studying the dopamine D2 receptor with positron emission tomography. AB - Two series of (N-benzylpiperidin-4-yl)- and (9-azabicyclo[3.3.1]nonan- 3 beta yl)benzamides were prepared, and in vitro binding assays were used to measure the affinity of these compounds for dopamine D2, dopamine D3, serotonin 5-HT2, and alpha 2-adrenergic receptors. The results of these studies indicated compounds 23, 26b, and 34 have the selectivity needed for in vivo studies of the D2 (and possibly D3) receptors. 18F-Labeled analogues of 23, 26b and 34 were prepared by N-alkylation of the corresponding desbenzyl precursors with [18F]-4-fluorobenzyl iodide. Preliminary in vivo studies demonstrated that [18F]-23 and [18F]-26b are suitable candidates for further evaluation in positron emission tomography imaging studies. The slow rate of washout of [18F]-34 from nondopaminergic regions and its comparatively high lipophilicity indicates that this compound may not be suitable for imaging studies because of a high level of nonspecific binding. PMID- 8246242 TI - Synthesis and antiviral activity of 5'-deoxypyrazofurin. AB - In searching for derivatives of pyrazofurin that could display antiviral properties by means that do not require C-5' phosphorylation, 5'-deoxypyrazofurin (3) has been synthesized in six steps from methyl5-deoxy-2,3-O-isopropylidene beta-D-ribofuranoside (4). Compound 3 was evaluated for antiviral activity against a large number of viruses including herpes-, pox-, myxo-, toga-, arena-, rhabdo-, picorna-,reo-, and retroviruses. Compound 3 proved active against respiratory syncytial virus (in HeLa cells), vaccinia virus (in embryonic skin muscle fibroblast cells), vesicular stomatitis virus (in HeLa cells), and influenza A virus (in Madin-Darby canine kidney cells) at concentrations (ranging from 4 to 20 micrograms/mL) that were nontoxic to the confluent host cell cultures. PMID- 8246243 TI - Photoisomerization of a potent and selective adenosine A2 antagonist, (E)-1,3 Dipropyl-8-(3,4-dimethoxystyryl)-7-methylxanthine. PMID- 8246244 TI - Synthesis and biological activity of some methoctramine-related tetraamines bearing a 11-acetyl-5,11-dihydro-6H-pyrido[2,3-b][1,4]-benzodiazepin-6-one moiety as antimuscarinics: a second generation of highly selective M2 muscarinic receptor antagonists. PMID- 8246245 TI - (Dipropylphenoxy)phenylacetic acids: a new generation of nonpeptide angiotensin II receptor antagonists. PMID- 8246246 TI - A homologous series of permanently charged 1,4-dihydropyridines: novel probes designed to localize drug binding sites on ion channels. PMID- 8246247 TI - Hepatitis C virus infection--prognosis and treatment. PMID- 8246248 TI - Typing of Aeromonas spp. by numerical analysis of immunoblotted SDS-PAGE gels. AB - One hundred and three isolates of Aeromonas spp., collected from both environmental sources and patients, were examined by SDS-PAGE of whole cells followed by immunoblotting with polyclonal rabbit antiserum raised against whole cells of A. sobria. All isolates were typable, yielding 15-20 well separated bands. Reproducibility of the technique was good and discrimination excellent, yielding 30 types amongst 103 isolates. Immunoblot type was not related to biochemical phenotype. Attempts to correlate immunoblot type with serotype were unsuccessful because only 42% of the strains tested could be serotyped. PMID- 8246249 TI - Determination of the optimal transport system for Helicobacter pylori cultures. AB - A range of solid and liquid media was evaluated for the ability to maintain survival of Helicobacter pylori strains under different conditions. Chocolate agar slopes maintained survival of most strains for longer than 3 days, some strains surviving for up to 9 days, despite a decreased number of viable cells. Temperature and atmosphere did not significantly influence the performance of these slopes. The BBL Campy Pouch system also achieved a considerable recovery rate of H. pylori after storage for 3 days at the same range of temperatures. Brain-heart infusion broth with horse serum was superior among the liquid media tested, maintaining the viability of H. pylori for c. 3 days at temperatures ranging from -4 degrees C to 21 degrees C. Chocolate agar slopes are recommended as suitable for transport of H. pylori strains. PMID- 8246250 TI - Evaluation of the polymerase chain reaction for detecting the urease C gene of Helicobacter pylori in gastric biopsy samples and dental plaque. AB - A polymerase chain reaction (PCR) assay with oligonucleotide primers homologous to a portion of the urease C gene of Helicobacter pylori was evaluated for specificity with pure DNA and biopsy material. The assay was used to test for the presence of the organism in dental plaque. The species specificity of detection was confirmed by ensuring that the primers did not amplify DNA extracts from H. cinaedi, H. felis, H. fennelliae, H. mustelae and H. nemestrinae. Sixty-two gastric biopsy samples collected from 14 patients (antrum, body and duodenal sites) were cultured and PCR was performed on the samples after culture. Primer sites were conserved in genomically diverse strains. Samples prepared by single step heat lysis of bacterial cells and biopsy material did not inhibit PCR. The overall specificity was 96% irrespective of genotype. H. pylori was not cultured from dental plaque (15 patients), neither was H. pylori DNA detected by PCR in either urea breath test-positive or -negative individuals. The results showed that primer pair sequences within the urease C gene are conserved in most strains and provide an accurate basis for detecting H. pylori. As the PCR assay was not inhibited and did not yield false positive results with crude extracts from organisms or in the presence of biopsy material, its value as a diagnostic test was confirmed. PMID- 8246251 TI - An investigation of a nosocomial outbreak of Clostridium difficile by pyrolysis mass spectrometry. AB - Isolates from a presumptive nosocomial outbreak of Clostridium difficile infection at a large teaching hospital were typed by pyrolysis mass spectrometry (PMS) and antibiograms. One isolate, from the putative index case, was dissimilar from the outbreak strain, but 24 isolates from 16 patients were indistinguishable by both methods. The outbreak centred on two wards for the acute care of the elderly, with a few cases elsewhere. Transfer of patients appeared to be the route of transmission between wards. There was a significant fall in the incidence of cases following intervention by the Infection Control Unit. This included ward inspection, advice on antibiotic usage and advice on prevention of faecal-oral transfer, particularly by proper handwashing. Subsequent monitoring of C. difficile infection showed a background of sporadic, dissimilar isolates with occasional apparent cross-infection incidents limited to a few patients. In suspected outbreaks, patterns of antibiotic susceptibility may be useful in initial screening, before referral for more sophisticated typing. There was excellent correlation between PMS results, antibiograms and epidemiological information. PMID- 8246252 TI - Biologically active extracellular products of oral viridans streptococci and the aetiology of Kawasaki disease. AB - A bacteriological study of isolates from the oral cavity of patients with Kawasaki disease (KD), age-matched non-KD patients and healthy children, showed that over half the KD and control isolates had gram-positive, catalase-negative cocci. About 50% of these organisms were identified as viridans streptococci by means of an API Strep 20 kit. Further identification by fluorometric DNA-DNA hybridisation demonstrated that the predominant species were S. oralis and S. mitis, each of which accounted for 25% of the isolates of viridans streptococci; 40% of viridans strains were unidentifiable; and S. sanguis and S. parasanguis were minor components. Studies in vivo showed that insertion of culture supernates of most of the viridans streptococci increased capillary permeability and induced redness with swelling and occasional bleeding in rabbit skin. One third of S. mitis strains and one-fifth of the unidentified strains caused aggregation of human blood platelets, whereas S. oralis and other strains had no such effect. The distribution of extracellular lipoteichoic acids and glucan produced in the presence of sucrose was also examined. There were no significant differences in the recovery rate of viridans streptococci forming these biologically active extracellular products between KD and control groups. PMID- 8246253 TI - Simultaneous expression of smooth and rough phase properties related to lipopolysaccharide in a strain of Brucella melitensis. AB - Brucella strains exhibit either a rough (R) or a smooth (S) colonial phase identifiable by bacteriological methods. This depends on the biosynthesis and translocation to the surface in S but not in R strains, of the O-polysaccharide chain of the lipopolysaccharide (LPS) molecule. B. melitensis biovar 1 strain EP exhibited simultaneously both S and R characteristics in relation to colonial morphology, agglutination by monospecific anti-M and anti-R sera, activity of bacteriophages lytic for rough Brucella spp. (phage R/C) and for smooth B. melitensis (phage Iz). B. melitensis strain EP expressed fewer O-chains with a similar distribution of molecular weights than B. melitensis reference strain 16M by SDS-PAGE and immunoblotting, but higher amounts of R-LPS. Quantitative determination of S-LPS by a turbidimetric latex inhibition immunoassay with monoclonal antibodies confirmed the limited expression of S-LPS in strain EP. As with other gram-negative bacteria, the phenomenon could be attributed to a deficiency in one step of the biosynthetic assembly of the O-chains. PMID- 8246254 TI - Further studies of clinical isolates of cysteine-requiring Escherichia coli and Klebsiella and possible mechanisms for their selection in vivo. AB - Cysteine-dependent (cys-) Escherichia coli and Klebsiella spp., defective in sulphate assimilation, were isolated from urine and stool samples of infected patients. These isolates reverted to prototrophy under conditions of cysteine deprivation but the revertant strains and a prototrophic wild-type E. coli strain became auxotrophic for cysteine in a cysteine-enriched medium. This suggested that excess cysteine acts as a repressor of the cys HIJ operon known to control aspects of cysteine biosynthesis. A group of mostly elderly patients infected with cys- strains suffered a disproportionate amount of renal impairment as compared with a control group. In renal impairment, sulphur compounds, including cysteine, are retained. This raises the possibility that these raised levels of cysteine and related compounds may enhance the selection of cys- strains in vivo. PMID- 8246255 TI - Purification and characterisation of haemagglutinin from Bordetella bronchiseptica. AB - A surface protein of Bordetella bronchiseptica was purified in one step by affinity chromatography with bovine submaxillary mucin coupled to agarose. The purified protein, with a mol. wt of 200 kDa and an iso-electric point of pI 6.5, showed haemagglutinating activity for bovine erythrocytes. This haemagglutinin (HA) inhibited the adherence of B. bronchiseptica to a rat lung cell line (L2) and was able to bind to N-acetylneuraminic acid. These findings suggest that the HA of B. bronchiseptica is an adhesin. PMID- 8246256 TI - Identification of the 33-kDa alkaline protease of Aspergillus fumigatus in vitro and in vivo. AB - Aspergillus fumigatus produced a 33-kDa serine protease (ALP) in vitro and in vivo. In vitro, this alkaline protease was secreted when the fungus was cultivated in the absence of protein, if the pH of the medium remained close to neutrality. Western blotting and immunoelectronmicroscopy studies showed that ALP was localised in the wall of the fungus and was degraded after secretion in the culture medium under conditions of low pH. Although present in the lung during infection, ALP did not appear to be diagnostically useful and was different from the precipitating chymotrypsin antigen used in the diagnosis of aspergilloma. PMID- 8246257 TI - Forsaking the tome--a worms' eye view of taxonomy. PMID- 8246258 TI - Detection of serum and faecal antibodies in haemorrhagic colitis caused by Escherichia coli O157. AB - Cases of culture-confirmed clinically typical haemorrhagic colitis caused by verocytotoxin-producing (VT+) Escherichia coli O157 and age- and sex-matched control patients were examined for antibodies to E. coli O157. Serum samples from 28 cases and 34 patients in control group 1 were examined for VT1- and VT2 neutralising antibodies, E. coli O157 agglutinating antibodies, and by an enzyme immunoassay (EIA) technique for IgG antibodies against smooth lipopolysaccharide purified from E. coli O157 and for IgG antibodies against whole intact E. coli O157 cells. Differences between antibody titres were significant when compared by a Wilcoxon two-sample test for E. coli O157 agglutinating antibodies (p < 0.05) and IgG antibodies against whole cells (p < 0.001). The whole-cell EIA was used further to examine faecal samples from 93 cases and 47 patients in control group 2 for IgA antibodies. Elevated levels of faecal IgA specific for E. coli O157 were found in 59 (63.4%) of 93 cases but in only 10 (21.2%) of 47 control patients (p < 0.001); follow-up faecal samples from five cases all showed marked rises in levels of IgA that appeared to coincide with cessation of excretion of the organism. Detection of specific faecal IgA with a whole-cell EIA, although requiring further evaluation, may be a useful addition to tests currently available for the diagnosis of infection by VT+ E. coli O157. PMID- 8246259 TI - Comparison of three molecular typing methods in studies of transmission of Porphyromonas gingivalis. AB - Porphyromonas gingivalis is associated strongly with severe periodontitis, but little information is available on possible transmission routes of this species. This study evaluated three DNA-based molecular typing methods for use in epidemiological surveys of P. gingivalis. In total, 32 isolates from eight married couples were investigated by: (i) restriction endonuclease analysis (REA) of whole chromosomal DNA; (ii) hybridisation of DNA fragments with ribosomal DNA (ribotyping); and (iii) amplification of DNA by the polymerase chain reaction with arbitrary primers (AP-PCR). The data obtained with the three methods were in broad agreement: in six of the eight couples, the isolates from husband and wife were indistinguishable, but isolates from unrelated individuals showed distinct types with all three methods. For some isolates, minor differences in REA pattern were obtained which could not be correlated with differences in ribotype or AP PCR type. Ribotyping showed differences between isolates from one individual, which were indistinguishable with the other two methods. The patterns obtained with ribotyping or AP-PCR were simple in comparison to the relatively complex REA patterns. Although all three methods were concordant, AP-PCR was found to be the least time-consuming method. The data support the suggestion that P. gingivalis can be transmitted between spouses. PMID- 8246260 TI - Co-elimination of mec and spa genes in Staphylococcus aureus and the effect of agr and protein A production on bacterial adherence to cell monolayers. AB - Phenotypic loss of protein A production was tested in six methicillin-resistant (McR) Staphylococcus aureus (MRSA) isolates and their isogenic methicillin sensitive (McS) variants by a radiolabelled IgG-binding assay with washed cells and by Western blotting of supernates prepared from lysed washed cells. Genomic DNA was probed for homology with the protein A gene (spa) in EcoRI digests and for homology to the methicillin resistance gene (mec) in HindIII digests. The McS variants had lost homology with mec. An isogenic pair of McR and McS strains, and derivatives of S. aureus 8325-4 with site-specific mutations of the accessory gene regulator locus (agr) and spa, were tested for adherence to human peritoneal mesothelial cells in monolayer culture. The isogenic pair were also tested for adherence to HEp-2 and Vero cell monolayers in assays with 3H thymidine-labelled bacteria. McR isolates produced protein A which was absent from three strains that had become McS. This correlated with deletion of the spa locus. Spa homology, but reduced production of protein A, was retained in one McS strain which also showed reduced adherence to HEp-2, Vero and mesothelial cells (p < 0.05) compared with the parent McR strain. A spa mutation in strain 8325-4 did not significantly affect adherence to mesothelial cells but mutation in agr increased adherence significantly in both Spa+ and Spa- strains. PMID- 8246261 TI - Evaluation of the capacity of oral streptococci to produce hydrogen peroxide. AB - The capacity of 11 strains of oral streptococcal species (Streptococcus sanguis, S. oralis, S. mitis and S. sobrinus) to produce hydrogen peroxide (H2O2) was studied in vitro. Detection of this property in solid media, particularly with trypticase soy agar-benzidine-peroxidase, was more sensitive than in liquid media. The addition of carbohydrates (arabinose, xylose, mannose, sorbose and lactose), sorbitol and saccharine to buffered trypticase soy broth increased H2O2 production in S. oralis NCTC 11427, although the concentrations obtained with some substrates (glucose, galactose, mannitol and xylitol) were lower than those obtained in controls. In S. sanguis NCTC 7863, H2O2 production was detected only with galactose, sorbitol, lactose and saccharin. PMID- 8246262 TI - The agglutination of beta-haemolytic streptococci by lectins. AB - The ability of 25 lectins, isolated from different plants and fungi, to agglutinate 95 clinical isolates of beta-haemolytic streptococci was examined. Cell suspensions were untreated, trypsin-treated or boiled at pH 2.0. None of the 95 untreated cell suspensions gave a visible reaction with any of the lectins. When the cells were trypsinised, 42 strains were agglutinated with one or more lectin and after boiling at pH 2, all the strains were agglutinated. After treatment with trypsin, 20 different agglutination patterns were observed, and after boiling, 19 patterns, four of which were similar. A correlation was found between Lancefield group C and some of these patterns. Some lectins reacted specifically with group C streptococci; DBA and WFA, both specific for D-GalNAc, DSA, a GlcNAc-specific lectin, and RPA, which showed a complex specificity, reacted only with group C strains. Furthermore, the lectin of Maackia amurensis reacted with 50% of group B streptococci only. Agglutination assays with lectins were reproducible, easy to perform, relatively inexpensive and, therefore, applicable to studies of cell-wall structure and epidemiology of beta-haemolytic streptococci. PMID- 8246263 TI - Probes for the study of mupirocin resistance in staphylococci. AB - Probes constructed from a 4.05-kb EcoRI digest fragment of a mupirocin resistance plasmid and a 751-bp internal part of this fragment hybridised with DNA from all of 36 independent high-level mupirocin-resistant staphylococci tested from seven centres; most were Staphylococcus aureus. In most instances the probes detected an EcoRI digest fragment of approximately 4 kb. Probes did not hybridise to DNA from low-level resistant strains, nor from strains sensitive to mupirocin. PMID- 8246264 TI - Mupirocin resistance in coagulase-negative staphylococci. AB - High-level mupirocin resistance in coagulase-negative staphylococci isolated from patients undergoing peritoneal dialysis was investigated by transfer of the resistance determinants, usually in the form of a plasmid, to Staphylococcus aureus strains, cleavage of the plasmid by restriction endonuclease and hybridisation with a probe comprising a 4.05 kb EcoRI fragment of a plasmid from a S. aureus strain. In most instances the mupirocin-resistant staphylococci isolated from each patient were different according to the species, antibiogram and plasmid profile data. The mupirocin resistance determinant was carried on various plasmids as judged by EcoRI restriction fragment length polymorphisms. All hybridised at about 4 kb with the S. aureus probe. PMID- 8246265 TI - Insertion sequence IS200 can differentiate drug-resistant and drug-sensitive Salmonella typhi of Vi-phage types E1 and M1. AB - The type strains of Vi-phage types E1, M1 and A of Salmonella typhi, together with drug-resistant and drug-sensitive strains of phage types E1 and M1 isolated in 1992 from patients associated with India or Pakistan, and a drug-resistant strain of phage type A isolated in South Africa in 1991, were characterised with respect to the presence of plasmids conferring resistance to antimicrobial drugs and their chromosomal insertion sequence IS200 profiles. The three type strains, the drug-sensitive strains of Vi-phage types E1 and M1, and a strain of phage type M1 resistant to ampicillin and trimethoprim but not to chloramphenicol, did not contain plasmids. In contrast, for strains of phage types E1 and M1 resistant to chloramphenicol, ampicillin and trimethoprim, and for the drug-resistant strain of phage type A, the complete spectrum of resistance was encoded by high molecular mass plasmids belonging to the H1 incompatibility group. Characterisation of IS200 profiles demonstrated that at least 13 IS200 copies were distributed on the chromosome of all strains tested. Although the IS200 profiles of the type strains of Vi-phage types A, E1 and M1 were identical, it was possible to distinguish between drug-sensitive and drug-resistant strains of Vi-phage types E1 and M1 isolated from patients infected in India and Pakistan by this method. It was concluded that although IS200 typing is not as discriminatory as phage typing for the primary subdivision of S. typhi, it may be useful for certain epidemiological investigations and, in particular, for investigating the origins of strains with multiple drug resistance. PMID- 8246266 TI - Inhibition of the adherence of Pseudomonas aeruginosa to epithelial cells by IgG subclass antibodies. AB - A beneficial role of the antibody response to Pseudomonas aeruginosa seen in cystic fibrosis (CF) patients has not been established. We investigated a possible role for these antibodies as inhibitors of the adherence of P. aeruginosa to mammalian cells. An adhesion model system was used, employing buccal epithelial cells in an enzyme-labelled immunoassay procedure on microtitration plates. Total levels of IgG, IgA, IgM and the four IgG subclasses were estimated in 11 CF patients and 10 healthy controls. Most of the CF patients demonstrated increased levels of all these immunoglobulin types. Sera from seven patients with elevated serum IgG were observed to cause greater inhibition of the adherence of P. aeruginosa to buccal cells than were the sera from four CF patients with low serum IgG and from ten healthy controls. Nevertheless, the levels of individual anti-P. aeruginosa IgG subclass antibodies varied amongst the patients and did not correlate with the degree of inhibition of bacterial adherence. Negative affinity chromatography was used to obtain antibody fractions enriched for IgG1, IgG2 or IgG4 and protein A-sepharose chromatography was used to isolate IgG3 antibodies from CF patients. The IgG1-, IgG2-, or IgG4-enriched fractions similarly inhibited the adherence of P. aeruginosa in the test system, whereas three of five IgG3-enriched fractions from CF patients had no greater effect on adhesion than did IgG from control individuals. PMID- 8246267 TI - Role of IgG subclass response to outer-membrane proteins in inhibiting adhesion of Pseudomonas aeruginosa to epithelial cells. AB - The IgG subclass response to the major outer-membrane proteins (OMPs) of Pseudomonas aeruginosa was investigated in 11 cystic fibrosis (CF) patients and 10 healthy controls. Inhibition of adhesion of P. aeruginosa to buccal epithelial cells by the IgG serum fractions from the CF patients has been established previously. The CF patients demonstrated marked heterogeneity in their individual IgG subclass response to pseudomonal OMPs. The predominant IgG1 and IgG4 responses were directed towards OMPs F, H2 and, with IgG1 only, to protein I. Proteins of 42 and 46 kDa primarily elicited an IgG2 response but some patients produced IgG4 antibodies. The IgG3 response varied from very weak in some patients to a strong reaction with proteins D2, E, G and I in others. The range of antigen-specific IgG subclass responses was similar in CF patients whose IgG fractions strongly inhibited the adherence of P. aeruginosa to epithelial cells and in those whose fractions gave only weak inhibition of adherence. There was no indication that an antibody response towards any particular OMP was implicated in the inhibition of bacterial adherence. Thus, the IgG subclass response to OMPs did not exert a significant effect on adherence when investigated in isolation, but may possibly play some role in combination with other processes. PMID- 8246268 TI - Serum hepatitis B virus DNA detection with S- and C-region-directed probes. AB - Developments in molecular biology have offered a wide range of nucleic acid probes to detect the genome of hepatitis B virus (HBV). We have tested the ability of two enzyme-linked (alkaline phosphatase) probes to detect HBV-DNA. These hybridise with the S and C regions of the genome of HBV and are used to determine the clinical significance of detecting the two regions. A total of 66 serum samples from patients at different stages of HBV infection was examined. HBV-DNA was detected with at least one of the probes in 17 (85%) patients with HBeAg-positive chronic hepatitis, five (26.3%) with anti-HBe-positive chronic hepatitis and six (66.6%) with acute hepatitis. Although both probes were able to detect as little as 10 pg/ml (2.86 x 10(6) g.E./ml) of a full length HBV-DNA standard, the C-region-directed probe did not react in one patient with acute hepatitis, two with HBeAg-positive and three with anti-HBe-positive chronic hepatitis. When C-region-directed probes are used for diagnostic purposes, results should always be accompanied by hybridisation with probes directed against other regions showing less variability (e.g. S region). PMID- 8246269 TI - Promises and problems of biological atomic force microscopy. PMID- 8246270 TI - A new method for investigating the undersurface of cell monolayers by scanning electron microscopy. AB - Glow discharge is commonly used for cleaning the inside of coating units and for cleaning hard surfaces before carbon or metal evaporation procedures. In this study it has been used to remove the embedding medium to reveal, for scanning electron microscope (SEM) study, the undersurfaces of Balb/c 3T3 fibroblastic cells that have been cultured on Thermanox discs and embedded in LR White resin. Ten to twenty-minute ionization times were found to reveal the largest area of the undersurface without causing damage to the cells. Chemical etching of the resin was also shown to reveal the undersurface of the cells, but caused some damage, preventing successful re-embedding for transmission electron microscopy, and at higher magnifications revealed less detail. A circular impression within the main outline of the cells was observed in many cells, which is considered to reflect the presence of a nucleus. The undersurfaces of most cells, after applying both methods of etching, displayed a number of very short processes. Subsequent transmission electron microscopy of ultrathin sectioned, re-embedded, areas of the gold sputter-coated blocks revealed the depth of ionization that had occurred and confirmed that the specimens observed in SEM were the undersurfaces of cells. This method can be modified to study the attaching surface of any organism to a substratum. PMID- 8246271 TI - Standards for quantification of elements in the otolithic membrane by electron probe X-ray microanalysis: calibration curves and electron beam sensitivity. AB - An absolute quantitative standardization technique has been developed to measure Ca and K weight fractions (WF) in the otolithic membrane of the saccule and utricle by scanning electron microscopy and electron probe X-ray analysis using the peak-to-background (P/B) ratio method. Microcrystalline salt standards were used to calibrate Ca and K K alpha P/B or Y = (P/B).Z2/A (Z = atomic number; A = atomic weight) against WF at 10, 15, 20 and 25 kV accelerating voltage. The effect of voltage on the calibration, plotting the coefficient of correlation (r) as a function of voltage, was not dependent on the voltage in the range 10-25 kV for Ca standards. K standards were also independent when P/B was corrected for Z2/A. Background counts in the otoconia (Bo) were obtained at 5, 25, 50, 100, 200 and 500 s and used to test the electron beam sensitivity of saccular and utricular otoconia. Bo was not dependent on the spectra acquisition time, with the exception of Bo under K alpha K peak in the saccule at 10 kV. Ca and K WF were determined at 10, 15, 20 and 25 kV in the saccule and utricle, showing similar values regardless of the voltage used. This method of calibration offers several advantages, such as stability, homogeneity, known composition of the standards, high reproducibility at different voltages even without Z2/A correction and the similarity between the otoconia and crystal standards. We recommend the application of this method for other elements and biomineral systems. PMID- 8246272 TI - Unbiased estimation of human body composition by the Cavalieri method using magnetic resonance imaging. AB - The classical methods for estimating the volume of human body compartments in vivo (e.g. skin-fold thickness for fat, radioisotope counting for different compartments, etc.) are generally indirect and rely on essentially empirical relationships--hence they are biased to unknown degrees. The advent of modern non invasive scanning techniques, such as X-ray computed tomography (CT) and magnetic resonance imaging (MRI) is now widening the scope of volume quantification, especially in combination with stereological methods. Apart from its superior soft tissue contrast, MRI enjoys the distinct advantage of not using ionizing radiations. By a proper landmarking and control of the scanner couch, an adult male volunteer was scanned exhaustively into parallel systematic MR 'sections'. Four compartments were defined, namely bone, muscle, organs and fat (which included the skin), and their corresponding volumes were easily and efficiently estimated by the Cavalieri method: the total section area of a compartment times the section interval estimates the volume of the compartment without bias. Formulae and nomograms are given to predict the errors and to optimize the design. To estimate an individual's muscle volume with a 5% coefficient of error, 10 sections and less than 10 min point counting (to estimate the relevant section areas) are required. Bone and fat require about twice as much work. To estimate the mean muscle volume of a population with the same error contribution, from a random sample of six subjects, the workload per subject can be divided by square root of 6, namely 4 min per subject. For a given number of sections planimetry would be as accurate but far more time consuming than point counting. PMID- 8246273 TI - Prevalence of sexual dysfunction in women: results of a survey study of 329 women in an outpatient gynecological clinic. AB - Few studies have investigated the prevalence of sexual dysfunction in nonclinical samples. In the present study, a standardized sexual function questionnaire was administered to 329 healthy women, aged 18-73 years, all of whom were enrolled in a Women's Wellness Center. About two-thirds of the sample were married or living with a partner, and most women were employed outside of the home. A broad range of sexual behavior frequencies were observed, with 48.5% reporting at least weekly intercourse, compared to 28.4% who were not sexually active at the time of study. Among the most common sexual problems reported were anxiety or inhibition during sexual activity (38.1%), lack of sexual pleasure (16.3%), and difficulty in achieving orgasm (15.4%). Other common problems were lack of lubrication (13.6%) and painful intercourse (11.3%), each of which was significantly more prevalent in the postmenopausal group. Despite these difficulties, 68.6% of the sample rated their overall sexual relationship as satisfactory. Age and relationship status were significant predictors of sexual satisfaction, with older women and singles reporting a higher incidence of sexual problems. Educational level, religious affiliation, and employment status were not predictive of sexual dysfunction in the present study. PMID- 8246274 TI - Penile sensitivity in men with premature ejaculation and erectile dysfunction. AB - Previous research indicates that penile sensitivity is typically lower in men with erectile dysfunction than in age-matched controls. On the assumption that sensitivity might be greater in men with short ejaculation latency (premature ejaculation), the present research investigated penile threshold (sensitivity) to vibrotactile stimulation in men with premature ejaculation, erectile dysfunction, or a combination of the two. Premature ejaculators showed thresholds commensurate with controls, while men with erectile dysfunction, or combined erectile dysfunction and premature ejaculation, showed significantly elevated thresholds. Although premature ejaculators did not show penile hypersensitivity, there was a significant correlation in this group between ejaculation latency and threshold. Overall, these findings argue against a primary role for penile sensitivity in ejaculation latency, and suggest that other somatic factors or cognitive factors may play the more critical role in premature ejaculation. PMID- 8246275 TI - Clomipramine versus placebo in the treatment of premature ejaculation: a pilot study. AB - This study evaluated clomipramine as a possible treatment for premature ejaculation. Twenty patients with premature ejaculation were randomly allocated to treatment with clomipramine or placebo in a double-blind study. Average estimated time to ejaculation after vaginal penetration increased to 6.1 minutes on 25 mg. of clomipramine and to 8.4 minutes on 50 mg. of clomipramine. These estimated times were significantly different from estimated time to ejaculation while on placebo. These findings suggest that low dose clomipramine may be useful in the treatment of premature ejaculation. PMID- 8246276 TI - Psychosexual functioning in women with complete testicular feminization: is androgen replacement therapy preferable to estrogen? AB - Effects of oral testosterone undecanoate (Andriol) on blood hormone levels, moods, sociosexual functioning and self-image of the body were studied in four gonadectomized patients with complete testicular feminization. In a double-blind cross-over experiment, patients were treated with oral testosterone undecanoate (120 mg/day) or placebo for four weeks. Peripheral blood was taken for hormone assays at the end of each four-week period. Psychosexual functioning was reported once weekly, mood scales twice weekly. In three patients testosterone treatment resulted in adult male blood levels of testosterone and estradiol. One patient did not show increased steroid levels, possibly because of hyperthyroxinaemia. No systematic effects were found on coitus, masturbation, sexual thoughts, scores on desire for bodily contact, and on attention for physical appearance. In one patient there was a marked and sustained rise in the positive moods and a fall in negative moods during androgen treatment. These results do not demonstrate that androgen therapy is preferable to estrogen in gonadectomized women with complete testicular feminization with regard to psychosexual functioning. PMID- 8246277 TI - Boundary issues in teacher-student relationships. AB - Increasing concern about therapist-patient sex has led to a consideration of boundaries in all trust-based relationships, which always include elements of power and dependency. Such relationships include those between teacher and student, especially those involving research or clinical supervision. Teacher student relationships differ from those between therapist and patient because of the collegiality considered important for the student's development. Yet, both share the objective of fostering independence of the "client." Therefore, teachers need to find a balance of nurturance and separateness in their relationships with their students, so that the students can carry that modeling into their own careers. PMID- 8246278 TI - Diagnosis and treatment of sexual addiction. AB - Following a brief introduction to the concept of addiction, the definition of and diagnostic criteria for sexual addiction are presented. A theoretical framework for treatment of sexual addiction is then outlined, based on an understanding of the underlying addictive process: the compulsive dependence on external actions as a means of regulating one's internal states. Effective treatment addresses both addictive behavior and the addictive process. Addictive sexual behavior is addressed through behavioral symptom management, which includes relapse prevention and other cognitive-behavioral techniques. The addictive process is addressed by enhancing self-regulatory functions through individual psychotherapy, therapeutic group experience, and pharmacotherapy (medication treatment, when indicated). An integrated system for treatment of sexual addiction, which brings together these therapeutic methods in one theoretically coherent, clinically unified approach, is outlined. PMID- 8246279 TI - K-ras gene mutation as a pathogenetic and diagnostic marker in human cancer. PMID- 8246280 TI - Human tumor-specific immunity--has it "arrived"? PMID- 8246281 TI - Persistent high hopes held for colony stimulating factors. PMID- 8246282 TI - Senators seek reform of Orphan Drug Act. PMID- 8246283 TI - Biotech firms concerned about drug pricing issues. PMID- 8246284 TI - Does abortion increase breast cancer risk? PMID- 8246285 TI - Cigarette smoking and risk of acute leukemia: associations with morphology and cytogenetic abnormalities in bone marrow. AB - BACKGROUND: Cigarette smoking may be a risk factor for leukemia. No detailed biological mechanism has been proposed, but a causal link is made plausible by evidence of systemic effects of cigarette smoke and the presence in cigarette smoke of chemicals that have been associated with leukemia risk. PURPOSE: Our purpose was to investigate the leukemia risk associated with cigarette smoking in a multicenter case-control study of acute leukemias in adults. METHODS: Adults aged 18-79 with newly diagnosed leukemia were contacted to participate in this epidemiologic study when they entered a clinical trial to be treated under protocols sponsored by Cancer and Leukemia Group B. Smoking histories for 610 patients with acute leukemia and 618 population control subjects were obtained by telephone interviews. We examined bone marrow samples and classified patients by morphology of leukocyte precursor cells according to the French-American-British (FAB) classification system and, for 378 patients, by the presence or absence of specific clonal chromosome abnormalities. We calculated odds ratios (ORs) for risk of leukemia associated with smoking cigarettes. ORs were adjusted for age, race, and sex. RESULTS: Smoking was associated with only a modest increase in risk for leukemia overall (adjusted OR = 1.13; 95% confidence interval [CI] = 0.89-1.44). However, among participants aged 60 and older, smoking was associated with a twofold increase in risk for acute myeloid leukemia (AML) (OR = 1.96; 95% CI = 1.17-3.28) and a threefold increase in risk for acute lymphocytic leukemia (ALL) (OR = 3.40; 95% CI = 0.97-11.9). Among older persons, risks increased with amount and duration of smoking. Smoking was associated with increased risk for AML classified as FAB type M2 at all ages, with ORs of 1.70 (95% CI = 1.00-2.90) for those younger than 60 and 3.50 (95% CI = 1.53-8.03) for those aged 60 and older. Smoking was also associated with ALL type L2 at all ages, with ORs of 1.72 (95% CI = 0.90-3.27) for those younger than 60 and 5.34 (95% CI = 1.03-27.6) for those who were older. Smoking was more common among patients with specific chromosome abnormalities in AML [-7 or 7q-, -Y, +13] and in ALL [t(9;22)(q34;q11)]. CONCLUSIONS: Cigarette smoking is associated with increased risk for leukemia and may lead to leukemias of specific morphologic and chromosomal types. The association varies with age. IMPLICATION: Examining discrete subtypes of disease may permit more accurate assessment of risk. As standardized morphologic classification and cytogenetic and molecular evaluation of leukemia patients becomes more common, epidemiologic studies that take advantage of these advances will begin to contribute to the identification of additional risk factors and mechanisms in acute leukemia. PMID- 8246286 TI - K-ras mutations in putative preneoplastic lesions in human colon. AB - BACKGROUND: A mutation in c-K-ras (KRAS2) has long been implicated as one of the important early events in the development of a large proportion of human colon cancers. Aberrant crypt foci, putative preneoplastic lesions identified microscopically in wholemounts of colons, have been shown to occur with high frequency in the colons of animals treated with colon carcinogens and in the grossly normal mucosas of patients with colon cancer. PURPOSE: In this study, we asked whether the mutational activation of K-ras occurs in the aberrant crypt foci of human colon. METHODS: Grossly normal colonic mucosas were obtained from seven patients during surgery and were provided to us by the Western Division of the Cooperative Human Tissue Network located at Case Western Reserve University. A total of 42 samples, consisting of aberrant crypt foci and similarly sized normal crypt areas, were microdissected from the grossly normal colonic mucosas. The DNA region containing codon 12 of K-ras was amplified by polymerase chain reaction and analyzed for mutations by dot-blot hybridization with specific oligonucleotide probes complementary to normal or mutant sequences. RESULTS: Mutations in codon 12 of K-ras were found in 11 (73%) of 15 aberrant crypt foci but not in any of 27 morphologically normal crypt areas from the same patients. CONCLUSIONS: The observed high frequency of K-ras mutations in these microscopically identifiable lesions makes mutation in K-ras the earliest identified gene-mutational event in human colon tumorigenesis, establishes that it often occurs prior to the development of polyps, and is consistent with the hypothesis that aberrant crypt foci are the earliest identified precursors of human colon cancer. IMPLICATIONS: Further analysis of aberrant crypt foci may identify yet unknown early genetic events that precede human colon cancer. PMID- 8246287 TI - Antibodies against autologous tumor cell proteins in patients with small-cell lung cancer: association with improved survival. AB - BACKGROUND: The frequency and clinical relevance of human antitumor immune responses is not well known, and few target antigens have been identified. PURPOSE: This study was designed to determine the frequency of antibodies reactive against extracts of autologous tumor cell lines and to correlate these data with survival. METHODS: Serum samples were obtained from 40 lung cancer patients treated on National Cancer Institute protocols. These sera were used as probes in immunoblots against protein extracts from tumor cell lines derived from each of these patients. RESULTS: We detected serum antibodies against autologous tumor cell proteins in 21 (58%) of the 36 patients with small-cell lung cancer (SCLC) and three (75%) of the four with non-small-cell lung cancer (NSCLC). Two patients' sera detected the p53 tumor suppressor gene product and two detected the product of the HuD gene (associated with paraneoplastic neurological syndromes) in their autologous tumor cell lysates. SCLC patients with antibodies against autologous tumor cell proteins had improved survivals compared with those in the antibody-negative group (P = .059). All patients who lived longer than 36 weeks were antitumor antibody positive. Sera from six (86%) of seven patients with limited disease were positive for antibodies that reacted against autologous tumor cells, compared with 15 of 29 (52%) of sera from patients with extensive disease. CONCLUSIONS: Our results suggest that the sera from patients with SCLC frequently contain antibodies against tumor cell proteins and that these antibodies are associated with improved survival. IMPLICATIONS: These data suggest that an antitumor immune response may affect tumor growth, and that the anonymous proteins detected by antitumor antibodies in lung cancer patient sera may represent proteins involved in the development of lung cancer or in its clinical manifestations. PMID- 8246288 TI - Mutations of the p53 gene as a predictor of poor prognosis in patients with non small-cell lung cancer. AB - BACKGROUND: Inactivation of the p53 tumor suppressor gene (also known as TP53) through a point mutation and/or loss of heterozygosity is one of the most common genetic changes found in various types of human tumors. PURPOSE: Our purpose was to investigate the relationship between the presence of p53 gene mutations and survival of patients with non-small-cell lung cancer (NSCLC) of all stages who underwent surgery with preoperative curative intent as a routine therapeutic intervention. The prognostic significance of factors like sex, age, tumor histology, and the stage of the disease was also evaluated. METHODS: We analyzed 120 tumor specimens from patients with histologically confirmed NSCLC for p53 mutations occurring in exons 5 through 8 by polymerase chain reaction-single strand conformation polymorphism assay of genomic DNA. Univariate and multivariate analyses were performed to assess the association between p53 mutations and the survival of the NSCLC patients. RESULTS: Fifty-one (43%) of 120 tumor specimens showed p53 mutations. Overall, the p53 mutations did not correlate with sex, age, or the clinical stage of the disease but showed frequent association with tumors of squamous cell histology. Univariate analysis revealed that the patients with p53 mutations survived for a significantly shorter period of time after surgery than those without the mutations (P = .0100, logrank test). The presence of p53 mutations was a significant prognostic factor in the patients with advanced disease (stages IIIA through IV) (P = .0091) but not in those with early disease (stages I and II) (P = .2837). Multivariate analysis using the Cox proportional hazards model found independent prognostic significance for p53 mutations (hazards ratio [HR] = 1.84; P = .018) and advanced disease stage (HR = 2.20; P = .003). The model also predicted the lower risk for female patients (HR = 0.51; P = .040). CONCLUSION: The occurrence of p53 mutations in some NSCLC tumors may be independently associated with a shortened overall survival and may be of somewhat more prognostic significance in patients with advanced stage than in those with early stage of the disease. IMPLICATION: Detection of p53 mutations may help in the selection of NSCLC patients suitable for appropriate investigational therapeutic strategies in view of improving their survival and quality of life. PMID- 8246289 TI - Esophageal epithelial hyperproliferation following long-term alcohol consumption in rats: effects of age and salivary gland function. PMID- 8246290 TI - The artificial restoration of a dental root. AB - Periodontal ligament (PDL) cells, osteoblasts (OB), and gingival (GIN) cells originating from human periodontium were co-cultured indirectly with human peripheral blood lymphocytes (PBL). The formation of osteoclasts (OC) from each the co-cultured PBL was compared with a standard PBL culture. A marked suppression of OC formation was observed in PBL co-cultured with PDL cells, and an enhanced OC formation was observed in PBL co-cultured with OB and GIN cells, when compared with the standard PBL culture. The suppressing activity of PDL cells and the enhancing activity of OB and GIN cells on the formation of OC derived from PBL were also found, when the co-culture fluids of PDL/PBL, OB/PBL, and GIN/PBL were added to PBL, and the numbers of OC were counted after 7 days' incubation. Furthermore, the alkaline phosphatase (ALPase) activity of PDL cells was stimulated by co-culturing them with PBL, and the ALPase activity of OB and GIN cells was inhibited by co-culturing them with PBL. When PDL cells were seeded on the surfaces of titanium discs and incubated at 37 degrees C in a 5% CO2 incubator, PDL cells could adhere faster onto titanium surfaces that were coated with a cell-and-tissue-adhesive substance than onto non-coated titanium surfaces. These cultures formed a confluent monolayer on the surfaces of titanium discs by means of an autologous serum containing alpha MEM. These results clearly suggest that the periodontal ligament is a specifically differentiated tissue whose function is to protect alveolar bone from bone resorption due to biting force.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246291 TI - Periotest to monitor osseointegration and to check the occlusion in oral implantology. AB - This paper reports on Periotest, its technique's history, and its possible applications. Periotest is a new diagnostic method, developed by an interdisciplinary research group embracing dentists, physicists, mathematicians, and computer engineers. Periotest will--apart from periodontal questions--mainly assist oral implantology. PMID- 8246292 TI - The implant-supported overdenture prosthesis for the edentulous maxilla. AB - Restoration of the edentulous maxilla with endosseous implants and a fixed prosthesis can be difficult. Problems encountered may include inadequate available bone, deficient lip support, and poor esthetics and phonetics. A palateless maxillary implant-supported overdenture has been shown to be useful in solving these problems. Four endosseous implants were placed in the anterior maxilla of a 59-year-old patient and allowed to osseointegrate for six months. After second-stage surgery was completed, a mesostructure bar incorporating ERA attachments was fabricated along with a palateless overdenture. This type of prosthesis offered a solution to the moderate to severely atrophic maxilla for a patient who was unable to tolerate a conventional prosthesis. PMID- 8246293 TI - Correction of denture fracture secondary to overextended transosteal pins in a staple implant. AB - The mandibular staple bone plate implant has been successfully used to rehabilitate the severely atrophic edentulous mandible. Numerous complications have been associated with the staple implant. The problem of overextended transosteal pins leading to repeated fracture of the mandibular denture is examined. The case report describes the technique involved in shortening the transosteal pins so that proper vertical dimension can be verified. This allows for adequate thickness of acrylic in the anterior region of the prosthesis. Proper form, function, and esthetics are established. PMID- 8246294 TI - Subperiosteal implants. AB - This paper analyzes 115 cases of subperiosteal implants placed during a 15-year period by means of the same surgical technique. The majority of implants were in women (86%) in both the maxilla and mandible. The varieties of implants placed were grouped by totals for statistical purposes: Kennedy classes I and II, Universal, and interdental. Although details of the surgical and prosthetic techniques are not included in this report, the procedures carried out by other authors are described. The majority of the implants evaluated had been in place for from 5 to 10 years (58.3%). The implants that experienced the highest levels of success are the completed designs in the mandible and the interdental in the maxilla. In both cases, the best results were observed when the antagonist was a complete-denture prosthesis. PMID- 8246295 TI - A rare find in Anatolia--a tooth implant (mid-sixth century B.C.). PMID- 8246296 TI - Pigeon-holes for dental implant providers. PMID- 8246297 TI - Porous ceramic vehicles for rat-marrow-derived (Rattus norvegicus) osteogenic cell delivery: effects of pre-treatment with fibronectin or laminin. AB - Cultured marrow-derived rat (Rattus norvegicus) mesenchymal cells can differentiate into osteoblasts when combined with an appropriate delivery vehicle and implanted in vivo. Porous ceramics have been successfully utilized for this purpose; however, the cellular interactions with these ceramics have not been delineated, and the optimal conditions for cell loading have not been determined. We have investigated the effects of ceramic pre-treatment with fibronectin and laminin on culture-expanded marrow mesenchymal cells. Scanning electron microscopic imaging was used for assessment of the interaction of these osteogenic cells with treated and untreated ceramic surfaces. The capability of pre-treated and untreated ceramics to retain an inoculum of these cells was determined by quantitation of [3H]thymidine-labeled cells added to ceramics and implanted subcutaneously into syngeneic rat hosts. Finally, the rate of bone formation in ceramic-cell composites, at harvest times ranging from 1 to 5+ wk, was determined histologically. Scanning electron microscopic images indicate that ceramics pre-treated with either laminin or fibronectin provide a surface on which marrow mesenchymal cells spread. Fibronectin and laminin increase cell retention within coated ceramics compared with uncoated controls. Furthermore, morphological evidence of osteogenesis is observed earlier in coated than in untreated ceramics. These observations indicate that cell attachment proteins, such as fibronectin, may augment bone formation in cell-ceramic composites by promoting the attachment and retention of osteoprogenitor cells within ceramic pores. PMID- 8246298 TI - Response of bone and cartilage cells to biomaterials in vivo and in vitro. AB - In vivo and in vitro models have been developed to study the bone/material interface. The in vivo model exploits the osteogenesis that accompanies marrow ablation of the rat tibia and uses morphological and biochemical changes in extracellular organelles, called matrix vesicles, as markers of the healing process. Matrix vesicles, which are associated with primary bone formation and calcification, are produced by osteoblasts and are sensitive to cellular and environmental regulation. In bone adjacent to bone-bonding implants, matrix vesicle number increases, as does its alkaline phosphatase activity. In bone adjacent to nonbonding materials, matrix vesicle activity is inhibited. The materials exert systemic effects which can also be studied by use of matrix vesicles. Cell models are needed in order for the specificity of the cellular response to the material to be understood. By the use of culture plates sputter coated with implant materials, the response of cells can be studied under controlled conditions. Comparison of the response of costochondral chondrocytes at two stages of endochondral development demonstrates that the effects of various materials are surface- and cell-maturation-dependent. Cells cultured on Ti exhibited increased alkaline-phosphatase-specific activity, whereas those cultured on Al2O3 have decreased enzyme activity. PMID- 8246299 TI - Structure of the bone interface to dental implants in vivo. AB - Dental implant materials can develop different kinds of interfaces when used for bone or tooth substitution. This paper describes the tissues and the tissue reactions of the interface created in the implantation bed between bone tissue and implants made of dense sintered hydroxylapatite. Healing was clinically and histologically evaluated during a six-month to five-year period. The light microscopic results show extensive bone apposition on the osseous sides of loaded implant cylinders and also on subperiosteally placed granules of dense hydroxylapatite. The presence of the loaded implant did not disturb the normal bone physiology. Bone turnover also included the bone tissue toward the implant surface. At the ultrastructural level, an amorphous collagen-free zone of about 200 nm was frequently observed which was separated from the surrounding mineralized extracellular matrix by a thin, electron-dense layer of about 20-100 A. Contact microradiographs showed an intimate bond to bone. The bone growth extending up the implant cylinder was always accompanied with an embedding of collagen fibers and the presence of a well-developed and complete new gingival attachment apparatus. This was confirmed by scanning electron microscopy. However, any inflammatory reactions in the transmucosal area were accompanied with concomitant bone resorption and loss of bone support. PMID- 8246300 TI - Age factor in human alveolar bone repair. AB - This was a retrospective review study based on histological and histochemical observations of 69 biopsy specimens of human extraction wounds, screened for elimination of medical or nutritional disorders that might have interfered with normal healing. Initially, for the first 10 post-extraction days, tissues from the younger individuals (second decade or less) and those from older individuals (sixth decade or over) appeared to heal at the same rate. After 10 post extraction days, the younger tissues continued with progressive healing and osteoid formation, while the older tissues remained in a resting (lag) phase. Following approximately three weeks, the older tissues entered into an accelerated growth cycle, and by 30 to 40 days equaled the healing pattern of the younger tissues, describing a sigma-type curve. PMID- 8246301 TI - Immediate implantation into the posterior maxilla after antroplasty: the Cranin Russell Operation. AB - Heretofore, the absence of adequate alveolar bone (4 mm or less) beneath the maxillary sinus has prevented placement of endosteal root-form implants at the same time as antroplasty. A technique has been devised which permits implantation to occur upon the occasion of sinus floor elevation. This has been made possible by the use of titanium fracture miniplates, which serve as transitional fixation devices until mineralization of the graft material creates implant osseointegration. PMID- 8246302 TI - Dental implant in resorbed root. PMID- 8246303 TI - Know when to say "when". PMID- 8246304 TI - Experimental studies of the implant-tissue interface. AB - This comparative study analyzed the epithelial, gingival connective tissue, and osseous tissue interface with clinically and radiographically integrated endosteal dental implants. Undecalcified interfacial tissues were sectioned for both routine transmission electron microscopy (TEM) and for High Voltage Electron Microscopy (HVEM). A protective perimucosal biological seal was formed by regenerating soft tissues (epithelium and connective tissue). Inferior to this protective soft-tissue attachment seal, the apical support complex was shown to vary in morphology. Mineralized bone was closely apposed to significant regions of the implants, separated only by an electron-dense deposit of approximately 20 nm. Osteoblasts were observed adjacent to the implant, as were osteocytes within the underlying supporting bone. Osteoblasts were observed associated with a connective tissue stroma adjacent to the existent mineralized bone. Osteocyte cellular processes extended toward adjacent osteocytes, toward vascular elements, and directly to the implant surface. These observations demonstrate the healthy interface of mineralized tissues with both root-form and blade implants. Mineralization patterns of the bone supporting the implants appeared consistent with normal mandibular maturation patterns. PMID- 8246305 TI - The developmental stages of osteoblast growth and differentiation exhibit selective responses of genes to growth factors (TGF beta 1) and hormones (vitamin D and glucocorticoids). AB - Osteoblasts in vitro undergo a developmental sequence of growth and differentiation characterized by a stage-specific expression of cell growth and bone-related genes. Our studies show that contributions to development of the osteoblast phenotype include: (1) the down-regulation of proliferation, (2) biosynthesis and organization of type 1 collagen extracellular matrix, and (3) mineralization of this matrix. Hormones and growth factors added to isolated osteoblasts during the initial proliferation period can dramatically alter the subsequent program of developmental stages of differentiation. Studies are presented that demonstrate alterations in gene expression in response to: (1) transforming growth factor beta (TGF beta 1), that prevents differentiation of the cells and their ability to form bony tissue-like nodules; (2) glucocorticoids, which induce and accelerate development of the bone-cell phenotype; and (3) the active metabolite of vitamin D, 1,25(OH)2D3, which results in both inhibitory and stimulatory effects of the hormone on growth and differentiation--effects that are dependent upon the differentiated state of the osteoblast. Thus, in the presence of these physiological mediators of bone-cell phenotype development, many of the growth and differentiation relationships operative in osteoblasts are modified or abrogated, precluding expression of the structural and functional properties of mature bone cells and tissue. These findings have significant implications for use of hormones or growth factors in the treatment of disorders where bone is involved. PMID- 8246306 TI - A new visual method for evaluating multiple data (Diagnostic Radar Chart) in general toxicological study. AB - A Diagnostic Radar Chart method was devised for use in the general assessment of the toxicity of drugs on the basis of visual patterns in toxicity studies. Its usefulness was evaluated on the findings of 32 laboratory items in 3 groups of animal models with experimental pathological conditions. The chart was found to facilitate intuitive discrimination among three types of pathological animal models, evaluation of sexual differences and evaluation of individual differences among the animals for each item. Therefore, if patterns are constructed on the Diagnostic Radar Chart for each of the drugs usable for preparing animal models with typical experimental pathological conditions, the chart is thought to be useful for evaluation or identification of the toxicity of an unknown drug. PMID- 8246307 TI - The effect of food chemicals on cell aging of human diploid cells in in vitro culture. AB - The potency of food chemicals to induce cell aging was evaluated in human diploid fibroblast cells HAIN-55 having a finite replicative potential by using in vitro aging markers, i.e., decreases of maximum proliferative potential (lifespan) of cells, saturation density in monolayer culture (SD), plating efficiency (PE) and mitotic index (MI), and an increase of cells with polyploid karyotypes. By treatment twice with low concentration of genotoxic chemicals aflatoxin B1, allylisothiocyanate or trans-cinnamaldehyde (severe clastogenic flavoring agent; Kasamaki et al., 1982), lifespan (expressed by the number of cumulative cell population doubling (CPD)) of the treated cells was reduced by 8-12 CPDs accompanied by change of the other aging markers. By successive treatment (29 or 25 times) with non-genotoxic chemical aspartame (N-L-aspartyl-L-phenylalanine) or L-canavanine (structural analogue of L-arginine), lifespan of the treated cells was also slightly shortened (by 2-6 CPDs) compared with the untreated control cells. In the process of cell aging, Mitochondrial activity (MTT activity) decreased almost in parallel with the decrease of SD and MI. On the basis of these results, a variety of genotoxic and non-genotoxic chemicals were examined by using MTT activity as the aging marker for their effects on the aging of HAIN 55 cells and bovine artery endothelial cells which also had a finite replicative potential. The results showed that seven genotoxic and nine non-genotoxic chemicals promoted cell aging. PMID- 8246308 TI - Histometric changes in connective fibers of the lung following paraquat administration in rats. AB - Pulmonary fibrosis induction by paraquat was studied histometrically using an image analyzer. Rats were given a subcutaneous injection of paraquat (7 or 25 mg/kg) and autopsied 3, 6 or 9 days later. The percentage of the area of connective fibers per unit area, and lung prolyl hydroxylase activity, as a biochemical parameter, were measured. Rats treated with 25 mg/kg of paraquat showed an increase in the percentage of the area of the connective fibers per unit area and also an increase in lung prolyl hydroxylase activity 6 days later. Thus, pulmonary fibrosis was confirmed both morphologically and biochemically. It is concluded that histometric examination using an image analyzer provides a very accurate method for the evaluation of pulmonary fibrosis. PMID- 8246309 TI - A new potent teratogen in CD rats inducing cleft palate. AB - The teratogenic potential of cis-1-[4-(p-menthane-8-yloxy)phenyl] [corrected] piperidine (YM9429) was evaluated using CD (SD) rats. YM9429 induced cleft palate and specific skeletal variations including accessory cervical and lumbar ribs or excessive formation of the 7th lumbar vertebra by oral treatment during the organogenetic periods. No visceral or external malformations were induced, and no embryo/fetal mortality or fetal growth retardation was observed. Maternal plasma biochemical examination revealed decreases of cholesterol and phospholipid levels during days 15-17 of pregnancy after the treatment. The results suggest that YM9429 is a potent and specific teratogen inducing cleft palate in CD rats, and the reduced maternal plasma levels of cholesterol and phospholipid during the period of palatine closure are related to the induction of cleft palate. PMID- 8246310 TI - Studies on the therapeutic effect of 2-pyridine aldoxime methiodide (2-PAM) in mammals following organophosphorus compound (OP)-poisoning (report II): aging of OP-inhibited mammalian cholinesterase. AB - We studied the ability of 2-PAM to reactivate cholinesterase (ChE) inhibited by organophosphorus compounds (OPs) and aging. We estimated the reactivation rate with 2-PAM following inhibition of ChE by fenitrothion, methylparathion or ethylparathion using erythrocytes of rat and rabbit and rat brain. The period of time during which inhibited ChE could be reactivated was shorter in the case of inhibition by fenitrothion or methylparathion than in the case of inhibition by ethylparathion. This results suggest that aging is related to the presence of the alkyl group in OPs, and occurs faster in the case of inhibition by OPs with an O,O-dimethyl moiety than in the case of inhibition by OPs with an O,O-diethyl moiety. PMID- 8246311 TI - Preventive effects of diphenyl dimethyl dicarboxylate on the immunotoxicity of carbon tetrachloride in ICR mice. AB - To investigate the preventive effects of diphenyl dimethyl dicarboxylate (PMC) on the immunotoxicity of carbon tetrachloride (CCl4) in ICR mice, PMC (3 and 6 mg/kg, respectively) were orally administered to mice through a zonde once a day for 28 consecutive days. CCl4 solution was also administered at 1 ml/kg (25%) p.o. 2 hr later the administration of PMC twice a week. Mice were immunized and challenged with sheep red blood cells (SRBC). Immune function were evaluated by humoral, cell-mediated and non-specific immune responses. The results of this study were summarized as follows; 1. The body weight gains and the relative spleen and thymus weights were significantly increased by PMC treatment as compared with treatment of CCl4 alone. However, the relative liver weights were significantly decreased. These values were similar to those of control mice. 2. Hemagglutination (HA) titers were significantly enhanced by PMC treatment as compared with treatment of CCl4 alone. Plaque forming cells (PFC) were also greatly increased by PMC treatment, especially at a dose of 6 mg/kg of it. 3. Delayed-type hypersensitivity (DTH) response was significantly decreased in mice treated with CCl4 and PMC along with the increase of PMC doses, as compared with those in the mice treated with CCl4 alone, while rosette forming cells (RFC) were significantly increased. These results were similar to those of control mice. 4. In PMC treatment as compared with treatment of CCl4 alone, there were significant increases in activities of natural killer (NK) cells and phagocytes along with circulating leukocytes. These results suggest that PMC has significant preventive effect on CCl4-induced immunotoxic status. PMID- 8246312 TI - Acute toxicity of cadmium in rats with or without cadmium pretreatment. AB - The acute toxicity of cadmium (Cd) in male rats was examined with or without Cd pretreatment. Firstly, the metallothionein (MT) contents in the liver and kidney after Cd exposure (2.0 mg/kg, i.v.) were determined. The MT contents in the liver increased immediately to a peak (36.0 +/- 5.5 n mol/g wet tissue) 2 days after Cd exposure and were 55-fold higher than that at 0 day (0.64 +/- 0.25 n mol/g wet tissue). On the other hand, the MT contents in the kidney increased slightly but steadily for 14 days after Cd exposure. In the study for comparison of Cd-induced toxicity, the LD50 value of the Cd-pretreatment group (Group II) was approximately 2-fold higher than that of the non-pretreatment group (Group I). In microscopic findings, differences between rats in Group I and Group II were recognized in the kidney. Cytoplasmic vacuolation of the proximal tubular epithelium in the kidney was observed in Group I, while degeneration or coagulative necrosis in the proximal tubular epithelium was observed in some rats in Group II in addition to the cytoplasmic vacuolation. Because the toxic changes other than in the kidney in Group II were almost equal to or less than that in Group I, in spite of the doubled dosage of Cd, the toxic effects of Cd, except on the kidneys, were considered to be reduced by the pretreatment with Cd. PMID- 8246313 TI - Effects of rokitamycin on bilirubin-albumin binding: a study in vitro. AB - The effects of rokitamycin, a macrolide antibiotic, on the binding of bilirubin to albumin were examined in vitro using blood plasma from young rats with hyperbilirubinemia, bilirubin-supplemented serum separated from human cord blood, and the human serum from a newborn infant with icterus gravis neonatorum. Rokitamycin at concentrations from 1 to 500 micrograms/ml (the entire range over which experiments could be conducted) had little or no effect on the concentration of unbound bilirubin in any of the incubation mixtures used. This result suggests that rokitamycin has no effect on the binding of bilirubin to albumin. PMID- 8246314 TI - High susceptibility to aflatoxin B1 and benzo[a]pyrene of BALB3T3 A31-1-1 cells expressing monkey CYP1A1. AB - The monkey CYP1A1 has been expressed in BALB 3T3 A31-1-1 cells and the expressed proteins were assayed for their capacity to activate aflatoxin B1 (AFB1) and benzo[a]pyrene (B[a]P). The transformed cells were approximately 5.4- and 4.7 fold more sensitive to AFB1 and B[a]P than the parental cells, respectively. These results indicate that monkey CYP1A1 cDNA encoded a functional protein and that the expressed CYP1A1 enzyme is active for the activation of B[a]P as well as AFB1 to produce toxic metabolites. PMID- 8246315 TI - Substance abuse in adolescent mothers: strategies for diagnosis, treatment and prevention. AB - Adolescent drug addiction and alcohol abuse are on the rise in this country. Teenagers of all races, socioeconomic groups and both genders have become avid experimenters with a full spectrum of substances, including alcohol, marijuana, stimulants, cocaine and hallucinogens. A diagnosis of addiction requires familiarity with the normative trends of adolescent behavior, as well as deviations from these trends. Moreover, an adolescent mother with a drug addiction problem confronts the additional burden of fostering adaptive developmental patterns in her infant. Treatment techniques such as previewing, which empowers through the representation and enactment of future outcomes, have helped addicted teenage mothers relinquish drug use and adopt more mature caregiving behaviors. PMID- 8246316 TI - The antecedents of women's crack cocaine abuse: family substance abuse, sexual abuse, depression and illicit drug use. AB - Young African-American women are the fastest growing group of crack cocaine users in the United States. Despite this increase, relatively little is known about women who use crack cocaine and the possible reasons for their drug use. This descriptive, exploratory study focused on data from a sample of 105, predominantly African-American women who use(d) crack cocaine, of whom 60 were in drug treatment and 45 were still using crack cocaine. Significant correlations were found between variables such as family drug use, first age of sexual abuse, age of first depressive symptoms and age of first illicit drug use. These data are consistent with other studies which have noted high rates of depression and sexual abuse in samples of predominantly Euro-American, alcoholic women. Implications for the treatment of women are discussed. PMID- 8246317 TI - Enhancing addiction treatment through psychoeducational groups. AB - Clients with an addiction disorder generally present with an ego deficit in the area of self-care. This deficiency manifests itself in an inability to suffer and struggle with day-to-day problem solving. Outpatient addiction treatment today places significant resources in psychotherapy and psychodynamic group therapy often at the expense of teaching basic life skills our clients need to negotiate day-to-day living. To address both issues of ego and life skills deficits, the use of a psychoeducational group is presented. This specialized, task-oriented didactic group experience is a necessary component of a comprehensive addiction treatment program. A psychoeducational group in the treatment of addictions can serve as a synthesis for problem-solving skills training used in mental health and the psychodynamic theory of addictive behavior. Cases are presented to illustrate the efficacy of psychoeducational groups. PMID- 8246318 TI - Psychiatric histories of drug using mothers: treatment implications. AB - One hundred forty six crack/cocaine using mothers in New York City were interviewed in a cross sectional study about life histories and drug related behaviors. Forty one (28%) reported histories of previous psychiatric medication or hospitalization. These women were significantly more likely than the rest of the sample to currently be in drug treatment; to have sexual abuse histories; and to be currently involved with men who urged them to use crack during pregnancy. Within this group, two subgroups were distinguishable: one, who had been sexually abused and initiated drug use early, and the other whose psychiatric and drug use histories were not associated with sexual abuse. The implications of these findings for screening, treatment planning, and future research are discussed. PMID- 8246319 TI - Hazards associated with the use of benzodiazepines in alcohol detoxification. AB - While benzodiazepines possess many positive attributes for the treatment of alcohol withdrawal, they are not ideal drugs. The occurrence of toxic effects is not infrequent, and unless recognized promptly, toxicity may significantly impact treatment and length of stay. Use of shorter acting benzodiazepines may result in seizure activity under certain conditions of withdrawal. PMID- 8246320 TI - Use of 'very low-dose phenobarbital' to investigate compliance in patients on reducing doses of methadone (detoxification). AB - Incorporation of very low doses of phenobarbital into a methadone linctus has enabled us to monitor the compliance of 7 patients receiving a reducing dose of methadone (detoxification) for treatment for opioid addiction. By measuring both plasma phenobarbital and methadone we detected 4 patients who consumed extra illicitly obtained methadone during the detoxification regime. Treatment outcome was poor; 11 of the original 18 patients dropped out of treatment within 14 days and of those who remained, 4 patients relapsed and reabused illicit drugs and 2 returned to a fixed dose of methadone. Laboratory measurements were successfully used to detect poor methadone compliance. PMID- 8246321 TI - A substance-abuse research-treatment clinic: effective procedures and systems. AB - Drug abuse treatment clinics vary widely. This article presents a model of a research-treatment clinic in which effective procedures and systems were instituted. These systems produce an environment in which both excellent research can be conducted and effective treatment provided, by a team of researchers and clinicians. Many of the procedures overcome frequently occurring problems at drug treatment clinics. The potential applicability of these procedures and systems to standard drug treatment clinics is discussed. PMID- 8246323 TI - Computed tomography assessment of ground-glass opacity: semiology and significance. AB - Among the computed tomography (CT) signs of parenchymal lung disease, the ground glass pattern is the one most difficult to diagnose and most influenced by CT technique. Ground-glass opacity may result from changes in the airspaces or interstitial tissues in acute or chronic infiltrative lung disease. It may also be seen as a consequence of increased capillary blood volume in redistribution of blood flow due to airway disease, emphysema, or pulmonary thromboembolism. Definition of this sign on high-resolution CT (HRCT) images, its various HRCT patterns, and potential pitfalls in its recognition are described with special attention to optimal HRCT technique. PMID- 8246322 TI - The Beech Hill Hospital Eating Disorders Treatment Program for Drug Dependent Females: program description and case analysis. AB - The association of eating disorders and substance abuse has increasingly been noted among chemically dependent women. Without diagnosis and focused interventions around the disordered eating, female substance abusers are vulnerable to chemical relapse and continued out of control eating. This paper describes an eating disorder treatment program for drug-dependent females during the inpatient phase of chemical dependency treatment. The interrelationship and need to treat both addiction and eating disorder at the beginning of early sobriety is emphasized. PMID- 8246324 TI - High-resolution computed tomography--physiologic correlation. AB - High-resolution computed tomography (HRCT) as a tool for investigation of bronchovascular and pulmonary responses to various physiologic and pharmacologic stimuli is a new field of application. The potential of this method has only recently been investigated in animal experiments. To date, research has focused on the determination of airway responses in the context of agonist challenge such as aerosolized or i.v. histamine, isotonic saline, halothane anesthesia, and hypoxia. Likewise, physiologic HRCT has been used in the elucidation of the pulmonary circulatory response to acute hypervolemia and hypoxia. Early results indicate that significant observations can be derived from HRCT as it is the only existing method that not only detects physiologic responses but, unlike existing methods, can characterize their site and locoregional differences. In this article, the rationale for and present status of physiologic HRCT is presented. PMID- 8246325 TI - Dynamic imaging of lung morphology with ultrafast high-resolution computed tomography. AB - To aid in the diagnosis of lung diseases associated with airway obstruction and air-trapping, we use dynamic ultrafast high-resolution computed tomography (DUHRCT). We obtain a rapid series of ultrafast HRCT scans during a forced inspiratory and expiratory maneuver to demonstrate dynamic lung attenuation changes. Alteration in lung morphology can be seen coincident with changes in lung attenuation during different phases of the respiratory cycle. Lung attenuation changes can be evaluated qualitatively or quantitatively by using time-attenuation curves measured for specific regions of lung. In a normal population, we noted that the lingula is a common site of unsuspected and asymptomatic air-trapping. In subjects with airway obstruction, and in those with cystic lung disease, air-trapping is readily detected, even at the secondary pulmonary lobule level. In some of those cases with air-trapping, a paradoxical decrease in lung attenuation was noted during forced exhalation. Recent development of spiral CT scanners may allow more common usage of a similar dynamic technique. Simple paired inspiratory and expiratory scans, obtained during suspended respiration at the same level, may be sufficient for the detection of air-trapping in many patients. PMID- 8246326 TI - Spiral computed tomography in the assessment of focal and diffuse lung disease. AB - Conventional computed tomography (CT) processes the raw data of a single rotation at any given level to produce the image at that level. Scan collimation and interspacing are determined prospectively. Spiral CT combines longitudinal patient transport with continuous data acquisition. Interpolated projections are used for reconstructing any level inside the scanned volume. Therefore, spiral CT allows any number of images, their spacing, position, and field of view to be determined retrospectively. Technically, the only limiting factors of spiral CT are the load of the x-ray tube and the computation power of the scanner. Spiral CT is superior to conventional CT in detecting lung nodules, in imaging the representative slice of a small nodule by retrospective selection, and in its ability to reformat the image in any plane. It also allows rapid scanning and three-dimensional re-formation of images. It may reduce the dose of contrast agent required for optimal enhancement of the vascular structures, reduce the radiation dose, and eliminate motion artifacts. PMID- 8246327 TI - Variables affecting pulmonary nodule detection with computed tomography: evaluation with three-dimensional computer simulation. AB - To meaningfully evaluate factors determining the overall accuracy of computed tomography (CT) for identifying pulmonary nodules, computer-generated nodules were superimposed on normal CT scans and interpreted independently by three experienced chest radiologists. Variables evaluated included nodule size, shape, number, density, location, edge characteristics, and relationship to adjacent vessels, as well as technical factors, including slice thickness and electronic windowing. The overall sensitivity in identifying nodules was 62% and the specificity was 80%. On average, the observers identified 56, 67, and 63% of nodules on 1.5-, 5-, and 10-mm-thick sections, respectively (p = 0.037). Nodules were more difficult to identify on 1.5-mm-thick sections. On average, observers identified 1, 48, 82, and 91% of nodules < 1.5, < 3, < 4.5, and < 7 mm in diameter, respectively (p < 0.001). Other factors that made a significant contribution (p < 0.01) in identifying nodules, as determined by linear discriminant function analysis, included nodule location, angiocentricity, and density. We concluded that computer-generated nodules can be used to assess a large number of imaging variables. We anticipate that this approach will be of considerable utility in assessing the accuracy of interpretation of a wide range of pathologic entities as well as in optimizing three-dimensional scan protocols within the thorax. PMID- 8246328 TI - High-resolution computed tomographic bronchiolography using perfluoroctylbromide (PFOB): an experimental model. AB - The use of perfluoroctylbromide (PFOB), a liquid ventilatory agent, was evaluated as a computed tomographic contrast agent to visualize small airway anatomy to the level of the centrilobular bronchiole, normally not visible on high-resolution computed tomography (HRCT). A freshly excised neonatal lamb heart-lung preparation was tracheally intubated, suspended in a saline bath, and mechanically ventilated with gas. After obtaining HRCT control images with suspended respiration using continuous positive airway pressure, 30 ml of perfluorocytlbromide was instilled into the trachea and repeat scans were obtained. These images demonstrated PFOB filling and distending the airways to the level of the centrilobular bronchioles and their first order branches. There was only minimal spillage into air spaces, allowing excellent anatomic detail of the bronchiolar structures. A liquid ventilatory agent, PFOB is a superb candidate as a bronchographic contrast agent due to its promotion of gas exchange, low toxicity, low surface tension, radiopacity, and vaporized excretion via the lung. It has great potential to evaluate small airway disease when used in conjunction with HRCT. PMID- 8246329 TI - Pulmonary calcinosis following orthotopic liver transplantation. AB - Pulmonary opacities following orthotopic liver transplantation (OLT) are frequent due to pneumonia, infarction, or pulmonary edema. Metastatic pulmonary calcifications are present as persistent opacities that may mimic these conditions. In a series of 91 patients who underwent OLT, chest radiographs of 77 were reviewed and pulmonary calcinosis was seen in 4 (5.2%). Pulmonary calcinosis may be due to a variety of conditions, including dystrophic calcification in damaged lung and primary or secondary hyperparathyroidism. In this series, patients with pulmonary calcinosis had significantly higher levels of serum phosphate and calcium postoperatively and had received more intraoperative platelets and other blood products containing exogenous calcium than other patients. Pulmonary calcinosis should be considered in patients following OLT when stable, nonspecific pulmonary opacities are present. CT or radionuclide studies will aid in confirming this diagnosis. PMID- 8246330 TI - Rounded atelectasis and fibrotic pleural disease: the pathologic continuum. AB - Although rounded atelectasis was recognized in 1928, its relationship to fibrotic pleural disease almost has gone unnoticed. We present four cases of lobar rounded atelectasis that help to clarify this relationship. Rounded atelectasis is part of the spectrum of fibrous pleural disease. Any portion of lung may become entrapped by the pleural process. When focal areas are involved rounded atelectasis results. When a larger portion of the thorax is involved with this process, lobar rounded atelectasis may occur. Very extensive disease is recognized as fibrothorax. PMID- 8246331 TI - Loculated mediastinal chylothorax resulting from esophagogastrectomy: a case report. AB - The thoracic duct is closely related to the esophagus and the pleura. During an esophagogastrectomy, the thoracic duct is at risk for injury, which usually results in chylous pleural effusions. We present an uncommon situation, however, where the pleura was not violated and a large mediastinal lymph collection resulted. PMID- 8246332 TI - Spiral volumetric CT as a routine technique for thoracic imaging. AB - The use of spiral volumetric computed tomography (CT) for thoracic imaging is described. A phantom experiment using nodules made of fish meat established a clinically acceptable practical table speed of 15 mm/s. With this table speed it was possible to scan the entire lung during a single breath-hold of 16-20 s. Comparison of this technique with conventional CT in 18 patients with 62 nodules showed no difference in effectiveness. The imaging of linear and reticular opacities, however, was not satisfactory in three of the eight lesions. Visualization of normal mediastinal vessels on spiral CT using less contrast material was comparable to that of conventional CT in 20 of 30 patients; it was less satisfactory in 10 patients, but vessels still could be identified. We conclude that spiral CT is a useful method to examine the thorax. PMID- 8246333 TI - High-frequency endoluminal ultrasonography of the esophagus in human autopsy specimens. AB - A 20 MHz ultrasound transducer housed in a 6.2 Fr catheter was used to image human esophageal autopsy specimens from six cadavers. Histologic sections taken from the areas imaged were correlated with cross-sectional sonographic images. Six echo layers were seen in the non-fluid-filled esophagus whereas seven echo layers were seen in the fluid-filled esophagus. These seven layers correspond to the following histologic structures: first hyperechoic layer--mucosa (including squamous epithelium and lamina propria); second thin hypoechoic layer--muscularis mucosae; third very bright hyperechoic layer--submucosa; fourth hypoechoic layer- circular smooth muscle; fifth thin hyperechoic layer--intermuscular connective tissue; sixth hypoechoic layer--longitudinal smooth muscle; seventh hyperechoic layer--adventitia. PMID- 8246334 TI - Quantitative assessment of sonographic image echogenicity by transmission densitometry: fetal liver model. AB - A mathematical method is proposed to compare density values of different sonographs of the same tissue. Two transformations were needed: (1) calibration of the gray scale according to the maximal range of scanning for a given tissue, and (2) comparison of the effect of two time-compensated gains on the difference in density and standardizing this effect to an arbitrary gain. To verify these relationships, 12 fetal livers were scanned at 38 weeks' gestation, at three different gains corresponding to the optimal gain and to the minimal and maximal gains that allowed a clear image to be obtained. After the mathematical transformations, the initially completely different densities were nearly identical (t-test: P = 0.9998; Wilcoxon test: P > 0.5; Pearson's R = 0.8933). This method permits an accurate quantitative assessment of the echogenicity of any parenchymatous organ. PMID- 8246335 TI - Ultrasonographic diagnosis of hepatobiliary ascariasis. AB - Eight cases of hepatobiliary invasion of Ascaris lumbricoides into the hepatobiliary tract are presented. In the presence of nonspecific symptoms and biochemical test results, the diagnosis was made in seven patients by sonography. In the remaining patient, in whom the adult roundworm was not visible on sonography owing to the presence of aerobilia, the diagnosis was made at ERCP. Sonography revealed typical tubular, echogenic, filling defects along the long axis of a dilated common bile duct, sometimes exhibiting slow movements and containing a central sonolucent line. Repeated sonographic examinations accurately predicted treatment success. PMID- 8246336 TI - Usefulness of ovarian volume and cysts in female isosexual precocious puberty. AB - The real-time pelvic sonograms of 32 girls under 8 years old with true isosexual precocity (23 cases), pseudosexual precocity (four cases), premature adrenarche (four cases), and an undetermined problem (one case) were evaluated retrospectively for ovarian volume and presence and size of cysts. Sonographic findings were compared to those of 181 age-matched controls to determine the best sonographic indicator of precocious puberty. Ovarian volume was 4.6 cm3 in girls with true isosexual precocity, 4.1 cm3 in girls with pseudosexual precocity, and less than 1 cm3 in the other patients as well as in the control population. Ovarian enlargement was bilateral in true precocity and unilateral in pseudosexual precocity. Of the 181 subjects in the control group, 96 (53%) had ovarian cysts, almost all of which were small (less than 9 mm in diameter). Ovarian cysts occurred in 22 of 32 patients (69%) with precocity. The cysts generally were smaller than 9 mm in true isosexual precocity and larger than 9 mm in pseudosexual precocity. In conclusion, small ovarian cysts are not specific to precocious puberty and its various subtypes. Bilateral ovarian enlargement appears to be a reliable indicator of true isosexual puberty, whereas unilateral ovarian enlargement in combination with macrocysts is suggestive of pseudosexual precocity. PMID- 8246337 TI - Growth retardation in prenatally diagnosed cases of gastroschisis. AB - Gastroschisis is a rare congenital anomaly characterized by the herniation of fetal intestines directly through an abdominal wall defect. It is associated infrequently with chromosomal or other nonbowel defects and can be treated surgically after delivery, with survival rates reported to be between 87 and 100%. We reviewed 21 cases of prenatally diagnosed gastroschisis to ascertain the effect of fetal growth retardation on perinatal outcome. Ten of the 21 fetuses (48%) were identified prenatally as growth retarded, although only seven of these ten truly had birth weights less than the 10th percentile. Three additional fetuses that had not been identified prenatally as growth retarded did, in fact, have birth weights less than the 10th percentile, for a total frequency of growth retardation at birth of 48% (10/21 fetuses). When compared to non-growth-retarded fetuses with gastroschisis, fetuses who were growth retarded, although more likely to have been delivered by emergency cesarean section, had shorter hospitalization times, were more likely to have undergone primary closure on the first day of life, and had fewer major complications. We conclude that growth retardation is common in fetuses with gastroschisis and the postnatal outcome in gastroschisis is not poorer for fetuses who are growth retarded. PMID- 8246338 TI - Posttraumatic "high-flow" priapism evaluation with color flow Doppler sonography. AB - Three cases of posttraumatic, high-flow arterial priapism are reported. In each, targeted color flow sonography was performed, demonstrating the site of intracavernosal arterial-sinusoidal fistula. Two of the three lesions could only be detected with the use of color Doppler sonography, emphasizing its role in the noninvasive diagnostic imaging work-up of this disorder. PMID- 8246339 TI - Differentiation of breast tumors by ultrasonic tissue characterization. AB - The ability of ultrasonic tissue characterization to differentiate and classify benign and malignant breast tissues in vivo in patients with palpable breast masses and in vitro in excised breast tissue was evaluated. One-hundred and twenty-four in vivo and 89 in vitro studies were performed using a technique of UTC based on parameters from the power spectrum of backscattered echoes. Sensitivities and specificities for diagnosing carcinoma were 86 and 84% for in vivo studies and 94 and 92% for in vitro studies. These UTC parameters provided threshold values for color-coding breast lesion images. The results of this preliminary investigation suggest that UTC provides a basis for assessing more accurately lesions suspected of being malignant prior to biopsy and possibly for evaluating breast lesions noninvasively. PMID- 8246340 TI - Color duplex sonographically guided obliteration of pseudoaneurysm. AB - Over a 21 month period, 112 patients were seen in the vascular laboratory for evaluation and therapy of possible pseudoaneurysm. Pseudoaneurysm was confirmed by color flow sonographic imaging in 31 patients. Twenty-eight of these patients underwent sonographically monitored extrinsic manual compression to induce thrombosis and subsequent obliteration. Complete manual thrombosis was achieved in 17 patients while partial thrombosis with subsequent spontaneous closure developed in three patients, for a total success rate of 71% (20/28). In eight patients attempts to thrombose the pseudoaneurysm failed (29%). Induction of thrombosis by manual compression was successful in the presence of oral and intravenous anticoagulants. No therapeutic complications were encountered. Under proper sonographic guidance, the technique of manual obliteration of pseudoaneurysms provides a safe and effective alternative to surgical intervention. PMID- 8246341 TI - Villous hyperplasia of the gallbladder: an unusual sonographic appearance. PMID- 8246342 TI - Fluctuating hemodynamics in spontaneous dissection of the cervical internal carotid artery. PMID- 8246343 TI - Sonographic appearance of craniofacial dysostosis (Crouzon syndrome) in the second trimester. PMID- 8246344 TI - Massive edema of an ovary previously fixed to the pelvic side wall. PMID- 8246345 TI - Is ST/HR slope a predictor of coronary artery lesion in effort angina pectoris? A comparison of exercise hemodynamic variables. AB - This study evaluated the clinical value of the ST/HR slope and exercise hemodynamic variables for the severity of coronary artery lesions. Twenty-nine patients with effort angina pectoris underwent ergometer exercise during right side heart catheterization. The ST/HR slope failed to differentiate three groups stratified by the number of involved coronary vessels. The ST/HR slope was uncorrelated with the Gensini score (r = 0.24). By contrast, pulmonary capillary pressure (PCP), cardiac output index (CI) and CI/PCP at peak exercise correlated significantly with the Gensini score (r = 0.73, 0.53 and 0.69, respectively). We conclude that the ST/HR slope is not a useful predictor of the severity of coronary artery lesions, while exercise hemodynamic variables measured by right side heart catheter are useful. PMID- 8246346 TI - Intracellular magnesium deficiency in acute myocardial infarction. AB - It has been hypothesized that intracellular magnesium deficiency is a pathogenetic factor in acute myocardial infarction. This study examined the time course of changes in the erythrocyte magnesium concentration and the correlation between the erythrocyte magnesium concentration and the severity of acute myocardial infarction in 49 consecutive patients with transmural acute myocardial infarction. The data were compared with results from 20 control patients without ischemic heart disease. The erythrocyte magnesium concentration (mg/dl) decreased significantly during the acute phase of the infarction (4.86 +/- 0.09 on day 1, 4.89 +/- 0.10 on day 2 and 4.86 +/- 0.10 on day 3 versus 5.26 +/- 0.19 for controls, all P < 0.05) and then normalized gradually to 5.25 +/- 0.10 on day 28. The serum magnesium concentration (mg/dl) also decreased significantly during the acute phase of the infarction (1.93 +/- 0.04 on day 1 and 2.11 +/- 0.03 on day 2 versus 2.26 +/- 0.08 for controls, all P < 0.05), before recovering to 2.28 +/- 0.06 on day 28. There were significant correlations between the erythrocyte magnesium concentration on day 1 and maximal values of serum cardiac enzymes (r = -0.30 for creatine kinase, r = -0.34 for glutamic oxaloacetic transaminase and r = -0.57 for lactate dehydrogenase, all P < 0.05). Moreover, the erythrocyte magnesium concentration was significantly lower in patients with (4.32 +/- 0.08 mg/dl, n = 13) than in those without (5.06 +/- 0.09 mg/dl, n = 36, P < 0.0001) serious arrhythmias. These data indicate that intracellular magnesium deficiency is involved in the acute phase of myocardial infarction. PMID- 8246347 TI - Theoretical study related to left ventricular energetics. AB - This study presents new mathematical relations to link oxygen consumption to different areas under the end-systolic pressure-volume relation (ESPVR). The approach consists of approximating the relation between oxygen consumption and left ventricular pressure by a quadratic polynomial, and then relating the coefficients of the quadratic polynomial to different areas under the ESPVR. The procedure applies to both ejecting contraction and isovolumic contraction. The concept of external energy reserve, EER, is introduced and experimental data that corroborate the derived theoretical results are discussed. The clinical significance of the results obtained enhances the potential possibility of using ESPVR in clinical work. PMID- 8246348 TI - Experimental creation of complete A-H block. AB - A new technique to block conduction from both atria to the atrio-ventricular node (A-V node) was devised through anatomical considerations of the junctional area. To assess the validity of the technique, a series of experiments were performed in mongrel dogs. Under cardiopulmonary bypass, the interatrial septum was incised along the tendon of Todaro, followed by cryosurgical ablation of the approach to the A-V node. Postoperative electrophysiological studies revealed complete conduction block between both atria and the A-V node. A spontaneous A-V nodal rhythm was produced. Initial activation was proved to be within the junctional area. This technique may lead to improved surgical management for atrial fibrillation and other supra-ventricular tachyarrhythmias. PMID- 8246349 TI - Effects of caffeine on ischemia-reperfusion injury in isolated rat hearts. AB - Cardiac sarcoplasmic reticulum (SR) plays an important role in regulation of the intracellular Ca2+ concentration. It is well known that intracellular Ca2+ overload is one cause of reperfusion injury. Thus, it is predicted that reperfusion injury of myocardium can be prevented by eliminating the Ca2+ overload. This study examined the effects of caffeine, a SR blocker, on reperfusion injury in isolated perfused rat hearts. Working hearts were reperfused for 25 min after 30 or 50 min of ischemia. Caffeine (10(-4) M) was administered during the period of ischemia or the initial 5 min of reperfusion. The left ventricular pressure and the electrocardiogram were recorded. Rate pressure products were calculated as an index of cardiac function. Adenine nucleotides were measured by high-performance liquid chromatography to assess energy charge. The administration of caffeine for a short period during the initial reperfusion significantly improved cardiac function in the hearts. Caffeine pretreatment during 50 min of ischemia, though, resulted in deterioration of both energy charge and cardiac function. Caffeine did not affect the incidence of either ventricular fibrillation or reversion to sinus rhythm. The energy charges were lower in the preparations with sustained ventricular fibrillation. PMID- 8246350 TI - Studies of cardiovascular responses to some endogenous pressor and hypotensive agents in conscious stroke-prone spontaneously hypertensive rats of different ages. AB - The effects of the endogenous pressor agents noradrenaline (NA), and angiotensin II (Ang II), and of the hypotensive agents acetylcholine (ACh) and adenosine (ADS), on blood pressure and heart rate in conscious and unrestrained stroke prone spontaneously hypertensive rats (SHR-SP) and normotensive Wistar Kyoto rats (WKY) of different ages (4-9 weeks old) were investigated. Pressor responses to NA were enhanced in 7- and 9- week-old SHR-SP compared with those in WKY, but pressor responses to Ang II in SHR-SP were not different from those in WKY at all ages. The bradycardias following pressor responses to NA and Ang II were markedly attenuated in SHR-SP, especially older ones. Hypotensive responses to ACH were enhanced in SHR-SP, particularly at 9 weeks of age. However, hypotensive responses to ADS were attenuated in SHR-SP, especially at 7 weeks of age. Transient fall of heart rate due to ADS was also attenuated in 7- and 9- week-old SHR-SP. These alterations of hemodynamic or cardiovascular responses in SHR-SP became more evident in the established stages of hypertension. These results suggest intimate relationships of the enhanced pressor responses to NA, attenuated bradycardias following pressor effects with NA or Ang II, and the attenuated hypotensive responses to ADS with the development or the maintenance of hypertension in SHR-SP. PMID- 8246351 TI - Influence of Abana on experimental atherogenesis in hypercholesterolemic rabbits. AB - An experimental, placebo-controlled study with rabbits was conducted to determine the role of Abana, an Ayurvedic preparation, on atherosclerosis. The results showed that Abana arrested the rise in TC, LDLc and VLDLc when administered along with an atherogenic diet for 60 days. In another phase of the study, when Abana was given from days 60-120 to rabbits who were made hyperlipidemic by feeding them an atherogenic diet for 60 days, Abana brought the TC, LDLc and VLDLc levels back to normal. On the other hand, the levels of the cardioprotective HDLc were elevated by Abana administration in both phases of the study. Histological pictures showed pronounced reduction in the atherosclerotic involvement of the coronary artery following treatment with Abana. The architecture of the coronary artery returned almost to normal. These findings substantiate the possible role of Abana in the reduction, prevention and reversal of atherosclerosis. PMID- 8246352 TI - [Percutaneous coronary angioplasty and levels of endothelin-1 in blood]. AB - The aim of this study was to assess plasma endothelin-1 (ET) concentrations during percutaneous transluminal coronary angioplasty (PTCA) of the left anterio descending coronary artery (LAD). We also examined relations between plasma concentrations of ET and diameter of the PTCA-balloon. Plasma levels of ET determined by radioimmunoassay were measured in 24 patients (pts) without previous myocardial infarction undergoing PTCA of LAD. The pts were divided into two groups. In group 1 (12 pts) plasma ET levels were measured in the right atrium and in the forearm vein 24 hours before PTCA, and 5 minutes after the last balloon deflation. In group 2 (12 pts) concentrations of ET were measured only in venous blood: 24 hours before PTCA, just before PTCA, 5 minutes after last balloon deflation and 24 hours later. We compared the diameter of the inflated balloon with the diameter of the dilated artery [balloon/artery ratio /b/a ratio/] and correlated it with plasma ET concentration rise in every case. [table: see text] The mean b/a ratio correlated with plasma ET concentrations after PTCA (r = 0.677 p < 0.001). CONCLUSIONS: Plasma ET concentrations do not rise after non complicated PTCA procedure. Because plasma ET levels depend on b/t ratio it is very important that the diameter of the dilating balloon should correspond with the diameter of the coronary artery to avoid overdilation. PMID- 8246353 TI - [Concurrent coronary artery disease and acquired rheumatic mitral valve disease]. AB - The aim of this study was to estimate coincidence of coronary artery disease (CAD) and rheumatic mitral valve disease in 264 patients treated in the National Institute of Cardiology (1976-1990). Severity of stenoses on coronary angiography with respect to age, sex, symptoms and risk factors were also estimated. Stenoses over 70% of artery lumen in relation to artery diameter before lesion and over 50% for left main coronary artery were stated as severe. There were 180 (68%) females and 84 (32%) males in the studied group; mean age was 52.5 year. Patients were divided into two groups: with angina--126 pts and without CAD symptoms--138 pts. 8 females (4%) had severe stenoses and 45 (25%) non-severe. Respectively 14 males (16.7%) had severe stenoses and 14 non-severe. Severe lesions were present in a group of females older than 50 years and in a group of males older than 45 years. Both in group with or without angina prevalence of coronary artery lesions was similar. Sensitivity and specificity of CAD clinical symptoms was low (less than 50%). Significantly more risk factors were present in pts with coronary stenoses than in pts free of CAD. No correlation between high pulmonary artery pressure and angina in patients without coronary stenoses occurred. PMID- 8246354 TI - [Pulmonary venous compliance. Indirect evaluation based on pulmonary capillary pressure registration]. AB - The compliance of pulmonary venous system (PV comp) is one of the most important factors influencing left ventricular filling pressure. The decrease of PV comp could be also one of the most important factors promoting secondary pulmonary hypertension. The aim of the study was to assess diagnostic value of PV comp based on some haemodynamic data in patients (pts) with isolated mitral stenosis (MS). PV comp was estimated according to Hirakawa equation (dV/dP = 0.4 * k2 x SV/(v - d), where k2 = 0.075 * PWP + 0.9) in 78 pts with MS, during preoperative right heart catheterisation with Vygon 5155-180 thermodilution catheters. The mean PV comp for the entire group of pts was 4.5 +/- 2.4 ml/mmHg. In pts with atrial fibrillation PV comp was significantly lower as compared with pts on sinus rhythm (3.7 +/- 1.9 vs 6.4 +/- 2.3, p < 0.0001. There was also a significant difference between NYHA I/II vs III/IV class as far as PV comp was concerned (5.1 +/- 2.3 vs 3.9 +/- 2.3, (p < 0.05). In the group with low PV comp less than 4 ml/mmHg pulmonary vascular resistance, RA, PA and amplitude of V-wave of PWP were significantly higher and stroke volume index of RV was lower than in the group with PV comp above 4 ml/mmHg. PWP did not differentiate the analysed groups. A decrease of PV comp is a sign of advanced mitral stenosis. PMID- 8246355 TI - [Unexplained syncope. Likelihood of inducing sustained ventricular tachycardia with electrophysiologic examination]. AB - In patients with unexplained syncope and documented nonsustained VT or complex premature ventricular beats indication for programmed ventricular stimulation (PVS) should be considered. However, the variables derived from noninvasive methods that predict diagnostic yield of PVS are incompletely defined. The purpose of this study was to elucidate the role of noninvasive testing variables in predicting inducible monomorphic sustained ventricular tachycardia (SVT) in 116 patients (pts) presenting with syncope. The study group consisted of 45 pts with remote Q wave myocardial infarction. 5 pts with non-Q wave myocardial infarction, 21 pts with another heart disease and 45 pts without organic heart disease. All pts underwent standard ECG, Holter monitoring, echocardiography, signal-averaged ECG and PVS with 1, 2 and 3 extrastimuli at three basic cycle lengths. Nonsustained VT, low ejection fraction, prolongation of QTc interval, abnormal Q waves in postinfarction pts (Q-MI) and late potentials (LP) indicate a greatly increased probability of inducible SVT. Inducible SVT was present in 15 of 45 (33%) pts with Q-MI. Although in the group of 71 pts without Q-MI, nonsustained VT on Holter monitoring was detected in 20 pts, LP in 16 pts, prolongation of QTc interval in 3 pts and low ejection fraction in 2 pts, the positive yield of PVS was documented in only one case (1%). The combination of Q MI and LP best predicted inducible SVT showing a sensitivity of 75%, specificity of 93%, overall predictive accuracy of 91%, predictive value of positive response of 63% and negative response of 69% to identify pts who would have inducible SVT. Thus, the combination of Q-MI and LP variables was shown as the best predictor of inducible SVT in pts with unexplained syncope. In pts without Q-MI the noninvasive testing variables have considerably less value in selection of pts to PVS, because inducible SVT is very rare in this population. PMID- 8246356 TI - [Constrictive pericarditis in a patient following prosthetic aortic valve replacement caused by postpericardiotomy syndrome]. AB - A case of a 53-year old man is presented who developed postpericardiotomy syndrome and subsequently constrictive pericarditis following prosthetic aortic valve replacement due to severe aortic stenosis. The diagnosis was based on clinical picture and confirmed invasively. Nearly total excision of thickened fibrous pericardium resulted in a slow but full disappearance of constriction signs. PMID- 8246357 TI - [Difficulties with pharmacologic treatment of a patient with atrioventricular nodal reentrant tachycardia]. AB - A case history of a 38 year old teacher with AVNRT is described. She had been treated for 11 years with 11 antiarrhythmic drugs in various combinations. No treatment prevented recurrence of arrhythmia. During long term treatment with class IA, IC, II, III and IV antiarrhythmic drugs, various side effects occurred. There was also suspicion of proarrhythmic effect, especially of prajmaline. Some of the drugs terminated tachycardia while administered intravenously. But often deep hypotonia, heart automatism disturbances and even asystole occurred, MAS syndrome occurred 5 times. The patient was referred to our laboratory to perform percutaneous radiofrequency ablation of the slow pathway. The procedure was performed without any complications. Efficacy of the ablation was proved by electrophysiologic study before and after intravenous atropine administration. During the 11 months follow-up the patient had no tachycardia. She is on no antiarrhythmic medication and continues her normal activity. PMID- 8246358 TI - [About riddles and threats during the ventricular repolarization period, in other words AT + U = ?]. PMID- 8246359 TI - [Why do countries differ in deaths from ischaemic heart disease?]. PMID- 8246360 TI - [Ventricular arrhythmias in apparently normal subjects]. PMID- 8246361 TI - Maintenance dialysis in North American children and adolescents: a preliminary report. North American Pediatric Renal Transplant Cooperative Study (NAPRTCS). AB - During 1992 the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) began to develop a pediatric Dialysis Patient Data Base by collecting data on pediatric patients who had received either hemodialysis (HD) or peritoneal dialysis (PD), or both, at a participating NAPRTCS center. This preliminary report describes study methods and contains detailed, though short term observations reported by 64 of 87 NAPRTCS centers on 762 patients who were < 21 years of age at enrollment and who received treatment between January 1, 1992 and September 15, 1992. In these 762 patients, a total of 810 independent courses of dialysis therapy were identified (PD = 534 [65.9%]; HD = 276 [34.1%]). Patients age groupings showed a significantly greater proportion of PD patients among younger age groups. Automated peritoneal dialysis was used by about 75% of registered PD patients at one and six months after registration. A total of 196 peritonitis episodes were reported, yielding a peritonitis rate of one episode every 7.1 patient-months. Ten percent of PD catheters were replaced, primarily for mechanical malfunction and leaks. Percutaneous catheters were used for vascular access in about one-half of the HD patients, with the remainder almost equally divided between arteriovenous fistulae and grafts. Vascular access revision was reported in 28% of HD patients, with about one-third of these revisions performed to create a more permanent access. Recombinant human erythropoietin therapy was used in 89% of PD and 94% of HD patients at six months. Recombinant human growth hormone therapy was used in 9% of PD and 5% of HD patients at six months.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246362 TI - Development of synergistic immunosuppressive drug regimens based upon cyclosporine. PMID- 8246363 TI - Five-year patient and graft survival in North American children: a report of the North American Pediatric Renal Transplant Cooperative Study. PMID- 8246364 TI - Five year survival of kidney transplantation in children: data from the European (EDTA-ERA) Registry. PMID- 8246365 TI - Metabolic effects of steroid withdrawal in adult renal transplant recipients. PMID- 8246366 TI - Prednisone withdrawal in HLA identical and one haplotype-matched live-related donor and cadaver renal transplant recipients. AB - Prednisone withdrawal was attempted in 121 of 915 renal transplants recipients between 1967 to 1992. There were 57 males, 64 females. Age range was 21 to 62 (mean 39.2 years). Etiology of renal failure was chronic glomerulonephritis (54), diabetic nephropathy (36), interstitial disease (17), hypertensive nephrosclerosis (6), and other (8). Kidney source was from HLA-identical living related donors (LRD) in 54 (Group I), one haplotype-matched LRD in 23 (Group II), and cadaver in 44 (Group III). Prior to the introduction of cyclosporin A (CsA) in 1984, prednisone withdrawal was attempted only in Group I. After 1984, prednisone withdrawal was also attempted in patients in Groups II and III, selected on the basis of having had no rejection episodes during the six months after transplantation. Forty-five patients in Group I were treated with azathioprine (Aza) and prednisone, and the remaining patients in Groups I to III were treated with Aza, prednisone and CsA. Mean follow-up was 93 months (6 to 207). Prednisone was gradually tapered and withdrawn in 94 of 121 patients after a mean period of 22.5 months (9 to 60). In 27 other patients, the prednisone dosage has been tapered to 5 mg/day or less with the aim of discontinuing the drug. There were seven episodes of acute rejection (3 during taper, and 1 each at 6, 7, 24 and 114 months after prednisone withdrawal); all seven were successfully reversed. Four other patients developed chronic vascular rejection (2 during taper and 2 after withdrawal).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246367 TI - Steroid withdrawal, rejection and the mixed lymphocyte reaction in children after renal transplantation. AB - With improved long-term graft survival after renal transplantation, cardiovascular mortality is emerging as the leading cause of death in adults and is also being reported in children. Chronic corticosteroid therapy is thought to be an important cause of post-transplant hyperlipidemia and hypertension. This study describes a steroid withdrawal protocol initiated to reduced cardiovascular risk factors in pediatric renal allograft recipients, reports on the rate of rejection observed and the use of an in vitro method to measure immunoresponsiveness and identify those patients who have not experienced a rejection episode. Thirty-six of 67 patients were able to discontinue prednisone and were maintained on cyclosporine alone. Twenty-two of the 36 patients experienced an acute rejection episode a mean of 14 months (range 1.5 to 36 months) after completion of the steroid taper. Ten of the 22 rejections occurred within 12 months after completion of the taper. Fourteen patients remain rejection free to date for a mean of 70.3 months (range 19 to 111 months) after withdrawal. Using the mixed lymphocyte culture reaction, we tested the hypothesis that steroid dependent recipients (SDR) will express donor antigen specific responsiveness and steroid independent recipients (SIR) will exhibit donor antigen-specific tolerance. Four of seven SDR were tolerant to donor specific antigens but responsive to unrelated controls, while five of six SIR were responsive to donor specific antigens. These unanticipated results highlight the complexity of allograft tolerance. PMID- 8246368 TI - Renal transplantation in the first five years of life. PMID- 8246369 TI - Management of hypertension after transplantation. AB - The multiple causes of hypertension in kidney transplant recipients complicate management. Most patients have at least two, if not more, reasons for elevated blood pressure. Determining the relative significance of these multiple causes is difficult. The kidney transplant population has a greater prevalence of "correctable" forms of hypertension than the general population. Even though the situation is complex, physicians should, therefore, proceed with a diagnostic assessment of the possible contributions of the native kidneys, vascular stenosis, chronic rejection, and drug therapy. It is important to consider transplant artery stenosis in recipients of pediatric kidneys or living-related donor kidneys. Both surgery and angioplasty for such lesions, however, are associated with the risk of allograft loss. Native kidney nephrectomy can control hypertension in some patients, but investigations that are specific and sensitive for this cause are lacking. Both cyclosporine and prednisone can cause hypertension. The higher the dose of either drug, the more likely they will cause hypertension. Hypertension alone is usually insufficient reason for discontinuation of either prednisone or cyclosporine. Medical management of hypertension, when no surgically correctable form has been found, relies on antihypertensive medication. Non-pharmacological measures (exercise, sodium restriction, etc.) can be expected to work as well as they do in the general population. Calcium channel blockers seem to preserve allograft blood flow better than other antihypertensive medications. Diuretics, while effective, may aggravate the lipid abnormalities of patients. PMID- 8246370 TI - Cyclosporine dosing and its relationship to outcome in pediatric renal transplantation. AB - The North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) contains information on more than 2,000 pediatric renal transplants performed since 1987. We reviewed the NAPRTCS database to determine the long-term trends in cyclosporine dosing for pediatric recipients of renal transplants and to determine whether there is a relationship between cyclosporine doses and graft outcome. More than 90% of all patients in the NAPRTCS registry received cyclosporine as initial therapy, with a slightly higher percentages of cadaver donor recipients than living-related donor recipients. Cyclosporine doses is all patients decreased from a median of 9 mg/kg/day at one month following transplant to 4.3 mg/kg/day at 36 months. Cyclosporine doses were generally higher in younger patients, particularly during the first year following transplantation. The relationships between cyclosporine doses and both rejection episodes and graft losses were assessed. Between 6 months and 36 months following transplantation, patients who had first rejection episodes or lost their grafts had lower median cyclosporine doses than those who did not. Proportional hazards regression analysis of the risk of having a first rejection episode more than one year after the transplant indicated that the cyclosporine dose was negatively correlated with risk of rejection (RR = 0.9, P = 0.02). We conclude that cyclosporine doses are routinely decreased following renal transplantation in children. Lower doses of cyclosporine are associated with rejection episodes, particularly late rejections. Cyclosporine doses of pediatric recipients of renal transplants should not be tapered. PMID- 8246371 TI - Long-term follow-up of growth in children post-transplantation. AB - With improving graft survival, growth post-transplantation, demands increasing attention from pediatric transplant physicians. As our data base matures, the North American Pediatric Renal Transplant Cooperative Study is able to track growth patterns longitudinally. Of 2086 patients enrolled in the registry, 412 patients have had a functioning graft for at least three years. The mean growth deficit for this group was -2.46 and the change in Z score at three years was 0.16 (P < 0.01). Of various factors that could conceivably impact on growth post transplantation such as donor source, race, number of transplants, and immunosuppressive therapy, only age at transplant and the original height deficit are significant. Most growth retarded children over the age of six years are unlikely to exhibit accelerated growth post-transplantation. PMID- 8246372 TI - Growth hormone in the treatment of growth failure in children after renal transplantation. AB - Growth failure continues to be a problem in the management of children with renal failure. Children have improved growth after successful renal transplantation, but seldom have "catch-up" growth, or achieve normal adult height. Several investigators have reported the use of rhGH in children after renal transplantation and demonstrated improved height, growth velocity, and standard deviation score (SDS). We review the existing literature, present additional data from our center, and offer possible explanations. PMID- 8246373 TI - Prednisone inhibits the efficacy of recombinant human growth hormone in pediatric renal transplant recipients. AB - To test the efficacy and toxicity of recombinant human growth hormone (rhGH, Protropin, Genentech), we reviewed our experience of its administration of rhGH to pediatric renal transplant recipients. Endogenous growth hormone (GH) levels were measured after stimulation with L-dopa and clonidine in growth retarded children whose height (ht) was greater than or equal to 2 standard deviations (SD) below the mean for age. Criteria for receiving rhGH were either subnormal GH levels (< 10 ng/ml) or a zero growth velocity over the preceding year despite normal GH levels after stimulation. The dose of rhGH administered subcutaneously was 0.1 mg/kg/day for 6 days/week. The efficacy of rhGH was evaluated using growth velocity index (GVI) and height standard deviation (Z) score. Catch-up growth was defined as an increase in Z score (delta Z) > or = 0.4. Twenty patients (17 with subnormal GH levels and 3 with normal GH levels but zero GVI) consented to rhGH treatment. Seventeen patients (14 males, 6 pubertal) have completed one year or more of rhGH therapy and are the subjects of this analysis. All 17 patients were receiving cyclosporine and 12 were receiving prednisone during rhGH therapy. Six of the 17 patients (35%) demonstrated catch-up growth (delta Z + 1.3 +/- 0.13). By regression analysis, the only factor noted to affect rhGH response was the concurrent use of prednisone. All five patients not receiving prednisone demonstrated catch-up growth compared to only 1 of 12 patients receiving prednisone (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246374 TI - Growth hormone treatment of growth failure among children with renal transplants. AB - Eight children with growth failure following renal transplant have been selected for recombinant human growth hormone (rhGH) treatment at Children's Hospital using the following criteria: (1) a functioning allograft for at least one year; (2) height < third percentile; (3) growth velocity < 4 cm/year; (4) growth potential; and (5) low-dose alternate-day glucocorticoid dosing. The children were 7.4 to 17.7 years of age; had received transplants 2.6 to 12.3 years before rhGH treatment; and all had decreased allograft function. The growth velocity of these children increased from 1.7 +/- 0.7 to 7.1 +/- 2.1 cm/year during the first year of rhGH treatment (0.05 mg/kg s.c. daily). The mean height SD score improved -3.9 +/- 1.5 to -3.4 +/- 1.3 in the seven children who completed one year of treatment (P < 0.001). There was no change in glucose tolerance during rhGH treatment. The serum creatinine concentration increased in all patients with a concomitant decrease in creatinine clearance. The mean rate of change in the inverse creatinine (1/Cr) increased from -0.005 +/- 0.004 dl/mg/month in the two years prior to rhGH treatment to -0.023 +/- 0.015 dl/mg/month during rhGH treatment (P < 0.01). The relative risks and benefits of rhGH treatment must be carefully considered for each patient. PMID- 8246375 TI - Predictors of growth response to rhGH in short children before and after renal transplantation. German Study Group for Growth Hormone Treatment in Chronic Renal Failure. AB - Many children with chronic renal failure (CRF) present with a reduced height and a reduced height velocity resulting in diminished final height irrespective of renal replacement therapy. Recombinant human growth hormone (rhGH) has become a new treatment modality for short renal patients, and the response to rhGH varies widely. In order to identify possible predictors of response to rhGH, the influence of sex, chronological age, bone age, pubertal status, height and height velocity at basal, target height, treatment modality for CRF, residual glomerular filtration rate (GFR), and steroid treatment was analyzed by single and multiple regression analysis in 49 children prior to and after renal transplantation. During the first treatment year with 28 to 30 IU rhGH/m2/week given by daily s.c. injections, height velocity was highest in patients on conservative treatment and lowest in patients on dialysis. Height velocity expressed in cm/year was inversely correlated with age (r = -0.63; P < 0.0001) and positively correlated with pretreatment height velocity (r = 0.65; P < 0.0001). The increment in height velocity SDS (chronological age) was significantly negatively correlated with the pretreatment height velocity SDS (chronological age) (r = -0.58, P < 0.001), indicating that at any given age the slowest growing children tended to respond best to rhGH treatment. It is concluded that the response to rhGH is significantly influenced by age, pretreatment height velocity, and treatment modality for CRF, whereas the influence of other variables is less important. PMID- 8246376 TI - Hospitalization in children during the first year after kidney transplantation. AB - Length of hospital stay post-renal transplant was investigated in 2171 North American pediatric patients. Hospitalization for those surviving one year with a functioning graft was 28.8 versus 36.0 days (P < 0.05) for living donor (LD) compared with cadaveric donor (CAD) recipients during the first year post transplant. Significantly prolonged hospital stays were recorded for LD recipients who (a) were less than one-year-old, (b) were receiving prophylactic ALG/OKT3, and (c) had a history of prior dialysis, and for CAD recipients who (a) were non-White and (b) received kidneys with cold ischemic times over 24 hours. In period II (2 to 6 months post-transplant), 51% and 68% of LD and CAD recipients, respectively were hospitalized while the corresponding values were 27% or 31% in period III (7 to 12 months post-transplant). Hospitalization was due mainly to graft loss or rejection episodes. Prolonged hospital stay coupled with poor graft survival might help to determine which aspects of the clinical practice of transplanting children warrant changes. PMID- 8246377 TI - Immunizations for pediatric transplant patients. AB - Infection remains a major cause of morbidity and mortality in transplant patients. Many infections, however, can be successfully prevented by immunization. This presentation reviews the problems associated with, and the questions that arise concerning the use of routine pediatric vaccines, such as diphtheria-pertussis-tetanus (DPT) and measles-mumps-rubella (MMR). It also reviews the use of special vaccines such as hepatitis B, pneumococcal, and influenza vaccines in transplant patients. Data concerning the use of two experimental, live-attenuated virus vaccines, against cytomegalovirus (CMV) and varicella, are discussed. The live-attenuated varicella vaccine can be predicted to decrease the morbidity and mortality of varicella-zoster virus infection in transplant patients. It has already been given successfully to immunocompromised children and is highly effective in the prevention of varicella. PMID- 8246378 TI - Recurrent disease in renal allografts. AB - Recurrent or de novo diseases account for only 5% of graft failure in children, but have much to teach us about mechanisms. In children, almost the only metabolic disease with recurrence is type I hyperoxaluria, in which the poor long term results of isolated renal transplantation makes combined liver and renal transplantation, or even prophylactic liver transplantation before renal failure the preferable alternative. While many forms of nephritis may show histological recurrence in allografts, it is notable that in many patients this is accompanied no clinical manifestations or only mild disease; this is particularly so in MCGN type II, IgA-associated nephropathy and Schonlein-Henoch purpura. However, FSGS and MCGN type I recur with frequency and severity. Hemolytic-uremic syndromes may recur, even repeatedly, but this is unusual. As many as 10% of pediatric grafts may show de novo membranous nephropathy, but in the majority this is mild or not clinically evident. In contrast, the rare anti-GBM nephritis affecting some patients with Alport's syndrome usually results in graft failure, but occurs in only a minority of Alport recipients. For all types of disease in allografts, risk factors for recurrence are poorly worked out, and attempts at treatment generally ineffective. PMID- 8246379 TI - Clinical significance of allograft glomerulopathy. AB - During the period from January 1973 to December 1990, 774 renal transplantations in 698 children were performed in our renal unit. A total of 540 grafts were examined both by light and immunofluorescence microscopy at least once. Recurrent glomerulonephritis was diagnosed in 62 grafts, de novo glomerulonephritis in 68 and allograft glomerulopathy in 38 (7%). The term allograft glomerulopathy (AGP) refers to the very specific pattern of glomerular involvement unique to renal allografts and characterized by: (1) widespread reduplication of the glomerular basement membrane due to a widening of the subendothelial space associated with (2) moderate increase in mesangial matrix and (3) interposition of mesangial matrix and cells. AGP is a glomerular manifestation of chronic rejection and is almost always associated with obliterative vascular changes. In our experience it is present in 25% of long-standing grafts (more than 10 years), but may develop early in the post-transplantation period (5 cases within the first year). In 13 of 33 patients in whom early biopsies were available, a different pattern characterized by a hypercellularity and the presence of monocytes-macrophages in the mesangial areas was observed. We suggest that this hypercellular pattern might be the early stage of the disease and could be called "acute allograft glomerulopathy" as opposed to the pattern characterized by reduplication of the glomerular basement membrane which deserves the name of "chronic allograft glomerulopathy." The exact relationship between these two patterns remains to be determined. Clinicopathologic correlations showed that chronic AGP is often associated with marked proteinuria. However, in our series proteinuria was absent or minimal in 25% of the cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246380 TI - Improving the utilization of cadaver kidneys in children. PMID- 8246381 TI - [Functional and morphological changes and liver regeneration after liver resection]. AB - The dynamics of functional and morphological changes and regeneration of the liver were followed-up in 192 out of 202 patients who underwent operation with resection of the liver for its various focal lesions. The main biochemical indices in the absence of cirrhosis are normalized in 6-7 weeks after extensive resection and in 3-4 weeks after economical resection. In cirrhosis the volume of the organ is not restored completely and normalization of the biochemical indices is essentially delayed. After extensive resection the volume of the remaining liver increases (up to 100%) in the early postoperative period (three weeks) with subsequent gradual diminution (to one year) to the volume of the noninvolved hepatic parenchyma measured before the operation. Regeneration of the liver is activated sharply in the early postoperative period. Study of these changes showed that adequate and stable compensation of the functional condition of the liver occurs in 12-18 months after extensive resection and in 2-4 months after economical resection. PMID- 8246382 TI - [Pancreonecrosis. Current views and perspectives]. AB - Generalization of data gained from the case records of patients with pancreonecrosis treated at 14 hospitals of Moscow allowed the authors to mark out typical errors in the diagnosis and therapeutic tactics which caused, to a considerable measure, mortality in 58.1 cases. From their personal experience in the treatment of such patients the authors believe it possible to improve essentially the diagnosis of acute pancreatitis by wide use of laparoscopy, ultrasonic scanning, and angiography of the pancreas for early verification of pancreonecrosis, appraisal of the condition of the extrahepatic bile ducts, and application of mildly injurious methods of treatment. Setting apart only two forms of acute pancreatitis is substantiated from the clinico-anatomical standpoint. The choice of the therapeutic tactics in pancreonecrosis should be based on the severity of the disease and the dynamics of its development during intensive therapy, rather than on the morphological diagnosis. Using the criteria of evaluation of the severity of pancreonecrosis, the authors reduced considerably the frequency of operative interventions in the early stages of the disease. Among the methods of operative treatment of pancreonecrosis, distant pancreatectomy or programmed repeated necroso-sequestrectomies are preferred. PMID- 8246383 TI - [Laparoscopic cholecystectomy]. AB - The techniques of laparoscopic cholecystectomy is described. The operation was carried out on 37 patients with various diseases of the gallbladder. The indications and contraindications for this operation are discussed. The authors did not encounter serious postoperative complications. Infiltration developed in the region of the umbilical incision in one case. All patients were discharged from the clinic on the 2nd to 11th day. The authors believe expedient the organization of special centers for training in laparoscopic cholecystectomy. PMID- 8246384 TI - [Benign cystic tumors of the pancreas]. AB - Benign pancreatic cystic tumors (BPCT) is a rare and little studied pathological condition. By the end of the eighties approximately 500 registered cases of BPCT were described in the world literature, which were those of different morphological variants of cystadenomas (CA). Many scientists note the high probability of malignant degeneration of some CA types. The diagnostic signs and surgical tactics in BPCT are treated ambiguously. The authors systematized the most characteristic signs of pancreatic CA and revealed the possibility of accurate preoperative recognition of these tumors. From rich clinical material (21 cases) the authors show that complications resulting from an erroneous choice of the operation are inevitable. Postoperative complications occurred in 8 patients. The late-term results of the operation were followed-up in 11 patients and were found to be good and satisfactory in complete removal of the tumor. According to foreign authors, cystic carcinomas of the pancreas account for 1-2% ot all primary carcinomas of the organ. Timely radical operation dictated by the specific features of pancreatic cystadenoma is one of the measures of carcinoma prevention. PMID- 8246385 TI - [Aortic valvuloplasty in acquired valvular disease]. AB - On the basis of the results of studying the morphology, biomechanics, and physiology of the aortic root and the cusps of the aortic valve the authors suggest a substantiated set of surgical manipulations which allow the patient's natural valve to be preserved. Coarse calcinosis of the cusps and fibrous ring of the aortic root is the only absolute contraindication for valvuloplasty. The authors' clinical experience consists of the first 100 operations. Hospital mortality--8.0%. With intraoperative appraisal of hemodynamics, good correction of the abnormality was achieved practically in all patients. The maximal follow up period by April, 1991 was 5.5 years. During that period four patients were again subjected to operation because aortic insufficiency recurred. The authors recommend their experience with plastic operations on the aortic valve for wide clinical application. PMID- 8246386 TI - [Neuroimmune correlations analysis in choosing the surgical method for treatment of patients with bronchial asthma]. AB - The activity of vegetative reception, immunological reactivity, and glucocorticoid provision was studied in 196 patients with bronchial asthma who were admitted for operative treatment. The authors devised the principle of the stage-by-stage impairment of the activity of vegetative reception, immunological reactivity, and glucocorticoid provision during the formation of bronchial asthma and distinguished six types of neuroimmune interrelations. On the basis of these six types they suggest pathogenetically substantiated indications for various methods of surgical treatment of patients with bronchial asthma. PMID- 8246387 TI - [Surgical treatment of local recurrence of malignant lung neoplasms]. AB - Forty-seven reoperations were performed on 46 patients for local recurrence of carcinoma and other malignant tumors of the lung after surgical treatment. In two thirds of cases the local recurrence was a consequence of various errors in therapeutic tactics and surgical techniques made during the operation for the primary tumor. The malignant lung tumor recurred in periods of 4 weeks to 20 years after the operation, but mostly (in 74.5% of patients) in the first 5 years. The authors believe that the criteria of a local recurrence are the same morphological structure as of the primary tumor and localization of the second tumor in the stump of the resected bronchus or in the zone of the previous operation. In 17.0% of cases the recurrent tumor was less differentiated than the primary tumor, and in 34.0% it metastasized to the regional lymph nodes. Pneumonectomy-type removal of the remaining parts of the pulmonary tissue was the operation of choice in local recurrence of lung carcinoma. Organ preserving operations were performed only in local recurrence of less malignant tumors. Five year survival after repeated operations in local recurrence of malignant lung tumors was 42.6% and depended on the morphological structure of the recurrent tumor. PMID- 8246388 TI - [Removal of esophagus using a one-stage repair]. AB - The article deals with the results of operations performed on 306 patients for extirpation of the esophagus and one-stage total esophagoplasty with the formation of the anastomosis on the neck. The operation was conducted on 178 patients with carcinoma and 128 patients with benign esophageal strictures. The authors claim this operation to be the operation of choice in esophageal carcinoma, cardioesophageal carcinoma, burn and peptic strictures, and in patients with stage IV cardiospasm. An isoperistaltic gastric tube or the large intestine is used for replacement of the esophagus. The postoperative mortality rate was 3.3%. The immediate and late-term results of this operation are much better than those of the other types of esophagoplasty. The authors recommend the operation to be used widely in the practice of specialized clinics for the management of the above mentioned diseases. PMID- 8246389 TI - [Clinical aspects and therapy of cholelithiasis in elderly patients]. AB - The article analyses the results of surgical treatment of 110 patients aged over 70 with acute cholecystitis. All of them underwent cholecystectomy with intraoperative cholangiography: choledocholithiasis was found in 26.4% and stricture of the distal choledochus in 10.0% of cases. Postoperative complications developed in 19% of patients, the mortality rate was 5.45%. Simultaneously, 72 elderly and old-aged patients were operated on for chronic calculous cholecystitis after thorough examination and preoperative management. Postoperative complications occurred in 13.9% of cases, none of the patients died. The authors conclude that in adequate preoperative management, careful choice of the method of anesthesia, and substantiated postoperative management, cholecystectomy is the operation of choice in acute calculous cholecystitis in patients over 70 years of age. Planned operations are very advisable in chronic calculous cholecystitis in patients of this age group for the prevention of severe complications, if adequate treatment of concomitant diseases is applied. PMID- 8246390 TI - [Methods for increasing graft survival in esophagogastroplasty]. AB - From their own clinical material the authors revealed that insufficient venous outflow was the main link in the pathogenesis of impaired viability of the pedicle formed from the greater curvature in esophagoplasty. This leads to circulatory disorders in the graft: venous hypertension of up to 34.4 +/- 2.3 cm inadequate perfusion of tissue, and secondary arterial hypotension below 40 mm Hg. The authors suggested an original pathogenetically substantiated method of esophagoplasty with a pedicle from the greater curvature of the stomach. Its essence consists in the creation of an additional venous outflow from the graft into the neck veins. Operations were performed on 55 patients with the formation of 70 intravenous anastomoses. The suggested method made it possible to form a longer graft and reduce considerably the number of necrotic complications in esophagoplasty. A reliable graft of sufficient length allowed the immediate and late-term results of treatment and rehabilitation of patients with esophageal carcinoma to be improved essentially. PMID- 8246391 TI - [Intraoperative ultrasonography in Itsenko-Cushing's disease and syndrome]. AB - Ultrasonic study conducted during an operation is finding wider use lately. Surgery is the main method of treatment of most diseases of the adrenals, therefore problems of topical intraoperative diagnosis are very urgent. Twenty three patients were operated on for Itsenko-Cushing's disease and syndrome and tumors of the retroperitoneal space at the Faculty Surgical Clinic of the Moscow Medical Academy within a period of 12 months. All patients were subjected to complex ultrasonic examination, including pre-intra- and postoperative study. Intraoperative ultrasonic study was conducted in two stages: stage 1--examination after thoracotomy through the diaphragm for choosing the optimal place and length of phrenotomy; stage 2--ultrasonic examination after phrenotomy to localize precisely the adrenal or tumor and reveal their connection with the adjacent organs and vascular structures. Complex ultrasonic examination has advantages over the other examination methods (computed tomography, angiography, scintigraphy) which allows it to come to the forefront in the diagnostic search for adrenal tumors. Intraoperative ultrasonic examination makes it possible for the surgeon to avoid wide phrenotomy and prolonged investigation, reduces the possibility of inflicting injury to the large vessels, which reduces the duration of the operation and anesthesia, and improves the course of the postoperative period. PMID- 8246392 TI - [Role of osteoscintigraphy in postoperative monitoring of bone graft survival]. AB - Osteoscintigraphy was conducted in three patients in the immediate and late-term period after transplantation of allobrephobone with revascularization in the recipient zone. The method is compared with the results of immunological monitoring and histological control. Radionuclide study in combination with other methods allowed judgement of the adequacy of blood circulation in the bone allobrephotransplant and its viability. It is suggested that osteoscintigraphy of the transplant in comparison with the corresponding intact segments of the skeleton may serves as a criterion for this transplant in various postoperative periods. PMID- 8246393 TI - [Successful surgical treatment of massive thromboembolism of the pulmonary artery]. PMID- 8246394 TI - [Organ xenotransplantation (Review of foreign literature)]. PMID- 8246395 TI - [Studies on oxygen content in aqueous humor of the anterior chamber of the rabbit's eye--I. Oxygen content in aqueous humor of anterior chamber and arterial blood under physiologic conditions]. AB - The measurements of the partial pressure of oxygen pO2 in the aqueous humor of anterior chamber in both eyes and arterial blood, using the electrochemical- polarographic method, under physiological conditions, have been carried out in 20 rabbits. The mean value of pO2 oxygen partial pressure in aqueous humor was 8.48 kPa, whereas in arterial blood was higher--9.69 kPa. The processes of infiltration (diffusion) from blood-vascular system and oxygen distribution in the aqueous humor of anterior chamber of the eye have been discussed. PMID- 8246396 TI - [Studies on oxygen content of aqueous humor in the anterior chamber of rabbit's eye--II. Oxygen content of aqueous humor in anterior chamber and arterial blood in experimentally provoked ischemia]. AB - The ligature of the right common carotid artery has been performed in 20 rabbits. Then, the partial pressure of oxygen pO2 in the aqueous humor of anterior chamber of the both eyes and in the right and left ear artery was measured. An electrochemical polarographic oxymeter was used for this purpose. The measurements were carried out on the second day, in a week and in one month after performing the ligature. A statistically significant decrease of pO2 in aqueous humor of anterior chamber of the right eye and of ear artery on the right side was observed, especially on the second day and after a week. Blood circulation stoppage within the right common carotid artery causes, in spite of the preservation of collateral circulation, a significant pO2 decrease in aqueous humor. The pO2 measurement would be a valuable supplement to the methods applied in the measurement of ear artery blood circulation. PMID- 8246397 TI - [Tumors of the eye in childhood]. AB - Cases of benign and malignant tumors treated in Children's Ophthalmological Clinic of Silesian Medical Academy in Katowice in the years 1982-1991 were presented. The clinical material comprises 135 eyes in 129 children. The benign tumors occurred in 83 children (64.3%) and malignant tumors in 46 children (35.7%). The most often observed benign tumor was naevus pigmentosus (23.7%) and the malignant tumor was mainly retinoblastoma (34.8%). PMID- 8246398 TI - [Results of treating malignant conjunctival melanoma]. AB - 28 patients with histologically confirmed conjunctival melanoma were treated between 1968 and 1987. In all the cases, the lesion was excised and in 15 cases, it was secondarily irradiated with 60Co or 27Sr. In 9 cases, recurrence appeared and, among them, twice as often in the group treated only surgically. 6 patients with the recurrence of conjunctival melanoma died because of metastases, 2 are alive and there is no information about 1. PMID- 8246399 TI - [Birdshot chorioretinopathy]. AB - The clinical picture of birdshot chorioretinopathy has been presented, basing on the studies of 102 cases, treated in 16 university clinics and ophthalmological departments in Europe. The records of the patients were sent to the Ophthalmological Clinic of Ghent University as a contribution to an international research programme concerning birdshot chorioretinopathy. The observations reveal that this type of chorioretinopathy is a separate entity and its diagnosis should be based on the binocular occurrence of characteristic lesions in choroid, accompanied by at least two of the following symptoms: vitreous inflammation, vasculitis, macular edema and papilledema. PMID- 8246400 TI - [Clinical description of pseudoexfoliation of the lens capsule]. AB - The authors presented their clinical observation of 75 cases of pseudoexfoliation syndrome. The most frequent reasons for admission were glaucoma (65%) and cataract (27%). The diagnosis of pseudoexfoliation of lens capsule was made basing on the examination of the anterior segment of the eyeball with slit lamp. Glaucoma that occurs with the pseudoexfoliation of lens capsule is of secondary, open angle nature. PMID- 8246401 TI - [Evaluation of visual acuity of small children with preferential looking methods]. AB - With preferential looking (PL) techniques, especially the Acuity Card Procedure (Teller Acuity Cards), the visual acuity in small children can be determined. PL testing was used in 294 children in the Strabismus and Amblyopia Department of the Ophthamology Hospital in Krakow. These children had been directed from other ophthalmologic departments. Almost all of them showed general retardation and their visual acuity could not be tested with traditional methods. The testing was considered positive when the visual acuity of each eye could be determined separately. Such result was obtained in 230 children (78.2% of all patients). The remaining group consisted of 59 children (20.0%) who were tested only binocularly and 5 children (1.8%) by whom no testing was possible. PMID- 8246402 TI - [Ocular symptoms in Wegener's granulomatosis--personal observations]. AB - The authors present 10 patients treated for Wegener's granulomatosis. In 5 of of them (50%) the ocular changes were observed: in 3-infiltration of the orbit, including one case with infiltration of lacrimal ducts, and in 2-the recurrent bacterial conjunctivitis. On the account of the rareness of the disease and its various symptoms, the diagnosis, in some cases, was made fairly late. It was emphasized that the ocular lesions may be the first symptoms of Wegener's granulomatosis. The bacterial conjunctivitis, keratitis, scleritis, and orbital pseudotumor in patients with chronic respiratory tract and nasal sinuses changes should arouse suspicion of Wegener's granulomatosis and are an indication for detailed tests. PMID- 8246403 TI - [Treatment of orbital hemangioma]. AB - Hemangiomas occur in 1%-2.6% of newborn infants and, after 7 years, they disappear in 70% of them. Indications for the treatment are the deterioration of visual function and great cosmetic defects. Both surgical and radiological therapy are likely to cause numerous complications. The reports concerning the beneficial effect of corticosteroids, injected directly into hemangiomas, have recently been presented. In two of our patients, aged 32 and 43, the injection of steroids resulted in the disappearance of hemangioma during 2-4 weeks. PMID- 8246404 TI - [Local use of cyclosporin in nonbacterial corneal ulceration]. AB - The authors present the results of local use of cyclosporin solution in 6 patients with corneal ulcerations occurring in Sjogren disease, rheumatic disease and after corneal transplantation. The ulceration was healed in all patients. The cyclosporin, locally applied, has no side effects and may be an efficient way in the treatment of some immunological diseases of cornea. PMID- 8246405 TI - [Morgagni-Steward-Morel syndrome accompanied by venous stasis on the edge of the optic disc]. AB - A case of a 41-year-old woman with Morgagni-Steward-Morel syndrome is presented. The first symptom of the disease was papilledema. The results of clinical examination and additional tests eliminated the presence of intracranial tumor and revealed the hypertrophy of the internal laminal of frontal bone. The patient also suffered from glaucoma, hormonal disorders and considerable obesity. Both the clinical picture and the results of laboratory and radiological tests permitted to diagnose Morgagni-Steaward-Morel syndrome. PMID- 8246406 TI - [Abuse of local anesthetics in ophthalmology]. AB - Surface anaesthesia is applied in ophthalmology for minor surgery, diagnostics tests and also in cases of corneal or conjunctival inflammation and injuries. The abuse of such anaesthetics may cause serious complications. The authors emphasize the necessity for their careful application. PMID- 8246407 TI - [Ticlid : clinical use and current concepts about the mechanism of action]. PMID- 8246408 TI - [Transthyretin. Physical and chemical properties and biological role in the human body]. PMID- 8246409 TI - [Differential use of vasodilators in various types of cardiac insufficiency]. AB - Vasodilators (captopril or nifedipine) were administered to 85 patients with stage IIA-IIB circulatory insufficiency. Of them 36 had dilated cardiomyopathy (DC), 30 ischemic heart disease (IHD), 19 suffered from essential hypertension (EH). Morphofunctionally, myocardial lesion was of a dilated type in 50 patients (36 DC, 10 IHD and 4 EH cases), of a hypertrophic type in 18 patients (4 IHD, 14 EH cases), of a mixed type in 17 IHD patients. The response to acute tests with 25 mg captopril and 20 mg nifedipine along with clinicohemodynamic results of 3-5 week course treatment with the drugs point to noticeable advantage of captopril in the dilated lesion (18 responders of 26 patients), while nifedipine in the hypertrophic type (10 responders of 10 patients). In mixed-type IHD patients both drugs displayed high efficacy. PMID- 8246410 TI - [Latent cytomegalovirus infection, apolipoproteins B and A-1 in blood plasma and atherogenesis]. PMID- 8246411 TI - [Psychasthenia - clinical variations of psychopathy]. PMID- 8246412 TI - [Valvular prolapse (clinical, echocardiographic, phonocardiographic and electrocardiographic features)]. AB - Clinical, echoCG, PCG and ECG examinations were performed in 42 patients with primary isolated valve prolapses: mitral, aortal and tricuspid (32, 5 and 5 patients, respectively). It is shown that degree 1 regurgitation caused no hemodynamic disturbances, that of degree 2 and 3 brought about cardial complaints in more than half the patients, hemodynamic overstrain of the heart, sound symptoms similar to those of relevant valvular disease. PMID- 8246413 TI - [Myocardial infarction in systemic lupus erythematosus]. AB - Six patients under 50 with confirmed systemic lupus erythematosus (SLE) developed myocardial infarction at different intervals from the disease onset (1-21 years). Five of them had chronic disease, one suffered from acute SLE. The analysis of clinical findings identified myocardial infarction risk factors in SLE (chronic course of the disease, long-term glucocorticosteroid treatment, hypercholesterolemia, persistent nephrotic syndrome, arterial hypertension, vasculitis, hypercoagulation, antiphospholipid antigens) and provided a basis for prevention of this severe SLE complication. PMID- 8246414 TI - [Difficulties and improvements in the diagnosis of early gastric cancer]. PMID- 8246415 TI - [Monocyte aggregation as a criterion in the evaluation of mononuclear phagocytes function in patients with bronchial asthma]. AB - The study was made of arachidonate-induced aggregation of peripheral blood monocytes in 18 patients with bronchial asthma versus 12 controls. Monocytic aggregation and dependence of the patients on glucocorticosteroids appeared related: in hormones untreated patients the aggregation was more active than in controls. The aggregation of hormone-dependent patients and controls showed no significant differences. Allergic reaction correlated in severity to aggregation intensity. The method of graphic registration of arachidonate-induced aggregation of peripheral blood monocytes is promising in evaluation of immunity macrophagal monocytic component in asthmatic patients. PMID- 8246416 TI - [A case of angioimmunoblastic lymphadenopathy]. PMID- 8246417 TI - [A case of primary hyperparathyroidism complicated by disseminated joint disease]. PMID- 8246418 TI - [Pellagra as an early manifestation of Crohn disease]. PMID- 8246419 TI - [Effectiveness of trobicin (spectinomycin) effectiveness in acute gonorrhea of men]. PMID- 8246420 TI - [Syndrome X - a special form of ischemic heart disease?]. PMID- 8246421 TI - [Diagnosis and treatment methods of osteoarthrosis]. PMID- 8246422 TI - [Treatment of patient with cardiac fibrillation]. PMID- 8246423 TI - [Croupous pneumonia: current and unresolved aspects]. PMID- 8246424 TI - [Requirements for the physician's report and its presentation to the certifying committee for certification by specialty boards]. PMID- 8246425 TI - [Various conventional views on ulcer patients- are they real?]. PMID- 8246426 TI - [Problems of battle-field surgery in the "Clinical Medicine" journal]. PMID- 8246427 TI - [Certain situations may foster intrigue. Intrigue is part of working life]. PMID- 8246428 TI - [Job hunting during unemployment. II. Unemployment insurance]. PMID- 8246429 TI - [The right to request]. PMID- 8246430 TI - [Why Mrs. Weber had a reputation of being difficult]. PMID- 8246431 TI - [Psychological care of patients with kidney disorders--experiences. Life with dialysis]. PMID- 8246432 TI - [Important decisions--inferior organization]. PMID- 8246433 TI - [Delegated nursing care in psychosocial medicine]. PMID- 8246434 TI - [Getting rid of lice...]. PMID- 8246435 TI - [An instrument for the evaluation of quality--the model of the Swiss Nursing Association. Measuring quality? It is possible]. PMID- 8246436 TI - [The quality of care given to patient A]. PMID- 8246437 TI - [Keeping the aged with psychiatric disorders at home. The day care center]. PMID- 8246438 TI - [Stop those false suggestions]. PMID- 8246439 TI - [Return from Albania]. PMID- 8246440 TI - [Preserving the earth which allows us to live. The threatened soil]. PMID- 8246441 TI - [Part 1: How do decubitus ulcers start?]. PMID- 8246442 TI - Nephritogenic autoantibodies in systemic lupus erythematosus: immunochemical properties, mechanisms of immune deposition, and genetic origins. PMID- 8246443 TI - Synergistic effects of vascular endothelial growth factor and basic fibroblast growth factor on the proliferation and cord formation of bovine capillary endothelial cells within collagen gels. AB - BACKGROUND: Angiogenesis occurs within the interstitial matrix. We therefore cultured endothelial cells within collagen gels rather than on the surface of culture dishes. We measured the effect of a human glioblastoma-derived factor, and the combined effect of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), on both the proliferative and morphologic changes exhibited by endothelial cells. EXPERIMENTAL DESIGN: Bovine capillary endothelial (BCE) cells were cultured in a gel of type I collagen. Effects of growth factors were evaluated by the cell recovery from the gel and the morphologic changes of cells. The glioblastoma-derived factor was characterized by column chromatography, immunoblotting, and immunoadsorption with an anti-VEGF antibody. RESULTS: After 4 days of culture in the collagen gel with 10% calf serum Dulbecco's minimum essential medium, most of the BCE cells died. Addition of glioblastoma-conditioned medium resulted in a change in morphology from a round shape to an elongated spindle shape; moreover, the number of BCE cells was enhanced. The major activity in the conditioned medium was VEGF. In the 3 dimensional gel, we found a higher activity associated with VEGF, compared with bFGF when the growth factors were added 24 hours after seeding. In this assay, the response to bFGF diminished within 24 hours, but that to VEGF continued for 48 hours. When the two factors were added to the culture simultaneously, the cell number was greater than the sum of those stimulated with either growth factor alone. In the presence of both bFGF and VEGF, cord-like structures became prominent in the BCE cell cultures. CONCLUSIONS: A human glioblastoma cell line produced VEGF, which enhanced the proliferation of BCE cells and induced an elongated shape in collagen gels. VEGF and bFGF increased the rate of proliferation and the number of cord-like structures in a synergistic manner. PMID- 8246444 TI - Differentiation of pancreatic acinar cells into duct-like cells in vitro. AB - BACKGROUND: Previous studies have shown that a combination of both the extracellular matrix and secretagogues plays critical roles in the maintenance of well-differentiated pancreatic acinar cells in culture. In the present study, we have shown that, upon proper stimulation, acinar cells change their growth pattern and morphologic appearance to a duct like phenotype. EXPERIMENTAL DESIGN: Both rat and guinea pig acinar cells were cultured on or embedded into the Matrigel basement membrane, in the presence of differentiating agents such as dimethylsulfoxide (DMSO), hexamethyl-bis-acetamide, dimethylformamide, triiodothyronine, and butyric acid. The growth patterns, cell proliferation, ultrastructural appearance, intracellular contents, secretion and immunolocalization of amylase, as well as the expression of the ductal marker carbonic anhydrase II and lectin-binding specific sites were analyzed. Moreover, the effects of metabolic inhibitors such as cycloheximide and actinomycin D on the DMSO induced action were also examined. RESULTS: Isolated acinar cells from both rat and guinea pig pancreas showed an important modification of their growth pattern and morphologic appearance when culture embedded into Matrigel in the presence of 2% DMSO. They reaggregate and form isolated branched tubular structures lined by a single cell layer. These tubules can be maintained in culture for over a period of 21 days. The cells lining the tubules were originally acinar cells that became elongated and progressively lost their secretory granules. They displayed a lower number of apical microvilli and established long junctional complexes with elaborated interdigitations. The immunocytochemical localization and biochemical determination of intracellular and secreted amylase revealed a progressive decrease reaching minimal values by the 12th day of culture. The cells further expressed the duct cell marker carbonic anhydrase II and lost the Helix pomatia lectin-binding affinity characteristic of acinar cells. Cell proliferation by modified cells as measured by thymidine incorporation and the autoradiographic labeling index, was significantly lower than in control cultured acinar cells. The DMSO differentiating action was mimicked, but to a lesser extent, by the other agents except butyric acid. Since cycloheximide and actinomycin D inhibited the DMSO induced changes, protein synthesis and DNA transcription seem to be required. CONCLUSIONS: Our results demonstrate that normal pancreatic acinar cells retain a morphogenetic plasticity and, upon particular stimulation, can change their differentiation commitment pattern toward that of the duct cell phenotype. PMID- 8246445 TI - Glomerulonephritis induced by human IgMK-IgG cryoglobulins in mice. AB - BACKGROUND: Human cryoglobulinemia is sometimes associated with glomerulonephritis (GN) due to deposition of cryoglobulins (cryos). To see whether human cryos can induce GN in mice and to study time-related changes of glomerular lesions and possible factors of cryos' nephritogenicity, we developed an experimental passive model of cryoglobulinemic GN. EXPERIMENTAL DESIGN: Two cryos IgMk-IgG from 2 patients with active GN (OLD and SOR), 2 cryos IgMk-IgG (TAC and GRO) and 1 IgM lambda (CHI) from 3 patients without GN were purified, solubilized at 37 degrees C and injected intravenously into BALB/c mice, 4 mg, twice a day. To study the possible factors of cryo nephritogenicity, we analyzed: (a) the presence, amount, and size of complexed IgMk-IgG at 37 degrees C by fast flow liquid chromatography; (b) the Cc1 or Lc1 subclass of rheumatoid factors; (c) the isoelectric points of the IgMks; (d) The proportion of IgG subclasses in cryos. RESULTS: On day 1 from the beginning of intravenous injections, cryos OLD had induced mesangial deposits of human IgM, human IgG, mouse C3 and mesangial hypercellularity. On day 2, phagocytizing cells were found along with massive endoluminal and subendothelial deposits of IgM, IgG, and C3. On day 6, perivascular infiltrates of mononuclear cells were also seen. Cryos SOR induced a similar but milder form of GN. After administration of purified OLD IgMk, OLD IgG, GRO IgMk or GRO IgG, only OLD IgMk was deposited in the mesangium. Analysis of all the cryos revealed that: the amount of complexed IgMk-IgG at 37 degrees C was always less than 1% of cryos; Cc1 and Lc1 idiotypes were not related to the nephritogenicity of cryos, the isoelectric points of IgMks were 4.5 to 5.5 and IgG1 was the prevalent subclass. CONCLUSIONS: Data demonstrate that human cryos from patients with GN can induce GN in mice that resembles the corresponding human pathology. The affinity of IgMk for glomeruli and the unexpectedly small amounts of IgMk-IgG complexes at 37 degrees C suggest that there is a role of in situ binding in nephritogenicity which is independent of the isoelectric point, rheumatoid factor idiotype, or IgG subclass. PMID- 8246446 TI - Location and alternative splicing of type XIII collagen RNA in the early human placenta. AB - BACKGROUND: Type XIII collagen is widely distributed in the fetus. It is characterized by complex alternative splicing of its primary transcripts in regions corresponding to nine exons of the gene. EXPERIMENTAL DESIGN: Localization of type XIII collagen mRNAs in early placentas was determined by in situ hybridization. Reverse-transcription-polymerase chain reactions were used to examine alternative splicing of nine exons in villous and decidual mRNAs. RESULTS: An intense in situ hybridization signal was observed in the fibroblastoid stromal cells of the placental villi. A moderate signal was found in the endothelial cells of developing capillaries and the cells of the cytotrophoblastic columns. Furthermore, mRNAs were detected in the large decidual cells of the decidual membrane and the stromal cells of the gestational endometrium, but not in the epithelial cells in the endometrial glands. Five combinations of exons 3B, 4A, 4B and 5, encoding half of the COL1 domain, were found. The combination lacking exons 3B-5 was the major variant in both villous and decidual mRNAs. Three combinations of exons 12 and 13 encoding the NC2 domain were found, the long variant containing either 12 or 13 sequences being the major variant in the villi while nearly equal amounts of long and short variants lacking both 12 and 13 sequences were observed in the decidua. Four variants of exons 29, 33 and 37, encoding parts of COL3 and NC4, were found as splicing out of exon 37 was not detected. The major variants in both mRNAs were two that lacked exon 29 and either lacked or contained exon 33 sequences. CONCLUSIONS: Type XIII collagen mRNAs were located in the placenta. Due to alternative splicing, the lengths of the COL1 and NC2 domains of this collagen vary from 57 to 104 and from 12 to 34 amino acids, respectively. The COL3 domain varies between 208 and 235 residues, whereas the NC4 is 18 residues. PMID- 8246447 TI - Ultrastructural localization of heparin to human mast cells of the MCTC and MCT types by labeling with antithrombin III-gold. AB - BACKGROUND: Mast cells derived from human skin and lung have been reported to produce heparin and chondroitin sulfate E proteoglycans. However, no information about the proteoglycans distribution among the different human mast cell types (MCTC and MCT) is available. Conjugates of antithrombin III-gold were used to assess the presence of heparin in both human mast cell subsets. EXPERIMENTAL DESIGN: Thin sections of human and rodent tissues and dispersed cell preparations were labeled with the conjugate in the presence of saline, heparin, and chondroitin sulfates A and E and particle densities were measured over granules, perigranular regions, and extracellular space. Control sections were preincubated with heparinase, chondroitinase ABC, or buffer. RESULTS: Labeling with antithrombin III-gold particles was detected in essentially all granules of human mast cells in skin (predominantly MCTC type), lung alveolar wall, and bowel mucosa (predominantly MCT type), but was negligible over human eosinophils. Consistent with the known distribution of heparin in rodent mast cells, strong labeling was observed over rat peritoneal connective tissue type mast cells, but not over mucosal mast cells in bowel mucosa of Nippostrongylus brasiliensis infected rats (which contain chondroitin sulfate di-B) nor over mouse PT-18 mast cells (which contain chondroitin sulfate E). Mast cell labeling was preferentially blocked by exogenous heparin, and virtually abolished by heparinase but not chondroitinase ABC preincubation. CONCLUSIONS: The data with rodent mast cells indicate that antithrombin III-gold labels cells that contain heparin, but not those that contain only over-sulfated chondroitin sulfates. Specificity of the procedure for detecting heparin is further demonstrated by inhibition of labeling after preincubation with heparinase and by competition with exogenous heparin. On this basis, we conclude that heparin is present in essentially all mast cells in normal skin, lung alveolar wall, and bowel mucosa. The presence of heparin in all human mast cells is different than for rodent mast cells, and probably accounts for the inability to clearly distinguish different human mast cell types from one another with histochemical stains based on proteoglycan content. PMID- 8246448 TI - Immunoreactivity of the nuclear antigen p105 is associated with plaques and tangles in Alzheimer's disease. AB - BACKGROUND: The protein p105 is a nuclear antigen associated with cell proliferation. Recent investigations have indicated that many molecules previously considered to be growth-associated are actually involved in plasticity of the central nervous system. EXPERIMENTAL DESIGN: For the present study, using immunohistochemical and immunochemical techniques in control and Alzheimer's disease (AD) brain, we explored the possibility that p105 could be involved in the aberrant sprouting response. AD tissue was used because in this disorder, the neuronal sprouting processes display a disruption in their cytoskeleton and the microtubule-associated protein, tau is abnormally phosphorylated. Control experiments were performed with the p105 antibody from an independent source, and with an antibody against Ki-67. RESULTS: The antibody against p105 (but not against Ki-67) immunolabeled the tangles, neuropil threads, and a subpopulation of fusiform neurites in the plaque. The p105 immunoreactivity was colocalized with tau and the numbers of tangles immunostained with p105 antibody correlated with the number of tangles immunolabeled with an antibody against tau. Western blot analysis of human brain homogenates showed that the antibody against p105 immunostained a major band at an estimated molecular weight of 105 kilodaltons and a secondary band at 84 kilodaltons. In AD frontal cortex, there was a significant increase in the 84 kilodalton band in the particulate, but not in the cytosolic fraction. CONCLUSIONS: The characterization of the patterns of expression of this class of nuclear antigen in the brain might provide important clues as to the mechanisms of plasticity and abnormal growth under normal conditions, as well as in AD. PMID- 8246449 TI - Pituitary specific transcription factor messenger ribonucleic expression in adenomatous and nontumorous human pituitary tissues. AB - BACKGROUND: The pituitary-specific transcription factor, Pit-1/GHF-1 (Pit-1) is a member of the POU family of homeobox genes. This transacting factor has a role in the development and expression of selected anterior pituitary cells including prolactin, growth hormone (GH), and thyrotropin, cell types in rodent pituitary. The expression of Pit-1 mRNA in human pituitaries has not been previously analyzed and the role of specific secretagogues in the regulation of Pit-1 transcripts in human pituitary adenomas has not been examined. Analysis of Pit-1 expression in normal and neoplastic human pituitaries may provide insight into the specificity of this transcription factor for pituitary cell types and the role of Pit-1 in human pituitary function. EXPERIMENTAL DESIGN: In situ and Northern hybridization analyses were used to study the expression of Pit-1 mRNA in surgically removed human pituitary adenomas and nontumorous pituitaries obtained within 6 hours postmortem. A cultured GH surgically removed adenoma was used to analyze the regulation of GH and Pit-1 in vitro. RESULTS: In situ hybridization localized Pit-1 mRNA in all pituitary adenomas, although the mRNA levels in GH and prolactin adenomas were significantly higher than those in ACTH, null cell and gonadotroph tumors. Northern hybridization analysis revealed a 2.4 and a 1.2 kb mRNA transcript for Pit-1. Although GH mRNA transcript was markedly changed by dexamethasone and phorbol 12-myristate 13-acetate in vitro, Pit-1 mRNA transcripts were not changed significantly by these secretagogues. CONCLUSIONS: Pit-1 mRNA transcripts are expressed mainly in prolactin and GH pituitary adenomas, but are also found in other adenomas and in the nontumorous adult human pituitary. Pit-1 mRNA transcript levels are not readily modulated in cultured GH adenoma cells by various secretagogues in vitro compared with the marked modulation of GH mRNA transcripts by these same secretagogues. PMID- 8246450 TI - Double infection with human papillomavirus 1 and human papillomavirus 63 in single cells of a lesion displaying only an human papillomavirus 63-induced cytopathogenic effect. AB - BACKGROUND: Specific histologic features are associated with the presence of certain human papillomaviruses (HPV) in verrucous lesions of the skin. HPV 1 is known to be associated with eosinophilic keratohyalin-like intracytoplasmic inclusion bodies, granular in appearance, whereas the recently described HPV 63 seems to induce inclusion bodies displaying a filamentous structure. Although an infection by two distinct HPV types within a single cell has not been previously reported, we have recently treated a plantar punctate keratotic wart in which only the histologic features seen in an HPV 63-induced lesion were seen, in spite of the detection of double infection of both HPV 1 and HPV 63 DNAs, thus posing the question as to the role of the HPV 1 present in the lesion. EXPERIMENTAL DESIGN: To localize the genomes of these two viruses within the lesion, double fluorescence in situ hybridization was performed using biotin-labeled HPV 63 DNA and digoxigenin-labeled HPV 1 DNA. The distribution of the genomes of these two viruses was examined in comparison with the histologic features in the lesion. The copy numbers of HPV 63 and HPV 1 were compared on agarose gel electrophoresis, stained with ethidium-bromide. RESULTS: We demonstrated the presence of the DNA of both HPV 1 and HPV 63 within the same nucleus of a plantar wart. In spite of the double infection of these two viruses in a cell and more or less equal copy numbers of the viruses, the cytopathogenic effect of only HPV 63 was seen in the cells. CONCLUSIONS: This is the first report of double infection of two different types of HPV within the same nucleus. Our findings suggest the possible interference that two HPV types could have on each other in inducing the HPV type specific cytopathogenic effects. PMID- 8246451 TI - Myofibroblast-derived smooth muscle cells during remodelling of rabbit urinary bladder wall induced by partial outflow obstruction. AB - BACKGROUND: Fibrosis of serosa, along with smooth muscle (SM) cell hypertrophy, has been shown to occur in the rabbit bladder after partial outflow obstruction. Identification of cells involved in the serosal thickening can be of primary interest to elucidate the functional changes that this organ undergoes. EXPERIMENTAL DESIGN: Cytoskeletal protein composition of cells present in the thickened serosa at different times from the onset of obstruction (7, 15, 30 and 60 days) was evaluated. This was accomplished by means of a panel of monoclonal antibodies specific for a number of differentiation markers of mesenchymal cells (vimentin, desmin, alpha-actin of SM type, nonmuscle (NM) and SM myosins), and by immunocytochemical and immunochemical techniques. RESULTS: The immunocytochemical study revealed that cells in serosal thickening follow a two-step maturation process from pre-existing vimentin-positive cells. In the first time period (7 to 15 days of obstruction), these cells predominantly achieved an immunophenotype corresponding to that of a specific myofibroblast subtype (i.e., containing vimentin, NM myosin, and SM alpha-actin). After 30 days from the onset of obstruction, the cytoskeletal protein content of serosal cells, as also revealed by Western blotting experiments, shifted towards that of fetal-type SM cells (i.e., presence of vimentin, NM myosin, SM alpha-actin, and SM myosin isoforms). Distribution of vimentin, desmin, SM alpha-actin, and SM myosin in tissue culture as well as the ultrastructure in vivo very closely resembled that of SM cells. Bromodeoxyuridine incorporation studies indicated that cells accumulated in the serosa of obstructed bladders did not derive, at least initially, from SM cells of the detrusor muscle. CONCLUSIONS: These findings are consistent with the existence of a differentiation process in which resident mesenchymal cells of bladder serosa may transform to myofibroblasts and, subsequently, in fetal-type SM cells during experimental outflow obstruction. PMID- 8246452 TI - A selective culture system for generating terminal deoxynucleotidyl transferase positive lymphoid cells in vitro. III. Structure of the bone marrow microenvironment for early lymphopoiesis. AB - BACKGROUND: We have previously demonstrated the feasibility of generating terminal deoxynucleotidyl transferase-positive (TdT+) lymphoid precursor cells in vitro in the nonadherent compartment of a long-term xenogeneic culture system in which rat bone marrow (BM) cells are seeded onto established mouse BM adherent cell layers. We have also noted that the appearance of TdT+ cells in these cultures is preceded by the formation of clusters of lymphoblasts in close association with the mouse BM adherent cell layer. Inasmuch as the selective generation of such primitive lymphoid cells is not ordinarily observed in homogeneic (i.e., mouse: mouse, rat:rat) BM cultures, the nature of the microenvironment for the generation of committed lymphoid stem/progenitor cells has not yet been detailed. Consequently, the aim of this study was to define the cellular components in the adherent compartment of our xenogeneic culture system that are associated with the earliest stages of lymphopoiesis in vitro. EXPERIMENTAL DESIGN: The nature of the interactions between rat BM lymphoid precursor cells and mouse BM adherent microenvironmental cells was investigated by a combination of immunophenotyping and scanning and transmission electron microscopy of primary cultures. The kinetics of formation and composition of lymphoid clusters were also determined morphologically and phenotypically. Results were compared with those of other investigators who have studied lymphopoiesis in intact BM or in homogeneic cultures of pre-B cells. RESULTS: Two distinct microenvironmental regions are represented within the mouse BM adherent cell layer: (a) paucilayer (PL) regions, composed of two or three horizontally oriented layers of alkaline phosphatase-positive mouse stromal cells; and (b) multilayer (ML) regions, containing 4 to 8 layers of such stromal cells. In both regions, proliferating rat lymphoid cells, expressing the HIS24 (B220) and/or HIS50 (heat stable antigen) early B-lineage antigens, are "sandwiched" between adjacent layers of stromal cells and enveloped by cytoplasmic processes from interdigitating mouse macrophages (pseudoemperipolesis). More than 95% of the lymphoid cells are of rat origin, whereas more than 95% of the nonlymphoid cells are of mouse origin. Large clusters, containing up to 1,000 lymphoid cells, preferentially develop in the ML regions and are comprised primarily of TdT+ cells. Small clusters containing 5 to 50 lymphoid cells, preferentially develop in the PL regions and are comprised primarily of TdT- cells, that can generate TdT+ cells upon transfer onto fresh adherent cells layers. Formation of individual small clusters, which outnumber large clusters by approximately 10 fold, is initiated by as few as 25 unfractionated rat BM cells. This process is not preceded by a lag period, and is linear with respect to time and cell dose. Formation of large clusters requires approximately 30 times more input cells, and is linear with respect to time after a lag of 5 days. CONCLUSIONS: The number of small lymphoid clusters formed in vitro closely approximates the frequency of lymphoid stem/progenitor cells in the BM inoculum (3 to 5%). This suggests that, under ideal conditions, individual clusters are clonally derived and the seeding efficiency of the culture system approaches 100%. The results further suggest that large clusters are formed by the coalescence of numerous small clusters within ML regions of the adherent cell layer; and that the formation of ML regions may be supported by an underlying monolayer of macrophages. A novel aspect of this system appears to be the frequency of pseudoemperipolesis, rather than phagocytosis, of primitive lymphoid cells by macrophages, that has also been noted in vivo. Moreover, the ML regions themselves bear a close resemblance to the recently described pro-B cell-enriched, multicellular aggregate fraction of freshly harvested mouse BM. Hence, this system appears to structurally recreate in vitro the PMID- 8246453 TI - Simple assay system for detecting human T cell leukemia virus type I-binding cells and its application in titrating binding inhibitory antibodies. AB - BACKGROUND: A simple and rapid assay system for the detection of human T cell leukemia virus type I (HTLV-I) binding cells was developed to assess the virus specific receptor and titrate the antibodies to block the virus binding. EXPERIMENTAL DESIGN: Cells (5 x 10(5)) were incubated with 100 microliters of the concentrated HTLV-I at 37 degrees C for 1 hour. After washing, the cells were reacted with anti-HTLV-I envelope monoclonal antibody (rat) for 30 minutes on ice and then stained with fluorescein-isothiocyanate-conjugated anti-rat immunoglobulin. The stained samples were analyzed on FACScan. Antibody-titration of the virus-binding inhibition was carried out by pretreatment of the virus with serially diluted sera. RESULTS: The specificity of the virus-binding was shown by dose-response relationship, kinetics of the binding, and temperature dependency. HTLV-I was absorbed onto a wide range of human cell lines and peripheral blood lymphocytes at various levels. Antibodies to inhibit the virus-binding were also quantitatively detected in sera from HTLV-I infected individuals, including asymptomatic carriers and patients with adult T cell leukemia or HTLV-I associated myelopathy, but not from healthy seronegative controls. CONCLUSIONS: This assay system would be useful in screening the virus-specific receptor and the neutralizing antibodies to HTLV-I. Thus, the assay could be applied to further studies on HTLV-I-related diseases. PMID- 8246454 TI - Community psychiatry in South Carolina: current status and future directions. PMID- 8246455 TI - Trends in public psychiatric inpatient care in a general hospital. PMID- 8246456 TI - Mobile crisis: comprehensive emergency psychiatry for the future. PMID- 8246457 TI - Treatment and research issues: bipolar affective disorder and substance abuse. PMID- 8246458 TI - Building a community-based system of mental health services. PMID- 8246459 TI - How can schools help children from homeless families? AB - Families represent the fastest growing subgroup among the homeless in America. Most homeless families are headed by women who lack high school diplomas, have poor work skills, little job training, and are long-term users of Aid to Families with Dependent Children (AFDC). The average homeless family is comprised of two to three children, most of whom are younger than age five. These children suffer from a number of acute and chronic health problems. Once in school, these children often experience developmental and academic delays which increase the potential for school dropout. School districts must address the health and academic challenges these children bring to school. Measures include forming a community-based task force, working with homeless shelters in addressing needs of these families, increasing awareness of faculty and staff, and developing specific classroom/school environment strategies to encourage these children to succeed. Opportunities for involvement by teachers, counselors, and school nurses are noted, and seven strategies to help the homeless child in school are offered. PMID- 8246460 TI - Evaluation of access to care and medical and behavioral outcomes in a school based intervention program for attention-deficit hyperactivity disorder. AB - The Project for Attention-Related Disorders (PARD), instituted by the San Diego (Calif.) School District. represents a unique approach to identification, referral, and management of children with Attention-Deficit Hyperactivity Disorder. As a school-based program, PARD coordinates medical, psychosocial, behavioral, and educational interventions for each child, and provides resources for school personnel caring for children with ADHD. Three years after instituting PARD, about 60% to 70% of the children showed improvement after enrollment, evaluation, and treatment. Parental involvement in and acceptance of intervention for their children represented a significant follow-up problem. Both medical and nonmedical interventions such as counseling, special programs, and behavior modification helped improve the educational life of these children. PMID- 8246461 TI - A cardiovascular health education program for rural schools. AB - Public understanding of cardiovascular disease (CVD) risk factors and primary prevention has increased, due in part to community prevention efforts. However, many segments of society are difficult to reach. Such groups still need public education to acquire the knowledge that can lead to behavior change. Community intervention programs in rural areas face the challenge of disseminating health information to widely scattered populations isolated by difficult terrain and weather, and restricted by the sparsity of channels for mass communication. School health promotion programs, because of the special role schools play in rural communities, can help reach rural populations. During a five-year period, the Otsego-Schoharie Healthy Heart Program, a state-funded community intervention program, provided presentations to 18% of the combined total population of two rural counties through its school-based component. It also helped promote other program initiatives by establishing linkages in the community. Schools provide an effective channel for health promotion efforts to reach rural populations. PMID- 8246462 TI - Cigarette smoking as a predictor of alcohol and other drug use by children and adolescents: evidence of the "gateway drug effect". AB - Data from a statewide survey, conducted by the Indiana Prevention Resource Center, of 20,629 Indiana students in grades 5-12 were analyzed to determine the extent to which cigarette smoking predicted use of alcohol and other drugs and acted as a so-called "gateway drug." A three-stage purposive/quota cluster sampling strategy yielded a representative sample of Indiana students, stratified by grade. Cross-tabulated data revealed a strong, dose-dependent relationship between smoking behavior and binge drinking, as well as use of alcohol and illicit drugs. Daily pack-a-day smokers were three times more likely to drink alcohol, seven times more likely to use smokeless tobacco, and 10-30 times more likely to use illicit drugs than nonsmokers. A stepwise multiple regression analyzed the role that the student's perceptions of the risk of using drugs and of peer approval/disapproval of the student's drug use, gender, grade in school, and ethnic background played in predicting alcohol and other drug use. PMID- 8246463 TI - Educating school nurses to care for HIV-infected children in school. AB - Effects of an educational program on school nurses' knowledge and attitudes about human immunodeficiency virus (HIV) infection in children and changes in practice was investigated. Participants (N = 205) attended a one-day workshop held at 11 sites in New Jersey. Content was clinically focused and derived from a needs assessment of a sample of school nurses throughout New Jersey. An adult education format was used, with interactive and affective teaching strategies modeled on the AIDS Training Program of the California Nurses' Association. Subjects were pretested and posttested for knowledge and attitudes about HIV in children. An identical posttest mailed three months later included a self-report of change in practice since attending the workshop. Knowledge and attitude scores were significantly higher at program completion (p < .0001). Three months after the program, knowledge scores had fallen but not to pretest levels. No significant difference existed between attitude scores after the program and three months later. On self-report of change in practice, respondents indicated they institute or improved universal precautions, and were more comfortable in teaching children, staff, and their communities about HIV. PMID- 8246464 TI - Schools and health: a natural partnership. PMID- 8246465 TI - A method to increase informed consent in school health research. PMID- 8246466 TI - The effects of early bilateral otitis media with effusion on language ability: a prospective cohort study. AB - The current study, which is a follow-up on the epidemiological Nijmegen Otitis Media study, examines the relationship between early otitis media with effusion (OME) and later language ability in a group of children with systematically documented bilateral OME. In the Nijmegen Otitis Media study, children were screened using tympanometry at regular intervals of 3 months, between their second and fourth birthdays. At age 7, three groups of children participated in language testing: 82 OME-free children, 151 children with early bilateral OME, and 37 children treated with ventilation tubes at preschool age. A history of OME, even up to nine instances, did not have negative consequences for language performance at age 7. Intermittent, as opposed to more continuous, OME was not found to affect language ability negatively. The suggested benefit of treatment with ventilation tubes was not found. PMID- 8246467 TI - The influence of prosodic and gestural cues on novel word acquisition by children with specific language impairment. AB - The purpose of this study was to investigate the effects of prosodic and gestural cues on children's lexical learning. Acquisition of novel words was examined under linguistic input conditions that varied in terms of rate of speech, stress, and use of supplemental visual cues (i.e., gestures). Sixteen kindergarten children served as subjects in this study, including 8 children with normal language (NL) and 8 children with specific language impairment (SLI). A repeated measures design was used such that all subjects in both groups participated in each of the three experimental conditions (the Rate, Stress, and Visual Condition). Results indicated that acquisition of novel words by the groups with NL and SLI was significantly affected by alterations in speaking rate and by the use of gestures accompanying spoken language. There were no statistically significant effects for the stress manipulations, although subjects with SLI tended to correctly produce novel words that had received emphatic stress during training more often than words presented with neutral stress. Implications of these findings are discussed with respect to the importance of considering how the manner of presentation of the linguistic signal influences the processing and acquisition of language. PMID- 8246468 TI - The performance of children with language-learning disabilities on two types of cognitive tasks. AB - The performance of children with language-learning disabilities on two types of cognitive tasks was compared to that of two groups without language-learning disabilities, one matched for mental age and the other for language (vocabulary) age. The first task, Concept Formation, measured one's ability to identify and state the rule for a concept about a set of colored geometric figures when shown instances and noninstances of the concept. The second task, Analysis Synthesis, required the identification and use of a solution key to solve an incomplete logic puzzle. Both tasks involved increases in complexity as additional items were administered. Results indicate that the children with language-learning disabilities performed at a level commensurate with the language-matched group on the Concept Formation task, and at a level commensurate with the mental-age matched group on the Analysis-Synthesis task. The implications of these findings for the nature of language-learning disabilities are discussed. PMID- 8246469 TI - Comparison of two methods for promoting productive vocabulary in late talkers. AB - This investigation compared the effectiveness of two language treatment methods, modeling versus modeling plus evoked production, in promoting productive vocabulary in three toddlers identified as late talkers. A single-subject alternating treatments design was employed in this study in which different sets of words were taught under the two treatment methods during group and individual instruction. Some evidence of differential patterns of response to the treatment types was found for two subjects, but the subjects differed as to which particular treatment method was associated with better performance. Neither treatment method was effective for the third subject. Dynamic assessment measures were only marginally useful in predicting overall lexical learning potential in these subjects, and did not predict response to the two teaching methods. Implications of these results are discussed with regard to the role of language intervention for late talkers. PMID- 8246470 TI - Constituent syllable effects in a nonsense-word repetition task. AB - Multisyllabic nonsense-word repetition tasks have been used to provide evidence on the phonological processing operations of children with language impairment, independent of their lexical knowledge (Gathercole & Baddeley, 1990; Kamhi, Catts, Mauer, Apel, & Gentry, 1988). However, recent evidence (Gathercole, Willis, Emslie, & Baddeley, 1991) and speculation (Snowling, Chiat, & Hulme, 1991) suggest that the nonsense words employed in such tasks may not be equally "nonsensical." The present investigation directly tested the effect on repetition performance of one previously uncontrolled characteristic of multisyllabic nonsense words: the lexical status (word or nonword) of their stressed syllables. Normally achieving school-age boys repeated nonsense words with lexical stressed syllables significantly more accurately than nonsense words with nonlexical stressed syllables. These results suggest the need to control, at a minimum, the lexical status of constituent syllables in constructing nonsense-word stimuli. PMID- 8246471 TI - History of middle ear involvement and speech/language development in late talkers. AB - Late-talking and normally speaking toddlers with and without histories of middle ear involvement were followed for 2 years to assess speech and expressive language outcomes. Results revealed no differences in expressive language outcome that could be attributed to history of middle ear involvement in either group. There did seem to be differences in outcome on measures of articulation that were associated with history of middle ear involvement. The implications of these findings for treatment of otitis media and for referral of late-talking toddlers for speech and language services are discussed. PMID- 8246472 TI - The efficacy of using the IBM Speech Viewer Vowel Accuracy Module to treat young children with hearing impairment. AB - The efficacy of the IBM SpeechViewer's Vowel Accuracy Module for the treatment of vowel productions was evaluated in six preschool children with hearing-impairment over a 4-month period. A single-subject design was used, and the vowels /a/, /i/ and /u/ were treated. Untreated sounds also were probed to monitor for carryover and developmental effects. One of the children was dismissed from the study because of noncompliance. Of the remaining five children, four exhibited a treatment effect for /u/, two for /a/, and one for /i/. Four of the children demonstrated some generalization. Developmental effects, as represented by change in /s/-cluster production, were not documented. Although treatment effects were observed, difficulties with the Vowel Accuracy Module were also observed. These included inaccuracies in the feedback on low-intensity, hypernasal, and high pitched utterances; inability to sustain the attention of preschoolers over multiple sessions; lack of instructional feedback; and nonlinearity in the criterion-adjustment control. PMID- 8246473 TI - Intelligibility of selected passages from the Speech Intelligibility Rating (SIR) test. AB - Two experiments were conducted to examine the intelligibility of 12 of the 72 passages of connected discourse prepared by Cox and McDaniel (1984, 1989) in the development of the Speech Intelligibility Rating (SIR) test. Intelligibility was assessed with a method-of-adjustment procedure in the presence of two maskers. One was a multi-talker babble with a variable S/N ratio environment that yields intelligibility scores that are potentially level-dependent because of the almost inevitable difference in speech intensity from passage to passage. The second was a signal-correlated noise with a constant S/N ratio environment that provides scores that are essentially level-independent. Two homogeneous subsets of nine passages each were identified that yield equivalent intelligibility scores. The outcome underscores the value of incorporating a signal-correlated noise masker that yields scores that are relatively unaffected by small differences in signal level among passages. PMID- 8246474 TI - Auditory stroop effects in children with hearing impairment. AB - The accurate perception of speech involves the processing of multidimensional information. The aim of this study was to determine the influence of the semantic dimension on the processing of the auditory dimension of speech by children with hearing impairment. The processing interactions characterizing the semantic and auditory dimensions were assessed with a pediatric auditory Stroop task. The subjects, 20 children with hearing impairment and 60 children with normal hearing, were instructed to attend selectively to the voice-gender of speech targets while ignoring the semantic content. The type of target was manipulated to represent conflicting, neutral, and congruent relations between dimensions (e.g., the male voice saying "Mommy," "ice cream," or "Daddy" respectively). The normal-hearing listeners could not ignore the irrelevant semantic content. Instead, reaction times were slower to the conflict targets (Stroop interference) and faster to the congruent targets (Stroop congruency). The subjects with hearing impairment showed prominent Stroop congruency, but minimal Stroop interference. Reduced Stroop interference was not associated with chronological age, a speed-accuracy tradeoff, a non-neutral baseline, or relatively poorer discriminability of the word input. The present results suggest that the voice gender and semantic dimensions of speech were not processed independently by these children, either those with or those without hearing loss. However, the to be-ignored semantic dimension exerted a less consistent influence on the processing of the voice-gender dimension in the presence of childhood hearing loss. The overall pattern of results suggests that speech processing by children with hearing impairment is carried out in a less stimulus-bound manner. PMID- 8246475 TI - The influence of noise on the measured amplitudes of distortion-product otoacoustic emissions. AB - This paper describes the influence of noise on the measured amplitudes of tonal signals, as determined using narrowband spectral analysis, that is, the technique typically used to measure distortion-product otoacoustic emissions (DPOAEs). At small signal-to-noise ratios, background noise results in overestimation of DPOAE amplitude and, thus, substantially influences the measured properties of the low level portions of DPOAE-growth functions, in particular the apparent slope of the functions in this region. It is shown that, because of the influence of noise, the algorithm for the objective estimation of detection thresholds of DPOAEs, and of the slopes of DPOAE-growth functions, described by Nelson and Kimberley (1992), will tend to underestimate these values. This systematic underestimation is presumably the reason why many of the DPOAE-detection thresholds and growth slopes presented in that study were considerably lower than those reported in previous studies using similar measuring equipment and paradigms but different detection-threshold and growth-slope estimation techniques. In the present paper, a simple equation allowing an estimated correction for the effects of noise on measured DPOAE amplitudes is presented. Finally, an alternative strategy for the estimation of DPOAE thresholds, one that is less prone to the influence of noise, is suggested. PMID- 8246476 TI - Narrative discourse in children with language disorders and children with normal language: a critical review of the literature. AB - This review and analysis of the literature on narrative discourse in children places particular emphasis on children with language disorder. The review (a) describes theoretical perspectives on narrative use, (b) surveys researchers' rationales for the investigation of narrative ability, (c) discusses methodological issues relevant to narrative research, and (d) concludes with a discussion regarding future research. Specific topics contained within these discussions include contributions from allied disciplines, the pragmatic nature of narrative use, narrative ability as an index of language development, methodological issues in research design, and clinical implications of future research. PMID- 8246477 TI - F2 transitions during sound/syllable repetitions of children who stutter and predictions of stuttering chronicity. AB - The purpose of the present study was to examine the relationships between second formant (F2) transitions during the sound/syllable repetitions (SSRs) of young children who stutter and their predicted chronicity of stuttering. Subjects were 13 youngsters who stutter, who were divided into two groups based on their predicted chronicity of stuttering as measured by the Stuttering Prediction Instrument (SPI; Riley, 1984): a high-risk group, consisting of 7 boys (mean age = 50.6 months), and a low-risk group, consisting of 5 boys and 1 girl (mean age = 48.5 months). Each child was audio/videotape-recorded during a 30-minute conversational interaction with his or her mother. Ten SSRs per child were acoustically analyzed to identify differences in F2 transitions between the repeated (stuttered) and fluent (nonstuttered) portions of the words. Present findings are consistent with those of Stromsta (1965, 1986), who reported that children who stutter produce F2 transitions during stuttering that are nonmeasurable or missing or that differ in direction of movement from fluent transitions. However, there were no significant between-group differences in the frequency of occurrence of these "abnormal" F2 transitions, findings that are apparently inconsistent with Stromsta's results. The remaining measurable F2 transitions showed no significant between-group differences in the mean differences between stuttered and fluent F2 transitions for onset and offset frequencies, transition extents, and transition rates. Within both groups, significant positive correlations were found between stuttered and fluent F2 transitions for all acoustic measures except for transition durations, which were not significantly correlated for either high-risk or low-risk subjects. Within the low-risk group, stuttered F2 transitions were typically shorter than fluent transitions. Findings were taken to suggest that some elements of sound or segment prolongation may be present within the SSRs of children who stutter and who are considered to be at high risk for continuing to stutter, indicating that further study of selected aspects of F2 transitions during stuttering may provide useful clinical information for predicting the likelihood that a child will continue to stutter. PMID- 8246478 TI - Event-related brain potentials preceding speech and nonspeech oral movements of varying complexity. AB - Cortical preparation for movement is reflected in the readiness potential (RP) waveform preceding voluntary limb movements. In the case of oral movements, the RP may be affected by the complexity or linguistic nature of the tasks. In this experiment, EEG potentials before a nonspeech task (lip pursing), a speech-like task (lip rounding), and single word production were recorded from scalp electrodes placed at the cranial vertex (Cz) and over the left and right motor strips (C3' and C4'). Seven right-handed female subjects produced at least 70 repetitions of the three tasks, in each of five repeated sessions. EEG records were averaged with respect to EMG onset at the lip. The word task, as opposed to the other tasks, was associated with greater negative amplitude in the RP waveform at the vertex site. Differences between the waveforms recorded at the right- and left-hemisphere sites were insignificant. Although intersubject variability was high, individuals had relatively stable patterns of response across sessions. Results suggest that the RP recorded at the vertex site is sensitive to changes in task complexity. The RP did not reflect lateralized activity indicative of hemispheric dominance. PMID- 8246479 TI - Coexistence of stuttering and disordered phonology in young children. AB - The purpose of the present study was to assess differences in stuttering, phonological, and diadochokinetic behaviors in young children who exhibit both stuttering and disordered phonology and children who exhibit only one of the disorders. Subjects were 21 male children (aged 4 to 6 years), representing three groups of seven children each: (a) stuttering and normal phonological abilities (S+NP), (b) stuttering and disordered phonology (S+DP), and (c) normal fluency and disordered phonology (NF+DP). Stuttering behavior was assessed during a 30 minute conversational speech task; phonological behavior was assessed during a 162 item picture-naming task; and diadochokinetic abilities were assessed during bi- and multisyllable productions. Results indicated that the S+DP group produced significantly more sound prolongations and significantly fewer iterations per whole-word repetition than the S+NP group. However, there were no differences between the two groups in other stuttering indices. Moreover, no differences were noted between the S+DP and NF+DP groups in phonological behavior. Diadochokinetic rates did not differ among the three groups. The possibility of two types of stuttering, one occurring with and one without disordered phonology, is discussed. PMID- 8246480 TI - Speech and swallow function after tonsil/base of tongue resection with primary closure. AB - Speech and swallowing function was examined in 11 patients who underwent surgical resection of greater than 1 cm of tongue base, tonsil, and faucial arch with mandible resected on the side of the tumor and reconstruction by primary closure. Preoperatively and 1 and 3 months post-healing, high fidelity audio recordings were made of a 6- to 7-minute conversational speech sample, the sentence version of The Fisher Logemann Test of Articulation Competence was administered, and videofluoroscopic assessment of oropharyngeal swallow was conducted. All subjects exhibited changes in speech and swallowing function postoperatively, with little improvement during the study. Patients exhibited greatest difficulty on stop and fricative consonants and bolus propulsion. Comparison with patients who received anterior tongue and floor of mouth resections and distal flap reconstruction revealed consistently better speech performance by the tonsil/base of tongue patients, although the same phonemes were affected. Swallow function was equally affected in the two groups. Results are discussed in terms of locus of surgical resection, nature of reconstruction, and need for swallowing therapy. PMID- 8246481 TI - The need for replication. AB - Replication provides verification and disconfirmation functions for the scholarly fields. Relatively few replications were found in a survey of studies in JSHD and JSHR over a recent decade. Based on the probabilities of Type I and Type II errors, there are likely to be approximately 50 to 250 false findings in this literature. Because many studies had relatively small sample sizes, replication would be helpful for extending the generalization of their results. These data underscore the urgency for more replications in the field. PMID- 8246482 TI - Phonologic error distributions in the Iowa-Nebraska Articulation Norms Project: word-initial consonant clusters. AB - The errors on word-initial consonant clusters made by children in the Iowa Nebraska Articulation Norms Project (Smit, Hand, Freilinger, Bernthal, & Bird, 1990) were tabulated by age range and frequency. The error data show considerable support for Greenlee's (1974) stages in the acquisition of clusters: the youngest children show cluster reduction, somewhat older children show cluster preservation but with errors on one or more of the cluster elements, and the oldest children generally show correct production. These stages extended to three element clusters as well. Typical cluster reduction errors were (a) reduction to the obstruent in obstruent-plus-approximant clusters and (b) reduction to the second element in /s/-clusters. When clusters were preserved, but one member was in error, the error was typically the same as for the singleton consonant. Cluster errors are discussed in terms of theories of phonologic development, including open genetic programs and feature geometry. These data are expected to be useful in evaluation and treatment of disorders of phonology. PMID- 8246483 TI - The relationship between speech-language impairments and reading disabilities. AB - A group of children with speech-language impairments was identified in kindergarten and given a battery of speech-language tests and measures of phonological awareness and rapid automatized naming. Subjects were followed in first and second grades and administered tests of written word recognition and reading comprehension. The children with speech-language impairments were found to perform less well on reading tests than a nonimpaired comparison group. Subjects' performance on standardized measures of language ability in kindergarten was observed to be closely related to reading outcome, especially reading comprehension. Measures of phonological awareness and rapid automatized naming, on the other hand, were found to be the best predictors of written word recognition. The implications of these findings for the early identification and remediation of reading disabilities are discussed. PMID- 8246484 TI - The emergence of mature gestural patterns is not uniform: evidence from an acoustic study. AB - Previous studies investigating the organization of articulatory gestures present conflicting accounts of age-related differences in the execution of the articulatory gestures themselves and in the organization of those gestures. Several methodological differences may help to explain these contradictions: First, different studies have used different measures, all of which reflect vocal tract activity to varying extents; second, the articulatory gestures being analyzed differed across studies; third, the phonetic composition of syllables has varied; and finally, utterance length, and therefore complexity, has varied across studies. The purpose of this study was to investigate the possibility that the reason these methodological differences have led to contradictory results is because the emergence of mature gestural patterns in children's speech is not uniform. To accomplish this goal, detailed acoustic analyses were performed on schwa-stop-vowel utterances from adults and from children (3, 5, and 7 years of age). Temporal measures showed that some acoustic segments were longer in children's than in adults' samples, whereas others were similar in duration. Formant frequencies indicated that vocal-tract opening and closing achieve adult like patterns of movement by the age of 3 years, but children's tongue gestures are constrained by phonetic context more than those of adults until at least the age of 7 years. Taken together, these results suggest that the pace of development for learning to produce and to coordinate articulatory gestures is not uniform. Thus, the contradictions in findings among earlier studies may very well reflect differences in choices of measurement and utterances to be analyzed, both of which may lead to evaluations of different aspects of gestural patterning. PMID- 8246485 TI - Conversations with children who are language impaired: asking questions. AB - Samples of conversational language were elicited with a standardized interview protocol from 24 children, aged 2:6 to 7:8, half with specific language impairment (SLI), half with normally developing language (LN), matched for language level. Samples were analyzed to determine whether there were associations between adult questioning and children's use of ellipsis. For the SLI children, but not the LN children, increased proportions of questions were significantly correlated to increased proportions of ellipsis. This finding has implications for the use of MLU measures in clinical and research practice. PMID- 8246486 TI - Effects of initial element teaching in a story-telling context on Blissymbol acquisition and generalization. AB - Blissymbolics is a graphic symbol system used by individuals with little or no functional speech. The purpose of this study was to determine the effects of initial teaching of semantic elements on compound Blissymbol acquisition, retention, and generalization in a story-telling context. The subjects included 40 preschool children with normal cognitive abilities (mean chronological age = 45 months) who were assigned to one of two groups. Group I was taught elements before being taught compounds that contained these elements. Group II was taught elements before being taught compounds consisting of elements that were not taught previously. In addition, both groups received instruction in a second set of compounds that were taught directly, that is, without first being taught the elements. The teaching procedures and materials were socially validated by experts in Blissymbol instruction. Results indicate that the initial teaching of elements did not contribute to compound acquisition and retention, but did facilitate generalization to untrained compound Blissymbols. The results are discussed in terms of considerations for selecting an initial lexicon and Blissymbol teaching and research. PMID- 8246487 TI - Auditory lexical decisions in children and adults: an examination of response factors. AB - This study examined the influence of nonlexical response factors on the speed of auditory lexical decisions in children and adults. Two groups of children (6- and 7-year-olds, 8- and 9-year-olds) and adults participated in three tasks: a real word lexical decision task in which subjects were asked to say "yes" as quickly as possible to real words; a nonword lexical decision task in which subjects were asked to say "no" as quickly as possible to nonwords; and an auditory-vocal reaction time task in which subjects were asked to say "yes" or "no" to a tone. Response times on all tasks decreased with age. However, the age-related differences on the real-word lexical decision task disappeared when differences in auditory-vocal reaction times were taken into account. This result suggests that a large part of developmental differences in the speed of lexical processing may be due to nonlexical response factors. PMID- 8246488 TI - Kupffer cells can present alloantigen in vitro: an effect abrogated by gadolinium. AB - Portal venous (PV) injection of alloantigen can result in tolerance. We have previously reported that this effect is abrogated by pretreatment with gadolinium (Gd), an element known to inhibit Kupffer cell phagocytosis. To further elucidate the role of Kupffer cells (KC) in PV tolerance, the present study examined the ability of KC to present alloantigen in vitro to syngeneic responder lymphocytes after in vivo PV administration of alloantigen with or without pretreatment with Gd. Wistar Furth (WF) rats were pretreated 24 hr prior to KC harvest with a PV injection of 1 x 10(7) allogeneic Lewis (LEW) rat lymphocytes (KC-L) or saline (KC-N). Another group received Gd intravenously 24 h prior to the PV injection of LEW cells (KC-L + Gd). KC were isolated from the WF rats using collagenase digestion and Percoll (90% pure by morphologic criteria and by monoclonal antibodies KU-1 and ED-2). Responder WF lymphocytes were cocultured with media alone, or with KC as prepared above, for 72 hr. KC-L demonstrated significant stimulation of the WF responders (P = 0.02 vs KC-N). Gd abrogated this stimulation (P = 0.04, KC-L vs KC-L + Gd) but not by nonspecific inhibition of the cocultures (KC-N vs KC-L + Gd, P > 0.05, no difference). This study demonstrates that KC can effectively present PV alloantigen for the activation of naive syngeneic T lymphocytes, thereby further supporting the hypothesis that KC play an integral role in the induction of PV tolerance by presentation of antigen to responder T cells. PMID- 8246489 TI - Changes in hepatocyte NADH fluorescence during prolonged hypoxia. AB - Deprivation of oxygen reduces oxidative phosphorylation and rapidly causes an increase in cellular NADH which can be monitored by fluorimetry. Previous studies have established that increases in NADH fluorescence accurately reflect the impairment in oxidative phosphorylation which occurs during brief periods of acute hypoxia. However, the potential usefulness of fluorimetry for following longer, clinically relevant periods of ischemia has not been explored. We studied changes in NADH fluorescence in rat hepatocyte suspensions and in isolated-buffer perfused rat livers during hypoxia (pO2 < 50 mm Hg) for periods as long as 180 min. NADH fluorescence of hepatocyte suspensions consistently increased by about 15% after 13 to 15 min of hypoxia and coincided with a marked decrease in tissue ATP levels. Reoxygenation after 15 or 30 min of hypoxia resulted in recovery of ATP and NADH with minimal loss of viability, as measured by trypan blue exclusion. After 60 to 180 min hypoxia, the initial increase in NADH fluorescence was followed by a progressive, irreversible decline which correlated with decreased cell viability. Similar changes in NADH fluorescence were observed in isolated-perfused rat livers in which NADH fluorescence was monitored at the liver surface with a fiberoptic probe. Hypoxia for 30 min had no effect on NADH fluorescence, but hypoxia for longer periods caused a steady increase in fluorescence after 45-60 min. When hypoxia was prolonged (120 or 180 min), fluorescence peaked after 60-75 min and then declined progressively to levels below baseline. The greatest decrease in fluorescence was seen after 180 min of hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246490 TI - Determination of small bowel viability using quantitative myoelectric and color analysis. AB - Intraoperative assessment of viability in ischemic bowel remains a major unsolved problem in general surgery. Bowel viability was assessed in 31 dogs 24 hr after mesenteric arterial ligation in a 40-cm segment of ileum. The purpose of this study was to evaluate two new quantitative methods of viability assessment, electromyography (EMG) and reflection densitometry. These methods were compared with traditional methods of bowel viability assessment. EMG was measured using a specially designed probe which contains an electronic control unit for computer assisted data acquisition. A computer algorithm quantified the EMG. Reflection densitometry quantified bowel color in the red and blue spectrum. Presence of visible peristalsis, Doppler ultrasound, and a visual color grading system were also used in viability assessment. Each parameter was measured at 2-cm intervals along the ischemic segment. Resection and anastomosis of ischemic bowel were then performed. There were nine deaths from anastomotic leak, all resulting from further bowel necrosis. EMG was the only viability assessment parameter that correlated with survival. EMG at the resection margin in survivors was 29 +/- 3% vs 23 +/- 4% at the resection site of nonsurvivors (P < or = 0.047 by unpaired Students t test). These results suggest that quantitative EMG measurements may be useful in assessment of viability in ischemic bowel. PMID- 8246491 TI - Enhanced cardiac preservation with oxygenated University of Wisconsin solution. AB - The purpose of this study was to determine if oxygen delivery to rabbit cardiac allografts arrested and stored in University of Wisconsin solution (UWS) at 4 degrees C would affect preservation. Nineteen isolated rabbit hearts were rapidly excised and perfused at 80 mm Hg on an isovolumic modified Langendorff apparatus. A micromanometer was placed within a balloon and inserted into the left ventricle through the mitral valve orifice. Digitized pressure waveforms were collected at 11 known balloon volumes from 0.8 to 1.2 ml. Baseline data were obtained for all hearts while perfused with Krebs-Henseleit solution equilibrated with 95% O2:5% CO2 at 37 degrees C. All hearts were arrested with 30 ml of UWS (290 mOsm). The control group (N = 10) was stored in UWS at 4 degrees C for 8 hr, and the experimental group (N = 9) was perfused with oxygenated UWS (O2 content = 5.6 ml O2) at 4 degrees C for 8 hr at a pressure of 60 mm Hg (5-10 ml/min). Both groups were then reperfused with Krebs-Henseleit buffer at 80 mm Hg for 15 min at for postpreservation data acquisition. Left ventricular developed pressures over a physiologic range (pressure-volume area) and maximum positive and negative dP/dt were calculated. Recovery of left ventricular parameters as a percentage of the baseline values was determined. Mean pressure-volume area recovery in the nonperfused group was 40 +/- 7.9% versus the perfused group (71 +/- 7.0%, P = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246492 TI - Intestinal ischemia in the newborn: the role of intestinal maturation. AB - Premature infants are susceptible to intestinal ischemia during the newborn period when their intestinal tracts are functionally and structurally immature. Studies have shown that exogenous glucocorticoids hasten intestinal maturation. We investigated the effects of hydrocortisone on platelet activating factor (PAF) induced intestinal ischemia in the neonatal rat. On Postnatal Days 7-11, Sprague Dawley rats were given intraperitoneal (ip) injections of either saline (SAL) or hydrocortisone (HC; 50 mg/kg total). On Day 12, rats were injected with either PAF (2 micrograms/kg) or an equal volume of saline. After 2 hr the rats were sacrificed and sections were taken for histology. The remaining intestine was analyzed for maltase, lactase, myeloperoxidase (MPO), and xanthine oxidase (XO). Experimental groups were as follows: SAL (N = 8), received saline only; SAL+PAF (N = 8), received saline plus PAF; HC (N = 3), received hydrocortisone+saline; and HC+PAF (N = 5), received hydrocortisone plus PAF. XO was significantly decreased (P < 0.001) in the hydrocortisone-treated groups (HC + SAL = 16.36 +/- 18.42 units/g protein, HC + PAF = 17.33 +/- 9.06 units/g protein) vs the controls (SAL only = 108.90 +/- 20.24 units g/protein, SAL + PAF = 145.77 21.28 units/g protein). MPO was not significantly elevated in SAL + PAF (4.60 +/- 0.95 units/g protein) vs HC + PAF (2.18 +/- 0.80 units/g protein) in this study. Maltase was significantly elevated (P < 0.001) in the HC + PAF (241.46 +/- 40.6 mole/min/g protein) and HC + SAL (152.78 +/- 16.35 mole/min/g protein) vs saline only (28.35 +/- 5.77 mole/min/g protein and SAL + PAF (37.29 +/- 8.70 mole/min/g protein. Animals (7/8) in the SAL + PAF group developed ischemia by inspection and histologic exam.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246493 TI - Energy metabolism of the heart and the liver in brain-dead dogs as assessed by 31P NMR spectroscopy. AB - The energy metabolism of the heart and the liver was assessed in brain-dead dogs by 31P nuclear magnetic resonance spectroscopy. The mean blood pressure and pulse rate changed respectively from 136.1 +/- 9.3 mm Hg (mean +/- SEM) and 126.7 +/- 4.4/min in the control period, to 212.9 +/- 15.8 mm Hg and 146.7 +/- 13.3/min during Cushing phenomenon (CU period), and to 60.0 +/- 6.4 mm Hg and 60.0 +/- 2.9/min after completion of brain death (BD period). The ratio of creatine phosphate to inorganic phosphate (PCr/Pi) of the heart decreased from 5.00 +/- 1.08 to 3.24 +/- 0.80 in the CU period and increased to the levels of the control values in the early BD period. The intracellular pH of the heart decreased from 7.20 +/- 0.07 to 6.91 +/- 0.02 in the CU period and increased to 7.12 +/- 0.01 in the BD period. The positive relationship between the Pi/PCr and the rate-pressure product (BP x PR) indicates that the regulation of the oxidative metabolism by free ADP was well maintained except at some points in the CU period (Pi/PCr = 8.77 x 10(-6) BP x PR + 0.164, r = 0.704). This suggested that the remarkable hypotension and bradycardia in the BD period is not associated with cardiac energy failure. Although the energy states of both the heart and the liver in brain-dead dogs were adequately maintained in the BD period, the changes were different.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246494 TI - Reperfusion of duodenum and pancreas following 24 hr of cold storage in Perfadex or UW solution. AB - We have studied the immediate reperfusion and the graft outcome of the duodenum and pancreas separately in a porcine whole-organ pancreaticoduodenal allograft transplantation model using 24 hr of cold storage. Of 19 transplantations, 12 grafts were perfused and stored in Perfadex and 7 in University of Wisconsin (UW) solution. The organ weights before and after storage were determined in 6 grafts stored in Perfadex and 6 grafts stored in UW solution in order to quantify the degree of dehydration of the graft during storage. In 6 of the grafts perfused with Perfadex, a hyperosmotic salt dextran-containing solution (HSD) was perfused into the graft aorta at reperfusion. Duodenal reperfusion and reperfusion injury were studied by measuring mucosal pH (pHi) and by microscopic examination, respectively. Pancreatic reperfusion was studied by scoring the macroscopic appearance and microscopic examination. Daily blood glucose was used to determine the endocrine function and a secretin-cholecystokinin stimulation test was performed on the second postoperative day to evaluate the exocrine function. All duodenal grafts except 1 in the UW group showed a sufficient reperfusion according to pHi and all had microscopic changes suggesting a moderate reperfusion injury. The macroscopic appearance of pancreas was significantly worse in the UW group than in the two Perfadex groups. Only 1 of 7 grafts in the UW group showed signs of endocrine function on the first postoperative day compared with 11 of 12 in the Perfadex groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246495 TI - The effect of cyclosporine A dissolved in chremofore or in ethanol and of cortisone on the arterial release of prostacyclin. AB - Cyclosporine A has been suggested to increase thromboembolic complications after renal transplantation. Therefore, the effect of cyclosporine A (at the clinically used dose of 10 mg/kg) dissolved in either chremofore or ethanol on rabbit vascular prostacyclin release was investigated in an ex vivo perfusion system. The animals received the drugs intravenously either as a single injection the day before operation or daily for 1 month prior to operation. Rabbits given cyclosporine A dissolved in chremofore released less prostacyclin than controls, both after a single injection and after 1 month of daily injections. The vehicle chremofore also gave a significantly lower release of prostacyclin than the control. The response to arachidonic acid with increased release was equal in all groups. Cyclosporine A dissolved in ethanol did not alter the initial release, and ethanol alone did not influence the prostacyclin release. Cortisone depressed the vascular prostacyclin release after daily injections for 1 month, but did not after only one injection. Cyclosporine A dissolved in chremofore and cortisone given in combination did not result in an additive reduction. These findings indicate that the intravenous administration of cyclosporine A dissolved in chremofore, but not that dissolved in ethanol, as well as cortisone, might decrease the vascular defense against thrombus formation. The action of these substances is higher up in the arachidonic acid cascade than the cyclooxygenase level. PMID- 8246496 TI - Despite maintenance of systemic and regional perfusion, endotoxemia following complement depletion produces hepatocellular dysfunction. AB - Experimental endotoxemia causes hypotension and a reduction in regional blood flow, including hepatic blood flow. Complement depletion prior to endotoxemia is known to attenuate these perfusion deficits. We depleted complement with cobra venom factor prior to the administration of Escherichia coli lipopolysaccharide in Sprague-Dawley rats and studied the effects of this treatment on systemic hemodynamics, regional hepatic perfusion, and hepatocellular integrity. Complement-depleted endotoxemic rats were compared with untreated rats, rats with complement depletion alone, and rats with endotoxemia alone. Systemic hemodynamics (cardiac index, mean arterial pressure), regional hepatic perfusion (effective hepatic blood flow), and hepatocellular integrity (adenosine triphosphate [ATP], lipid peroxidation) were determined 4-6 hr after the onset of endotoxemia. The endotoxemic animals exhibited a significant decrease in systemic hemodynamic performance and regional perfusion. Complement depletion prior to endotoxemia resulted in preservation of normal systemic and hepatic perfusion. ATP and lipid peroxide levels were significantly abnormal in both groups of endotoxemic animals. Complement depletion alone did not significantly affect any of the variables studied. The maintenance of systemic and regional perfusion during endotoxemia was not cytoprotective implicating direct cellular injury independent of perfusion deficits in the pathogenesis of hepatic failure during endotoxemia. PMID- 8246497 TI - Methylprednisolone inhibits three classes of neutrophil receptors in human blood in vitro. AB - An analysis of receptor modulation on the neutrophil cell surface is essential for gaining insight into the activation and function of neutrophils in host defense. Moreover, agents that regulate the expression of surface receptors may have profound implications in the management of host response to infection. With this study, we extend previous observations in isolated cells of the putative ability of methylprednisolone sodium succinate (MPSS) to inhibit ligand binding to the formyl peptide receptor. Because neutrophils are exquisitely sensitive to isolation conditions, we have analyzed the regulation of receptor expression using flow cytometry in whole blood. This technique allows discrimination of neutrophils from other formed elements without isolation. Quiescent cells in blood exhibit low levels of formyl peptide receptor, CD11b/CD18, and CD14. We show that MPSS blocks upregulation of each of these receptors in response to three different stimuli (formyl peptide, lipopolysaccharide, and granulocyte macrophage colony-stimulating factor). The inhibition is reversible with an ED50 of approximately 0.4 mg/ml. From these observations, we conclude that the action of MPSS on neutrophils blocks a common response of receptors. Since these receptors probably function in part through independent signaling pathways, MPSS may function at a common site related to vesicular trafficking. Further investigation is needed to determine the specific means by which corticosteroids interfere with neutrophil upregulation mechanisms. PMID- 8246498 TI - Prevention of catheter-related infections by antiseptic bonding. AB - A novel catheter pretreated with the antiseptics chlorhexidine and silver sulfadiazine, designed to reduce catheter-related colonization and infection, was tested in both in vitro and in vivo studies. In vitro experiments demonstrated the long-lasting antibacterial properties of this catheter. For the in vivo study a total of 40 rats divided into different experimental groups were used. Colonization rates of both antiseptic bonded (AS) and control (C) catheters were assessed either three (-3) or seven (-7) days after implantation and local challenge using live Staphylococcus epidermidis ATCC 35984 with 10(7) colony forming units (cfu) per inoculum. At the time of removal, catheters, organ specimens, and blood samples were taken for cultivation. Significant reductions in the magnitude of colonization of the antiseptic catheters by the test organism were observed in all groups. The average number of cfu colonizing control segments exceeded those found on the treated catheter segments by log 3 (C3/AS3) and log 5 (C7/AS7), respectively (C3: 1.2 x 10(6) +/- 4.1 x 10(5) cfu/segment and AS3: 1.8 x 10(3) +/- 6.6 x 10(2); C7: 2.7 x 10(5) +/- 8.6 x 10(4) and AS7: 1.1 +/ 0.7; mean +/- standard error of the mean, all differences between matching groups statistically significant, Wilcoxon rank sum test, P < 0.0001). These data suggest that antiseptic catheters may substantially decrease the magnitude of catheter-related microbial colonization and subsequent catheter-related infections and may offer a more effective alternative to current methods. PMID- 8246499 TI - Free amino acids in biopsy specimens from the human colonic mucosa. AB - The content of free amino acids and total protein was determined in endoscopic biopsy specimens from the rectum, descending colon, transverse colon, and ascending colon in 10 patients. The amino acids were quantified by ion-exchange chromatography and were detected by fluorescence. The amino acid pattern and the rank order of the individual amino acids in the colon were different compared to those in plasma. Glutamate, taurine, aspartate, glutamine, glycine, alanine, serine, lysine, valine, and ornithine were the 10 most abundant amino acids in the colon. The rank order was the same in all of the different segments of the colon. The concentrations of the amino acids decreased the more aborally the biopsies were taken. The protein content in the rectum was significantly lower than that in the transverse colon, but there were no difference between the different segments otherwise. The study demonstrated the possibility of determining free amino acids from endoscopic biopsies of human colonic mucosa. Biopsy specimens from the descending colon and/or rectum/sigmoid colon may be considered representative of the entire large intestine. The technique may be used for repeated sampling in studies of the amino acid metabolism of the intestinal mucosa. PMID- 8246500 TI - Protection of cardiac myocytes from hypothermic injury by cardiac fibroblasts isolated from neonatal rat ventricle. AB - In this study, we evaluated the functional and biochemical effects of cardiac fibroblasts (CFs) on cardiac myocytes (CMs) incubated under hypothermic conditions. CMs and CFs were isolated from neonatal rat ventricles and cultures of myocytes only or myocytes in coculture with CFs were established. A CM and CF concentration of 2.5 x 10(5) cells/ml was chosen and the optimum proportion of CF was determined to be 33% (ml CM:ml CF) as follows: 6:0 (group C-a), 4:0 (group C b), 4:2 (group M-F). On the fourth day of culture, cells were incubated at 4 degrees C for 6, 12, 18, 24, 36 and 48 hr. After each incubation, creatine phosphokinase ((PK) and lactate dehydrogenase (LDH) were measured in the medium, and then the cells were cultured for an additional 24 hr at 37 degrees C to evaluate the recovery of the CM contracting rate. In groups C-a and C-b (n = 6 each), the recovery ratio of CM contracting rate decreased at 18 hr (C-a, 59.0; C b, 55.6% of control; i.e., contracting rate prior to hypothermic incubation), reaching null levels at 48 hr. However, group M-F (n = 6) showed a significantly increased recovery at 18 hr (95.7%, P < 0.05; at 48 hrs, 27.6%). Release of CPK and LDH in groups C-a and C-b increased gradually by 24 hr and showed a marked increase at 48 hr (C-a, 114.3 x 10(-2), 194.2 x 10(-2); C-b, 81.6 x 10(-2), 118.6 x 10(-2) IU/flask, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246501 TI - The fractal geometry of evolution. AB - This study is aimed at showing that the fractal geometry of taxonomic systems (Burlando, 1990) reflects self-similar evolutionary pattern. Evidence is achieved by three steps: (i) examination of taxonomic data from the fossil record; (ii) examination of taxonomic data from phylogenetic systematics; (iii) comparisons among different levels of the taxonomic hierarchy. In each step, all or nearly all the examined assemblages yield frequency distributions of numbers of subtaxa within taxa which fit a hyperbolic model function, confirming the fractal pattern. The first two steps show that the pattern is not deriving from classification bias, while the third one verifies the self-similarity of evolutionary radiations. According to the first and third step, self-similar cladogenesis consists in the arising of many isolated lineages and clumps of lines, the latter consisting of isolated lines and clumps, and so on. The properties of fractals led to the hypothesis that scaling diversity emerging from taxonomy could actually encompass the species level, thus limiting the importance of species within the evolutionary context in favour of a more comprehensive view of life diversification. PMID- 8246502 TI - The incorporation of gel pressure into the irreversible thermodynamic equation of fluid flow in order to explain biological tissue swelling. AB - After the Donnan osmotic potential derived theory of corneal stromal swelling was suggested (Hodson, 1971) it later was proposed, and verified experimentally, (Elliott et al., 1980) that the theory could be applied even in situations where there were no bounding cellular membranes attached to the corneal stroma. This was a paradox. The central problem with swelling in the absence of membranes (i.e. with a gel boundary whose reflexion coefficient is essentially zero) is that the driving force is not readily understandable as osmotic (but see Oster & Peskin, 1992) and yet the magnitude of the swelling fitted quantitatively with Donnan theory. We believe that we have resolved this paradox with a new and expanded irreversible thermodynamic relationship for fluid flows into and across biological tissues. In the course of the derivation, fresh concepts arise: for example the relationship of salt disparity is described which forbids diffusible salt-generated chemical and osmotic potentials to be simultaneously at equilibrium in the presence of ionised macromolecules and this relationship is developed to generate a new intrinsic thermodynamic property which is termed gel pressure and which drives fluid flow. Gel pressure provides a theoretical basis for biological tissue swelling. Microscopically, gel pressure is identified as the electrostatic potential developed by the mutual repulsion of the fixed matrix charges. PMID- 8246503 TI - Diffusion patterns on domains representing developing Xenopus retina. AB - The retinotectal map of mature Xenopus frogs has linear metrics. The regions of the Xenopus retina were modeled during embryonic and early larval development with a set of standard geometrical shapes. A disk was used to model the entire early retina, rings of various thickness were used to model the ciliary margin (the growth zone of the larval retina), and portions of a sphere were used to model the entire late embryonic and larval retina. Gradients produced by several reaction-diffusion pattern-generating configurations were considered: Gierer Meinhardt and Kauffman-type activator-inhibitor systems; an ideal Turing-like two morphogen system; a point source with diffuse sinks; and variously placed point source-point sink combinations. A few special arrangements produced linear metrics: a point source-point sink system produced linear angular metrics on a thin ring; and ideal Turing system produced linear cartesian metrics on a thin ring; and carefully configured multiple sources and sinks produced linear metrics on a disk. These cases could lead to linear map metrics, but are biologically implausible. Thus linear map metrics are probably not a direct consequence of linear positional values, and more generally, most embryonic tissues probably use non-linear gradients of positional information. Formation of linear map metrics may be functionally important for Xenopus frogs, accounting in part for the evolutionary preservation of mechanisms for interactions among neighboring retinal fibers. PMID- 8246504 TI - Quantitative study of the control of HIV-1 gene expression. AB - The biochemical processes that together determine the expression of a human immunodeficiency virus (HIV) provirus integrated into the genome of a host cell are translated into mathematical form to give a dynamic model that can be examined quantitatively. The model includes the contribution of cellular enhancers to transcription of the provirus, the subsequent splicing of the full length transcript, the nuclear export of transcripts and the translation of tat and rev mRNA to produce viral control proteins. The model is completed by the formulation of the feedback effects of Tat protein on transcription and translation and the feedback effect of Rev protein on nuclear export. Initial parameter values are estimated from available experimental data. The model is a formalization of a commonly accepted scheme, but one that has only previously been considered qualitatively. Quantification requires more explicit assumptions, but then allows the precise determination of resultant model behaviour and the stringent comparison of behaviour with experimental observation. Least squares matching of model behaviour to the observed appearance of mRNA in infected H9 cells gives very close agreement and, in particular, supports recent proposals concerning multimerization of the Rev protein. General quantitative characteristics of dynamic and steady-state behaviour of the model are determined and discussed; their possible contribution to the latency of HIV infection is considered. PMID- 8246505 TI - Tolerance to hormones and receptors in an idiotypic network model. AB - Using a simple mathematical model of the Jerne network, we investigate the conditions of antigen presentation that result in tolerance to pairs of antigens with complementary shapes, such as hormones and receptors. This study is motivated by the prevalence of auto-immune diseases involving hormones or neurotransmitters and their respective receptors as self-antigens. The model shows that, in order to ensure full tolerance to coupled antigens in those conditions that ensure tolerance to isolated antigens, both antigens have to be presented simultaneously in similar conditions. This result suggests a possibility for preventing some auto-immune diseases. PMID- 8246506 TI - Dynamics and control of bipedal locomotion. AB - The human frame is built for walking. It has both the right kinematics and the right dynamics--so much so, in fact, that our legs are capable of walking without any motor control. Their gait can be sustained simply by interaction of gravity and inertia, in a natural limit cycle which we call passive dynamic walking. This cycle needs motor input only for starting and stopping, for modulation when terrain calls for irregular strides, and for energy supply when the need arises. Analytical study reveals that any of several simple control strategies are effective for these purposes. This helps to explain why dextrous and efficient walking is so easy to a child to master. Moreover, it suggests that rehabilitation and robot design may be less difficult than one might at first imagine. PMID- 8246507 TI - An exact algebraic theory of genetic drift in finite diploid populations with random mating. AB - We study random mating in a finite diploid monoecious population with two alleles at one locus. We take into account selection and a constant probability of self mating. The generations are assumed discrete (non-overlapping). An exact algebraic theory of genetic drift is developed by the technique of transition probability matrices. The theory works on the genotype level (Hedrick, 1970), but also allows the population size to vary from generation to generation. Thus, it gives an exact statistical description of bottleneck processes, as an inhomogeneous Markov chain. Numerical Monte Carlo simulations are performed and give full agreement with the present algebraic theory. The decay rates for heterozygosity agree with the asymptotic theory by Wright (1931). PMID- 8246508 TI - Mechanisms of specificity in mRNA degradation: autoregulation and cognate interactions. AB - Autoregulation of gene expression is a common control mechanism for a large number of transcriptional units. Cases of self-regulation of the stability of various messenger RNAs (mRNAs) are re-evaluated here, and a general hypothesis for the origins and the mechanism of this process is presented. It is proposed that post-transcriptional autoregulation and mRNA stability are closely associated processes that might represent a general class of gene regulation mechanisms, with special regard to mRNA-protein cognate interactions. Generalizing from known examples, autoregulation is here considered to induce the decay of certain messenger RNAs through a yet undiscovered mechanism. Autoregulation via cognate interactions might be the vestigial process of a primitive world, where protein-nucleic acid interactions originated. The model can therefore serve as a framework to study the origins of the genetic code in particular, and gene expression in general. PMID- 8246509 TI - Analysis of striatal dynamics: the existence of two modes of behaviour. AB - The qualitative dynamical behaviour of a neural model based on the mammalian neostriatum was analyzed. The neostriatum was modelled as a mutually inhibitory network of physiological neurones, which was driven by excitatory afferents from the cerebral cortex. The analysis defined the conditions under which the system would enter into one of two dynamic modes, competition or co-activation, in terms of the parameters defining receptor-operated and voltage-sensitive channels in the neuronal membrane. We have previously argued that the mode of co-activation in the neostriatum may correspond to the state of muscular rigidity which occurs as a symptom of Parkinson's disease. The present work extends a preliminary analysis of a two-neurone system to a system of arbitrary size. An explicit prediction is made of the conditions under which a transition from co-activation to competition will occur, which is testable experimentally. The wavelength of a non-uniform activity pattern produced by small departures from uniform afferent drive is determined for one- and two-dimensional arrays of neurones. Two mild assumptions about the connectivity of the network were used to simplify the analysis, namely that the network was symmetric and homogeneous. The implications of departures from these assumptions for understanding the disordered movement seen in Huntington's disease are also considered. PMID- 8246510 TI - Effects of high frequency stimulation on cardiac tissue with an inexcitable obstacle. AB - We study numerically the effects of high-frequency stimulation in excitable cardiac tissue with an inexcitable obstacle using a Fitz-Hugh Nagumo type model. We show that if the frequency of stimulation is sufficiently high and the size of the inexcitable obstacle is sufficiently large, then a re-entrant pattern can be initiated. We also show that with overdrive stimulation a re-entrant pattern can be removed provided there are no obstacles at which new re-entrant patterns are created. PMID- 8246511 TI - Influence of diffusion and gravity on the adhesion of a two-component mixture of hard spheres on a flat surface. AB - The adhesion of hard spheres, modeling particles of biological interest (proteins, bacteria, cells), on flat surfaces is investigated by means of Monte Carlo simulations. These computations include the Brownian diffusion of the particles in the bulk fluid, as well as the systematic displacement due to the gravitational field. The size of the particles influences directly both diffusion coefficient and net weight, with the consequence that the coverage at the jamming limit depends on this parameter. Results obtained in a former paper based on a lattice model are confirmed by the present continuous space model. In order to gain a better understanding of the adsorption competition of two types of particles, the proposed model is applied to the case of binary mixtures of spheres. For polydispersed suspensions, various parameters determine the final coverage, as well as the distribution of the small and large particles on the surface: the radii of the particles and the respective proportions of them in the infinitely large reservoir from which they are randomly selected. In this way, it is shown that the chronology of the adhesion of the small and large particles strongly influences the final number of each type of spheres fixed on the surface. Qualitatively, the present results resemble those obtained with disks placed by means of a classical random sequential adsorption mechanisms. Quantitatively, however, the number densities and coverage values determined in this way are significantly different due to the inclusion of the gravity and of the diffusion in the model. PMID- 8246512 TI - The elements for a classification of units of genetic information with a combinatorial component. AB - An integrative approach to the study of the regulation of gene expression has been undertaken here. The main goal of this approach is to make explicit the common rules that govern the relative location of regulatory sites within operons and other units of genetic information (UGIs). A classification that emphasizes the regulatory properties of UGIs can be achieved by partitioning UGIs into short sequences with defined properties. Such a classification scheme can be precisely defined as a Grammar with a component of combinatorial (rewriting) rules, and a dictionary component. Sequences have then to be grouped into classes such that any sequence of the same class can mutually substitute and produce novel regulatable UGIs. It is shown here that individual nucleotides cannot define such classes--they are far from equivalent to phonemes. Neither pairs, triplets or any short sequence with a defined number of nucleotides can define productive substitutions. Defined sequences like promoter, operator and activator binding sites are the smallest elements of combinatorial rules within the defined range of transcription initiation of sigma 70 Escherichia coli promoters. PMID- 8246513 TI - Local and non-local receptor signalling. AB - It is suggested that the physiological response caused by an excited membrane bound receptor can be either local, i.e. localized close to the receptor site, or non-local, spreading over a large volume in the cell. A simple model is presented which shows how locality and non-locality are related to the intrinsic "efficacy" of the drug used in classical receptor theory. It is further demonstrated how the shape of the physiological response versus receptor occupancy is determined by the number of receptor sites which contribute to the physiological response at a given location in the cell. PMID- 8246514 TI - Entrainment of the respiratory rhythm: a new approach. AB - When we periodically perturbed the central respiratory oscillator with a controlled periodic stimulation provided by a mechanical ventilator, an entrainment phenomenon occurred: the actual rhythm of the respiratory centres was phase locked to the periodic stimulation. In some experiments, we observed an intermittence phenomenon: the respiratory rhythm successively seemed entrained, departed from this mode for few cycles, returned to the previous pseudo entrainment and so on. From intermittence data, we were able to plot a phase response curve. From literature data on the effects of a single stimulation applied at various moments in the respiratory cycle, we built up a mathematical model designed to simulate entrainment experiments. This model simulated entrainment phenomenon, but did not satisfactorily accommodate the experimental phase response curve. The section of the simulated phase response curve responsible for discrepancies was the result of the combination of an inspiratory shortening owing to a stimulation occurring during inspiration, and a positive relationship between inspiratory duration TI and expiratory duration TE. We changed the TE-TI relationship, assuming that the expiratory duration is determined not by the inspiratory duration but rather by the lung volume at the end of inspiration. The revised model was then able to exhibit almost all the qualitative properties previously noted in experiments. The significance in biological terms of the new TE-TI relationship, which is apparently incompatible with almost all previous experimental data, is discussed. PMID- 8246515 TI - A shifting specificity model for enzyme catalysis. AB - A new model for general enzyme catalysis challenges the idea that transition state complementarity of enzyme active sites to the reactions they catalyze is the sole source of their catalytic efficacy. The "shifting specificity" model rejects the widely held view that strong interactions of the enzyme with substrate inhibit catalytic efficiency and previously published data are presented which show that a strong interaction of substrate with the enzyme active site actually facilitates its conversion to product. Furthermore, this new model defines a role for the entire enzyme molecule unlike most theories of enzyme catalysis which are concerned only with the interaction of the active site with substrate. This shifting specificity model for general enzyme catalysis may be expressed succinctly as: (i) enzymes have evolved to bind substrates; (ii) enzyme/substrate complexes have evolved to bind transition states; (iii) a stronger interaction of substrate with the enzyme facilitates a more rapid conversion to product. This last effect results from a more efficient modulation of the global enzyme conformation by tight-binding substrates. It is suggested that the addition of atomic mass to the enzyme, which interacts with the enzyme in the same manner in which the enzyme domains interact with one another, must alter the low-frequency, global vibrations of the enzyme to produce a different overall conformation. Nature has selected for that conformational change which shifts the active site complementarity from substrate-specific to transition state-specific. Thus, this model suggests a means for an efficient realization of the substrate-binding energy in the transition state of the reaction. General aspects of this model are discussed in light of the current view of enzyme catalysis. PMID- 8246516 TI - Seeking the evolutionary regression coefficient: an analysis of what comparative methods measure. AB - Two alternative classes of comparative statistical method differ in the way that the comparative data are used to test for an association between two quantitative traits. Directional comparative methods use reconstructions of the ancestral character states to calculate the changes between ancestral and descendant conditions along the branches of the phylogenetic tree. The set of changes in two or more traits is used to test for evidence of correlated evolution. Cross sectional techniques do not estimate changes along the branches of the tree, but rather make comparisons across the tips of a phylogeny, or between pairs of extant taxa (or between their higher nodes). These methods, then, study the association between pairs of traits representing the contemporary endpoints of evolution. The best known of the cross-sectional techniques, the species regression, simply regresses the species values of one variable onto those of another. However, it is shown here analytically that directional and cross sectional methods, despite making very different use of the data, estimate precisely the same evolutionary parameter: the association between the changes in two variables along the branches of the phylogenetic tree. Thus, comparative statistical techniques are able to recover the historical trends of evolution, that is, the ways in which evolution has proceeded along the branches of the phylogenetic tree, from analysis of the variation among the contemporary species of a phylogeny. This means that the choice between the two alternative traditions of comparative study cannot be based upon what the different methods purport to measure, but rather must be based upon the statistical properties of particular methods. In the light of this result, it is discussed here whether there are statistical reasons to prefer some methods over others. PMID- 8246517 TI - Sequestration of origins of chromosome replication in Escherichia coli by lipid compartments: the pocket hypothesis. AB - After initiation of chromosome replication in Escherichia coli, the newly replicated, hemi-methylated origins of replication are exempt from both re initiation and methylation for up to a third of the generation time. In the model proposed here, the exemption from methylation is attributed to the phospholipid preferences of proteins binding hemi-methylated origins to membrane; these proteins associate with unsaturated phosphatidylethanolamine which forms structures with small radii of curvature; hence, a lipid compartment or "pocket" forms around the origin making it inaccessible to cytoplasmic enzymes such as Dam methyltransferase. Since autolysins and peptidoglycan-synthesizing enzymes are sensitive to lipid composition, peptidoglycan synthesis is stimulated around this compartment and creates structures obtained in membrane fractionation that specifically bind hemi-methylated origins. PMID- 8246518 TI - Characterization of myocardial stress-length relations. AB - For several decades, some muscle physiologists, cardiac physiologists and cardiologists have normalized systolic load-dimension relations to a positive strain reference dimension such as the muscle length which is optimal for stress development or the chamber volume resulting in normal end-diastolic stress. Some have viewed isometric stress development at reference length as a contractility index, and some have viewed shortening from reference length in the absence of afterload as an important property (shortening ability or mobility). These intuitive choices together constitute a coherent elastic theory which is more appropriate for activated muscle than is the classical theory, wherein slack length is the reference and slope of the stress-strain relation is the main parameter. The stress-length relation is readily modeled by functions in which contractility and shortening ability are parameters, entirely separate from each other and from those representing shape of the load-dimension relation. PMID- 8246519 TI - A fast algorithm for the optimal alignment of three strings. AB - Ukkonen's (pair-wise) string alignment technique is extended to the problem of finding an optimal alignment for three strings. The resulting algorithm has worst case time-complexity O(nd2) and space-complexity O(d3), where the string lengths are n and d is the three-way edit-distance based on tree-costs. In practice, the algorithm usually runs in O(n + d3) time. The algorithm is particularly fast when the strings are similar, in which case, d << n. Three-way alignment is an important special case in string alignment. Each internal node in an unrooted, binary evolutionary-tree has three neighbours. The algorithm presented can be used as an iterative step in a heuristic multiple-alignment program for more than three strings. PMID- 8246520 TI - Studies on a recent class of network models of the immune system. AB - It is argued that the realism of computer simulations of network models of the immune system depends basically on the coherence of these models with the essentials of the known physiology of the cells and molecules selected to be modelled and on the incorporation in them of the different compartments of activated B cells. Focusing on these two aspects, here we analyse the simplifications and assumptions that go implicit in the formulation of a recently developed new class of network models that distinguish between immunoglobulins and B cells. This is approached by first building a general model which incorporates explicitly the kinetics of different B-cell compartments as well as a splenic compartment and a peripheric one for immunoglobulins, and then formally studying the simplifications on this model that are necessary to recover the initial simpler models. Following this procedure, it is shown that the effective coefficients of the different rate terms in the simpler models are particular combinations of the elementary rates obtained empirically. These relations reflect the particular assumptions associated with each simplification step. Also, it is shown that the usual biological interpretation of some of the coefficients in the ordinary differential equations of the simpler models is inconsistent with the more exact general model, unless one makes certain unreasonable assumptions about B-cell physiology. The relevance of this approach in providing variables with a biologically identifiable reality and for realistic, testable, computer simulations is discussed. PMID- 8246521 TI - Simulation studies of self-replicating oligoribotides, with a proposal for the transition to a peptide-assisted stage. AB - A two-substrate Michaelis-Menten mechanism previously proposed for the self replication of RNA-like oligomers is developed. Differential growth depends on the existence of two pairs of complementary monomers and leads to 2n groups of 2n components each (n is the oligomer size). As n increases the 2n groups tend to overlap with one another, and the efficiency of the process to increase the information content of the strands decreases. In a second stage we suppose that randomly synthesized peptides with one predominant amino acid interacted with the ribotides, increasing the growth rate of some of them, and at the same time had their mean life increased by interactions with other ribotides of the same kinetic group. Natural selection could have preserved a favourable codon anticodon-amino acid correlation, the precursor of the modern genetic code. PMID- 8246522 TI - Speculations on the roles of ADP-ribosyl transferase based on analogies between RecA and poly(ADP-ribose) polymerase. AB - A model for eukaryotic DNA damage repair is proposed in which poly(ADP-ribose) polymerase(NAD+ ADP-ribosyl transferase, EC 2,4,2,30) plays an important role. In this model, poly(ADP-ribose) polymerase regulates transcription of genes that are induced by DNA-damaging agents. This transcriptional regulation results from poly(ADP-ribosyl)ation and inactivation of DNA sequence-specific regulatory proteins such as silencer element-binding proteins, thereby inducing transcription of DNA polymerase beta, which is a DNA repair enzyme in higher eukaryotes. Poly(ADP-ribose) polymerase has a number of similarities to RecA in Escherichia coli. Therefore, the genes related to DNA damage repair in higher eukaryotes are proposed to form a "poly(ADP-ribose) polymerase regulatory network" similar to the "SOS regulatory network" in E. coli. PMID- 8246523 TI - Spread of phenotypic plasticity or genetic assimilation: the possible role of genetic constraints. AB - This paper considers possible outcomes of evolution in sexual diploid populations subjected to frequency-independent selection in favour of a phenotype which has appeared as a consequence of a change in environmental conditions. Depending on the details of genetic determination for the different development strategies ("plastic" or "canalized"), this situation results at least in either (i) the invasion of the population by "plastic" strategists, whose developmental plasticity was revealed by the environmental change, or (ii) the genetic assimilation of the new phenotype, or (iii) intermediate states in which only a certain proportion of individuals is genetically assimilated at equilibrium. Spread of plasticity during evolution in a series of distinct environments might, therefore, be hindered by genetic constraints. PMID- 8246524 TI - A note on the reduction of the dynamics of multilocus diploid genetic systems with multiplicative fitness. AB - The dynamics of any diploid multilocus genetic system with two alleles and uniform dynamics per locus can be deduced from that of the corresponding single locus system by trinomial sampling to produce the frequencies of the different mutant classes, provided that fitness is multiplicative, and there is linkage equilibrium (allowing for physical linkage, however). The component processes that can be considered are mutation, gene conversion, gamete production (with recombination) and random mating, and selection. If mutations occur according to Bernoulli trials, outcrossing sexual systems have a genetic load exactly deducible from that of the single-locus system. PMID- 8246525 TI - Overdispersed molecular evolution in constant environments. AB - According to recent data analysis of DNA sequences, the dispersion index, defined as the variance-to-mean ratio of the number of base substitutions in a lineage, is often much larger than unity, which is in conflict with simple Poisson processes assumed in the molecular clock hypothesis. In this paper, it will be shown that the dispersion index can be much larger than unity in a model in which the fitness of DNA sequences, mutation rate, and population size are all constant with time, and that moderately deleterious sequences are more abundant than the best fit sequences. Since the fixation probability of novel mutations depends on their fitness relative to the current sequence, the neutral mutation rate is enhanced once a deleterious mutation is fixed. In a simple case with two fitness classes, a large dispersion index can be produced by moderately deleterious mutations (3 < 4Ns < 6), but neither by nearly neutral (4Ns < 2) nor by strongly deleterious (4Ns > 7) mutations. Analysis of the case with 100 fitness classes shows that the dispersion index is insensitive to the population size, but greatly changes with the fitness distribution of DNA sequences. The model can explain why non-synonymous substitutions often have a larger dispersion index than synonymous substitutions. PMID- 8246526 TI - Dimensionality, time scales and membrane thermodynamics. PMID- 8246527 TI - Angolan medicinal plants used also as piscicides and/or soaps. AB - A short description of some medicinal plants found during a 10-year ethnological study of traditional medicine in Angola. The point of interest is the double use of these plants: they are employed as drugs and as ichthyotoxics or soaps. PMID- 8246528 TI - Species identification and chemical analysis of psychoactive fungi in the Hawaiian islands. AB - Several fungi species collected in the Hawaiian Islands have been reported to be psychoactive. Previous chemical analyses together with the present study indicate that 5 coprophilous and one non-coprophilous species occurring in the islands are now known to contain psychoactive alkaloids. At least some of these species are consumed in the Hawaiian Islands, as well as elsewhere, for non-traditional, recreational purposes. These include Copelandia cyanescens (Berk. et Br.) Singer, Copelandia tropicalis (Ola'h) Singer and Weeks (syn. Panaeolus tropicalis Ola'h), Copelandia anomala Murrill, and Panaeolus subbalteatus (Berk. and Br.) Sacc., which have already been described from the Hawaiian Islands. Three more mind altering fungi and one non-psychoactive species are reported from this archipelago for the first time. These psychoactive fungi include Copelandia bispora (Malencon et Bertault) Singer and Weeks from O'ahu, Copelandia cambodginiensis (Ola'h et Heim) Singer and Weeks from O'ahu, and Amanita muscaria (L.) Hooker from Kaua'i. Panaeolus goossensiae Beeli identified from O'ahu contains tryptamine compounds; however, the psychoactive alkaloids psilocybin and psilocin were not found in this dung species. PMID- 8246529 TI - Alstonia species: are they effective in malaria treatment? AB - A review of the literature on Alstonia species indicates that evidence in support of their effectiveness in the treatment of malaria is controversial. The antiprotozoal activity of the major alkaloid present in Alstonia species, echitamine, was assessed in vitro against Plasmodium falciparum and Giardia intestinalis. Echitamine displayed little antiplasmodial activity, but two quinoline alkaloids from A. coriacea (corialstonine and corialstonidine) were found to have some activity against P. falciparum although this was approximately 10 times less than that of quinine. None of the three Alstonia alkaloids was active against G. intestinalis. These results are discussed in the context of previously published data. PMID- 8246530 TI - Antimalarial activity from 'Mhekara' (Uapaca nitida Mull-Arg.), a Tanzanian tree. AB - An aqueous decoction of the root bark of Uapaca nitida Mull-Arg. is currently used locally at the Benedictine Mission at Peramiho in Tanzania to treat malaria. We have now demonstrated that extracts of root bark and leaves of this tree are active against the multidrug-resistant K1 strain of Plasmodium falciparum in vitro. An ethanolic extract of root bark showed activity against P. berghei in mice but at a dose which also showed toxic effects. The use of this plant in treating malaria appears to be novel and further studies would be of value. PMID- 8246531 TI - Tapping an Amazonian plethora: four medicinal plants of Marajo Island, Para (Brazil). AB - This study focused its attention on four medicinal plants (Carapa guianensis, Elephantopus scaber, Piper umbellatum, Stachytarpheta cayenensis) used by Caboclo communities on Marajo, the main island of the Amazon delta. In the field, interviews were conducted with Caboclos and the medicinal usages and preparation procedures of the four plants were recorded. In the laboratory, the plant extracts were subjected to bioassays and their crude chemical composition was established. All four plants showed significant bioactivity and the chemical tests confirmed the presence of bioactive compounds. In addition, the results of both the field and laboratory studies corresponded well with those of a literature search. The ethnopharmacological significance of the four plants is discussed. PMID- 8246532 TI - Cytokine and complement levels in patients undergoing cardiopulmonary bypass. AB - Patients undergoing cardiopulmonary bypass are known to develop whole body inflammation that often results in a characteristic syndrome early postoperatively. This phenomenon has been attributed to complement activation caused by exposure of blood to the foreign surfaces of the cardiopulmonary bypass circuit. It has been unknown if cytokines are involved. Plasma levels of complement activation products (C3a, C4a, C5a, and C5b-9), interleukins (IL-1 beta, IL-2, IL-4, and IL-6), and tumor necrosis factor-alpha were measured at multiple time points before, during, and after cardiopulmonary bypass in 29 patients. No significant increase over preinduction levels was seen in the cytokines except for IL-6, which was significantly increased during cardiopulmonary bypass (p < 0.001), reaching a maximum 3 hours after cardiopulmonary bypass. C3a, C4a, and C5b-9 levels were significantly elevated during cardiopulmonary bypass (p < 0.001), with maximum C5b-9 levels preceding the IL-6 elevation. Heparin coating of the cardiopulmonary bypass circuit was not demonstrated to have an effect on activation of complement or cytokine production. There was no statistically significant correlation among hemodynamic variables or pulmonary function and complement, interleukin, or tumor necrosis factor-alpha levels. These results confirm the presence of complement activation and demonstrate the production of IL-6 after the generation of C5b-9 in patients undergoing cardiopulmonary bypass. IL-6 may contribute to adverse systemic reactions associated with cardiopulmonary bypass. PMID- 8246533 TI - Tissue-type plasminogen activator and fibrin monomers synergistically cause platelet dysfunction during retransfusion of shed blood after cardiopulmonary bypass. AB - Reduced hemostasis and bleeding tendency after cardiopulmonary bypass results from platelet dysfunction induced by the bypass procedure. The causes of this acquired platelet dysfunction are still subject to discussion, although, recently, greater emphasis has been placed on an overstimulated fibrinolytic system as a probable cause. In the first part of this study we assessed the effects of postoperative retransfusion of shed blood on blood loss to patients undergoing cardiopulmonary bypass. We observed that increasing concentrations of fibrinogen degradation products and tissue-type plasminogen activator stimulating activity in shed blood correlated significantly with a higher postoperative bleeding tendency (p < 0.05 for both). We further noted that retransfusion of shed blood increased the total postoperative blood loss by 43% (925 versus 1320 ml, p < 0.05). On the basis of these clinical observations, we hypothesized that the increased bleeding tendency was caused by fibrinolysis. In the second part of this study we collected evidence in support of this hypothesis by an in vitro study, in which we introduced similar (pro)fibrinolytic activity to platelet-rich plasma and measured the influence of this treatment on platelet function indicated by ristocetin agglutination. Tissue-type plasminogen activator and fibrin monomers (tissue-type plasminogen activator stimulator) together induced severe platelet damage, resulting in a decreased ristocetin agglutination response. Therefore, we propose a fibrinolysis-related mechanism for platelet dysfunction during cardiopulmonary bypass, dependent on fibrinolytic factors such as fibrin monomers, D-dimers, and tissue-type plasminogen activator. PMID- 8246534 TI - Profound systemic hypothermia protects the spinal cord in a primate model of spinal cord ischemia. AB - Spinal cord ischemia with resultant paraplegia or paraparesis remains an important clinical problem after operations on the thoracoabdominal aorta. Because hypothermia has a protective effect on ischemic neural tissue, we developed a baboon model of spinal cord ischemia to simulate the situation encountered clinically for resection of aneurysms of the thoracoabdominal aorta and to determine whether profound hypothermia produced by hypothermic cardiopulmonary bypass has a protective effect on spinal cord function. After cardiopulmonary bypass was established, the aorta was clamped distal to the left subclavian artery and proximal to the renal arteries for 60 minutes. Group I animals (n = 9) underwent aortic clamping at normothermia (37 degrees C), and group II animals (n = 9) were cooled to a rectal temperature of 15 degrees C before aortic clamping and underwent cardiopulmonary bypass at this temperature until the aorta was unclamped. Of the eight operative survivors in group I, six animals were paraplegic and two were paraparetic, whereas all six group II animals that survived the procedure were neurologically intact (p = 0.0002). The protective effect of hypothermia was associated with blunting of the hyperemic response of spinal cord blood flow (determined by the radioactive microsphere technique) in the lower thoracic and the lumbar segments of the spinal cord after unclamping of the aorta. Profound hypothermia produced by hypothermic cardiopulmonary bypass may be an effective method of protection of the spinal cord in patients undergoing repair of aneurysms of the thoracoabdominal aorta and may reduce the prevalence of ischemic injury to the spinal cord. PMID- 8246535 TI - Right ventricular rupture. A complication of postoperative mediastinitis. AB - Mediastinitis-related right ventricular rupture is an unusual but potentially life-threatening complication of cardiac operations. Between January 1981 and December 1990, a total of 10,182 patients underwent heart operations for ischemic, valvular, and congenital heart disease at the Montreal Heart Institute. Forty-eight patients (0.5%) had postoperative mediastinitis necessitating surgical exploration and sternal debridement. The mediastinum was left open for daily irrigation with povidone-iodine and chest reconstruction was postponed. During treatment, seven patients (0.07%) had right ventricular rupture necessitating immediate surgical repair. All had ischemic heart disease before the operation. There were five women and two men, ages ranging from 52 to 65 years (mean 58 +/- 5 years). Surgical repair consisted of autologous patch covered with omentoplasty assisted with cardiopulmonary bypass. Two patients died, one during the operation of massive hemorrhage and the other 10 days after the operation of uncontrolled sepsis. Five patients survived 2 to 29 months (mean 23 +/- 10 months) after right ventricular rupture, with an overall survival of 71%. Obesity was more frequent in the patients with right ventricular rupture and was found to be a significant risk factor (multivariate analysis, p < 0.05, relative risk 3.22). Histologic examination of the right ventricle in the patient who died after a successful repair revealed fatty infiltration of the right ventricular wall. This may have predisposed the patient toward ventricular rupture. In conclusion, right ventricular rupture, an unusual event in heart surgery, is related to open sternal debridement. Favorable outcome of this complication depends on immediate surgical management, autologous repair, and the use of omentoplasty. PMID- 8246536 TI - Nonthoracotomy lead system for implantable defibrillator. AB - Over a 2-year period, 110 patients underwent attempted implantation of an automatic cardioverter-defibrillator using the nonthoracotomy lead system. Indications included sustained monomorphic ventricular (n = 62), nonsustained with poor ventricular function (n = 7), ventricular fibrillation (n = 21), ventricular tachycardia/fibrillation (n = 18), and familial long QT syndrome (n = 2). There were 90 male and 20 female patients. Mean age was 57 +/- 15 years. Sixty percent had previous coronary bypass or valve operations, or both. Mean left ventricular ejection fraction was 30% +/- 14%, cardiac index was 2.4 +/- 0.9 L/m2, and systolic pulmonary artery pressure was 41 +/- 14 mm Hg. Under general anesthesia, the nonthoracotomy lead was introduced through the left subclavian vein. The subcutaneous patch and generator were placed posteriorly on the serratus muscle and left upper quadrant, respectively. The length of the procedure was 116 +/- 44 minutes and the mean number of defibrillation shocks for a successful implant was 8 +/- 4. Eighty-five patients (77%) had successful implantations. Failures were due to high defibrillation threshold (n = 23) and inability to place a right ventricular lead (n = 2). Predictors of failure included preoperative antiarrhythmic drugs and cardiac index of 1.8 +/- 4 L/m2 or less (p = 0.004). Three patients (2.7%) died after the operation of heart failure (n = 2) and chronic heart transplant rejection (n = 1). Complications included lead migration or dislodgment (n = 8), infection (n = 1), and hematoma (n = 3). In summary, the nonthoracotomy lead system may provide an alternative in patients undergoing cardioverter-defibrillator implantation. PMID- 8246537 TI - Video-assisted thoracoscopic stapled wedge excision for indeterminate pulmonary nodules. AB - Between June 1991 and July 1992, 118 patients (57 men and 61 women) underwent video-assisted thoracoscopy for indeterminate pulmonary nodules. Median age was 64 years (range 30 to 85 years). Thoracotomy was performed in 33 patients (28.0%) after thoracoscopy only because the nodule could not be located in 17 patients, was too large to safely resect in 5, appeared malignant in 4, and for technical reasons in 7. Eighty-five patients underwent thoracoscopic wedge excision. Twenty one (24.7%) of these 85 patients also had thoracotomy--15 to perform formal lung resection for bronchogenic carcinoma, 3 for nondiagnostic abnormalities, 2 to locate a second nodule, and 1 for stapler malfunction. The remaining 64 patients (54.2%) had only video-assisted thoracoscopic wedge excision. A single wedge excision was performed in 56 patients, two in 6, and three in 2. Pathologic examination of these 74 nodules revealed a granuloma in 30, metastatic cancer in 25, hamartoma in 7, lymphoma in 1, and other benign lesions in 11. There were no deaths and only 4 (6.3%) complications in these 64 patients. The 64 patients treated by thoracoscopy only were compared with a similar group of 64 patients who had wedge excision via thoracotomy without prior thoracoscopy. Postoperative analgesic requirements were less in the patients treated by thoracoscopy. Median hospitalization in the thoracoscopy group was 3 days compared with 6 days in the thoracotomy group (p < 0.05). Median total charge for the thoracoscopy-only group was $12,898 as compared with $12,502 for patients undergoing wedge excision via thoracotomy. We conclude that thoracoscopic wedge excision is a safe and effective procedure in selected patients with an indeterminate pulmonary nodule. A significant number of patients (45.8%), however, required a thoracotomy to accomplish a safe operation or to ensure adequate staging and resection for malignancy. Although thoracoscopy reduces postoperative analgesia requirements and shortens hospital stay, total hospital charges were similar to charges for a wedge excision via thoracotomy. PMID- 8246538 TI - Malignant disease appearing late after operation for T1 N0 non-small-cell lung cancer. The Lung Cancer Study Group. AB - Occurrences of malignancy in 308 patients who were clinically free of cancer 60 months or more after surgical treatment of T1 N0 non-small-cell lung cancer are summarized. At last report, 210 patients remained alive with no evidence of malignant disease, 43 patients died of nonmalignant causes, and 55 patients had 59 occurrences of malignant disease. Late lung cancer recurrence was observed in 22 patients (concurrent with nasopharyngeal cancer in one patient and with laryngeal cancer in one patient). Metachronous second lung cancer was noted in 20 patients (including concurrent colon cancer in one patient and metastatic recurrence in one patient). Other nonpulmonary malignant tumors appeared in 13 patients. Including second lung cancer, 25 cancers of aerodigestive epithelium were observed in 23 patients. The malignancy-free survival advantage for patients with squamous cancer observed until 60 months after resection was not sustained at 60 months and beyond. At the time of last follow-up, 84 patients (27%) had died; 43 were free of malignancy, and 41 had malignancy (14 patients were alive with malignancy and 210 with no evidence of disease). These data reinforce two conclusions: (1) The probability of lung cancer recurrence or appearance of new lung cancer 5 years or more after successful operation in this select subpopulation of patients with lung cancer is of concern. (2) The malignancy-free survival advantage of patients with squamous cancer disappears after 5 years. PMID- 8246539 TI - Effect of preoperative irradiation on wound healing after bronchial anastomosis in mongrel dogs. AB - To determine the endurable radiation dose before bronchial anastomosis, the bronchial circulation of mongrel dogs was assessed after bronchoplastic surgery conducted after various single doses of radiation. Plastic surgery of the right main bronchus was carried out 1 week after cobalt 60 irradiation to the right hilar region. Bronchial blood flow was measured with laser Doppler velocimetry 7, 14, and 30 days after operation. Animals were killed 30 days after operation and the region of the bronchial anastomosis was examined histologically. The animals were divided into five groups: group A (control group without irradiation), group B (8 Gy irradiation), group C (16 Gy), group D (19 Gy), and group E (24 Gy). Wound healing of the anastomosed bronchus was excellent in groups A and B. In group A, bronchial blood flow did not drop below 80% of the preoperative level throughout the postoperative observation period. In group B, bronchial blood flow on day 7 after operation was only 65% of the preoperative level, but blood flow had returned to the preoperative level by day 30. In groups C, D, and E, bronchial blood flow was less than 60% of the preoperative level on day 30. There was a high prevalence of radiation-induced morbidity in the region of anastomosis in groups C, D, and E. Postoperative recovery of bronchial blood flow and wound healing of the anastomosed bronchus were thus delayed in dogs that had received high doses of radiation before operation. We conclude that the safe dose of preoperative radiation in clinically used fractionation is 36 Gy or less (corresponding to a single dose of 16 Gy or less in dogs). PMID- 8246540 TI - Immunoreactive thymosin alpha 1 in human thymus and thymoma. AB - Thymosin alpha 1-like immunoreactivity was assessed in human thymus and thymoma tissue extracts by means of a new radioimmunoassay that included an anti-thymosin alpha 1 mouse monoclonal antibody. Thymosin alpha 1-like immunoreactivity levels decreased with age in normal thymuses but not in thymomas. The average thymosin alpha 1-like immunoreactivity level was 45.0 +/- 52.1 ng/mg protein in normal thymuses and 273.9 +/- 205.0 ng/mg protein in thymomas. The average thymosin alpha 1-immunoreactivity level in thymomas was higher than that in normal thymuses. Thymosin alpha 1-like immunoreactivity levels in thymomas appeared to have no relationship to the clinical stage of the thymoma or associated diseases. When viewed according to histologic characteristics, the average thymosin alpha 1 like immunoreactivity level in polygonal cell thymomas (382.5 +/- 192.6 ng/mg protein) was significantly higher than that in the spindle cell thymoma (101.8 +/ 81.2 ng/mg protein). When viewed according to the degree of lymphocyte infiltration, thymomas could be classified according to four grades: absent, scant, moderate, and predominant. In predominant or moderate thymomas, the average thymosin alpha 1-like immunoreactivity level was higher than that in scant or absent thymomas. Also, thymosin alpha 1-like immunoreactivity levels in thymuses of patients with myasthenia gravis were relatively higher than those in patients with normal thymuses. PMID- 8246541 TI - Right-to-left interatrial shunt in rats with progressive pulmonary hypertension. AB - A right-to-left interatrial shunt may prolong survival in patients with pulmonary hypertension presumably because of decompression of the right side of the heart. To test this hypothesis, 74 rats with monocrotaline-induced pulmonary hypertension were followed up weekly with cardiopulmonary exercise testing with a metabolic treadmill system for exercise tolerance, heart rate, oxygen uptake, carbon dioxide production, and survival until subsequent or induced death 8 weeks after monocrotaline treatment. In rats with an interatrial shunt, oxygen uptake and carbon dioxide production were higher and survival was better (n = 22, 27%) than those in rats without a shunt (n = 52, 0%; p < 0.05). For the prospective assessment of the effects of a reversed shunt, 24 other rats underwent a left superior vena cava-to-left atrial appendage anastomosis as a functional interatrial shunt (atrial septal defect group) 4 weeks after monocrotaline treatment when severe pulmonary hypertension had developed and were compared with an additional 25 rats receiving a sham operation. Both groups had exercise capacity depressed to the resting levels by 2 weeks after operation. Although transcutaneous oxygen levels decreased in response to exercise in the atrial septal defect group, uptake and carbon dioxide production stayed higher than those in the sham operation group with significantly better survival 4 weeks after operation (atrial septal defect 30% versus sham operation, 0%; p < 0.05), at which time a reversed shunt was determined with systemic embolization of intravenously infused microspheres. A right-to-left interatrial shunt, anatomic or functional, preserved basal metabolism and prolonged survival in rats with progressive pulmonary hypertension. PMID- 8246542 TI - Maximum length of tracheal autografts in dogs. AB - When a primary anastomosis of the trachea is not feasible, grafting is required. We assessed the maximum length of tracheal autografts in 25 dogs. We used the autografts in this experimental study to avoid the immunologic complexity of allografts. Sections of cervical trachea ranging in length from 2.4 cm (6 rings) to 5.9 cm (14 rings) were excised and reimplanted as autografts. To increase the blood supply, we wrapped each graft in the greater omentum, which was brought up through the diaphragm. Grafts longer than 4.0 cm (8 to 10 rings) frequently showed stenosis, dissolution, or both in the middle part of the graft as a result of ischemia in that area, as demonstrated by laser blood flowmetry and injection study. We conclude that there is a limit to the length of a tracheal autograft, even if the blood supply has been increased by wrapping with omentum. PMID- 8246543 TI - Management of delayed esophageal perforation with mediastinal sepsis. Esophagectomy or primary repair? AB - Ninety patients with esophageal perforations were operated on at our institutions between 1970 and 1992. Thirty-four of them were seen after delayed diagnosis (> 24 hours) with mediastinal sepsis caused by perforation of the thoracic esophagus. There were 18 patients with spontaneous ruptures, 11 with instrumental perforations (including one caused during laparotomy), and 3 perforations caused by foreign bodies. One patient had perforation of an esophageal ulcer into the pericardium and another had perforation of an esophageal diverticulum into the mediastinum. Nineteen patients underwent primary repair of the perforation with cleansing and drainage of the mediastinum and the pleural cavity. The remaining 15 had primary extirpation of the thoracic esophagus, irrigation of the mediastinum with antibiotics, cervical esophagostomy, gastrostomy, and drainage of the mediastinum and pleural cavity. Nineteen of the 34 patients survived (hospital mortality 44%). Of patients with primary repair, only six survived (in hospital mortality 68%), whereas only two patients treated with esophagectomy died (in-hospital mortality 13%). The difference was highly significant (p = 0.001). The most common cause of death was multiorgan failure resulting from sepsis. Postoperative complications developed in four patients treated with primary repair (two sepsis, one empyema, and one anuria) and in seven patients treated with esophagectomy (two empyema, two sepsis, one pneumonia, one mediastinal abscess, and one brain abscess). After healing of the mediastinitis, the esophagogastric continuity was reconstructed with colon in 11 patients and stomach in two patients. In the management of delayed esophageal perforation with mediastinal sepsis, esophagectomy is superior to primary repair alone, which often leads to mediastinal leakage, continued sepsis, and death. PMID- 8246544 TI - Prognostic significance of pleural lavage cytology immediately after thoracotomy in patients with lung cancer. AB - Pleural lavage cytology immediately after thoracotomy was performed in 467 patients with lung cancer who had little or no pleural effusion. Forty-two patients (9.0%) had positive results. The positivity of pleural lavage cytology was significantly related to the degree of pleural extension of the tumor, microscopic pleural dissemination, cytologic results of minimal pleural effusion, pathologic stage, presence of lymphatic permeation or vascular invasion, and cell type (adenocarcinoma was predominant). The 3-year survival of the patients having negative and positive results of cytology were 68.7% and 22.9%, respectively. The prognosis of the group with positive results was as poor as that of patients with stage IIIB or IV disease. Pleural lavage cytology is an important prognostic factor that indicates microscopic exfoliation of cancer cells into the pleural cavity, that is, subclinical malignant pleural effusion. PMID- 8246545 TI - Relation between bronchoscopic findings and tumor size of roentgenographically occult bronchogenic squamous cell carcinoma. AB - A total of 105 lesions in 98 patients with roentgenographically occult bronchogenic squamous cell carcinoma were examined. The relationship of bronchoscopic findings to the depth of invasion into the bronchial wall and the length of longitudinal extension along the bronchus was documented. From viewpoints of the degree of difficulty of bronchoscopic detection and with reference to the height of the lesions, the bronchoscopic findings were classified into three categories: remarkable, minute, and hidden. Of the 105 lesions, 55 (52%) were remarkable, 27 (26%) were minute, and the remaining 23 (22%) were hidden. Of the 23 hidden lesions, 12 were within and 11 were beyond the range of endoscopic visibility. The maximal depth of bronchial invasion (mean +/- standard error) was 3.07 +/- 0.40 mm in the category designated remarkable and 1.62 +/- 0.47 mm in the category designated minute. The depth was 0.93 +/- 0.36 mm in the hidden lesions within the range of endoscopic visibility and 0.78 +/- 0.21 mm in the hidden lesions beyond the range of endoscopic visibility. The maximal length of longitudinal extension along the bronchus was 19.6 +/- 1.5 mm in the remarkable lesions, 9.9 +/- 1.4 mm in the minute lesions, 5.5 +/- 1.0 mm in the hidden lesions within the range of endoscopic visibility, and 8.6 +/- 2.1 mm in the hidden lesions beyond the range of endoscopic visibility. It is useful for predicting the depth of invasion to classify bronchoscopic findings into these three categories for the study of roentgenographically occult bronchogenic squamous cell carcinomas. PMID- 8246546 TI - What are the risk factors for arrhythmias after thoracic operations? A retrospective multivariate analysis of 267 consecutive thoracic operations. AB - Two hundred sixty-seven operations performed consecutively at the National Cancer Center Hospital, Tokyo, in 1990 were reviewed to define the prevalence, type, clinical course, and, especially, risk factors for arrhythmias after thoracic operations. Arrhythmias were identified in 63 operations (23.6%) and were more prevalent in several subgroups of patients than in others--those with lung cancer, pneumonectomy, mediastinal lymph node dissection, and those older than 70 years of age. Supraventricular tachycardias, of which atrial fibrillation was the most common, comprised 95.3% of the cases; bradyarrhythmia and ventricular ectopic beats were seen in only three and four cases, respectively. Arrhythmias were most likely to develop on the second day after the operation. Eighty percent of the arrhythmias disappeared within 3 days after onset, and sinus rhythm was finally restored with digitalis or other antiarrhythmic drugs in all patients except one, who had a myocardial infarction. Arrhythmias were not the direct cause of any of the seven in-hospital deaths. A multivariate analysis of 16 variables revealed that age and extent of pulmonary resection were significant risk factors. Despite these significant risk factors, arrhythmias after thoracic operations could be managed without special prophylaxis and were not closely related to higher mortality. PMID- 8246547 TI - Thoracoscopic pulmonary lobectomy. Early operative experience and preliminary clinical results. AB - Thoracoscopic video-assisted lobectomy procedures were performed in 11 patients (7 men, 4 women; age range 40 to 74 years, mean 66 years). Ten patients had peripheral pulmonary opacities: eight of these were bronchogenic carcinomas, one was an atypical carcinoid lesion, and one was a pulmonary infarct. All of these cases had preoperative evaluation by computed tomographic scanning to exclude mediastinal lymphadenopathy. The remaining patient had preoperatively diagnosed lobar bronchiectasis. Surgical access was gained via three stab (1 cm) incisions and one short (7 cm) submammary incision, which was made without rib separation and was used for specimen delivery. Lobes resected were the left upper (n = 4), left lower (n = 2), right upper (n = 2), and right lower (n = 3). All patients survived. Overall mean operative time was 3.3 hours and blood loss 263 ml. For the latter five cases, however, these figures were reduced to 2.3 hours and 100 ml, respectively, indicating improvement with experience. In no cases was ventilatory assistance required. Mean high-dependency unit time was 41 hours. In each case, it was possible to perform a standard dissectional lobectomy with lobar lymph node clearance equal to that obtained at open thoracotomy. Comparison with a series of 33 open lobectomy procedures demonstrated reduced postoperative pain, morphine consumption, and high-dependency unit stay. This preliminary experience supports the development of video-assisted thoracoscopic pulmonary lobectomy for patients with small peripheral opacities or known benign disease. PMID- 8246548 TI - Bronchoplastic procedures for tuberculous bronchial stenosis. AB - Thirty-six patients underwent tracheobronchoplastic procedures for treatment of tuberculous tracheobronchial stenosis. The modes of operations were left upper sleeve lobectomy in 13 patients, sleeve resection of the left main bronchus in 12 patients (two underwent concomitant left upper lobectomy), right upper sleeve lobectomy in five patients, sleeve resection of the right intermediate bronchus in two patients, right sleeve superior segmentectomy of the lower lobe in one patient, sleeve resection of the trachea with concomitant left pneumonectomy in one patient, carinal resection with right upper sleeve lobectomy and middle lobectomy in one patient, and dilatation of the left main bronchus with a free skin graft reinforced with a steel wire in one patient. One patient died of pulmonary edema of unknown cause on the first postoperative day. Anastomotic stenosis occurred in seven patients. One of these patients underwent reoperation and six underwent endoscopic dilatation. One patient died in the hospital of massive bleeding during endoscopic dilatation 4 months after operation. Slight to moderate stenosis resulted in the remaining patients. Although there are some complications, we believe bronchoplastic operation is worthwhile for restoring pulmonary function in patients with tuberculous tracheobronchial stenosis. PMID- 8246549 TI - Atrioventricular valve competence after takedown to improve exposure during ventricular septal defect repair. AB - Although the atrioventricular valve and its attachments can sometimes obscure the superior margin of a ventricular septal defect, concern for valvular competence has made surgeons hesitant to take down the atrioventricular valve. Over a 10 year period, the right atrioventricular valve was taken down to improve exposure for ventricular septal defect repair in 40 patients at our institution, and follow-up echocardiographic studies to determine the degree of valvular regurgitation were available in 32. On the basis of the area of the color flow jet, valvular regurgitation was graded as none in 22 and trivial in 10. Heart block did not develop in any patient, and there were no deaths. Takedown and resuspension of the atrioventricular valve is a safe and effective technique that improves exposure for ventricular septal defect repair and does not adversely affect valve competence. PMID- 8246550 TI - Specific sequelae after Fontan operation at mid- and long-term follow-up. Arrhythmia, liver dysfunction, and coagulation disorders. AB - Patients who have undergone a Fontan-type operation usually have an elevated systemic venous pressure. To determine the sequelae of this nonphysiologic condition, we evaluated 66 patients 1 to 14 years after a Fontan-type operation. Fifty-one patients were apparently in good clinical condition, and 15 patients had symptoms and were restricted in their daily life. Bicycle exercise capacity, tested in 41 patients, ranged from 50% to 110% (mean 85%) of the predicted value for length. In 16 patients, a decreased capacity (< 85%) was, among others, related to arrhythmias and the presence of protein-losing enteropathy. A 24-hour ambulatory electrocardiogram was available in 56 patients and found to be normal in 32 (57%) patients. Arrhythmias were present in 21 patients, six of whom had symptoms. Three patients had previous pacemaker implantation. One or more abnormalities in liver enzyme and function tests were present in 40 patients (61%) and in coagulation factors in 46 patients (69%). The most pronounced was a protein C deficiency, a known thrombotic risk factor, present in 41 patients. The occurrence of arrhythmias increased with time of follow-up (p < 0.004), the occurrence of protein C deficiency decreased with time (p < 0.0001), and the occurrence of abnormal liver enzyme and function tests was not related to time of follow-up. With regard to age at operation, arrhythmias did not occur in patients who underwent operation at a mean age of 4 +/- 1.9 years (standard deviation), in contrast to patients who underwent operation at a mean age of 7.6 +/- 4 years (standard deviation) (p < 0.001). The occurrence of the two other types of sequelae was not related to the age at operation. With regard to the type of operation, only patients with a valved right atrium-to-pulmonary artery connection had a higher prevalence of arrhythmias than patients with a nonvalved or direct right atrium-to-pulmonary artery connection (p < or = 0.001). The latter patients also had a higher prevalence of protein C deficiency (p < or = 0.001). No relationship was found among the other types of operation, the underlying structure, or the hemodynamic condition measured at rest and the presence of arrhythmias, abnormal liver enzyme and function tests, or protein C deficiency. This point survey shows that even patients with an apparently good clinical condition are at risk for arrhythmias, abnormal liver enzyme and function tests, and coagulation factor abnormalities. Serial statement of affairs is recommended to ensure that adequate preventive measures can be taken. PMID- 8246551 TI - Common arterial trunk, uncommon coronary arterial anatomy. AB - Macroscopic investigation was done in 44 postmortem specimens of hearts with common arterial trunk. In 38 hearts, the normal distribution in left and right coronary arteries was found. Of the coronary orifices, five were pinpoint and three showed a double orifice. The left coronary orifice was positioned in the posterior part of the truncus (p < 0.0001); the right coronary orifice was positioned in the right anterior and lateral part (p < 0.0001). In 19 hearts, coronary orifices were found above sinus level, left coronary orifices more often than right coronary orifices (p < 0.001). In seven hearts, type I truncus was found, in seven type II truncus was found, in 17 the truncus was intermediate between types I and II, in two type III truncus was found. In 11 hearts, the pulmonary artery distribution could no longer be identified. The truncal valve was bicuspid in 11 hearts, tricuspid in 25 hearts, and quadricuspid in eight hearts. The truncal valve showed overriding of 5% to 100%. Malformations of the coronary arteries were found in 28 hearts (64%). In 27 hearts (61%), the coronary arterial anatomy might have had clinical consequences. In nine hearts, coronary arterial orifices were at risk in excision of the pulmonary arteries from the common arterial trunk. The role of the neural crest as an etiologic factor of coronary arterial malformations in common arterial trunk should be taken into account. PMID- 8246552 TI - Effects of preserving mitral apparatus on ventricular systolic function in mitral valve operations in dogs. AB - The mitral apparatus can affect left ventricular function through various mechanisms, such as the direct mechanical coupling between the mitral anulus and papillary muscle and the mitral annular contraction. To evaluate the relative contribution of these mechanisms, we investigated in five groups of 35 dogs that underwent open chest operations how preservation of the mitral apparatus affects left ventricular systolic function. We performed atriotomy in the first group. We sutured a prosthetic rigid ring around the mitral anulus in the second group. We replaced the mitral valve and preserved the anterior chordae in the third group, the posterior chordae in the fourth group, and no chordae in the fifth group. The postoperative percentage of recovery of ventricular function (as assessed by the slope of the end-systolic pressure-volume relation) from preoperative control values were 92.2% +/- 4.8%, 89.5% +/- 12.8%, 85.7% +/- 9.5%, 75.1% +/- 12.9%, and 61.3% +/- 8.0%, respectively. Preservation of the mitral apparatus significantly improved left ventricular function compared with that of conventional mitral valve replacement. The average relative contribution of the ventricular muscle to left ventricular function, the mitral anulus-papillary muscle continuity, and the mitral annular contraction were 66.5%, 30.6%, and 2.9%, respectively. We conclude that it is more beneficial to preserve the mitral apparatus in mitral valve replacement. The prosthetic ring does not detract from the functional benefit of the preservation of the mitral apparatus. PMID- 8246553 TI - Structural basis for changes in left ventricular function and geometry because of chronic mitral regurgitation and after correction of volume overload. AB - Left ventricular function and myocyte structure were examined in three groups of dogs: (1) 3 months of mitral regurgitation caused by chordal rupture (n = 7); (2) chronic mitral regurgitation followed by mitral valve replacement and a 3-month recovery period (n = 7), and (3) sham controls (n = 8). The left ventricular end systolic stiffness constant (Kess) was measured as an index of left ventricular contractile function with stress-strain relationships obtained by cinecatheterization. Isolated myocyte structure and composition were examined with computer-assisted morphometry and nuclear area computed with deoxyribonucleic acid fluorescence. Left ventricular contractile function was significantly depressed with chronic mitral regurgitation compared with control values (Kess, 2.1 +/- 0.1 versus 3.6 +/- 0.2; p < 0.05) and returned to control values with mitral valve replacement (3.8 +/- 0.2). Left ventricular mass significantly increased in both the mitral regurgitation and mitral valve replacement groups compared with control values (121 +/- 10, 120 +/- 5 versus 95 +/- 9 gm, respectively; p < 0.05). Myocyte length increased with mitral regurgitation beyond control values (194 +/- 4 versus 218 +/- 8 microns; p < 0.05) and increased beyond mitral regurgitation values after mitral valve replacement (231 +/- 7 microns; p < 0.05). Myocyte volume with mitral regurgitation increased slightly beyond control values (33.5 +/- 0.7 versus 37.6 +/- 1.3 microns3; p = 0.15) and significantly increased with mitral valve replacement (40.1 +/- 1.2 microns3; p < 0.05). Myocyte myofibril volume significantly declined with mitral regurgitation compared with control values (14.8 +/- 1.5 versus 22.2 +/- 0.7 microns3; p < 0.05) and significantly increased beyond both mitral regurgitation and control values with mitral valve replacement (27.1 +/- 1.1 microns3; p < 0.05). Myocyte nuclear area with mitral regurgitation remained unchanged from control values (1430 +/- 122 versus 1163 +/- 89 microns2) but increased significantly with mitral valve replacement (2209 +/- 250 microns2; p < 0.05). In summary, the left ventricular contractile dysfunction with chronic mitral regurgitation is accompanied by increased myocyte length and reduced myofibril content. In contrast, the left ventricular hypertrophy and improved left ventricular pump function with mitral valve replacement were due to increased myocyte volume and increased contractile protein content. PMID- 8246554 TI - Mid-term follow-up after heart valve replacement with CarboMedics bileaflet prostheses. AB - The CarboMedics cardiac prosthesis (CarboMedics, Inc., Austin, Tex.) is a relatively new, low-profile bileaflet prosthesis. We report the results of a prospective mid-term evaluation of 426 CarboMedics prostheses implanted at our institution. Three hundred fifty patients had CarboMedics prostheses implanted in the mitral (n = 125), aortic (n = 149), or aortic and mitral positions (n = 76). Hospital mortality was 5.2%. Follow-up was 98% complete, with 478 patient/years and a mean follow-up of 19 months (range 1 to 42 months). Actuarial freedom from complications were calculated as follows (linearized rates in parentheses): late mortality 97% +/- 0.77% (2.7%), thromboembolism 98% +/- 0.4% (0.5%), anticoagulation-related hemorrhage 98.6% +/- 0.4% (0.4%), nonstructural dysfunction 98.5% +/- 0.6 (0.2%), valve-related mortality 91.5% +/- 0.7 (0.5%), valve failure 90.9% +/- 1% (0.7%), treatment failure 92.8% +/- 0.8% (0.7%), and all valve-related morbidity and mortality 90% +/- 2% (2.6%). According to our results, the CarboMedics valve has a low rate of complications that further improves the quality of life in patients with heart valve prostheses. PMID- 8246555 TI - Optimal shape of prosthesis for tricuspid valve replacement. AB - To develop an optimal prosthesis for use in the tricuspid valve, we determined the change in cardiac performance associated with the change in shape of the anulus of the tricuspid valve using a metal ring of variable flat ratio (defined as the long/short axis). In experiment I in an isolated autoperfusion model with six dogs, the left ventricular stroke work under a constant left atrial pressure of 7 mm Hg were, in grams per beat per minute, 11.88 +/- 0.52 with a flat ratio of 1.0, 11.88 +/- 0.70 with a flat ratio of 1.3, 16.23 +/- 0.61 with a flat ratio of 1.6, and 11.22 +/- 0.38 with a flat ratio of 2.3 (mean +/- standard error of the mean). The volume was significantly higher with a flat ratio of 1.6 than with the other ratios (p < 0.05). In experiment II with six dogs in vivo, the blood flow of the pulmonary trunk artery was 0.57 +/- 0.03 L/min with a flat ratio of 1.0, 0.66 +/- 0.03 L/min with a flat ratio of 1.6, and 0.58 +/- 0.03 L/min with a flat ratio of 2.3; the flow was significantly larger with a ratio of 1.6 than with the other ratios (p < 0.05). Right ventriculography was performed in five dogs. The percentage of radial shortening of the apex close to the acute margin was 19.4% +/- 3.1% with a flat ratio of 1.0 and 49.2% +/- 5.1% with a flat ratio of 1.6 (p < 0.05). In conclusion, the best hemodynamic results were obtained with a flat ratio of 1.6, which showed more advantageous hemodynamics than do the ratios of conventional circular prostheses. PMID- 8246556 TI - Cryopreserved aortic homografts contain viable smooth muscle cells capable of expressing transplantation antigens. AB - Frozen aortic tissue is increasingly used as homografts in reconstructive cardiovascular surgical procedures. The viability of cells within these cryopreserved tissues, their identity, and their potential immunogenicity have been the subject of controversy. We cultured cells from cryopreserved human aortic homografts that reacted with a monoclonal antibody that recognizes muscle actin isoforms, identifying them as smooth muscle cells. Under basal conditions, these smooth muscle cells contained messenger ribonucleic acid for class I human leukocyte antigens detected by northern blotting and expressed class I human leukocyte antigen on their surfaces as measured by enzyme-linked immunoassay and immunohistochemistry. Unstimulated smooth muscle cells contained no class II human leukocyte antigen messenger ribonucleic acid as determined by northern blotting and displayed almost no class II surface antigen as determined by enzyme linked immunoassay. Interferon gamma (1000 U/ml, 72 hours), a product of activated T lymphocytes, not only increased the expression of class I human leukocyte antigens by smooth muscle cells, but induced class II human leukocyte antigen messenger ribonucleic acid and elevated surface expression from 22 +/- 7 to 819 +/- 35 enzyme-linked immunoassay units (n = 4). Immunohistochemistry revealed few class II-positive smooth muscle cells under basal culture conditions, but all cells showed high levels of DR antigen after exposure to interferon gamma for 3 days. Similar results were obtained in two independent isolates. We conclude that cryopreserved aortic homografts can contain viable smooth muscle cells capable of expressing major histocompatibility antigens that might render them immunogenic and susceptible to rejection by the recipient's immune system. PMID- 8246557 TI - Frame-mounted porcine valve bioprostheses: preparation during aortic-root dilation. Biomechanics and design considerations. AB - The leaflet geometry and function of frame-mounted porcine bioprostheses prepared with dilation of the aortic root during frame mounting was investigated. The diameter of the porcine aortic root increased by 47% when dilated with a pressure of 120 mm Hg. In the absence of pressure dilation, the ratio of circumferential leaflet length to the radius of the aortic root (S/RA) was 2.7 +/- 0.1, and the angle of inclination of the leaflet to the base of the valve was 17 +/- 4.3 degrees. In this condition, the leaflet geometry was similar to that of some second-generation porcine bioprostheses, which demonstrated high open-leaflet bending strains at the commissures. Dilation of the porcine root with 120 mm Hg reduced the value of S/RA to 1.84 and produced a triangular open-leaflet configuration with minimum open-leaflet bending strains. Open-leaflet bending strains were also reduced in two prototype frame-mounted valves prepared with partial dilation of the aortic root, which had an S/RA ratio of less than 2.3 and a leaflet angle greater than 27 degrees. The study indicates that the next generation of porcine bioprostheses should be prepared with at least 17% dilation of the aortic root during frame mounting and with zero pressure difference across the leaflets during fixation. This can be achieved by applying an equal hydrostatic pressure to both sides of the valve leaflets during fixation and frame mounting. PMID- 8246558 TI - Surgical treatment of postinfarction false aneurysm of the left ventricle. AB - Twelve patients underwent surgery for repair of false aneurysms of the left ventricle. The mean interval between the myocardial infarction and the diagnosis was 19 months (range 2 to 80 months). Congestive heart failure was the most common clinical presentation. Most patients had three-vessel coronary artery disease. The false aneurysm was posterior in 10 patients and anterior in two. Three patients with posterior aneurysm had severe mitral regurgitation. Repair was accomplished by resection of the false aneurysm and primary closure of the defect in four patients and by closure with a patch in eight. Nine patients also had coronary artery bypass. Mitral valve replacement was performed in three patients who had severe mitral regurgitation before the operation and in one patient who had severe mitral regurgitation after repair of the false aneurysm and could not be weaned from cardiopulmonary bypass. There were three operative deaths and one additional death after 2 months. All deaths occurred in patients who had mitral valve replacement. Eight patients survived the operation and remained well after a mean follow-up period of 62 months. Patients with false aneurysms of the left ventricle do well after surgical repair, except when concomitant mitral valve replacement is necessary. PMID- 8246559 TI - Internal thoracic artery: prevalence of atherosclerotic changes. AB - Use of the internal thoracic artery for myocardial revascularization has regained general acceptance because it offers better long-term results than do venous conduits. The aim of this study was to ascertain the prevalence of atherosclerosis in the internal thoracic artery and to correlate the prevalence with other known risk factors. A total of 117 patients (male/female ratio 84:33; mean age 56.8 years) were investigated. Sixty-eight patients had coronary artery disease, 25 had combined coronary artery and valvular heart disease, 14 had acquired valvular heart disease, and 10 had other types of heart disease. All but one patient underwent bilateral semiselective internal thoracic arteriography. Evidence of atherosclerotic change was present in 6.6% of the opacified vessels in 11.1% of the investigated individuals. Although all patients with atherosclerotic lesions in the internal thoracic artery had coronary artery disease, no correlation could be found between coronary artery disease and internal thoracic atherosclerosis. Peripheral vascular disease and hyperlipidemia could be identified as predictors of atherosclerotic changes in the internal thoracic artery. Atherosclerosis is somewhat more prevalent in the internal thoracic artery in this study than in the literature. Although the internal thoracic artery is a protected vessel, there is a certain extent of atherosclerosis, that correlates with known risk factors. Our observations should not preclude use of the internal thoracic artery, but they should be considered for patients who are at risk for atherosclerotic changes of the internal thoracic artery. PMID- 8246560 TI - Transplantation of hearts after arrest and resuscitation. Early and long-term results. AB - Transplant surgeons are reluctant to use hearts that have undergone cardiopulmonary resuscitation for cardiac arrest because of the fear of poor early and late cardiac function. A policy of minimizing contraindications to use of donor hearts has led to the unique opportunity of assessing the effects of donor arrest and successful cardiopulmonary resuscitation on early and late cardiac function in pediatric heart transplantation. A number of 140 infants and children undergoing transplantation from birth to 17 years of age were studied retrospectively and divided into two groups on the basis of cardiopulmonary resuscitation status. Group 1 (72 patients) received donor hearts that were not subjected to cardiopulmonary resuscitation; group 2 (68 patients) received donor hearts that had cardiopulmonary resuscitation for a mean of 18.8 +/- 14.6 minutes, the longest period of time being 60 minutes. Mean ischemic times were almost identical in the two groups: 4.43 +/- 2.0 hours (cardiopulmonary resuscitation) versus 4.5 +/- 2.1 hours (no cardiopulmonary resuscitation). Early cardiac function was assessed on the basis of the number of days the recipient was supported by the ventilator, days receiving dopamine, days receiving isoproterenol, and the amount of inotropic agents required after the operation. The groups did not differ. Parameters of systolic function included fractional shortening, posterior wall thickening, and maximum velocity of change in left ventricular posterior wall dimension during systole. Diastolic function was measured on the basis of left ventricular end-diastolic volume, left ventricular mass, and maximum velocity of change in left ventricular posterior wall dimension during diastole. Both systolic and diastolic function were measured and analyzed from M-mode echocardiography at 1 week, 1 month, 6 months, 1 year, and 2 years after the operation. There were no statistically significant differences in graft function between the two groups in any of the echocardiographic parameters studied, even at 2 years. No group differed from ranges of normal. Our results suggest that hearts undergoing cardiopulmonary resuscitation for periods of up to 60 minutes can be used safely without evidence of deterioration of early or late cardiac function. PMID- 8246561 TI - Prospective evaluation of malignant neoplasms in cardiac transplant recipients uniformly treated with prophylactic antilymphocyte globulin. AB - Although there is convincing evidence that prophylactic administration of high doses of the monoclonal antibody OKT3 predisposes patients to an increased prevalence of early posttransplantation malignancy, particularly posttransplantation lymphoproliferative disease, it is indeterminate whether polyclonal antilymphocyte globulin poses a similar hazard. We reviewed the outcome of 112 consecutive cardiac transplant recipients who received uniform immunosuppression, including induction therapy with antilymphocyte globulin, and were prospectively followed-up for a median duration of 41.5 months (range 1 to 81 months). No patients had posttransplantation lymphoproliferative disease. Nine malignant neoplasms (8%) were detected from 6 to 70 months after transplantation. Four patients with cutaneous neoplasms were alive and well at the time this article was written. Three patients died of disseminated adenocarcinoma 6 months, 17 months, and 60 months after transplantation. One patient was undergoing treatment of Kaposi's sarcoma at the time this article was written, and another was undergoing treatment of transitional bladder cell carcinoma. Actuarial survival for all patients was 88% at 1 year and 79% at 5 years. Moderate doses of induction antilymphocyte globulin may facilitate rapid reduction of maintenance cyclosporine and steroid doses, thereby decreasing the duration of intense immunosuppression and lowering the risk of posttransplantation lymphoproliferative disease. Testing this hypothesis would require the development of reliable and reproducible in vivo assays to prospectively assess immune status. PMID- 8246562 TI - Cardiac fibroma and heart transplantation. AB - Successful orthotopic heart transplantation was performed in a 38-day-old child with a fetal echocardiographic diagnosis of a left ventricular mass and in a 40 year-old woman with cardiac murmur since childhood and an echocardiographic diagnosis of asymmetric septal hypertrophy. Pathologic examination of the removed hearts, consisting of gross, histologic, immunohistochemical, and ultrastructural studies, led to the final diagnosis of cardiac fibroma. Both patients were alive and in good condition at 35 and 28 months, respectively, after operation. PMID- 8246563 TI - Cold ischemia and reperfusion each produce pulmonary vasomotor dysfunction in the transplanted lung. AB - Pulmonary vascular resistance is significantly increased in the transplanted lung. We hypothesized that the ischemic or reperfusion injuries incurred by the transplanted lung may produce pulmonary vasomotor dysfunction, which in turn may produce increased pulmonary vascular resistance. In a dog model of autologous lung transplantation, the purpose of this study was to examine the following mechanisms of pulmonary vasomotor control and to relate each of them to cold ischemia and to reperfusion: (1) endothelium-dependent cyclic guanosine monophosphate-mediated vasorelaxation (response to acetylcholine 10(-6) mol/L), (2) endothelium-independent cyclic guanosine monophosphate-mediated vasorelaxation (response to sodium nitroprusside 10(-6) mol/L), and beta adrenergic cyclic adenosine monophosphate-mediated vasorelaxation (response to isoproterenol 10(-6) mol/L). Autologous right lung transplantation was performed in five dogs. At each of three times, two third-order pulmonary arteries were dissected from each transplanted lung and studied: control (immediately after harvest), cold ischemia (3 hours in 4 degrees C saline solution), and cold ischemia plus reperfusion (1 hour after lung reimplantation). The vasorelaxing effects of acetylcholine, sodium nitroprusside, and isoproterenol were studied in isolated pulmonary arterial rings, suspended on fine wire tensiometers in individual organ chambers. Statistical analysis was by analysis of variance. Results demonstrated significant dysfunction of beta-adrenergic cyclic adenosine monophosphate-mediated relaxation after cold ischemia alone, and this dysfunction was exacerbated by reperfusion. Endothelium-dependent cyclic guanosine monophosphate-mediated relaxation was not impaired by cold ischemia alone but was significantly impaired by reperfusion. Endothelium-independent cyclic guanosine monophosphate-mediated relaxation was not impaired by cold ischemia or reperfusion. We conclude that cold ischemia and reperfusion each produce different patterns of pulmonary vasomotor dysfunction. Cumulatively, such dysfunction may contribute to increased pulmonary vascular resistance in the transplanted lung. PMID- 8246564 TI - Medtronic intact bioprosthesis in aortic position: unexplainable immediate degeneration. PMID- 8246565 TI - Isolated anomalous origin of left pulmonary artery from the ascending aorta in an adult. PMID- 8246566 TI - Bidirectional cavopulmonary anastomosis. PMID- 8246567 TI - Left ventricular outflow tract obstruction and mitral regurgitation caused by papillary muscle abnormalities: a case report. PMID- 8246568 TI - Artificial chordal reconstruction with expanded polytetrafluoroethylene sutures for mitral valve replacement in a patient with mitral stenosis: case report. PMID- 8246569 TI - A self-retaining retractor for operations for congenital heart disease. PMID- 8246570 TI - Histoacryl tissue adhesive: an alternative means of stopping polytetrafluoroethylene graft sweating. PMID- 8246571 TI - Cystic fibroelastoma of the mitral valve: report of a case. PMID- 8246572 TI - Migrated Kimray-Greenfield filter lodged in the right ventricle: surgical removal and transatrial reinsertion. PMID- 8246573 TI - Aortopulmonary paraganglioma, a rare aortic tumor: a case report. PMID- 8246574 TI - Unusual malignancy in the wall of a mediastinal cyst. PMID- 8246575 TI - Coexisting Wolff-Parkinson-White syndrome and cardiac valve disease: a staged treatment approach with catheter ablation before operation. PMID- 8246576 TI - Use of the heart from the older donor for the older transplant recipient. PMID- 8246577 TI - Desmopressin does not decrease bleeding after cardiac operation in young children. AB - Young children undergoing complex cardiac operation lose more blood after cardiopulmonary bypass than do older patients. This study was designed to investigate the effect of desmopressin on blood loss during the first 24 hours after cardiac operation in children undergoing principally complex surgical procedures. The study consisted of a randomized, blinded comparison of 112 pediatric patients who received either desmopressin 0.3 microgram/kg or saline solution placebo after cardiopulmonary bypass. A coagulation profile including bleeding time, quantitation of von Willebrand factor, and qualitative analysis of the factor VII:von Willebrand factor complex was performed before, 30 minutes after, and 3 hours after the operation. Blood loss and blood replacement were recorded for the first 24 hours after the operation. The surgeon classified the technical difficulty of each procedure as simple or complex. Statistical analysis was performed with Student's unpaired t test and chi 2 analysis. Significance was defined as p < 0.05. Results are listed as mean +/- standard deviation. Data collection was completed for 95 patients. The mean age of all patients was 26 +/- 40 months, and the mean weight was 10 +/- 11 kg, with 84% undergoing complex procedures. There were no differences between the desmopressin and placebo groups with respect to age, weight, or surgical complexity. Twenty-four-hour blood loss and replacement between the desmopressin and placebo groups were not different (blood loss: desmopressin 30 +/- 33 ml/kg, placebo 35 +/- 36; blood replacement: desmopressin 65 +/- 43 ml/kg, placebo 64 +/- 46 ml/kg). Coagulation profiles between the desmopressin and placebo groups were not different at any time. We conclude that desmopressin does not reduce blood loss or blood replacement in young children after cardiopulmonary bypass for either simple or complex cardiac surgical procedures. PMID- 8246578 TI - Thromboxane A2 mediates pulmonary hypertension after cardiopulmonary bypass in the rabbit. AB - Clinically, it is well established that cardiopulmonary bypass results in pulmonary dysfunction. Using a recently developed preparation for cardiopulmonary bypass in the rabbit, we have been able to mimic a similar, but more severe, condition. We found that, despite normal histologic structure of the myocardium, hearts could not be weaned from bypass because of a serious increase in pulmonary vascular resistance. Histologic studies of the lungs showed severe intravascular neutrophil aggregation and marked vasoconstriction. To identify the nature and origin of the mediator responsible for the changes in the pulmonary vasculature, we subjected groups of rabbits (n = 4 per group) to bypass with cooling to 18 degrees C, circulatory arrest for 1 hour, and rewarming on bypass to 33 degrees C. Pulmonary vascular resistance was measured at the same temperature before and after bypass. Four groups were studied: group I were untreated controls; group II received the cyclooxygenase inhibitor, indomethacin (0.2 mg/kg intravenously), before operation; group III received the thromboxane A2 synthetase inhibitor, Dazmegral (5 mg/kg intravenously), before operation together with the thromboxane A2 receptor blocker GR 32191B (2 mg/kg per 30 minutes intravenously); and group IV were treated with mustine hydrochloride (1.75 mg/kg intravenously) 3 days before the experiment to deplete the neutrophils by 90%. During circulatory arrest, the heart was protected with an initial infusion (10 ml at 4 degrees C over 1 minute) of St. Thomas' Hospital cardioplegic solution. At the end of the experiment, the heart and lungs were histologically examined. In the control group, a significant increase (+395% when compared with the value recorded before bypass) in pulmonary vascular resistance was observed after bypass. However, in none of the treated groups did pulmonary vascular resistance increase significantly (percentage changes in groups II, III, and IV were -24%, 0%, and +33%, respectively). Pulmonary histologic characteristics were normal in all treated groups, and all animals were successfully weaned from bypass. These results indicate that the increase in pulmonary vascular resistance that arises as a consequence of bypass in rabbits is primarily a result of the production of thromboxane A2, a process in which the neutrophil plays a pivotal role. PMID- 8246579 TI - Cardiopulmonary bypass significantly reduces surfactant activity in children. AB - Lung injury remains an important problem after cardiopulmonary bypass. The contribution of altered surfactant concentration or activity to pulmonary dysfunction after cardiopulmonary bypass is unclear. Recent evidence indicates that alveolar surfactant exists in specific aggregate forms that differ with respect to density, phospholipid composition, and function. A transition from surface active, higher density, large aggregates of surfactant to lower density, small aggregates that possess reduced surface activity has been demonstrated after experimental lung injury. The purpose of the present study was to examine surfactant aggregate fractions before and after bypass in children. Twelve acyanotic patients, aged 2 to 12 years, underwent intraoperative pulmonary function testing followed by bronchoalveolar lavage before incision and approximately 1 hour after termination of cardiopulmonary bypass. Saturated phosphatidylcholine pool sizes and total protein content of the small- and large aggregate fractions of bronchoalveolar lavage fluid were determined. One hour after termination of cardiopulmonary bypass, the ratio of saturated phosphatidylcholine in small-aggregate as compared with that in large-aggregate fractions increased (mean +/- standard error) from 0.19 +/- 0.03 to 0.37 +/- 0.07 (p < 0.02), as did the ratio of saturated phosphatidylcholine to protein in the small-aggregate fraction (from 0.04 +/- 0.01 to 0.08 +/- 0.02, p < 0.05). Reductions in forced vital capacity (-19% +/- 5%), inspiratory capacity (-15% +/- 3%), and small airway flow rates (-32% +/- 6%) were also observed after bypass. These changes were accompanied by a fivefold increase in alveolar polymorphonuclear leukocyte content. The present study suggests that cardiopulmonary bypass of moderate duration in relatively healthy children is associated with surfactant changes that are similar in type and magnitude to those observed in experimental lung injury. PMID- 8246580 TI - Complement activation during cardiopulmonary bypass in infants and children. Relation to postoperative multiple system organ failure. AB - Twenty-nine children 3 months to 17 years of age undergoing operations for congenital heart disease were included in this prospective study. Complement activation, activation of the plasma contact system, leukocytes, leukocyte elastase release, and C-reactive protein were studied during and after cardiopulmonary bypass for the first postoperative week and related to multiple system organ failure occurring in eight (27.5%) of the 29 children. During cardiopulmonary bypass complement activation via the alternative pathway as indicated by significant conversion of C3 (expressed by C3d/C3) and abnormally high C5a values at the end of cardiopulmonary bypass without consumption of C4 was shown in all children. At the end of cardiopulmonary bypass, C3 conversion was significantly higher in the eight patients with multiple system organ failure than in the others (p < 0.05), whereas no difference in C5a level was shown. All children had a significant increase in leukocyte count directly after protamine administration (p < 0.0001) and elastase release during cardiopulmonary bypass that was significantly higher in patients with multiple system organ failure than in those without (p < 0.05). Consumption of prekallikrein as an indicator of activation of the Hageman system was not detectable during cardiopulmonary bypass in any child. After cardiopulmonary bypass, in patients without multiple system organ failure, C3d/C3 decreased and reached preoperative values within the first postoperative week, whereas, in patients with multiple system organ failure, C3d/C3 increased further, reaching a maximal value on the third postoperative day. In comparison with patients without multiple system organ failure, patients with multiple system organ failure showed a severe decrease of C4 (with minimal values on the third postoperative day), suggesting consumption by activation of the classic pathway of the complement system or a hepatic synthesis deficiency. Prekallikrein values were also significantly lower in patients with multiple system organ failure than in the others, with a maximal difference on the third postoperative day (p < 0.005). C-reactive protein was significantly lower in patients with multiple system organ failure than in the others for the first 2 postoperative days (p < 0.05), probably because of severe hepatic failure in patients with multiple system organ failure. This study demonstrates that, in children, cardiopulmonary bypass induces complement activation principally via the alternative pathway. It suggests a relationship between complement activation and multiple system organ failure observed in the postoperative period. Furthermore, it points out the role of multiple system organ failure itself on the C3 conversion and on the synthesis of the markers of the inflammatory response in children after heart operations. PMID- 8246581 TI - Right ventricular function after normothermic versus hypothermic cardiopulmonary bypass. AB - Normothermic systemic perfusion in patients undergoing cardiopulmonary bypass may compromise myocardial hypothermia, a mainstay for preservation of ventricular function during iatrogenic cardiac arrest. The right ventricle is the area of the heart most susceptible to rewarming. We prospectively evaluated myocardial rewarming and indexes of right ventricular function in 30 patients undergoing coronary artery bypass grafting randomized to receive moderate hypothermic (bladder temperature 25 degrees C) or normothermic perfusion and multidose cold blood cardioplegia during cardiopulmonary bypass. All patients had significant stenosis (> 70%) of the right coronary artery, and in 27 of 30 the right coronary artery was revascularized. A right ventricular ejection fraction/volumetric catheter was used to assess right ventricular function by right ventricular ejection fraction and a preload (right ventricular end-diastolic volume) normalized right ventricular stoke work index in the prebypass and postbypass periods. Findings included the following: (1) Greater rewarming of all areas of the heart occurs with normothermic bypass, with the mean temperature difference at the end of each intracardioplegic period ranging from 4.0 degrees to 6.3 degrees C warmer than with hypothermic bypass; (2) the right ventricle was not more susceptible to rewarming than the posterior left ventricle or interventricular septum in either group; (3) right ventricular function did not differ between groups at any time in the study, including the immediate postarrest period; and (4) right ventricular function was preserved and equivalent to the prebypass baseline. We conclude that the moderate myocardial rewarming that occurs with normothermic perfusion does not compromise right ventricular preservation in patients with right coronary artery disease undergoing revascularization with multidose cold blood cardioplegia to maintain electromechanical arrest. PMID- 8246582 TI - The effects of pressure and flow on hemolysis caused by Bio-Medicus centrifugal pumps and roller pumps. Guidelines for choosing a blood pump. AB - Two Bio-Medicus BP-50 centrifugal pumps and two roller pumps were tested simultaneously with porcine blood at 21 degrees +/- 1 degree C in four in vitro circuits to determine the effect of four combinations of flow and pressure conditions on blood damage. Flows of 300 ml/min (1/4-inch inner-diameter tubing in the roller pump) and 1775 ml/min (1/2-inch inner-diameter tubing in the roller pump) and pressure differences across the pump (delta P = outlet pressure--inlet pressure) of 215 mm Hg (n = 6) and 345 mm Hg (n = 5) were examined. The index of hemolysis (milligrams plasma hemoglobin per 100 L blood pumped) for the BP-50 pump was higher at a flow of 300 ml/min than at a flow of 1775 ml/min (p < 0.0002). At 300 ml/min, the index of hemolysis for the BP-50 pump tended to be higher at 345 mm Hg than at 215 mm Hg (mean +/- standard error of the mean, 135 +/- 22 versus 88 +/- 9, p = 0.059). At 1775 ml/min, there was no difference in the index of hemolysis for the BP-50 pump between 215 and 345 mm Hg (37 +/- 7 versus 29 +/- 5, p = 0.32). With the roller pump, the index of hemolysis was higher at a flow of 300 ml/min than at a flow of 1775 ml/min (p < 0.036), but there was no difference in the indexes of hemolysis between 215 and 345 mm Hg at 300 ml/min (60 +/- 9 versus 61 +/- 11, p = 0.93) or at 1775 ml/min (40 +/- 6 versus 36 +/- 6, p = 0.61). Comparison between the two types of pumps showed that the index of hemolysis was significantly higher for the BP-50 than for the roller pump at a flow of 300 ml/min and a delta P of 215 mm Hg (88 +/- 9 versus 60 +/- 9, p = 0.009), as well as at a flow of 300 ml/min and a delta P of 345 mm Hg (135 +/- 22 versus 61 +/- 11, p = 0.001). At a flow of 1775 ml/min, there was no difference in the index of hemolysis between the two pumps at either pressure condition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246583 TI - Clinical observation on treatment of bradyarrhythmia with Chinese herbal medicine. AB - This paper reports on treatment of 62 cases of bradyarrhythmia with Fu Ben Zeng Mai Tang, a decoction of herbal medicine which has the effect of warming and tonifying the heart and kidney, reinforcing qi and improving blood circulation. The result indicates that it is effective for quickening the heart rhythm, improving the symptoms and regulating the function of sinoatrial node. It is also good for regulating the function of endocrine and the immune and nervous system. This therapy was compared with Western medicine in the treatment of 24 cases. PMID- 8246584 TI - Effect of acupuncture on weight loss evaluated by adrenal function. AB - In order to understand the relationship between patients' adrenal function and simple obesity and effect of acupuncture on it, the obesity indices, lipid indices, fasting blood-glucose, noradrenaline, dopamine, adrenalin and cortisol were observed. The results indicated that patients with simple obesity had hypofunction of the sympathetic-adrenal system and the hypothalamus-pituitary adrenal system. Acupuncture treatment not only affected weight loss but also enhanced functioning of the two systems, suggesting that the effect of acupuncture on weight loss may be produced by enhancing the functions of both the sympathetic-adrenal system and the hypothalamus-pituitary-adrenal system. PMID- 8246585 TI - Effect of acupuncture and point-injection treatment on immunologic function in rheumatoid arthritis. AB - The results of treatment of 54 cases of rheumatoid arthritis (RA) by warm needling (WN) and point-injection (PI) with Zhuifengsu are reported. Good clinical results were observed with an effective rate of 100%. At the same time, changes in cellular and humoral immunity and other parameters in peripheral blood were noted before and after treatment. The NK activity and IL-2 value in RA patients were found to be lower than those of normal individuals; both increased after treatment (P < 0.01). This suggests that the WN and PI with Zhuifengsu exert a regulatory effect on the cellular immunological function. PMID- 8246586 TI - The clinical application of yanglingquan (GB 34) point. PMID- 8246587 TI - Treatment of apoplectic hemiplegia with scalp acupuncture in relation to CT findings. PMID- 8246588 TI - 47 cases of acne treated by prick-bloodletting plus cupping. PMID- 8246589 TI - Treatment of adolescent acne with acupuncture. PMID- 8246590 TI - Effects of acupuncture on myoelectric activity of Oddi's sphincter in humans. PMID- 8246591 TI - Treatment of 60 cases of dysfunction of temporomandibular joint by puncturing zusanli (St 36) acupoint. PMID- 8246592 TI - Bloodletting at weizhong point (UB 40) for treatment of acute lumbar sprain. PMID- 8246593 TI - 75 cases of simple obesity treated with auricular and body acupuncture. PMID- 8246594 TI - 268 cases of myopia treated with injection and pellet pressure at auriculoacupoints. PMID- 8246595 TI - Treatment of periomarthritis with scalp acupuncture therapy--a report of 210 cases. PMID- 8246596 TI - Treatment of hiccough with acupuncture on middle sifeng. PMID- 8246597 TI - Treatment of facial spasm by plum-blossom needle tapping and cupping therapy. PMID- 8246598 TI - Clinical application of acupoint renying (St 9). PMID- 8246599 TI - An experimental study on acupuncture anti-hemorrhagic shock. AB - The effect of acupuncture at Neiguan (P 6) on regulating hemorrhagic shock in 90 rabbits was observed. All the results suggested that acupuncture at Neiguan could raise blood pressure, protect cardiac pump function, correct the disturbance of secretion and metabolism of humoral factors, reduce blood adhesion to normal level and thus achieve anti-hemorrhagic shock. This study has provided scientific basis for expounding the mechanism of acupuncture in anti-shock. PMID- 8246600 TI - A clinical and experimental study of herbal medicine in aplastic anemia. PMID- 8246601 TI - An introduction to keeping-fit massage. PMID- 8246602 TI - Treatment of xenophonia in male youths by extralaryngeal massage and language training. PMID- 8246603 TI - Recent advances in studies on traditional Chinese anti-aging materia medica. AB - Presented in this paper is a report of our studies on 386 traditional effective anti-aging medications, the effects of which on cell generation, survival time, immunomodulation, improvement of visceral and metabolic functions, and anti infection, and their trace element contents were further summarized and analysed. This suggests that the investigations of traditional anti-aging materia medica in China are now well under way and some effective drugs and compound prescriptions have been explored, such as Ginseng, Radix Astragali seu Hedysari, Radix Angelicae Sinensis, Herba Epimedii, Cordyceps, Ganoderma Lucidum seu Japonicum, Radix Polygoni Multiflori, Radix Acanthopanacis Senticosi, Rhizoma Polygonati, Fructus Lycii, and Poria. However, all of these preliminary results remain to be further investigated. PMID- 8246604 TI - Keeping-fit moxibustion (6). PMID- 8246605 TI - Treatment of headache with traditional auriculoacupoint therapy. PMID- 8246606 TI - How is impotence treated with acupuncture? PMID- 8246607 TI - The effects of GM-CSF on myeloperoxidase release in normal and myelodysplastic neutrophils. AB - When purified control neutrophils were primed with GM-CSF, a significant increase in FMLP-induced MPO release was observed (mean +/- S.E.M., 3.4 +/- 0.8 mU/10(7) unprimed cells compared to 6.5 +/- 1.1 mU/10(7) primed cells, p < 0.001). This MPO release was greatly augmented by Cytochalasin B (Cy B), but after the addition of Cy B the priming effects of GM-CSF became less obvious. Exposure to GM-CSF without FMLP did not enhance MPO release. Within whole blood, FMLP produced negligible MPO release, but priming with GM-CSF prior to FMLP always resulted in a significant increase in MPO release. Myelodysplastic neutrophils released similar amounts of MPO in response to FMLP, compared with control cells (3.4 +/- 0.8 mU/10(7) control cells compared to 2.7 +/- 0.3 mU/10(7) MDS cells, p > 0.05). Priming with GM-CSF produced an increase in FMLP-stimulated MPO release comparable with control cells. In terms of total MPO content, although some MDS patients exhibited low levels, as a group there was no significant difference from controls (169 +/- 21 mU/10(7) control cells compared with 157 +/- 19 mU/10(7) MDS cells). These findings suggest that MPO activity is not a universal defect in MDS and cannot account for the defects in respiratory burst activity in these neutrophils. PMID- 8246608 TI - Mutational inactivation of the p53 gene in the human erythroid leukemic K562 cell line. AB - The K562 human chronic myelogenous leukemia (CML) cell line has attained widespread use as a model for studying hematologic malignancy and erythroid differentiation. Sequencing of the p53 gene in the K562 cell line demonstrated a mutation in exon 5 characterized by a single base insertion (cytosine) between codons 135 and 136. This frameshift mutation leads to an N-terminal truncated protein of 147 amino acids. Only the mutated sequence was present suggesting that the normal allele has been lost. Reverse transcription PCR (RT-PCR) detected a p53 transcript but Western blotting and immunohistochemical staining of cells failed to detect p53 protein. The identification of an inactivation mutation of p53 in the K562 cell line further supports the argument that p53 mutations play a role in myeloid blast transformation of CML. PMID- 8246609 TI - Membrane-associated procollagenase of leukemic cells is activated by urokinase type plasminogen activator. AB - Murine leukemic cells from the WEHI-3B line, present in the cell surface a latent collagenase activity which is activated proteolytically. In this paper we show that this enzyme is activated by plasmin generated by the activity of a urokinase like plasminogen activator (u-PA) also present in the surface of these cells. Using a reverse fibrin autography method we found that u-PA is the major proteolytic activity present in the cell membranes. This fact suggests that u-PA could represent a normal activating system for this collagenase. PMID- 8246610 TI - Constitutive production of interleukin-8 (IL-8) by normal and malignant human B cells and other cell types. AB - The culture supernatants from 43 human cell lines obtained during log phase and from purified normal peripheral blood B-lymphocytes cultured at 10(6) cells ml-1 for 48 h in RPMI 1640-5% fetal calf serum were examined for interleukin-8 (IL-8) using Elisa kits. Constitutive IL-8 production was found for 14/15 B-cell lines (5 derived from normal persons and 2 from AML patients, 1 pre-B-ALL, 2 CLL with trisomy 12, 2 HTLV-I+, 1 HTLV-II+, 1/2 Burkitt lymphoma), 4/16 T-cell lines (3/6 HTLV-I+, 1 HTLV-II+, 0/9 T-ALL), myeloid line HL-60, monocytoid line U937, 3/3 ovarian carcinoma, 1/1 endometriosis, 2/2 normal fibroblast, 0/2 C-ALL, 0/1 pre erythroid line K562, as well as for normal B-lymphocytes. Later, cells examined by indirect immunofluorescence using IL-8 antibodies gave a positive reaction. DNA from 4 IL-8 producing and 3 non-producing cell lines, when probed with IL-8 cDNA gave the same 3.5 kb EcoRI fragment indicating similarities of the IL-8 gene in these cells. Two B-cell lines examined showed the expression of 1.8 kb IL-8 mRNA. These results indicate IL-8 production by a greater variety of cells than previously believed which open possibilities for new IL-8-mediated immune functions by such cells as B-cells. PMID- 8246611 TI - Are we curing acute myelogenous leukemia? PMID- 8246612 TI - Molecular diagnosis and monitoring of leukemia and lymphoma. PMID- 8246613 TI - 75th anniversary of Mayo dietetics. PMID- 8246614 TI - The incidence and prevalence of congestive heart failure in Rochester, Minnesota. AB - Although congestive heart failure is a fairly common clinical syndrome and the societal costs associated with its care are high, relatively little is known about the incidence or prevalence of the condition in the community. Using the resources of the Rochester Epidemiology Project, we identified all 46 persons 0 through 74 years of age who had a new diagnosis of congestive heart failure during 1981 and all 113 persons with a prevalent diagnosis on Jan. 1, 1982, in the city of Rochester, Minnesota. After confirming the diagnosis in the medical record by using criteria similar to those in the Framingham study, we found the annual incidence of congestive heart failure to be 110 per 100,000 after adjusting for age. Incidence rates were higher among male than among female study subjects (157 versus 71 per 100,000). In both male and female subjects, the incidence generally increased with advancing age, reaching 1,618 per 100,000 and 981 per 100,000, respectively. Prevalence rates on Jan. 1, 1982, demonstrated similar patterns. Overall, the prevalence of congestive heart failure was higher among male than among female subjects (327 versus 214 per 100,000) and increased exponentially with advancing age, reaching almost 3% in both sexes. Survival after a diagnosis of congestive heart failure was extremely poor, with only 80% alive at 3 months and 66% at 1 year. These data underscore the effect of congestive heart failure in the community and provide estimates of the number of persons who might benefit from early intervention. PMID- 8246615 TI - Rhinoscleroma: a growing concern in the United States? Mayo Clinic experience. AB - Rhinoscleroma is a chronic, progressive, granulomatous infection of the upper airways caused by the bacterium Klebsiella rhinoscleromatis. Although most cases occur in developing countries, recent immigration patterns have led to an increasing number of patients with rhinoscleroma in the United States. Rhinoscleroma may mimic various inflammatory and neoplastic processes, including leprosy, paracoccidioidomycosis, sarcoidosis, basal cell carcinoma, and Wegener's granulomatosis. Current therapy consists of a combination of surgical debridement and prolonged antimicrobial therapy. Rhinoscleroma should be added to the list of opportunistic infections that can occur in patients with human immunodeficiency virus. PMID- 8246616 TI - Transient constrictive pericarditis: diagnosis by two-dimensional Doppler echocardiography. AB - Patients with constrictive pericarditis usually require pericardiectomy to relieve their symptoms. In some patients, however, constrictive pericarditis may resolve spontaneously or with medical treatment. Four patients with transient constrictive pericarditis are described in this report. Although the cause of pericarditis differed, all patients had a small to large amount of pericardial effusion, followed by symptoms, signs, and Doppler features typical of constrictive pericarditis. Symptomatic improvement occurred after treatment with some combination of nonsteroidal anti-inflammatory agents, corticosteroids, and antibiotics. The resolution of the symptoms paralleled the normalization of characteristic respiratory changes in Doppler flow velocities. The condition of our patients most likely was related to a transient inflammation (or thickening) of the pericardium due to viral, bacterial, or immunologically mediated pericarditis. Resolution of the thickened pericardium was documented by magnetic resonance imaging in one patient. Awareness of the possible transient nature of constrictive pericarditis in a subgroup of patients with constriction has important clinical implications when pericardiectomy is considered. The resolution of constrictive pericarditis can be documented by serial Doppler echocardiographic examination. PMID- 8246617 TI - Clinical performance of an extraction calcitonin radioimmunoassay. AB - An extraction radioimmunoassay of calcitonin (CT) was used to replace the direct radioimmunoassay, in which false-negative results (normal basal CT values but above normal ["positive"] results after pentagastrin stimulation) occurred in 26 to 30% of cases. During the past 6 years, 170 pentagastrin stimulation tests were performed in 135 patients at our institution. We reviewed the results to determine whether extraction radioimmunoassay yielded improvement. Of the 135 patients, 6 (4.4%) had indeterminate results of pentagastrin stimulation tests that consisted of normal basal levels but increased values after pentagastrin stimulation, a considerable improvement over the results with direct assay. Six other patients had indeterminate results of pentagastrin testing--increased basal CT levels but normal values after pentagastrin stimulation. Indeterminate results of pentagastrin tests occurred primarily in postoperative or preoperative patients with a family history of medullary thyroid carcinoma (MTC). The reasons for the indeterminate results were unclear, as was their importance as a prelude to disease, but two preoperative patients with indeterminate results eventually had positive results and were found to have MTC at operation. We conclude that extraction radioimmunoassay is more reliable than direct radioimmunoassay for the diagnosis of MTC. Determination of a basal CT value and of the CT value after pentagastrin stimulation is sufficient for diagnosing MTC. PMID- 8246618 TI - Posture Training Support: preliminary report on a series of patients with diminished symptomatic complications of osteoporosis. AB - Back supports are often used to minimize or prevent complications of osteoporosis. Nevertheless, the few related studies to date reveal that the currently available thoracolumbar and lumbosacral supports have substantial limitations, including (1) poor compliance because of discomfort or restricted motion, (2) expense, (3) unacceptable cosmetic and aesthetic appearance, and (4) medical contraindications to the use of rigid supports. We report the initial results of a clinical trial of the Posture Training Support (a thoracolumbar support) in 29 women and 1 man with osteoporosis or osteopenia of the spine (ages 37 to 87 years), who were referred because of back pain or kyphosis. We hypothesize that this inexpensive, unobtrusive device promotes improvement in posture and reduces back pain either by acting as a proprioceptive reinforcer or by producing a force posteriorly below the inferior angles of the scapulae and thus decreasing the anterior compressive forces that are commonly exerted on the spine. Among the 23 patients who reported substantial back pain before use of the support, relief of the pain was "significant" in 17 and minimal in 6. Nineteen patients noted improvement in their posture. No patient reported worsening of back pain or posture, nor did any patient discontinue use of the device for cosmetic reasons, discomfort, or other complaints. Four patients previously could not tolerate other back supports, and 14 had previously used other supports without substantial improvement. These preliminary results suggests that the Posture Training Support may be of considerable symptomatic and prophylactic value to patients with osteoporosis who cannot tolerate conventional back supports. PMID- 8246619 TI - Charles Barkla--Nobel Laureate. PMID- 8246620 TI - Development of pulmonary hypertension after placement of a ventriculoatrial shunt. AB - The ventriculoatrial shunt (VAS) was developed to control hydrocephalic syndromes effectively. Several complications, however, have been described after the procedure. One of the most serious consequences is the development of severe pulmonary hypertension attributed to multiple and recurrent pulmonary embolization caused by the catheter of the VAS; however, the frequency is exceedingly low. Herein we describe the experience with three patients in whom severe pulmonary hypertension developed after a VAS procedure. In two patients, refractory heart failure developed, an outcome that caused death within a brief period. The third patient underwent atrial thrombectomy and then pulmonary thromboendarterectomy; recovery was complete. Scientific evidence shows that initial embolization predisposes pulmonary vessels to develop further in situ thrombosis; thus, the vascular lung disease progresses despite removal of the embolic source. A review of the literature revealed that in patients with a VAS, pulmonary embolism and pulmonary hypertension were clinically diagnosed in only 0.4% and 0.3% of the cases, respectively, whereas postmortem diagnoses of pulmonary embolism and pulmonary hypertension were established in 59.7% and 6.3%, respectively. These discrepancies point out the difficulty of establishing the diagnosis of these serious pulmonary vascular complications while the patient is alive. PMID- 8246621 TI - Hydroxyapatite orbital implant after enucleation--experience with 200 cases. AB - The recently introduced hydroxyapatite orbital implant was designed to provide improved motility of the prosthesis in patients who have undergone enucleation. To date, little has been written about the use of the hydroxyapatite orbital implant in eyes that harbored malignant neoplasms. We have performed hydroxyapatite orbital implantation in approximately 200 patients, including children as young as 1 month of age, who underwent enucleation primarily for malignant intraocular tumors. During a median follow-up of 24 months, no cases of orbital hemorrhage, orbital infection, implant extrusion, or implant migration have occurred. Three patients have had conjunctival erosion, which was successfully repaired. Motility of the socket and fornices has been excellent, and all patients have cosmetically satisfactory motility of the prosthesis. Use of the hydroxyapatite implant has had no adverse effect on the clinical course of the intraocular neoplasms, and it has not hindered postoperative evaluation for recurrent orbital tumor. Overall, the hydroxyapatite orbital implant seems to be well tolerated, provides improved motility of the prosthesis, and is associated with few complications. PMID- 8246622 TI - Current management of posterior uveal melanoma. AB - The management of malignant melanoma of the ciliary body and choroid (posterior uvea) is controversial. Authorities have disagreed about whether enucleation or conservative treatment offers the best prognosis. Although retrospective studies have suggested that the method of treatment makes no difference in the systemic prognosis, new studies in which the various therapeutic modalities are being compared are currently under way. The Collaborative Ocular Melanoma Study is attempting to address some of these issues in a randomized clinical trial. In this report, the currently available methods for managing posterior uveal melanoma are reviewed. Small asymptomatic choroidal melanomas can probably be observed periodically until evidence of growth is documented. Some small choroidal melanomas can be treated with laser photocoagulation. Alternatively, radiotherapy (either episcleral application of a radioactive plaque or charged particle irradiation) can be used. Although the two methods of radiotherapy seem equal relative to the development of systemic metastatic lesions, plaque radiotherapy is associated with fewer and less severe local complications. Selected melanomas of the ciliary body and peripheral choroid can be treated by local resection (partial lamellar sclerouvectomy). Local resection has theoretical advantages, but the surgical procedure is associated with potentially greater immediate complications. Enucleation is generally indicated for advanced melanomas that occupy most of the intraocular structures or have caused severe glaucoma. In addition, it is usually recommended for tumors that have invaded the optic nerve. The value of preenucleation radiotherapy in improving patient survival is unproved, although this technique seems reasonable in selected advanced tumors in which enucleation seems inevitable. Orbital exenteration is justified for advanced uveal melanomas with massive extraocular extension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246623 TI - Mesenteric volvulus. PMID- 8246624 TI - Cyclophosphamide-induced hepatotoxicity in a patient with Wegener's granulomatosis. AB - Cyclophosphamide is widely used in the treatment of many vasculitides. Hepatotoxicity associated with such therapy is uncommon but possible. Herein we describe a well-substantiated case of cyclophosphamide-induced hepatotoxicity in a patient with Wegener's granulomatosis. Physicians should be aware of this potentially serious reaction when cyclophosphamide therapy is initiated. Baseline liver function tests and periodic assessment are recommended during treatment with cyclophosphamide. PMID- 8246625 TI - Prolonged remission of metastatic follicular thyroid carcinoma. AB - Follicular thyroid carcinoma is associated with an extremely poor prognosis when metastatic lesions occur. Although radioiodine therapy for metastatic disease can extend patient survival for several years, its effect is usually only palliative. Herein we describe a 47-year-old man with metastatic follicular thyroid carcinoma who was in clinical remission for 30 years after surgical and radioiodine treatment of the original disease. To our knowledge, this is the longest reported remission of this disease. Possible reasons for the prolonged survival were "young" age at diagnosis, diploid DNA content of the tumor, skeletal lesions responsive to 131I therapy, and limited pulmonary disease. PMID- 8246626 TI - Testing for arsenic. PMID- 8246627 TI - Use and abuse of medical service marks. AB - Medical service marks, like other service marks and trademarks, are subject to public misuse and infringement. Such misuses are sometimes innocent and sometimes fraudulently motivated. For example, throughout the history of the Mayo Clinic, the Mayo name has been publicly appropriated by unauthorized users attempting to claim an endorsement or affiliation with the clinic. On at least two occasions, the Mayo Clinic has sued misusers. Mayo prevailed in a 1962 appeal in the Minnesota Supreme Court against a business incorporating into its name the word "Mayo" and selling medicinal products in the Rochester, Minnesota, area. The supreme court banned such deceptive use, finding that persons would associate the name Mayo on medically related products with the Mayo Clinic. Mayo did not prevail, however, in a 1972 federal appeal against a food company attempting to register a trademark of "mayo" and "7" for mayonnaise. The court found that purchasers would not associate the "mayo" on a food product with the Mayo Clinic. From 1989 to 1991, a fraudulent "Mayo Diet Pill" circulated in Europe, where advertisements suggested that it originated at the Mayo Clinic. Its sale was stopped only after the Mayo reputation incurred an undetermined amount of damage in Europe. Public misuse of medical service marks is likely to increase as the health-care marketplace becomes more competitive. PMID- 8246628 TI - Congestive heart failure: insights from epidemiology, implications for treatment. PMID- 8246629 TI - Rhinoscleroma in perspective. PMID- 8246630 TI - Electronic manipulation to enhance medical photographs. PMID- 8246631 TI - Former editors just fade away. PMID- 8246632 TI - Familial co-segregation of the elastin phenotype in skin fibroblasts from Hutchinson-Gilford progeria. AB - Elastin and type IV collagen production are markedly elevated in fibroblasts derived from the skin of patients with Hutchinson-Gilford progeria (HGP). Fibroblasts from three affected children and their parents were compared to normal human skin fibroblasts with respect to elastin production as a function of different concentrations of calf serum and the cytokines, transforming growth factor-beta and basic fibroblast growth factor (TGF-beta 1, bFGF). In cultured fibroblasts from the parents of probands that were very high elastin producers (> 10(5) molecular equivalents/cell per h), at least one parent (mother) presented the same phenotype. Overproduction of elastin in culture could have been due to increased sensitivity of HGP strains to stimuli present in serum; however, relative stimulation of elastin production by calf serum in cell strains from HGP elastin over-producers was less than half the control strain. In most of the cultures examined, the responsiveness of elastin production to TGF-beta 1 was almost absent when compared to the response of normal fibroblasts. HGP strains with high elastin production modified conditioned medium to enhance elastin production in normal cells. These results suggest the presence, in HGP skin fibroblasts, of inheritance of high elastin production that is associated with accelerated aging. PMID- 8246633 TI - Maximum life span in vertebrates: relationship with liver antioxidant enzymes, glutathione system, ascorbate, urate, sensitivity to peroxidation, true malondialdehyde, in vivo H2O2, and basal and maximum aerobic capacity. AB - In order to help clarify whether free radicals are implicated or not in the evolution of maximum life span (MLSP) of animals, a comprehensive study was performed in the liver of various vertebrate species. Strongly significant negative correlations against MLSP were found for hepatic catalase, Se-dependent and -independent glutathione peroxidases, and GSH, whereas superoxide dismutase, glutathione reductase, ascorbate, uric acid, GSSG/GSH, in vitro peroxidation (TBA RS), and in vivo steady-state H2O2 concentration in the liver did not correlate with MLSP. Superoxide dismutase, catalase, glutathione peroxidase, and GSH results were in agreement with those independently reported by other authors, whereas the rest of our data are reported for the first time. Potential limitations arising from the use of animals of different vertebrate Classes were counterbalanced by the possibility to study animals with very different MLSPs and life energy potentials. Furthermore, the results agreed with previous data obtained using only mammals. Since liver GSSG/GSH, peroxidation, and specially H2O2 concentration were similar in species with widely different MLSPs, it is suggested that the decrease in enzymatic H2O2 detoxifying capacity of longevous species represents an evolutionary co-adaptation with a smaller in vivo rate of free radical generation. We propose the possibility that maximum longevity was increased during vertebrate evolution by lowering the rate of free radical recycling in the tissues. PMID- 8246634 TI - Age-dependent modifications of mitochondrial trans-membrane potential and mass in rat splenic lymphocytes during proliferation. AB - The specific fluorescent probes, Rhodamine 123 (Rh-123) and Nonyl-Acridine Orange (NAO) were, respectively, used to monitor the changes in membrane potential and mass of lymphocyte mitochondria during aging and proliferation. An age-dependent increase of the uptake of both fluorochromes was observed in resting cells; however, NAO fluorescence increased to a greater extent when compared with the Rh 123 probe. This resulted in a lower respiratory activity per unit of mitochondrial mass in old cells than in the young ones. Following mitogenic stimulation, most of the lymphocytes from young rats showed an increase in their membrane potential and mass. On the contrary about 50% of cells from old rats had depolarized mitochondria after 72 h from the stimulation. Present data support that mitochondria of lymphocytes from old rats are extremely sensitive to the stressing conditions resulting from mitogenic stimulation. PMID- 8246635 TI - Evidence for accelerated skin wrinkling among developmentally delayed individuals with Down's syndrome. AB - Down's Syndrome (DS) is a genetic disorder involving mental retardation which is reported to be a condition of accelerated aging. However, few studies exist which examine age-dependent changes in DS and these studies fail to include a control group. In the present study a non-invasive, painless, and easily obtainable measure reflecting age-dependent topographical changes of the skin (skin wrinkling) was obtained in 15 DS subjects, 14 developmentally disabled (DD) (mentally retarded) controls (DD controls) and 16 healthy controls of similar ages ranging from 22 to 51 years. The healthy control group failed to show correlations of skin measures with age, as predicted for the age-range of these subjects, based on results of previous studies. However, the DS group showed significant correlations of all skin measures from relatively non-sun-exposed and sun-exposed skin sites, with age. The DD control also showed significant correlations with age of several but not all skin sites and the correlation coefficients tended to be less than that of the DS; at least, when contrasted with the healthy controls the DS showed greater values of several skin measures suggestive of accelerated skin wrinkling. These results provide evidence for accelerated aging of the skin, and possibly greater effects of sun-exposure on skin wrinkling, in DS and possibly (DD) individuals in contrast to healthy individuals without a genetic disorder and without mental retardation. PMID- 8246636 TI - HnRNP CBP35.CBP67 interaction during stress response and ageing. AB - Previous studies have demonstrated the existence of nuclear carbohydrate binding proteins in a variety of mammalian cells with molecular masses of 35,000, 67,000, and 70,000 (CBP35, CBP67, and CBP70), which are associated with nuclear ribonucleoprotein (RNP) complexes. CBP35 consists of two domains, an amino terminal portion that is homologous to certain regions of proteins of the heterogeneous nuclear RNP complex, and a carboxyl-terminal portion homologous to beta-galactoside-specific lectins. CBP35 it has been proposed, like the glucose specific lectin, CBP67, to guide RNP complexes through the nuclear pore. Here we show that the exposure of mature rats to stress induces an increase in nuclear CBP35 bound to CBP67 and retained on immobilized glucose. Nuclear extracts from the livers of old rats displayed no detectable stress response. This CBP35.CBP67 association detected in rat liver is considered with respect to the CBP35.CBP70 association recently observed in HL60 cell nuclear extracts. PMID- 8246637 TI - Drug utilization review in ambulatory settings: state of the science and directions for outcomes research. AB - There are escalating national pressures to analyze pharmaceutical outcomes and to develop drug-related clinical guidelines. These interests coincide with passage of the Medicaid Rebate Law (OBRA, 1990), which mandates the implementation of prospective and retrospective drug utilization review (DUR) programs by Medicaid in 1993. This report investigates DUR programs that target outpatient drug therapies. The authors present a conceptual framework that identifies the factors influencing drug prescribing and the range of potential patient outcomes. Current types of DUR interventions and their applications are described, in addition to problems that hinder implementation or evaluation of DUR programs. DUR evaluation studies are reviewed, and a critique identifies the limitations of available DUR research. The authors recommend an expanded DUR policy research agenda, strongly suggesting that priority be given to studies in the following areas: DUR criteria development and validation; prevalence of prescribing problems and their association with patient outcomes; efficacy, toxicity and costs of therapeutic alternatives; and DUR program evaluation. The overall conclusion is that the state of the science pertaining to DUR is not well developed. The potential of DUR may not be realized due to the lack of resources needed to design, implement, and evaluate effective programs. Instead, DUR efforts may be limited to cost containment issues without due consideration of quality-of-care outcomes. The authors call for rigorous evaluation efforts to inform DUR design and implementation, thereby assuring more rational prescribing and enhancing patient outcomes. PMID- 8246638 TI - Physician satisfaction with primary care office visits. Collaborative Study Group of the American Academy on Physician and Patient. AB - To study encounter-specific physician satisfaction we collected exit questionnaires from patients and physicians following 550 primary care office visits. The physicians' questionnaire included 20 items pertaining to satisfaction with the visit, one of which was an assessment of global satisfaction. Using a boot-strap technique, we factor analyzed the satisfaction questions in 10 repeated samples. Four distinct dimensions of physician satisfaction emerged: satisfaction with the patient-physician relationship, with the data collection process, with the appropriateness of the use of time, and with the absence of excessive demands on the part of the patient. Each scale was found to be reliable; global satisfaction was most closely related to the relationship factor. Satisfaction with use of time and the adequacy of data collection tended to be stable for individual physicians across a range of patients whereas global satisfaction and satisfaction with the relationship and the demanding nature of the patient and were more variable, hence most unique to each encounter. This study of physician satisfaction represents an effort to incorporate knowledge about physicians' subjective experiences into a systematic understanding of the dynamics of the medical interview. PMID- 8246639 TI - Who enrolled in a state program for the uninsured: was there adverse selection? AB - Managed care plans may hesitate to participate in programs for uninsured persons because they fear adverse selection, whereby only the sickest people or highest users would choose to join the program. We studied this issue in Washington State's Basic Health Plan, a demonstration program that provides subsidized health insurance for families earning less than 200% of the poverty level. We interviewed people in three counties who enrolled in the program, and compared them to people in the same counties who were eligible but did not enroll. There were substantial differences between enrollees and eligibles in education, age, income, employment, race, and insurance status. In spite of these demographic and access differences, health status was remarkably similar for enrollees and eligibles, with the few significant differences favoring the enrollees. In addition, previous and subsequent use of health services was similar or lower for enrollees. The results for health status and utilization were similar across the three counties, even though the counties and the providers were quite different. We conclude that there is no evidence of adverse selection. This is welcome news for the health plans, but suggests that the BHP may not have reached those most in need of insurance. PMID- 8246640 TI - Would eliminating differences in physician practice style reduce geographic variations in cataract surgery rates? AB - This study uses Medicare physician-claims data to examine patient and physician contributions to variations in cataract surgery rates across U.S. metropolitan areas. Utilization is modelled as having two phases: the decision to seek an ophthalmologist's care, which is made by patients, and the decision to perform surgery on patients who seek care, which is partially controlled by ophthalmologists. Under this model, the effect of physician practice style on cataract surgery rates occurs through the influence of practice style on the second phase of utilization. Variation in patient care-seeking behavior contributed to the variation in the rate of cataract surgery. Moreover, multivariate regression analyses found that cataract surgery rates were influenced by economic and sociodemographic variables in predictable ways. Using the regression results, a "purged" cataract surgery rate that was free of any possible influence of physician practice style was calculated. Variation in the purged surgery rate was only slightly lower than variation in the observed surgery rate, suggesting that eliminating practice style as a factor in physician decision making (e.g., through practice guidelines) would reduce variations in cataract surgery rates by only a small amount. PMID- 8246641 TI - Patient-mix differences among ambulatory providers and their effects on utilization and payments for Maryland Medicaid users. AB - Characteristics of the usual source of care (e.g. specialty, organizational type) are known to be related to utilization and cost levels. This study assesses the degree to which variations in utilization and cost are attributable to differences in patient mix (i.e. demographic and diagnostic characteristics). Comparisons are made with Medicaid payments with and without patient-mix adjustment among users of hospital outpatient departments, emergency rooms, Federally Qualified Health Centers and office-based physicians. The study builds upon previous work by including a well-developed set of ambulatory case-mix controls, a variety of provider types, a large number of providers, and a relatively comprehensive resource utilization component. Findings confirm significant differences in patient demographic and diagnostic characteristics among users of different types of providers. Controlling for these patient-mix characteristics explains 44% of the variation in ambulatory use, 21% in hospital admissions, and 13% in total Medicaid payments. The considerable remaining variation suggests differences in provider efficiency. For example, even after patient mix adjustment, 18% of those who rely on outpatient departments are hospitalized annually compared to 10% for users of office-based physicians. Overall findings indicate that patient-mix characteristics, as well as differences in provider efficiency, should be considered when developing and evaluating managed care and preferred provider initiatives for ambulatory care. PMID- 8246642 TI - The importance of patient preferences in the measurement of health care satisfaction. AB - The idea that patients will be more satisfied with health care services that are delivered to meet their preferences is central to the concept of health care marketing. Health care providers increasingly use market segmentation and target marketing to optimize the fit between their services and the consumers who receive them. This study evaluates one model for incorporation of patient preferences into the measurement of satisfaction. Using multiple regression analysis, evaluations of three dimensions of health care satisfaction, interpersonal care, technical quality, access to care accounted for 63% of the variance in overall satisfaction. Inclusion of preferences, defined as importance ranks of each dimension, did not improve ability to predict satisfaction. Four preference segments were identified: interpersonal care seekers, access/quality seekers, access seekers and quality seekers. These four subgroups differed significantly on a number of sociodemographic, health status and health service use characteristics but no significant differences were found in satisfaction between preference segments. Patient satisfaction can best be measured as quality evaluations of dimensions without regard to preferences. In considering the merits of market segmentation and target marketing, alternative satisfaction models that link preferences to health care satisfaction or the possibility that preference targeting does not lead to greater satisfaction should be evaluated. PMID- 8246643 TI - Typhus and its control in Russia, 1870-1940. PMID- 8246644 TI - Palmitate or permanganate: the venereal prophylaxis debate in Britain, 1916-1926. PMID- 8246645 TI - Epidemiology of puerperal fever: the contributions of Alexander Gordon. PMID- 8246646 TI - British Army surgeons commissioned 1840-1909 with west Indian/west African service: a prosopographical evaluation. PMID- 8246647 TI - Independent discovery in biology: investigating styles of scientific research. PMID- 8246648 TI - Illustrations from the Wellcome Institute Library. Nicholas Gainsford: his book. PMID- 8246649 TI - Surgery for vertigo in the nonserviceable hearing ear: transmastoid labyrinthectomy or translabyrinthine vestibular nerve section. AB - Two of the surgical options that exist for the treatment of disabling vertigo arising from an ear with nonserviceable hearing are a transmastoid labyrinthectomy (TL) and a translabyrinthine vestibular nerve section (TLVNS). The major difference between the two operations is a section of the vestibular nerves with the TLVNS which removes all preganglionic vestibular tissue from the diseased inner ear. It has been inferred that a TLVNS should be the procedure of choice if hearing is not to be spared, because a TL results in an incomplete removal of preganglionic vestibular tissue, and that this remaining tissue might have continued or recurrent physiologic function resulting in further vertigo. The clinical outcome of 58 patients who had either TL or TLVNS for disabling vertigo arising from a nonserviceable hearing ear was investigated with respect to the control of vertigo and the development of postoperative balance dysfunction. The control of vertigo in the TLVNS and TL groups was 100% and 95.3%, respectively. This difference was not statistically significant. There was a tendency for postoperative dysequilibrium to be more frequent in the TLVNS group, but this finding did not reach statistical significance. A TL appears to offer the same benefit as TLVNS in the control of intractable episodic vertigo without the additional risks of TLVNS. PMID- 8246650 TI - Idiopathic facial nerve paralysis: a randomized double blind controlled study of placebo versus prednisone. AB - Idiopathic facial nerve paralysis (IFNP) is a common malady. Because its etiology is unclear, there are a variety of treatment options. Studies to date have not clearly established the benefits of treatment with oral steroids (prednisone). The authors performed a randomized double-blind controlled study comparing the use of placebo versus prednisone which shows that prednisone-treated patients benefit from early treatment. Seventy-six patients met inclusion criteria and completed follow-up until recovery; 35 patients received prednisone and 41 received placebo. Their mean age was 36.8 years. Facial nerve function was assessed using the House-Brackmann facial nerve grading scale, as well as a variety of other measures. Patients were evaluated pretreatment, regularly post treatment until judged recovered (return of facial function to a grade III or better), and at 6 months after recovery. Difference in mean time to resolution for the prednisone (51.4 days) and placebo (69.3 days) groups was not statistically significant. There was a significant difference in grade at recovery, with the placebo group having a higher proportion of grade III results (P < .03). Eight of 10 patients with electroneurography (ENOG) evidence of denervation were in the placebo group and accounted for 6 of the 7 grade III results. However, the difference in proportion of patients with evidence of denervation for the prednisone (5.7%) and placebo (19.5%) groups did not achieve statistical significance. This study shows that patients treated with prednisone have less denervation than placebo-treated patients. They also have a significant improvement in facial grade at recovery compared to placebo-treated patients. Therefore, the authors recommend that all patients at risk for developing denervation receive prednisone treatment. PMID- 8246651 TI - Otoacoustic emissions in full-term newborns at risk for hearing loss. AB - Distortion product otoacoustic emissions (DPOEs) and click-evoked otoacoustic emissions (CEOEs) characteristics of the normal newborn population have been previously reported in the literature. There is little information about DPOE evaluations in the newborn population at risk for hearing loss. The authors now report the DPOE and/or CEOE data from six full-term subjects at risk for hearing loss or with highly suspected hearing loss. These subjects were less than 1 year of age and at risk for hearing loss secondary to a history of hereditary hearing loss, meningitis, hyperbilirubinemia, and ototoxic drug exposure. Audiometric evaluation included auditory brainstem responses (ABR), behavioral observation audiometry, and tympanometry. The CEOEs and DPOEs were found to be decreased or absent in the subjects with suspected hearing loss secondary to cochlear pathology; they were found to be normal in a subject with a suspected central hearing loss. This study's data suggest that otoacoustic emissions when combined with ABR can provide a frequency-specific evaluation of cochlear function and help determine the anatomic site of a pathologic lesion. PMID- 8246652 TI - Reclassification of aggressive adenomatous mastoid neoplasms as endolymphatic sac tumors. AB - The emerging concept that aggressive adenomatous tumors of the temporal bone arise from the endolymphatic sac and constitute a distinct clinicopathologic entity merits wider recognition. These tumors share a common clinical pattern and exhibit consistent imaging and histopathologic features. Endolymphatic sac tumors (ELSTs) have been mistaken for other neoplasms such as paragangliomas, adenomatous tumors of mixed histology, ceruminomas, and choroid plexus papillomas. A review of the literature shows similarities among case studies of these aggressive adenomatous lesions. An analysis of the data supports the endolymphatic sac as an origin for these tumors. This report also presents an additional case of a less differentiated variant of this rare but important clinicopathologic entity. PMID- 8246653 TI - The effect of tonsillectomy and adenoidectomy on neutrophil chemotaxis. AB - Although tonsillectomy and adenoidectomy are common surgical procedures, the effects of these operations on the immune system have not been thoroughly determined. Our data on neutrophil chemotaxic functions in a group of 17 patients with chronic tonsillitis and adenoid hypertrophy show that chronic tonsillitis and adenoid hypertrophy impair neutrophil chemotaxic functions and that there is a subsequent normalization of these values following tonsillectomy and adenoidectomy. PMID- 8246654 TI - Composite graft tympanoplasty in the treatment of ears with advanced middle ear pathology. AB - In ears with advanced pathology, the functional and anatomical results of surgery are compromised by such factors as total perforation, tympanosclerosis, atelectasis, suppuration, or previous surgery. Perichondrium cartilage composite grafts (PCCGs) were used for membrane grafting in 550 such cases over a period of 5 years. In all, 197 tympanoplastic procedures of type I (25% with previous surgery) and 353 of type III (47% with previous surgery) were carried out. The graft was obtained from the tragus and/or the concha. Two slightly different surgical techniques of grafting were used. Closure of the eardrum perforation was successful in 92% of the procedures. An air-bone gap of less than or equal to 30 dB was obtained in 92.4% of tympanoplasty type I procedures and in 79% of tympanoplasty type III procedures. The conclusion drawn from the results is that PCCG is a reliable graft in cases of advanced destruction of the middle ear. PMID- 8246655 TI - Clinical signs of orbital ischemia in rhino-orbitocerebral mucormycosis. AB - Mucormycosis is the most acutely fatal fungus infection of man (Ferry and Abedi). The most common clinical type of infection is rhino-orbitocerebral mucormycosis. Prompt recognition of the clinical picture is essential if the appropriate urgent management is to be instituted without delay. The presence of black eschar in the region of the nasal passages, palate, midface, and orbit is the best-recognized clinical sign alerting the clinician to the diagnosis. Black eschar is, however, a feature in only a minority of these patients at the time of presentation. This paper discusses other clinical signs, particularly orbital ischemia, which should suggest the diagnosis. The clinical presentation of orbital ischemia in mucormycosis includes proptosis, total external and internal ophthalmoplegia, and early blindness. A lax, nontender periorbital puffiness, which does not feel warm to the examiner's touch, is typical. Proptosis and chemosis, if present, are mild. These signs are compared with those of pyogenic orbital cellulitis, with which the condition might most easily be confused. PMID- 8246656 TI - Rhabdomyosarcoma of the head and neck in adults: a study of 26 patients. AB - The clinical records of 26, predominantly male, adults with rhabdomyosarcomas in the head and neck were analyzed. Patients' ages ranged from 18 to 74 years (mean: 24.5 years). According to the retrospective clinical group classification, 18 (69%) of 26 were advanced tumors at initial presentation belonging to group III or IV. The ethmoids were the most common primary site of origin in 12 (46%) of 26 patients. Nodal and systemic metastases were noted in 12 (46%) and 6 (23%) patients, respectively. Bone metastases were noted in 4 patients. Heterogeneous treatment protocols were used with a variety of chemotherapy combinations in most cases, with surgery and radiotherapy. Overall results were poor, with a survival rate of 7.6% at 5 years. Neither histopathology nor response to chemotherapy was found to influence survival. All long-term survivors belonged to the early-stage groups (clinical groups I and II) for which complete surgical excision was possible. In spite of a poor prognosis after relapse, the use of aggressive chemotherapy appeared to prolong life in some patients. PMID- 8246657 TI - Airway resistance and work of breathing in tracheostomy tubes. AB - Recommendations for sizing of tracheostomy tubes are generally based on anatomic considerations with the largest fitting tube most commonly placed. Once in the tracheostomy site, the tube assumes the new role of the upper airway. Consideration of the airway resistance of each tracheostomy tube and change in work of breathing are important in maintaining the respiratory system homeostasis. The airflow dynamics of neonatal, pediatric, and adult tracheostomy tubes were studied. Flow rates were plotted against change in pressure for inspiratory and expiratory flows and resistances for each tube were calculated. The expiratory resistances were larger for the neonatal tubes and pediatric tubes 0 and 00, while inspiratory resistances were the limiting factor in the adult tubes and the larger pediatric tubes. Comparison of calculated resistances of the tracheostomy tubes was made with known physiologic airway resistances. Adult tubes 8 and 10 most closely simulated the upper airway resistance of adults and neonatal tube 0 appeared most appropriate for the newborn. Work of breathing was determined for each tracheostomy tube. Increasing tube diameter as well as decreasing tidal volume and respiratory rate decreased the amount of work required to maintain a given flow. PMID- 8246658 TI - Supracricoid hemilaryngopharyngectomy in selected pyriform sinus carcinoma staged as T2. AB - From 1964 to 1985 supracricoid hemilaryngopharyngectomy (SCHLP) was performed at the authors' institution for 34 selected pyriform sinus carcinomas staged as T2. Tumors involved the anterior part of the pyriform sinus, the lateral wall, the medial wall, and the whole aryepiglottic fold in all cases. Tumors with invasion of the apex of the pyriform sinus, of the retrocricoid region, of the posterior pharyngeal wall, or with fixation of the true vocal cord were excluded from the study. Such a technique was aimed at preserving physiologic phonation, respiration, and swallowing while achieving the same local control rate as pharyngolaryngectomy. Patients were monitored for at least 6 years or until death. No patients were lost to follow-up. The 5-year cause-specific survival rate was 55.8%. The main cause of death was second primary tumor. The 5-year actuarial local recurrence rate was 3.4%. The authors' experience with the SCHLP technique challenges the traditional teaching of pharyngolaryngectomy and establishes this technique as a safe method of voice preservation in selected cases of pyriform sinus carcinomas. PMID- 8246659 TI - The use of cartilage pate in rebuilding mastoidectomy cavities: an alternative method. PMID- 8246660 TI - Otosclerosis regrowth. PMID- 8246661 TI - Juvenile aggressive cemento-ossifying fibroma: case report and review of the literature. PMID- 8246662 TI - Improvements in the assessment of pulmonary function in laryngectomized patients. PMID- 8246663 TI - Modified epiglottoplasty for prevention of aspiration. PMID- 8246664 TI - Stabilization of craniofacial prostheses using osseointegrated titanium implants. PMID- 8246665 TI - Directory of Otolaryngological Societies. PMID- 8246666 TI - L-arginine inhibits neointimal formation following balloon injury. AB - Nitric oxide (NO)-generating vasodilators inhibit the mitogenesis and proliferation of cultured vascular smooth muscle cells. We investigated the role of NO in the vascular response to arterial injury by administering L-arginine (precursor of NO), D-arginine or N-nitro L-arginine methylester (NAME; an inhibitor of NO synthesis) to a rat model of balloon catheter-induced left carotid artery injury. Two weeks after the balloon injury, animals that received both oral (1.25 g/l water) and local (10mg in gel) administration of L-arginine showed suppression of neointimal proliferation with no change in systolic blood pressure. Medial proliferation was potentiated in NAME-treated animals with a higher blood pressure. Tissue cGMP content (representative of NO generation) of the injured arteries was similar to that of normal arteries with intact endothelium. These findings suggest that a higher local concentration of NO produced from L-arginine can inhibit the migration and proliferation of smooth muscle cells in the injured vascular wall. PMID- 8246667 TI - Stress alters adenylyl cyclase activity in the pituitary and frontal cortex of the rat. AB - We hypothesized that sustained biosynthesis of proopiomelanocortropin (POMC) from the anterior pituitary during chronic stress might result in enhanced membrane adenylyl cyclase (AC) activity, facilitating amplification of the CRH signal despite falling numbers of CRH receptors. Therefore, we investigated the effects of stress on AC activity in anterior pituitaries from Sprague-Dawley rats exposed to stress. Following 12 h of intermittent, cold, swim stress, stressed rats had plasma corticosterone levels that were 10 fold higher than in nonstressed animals and showed a 40% reduction in the specific binding of 125I-CRH to anterior pituitary membranes. Moreover, stressed rats showed a 3 fold increase in anterior pituitary POMC mRNA levels. To test the hypothesis that factors released during stress enhanced the AC signal transduction system, thereby leading to increased POMC gene expression, we measured anterior pituitary cAMP and assayed AC activity from membranes prepared from anterior pituitary of control and stressed rats. Levels of cAMP were 2 fold higher in pituitaries from stressed rats compared to controls. The significant increase in cAMP was accompanied by a significant increase of AC activity. To test what component(s) of the AC complex are altered by stress, type I and II AC mRNA as well as Gs alpha, Gi(1-3) alpha and G beta protein levels were determined. Type II AC mRNA was significantly increased 1.7 fold in stressed rats compared with controls, whereas no consistent alteration in G-protein levels were detected. Enhanced AC activity following cold swim stress was not limited to the pituitary, to one line of rat, nor one type of stress. In Fisher rats, both cold swim and restraint stress enhanced AC activity in the pituitary and in the frontal cortex. In summary, stress enhances AC activity in the anterior pituitary. The increase in AC activity is associated with increased steady state levels of type II AC mRNA. Factor(s) released during stress may enhance AC signal transduction and allow for persistent elevation in POMC gene expression despite the inhibitory influences of glucocorticoids. PMID- 8246668 TI - Gonadotropin-induced ovarian carbonyl reductase in mice and hamsters: comparison with carbonyl reductase in rats. AB - We investigated the effects of pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) on ovarian carbonyl reductase activities towards 13,14-dihydro-15-ketoprostaglandin F2 alpha (15KD-PGF2 alpha), p nitroacetophenone (PNAP) and p-nitrobenzaldehyde (PNBA) in mice and hamsters, and compared with their effects on those we observed previously in rats. The treatment with PMSG and hCG caused a significant increase in ovarian weights and superovulation in both mice and hamsters. Hamster ovary possessed appreciable carbonyl reductase activities towards all three substrates, whereas the activities were lower than those in rat ovary. The reductase activities were not increased by the treatment with gonadotropins, differing from rat ovarian carbonyl reductase. In untreated mice, carbonyl reductase activity towards 15KD PGF2 alpha was not detected, whereas the activities towards PNAP and PNBA were detected, which activities were lower than those in rats and hamsters. The PNAP and PNBA reductase activities in mouse ovary were significantly increased up to 7.1- and 1.7-fold, respectively, by the treatment with gonadotropins. These results show that there are species differences in ovarian carbonyl reductase and response of the enzyme to gonadotropins. PMID- 8246669 TI - Corticotropin-releasing hormone modulates cytokines release in cultured human peripheral blood mononuclear cells. AB - Immune and neuroendocrine systems interact at various levels. In particular, either cytokines activate the hypothalamus-pituitary-adrenal axis (HPA) or corticotropin-releasing hormone (CRH) induces the release of beta-endorphin from peripheral human mononuclear cells. The aim of the present study was to investigate whether CRH may affect cytokine production and activity in human peripheral blood mononuclear cells (PBMC). Primary cultures of human PBMC and monocytes were used. They were incubated in presence of different doses of synthetic human CRH. Media were collected and interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) levels were measured by ELISA, while interferon-gamma (IFN gamma) levels were measured by bioassay. In addition, phytohemoagglutinin-induced lymphocyte proliferation was evaluated by testing [3H]thymidine incorporation in the presence of various doses of CRH. CRH significantly increased IL-6 release from PBMC (p < 0.01). The addition of CRH to PBMC significantly decreased IFN gamma levels, in a dose dependent manner (p < 0.01). No significant effect of CRH was observed on lymphocyte proliferation or IL-1 beta production. The present results suggest a role for CRH as a paracrine mediator for human immune cells, increasing the evidence of a clear correlation between immune and neuroendocrine system. PMID- 8246670 TI - A novel muscarinic receptor ligand which penetrates the blood brain barrier and displays in vivo selectivity for the m2 subtype. AB - Alzheimer's disease (AD) involves selective loss of muscarinic m2, but not m1, subtype neuroreceptors in the posterior parietal cortex of the human brain. Emission tomographic study of the loss of m2 receptors in AD is limited by the fact that there is currently no available m2-selective radioligand which can penetrate the blood-brain barrier. In our efforts to prepare such a radioligand, we have used competition studies against currently existing muscarinic receptor radioligands to infer the in vitro and in vivo properties of a novel muscarinic receptor ligand, 5-[[4-[4-(diisobutylamino)butyl]-1-phenyl]acetyl]-10,11-dihydro 5H - -dibenzo [b,e][1,4]diazepin-11-one (DIBD). In vitro competition studies against [3H](R)-3-quinuclidinylbenzilate ([3H]QNB) and [3H]N-methylscopolamine ([3H]NMS), using membranes derived from transfected cells expressing only m1, m2, m3, or m4 receptor subtypes, indicate that DIBD is selective for m2/m4 over m1/m3. In vivo competition studies against (R,R)-[125I]IQNB indicate that DIBD crosses the blood brain barrier (BBB). The relationship of the regional percentage decrease in (R,R)-[125I]IQNB versus the percentage of each of the receptor subtypes indicates that DIBD competes more effectively in those brain regions which are known to be enriched in the m2, relative to the m1, m3, and m4, receptor subtype; however, analysis of the data using a mathematical model shows that caution is required when interpreting the in vivo results. We conclude that a suitably radiolabeled derivative of DIBD may be of potential use in emission tomographic study of changes in m2 receptors in the central nervous system. PMID- 8246671 TI - [3H] dopamine uptake by platelet storage granules in Parkinson's disease. AB - [3H] Dopamine (DA) uptake by platelet storage granules was determined in 10 never treated patients with Parkinson's disease (PD) (in 6 of them, also after 6 months of levodopa treatment), in 18 long-term levodopa-treated patients and in 15 age matched normal controls. Maximum velocity (Vmax) of DA uptake was significantly lower in the group of never-treated PD subjects compared to healthy controls (P < 0.001) and also lower compared to the PD long-term treated patients (P < 0.025). The Vmax of PD never-treated changed after submitting the patients to 6 months treatment (P < 0.025) and became similar to the Vmax in the treated group. The apparent Michaelis constant (Km) of DA uptake in PD never-treated was also significantly different from controls (P < 0.005) and also different (but not significantly) from PD "long-term" treated patients. Decreased DA uptake by platelet storage granules in PD may reflect a generalized defect of dopaminergic cells with an impairment in the vesicularization of DA which may contribute to the symptomatology of this motor disease. PMID- 8246672 TI - Differential antagonism by naltrindole-5'-isothiocyanate on [3H]DSLET and [3H]DPDPE binding to striatal slices of mice. AB - Naltrindole-5'-isothiocyanate (5'-NTII), a nonequilibrium delta opioid receptor antagonist, has been shown to antagonize differentially the antinociception induced by DSLET without affecting that induced by DPDPE. In the present study, we investigated whether or not 5'-NTII can differentially affect DSLET and DPDPE binding to receptor sites in striatal slices of mouse brain. We found that 5' NTII only changed the binding characteristics of [3H]DSLET and not that of [3H]DPDPE. Saturation binding studies revealed that 5'-NTII treatment in vitro inhibited [3H]DSLET binding by decreasing the affinity but not the number of binding sites. This finding was supported by saturation studies with [3H]DSLET in striatal slices from mice that were pretreated with 5'-NTII (10 nmol, i.c.v. or 10 mg/kg, s.c.). Thus, the results suggest that 5'-NTII can antagonize differentially the binding to striatal slices of mice of [3H]DSLET but not that of [3H]DPDPE. The binding parameters also suggest that 5'-NTII may not antagonize [3H]DSLET binding by alkylating at the receptor recognition site because the number of binding sites did not decrease. 5'-NTII may bind to some other part of the membrane to indirectly desensitize the receptor to a low affinity form. Lastly, the differential alteration of binding sites between [3H]DSLET and [3H]DPDPE by 5'-NTII strongly support the postulated existence of delta opioid receptor subtypes. PMID- 8246673 TI - In vitro progesterone effects on 3H-dopamine release from rat corpus striatum slices obtained under different endocrine conditions. AB - The release of dopamine (DA) from corpus striatum is affected by the endocrine state of the animal being progesterone suggested as a potential hormonal modulatory signal. Most of its actions have been described on endogenous DA release induced by amphetamine. However, the release of DA and the mechanism of the drug effect have been shown to be highly complexes. Considering that DA recently incorporated and/or synthetized is preferentially used we have characterized the effect of progesterone in vitro on the K(+)-induced release of 3H-dopamine (3H-DA) from rat corpus striatum slices. These were obtained during the estrous cycle or under conditions of high or low levels of endogenous progesterone (pregnant and ovariectomized rats). The release of 3H-DA was independent of the cycle. However, progesterone in vitro modified the induced release in a cycle-dependent way. Low concentrations of the hormone (100-200 mM) reduced the K+ (30 mM) effect while higher doses (300-500 mM) were facilitatories. After 7 days of ovariectomy, the induced release of 3H-DA was unchanged while in pregnant rats it was found decreased. In both cases the inhibitory effect of the hormone disappeared. Both progesterone (200 nM) and omission of Ca++ from the superfusion medium did not modified tyramine (20 microM) or K+ induced release, respectively. Data suggest that the pool of DA, related to exocytotic mechanism of release, could be specifically affected by progesterone, in a bimodal way, probably through independent genomic and non genomic influences. PMID- 8246674 TI - Glycerol-induced hyperhydration: its effects on fluid compartments in the rat. AB - Glycerol solutions are a safe, effective means to achieve a long-term state of hyperhydration. The effects of glycerol-induced hyperhydration (GIH) on the total body water (TBW), extracellular fluid (ECF), plasma volume (PV), intracellular fluid (ICF), and interstitial fluid (ISF) were investigated in the rat. Ten rats were intragastrically administered either 20 ml/kg of water or 5% glycerol. The TBW, ECF, and PV were determined by 3H2O, 14C-inulin and dye dilution, respectively at 2 h postingestion. Urine volumes and fluid retention were measured throughout the experiment. Fluid retention was increased by 50% with the 5% glycerol solution when compared to water. When the fluid compartment data at 2 h are expressed in terms of percent of body weight, the TBW and ICF of the glycerol-treated rats exceeds that of the water-treated rats, and the values for the ECF, PV, and ISF were similar after the glycerol solution and water regimens. Glycerol solutions appear to have a greater effect in expanding TBW and ICF than equal volumes of water. PMID- 8246675 TI - Melatonin binding sites are functionally coupled to phosphoinositide hydrolysis in Syrian hamster RPMI 1846 melanoma cells. AB - Recent pharmacological studies demonstrate that Syrian hamster melanoma (RPMI 1846) cells possess a nanomolar-affinity binding site for melatonin. Inhibition of 2-[125I]-iodomelatonin binding to RPMI membranes by melatonergic ligands exhibit a rank order relationship similar to that observed in hamster hypothalamus. Biochemical studies indicate that the melatonin binding site in RPMI 1846 cells is not coupled in either a stimulatory or inhibitory fashion to adenylate cyclase as a second messenger. We now report that stimulation of RPMI 1846 melanoma cells with melatonergic agonists induces phosphoinositide hydrolysis in a concentration-dependent manner (EC50: N-acetylserotonin = 0.29 microM; 2-I-Melatonin = 0.39 microM; 6-Cl-Melatonin = 0.38 microM; Melatonin = 0.45 microM). Further, phosphoinositide hydrolysis induced by 2-I-melatonin and N acetylserotonin was blocked by pre-incubation with the melatonin antagonist N acetyltryptamine and prazosin, an antagonist which exhibits potency at the nanomolar affinity melatonin binding site. 2-I-melatonin and N-acetylserotonin induced phosphoinositide hydrolysis were not blocked by the serotonin type 2 antagonist ketanserin or alpha-adrenergic antagonist, phentolamine. These data suggest that melatonin binding sites on RPMI 1846 cells are linked to phosphoinositide hydrolysis as a second messenger. PMID- 8246676 TI - The mechanism of the verapamil-digoxin interaction in renal tubular cells (LLC PK1). AB - Verapamil, usually given as a racemic mixture, decreases in vivo and in vitro digoxin renal tubular secretion, which is suggested to be mediated by P glycoprotein, an ATP-dependent multidrug efflux pump. Importantly, the two enantiomers of verapamil have been reported to similarly inhibit P-glycoprotein mediated transport of chemotherapeutic agents. In this study, we examined effects of enantiomers of verapamil on digoxin transport across an LLC-PK1 cell monolayer, a model of proximal renal tubular cells. The results indicate that verapamil inhibition of digoxin transport is non-stereospecific. Furthermore, the verapamil-digoxin interaction is not competitive. The two drugs may not share a common initial step in the P-glycoprotein-mediated transport. PMID- 8246677 TI - Kinetic and thermodynamic properties of membrane bound Ca-ATPase with low affinity to calcium in cardiac sarcolemma; response to global ischemia of the heart. AB - Thermodynamic and kinetic properties of membrane bound Ca-ATPase with low affinity to calcium in cardiac sarcolemma were investigated with respect to the effect of global ischemia on the heart. Energy barrier for ATP hydrolysis catalyzed by the Ca-ATPase was slightly higher in hearts subjected to ischemia, as it was evident from increased values of activation energy. Ischemia also induced a time dependent decrease in the activity and maximum velocity (Vmax) value of Ca-ATPase. The depression of enzyme activity was evident as early as 15 minutes after the onset of ischemia. After 30 minutes of ischemia the decrease in Vmax slowed down, probably due to an "adaptational" decrease of the Km value for Ca-ATPase. This phenomenon may be interpreted as a mechanism by which the enzyme attempts to keep working in a situation when the supply of ATP is insufficient. PMID- 8246678 TI - Circadian variations of plasma atrial natriuretic peptide and corticosterone in rats with continuous or restricted access to food. AB - In the present study plasma Atrial Natriuretic Peptide (ANP) and Corticosterone (B) levels were determined in 2 groups of Wistar rats, a Control group (C) with free access to food, and a Food Shift group (FS) with access to food only from 0900 to 1100 h for 2 weeks. Blood samples were collected by decapitation at 4 hour intervals. ANOVA indicated that B varied over time in both groups (F (6.46) = 10.14, p < 0.0001 for C and F (6.5) = 5.10, p = 0.0005 for FS). There was also time variation in plasma ANP levels in both groups (F (6.54) = 3.78, p = 0.003 for C and F (6.48) = 2.73, p = 0.02 for FS group). Plasma B presented circadian variations from 78 +/- 17 nmol/l (mean +/- SEM) at 0800 h to 339 +/- 79 nmol/l at 2000 h. The day peak plasma ANP level (pmol/l) was also attained at 2000 h (68 +/ 19). This value was higher than all others measured throughout the day. The restricted feeding regimen resulted in a 12 hour shift of peak B values (395 +/- 39 vs 125 +/- 24). The FS rats presented the highest ANP level at 0800 h. The present study indicates similar circadian variations of plasma ANP and B in rats on an ad lib or restricted feeding regimen. ANP and B secretion may occur in anticipation of the feeding period or day activity. PMID- 8246679 TI - Effects of diet-induced hyperthreoninemia. I). Amino acid levels in the central nervous system and peripheral tissues. AB - Rats were fed four levels of threonine (Thr, 0.4, 0.6, 0.8, and 5.8 g/100 g diet). After two weeks, Thr, serine (Ser), and glycine (Gly) levels were measured in plasma, liver, muscle, and central nervous system. The diet containing 5.8 g/100 g of Thr elevated Thr and Gly concentrations in plasma and nervous tissue in comparison with a standard diet. In muscle and liver, Thr concentrations were also raised but Gly levels did not change. The hepatic Thr dehydratase activity was enhanced. Diets containing moderate Thr quantities (0.6 and 0.8 g/100 g) induced slight elevations of Thr levels in all tissues. Gly concentrations were not modified. The activity of hepatic Thr dehydratase was diminished. Our results show that a high dietary content of Thr (15 times the normal levels) elevates Gly levels in various tissues, including the brain. On the contrary, diets containing 2 to 4 times the normal levels of Thr induce a weak hyperthreoninemia insufficient to modify brain Gly. PMID- 8246680 TI - Effects of ethanol ingestion on glucose transporter-1 protein and mRNA levels in rat brain. AB - In the normal adult brain, glucose provides 90% of the energy requirement, as well as substrate for nucleic acid and lipid synthesis. We have previously observed that ethanol impairs hexose uptake by rat astrocytes in culture. In the present study, male Sprague-Dawley rats, 200-250 g, were fed liquid diet in which 36% of the calories were derived from ethanol (EF) for 4 weeks. Controls were fed ad libitum (AF) or pair-fed (PF) an equicaloric diet without ethanol. Blood glucose levels did not differ between the groups at the time of study. Glucose transport by brain plasma membranes was characterized by cytochalasin B binding and showed a slight increase in transporter number (mean +/- SEM of 4 experiments = 76.4 +/- 2.5 pmoles/mg protein in EF vs. 69.5 +/- 1.0 in PF) with no change in affinity (1.8 +/- 0.1 nM-1 in EF and 1.6 +/- 0.1 in PF). Glucose transporter, GLUT-1, was increased on Western blots. In contrast, Northern analysis of cortical tissue, using a rat brain glucose transporter cDNA insert (1.59 kb Bgl II fragment of pSPGT-1), showed a 23 to 35% decrease in steady-state levels of glucose transporter mRNA. GLUT-1 mRNA, localized in brain sections by in situ hybridization histochemistry, showed marked reductions in choroid plexus and hippocampus following ethanol treatment. Ethanol appears to have multiple effects on brain GLUT-1. PMID- 8246681 TI - Cognition stimulating drugs modulate protein kinase C activity in cerebral cortex and hippocampus of adult rats. AB - The in vivo and in vitro effect of oxiracetam, aniracetam and alpha glycerylphosphorylcholine (alpha GPC) on protein kinase C (PKC) activity was studied in rat brain cortex and hippocampus. Administration of oxiracetam and alpha GPC in vivo elicited an early increase of particulate histone-directed PKC activity accompanied by a decrease of soluble activity and followed a few hours later by a down regulation of the enzyme. The effect was also observed in vitro when either oxiracetam or alpha GPC were administered at nanomolar concentrations to rat brain cortex slices. Aniracetam had no effect in the cortex but promoted PKC translocation both in vivo and in vitro in the hippocampus. In cortex slices the effect of oxiracetam was antagonized by the addition of AP-5, an NMDA receptor blocker, but not by CNQX and L-AP3, antagonists of AMPA and metabotropic glutamate receptors, respectively. Scopolamine also prevented the increase of particulate PKC elicited by oxiracetam in vitro. In the hippocampus the increase of particulate PKC activity was antagonized by AP-5, CNQX and L-AP3, indicating participation by both ionotropic and metabotropic glutamate receptors in the action of aniracetam. The data support the hypothesis that PKC activation may be a common mechanism amongst cognition stimulating drugs from different chemical classes. PMID- 8246682 TI - The indirect pathway of hepatic glycogen synthesis and reduction of food intake by metabolic inhibitors. AB - The increasingly recognized role of the indirect pathway (glycolysis followed by hepatic gluconeogenesis) for glucose utilization and glycogen synthesis by the liver led us to administer 3-mercaptopicolinate (3MP), an inhibitor of phosphoenolpyruvate-carboxykinase, in an attempt to assess the role of liver glycogen or hexose-phosphates in the food-intake reducing effects of (-)hydroxy citrate. Administration of (-)hydroxy-citrate increased hepatic glycogen content in i.v. glucose refed rats. Using the glucuronide probe technique, the mechanism of increased glycogen deposition was shown to be prolongation of indirect pathway (recycled) input. Daily (-)hydroxy-citrate significantly reduced food intake (from 12.0 +/- 2.3 to 6.4 +/- 3.6 g/day, p < 0.05) and had no chronic effect on hepatic glycogen content in rats trained to a single daily meal (meal-fed). Administration of 3MP completely suppressed hepatic glycogen synthesis (< 0.5 mg/g) when given alone or with (-)hydroxy-citrate. Isotopic studies confirmed inhibition of the indirect pathway of UDP-glucose synthesis. 3MP accentuated rather than prevented the (-)hydroxy-citrate reduction in food intake in meal-fed rats (intake 2.7 +/- 2.4 g/day). When given alone, 3MP also reduced intake (6.1 +/- 3.6 g/day). Severe hypoglycemia was observed (glucose < 20 mg/dl) in several meal-fed rats given repeated daily doses of 3MP, yet food intake did not occur despite food availability. Neither 3MP nor (-)hydroxy-citrate had any effects when given after the daily meal. We conclude that the role of the indirect glycogen synthesis pathway must be considered in any theory of regulation of food intake by hepatic metabolites and that, if the effects of these metabolic inhibitors can be shown not to be toxic or non-specific, neither hepatic glycogen nor hexose-phosphates are involved in the food-intake suppressive effects of ( )hydroxycitrate. PMID- 8246683 TI - Relationship between cyclic-AMP content, regional myocardial function and O2 consumption in experimental left ventricular hypertrophy: effect of negative inotropes. AB - The aim of this study was to examine the hypothesis that negative inotropic agents that lower myocyte cyclic-AMP by different means would have similar effects on local myocardial segment work and O2 consumption in control hearts, but that this response would differ in left ventricular hypertrophy (LVH) induced by aortic valve stenosis. Open chest anesthesized LVH and control dogs were studied before and during esmolol (100 micrograms/kg/min) and acetylcholine (100 micrograms/kg/min) infusion. Regional work was calculated as the integrated product of instantaneous force (miniature transducer) and shortening (sonomicrometry) per min. Regional O2 consumption was calculated from blood flow (radioactive microspheres) and O2 saturation of small frozen vessels (microspectrophotometry). Cyclic-AMP level was determined with a competitive binding assay using 3H-cyclic-AMP and was found to be 731 +/- 90 (mean +/- S.D.) pmol/g in control and 711 +/- 163 in LVH. There were similar decreases in cyclic AMP levels in control hearts with acetylcholine (365 +/- 135) and the beta adrenergic blocker (430 +/- 95). In LVH, esmolol lowered cyclic-AMP (383 +/- 39), but acetylcholine did not (689 +/- 105). In control animals, regional O2 consumption (7.7 +/- 0.6, 5.6 +/- 0.4 and 5.6 +/- 0.5 ml O2/min/100 g, control, acetylcholine, esmolol, respectively) and segment work (878 +/- 82, 546 +/- 80, 627 +/- 66 g*mm/min) fell to similar levels with these agents. Similar decreases were found in LVH with esmolol for O2 consumption (7.1 +/- 1.2, 5.1 +/- 1.0, baseline, esmolol) and segment work (895 +/- 140, 427 +/- 65). Acetylcholine had no significant effect on segment work (800 +/- 201), but did lower regional O2 consumption (4.0 +/- 0.7) in LVH dogs. It is concluded that there is a strong relationship between the level of cyclic-AMP and myocardial function and O2 consumption in control hearts. The action of acetylcholine is altered in LVH leading to an uncoupling between regional cyclic-AMP, function and metabolism. PMID- 8246684 TI - Bombesin stimulates intracellular Ca2+ mobilization but not proliferation on human colon cancer cells. AB - Increases in intracellular Ca2+ ([Ca2+]i) levels mediated by bombesin (BBS) are believed to be important signals leading to stimulation of DNA synthesis and an increase in cellular proliferative rate. Since the role of BBS on growth of normal or malignant cells in the GI tract is still unclear, we examined whether BBS affects in vitro growth of human colon cancer cells (COLO 320, HCT116 and LoVo). We also examined the effect of BBS on intracellular Ca2+ levels to determine if the growth-regulatory effect of BBS is mediated through increases in [Ca2+]i. Levels of [Ca2+]i in response to BBS were measured by single cell fluorescence after loading with fura-2. BBS stimulated the mobilization of [Ca2+]i in COLO 320, LoVo, and HCT116 cells in a dose-dependent fashion, but did not affect in vitro growth. These findings suggest that the BBS-mediated increase in [Ca2+]i does not always correlate with the growth-regulatory effect of BBS. PMID- 8246685 TI - Modulation by sphingosine of substrate phosphorylation by protein kinase C in bovine mammary gland. AB - The effect of sphingosine on the phosphorylation of endogenous proteins by protein kinase C (PKC) was investigated in bovine mammary gland. Several proteins were shown to be substrates for PKC in both cytosolic and total particulate fractions by phosphorylation in the absence or presence of 1-oleoyl-2-acetyl-sn glycerol, phosphatidylserine (PS) and Ca2+. At concentrations of 83 microM or less, sphingosine inhibited phosphorylation of several substrates for PKC in both fractions. Phosphorylation of cytosolic 36 kDa, 21 kDa and particulate 36 kDa proteins was particularly sensitive to sphingosine. Cytosolic 97 kDa phosphorylation (which was enhanced by Ca2+ alone) was also sensitive to sphingosine. The inhibition was reversed by excess addition of lipid cofactors, particularly PS, but not by Ca2+. At higher concentrations (167 and 417 microM), in addition to the inhibition seen at lower concentrations, sphingosine stimulated phosphorylation of several proteins, including cytosolic 19 kDa and particulate 53 kDa, which were not detected in the absence of sphingosine. The sphingosine-induced phosphorylation disappeared with excess addition of PS, but not with addition of Ca2+. The results point toward the importance of the interaction of sphingosine with membrane phospholipids in the signal transduction pathway mediated by PKC-dependent phosphorylation in bovine mammary gland. PMID- 8246686 TI - Intravenous injection of tridihomo-gamma-linolenoyl-glycerol into mice and its effects on delayed-type hypersensitivity. AB - Highly purified tridihomo-gamma-linolenoyl-glycerol (DGLA-TG) was emulsified with egg yolk lecithin as a 10% (wt/vol) DGLA-TG emulsion. We injected 0.05 or 0.5 mL of the emulsion into mice through the tail vein and investigated its effects on the fatty acid composition of spleen cells and on delayed-type hypersensitivity (DTH) response. At 1 h after the injection, dihomo-gamma-linolenic acid (DGLA) concentrations were increased significantly in the total phospholipid fraction of spleen cells from 1.21 +/- 0.13 mol% to 2.09 +/- 0.74 mol% (P < 0.02) and 7.95 +/ 1.25 mol% (P < 0.001) in the 0.05-mL and 0.5-mL groups, respectively. Mice, which had already been immunized with sheep red blood cells (SRBC), were challenged by the injection of SRBC into the right-hind footpad. Intravenous injection into mice with 0.5 mL of the emulsion immediately before the challenge almost completely suppressed DTH response measured by the swelling of the right hind footpads 24 h thereafter. This inhibitory effect on the DTH response was significant with as little as 0.05 mL of the emulsion, whereas a soybean oil emulsion was not effective at all. In conclusion, intravenous injection of a DGLA emulsion increased DGLA concentrations in immune cells within 1 h and suppressed the DTH reaction. PMID- 8246687 TI - pH-dissociation characteristics of cardiolipin and its 2'-deoxy analogue. AB - Cardiolipin (CL) is found in inner mitochondrial membranes and the plasma membrane of aerobic prokaryotes. CL is tightly bound to those transmembrane enzymes associated with oxidative phosphorylation. CL has earlier been reported to have a single pK at low pH. We have titrated CL in aqueous suspension (bilayers) and in solution in methanol/water (1:1, vol/vol) and found it to display two different pK values, pK1 at 2.8 and pK2 initially at 7.5 but shifting upwards to 9.5 as the titration proceeds. The unusually high pK2 might be explained by the formation of a unique hydrogen bond in which the free hydroxyl on the central glycerol forms a cyclic intramolecular hydrogen-bonded structure with one protonated phosphate (P-OH group). We have therefore chemically synthesized the 2'-deoxycardiolipin analogue, which lacks the central free hydroxyl group, and measured its pH-dissociation behavior by potentiometric titration, under the same conditions as those for CL. The absence of the hydroxyl group changes the titration dramatically so that the deoxy analogue displays two closely spaced low pK values (pK1 = 1.8; pK2 = 4.0). The anomalous titration behavior of the second dissociation constant of CL may be attributed to the participation of the central glycerol OH group in stabilizing the formation of a cyclic hydrogen-bonded monoprotonated form of CL, which may function as a reservoir of protons at relatively high pH. This function may have an important bearing on proton pumping in biological membranes. PMID- 8246688 TI - Modulating effects of bile salt hydrophobicity on bile secretion of the major protein of the bile lipoprotein complex. AB - Bile lipids are secreted in association with a newly identified major apoprotein called anionic polypeptide fraction-calcium binding protein (APF-CBP), which is synthesized in the hepatocytes and has been detected in both bile and plasma and characterized. The secretion of the lipids in bile depends both on the concentration and the hydrophobicity of the bile salts (BS) secreted. The present study was undertaken to determine whether the synthesis and the secretion of APF CBP are similarly regulated by BS, using two methods. The synthesis and secretion of labelled, newly synthesized APF-CBP by isolated rat hepatocytes were monitored by solid-phase immunoassay. For this purpose, hepatocytes were incubated with either glycodeoxycholate (GDC) or taurocholate (TC). The synthesis and secretion of labelled, newly synthesized APF-CBP by perfused rat liver were measured by immunological enzyme-linked assay (ELISA) upon perfusing the liver with either GDC or TC. We found that (i) the synthesis and the secretion of APF-CBP were increased during either TC or GDC perfusion, but the increase was more pronounced with TC; (ii) in GDC perfusion the APF-CBP levels measured were more closely related to the levels of bile salts and not to phospholipid levels, (iii) when the two bile salts were perfused in reverse order, i.e., first GDC and then TC, the secretion of APF-CBP in bile decreased when GDC was perfused, but increased when TC was perfused. Similar results were obtained in experiments with isolated hepatocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246689 TI - 3-Carboxy-4-nitrophenyl-dithio-1,1',2-trisnorsqualene: a site-directed inactivator of yeast oxidosqualene cyclase. AB - The role and location of essential thiol groups in 2,3-oxidosqualene cyclase from Saccharomyces cerevisiae was examined (i) by comparing inactivation properties of two known thiol reagents, 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and 2-nitro 5-thiocyanobenzoic acid (NTCB), with 3-carboxy-4-nitrophenyl-dithio-1,1',2 trisnorsqualene (CNDT-squalene), a new thiol reagent designed as a site-directed inactivator of oxidosqualene cyclase and (ii) by testing the ability of the substrate to protect the enzyme against inactivation by the reagents. All reagents gave a time-dependent inactivation following pseudo-first order kinetics. DTNB and CNDT-squalene showed comparable inactivation ability (Ki = 0.67 and 1.21 mM), whereas NTCB was less effective (Ki = 15.6 mM). Strong differences between the two most active inhibitors, DTNB and CNDT-squalene, were observed when the enzyme was saturated with substrate prior to incubation with the thiol reagent. While substrate did not protect the enzyme against the inactivation caused by DTNB, a reduction in the inactivation ability of CNDT squalene was observed under protection conditions. The data suggest that the squalene-like inactivator modifies a thiol group located at the active site of the enzyme. PMID- 8246690 TI - General resistance to sterol biosynthesis inhibitors in Saccharomyces cerevisiae. AB - Screening for resistance to fenpropimorph was undertaken in order to isolate yeast mutants affected in the regulation of the ergosterol pathway. Among the mutants isolated, one bearing the recessive fen1-1 mutation was characterized by a 1.5-fold increase in the ergosterol level and a general resistance to sterol biosynthesis inhibitors. The fen1-1 mutation was linked to MAT locus on chromosome III. The measurement of enzyme activities involved in the ergosterol pathway revealed that isopentenyl diphosphate (IPP) isomerase activity was specifically increased 1.5-fold as compared to the wild type strain. However, overexpression of IPP isomerase in the wild type strain was not by itself sufficient to lead to sterol increase or resistance to sterol biosynthesis inhibitors, showing that IPP isomerase is not a limiting step in the pathway. The fen1-1 mutation permits viability in aerobiosis of yeast disrupted for sterol-14 reductase in absence of exogenous ergosterol supplementation, whereas the corresponding strain bearing the wild type FEN1 allele grows only in anaerobiosis. This result shows that ignosterol is able to efficiently replace ergosterol as bulk membrane component and that the fen1-1 mutation eliminates the specific ergosterol requirement in yeast. PMID- 8246691 TI - Effect of lipid content of diet on cholesterol content and cholesterogenic enzymes of European eel liver. AB - The effect of dietary lipid levels on the levels of cholesterol and the activities of the major cholesterogenic enzymes of the liver has been studied in the European eel. An increase in hepatic total cholesterol was observed when the dietary lipid levels increased from 12 to 20%, while protein levels were maintained at 30%. This change paralleled an increase in mevalonate 5 pyrophosphate decarboxylase activity, while 3-hydroxy-3-methylglutaryl-CoA reductase mevalonate kinase and mevalonate 5-phosphate kinase were not affected by changes in diet composition. These results suggest that the decarboxylase may be a rate-limiting enzyme in cholesterogenesis in eel liver. PMID- 8246692 TI - Parameters influencing cholesterol oxidation. AB - The purpose of this study was to investigate the effects of temperature, oxidation time, presence of water, pH, type of buffer and form of substrate used on cholesterol oxidation. Microcrystalline cholesterol films, both solid and melted, and aqueous suspensions of film fragments were used as substrates. Use of dispersing agents was avoided. Quantitative analysis of the unaltered substrate and the products of its autoxidation was carried out by gas chromatography over the course of oxidation. Solid cholesterol films were found to be resistant to autoxidation in the dry state. However, when heated at 125 degrees C, a sudden increase in oxidation rate occurred at a point coinciding with the visible melting followed by a plateau of the oxidation rate. All of the autoxidation products formed underwent further decomposition. Film fragments of cholesterol oxidized at a faster rate in aqueous suspensions than when oxidized in the dry state. In aqueous suspensions, the differences in the resistance of cholesterol to oxidation were not significant within the pH range 6.0-7.4, except for the early stages of oxidation. The 7-ketocholesterol/7-hydroxycholesterol ratio dropped significantly with increasing pH. However, at all pH levels tested, this ratio remained relatively constant during the 6 h of heating. While the 7 beta hydroxycholesterol/7 alpha-hydroxycholesterol ratio was not affected by pH in the range of 6.0-7.4, at pH 7.4 a high preference was observed for the cholesterol beta-epoxide over its alpha-isomer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246693 TI - Cholesterol and 25-hydroxycholesterol retention in specimens of liver and aorta prepared for electron microscopy. I. Standard fixation methods and metabolism of the labeled sterols. AB - In the present work, several preparatory procedures commonly used for electron microscopy (EM) were evaluated as to their ability to preserve cholesterol (CHO) and CHO derivatives in tissue. We also determined in several rat tissues to what extent the sterols used as tracers are metabolized. Sprague-Dawley rats were injected intraperitoneally with [1 alpha,2 alpha(n)-3H]cholesterol ([3H]CHO) and 25-hydroxy-[26,27-3H]cholesterol ([3H]25-OH-CHO). Lipids of the liver, aorta and brain were extracted one and five days after injection, and the distribution of the labeled lipids was followed by thin-layer chromatography. When labeled CHO was injected as tracer, most of the radioactivity remained associated with the CHO fraction. When 25-hydroxycholesterol (25-OH-CHO) was used, we found that it was mostly metabolized to yield more polar compounds. Our results show that the loss of CHO and CHO derivatives from tissues depends not only on the preparatory procedure used for EM, but also on the type of tissue studied. PMID- 8246694 TI - Cholesterol and 25-hydroxycholesterol retention in specimens of liver and aorta prepared for electron microscopy. II. Effect of filipin, osmium, digitonin and saponin. AB - Sprague-Dawley rats were injected intraperitoneally with [1 alpha, 2 alpha(n) 3H]cholesterol or 25-hydroxy-[26,27-3H]cholesterol, and one and five days later liver and aortic tissues were fixed. The extent to which these sterols were lost from the tissues during preparation for electron microscopy (EM) was examined utilizing different fixation procedures and various protective agents. Radioactive tracers, scintillation counting and standard EM techniques were used. Although most of the procedures examined caused major lipid losses, useful fixation procedures that allow retention of cholesterol or 25-hydroxy-cholesterol in liver and aortic tissues were found and are described here. PMID- 8246695 TI - A micro enzymic method for determination of choline-containing phospholipids in serum and high density lipoproteins. AB - A micro method is described for the assay of choline-containing phospholipids in serum and high density lipoproteins (HDL) using an automated microtiter plate reader. The method is adapted from the enzymic method of Takayama, Itoh, Nagasaki, and Tanimuzu (Clin. Chim. Acta 79, 93-98, 1977) using phospholipase D, choline oxidase, and peroxidase coupled with the color generating system phenol and 4-amino-antipyrine. The micro method requires 5 microL of serum or HDL sample, and 42 samples can be assayed in duplicate in one run using a 96-well flat-bottom microtiter plate. The reaction is linear up to 400 mg/dL and the lower limit of detection is 0.25 mg of choline-containing phospholipids per assay. The coefficient of variation within an assay is 0.86-0.79%, and day-to-day variation is 0.9-1.5%. Results obtained by the micro method are in excellent agreement with those obtained by the procedure of Takayama et al. (r = 0.997). The supernatant left after removal of low density lipoproteins and very low density lipoproteins from serum and precipitation with heparin/manganese chloride reagent can thus be conveniently used for the micro assay of choline phospholipids in HDL. PMID- 8246696 TI - Comments on erythrocyte membrane phospholipid fatty acid changes in cerebral palsy patients during nutritional rehabilitation. PMID- 8246697 TI - QALYs, HYEs, and the loss of innocence. PMID- 8246698 TI - A neural-network approach to predicting admission decisions in a psychiatric emergency room. AB - Clinical decision making is based on recognizing complex patterns of patients' signs and symptoms. Neural networks have been shown to be very effective at this type of pattern recognition, and in this study a neural-network approach was used to predict which patients seen in a psychiatric emergency room required admission and which did not. Data from all walk-in patients (N = 658) evaluated during normal working hours in a psychiatric emergency room during a one-year period were used either to train a neural network or to test its performance. The network had 53 input nodes, one hidden layer, and an output layer with a single node. The back-propagation method was used to train the network. The neural network's admitting decisions were in substantial agreement with those of the clinicians (kappa coefficient = 0.63). When used as a diagnostic test for admission it had a specificity of 94%, a sensitivity of 70%, and an overall accuracy of 91%. The information gain was 35% of that of a perfect diagnostic test. These results show that a neural network can be trained to make clinical decisions that are in substantial agreement with those of experienced clinicians. PMID- 8246699 TI - Are healthy-years equivalents an improvement over quality-adjusted life years? AB - The construct of the healthy-years equivalent (HYE) has been proposed as an alternative to the quality-adjusted life year (QALY) on the grounds that it avoids certain restrictive assumptions about preferences and also incorporates attitudes toward risk. The authors review the construct of the QALY, including both the commonly used risk-neutral formulation and the more general formulation that permits risk aversion (or risk preference) with respect to remaining life years. They show that the HYE adds flexibility to the risk-neutral form of the QALY by permitting the rate of tradeoff between life years and quality of life to depend on the life span, albeit at the cost of eliciting numerous additional time tradeoff assessments. However, the claim that the HYE incorporates attitudes toward risk is incorrect, and the proposed two-stage procedure to measure HYEs is neither necessary nor sufficient to incorporate attitudes toward risk. In fact, the HYE assumes risk neutrality with respect to healthy years of life and, therefore, is less suitable for decisions under uncertainty than is the general (risk-averse) form of the QALY. PMID- 8246700 TI - Healthy-years equivalents versus quality-adjusted life years: in pursuit of progress. AB - The authors respond to the contention that healthy-years equivalents (HYEs) do not fulfil claims they previously made. They refute the argument for the equivalence of the two-stage lottery and the time tradeoff and show that it fails to recognize the difference between choice problems under uncertainty versus certainty. They also dismiss the claim that HYEs assume risk neutrality with respect to healthy years of life, and show that HYEs are superior to the "general" (risk-averse) form of QALYs because HYEs employ fewer assumptions; in particular, they do not assume a specific form of the individual's utility function. The examples they used earlier are demonstrated to be conclusive, and the authors respond to arguments about the practical aspects of using HYEs vs QALYs. Finally, they take issue with the assumption that the roots of the QALY model are all attributable to utility theory. PMID- 8246701 TI - Practice variation in the management of pharyngitis: the importance of variability in patients' clinical characteristics and in physicians' responses to them. AB - The objective of this study was to assess whether geographic differences in antibiotic-prescribing rates for patients with pharyngitis could be explained by intersite differences in patients' clinical characteristics and in how physicians responded to these clinical cues when making decisions. As part of the initial phase of a prospective controlled trial to improve physicians' diagnostic ability, the authors enrolled cohorts of consecutive patients seen at staff-model -HMO student health services in Pennsylvania and Nebraska. Physicians' decisions whether to prescribe antibiotics for 310 consecutive patients presenting with pharyngitis to the former and 214 such patients presenting to the latter at the time of the initial visit were examined. There was a large discrepancy between the antibiotic-prescribing rates at the student health services in Pennsylvania, 106/310, 32.4%, and Nebraska, 156/214, 72.9%. The clinical variables significantly independently associated with treatment at both sites in a logistic regression model were fever, adjusted odds ratio = 2.1 (95% CI = 1.1, 3.8); exudates, 5.4 (2.8, 10); palatine petechiae, 6.5 (1.5, 28); rhinorrhea, 0.46, (0.25, 0.85); and high risk of complications, 3.8 (1.04, 14). There was a significant interaction between site and anterior cervical adenopathy, 5.5 (1.6, 19); and a borderline interaction between site and rhinorrhea, 2.4 (0.89, 6.7). Site was not a significant independent predictor of treatment, 1.8 (0.45, 6.6.). Practice variation was related to geographic differences in patients' clinical characteristics and in how physicians responded to these factors when prescribing antibiotics. How physicians weight patients' clinical characteristics when making decisions may be an important element of their "practice styles." PMID- 8246702 TI - Contingent valuation of supplemental health care in Israel. AB - The use of the contingent-valuation method for determining willingness to pay for non-market or currently available health care services continues to be experimental. In this study, the contingent-valuation method was used to calculate willingness-to-pay estimates for a proposed change in the Israeli health care system. It was found that the willingness-to-pay estimates calculated in the Israel study were reasonable and that the methodology is able to adapt to the special nature of the health care commodity while adhering to the conditions for reliability and validity in a contingent-valuation study. PMID- 8246703 TI - The contingent-valuation method. PMID- 8246704 TI - Estimating diagnostic accuracy from multiple conflicting reports: a new meta analytic method. AB - Reports of diagnostic accuracy often differ. The authors present a method to summarize disparate reports that uses a logistic transformation and linear regression to produce a summary receiver operating characteristic curve. The curve is useful for summarizing a body of diagnostic accuracy literature, comparing technologies, detecting outliers, and finding the optimum operating point of the test. Examples from clinical chemistry and diagnostic radiology are provided. By extending the logic of meta-analysis to diagnostic testing, the method provides a new tool for technology assessment. PMID- 8246705 TI - Markov models in medical decision making: a practical guide. AB - Markov models are useful when a decision problem involves risk that is continuous over time, when the timing of events is important, and when important events may happen more than once. Representing such clinical settings with conventional decision trees is difficult and may require unrealistic simplifying assumptions. Markov models assume that a patient is always in one of a finite number of discrete health states, called Markov states. All events are represented as transitions from one state to another. A Markov model may be evaluated by matrix algebra, as a cohort simulation, or as a Monte Carlo simulation. A newer representation of Markov models, the Markov-cycle tree, uses a tree representation of clinical events and may be evaluated either as a cohort simulation or as a Monte Carlo simulation. The ability of the Markov model to represent repetitive events and the time dependence of both probabilities and utilities allows for more accurate representation of clinical settings that involve these issues. PMID- 8246706 TI - Collaborative medical decision making. AB - Collaborative decision making occurs whenever two or more individuals contribute their diverse knowledge and expertise to the decision-making process. In medicine, this happens during morning rounds, case conferences, consultations, and elsewhere. This paper presents an analysis of collaborative medical decision making, focusing on two factors that can powerfully influence the kind of information that gets discussed, and hence the nature of the decisions that are made. These are 1) the pre-discussion distribution of problem-relevant information/knowledge, and 2) each participant's awareness of other individuals' knowledge and talents. The authors review previous psychological research on group decision making that concerns these factors, and call attention to several lines of inquiry that might fruitfully be pursued in clinical settings. PMID- 8246707 TI - Simulation modeling and health-care decision making. PMID- 8246708 TI - Graphic representation of numerically calculated predictive values: an easily comprehended method of evaluating diagnostic tests. AB - The authors present a brief BASIC-language computer program that calculates positive and negative predictive values from the known sensitivity and specificity of a test over a range of disease prevalences and produces easily comprehended graphs of these relationships using commonly employed graphics software. These graphs permit rapid appreciation of the effect of prior probability on predictive values. They may facilitate selection of the appropriate test for the anticipated setting and avoidance of the unexpected inaccuracies that often result when tests are chosen on the basis of sensitivity and specificity alone. PMID- 8246709 TI - Twelve tips for using videotape reviews for feedback on clinical performance. PMID- 8246710 TI - Teaching and assessment in the consultation. A workshop for general practice clinical teachers. AB - A faculty development programme for general practice clinical teachers offered a skills development workshop which specifically addressed the skills needed when teaching and assessing students consulting with patients. The course was sequential, that is, a group moved on to a new skill only when its members could demonstrate mastery of a lower-order skill. Participants practised skills and received feedback on their performance from peers and learners (students and trainees). Participants comments about the course were highly favourable and the majority foresaw a general improvement in their own personal consultation skills as a result. They recognized that the use of a valid and reliable set of criteria (the Leicester Assessment Package) was essential in order to make correct judgements about consultation performance before instituting remedial teaching. PMID- 8246711 TI - A proposal for enhancing the quality of clinical teaching: results of a department of medicine's educational retreat. AB - New challenges for clinical teachers include incorporating a deeper appreciation of the use of the published literature in day-to-day practice and teaching, responding to the profusion of diagnostic tests and treatments, and dealing with changing practical difficulties. We report a summary of a retreat conducted by our Department of Medicine in which our faculty presented and refined strategies they had developed to deal with these challenges. Areas of discussion include developing an effective medical team managing time on a busy clinical service, teaching pathophysiology, teaching clinical skills, and teaching critical appraisal. Our observations are likely to be useful to clinicians involved in patient-centred teaching in wards and clinics, particularly those interacting with groups of undergraduate and post-graduate trainees. PMID- 8246712 TI - Implications of adult education theories for medical school faculty development programmes. AB - Adult education principles have impacted the approach used by many faculty development programmes. Underlying themes of immediacy of application, task centered opportunities to practice skills, and feedback on efforts are common to most popular approaches. Less common are instances where the need to know is initially communicated, or provision of opportunities to direct course content. Incorporation of sound adult educational principles into the design of faculty development programmes should enhance its reception by the faculty, and increase its value to the school. PMID- 8246713 TI - Quality assurance in graduate medical education: a peer review process. AB - The Accreditation Council for Graduate Medical Education in the United States has recently required institutions with residency and fellowship training programmes to develop an Institutional Policy Review Document outlining how the educational quality of these programmes is monitored. Whereas departments may have assumed this responsibility in the past, this is clearly a function of the institution. The purpose of this paper is to describe a long-standing, effective, internal peer review process to assure quality education in an institution. PMID- 8246714 TI - Physicians know how to be good educators too: a comparison of the patient problem solving process and the instructional design process. AB - Education should be viewed as one of the important roles of a physician. Yet, because physicians generally are not trained to be educators, many instructional sessions, lectures and workshop presentations are ineffective because the instructional design process has been ignored. While not a substitute for professional education courses, physicians can use knowledge of the patient problem solving process to develop effective educational interventions. Both the patient problem solving process and the instructional design process consist of seven nearly identical steps which follow a basic scientific problem-solving approach. Knowledge of this similarity can improve the effectiveness of educational activities developed by physicians. PMID- 8246715 TI - A model to teach trauma care to medical students. AB - The following integrated trauma teaching model has been developed in response to perceived deficiencies of organized trauma content in many undergraduate surgical programmes. This method has combined structured teaching and clinical clerkship. First, a short lecture is given to the student body giving an overview of the subject. Small groups then rotate through a series of teaching stations, each structured to represent a different level in the hierarchy of management priorities involved in the care of a trauma patient. The educational value of a 150 minute teaching session was evaluated by comparing group performance in a pre test given several days before with a post-test immediately after teaching. Forty one Year IV students increased their mark, out of a possible total of 30, from a pre-test mean 15 (SD3) to a post-test mean 21 (SD3). Thirty two year VI students increased their marks from a mean 18 (SD3) to a mean 24 (SD2). Both increases are highly significant (p < 0.001). The model has proved to be an effective way to teach students trauma skills in preparation for a subsequent clerkship or internship. PMID- 8246716 TI - A programme for developing clinical reasoning skills in graduate physiotherapists. AB - This paper presents a clinical reasoning course for postgraduate physiotherapy students, conducted at the University of Sydney. The course was designed to foster the development of students' clinical reasoning skills and clinical knowledge. The rationale and nature of the course are discussed, along with evaluation findings. Results indicate that the course has succeeded in stimulating students' interest and enjoyment in clinical reasoning and promoting students' understanding of and competence in this complex ability. PMID- 8246717 TI - The ongoing development of a problem-based surgery clerkship: year three. AB - The third-year clerkship at one college of medicine was modified to reflect an emphasis on adult learning principles and life-long learning habits. Problem based learning was selected as the educational method, and the development of a cognitive knowledge base, clinical problem-solving skills, and appropriate clinical skills was stressed. At a yearly faculty education retreat, strengths and weaknesses of the clerkship were discussed. As a result of this meeting, several changes were made in the clerkship, including additional stress on students' usage of the medical library and computerized databases, a tutor development program, the addition of expert resource sessions, and more emphasis on students' development of clinical skills through a preceptor program. A comprehensive evaluation program designed to support the educational goals of the clerkship is in place. The major challenge of the program remains the proper synchronization of curricular, clinical, and student self-study activities essential for an ideal learning experience. PMID- 8246718 TI - Attendings' and residents' teaching role and students' overall rating of clinical clerkships. AB - The study was conducted with a sample of third-year students (n = 584) at Jefferson Medical College to explore students' perception of patterns of differences between attending physicians and residents in their teaching behaviors during clinical clerkships. Attending physicians' teaching behaviors were perceived more in a mentorship mode whereas residents' teaching behaviors were equally divided between mentorship and preceptorship modes. Attending physicians and residents' teaching behaviors varied among clerkships. Results were discussed in terms of difference of teaching roles played by attending physicians and residents and relationship of the teaching behaviors to students' overall rating of clerkship. PMID- 8246719 TI - The structured oral self-directed learning evaluation: one method of evaluating the clinical reasoning skills of occupational therapy and physiotherapy students. AB - The Structured Oral Self-directed Learning Examination (SOSLE) is used to evaluate the clinical reasoning skills of occupational therapy (OT) and physiotherapy (PT) students. It is an oral examination which evaluates a student's problem-solving ability, self-directed learning skills, knowledge level and self-assessment ability. The three parts of the examination are conducted over a 24-hour period. Validation of this instrument was carried out in two groups of OT and PT undergraduate students over two consecutive years (Year 1--n = 20) (Year 2--n = 18). Inter-rater reliability correlations varied from 0.61 to 0.78 the first year to 0.85 to 0.99 in the second year. The results obtained from the SOSLE were also compared to written and tutorial marks obtained in the same course. Pearson Correlation Coefficients (PCC) among mean SOSLE and two written paper scores ranged from 0.0-0.05 (Year 1) to 0.0-0.1 (Year 2). The PCC among the mean SOSLE and tutorial performance scores were 0.57 (Year 1) and 0.0 (Year 2). The results show that good agreement between raters can be reached using this evaluation method. However, the poor correlations between the SOSLE and the other methods of evaluation may show that different skills are being evaluated. Further validity testing needs to be carried out to confirm that this tool is measuring process oriented skills. PMID- 8246720 TI - A reappraisal of the use of multiple choice questions. AB - The terms Multiple True/False and Best Answer deserve wider applicability than at present when discussing multiple choice questions (MCQs). They describe the truthfulness of the choices and distinguish between the question types. The choice which matches the key in a MCQ is best called the correct answer. Multiple True/False questions contain either true (accepted) or false (unaccepted) ideas in the correct answers while in the Best Answer type, all choices are true. Using a complex recording procedure to indicate the selections already made, as in the K-type MCQ, does not assess any additional abilities. Candidates answering Multiple True/False MCQs should be asked to respond to each choice as either true or false. This helps to discriminate between those who can detect the falsity of a false choice from those who cannot. When dealing with guessing, examiners need to be cognizant of issues such as blind guessing and informed guessing, proclivity for risk-taking, extension of the marking range and probabilities according to the binomial distribution. PMID- 8246721 TI - Could final year school grades suffice as a predictor for future performance? AB - The performance of three groups of medical students was evaluated at the end of one phase of their studies--after 2 years of premedicine. The aim of the study was to determine whether any of the assessment methods--secondary school grades, entrance examination or test for proficiency in the English language, used for selection of medical students, could reliably predict student performance in the early stages of their education. A significant correlation was found between secondary school grades and the entrance exam on the one hand and grade point average (GPA) on the other. However the results of the test for English proficiency did not correlate with the GPA. It is concluded that the secondary school grade on its own is a good predictor of academic performance in the early stages of medical undergraduate education. PMID- 8246722 TI - What's new in education: items of interest to medical educators. PMID- 8246723 TI - Do you know? An opportunity to assess how up-to-date you are with the medical education literature. PMID- 8246724 TI - [Serological, clinical, and epidemiological aspects of an outbreak of trichinosis in Azul, province of Buenos Aires]. AB - An outbreak of human trichinellosis that took place in Southern Buenos Aires Province, an endemic area for this zoonosis, was studied. Eighteen individuals were infected by eating swine sausages lacking veterinary control. Clinical symptoms, as well as serological studies related to antibody production and cytotoxic capacity of the sera are described. Eight interdicted pigs and their products were analysed. In order to determine the transmission network of this zoonosis, wild rats were hunted in the farm where the swine were bred. The clinical symptoms of the infected individuals were: myalgia, edema, fever and cephalea. Sixty-one percent of the patients showed antibodies against muscle larvae 15-20 days after ingestion of the infected meat. Five out of eighteen patients showed anti-newborn larvae antibodies 40-60 days postinfection. The sausage burden was 5.3 muscle larvae/g. Two out of eight swine were diagnosed employing serological techniques, while parasitic techniques detected only one. The rat is an important reservoir in the domestic cycle of this zoonosis. PMID- 8246725 TI - Evaluation of the blood components considered as risk predictors in coronary heart disease. AB - The study included 249 patients two days before cardiovascular surgery and 73,915 control subjects. Results obtained were analyzed by grouping the individuals according to sex and age. In coronary heart disease (CHD) in males, total cholesterol was found higher than in controls (mean +/- D.S.: 241.9 +/- 44.7 vs 223.6 +/- 43.0 mg/dl, p < 0.01) between 25 and 49 years of age, this significance being lost with age. Triglycerides were also higher (197 +/- 107.3 vs 161.6 +/- 97.7 mg/dl, p < 0.01) in the CHD male population between ages 25 and 69. In CHD females, triglycerides were higher (116.9 +/- 56.2 vs 91.5 +/- 43.3 mg/dl, p < 0.05) between ages 25 and 49; cholesterol showed no difference at any of the ages studied. HDL-C was much lower in both sexes of CHD patients at all ages studied (p < 0.001). Uric acid was higher in CHD males between ages 25 and 49 (p < 0.05), this significance being lost in the older age CHD group. Other components such as glycated hemoglobin, glucose and ionized calcium, were not different from those of the control group. PMID- 8246726 TI - [A proposal for spirometry standardization in the city of Buenos Aires]. AB - This paper refers to a Multicentric Quality Control Program on spirometry performed in Buenos Aires city. Seventeen spirometers (9 water seal, dry rolling seal or bellows, 5 flow detection type and 3 peak-flow meters) owned by 8 different Hospitals were tested. Reference values were obtained by means of an Explosive Decompression Simulator calibrated against a Stead-Wells bell spirometer, which meets ATS recommendations. Only 42% (6/14) of the tested spirometers agreed with the ATS minimal requirements with regard to FVC and FEV1; volumetric spirometers performed better than flow detectors. The maximal error was 11% when measuring a FVC of 3.41 L. Peak flow showed a very high inter instrument coefficient of variation. The need of frequent quality control checks is stressed. PMID- 8246727 TI - [E rosette inhibition in alcoholic liver disease]. AB - Various studies performed in chronic alcoholic patients have demonstrated immunologic alterations, which have been described more often in patients with alcoholic hepatitis or liver cirrhosis. We have observed that the serum of some patients with alcoholic liver cirrhosis produces the inhibition of E rosette formation by T lymphocytes. This observation induced us to study E rosette formation and its probable inhibition by the serum of chronic alcoholic patients. Subjects were split into three groups: Group 1: n = 21. Normal individuals. Group 2: n = 15. Chronic alcoholic patients without cirrhosis. Group 3: n = 26. Chronic alcoholic patients with histologically demonstrated liver cirrhosis. E rosette and E rosette inhibition (TIRE) sera tests were performed utilizing subject's sera tested against lymphocytes of normal individuals not related to group 1. The results found are listed in detail in Table 1 (mean = -SD) and Figure 1 (median), for each of the test groups. We applied unifactorial variance analysis and observed highly significant differences between the groups studied in all tests performed. It was found that tests that utilized I.E. discriminate most efficiently among the three groups of patients and that those which utilize unabsorbed assay serum (S/A) yield the best differentiation. Using this last assay it was observed that 20/21 control individuals showed less than 15% inhibition. On this basis, we decided to separate the results into 15% regular intervals (Table 3). Inhibition above 30% was found only in cirrhotic patients with the exception of one individual of the control group and one non-cirrhotic alcoholic patient with no alcoholic liver disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246728 TI - [Antipyretic effect of indomethacin vs aspirin in fever of tumor origin]. AB - Fever is frequently a symptom in patients suffering from cancer and in most cases it is related to infections or complications of the treatment. Some cancers can also be the direct cause of fever. A total of 28 episodes of fever in 8 patients with cancer were studied. The diagnoses were: 3 patients with lung cancer, 1 patient with chronic myelogenous leukemia, 1 patient with kidney cancer, 2 patients with non-Hodgkin lymphoma, and 1 patient with Hodgkin's disease. Were included cancer diagnosed patients of any age and sex, with three or more episodes of fever of more than 37.5 C (with a case of 38.5 C or more) after having eliminated any infectious etiology or fever caused by drugs. Were not excluded any patients who had received whole blood or blood derivative transfusions, chemotherapy or antibiotic treatment up to 48 hs before the fever peak. The patients were given intramuscularly 500 mg of aspirin or 50 mg of indomethacin. The first response in the temperature curve was evaluated while checking the axillary temperature six hours after drug administration and 48 hs later if the fever persisted. The patients who were given aspirin at first, were then given indomethacin and vice versa, using the same criteria to evaluate the response. In patients treated with indomethacin the temperature diminished quickly and completely (Fig. 1), unlike the effect achieved with the use of aspirin (Fig. 2). All patients treated with indomethacin also showed a remarkable clinical improvement which was not observed when aspirin was used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246729 TI - Evaluation of an attenuated Trypanosoma cruzi strain in rats. Analysis of survival, parasitemia and tissue damage. AB - Infection and tissue damage induced by parasites of an attenuated Trypanosoma cruzi culture strain (TCC) were studied in "I" line of inbred rats. Suckling rats (S), 3-5 day old were inoculated i.p. with 10(6) TCC (S1), 10(7) TCC (S2) and 10(8) TCC (S3). Weaned rats (W), 21-25 day old were inoculated s.c. with 10(6) TCC (W1), 10(7) TCC (W2) and 10(8) TCC (W3). The cultures yielded up to 2% of trypomastigotes. Controls inoculated either i.p. or s.c. with 10(6) blood form trypomastigotes (SC and WC) as well as normal controls (NC) were included. Survival was 100% in S1, S2 and S3, and 0% in SC on day 13 post-infection (p.i.). The latter animals died with acute Chagas disease signs. Survival was 100% in the W groups. In the first 30 days p.i. parasites were detected in S1, S2 and S3 and W1, W2, W3 groups after exhaustive examination. Parasites were easily found in WC and SC until day 13. Xeno-diagnoses were positive (5/5) at 2 months p.i. and negative at 6 months p.i. (W1, W2, W3, 0/23; WC, 0/5). Only cardiac lesions were slightly increased. The frequency of focal chronic myocarditis seemed to be increased in a dose-independent manner (S1, S2, S3, 26%; W1, W2, W3, 46%) but was not significant in comparison with NC, and even was lower than usually found in WC (61.3%). The reduced virulence and pathogenicity suggest that the TCC strain suffered a remarkable attenuation after long term in vitro culture. PMID- 8246730 TI - [Free erythrocyte protoporphyrin as a function of age in growing rats]. AB - The assessment of the iron (Fe) status is very important because its deficiency is one of the most common in both developing and industrialized countries, being particularly prevalent among infants and young children. Diagnosis is difficult in the presence of other conditions which interfere with the interpretation of laboratory tests, such as hemoglobin (Hb), hematocrit (Hct), serum iron, transferrin saturation percentage and serum ferritin. Free erythrocyte protoporphyrin (FEP) is a precursor of Heme and normally occurs in very low concentration in red blood cells (RBC); elevated values indicate early impaired iron nutritional status, providing information about gradual changes in the iron supply to the marrow. This laboratory test is a practical and convenient method because it needs a small blood sample easily preserved. Although the Second National Health and Nutritional Examination Survey (NHANES II) of USA has provided a good opportunity to define more precisely the cut-off points, it is doubtful whether the age-related differences in children represent normal development or the effects of iron deficiency. In order to provide information about these aspects the present experimental model was performed: weanling Wistar rats were fed until 95 days of age (t 95) with an isocaloric diet containing 20 or 30 g/100 g. of protein (casein) (N 20 and N 30, respectively), allowing the highest growth and erythropoiesis rates and covering the requirements of all the nutrients (Table 1). Body weight and food intake were recorded three times/week.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246731 TI - [Dilated cardiomyopathy and hypereosinophilia in a young female patient]. AB - A 23-year-old white woman was admitted because of cardiac failure (functional class II), palpable purpura and hypereosinophilia. A month before, she had been operated due to right femoral embolus. In this occasion, enhanced cardiac size on X-ray film of the chest and 12,300 eosinophils/mm3 were observed. She was asthmatic from her childhood. The laboratory data did not detect an etiology of hypereosinophilia. Serologic test for collagenous diseases, Chagas, hydatidosis, toxocariasis, Coxsackie B 1-6 and cytomegalovirus were not considered reactive. Three parasitologic stool examinations were negative. Duodenal sounding was negative. Myocardial dilatation was confirmed by mode M and B echocardiogram (60 mm left ventricular diastolic diameter), and by 99Tc radiocardiogram (left ventricular ejection fraction -LVEF-: 29% and right VEF: 15%). On the 25th day of treatment with 16-beta-methyl-prednisone (1 mg/K/d), eosinophil count was reduced to 300/mm3. On the 45th day, clinical improvement, dilatation reduction on echocardiogram and a 32% LVEF were observed. On the 10th month, the patient has no signs or symptoms of cardiac failure under treatment. Although endomyocardial biopsy was not performed but considering the low prevalence of myocardial dilatation at such an age, it is possible to postulate that the patient has undergone the initial necrotic stage (dilated cardiomyopathy) and the intermedial thrombotic stage (femoral embolus) of the eosinophilic endomyocardial disease. PMID- 8246732 TI - [Systemic lupus erythematosus, thrombocytopenia, and hemorrhages]. PMID- 8246733 TI - [Diet as a risk factor for coronary atherosclerosis in a rural area of Argentinian Patagonia. Comparison with the urban zone of Viedma]. AB - To determine risk factors, a clinical, cardiologic, biochemical and nutritional study was carried out. A sample of the rural population of Comi-Co (N = 92), which included descendants of Mapuche indians, was compared with a sample of the urban population of Viedma (N = 94). Sheep, goat and horse meat from the rural district was analyzed. In horse meat, intramuscular fat was 1.16 g%, with 34.5% of polyunsaturated fatty acids including linolenic (18:3 w3) 11.9%, linoleic (18:2 w6) 21.0%. The diet intake of the rural population was very different from that of the urban population (p < 0.001). It contained 3420 +/- 1027 vs 2110 +/- 654 kcal, carbohydrates 40 +/- 8 vs 55 +/- 10%, lipids 33 +/- 5 vs 24 +/- 8%, proteins 28 +/- 6 vs 20 +/- 6%, cholesterol 1021 +/- 380 vs 385 +/- 156 mg/day. The rural population showed a tendency to suffer cholelithiasis. Comi-Co vs Videma males had a CHDL 53 +/- 13 vs 47 +/- 9 mg/dl (p < 0.05) and no significant differences were found between both groups when it came to blood glucose, uric acid, triglycerides and Apo B. The frequency of individuals with CT > or = 240 mg/dl, CLDL > or = 160 mg/dl, cigarettes > or = 10/day seems to be less in the rural district. In Viedma vs Comi-Co males, with any one primary factor there was a ratio of 45% vs 24% (p < 0.05). The proportion of males with two risk factors was low. No individuals with three risk factors were found.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246734 TI - [Osteoporosis. Detection, prevention, and treatment]. AB - The osteoporotic syndrome is characterized by a reduction in quantity (osteopenia) and damage of the quality (microarchitecture) of bone mineralization. These are heterogeneous disturbances caused by multiple not well known metabolic causes. It is diagnosed by the presence of bone deformations or non-traumatic fractures. The early detection of osteopenia is important and it is considered that individuals whose bone mineral density (BMD) is lower than 80% of the normal value, are more exposed to fractures. Serum alkaline phosphatase, hydroxyproline and urinary pyridolines are biochemical markers. Nutritional aspects and physical movements are emphasized to obtain a larger bone mass, specially during the growing period. It is absolutely necessary to ingest enough quantity of calcium and in the southern regions of the country consumption of Vitamin D must be increased. Therapeutical resources tend to prevent osteopenia more effectively. Estrogens are greatly used in postmenopausic women for the prevention but not for the treatment of osteoporosis once established. They are effective but there still exist some doubts about the innocuousness of long term treatments. Calcitonines increase vertebral mineralization temporarily and may be used as alternatives of replacement hormonal therapies. Bisphosponates present different mechanisms, for example, pamidronate is not cytotoxic and for this reason does not interfere in the remodelling process, which is absolutely necessary to maintain or improve the architectural quality of the bone. The effect of bisphosphonates on mineralization is lasting. As they are new drugs, they have not yet been definitely approved in every country. Fluorine salts have always produced controversial results, some favourable initial effects do not prevent adverse consequences in a long treatment. Vitamin D, gallium nitrate and ipriflavone produce fairly good results, which must still be compared with those of the more active agents. The use of other drugs (tamoxifeno, parathormone, growth factors, thiazide and inhibitors of the proton pump) is still under development. It is concluded that osteoporosis is a pathologic process of ageing, for which there are pharmaceutical products available, specially to prevent osteopenia and bone damage. PMID- 8246735 TI - [Adjustment of the basal level of blood pressure]. AB - Basically the circulation must satisfy three requirements: 1) to provide an adequate blood flow to the tissues for maintaining their energetic needs, 2) to sustain renal function (glomerular pressure and renal blood flow) for the precise homeostasis of body fluids, and 3) to grant cutaneous circulation for controlling body temperature. Therefore, the arterial circulation can be separated in oxygen dependent, filtration dependent and thermic dependent sectors. The blood flow distribution through these regions depends on the myogenic tone of the resistance vessels. The oxygen dependent section receives 70% of the cardiac output and settles the equivalent resistance of the arterial tree; so, it is the main one responsible for setting the blood pressure level. The total resistance of this section should be adjusted to maintain the ATP/ADP relationship. The mechanisms involved in the regulation of the myogenic tone are local metabolic products (pO2, pCO2, pH, etc.), vasoactive substances present in the vascular wall (EDRF, AgII, PGs, etc.) and intracellular variations (Na++, Ca++, PKC, IP3, etc.). The vascular resistance of this section, adjusted as an electronic module, settles the minimum blood pressure needed to maintain the energetic equilibrium, independently of the pressure required to achieve a normal renal function. Thus, the kidney to fulfill its function must modulate this previously established myogenic tone by employing the renin angiotensin system. Circulating AgII will increase the vascular tone in the oxygen dependent section overriding its local controls until the blood pressure reaches the necessary level to maintain an adequate renal function. PMID- 8246736 TI - [1992 Nobel Prize of Medicine and Physiology: Edwin Krebs and Edmond Fisher. Protein phosphorylation and a hot summer]. PMID- 8246737 TI - [Science and medical education]. PMID- 8246738 TI - [Medical system--patient; patient-physician retroaction]. PMID- 8246739 TI - [Temporal arteritis, glossopharyngeal neuralgia, and syncope]. PMID- 8246740 TI - [Precordial pain and chronic Chagas disease]. PMID- 8246741 TI - [In memoriam of Leonidas de Mello Deane]. PMID- 8246742 TI - [Leonidas Deane]. PMID- 8246743 TI - Vitellogenesis in insects and other groups--a review. AB - The eggs from oviparous organisms contain large amounts of vitellus, or yolk, which are utilized by the growing embryo. Vitellogenesis is the process of vitellus accumulation and involves massive heterosynthetic synthesis of the protein vitellogenin (Vg) and its deposition in the oocyte. This work summarizes data on Vg structure, synthesis, uptake by oocytes and its fate during embryogenesis. The hormonal control of vitellogenesis and its tissue, sex and temporal regulation are also discussed. Where it is available, data on structure and expression of Vg-coding genes are reviewed. Insect vitellogenesis is priorized although other oviparous animal groups outside insects are also treated. PMID- 8246744 TI - Epidemiological aspects of American cutaneous leishmaniasis in a periurban area of the metropolitan region of Belo Horizonte, Minas Gerais, Brazil. AB - In order to characterize the epidemiology of American Cutaneous Leishmaniasis (ACL) in a periurban area of the municipality of Sabara in the metropolitan region of Belo Horizonte (MRBH), an area until then considered free of the disease, a cross sectional survey was undertaken in 1990. The survey of the population consisted of 1119 interviews and 881 clinical examinations using Montenegro's skin test (MST). A low prevalence (3.7%) of positive MST was encountered. The disease had been occurring in the area for about 20 years in the form of sporadic cases. The predominant species of sandfly both in domestic areas and nearby areas of secondary vegetation was Lutzomyia whitmani. A canine survey of delayed hypersensitivity to the antigen P10,000 identified only one dog with a positive reaction out of 113 examined. The transmission of ACL in MRBH was confirmed. The occurrence of the disease in women, children and individuals with no contact with forest areas as well as the presence of potential vector species in the domiciliar environment, suggests the transmission of the disease in this environment. PMID- 8246745 TI - Histopathological alterations induced by non-viable cells and biochemical fractions from Paracoccidioides brasiliensis in mice. AB - Non-viable cells and biochemical fractions from Paracoccidioides brasiliensis were obtained for experimental inoculation in mice and posterior histopathological analysis. Dead total fungus, total fungus disrupted by sonorous waves, lipids of the fungus, supernatant of the lipid purification, integral and disrupted fungus free of lipids were obtained. The six preparations arose from masses of lyophilized yeasts of a recent isolate of P. brasiliensis (strain JT-1) and from a "Pool" equitably constituted by four strains maintained in laboratory for a long time (SN, 2, 18 and 192). Different doses of the 12 preparations were intraperitoneally inoculated and histopathological analysis were done 30 days later. This analysis showed that all the inoculated preparations gave origin to inflammatory foci, except the one designated "supernatant of lipid purification". The alterations were detected exclusively in the liver of the animals and occurred from the smallest dose tested (1 mg), with exception of the lipids of the fungus, where the foci appeared only from a 3 mg dose onwards. No difference in the capacity of inducing histopathological alterations was found between the preparations obtained from the recent isolate (JT-1) and from the older ones ("Pool"). On the other hand, an increase of the number of inflammatory foci in function of the inoculated dose was observed. PMID- 8246746 TI - Standardization of the dot enzyme-linked immunosorbent assay (dot-ELISA) for experimental plague. AB - A dot enzyme linked immunosorbent assay (dot-ELISA) was previously developed to detect specific antibodies in rabbits sera immunized against F1A protein obtained from Yersinia pestis. This antigen was covalently linked onto the surface of dacron (polyethyleneterephthalate). Here, standard conditions are described for the optimization of this procedure: an amount of 20 ng of F1A protein was fixed onto dacron; anti-rabbit IgG peroxidase conjugate diluted 1:8,000 and 30% non-fat instant milk as blocking substance were used throughout the method. This procedure was compared with that employing nitrocellulose as solid-phase which showed to be more sensitive. However, the method based on dacron did not show false positive reactions against non-immunized rabbits sera at low antigen amount and diluted anti-IgG peroxidase conjugate. PMID- 8246747 TI - Partial isolation and some properties of enterotoxin produced by Bacillus cereus strains. AB - Extracellular proteins produced by Bacillus cereus AL-42 and AL-15 were fractioned by chromatography on QAE-Sephadex and Sephadex G75. This last chromatographic process resulted in three peaks. The major peak showed vascular permeability activity to rabbits, lethality to mice, and cytotoxicity to Vero and Hela cells. The analysis by SDS-PAGE after ultrafiltration confirm recent findings that the enterotoxin is a compound with molecular mass > 30,000. PMID- 8246748 TI - Use of a selective medium with potassium tellurite to follow intestinal colonization of hospitalized patients by drug-resistant Enterobacteriaceae. AB - Nosocomial infections are a relevant factor in complicating the recovery of patients interned for even minor causes. In a attempt to determine their origin it is crucial to consider that their origin is of an endogenous nature. Looking for an accessible expression of intestinal colonization we analyzed fecal samples from 3 separate groups of hospital patients collected after different lengths of time. For practical reasons one group was studied prospectively and two other groups (patients hospitalized for up to 7 days and patients hospitalized for more than 7 days) were compared to one another. We looked for the emergence of tellurite resistance among Enterobacteriaceae using a selective medium, MacConkey potassium tellurite (MCPT). The frequency of prospectively studied patients with tellurite resistant strains was significantly greater after 7 days of hospitalization. For the two other groups, patients with more than 7 days of hospitalization showed a significant increase of bacterial species and of strains with new antimicrobial resistance markers. High molecular weight plasmids were detected in some of these strains. These data show that the MCPT medium is a useful tool for the investigation of bowel colonization in hospitalized patients by drug-resistant Enterobacteriaceae. PMID- 8246749 TI - Morphological and virological studies in six autopsies of children with adenovirus pneumonia. AB - Pulmonary lesions compatible with adenovirus infection were detected by gross and microscopic examination of autopsy tissues from children aged from 5 to 34 months. Hepatic lesions indicative of systemic infection were also found in four of the children. The viral etiology was confirmed in three cases by in-situ hybridization, electron-microscopy and immunofluorescence performed in paraffin embedded tissues, and in one case by cell culture isolation of adenovirus type 2 from nasopharyngeal exudate. Routine testing by methods additional to conventional light microscopy would probably have revealed a larger number of adenovirus infections among the 1,103 autopsy records analyzed in this study. PMID- 8246750 TI - An experimental and clinical assay with ketoconazole in the treatment of Chagas disease. AB - Ketoconazole, an azole antifungic drug which is already in the market has also been demonstrated to be active against Trypanosoma cruzi experimental infections. In this paper we confirmed the drug effect and investigated its range of activity against different T. cruzi strains naturally resistant or susceptible to both standard drugs Nifurtimox and Benznidazole used clinically in Chagas disease. Moreover, we have shown that the association of Ketoconazole plus Lovastatin (an inhibitor of sterol synthesis), which has an antiproliferative effect against T. cruzi in vitro, failed to enhance the suppressive effect of Ketoconazole displayed when administered alone to infected mice. Finally, administration in chronic chagasic patients of Ketoconazole at doses used in the treatment of deep mycosis also failed to induce cure as demonstrated by parasitological and serological tests. The strategy of identify and test drugs which are already in the market and fortuitously are active against T. cruzi has been discussed. PMID- 8246751 TI - Pharmacokinetic profile of two different pharmaceutical forms of theophylline (a slow release tablet and a syrup) after multiple dose administration to healthy human volunteers. AB - Due to the narrow therapeutic range of theophylline, plasma concentrations of this drug are monitored in patients undergoing chronic therapy. Slow-release preparations avoid the fluctuations in plasma levels and improve patient compliance. In this study, we have compared the pharmacokinetic profiles of a theophylline slow-release tablet and a syrup form, when administered in multiple doses to healthy adult volunteers. The classification based upon releasing patterns is confirmed. PMID- 8246752 TI - Reversion of culture-induced virulence-attenuation in Trypanosoma cruzi. PMID- 8246753 TI - Copy number differences in the 195 bp repeated satellite DNA from Trypanosoma cruzi and Trypanosoma rangeli: potential use for epidemiologic surveys. PMID- 8246754 TI - A simple protocol for the physical cleavage of Trypanosoma cruzi kinetoplast DNA present in blood samples and its use in polymerase chain reaction (PCR)-based diagnosis of chronic Chagas disease. PMID- 8246755 TI - Dispersive flight and house invasion by Triatoma guasayana and Triatoma sordida in Argentina. AB - Flight activity and invasion of houses by Triatoma sordida and T. guasayana were studied in the Province of Santiago del Estero, Argentina. Spontaneous findings of both species in houses were recorded from 1982 to 1989. Light trap collections were performed in 1982, 1983 and 1984, at the woods surrounding the settlements of Amama (43 houses) and Trinidad (19 houses). Most of the 101 triatomines collected, were unfed and negative for Trypanosoma cruzi. T. guasayana predominated over T. sordida, and both appeared on the lighted screens between 19 31 min (mean 24) after dusk and the catch time was 30-45 min. Although entomological evaluation of 41 houses at Amama performed in September 1985, just before insecticidal spraying, showed that Triatoma infestans predominated, adults of T. guasayana were collected in sleeping places, in 7 houses (17%). Most triatomines invading houses from then up to 1990 were flying T. guasayana (20/27) and females outnumbered males. Three non-infected T. guasayana females were fed on man and two T. guasayana males positive for "T. cruzi like" trypanosomes were unfed. Therefore, visiting hungry adults could transmit T. cruzi to people and introduce wild parasites to the domestic cycle. T. guasayana stands as the main potential substitute of T. infestans in the studied area, and it might play there the same role as T. sordida in Brazil. PMID- 8246756 TI - Cytological and isoenzyme analysis of the Bucay and Quevedo cytotypes of the Onchocerciasis vector Simulium exiguum (Diptera: Simuliidae) in Ecuador. AB - Four cytotypes of Simulium exiguum occur in Ecuador, where this morphospecies is the primary vector of onchocerciasis. In this paper, we give the first full description of the banding pattern of the larval polytene chromosomes of the Quevedo cytotype and assess its degree of genetic separation from the Bucay cytotype. Both cytotypes differ from the chromosomal standard sequence (of the Cayapa cytotype) by the fixed inversions IIL-5 and IIL-6. The Quevedo cytotype additionally differs from the standard and Bucay cytotypes by possessing a differentiated X chromosome, which is indicated by the inversion IIS-A. As the degree of reproductive isolation between the Bucay and Quevedo cytotypes has not yet been established, they must be regarded as intraspecific variants of the same species. In fact, isoenzyme characterizations showed that the Bucay and Quevedo cytotypes are differentiated only to the extent expected of incipient species or geographical populations. Moreover, the sibling species status previously given to the Bucay cytotype needs be reassessed, there being inadequate analysis from areas in Ecuador where Bucay occurs in sympatry with the standard Cayapa cytotype. No isoenzyme electromorphs were discovered that identified all or most adult females of any one (cytotypepure) collection. PMID- 8246757 TI - The distribution of intestinal helminth infections in a rural village in Guatemala. AB - Fecal egg count scores were used to investigate the distribution and abundance of intestinal helminths in the population of a rural village. Prevalences of the major helminths were 41% with Ascaris lumbricoides, 60% with Trichuris trichiura and 50% with Necator americanus. All three parasites showed a highly aggregated distribution among hosts. Age/prevalence and age/intensity profiles were typical for both A. lumbricoides and T. trichiura with the highest worm burdens in the 5 10 year old children. For hookworm both prevalence and intensity curves were convex in shape with maximum infection levels in the 30-40 year old age class. Infected females had higher burdens of T. trichiura than infected males in all age classes of the population; there were no other effects of host gender. Analysis of associations between parasites within hosts revealed strong correlations between A. lumbricoides and T. trichiura. Individuals with heavy infections of A. lumbricoides and T. trichiura showed highly significant aggregation within households. Associations between a variety of household features and heavy infections with A. lumbricoides and T. trichiura are described. PMID- 8246758 TI - Morphological characterization of the hemocytes of the pulmonate snail Biomphalaria tenagophila. AB - The blood cells of the pulmonate snail Biomphalaria tenagophila, an important transmitter of the trematode Schistosoma mansoni in Brazil, were examined by light and transmission electron microscopy (TEM). Two hemocyte types were identified: hyalinocytes and granulocytes. Hyalinocytes are small young (immature), poorly spreading cells, which have a high nucleocytoplasmic ratio and are especially rich in free ribosomes. They do not appear to contain lysosome like bodies and represent less than 10% of the circulating hemocytes. Granulocytes are larger hemocytes which readily spread on glass surface and which strongly react to the Gomori substrate, indicating the enzyme acid phosphatase usually found in lysosomes. Ultra-structurally, they contain a well-developed rough endoplasmic reticulum, dictyosomes and some lysosome-like dense bodies. Granulocytes do not exhibit a characteristic granular aspect and the few granules observed in the cytoplasm should correspond to a lysosome system. They were named granulocytes instead of amoebocytes to use the same terminology adopted for Biomphalaria glabrata in order to make easier comparative studies. This is a preface study for more specific investigations on the functional activities of the blood cells of B. tenagophila and their interactions with the trematode parasite. PMID- 8246759 TI - Response of drug resistant isolates of Schistosoma mansoni to antischistosomal agents. AB - The susceptibility of four isolates of Schistosoma mansoni (BH, MAP, MPR-1 and K) to four multiple doses of anti-schistosomal agents (hycanthone, niridazole, oxamniquine, and praziquantel) were evaluated in infected female Swiss albino mice. These schistosomal isolates had been maintained in the laboratory without further drug pressure for 20 to 30 generations. Multiple dosage regimens were used for each drug against each isolate of S. mansoni to generate ED50 (effective dose 50%) values. Results demonstrated that the K isolate is resistant to niridazole, the MPR-1 isolate to oxamniquine, and the MAP isolate to both hycanthone and oxamniquine. The BH isolate was susceptible to all drugs and was used as the reference isolate. All isolates were susceptible to parziquantel. The significance of the difference in response of the MPR-1 and MAP isolates is discussed. These results confirm the resistance of these isolates of S. mansoni to three schistosomicides and demonstrate that the resistance of these isolates are stable over long periods of time without exposure to drugs. PMID- 8246760 TI - Evaluation of spleen cell population and effect of splenectomy on granuloma modulation in BALB/c mice infected with Schistosoma mansoni. AB - A kinetic study of the cells present in the spleen of BALB/c mice infected with Schistosoma mansoni was carried out. The lymphocytes were evaluated phenotypically with monoclonal antibodies and the effect of splenectomy on the modulation of periovular granuloma was also investigated. The infected mice had proportional increases in the numbers of neutrophils, plasma cells, macrophages and eosinophils in the spleen. The largest number of neutrophil, plasma cells and macrophage were found between the 8th and the 12th week of infection, while the amount of eosinophils were higher later on, around the 20th week. The lymphocytes phenotypically characterized as Thy 1.2, Lyt 1.2 (CD4) increased mildly in proportional numbers. However, the percentage of lymphocytes with the Lyt 2.2 (CD8) phenotype, which is characteristic of suppressor and cytotoxic T cells, increased significantly with the progress of the disease. The numbers of B lymphocytes, which comprise 50% of the mononuclear cells present in the spleen, increased significantly till the 16th week when they began to decrease. The mean diameters of periovular granulomas were comparatively similar in both experimental groups (splenectomized and non-splenectomized mice). However, the evolutive types of granuloma (exudative, intermediate and fibrous) in splenectomized mice were proportionally different from those of non splenectomized mice in the 16th and 24th week of infection. It is inferred that lymphnodes or other secondary lymphoid organs, in the absence of the spleen, assume a modulating action on periovular granulomas, although the evolution of the granulomas is somehow delayed in splenectomized mice. PMID- 8246761 TI - Alteration of tissue vanadium content in diabetes. AB - A great deal of interest in the element vanadium has been generated recently because of its potential as a therapeutic agent for diabetes mellitus. Vanadium's insulin-mimetic properties and its requirement for proper growth and development suggest that it may be involved in insulin's mechanism of action. We have therefore examined vanadium levels in kidney, muscle, and liver tissues from normal and diabetic BB Wistar rats. Our results indicate that diabetes mellitus can decrease the tissue vanadium content of liver, suggesting that the trace element vanadium may be important in insulin action. PMID- 8246762 TI - 3-Deazaadenosine and MDL29350 differentially affect the methylation of serine-and ethanolamine-derived phosphatidylethanolamine in Hep G2 cells. AB - The effect of 3-deazaadenosine (DZA) and the hypolipidemic drug MDL29350 (2-[3,5 di(t-butyl-4-hydroxyphenyl)thio]hexanoic acid) on the synthesis and methylation of phosphatidylethanolamine (PE) originating from the cytidine diphosphate (CDP) ethanolamine pathway and PE originating from decarboxylation of phosphatidylserine (PS) was investigated. DZA and MDL29350 did not affect the synthesis of PE by either pathway; however, methylation of ethanolamine-derived PE was inhibited by 80% and methylation of serine-derived PE was inhibited by 36% by 20 mumol/LDZA or MDL29350. The differential inhibition of the methylation of PE synthesized via serine or ethanolamine suggests that in Hep G2 cells PE-N methyltransferase (PENMT) may be segregated into distinct compartments that are differentially accessible to the drugs. PMID- 8246763 TI - Overfeeding-induced obesity in rats: insulin sensitivity and autonomic regulation of metabolism. AB - The metabolic consequences of the development of obesity and the underlying mechanisms were investigated. For this purpose, male rats were overfed for 5 weeks through long-term gastric catheters. Permanent cardiac cannulas implanted before the overfeeding period allowed frequent blood sampling and infusions without disturbing the rats. Hyperalimented rats became grossly obese, displayed elevated basal plasma norepinephrine (NE) concentrations, and developed hyperinsulinemia and insulin insensitivity, but remained normoglycemic and preserved normal intravenous (IV) glucose tolerance. During physical exercise (ie, 15 minutes of swimming), obese rats displayed exaggerated increases in blood glucose concentrations, whereas plasma free fatty acid (FFA) responses were blunted. These alterations were probably due to decreased NE release by the sympathetic nervous system during exercise and to altered tissue responsivity to adrenergic stimulation. The latter was demonstrated by infusions of catecholamines in the resting state. Responses to mild stress were increased in obese animals, as indicated by increased responses of plasma epinephrine (E) and corticosterone during handling and first contact with water. The results of the present study indicate that overfeeding induces changes in the sympathetic control of metabolism and insulin secretion. Whereas elevated NE levels in the basal state probably reflect increased energy expenditure, the pattern of nutrient mobilization during exercise is directed toward sparing of fats. PMID- 8246764 TI - Growth hormone treatment improves serum lipids and lipoproteins in adults with growth hormone deficiency. AB - The effects of 6 months' treatment with recombinant human growth hormone (rhGH) on serum lipids and lipoproteins were assessed in 24 adult patients with GH deficiency in a double-blind, placebo-controlled trial. Compared with age-, weight-, and sex-matched controls, the patients had significantly higher serum concentrations of total cholesterol (P = .002), low-density lipoprotein (LDL) cholesterol (P < .001), apolipoprotein B ([apoB] P = .011), and triglyceride (P = .017), and lower concentrations of high-density lipoprotein (HDL) cholesterol (P < .001). The prevalence of elevated serum cholesterol, triglyceride, LDL cholesterol, and apo B levels was 39%, 26%, 39%, and 25%, respectively; 75% of patients had decreased concentrations of HDL cholesterol. Treatment with rhGH (0.07 U/kg daily) resulted in decreases in total cholesterol level (5.8 +/- 0.3 to 5.1 +/- 0.3 mmol.L-1 over 6 months; P = .01 compared with placebo), LDL cholesterol level (4.22 +/- 0.25 to 3.19 +/- 0.23 mmol.L-1; P = .0003), LDL:HDL cholesterol ratio (5.57 +/- 0.47 to 3.29 +/- 0.33; P = .03), apo B level (1.07 +/ 0.06 to 0.84 +/- 0.07 g.L-1; P = .003), and apo B: A-1 ratio (0.73 +/- 0.05 to 0.69 +/- 0.05; P = .01). HDL cholesterol and apo A-1 levels did not change following rhGH treatment. The changes in lipid and lipoprotein levels were not significantly related to changes in insulin, thyroid hormones, insulin-like growth factor-1 (IGF-1), or indices of adiposity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246765 TI - Unidirectional transfer in vivo of high-density lipoprotein cholesteryl esters to lower-density lipoproteins in the pig, an animal species without plasma cholesteryl ester transfer activity. AB - The metabolism of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesteryl esters (CE) was studied in the pig, an animal species without plasma cholesteryl ester transfer activity (CETA). In the first series of experiments, LDL and HDL from normocholesterolemic pigs were radiolabeled with cholesteryl (1-14C)oleate and intravenously administered to two groups of four normocholesterolemic pigs. Radioactive tracer in LDL remained associated with the LDL fraction, and there was no transfer of LDL-CE to HDL. The transport rate (which represents the production and disposal rate) of LDL-CE in normocholesterolemic pigs was 39 mumol CE/h/L. However, radiolabeled HDL-CE were transferred to LDL (25%), and 36% of the LDL-CE mass was derived from the HDL. The transport rate of HDL-CE was 54 mumol CE/h/L, and the flux of HDL-CE to LDL was 14 mumol CE/h/L. There was no accumulation of radiolabeled HDL-CE in very-low density lipoprotein (VLDL), which suggests that there was no transfer to VLDL. However, this does not rule out the possibility that either the very low levels of VLDL-CE (< 0.09 mmol/L) or the rapid turnover rate of the VLDL pool might have prevented the accumulation of substantial amounts of tracer in VLDL. Therefore, in a second set of experiments, the kinetics of HDL-CE were studied in high-fat and high-cholesterol-fed pigs with elevated VLDL-CE concentrations (1.92 mmol/L). Hypercholesterolemia was associated with increased transport rates of LDL-CE (165 mumol/h/L) and HDL-CE (78 mumol/h/L) and with an increased flux of HDL-CE to LDL (78 mumol/h/L).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246766 TI - Modeling plasma lipoprotein-bile lipid relationships: differential impact of psyllium and cholestyramine in hamsters fed a lithogenic diet. AB - Hamsters fed a lithogenic diet become hyperlipemic with elevated very-low-density lipoprotein (VLDL) and high-density lipoprotein 2 (HDL2) cholesterol pools and develop lithogenic bile in which chenodeoxycholate (cheno) typically predominates. The relationship between these distorted lipoprotein and bile lipid profiles and gallstone induction was investigated in male Syrian hamsters fed for 5 weeks a gallstone-inducing purified diet (5% butter, 0.4% cholesterol) or the same diet supplemented with 5% psyllium or 1% cholestyramine, agents known to alter bile acid metabolism. The gallstone diet essentially doubled plasma cholesterol level, whereas psyllium decreased it to near normal, and cholestyramine to a subnormal level, while correcting the distorted distribution of cholesterol among lipoproteins. Both the gallstone diet and psyllium produced cholesterol-laden livers, in contrast to subnormal values produced by cholestyramine. Fecal bile acid excretion was increased eightfold with cholestyramine and fourfold with psyllium relative to the value produced by the gallstone diet and a literature control value. Supersaturated bile developed with the gallstone diet (lithogenic index [LI], 2.3 +/- 0.6), whereas the LI was decreased by psyllium (1.2 +/- 0.4) and cholestyramine (0.7 +/- 0.3). The gallstone diet decreased the concentration of bile acids in gallbladder bile, but greatly increased the percentage of taurochenodeoxycholic acid, whereas psyllium preferentially decreased all taurine-conjugated bile acid levels and expanded glycocholate output. Cholestyramine greatly decreased the secretion of biliary cholesterol and cheno independent of its conjugation. Accordingly, psyllium increased the glycine to taurine ratio of gallbladder bile fivefold, whereas cholestyramine did not affect this ratio, but increased the cholate to cheno ratio dramatically (25-fold) as compared with a threefold increase with psyllium. This combination of biliary lipid and bile acid alterations induced coordinated responses in the LI and the hydrophobicity index (HI) such that cholesterol gallstones developed in 11 of 12 hamsters fed the gallstone diet, whereas only one of 11 of the psyllium-fed and none of 12 cholestyramine-fed hamsters had cholesterol stones. Thus, psyllium and cholestyramine differentially increased bile acid excretion, which improved the lipoprotein profile and inhibited cholesterol gallstone formation. Both agents operated by different means to decrease biliary cholesterol secretion and the percentage of cheno, which decreased the LI and HI, respectively. PMID- 8246767 TI - Insulin resistance is related to albuminuria in patients with type II (non insulin-dependent) diabetes mellitus. AB - To determine whether albuminuria is associated with insulin resistance in patients with type II (non-insulin-dependent) diabetes mellitus, we performed hyperinsulinemic (40 mU/m2/min) euglycemic clamp studies in patients with a urinary albumin excretion (UAE) rate greater than 30 mg/24 h and in patients with a UAE less than 30 mg/24 h. The UAE-positive group (n = 22) did not differ significantly from the UAE-negative group (n = 18) with respect to age, sex, treatment of diabetes, body mass index, fasting or postload blood glucose or plasma insulin levels, blood pressure, or known duration of diabetes. The mean glucose disposal rate (GDR) was significantly lower in the UAE-positive group than in the UAE-negative group (3.44 +/- 0.29 v 4.75 +/- 0.52 mg/kg/min, P < .05). When patients with hypertension were excluded, GDR was still markedly lower in the UAE-positive group than in the UAE-negative group (3.89 +/- 0.54 v 6.68 +/ 0.71 mg/kg/min, P = .01). The difference between groups persisted even after adjustment for body mass index, sex, and hypertension (ANCOVA; P < .05). These results indicate that the presence of microalbuminuria is associated with impaired insulin action in patients with type II diabetes mellitus. PMID- 8246768 TI - Insulin and glucose suppress hepatic glycogenolysis by distinct enzymatic mechanisms. AB - Both insulin and hyperglycemia can effectively suppress hepatic glucose output (HGO). We examined whether insulin and hyperglycemia specifically suppress liver net glycogen breakdown in a rat model in which glycogen is the major source of HGO. We further examined whether insulin and hyperglycemia act by similar or distinct enzymatic mechanisms. HGO, the rate of net glycogen loss, and glycogen phosphorylase and synthase activities were measured in fed, anesthetized rats infused with saline or insulin (7 mU/min/kg) while either maintaining plasma glucose at basal (7.8 +/- 0.2 mmol/L, euglycemic clamp [EC]) or at 10 mmol/L above basal (18 +/- 0.4 mmol/L, hyperglycemic clamp [HC]). During the basal period, the rate of HGO in each group was comparable to the rate of net glycogen breakdown, averaging 76 +/- 9 and 75 +/- 5 mumol/min/kg, respectively. Thus glycogen breakdown appeared to be a major source of ongoing HGO. Over the last 60 minutes of the experimental period, the rate of glycogenolysis averaged 69 +/- 8 mumol/min/kg in saline-treated rats; this could account for about 80% of the total HGO. During both EC and HC studies, HGO was suppressed (5.5 +/- 3 and -3.6 +/- 10 mumol/min/kg, respectively; P < .001 for each). Net glycogen breakdown decreased by 50% in EC rats (P < .05) and ceased in HC rats (P < .001). Glycogen synthase was predominantly in the active form in all three experimental groups (87% +/- 2%, 89% +/- 2%, and 95% +/- 3% in saline, EC, and HC rats, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246769 TI - Cholesterol and fatty acid metabolism in piglets fed sow milk or infant formula with or without addition of cholesterol. AB - Several studies have reported that plasma cholesterol and phospholipid (PL) levels of arachidonic acid (20:4n-6) are lower and PL levels of linoleic acid (18:2n-6) are higher in infants fed formula than in infants fed human milk. Plasma cholesterol level and possibly the dietary intake of cholesterol could be related to plasma PLn-6 fatty acid metabolism because plasma PL 18:2n-6 is used for esterification of plasma free cholesterol. Whether the low cholesterol content of infant formula as compared with human milk is related to the difference in plasma n-6 fatty acid levels between infants fed human milk and infants fed formula is not known. This study determined the effect of feeding formula with 0.05 mmol cholesterol/L, formula with 1.09 mmol cholesterol/L, or sow milk with 0.34 mmol cholesterol/L on plasma, liver, and bile lipid fatty acid levels and liver low-density lipoprotein (LDL) receptor mass in piglets. Liver microsomal 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase activity and plasma lathosterol were assayed as indices of liver and body cholesterol synthesis, respectively. Formula with or without cholesterol added, or sow milk, was fed from birth to 18 days of age. Providing cholesterol in the formula did not correct the significantly lower plasma cholesterol or plasma and liver PL 20:4n-6 levels associated with formula feeding. The liver total cholesterol and cholesteryl esters (CE), biliary bile acid, and PL concentrations were significantly higher and the liver HMG CoA reductase activity and plasma lathosterol:cholesterol ratio were significantly lower in piglets fed the formula with cholesterol than in piglets fed the formula without cholesterol added.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246770 TI - Effects of glucose, galactose, and lactose ingestion on the plasma glucose and insulin response in persons with non-insulin-dependent diabetes mellitus. AB - Galactose usually is ingested as lactose, which is composed of equimolar amounts of glucose and galactose. The contribution of galactose to the increase in glucose and insulin levels following ingestion of equimolar amounts of galactose and glucose, or lactose, has not been reported in people with non-insulin dependent diabetes mellitus (NIDDM). Therefore, we studied the effects of galactose ingestion alone, as well as with glucose either independently or in the form of lactose, in subjects with untreated NIDDM. Eight male subjects with untreated NIDDM ingested 25 g glucose, 25 g galactose with or without 25 g glucose, or 50 g lactose as a breakfast meal in random sequence. They also received 50 g glucose on two occasions as a reference. Water only was given as a control meal. Plasma galactose, glucose, glucagon, alpha-amino nitrogen (AAN), nonesterified fatty acids (NEFA), and serum insulin and C-peptide concentrations were determined over a 5-hour period. The integrated area responses were quantified over the 5-hour period using the water control as a baseline. Following ingestion of 25 g galactose, the maximal increase in plasma galactose concentration was 1 mmol/L. The mean maximal increases in plasma galactose concentration following ingestion of 25 g galactose + 25 g glucose or following 50-g lactose meals were similar and were only 12% of that following ingestion of galactose alone (P < .05). The mean galactose area response over the water control for the 25-g galactose meal was 0.95 +/- 0.31 mmol.h/L.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246771 TI - Gender influences counterregulatory hormone responses to hypoglycemia. AB - It has been generally assumed that counterregulatory hormone responses to hypoglycemia are not influenced by gender. To test this assumption, we analyzed three separate hypoglycemic insulin clamp studies in age-matched, healthy, non obese females (n = 33) and males (n = 37). In one study (12 females, 17 males), plasma glucose level was rapidly decreased to about 57 mg/dL for 100 minutes with a 0.65-mU/kg/min insulin infusion. Despite an identical decrease in glucose level, the increase in epinephrine (361 +/- 64 v 188 +/- 38 pg/mL, P < .05), norepinephrine (132 +/- 28 v 47 +/- 19 pg/mL, P < .01), and growth hormone ([GH] 16.0 +/- 3.8 v 4.9 +/- 1.9 ng/mL, P < .05) levels, but not glucagon or cortisol levels, were significantly greater in males than in females, respectively. In the second study (10 females, eight males), a 5.0-mU/kg/min insulin infusion was used to decrease glucose levels to 55 mg/dL for 180 minutes. Epinephrine (P < .05) and GH (P < .01) responses were greater in males than in females. In a third study (11 females, 12 males), plasma glucose level was gradually decreased to about 50 mg/dL over 240 minutes. Again epinephrine (P < .01), norepinephrine (P < .01), GH (P < .05), and cortisol (P < .01) responses were nearly twofold greater in males (P < .01). Multivariate analysis of all 70 subjects identified gender as the most significant factor contributing to the epinephrine (P < .001) and norepinephrine (P < .005) responses, and also as a significant contributor to the GH response (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246772 TI - Reciprocal changes in gluconeogenesis and ureagenesis induced by fatty acid oxidation. AB - Fatty acids produced a stimulation of gluconeogenesis and either inhibition or no effect on ureagenesis in livers perfused with gluconeogenic substrates and having NH4Cl plus ornithine as the nitrogen source. This finding indicates that stimulation of flux through pyruvate carboxylase is not sufficient to enhance urea production from ammonia. The metabolic action of fatty acids showed the following characteristics: (1) it was concentration-dependent, showing saturation type kinetics similar to those described for fatty acid oxidation; (2) the stimulatory action on gluconeogenesis was constant and independent of NH4Cl concentration, whereas the inhibition of ureagenesis was variable and dependent on NH4Cl concentration and the degree of reduction of the gluconeogenic substrate; and (3) fatty acids produced apparent reciprocal changes in the state of reduction of the cytosolic and mitochondrial NAD systems. Fatty acid oxidation exerted its effect mainly, if not exclusively, by preventing the gluconeogenic substrate-induced stimulation of ureagenesis. Fatty acids also inhibited ureagenesis without stimulating gluconeogenesis (lactate < 1 mmol/L), ruling out a limiting energy availability as the cause of the inhibition. One or both of the following two mechanisms seem to account for the fatty acid-induced inhibition of ureagenesis from NH4Cl. First, a decreased uptake of ornithine, and second, decreased flux through pyruvate dehydrogenase and probably other NAD(P)-linked mitochondrial dehydrogenases. The correlation found between the ability of fatty acids to inhibit ureagenesis and the state of activation of pyruvate dehydrogenase supports the latter point. PMID- 8246773 TI - Actions of the novel antidiabetic agent englitazone in rat hepatocytes. AB - We examined effects of a novel antidiabetic agent, racemic englitazone (CP 68,722, Pfizer), on normal rat hepatocytes in vitro. For optimal effects, CP 68,722 must be preincubated for approximately 20 minutes. CP 68,722 inhibited the actions of glucagon on glycogenolysis (measured by monitoring cyclic adenosine monophosphate [cAMP] levels, phosphorylase activation, and glucose output) and gluconeogenesis (from 14C-lactate). Since CP 68,722 was able to attenuate the ability of glucagon to increase cAMP levels, this may account for part of its inhibitory actions on glycogenolysis and gluconeogenesis. The observation that CP 68,722 also inhibits the ability of the cAMP analog, 8-(4-chlorophenylthio) adenosine 3':5'-cyclic monophosphate (8 CPT cAMP), to stimulate phosphorylase a is consistent with an effect of CP 68,722 to activate cAMP-dependent phosphodiesterase. The ability of vasopressin (an agonist known to stimulate glycogenolysis via a Ca(2+)-dependent mechanism) to stimulate phosphorylase a was slightly inhibited by CP 68,722. Another site of action of CP 68,722 was to inhibit hormonal-mediated Ca2+ influx, an effect that would decrease intracellular free calcium ([Ca2+]i), thereby inhibiting the actions of the Ca(2+)-dependent hormones such as alpha 1-adrenergic agonists and vasopressin, agents known to promote glucose output from the liver. In summary, CP 68,722 inhibits glucagon-stimulated glycogenolysis and gluconeogenesis in hepatocytes by a mechanism that may include activation of cAMP phosphodiesterase and inhibition of Ca2+ influx. PMID- 8246774 TI - Impact of regional and total body composition and hormones on resting energy expenditure in overweight postmenopausal women. AB - The independent impact of regional and total body composition and sex and thyroid hormone levels on resting energy expenditure (REE) was assessed in 121 healthy, overweight (body mass index [BMI], 25 to 42 kg/m2), postmenopausal women (aged 49 to 58 years). REE was measured by indirect calorimetry with a ventilated hood, and the total, trunk, and peripheral body composition by dual-energy x-ray absorptiometry (DXA). Sex and thyroid hormone levels were also measured. REE correlated significantly (P < .05) better with trunk lean tissue mass ([LTM] r = .61) than with total-body (r = .53) or peripheral LTM (r = .30). Trunk LTM (R2 = .37), total-body fat tissue mass ([FTM] R2 = .04), androstenedione (R2 = .03), and total triiodothyronine ([T3] R2 = .02) were all significantly independently associated with REE. Together they explained 46% (model R2) of the interindividual variation in REE, with a standard error of estimate (SEE) of 549 kJ/d. We conclude that the interindividual variation in REE is explained mainly by differences in the visceral component of LTM, with additional minor information from the total-body FTM, androstenedione, and T3. PMID- 8246775 TI - Unusual high-density lipoprotein subclass distribution during late pregnancy. AB - Plasma lipoprotein distribution during late pregnancy is unusual since high density lipoprotein (HDL) levels are increased in the presence of hypertriglyceridemia; the latter is usually associated with decreases in HDL levels. To determine whether there is a relationship between late-pregnancy lipid levels and specific HDL subclasses, HDL size distribution was determined by nondenaturing gradient gel electrophoresis (GGE) in a group of 36 women at 35 to 36 weeks of gestation and again at 6 weeks' postpartum, and in a group of 10 nonpregnant women. At 35 to 36 weeks of gestation, plasma triglyceride (TG) and cholesterol concentrations were significantly increased over postpartum levels (218 +/- 62 v 112 +/- 69 mg/dL and 234 +/- 48 v 197 +/- 36 mg/dL, respectively). During late pregnancy, apolipoprotein A-I (apo A-I) and HDL cholesterol concentrations were also increased relative to postpartum levels (211 +/- 42 v 168 +/- 20 mg/dL and 63 +/- 13 v 53 +/- 11 mg/dL, respectively). GGE analysis indicated that at 35 to 36 weeks of gestation, 86% of the subjects had a substantial increase of the most buoyant and largest of the HDL species, HDL2b; postpartum and nonpregnant HDL subclass distribution was characterized by the predominance of HDL3a, which are smaller, more dense HDL. The shift in the HDL subclass distribution during late pregnancy was associated with significant positive correlations between HDL2b and apo A-I (r = .50, P < .05) and HDL cholesterol (r = .60, P < .001). There were significant elevations in the concentrations of cholesteryl ester transfer protein (CETP) and estrogen during late pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246776 TI - Cephalic-phase insulin in obese and normal-weight men: relation to postprandial insulin. AB - Cephalic-phase insulin release (CPIR) and its relation to postprandial insulin release were examined in 18 normal-weight and 15 obese men. When the insulin data were expressed as absolute differences from baseline values, obese subjects exhibited significantly greater CPIR than normal-weight subjects (normals, 8.7 +/ 2.1 microU/mL/10 min; obese, 13.4 +/- 4.3 microU/mL/10 min; P < .01). Obese subjects were then separated into groups depending on their fasting insulin levels. This showed that only those subjects with elevated fasting insulin levels exhibited greater CPIR than normal subjects, and suggested that previous reports of exaggerated CPIR in the obese are merely a reflection of a basal hypersecretion of insulin. However, when insulin values were expressed as percentages of baseline, no significant differences between normal-weight and obese subjects were found, although a trend toward an attenuated response was observed in the obese group as a whole (normals, 81.6 +/- 19.1 microU/mL/10 min; obese, 51.3 +/- 16.1 microU/mL/10 min). A significant correlation between cephalic-phase and postprandial insulin release was found in normal-weight subjects (r = .62, P < .05), but not in obese subjects (r = .02, P < .9). PMID- 8246777 TI - Skeletal muscle glucose uptake during short-term contractile activity in vivo: effect of prior contractions. AB - The goal of the present study was to examine the time course of skeletal muscle glucose uptake and changes in intracellular metabolites occurring with the onset of in situ stimulation, and to assess the effect of a prior period of contractions on subsequent contraction-induced increases in glucose uptake. Hindlimb muscle in anesthetized rabbits was studied noninvasively using the positron-emitting glucose analog 18F-2-fluoro-deoxy-D-glucose (FDG). Fractional rates of FDG phosphorylation were measured on a minute-to-minute basis during rest, 3.5 minutes of priming exercise (PE), 15 or 30 minutes of PE recovery, and a subsequent 15-minute period of contractions. Muscles were electrically stimulated at 2 Hz, and force production was held constant during the contraction period(s). FDG uptake did not differ from control values either during PE or during 60 minutes of recovery from PE. In response to 15 minutes of contractions, muscle stimulated without PE demonstrated increased FDG uptake, but only after a delay of 5.0 +/- 0.7 minutes. Muscle with PE but rested 15 minutes had increased FDG uptake with a delay of 0.5 +/- 0.2 minutes, and muscle with PE but rested 30 minutes had increased FDG uptake after a delay of 8.0 +/- 0.9 minutes (P < .01 all groups). All groups reached similar levels of FDG uptake by the end of 15 minutes of contractions. Both groups with PE had control levels of adenosine triphosphate (ATP), phosphocreatine (PCr), and glucose-6-phosphate (G6P) after PE recovery, but glycogen level was lower than the control value (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246778 TI - Regulation of the growth hormone (GH) receptor and GH-binding protein by GH pulsatility. AB - Growth hormone (GH) secretion is pulsatile in man and every other mammalian species that has been studied. The magnitude of pulses, their frequency, and their regularity vary. The receptor, on its part, undergoes cycles of internalization and recycling that are in synchrony with the frequency of GH pulses. This sequence of events has been shown to be advantageous to growth. It is suggested that changes in GH secretion dynamics secondarily lead to most of the changes in GH receptor abundance and GH-binding protein (GH-BP) abundance. Across a wide scope of comparative studies, ontogenesis data, experimental systems, physiologic conditions, nutritional states, and disease situations, the pulsatility of serum GH is negatively correlated with cell membrane GH receptors and serum levels of GH-BP. It is suggested that these conditions regulate primarily the pattern of GH pulsatility, which in turn regulates the GH receptor/GH-BP, and thereby exert the specific effects on target cells to promote or suppress growth or to express distinct metabolic actions. PMID- 8246779 TI - Introduction to confocal microscopy and three-dimensional reconstruction. PMID- 8246780 TI - Measurement of intracellular pH with a laser scanning confocal microscope. PMID- 8246781 TI - Confocal microscopy of potentiometric fluorescent dyes. PMID- 8246782 TI - Imaging endoplasmic reticulum in living sea urchin eggs. PMID- 8246783 TI - Membrane glycolipid trafficking in living, polarized pancreatic acinar cells: assessment by confocal microscopy. PMID- 8246784 TI - Confocal immunofluorescence microscopy of microtubules in amphibian oocytes and eggs. PMID- 8246785 TI - Resolution of subcellular detail in thick tissue sections: immunohistochemical preparation and fluorescence confocal microscopy. PMID- 8246786 TI - Using confocal laser scanning microscopy to investigate the organization and development of neuronal projections labeled with DiI. PMID- 8246787 TI - Rapid scanning confocal microscopy. PMID- 8246788 TI - Confocal microscopy: important considerations for accurate imaging. PMID- 8246789 TI - Multicolor laser scanning confocal immunofluorescence microscopy: practical application and limitations. PMID- 8246790 TI - In vitro methods for measuring force and velocity of the actin-myosin interaction using purified proteins. PMID- 8246791 TI - Preparation of marked microtubules for the assay of the polarity of microtubule based motors by fluorescence microscopy. PMID- 8246792 TI - Expression of microtubule motor proteins in bacteria for characterization in in vitro motility assays. PMID- 8246793 TI - Tracking nanometer movements of single motor molecules. PMID- 8246794 TI - Assay of microtubule movement driven by single kinesin molecules. PMID- 8246795 TI - In vitro motility assays using microtubules tethered to Tetrahymena pellicles. PMID- 8246796 TI - Use of ATP analogs in motor assays. PMID- 8246797 TI - Myosin-mediated vesicular transport in the extruded cytoplasm of characean algae cells. PMID- 8246798 TI - Assay of membrane motility in interphase and metaphase Xenopus extracts. PMID- 8246799 TI - Microtubule motor-dependent formation of tubulovesicular networks from endoplasmic reticulum and Golgi membranes. PMID- 8246800 TI - Myosin-specific adaptations of the motility assay. PMID- 8246801 TI - Cytoplasmic extracts from the eggs of sea urchins and clams for the study of microtubule-associated motility and bundling. PMID- 8246802 TI - Kinesin-mediated vesicular transport in a biochemically defined assay. AB - Here we have described simple and reproducible methods to observe kinesin mediated vesicle and microtubule movements under defined conditions using video microscopy. We are optimistic that this assay will provide a useful tool to study kinesin function, regulation, and dynamic physical interactions with membranous organelles and microtubules. PMID- 8246803 TI - An assay for the activity of microtubule-based motors on the kinetochores of isolated Chinese hamster ovary chromosomes. PMID- 8246804 TI - The diatom central spindle as a model system for studying antiparallel microtubule interactions during spindle elongation in vitro. PMID- 8246805 TI - Motility of myosin I on planar lipid surfaces. PMID- 8246806 TI - Microtubule and axoneme gliding assays for force production by microtubule motor proteins. PMID- 8246807 TI - Analyzing microtubule motors in real time. PMID- 8246808 TI - Assays of axonemal dynein-driven motility. PMID- 8246809 TI - Evaluation of the opportunistic microbial flora and of some antimicrobial factors in the oral cavity of leukaemic patients. AB - The occurrence of opportunistic pathogens and the concentration of some antimicrobial factors in the oral cavity of both acute and chronic leukaemia patients were studied. Enterobacteria were isolated from both dental plaque and crevicular fluid of all the groups examined, with few differences between healthy volunteers and leukaemic subjects; yeasts were found in both the crevicular fluid and the dental plaque samples of chronic leukaemia patients, but only in the plaque of healthy volunteers. Acute leukaemia patients did not have yeasts, but they were the only group colonized by the pseudomonads. IgA and N-acetyl-D glucosaminidase (NAGase) significantly increased in chronic leukaemia patients compared with controls, whilst lysozyme seemed to present no marked differences for all groups. A further increase in NAGase concentration and an elevation in lysozyme content of saliva was observed for chronic leukaemia patients with severe periodontal lesions. PMID- 8246810 TI - Investigations into the growth of Mycobacterium leprae in a medium with palmitic acid under different gaseous environments. AB - Low oxygen tension has often been considered important for the growth of Mycobacterium leprae. Palmitic acid has been suggested as the oxidizable substrate for the in vitro cultivation of leprosy bacilli. The combined effects of palmitic acid and various known gas mixtures on the in vitro growth of M. leprae were investigated. When palmitic acid was included in the medium an optimal growth in both liquid and solid media was obtained between 16 to 20 weeks of incubation under gas mixtures containing 2.5 or 5% O2 and 5 or 10% CO2 as well as air. The use of different gas mixtures is tedious, time consuming and laborious. Since the cultures incubated under air gave the same cell yield as obtained when incubated under optimal gas mixtures, air alone can be used for the in vitro cultivation trials of M. leprae when palmitic acid is included in the culture medium. PMID- 8246811 TI - Studies on mutational cross-resistance between ciprofloxacin, novobiocin and coumermycin in Escherichia coli and Staphylococcus warneri. AB - Nalidixic acid resistant mutants of Escherichia coli KL16 were tested against ciprofloxacin, coumermycin and novobiocin. The mutants gyrA, nalB and nal-24 were more resistant than KL16 to ciprofloxacin, whereas the nal-31 strain was hypersensitive. Only the nalB mutant was more resistant to novobiocin than KL16, but gyrA, nal-31 and nal-24 mutants were more sensitive to coumermycin than KL16. Newly-isolated novobiocin-resistant mutants of KL16 were not cross-resistant to coumermycin or ciprofloxacin. Some coumermycin-resistant mutants were cross resistant to novobiocin but not ciprofloxacin, whereas mutants resistant to novobiocin and ciprofloxacin were isolated at higher coumermycin concentrations. Two types of Staphylococcus warneri mutant were isolated on media containing novobiocin or coumermycin. Each was resistant to both coumarins, but one was highly resistant to novobiocin and the other to coumermycin. High level resistance to both coumarins was unstable. E. coli mutants differed in susceptibility to bactericidal concentrations of ciprofloxacin, and S. warneri mutants behaved similarly. These results suggest the modes of action of the coumarins are not identical. PMID- 8246812 TI - Vasoactivity of isolated coronary arterial microvessels: influence of albumin. AB - Previous investigators have reported the development of spontaneous tone, a spontaneous decrease in diameter in response to application of intraluminal pressure, in isolated microvessels bathed in physiological salt solution (PSS) containing albumin. In contrast, myogenic activity and spontaneous tone are rare in microvessels bathed in albumin-free PSS. The purpose of this study was to determine whether albumin and/or the development of spontaneous tone altered coronary microvessel responsiveness to vasoactive agents. Microvessels (70-150 microns) were isolated from the hearts of domestic pigs and studied in vitro using video dimension analysis. Control microvessels were bathed in albumin-free PSS while albumin-treated vessels were bathed in PSS containing 10 mg/ml albumin. Vasoconstrictor responses were determined for acetylcholine (3 x 10(-10) to 1 x 10(-5) M) and KCl (20-120 mM). Concentration-response curves were generated for an endothelium-dependent vasodilator, bradykinin (1 x 10(-12) to 1 x 10(-7) M), and a direct smooth muscle dilator, sodium nitroprusside (1 x 10(-10) to 1 x 10( 4) M). Seventy-seven percent of the albumin-treated microvessels developed spontaneous tone, whereas only 20% of the control microvessels developed tone. KCl and acetylcholine induced contraction in both control and albumin-treated microvessels. Maximal response to acetylcholine (percentage constriction) was significantly greater in albumin-treated microvessels (79.93 +/- 2.53 and 68.86 +/- 3.20 in control and albumin-treated groups, respectively). Responses to bradykinin and sodium nitroprusside were not different between albumin-treated and control groups. The results indicate that microvessels develop spontaneous tone more frequently when bathed in PSS containing albumin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246813 TI - Capillary perfusion in the occluded-reperfused canine myocardium: evidence for slowed reflow. AB - The purpose of this study was to determine if the no-reflow phenomenon in the occluded/reperfused dog heart was anatomical (capillary bed could not be perfused) or functional (slowed capillary perfusion). The left anterior descending (LAD) coronary artery was isolated in anesthetized open-chest dogs. The LAD was occluded (2 hr) then reperfused (4 hr). A total of 14 dogs were assigned to one of two groups: The multiple pass (MP) or single pass (SP) group. Fluorescent labeled dextran (FITC) was injected and circulated either for 10 min in the MP group or for one pass of the circulation in the SP group. In the last 2 min, the MP group was asphyxiated. Hearts were excised and frozen in liquid nitrogen. The LAD and control region were isolated and frozen sections (5 microns) obtained. Sections were photographed under fluorescent light and then immunohistochemically stained. The microvessels were identified by visualizing the endothelium with a Factor VIII antibody. The specific areas photographed were relocated. The total number of capillaries, determined by stain outlining a lumen of no greater than 10 microns in diameter, were counted. This number was compared to the number of microvessels containing FITC-dextran. There was no difference in the total perfusable capillary bed in the MP group between the control and LAD regions (80 +/- 4 and 82 +/- 10%). In the SP group, there was a significant difference between the control and LAD regions (59 +/- 25 and 46 +/- 14%). The percentage of the capillary bed perfused in the SP group was less than the MP group in each region. Closest individual analysis of the perfused capillary distribution in the SP group, showed the distribution in the LAD region was random, potentially clustered and more dispersed compared to the control region. This study provides data against physical obstruction of the capillary bed secondary to ischemia. The data suggest sluggish and clustered perfusion in the occluded-reperfused myocardium during a single pass of the coronary circulation. PMID- 8246814 TI - Lymphatic capillary pressure in patients with primary lymphedema. AB - Flow and pressure dynamics in minute human lymphatics are unexplored. Lymphatic capillary pressure was measured by the servo-nulling technique at the foot dorsum of 14 patients with primary lymphedema and 15 healthy controls. Glass micropipettes (7-9 microns) were inserted under microscopic control into lymphatic microvessels previously stained by fluorescence microlymphography (FITC Dextran 150,000). Mean lymphatic capillary pressure was 7.9 +/- 3.4 mm Hg in the controls and 15.0 +/- 5.1 mm Hg in the patients. The difference was significant at the P < 0.001 level. In about half of the patients and control subjects studied pressure fluctuated by more than 3 mm Hg. The mean intralymphatic pressure of lymphedema patients was slightly below mean interstitial pressure measured by J. T. Christensen, N. J. Shaw, M. M. Hamas and H. K. Al Hassan (1985, Microcirc., Endothelium, Lymphatics 2, 267-384) (17.9 mm Hg) in lower leg lymphedema. Microlymphatic hypertension present in patients with primary lymphedema is probably an important factor for edema formation. PMID- 8246815 TI - A study of vasodilator responses evoked by antidromic stimulation of A delta afferent nerve fibers supplying normal and reinnervated rat skin. AB - Experiments have been carried out on anaesthetized rats to investigate some properties of the vasodilator responses evoked in skin by stimulation of small myelinated, A delta-fibers. Both normal and reinnervated skin in the saphenous nerve field were studied. In unoperated animals it was found that the magnitude and time course of the responses recorded from a particular patch of skin remain stable over a period of hours, but the responses varied in magnitude in different parts of the saphenous nerve territory. A delta-fiber evoked vasodilatation responses recorded from reinnervated skin were generally very similar to those seen in normal skin except that they tend to be smaller. These observations, taken together with previous findings, suggest that this ability of cutaneous A delta-fibers to influence the vasculature in the vicinity of their sensory endings has some physiological significance. Unfortunately, attempts to link these skin blood flow responses with activity of single, identified A delta-fiber units were not successful. PMID- 8246816 TI - Characterization of neuroblastoma xenograft in rat flank. I. Growth, interstitial fluid pressure, and interstitial fluid velocity distribution profiles. AB - The growth, interstitial fluid pressure (IFP) and interstitial fluid velocity (IFV) profiles of a human neuroblastoma propagated in the flank of an immune suppressed rat were characterized. IFP was measured in the tumor center as a function of size, while radial distributions of IFP and IFV were measured in 2-cm tumors. IFP and IFV were measured using the wick-in-needle and clearance of locally generated hydrogen techniques, respectively. These techniques have a high spatial resolution, permit repetitive measurements, and are minimally invasive. We observed that IFP in the neuroblastoma increased as the tumor grew. Furthermore, IFP increased and its IFV decreased from the periphery toward the center of the tumor. Measured IFP and IFV values were compared to theoretical expectations calculated from the Baxter and Jain mathematical model. The predictions were highly correlated to the measured IFP and IFV profiles with the transport impedence parameter alpha 2 = 24.4. From our measurement data and the Baxter-Jain equations, we computed the interstitium hydraulic conductivity for neuroblastoma to be 7.11 x 10(-6) cm2/mm Hg-sec. PMID- 8246817 TI - Dynamic changes in stress fiber expression in rat uterine vein endothelial cells associated with pregnancy. AB - En face endothelial preparations of rat uterine vein were stained with rhodamine phalloidin to investigate the dynamics of stress fiber expression during pregnancy. In prepregnant animals, somewhat plump, spindle-like endothelial cells of the uterine vein had only a few short stress fibers. With the progress of pregnancy, however, many long stress fibers appeared within the elongated endothelial cells. Within 2 hr after delivery, these stress fibers became dramatically decreased in number as the cells reverted from an elongated to a plump shape and returned to the prepregnancy level by 14 days postpartum. The uterine vein showed a significant increase in length during pregnancy and quickly shortened after delivery. Thus, expression of stress fibers in endothelial cells of the uterine vein seems to be related to the tension loaded on this vessel during its elongation in parallel with the marked growth of the uterine body during pregnancy. This study shows that stress fibers are dynamic structures that may serve to maintain endothelial cell integrity during the exertion of tensile stress on the vessel wall. PMID- 8246818 TI - Arteriolar bifurcation angles vary with position and when flow is changed. AB - Flow distribution at microvascular bifurcations is influenced by the geometry of the bifurcation region, an area which also is altered pathologically. Bifurcation geometry was measured by in vivo microscopy of the cremaster muscle of anesthetized (Nembutal, 70 mg/kg) golden hamsters (N = 40), at rest and during maximal dilation (10(-4) M adenosine). The sequential branches had progressively smaller angles of bifurcation at rest: (first position) 118 +/- 5 degrees; (second) 89 +/- 6 degrees; (third) 78 +/- 5 degrees; (last) 58 +/- 4 degrees. Between flow conditions, the angle at any bifurcation changed by up to +/- 50 degrees, and the angle change was related to position. For the first position, the angles that decreased vs those that increased were significantly different at rest (130 +/- 6 degrees vs 109 +/- 7 degrees), but not during maximal dilation (119 +/- 6 degrees vs 118 +/- 7 degrees). Conversely, at the last bifurcation, the resting angles were not different (58 +/- 5 degrees vs 56 +/- 7 degrees), but became significantly different during maximal dilation (48 +/- 6 degrees vs 68 +/ 6 degrees). The axial distance to the first branch ranged between 57 and 857 microns; the angle at the first position was significantly smaller for those first branches that arose further distally along the feed. Further, angles that decreased (vs those that increased) were from significantly longer transverse arterioles (total length: 1820 +/- 77 microns vs 1560 +/- 61 microns). Resting tone was related to the angle as the smaller angles at the first position, but not at the last position, were more constricted in diameter. Tone was differently related to angle change as the bifurcations that decreased (vs those that increased) in angle were significantly more constricted at the last position (branch diameter rest/maximal: 0.57 +/- 0.05 vs 0.81 +/- 0.08) but not at the first position (0.66 +/- 0.09 vs 0.64 +/- 0.05). Thus, we show that the angle of bifurcation varies systematically for sequential branches arising along a single transverse arteriole and that the angles change with flow. This systematic organization for the geometric shape of sequential bifurcation regions may participate in the regulation of flow distribution within this group of arterioles. PMID- 8246819 TI - Pathophysiological characteristics of cutaneous microcirculation in patients with liver cirrhosis: relationships to cardiovascular hemodynamics and plasma neurohormonal factors. AB - To elucidate pathophysiological characteristics of the peripheral circulation in liver cirrhosis, cutaneous microcirculation was analyzed at the finger, palm, toe, and arch of the foot using laser Doppler spectroscopy in 19 patients with liver cirrhosis, and the results were correlated with cardiovascular hemodynamics (n = 10) and plasma neurohormonal factors (n = 8). Cutaneous blood flow at each area was all significantly reduced (P < 0.001, < 0.05, < 0.01, < 0.001, respectively) in patients with liver cirrhosis compared to those in normals (n = 20). Cutaneous blood mass was also significantly reduced (P < 0.05, < 0.01, < 0.001, respectively) except at the arch of foot in patients with liver cirrhosis. There were significant correlations between finger cutaneous blood flow and systemic vascular resistance (r = -0.73, P < 0.05), plasma norepinephrine level (r = -0.84, P < 0.01), plasma renin activity (r = -0.77, P < 0.01), plasma concentrations of aldosterone (r = -0.76, P < 0.05) and angiotensin II (r = 0.76, P < 0.05), between palm blood flow and plasma vasoactive intestinal polypeptide concentration (r = 0.83, P < 0.05). From these results and our previous data demonstrating increased forearm muscular blood flow in patients with liver cirrhosis, increases in arteriovenous anastomotic flow and the contribution of neurohormonal factors will represent pathophysiological mechanisms for these changes. PMID- 8246820 TI - Fluorescence ratio imaging measurement of pH gradients: calibration and application in normal and tumor tissues. AB - The fluorescence ratio imaging technique is currently applied in vitro and in vivo to measure intracellular as well as extracellular pH. To date, however, no effort has been made to assess the technique's ability to detect spatial pH gradients in vitro or in vivo, where pH gradients are known to exist, in a controlled environment using an independent pH measurement system. We present here both in vitro calibration and in vivo application of fluorescence ratio imaging to measure pH gradients in a biologically useful pH range (6.2-7.8). Isoelectric focusing in short polyacrylamide gels was used to establish spatial pH gradients ranging from approximately 0.2 to 0.5 pH units/mm. The pH gradients were measured independently with fluorescence ratio imaging microscopy (FRIM) and a surface electrode. The average difference between measurements using the two techniques was 0.043 pH units for 10-mm polyacrylamide gels (100 measurements in 12 gels) and 0.046 pH units for 20-mm gels (114 measurements in 15 gels). These gel studies have demonstrated the use of isoelectric focusing to calibrate FRIM for measuring spatial pH gradients characteristic of those found in tumors in vivo. The feasibility of FRIM was tested in vivo by measuring pH in a solid tumor (VX2 carcinoma) and surrounding normal granulation tissue grown in the rabbit ear chamber. The average (+/- SD) pH of normal and neoplastic regions of the composite tissue was 7.18 +/- 0.11 (n = 158) and 6.75 +/- 0.10 (n = 191), respectively. The pH varied by an average of approximately 0.6 pH units over the 5-mm diameter of the chamber and showed a sharp difference at the interface of the two tissue types. These pH values are consistent with the current pH data on normal and neoplastic tissues. Thus FRIM should permit noninvasive in vivo evaluation of new pH-modifying agents and, in combination with other optical techniques, should offer novel mechanistic information about tumor pathophysiology. PMID- 8246821 TI - Measurement of material extravasation in microvascular networks using fluorescence video-microscopy. AB - We have developed a new method using fluorescence videomicroscopy to quantitate the extravasation of intravenously injected materials. This method can measure the relative plasma concentration of, and the vascular permeability to, these materials in microcirculatory preparations which contain multiple blood vessels in a field of view. The image of a tissue area containing multiple blood vessels is recorded via a SIT camera immediately before, and for an extended period after, the intravenous injection of a bolus of fluorescent test tracers. The videotape is analyzed off-line. At various time points, the light intensities of the entire tissue area and of several spots over selected vessels are measured. These measurements are then used to calculate the fluorescent light intensities arising from the tracers inside vessels (Iv) and in the interstitial region (Ii). Iv represents the relative amount of the tracers in the plasma, and Ii represents that in the interstitium. Iv and Ii are used to calculate an average permeability (P) for the vessels in the observed region. The benefit of this method is that it can be used to compare permeability of various tissues of interest or to serially evaluate changes in P in the same tissue over time. In this study, it was applied to measuring P to albumin as well as to liposomes in granulating and implanted tumor tissues in a rat skin flap window chamber. Changes in permeability to a small molecule (sulforhodamine B) before and during bradykinin application were also measured. The results of these experiments indicate that the relative plasma concentrations predicted by this method conformed well to those measured directly from blood samples, and the measured permeability values were consistent with previously published data. Therefore, this method provides a valid approach for quantitatively measuring the extravasation of intravenously injected molecular and colloidal materials in microcirculatory preparations. The method has a set of defined experimental conditions and assumptions that cannot be violated, however, or erroneous results can be obtained. PMID- 8246822 TI - Vero toxins (Shiga-like toxins) produced by enterohemorrhagic Escherichia coli (verocytotoxin-producing E. coli). PMID- 8246823 TI - The gene encoding the heat-stable enterotoxin of Vibrio cholerae is flanked by 123-base pair direct repeats. AB - The heat-stable enterotoxin (O1-ST) gene (sto) was cloned from chromosome of the strain GP156 of Vibrio cholerae O1 (Inaba, El Tor) in Escherichia coli K-12, and its nucleotide sequence was determined. The nucleotide sequence of sto was very similar to that of NAG-ST gene (stn) of V. cholerae non-O1. Both sto and stn were flanked by 123-base pair direct repeats which had at least 93% homology to one another and included some inverted repeats. All the strains of V. cholerae, V. mimicus, V. metschnikovii, V. hollisae and Yersinia enterocolitica examined by colony hybridization had the direct repeat sequence regardless of ST-gene possession. PMID- 8246824 TI - Comparison of detection methods for Legionella species in environmental water by colony isolation, fluorescent antibody staining, and polymerase chain reaction. AB - Three detection methods for Legionella species in water samples from cooling towers and a river were examined. Direct counting of bacteria stained with fluorescent antibody (FA) for L. pneumophila (serogroups 1 to 6) could detect the cell of 10(4) to 10(6) cell/100 ml in all 14 samples, while colony counting method detected 10 to 10(3) CFU/100 ml only in 8 samples from cooling towers. Polymerase chain reaction (PCR) assay with primers to amplify 16S ribosomal DNA sequence of most Legionella species (LEG primer) detected legionellae in 13 samples, while species-specific primers for L. pneumophila detected the DNAs from 3 samples. In laboratory examination, LEG primers could amplify DNAs of 29 species of genus Legionella with high sensitivity, even from 1 cell of L. pneumophila GIFU 9134. The PCR assay with LEG primers was specific and sensitive methods to be satisfied the survey of legionellae. Thus, PCR assay is a suitable method to detect and monitor Legionella species in an environment. PMID- 8246825 TI - Heterogeneity of HLA-G genes identified by polymerase chain reaction/single strand conformational polymorphism (PCR/SSCP). AB - A genomic HLA-G clone named 7.0E was isolated from a Japanese placenta. The deduced amino acid sequence of the 7.0E was identical to two HLA-G genomic clones and two cDNA clones previously described. The DNA sequences of alpha 1 and alpha 2 domains of the HLA-G gene from 5 cell lines also encoded the same amino acids. However, a 14 bp insertion, ATTTGTTCATGCCT, was present in the 3' untranslated region of 7.0E compared with the originally described HLA-G clone (HLA 6.0). Polymerase chain reaction (PCR)/single strand conformational polymorphism (SSCP) analysis of exon 8 allowed the HLA-G gene to be classified into two alternative types, G6.0 and 7.0 E, those correlated to the absence or the presence of the 14 bp stretch. Each group had minor sequence variant(s), and the alleles of the 7.0E type were more heterogeneous than those of the G6.0-type. The 14 bp deletion is present only in the G6.0-type of HLA-G alleles among HLA class I genes. Thus it was suggested that G6.0 alleles were generated after diversification of the HLA G. PMID- 8246826 TI - Radioresistance of intermediate TCR cells and their localization in the body of mice revealed by irradiation. AB - Extrathymic generation of T cells in the liver and in the intestine was recently demonstrated. We investigated herein whether such T cells, especially those in the liver, are present in other organs of mice. This investigation is possible employing our recently introduced method with which even a minor proportion of extrathymic, intermediate TCR cells in organs other than the liver can be identified. Intermediate TCR cells expressed higher levels of IL-2R beta and LFA 1 than bright TCR cells (i.e., T cells of thymic origin) as revealed by two-color staining. Although intermediate TCR cells were present at a small proportion in the spleen and thymus, they predominated in these organs after irradiation (9 Gy) and bone marrow reconstitution, or after low dose irradiation (6 Gy). This was due to that intermediate TCR cells were relatively radioresistant, whereas bright TCR cells were radiosensitive. Microscopic observation and immunochemical staining showed that intermediate TCR cells in the spleen localized in the red pulp and those in the thymus localized in the medulla. These intermediate TCR cells displayed a large light scatter, similar to such cells in the liver. The present results suggest that intermediate TCR cells may proliferate at multiple sites in the body. PMID- 8246827 TI - A comparative study on T cell receptor V beta gene usages: spleen cells from the non-obese diabetic (NOD) mouse and its non-diabetic sister strain, the ILI mouse, and infiltrating T cells into pancreata of NOD mice. AB - We analyzed the usage of T cell receptor (TCR) V beta genes of spleen cells of NOD mice in comparison with those of its non-diabetic sister strain ILI mice which show no insulitis and (ILI x NOD)F1 mice. The quantitative polymerase chain reaction (PCR) method revealed that PCR V beta repertoires of these mice are indistinguishable. This is consistent with our previous observation that ILI mice share the same H-2 class II genes with NOD mice. PCR method also revealed that the V beta transcript of infiltrating T cells into pancreas of NOD mice was not restricted but was rather diverse. The role of TCR repertoire in the development of insulitis was discussed. PMID- 8246828 TI - Host-specific hemagglutination of influenza A (H1N1) virus. AB - H1N1 strains of influenza A virus isolated during the influenza season of 1991-92 were divided into two groups according to the property of host-specific hemagglutination. Group 1 viruses agglutinated human and chicken red blood cells. Group 2 viruses agglutinated human but not chicken red blood cells. The viruses of both groups, however, showed the same antigenic structure determined with ferret antisera. The virus clones which were plaque-purified twice from a group 2 virus retained the characteristic of host-specific hemagglutination after five successive passages in MDCK cells, indicating that this phenomenon is genetically determined. However, the amino acid sequences of the hemagglutinin (HA) polypeptides deduced from the nucleotide sequences of the HA gene of the two groups did not show any differences between them. This suggests a difference in amino acids in some other polypeptide(s), which affects the host-specific hemagglutination. PMID- 8246829 TI - Release of prostaglandin E2 by human mononuclear cells exposed to heat-killed Salmonella typhi. AB - Human mononuclear cells pre-labeled with [3H]arachidonic acid were shown to release metabolites following in vitro addition of heat-killed Salmonella typhi (HKST). The amount of label released was significantly higher than that seen with live S. typhi (LST). Addition of increasing amounts of HKST resulted in an increased release of metabolites. Enzyme immunoassay of the culture supernatants revealed that the bulk of the metabolite released was prostaglandin E2 (PGE2). Leukotriene B4 (LTB4) and leukotriene C4 (LTC4) were not detectable in the culture supernatants. The significance and implications of these results are discussed. PMID- 8246830 TI - Drugs for parasitic infections. PMID- 8246831 TI - Loss of the basement membrane matrix molecule, bamin, in diphenylamine-treated mice. AB - Polycystic kidney disease (PKD) is a life-threatening disease characterized by focal dilatations or cysts in certain kidney tubules. Changes (i.e. thickening) in the support structure for these tubules, the basement membrane, have been related to the development of the cysts. Analysis of changes in basement membranes of humans with PKD is difficult, however, due to the restricted amount of material available for study. Several genetic and induced animal models, including diphenylamine-treated rats, have been employed to study the effects of PKD on basement membrane synthesis. While all these studies agree that PKD has a significant influence on basement membranes, no clear understanding as to how PKD effects basement membrane composition has emerged. Here, we report our findings of the effect of diphenylamine treatment on the composition of the basement membrane. Our immunohistological studies indicate that bamin, a recently described glycoprotein associated with glomerular basement membranes (Robinson et al., 1989), is not present in the glomerular basement membranes of diphenylamine treated mice. This finding was confirmed by analysis of the composition of the basement membrane matrix synthesized by EHS tumors grown in control and diphenylamine-treated mice. The possible role of bamin in the pathogenesis of renal cysts is discussed. PMID- 8246832 TI - Glycosaminoglycans and chondroitin/dermatan sulfate proteoglycans in the myocardium of a non-human primate. AB - Lyophilized mid-wall left ventricular myocardial tissues of the long-tailed non human primate, Macaca fascicularis, were examined for the presence of glycosaminoglycans and chondroitin/dermatan sulfate proteoglycans. Mean uronic acid concentration in all samples was 0.97 +/- 0.27 micrograms per mg dry weight myocardium. The distribution of the glycosaminoglycans in the myocardium, determined by cellulose acetate strip electrophoresis was 62 +/- 4% heparan sulfate, 20 +/- 6% hyaluronan, and 16 +/- 5% chondroitin/dermatan sulfate. The analysis of chondroitin/dermatan sulfate proteoglycans, done directly on the extracts of lyophilized myocardium using agarose-acrylamide gel electrophoresis and Western blotting with monoclonal antibodies to various carbohydrate epitopes and with polyclonal antibodies to the protein core, showed the presence of biglycan and decorin. That these two and no other chondroitin/dermatan sulfates were present was established by core protein analysis using SDS PAGE and Western blotting. Quantification of chondroitin/dermatan sulfate proteoglycans uncovered high individual specific variability of the chondroitin/dermatan sulfate epitopes, but only moderate variability of biglycan and decorin core proteins. The variability of the chondroitin/dermatan sulfate epitopes is most likely related to individual specific differences in chain number, iduronate content and sulfation patterns of biglycan and decorin. PMID- 8246833 TI - Effect of aging and dietary restriction on matrix Gla protein and other components of rat tracheal cartilage. AB - Age-related changes in connective tissues can alter their functions of elasticity, compressibility and support. Matrix Gla protein (MGP) is a vitamin K dependent connective tissue component of unknown function. We have purified bovine MGP, and developed a specific radioimmunoassay for it. Since it is found in highest concentration in cartilage, we have developed quantitative extraction methods for MGP, and examined the age-related changes of MGP relative to other components found in the cartilage matrix. The ratio of hydroxyproline to MGP increases with age, while the ratio of glycosaminoglycan to MGP is constant. No effect is seen for MGP in the dietary restricted rat with prolonged lifespan, while both hydroxyproline and glycosaminoglycan contents of tracheal cartilage are significantly increased by dietary restriction (p < or = .05). These data show that MGP and glycosaminoglycan concentration are relatively constant in rats from 6 to 30 months of age, while hydroxyproline concentration increases with age. PMID- 8246834 TI - Interleukin-1 alpha and epidermal growth factor synergistically enhance the release of collagenase by periosteal connective tissue in vitro. AB - The effects of recombinant human interleukin-1 alpha (IL-1 alpha) and murine epidermal growth factor (EGF) on the release of collagenase were studied in an in vitro model system using periosteal explants from rabbit calvariae. Following an incubation period of 72 h it was shown that IL-1 alpha in combination with EGF (IL-1 alpha + EGF) induced a synergistic increase in the amount of collagenase released by periosteal explants. This increase appeared to be at least 10-fold. Most of the enzyme was present in a latent form since the increase in enzyme activity was only detectable after activation by APMA and the molecular weight as determined in immunoblots corresponded to the latent form of this enzyme. Incubations carried out with IL-1 alpha alone resulted in a 2- to 4-fold increase of total enzyme activity, whereas the amount of collagenase in media of EGF treated periosteal did not surpass control values. A neutralizing anti-IL-1 alpha antibody completely blocked the enhanced release of collagenase as induced both by IL-1 alpha and by IL-1 alpha + EGF. Indomethacin partially prevented the IL-1 alpha + EGF-induced increase in enzyme release, suggesting the involvement of prostaglandins. The amount of tissue inhibitor of metalloproteinases (TIMP) as determined by ELISA was slightly elevated in culture media obtained from all cytokine-treated explants. Comparable results were obtained by Western blot analysis as well as by a functional bioassay. It is suggested that the concomitant presence of the cytokines IL-1 alpha and EGF may play an important role in collagenase-mediated degradation of collagen. PMID- 8246835 TI - Expression of human COL1A1 gene in stably transfected HT1080 cells: the production of a thermostable homotrimer of type I collagen in a recombinant system. AB - A recombinant system was developed for the production of homotrimeric type I collagen in stably transfected HT1080 cells. A DNA construct (COL1A1-CMV) was prepared that contained the cDNA for the human COL1A1 gene under the transcriptional control of the promoter and enhancer of the immediate early gene of CMV. The construct, which also contained a neomycin-resistance gene, was transfected into HT1080 cells, a human fibrosarcoma cell line that synthesizes type IV collagen but does not normally synthesize any of the fibrillar collagens. Cells derived from the neomycin-resistant transfectants were then screened using a polyclonal antibody specific for human pro alpha 1(I) chains in order to identify clones that secreted high levels of the pro alpha(I) chain of type I procollagen. About 2% of neomycin-resistant clones secreted procollagen that consisted of a homotrimer of pro alpha 1(I) chains. The procollagen was post translationally over-modified as judged by slower migration on SDS-polyacrylamide gel electrophoresis of the pro alpha 1(I) chains compared to pro alpha 1(I) chains of normal type I procollagen. The procollagen was triple helical as assayed by protease digestion with a variable cleavage at 38 degrees C and a thermal transition of both the intact and partially cleaved protein of about 41 degrees C. The system provides a method of expressing genes for fibrillar procollagens so that fully recombinant proteins are generated and easily isolated. PMID- 8246836 TI - Collagen fibril formation in the presence of alkyl sulfonate detergents. AB - The effects of sulfonate detergents on fibril formation by type I (rat tail tendon) collagen, untreated and pepsin-treated, were studied at two temperatures. Fluorescent probes and CD spectra were used to study possible conformational changes in the collagen. With untreated collagen, the rate of fibril formation was reduced by the presence of the detergents, particularly at the low temperature and when the detergent molecules had longer aliphatic chains. With pepsin-treated collagen, however, the rate was increased by the longer-chain detergent at the higher temperature. These observations are discussed in the light of present knowledge of the role of telopeptides and hydrophobic interactions in fibril formation. PMID- 8246837 TI - Dissimilar expression patterns for the extracellular matrix proteins osteopontin (OPN) and collagen type I in dental tissues and alveolar bone of the neonatal rat. AB - Osteopontin (OPN) is a phosphorylated, sialic acid containing glycoprotein that can be extracted from the mineralized extracellular matrix of bone. In the present study we determined the expression patterns of OPN in dental tissues and alveolar bone of 1-3 day old (neonatal) rats by means of 1) immunohistochemistry, 2) Northern blotting and 3) in situ hybridization. We compared these patterns with those of type I collagen. We localized collagen type I expression in osteoblasts adjacent to alveolar bone and in odontoblasts lining predentin/dentin, but not in the epithelial ameloblasts. For OPN, we observed a weak antigenicity in predentin. Although generally no cellular immunostaining was found, very occasionally a minor immunoreactivity was detected in a small number of pre-mineralizing incisor odontoblasts. On the mRNA level, however, no OPN transcripts could be detected in odontoblasts, either by in situ or by Northern hybridization analyses. Also the odontoblasts of the bone-like dentin (osteodentin) region in the tip of incisors were negative for OPN. In contrast, however, osteoblasts of alveolar bone showed strong positive signals with all three techniques, confirming the sensitivity and specificity of the detection methods. From the data obtained in this study, it can be concluded that during early stages of dentinogenesis OPN presumably is not expressed in developing rat tooth germs. The weak immunostaining observed sporadically in some young odontoblasts is probably due to resorption of OPN of non-dental origin entrapped in the predentin. PMID- 8246838 TI - The Second International Workshop on Scleroderma Research. PMID- 8246839 TI - Regulation of fatty acid biosynthesis in Escherichia coli. AB - Our understanding of fatty acid biosynthesis in Escherichia coli has increased greatly in recent years. Since the discovery that the intermediates of fatty acid biosynthesis are bound to the heat-stable protein cofactor termed acyl carrier protein, the fatty acid synthesis pathway of E. coli has been studied in some detail. Interestingly, many advances in the field have aided in the discovery of analogous systems in other organisms. In fact, E. coli has provided a paradigm of predictive value for the synthesis of fatty acids in bacteria and plants and the synthesis of bacterial polyketide antibiotics. In this review, we concentrate on four major areas of research. First, the reactions in fatty acid biosynthesis and the proteins catalyzing these reactions are discussed in detail. The genes encoding many of these proteins have been cloned, and characterization of these genes has led to a better understanding of the pathway. Second, the function and role of the two essential cofactors in fatty acid synthesis, coenzyme A and acyl carrier protein, are addressed. Finally, the steps governing the spectrum of products produced in synthesis and alternative destinations, other than membrane phospholipids, for fatty acids in E. coli are described. Throughout the review, the contribution of each portion of the pathway to the global regulation of synthesis is examined. In no other organism is the bulk of knowledge regarding fatty acid metabolism so great; however, questions still remain to be answered. Pursuing such questions should reveal additional regulatory mechanisms of fatty acid synthesis and, hopefully, the role of fatty acid synthesis and other cellular processes in the global control of cellular growth. PMID- 8246841 TI - Trichothecene biosynthesis in Fusarium species: chemistry, genetics, and significance. AB - Several species of the genus Fusarium and related fungi produce trichothecenes which are sesquiterpenoid epoxides that act as potent inhibitors of eukaryotic protein synthesis. Interest in the trichothecenes is due primarily to their widespread contamination of agricultural commodities and their adverse effects on human and animal health. In this review, we describe the trichothecene biosynthetic pathway in Fusarium species and discuss genetic evidence that several trichothecene biosynthetic genes are organized in a gene cluster. Trichothecenes are highly toxic to a wide range of eukaryotes, but their specific function, if any, in the survival of the fungi that produce them is not obvious. Trichothecene gene disruption experiments indicate that production of trichothecenes can enhance the severity of disease caused by Fusarium species on some plant hosts. Understanding the regulation and function of trichothecene biosynthesis may aid in development of new strategies for controlling their production in food and feed products. PMID- 8246842 TI - Molecular biology of the lignin-degrading basidiomycete Phanerochaete chrysosporium. AB - The white rot basidiomycete Phanerochaete chrysosporium completely degrades lignin and a variety of aromatic pollutants during the secondary metabolic phase of growth. Two families of secreted heme enzymes, lignin peroxidase (LiP) and manganese peroxidase (MnP), are major components of the extracellular lignin degradative system of this organism. MnP and LiP both are encoded by families of genes, and the lip genes appear to be clustered. The lip genes contain eight or nine short introns; the mnp genes contain six or seven short introns. The sequences surrounding active-site residues are conserved among LiP, MnP, cytochrome c peroxidase, and plant peroxidases. The eight LiP cysteine residues align with 8 of the 10 cysteines in MnP. LiPs are synthesized as preproenzymes with a 21-amino-acid signal sequence followed by a 6- or 7-amino-acid propeptide. MnPs have a 21- or 24-amino-acid signal sequence but apparently lack a propeptide. Both LiP and MnP are regulated at the mRNA level by nitrogen, and the various isozymes may be differentially regulated by carbon and nitrogen. MnP also is regulated at the level of gene transcription by Mn(II), the substrate for the enzyme, and by heat shock. The promoter regions of mnp genes contain multiple heat shock elements as well as sequences that are identical to the consensus metal regulatory elements found in mammalian metallothionein genes. DNA transformation systems have been developed for P. chrysosporium and are being used for studies on gene regulation and for gene replacement experiments. PMID- 8246843 TI - Colibri: a functional data base for the Escherichia coli genome. AB - Several data libraries have been created to organize all the data obtained worldwide about the Escherichia coli genome. Because the known data now amount to more than 40% of the whole genome sequence, it has become necessary to organize the data in such a way that appropriate procedures can associate knowledge produced by experiments about each gene to its position on the chromosome and its relation to other relevant genes, for example. In addition, global properties of genes, affected by the introduction of new entries, should be present as appropriate description fields. A data base, implemented on Macintosh by using the data base management system 4th Dimension, is described. It is constructed around a core constituted by known contigs of E. coli sequences and links data collected in general libraries (unmodified) to data associated with evolving knowledge (with modifiable fields). Biologically significant results obtained through the coupling of appropriate procedures (learning or statistical data analysis) are presented. The data base is available through a 4th Dimension runtime and through FTP on Internet. It has been regularly updated and will be systematically linked to other E. coli data bases (M. Kroger, R. Wahl, G. Schachtel, and P. Rice, Nucleic Acids Res. 20(Suppl.):2119-2144, 1992; K. E. Rudd, W. Miller, C. Werner, J. Ostell, C. Tolstoshev, and S. G. Satterfield, Nucleic Acids Res. 19:637-647, 1991) in the near future. PMID- 8246844 TI - Mechanisms of genome propagation and helper exploitation by satellite phage P4. AB - Temperate coliphage P2 and satellite phage P4 have icosahedral capsids and contractile tails with side tail fibers. Because P4 requires all the capsid, tail, and lysis genes (late genes) of P2, the genomes of these phages are in constant communication during P4 development. The P4 genome (11,624 bp) and the P2 genome (33.8 kb) share homologous cos sites of 55 bp which are essential for generating 19-bp cohesive ends but are otherwise dissimilar. P4 turns on the expression of helper phage late genes by two mechanisms: derepression of P2 prophage and transactivation of P2 late-gene promoters. P4 also exploits the morphopoietic pathway of P2 by controlling the capsid size to fit its smaller genome. The P4 sid gene product is responsible for capsid size determination, and the P2 capsid gene product, gpN, is used to build both sizes. The P2 capsid contains 420 capsid protein subunits, and P4 contains 240 subunits. The size reduction appears to involve a major change of the whole hexamer complex. The P4 particles are less stable to heat inactivation, unless their capsids are coated with a P4-encoded decoration protein (the psu gene product). P4 uses a small RNA molecule as its immunity factor. Expression of P4 replication functions is prevented by premature transcription termination effected by this small RNA molecule, which contains a sequence that is complementary to a sequence in the transcript that it terminates. PMID- 8246840 TI - Phosphoenolpyruvate:carbohydrate phosphotransferase systems of bacteria. AB - Numerous gram-negative and gram-positive bacteria take up carbohydrates through the phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS). This system transports and phosphorylates carbohydrates at the expense of PEP and is the subject of this review. The PTS consists of two general proteins, enzyme I and HPr, and a number of carbohydrate-specific enzymes, the enzymes II. PTS proteins are phosphoproteins in which the phospho group is attached to either a histidine residue or, in a number of cases, a cysteine residue. After phosphorylation of enzyme I by PEP, the phospho group is transferred to HPr. The enzymes II are required for the transport of the carbohydrates across the membrane and the transfer of the phospho group from phospho-HPr to the carbohydrates. Biochemical, structural, and molecular genetic studies have shown that the various enzymes II have the same basic structure. Each enzyme II consists of domains for specific functions, e.g., binding of the carbohydrate or phosphorylation. Each enzyme II complex can consist of one to four different polypeptides. The enzymes II can be placed into at least four classes on the basis of sequence similarity. The genetics of the PTS is complex, and the expression of PTS proteins is intricately regulated because of the central roles of these proteins in nutrient acquisition. In addition to classical induction repression mechanisms involving repressor and activator proteins, other types of regulation, such as antitermination, have been observed in some PTSs. Apart from their role in carbohydrate transport, PTS proteins are involved in chemotaxis toward PTS carbohydrates. Furthermore, the IIAGlc protein, part of the glucose specific PTS, is a central regulatory protein which in its nonphosphorylated form can bind to and inhibit several non-PTS uptake systems and thus prevent entry of inducers. In its phosphorylated form, P-IIAGlc is involved in the activation of adenylate cyclase and thus in the regulation of gene expression. By sensing the presence of PTS carbohydrates in the medium and adjusting the phosphorylation state of IIAGlc, cells can adapt quickly to changing conditions in the environment. In gram-positive bacteria, it has been demonstrated that HPr can be phosphorylated by ATP on a serine residue and this modification may perform a regulatory function. PMID- 8246845 TI - Genetics of eukaryotic RNA polymerases I, II, and III. AB - The transcription of nucleus-encoded genes in eukaryotes is performed by three distinct RNA polymerases termed I, II, and III, each of which is a complex enzyme composed of more than 10 subunits. The isolation of genes encoding subunits of eukaryotic RNA polymerases from a wide spectrum of organisms has confirmed previous biochemical and immunological data indicating that all three enzymes are closely related in structures that have been conserved in evolution. Each RNA polymerase is an enzyme complex composed of two large subunits that are homologous to the two largest subunits of prokaryotic RNA polymerases and are associated with smaller polypeptides, some of which are common to two or to all three eukaryotic enzymes. This remarkable conservation of structure most probably underlies a conservation of function and emphasizes the likelihood that information gained from the study of RNA polymerases from one organism will be applicable to others. The recent isolation of many mutations affecting the structure and/or function of eukaryotic and prokaryotic RNA polymerases now makes it feasible to begin integrating genetic and biochemical information from various species in order to develop a picture of these enzymes. The picture of eukaryotic RNA polymerases depicted in this article emphasizes the role(s) of different polypeptide regions in interaction with other subunits, cofactors, substrates, inhibitors, or accessory transcription factors, as well as the requirement for these interactions in transcription initiation, elongation, pausing, termination, and/or enzyme assembly. Most mutations described here have been isolated in eukaryotic organisms that have well-developed experimental genetic systems as well as amenable biochemistry, such as Saccharomyces cerevisiae, Drosophila melanogaster, and Caenorhabditis elegans. When relevant, mutations affecting regions of Escherichia coli RNA polymerase that are conserved among eukaryotes and prokaryotes are also presented. In addition to providing information about the structure and function of eukaryotic RNA polymerases, the study of mutations and of the pleiotropic phenotypes they imposed has underscored the central role played by these enzymes in many fundamental processes such as development and cellular differentiation. PMID- 8246848 TI - The coming crisis of public health in the suburbs. AB - A process of urban decay, coupled with forced displacement and behavioral problems, affecting an increasing number of neighborhoods in large U.S. cities. The resulting social disintegration has intensified a nexus of deviant behaviors and conditions, including substance abuse, that are related to transmission of HIV and resurgence of other contagious diseases. These diseases will diffuse, or are already diffusing, along the transportation hierarchy from larger into smaller central cities, and radially from the central cities into the surrounding areas. A widespread program of urban reform is a critical precondition for the control of contagious disease in the United States. It also is important for housing, social services, housing-related public services, and public health across urban and suburban jurisdictions. PMID- 8246846 TI - The phycobilisome, a light-harvesting complex responsive to environmental conditions. AB - Photosynthetic organisms can acclimate to their environment by changing many cellular processes, including the biosynthesis of the photosynthetic apparatus. In this article we discuss the phycobilisome, the light-harvesting apparatus of cyanobacteria and red algae. Unlike most light-harvesting antenna complexes, the phycobilisome is not an integral membrane complex but is attached to the surface of the photosynthetic membranes. It is composed of both the pigmented phycobiliproteins and the nonpigmented linker polypeptides; the former are important for absorbing light energy, while the latter are important for stability and assembly of the complex. The composition of the phycobilisome is very sensitive to a number of different environmental factors. Some of the filamentous cyanobacteria can alter the composition of the phycobilisome in response to the prevalent wavelengths of light in the environment. This process, called complementary chromatic adaptation, allows these organisms to efficiently utilize available light energy to drive photosynthetic electron transport and CO2 fixation. Under conditions of macronutrient limitation, many cyanobacteria degrade their phycobilisomes in a rapid and orderly fashion. Since the phycobilisome is an abundant component of the cell, its degradation may provide a substantial amount of nitrogen to nitrogen-limited cells. Furthermore, degradation of the phycobilisome during nutrient-limited growth may prevent photodamage that would occur if the cells were to absorb light under conditions of metabolic arrest. The interplay of various environmental parameters in determining the number of phycobilisomes and their structural characteristics and the ways in which these parameters control phycobilisome biosynthesis are fertile areas for investigation. PMID- 8246849 TI - Long-term care, Medicaid, and impoverishment of the elderly. AB - The 1989 Long Term Care Survey was used to assess the extent to which entry of disabled elderly into nursing homes depletes personal wealth ("spenddown"); the effect of state eligibility rules on this process, on incentives to shield assets for Medicaid eligibility, and on out-of-pocket prices for nursing-home care; and whether the disabled elderly's housing wealth represents potential revenue for Medicaid. Most disabled elderly in the community were either Medicaid eligible, or would have been soon after entering a nursing home. Variations in state Medicaid eligibility rules do not explain differences in the spenddown process: the amount of nonhousing wealth of most disabled elderly seldom warranted its transfer to relatives in order to secure Medicaid benefits, nor would their housing wealth have offered more than limited revenue for Medicaid. PMID- 8246850 TI - Is family care on the decline? A longitudinal investigation of the substitution of formal long-term care services for informal care. AB - The focus on rising costs of long-term care now encompasses community as well as institutional care. Policy makers cite the potential impact of changing social trends on informal caregivers' availability to continue as the main source of care and the possibility that formal services will then replace this informal care. They fear that families will relinquish their caregiving role if publicly funded home care services are available. Longitudinal data from a sample of disabled elders were used to investigate the substitution of formal services for informal care over a seven-year period. The substitution that was detected could be traced to the limited availability of informal care, and it represented a temporary change in the informal care pattern rather than a permanent replacement for it. Instead, use of formal services has supported the elderly person's continued residence in the community. PMID- 8246847 TI - Polypeptides of Treponema pallidum: progress toward understanding their structural, functional, and immunologic roles. Treponema Pallidum Polypeptide Research Group. AB - Treponema pallidum subsp. pallidum, the spirochete that causes syphilis, is unusual in a number of respects, including its small genome size, inability to grow under standard in vitro culture conditions, microaerophilism, apparent paucity of outer membrane proteins, structurally complex periplasmic flagella, and ability to evade the host immune responses and cause disease over a period of years to decades. Many of these attributes are related ultimately to its protein content. Our knowledge of the activities, structure, and immunogenicity of its proteins has been expanded by the application of recombinant DNA, hybridoma, and structural fractionation techniques. The purpose of this monograph is to summarize and correlate this new information by using two-dimensional gel electrophoresis, monoclonal antibody reactivity, sequence data, and other properties as the bases of polypeptide identification. The protein profiles of the T. pallidum subspecies causing syphilis, yaws, and endemic syphilis are virtually indistinguishable but differ considerably from those of other treponemal species. Among the most abundant polypeptides are a group of lipoproteins of unknown function that appear to be important in the immune response during syphilitic infection. The periplasmic flagella of T. pallidum and other spirochetes are unique with regard to their protein content and ultrastructure, as well as their periplasmic location. They are composed of three core proteins (homologous to the other members of the eubacterial flagellin family) and a single, unrelated sheath protein; the functional significance of this arrangement is not understood at present. Although the bacterium contains the chaperonins GroEL and DnaK, these proteins are not under the control of the heat shock regulon as they are in most organisms. Studies of the immunogenicity of T. pallidum proteins indicate that many may be useful for immunodiagnosis and immunoprotection. Future goals in T. pallidum polypeptide research include continued elucidation of their structural locations and functional activities, identification and characterization of the low-abundance outer membrane proteins, further study of the immunoprotective and immunodiagnostic potential of T. pallidum proteins, and clarification of the roles of treponemal proteins in pathogenesis. PMID- 8246851 TI - How state policy affects rural hospital consortia: the rural health care delivery system. AB - Rural hospitals face the issue of survival on a daily basis. Because of their small economic base and more complex service area, rural hospitals will require external assistance. The impact of government regulations on the formation of rural hospital consortia is scrutinized, particularly the incentives arising through Medicaid, certificate of need, and physician licensure requirements that spur hospitals to join consortia. These organizations of individual hospitals appear to be a feasible option for ensuring rural hospital survival. PMID- 8246852 TI - Evaluating ethics committees: a view from the outside. AB - More than 60 percent of hospitals have ethics committees. The wide-scale, voluntary adoption of these committees by hospitals is surprising, given the lack of data on their effectiveness. Little effort has been made to evaluate such committees, in part because of the lack of consensus on their purpose. The argument presented here is that, from a policy perspective, the committees' purpose must be to safe-guard patients' interests; a framework for evaluating committees based on that objective is outlined. The criteria for evaluation include access, quality, and cost effectiveness. Existing data used to assess ethics committees, using these criteria. Based on available data, it is questionable whether ethics committees are performing any better or worse than alternative mechanisms to achieve the same goal. PMID- 8246853 TI - Absence of reported measles--United States, November 1993. PMID- 8246854 TI - Infant mortality--United States, 1991. AB - The final infant (< 1 year of age) mortality rate for the United States for 1991 8.9 infant deaths per 1000 live births--was the lowest rate ever recorded and represented a decrease of 3% from the rate of 9.2 for 1990 (Figure 1). Based on provisional data, the trend of declining infant mortality continued through 1992 (rate: 8.5) (1). Infant mortality rates varied by race; race reflected differing distributions of several risk factors for infant death (e.g., low birthweight [LBW] [< 2500 g (5 lbs 9 oz) at birth]) and is useful for identifying groups at greatest risk for infant death. This report uses race-specific information from birth and death certificates compiled by CDC's National Center for Health Statistics' Viral Statistics System (2) to characterize infant mortality in 1991 and compares findings with those for 1990. PMID- 8246855 TI - Outbreaks of Mycoplasma pneumoniae respiratory infection--Ohio, Texas, and New York, 1993. AB - From June through November 1993, three outbreaks of acute respiratory illness (ARI) occurred in institutional settings in Ohio, Texas, and New York. This report summarizes investigations by state and local public health officials, military personnel, and CDC, which indicate that Mycoplasma pneumoniae was the cause of these outbreaks. PMID- 8246856 TI - Driver safety-belt use--Budapest, Hungary, 1993. AB - An estimated 300,000 persons die and 10-15 million persons are injured each year in traffic crashes throughout the world (1). Safety-belt use is one of the most effective means of reducing the number and severity of injuries in motor-vehicle crashes (2). In Hungary, front-seat occupants of all motor vehicles have been required to use safety belts since 1976. Since March 1993, rear-seat passengers have been required to wear safety belts in nonurban areas. Drivers in violation of the law are subject to fines and potential suspension of driving privileges. To evaluate driver compliance with the safety-belt use law, on May 10, 1993, CDC conducted an observational prevalence survey of safety-belt use in Budapest in conjunction with the U.S. Department of State and the American International School of Budapest; this survey was performed in collaboration with the Hungarian Ministry of Transport, Communication, and Water Management and the Budapest Police Department. This report presents findings of the study. PMID- 8246857 TI - Flood-related mortality--Missouri, 1993. AB - Public health surveillance documented the impact of flood-related morbidity following the floods in the midwestern United States during the summer of 1993 (1,2). Because of extensive flooding of the Missouri and Mississippi rivers and their tributaries, the Missouri Department of Health (MDH) initiated surveillance to monitor flood-related mortality. This report summarizes epidemiologic information about deaths in Missouri that resulted from riverine flooding and flash flooding during the summer and fall of 1993. PMID- 8246858 TI - Multistate outbreak of viral gastroenteritis related to consumption of oysters- Louisiana, Maryland, Mississippi and North Carolina, 1993. AB - On November 17, 1993, the state health departments of Louisiana, Maryland, and Mississippi notified CDC of several outbreaks of gastroenteritis occurring in their states since November 12. Preliminary epidemiologic investigations identified consumption of oysters as the primary risk factor for illness. On November 16, the Louisiana Department of Health and Hospitals (LDHH) had identified the Grand Pass and Cabbage Reef harvesting areas off the Louisiana coast as the source of oysters associated with outbreaks in Louisiana and Mississippi. Tagged oysters associated with outbreaks in Maryland were traced to the same oyster beds. The oysters harvested from these areas had been distributed throughout the United States. On November 18 and 19, the LDHH and CDC notified state epidemiologists of the potential for oyster-associated illness; outbreaks of oyster-associated gastroenteritis subsequently were identified in Florida and North Carolina. Collaborative investigations by state health officials, the Food and Drug Administration (FDA), and CDC were initiated to determine the magnitude and characteristics of the multistate outbreak, identify the etiologic agent, and trace the oysters. This report summarizes the preliminary findings of the ongoing investigation. PMID- 8246859 TI - HIV transmission between two adolescent brothers with hemophilia. AB - In July 1992, the National Hemophilia Foundation and CDC received a report from a hemophilia-treatment center of a 19-year-old man with hemophilia (patient 2) who recently had seroconverted for antibody to human immunodeficiency virus (HIV). This report summarizes the findings of an investigation by CDC and state and local public health officials, which determined he was infected with a strain of HIV nearly identical to that in his previously infected older brother (patient 1). PMID- 8246860 TI - Resurgence of pertussis--United States, 1993. AB - From January 3 through December 4, 1993 (weeks 1-48), 5457 pertussis cases were reported to CDC--an 82% increase over the number reported during the same period in 1992 (3004) and the highest annual number of cases reported since 1967 (Figure 1). Compared with 1992, the number of reported pertussis cases increased in 35 states, especially those in the New England, middle-Atlantic, North Central, and Mountain regions (Figure 2). During 1993, large outbreaks have occurred in Chicago and Cincinnati. This report summarizes epidemiologic characteristics of pertussis cases reported through December 4, 1993. PMID- 8246861 TI - Estimates of future global tuberculosis morbidity and mortality. AB - Tuberculosis (TB) is the leading cause of death associated with infectious diseases globally. The incidence of TB is expected to increase substantially worldwide during the next 10 years because of the interaction between the TB and human immunodeficiency virus (HIV) epidemics. This report uses TB notification data (i.e., cases reported to the ministries of health and collected by the World Health Organization [WHO]) to estimate the future global public health impact of TB and assesses the present and future contribution of HIV infection to TB. PMID- 8246862 TI - Food and Drug Administration approval of use of Haemophilus influenzae type b conjugate vaccine reconstituted with diphtheria-tetanus-pertussis vaccine for infants and children. PMID- 8246863 TI - [Ornithine decarboxylase (ODC) activity in the mucosa of human remnant stomach, and the clinical significance on tumor promotion]. AB - Ornithine Decarboxylase (ODC) is the most important and rate-determining enzyme in the biosynthetic pathway of polyamines which have received attention as potential indicators for neoplastic diseases. Here I report my results of the ODC activity in biopsy specimens of the mucosa from the remnant stomach in 45 cases followed closely within twenty years after the initial gastrectomy. I found that the mucosa at the anastomotic region showed higher ODC activity than that from any other region (p < 0.01). In all 20 cases which had undergone reconstruction Billroth II, the mucosa at the anastomotic region showed higher ODC activities than from the 20 cases which had undergone Billroth I (p < 0.05). Furthermore, the mucosa after Billroth II showed a gradually increasing mean ODC activity with time. Histologically, I found atrophy of the glands with foveolar hyperplasia more in the anastomotic region than in any other part of the remnant stomach (p < 0.001) and more so in those cases of Billroth II. These results suggest mucosal ODC activity may be a useful and effective biological indicator of the risk of carcinogenesis in the human remnant stomach. PMID- 8246864 TI - [Establishment of a transplantable rat colon cancer and a model of metastasis]. AB - A new colon cancer metastasis model was developed in inbred rat. Fischer F344 rats receiving 20mg/kg of 1,2-dimethylhydrazine per week for 20 weeks developed adenocarcinoma of the colon. The tumor has been successfully transplanted by subcutaneous inoculation for 9 generations to the present. Pathologically this transplantable tumor (TRC-1) was moderately differentiated adenocarcinoma. Single cell suspensions were obtained from TRC-1. Tumor cells (8 x 10(6)) inoculated into the portal vein of syngenic rats developed no liver metastasis. The subcutaneous tumor was excised and cultured in RPMI-1640 medium with 10% FCS. To the present, the cells were cultured for 2 years. Electron microscopic study revealed microvilli, desmosomes and intermediate filaments. The cultured cells (TRCC-1) injected into the portal vein produced liver metastases. Similarly, the cells injected into the tail vein produced lung metastases. And the cells injected into the peritoneal cavity produced peritoneal dissemination. This rat model seemed to be useful in studying the treatment for colon cancer metastasis. PMID- 8246865 TI - [The worsening of tissue damage due to the transient release during the ischemia of rat small intestine--the relation with reactive oxygen species and lipid peroxidation]. AB - Ischemia and reperfusion experiments were carried out using rat small intestine. Intestinal segment and supplying vessels was occluded and released after following procedure: In group I, 60 minute occlusion. In group II, 30 minute occlusion-10 minute release-20 minute occlusion. In group III, superoxide dismutase (SOD) and catalase (CAT) were given during the period of intermittent release in the same time schedule as in group II. Tissue damage was evaluated histopathologically and lipid peroxide (LPO) of involved segment and chemiluminescence (CL) of draining venous blood was investigated. It was concluded as follows: 1. In these experiments, the tissue damage and the increment of LPO and CL were severer in group II than in group I. Then, the reactive oxygen species (ROS) derived from neutrophils was thought to play an important role in the tissue damage. 2. At the end of intermittent release, LPO and CL increased in group II, and these increments were suppressed by SOD and CAT treatment in group III. 3. After last release, the tissue damage in group II was diminished in group III. The severe tissue damage after intermittent occlusion is thought to be caused by reperfusion injury during the intermittent release. PMID- 8246866 TI - [Prognostic value of histological findings indicating biological attitude at the invasive front of rectal cancer]. AB - Three hundred and twenty-two patients who underwent curative resection of rectal adenocarcinoma from 1970 to 1985 were studied retrospectively to evaluate the prognostic value of five histological findings at the invasive front of the lesions: 1) many small clusters of undifferentiated cancer cells ahead of the invasive front of the lesion; 2) a poorly demarcated invasive front; 3) poorly differentiated cancer cells in the invasive front; 4) intramural tumor spread; and 5) severe grade of perineural invasion. The analysis showed a significantly higher recurrence rate in the presence of each finding (p < 0.05-0.005). There was also a significant correlation between the actuarial survival rate and each finding (p < 0.01-0.001). The survival curve of Dukes' B or C tumors with at least one of those 5 findings has a significantly worse outcome than those without any (p < 0.01, respectively). Moreover, the survival curve of Dukes' B patients with one or more findings was rather worse than that of Dukes' C patients without any. In conclusion, the presence of these 5 factors at the invasive front of rectal cancer appears to indicate a vigorous biological attitude of tumor. These factors provides valuable prognostic information, independent of Dukes' stage. PMID- 8246867 TI - [The effect of oxygen supply in continuous cold perfusion of the rat liver using perfluorochemical emulsion--parenchymal and nonparenchymal cell injuries evaluated with trypan blue perfusion/fixation techniques]. AB - We investigated the effect of hypothermic aerobic perfusion of the rat liver using perfluoro-N-methyl-decahydroisoquinoline (FMIQ) on the cell viabilities of hepatic parenchyma and nonparenchyma in comparison with University of Wisconsin (UW) solution. Rat liver were perfused at 4 degrees C up to 24hr at a flow rate of 0.2 ml/g liver tissue/min with either UW or a perfusate (m-FMIQ) containing 20% FMIQ with or without oxygen saturation. After storage, livers were perfused with Krebs-Henseleit buffer containing trypan blue (200 microM). Nuclear dye uptake was measured as an indicator of cell death. Dye uptake of parenchymal cells remained less than 2% for up to 24hr, regardless of perfusate or O2 saturation. In UW perfused livers, nonparenchymal cells showed time related increases of dye positive nuclei, which were not affected by O2-saturation (39.9 +/- 4.7% and 36.5 +/- 4.2% at 24hr in O2-saturated and nonsaturated groups, respectively). In contrast, we found excellent protective effects of oxygen supply with m-EMIQ on the nonparenchymal cell viability, the dye uptake values being 16.9 +/- 5.7% and 39.4 +/- 9.1% at 24hr in O2-saturated and nonsaturated groups, respectively (p < 0.001). In conclusion, oxygen supply during hypothermic perfusion may be useful in preventing nonparenchymal cell injury of the liver, thereby leading to dramatic improvement of the hepatic microcirculation. PMID- 8246868 TI - [Unfavorable characteristics in patients with early postoperative recurrence of Graves' disease after subtotal thyroidectomy]. AB - Unfavorable characteristics of early postoperative recurrence of hyperthyroidism were studied in patients with Graves' disease after subtotal thyroidectomy. We examined 338 patients who underwent subtotal thyroidectomy during a 9-year period from 1981 to 1989. Thyroid functional status was determined biochemically and the following variables were collected and divided into two categories for each patient: sex, age at operation, indication for surgery, serum titer of antimicrosomal hemagglutination antibody, weight of resected thyroid tissue and size of remnant thyroid tissue relative to body surface area. Early postoperative recurrence was defined as relapse of the disease within one year after surgery, necessitating medication or radioiodine therapy. Logistic regression analysis disclosed that the most important characteristics were age < 20 yr, relatively large thyroid remnant and weight of resected thyroid tissue > or = 100g, the odds ratios for these variables being 20.6, 16.7 and 3.4, respectively (p < 0.05). Although candidates for surgical treatment of Graves' disease are likely to be younger and to have a large goiter, these characteristics are unfavorable, since they predict the early recurrence of hyperthyroidism. One alternative is to leave a smaller than usual thyroid remnant thyroid in these patients, even if the risk of postoperative hypothyroidism is high. PMID- 8246869 TI - [The preventive effect of additional albumin in crystalloid cardioplegic solution on myocardial reperfusion injury]. AB - The effect of additional albumin in crystalloid cardioplegic solution (CS) was investigated for infant with open heart surgery. Patients were divided into two groups; CS containing with or without 1% albumin. CS without albumin was administered for Group-I in 13 patients and CS with albumin for Group-II in 14 patients. The blood samples were collected from radial artery and coronary sinus at 5, 15 and 30 minutes after reperfusion and blood gas analysis, hemoglobin, lactate, creatine kinase-MB (CK-MB) and malondialdehyde (MDA) were measured. In percentage of CK-MB secretion, no significant differences were seen between two groups. However, in oxygen consumption, lactate and MDA secretion, significant differences between the groups were noted at five minutes after reperfusion. In conclusion, additional albumin in CS solution, especially in infant cases was effective for myocardial preservation immediately after reperfusion. PMID- 8246870 TI - [A modified composite graft: glutaraldehyde treatment of the vein and impervious covering on the composite site--an experimental and clinical study]. AB - Composite grafts have a tendency to develop intimal hyperplasia (IH) at the anastomosis between the different graft materials. The author developed a new prosthesis, modified composite graft (MCG) to control this problem. Experimental study: MCG consists of proximal 3cm of Dardik Biograft and a distal 3cm of autogenous vein (AV). Proximal segment of AV, which is anastomosed to the Biograft, was treated with 1% glutaraldehyde during operation. Then MCG was implanted in the canine infrarenal abdominal aorta. The anastomosis between Biograft and AV was wrapped with a strip of Biograft to prevent the invasion from the surrounding tissue that may also cause IH. The patency rate at 33 months was 75% (9/12). No IH was observed at Biograft-AV anastomosis. Clinical study: Thirty MCG's were implanted in patients with chronic obstructive arterial disease. None of the patients had enough autogenous vein for a complete graft. Cumulative patency rates were 67.3% at 1 year and 31.6% at 5 years. Over-all limb salvage rate was 75%. All graft failure was caused by IH at the site of proximal anastomosis between the femoral artery and Biograft or graft thrombosis probably due to low blood flow. In conclusion, IH was successfully prevented at the site of Biograft-AV anastomosis in MCG. PMID- 8246871 TI - [Leiomyosarcoma of ileocecal mesentery--a case report]. AB - We report a case of leiomyosarcoma whose origin was near the root of ileocecal mesentery and extended into the retroperitoneum. Preoperative imaging examination, including CT, MRI and angiography clearly displayed its origin and retroperitoneal extension. These results contributed to treatment planning. PMID- 8246872 TI - [A case of multiple colon cancer appeared after thymectomy for thymoma with myasthenia gravis]. AB - A 62-year-old woman was admitted to our hospital because of diarrhea and melena. Thymectomy for thymoma with myasthenia gravis was carried out on this patient four years ago. Barium enema revealed four tumor shadows in the ascending and transverse colon. Multiple colon cancer was diagnosed and extended right hemicolectomy was carried out. The postoperative course was uneventful and she has been free from any symptom. In this paper, we discussed the relationship between thymoma and the incidence of nonthymic malignancy. PMID- 8246873 TI - [Inflammatory tumor of the hepatic hilus mimicking bile duct cancer--report of a case]. AB - A 73-year-old man was admitted to our hospital because of jaundice. Ultrasonography and computed tomography showed a tumor of the hepatic hilus causing obstructive jaundice. Angiography revealed a stenosis of the right hepatic artery. Under the diagnosis of carcinoma of the hepatic hilus, extended right hepatectomy with resection of the caudate lobe was performed. A tumor showing expophytic growth, 2.1 x 1.9 x 1.5cm in size, was seen in the hepatic hilus. Histologically, the tumor was proved to be a non-neoplastic fibrous tissue simulating the inflammatory pseudotumor. PMID- 8246874 TI - [Oxygen carrying capacity of totally artificial red cell substitute--liposome embedded heme--in hemorrhagic beagle dogs]. AB - Liposome embedded heme (L/H) was produced by embedding synthesized lipid-heme in liposomes. The oxygen carrying capacity of this totally artificial oxygen carrier was evaluated in dogs. Under controlled ventilation, 30ml/kg of blood was drawn. Thirty minutes after bleeding, 30mg/kg of the L/H solution was intravenously injected in group I (n = 6) and the same volume of normal saline solution was injected in group II (n = 7). No transfusion was performed in group III (n = 7). In group I, mixed venous O2 tension (PvO2), O2 transport (TO2) and O2 consumption (VO2) increased significantly compared with those observed before injection of the L/H (p < 0.05). PvO2 kept higher level compared with group II until 90 minutes after injection of the L/H (p < 0.05). Following the L/H injection, TO2 by the dogs' Hb was 187ml/min and that by the L/H was 43ml/min. VO2 from the dogs' Hb was 51ml/min and that from the L/H was 16ml/min. Thus TO2 by the L/H provided 19% of the total TO2 and VO2 from the L/H provided 24% of the total VO2. Thus, this exchange transfusion experiment show that the L/H has ability as an oxygen carrier to bind and to transport oxygen to the tissues. PMID- 8246875 TI - [Utility of measurement of tumor markers for preoperative staging of gastric cancer]. AB - In 120 operated cases of gastric cancer, nine tumor makers (serum carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), carbohydrate antigen 19 9 (CA19-9), carbohydrate antigen 125 (CA125), tissue polypeptide antigen (TPA), serum beta 2-microglobulin (s-BMG), urine beta 2-microglobulin (mu-BMG), serum immunosuppressive acidic protein (IAP) and sialic acid (SA)) were determined before operation, to explore distribution patterns, positive rates, and relationships among these markers. And, we explored tumor markers which would be useful in preoperative assessment of the cancer stage and feasibility of radical treatment. The following results were obtained. 1. The distribution of all tumor markers was close to normal logarithmic distribution. 2. The positive rate was high in the order of IAP > TPA > CEA > CA19-9 >s-BMG > SA > u-BMG > CA125 > AFP. 3. Strong correlations were observed between IAP and SA, between CA125 and TPA. 4. The optimum combination of markers for diagnosis of the cancer stage was IAP + CA19-9 + CA125 and the optimum combination for assessment of the feasibility of radical treatment was IAP + CA125 + TPA. PMID- 8246876 TI - [Thymidine kinase and thymidylate synthetase activities in human gastric cancer]. AB - Thymidine kinase (TK) and thymidylate synthetase (TS) are known to catalyse the phosphorylation of thymidine for the salvage synthesis of dTMP and the methylation of dUMP for the de novo synthesis of dTMP, respectively. High TK and TS activities and the existence of TK isozymes have been observed in rapidly proliferating tissues. In this work I measured TK, TK-isozyme and TS activities, and DNA and RNA in human gastric cancer tissue. Tissue containing gastric cancer, (well differentiated adenocarcinoma (well: n = 29), poorly differentiated adenocarcinoma (por: n = 28)), and noninvolved paired normal gastric mucosa were obtained from fresh surgical specimens. Total TK and peak A TK activity of gastric cancer increased to 184% and 299% of activity of normal gastric mucosa. TK activity of "well" was higher than that of "por". On the other hand TS activity of gastric cancer increased to 122% of activity of normal gastric mucosa. TS activity of "por" was higher than that of "well". DNA and RNA content of gastric cancer increased to 294% and 228% than that of normal gastric mucosa. But difference wasn't observed between "well" and "por". These characteristics of gastric cancer suggest its susceptibility to control by inhibition of TK and TS. PMID- 8246877 TI - Cloning of Saccharomyces cerevisiae STE5 as a suppressor of a Ste20 protein kinase mutant: structural and functional similarity of Ste5 to Far1. AB - The beta and gamma subunits of the mating response G-protein in the yeast Saccharomyces cerevisiae have been shown to transmit the mating pheromone signal to downstream components of the pheromone response pathway. A protein kinase homologue encoded by the STE20 gene has recently been identified as a potential G beta gamma target. We have searched multicopy plasmid genomic DNA libraries for high gene dosage suppressors of the signal transduction defect of ste20 mutant cells. This screen identified the STE5 gene encoding an essential component of the pheromone signal transduction pathway. We provide genetic evidence for a functional interrelationship between the STE5 gene product and the Ste20 protein kinase. We have sequenced the STE5 gene, which encodes a predicted protein of 917 amino acids and is specifically transcribed in haploid cells. Transcription is slightly induced by treatment of cells with pheromone. Ste5 has homology with Far1, a yeast protein required for efficient mating and the pheromone-inducible inhibition of a G1 cyclin, Cln2. A STE5 multicopy plasmid is able to suppress the signal transduction defect of far1 null mutant cells suggesting that Ste5, at elevated levels, is able functionally to replace Far1. The genetically predicted point of function of Ste5 within the pheromone signalling pathway suggests that Ste5 is involved in the regulation of a G beta gamma-activated protein kinase cascade which links a G-protein coupled receptor to yeast homologues of mitogen activated protein kinases. PMID- 8246878 TI - The Streptomyces lividans 66 chromosome contains a 1 MB deletogenic region flanked by two amplifiable regions. AB - Genetic instability in Streptomyces species often involves large deletions sometimes accompanied by DNA amplification. Two such systems in Streptomyces lividans 66 involve the production of mutants sensitive to chloramphenicol and the production of mutants resistant to the galactose analogue 2-deoxygalactose, respectively. Overlapping cosmids were isolated that span the ca. 1 Mb region between the two amplifiable regions. The structure of the region was confirmed by restriction mapping using the rarely cutting enzymes AseI, BfrI and DraI and pulsed-field gel electrophoresis. The region contains a non-clonable gap flanked by inverted repeats; the structure is consistent with the presence of a physical gap, i.e. a linear chromosome. PMID- 8246879 TI - Expression of beta 1 tubulin (beta Tub56D) in Drosophila testis stem cells is regulated by a short upstream sequence while intron elements guide expression in somatic cells. AB - Stem cell differentiation to mature spermatozoa is a morphogenetic process that is highly dependent on microtubular arrays. In the early, mitotically active stages of spermatogenesis, only the beta 1 tubulin isotype is expressed. Analysis of transgenic flies containing beta 1-lacZ gene fusions revealed that this expression is regulated by sequences located between positions -45 and -191 upstream of the transcription initiation site. Furthermore, beta 1 tubulin is a major component of cyst cells. Expression in these cells is driven by enhancer elements located in the beta 1 tubulin gene intron. These enhancer elements also guide expression in combination with the hsp70 basal promoter. In addition, redundant enhancer elements in the intron drive expression in the testis wall. Our data show that within a single tissue, the male gonad, expression of the beta 1 tubulin gene is under cell-type-specific control mediated by independent cis acting elements. Therefore in the germ line, control of beta 1 tubulin expression is strictly governed by promoter-proximal elements, while for the somatic parts of the testis, enhancer elements confer less stringent expression control. PMID- 8246880 TI - A variable DNA region of Entamoeba histolytica is expressed in several transcripts which differ in genetically related clones. AB - A highly variable DNA region (EhVR1), isolated from Entamoeba histolytica clone A, strain HM1:IMSS, is transcribed into several transcripts, which differ in genetically related clones. EhVR1 (3.5 kb) is composed of two contiguous fragments; one of these 1.9 kb long, at the 3' end, identified similar transcripts in clones A, L6 and C2 (all derived from strain HM1:IMMS), the other of 1.6 kb, at the 5' end, detected 0.5, 0.6 and 0.7 kb transcripts only in clone A. Variability of the 1.6 kb fragment was found even within the same clone maintained under different conditions. EhVR1 was localized to 1.3 and 1.4 Mb linear chromosomes and also found in circular molecules. The sequence of the 1.6 kb fragment revealed the presence of a large number of different repeats, including inverted and palindromic repeats. A p145 sequence, previously detected in episomal DNA of the amoeba, was found at the 5' end of EhVR1. The presence of EhVR1 in linear and circular molecules, its high number of repeats, and its variability in genetically related clones suggest the existence of DNA regions that undergo dynamic non-reciprocal recombination between circular episomes and linear chromosomes, and may thus contribute to variability in the trophozoite genome. PMID- 8246881 TI - A dominant interfering mutation in RAS1 of Saccharomyces cerevisiae. AB - A mutant allele of RAS1 that dominantly interferes with the wild-type Ras function in the yeast Saccharomyces cerevisiae was discovered during screening of mutants that suppress an ira2 disruption mutation. A single amino acid substitution, serine for glycine at position 22, was found to cause the mutant phenotype. The inhibitory effect of the RAS1Ser22 gene could be overcome either by overexpression of CDC25 or by the ira2 disruption mutation. These results suggest that the RAS1Ser22 gene product interferes with the normal interaction of Ras with Cdc25 by forming a dead-end complex between Ras1Ser22 and Cdc25 proteins. PMID- 8246882 TI - Integration of repeated sequences (pBR322) in the Bacillus subtilis 168 chromosome without affecting the genome structure. AB - The Escherichia coli plasmid pBR322 sequence (4363 bp) was integrated at the met, pro, or leuB locus of the Bacillus subtilis chromosome without duplication of the flanking chromosomal regions. The integrated pBR322 was stably maintained as part of the chromosome regardless of its orientation or location. It was found that a DNA segment as large as 17 kb cloned in pBR322 can be readily transferred to the B. subtilis chromosome by transformation. It was demonstrated that a second pBR322 sequence could be effectively introduced at different regions of the chromosome by sequential transformation using chromosomal DNA isolated from a strain that had already acquired a pBR322 sequence at a different locus. Similarly, a third pBR322 sequence could be introduced. By this method, two or three pBR322 sequences can be incorporated at unlinked loci without affecting the overall structure of the B. subtilis genome. PMID- 8246883 TI - The amiloride resistance gene, car1, of Schizosaccharomyces pombe. AB - Amiloride, an inhibitor of various sodium transporters, is toxic to Schizosaccharomyces pombe at low concentration in minimal but not in rich media. Amiloride-resistant mutants were isolated and shown to represent a new locus (car1 for changed amiloride resistance) on chromosome I. The car1 gene was cloned and sequenced. Sequence analysis revealed an open reading frame of 526 amino acids with a predicted molecular weight of 58,545 Da. It has 52% hydrophobic residues and belongs to the class of 12-transmembrane-domain transport proteins. Gene disruption of car1 results in increased amiloride resistance. car1 has sequence similarity to proteins from Candida associated with resistance to benomyl, methotrexate and cycloheximide. No single physiologically identifiable component of sodium transport appeared to be lost. We propose that car1 serves an uptake function, perhaps as a symport with an unknown substrate and this carrier may transport amiloride into the cell. Further, we suggest that amiloride toxicity at low concentrations is not due to its effect on sodium transport but, rather, depends on intracellular interference with an unknown biosynthetic pathway. PMID- 8246884 TI - The HYP2 gene of Saccharomyces cerevisiae is essential for aerobic growth: characterization of different isoforms of the hypusine-containing protein Hyp2p and analysis of gene disruption mutants. AB - In Saccharomyces cerevisiae, hypusine-containing proteins are encoded by two closely related genes, HYP1 and HYP2, which are regulated reciprocally by oxygen and heme. We have purified the aerobically expressed hypusine-containing proteins from yeast. The three proteins detected (two isoforms, which differ in their pI values, and a degradation product thereof, lacking the N-terminal 10 amino acid residues) are all encoded by HYP2. The N-terminus of both isoforms is formed by acetylation of a serine residue after cleavage of the first methionine. Cells mutant for hyp2 are unable to grow aerobically. However, under anaerobic conditions these mutants display no obvious phenotype, presumably because the strictly anaerobically expressed HYP1 gene product (Hyp1p) is present. This implies that Hyp1p and Hyp2p fulfill very similar functions. In fact, Hyp1p can substitute for Hyp2p under aerobic conditions, when expressed under the control of the GAL1 promoter in hyp2 mutant cells. PMID- 8246885 TI - Characterization of an aspartic proteinase of Mucor pusillus expressed in Aspergillus oryzae. AB - The aspartic proteinase (MPP) gene from the zygomycete fungus Mucor pusillus was introduced into an ascomycete fungus, Aspergillus oryzae, by protoplast transformation using the nitrate reductase (niaD) gene as the selective marker. Southern blot analysis indicated that the MPP gene was integrated into the resident niaD locus at a copy number of 1-2. MPP secreted by the recombinant A. oryzae was correctly processed but was more highly glycosylated than that produced in the original M. pusillus strain. Treatment with endo-beta-N acetylglucosaminidase H and analysis of the carbohydrate composition of the secreted MPP revealed that the extra glycosylation of the MPP secreted by the recombinant A. oryzae was due to altered processing of mannose residues. The extra glycosylation of MPP affected its enzyme properties including its milk clotting and proteolytic activities. PMID- 8246886 TI - Functional expression of the transcriptional activator Opaque-2 of Zea mays in transformed yeast. AB - The aim of this research was to determine whether the structural homology between the O2 gene, a maize transcriptional activator, and the GCN4 gene, a yeast transcriptional factor, is reflected at the level of function. The O2 cDNA was cloned in the yeast expression vector pEMBLyex4 under the control of a hybrid inducible promoter, and used to transform the yeast Saccharomyces cerevisiae. Transformed yeast cells produced O2 mRNA and a polypeptide immunoreactive with anti-O2 antibodies during growth in galactose. The heterologous protein was correctly translocated into the yeast nuclei, as demonstrated by immunofluorescence, indicating that the nuclear targeting sequences of maize are recognized by yeast cells. Further experiments demonstrated the ability of O2 to rescue a gcn4 mutant grown in the presence of aminotriazole, an inhibitor of the HIS3 gene product, suggesting that O2 activates the HIS3 gene, gene normally under control of GCN4. It was shown that the O2 protein is able to trans-activate the HIS4 promoter in yeast cells and binds to it in vitro. The sequence protected by O2, TGACTC, is also the binding site for GCN4. Finally, the expression of O2 protein in yeast did not produce alterations during batch growth at 30 degrees C, while transformants expressing O2 protein showed a conditionally lethal phenotype when grown in galactose at 36 degrees C; this phenotype mimics the behaviour of gcd mutants. The results support the idea that basic mechanisms of transcription control have been highly conserved in eukaryotes. PMID- 8246887 TI - An impaired RNA polymerase II activity in Saccharomyces cerevisiae causes cell cycle inhibition at START. AB - Saccharomyces cerevisiae cells harboring the temperature-sensitive mutation rpo21 4, in the gene encoding the largest subunit of RNA polymerase II, were shown to be partially impaired for cell-cycle progress at a permissive temperature, and to become permanently blocked at the cell-cycle regulatory step, START, at a restrictive temperature. The rpo21-4 mutation was lethal in combination with cdc28 mutations in the p34 protein kinase gene required for START. Transcripts of the CLN1 and CLN2 genes, encoding G1-cyclin proteins that, along with p34, are necessary for START, were decreased in abundance by the rpo21-4 mutation at a restrictive temperature. Increased G1-cyclin production, by expression of the CLN1 or CLN2 genes from a heterologous GAL promoter, overcame the rpo21-4 mediated START inhibition, but such mutant cells nevertheless remained unable to proliferate at a restrictive temperature. These findings reveal that START can be particularly sensitive to an impaired RNA polymerase II function, presumably through effects on G1-cyclin expression. PMID- 8246888 TI - Molecular analysis of the major cellulase (CelV) of Erwinia carotovora: evidence for an evolutionary "mix-and-match" of enzyme domains. AB - The structural gene for the major cellulase of Erwinia carotovora subspecies carotovora (Ecc) was isolated and expressed in Escherichia coli. Sequencing of the gene (celV) revealed a typical signal sequence and two functional domains in the enzyme; a catalytic domain linked by a short proline/threonine-rich linker to a cellulose-binding domain (CBD). The deduced amino acid sequence of the catalytic domain showed homology with cellulases of Family A, including enzymes from Bacillus spp. and Erwinia chrysanthemi CelZ, whereas the CBD showed homology with cellulases from several diverse families, supporting a "mix-and-match" hypothesis for evolution of this domain. Analysis of the substrate specificity of CelV showed it to be an endoglucanase with some exoglucanase activity. The pH optimum is about 7.0 and the temperature optimum about 42 degrees C. CelV is secreted by Ecc and by the taxonomically related Erwinia carotovora subspecies atroseptica (Eca) but not by E. coli. Overproduction of the enzyme from multicopy plasmids in Ecc appears to overload the secretory mechanism. PMID- 8246889 TI - Cloning of the chitin synthase 3 gene from Candida albicans and its expression during yeast-hyphal transition. AB - The chitin synthase 3 gene (CACHS3) has been cloned from Candida albicans. The yeast CAL1 gene encoding the chitin synthase 3 of Saccharomyces cerevisiae was used as a probe for the isolation of the gene from C. albicans. The CAL1 homolog was identified in Southern blots of C. albicans genomic DNA and cloned from a C. albicans genomic DNA library. The nucleotide sequences of two partial clones were determined and combined giving a total length of 4610 bp. A continuous open reading frame of 3525 bp encoding a predicted protein of 1175 amino acids and molecular mass of 131 850 daltons was identified. A comparison of the deduced amino acid sequences of CAL1 and the Candida chitin synthase 3 protein showed 59.3% identity. Southern blot analysis indicates that the CACHS3 gene is present in a single copy in the genome and maps to chromosome I. Northern blot analysis shows that expression of chitin synthase 3 gene is dramatically increased during the transition from the yeast to the hyphal form of C. albicans. This change in transcription level strongly suggests that CACHS3 may play a role in Candida morphogenesis. PMID- 8246890 TI - Stable expression of a single-copy rolA gene in transgenic Arabidopsis thaliana plants allows an exhaustive mutagenic analysis of the transgene-associated phenotype. AB - Several publications have documented the instability of transgene expression in plants. Previous genetic approaches to the study of transgene-associated phenotypes in plants were limited by this phenomenon. Here we show that a transgene can be expressed in plants with sufficient stability to allow an exhaustive mutagenic analysis of the resulting phenotype. We have expressed the morphogenic rolA gene from the TL-DNA of Agrobacterium rhizogenes Ri plasmid in transgenic Arabidopsis thaliana plants. The resulting pleiotropic RolA phenotype allows a visual screen for reversion to detect germinal as well as somatic instability of transgene expression. However no spontaneous reversions of the Ro 1A phenotype were observed in 65,000 progeny of two independent transgenic A. thaliana lines, each carrying a single homozygous rolA locus. In contrast, 12 revertants of the RolA phenotype were isolated from 360,000 ethyl methane sulphonate (EMS)-mutagenized M2 progeny. All revertants were shown genetically to carry stable recessive mutations in the rolA locus, thus establishing a series of loss-of-function alleles. Molecular characterization revealed that the loss-of function alleles were structurally intact and expressed in all rolA mutants. A wildtype rolA locus and two loss-of-function alleles were reisolated and sequenced; base pair substitutions were found in each loss-of-function allele leading to single amino acid substitutions in the rolA open reading frame. Therefore no instability of expression of the rolA locus was detected in any of the 425,000 individuals studied in this analysis. Furthermore even under conditions of saturation mutagenesis, no extragenic suppressor locus was detected. PMID- 8246891 TI - Identification and analysis of the Rhizobium meliloti exoAMONP genes involved in exopolysaccharide biosynthesis and mapping of promoters located on the exoHKLAMONP fragment. AB - Sequence analysis of a 7.494 kb DNA fragment from megaplasmid 2 of Rhizobium meliloti 2011 involved in exopolysaccharide I (EPS I) biosynthesis revealed the presence of five exo genes designated exoA, exoM, exoN, exoO, and exoP. ExoN was found to show strong homology to a UDP-glucose pyrophosphorylase from Acetobacter xylinum, whereas ExoO displayed weak homologies to the NodC proteins from R. meliloti and R. loti. Surprisingly, different mutations in exoP resulted in divergent phenotypes. One exoP mutant was able to establish an effective symbiosis with alfalfa, although no EPS I polymer could be detected. In contrast, other exoP mutations prevented the formation of an effective symbiosis. The transcriptional organization of the exoA-exoP gene region has been analysed in conjunction with the exoH, exoK and exoL genes. Using exo-lacZ transcription fusions in association with plasmid integration mutagenesis a strong promoter was identified upstream of exoH, which is able to direct transcription of the whole exoHKLAMONP gene cluster. A much weaker promoter upstream of exoL was found to be involved in the transcription of the exoLAMONP genes. In addition, weak promoters were identified upstream of exoK, exoA, exoN and exoP. PMID- 8246892 TI - Transactivation of Ds elements in plants of lettuce (Lactuca sativa). AB - The maize transposable element, Activator (Ac), is being used to develop a transposon mutagenesis system in lettuce, Lactuca sativa. In this paper, we describe somatic and germinal transactivation of Ds by chimeric transposase genes in whole plants. Constructs containing either the Ds element or the Ac transposase open reading frame (ORF) were introduced into lettue. The Ds element was located between either the 35S or the Nos promoter and a chimeric spectinomycin resistance gene (which included a transit peptide), preventing expression of spectinomycin resistance. The genomic coding region of the Ac transposase was expressed from the 35S promoter. Crosses were made between 104 independent R1 plants containing Ds and three independent R1 plants expressing transposase. The excision of Ds in F1 progenies was monitored using a phenotypic assay on spectinomycin-containing medium. Green sectors in one-third of the F1 families indicated transactivation of Ds by the transposase at different developmental stages and at different frequencies in lettuce plants. Excision was confirmed using PCR and by Southern analysis. The lack of green sectors in the majority of F1 families suggest that the majority of T-DNA insertion sites are not conducive to excision. In subsequent experiments, the F1 plants containing both Ds and the transposase were grown to maturity and the F2 seeds screened on medium containing spectinomycin. Somatic excision was again observed in several F2 progeny; however, evidence for germinal excision was observed in only one F2 family. PMID- 8246894 TI - Deletion of the mating-type sequences in Podospora anserina abolishes mating without affecting vegetative functions and sexual differentiation. AB - The mating-type locus of Podospora anserina controls fusion of sexual cells as well as subsequent stages of development of the fruiting bodies. The two alleles at the locus are defined by specific DNA regions comprising 3.8 kb for mat+ and 4.7 kb for mat-, which have identical flanking sequences. Here we present the characterization of several mutants that have lost mat(+)-specific sequences. One mutant was obtained fortuitously and the other two were constructed by gene replacement. The mutants are deficient in mating with strains of either mat genotype but are still able to differentiate sexual reproductive structures. The loss of the mating type does not lead to any discernible phenotype during vegetative growth: in particular it does not change the life span of the strain. The mutants can recover mating ability if they are transformed with DNA containing the complete mat+ or mat- information. The transformants behave in crosses as do the reference mat+ or mat- strains, thus indicating that the transgenic mat+ and mat- are fully functional even when they have integrated at ectopic sites. PMID- 8246893 TI - Identification, molecular cloning and characterization of the gene encoding the chi subunit of DNA polymerase III holoenzyme of Escherichia coli. AB - We have identified a previously reported open reading frame (ORF13) that maps between pepA and valS at 96.6 centisomes of the Escherichia coli genome as the structural gene for the chi subunit of DNA polymerase III holoenzyme. This conclusion is supported by a perfect match of the amino-terminal 24 residues of chi with the DNA sequence of ORF13 and a demonstration that ORF13 directs expression of a protein that co-migrates with authentic chi on SDS-polyacrylamide gels. ORF13, designated holC, was isolated from the E. coli chromosome and inserted into a tac promoter-based expression plasmid to direct production of the chi subunit to 5-7% of the total soluble protein. The 3' end of holC was sequenced to resolve discrepancies between two published versions. PMID- 8246895 TI - Genes expressed during sexual differentiation of Chlamydomonas reinhardtii. AB - Four genes specifically expressed during gametogenesis of Chlamydomonas reinhardtii have been cloned and their expression patterns analyzed. mRNAs encoded by these gamete-specific genes (gas) were absent or present only at very low levels in vegetative cells and mature zygotes. In young zygotes 2 h after gamete fusion, the mRNAs of three gas genes still persisted. The gas mRNAs accumulated during gametic differentiation. The temporal patterns of accumulation of individual mRNAs differed; some started to increase early during gametogenesis, others accumulated in the late phase. The accumulation of one of the late mRNAs (gas28) was strictly light-dependent. To illustrate the utility of the genes cloned in the analysis of sexual differentiation in Chlamydomonas reinhardtii we show that in a gametogenesis-defective mutant, the expression of late genes is prevented while that of early genes is normal. PMID- 8246896 TI - Cloning of the isocitrate lyase gene (ICL1) from Yarrowia lipolytica and characterization of the deduced protein. AB - The ICL1 gene encoding isocitrate lyase was cloned from the dimorphic fungus Yarrowia lipolytica by complementation of a mutation (acuA3) in the structural gene of isocitrate lyase of Escherichia coli. The open reading frame of ICL1 is 1668 bp long and contains no introns in contrast to currently sequenced genes from other filamentous fungi. The ICL1 gene encodes a deduced protein of 555 amino acids with a molecular weight of 62 kDa, which fits the observed size of the purified monomer of isocitrate lyase from Y. lipolytica. Comparison of the protein sequence with those of known pro- and eukaryotic isocitrate lyases revealed a high degree of homology among these enzymes. The isocitrate lyase of Y. lipolytica is more similar to those from Candida tropicalis and filamentous fungi than to Saccharomyces cerevisiae. This enzyme of Y. lipolytica has the putative glyoxysomal targeting signal S-K-L at the carboxy-terminus. It contains a partial repeat which is typical for eukaryotic isocitrate lyases but which is absent from the E. coli enzyme. Surprisingly, deletion of the ICL1 gene from the genome not only inhibits the utilization of acetate, ethanol, and fatty acids, but also reduces the growth rate on glucose. PMID- 8246897 TI - Suppression of phaseolin and lectin in seeds of common bean, Phaseolus vulgaris L.: increased accumulation of 54 kDa polypeptides is not associated with higher seed methionine concentrations. AB - Suppression of phaseolin and lectin accumulation in common bean resulted in higher concentrations of bean seed polypeptides with apparent molecular weights of 54 kDa and from 70 to 84 kDa on SDS-polyacrylamide gel electrophoresis. Polypeptides of 54 and 56 kDa segregated as products of different alleles. Genes for the 54/56 kDa bands and phaseolin were estimated to be 26.2 +/- 3.7 map units apart. The 54 kDa band phenotype manifested by SDS-PAGE consisted of from one to three polypeptides of 54 kDa MW on 2D gels, and the 56 kDa phenotype consisted of one polypeptide of 56 kDa plus two minor polypeptides of 54-54.5 kDa molecular weight. The pKI of these polypeptides was approximately 5.25. The methionine content of the 54 kDa polypeptides of the cultivar Great Northern Star was 1.6 +/ 0.1 g/100 g protein, which was not statistically different from the value (1.5 +/- 0.1%) obtained for phaseolin isolated by the same procedure. F2 seeds deficient for phaseolin and lectin contained as much total N per g as wild-type seeds and were not shrunken, but contained 50% more free amino acids. F2 seeds from two of the three populations contained from 8 to 13% less methionine per mg total N. PMID- 8246898 TI - Sequence complexity of the S receptor kinase gene family in Brassica. AB - A genomic library from an S29/S29 self-incompatible genotype of Brassica oleracea was screened with a probe carrying part of the catalytic domain of a Brassica S receptor kinase (SRK)-like gene. Six positive phage clones with varying hybridisation intensities (K1 to K6) were purified and characterised. A 650-700 bp region corresponding to the probe was excised from each clone and sequenced. DNA and predicted protein sequence comparisons based on a multiple alignment identified K5 as a pseudogene, whereas the others could encode functional proteins. K3 was found to have lost an intron from its genomic sequence. The six genes display different degrees of sequence similarity and form two distinct clusters in a dendrogram. The 98% similarity between K4 and K6, which extends across intron sequences, suggests that these might be very recently diverged alleles or daughters of a duplication. In addition, K2 showed a comparably high similarity to the probe. Clones K1, K3 and K5 cross-hybridised with an SLG29 cDNA probe, indicating the presence of upstream receptor domains homologous to the Brassica SLG gene. This suggests that the previously reported S sequence complexity may be ascribed to a large receptor kinase gene family. PMID- 8246899 TI - The vacuolar H(+)-ATPase of Saccharomyces cerevisiae is required for efficient copper detoxification, mitochondrial function, and iron metabolism. AB - Mutations in the GEF2 gene of the yeast Saccharomyces cerevisiae have pleiotropic effects. The gef2 mutants display a petite phenotype. These cells grow slowly on several different carbon sources utilized exclusively or primarily by respiration. This phenotype is suppressed by adding large amounts of iron to the growth medium. A defect in mitochondrial function may be the cause of the petite phenotype: the rate of oxygen consumption by intact gef2 cells and by mitochondrial fractions isolated from gef2 mutants was reduced 60%-75% relative to wild type. Cytochrome levels were unaffected in gef2 mutants, indicating that heme accumulation is not significantly altered in these strains. The gef2 mutants were also more sensitive than wild type to growth inhibition by several divalent cations including Cu. We found that the cup5 mutation, causing Cu sensitivity, is allelic to gef2 mutations. The GEF2 gene was isolated, sequenced, and found to be identical to VMA3, the gene encoding the vacuolar H(+)-ATPase proteolipid subunit. These genetic and biochemical analyses demonstrate that the vacuolar H(+)-ATPase plays a previously unknown role in Cu detoxification, mitochondrial function, and iron metabolism. PMID- 8246900 TI - In vivo species specificity of DNA polymerase alpha. AB - The DNA polymerase alpha enzymes from human, and budding (Saccharomyces cerevisiae) and fission yeast (Schizosaccharomyces pombe) are homologous proteins involved in initiation and replication of chromosomal DNA. Sequence comparison of human DNA polymerase alpha with that of S. cerevisiae and S. pombe shows overall levels of amino acid sequence identity of 32% and 34%, respectively. We report here that, despite the sequence conservation among these three enzymes, functionally active human DNA polymerase alpha fails to rescue several different conditional lethal alleles of the budding yeast POL1 gene at nonpermissive temperature. Furthermore, human DNA polymerase alpha cannot complement a null allele of budding yeast POL1 either in germinating spores or in vegetatively growing cells. In fission yeast, functionally active human DNA polymerase alpha is also unable to complement the disrupted pol alpha::ura4+ allele in germinating spores. Thus, in vivo, DNA polymerase alpha has stringent species specificity for initiation and replication of chromosomal DNA. PMID- 8246901 TI - The role of tyrosine-114 in the enzymatic activity of the Shiga-like toxin I A chain. AB - Shiga-like toxin I (SLT-I), the potent cytotoxin produced by certain pathogenic strains of Escherichia coli, is a member of a burgeoning family of ribosome-in activating proteins (RIPs), which share common structural and mechanistic features. The prototype of the group is the plant toxin ricin. Recently we proposed a structural model for the Slt-IA active site, based in part on the known geometry of the enzymatic subunit of the ricin toxin. The model places three aromatic residues within the putative Slt-IA active site cleft: tyrosine 77, tyrosine 114, and tryptophan 203. Here we present biochemical and biophysical data regarding, the phenotypes of conservative point mutants of Slt-IA in which tyrosine 114 is altered. We used oligonucleotide-directed mutagenesis to replace tyrosine 114 with either phenylalanine (Y114F) or serine (Y114S). Periplasmic extracts of E. coli containing wild-type or mutant Slt-IA were tested for their ability to inhibit protein synthesis in vitro. Relative to wild-type, the activity of mutant Y114F was attenuated about 30-fold, while the mutant Y114S was attenuated about 500 to 1000-fold. In order to address the possibility that differential activation of the mutants rather than local effects at the active site might account for their diminished activity, we engineered the same mutations into a truncated slt-IA cassette that directs expression of a product corresponding to the activated A1 form of Slt-IA (wild-type-delta). The same general relationships held: relative to wild type-delta, Y114F-delta was attenuated about 7-fold, and Y114S-delta about 300-fold.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8246902 TI - Expression of A mating type genes of Coprinus cinereus in a heterologous basidiomycete host. AB - The A mating factor of Coprinus cinereus determines compatibility in mating by regulating part of a developmental sequence that leads to dikaryon formation. The A genes that trigger development encode two different classes of homeodomain proteins, and for a successful mating, a protein of one class, HD1, must interact with a protein of the other class, HD2. In this report we show that C. cinereus A genes that encode HD2 proteins, a2-1 and b2-1, can elicit A-regulated development in the heterologous host C. bilanatus. Transformation rates were very low, suggesting that the genes were poorly transcribed. The fact that the HD2 genes are functionally expressed implies successful heteromultimeric association of putative DNA-binding proteins coded by the two Coprinus species. This interaction was sufficient to satisfy the need for different A factors in the formation of a fertile C. bilanatus dikaryon, but fertile dikaryons were more readily produced in matings with the a2-1 gene transformants. The C. cinereus A genes, b1-1 and d1 1, which encode HD1 proteins, were either not expressed or their proteins were non-functional in C. bilanatus. These experiments raise some interesting questions regarding HD1-HD2 protein interactions. PMID- 8246903 TI - The sugar beet mitochondrial genome contains an ORF sharing sequence homology with the gene for the 30 kDa subunit of bovine mitochondrial complex I. AB - From a sugar beet mitochondrial DNA library, we have isolated an open reading frame (ORF192) showing extensive homology to the gene for the 30 kDa subunit of the bovine mitochondrial complex I (NADH: ubiquinone reductase). The ORF192 was found to be actively transcribed to give an RNA of approximately 1.0 kb. We have designated this gene nad9. Transcripts from the nad9 locus are edited by five C to U transitions, increasing similarity with the amino acid sequence of the corresponding bovine and Neurospora crassa polypeptides. Southern blot hybridization also indicates that nad9 is present in the mitochondrial genomes of a variety of higher plant species. PMID- 8246904 TI - Structure-activity relationships for cannabinoid receptor-binding and analgesic activity: studies of bicyclic cannabinoid analogs. AB - Cannabimimetic compounds, such as delta 9-tetrahydrocannabinol (delta 9-THC), evoke analgesia in addition to other behavioral responses in humans and animals. The cannabinoid receptor mediating this response has been characterized by its ability to bind the cannabinoid agonist [3H]CP-55,940 and to inhibit adenylyl cyclase via Gi. An investigation of structural requirements for antinociceptive activity of cannabinoid structures led to the development of a simple bicyclic cannabinoid agonist, CP-47,497, that possessed a spectrum of cannabinoid activities in animals that resembled that of delta 9-THC. The present investigation examines several series of CP-47,497 analogs for their binding affinity at the cannabinoid receptor and their ability to evoke analgesia in rodents. Analogs substituted at the C-3 alkyl side chain exhibited maximal affinity for the cannabinoid receptor with side chains of seven or eight carbons in length. Analgesic potency paralleled the receptor-binding affinity. The cyclohexyl ring was optimized as a six- or seven-membered ring structure for binding as well as analgesic activity. Cyclohexyl alkyl side chain extensions of up to four carbons in length had little influence on the affinity for the receptor or analgesic activity. Hydroxyalkyl side chains exhibited optimal binding affinity and antinociceptive activity at three or four carbon atoms in length; however, polar groups closer to the ring diminished binding to the receptor. The importance of the phenolic and cyclohexyl hydroxyl groups for binding affinity was demonstrated. In general, analgesic activity correlated well with the affinity of these analogs for the cannabinoid receptor. Exceptions could be explained by metabolic transformations likely to occur in vivo. PMID- 8246905 TI - Modulation of rat thymocyte proliferative response through the inhibition of different cyclic nucleotide phosphodiesterase isoforms by means of selective inhibitors and cGMP-elevating agents. AB - We have investigated the role played by cyclic nucleotide phosphodiesterases (EC 3.1.4.17) in the control of T-lymphocyte response to mitogenic agents by their ability to influence the cellular level of cAMP. The importance of this messenger as a negative regulator in this cell type is well established. Multiple isoenzymes of phosphodiesterase were fractionated from the cytosol of rat thymic lymphocytes by high performance liquid chromatography on an anion exchange column. In addition to the type II, III, IV isoforms that we have already described [Valette et al., Biochem. Biophys. Res. Commun. 169:864-872 (1990)], a phosphodiesterase fraction sharing several of the characteristics of type V, cGMP binding phosphodiesterase, was detected. Non-isoform-selective inhibitors of phosphodiesterase such as dipyridamole, papaverine, and methyl-isobutylxanthine were able to totally prevent the proliferative response of thymocytes to stimulation by the mitogenic lectin concanavalin A. In contrast, the selective inhibitor of type IV phosphodiesterases rolipram induced a rather moderate inhibition of proliferation, not exceeding 60%; and the selective inhibitors of type III and type V phosphodiesterases, milrinone and M&B 22,948, respectively, displayed only marginal inhibitory effects. The association of the type III and IV phosphodiesterase inhibitors produced synergistic inhibition of proliferation, which could then be almost totally suppressed. These inhibitory effects on cell multiplication were reflected at the level of the cell cAMP content; only rolipram was able to induce a significant (approximately 50%) increase in cAMP, and this increase was potentiated by the presence of milrinone, reaching almost 100%. The type V phosphodiesterase selective inhibitor M&B 22,948 displayed similar properties to those of milrinone, which suggests that it indirectly inhibited the type III, cGMP-inhibitable isoenzyme, by inducing a cGMP rise. This hypothesis was supported by evidence of a significant raising effect of M&B 22,948 on cGMP level, and by the ability of a cGMP-elevating agent, sodium nitroprusside, to mimic the synergistic effects of milrinone associated with rolipram. Furthermore, 8-bromo-cGMP, a potent activator of cGMP-dependent protein kinase, which showed only weak inhibitory effects on thymic type III phosphodiesterase, failed to alter the effects of rolipram on the cell proliferation. These results allow us to delineate a role for types III, IV, and V phosphodiesterase in the control of cAMP level during the proliferative response of thymic lymphocytes. They also suggest that endogenously formed cGMP might participate in the regulation of cAMP level in the cells by means of the inhibition of the type III phosphodiesterase.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246906 TI - Pharmacological studies on novel neurotensin mimetics: discovery of a pharmacologically unique agent exhibiting concentration-dependent dual effects as antagonist and agonist. AB - We report the development of two novel neurotensin mimetics (mimics 1 and 2). These compounds were rationally designed and synthesized according to the multiple template approach. We present results of experiments designed to define their pharmacological profiles. In radioligand binding assays with murine neuroblastoma clone N1E-115, we determined the equilibrium dissociation constants for these compounds at the neurotensin receptor. The Kd values for mimic 1 and mimic 2 were 3.3 microM and 1.9 microM, respectively. Functionally, both mimetics antagonized the neurotensin-stimulated production of cGMP, with Kd values in the low micromolar range. Interestingly, mimic 2 displayed a dualistic pharmacological profile, which was concentration dependent. At doses in the 10 100 microM range, mimic 2 became a full agonist, stimulating cGMP production in N1E-115 cells with an EC50 value of 19 microM. Furthermore, mimic 1 did not antagonize the cGMP response elicited by mimic 2. When the neurotensin receptor was desensitized with a neurotensin receptor agonist, mimic 2 failed to stimulate significant cGMP production. We propose that mimic 2 binds to a higher affinity site when acting as an antagonist and binds to a lower affinity and different site when acting as an agonist. Thus, mimic 2 would appear to represent a unique pharmacological tool to characterize the neurotensin receptor and its diverse binding sites in N1E-115 cells. PMID- 8246907 TI - Pharmacological characterization of stably transfected Na+/H+ antiporter isoforms using amiloride analogs and a new inhibitor exhibiting anti-ischemic properties. AB - A fibroblast mutant cell line devoid of Na+/H+ exchange was used to stably express cDNAs encoding the NHE1, NHE2, and NHE3 Na+/H+ antiporters. Pharmacological studies using amiloride and two of its 5-N-substituted derivatives, 5-N-dimethyl amiloride and 5-N-(methyl-propyl)amiloride (MPA), demonstrate that the NHE1 isoform is the ubiquitously expressed amiloride sensitive Na+/H+ antiporter (Ki of 0.08 microM for MPA), whereas the NHE2 and NHE3 isoforms exhibit a lower affinity for these inhibitors (Ki of 0.5 microM and 10 microM, respectively, for MPA) and are therefore likely to be members of the epithelial Na+/H+ exchanger's family. In addition, we have used this system to test a new Na+/H+ exchanger inhibitor possessing anti-ischemic properties on myocardial cells [(3-methylsulphonyl-4-piperidinobenzoyl) guanidine methanesulphonate]. This compound inhibits competitively NHE1 (Ki of 0.16 microM) with a much greater affinity than NHE2 and NHE3 (Ki of 5 microM and 650 microM, respectively) and therefore appears to be much more discriminative between these two classes of antiporter isoforms than the amiloride-related molecules. These results suggest an explanation for the observed difference of physiological effects between amiloride and HOE694, and identify this new inhibitor as a useful tool for studies of Na+/H+ exchange. PMID- 8246908 TI - Pharmacological characterization of alpha-bungarotoxin-sensitive acetylcholine receptors immunoisolated from chick retina: contrasting properties of alpha 7 and alpha 8 subunit-containing subtypes. AB - At least three subtypes of alpha-bungarotoxin-sensitive acetylcholine receptors (alpha Bgt-sensitive AChRs) exist in chick brain and retina. All may contain previously unknown structural subunits. One subtype contains alpha 7 subunits. Another contains alpha 8 subunits. A third contains both alpha 7 and alpha 8 subunits. In this article, we describe, for the first time, the pharmacological characterization of alpha 7 AChRs and alpha 8 AChRs immunoisolated from chick retina. Pharmacologically, the alpha 8 AChRs exhibit two classes of binding sites, the high affinity of which have higher affinity for most cholinergic ligands than do alpha 7 AChRs. These differences are most accentuated for ACh (approximately 5400-fold), decamethonium (approximately 1400-fold), 1,1,-dimethyl 4 phenylpiperazinium (approximately 200-fold), atropine (approximately 200-fold), nicotine (approximately 100-fold), and tetramethylammonium (approximately 100 fold). The alpha 8 AChR low affinity sites exhibit affinities that are similar but not identical to that of alpha 7 AChRs. Many of the pharmacological differences between the alpha 7 AChRs and alpha 8 AChRs can be attributed to the limited differences between the amino acid sequences of the N-terminal region of the alpha 7 and alpha 8 subunits because expressed alpha 7 homomers and alpha 8 homomers also exhibit these characteristic differences. PMID- 8246909 TI - Antihelicase action of DNA-binding anticancer agents: relationship to guanosine cytidine intercalator binding. AB - DNA-binding antibiotics such as intercalators, narrow groove binders, and other substances modify duplex DNA, making it an altered substrate for DNA helicases. The intercalators daunorubicin, actinomycin D, echinomycin, and elsamicin, the narrow groove binders distamycin and mithramycin, and the plant toxin teniposide, each representing a different chemical class, block SV40 large T antigen DNA helicase action with IC50 values ranging from 4 x 10(-8) to 2 x 10(-6) M. A partially purified human HeLa cell DNA helicase is also potently blocked by daunorubicin, distamycin, and teniposide. Because eukaryotic cells contain helicases of varying abundance, specificity, and type, this site of action for DNA-binding antibiotics may help explain antibiotic potency and specificity for DNA or RNA inhibition. The antihelicase effect of the antibiotic-double-stranded DNA complex may be central to the anticancer activities of these substances. An additional interesting correlation is the antihelicase action of DNA intercalating antibiotics and their DNA-binding preference for G-C base pair sites. The G-C base pair binding preference of the intercalating antibiotics may result from evolutionary selection because of the higher G-C binding stability, compared with A-T binding stability. The combination of the higher base pair stability at G-C regions and increased duplex DNA stability induced by intercalating antibiotic yields a total additive stability of the intercalator-G C base pair complex that resists helicase action. PMID- 8246910 TI - N-(phosphonacetyl)-L-aspartate synergistically enhances the cytotoxicity of 5 fluorouracil/interferon-alpha-2a against human colon cancer cell lines. AB - Recombinant interferon-alpha (IFN) enhances the cytotoxic effects of the fluorinated pyrimidine, 5-fluorouracil (5FU), against two human colon cancer cell lines. The aspartate transcarbamylase (ATCase) inhibitor, N-(phosphonacetyl)-L aspartate (PALA), was studied in combination with 5FU/IFN to determine whether further anti-pyrimidine effects would result in greater cytotoxicity. By median effects analysis PALA synergistically augmented the cytotoxic effects of 5FU/IFN against both human colon cancer cell lines. This occurred in the absence of any effects of 5FU/IFN on ATCase and without further potentiation of the PALA mediated inhibition of ATCase. To explore the mechanism by which this interaction occurred, detailed studies of pools of dNTPs were performed. Both 5FU/IFN and PALA/5FU/IFN treatments resulted in early (2-8 hr) depletion of pools of dTTP, but no effects on pools of dCTP. PALA had no effect on dTTP pools either alone or in the combination. In contrast, both PALA and PALA/5FU/IFN treatments resulted in later (12-24 hr) depletion of pools of dCTP. 5FU/IFN treatment had no effect on these pools. When pools of dCTP and dTTP were repleted by treatment with cytidine or thymidine, 20 microM, however, there was only partial reversal of cytotoxicity induced by 5FU/IFN + PALA, suggesting that the synergy observed did not result solely from a sequential anti-pyrimidine effect. The incorporation of 5FU into RNA was also studied; PALA enhanced the incorporation of [6-3H]5FU into RNA by 83-150%, but not into DNA, suggesting an alternative mechanism of drug interaction. PMID- 8246911 TI - Role of brain cytochrome P450 in regulation of the level of anesthetic steroids in the brain. AB - The role of brain cytochrome P450 (P450) in regulating the levels of the potent anesthetic steroid 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha-OH-DHP) has been investigated. By analogy with the elimination of androgen from its target tissues, we present evidence that it is 3 beta-hydroxy-5 alpha-pregnan-20-one (3 beta-OH-DHP) and not 3 alpha-OH-DHP that represents the major pathway for the formation of more polar metabolites and thus the elimination of the 5 alpha reduced metabolites of progesterone from target tissues. No polar metabolites were formed when 3 alpha-OH-DHP was incubated with microsomal fractions prepared from rat brain, but 3 beta-OH-DHP was hydroxylated at the 6 alpha- and 7 alpha positions. These 3 beta-diols were not formed to any detectable extent in the liver or kidney but were formed in prostate, pituitary, brain, and breast. The highest catalytic activity, 512 nmol of products formed/g of tissue/hr, was found in the prostate. The corresponding rates in the pituitary, brain, and breast were 71.9, 28.1, and 6.7 nmol/g/hr, respectively. These hydroxylations were confirmed to be P450-catalyzed reactions by solubilization of the P450 from prostate, brain, and breast microsomes and reconstitution of the catalytic activity with NADPH-P450 reductase (EC 1.6.2.4) and lipid. Because 5 alpha-androstane-3 beta,17 beta-diol (3 beta-Adiol) has been shown to be a good substrate for prostate and brain P450, competition experiments were performed to determine whether the same form of P450 is involved in the elimination of 3 beta-Adiol and 3 beta-OH-DHP in the brain. These two substrates competed with each other for metabolism in microsomal fractions and in reconstitution experiments with P450 extracted from the brain or prostate. To test the hypothesis that the hydroxylation of 3 beta-OH DHP represents a pathway for regulation of the level of 3 alpha-OH-DHP in the brain, the effect of inhibition of the hydroxylation of 3 beta-OH-DHP on the duration of 3 alpha-OH-DHP-induced anesthesia was examined. The nonanesthetic steroid 3 beta-Adiol was used as a competitive inhibitor of the metabolism of 3 beta-OH-DHP. The duration of anesthesia upon intravenous administration of 3 alpha-OH-DHP was increased by 33% when 3 beta-Adiol was coadministered.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246912 TI - Multiple sulfonylurea-sensitive potassium channels: a novel subtype modulated by dopamine. AB - In single channel patch-clamp recordings from freshly dissociated rat corpus striatum (caudate-putamen) neurons, the sulfonylurea drugs tolbutamide and glibenclamide caused a concentration-dependent blockade of a K+ channel that is activated by D2 dopamine receptor agonists. Tolbutamide was about 10-100 times more potent than glibenclamide, a rank-order potency opposite to that seen at previously described adenosine triphosphate-sensitive K+ channels. The channel also was poorly activated by diazoxide, which is a known opener of adenosine triphosphate-sensitive K+ channels. However, like adenosine triphosphate sensitive channels, it opened in the absence of dopaminergic agonist when the cells were treated with the metabolic inhibitor rotenone, indicating that channel openings occur under energy-depleting conditions. This suggests the existence of a novel, pharmacologically distinct class of sulfonyl-urea-sensitive K+ channels, regulated metabolically and also mediating dopaminergic neurotransmission. PMID- 8246913 TI - Cloning and expression of a human Ah receptor cDNA. AB - In this report, we describe the cloning and expression of a cDNA encoding a human Ah receptor, a basic/helix-loop-helix protein that mediates the toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin. A comparison of this human cDNA with a murine homologue (Ahb1 allele) indicates that the molecular mass variation observed between the receptors found in these two species results from hypervariability of amino acid sequences in the carboxyl termini (< 60% conserved over 450 amino acids). Differential usage of stop codons generates proteins with molecular masses that differ by 6 kDa. In contrast, the amino-terminal halves of these proteins are highly conserved and show 90% amino acid sequence identity. Northern blot analysis indicates that the human Ah receptor mRNA is expressed at its highest levels in placenta and is also highly expressed in lung, heart, pancreas, and liver, with lower levels of expression found in brain, kidney, and skeletal muscle. Expression of the human cDNA in a rabbit reticulocyte lysate system allowed functional analysis of ligand binding, agonist-induced and Ah receptor nuclear translocator-dependent DNA binding, and receptor stabilization by sodium molybdate. PMID- 8246914 TI - Cloning and characterization of the opossum kidney cell D1 dopamine receptor: expression of identical D1A and D1B dopamine receptor mRNAs in opossum kidney and brain. AB - Opossum kidney cells are an established epithelial cell line which is often studied as a physiological model system of renal proximal tubule function, and which has also been shown to possess dopamine receptors. To identify dopamine receptor subtypes present in renal tissue, as well as to explore the usefulness of opossum kidney cells for the study of D1 dopamine receptors and renal dopaminergic physiology, we have undertaken the cloning and characterization of the dopamine receptor expressed in this cell line. In the brains of rats and humans, two different subtypes of D1 dopamine receptors, D1A and D1B, have recently been characterized. The OK cell D1 receptor message is 4500 bp long and exhibits extensive homology with the rat and human D1A subtypes of dopamine receptors. Pharmacological experiments were performed on COS-7 cell membranes transiently transfected with this cDNA. Binding properties were compared with those reported for OK cell membranes, and comparison experiments were performed in parallel with the human D1A expressed transiently in the same system. Molecular techniques including Northern blotting, in situ hybridization, and RNase protection analysis were used to study the expression pattern of the OK cell D1 receptor message. Expression of both D1A and D1B subtypes was detected in both the opossum brain and the opossum kidney, however, the OK cell line expresses exclusively the D1A receptor subtype. PMID- 8246915 TI - Heterogeneous expression of alpha 1-adrenoceptor subtypes among rat nephron segments. AB - alpha 1-Adrenoceptor subtypes mediate many of the actions of the renal nerve, but their locations along the nephron are unknown. We investigated the distribution of alpha 1-adrenoceptor subtype mRNA and protein in rat proximal tubules and medullary thick ascending limbs (MTAL) using reverse transcription combined with polymerase chain reaction (PCR) and radioligand binding methods. Complementary primers were designed to span cDNA sequences in each of the third intracellular loops of the rat alpha 1B- and alpha 1D-adrenoceptors. Expression of the mRNA of alpha 1B- and alpha 1D-adrenoceptors was first detected in total RNA from whole rat kidney, and the PCR product identity was confirmed by sequencing. Endogenous expression of alpha 1B- and/or alpha 1D-adrenoceptor mRNA was then investigated in microdissected segments of the rat proximal convoluted tubule (S2 segments) and the MTAL. mRNA was reverse-transcribed directly from permeabilized microdissected segments and the resulting cDNA was subjected to PCR with the alpha 1-adrenoceptor primers. In proximal convoluted tubules, amplification of both alpha 1B- and alpha 1D-adrenoceptor mRNA was observed. In MTAL segments, only alpha 1D-adrenoceptor mRNA was detected. We also measured receptor protein using [3H]prazosin in saturation and competition binding experiments. Proximal tubular membranes contained 3.3-fold more alpha 1-adrenoceptor than did MTAL membranes (163 +/- 21 versus 49 +/- 3 fmol/mg of protein). When the alkylating agent chloroethylclonidine (CEC) (10 microM, 10 min) was used to define alpha 1 adrenoceptor subtypes, proximal tubules were found to contain primarily CEC insensitive (alpha 1A) sites (68 +/- 4%) and MTAL primarily CEC-sensitive sites (75 +/- 3%). Most [3H]prazosin binding sites (72 +/- 2%) in MTAL segments were also sensitive to the alkylating agent SZL-49, consistent with their identification as alpha 1D-adrenoceptors. In competition studies with the antagonists WB4101, 5-methylurapidil, and (+)-niguldipine, both high and low affinity sites were observed in proximal tubules. WB4101 interacted with only one site in MTAL membranes, intermediate in affinity between those sites found in proximal tubules. We conclude that reverse transcription-PCR is a useful method for demonstrating the expression of alpha 1-adrenoceptor subtypes in small amounts of tissue. Results from our experiments suggest that alpha 1A-, alpha 1B , and alpha 1D-adrenoceptors are all expressed in proximal tubules and that alpha 1D-adrenoceptors are the primary alpha 1-adrenoceptor subtype expressed in MTAL. The distinct anatomical distribution of each of these adrenoceptor subtypes suggests that they serve different functions in the kidney. PMID- 8246916 TI - Point mutation in the seventh hydrophobic domain of the human thromboxane A2 receptor allows discrimination between agonist and antagonist binding sites. AB - Thromboxane A2, a potent platelet agonist and vasoconstrictor, exerts its actions via specific G protein-coupled receptors. cDNAs encoding the full length thromboxane receptor have been isolated from human placenta mRNA by reverse transcriptase-polymerase chain reaction. An expression construct, under control of the cytomegalovirus promoter, was introduced into human embryonic kidney 293 cells. Membranes from transfected cells bound the thromboxane antagonist SQ29,548 and the agonist [15-(1 alpha,2 beta(5z)-3 alpha(1E,3S)-4 alpha)]-7-[3-(3-hydroxy 4-(p- iodophenoxy)-1-butenyl)-7-oxabicyclo[2,2,1]hept-2-yl]-5-heptenoic acid) with high affinities, and significantly more receptors were expressed in these cells, compared with platelet preparations. The putative seventh transmembrane segment is highly related in all cloned members of the eicosanoid receptor family and forms a critical portion of the ligand binding pocket for G protein-coupled receptors. Several point mutations in this segment were generated. Binding of SQ29,548 was virtually abolished in cells transfected with all the variant receptor constructs. However, one receptor variant (TxR-W299L), in which a tryptophan at position 299 was substituted for a leucine residue, allowed a definite discrimination between agonist and antagonist binding sites in competition and saturation binding experiments. An antibody directed toward the third intracellular loop of the thromboxane receptor was able to immunoprecipitate native thromboxane receptor in solubilized membranes from human erythroleukemia cells and transfected cells. PMID- 8246917 TI - Agonist-induced down-regulation and antagonist-induced up-regulation of m2- and m3-muscarinic acetylcholine receptor mRNA and protein in cultured cerebellar granule cells. AB - Cerebellar granule cells express m2- and m3-muscarinic acetylcholine receptors (mAChRs) and their corresponding mRNA with m3-mAChR being the predominant receptor subtype. After stimulation with the mAChR agonist, carbachol, m2- and m3 mAChR mRNA levels were decreased in a time- and concentration-dependent manner with the maximal down-regulation at 2 and 8 hr, respectively. Immunoprecipitation studies revealed that amounts of m2- and m3-mAChR protein also decreased at 8 and 24 hr, respectively. The carbachol-induced down-regulation of m3-mAChR mRNA was associated with a decrease in the transcription rate, but a substantial enhancement of the mRNA stability. Upon removal of carbachol after treatment for 8 hr, the levels of m3-mAChR mRNA and mAChR binding sites returned to their original values with a t1/2 of approximately 80 min and 6 hr, respectively. The carbachol-elicited loss of m2- and m3-mAChR mRNA was blocked by their corresponding receptor subtype-specific antagonists, AF-DX 116 (m2-selective) and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) (m3-selective), and was concurrent with an increase in c-fos mRNA levels. Exposure of granule cells to the nonselective mAChR antagonist, atropine, caused a time- and concentration dependent increase in the level of both m2- and m3-mAChR mRNA and mAChR binding sites. At 24 hr, immunoprecipitable m3-mAChR protein was predominantly increased. The atropine-induced up-regulation of m3-mAChR mRNA was concurrent with a marked enhancement of the mRNA stability and its transcription rate. The elevated levels of m3-mAChR mRNA and binding sites declined to their untreated values after the removal of atropine. Treatment with AF-DX 116 and 4-DAMP also produced an increase in the level of m2- and m3-mAChR mRNA and their corresponding immunoprecipitable receptor protein. These results demonstrate that the mAChR agonist and antagonist induce a down- and up-regulation of mAChR expression, respectively, through receptor-mediated mechanisms in cerebellar granule cells. Moreover, at least for m3-mAChR mRNA, the agonist- and antagonist-induced effects are reversible and associated with corresponding changes in the transcription rate of this receptor mRNA species. PMID- 8246918 TI - Multiple mechanisms for desensitization of A2a adenosine receptor-mediated cAMP elevation in rat pheochromocytoma PC12 cells. AB - To understand the regulation of A2a adenosine receptor (A2a-R) response, we examined the molecular mechanisms underlying the desensitization of A2a response in rat pheochromocytoma PC12 cells, which possess an A2a-R identical with the A2a receptor we recently cloned from rat brain. Prolonged exposure of PC12 cells to adenosine agonists significantly inhibited the response of the cells to subsequent stimulation with an A2a-selective adenosine agonist (CGS21680). No significant change in the number of binding sites and affinity for CGS21680 was observed in desensitized cells, nor did we find any significant change in the transcript level of A2a-R in cells pretreated with adenosine agonists. However, the basal adenylyl cyclase activity and the cyclase activities stimulated by adenosine agonists, by GTP gamma S, and by forskolin were reduced in desensitized cells. Prolonged exposure of PC12 cells to dibutyryl-cAMP did not significantly change either the basal or the adenosine agonist-evoked adenylyl cyclase activity. Therefore, elevation of cellular cAMP content is by itself not sufficient to produce the observed reductions of adenylyl cyclase activity with A2a desensitization. Inhibition of adenylyl cyclase activity in desensitized cells occurred after short-term (30 min) incubation with CGS21680 and could be blocked by the adenosine antagonist 8-cyclopentyl-1,3-dipropylxanthine. Gs alpha protein levels did not significantly change after a 30-min exposure to CGS21680. In contrast, long-term exposure (12-20 hr) of PC12 cells to adenosine agonists resulted in a slight further reduction of adenylyl cyclase activity and a consistent decline in the Gs alpha protein level. In addition, long-term incubation with adenosine agonists or with forskolin-enhanced phosphodiesterase (PDE) activity in the cytosolic and membrane fractions by 57 +/- 9% and 53 +/- 18%, respectively. Hydrolysis of cAMP was significantly faster in agonist desensitized cells than in control cells. PDE might therefore play an important role in desensitization of the A2a response in PC12 cells. Polymerase chain reaction-based analysis of the mRNA for A2a-R and A2b-R indicated that both A2a-R and A2b-R were present in PC12 cells; the A2b response was also diminished in A2a desensitized cells. Our data suggest that inhibition of adenylyl cyclase after short-term agonist treatment, down-regulation of Gs alpha protein level after long-term agonist treatment, and activation of PDE after long-term agonist treatment account for desensitization of the A2a-mediated response in PC12 cells. PMID- 8246919 TI - Expression of endothelin receptor subtypes in rabbit saphenous vein. AB - Recent investigations have revealed the presence of vasoconstrictory endothelin (ET)-B receptors in several tissues, including the rabbit saphenous vein (RSV). To determine the nature of the ET binding sites in RSV, radioligand-receptor binding studies with selective ligands and Northern analyses with probes from the ET-A and ET-B receptor cDNAs were conducted. ET-1 inhibited 125I-ET-1 binding to RSV in a monophasic manner, with an inhibition constant (Ki) of 0.08 +/- 0.02 nM and a slope factor of 0.9 +/- 0.1. ET-3 inhibition of 125I-ET-1 binding was biphasic, with 68% of the 125I-ET-1 binding sites being displaceable with a Ki value of 31 +/- 4 nM. The remaining 32% of the sites displayed high affinity for ET-3 (Ki = 0.2 +/- 0.1 nM). The ET-A-selective peptide BQ-123 inhibited 125I-ET-1 binding in a biphasic manner, with Ki values of 10.4 +/- 1.9 nM and 3.2 +/- 0.9 microM. The high affinity BQ-123 site comprised 70% of the binding sites, whereas the low affinity site comprised 30%. The correspondence of high affinity binding sites for BQ-123 and low affinity binding sites for ET-3 is consistent with the suggestion that 70% of the 125I-ET-1 binding sites in this tissue are ET-A receptors. To further investigate the nature of the ET-B binding sites in RSV, 125I-ET-3 competition binding experiments were conducted. ET-1 and BQ-123 inhibited 125I-ET-3 binding in RSV with Ki values of 40 +/- 7 pM and 7.2 microM, respectively, whereas inhibition curves for ET-3 and the ET-B receptor-selective agonist sarafotoxin S6c (S6c) were best fit to two-site models. Resultant Ki values for ET-3 and S6c were 50 pM (71%)/4 pM (29%) and 0.7 nM (71%)/95 nM (29%), respectively. Binding in RSV differed from that in rat cerebellum, where ET-3 and S6c inhibition of 125I-ET-3 binding was monophasic (Ki values of 70 pM and 1.1 nM for ET-3 and S6c, respectively). The presence of the nonhydrolyzable guanine nucleotide analog guanosine-5'-O-(3-thio)triphosphate (200 microM) did not affect 125I-ET-3 binding. Low stringency Northern analysis of RSV RNA with [alpha 32P]dCTP-labeled fragments from the ET-A or ET-B receptor cDNAs revealed similar hybridization patterns with both probes, with two resolved RNA species migrating at 4.7 and 1.8 kilobases.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8246920 TI - Identification of a 27-kDa high affinity phenylalkylamine-binding polypeptide as the sigma 1 binding site by photoaffinity labeling and ligand-directed antibodies. AB - The verapamil-like arylazide (-)-[3H]azidopamil specifically photoaffinity labeled two low molecular mass polypeptides, with apparent molecular masses of 22 and 27 kDa, in the endoplasmic reticulum of guinea pig liver, kidney, adrenal gland, and lung. It was recently shown that the 22-kDa polypeptide binds the anti ischemic phenylalkylamine (-)-[3H]emopamil and other anti-ischemic drugs with high affinity. We now provide evidence that the photolabeling of the 27-kDa polypeptide is blocked by nanomolar concentrations of sigma ligands [order of potency, haloperidol > pentazocine > 1,3-ditolylguanidine > dextromethorphan > (+)-SKF10,047]. The apparent affinities of these and other drugs closely corresponded to those for 1,3-[3H]ditolylguanidine-labeled sigma binding sites. Based on its high affinity for the (+)-enantiomer [but not the (-)-enantiomer] of SKF10,047 (Ki = 51 nM), pentazocine (Ki = 3 nM), and dextromethorphan (Ki = 30 nM), the (-)-[3H]azidopamil-labeled site on the 27-kDa polypeptide was classified as being of the sigma 1 subtype. Using antiphenylalkylamine antibodies, we developed a novel immunological detection method that allows the rapid and sensitive staining of the photolabeled 27-kDa polypeptide after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We conclude that the phenylalkylamines emopamil and azidopamil represent a novel class of sigma ligands, highly suitable for the further structural characterization of polypeptides carrying sigma 1 binding sites. PMID- 8246921 TI - Peptide alpha-amidation and peptidylglycine alpha-hydroxylating monooxygenase: control by disulfiram. AB - The final two steps in the biosynthesis of alpha-amidated bioactive peptides are catalyzed by peptidylglycine alpha-hydroxylating monooxygenase (PHM; EC 1.14.17.3) and peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL; EC 4.3.2.5). These enzymes are derived from the bifunctional precursor protein, peptidylglycine alpha-amidating monooxygenase. Because PHM is rate-limiting in peptide amidation and is copper-dependent, we examined the consequences of in vivo treatments with the copper-chelating drug disulfiram (Antabuse) on levels of alpha-amidated peptides and expression of PHM and PAL. Decreases in two amidated peptides (alpha-melanotropin and cholecystokinin) after disulfiram treatment were extremely pronounced outside the blood-brain barrier, with moderate decreases in the central nervous system. Unexpectedly, when assayed under optimal conditions in vitro, PHM activity was increased by disulfiram treatment, whereas PAL activity was unaltered. The increase in PHM activity in pituitary and atrium occurred within a few hours after the start of disulfiram treatment and was sustained up to 2 weeks after the cessation of treatment, whereas levels of alpha amidated peptides remained low. Northern and Western blot analyses demonstrated that disulfiram had no influence on levels of peptidylglycine alpha-amidating monooxygenase mRNA or protein. Thus, inhibition of alpha-amidation by disulfiram in vivo occurs despite an increased Vmax of PHM assayed in vitro. The increase in PHM activity may result from induction of a physiologic mechanism that normally regulates this rate-limiting enzyme. PMID- 8246922 TI - Regional distribution and expression modulation of cytochrome P-450 and epoxide hydrolase mRNAs in the rat brain. AB - In the present study, we developed a very sensitive, semiquantitative assay based on the reverse transcriptase-coupled polymerase chain reaction to measure, in a region-selective manner, mRNA expression patterns within the brain for microsomal epoxide hydrolase and several cytochrome P-450s (P-450s) known to be induced by prototypic agents in other tissues. The P-450s assessed included the polyaromatic hydrocarbon-inducible CYP1A1 and CYP1A2 systems, together with the phenobarbital inducible P-450s, CYP2B1, CYP2B2, CYP3A1, which were examined 18 hr after a single intraperitoneal dose of the respective inducing agents. Highly region specific patterns of expression were evident for P-450 mRNAs within the rat brain. In the control, uninduced brain, CYP1A1 mRNAs were readily detected in the striatum and in the hypothalamus, and to a lesser extent in the other regions examined. The regional pattern of expression was similar for CYP1A2; however, a major difference was noted in the olfactory bulbs, characterized by a relatively high level of CYP1A2 mRNA but correspondingly low levels of CYP1A1. Within the brain regions examined, the highest content of CYP2B1 and CYP2B2 mRNAs were present in the striatum and in the cerebellum, whereas CYP3A1 levels varied only slightly across the respective regions. In contrast to the P-450s, microsomal epoxide hydrolase mRNAs were expressed at relative homogeneous amounts throughout the brain. beta-Naphthoflavone markedly increased the CYP1A1 and CYP1A2 mRNA contents of each brain region investigated, although this agent did not affect levels of epoxide hydrolase. At 18 hr post-treatment with phenobarbital, an optimal time period for hepatic induction, brain expression was characterized by a complex pattern of effects, with increased levels noted for CYP2B1 mRNA content in the medulla oblongata, midbrain, and cortex, but decreased contents measured in the cerebellum, the hypothalamus, and the striatum. In each of these respective regions, CYP2B2 content was profoundly decreased whereas epoxide hydrolase expression was slightly increased by the same treatment. These results establish that the central nervous system actively expresses a number of different biotransformation gene products in a regional specific and inducer dependent manner, and suggest that for tissues exhibiting low regenerative capacity, like the brain, such reactions are likely to be of critical toxicological significance. PMID- 8246923 TI - Growth hormone and vitamin A induce P4502C7 mRNA expression in primary rat hepatocytes. AB - The effects of growth hormone (GH) and retinoids on P4502C7 mRNA levels were investigated in cultured primary hepatocytes from normal female rats. Northern blot analysis of total nucleic acids from hepatocytes maintained in culture for 90 hr showed low basal levels of P4502C7 mRNA, which were marginally increased after continuous treatment with GH. Retinal treatment gave a slightly higher induction than GH, whereas treatment with all-trans retinoic acid alone or with GH in combination with retinol induced P4502C7 mRNA to levels about one-third of those in normal female rat liver. The effects of retinoids on P4502C7 mRNA were dependent on both the dose and type of retinoid used. All-trans retinoic acid produced a saturable dose-response curve with a 50% maximal induction of P4502C7 mRNA at 1.5 microM. The isomer 9-cis retinoic acid showed a dose-dependent activation of P4502C7 mRNA similar to that of all-trans retinoic acid. Retinol gave a 50% maximal response at approximately 5 microM. In the presence of GH, the induction of P4502C7 mRNA appeared additive to the effect of retinol at all concentrations used and to all-trans retinoic acid at concentrations up to 1 microM. As determined by a quantitative solution hybridization assay, P4502C7 mRNA levels were induced 3-fold by GH, 5-fold by retinol, and 19-fold by all trans retinoic acid. In the presence of GH, P4502C7 was induced 8-fold by retinol, whereas the induction by saturating concentration of all-trans retinoic acid showed no significant additional effect of GH. The importance of vitamin A for the expression of P4502C7 in vivo was confirmed by the low abundance of P4502C7 mRNA in vitamin A-deficient animals as compared with vitamin A-adequate control rats. Nuclear run-on experiments performed in cultured primary hepatocytes showed that both GH and retinoic acid exert their effects at the transcriptional level. We conclude that both GH and retinoids can induce P4502C7 mRNA in rat liver hepatocytes, retinoic acid being the dominant inducer. PMID- 8246924 TI - [Expression of the human alpha-1-antitrypsin gene in Escherichia coli]. AB - The cDNA, containing the complete human alpha-1-antitrypsin (AT) sequence starting from codon 2, was used to construct bacterial strains producing AT. The fusion protein was obtained by junction of the AT cDNA to the fragment of an Escherichia coli ompF gene. We have also modified the AT cDNA's 5'-terminal part to construct DNAs containing ATG-codon and cDNA sequences starting from codons 1 or 2. These DNAs were inserted into E. coli expression vectors pBR322/trpII-8 and pKK223-3 that allow transcription from efficient trp- and tac-promoters. This construction resulted in the induction of a 46 kDa protein. The polypeptide produced was recognized by an antiserum raised against human alpha-1-antitrypsin protein. Truncation of the gene at its 5'-end or synthesis as a fusion OmpF-AT protein increases expression up to 10-fold, to a level of approximately 1%. On the contrary, no dependence on the promoter type has been observed. Physical properties of the recombinant proteins are discussed. PMID- 8246925 TI - [Proteins, interacting with the promotor of the immunoglobulin kappa gene]. AB - Two octanucleotide binding protein factors (mOct-1 and mOct-2A) are detected in nuclear extracts of lymphoid NS/0 cell line. These proteins, corresponding to human Oct-1 and Oct-2A nuclear factors, were purified by ion exchange and heparin agarose chromatography. With retardation assay technique it was shown that mOct 2A factor at all steps of purification contains tightly bound protein, dissociating in the presence of a nonspecific native DNA. This octa-protein associated factor (OAF) has low affinity to DNA and is not able to bind specifically the promoter of kappa-gene. However it interacts with DNA together with mOct-2A in highly specific manner but does not form complexes with mOct-1 and mOct-2B factors. Since OAF can be attached specifically only to mOct-2A (which play an essential role in initiation of kappa-gene transcription) this attachment may be considered as necessary for its recognition as a specific transcription factor. The ability of OAF like alpha TIF factor (specifically interacting with Oct-1) to discriminate between the Oct proteins, recognizing the same cis-acting oct-elements, shows how the non-DNA-binding transcription factors can influence differentially the transcription by two proteins binding to the same DNA sequences. PMID- 8246926 TI - [Standardization of a procedure for automatic determination of protein secondary structure from their circular dichroism spectra with respect to "aromatics"]. PMID- 8246927 TI - [Inhibition of reproduction of the human immunodeficiency virus by sense and antisense oligodeoxynucleotides]. AB - Inhibitory effects on human immunodeficiency virus (HIV) reproduction on lymphoid cell line MT-4 were characterized for antisense and sense oligodeoxynucleotides. It was established that antisense oligonucleotide pCGTAGTTCGTCGAGGTCCGT (MP-20) (ID50 = 0.1 microM) is a more effective HIV inhibitor than the previously described pTGGCGTACTCACCAGTCGCCGC (DSS-22) (ID50 = 4.7 microM) and pTTTTTTTTTTTTTTTT (PA-16) (ID50 = 8.0 microM). A sense oligonucleotide pGCATCAAGCAGCTCCAGGCA (PM-20) (ID50 = 0.5 microM) complementary to the region of the start of translation of the open reading frame on the (+)-chain virus DNA was also investigated. Specificity of the anti-HIV-I action of oligonucleotides was confirmed by experiments with HIV-II. PMID- 8246928 TI - [Accessible surface and intramolecular mobility of proteins: study by method of tritium planigraphy]. AB - The method of tritium planigraphy is used for determination of the accessible surface of a globular protein--lysozyme--and the accessibility of particular types of amino acid residues as a function of temperature in the range of 77-193 K. Protein powder with humidity 10 +/- 1% was used. As the temperature is changed from 77 to 160 K for all types of amino acid residues was obtained decreasing of inclusion of tritium label. All types of amino acid residues may be divided on the three groups: I. Accessibility rises under increasing temperature from 160 to 293 K (K, R, H, P, L); II. Accessibility not depends or slightly increases with the growth of temperature (C, V, A, I, Y, F); III. Accessibility strongly increases in the range of the temperature 260-293 K (S, T, G, D + N, E + Q). The reason of "cold denaturation" is perhaps the difference of behaviour of structural water molecules. Under increasing of temperature from 160 to 293 K change of accessibility was explained the growing of intramolecular flexibility of molecule. Under transition from 160 to 77 K for all types of residues is observed sensible change of spatial structure of the protein, which cannot be explained by only participation of dynamical characteristics. PMID- 8246929 TI - [2'-Deoxynucleoside-5'-triphosphate, modified via the carbohydrate and triphosphate residues, in the DNA synthesis reaction]. AB - We have investigated the substrate properties of deoxyribonucleoside 5' triphosphate analogues, modified in the carbohydrate and triphosphate moieties, in DNA synthesis catalyzed by different DNA polymerases and reverse transcriptases. It was shown that (3'-azido-2',3'-dideoxythymidine-5'-O methylenephosphonate) diphosphate, (3'-azido-2',3'-dideoxythymidine 5'-phosphate) dibromomethylenediphosphonate, (3'-azido-2',3'-dideoxythymidine 5'-phosphate) phosphonoacetate terminate DNA synthesis catalyzed by reverse transcriptases. (2' Deoxythymidine 5'-phosphate) phosphonoacetate displays substrate properties for DNA polymerase beta, different reverse transcriptases, terminal deoxynucleotidyl transferase, but not for DNA polymerase alpha, Klenow's fragment DNA polymerase I. The Km value for this substance in DNA synthesis reactions catalyzed by reverse transcriptases was two of orders magnitude higher than that for native 2' deoxythymidine 5'-triphosphate. PMID- 8246930 TI - [Short repeats and variability in the smooth noncoding area of human mitochondrial DNA]. AB - Major non-coding region of human mitochondrial DNA (mtDNA) (1122 bp) was assessed using the method of complexity analysis of genomes. The ACT, TCA, AGT and TGA motifs (AST-repeats) were shown to form short repeats as well as more complex block structures. These motifs are intrinsic for regulatory sequences of DNA of procaryotic and eucaryotic genes. ACT-repeats based blocks happen to be the most variable parts of the region studied too. Each inherited type of mtDNA is proposed to be a pattern of short repeats arranged with the regard to their symmetry, complementarity and alternativeness thus forming block DNA structures. The existence of similar structures may be possible due to the variability of nucleotide sequences more pronounced in the blocks of repeats of major non-coding region of human mtDNA. PMID- 8246931 TI - [Study of the structure of Escherichia coli RNA polymerase and its complex with the lacUV5-promotor using protein-protein and DNA-protein crosslinks, formed by formaldehyde]. AB - The protein-protein and DNA-protein crosslinks produced by formaldehyde were used to investigate the intersubunit and subunit-DNA interactions for free RNA polymerase and for an open complex of RNA polymerase with the lacUV5 promoter. In both cases the contacts between beta,beta' and beta', sigma subunits were observed, while there were no contacts between beta and sigma subunits. Only one of beta or beta' subunits and a sigma subunit crosslink to promoter DNA. We have chosen the conditions for fixing the RNA polymerase-DNA complexes on different stages of transcription initiation. The possibility to use limited fixation with low concentrations of formaldehyde to study specific DNA-protein interactions was shown. PMID- 8246932 TI - [Heterologous expression of functionally active human cytochrome P-450. I. Synthesis of cytochrome P-450IID6 by transformed Saccharomyces cerevisiae cells]. AB - Functionally active human cytochrome P-450IID6 was expressed in Saccharomyces cerevisiae cells transformed with yeast expression vectors containing a fragment of the 2 mu-plasmid and cytochrome P-450 cDNA. Transcription was regulated by yeast acid phosphatase PHO5 promoter or chimeric GAL10-CYC1 promoter. Induced yeast cells synthesized cytochrome P-450 that was revealed by the optical absorption of the reduced CO form. Intact yeast cells were capable to metabolize the drug sparteine forming biotransformation products usually found in human. Expression of the human cytochromes P-450 seems to be a promising route to investigate new drugs and to screen genotoxic compounds. PMID- 8246933 TI - [Cloning of the gene for thermostable Thermus aquaticus YT1 DNA polymerase and its expression in Escherichia coli]. AB - Using the phasmid vector pSL5, the genomic DNA fragment of T. aquaticus YT1 which contained the thermostable DNA polymerase (Taq-polymerase) gene was cloned. The BglII fragment of this genome locus was subcloned in the BamHI site of the pUC19 plasmid. To optimize the Taq-polymerase gene expression in E. coli cells, the gene was cloned in the correct reading frame regarding the initiation ATG codon of the pPR-TGATG-1 expression vector. The gene expression in this vector was controlled by the phage lambda PR promoter and the temperature-sensitive phage lambda repressor. We used PCR to amplify the short 5'-end fragment of the Taq polymerase gene coding for the part into which an artificial SacI site was introduced. This site has been used for cloning the PCR product into the pPR TGATG-1 vector, and the missing gene part was cloned into the KpnI site of the PCR product from the natural cloned gene. The cells of the E. coli PVG-A1 strain, which was obtained in the end, expressed efficiently the Taq-polymerase gene at the nonpermissive temperature. The content of the recombinant Taq-polymerase in the cells was about 1-2% of total proteins. The purified nearly homogeneous Taq polymerase amplified efficiently in the PCR DNA fragments up to 5.5 kb long and was useful in DNA sequencing the by Sanger method. The half-life of the purified Taq polymerase was about 60 min at 95 degrees C, it was active for at least 65 standard PCR circles. The specific activity of recombinant enzyme preparations was about 180-200,000 units per mg of protein. The E. coli PVG-A1 strain enables one to isolate up to 500,000 units of purified enzyme from 2 l of bacterial culture. PMID- 8246934 TI - [Biochemical features of protein matrix M1 of the influenza C virus]. AB - Influenza viruses A, B, and C belonging to Orthomyxoviridae comprise an internal ribonucleoprotein (RNP) and an outer lipoprotein envelope with surface spike glycoproteins and the M1 protein matrix. The lipoprotein envelope and spike glycoproteins are solubilized by nonionic detergent treatment in a pH-independent manner. In contrast, disassembly of the M1 protein matrix appears to depend on pH. Treatment of influenza C viruses with nonionic detergent in neutral or alkaline medium (pH 9.0-7.2) results in disintegration of the virion M1 matrix and leads to a significant release of RNP free of the M1 protein. In acidic medium (pH 6.0-5.0) the M1 matrix fails to be removed and the viral core-like complex of RNP along with the M1 matrix cover is released. Since influenza A and B viruses were characterised by acid-dependent disassembly of the virion M1 matrix, influenza C viruses seem to be more resemble the paramyxoviruses, which also show a neutral-alkaline pH dependence of the matrix disintegration. These observations suggest that uncoating of influenza C viruses and paramyxoviruses in target cells may have similar events. PMID- 8246935 TI - [Reconstruction of a sequenced sequence using results of stacked hybridization with an oligonucleotide matrix]. AB - The efficiency of additional rounds of continuous stacking hybridization in DNA sequence reconstruction by hybridization with oligonucleotide matrix (SHOM) is considered. At first, DNA is hybridized with a matrix of oligonucleotides of the length L. The overlapping of the tuples which have formed perfect duplexes with the DNA, on the one hand, has enabled us to reconstruct unambiguously a part of the sequence, and on the other hand, suggested a modified scheme for reconstructing the remaining part. Then the additional hybridizations should be carried out in the presence of shorter oligonucleotides of the length l, that are able to form perfect duplexes of the length L + l in continuous hybridization closely to matrix tuples. In this case the stability of a duplex consisting of l tuples and long DNA is enhanced due to stacking interaction. The information obtained about the succession of L + l sites considerably increases the efficiency of reconstruction, which can eventually reach the efficiency of a matrix consisting of (L + l)-tuples. We propose here an algorithm for compiling such a set of l-tuples to be added that the number of additional hybridizations can be appreciably diminished. For an octanucleotide matrix and different sets of pentanucleotides to be used for continuous stacking hybridization, the length of unambiguously sequenced DNA undergoes a rise from 200 to some thousands bp. PMID- 8246936 TI - [Study of the conformational and intradynamic changes of phage lambda DNA molecules by laser correlation spectroscopy]. AB - Conformational changes of lambda phage DNA macromolecules were investigated by dynamic light scattering (DLS) method. It was shown that the persistence length of B-form DNA in some conditions could could decline to approximately 12 nm. When studying the internal motions in the linear form of phage DNA macromolecule, a deviation was observed of the experimental angular dependence of the first cumulant gamma from theoretical predictions for the gamma in Rouse-Zimm model including hydrodynamic interactions. This deviation appeared in the region qRg >> 1, where Rg is gyration radius and q is momentum vector. The first cumulant deviation to higher frequencies is observed only for B-form DNA. Experimental behaviour of the first cumulant gamma for the A-form DNA coincides with the theoretical one in the Rouse-Zimm model. PMID- 8246937 TI - [The Z-form of DNA. Nonmonotonous change in stability with increase in ionic strength]. AB - To specify the stability area of the Z-DNA conformation as a function of ambient conditions (ionic strength, temperature, water activity) a 3-D phase diagram for the (B,Z) equilibrium as exemplified by poly(dG-5ethyldC) has been constructed in experiment. Its main peculiarity proves to be the nonmonotonic change in stability of the Z form with ionic strength, with a minimum at 0.1 M NaCl, i.e. the increase or decrease in salt concentration from the above value leads to stabilization of the Z form relatively to the B form. The shape of the phase diagram section (temperature, ionic strength) is indicative of the existence of two B-type conformations, equilibrium between them being unaffected by ionic strength though being sensitive to temperature. The B1 form is stable at t < 30 degrees C while the B2 one--at t > 30 degrees C. Using the general 3-D phase diagram together with a polyelectrolyte theory of the B-Z transition, thermodynamic parameters were calculated for the B1-Z, B2-Z, and B1-B2 conformational shifts as well as the B/Z junction energy. The totality of the obtained results with poly(dG-5ethyldC) and those earlier obtained by us with poly(dG-5methyldC) is in full qualitative and quantitative agreement with the polyelectrolyte model, and at the same time it makes unlikely the explanation suggested by others for the low-ionic Z to B transition as the one totally stipulated by a metal ion contamination. PMID- 8246938 TI - [Sex dimorphism in expression of alternative variants of mRNA for the growth hormone receptor in rat liver]. AB - Two alternative variants of 5'-untranslated sequence of the rat growth hormone receptor (GHR) mRNA were previously described. These variants of mRNA appear to be produced by the splicing of primary transcripts initiated from alternative promoters. We employed the ribonuclease protection assay to reveal these two variants of the GHR mRNA in RNA preparations from the liver of male, normal female, and pregnant female rats. All tissues examined contain approximately the same amount of the GHR mRNA with type II 5'-untranslated sequence. To the contrary, the content of type I variant shows very pronounced sexual differences; its level is very low in the male liver, approximately equal to that of the type II variant in the female liver, and further increases in the pregnant female liver. These data suggest that the higher content of the GHR observed in the female liver and its elevation during gestation result from the accumulation of "female specific" type I variants of the GHR mRNA. PMID- 8246939 TI - [The hierarchy of complexes and compact structures of trivaline with nucleic acids. I. Interaction of trivaline with double-stranded DNA]. AB - It was shown that trivaline-poly(U) complexes have no appreciable fluorescence at any peptide concentration if the polymer concentration is less than critical. By electron microscopy it was shown that dsDNA molecules undergo a second step of compactization if polymer and peptide concentrations are high enough. The diameter of this rod-like particles is over 40 nm. If the polymer concentration is less than critical, dsDNA molecules form with trivaline extended structures of dissimilar morphology. We propose a scheme of trivaline-dsDNA complexes and compact structure formation, beginning from a dimer complex. It is consistent with the results of equilibrium dialysis, fluorescence, electron microscopy, flow linear and circular dichroism measurements. PMID- 8246940 TI - [The mechanism of replicative and post-replicative DNA methylation as a generator of mutations in a cell]. AB - The functional role of enzymatic DNA methylation in eukaryotes is still obscure even 45 years after its discovery. In the present paper the analysis of various aspects of DNA methylation has been made from the point of view of a common functional model considering the system as a generator of 5mC-->T mutations in the cell. The mechanism and consequences of the reactions have been described in detail, from the 5mC residue deamination during replicative DNA methylation and repair of G.T-mispairs to production of hemimethylated sites in DNA and their postreplicative methylation. It was shown that the loss of the most part of the 5mC residues from DNA during the lifespan proceeds with aging both of organism's tissues and of cell cultures. Evidence was obtained that this is the result of gradual accumulation of 5mC-->T + C transitions in the genome during each cell division. Such substitutions take place 10 times more often in methylated sites *CG than in any other sites of DNA. They disproportionally contribute to general mutagenesis and may be a cause of many hereditary human diseases. The conclusion has been made that DNA methylation can be considered as a genetically programmed mechanism for accumulating mutations with aging. PMID- 8246941 TI - [Modular evolution of Bsp-repeats: formation of subrepeats and monomers preceded the divergence of four species of Canidae]. AB - Determination of two nucleotide sequences of satellite-like DNA (Bsp-repeats) segments from the raccoon-like dog genome and analysis of these and three other homologous sequences from distantly related genomes of Canidae (silver fox, domestic dog, and gray fox) show that all these DNA fragments consist of five types of subrepeats (A, B, C, D, and E) and two types of monomers-ACB and ACED. Raccoon-like dog repeated segment of 1620 bp is composed of both type of monomers: ACBACED. Different monomers of the same type could be distinguished by subtypes of subrepeats. Building of a cladogram of 18 subrepeats and analysis of the structures of monomers and higher-order repeated units allowed us to reveal probable evolution events which have determined the hierarchical order in Bsp repeat organization. Approximate periods of time for these evolution events were calculated from the number of substitutions per branch. Five main types of subrepeats and two types of monomers have been generated and propagated through several steps before divergence of the main phylogenetic trees of Canidae. Combinatorial variability appears to play a fundamental role in forming the hierarchical layers in Bsp-repeats structure. PMID- 8246942 TI - Dominant-negative mutants of platelet-derived growth factor revert the transformed phenotype of human astrocytoma cells. AB - Malignant astrocytoma is the most common primary human brain tumor. Most astrocytomas express a combination of platelet-derived growth factor (PDGF) and PDGF receptor which could close an autocrine loop. It is not known whether these autocrine loops contribute to the transformed phenotype of astrocytoma cells or are incidental to that phenotype. Here we show that dominant-negative mutants of the PDGF ligand break the autocrine loop and revert the phenotype of BALB/c 3T3 cells transformed by the PDGF-A or PDGF-B (c-sis) gene. Then, we show that these mutants are selective in that they do not alter the phenotype of 3T3 cells transformed by an activated Ha-ras or v-src gene or by simian virus 40. Finally, we show that these mutants revert the transformed phenotype of two independent human astrocytoma cell lines. They have no effect on the growth of human medulloblastoma, bladder carcinoma, or colon carcinoma cell lines. These observations are consistent with the view that PDGF autocrine loops contribute to the transformed phenotype of at least some human astrocytomas. PMID- 8246943 TI - A common element involved in transcriptional regulation of two DNA alkylation repair genes (MAG and MGT1) of Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae MAG gene encodes a 3-methyladenine DNA glycosylase that protects cells from killing by alkylating agents. MAG mRNA levels are induced not only by alkylating agents but also by DNA-damaging agents that do not produce alkylated DNA. We constructed a MAG-lacZ gene fusion to help identify the cis-acting promoter elements involved in regulating MAG expression. Deletion analysis defined the presence of one upstream activating sequence and one upstream repressing sequence (URS) and suggested the presence of a second URS. One of the MAG URS elements matches a decamer consensus sequence present in the promoters of 11 other S. cerevisiae DNA repair and metabolism genes, including the MGT1 gene, which encodes an O6-methylguanine DNA repair methyltransferase. Two proteins of 26 and 39 kDa bind specifically to the MAG and MGT1 URS elements. We suggest that the URS-binding proteins may play an important role in the coordinate regulation of these S. cerevisiae DNA repair genes. PMID- 8246945 TI - Architecture of the maize mitochondrial atp1 promoter as determined by linker scanning and point mutagenesis. AB - Plant mitochondrial promoters are poorly conserved but generally share a loose consensus sequence spanning approximately 17 nucleotides. Using a homologous in vitro transcription system, we have previously shown that an 11-nucleotide sequence within this region comprises at least part of the maize mitochondrial atp1 promoter (W. Rapp and D. Stern, EMBO J. 11:1065-1073, 1992). We have extended this finding by using a series of linker-scanning and point mutations to define the atp1 promoter in detail. Our results show that mutations at positions 12 to +5, relative to the major transcription start site, can decrease initiation rates to between < 10 and 40% of wild-type levels. Some mutations, scattered throughout this region, have lesser effects or no effect. Taken together, our data suggest a model in which the atp1 promoter consists of a central domain extending from -7 to +5 and an upstream domain of 1 to 3 bp that is centered around -11 to -12. Because many mutations within this promoter region are tolerated in vitro, the maize atp1 promoter is distinct from the highly conserved yeast mitochondrial promoters. PMID- 8246944 TI - The ionizing radiation-induced replication protein A phosphorylation response differs between ataxia telangiectasia and normal human cells. AB - Replication protein A (RPA), the trimeric single-stranded DNA-binding protein complex of eukaryotic cells, is important to DNA replication and repair. Phosphorylation of the p34 subunit of RPA is modulated by the cell cycle, occurring during S and G2 but not during G1. The function of phosphorylated p34 remains unknown. We show that RPA p34 phosphorylation is significantly induced by ionizing radiation. The phosphorylated form, p36, is similar if not identical to the phosphorylated S/G2 form. gamma-Irradiation-induced phosphorylation occurs without new protein synthesis and in cells in G1. Mutation of cdc2-type protein kinase phosphorylation sites in p34 eliminates the ionizing radiation response. The gamma-irradiation-induced phosphorylation of RPA p34 is delayed in cells from ataxia telangiectasia, a human inherited disease conferring DNA repair defects and early-onset tumorigenesis. UV-induced phosphorylation of RPA p34 occurs less rapidly than gamma-irradiation-induced phosphorylation but is kinetically similar between ataxia telangiectasia and normal cells. This is the first time that modification of a repair protein, RPA, has been linked with a DNA damage response and suggests that phosphorylation may play a role in regulating DNA repair pathways. PMID- 8246946 TI - Signaling activity of transforming growth factor beta type II receptors lacking specific domains in the cytoplasmic region. AB - The transforming growth factor beta (TGF-beta) type II receptor (T beta R-II) is a transmembrane serine/threonine kinase that contains two inserts in the kinase region and a serine/threonine-rich C-terminal extension. T beta R-II is required for TGF-beta binding to the type I receptor, with which it forms a heteromeric receptor complex, and its kinase activity is required for signaling by this complex. We investigated the role of various cytoplasmic regions in T beta R-II by altering or deleting these regions and determining the signaling activity of the resulting products in cell lines made resistant to TGF-beta by inactivation of the endogenous T beta R-II. TGF-beta binding to receptor I and responsiveness to TGF-beta in these cells can be restored by transfection of wild-type T beta R II. Using this system, we show that the kinase insert 1 and the C-terminal tail of T beta R-II, in contrast to the corresponding regions in most tyrosine kinase receptors, are not essential to specify ligand-induced responses. Insert 2 is necessary to support the catalytic activity of the receptor kinase, and its deletion yields a receptor that is unable to mediate any of the responses tested. However, substitution of this insert with insert 2 from the activin receptor, ActR-IIB, does not diminish the ability of T beta R-II to elicit these responses. A truncated T beta R-II lacking the cytoplasmic domain still binds TGF-beta, supports ligand binding to receptor I, and forms a complex with this receptor. However, TGF-beta binding to receptor I facilitated by this truncated T beta R-II fails to inhibit cell proliferation, activate extracellular matrix protein production, or activate transcription from a promoter containing TGF-beta responsive elements. We conclude that the transcriptional and antiproliferative responses to TGF-beta require both components of a heteromeric receptor complex that differs from tyrosine kinase receptors in its mode of signaling. PMID- 8246947 TI - Epidermal growth factor induces phosphorylation of extracellular signal-regulated kinase 2 via multiple pathways. AB - Expression of p21rasAsn-17, a dominant negative mutant of p21ras that blocks p21ras activation by growth factors, inhibits activation of extracellular signal regulated kinase 2 (ERK2) by insulin and platelet-derived growth factor in rat-1 cells [A. M. M. de Vries-Smits, B. M. T. Burgering, S. J. Leevers, C. J. Marshall, and J. L. Bos, Nature (London) 357:602-604, 1992]. Here we report that expression of p21rasAsn-17 does not abolish epidermal growth factor (EGF)-induced phosphorylation of ERK2 in fibroblasts. Since EGF activates p21ras in these cells, this indicates that EGF induces a p21ras-independent pathway for the phosphorylation of ERK2 as well. We investigated whether activation of protein kinase C (PKC) or increase in intracellular calcium could be involved in p21ras independent signaling. In rat-1 cells, inhibition of either PKC, by prolonged 12 O-tetradecanoylphorbol-13-acetate (TPA) pretreatment, or calcium influx, by ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) pretreatment, did not abolish EGF-induced ERK2 phosphorylation. However, a combined inhibition of both p21ras and calcium influx, but not PKC, resulted in a complete inhibition of EGF-induced ERK2 phosphorylation. In contrast, in Swiss 3T3 cells, inhibition of both p21ras activation and TPA-sensitive PKC, but not calcium influx, inhibited EGF-induced ERK2 phosphorylation. These results demonstrate that in fibroblasts, EGF induces alternative pathways of ERK2 phosphorylation in a cell-type-specific manner. PMID- 8246948 TI - Characterization of quail Pax-6 (Pax-QNR) proteins expressed in the neuroretina. AB - After differential screening of a cDNA library constructed from quail neuroretina cells (QNR) infected with the v-myc-containing avian retrovirus MC29, we have isolated a cDNA clone, Pax-QNR, homologous to the murine Pax-6, which is mutated in the autosomal dominant mutation small eye of mice and in the disorder aniridia in humans. Here we report the characterization of the Pax-QNR proteins expressed in the avian neuroretina. From bacterially expressed Pax-QNR peptides, we obtained rabbit antisera directed against different domains of the protein: paired domain (serum 11), domain between the paired domain and homeodomain (serum 12), homeodomain (serum 13), and carboxyl-terminal part (serum 14). Sera 12, 13, and 14 were able to specifically recognize five proteins (48, 46, 43, 33, and 32 kDa) in the neuroretina. In contrast to proteins of 48, 46, and 43 kDa, proteins of 33 and 32 kDa were not recognized by the paired antiserum (serum 11). Paired less and paired-containing proteins exhibited the same half-life (6 h) and were phosphorylated mostly on serine residues. Immunoprecipitations performed with subcellular fractions of neuroretinas showed that the paired-containing proteins were located in the nucleus, whereas the 33- and 32-kDa proteins were found essentially in the cytoplasmic compartment. However, immunofluorescence experiments performed after transient transfections showed that p46 and p33/32 were also located in vivo into the nucleus. Thus, the Pax-QNR/Pax-6 gene can produce proteins with two DNA-binding domains as well as proteins containing only the DNA-binding homeodomain. PMID- 8246949 TI - Independent regions of adenovirus E1A are required for binding to and dissociation of E2F-protein complexes. AB - The transcription factor E2F is present in independent complexes with the product of the retinoblastoma susceptibility gene, pRB, and a related gene product, p107, in association with the cyclin A-cdk2 or the cyclin E-cdk2 kinase complex. pRB and p107 can negatively regulate E2F activity, since overexpression of pRB or p107 in cells lacking a functional pRB leads to the repression of E2F activity. The products of the adenovirus E1A gene can disrupt E2F complexes and result in free and presumably active E2F transcription factor. The regions of E1A required for this function are also essential for binding to a number of cellular proteins, including pRB and p107. Through the use of a number of glutathione S transferase fusion proteins representing different regions of E1A, as well as in vivo expression of E1A proteins containing deletions of either conserved region 1 (CR1) or CR2, we find that CR2 of E1A can form stable complexes with E2F. E1A proteins containing both CR1 and CR2 also associate with E2F, although the presence of these proteins results in the release of free E2F from its complexes. In vitro reconstitution experiments indicate that E1A-E2F interactions are not direct and that pRB can serve to facilitate these interactions. Complexes containing E1A, p107, cyclin A, and E2F were identified in vivo, which indicates that E1A may associate with E2F through either p107 or pRB. Peptide competition experiments demonstrate that the pRB-binding domain of the human E2F-1 protein can compete with the CR1 but not CR2 domain of E1A for binding to pRB. These results indicate that E1A CR1 and E2F-1 may bind to the same or overlapping sites on pRB and that E1A CR2 binds to an independent region. On the basis of our results, we propose a two-step model for the release of E2F from pRB and p107 cellular proteins. PMID- 8246950 TI - Induction of RNA editing at heterologous sites by sequences in apolipoprotein B mRNA. AB - An RNA editing mechanism modifies apolipoprotein B (apo-B) mRNA in the intestine by converting cytosine at nucleotide (nt) 6666 to uracil. To define the sequence requirements for editing, mutant apo-B RNAs were analyzed for the ability to be edited in vitro by enterocyte extracts. Editing was detected by a sensitive and linear primer extension assay. An upstream region (nt 6648 to 6661) which affected the efficiency of editing was identified. RNAs with mutations in this efficiency sequence were edited at 22 to 160% of wild-type levels. Point mutations in a downstream 11-nt mooring sequence (nt 6671 to 6681) abolished editing, confirming previous studies (R. R. Shah, T. J. Knott, J. E. Legros, N. Navaratnam, J. C. Greeve, and J. Scott, J. Biol. Chem. 266:16301-16304, 1991). The optimal distance between the editing site and the mooring sequence is 5 nt, but a C positioned 8 nt upstream is edited even when nt 6666 contains U. The efficiency and mooring sequences were inserted individually and together adjacent to a heterologous C in apo-B mRNA. The mooring sequence alone induced editing of the C at nt 6597 both in vitro and in transfected rat hepatoma cells. Editing at nt 6597 was specific, was independent of editing at nt 6666, and was stimulated to wild-type levels when the efficiency sequence was also inserted. Introduction of the mooring sequence into a heterologous mRNA, luciferase mRNA, induced editing of an upstream cytidine. Although UV cross-linking studies have previously shown that proteins of 60 to 66 kDa cross-link to apo-B mRNA, these proteins did not cross-link to the luciferase translocation mutants. PMID- 8246951 TI - Interaction of two sequence-specific single-stranded DNA-binding proteins with an essential region of the beta-casein gene promoter is regulated by lactogenic hormones. AB - Transcription of the beta-casein gene in mammary epithelial cells is regulated by the lactogenic hormones insulin, glucocorticoids, and prolactin. The DNA sequence elements in the promoter which confer the action of the hormones on the transcriptional machinery and the nuclear proteins binding to this region have been investigated. We found that 221 nucleotides of promoter sequence 5' of the RNA start site are sufficient to mediate the induction of a chloramphenicol acetyltransferase reporter gene in transfected HC11 mammary epithelial cells. Deletion of 5' sequences to position -183 results in a construct with enhanced basal activity which still retains inducibility. A -170 beta-casein promoter chloramphenicol acetyltransferase construct has very low transcriptional activity, which indicates the presence of a negative regulatory in the region between -221 and -183 and a positive regulatory element between -183 and -170. Band shift analysis showed that the promoter region between -194 and -163 specifically binds two nuclear proteins. The proteins are sequence-specific, single-stranded DNA-binding proteins which exclusively recognize the upper DNA strand and most likely play a repressing role in transcription. DNA binding activity of these nuclear proteins was observed only in nuclear extracts from mammary glands of mice in late pregnancy and postlactation, not during lactation. Hormonal control of the DNA binding activity of these proteins was also observed in the mammary epithelial cell line HC11. Mixing experiments showed that extracts from mammary tissue of lactating mice and from lactogenic hormone-treated HC11 cells contain an activity which can suppress the DNA binding of the single stranded DNA-binding proteins.2+ identical specificity to the single-stranded DNA. PMID- 8246952 TI - Effector domain mutations dissociate p21ras effector function and GTPase activating protein interaction. AB - The GTPase activity of p21ras is stimulated by GTPase-activating proteins (GAPs) such as p120GAP and the product of the neurofibromatosis 1 gene, which may negatively regulate p21 function. GAPs are also proposed effectors of ras. We have sought activating substitutions in c-H-ras in the region encoding the effector domain, on the rationale that such mutations would dissociate effector function from negative regulation by GAP. One such activating mutation, Pro-34- >Arg, encodes protein that is substantially bound to GTP in vivo. In vitro, this protein is not stimulated by GAPs, and its binding to p120GAP is grossly impaired. The results support the idea that the p21 structural requirements for effector function and GAP interaction are quite different and suggest that some molecule(s) other than p120GAP serves as the ras effector. In contrast to the results obtained with p120GAP, the Pro-34-->Arg p21 species is effectively coupled to the raf-1 product, as judged from electrophoretic mobility shifts of the Raf-1 phosphoprotein. PMID- 8246953 TI - E2A expression, nuclear localization, and in vivo formation of DNA- and non-DNA binding species during B-cell development. AB - A monoclonal antibody (Yae) was characterized and shown to specifically recognize E2A proteins in vivo, including the E2A-Pbx1 fusion gene products, p77E2A-Pbx1 and p85E2A-Pbx1. E2A proteins of a predominant molecular mass of 72 kDa, which comigrated with in vitro-produced rat E12 and and rat E47, were detected in human pro-B, pre-B, mature B, and plasma cell lines. The Yae antibody detected an E2A containing microE2 enhancer element-binding complex (BCF-1) in pre-B- and mature B-cell lines in electrophoretic mobility shift assays which displayed a migration rate similar to that of in vitro-produced rat E12 and rat E47. A new E2A containing microE2-binding species (P-E2A) was identified in plasma cells by using electrophoretic mobility shift assays. E2A proteins were detected in pro-B cells but were unable to bind the microE2 site. These observations suggest that the microE2 site is the target of stage-specific E2A regulatory complexes during B-cell development. Immunostaining analyses demonstrated the predominant nuclear localization of E2A proteins. Finally, we have identified an E2A form, designated I-E2A, which is unable to bind DNA. Our observations demonstrate novel in vivo mechanisms for the regulation of transcription by E2A proteins during B-cell development. PMID- 8246954 TI - Oncogenic truncation of the first repeat of c-Myb decreases DNA binding in vitro and in vivo. AB - Oncogenic activation of c-Myb in both avian and murine systems often involves N terminal truncation. In particular, the first of three DNA-binding repeats in c Myb has been largely deleted during the genesis of the v-myb oncogenes of avian myeloblastosis virus and E26 avian leukemia virus. This finding suggests that the first DNA-binding repeat may have an important role in cell growth control. We demonstrate that truncation of the first DNA-binding repeat of c-Myb is sufficient for myeloid transformation in culture, but deletion of the N-terminal phosphorylation site and adjacent acidic region is not. Truncation of the first repeat decreases the ability of a Myb-VP16 fusion protein to trans activate the promoter of a Myb-inducible gene (mim-1) involved in differentiation. Moreover, truncation of the first repeat decreases the ability of the Myb protein to bind DNA both in vivo and in vitro. These results suggest that N-terminal mutants of c Myb may transform by regulating only a subset of those genes normally regulated by c-Myb. PMID- 8246955 TI - Unidirectional dominance of cytoplasmic inheritance in two genetic crosses of Plasmodium falciparum. AB - Malarial parasites have two highly conserved cytoplasmic DNA molecules: a 6-kb tandemly arrayed DNA that has characteristics of a mitochondrial genome, and a 35 kb circular DNA that encodes functions commonly found in chloroplasts. We examined the inheritance pattern of these elements in two genetic crosses of Plasmodium falciparum clones. Parent-specific oligonucleotide probes and single strand conformation polymorphism analysis identified single nucleotide changes that distinguished the parental 6- and 35-kb DNA molecules in the progeny. In all 16 independent recombinant progeny of a cross between a Central American clone, HB3, and a Southeast Asian clone, Dd2, the 6- and 35-kb DNAs were inherited from the Dd2 parent. In all nine independent recombinant progeny of a cross between clone HB3 and a likely African clone, 3D7, the 6-kb DNA was inherited from the 3D7 parent. Inheritance of cytoplasmic genomes of the Dd2 and 3D7 parents was, therefore, dominant over that of the HB3 parent. Cytoplasmic DNA molecules were found almost exclusively in the female gametes of malarial parasites; hence, clone HB3 did not appear to have served as a maternal parent for the progeny of two crosses. Defective differentiation into male gametes by clone Dd2 is likely to be a reason for the cytoplasmic inheritance pattern seen in the HB3 x Dd2 cross. However, incompetence of male or female gametes is unlikely to explain the uniparental dominance in recombinant progeny of the HB3 x 3D7 cross, since both parents readily self-fertilized and completed the malaria life cycle on their own. Instead, the data suggest unidirectional parental incompatibility in cross fertilization of these malarial parasites, where a usually cosexual parental clone can participate only as a male or as a female. Such an incompatibility may be speculated as indicating an early phase of reproductive isolation of P. falciparum clones from different geographical regions. PMID- 8246956 TI - Elevated levels of cyclin D1 protein in response to increased expression of eukaryotic initiation factor 4E. AB - Cyclin D1 is a G1-specific cyclin that has been linked to lymphoid, parathyroid, and breast tumors. Recent studies suggested that high protein levels of cyclin D1 are not always produced when cyclin D1 mRNA is overexpressed in transfected cells, suggesting that posttranscriptional events may be important in cyclin D1 regulation. The mRNA cap-binding protein (eukaryotic initiation factor 4E [eIF 4E]) is a potential regulatory of several posttranscriptional events, and it can itself induce neoplastic transformation. Consequently, we examined eIF-4E as a potential regulator of cyclin D1. Overexpression of cyclin D1 mRNA in NIH 3T3 cells did not increase cyclin D1 protein. In contrast, overexpression of eIF-4E markedly increased the amount of cyclin D1 protein in NIH 3T3 cells. This increase was specific to cyclin D1 in comparison with the retinoblastoma gene product, c-Myc, actin, and eukaryotic initiation factor 2 alpha. We also examined cyclin D1 protein in cells expressing an estrogen receptor-Myc fusion protein because we previously found that eIF-4E increases after induction of c-myc function. In these cells, increased levels of eIF-4E protein were closely followed by increases in levels of cyclin D1 protein, but the level of cyclin D1 mRNA was not increased. We conclude that increases in cyclin D1 levels may result from increased expression of eIF-4E, and this regulation may be one determinant of cyclin D1 levels in the cell. PMID- 8246957 TI - The essential yeast protein MIM44 (encoded by MPI1) is involved in an early step of preprotein translocation across the mitochondrial inner membrane. AB - The essential yeast gene MPI1 encodes a mitochondrial membrane protein that is possibly involved in protein import into the organelle (A. C. Maarse, J. Blom, L. A. Grivell, and M. Meijer, EMBO J. 11:3619-3628, 1992). For this report, we determined the submitochondrial location of the MPI1 gene product and investigated whether it plays a direct role in the translocation of preproteins. By fractionation of mitochondria, the mature protein of 44 kDa was localized to the mitochondrial inner membrane and therefore termed MIM44. Import of the precursor of MIM44 required a membrane potential across the inner membrane and involved proteolytic processing of the precursor. A preprotein in transit across the mitochondrial membranes was cross-linked to MIM44, whereas preproteins arrested on the mitochondrial surface or fully imported proteins were not cross linked. When preproteins were arrested at two distinct stages of translocation across the inner membrane, only preproteins at an early stage of translocation could be cross-linked to MIM44. Moreover, solubilized MIM44 was found to interact with in vitro-synthesized preproteins. We conclude that MIM44 is a component of the mitochondrial inner membrane import machinery and interacts with preproteins in an early step of translocation. PMID- 8246959 TI - The Ewing's sarcoma EWS/FLI-1 fusion gene encodes a more potent transcriptional activator and is a more powerful transforming gene than FLI-1. AB - EWS/FLI-1 is a chimeric protein formed by a tumor-specific 11;22 translocation found in both Ewing's sarcoma and primitive neuroectodermal tumor of childhood. EWS/FLI-1 has been shown to be a potent transforming gene, suggesting that it plays an important role in the genesis of these human tumors. We now demonstrate that EWS/FLI-1 has the characteristics of an aberrant transcription factor. Subcellular fractionation experiments localized the EWS/FLI-1 protein to the nucleus of primitive neuroectodermal tumor cells. EWS/FLI-1 specifically bound in vitro an ets-2 consensus sequence similarly to normal FLI-1. When coupled to a GAL4 DNA-binding domain, the amino-terminal EWS/FLI-1 region was a much more potent transcriptional activator than the corresponding amino-terminal domain of FLI-1. Finally, EWS/FLI-1 efficiently transformed NIH 3T3 cells, but FLI-1 did not. These data suggest that EWS/FLI-1, functioning as a transcription factor, leads to a phenotype dramatically different from that of cells expressing FLI-1. EWS/FLI-1 could disrupt normal growth and differentiation either by more efficiently activating FLI-1 target genes or by inappropriately modulating genes normally not responsive to FLI-1. PMID- 8246958 TI - Activation of the mouse mdr3 gene by insertion of retroviruses in multidrug resistant P388 tumor cells. AB - In multidrug-resistant (MDR) derivatives of the mouse lymphoid tumor P388, the emergence of MDR is associated with overexpression and transcriptional activation of the mdr3 gene, either in the absence of (P388/VCR-10) or concomitant with (P388/ADM-2) gene amplification. In both instances, Northern (RNA) blotting analyses have suggested the presence of altered mdr3 transcripts in these cells, possibly originating from novel transcription initiation sites. The mechanisms underlying mdr3 overexpression in these cells have been investigated. In P388/VCR 10 cells, Southern blotting analyses together with genomic DNA cloning and nucleotide sequencing have demonstrated the presence of an intact mouse mammary tumor virus (MMTV) within the boundaries of intron 1 of mdr3. cDNA cloning and nucleotide sequencing indicated that this integration event results in the synthesis and overexpression of a hybrid MMTV-mdr3 mRNA which initiates within the U3 region of the 5' long terminal repeat (LTR) of the provirus. Consequently, this mRNA lacks the normal exon 1 of mdr3 but contains (i) MMTV LTR-derived sequences at its 5' end, (ii) a novel mdr3 exon, mapping within the boundaries of intron 1 downstream of the MMTV integration site and generated by alternative splicing, and (iii) an otherwise intact 3' portion of mdr3 starting at exon 2. A similar type of analysis of P388/ADM-2 cells revealed that mdr3 overexpression in these cells is associated with the integration of an intracisternal A particle (IAP) within an L1Md repetitive element, immediately upstream of mdr3. The IAP insertion results in the overexpression of hybrid IAP-mdr3 mRNA transcripts that initiate within the 3' LTR of the IAP and which contain IAP LTR-derived sequences at the 5' end spliced 14 nucleotides upstream of the normal exon 1 of mdr3. Taken together, these results indicate that independent retroviral insertions were the initial mutagenic event responsible for mdr3 overexpression and survival during drug selection of these cell lines. Amplification of the rearranged and activated mdr3 gene copy occurred during further selection for high-level drug resistance in P388/ADM-2 cells. PMID- 8246960 TI - The granulocyte-macrophage colony-stimulating factor promoter cis-acting element CLE0 mediates induction signals in T cells and is recognized by factors related to AP1 and NFAT. AB - Expression of the granulocyte-macrophage colony-stimulating factor (GM-CSF) gene in T cells is activated by the combination of phorbol ester (phorbol myristate acetate) and calcium ionophore (A23187), which mimic antigen stimulation through the T-cell receptor. We have previously shown that a fragment containing bp -95 to +27 of the mouse GM-CSF promoter can confer inducibility to reporter genes in the human Jurkat T-cell line. Here we use an in vitro transcription system to demonstrate that a cis-acting element (positions -54 to -40), referred to as CLE0, is a target for the induction signals. We observed induction with templates containing intact CLE0 but not with templates with deleted or mutated CLE0. We also observed that two distinct signals were required for the stimulation through CLE0, since only extracts from cells treated with both phorbol myristate acetate and A23187 supported optimal induction. Stimulation probably was mediated by CLE0 binding proteins because depletion of these proteins specifically reduced GM-CSF transcription. One of the binding factors possessed biochemical and immunological features identical to those of the transcription factor AP1. Another factor resembled the T-cell-specific factor NFAT. The characteristics of these two factors are consistent with their involvement in GM-CSF induction. The presence of CLE0-like elements in the promoters of interleukin-3 (IL-3), IL-4, IL-5, GM CSF, and NFAT sites in the IL-2 promoter suggests that the factors we detected, or related factors that recognize these sites, may account for the coordinate induction of these genes during T-cell activation. PMID- 8246961 TI - Selective expression of intracisternal A-particle genes in established mouse plasmacytomas. AB - Mouse plasmacytomas generally express higher levels of RNA transcripts from endogenous intracisternal A-particle (IAP) proviral elements than do lipopolysaccharide-stimulated normal lymphocytes. Lymphocytes express a limited and highly characteristic set of IAP elements (lymphocyte-specific [LS] elements). In this study, we examined whether LS elements are expressed at higher levels after transformation of the cells and/or whether new IAP elements are activated. The IAP elements expressed in plasmacytoma MPC11 were characterized by sequence analysis of 22 cDNA clones. The long terminal repeats (LTRs) of the tumor cDNAs proved to be highly related in sequence. None of the clones was of the LS cDNA type. The MPC11 LTRs were five- to sixfold more active than an LS cDNA LTR when tested for promoter activity by transfection into plasmacytoma cells. The LTRs of the tumor-derived cDNAs contained a canonical ATF core sequence (ATF-PC), while the LS cDNAs contained an altered sequence (ATF-LS). An ATF-PC oligonucleotide probe detected multiple IAP transcripts on Northern (RNA) blots of RNA from several plasmacytoma but gave no reaction with RNA from stimulated B lymphocytes. In contrast, an ATF-LS probe detected higher levels of RNA in lymphocyte than in tumor RNAs. Thus, expression of IAP elements in transformed B cells is selective for a different set of regulatory sequence variants than those expressed in normal B cells. Other oligonucleotide probes representing LS- and PC-specific sequence variants detected multiple common hypomethylated IAP proviral loci in three independently derived plasmacytomas. Overall, the results show that established plasmacytomas exhibit a characteristic pattern of IAP proviral hypomethylation and regulatory sequence selection. PMID- 8246962 TI - Expression of trkA cDNA in neuroblastomas mediates differentiation in vitro and in vivo. AB - The human trkA cDNA was transfected into a malignant human neuroblastoma (NB) cell line (HTLA230) to investigate its role in NB growth and differentiation. This cell line lacks expression of both endogenous trkA and gp75NGFR genes. Transfectants expressing the trkA mRNA and surface-bound receptors transcriptionally activate immediate-early genes (c-fos, c-jun, and jun-B) following nerve growth factor (NGF) stimulation. NGF treatment induces growth arrest as well as down-regulation of the amplified N-myc oncogene. Genes selectively expressed in mature neurons (SCG-10, ret proto-oncogene, GAP-43, etc.) are transcriptionally activated, and neurite outgrowth further demonstrates differentiation of transfectants following NGF stimulation. trkA-expressing NB cells remain tumorigenic in nude mice; however, subcutaneous treatment of tumor bearing mice with NGF induces Schwannian and neuronal cell differentiation similar to the induction seen in human ganglioneuroblastomas. Thus, trkA expression in HTLA230 cells is sufficient to generate a functional NGF receptor complex that leads to growth-arrested and differentiated NB cells in vitro and in vivo in the presence of NGF. Hence, NGF may play a crucial role in NB cell differentiation and regression in vivo. PMID- 8246963 TI - Developmental regulation of the human embryonic beta-like globin gene is mediated by synergistic interactions among multiple tissue- and stage-specific elements. AB - The stage-specific regulation of mammalian embryonic globin genes has been an experimentally elusive problem, in part because of the developmentally early timing of their expression. We have carried out a systematic analysis of truncation and internal deletion mutations within the 5'-flanking region of the human embryonic beta-like globin gene (epsilon) in erythroid and nonerythroid cell lines. Within a 670-bp region upstream from the constitutive promoter are multiple positive and negative control elements. Of these, a positive regulatory element (epsilon-PRE II) which is active only in embryonic erythroid cells is of particular interest. Remarkably, although it is inactive on its own, in the presence of other sequences located further upstream, it confers tissue- and developmental stage-specific expression on a constitutive epsilon-globin or heterologous promoter. The activity of epsilon-PRE II is also modulated by another positive regulatory domain located further downstream to direct erythroid cell-specific, but little or no embryonic stage-specific, transcription. A nuclear factor highly enriched in embryonic erythroid cells binds specifically within a 19-bp region of epsilon-PRE II. Nuclei from adult erythroid cells also contain a factor that binds to this region but forms a complex of faster electrophoretic mobility. We speculate that interactions between epsilon-PRE II and other upstream control elements play an important role in the developmental regulation of the human embryonic beta-like globin gene. PMID- 8246964 TI - Properties of initiator-associated transcription mediated by GAL4-VP16. AB - Transcription associated with a terminal deoxynucleotide transferase gene initiator element is shown to respond to the transcription factor GAL4-VP16 both in vivo and in vitro. High-level transcription requires both an intact initiator element and bound activator. Transcription from this initiator-directed promoter is synergistic in vivo in that five GAL4 DNA binding sites yield 36 times the expression of a single site. Promoters dominated by initiator and TATA elements respond similarly to several GAL4-based activators, including GAL4-Sp1, GAL4-CTF, GAL4(1-147), GAL4-p53, GAL4-C/EBP, and GAL4-ER(EF), as well as GAL4-VP16 and Sp1. These and other similarities suggest that primary activation of TATA- and initiator-dominated promoters occurs at common steps. Since the initial assembly steps do not appear to be common for the two promoter types, the results place interesting constraints on models for how activation occurs. PMID- 8246965 TI - A novel hepatitis B virus (HBV) genetic element with Rev response element-like properties that is essential for expression of HBV gene products. AB - Many viruses possess complex mechanisms involving multiple gene products and cis regulatory elements in order to achieve a fine control of their gene expression at both transcriptional and posttranscriptional levels. Hepatitis B virus (HBV) and retroviruses share many structural and functional similarities. In this study, by genetic and biochemical analyses, we have demonstrated the existence of a novel genetic element within the HBV genome which is essential for high-level expression of viral gene products. This element is located 3' to the envelope coding region. We have shown that this genetic element is cis acting at the posttranscriptional level and that its function is exerted at the level of RNA processing as part of transcribed sequences. This RNA element is also functional in the context of a heterologous gene. Similar to the function of Rev-Rev response element interaction of human immunodeficiency virus type 1, this element appears to inhibit the splicing process and facilitate the transport and utilization of HBV transcripts. PMID- 8246966 TI - Yin-yang 1 activates the c-myc promoter. AB - Previous studies on the murine c-myc promoter demonstrated that a ubiquitously present protein, common factor 1 (CF1), bound at two sites located -260 and -390 bp from the P1 transcription start site. CF1 has been purified to near homogeneity and shown to be identical to the zinc finger protein Yin-yang 1 (YY1) as judged by similarity of molecular weight and other biochemical properties, immunological cross-reactivity, and the ability of recombinant YY1 to bind to CF1 sites. In cotransfection experiments, YY1 is a strong activator of transcription from c-myc promoter-based reporters. Furthermore, in murine erythroleukemia cells, overexpressed YY1 causes increased levels of c-myc mRNA initiated from both major transcription initiation sites of the endogenous c-myc gene. PMID- 8246967 TI - A position-dependent transcription-activating domain in TFIIIA. AB - Transcription of the Xenopus 5S RNA gene by RNA polymerase III requires the gene specific factor TFIIIA. To identify domains within TFIIIA that are essential for transcriptional activation, we have expressed C-terminal deletion, substitution, and insertion mutants of TFIIIA in bacteria as fusions with maltose-binding protein (MBP). The MBP-TFIIIA fusion protein specifically binds to the 5S RNA gene internal control region and complements transcription in a TFIIIA-depleted oocyte nuclear extract. Random, cassette-mediated mutagenesis of the carboxyl region of TFIIIA, which is not required for promoter binding, has defined a 14 amino-acid region that is critical for transcriptional activation. In contrast to activators of RNA polymerase II, the activity of the TFIIIA activation domain is strikingly sensitive to its position relative to the DNA-binding domain. When the eight amino acids that separate the transcription-activating domain from the last zinc finger are deleted, transcriptional activity is lost. Surprisingly, diverse amino acids can replace these eight amino acids with restoration of full transcriptional activity, suggesting that the length and not the sequence of this region is important. Insertion of amino acids between the zinc finger region and the transcription-activating domain causes a reduction in transcription proportional to the number of amino acids introduced. We propose that to function, the transcription-activating domain of TFIIIA must be correctly positioned at a minimum distance from the DNA-binding domain. PMID- 8246968 TI - Inactivation of c-Yes tyrosine kinase by elevation of intracellular calcium levels. AB - We have previously shown that the c-Src tyrosine kinase is activated four- to fivefold when cultured keratinocytes differentiate following the elevation of intracellular calcium levels. In contrast to c-Src, another Src family tyrosine kinase, c-Yes, was rapidly inactivated in these same cells, despite its marked similarity in structure and enzymatic activity to c-Src. The inactivation of c Yes was independent of the protein kinase C pathway, which is usually activated by elevation of intracellular calcium levels. The protein levels of c-Src and c Yes were not altered, but the phosphotyrosine content of both proteins was greatly reduced. As has been demonstrated for c-Src, in vitro dephosphorylation of c-Yes by incubation with protein tyrosine phosphatases also resulted in its activation, not inactivation. In vitro reconstitution experiments showed that c Yes can be inactivated by preincubation with a Ca(2+)-supplemented cell extract and that this inhibition was reversed by the addition of EGTA [ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid]. Gradient sedimentation of cell lysates showed that in cells treated with calcium and ionophore, c-Yes formed complexes with two distinct cellular proteins, whereas similar complexes were not seen in c-Src immunoprecipitates. One of these two proteins has the ability to inhibit c-Yes kinase activity in vitro. Finally, the Ca(2+)-dependent inactivation of c-Yes was observed in kidney tubular cells and fibroblasts, suggesting that the Ca(2+)-dependent regulation of c-Yes tyrosine kinase is not unique to keratinocytes. We postulate that c-Yes is inactivated through a Ca2+ dependent association with cellular proteins, which seems to override its activation resulting from tyrosine dephosphorylation. PMID- 8246969 TI - A nuclear tyrosine phosphatase downregulates interferon-induced gene expression. AB - Alpha and gamma interferons rapidly induce several early response genes in primary human diploid fibroblasts. The transcription rates of these genes are maximal after 1 h of interferon treatment and return to basal levels within 8 h. Three different interferon-activated DNA-binding complexes (ISGF3, GAF, and FcRF gamma) that are responsible for transcriptional activation of cellular genes have been characterized. Assembly of these complexes requires tyrosine phosphorylation of one or more of the protein components. In this report, we demonstrate that a nuclear tyrosine phosphatase is responsible for the deactivation of these interferon-regulated transcription factors and the subsequent transcriptional downregulation of the corresponding genes. Furthermore, tyrosine phosphorylation is required for nuclear localization of the 91-kDa protein that is part of all three interferon-induced transcription complexes. These results provide the first evidence for a nuclear tyrosine phosphatase activity as a mechanism of transcriptional regulation. PMID- 8246970 TI - The dynamics of chromatin condensation: redistribution of topoisomerase II in the 87A7 heat shock locus during induction and recovery. AB - We have examined the in vivo sites of action for topoisomerases II in the 87A7 heat shock locus as a function of gene activity. When the hsp70 genes are induced, there is a dramatic redistribution of topoisomerase II in the locus which parallels many of the observed alterations in chromatin structure. In addition to changes in the topoisomerase II distribution within the locus, we find topoisomerase II localized around the putative domain boundaries scs and scs'. During recovery, when the chromatin fiber of the locus recondenses, the major sites of action for topoisomerase II appear to be located within the two hsp70 genes and in the intergenic spacer separating the two genes. PMID- 8246971 TI - Interaction of the intron-encoded mobility endonuclease I-PpoI with its target site. AB - Endonucleases encoded by mobile group I introns are highly specific DNases that induce a double-strand break near the site to which the intron moves. I-PpoI from the acellular slime mold Physarum polycephalum mediates the mobility of intron 3 (Pp LSU 3) in the extrachromosomal nuclear ribosomal DNA of this organism. We showed previously that cleavage by I-PpoI creates a four-base staggered cut near the point of intron insertion. We have now characterized several further properties of the endonuclease. As determined by deletion analysis, the minimal target site recognized by I-PopI was a sequence of 13 to 15 bp spanning the cleavage site. The purified protein behaved as a globular dimer in sedimentation and gel filtration. In gel mobility shift assays in the presence of EDTA, I-PpoI formed a stable and specific complex with DNA, dissociating with a half-life of 45 min. By footprinting and interference assays with methidiumpropyl-EDTA iron(II), I-PpoI contacted a 22- to 24-bp stretch of DNA. The endonuclease protected most of the purines found in both the major and minor grooves of the DNA helix from modification by dimethyl sulfate (DMS). However, the reactivity to DMS was enhanced at some purines, suggesting that binding leads to a conformational change in the DNA. The pattern of DMS protection differed fundamentally in the two partially symmetrical halves of the recognition sequence. PMID- 8246972 TI - The Saccharomyces cerevisiae Cdc68 transcription activator is antagonized by San1, a protein implicated in transcriptional silencing. AB - The CDC68 gene (also called SPT16) encodes a transcription factor for the expression of a diverse set of genes in the budding yeast Saccharomyces cerevisiae. To identify other proteins that are functionally related to the Cdc68 protein, we searched for genetic suppressors of a cdc68 mutation. Four suppressor genes in which mutations reverse the temperature sensitivity imposed by the cdc68 1 mutation were found. We show here that one of the suppressor genes is the previously reported SAN1 gene; san1 mutations were originally identified as suppressors of a sir4 mutation, implicated in the chromatin-mediated transcriptional silencing of the two mating-type loci HML and HMR. Each san1 mutation, including a san1 null allele, reversed all aspects of the cdc68 mutant phenotype. Conversely, increased copy number of the wild-type SAN1 gene lowered the restrictive temperature for the cdc68-1 mutation. Our findings suggest that the San1 protein antagonizes the transcriptional activator function of the Cdc68 protein. The identification of san1 mutations as suppressors of cdc68 mutations suggests a role for Cdc68 in chromatin structure. PMID- 8246973 TI - Gal80 proteins of Kluyveromyces lactis and Saccharomyces cerevisiae are highly conserved but contribute differently to glucose repression of the galactose regulon. AB - We cloned the GAL80 gene encoding the negative regulator of the transcriptional activator Gal4 (Lac9) from the yeast Kluyveromyces lactis. The deduced amino acid sequence of K. lactis GAL80 revealed a strong structural conservation between K. lactis Gal80 and the homologous Saccharomyces cerevisiae protein, with an overall identity of 60% and two conserved blocks with over 80% identical residues. K. lactis gal80 disruption mutants show constitutive expression of the lactose/galactose metabolic genes, confirming that K. lactis Gal80 functions in essentially in the same way as does S. cerevisiae Gal80, blocking activation by the transcriptional activator Lac9 (K. lactis Gal4) in the absence of an inducing sugar. However, in contrast to S. cerevisiae, in which Gal4-dependent activation is strongly inhibited by glucose even in a gal80 mutant, glucose repressibility is almost completely lost in gal80 mutants of K. lactis. Indirect evidence suggests that this difference in phenotype is due to a higher activator concentration in K. lactis which is able to overcome glucose repression. Expression of the K. lactis GAL80 gene is controlled by Lac9. Two high-affinity binding sites in the GAL80 promoter mediate a 70-fold induction by galactose and hence negative autoregulation by Gal80. Gal80 in turn not only controls Lac9 activity but also has a moderate influence on its rate of synthesis. Thus, a feedback control mechanism exists between the positive and negative regulators. By mutating the Lac9 binding sites of the GAL80 promoter, we could show that induction of GAL80 is required to prevent activation of the lactose/galactose regulon in glycerol or glucose plus galactose, whereas the noninduced level of Gal80 is sufficient to completely block Lac9 function in glucose. PMID- 8246974 TI - Hematopoietic cell phosphatase associates with the interleukin-3 (IL-3) receptor beta chain and down-regulates IL-3-induced tyrosine phosphorylation and mitogenesis. AB - Hematopoietic cell phosphatase (HCP) is a tyrosine phosphatase with two Src homology 2 (SH2) domains that is predominantly expressed in hematopoietic cells, including cells whose growth is regulated by interleukin-3 (IL-3). The potential effects of HCP on IL-3-induced protein tyrosine phosphorylation and growth regulation were examined to assess the role of HCP in hematopoiesis. Our studies demonstrate that, following ligand binding, HCP specifically associates with the beta chain of the IL-3 receptor through the amino-terminal SH2 domain of HCP, both in vivo and in vitro, and can dephosphorylate the receptor chain in vitro. The effects of increasing or decreasing HCP levels in IL-3-dependent cells were assessed with dexamethasone-inducible constructs containing an HCP cDNA in sense and antisense orientations. Increased HCP levels were found to reduce the levels of IL-3-induced tyrosine phosphorylation of the receptor and to dramatically suppress cell growth. Conversely, decreasing the levels of HCP increased IL-3 induced tyrosine phosphorylation of the receptor and marginally increased growth rate. These results support a role for HCP in the regulation of hematopoietic cell growth and begin to provide a mechanistic explanation for the dramatic effects that the genetic loss of HCP, which occurs in motheaten (me) and viable motheaten (mev) mice, has on hematopoiesis. PMID- 8246975 TI - A coiled-coil oligomerization domain of Bcr is essential for the transforming function of Bcr-Abl oncoproteins. AB - In Philadelphia chromosome-positive human leukemias, the c-abl proto-oncogene on chromosome 9 becomes fused to the bcr gene on chromosome 22, and chimeric Bcr-Abl proteins are produced. The fused Bcr sequences activate the tyrosine kinase, actin-binding, and transforming functions of Abl. Activation of the Abl transforming function has been shown to require two distinct domains of Bcr: domain 1 (Bcr amino acids 1 to 63) and domain 2 (Bcr amino acids 176 to 242). The amino acid sequence of domain 1 indicates that it may be a coiled-coil oligomerization domain. We show here that domain 1 of Bcr forms a homotetramer. Tetramerization of Bcr-Abl through Bcr domain 1 correlates with activation of the tyrosine kinase and F-actin-binding functions of Abl. Disruption of the coiled coil by insertional mutagenesis inactivates the oligomerization function as well as the ability of Bcr-Abl to transform Rat-1 fibroblasts or to abrogate interleukin-3 dependence in lymphoid cells. These results strongly suggest that Bcr-Abl oligomers are the active entities in transformation. PMID- 8246976 TI - Signals in chicken beta-globin DNA influence chromatin assembly in vitro. AB - We have confirmed the result that chicken beta-globin gene chromatin, which possesses the characteristics of active chromatin in erythroid cells, has shortened internucleosome spacings compared with bulk chromatin or that of the ovalbumin gene, which is inactive. To understand how the short (approximately 180 bp) nucleosome repeat arises specifically on beta-globin DNA, we have studied chromatin assembly of cloned chicken beta-globin DNA in a defined in vitro system. With chicken erythrocyte core histones and linker histone H5 as the only cellular components, a cloned 6.2-kb chicken beta-globin DNA fragment assembled into chromatin possessing a regular 180 +/- 5-bp repeat, very similar to what is observed in erythroid cells. A 2-kb DNA subfragment containing the beta A gene and promoter region, but lacking the downstream intergenic region between the beta A and epsilon genes, failed to generate a regular nucleosome array in vitro, suggesting that the intergenic region facilitates linker histone-induced nucleosome alignment. When the beta A gene was placed on a plasmid that contained a known chromatin-organizing signal, nucleosome alignment with a 180-bp periodicity was restored, whereas nucleosomes on flanking plasmid sequences possessed a 210-bp spacing periodicity. Our results suggest that the shortened 180-bp nucleosome spacing periodicity observed in erythroid cells is encoded in the beta-globin DNA sequence and that nucleosome alignment by linker histones is facilitated by sequences in the beta A-epsilon intergenic region. PMID- 8246977 TI - A regulatory element in the CHA1 promoter which confers inducibility by serine and threonine on Saccharomyces cerevisiae genes. AB - CHA1 of Saccharomyces cerevisiae is the gene for the catabolic L-serine (L threonine) dehydratase, which is responsible for biodegradation of serine and threonine. We have previously shown that expression of the CHA1 gene is transcriptionally induced by serine and threonine. Northern (RNA) analysis showed that the additional presence of good nitrogen sources affects induction. This may well be due to inducer exclusion. To identify interactions of cis-acting elements with trans activators of the CHA1 promoter, we performed band shift assays of nuclear protein extracts with CHA1 promoter fragments. By this approach, we identified a protein-binding site of the CHA1 promoter. The footprint of this protein contains the ABF1-binding site consensus sequence. This in vitro binding activity is present irrespectively of CHA1 induction. By deletion analysis, two other elements of the CHA1 promoter, UAS1CHA and UAS2CHA, which are needed for induction of the CHA1 gene were identified. Each of the two sequence elements is sufficient to confer serine and threonine induction upon the CYC1 promoter when substituting its upstream activating sequence. Further, in a cha4 mutant strain which is unable to grow with serine or threonine as the sole nitrogen source, the function of UAS1CHA, as well as that of UAS2CHA, is obstructed. PMID- 8246978 TI - CTCF, a conserved nuclear factor required for optimal transcriptional activity of the chicken c-myc gene, is an 11-Zn-finger protein differentially expressed in multiple forms. AB - A novel sequence-specific DNA-binding protein, CTCF, which interacts with the chicken c-myc gene promoter, has been identified and partially characterized (V. V. Lobanenkov, R. H. Nicolas, V. V. Adler, H. Paterson, E. M. Klenova, A. V. Polotskaja, and G. H. Goodwin, Oncogene 5:1743-1753, 1990). In order to test directly whether binding of CTCF to one specific DNA region of the c-myc promoter is important for chicken c-myc transcription, we have determined which nucleotides within this GC-rich region are responsible for recognition of overlapping sites by CTCF and Sp1-like proteins. Using missing-contact analysis of all four nucleotides in both DNA strands and homogeneous CTCF protein purified by sequence-specific chromatography, we have identified three sets of nucleotides which contact either CTCF or two Sp1-like proteins binding within the same DNA region. Specific mutations of 3 of 15 purines required for CTCF binding were designed to eliminate binding of CTCF without altering the binding of other proteins. Electrophoretic mobility shift assay of nuclear extracts showed that the mutant DNA sequence did not bind CTCF but did bind two Sp1-like proteins. When introduced into a 3.3-kbp-long 5'-flanking noncoding c-myc sequence fused to a reporter CAT gene, the same mutation of the CTCF binding site resulted in 10- and 3-fold reductions, respectively, of transcription in two different (erythroid and myeloid) stably transfected chicken cell lines. Isolation and analysis of the CTCF cDNA encoding an 82-kDa form of CTCF protein shows that DNA-binding domain of CTCF is composed of 11 Zn fingers: 10 are of C2H2 class, and 1 is of C2HC class. CTCF was found to be abundant and conserved in cells of vertebrate species. We detected six major nuclear forms of CTCF protein differentially expressed in different chicken cell lines and tissues. We conclude that isoforms of 11-Zn-finger factor CTCF which are present in chicken hematopoietic HD3 and BM2 cells can act as a positive regulator of the chicken c-myc gene transcription. Possible functions of other CTCF forms are discussed. PMID- 8246979 TI - A novel 205-kilodalton testis-specific serine/threonine protein kinase associated with microtubules of the spermatid manchette. AB - To identify proteins which interact with and potentially modulate the function of microtubules during spermatogenesis, we prepared a total testis MAP (microtubule associated protein) antiserum and used it to isolate cDNA clones from a mouse testis cDNA expression library. Antibodies affinity purified by using one expression clone recognized a 205-kDa protein, termed MAST205, which colocalizes with the spermatid manchette. Sequencing of full-length cDNA clones encoding MAST205 revealed it to be a novel serine/threonine kinase with a catalytic domain related to those of the A and C families. The testis-specific MAST205 RNA increases in abundance during prepuberal testis development, peaking at the spermatid stage. The microtubule-binding region of MAST205 occupies a central region of the molecule including the kinase domain and sequences C terminal to this domain. Binding of MAST205 to microtubules requires interaction with other MAPs, since it does not bind to MAP-free tubulin. A 75-kDa protein associated with immunoprecipitates of MAST205 from extracts of both whole testis and testis microtubules becomes phosphorylated in in vitro kinase assays. This 75-kDa substrate of the MAST205 kinase may form part of the MAST205 protein complex which binds microtubules. The MAST205 protein complex may function to link the signal transduction pathway with the organization of manchette microtubules. PMID- 8246980 TI - Developmental regulation of human gamma-globin genes in transgenic mice. AB - We report results showing that several gamma gene promoter elements participate in the developmental control of gamma-globin genes. Four gamma gene constructs with 5' truncated at -141, -201, -382, and -730 of the A gamma gene promoter linked to a micro locus control region (microLCR) cassette were used for production of transgenic mice and analysis of gamma gene expression during development. Mice carrying a microLCR -141 A gamma construct displayed downregulation of gamma gene expression in the adult stage of development, indicating that the proximal promoter contains elements participating in gamma gene silencing. Mice carrying a microLCR -201 A gamma or a microLCR -382 A gamma construct displayed high gamma gene expression in the fetal stage of development and complete loss of gamma gene downregulation in the adult stage, suggesting that the -141 to -201 gamma gene sequence contains elements which upregulate gamma gene expression and are dominant over the negative element 3' to -141. Extension of the promoter to -730 resulted in reappearance of gamma gene downregulation, suggesting that the -382 to -730 sequences contain an adult-stage specific silencer. gamma gene expression in the microLCR -201 A gamma and the microLCR -382 A gamma transgenic mice was copy number dependent. All the microLCR -730 A gamma transgenic mice expressed gamma mRNA; however, gamma gene expression was copy number independent, indicating that levels of gamma gene expression were modulated by the surrounding chromatin. Our results suggest that multiple elements participate in gamma gene silencing. The findings in the microLCR-201 A gamma and microLCR -382 A gamma transgenic mice are interpreted to indicate that the LCR interacts not only with the minimal gamma gene promoter but also with sequences of the upstream promoter. We postulate that gamma gene downregulation is achieved when the interaction between LCR and the upstream promoter is disturbed by the silencer located in the -382 to -730 region. We propose that gamma gene silencing is achieved by the combined effect of negative elements located 3' to -141, the negative element located between -382 and -730, and the competition by the beta gene promoter during the adult stage of development. PMID- 8246981 TI - Hydrolysis of phosphatidylcholine couples Ras to activation of Raf protein kinase during mitogenic signal transduction. AB - We have investigated the relationship between hydrolysis of phosphatidylcholine (PC) and activation of the Raf-1 protein kinase in Ras-mediated transduction of mitogenic signals. As previously reported, cotransfection of a PC-specific phospholipase C (PC-PLC) expression plasmid bypassed the block to cell proliferation resulting from expression of the dominant inhibitory mutant Ras N 17. In contrast, PC-PLC failed to bypass the inhibitory effect of dominant negative Raf mutants, suggesting that PC-PLC functions downstream of Ras but upstream of Raf. Consistent with this hypothesis, treatment of quiescent cells with exogenous PC-PLC induced Raf activation, even when normal Ras function was blocked by Ras N-17 expression. Further, activation of Raf in response to mitogenic growth factors was blocked by inhibition of endogenous PC-PLC. Taken together, these results indicate that hydrolysis of PC mediates Raf activation in response to mitogenic growth factors. PMID- 8246982 TI - Purification, characterization, and cDNA cloning of an AU-rich element RNA binding protein, AUF1. AB - The degradation of some proto-oncogene and lymphokine mRNAs is controlled in part by an AU-rich element (ARE) in the 3' untranslated region. It was shown previously (G. Brewer, Mol. Cell. Biol. 11:2460-2466, 1991) that two polypeptides (37 and 40 kDa) copurified with fractions of a 130,000 x g postribosomal supernatant (S130) from K562 cells that selectively accelerated degradation of c myc mRNA in a cell-free decay system. These polypeptides bound specifically to the c-myc and granulocyte-macrophage colony-stimulating factor 3' UTRs, suggesting they are in part responsible for selective mRNA degradation. In the present work, we have purified the RNA-binding component of this mRNA degradation activity, which we refer to as AUF1. Using antisera specific for these polypeptides, we demonstrate that the 37- and 40-kDa polypeptides are immunologically cross-reactive and that both polypeptides are phosphorylated and can be found in a complex(s) with other polypeptides. Immunologically related polypeptides are found in both the nucleus and the cytoplasm. The antibodies were also used to clone a cDNA for the 37-kDa polypeptide. This cDNA contains an open reading frame predicted to produce a protein with several features, including two RNA recognition motifs and domains that potentially mediate protein-protein interactions. These results provide further support for a role of this protein in mediating ARE-directed mRNA degradation. PMID- 8246983 TI - A C/EBP-binding site in the transferrin promoter is essential for expression in the liver but not the brain of transgenic mice. AB - The gene for the iron-binding protein transferrin is transcribed at a high level in liver hepatocytes but is also active in several other cell types, including oligodendrocytes in the brain. Enhancer elements between bp -560 and -44 of the transferrin gene promoter specifically activated transcription from a heterologous promoter in transgenic mouse liver and brain. Within this region, a potent cis-acting element between bp -98 and -83 was found to be essential for gene activity in both cultured hepatocytes and transgenic mouse liver. The -98 to -83 element contains a CCAAT sequence and is specifically bound by a nuclear factor from mouse liver that is homologous to rat liver C/EBP (CAAT enhancer binding protein). Point mutations within this binding site inhibit factor binding and abolish transcription in transfected hepatoma cells. When placed in the context of the 3,000-bp transferrin promoter, the C/EBP binding site mutation causes a complete loss of transcription in transgenic mouse liver; however, transgene expression in the brain of the same animals was unaffected. These results suggest a modular structure for the transferrin promoter and demonstrate that deletions or specific point mutations can be used to generate transgene promoters with an activity more restricted than that of their endogenous counterparts. PMID- 8246984 TI - Cloning of a novel, ubiquitously expressed human phosphatidylinositol 3-kinase and identification of its binding site on p85. AB - Phosphatidylinositol 3-kinase (PI 3-kinase) has been implicated as a participant in signaling pathways regulating cell growth by virtue of its activation in response to various mitogenic stimuli. Here we describe the cloning of a novel and ubiquitously expressed human PI 3-kinase. The 4.8-kb cDNA encodes a putative translation product of 1,070 amino acids which is 42% identical to bovine PI 3 kinase and 28% identical to Vps34, a Saccharomyces cerevisiae PI 3-kinase involved in vacuolar protein sorting. Human PI 3-kinase is also similar to Tor2, a yeast protein required for cell cycle progression. Northern (RNA) analysis demonstrated expression of human PI 3-kinase in all tissues and cell lines tested. Protein synthesized from an epitope-tagged cDNA had intrinsic PI 3-kinase activity and associated with the adaptor 85-kDa subunit of PI 3-kinase (p85) in intact cells, as did endogenous human PI 3-kinase. Coprecipitation assays showed that a 187-amino-acid domain between the two src homology 2 domains of p85 mediates interaction with PI 3-kinase in vitro and in intact cells. These results demonstrate the existence of different PI 3-kinase isoforms and define a family of genes encoding distinct PI 3-kinase catalytic subunits that can associate with p85. PMID- 8246985 TI - AU-rich intronic elements affect pre-mRNA 5' splice site selection in Drosophila melanogaster. AB - cis-spliced nuclear pre-mRNA introns found in a variety of organisms, including Tetrahymena thermophila, Drosophila melanogaster, Caenorhabditis elegans, and plants, are significantly richer in adenosine and uridine residues than their flanking exons are. The functional significance of this intronic AU richness, however, has been demonstrated only in plant nuclei. In these nuclei, 5' and 3' splice sites are selected in part by their positions relative to AU-rich elements spread throughout the length of an intron. Because of this position-dependent selection scheme, a 5' splice site at the normal (+1) exon-intron boundary having only three contiguous consensus nucleotides can compete effectively with an enhanced exonic site (-57E) having nine consensus nucleotides and outcompete an enhanced site (+106E) embedded within the AU-rich intron. To determine whether transitions from AU-poor exonic sequences to AU-rich intronic sequences influence 5' splice site selection in other organisms, alleles of the pea rbcS3A1 intron were expressed in Drosophila Schneider 2 cells, and their splicing patterns were compared with those in tobacco nuclei. We demonstrate that this heterologous transcript can be accurately spliced in transfected Drosophila nuclei and that a +1 G-to-A knockout mutation at the normal splice site activates the same three cryptic 5' splice sites as in tobacco. Enhancement of the exonic (-57) and intronic (+106) sites to consensus splice sites indicates that potential splice sites located in the upstream exon or at the 5' exon-intron boundary are preferred in Drosophila cells over those embedded within AU-rich intronic sequences. In contrast to tobacco, in which the activities of two competing 5' splice sites upstream of the AU-rich intron are modulated by their proximity to the AU transition point, D. melanogaster utilizes the upstream site which has a higher proportion of consensus nucleotides. The enhanced version of the cryptic intronic site is efficiently selected in D. melanogaster when the normal +1 site is weakened or discrete AU-rich elements upstream of the +106E site are disrupted. Selection of this internal site in tobacco requires more drastic disruption of these motifs. We conclude that 5' splice site selection in Drosophila nuclei is influenced by the intrinsic strengths of competing sites and by the presence of AU-rich intronic elements but to a different extent than in tobacco. PMID- 8246986 TI - Formation of retinoid X receptor homodimers leads to repression of T3 response: hormonal cross talk by ligand-induced squelching. AB - Thyroid hormone receptors (TRs) form heterodimers with retinoid X receptors (RXRs). Heterodimerization is required for efficient TR DNA binding to most response elements and transcriptional activation by thyroid hormone. RXRs also function as auxiliary proteins for several other receptors. In addition, RXR alpha can be induced by specific ligands to form homodimers. Here we report that RXR-specific retinoids that induce RXR homodimers are effective repressors of the T3 response. We provide evidence that this repression by RXR-specific ligands occurs by sequestering of RXR from TR-RXR heterodimers into RXR homodimers. This ligand-induced squelching may represent an important mechanism by which RXR specific retinoids and 9-cis retinoic acid mediate hormonal cross talk among a subfamily of nuclear receptors activated by structurally unrelated ligands. PMID- 8246987 TI - Phosphatidylinositol (PI) 3-kinase and PI 4-kinase binding to the CD4-p56lck complex: the p56lck SH3 domain binds to PI 3-kinase but not PI 4-kinase. AB - CD4 serves as a receptor for major histocompatibility complex class II antigens and as a receptor for the human immunodeficiency virus type 1 (HIV-1) viral coat protein gp120. It is coupled to the protein-tyrosine kinase p56lck, an interaction necessary for an optimal response of certain T cells to antigen. In addition to the protein-tyrosine kinase domain, p56lck possesses Src homology 2 and 3 (SH2 and SH3) domains as well as a unique N-terminal region. The mechanism by which p56lck generates intracellular signals is unclear, although it has the potential to interact with various downstream molecules. One such downstream target is the lipid kinase phosphatidylinositol 3-kinase (PI 3-kinase), which has been found to bind to activated pp60src and receptor-tyrosine kinases. In this study, we verified that PI 3-kinase associates with the CD4:p56lck complex as judged by the presence of PI 3-phosphate generated from anti-CD4 immunoprecipitates and detected by high-pressure liquid chromatographic analysis. However, surprisingly, CD4-p56lck was also found to associate with another lipid kinase, phosphatidylinositol 4-kinase (PI 4-kinase). The level of associated PI 4 kinase was generally higher than PI 3-kinase activity. HIV-1 gp120 and antibody mediated cross-linking induced a 5- to 10-fold increase in the level of CD4 associated PI 4- and PI 3-kinases. The use of glutathione S-transferase fusion proteins carrying Lck-SH2, Lck-SH3, and Lck-SH2/SH3 domains showed PI 3-kinase binding to the SH3 domain of p56lck, an interaction facilitated by the presence of an adjacent SH2 domain. PI 4-kinase bound to neither the SH2 nor the SH3 domain of p56lck. CD4-p56lck contributes PI 3- and PI 4-kinase to the activation process of T cells and may play a role in HIV-1-induced immune defects. PMID- 8246988 TI - Regulated and constitutive activity by CDC25Mm (GRF), a Ras-specific exchange factor. AB - Serum stimulates cells to increase their proportion of Ras protein in the active GTP-bound state. We have recently identified four types (I to IV) of apparently full-length cDNAs from a single mammalian gene, called CDC25Mm or GRF, which is homologous to the Ras-specific exchange factor CDC25 of S. cerevisiae. The largest cDNA (type IV) is brain specific, with the other three classes, although they have distinct 5' ends, essentially representing progressive N-terminal deletions of this cDNA. When placed in a retroviral expression vector, all four types of cDNAs induced morphologic transformation of NIH 3T3 cells and an increase in the basal level of GTP.Ras. Serum stimulation of these transformants lead to a further increase in GTP.Ras only in cells expressing the type IV cDNA. Each type of GRF protein was found in cytosolic and membrane fractions, and the protein in each fraction could stimulate guanine nucleotide release from GDP.Ras in vitro. When NIH 3T3 cells and cells expressing the type IV protein were transfected with two versions of a mutant ras gene, one encoding membrane associated Ras protein and the other encoding a cytosolic Ras protein, the basal levels of GTP bound to both forms of the mutant Ras protein were significantly higher in the cells expressing the type IV protein. However, serum increased the level of GTP bound to the membrane-associated mutant Ras protein in NIH 3T3 cells and in cells expressing the type IV protein but not in cells expressing the cytosolic version of the Ras protein. We conclude that each type of CDC25Mm induces cell transformation via the ability of its C terminus to stimulate guanine nucleotide exchange on Ras, the presence of N-terminal sequences is associated with a serum-dependent change in GTP.Ras, and the serum-dependent increase in GTP.Ras by exogenous CDC25Mm or by endogenous exchange factors probably requires membrane association of both Ras and the exchange factor. PMID- 8246989 TI - Functional characterization of the L-type pyruvate kinase gene glucose response complex. AB - L-type pyruvate kinase (L-PK) gene expression is modulated by hormonal and nutritional conditions. We have previously shown that the glucose/insulin response element (GlRE) of the L-PK gene is built around two noncanonical E boxes (element L4) that cooperate closely with a contiguous binding site (element L3). We present in this report the identification of proteins that interact with both elements. The L3 site binds hepatocyte nuclear factor 4 (HNF4)- and COUP/TF related proteins. In fibroblasts, the overexpression of HNF4 transactivates the L PK promoter. On the contrary, COUP/TF strongly inhibits the active promoter in hepatocytes. The L4 site binds the major late transcription factor (MLTF) in vitro and ex vivo; mutations that suppress this binding activity also inactivated the GlRE function. Mutations transforming one or two noncanonical E boxes of element L4 into consensus MLTF/USF binding sites strongly increase the affinity for MLTF/USF and do not impair the glucose responsiveness. However, merely the ability to bind MLTF/USF does not seem to be sufficient to confer a GlRE activity: those elements in which one E box has been destroyed and the other has been transformed into a consensus MLTF/USF sequence bind MLTF/USF efficiently but do not confer a high glucose responsiveness on the L-PK gene promoter. Consequently, the full activity of the L-PK GlRE seems to require the cooperation between two putative MLTF/USF binding sites located in the vicinity of an HNF4 binding site. PMID- 8246990 TI - Sex-lethal autoregulation requires multiple cis-acting elements upstream and downstream of the male exon and appears to depend largely on controlling the use of the male exon 5' splice site. AB - The on/off state of the binary switch gene Sex-lethal (Sxl), which controls somatic sexual development in Drosophila melanogaster, is regulated at the level of alternative splicing. In males, in which the gene is off, the default splicing machinery produces nonfunctional mRNAs; in females, in which the gene is on, the autoregulatory activity of the Sxl proteins directs the splicing machinery to produce functional mRNAs. We have used germ line transformation to analyze the mechanism of default and regulated splicing. Our results demonstrate that a blockage mechanism is employed in Sxl autoregulation. However, in contrast to transformer, in which Sxl appears to function by preventing the interaction of splicing factors with the default 3' splice site, a different strategy is used in autoregulation. (i) Multiple cis-acting elements, both upstream and downstream of the male exon, are required. (ii) These cis-acting elements are distant from the splice sites they regulate, suggesting that the Sxl protein cannot function in autoregulation by directly competing with splicing factors for interaction with the regulated splice sites. (iii) The 5' splice site of the male exon appears to be dominant in regulation while the 3' splice site plays a subordinate role. PMID- 8246991 TI - The Saccharomyces cerevisiae DNA repair gene RAD23 encodes a nuclear protein containing a ubiquitin-like domain required for biological function. AB - In eukaryotes, the posttranslational conjugation of ubiquitin to various cellular proteins marks them for degradation. Interestingly, several proteins have been reported to contain ubiquitin-like (ub-like) domains that are in fact specified by the DNA coding sequences of the proteins. The biological role of the ub-like domain in these proteins is not known; however, it has been proposed that this domain functions as a degradation signal rendering the proteins unstable. Here, we report that the product of the Saccharomyces cerevisiae RAD23 gene, which is involved in excision repair of UV-damaged DNA, bears a ub-like domain at its amino terminus. This finding has presented an opportunity to define the functional significance of this domain. We show that deletion of the ub-like domain impairs the DNA repair function of RAD23 and that this domain can be functionally substituted by the authentic ubiquitin sequence. Surprisingly, RAD23 is highly stable, and the studies reported herein indicate that its ub-like domain does not mediate protein degradation. Thus, in RAD23 at least, the ub-like domain affects protein function in a nonproteolytic manner. PMID- 8246992 TI - A trypanosomal CCHC-type zinc finger protein which binds the conserved universal sequence of kinetoplast DNA minicircles: isolation and analysis of the complete cDNA from Crithidia fasciculata. AB - Replication of the kinetoplast DNA minicircle light strand initiates at a highly conserved 12-nucleotide sequence, termed the universal minicircle sequence. A Crithidia fasciculata single-stranded DNA-binding protein interacts specifically with the guanine-rich heavy strand of this origin-associated sequence (Y. Tzfati, H. Abeliovich, I. Kapeller, and J. Shlomai, Proc. Natl. Acad. Sci. USA 89:6891 6895, 1992). Using the universal minicircle sequence heavy-strand probe to screen a C. fasciculata cDNA expression library, we have isolated two overlapping cDNA clones encoding the trypanosomatid universal minicircle sequence-binding protein. The complete cDNA sequence defines an open reading frame encoding a 116-amino acid polypeptide chain consisting of five repetitions of a CCHC zinc finger motif. A significant similarity is found between this universal minicircle sequence-binding protein and two other single-stranded DNA-binding proteins identified in humans and in Leishmania major. All three proteins bind specifically to single-stranded guanine-rich DNA ligands. Partial amino acid sequence of the endogenous protein, purified to homogeneity from C. fasciculata, was identical to that deduced from the cDNA nucleotide sequence. DNA-binding characteristics of the cDNA-encoded fusion protein expressed in bacteria were identical to those of the endogenous C. fasciculata protein. Hybridization analyses reveal that the gene encoding the minicircle origin-binding protein is nuclear and may occur in the C. fasciculata chromosome as a cluster of several structural genes. PMID- 8246993 TI - Distinct cytoplasmic regions of the human granulocyte colony-stimulating factor receptor involved in induction of proliferation and maturation. AB - The granulocyte colony-stimulating factor receptor (G-CSF-R) transduces signals important for the proliferation and maturation of myeloid progenitor cells. To identify functionally important regions in the cytoplasmic domain of the G-CSF-R, we compared the actions of the wild-type receptor, two mutants, and a natural splice variant in transfectants of the mouse pro-B cell line BAF3 and two myeloid cell lines, 32D and L-GM. A region of 55 amino acids adjacent to the transmembrane domain was found to be sufficient for generating a growth signal. The immediate downstream sequence of 30 amino acids substantially enhanced the growth signaling in the three cell lines. In contrast, the carboxy-terminal part of 98 amino acids strongly inhibited growth signaling in the two myeloid cell lines but not in BAF3 cells. Truncation of this region lead to an inability of the G-CSF-R to transduce maturation signals in L-GM cells. An alternative carboxy tail present in a splice variant of the G-CSF-R also inhibited growth signaling, notably in both the myeloid cells and BAF3 cells, but appeared not to be involved in maturation. PMID- 8246994 TI - Hypovirulence-associated traits induced by a mycovirus of Cryphonectria parasitica are mimicked by targeted inactivation of a host gene. AB - Expression of the Vir2 gene of Cryphonectria parasitica is down-regulated in strains of the fungus containing a double-stranded RNA genetic element that reduces fungal virulence (W. A. Powell and N. K. Van Alfen, Mol. Cell. Biol. 7:3688-3693, 1987). We have sequenced the Vir2 gene and characterized its structure; the mRNA contains a short open reading frame whose product has structural similarities to several fungal pheromones. A null mutant was constructed by homologous recombination to determine the function of the Vir2 gene and whether its disruption resulted in any of the altered phenotypes exhibited by many hypovirulent strains, such as reductions in virulence, pigmentation, and sporulation. The Vir2 null mutant (18dm) exhibited a wild-type phenotype with respect to gross colony morphology, growth rate, pigmentation, asexual spore viability, and virulence in apple fruit and chestnut trees. However, numbers of asexual fruiting bodies (pycnidia) and conidia were reduced significantly in comparison with the wild-type strain EP155/2. In sexual crosses of 18dm with a wild-type strain of the opposite mating type, perithecia (sexual fruiting bodies) developed but were barren. Deletion of the Vir2 gene results in a phenotype that mimics that of many double-stranded-RNA-containing hypovirulent strains; i.e., the null mutant exhibits significant reductions in asexual sporulation and pycinidum production as well as impaired sexual crossing ability. To our knowledge, this is the first report of the partial reproduction of a virus induced phenotype by deletion of a virus-perturbed host gene. PMID- 8246995 TI - Cloning and characterization of E2F-2, a novel protein with the biochemical properties of transcription factor E2F. AB - E2F is a mammalian transcription factor that appears to play an important role in cell cycle regulation. While at least two proteins (E2F-1 and DP-1) with E2F-like activity have been cloned, studies from several laboratories suggest that additional homologs may exist. A novel protein with E2F-like properties, designated E2F-2, was cloned by screening a HeLa cDNA library with a DNA probe derived from the DNA binding domain of E2F-1 (K. Helin, J. A. Lees, M. Vidal, N. Dyson, E. Harlow, and A. Fattaey, Cell 70:337-350, 1992). E2F-2 exhibits overall 46% amino acid identity to E2F-1. Both the sequence and the function of the DNA and retinoblastoma gene product binding domains of E2F-1 are conserved in E2F-2. The DNA binding activity of E2F-2 is dramatically enhanced by complementation with particular sodium dodecyl sulfate-polyacrylamide gel electrophoresis purified components of HeLa cell E2F, and anti-E2F-2 antibodies cross-react with components of purified HeLa cell E2F. These observations are consistent with a model in which E2F binds DNA as a heterodimer of two distinct proteins, and E2F-2 is functionally and immunologically related to one of these proteins. PMID- 8246996 TI - The retinoblastoma protein binds to a family of E2F transcription factors. AB - E2F is a transcription factor that helps regulate the expression of a number of genes that are important in cell proliferation. Recently, several laboratories have isolated a cDNA clone that encodes an E2F-like protein, known as E2F-1. Subsequent characterization of this protein showed that it had the properties of E2F, but it was difficult to account for all of the suggested E2F activities through the function of this one protein. Using low-stringency hybridization, we have isolated cDNA clones that encode two additional E2F-like proteins, called E2F-2 and E2F-3. The chromosomal locations of the genes for E2F-2 and E2F-3 were mapped to 1p36 and 6q22, respectfully, confirming their independence from E2F-1. However, the E2F-2 and E2F-3 proteins are closely related to E2F-1. Both E2F-2 and E2F-3 bound to wild-type but not mutant E2F recognition sites, and they bound specifically to the retinoblastoma protein in vivo. Finally, E2F-2 and E2F-3 were able to activate transcription of E2F-responsive genes in a manner that was dependent upon the presence of at least one functional E2F binding site. These observations suggest that the E2F activities described previously result from the combined action of a family of proteins. PMID- 8246997 TI - A novel NF-kappa B complex containing p65 homodimers: implications for transcriptional control at the level of subunit dimerization. AB - The predominant inducible form of the NF-kappa B transcription factor is a heteromeric complex containing two Rel-related DNA-binding subunits, termed p65 and p50. Prior transfection studies have shown that when these p65 and p50 subunits are expressed independently as stable homodimers, p65 stimulates kappa B directed transcription, whereas p50 functions as a kappa B-specific repressor. While authentic p50 homodimers (previously termed KBF1) have been detected in nuclear extracts from nontransfected cells, experimental evidence supporting the existence of p65 homodimers in vivo was lacking. We now provide direct biochemical evidence for the presence of an endogenous pool of inducible p65 homodimers in intact human T cells. As with the prototypical NF-kappa B p50-p65 heterodimer, this novel p65 homodimeric form of NF-kappa B is functionally sequestered in the cytoplasm but rapidly appears in the nuclear compartment following cellular stimulation. Site-directed mutagenesis studies indicate that the homodimerization function of p65 is dependent upon the presence of cysteine 216 and a conserved recognition motif for protein kinase A (RRPS; amino acids 273 to 276), both of which reside within a 91-amino-acid segment of the Rel homology domain that mediates self-association. In contrast, mutations at these two sites do not affect heterodimerization of p65 with p50 or its functional interaction with I kappa B alpha. These later findings indicate that neither homo- nor heterodimer formation is an absolute prerequisite for I kappa B alpha recognition of p65. Taken together with prior in vivo transcription studies, these results suggest that the biological activities of p65 and p50 homodimers are independently regulated, thereby providing an integrated and flexible control mechanism for the rapid activation and repression of NF-kappa B/Rel-directed gene expression. PMID- 8246998 TI - Signals that produce 3' termini in CYC1 mRNA of the yeast Saccharomyces cerevisiae. AB - The cyc1-512 mutant was previously shown to contain a 38-bp deletion, 8 nucleotides upstream from the major wild-type poly(A) site, in the CYC1 gene, which encodes iso-1-cytochrome c of the yeast Saccharomyces cerevisiae. This 38 bp deletion caused a 90% reduction in the CYC1 transcripts, which were heterogeneous in size, aberrantly long, and presumably labile (K. S. Zaret and F. Sherman, Cell 28:563-573, 1982). Site-directed mutagenesis in and adjacent to the 38-bp region was used to identify signals involved in the formation and positioning of CYC1 mRNA 3' ends. In addition, combinations of various putative 3' end-forming signals were introduced by in vitro mutagenesis into the 3' region of the cyc1-512 mutant. The combined results from both studies suggest that 3' end formation in yeast cells involves signals having the following three distinct but integrated elements acting in concert: (i) the upstream element, including sequences TATATA, TAG ... TATGTA, and TTTTTATA, which function by enhancing the efficiency of downstream elements; (ii) downstream elements, such as TTAAGAAC and AAGAA, which position the poly(A) site; and (iii) the actual site of polyadenylation, which often occurs after cytidine residues that are 3' to the so called downstream element. While the upstream element is required for efficient 3' end formation, alterations of the downstream element and poly(A) sites generally do not affect the efficiency of 3' end formation but appear to alter the positions of poly(A) sites. In addition, we have better defined the upstream elements by examining various derivatives of TATATA and TAG ... TATGTA, and we have examined the spatial requirements of the three elements by systematically introducing or deleting upstream and downstream elements and cytidine poly(A) sites. PMID- 8246999 TI - A mutant androgen receptor from patients with Reifenstein syndrome: identification of the function of a conserved alanine residue in the D box of steroid receptors. AB - Reifenstein syndrome is an eponymic term that describes partial androgen insensitive disorders. Androgen receptor isolated from five patients with this syndrome contains a specific mutation in the DNA binding domain of the receptor. This mutation converts an alanine to a threonine at position 596 next to the zinc catenation site at the second finger. The threonine 596 mutant receptor mediated normal androgen response at promoters with closely positioned multiple regulatory elements for the androgen receptor and other transcription factors. Promoters with single isolated androgen response elements were not transactivated by the mutant receptor. In in vitro receptor-DNA binding studies, interaction with DNA by the mutant receptor was achieved only in the presence of an anti-androgen receptor antibody. Exchanging alanine 596 in the wild-type androgen receptor with serine or valine produced mutants with properties indistinguishable from those of the naturally occurring threonine 596 mutant receptor. These results indicate that an alanine residue at position 596 contributes important structural and functional activities to the androgen receptor. In the androgen receptor from the patients with Reifenstein syndrome, in which this alanine is converted to a threonine, wild-type receptor properties can be restored by exchanging an additional threonine at position 602 to an alanine. An alanine residue at position 596 or 602 in the DNA binding domain of the androgen receptor is therefore important for the full function of this receptor. In all steroid receptors that bind the core sequence AGAACANNNTGTTCT, an alanine residue is also present at a position equivalent to alanine 596 in the androgen receptor. PMID- 8247000 TI - Molecular cloning, expression, and characterization of the Drosophila 85 kilodalton TFIID subunit. AB - Transcription initiation factor TFIID is a multimeric protein complex that plays a central role in mediating promoter responses to various activators and repressors. To further understand the role of the 85-kDa TFIID subunit (p85), we have cloned the corresponding cDNA with a probe based on an amino acid sequence of the purified protein. The recombinant p85 interacts directly with both the TATA box-binding subunit (TFIID tau or TBP) and the 110-kDa subunit (p110) of TFIID, suggesting that p85 may play a role in helping to anchor p110 within the TFIID complex and, with other studies, that TFIID assembly and function may involve a concerted series of subunit interactions. Interestingly, the carboxy terminus of p85 contains eight of the WD-40 repeats found originally in the beta subunit of G proteins and more recently in other transcriptional regulatory factors. However, truncated p85 lacking all the WD-40 repeats maintained interactions with both TFIID tau and p110. These observations leave open the possibility of a distinct function for the WD-40 repeats, possibly in transducing signals by interactions with transcriptional regulators and/or other components of the basic transcriptional machinery. PMID- 8247001 TI - Mutational analysis of yeast profilin. AB - We have mutated two regions within the yeast profilin gene in an effort to functionally dissect the roles of actin and phosphatidylinositol 4,5-bisphosphate (PIP2) binding in profilin function. A series of truncations was carried out at the C terminus of profilin, a region that has been implicated in actin binding. Removal of the last three amino acids nearly eliminated the ability of profilin to bind polyproline in vitro but had no dramatic in vivo effects. Thus, the extreme C terminus is implicated in polyproline binding, but the physiological relevance of this interaction is called into question. More extensive truncation, of up to eight amino acids, had in vivo effects of increasing severity and resulted in changes in conformation and expression level of the mutant profilins. However, the ability of these mutants to bind actin in vitro was not eliminated, suggesting that this region cannot be solely responsible for actin binding. We also mutagenized a region of profilin that we hypothesized might be involved in PIP2 binding. Alteration of basic amino acids in this region produced mutant profilins that functioned well in vivo. Many of these mutants, however, were unable to suppress the loss of adenylate cyclase-associated protein (Cap/Srv2p [A. Vojtek, B. Haarer, J. Field, J. Gerst, T. D. Pollard, S. S. Brown, and M. Wigler, Cell 66:497-505, 1991]), indicating that a defect could be demonstrated in vivo. In vitro assays demonstrated that the inability to suppress loss of Cap/Srv2p correlated with a defect in the interaction with actin, independently of whether PIP2 binding was reduced. Since our earlier studies of Acanthamoeba profilins suggested the importance of PIP2 binding for suppression, we conclude that both activities are implicated and that an interplay between PIP2 binding and actin binding may be important for profilin function. PMID- 8247002 TI - The loop region of the helix-loop-helix protein Id1 is critical for its dominant negative activity. AB - Id1, a helix-loop-helix (HLH) protein which lacks a DNA binding domain, has been shown to negatively regulate other members of the HLH family by direct protein protein interactions, both in vitro and in vivo. In this study, we report the results of site-directed mutagenesis experiments aimed at defining the regions of Id1 which are important for its activity. We have found that the HLH domain of Id1 is necessary and nearly sufficient for its activity. In addition, we show that two amino acid residues at the amino terminus of the Id1 loop are critical for its activity, perhaps by specifying the correct dimerization partners. In this regard, replacing the first four amino acids of the loops of the basic HLH proteins E12 and E47 with the corresponding amino acids of Id1 confers Id1 dimerization specificity. These studies point to the loop region as an important structural and functional element of the Id subfamily of HLH proteins. PMID- 8247003 TI - Identification of a cis-regulatory element and putative trans-acting factors responsible for 12-O-tetradecanoylphorbol-13-acetate (TPA)-mediated induction of heme oxygenase expression in myelomonocytic cell lines. AB - The human monocytic leukemia cell line THP-1 differentiates into macrophage-like cells when treated with a variety of agents, including 12-O-tetradecanoylphorbol 13-acetate (TPA). We show here that during this process, the expression of heme oxygenase, a rate-limiting enzyme in heme catabolism, is induced. Treatment with TPA increases heme oxygenase mRNA in other myelomonocytic cell lines also, but not in cell lines of other lineages, such as HeLa cells. Increased heme oxygenase activity may represent one of the functions of activated macrophages, which sequestrate senescent erythrocytes and degrade heme derived from hemoglobin. This cell-type-specific induction by TPA treatment further investigated with respect to transcriptional regulation. We defined a cis-regulatory element, 5'-GTCATATGAC 3', located in the 5'-flanking region (positions -156 to -147) of the human heme oxygenase gene, which confers inducibility by TPA in THP-1 cells but not in HeLa cells. Nuclear proteins that bind to this element, which may be responsible for the cell specificity, were identified in THP-1 nuclear extracts. This element contains the consensus motif CANNTG, to which a large family of basic helix-loop helix proteins binds. Our results suggest a novel mechanism of TPA-mediated transcriptional regulation in myelomonocytic cell lines. PMID- 8247004 TI - Identification and sequence analysis of cDNAs encoding a 110-kilodalton actin filament-associated pp60src substrate. AB - Transformation of chicken embryo cells by oncogenic forms of pp60src (e.g., pp60v src or pp60527F) is linked with a concomitant increase in the steady-state levels of tyrosine-phosphorylated cellular proteins. Activated forms of the Src protein tyrosine kinase stably associate with tyrosine-phosphorylated proteins, including a protein of 110 kDa, pp110. Previous reports have established that stable complex formation between pp110 and pp60src requires the structural integrity of the Src SH2 and SH3 domains, whereas tyrosine phosphorylation of pp110 requires only the structural integrity of the SH3 domain. In normal chicken embryo cells, pp110 colocalizes with actin stress filaments, and in Src-transformed cells, pp110 is found associated with podosomes (rosettes). Here, we report the identification and characterization of cDNAs encoding pp110. The predicted open reading frame encodes a polypeptide of 635 amino acids which exhibits little sequence similarity with other protein sequences present in the available sequence data bases. Thus, pp110 is a distinctive cytoskeleton-associated protein. On the basis of its association with actin stress filaments, we propose the term AFAP-110, for actin filament-associated protein of 110 kDa. In vitro analysis of AFAP-110 binding to bacterium-encoded glutathione S-transferase (GST) fusion proteins revealed that AFAP-110 present in normal cell extracts binds efficiently to Src SH3/SH2-containing fusion proteins, less efficiently to Src SH3-containing proteins, and poorly to SH2-containing fusion proteins. In contrast, AFAP-110 in Src-transformed cell extracts bound to GST-SH3/SH2 and GST SH2 fusion proteins. Analysis of AFAP-110 cDNA sequences revealed the presence of sequence motifs predicted to bind to SH2 and SH3 domains, respectively. We suggest that AFAP-110 may represent a cellular protein capable of interacting with SH3-containing proteins and, upon tyrosine phosphorylation, binds tightly to SH2-containing proteins, such as pp60src or pp59fyn. The potential roles of AFAP 110 as an SH3/SH2 cytoskeletal binding protein are discussed. PMID- 8247005 TI - DRS1 to DRS7, novel genes required for ribosome assembly and function in Saccharomyces cerevisiae. AB - To identify Saccharomyces cerevisiae mutants defective in assembly or function of ribosomes, a collection of cold-sensitive strains generated by treatment with ethyl methanesulfonate was screened by sucrose gradient analysis for altered ratios of free 40S to 60S ribosomal subunits or qualitative changes in polyribosome profiles. Mutations defining seven complementation groups deficient in ribosomal subunits, drs1 to drs7, were identified. We have previously shown that DRS1 encodes a putative ATP-dependent RNA helicase necessary for assembly of 60S ribosomal subunits (T. L. Ripmaster, G. P. Vaughn, and J. L. Woolford, Jr., Proc. Natl. Acad. Sci. USA 89:11131-11135, 1992). Strains bearing the drs2 mutation process the 20S precursor of the mature 18S rRNA slowly and are deficient in 40S ribosomal subunits. Cloning and sequencing of the DRS2 gene revealed that it encodes a protein similar to membrane-spanning Ca2+ ATPases. The predicted amino acid sequence encoded by DRS2 contains seven transmembrane domains, a phosphate-binding loop found in ATP- or GTP-binding proteins, and a seven-amino-acid sequence detected in all classes of P-type ATPases. The cold sensitive phenotype of drs2 is suppressed by extra copies of the TEF3 gene, which encodes a yeast homolog of eukaryotic translation elongation factor EF-1 gamma. Identification of gene products affecting ribosome assembly and function among the DNAs complementing the drs mutations validates the feasibility of this approach. PMID- 8247006 TI - The essential mitotic target of calmodulin is the 110-kilodalton component of the spindle pole body in Saccharomyces cerevisiae. AB - Two independent methods identified the spindle pole body component Nuf1p/Spc110p as the essential mitotic target of calmodulin. Extragenic suppressors of cmd1-1 were isolated and found to define three loci, XCM1, XCM2, and XCM3 (extragenic suppressor of cmd1-1). The gene encoding a dominant suppressor allele of XCM1 was cloned. On the basis of DNA sequence analysis, genetic cosegregation, and mutational analysis, XCM1 was identified as NUF1/SPC110. Independently, a C terminal portion of Nuf1p/Spc110p, amino acid residues 828 to 944, was isolated as a calmodulin-binding protein by the two-hybrid system. As assayed by the two hybrid system, Nuf1p/Spc110p interacts with wild-type calmodulin and triple mutant calmodulins defective in binding Ca2+ but not with two mutant calmodulins that confer a temperature-sensitive phenotype. Deletion analysis by the two hybrid system mapped the calmodulin-binding site of Nuf1p/Spc110p to amino acid residues 900 to 927. Direct binding between calmodulin and Nuf1p/Spc110p was demonstrated by a modified gel overlay assay. Furthermore, indirect immunofluorescence with fixation procedures known to aid visualization of spindle pole body components localized calmodulin to the spindle pole body. Sequence analysis of five suppressor alleles of NUF1/SPC110 indicated that suppression of cmd1-1 occurs by C-terminal truncation of Nuf1p/Spc110p at amino acid residues 856, 863, or 881, thereby removing the calmodulin-binding site. PMID- 8247007 TI - Premature termination of tubulin gene transcription in Xenopus oocytes is due to promoter-dependent disruption of elongation. AB - We have shown previously that the Xenopus alpha-tubulin gene, X alpha T14, exhibits premature termination of transcription when injected into oocyte nuclei. The 3' ends of prematurely terminated transcripts are formed immediately downstream of a stem-loop sequence found in the first 41 bp of the 5' leader. We show here, using deleted constructs, that premature termination requires the presence only of sequences from -200 to +19 relative to the initiation site. Deletion of the stem-loop does not increase the production of extended transcripts, and premature termination apparently continues at nonspecific sites. This finding indicates that disruption of the elongation phase of transcription rather than abrogation of a specific antitermination mechanism is the cause of premature termination in X alpha T14. We also found that disruption of elongation on a reporter gene could be induced specifically by competition with X alpha T14 promoters. To identify which elements of the promoter might interact with elongation determinants to cause this competition, we constructed a series of internal promoter mutants. Most mutations in the -200 to -60 region of the promoter had some effect on initiation frequency but did not cause any significant change in levels of premature termination. However, mutations in the core promoter that removed the TATA box consensus causes major change in initiation and resulted in a marked decrease in the production of prematurely terminated transcripts relative to extended transcripts. We discuss why such promoters can apparently escape the disruption of elongation that leads to premature termination. PMID- 8247008 TI - Nuclear RNase MRP is required for correct processing of pre-5.8S rRNA in Saccharomyces cerevisiae. AB - RNase MRP is a site-specific ribonucleoprotein endoribonuclease that cleaves RNA from the mitochondrial origin of replication in a manner consistent with a role in priming leading-strand DNA synthesis. Despite the fact that the only known RNA substrate for this enzyme is complementary to mitochondrial DNA, the majority of the RNase MRP activity in a cell is found in the nucleus. The recent characterization of this activity in Saccharomyces cerevisiae and subsequent cloning of the gene coding for the RNA subunit of the yeast enzyme have enabled a genetic approach to the identification of a nuclear role for this ribonuclease. Since the gene for the RNA component of RNase MRP, NME1, is essential in yeast cells and RNase MRP in mammalian cells appears to be localized to nucleoli within the nucleus, we utilized both regulated expression and temperature-conditional mutations of NME1 to assay for a possible effect on rRNA processing. Depletion of the RNA component of the enzyme was accomplished by using the glucose-repressed GAL1 promoter. Shortly after the shift to glucose, the RNA component of the enzyme was found to be depleted severely, and rRNA processing was found to be normal at all sites except the B1 processing site. The B1 site, at the 5' end of the mature 5.8S rRNA, is actually composed of two cleavage sites 7 nucleotides apart. This cleavage normally generates two species of 5.8S rRNA at a ratio of 10:1 (small to large) in most eukaryotes. After RNase MRP depletion, yeast cells were found to have almost exclusively the larger species of 5.8S rRNA. In addition, an aberrant 309-nucleotide precursor that stretched from the A2 to E processing sites of rRNA accumulated in these cells. Temperature-conditional mutations in the RNase MRP RNA gene gave an identical phenotype. Translation in yeast cells depleted of the smaller 5.8S rRNA was found to remain robust, suggesting a possible function for two 5.8S rRNAs in the regulated translation of select messages. These results are consistent with RNase MRP playing a role in a late step of rRNA processing. The data also indicate a requirement for having the smaller form of 5.8S rRNA, and they argue for processing at the B1 position being composed of two separate cleavage events catalyzed by two different activities. PMID- 8247009 TI - Complementation by wild-type p53 of interleukin-6 effects on M1 cells: induction of cell cycle exit and cooperativity with c-myc suppression. AB - Stable transfection of M1 myeloid leukemia cells with a temperature-sensitive mutant of p53 results in two phenomena that are manifested exclusively at the permissive temperature. On one hand, activation of wild-type p53 by the temperature shift induced an apoptotic type of cell death which could be inhibited by interleukin-6 (IL-6) (E. Yonish-Rouach, D. Resnitzky, J. Lotem, L. Sachs, A. Kimchi, and M. Oren, Nature 352:345-347, 1991). On the other hand, as reported in this work, activated p53 complemented the antiproliferative effects of IL-6 in M1 cells. A shift to the permissive temperature concomitant with or early after IL-6 treatment imposed a novel pattern of cell cycle arrest in which about 95% of the cells were retained within a G0-like quiescent state. This phase was characterized by 2N DNA content and low RNA and protein content. On the molecular level, activation of wild-type p53 transrepressed the c-myc gene but not the cyclin A, D1, or D2 gene, which are all independently suppressed by IL-6 in M1 cells. To further analyze whether c-myc inhibition mediates or complements p53 effects, the p53-transfected M1 cells were infected with a retroviral vector expressing deregulated c-myc, refractory to p53 or IL-6 action. It was found that the process of cell death was not interrupted at all in these M1 c-myc-p53 double transfectants, suggesting that the transrepression of c-myc is not a major obligatory event mediating p53-induced cell death. In addition, some of the antiproliferative effects of activated p53, manifested in the presence of IL-6, could still be transmitted in the background of constitutive c-myc. Yet the context of deregulated c-myc interfered with the final accumulation of cells within a G0-like phase, suggesting complementary interactions between the outcome of p53 activation and of c-myc suppression in the control of cell cycle arrest. PMID- 8247010 TI - TATA-binding protein and associated factors in polymerase II and polymerase III transcription. AB - Transcription by RNA polymerase I (pol I), pol II, and pol III requires the TATA binding protein (TBP). This protein functions in association with distinct TBP associated factors (TAFs) which may specify the nature of the polymerase selected for initiation at a promoter site. In the pol III transcription system, the TBP TAF complex is a component of the TFIIIB factor. This factor has been resolved into a TBP-TAF complex and another component, both of which are required for reconstitution of transcription by pol III. Neither the TBP-TAF complexes B-TFIID and D-TFIID, which were previously characterized as active for pol II transcription, nor TBP alone can complement pol III transcription reactions that are dependent upon the TBP-TAF subcomponent of TFIIIB. Surprisingly, the TBP-TAF subcomponent of TFIIIB is active in reconstitution of pol II transcription. PMID- 8247011 TI - The responsiveness of embryonic stem cells to alpha and beta interferons provides the basis of an inducible expression system for analysis of developmental control genes. AB - Embryonic stem (ES) cells, derived from the inner cell mass of the preimplantation mouse embryo, are used increasingly as an experimental tool for the investigation of early mammalian development. The differentiation of these cells in vitro can be used as an assay for factors that regulate early developmental decisions in the embryo, while the effects of altered gene expression during early embryogenesis can be analyzed in chimeric mice generated from modified ES cells. The experimental versatility of ES cells would be significantly increased by the development of systems which allow precise control of heterologous gene expression. In this paper, we report that ES cells are responsive to alpha and beta interferons (IFNs). This property has been exploited for the development of inducible ES cell expression vectors, using the promoter of the human IFN-inducible gene, 6-16. The properties of these vectors have been analyzed in both transiently and stably transfected ES cells. Expression was minimal or absent in unstimulated ES cells, could be stimulated up to 100-fold by treatment of the cells with IFN, and increased in linear fashion with increasing levels of IFN. High levels of induced expression were maintained for extended periods of time in the continuous presence of the inducing signal or following a 12-h pulse with IFN. Treatment of ES cells with IFN did not affect their growth or differentiation in vitro or compromise their developmental potential. This combination of features makes the 6-16-based expression vectors suitable for the functional analysis of developmental control control genes in ES cells. PMID- 8247012 TI - Decreased levels of internalized thyrotropin-releasing hormone receptors after uncoupling from guanine nucleotide-binding protein and phospholipase-C. AB - Internalization of TRH receptor (TRH-R) is dependent on sequences/structures in the receptor carboxyl-terminal tail. Here, we studied whether coupling to guanine nucleotide-binding protein (G-protein) and phospholipase-C (PLC) is involved in internalization. We constructed two mutant TRH-Rs: delta 218-263 TRH-R, in which most of the residues that form the putative third intracellular loop were deleted, and D71A TRH-R, in which an Asp in the putative second transmembrane helix was mutated to Ala; these TRH-Rs did not activate PLC when expressed transiently in COS-1 cells. In contrast to wild-type (WT) TRH-Rs, approximately 60% of which were internalized at steady state after binding methyl-HisTRH, only approximately 15% of delta 218-263 and D71A TRH-Rs were internalized. Thus, mutant TRH-Rs that do not activate PLC, most likely because they are uncoupled from G-proteins, are internalized to lesser extents than WT TRH-Rs. We also studied the effects of U73122 (1-[6-[[17 beta-3-methoxyestra-1,3,5(10)-trien-17 yl]amino] hexyl]-1H-pyrrole-2,5-dione), an amino steroid that inhibits receptor mediated activation of PLC. In COS-1 and AtT-20 cells transfected with WT TRH-Rs and in GH3 cells, U73122 virtually abolished TRH activation of PLC and partially reduced the fraction of WT TRH-Rs internalized. Thus, uncoupling WT TRH-Rs from PLC decreases internalization. We conclude that TRH-R coupling to G-protein and PLC increases the number of TRH-Rs internalized at steady state even though the primary signals for agonist-induced internalization are present in the receptor. These data support the idea that a quaternary complex of TRH/TRH-R/G protein/PLC is normally internalized. PMID- 8247013 TI - Dominant and nondominant negative C-erbA beta 1 receptors associated with thyroid hormone resistance syndromes augment 12-O-tetradecanoyl-phorbol-13-acetate induction of the collagenase promoter and exhibit defective 3,5,3' triiodothyronine-mediated repression. AB - C-erbA receptors and v-erbA have been shown to functionally interact with 12-O tetradecanoyl-phorbol-13-acetate (TPA)-inducible gene expression. These proteins enhance trans-activation by c-jun, and the c-erbA receptors in the presence of thyroid hormone repress TPA and c-jun induction of transcription. Also, v-erbA can abrogate T3-mediated repression. We have examined how dominant negative (S and CL) and nondominant negative (G-H) receptors cloned from various patients with thyroid hormone resistance syndromes affect expression of the collagenase promoter induced with TPA. The CL receptor (ARG315HIS mutation) has a 2-fold reduction in T3-binding affinity compared with human c-erbA beta 1 wild-type (WT) receptor, whereas the G-H receptor (ARG311HIS) and S receptor (deletion, THR codon 332) have T3-binding affinities reduced by 100-fold and greater than 100 fold, respectively. These mutant receptors were cotransfected with a collagenase promoter (-1200 to +63 base pairs) chloramphenicol acetyltransferase reporter gene (Col-CAT) into COS-7 cells. Levels of CAT reporter gene expression after transient transfection were determined in the presence or absence of 3-10 nM T3 and the presence or absence of 100 nM TPA. Unoccupied CL receptor and G-H and S receptors stimulated TPA-induced Col-CAT expression 1.5- to 9-fold. The CL receptor with thyroid hormone totally repressed TPA induction of the collagenase receptor. In the presence of thyroid hormone, the enhancing effects by S and G-H receptors on TPA-induced Col-CAT expression were unaffected and minimally diminished, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247014 TI - Glucocorticoids induce a G1/G0 cell cycle arrest of Con8 rat mammary tumor cells that is synchronously reversed by steroid withdrawal or addition of transforming growth factor-alpha. AB - Con8 mammary tumor cells are an epithelial cell line derived from the 7,12 dimethylbenz(alpha)anthracene-induced 13762NF rat mammary adenocarcinoma. The synthetic glucocorticoid dexamethasone suppresses the growth of Con8 cells, and after 5 days of treatment with this steroid, Con8 cells undergo less than 0.5 population doublings. This growth arrest is accompanied by a 30-fold elevation in c-jun transcript levels, no change in c-fos expression, and a moderate increase in total AP-1 transcriptional activity. Dexamethasone inhibited DNA synthesis within one cell cycle, and flow cytometry of propidium iodide-stained nuclei demonstrated that dexamethasone growth-suppressed cells had a DNA content indicative of a specific cell cycle block in either G1 or G0. Consistent with a G1/G0 arrest of the cell cycle, dexamethasone did not prevent Con8 cells from entering the S phase after release from synchronization at the G1/S boundary by a double thymidine block. Analysis of [3H]thymidine incorporation and autoradiography of [3H]thymidine-labeled nuclei revealed that after either dexamethasone withdrawal or the addition of transforming growth factor-alpha (TGF alpha), Con8 cells synchronously reinitiate cell cycle progression. Northern blot analysis demonstrated that an induction of transcripts for the G1 marker genes c myc and cyclin D1 occurs before cells enter the S-phase. After dexamethasone withdrawal, c-myc and cyclin D1 expression transiently peak at 2 and 4 h, respectively. In contrast, c-myc expression peaked at 0.5-1 h, whereas cyclin D1 expression was induced at 2 h and maintained at a high level after the addition of TGF alpha. Our results demonstrate that glucocorticoids induce a specific block of the cell cycle progression of a rat mammary tumor cell, and that after synchronous progression through the cell cycle, the temporal expression pattern for c-myc and cyclin D1 is distinct for dexamethasone release vs. the addition of TGF alpha to glucocorticoid-suppressed cells. PMID- 8247015 TI - Endogenous protein kinase-C activation in osteoblast-like cells modulates responsiveness to estrogen and estrogen receptor levels. AB - The osteoblast-like osteosarcoma cell line ROS 17/2.8, which expresses very low levels of estrogen receptor (ER), was stably transfected with the mouse ER in order to more easily evaluate the physiological role of estrogens in bone cell homeostasis. These transfected ROS.SMER 14 cells are highly responsive to estrogenic stimulation at subconfluence, but become refractory to estrogenic stimulation when postconfluency is reached. The purpose of these studies was to determine the mechanisms underlying this loss of responsiveness in these ER stably transfected cells at postconfluence. When proliferative capacity was evaluated by bromodeoxyuridine immunocytochemistry, approximately 70% of the subconfluent cells were actively dividing, whereas none of the postconfluent cells underwent division. Subconfluent cells were found to contain 2500-3000 ER binding sites/cell, whereas the ER in postconfluent cells was low and often undetectable. Steady state ER mRNA levels were not significantly modified by postconfluency. ER protein levels were also unaffected by confluency status. Since protein kinase-C (PKC) has been reported to influence cell proliferation and steroid hormone receptor binding, PKC activity was measured in sub- and postconfluent cells. Calcium-dependent PKC activity was approximately about 2 fold higher in postconfluent compared to subconfluent cells, whereas no differences were discerned in calcium-independent PKC activity. In an effort to examine the role of PKC in greater detail, postconfluent cells were treated with PKC inhibitors (H-7 or staurosporine) or with the tumor promoter TPA (12-O tetradecanoylphorbol-13-acetate) to down-regulate PKC activity, and changes in ER were evaluated. Inhibition or down-regulation of the PKC activity in postconfluent cells enhanced ER-binding capacity in a dose-dependent manner and estrogen responsiveness of an exogenous reporter gene and of the endogenous alkaline phosphatase, representing an endogenous estrogen-stimulated gene. These data indicate that there is an interaction between the PKC and ER signaling systems in bone cells and that this interaction may be influenced by the proliferative and/or differentiative state of the cells, resulting in modulation of hormone responsiveness. PMID- 8247016 TI - Regulation of endogenous thyrotropin-releasing hormone receptor messenger RNA in GH4C1 cells: roles of protein and RNA synthesis. AB - In previous studies on the regulation of the endogenous TRH receptor (TRHR) in GH4C1 cells, we have shown that the synthetic glucocorticoid dexamethasone (Dex) increased TRHR abundance on the plasma membrane of these cells as well as increasing the concentration of TRHR mRNA. In the present investigation, we determined whether the action of Dex on TRHR mRNA was direct or whether it involved the synthesis of intermediary protein(s). We found that the protein synthesis inhibitors cycloheximide or anisomycin alone enhanced TRHR mRNA accumulation and, in the presence of Dex, caused a 10- to 20-fold superinduction of TRHR mRNA. The superinduction required inhibition of protein synthesis. Results of nuclear run-on assays showed that superinduction was accompanied by a large increase in transcription of TRHR mRNA. Experiments designed to examine the stability of TRHR mRNA transcripts revealed that inhibitors of RNA synthesis (actinomycin D and dichlorobenzimidazole riboside) caused an increase in endogenous TRHR mRNA concentrations by themselves, complicating their use to estimate TRHR mRNA half-life. We conclude that Dex stimulates transcription of endogenous TRHR mRNA by a mechanism that does not require enhanced synthesis of new proteins. In fact, ongoing protein synthesis reduces steady state TRHR mRNA concentrations consistent with a role for a labile protein(s) in suppressing TRHR gene transcription; an additional action of a labile protein to enhance TRHR mRNA degradation is also possible. PMID- 8247017 TI - Molecular cloning of mouse pancreatic islet R-cadherin: differential expression in endocrine and exocrine tissue. AB - A search for novel pancreatic islet cadherins was undertaken using the polymerase chain reaction with mouse beta TC3 cell line cDNA and degenerate primers based on conserved C-terminal sequences in neural (N), epithelial, and placental cadherin (CAD). A hitherto uncharacterized rodent sequence was detected which was then cloned from a mouse insulinoma cDNA library and shown to be the mouse equivalent of chicken retina CAD (R-CAD). The similarity of the mouse and chicken sequences was remarkable (eight nonconservative changes in the 747 amino acids of the mature protein sequence; 95% overall identity), indicating strong conservation of function. Mouse R-CAD was also closely homologous to N-CAD (72% identity), including those regions of N-CAD implicated in the cadherin-cadherin interaction and Ca2+ binding. In vitro translation of the cDNA indicated that mouse R-CAD enters the secretory pathway and undergoes posttranslational glycosylation and proteolytic cleavage. R-CAD mRNA was distributed widely in mouse tissues with high levels present in brain, skeletal muscle, and thymus. In the pancreas, R-CAD and N-CAD showed endocrine cell specificity and a differential expression in beta and non-beta-cells. Messenger RNA expression was evident during early pancreatic development at a time when the first pluripotent hormone-producing cells differentiate to attain their adult phenotype and become organized in islet-like clusters. The presence of R-CAD and N-CAD in islets is consistent with the neurone-like properties of this tissue. Differences in CAD expression might explain the segregation of exocrine and endocrine cells during development of the pancreas and the characteristic morphological distribution of the different endocrine cells within the islet. PMID- 8247018 TI - Basic fibroblast growth factor stimulates DNA synthesis in cells overexpressing the insulin-like growth factor-I receptor. AB - Insulin-like growth factor-I (IGF-I), by itself, cannot sustain the growth of BALB/c 3T3 cells, but requires the cooperation of other growth factors, such as platelet-derived growth factor or epidermal growth factor. In 3T3 cells constitutively overexpressing the human IGF-I receptor, called p6 cells, IGF-I by itself is fully mitogenic. We show here that p6 cells are also stimulated to enter DNA synthesis by the sole addition of basic fibroblast growth factor (bFGF), which, by itself, is incapable of stimulating parental 3T3 cells. Although bFGF does not bind directly to the IGF-I receptor, it induces its autophosphorylation. Stimulation of p6 cells by bFGF is not inhibited by an antibody to the IGF-I receptor that inhibits IGF-I-mediated DNA synthesis, and IGF-I is not detectable in the medium of bFGF-treated p6 cells. Stimulation cannot be explained by an increased number of FGF receptors, because p6 cells actually have slightly fewer FGF receptors than parental BALB/c 3T3 cells. Basic FGF also stimulates DNA synthesis in 3T3 cells overexpressing a mutant IGF-I receptor that does not autophosphorylate in response to IGF-I and has lost its mitogenic potential. Although we were unable to demonstrate directly that bFGF causes transphosphorylation of the IGF-I receptor, we conclude that in cells overexpressing the IGF-I receptor, bFGF can stimulate DNA synthesis either by an unknown mechanism or through transphosphorylation of the IGF-I receptor. PMID- 8247019 TI - Mechanism of glucocorticoid induction of the rat plasminogen activator inhibitor 1 gene in HTC rat hepatoma cells: identification of cis-acting regulatory elements. AB - Type 1 plasminogen activator inhibitor (PAI-1) is the major physiological inhibitor of plasminogen activation, inhibiting both tissue- and urokinase-type plasminogen activators. In HTC rat hepatoma cells, glucocorticoids increase PAI-1 activity, antigen and mRNA accumulation 3- to 5-fold; this increase is due solely to an increase in the rate of PAI-1 gene transcription. We have identified the cis-acting sequences in the 5'-flanking sequence of the HTC PAI-1 gene that mediate this induction. Analysis of a series of hybrid genes containing various portions of the PAI-1 5'-flanking region fused to the chloramphenicol acetyltransferase reporter gene transfected into HTC cells localized the region involved in the transcriptional regulation by glucocorticoids to between -1237 and -764. Electrophoretic mobility shift assays and DNase-I protection assays showed that a glucocorticoid response element (GRE) 15-mer located at -1212 bound the glucocorticoid receptor DNA-binding domain protein in a concentration dependent manner. Mutations created within this GRE eliminated its ability both to confer a glucocorticoid response and to bind the glucocorticoid receptor. When placed upstream of a heterologous promoter in either orientation, this GRE conferred glucocorticoid inducibility. We, therefore, conclude that the sole cis acting sequence required for the glucocorticoid response of the PAI-1 gene in rat HTC hepatoma cells is the GRE at -1212. PMID- 8247020 TI - Roles of extracellular and cytoplasmic domains of the prolactin receptor in signal transduction to milk protein genes. AB - We have previously shown that the long form of the PRL receptor is able to activate milk protein gene transcription. In the present study, we have determined the respective contribution of the extracellular and the intracellular domains of this receptor to transcriptional activation of a milk protein gene by PRL. The membrane-anchored intracellular domain (pTMI) expressed alone was devoid of PRL binding activity, as expected, and did not constitutively stimulate expression of the target gene. The extracellular domain (pE), expressed alone as a soluble receptor form, binds PRL with 10-fold higher affinity than the full length membrane receptor. This form was also unable to stimulate the expression of the reporter gene. However, expression of both mutants (pE + pTMI) in the same cell partially restored the ability of PRL to activate the beta-lactoglobulin promoter. Replacement of cysteine 184 by a serine in the extracellular domain of the receptor impairs this restoration of the biological response. However, introduction of the same mutation in the full-length receptor did not affect its functional activity. These results indicate that the membrane-anchored cytoplasmic domain of the PRL receptor has no constitutive activity, and that coexpression of individual extracellular and intracellular domains leads to restoration of receptor function. We propose that restoration may be the result of reconstitution of the holoreceptor through disulfide bonding, or it may be the result of interaction of the extracellular region with an external transducing molecule. PMID- 8247021 TI - Functional analysis of the amino acids in the DNA recognition alpha-helix of the human thyroid hormone receptor. AB - The roles in DNA binding and transcriptional activation of individual amino acids in the putative recognition alpha-helix of the first zinc finger of the beta isoform of the human thyroid hormone receptor (hT3R beta) have been probed by site-directed mutagenesis. Alanine substitutions of highly conserved residues involved in the folding of this zinc finger abolished the binding of hT3R beta to various thyroid response elements. A similar effect was observed for alanine substitutions of those conserved residues in hT3R beta that were expected to make specific contacts to DNA bases common to all hormone response elements. The three P-box amino acids have previously been shown to be essential for discrimination of the base pairs that differ between the DNA binding sites for related steroid/thyroid hormone receptors. In hT3R beta, the P-box residues are E, G, and G; the results of this study show that alanine substitution of the glutamic acid dramatically reduces DNA binding activity by hT3R beta, while the substitution of either glycine has little or no effect on DNA binding. The effects of alanine substitutions on hT3R beta transcriptional activation properties were consistent with the effect of these substitutions on DNA binding properties, with the exception of the second P-box amino acid. T3R beta substituted with alanine at this position is substantially more defective in transcriptional activation than it is in specific DNA binding. These results indicate that there are two separate mechanisms of response element discrimination by P-box amino acids of steroid/thyroid hormone receptors, one which operates at the level of DNA recognition and a second which operates at the level of transcriptional activation. PMID- 8247022 TI - Activation of CYP11A and CYP11B gene promoters by the steroidogenic cell-specific transcription factor, Ad4BP. AB - We have examined the transcriptional activity of four cis-elements, Ad1(CRE), Ad2, Ad3, and Ad4, that are present in the promoter of the bovine CYP11B (11 beta hydroxylase P-450) gene using beta-globin reporter gene constructs and transient transfection into steroidogenic and nonsteroidogenic cell types. Only Ad1(CRE), a CRE homolog, showed forskolin-dependent transcriptional activity in adrenal tumor Y-1 cells, whereas the other elements were not able to stimulate transcription by themselves. As Ad3 and Ad4 had previously been identified as the cis-elements required for full cAMP-dependent transcription of this gene, we examined the effect of combinations of different cis-elements on the transcription of the reporter gene. In Y-1 cells, Ad1(CRE) and four tandem copies of any one of the other cis-elements substantially activated transcription in response to forskolin treatment. The template carrying Ad1(CRE) and Ad4 was also active in testicular Leydig cells, I-10, whereas it was inactive in nonsteroidogenic PC-12 cells. Transcriptional activation by the 4xAd4/Ad1(CRE) combination presumably depended on the presence of Ad4-binding protein (Ad4BP), which is absent in PC-12 cells, as shown by immunoblot analysis. This was confirmed by cotransfecting an expression vector for Ad4BP into PC-12 cells, which caused forskolin-dependent transcription to increase in proportion to the amount of expression vector. In Y 1 cells, transcriptional activation by forskolin was mimicked by cotransfection of an expression vector for the catalytic subunit of protein kinase-A.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247023 TI - Identification of 3',5'-cyclic adenosine monophosphate-inducible nuclear factors binding to the human urokinase promoter in mouse Sertoli cells. AB - Transcription factors which recognize both the SV40 promoter and the proximal promoter region of the human urokinase-type plasminogen activator (h-uPA) gene are present in nuclear extracts from primary cultures of mouse Sertoli cells; prolonged (more than 12 h) (Bu)2cAMP stimulation of Sertoli cells induces the formation of different specific DNA-protein complexes. A discrete region in the h uPA promoter, between -54 and -42, is essential for the formation of the cAMP induced DNA-protein complexes. Mutation of the sequence between -54 and -42 abolishes the response to cAMP of the proximal h-uPA promoter in Sertoli cells. A protein, recognizing a sequence centered around the GC-box present between -48 and -43, is detected by Southwestern analysis, and it is clearly induced by (Bu)2cAMP stimulation. Interaction between this protein and a second factor, recognizing a purine-rich sequence between -53 and -46, partially overlapping the GC-box, is needed for the formation of the cAMP-induced DNA-protein complexes. A preformed complex between the cAMP-induced GC-box-binding factor and the second factor can be detected using nondenaturing conditions during Southwestern analysis. PMID- 8247024 TI - Demonstration of nuclear translocation of the mineralocorticoid receptor (MR) using an anti-MR antibody and confocal laser scanning microscopy. AB - Using a synthetic peptide that corresponds to a unique region of the N-terminal domain of the human mineralocorticoid receptor (MR), amino acids 87-96, we have generated a polyclonal antibody, human (h) MRsN. This sequence shares no homology with the corresponding sequences of the glucocorticoid receptor or other steroid/thyroid hormone receptor superfamily members. Antibody hMRsN cross-reacts with MR from human, rat, and mouse cells and recognizes denatured MR from either crude preparations or partially purified rat kidney cytosol, rat colon, or recombinant hMR overexpressed in baculovirus-infected Sf9 cells. Immunoprecipitation of the native MR from either partially purified or crude preparations of rat kidney cytosol with hMRsN, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and silver stain, demonstrated a major protein band with a mol wt of 116 kilodaltons. In addition, using confocal laser scanning microscopy and digital image analysis, the immunocytochemical localization of the recombinant hMR over-expressed in Sf9 cells 24 h post-transfection was determined. In the absence of ligand, the MR was detected solely in the cytoplasm, after a 30-min exposure to 100 nM aldosterone the MR was perinuclear, and after 60 min, the MR was predominantly nuclear. To ascertain that this phenomenon was not unique to insect cells, aldosterone induced MR nuclear translocation in mouse macrophage cells was also demonstrated immunocytochemically, clearly indicating a role for nuclear translocation in MR function. PMID- 8247025 TI - Molecular cloning and expression of an abundant rabbit ovarian protein with 20 alpha-hydroxysteroid dehydrogenase activity. PMID- 8247026 TI - Immunodominance of a recombinant T-cell epitope depends on its molecular environment. AB - In the present study, we have investigated the influence of the molecular environment of a T-cell epitope on its immunogenicity. We genetically inserted into different sites of two bacterial recipient proteins, LamB or MalE, an immunodominant T-cell epitope: the 120-132 T-cell epitope from the PreS2 region of HBV. The T-cell epitope was introduced, either alone (PreS:T) or with an adjacent B-cell epitope (PreS:TB). After purification, the hybrid proteins were injected into mice and we studied the immunogenicity of recombinant T-cell epitopes by analyzing the in vitro proliferative responses of LN cells from these mice to the inserted peptides. The immunization of mice with recombinant MalE protein containing the PreS:T or PreS:TB peptides at two different sites induced strong peptide-specific proliferative responses, indicating that the insertion sites did not affect the immunodominance of the inserted T-cell epitope. A strong T-cell proliferative response was also obtained after immunization of mice with hybrid LamB protein containing the PreS:TB epitope at position 153. In contrast, the recombinant proteins which contained only the PreS:T epitope at positions 153 or 374 failed to stimulate T-cell responses. Therefore, this study demonstrates that the immunogenicity of recombinant T-cell epitopes may be strongly affected both by the insertion site and by inserted adjacent residues. PMID- 8247027 TI - Specific V gene usage in anti-phosphorylcholine IgE antibodies. AB - Three hybridomas from phosphorylcholine(PC)-KLH immunized BALB/c mice producing IgE antibodies against the PC hapten were investigated for their fine specificity to the hapten and usage of V gene segments in H- and L-chains. All three IgE antibodies recognize the entire azophenyl-PC hapten. They are T15 Id negative and do not bind to the natural PC determinant expressed by the Streptococcus carbohydrate R36A. T15 Id positive IgE antibodies could neither be elicited by immunization in detectable amounts nor generated by the cell fusion technique. By using the Southern blot technique and nucleotide sequence analysis of PCR amplified VHDJH and VLJL rearrangements, we have demonstrated that the three IgE anti-PC hybridomas use the VH1-DSP2-JH2, the VHOX1-DSP2-JH3 or the VH36-60-D-JH2 gene segment combinations for the H chain together with the V kappa 1C-J kappa 1, V kappa 1C-J kappa 2 or V lambda 1-J lambda 1 genes for the L chains. Except for the VH36-60, the same gene segments were found in different combinations in anti PC antibodies of other Ig classes than IgE. However, high rates of somatic mutations are expressed in both VH1 of the H chain and in V kappa 1C of the L chain. The VH36-60 is expressed in antibodies with the major Id of the azophenyl arsonate (Ars) response and VHOX1 generally contributes to the phenyl-oxazolone specificity. This suggests that these V genes are involved in the recognition of the azophenyl moiety of the coupled PC hapten. Thus PC-KLH specific IgE antibodies utilize mutated VH1 and/or VH/VL gene segment combinations which are involved in binding of the azophenyl spacer. These IgE are therefore specific for azophenyl-phosphorylcholine, unlike antibodies normally expressed against the Streptococcus PC determinant in mice. The genetic diversity and the high mutation rates indicate that the specific B cells develop later in the immune response. Thus, they represent newly generated specificities of so-called group II anti-PC antibodies and are not isotype-switch descendants from already existing T15 Id positive IgM antibodies. PMID- 8247028 TI - Enhancement of HIV-1 gp41 binding to Raji cells by PWM, LPS, interferon-gamma and interleukin-6. AB - Based on our finding that HIV-1 gp41 independently of CD4 can bind to several proteins (gp41 binding protein: GBP) on the human T-cell line H9, B-cell line Raji and monocyte-cell line U937, we examined the effect of mitogens and cytokines on binding of gp41 to H9, Raji and U937 cells. Flow cytometry (FACS) analysis demonstrated that PWM and LPS, IFN-gamma and IL-6, but not Con A, IFN alpha, -beta, -omega and IL-2, could increase gp41 binding to Raji cells. In controls, none of the regulators (IFN-alpha, -beta, -gamma, -omega, IL-2, IL-6, Con A, PWM, LPS) could modify the binding potential of H9 and U937 cells. Our data suggest that the expression of HIV-1 binding proteins is subject to regulation by PWM, LPS, IFN-gamma and IL-6 in the case of B-cells, while on T cells and macrophages, the binding proteins may be constitutively expressed. PMID- 8247029 TI - Human complement factor B: cDNA cloning, nucleotide sequencing, phenotypic conversion by site-directed mutagenesis and expression. AB - A full-length cDNA clone, BHL4-1, encoding factor B was isolated from a human liver cDNA library and sequenced in its entirety. It consists of 2388 bp which include a 5'-untranslated region of 40 bp, a single open reading frame, 2292 bp in length, and a 3'-untranslated region of 56 bp followed by a poly-A tail. The deduced amino acid sequence comprises 25 residues of a putative leader peptide and 739 residues of the mature polypeptide chain of the F allele of factor B. We constructed an S allele-like Q7R mutant of BHL4-1 by site-directed mutagenesis. Both the wild-type and mutant factor B cDNA were expressed transiently in a eukaryotic system. The specific hemolytic activities of the two recombinant factor B alleles and of native B were not significantly different from each other. PMID- 8247030 TI - Membrane immunoglobulin is characterized by distinct structural subpopulations. AB - Membrane immunoglobulins are integral proteins on B cell surfaces that bind foreign antigens and are critically involved in the regulation of the immune response. Based upon the model of serum IgG, it has been assumed that membrane immunoglobulins are essentially four chain disulfide-linked structures of the form H2L2, where H represents an immunoglobulin heavy chain, and L a light chain. We show here that membrane immunoglobulins of the mu and delta isotypes are present on spleen cell surfaces in a much more diverse group of disulfide linked structures. In some cases mIg is linked into structures as large as H5L5, while in other instances mu or delta chains appear to be linked by disulfide interactions to non-immunoglobulin molecules. These various structural complexes may represent distinct functional entities, as the association of mIg with the cytoskeleton after mIg cross-linking appears to depend upon its structural subgroup. PMID- 8247031 TI - Molecular characterization of human antibodies to bacterial antigens: utilization of the less frequently expressed VH2 and VH6 heavy chain variable region gene families. AB - Structural analysis of the human immunoglobulin repertoire holds promise for determining the basis of variable region gene usage in response to a variety of auto and exogenous antigens. Here we report the nucleotide sequences of the heavy and light chain variable regions expressed by three human monoclonal antibodies specific for two clinically relevant bacterial pathogens, Bordetella pertussis and Haemophilus influenzae type b. The cell lines were derived by in vitro stimulation of lymphocytes from spleen or tonsillar tissue, respectively, and bind to different antigens from the two organisms. The single B. pertussis antibody is of the IgM lambda isotype and utilizes the single VH6 gene segment in combination with a V lambda 2 gene and demonstrates limited somatic mutation, yet is highly indicative of an antigen-driven immune response. One H. influenzae antibody is of the IgG2 lambda isotype and expresses a VH3 gene segment with a V lambda 1 gene, while the second cell line produces an IgG3 lambda antibody expressing a combination of VH2/V lambda 3. Both molecules show evidence of somatic mutation. The D gene segments of the heavy chains vary in length and display limited sequence homology with known germline D segments. As demonstrated previously, JH4 predominates (two JH4 and one JH3) and all three utilize the J lambda 3 gene segment. In addition, we have isolated and sequenced a number of germline VH2 gene segments in an attempt to better understand the nature of the VH2 germline repertoire. In addition to contributing to the understanding of the human antibody repertoire, such clinically relevant molecules may prove to be a source of passive immunotherapy for those at risk to developing disease. PMID- 8247032 TI - Rearrangement by chromosomal inversion in the T cell receptor gamma locus in a murine alpha beta T cell clone. AB - Analyzing the T-cell receptor (TCR) repertoire of T lymphocytes specific for minor histocompatibility antigens, we isolated a TCR-alpha beta bearing T cell clone, TGVH9, which also expresses three different TCR-gamma mRNA. The sequences of these messengers indicate that one of them results from the rearrangement of V gamma 1 and J gamma 2 genes. Since these two segments are in inverted genomic transcriptional orientation, the joining of V gamma 1 to J gamma 2 occurred by chromosomal inversion. This mechanism, which was previously reported in TCR-beta and TCR-delta loci, can thus also occur to enlarge the diversity of the TCR-gamma repertoire. PMID- 8247033 TI - Moderate alcohol intake, increased levels of high-density lipoprotein and its subfractions, and decreased risk of myocardial infarction. AB - BACKGROUND: Previous studies have suggested that moderate alcohol intake exerts a protective effect against coronary heart disease. Alterations in plasma lipoprotein levels represent one plausible mechanism of this apparent protective effect. METHODS: We therefore examined the interrelation among alcohol consumption, plasma lipoprotein levels, and the risk of myocardial infarction in 340 patients who had had myocardial infarctions and an equal number of age- and sex-matched controls. The case patients were men or women less than 76 years of age with no history of coronary disease who were discharged from one of six hospitals in the Boston area with a diagnosis of a confirmed myocardial infarction. Alcohol consumption was estimated by means of a food-frequency questionnaire. RESULTS: We observed a significant inverse association between alcohol consumption and the risk of myocardial infarction (P for trend, < 0.001 after control for known coronary risk factors). In multivariate analyses, the relative risk for the highest intake category (subjects who consumed three or more drinks per day) as compared with the lowest (those who had less than one drink a month) was 0.45 (95 percent confidence interval, 0.26 to 0.80). The levels of total high-density lipoprotein cholesterol (HDL) and its HDL2 and HDL3 subfractions were strongly associated with alcohol consumption (P for trend, < 0.001 for each). The addition of HDL or either of its subfractions to the multivariate model substantially reduced the inverse association between alcohol intake and myocardial infarction, whereas the addition of the other plasma lipid measurements did not materially alter the relation. CONCLUSIONS: These data confirm the inverse association of moderate alcohol intake with the risk of myocardial infarction and support the view that the effect is mediated, in large part, by increases in both HDL2 and HDL3. PMID- 8247034 TI - Transmission from one child to another of human immunodeficiency virus type 1 with a zidovudine-resistance mutation. AB - BACKGROUND AND METHODS. We describe a child who apparently acquired human immunodeficiency virus type 1 (HIV-1) infection in the home setting. The suspected source of infection was a child with the acquired immunodeficiency syndrome who had received zidovudine and whose virus contained a mutation associated with in vitro zidovudine resistance. The children were born to different HIV-1-infected mothers, but they lived in the same home between the ages of two and five years. Child 1 was infected perinatally; Child 2 was not and was repeatedly found to be seronegative. Child 2 was examined because of acute lymphadenopathy and had seroconverted to HIV-1 positivity. HIV-1 proviral DNA was amplified from peripheral-blood mononuclear cells and subjected to sequence analysis. Sequences from Child 2 were compared with those from Child 2's mother, Child 1, and local HIV-1-infected control children. PMID- 8247035 TI - Liver transplantation in European patients with the hepatitis B surface antigen. AB - BACKGROUND: The role of liver transplantation in patients positive for the hepatitis B surface antigen (HBsAg) is controversial because of the high rate of recurrent hepatitis B virus (HBV) infection. It has not been determined whether this risk is greater for certain patients and whether the administration of anti hepatitis B surface antigen (anti-HBs) immune globulin is beneficial. METHODS: We conducted a retrospective study at 17 European centers of 372 consecutive HBsAg positive patients who underwent liver transplantation between 1977 and 1990. Recurrence of HBV infection was defined as the reappearance of HBsAg in serum. RESULTS: For all 334 patients with follow-up data, the mean (+/- SE) three-year actuarial risk of recurrence of HBV was 50 +/- 3 percent. The risk was 67 +/- 4 percent among 163 patients with HBV-related cirrhosis, 32 +/- 5 percent among 110 patients with cirrhosis related to hepatitis delta virus, 40 +/- 16 percent among 14 patients with fulminant hepatitis delta infection, and 17 +/- 7 percent among 39 patients with fulminant HBV infection (P < 0.001). Among the patients with HBV related cirrhosis, the risk of HBV recurrence was greatest (83 +/- 6 percent) in those who were seropositive for HBV DNA at the transplantation and lowest (58 +/- 7 percent) in those with neither HBV DNA nor hepatitis B e antigen detectable in serum. With respect to the use of passive prophylaxis with anti-HBs immune globulin, the risk of HBV recurrence was 75 +/- 6 percent among the 67 patients given no immunoprophylaxis, 74 +/- 5 percent among the 83 treated for two months, and 36 +/- 4 percent among the 209 treated for six months or longer (P < 0.001). In a multivariate analysis the predictors of a lower risk of HBV recurrence were the long-term administration of the immune globulin, hepatitis delta virus superinfection, and acute liver disease. For the entire study cohort, survival was 75 percent at one year and 63 percent at three years, but for those in whom HBV infection recurred, survival was 68 percent at one year and 44 percent at three years. CONCLUSIONS: In this retrospective study of HBsAg-positive patients, liver transplantation had better results in those who had fulminant hepatitis or delta virus superinfection. An absence of viral replication at the time of transplantation and long-term immunoprophylaxis were associated with a reduced risk of recurrent HBV infection and reduced mortality. PMID- 8247036 TI - Initial chemotherapeutic doses and survival in patients with limited small-cell lung cancer. AB - BACKGROUND: Moderate increases in the initial doses of certain chemotherapeutic drugs, such as cisplatin and cyclophosphamide, may prolong overall survival in patients with limited small-cell lung cancer. METHODS: We conducted a prospective study of 105 patients with limited small-cell lung cancer. The patients were randomly assigned to receive higher or lower initial doses of cisplatin (100 or 80 mg per square meter of body-surface area) and cyclophosphamide (300 or 225 mg per square meter daily for four days); all patients received the same doses of doxorubicin and etoposide. The first course of chemotherapy was followed by five additional courses and by three courses of radiotherapy. All patients received the lower doses of cisplatin and cyclophosphamide and the same doses of doxorubicin and etoposide from the second through the sixth cycle of chemotherapy. RESULTS: The median follow-up was 33 months. The two-year survival rate for the 55 patients who received the higher doses of chemotherapy was 43 percent, as compared with 26 percent for the 50 patients who received the lower doses (P = 0.02). The rates of complete response at six months were 67 percent in the higher-dose group and 54 percent in the lower-dose group (P = 0.16). Disease free survival at two years was 28 percent in the higher-dose group, as compared with 8 percent in the lower-dose group (P = 0.02). Side effects from treatment were not increased in the higher-dose group. CONCLUSIONS: Higher initial doses of cyclophosphamide and cisplatin improve disease-free and overall survival in patients with limited small-cell lung cancer. PMID- 8247037 TI - Lack of immune responsiveness to bovine serum albumin in insulin-dependent diabetes. AB - BACKGROUND: Epidemiologic studies have implicated the ingestion of cow's milk in the pathogenesis of insulin-dependent diabetes mellitus (IDDM). Moreover, in a recent study, 100 percent of patients with new-onset IDDM had antibodies against bovine serum albumin (BSA), with a majority directed against a 17-amino-acid BSA peptide (ABBOS). Cellular immune mechanisms are thought to be the principal mediators of pancreatic beta-cell destruction in IDDM. METHODS: We measured the responses of peripheral-blood mononuclear cells to BSA and ABBOS or serum IgG anti-BSA antibodies (by particle-concentration fluorescence immunoassay) in 71 patients with IDDM, 55 subjects at various degrees of risk for IDDM, 36 patients with other autoimmune disorders (chronic autoimmune thyroiditis, rheumatoid arthritis, and systemic lupus erythematosus), and 48 normal subjects. RESULTS: The responses of peripheral-blood mononuclear cells to BSA or ABBOS were positive in 2 of 24 patients with new-onset IDDM, 1 of 25 first-degree relatives of patients with IDDM who were negative for islet-cell antibodies, 2 of 30 first degree relatives of patients with IDDM who were positive for islet-cell antibodies, 1 of 28 patients with established IDDM, and 1 of 29 normal subjects. Similarly, anti-BSA antibodies were not detected significantly more often in patients with new-onset IDDM (3 of 31, 10 percent) than in normal subjects (1 of 37, 3 percent; P = 0.32). However, many patients with autoimmune disease and subjects at increased risk for IDDM had anti-BSA antibodies (frequency, 10 to 31 percent). CONCLUSIONS: Anti-BSA antibodies may reflect a general defect in the process of immunologic tolerance associated with a predisposition to autoimmunity rather than immunity specific to beta cells. The absence of cellular immunity to BSA and ABBOS in IDDM does not support a role for this antigen in the pathogenesis of the disorder. PMID- 8247038 TI - Brief report: herpetic geometric glossitis. PMID- 8247039 TI - Images in clinical medicine. Clubbing and hypertrophic osteoarthropathy. PMID- 8247040 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 50-1993. A 46-year-old man with postobstructive pneumonia and a pulmonary mass. PMID- 8247041 TI - Is alcohol good for your health? PMID- 8247042 TI - HIV prevention--bringing the message home. PMID- 8247043 TI - Hepatitis B and liver transplantation. Problems and promises. PMID- 8247044 TI - Cardiology--division or department? PMID- 8247046 TI - Coronary atherectomy versus angioplasty. PMID- 8247045 TI - Coronary atherectomy versus angioplasty. PMID- 8247047 TI - Number of pregnancies and risk of cardiovascular disease. PMID- 8247048 TI - Number of pregnancies and risk of cardiovascular disease. PMID- 8247049 TI - Zidovudine in asymptomatic HIV infection. PMID- 8247050 TI - Community-acquired methicillin-resistant Staphylococcus aureus infection. PMID- 8247051 TI - Risk factors for hepatocellular carcinoma in patients with chronic liver diseases. PMID- 8247052 TI - Autoimmune protein S deficiency. PMID- 8247053 TI - Spontaneous expectoration of a prosthetic graft nine years after a Blalock Taussig shunt procedure. PMID- 8247054 TI - Anal-sphincter disruption during vaginal delivery. AB - BACKGROUND: Lacerations of the anal sphincter or injury to sphincter innervation during childbirth are major causes of fecal incontinence, but the incidence and importance of occult sphincter damage during routine vaginal delivery are unknown. We sought to determine the incidence of damage to the anal sphincter and the relation of injury to symptoms, anorectal physiologic function, and the mode of delivery. METHODS: We studied 202 consecutive women six weeks before delivery, 150 of them six weeks after delivery, and 32 with abnormal findings six months after delivery. Symptoms of anal incontinence and fecal urgency were assessed, and anal endosonography, manometry, perineometry, and measurement of the terminal motor latency of the pudendal nerves were performed. RESULTS: Ten of the 79 primiparous women (13 percent) and 11 of the 48 multiparous women (23 percent) who delivered vaginally had anal incontinence or fecal urgency when studied six weeks after delivery. Twenty-eight of the 79 primiparous women (35 percent) had a sphincter defect on endosonography at six weeks; the defect persisted in all 22 women studied at six months. Of the 48 multiparous women, 19 (40 percent) had a sphincter defect before delivery and 21 (44 percent) afterward. None of the 23 women who underwent cesarean section had a new sphincter defect after delivery. Eight of the 10 women who underwent forceps delivery had sphincter defects, but none of the 5 women who underwent vacuum extractions had such defects. Internal sphincter defects were associated with a significantly lower mean (+/- SD) resting anal pressure (61 +/- 11 vs. 48 +/- 10 mm Hg, P < 0.001) six weeks post partum, and external-sphincter defects were associated with a significantly lower squeeze pressure (increase above resting pressure, 70 +/- 38 vs. 44 +/- 13 mm Hg; P < 0.001). There was a strong association (P < 0.001) between sphincter defects and the development of bowel symptoms. CONCLUSIONS: Occult sphincter defects are common after vaginal delivery, especially forceps delivery, and are often associated with disturbance of bowel function. PMID- 8247055 TI - The natural history of borderline isolated systolic hypertension. AB - BACKGROUND: Patients with isolated systolic hypertension are at increased risk for cardiovascular disorders. We attempted to determine whether those with borderline isolated systolic hypertension (defined as a systolic blood pressure of 140 to 159 mm Hg and a diastolic blood pressure below 90 mm Hg) have a greater risk of progression to definite (more severe) hypertension and of major morbid or fatal events than people with normal blood pressure (< 140/90 mm Hg). METHODS: A total of 2767 of the original participants in the Framingham Heart Study were monitored with biennial examinations for up to 34 years for the development of definite hypertension (defined as a systolic blood pressure of > or = 160 mm Hg, a diastolic blood pressure of > or = 90 mm Hg, or the initiation of antihypertensive therapy) and for major cardiovascular events. RESULTS: Borderline isolated systolic hypertension was the most common type of untreated hypertension among adults over the age of 60. After 20 years of follow-up, 80 percent of those with borderline isolated systolic hypertension had progression to definite hypertension, as compared with 45 percent of the normotensive participants (P < 0.001). After adjustment for age, sex, and risk factors for cardiovascular disease, participants with borderline isolated systolic hypertension had an excess long-term risk of cardiovascular disease (hazard ratio, 1.47; 95 percent confidence interval, 1.24 to 1.74) and death from cardiovascular disease (hazard ratio, 1.57; 95 percent confidence interval, 1.24 to 2.00), as compared with normotensive participants. In an analysis of pooled data from biennial examinations to study short-term sequelae, subjects with borderline isolated systolic hypertension had an increased risk of progression to definite hypertension (odds ratio, 3.84; 95 percent confidence interval, 3.35 to 4.41) and of cardiovascular disease (odds ratio, 1.39; 95 percent confidence interval, 1.06 to 1.82). CONCLUSIONS: In both the short term and the long term, subjects with borderline isolated systolic hypertension are at increased risk of progression to definite hypertension and the development of cardiovascular disease. PMID- 8247056 TI - Renal tubular dysfunction in chronic alcohol abuse--effects of abstinence. AB - BACKGROUND: Alcohol abuse may be accompanied by a variety of disorders of electrolyte and acid-base metabolism. The role of the kidney in the pathogenesis of these disturbances is obscure. We sought to evaluate the alcohol-induced abnormalities of renal function and improvement during abstinence and to assess the relation between renal dysfunction and electrolyte and acid-base disorders. METHODS: We measured biochemical constituents of blood and renal function before and after four weeks of abstinence in 61 patients with chronic alcoholism who had little or no liver disease. RESULTS: On admission, 18 patients (30 percent) had hypophosphatemia and hypomagnesemia, 13 patients (21 percent) had hypocalcemia, and 8 patients (13 percent) had hypokalemia. Twenty-two patients (36 percent) had a variety of simple and mixed acid-base disorders. Twenty of these patients had metabolic acidosis, and among them, 80 percent had alcoholic acidosis. A wide range of defects in renal tubular function, with normal glomerular filtration rate, were detected in these patients. The defects included decreases in the threshold and maximal reabsorptive ability for glucose (38 percent of patients) and in the renal threshold for phosphate excretion (36 percent); increases in the fractional excretion of beta 2-microglobulin (38 percent), uric acid (12 percent), calcium (23 percent), and magnesium (21 percent); and aminoaciduria (38 percent). Seventeen patients (28 percent) had a defect in tubular acidification, and five an impairment in urinary concentrating ability. Urinary excretion of N acetyl-beta-D-glucosaminidase and alanine aminopeptidase were increased in 41 and 34 percent of patients, respectively. The abnormalities of blood chemistry and renal tubular function disappeared after four weeks of abstinence. CONCLUSIONS: Transient defects in renal tubular function are common in patients with chronic alcoholism and may contribute to their abnormalities of serum electrolyte and blood acid-base profiles. PMID- 8247057 TI - Images in clinical medicine. Coccidioides immitis. PMID- 8247058 TI - Sexual harassment of female doctors by patients. AB - BACKGROUND: Sexual harassment within the doctor-patient relationship is typically discussed in terms of male doctors harassing female patients. We investigated the sexual harassment of female doctors by patients. METHODS: Surveys were mailed to a random sample of 599 of the 1064 licensed female family physicians in Ontario, Canada. Respondents were asked about their experiences of sexual harassment by either male or female patients and about the nature and frequency of harassing behavior. Suggestions for prevention were requested. RESULTS: Seventy percent (422) of the questionnaires were completed and returned. More than 75 percent of the respondents reported some sexual harassment by patient at some time during their careers. Physicians had been harassed most often in their own offices and by their own patients. However, in settings such as emergency rooms and clinics, unknown patients presented a proportionately higher risk. The physicians' perceptions of the seriousness of the problem varied with the frequency and severity of the incidents. CONCLUSIONS: Sexual harassment of female doctors appears to occur frequently, and it is therefore an important topic to address in medical school and professional development. PMID- 8247059 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 51-1993. A 36-year-old man with AIDS, increase in chronic diarrhea, and intermittent fever and chills. PMID- 8247060 TI - Childbirth, continence, and the pelvic floor. PMID- 8247061 TI - The Comprehensive Childhood Immunization Act of 1993. Toward a more rational approach. PMID- 8247062 TI - Changes in the mix of AIDS-defining conditions. PMID- 8247063 TI - Maternal diet and primitive neuroectodermal brain tumors in children. PMID- 8247064 TI - Brain tumors in infants. PMID- 8247065 TI - Brain tumors in infants. PMID- 8247066 TI - Aortic-valve function during cardiopulmonary resuscitation. PMID- 8247067 TI - Endothelin-1 in pulmonary hypertension. PMID- 8247068 TI - More on vaccine-associated paralytic poliomyelitis. PMID- 8247069 TI - Toward a social policy for health. PMID- 8247070 TI - Trans-fatty-acid content of common foods. PMID- 8247071 TI - A student's view of a medical teaching exercise. PMID- 8247072 TI - Prevention of colorectal cancer by colonoscopic polypectomy. The National Polyp Study Workgroup. AB - BACKGROUND: The current practice of removing adenomatous polyps of the colon and rectum is based on the belief that this will prevent colorectal cancer. To address the hypothesis that colonoscopic polypectomy reduces the incidence of colorectal cancer, we analyzed the results of the National Polyp Study with reference to other published results. METHODS: The study cohort consisted of 1418 patients who had a complete colonoscopy during which one or more adenomas of the colon or rectum were removed. The patients subsequently underwent periodic colonoscopy during an average follow-up of 5.9 years, and the incidence of colorectal cancer was ascertained. The incidence rate of colorectal cancer was compared with that in three reference groups, including two cohorts in which colonic polyps were not removed and one general-population registry, after adjustment for sex, age, and polyp size. RESULTS: Ninety-seven percent of the patients were followed clinically for a total of 8401 person-years, and 80 percent returned for one or more of their scheduled colonoscopies. Five asymptomatic early-stage colorectal cancers (malignant polyps) were detected by colonoscopy (three at three years, one at six years, and one at seven years). No symptomatic cancers were detected. The numbers of colorectal cancers expected on the basis of the rates in the three reference groups were 48.3, 43.4, and 20.7, for reductions in the incidence of colorectal cancer of 90, 88, and 76 percent, respectively (P < 0.001). CONCLUSIONS: Colonoscopic polypectomy resulted in a lower-than-expected incidence of colorectal cancer. These results support the view that colorectal adenomas progress to adenocarcinomas, as well as the current practice of searching for and removing adenomatous polyps to prevent colorectal cancer. PMID- 8247073 TI - Molecular diagnosis of familial adenomatous polyposis. AB - BACKGROUND: Familial adenomatous polyposis is an inherited disease characterized by multiple colorectal tumors. The diagnosis has classically been based on the detection of multiple colorectal adenomas. The recent identification of germline mutations of the APC gene in patients with familial adenomatous polyposis makes presymptomatic molecular diagnosis possible, but the widespread distribution of the many mutations within this very large gene have heretofore made the search for such mutations impractical. We describe a novel approach that allows molecular genetic diagnosis in the majority of patients with the disease. METHODS: We screened 62 unrelated patients from the Johns Hopkins Familial Adenomatous Polyposis Registry for germline APC mutations. Primary screening was accomplished by analysis of protein synthesized in vitro from surrogate APC genes. In addition, the relative amount of transcript from each APC allele was determined with an allele-specific--expression assay. RESULTS: The protein assay revealed truncated protein in 51 of the 62 patients (82 percent). In 3 of the 11 remaining patients, the allele-specific--expression assay revealed significantly reduced expression of one allele of the APC gene. The use of these two assays in combination successfully identified germline APC mutations in 87 percent of the 62 patients. CONCLUSIONS: The protein and allele-specific--expression assays provide a practical and sensitive method for molecular diagnosis of familial adenomatous polyposis. This approach will facilitate care, allowing routine testing of subjects at risk and genetic confirmation of spontaneous mutations. PMID- 8247074 TI - Insulin resistance and insulin secretory dysfunction as precursors of non-insulin dependent diabetes mellitus. Prospective studies of Pima Indians. AB - BACKGROUND: The relative roles of obesity, insulin resistance, insulin secretory dysfunction, and excess hepatic glucose production in the development of non insulin-dependent diabetes mellitus (NIDDM) are controversial. We conducted a prospective study to determine which of these factors predicted the development of the disease in a group of Pima Indians. METHODS: A body-composition assessment, oral and intravenous glucose-tolerance tests, and a hyperinsulinemic- euglycemic clamp study were performed in 200 non-diabetic Pima Indians (87 women and 113 men; mean [+/- SD] age, 26 +/- 6 years). The subjects were followed yearly thereafter for an average of 5.3 years. RESULTS: Diabetes developed in 38 subjects during follow-up. Obesity, insulin resistance (independent of obesity), and low acute plasma insulin response to intravenous glucose (with the degree of obesity and insulin resistance taken into account) were predictors of NIDDM: The six-year cumulative incidence of NIDDM was 39 percent in persons with values below the median for both insulin action and acute insulin response, 27 percent in those with values below the median for insulin action but above that for acute insulin response, 13 percent in those with values above the median for insulin action and below that for acute insulin response, and 0 in those with values originally above the median for both characteristics. CONCLUSIONS: Insulin resistance is a major risk factor for the development of NIDDM: A low acute insulin response to glucose is an additional but weaker risk factor. PMID- 8247075 TI - A controlled trial of high-dose intravenous immune globulin infusions as treatment for dermatomyositis. AB - BACKGROUND: Dermatomyositis is a clinically distinct myopathy characterized by rash and a complement-mediated microangiopathy that results in the destruction of muscle fibers. In some patients the condition becomes resistant to therapy and causes severe physical disabilities. METHODS: We conducted a double-blind, placebo-controlled study of 15 patients (age, 18 to 55 years) with biopsy-proved, treatment-resistant dermatomyositis. The patients continued to receive prednisone (mean daily dose, 25 mg) and were randomly assigned to receive one infusion of immune globulin (2 g per kilogram of body weight) or placebo per month for three months, with the option of crossing over to the alternative therapy for three more months. Clinical response was gauged by assessing muscle strength, neuromuscular symptoms, and changes in the rash. Changes in immune-mediated muscle abnormalities were determined by repeated muscle biopsies. RESULTS: The eight patients assigned to immune globulin had a significant improvement in sores of muscle strength (P < 0.018) and neuromuscular symptoms (P < 0.035), whereas the seven patients assigned to placebo did not. With crossovers a total of 12 patients received immune globulin. Of these, nine with severe disabilities had a major improvement to nearly normal function. Their mean muscle-strength scores increased from 74.5 to 84.7, and their neuromuscular symptoms improved. Two of the other three patients had mild improvement, and one had no change in his condition. Of 11 placebo-treated patients, none had a major improvement, 3 had mild improvement, 3 had no change in their condition, and 5 had worsening of their condition. Repeated biopsies in five patients of muscles whose strength improved to almost normal showed an increase in muscle-fiber diameter (P < 0.04), an increase in the number and a decrease in the diameter of capillaries (P < 0.01), resolution of complement deposits on capillaries, and a reduction in the expression of intercellular adhesion molecule 1 and major-histocompatibility complex class I antigens. CONCLUSIONS: High-dose intravenous immune globulin is a safe and effective treatment for refractory dermatomyositis. PMID- 8247076 TI - Images in clinical medicine. Mycosis fungoides. PMID- 8247077 TI - Entrapment neuropathies of the upper extremities. AB - The three syndromes described in this article were chosen from about a dozen nerve-compression disorders that affect the arm from scapula to digits. They illustrate the range of problems encountered. None of the other syndromes are as common, and in some instances (pronator syndrome and anterior interosseous syndrome), localized inflammation is at least as common a cause of neuropathy as is compression. Some syndromes of nerve compression have important implications for musicians, athletes, and those who place stress on the upper limbs through heavy or repeated use. Well-designed prospective trials are clearly needed to expand our knowledge of the prevention and treatment of the entrapment neuropathies. Much more needs to be known about the long-term natural history of these disorders, since prevention is usually possible. PMID- 8247078 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 52-1993. A 17-year-old girl with massive hemoptysis and acute oliguric renal failure. PMID- 8247079 TI - From the molecule to public health. PMID- 8247080 TI - Diazepam to prevent febrile seizures. PMID- 8247081 TI - Diazepam to prevent febrile seizures. PMID- 8247082 TI - Diazepam to prevent febrile seizures. PMID- 8247083 TI - Diazepam to prevent febrile seizures. PMID- 8247084 TI - More on HTLV tax and mycosis fungoides. PMID- 8247085 TI - More on HTLV tax and mycosis fungoides. PMID- 8247086 TI - Questionable reliability of the polymerase chain reaction in the detection of Mycobacterium tuberculosis. PMID- 8247087 TI - Screening for hemochromatosis. PMID- 8247088 TI - Screening for hemochromatosis. PMID- 8247089 TI - Effect of tretinoin on collagen formation in photodamaged skin. PMID- 8247090 TI - Health insurance coverage and clinical outcomes in women with breast cancer. PMID- 8247091 TI - Images in clinical medicine: colocolic fistula. PMID- 8247092 TI - Clinical problem-solving: leptospirosis. PMID- 8247093 TI - Clinical problem-solving: leptospirosis. PMID- 8247094 TI - A preliminary survey of cycloheximide-resistant airborne fungi in Turin, Italy. AB - Numbers and type of the cycloheximide-resistant part of the aerially transmitted mycoflora of Turin were studied. Samples from three areas characterized by differing human usage were taken in the first week of March. During each sampling, 12 m3 of air were aspirated, using an one-stage volumetric sieve sampler. Fifty-two mesophilic species and eight thermotolerant were isolated. Propagule load varied from 2.92 to 120.31 cfu m-3. The following species appear not to have been reported previously from air samples: Ascotricha bosei, Blastobotrys navarrensis, Cryptendoxyla hypophloia, Chrysosporium an. gymnoascus demonbreunii, Ophiostoma piceae, Penicillium vulpinum, Phialophora mustea, Rhinocladiella pedrosoi, Scopulariopsis croci, S. komngii and Sesquiallium candelabrum. A significant number of potential opportunistic pathogens was isolated. PMID- 8247095 TI - Influence of different levels of ammonium concentrations on cell growth, RNA and protein production by Candida albicans. AB - Candida albicans starved cells were incubated in minimal synthetic liquid media containing different concentrations of ammonium sulphate (0.00, 0.02, 0.05, 0.10, 0.03, 0.50 g/L). Culture growth was monitored by measuring daily the optical density and by evaluating RNA and protein cellular content after 48 and 96 hours from the inoculum. The environmental availability of ammonium ion influenced the biomass production, that was maximum when its concentration was 0.10 and 0.30 g/L. In addition, an effect on cell duplication time was observed, this was particularly evident when the (NH4)2SO4 concentration was 0.10 g/L. The protein content increased in relation to the increase of ammonium ion availability, with a peak in correspondence to 0.30 g/L and a drop when the greatest concentrations were employed. RNA production was inversely proportional in respect to protein production. The optimal range of ammonium sulphate concentration for C. albicans growth was 0.10-0.30 g/L; over these concentrations there was an inhibitory effect. The rate of the protein and RNA syntheses seems to indicate the growth phase and the nitrogen nutritional conditions of the cultures, respectively. PMID- 8247097 TI - Vitamin B6 and arteriosclerosis. AB - In 1949, Rinehart and Greenberg reported that marked arteriosclerosis occurs in vitamin-B6-deficient monkeys. The present study investigates the relationship between vitamin B6 and arteriosclerosis and summarizes the results. I found that thrombogenesis, disorder of collagen metabolism and production of free radicals may be the processes that cause arteriosclerosis in human and experimental animals with vitamin B6 deficiency. PMID- 8247096 TI - Treatment of systemic candidiasis in a neutropenic murine model using immunoglobulin G bearing liposomal amphotericin B. AB - Efficacy of immunoglobulin G (IgG) bearing liposomal amphotericin B (LAMB-IgG), liposomal amphotericin B without IgG (LAMB) or free amphotericin B (fAMB/Fungizone) was investigated in the treatment of systemic candidiasis in a neutropenic mouse model. Treatment with a single dose (0.6 or 0.9 mg amphotericin B per kg body weight) of LAMB-IgG resulted in a significant increase in the survival rate of neutropenic mice infected with 3 x 10(5) cfu of Candida albicans compared to untreated controls, mice injected with IgG, or liposome alone. Survival was also better in neutropenic mice treated with LAMB-IgG than in neutropenic mice treated with the same dose of LAMB or fAMB. Moreover, 65% of all mice survived the infection after treatment with a single dose of 0.6 mg AMB of the LAMB-IgG formulation. Quantitative culture counts of organs showed that both fAMB and LAMB-IgG formulations even at a dose of 0.3 mg AMB/kg, cleared C. albicans from the spleens, livers, and lungs but not from the kidneys. However, a decreased number of C. albicans cells was recovered from the kidneys of mice that survived the infection. Results of the study suggest that LAMB-IgG is more effective than LAMB or fAMB in the therapy of disseminated candidiasis in neutropenic mice. PMID- 8247098 TI - Remission of myasthenia gravis: clinical, electrophysiological and immunological studies. AB - The prognosis of 142 patients with myasthenia gravis (MG) was clinically investigated. Forty-nine (35%) had clinical remission (CR) and 23 (16%) good improvement (GI), while 70 (49%) remained in poor condition. Favorable clinical factors for CR were the onset of MG before the age of 20 years, a pre-thymectomy period of less than one year, and a post-thymectomy period of six years or more. Single-fiber electromyography (SFEMG) showed abnormal jitter in nine (47%) of the 19 CR patients, while abnormal jitter was shown in 13 (81%) of the 16 GI patients. Abnormal jitter in CR patients was correlated with the following clinical factors: complication with the thymoma, a period of three or more years from thymectomy to remission, and a remission period of less than six years. An anti-acetylcholine receptor (anti-AChR) antibody was positive in 12 CR patients (63%) as well as in 13 GI patients (81%). Based upon these facts, we point out that true remission seldom occurs in MG patients, and that there exist clinical features that may favorably induce clinical remission. We would like to postulate that electrophysiological and immunological follow-up is indispensable even in CR patients to predict recurrence. PMID- 8247099 TI - Cerebellar hypoplasia in the hyperbilirubinemic Gunn rat: morphological aspects. AB - Gunn rats, a mutant strain of rats, suffer from autosomal recessive hyperbilirubinemia. The homozygotes (j/j) develop jaundice soon after birth and often exhibit kernicterus and cerebellar hypoplasia that are due to bilirubin. Therefore, j/j Gunn rats have been used as an animal model of bilirubin encephalopathy, as well as of neonatal hyperbilirubinemia. In this review, we discuss morphological aspects of the cerebellar hypoplasia that is due to bilirubin and describe the relationship between plasma bilirubin levels and cerebellar hypoplasia, as well as the pathogenesis of cerebellar hypoplasia, including abnormal histogenesis of the cerebellar cortex, abnormalities associated with Purkinje cells and abnormal synaptogenesis in j/j Gunn rats. PMID- 8247100 TI - Hepatic copper accumulation in patients with primary biliary cirrhosis. AB - Liver biopsy specimens from 18 patients with primary biliary cirrhosis were examined histochemically and by energy-dispersive x-ray microanalysis. Using two indices, we classified hepatic copper accumulation into three stages based on the Cu x-ray intensity of cuproproteins that had accumulated in hepatocyte lysosomes and on the binding ratio of postulated copper transfer proteins between the cytosol and lysosomes. Eight patients were in stage 1 with an initial accumulation of lysosomal cuproproteins, mediated by transfer proteins not saturated with copper. Two patients were in stage 2, in which transfer proteins were saturated with copper. The first two stages gave negative results for histochemical copper. The remaining eight patients were in stage 3, in which copper accumulation detected by histochemical included transfer proteins saturated with copper and large amounts of lysosomal cuproproteins. Five patients (one each in stages 1 and 2, and three in stage 3) underwent a second liver biopsy after treatment with 600 mg of ursodeoxycholic acid daily for 14 to 39 months. Results of blood chemistry tests improved, but liver histologic findings and copper accumulation were unchanged in all five patients. It seems likely that ursodeoxycholic acid does not affect the copper accumulation in hepatocyte lysosomes that reflects the state of cholestasis in patients with primary biliary cirrhosis. PMID- 8247101 TI - Expression of proto-oncogenes and tumor suppressor genes in in vitro cell lines derived from a thymus, thymoma, and malignant thymoma of rats. AB - To analyze the processes of the development of thymoma and malignant thymoma from normal thymic epithelial cells, we have examined the expression of 15 proto oncogenes and tumor suppressor genes in seven in vitro epithelial cell lines established from a normal thymus (TuD1-1, TuD1-3, and TuD1-5), thymoma (TaD1-3 and TaD1-8), and malignant thymoma (MTHC-1 and MTHC-3) of rats. Northern blot analysis indicated that most of these genes examined were transcribed at similar levels. However, higher levels of transcription of epidermal growth factor receptor (EGFR) were observed in TuD1-1, TuD1-3, TuD1-5, TaD1-3, and TaD1-8 cells than in MTHC-1 and MTHC-3 cells. Conversely, four of the former five cell lines showed no TGF-beta transcription while the latter two cell lines had high levels of its expression. In addition, c-fos proto-oncogene was highly expressed in TuD1 5 cells, which showed the fastest growth rate among the seven cell lines. These results denote that some molecular changes in the regulation of gene expression occurred in the processes of malignant transformation of thymic epithelial cells. PMID- 8247102 TI - Intrahepatocellular localization of copper in Wilson's disease. AB - It is well known that excess copper plays a role in the pathogenesis of Wilson's disease; however, the hepatic copper contents, determined either histochemically or biochemically, are not always correlated with the activity of Wilson's disease. To better understand copper-induced cytotoxicity, intrahepatocellular localization of copper was studied in five patients with Wilson's disease. The liver specimens were obtained by biopsy to confirm the clinical diagnosis of Wilson's disease before treatment. Neither hepatic copper content nor histochemical copper deposits were correlated with any of the biochemical indices studied. Energy-dispersion X-ray microanalysis was done on the nuclei, lysosomes and lysosome-free cytoplasm of hepatocytes. Lysosomes had the highest Cu X-ray intensity but no correlation was shown between the lysosomal copper content and biochemical indices. The nucleus/lysosome-free cytoplasm ratio of the copper content was correlated with the serum levels of aminotransferases. These results suggest that the copper increasing gradient from the lysosome-free cytoplasm to the nucleus is associated with hepatocyte necrosis, and probably causes irreversible nuclear damage. PMID- 8247103 TI - Measles and measles vaccine in Japan. AB - Before the introduction of measles vaccine in 1971, measles was a common and well recognized disease in Japanese children. Seroepidemiological studies conducted before the general use of the vaccine disclosed that measles epidemics differed from community to community; in rural areas, epidemics appeared at intervals of several years with the accumulation of susceptibles, and in urban areas, measles was endemic affecting younger children. The measles vaccines developed in Japan showed excellent safety and efficacy in the clinical trials for general use. Since 1978 measles vaccines have been routinely given to children aged 12 months or older, and incidence of measles decreased dramatically. However, the vaccine acceptance rate has been only as high as 85% to 90% and small-scale outbreaks of measles have been observed periodically among unimmunized children: Measles is far from being eradicated in Japan. This is primarily due to the insufficient vaccine acceptance rate, and more efforts towards mobilization of parents to have their children vaccinated are now in progress. PMID- 8247104 TI - Considerations for standard chest radiography: the long film-focus distance technique. AB - The possibility of using long film-focus distance radiography as a new screening method for lung cancer diagnosis was investigated. Long-film focus distance radiography produced exceptionally superior image sharpness suitable for use as clinical photographs, at a distance of about FFD (Film-Focus Distance) 20 m. The effective energy was increased by narrowing the radiation spectrum via an air filter effect, which may prove useful in reducing radiation exposure dosage. Questionnaire results indicate good visualization of cardiac posterior and trachea shadow regions in the diaphragm. Further study is required regarding methods for stabilizing patient position and synchronization with heart beat and respiration to improve reproducibility. PMID- 8247105 TI - Cisplatin, etoposide, and vincristine combination chemotherapy in the treatment of non-small cell lung cancer. AB - Thirty patients with previously untreated, inoperable non-small cell lung cancer (NSCLC) were treated with cisplatin, etoposide and vincristine. Among twenty-nine evaluable patients, eight patients achieved partial response and the overall response rate was 28%. No patient achieved a complete response. The median survival time was 51 weeks. Myelosuppression was the dose-limiting toxicity. Four patients had a leukocyte nadir of less than 1000/mm3, and one died from severe myelosuppression and sepsis. The other toxicities were nausea/vomiting, peripheral neuropathy, and alopecia. We conclude that cisplatin, etoposide, and vincristine combination chemotherapy offers moderate activity for inoperable non small cell lung cancer. PMID- 8247106 TI - Ultrastructural and morphometrical studies on the reticular framework and reticular fibers in the reticuloendothelial system of rats. AB - The reticular framework and reticular fibers in the thymus, cervical lymph node, spleen and bone marrow of Wistar rats were studied by transmission electron microscopy and morphometrical method. The reticular framework was usually observed in these organs as a common structure that consisted mainly of the slender cytoplasmic processes of the fibroblastic reticular cells interconnected with tight junction. Ultrastructurally, the reticular fibers were a mixture of collagen fibrils and amorphous materials, and were almost completely ensheathed by the cytoplasm of fibroblastic reticular cells. Such characteristic structure of the reticular fibers was found not only in the thymus, lymph node and spleen, but also in the bone marrow where it has not been clearly demonstrated previously. Morphometrical analysis revealed that the content of collagen fibrils in the reticular fiber in the lymphoid tissues (the thymus, lymph node and splenic white pulp) was much greater than that in the hematopoietic tissues (the bone marrow and splenic red pulp). Based on these evidences, it was reasonably considered that the reticular framework and reticular fiber ensheathed by the cytoplasm of the fibroblastic reticular cells were the most representative common structure in the reticuloendothelial system (RES) and played some important roles in the functions of RES. PMID- 8247107 TI - Quantitative analysis of species identification tests of bloodstains using anti human serum. AB - Immunological reactions of bloodstains of human and animals to anti-human serum were analysed quantitatively to determine which technique is the most effective to differentiate human bloodstains from those of animals. The lower threshold of counter-electrophoresis was 10-25 micrograms protein/ml and increased only slightly with increasing age of the bloodstains, while that of the ring test and immunodiffusion decreased greatly. In counter-electrophoresis, absorption of anti human serum by Japanese monkey serum resulted in a marked decrease in the cross reaction with animals, and still showed no change in the lower threshold for human samples. The present results show that counter-electrophoresis is especially useful for the identification of human bloodstains. PMID- 8247108 TI - The relationship of high-density lipoprotein cholesterol to obesity, drinking and smoking habits. AB - Subjects were a total of 726 Japanese male transport service workers between the ages of 35 and 50. They were analyzed for obesity and drinking or smoking habits in relation to their HDL-cholesterol level and total cholesterol concentration. The cases were 100 low HDL-cholesterol (less than 40 mg/dl) and 121 high total cholesterol (greater than 230 mg/dl) subjects. We used a multiple logistic regression analysis. As for abnormally high total cholesterol, it was determined that the odds ratios at 40, 45, and 50 years of age were 1.95, 2.23, and 2.65, respectively, when 35-year-olds were considered the baseline. Abnormally high total cholesterol showed an odds ratio of 0.27 among thin subjects with a Body Mass Index (BMI) less than 21.0, when subjects with a standard BMI were calculated as the baseline. With reference to low HDL-cholesterol, the odds ratio for lean subjects was 0.30 against those who had standard habitus, while subjects with a BMI of more than 25.1 had an odds ratio of more than double. Compared with nondrinkers, low HDL-cholesterol exhibited odds ratios of 0.16 and 0.23, respectively, for moderate and heavy drinkers. In cases of low HDL-cholesterol, the odds ratios for mild and heavy smokers were 1.91 and 2.27, respectively, versus nonsmokers. PMID- 8247109 TI - Microassay of sperm concentration in the rat epididymis by micropuncture technique. AB - Micropuncture samples taken from the rete testis, caput, corpus and cauda epididymidis of adult rats were assayed for intraluminal sperm concentrations. The amount of fluid resorbed from the efferent duct and epididymal lumen was calculated based on the sperm concentration. Epididymal sperm concentrations increased from the rete fluid to the cauda fluid through the caput fluid. Eighty nine percent of the fluid leaving the rete testis is resorbed by the efferent ducts and proximal epididymal tubule, and 96% of the fluid leaving the rete testis is resorbed in the distal cauda epididymidis. Resorption is important for the control of intraluminal fluid by the epididymis. PMID- 8247110 TI - Prosthodontic closure of palatal fistula with osseointegrated implants and onlay bone grafts--case report. AB - A case treated with onlay bone grafting and simultaneous osseointegrated implant insertion is reported. The patient had an oronasal fistula in the center of the premaxillary region due to failed repair of a bilateral cleft palate. Four fixtures were inserted with onlay bone grafting of the maxillary alveolar ridge. Twelve months postsurgery, one fixture was lost as a result of severe bone resorption around the fixture, but three fixtures integrated strongly. We constructed an overdenture stabilized with ball attachments connected to the implants 13 months after the first operation. Speech and masticatory function improved remarkably with closure of the fistula and good fixation of the denture. PMID- 8247111 TI - Cervicofacial actinomycosis: report of two cases. AB - This article presents two cases of actinomycosis. Case 1 was a 39-year-old man who was first seen with the chief complaint of swelling around the left submandibular region. Case 2 was a 40-year-old woman who was first seen with the chief complaint of mass formation around the left buccal area. Both cases were initially diagnosed as malignant tumors and were later histopathologically interpreted as actinomycosis because of the presence of sulfur granules. PMID- 8247112 TI - Cuproproteins of hepatocyte lysosomes in normal and fatty liver. AB - The pathophysiological significance of cuproproteins in hepatocyte lysosomes was studied in 7 patients with structurally normal livers and 13 patients with fatty livers by energy-dispersion X-ray microanalysis. In most patients, in both groups, copper and sulfur coexisted in hepatocyte lysosomes. The correlation between copper and sulfur indicated that copper transfer between the cytosol and lysosomes was mediated by a cuproprotein. The copper indices of the lysosomal proteins and transfer proteins were high in some fatty livers. These observations suggest that cuproproteins in hepatocytes are not necessarily pathological, but are present in greater amounts in some fatty livers, probably because of impaired bile drainage. PMID- 8247113 TI - Solitary intraosseous neurofibroma of the mandible: report of a case. AB - A case of solitary intraosseous neurofibroma of the mandible is reported. The tumor was located in the body of the right mandible. Segmental mandibulectomy including the surrounding tissue was performed and the mandible was reconstructed with the free scapular osteocutaneous flap. From histological examination of the resected mandible, typical findings of a tumor invading the adjacent bone along the mandibular nerve were observed. Immunohistochemical examination of the tumor showed positive staining of the tumor cells for anti-S100 protein antibodies. Numerous nerve-like fibers were also noted by Bodian stain. PMID- 8247114 TI - Court backs critic of Italian jobs 'carve-up'. PMID- 8247115 TI - WHO widens focus of AIDS research. PMID- 8247116 TI - Trial deaths prompt tighter rules. PMID- 8247117 TI - Evolution and religion. PMID- 8247118 TI - Evolution and religion. PMID- 8247119 TI - DNA repair. The nick in time. PMID- 8247120 TI - Genomic imprinting. Silence of the genes. PMID- 8247121 TI - Developmental biology. A no-wing situation. PMID- 8247122 TI - Hormone signal response system. PMID- 8247123 TI - Death and c-fos. PMID- 8247124 TI - Water in life. PMID- 8247125 TI - The yeast DOA4 gene encodes a deubiquitinating enzyme related to a product of the human tre-2 oncogene. AB - Modification of specific intracellular proteins by ubiquitin targets them for degradation. We describe a yeast enzyme, Doa4, that is integral to the degradation of ubiquitinated proteins and is required in diverse physiological processes. Doa4 appears to function late in the proteolytic pathway by cleaving ubiquitin from substrate remnants still bound to protease. The human tre-2 oncogene encodes a deubiquitinating enzyme similar to Doa4, indicating a role for the ubiquitin system in mammalian growth control. PMID- 8247126 TI - Self-assembling organic nanotubes based on a cyclic peptide architecture. AB - Hollow tubular structures of molecular dimensions may offer a variety of applications in chemistry, biochemistry and materials science. Concentric carbon nanotubes have attracted a great deal of attention, while the three-dimensional tubular pore structures of molecular sieves have long been exploited industrially. Nanoscale tubes based on organic materials have also been reported previously. Here we report the design, synthesis and characterization of a new class of organic nanotubes based on rationally designed cyclic polypeptides. When protonated, these compounds crystallize into tubular structures hundreds of nanometres long, with internal diameters of 7-8 A. Support for the proposed tubular structures is provided by electron microscopy, electron diffraction, Fourier-transform infrared spectroscopy and molecular modelling. These tubes are open-ended, with uniform shape and internal diameter. We anticipate that they may have possible applications in inclusion chemistry, catalysis, molecular electronics and molecular separation technology. PMID- 8247127 TI - The retinoid ligand 4-oxo-retinoic acid is a highly active modulator of positional specification. AB - Retinoids (vitamin A and its metabolites) are suspected of regulating diverse aspects of growth, differentiation, and patterning during embryogenesis, but many questions remain about the identities and functions of the endogenous active retinoids involved. The pleiotropic effects of retinoids may be explained by the existence of complex signal transduction pathways involving diverse nuclear receptors of the retinoic acid receptor (RAR) and retinoid X receptor (RXR) families, and at least two types of cellular retinoic acid binding proteins (CRABP-I and -II). The different RARs, RXRs, and CRABPs have different expression patterns during vertebrate embryogenesis, suggesting that they each have particular functions. Another level at which fine tuning of retinoid action could occur is the metabolism of vitamin A to active metabolites, which may include all trans-retinoic acid, all-trans-3,4-didehydroretinoic acid, 9-cis-retinoic acid, and 14-hydroxy-4,14-retroretinol. Formation of the metabolite all-trans-4-oxo retinoic acid from retinoic acid was considered to be an inactivation pathway during growth and differentiation. We report here that, in contrast, 4-oxo retinoic acid is a highly active metabolite which can modulate positional specification in early embryos. We also show that this retinoid binds avidly to and activates RAR beta, and that it is available in early embryos. The different activities of 4-oxo-retinoic acid and retinoic acid in modulating positional specification on the one hand, and growth and differentiation on the other, interest us in the possibility that specific retinoid ligands regulate different physiological processes in vivo. PMID- 8247128 TI - Dopamine D1 receptors facilitate transmitter release. AB - A physiological role for the dopamine D1 receptor has been difficult to define, particularly because of its complex pre- and postsynaptic localization in brain areas such as the striatum. In the midbrain, however, D1 receptors are selectively localized to the terminals of GABA (gamma-aminobutyric acid) containing afferents. We have studied the actions of these D1 receptors on evoked GABA synaptic potentials recorded intracellularly from dopamine neurons in the ventral tegmental area (VTA). We report here that dopamine augmented GABAB inhibitory postsynaptic potentials (i.p.s.ps) in the presence of D2 receptor antagonists. This effect was mimicked by the D1 agonists SKF38393 and SKF82958 and blocked by the D1 antagonists SCH23390 and cis-flupenthixol. No modulation of the GABAA synaptic potential was observed. The postsynaptic actions of the GABAB agonist, baclofen, were unaffected by SKF38393, SCH23390 or cis-flupenthixol, confirming a presynaptic locus of D1 action. Additionally, D1 antagonists reduced the amplitude of the GABAB i.p.s.p. in the absence of D1 agonists. We conclude that dopamine acts tonically at presynaptic D1 receptors on the terminals of afferent GABA neurons to facilitate selectively GABAB-mediated neurotransmission in the midbrain. PMID- 8247129 TI - Synaptic vesicle fusion complex contains unc-18 homologue bound to syntaxin. AB - Three synaptic proteins, syntaxin, SNAP-25 and synaptobrevin, were recently identified as targets of clostridial neurotoxins that irreversibly inhibit synaptic vesicle fusion. Experiments searching for membrane receptors for N ethylmaleimide-sensitive fusion protein (NSF), which has an important role in membrane fusion, revealed an ATP-dependent interaction of the same three synaptic proteins with NSF and its soluble attachment proteins. Thus, two independent approaches identify syntaxin, synaptobrevin and SNAP-25 as components of the synaptic vesicle fusion machinery, but their mode of action is unclear. We have now discovered a brain protein of relative molecular mass 67,000 (67K) which binds stably to syntaxin. Amino-acid sequencing and complementary DNA cloning revealed that the 67K protein is encoded by the mammalian homologue of the Caenorhabditis elegans gene unc-18. In C. elegans, unc-18 belongs to a group of genes defined by mutations with a paralytic phenotype and accumulations of acetylcholine, suggesting a defect in neurotransmitter release. The binding of the mammalian homologue of unc-18 (Munc-18) to syntaxin requires the N terminus of syntaxin whereas that of SNAP-25 involves a more C-terminal sequence. Our data suggest that Munc-18 is a previously unidentified essential component of the synaptic vesicle fusion protein complex. PMID- 8247130 TI - GTP binding and hydrolysis by the signal recognition particle during initiation of protein translocation. AB - The signal recognition particle (SRP) consists of one RNA and six protein subunits. The N-terminal domain of the 54K subunit contains a putative GTP binding site, whereas the C-terminal domain binds signal sequences and SRP RNA. Binding of SRP to the signal sequence as it emerges from the ribosome creates a cytosolic targeting complex containing the nascent polypeptide chain, the translating ribosome, and SRP. This complex is directed to the endoplasmic reticulum membrane as a result of its interaction with the SRP receptor, a membrane protein composed of two subunits, SR alpha and SR beta, each of which also contains a GTP-binding domain. In the presence of GTP, SRP receptor binding to SRP causes the latter to dissociate from both the signal sequence and the ribosome. GTP is then hydrolysed so that SRP can be released from the SRP receptor and returned to the cytosol. Here we show that the 54K subunit (M(r) 54,000) of SRP (SRP54) is a GTP-binding protein stabilized in a nucleotide-free state by signal sequences, and that the SRP receptor both increases the affinity of SRP54 for GTP and activates its GTPase. We propose that nucleotide-mediated conformational changes in SRP54 regulate the release of signal sequences and the docking of ribosomes at the endoplasmic reticulum. PMID- 8247131 TI - Defective mitosis due to a mutation in the gene for a fission yeast 26S protease subunit. AB - We have isolated a mutant, mts2, in the fission yeast Schizosaccharomyces pombe which is defective in chromosome segregation. The predicted amino-acid sequence of the cloned mts2+ gene product is 75% identical to the S4 subunit of the human 26S ATP/ubiquitin-dependent protease. The human S4 subunit complementary DNA expressed from an S. pombe expression plasmid can rescue an S. pombe mts2 gene disruption. Both observations demonstrate that the mts2+ gene is the S. pombe homologue of the human S4 subunit. In addition, we provide genetic evidence for a physical interaction between the S4 and the related S7 subunit in the 26S multiprotein protease. We show that polyubiquitin-conjugated proteins accumulate in the mts2 mutant at the restrictive temperature, demonstrating that the mutant has an in vivo defect in the ubiquitin-dependent proteolysis pathway. Finally, the phenotype for the mts2 mutant indicates that protein degradation by the 26S protease is essential not for entry into but for the completion of mitosis. PMID- 8247132 TI - S. cerevisiae 26S protease mutants arrest cell division in G2/metaphase. AB - We isolated two mutants from the yeast Saccharomyces cerevisiae, cim3-1 and cim5 1, that arrest cell division in G2/metaphase at 37 degrees C. CIM3 (identical to SUG1; ref. 1) and CIM5 are similar to each other and are members of a family of putative ATPases that have been proposed to be 26S protease subunits. We show here that CIM5 is the functional yeast homologue of the human MSS1 protein and that homologues of CIM3 and CIM5 are present in a highly purified preparation of the Drosophila 26S protease. The short-lived ubiquitin-proline-beta-galactosidase fusion protein is stabilized in cim mutants, but Leu-beta-galactosidase is not. The CLB2 and CLB3 cyclins also accumulate in the cim mutants. Thus the 26S protease is required in vivo for the degradation of ubiquitinated substrates and for anaphase chromosome separation. PMID- 8247133 TI - Role for DNA methylation in genomic imprinting. AB - The paternal and maternal genomes are not equivalent and both are required for mammalian development. The difference between the parental genomes is believed to be due to gamete-specific differential modification, a process known as genomic imprinting. The study of transgene methylation has shown that methylation patterns can be inherited in a parent-of-origin-specific manner, suggesting that DNA methylation may play a role in genomic imprinting. The functional significance of DNA methylation in genomic imprinting was strengthened by the recent finding that CpG islands (or sites) in three imprinted genes, H19, insulin like growth factor 2 (Igf-2), and Igf-2 receptor (Igf-2r), are differentially methylated depending on their parental origin. We have examined the expression of these three imprinted genes in mutant mice that are deficient in DNA methyltransferase activity. We report here that expression of all three genes was affected in mutant embryos: the normally silent paternal allele of the H19 gene was activated, whereas the normally active paternal allele of the Igf-2 gene and the active maternal allele of the Igf-2r gene were repressed. Our results demonstrate that a normal level of DNA methylation is required for controlling differential expression of the paternal and maternal alleles of imprinted genes. PMID- 8247134 TI - Yeast excision repair gene RAD2 encodes a single-stranded DNA endonuclease. AB - In eukaryotes nucleotide excision repair of DNA damaged by ultraviolet radiation requires several gene products; defects in this process result in the cancer prone syndrome xeroderma pigmentosum (XP) in humans. The RAD2 gene is one of at least seven genes indispensable for excision repair in the yeast Saccharomyces cerevisiae, and its encoded protein shares remarkable homology with the XP group G gene product. Here we overproduce the RAD2-encoded protein in S. cerevisiae, purify it to near homogeneity, and show that RAD2 protein in the presence of magnesium degrades circular single-stranded DNA. The RAD2 endonuclease is specific for single-stranded DNA as it does not act on double-stranded DNA. Given the absolute requirement for RAD2 in the incision step of excision repair, our findings directly implicate RAD2 protein and its human homologue XPG protein as a catalytic component that incises the damaged DNA strand during excision repair. Furthermore, our results indicate that eukaryotes probably employ two distinct endonuclease activities to mediate the dual incision at the damage site. PMID- 8247135 TI - UK clinical geneticists ask for ban on the patenting of human genes. PMID- 8247136 TI - EC pays up for human frontiers program. PMID- 8247137 TI - Top US panel will map out fresh attack on AIDS. PMID- 8247138 TI - Effects on health of mustard gas. PMID- 8247139 TI - Effects on health of mustard gas. PMID- 8247140 TI - Fidia and neuroscience. PMID- 8247141 TI - Fidia and neuroscience. PMID- 8247142 TI - Fidia and neuroscience. PMID- 8247143 TI - Gerontology. Methuselah among nematodes. PMID- 8247144 TI - Evolutionary physiology. Quantitative design of life. PMID- 8247145 TI - Severo Ochoa (1905-93) PMID- 8247146 TI - Molecular neurobiology. The chimaeras speak again. PMID- 8247147 TI - Fatal attraction. PMID- 8247148 TI - Dating hominid remains. PMID- 8247149 TI - Quantitative PCR. PMID- 8247150 TI - The human IgE network. AB - IgE and its receptors are believed to have evolved as a mechanism to protect mammals against parasites. But other and intrinsically innocuous antigens can subvert this system to provoke an allergic response. For human populations in industrialized countries, allergy and asthma now represent a far greater threat than parasitic infection, and the main impetus for current studies of the IgE system is the hope of understanding and intervening in the aetiology of allergic diseases. PMID- 8247151 TI - Mode-switching of a voltage-gated cation channel is mediated by a protein kinase A-regulated tyrosine phosphatase. AB - Tyrosine kinases and tyrosine phosphatases are abundant in central nervous system tissue, yet the role of these enzymes in the modulation of neuronal excitability is unknown. Patch-clamp studies of an Aplysia voltage-gated cation channel now demonstrate that a tyrosine phosphatase endogenous to excised patches determines both the gating mode of the channel and the response of the channel to protein kinase A. Moreover, a switch in gating modes similar to that triggered by the phosphatase occurs at the onset of a prolonged change in the excitability of Aplysia bag cell neurons. PMID- 8247152 TI - A siderophore from a marine bacterium with an exceptional ferric ion affinity constant. AB - Virtually all microorganisms require iron for growth. The paucity of iron in surface ocean water (approximately 0.02-1.0 nM (refs 1, 2)) has spurred a lively debate concerning iron limitation of primary productivity, yet little is known about the molecular mechanisms used by marine microorganisms to sequester iron. Terrestrial bacteria use a siderophore-mediated ferric uptake system. A siderophore is a low-molecular-mass compound with a high affinity for ferric ion which is secreted by microorganisms is response to low-iron environments; siderophore biosynthesis is regulated by iron levels, with repression by high iron. Although open-ocean marine microorganisms (such as phytoplankton and bacteria) produce siderophores, the nature of these siderophores has not been investigated. We report here the first structure determination, to our knowledge, of the siderophores from an open-ocean bacterium, alterobactin A and B from Alteromonas luteoviolacea. A. luteoviolacea is found in oligotrophic and coastal waters. Alterobactin A has an exceptionally high affinity constant for ferric ion. We suggest that at least some marine microorganisms may have developed higher-affinity iron chelators as part of an efficient iron-uptake mechanism which is more effective than that of their terrestrial counterparts. PMID- 8247153 TI - A C. elegans mutant that lives twice as long as wild type. AB - We have found that mutations in the gene daf-2 can cause fertile, active, adult Caenorhabditis elegans hermaphrodites to live more than twice as long as wild type. This lifespan extension, the largest yet reported in any organism, requires the activity of a second gene, daf-16. Both genes also regulate formation of the dauer larva, a developmentally arrested larval form that is induced by crowding and starvation and is very long-lived. Our findings raise the possibility that the longevity of the dauer is not simply a consequence of its arrested growth, but instead results from a regulated lifespan extension mechanism that can be uncoupled from other aspects of dauer formation. daf-2 and daf-16 provide entry points into understanding how lifespan can be extended. PMID- 8247154 TI - Modulation of the cell cycle contributes to the parcellation of the primate visual cortex. AB - An as-yet unresolved issue in developmental neurobiology is whether the discrete areas that form the mammalian cortex emerge from a uniform cortical plate or whether they are already specified in the germinal zone. A feature of the primate striate cortex is that the number of neurons per unit area is twice that of anywhere else in the cerebral cortex. Here we take advantage of this unique structural feature to investigate whether the extra striate cortical cells are due to increased neuron production during neurogenesis. We labelled precursors undergoing terminal cell division with 3H-thymidine and allowed them to migrate to the cortical plate. Cell counts revealed that their rate of production in the germinal zone of striate cortex is higher than in that given rise to extrastriate cortex. Also, we used 3H-thymidine pulse injections to investigate cell cycle dynamics and found that this phase of increased production of striate cortical cells is associated with changes in the parameters of the cell cycle. These results show that cortical area identity is at least partially determined at the level of the ventricular zone. PMID- 8247155 TI - Neural basis of saccade target selection in frontal eye field during visual search. AB - Conspicuous visual features commonly attract gaze, but how the brain selects targets for eye movements is not known. We investigated target selection in rhesus monkeys performing a visual search task by recording neurons in the frontal eye field, an area known to be responsible for generating purposive eye movements. Neurons with combined visual- and eye movement-related activity were analysed. We found that the initial visual responses to search stimulus arrays were the same whether the target or a distractor was in the response field. We also found that the neural activity evolved to specify target location before the execution of eye movements, ultimately peaking when the target was in the response field and being suppressed when the target was beside but not distant from the response field. These results demonstrate a possible mechanism by which a desired target is fixated and inappropriate eye movements are prevented. PMID- 8247156 TI - Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. AB - Expression of viral genes in transgenic plants is a very effective tool for attenuating plant viral infection. Nevertheless, the lack of generality and risk issues related to the expression of viral genes in plants might limit the exploitation of this strategy. Expression in plants of antibodies against essential viral proteins could provide an alternative approach to engineer viral resistance. Recently, expression of complete or engineered antibodies has been successfully achieved in plants. The engineered single-chain Fv antibody scFv (refs 10, 11) is particularly suitable for expression in plants because of its small size and the lack of assembly requirements. Here we present evidence that constitutive expression in transgenic plants of a scFv antibody, directed against the plant icosahedral tombusvirus artichoke mottled crinkle virus, causes reduction of infection incidence and delay in symptom development. PMID- 8247157 TI - Functional ecdysone receptor is the product of EcR and Ultraspiracle genes. AB - Although the biological activity of the insect moulting hormone ecdysone, is manifested through a hormonally regulated transcriptional cascade associated with chromosomal puffing, a direct association of the receptor with the puff has yet to be established. The cloned ecdysone receptor (EcR) is by itself incapable of high-affinity DNA binding or transcriptional activation. Rather, these activities are dependent on heterodimer formation with Ultraspiracle (USP) the insect homologue of vertebrate retinoid X receptor. Here we report that native EcR and USP are co-localized on ecdysone-responsive loci of polytene chromosomes. Moreover, we show that natural ecdysones selectively promote physical association between EcR and USP, and conversely, that high-affinity hormone binding requires both EcR and USP. Replacement of USP with retinoid X receptor produces heterodimers with distinct pharmacological and functional properties. These results redefine the ecdysone receptor as a dynamic complex whose activity may be altered by combinatorial interactions among subunits and ligand. PMID- 8247158 TI - Chimaeric nicotinic-serotonergic receptor combines distinct ligand binding and channel specificities. AB - The neuronal nicotinic alpha 7 (nAChR) and 5-hydroxytryptamine (5HT3) receptors are ligand-gated ion channels with a homologous topological organization and have activation and desensitization reactions in common. Yet these homo-oligomeric receptors differ in the pharmacology of their binding sites for agonists and competitive antagonists, and in their sensitivity to Ca2+ ions. The alpha 7 channel is highly permeable to Ca2+ ions and external Ca2+ ions potentiate, in an allosteric manner, the permeability response to acetylcholine, as shown for other neuronal nAChRs. The 5HT3 channel, in contrast, is not permeable to Ca2+ ions, but blocked by them. To assign these properties to delimited domains of the primary structure, we constructed several recombinant chimaeric alpha 7-5HT3 receptors. We report here that one of the constructs expresses a functional receptor that contains the serotonergic channel still blocked by Ca2+ ions, but is activated by nicotinic ligands and potentiated by external Ca2+ ions. PMID- 8247159 TI - The crystal structure of a two zinc-finger peptide reveals an extension to the rules for zinc-finger/DNA recognition. AB - The Cys2-His2 zinc-finger is the most widely occurring DNA-binding motif. The first structure of a zinc-finger/DNA complex revealed a fairly simple mechanism for DNA recognition suggesting that the zinc-finger might represent a candidate template for designing proteins to recognize DNA. Residues at three key positions in an alpha-helical 'reading head' play a dominant role in base-recognition and have been targets for mutagenesis experiments aimed at deriving a recognition code. Here we report the structure of a two zinc-finger DNA-binding domain from the protein Tramtrack complexed with DNA. The amino-terminal zinc-finger and its interaction with DNA illustrate several novel features. These include the use of a serine residue, which is semi-conserved and located outside the three key positions, to make a base contact. Its role in base-recognition correlates with a large, local, protein-induced deformation of the DNA helix at a flexible A-T-A sequence and may give insight into previous mutagenesis experiments. It is apparent from this structure that zinc-finger/DNA recognition is more complex than was originally perceived. PMID- 8247160 TI - [Obstruction ileus with leg pain: think of an obturator hernia]. PMID- 8247161 TI - [Adequate adjustment of diabetes mellitus decreases the possibility of diabetic complications]. PMID- 8247162 TI - [Cell and disease. VIII. Pathology of the extracellular matrix]. PMID- 8247163 TI - [Pathogenesis of anaphylactic symptoms following an insect sting]. PMID- 8247164 TI - [Anaphylactic reactions following insect stings: clinical aspects, course and treatment]. PMID- 8247165 TI - [Evaluation of a decade of neonatal screening for congenital hypothyroidism in The Netherlands]. AB - OBJECTIVE: Evaluation of the Dutch screening programme for congenital hypothyroidism (CH). DESIGN: Descriptive. SETTING: Nationwide. METHODS: Data on the screening were obtained from the administration body for vaccination and screening results, laboratories and paediatricians to whom infants with positive screening values were referred, during the period from January 1st, 1981 to December 31st, 1990. RESULTS: Of all live births in the Netherlands, 99.5% (1,797,719) were screened for CH. During the study period, 10,165 children (0.57% of all screened infants) were referred. Of the referred children, 529 had primary CH and 53 had congenital thyrotropin deficiency syndrome (CTDS). The prevalences of primary CH and CTDS are 1:3,400 and 1:25,000, respectively. The sensitivity of the programme with respect to detection of primary CH and CTDS was 99% and 74%, respectively. For all forms of CH combined, the specificity of the programme was 99% and the positive predictive value 6%. The positive predictive value was especially low in the group of infants with low T4 and normal thyrotropin values. One of the goals of the programme is to realise the start of treatment in all patients before they reach the age of three weeks. Before the screening programme came into being, the cumulative proportion of patients treated on the 21st day was 6%. After the beginning of the programme, the proportion increased to 54%. In screened patients with a severe form of CH it is currently 72%. CONCLUSION: The screening programme has made a substantial and important contribution to early and effective identification of patients with CH. A number of measures to decrease the number of false-positive results have been taken and others are at present being investigated. Although patients are now treated much earlier than before the programme started, substantial improvement in this respect is still possible. This can only be achieved by a collective effort of performers of the heel puncture, laboratories, administration body for vaccination and screening results, general practitioners and pediatricians. PMID- 8247166 TI - [Evaluation of various forms of vitamin-K prophylaxis in breastfed infants]. AB - OBJECTIVE: To establish recommendations for prevention of vitamin K deficiency in healthy breastfed infants. DESIGN: Prospective clinical trials of different methods of vitamin K prophylaxis. METHODS: Study of vitamin K1 and proteins induced by vitamin K absence (PIVKA)-II concentrations, and some coagulation factors at the ages of 2, 4, 8 and 12 weeks in healthy breastfed infants with either once 1 mg vitamin K1 orally (n = 165) or intramuscularly (n = 166), or weekly 1 mg orally (n = 48), or daily 25 micrograms orally (n = 58). RESULTS: Despite significantly higher vitamin K1 plasma concentrations after intramuscular administration, there was no difference in activities of coagulation factors VII and X, and PIVKA-II concentrations between oral and intramuscular administration. The two single administrations of 1 mg could not prevent the appearance of PIVKA II after the age of 1 month. When vitamin K was administered as 1 mg per week or 0.025 mg per day, significantly higher concentrations of vitamin K1 were found and no PIVKA-II was detectable. CONCLUSION: A single administration of 1 mg vitamin K1 orally or intramuscularly may not afford complete protection against late vitamin K deficiency in healthy breastfed infants. A regimen of 1 mg per week or 25 micrograms vitamin K1 per day proved to be effective in prevention of vitamin K deficiency and the latter is recommended for breastfed infants during the first three months of life. PMID- 8247167 TI - [HIV prevalence among Surinam and Antillean drug users in Amsterdam]. AB - OBJECTIVE: To determine the HIV prevalence and the frequency of injecting drugs among drug users from Surinam and the Netherlands Antilles in Amsterdam. DESIGN: Descriptive study. SETTING: Amsterdam streets. METHOD: Participants were recruited in 1992 in the street and interviewed about their drug use behaviour, sexual life style and previous HIV test results. Blood or saliva samples were collected for HIV antibody testing. RESULTS: Of 198 participants recruited, 185 (93%) were males. The mean age was 38 years. The mean duration of stay in the Netherlands was 19 years and mean duration of drug use 14 years. Sixty-eight percent of the participants received methadone treatment. Injecting drugs at any time was reported by 29 (15%) participants. Injecting drug users (IDU) more often had a steady partner who also injected or otherwise used drugs. Nearly one quarter reported having had sex with a steady or casual IDU partner. HIV prevalence among IDU was 17% (5/29; 95%-CI 7.4-35%), among heterosexual male non IDU 4.5% (7/156; 95%-CI 2.1-9.1%) and among female non-IDU 9.1% (1/11; 95%-CI 1.3 44%). HIV positive heterosexual non-IDU reported a higher mean number of heterosexual partners in the past 6 months (7.1 versus 1.3), more often had had hepatitis and more often were blood transfusion recipients than HIV negative heterosexual non-IDU. CONCLUSION: HIV prevalence among IDU in this study did not differ from the prevalences found among other IDU in Amsterdam. However, HIV prevalence among heterosexual non-IDU originating from Surinam and the Netherlands Antilles was high when compared with other non-IDU heterosexuals in Amsterdam and may indicate the presence of heterosexual transmission of HIV. Underreporting of risk behaviour or transmission through blood transfusion however, cannot be excluded completely. PMID- 8247168 TI - [A rare cause of hemiparesis: cerebral hydatidosis]. AB - A 6-year-old Turkish boy was brought to the Netherlands by his immigrant father because of hemiparesis. There were no signs of raised intracranial pressure. Cerebral hydatidosis was diagnosed. After surgical removal of the hydatid cyst and postoperative treatment with albendazole the patient recovered. Cerebral hydatidosis is very rare in the Netherlands but should be considered in immigrants from endemic countries. PMID- 8247169 TI - [What is the benefit of intracutaneous injections of bovine collagen and are there side effects with this treatment?]. PMID- 8247170 TI - [No advantage in the addition of dipyridamole or of oral anticoagulants in comparison to low-dose acetylsalicylic acid (50 mg per day) in the prevention of venous transplant occlusion following coronary bypass surgery]. PMID- 8247171 TI - [No advantage in the addition of dipyridamole or of oral anticoagulants in comparison to low-dose acetylsalicylic acid (50 mg per day) in the prevention of venous transplant occlusion following coronary bypass surgery]. PMID- 8247172 TI - [No advantage in the addition of dipyridamole or of oral anticoagulants in comparison to low-dose acetylsalicylic acid (50mg per day) in the prevention of venous transplant occlusion following coronary bypass surgery]. PMID- 8247173 TI - [No advantage in the addition of dipyridamole or of oral anticoagulants in comparison to low-dose acetylsalicylic acid (50 mg per day) in the prevention of venous transplant occlusion following coronary bypass surgery]. PMID- 8247174 TI - [Cervical myelopathy as complication of manual therapy in a patients with a narrow cervical canal]. PMID- 8247175 TI - [Acute heart infarct? Open the involved vessel]. PMID- 8247176 TI - Potassium transport in the cortical collecting duct. PMID- 8247177 TI - Influence of immunoadsorption on the removal of immunoglobulin G autoantibodies in crescentic glomerulonephritis. AB - We report the treatment of 5 patients with crescentic glomerulonephritis by immunoadsorption using a protein A column. Two had systemic vasculitis, 2 antiglomerular basement membrane disease and 1 systemic lupus erythematosus (SLE). In the patients with systemic vasculitis and SLE, there was successful removal of autoantibodies and rapid control of disease; remission was maintained over a mean follow-up of 2 years. Clinical improvement was not seen in 2 patients with anti-glomerular basement membrane (GBM) disease who presented with dialysis dependent renal failure. There were no apparent clinical side-effects related to the immunoadsorption procedure. Protein A immunoadsorption offers a semiselective alternative to plasma exchange and appears to be safe and efficient in removing pathogenic autoantibodies in crescentic glomerulonephritis without anti-GBM antibodies. PMID- 8247178 TI - Proteinuria selectivity index--prognostic value in lipoid nephrosis and related diseases. AB - In order to predict the steroid response in lipoid nephrosis (LN), we studied age, sex, proteinuria level, histological features and proteinuria selectivity index (SI; ratio between IgG and transferrin clearances) in 52 LN cases (minimal change disease: n = 39; focal glomerulosclerosis+IgM nephropathy: n = 13). The multivariate analysis showed that age, sex and proteinuria level were not contributive, whereas histology and SI were. The predictive value of SI was much higher than that of histological type (McFadden's r2: 47% vs. 22%, p < 0.001). Thus, SI should be systemically assessed in idiopathic nephrotic syndrome for reviewing the pathologic classification obtained by histology. However, if its prognostic value is lower than that of selectivity, initial renal biopsy remains necessary for diagnosis in adults. PMID- 8247179 TI - Study of asymptomatic microscopic hematuria in potential living related kidney donors. AB - Thirty potential living related kidney donors with asymptomatic microscopic hematuria of nonsurgical causes were entered in this study. They underwent thorough history taking, medical and ENT examination, laboratory and radiologic assessment and pure-tone audiometry. Family members were also subjected to urine analysis and audiometry. Moreover, the 30 donors were subjected to kidney biopsies which were examined by light microscopy, direct and indirect immunofluorescent microscopy, and electron microscopy. Hereditary nephritis (with or without sensorineural deafness) was found to be the most common cause of asymptomatic microscopic hematuria (25/30), followed by isolated C3 deposits disease (3/30), IgA nephropathy (1/30) and IgM nephropathy (1/30). Since these disease conditions are of a progressive nature, we have concluded that relatives of uremic patients with asymptomatic microscopic hematuria should not be considered for kidney donation even if they are strongly motivated. PMID- 8247180 TI - Experimental immunoglobulin A nephropathy induced by gram-negative bacteria. AB - A study was conducted to determine whether intraperitoneal and oral administration of formalin-fixed gram-negative bacteria induced immunohistologically and ultrastructurally evident glomerular deposition of IgA and C3 in C3H/HeN mice. Separate treatments with strains of Pseudomonas aeruginosa, Escherichia coli, Haemophilus influenzae, Klebsiella pneumoniae, and two kinds of lipopolysaccharide (LPS) were used. Two mice in each treatment group were sacrificed at 10, 20 and 30 weeks of age to examine sequential glomerular changes. In addition to the intraperitoneal administration (IP) groups receiving whole formalin-fixed bacterial cells, cell precipitate and supernatant fractions of each bacterial strain after sonication were injected intraperitoneally once a week, and the mice were sacrificed at 30 weeks of age. Sequential quantitation or IgG, IgA or IgM in serum and the isotypes specific for each of the bacterial strains or LPS administered was performed by ELISA. The incidence of immunofluorescence positivity for glomerular IgA and C3 was 37-71 and 37-66.7%, respectively, in the IP groups that had received bacterial cells of each strain, which was significantly higher than that in the IP groups given LPS or in the controls. These results suggest that cell wall components common among gram negative bacteria, other than LPS, play a major role in the glomerular deposition of IgA and C3. This is the first use of gram-negative bacteria to establish an active model of IgA nephropathy. PMID- 8247181 TI - Prevalence of glomerular hyperfiltration and nephromegaly in normo- and microalbuminuric type 2 diabetic patients. AB - Glomerular hyperfiltration, correlated with nephromegaly, is a frequent finding in type 1 (insulin-dependent) diabetes. In type 2 (non-insulin-dependent) diabetes, very few studies have been performed, and the results have been inconclusive. Glomerular filtration rate (GFR) and kidney volume, using 99mTc DTPA scintigraphy and ultrasonography, respectively, were evaluated in 58 control subjects and 163 type 2 diabetic patients; 79 of whom were normoalbuminuric and 84 microalbuminuric. In the two groups of patients, these parameters did not differ significantly from those of controls, even when hypertensive subjects were excluded. Glomerular hyperfiltration was observed in 10 cases; all were normotensive (9.8%), of whom 7 were normoalbuminuric and 3 microalbuminuric. Nephromegaly was observed in 3 other normotensive microalbuminuric diabetic patients. Hypertensive subjects showed a lower GFR than normotensive patients and control subjects. Multivariate analysis showed a negative correlation between glomerular filtrate and systolic blood pressure (BP) in the overall population of patients and in normo- and microalbuminuric patients taken separately. It is concluded that the relationship between these variables forms a continuum in our type 2 diabetic patients; it may also be important in determining the low prevalence of hyperfiltration and nephromegaly found in our patients, who had BP levels higher than those of controls. PMID- 8247182 TI - The reliability and validity of echocardiographic measurement of left ventricular mass index in hemodialysis patients. AB - We assessed the reliability and validity of a formula, based on echocardiographically derived parameters, to calculate left ventricular mass index (LVMI) in a group of 15 chronic hemodialysis patients. All patients had M mode echocardiography before and after a hemodialysis session. Echocardiograms were interpreted by 2 observers blind to each other's measurements. Interobserver reliability for LVMI was high (r = 0.94, p < 0.0007). LVMI decreased in 11 of 15 patients during dialysis and increased in 4. The mean difference in LVMI between pre- and posthemodialysis was 26.2 +/- 15 g/m2 (p < 0.0001). End-diastolic diameter decreased from 53.5 +/- 5.9 to 49.5 +/- 7.5 mm (p = 0.0016). These data indicate that measurement of LVMI is highly reproducible in hemodialysis patients but that it changes significantly over the course of a hemodialysis session. Its use as an outcome measure in clinical trials in hemodialysis patients should be interpreted with caution. PMID- 8247183 TI - Mononuclear cell subsets in IgM mesangial proliferative glomerulonephritis. A comparative study with minimal change nephrotic syndrome and immunonegative mesangial proliferative glomerulonephritis. AB - Glomerular and interstitial leukocyte subpopulations were analyzed in renal biopsies from 18 patients with IgM mesangial proliferative glomerulonephritis (IgM MPGN), 19 patients with minimal change nephrotic syndrome (MC) and 10 patients with immune-negative mesangial proliferative glomerulonephritis (IN MPGN), by immunoperoxidase techniques with monoclonal antibodies. Mesangial cell proliferation was strongly correlated with absolute numbers of intraglomerular T lymphocytes (r = 0.71; p < 0.01) in IgM MPGN, but not in MC or IN MPGN. Significant differences were found in the numbers of macrophages, CD4- and CD8 positive glomerular cells (Student's t test p < 0.01, 0.05 and 0.01, respectively) in IgM MPGN, but not in MC or IN MPGN. The numbers of CD45-, CD3- and CD8-positive cells also differed in each patient group (ANOVA p < 0.01, 0.05 and 0.05, respectively), the greatest and smallest values appearing in IgM MPGN and MC, respectively. Multiple regression test showed initial proteinuria values in IgM MPGN to be closely dependent on the density of neutrophils, macrophages, T and B lymphocytes and CD4 cell inflammatory infiltrates (r2 = 0.92; p < 0.01). At the end of the follow-up, proteinuria in IgM MPGN, but not in MC or IN MPGN, was dependent on T cell infiltrate (r2 = 0.97; p < 0.01). Our findings suggest that proteinuria in IgM MPGN results from local mesangial damage rather than from the effects of a soluble circulating factor, as has been proposed for MC. The clinical and immunohistochemical differences observed between these two processes support the notion that they should be considered as separate entities. PMID- 8247184 TI - 31P magnetic resonance spectroscopy investigation of skeletal muscle metabolism in uraemic patients. AB - Using the method of in vivo magnetic resonance spectroscopy we examined 17 patients with moderately advanced chronic renal insufficiency, 21 patients with chronic renal failure treated by haemodialysis, and 15 dialyzed patients with symptomatic renal osteopathy. The ratios of intracellular phosphocreatine and inorganic phosphate concentrations of these subjects measured at rest were compared with those found in healthy controls. While we noted significantly lower (p < 0.01) ratio values in all patients, subjects with osteopathy showed a lower value than dialyzed patients free of bone disease. Haemodialysis improved the result of examination in 7 patients. The results can be summarized as follows: (1) patients with altered renal function have significantly impaired energy metabolism of skeletal muscle, and (2) the disorder is more severe in patients with renal osteopathy than in those free of it. PMID- 8247185 TI - Tubular secretion of Tamm-Horsfall protein is decreased in type 1 (insulin dependent) diabetic patients with diabetic nephropathy. AB - Signs of glomerular, proximal and distal tubular dysfunction as well as metabolic control were studied in type 1 diabetes mellitus. To that end, the urinary excretion rates of albumin, sodium, phosphate and Tamm-Horsfall protein as well as HbA1c levels were measured in 20 patients with different degrees of diabetic nephropathy (positive Albustix for several years). Eight diabetic patients with short duration of diabetes and without any diabetic complications and 10 apparently healthy subjects were studied for comparison. The HbA1c levels in the three groups were 8.6 +/- 1.2, 5.9 +/- 2.2 and 4.1 +/- 0.4%, respectively (mean +/- SD). Duration of diabetes in the two diabetic groups were 27 +/- 7 and 3 +/- 1 years, respectively. The urinary protein levels were measured by enzyme-linked immunoassays. The fractional clearance of sodium (1.9 +/- 1.9%; p < 0.001) and phosphate (27 +/- 11%; p < 0.01) were increased in patients with diabetic nephropathy compared to diabetic patients without nephropathy (0.6 +/- 0.2 and 16 +/- 4%) and healthy control subjects (0.6 +/- 0.1 and 16 +/- 4%, respectively). Tamm-Horsfall protein excretion rate was decreased in both diabetic groups (15.0x/3.1 and 37.9x/1.9 micrograms/min, geometric mean x/tolerance factor, p < 0.001 and p < 0.05, respectively) compared to the healthy subjects (63.8x/1.3 micrograms/min). Furthermore, patients with diabetic nephropathy had a lower excretion rate of Tamm-Horsfall protein (15.0x/3.1 micrograms/min) compared to patients without signs of nephropathy (37.9x/1.9 micrograms/min, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247186 TI - Effect of cyclosporin A on renal cortical resistances measured by color Doppler flowmetry on renal grafts. AB - Doppler spectra were recorded at different cyclosporin A (CSA) levels (trough and peak) in 30 stable renal-transplanted outpatients: 15 with unimpaired renal function (plasma creatinine < 150 mumol/l) and 15 with renal impairment (plasma creatinine 150-350 mumol/l). Pulsatility (PI) and resistive indexes (RI) have been measured in the renal artery at the hilum and in the renal cortex. RI and PI were markedly increased (p < 0.0001) in the cortex at peak time while in the renal artery no significant changes were observed. These variations were statistically related with CSA blood levels (PI = p < 0.02; r = 0.54, RI = p < 0.05; r = 0.45). These effects were also found in the presence of renal damage. CSA dose-dependently reduces cortical blood flow, causing a persistent arteriolar vasoconstriction and a reduction in diastolic flow. This effect can be measured in man in a noninvasive and repeatable way using color Doppler sonograms. PMID- 8247187 TI - Anomalous rise of serum osteocalcin following desferrioxamine treatment in aluminium intoxication. AB - A patient with impaired renal function, severe osteomalacia and aluminium intoxication is described. In response to Desferal (desferrioxamine) treatment, serum osteocalcin rose 8-fold whereas serum alkaline phosphatase and procollagen I peptide levels changed little. Chromatographic separation showed that the measured osteocalcin coeluted with intact osteocalcin. The osteocalcin in the serum probably comes from the liberation of pre-existing osteocalcin from the bone, concomitant with aluminium mobilisation, rather than by stimulation of de novo synthesis. PMID- 8247188 TI - Effect of long-term hemodialysis on arterial compliance in end-stage renal failure. AB - To assess the influence of long-term hemodialysis on arterial compliance, the elastic vessel wall properties of the common carotid artery were determined in 20 normotensive renal transplant recipients (age 44.7 +/- 4.1 years) 8-12 weeks after first transplantation and in 10 healthy controls (age 45.9 +/- 5.2 years). Arterial distension was measured by using a multigate pulsed Doppler system, blood pressure curve was recorded by finger-plethysmography. 10 patients with a prior long-term hemodialysis of 51 +/- 11 months were compared to 10 patients with a prior short hemodialysis of 18 +/- 7 months. The patients and controls had been matched in respect of age, sex and blood pressure. In the long and short term hemodialysis group, the proportion of patients (n = 10) with a history of mild hypertension was similar--mild hypertension for 25 +/- 10 months (n = 5) and for 27 +/- 9 months (n = 5). In the group with long-term hemodialysis, the cross sectional compliance and the distensibility coefficient was significantly reduced in comparison to the group with short-term hemodialysis (p < 0.005) and to the control group (p < 0.001). A significant inverse correlation between the hemodialysis period and the distensibility coefficient (r = -0.59; p < 0.005) showed a decrease in arterial compliance with the length of hemodialysis treatment. The results demonstrate that vessel wall elasticity decreases with the length of hemodialysis treatment and is reduced by hemodialysis-dependent factors, which are detached from sustained arterial hypertension. As cause of reduced arterial compliance in long-term hemodialysis hypervolemia, hypercirculation and disturbed calcium-phosphate metabolism is suggested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247189 TI - Effect of prednisone on renal function in man. AB - To clarify the rise in plasma creatinine concentration previously observed during prednisone treatment, we studied changes in renal function in Graves' ophthalmopathy patients before and after 2 weeks of either prednisone 60 mg/day or retrobulbar radiotherapy (controls). Compared to retrobulbar radiotherapy, prednisone treatment was associated with an increase in: (a) plasma creatinine concentration (from 68 +/- 4 to 76 +/- 4 mumol/l), (b) glomerular filtration rate (GFR, from 93 +/- 4 to 102 +/- 5 ml/min/1.73 m2), and (c) urinary creatinine excretion rate (from 510 +/- 40 to 570 +/- 40 mumol/h). We conclude that GFR rises during 2 weeks of high-dose prednisone administration, a rise that is not reflected by a decrease in plasma creatine concentration. On the contrary, both plasma creatinine concentration and urinary creatinine excretion increase, probably as a result of the catabolic effect of prednisone. As established by the present study, prednisone 60 mg/day is associated with protein wasting, also after 14 days of treatment. PMID- 8247190 TI - Peritoneal dialysis fluid inhibition of polymorphonuclear leukocyte respiratory burst activation is related to the lowering of intracellular pH. AB - In order to elucidate the mechanism of peritoneal dialysis fluid inhibition of cell functions, laboratory-prepared fluids were used to investigate the specific influences of low pH and high lactate concentration on neutrophil viability, phagocytosis, respiratory burst activation and leukotriene B4 (LTB4) generation. In the absence of any reduction of viability, respiratory burst activation, stimulated by serum-treated zymosan (STZ), was significantly inhibited by fluids of low pH containing high concentrations of sodium lactate. Neither low pH nor lactate concentration alone, however, caused significant suppression of this parameter of cell activation. Under the same conditions, the phagocytosis of STZ was partially inhibited in a lactate- and pH-dependent manner. In contrast, the generation of LTB4 in response to STZ was unaffected by pH and lactate concentration. The incubation of polymorphonuclear leukocytes (PMN) in fluids containing 35 mM lactate at pH 5.2 resulted in an immediate and profound lowering in intracellular pH ([pH]i) which was not observed in lactate-containing fluids at neutral pH or at low pH in the absence of lactate. We postulate that the critical lowering of [pH]i in PMN, caused by the combination of high lactate concentration and low pH of the dialysis fluids, is responsible for the observed inhibition of respiratory burst activation. It is also possible that under these conditions, the lactate ion acts as a proton carrier across the cell membrane following the [H+] gradient. The time course of this [pH]i change suggests that host defence mechanisms may be impaired following short-time exposure to unused dialysis fluid prior to its equilibration in vivo. PMID- 8247191 TI - Response of vasoactive substances to intermittent ultrafiltration in normotensive hemodialysis patients. AB - The changes in blood volume (BV), atrial natriuretic peptide (ANP), plasma renin activity (PRA), aldosterone (Aldo), norepinephrine (NE), epinephrine (Epi), parathyroid hormone (PTH), arginine vasopressin (AVP) and the cyclic nucleotides cAMP and cGMP were measured during a fluctuating BV cycle in 15 patients with end stage renal failure maintained on chronic hemodialysis (HD). HD consisted of 4 periods of about 60 min each. The first half of each HD period consisted of ultrafiltration (UF) greater than 1,000 ml/h, and the second half consisted of no UF. Changes in relative BV were measured using continuous hemoglobinometry. Total BV at the end of treatment was 74.3 +/- 6.9% of the pretreatment volume. A significant positive correlation between BV and the levels of ANP, PTH, Epi and cGMP and an inverse correlation between BV and PRA, Aldo, AVP and NE were demonstrated. While mean values of NE and AVP levels were directly related to actual changes in BV, individual values did not homogeneously reflect this relationship. The cyclic nucleotides cGMP and cAMP did not follow immediate BV changes, but showed a significant decrease correlated with diminished BV. Based on a pre-postdialysis analysis, significant changes in PRA and Aldo were missing. It seems possible that vascular stability in dialysis patients may be maintained by the response of NE and AVP, and not by the renin-aldosterone system. The changes in ANP and cGMP values correlated most significantly (r = 0.38 and r = 0.51, p < 0.005) with the changes in BV, but no single variable could explain the blood pressure regulation during HD with intermittent rapid UF. PMID- 8247192 TI - Production of tumor necrosis factor alpha and hemodialysis. AB - The production of TNF alpha by peripheral blood mononuclear cells (PBMC) was determined in 18 hemodialysis (HD) patients. Blood was taken from each patient before and after an HD treatment. Both pre- and post-HD PBMC produced significantly more TNF alpha than controls (TNF alpha units/ml; mean +/- SEM; controls 3.1 +/- 0.7; pre-HD 9.7 +/- 3.9; post-HD 19.8 +/- 7.7, p < 0.05). In addition, post-HD PBMC produced significantly more TNF alpha than pre-HD PBMC suggesting that the HD procedure itself may activate cytokine production. This was true when PBMC were cultured in serum free medium as well as on culture with non-HD sera (human AB) and autologous sera. A positive correlation was also found between the production of TNF alpha and age in HD patients (r = 0.58; p < 0.01). Finally, normal PBMC cultured in post-HD sera produced significantly less TNF alpha than when cultured in the same sera pre-HD (p < 0.02). These findings suggest that PBMC of HD patients are chronically stimulated to produce TNF alpha which may contribute to some of the short-term and long-term complications of HD. PMID- 8247193 TI - Epidermal Langerhans cells in uremic patients on hemodialysis or continuous ambulatory peritoneal dialysis. AB - Skin biopsies of 33 uremic patients-13 patients on continuous ambulatory peritoneal dialysis (CAPD), 12 on hemodialysis (HD), 8 patients with end-stage renal disease (ESRD) before initiation of dialysis treatment-and 10 healthy volunteers were investigated to determine the number of Langerhans cells (LC) by light microscopy after staining for S-100 protein. The epidermal LC count was significantly lower in patients on CAPD (mean: 62.9 LC/mm2; p = 0.027) and patients on HD (mean: 30.4 LC/mm2; p = 0.0015) compared to controls (mean: 110.1 LC/mm2) and uremic patients before initiation of dialysis treatment (mean: 122.6 LC/mm2). The difference between LC counts of CAPD and HD patients did not reach statistical significance (p = 0.057). There was no relation between LC count and age (p = 0.057) or epidermal width (p = 0.26). No statistically significant correlation could be demonstrated between duration of dialysis and LC count (r = 0.33, p = 0.10). LC counts of CAPD patients with diabetes mellitus (n = 7) were not significantly different from those of nondiabetics (n = 6; p = 0.77). LC counts seem to be normal in uremic patients before dialysis treatment. The reduction in LC density in the skin of dialysis patients may contribute to immunodeficiency of uremic patients on regular dialysis treatment. PMID- 8247194 TI - Comparison of sublingual captopril, nifedipine and prazosin in hypertensive emergencies during hemodialysis. AB - Hypertensive emergencies in hemodialysis require immediate therapy, usually by parenteral drug administration; however, sublingual medications may have potential in this special condition. Sublingual captopril (25 mg), nifedipine (10 mg) and prazosin (2 mg) were prescribed to determine the effectiveness and safety of each medication in the treatment of hypertensive emergencies during hemodialysis. Blood pressure and heart rate were measured continuously up to 120 min postdose. The response rates were 83% for captopril, 90% for nifedipine and 11% for prazosin. The significant hypotensive effects of both sublingual captopril and nifedipine occurred at 10 min and continued up to 120 min. The reduction of systolic blood pressure occurred earlier in nifedipine than captopril (10 vs. 15 min). No significant difference in heart rate between them was noted. There were no side effects in the captopril group but flushing, tachycardia and headache were observed in 4 patients of the nifedipine group. We concluded that sublingual captopril and nifedipine were effective but captopril seemed to have less side effects than nifedipine and may be an excellent alternative to sublingual nifedipine in the urgent treatment of hypertensive emergencies in hemodialysis. Prazosin was not recommended because of its low response rate. PMID- 8247195 TI - Oxidative injury to erythrocytes, cell rigidity and splenic hemolysis in hemodialyzed patients before and during erythropoietin treatment. AB - The oxidative injury to erythrocytes, red blood cell (RBC) rigidity and splenic hemolysis was assayed in 17 chronically hemodialyzed patients before and during recombinant erythropoietin (EPO) treatment. When a stable hematocrit between 30 and 35% had been established for at least 4 months, a statistically significant increase in RBC volume, hemoglobin concentration, hematocrit, reticulocyte count, and several RBC enzymes (2,3-diphosphoglycerate, glucose 6-phosphate dehydrogenase, pyruvate kinase, hexokinase) was noted. This indicated significant RBC rejuvenation under the influence of EPO. However, no significant improvement in the RBC oxidative sensitivity, RBC deformability, splenic RBC volume, slow mixing splenic RBC volume, and the intrasplenic RBC transit time could be disclosed. These data confirm the existence of an extra-erythrocytic factor in uremic plasma, which is partly responsible for a reduced RBC life span in hemodialysis patients despite EPO treatment. PMID- 8247196 TI - Clinical evaluation of 99mTc diaminocyclohexane, a renal tubular agent with cationic transport: results in healthy human volunteers. AB - As alternatives to anionically transported hippuran, structure distribution experiments on a series of 99mTc-labelled primary substituted ethylene diamine compounds led to selection of 99mTc diaminocyclohexane (DACH) for clinical evaluation, 99mTc DACH, a cation with the structure trans-[O2(DACH)2 99mTc]+ is prepared by mixing 50 mumol DACH, 1 mumol stannous tartrate and 500 MBq of 99mTc. Seven normal volunteers underwent renal imaging and clearance studies using 150 MBq of 99mTc DACH and 1 MBq of 125I hippuran simultaneously. The images with 99mTc DACH revealed good uptake and excretion. The mean +/- 2 SD values of parenchymal and whole-kidney transit time indices and mean parenchymal transit time were 46 +/- 33, 75 +/- 64 and 141 +/- 51 s, respectively, similar to mercaptoacetyl triglycine. The mean clearance of 99mTc DACH was found to be 163 ml/min (SD = 32). Following cationic blockade with 900 mg oral thiamine, significant reduction (p < 0.001) in DACH clearance was noted, but hippuran clearance remained unaltered. The results support the hypothesis that 99mTc DACH is transported cationically. PMID- 8247197 TI - Blood and graft eosinophilia as a rejection index in kidney transplant. AB - The relevance of eosinophilia in the physiopathology of transplant rejection has yet to be established. The appearance of eosinophilia has been occasionally associated with an adverse prognosis on graft rejection episodes. The aim of the present study was to evaluate the role and prognostic implications of blood and graft eosinophilia in kidney transplant rejection. We have examined the intrarenal infiltrate in 173 fine-needle aspiration biopsies from 36 consecutively transplant patients, and blood samples obtained simultaneously with fine-needle aspirations. Two different immunosuppressive regimens were administered: triple therapy (azathioprine + prednisone + antilymphocytic globulin) in patients with posttransplant acute tubular necrosis and cyclosporine A monotherapy in the rest of the patients. Comparing the two immunosuppressive groups, more elevated eosinophil values were observed in the monotherapy group during stable graft and also at the rejection episode. In the monotherapy group, a significant increase in the eosinophil values, in peripheral blood samples and in the intragraft infiltrates were noted at the rejection episode with respect to the stable situation. Following pulsed-steroid treatment an immediate disappearance of the eosinophils was evident. In contrast, no differences could be demonstrated between these two clinical situations in the TT group. Higher rates of eosinophils in the intrarenal infiltrate with respect to peripheral blood samples were observed during rejection episodes, suggesting some role of the eosinophils in the physiopathology of graft rejection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247198 TI - Simultaneous occurrence of IgA nephropathy and sarcoidosis in the context of pre existent minimal change nephrotic syndrome. AB - Simultaneous occurrence of IgA nephropathy and sarcoidosis is described in a 42 year-old Asian female, who earlier had a steroid-resistant, no light microscopy change nephrotic syndrome that resolved spontaneously. Occurrence of three, apparently unrelated conditions in this case may be fortuitous, or causally related. We discuss the possible causal relationship amongst these conditions. PMID- 8247199 TI - Focal segmental glomerulosclerosis in a patient with a family history of the disease. PMID- 8247200 TI - Membranous glomerulonephritis associated with autosomal dominant polycystic kidney disease. PMID- 8247201 TI - Visceral leishmaniasis-like cause of unexplained fever in a reno-pancreatic graft recipient. PMID- 8247202 TI - Antineutrophil cytoplasm and antielastase antibodies in hemolytic-uremic syndrome. PMID- 8247203 TI - Renal amyloidosis in a patient with epidermal inclusion cysts. PMID- 8247204 TI - Calcium alginate versus aluminum hydroxide in patients on hemodialysis. PMID- 8247205 TI - Recurrence of thyrotoxicosis after correction of uraemia in a patient with chronic renal failure. PMID- 8247206 TI - Pulmonary tumor embolization after renal transplantation. PMID- 8247207 TI - Angiotensin I-converting enzyme levels and renin-aldosterone axis recovery after cessation of chronic licorice ingestion. PMID- 8247208 TI - Successful birth in a hemodialysis patient with SLE. PMID- 8247209 TI - Interaction between cyclosporin and chloroquine. PMID- 8247210 TI - More about tumoral calcinosis in maintenance hemodialysis patients. PMID- 8247211 TI - [Chemonucleolysis and lumbar disk herniation]. PMID- 8247212 TI - [Percutaneous treatment of lumber disk herniation. Radiological criteria of therapeutic decision]. AB - Invasive diagnostic imaging methods such as discography, discomanometry, C.T. discography are generally not required to accurately depict herniated lumbar disks. On the contrary they should be implemented in specific clinical situations in order to confirm the choice of a percutaneous treatment: nucleolysis or nucleotomy. The major role is devoted to discography. Discomanometry and C.T. discography are second order methods that will help in difficult cases in choosing therapeutic strategy. PMID- 8247213 TI - [Foraminal and latero-foraminal hernia. Mid-term results of percutaneous techniques nucleolysis-nucleotomy]. AB - Forty cases of foraminal (28) and lateroforaminal (12) herniated discs treated either by nucleolysis (N.L.) or percutaneous automated nucleotomy (P.A.N.) were investigated over a period of 12 months or more. The group of patients treated with P.A.N. (15 males, 5 females) with a mean age of 41.8 years, had severe radicular pain in 25% of cases, caused by foraminal (15 cases) and lateroforaminal (5 cases) herniated discs which were predominantly located at the L4-L5 level (12 cases). The group of patients treated with N.L. (13 M, 7 F) with a mean age of 50.3 years complained of severe radicular pain in 65% of cases, caused by foraminal (13 cases) and lateroforaminal (7 cases) disc herniations evenly distributed over the last three mobile segments. The overall results obtained with N.L and P.A.N. showed a 67.5% success rate. The best results were obtained in cases of cruralgia with severe radicular pain caused by foraminal herniated disc at the L3-L4 or L4-L5 levels. A significantly better result was obtained in the group of patients treated by N.L. (80% success rate) than in the group of patients treated by P.A.N. (p < 0.04). PMID- 8247214 TI - [Percutaneous nucleotomy-diskectomy techniques. Automated and manual techniques. Indications and results]. AB - The authors review the percutaneous techniques: automated (Nucleotome) manual, with and without motorization ("shavers") with and without discoscopy. These techniques are indicated in less than 15% of cases of disc herniations; but, if the proper indications are respected, a rate of success between 75 and 85% can be expected. The guidelines of the indications are the following: contained disc herniation (longitudinal ligament not ruptured), subacute clinical course, no lateral spinal stenosis, age under 45.... Although the percutaneous techniques are simple and safe, they must be always preceded by a medical treatment during at least 6 weeks. The pre-operative imaging is of paramount value to assess the wholeness of posterior longitudinal ligament: computerized discography (C.T. scan+discography), M.R.I., disco-manometry.... The authors advocate for percutaneous discectomy versus chemonucleolysis, but surgical approach by micro technique remains indicated in most cases of lumbar herniated discs, especially in cases of ruptured discs (extruded or uncontained herniations), acute clinical course, spinal canal stenosis, age more than 45, or particular anatomical situations (such as deep-seated lumbosacral space or post-operative fibrosis). PMID- 8247215 TI - [Pituitary abscess]. AB - A pituitary abscess case clinically evolving as an adenoma is presented. The diagnosis was established at the time the surgical procedure was performed. Previous studies and postoperative investigation proved the absence of features accounting for side infection. Moreover, anatomopathological studies also dismissed the existence of any type of tumoral traces. However, due to a lack of experience, the pre-operative magnetic resonance imaging was interpreted as pertaining to pituitary adenoma on the basis of its greater frequency. PMID- 8247216 TI - [Sphenocavernous localization of meningeal neurosarcoidosis. Apropos of a case and review of the literature]. AB - The authors report on the case of a 32 year old woman treated with prednisone and diphenylsulfon for a multi-visceral sarcoidosis; headache and progressive oculomotor paralysis revealed an expansive lesion of the left cavernous sinus extending to the temporal fossa. C.T. scan, M.R.I., and angiographic data as well as the intra-operative findings evoked the diagnosis of meningioma. Pathological findings showed an epithelioid and gigantocellular granuloma without caseous necrosis, corresponding to a sarcoidosic lesion. After the operation, the increase of steroid doses was followed by a complete regression of the tumoral remnants, and total clinical recovery. In a patient having a previously diagnosed sarcoidosis with evidence of an intracranial tumor mimicking a meningioma, steroids should be first prescribed. Efficacy of steroids on sarcoid granulomas is often dramatic; surgical approach would be discussed in case of ineffective steroid therapy, of persisting questionnable diagnosis, and of tumoral threatening compression. PMID- 8247218 TI - [INSERM and research in neurosurgery]. PMID- 8247217 TI - [Perspectives of research in neurosurgery]. PMID- 8247219 TI - [Radionecrosis of the optic chiasma. A clinical and radiological study of 3 cases]. AB - The paper reports three cases of optic chiasmal radionecrosis due to radiotherapy of the sellar region. These three cases are compared with 34 ones found in literature. The use of M.R.I. is shown to dramatically improve diagnosis. Corticoids are useful in the treatment. The gravity of the complication, which implies visual prognosis, focuses on a strict use of radiotherapy. For benign or slowly evoluting lesions, surgery and/or regular controls by modern imaging techniques would often be more eligible than systematic earliest irradiation. PMID- 8247220 TI - [Giant ependymoma of the cauda equina. Long-term development apropos of 7 cases]. AB - Seven cases of giant ependymomas of cauda equina, extending from the conus medullaris to the sacrum, are reported. Mean age of the patients was 36 years. The initial symptoms were not specific, including low back pain and radiculalgias. However, in two cases, the clinical presentation was related to hydrocephalus. At the time of the operation, 6 patients presented with pain, weakness and sphincter dysfunction. The diagnosis of tumor was made after myelogram in all cases. Metrizamide enhanced CT scan has been performed in one case. None patient had preoperative magnetic resonance imaging (MRI) but MRI allowed the follow up in 5 patients and detection of local and remote recurrences in 4 cases. All the tumors were attached to the filum terminale, and unless extended from L2 to S1. Pathologically, 3 patients had tumors classified as myxopapillary and 4 of the cellular type. Two patients had gross total resection of the tumor at the initial operation. Five patients had initial biopsy to make a diagnosis and required subsequent surgery for radical excision after an average of 16.8 months. Radiation therapy has been performed in 1 case after biopsy, in 1 case after total resection (grade III) and in 2 cases after recurrences. Three patients died 1 year, 5 years, 12 years post-operatively. Among these 3 patients, 2 had recurrences. Four patients are alive. One patient has no recurrence 5 years after initial surgery. Another patient remains symptom free 7 years after surgery but MRI showed a local recurrence. The last two patients present recurrences 7 years and 8 years after surgery. These recurrences have been treated by surgical removal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247221 TI - [Surgical applications of anatomical variations of the median nerve at the wrist]. AB - After a brief report of median nerve anatomy at wrist, authors describe terminal variations of its distribution. Classic description of five terminal sensitive and motor median nerve branches is not constant. Many anatomic variations have been reported and so many, classifications proposed. Lantz' classification is useful, it may be divided in four groups. Other anatomic variations concern cutaneous palmar nerve branch and different nervous anastomosis. Advantage of this anatomic study is incidence of variations on surgical access of carpal tunnel. Those nervous variations of median nerve at wrist are frequent, unknown and wrong indexed and may be so many "anatomic traps" for surgeon. PMID- 8247222 TI - B-50/GAP-43 phosphorylation in hippocampal slices from aged rats: effects of phosphatidylserine administration. AB - Phosphorylation of the presynaptic protein B-50/GAP-43, a substrate of protein kinase C (PKC), has been implicated in neuronal mechanisms related to learning and memory. We evaluated both basal (5 mM KCl) and stimulated (30 mM KCl) B 50/GAP-43 phosphorylation in 32P-prelabeled hippocampal slices obtained from adult and senescent male Sprague-Dawley rats. The in situ B-50/GAP-43 phosphorylation was assayed by quantitative immunoprecipitation. There was no age related difference in B-50/GAP-43 basal phosphorylation. However, B-50/GAP-43 phosphorylation in depolarized slices from aged rats was significantly decreased relative to that of adult animals. Aged rats were treated with either tris buffer or sonicated suspension of phosphatidylserine (PS) in tris buffer (15 mg/kg IP for 7 and 17 days). PS did not affect basal and high K(+)-induced B-50/GAP-43 phosphorylation in the 7-day treatment. However, after 17 days, PS restored the K(+)-induced B-50/GAP-43 phosphorylation. It is proposed that repeated PS administrations might be beneficial to the age-induced deterioration of endogenous B-50/GAP-43 phosphorylation by acting on Ca++ homeostatic mechanisms and/or PKC. PMID- 8247223 TI - Clinical comparison of Alzheimer's disease in pedigrees with the codon 717 Val- >Ile mutation in the amyloid precursor protein gene. AB - Alzheimer's disease (AD) is the most common cause of dementia (32). Although the majority of cases of AD are sporadic, the most consistent risk factor detected in several epidemiological studies has been a positive family history of the disease (14,21). In addition, many large pedigrees have been described in which AD appears to be inherited as an autosomal dominant disorder. In one such pedigree (F23) a point mutation within the beta-amyloid precursor protein (APP) gene at codon 717 was identified and hypothesized to be pathogenic (10). The mutation results in a valine to isoleucine change in APP (APP717 Val-->Ile). Subsequent screening has revealed four other pedigrees, detailed in this study, in which this mutation co-segregates with AD (13,26,37). In addition, one other pedigree (Tor3) with this mutation has been described (15) and detailed clinical, neuropsychological, and neuropathological data are reported. Tor3 is discussed below in comparison to the findings in the families in this study. The five families we report with the mutation were identified in Britain (1 family), the United States (1 family), and Japan (3 families). The mutation has not been reported in the general population of any of these countries (3,13,26,33). On this basis alone it seems this mutation is pathogenic. Other APP codon 717 mutations have been identified which co-segregate with the disease (4,25). Also, a double mutation in APP at codons 670/671 has been shown to cosegregate with the disease in two large Swedish pedigrees (22). In all cases, there is complete co segregation of the APP mutation with early onset AD, providing overwhelming statistical evidence that these mutations are pathogenic. We present the clinical features and limited neuropathology of AD in these families with the APP 717 Val- >Ile mutation. PMID- 8247224 TI - GFAP mRNA increases with age in rat and human brain. AB - Glial fibrillary acidic protein (GFAP) mRNA was examined by RNA blot hybridization in three age groups of two cohorts of male F-344 rats and in 47 human postmortem brain samples. GFAP mRNA increased in the hippocampus and striatum of 24 versus 6- to 7-month-old rats. Another astrocytic molecular marker, glutamine synthetase mRNA, did not change with age in rat brain. Rat GFAP mRNA prevalence was inversely correlated with serum testosterone but not correlated with serum corticosterone. In human hippocampus, frontal and temporal cortex, GFAP mRNA also increased in older (60-79 years) compared with middle-aged (25-59 years) individuals. In contrast, mitochondrial cytochrome oxidase subunit 1 mRNA did not change between age groups in any region. By combining the three regions for further analysis, GFAP mRNA increased with age irregardless of gender, alcoholism in the middle-aged group, or whether brains were classified as normal or neuropathologic (excluding Alzheimer's disease pathology). These data indicate that increased GFAP protein or GFAP-immunoreactive astrocytes in rats and humans may result from transcriptional or post-transcriptional regulation and extend the number to three species (including mouse) showing an increase in GFAP mRNA with age. Factors that are known to regulate GFAP mRNA expression in young brains are considered as possible causes of age-related increases. PMID- 8247225 TI - Chronic treatment of aged mice with L-deprenyl produces marked striatal MAO-B inhibition but no beneficial effects on survival, motor performance, or nigral lipofuscin accumulation. AB - Male C57BL/6J mice were provided I-deprenyl (at 0, 0.5 mg/kg or 1.0 mg/kg per day) in their drinking water beginning at 18 months of age. A battery of motor tests, including open-field, tightrope, rotorod, inclined screen, runwheel, and rotodrum tests, was administered before treatment and then 6 months later at 24 months of age. A subsample of mice was retested again at 27 months of age. An untreated group of 9-month-old mice served as young controls. Deprenyl treatment reduced striatal MAO-B activity by up to 60% after 6 months on treatment but had no significant effects on striatal catecholamine levels. No significant effects of deprenyl treatment were observed on body weight, fluid intake, or survival of the mice. Chronic deprenyl treatment also did not affect motor performance in any test, except rotodrum performance at 27 months of age, which was significantly better in the 1.0 mg/kg group treated group compared to controls. No age or deprenyl effects were observed with respect to cell counts in the substantia nigra. However, nigral cells containing lipofuscin increased with age, but this neurohistochemical parameter was also unaffected by deprenyl treatment. PMID- 8247226 TI - Light-induced gene expression in the suprachiasmatic nucleus of young and aging rats. AB - We investigated age-related changes in a molecular mechanism associated with synchronization of circadian rhythms to the environment. In mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus contains a circadian pacemaker that regulates a variety of physiological and behavioral rhythms. Recent studies have demonstrated photic induction of immediate early genes (IEGs) such as c-fos in the SCN in a circadian-phase dependent manner, suggesting that IEGs may be part of the pathway for entrainment of circadian rhythms. We find that there is a decreased response of the IEGs c-fos, and NGFI-A but not NGFI-B in the SCN of old animals after photic stimulation. Changes in gene transcription indicated by IEGs may provide insights into the molecular machinery of the biological clock and ultimately elucidate mechanisms underlying the age-dependent decay of circadian organization. PMID- 8247227 TI - Serum- and bradykinin-induced calcium transients in familial Alzheimer's fibroblasts. AB - The calcium-sensitive photoprotein, aequorin, was used to examine serum- and bradykinin-induced transient increases in free cytosolic calcium ions in skin fibroblasts from 10 individuals with early onset familial AD (FAD), including four who were biopsied before their clinical symptoms would allow a diagnosis of AD, 2 individuals with late onset FAD, 8 at-risk but nonsymptomatic individuals, and 13 controls. The data show that (a) among controls, the peaks of the calcium transients increase in height as a function of donor age; (b) transients induced by 10% serum, 10 nM bradykinin (BK) or 100 nM BK were generally lower in FAD fibroblasts, including those from donors in the early stages of the disease, than in age-matched control cells; (c) such transients are reduced in cells from a proportion of the nonsymptomatic, at-risk individuals. Thus, serum- and BK induced calcium transients are reduced in fibroblasts from both early and more advanced stage FAD donors and perhaps even from donors who are presymptomatic carriers of the defective gene. The data also suggest that changes in calcium transients in FAD fibroblasts neither mimic nor exaggerate the effects of normal aging. PMID- 8247228 TI - Morphologic alterations of choline acetyltransferase-positive neurons in the basal forebrain of aged behaviorally characterized Fisher 344 rats. AB - We examined Fisher 344 female rats aged 6, 27, and 33 months old. Prior to sacrifice and morphometric analyses of forebrain cholinergic neurons all rats underwent behavioral characterization in a spatial learning task using the Morris water maze. Performance on the spatial task permitted subsequent grouping of the 27- and 33-month-old animals into impaired or nonimpaired groups. Importantly, the percentage of animals that displayed spatial impairments increased sharply with advancing age. Quantitative assessment of the size and density of choline acetyltransferase (ChAT)-positive neurons throughout the basal forebrain revealed a significant enlargement of forebrain cholinergic neurons within 27-month-old nonimpaired rats compared to 6-month-old rats and 27- and 33-month-old impaired animals. This increase in size was most noted in the medial septum and nucleus of the diagonal band. Significant decreases in the density of ChAT-positive neurons was observed only in the nucleus of the diagonal band of 27-month-old impaired rats compared to 6-month-old controls. Although the significance of enlarged forebrain cholinergic neurons is unclear, we discuss the possibility that within aged rodents neuronal swelling is an active event and represents an early manifestation of the aging process and may constitute a restorative and/or compensatory event in that these rats are relatively asymptomatic with respect to their behavioral deficits. In addition, we discuss in some detail various technical and life effect issues which may vary the outcome of investigations of aged rodents. PMID- 8247229 TI - Incomplete achromatopsia in Alzheimer's disease. AB - We report that patients with Alzheimer's disease (AD) have a selective deficit in blue hue discrimination, as assessed with three clinical measures of color vision. The Farnsworth D-15 Test, the Lanthony New Color Test, and the City University Color Vision Test were administered to 32 patients with AD (ranging in dementia severity from mild to severe) and 32 age-matched normal control subjects (NCS). Of the AD patients, 11 who were representative of the larger group for age, education level, and dementia severity received a complete neuro ophthalmological examination that ruled out obvious disorders of the anterior visual structures. AD patients made significantly more tritan (blue) errors than NCS on all three color vision tests but did not make more protan (red) or deutan (green) errors on two of the three tests. The results support the conclusion that there is a deficit in color discrimination in AD that is specific to blue hues, and oppose the hypothesis that AD does not deleteriously affect the color opponent visual channel. In the absence of obvious damage to anterior visual structures, the likely substrates for the observed deficit are peristriate and inferotemporal visual cortices, which are subject to significant neuropathology in AD. PMID- 8247230 TI - Age-related changes in calcium homeostatic mechanisms in synaptosomes in relation with working memory deficiency. AB - Aging is associated with alterations in different systems that govern neuronal calcium homeostasis. This study was designed to determine whether any of these alterations may contribute to the decline in spatial working memory that is observed in old rats. Several parameters [initial (5 s) and steady state (15 min) 45Ca2+ uptake, FCCP-releaseable 45Ca2+, [Ca2+]i levels, depolarization-induced phosphoprotein (P97, PP65, P42) dephosphorylation and acetylcholine levels and release) involved in calcium homeostasis/signaling were determined in whole brain synaptosomes derived from adult (9-month-old) and old (24-month-old) rats that were evaluated for spatial memory performance in the eight-arm radial maze. The neurochemical analysis indicated that both the 9- and 24-month-old rats were impaired with respect to 3-month-old animals. When learners (animals reaching criterion; RC) were compared to memory impaired rats (MI), it was found that the FCCP-releaseable 45Ca2+ of synaptosomes, that reflects mitochondrial calcium, was lower in the MI than the RC rats and was correlated with the behavioral performance of the rats in their first testing sessions. The results suggest that the loss of calcium uptake capacity in synaptic mitochondria during aging may be associated with impaired working memory in old animals. PMID- 8247231 TI - Anticholinergic sensitivity in the aging rat septohippocampal system as assessed in a spatial memory task. AB - The effects of central cholinergic blockade on spatial memory were tested in aged and basal forebrain-lesioned rats using the Morris Water Maze. In Experiment 1, aged rats (18-21 months old) were characterized as behaviorally impaired or nonimpaired based on water maze performance prior to an atropine sulfate challenge. In the atropine test (50 mg/kg, IP), both the impaired and the nonimpaired rats showed a severe disruption of their search behavior compared to young subjects. This effect was due to blockade of central receptors since peripheral cholinergic blockade using atropine methylbromide did not produce any impairments. Experiment 2 investigated effects of atropine on rats with septal lesions (SL), nucleus basalis lesions (NBL), and rats with both lesions combined (SL + NBL). Before drug treatment, the groups with septal lesions (SL and SL + NBL groups) displayed a moderate impairment in locating the platform site. However, similar to the aged rats, the septal-lesioned rats exhibited severe impairments in the water maze during atropine treatment. This effect was not seen in the normal controls or in the NBL rats. Aged rats, either impaired or nonimpaired in a spatial memory task, showed a pronounced sensitivity to pharmacological blockade of central cholinergic neurotransmission which resulted in severe deficits in spatial navigation in the water maze. Since the same behavioral deficit was produced by cholinergic blockade in young rats with septal lesions, we concluded that the impaired water maze performance seen in the aged rats during cholinergic blockade resulted from impaired function in the septohippocampal system. PMID- 8247232 TI - Early age-dependent changes in noradrenaline efflux in the bed nucleus of stria terminalis: voltammetric data in rat brain slices. AB - Fast cyclic voltammetry (FCV) was applied to the detection of stimulated noradrenaline (NA) efflux in the bed nucleus of stria terminalis (BSTV) of the superfused rat brain slice from rats of different ages. Three age groups were compared: young (5 weeks; 130 +/- 8 g), adult (12 weeks; 308 +/- 12 g), and mature rats (36 weeks; 575 +/- 21 g). The effect of train duration (20-99 pulses at 100 Hz) and frequency of stimulation (50 pulses at 10-500 Hz) on NA efflux were examined. The effect of the alpha 2 antagonist yohimbine (1 microM) was also investigated. NA efflux was significantly (p < 0.05) greater in young than adult or mature rats on all train durations and stimulus frequencies tested: Maximal NA efflux on the longest train (99 pulses, 100 Hz, 10 mA, 0.2 ms) was 423 +/- 48 nM (young), 135 +/- 24 nM (adult), and 155 +/- 26 nM (mature). There were no significant differences between adult and mature rats. Yohimbine (1 microM) elevated NA efflux to a greater extent at lower (10 & 20 Hz) than higher frequencies in all age groups. Yohimbine also potentiated NA efflux more (p < 0.05) in young and adult rats than in mature animals. There were no significant differences between young and adult rats. The results indicate early maturational differences in both NA efflux and its control by alpha 2 adrenoceptors. Interestingly, the decreases in NA efflux and in yohimbine response were not temporally linked. PMID- 8247233 TI - Age-related decline in cholinergic synaptic transmission in hippocampus. AB - Age-dependent changes in central nervous system (CNS) cholinergic synaptic transmission were studied in three age groups of Sprague-Dawley and Fischer 344 rats: 1- to 2-month-old, 8- to 10-month-old, and 18- to 23-month-old. Utilizing intracellular recording techniques and the in vitro hippocampal slice preparation, we report an age-related decline in central cholinergic transmission as a function of age. Slow excitatory postsynaptic potentials (slow EPSPs) were reduced approximately 60% in aged (18- to 23-month-old) compared to younger (1- to 2-month-old) animals. The response of the postsynaptic membrane to the muscarinic agonist, carbachol (0.3 microM), was also reduced with age. These changes were not accompanied by a global decline in muscarinic receptor function since two additional measures of cholinergic function were not changed with age. Both presynaptic inhibition of fast excitatory synaptic transmission and postsynaptic inhibition of the afterhyperpolarization (AHP) following a train of spikes were not changed during aging. Our results suggest that a primary functional decline in central cholinergic mechanisms during aging may be a specific reduction in central cholinergic synaptic transmission. PMID- 8247234 TI - [Deletions within the gene of dystrophin in Duchenne and Becker muscular dystrophy]. AB - DNA was isolated and analysed in 96 patients with Duchenne or Becker muscular dystrophy (DMD, BMD); 9 of them were affected with BMD. Delections were found in 60 Patients (62.5%) using six cDNA probes. In some cases the PCR technique was also applied. In patients with BMD all deletions but one were in frame and involved exons 45-54. On the contrary, most deletions in DMD were out of frame and varied in their location. In five families prenatal diagnosis was carried out. PMID- 8247235 TI - [The assessment of amplitude and occurrence of F waves in hemiparetic patients]. AB - An analysis of occurrence of F response in a group of 40 patients with vascular unilateral hemiplegia was conducted. In trains of 100 supramaximal stimuli, the frequency and amplitude of the F wave was estimated bilaterally in the median and peroneal nerves. Differences in the appearance of those responses in groups with different muscular tone, were shown. Small changes were also observed contralaterally to the hemiparetic side. In 2 cases instead of F wave H reflex was recorded from the abductor pollicic brevis muscle. PMID- 8247236 TI - [Electronystagmographic examination in patients with amyotrophic lateral sclerosis (ALS)]. AB - 18 patients suffering from ALS underwent ENG examinations. Motor function of eyeballs in respect of optic and vestibular stimulus was evaluated. The study demonstrated that 8 patients exhibited lesion of central part of the vestibular system and in 2 cases features of peripheral type of vestibular disturbances were found. PMID- 8247237 TI - [Anticardiolipin antibodies in patients with cerebral ischemia]. AB - Six patients with high level of anticardiolipin antibodies were hospitalized for ischaemic strokes or transient ischaemic attacks. The clinical findings in these cases varied greatly. In three ischaemic episodes were recurrent. In two stroke developed at old age without prodromal symptoms. In two females the vascular involvement was generalized. Although the role of anticardiolipin antibodies in the pathogenesis of vascular changes of the central nervous system is not yet clear it is thought that they may be a risk factor for stroke. PMID- 8247238 TI - [Clinical assessment of therapeutic results of Ergolactyn (bromocriptine) in Parkinson's disease]. AB - The effect of Ergolaktyna (bromocriptine) produced by Grodzisk Pharmaceutical Works POLFA on the intensity of parkinsonism manifestations was studied in 27 patients in four clinical centres. All patients had moderately advanced parkinsonism and prior to Ergolaktyna they received L-Dopa preparations which were also given concomitantly with that preparation. The final assessment was done after 4 months of treatment in 18 cases. A good effect of the drug was found in 55% of cases (reduction of the intensity of parkinsonian signs, increased fitness and life activity). Adverse effects developed in 4 cases and required drug withdrawal. PMID- 8247239 TI - [Assessment of dexamethasone effectiveness in the treatment of ischalgia]. AB - The authors studied 43 patients admitted to hospital with acute episodes of ischalgia or recurrences caused by intervertebral disc disease. The patients were randomly selected into two groups treated with dexamethasone in high but rapidly decreasing doses, or tramadol. A comparison of the results in both groups showed that dexamethasone was effective and safe when given by this method to patients with ischalgia. PMID- 8247240 TI - [Benign spinal neoplasms: results of treatment and clinical aspects]. AB - Benign spinal neoplasm (b.s.n.) represented 11% (8 cases) of all histopathologically verified spinal neoplasm in 85 patients operated on between 1975 and 1990. Local back or neck pain were first clinical symptoms of b.s.n. On admission all the patients presented with bone lesions on X-ray plain radiograms of the spine. Surgical management included removal of the tumour and stabilization using bone grafts. Excellent results (complete pain relief and improvement of motor function making the patients ambulatory and independent) were obtained in all the cases. All the patients presented with solid fusion of bone grafts and good spinal stability at the last follow-up. PMID- 8247241 TI - [Aneurysmal bone cyst of the skull and vertebral column treated operatively]. AB - Aneurysmal bone cysts considered as a benign lesion are extremely rare in the skull or in the vertebral column. In the last 29-years were admitted to Department of Neurosurgery in Poznan 11 patients with aneurysmal bone cysts involving the skull in 4 and the spine in 7 cases. All but one patients were female, aged between 4 and 37 years. Location of skull cysts were: parietal in 1, occipital in 1, and parietotemporal in two cases. In patients with spinal affection the position of vertebral cysts were: cervical -2, dorsal -4 and lumbar -1 case. All patients suffered severe pain radiating to the adjacent area and showed local deformity of bone structure. Paraparesis or paraplegia were observed in cases with spinal involvement. In all patients, total surgical removal of the aneurysmal bone cysts was possible. Skull defect was covered with plastic materials. In the repair of the vertebral defect, pelvic bone grafts were used and fixed with wire suture. In follow-up ranging from 1 to 25 years there was no tendency for recurrence. PMID- 8247242 TI - [Cerebrospinal fluid rhinorrhea as a postoperative complication of pituitary tumor surgery. Corrective surgery of sella turcica by own methods]. AB - A relatively simple technically and effective own method of corrective operation of sella floor in cases of cerebrospinal fluid rhinorrhoea after transsphenoid operations for pituitary tumours is presented. This complication developed in 12 cases in a group of 720 patients operated on in the years 1973-1990. The described method was applied in 9 cases. PMID- 8247243 TI - [Regeneration of peripheral nerves by muscle autografts in rats. I. Morphological and morphometrical evaluation]. AB - Regeneration of myelinated axons with 1 cm freeze-thawed skeletal muscle autografts was compared with nerve grafts in the rats. Regeneration was assessed histologically and morphometrically in the grafts and distal nerve stumps. Recovery of axon numbers and fibre diameters in distal nerve stumps was similar in the two kinds of grafts 6 months after operation. These observations suggest that freeze-thawed autogenous muscle grafts might be used in the repair of peripheral nerves. PMID- 8247244 TI - [Neurotransplantation Part II. Autografts of adrenal medulla in Parkinson's disease]. AB - The paper reports the results of adrenal medulla autotransplantations into the brain of parkinsonian patients performed up to now in many neurosurgical centers in the world. Different surgical techniques and methods of tissue preparation for transplantation are also reviewed. Core Assessment Program for Intracerebral Transplantation (CAPIT) is presented. This program was developed as an unified system for patients' diagnosis and evaluation of clinical signs and symptoms before and after transplantation. It enables direct comparison of the results obtained in different centers. Surgical risk depending on the method of operation is discussed. The mechanisms involved in functioning of adrenal medulla grafts are considered as well as the trials to improve graft survival and its activity in the host brain. The results of neuropathological studies from autopsy cases are also reported. The analysis of about 400 published cases shows moderate or marked clinical improvement due to adrenal medulla autotransplantation which was observed until 20 months postoperatively. The results indicate that the progress of Parkinson's disease becomes slower and the clinical symptoms are less severe. L-dopa doses might be decreased which is followed by the reduction of its side effects. The opinions from different neurosurgical centers evaluating adrenal medulla autografts vary depending on the results noticed after those neurotransplantations. PMID- 8247245 TI - [Hormonal disturbances in patients with multiple sclerosis]. AB - The results of certain tests for the relationship between the hormonal system and the immune system may shed some light on the aetiology of autoimmune diseases, including multiple sclerosis. In patients with that disease signs are found of various hormonal disturbances. It is supposed that they are caused mainly by hypothalamic lesions, but they may influence the course of the disease. The paper is trial of a broader presentation of these problems. PMID- 8247246 TI - [Thoracic radiation myelopathy]. AB - In a 62-year-old patient 8 month after radiotherapy for right pulmonary hilus carcinoma signs of thoracis cord involvement with ascending course from Th10 to Th7. Cerebrospinal fluid and myelogram were normal. The patient died after about 12 months. Radiation myelopathy was confirmed by neurohistological examination. The results of experimental studies are quoted which could predict the possibility of early diagnosis of radiation myelopathy by rising level of basic myelin protein in cerebrospinal fluid. PMID- 8247247 TI - [Diagnostic difficulties in a case of carcinomatous meningitis]. AB - A case of carcinomatous meningitis clinically suggesting an inflammatory process is described. The patient died unexpectedly of an intercurrent cause and the neuropathological examination allowed to recognize a early stage of carcinomatous infiltration of meninges. PMID- 8247248 TI - [Basilar migraine: case report]. AB - The authors describe a case of basilar migraine in 35-year old woman. The diagnostic examinations performed in accordance with our expectations did not show abnormalities. Like other authors we think that the symptoms are caused by disturbances of vascularisation in basilar artery area. PMID- 8247249 TI - [Caudal tumors as a rare cause of subarachnoid hemorrhage]. AB - A case of cauda tumour (neurolemmona) is reported in which the presenting symptom was subarachnoid haemorrhage. The investigations for suspected cerebral vascular malformation brought no results. Only after appearance of clinical signs of cauda damage vertebral canal investigations made possible the correct diagnosis and proper treatment. PMID- 8247250 TI - [Assessment of the myorelaxant Sirdalud (produced by Sandoz company) on pain intensity in acute radicular syndromes]. PMID- 8247251 TI - [Report on the International Symposium on epidemiology of multiple sclerosis in Europe, Darmstadt (Germany)]. PMID- 8247252 TI - [Report on the VIIth Day of Child Neurosurgery: neurosurgical treatment of consequences of neuroinfections, Poznan (Poland)]. PMID- 8247253 TI - Current bibliographies of neuropeptides prepared by the University of Sheffield Biomedical Information Service. PMID- 8247254 TI - The effect of salt-loading on corticotropin releasing hormone and arginine vasopressin mRNA levels in the mouse hypothalamus: a quantitative in situ hybridization analysis. AB - Previously, we showed that during salt-loading in mice there was an acute rise in plasma ACTH levels after 2 days followed by a transient decrease after 4 and 9 days. Pro-opiomelanocortin (POMC) mRNA levels in the anterior pituitary increased after 2 days and returned to normal thereafter. In this study, changes in hypothalamic CRH and AVP mRNA levels during salt-loading were investigated using quantitative in situ hybridization histochemistry. CRH mRNA was expressed only in the paraventricular nucleus (PVN), while AVP mRNA was expressed in both the supraoptic (SON) and paraventricular nuclei. CRH mRNA levels were unchanged after 2 days salt-loading, but declined to 77% of control levels after 9 days. AVP mRNA levels rose to 260% and 634% of control levels in the SON, and to 352% and 522% of control levels in the PVN, after 2 and 9 days salt-loading, respectively. These data suggest a major role of AVP in the acute stimulation of ACTH secretion and POMC mRNA levels seen after 2 days salt-loading. Desensitization of AVP receptors at the corticotroph level and a centrally mediated inhibition of CRH release may account for the decrease of ACTH secretion and POMC mRNA levels in the anterior pituitary with prolonged salt-loading. PMID- 8247255 TI - Hypoalgesic action of bestatin analogues that inhibit central aminopeptidases, but not neutral endopeptidase. AB - Two analogues of the aminopeptidase inhibitor bestatin, Z 4212 (N-[(2S, 3R)-3 Amino-2-hydroxy-4-(4-methylsulphonyl-phenyl)-1-oxobutyl]-1- aminocyclopentanecarboxylic) and Z 1796 ((2S)-N-[(2S,3R)-3-Amino-2-hydroxy-4-(4 methylsulphonyl-phenyl)-1- oxobutyl]-L-leucine) were found to behave as hypoalgesics when intracerebroventricularly (i.c.v.) administered to mice in the hot-plate test. At high doses, Z 4212 was also found to reduce the pain threshold after intraventricular (i.v.) administration. Hypoalgesia induced by bestatin analogues was prevented by prior treatment with the opiate receptor blocker naloxone. Thiorphan, a potent inhibitor of NEP, was found to enhance the hypoalgesic effect of low doses of either Z 4212 or Z 1796. These results indicate that both the major opioid-degrading peptidases, i.e. aminopeptidases and neutral endopeptidase (NEP), are individually implicated in the hypoalgesia induced by peptidase inhibitors. In vitro studies showed that these new bestatin analogues readily inhibit aminopeptidases in membranes from mouse c. striatum whereas more than 1000 times the concentration was required for NEP to be blocked. Ex vivo experiments showed that, at variance with bestatin, the hypoalgesic action of Z 4212 or Z 1796 appeared to implicate central aminopeptidases but not NEP, so partially sparing the metabolism of other NEP substrates that might produce additional alterations (substance P and ANP). On the basis of the antitumour and immunomodulatory actions of bestatin, these new analogues might be potentially useful as mixed antitumour and hypoalgesic agents in malignancy. PMID- 8247256 TI - Binding in vivo of selective mu and delta opioid agonists: localization by autoradiography. AB - The in vivo binding properties of cerebral opioid receptors were investigated in mice after intracerebroventricular (i.c.v.) injections of iodinated FK33-824 and [D.Ala2]deltorphin-I which behave in vitro as highly selective ligands possessing high affinity for mu and delta receptors, respectively. [125I]FK33-824 and [125I][D.Ala2] deltorphin-I exhibited similar diffusion kinetics after i.c.v. injection and bound specifically to sites characterized pharmacologically as mu and delta receptors respectively. Autoradiographic analysis revealed that after i.c.v. administration, concentrations of [125I]FK33-824 and [125I][D.Ala2]deltorphin-I remained higher in the circumventricular than in the deep structure of the brain and that specific sites labelled in vivo were differently distributed from those observed after in vitro labelling. FK33-824 was 250 times more analgesic than [D.Ala2]deltorphin-I in the tail-flick test and at doses producing a similar analgesia, [D.Ala2]deltorphin-I occupied a high proportion of mu receptors. Furthermore, analgesic effect of [D.Ala2]deltorphin-I was antagonized by pretreatment with naltrexone but not by naltrindole, a selective antagonist of delta-opioid receptors. These experiments reveal the localization of mu and delta opioid receptors reached after i.c.v. injection and provide evidence to support the suggestion that delta-opioid receptors contribute little or none to the supraspinal antinociception. PMID- 8247257 TI - Met-enkephalin induced escape from dexamethasone immunosuppression. AB - The present study was undertaken to examine whether and what type of interaction occurs between a synthetic glucocorticoid, dexamethasone (DEX) and an opioid peptide, met-enkephalin (MENK) upon superoxide anion (O2-) release from human polymorphonuclear cells (PMN). MENK (10(-8) M) abolished suppressed O2- release from PMNs treated with 10(-7) M DEX. This was the case in unstimulated but not in PMNs stimulated with PMA. The effect of MENK was mediated through pertussis-toxin (PTX) sensitive G-protein and since it was abolished by H7 probably involves protein kinase C (PKC) as a second messenger system. Thus, MENK can abolish DEX induced suppression of O2- release from human PMNs possibly through the interaction of second messenger pathways. PMID- 8247258 TI - Immunomodulatory properties of diazepam-binding inhibitor: effect on human interleukin-6 secretion, lymphocyte proliferation and natural killer cell activity in vitro. AB - We have examined the influence of diazepam binding inhibitor (octadecaneuro peptide, DBI33-50) on cell mediated immune responses including LPS-stimulated monocyte IL-6 secretion, PHA induced lymphocyte proliferation and NK cell function in humans. All studies were performed in vitro on isolated human peripheral blood mononuclear cells in the absence or presence of synthetic DBI33 50. It has been shown that DBI33-50, in concentration between 10(-6)-10(-8) M, enhances the LPS-induced secretion of IL-6, as determined by specific bioassay for this monokine. On the other hand DBI33-50 (10(-6)-10(-12) M), had no significant effect on either PHA-induced lymphocyte proliferation or NK cell function. This data suggests a possible immunomodulatory role for DBI33-50 as an endogenous neuropeptide, which stimulates IL-6 secretion by human monocytes. PMID- 8247259 TI - Axon terminal P2-purinoceptors in feedback control of sympathetic transmitter release. AB - Extracellular ATP acts on P2-purinoceptors of peripheral effector cells, and this is the basis for its function as a (co-)transmitter in peripheral efferent neurons. ATP also acts on P2-receptors of neuronal cell bodies or dendrites, and this is the basis for its function as a fast excitatory transmitter at neuroneural synapses. A third site of action is axon terminals. In the vas deferens of the mouse, noradrenaline and ATP are postganglionic sympathetic co transmitters, and exogenous ATP acts on P2-purinoceptors of the sympathetic terminals to inhibit release of noradrenaline. Here we show that two P2 antagonists, suramin and Reactive Blue 2, increase the release of noradrenaline in mouse vas deferens. The increase is only obtained when there has been preceding nerve activity and is largely independent of the postjunctional response. These findings indicate a physiological function for axon terminal P2 purinoceptors: they mediate a novel prejunctional negative feedback in which released ATP inhibits subsequent transmitter release. PMID- 8247260 TI - Cultured GABA-immunoreactive neurons are resistant to toxicity induced by beta amyloid. AB - Neurodegeneration in Alzheimer's disease is characterized by a selective loss of particular cell populations. Several recent lines of evidence suggest that beta amyloid protein directly contributes to the disease's progression and is likely responsible for the observed pattern of neuronal death. We have previously demonstrated that aggregated beta-amyloid peptides are neurotoxic to cultured neurons. We now report that a neuronal population exhibiting GABA immunoreactivity is resistant to beta-amyloid-induced toxicity in vitro, a finding consistent with observations in the Alzheimer brain. Determination of the intrinsic neuronal characteristics responsible for resistance to beta-amyloid may prove beneficial in both understanding the mechanism(s) of beta-amyloid neurotoxicity and halting the disease's progressive neuronal degeneration. PMID- 8247261 TI - On the quantal analysis of hippocampal long-term potentiation and related phenomena of synaptic plasticity. PMID- 8247262 TI - Relative loss of the striatal striosome compartment, defined by calbindin-D28k immunostaining, following developmental hypoxic-ischemic injury. AB - The striatum is especially vulnerable to hypoxic-ischemic injury, both in adulthood and during development. Striatal injury is likely to play a major role in the chronic abnormalities of motor control which occur as a consequence of developmental hypoxia-ischemia. Previous studies have shown that two striatal neuron phenotypes, cholinergic and NADPH-diaphorase-positive, are resistant to developmental hypoxia-ischemia, but little is otherwise known of patterns of vulnerability among other striatal neurons. In particular, there has been no data available about patterns of vulnerability within the major striatal neuron group, the medium-sized neurons. Since a major anatomical and functional organization of these neurons is in their localization to either the striosome or the matrix compartments, we have examined the effect of developmental hypoxia-ischemia on these compartments using a quantitative morphologic analysis of immunostaining for the calcium-binding protein calbindin-D28k. We have found that there is a predominant loss of the striosome compartment; in the presence of a mean loss of 33% of total striatal area, there was a 49% decrease in striosomal area. There was also a 41% reduction in the number of striosomes, and a small (14%) but significant decrease in the mean area of individual striosomes. The striosome loss was uniform in the rostrocaudal dimension. At a cellular level, the density of calbindin-positive neurons, expressed as number per unit area, was preserved. While there are several possible explanations for the selective loss of the striosome compartment, one hypothesis is that the lower level of calbindin within these neurons makes them more vulnerable to increases in intracellular calcium, which has been postulated to play a role in hypoxic-ischemic injury. The predominant loss of the striosome compartment following hypoxic-ischemic injury may lead to an imbalance with the functionally distinct matrix system. Such an imbalance may contribute to the abnormalities of motor control observed after this form of injury. PMID- 8247263 TI - Selective neuronal death in the contralateral hippocampus following unilateral kainate injections into the CA3 subfield. AB - Intracerebral or intraperitoneal injections of kainic acid, an agonist at a class of glutamate receptors, have been extensively used to model temporal lobe epilepsy. In the present study we compared the types and distributions of selectively vulnerable neurons in the ipsi- and contralateral hippocampi following unilateral kainate injections into the CA3 subfield in order to examine whether "proximal" or "distant" neuronal damage resembled the pathology, and possibly also the mechanism, of human temporal lobe epilepsy. The degeneration of principal cells in the different hippocampal subfields was visualized by silver impregnation, and the loss of various types of non-principal cells was studied by immunostaining for the calcium binding proteins parvalbumin, calbindin-D28k and calretinin, as well as for somatostatin. In the first series of experiments various concentrations (ranging from 0.1 to 1 mg/ml) and volumes (0.5-2 microliters) of kainate were tested to induce reproducible damage in the contralateral hippocampus. The optimal dose, employed in the subsequent vulnerability studies, was found to be 3 x 0.5-microliter injections (over a period of 10 min) of a concentration of 0.33 mg/ml under ether anaesthesia, which was discontinued immediately after injection. Anaesthesia with equithesin was found to prevent contralateral cell death. Most if not all pyramidal cells in the CA3 region degenerated on the ipsilateral side, whereas the dentate granule cells, and the majority of CA1 pyramidal cells were resistant. A strikingly different pattern was found on the contralateral side, where CA1 pyramidal cells were almost completely lost, but the CA3 region (with the exception of CA3c) and the dentate gyrus remained intact. Three subpopulations of non-principal cells were found to be vulnerable in both hemispheres, the hilar somatostatin cells, spiny calretinin cells and mossy cells, as well as the spiny calretinin cells in stratum lucidum of CA3. The other subpopulations were resistant, except for those within the effective injection site. We propose that the "distant" (contralateral) damage resembles the pattern, and probably also the mechanism, of cell death in human temporal lobe epilepsy, whereas the ipsilateral damage does not. PMID- 8247264 TI - Differentiation of newly born neurons and glia in the dentate gyrus of the adult rat. AB - In order to determine whether newly born cells in the dentate gyrus of the adult rat express the neuronal marker, neuron-specific enolase, or the glial marker, glial fibrillary acidic protein, we performed combined immunohistochemistry and autoradiography on brains from adult rats perfused at various times ranging from 1 h to four weeks following [3H]thymidine administration. Light-microscopic examination revealed a negligible number of [3H]thymidine-labeled cells showing neuron-specific enolase immunoreactivity during mitosis. However, by two weeks after [3H]thymidine administration, a significant increase in the density of [3H]thymidine-labeled neuron-specific enolase-immunoreactive cells was detected. Three weeks following [3H]thymidine injection the majority of [3H]thymidine labeled cells (> 70%) were immunoreactive for the neuronal marker. At the four week time-point, [3H]thymidine-labeled neuron-specific enolase-immunoreactive cells were indistinguishable from neighboring granule cells. In contrast, glial fibrillary acidic protein immunoreactivity was observed in a small but significant number of [3H]thymidine cells at the 1-h time-point and the proportion of labeled cells that were immunoreactive for this cell marker did not increase with time. [3H]Thymidine-labeled cells that were immunoreactive for glial fibrillary acidic protein typically showed morphologic characteristics of radial glia at all time-points. At the 1-h time-point, the majority of [3H]thymidine-labeled cells were observed in the hilus (> 60%) with the remainder being located in the granule cell layer. However, with a four-week survival-time most [3H]thymidine-labeled cells (> 85%) were located in the granule cell layer. The majority of newly born cells in the adult dentate gyrus differentiate into neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247265 TI - Neural grafting to ischemic CA1 lesions in the rat hippocampus: an autoradiographic study. AB - Fetal hippocampal neurons were stereotaxically transplanted to five-day-old ischemic CA1 lesions in adult rat hippocampi. The recipient brains were examined 14 or 100 days later. The grafts survived well, and transplanted cells usually formed clusters in the host CA1 subfield. In vitro receptor autoradiography was employed to map the following receptors, the ligands indicated in parentheses being used for labeling: muscarinic cholinergic ([3H]quinuclidinyl benzilate), adenosine A1 ([3H]cyclohexyladenosine), kainate ([3H]kainic acid), spirodecanone ([3H]spiperone), opioid ([3H]naloxone), and GABAA ([3H]muscimol). The receptor autoradiographic technique showed significant binding of the six ligands in all hippocampal grafts two weeks after transplantation. One hundred days following transplantation, almost all receptors, especially muscarinic cholinergic, adenosine A1 and opioid receptor bindings in grafts, had significantly increased compared to bindings two weeks after transplantation. At this time, kainate and muscarinic cholinergic receptors in grafts had increased up to the near normal level of the CA1 in the hippocampus. Interestingly, adenosine A1 receptors in the grafted side had significantly increased not only in the CA1 but also in the stratum oriens of the CA3 compared with that in the non-grafted side. The increase of [3H]quinuclidinyl benzilate binding corresponded well with the innervation of acetylcholinesterase-positive fibers at 100 days after grafting. These results demonstrate that the transplanted neurons, which showed both pre- and post-synaptic autoradiographic markers in the ischemic CA1 lesions, are able to develop their properties and express the nature of normal hippocampal neurons. PMID- 8247266 TI - Excitatory and inhibitory synaptic processing in the accessory olfactory system of the female rat. AB - The accessory olfactory system constitutes a sensory system specifically involved in regulating neuroendocrine function and reproductive behavior. The chemosensitive structure of this system, the vomeronasal organ, exclusively innervates the accessory olfactory bulb, which in turn projects via mono- or disynaptic pathways to a limited number of regions implicated in endocrine and sexual function, including the amygdala and hypothalamus. The present study investigated synaptic processing between the different levels of this sensory system, with particular focus on the input from the accessory olfactory bulb to the medial amygdala and the reciprocal connections between the medial amygdala and the ventromedial hypothalamus. Extracellular single-unit recordings were obtained from medial amygdala neurons in anesthetized female rats to study the synaptic responses elicited by stimulation of the olfactory bulb and hypothalamus. Locally applied agonists and antagonists of amino acid neurotransmitters were tested for their ability to mimic and block these synaptic responses in an attempt to identify the neurotransmitters involved in the stimulated pathways. Electrical stimulation of the accessory olfactory bulb induced orthodromic responses in 88% of 288 medial amygdala neurons, with 67% showing orthodromic inhibition and 21% showing excitation as the shortest-latency response. Many cells displayed multiphasic responses with both orthodromic excitation and inhibition. In separate experiments, main olfactory bulb stimulation also produced excitatory (39%) or inhibitory (50%) orthodromic responses in medial amygdala neurons (n = 105), but the latency and distribution of responses were significantly different than with accessory olfactory bulb stimulation. Stimulation of the ventromedial hypothalamus also had prominent excitatory (29%) or inhibitory (59%) orthodromic actions on the majority of amygdala neurons. Out of 288 cells tested with both accessory olfactory bulb and ventromedial hypothalamus stimulation, 79% were orthodromically responsive to both areas, whereas only 3% responded to neither area. Fourteen (6%) medial amygdala neurons were antidromically activated from the ventromedial hypothalamus, and 11 of these 14 cells were simultaneously orthodromically responsive to accessory olfactory bulb stimulation, indicating that the accessory olfactory bulb can influence medial amygdala neurons that project directly to the hypothalamus. Iontophoretic application of drugs to the medial amygdala revealed that glutamate and GABA consistently mimicked the orthodromic excitatory and inhibitory responses, respectively, from both the accessory olfactory bulb and ventromedial hypothalamus. In some cases, ejection of bicuculline could block or reverse excitation of the orthodromic inhibitory responses, whereas kynurenic acid could eliminate the orthodromic excitation, indicating that excitatory and inhibitory amino acid transmitters are at least partially involved in these pathways.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8247267 TI - Rectospinal neurons: cell bodies, pathways, immunocytochemistry and ultrastructure. AB - A novel class of enteric neurons projecting directly from the rectal wall to the spinal cord, "rectospinal neurons", was investigated in rats by combined retrograde neuronal tracing, immunocytochemistry and electron microscopy. Rectospinal neurons were almost confined to myenteric ganglia of the distal rectum below the pelvic diaphragm and were labeled preferentially by injections into spinal cord segments L6/S1. Injections into more rostral spinal cord segments resulted in hardly any labeled enteric neurons. Dorsal and ventral rhizotomy experiments indicated an almost exclusive projection of rectospinal neurons through dorsal roots L6/S1 to the respective spinal cord segments. Among various peptides immunostained, vasoactive intestinal polypeptide and calcitonin gene-related peptide were selectively found in rectospinal neurons, which were also shown to contain calbindin, neurofilament protein- and peripherin immunoreactivity. Vasoactive intestinal polypeptide- and calbindin-immunostaining were frequently co-localized in the same perikarya, while calcitonin gene-related peptide-immunoreactive rectospinal neurons probably represented a separate population. Neonatal capsaicin treatment did not significantly reduce the number of rectospinal neurons. Electron microscopy revealed synaptic contacts on the surface of rectospinal neurons. Taken together, these results establish rectospinal neurons as an anatomically and neurochemically distinct class of enteric neurons. Synaptic contacts on rectospinal neurons suggest that these neurons may function as a direct link from the enteric to the central nervous system, thus indicating that connections between these two networks are reciprocal. PMID- 8247268 TI - Basic fibroblast growth factor promotes the survival of embryonic ventral mesencephalic dopaminergic neurons--II. Effects on nigral transplants in vivo. AB - The clinical potential of transplants of fetal dopaminergic neurons is limited by the fact that the percentage of cells surviving in such grafts is in general quite low. This report investigates the use of basic fibroblast growth factor administration (given either as a pretreatment or by repeated intrastriatal infusions) to promote the survival and behavioural efficacy of embryonic dopamine rich nigral transplants in rats. Pretreatment of the graft tissue by brief incubation with basic fibroblast growth factor increased the survival of tyrosine hydroxylase-immunoreactive (presumed dopaminergic) neurons in the grafts in comparison to control grafts, and accelerated the recovery in the transplanted animals in tests of drug-induced rotational asymmetry. However, the clear advantage seen in the rotation test conducted three weeks after transplantation had disappeared by nine weeks. The moderate effects of pretreatment were markedly enhanced by repeated intrastriatal infusion of basic fibroblast growth factor into the host animals over 20 days following transplantation. This resulted in > 100% increase in the number of dopaminergic neurons surviving in the grafts, and was accompanied by a significantly greater recovery of the rats' rotational asymmetries which persisted over the full nine weeks of testing. However, the repeated intracerebral infusions induced an inflammatory reaction in the striatum, and the associated trauma both complicates the interpretation of the mechanism of observed recovery and compromises the utility of this route of basic fibroblast growth factor administration for promoting graft survival. PMID- 8247269 TI - Midbrain dopaminergic neurons from postnatal rat in long-term primary culture. AB - Midbrain dopamine neurons project extensively throughout the vertebrate forebrain and influence a wide variety of brain functions. These neurons, which are believed to form a major brain reward system, are involved in initiation and control of motor programs, addictive behaviors, and determination of mood. Given their critical role in behavioral function, relatively little is known about their fundamental cellular physiological and pharmacological properties. A long term dissociated culture system for postnatal rat dopamine neurons was developed to permit both acute and chronic studies of these cells. Dopamine neurons were dissociated from slices of ventral midbrain from neonatal rat pups and maintained in cell culture for several months. The dopaminergic phenotype was confirmed by catecholamine fluorescence and by tyrosine hydroxylase immunocytochemistry. After four weeks in culture, dopamine neurons had cell bodies 10-40 microns in diameter, displayed either fusiform or multipolar morphology, and had processes with varicosities of 0.5-2 microns in diameter. Electrophysiological recordings were made from 71 dopamine neurons identified by 5,7-dihydroxytryptamine fluorescence after six to 67 days in culture. The neurons had resting potentials of -51 +/- 5 mV, broad action potentials with durations of 2.9 +/- 1.3 ms, and the majority of the neurons (65%) displayed anomalous rectification. Most dopamine neurons in culture fired spontaneously in a pacemaker-like manner with a frequency of 2.3 +/- 1.3 Hz, or in a bursting pattern, typically having two to seven action potentials per burst. All neurons tested had glutamate and gamma aminobutyric acid receptors, and 90% of neurons responded to dopamine or quinpirole with inhibition of firing, suggesting the presence of dopamine autoreceptors. Some neurons were inhibited by concentrations of quinpirole as low as 10 nM. The results show that midbrain dopamine neurons can be maintained in dissociated cell culture for periods of several months. These neurons can be identified prior to electrophysiological recording, and they express many of the physiological characteristics that have been reported for midbrain dopamine neurons in vivo. PMID- 8247270 TI - Differential vulnerability in the hindbrain neurons and local cerebral blood flow during bilateral vertebral occlusion in gerbils. AB - Differential vulnerability in the hindbrain neurons was examined immunohistochemically during hindbrain ischemia in the gerbil. Hindbrain ischemia was produced by extracranial occlusion of the bilateral vertebral arteries just before their entry into the transverse foramen of the cervical vertebra. Local cerebral blood flow was measured by quantitative autoradiographic technique after 5 min of ischemia and was reduced to less than 5 ml/100 g per min in the cerebellum, the pons, and the medulla, indicating that severe and reproducible hindbrain ischemia was induced immediately after occlusion. For immunohistochemical investigation, four gerbils each were used for each ischemic period of 5, 10, 15, and 30 min. Immunohistochemical lesions, detected by the reaction for microtubule-associated protein 2, were visible in the lateral vestibular nucleus and the cerebellar interpositus nucleus even after 5 min of ischemia. These results suggested that these areas were more vulnerable than others, although blood flow was markedly reduced in various regions of the hindbrain. In contrast, areas related to respiratory or cardiovascular control were rather resistant to ischemia. The present study suggests that selective vulnerability during hindbrain ischemia depends mainly on different metabolic characteristics inherent to various neurons in the hindbrain. PMID- 8247271 TI - A role for the nicotinic alpha-bungarotoxin receptor in neurite outgrowth in PC12 cells. AB - The addition of nicotine decreased neuritic outgrowth in PC12 cells in culture. This effect occurs as early as one day after addition of nicotine to the culture medium in a concentration-dependent manner. The nicotine-induced decline in neurite outgrowth was prevented by d-tubocurarine (10(-4) M) indicating that the effect was mediated through a nicotinic receptor. alpha-Bungarotoxin (10(-8) M) was also able to inhibit the nicotine-induced decrease in process formation in a dose-dependent manner. The concentrations of alpha-bungarotoxin required to affect process outgrowth correlated with those required to inhibit radiolabelled alpha-bungarotoxin binding. alpha-Bungarotoxin had no effect on [3H]noradrenaline release, a functional response mediated through the alpha-bungarotoxin insensitive neuronal nicotinic acetylcholine receptor, suggesting that alpha bungarotoxin specifically interacts with the neuronal alpha-bungarotoxin receptor. The present results suggest a functional role for the neuronal nicotinic alpha-bungarotoxin receptor in neurite outgrowth. PMID- 8247272 TI - Neonatal nerve injury causes long-term changes in growth and distribution of motoneuron dendrites in the rat. AB - Disruption of neuromuscular contact by nerve-crush during the early postnatal period results in the death of a large proportion of affected motoneurons. Increased activity and abnormal reflex responses are evident in those that survive. We have studied the aberrant dendritic morphology of surviving cells and have attempted to correlate the observed alterations in morphology with the above experimental findings. Motoneurons supplying the extensor hallucis longus muscles of the rat were retrogradely labelled with cholera toxin subunit-B conjugated to horseradish peroxidase. The dendritic tree of labelled cells was analysed in adult animals having undergone unilateral sciatic nerve-crush at birth. Unoperated control animals were also examined. Following nerve-crush at birth, total visible dendritic length was more than 30% smaller than control cells in the transverse plane. This decrease was confined largely to the medially directed segments of the dendritic field and appeared to be due to a reduction in dendritic branching combined with a failure to achieve the correct branch length. There was no overall change in total visible dendritic length in the longitudinal plane, but a reorientation of dendrites in favour of rostrodorsal regions was observed. There was no alteration in dendritic length in cells contralateral to the nerve injury. These results show that nerve injury during early postnatal development produces lasting changes in the distribution of motoneuron dendrites. The localized nature of these changes may explain the altered activity and induced death of motoneurons seen after neonatal nerve-crush. PMID- 8247273 TI - Long-term ethanol consumption by rats: effect on acetylcholine release in vivo, choline acetyltransferase activity, and behavior. AB - The extent and duration of cholinergic hypofunction induced by long-term ethanol consumption was investigated in the rat. Ethanol (20% v/v) was administered to male adult Wistar rats as a sole source of fluid for three or six months. Control rats received tap water. The body weight, food and fluid intake in ethanol treated rats were lower than in control rats throughout the treatment. After three months of ethanol consumption, and one week withdrawal, acetylcholine release in freely moving rats, investigated by microdialysis technique coupled to high-performance liquid chromatography quantification, was significantly decreased by 57 and 32% in the hippocampus and cortex, respectively, while choline acetyltransferase activity was significantly decreased (-30%) only in the hippocampus. A complete recovery of choline acetyltransferase activity and acetylcholine release was found after four ethanol-free weeks. Conversely, after four weeks of withdrawal following six months of ethanol treatment, the recovery in acetylcholine release was not accompanied by that in choline acetyltransferase activity, which remained significantly lower than in control rats in both cortex and hippocampus. The ability of rats to negotiate active and passive avoidance conditioned response tasks, tested after four ethanol-free weeks, was strongly impaired in both three- and six-month ethanol-treated rats. In conclusion, our experiments demonstrate that the development of a long-lasting cholinergic hypofunction requires at least six months of ethanol administration. The hypofunction affects choline acetyltransferase activity and acetylcholine release differently, and undergoes a remarkable recovery. PMID- 8247274 TI - The selective activation of dorsal horn neurons by potassium stimulation of high threshold primary afferent neurons in vitro. AB - Intracellular recordings from neurons in the dorsal root ganglion and dorsal horn, in an in vitro spinal cord-dorsal root ganglion preparation, were used to investigate the role of large and small afferent fibers in the sensory synaptic transmission of the superficial dorsal horn. Raising the extracellular potassium concentration from 3.1 to 25-50 mM in the dorsal root ganglion compartment evoked a large amplitude depolarization and blocked action potentials in the large neurons of dorsal root ganglion, and it synaptically excited dorsal horn neurons. Excitatory postsynaptic potentials that were evoked by electrical stimulation of large myelinated fibers, but not those evoked by activation of small unmyelinated fibers, were blocked by the potassium treatment of the dorsal root. Tetrodotoxin (0.3-10 microM), when applied to the sensory neurons, abolished action potentials in large myelinated fibers but had no effect on the potassium-induced depolarization of the soma of large neurons of the dorsal root ganglion. Bath application of tetrodotoxin to the dorsal root ganglion blocked the postsynaptic potentials evoked in dorsal horn neurons by electrical stimulation of large fibers (stimulus intensity 10-20V, 0.02 ms) but failed to block postsynaptic potentials induced by electrical stimulation of slow fibers (stimulus intensity > 35 V, 0.5 ms). In addition, the tetrodotoxin failed to block the synaptic activation of dorsal horn neurons which was induced by the application of high potassium to sensory neurons. Capsaicin (10-100 microM, 10 s), applied to the sensory neurons, resulted in a prolonged synaptic activation of the dorsal horn neurons and a subsequent long lasting desensitization. During the period of capsaicin desensitization, synaptic activation of dorsal horn neurons by application of high potassium to the dorsal root ganglion and electrical stimulation of slow fibers was blocked. The opioid receptor agonist (D-Ala2, D Leu5)-enkephalinamide (1 microM), applied to the spinal cord slice, abolished the dorsal horn neuron excitation evoked by electrical or chemical activation of slow primary afferent fibers. These findings indicate that high concentrations of K+ applied to the dorsal root ganglia selectively activate a primary afferent input to the dorsal horn, which is capsaicin sensitive and tetrodotoxin resistant. PMID- 8247275 TI - Does neuromelanin contribute to the vulnerability of catecholaminergic neurons in monkeys intoxicated with MPTP? AB - The question has been raised as to whether neuromelanin, a by-product of catecholamine metabolism which accumulates during aging in primate midbrain neurons, contributes to the selective vulnerability of subgroups of dopaminergic neurons in Parkinson's disease. 1-Methyl-4-phenylpyridinium (MPP+) a metabolite of 1-methyl, 4-phenyl, 1,2,3,6-tetrahydropyridine (MPTP) is toxic to dopaminergic neurons, particularly in primates, producing a motor syndrome similar to that observed in Parkinson's disease. To test whether this neurotoxin preferentially affects melanized neurons, the survival of melanized and non-melanized catecholaminergic neurons was analysed after MPTP intoxication in the midbrain of the cynomolgus monkey (Macaca fascicularis). Experiments were performed on six animals chronically treated with MPTP (two were severely disabled, four moderately affected) and two age-matched control monkeys. Two populations of neurons were examined on regularly spaced sections throughout the midbrain: catecholaminergic neurons, identified by tyrosine hydroxylase immunohistochemistry and neuromelanin-containing neurons, visualized by Masson's method. The total number of neurons of each type was estimated in the different midbrain catecholaminergic cell groups using computer assisted image analysis. In the midbrains of control animals not all catecholaminergic neurons contained neuromelanin. The percentage of melanized neurons compared to the total population of tyrosine hydroxylase-positive neurons was high in the substantia nigra pars compacta (81.5%) and in the locus coeruleus (98%), intermediate in the substantia nigra pars lateralis (70%), in the catecholaminergic cell group A8 (50%), and in the ventral tegmental area (41.5%) and almost nil in the central gray substance. In MPTP-treated monkeys, the severity of the loss of catecholaminergic neurons was variable within the different midbrain cell groups, though of similar intensity in severely and mildly disabled monkeys. A relationship was found between the loss of dopaminergic neurons in the different mesencephalic cell groups of MPTP-intoxicated animals and the percentage of melanized neurons they normally contain (r = 0.98; P = 0.04). The percentage loss of catecholaminergic neurons in the locus coeruleus, the only noradrenergic cell group studied, was lower than expected from the correlation curve obtained for dopaminergic cell groups. Altogether, these findings indicate: (i) that dopaminergic neurons are more vulnerable to MPTP-toxicity than noradrenergic neurons; and (ii) that among dopaminergic neurons, those containing neuromelanin are more susceptible, indicating a possible role of neuromelanin in MPTP toxicity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8247276 TI - [Inguinal hernia surgery under local anesthesia. Technical note and immediate results]. AB - The authors report their experience on local anesthesia for inguinal hernia repair in 164 patients. They describe the neuroanatomy of the inguinal region as well as the technique of loco-regional anesthesia. The Shouldice repair was most frequently employed, followed by the Bassini repair. Local anesthesia was very well tolerated by the patients, sometimes with the addition of mild sedation. The postoperative course was definitely shortened by the use of local anesthesia, and all patients were able to take a regular diet and to walk few hours after surgery. PMID- 8247277 TI - [Inguinal hernia repair with marlex mesh in a preperitoneal site using the classical inguinal access]. AB - Following a brief review of the main method of prosthetic repair used for inguinal hernia: Lichtenstein, Stoppa, Rives, etc, the authors propose an original technique consisting in the placement of a marlex patch below the trasversalis fascia, in a pre-peritoneal site, using a classical inguinal access route. The prosthesis is fitted round the spermatic funicle and fixed with a single stitch to the pubis and with a few others, in resorbable material, to the posterior face of the trasversalis fascia. Above it, plastic surgery is performed to bring the triple stratum closer to the reflexio of the inguinal ligament. The prosthesis is kept in place by positive abdominal pressure. The method outlined here has been used in 71 cases of primary or recurrent inguinal hernia. Follow-up lasted for 1-24 months and showed the low morbidity (4.5%) due to sepsis of the surgical wound, which did not require reoperation, and only 1 case of recurrent hernia which was probably caused by the incorrect positioning of the prosthesis. PMID- 8247278 TI - [Surgical treatment of gynecomastia. Indications and methods]. AB - In this paper the authors present their experience in the surgical treatment of gynecomastia; the different surgical techniques used (liposuction, adenomammectomy, liposuction associated with adenomammectomy) are compared bilaterally 42 patients (37 patients) were affected of bilateral gynecomastia, 5 patients monolateral only (age 18-38 years) were treated in the Department of Plastic and Reconstructive Surgery-Ca' Granda Hospital of Milan, from 1985 to 1991. 12 patients were treated by the adenomammectomy technique; plain liposuction alone was used in 4 patients only. The associated method (liposuction and adenomammectomy) was performed on 26 patients. The results obtained suggest that the associated method is usually the most effective, because esthetic results are excellent and post-operative complications are very low. In selected cases however the other techniques should be preferred, and the criteria of selection are discussed. PMID- 8247279 TI - [Primary malignant tumors of the small intestine]. AB - The clinical records of 10 patients with small intestinal tumors during a 27 years period were reviewed. The patients, 5 men and 5 women ranged from 17 to 79 years old. There were 6 carcinomas (60%), 3 lymphomas (30%), and 1 sarcoma (10%). Tumors were most frequent in the distal part of the small intestine (70%). The most common complaints were pain (70%) and obstruction (40%). All patients had surgical treatment; curative resection was attempted in 70% of the cases. Prognosis is poor, the mean survival time was 20.4 months. PMID- 8247280 TI - [Surgical treatment of toxic megacolon]. AB - After an introduction on the various aetiological factors involved in toxic megacolon, its onset, medical treatment and indications to surgery, the paper describes personal experience with seven cases. The report highlights the importance of medical treatment which is recommended as the appropriate initial therapeutic approach. Then after a review of evolving surgical techniques, it is suggested , again on the basis of personal experience, that the surgical approach should be neither excessively aggressive (total proctocolectomy with permanent ileostomy) nor excessively conservative (multistoma decompression). According to this report, the operation of choice is currently subtotal colectomy plus ileostomy and sigmoidostomy which combine modest morbidity and mortality with the possibility of future reconstruction. PMID- 8247281 TI - [Splenectomy in HIV-positive patients]. PMID- 8247282 TI - [Acute intestinal infarction. Diagnostic and therapeutic problems]. AB - The authors report their experience in thirteen cases of mesenteric infarction. They emphasize the difficulty in diagnosing and treating this entity, considering also the impossibility in most centers of performing emergency selective angiography of the superior mesenteric artery (SMA) When feasible, mesenteric rivascularization should always be attempted, followed 24 hours later by a "second look" procedure. PMID- 8247283 TI - Idiopathic varicocele: a comparison between types of surgery based on the results of spermiogenesis. AB - Varicocele must be regarded of the utmost importance in determinating male infertility. Surgical indications should be considered of primary interest in order to achieve a qualitative and quantitative improvement in spermiogram parameters in the majority of the cases. Recent reports showed that it is not the severity of the lesions to determine the alterations in seminal parameters but the length of time of the varicocele. In our report we consider the results achieved on the spermiogram parameters in two groups of 25 patients surgical treated with different techniques. The first group was treated, in others Centers, by means of the traditional technique of ligature of the spermatic vein (Ivanissevich, Palomo). We treated all the patients of the second group (53 patients) with microsurgical technique of anastomosis pampiniform plexus saphenous vein, and we control the sepermiogram parameters in 25 patients. The first group showed an improvement of the spermiogram parameters after 6 months from the surgical treatment in the 64% of the cases, while in the second group the improvement was of the 92%. Moreover we registered the persistence of this improvement after 12/36 months from the operation time. On the basis of these observations we think that the microsurgical technique leads to the best and longest results on the most invalidating cause of this pathology. PMID- 8247284 TI - [The use of a synthetic film (Bioprocess) in abrasions and second degree burns. Preliminary study in the emergency room]. AB - This study analyzes the usage of Bioprocess in abrasions and second degree burns, observing advantages and disadvantages of this treatment. This clinical research shows an important control against pain, a good hemostatic effect, an apparent reduction of healing time, an excellent cicatrization quality. PMID- 8247285 TI - [Frequency of surgical wound infections in patients treated with a barrier procedure. Multicentric study]. AB - In 3088 patients submitted to various surgical procedures an open clinical trial was carried out to evaluate the rare of surgical wound infections (SWI). All of them were treated, before and after surgery, with a foam emulsion (Bioshield) considered, from a theoretical point of view, to be able decrease the bacterial contamination of the surgical wounds. The SWI rate was really low (congruent to 2.3%) and a favorable role played by Bioshield cannot be excluded, both minimizing the skin lesions of the surgeon's hands and reducing wound contamination by exogenous flora. PMID- 8247286 TI - [Rehabilitation problems following lateral cervical lymphadenectomy]. PMID- 8247287 TI - [Solitary presacral echinococcal cysts]. AB - A rare case of a solitary hydatid cyst located in the pelvis, which was responsible for neural pain due to the compression of the left ischiatic nerve is described. Diagnostic and pathogenic problems related to the localization of hydatic cysts without liver and lung involvement are discussed. PMID- 8247288 TI - [Middle lobe syndrome]. AB - The authors, based on their actual experience with four cases and using previously published articles, state the pathological and clinical features of the middle lobe syndrome. Considering the physio-pathological processes, the authors determine the therapeutic program: the first approach would be pharmacological, second, if necessary, surgical-seeking to be as conservative as possible. PMID- 8247289 TI - [Intestinal occlusion caused by internal hernia of the retrocaecal recess. Description of a case]. AB - The authors describe an acute intestinal occlusion due to a rare case of strangulated intra-abdominal hernia of the retrocaecal peritoneal recess. They underline the rarity of this pathology which is often treated in emergency without any possibility of preoperative diagnosis. Therefore, in order to carry out surgical therapy with good results, a correct knowledge of the anatomical features of these hernias is necessary. PMID- 8247290 TI - [Timing in disorders associated to tracheo-pulmonary pathology]. AB - Although a wide variety of extrathoracic pathologies may be associated to tracheo pulmonary lesions, this paper draws attention to tumour pathologies due to their frequency and the difficulties involved in their treatment in relation to the question of "whether to carry out treatment" as well as the importance of timing. This dual nature is particular true of malignant conditions, since the lung may be the site of primary tumours and metastasis. The most frequent associations reported in the literature are tumours of the upper airways-digestive tract and colon carcinoma. Although tumours of the upper airways-digestive tract and lung cancer are both primary and primary tumour must be given priority treatment and only when oncologically satisfactory results have been obtained and there is no evidence of metastasis in other sites, are lung metastases. As far as concerns tracheal pathologies, this often takes the form of a locoregional extension of tumours from nearby organs such as the thyroid gland. In these cases, it is preferable to carry out total thyroidectomy and tracheal resection using T-T anastomosis. PMID- 8247291 TI - [Pulmonary hamartoma]. AB - Pulmonary hamartoma is a rare benign tumor often found by chance. Only in 10% of cases are some calcifications like "pop corn". Fibrobroncho-scopy is help only in endobronchial forms; they don't reach 10% of cases. Computed Tomography and transparietal biopsy can be diagnostic. In the uncertain cases and in presence of symptoms, thoracotomy with exeresis of tumor is justified. PMID- 8247292 TI - [Simple, complete, and comminuted fractures of the sternum. Analysis of 14 cases with osteosynthesis]. AB - The authors report 14 cases of fractured sternum operated upon in thoracic surgery at United Hospital in Foggia. Most of the ostheosynthesis of the sternum were done with two Patellar set sutures. The follow-up revealed good stabilization of the thoracic cage and good alignment of the fractured site. Postoperative respiratory distress was never noted, neither was thoracotomy ever required. PMID- 8247293 TI - [Surgical staplers in surgical emergencies. Our experience]. AB - This study describes the use of staplers during surgical treatment performed in emergency from January 1980 to December 1991. For the different, possible operations, some technical notes are examined and results reported. The advantages and risks related to the use of these devices are analysed. PMID- 8247294 TI - [Splenectomy versus splenic salvage. Our experience]. AB - The authors describe their experience in the surgical treatment of splenic lesions, mostly traumatic. In recent years a resolutely conservative attitude has been turned to splenic salvage, thanks to late findings on its immunological and haematological functions. Against 59 splenectomies for III-V degree lesions, the authors report a case of splenic trauma (II degree) without surgical treatment, monitored with clinical, haematological and instrumental (echography) test. PMID- 8247295 TI - [Surgery of cancer of the right colon. Analysis of 265 cases]. AB - The analysis of 265 selected cases of right colon cancer treated over a long period of time allows a number of points to be raised. Attention is drawn to the lack of progress made in diagnostic techniques, but this is thought to be due primarily to the vagueness of symptoms and their scarse investigation rather than to limits imposed by instrumental techniques. Moreover, no real benefit would be produced by a more aggressive approach using visceral or lymphatic exeresis. An earlier diagnosis of tumours due to differentiation, slow growth and rare association with adenoma should allow more encouraging results to be achieved. PMID- 8247296 TI - [Gallstone ileus]. AB - Gallstone ileus is a rare condition whose diagnosis may be difficult. From January 1976 to December 1991 4 cases of gallstone ileus were treated in our hospital, representing 1% of all patients admitted with mechanical bowel obstruction during that period. Three patients were treated by enterolithotomy alone to remove the impacted stone. In one patient intestinal resection of the bowel tract was associated followed by end to end anastomosis. One patient died (33%) seven days after surgery for cardiopulmonary failure. In one patient the obstruction resolved with the passage of a stone per rectum. The Authors conclude that enterolithotomy alone should be the standard procedure for gallstone ileus. The repair of cholecyst-enteric fistula should be done later only if there are continuing or recurrent symptoms. PMID- 8247297 TI - [Modified radical mastectomy in local anesthesia]. AB - The authors report their experience on the use of local anesthesia for modified radical mastectomy. They have used this modality in five patients who could not undergo general anesthesia because of underlying severe medical illness and in a sixth one who adamantly refused general anesthesia. They describe in detail the technique used, stressing the importance of a thorough knowledge of regional neuroanatomy. There were no complications in their small series. The authors conclude by stating that the use of local anesthesia may expand the indications for "toilet mastectomy" to include patients in poor medical condition, unsuitable for general anesthesia. PMID- 8247298 TI - [Is the surgical treatment of spontaneous bacterial peritonitis still up-to date?]. AB - We describe a case of spontaneous bacterial peritonitis in a 53 year old man affected by cryptogenic micro-macronodular cirrhosis, portal hypertention, splenomegaly and hypersplenism, who was admitted with hepatic failure and septic shock and successfully treated with antibiotics (combination of clindamycin and netilmycin), surgical abdominal drainage and splenectomy. This case gave reason for a literature review and an update on the therapeutic options in these high risk patients, especially concerning the role of surgery. Spontaneous Bacterial Peritonitis (SBP) is defined as a bacterial infection of ascitic fluid in the absence of any septic focus. It is a typical life-threatening complication of hepatic cirrhosis with ascites. Mortality is very high and ranges from 75% to 97% of patients, due to septic shock and hepatic failure (hepatorenal syndrome, hepatic encephalopathy, gastrointestinal bleeding). Infection with a single organism is found in most cases. Gram negative bacilli are present in about 70% of cases and E. coli (less frequently Klebsiella, Serratia, Pseudomonas) is principally found. Gram positive cocchi comprise an additional 30% of cases. Anaerobic and microaerophilic organisms seem to be rare causes of SBP (2.7-6%); this finding is probably due to the intrinsic bacteriostatic activity of ascites, which contains high oxygen tension (70 mmHg) and is an inhospitable environment for bacteroides and Clostridia. The prevalent isolation of enteric organism suggest that the gut is the most frequent source of infection, even if the pathogenetic mechanism is not yet well known. The right treatment depends on differentiating primary (SBP) from secondary peritonitis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247299 TI - [Massive hemobilia caused by necrotic hemorrhagic cholecystitis. Report of a case]. AB - The authors report a case of massive hematobilia due to hemorrhagic cholecystitis. Hematobilia is a rare pathology which affects the biliary tract and gallbladder. The first authors to describe hematobilia defined it as a hemorrhage of the gastroenteric tract due to the communication of blood vessels with the intra and extra-hepatic biliary tract and in some rare cases to the communication of the branches of the cystic artery within the gallbladder wall. Sandblom, in particular, specified that bleeding must be within the biliary tract and not secondary to an enterobiliary fistula. In 55% of cases the pathogenesis of hematobilia is traumatic, whereas in the remaining 45% the cause may be attributed to a variety of pathologies. Trauma include both non-surgical and surgical traumas; in the first group the most frequent cause is hepatic trauma, although it is worth taking into account the presence of post-traumatic arteriobiliary fistulas, lesions of arterial vessel walls with subsequent necrosis and rupture within the biliary vessels. Surgical traumas comprise lesions caused by therapeutic or diagnostic transparenchymal manoeuvres (PTC, biopsy). Non-traumatic causes include pathologies of vascular, cholecystic, inflammatory-infective and neoplastic origin. Symptoms are varied and take the form of anemia, massive bleeding with the onset of jaundice and pain in the hypochondrium and sometimes the epigastrium, whereas enterorrhagia is manifested by melena and more rarely hematemesis. The diagnosis must be made as quickly as possible; mortality increases with the delay in controlling hemorrhage. Differential diagnosis must take into account other causes of enterorrhagia, obstructive jaundice and anemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247300 TI - [Digestive hemorrhage due to an unusual cause: duodenal varices]. AB - The paper reports a case of digestive hemorrhage due to duodenal varices in a cirrhotic patient. The authors emphasise the extreme rarity and severity of this pathology and, in accordance with the literature, underline the difficulty of selecting the correct approach. PMID- 8247301 TI - [Appendicular mucocele. Clinical cases and review of the literature]. AB - The authors report three cases of mucocele of the appendix. They review the literature on this argument and discuss the hypotheses on etiology, pathologic and clinical classification, problems in diagnosis, principles of therapy and prognosis. PMID- 8247302 TI - [Partial resection of the spleen for echinococcus cysts with ultrasonic surgical aspirator]. AB - The superior extremity of the spleen was resected in a 52-year-old male patient due to a calcified echinococcus cyst, using an ultrasonic surgical aspirator (Cavitron). Blood loss was limited and no postoperative morbidity was reported. This method appears to be useful in all cases in which it is possible to avoid splenectomy in order to reduce the risk of uncontrolled postoperative infections. PMID- 8247303 TI - [Role of the endothelium in the control of vascular motility: physiologic aspects and physiopathologic implications in cardiovascular diseases]. AB - It was recently demonstrated that vascular endothelium produces many substances which modulate the vascular tone by acting locally (endothelium derived relaxing factor, angiotensin II, endothelin, prostacyclin). The authors review the physiological regulatory role of these substances and their involvement in cardiovascular diseases. They also consider the interaction between the endothelium derived factors and some cardiovascular drugs (such as calcium antagonists and ACE-inhibitors) and also the mechanisms of the possible protective action of these drugs on the vascular wall. PMID- 8247304 TI - [Monitoring of heparin therapy]. AB - The purpose of this study is to evaluate if the prolongation of activated partial thromboplastin time (aPTT/seconds) adjusted > 1.5 times the control value is a good schedule for heparin monitoring therapy. The authors have constructed a reference curve for the prolongation evaluation of heparin plasma concentration (units/ml) from corresponding aPTT/seconds. In this way, 6 patients at end dialysis treatment showed that aPTT/seconds prolonged > 1.5 times the control value is equivalent to heparin plasma concentration lungh than the limit of safe therapeutic regimen. This methodology of investigation is easy to perform and allows a continuous monitoring of heparin therapy. PMID- 8247305 TI - [The hand in hematologic diseases]. AB - Radiography of the hand often reflects the picture of generalized diseases, affecting both the muscolo-skeletal system and the others. Some of the most common hematologic disorders may be detected in roentgenograms of the hand, especially in the anemias, but also in plasma cell dyscrasias and proliferative malignant diseases (i.e., lymphomas and leukemias). On the basis of their experience, the authors have reviewed and discussed the radiographic "pattern" of the hand in several hematologic conditions (i.e., anemias; thalassemias; sickle cell disease; lymphomas; multiple myeloma; etc.), and their pathogenesis. Radiographies of both the hands, in antero-posterior view, were performed using Kodak-Min R film; xeroradiography was performed--in the same projection--using Rank Xerox plate, developed always in "positive mode" in 125 Rank Xerox System, conditioned with contrast "D", for emphasizing osseous details. Recently, digital radiography--employing PCR system--has substituted xeroradiography, because of its well-known properties and diagnostic advantages: in this manner, changes in bone and soft tissue are demonstrated on the same image, with augmentation of diagnostic information, with reduced dose to patient. In our series, hand is always involved (100% of cases) in thalassemias: lesions are characterized by diffuse osteopenia (washed out melted appearance), with widening of bone marrow space, diaphyseal convex aspect of the long bones, thinning of the cortex, and cyst-like changes (rain drops). Lesions disappear completely after the hypertransfusion regimen (HTR). Following chelation therapy, lesions of the wrist and hand are similar to those described in rickets and/or scurvy. Sometimes, the hand is characteristically affected in sickle-cell disease--particularly in the so called hand-foot syndrome--as "cone-deformity". In multiple myeloma incidence of hand involvement is 2.9%: lesions reflect general abnormalities observed in other skeletal sites, and they consist in multiple well-circumscribed lytic lesions. In amyloidosis, poorly defined radiolucent areas may be discovered. In non-Hodgkin lymphoma, incidence of hand involvement is less frequent, approximately 0.2%: the radiographic pattern is aspecific (mottled lytic lesions), sometimes simulating multiple myeloma and/or leukemias. In hemophilia, swelling of soft tissues, around the interphalangeal joint, related to intra articular and/or per-articular hematoma, is observed. The other conditions reflect general radiographic features of anemias, which are of three main types: 1. the over active marrow (i.e., polycythemia); 2. the infarction of bone (i.e., sickle-cell disease); 3. non-specific findings, resulting from chronic illness (delays of maturation; dwarfism; osteopenia; tendency to infection). PMID- 8247306 TI - [Occult aneurysms of the abdominal aorta. Our echographic experience]. AB - The authors report their experience in evaluating abdominal aorta aneurysms (AAA) by ultrasound examination. The incidence of this kind of disease is growing up in the last years and an earlier diagnosis increases the surviving possibilities of the patient affected, above all if symptomless, leading to a correct treatment: a semestral or annual follow-up of the aneurysms with a diameter smaller than 5 cm and surgery of the aneurysms with a diameter wider than 5 cm, because of high risk of rupture. The authors stress the importance of extending abdominal ultrasound examination to the aorta in all patients over 50 year old especially if affected by vascular diseases, hypertension and diabetes. In their experience the authors found 106 AAA; 61% of them with less than 4 cm diameter. PMID- 8247307 TI - [The oral strontium load test in the assessment of intestinal calcium absorption]. AB - A close correlation between calcium and strontium intestinal absorption has been described. In this study, a test using Stable Strontium has been assessed in women without abnormal calcium or bone metabolism, with no history of drugs which might affect calcium or bone metabolism. Decreasing values of Strontium intestinal absorption, according to the length of the postmenopausal period, have been observed. Besides, the Stable Strontium Test has been given in postmenopausal women with osteoporotic femur fractures. In comparison with age matched healthy women, this latter group showed a significantly lower Strontium intestinal absorption. Analogous behaviour has been reported for Calcium intestinal absorption. PMID- 8247308 TI - [Antihypertensive action of nicardipine retard in 24 hours and its effect on stress]. AB - Thirty-six patients (17 males and 19 females), aged between 40 and 70 years old (mean age 55.9), suffering from slight or moderate arterial hypertension, were monitored for four weeks after 14 days of placebo treatment. In a double-blind and random study 24 patients were treated with Nicardipine Retard (40 mg twice a day) whereas a further 12 received placebo twice a day. Sphigomanometric controls carried out after two and four weeks showed a significant reduction in arterial pressure only in those patients receiving active treatment. 24-hour out-patient monitoring of arterial pressure, carried out using Spacelabs 5300, showed a reduction in both systolic and diastolic arterial pressure throughout the day in subjects treated with calcium-antagonists compared to the placebo group. The normal physiological 24-hour trend of arterial pressure was always taken into account. The pressure response to a cold pressor test, mental arithmetic test, isometric and dynamic effort tests, measuring using a cycloergometer, was not modified by anti-hypertensive treatment, thus confirming the preservation of normal physiological behaviour during daily activities. There was no significant change in heart rate and the drug was well tolerated. PMID- 8247309 TI - [Weekly low-dose methotrexate in rheumatoid arthritis. Review of the literature]. AB - Methotrexate (MTX) is an antifolic drug that in recent years has been largely employed in the treatment of Rheumatoid Arthritis (RA). Both short and long term clinical trials have demonstrated its efficacy and good tolerability. It induces a significant improvement of all clinical variables and a decrease in the erythrocyte sedimentation rate and other acute phase reactants with a steroid sparing effect. The probability of continuing MTX therapy for up to 5 years is 46 55% whereas that of continuing gold, hydroxychloroquine, sulfasalazine or D penicillamine therapy is less than 20%. MTX is a rapidly acting drug with a clinical response within 4 weeks and a plateau phase after 6 months of therapy. Discontinuation of long-term MTX therapy induces a flare-up of the disease so that patients receiving long-term MTX must continue the drug to maintain clinical benefits. In spite of its clinical efficacy, MTX does not seem to have a significant effect on disease progression as determined radiographically. In this respect, MTX appears to have some superiority when compared to azathioprine, but not when compared to gold salts. MTX has been employed in patients with RA unresponsive to other Disease-Modifying Antirheumatic Drugs (DMARDs), but according to some recent views on the therapeutic strategy of RA, it could be used in early RA as a first choice drug. Toxic effects are the main reason in limiting long-term MTX treatment. Hepatic toxicity is one of the more common side effects of MTX, but the recognition of its "risk factors" such as alcohol abuse, may reduce it. Acute pneumonitis is one of the more severe complications of MTX therapy and may be life-threatening. In RA patients treated with MTX are also reported complications of immunosuppression, such as Pneumocystis carinii pneumonia whose clinical-radiological picture may be similar to that of acute pneumonitis. The mechanism of action of low-dose weekly MTX in RA is still unclear, but it might be more antiinflammatory than immunosuppressive, as supported by the rapid clinical response. The inhibition of Interleukin-1 activity or other inflammatory cytokines and inflammatory cells may play an important role in the antiinflammatory effect of MTX. MTX effects in RA are not fully understood and further studies are needed to clarify its mechanism of action and its place in the therapeutic strategy of this disease. PMID- 8247310 TI - [Adrenal cystic masses. Our experience]. AB - Cystic masses of the adrenal gland are clinically and pathologically rare findings and few cases have been reported up to now in the medical literature. In the present work 5 new cases are reported: 3 adrenal pseudocysts, 1 lymphangioma, 1 cystic pheochromocytoma. In 3 cases there were clinical symptoms of retroperitoneal mass (lumbar pain, palpable mass, digestive symptoms); in 3 cases conventional radiology was helpful; ultrasonography was used for diagnosis in 1, CT scan in 2. In the pheochromocytoma case the real nature of the mass was determined through fluid hormone determination after fine needle puncture. All cases were treated by surgery. PMID- 8247311 TI - [Diagnostic role of the pharmacologic stress with dipyridamole in reduced coronary reserve associated with CREST syndrome. Description of 2 cases]. AB - Myocardial lesions in subjects with CREST syndrome, may be a manifestation of focal ischemic injury resulting from functional vascular disease. We describe the case of women with angina in CREST syndrome in which only the pharmacological stress with dipyridamole was able to demonstrate signs of ischemia. PMID- 8247312 TI - [Oral desmopressin in the treatment of central diabetes insipidus]. PMID- 8247313 TI - [Obstetrics and neonatology in the Bible]. AB - Biblical quotations related to obstetrics and to the problems of newborn are reviewed and commented. So, we may have an idea on how such problems were understood at the time of composition of the Holy Bible. PMID- 8247314 TI - [Viral encephalitis transmitted by arthropods]. PMID- 8247315 TI - Effect of alcohol intake on lipids and fat-soluble vitamins in blood. AB - Several investigators have observed that a moderate intake of alcohol can have a positive effect on health (lowering the risk of cardiovascular disease, gall stones, etc.). Our objective was to investigate how some blood constituents are affected by the consumption of different levels of alcohol. Erythrocyte phospholipids and fatty acids composition and plasma fat-soluble vitamins were determined in 28 volunteers who were divided into three groups according to their alcohol intake: non-drinkers, moderate drinkers--consumption of 20-60 g/day, and heavy drinkers--more than 60 g/day. A relation was seen between alcohol consumption and vitamin status; furthermore alpha-tocopherol and beta-carotene were significantly lower in heavy drinkers. The erythrocyte phospholipid compositions of the non- and moderate drinkers showed no significant differences whereas a definite relation was noted when the alcohol intake was higher than 60 g/day. In conclusion it is possible to say that a moderate alcohol intake does not have any significant effect on the phospholipid and fatty acid composition; only the vitamin status is significantly influenced, but to confirm our results it is essential to evaluate other variables involved in this problem such as the lifestyle, physical activity and the eating habits of the subjects tested. PMID- 8247316 TI - [Frequency of hereditary neurologic diseases. A clinical study]. AB - INTRODUCTION: The nervous system is affected in 30% of hereditary monogenic disorders and as many as 500 single-gene disorders display major neurologic symptoms. We have studied the frequency of hereditary neurological diseases to assess their importance in daily hospital activity. Only single-gene hereditary diseases with central or peripheral nervous system involvement were considered; thus chromosomal diseases and diseases with multifactorial etiology were excluded. METHODS: We surveyed admission to in- and out-patient departments of Neurology, Pediatrics, and Dermatology of the Aosta Regional Hospital for the calendar years 1982-1991, collecting 229 cases, 95 women and 134 men. Out-patient departments held 126 patients, the others came from in-patient departments. Admission to the neurological in-patient department were 1.8% of total neurological admissions in the same period. Each diagnosis was assigned to the code number of the International Classification of Diseases (ICD-IX Revision, 1975). RESULTS: We found 33 different phenotypes. Most frequent diagnoses were: essential tremor (89 patients), hereditary sensory-motor neuropathy (HSMN) type I (28), Huntington's chorea (13), progressive muscular dystrophy limb-girdle type (8), neurofibromatosis type I (9), HSMN type II (9), spinocerebellar ataxia (9), hereditary spastic paraplegia (7), spinal muscular atrophy type IV (5), myotonic dystrophy (5), cerebellar ataxia (4), HSMN type III (4), spinal muscular atrophy type II and III (3), tuberous sclerosis (3). Essential tremor mostly affected persons in the over-50 age groups. On the contrary, the other neurologic monogenic diseases were diagnosed in all ages with the following age-group breakdown: 0-9, 11%; 10-19, 16%; 20-29, 15%; 30-39, 8%; 40-49, 11%; 50-59, 19%; 60-69, 14%, 70+, 7%. Consistently with the general rule, autosomic recessive diseases have the earliest onset and autosomic dominant ones the latest; HSMN, spinal muscular atrophy and Huntington's chorea were the disorders diagnosed in older age group. DISCUSSION: Although the frequency of the single neurologic monogenic disease is low (with the exception of essential tremor), their overall prevalence is higher than the prevalence of multiple sclerosis or the peripheral neuropathies. All age-groups are involved. We separated three groups of diseases: 1) two relatively benign diseases, essential tremor and the HSMN, affecting half of our patients; 2) five severe and more common diseases (Huntington's chorea, progressive muscular dystrophy limb-girdle type, neurofibromatosis type I, spinocerebellar ataxia, hereditary spastic paraplegia), affecting 1/4 of patients; and 3) a group of rare, mostly severe diseases, affecting the remaining 1/4 of patients. PMID- 8247317 TI - [Treatment with an aldose reductase inhibitor in peripheral neuropathy in elderly diabetic patients]. AB - Diabetic neuropathy (DN) is one of the most frequent and severe long-term complications in elderly diabetes. Most scientists believe that onset of chronic complications in diabetics is due to prolonged glycometabolic imbalance. Recently aldose reductase inhibitors (ARI) have been used in the treatment of DN. We studied 100 elderly subjects affected by diabetes mellitus who were treated with tolrestat, an ARI, for a year and a control group who received a placebo. All subjects underwent the following examinations at the start of treatment and then at 8, 24, 42 and 52 weeks: 1. electromyography of the lower and upper limbs', 2. biotensiometric evaluation of the vibratory perception threshold (VPT), 3. glycosylated hemoglobin, 4. hourly glycemic profile, azotemia, creatininemia, hemochrome, cholesterol, triglycerides, bilirubin, electrolytes, protidogram, urine. The patients on tolrestat showed: increased nerve conduction velocity (mean values 2.3 m/sec) of the ulnar nerve; increased nerve conduction velocity (mean values 3.9 m/sec) of the peroneal nerve; reduced VPT at the right and left first toe (mean values 5 and 7 volts respectively); reduced VPT at the right and left malleoli (mean values 10 and 8 volts respectively). The authors recommend prompt, long term ARI treatment be initiated before the onset of evident signs of neuropathy in elderly subjects. PMID- 8247318 TI - [Thrombotic thrombocytopenic purpura (Moschowitz syndrome). A clinical case and review of the literature]. AB - A case of young woman affected by relapsing Thrombotic Thrombocytopenic Purpura (TTP), probably beginning in September 1986 and relapsing after 6 years, in June 1992, is described. After a first Complete Remission due to Plasma Exchange and Antiplatelet Agents, the patient relapsed during maintenance therapy with Ticlopidine and, over 1 month, developed two relapses which were resistant to traditional therapy. Showing resistance even to Iloprost, the patient had a Complete Remission only after a 2nd IV bolus of Vincristine according to Gutterman's scheme (modified), with prolonged therapeutic wellbeing. Many authors say that Plasma Exchange and Vincristine therapy must first be used in TTP and tried in refractory TTP. This one experience of ours should confirm that. From this TTP case, the authors begin a wide review of the literature, examining history, etiopathogenetic and therapeutic debate, possible related diseases, differential diagnosis, evolution, prognosis and future developments of this mysterious and rare illness. PMID- 8247319 TI - Effect of one hyperbaric oxygen-induced convulsion on cortical polyamine content in two strains of mice. AB - In rat striatum, after one hyperbaric oxygen (HBO)-induced convulsion, polyamine changes are found that could promote N-methyl-D-aspartate (NMDA) activation. In the HBO-sensitive CD1 mouse, unlike in the common C57 strain, there is some support for NMDA activation after the HBO seizure. We measured PA cortical content before and after the first HBO-induced convulsion (about 608 kPa O2) in CD1 and C57 strains. Putrescine, spermidine and spermine were dansyl derived and analysed by HPLC. Exposure to HBO significantly increased putrescine content only in CD1 though a similar trend was observed in C57. No further increase was observed after convulsion whatever the strain. There were no significant changes in spermidine or spermine to support NMDA activation. Therefore, putrescine increase in CD1 cortex could reflect the free radical formation that is known to be greater in CD1 than in C57 mouse. Attempts to increase putrescine levels before HBO exposure hastened HBO-induced convulsion, less than spermidine or spermine. Because of physiological polyamine interconversion, additional experiments with indirect manipulation of putrescine levels and study of their time-course would precise these preliminary reports on putrescine and HBO. PMID- 8247320 TI - Parvalbumin as an anatomical marker for discrete subregions of the ambiguus complex in the rat. AB - The topography of parvalbumin-immunoreactive neurons within the ventrolateral medulla of rats was investigated. Parvalbumin is a member of the 'EF-hand' family of Ca-binding proteins and is present in certain cell types within the central nervous system (fast-firing neurons with high metabolic rates). Parvalbumin immunoreactive neurons were located in discrete rostrocaudal divisions of the ambiguus complex corresponding to regions containing respiratory-related neurons. Based on the location of physiologically characterized respiratory-related neurons reported in the literature, parvalbumin immunoreactivity does not appear to distinguish inspiratory- from expiratory-related neurons. PMID- 8247321 TI - Glutaminase immunoreactive neurons in the rat dorsal root ganglion contain calcitonin gene-related peptide (CGRP). AB - The co-localization of glutaminase and calcitonin gene-related peptide (CGRP) was examined with immunohistochemistry in the rat dorsal root ganglion (DRG). The majority of the DRG neurons were immunoreactive for glutaminase and all DRG neurons that contained CGRP also contained glutaminase. These results indicate that some DRG neurons release glutamate and CGRP from the same axon terminals in the spinal cord. Co-release of glutamate and CGRP from primary afferents may have multiple effects including fast and slow neurotransmission of sensory information in the spinal cord. PMID- 8247322 TI - Bacterial collagenase disrupts extracellular matrix and opens blood-brain barrier in rat. AB - Bacterial collagenase causes hemorrhagic necrosis of brain. We studied the enzyme's effect on blood-brain barrier (BBB) permeability and extracellular matrix (ECM) structure by radiolabeled tracers and electron microscopy. Adult rats had intracerebral injection of bacterial collagenase. Brain uptake from blood of [14C]sucrose was measured in 24 rats 0.5 h to 14 days after injection. 12 rats had ultrastructural studies 1 h after collagenase injection. Brain uptake of [14C]sucrose is maximally increased at 0.5 h, remaining significantly increased for 7 days. Ultrastructurally, some vessels had widening of basal lamina while others had severe disruption of basal lamina with stretching of endothelial cells. We conclude that bacterial collagenase disrupts ECM and opens BBB. PMID- 8247323 TI - Antibody to an abnormal protein in amyotrophic lateral sclerosis identifies Lewy body-like inclusions in ALS and Lewy bodies in Parkinson's disease. AB - Ubiquitinated cytoplasmic inclusions are a characteristic feature of the neuronal pathology of neurodegenerative diseases. Immunocytochemical techniques have identified intermediate filaments associated with ubiquitin-immunoreactive inclusions in Alzheimer's disease (AD), Parkinson's disease (PD), and Pick's disease; however, no core protein has been detected in the ubiquitinated inclusions in amyotrophic lateral sclerosis (ALS). The pathogenesis of these inclusions is not known, but the inclusion may result from an accumulation of an abnormal proteins. Here we report a novel protein of 32.5 kDa detected by polyacrylamide gel electrophoresis, in the spinal cord in ALS patients. A polyclonal antibody raised against this protein and used for Western blotting, suggests that the novel protein is related to actin. Immunocytochemical studies using this antibody indicate that the protein is found in Lewy body-like inclusions in anterior horn cells of ALS, and in Lewy bodies in the substantia nigra in PD. PMID- 8247324 TI - NADPH diaphorase innervation of the rat anococcygeus and retractor penis muscles. AB - NADPH diaphorase histochemistry was used to determine whether the rat anoocccygeus (AC) and retractor penis (RP) muscles are innervated by nerves capable of synthesizing nitric oxide. In both tissues, muscle fascicles were enclosed by a varicose plexus of NADPH diaphorase positive (ND+) fibers. Perikarya of neurons on the surface of the AC muscle were also intensely stained for NADPH diaphorase. Many AC-RP ganglion cells in the pelvic plexus, located by the retrograde tracer Fluorogold, also stained for the enzyme. However, a significant population of AC-RP neurons in this location remained unstained. These results provide further evidence that nitric oxide may be an important neurotransmitter in these tissues. PMID- 8247325 TI - A dose-response study of dextrorphan in permanent focal ischemia. AB - The dose-response curve and time window of efficacy for dextrorphan in permanent focal brain ischemia leading to infarction was studied in the rat. With pretreatment, the maximum effective dose of 20 mg/kg reduced the eventual infarct volume by greater than 50%. Delayed drug administration, up to 45 min following permanent middle cerebral artery occlusion, significantly reduced infarct size. PMID- 8247326 TI - Hippocampal mossy fiber changes in mice transgenic for the human copper-zinc superoxide dismutase gene. AB - The copper-zinc superoxide dismutase (SOD-1) gene, located on chromosome 21 and triplicated in Down's syndrome (DS), is suspected to be involved in the neuropathology observed in Alzheimer's disease (AD), DS and physiological aging. In order to explore the effect of an overproduction of SOD-1 in the mouse hippocampus, we investigated the Timm-stained mossy fiber (MF) innervation in the hippocampus of transgenic mice for the human SOD-1 gene (hSOD-1 mice). The results showed a decrease of the MF projection area in the hSOD-1 mice overexpressing the SOD-1 protein. These findings suggest that free radicals could play a role in this particular synaptic loss. PMID- 8247327 TI - [125I]galanin binding sites in the rat frontal lobe are guanine nucleotide sensitive and display a low regional index of occupancy. AB - As the neuropeptide galanin is possibly involved in spatial learning, we investigated both the precise location and the binding features of the receptors within the rat frontal cortex using quantitative autoradiography. Galanin receptors predominated in layers I and V of medial and lateral frontal cortex with a low regional index of occupancy by endogenous galanin. These receptors might be of functional relevance since the guanylnucleotide Gpp(NH)p inhibited [125I]galanin specific binding in each labeled region in the frontal lobe. Nevertheless, the striking areal difference of expression of this effect within the medial frontal cortex suggests that [125I]galanin binding sites might be coupled to one or more types of G-proteins related to the functionally distinct cortical subareas. PMID- 8247328 TI - The c-fos gene and early-onset familial Alzheimer's disease. AB - A gene for early-onset familial Alzheimer's disease (FAD) is located on chromosome 14q24.3. The c-fos gene (FOS) is also located in the same band of this chromosome and is thus a candidate for the FAD locus. A yeast artificial chromosome (YAC) clone was identified which contains FOS. This YAC also contains the short-tandem repeat polymorphic (STRP) locus D14S76, placing FOS in the FAD region between D14S53 and D14S43. No recombinants were observed between D14S76 and FAD, and a maximum positive LOD score of 7.20 at a recombination fraction of 0.001 was observed for linkage of this marker to FAD. DNA sequence analysis was used to examine FOS in two affected subjects from an FAD family in which the chromosome 14 FAD locus is clearly responsible for the disease. The coding regions and parts of the 5' and 3' untranslated sequences of FOS were sequenced; no FAD-related mutations were observed. This work suggests that the FOS gene is not the chromosome 14 FAD locus although we cannot exclude the possibility that a mutation in an as yet unknown regulatory region is responsible for the disease. A new polymorphism was detected in the third intron of the gene. PMID- 8247329 TI - Immunohistochemical demonstration of neuron specific antigen, HPC-1 in the enteric nervous system of the guinea-pig distal colon. AB - The HPC-1 antigen is a newly identified neuron specific membrane protein in the central nervous system. The HPC-1 antigen was revealed similarity to epimorphin. The presence of HPC-1 antigen in the enteric nervous system of guinea-pig distal colon was immunohistochemically demonstrated using the antibody against the HPC 1. Immunohistochemical study clearly revealed the topography and structure of the enteric nervous system of the guinea-pig distal colon. HPC-1 was present only in the nervous system and entirely distributed. HPC-1 antigen is present at the surfaces of ganglion cells, but not in the cytoplasm. PMID- 8247330 TI - The effects of acute cocaine administration on auditory event-related potentials in rats. AB - Cocaine administration has been shown to affect several sites in the limbic forebrain. The nucleus accumbens has been implicated as an important site for the reinforcing aspects of this drug whereas, the amygdala and hippocampus may be more involved in drug conditioning and/or drug induced changes in the perception of stimuli. In the present study, auditory event-related potentials (ERPs) were utilized to explore the effects of cocaine on sensory processing in several limbic sites. Eleven adult male Wistar rats were stereotaxically implanted with electrodes aimed at the dentate gyrus, dorsal hippocampus (CA1-CA2), amygdala, and nucleus accumbens. The rats received intraperitoneal injections of either saline, 2.5, 5.0, 10.0 or 20 mg/kg of cocaine. The ERPs were recorded in response to an auditory 'oddball' paradigm consisting of frequently and infrequently presented tones. Cocaine was found to produce a dose dependent decrease in the variance of the amplitude of several ERP components in the amygdala and the hippocampal formation but not in the nucleus accumbens. Cocaine also produced a decrease in the latency of the N1 component in the amygdala. No significant changes were observed in the amplitude of the ERP components following any of the cocaine doses studied, suggesting that cocaine may not increase the salience of auditory stimuli in these structures. The observed reduction in the variance of the ERPs may suggest an increased focussing of the animals' attention to the auditory stimuli. These studies also suggest that the sensory/cognitive aspects involved in cocaine's actions, as quantified by ERPs, may primarily involve the hippocampal formation and the amygdala, but not the nucleus accumbens. PMID- 8247331 TI - Pharmacological characteristics of GABAA responses in postnatal suprachiasmatic neurons in culture. AB - The suprachiasmatic nucleus (SCN) is considered to be an endogenous circadian pacemaker. Previous studies have suggested functional roles of gamma-aminobutyric acid (GABA) in the control of circadian rhythms. In this study, the responses to applied GABA in cultured SCN neurons dissociated from postnatal rat hypothalamus were investigated using whole-cell voltage-clamp techniques. GABA and muscimol induced a large current response (EC50 values 5.3 and 1.6 microM, respectively), which was blocked by the GABAA antagonist bicuculline. This current response was also blocked by Zn2+ (0.5-50 microM) in a concentration-dependent manner, but was not potentiated by diazepam (10 microM) or ethanol (21 mM). These characteristics seem to correspond to those of GABAA receptors that lack gamma-type subunits. PMID- 8247332 TI - Differential action of phenytoin and carbamazepine on cortical interhemispheric responses in young but not in adult rats. AB - Single as well as paired responses to stimulation of the contralateral sensorimotor cortex were recorded in adult and 12-day-old rats under urethane anesthesia. Carbamazepine (50 mg/kg i.p.) prolonged the peak latencies of single responses in both adult and young rats, while phenytoin (60 mg/kg i.p.) prolonged the latencies in adult animals but shortened them in the young ones. Excitability cycles of the amplitude of the initial positive-negative complex remained uninfluenced by either drug in both age groups. Since solvent alone did not induce any effects in either test, our results demonstrate a qualitative change in the action of phenytoin during development. PMID- 8247333 TI - Histamine-containing nerve fibers innervate human cerebellum. AB - Histamine is found in nerve cell bodies of the tuberomammillary nucleus in mammalian brain. This nucleus is prominent in human brain. Samples of human cerebelli obtained from neurosurgical operations were examined for the presence of histamine-containing nerve fibers. In all samples, a moderately dense network of histamine-immunoreactive fibers was seen in the molecular layer. These fibers ran parallel to the Purkinje cell layer after traversing it perpendicularly. Numerous fibers were also seen in the granular cell layer. The results suggest that the human cerebellar cortex receives a direct input from histamine synthesizing hypothalamic neurons, as no other histamine-containing neurons have been found in human brain. PMID- 8247334 TI - Is the clustering of neurofibrillary tangles in Alzheimer's patients related to the cells of origin of specific cortico-cortical projections? AB - The spatial pattern of cellular neurofibrillary tangles (NFT) was studied in the supra- and infragranular layers of various cortical regions in cases of Alzheimer's disease (AD). The objective was to test the hypothesis that NFT formation was associated with the cells of origin of specific cortico-cortical projections. The novel feature of the study was that pattern analysis enabled the dimension and spacing of NFT clusters along the cortical ribbon to be estimated. In the majority of brain regions studied, NFT occurred in clusters of neurons which were regularly spaced along the cortical strip. This pattern is consistent with the predicted distribution of the cells of origin of specific cortico cortico projections. Mean NFT cluster size varied from 250 to > 12800 microns in different cortical tissues suggesting either variation in the size of the cell clusters or a dynamic process in the development of NFT in relation to these cell clusters. The formation of NFT in cell clusters which may give rise to the feed forward and feed-back cortico-cortical projections suggests a possible route of spread of NFT pathology in AD between cortical regions and from the cortex to subcortical areas. PMID- 8247335 TI - Compared effects of serotonin on the inspiratory activity of glossopharyngeal, vagal, hypoglossal and cervical motoneurons in neonatal rat brain stem-spinal cord preparations. AB - Experiments were performed on isolated brain stem-spinal cord preparations from neonatal rats, with a view to comparing the effects of serotonin (5-HT) on the rhythmic inspiratory activity recorded in the glossopharyngeal (IX), vagal (X), hypoglossal (XII) and cervical motor roots. Under normal medium, the inspiratory bursts recorded on the different roots presented slight changes in amplitude from one cycle to the other which were significantly correlated only between IX and X nerves. When the normal medium was replaced by a medium containing 5-HT (30 microM), the frequency of occurrence of the inspiratory bursts increased. After 6 min 5-HT superfusion, the inspiratory bursts observed in the IX, X and cervical recordings decreased slightly but significantly, whereas the XII inspiratory discharges decreased drastically. Upon returning to the control conditions, a total recovery was observed in all the recordings apart from the XII discharge, which remained depressed. Although 5-HT qualitatively depressed all the inspiratory motor outputs, the quantitatively drastic and long-lasting depression of the XII inspiratory activity was particularly noteworthy. 5-HT therefore seems to have specific effects on XII inspiratory activity which may be responsible for obstructive apnea in cases involving abnormal 5-HT biosynthesis mechanisms. PMID- 8247336 TI - Ciguatoxin extracted from poisonous moray eels Gymnothorax javanicus triggers acetylcholine release from Torpedo cholinergic synaptosomes via reversed Na(+) Ca2+ exchange. AB - Ciguatoxin (CTX) (0.1 pM to 10 nM) added to a suspension of Torpedo synaptosomes incubated in Ca(2+)-free medium caused no detectable acetylcholine (ACh) release. However, subsequent addition of Ca2+ caused a large ACh release that depended on time of exposure, dose of CTX and on [Ca2+]. Tetrodotoxin completely prevented CTX-induced Ca(2+)-dependent ACh release. Simultaneous blockade of Ca2+ channel subtypes by FTX, a toxin extracted from the venom of the spider Agelenopsis aperta, omega-conotoxin and Gd3+ did not prevent ACh release caused by CTX, upon addition of Ca2+. These results suggest that CTX activates the reversed operation of the Na+/Ca2+ exchange system allowing the entry of Ca2+ in exchange for Na+. It is concluded that Torpedo synaptosomes are endowed with Na+ channels sensitive to pico- to nanomolar concentrations of CTX. PMID- 8247337 TI - Extracellular potassium regulates the glutamine content of astrocytes: mediation by intracellular pH. AB - Based upon previous evidence that glutamine formation in astrocytes is pH sensitive and that raised extracellular K+ alkalinizes astrocytic cytoplasm, it was hypothesized that extracellular K+ might regulate glutamine formation. In this study, the free glutamine content of mouse cerebral astrocytes incubated with 0.1 mM glutamate and 0.1 mM ammonium increased by 80-90% when the extracellular K+ concentration was raised from 3 to 12 mM. The corresponding K(+) induced intracellular alkalinization of +0.13 pH units only partially reversed a glutamate-induced intracellular acidification of -0.24 pH units. By comparison, adjustment of extracellular pH from 7.4 to 7.8 shifted intracellular pH by +0.25 pH units, fully reversing the glutamate-induced acidification and increasing glutamine content by 120-180%. The effect of K+ on intracellular pH increased to +0.25 pH units in bicarbonate-buffered solution, suggesting that the regulation of glutamine formation by extracellular K+ is enhanced in the presence of bicarbonate. PMID- 8247338 TI - D1 and D2 dopamine receptors do not up-regulate in response to neonatal intrastriatal 6-hydroxydopamine lesions. AB - The extent of dopamine (DA) depletion appears to exert important influences on the plasticity of the DA system following lesions made in adulthood. The aim of this study was to determine if the extent of DA depletion has long-term effects on DA receptor regulation after early neonatal lesions. Early intrastriatal injections of 6-hydroxydopamine (6-OHDA) caused a dose-dependent loss of high affinity DA uptake sites and mu-opioid receptor patches evident in adulthood. DA receptors did not up-regulate in response to any degree of neonatal DA depletion. A patchy loss of D1 binding was evident following the neonatal lesions, although the loss was somewhat more severe and uniform with the highest dose of 6-OHDA (20 micrograms per striatum). There was also a slight decrease in D2 binding which was not dose-dependent. These results suggest that the consequences of early neonatal DA lesions are not dependent upon the degree of DA depletion, as the effects on DA receptor expression were similar regardless of the extent of the lesions. PMID- 8247339 TI - Saireito (a Chinese herbal drug)-stimulated secretion and synthesis of pituitary ACTH are mediated by hypothalamic corticotropin-releasing factor. AB - Administration of Saireito, a Saiko agent (a Chinese herbal drug), via a stomach cannula stimulates ACTH release and proopiomelanocortin, the precursor for ACTH, gene expression in the rat anterior pituitary. To study whether Saireito stimulated secretion and synthesis of ACTH are mediated by hypothalamic corticotropin-releasing factor (CRF), we examined the effect of passive immunization of endogenous CRF by i.v. administration of CRF antiserum on Saireito-increased plasma ACTH levels and proopiomelanocortin gene expression in the rat anterior pituitary, under pentobarbital anesthesia. CRF antiserum inhibited Saireito-induced plasma ACTH levels and proopiomelanocortin mRNA levels in the anterior pituitary. This result indicates that Saireito stimulates CRF neurons to increase CRF release, which stimulates secretion and synthesis of ACTH. PMID- 8247340 TI - Differential expression of c-fos and c-jun in two regions of the rat spinal cord following noxious colorectal distention. AB - Thoracolumbar and lumbosacral spinal segments that receive afferent input from the descending colon and rectum were stained immunocytochemically for c-Fos- and c-Jun-like proteins following repetitive, noxious colorectal distention (CRD). Noxious CRD (80 mmHg) resulted in significantly more c-Fos- and c-Jun-like immunoreactivity in the sacral dorsal horn than in the thoracic dorsal horn. In both regions of the spinal cord the increase in c-Fos-like immunoreactivity was at least twice that of c-Jun-like immunoreactivity. Basal levels of c-Jun but not c-Fos were observed in the thoracic intermediolateral nucleus (IML) and the sacral parasympathetic nucleus (SPN). Noxious CRD induced both c-Fos and c-Jun in the SPN, but not the IML. PMID- 8247341 TI - GABAA and GABAB receptors differentially regulate striatal acetylcholine release in vivo. AB - Microdialysis was used to study the effects of selective GABAergic agents on striatal acetylcholine (ACh) release in awake, freely moving rats. Local perfusion with the GABAA agonist muscimol dramatically reduced striatal ACh release, while the GABAB agonist baclofen caused only minor decreases in ACh release. Co-perfusion with the GABAA antagonist bicuculline diminished the muscimol-induced decrease in ACh release. Likewise, co-perfusion with the GABAB antagonist 2-hydroxysaclofen attenuated the baclofen-induced reduction in ACh release. Bicuculline alone markedly increased striatal ACh release, but 2 hydroxysaclofen by itself had no effect. These results suggest that GABA tonically regulates striatal ACh release primarily through stimulation of inhibitory GABAA receptors. PMID- 8247342 TI - Expression of cytokine receptors in cultured neuronal and glial cells. AB - We investigated mRNA expression of cytokine receptors in three different types of glial cells and two neuronal line cells by the RT-PCR method. Microglia expressed mRNA for receptors of IL-3, -4, -6, -7, GM-CSF, and M-CSF. Astrocytes were positive for receptors of IL-6, -7, GM-CSF, and M-CSF. Oligodendrocytes were positive for receptors of IL-3, -4, -7, GM-CSF, and M-CSF. Neuronal cells expressed receptors of IL-6 and GM-CSF with very low levels. This is the first demonstration of cytokine receptor mRNA expression in isolated glial and neuronal cells. PMID- 8247343 TI - Acute down-regulation of oxytocin and vasopressin mRNA levels following metrazole induced seizure in the rat. AB - Following our recent demonstration of metrazole-induced immediate-early gene expression in the hypothalamic supraoptic nucleus (SON), we have now performed a mRNA and transcription analysis to determine the consequences of metrazole treatment for neurohypophyseal peptide gene expression in male rats. Levels of hypothalamic vasopressin (VP) and oxytocin (OT) mRNA were significantly reduced at 2 and 4 h after metrazole (50 mg/kg, i.p.), whereas pro-dynorphin mRNA was significantly elevated at 2 h. No changes in mRNA levels were found at 8, 24 or 48 h after treatment. Another convulsant (kainic acid, 8 mg/kg, i.p.) elicited similar effects on VP and OT mRNAs at 2 h. Specific analysis of the SON, following metrazole, revealed an equivalent effect on VP and OT mRNA levels but a nuclear run-on assay did not detect any change in SON VP gene transcription at 0.5, 1 and 2 h after treatment. The results provide evidence of a novel mechanism which may provide an additional level of control in the regulation of neuropeptide gene expression. PMID- 8247344 TI - Beta-amyloid precursor protein (beta APP) as a marker for axonal injury after head injury. AB - It has been demonstrated recently that beta-amyloid protein (beta AP), generally associated with the plaques of Alzheimer's disease, can also be found in the brains of survivors of head injury. In this study the distribution of the beta AP precursor protein (beta APP) was examined immunohistochemically to determine if it is colocalized with beta AP in such cases. beta APP immunoreactivity was observed in neuronal perikarya in the neocortex and in dystrophic neurites surrounding beta AP immunoreactive plaques i.e. in a distribution similar to that seen in Alzheimer's disease. In addition, beta APP immunoreactivity was noted within white matter tracts where it marked damaged axons. However, no colocalisation of beta APP with beta AP was observed in any white matter region. These results indicate that processing of beta APP to produce beta AP occurs in the synaptic terminal field of axons and illustrate the utility of beta APP immunoreactivity as a general marker for axonal injury. PMID- 8247345 TI - Effects of GABAA receptor antagonism on inspiratory activities in kittens. AB - In anesthetized kittens (< 1 to 24 days old), the effects of GABAA receptor antagonism on phrenic, hypoglossal and cervical sympathetic discharges were examined by i.v. bicuculline infusions during hyperoxia and hypoxia. Administration of bicuculline during hyperoxia produced marked increases in the amplitudes of inspiratory nerve discharges. During hypoxic stimulation (10% O2), the amplitudes of inspiratory activities decreased towards or below those observed during hyperoxia; bicuculline reversed this depression and restored inspiratory discharges. Our results indicated that GABAA receptors were functional shortly after birth, acting to mediate influences shaping inspiratory activity during hyperoxic breathing and during conditions of increased chemical drive. PMID- 8247346 TI - Interhemispheric facilitation of the hand area of the human motor cortex. AB - We investigated interaction of the human bilateral hand motor area using two magnetic stimulators. A conditioning shock over the motor cortex facilitated the response evoked in the ipsilateral first dorsal interosseous muscle by a test shock over the contralateral motor cortex given under the limiting conditions of a small conditioning shock over a very small area and a small test shock. This effect was so weak that it was not elicited under slightly different conditions. We conclude that an excitatory, but weak, connection between strictly homotopic areas of the bilateral hand motor cortices exists in humans. PMID- 8247347 TI - A new metabotropic glutamate receptor agonist: developmental change of its sensitivity to receptors in the newborn rat spinal cord. AB - A novel metabotropic glutamate receptor agonist, (2S,1'R,2'R,3'R)-2-(2,3 dicarboxycyclopropyl)glycine (DCG-IV), reduced the monosynaptic excitation in newborn rat spinal cord rather than polysynaptic discharges at the nanomolar range without causing postsynaptic depolarization of motoneurones. Its inhibitory action on the monosynaptic excitation reduced in due course of time after birth. On the contrary, the inhibitory action of a metabotropic GABAB receptor agonist, baclofen, did not show marked developmental change. DCG-IV should be expected to have the potential to provide further useful information on the physiological function of metabotropic glutamate receptors. PMID- 8247348 TI - Effect of cyclosporine A on the morphine-induced place preference. AB - The effect of pretreatment with cyclosporine A, an immunosuppressant, on the morphine-induced place preference was examined in ddY and mu1 opioid receptor deficient CXBK mice. Morphine produced a significant preference for the drug associated place in ddY and CXBK mice. Administration of cyclosporine A alone to ddY and CXBK mice did not result in either preference or aversion for either the drug- or vehicle-associated place. On the other hand, pretreatment with cyclosporine A suppressed the place preference induced by morphine in ddY mice in a dose-dependent manner. However, in CXBK mice, pretreatment with cyclosporine A did not affect the morphine-induced place preference. These results suggest that cyclosporine A suppresses the reinforcing effect induced by morphine, and that this suppression by cyclosporine A may be mediated by mu1 opioid receptors. PMID- 8247349 TI - Cell responses to vertical and horizontal retinal disparities in the monkey visual cortex. AB - Because of the horizontal separation of both ocular globes, the projection angles are slightly different. These differences are commonly termed retinal disparities. Vertical and horizontal retinal disparities occur constantly in normal life. We have investigated the responses of single cells in cortical areas V1 and V2 of behaving Macaca mulatta monkeys to retinal disparities by using dynamic random dot stereograms. Our findings show that cortical visual cells are sensitive to both vertical and horizontal disparities. To calculate the distance between two objects in a three-dimensional space from horizontal disparities, it is necessary to know the fixation distance. It has been suggested that the horizontal gradient of vertical disparity contains information to estimate the fixation distance and therefore to scale horizontal disparities. We suggest that these cells sensitive to horizontal and vertical disparities represent a neural mechanism that provides information to the visual system in order to achieve a correct eye alignment and depth perception. PMID- 8247350 TI - In vivo administration of valproate reduces the nerve terminal (synaptosomal) activity of GABA aminotransferase in discrete brain areas of rats. AB - Previous in vitro experiments have indicated that neuronal GABA aminotransferase (GABA-T) differs from extraneuronal GABA-T in sensitivity to certain enzyme inhibitors, including the antiepileptic drug valproate (VPA). In the present experiments, it was studied if GABA-T activity in whole tissue and synaptosomes prepared from whole tissue of 11 brain regions of rats is affected in vivo by treatment with an anticonvulsant dosage of VPA. For this purpose, rats were treated with VPA, 200 mg/kg i.p., 15 min prior to dissection of brain regions. Synaptosomes were prepared from the 11 brain regions by a density gradient centrifugation method. In whole tissue samples of brain regions, no alteration of GABA-T activity was determined after VPA administration, except a small decrease in the tectum. In contrast, VPA significantly decreased GABA-T activity in synaptosomal fractions of substantia nigra, pons and medulla oblongata. These data substantiate that synaptosomal GABA-T is more susceptible to the inhibitory effect of VPA than the extra-synaptosomal enzyme. Furthermore, since VPA seems to be homogeneously distributed across brain regions, the observation of regional differences in inhibitory effects on GABA-T might indicate differences in the biochemical properties of GABA-T from different brain regions. PMID- 8247351 TI - Effects of extracellular ATP on hair cells isolated from the guinea-pig semicircular canals. AB - Externally applied ATP (100 microM) induced membrane currents in type I and type II vestibular hair cells enzymatically isolated from guinea-pig semicircular canals. In whole-cell voltage-clamp and with 140 mM K+ in the pipette solution, ATP evoked an inwardly directed current in 58% of the cells when held at potentials below -40 mV. In the remainder, external ATP produced an outward current. After block of the K currents, an inward current activated by ATP was revealed at -50 mV. Intracellular Ca2+ levels were monitored using the Ca2+ indicator Fura-2 and were found to rise in both hair cell types in response to ATP. These results strongly suggest that ATP directly controls the entry of Ca2+ into crista hair cells which can then further modulate K+ currents. PMID- 8247352 TI - Circannual rhythm in the effects of stress on the humoral immune response of the rat. AB - Among the seasonal immunological changes that have been observed are increase in spring-summer and decrease in autumn-winter humoral response, in unstressed subjects. In rats we show that, relative to controls, stress increased the antibody response to sheep red blood cells in July-August, and decreases it from September throughout February. We concluded, therefore, that stress reinforces the basic annual rhythm of this type of response. PMID- 8247353 TI - Cholecystokinin-A but not cholecystokinin-B receptor stimulation induces endogenous opioid-dependent antinociceptive effects in the hot plate test in mice. AB - The effects of intracerebroventricular administration of the cholecystokinin (CCK) analogue, BDNL, and the selective CCK-B agonist, BC 264, were determined using the hot plate test in mice. BDNL (0.2 nmol and 0.5 nmol) increased the jump and the paw lick latencies. These effects were blocked by the CCK-A antagonist MK 329 (0.02 mg/kg), supporting the involvement of CCK-A receptors in CCK-induced analgesia. In contrast, the selective CCK-B agonist BC 264 produced, at one dose (2.5 nmol), a slight decrease in the lick latency that was only antagonized by the CCK-B antagonist. Naloxone, but not naltrindole, antagonized BDNL-induced analgesia. The results suggest that activation of CCK-A receptors by BDNL leads to antinociceptive responses indirectly mediated by stimulation of mu-opioid receptors by endogenous enkephalins. PMID- 8247354 TI - Gonadotropin-releasing hormone agonist binding in tiger salamander nasal cavity. AB - Binding of the iodinated gonadotropin-releasing hormone (GnRH) agonist, buserelin, was examined in the nasal cavities of tiger salamanders using in vitro autoradiography. Binding of [125I]buserelin was seen within the chemosensory epithelium of the main nasal cavity and Jacobson's (vomeronasal) organ. Highest levels of binding were observed over the chemosensory neuron dendrites. Given the apparent lack of GnRH-immunoreactive fibers within the chemosensory epithelium as we have observed in a previous study, these observations suggest that GnRH may diffuse from fibers in the lamina propria of the chemosensory mucosa into the sensory epithelium to modulate chemosensory reception. PMID- 8247355 TI - Power spectral analysis of the electroencephalographic and hemodynamic correlates of propofol anesthesia in the rat: intravenous infusion. AB - Based on the tail-flick response to noxious thermal stimuli, we determined in the present study that effective antinociception could be achieved in adult male Sprague-Dawley rats 15 min after intravenous infusion of propofol at 60 mg/kg/h. Simultaneous power spectral analysis of the electroencephalographic (EEG) and systemic arterial pressure signals further revealed a concomitant depression of the activity of all EEG frequency bands (delta, theta, alpha, beta), alongside hypotension, negative inotropic and chronotropic actions, and attenuated baroreceptor reflex and vasomotor activity. These effects were congruent with a plasma concentration of propofol in the arterial blood of 1.70 +/- 0.13 micrograms/ml, as determined by high-performance liquid chromatography. PMID- 8247356 TI - The cerebrospinal fluid in patients with moyamoya disease (spontaneous occlusion of the circle of Willis) contains high level of basic fibroblast growth factor. AB - The level of basic fibroblast growth factor (bFGF) in cerebrospinal fluid (CF) was measured by an EIA in 15 patients with moyamoya disease, in 11 patients with atherosclerotic occlusive disease, and in 8 patients with spinal disc disease. In the moyamoya patients, bFGF was found in 13 out of 15 CF samples with a mean value of 101 pg/ml. However, bFGF was detected in only 4 out of 11 patients with atherosclerotic disease with a mean of 8 pg/ml. In all the patients with disc disease, bFGF was not detected. The elevated level of bFGF may play a crucial role in the pathogenesis of moyamoya disease. PMID- 8247357 TI - Inhibition of tryptophan hydroxylase by 6,7-dihydroxy-N-cyanomethyl-1,2,3,4 tetrahydroisoquinoline, a cyanomethyl derivative of dopamine formed from cigarette smoke. AB - 6,7-Dihydroxy-N-cyanomethyl-1,2,3,4-tetrahydroisoquinoline, a cyanomethyl derivative of dopamine formed from cigarette smoke, was found to inhibit the activity of tryptophan hydroxylase. The inhibition was non-competitive to the substrate L-tryptophan (the Ki value 7.25 +/- 0.81 microM), but not to the biopterin cofactor. The inhibition is irreversible. 6-Hydroxy-N-cyanomethyl tetrahydro-beta-carboline, a cyanomethyl derivative of serotonin, is inactive towards the hydroxylase. 6,7-Dihydroxy-N-cyanomethyl-1,2,3,4 tetrahydroisoquinoline may affect the serotonin biosynthesis in vivo as a consequence of cigarette smoking. PMID- 8247358 TI - Postnatal development of the anterior ethmoidal nerve in cats: unmyelinated and myelinated nerve fiber analysis. AB - This is the first quantitative electron microscopic study of anterior ethmoidal nerve in adult and newborn cats. The adult nerve comprises about 1,000 myelinated fibers including A delta (65%) and A beta (35%) fibers and 6,000 unmyelinated fibers. At birth, only 27% of the adult myelinated fibers complement is already present. The immaturity of the nerve is discussed in relation to that of the sneeze reflex. PMID- 8247359 TI - Parabigeminal, pretectal and hypothalamic afferents to rat's dorsal lateral geniculate nucleus. Comparison between albino and pigmented strains. AB - In adult pigmented and albino rats different fluorescent dyes were injected into the dorsal lateral geniculate nuclei of opposite sides. Differences between the strains occur mainly in parabigemino-geniculate and pretecto-geniculate projections. Both the major contralateral and the minor ipsilateral parabigemino geniculate projections in albinos were clearly smaller then those in pigmented rats. In pigmented rats but not in albinos the parabigemino-geniculate projections originated mainly from the region where the vertical meridian is represented and contained a small number of neurones projecting bilaterally. In each strain, a small number of retrogradely labelled neurones was found in the ipsilateral and contralateral lateral hypothalami. PMID- 8247360 TI - Brain-derived neurotrophic factor expression after long-term potentiation. AB - Long-term potentiation (LTP) of perforant-path dentate granule cell synapses, in awake rats, was followed by a time-dependent expression of brain-derived neurotrophic factor (BDNF) mRNA in dentate granule cells. This BDNF expression was blocked by the N-methyl-D-aspartate (NMDA) antagonist dizocilpine maleate (MK 801), which also blocked LTP induction, and by sodium pentobarbital, which shortens LTP persistence. These results suggest that BDNF may participate in the NMDA-receptor mediated cascade of events that result in LTP stabilization. PMID- 8247361 TI - Work smarter, not harder. PMID- 8247363 TI - The Clinton plan. PMID- 8247362 TI - My AIDS education Part I: Assuming the sick position. PMID- 8247364 TI - How to develop computer-assisted instruction programs. PMID- 8247365 TI - Computer assisted video instruction & community assessment. PMID- 8247366 TI - Worksite education for the RN-BSN student. PMID- 8247367 TI - Computerized adaptive testing: the future is upon us. PMID- 8247368 TI - Diploma programs in nursing accredited by the NLN 1993-94. PMID- 8247369 TI - The scholarship of application. PMID- 8247370 TI - My AIDS education. Part II: History lessons. PMID- 8247371 TI - Nurses' home health experience--Part I: The practice setting. PMID- 8247372 TI - How to plan financial support for nursing centers. PMID- 8247373 TI - Can nursing centers provide health care? PMID- 8247374 TI - A nurse managed center's client satisfaction survey. PMID- 8247375 TI - Models in community health care: findings from a national study of community nursing centers. PMID- 8247376 TI - My AIDS education. Part III: Political "realities". PMID- 8247377 TI - Health care reform proposals compared. PMID- 8247378 TI - NLN accreditation as a marriage of strangers. PMID- 8247380 TI - In search of excellence: an interview with Donna Peters. PMID- 8247379 TI - Self-evaluation in a complex system-wide school of nursing. PMID- 8247381 TI - Nursing education in the public's trust. A faculty of the community: no unreal loyalties for us. PMID- 8247382 TI - Nurses' home health experience--Part II: The unique demands of home visits. PMID- 8247383 TI - President's message. To prepare a preventionist. PMID- 8247384 TI - Living and dying peace and power: reflections on the last days of Charlene Eldridge. PMID- 8247385 TI - Functional alterations in dopamine systems assessed using drug discrimination procedures. AB - Numerous studies have suggested that Pb-induced perturbations of dopamine (DA) systems and DA functions could underlie the behavioral impairments attributed to Pb exposure. However, the precise nature of the effects of Pb on DA systems, either at the receptor/biochemical level or at the behavioral level have never been precisely delineated, much less interrelated. Substantial advances in the understanding of DA neuropharmacology provide new opportunities to more precisely elaborate Pb-induced changes in DA systems and DA function. Recently completed studies from our laboratories using drug discrimination procedures indicate that low-level postweaning Pb exposure produces a functional DA supersensitivity manifest at both the D1 and D2 receptor subtypes. Postnatal Pb exposure likewise induces functional DA supersensitivity, although this effect appeared to be restricted to the D2 receptor subtype. In conjunction with the drug discrimination studies, standard homogenate receptor binding assays of D1 and D2 receptors carried out in 5 different brain regions (striatum, nucleus accumbens, frontal cortex, midbrain and cerebellum) suggested a correspondence between D2 DA behavioral supersensitivity and Bmax changes found in nucleus accumbens, suggesting it as a possible site for Pb-induced supersensitivity to DA agonist stimulus properties. Collectively, the nature of the changes in DA sensitivity in the drug discrimination studies and the changes in D1 and D2 receptor number raise the possibility that Pb could, in part, produce a net functional autoreceptor agonism. PMID- 8247386 TI - Lead toxicity and alterations in opioid systems. AB - The opioid peptide system in the brain is probably the most extensive and diverse peptidergic transmission system. Three peptide precursors, pro-opiomelanocortin, proenkephalin and prodynorphin produce over 20 opioid peptides collectively known as the endorphins, enkephalins and dynorphins. Their effects are mediated by three receptors mu, delta and kappa, and the opioid system has control over several physiological functions including pain, locomotion, mood, diuresis, thermoregulation, stress, respiratory, gastrointestinal and cardiovascular function. Lead treatment (primarily using rat models) has shown that exposure to this metal in the perinatal period alters the development of endorphins and enkephalins, toxic effects which for the pro-opiomelanocortin products may be manifested at the gene level. Lead also alters the development of mu and delta receptors and biological responses to opioids such as analgesia, locomotion and stress responses. There are indications that the dynorphin/kappa opioid system is less affected than the mu and delta systems and this may suggest vulnerability to toxicity in the postnatal period as kappa systems are fully developed at birth whilst mu and delta systems are immature. In addition, hypothalamic and pituitary disruption of opioid peptides, plus alteration of stress-mediated activity by lead point to toxicity upon opioid controlled hormonal function. Comparative studies with other CNS neurochemicals and measures of blood lead levels suggest that opioid peptides are among the most sensitive neurotransmitter/neurohormonal systems to toxic insult by lead. PMID- 8247387 TI - Effect of perinatal lead treatment on morphine dependence in the adult rat. AB - The effect of perinatal lead exposure (at 300 and 1000 ppm in the maternal drinking water from conception to postnatal day 14) on the opioid withdrawal syndrome in adult offspring has been studied to assess if lead produces long term disruption of opioid systems manifested as altered morphine dependence. Dependence was induced in 50 day old rats by administration of morphine in osmotic mini-pumps implanted subcutaneously and delivering 5, 15 or 40 mg/kg/day. At postnatal day 55 an opioid withdrawal syndrome was precipitated by administration of naloxone (4 mg/kg i.p) and withdrawal behaviour scored over the next 30 min. Both objective (jumping, weight loss, weight of excreta, wet dog shakes, mouthing and face washing) and subjective (teeth chatter, ptosis, diarrhoea, irritability) measures were scored. 60 min after naloxone animals were killed and plasma corticosterone measured as a biochemical index of withdrawal. Morphine withdrawal scores and plasma corticosterone exhibited a clear dose relationship and there were no significant differences between 0 and 300 ppm lead exposed groups. However withdrawal scores in 1000 ppm lead-exposed animals were lower in 15 and 40 mg/kg morphine treated rats, predominantly associated with lower weight loss, wet dog shakes and mouthing responses. Paradoxically corticosterone levels were elevated in the 40 mg/kg morphine dose group. The results support other evidence that perinatal lead exposure can induce disruption in opioid functioning which persists to adulthood and suggest a possible link between lead and opioid addiction. PMID- 8247388 TI - Permeability of the blood-brain barrier to lead. AB - This review examines the kinetics and possible mechanisms of lead transport into brain across the microvessel endothelium (the blood-brain barrier). Although severe lead poisoning both in neonatal rats and in young children may cause microvessel damage, there is little evidence that there is either damage or even disturbance of specific transport mechanisms at blood leads < 80 micrograms/dl. When 203Pb was continuously infused intravenously into adult rats, radiotracer uptake into different brain regions was linear with time up to 4 hours, reaching spaces in relation to plasma of 6.6 - 8.2 ml/100 g in cerebral tissues at one hour. The concentration of free Pb+ in serum is of the order of 10(-12)M, the majority of lead being bound to protein and to sulfhydryl compounds, such as L cysteine. Transport into brain has been further studied during short vascular perfusion of one cerebral hemisphere of the rat with oxygenated and buffered physiological saline. This allows total control of the fluid perfusing the cerebral microvessels. In the absence of organic ligands for lead, 203Pb entered brain very fast, with a space of 9.7 ml/100 g in frontal cortex at one min. The presence of albumin, L-cysteine or EDTA abolished measurable uptake. Experiments designed to reveal a role for the anion exchanger or calcium channels gave negative results. However, the effects of potassium depolarization and of varying pH indicated that the lead species passively entering the endothelium might be PbOH+. Experiments with various metabolic inhibitors, including vanadate, suggested that Pb uptake in the endothelium is mitigated by active back transport of lead into blood by the Ca-ATPase pump.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247389 TI - Electrophysiology of lead intoxication: effects on voltage-sensitive ion channels. AB - Neuronal function depends on the activity of a variety of voltage-sensitive, ion specific membrane channels, including channels permeable chiefly to sodium, potassium, and calcium. The plasma membranes of many neurons contain several types of each class of channel. In general, heavy metal ions exert little effect on voltage-sensitive sodium or potassium channels, but inhibit ion flow through voltage-sensitive calcium channels (VSCC). The literature abounds with descriptions of different types of calcium channels in vertebrate neurons. These descriptions suggest that there are many physiologically and pharmacologically distinct calcium channels, some of them possibly cell-type specific. Among the heavy metals, Pb2+ is one of the most potent inhibitors of VSCC in both vertebrate and invertebrate neurons. Some heavy metals, including Ni2+ and Cd2+, are fairly selective against certain types of calcium channels. Limited evidence suggests that Pb2+ inhibits all calcium channel types within a given cell, with only minor differences in potency. However, there appear to be substantial differences among cell types in the concentration dependence of calcium channel inhibition by Pb2+. Therefore, to appreciate the range of effects of Pb2+ on calcium channels throughout the nervous system, it will be important to examine a large number of cell types. Pb2+ is highly permeable through at least some types of VSCC. Entry of Pb2+ into the neuronal cytoplasm through VSCC, followed by disturbance of intracellular functions, may be a major mechanism of Pb2+ neurotoxicity. PMID- 8247390 TI - Current issues in human lead exposure and regulation of lead. AB - Concern about lead as a significant public health problem has increased as epidemiological and experimental evidence has mounted regarding adverse health effects at successively lower levels of lead exposure. This concern has led to downward revision of criteria for acceptable blood lead concentrations to the 10 micrograms/dL mark now designated by EPA as a target level for regulatory development and enforcement/clean-up purposes. Much progress has been made in reducing lead exposures during the past 10-15 years, with marked declines evident both in air lead and blood lead concentrations in parallel to the phase-down of lead in gasoline and notable decreases in food lead exposure due to elimination of lead soldered cans by U.S. food processors. With the lessening of exposure from these sources, the importance of other components of multimedia exposure pathways has grown and stimulated increasing regulatory attention and abatement efforts to reduce health risks associated with lead exposure from drinking water, from lead-based paint, and from household dust and soil contaminated by deteriorating paint, smelter emissions, or various other sources. Increasing attention is also being accorded to reduction of occupational lead exposures (including those related to lead abatement activities), with particular concern for protection of men and women during their reproductive years. PMID- 8247391 TI - Lead-induced changes in learning: evidence for behavioral mechanisms from experimental animal studies. AB - Lead is probably the most studied of the neurotoxic agents, both in humans and in animal models. Research has focused on learning impairment and other behavioral consequences produced by developmental exposure. In children, lead exposure results in deficits in such global measures as IQ, as well as more specific deficits that are suggestive of attentional deficit disorder. Research in animals has also clearly demonstrated learning and memory deficits as a consequence of developmental lead exposure. Experiments performed in monkeys implicate the same behavioral problems as those observed in children: increased distractibility, inability to inhibit inappropriate responding, and perseveration in behaviors that are no longer appropriate. For example, lead-treated monkeys were impaired in their ability to perform discrimination reversal task, but not on the initial visual discrimination task. Deficits were more severe in the presence of distracting irrelevant stimuli. Lead-treated monkeys displayed severe perseveration on one button on a task which required them to alternate responding between two buttons. Lead-treated monkeys displayed memory impairment on a task requiring them to remember a previously observed stimulus or position, which was at least in part the result of interference from responses from previous trials. Lead-treated monkeys exhibited a higher rate of response on an intermittent schedule of reinforcement, and had difficulty inhibiting responding when required. Thus, for many of the tasks on which monkeys have been found to display learning and/or memory impairment, the deficit can be attributed at least in part to an attentional deficit and/or perseverative behavior. PMID- 8247392 TI - Functional implications of postural disequilibrium due to lead exposure. AB - Measurement of postural equilibrium has been employed as an indirect indicator of functional status of the nervous systems of 109 children (mean age: 5.8 +/- 0.78 years) from the Cincinnati Lead Program Project. The geometric mean blood lead for the first five years of life (PbB05) was 11.9 +/- 1.5 micrograms/dL. Postural sway associated with upright balance was noninvasively quantitated with a microprocessor-based force platform and four tasks performed for 30 sec each by the subjects. A covariate-adjusted multiple regression analysis showed statistically significant associations between PbB05 and the postural sway for the task requiring primarily vestibular and/or proprioceptive systems input, implying potential functional impairment. These findings raised several new issues which are addressed here: (1) A dynamic task has been developed to further test the effect of perturbing those afferents' functional capabilities. (2) A method has been developed to quantitate the stability boundary of each subject to better characterize the limits of functionally-safe postural sway. (3) There is a need to perform the postural sway in a shorter duration than 30 sec so that children younger than five years of age can be tested for early identification of Pb-induced functional impairment of postural equilibrium. PMID- 8247393 TI - Auditory and visual dysfunction following lead exposure. AB - The effects of lead exposure on cognitive function have been intensively studied during the past decade, but relatively little effort has been made to understand the impact on sensory function. Subtle impairments of visual and/or auditory processing, however, could have profound effects on learning. The objectives of this paper are to review what is known about the effects of lead exposure on visual and auditory function and to identify related research needs. In particular, the effects of lead exposure on sensory function in children, which have not been studied adequately, will be discussed. Evidence from human and animal studies reveal that lead exposure impairs auditory function. The cochlear nerve and more central structures appear to be preferentially sensitive in both developing and mature humans and experimental animals. Elevations in hearing thresholds and increased latencies of brainstem auditory evoked potential have been reported at low-moderate levels of lead exposure. Higher doses of lead increase the threshold of the auditory nerve action potential, produce segmental demyelination and axonal degeneration of the cochlear nerve, but appear to have no effect on cochlear microphonics or structure. Lead exposure affects both the retina and visual cortex of the developing and mature visual system. Low to moderate level developmental lead exposure produces selective rod deficits which can be detected with electrophysiological and behavioral techniques. At slightly higher levels of lead exposure the visual cortex is affected. A wide range of functional and neurochemical effects on retinal function occurring at blood lead levels below 20 micrograms/dl, the current level of concern, have been observed in rats. Structural, biophysical and photochemical similarities of rods in rats, monkeys and humans argue the relevance of this data for pediatric lead screening. To date, however, rod-mediated visual functions have not been examined in lead exposed children. Undetected sensory deficits of these kinds may have profound impact on the motor and mental development of children as well as on the quality of life of affected adults. There is clearly a need for more extensive sensory testing in children and workers to screen for lead-induced health effects and in animal models to clarify the mechanisms of lead neurotoxicity. PMID- 8247394 TI - Trends in the management of childhood lead poisonings. AB - It is unknown whether prompt medical management (with or without chelation therapy) and environmental intervention have beneficial effects other than stopping the progression towards symptomatic childhood lead poisoning. Stated differently, does prompt intervention have substantive beneficial effects or are untreated lead toxic children with blood lead values between 25-54 micrograms/dl irrevocably damaged by the time of their identification. We are carrying out a prospective treatment outcome study with CaNa2EDTA (when indicated) at our Center to hopefully answer this critical question, within the context of a multidisciplinary study. The results in 162 children indicate that environmental and medical management produce significant reductions in blood lead, erythrocyte protoporphyrin and the lead diuresis during a CaNa2EDTA provocative test. However, CaNa2EDTA treatment failed to decrease bone lead values dramatically, measured by L-line x-ray fluorescence, six months after enrollment in any patient group (treated or untreated with CaNa2EDTA). The uses of L-line x-ray fluorescence in this study and K-line x-ray fluorescence measurements of lead in bone in other reported studies open a wide time window of months to years of lead exposure, compared to 30-45 days, the time of exposure captured by blood lead levels. As with all chelating agents, DMSA should be administered to children in lead free housing, after this drug's toxicity is more widely assessed. The potential capability of DMSA to ameliorate neurobehavioral deficits produced by lead must be systematically assessed and compared with CaNa2EDTA in a randomized, controlled study before the use (s) of either drug become uncritically accepted as the treatment of choice for childhood lead poisoning, in addition to full abatement. PMID- 8247395 TI - Conceptual and practical advances in the measurement and clinical management of lead toxicity. AB - The purpose of this paper is to discuss issues which relate to the risks and benefits of pharmacologic intervention in children with elevated blood lead (BPb) concentrations. In January, 1991, 2,3-dimercaptosuccinic acid (DMSA; succimer or CHEMET) was approved by the FDA for administration to children with blood lead concentrations of 45 micrograms/dl or higher. Thus, it is now considerably simpler to treat children with elevated BPb's. DMSA is orally actively, relatively safe and specific, and can, in some situations, be used on an out patient basis. The author is concerned that the recent CDC statement on lead poisoning, whose aim is to reduce (by various means) children's BPb's below 10 micrograms/dl, may unintentionally encourage pediatricians to treat children with relatively low BPb's with DMSA, both for medical and legal reasons. Should children with BPb's of less than 45 ug/dl be treated, and why? The risks and benefits of treatment in such children are uncertain. A randomized, placebo controlled, double-blind drug intervention trial is needed (and justified) in order to more carefully assess the benefits and the risks associated with the treatment of lower BPb's. PMID- 8247396 TI - Lead in bone: storage site, exposure source, and target organ. AB - The primary site of lead storage is in bone but relatively little attention has focused on this physiological compartment. Recent advances in measurement technology now permit the direct in vivo quantitative measurement of lead in bone, and this measure has great use in clinical and epidemiologic studies. Lead in bone is not a physiological sink, but can be mobilized back into the circulation in response to normal or pathological changes in mineral metabolism. Bone lead may be a significant source of target organ exposure under certain conditions, such as pregnancy, kidney disease, and menopause. Finally, the accumulation of lead in bone cells may have toxic consequences for bone status, and some of the mechanisms by which lead could affect bone mineral metabolism may also play a role in other target organ effects of lead. PMID- 8247397 TI - Beyond LOEL's, p values, and vote counting: methods for looking at the shapes and strengths of associations. AB - Determining the shape of dose-response relationships, including the possible existence of thresholds, is a central concern in the epidemiology of neurotoxins. Assessing the strengths of associations within a paper, and for the literature as a whole, are also critical concerns. Modern statistical and epidemiologic methods have considerable advantages for answering these questions. This is illustrated by using non-parametric smoothing to look for the existence of thresholds or non linearities in the neurotoxic effects of lead, the use of multiple analyses and robust methods to test the stability of the association between dentine lead and children's IQ in the study of Needleman et al., 1979, and the examination of the stability of the association between full scale IQ and blood lead across studies. Non-parametric smoothing shows the dose response relationship between McCarthy score and blood lead reported by Bellinger et al., (1991) has no threshold down to blood lead levels of 1 microgram/dl. The association between blood lead and hearing threshold reported by Schwartz and Otto (1991) also has no threshold down to the lowest concentrations seen in that study. The association between dentine lead and IQ in the Needleman et al. (1979) data holds in the full data set with no exclusions and with control for age, is insensitive to exclusions of subjects or inclusions of other covariates, and to the use of robust estimation techniques. Nonparametric smoothing shows the association is supported across the entire range of lead exposure and is not driven by a few outliers. Examination of effect size estimates from the IQ-blood lead studies shows moderately good concordance. This suggests a causal relationship. A meta-analysis of those studies indicates that an increase in blood lead from 10 micrograms/dl to 20 micrograms/dl is associated with a reduction of 2.5 points in full scale IQ. The effect was highly significant. PMID- 8247398 TI - Blockade of mammalian and invertebrate calcium channels by lead. AB - We have compared the effects of Pb2+ on voltage-dependent calcium channels of the marine mollusc Aplysia, studied with a two electrode voltage clamp, with those on calcium channels in cultured rat dorsal root ganglion (DRG) neurons studied with whole cell patch clamp. In both preparations Pb2+ was a potent blocker of calcium channel currents at concentrations that did not significantly affect potassium and sodium currents. The blockade was concentration dependent and the percentage of blockade was reduced when the concentration of the charge carrier was elevated. In Aplysia the threshold Pb2+ concentration was about 1 microM, and the Hill coefficient near 1.0 under all conditions. Pb2+ did not significantly change inactivation but shifted the voltage dependence of activation to hyperpolarized voltages in a dose-dependent manner. The blockade of calcium currents by Pb2+ was highly voltage dependent and increased with depolarization. Rat dorsal root ganglion cells exhibit three different types of voltage-dependent calcium channels (N, L and T) which can be distinguished by the potential at which the channel activates or inactivates and by their sensitivity to pharmacologic antagonists. The IC50 for blockade of the L current was 1.03 microM with a Hill slope of 1.15. Currents elicited by voltage steps which activate N plus L currents had an IC50 of 0.64 microM and a Hill slope of 1.16. T currents were less sensitive, having an IC50 of 6 microM. Sodium and potassium currents were relatively unaffected in both preparations at concentrations at which the calcium channel was blocked more than 60% (1 microM or 200 microM respectively). The blockade in DRG neurons was less voltage-dependent and reversible than that in the invertebrate model system. These observations indicate that Pb2+ is a potent, reversible and selective blocker of voltage-dependent calcium channels at low concentrations. PMID- 8247399 TI - The effects of inorganic lead on voltage-sensitive calcium channels differ among cell types and among channel subtypes. AB - The whole-cell version of patch clamping was used to compare the effects of acute in vitro exposure to inorganic lead (Pb2+) on voltage-sensitive calcium channels in cultured N1E-115 mouse neuroblastoma cells and E18 rat hippocampal neurons. Free Pb2+ concentrations in salines with a high lead-buffering capacity were measured with a calibrated Pb(2+)-selective electrode. Previously, we found that N1E-115 neurons contain low voltage activated, rapidly inactivating (T) channels and high voltage activated, slowly inactivating (L) channels. Pb2+ inhibits both channel subtypes in N1E-115 cells, with some selectivity against L-type channels (IC50 approximately 700 nM free Pb2+ for L-type channels, 1300 nM free Pb2+ for T type channels; Audesirk and Audesirk, 1991). In addition to T-type and L-type channels, cultured E18 rat hippocampal neurons have been reported to contain high voltage-activated, rapidly inactivating (N) channels. In our experiments with 5 to 20 day old cultures, almost all neurons showed substantial L-type current, approximately half showed significant N-type current, and fewer than 5% showed significant T-type current. We found that Pb2+ is somewhat selective against L type channels (IC50 approximately 30 nM free Pb2+ in 10 mM Ba2+ as the charge carrier, 55 nM in 50 mM Ba2+) compared to N-channels (IC50 approximately 80 nM free Pb2+ in 10 mM Ba2+, 200 nM in 50 mM Ba2+). These results suggest that the effects of Pb2+ on calcium channels of vertebrate neurons vary both among cell types and among channel subtypes. PMID- 8247400 TI - Low-level lead exposure in cultured astroglia: identification of cellular targets with vital fluorescent probes. AB - The effects of lead (0.0, 0.1, or 1 microM) on subcellular sites in primary astroglial cultures were quantitated with the use of vital fluorescent probes (fluorescence bioassays). Evaluation of cellular glutathione levels with monochlorobimane revealed a reduction in glutathione content after only 7 hr of Pb treatment to 77 and 82% of control values for 0.1 and 1.0 microM Pb, respectively. A further decrease in intracellular glutathione levels (to 74 and 56% of control values, respectively) was observed after 24 hr. Glutathione content returned to control levels by 48 hr, and exceeded control levels after 6 days (122 and 159% of control values) and 9 days (135 and 154% of control values) of lead treatment. Whereas glutathione has been shown by others to protect target organs from metal toxicity, these findings suggest a compensatory response by astroglia to the effects of Pb. Alterations in astroglial mitochondrial membrane potential were measured with the use of rhodamine 123. The membrane potential dependent partitioning of rhodamine 123 was reduced following 14 days of Pb exposure (0.1 or 1.0 microM) to 75% of control value, indicating that Pb may act to dissipate the electrochemical gradient in astroglial mitochondria. Carboxyfluorescein diacetate was used to evaluate gap junctional intercellular communication (GJIC) between astroglia by fluorescence recovery after photobleaching. No difference between control and low-level Pb treated astroglia was found. Our results indicate that a Pb-stimulated increase in astroglial glutathione occurs after a lag period during which levels are decreased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247401 TI - Lead suppresses chimeric human transferrin gene expression in transgenic mouse liver. AB - The major iron-transport protein in serum is transferrin (TF) which also has the capacity to transport other metals. This report presents evidence that synthesis of human TF can be regulated by the metal lead. Transgenic mice carrying chimeric human TF-chloramphenicol acetyl transferase (CAT) genes received lead or sodium salts by intraperitoneal injections or in drinking water. Transgene expression in liver was suppressed 31 to 50% by the lead treatment. Lead regulates human TF transgenes at the mRNA level since liver CAT enzyme activity, CAT protein, and TF CAT mRNA levels were all suppressed. The dosages of lead did not alter synthesis of the other liver proteins, mouse TF and albumin, as measured by Northern blot analysis of total liver RNA and rocket immunoelectrophoresis of mouse sera. Moderate levels of lead exposure were sufficient to evoke the human TF transgene response; blood lead levels in mice that received lead acetate in drinking water ranged from 30 micrograms/dl to 56 micrograms/dl. In addition to suppressing expression of TF-CAT genes in transgenic mice, lead also suppressed synthesis of TF protein in cultured human hepatoma HepG2 cells. The regulation of human TF apparently differs from the regulation of mouse TF which is unresponsive to lead exposure. PMID- 8247402 TI - Solubility of lead as lead (II) chloride in HEPES-Ringer and artificial seawater (Ca-ASW) solutions. AB - Total dissolved Pb was measured in a number of commonly used physiological solutions by electrothermal atomization atomic absorption spectrometry (ETA-AAS). In HEPES-buffered solutions (pH 7.30) the concentration of total Pb in solution ("measured" Pb) was only 77% of nominal Pb up to 20 microM added Pb, where experiments were undertaken at room temperature (22 +/- 1 degree C). However, test solutions equilibrated at 37 +/- 1 degree C contained 99% of added Pb up to 2 microM. Above this nominal concentration, percentage recoveries dropped to approximately 72% at a nominal concentration of 20 microM. Tris-buffered artificial seawater (CaASW) solutions (pH 7.60) contained more dissolved Pb compared to HEPES-buffered solutions at 22 degrees C. However, increasing the calcium concentration in ASW appeared to increase precipitation of PbCl2. Concentration-corrected dose-response relationships were plotted from previously published data on the effects of Pb on voltage-activated calcium channels of rat dorsal root ganglion (DRG) cells and Aplysia neurons. The plots suggest that the inhibitory effects of Pb on rat DRG cells may prevail at concentrations of Pb even lower than reported previously when measurements were made at 22 degrees C. However, increasing the temperature to 37 degrees C resulted in closer agreement between measured and nominal dose-response curves. The measured dose-response curves for the Tris-buffered ASW solutions closely followed those of the nominal up to 200 microM Pb. In ASW solutions containing 40 mM calcium, PbCl2 was precipitated at Pb concentrations greater than 200 microM. PMID- 8247403 TI - New directions in the toxicokinetics of human lead exposure. AB - An important determinant of body lead (Pb) burden and Pb toxicity in exposed humans is Pb metabolism, or more correctly, Pb toxicokinetics. It affects the former through the quantitative processes of uptake, distribution and retention/excretion and the latter via delivery of toxic doses to cellular/molecular sites of action. Pb toxicokinetics has useful application in understanding Pb's behavior in populations. Several of these applications have been studied and results are presented for the toxicokinetic basis of dose neurotoxic effect relationships in selected longitudinal studies and the use of toxicokinetic modeling for estimation of body lead burden in early populations. Three well-known, ongoing longitudinal studies of developmental neurotoxicity--in Boston, Cincinnati, and Port Pirie, Australia--involve cohorts who differ markedly as to their pre- and postnatal lead exposure profiles. Toxicokinetic examination of these exposure differences helps to explain the temporal variability seen in blood Pb-toxic effect relationships and supports a causal role for lead. Toxicokinetic models of Pb uptake and in-vivo behavior are increasingly being considered for estimating Pb-B levels in lieu of direct measurement. A linear biokinetic model, using reliable input data for natural/prehistoric levels of Pb in sources, was applied to estimation of prehistoric/preindustrial children's blood lead. A range of 0.06 to 0.12 microgram/dl was estimated for two lead intakes. These estimates are still two orders of magnitude (85 to 165-fold) lower than the newly issued CDC toxicity guideline for children of 10 micrograms/dl. Lastly, the toxicokinetics of lead in bone, particularly its resorption with metabolic stimuli, is of concern, particularly for "baby boom" women who are either of childbearing age or approaching menopause and who had greatly elevated environmental lead exposures in the 1940s to 1970s. PMID- 8247404 TI - New mechanisms in lead biodynamics at ultra-low levels. AB - It has been found that the mean concentration of 15 micrograms Pb/g bone ash in typical Americans today is 1000 times greater than the natural level of lead in Homo sapiens sapiens determined from analyses of ancient human bones. Since body burdens of lead only 3 to 5-fold greater than typical induce overtly poisonous effects, unrecognized dysfunctions caused by lead are probably widespread among Americans. Previous toxicological studies by others to delineate ill effects were made by comparisons of biological systems to which lead was added with systems to which no lead was added. It is now seen that such studies dealt entirely with unnatural mechanisms in lead biodynamics, and provided no knowledge of the operation of unperturbed natural mechanisms, because "non-leaded" controls were actually highly traumatized by unrecognized excessive lead exposures. New work should be carried out that would use an ultra-low lead diet to grow rats possessing "natural" lead levels which could be used as controls. Metabolisms of lead in these controls should be compared with metabolisms of lead in biological systems of rats traumatized by excessive lead exposures corresponding to typical levels within people today. Such comparisons would disclose previously unknown characteristics of intra-cellular processing of lead within osseous, neuronal, hepatic, renal and muscle tissues, and in red blood cells and sera, together with sizes and residence times of lead reservoirs in those tissues during fetal growth, maturation and adult aging in rats exposed to natural lead levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247405 TI - Lead, reproduction and development. AB - Lead remains a significant occupational health and public health problem. Excessive lead exposure, not only to men exposed at work, but also from lead based paint in older housing and from soil, still persists in the community. From the perspective of human reproduction, lead is known to cause a number of adverse outcomes in both men and women. Reported effects in men include reduced libido, effects on spermatogenesis (reduced motility and numbers, increased normal morphology), chromosomal damage, infertility, abnormal prostatic function and changes in serum testosterone. Most of these effects have been seen in lead exposed workers, and some are at levels presently considered 'acceptable'. Effects in women include infertility, miscarriage, premature membrane rupture, pre-eclampsia, pregnancy hypertension and premature delivery. Again, recent research findings indicate that lead may be toxic at levels previously thought to have no effect. The evidence implicating a possible teratogenic effect of lead is much less clear. While there is some indication of an association of lead with the production of anomalies, any dose-response relationship or causal mechanism has yet to be established. Therefore, while lead does affect human reproduction, the question: Is lead a teratogen? is yet to be answered. PMID- 8247406 TI - Effects of inorganic lead on the differentiation and growth of cortical neurons in culture. AB - Lead exposure has devastating effects on the developing nervous system, producing morphological, cognitive, and behavioral deficits. To elucidate some of the mechanisms of lead neurotoxicity, we have examined its effects on the differentiation of several types of cultured neurons. Previously, we reported the effects of inorganic lead on several parameters of growth and differentiation of E18 rat hippocampal neurons and two types of neuroblastoma cells cultured in medium with 2% fetal calf serum (FCS) (Audesirk et al., 1991). In the present study, we report the effects of concentrations of lead ranging from 1nM to 1 mM on the differentiation of hippocampal neurons cultured in medium containing 10% FCS. In addition, we investigated lead effects on neurons isolated from the motor cortex region of the E18 rat embryo. Cortical neurons were exposed to lead in concentrations ranging from 0.1 nM to 1 mM in medium with either 10% FCS or 2% FCS for 48 hr. The effects of lead tended to be multimodal. Neurite initiation, which is highly sensitive to neurotoxic compounds, was inhibited by lead at both high and low concentrations, with no effects at intermediate levels. Medium with 10% FCS enhanced certain growth parameters and tended to reduce the effects of lead. There was an overall consistency in the effects of lead on motor cortex and hippocampal neurons. PMID- 8247407 TI - The effects of lead exposure on learning in a multiple repeated acquisition and performance schedule. AB - This study sought to determine the selectivity of Pb-induced changes in learning, as distinct from non-specific or performance effects, and to explore the nature of the underlying error patterns contributing to any learning deficits. To accomplish this, rats were chronically exposed to 0, 50, or 250 ppm Pb acetate in drinking water from weaning and trained on a multiple repeated acquisition (RA) and performance (P) schedule beginning at 55 days of age. The RA component required the rat to learn a new 3-member sequence of responses during each experimental session (Center Right Left, RLC, CLR, RCL, and LRC), while the correct sequence of responses for the P component was constant across sessions (LCR). Significant decrements in accuracy on the RA component but not on the P component were found in Pb-exposed groups compared to control, effects that could not be attributed to differential rates of responding. Analyses of error patterns revealed that the effects of Pb exposure on RA accuracy levels derived from two sources. The first consisted of a perseveration of P-like sequence responding (LCR) even during the RA component. Secondly, Pb exposure increased perseverative responding on a single lever, even though the schedule itself never directly reinforced such repetitive responding. The increase in frequency of these two types of perseverative behavior was incompatible with acquisition of non P-like sequences during the RA component. Adding a 5 sec tone to the light stimuli signalling the transition between RA and P components of the multiple schedule failed to attenuate these effects of Pb, suggesting that deficits in stimulus control were not the sole behavioral mechanism of these impairments. Examination of individual data revealed the presence of both 'learners' and 'non-learners' in each group, with the prevalence of the latter being suggestively higher in Pb exposed groups than in controls. These findings may be relevant to the classroom setting, where periods requiring learning may frequently be interspersed with periods of performance of learned skills. PMID- 8247408 TI - Exposure to sublethal levels of waterborne lead alters reproductive behavior patterns in fathead minnows (Pimephales promelas). AB - Lead (Pb) caused multiple effects on reproductive behavior and overall reproductive success. Adult fathead minnows (Pimephales promelas) were acclimated at a 16L:8D photoperiod to stimulate reproductive development. Reproductively mature adults were separated as male-female pairs and maintained for 4 weeks in either 0.0 or 0.5 ppm Pb. High lead accumulations occurred in testes and ovaries of treated fish; lead concentrations in control fish gonads were not detectable. Lead suppressed spermatocyte production and retarded ovarian development, although no lead-induced gonadosomatic index changes for either sex were noted. Lead decreased the number of eggs oviposited, increased interspawn periods and suppressed embryo development. Control males displayed maximum secondary sex characteristic development (banding, tubercle formation, head and eye darkening); lead-exposed fish displayed less. Control males spent more time in ceiling directed behaviors associated with nest preparation and maintenance than lead exposed. These variables were affected differentially with respect to stage of reproductive maturity at time of lead exposure, i.e., fish displaying greater secondary sex characteristic development before exposure were less affected by lead than those fish that showed less development. PMID- 8247409 TI - The speciation of lead in erythrocytes in relation to lead toxicity: case studies of two lead-exposed workers. AB - Lead toxicity is known to be subject to individual susceptibility. This study compares two lead-exposed subjects, one (A; blood Pb 1800 micrograms/L) who remained totally asymptomatic, the other (B; blood Pb 1610 micrograms/L) who showed symptoms of toxicity. We have assessed the speciation of lead in the intra erythrocyte proteins in these patients and have examined its significance in relation to clinical toxicity. Chromatographic separations of erythrocyte haemolysates from these patients showed a metallothionein-like lead containing protein. It was demonstrated that in patient A, most (approximately 70%) of the erythrocyte lead was associated with this protein, whilst in patient B the protein only contained about 20% of the total lead, with significant amounts bound to high molecular weight proteins, including Hb. Further purification of this protein from each patient showed it to contain a number of constituents, one in particular being the major lead-binding species. This component was more abundant in patient A and, relative to patient B, contained a higher proportion of lead. These results suggest that this protein may act to sequester lead into a non-bioavailable form, hence protecting the body from lead toxicity as with patient A. PMID- 8247410 TI - New dimensions of lead neurotoxicity: redefining mechanisms and effects. PMID- 8247411 TI - Lead-protein interactions as a basis for lead toxicity. AB - The interaction of lead (Pb) with proteins may represent a fundamental mechanism by which Pb exerts toxicity. In this overview, various factors which influence the interaction of Pb with proteins will be discussed. Pb interacts with enzyme functional groups, and high-affinity metal-binding proteins, such as Pb-binding proteins and metallothioneins, can mediate this Pb-enzyme interaction. Many other factors influence Pb-protein interactions including ligand competition and binding affinities; protein folding and the nature of the metal-binding site; rates of protein synthesis and degradation; and intracellular localization of the ligand and metal. The remainder of this overview will focus on specific examples of important proteins known to be influenced by Pb or which hypothetically may be influenced by Pb. Gaps in knowledge and important research needs are emphasized. Many of the factors discussed play a role in the relative sensitivity of various enzymes in heme biosynthesis to Pb. Disruption of this critical pathway by Pb may result in neuropathologies and accumulation of neurotoxic heme precursors. High affinity metal-binding proteins have been shown to play a role in mediating Pb inhibition of the octameric Zn-containing enzyme, ALA dehydratase. Knowledge of regional localization in brain and the postnatal ontogeny of the high-affinity metal-binding proteins may be pivotal in understanding Pb neurotoxicity. Other specific examples related to or potentially related to Pb toxicity which are discussed include nucleic acid binding proteins, calmodulin, protein kinase C, and carbonic anhydrase. These proteins will serve as models to understand some basic principles and differences in Pb-protein interactions. PMID- 8247412 TI - A nuclear matrix protein stabilized by lead exposure: current knowledge and future prospects. AB - p32/6.3, a low-abundance, highly conserved nuclear protein, is a target for lead. Very few low abundance nuclear proteins have been described and no others have been associated with lead. Its wide distribution and conservation indicate a fundamental nuclear role. Further, it increases many fold in grey matter of brain and spinal cord during the neonatal period; there are no other identified nuclear proteins which serve as markers for this period of nervous system development. There are several links between lead and p32/6.3. It is a major component of lead induced intranuclear inclusion bodies from the kidney. Its accumulation in kidney is a relatively early event in the process of lead intoxication. Exposure to lead increases p32/6.3 in mouse neuroblastoma 2a cells within one day, blocking its degradation almost completely. These observations suggest that lead either structurally alters p32/6.3 or inhibits a protease for which p32/6.3 is a substrate. In these lead-treated cells nuclear envelope invaginations and small nuclear bodies increase. The possible involvement of lead and p32/6.3 with the formation and movement of nuclear bodies is discussed. PMID- 8247413 TI - Neural cell adhesion molecules, neuronal development and lead toxicity. AB - The major families of neural cell recognition molecules and the mechanisms by which they regulate neural development are described. The role of the neural cell adhesion molecule (NCAM) isoforms in organizing tissue structure and pattern is discussed in detail and their degree of sialylation is proposed to regulate the intimacy of cell-cell interaction, including that of synapse formation. The ability of chronic low level lead to impair normal NCAM sialylation pattern during postnatal development is discussed and this is suggested to perturb synapse selection procedures in a manner which may account for subsequent learning deficits in the adult rodent. In support of this proposal, change in hippocampal NCAM sialylation is demonstrated to occur in the synapse-specific isoform during the acquisition and consolidation of a passive avoidance response in the adult rat. PMID- 8247414 TI - Lead-calcium interactions in cellular lead toxicity. AB - The interaction of Pb and Ca with cellular sites depends upon the concentration of free ions present (Pb2+, Ca2+). The ability of Pb2+ to form complexes with simple anions such as Cl- and OH-, the formation of precipitates such as Pb(OH)2 and Pb3(PO4)2, and the ubiquity of Pb as a contaminant in laboratory reagents implies that particular care is needed in order to define the Pb2+ concentration of a solution. The free Pb2+ concentration may be controlled with Pb2+ buffers, and measured with a Pb2+ selective electrode, a fluorescent dye, fura-2, or an NMR indicator, 19F-BAPTA. Pb(2+)-Ca2+ interactions occur in three main situations at the cellular level. Pb2+ and Ca2+ compete at the plasma membrane for transport systems which effect their entry or exit, such as Ca2+ channels, and the Ca2+ pump. Intracellular Ca2+ is buffered to around 10(-7) M by proteins, endoplasmic reticulum and mitochondria. Pb2+ disturbs intracellular Ca2+ homeostasis. Ca(2+) Pb2+ interactions at mitochondria have been described, but other mechanisms have not yet been explored. Increases in intracellular [Ca2+] act as a signal (or second messenger). Pb2+ interacts with a number of Ca(2+)-dependent effector mechanisms, such as calmodulin (a Ca2+ receptor protein which couples to several enzymes e.g., phosphodiesterase, protein kinases), protein kinase C, Ca(2+) dependent K+ channels in the plasma membrane and neurotransmitter release. The actions of Pb2+ on neurotransmission may be relevant to Pb(2+)-induced human neuropathy and encephalopathy. PMID- 8247415 TI - International perspectives of lead exposure and lead toxicity. AB - Three approaches have been used to examine how human body burdens of lead depend on different environments: (1) In paleopathologic studies, lead concentrations have been determined in well-preserved human bones or teeth, and pre-pollution samples generally show lead concentrations of about 1% of current levels in industrialized countries. (2) Geographic comparisons of blood-lead concentrations show low levels in, Nepal, Faroe Islands, and Sweden, while high levels occur in Mexico and Malta; average blood-lead levels may vary by a factor of 10 or more. (3) In analytical epidemiology, major exposure sources have been related to lead levels in blood, by either prospective or cross-sectional design. Increased blood lead concentrations are related to smoking, drinking alcoholic beverages, eating vegetables for dinner, urban residence, and exposure from lead-using industries; average blood-lead values of subgroups within well-defined populations may vary by a factor of 3 or more. The dose-relationships for lead-induced neurotoxicity will depend on the sensitivity of the parameters chosen as indicators of lead exposure and of neurotoxicity. The temporal relationship between lead exposures and the development of deficits must be ascertained. Individual susceptibility and interacting factors must also be taken into account. Differences in addressing these issues impede the comparison between studies. Recently neonatal jaundice has been found to be a risk factor for subsequent neurobehavioral dysfunction in children with a birth weight above 2500 g, but only in children with increased lead exposure. Lead exposure may act in combination with several other factors and result in additive, or synergistic effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247416 TI - Evidence that lead acts as a calcium substitute in second messenger metabolism. AB - Our investigations of the ability of lead to substitute for calcium in several intracellular regulatory events are reviewed in the context of the neurotoxicity produced by this heavy metal. We found that lead activates calmodulin dependent phosphodiesterase, calmodulin inhibitor sensitive potassium channels, and calmodulin independent protein kinase C (PKC). Because of sensitivity in the picomolar range, the activation of PKC by lead may be more biologically relevant than its activation of calmodulin which requires high nanomolar levels of the toxicant. We also found that inhibition of capillary-like structure formation in co-cultures of neural endothelial cells with astrocytes by lead was similar to that produced by phorbol esters, known stimulators of PKC. This functional event supports the hypothesis that PKC activation underlies some aspects of lead neurotoxicity. Taken together these studies implicate second messenger metabolism and protein kinase activation as potential sites for the disruptive action of lead upon nervous system function. These reactions could contribute to the subtle defects in brain function associated with low level poisoning. PMID- 8247417 TI - The nutrition CRSP: what is marginal malnutrition, and does it affect human function? AB - This review of the Nutrition Collaborative Research Support Program (Nutrition CRSP) describes the results of a longitudinal study of the effects of marginal malnutrition on human function in Egypt, Kenya, and Mexico. Growth-stunting occurred soon after birth and was associated with cognitive and behavioral deficits in infancy and childhood. Maternal nutritional status was implicated in this phenomenon. All three population groups had poor dietary quality and multiple micronutrient deficiencies, which were associated with poor function. Energy deficiency was a problem only in Kenya. PMID- 8247418 TI - Does mild zinc deficiency contribute to poor growth performance? AB - Zinc promotes growth in severely malnourished children, and mild zinc deficiency may limit growth in otherwise healthy children. An intervention study involving children aged six to seven years from a poor area of Guatemala failed to show beneficial effects of zinc on height and weight gain, although body fat status was increased. Further studies are needed to clarify the influence of mild zinc deficiency on growth. PMID- 8247419 TI - Dietary fatty acids and protection against experimental cardiac arrhythmias in rats. AB - Sudden cardiac death accounts for about 50% of total coronary disease mortality in westernized industrial countries. The lack of early symptoms for this disorder makes prevention the preferred strategy. In a rat model of cardiac ischemia, dietary n-6 (sunflower seed oil) and n-3 (fish-oil) polyunsaturated fatty acids were shown to protect against arrhythmia compared with saturated fat, with greatest protection observed with fish oil. The frequency of arrhythmia was similar with monounsaturated fat from olive oil and saturated fat. PMID- 8247420 TI - Apo A-IV: a new satiety signal. AB - Chylomicrons have a suppressive effect on food consumption, which is attributed to apolipoprotein A-IV (apo A-IV). This protein is found in cerebrospinal fluid in vivo, and its infusion into the third ventricle of the brain reduces food intake. These findings suggest that this apolipoprotein operates through the central nervous system to regulate food intake. PMID- 8247421 TI - Breast-feeding protects against otitis media. AB - Although exclusive breast-feeding decreases infant mortality and morbidity in developing countries, its protective effects in infants living in industrialized nations have been more difficult to quantitate. A recent study provides strong evidence that exclusive breast-feeding for at least four months decreases the incidence of otitis media in the first year of life. PMID- 8247422 TI - Safety limits for nutrient intakes: concepts and data requirements. AB - The possible quantitative methods calculating safety limits for nutrient intakes are related conceptually to those used to calculate safe limits for exposure to environmental chemicals and to the therapeutic index used to assess the relative safety of drugs. The impact of using a fixed SF has been compared with the use of variable SFs. Of the methods identified, the SRM gives lower limits than does the MPM. However, neither of these methods calculates safety limits below the RDA, even for nutrients with narrow margins of safety. The acceptability criteria for toxicity data for use in identifying safety limits are an issue of major importance and must be resolved before calculated limits may be used to support policy or regulatory decisions. An advantage of adopting a standard formula involving systematically varying SFs to calculate safety limits is that the margin of safety below the expected range of toxicity for each nutrient would be systematic, without having the safety limit for any nutrient regress below its RDA. Once the data acceptability criteria were met, the safety limit would be identified objectively. The confidence in and reasonableness of safety limits, regardless of the method used to define them, will be enhanced if the objectives, data criteria, and the quantitative method have been agreed upon ahead of time by groups responsible for nutrition and health policy. Even with such agreement, the confidence in using such procedures to support policy decisions will be improved by the extent and quality of the data base on toxicity and adverse reactions associated with consumption of excessive levels of the nutrients under consideration. PMID- 8247423 TI - [Evaluation of the quality of nursing care. A concept to get used to]. PMID- 8247424 TI - [Integral management of quality. In life, in death]. PMID- 8247425 TI - [A glance at Brazilian nurses]. PMID- 8247426 TI - [Burkina Faso. Working for the elaboration of nursing care norms]. PMID- 8247427 TI - [The stages of family life cycles]. PMID- 8247428 TI - Conquering professional isolation. PMID- 8247429 TI - The Plunket National Child Health Study: birth defects and sociodemographic factors. AB - AIMS: The Plunket National Child Health Study was set up to examine the health experience of a representative sample of New Zealand children. This paper examines the association of birth defects with the sociodemographic variables of maternal age and education, parity, socioeconomic group, region of domicile and marital status; infant's ethnic group and sex. The effect of maternal smoking on the prevalence of birth defects was also examined. The paper compares the prevalence of birth defects in infants alive at 6 weeks with the reported prevalence at birth as described in various other publications. METHODS: A cohort of 4286 children born in New Zealand during 1990-1 were enrolled in the study. The presence of birth defects in the study population was determined by clinical examination and review of hospital or midwife referrals. All described defects were reviewed and coded according to the ICD-9. RESULTS: The overall prevalence of birth defects in the study was 4.3%. There was a significant association between socioeconomic status and the incidence of birth defect but no other significant sociodemographic variations. Cigarette smoking had no statistically significant effect on the rate of birth defects. There was a marked difference between the rates for certain defects in the Plunket study when compared to Health Department notifications. CONCLUSIONS: Because of the differences between the current study and official Health Department notifications we suggest that criteria for congenital anomaly notification are clarified, and that a second notification takes place at six weeks of age. PMID- 8247430 TI - General practice consultations involving pigmented naevi presented for assessment of malignancy. AB - AIM: To outline the initial general practice care of pigmented naevi presented for assessment of malignancy. METHOD: Fifty six general practitioners completed a brief record for each patient presenting a pigmented skin lesion for assessment of malignancy; records were maintained prospectively for periods over October 1988 to April 1989. 2614 records were received. RESULTS: Numbers of consultations increased sharply with age to peaks at 15-19 for men and 30-44 for women; at ages 20 44 the sex ratio was 1.9 (F/M). Lesions were usually typical (51%) or atypical (24%) moles. 22% of patients were biopsied by the practitioner and 8% were referred, to dermatologists (4%) or surgeons (4%). CONCLUSIONS: The age pattern of presentations is younger than is optimal. General practitioners' decisions and actions are of crucial importance in the effective functioning of melanoma early detection programmes. PMID- 8247431 TI - The quest for normoglycaemia is worth the effort. PMID- 8247432 TI - Short term safety assessment of cilazapril. AB - AIMS: To undertake an event monitoring study of cilazapril in general practice during the early marketing period, to provide some comparisons with other angiotensin converting enzyme inhibitors and to assess the monitoring method. METHODS: The monitoring was undertaken in the Intensive Medicines Monitoring Programme. Cilazapril was prescribed for mild to moderate hypertension in 996 patients at a recommended dose of 2.5-5.0 mg daily. The monitoring period was six months and practitioners were asked to report all adverse events. A reaction profile was prepared and compared with profiles for lisinopril, enalapril and captopril. The chi-square test was applied to differences in proportions. RESULTS: There were 84 (8.4%) reports describing 133 adverse events; 124 (93%) were assessed as reactions. Withdrawals totalled 53 (5.3%). The most common reactions were cough (2.9%), nausea and vomiting (1.3%) and lethargy (1.1%). Cilazapril had a higher proportion of neurological reactions (p < 0.001) (mainly headache) but a lower proportion of skin reactions (p = 0.001) than the other ACE inhibitors. It also had relatively less diarrhoea and there were differences in the patterns of psychiatric reactions. CONCLUSIONS: Cilazapril has a similar reaction profile to other ACE inhibitors but this paper shows differences, some not previously reported, that may assist selection when prescribing. Although there was a high rate of reporting of known adverse reactions, other events were reported at a very low rate and spontaneous reporting is thus confirmed as an unreliable method of monitoring for unexpected adverse reactions. PMID- 8247433 TI - The persistence of anti-hepatitis B surface antibodies to three years of age: is a hepatitis B vaccine booster required? AB - AIMS: To evaluate the persistence of hepatitis B surface antibodies (anti-HBs) after immunisation in early infancy. METHODS: The infants were born to low risk European mothers negative for hepatitis B surface antigen (HBsAg). All the children had received 3 doses of 20 micrograms of recombinant DNA hepatitis B vaccine. RESULTS: One month after the third dose all 92 infants were seropositive. The GMT was 1190 mIU/mL and all but one infant had seroprotective titres above 10 mIU/mL. Three years after the vaccination 91% (59 of 65) children who returned for testing still had measurable anti-HBs titres. The GMT was 32 mIU/mL but 26% (17 of 65) had titres less than 10 mIU/ml. Only one child had serologic evidence of contact with the hepatitis B virus but did not develop the disease. CONCLUSION: This vaccine is safe and effective for at least 3 years. The long term duration of protection from vaccination in early infancy requires further studies. PMID- 8247434 TI - Cystitis and nonsteroidal antiinflammatory drugs: an incidental association or an adverse effect? AB - AIMS: Cystitis, a rare adverse effect of systematically administered drugs, was first reported to be associated with a nonsteroidal antiinflammatory drug, tiaprofenic acid, in 1991. Similar reports of adverse effect of tiaprofenic acid were received by a number of national drug monitoring centres. It was therefore decided to investigate the frequency of cystitis associated with tiaprofenic acid and to see whether this association is typical of tiaprofenic acid or also occurs with other nonsteroidal antiinflammatory drugs. METHODS: The Medicines Adverse Reaction Monitoring Centre has been monitoring drug related events/reactions since 1965. An analysis of the spontaneous adverse reaction reports received at this centre during the period 1965-92 was carried out. RESULTS: Haemorrhagic cystitis was reported to be associated with tiaprofenic acid (n = 3) and indomethacin (n = 3). In addition 11 other reports of haematuria (not associated with any coagulopathy and/or hepatic disorders) in relation to other nonsteroidal antiinflammatory drugs, such as diclofenac, ketoprofen, naproxen and piroxicam were received. Five patients who were rechallenged with the suspect drug, suffered from recurrence of cystitis/haematuria. CONCLUSIONS: In addition to tiaprofenic acid, cystitis appears to be associated with indomethacin and possibly also with other nonsteroidal antiinflammatory drugs. An allergic or immunological mechanism is probably responsible for this reaction. Although our centre received the first report of cystitis in relation to indomethacin in 1966, judging by the number of reports received, this association appears to be poorly recognised. PMID- 8247435 TI - Ask and you shall be given: practice based immunisation coverage information. AB - AIMS: To investigate a method for obtaining immunisation information, and investigate the variation in immunisation coverage levels between practices. METHOD: The Wellington Area Health Board computer identified all children born in the first half of 1990 with a general practitioner in the Wellington district. The general practitioner provided the child's immunisation status, if still with the practice. The characteristics of the practice were tested for association with the immunisation coverage level. RESULTS: The method identified approximately 80% of the target population. Most (97%) of the general practitioners agreed to provide immunisation information on the identified children. Immunisation information was obtained on 841 children, or about 65% of the total cohort. The percentage of the children fully immunised in each practice varied from 0% to 100%. An index of neediness of the children in the practice, was the only identified factor which explained variation between practices. Recall systems were neither necessary nor sufficient for obtaining high coverage in these practices. CONCLUSIONS: The study method could form the basis of a system of gathering immunisation coverage information which assists general practitioners with immunisation recall. PMID- 8247436 TI - Being Maori. PMID- 8247437 TI - Fluoridation and hip fracture. PMID- 8247438 TI - Sexual abuse in the doctor/patient relationship. PMID- 8247439 TI - Anaesthetic drug reaction. PMID- 8247440 TI - Blood oestrogens and high fibre diet. PMID- 8247441 TI - Salmonella infection. PMID- 8247442 TI - Who gets the records? PMID- 8247443 TI - Breaking the code. PMID- 8247444 TI - Positive perceptions. PMID- 8247445 TI - Women dentists: 1993 and beyond. PMID- 8247446 TI - A measure of progress. AB - In the 100-plus years since dental school graduated its first woman, the female presence in dentistry has increased and then decreased. It's on the rise again, causing several organizations to reassess their missions. PMID- 8247447 TI - The fashionable road to dentistry. PMID- 8247448 TI - When a woman chooses dentistry. Interview by Laura Beth Clark. PMID- 8247449 TI - Women: a resource for dental education. PMID- 8247450 TI - Lucy paved the way. PMID- 8247451 TI - Women in dentistry. A point of view. PMID- 8247452 TI - Unequal opportunity. PMID- 8247454 TI - Committed to service. PMID- 8247453 TI - Article in error. PMID- 8247455 TI - The no-fault fee schedule. Two common reimbursement problems. PMID- 8247456 TI - Working length in root canal fillings. PMID- 8247457 TI - Effects of titanium oxide properties on biocompatibility. A preliminary study. PMID- 8247458 TI - On leaving dental practice. The transition. AB - Quit when the choice is yours. Don't do it if your only inner personal identification is that of "dentist"; it might leave you psychologically bereft. PMID- 8247459 TI - The importance of maintaining adequate dental records. PMID- 8247460 TI - Certified radiation safety officers program. The Nassau County experience, 1990 1992. PMID- 8247461 TI - Parenteral sedation education. AB - Since procaine was first synthesized in 1905, local anesthetics have been universally accepted as an effective way to abolish pain. This historical discovery led to advancements in dentistry that continue to this day. PMID- 8247462 TI - Dental survival guide. PMID- 8247463 TI - Conservative management of interstitial pregnancy. AB - Earlier detection of ectopic pregnancies allows the patient and physician the option of conservative management. Conservative surgical management of ampullary ectopic pregnancies has been well described. Traditional management of interstitial or cornual gestation has been by salpingectomy with or without cornual resection or by hysterectomy. In this paper we present a case report of alternative, less radical surgical management and review the literature on conservative surgical and medical management of interstitial pregnancies. PMID- 8247464 TI - Ethical considerations of managed care. AB - Universal health care will soon be here. This paper presents the basic models of managed care as well as a glossary of terms to allow discussion of the basic plans. Three cases are discussed to illustrate the dilemmas of managed care utilizing Dr. Fletcher's ethical model. Basic concepts are derived from these cases to assist physicians to work within the constraints of the new health care systems and enable them to respond to the sometimes unavoidable conflicts between medical benefit and cost containment. PMID- 8247465 TI - Fetal movement in response to vibroacoustic stimulation: a review. AB - Fetal movement in response to vibroacoustic stimulation (VAST) depends on functional maturation of the fetal central nervous system and related to changes in fetal behavior. Researchers have associated the presence of fetal movement with fetal well-being. Use of the maternal perception of sound-provoked fetal movement as a test for predicting fetal health is questionable and may suggest further research. All studies show that there is a clear association between this test and favorable neonatal outcomes. This test is inexpensive and easily obtainable. However, because the test has not been studied on large numbers of patients and maternal reporting at times is biased, its adaptation for clinical use should cautiously wait for clear theoretical advantages. PMID- 8247466 TI - Estimating the age at death in a group of historically unknown adults. AB - Ten diaphysis specimens of femurs and humeri from a total of 4 bodies, three excavated from 14th-15th century graves. (Muromachi period) and one from a tomb of the 7th century (period of ancient burial mounds), were examined using histomorphological methods to estimate their ages. The estimation formula was established using humeri (72 specimen) and femurs (71 specimen) from Japanese aged between 26 and 75 years. The formula was prepared based on a multiple regression analysis of the following 3 items: number of osteons/mm2, mean diameter of osteons, and mean diameter of the Haversian canals. Four estimation formulas were established based on the humeri from all subjects, the femurs from all subjects, the humeri from subjects between 26 and 55, and the femurs from subjects between 26 and 53, respectively. Since the age of the excavated human bones was unknown, we used morphological age estimates (e.g. attrition of teeth, cranial suture closure and similar items) for comparison. Histological and visual estimates correspond in 2 of the 10 cases, but the histological age estimates were higher than the visual estimates in the other cases. In 4 of the 10 cases, age estimates produced by the formulas for younger ages were lower than those by the formulas for all age groups. Thus the formulas for younger ages were not so successful. PMID- 8247467 TI - The internal branch of the superior laryngeal nerve in the dog distributes to the esophageal gland at the pharyngo-esophageal junction. AB - The annular plica (plica anularis) of the cervical esophagus contained mucous glands in dogs. Specimens, impregnated with the silver nitrate, were examined under a dissecting microscope. The internal branch of the superior laryngeal nerve lay fibers in close contact with the terminal portion and excretory duct of glands. PMID- 8247468 TI - Velocities for the Tanner-Whitehouse 2 skeletal maturity in northwest Japanese children. AB - This report describes the velocity of the Tanner-Whitehouse 2 skeletal maturity for some 6,300 Japanese children from Ogi Growth Study 1979-1988. The cubic B spline function fitted the velocities for the Carpal, RUS and 20-bone scores on the smoothed velocity curves. The maturity velocity curves showed single peak around the adolescent period exclusive of a bimodal curve for girls' RUS velocity. Its first peak appeared at 10.9 years and the second, 13.9 years. Just after the first peak their height reached the maximal velocity (11.0 years). The RUS velocity curve for the boys showed the peak maturity velocity at 15.6 years. Their height attained the peak velocity (at 12.9 years) before the RUS maturity did. Although the Japanese children matured earlier for the RUS maturity than Europeans, it was comparable to that of other Asians. The study also showed that the skeletal maturation affected the height growth during the duration of height spurt for the boys, but only during the accelerating period for the girls. PMID- 8247469 TI - Fourier analysis of shape changes in the Japanese skull: occipital view. AB - Photographs (norma occipitalis) of Japanese adult skulls (n = 171) were mathematically described with Fourier analysis to assess shape changes over time. The materials used were adult male skulls excavated from the Kanto District and covered four age-periods; Jomon (2000-1000 BC). Kamakura (14C. AD), Muromachi (15C. AD). and Edo (18C. AD). Mean raw data values, containing both size and shape information of each vector, were compared for the four age-periods. Statistically significant differences among the four age-periods were found using an ANOVA. The form of the Jomon skulls displayed the most difference among the four groups. This trend was especially evident at the parietal margin. Fourier analysis was subsequently undertaken to detect the presence of secular changes in skull form after standardization for size. Once the effect of size was removed, the occipital shape of the Jomon skulls was found to be essentially similar to those of the more recent age-periods.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8247470 TI - Studies on thick sections of the nucleus of mouse Sertoli cells using an electron microscope operating at 300 kV. AB - Examination of the three-dimensional structure of the Sertoli cell nucleus from mouse testes was performed under a high voltage electron microscope operating at 300 kV. Using an en bloc staining method along with fixation by osmium tetroxide and embedding in a mixture of Quetol 651, NSA and MNA, the structures of the nucleus were stained at a high contrast and satisfactory preservation was achieved, thus allowing their study at a high resolution within thick sections. Nuclear components could be observed clearly in 2-3 microns-thick sections of embedded material. Typical three-dimensional configurations of nucleoli and associated bodies were indicated. Thick sections permitted the observation that two or three pernucleolar bodies are usually attached on each side of the nucleoli or form a triangular shape of different sizes of vacuolar structures within the bodies. Stereoscopic observations also revealed overlapping of perinucleolar bodies and nucleoli and suggested the complexity of the components of perinucleolar and intranucleolar chromatin. PMID- 8247471 TI - Distribution of vinculin in the duodenum of chick embryos: immunohistochemical studies at the light and electron microscopic level. AB - Studies on the localization of vinculin and actin in the embryonic chick duodenum between the 9th and 13th days of incubation were performed by light and electron microscopy. In the light microscopic studies, vinculin and actin were localized by immuno-fluorescent double staining with anti-vinculin and rhodamine phalloidin for actin. In the electron microscopic studies, the immunocytochemical distribution of vinculin was examined on Lowicryl K4M thin sections with anti vinculin. Vinculin was prominent near or adjacent to the apical inner membrane of epithelial cells of the previllous ridges in all stages, often located near cell junctions including tight and adherent junctions and the top of microvilli, and weakly in the cytoplasm. Our findings suggested the possible interaction between actin and vinculin in differentiation on the apical side of epithelial cells including the formation of microvilli or terminal webs. PMID- 8247472 TI - A further study on the left-right asymmetry of the planum temporale. AB - By means of a rubbed copy method using India ink and an image-analysis system (IBAS 2000), the areas of 106 left-right plana temporalia were compared using fixed brains of both sexes. The left planum was of a larger size than the right planum in two-thirds of the cases. This result was confirmed statistically by the least-squares analysis of variance method (p < 0.01). PMID- 8247473 TI - The occurrence and fine structure of Merkel cells in the lingual epithelium of the turtle, Clemmys japonica. AB - Electron microscopic examination of the lingual mucosa of the turtle, Clemmys japonica, revealed the occurrence of Merkel cells that shared many morphological features with Merkel cells in other vertebrates. Merkel cells were located exclusively in the basal portion of the epithelium near the taste buds. We also found occasional Merkel cells devoid of nerve contact. Nerve terminals approaching these non-innervated Merkel cells were occasionally found in the connective tissue just beneath these cells. PMID- 8247474 TI - Skinfold compressibility in Japanese university students. AB - It is well known that caliper readings decline after the initial application of the caliper to the skinfold (dynamic compressibility). In addition to this compressibility, there is also a variability of skinfold compressibility at different body sites (static compressibility). To investigate this static variability, a comparison was made between skinfold thickness obtained from using caliper and thickness derived from an ultrasound image (B scan-mode) at sixteen skinfold sites of 96 non-athletic students in good health (45 men and 41 women). The skinfold compressibility is defined as: (uncompressed value-compressed value) x 100/uncompressed value. The patterns created by the plots of skinfold compressibility across the sixteen body sites were similar for both sexes although the inter-site variability is quite large (significant at 0.01 level on ANOVA). Women tend to have greater skinfold compression in the trunk area and less in the limbs as compared with men. This sex difference may be caused by the sex differences of skin thickness and skin tension (subcutaneous space pressure). The marked inter-site variability in skinfold compressibility suggests the need for caution in estimates of fat mass using skinfold calipers. PMID- 8247475 TI - A principal component analysis of morphogenetic field in the root of Japanese dentition. AB - Teeth extracted from Japanese male cadavers were analyzed from the morphogenetic point of view. Variables were buccolingual crown diameter, mesiodistal crown diameter, crown height, root length and total length. Each dimension was analyzed separately by means of principal component analysis with varimax rotation. Components extracted from crown dimensions and total length showed 3 or 4 of 5 underlying components for morphogenetic field, anterior group, molar, premolar, canine and incisor. However, for crown height and root length, the components were less distinct. PMID- 8247476 TI - On the morphology and frequency of Weitbrecht's retinacula in the hip joint. AB - The flattened band reflecting from the fibrous capsule of the hip joint to the head and neck of the femur has been firstly described as "the retinacula" by Weitbrecht in the middle of eighteenth century. In the light of present clinical aspect, the retinacula has been noticed with renewed interests because the nutrient arteries running through this retinacula are one important criterion for orthopedic reduction in the cases of femoral neck fractures. The purpose of the present study is to investigate the morphology and occurrence of this retinacula from anatomical points of view using cadavers. The results show that there are two types of retinacula: a complete band shape and posterior membranous shape. The frequency of retinacula, totaled 94.8 percent in males and 92.5 percent in females. Therefore, the presence and condition of the retinacula should be carefully examined for pre-operative status in cases of femoral neck fractures. PMID- 8247477 TI - Sagittal configuration of spinal curvature line in sportsmen using Moire technique. AB - A separate moire photograph of 380 men's back was taken. Using a micro-computer the sagittal configuration of the spinal curvature line was reconstructed based on the moire fringe on the back. Then the sagittal configuration of this line was compared among groups according to their sports: rugby, soccer, kendo, swimming, yachting, four track and field events (sprinting, distance running, jumping, and throwing), body-building as well as non-athletic men. Distance runners and sprinters had a greater degree of thoracic kyphosis and greater degree of lumbar lordosis, while non-athletes had a smaller degree of both kyphosis and lordosis. Swimmers, bodybuilders, rugby and soccer players had partial lordosis. The degree of lumbar lordosis of swimmers and bodybuilders was less than average while the degree of thoracic kyphosis was ordinary and the degree of thoracic kyphosis of rugby and soccer players was less than average while the degree of lumbar lordosis was ordinary. PMID- 8247478 TI - Using corneal oxygen demand to optimize rigid contact lens design. AB - To evaluate the effects of base curve radius, overall diameter, and axial edge lift on rigid contact lens tear pump efficiency, corneal oxygen uptake rates were measured on six eyes under three conditions: normal open eye, after 5 min of static (without blinking) wear, and after 5 min of dynamic (with blinking once every 5 s) wear. The three parameters were varied one at a time from a standard lens design. Differences in corneal oxygen demands between the static and dynamic rates provided quantitative measures of the tear pump efficiency for each lens design. Tear exchange was found to be most sensitively related to changes in base curve radius, followed by overall diameter changes (66.9% as effective) and axial edge lift changes (64.6% as effective). Design equivalencies (to produce identical tear exchange to a 0.05-mm flattening of base curve toward alignment) were 0.07-mm steepening of the base curve toward alignment, 0.35-mm decrease in overall diameter, or 0.037-mm increase in axial edge lift. PMID- 8247479 TI - Oblique central refraction in spherocylindrical lenses tilted around an off-axis meridian. AB - Thin lens equations accurate to third order are presented for the effective spherocylindrical parameters for oblique central refraction through spherocylindrical lenses that are tilted around an off-axis meridian. This situation occurs in either pantoscopic or faceform tilt for spectacle corrections for oblique astigmats. The thin lens equations appear to be good approximations for oblique central refraction through actual spectacle lenses. PMID- 8247480 TI - The multimeridional apparent frontoparallel plane: introduction of the induced effect. AB - The application of vertical rod stimuli to obtain apparent frontoparallel plane (AFPP) settings is well-known. A geometrical relation based on observation distance, interpupillary distance, and retinal disparity determines the deviation of the setting from the objective frontoparallel plane. Further developments of this procedure have established that a similar relation exists for oblique presentations of parallel rod stimuli, the orientation of the rods being an additional variable. This extension of the AFPP procedure is referred to as the multimeridional AFPP or MAFPP. It permits the determination of aniseikonia in oblique as well as horizontal meridians. Although oblique disparities contain a vertical component, an induced effect is absent as long as parallel rod stimuli are used alone. If an induced effect were present, the MAFPP geometrical relations would be expected to break down. To test this hypothesis, random dot arrays and a row of dots of the same frequency as a control stimulus of parallel lines were presented alone and in combination with the line stimuli. Binocular disparities were induced by a meridional afocal magnifier placed at various axes before one eye. It was found that the dot arrays caused a breakdown of the geometrical relation when presented alone or in combination with the parallel lines. The amount of deterioration varied only slightly with the number and arrangement of dot stimuli but increased as the vertical component of the oblique magnification increased. In addition to proving the main hypothesis, the data provide information pertaining to the MAFPP theory as well as offering some insight into the induced effect. The most important practical implication of the results is that the MAFPP theory can be used to measure unknown retinal disparities only if continuous parallel lines are presented in isolation. PMID- 8247481 TI - The cotton thread test: a brief review and a clinical study of its reliability on Hong Kong-Chinese. AB - Fifty-four asymptomatic Hong Kong-Chinese subjects were recruited and their lacrimation was assessed using a modified cotton thread test (CTT). The CTT was performed once on each subject and, for seven subjects, the test was repeated seven times on different days over a period of 2 weeks to test the reliability of the CTT. The mean (+/- SD) CTT value (length of wetting of thread in millimeters for a 1-min testing period) of this group of subjects was 22.8 (+/- 9.4) mm/min. Repeated measures analysis of variance revealed no significant difference in the CTT values made on different days. The CTT was found to be superior to the conventional Schirmer test in terms of comfort and consistency of results obtained. A brief review of some studies and results reported by previous investigators using the CTT is also presented for comparison. PMID- 8247482 TI - Vision in aging and dementia. AB - PURPOSE: This study was designed to determine the extent of visual deficits in Alzheimer's disease (AD) patients relative to normal aging changes and to patients with other types of dementia. METHODS: Four groups of subjects were tested: patients with probable AD (N = 10), patients with other dementias (N = 10), age-matched controls (N = 11), and young controls (N = 10). Color vision was assessed with the L'Anthony D-15 desaturated color test, contrast sensitivity with the Pelli-Robson chart, and stereoacuity with the RAN-DOT test. RESULTS: Visual changes were found in AD patients that can be accounted for by age-related deficits (color vision), deficits found in other types of dementia (stereoacuity), and deficits specific to AD (low spatial frequency contrast sensitivity). CONCLUSION: We conclude that AD patients show a variety of visual deficits. With our tests, only deficits in contrast sensitivity were specific to AD. PMID- 8247483 TI - Dark focus of accommodation and uncorrected visual acuity. AB - We compared the dark focus of accommodation in 33 healthy children, 19 eyes with good uncorrected visual acuity (1.0 or more) vs. 14 with poor uncorrected visual acuity (below 1.0). The two groups were matched for age and cycloplegic refractive error. Cycloplegic refractive error ranged from +0.15 to +1.75 D in the good visual acuity group and from +0.25 to +1.87 D in the poor visual acuity group. The dark focus was defined as the difference between the refractive error in the dark and the refractive error under cycloplegia. Initially, visual acuity was measured subjectively. Next, refractive error in the dark was measured using the Nidek Autorefractometer AR1600 with its optical target light off. Refractive error under cycloplegia was measured 40 min after the instillation of cyclopentolate hydrochloride using the same autorefractometer. There was a significant correlation between the dark focus and the cycloplegic refractive error (r = -0.53, p < 0.01). Despite an equal refractive error under cycloplegia the dark focus was significantly larger in the eyes with good uncorrected visual acuity than in those with poor uncorrected visual acuity (p < 0.01). Tonic accommodation may influence uncorrected visual acuity. PMID- 8247484 TI - A ballistic evaluation of the impact resistance of spectacle lens materials. AB - The impact resistance of chemically and thermally hardened glass lenses of 2.2- and 3.0-mm thicknesses was evaluated using a ballistic test. Each lens was edged and mounted into a spectacle frame, which was placed on a standard headform. A 6.5-mm steel ball was fired from an air gun at the center of the lens at increasing speeds until the lens broke. The multiple impact data were used to plot cumulative breakage curves. We found that heat-treated photochromic glass and heat-treated crown glass fail at similar missile speeds and that chemical treating considerably improves the impact resistance of crown glass but not of photochromic glass. The poorer performance of photochromic lenses indicates that they should not be prescribed when optimal impact protection is required. Plastic lenses show superior performance. PMID- 8247485 TI - Age, gender, and tear break-up time. AB - We investigated the effect of age and gender on the tear break-up time (TBUT) of 2 groups of normal, asymptomatic subjects [92 Hong Kong-Chinese (HK-Chinese) and 76 Singapore-Chinese (S-Chinese)] using the conventional fluorescein-instillation technique. The mean TBUT (+/- SD) is 6.5 (+/- 4.0) s for the S-Chinese and 7.8 (+/- 2.4) s for the HK-Chinese. The mode was 7.0 s for HK-Chinese and 5.0 s for S Chinese. About 80% of the subjects of each group have TBUT less than 10 s. The TBUT of both groups of Chinese decreases with age. It was found that TBUT was significantly different between the two sexes in the S-Chinese but no equivalent effect was observed in the HK-Chinese. PMID- 8247486 TI - Lacrimal equilibration time (LET): a quick and simple dry eye test. AB - We investigated the benefits of a new test on the lacrimal system for Dry Eye. This additional test to facilitate the diagnosis of dry eye is described and compared with the Schirmer test and with the results of a patient questionnaire. This simple test uses materials available in all optometric practices. An assistant with brief training can administer the test for the optometrist. The patient's monocular distance visual acuity was measured. One drop of Celluvisc was instilled into each eye; a stop watch recorded the time necessary for the patient to regain the predropped visual acuity. Each patient received a standard Schirmer test and answered a questionnaire. Preliminary results from this new test suggest a correlation between a dry eye with patient symptoms and the length of time for return to normal visual acuity after the use of one drop of Celluvisc. Further investigations should be conducted with a larger population using a precise eye drop dispenser. With this new test, a practitioner would have another method to diagnose a dry eye. PMID- 8247487 TI - Ultraviolet disinfection of contact lenses. AB - To evaluate the efficacy of ultraviolet (UV) radiation as a method of disinfecting contact lenses and their storage solutions, we contaminated soft lenses (Bausch & Lomb Optima 38), rigid gas permeable (RGP) lenses (Oxyflow F 30), and their storage solutions with three common bacteria. Escherichia coli (E.c.), Staphylococcus epidermis (S.e.), and Serratia marcescens (S.m.). The storage solutions used were saline solution and RGP conditioning solution. We determined the exposure times to 253.7-nm wavelength UV radiation necessary to disinfect the contact lenses and solutions. The decimal reduction values (D values) found for UV radiation were 10 to 200 hundred times shorter than reported for currently available disinfection systems. For E.c., sterilization was attained after 100 s of exposure. For S.e. and S.m., sterilization occurred after 300 s of exposure. Different contact lens solutions transmit UV radiation to various degrees, with saline solution passing more than 90% of the UV radiation. Thus, our results indicate that UV radiation is an effective and rapid method of disinfecting contact lenses and their storage solutions. PMID- 8247488 TI - Microwave irradiation and soft contact lens parameters. AB - We evaluated the effect of repeated microwave irradiation on soft contact lens parameters to determine if microwave disinfection affects soft lens polymers or parameters. Ten factory-sealed lenses from each of the four FDA categories were placed in Ciba AOsept cases which were filled with 13 ml of sterile unpreserved saline. The cases were placed in a 10-cm diameter circle on the rotating plate of a standard 2450 MHz 650 W microwave oven and exposed to repeated 5-min cycles of microwave irradiation. The lens parameters, including back vertex power, back optic zone radius, diameter, thickness and water content, were measured over a period of 200 cycles to simulate over 6 months of daily disinfection. The postirradiation measurements were compared to baseline measurements taken before microwave exposure. We found statistically significant changes in many lens parameters, but all were small and few were clinically significant. These findings indicate that microwave disinfection does not have a clinically significant effect on the parameters of most unworn soft contact lenses. Further studies are needed to determine the long-term effects of microwave disinfection on worn soft contact lenses. PMID- 8247489 TI - Persistent diplopia associated with strabismus surgery. AB - The primary therapeutic goal for patients with binocular vision problems is comfortable, single, clear, normal binocular vision at all distances and directions of gaze. Strabismus surgery is frequently used to assist in achieving this therapeutic goal. Strabismus surgery is also used for those patients who cannot achieve single normal binocular vision but who have a cosmetically noticeable ocular deviation. However, one of the complications of cosmetic strabismus surgery is persistent diplopia. This possibility must be considered seriously before any cosmetic strabismus surgery is performed. The purpose of this paper is to present a case report of a patient who had diplopia after cosmetic strabismus surgery to emphasize the need for careful consideration of the possibility of persistent diplopia after cosmetic strabismus surgery and how to avoid it. PMID- 8247490 TI - An unusual retinal pigment epitheliopathy endemic to the island of Guam. AB - An unusual retinal pigment epitheliopathy resembling ophthalmomyiasis interna posterior (OMI) is endemic to Guam. Sixteen percent of a sample of the indigenous population were found to have the retinopathy. Fifty-two percent of a group of individuals suffering from amyotrophic lateral sclerosis/Parkinsonism-dementia complex (ALS/PDC) were found to have the same retinopathy. Although the retinal appearance is similar to that caused by a subretinal parasite, a definite etiology has not been identified. Three case reports are presented as samples of the clinical presentation of the condition. PMID- 8247491 TI - Combined hamartoma of the retina and retinal pigment epithelium. AB - Combined hamartoma of the retina and retinal pigment epithelium is a rare, congenital fundus lesion which typically presents with a history of reduced acuity and/or the onset of strabismus. Clinically, the lesion is elevated, with a variably pigmented outer portion and a lighter central core of dilated tortuous vessels and gliosis. This benign lesion may be confused with more threatening conditions such as malignant melanoma or retinoblastoma. An appropriate diagnosis through clinical appearance and fluorescein angiography is imperative to prevent unnecessary enucleation. The authors report one case of combined hamartoma of the retina and retinal pigment epithelium in a patient presenting with a complaint of decreased vision. PMID- 8247492 TI - National Eye Institute: 25 years. PMID- 8247493 TI - The relationship between birthdate and primary grade school achievement. PMID- 8247494 TI - The specialty of maxillofacial radiology. PMID- 8247495 TI - Unusual presentation of a chondromyxoid fibroma of the mandible. PMID- 8247496 TI - Bonded arch bars in oral and maxillofacial surgery. An update. AB - From 1987 to 1992, 127 cases that involved maxillomandibular trauma, reconstruction, and various oral surgical procedures were managed with arch bars bonded to the teeth. The aim of this article is to present the method of application of the arch bar and to report the results of management with this technique stressing its use with 99 trauma patients. PMID- 8247497 TI - Eosinophilic granuloma of the mandibular condyle. A case report and management discussion. AB - The occurrence of eosinophilic granuloma in the mandibular condyle has been reported in only three cases. An additional case of eosinophilic granuloma of the mandibular condyle is reported. The lesion presented was treated with condylectomy and surgical curettage. There was no recurrence or soft tissue involvement on annual postoperative examination. Secondary reconstruction was planned but was not required. PMID- 8247498 TI - Osseous choristoma of the tongue. Report of two cases. AB - Osseous choristoma of the tongue is a relatively rare lesion with 53 cases previously published in the literature. In this article, two additional cases are described; one in a 53-year-old woman located close to the foramen caecum, and the other in a 18-year-old woman on the left of midline and just anterior to the circumvallate papillae that was observed over a 13-year period. PMID- 8247499 TI - Salivary antimicrobial proteins in patients with Crohn's disease. AB - The aim was to study the level of salivary proteins with antimicrobial properties in persons with Crohn's disease. Twenty-five patients were recruited, 13 with ongoing symptoms (acute group) and 12 free of clinical signs of the disease at the time of the investigation (nonacute group). A control group matched to the nonacute group was also included in the study. Unstimulated and stimulated whole saliva samples were collected, and the secretion rates estimated. Unstimulated saliva was analyzed for concentrations of total protein, peroxidase, thiocyanate, slgA, lactoferrin, lysozyme, and for specific bacteria aggregation ability. Numbers of mutans streptococci and lactobacilli in saliva were determined, and dental caries status was examined. No differences were found among the groups regarding salivary flow rate, total protein, or any of the antimicrobial proteins. However, three patients with Crohn's disease had no detectable slgA in saliva compared with none in the control group. The lactobacillus count and the number of decayed tooth surfaces were higher in the nonacute group than in the control group. PMID- 8247500 TI - Oral carriage of Candida in healthy and HIV-seropositive persons. AB - The prevalence of oral colonization with Candida species was studied in 28 HIV seropositive and 28 healthy persons. Candida was cultured from 75% and 68% of HIV positive and control persons, respectively, with a significantly higher density carriage in the HIV-seropositive group. Positive smears were seen in 39% of all patients. Candida albicans was the most frequently isolated species with Biotype 1 accounting for 56% of the isolates. Resistance to antifungal agents was seen in Candida strains isolated from both groups. PMID- 8247501 TI - Multiple central giant cell lesions with a Noonan-like phenotype. AB - A small number of patients with the phenotypic features of Noonan syndrome have also developed giant cell lesions of the jaws similar to those seen in cherubism. This case report describes an individual with the features of the recently described Noonan-like/multiple giant cell lesion syndrome. Previously reported cases and issues relative to diagnosis and management are considered. PMID- 8247502 TI - Chemoprevention of tongue carcinogenesis in rats. AB - The inhibitory effects of vitamin A, selenium, butylated hydroxytoluene, and their combinations on tongue carcinogenesis induced by 4-nitroquinoline 1-oxide were examined in Sprague-Dawley rats. After 4 months of administration of 4 nitroquinoline 1-oxide, a wide range of lesions from hyperplasia to early invasive carcinoma was seen. The lesions were fewer and the carcinomas were less advanced in the rats given preventive treatment. The chemopreventive effect of vitamin A plus butylated hydroxytoluene was most effective. After 7 months of 4 nitroquinoline 1-oxide administration, the incidence, the macroscopic diameter, and the microscopic depth of invasion among carcinomas were smaller in the chemopreventive groups than in the nonchemopreventive group. We conclude that vitamin A, selenium, and butylated hydroxytoluene inhibit tongue carcinogenesis induced by 4-nitroquinoline 1-oxide in rats. The inhibitory effect of combined chemopreventives is not always superior to that of each chemopreventive agent used alone. PMID- 8247503 TI - Multiple dens invaginatus. A case report. AB - A case of dens invaginatus in all four quadrants is reported. Although it is seen mostly in maxillary permanent incisors, mandibular involvement is rare. In this article, in addition to the involvement of the maxillary lateral incisors, the involvement of the right and left mandibular premolars has been presented as well. PMID- 8247504 TI - Intentional replantation of endodontically treated teeth. AB - This article reports 31 cases of intentional replantation with an overall success rate of 80.6% with six recorded failures. Survival times varied from 1 day to 22 years. A second mandibular molar that failed after 3 weeks was replanted successfully a second time with no signs of failure after a 46-month follow-up period. Two cases of intentional replantation of deciduous molars are also reported. We suggest that intentional replantation reduces adverse outcomes and should be considered as another treatment modality and not as a procedure of last resort. This procedure is recommended as the treatment of choice in the following instances: for lower second molars, for single-rooted teeth, and lower first molars when there is difficulty in accessibility to perform apical surgery; when the mental foramen is superimposed over the apex of the premolars; when the molar apex is in proximity to the mandibular canal; when patients object to periradicular surgery; when failures occur after apical surgery, or when surgery would create a periodontal pocket as a result of extensive bone removal. Periradicular surgery with a retroseal is more advantageous for first molars when roots are more curved, widespread, and are more prone to fracture compared with second molars whose roots are more tapered and close together. PMID- 8247505 TI - MR imaging of the TMJ. Decreased signal from the retrodiskal tissue. AB - In clinical work we have observed decreased signal intensity from the retrodiskal tissue in some patients with temporomandibular joint disk displacement. The frequency and clinical significance of this observation is unknown. To document the frequency and to assess its clinical significance we analyzed magnetic resonance images of the temporomandibular joints of 69 patients with temporomandibular joint displacement and 28 asymptomatic volunteers. All the subjects had a clinical examination, and their level of pain was recorded. Decreased signal intensity from the retrodiskal tissue was seen in 16 joints in the patients (12%). None of the asymptomatic volunteers exhibited this feature. It was seen in one joint with disk displacement with reduction, in nine joints with disk displacement without reduction, and in six joints with disk displacement without reduction and arthrosis. It was not seen in any joint with superior disk position. We could not appreciate an association between the decreased signal intensity from the retrodiskal tissue and the level of pain because this feature was seen in both painfree and painful joints. It was concluded that decreased signal intensity from the retrodiskal tissue was most frequently associated with later stages of disk displacement and its clinical significance remains unclear because there was no appreciable correlation to patient symptoms of pain. PMID- 8247506 TI - Contour mapping of the TMJ temporal component and the relationship to articular soft tissue thickness and disk displacement. AB - The osseous architecture of central sagittal histologic sections of the temporal component of 51 temporomandibular joints of young adults at autopsy was studied to determine if this predicted the thickness of the overlying articular soft tissue and disk displacement. Geometric groupings of fossa-eminence shapes were generated using a hierarchical cluster analysis of the osseous fossa-eminence sigmoid curve, size, and slope. Six cluster groups were evolved and tested for relationship to soft tissue thickness measured at the eminence crest, mid-point of the eminence slope, closed pack location of the condyle, the inflection point, and the depth of the fossa. Soft tissue thickness at the inflection point and depth of the fossa was remarkably constant between cluster groups. Increased soft tissue thickness at the eminence crest and lower part of the posterior slope was weakly correlated to a flatter eminence slope and curve, explaining 10% to 20% of the variance (r2). There were no relationships between the parameters studied to the fossa curvature. An ANOVA showed no statistical difference in the posterior slope angle between the categories of disk position (p = 0.715) or to the six cluster groupings. The results suggest that the osseous contours seen on radiographs may not accurately predict the actual articular surface of the temporal component. PMID- 8247507 TI - Relationship of articular soft tissue contour and shape to the underlying eminence and slope profile in young adult temporomandibular joints. AB - This study examined whether the overall shape of the articular soft tissue overlying the posterior slope and articular eminence of the temporal bone could be predicted by the underlying osseous contour in a histologic model of 51 central sagittal sections of young adult temporomandibular joints. Articular soft tissue and bone contours were traced, and osseous landmarks identified on the basis of joint geometry. Soft tissue thickness measurements were made under low power light microscopy. Seven categories of articular soft tissue pattern were identified. The soft tissue uniformly followed the osseous contour in only one (14%). A progressive increase in soft tissue thickness from the middle of the posterior slope to the articular crest was the most common pattern (35%) but did not describe most of the sample that was more asymmetric. Pattern was poorly predicted by the shape and slope of the temporal bone outline or by dental factors that describe anterior guidance and did not relate to disk displacement. The articular soft tissue compensated for flatter eminence slopes and osseous irregularities and maintained an intact surface. This study has clinical implications for radiographic interpretation of disk space, condyle translation pathways, and the integrity of the functional articular surface. PMID- 8247508 TI - MRI effects on craniofacial size and crown-rump length in C57BL/6J mice in 1.5T fields. AB - Previous studies have shown magnetic resonance imaging to be potentially teratogenic for eye development. An investigation was undertaken to ascertain the effects of 1.5 Tesla magnetic resonance imaging fields on crown-rump length and craniofacial perimeter, two less sensitive teratologic end points of the C57BL/6J mouse. A sham control and two experimental groups of dams (N = 12, 11, 12) placed in different magnet locations were exposed to magnetic resonance imaging fields under clinically realistic conditions. A T-2 weighted spin-echo technique of 36 minutes duration was used on each dam. Crown-rump length was chosen because it is a standard measure of embryotoxicity. Craniofacial perimeter was evaluated because of its relationship to the anterior neural plate, the same region in the developing embryo that gives rise to the eyes. Magnetic resonance imaging fields were found to produce crown-rump and craniofacial perimeter measures smaller than control animals (p < 0.05) when exposed at the isocenter (p = 0.038 and 0.008 respectively). Groups exposed to magnetic resonance imaging fields at the magnet entrance showed p values of 0.004 and 0.053 respectively. The results confirmed magnetic resonance imaging teratogenicity in the C57BL/6J mouse and demonstrates the need for further experimental investigation to ascertain the clinical safety of magnetic resonance imaging for humans. PMID- 8247509 TI - Bacterial contamination of dental radiographic film. AB - Three types of paper-covered and one type of plastic-covered Kodak dental film were used to determine if bacteria could penetrate the coverings and contaminate the inner film. Films from each group were immersed for 30 or 120 seconds in high concentrations of known bacterial suspensions with or without 10% sterilized calf serum added to the incubation media. The plastic-covered film effectively excluded all bacteria, whereas the paper-covered film showed contamination with all organisms even at 30 seconds. Increasing the viscosity of the incubation medium with calf serum decreased the level of contamination. PMID- 8247510 TI - Radiographic interproximal angulations: implications for rotational panoramic radiography. AB - Axial radiographs were taken on 160 subjects at the screening clinic of the Dental School, University of Texas Health Science Center at San Antonio. Intermeatal and midsagittal lines, together with coordinate axes and polynomial curves that represent the average dental arch form, were overlaid on each radiograph. Coordinate references for interproximal tangents at 3154 contacts along the average dental arch form were digitized and angulations between the arch form and midsagittal plane calculated. Interproximal angulations at the average arch form were found to vary greatly but the average fluctuated around 90 degrees over the length of the arch. Optimal beam angulations were shown to be considerably different from central ray angulations of current panoramic machines, and although most machines demonstrated favorable interproximal beam angulation in the anterior region, discrepancy in the premolar region ranged from 15 degrees to over 40 degrees. Overall the OrthOralix SD (Gendex Dental Systems, S.r.l., Monza, Italy) orthogonal projection and PM 2002 CC (Planmeca Inc. Helsinki, Finland) deviate least from optimal interproximal angulation over most of the dental arch. However, the Planmeca gives a better angulation in the canine/premolar region, whereas the OP5/10 (Palomex Instrumentarium Corp., Helsinki, Finland) is comparable with the OrthOralix SD in the molar region. PMID- 8247511 TI - Unusual case of root resorption of permanent incisors. PMID- 8247512 TI - [Human cytomegalovirus, its significance in immune deficiency states, laboratory diagnosis, therapeutic possibilities]. AB - The authors report on the virological findings of 59 transplant recipients. The following procedures were used for the detection of human cytomegalovirus (HCMV) infection: detection of antiviral antibodies by ELISA, the detection of virus coded antigens in the patients' leucocytes (HCMV antigenemia test), "accelerated" virus isolation using immunofluorescence (IF). Serial examinations revealed the HCMV infection in 12 patients following organ transplantation. The antigenemia test proved to be positive in all cases. Two third of the cases suffered from viremia. The virus specific serology possess diagnostic value only in every second acute illness. Since the antigenemia test used to be successful in the earliest phase of acute illnesses, the chance of effective chemotherapy can be increased significantly. The virus serological examinations are of essential importance during the pretransplantation screening of donors and recipients. The "accelerated" procedure of virus isolation experiments indicates the presence of infective HCMV within 1 to 4 days. Transplant recipients obtain new life perspectives, nevertheless, the modern diagnostic procedures may only support the prevention of life-threatening virus infections under the conditions of immunosuppression. The excellent mutual cooperation of the clinicians and diagnostic virologists seems to be at least as important condition of successful transplantation medicine as the high technology in surgery and clinical diagnostics. PMID- 8247513 TI - [Lymphoid infiltrates in the lung]. AB - The authors reviewed material of 10 year period (1980-1990) of the Department of Pulmonology, Albert Szent-Gyorgyi University of Medicine, Deszk, Hungary, and selected 14 patients from the files who considered to belong in one of the lymphoproliferative conditions (4 low grade and 4 high grade lymphomas of B-cell type, 1 angiocentric, 1 mediastinal lymphoblastic non-Hodgkin's lymphoma, 3 Hodgkin's lymphoma cases and 1 pleural pseudolymphoma). Every patient admitted with prominent pulmonary symptoms. The diagnoses were based on histology and immunohistochemistry of tissue samples and autopsy. One high grade B-cell and the angiocentric malignant lymphoma proved to be primary pulmonary process. No specific radiomorphological signs were found, which could be characteristic for the pulmonary lympho-reticular infiltrations and also to distinct the primary and secondary ones. The lung infiltrations in the most of the non-Hodgkin's lymphoma cases with low grade malignancy appeared imitating tuberculosis, while the high grade group and Hodgkin's lymphomas displayed confusion with any primary or multiple tumors. PMID- 8247514 TI - [The place of percutaneous transluminal angioplasty in the management of obliterative vascular diseases of the lower extremities. Femoro-popliteal arterial angioplasty]. AB - Authors treated 331 patients having symptoms of femoropopliteal stenoses or occlusions with transluminal angioplasty from 1983. They performed 186 dilatations and 189 recanalizations. Procedures were preceded by medicinal preparation. The primary technical success rate was 94.6% after dilatation, and 79.9% after recanalisation. In case of stenosis the success rate did not show significant connection with the length of the dilated segment but in case of occlusion longer than 6 cm the success rate decreased. After procedure 87.2% of the patients became asymptomatic, or improved. 21 (5.6%) complications (7 inguinal hematomas, 6 emboles, 6 thromboses, one perforation of arterial wall) occurred and twelve of them required surgical intervention. One patient died after angioplasty due to cardiac failure. Authors are of the opinion that femoropopliteal occlusions and stenoses shorter than 6 cm should be treated by angioplasty. They suggest making balloon catheter dilatation for multiple stenoses even they are longer than 6 cm. PMID- 8247515 TI - [Bacillary angiomatosis]. AB - In a 78 years old patient with chronic lymphoid leukemia, diabetes mellitus a cat scratch induced disseminated angiomatous papules were observed. In the lesions great number of bacilluses were observed with light -and electron microscope. As a result of antibiotic treatment the lesions regressed without trace. This opportunist infection resulting general symptoms as well, may be regarded as a cutaneous manifestation of immunodeficiency. The adequate antibiotic treatment depends on the exact diagnosis. PMID- 8247516 TI - [Medical indications for HIV serological tests]. PMID- 8247517 TI - [Revised definition of AIDS cases and reclassification of HIV infections in adolescents and adults]. PMID- 8247518 TI - [Relation of angiographic changes to clinical data and risk factors in patients with arteriosclerosis obliterans]. AB - The arteriographical changes of 66 patients with peripheral arterial obliterative disease and the relationship between the observed alterations and certain clinical and laboratory data as well as some risk factors were analysed. A score system was used to quantify the arteriosclerotic lesions. The arterial alterations of the examined 90 extremities were studied in the pelvic (common and external iliac arteries), femoral (common, superficial, deep femoral and popliteal arteries) and crural (anterior and posterior tibial arteries) regions. Pathomorphological changes in 433 of the investigated 720 arteries were shown, from among them plaque in 11.8%, stenosis in 22.3%, total occlusion in 26% were proved. Most frequently (89%) the superficial femoral artery was affected. As to the single vessels, the superficial femoral and posterior tibial arteries, while among the zones the femoral region proved to be most serious radiologically. The degree and extent of vessel lesions were related to age. The morphological severity was basically concordant with the results of clinical investigations (walking distance, ergometry, Doppler index). The separately examined risk factors (smoking, hypertension, diabetes, hypercholesterolemia, hypertriglyceridemia) were not related consequently to the radiological changes, but in certain combinations, as heavy smoking with hypertension and heavy smoking with diabetes, the relationship proved to be statistically significant. PMID- 8247519 TI - [Complex study of selenium levels in healthy subjects in Hungary]. AB - Selenium supply of different populations might vary substantially according to the geographical location and eating habits. The authors pioneered in surveying the selenium supply of the Hungarian population in diverse parts of the country. The former was achieved by the study of multiple parameters characteristic of the human body's selenium status. Selenium levels were determined in sera of blood donors and in toe and fingernails of healthy village and city populations. Measurements in sera and in nails were performed by electrothermic atomic absorption spectroscopy and by flourimetric method, respectively. The average selenium concentration in sera was 0.70 micromol/liter, in fingernails was 0.61 mg/kg, in toenails it was 0.56 mg/kg. The average selenium concentration was significantly higher in the south-eastern and significantly lower in the south western regions of the country compared to the northern countries. Similar regional differences could be observed in the average selenium content of both toe and fingernails. According to the findings, selenium deficiency could be verified even in the majority of persons with the highest selenium levels. PMID- 8247520 TI - [Generalized secondary amyloidosis in rheumatoid arthritis]. AB - Generalized (systemic) secondary amyloidosis was a post mortem finding recorded in 28 patients (25.2%) by amyloid specific Congored staining according to Romhanyi, from 111 randomized autopsy cases with rheumatoid arthritis. Amyloidosis was present most frequently in the gastro-intestinal tract, the heart, kidney, thyroid gland, spleen and in the suprarenal glands. The quantity of amyloid deposits is usually the highest in the organs that are most frequently have deposits. The amyloidosis is a progressive cumulative process. The conclusion is that the organs where there are frequently high quantities of amyloid is where the deposits begin and where there are infrequently any deposits or low quantities is where the deposits develop later. Biopsy of the most frequently involved organs are suggested for early diagnosis of amyloidosis. Comparing the laboratory parameters (at the last hospitalisation), the amyloidotic patients were anaemic, with low haemoglobin level (p < 0.05), the renal function significantly decreased, the carbamid (p < 0.001), and kreatinin level high (p < 0.05), with frequent proteinuria (p < 0.0001). PMID- 8247521 TI - [Experience with rapid gallstone dissolution]. AB - The authors describe results of rapid percutaneous litholysis in 26 patients with gallbladder stones. The stones were completely dissolved in 17 cases and became free of symptoms 7 patients. The mean hospitalization time was 3 days, in eight cases only 1 day. PMID- 8247522 TI - [Diagnosis of Verner-Morrison syndrome (VDHA) and its treatment with sandostatin an streptozocitin]. AB - A case report is presented of a man with Verner-Morrison syndrome of extreme severity, caused by an unresectable pancreatic VIPoma. The pathological role of vasoactive intestinal polypeptide (VIP) is discussed in the pathogenesis of Watery Diarrhoea, Hypokalaemia, Achlorhydria (WDHA) syndrome. The authors describe the typical symptoms of the syndrome and provide a diagnostic and therapeutic strategy. Plasma level of VIP was determined by the authors' own VIP RIA method. Administration of a long acting somatostatin analogue, octreotide (Sandostatin, Sandoz) at a dose of 100 micrograms daily, decreased the plasma level of VIP from about 55 to 38 fmol/ml, which was associated with complete regression of the diarrhoea. Due to the 'escape phenomenon' the dose of Sandostatin was gradually increased and finally completed with streptozotocin (Zanosar, Upjohn) administration, which was repeated every 8 weeks. The combination of Sandostatin and streptozotocin resulted in complete regression of diarrhoea and substantial diminution of the tumour mass. The patient displayed a weight gain and returned to normal life. PMID- 8247523 TI - [Fatal myocardial changes in systemic lupus erythematosus]. PMID- 8247524 TI - Phosphorylation sites mapping in the N-terminal domain of c-myc modulate its transforming potential. AB - The nuclear proto-oncoprotein c-Myc is involved in the regulation of cell growth and differentiation. c-Myc is phosphorylated at multiple sites in vivo, two of which we have identified near the amino terminus. In chicken Thr-61/Ser-65 are phosphorylated, as are the comparable positions, Thr-58/Ser-62 in human c-Myc. These residues are located within a domain that is implicated in transactivation and is important for the transforming potential of the protein. Furthermore, these phosphorylation sites or nearby amino acids are frequently mutated in v-myc and in several c-myc genes from Burkitt's lymphoma cells. In vitro these two phosphorylation sites can be modified by glycogen synthase kinase 3 and mitogen activated protein kinase. To address their biological importance we mutated these amino terminal phosphorylation sites separately and together. Stably transfected Rat1A cells expressing the mutated proteins have an increased growth potential in soft agar compared to wt-c-myc transfectants. These altered transformation characteristics indicate that Myc function may be negatively regulated by the amino terminal phosphorylation. PMID- 8247525 TI - Identification of casein kinase II phosphorylation sites in Max: effects on DNA binding kinetics of Max homo- and Myc/Max heterodimers. AB - Myc proteins have been implicated in the regulation of cell growth and differentiation. The identification of Max, a basic region/helix-loop helix/leucine zipper protein, as a partner for Myc has provided insights into Myc's molecular function as a transcription factor. Recent evidence indicates that the relative abundance of Myc and Max is important to determine the level of specific gene transcription. In this report we have identified two major in vivo phosphorylation sites in Max (Ser-2 and -11) which can be modified in vitro by casein kinase II (CKII). Phosphorylation of these sites modulates DNA-binding by increasing both the on- and off-rates of Max homo- as well as Myc/Max heterodimers. In addition, our data indicate that the steady state binding of the shorter version of Max (p21) to DNA was similar yet its rate of dissociation faster than that of longer version of Max (p22). These data argue that different Max complexes have different kinetic properties and that these can be modified by CKII phosphorylation. We propose this as an important biological mechanism by which different dimeric complexes can exchange with varying efficiencies on DNA, thereby responding to changes in cell growth conditions. PMID- 8247526 TI - Early response gene signalling cascades activated by ionising radiation in primary human B cells. AB - We have used a panel of 13 protein kinase C-responsive immediate early gene probes to dissect the cellular signalling pathways activated by ionising gamma radiation in primary human B cells. Of these 13 genes, a delayed transient induction was observed for only 8: c-fos, c-jun, jun-B, jun-D, c-myc, ergI/krox 24 and two 'anonymous' genes, 3L3 and 19A. Expression of c-myc and c-fos mRNAs was paralleled by the appearance of their encoded proteins suggesting that these oncoproteins may couple radiation signalling to cellular responses. Of three protein kinase C-coupled transcription factors examined by gel retardation assay, (AP1, NF kappa B, EgrK/Krox24) only NF kappa B and, to a lesser extent, AP1 was stimulated in response to irradiation. These observations are not obviously compatible with a simple model invoking protein kinase C in radiation signalling in primary B cells and suggest that the pleiotropic effects of ionising radiation on this cell type are mediated through a distinct cellular signalling cascade. PMID- 8247527 TI - Molecular characterization of a variant Ph1 translocation t(9;22;11) (q34;q11;q13) in chronic myelogenous leukemia (CML) reveals the translocation of the 3'-part of BCR gene to the chromosome band 11q13. AB - We performed cloning and sequence analysis of translocation junctions at 11q- and 22q- (Ph1) chromosomes and the corresponding germline DNAs of a variant Ph1 positive CML with t(9;22;11)(q34;q11;q13). Southern blot analysis using probes for different regions of bcr mapped the translocation break near the 5'-side of bcr exon 4. Cloning, Southern blot analysis and restriction map analysis of both bcr fragments showed that the part of bcr 3'- to the translocation break moved to 11q13. Sequence analysis of the translocation junction on the Ph1 chromosome showed that the translocation break occurred 63 bp upstream of exon 4. Compared to the germline sequence, bcr sequence from the translocated partners showed deletion of seven basepairs at the site of translocation. A probe derived from the 5'-region of the clone isolated from the 11q- chromosome identified clonal rearrangements in the leukemic DNA. Restriction map and sequence analysis showed that this clone consisted of the 3'-half of the glutathione S-transferase Pi (GST Pi) gene and the 3'-part of bcr. We identified two point mutations in the GST-Pi allele involved in translocation. Northern blot analysis showed that the GST-Pi gene was expressed in the leukemic cells at blast crisis but not at chronic phase; however, no fusion mRNA between GST-Pi and bcr was identified. We did not find any sequence homology between 11q13 DNA and 22q11 DNA around the translocation breakpoints; however, sequences homologous to ALU repeats were identified close to the sites of translocation breaks at 22q11 and 11q13. This study supports our hypothesis that variant Ph1 translocations may occur as primary cytogenetic changes similar to the classical Ph1 translocations. PMID- 8247528 TI - RAS is required for epidermal growth factor-stimulated arachidonic acid release in rat-1 fibroblasts. AB - Previous studies have provided suggestive evidence for an interaction between ras activation and signalling pathways involved in agonist-stimulated arachidonic acid release in a variety of cell systems. In order to clarify this interaction, we have measured epidermal growth factor (EGF)-stimulated arachidonic acid release in rat-1 fibroblasts transfected with the N-17 dominant negative mutation of ras. Cells transfected with the N-17 ras mutant, display a markedly attenuated arachidonic acid-release response to EGF, compared to sham-transfected and non transfected cells. In contrast, the response to phorbol myristate acetate (PMA) was not attenuated in the N-17-mutant expressing cells. No differences were detected between sham-transfected and N-17 mutant expressing cells in levels of immunodetectable EGF receptor, cytosolic phospholipase A2 or mitogen-activated protein (MAP) kinase. Attenuation of EGF-stimulated arachidonic acid release in the N-17 mutant expressing cells, was accompanied by a marked diminution in EGF stimulated tyrosine phosphorylation of MAP kinase. We conclude that the signalling pathway involved in epidermal growth factor-stimulated arachidonic acid release is similar to the signalling pathway for mitogenic responses to epidermal growth factor and requires ras activation, likely followed by a downstream cascade of kinases eventuating in MAP kinase activation. PMID- 8247529 TI - Over-expression of transforming growth factor alpha antagonizes the anti tumorigenic but not the differentiation actions of retinoic acid in a human teratocarcinoma cell. AB - All-trans retinoic acid (RA) treatment of the multipotent human teratocarcinoma (TC) cell line NTERA-2 clone D1 (abbreviated NT2/D1) induces a neuronal phenotype and other cell lineages. NT2/D1 cells basally express transforming growth factor alpha (TGF-alpha) mRNA and secreted protein. After RA-treatment TGF-alpha expression is markedly reduced. This decline in TGF-alpha expression accompanies the induction of the neuronal phenotype and a marked reduction of tumorigenicity in athymic mice. This suggested a causal link between reduced TGF-alpha expression and the induced differentiation or loss of tumorigenicity of these RA treated TC cells. To evaluate this possibility, an RA-refractory NT2/D1 subclone was analysed. This subclone, designated NT2/D1-R1, failed to induce differentiation or to decrease TGF-alpha expression despite RA treatment. To further explore the relationship between TGF-alpha expression and RA actions in this human TC cell, a TGF-alpha cDNA was stably transfected and expressed in NT2/D1 cells. RA-treatment of independently obtained TGF-alpha over-expressing clones and a representative control transfectant only expressing the neomycin resistance gene produced a neuronal phenotype similar to parental NT2/D1 cells as assessed by morphologic, immunophenotypic, and gene expression markers of differentiation. RA-treatment of these clones also induced a G1 arrest similar to parental cells. However, only the TGF-alpha over-expressing clones that secreted high levels of TGF-alpha protein into the conditioned media before and after RA treatment still developed tumors in athymic mice despite prior exposure to these cells to RA. This finding demonstrates that TGF-alpha can inhibit the anti tumorigenic effects of RA in human TCs. Thus, over-expression of a single growth factor that normally declines with RA treatment antagonizes the anti-tumorigenic but not the differentiation actions of RA in this human tumor cell. PMID- 8247530 TI - The chromosomal translocation t(X;14)(q28;q11) in T-cell pro-lymphocytic leukaemia breaks within one gene and activates another. AB - Chromosomal translocation t(X;14)(q28;q11) has been observed in patients with pro lymphocytic T-cell leukaemia (T-PLL). In two cases of T-PLL, one of which was associated with Ataxia telangiectasia (AT), the chromosomal break occurred in two different introns of a gene c6.1A, located at the Xq28 locus. Fusion transcripts, consisting of 5' sequences of c6.1A and the TCR alpha constant (C) region, were expressed at high levels in the leukaemic cells from both patients, but in only one case did this fusion generate an in-frame c6.1A-C alpha mRNA. However, the breaks within c6.1A seem to affect another gene, c6.1B, which is transcribed from the same CpG rich island as c6.1A but in the opposite transcriptional orientation. The c6.1B gene is not damaged by the translocation but is transcribed in both T-PLL cases. Furthermore, c6.1B may lack protein coding capacity and thus this translocation might result in a novel mechanism in tumorigenesis. In any event, this is the first cloned gene which is implicated in pathogenesis of chronic/pro-lymphocytic leukaemia of the T-cell lineage. PMID- 8247531 TI - E1A 12S in the absence of E1B or other cooperating oncogenes enables cells to overcome apoptosis. AB - Neonatal rat kidneys are still undergoing growth and differentiation. Within 24 h after placing these cells in culture, programmed cell death has been activated. This is evidence by intranucleosomal DNA digestion. Cellular DNA synthesis and proliferation cease between 48 and 72 h after plating and the epithelial cells begin to die. The adenovirus E1A 12S gene, in the presence or absence of the E1B gene products, restarts the proliferation program of these cells, retaining their differentiated states. 12S can accomplish this in the presence and absence of serum. Expression of the E1A 13S gene results in cellular DNA degradation due to necrosis, but not apoptosis, brought on by the expression of the 13S-dependent viral genes. Those cells that retain the 12S sequences escape apoptosis and proceed to become immortal. In addition, 12S enables Madin Darby canine kidney (MDCK) epithelial cells to overcome TGF-beta 1 induced inhibition of proliferation and apoptosis. Thus, E1A 12S may be an anti-apoptotic gene. PMID- 8247532 TI - Wild type p53 functions as a control protein in the differentiation pathway of the B-cell lineage. AB - An analysis of cell lines representing different stages of the B-cell differentiation pathway indicated that about 50% of the cell lines examined expressed exclusively wild type p53 protein. These lines therefore offer a convenient system to study the involvement of p53 in cell differentiation. When 70Z/3, a pre-B cell line which expresses wild type p53, was treated with the differentiation inducer lipopolysaccharide (LPS), it was seen that increased levels of p53 mRNA preceded specific changes in kappa (kappa) immunoglobulin expression. This increased expression of kappa specific mRNA, which was evaluated by specific PCR analysis, was blocked following transfection with mutant p53 coding plasmids. Treatment of 13A60, another cell line which endogenously expresses wild type p53, with LPS caused a secretion of IgA antibodies, also accompanied by increased p53 mRNA expression. The conclusion was that induction of B-cell differentiation involves the transcription of the p53 gene. This was further substantiated by experiments showing that differentiation of stable clones derived from the 70Z/3 cell line, harboring a p53-promoter-CAT plasmid, induced increased CAT activity. Furthermore, wild type p53 transactivated the promoter control sequences of the kappa light chain gene. Taken together, these results suggest that p53 is involved in B-cell differentiation, a pathway which involves DNA rearrangements that may be accompanied by generation of faulty DNA. The fact that wild type p53 was shown to function as a transcriptional factor, coupled with the notion that it is associated with DNA repair systems, may designate p53 as a control protein in the B-cell differentiation pathway. PMID- 8247533 TI - Ionizing radiation and UV induction of p53 protein by different pathways in ataxia-telangiectasia cells. AB - Cell cycle anomalies have been described in ataxia-telangiectasia cells after exposure to ionizing radiation. A recent report demonstrates that cells from these patients lack the ionizing radiation-induced increase in p53 protein seen in controls. We report here that an ionizing radiation-induced p53 response is reduced and/or delayed in cells from four ataxia-telangiectasia complementation groups. On the other hand, p53 induction is normal in all A-T complementation groups after exposure to UV-B light, an agent to which these cells are not hypersensitive. Specific inhibitors of protein kinase C and serine/threonine phosphatases prevented the radiation induction of p53 protein. Agents that produced double-strand breaks in DNA and/or inhibition of transcription caused an induction of p53 in the absence of radiation in control cells but not in ataxia telangiectasia, but inhibitors of cell cycle progression such as mimosine and aphidicolin led to an increase in p53 in both cell types in the absence of radiation. These results suggest that there is more than one signal transduction pathway responsible for activation of p53, one of which is less efficient in ataxia-telangiectasia cells. PMID- 8247534 TI - v-raf confers CSF-1 independent growth to a macrophage cell line and leads to immediate early gene expression without MAP-kinase activation. AB - The BAC-1.2F5 macrophage cell line depends on CSF-1 for proliferation and survival. Phosphorylation and activation of the RAF-1 kinase are among the early events in CSF-1 signal transduction. To characterize the role of RAF-1 in CSF-1 induced proliferation, we overexpressed oncogenically activated RAF-1, cellular RAF-1 and RAF-1 kinase-defective mutant proteins in BAC-1.2F5 cells. We were unable to establish stable cell lines expressing either kinase-negative or full length RAF-1 proteins, implying that expression of these molecules is not tolerated in BAC-1.2F5 cells. Oncogenically activated RAF-1 induces CSF-1 independent growth in the absence of autocrine growth factor production. Autonomous growth is not associated with dedifferentiation, since v-raf expressing macrophages perform the same immunological functions as control cells. Intriguingly, autonomous growth correlates with the suppression of CSF-1-mediated MAP-Kinase activation and with the low constitutive expression of a number of CSF 1-inducible genes, including fos, jun, ets2, and myc, but also the genes for the inflammatory cytokines TNF alpha and IL-1 beta. Many of these genes have AP-1 binding sites in their promoters, and the v-raf-expressing cells contain constitutive AP-1 binding activity. These data indicate that RAF-1, but not MAP Kinase, is a key component in CSF-1 mitogenic signal transduction, and are consistent with a working hypothesis in which RAF-1 mediates transcriptional activation of genes via AP-1. PMID- 8247535 TI - Timing of SV40 oncogene activation by site-specific recombination determines subsequent tumor progression during murine lens development. AB - We generated mice that carry copies of a dormant transgene encoding the SV40 tumor antigens. The transgenes are specifically targeted to the lens and contain features that render their expression dependent on the action of Cre, a site specific bacteriophage DNA recombinase. Timing of oncogene activation was controlled by making Cre available either prior to, or coincident with, the onset of primary fiber differentiation in the embryonic lens vesicle. Early expression of Cre resulted in oncogene activation in undifferentiated lens epithelial cells that rapidly proliferated inside the lens capsule. By contrast, when Cre accumulation was delayed to coincide with the onset of primary lens fiber differentiation, SV40 oncogenes were activated in cells that had begun to elongate and to accumulate lens-specific crystallins. During subsequent proliferation inside the lens capsule, transformed progeny cells maintained the profile of fiber differentiation that their parent cells had acquired at the time of oncogenic conversion. Developing lens tumors were confined within the capsule of the embryonic lens. However, if the capsule was perforated in an embryonic eye in organ culture, cells rapidly grew out while still maintaining features of differentiation. Our findings show that the differentiated state of the primary target cells is an important parameter of subsequent lens oncogenesis, and that an intact lens capsule can restrict invasive neoplastic growth. PMID- 8247536 TI - Regional localization of Fyn in adult brain; studies with mice in which fyn gene was replaced by lacZ. AB - Mutant mice in which beta-galactosidase gene (lacZ) was inserted into fyn locus were generated by homologous recombination in embryonic stem cells to examine the Fyn expression in the central nervous system. In adult brain, intensive beta galactosidase activity was observed in olfactory bulb, cerebellum and hippocampus of the limbic system; the subcellular distribution of the activity was apparent not only in cell body but also in neural processes, and homozygous mutant mice live-born displayed an anatomical abnormality in the neural cell layer of the hippocampal formation. In spinal cord it was specifically expressed in dorsal horn, and in brain stem it was more characteristic in the sensory pathway, suggesting roles of Fyn in the sensory nervous network. In the white matter area, it was intense at postnatal day 10 but not detectable in adult, suggesting Fyn's role in myelinization. PMID- 8247537 TI - Translocation junctions cluster at the distal short arm of chromosome 1 (1p36.1 2) in human neuroblastoma cells. AB - Human neuroblastoma cells show at high frequency structural changes of the distal short arm of chromosome 1 (1p). The commonly altered region has been identified in previous loss-of-heterozygosity (LOH) studies to involve deletion of 1p36.1-2. These bands are also the site of constitutional alterations in patients with neuroblastoma. In an approach to define the 1p36.1-2 alterations in more detail we here employ four neuroblastoma cell lines to map translocation breaks involving 1p36.1-2 by fluorescence in situ hybridization (FISH). A chromosomal interval flanked by loci DIS96 and DIS98 contained translocation junctions in each of four lines. This analysis identifies in 1p a restricted genomic region as involved in chromosomal rearrangement in different neuroblastomas. The specificity of neuroblastoma translocation junctions at the molecular level implicates this genomic region in tumor development. PMID- 8247538 TI - ER-localization and functional expression of the HBV transactivator MHBst. AB - In two recently reported cases, integrated hepatitis B virus (HBV) DNAs cloned from hepatocellular carcinoma were found to express a transcriptional transactivator from 3'-terminally truncated HBV surface (preS/S) genes. In this study, we characterized the transactivator at the protein level. Expression of a 3'-truncated preS2/S gene in Spodoptera frugiperda (Sf9) insect cells resulted in a C-terminally truncated middle surface protein of 76 amino acids (MHBst76), which was found to be associated with membranes of the endoplasmic reticulum and retained from Golgi processing and secretion. Accordingly, the microsome fraction of MHBst76-expressing Sf9 cells displayed transactivator activity after electric field-mediated transfer into Chang liver cells. In contrast to full-length MHBs, MHBst76 is unglycosylated, and glycosylation is not required for transactivation as shown by mutation of the glycosylation site at asparagine-4. Since highly purified MHBst76 derived from an E. coli expression system also showed transactivator activity, it is concluded that unglycosylated MHBst76 protein is the authentic transactivating factor. As the transactivator protein derives from inactive MHbs by rearrangements of integrated HBV DNA, it may be important for HBV-associated liver carcinogenesis. PMID- 8247539 TI - Characterization of lethal alleles of D-elg, an ets proto-oncogene related gene with multiple functions in Drosophila development. AB - We have used genetic complementation rescue to identify two lethal alleles of D elg, an ets proto-oncogene related gene of Drosophila. Animals that are hemizygous or trans-heterozygous for the alleles die as pharate adults, demonstrating normal gene function is required to complete Drosophila development. Females trans-heterozygous for the 1(3)902 lethal allele and the previously characterized tne female sterile allele produce embryos with abdominal segmentation defects. This finding implicates a role for the D-elg gene in anterior-posterior patterning. The cloning and sequencing of the lethal alleles identified molecular mutations that may result in ELG protein truncation, altered ELG protein interactions, or defective D-elg mRNA splicing. PMID- 8247540 TI - PEA3, Oct 1 and Oct 2 positively regulate the human ETS1 promoter. AB - The promoter of the human ETS1 gene contains binding site motifs for transcription factors Sp1, Ap1, Ap2, ETS, PEA3 and Oct. It has been shown previously that Ap1, Ap2, and ETS are positive regulators for the transcription of the ETS1 gene and that the ETS1 gene is autoregulated. In addition, two regions on the ETS1 promoter that contain negative regulatory sequences have also been identified. In this study, GST/PEA3 and GST/ets1 fusion proteins were used in gel mobility assays to analyse the interaction between these transcription factors with the binding motifs of PEA3 and ETS in the ETS1 promoter. Promoter constructs in which the binding site motifs are mutated were used to examine the effect of mutation on the activities of transcription factors PEA3, ets1 and Oct. Reporter plasmids containing different deletion mutants of the ETS1 promoter were used to examine the effect of different transcription factors (PEA3, Oct 1 and Oct 2) upon the expression of the ETS1 gene. The results of these studies indicate that PEA3 is a strong positive regulator of the ETS1 promoter and that other transcription factors increase the activity of the ETS1 promoter in an additive rather than synergistic fashion. PMID- 8247541 TI - Apoptosis in Burkitt lymphoma cells is driven by c-myc. AB - Chromosomal translocation and subsequent de-regulation of the c-myc proto oncogene are considered to be critical events in the multi-stage evolution of Burkitt lymphoma (BL). It is widely accepted that Myc protein functions as a competence factor for proliferation. However, recent studies indicate that it can also act in some cell types as a regulator of apoptosis. BL cell populations display a high frequency of apoptosis in vivo, a property which is also readily demonstrable in vitro in group I BL cell lines. Such lines are known to retain the cell surface marker characteristics of the parental tumour cells and, in the case of Epstein-Barr virus-positive tumours, their restricted viral protein expression. We have shown previously that apoptosis in a group I BL cell line is inhibited by interferon (IFN)-alpha. Here we show that IFN-alpha-mediated suppression of apoptosis in group I BL cells corresponds temporally with inhibition of Myc protein levels. Furthermore, inhibition of Myc expression following treatment with c-myc anti-sense oligonucleotides markedly enhanced survival of group I BL cells. These results indicate that, whilst c-myc may facilitate cycling of tumour cells in which it is de-regulated, it also stimulates their apoptosis. PMID- 8247542 TI - Transcriptional regulation of the c-erbB-3 gene in human breast carcinoma cell lines. AB - The c-erbB-3 gene is a member of the EGF receptor family of membrane tyrosine kinases. Previous work has shown that the expression of this gene is frequently unregulated in human breast carcinoma-derived cell lines and in tumour biopsy material. As there is no evidence of gene amplification, we decided to examine the control of c-erbB-3 transcription in overexpressing cells. In this paper we describe the cloning of the c-erbB-3 promoter and its functional analysis within mammary-derived cell lines. We show that the promoter is more active in overexpressing cells and this is largely due to the action of the OB2-1 transcription factor which we have recently identified as mediating overexpression of the c-erbB-2 promoter in similar cell lines. PMID- 8247543 TI - A survey of protein tyrosine kinase mRNAs expressed in normal human melanocytes. AB - We have used the reverse transcription-polymerase chain reaction to survey the repertoire of protein tyrosine kinases expressed in cultured normal human melanocytes, a differentiated cell type derived from the neural crest. We identified 25 different tyrosine kinase cDNAs among a total of 608 protein tyrosinase kinase-related cDNAs analyzed. Six encode receptor tyrosine kinases for known ligands, several of which have been implicated in controlling melanocyte proliferation in vitro. Two others encode apparent receptor tyrosine kinases for unknown ligands. Four encode known non-receptor tyrosine kinases and five encode previously identified anonymous protein tyrosine kinases. Of the eight other melanocyte-associated protein tyrosine kinases, most or all appear to be novel. These 25 protein tyrosine kinase genes exhibit distinct patterns of expression in cultured human melanocytes, human erythroleukemia cells, and a variety of normal human tissues. We mapped 16 of the corresponding protein tyrosine kinase genes to specific human chromosomes, identifying a total of 19 human genetic loci, some of which may constitute candidate genes for genetic disorders of mammalian development. PMID- 8247544 TI - Wild type p53 can mediate sequence-specific transactivation of an internal promoter within the mdm2 gene. AB - The p53 tumor suppressor gene product can complex with polypeptides encoded by the mdm2 putative protoncogene. In addition, mdm2 mRNA levels have been shown to increase following the activation of wild type (wt) p53. To determine the basis for the effect of wt p53 on mdm2 mRNA, we studied the interaction of the mdm2 gene with p53. We report that wt p53 can bind sequence-specifically to a DNA region residing downstream to exon 1 of the mdm2 gene. This is correlated with a pronounced p53-dependent transcriptional activation. Efficient p53-dependent transactivation can be obtained with an mdm2 genomic DNA fragment lacking the putative mdm2 promoter. These findings suggest that p53 can induce transcription from an internal promoter located within the mdm2 gene. These findings raise the possibility that, in addition to increasing the overall levels of mdm2 mRNA, wt p53 may also modulate the repertoire of mdm2 transcripts present within the cell. PMID- 8247545 TI - Cell-type- and promoter-dependent ts phenotype of p53 Val135. AB - The p53 mutant Val135 is widely considered to have a wild-type (wt) phenotype at 32.5 degrees C, but not at 37 degrees C. The ability of wt murine p53 and its Val135 mutant to modulate transcription from the muscle-specific creatine kinase promoter (-3.3 kb pMCK), from a reporter construct containing two copies of the p53-binding DNA element from within MCK (p50-2), and from the interleukin-6 (IL 6) promoter (pIC225) was evaluated in transient transfection experiments in CV1 and HeLa cells. In CV1 cells, wt p53 was confirmed to activate the pMCK and p50-2 reporters, but to repress the IL-6 promoter. However, although in these cells p53 Val135 had the expected wt-like phenotype with respect to activation of the p50-2 reporter at 32.5 degrees C (32.5 degrees C > 37 degrees C), this mutant had little effect on expression from pMCK at either temperature, and activated rather than repressed the IL-6 promoter at 32.5 degrees C. In HeLa cells, although wt p53 activated p50-2 but repressed the MCK and IL-6 promoters, p53 Val135 activated all three reporters. Unexpectedly, in these cells the upregulation of p50-2 and pIC225 was basically temperature-independent, and that of pMCK was inversely ts (37 degrees C > 32.5 degrees C). The novel ts properties of p53 Val135 show that this mutant is not always wt-like at 32.5 degrees C but exhibits strong cell-type and promoter-dependent differences in its ts phenotype for transcriptional modulation. PMID- 8247546 TI - Binding of cellular proteins to a conformational domain of tumor suppressor protein p53. AB - The genes regulated by p53, as well as the factors modulating its function, need to be identified before the mechanism of action of p53 in control of cell growth can be adequately understood. Binding of the SV40 large T-antigen protein to an evolutionally conserved (conformational) domain of p53 inhibits p53's DNA-binding and transcription activation activities. Cellular proteins might also bind to this same region of p53 to regulate its function. A hybrid protein composed of protein A fused to the conformational domain (amino acids 115-295) of p53 was expressed in Escherichia coli and used as an affinity probe for binding proteins in detergent lysates of non-small cell lung carcinoma (NSCLC) cells. The wild type p53 hybrid protein associated with several major proteins of molecular weights 45 K, 56 K, and 70 K, as well as other minor species ranging in molecular weight from 30 K to 90 K. These proteins bound specifically to the p53 sequence of the hybrid protein. Protein A did not associate with these proteins and the two p53 hybrid proteins containing missense mutations at codons 273 and 175 exhibited a 40-80% weaker association. In addition, T antigen competed with the cellular proteins for binding to the conformational domain. The conditions of cell growth had a profound effect on the expression of the p53 binding proteins. Considerably more p53 binding proteins were expressed in actively growing cells than in cultures maintained under conditions for slow growth. Quantitative differences in expression of p53-binding proteins were observed among different NSCLC cell lines. The expression of p53-binding proteins was not restricted to NSCLC cell lines; detergent extracts of an osteosarcoma cell line yielded similar p53-binding proteins. PMID- 8247547 TI - Wild-type p53-triggered apoptosis is inhibited by bcl-2 in a v-myc-induced T-cell lymphoma line. AB - Using a temperature sensitive p53 construct (ts p53), we have earlier shown that expression of wild-type (wt) p53 triggers apoptosis in a v-myc-induced T-cell lymphoma line that lacks endogenous p53, and in a Burkitt lymphoma line that carries mutant p53. We have suggested that apoptosis is elicited by the contradictory signals emanating from the constitutively activated myc gene and the growth arresting signal of wt p53 (Ramqvist et al., 1993; Wang et al., 1993). Work in other laboratories has shown that constitutive c-myc expression can induce apoptosis when cell proliferation is inhibited due to the lack of growth stimulating factors. Expression of bcl-2 could inhibit apoptosis. In order to test whether p53-induced apoptosis can be prevented by bcl-2, we have introduced a retrovirally driven bcl-2 construct into our v-myc-induced murine T-cell lymphoma line, previously transfected with ts p53. About 90% of the parental ts p53 transfected cells died of apoptosis within 3 days after induction of wt p53 expression at 32 degrees C. Two clones of ts p53/bcl-2 double transfectants that expressed high levels of bcl-2 from the introduced construct were completely protected from apoptosis, following transfer of the cells to 32 degrees C. One clone that expressed the exogenous bcl-2 only at a low level was partially protected from wt p53-induced apoptosis. Clones of the parental ts p53 carrying cells transfected with the puromycin resistance gene vector, without the bcl-2 gene underwent 90% apoptosis. These results suggest that bcl-2 may prevent apoptosis in cells simultaneously exposed to the proliferation-stimulating effect of activated myc and the growth arresting signal of wt p53. PMID- 8247548 TI - Structure, expression and chromosomal mapping of TKT from man and mouse: a new subclass of receptor tyrosine kinases with a factor VIII-like domain. AB - Using a polymerase chain reaction-mediated approach we have characterized cDNAs from human and mouse origin representing a novel type of receptor protein tyrosine kinase (RTK). The deduced amino acid sequence (855 amino acids) of the longest open reading frame has a unique extracellular region encompassing a factor VIII-like domain, not previously described for RTKs. The most closely related RTKs are members of the neurotrophin receptors (TRK), which showed 47-49% homology with the kinase domain of the new RTK. Therefore, the new gene has been called TKT (Tyrosine-Kinase related to TRK). TKT orthologs from man and mouse were 98% similar. In both species a major transcript of 10 kb was found to be expressed at high levels in heart and lung. Low levels of this mRNA-species were detected in human brain, placenta, liver, skeletal muscle, kidney and in mouse brain and testis. Analysing human/mouse somatic cell hybrids we demonstrated that TKT segregates with human chromosome 1. PMID- 8247549 TI - New activated RAS2 mutations identified in Saccharomyces cerevisiae. AB - Activating mutations in RAS proto-oncogenes encode proteins with greater GTP binding. Such mutant proteins are responsible for many human cancers. Six new amino acids were discovered that can yield an activated Saccharomyces cerevisiae RAS2 protein when they are altered. These new RAS2 alleles were found among a collection of 35 random mutations that exhibit a dominant reduction of glycogen accumulation. The RAS2-P41S and RAS2-E99K alleles encode proteins that have lost responsiveness to GTPase activating proteins. They affect amino acids in loop 2 and helix 3 respectively and illustrate that GTPase activating proteins recognize a larger portion of the RAS structure than previously realized. RAS2 mutations E130K, S153F, A154T, and A157S alter amino acids proximal to the guanine binding site and probably influence nucleotide binding either directly or indirectly. PMID- 8247550 TI - Overexpression of cyclin D1 in rat fibroblasts causes abnormalities in growth control, cell cycle progression and gene expression. AB - Cyclin D1, a putative G1 cyclin, has been implicated in cell cycle control. The human cyclin D1 gene is located on chromosome 11q13 where DNA rearrangement and amplification have been detected in several types of human cancer. Previous studies demonstrated that the cyclin D1 gene is not only rearranged or amplified but also overexpressed in some of these human tumors and tumor-derived cell lines. To further address the roles of cyclin D1 in cell cycle control and tumorigenesis, we have stably overexpressed the human cyclin D1 cDNA in Rat6 embryo fibroblasts by using retrovirus mediated transduction. The cyclin D1 protein was overproduced about 10-fold and was localized predominately in the nucleus. Cyclin D1 overexpressing cells displayed a decrease in the duration of the G1 phase, decreased cell size, and induced tumors when injected into athymic (nude) mice. In addition, overexpression of cyclin D1 in Rat6 cells perturbed the expression of several cellular growth-related genes including c-myc, c-jun, and cyclin A, but not cyclin D3. Taken together, these results indicate that deregulated expression of the cyclin D1 gene can cause disturbances in cell cycle control and gene expression and also enhance tumorigenesis. PMID- 8247551 TI - Transcriptional activation domains of elk-1, delta elk-1 and SAP-1 proteins. AB - elk-1, an ets related gene codes for a sequence specific DNA binding transcriptional activator which in association with serum response factor (SRF) forms a ternary complex at the c-fos serum response element (SRE). Recently the C terminal region of both elk-1 and delta elk-1 proteins was shown to undergo phosphorylation by MAP kinases and function as an activator of MAP kinases. Here we show that delta elk-1 and two other elk-1 related proteins SAP-1a and SAP-1b, like elk-1, can function as transcriptional activators. In this report we have localized the transcriptional activation domain of the SAP-1 proteins (STA) to a large portion of the carboxy terminal region and have identified two autonomous transcriptional activation domains in the elk-1 protein, one at the amino (ETA-1) and the other at the carboxy terminal region (ETA-2). delta elk-1 protein contains only the ETA-2 domain indicating differential usage of activation domains as a result of alternative splicing. We can speculate that the ETA-1 domain can function in vivo independent of ETA-2, but the ETA-2 domain can function either in the absence of ETA-1 (as seen in delta elk-1) or in the presence of accessory proteins like SRF. The role of SRF in the activation of the ternary complex might be to bind to the ETA-1 domain, somehow conceal it's activation domain and in the process unmask the ETA-2 domain (for phosphorylation by MAP kinases) and activate transcription. The ETA-1 domain may be functioning as a negative regulatory transcriptional activation domain for ETA-2. These observations suggest that the elk-1 family of proteins may not only regulate fos and MAP kinases but also other elk-1 target genes that are essential for cellular growth control. PMID- 8247552 TI - The retinoblastoma-related gene, RB2, maps to human chromosome 16q12 and rat chromosome 19. AB - A retinoblastoma-related human gene, referred to as RB2, has been cloned based on sequence homology of the E1A-binding domain of the retinoblastoma gene. Structural homology with the retinoblastoma gene suggests a possible function of RB2 as a tumor suppressor gene. In this study, we have mapped this gene to human chromosome 16q12.2 and rat chromosome 19, using fluorescence in situ hybridization and somatic hybrid cell analysis, respectively. Based on known syntenic relationships among human, rat and mouse, the data suggest that the mouse homolog resides on chromosome 8. Deletions of chromosome 16q have been found in several human neoplasias (including breast, ovarian, hepatic, and prostatic cancers) which is in support of an involvement of RB2 in human cancer as a tumor suppressor gene. PMID- 8247553 TI - Physicians and the environment: an important alliance. PMID- 8247554 TI - An outline for action: how physicians might become involved in the environmental debate. PMID- 8247555 TI - The American Lung Association: a partnership of science and advocacy in defense of the environment. PMID- 8247556 TI - Impact of the environment on blood and blood-forming tissues. PMID- 8247557 TI - Public health and responsible environmental stewardship: some opportunities for leadership by physicians. PMID- 8247558 TI - A medical dilemma: quality care vs. cost reduction. PMID- 8247559 TI - Orbital complications in functional endoscopic sinus surgery. AB - Endoscopic sinus surgery can result in both minor and major complications. Among these, orbital complications--including retroorbital hematoma--are among the most feared. Injuries can be direct or indirect from pulling on diseased structures. A retrospective chart review of 616 endoscopic sinus procedures revealed eight orbital complications in seven patients. These included two medial rectus injuries, five orbital hemorrhages, and one nasolacrimal duct injury. Predisposing factors may include hypertension, lamina papyracia dehiscences, extensive polypoid disease, previous surgery, inability to visualize the maxillary ostia, violent coughing or sneezing, and chronic steroid use. Suggested management in the literature includes lateral canthotomy, steroids, and mannitol with ophthalmologic consultation. Opening of the wound by means of an external ethmoidectomy incision has also been suggested. We suggest that adding orbital decompression by means of multiple incisions into the periorbita should be added for fully effective relief. A "management" tree of decision parameters relevant to orbital complications is presented. PMID- 8247560 TI - Ingestion of caustic cosmetic products. AB - Twelve children over a 6-year period (1985 to 1991) underwent aerodigestive tract endoscopy after ingestion of lye-containing cosmetic products. All children were less than 2 years of age and had facial and/or oral cavity burns. Eleven of these patients ingested alkaline hair products such as "activators" or "relaxers." Endoscopy revealed pharyngeal burns in five patients but no laryngeal or esophageal burns in any patient. Erythema of the esophageal mucosa was seen in one patient. One patient ingested a solution used in fingernail decorating. This patient sustained extensive superficial burns of the pharynx, larynx, and esophagus and required intubation and admission to the intensive care unit. No long-term sequelae developed. Cosmetic products containing caustic chemicals represent a significant health risk to small children, especially because child proof packaging is not standard for these products. PMID- 8247561 TI - A practical methodology to analyze facial deformities. AB - Despite much information about various methods to analyze the face, a practical stepwise methodology has not been firmly established in otolaryngology teaching programs. A general guideline for facial analysis would be helpful to individuals not totally familiar with facial analysis for assessing facial deformities resulting from to traumatic, neoplastic, or developmental causes. This article attempts to distill and integrate previously established guidelines for facial analysis into eight major steps to provide a preliminary and practical methodology for initial facial assessment. PMID- 8247562 TI - The role of KTP laser in revision stapedectomy. AB - In recent years, the safety and efficacy of revision stapedectomy has come under scrutiny. Experienced surgeons report that the results of such surgery are often worse than the results after primary surgery and that the risks of sensorineural hearing loss, tinnitus, and vertigo are increased. With the addition of laser technology to revision stapes surgery, the procedure to open the neomembrane over the oval window and gain access to the inner ear can now be performed safely. This allows positive identification of the oval window and assures placement of the prosthesis through the fenestra rather than on an intermediate segment of scar or bone in the region of the footplate. Our studies have shown the laser to be an important tool that enhances the safety and efficacy of revision stapedectomy. PMID- 8247563 TI - The biochemistry of neutralization. AB - It has been shown that injections of low doses of common environmental antigens or irritants cause a reduced sensitivity during a subsequent exposure to the same antigen and irritant. On the basis of these findings, the following study was performed: 20 allergic patients who manifested allergic symptoms were observed during skin end-point titration testing, which is described in the manuscript. During the testing procedure, 60% of the patients reported a complete relief of symptoms. It is postulated that a cytoprotective dose was given during the testing procedure that caused the symptoms to be relieved. The second part of the study was a double-blind placebo control study in which 40 patients received the end-point dose and 10 patients received placebo. All patients entered the study with allergic symptoms. None of the placebo patients reported any relief of symptoms when given their injection. Of patients who received the active ingredient, 67.5% reported relief of symptoms within 5 to 10 minutes after the subcutaneous administration of the active ingredient. With these findings, it is postulated that this low dose of active ingredient caused the production of prostaglandin E intracellularly, which causes an increase in cyclic AMP and a decrease in cyclic GMP, which results in the resolution of symptoms. PMID- 8247564 TI - Petrous jugular malposition (diverticulum). AB - Jugular bulb anatomy is variable. A "high-riding" bulb extending into the tympanic cavity is a well-described anomaly. Petrous jugular malposition (diverticulum) (PJMD), however, is rare. The relationship between PJMD and clinical symptoms is questionable because the differentiation between PJMD as an anatomic variant and pathologic process is unproved. A literature review reveals 14 previously documented cases. We report an additional four cases. Diagnostic and management dilemmas are discussed, with the importance of high-resolution CT stressed. PMID- 8247565 TI - Neurotologic manifestations of Chiari 1 malformation. AB - It is important for otolaryngologists to recognize Chiari malformations as part of the differential diagnosis of balance disorders because patients may initially be seen with symptoms referable to the vestibular system, including ataxia, nystagmus, or vertigo. The objective of this paper is to review the signs, symptoms, and vestibular test findings of a series of patients with Chiari 1 malformation. Six patients were identified by retrospective chart review with a diagnosis of Chiari malformation. Each patient had a complete otoneurologic examination and vestibular function testing. The results indicated that patients fell into two different vestibular test result profiles. First, patients with advanced symptoms demonstrated oculomotor dysfunction, central vestibular nystagmus, abnormal vestibular visual interaction, and abnormal tilt suppression of postrotatory nystagmus. On the other hand, a number of patients were identified with incidentally noted Chiari malformation on magnetic resonance imaging scan who had a vestibular test profile consistent with peripheral vestibulopathy without signs and symptoms of central nervous system dysfunction. Guidelines are provided to help determine the extent of the group of symptoms attributable to an incidentally discovered Chiari malformation. PMID- 8247566 TI - Sources of error in use of beta-2 transferrin analysis for diagnosing perilymphatic and cerebral spinal fluid leaks. AB - Beta-2 transferrin is a protein found in cerebral spinal fluid and inner ear perilymph, but not in blood, nasal, or ear secretions. The clinical use of this test has been previously demonstrated, but sources of test error have not been addressed. The purpose of this study was to evaluate sources of error related to this test in order to improve its clinical use. We reviewed the specimens submitted for beta-2 analysis over the first 12 months of test availability at our institution to identify potential factors leading to test error. Sources of error were categorized into the following groups: sample collection, delivery, and extraction factors; assay factors; physician-related factors; and patient related factors. The test for beta-2 transferrin is a valuable diagnostic tool for the management of difficult clinical problems, provided the physician is aware of potential factors that can lead to test error and clinical mismanagement. PMID- 8247567 TI - Atypical presentation of acoustic neuroma. AB - Unilateral progressive sensorineural hearing loss, tinnitus, and unsteadiness are the usual initial symptoms of acoustic neuroma. Of the last 100 consecutive cases of acoustic neuroma detected at our Centre, 14 had atypical symptoms, Five patients manifested sudden hearing loss; one of these had complete recovery. Three patients reported long-standing unilateral hearing loss, ranging from 10 to 20 years. Six patients had normal hearing, one of whom was diagnosed incidently when the investigations were performed for contralateral glomus tumor. A second patient, a young woman, experienced weakness of lower limbs. The remaining four patients had only subjective symptoms of hearing loss or tinnitus. Acoustic tumors could have been overlooked easily in these patients. It is important to have a high index of suspicion in all cases of sudden hearing loss, asymmetric sensorineural loss of any duration, subjective sensation of hearing loss, and tinnitus. It is mandatory to investigate these cases with auditory brainstem responses, any abnormality of which makes it necessary to perform magnetic resonance imaging with gadolinium. PMID- 8247568 TI - Middle ear adenoma and adenocarcinoma. AB - Adenomas and adenocarcinomas originating from the middle ear are few in number and have met with significant controversy in the literature. Our experience with 11 previously described patients is updated and five new cases are added. These tumors were seen in persons whose ages ranged from 7 to 77 years. The most common initial symptom was decreased hearing (11 cases), followed by otorrhea (4 cases), and otalgia (4 cases). At the time of diagnosis, facial nerve weakness was present in several patients (seven), and this was a poor prognostic sign. These glandular tumors engender controversy regarding histologic origin and the pathologic demarcation between adenoma and adenocarcinoma. In this review, we show that otorrhea, bone erosion, facial nerve involvement, and aggressive clinical behavior can be the result of lesions that appear microscopically benign. Furthermore, we present evidence of the similarity of these glandular lesions to paragangliomas and carcinoid tumors. Finally, pathologic data are presented to develop the previously unreported theory that the origin for this group of tumors is paraganglionic tissue. PMID- 8247569 TI - Extramedullary plasmacytomas of the head and neck. AB - Extramedullary plasmacytomas are solitary tumors consisting of neoplastic plasma cell proliferations that occur in locations other than bone. On initial presentation they must be differentiated from multiple myeloma. This may prove to be difficult because a varying percentage may be associated at a later date with the development of multiple myeloma. Solitary extramedullary plasmacytomas represent up to 4% of nonepithelial lesions of the upper respiratory tract. From 1970 to 1990, at West Virginia University Hospitals, seven patients with solitary extramedullary plasmacytoma were identified. In four of these patients the tumor was located in the head and neck, with one tumor located in each of the following sites: temporoparietal scalp, maxillary sinus, nasopharynx, and cervical region. One patient had extensive destruction of the temporal bone, with extension intracranially to the middle cranial fossa. No patient had multiple myeloma, nor did any develop. Diagnosis was based on a combination of histology along with special immunoperoxidase staining for Ig lambda and kappa light chains. This will be demonstrated and discussed. Treatment consisted of radiotherapy in three cases, with doses ranging from 3175 to 6000 rad. One patient, treated with surgical excision, experienced a relapse at a distant site 6 years later. All patients have maintained local control and have been followed for a minimum of 1 1/2 years, with an average of 3 years. We describe our experience with these tumors and present a pertinent review of the literature. While these tumors may present as aggressive locally destructive lesions, their management should be as organ-sparing as possible because excellent control can be achieved in the majority of cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247570 TI - Pediatric sinusitis and subperiosteal orbital abscess formation: diagnosis and treatment. AB - Twenty-two children with subperiosteal orbital abscesses were treated at St. Louis Children's Hospital between 1983 and 1992. Eighteen patients were otherwise in good health; four patients had cystic fibrosis, mucoceles, or were immunocompromised. All patients were treated with intravenous antibiotics and abscess drainage. CT scans were obtained preoperatively in each case. Ten patients were treated with endoscopic ethmoidectomy and abscess drainage, and 11 were treated with external ethmoidectomy and abscess drainage. One child was initially treated with abscess drainage and an elective endoscopic ethmoidectomy was performed later. There were no cases of permanent visual loss or neurologic sequelae. Culture results were positive in 14 cases and mixed infections were common. Complications included recurrent abscess, cerebritis, and empyema. We recommend combined medical and surgical treatment for all children with subperiosteal orbital abscess. We feel that endoscopic ethmoidectomy and abscess drainage offers some advantages over external ethmoidectomy and abscess drainage. PMID- 8247571 TI - An education model for otitis media care field-tested in Latin America. AB - The World Health Organization has designated the teaching of otitis media management skills a "priority" status. Effective treatment of ear disease requires that the physician be both informationally educated as well as physically trained to use otoscopy. Little is known about how well this education can be provided in a short time and in a foreign country. To more objectively assess teaching effect, results of an education session for rural Mexican pediatric primary-care providers who were given an intensive otitis media lecture and otoscopy skills workshop in 1990 were evaluated. To test immediate cognitive impact, an anonymous written examination was given both before and after the teaching session. Average test scores after the educational sessions improved 24% (p < 0.001) over baseline scores before the sessions. To evaluate long-term impact on clinical practice, a follow-up telephone survey 2 years later was conducted. The use of an otoscope to diagnose otitis media had increased from 40% to 93% of respondents. We conclude that pediatric primary-care providers in rural Mexico possess a baseline level of knowledge about otitis media that can be significantly enhanced with one educational session. Further, this teaching effort produces an impact on practice pattern that lasts at least 2 years. PMID- 8247572 TI - Effect of changes in mass on middle ear function. AB - Vibrating systems such as the middle ear are affected by changes in mass. After disease or ear surgery, significant changes in mass may contribute positively or negatively to the postoperative hearing threshold. This article describes experiments in 15 human temporal bones of the addition or reduction of mass on the middle ear transfer function. Measurement of stapes and umbo vibration was performed using a Laser Doppler Vibrometer before and after the addition of different masses at several sites on the tympanic membrane (TM) and ossicular chain. The input was 61 pure tones swept from 147 to 19433 Hz at 80 dB SPL. The addition of mass onto the TM produced varying detrimental effects on sound transmission, depending on the location and amount of mass. The insertion of ventilation tubes, weighing 12 to 17 mg each, produced losses at 1.5 to 5.0 kHz compared with tympanotomy alone. Addition of mass to the umbo and malleus head produced a loss at mid and high frequencies, whereas addition of mass on the incus long process and stapes also produced a high-frequency decrease in stapes displacement. Reduction of TM mass by removal of the epithelium produced an increase, especially at 2.0 to 4.0 kHz. PMID- 8247573 TI - TGF-alpha protein and receptor localization in laryngotracheal tissue. AB - Both normal cell turnover and healing of laryngeal and tracheal injuries involve cell migration and mitosis. The proteins that regulate normal cell turnover and wound healing in the larynx and trachea have not been established. It is possible that peptide growth factors, such as transforming growth factor-alpha (TGF alpha) acting through its receptor (EGF/TGF alpha-R), participate in the regulation of these processes. To investigate this hypothesis, we analyzed laryngotracheal cells for TGF alpha protein and receptor in normal and postwounding conditions. TGF alpha protein was detected by immunohistochemical analysis in normal ferret laryngeal and tracheal mucosa. Specific binding to the EGF/TGF alpha receptor in membrane homogenates of ferret larynx and trachea reached saturation after 60 minutes at 37 degrees C, and was effectively displaced by unlabeled epidermal growth factor (EGF) or TGF alpha, but not by unlabeled insulin, angiotensin II, or basic fibroblast growth factor. Scatchard analysis of the specific binding indicated the presence of high-affinity (Kd = 117 pmol) and low-affinity (Kd = 40 nmol) binding sites. The maximum number of available binding sites was 73 fmol/mg protein. Localization of the EGF/TGF alpha receptor by autoradiographic analysis of 125I-EGF binding to sections of normal ferret larynx and trachea revealed EGF/TGF alpha receptors throughout the epithelium, with the highest grain density in the basal layers. Quantitative analysis of autoradiographic grain density between normal, intubated, and extubated animals revealed no significant differences. The presence of TGF alpha protein and its receptor in normal and wounded larynx and trachea supports the hypothesis that these proteins are involved in regulating physiologic responses of laryngotracheal cells. PMID- 8247574 TI - Assessment of survival and microscopic changes in porcine skin flaps undergoing immediate intraoperative tissue expansion. AB - The technique of rapid intraoperative tissue expansion has been used with increasing frequency in the clinical setting over the last several years. This technique takes advantage of the skin's ability to immediately stretch and increase in surface area when expanded under a constant load. Sixteen random pattern, rapidly expanded skin flaps on 10 domestic male pigs were studied to assess the predictive value of the fluorescein test for flap viability after rapid intraoperative tissue expansion. Partial fluorescence was found to be a more accurate predictor of flap survival in the experimental rapidly expanded flaps when compared to full fluorescence. Partial fluorescence was found to under predict flap survival by 0.3 to 0.5 cm, whereas full fluorescence was found to under-predict flap survival by 2.5 cm. Additionally, histologic and ultrastructural changes were examined in rapidly expanded skin from the hip region in three pigs. The only microscopic change noted between control and experimental flaps was dilated capillaries in the dermis of expanded skin, which was noted by electron microscopy. Collagen and elastic tissue changes were not demonstrated in rapidly expanded pig skin by electron microscopy, direct immunofluorescence, collagen, and elastic tissue stains. PMID- 8247575 TI - Rhinoscleroma occurring with airway obstruction. PMID- 8247576 TI - Upper airway distress in crack-cocaine users. PMID- 8247577 TI - Second branchial cleft anomalies presenting as pharyngeal masses. PMID- 8247578 TI - Anaplastic carcinoma arising in a thyroglossal duct tract. PMID- 8247579 TI - Ganglion of the temporomandibular joint presentation as a parotid mass. PMID- 8247580 TI - An elongated uvula producing chronic cough. PMID- 8247581 TI - Cholesterol granulomas of the maxillary sinus presenting with nasal obstruction. PMID- 8247582 TI - An immediate custom tracheostomy tube for the thick neck. PMID- 8247583 TI - Surgical pathology specimen orientation markers. PMID- 8247584 TI - Tuberculous laryngitis. PMID- 8247585 TI - Color balance of photographic images. PMID- 8247586 TI - Myth of spontaneous perilymph fistula. PMID- 8247587 TI - Myth of spontaneous perilymph fistula. PMID- 8247588 TI - High ear-piercing. PMID- 8247589 TI - Potency of frozen/thawed botulinum toxin type A in the treatment of torsion dystonia. PMID- 8247590 TI - Tinnitus aereum as an effect of increased tension in the lateral pterygoid muscle. PMID- 8247591 TI - [Sports and physical training in childhood--general principles]. AB - Whether functional performance of children in response to exercise adapts in a similar way to that of adults and exactly what the influence of growth and maturation is in determining a child's performance has not been completely resolved. Regarding to respiratory factors the younger the child the higher the pulmonary ventilation per liter of oxygen consumption. Girls tend to have a higher ventilatory volume in exercise than boys and after puberty it is significantly lower in boys than in girls. Maximal oxygen consumption increases with age in both sexes as a consequence of increased size but aerobic capacity in children is more related to size and body composition than age. Regarding to cardiovascular factors aerobic capacity is largely determined by maximum cardiac output which itself is dependent upon stroke volume and rate of heart beat, because young children have smaller hearts, they have higher heart rates to compensate for the smaller stroke volume compared with adults. Children have higher maximum heart rates than adults, ranging from approximately 195-220 beats/minute. After maturity there is a decrease in heart rate with age of approximately 0.8 beats/min/year of age. Because of the larger hearts greater stroke volumes and an increased blood volume after puberty males have a higher resting systolic blood pressure than females.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247592 TI - [Past, present and future perspectives of the C. E. S. P. (Confederation of European Specialists in Pediatrics)]. PMID- 8247593 TI - [Child abuse. Problems of definition and initial intervention]. PMID- 8247594 TI - Childhood obesity treatment: double blinded trial on dietary fibres (glucomannan) versus placebo. AB - Dietary fibres are frequently used for the treatment of paediatric obesity. The aim of this clinical trial is to evaluate the efficacy of glucomannan in the child obesity management. This experimental design was double blinded with a block randomisation, alpha = 0.05, beta = 0.2 and delta = 50%. The study involved 60 children under 15 years of age (mean age 11.2 years, mean overweight 46%), 30 of them under glucomannan treatment (1 g twice a day for two months) and 30 under placebo and the same schedule. The drug and the placebo were indistinguishable both for the family and the physician. During the two months study period the children followed a normocaloric diet evaluated every two weeks by a dietetic record book. At the beginning of the study the drug and the placebo groups were comparable in regards to anthropometric data. At the end, the mean overweight of the drug group was decreased from 49.5% to 41% and that of the placebo group from 43.9% to 41.7%. Both decreases were significant (p < 0.01), but no significant difference was observed between the drug and the placebo groups. The only significant difference concerned the lipid metabolism. The children under glucomannan treatment manifested a significant decrease of alpha-lipoprotein and an increase of pre-beta-lipoprotein and triglycerides; the children under placebo manifested only a decrease of triglycerides and apo beta-lipoprotein. We suggest that this metabolic alteration may derive from a primary decrease of alpha lipoprotein, most likely because of an inadequate water intake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247595 TI - [An adolescent patient with functional cardiovascular syndrome--treatment with short-term psychotherapy]. AB - This paper presents a case of 14 year old boy with psychogenic heart complaints after the divorce of his parents. Additionally we diagnosed a depressive and dependent personality and neurotic defence mechanisms presenting as somatisation. Psychotherapy was suggestive and supportive and involved imaginative and creative elements as utilized in day dream techniques. This proved to be a sufficient short time therapy and resulted in rapid improvement of the distressing heart pain. The boy was helped to reorganize his life and enabled to adapt sufficiently. This case shows a positive outcome of a multimodal, symptom focused, short time psychotherapy as being sufficient in the treatment of a child with somatisation disorder, due to the high flexibility of the developing personality. PMID- 8247596 TI - [Experience of parents in home monitoring of apnea for prevention of sudden infant death (SIDS)]. AB - 89 of 119 parents of infants with apnea home monitoring answered to a standard questionnaire concerning different aspects of home monitoring. In almost all cases the home monitoring was finished at the begin of the study, lasting from six months to 12 months. The indications for an apnea monitor were: apparent life threatening event (n = 8), SIDS-sibling (n = 24), sleep apnea syndrome (n = 40) and parents pressing request (n = 17). 90% of parents felt to have got enough information about the function of the apnea monitor (mostly Graseby MR-10 monitor) and what they have to do when there is an apnea alarm. 6% of parents changed the monitor because of technical problems. Most of the alarms were registrated by almost all parents between 24 h and 6 h. 35% of parents stimulated their infants in case of an apnea alarm gently to vigorously, one infant had a mouth to mouth resuscitation. Questions concerning the psychological and social impact of home monitoring on parents demonstrated that there was some stress like anxiety, sleep disturbances mainly in the mothers. We conclude from parents experience with apnea home monitoring the need for an adequate medical, technical and psychological support. PMID- 8247597 TI - Pathology of cervical intervertebral disc injuries. AB - Autopsy was performed on 31 patients with traumatic spinal cord injuries who were treated and expired at the Chugoku Rosai Hospital between 1957 and 1987. These patients could be divided into four groups: intervertebral disc injuries (7 patients); hyperextension injuries (9 patients); fracture and fracture dislocation of the cervical spine (11 patients); and fracture-dislocation of the thoracic spine (4 patients). The sex, age, cause of injury, bone injuries seen on roentgenograms, the level and grade of neurological deficits, the survival time, and the findings of the postmortem studies of the 7 patients with intervertebral disc injuries are presented in Table I. This paper illustrates and discusses these 7 cases. PMID- 8247598 TI - Prediction of pulmonary function in wheelchair users. AB - As a result of the improved medical treatment of infectious diseases, these formerly leading causes of mortality in the United States have been supplanted in rank by chronic events as the major causes of death. The major causes include coronary artery disease, respiratory disease, and cancer. Static and dynamic pulmonary functions in the apparently healthy wheelchair user population were measured and then modelled using stepwise regression. One hundred and nine wheelchair users (97 males, 12 females) with paraplegia (n = 77) or quadriplegia (n = 32) gave informed consent and participated in this study. Subjects ranged from being Olympic caliber wheelchair marathon racers to those who live sedentary life styles. Subjects performed three slow vital capacity (SVC) tests, three forced vital capacity (FVC) tests, and three maximal voluntary ventilation (MVV) tests while seated in their standard wheelchair. The order of the pulmonary function tests was randomized. Subjects also completed two functional residual capacity (FRC) measurements. Analysis of variance revealed significant differences in several pulmonary functions based on gender (FEVC, p = 0.0001, FEV1, p = 0.0001, FEVC 25-75%, p = 0.005, PEF, p = 0.002, FIVC, p = 0.002, RV, p = 0.0001, MVV, p = 0.0001, SVC, p = 0.001). The women's unforced prediction equations using age, height, weight did yield some significant correlations with predictions based upon ambulatory subjects. The men's unforced adapted prediction equations did reveal significant correlations with the validation group for FEVC (r = 0.66, p = 0.007), FEV1 (r = 0.62, p = 0.015), PIF (r = 0.95, p = 0.015), MVV (r = 0.57, p = 0.067), SVC (r = 0.69, p = 0.019), and RV (r = 0.67, p = 0.009). Pulmonary function in male wheelchair users should be predicted using equations which incorporate years with disability and level of impairment. Additional study is required to make any recommendations regarding women wheelchair users. However, study of pulmonary function in women should be made a priority. Pulmonary function is affected by the extent of physical impairment, and tends to decline as years with disability increase. PMID- 8247599 TI - Risk of cardiac dysrhythmias in chronic spinal cord injury patients. AB - The incidence of cardiac dysrhythmias is characterized in a group of 47 chronic SCI patients (greater than 30 days post injury; range 35-3605 days) with SCI lesions ranging from C4 to L3 who were referred for evaluation of a potential cardiac dysrhythmia by 24 hour Holter monitoring. Patients were grouped according to level of injury as paraplegic (T1 and below), n = 22, or quadriplegic (C3-C8), n = 25. No evidence of life threatening dysrhythmias was noted in either group, and no differences were evidenced in the incidence of cardiac dysrhythmias between the groups. Premature ventricular contractions (PVCs) were noted in 4/25 quadriplegics, and in 1/22 paraplegics, and right bundle branch block (RBBB) was noted in 1/25 quadriplegics and 1/22 paraplegics. A two-year chart review follow up revealed that no clinically significant cardiac dysrhythmic events occurred in these patients. These findings suggest that the risk of cardiac dysrhythmia is attenuated following the acute phases of SCI and represents a diminished concern to the clinician. PMID- 8247600 TI - Trophic effects on testes in paraplegics. AB - Testicular biopsies and hormone profiles were obtained from 23 paraplegic patients who had sustained a complete spinal cord section. The hormone profiles were normal, but patients with a spinal lesion including the T10-L2 metameres showed a particular pattern of germinal cell abnormalities. The atrophy is multifactorial, but may well include destruction of the sympathetic innervation of the testis by the lesion. PMID- 8247601 TI - Prediction of maximal aerobic power from a submaximal exercise test performed by paraplegics on a wheelchair ergometer. AB - The aim of the present study was to verify the basic principles underlying the prediction of VO2 peak from a submaximal exercise test performed by paraplegics on a wheelchair ergometer and thus to propose regression equations of VO2 peak prediction. Forty-six paraplegic subjects (mean age = 33.2 +/- 8.7 years) with a traumatic lesion (T1-L3) performed a graded exercise test on a wheelchair ergometer until exhaustion. The test started with an initial workload of 0 watts, with an increment of 6 watts per 2 minutes. Measurements included power output (W), heart rate (HR) and oxygen consumption (VO2) throughout the test. Linear relationships were observed between VO2 and W (VO2 = 0.79 + 0.02 W, r = 0.80, SEE = 0.22 l min-1) as well as between %VO2 max and % maximal heart rate (% VO2 max = 8.7 + 0.83 %HR, r = 0.83 SEE = 10.5%). Combination of the two equations for estimating VO2 peak led to a linear relationship between the estimated and measured VO2 peak. Nonetheless, the strength and accuracy of the prediction were low (r = 0.49, SEE = 0.29 l min-1). Participation in aerobic exercise, body mass and lean body mass, introduced as correction factors in the regression equation, significantly improved the strength and the accuracy of the prediction (r = 0.85, SEE = 0.29 l min-1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247602 TI - Evidence of autonomic dysreflexia during functional electrical stimulation in individuals with spinal cord injuries. AB - The purpose of the investigation was to examine the safety and efficacy of functional electrical stimulation (FES)-assisted hydraulic resistance training in improving cardiovascular fitness in persons with spinal cord injuries. The cardiopulmonary responses of 10 high spinal cord injured (SCI) and five able bodied (AB) subjects were assessed during three bouts of FES-assisted leg extension exercise. The protocol involved three 30-minute tests: (1) unloaded leg extension, (2) hydraulically-resisted leg extension (loaded), and (3) a reproduction of the unloaded and loaded protocols to measure cardiac output (Q). Pre-measurements were made of body mass, mean limb weight, maximal force output and maximal oxygen uptake (incremental arm ergometry). Oxygen uptake (VO2), minute ventilation (Ve), respiratory exchange ratio (RER), heart rate (HR), blood pressure (BP) were recorded before, during and after tests. There was a significant difference in VO2 max between SCI and AB subjects. Cardiac output significantly increased between the loaded and unloaded tests. The significant increases from rest to unloaded and loaded exercise pointed to the potential value of adding resistance to a leg extension training regime. Heart rate and BP of the participants with SCI consistently demonstrated a response suggestive of autonomic dysreflexia. Upon stimulation an immediate increase in (predominantly systolic) BP was observed, followed by a fall in HR. On cessation of stimulation HR exhibited a substantial rebound effect and BP returned to normal levels. This response was highly reproducible and suggests caution be exercised in the use of FES for people with SCI with lesion levels above the major splanchnic outflow (T6). PMID- 8247603 TI - Deep vein thrombosis in spinal cord injured patients. AB - In this study to determine the incidence of deep vein thrombosis (DVT) in spinal cord injury (SCI) patients, we evaluated 30 instances of bilateral ascending venography obtained in 31 patients. Every patient was on prophylactic low dose heparin anticoagulant therapy. The incidence of DVT was found to be 53.3%. Adverse effects due to venography were seen in 10% of patients. No major complications including postvenographic phlebitis and allergic reactions were observed. PMID- 8247604 TI - Errors and limitations of the multimodality checking methods of defective spinal intrathecal pump systems. Case report. AB - The loss of benefit from intrathecal baclofen, with increased spasticity and a discrepancy between the residual and the calculated volume content (underinfusion), made us suspect dysfunction of the intrathecal baclofen infusion in a paraplegic patient. Although all possible usual checking methods were used, no failure in the drug administration device (DAD) could be found. Despite some benefit from increasing the daily dose of baclofen, it has not been possible to control the spastic symptoms. Surgery was therefore decided upon, and a small leak at the end of the catheter tip was discovered. Surgeons and physicians should be aware that checking methods of DAD cannot exclude failure of the system. Their errors and limitations are discussed. PMID- 8247605 TI - Carcinoma of the scrotum associated with rubber urinals. Case report. AB - A paraplegic patient, who had worn a rubber urinal in close contact with the scrotum for over 40 years, developed carcinoma of the scrotum. It is likely that this is a previously unsuspected example of chemical carcinogenesis, and in view of this, we believe that the use of rubber urinals should be discontinued. PMID- 8247606 TI - Cytotoxicity of Porphyromonas gingivalis toward cultured human gingival fibroblasts. AB - Direct cytotoxicity of black-pigmented anaerobic rods was studied on the confluent monolayer of human gingival fibroblasts in vitro. Only strains of Porphyromonas gingivalis caused morphological alteration (cell-rounding) and notable depression of viability of fibroblasts. To determine the location of the cytotoxicity, bacterial surface components, i.e., outer membrane, lipopolysaccharide, fimbriae and outer membrane vesicles were prepared from P. gingivalis and their cytotoxicity was assessed. Among these preparations, only outer membrane vesicles are supposed to have high affinity to human gingival fibroblasts, and the cytotoxicity of outer membrane vesicles was found to be much stronger than that of the other constituents. This cytotoxic factor seemed to consist largely of protein and to be associated with the enzyme activity of outer membrane vesicles. The effects of some protease inhibitors and L-cysteine on the cytotoxicity of outer membrane vesicles suggest that the mechanism of cell rounding is different from that of cell death. PMID- 8247607 TI - Genetic transformation of Porphyromonas gingivalis by electroporation. AB - Porphyromonas gingivalis was transformed by electroporation using the DNA of plasmid pE5-2, or its derivative, pYT7. Prior to transformation, pE5-2 was transferred from Escherichia coli to P. gingivalis strains by conjugation (mobilization with R751), and the plasmid DNA was purified from the P. gingivalis transconjugants. Transformation occurred when the recipient strain and the donor strain from which the plasmid DNA was purified were homologous. If they were heterologous, transformation did not take place or did so at a very low frequency. This suggested that a restriction-modification system is present in P. gingivalis strains. Plasmid pYT7 was derived by removing an 8.0 kb AvaI fragment from pE5-2 that was purified from P. gingivalis cells. It has several single cutting restriction sites such as EcoRI, AvaI and ClaI usable for gene cloning, though it was not stable enough in P. gingivalis cells, probably because the rep gene was derived from a relatively distant species, Bacteroides eggerthii. PMID- 8247608 TI - Microbial associations in periodontitis sites before and after treatment. AB - Duplicate samples from 110 periodontal sites of 6 mm or more pocket depth in 16 patients were analyzed for the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Capnocytophaga spp., Campylobacter rectus, Eikenella corrodens and Fusobacterium nucleatum. The sites were sampled before and after nonsurgical periodontal treatment. No statistically significant associations were found before treatment between any of the analyzed species. After treatment, statistically significant associations were found between E. corrodens and all the other species, F. nucleatum and P. intermedia; Capnocytophaga spp. and C. rectus; P. intermedia vs Capnocytophaga spp. and P. gingivalis; and C. rectus vs Capnocytophaga spp. and A. actinomycetemcomitans. Some of these associations could be explained either by patient-related factors or site-related characteristics such as the pocket depth. The proportion of P. gingivalis seemed to be unrelated to the proportion of P. intermedia in the samples. If one of the analyzed microbes was found in one of the sampled pockets in a patient, the probability of finding that microbe in all the sampled sites in the same patient before treatment was more than 50%. This probability was reduced after treatment for many species, especially P. gingivalis, which showed a probability of zero. The probability of detecting a bacterial species on at least one additional site if it was present on one in the same individual was nearly 100%, both before and after treatment, for all species studied. This study has shown several potential microbial associations in the subgingival plaque flora of deep periodontal pockets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247609 TI - Porphyromonas endodontalis: prevalence and distribution of restriction enzyme patterns in families. AB - In this study we determined the prevalence and distribution of Porphyromonas endodontalis in 26 families consisting of 107 subjects. P. endodontalis was present in 24% of the investigated subjects and was recovered most often from the dorsum of the tongue (50%). Isolation was also possible from the tonsils, the buccal mucosa, the saliva and the periodontal pocket. The usefulness of restriction endonuclease analysis as a typing method for this particular species was investigated by typing 19 isolates from unrelated individuals. All these isolates had unique restriction endonuclease patterns. The observed heterogeneity indicates that restriction endonuclease analysis is a sensitive measure of genetic dissimilarity between P. endodontalis isolates and is able to characterize individual isolates. Application of restriction endonuclease analysis to the obtained clinical isolates in this study shows the possibility of the presence of multiple clonal types within one subject. The DNA patterns of all P. endodontalis isolates from unrelated individuals were found to be distinct. In 3 families the DNA patterns of isolates from the mother and her child were indistinguishable. These data indicate the possibility of intrafamilial transmission of P. endodontalis. PMID- 8247610 TI - Campylobacter rectus in human periodontitis. AB - Campylobacter rectus (formerly Wolinella recta) in periodontitis lesions was studied relative to age and sex distribution, relationship to disease-active periodontitis, response to periodontal debridement and in vitro antimicrobial susceptibility. Subgingival C. rectus was collected with paper points, transported in VMGA III and plated onto nonselective enriched brucella blood agar and Hammond's selective medium for C. rectus, both incubated anaerobically. C. rectus was recovered from 80% of 1654 periodontitis patients. Although the organism showed similar age and sex occurrence, its proportional recovery in culture-positive adults was inversely related to increasing age (r = 0.999, P < 0.001). The organism was positively associated (summary odds ratio = 2.95) with disease activity in a 24-month longitudinal study of 93 adult periodontitis patients on maintenance therapy. C. rectus decreased from 8.2% to 0.7% following local periodontal debridement of 20 culture-positive adult periodontitis patients. The organism exhibited high in vitro susceptibility to therapeutic levels of tetracycline hydrochloride, metronidazole, penicillin G and ciprofloxacin. These findings further delineate the epidemiology and potential pathogenic role of C. rectus in human periodontitis. PMID- 8247611 TI - Involvement of treponemal surface-located protein and carbohydrate moieties in the attachment of Treponema denticola ATCC 33520 to cultured rat palatal epithelial cells. AB - We studied the nature of attachment of Treponema denticola ATCC 33520 to a microscopically distinct population of rounded rat palatal epithelial cells. The motility of the freshly harvested spirochetes appeared not be a prerequisite for attachment. Treatment of T. denticola ATCC 3350 with proteinase-K, heat, glutaraldehyde, formaldehyde and periodate oxidation decreased the attachment to the rounded rat palatal epithelial cells, indicating the involvement of protein and carbohydrate moieties. Trypsin treatment had no effect on the attachment. The attachment of T. denticola ATCC 33520 was decreased after treatment with native non-immune rabbit serum, native polyclonal rabbit serum, D-mannose, N-acetyl-D galactosamine and sialic acid. The results indicate that the attachment of T. denticola ATCC 33520 to rounded rat palatal epithelial cells is mediated by trypsin-resistant adhesin(s) of protein and carbohydrate nature, with affinity for D-mannose, N-acetyl-D-galactosamine and sialic acid. PMID- 8247612 TI - Viscosity-dependent locomotion of oral spirochetes. AB - Spirochetes are thought to remain motile in environments (such as intercellular spaces) that immobilize extracellularly flagellated eubacteria. This attribute suggests that the viscosity of the milieu is of importance to locomotion. We sought to determine the interdependence of oral spirochete locomotion with media viscosity. Video time-lapse microscopy using darkfield optics was used. The motility of the spirochetes in media of different viscosities (various concentrations of Noble agar) was measured. Treponema denticola exhibited the fastest speed (18.7 +/- 4.4 microns/min) at a viscosity of 30 mPa.s. The highest speeds for Treponema vincentii and Treponema socranskii were 41.9 +/- 14.9 and 33.4 +/- 13.2 microns/min, respectively, at 88 mPa.s. These data show that optimal migration of spirochetes is viscosity-dependent. The results support the hypothesis that such viscosity-dependent locomotion could be a virulence factor that enables oral spirochetes to initiate and sustain periodontal disease. PMID- 8247613 TI - Binding of hemin and congo red by oral hemolytic spirochetes. AB - Colony-forming units or cells in suspension of oral anaerobic spirochetes (Treponema denticola, Treponema vincentii and Treponema socranskii) bind hemin and Congo red. Hemin or Congo red binds to a hydrophobic polypeptide receptor that is located in the outer membrane of the bacterial cells and it has a relative molecular mass of 47 kDa. These oral spirochetes also lyse sheep erythrocytes to produce beta-hemolytic zones around colony-forming units. The oral spirochetes may acquire iron for growth when they lyse erythrocytes and bind heme from which they may sequester and transport iron into the cells. PMID- 8247614 TI - Observation of an unusually large spirochete in endodontic infection. AB - Spirochetes in the root canal have been investigated only to a minor extent. Oral spirochetes have been detected particularly in the periodontal pocket, where small, medium-sized and large morphotypes have been reported. In the present darkfield and scanning electron microscopic study we describe a spirochete isolated from the root canal that is 7 times longer (140 microns long) and 5 times thicker (2 microns thick) than usual for treponemes. PMID- 8247615 TI - The role of reactive oxygen intermediates in the pathogenesis of chronic apical periodontitis. AB - The level of malondialdehyde, a stable end product of lipid peroxidation induced by reactive oxygen intermediates and the activity of two potent antioxidant enzymes, superoxide dismutase and glutathione peroxidase, was investigated in tissue homogenates of 22 surgical periapical granuloma specimens. Malondialdehyde levels were significantly higher and glutathione peroxidase activity was significantly lower in periapical granuloma samples than in healthy gingival tissue homogenates, which were used as controls. The activity of superoxide dismutase was similar in periapical granuloma and in control samples. Our results indicate an altered balance between the production and the elimination of toxic oxygen metabolites in chronic apical periodontitis. We hypothesize that reactive oxygen intermediates, which are being produced by activated phagocytic cells abundantly present in periapical granulomas, can contribute to periapical tissue injury and bone loss in this disease. PMID- 8247616 TI - Surgical management of equinovarus deformity in the adult with head injury. AB - Equinovarus is the most common residual deformity of the lower extremity in patients who have sustained intracranial injury. Appreciation of the pathophysiology of the deformity aids in surgical management of selected cases. Gait analysis is a valuable tool for preoperative management. Three case reports illustrate the principles of surgical treatment. PMID- 8247617 TI - Complete soft-tissue clubfoot release with and without internal fixation. AB - Two groups of patients who underwent soft-tissue release of clubfoot are compared. In group I, internal fixation (two Kirschner wires) was used for 6 weeks to retain correction of clubfoot. In group II, no internal fixation was used. Patients from the two groups were operated on by one surgeon using the same procedure. Because there was no statistically significant difference in functional result between the two treatment groups (P = .08; Mann-Whitney Test), the authors recommend avoiding internal fixation (with all the associated problems) in cases of uncomplicated clubfoot, and encourage early removal of the cast and frequent manipulations to prevent stiffness of the joints and reduce the recurrence rate. PMID- 8247618 TI - Soft-tissue window to enhance visualization of entrapped osteocartilaginous fragments in the hip joint. AB - The thin-section computed tomographic (CT) scans of 11 patients with entrapped intra-articular fragments secondary to dislocation or fracture dislocation of the hip are studied. The effects of CT bone and soft-tissue contrast manipulation on the visibility of the intra-articular fragments are demonstrated in both patients and in vitro. When the mechanism of injury and clinical features raise the possibility of entrapped fragments, as in hip dislocation and fracture dislocation, special magnification and image-enhancement techniques are available for re-examining the acetabulum, especially when raw CT data are saved. Thus, index of suspicion and appropriate CT planning can ensure prompt diagnosis and treatment of entrapped intraarticular fragments in the hip joint. PMID- 8247619 TI - The anterior humeral circumflex vessels and the axillary nerve. An anatomic study. AB - The anatomic relationship between the anterior humeral circumflex vessels and the axillary nerve was studied in 24 fresh-frozen cadaveric four-quarter shoulder specimens. Each shoulder was exposed through a standard deltopectoral incision, and the axillary nerve, anterior humeral circumflex vessels, and bicipital groove were identified. The perpendicular distance from the medial border of the bicipital groove to a point where the vessels crossed anterior to the nerve measured 2.6 +/- 0.4 cm (range, 2.1-3.6 cm). The perpendicular distance from the medial aspect of the bicipital groove to the point where the axillary nerve crossed the inferior border of the subscapularis and thus entered the quadrangular space was 3.0 +/- 0.5 cm (range, 2.1-4.4 cm). The anterior humeral circumflex vessels were easily located in all specimens, and the axillary nerve could be identified by its relationship to the vessels as described above, thus providing the operating surgeon with a consistent and reliable method of recognizing this nerve during anterior shoulder procedures. In addition, the axillary nerve was palpable in all specimens by sweeping the index finger in a medial-to-lateral direction along the inferior border of the subscapularis. PMID- 8247620 TI - Acute transphyseal hematogenous osteomyelitis caused by Haemophilus influenzae with 5-year follow-up. AB - Acute hematogenous osteomyelitis is a well-recognized entity in infants and children. Over 90% of cases are caused by coagulase-positive Staphylococcus aureus, with other organisms only rarely implicated. Transepiphyseal osteomyelitis, because of the presence of infection on either side of the physis, usually results in growth arrest. The authors report the case of a 4-month-old child who had a transphyseal acute hematogenous osteomyelitis caused by Haemophilus influenzae and document complete resolution of the infection at 5 year follow-up with no clinical or radiographic evidence of growth arrest. PMID- 8247621 TI - Isolated fracture-dislocation of the fourth carpometacarpal joint. A report of two cases. AB - Dislocations and fracture-dislocations of isolated and multiple carpometacarpal (CMC) joints are rare injuries that usually result from the impact of a clenched hand on an immobile object. Various authors have presented combinations of dislocations of any or all of the CMC joints. Isolated fracture-dislocation of the fourth CMC joint, however, is either exceedingly rare or highly underreported. Two cases of isolated fracture-dislocation of the fourth CMC joint are presented here: in both, the appearance of the hand suggested a fracture dislocation of the CMC joint, and the isolated lesion was confirmed at the time of surgery. Both patients were treated with open reduction and internal fixation and had good results at 2 years. PMID- 8247622 TI - A 21-year-old man with pain and swelling of the right hand. 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 this page. The final clinical and roentgenographic differential diagnoses are presented on the following pages. PMID- 8247623 TI - Inserting distal screws into interlocking IM nails--revisited. Methods to make it easier. AB - Inserting distal screws freehand is a difficult but necessary part of many intramedullary roddings. In the June 1992 issue of Orthopaedic Review, Edward Yang described an excellent protocol for performing this part of the procedure. Here, we describe more tips to make it easier and to prevent or correct problems. PMID- 8247624 TI - Imaging of the knee. PMID- 8247626 TI - On setting goals. PMID- 8247625 TI - Reduction of total hip dislocation. PMID- 8247627 TI - Pilon fractures of the distal tibia. AB - Pilon fractures are relatively rare fractures of the distal metaphysis of the tibia resulting from axial and/or rotational forces. They involve varying degrees of metaphyseal disruption, articular damage, and malleolar displacement. When severe, these injuries represent a tremendous challenge to the orthopaedic trauma surgeon. Traditional treatment of high-grade pilon fractures by closed means has led to unsatisfactory results. Open reduction and internal fixation in accordance with AO/ASIF principles has greatly improved the outcome in these often disabling fractures. Surgical technique involves anatomic restoration of fibular length, reconstruction of the plafond, bond grafting of the metaphyseal defect, and buttress plating of the medial tibia. Thorough preoperative planning and meticulous surgical technique produce predictably good results in the majority of cases. PMID- 8247628 TI - [Live attenuated viruses: a new strategy for vaccination against AIDS]. PMID- 8247629 TI - [Mechanism of allograft rejection]. PMID- 8247630 TI - [Enteroviruses and dilated cardiomyopathies]. PMID- 8247631 TI - [Detection and analysis of cryoglobulins: comparative study of a population of patients and one of healthy controls]. AB - During an 11-month period, 270 cryoglobulins were identified in 735 sera from patients versus 34 in 82 sera from apparently healthy blood donors. Western-blot analysis of the cryoglobulins was performed and cryoglobulin protein content was determined using a colorimetric assay. Comparison of findings in the two groups suggested that a substantial proportion of cryoglobulins identified in the sera of patients are quantitatively and qualitatively indistinguishable from cryoglobulins identified in healthy controls. PMID- 8247632 TI - [In vitro activity of glycopeptides on 114 coagulase-negative staphylococci: problems with the teicoplanin diffusion test, "alert" parameters indicating need for MIC determination]. AB - The activity of vancomycin (Va) and teicoplanin (Tc) against 114 coagulase negative staphylococci (CNS) strains was evaluated by disk-diffusion and agar dilution minimal inhibitory concentration (MIC) determination. Results were analysed by species and methicillin-resistance (MR) status. MICs for Va were in the susceptible range (< 4 mg/l) for all strains, with inhibition zone diameters of 17 mm or more in every case. MICs for teicoplanin were 8 or 16 mg/l for 19 CNS strains (including 18 with MR) of which 14 yielded an inhibition zone diameter of 17 mm or more (false susceptibility). Parameters which should lead to MIC determination for Tc regardless of disk diffusion findings include the species (S. haemolyticus), methicillin resistance, Tc zone diameter, and an at least 3 mm difference between the Va and Tc zone diameters. PMID- 8247633 TI - Electron microscopic description of accretions occurring in endotracheal tubes used in adults. AB - The purpose of the study was to describe the architecture of accretions occurring in endotracheal tubes used in adults by using a conservative procedure and transmission electron microscopy. The study included 12 tubes for which microbiological data of the tracheobronchial secretions were available. Observations were performed on inducted areas of the lumen. All tubes were covered with a several micrometers-to several millimeters-thick layer of mucus. The layers displayed stratified structures and showed granulations, neutrophils or cellular elements. When bacteria were seen, they made no contact with the polymer. This data suggest that adherence properties of bacteria towards the polymer were not involved at these stages of colonization and that a bacterial biofilms is a rare opportunity. PMID- 8247634 TI - Study of Rambach and NBGL agar on 4037 stools of human origin and 584 veterinary samples submitted for isolation of Salmonellae. AB - Rambach agar and novobiocin-brilliant green-glycerol-lactose (NBGL) agar are two recently described media designed for Salmonella isolation. Rambach and NBGL agars were compared to usual media in human and veterinary routines, by evaluation of sensitivities and predictive values of suspect colonies. In the human routine, 4037 stools, sensitivities were 79% for Hektoen agar, 51% for Rambach agar and 96% for NBGL agar at the direct plating for a total of 94 strains; and 89%, 87% and 94% respectively at the plating of Mueller-Kauffmann enrichment broth for a total of 143 strains. Predictive values of suspect colonies were respectively 58%, 93%, 91% at the direct plating; and 25%, 74%, 80% at the plating of Mueller-Kauffmann enrichment broth. In the veterinary routine, 584 samples, sensitivity was 65% on the association of Salmonella-Shigella and Hektoen agars, 70% on Rambach agar and 85% on NBGL agar, for a total of 40 strains. Predictive values of suspect colonies were 7.4%, 82.3% and 83% respectively. These results showed that Rambach and NBGL agars have distinct properties, and are both highly predictive. The highest sensitivities were achieved by the NBGL agar. PMID- 8247635 TI - [Cyclins A and B: redundancy and specificity]. AB - Mitotic cyclins constitute a regulatory subunit of the histone H1 kinase complex. On the basis of primary structure differences, they are divided into two classes, A and B. Both classes are necessary for mitosis to occur. Cyclins A and B differ in the timing of their cellular expression and in their affinity with the various members of the cdk (cyclin-dependent kinases) family. They also have specific functions: cyclin A plays a role in DNA replication, whereas cyclin B are involved in the inhibition of the fusion of early endosomes and in the activation of cdc25 phosphatase. Cyclins A and B can contribute to the development of neoplastic disorders, either directly (inappropriate expression of the cyclin A gene caused by the hepatitis B virus in hepatocellular carcinoma, interactions between cyclin A and factors involved in the regulation of cell division), or indirectly by causing phosphorylation of oncogene products by a cdk. PMID- 8247636 TI - [Clinical, diagnostic and therapeutic aspects of Moraxella catarrhalis infections]. AB - Moraxella catarrhalis is a Gram-negative diplococcus of the Neisseriaceae family now recognized as a cause of acute otitis media in pediatric patients and of lower respiratory tract infections in debilitated adults. The finding that 80% to 90% of strains produce beta-lactamases together with reports of cases of bacteremia and arthritis due to M. catarrhalis suggest an increasing pathogenic role for this organism. Antibiotic susceptibility testing suggests that the greatest bactericidal effect is provided by amoxicillin-clavulanic acid. PMID- 8247637 TI - [Evaluation of sensitivity of an assay: application to a third-generation thyrotropin (TSH) assay]. AB - The concentration-specific smallest detectable difference (D) between two concentrations (i.e., the sensitivity) was determined from the results of within run and run-to-run reproducibility studies of a third-generation thyrotropin assay. Only low concentrations were considered (range 0-0.3 m U/l). Assays were performed in singlet or in duplicate. Findings confirmed the value of the assay: the analytic detection limit approximated 0.005 mU/l and the functional detection limit was around 0.01 mU/l. Expressed as a percentage of the concentration, D% was under 20% for levels of less than 0.02 mU/l in within-run studies or of less than 0.03 mU/l in run-to-run studies. With concentrations in excess of 0.05 mU/l, D% was under 10% in within-run studies and slightly above 10% in run-to-run studies. These new detection thresholds for this assay should be taken into account when interpreting clinical results from a single run (TRH test) or from several runs (hormone replacement therapy or suppression test). PMID- 8247638 TI - [Programmed death of cells (apoptosis)]. AB - Apoptosis is a mode of cell death defined by characteristic morphological, biochemical and molecular changes. It was first described as a "shrinkage necrosis", and then this term was replaced by apoptosis to emphasize its role opposite mitosis in tissue kinetics. During apoptosis the cell decrease in size, loose contact with neighboring cells, and loose specialized surface elements such as microvilli and cell-cell junctions. A shift of fluid out of the cells causes cytoplasm condensation, which is followed by convolution of the nuclear and cellular outlines. In later stages of apoptosis the entire cell becomes fragmented, forming a number of plasma membrane-bounded apoptotic bodies which contain nuclear and or cytoplasmic elements. The ultrastructural appearance of necrosis is quite different, the main features being mitochondrial swelling, plasma membrane breakdown and cellular disintegration. Apoptosis occurs in many physiological and pathological processes. It plays an important role during embryonal development as programmed cell death and accompanies a variety of normal involutional processes in which it serves as a mechanism to remove "unwanted" cells. Apoptosis is associated with prostate atrophy after castration or atrophy of the adrenal cortex and thymus after administration of glucocorticoids. Apoptosis is involved in elimination of CD4 T lymphocytes in the course of HIV infection. The interest in apoptosis in oncology stems from the fact that it occurs in tumors, spontaneously as well as triggered by different antitumor drugs, radiation or after withdraw of growth factors. Spontaneous apoptosis may play a role in evolution of tumor malignancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247639 TI - Heterotopic induction of osteogenesis in the course of neural injury. AB - Encountered in orthopedics cases of accelerated bone union and abundant formation of the callus in patients after craniocerebral injuries as well as cases of extraskeletal ossification in neural diseases give rise to a question whether neural damage affects in any way the course of osteogenesis. The present study was carried out in an attempt to answer this question in an animal model. The study included 120 inbred WAG rats in which heterotopic induction of osteogenesis was performed by intramuscular placement of decalcified and lyophilized implants of rat cortical bones. By producing various neural damages their effect on the course of osteogenesis was evaluated. The rats were divided into four groups of 30 subjects each. In group I cerebral cortical damage was induced by intracortical injection of kainic acid solution, in group II paraplegia was produced by transverse dissection of the spinal cord on the level of Th 10-11, in group III denervation of the hind limb was performed by dissection of all supplying peripheral nerves, group IV consisted of controls. An attempt was made to evaluate changes in the course of osteogenesis while observing advancing with the passage of time changes in histological patterns of the preparations obtained from the site of bone implantation at 3, 5, 10, 20 and 40 days after the operation. While analyzing the histological pattern I paid attention to features characteristic for the process of osteogenesis such as: formation of granulation around bone grafts, penetration of mesenchymal cells into transplants, differentiation of mesenchymal cells into osteogenic cells, formation of bone tissue and development of bone marrow. The rate of development of these features in the consecutive preparations reflected the dynamics of induced osteogenesis. Analysis of the results showed that experimentally induced neural damage affects the course of osteogenesis. In case of cerebral cortical injury and peripheral neurotomy the formation of granulation around bone grafts was very abundant, whereas penetration of mesenchymal cells into the implants and differentiation of mesenchymal cells into chondroblasts occurred more rapidly than in other animal groups. In contrast, spinal cord injury resulted in a markedly decreased dynamics of osteogenesis which was manifested by weaker cell reaction around the implants and delayed differentiation of mesenchymal cells into chondroblasts. The effect was seen on the 20th day. In the final stage of the study--at 40 days--the effect of neural damage on the course of osteogenesis was reduced and the histological pattern was similar in all animal groups.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8247640 TI - [Humanitarian and academic aspects of animal experimentation]. AB - The author, while discussing the causes of increased demand for use of laboratory animals in biomedical research in the recent several decades, describes the origin of various social movements for the defense of the rights of animals in many countries. The paper lists the methods and ways of defending the animals' rights and numerous examples of appropriate resolutions and instructions of world institutions (UN, UNESCO, WHO), international and national bodies to regulate the animals' rights, protection and use for experimental and production purposes. The need for supervision of animal experiments, limitation of their use (only in indispensable and justified cases) and replacement with alternative methods for instance with cell, bacterial and fungal cultures, studies on species of a lower order, etc. is indicated. The author suggests that the number of laboratory animals in biomedical studies should be decreased emphasizing the need for appropriate organizational and administrative changes and use of animals with high sanitary and hygienic standards. The final part of the paper is devoted to nonspecific (environmental) anatomic and pathological changes found in the organs of laboratory animals. Based upon the proposed scoring system the author advocates a detailed description of these changes and comparison of the total pattern of changes with those in a matched control group. PMID- 8247641 TI - [Primary hyperparathyroidism in children]. PMID- 8247642 TI - [Ethical considerations on live transplantation from living related donors. Le conseil d'ethique de l'hopital Debrousse]. AB - This review examines the ethical problems of liver transplantation from living parental donors regarding the main ethical principles. It appears that this method of transplantation is a new possibility offered to patients that cannot be subtracted from the parents' choice without interfering with their liberty. Moreover, there is a social decisiveness to ensure the offering of this option which is limited by the conditions of realisation, timing and personal situations of the parents. The considerations of risks, benefits, advantages and prejudices between donor and recipient show a favourable balance for this type of transplantation provided that adequate medical indications are respected. However, the considerations of psychological cost and burden deserve special attention and are detailed in this paper especially regarding the motivations and the long-term effects on the donor. Futility and ethical generalisation examinations favour such a procedure. Moral justice will generally be respected. The consideration of autonomy required special attention owing to the situation pressure upon the donor's consent. Consent deserves adapted solicitation according to variable situations: whether the donor explicitly specifically wanted to donate part of his liver or only required information. Excessive solicitation of parents' participation to liver donation represents qualified "forced choice"; in some situations where the parents cannot effectively refuse their participation, paternalistic action is ethically acceptable. A general procedure is proposed to ensure the psychological, legal and ethical protection of the donor and recipient. PMID- 8247643 TI - [Selective bronchial occlusion in the treatment of unilateral interstitial emphysema in respiratory distress in premature infants]. AB - A case of right unilateral pulmonary interstitial emphysema in a compromised premature infant was managed by right selective bronchial occlusion with a favorable outcome. Occlusion with a Rashkind catheter occurred on day 8 and was well-tolerated. The balloon was deflated 10 hours later and no recurrence was observed. Right occlusion is easier than left selective intubation and seems to be the best method for treatment of unilateral interstitial emphysema in infants in extremely poor conditions. PMID- 8247644 TI - [Congenital deficiency of fibrin stabilizing factor [factor XIII)]. AB - The authors report a new case of congenital deficiency of fibrin stabilizing factor. The defect is transmitted as an autosomal recessive trait. The earliest symptom is umbilical bleeding when the cord separates. The diagnosis is confirmed by the rapid dispersion of the clot in urea 5 M or monochloroacetic acid. PMID- 8247645 TI - [3 pediatric cases of leptospirosis]. AB - Three children presented with an association of pains, infectious syndrome, acute renal failure, hepatitis and meningitis, that lead to the diagnosis of leptospirosis. The clinical spectrum of this rare disease are recalled. PMID- 8247646 TI - [Epstein-Barr virus infection and syndrome of inappropriate macrophage activation]. AB - An 11-year-old girl presented with a typical serologically proven infectious mononucleosis with persistent fever, jaundice and hepatosplenomegaly in spite of steroid therapy. Laboratory tests showed pancytopenia, fibrinopenia and hypertriglyceridemia. The liver biopsy revealed an infiltration with hyperbasophilic cells. One month later, a slight improvement was noted and fever disappeared after 4 days on acyclovir therapy. The authors recall the spectrum of the macrophagic activation syndrome. PMID- 8247647 TI - [Primary hyperparathyroidism in 2 adolescents]. AB - The authors report on two cases of primary hyperparathyroidism (PHP) in adolescents. In a 15-year-old boy, the clinical picture included skeletal, dental, renal and psychological manifestations. In the second case, the first symptom was urolithiasis at age 12; following a period without any important clinical event, the diagnosis was established when the patient was 18. In both cases, increased calcium and low phosphorus plasma levels associated to moderately elevated plasma parathyroid hormone levels were noted. CT scan and ultrasounds were normal but MRI showed an enlarged gland in the second case. The surgical exploration of the neck revealed an adenoma in the first case and a hyperplastic gland in the second one. Surgery was followed by an immediate improvement of both clinical and laboratory findings. These observations bring us to examine the spectrum of PHP in childhood, the difficulties in biological diagnosis and localizing techniques, and the recent results of surgical treatment. PMID- 8247648 TI - [Parathyroid adenoma in a child]. AB - We report the case of a 13 year-old white teenager who was referred for chronic bone pain. Physical examination showed a swelling of the anterior part of the neck and biological findings clearly evoked a primary hyperparathyroidism. The surgical treatment was successful and confirmed the diagnosis of parathyroid adenoma. Primary hyperparathyroidism being unusual in childhood, the eventuality of a multiple endocrine neoplasia is discussed. PMID- 8247649 TI - [Influence of sodium alginate on the intestinal transit in low birth weight newborn infants]. AB - Sodium alginate (Gaviscon) is used in the management of gastro-oesophageal reflux in infants. No digestive disadvantages have as yet been reported with the use of the Gaviscon formula available in France, which contains neither aluminium hydroxide nor thickener. Twenty-two healthy neonates were prospectively studied before and after Gaviscon treatment in order to characterize their whole gut transit time with the use of a carmine index. The head of the marker appeared within the same time in both experiments but the appearance of the tail was earlier in the treated infants (P < 0.05), without any subsequent clinical consequences. The slight increase shown in the rate of the clearance of the marker from the gut, is likely to be related to a less proximal to distal dispersion of the marker, subsequently to physical changes occurring in the viscous alginate. Frequency and consistency of the stools were unmodified by treatment and accordingly Gaviscon can be regarded as having no deleterious effect on transit time in neonates. PMID- 8247650 TI - [Pediatric urology from 1973 to 1993: diagnostic and therapeutic development]. AB - The author comments on the main aspects of the evolution of methods of diagnosis and treatment in pediatric urology during the past 20 years. In the field of diagnosis, ultrasounds have been a major break-through allowing early detection of uropathies and an easy supervision of their evolution. In the therapeutic field, improvements in surgical techniques and anesthesia have been numerous leading to both better results, and considerably reduced duration of hospitalization. PMID- 8247651 TI - [Psychiatric interventions in children under five years of age: prevention or care?]. AB - We analysed the consultations of children under the age of 5 attending our pedopsychiatric outpatients department over a period of 1 year. Although paediatricians and pedopsychiatrists agree on the necessity to start therapy as early as possible, we found that many children were seen to late, at a time when they had difficulties, and that the principle of early intervention in pedopsychiatric needs to be better applied during infancy in an effort to prevent psychological disturbances. PMID- 8247652 TI - [Chronic granulomatosis]. PMID- 8247653 TI - [Tuberculosis of the central nervous system: value of early polychemotherapy. Apropos of 2 cases]. PMID- 8247654 TI - Gender differences in locus of control scores for students with learning disabilities. AB - The present investigation examined gender differences on locus of control scores for students with learning disabilities. Scores from two measures of locus of control, the adult version of the Nowicki-Strickland Internal-External Scale and the Intellectual Achievement Responsibility Questionnaire, were compared for 77 boys (M age = 16.3 yr., M IQ = 89.5) and 27 girls (M age = 16.4 yr. and M IQ = 91.2). As groups were of unequal sizes, differences between scores were assessed by an analysis of unweighted means, creating a single harmonic mean and homogeneous subsets for all tests. Significant differences were found by gender on the Nowicki-Strickland inventory. On all assessments girls' means were more external than boys'. These results are discussed in light of recent findings concerning gender and positive adult outcomes for learners with learning disabilities and in relation to gender stereotyping and discrimination. PMID- 8247655 TI - Final comments on Hudesman, Page, and Rautiainen's (1992) subliminal psychodynamic activation experiment. AB - Hudesman, et al.'s (1992) contention that their finding shows that subliminal psychodynamic activation (SPA) improved academic performance is questioned. That experiment lacked controls outlined by Fudin in 1986 which are needed to support the assumption that a positive SPA outcome is effected because the meaning of an entire experimental message is encoded. In 1993 Hudesman and Page argued that Gustafson and Kallmen's 1991 results, obtained with such controls, indicated that the controls do not have to be used in subsequent SPA experiments. The 1990 results of Greenberg and of Kothera, Fudin, and Nicastro, however, do not support those of Gustafson and Kallmen. From a different perspective, it is argued that good experimental controls are needed in all SPA experiments because they increase internal validity. Given that Hudesman, et al.'s subjects scored in a limited range on the mathematics portion of the 1978 CUNY Skills Assessment Test, the implication that their result can be generalized to all subjects is questioned. PMID- 8247656 TI - Geophysical variables and behavior: LXXII. Barometric pressure, lunar cycle, and traffic accidents. AB - This study assessed relationships between traffic accidents and variables of the physical environment. Daily data on traffic accidents over a 4-year period were compared with daily records of barometric pressure and synodic lunar cycle. No significant variations in the number of accidents were found related to barometric height but a significant lunar periodicity was observed for one of the years considered. The number of accidents occurring during the full moon day was lowest; the highest occurred two days before the full moon. Accidents occurred more frequently during crescent moon than during waning moon, but no significant differences were noted when the lunar month was divided into four intervals of new moon, first quarter, full moon, and second quarter. PMID- 8247657 TI - Assessment of annihilation anxiety from projective tests. AB - This report details procedures to measure annihilation anxiety, a concept derived from Freud's 1926 formulation of traumatic anxiety. A 25-item pencil-and-paper inventory administered to patient and to nonpatient samples is described, along with a brief summary of earlier findings. The delineation of nine interrelated experiential components of annihilation anxiety provides the background for the construction of Rorschach and TAT measures of the concept. Findings comparing the pencil-and-paper inventory and the projective test measures are presented as well as examples of responses judged to reflect annihilation anxiety from Rorschach and TAT protocols. PMID- 8247658 TI - The sex ratio in samples of attempted and completed suicides over time. PMID- 8247659 TI - Stereophotometric testing for Pulfrich's phenomenon in professional baseball players. AB - Major League players were significantly more accurate in performing stereophotometry than were Minor League players. The stereophotometric data based upon the induction and extinction thresholds of the Pulfrich phenomenon were significantly correlated with the batting averages of Major League baseball players. The coefficient of determination, r2, implies that visual ability as measured by stereophotometry accounts for 47% or more of the variation in batting averages of the Major League players. This test may be a useful index in predicting batting ability. PMID- 8247660 TI - Prevalence of walking impairment in young, healthy adults. AB - 64 students' mean walking distance was greater than that of a presurgical group studied by Regensteiner and colleagues. The students were also faster. Between the distance and speed measures, r = .08. PMID- 8247661 TI - Perception and allocation of time by dyslexic children. AB - The perception and allocation of time by individuals is closely associated with their observed behaviors in life. A dyslexic child presents unique and interesting problems to educational researchers, teachers, and parents since the information-processing style differs from that of normal children. This study examined not the cognitive understanding of time or problems in measuring time (dyschronometria), but the perception and allocation of time by 26 normal and 21 dyslexic children between the ages of 10 years and 17 years old. These findings were measured by a specifically designed time allocation and perception questionnaire and a circles test. The analysis indicated that the dyslexic children tended to be far more outer- and inner-directed with regard to their time allocations and generally perceived time as a linking of past, present, and future events. PMID- 8247662 TI - Snoring and nightmares. AB - To test the conclusion of a recent study that snoring and disturbing dreams are strongly associated, 199 university undergraduates responded to a questionnaire that assessed their level of snoring and the frequency with which they experienced fantastic nightmares. Contrary to data provided by this recent study, we did not find a significant relationship between snoring and disturbing dreams. Some possible reasons for the discrepancy between these data sets were considered. PMID- 8247663 TI - Ascending-descending threshold difference and internal subjective judgment in CFF measurements of depressed patients before and after clinical improvement. AB - The purpose of this study was to measure, using the method of limits, the ascending-descending threshold difference, delta = fa-fd, in subjects with major depression. 28 patients were given two sessions, one before treatment and the other after recovery. Mean values of delta measured in the second session were significantly smaller than those scored before treatment. The two sets of values were positively correlated. The ascending-descending threshold difference was suggested previously as likely related to the internal subjective judgment of subjects. The present results are consistent with the hypothesis of a more conservative response criterion in depressed patients before than after clinical improvement. PMID- 8247664 TI - Evidence of a rapid matching process for simultaneous reading of several items from visual information storage. AB - When subjects searched for a digit which did not appear in an array of nine different, briefly displayed digits, they identified them more rapidly than Sperling reported earlier could be done. Subjects stated they could "see" the missing digit. We hypothesized that the "seen" digit would be the only remaining digit after subjects matched an internal set of 10 digits with the displayed set of nine stimuli. This process works early as sensory information storage and facilitates character identification. Here we presented another stimulus array immediately after the first one to examine the matching process by extinguishing the visualized digit. When the two arrays had different missing digits (Condition D), identification of the missing digit from the first array was worse than when they were the same (Condition-S). For the former condition, the remaining digit was extinguished after it was actually presented in the second array. The results support subjects' self-reports and clarify the matching process in the early information-processing stage. PMID- 8247665 TI - Body-shape perceptions of elderly women exercisers and nonexercisers. AB - Perceptions of current and ideal body shape for 49 elderly women exercisers (66 to 88 yr.) and 29 nonexercisers were measured using a nine-figure silhouette scale. Exercisers showed greater difference between current and ideal shape than nonexercisers, but no association was found for age. Further research needs to consider additional socioeconomic factors and be validated from a broader population. PMID- 8247666 TI - Light milk products: effects of product information on women participating in the Women's Ten Kilometer Run. AB - 418 women participating in the Women's Ten-kilometer Run completed a questionnaire of 51 questions. We hypothesized that a reliable health education brochure on light milk products would help to change consumption habits of milk products and other eating habits. 13% of women reported a positive change in their consumption of milk products. Seven variables explained 41% of the change in eating habits, and three variables (change of eating habits, knowledge about the prevention of osteoporosis, and reading the brochure) 37% of the change in consumption of milk products. PMID- 8247667 TI - Measuring physical self-efficacy in old age. PMID- 8247668 TI - Development differences in vowel duration in open and closed syllables. AB - Vowel duration in open and closed syllables was investigated in groups of ten normal 5-, 8-, and 10-year-old children, and adults. Productions of three syllables (bee, bead, beet), analyzed acoustically for vowel duration, indicated that for all age groups, mean vowel durations for "bee" and "bead" were significantly different from "beet" but were not different from each other. Developmental effects were noted in mean and variability of vowel duration. In addition, a context effect was observed, with adult-like values for both mean and variability obtained at an earlier age for "beet" than for "bee" and "bead." These results suggest that both phonemic and phonetic factors are important in accounting for developmental changes in mean and variability of vowel duration. PMID- 8247669 TI - The quality of life in urban areas and suicide. PMID- 8247670 TI - Cognitive functioning in male polydrug abusers with and without family history of alcoholism. AB - We investigated the family history of alcoholism as a potential contributor to cognitive dysfunction in polydrug abusers. A sample of 62 male polydrug users whose main drug of use was heroin included no one with which comorbid antisocial behavior varied by family history of alcoholism. They were compared on a battery of cognitive tests. Polydrug users with positive histories tended to perform worse on several cognitive measures than those with negative family histories, but these tendencies were not statistically significant. The current findings support the hypothesis that cognitive deficiencies observed in previous studies of persons with positive family histories of alcoholism may, in part, be attributable to a failure to exclude individuals with diagnoses of antisocial behavior. PMID- 8247671 TI - Examination dreams. PMID- 8247672 TI - Preschool embedded figures test performance of young children: age and gender differences. AB - An analysis of the literature was the basis for a set of predictions regarding the Preschool Embedded Figures Test performance of a small, cross-sectional sample of 37 3- to 5-year-old children. The test scores were modestly reliable. Predicted age-related differences in scores for boys and girls were observed, including an interaction of age with gender; however, other predictions regarding those scores were not supported. Based on a small sample, it was tentatively concluded that the evidence for continued use of the Preschool Embedded Figures Test as a measure of field independence for young children was weak. PMID- 8247673 TI - Effects of physical inactivity on all-cause mortality risk in Upper Bavaria. AB - The effect of physical activity on all-cause mortality was examined using a representative random sample of 1,536 persons (15 years and older) in three communities in Upper Bavaria. 27.0% of the respondents reported regular and 26.2% occasional physical exercise. During the 5-year follow-up 5.1% (n = 79) of the original sample died. Using a logistic regression model, the relation between physical activity and mortality was measured by the odds ratio, with subjects reporting regular physical activity as the reference group. Crude mortality risk was significantly higher among the physically inactive (men: 3.97; women: 4.36) but not among respondents practising occasionally (men: 1.67; women: 1.24). After adjustment for potential confounding variables (age, social class, physical and mental health), the mortality risk was elevated but not statistically significant for the physically inactive (men: 1.76; women: 1.51) and for the group practising occasionally (men: 1.50; women: 1.14). PMID- 8247674 TI - Recognizing suicides from photographs. PMID- 8247675 TI - Right-sided bias in traditional Kyokushinkai karate kata. AB - The 912 moves forming the 24 kata (forms) of Kyokushinkai karate were analysed for left-right bias. Significant right-sided bias was noted for counter-offensive techniques and over-all, but not for blocks. PMID- 8247676 TI - Effects of cognitive foveal load on a peripheral single-target detection task. AB - This experiment investigated the effects of foveal cognitive load on a primary peripheral single-target detection task. Four levels of foveal task with cognitive loads involving identification and summation of numerals were used. Number of correct targets detected seemed unaffected by the foveal load in the near periphery but a decrement occurred beyond 7.7 degrees. Response times for correct responses showed large dispersion compared with that for correct locations. At a low cognitive load, foveal task performance showed no deterioration for all eccentricities tested, but at a higher cognitive load performance declined gradually across eccentricities. Mild evidence of tunnel vision was obtained as indicated by the significant interaction of cognitive loads x eccentricities. Resources theory accounted well for the results. PMID- 8247677 TI - Suicide from bridges in Washington, DC. PMID- 8247678 TI - Young children's divergent movement ability. AB - To examine the relations of age, gender, movement experience, and gross motor development to young children's scores on divergent movement (i.e., ability to perform and modify fundamental movement patterns), 40 boys and girls ages 4, 6, and 8 years were tested. Ability to make divergent movements was evaluated by having subjects perform 3 different fundamental movement tasks. Movement experience was assessed by asking parents to complete a questionnaire. Ulrich's Test of Gross Motor Development was used to evaluate subjects' gross motor skill. Testing sessions were videotaped for analysis. Based on a forward stepwise regression analysis, experience and age accounted for 45% of the variance in children's gross motor skill. PMID- 8247679 TI - Heart-rate and blood-pressure responses to speech alone compared with cognitive challenges in the Stroop task. AB - Heart-rate and blood-pressure responses are assumed to reflect the "stressfulness" of cognitive tasks. Cardiovascular responses to speech are often assumed to be negligible. To test these assumptions, 34 middle-aged men (mean age 45.0 +/- 6.1) performed three versions of the Stroop color-conflict task, passive responding, push-button, and verbal. Although difficulty of passive responding was rated 11.8 (fairly light), push-button 16.1 (between hard and very hard), and verbal Stroop 14.5 (hard), all were significantly different. Analysis of variance showed during tasks heart-rate responses and systolic blood pressure did not differ. Recovery average heart-rate and over-all heart-rate patterns were not different for the difficult tasks but were significantly different from the easy task. Diastolic blood-pressure changes during tasks were more similar for verbal tasks despite the difference in difficulty. Stressor heart-rate and systolic blood-pressure responses did not reflect the difficulty of this stressful task. Verbalization of responses contributed significantly to cardiovascular reactivity. PMID- 8247680 TI - Visual figure-ground and perceptual stability. AB - A conceptual model that relates all forms of multistable figures is presented. It has the form of a tetrahedron whose faces and edges represent the variables of information contained in the ground, in the figure-ground relationship, in the extent of organization of the ground, and meaningfulness. PMID- 8247681 TI - Comparison among various methods of assessment of impulsiveness. AB - The current confused status of the research on impulsivity may be attributed to the lack of precise definitions, the reliance of most operationalizations on a single index, and inconsistency among different measures of the construct. Empirical measurements of impulsivity by self-reports, rating scales, or performance tasks suggest that the instruments employed measure aspects that have very little in common, a finding that throws serious doubts on the validity of the construct and implies a need for further research. To clarify this topic, we applied four different measures of impulsivity to 46 7th-grade (12 to 13 years old) schoolchildren. The children were rated by their teachers on an impulsivity behavior scale and were administered Kagan's Matching Familiar Figures Test, Version MFF-20, and two self-report forms, the Eysenck Impulsiveness Questionnaire, and the Barratt Impulsiveness Scale. Although the results confirmed the lack of convergence among these measures, high latencies on matching were associated with the cognitive aspect of the self-report scales. Treating impulsivity as a multidimensional construct is discussed. PMID- 8247682 TI - Androgyny predicts empathy for trainees in medicine. AB - Data from 65 medical students and residents support the hypothesis that scores on a measure of androgyny are predictive of those on an index of empathy but relatively modest predictive accuracy was observed for sympathetic responses. Further exploration is suggested. PMID- 8247683 TI - Group Embedded Figures Test: psychometric data documenting shifts from prior norms in field independence of accountants. AB - This study examined the performance of 516 practicing accountants on the Group Embedded Figures Test. The present accountants scored significantly more field independent than the published test norms and the 124 accountants in Pincus's 1985 sample. Significant differences were also noted between traditional (entry immediately following collage at age 22) and delayed-entry (late entry or second career) accountants. Gender differences noted in the norms were not observed for the current sample. PMID- 8247684 TI - Perceptual adaptation in the vestibulo-ocular system: EEG correlates of spatial and temporal rearrangement. AB - Spatial-temporal visuomotor rearrangement caused pursuit eye movements to counteract the vestibulo-ocular reflex (VOR). Vertical head nodding produced horizontal oscillations of a light spot delayed 1 or 150 msec. Adaptation resulted in apparent complementary motion of a stationary stimulus during nodding. 16 subjects adapted and tested at 150 msec. showed a 10% magnitude apparent motion. Following normal vision, while the electroencephalogram (EEG) was recorded, subjects were readapted at 150 msec. but tested at 1 msec. (to measure temporal generalization). Individual performance was correlated with EEG alpha. Adaptation correlated negatively with O(z) and Fz intensity, and positively with O(z) frequency. Temporal generalization correlated positively with O(z) intensity and negatively with O(z)-Fz phase angle. These results suggest that visuomotor adaptability is related to electrocortical activity. PMID- 8247685 TI - Language, personal-social abilities, and motor performance in preschool children and family characteristics of addicts in southern Italy. AB - To replicate a previous study on the same argument by Powers, Duran, and Reynolds (1993), 38 substance abusers' scores on family cohesion, expression, conflict, and the abilities of their children (language, personal-social abilities, fine and gross motor performance) were analyzed. The nonsignificant results confirm those found by Powers, et al. PMID- 8247686 TI - Type of stressor and depressive mood in patients with rheumatoid arthritis. AB - The relationship of two types of stressors to depressive mood was investigated in 54 patients with rheumatoid arthritis. Depressive mood was associated with disease-related stressors (pain, effects of disease on daily activities) but was unrelated to major life events. PMID- 8247687 TI - Spatial displacement among pedestrians: a cross-cultural replication. AB - An observational study of pedestrians in a shopping mall in a mid-western U.S. city was replicated in shopping malls in London. The avoidance movements of 2692 shoppers were recorded as they approached other shoppers. The relationships between these movements and the shoppers' gender, age, race, group size, and other characteristics were examined. Results of both studies indicated that men were likely to be displaced by women, smaller groups by larger ones, and nonhandicapped persons by handicapped ones. The results differed, however, with respect to race, with black persons tending to displace white individuals in the Midwest regardless of group size, while lone white pedestrians tended to displace lone black pedestrians in London. In London larger groups tended to displace smaller ones regardless of race. Hypotheses regarding spatial displacement were discussed. The need for more cross-situational and cross-cultural research in social psychology was clear. PMID- 8247688 TI - Magnitude and reliability of measurements of muscle strength across trials for individuals with mental retardation. AB - The purpose of this investigation was to examine the magnitude and reliability of strength measures on the Nicholas Manual Muscle Tester for individuals with mental retardation. 17 subjects were given six trials with the dominant and nondominant flexor and extensor muscle groups. No significant differences were found between dominant and nondominant muscle groups by side or by muscle group. Interclass correlations ranged from .96 to .98 for six and three trials, respectively. It was concluded that the test was appropriate for reliably assessing strength in this population and that these individuals can achieve a sufficient magnitude on three trials for a reliable measure of strength. PMID- 8247689 TI - Intelligence and motor skill acquisition by discrimination learning. AB - This experiment investigated the relation between intelligence and acquisition of motor skill using predictions from Zeaman and House's 1963 attention theory. 20 undergraduate students and 20 subjects of low IQ (WAIS-R Full Scale IQ range 50 to 73, M = 60) made linear positioning movements of long and short amplitude to the left and right of a central starting position. Four conditions (right-long, right-short, left-long, left-short) were created by specifying the corresponding target area on the positioning apparatus. One or both of the paired cues were varied, i.e., left, right, long, or short, while the dimensions of direction and amplitude remained unchanged across conditions. A shift from one condition to the next followed a criterion response of four consecutive movements to the target area. Results supported the hypothesis that subjects of low IQ would require more trials to criterion than subjects of normal IQ across all conditions of direction and extent. There was qualified support for the hypothesised interaction between intelligence and cue shifts. The practical implications and theoretical significance of these findings are discussed. PMID- 8247690 TI - Children's and adults' learning of a visuomanual coordination: role of ongoing visual feedback and of spatial errors as a function of age. AB - The role of ongoing visual feedback and of spatial errors in learning a target aimed task was investigated in 6-, 8-, and 11-yr.-old children, and young adults. In each of the four age groups, 16 subjects were randomly assigned to one of three conditions of learning (either with or without vision of the limb or with vision of final spatial error). All the subjects were tested without vision of the limb before and after the learning session. Before learning, directional bias was larger for 8-yr.-old children and target undershooting was greater for adults. After learning with vision of final spatial error, amplitude and direction improved for all age groups. Nevertheless, directional accuracy during the learning session increased more slowly for children than for adults. After learning with vision of the limb, direction improved for 6- and 8-yr.-old children only, demonstrating a specific role of ongoing visual feedback during learning at these ages. Further, an analysis of movement paths suggests a poorer capability for integrating proprioception by children than by adults. PMID- 8247691 TI - Development of the Health Student Self-efficacy Scale. AB - The Health Student Self-efficacy Scale assesses self-efficacy beliefs of students who are studying in courses allied with medicine. The psychometric properties (alpha = .81; several validities, r = .18, .44) indicated that the scale might be of use in monitoring performance of students in health-related courses. PMID- 8247692 TI - Effect of physical exercise on the performance of cognitive tasks. AB - This experiment examined the effect of physical exercise on measures of cognitive performance, Raven's Matrices, and an adaptation of the revised WAIS arithmetic subtest. We also tested the inverted-U hypothesis of an interactive relation between exercise-induced arousal and cognitive performance. 50 physically active men were assigned to five groups (n = 10) of equal physical fitness based on predicted maximum oxygen uptake. Three exercise groups undertook bench stepping at mean power outputs of either 47, 75, or 120 watts. One control group played Bingo and another control had no activity. There was no change in the Raven's Matrices scores pre- to posttest intervention, neither were there any between group differences either pre- or posttest. The arithmetic scores were significantly higher over-all (p < .05) on the posttest, but there were no reliable differences between groups either pre- or posttest. These results suggest that short duration (6 min.) aerobic exercise has no effect on cognitive performance. This finding supports the majority of previous studies that used step-up tasks to examine the relation between physical exercise and cognitive performance. PMID- 8247693 TI - The relationship between empathic response and scores on the California Psychological Inventory. PMID- 8247694 TI - WISC-R IQ estimates from the Luria-Nebraska Neuropsychological Battery. AB - Multiple regression equations predicting WISC-R IQs from Luria-Nebraska T scores and subjects' chronological age were developed and cross-validated on a total sample of 152 adolescent psychiatric patients. Data from a randomly drawn subsample of 100 subjects were entered into three stepwise multiple-regression analyses to derive equations predicting WISC-R IQs. These equations were applied to the remaining subsample (n = 52) as were the equations developed by McKay, et al. (1981) for estimating adult WAIS IQs. Mean differences, correlations, and classification hit rates between the two sets of predicted IQs and the actual WISC-R IQs were examined. Findings were similar to those of previous studies examining the predictive relationship between the Luria-Nebraska and adult intelligence. As expected, chronological age entered significantly into all three stepwise multiple-regression analyses. The newly derived equations had significantly higher correlations with the WISC-R for Verbal and Full Scale IQs and significantly better predictive accuracy for Full Scale IQ. Rates of IQ misclassification warrant caution in the use of these methods, but it is concluded that the equations derived here predict about as well as do short forms of the WISC-R. PMID- 8247695 TI - Asymmetries of manual motor response in relation to age, sex, handedness, and occupational activities. AB - Questions about left- and right-hand asymmetries require the analysis of preference and manual skill with respect to handedness. Hand preference and hand skill were examined in relation to sex, age, and occupational activities. 512 adults from manual, mixed, and intellectual activities and 253 children were tested for their manual preferences and performance. Analysis of variance followed by Scheffe post hoc tests, for right-handed adults, indicated significant effects for occupational activity and sex in dexterity and strength. Manual workers, under technological pressure, seemed more consistent in their manual preferences than their peers. For left-handed persons there were no significant effects. Dexterity and strength were also related to sex and age, suggesting different cultural opportunities for both sexes in relation to motor activities. PMID- 8247696 TI - Talent, special ability, and hemispheric processing. AB - The role talent training and special ability may play in hemispheric functioning is discussed. Further research on hemispheric processing from the perspective of cognitive strength as well as deficit is suggested. PMID- 8247697 TI - It's a long story: Public Speaking 301. PMID- 8247698 TI - Conducting a learning needs assessment. PMID- 8247699 TI - Nursing research. A qualitative perspective. Second edition. PMID- 8247700 TI - In another world: a phenomenological perspective and discovery of meaning in mothers' experience with death of a wished-for baby: doing phenomenology. PMID- 8247701 TI - Qualitative approaches in nursing research. Grounded theory: the method. PMID- 8247703 TI - Qualitative approaches in nursing research. Ethnography: the method. PMID- 8247702 TI - People with bipolar disorders quest for equanimity: doing grounded theory. PMID- 8247704 TI - Qualitative approaches in nursing research. Case study: the method. PMID- 8247705 TI - Qualitative approaches in nursing research. The centrality of caring: a case study. AB - This chapter has provided a rudimentary example of an exploratory/descriptive case study, utilizing phenomenological methods of data analysis. As discussed in the previous chapter, clearly delineated methods for case studies are not prevalent in the same way that other research methods are. The clear and fundamental design of this case study was purposeful as a demonstration of the utility of the method in day-to-day nursing. The aesthetic quality of the case study method is its simplicity of design. It is critical to note that the simplicity does not decrease the rigor or scientific merit of the process or outcome; in fact, an opposite argument could be made. The reporting style utilized was traditional so that the method could be clearly understood and compared to other research methods. PMID- 8247706 TI - Qualitative approaches in nursing research. Historical research: the method. AB - The use of the historical research process in nursing is a valuable approach to expanding nursing's understanding of itself, and for interpreting the field and its contributions to others. It provides a scholarly means of connecting the field to the whole of human experience. Its liberating and liberalizing quality assists the profession further to define its identity through understanding of its heritage and to provide direction for its future. As a research method, it links nurse scholars with their colleagues in the humanities. As a scholarly pursuit within the professional field, historical inquiry, properly executed, has become essential to the refinement of nursing's understanding of itself. PMID- 8247707 TI - Epistemology in nursing. PMID- 8247708 TI - Institutional review of qualitative research proposals: a task of no small consequence. PMID- 8247709 TI - Qualitative research proposals and reports. AB - In essence, a well-developed qualitative proposal provides: 1. A clear statement that specifies the phenomenon to be studied; 2. Documentation of a need for study, with specification of the significance of the study for nursing (or social science in general); 3. Acknowledgement of the researcher's a priori orientation(s) and perspective(s) with articulated questions about the attendant presuppositions; 4. Identification of the qualitative approach with a rationale for its selection; 5. Specification of the design with attention to: a. what data are sought, b. how and when those data will be solicited, c. how relationships with research participants will be initiated, maintained, and terminated, d. how data will be managed, e. how data will be analyzed and related to preexisting knowledge, f. how findings will be reported. PMID- 8247710 TI - Combining qualitative and quantitative approaches. PMID- 8247711 TI - Evaluating qualitative research. PMID- 8247712 TI - Language and nursing research. AB - The intent of this chapter can be summarized by borrowing Paterson's (1978) words: For responsible, effective existence the professional requires language [emphasis added] to relate authentically the purposes, beliefs, concerns, and events experienced continually in the nursing world. (p. 51). The noumenal exists in those phenomena listed by practicing nurses, but each seems to be a thing in itself--something waiting for description to bring it into our everyday awareness and to give it significance. It is as though we need to assert these events as nursing's, articulate our authentic experience with patients, claim what we believe is paramount to health (i.e., good nursing), and conceptualize what is uniquely the abstract and the concrete, the enduring and the relevant meanings of shared human experience between patient and nurse. We believe that qualitative research methods have much to offer as a research paradigm that is congruent with nursing's larger worldview, paradigm, or model. We close this chapter with Table 1-9, an illustration of the language of the qualitative research methods, and leave our readers to draw their own conclusions. PMID- 8247713 TI - Philosophical foundations of qualitative research. AB - Although new and still emerging for us, qualitative research approaches have been receiving considerable attention for some time in other disciplines. Along with philosophical debates, there are debates about whether there needs to be a debate. On a philosophical level, there is irreconcilable conflict between the quantitative and qualitative paradigms. It is important to recognize this conflict, avoiding illogical compromise. Yet, proponents of each paradigm need to applaud both the existence of the other and the hybrid paradigms that inevitably are born of conflict. An apt beginning would be broader definitions of what constitutes science and research in nursing, eliminating the sense-organ bias that is so contrary to our philosophy for practice. This alone would provide qualitative nurse researchers with the sanction they need to progress in their exploration of various approaches to creating a science and a body of knowledge in, for, and about nursing practice. In the chapters to follow, readers will be introduced to several qualitative research approaches. Each approach represents an interpretation of the qualitative paradigm in nursing research, grounded in the general perspective of phenomenological philosophy. This perspective focuses on phenomena as they appear and recognizes that reality is subjective and a matter of appearances for us in our social world. Subjectivity means that the world becomes real through our contact with it and acquires meaning through our interpretations of that contact. Truth, then, is a composite of realities, and access to truth is a problem of access to human subjectivity. This perspective guides the qualitative researcher in nursing to the subject matter of lived experiences, which are the original contacts with a world, and of the processes and content of interpretation--the meaning attributions that constitute realities and perspectives for a future of possibilities in the world. Other consequences of a phenomenological perspective in research include deliberate attention to the researcher's involvement in the study, engagement of multiple modes of awareness, and creative expression of findings. The product of efforts to establish a phenomenological baseline, a thorough and accurate description of nursing phenomena (a task that remains forever incomplete), will be clarified nursing concepts. If we encourage our qualitative nurse researchers, we can look forward to enhanced relevance in theoretical and empirical comments about nursing from studies guided by a mature nursing identity. PMID- 8247714 TI - Qualitative approaches in nursing research. Phenomenology: the method. AB - Anderson (1989) argued that the proper focus for phenomenological nurse researchers is working out the problems and dilemmas of doing phenomenology well, rather than pondering the possibilities of triangulated research. Among those problems are questions concerning how and when one "brackets," how the researcher's self is featured, and the issue of rigor in analysis of phenomenological data. In accord with this position, the overview of phenomenological method in this chapter reveals a need to continue development of a phenomenology that produces nursing knowledge to guide nursing practices. It has been proposed that the qualities and characteristics of the researcher researched relationship are particularly important to reconsider. The commonalities of the various phenomenologies that are engaging nurse researchers have been emphasized; their variations serve to raise methodological questions, redirecting us to yet another look at philosophical foundations for our science. The very subject matter of phenomenology is described variously. Some phenomenologies specify that the subject matter is such experiences as being in pain, doubting treatment, hoping to be well again, recovering from illness, or preparing for discharge. However, it may also be the mode of doubting or hoping, or it may be a focus on ways that it is possible to be in the world, such as what it is like to parent a child with a chronic illness, for example. The product of phenomenological inquiry, the description of experience, is inescapably interpretive, but the extent of the interpretation is another variability among the phenomenologies. However, to the extent that the description is an effective communication of insights into human experience, its relevance is immediate and direct. As a particular qualitative approach in nursing research, phenomenology has the distinction of turning the researcher on the self, that is, on his or her own reflective and intuitive grasp of experience. The researcher's direct experience constitutes the data, whether immediate or vicarious. It also has the distinction of deliberate modification by nurses for its use in nursing. This is a fortuitous development well worth our attention as the qualitative paradigm in nursing continues to gain momentum in revolutionizing our science and our practice. PMID- 8247715 TI - [Should Helicobacter pylori infections be treated?]. PMID- 8247716 TI - [Diagnosis of migraine in general practice]. AB - 195 patients with headache were examined by general practitioners and neurologists, and their diagnoses were compared. The general practitioners diagnosed migraine in 116 cases, the neurologists in 144 cases, and in 104 cases their diagnoses agreed. Thus, migraine can be difficult to diagnose, and this study indicates underdiagnosis of this common condition by general practitioners. This may result in overuse of analgesics and alternative therapies, and too little prescription of specific migraine medication. Greater familiarity with the diagnostic criteria of the International Headache Society should make it easier to arrive at a correct migraine diagnosis. PMID- 8247717 TI - [Antibiotic-induced diarrhea and pseudomembranous colitis]. AB - Although not all antibiotic-induced diarrhoea is pseudomembranous, and not all pseudomembranous colitis is antibiotic-induced, by far the greater proportion of intestinal lesions and diarrhoea occurring in conjunction with Clostridium difficile infection are attributed to previous antibiotic treatment. The article consists in a review of pathogenic, diagnostic, treatment and nosocomial aspects. PMID- 8247718 TI - [Hvidovre Hospital. Measures to control the spread of imported multiresistant Staphylococcus aureus]. AB - Methicillin-resistant Staphylococcus aureus (MRSA) are endemic in many hospitals abroad. Patients transferred from these hospitals may give rise to a spread of MRSA in Scandinavian hospitals, where MRSA only occur sporadically. A spread of MRSA may have great therapeutic and economic consequences for the local health care system and should therefore be met by strict hygienic measures. In Hvidovre Hospital patients transferred from or recently admitted to hospitals outside Scandinavia, are screened for MRSA. Patients carrying MRSA are isolated and discharged as soon as possible. Attempts are made to eliminate the carrier state by means of chlorhexidine. PMID- 8247719 TI - [Osteoporosis treatment--state of the art]. AB - In a number of Scandinavian studies it has been estimated that the incidence of femoral fractures in Scandinavia will double during the coming 20 years. Femoral fractures is also an increasingly frequent problem in developing countries. No treatment is available that will promote the regeneration of bone tissue. At best, remaining bone tissue can be strengthened. The article provides a survey of possible ways to prevent osteoporosis that are available now or may be introduced in the future. PMID- 8247720 TI - [Quality assurance in a medium-sized central laboratory]. AB - In the article are described the procedures and materials used at a medium-sized Danish central laboratory, vis-a-vis current trends in quality assurance. There is a tendency toward stricter control procedures. The development of bedside and office tests will entail a shift in the onus of quality assurance to individual physicians in the future. PMID- 8247721 TI - [Ethical and psychological aspects of screening tests for children]. AB - The increase in the incidence of screening procedures has been accompanied by increased awareness of the ethical and psychological problems involved, particularly in conjunction with the detection of diseases in children. Before a screening programme is launched, the ethical and psychological consequences for children and their families should be evaluated, as well as what supportive measures may be necessary to minimize the psychological effects of positive and false-positive test results. Moreover, it should be carefully determined which age is the most appropriate for screening with a view to the possibility of initiating prophylactic treatment and minimizing the adverse effects of stigmatization. PMID- 8247722 TI - [Physicians in former Yugoslavia build new bridges with Scandinavian support]. AB - For the first time since the war started in former Yugoslavia physicians from the medical associations in Bosnia-Herzegovina, Croatia, Macedonia, Montenegro, Serbia and Slovenia met in Oslo, Norway, to establish a dialogue between medical professionals on issues related to humanitarian assistance, health and medical issues. The three days meeting was initiated by the Norwegian Medical Association's Committee on Human Rights and financed by the Norwegian Ministry of Foreign Affairs. The meeting was held in cooperation with the World Health Organization and the medical associations of all the Nordic countries. The meeting in Oslo started in tension but ended with a joint statement (page 303) from all eleven participating delegations to continue dialogue on issues related to humanitarian assistance to the people of former Yugoslavia and to find ways of moving forward in health and medical issues. PMID- 8247723 TI - Control of properties of oligonucleotides by chemical modifications. PMID- 8247724 TI - The synthesis and some properties of some 5-substituted pyrimidine 4'-thio-2' deoxynucleosides and oligodeoxynucleotides containing 4'-thiothymidine. AB - A series of 5-substituted pyrimidine 4'-thio-2'-deoxynucleosides has been synthesized and their antiviral properties determined. It seems likely that once an analogue is a kinase substrate, then that analogue is toxic for that organism. Thus 4'-thiothymidine is phosphorylated by both viral and host kinases and shows toxicity to viruses and the host. Unlike its oxygen-containing counterpart (5 vinyl-2'-deoxyuridine, which is extremely toxic in vitro and causes chromosome damage), 5-vinyl-4'-thio-2'-deoxyuridine is not toxic and shows a similar antiviral activity spectrum to 5-cyclopropyl- and 5-isopropyl-4'-thio-2' deoxyuridines which also show no toxicity. Although a good leaving group at the 5'-position of a 4'-thionucleoside appears to be easily displaced (possibly involving intramolecular episulphide formation), when incorporated into an oligodeoxynucleotide, 4'-thiothymidine appears not to cause any gross distortion of the helix as shown by CD or Tm measurements and the phosphodiester backbone is stable to hydrolysis. Thus it is not yet clear whether the toxicity of 4' thiothymidine is due to its presence in DNA or to the perturbation of the metabolic processes involved in its incorporation. PMID- 8247725 TI - Mutagenesis, structure and function studies of thymidylate synthase. AB - The mechanism and structure of thymidylate synthase (TS) are fairly well understood. The function of amino acid residues of the enzyme have been studied by "replacement set mutagenesis" where multiple mutations at a given residue are produced and studied in detail. Some surprising results are (a) that there are few truly essential amino acids in TS, (b) that water may play an important role as a general acid/base catalyst, and (c) that a single mutation of Asn 229 to Asp changes specificity from a dUMP methylase to a dCMP methylase. Other mutagenesis and structure-function studies of interest will be discussed. PMID- 8247726 TI - Ribonucleotide reductases: radical enzymes with suicidal tendencies. PMID- 8247727 TI - Re-engineering DNA: design, synthesis, and properties of modified nucleic acids. PMID- 8247728 TI - Cloning of VH and VL genes for antibodies specific for pyrimidine photo dimers. AB - We have prepared two types of cDNAs encoding mouse immunoglobulin heavy (VH) and light (VL) chain variable (V) domains against UV-induced DNA damage by the polymerase chain reaction (PCR). The amplified cDNA was cloned for sequencing and expression, and the nucleotide sequences of the V domains were determined. The deduced amino acid sequences of the complementarity-determining regions (CDRs) indicated that CDR3 of the VH domain and CDR2 of the VL domain might be concerned with the recognition of photolesions. PMID- 8247729 TI - Design of hammerhead ribozymes for the inhibition of gene expression ex vivo by exogenous application. PMID- 8247730 TI - Structural studies of catalytic RNAs by heteronuclear multi-dimensional NMR. PMID- 8247731 TI - Chemical probing studies of the hepatitis delta virus (HDV) genomic ribozyme. AB - We have investigated the higher-order structure of HDV genomic ribozyme using various base-and phosphate-specific chemical probes under native, semi-denaturing and denaturing conditions. The bases of the HDV ribozyme were probed by treatment with dimethylsulfate and carbodiimide. In general, our results are consistent with the proposed pseudoknot model of secondary structure. The involvement of phosphates in hydrogen bonds or ion co-ordination was monitored with ethylnitrosourea. PMID- 8247732 TI - Multiple site-specific cleavage of HIV RNA by transcribed ribozymes from shotgun type trimming plasmid. AB - We have constructed a multiple-ribozyme expression system (designated shotgun type expression system). This system made it possible to express multiple ribozymes without reducing of the activities of each ribozyme. By using this system, multiple ribozymes specific for five different target sites in HIV-1 RNA were transcribed; all of them functioned independently of the others, and each cleaved the RNA at the respective predetermined target site only. Transient HIV-1 infection assay revealed that this kind of construct can disarm AIDS virus up to 96%, at least in a cultured cell. This proves that our system is effective in vivo as well. PMID- 8247733 TI - Sequence specific interaction of DNA with water-soluble porphyrins. AB - Footprinting experiments and restriction enzyme inhibition work as well as affinity cleavage studies have been carried out to probe the sequence specific recognition of DNA by water-soluble free-base porphyrins and their metal complexes. Porphyrins lacking functional groups capable of forming hydrogen bonds exhibited high DNA sequence specificity. Induced circular dichroism (CD) spectroscopy was found useful in analyzing the complex DNA binding modes of these compounds. PMID- 8247734 TI - Chemistry of oxidative DNA strand scission. AB - In order to investigate the reactivity of uracilyl-5-yl radical in fixed DNA structure, photoreaction of 5-halouracil-containing oligonucleotides was investigated. It was found that photoirradiation of BrU-containing duplex DNA provided C1' oxidation product selectively at adenine residue of ABrU sequence. In contrast, photoirradiation of IU-containing DNA gave C1' and C2' oxidation products with almost no sequence preference. Sequence specific electron transfer in duplex DNA is proposed in the photoreaction of ABrU. PMID- 8247735 TI - Photochemical cleavage of DNA by phosphorus(V) porphyrin derivatives. AB - Photocleavages of double-stranded circular DNA and oligonucleotide were investigated by the use of phosphorus(V)tetraphenylporphyrin (P(V)TPP) derivatives. The P(V)TPP derivatives cleaved the DNA and the oligonucleotide under the visible light irradiation, in air and in argon, respectively. It is suggested that the photocleavage reactions include an electron transfer mechanism as well as a singlet oxygen (1O2) one. PMID- 8247736 TI - Purification and characterization of RNases from fission yeast under nitrogen starvation. AB - Two RNases were purified from nitrogen-starved fission yeast cells in which cellular RNA was being degraded drastically. The two RNases showed similar properties. Their molecular weight in native form was about 170kDa. They were endoRNases which required divalent cations. PMID- 8247737 TI - Synthesis of oligonucleotides carrying linker groups at 1'-position of sugar residues. AB - A novel 2'-deoxyuridine analogue carrying an aminobutyl linker at 1'-position of the sugar residue was synthesized and incorporated into oligonucleotides, then an anthraquinone derivative was attached to the amino group. The oligonucleotides carrying the linker groups were more resistant to nuclease P1 and venom phosphodiesterase than an unmodified oligonucleotide. PMID- 8247738 TI - Enzymology and gene technology of Streptomyces nucleotide 3'-pyrophosphokinase 2',3'-cyclic monophosphokinase (PPKase). AB - Streptomyces extracellular nucleotide 3'-pyrophosphokinase-2',3'-cyclic monophosphokinase transfers 5'-beta,gamma-pyrophosphate group from ATP, some ATP derivatives and dATP to a variety of nucleot(s)ides at the 3'-OH site and synthesizes the respective 3'-pyrophosphoryl nucleotides. The enzyme also utilizes A-5'-pn (n = 3-5)-5'-N, transferring their adenosine 5'-pyrophosphoryl group and concomitantly eliminating the AMP moiety therefrom, leading to the synthesis of 2',3'-cyclic monophosphoryl nucleotides. The enzyme gene and its neighbouring up- and downstream sequences were analyzed. Streptomyces and enteric bacteria E. coli and Klebsiella pneumoniae transformants were constructed by incorporation of the pyrophosphokinase gene-recombined expression plasmids. Various effects of the treatments--intracellular synthesis of 3'-pyrophosphoryl nucleotides, retarded cellular growth, stimulated N2 fixation by K. pneumoniae and so on--were seen. PMID- 8247739 TI - Misincorporation of ribonucleotides by DNA polymerase during in vitro DNA replication. AB - The fidelity of DNA replication with respect to misincorporation of ribonucleotides into DNA has been studied. Primed M13 DNA was replicated by E. coli DNA polymerase I in the presence of four dNTPs, a single rNTP and Mg2+ as a cofactor. All four rNTPs were incorporated into DNA in place of the corresponding dNTPs when rNTP/dNTP ratio was 1000:1. The frequency of rNTP misincorporation was C > G > A > U. PMID- 8247740 TI - Singly-linked catenation and knotting of cisplatin-DNA adduct by DNA topoisomerase I. AB - We studied the topological interaction between topoisomerase I (topo I) and cisplatin-DNA adduct. The topoisomers of the cisplatinized DNA formed by topo I are different in none-, low- and high-cisplatin concentrations to bind to DNA. Two unique topologically distinct invariants, singly-linked catenane and torefoiled knot DNA generated from cisplatin adduct, by topological reaction but not from none cisplatinized DNA. The results are discussed in relation with a possible recombinational role of cisplatin on the topological reaction with DNA. PMID- 8247741 TI - Cloning of a gene required for the alkaliphily of alkaliphilic Bacillus sp. strain C-125. AB - An alkali-sensitive mutant was obtained from alkaliphilic Bacillus sp. strain C 125. A DNA fragment required for the alkaliphily was cloned from the chromosomal DNA of strain C-125 by using the alkali-sensitive mutant as a host. The nucleotide sequence of the cloned DNA fragment indicated the occurrence of several ORFs. Comparison of these ORFs with sequences in the available databases revealed no closely related sequences. PMID- 8247742 TI - Cloning and structural analysis of a human thymidylate synthase pseudogene splitted by several Alu sequences. AB - An unidentified genomic DNA fragment of 2.4kb that is weakly hybridizable with thymidylate synthase (TS) cDNA was cloned from a human genomic DNA library. Sequencing of the cloned DNA fragment and comparison of the sequence with that of the known human TS cDNA revealed that the DNA fragment contained a human TS processed pseudogene with unusual features. Based on the rate of nucleotide substitutions for neutral mutations in the 3'-untranslated regions between the gene and the pseudogene, it was estimated that the human TS pseudogene was formed about 16 million years ago. PMID- 8247743 TI - Synthesis of full-length cDNA using DNA-capped mRNA. AB - We have developed a novel method for synthesizing a full-length cDNA based on the addition of DNA-RNA chimeric linker to the CAP site of mRNA. The DNA-capped mRNA was used as a template for cDNA synthesis with a dT-tailed vector primer. The model experiment using rabbit globin mRNA showed that more than 80% of the clones encoding alpha- and beta-globin contains a full-length cDNAs starting from the CAP site. PMID- 8247744 TI - Construction of the Bacillus subtilis chromosome physical map and the strategy for mapping newly isolated genes in one membrane filter for hybridization. AB - A complete physical map of the Bacillus subtilis 168 chromosome was constructed. The merging of this physical map is expected not only to provide important insights into the organization and rearrangement of genes of this species but also to be a powerful means for the genome analysis. One of the most practical aspects is rapid and accurate mapping of newly isolated genes using a single membrane filter for hybridization. This protocol proved that not only unique genes but also multiple homologous genes dispersed on the chromosome can be physically mapped. PMID- 8247745 TI - Controlling the conformation of large DNA molecule in aqueous solution. AB - We have studied the conformational change of T4DNA molecules in aqueous polyethylene glycol (PEG) solution by use of fluorescence microscopy. It was found that the DNA molecules exhibit abrupt transition from elongated coil into shrunken globule conformations depending on the concentration of PEG6000, a polymer with 186 monomer units. Interestingly, PEG20M, with 454 monomer units, shows two different types of transition; coil-globule transition around 0.1 M and globule-coil transition above 1 M. Such transitions are expected to be related to the regulation of gene expression and replication in living cells. PMID- 8247746 TI - The hydrophobic and electrostatic effect of basic polyamino acid-DNA polyion complex on artificial bilayer lipid membrane. AB - Interaction between DNA-polyion complex, such as DNA-poly-L-arginine complex and DNA-histon complex, and artificial bilayer lipid membrane was studied by measuring the transmembrane current fluctuation. DNA did not interact with neutral bilayer lipid membrane. However, with the application of a positive potential, a large current fluctuation was observed in the presence of DNA-poly-L arginine complex. Using a negative potential, such a current fluctuation was not observed. This current fluctuation was dependent on the polarity of the applied potential. On the other hand, during the application of both positive and negative potential, large current fluctuation was observed in the presence of the DNA-histon complex. These results indicate that DNA-histon complex is able to permeate across the membrane and DNA-poly-L-arginine complex is able to be adsorbed on the hydrophobic part of membrane. PMID- 8247747 TI - Meso-oligodeoxyribonucleotides. AB - Meso-DNAs, containing L-sugar and D-sugar in an alternate sequence as the backbone [Fig. 1, designated as LD-(dN)n], were prepared as a modificated oligonucleotides of natural- and enantio-DNA. The characteristics of meso-DNAs were analyzed, focusing on LD-(dA)n and LD-(dT)n. These L-sugar containing oligomers show resistancy to phosphodiesterases. Though both LD-(dA)n and LD (dT)n bound their corresponding complementary homopolymers, apparent differences of the interaction mode between these two meso-DNAs were observed: [1] Concerning the complex formation with the complementary homopolymers, LD-(dA)n favors a triplex formation while LD-(dT)n forms only duplex, and [2] Concerning DNA/RNA selectivity for the complex formation, LD-(dA)n interacted with poly U more strongly than poly dT (RNA-selective), while LD-(dT)n showed stronger interaction with poly dA than poly A (DNA-selective). The similarities in CD-spectra of LD oligomer/natural polymer complexes to natural complexes suggest that the conformation possesses a structure close to a natural right-handed helix conformation. PMID- 8247748 TI - A possible function of DNA curvature in transcription. AB - The region preceding the beta-lactamase promoter of Escherichia coli plasmid pUC19 has a DNA curvature (curved DNA or bent DNA). Studies using mutant plasmids showed that the curved DNA structure is required for efficient transcription of the beta-lactamase gene and suggested that the requisite shape is a small part of a right-handed coil. Based on these results, the possible functional significance of DNA curvature in transcription will be discussed. PMID- 8247749 TI - The changes of c-fos mRNA expression in the protein kinase C activated osteoblastic MC3T3-E1 cells cultured in a low-calcium environment. AB - The effect of phorbol 12-myristate 13-acetate (TPA) on the c-fos mRNA expression in the osteoblastic MC3T3-E1 cells cultured in a low-calcium environment was examined. The c-fos mRNA expression in MC3T3-E1 cells with the treatment of TPA was enhanced under the low-calcium environment. This fact suggests that TPA induced-protein kinase C enhancement produced the increase of c-fos mRNA expression, and that protein kinase C may be intimately involved in c-fos mRNA expression. PMID- 8247750 TI - Group I self-splicing introns in both large and small subunit rRNA genes of Chlorella. AB - We report the presence of Group I self-splicing introns in both nuclear small subunit (SSU) and large subunit (LSU) rRNAs of the unicellular green alga Chlorella ellipsoidea. The SSU intron (442 nt) was located at a position within the highly conserved helix 48 that was close to the 3' terminus of SSU rRNA: the position was exactly the same as previously reported for the Pneumocystis carinii Group I intron. The LSU intron (445 nt) was found in a highly conserved helix region (between positions 924-925) of LSU rRNA. Both introns can be folded into the secondary structures characteristic of Group I introns. The Chlorella introns self-spliced in vitro, yielding the typical Group-I intron splicing intermediates and products. Unlike the ordinary introns, however, splicing of the SSU intron was inhibited by high concentrations of monovalent cations at the second stage of splicing reaction. The Chlorella introns offer an opportunity to study the origin and evolution, physiological roles, and relationship between structure and splicing activity of Group I self-splicing introns. PMID- 8247751 TI - Quantification analysis of splice signal sequences. Exon skipping in beta-globin gene of thalassemia. AB - Nucleotide sequences of human beta-globin and its mutant genes were analyzed by quantification method. In the mutant, a GT-->AT change occurs at the 5'-splice site of the second intron, causing skipping of the whole second exon during RNA splicing. A mechanism of exon skipping was proposed on the basis of the intensity of 3'-splice signal sequence. PMID- 8247752 TI - Ribosomal RNA identity elements for recognition by ricin and by alpha-sarcin: mutation in the putative CG pair that closes a GAGA tetraloop. AB - Alpha sarcin is a ribonuclease that cleaves the phosphodiester bond on the 3' side of G4325 in 28S rRNA; ricin A-chain is a RNA N-glycosidase that depurinates the 5' adjacent A4324. These single covalent modifications inactivate the ribosomes. An oligoribonucleotide that reproduces the structure of the sarcin/ricin domain in 28S rRNA was synthesized and mutations were constructed in the 3'C and the 5'G that surround the GAGA tetrad that has the site of toxin action. Analysis indicates that catalysis by ricin requires a Watson-Crick pair to shut off a putative GAGA tetraloop, whereas, alpha sarcin does not. One interpretation is that there are alternate conformations of the sarcin/ricin domain in 28S rRNA and that one of the conformers is recognized by sarcin and the other by ricin A-chain. This switch in the structure could underlie the translocation of peptidyl-tRNA from the A to the P site and the vectoral displacement of mRNA one codon during elongation. PMID- 8247753 TI - Stability of the TAR RNA and its complex with arginine. AB - Most stable secondary structures and their stabilization energies of the TAR RNA with +1 to +104 nucleotide-sequence region were calculated at different temperatures by using thermodynamic parameters for RNA structure prediction. The most stable secondary structure has one bulge and one loop within the region of +20 to +40 nucleotide sequence, and its stabilization energy at 37 degrees C was 46.3 kcal mol-1. The interaction of a TAR bulge oligomer (TARBO) with arginine (Arg) which was in the binding site of a Tat protein was also investigated by CD measurements. The addition of Arg did not affect the CD spectrum of TARBO. The result was different from that of the RNA oligomer with both bulge and loop. PMID- 8247754 TI - DNA binding properties of c-Myc-related bHLH/LZ oncoproteins. AB - Structural features and DNA binding properties of bacterially expressed c-Myc related oncoproteins were examined. The circular dichroism spectra demonstrated highly alpha-helical nature of these proteins. Dimerization and the DNA binding of these proteins were examined by electrophoretic mobility shift assay. PMID- 8247755 TI - Mechanism of DNA triplex formation and its specificity as studied by filter binding assay. AB - The filter binding method was found to be a very useful method for triple-helical formation of oligo DNA duplexes with homoprymidine single-strands. Using this method, we have obtained dissociation constants of triplexes and association rates of triple-helical formation of a variety of combinations of double-strands (23-mer) and pyrimidine single-strands as functions of pH and temperature. pH dependences of dissociation constants and association rates are theoretically discussed in terms of acid-base equilibrium of pyrimidine strands. Temperature dependence of dissociation constants and association rates were considerably different between acidic and neutral pH range, suggesting that mechanism of triple-helical formation differs between two pH ranges. The results were best interpreted in terms of a three-state model for the triple-helical formation. Furthermore, the effect of mismatched sequences on the stability of the triplexes were also discussed. PMID- 8247756 TI - Preliminary crystallographic studies on hammerhead ribozymes. AB - Crystallization of hammerhead ribozymes was tried by the hanging drop vapor diffusion method. The crystals grown up to 0.5 x 0.05 x 0.05 mm3 in size gave diffraction patterns with about 5 A resolution using synchrotron radiation. The crystal was determined to be trigonal with space group P3(1) or P3(2) and cell parameters a = b = 49.6 A and c = 53.3 A. PMID- 8247757 TI - Importance of specific purine-pyrimidine amino and hydroxyl groups for efficient cleavage by a hammerhead ribozyme. AB - Subsequent of the discovery that RNA can have site specific cleavage activity, there has been a great deal of interest in the design and testing of trans-acting catalytic RNAs as therapeutic agents. We have chemically synthesized and experimentally analyzed chimeric DNA-RNA ribozymes in which the catalytic center remains as RNA. PMID- 8247758 TI - Cleavage of the highly conserved hairpin-loop region of HIV-1 by synthetic ribozymes. AB - Hammerhead ribozymes can be considered as molecular scissors for suppressing AIDS virus. It is attractive to target single-stranded region of HIV-1 RNA because single-stranded RNA interactions or "kissing" induce hybridization. We, therefore, chose a highly conserved hairpin-loop structure, that is located in the 5' long terminal repeat region and just downstream of the TAR regulatory sequence. As a model system, we synthesized the corresponding hairpin-loop structure (21-mer RNA) and several types of ribozyme that are different in sizes in the hybridizing arms. These ribozymes successfully cleaved the 21-mer hairpin structured substrate after CUU sequence but with different efficiencies. PMID- 8247759 TI - Theoretical analyses on the role of metal cations in RNA cleavage processes. AB - Previously, we found that locations of metal cations in biomolecules, such as proteins and nucleotides, were in accord with predictions based on electrostatic potential calculations of the nearest polar functional groups. Electrostatic potential of a cyclic oxyphosphorane dianion 2, which is a model compound for an intermediate or a transition state for the hammerhead ribozyme reaction, indicates that Mg2+ coordinations at the regions between axial (bridging) and equatorial (phosphoryl) oxygens are most favorable. If we consider the fact that negative charge is concentrated on two of the equatorial phosphoryl oxygens in the transition state 2, the axial-equatorial coordination is an unexpected result. Nevertheless, experimental results also support such an axial-equatorial coordination. Therefore, hammerhead ribozyme is a metalloenzyme in which at least one Mg2+ ion acts as a base and possibly a second Mg2+ ion acts as a Lewis acid. The double Mg2+ ion catalysis may be a common mechanism among various types of ribozyme and some protein enzymes as well. PMID- 8247760 TI - Structural analyses of synthetic immobile Holliday junction. AB - In order to study the structural properties of Holliday junctions, we designed and synthesized oligodeoxyribonucleotides to construct the models of the Holliday junctions with no sequence symmetry at the sites of branching called immobile junctions. Electrophoretic and spectroscopic analyses of the immobile junctions indicated that the Holliday junctions constructed from four deoxy 24-mers and four 18-mers were stable in 0.1 M NaCl at approximately 5 x 10(-6) M strand concentration at room temperature. Addition of MgCl2 to the solution of HJs resulted significantly in an increase in melting temperature of HJs. PMID- 8247761 TI - Precise analyses of DNA structure by NMR. AB - Novel 1H NMR techniques were developed and applied to the analyses of the DNA structure. They distinguished the base proton signals of cytosine (uracil) from those of the other bases. Two dimensional experiments were also performed by using these techniques, and were found to be useful for signal assignments. Moreover, the (2'R)-[2'-2H]-labeled DNA 10-mer and 17-mer were synthesized to be used for the determination of precise structures. These stereoselective [2'-2H] labeling made possible explicit stereospecific assignments and exact determination of the vicinal coupling constants, 3JHH, and thus the conformation of each deoxyribose ring was accurately determined. PMID- 8247762 TI - Model building program HLX4: application to non-classical helical structures. AB - The model building program HLX4 for multiple stranded helices was developed as a useful tool for understanding of the biological roles of these helices and application of nobel base pairing scheme by chemical modification. In stead of torsion angle, the helical parameter is elected as the input information to generate the starting global structure. The model building performance is threefold, first, coarse connectivity check and second, fine connectivity check and energy minimization. This program system can be successfully applied to the DNA duplex with the nobel G:A base pairing and triplex with the modified sugar moiety. PMID- 8247763 TI - Thermodynamics of tetraplex DNAs, (T2G4)n and (T4G4)n. AB - The ends of eukaryotic chromosomes, termed telomeres, contain stretches of tandemly repeated guanine-rich sequences, such as (T2G4)n and (T4G4)n, along one strand. These sequences can form defined folded tetraplex structures in solution. Here we have systematically investigated the thermodynamic properties of a series of sequences, Tet n: (T2G4)n and Oxy n: (T4G4)n (n = 1, 2, 3, 4), in 10mM NaPi, 250mM NaCl, pH7.0, using differential scanning calorimetry (DSC). The melting process of all tetraplex DNAs is reversible. Tet n and Oxy n are similar in the dependence of melting temperature, Tm, and transition calorimetric enthalpy per strand, delta Hcal, on the number of the tandem sequence. The total delta Hcal value of Tet n is similar in magnitude to that of Oxy n, whereas the Tm value of the main fraction of Tet n is greater than that of Oxy n. These results suggest that Tet n and Oxy n form similar tetraplex DNA configuration, and the difference in Tm values between Tet n and Oxy n is attributed to the length of the T strings. PMID- 8247764 TI - Synthesis of cholesteryl supports and phosphoramidites containing a novel peptidyl linker for automated synthesis of triple-helix forming oligonucleotides (TFOs). AB - Preparation of G-rich TFOs containing cholesterol at the end presents challenging problems due to low yield in synthesis and loss in purification due to aggregation. These compounds also have low solubility which can result in variability in their uptake properties and cellular efficacy. To overcome these problems we have designed and synthesized cholesteryl -CPG (6) and -TentaGel (7) supports and phosphoramidite (8) containing a triglycyl linker. PMID- 8247765 TI - Loop-like structure of dGTTG in solution. AB - A strong NOESY cross peak was observed between G1H8 and G4H8 for a single stranded tetradeoxyoligonucleotide dGTTG in aqueous solution. If dGTTG is in an expanded form, NOE should not be observed because the distance between G1H8 and G4H8 is estimated to be about 13 A. This NOESY cross peak shows that the guanine residues of the both ends of dGTTG exist with short distance (< 5 A), and that dGTTG has a distorted loop-like structure in aqueous solution. This is the shortest single-stranded oligodeoxynucleotide that has a rigid structure in solution. PMID- 8247766 TI - Tertiary structural analysis of Escherichia coli lysine tRNA. AB - It has been suggested that Escherichia coli lysine tRNA (E. coli tRNA(Lys)) has an unusual anticodon loop structure, judging from the abnormal CD pattern derived from 5-methylaminomethyl-2-thiouridine (mnm5s2U) located at the first position of the anticodon of the tRNA. Here, the structure of E. coli tRNA(Lys) was analysed by chemical modification and nuclear magnetic resonance (NMR) of the imino protons. The chemical probing approach showed that the anticodon arm region forms the ordinary stem-and-loop structure as expected from the standard clover-leaf structure, because N7 of G30, N3 of C40, N7 of A41 and N7 of A42 were protected under a native condition. By using the heteronuclear 15N-1H single quantum coherence (HSQC) spectra of 15N-labeled E.coli tRNA(Lys) and the analyses of the nuclear Overhauser effects (NOE) between the imino protons of non labeled E.coli tRNA(Lys), the imino protons in the acceptor stem was assigned, which indicates that the acceptor stem exists in an usual form. The tertiary base-base interactions was also detected. Thus, E.coli tRNA(Lys) seems to maintain a similar structure to the ordinary L-shaped tertiary structure, as a whole. The fine structure of the anticodon loop is now being analyzed. PMID- 8247767 TI - Molecular design of artificial hydrolytic nucleases and ribonucleases. AB - Artificial nucleases and ribonucleases, which selectively hydrolyze the specific phosphodiester linkages in DNAs and RNAs, are prepared by the attachment of organic and inorganic catalysts to DNA oligomers as sequence-recognizing moieties. A hybrid of ethylenediamine and a 19-mer DNA selectively hydrolyzes tRNA(Phe) at the 3'-side of C63. Site-selective hydrolysis of DNAs has been accomplished by the lanthanide complexes bound to DNA oligomers. No concurrent oxidative cleavage of the ribose or the deoxyribose takes place. PMID- 8247768 TI - Structural basis for recognition of DNA by zinc finger and POU domain transcription factors. PMID- 8247769 TI - Solution structures of Myb DNA-binding domain and its complex with DNA. AB - The DNA-binding domain of Myb consists of three imperfect tandem repeats, each of which contains three conserved tryptophans. The solution structure of the third repeat was established by NMR; it includes three helices maintained by a hydrophobic core formed by three tryptophans, together with two histidines, and the second and third helices form a helix-turn-helix-related motif. Here, we report the whole structure of the DNA-binding domain, as determined by NMR. Each of the first and second repeats resembles the third one in structure, retaining three helices. As a whole, the DNA-binding domain contains an entirely unique structure, a triple helix-turn-helix-related motif. To determine the role of each repeat in DNA-binding, a 16-mer DNA duplex that is specifically recognized by Myb was synthesized and then its complex with the DNA-binding domain has been investigated by 3D-NMR. The interaction mode between the whole DNA-binding domain and the duplex has shown that the third helix in each repeat likely locates in the majour groove of DNA. It is suggested that the Myb DNA-binding domain wraps around the DNA duplex by fitting the triply repeated helix-turn-helix-related motifs in the majour groove. PMID- 8247770 TI - Structural features and properties of an extraordinarily stable hairpin-turn structure of d(GCGAAGC). AB - The extraordinarily stable hairpin-like structure formed by a short DNA fragment, d(GCGAAGC), has been studied by NMR spectroscopy and UV melting behaviour. The fragment is folded back between A4 and A5, and forms two terminal G-C base pairs and a non-Watson-Crick G-A base pair. All the nucleotides adopt C2'-endo sugar puckers. Both G1C2G3A4 and A5G6C7 moieties have characteristics of B-form geometry and within each moiety all the bases are involved in extensive base-base interactions. Distortion from B-form occurs in only the three torsion angles between A4 and A5 residues, which causes a sharp turn in the structure. There is no clear distinction between a stem and a loop region, as observed in usual hairpin structures, so that we classify it as a turn (hairpin-turn) structure. In addition to the thermal stability, this fragment is more stable towards the attack of some nucleases than other single-stranded as well as usual hairpin DNA fragments. PMID- 8247771 TI - Tertiary structures of mitochondrial tRNAs having characteristic secondary structures. AB - Some of the animal mitochondrial (mt) tRNAs are thought to have unusual secondary structures as inferred from the mt DNA sequence analysis. To obtain information on the relationship between the unusual secondary and tertiary structures and function of these mt tRNAs at the RNA level, we have analyzed the structure of bovine mt serine tRNAs and obtained the following results. 1) Using the novel secondary structure proposed on the basis of enzymatic probing and phylogenetic comparison and the known tertiary structure of yeast tRNA(Phe), a tertiary structural model of tRNA(Ser) specific for UCU/C/A/G codons was constructed by a computer modeling. In the proposed model, one-base-pair elongation of anticodon stem compensated for the deletions in some of the loop regions of this tRNA, and the model maintained the topological relationship between the anticodon and 3' CCA terminus as nearly the same as that of yeast tRNA(Phe). The results of chemical modification of this tRNA supported the proposed model. 2) For NMR analysis of tRNA(Ser) specific for AGU/C codons and lacking the D stem, a large scale preparation of the unmodified tRNA transcript has been performed by the use of an in vitro T7 transcription system. PMID- 8247772 TI - Mechanistic aspects of the interaction of antisense DNA. AB - To investigate the mechanism of antisense effects, diastereoisomerism of oligodeoxynucleotides with modified internucleotide phosphate linkages such as oligodeoxynucleoside phosphoramidates (OPA) and oligodeoxynucleoside phosphorothioates (OPT). Reversed-phase HPLC was used to separate the diastereoisomers of OPA (up to tetramer) and OPT (up to pentamer), and it was found that the complete purification of all diastereoisomers could be achieved by the two-step chromatographic approach. From the circular dichroism measurements, it was found that some diastereoisomers formed certain hybrids with their complementary oligonucleotides and some did not. PMID- 8247773 TI - Oligodeoxynucleotides containing 2'-fluoro-arabino-nucleotide and C-nucleotide. AB - Oligonucleotides containing 2'-fluoro-ara-pyrimidines and 1-methyl-2'-deoxy-psi uridine were synthesized. The melting profiles show that the presence of FMAU has a large stabilizing effect on the duplex. Replacement of dT with FAU or psi-dT, or dC with FAC resulted in the formation of less stable duplexes. Temperature dependent CD spectroscopy demonstrated that the structures of these oligomers are very similar to those of unmodified oligomers, however, these minor alterations in structure of dodecamers resulted in faster splitting by endonuclease EcoR1. PMID- 8247774 TI - Nucleotides bearing a cleavable genotoxic group on the phosphate. AB - A new class of nucleotide derivatives in which N-nitroso-pyrrolidine (NPYR) is linked to the phosphate group through an ester linkage at the alpha-carbon of the nitrosamine were prepared by a reaction between a nucleoside monophosphate and alpha-acetoxy-NPYR. The NPYR derivatives were prepared from dpT, dpC, dpA, dpG, pA and dpCpT. Treatment of the NPYR-pX with acid, snake venom phosphodiesterase, or near ultraviolet light caused cleavage of the NPYR group, regenerating pX. All of these nucleotide derivatives were directly mutagenic towards Salmonella typhimurium TA1535. When phage M13mp2 double-stranded covalently closed circular DNA was treated with NPYR-dpT with near ultraviolet irradiation, single strand breaks of the DNA took place. PMID- 8247775 TI - Signal transduction in two light-transducing systems: bacteriorhodopsin and mammalian rhodopsin. PMID- 8247776 TI - Chemical characteristics of complexes of O6,9-dimethylguanine with cis-platin and debenzylation rates of O6-benzylguanines. AB - Complexes of O6,9-dimethylguanine with cisplatin were synthesized and their chemical characteristics were studied in relation to the function of the repair enzyme, O6-alkylguanine-DNA alkyltransferase (AGAT). In addition, dealkylation rates of O6-benzylguanine derivatives, that are though to be better substrates for AGAT, were determined by the thiophenol-Et3N system. PMID- 8247777 TI - Chemical characteristics of N-aminated nucleic acid bases. Nitrosation of aromatic N-amino group and accompanying reductive deamination. AB - Primary NH2 group attached to the pyridine-type or pyrrole-type nitrogen underwent sequential nitrosation and reductive deamination by the treatment with NaNO2-AcOH. Secondary NHR group attached to the pyridine-type nitrogen underwent also the reductive deamination. In contrast, secondary NHR group attached to the pyrrole-type nitrogen gave the corresponding N-nitroso derivative under the same conditions. Mechanism of the reductive deamination is discussed. PMID- 8247778 TI - Stereocontrolled synthesis of beta-2'-deoxy and 2',3'-dideoxynucleosides by intramolecular glycosylation. AB - Intramolecular glycosylation of phenyl 3-O-benzyl-2-deoxy-5-O-(4-methoxy-2 pyrimidyl)-1-thio-D-ribofuranoside and 2,3-dideoxy-5-O-(4-methoxy-2-pyrimidyl)-1 thio-D-glycero-pentofuranos ide by activation with dimethyl(methylthio)-sulfonium tetrafluoroborate followed by hydrolysis gave the corresponding beta-2'-deoxy and beta-2',3'-dideoxypyrimidine nucleoside derivatives respectively in good yields. PMID- 8247779 TI - The synthesis of alternating alpha,beta-oligodeoxyribonucleotides with alternating (3'-->3')- and (5'-->5')-internucleotic linkages as potential therapeutic agents. AB - A simplified synthesis of alpha-2'-deoxyribonucleoside derivatives has been developed and the solid-phase preparation of antisense alpha,beta oligodeoxyribonucleotides with alternating (3'-->3')- and (5'-->5') internucleotidic phosphodiester linkages, targeted against HIV-1 tat mRNA, has been accomplished. Hybridization properties of these oligonucleotide analogues have been investigated. PMID- 8247780 TI - Stereo-controlled C-C bond formation at the anomeric position of uracil nucleosides. AB - Stereoselective synthesis of 1'-carbon-substituted uracil nucleosides has been achieved through face-selective bromo-pivaloyloxylation of a 1',2'-unsaturated derivative and successive SnCl4-promoted nucleophilic substitution with organosilicon reagents. This constitutes the first example of C-C bond formation at the anomeric position of nucleosides. PMID- 8247781 TI - Synthesis and biological activity of 1-(2-deoxy-2-C-fluoromethyl- and 2-C hydroxymethylarabinofuranosyl)-cytosines. AB - We newly synthesized 1-(2-deoxy-2-C-fluoromethyl- and 2-C-hydroxymethyl arabinofuranosyl)cytosines and evaluated their biological activities. The syntheses of these compounds were achieved by radical deoxygenation of tert alcohol of 2'-position of the corresponding fluorohydrine and acetoxymethyl derivative. 1-(2-Deoxy-2-C-fluoromethylarabinofuranosyl)cytosine (5) showed potent antileukemic and anticytomegalovirus activities. PMID- 8247782 TI - Convenient and large scale synthesis of the cap structure, m7G5'pppG. PMID- 8247783 TI - Synthesis of 4'-methyl-4'-thiothymidine. PMID- 8247784 TI - Highly stereoselective synthesis of bicyclic-sugar adenosine derivatives via Diels-Alder reaction. PMID- 8247785 TI - Synthesis and biological activity of 2-substituted analogues of 2'-deoxy-2' fluoroadenosine. AB - Reaction of 2-iodo-6-methoxypurine 2'-deoxy-2'-fluororiboside with ammonia in methanol gave 2-iodo-adenosine derivative 2 in 69% yield. Treatment of 2 with nucleophiles gave 2-substituted analogues of 2'-deoxy-2'-fluoroadenosine. A new 2'-deoxy-2,2'-difluoroadenosine was also prepared by way of quaternary salt 4. PMID- 8247786 TI - Effects of substituents on the acid-catalyzed photoreaction of pyrimidine derivatives in p-xylene. AB - In order to obtain insight into the protonation of the electron adducts of pyrimidine bases (I) and the chemical behavior of the resulting radicals under an acidic condition, the photoreaction of 5-substituted 1,3-dimethyluracil (1) (substituents; p-xylyl, methyl, H, Cl, and F) was performed in p-xylene (2) in the presence of a large amount of trifluoroacetic acid (TFA). 5, 6-dihydrouracil derivatives (3), 5-p-methylbenzyluracils (4) and the 6-isomers (5) were yielded as the major products. Substitution of an electron-releasing group (p-xylyl) at C 5 raises the ratio of the products from the C6-protonated pyrimidine electron adduct (III) (3, 4) vs. the adduct from the O4-protonated isomer (II) (5), while the electron-withdrawing substituents (F > Cl) suppress the ratios. PMID- 8247787 TI - Design and synthesis of regioselective cleaving reagents for oligonucleotides. AB - Reaction of methyl 2,3-anhydro-alpha-D-ribofuranoside with allylmagnesium chloride selectively took place at the 2-C position to afford methyl 2-C-allyl-2 deoxy-alpha-D-arabinofuranoside. 1-O-Acetyl-2,3,5-tri-O-benzoyl-beta-D ribofuranose reacted with allylic alcohols in the presence of BF3.OEt2 to give the corresponding allyl beta-D-ribofuranosides. The allyl group in these compounds were converted into a variety of functional groups which include bromoacetoxy, oxylanyl, tosyloxy, and azide. PMID- 8247788 TI - Synthesis of 1-(5-deoxy-5-fluorohexofuranosyl)uracil analogs. PMID- 8247789 TI - Acyclic analogues of naplanocin A as a potential inhibitor of S adenosylhomocysteine hydrolase. PMID- 8247790 TI - Dialkylformamidine protected deoxyadenosine phosphoramidites in oligonucleotide synthesis. AB - A series of dialkylformamidine protected deoxyadenosine phosphoramidites were prepared for automated, solid-support DNA synthesis. The set of Abz, Gdmf, Cbz, T phosphoramidites gave high purity and high yield oligonucleotides, with rapid and complete deprotection at 65 degrees C in one hour. Different times and temperatures of exposure to concentrated ammonium hydroxide were examined to establish the optimum conditions for deprotection of oligonucleotides. PMID- 8247791 TI - Sedative and hypnotic activities of pyrimidine nucleoside derivatives. AB - N3-Substituted pyrimidine nucleoside and their related compounds were synthesized. The central nervous system (CNS) depressant effect of the compounds such as hypnotic activity and synergistic effect with pentobarbital have been evaluated. N3-Phenacyl derivatives of uridine (2), thymidine (3), deoxyuridine (4), 6-azauridine (5) and arabinofuranosyluracil (6) exhibited the hypnotic activity, whereas N3-phenacyl-2',3'-O-isopropylideneuridine did not. Compound 2, 3, 4 and 6 significantly enhanced pentobarbital-induced sleep. The results indicate that CNS depressant effect of pyrimidine nucleoside derivatives might relate to functional group at the N3-position on oxopyrimidine ring and stereospecificity of sugar moiety at the N1-position. PMID- 8247792 TI - Enzyme aided regio-selective acylation and deacylation of nucleosides. AB - Selective protection and deprotection of 5-fluorouridine and 5-fluoro-2'-deoxy uridine derivatives have been studied by enzymatic esterification in organic solvents and enzymatic hydrolysis. Different selectively protected products could be prepared by using different enzymes. This provides an efficient method for selective protection of nucleosides. PMID- 8247793 TI - Complexation of nucleotides by flexible molecular tweezers using pi-interaction and electrostatic interaction. PMID- 8247794 TI - The manufacture and molecular characterization of phosphorothioate deoxynucleotides for use as antisense therapeutics in human clinical trials. PMID- 8247795 TI - Biologically active oligodeoxyribonucleotides--I: Syntheses and anti-HIV-1 activities of 5'-modified pentadecadeoxyribonucleotides. AB - 5'-Modified pentadecadeoxyribonucleotides with a sequence (5'-TGGGAGGTGGGTCTG-3') (15mer) complimentary (antisense) to the tat 2nd splicing acceptor region of human immunodeficiency virus type 1 (HIV-1) were prepared and evaluated for anti HIV-1 activity. The modified antisense oligodeoxyribonucleotides (AS-ODNs) were synthesized using 5'-modified thymidine 3'-phosphoramidites as the 5'-terminal residues. The structures of the 5'-modified 15mers were confirmed by negative ion LSI mass spectroscopy, and the anti-HIV-1 activities were evaluated in vitro by the MTT assay using MT-4 cells. While the unmodified 15mer had no activity in our assay system, the 15mers bearing modifications with trityl-type substituents at the 5'-end showed high anti-HIV-1 activities. PMID- 8247796 TI - Oligonucleotides of carbocyclic 5'-nor 2'- and 3'-deoxyadenosine. AB - Oligonucleotides possessing a shortened internucleotide bond have been prepared using carbocyclic nucleosides lacking the 5'-methylene as the monomeric units. Specifically, the oligomers derived from 5'-nor carbocyclic 2'-deoxyadenosine and 3'-deoxyadenosine will be described. PMID- 8247797 TI - Spectroscopic characterization of heterochiral DNAs. AB - We have synthesized heterochiral dodecadeoxynucleotides having an unnatural L nucleotide residue, and have investigated their structures by ultraviolet (UV) absorption, circular dichroism (CD) measurements and nuclear magnetic resonance (NMR) spectroscopy. It was clearly shown that the overall structures of the heterochiral 12-mers are a right-handed B-conformation and the unnatural L nucleotide residue in the heterochiral 12-mer (L-4) forms stable Watson-Crick type base-pairing with the natural complementary residue. In this double helix (L 4), the unnatural G4 residue has an S-type sugar conformer and a low-anti glycosidic torsion angle. From the properties of an enantiomer, natural nucleotides may possibly form a low-anti left-handed B-form duplex with the aid of certain factors. PMID- 8247798 TI - Synthesis of phenylphosphonodiester-modified oligodeoxyribonucleotides via the allyl protection. PMID- 8247799 TI - Double and triple helix formation of 2'-5' and 3'-5' oligonucleotides. AB - Double and triple helix formation of non-self complementary 2'-5' and 3'-5' linked oligonucleotides, 3'-5'(pA)7, 3'-5'(pU)7, 2'-5'(pA)7 and 2'-5'(pU)7, was studied using UV and CD spectroscopies. UV mixing curve and UV melting curve showed that 3'-5'(pA)7:3'-5' (pU)7 system could form double and triple helix, while 2'-5'(pA)7:2'-5'(pU)7 system could form only double helix and 3'-5'(pA)7:2' 5'(pU)7 system could form neither double nor triple helix under the condition examined. Thermodynamic parameters obtained from the UV melting curve indicate that the linkage type of oligonucleotides has profound effect on the helix formation. PMID- 8247800 TI - Non-radioisotope labelling of DNA with multifunctional labelling agent. AB - A novel bi-functional fluorescent labelling agent derived from 5-hydroxy-1 naphthalene sulfonic acid and tris(2-amino-ethyl)amine was prepared and incorporated to 5'-monophosphate function of DNA through phosphoramidate bond. The labelling agent was designed to bear fluorescent moiety as well as primary and tertiary amines which are expected to have positive charge at neutral pH range. A 15-mer DNA labelled with this reagent exhibited enhanced duplex forming ability with its complement compared to the corresponding un-labelled DNA. PMID- 8247801 TI - Sequence analysis of low-molecular-weight RNAs by the use of non-radioactive labeling. AB - A new procedure for non-radioactive labeling of the 3'-terminal -OH was developed to facilitate the sequencing works on low-molecular-weight RNAs. At first, biotinylated pCp (cytidine 3',5'-bisphosphate) was synthesized and ligated to the 3'-terminal of yeast tRNA(Phe) with the aid of RNA ligase. Although the labeling efficiency was high enough, this labeling resulted in a lack of the sequence ladders around the 3'-terminal 10 nucleotides. A longer oligonucleotide, p(dC)(dT)16-Biotin was designed to overcome this defect and proved to be a better non-radioactive 3'-labeling probe. PMID- 8247802 TI - Affinity gel electrophoresis of nucleic acids. The interaction between water soluble polymers having malachite green and double-stranded DNAs. AB - Copolymers prepared by the free radical copolymerization of acrylamide and vinyl malachite green were immobilized in an agarose gel matrix to produce a novel affinity gel for double-stranded DNAs. Relative electrophoretic mobilities of calf thymus DNA and poly(dA-dT)2 strongly depended on the amount of vinyl malachite green to show the strong interaction between DNAs and the copolymers. But any affinity interaction was not observed between poly(dG-dC)2 and the copolymers. PMID- 8247803 TI - Detection of mismatch positions on the DNA-polyvinyladenine hybrids using capillary affinity gel electrophoresis. PMID- 8247804 TI - Novel intercalators of pyrylium dye into double-stranded DNA. AB - Discovery is reported of novel intercalators without fused ring systems, pyrylium and thiopyrylium salt. 2-Methyl-4,6-bis-(N,N-dimethylaminophenyl) pyrylium salt 1 (Fig. 1) is, in particular, virtually non-fluorescent in aqueous solution, but form highly fluorescent complexes with double-stranded DNA (dsDNA), which yield more than 400 fold fluorescence enhancement at the peak emission. We report the synthesis, absorption and fluorescence properties of 1 and its thiopyrylium derivative 2 in comparison with other pyrylium 3, thiopyrylium 4 dyes and typical intercalator, ethidium bromide(EB). Compound 1 shows the absorption maximum at 540 nm and the red shift (about 40 nm) in the presence of dsDNA in similar to EB. The enhancement of fluorescence emission excited by visible light is extremely sensitive to the concentration of dsDNA comparing to that of EB. The thiopyrylium salt 2 with absorption maximum at 575 nm shows the similar profile in the absorption spectral shift and fluorescence enhancement to that of 1, when the dye binds to dsDNA. PMID- 8247805 TI - Equilibrium and kinetic studies of the interaction of the benzimidazole dye Hoechst 8208 with nucleic acids. AB - The interaction of the benzimidazole dye Hoechst 8208 (H8208) with nucleic acids has been studied by temperature-jump relaxation method. The dependence of relaxation times on the nucleic acid concentration indicates that H8208 binds to RNA by a single biomolecular reaction and to DNA by a bimolecular reaction, followed by a slow isomerization. PMID- 8247806 TI - The interaction of DNA with quinacrine and its analogs studied by absorption and circular dichroism spectroscopy. AB - The interaction of DNA with quinacrine and its analogs has been studied by absorption and circular dichroism (CD) spectroscopy. The results indicate that the 2-alkoxy group of the acridine ring plays an important role in the spectral behavior which depends on the base composition of DNA. PMID- 8247807 TI - Induced-fit association between DNA and esperamicin. Importance of structural flexibility in host DNA duplex. AB - In this study, a series of synthetic oligonucleotide duplexes are tested as a substrate for esperamicin. The duplexes contain a typical binding sequence of esperamicin, 5'-GGA/TCC, but have different flexibilities in helix structure from each other. When cleavage activities of these oligonucleotides by esperamicin were estimated by using DNA sequencing method, a substantial increase of the cleavage at 3'-NAGG was observed with increasing the helix flexibility. This observation indicates that structural flexibility of host DNA duplex is important in an induced-fit association between esperamicin and DNA. PMID- 8247808 TI - New strategies for oligonucleotide synthesis by use of 2-trimethylsilylethyl and 2-trimethylsilylethoxymethyl as the phosphate and 2'-hydroxyl protecting groups, respectively. AB - The 2-Trimethylsilylethyl (TSE) group was found to be useful as a protecting group for the internucleotidic phosphate in DNA synthesis. Oligodeoxyribonucleotides were synthesized in good yields by using phosphoramidite building blocks having the TSE group. In RNA synthesis, 2 trimethylsilylethoxymethyl (SEM) group was introduced as a new protecting group for the 2'-hydroxyl. Selective protection of this function with SEM-Cl was achieved in good yield by use of 3',5'-O-bis(tert-butyl)silanediyl ribonucleoside derivatives. PMID- 8247809 TI - DNA cleavage by benzidine derivatives in weak acidic conditions. AB - A plasmid DNA was cleaved by aromatic amines in weak acidic conditions at 37 degrees C. The cleavage was not inhibited by 4 hydroxylradical scavengers, EDTA, SOD and catalase. It was strongly influenced by the pH value of the reaction buffer. The decrease of Form I DNA in this reaction obeyed good first-order kinetics. PMID- 8247810 TI - Rational design for potentially anti-tumor and/or anti-viral nucleosides. PMID- 8247811 TI - Remarkable acceleration for the deamination of carbocyclic purine nucleosides by adenosine deaminase under high-pressure. AB - Deamination of an anti-HIV carbocyclic adenine nucleoside, 9-[c-4,t-5 bis(hydroxymethyl)cyclopent-2-en-r-1-yl]-9H-adenine [(+/-)-BCA] and (+/-)-diamino carbovir by adenosine deaminase under high-pressure (4 kbar) was carried out. Deamination of the former gave the corresponding (-)-hypoxanthine derivative whereas that of the latter produced (-)-carbovir having anti-HIV activity. The enantiomeric excess (e.e.) of both products was nearly 100%. PMID- 8247812 TI - Association of branched nucleic acids. AB - Various branched nucleic acids (bNA's) of the general formula, 5'-(N)xrA[2' (N)y], 3'-(N)y, have been prepared and their association with complementary linear oligonucleotides has been studied. The binding of complementary sequences to bNA's (for instance when N = dT, x = 0; y = 10 or x = y = 10 and the complement is dA10) yields transition temperatures (Tm's) equal to or greater than those of the corresponding linear complexes of similar base composition. Furthermore, a saturation of binding is observed when the number of mole equivalents of complementary sequence equals the number of "arms" less one (n-1), i.e., for the "Y"-shaped molecule, 5'-T10-rA[(2'-T10), 3'-T10]; n = 3, saturation occurs at two equivalents of complementary sequence. The significance of these results is discussed in terms of possible triple helix formation. PMID- 8247813 TI - Alternative answers. PMID- 8247814 TI - Effectiveness of pH measurements in predicting feeding tube placement: an update. AB - This paper reports further findings from an ongoing clinical study designed to evaluate the extent to which pH values of aspirates from feeding tubes can be used to differentiate between gastric and intestinal tube placement and gastric and respiratory tube placement. The sample consisted of 405 aspirates from small bore nasogastric tubes and 389 aspirates from nasointestinal tubes, which were obtained from 605 subjects ranging in age from 18 to 94 years. Data were collected at the time of initial placement and again, when possible, after feedings were initiated. A total of 794 pH-meter readings were made concurrently with X-rays to determine feeding tube position. Gastric placement was successfully distinguished from intestinal placement of the feeding tubes on the basis of pH-meter readings (p < .0001). Approximately 85% of the 405 pH-meter readings from gastric fluid were between 0 and 6.0, while over 87% of the 389 pH meter measurements performed on intestinal aspirates were greater than 6.0. Four aspirates from feeding tubes inadvertently placed in the respiratory tract (two in the pleural space and two in the tracheobronchial tree) were tested with a pH meter, all had pH values greater than 6.5. PMID- 8247815 TI - The effects of hydrogen peroxide rinses on the normal oral mucosa. AB - Oral mucosal effects of hydrogen peroxide mouth rinses were investigated in normal volunteers. Following a 2-week control period, 35 subjects were randomly assigned to rinse with either normal saline, 1/4-strength hydrogen peroxide (0.75%), or 1/2-strength hydrogen peroxide (1.5%), 4 times daily for 2 weeks. Mucosal status, buccal microbial adherence, salivary flow rate (SFR), and subjective reactions were assessed weekly. In the normal saline group, no significant changes were noted in any of the observed parameters and subjective reports were unremarkable. In both hydrogen peroxide groups, significant mucosal abnormalities were observed (p < 0.001) and subjective complaints were numerous. Bacterial adherence was significantly reduced in the 1/4 hydrogen peroxide group but not in the 1/2 hydrogen peroxide group. Despite reports of dry mouth, SFRs were not altered significantly. Since hydrogen peroxide rinses are associated with mucosal abnormalities and elicit overwhelmingly negative subjective reactions in normal individuals, they are not recommended for oral care. PMID- 8247816 TI - Cue familiarity reduces spatial disorientation following hippocampal damage. AB - The purpose of this study was to determine whether familiarity with a cue would enhance the ability of rats with hippocampus (HPC) damage to locate a cued hidden platform in the Morris water test. After preoperative training with the cue (n = 21) or handling only (n = 17), rats were given electrolytic bilateral HPC lesions or sham surgery for controls. All rats were then tested for 4 days, 6 trials per day, with the cue marking the platform location. Rats with HPC lesions familiar with the cue (FB) were significantly more efficient than rats with HPC lesions unfamiliar with the cue (UB) in swim time (M = 10.31 +/- 2.2 vs 46.72 +/- 7.5 sec, p < .05) and in directional heading error (M = 31.98 +/- 3.4 vs 57.92 +/- 4.0 degrees, p < .05) on Day 1. These differences occurred across the 4 test days and were shown to be the result of an impairment in learning of the cue task in the UB groups rather than simply an effect of more practice with the cue by FB animals. No significant differences were found between FB animals and familial controls. PMID- 8247817 TI - Temporal dissociation of recovery from antinociceptive and motor effects following intrathecal administration of a selective mu-opioid agonist. AB - In this series of studies, the antinociceptive and motor side effects associated with intrathecal administration of the selective mu-opioid agonist, DAMGO, were evaluated in the same rat. The Randall-Selitto paw-withdrawal test was used to measure mechanical nociceptive threshold, and an accelerating Rotarod treadmill was used to evaluate motor coordination. In the first experiment (N = 48), intrathecal administration of DAMGO produced dose-dependent increases in mechanical nociceptive thresholds that correlated with decreases in motor coordination. In the second experiment (N = 8), intrathecal administration of naloxone (5 micrograms) reversed both the antinociceptive effects and the motor deficits produced by 5 micrograms of intrathecal DAMGO. In the third experiment (N = 13), following administration of the highest dose of DAMGO (5 micrograms) as part of a cumulative dose response curve, paw-withdrawal and motor coordination testing were continued at 45-minute intervals for a period of 225 minutes. DAMGO induced deficits in motor coordination recovered significantly sooner than nociceptive responses. These data provide support for the suggestion that the motor as well as antinociceptive effects of spinal opioids are mediated through an action at an opioid receptor, but that these two effects may be mediated by different mechanisms. PMID- 8247818 TI - The effects of activity and bed rest on tissue oxygen tension, perfusion, and plasma volume. AB - An experimental study with a crossover design was used to compare the effects of activity and bed rest on subcutaneous oxygen (PscO2), perfusion (BFsc), and plasma volume, and to explore the relationship of cardiovascular fitness to these variables. Fifteen healthy males were randomized to bed rest or activity and later completed the remaining protocol. Exercise prescription for the activity protocol and cardiovascular fitness were based on a treadmill performance test. Repeated measurements were made of subcutaneous oxygen and temperature using an optode in upper arm subcutaneous tonometers. Perfusion and PscO2 were greater during bed rest (p < .05). Post hoc analysis revealed a trend toward higher heart rates and diastolic blood pressure during the activity protocol. Change in plasma volume did not differ between protocols, and cardiovascular fitness was unrelated to PscO2 or BFsc. Findings suggest that in uninjured individuals, higher levels of activity intensity reduce oxygen levels and blood flow in peripheral tissues. PMID- 8247819 TI - Inspiratory muscle training in patients with chronic obstructive pulmonary disease. AB - The effects of 6 months of muscle training with an inspiratory pressure load equal to 30% of the maximal inspiratory pressure (PImax) was compared with sham training with a light inspiratory pressure load. In this double-blind randomly assigned study, 67 patients with chronic obstructive pulmonary disease were followed for 6 months. Both groups demonstrated improvements in performance of PImax, respiratory muscle endurance time for breathing against an inspiratory pressure load equal to 66% of PImax, 12-minute distance walk, and dyspnea. The treatment group did not demonstrate significant effects beyond those observed in the control group. Improvements in performance were related to improved coordination of the inspiratory muscles and desensitization to dyspnea. PMID- 8247820 TI - Meperidine attenuates the febrile response to endotoxin and interleukin-1 alpha in rats. AB - Intravenous meperidine is commonly used to treat rigors and chills in febrile patients, though its mechanism of action is unknown. Therefore a laboratory model of pyrogen-induced fever was used to evaluate the effects of meperidine on the febrile response of rats injected with bacterial endotoxin or IL-1 alpha. Fever was measured using a computerized biotelemetry system for the continuous monitoring of body temperature. Injection of meperidine blocked the onset of fever in rats injected with endotoxin and attenuated the febrile response in rats injected with IL-1 beta. Furthermore, incubation of human mononuclear leukocytes (MNL) in the presence of meperidine significantly reduced endotoxin-induced secretion of IL-1 beta in vitro. These data suggest that meperidine decreases rigors and chills by decreasing the "set point" for fever, perhaps by reducing MNL secretion of IL-1 beta. PMID- 8247821 TI - Reliability of sleep diaries for assessment of sleep/wake patterns. AB - Although sleep diaries are widely used in clinical and research settings, only a few studies have compared the subjective information recorded in these diaries to objective information about sleep recorded. The goal of this study was to determine if a sleep diary could be used to obtain reliable data about home sleep/wake patterns over a 24-hour period. Fifty subjects (25 narcoleptic and 25 matched control subjects) completed a sleep diary while undergoing 24-hour ambulatory polysomnographic monitoring. The percentage agreement between the subjective data recorded in the sleep diaries and polysomnographic data was acceptable (kappa = .87). Sensitivity and specificity were also high (92.3% and 95.6%). The sleep diary is a reliable instrument for collecting data about sleep/wake patterns, but should be used with caution when collecting data from subjects who are likely to take frequent daytime naps. PMID- 8247822 TI - Testing the sensitivity of a biologic assay for mutagenicity. PMID- 8247823 TI - Re: 'Challenging misperceptions about nurses' moral reasoning'. PMID- 8247824 TI - Re: 'Issues in methods and measurement of thermodilution cardiac output'. PMID- 8247825 TI - Nursing Times open learning programme. M6: managing the process of providing care. Part (iv): Managing time (continuing education credit). PMID- 8247826 TI - Healthy living? PMID- 8247827 TI - Value judgements. PMID- 8247828 TI - Letting go. Interview by Toni Turner. PMID- 8247829 TI - In praise of intolerance. PMID- 8247830 TI - Resuscitation: revival techniques. PMID- 8247832 TI - Midwifery: the last stand? PMID- 8247831 TI - Backwards steps. PMID- 8247834 TI - Do-it-yourself development. PMID- 8247833 TI - Community care act: uncertain support. PMID- 8247835 TI - Nursing pioneers: the Peplau legacy. Interview by Phil Barker. PMID- 8247836 TI - Right to privacy for staff with HIV urged. PMID- 8247837 TI - HIV/AIDS: positive steps. PMID- 8247838 TI - Practice nursing. Progress report. PMID- 8247839 TI - Practice nursing. Leading role. PMID- 8247840 TI - Catheters: finding the best solution. PMID- 8247841 TI - Nursing Times open learning programme. M7: evaluation. Part (i): The principles of evaluation (continuing education credit). PMID- 8247842 TI - AIDS aetiology and HIV-1. PMID- 8247843 TI - Barred from safe sex. PMID- 8247844 TI - Spelling it out. PMID- 8247845 TI - A healthy performance? PMID- 8247846 TI - Sick with laughter. PMID- 8247847 TI - Family care: sweet persuasion. PMID- 8247848 TI - Client-centred care. New directions. PMID- 8247849 TI - Client-centred care. A chance for change. PMID- 8247850 TI - Strategies for motivation. PMID- 8247851 TI - Uninvited guests. PMID- 8247852 TI - Skirting the issue. Mind your back. PMID- 8247853 TI - The day after a death. PMID- 8247854 TI - Tragedy at sea. Interview by Daloni Carlisle. PMID- 8247855 TI - Nursing development units: a question of choice. PMID- 8247856 TI - Tea and sympathy. PMID- 8247857 TI - Council to take a firmer line on HIV. PMID- 8247858 TI - First-line management. Disciplined approaches. PMID- 8247859 TI - First-line management. Contact point. PMID- 8247860 TI - First-line management. Rights and responsibilities. PMID- 8247861 TI - The knowledge base for teaching and facilitating. (i) The teacher as an expert. PMID- 8247862 TI - Remanded in safe custody. PMID- 8247863 TI - Shedding tiers. PMID- 8247864 TI - An official crisis. PMID- 8247866 TI - Systems of life. The ear and hearing 1. PMID- 8247865 TI - The clean gang. PMID- 8247867 TI - Standard share. PMID- 8247868 TI - Expanding roles. Trading places. PMID- 8247869 TI - Acupressure. A point of pressure. PMID- 8247870 TI - Patient education. Turning over a new leaflet. PMID- 8247871 TI - Picking up the pieces. PMID- 8247872 TI - Transplants. Protocol for loss. PMID- 8247873 TI - Bridges, barges and babies. PMID- 8247874 TI - Community care. At risk--of what? PMID- 8247875 TI - Infection control. Beating the bug. AB - Strains of Staphylococcus aureus resistant to methicillin were first described soon after the introduction of this antibiotic. Methicillin, the first of the anti-staphylococcal penicillins, was soon replaced by cloxacillin and flucloxacillin, which have the advantage of good oral absorption, but strains of Staphylococcus aureus that are resistant to methicillin are invariably resistant to cloxacillin, flucloxacillin and the cephalosporins. A number of outbreaks of MRSA occurred in the 1960s, but the problem disappeared in the 1970s, which has been referred to as 'the decade of complacency'. In the early 1980s MRSA re emerged as a significant cause of hospital-acquired infection in many countries, including England (London area), Eire (Dublin), Australia (Perth), Saudi Arabia and the USA. Epidemic strains of MRSA are distinguished by the ability to colonise skin very effectively. They are spread rapidly between patients via the hands of medical, nursing and paramedical staff. Most district general hospitals no longer have an isolation ward; patients with infections are generally nursed in side rooms. PMID- 8247876 TI - Infection control. Lotions and potions. PMID- 8247877 TI - Infection control. Quality initiative. PMID- 8247878 TI - The knowledge base for teaching and facilitating. (ii) The teacher as a learner. PMID- 8247879 TI - Going forward. PMID- 8247880 TI - Spiritual sickness. PMID- 8247881 TI - Men and misconduct. PMID- 8247882 TI - Research methods. Informed action. PMID- 8247883 TI - Research methods. Learning from the past. PMID- 8247885 TI - Shiatsu. PMID- 8247884 TI - Unsafe as houses? PMID- 8247886 TI - Teamwork. A pivot for the practice. PMID- 8247887 TI - Emotional responses to paediatric hospitalisation. AB - This paper reports on an investigation of the emotional adjustment by children and their parents to paediatric hospital admission for planned ear, nose and throat surgery. It discusses the first phase of that adjustment and the importance of the initial clinic consultation in shaping subsequent coping. The study developed directly from clinical practice and the findings are expected to feed back into practice. The assessment of an experimental approach to preparation, which tests the efficacy of a fresh appraisal of the way in which children and their parents are prepared for this elective surgery, is studied. While there are important findings that relate to all three phases of the experience, that is, the pre-admission, in-hospital and post-hospital adjustment phases, the impact of the initial clinic consultation alone is discussed in relation to the implications for practice arising from the study. PMID- 8247888 TI - Advocacy. Balance of power. PMID- 8247890 TI - Midwives' journal. Big sister is watching you. PMID- 8247889 TI - The big chance. PMID- 8247891 TI - Midwives' journal. The joy of research. PMID- 8247892 TI - Midwives' journal. Attendance not compulsory. PMID- 8247893 TI - Maybe the last editorial on hand-over-mouth technique. PMID- 8247894 TI - HOME: learning process? PMID- 8247895 TI - HOM: more limitations. PMID- 8247896 TI - Midazolam: a review of its use for conscious sedation of children. PMID- 8247897 TI - Hereditary epidermolysis bullosa: oral manifestations and dental management. AB - Epidermolysis bullosa (EB) is a diverse group of disorders that have as a common feature blister formation with tissue separation occurring at variable depths in the skin and/or mucosa depending on the specific EB type. There may be marked oral involvement, potentially creating devastating alterations in the soft and hard tissues. Oral tissue fragility and blistering is common to all EB types. However, oral debilitation as a result of soft tissue scarring is primarily limited to the recessive dystrophic EB subtypes. Generalized enamel hypoplasia appears to be limited to junctional EB, although rampant dental caries is associated with many individuals having generalized recessive dystrophic EB. While systemic treatment remains primarily palliative, it is possible to prevent destruction and subsequent loss of the dentition through appropriate interventions and dental therapy. The majority of individuals with mild EB subtypes may receive dental treatment with only minor modifications in approach. Even the most severely affected individuals with EB can retain their dentition using general anesthesia and conventional restorative techniques. With aggressive preventive interventions and management of developing malocclusions using serial extraction, it also is possible to reduce the likelihood of rampant caries, achieve an acceptable occlusion without the need for active tooth movement or appliance therapy, and allow these individuals to benefit from maintaining a natural healthy dentition. PMID- 8247898 TI - A long-term followup on the retention rate of zinc oxide eugenol filler after primary tooth pulpectomy. AB - A retrospective study of all the patients' records (> 6000) in a pediatric dental practice was done to assess ZOE retention after a pulpectomized primary tooth was lost and the succedaneous tooth erupted. There were 65 children with 81 ZOE pulpectomies done in 30 incisors and 51 molars. Pulpectomies were done at a mean chronologic age of 52.2 months and followed for a mean time of 90.8 months from time of placement. The initial radiograph after the pulpectomized tooth was lost, showed retained ZOE filler particles in 49.4% of the cases while 27.3% had retained ZOE a mean time of 40.2 months after pulpectomy tooth loss. Short-filled pulpectomies retained significantly less ZOE than long fills (P = 0.04). With time, retained ZOE particles either resorbed completely or showed reduction of filler size in 80% of the cases. No pathology was associated with the retained ZOE particles. Retention of ZOE was not related to pulpectomy success (P = 0.11), preoperative root resorption (P = 0.76), age of the patient (P = 0.24 incisors; P = 0.87 molars), extraction/exfoliation of the pulpectomy (P = 0.75), or timing of the pulpectomy's loss (P = 0.72). PMID- 8247899 TI - An in vitro comparison of three endodontic techniques for primary incisors. PMID- 8247900 TI - Clinical and radiographic evaluation of partial pulpotomy in carious exposure of permanent molars. PMID- 8247901 TI - Evaluation of the novel anti-inflammatory agent tetrandrine as a pulpotomy medicament in a canine model. AB - Tetrandrine, a bisbenzylisoquinoline alkaloid with unique broad-spectrum anti inflammatory properties, was evaluated as a pulpotomy medicament in a canine model. Histological sections were evaluated after three days (acute inflammation) and six weeks (chronic inflammation) by two criteria: 1) intensity and degree of inflammation, and 2) extent of pulp involvement. The results of the three-day dressings revealed significant neutrophil infiltration in only 30% of teeth treated with tetrandrine, compared with 81%, 84%, and 100% of teeth treated with Ledermix, (Lederle Pharmaceuticals, Wolfrathausen, Germany), formocresol (Creighton Pharmaceuticals, Sydney, Australia) and saline (controls) respectively (P < 0.01). After six weeks, there was significant lymphocyte infiltration in only 30% of teeth treated with tetrandrine, compared with 66%, 90%, and 100% on teeth treated with Ledermix, formocresol, and saline controls respectively. (P < 0.01). In both three-day and six-week specimens in tetrandrine-treated teeth the extent of inflammation was limited to less than one-third of the coronal section of the pulp, whereas teeth treated with Ledermix or formocresol showed cellular infiltration extending to greater than two-thirds of the pulp (P < 0.01). Comparative studies with berbamine, a natural analog of tetrandrine, showed that it was less potent than tetrandrine, but significantly better than Ledermix and formocresol on both acute and chronic pulp inflammation (P < 0.05 and P < 0.01 respectively). These results suggest that tetrandrine may have value as a pulpotomy medicament. PMID- 8247902 TI - The use of behavior management techniques by dentists across practitioner type, age, and geographic region. AB - The purpose of this project was to describe the behavior management procedures employed for child dental patients based on practitioner type (general dentist vs. pediatric dentist), age, and geographic location. A survey of practice characteristics, which included questions relating to child behavior management, was mailed once to a national random sample of 3000 dentists--2000 general dentists and 1000 pediatric dentists. The pediatric dentists who responded employed a broader spectrum of management techniques than did the general dentists. Significant regional and practitioner age differences were reported in the use of behavior management techniques. The use of sedation and general anesthesia was reported more frequently in the western regions. Of the five age groups, the 40- to 49-year-old age group reported using the broadest spectrum of behavior management techniques. The survey revealed practitioner type, age, and regional differences in the child behavior management procedures employed by dentists. PMID- 8247903 TI - Shear bond strength of a resin composite to enamel treated with an APF gel. AB - The purpose of this study was to evaluate the effect of an APF gel treatment on the shear bond strength of a resin composite to enamel. A total of 45 noncarious human extracted permanent molars were used. A flat enamel surface was obtained with 600 grit silicon carbide paper and cleansed with a rubber cup and a water slurry of fine flour of pumice. The teeth were randomly distributed into three groups of 15 teeth each: Group 1: Etched for 30 sec with 37% orthophosphoric acid gel (control); Group 2: APF treatment (Topex gel--Sultan Dental Products, Englewood, NJ) for 1 min and then etched as in Group 1; Group 3: APF gel treatment for 4 min and then etched as in Group 1. After etching, rinsing, and drying, an unfilled resin (XR-Bond--Kerr Manufacturing Co., Romulus, MI) was applied thinly with a brush and cured for 20 sec. A nylon ring (internal diameter: 6.69 mm2) was placed over the area and filled with a light-cured composite resin (Herculite XR--Kerr Manufacturing Co., Romulus, MI). The teeth were thermocycled (x500), mounted in plastic cups and plaster, and sheared with a knife-edged blade in an Instron Testing Machine (Instron Engineering Corp., Canton, MA) running at a crosshead speed of 0.5 mm/min. The results in MPa were as follows: Group 1: 24.88 +/- 6.16; Group 2: 23.20 +/- 5.11; Group 3: 21.29 +/- 8.44. An ANOVA and Student-Newman-Keuls procedure revealed no statistically significant difference among the groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247904 TI - Progressive systemic sclerosis in a child: case report. AB - A case of pediatric progressive systemic sclerosis is reported and a literature review concerning medical and dental aspects of this condition is provided. Systemic features include sclerodactyly, Raynaud's phenomenon, telangiectasia, calcinosis, myositis, arthritis, tenosynovitis, renal failure, esophageal hypomotility, pulmonary fibrosis and heart failure. Oral manifestations include reduced interincisal distance, xerostomia, telangiectasia, increased periodontal ligament width, osseous resorption of the mandible, periodontal disease, and increased decayed, missing, and filled teeth (DMFT). The prognosis is difficult to predict because spontaneous remission has been documented, but death may result from extensive visceral involvement (heart, kidney, and lung). PMID- 8247905 TI - Hairy leukoplakia in a child with AIDS--a rare symptom: case report. PMID- 8247906 TI - Management of traumatic oral-facial injury in the hemophiliac patient with inhibitor: case report. AB - This report describes identification of Factor VIII inhibitor in a patient who then received immune tolerance therapy. The precipitating event was a traumatic orofacial injury that was nonresponsive to traditional factor-replacement therapies. An inhibitor complicates medical and dental management of the hemophiliac patient because it counteracts usual techniques of hemorrhage control using coagulation agents derived from Factor VIII (Monoclate--Armour, Blue Bell, PA). Successful identification and management of the inhibitor patient require communication and consultation between the physician and dentist, up-to-date knowledge regarding the hemophiliac patient's bleeding and infusion history and aggressive application of local adjunctive hemostatic therapies. PMID- 8247907 TI - An overview of the distinction among the processes of accreditation, licensure, and certification. PMID- 8247908 TI - Neglected pathogens: bacterial infections in persons with human immunodeficiency virus infection. A review of the literature (1). AB - Bacterial infections, including those that cause infection in the healthy host as well as those that are more opportunistic, occur very commonly among persons infected with the human immunodeficiency virus (HIV). Bacterial infections are a direct result of the severe humoral and cellular immune defects found in these patients. Epidemiologic factors such as intravenous drug use and stage of HIV infection may also play important roles. Pulmonary, bloodstream, gastrointestinal, central nervous system, skin and soft tissue, and catheter related infections are common, as are endocarditis, prostatitis, and others. Frequently reported pathogens are common organisms such as Staphylococcus aureus, Haemophilus influenzae, Streptococcus pneumoniae, and enteric gram-negative pathogens, as well as less typical ones such as Listeria monocytogenes and Nocardia sp. The frequency of infection is specific to organ system and pathogen, often being many times higher than in immunocompetent hosts. Prompt recognition and aggressive therapy are required to reduce morbidity and mortality due to these infections. PMID- 8247909 TI - The use of angiotensin-converting enzyme inhibitors after acute myocardial infarction. AB - During and immediately after myocardial infarction (MI), many interrelated and complex processes manifest the body's attempt to minimize damage and compensate for lost cardiac function. Although these compensatory responses may provide some short-term restoration of function, their long-term consequences actually may increase morbidity and mortality. Several agents have established roles in the treatment of these patients, whereas others, including the angiotensin-converting enzyme (ACE) inhibitors, have yet to be investigated thoroughly. Results of two trials investigating the role of ACE inhibition after MI seem to provide sufficient data to warrant the use of these drugs in certain patient populations. These results are promising, but further investigation is necessary to answer key questions arising from these trials. PMID- 8247910 TI - Immunosuppressive properties of calcium channel blockers. AB - Calcium channel blockers are extensively used for their beneficial effects on the cardiovascular system. Solid organ transplant recipients commonly have cardiovascular disease and are often treated with these agents. Research demonstrates that calcium antagonists may have beneficial effects in this population that are independent of their effects on the cardiovascular system. Indeed, both in vitro and in vivo studies suggested that they may possess immunosuppressive properties. Their actions at the cellular level in both the afferent and efferent arms of the immune system indicate that alone, as well as in combination with cyclosporine, these agents possess immunosuppressive properties that may potentially benefit the transplant population. PMID- 8247911 TI - Administration of insulin by continuous ambulatory peritoneal dialysis. AB - Many patients with diabetic nephropathy undergoing continuous ambulatory peritoneal dialysis (CAPD) use their peritoneal access to administer insulin. Compared with the subcutaneous route, intraperitoneal (IP) insulin may display more consistent absorption, produce more physiologic insulin concentrations, and be more convenient to administer. However, there are no well-controlled trials that have demonstrated a clinically significant difference in glycemic control between IP and subcutaneous administration. For patients who choose to begin IP insulin at the time CAPD is initiated, the starting dose is 2-3 times the previous subcutaneous dose. For patients previously stabilized on CAPD, the conversion factor may be less. Doses are divided equally between bags. Some authors recommend adding more insulin to bags with a higher concentration of dextrose. In addition, the dose should be decreased when added to a bag used for an overnight dwell. Exchanges performed during the day may be timed to start 30 minutes before a meal. Unless clinical trials demonstrate a difference in efficacy between subcutaneous and peritoneal insulin administration, the route will remain a matter of patient preference. PMID- 8247912 TI - A follow-up safety study of ciprofloxacin users. AB - We followed 37,233 outpatients for 45 days after receiving a prescription for ciprofloxacin to identify any newly diagnosed, important illnesses that might have been caused by the drug. For 29 users the role of ciprofloxacin in the etiology of the illness could not be confidently ruled out (7.79/10,000 persons; 95% CI 5.42-11.18). In only seven was a causal relation to ciprofloxacin considered likely: three skin reactions and one case each of thrombocytopenia, "headache, nausea, and shakes," hallucinations, and palpitations. No fatal illnesses occurred, and all patients recovered after discontinuing the drug. In addition, few cases of photosensitivity were associated with the agent. We conclude that important adverse reactions attributable to ciprofloxacin are uncommon. PMID- 8247913 TI - Cefepime pharmacokinetics in cystic fibrosis. AB - STUDY OBJECTIVE: To determine the disposition of cefepime in patients with cystic fibrosis compared with healthy controls. DESIGN: Open-label, single-dose study. SETTING: Laboratoire de Pharmacocinetique Clinique, Universite Laval, Quebec, Canada. PATIENTS AND SUBJECTS: Twelve patients with the confirmed diagnosis of cystic fibrosis (CF) and 12 healthy volunteers. One subject with CF withdrew for personal reasons; the data of another patient were excluded from the evaluation of renal values due to incomplete urine collection. INTERVENTIONS: A single 2000 mg dose of cefepime was administered as a 30-minute intravenous infusion. Healthy subjects did not use any other drugs throughout the study. Those with CF refrained from taking prophylactic antibiotics prior to and during the study, but continued to use pancreatic enzymes, multivitamins, and beta-agonist and/or steroid inhalers. One patient continued insulin treatment. MEASUREMENTS AND MAIN RESULTS: Cefepime's maximum concentration was approximately 150 micrograms/ml at the end of the infusion, half-life 2-2.5 hours, and urinary recovery 80% in both groups. No statistically significant difference was seen in any of the pharmacokinetic values between the groups, except for the mean residence time (2.03 +/- 0.26 vs 2.39 +/- 0.37 hrs; p < 0.02). Total clearance was 19% higher in patients with CF than in healthy volunteers (119.7 +/- 20.1 vs 103.5 +/- 19.8 ml/min), perhaps due to higher renal (95.1 +/- 12.4 vs 85.1 +/- 12.0 ml/min) and/or nonrenal (25.4 +/- 13.1 vs 18.4 +/- 12.0 ml/min) clearances in subjects with CF. CONCLUSIONS: The disposition of cefepime is not significantly affected by CF, and dosage adjustment appears not to be necessary in these patients. PMID- 8247914 TI - The disposition and cerebrospinal fluid penetration of morphine and its two major glucuronidated metabolites in adults undergoing lumbar myelogram. AB - STUDY OBJECTIVE: To determine the pharmacokinetic disposition of morphine and its two major glucuronidated metabolites, morphine-6-glucuronide (M6G) and morphine-3 glucuronide (M3G), in serum and their penetration into cerebrospinal fluid (CSF). DESIGN: A single-dose, open-label pharmacokinetic study. SETTING: The Memphis Neurosciences Center at Methodist Hospital. PATIENTS: Twenty patients undergoing a diagnostic lumbar myelogram. INTERVENTIONS: All patients received morphine sulfate 10 mg intramuscularly approximately 1.5 hours before the procedure. MEASUREMENTS AND MAIN RESULTS: Three blood samples were drawn after the dose and a CSF sample was drawn immediately after lumbar puncture. The mean +/- standard deviation for the half-life of morphine was 2.8 +/- 1.4 hours. The apparent half lives of M6G and M3G were 5.7 +/- 3.1 and 6.3 +/- 2.2 hours, respectively, and were inversely related to the estimated creatinine clearance (r = -0.61, p < 0.007 and r = -0.69, p < 0.002, respectively). The mean concentrations of morphine, M6G, and M3G in the CSF (collection time 1.5 +/- 0.32 hrs, n = 19) were 3.1 +/- 3.7, 12.5 +/- 17.6, and 19.6 +/- 16.1 nmol/L, respectively. CONCLUSIONS: Elderly patients and patients with renal dysfunction receiving morphine may experience prolonged analgesic and adverse effects secondary to a decrease in the clearance of M6G. PMID- 8247915 TI - Safety of different dosages of intravenous adenosine used in conjunction with diagnostic myocardial imaging techniques. AB - STUDY OBJECTIVE: To determine the safety of three different dosage regimens of intravenous adenosine. DESIGN: Open-label, observational safety evaluation. SETTING: University hospital-based department of nuclear medicine. PATIENTS: Cohort of 854 patients referred for myocardial perfusion imaging to evaluate their coronary artery disease and who were judged unable to perform physical exercise. INTERVENTIONS: Subjects underwent myocardial perfusion imaging in conjunction with one of three intravenous dosage regimens: 1 = fixed dosage 140 micrograms/kg/minute for 6 minutes; 2 = dosage titration to a maximum of 140 micrograms/kg/minute; and 3 = dosage titration to a maximum of 200 micrograms/kg/minute. In regimens 2 and 3, maximum tolerated dosages were continued for a minimum of 3 minutes prior to radioisotope injection. MEASUREMENTS AND MAIN RESULTS: Adenosine-induced hemodynamic, electrocardiographic, and biochemical changes were measured. Adverse effects of the different adenosine dosages were compared. Noncardiac side effects such as flushing, dyspnea, neck tightness, and lightheadedness occurred at a significantly higher rate during regimens 2 and 3 than regimen 1. Chest pain and first- and second-degree atrioventricular block were also more frequent during regimens 2 and 3. However, the frequency of other side effects such as complete heart block, hypotension, and ST segment depression did not differ among the regimens. High-dose adenosine was associated with a significant increase in serum uric acid, a significant decrease in blood glucose, and a significant increase in serum triglyceride levels. Mean changes in hemodynamics and electrocardiographic intervals were also not different among the groups except for a greater increase in PR interval in regimens 2 and 3 than regimen 1. Discontinuation of adenosine was infrequent (< 1%) and did not differ among the regimens. CONCLUSIONS: Adenosine-assisted myocardial perfusion imaging procedures are relatively safe for evaluating coronary artery disease. Despite 82% of patients reporting at least one side effect, only 10 (< 1%) had to discontinue adenosine. No patient suffered any residual sequelae from the adverse effects. The fixed-dose regimen is associated with fewer subjective side effects and is better tolerated than titration regimens. Appropriate safety precautions should nonetheless be taken during adenosine infusions. PMID- 8247916 TI - Racial differences in the pharmacokinetics of methylprednisolone in black and white renal transplant recipients. AB - STUDY OBJECTIVE: To examine the comparative pharmacokinetics of long-term methylprednisolone therapy in black and white renal transplant recipients. DESIGN: Comprehensive pharmacokinetic evaluations of patients who participated in our glucocorticoid-monitoring program. SETTING: University-based renal transplantation clinic. PATIENTS: Six white renal transplant recipients with stable renal function, sex- and (approximate) age-matched with six preselected black patients. INTERVENTIONS: The daily oral methylprednisolone dose for each patient was administered intravenously, and serial plasma samples were obtained over 24 hours. MEASUREMENTS AND MAIN RESULTS: Methylprednisolone was analyzed by high-performance liquid chromatography. The drug's pharmacokinetics in black and white patients, respectively, were as follows: mean clearance 234 +/- 124 and 472 +/- 180 ml/hr/kg (p < 0.05); volume of distribution 0.3-2.0 and 0.8-2.0 L/kg; and elimination rate constant 0.13-0.41 and 0.27-0.42 hour-1 (p < 0.06). No statistical difference was noted in the last two parameters. The mean half-life of 3.4 +/- 1.4 hours in black patients compared with 2.1 +/- 0.3 hours in white patients approached statistical significance (p < 0.08). CONCLUSIONS: These preliminary observations suggest that the disposition of methylprednisolone differs between black and white renal transplant recipients. The current method of prescribing glucocorticoids employs a fixed-dose regimen that does not take these possible interracial differences into consideration. Incorporating the differences may allow for more individualized dosing and more efficacious use of the agent in this patient population. PMID- 8247917 TI - A randomized, controlled trial comparing diltiazem, hydrochlorothiazide, and their combination in the therapy of essential hypertension. AB - STUDY OBJECTIVES: To compare the efficacy of combination therapy with sustained release diltiazem and hydrochlorothiazide (DTZ SR-HCTZ) with that of monotherapy with DTZ SR, HCTZ, or placebo in the treatment of essential hypertension; and to determine whether the addition of a diuretic to diltiazem at apparent optimum doses of each agent significantly enhances their antihypertensive effects. DESIGN: Multicenter, randomized, double-blind, placebo-controlled, parallel-group trial with a 6-week treatment phase. SETTING: Private and university-based clinics. PATIENTS AND PARTICIPANTS: Subjects of either sex, ranging in age from 18-70 years, with a diagnosis of stable essential hypertension made from two consecutive weekly mean supine diastolic blood pressure (DBP) readings of 95 mm Hg or above to 110 mm Hg or less that varied 7 mm Hg or less after 4-6 weeks in the baseline phase. Of the patients enrolled, 298 met the inclusion criteria. INTERVENTIONS: Combination therapy with DTZ SR-HCTZ 120 mg-12.5 mg, or monotherapy with DTZ SR 120 mg or HCTZ 12.5 mg, or placebo was administered twice daily. MEASUREMENTS AND MAIN RESULTS: Combination therapy with DTZ SR-HCTZ lowered both supine DBP and SBP significantly (p < 0.005) more than either single agent. The combination also lowered DBP and SBP significantly more than either monotherapy. During a 12-hour in-clinic monitoring period spanning a dosing interval, both the combination and DTZ SR therapies maintained efficacy, whereas the antihypertensive effects of HCTZ dissipated after 8 hours. Treatment-related adverse events for the combination and HCTZ were similar but slightly greater than those for DTZ SR and placebo. CONCLUSIONS: The addition of a diuretic to sustained-release diltiazem produced an enhanced antihypertensive effect compared with monotherapy with either individual agent. PMID- 8247918 TI - Prevention of auditory sequelae in pediatric bacterial meningitis: a meta analysis. AB - Meta-analysis methods were used to compare the effect of antibiotic regimens and corticosteroids on the development of auditory sequelae after pediatric bacterial meningitis. After a literature search of two major data bases, 11 studies met the criteria and were included in the analysis. Summary odds ratios, with 95% confidence intervals, were calculated that quantified the relative risk of developing auditory sequelae after specific therapies. No significant differences among antibiotics were identified in terms of reducing the risk of meningitis related hearing impairment. However, corticosteroids significantly reduced the frequency of bilateral, moderate, or greater hearing loss. Patients receiving placebo versus dexamethasone were much more likely to develop auditory dysfunction (odds ratio 3.77; 95% CI 1.77-8.10). The results of this study add quantitative evidence supporting the use of dexamethasone as adjunctive therapy in pediatric bacterial meningitis. PMID- 8247919 TI - Faculty have few incentives for becoming board certified in pharmacotherapy. AB - A telephone survey of all colleges of pharmacy in the United States (including Puerto Rico) and Canada was conducted to assess the incentives offered for pharmacy practice faculty to become board-certified pharmacotherapy specialists (BCPS). Board certification is not a requirement for employment at any college of pharmacy; however, it plays a part in promotion, tenure, and merit salary increases at several schools. Fewer than half of the schools provide at least partial reimbursement of examination-related expenses. Sixteen have initiated BCPS study groups. We perceive that faculty must have more incentives to take the examination. It may be difficult to change institutional policies regarding merit increases or promotion and tenure, but colleges of pharmacy can do more to decrease the cost burden and to promote study groups within the department. PMID- 8247920 TI - Predictors of trough concentrations of oral ciprofloxacin. AB - Patients enrolled in a fixed-dose clinical trial of oral ciprofloxacin had trough concentrations measured to document absorption and monitor compliance. The objective was to determine whether any demographic characteristics might be important predictors of the concentrations. Stepwise multivariate linear regression revealed no correlation between ciprofloxacin trough concentrations and serum creatinine, estimated creatinine clearance (Clcr), weight, height, body surface area, or gender. However, age exhibited a direct linear relationship with trough concentrations (Y in microgram/ml), Y = 0.020.age--0.541 (p < 0.003). We conclude that for patients with Clcr 30 ml/minute or above, age is a more important predictor of ciprofloxacin trough concentration than renal function. Dosage adjustment should not be arbitrary but should be guided by minimum inhibitory concentration, clinical response, and side effects. PMID- 8247921 TI - Possible interaction between warfarin and fluconazole. AB - Fluconazole is a triazole antifungal agent reported to have a low affinity for human cytochrome P-450, and thus would not be expected to interact with drugs metabolized through the cytochrome P-450 system, including phenytoin, cyclosporine, and warfarin. A potential interaction between warfarin and fluconazole occurred in a 39-year-old man with chronic renal insufficiency. He was receiving anticoagulant therapy for a lower extremity thrombus and oral fluconazole 50 mg/day for a fungal urinary tract infection. After attaining consistent international normalized ratio (INR) values between 2.0 and 2.7 with warfarin, the INR increased to 5.2 four days after fluconazole was started, despite decreasing the dosage of warfarin. There were no changes in the patient's other medications, and the INR decreased to 1.5 on discontinuation of fluconazole. The possible mechanism of an interaction may be dose-related inhibition of warfarin metabolism, and may be more pronounced in patients with decreased renal clearance of fluconazole. PMID- 8247922 TI - Neuroleptic malignant syndrome associated with clozapine monotherapy. AB - Neuroleptic malignant syndrome is thought to be a result of dopamine receptor blockade in the striatum. Clozapine has only weak affinity for dopamine type 1 and 2 receptors, and therefore it was thought this drug would not precipitate the syndrome. However, six cases of the syndrome have been reported in patients receiving clozapine monotherapy. A review of the pathoetiology of symptoms occurring in the syndrome is included. PMID- 8247923 TI - Fetal exposure to lisinopril: neonatal manifestations and management. AB - The use of angiotensin-converting enzyme (ACE) inhibitors in pregnancy has been associated with neonatal morbidity and mortality. The mechanism of renal dysfunction likely is related to fetal hypotension and prolonged decreased glomerular filtration. Six of 14 previously published cases of neonatal renal failure after maternal ACE inhibitor therapy resulted in death. Eight infants survived after peritoneal dialysis, some with residual renal impairment. Serum lisinopril levels and ACE activity in our patient indicate that during the anuric state the drug has an extremely prolonged half-life, and that it is removed by peritoneal dialysis. In view of this prolonged half-life and the drug's continued suppression of ACE activity and renal function, we recommend institution of early dialysis in infants with renal failure after maternal therapy with lisinopril. PMID- 8247924 TI - The one who dies with the most letters behind his name wins. PMID- 8247925 TI - Author's affiliations. PMID- 8247926 TI - Ethics in clinical pharmacy practice and research: an introduction. PMID- 8247927 TI - Ethical issues related to clinical pharmacy research. American College of Clinical Pharmacy. PMID- 8247928 TI - Pharmacists and the pharmaceutical industry: guidelines for ethical interactions. American College of Clinical Pharmacy. PMID- 8247929 TI - Women as research subjects. American College of Clinical Pharmacy. PMID- 8247930 TI - HIV-infected children in the pediatric emergency department. AB - Forty-three nonhemophiliac, confirmed HIV-positive children followed by the Children's Hospital AIDS Program made 184 visits to the children's Emergency Department (ED) during 1988 and 1989. The mean age was 30 +/- 28 months with a median of 25 months, a mode of 10 months, and a range from two days to 19 years. CD4 counts from within six months of the visit were available in 87% and were low enough to require Pneumocystis carinii pneumonia prophylaxis under current guidelines in 52%. Chief complaints included fever in 50%, respiratory symptoms in 21%, and gastrointestinal symptoms in 8%. The ED discharge diagnosis included fever/possible sepsis in 25%, pneumonia in 17%, otitis media in 9%, and upper respiratory tract infection or viral syndrome in 9%. Overall, an acute infection was identified at 62% of visits; of these, 33% were judged to be serious in nature. A total of 92 blood cultures were drawn, of which eight were positive with the following organisms: Streptococcus pneumoniae (3), Streptococcus faecalis (2), Escherichia coli (1), Torulopsis glabrata (1), and Staphylococcus non-aureus (1, a probable contaminant). Overall, 53% of patient encounters resulted in hospitalization. Patients with a white blood cell count more than 15,000/mm3 were more likely to be hospitalized (87 vs 62%, P < 0.01), though the white blood cell count was not helpful in identifying patients with bacteremia or serious infections. The mean temperature of patients admitted was higher than in those discharged (38.7 vs 37.9 degrees C, P < 0.01). In 1989, an estimated six per 1000 visits to our facility were by HIV-infected children. PMID- 8247931 TI - Critical care pediatrician-led aeromedical transports: physician interventions and predictiveness of outcome. AB - This article reviews the one-year experience (March 28, 1987 to March 27, 1988) of the pediatric transport service of the University of Wisconsin Hospital and Clinics (UWHC). The UWHC pediatric transport team consisted of a critical care flight nurse and a pediatric critical care attending physician or fellow. The aims of the study were to: 1) determine the types and number of interventions performed by the physicians to gauge the need for physician presence on transport; and 2) determine which variables (severity of illness scores, age, gender, distance from hospital) recorded at the time of the referral telephone call best predicted outcome of the patient. There were 109 children transported by helicopter during the one-year study period. Thirty percent of the patients (43% of trauma patients and 22% of medical patients) had no interventions at all, 18% of medical patients and 10% of trauma patients were intubated, and 9% of medical patients and no trauma patients had central venous catheters inserted. Multivariate modeling determined that among medical patients, outcome could be accurately predicted only 38% of the time if telephone PRISM (Pediatric Risk of Mortality) scores were determined. Among trauma patients, if gender, age, distance from UWHC, and telephone PRISM scores were known, outcome could be predicted 74% of the time. Unless studies show the benefit of pediatrician accompanied transport, transports could probably be done without critical care pediatricians. Severity of illness scoring at this time is probably not sufficiently accurate to warrant its use for deciding the appropriateness of transport of pediatric patients. PMID- 8247932 TI - The educational value of pediatric emergency transport: by design or by default? AB - Pediatric emergency transport teams provide an important and challenging clinical experience in many residency training programs. However, the educational value of this experience has not been carefully examined. A 32-item questionnaire was developed to assess curricular components of the three phases of the transport experience in residency training: phase 1 included pretransport training and qualifications and curricular planning, phase 2 addressed intratransport phone backup for the team analogous to online medical command, and phase 3 included posttransport feedback and performance evaluation. The study also addressed the chief residents' perception of the roles of the resident and transport nurse working on the team. The survey was mailed to chief residents at 76 pediatric residency programs in the United States. Data were received from 75 programs for a return rate of 98%. Fifty-four programs that used residents for transport team staffing were analyzed using descriptive statistics. Of the respondents, 39% report a specific training process prior to transport. Twenty-two percent of programs provide residents with written materials such as a manual or course syllabus. Eighty-two percent have a senior physician always identified for phone backup. Fifty-two percent of programs have no specific method for team evaluation and 31% rely only on informal verbal feedback. Thirty percent routinely provide feedback within one week of the transport. Sixty-three percent of chief residents view the experienced transport nurse as an equal member of the transport team. The study found that the educational structure of transports is largely unplanned and that basics such as preservice training and timely performance feedback are not usually addressed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247933 TI - Tuberculous meningitis in children. PMID- 8247934 TI - Cerebral abscess formation following ocular trauma: a hazard associated with common wooden toys. AB - Penetrating orbital trauma is an uncommon pediatric emergency with potentially devastating consequences. We report a five-year-old child who fell on a wooden toy and sustained an orbital roof fracture. He subsequently developed an intracranial abscess. Current information pertaining to penetrating orbitocranial trauma will be reviewed, stressing the importance of early recognition of the intracranial injury in these cases. Both parents and toy manufacturers must remain aware of the potential for injury involving these common toys. PMID- 8247935 TI - Soaring suppurative sea shells from the sea shore: Pseudomonas aeruginosa and Klebsiella pneumoniae septic arthritis after a marine sea shell injury. AB - Septic arthritis is the most important diagnosis to consider in patients presenting with acute monarticular arthritis. We present the case of an eight year-old girl who developed a Pseudomonas aeruginosa and Klebsiella pneumoniae septic arthritis of her knee following an injury with a marine sea shell. After treatment with antibiotics and arthroscopic irrigation, she had good functional recovery. We could find no previously reported cases of Pseudomonas aeruginosa or Klebsiella pneumoniae septic arthritis resulting from an injury in a marine environment. The pathogenesis and treatment of septic arthritis and infection following marine injuries are discussed. PMID- 8247936 TI - Small doses, big problems: a selected review of highly toxic common medications. AB - Many commonly used medications have serious toxicity in children when ingested in small doses. The toxicologic characteristics of methyl salicylate, camphor, topical imidazolines, benzocaine, and diphenoxylate-atropine are striking examples. All of these medications except Lomotil are over-the-counter and therefore, are often perceived as minimally harmful when ingested. For all of these substances, however, doses as little as 1/4 teaspoon or 1/2 tablet can have serious or fatal consequences. Thus, referral to an emergency department is prudent for ingestions involving these products. Options for initial gastrointestinal (GI) decontamination are variable, depending on the estimated amount and time of the ingestion. Induction of emesis is contraindicated for significant camphor, topical imidazoline, and Lomotil ingestions. Activated charcoal should be administered in all cases. Finally, the emergency physician must recognize the potential seriousness of these ingestions, as well as their clinical presentations to provide expeditious evaluation and treatment. PMID- 8247937 TI - Treatment of paraphimosis using the "puncture" technique. AB - The puncture technique is a simple method that aids in the manual reduction of the paraphimotic foreskin. We present a case that describes how we are currently using the technique in our pediatric patients. PMID- 8247938 TI - Telephone advice in the pediatric emergency department. PMID- 8247939 TI - Nausea, vomiting, and diarrhea in an older child. PMID- 8247940 TI - Case 04-1993: an adolescent female with abdominal pain and iron deficiency anemia. PMID- 8247941 TI - Elbow injury. PMID- 8247942 TI - Torsade de pointes. PMID- 8247943 TI - [Possible role of serotonin in hemostatic the mechanism of action of desmopressin (DDAVP) in patients with uremia]. AB - Haemorrhagic diathesis is a serious complication of uraemia. Desmopressin is known to shorten prolonged bleeding time in uraemia but mechanism of the haemostatic action of this drug remains still unknown. The aim of the work was to study the effect of desmopressin on some haemostatic parameters in relation to plasma and platelet serotonin. Desmopressin was administered i.v. to 33 haemodialysed patients (age range 27-66 years) in a dose of 0.4 microgram/kg b.w., 90 minutes after desmopressin infusion bleeding time became significantly shorter (p < 0.001) and correlated with the shortening of the euglobulin clot lysis time (r = -0.43, p < 0.05). Tissue plasminogen activator activity increased (p < 0.01) and its inhibitor (PAI) activity decreased (p < 0.001) after desmopressin infusion. A correlation between the fall in platelet serotonin content and changes in tissue plasminogen activator and PAI activities was found (r = 0.55, p < 0.01 respectively). A rise in plasma serotonin concentration was observed. In vitro desmopressin inhibited 14C serotonin uptake in a dose dependent manner. After 2 hours of platelet incubation with desmopressin in a concentration of 4 ng/ml 16% of 14C serotonin was released. A possibility of serotoninergic mechanism in the haemostatic action of desmopressin is suggested. PMID- 8247944 TI - [Evaluation of humoral immune response in patients with asymptomatic and symptomatic HIV infection. Analysis of titers of anti-Hbs, antibodies against cytomegalovirus, herpes simplex virus type I, rubella virus and toxoplasma gondii]. AB - 40 asymptomatic HIV carriers and 45 AIDS patients were tested for anti-HBs (Hepatitis B Virus surface antigen), anti-RV (Rubella virus), anti-Toxo (Toxoplasma gondii), anti-CMV (Cytomegalovirus) and anti-HSV-1 (Herpes simplex virus type 1) antibody titers and compared with 83 persons characterized by risk behaviours but seronegative for HIV. The prevalence of these antibodies was very high and similar in all three groups studied, however, patients with AIDS had generally lower antibody titers when compared with asymptomatic carriers. The only exception being anti-HSV-1 which was present in high titre even in gravely ill patients. It seems that subjects with clinically overt HIV infection develop a serious disturbance in the humoral immune response with depressed specific antibody synthesis. PMID- 8247945 TI - [Cytoenzymatic and ultramicroscopic investigations of blood in coke oven workers]. AB - White blood cell count, cytoenzymology (acid and alkaline phosphatase, beta glucuronidase and myeloperoxidase of granulocytes) and ultrastructure of granulocytes and NBT reduction test were performed in peripheral blood obtained from cokery plant workers. All the subjects were divided into groups according to degree of exposure to BaP. Occupational exposure to many factors during coke production, especially to high concentration of BaP cause perceptible changes of NBT reduction test in the group more exposed workers. A statistically significant of the totally activity of the acid phosphatase and beta-glucuronidase of granulocytes were found in this risk group. The changes in granulocyte function correlated with ultrastructural changes. The coking plant environment represent a strong stimulator of the neutrophil metabolism. PMID- 8247946 TI - [Results of alternating treatment of multiple myeloma--cytostatics VBMCP and recombinant interferon alfa-2B]. AB - In 19 patients with recently diagnosed multiple myeloma 3-week cycles of vincristine, BCNU, melphalan, cyclophosphamide and prednisone alternating with interferon were administered over 6-12 months. Results were compared with a control group of 33 myeloma patients treated exclusively with VBMCP cytostatics. In interferon treated patients objective response was more frequent (76%) and median survival time longer (above 35 months). PMID- 8247947 TI - [Interferon alfa in treatment of chronic hepatitis B in children with leukemias and lymphomas]. AB - The course of the hepatitis B virus infection (HBV) in the majority of children with cancer who have undergone intensive chemotherapy is characterized by scanty symptoms and often leads to chronic hepatitis and prolonged carrier state. Twenty three children with chronic hepatitis B virus (HBV) were treated with interferon alfa after the completion of chemotherapy of leukemias and lymphomas. In the majority of cases a disproportion between symptomless clinical course of the hepatitis B virus and the rate of its active replication was observed. Presence of HBsAg and high activity of polymerase DNA HBV (pDNA) were found in all children. In the course of a 6-month treatment with interferon in a dose of 3 mil IU/m2 of the body surface a gradual decrease of pDNA activity was observed. A statistically significant difference of mean values of pDNA before and right after the treatment was determined (p < 0.05). Tests performed 3 months after the completion of the therapy showed a repeated increase of pDNA activity. Boundary values of pDNA were observed in 5 children. Complete elimination of HBs and HBe antigens has not been achieved in any of the children. While analyzing the obtained results it has to be taken into consideration that there were numerous unfavourable prognostic factors determined in the studied group of children such as: coexisting malignant disease, undergone intensive chemotherapy, long period which has passed from the moment of HBV infection until the administration of the treatment with interferon, high activity of pDNA, scanty symptoms of inflammation process. Coexisting HCV infection in 12 children, could also have a undesirable effect upon the treatment results.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8247948 TI - [Effect of parathormone (PTH)on blood pressure]. PMID- 8247949 TI - [A new presynaptic histamine H3 receptor]. PMID- 8247950 TI - [Primary biliary cirrhosis--diagnosis and therapeutic aspects]. PMID- 8247951 TI - [Influence of chronic ketanserin therapy on blood pressure and certain humoral and metabolic factors in patients with mild to moderate primary essential hypertension]. AB - Ketanserin, a serotonin-2-receptor antagonist, was administered to 16 patients with mild to moderate essential hypertension in randomized single-blind placebo controlled study. After 2 weeks of placebo administration ketanserin 60 mg daily was given for 3 weeks. In 7 patients the normalization of blood pressure was obtained and they were given the same daily dose of ketanserin for another 3 weeks. In the remaining 9 patients the dose of ketanserin was increased to 120 mg daily for the same period; in one additional case blood pressure decreased to normal. In the whole group the decrease of diastolic pressure was statistically significant comparing to placebo period. Ketanserin therapy did not influence serum free serotonin concentration, but the hypotensive effect was greater in patients with lower serotonin blood levels. Serum aldosterone concentration decreased in the initial period of treatment; the decrease was more pronounced in patients who responded to therapy. In this subgroup of patients diastolic blood pressure correlated positively with plasma renin activity. Ketanserin lowered serum adrenaline concentration, particularly in patients with good response to therapy. There was also decrease of total cholesterol concentration without any significant changes in HDL--cholesterol fraction. The authors conclude that oral chronic ketanserin treatment is an effective therapy in essential hypertension and that its effectiveness is greater in patients with lower serum serotonin levels. The hypotensive effect of the drug may be connected with lower sympathetic activity and with changes in renin-angiotensin-aldosterone system. Ketanserin is well tolerated and exerts beneficial effect on lipid metabolism. PMID- 8247952 TI - Congenital bilateral juvenile granulosa cell tumor of the ovary in leprechaunism: a case report. AB - We report on a case of leprechaunism. In addition to the typical clinical and biochemical features, a bilateral juvenile granulosa cell tumor of the ovaries and cytomegalovirus hepatitis were found. The granulosa cell tumor may result from the mitogenic effect of insulin at high concentration, which acts via a mechanism mediated by insulin-like growth factor I receptors. PMID- 8247953 TI - Adenoid cystic carcinoma of the lacrimal gland in a six-year-old girl. AB - A 6-year-old white female was found to have an adenoid cystic carcinoma originating in a lacrimal gland. Eighteen months following diagnosis, the tumor recurred. Conservative surgery has been the sole mode of therapy. To date, after four operations and quadrimenstral imaging surveillance, there is no sign of disease progression. Our purpose is to record the unusual occurrence of an adenoid cystic carcinoma of the lacrimal gland in a child. An interim report, 32 months after diagnosis, is presented. PMID- 8247954 TI - An unusual lateral neck cyst with the combined features of a bronchogenic, thyroglossal, and branchial cleft origin. AB - Soft tissue cystic masses arising in the neck in children are commonly of embryologic origin and often surgically excised. Depending on location and histology, they can be classified as thyroglossal duct, branchial cleft, or rarely bronchogenic in origin. We present an unusual case of an infant with a lateral neck mass that communicates with the midtrachea. The histologic appearance is that of a combination of the features seen in cysts of branchial cleft and thyroglossal duct origin. The possible embryologic development of such a lesion is discussed. PMID- 8247955 TI - Lethal manifestations of neurofibromatosis type 1 in childhood. AB - Neurofibromatosis type 1 (NF-1), also known as peripheral neurofibromatosis or von Recklinghausen's disease, is a common inherited neurocutaneous syndrome. Complications of the disease are numerous and may be fatal, but because the frequency of potentially lethal complications increases with age, deaths from the disease are often thought to occur only in adults. Examination of autopsy records at the Adelaide Children's Hospital for the period 1952 to 1991 revealed four cases of children with NF-1. These cases illustrate a number of interesting pathologic features including a proliferative glial infiltrate involving the subarachnoid space, two primary gliomas within the same patient, gliomatosis cerebri, and sudden death due to massive intratumoral hemorrhage. PMID- 8247956 TI - Neonatal and infantile pulmonary hemorrhage: an autopsy study with clinical correlation. AB - We studied the clinicopathologic features of pulmonary hemorrhage in autopsied infants less than 1 year of age for a 12-month period. There were 70 liveborns (LB) and 24 stillborns (SB). The percentage of LB with pulmonary hemorrhage (PH), pulmonary interstitial emphysema (PIE), hyaline membrane disease (HMD), acute bronchopneumonia (ABP), congenital malformations (CM), and surgery (SUR) were analyzed according to weeks of estimated gestational age (EGA) and as an entire group. Overall, 74% of LB and 24% of SB had histologic evidence of PH. A semiquantitative evaluation of the extent of PH among the LB infants disclosed that hemorrhage involved less than one-third of the observed lung tissue in 42%, one-third to two-thirds of the lung was hemorrhagic in 15%, and the remaining 42% had more than two-thirds hemorrhagic lung parenchyma. A total of 31 clinical and pathologic factors were evaluated for their possible association or relationship to PH. Statistical analysis revealed that hyaline membranes and hemorrhage in other extrapulmonary sites were the only significant associations with PH. PH was more frequent in premature infants born between 24 and 39 weeks EGA and was always associated with multiple other conditions. Although the autopsy finding of hemorrhage in the lungs is relatively frequent in the population we studied, it appears that PH as a primary phenomenon in infants is extremely rare, if it occurs at all. PMID- 8247957 TI - Performance of reticulocyte counts in stored blood specimens in a pediatric population utilizing new methylene blue. AB - Although reticulocyte counts can be reliably performed for up to 48 h after storage in EDTA, it is unclear whether this is applicable to the pediatric age group. In order to evaluate this, manual reticulocyte counts were performed on 20 specimens from pediatric patients stored at 4 degrees C for up to 24 h post collection. Samples were evaluated at 1-3, 6, 12, 18, and 24 h after storage in EDTA vacutainer tubes at 4 degrees C. The age of the subjects ranged from 1 day to 9 years with a median age of 3 years. Patients' reticulocyte counts ranged from 0 to 27% (5.89 +/- 7.21). No clinically significant changes were evident in the reticulocyte count over 24 h after specimen collection. The mean of the 20 specimens at 1-3 h was 5.50 and at 24 h was 5.40 (P > .05). The standard deviation of the mean values ranged from 7.03 to 7.26 (P > .05). The results indicate that reticulocyte counts may be performed on previously drawn blood held at 4 degrees C for up to 24 h post collection in a pediatric population without significant difference from baseline values. PMID- 8247958 TI - In utero development of pulmonary artery calcification in monochorionic twins: a report of three cases and discussion of the possible etiology. AB - Pulmonary artery (PA) calcification is unusual in children, reportedly occurring only in conjunction with severe underlying valvular disease. Three newborns were found by chest X-ray study to have thin calcified rings of the PA. Two were pump twins in twin reversed arterial perfusion (TRAP) sequences, each with an acardiac cotwin, and the third was the recipient in a twin transfusion syndrome (TTS). Both twin pairs with TRAP sequence were premature, each pump twin presenting with cardiac decompensation. One remained well at 5 months of age, the other died at 5 1/2 months of age. The third, the premature recipient of twins with TTS, died at 24 h of age, and the donor cotwin died at 2 h of age. The two hearts were structurally normal at autopsy. Subtle intimal wrinkling was seen above the pulmonary valve leaflets, and the media was yellow and granular on cut section. Hyperplasia of the intima and media with disruption and calcification of the elastic fibers of the media was seen microscopically. These three cases of PA calcification occurring in utero were not related to structural valvular abnormalities but were presumably due to increased cardiac output in utero as each occurred in the volume-overloaded twin of the twin pair. Comparison of the weight differences between these three twin pairs with PA calcification suggests a relationship between the presence of PA calcification and the severity of the cardiac volume overload. In utero cardiac damage may contribute to the high morbidity and mortality rate seen in infants with TRAP sequence and TTS. PMID- 8247959 TI - Successful prenatal therapy of thoracic lesions. AB - Improvements in prenatal imaging techniques now permit in utero treatment of potentially serious lesions. We describe three cases in which decompression of significant space-occupying thoracic lesions resulted in an excellent clinical outcome. The pregnancies were all carried to term after second or early third trimester intervention and resulted in infants with minimal respiratory distress. Cytologic and chemical analysis of the thoracic fluid aspirated in utero provided diagnostic and physiologic information. PMID- 8247960 TI - Angiomatous malformation of placental chorionic stem vessels and pseudo-partial molar placentas: report of five cases. AB - Five placentas referred to the Michigan Placental Tissue Registry between 1982 and 1992 exhibited an unusual malformation of chorionic stem vessels. Multiple intestine-like tangles arising from umbilical vessels rested on and entered the chorionic plate. This anomaly appears similar to the "unusual vascular anomaly of the placenta" reported by Lee et al. (Am J Clin Pathol 1991; 95:48-51). In addition, amnionic-type bands were often adherent to the vascular complex. In three cases, clusters of macrocystic villi interspersed by normal placental parenchyma resulted in a partial molar appearance. Two of the infants were stillborn; three were liveborn. Congenital anomalies were not recognized in other organs. Flow cytometric analysis of placental tissue from four specimens disclosed a diploid chromosomal pattern. This rare anomaly complex appears to be a malformation involving the extraembryonic mesoderm rather than a cytogenetic error. PMID- 8247961 TI - Dystrophin analysis in the diagnosis of childhood muscular dystrophy: an immunohistochemical study of 75 cases. AB - Seventy-five consecutive pediatric muscular dystrophy (MD) cases were analyzed by dystrophin immunohistochemistry (75/75), Western blot analysis (26/75), DNA analysis (30/75), and immune electron microscopy (8/75). The patients included 64 males and 11 females and the clinical diagnoses were Duchenne MD (DMD) (41), Becker MD (BMD) (8), intermediate/outlier MD (4), female DMD (3), limb girdle or Becker (1), congenital MD (CMD) (10), Fukuyama CMD (1), facioscapulohumeral MD (FSH) (3), limb girdle MD (2), and other uncharacterized dystrophies (2). Dystrophin analysis was performed on all cases using the N- and C-terminal antidystrophin antibodies. Dystrophin analysis helped to exclude an Xp21 dystrophy in four patients. Except for two patients who showed normal staining with the N-terminal and abnormal staining with the C-terminal antibody, all DMD cases showed absent staining except for the immunoreactive revertant fibers, which were generally under 5%. A variety of staining patterns was seen in BMD, ranging from normal to abnormal (variable intensity of staining, partially stained/unstained fibers). Abnormalities were observed with the C-terminal antibody in one case of CMD and Fukuyama CMD, and normal staining was present in the other dystrophies. Immune electron microscopy confirmed absence of staining in DMD and normal membrane staining in other dystrophies. Our study underscores the importance of using antibodies with specificities to different regions of the dystrophin molecule for accurate diagnosis. As abnormal staining may be encountered in non-Xp21 dystrophies such as CMD, dystrophin staining should not be used in isolation to make a diagnosis of a dystrophinopathy. PMID- 8247962 TI - Congenital malformations: a review of 672 autopsies in Ibadan, Nigeria. AB - The postmortem findings in 672 Nigerian children with lethal congenital malformations are reviewed. Eighty-six percent of the patients were less than 1 year of age and the male-to-female sex ratio was 1.3:1. The alimentary tract was the most common anatomic site involved (30%), intestinal and biliary atresia being the two lesions most frequently identified in this location. Twenty-six percent of the children had cardiovascular malformations, of which ventricular septal defect, patent ductus arteriosus, and tetralogy of Fallot were most important. There was an accentuated male predominance among cases with urogenital malformations due to the high frequency of posterior urethral valve. Hydrocephalus, spina bifida, and anencephaly were the most common lesions observed in the craniospinal axis. One-fifth of the children had complex malformations of multiple organ systems, 20% of which were associated with chromosomal anomalies, rubella, and other malformation syndromes. PMID- 8247963 TI - Sudden death in infants under one year of age. AB - Clinical and postmortem findings on 126 consecutive cases of sudden infant deaths occurring in Metropolitan Toronto over a 42-month period were reviewed and classified. Of these cases, 101 were diagnosed as sudden infant death syndrome (SIDS), the other 25 as due to other causes. The SIDS cases were classified into three groups: A-1 (39%), infants born at term who died between 2 weeks and 6 months of age and in whom clinical and autopsy findings were negative; A-2 (4%), same as A-1, except that the infants were born prematurely (< 36 weeks of gestation); and B (57%), infants with miscellaneous minor clinical and pathological findings not directly related to or responsible for death. The most common findings in group B were clinical and/or pathological signs of minor respiratory infection. These findings were prevalent in older SIDS infants (> 10 weeks at death), with up to 80% concordance between clinical and pathological indices of respiratory infection. Group B also included cases with minor congenital abnormalities, drug use by mother, and other miscellaneous findings. This report defines the clinicopathological features of SIDS infants and describes the spectrum of findings associated with the syndrome. It is suggested that by applying uniform criteria for definition and classification of SIDS a more reliable data base can be obtained, facilitating research into the causes and pathogenesis of SIDS. PMID- 8247964 TI - A congenital variant of glycogenosis type IV. AB - Three related patients are described with glycogenosis type IV with an unusual clinical presentation resulting in perinatal death. Stored material showed birefringent Maltese crosses and was present in skeletal muscles, heart, central nervous system, and liver. Muscular dysfunction resulted in a fetal hypokinesia sequence with arthrogryposis and lung hypoplasia. A subdivision of glycogenosis type IV in four subtypes is proposed, based on age of onset. Measurement of the enzyme activities in different tissues does not permit, at the moment, a distinction between the subtypes. PMID- 8247965 TI - In situ fixation of the neonatal brain and spinal cord. AB - A delay in the autopsy can result in significant tissue autolysis, especially in the central nervous system. We have developed a rapid technique of in situ fixation that preserves central nervous system tissues until the formal autopsy can be performed. Through the lateral margin of the anterior fontanelle, Zamboni's solution is injected percutaneously into the lateral ventricles and allowed to exit via an intrathecal spinal needle. The choice of fixative allows a wide array of postmortem studies to be done. PMID- 8247966 TI - PREParation for PRCP. PMID- 8247967 TI - Immunization for hepatitis B infection: interest, information, and initiative? PMID- 8247968 TI - Assessment and treatment of childhood obesity. PMID- 8247969 TI - Sinusitis. PMID- 8247970 TI - Gastrointestinal endoscopy in infants and children. PMID- 8247971 TI - Index of suspicion. Case 1. Diagnosis: cocaine abuse. PMID- 8247972 TI - Index of suspicion. Case 2. Diagnosis: myastheniagravis. PMID- 8247973 TI - Index of suspicion. Case 3. Diagnosis: congestive heart failure due to dilated cardiomyopathy. PMID- 8247974 TI - Disorders of the spinal cord. PMID- 8247975 TI - When bigger isn't better. Interview by Sarah L. Gall. PMID- 8247976 TI - Who will advocate for patients?. Interview by Amber Stenger. PMID- 8247977 TI - Change is coming to doctor-patient relationships. Interview by Lisa Schnirring. PMID- 8247978 TI - Rx for healthcare: let patients pay their share. Interview by Amber Stenger. PMID- 8247979 TI - Will personal freedom survive reform?. Interview by Robert J. Roos. PMID- 8247980 TI - The reform-plan maze. A quick look at three ways to go. PMID- 8247981 TI - Decoded, the President's plan is bad medicine for Americans. PMID- 8247982 TI - Patients worry about rationing and long lines. Interview by Robert J. Roos. PMID- 8247983 TI - Medicine for the masses: what suffers?. Interview by Lisa Schnirring. PMID- 8247984 TI - Managed care can be quality care. Interview by Corrine Charais. PMID- 8247985 TI - Is old-fashioned patient care obsolete?. Interview by James R. Wappes. PMID- 8247986 TI - Patients can bid farewell to 'Cadillac' healthcare. Interview by Flora Taylor. PMID- 8247987 TI - Rural medicine: rocky soil for managed care?. Interview by Robert J. Roos. PMID- 8247988 TI - Will reform limit access for rural patients?. Interview by Michael D. Toft. PMID- 8247989 TI - Headache in elderly patients. How to recognize and manage benign types. AB - Headaches in elderly persons are usually benign. Nevertheless, the possibility of underlying organic disease increases with age. To facilitate diagnosis and determine suitable treatment, primary care physicians should become familiar with diagnostic criteria for common causes of headache in elderly persons. Thorough physical examination and history taking can help rule out many underlying diseases. Benign headaches can usually be managed successfully with available non pharmacologic and/or pharmacologic interventions. When organic disease (eg, temporal arteritis, cerebrovascular disease, tumor) is suspected, referral to an appropriate specialist may be indicated. PMID- 8247990 TI - Chronic pelvic pain. Differentiating anatomic from functional causes. AB - Patients with chronic pelvic pain are challenging, from both diagnostic and therapeutic viewpoints. At the pain clinic at the Medical College of Georgia School of Medicine, Augusta, we have discovered over the past 3 years that psychiatric disease is probably underrecognized in these patients and that physicians are often reluctant to discuss intimate details of a patient's sex life even though they may provide important information. In addition, sleep disorders are very prevalent. Use of vaginal-probe ultrasound may decrease the need for diagnostic laparoscopy, especially in patients without pelvic disease. In most cases, ovarian cysts in ovulating patients do not represent an abnormality. Laparoscopy and major pelvic surgery should be limited to patients with a high probability of anatomic abnormalities or persistent symptoms. PMID- 8247991 TI - Kaposi's sarcoma mimicking cellulitis. AB - Although Kaposi's sarcoma is not the most common cause of death in AIDS patients, it is often one of the initial opportunistic illnesses associated with human immunodeficiency virus infection. Extensive plaque formation and edema in the lower extremities may take on the appearance of cellulitis, and in dark-skinned persons, the lesions of the neoplasm may not be noticeable. Treatment is palliative; therapy for local effect is appropriate unless lesions are extensive or systemic involvement is present. PMID- 8247992 TI - Drug dosing in patients with renal insufficiency. A simplified approach. AB - Because renal impairment interferes with the disposition of drugs, the dosage of many medications needs adjustment to prevent excess morbidity and prolonged hospital stays. Physiologic considerations in patients with renal insufficiency include serum albumin levels, creatinine clearance, volume status, cardiac function, and body weight. Dosing errors in the hospital can be minimized by using (1) a computerized system of reporting serum creatinine levels and (2) a pharmacokinetic monitoring service to assure optimal benefit from a drug without its toxic effects. Drug interactions are common and may alter the metabolism of other agents; therefore, medication lists should be updated and reviewed regularly. PMID- 8247993 TI - Rheumatoid arthritis. Importance of early diagnosis in long-term outcome. AB - Rheumatoid arthritis is a common disorder that does not have the favorable outcome it was once thought to have. A detailed clinical history, thorough physical examination, and judicious use of serologic tests and radiologic studies should allow accurate diagnosis, even at an early stage. Traditional therapy can successfully improve short-term functional status but may not alter long-term outcome. Early, aggressive therapy may improve prognosis, but this approach must be further studied and, if attempted, should be done in consultation with a rheumatologist. PMID- 8247994 TI - Say no to 1,342 pages of more government control. PMID- 8247995 TI - Snow jobs. PMID- 8247996 TI - What will happen to me? PMID- 8247997 TI - What will happen to me? PMID- 8247998 TI - What will happen to me? PMID- 8247999 TI - How big a dose of levothyroxine? PMID- 8248000 TI - Aggressive treatment of acute myocardial infarction. Management options for various settings. AB - Multiple lifesaving options are currently available for treatment of acute myocardial infarction as a medical emergency. Serial electrocardiography and continuous ST-segment monitoring, urgent echocardiography, rapid enzyme analysis, and cardiac catheterization may all assist in the accurate and early diagnosis of acute myocardial infarction. Both intravenous thrombolytic therapy and direct infarct percutaneous transluminal coronary angioplasty are of benefit in early treatment. The choice of therapy depends on the individual patient and the hospital capabilities. Adjunctive pharmacologic therapies can be easily administered in the community hospital setting and should be considered for every patient with suspected acute myocardial infarction. The risk of serious morbidity and hospital death in these patients has not been eliminated, and a more aggressive approach to diagnosis and treatment is sorely needed. PMID- 8248001 TI - Chronic bronchitis. Key points in evaluation. AB - Thorough history taking is essential in evaluation of chronic bronchitis. Patients often reveal key symptoms that help define the disorder and provide information about contributing factors, such as cigarette smoking, that can be eliminated. Useful baseline data can be collected through pulmonary function studies, electrocardiogram, chest radiographs, complete blood cell count, and measurement of electrolyte levels. PMID- 8248003 TI - American Association of Pharmaceutical Scientists, AAPS 8th Annual Meeting and Exposition. Lake Buena Vista, Florida, November 14-18, 1993. Abstracts. PMID- 8248002 TI - Chronic bronchitis. Choosing the optimal treatment. AB - Among the various therapies for chronic bronchitis none is more important, both for relieving symptoms and for preserving pulmonary function, than cessation of cigarette smoking. Unfortunately, even when patients are motivated and programs are aggressive, results are unspectacular. Chronic bronchitis often responds favorably to bronchodilating agents. For initial therapy, ipratropium bromide (Atrovent) is the agent of choice because of its efficacy and safety. The role of antiinflammatory drugs is not yet clear, although inhaled steroids are beneficial in some patients. Most patients improve with smoking cessation and/or judicious pharmacologic intervention. PMID- 8248004 TI - Glucagon produced by recombinant DNA technology: repeated dose toxicity studies, intravenous administration to CD rats and beagle dogs for four weeks. AB - The toxicity of glucagon produced by recombinant DNA technology (Glucagon (ge)) was studied by daily intravenous administration to rats and dogs for 4 weeks. Pancreatic glucagon of bovine or porcine origin (Glucagon Novo) was used as a reference control in the dogs. Glucagon (ge) has the same sequence of the 29 amino acids as pancreatic glucagon of humans, cows, pigs, rats and dogs. The dosages were 0 (control), 0.2, 1.0 and 5.0 mg Glucagon (ge)/kg/day in the rats, and 0 (control), 1.0 and 5.0 mg Glucagon (ge) and 5.0 mg Glucagon (Novo)/kg/day in the dogs. The studies complied with current EEC, US and Japanese guidelines for 4 week toxicity studies of drugs. All dose levels were well tolerated. The plasma glucose and cardiovascular responses to dosing were monitored in the dogs and found to be in agreement with well-known effects of pancreatic glucagon. The most consistent finding in both species was an increase in liver weight. This change was without concomitant pathological deviations in the other parameters examined. There were no differences in the reaction of dogs following treatment with Glucagon (ge) or Glucagon (Novo). A dose of 1 mg Glucagon (ge)/kg/day was regarded as a clear no-toxic-effect-level in both species. PMID- 8248005 TI - Effects of amitraz on motor function. AB - The effects of amitraz, a formamidine derivative, on motor function were studied in rats. Behavioural and neurochemical studies were performed. Results show that amitraz was able: 1) to decrease locomotion and rearing frequencies of rats and to increase their immobility time in an open-field; 2) to displace to the left the control dose-response curve constructed to apomorphine-induced stereotyped behavior; 3) to potentiate both pentobarbital sleeping time and amphetamine effects on open-field behaviour of rats and 4) to increase not only the whole brain levels of noradrenaline but also the striatal levels of dopamine. In relation to control animals, the pesticide also induced 5) a decrease in the striatal levels of homovanillic acid, 6) pesticide effects on open-field behaviour of rats were not antagonized by yohimbine administration, and 7) metoclopramide administration on amitraz pretreated rats antagonized not only amphetamine effects on open-field behaviour, but also apomorphine-induced stereotypy. These results are discussed in the light of the actions of amitraz on biogenic amines. It is suggested that the pesticide effects on motor function are consequences of the inhibitory effects on MAO activity, most probably through the increases produced on catecholamine levels within the central nervous system. PMID- 8248006 TI - Influence of pH on the uptake and pharmacodynamics of quinidine in the isolated perfused rat heart. AB - Using the single-pass isolated perfused rat heart preparation we examined the effect of perfusate pH (pH 7.05, 7.46, 7.71, 7.92) on quinidine output concentration (C(out)) and delta QT. Eight hearts were perfused at 2.5 ml/min. with quinidine (20 microM) for 35 min. followed by a 35-40 min. washout period with drug-free perfusate. This procedure was repeated four times in each preparation with the pH sequence varied and the same pH used in the first and last phases. Increasing pH slowed the rate of equilibration of C(out), the equilibration rate constant (k) decreasing from 0.273 min.-1 at pH 7.05 to 0.095 min.-1 at pH 7.92. A modified Kety-Renkin-Crone equation was fitted to the C(out) versus time data for each pH. The estimated volume of distribution (V) increased significantly with pH from 11.5 +/- 1.1 to 32.5 +/- 2.9 ml/g, but the permeability surface product did not change with pH (mean 17.7 ml/min./g). There was a linear relationship between V and calculated un-ionised quinidine C(out), with an intercept of 5.70 ml/g corresponding to the V of ionised drug. This indicates that ionised and un-ionised drug readily enter the heart and that the slower equilibration with pH is due to the increased V which results from increased partitioning of un-ionised quinidine into myocardial tissue. Perfusion pH did not directly affect baseline QT interval, but the rate of attainment of maximum delta QT decreased with increasing perfusate pH. Plots of delta QT versus calculated coronary output quinidine concentration did not change with pH, showing that this drug effect was due to both ionised and un-ionised moieties. This study shows that myocardial permeability and pharmacodynamic effect (delta QT) of quinidine are not influenced by perfusion pH over the range 7.0 to 7.9, although rate of equilibration of both C(out) and effect vary with pH. PMID- 8248007 TI - Pharmacokinetics and pharmacodynamics of pentazocine in children. AB - Pharmacokinetics and ventilatory effects of a single intravenous dose of 0.5 mg/kg of pentazocine were studied in ten children aged 4 to 8 years after ophthalmic surgery. Elimination half-life (mean +/- S.D.) was 3.0 +/- 1.5 hr and clearance 21.8 +/- 5.9 ml/min./kg. The values for Vc, Vss and V beta were 0.73 +/ 0.21, 4.0 +/- 1.2 and 5.3 +/- 2.1 l/kg, respectively. The pharmacokinetic parameters were similar to those of adults. After administration of pentazocine decrease in ventilatory rate and oxygen saturation and increase in end-tidal carbon dioxide were relatively fast and steep. Oxygen saturation of four patients decreased below 90% and in one patient the decrease did not recover instantly and additional oxygen was given for 2 min. No patient needed assisted ventilation. Only clinically insignificant changes in heart rate and mean arterial pressure were observed. The duration of analgesia was 164 +/- 59 min. No serious side effects appeared. PMID- 8248008 TI - A simple HPLC method for the determination of cellular glutathione. PMID- 8248009 TI - Vitamin A, a useful biochemical modulator capable of preventing intestinal damage during methotrexate treatment. AB - The effects of vitamin A on methotrexate treatment of mice inoculated with L1210 leukaemia cells or sarcoma 180 cells were studied. The small intestine of tumour transplanted mice was severely damaged after methotrexate treatment. Coadministration of vitamin A with methotrexate protected the small intestine from methotrexate-induced damage. The protective effect of vitamin A was histologically and biochemically demonstrated. Further, coadministration of vitamin A did not inhibit in vivo antitumour activity of methotrexate. Thus, biochemical modulation used to prevent methotrexate-induced malabsorption by vitamin A coadministration will be of great use in methotrexate cancer chemotherapy. PMID- 8248010 TI - Interaction of myeloperoxidase and elastase enzyme activity with the antihypertensive agents hydralazine and dihydralazine. AB - Autoantibodies directed against myeloperoxidase and elastase have been found in patients developing hydralazine-induced lupus and hydralazine-induced isolated glomerulonephritis. The aim of this study was to investigate influence of hydralazine and dihydralazine upon myeloperoxidase and elastase enzyme activity. Using a 4-aminoantipyrin in vitro system, dihydralazine was 2.5 times as potent in inhibiting myeloperoxidase activity as compared to hydralazine. The corresponding Ki-values were 4 microns M for dihydralazine and 25 microM for hydralazine. When using 2.2'-azino-bis-3-ethyl-benzothiazoline-6-sulphonic acid system inhibition was found at lower concentrations. Furthermore, the difference between the compounds was not so pronounced as seen for 4-aminoantipyrin. The Ki values for hydralazine and dihydralazine were 1.2 and 1.4 microM respectively. Complete inhibition was seen for both compounds at concentrations above 7.5 microM. Hydralazine binds to elastase, but neither hydralazine nor dihydralazine inhibited elastase enzyme activity. PMID- 8248011 TI - Detomidine binding to guinea pig liver imidazoline receptors (I-receptors) shows marked positive cooperativity. AB - The binding of [3H]-idazoxan to guinea pig liver membranes was measured in the presence of 3 microM rauwolscine, which prevented the binding [3H]-idazoxan to alpha 2-adrenergic receptors. Under these conditions the radioligand bound to saturable imidazoline receptors (I-receptors) with a Kd of 18 nM and a Bmax of 665 fmol mg-1 protein. Six drugs which were used to compete for [3H]-idazoxan in the liver caused competition curves of widely varying steepness. Fitting the competition curves to the standard four parameter logistic function showed that the Hill coefficients (nH) varied from 2.02 (detomidine) to 0.43 (UK-14,304), The nH's obtained in liver for the six compounds correlated strongly (r = 0.99; P < 0.001) with the corresponding nH's obtained in a previous study on the guinea pig kidney where the drugs were also tested in competition with [3H]-idazoxan (Wikberg et al. 1992). Good correlation was also found for the Log(Ki) values of drugs determined in the two tissues (r = 0.96; P < 0.005). Whereas the standard logistic function accurately described the competition curves of the 5 drugs tested in the liver for which the competition curve Hill coefficients varied between 0.43 to 1.41 (UK-14,304, rilmenidine, histamine and d- and l medetomidine), it did not accurately fit the detomidine competition curves. Instead the detomidine competition curves could be more accurately described by a model composed of the sum of two independent logistic functions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248012 TI - Investigation of acute cardiovascular and respiratory toxicity of HLo 7 dimethanesulfonate and HI 6 dichloride in anaesthetized guinea-pigs. AB - The bis-pyridinium dioxime HLo 7 is considered to possess promising therapeutic properties in the treatment of organophosphate poisoning. Acute circulatory and respiratory effects of HLo 7 and HI 6 were therefore compared in anaesthetized guinea-pigs. Female Pirbright white guinea-pigs were anaesthetized with urethane and the carotid artery, jugular vein and trachea were cannulated. Saline or atropine, 10 mg/kg, or HLo 7 or HI 6 (30 or 100 mumol/kg, each) or atropine plus oxime were injected intravenously after base line measurements. Respiratory and circulatory parameters were recorded for 60 min., then blood was drawn for AChE measurement. Injection of HLo 7 or HI 6 alone resulted in a temporary, dose dependent hypotension, an almost unchanged heart rate and a slight respiratory stimulation. A more severe hypotension appeared after the administration of atropine plus HLo 7 or HI 6. In these groups heart rate and respiration were markedly stimulated. Measurement of AChE activity in blood samples revealed no impairment by HLo 7 or HI 6 with or without atropine. These results suggest that HLo 7 has only transient effects on the cardiorespiratory system after intravenous administration and its safety regarding acute circulatory and respiratory toxicity is comparable to HI 6. PMID- 8248013 TI - Dynamic effects and pharmacokinetics of lemakalim in the isolated guinea-pig heart. AB - Myocardial effects of lemakalim were studied in the isolated retrogradely perfused and spontaneously beating guinea-pig heart. Pharmacokinetic analysis of drug accumulation and disposition showed two-compartment characteristics with phasic half-times of about 0.27 and 2.1 min., respectively. Myocardial clearance was 6.8 ml min.-1, and myocardial drug accumulation was 5-fold. Dynamic parameters were studied during increasing lemakalim concentrations from 1.4 nM up to 10 microM. Dynamic steady-states developed within 3-4 min. Increase of coronary flowrate showed an Emax of 185% with an EC50 of 60 nM. Lemakalim produced biphasic inhibitory actions on contraction velocity and amplitude. Emax and EC50 for the two phases were: 48% and 47% and 100 nM and 65 nM, respectively, for the first phase and 100% in both cases and 1.7 microM and 1.6 microM for the second phase. Heart frequency increased to a maximum of 15% above baseline at 0.3 microM. Oxygen consumption increased progressively at concentrations above 7 nM and reached a maximum of 175% at 0.3 microM. The frequency-corrected QT-interval was biphasically shortened to a maximal extent of 30% at the highest concentration. Myocardial efficiency expressed as the ratio of amplitude times frequency to oxygen consumption exhibited a progressive decline to about 20% of control values. PQ- and QRS-intervals showed only minor or no changes, respectively. No arrythmogenic effects were observed. The study demonstrated a rapid and very moderate accumulation of lemakalim in the guinea-pig heart accompanied by a marked increase in coronary flowrate and a progressive negative inotropic effect followed by reduced myocardial efficiency. PMID- 8248014 TI - Mechanistic aspects of the control of tyrosinase activity. AB - There is now much evidence suggesting that there are multiple control points in the process of melanin production. The most fundamental process of melanogenesis is centered on the oxidative activity of the enzyme tyrosinase. Tyrosinase is a highly unusual enzyme in that it apparently catalyses two processes, i.e., the oxidation of tyrosine and the dehydrogenation of dihydroxyphenylalanine (Dopa), at the same active site. The reactions involved account for the unusual kinetics of tyrosine oxidation and suggest biochemical mechanisms whereby the activity of the enzyme and the process of melanogenesis may be modified. It is proposed that the oxidative engine of melanogenesis resides in an oxidation/reduction cycle involving Dopa and dopaquinone and that this can be modified by processes that result in the removal of dopaquinone or Dopa from the reaction system. PMID- 8248015 TI - From gene to protein: determination of melanin synthesis. AB - Melanin production in mammals is regulated at a variety of levels (tissue, cellular, and subcellular), and many gene loci are involved in the determination of color patterns directed by the melanocyte. Many of the genes involved in these complex processes have now been cloned, and even the simplest mutation can lead to dramatic changes in the phenotype of the individual. Many, if not all, of the pigment related genes have pleiotropic effects on the development and differentiation of the organism, and perhaps because of this, the melanocyte is evolving as an important model for the study of gene regulation and action at the functional level. In view of the importance of pigmentation as a photoprotective barrier and as a cosmetic factor affecting appearance and social acceptance, the importance of these studies seems destined to increase significantly in the future. PMID- 8248016 TI - Does melanin turnover occur in the eyes of adult vertebrates? AB - This paper is a review of what is known about the turnover of melanin in iris, choroid, and retinal pigment epithelium (RPE) of the adult vertebrate eye. Differences in size and structure of choroideal and retinal pigment epithelial melanin granules are shown by electron micrographs. The classical stages of melanin synthesis, including the premelanosome, are shown in the RPE of adult hamsters that had been exposed to intense light. Degradation or synthesis of melanin also seem to occur in the melanocytes of the choroid in these animals. It is postulated that all three pigmented eye tissues (iris, RPE, and choroid) of adult vertebrates form melanin granules in vivo. However, nothing is known about the amount of this turnover. PMID- 8248017 TI - Pteridines as reflecting pigments and components of reflecting organelles in vertebrates. AB - This paper reviews evidence for the presence of pteridines in iridophores, leucophores, and xanthophores in a wide variety of vertebrate chromatophores, and argues that the chemical and functional distinction between pterinosomes and reflecting platelets is not as clear-cut as previously believed. Observations indicate that: (1) Pteridines may, either alone or in conjunction with purines, form pigment granules that reflect light, (2) these pigment granules are highly variable ranging from fibrous pterinosomes to typical reflecting platelets and may be colored, reflect white light, or be iridescent, and (3) many "leucophores" probably contain typical pterinosomes and presumed associated colorless pteridines and are therefore more closely related to erythrophores and xanthophores than to iridophores with which they are usually classified. We propose that the classification of pigment cells should be modified to reflect these facts. PMID- 8248018 TI - An ultrastructural study of melanocytes and melanosomes in the skin and hair bulbs of rufous albinos. AB - We have examined hair bulb and skin melanocytes of rufous albinos from Southern Africa to further characterize this form of albinism. In the skin melanocytes we find both eumelanosomes and pheomelanosomes at various stages of melanization and, in addition, there appeared to be many aberrant incompletely melanized melanosomes. On average, rufous melanosomes are 30% smaller than normal black skin melanosomes. In the keratinocytes, the melanosomes are packaged into distinct aggregations, whereas in normal black skin, they occur singly. We suggest that the reddish skin color of these albinos is a consequence of an increase in the pheomelanin synthesis resulting in a raised pheomelanin/eumelanin ratio and that the aggregation of melanosomes results in a skin color slightly lighter than normal. In hair bulb melanocytes, only eumelanosomes were seen and these were mostly incompletely melanized. These findings correlate with our visual observations that the hair color of Southern African albinos is very pale (light brown or ginger). Based on our observations, we speculate on the possible cause of rufous albinism. PMID- 8248019 TI - Light (Blt), a mutation that causes melanocyte death, affects stria vascularis function in the mouse inner ear. AB - The Light mutation (Blt) is a dominant allele of the b-locus on mouse chromosome 4 which causes progressive dilution of coat colour. Melanocytes within the hair follicles of mutant mice develop normally but later degenerate, due to the accumulation of a toxic product, so that the hair becomes lighter with age. Previous studies on W-locus spotting mutants, from which melanocytes are absent, have shown that melanocytes in the stria vascularis of the inner ear are essential for the development and/or maintenance of the endocochlear potential (EP) which is normally around 100 mV. In this study, physiological recordings from the ears of Light mutants were correlated with strial ultrastructure. EPs recorded from all b/b controls and young homozygous and heterozygous mutants (20 22 days old) were normal (77 to 113 mV), but were reduced (19 to 59 mV) in about 30% of ears from older mutants (Blt/Blt and Blt/b). Strial function therefore appears to develop normally but later degenerates in some mutants. This suggests that strial melanocytes are affected by the Light allele and that the continued presence of melanocytes is necessary for strial function. There was no obvious association between the recorded EP value and the ultrastructural appearance of the stria. No structural abnormalities of the stria were noted in control or mutant mice aged 20 days to 4 months including those which had a reduced EP. Strial atrophy was common in old controls and mutants (1-2 years), and appeared to be an age-related process rather than an effect of the Light mutation. Similarly, pigment build-up was common in all strial cells of old mice. However, the accumulations of lipofuscin-like pigment were much larger and more abundant in aged brown non-agouti mice than those observed in old agouti mice, which suggests that this age-related process also has a genetic component. PMID- 8248020 TI - Histological, biochemical, and ultrastructural studies on hyperpigmented human skin xenografts. AB - The mechanisms for hyperpigmentation observed in human cutaneous xenografts placed on athymic nude mice was investigated. Histologic, biochemical, histochemical, and ultrastructural examinations were performed on human skin prior to grafting and at various times ranging from 2 weeks to 30 weeks post grafting (PG). Hyperpigmentation was macroscopically visible on the graft as early as 4-6 weeks. The number of Dopa-positive melanocytes per unit area was increased at 2 weeks PG and remained elevated until 20 weeks PG. The surface area of the melanocytes, a measure of the activity of the cells, also increased significantly and remained above the pre-grafting size throughout the study. Western blot analysis using tyrosinase specific antibody (alpha Ty-SP) revealed the presence of tyrosinase exclusively in the grafted skin from 2 weeks to 12 weeks PG tested. Histological and ultrastructural observations revealed the presence of numerous dendritic melanocytes, indeterminant clear cells suggestive of Langerhans cells, and dermal melanophages. The results of this study suggest that the observed hyperpigmentation in grafted tissue is caused by an increase in the number of Dopa-positive melanocytes and probably from enhanced melanin production. Extracts of proteins from the xenografts exhibited prominent differences in low and high molecular proteins between pre- and post-grafted skin. Among them, the exclusive appearance of a protein doublet with apparent mw approximately 14 kDa was found in grafted skin, and subsequent studies indicate it has potent effects on melanocyte function. PMID- 8248021 TI - ["Unworthy of life"?-"A non-person"?]. PMID- 8248022 TI - [The "pediatric department" in Luneburg: murder of handicapped children between 1941 and 1945]. AB - At the end of the year 1941 a department for severely handicapped children, called "Specialized children's department" was established in the state mental hospital at Luneburg. Until 1945 695 children were admitted there, 418 of them did not survive. The majority of those children were killed--mostly by means of high dosages of barbiturate. In this article the formation of the department, the origins of the approach, the explanations of the persons involved and the experiences of the affected parents are described. PMID- 8248023 TI - ["Never useful even for the most primitive accomplishments". Murder of handicapped and sick children 1939 to 1945 in the Ueckermunde district hospital]. AB - During the Nazi era "euthanasia" of the disabled and sick was introduced with the founding of the "Reich's Commission for the Scientific Assessment of Hereditary and Genetically Determined Serious Suffering" August 1939 on 18th. The task of this commission was to organize the killing of physically and mentally disabled children in so-called "Specialized Children's Departments". In the existing literature on the killing of sick children and adolescents, reports of the "official child-euthanasia", organized by the Reich's Commission, predominate. However, independently from the Reich's Commission, many children and adolescents were murdered in the programme of adult "euthanasia". This paper will focus on the latter neglected group of victims, taking the example of the psychiatric sanatorium "Ueckermunde". A large number of participants shared responsibility, during the time of National Socialism, for the murder of disabled and sick children. Various medical specialties such as paediatrics, genetics, gynaecology, administration, general practice, psychiatry and child and adolescent psychiatry were involved. PMID- 8248024 TI - [Evaluation of an intensive therapy program for treatment of severe behavioral disorders in mentally handicapped patients with autistic or psychotic behavior]. AB - The development of severely disturbed and socially accepted behavior in mentally handicapped persons with autistic or psychotic symptoms is documented before, during and after an intensive therapy program conducted in a residential institution for mentally handicapped persons. Seven single case studies were made as long term observation with a duration between 18 and 33 weeks, mostly with a multiple baseline design. One or two follow ups with at least four weeks length were conducted in six out of seven cases up to four years after the end of the intensive therapy. The main results show (1) the decrease of disturbed behavior and the increase of socially accepted behavior during the therapy program, and (2) the significant reduction of the disturbed behavior patterns taking place during the baseline phase before the beginning of the therapy in all cases but one. The results are discussed under the aspects of a possible explanation for the findings and their impact on the discussion about psychotherapy research. PMID- 8248025 TI - [Role and responsibilities of psychosocial treatment personnel in the pediatric clinic: (I) Foundations and problem aspects of psycho-somatic cooperation]. AB - Nowadays, the core problems of psychosocial care of pediatric inpatients are those imposed by the chronically ill child and his family. In Germany, these new and complex problems are supposed to be solved by integrating psychosocial specialists and developing a comprehensive, interdisciplinary approach. The first part of this series discusses its problems and pitfalls. To find out their role in the complex network, the new specialists misleadingly may define their task as exclusively devoted to casework. The medical staff may perceive this as a kind of neglect of its town psychosocial competencies. As a consequence, they will become suspicious of the new colleagues, disregarding their potentially, too. Instead of a true cooperation, this may lead to a kind of fragmentation of the professional network. PMID- 8248026 TI - Partial purification of a pea seed DNA-binding protein that specifically recognizes 5-methylcytosine. AB - Previously, a DNA-binding protein (DBPm) was identified in plant nuclei that may mediate the effects of DNA methylation on chromatin structure and transcription. In the present report, DBPm was partially purified from germinated pea (Pisum sativum) seed nuclear extracts by DEAE-cellulose, phenylsepharose, heparin sepharose chromatography, and preparative mobility shift on polyacrylamide gels. The purified activity showed a band at approximately 50 kD by sodium dodecyl sulfate-polyacrylamide gel electrophoresis as well as by Sephadex G100 chromatography, suggesting that DBPm is present as a monomer. PMID- 8248027 TI - Isolation of human polymorphonuclear leukocyte elastase by chromatography on immobilized benzamidine. AB - Recent evidence suggests that polymorphonuclear leukocyte (PMN) elastase causes tissue injury in a variety of diseases. Current methods of purification of elastase involve several steps which result in a low yield. We report a simple purification method. PMN (10(9) in 4 ml of 0.05 M Tris, pH 7.8, containing 0.2% Triton X-100 were disrupted and homogenized by freezing and thawing followed by sonication. After centrifugation at 100,000 g for 20 min, enzyme was extracted from the pellet with 2.5 ml of 0.05 M Tris/1M NaCl (pH 7.8). The centrifugation extraction cycle was repeated 3 times. Elastase from 10(8) PMN was then purified using a 1 ml Protease Inhibitor Affinity-Filter prepared by binding benzamidine to silica. Enzyme activity was determined by cleavage of the synthetic substrate N-Suc-(Ala)3-pNa. SDS-PAGE demonstrated 2 polypeptides, molecular masses of 29 and 27 kD with amino acid composition and partial N-terminal sequence (Ile-Val Gly-Gly-Arg-Arg-Ala-Arg-Pro-His-Ala-Trp-Pro-) identical with those previously reported for elastase. We obtained 50 micrograms elastase (34-fold purification) with specific activity of 52 U/mg/min from 10(8) PMN. This represents a much greater recovery (23% yield) than is achieved by other methods. This method is simple, highly reproducible, and can be performed within a 2-day period. PMID- 8248028 TI - The histidyl-tRNA synthetase from Streptococcus equisimilis: overexpression in Escherichia coli, purification, and characterization. AB - We describe the high-level expression of the Streptococcus equisimilis histidyl tRNA synthetase gene (hisS) in Escherichia coli and the purification and characterization of the gene product. Due to a lack of an efficient E. coli ribosome binding sequence in the hisS gene, the coding region was fused in-frame to the expression vector pT7-7, thereby creating a fusion gene construct (pT7 7recIII), which is under the control of a strong bacteriophage T7 promoter. Another construct (pT-7recII) was used for low level expression of the native histidyl-tRNA synthetase (HisRS). The plasmids were electroporated into E. coli HB101, which already contained pGP1-2. After temperature induction, the fusion HisRS, which has an extra 15 amino acids between the initiator Met and the second amino acid, Lys, was expressed at a level of approximately 18% of total cell protein (approximately 50 mg/liter of bacterial culture). The fusion HisRS was purified to > 99% by a combination of anion exchange and cation exchange chromatography of the S100 fraction. The predicted MWs of the native and fusion proteins are 47,932 and 49,717, respectively. The mass of the active fusion HisRS was estimated to be 94,000 Da by Sephacryl S-200 gel filtration chromatography and 108,200 Da by nondenaturing PAGE. Both methods show that the functional enzyme is a dimer of two identical subunits. SDS-PAGE analysis of purified fusion HisRS with or without reduction showed a single band of M(r) = 53.7 kDa. PMID- 8248029 TI - Isolation of an immunosuppressive lectin from Phaseolus vulgaris L. cv Cacahuate using stroma. AB - An immunosuppressive lectin was isolated from seed of Phaseolus vulgaris cv Cacahuate using physically entrapped stroma. The lectin was found to be a 94 kDa tetrameric protein. When 50 micrograms, of this lectin were administered intraperitoneally 2 days before the immunization with sheep red blood cells, humoral response against the immunogen was completely inhibited. Other properties of the protein are discussed. PMID- 8248031 TI - 5th meeting of the European Placenta Group. Manchester, U.K., 8-11 September 1993. Abstracts. PMID- 8248030 TI - Improved chromatographic purification of peroxidase and beta-glucosidase from Hordeum vulgare seedlings. AB - Peroxidases (E.C. 1.11.1.7., hydrogen donor oxidoreductase) utilize hydrogen peroxide or substituted peroxides for the oxidation of a large number of substrates. Peroxidases are widely distributed and have been isolated from many higher plants (1). The wide distribution of the enzyme suggests that it could be of great biological importance, but the physiological functions and metabolic control of these enzymes are still poorly understood. The simultaneous presence of amine oxidase and peroxidase in cell walls suggests that the peroxide generated on oxidation of the amines could be utilized by the peroxidase (2,3). Recently we have purified an amine oxidase from Hordeum vulgare (4) and we have attempted to purify the peroxidase in order to study in vitro the reconstituted coupled system. beta-glucosidase (beta-D-glucoside glucohydrolase E.C. 3.2.1.21.) is capable of transforming glucosides in glucose and the corresponding aglycone or disaccharides as cellobiose, sophorose, gentiobiose. This enzyme is widely distributed in plants, fungi, bacteria, yeasts and animals (5,6). In the homogenate of Hordeum vulgare seedlings we also found beta-glucosidase activity and also attempted to purify beta-glucosidase. This enzyme copurified with peroxidase up to the last step. We report here the isolation of peroxidase and beta-glucosidase from Hordeum vulgare seedlings: some molecular and kinetic properties are given. PMID- 8248032 TI - Transferrin gene expression in maternal liver, fetal liver and placenta during pregnancy in the mouse. AB - Two clones that are homologous to the mouse liver transferrin gene were isolated from a differential screen performed on a mouse cDNA library constructed from placenta. Using an insert derived from the larger of these clones as a template for the generation of random primed cDNA probes, northern blots were conducted against total RNA collected sequentially from placenta (7 days before birth to birth), maternal liver (7 days before birth to birth) and fetal liver (5 days before birth to birth). An approximately 2.3 kb message was detected in all three tissues which was upregulated in late gestation. Message was very abundant in both maternal and fetal liver, and present, but weak, in placenta. The clones were partially sequenced and both clones contain sequence that is identical to mouse liver transferrin. The data presented demonstrate an increase in mRNA transferrin in late gestation in maternal and fetal liver. Additionally, the placenta expresses a gene homologous to liver transferrin and it also is upregulated in late gestation. PMID- 8248033 TI - Confocal immunofluorescence localization of collagen types I, III, IV, V and VI and their ultrastructural organization in term human fetal membranes. AB - The distribution of collagen types I, III, IV, V and VI in term human fetal membranes was examined using conventional and confocal indirect immunofluorescence techniques. Collagens I and III were present in most of the layers of fetal membranes except in the trophoblast layer contrary to what has been previously reported. Although collagen IV is considered to be a basement membrane component our study, using monoclonal and polyclonal antibodies, showed its consistent presence in the spongy and reticular layers in high intensity. This was first report on the distribution of type V collagen in the chorion where it was found in the reticular and in the trophoblast layers. Type VI collagen was present mainly in the amnion and the reticular layer. The ultrastructural examination of the extracellular matrix showed that the main fibrous skeleton of the fetal membranes was formed of large banded fibres (Ultrastructurally identical to collagens types I and III) connected together and to the epithelial basement membranes by networks of unbanded filaments (collagen types V, VI and other components). The extensive and continuous networks formed by these collagens may be a major factor responsible for the mechanical integrity of the fetal membranes. PMID- 8248034 TI - Characteristic alteration in the concentration of IV 3NeuAc alpha-nLc4Cer in the villi of human placenta during the gestational period. AB - A study on the ganglioside composition in the villi isolated from human placenta at various gestational periods was carried out by conventional procedures including thin-layer chromatography (TLC), TLC-immunostaining, negative ion fast atom bombardment mass spectrometry (FABMS) and exoglycosidase treatment. The major gangliosides in the villi were II3 NeuAc-LacCer (GM3) and IV3NeuAc alpha nLc4Cer, comprising 60-70% of the total gangliosides. The concentration of IV3NeuAc alpha-nLc4Cer per gram dry weight of tissue in the villi was found to be gradually decreased from the early to the late gestational period and the molecule with 2-hydroxy fatty acids was undetectable after 20 weeks of the gestational period. However, no significant correlation between the concentration of GM3 and the gestational periods was observed. Thus the characteristic alteration in the concentration of IV3NauAc alpha-nLc4Cer in the villi might be related to various functions of human placental villi during the gestational period. PMID- 8248035 TI - Epidermal growth factor (EGF) replaces estradiol for the initiation of embryo implantation in the hypophysectomized rat. AB - Several studies have provided evidence that epidermal growth factor (EGF) or transforming growth factor alpha (TGF alpha) mediate some of the physiological effects of estrogen. The object of the present study was to determine whether exogenous EGF could initiate embryo implantation in the rat, a response known to depend upon the action of estrogen in a progesterone-primed uterus. Immunocytochemical examination showed the presence of immunoreactive EGF, TGF alpha and the EGF receptor in luminal glandular epithelium of uteri on days 4,5 and 6 of pregnancy. EGF receptor was also present in the implanting embryo and in decidual cells of the uterine stroma. Attempts to initiate implantation by intravenous injection of murine EGF into ovariectomized or hypophysectomized delayed-implanting rats maintained with progesterone were unsuccessful. Implantation sites were found, however, in 24 of 33 (73 per cent) hypophysectomized progesterone-primed rats given 100 micrograms EGF immediately after intrauterine transfer of blastocysts from hypophysectomized delayed implanting animals. Although EGF is capable of initiating implantation in the delayed implanting rat model it remains to be determined whether this growth factor is responsible for the implantation inducing action of estrogen in vivo. PMID- 8248036 TI - Some structural studies on the galactose-containing polysaccharide from bovine placenta. AB - Polysaccharides were extracted from 8-month-old placenta with aqueous HgCl2. The protein-free material was purified by selective precipitation with Cetavlon in the presence of sodium borate at pH 8.5 and was homogeneous on molecular-sieve chromatography, electrophoresis, and on treatment with Concanavalin A. The preparation contained galactose and glucose as principal monosaccharides with 5 per cent of hexosamines. Methylation studies suggested that D-gluco and D galactopyranosyl units may be constituents of glucan and galactan respectively which form a molecular aggregate that does not dissociate during the fractionation procedures. After treatment of the fraction with beta-amylase, the proportion of glucose in the polysaccharide diminished, indicating the presence of (1-->4)-linked alpha-D-glucopyranosyl residues. Also, when the fraction was treated with a crude protease having glucosidase activity a residual alpha-D galactopyranan was isolated and found to contain non-reducing end-groups (30.0 per cent), 3-O-(39.5 per cent) and 3,6-di-O-substituted (30.5 per cent) units. The structure of the galactan was not modified according to methylation data, on removal of the glucosyl component. The polysaccharide fraction (pH 8.5 Cetavlon), isolated from bovine placenta, thus contains a glycogen-like material associated with a galactan as molecular aggregate. This galactan has not been previously recognized in bovine placenta and its occurrence in this organ supports the hypothesis that galactose-containing polysaccharides are involved in foetal development. PMID- 8248037 TI - Partial purification and characterization of the human placental serotonin transporter. AB - The human placental serotonin transporter was solubilized from purified brush border membranes using digitonin as the solubilizing agent. The solubilizate was subjected to wheat germ agglutinin-Sepharose 6B column chromatography, Centricon 100 ultrafiltration and Sepharose 6B gel filtration to yield a partially purified preparation of the serotonin transporter. Specific binding of the high affinity ligand paroxetine was used to monitor the transporter during the solubilization and the purification steps. The enrichment of paroxetine binding in the final preparation was 51-fold compared to the intact brush border membranes, taking into account the inactivation that occurred during purification. The partially purified transporter exhibited paroxetine binding characteristics which were similar to those of the transporter in intact membranes. The transporter in the partially purified preparation bound paroxetine with a high affinity (dissociation constant, 0.21 nM). The binding was inhibitable by serotonin but not by other monoamines, dopamine and norepinephrine, nor by the serotonin precursor 5-hydroxytryptophan. The antidepressants, imipramine, fluoxetine and desipramine inhibited the binding with a rank order of potency of imipramine = fluoxetine > desipramine. The approximate molecular weight of the transporter was assessed by molecular sieve chromatography on Sepharose 6B and was found to be 300,000. When reconstituted into proteoliposomes, the partially purified transporter was able to catalyse NaCl-dependent serotonin transport in these proteoliposomes. The results of this study show that the human placental serotonin transporter can be solubilized, partially purified and reconstituted in a transport-competent form and, in addition, provide some insight into the protein nature of the transporter. PMID- 8248038 TI - In vitro differentiation and ultrastructure of human extravillous trophoblast (EVT) cells. AB - Tissue explants of anchoring villi from the first trimester placentae cultured on extracellular matrix (Matrigel) give rise to EVT cells in vitro. This study was designed to address two issues important for further application of the described in vitro model: first, were the observed EVT cells derived by cell proliferation in vitro and second, what is the degree of homology between the in vivo and the in vitro differentiated EVT cells. The cultures (tissue and matrix) were prepared for light and electron microscopic (EM) examinations. Semi-thin sections from Spurr epoxy resin-embedded tissue were used to 'pop-off' the selected area for EM examination. Cell proliferation in vitro was assessed immunohistochemically using proliferative cell nuclear antigen (PCNA) antibodies. Since positive hPL immunostaining has been consistently demonstrated in the invasive subpopulation of EVT cells from placental bed in situ, hPL staining was used as a marker of EVT cell differentiation in vitro. It has been demonstrated that PCNA antibodies immunostained nuclei of cytotrophoblast cells from cell column at the base of the anchoring villi, indicating that these cells expressed proliferative activity in vitro. Cytotrophoblast proliferation resulted in the formation of the flattened zone of cell outgrowths at the tip of anchoring villi. Cells from the distal layer of the cell column detached gradually and migrated into the surrounding matrix. These cells appeared as individual, round-shaped EVT cells with smooth surface cell membrane. Their cytoplasm was rich in glycogen and contained large lipid droplets and flattened cisternae of the RER. Positive PCNA immunostaining, along with the presence of mitotic figures, indicated that EVT cells in vitro retained the ability for cell proliferation. As a result of cell proliferation and migration, the number of EVT cells increased during the culture period of 4 days. EVT cell glycogen content and lipid stores decreased progressively as they migrated into the matrix. Individual EVT cells, as well as EVT cell clusters, became surrounded by the clear zone of digested matrix. Some cells started to express strong positive staining with hPL antibodies as soon as they had migrated outside the villous explant. By day 4 of culture, a small percentage of EVT cells (about 5-10%) ceased to migrate, firmly attached to the substratum and appeared as irregular shaped cells with filopodia-like projections. Their cytoplasm contained dilated cisternae of RER, a small number of glycogen granules and bundles of actin-like filaments located in the cytoplasm inside the plasma membrane.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8248039 TI - [Oncogenes and suppressor genes in breast cancer]. PMID- 8248040 TI - [Hepatic toxicity of antitubercular agents. Role of different drugs. 199 cases]. AB - One hundred and ninety-nine cases of hepatic injury related to antituberculous treatment were analysed in order to assess the role played by each drug and to look for predisposing factors. This is a retrospective study of 169 cases reported in the literature and 30 cases reported to the regional pharmacovigilance centre of Paris Saint-Antoine. The mortality rate was related to the dose of isoniazid: it was 43 percent with a daily dose higher than 300 mg, and 9 percent with a daily dose of 300 mg or less (P < 0.001). Hepatic injury appeared significantly earlier in the case of rifampicin combination: 11 weeks without rifampicin and 2 weeks with rifampicin (P < 0.01). The role of pyrazinamide was difficult to determine because isoniazid and pyrazinamide were always used in combination. The influence of a preexisting liver disease could not be evaluated because of the small number of cases reported (8 cases). Alcoholism did not increase the mortality rate. Our results confirm the dose dependent toxicity of isoniazid. Because of the short time elapsed before the apparent onset of hepatitis observed with the rifampicin combination, a close supervision of the patients should be exerted during the first weeks of treatment. PMID- 8248041 TI - [Hemophagocytic syndrome in HIV infection]. AB - This report describes the clinical spectrum and outcome of the hemophagocytic syndrome (HS) in 5 HIV infected patients. All 5 patients presented with fever, hepatomegaly and/or splenomegaly, confusion or coma and respiratory symptoms. Severe anemia was associated with thrombocytopenia and with neutropenia in 4 cases. Diffuse intravascular coagulopathy was present in 2 cases. Liver function tests were abnormal in three patients. The diagnosis of HS was made 2 to 12 weeks after the onset of symptoms and required in most patients repeated examinations of the bone-marrow, showing infiltration by histiocytes with prominent phagocytosis of blood cells. In one case this infiltration was not seen in the bone-marrow but only in the liver and the spleen. Varicella, mycobacterium infection, oesophageal candidiasis, Kaposi sarcoma were observed in the evolution of 3 patients. Anaplastic large cell Ki-1 lymphoma was present in one case. Four patients died as a result of complications of HS. The one patient with lymphoma survived. PMID- 8248042 TI - [Cardiac tamponade in severe hypothyroidism. A rare cause]. AB - Pericardial effusion frequently occurs in patients with hypothyroidism, and this fully justifies the use of echocardiography at the time of diagnosis and during the follow-up of hypothyroidism. Signs indicating that the pericardial effusion is poorly tolerated are rare, and the development of pericardial tamponade is exceptionally reported. A case of tamponade in an elderly woman with severe hypothyroidism is described here. Some physiopathological particularities are emphasized, together with the value of echocardiography for the diagnosis and that of pericardial drainage for the treatment. PMID- 8248043 TI - [Endoscopic treatment of uterine myoma]. AB - The technique and results of endoscopic resection of submucosal uterine myoma performed in a series of 42 cases are presented. Following hysterography and hysteroscopy, endoscopic resection is carried out through a dilated cervix and under glycol infusion. The technique is very simple and the duration of the operation never exceeds 30 minutes. The postoperative period is short, and the average length of stay in hospital varies from 24 to 48 hours. Resection of submucosal myoma usually enables treatment with progestins to be discontinued. Moreover, when the submucosal myoma is not very large and solitary this technique offers an alternative to hysterectomy. PMID- 8248044 TI - [Diagnosis of diffuse encephalopathies in adults with HIV infection. I]. AB - The diagnostic approach of focal central nervous system lesions in AIDS patients is now well established. In contrast, it is extremely difficult to determine the cause of diffuse encephalopathies, occurring frequently at the terminal stage of AIDS. Imaging is usually non specific and laboratory investigations are seldom contributive. In most cases, the aetiological diagnosis is provided by post mortem examination. In this first part of the study the authors deal with viral encephalitides which represent a classical and frequent cause of diffuse encephalopathy in AIDS. HIV encephalitis usually causes a progressive brain disease resulting in severe dementia; imaging may show diffuse leucoencephalopathy and/or cortico-subcortical atrophy. CMV encephalitis is often asymptomatic, discovered at autopsy; however, this diagnosis should be considered in patients with an encephalopathy of rapid onset, discrete signs of meningitis, symptoms of myelo-radiculitis, or a systemic CMV infection. Varicella-zoster virus encephalitis is not uncommon and may occur in the absence of characteristic rash. Infections by herpes simplex and measles viruses are exceptional. PMID- 8248045 TI - [Edema of the uvula during ventilation with nasal mask]. PMID- 8248046 TI - [Hyponitremia during voluntary intoxications]. PMID- 8248047 TI - [Efficacy of thalidomide against esophageal ulceration in HIV infection]. PMID- 8248048 TI - [Post-streptococcal acute glomerulonephritis in an 84-year old male patient with biclonal dysglobulinemia]. PMID- 8248049 TI - [Post-transfusion cutaneous vasculitis in myelodysplastic syndrome]. PMID- 8248050 TI - [Radiation-induced hemorrhagic proctitis. Treatment by laser]. PMID- 8248051 TI - [Toxoplasmic seroconversion during pregnancy. Value of polymerase chain reaction for early diagnosis of fetal infections]. PMID- 8248052 TI - [Prostatic adenocarcinoma with false elevation of creatinine phosphokinase MB isoenzyme]. PMID- 8248053 TI - [Cortical blindness revealing progressive multifocal leukoencephalitis in AIDS]. PMID- 8248054 TI - [Measles vaccination]. PMID- 8248055 TI - [Central and peripheral nervous system motor conduction rate in HIV infection]. AB - In order to assess the central motor pathways of HIV-infected patients motor evoked potentials were recorded on the upper limbs after transcranial and cervical magnetic stimulations and compared with peripheral conduction velocities. Motor evoked potentials were measured on both sides of 28 patients, mostly at the AIDS stage. Our results showed that although peripheral neuropathy was often found, central motor pathways did not appear, at the level they were tested, to be directly or indirectly damaged by the virus. On the contrary, some hyperexcitability seemed to be present. PMID- 8248056 TI - [Biliary scintigraphy. Application to the study of chronic cholestasis]. AB - In a sample population of 49 subjects (7 normal, 42 with various liver diseases), the parameters of the activity/time curve of trimethylbromo-iminodicetic acid (TBIDA) biliary scintigraphy were compared with the clearances of bromosulfophthalein (BSP) and indocyanine green (ICG). Correlation between T1/2 and P2 BSP slope was r = 0.50 (n = 33; P < 0.01). Correlation between Tmax TBIDA and fractional ICG clearance (P ICG) was r = 0.65 (n = 44; P < 0.001). In 23 cases of chronic cholestasis correlations remained significant (T1/2-P2 BSP: r = 0.53; n = 17; P = 0.02; Tmax-P ICG: r = 0.59; n = 17; P < 0.01). A prospective study of 11 cases of chronic intrahepatic cholestasis (primary biliary cirrhosis 8, primary sclerosing cirrhosis 3) showed that these two types of tests varied concordantly. Biliary scintigraphy, therefore, seems to be an accurate method to explore hepatocellular mass (degree of hepatic insufficiency) and cholestasis. The validation of biliary TBIDA scintigraphy as hepatobiliary functional exploration method and the possibility to study intrahepatic "regions of interest" defined a priori would make it possible to obtain a functional estimate of hepatic segments or lobes, for example before wide liver excision. PMID- 8248057 TI - [Progressive encephalopathy in antiphospholipid antibody syndrome]. AB - The most frequent neurological complications of antiphospholipid antibodies syndrome (APS) are cerebral infarctions. In cases with recurrent strokes this syndrome may be associated with multi infarct dementia. We report the case of a 40-year old man presenting as "primary" APS with multiple cerebral infarctions. This patient had insidiously and progressively developed a cognitive impairment predominant in memory. The progressive encephalopathy associated with APS is very different from multi infarct dementia and could be caused either by a vascular mechanism (vascular occlusion with fibrin thrombi or disseminated vasculitis) or by an immunological mechanism. PMID- 8248058 TI - [Neurologic complications of cardiac myxomas]. AB - Cardiac myxomas are rare tumours complicated by faintness or syncope usually occurring in upright position and due to obstruction of a cardiac valve orifice. Myxomas are also manifested by neurological deficits caused by myxomatous or fibrino-cruoric cerebral emboli. Intracranial aneurysms and metastases are exceptional. The simplest and most reliable diagnostic method is transthoracic two-dimensional cardiac echography. Treatment consists of surgical excision which has very low morbidity, mortality and recurrence rates. PMID- 8248059 TI - [Value of a simplified test using biosynthetic 1,34 parathormone in the exploration of pseudo-hypoparathyroidism]. PMID- 8248060 TI - [Duodenal ectopic pancreas revealed by digestive hemorrhage]. PMID- 8248061 TI - [Severe hematologic manifestations of folate deficiency during pregnancy]. PMID- 8248062 TI - [Heyde syndrome: a questionable concept]. PMID- 8248063 TI - [Toxoplasmosis during pregnancy]. PMID- 8248064 TI - [Role of celiosurgery in the diagnosis and the treatment of gynecologic cancers]. PMID- 8248065 TI - [Thrombocytosis of celiac disease in adults: a diagnostic and prognostic marker?]. AB - In 14 of 23 patients seen with coeliac disease thrombocytosis was present (range: 420,000 to 789,000 platelets per cubic mm) and was unrelated to iron deficiency or inflammatory syndrome. Among patients with thrombocytosis (group I), 6 had an associated autoimmune disease; this association was absent in patients without thrombocytosis (group II). There was no correlation between thrombocytosis and lymphocyte count, plasma IgA, IgG, IgM and fibrinogen levels, presence of HLA B8 antigen or histological stage. On the other hand, group I patients had a lower plasma level of albumin, phosphorus and folates. We conclude that thrombocytosis is useful in the assessment of patients with coeliac disease and reflects an enhanced activity of the disease. Moreover, the presence of thrombocytes in these patients' blood may indicate a major risk of associated autoimmune disease. PMID- 8248066 TI - [HIV infection. Risky behavior in drug addicts using intravenous drugs. Experience in the Alpes-Maritimes region]. AB - HIV risk behaviours among intravenous drug users (IVDUs) were studied in southeast France, where the prevalence of HIV infection is high. Data were collected from a self-reported questionnaire distributed to drug addicts admitted in the health care or social institutions of the Alpes Maritimes department during November 1991. Among the 195 IVDUs who answered the questionnaire, 142 (73 percent) were male and 53 (27 percent) were female. One hundred and seventeen (62 percent) were HIV positive, 21 (11 percent) had an unknown HIV status, and 51 (27 percent) where HIV negative. Significant differences in HIV risk behaviours were found between HIV negative, unknown HIV status and HIV positive IDVUs: 37 percent of unknown status and 47 percent of HIV negative IVDUs continued to lend their syringe, compared with less than 20 percent HIV positive IVDUs (P < 0.001). Conversely, 57 percent of unknown HIV status and 39 percent of HIV negative IVDUs borrowed used syringes in the previous 6 months, compared with 73 percent of HIV positive IVDUs (p < 0.001). Regarding sexual prevention behaviours of patients who had had sexual intercourse during the previous 6 months: less than 20 percent of HIV negative IVDUs used condoms regularly, as against 47 percent of HIV positive IVDUs (P < 0.001). More than 70 percent of HIV negative versus 46 percent of HIV positive reported sexual intercourse without condom (p < 0.001). Logistic regression showed that the systematic use of condom was associated with a reduction of needle sharing, which suggests that IVDUs who use condoms have the best perception of risk. But the only factor associated with an increase in condom use is a HIV positive serology. These results indicate that in southeast France more efforts should be devoted to specific programmes aimed at increasing the use of condoms in the IVDU population. PMID- 8248067 TI - [Generalized BCG infection after intravesical instillations of Calmette-Guerin bacillus]. AB - BCG has been disappointing as immunotherapy of numerous cancers, but it has been clinically successful in the intravesical treatment of bladder carcinomas sparing the muscle coat; it has indeed become the reference treatment for this type of cancer. However, complications are repeatedly reported, including generalized BCGitis. We report such a case with positive BCG culture. From the cases already published there emerges a homogeneous and often subacute clinical presentation suggestive of an ordinary pathogen. Bacteriology is not very helpful, even when recent techniques are used, and therefore the diagnosis rests on the context and, when samples are taken, on suggestive histological findings. To discuss the physiopathology of BCGitis--generalized immune reaction or multifocal BCG proliferation--is not useless since treatment depends on it. It is probable that these 2 mechanisms working together can be incriminated justifying the prescription of both antibiotics and corticosteroids. When this is done, the prognosis seems to be favourable in most patients. Yet a strict respect of contra indications and a very careful subsequent radiotherapy should reduce the risks. PMID- 8248068 TI - [Adhesion molecules of Leu-CAMs leukocytes]. AB - Adhesion molecules of leucocytes Leu-CAMs, also called beta-2 integrins, are heterodimeric glycoproteins which play a crucial role in interactions of leucocytes between themselves or with fundamental intercellular substances. They comprise 3 elements: LFA-1 (CD11a/CD18) expressed at the surface of leucocytes, and in particular lymphocytes, M01 or MAC-1 (CD11b/CD18), and p 150.95 (CD11c/CD18) expressed only on granulocytes, monocytes and macrophages. Their structure, function and regulation are studied. These 3 elements differ in the ligand(s) to which they adhere. LFA-1 intervenes in the adhesion of lymphocytes T and B and of cells presenting with the APC antigen; it therefore plays an important role in lymphoblast proliferation, T-cell cytotoxicity and immunoglobulin synthesis. M01 and p 150.95 intervene in the adhesion of particles and germs opsonized by serum complement, thereby playing a fundamental role in the body's defence against bacterial and fungal infections. They also intervene in the adhesion of leucocytes onto the vascular endothelium and in their migration through this vascular wall towards the inflammatory focus. The pathologies related to a congenital deficiency of these adhesion molecules or to the alterations they acquired are dealt with. The use of monoclonal antibodies directed against leucocyte adhesion in animal experiments opens new therapeutic vistas. PMID- 8248069 TI - [Intolerance to sulfonamides in HIV infected subjects. Toxic and allergic origin]. AB - Intolerance to sulfonamides is very frequent in HIV-infected subjects and 10 times more common than in the general population. There are 2 types of intolerance to sulfonamides: early reactions with urticaria or angioedema, which are IgE-dependent, and late reactions with febrile rash, which occur between the 6th and 12th days of treatment and represent the vast majority of allergic manifestations in HIV-infected subjects. Clinically, these reactions resemble serum sickness, but all physiopathological hypotheses point to toxic process. The degradation of sulfonamides has two different pathways: the N-acetylation pathway which is genetically determined and saturable, and the cytochrome P450 pathway which produces toxic hydroxylamine metabolites "detoxified" by glutathione. In HIV-infected subjects detoxication is thought to be incomplete due to an acquired deficiency of glutathione and probably increased in the presence of a slow acetylation profile. PMID- 8248070 TI - [Cutaneous necroses after subcutaneous injections of amikacin]. PMID- 8248071 TI - [Pasteurellosis in an HIV infected patient]. PMID- 8248072 TI - [Bone hydatidosis. Pseudosarcomatous aspect]. PMID- 8248073 TI - [Lung cancer associated with histiocytosis X. A new case]. PMID- 8248074 TI - [Absence of antiphospholipid antibodies in migraine]. PMID- 8248075 TI - [Double paraneoplastic syndrome in a case of adenocarcinoma of the endometrium associated with Sezary syndrome]. PMID- 8248076 TI - [Inter-human transmission of Pneumocystis carinii. Risk evaluation in onco hematology]. PMID- 8248077 TI - [Sarcoidosis, danger of exposure to sun]. PMID- 8248078 TI - [Spondylodiscitis, an unusual complication of colonoscopy]. PMID- 8248079 TI - [Can breast cancer be prevented?]. PMID- 8248080 TI - [Neuromeningeal listeriosis in adults. Clinical aspects and contribution of cotrimoxazole in monotherapy]. AB - A series of 28 patients suffering from neuromeningeal listeriosis is reported. This disease is consecutive to infection by Listeria monocytogenes--an ubiquitous and opportunistic Gram-positive bacillus--and has become a public health problem: its incidence is increasing and its prognosis is very severe despite the development of new bacteriological identification methods. Human beings are contaminated by food, which explains the frequent outbreaks of epidemics which are widely publicized, the infection being one of the consequences of the unprecedented development of the food industry and the cold food chain. The predominant clinical picture is one of non-specific meningoencephalitis. In about 50 percent of the cases the subjects infected are "immunodepressed" and/or more than 60 years' old. The diagnosis is difficult since the bacteriological identification is delayed (direct examination of the cerebrospinal fluid is rarely positive) and this fluid may be sterile (hence the value of blood cultures). A probability treatment therefore must be initiated before the diagnosis is confirmed if an unfavourable outcome is to be avoided. In Listeria monocytogenes infection cotrimoxazole administered alone seems to be a better antibacterial therapy than the reference ampicillin-aminoside combination. PMID- 8248081 TI - [Volumetric analysis of urinary erythrocytes in the etiological diagnosis of hematuria]. AB - Intravenous pyelography and cystography may fail to localize the origin of haematuria. Microhaematuria is known to be present in 2 to 10 percent of the general population, usually without pathological consequences. Study of red cell morphology by phase contrast microscopy is effective in distinguishing between "glomerular" (from renal tissue) and "non-glomerular" (from urinary tract) erythrocytes, but this technique is not currently available in all laboratories. Urinary blood cell volume analysis has been presented as a simple and automatic alternative method. We compared these two techniques in 100 cases of haematuria of various origins. The cut-off point between glomerular and non-glomerular erythrocytes was set at 71 fl. Phase contrast microscopy always confirmed the clinical and/or histological diagnosis, but volume analysis did not: mean erythrocyte volume of glomerular origin was 66.6 +/- 10.4 fl, while non glomerular volume was 94.5 +/- 17 fl (P < 0.001); cell volume analysis was confirmative in only 72 percent of all diagnoses (65 percent of microhaematurias, 83 percent of macrohaematurias); sensitivity was 65 percent and specificity 85 percent for glomerular erythrocytes. Due to poor performance, urinary red volume analysis is not an acceptable alternative method to phase contrast microscopy when searching for the site of bleeding. PMID- 8248082 TI - [Acute pancreatitis caused by foreign bodies. 2 cases]. AB - We report the cases of two patients with acute pancreatitis caused by the insertion of foreign bodies in the ampulla of Vater: vegetal residues in the first case and beltbuckle in the second case. The acute pancreatitis was revealed by severe epigastric pain, jaundice and a serum amylase increase to more than 9 times the upper limit of normal level. The presence of foreign bodies was detected by endoscopic retrograde cholangiography and pancreatography in the first case and by abdominal X-ray examination in the second case. Extraction of vegetal residues was performed by endoscopic route whereas that of the beltbuckle required surgery. From these 2 cases and 8 others previously reported cases, the clinical features and course of acute pancreatitis caused by foreign bodies have been analysed. PMID- 8248083 TI - [Atypical mycobacteria infections in AIDS. Role and diagnosis]. AB - Atypical mycobacteria are responsible for systemic infections which occur late in the course of AIDS. The Mycobacterium avium intracellulare complex is the most frequently blamed species. Clinical and laboratory data are not specific; they consist of protracted fever, gastrointestinal and respiratory disorders entangled with the numerous manifestations of the multiple pathologies observed in these patients. The diagnosis is ascertained by the isolation of mycobacteria from blood cultures (using the isolator lysis-centrifugation and the Bactec 6B system) and from tissue cultures (hepatic parenchyma, bone marrow, etc.). Pathological examinations of specimens obtained from liver needle biopsy, duodenal biopsy and myelogram are very useful in providing a more rapid diagnosis. PMID- 8248084 TI - [Non-enzymatic glycosylation of proteins. Complications of diabetes mellitus, aging and kidney failure]. AB - Non-enzymatic protein glycosylation is the first stage of the reaction described by L.C. Maillard. When the reaction progresses beyond that stage the long half life molecules are damaged by formation of intermolecular crosslinking. The recent discovery of pentosidine, a crosslink between lysine and arginine residues, has demonstrated that advanced Maillard reaction is accelerated in diabetic patients with severe complications. Moreover, high tissue and plasma levels of pentosidine have been found in uraemia. The formation of advanced Maillard end-products (AGE) in plasma proteins constitutes a source of cell stimulation which induces macrophages to secrete cytokines, interleukin-1 and tumoral necrosis factor. Similarly, endothelial cells are induced to increase the permeability and production of the procoagulant factor. These mechanisms are thought to play an important role in the pathogenesis of atherosclerosis, nephropathy and thromboembolic disorders of diabetes. The discovery of beneficial effects of amino-guanidine, an inhibitor of advanced Maillard reaction, in the prevention of experimental diabetic complications opens a new line of investigation and new hopes for diabetics. PMID- 8248085 TI - [Gelatinous bone marrow necrosis in anorexia nervosa]. PMID- 8248086 TI - [Arterial hypertension, hypokalemia and adrenal mass. Beware of hasty etiological conclusions]. PMID- 8248087 TI - [Pulmonary lymphangiomyomatosis. Early diagnosis by computerized tomography and video-thoracoscopy]. PMID- 8248088 TI - [Angioneurotic edema acquired during chronic lymphoid leukemia. A case]. PMID- 8248089 TI - [Tetraplegia following assisted ventilation for status asthmaticus. Neuropathy or myopathy?]. PMID- 8248090 TI - Solution conformation of a synthetic bis-headed inhibitor of trypsin and carboxypeptidase A: new structural alignment between the squash inhibitors and the potato carboxypeptidase inhibitor. AB - The trypsin carboxypeptidase peptide inhibitor (TCPI) which inhibits both trypsin and carboxypeptidase A has been chemically engineered by modification of the Ecballium elaterium trypsin inhibitor II (EETI-II). The solution conformation of TCPI, studied by two-dimensional nuclear magnetic resonance, was shown to be very close to those of squash inhibitors. Only limited deviations of the trypsin binding loop compared to its location in the EETI-II/trypsin complex were detected. It was also shown that the position of the C-terminal tail did not significantly change from the position observed in the complex between carboxypeptidase A and the potato carboxypeptidase inhibitor (PCI). The conformation of TCPI was carefully compared with the PCI one and a new structural alignment between the two microproteins is proposed. This alignment points out the very good conservation in the two inhibitors of a subdomain comprising segments 7-15, 19-22 and 25-28. Most importantly, the 2-19 disulfide bridge of TCPI was not structurally conserved in PCI and appeared to be rather unimportant for the early folding process of these molecules. This result agrees with the recent observation that the 2-19 bridge is the last to be formed in the folding of the squash inhibitor EETI-II and suggests that this is also the case during the folding of the potato carboxypeptidase inhibitor. PMID- 8248091 TI - Mix'n'Match: an improved multiple sequence alignment procedure for distantly related proteins using secondary structure predictions, designed to be independent of the choice of gap penalty and scoring matrix. AB - A new multiple sequence alignment procedure is presented. Several different multiple alignments are made using differing criteria. Having divided the sequences into strongly conserved regions (SCRs) and loosely conserved regions (LCRs), the 'best' alignment for each LCR is chosen, independently of the other LCRs, from a selection of possibilities in the multiple alignments. To help make this choice for each LCR, the secondary structure is predicted and shown alongside each different possible alignment. One advantage of this method over automatic, non-interactive methods, is that the final alignment is not dependent on the choice of a single set of scoring parameters. Another is that, by allowing interactive choice and by taking account of secondary structural information, the final alignment is based more on biological rather than mathematical factors. This method can produce better alignments than any of the initial automatic multiple alignment methods used. PMID- 8248092 TI - Molecular dynamics of the 5-HT1a receptor and ligands. AB - A 3-D model of the human 5-HT1a receptor was constructed from its amino acid sequence by computer graphics techniques, molecular mechanics calculations and molecular dynamics simulations. The model has seven alpha-helical membrane spanning segments, which form a central core containing a putative ligand binding site. Electrostatic potentials 1.4 A outside the water accessible surface were mainly negative on the synaptic side of the receptor model and at the postulated ligand binding site, and positive in the cytoplasmic domains. The negative electrostatic potentials around the synaptic domains indicate that positively charged ligands are attracted to the receptor by electrostatic forces. Molecular dynamics simulations of the receptor model with serotonin, ipsapirone, R(-) methiothepin or S(+)-methiothepin in the central core suggested that up to 22 different amino acid residues may form a ligand binding pocket, and contribute to the specificity of ligand recognition and binding. PMID- 8248093 TI - Modeling the uncleaved serpin antichymotrypsin and its chymotrypsin complex. AB - In order to provide a structural reference for protein engineering experiments involving the serpin alpha 1-antichymotrypsin (ACT) and its complexes with chymotrypsin and DNA, a homology model of ACT has been constructed based on the 3 D structure of the related protein ovalbumin [29% identical and 44% similar; see Stein, P., Leslie, A., Finch, J., Turnell, W., McLaughlin, P. and Carrell, R. (1990) Nature, 347, 99-102]. After mapping the amino acid sequence of ACT onto the peptide backbone of ovalbumin, the resulting model was subjected to simulated annealing and energy minimization. Overall, the final ACT model is structurally similar to ovalbumin, although the 2.4 A root mean square deviation of corresponding C alpha atoms reflects the presence of regions exhibiting notable structural differences. The hydrogen bond stereochemistry of the ACT model is consistent with patterns found in high resolution protein structures and 92% of its backbone atoms have acceptable conformations when evaluated in a Ramachandran analysis. Significantly, the homology model serves as a structural reference for protein engineering experiments aimed at redesigning the functional properties of ACT, particularly with regard to its protease-bound conformation. Additionally, the homology model may be useful as a probe for solving the crystal structures of certain ACT variants (e.g. Thr345-->Arg) by molecular replacement methods. Ultimately, the homology approach may be applied toward the construction of other serpin models starting with an experimentally determined structure of uncleaved ACT as a template. PMID- 8248094 TI - A study of structural determinants in the interleukin-1 fold. AB - The structures of interleukin-1 beta, basic fibroblast growth factor and Erythrina trypsin inhibitor have been analysed in order to determine whether the hydrophobic core remains conserved, even when the structures have extremely low sequence similarities. We find that there are significant differences in the way each protein achieves a satisfactory arrangement of core residues and that positions which contribute to the core of one structure are not guaranteed to contribute to the integrity of another. Furthermore, the side-chain packing arrangements of these core residues vary significantly between the three structures. During this analysis the side-chain rotamers for three independently determined interleukin-1 beta structures were also compared. It was found that although buried residues are generally in agreement the remaining residues frequently occupy different rotamers in the three structures. This suggests that although meaningful studies are possible for buried side-chains the results obtained from equivalent analyses of accessible residues should be treated with caution. These results are discussed with specific reference to the optimization of side-chain packing in proteins of known structure. PMID- 8248095 TI - The fuzzy-end elimination theorem: correctly implementing the side chain placement algorithm based on the dead-end elimination theorem. AB - Recently it has been shown that the dead-end elimination theorem is a powerful tool in the search for the global minimum energy conformation (GMEC) of a large collection of protein side chains given known backbone coordinates and a library of allowed side chain conformational states, also known as rotamers. A side chain placement algorithm based on this theorem iteratively applies this theorem to single as well as to pairs of rotamers leading to the identification of rotamers, single or pairs, that are incompatible with the GMEC and that can thus be qualified as 'dead-ending'. Here we formulate a theorem which proves that contrary to intuition dead-end rotamer pairs cannot simply be discarded from consideration in the iterative process leading to the further elimination of dead end rotamers. We refer to this theorem as the fuzzy-end elimination theorem. We also describe how the obtained dead-end rotamer pairs can contribute to the search for the GMEC in the protein side chain placement problem. Hence the present work forms a theoretical basis for the correct implementation of a side chain placement algorithm based on the dead-end elimination theorem. In addition, possible future perspectives are presented. PMID- 8248096 TI - Breakdown in the relationship between thermal and thermodynamic stability in an interleukin-1 beta point mutant modified in a surface loop. AB - Sequence variants of the beta-barrel protein interleukin-1 beta have been analyzed for their stabilities toward irreversible thermal inactivation by monitoring the generation of light scattering aggregates on heating. The derived temperatures for the onset of aggregation (T(agg) values) correlate well with the free energies of unfolding of these proteins with the exception of one variant, Lys97-->Val (K97V), which undergoes aggregation at a temperature 7 degrees C lower than expected based on its thermodynamic stability. This lower than expected thermal stability may be due to generation of an aggregation-prone unfolding intermediate at a temperature lower than the Tm of the global transition. This hypothesis is supported by the location of residue 97 in the long 86-99 loop which has structural features suggesting it may comprise a small, independent folding unit or microdomain. The excellent correlation of thermal and thermodynamic stabilities of seven of the eight variants tested is consistent with accepted models for thermal inactivation of proteins. At the same time the poor fit of the K97V variant underscores the risk in using thermal stability data in quantitative analysis of mutational studies of the folding stability of proteins. PMID- 8248097 TI - Trp59 to Tyr substitution enhances the catalytic activity of RNase T1 and of the Tyr to Trp variants in positions 24, 42 and 45. AB - Using point mutated overproducing strains of E. coli, ribonuclease T1 was prepared with the single substitutions Tyr24Trp, Tyr42Trp, Tyr45Trp or Trp59Tyr and the corresponding double substitutions Tyr24Trp/Trp59Tyr, Tyr42Trp/Trp59Tyr and Tyr45Trp/Trp59Tyr. Steady state kinetics of the transesterification reaction for the two dinucleoside monophosphate substrates guanylyl-3',5'-cytidine and guanylyl-3',5'-adenosine indicate that the tryptophan can be introduced in different positions within the ribonuclease T1 molecule without abolishing enzymatic activity. The Trp59Tyr exchange even enhances catalysis of the cleavage reaction (kcat/Km) relative to the wild type enzyme and similar effects are found with single tyrosine to tryptophan substitutions. For the pH dependencies of the guanylyl-3',5'-cytidine transesterification reaction of wild type ribonuclease T1 and of the variants, typically bell-shaped curves are observed with a plateau in the range pH 4.5-7.0. Their shapes and slopes indicate that the enzymes are comparable in their macroscopic pKa values. At pH 7.5, the variant Tyr45Trp/Trp59Tyr shows a more than 3-fold higher transesterification activity for guanylyl-3',5'-adenosine and a 2-fold increase for guanylyl-3',5'-cytidine compared to the wild type enzyme, i.e. this variant catalyses the transesterification of the substrate guanylyl-3',5'-adenosine with the same or better efficiency as guanylyl-3',5'-cytidine. PMID- 8248098 TI - Strong inhibition of fibrinogen binding to platelet receptor alpha IIb beta 3 by RGD sequences installed into a presentation scaffold. AB - In order to probe the structural constraints on binding of RGD sequences to the platelet receptor alpha IIb beta 3 we have used recombinant DNA techniques to install the RGD sequence into 'presentation scaffolds', small proteins of known 3 D structure chosen to present guest sequences in constrained orientations. Using Escherichia coli expression systems we made sequence variants in which loop residues of the immunoglobulin VL domain REI and of human interleukin-1 beta were replaced (without changing polypeptide length) by the RGD sequence at positions predicted, based on small molecule studies, to orient the RGD moiety into an active conformation. These variants do not compete for fibrinogen binding to alpha IIb beta 3 up to almost 1 mM concentration. Unfolded or proteolytically fragmented forms of these same proteins do compete, however, showing that the RGD sequences in the mutants must be prohibited from binding by constraints imposed by scaffold structure. To suppress the effects of such structural constraints we constructed two sequence variants in which RGD-containing sequences 42-57 or 44 55 from the snake venom platelet antagonist kistrin were inserted (this increasing the length of the loop) into the third complementarity determining loop of REI. Both of these variants compete strongly for fibrinogen binding with IC50s in the nM range. These results, plus data on kistrin-related peptides also presented here, suggest that the molecular scaffold REI is capable of providing to an installed sequence a structural context and conformation beneficial to binding. The results also suggest that in order to bind well to alpha IIb beta 3, RGD sequences in protein ligands must either project significantly from the surface of the scaffold and/or retain a degree of conformational flexibility within the scaffold. Molecular scaffolds like REI should prove useful in the elucidation of structure-function relationships and the discovery of new active sequences, and may also serve as the basis for novel therapeutic agents. PMID- 8248099 TI - Construction of multiple copy of alpha-domain gene fragment of human liver metallothionein IA in tandem arrays and its expression in transgenic tobacco plants. AB - Metallothioneins (MT) are low molecular weight, cysteine-rich, metal-binding proteins. An MT molecule contains two domains which appear to act independently- an alpha-domain, which is characterized by cadmium-binding, and a beta-domain, which binds preferentially to copper. Based on this conception, DNA duplex encoding the alpha-domain (106 bp) of human MT-IA was constructed from a chemically-synthesized oligomer by repair synthesis and enzymatic ligation and cloned into pUC19. The genes cloned were sequenced and found to be in the correct order as designed. Synthetic directional adapters were attached to the terminals of the alpha-domain gene fragment of human MT-IA to establish complete control over fragment orientation during ligation. The use of these directional adapters thereby ensured the production of multiple copies of the alpha-domain in tandem arrays. The successive alpha-domains were linked by a peptide linker consisting of 10 residues. A chimeric gene containing 12 cloned tandemly repeated copies of the 106 bp alpha-domain DNA was introduced into tobacco cells on a disarmed Ti plasmid of Agrobacterium tumefaciens. A total of 10 different transgenic tobacco plants were generated, of which two showed root and shoot growth unaffected by up to 200 mg/l kanamycin and 100 microM cadmium, whereas root growth of control plants was severely inhibited and leaf chlorosis developed on media containing only 10 microM cadmium. PMID- 8248100 TI - Overproduction of bovine beta-casein in Escherichia coli and engineering of its main chymosin cleavage site. AB - A cDNA clone containing the entire coding region for bovine beta-casein A3 flanked by 53 base pairs of 5' non-coding and 358 base pairs of 3' non-coding sequences was isolated from a bovine mammary cDNA phagemid library. The coding segment for mature beta-casein was subcloned into the T7 expression system, in which the expression of recombinant beta-casein was controlled by the T7 gene 10 promoter and ribosome binding site. High level expression of Met-beta-casein to approximately 20% of the total soluble proteins was obtained in Escherichia coli within 2 h after induction of T7 RNA-polymerase synthesis. In an attempt to induce secretion the coding segment for mature beta-casein was coupled to the ompA translational initiation signal and signal peptide coding sequence but no secretion of the fusion protein and no processing of the signal peptide from the fusion protein was observed. Instead, the Met-beta-casein could be isolated in a soluble form from E.coli cells after an osmotic shock, indicative of a periplasmic location. This procedure did not lyse the cells. The protein was purified to homogeneity after a pH 4.8 isoelectric precipitation followed by reversed-phase high-performance liquid chromatography. The beta-casein cDNA was altered to change the main chymosin cleavage site in beta-casein at position 192 193 in two ways, namely from Leu-Tyr to Pro-Pro and to Leu-stop.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248101 TI - Expression of the catalytic subunit of cAMP-dependent protein kinase in Escherichia coli: multiple isozymes reflect different phosphorylation states. AB - The catalytic subunit of mouse cAMP-dependent protein kinase expressed in Escherichia coli was separated into three distinct species using Mono-S ion exchange chromatography. These isoenzymes corresponded to three isoelectric variants with pIs of 6.4 (30%), 7.2 (60%) and 8.2 (10%). The Stokes' radius of each form was 27.7, 27.1 and 26.3 A respectively. Using electrospray mass spectroscopy the differences between the isozymes were shown to be due to phosphorylation, with each form differing by 80 mass units corresponding to a single phosphate. The fully phosphorylated recombinant enzyme contained four phosphates while the dominant isozyme contained only three. Since the enzyme is not phosphorylated when active site mutations are introduced into the C-subunit, these phosphates are incorporated in an autocatalytic mechanism and are not due to E. coli protein kinases. When the recombinant enzyme was compared with the mammalian porcine heart enzyme significant differences in post-translational modifications were observed. The mammalian enzyme could also be separated into two isozymes. However, in contrast to the recombinant enzyme, the mammalian isozymes displayed an identical mass of 40 840. This correlated with two different post-translational modifications: two phosphates and an N-terminal myristyl moiety. The importance of post-translational modifications, and in particular the phosphorylation state, for the expression of eukaryotic proteins in E. coli is discussed. PMID- 8248102 TI - Expression of deletion constructs of bovine beta-1,4-galactosyltransferase in Escherichia coli: importance of Cys134 for its activity. AB - Bovine beta-1,4-galactosyltransferase (beta-1,4-GT; EC 2.4.1.90) belongs to the glycosyltransferase family and as such shares a general topology: an N-terminal cytoplasmic tail, a signal anchor followed by a stem region and a catalytic domain at the C-terminal end of the protein. cDNA constructs of the N-terminal deleted forms of beta-1,4-GT were prepared in pGEX-2T vector and expressed in E. coli as glutathione-S-transferase (GST) fusion proteins. Recombinant proteins accumulated within inclusion bodies as insoluble aggregates that were solubilized in 5 M guanidine HCl and required an 'oxido-shuffling' reagent for regeneration of the enzyme activity. The recombinant beta-1,4-GT, devoid of the GST domain, has 30-85% of the sp. act. of bovine milk beta-1,4-GT with apparent Kms for N acetylglucosamine and UDP-galactose similar to those of milk enzyme. Deletion analyses show that both beta-1,4-GT and lactose synthetase activities remain intact even in the absence of the first 129 residues (pGT-d129). The activities are lost when either deletions extend up to residue 142 (pGT-d142) or Cys134 is mutated to Ser (pGT-d129C134S). These results suggest that the formation of a disulfide bond involving Cys134 holds the protein in a conformation that is required for enzymatic activity. PMID- 8248103 TI - Production and properties of a factor X-cellulose-binding domain fusion protein. AB - A fusion protein, FX-CBDCex, which comprises factor X with a cellulose-binding domain (CBDCex) fused to its C-terminus, was produced in BHK cells. It was purified from the culture medium by affinity chromatography on cellulose. FX CBDCex could be activated to FXa-CBDCex with Russell viper venom. FXa-CBDCex was as active as FXa against a chromogenic substrate and against proteins containing the Ile-Glu-Gly-Arg sequence hydrolysed by FXa. FXa-CBDCex retained its activity when adsorbed to cellulose. PMID- 8248104 TI - Asbestos, asbestosis, and lung cancer. PMID- 8248105 TI - Gastritis associated with Gastrospirillum hominis in children. Comparison with Helicobacter pylori and review of the literature. AB - Interest in possible microbiological causes of gastritis has significantly increased since the discovery of Helicobacter pylori. Recently a spiral bacterium named Gastrospirillum hominis was described in association with chronic gastritis in adult patients. Here, we present the finding of Gastrospirillum hominis in the gastric biopsies of two children who underwent upper endoscopy for gastrointestinal symptoms. The frequency of Gastrospirillum hominis (0.3%) in our pediatric population was similar to that reported in adults. We observed a chronic gastritis associated with the spiral bacteria which was milder than the gastritis noted in our pediatric patients with Helicobacter pylori infection. Further comparisons between these two organisms, as well as the literature on Gastrospirillum hominis, are also reviewed. PMID- 8248106 TI - Signet ring variant of lobular carcinoma of the breast: a clinicopathologic and immunohistochemical study. AB - Signet ring carcinoma of the breast often metastasizes to gastrointestinal tract and female genital tract. We report clinicopathologic features of 10 breast carcinomas with signet ring features, five of which had unusual metastatic patterns. The primary breast tumor in all these cases was lobular carcinoma. Although signet ring cells were prominent in metastatic sites, the primary tumor lacked signet ring cells in two cases. A linitis plastica-like presentation and presence of signet ring cells in gastric metastases raised a strong possibility of primary gastric carcinoma in three cases. The monoclonal antibody to gross cystic disease fluid protein (GCDFP-15) was positive in signet ring cell-rich areas in the primary breast tumor (8/10) and/or in the metastases in all cases. For comparison we studied GCDFP-15 immunoreactivity in 10 infiltrating lobular and 10 infiltrating ductal breast carcinomas with no obvious signet ring cells, and in 14 signet ring carcinomas from other sites (10 gastric, 2 prostatic, 2 colonic). The gastric, colonic, and one prostatic signet ring carcinoma were nonreactive. One prostatic signet ring carcinoma exhibited focal but unequivocal positivity with GCDFP-15. The cases of this report reinforce the concept that signet ring carcinoma of the breast is usually a variant of lobular carcinoma and not a distinct entity. Signet ring cell predominance in metastases, even in the absence of signet ring cells in the primary tumor, attest to the morpho functional heterogeneity of lobular carcinoma. GCDFP-15 is a sensitive marker for signet ring breast carcinoma and a very useful adjunct tool in the diagnosis of metastatic signet ring carcinoma of mammary origin. PMID- 8248107 TI - Frequent p53 overexpression in therapy related myelodysplastic syndromes and acute myeloid leukemias: an immunohistochemical study of bone marrow biopsies. AB - p53 overexpression was studied immunohistochemically in paraffin-embedded bone marrow biopsies using a recently described technique for antigen retrieval based on microwave oven heating of paraffin sections. Using a monoclonal antibody (PAb1801) that reacts with human cellular p53, nuclear staining was detected in 7/11 (63%) therapy-related myelodysplastic syndromes and in 3/4 (75%) therapy related acute myeloid leukemias. Conversely, staining for p53 was seen only in 9/40 (22%) cases of "primary" hematologic conditions (P < 0.007); these included myelodysplastic syndromes [#2], acute myeloid leukemia [#4], and chronic granulocytic leukemia in accelerated phase or blast crisis [#3]. Biopsies of normal controls and of chronic granulocytic leukemia in stable phase were consistently p53(-). Nine of the 10 karyotyped p53(+) acute myeloid leukemia/myelodysplastic syndrome cases showed complex cytogenetic findings with frequent involvement of chromosome 5 and/or 7. Only four of the 33 karyotyped p53(-) cases showed similar cytogenetic changes. Chromosome 17 involvement was present in four of 13 (31%) cytogenetically assessed p53+ cases, but in none of the p53(-). In univariate analysis, p53 expression in both MDS and AML was significantly associated with shorter survival. The frequent overexpression of p53 in therapy-related myelodysplastic syndromes, therapy-related acute myeloid leukemias and in accelerated phase/blast crisis, chronic granulocytic leukemia and its strong association with complex karyotypes suggests an important role of this gene in the pathogenesis of these leukemic conditions. PMID- 8248108 TI - Coexistent anti-neutrophil cytoplasmic antibody and antiglomerular basement membrane antibody associated disease = report of six cases. AB - From a series of 95 patients biopsied for rapidly progressive glomerulonephritis, twelve patients were identified with anti-glomerular basement membrane-mediated renal disease who were also tested for antineutrophil cytoplasmic antibody (ANCA). Six patients had both anti-glomerular basement membrane and ANCA antibodies. Three of the latter six patients had significant extrarenal disease, including severe hemoptysis, while the remaining three patients had only renal disease. The three patients with extrarenal disease had either a myeloperoxidase positive perinuclear-ANCA (two patients) or a proteinase-3-positive cytoplasmic ANCA (one patient). Two patients with renal disease alone had a myeloperoxidase negative and proteinase-3-negative perinuclear-ANCA, and one patient had a proteinase-positive cytoplasmic-ANCA. Renal biopsy in all six patients showed a severe necrotizing and crescentic glomerulonephritis involving 94 to 100% of glomeruli. Renal arteritis was also noted in one perinuclear-ANCA patient. Despite aggressive therapy with steroids, cyclophosphamide, and plasma exchange, two of the six double-antibody patients died and four are on dialysis. We conclude that ANCA is commonly present in anti-glomerular basement membrane associated disease and believe that this observation may have implications in the serologic evaluation of ANCA- and anti-glomerular basement membrane-positive patients. PMID- 8248109 TI - Correlation of histologic grade of breast carcinoma with cytologic features on fine-needle aspiration of the breast. AB - Histologic grade is an important prognostic factor for breast carcinoma. To determine whether histologic grade could be reproduced on fine-needle aspiration of the breast, we analyzed 50 cases of histologically proven infiltrating ductal carcinomas that had a preoperative fine-needle aspiration. The histologic grade was based on tubule formation, nuclear pleomorphism, and mitotic frequency [a modified Bloom-Richardson (BR) grading system]. Fine-needle aspirations were analyzed for cellularity, cellular arrangements, percentage of single cells, nuclear pleomorphism, mitoses, hyperchromasia, chromatin pattern, nucleoli, and background. The cases in BR Grades 1 and 2 (n = 28) were combined into one category for comparison with those in BR 3 (n = 22). BR 3 was more likely than BR 1 and 2 to be associated with marked nuclear pleomorphism (59 versus 18%, P < 0.05) and vesicular or coarse chromatin (77 versus 43%, P < 0.05). Of the 18 cases that showed both marked nuclear pleomorphism and either coarsely granular or vesicular chromatin on fine-needle aspiration, 13 (72%) were BR Grade 3 on histology. These 13 cases represent 59% of the 22 cases of BR Grade 3 on histology. Aspiration cytology is of some, but limited, use in predicting BR grade. As expected, nuclear features are most easily translated from histology to cytology. PMID- 8248110 TI - Premalignant nonepithelial lesions: a biological classification. AB - Premalignant lesions occur early in neoplastic development, are often small and multiple, lack one or more of the properties of cancers, often regress, and are often easily treatable. Such lesions may be derived from epithelial or nonepithelial cell populations. Many epithelial premalignant lesions can be identified by their confinement by a basement membrane. Mesenchymal, hematologic, and lymphoid premalignant lesions are difficult to recognize as most of these lesions have no defining morphologic features or anatomic boundary that identifies their premalignant nature. We have devised a classification for nonepithelial premalignant lesions, based on biologic behavior rather than morphology, that includes neoplastic lesions that have some but not all of the properties generally attributed to developed cancers. The five categories of nonepithelial premalignant lesions are: (a) lesions delimited by a basement membrane (e.g., melanoma in situ); (b) nonepithelial neoplasms having a low rate of metastasis (e.g., giant cell tumor of bone); (c) early forms of nonepithelial malignancy (e.g., Kaposi's sarcoma, patch stage); (d) indolent neoplasms that regularly progress to malignancies (e.g., chronic lymphocytic leukemia); and (e) certain low-grade neoplasms derived from germ cells (e.g., Grade 1 immature teratoma). Formal classification of the nonepithelial premalignant lesions would allow the pathologist to diagnose these biologically distinct entities as something other than simply benign or malignant, and would direct tumor biologists to study the key molecular properties that control the premalignant phenotype. PMID- 8248111 TI - Detection of Epstein-Barr virus by polymerase chain reaction in transbronchial biopsies of lung transplant recipients: evidence of infection? AB - Thirty-nine paraffin-embedded transbronchial biopsies from ten lung transplant recipients were evaluated by the polymerase chain reaction for the presence of Epstein-Barr virus (EBV). Six of these patients had positive EBV serology. Nine biopsies from nine non-transplant patients with nonspecific interstitial pneumonitis were studied as a control group. Eight biopsies from four of the ten transplant patients (40%) were positive for EBV by polymerase chain reaction, utilizing 40 cycles and two sets of primers (501/502) complementary to sequences within the Bam H1 W region of the viral genome. Five of these eight positive biopsies had been diagnosed as mild acute cellular rejection based on clinicopathologic criteria. Three of the four EBV-positive patients showed clinical improvement with antiviral agents prescribed for concomitant cytomegalovirus infection. Our data demonstrate the potential of polymerase chain reaction to detect small quantities of EBV in transbronchial biopsies from lung transplant recipients. Such findings should be interpreted cautiously in the evaluation of lung transplant recipients, since EBV can be detected in the absence of a lymphoproliferative disorder or an active pneumonitis caused by this virus. PMID- 8248112 TI - Myocardial risk factors other than human immunodeficiency virus infection may contribute to histologic cardiomyopathic changes in acquired immune deficiency syndrome. AB - This study was designed to evaluate whether myocardial risk factors other than those strictly related to human immunodeficiency virus infection contribute to histologic cardiomyopathic changes in acquired immunodeficiency syndrome patients. We analyzed 91 consecutive adult human immunodeficiency virus-positive autopsy cases (85% acquired immunodeficiency syndrome by Centers of Disease Control criteria) from 1987-1991 for histologic cardiomyopathic changes (e.g. myocyte hypertrophy and myocardial fibrosis). We correlated the presence of cardiomyopathy with the following common myocardial risk factors: hypertension, coronary artery disease, alcoholism, diabetes mellitus, and valve disease. Forty percent of all cases had cardiomyopathy. Hypertension and coronary artery disease were both more common in the cardiomyopathy group (P < 0.05), compared with those human immunodeficiency virus-positive cases without cardiomyopathy. The other myocardial risk factors did not differ significantly between the two groups when compared individually, but when these data were pooled, 67% of cardiomyopathic patients had one or more myocardial risk factors versus 45% in the noncardiomyopathic group (P < 0.05). Cardiomyopathic patients were also more likely to have multiple myocardial risk factors (P < 0.05). Nineteen percent of cardiomyopathic patients had myocarditis versus 11% in the noncardiomyopathic group (P = NS). Patient age, gender, risk factors for human immunodeficiency virus infection (71% intravenous drugs), and history or autopsy findings of viral infection (e.g. cytomegalovirus) did not differ significantly between the two groups. In our patient population, which is heavily weighted towards intravenous drug use, myocardial risk factors other than human immunodeficiency virus are common, and appear to be major contributors to histologic cardiomyopathic changes that might otherwise be attributed to human immunodeficiency virus infection alone. PMID- 8248113 TI - Quantitative morphologic assessment of nuclei extracted from paraffin for DNA flow cytometry. AB - We present a method for quality control of flow cytometric DNA content studies using nuclei extracted from paraffin-embedded tissue. This method is based on a quantitative morphologic assessment of extracted nuclei. Cell nuclei prepared from 22 paraffin-embedded tumors known to contain discrete diploid and aneuploid stemlines were deposited onto poly-L-lysine-coated glass slides. Nuclei were stained with Diff-quic and examined by light microscopy. Two hundred nuclei were counted and classified based on morphologic appearance into tumor and nontumor groups. Classification criteria included differences in nuclear size, nuclear chromatin structure, the degree of nuclear chromatin condensation, and the presence of nucleoli. Excellent agreement was found between two independent observers (R = 0.989) on the classification of nuclei. The relative number of tumor nuclei on the morphologic preparation was compared with the relative number of aneuploid cells in the DNA histogram. Good agreement was observed (R = 0.975) in all but three cases in which the relative number of tumor nuclei was underrepresented by the percentage of aneuploid nuclei in the DNA histogram. In each case, further analysis by image cytometry demonstrated a diploid and aneuploid component of the tumor cell population. This quantitative method of morphologic examination of the preparation ultimately analyzed by flow cytometry offers several distinct advantages, including: (a) identification of peaks in the DNA histogram, (b) assessment for selective loss of cell nuclei in the extraction process, and (c) identification of biologic heterogeneity in tumor populations. PMID- 8248114 TI - Papillary duct hyperplasia of the breast in children and young women. AB - Seventy-four patients who had papillary duct hyperplasia were studied to characterize the pathologic features of the lesions and to assess their precancerous significance. All but one were female. The median age at diagnosis was 17 yr with 52 (70%) between 15 and 25 yr, 18 (24%) < 15 yr and four (5%) 25 to 29 yr old. Age at follow-up ranged from 9 to 53 yr with 49 (66%) < 30 yr, 22 (30%) age 30 to 39 yr, and three (4%) age 40 yr or more. Median age at last follow-up was 28 yr. The most frequent presenting symptoms were a mass and/or nipple discharge. A family history of breast carcinoma was reported by 19 of 69 patients (28%) with available information. Three patterns of papillary duct hyperplasia were identified: sclerosing papilloma (31; 42%), papilloma (24; 32%) and papillomatosis (19; 26%). Prominent cysts, apocrine metaplasia, other benign proliferative changes, and mastitis that characterize juvenile papillomatosis were absent. All patients had an excisional biopsy followed by segmental mastectomy for two large lesions. Recurrences in the breast were detected in 12 (16%) of the patients after a median interval of 3 yr, more commonly in patients with sclerosing papilloma (5; 31%) and papilloma (5; 21%) than in those with papillomatosis (2; 11%). The risk of recurrence was not related to a family history of breast carcinoma. None of the patients developed mammary carcinoma or any other malignant neoplasm. These results indicate that children, adolescents, and young women with the types of papillary duct hyperplasia described in this report are not predisposed to develop breast carcinoma before age 30.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248115 TI - Epstein-Barr virus and primary lung lymphoma: a study utilizing the polymerase chain reaction. AB - Seven cases of primary lung lymphoma were analyzed for the presence of Epstein Barr virus DNA sequences by the polymerase chain reaction. The series included: four cases of diffuse, small lymphocytic lymphoma; one case of diffuse, intermediate lymphocytic lymphoma; one case of diffuse, small cleaved cell lymphoma; and one case of large cell, immunoblastic lymphoma. The latter occurred in a patient with acquired immunodeficiency syndrome. A 200-bp sequence of the Epstein-Barr nuclear antigen 1 gene was used as a template for PCR amplification. The only tumor that contained Epstein-Barr virus sequences was the immunoblastic lymphoma. These findings support previous observations that small lymphocytic lymphomas of the lung are not related to Epstein-Barr virus infection. In contrast, some large cell lymphomas may represent Epstein-Barr-virus--associated lymphoproliferative disorders. PMID- 8248116 TI - Blood group antigens in Barrett's esophagus and associated adenocarcinomas. AB - A panel of monoclonal antibodies (MoAbs) against BGAg A, B, and H and Lewis a, b, X, and Y (Le a, Le b, X, Y) was used to characterize and evaluate the blood group antigens (BGAg) immunohistochemistry in Barrett's esophagus (BE) and associated adenocarcinomas (ACA). Thirteen cases of ACA arising in metaplastic columnar epithelium of the distal esophagus were reviewed (12 men; 1 woman; mean age: 59). Twelve cases had juxtaposed BE of different types with low-grade dysplasia in five and high-grade dysplasia in three. ACAs were classified as well differentiated (one case), moderately differentiated (three cases), and poorly differentiated (four cases). Five cases had mixed features. Loss of BGAg H was recorded in four ACA and in one adjacent BE. Loss of BGAg B was noted in three ACAs, but in only one case in the adjacent BE. Incompatible expression of BGAg A and B was present in one case each. Anomalous expression of Le a was seen in seven ACAs and adjacent BE. Loss of Le b was noted in six ACAs. Deletion of X was seen in four ACAs and in BE. Loss of Y was present in the only nonsecretor patient. We conclude that significant alterations of BGAg occur in BE and in associated ACAs. Recognition of these antigenic alterations warrants further investigation to define a role for BGAg immunohistochemistry in the surveillance of BE patients at risk of developing ACA. PMID- 8248117 TI - The bone marrow agar section: a morphologic and immunohistochemical evaluation. AB - A technique for agar embedding of bone marrow aspirate particles is compared with the conventional aspirate smear and bone marrow biopsy by reviewing 503 consecutive bone marrow specimens. Immunohistochemical studies were performed on both agar sections and bone marrow biopsies on 43 paired specimens to compare the results between the two preparations. The results were also compared with traditional clot sections from ten control cases. Of the 382 cases with agar sections, 97.7% contained material in the agar that was diagnostic alone or supportive of the diagnosis made with the biopsy and aspirate smear. In two cases (0.4%), focal involvement by lymphoma was identified on the agar section but not in the biopsy sections or aspirate smears. The immunohistochemical studies showed superior immunoreactivity in agar sections by lymphoproliferative disorders when compared with bone marrow biopsy sections. Similar results between agar and conventional biopsy sections were found in cases of metastatic carcinoma and plasma cell dyscrasias. PMID- 8248118 TI - An image analysis workstation for the pathology laboratory. AB - Computer-based image analysis (IA) is a technology gaining importance in diagnostic pathology. Applications of IA in pathology include DNA ploidy analysis, quantitative immunohistochemistry, three-dimensional reconstruction of tissue sections, motility studies, and chromosomal analysis. Morphometry, the quantitative measurement of size, shape, and textural features of cells and tissues, is another rapidly developing area of IA in pathology. Morphometric IA allows the objective evaluation of subtle histologic and cytologic features to yield useful diagnostic and prognostic information. Research is currently underway to develop diagnostically useful applications of morphometric IA. Several image analysis workstations designed for the pathology laboratory are currently available. However, the high cost and software inflexibility of these instruments are prohibitive to many potential users limiting the practicality of IA and hindering research. We present a relatively inexpensive pathology IA workstation assembled from commercially available hardware and software components. System features and basic image processing methods are described. A variety of practical applications for the surgical pathology laboratory are illustrated, including spatial measurements of tumors, nerve and muscle biopsy evaluation, and nuclear morphometry for classification of lymphoid effusions and hepatocellular carcinoma. PMID- 8248119 TI - Spontaneous abortions and DNA ploidy. An application of flow cytometric DNA analysis in detection of non-diploidy in early abortions. AB - The value of DNA ploidy analysis utilizing paraffin-embedded materials in the detection of nondiploid gestations was examined in a year series of 217 spontaneous abortions in first trimester at a hospital. These placentas were histologically classified as follows: 19 hydropic placentas, six partial moles, three complete moles, and 189 nonhydropic placentas. Abnormal DNA content was found in 26 cases (12%). Twelve placentas (5.6%) were triploid, seven were tetraploid, and seven were nontriploid/tetraploid aneuploid. Of the triploid placentas, six were partial moles, one was a hydropic placenta, and the remaining five were nonhydropic. Since large mononuclear cells in villous mesenchyme were observed in the triploid and tetraploid as well as in the diploid hydropic placentas, in which category trisomic gestation was probable, these cells were not considered to be characteristic of trisomic placentas. Although placentas with hydropic villi had a significantly higher frequency of the nondiploid DNA content than nonhydropic placentas, with the exception of partial moles, no correlation between the histology of villi and DNA ploidy was observed. Flow cytometric analysis using paraffin-embedded materials appears to be of value for the retrospective study of spontaneous abortions, especially for assessing aneuploidy. PMID- 8248120 TI - Disseminated giant cell arteritis. AB - A 76-yr-old woman with widespread giant cell arteritis and polymyalgia rheumatica is described. The patient had an unusual simultaneous involvement of the cranial (temporal) arteries demonstrated by biopsy, and of large elastic arteries (aorta and its major branches), medium-sized and small muscular arteries, arterioles and vasa vasorum found at autopsy. Coronary arteritis was responsible for the fatal myocardial infarction. PMID- 8248121 TI - Orphan Annie nuclei and strawberry gall bladders. PMID- 8248122 TI - Episodic evolution of RNA viruses. PMID- 8248123 TI - Behavioral and magnetoencephalographic correlates of plasticity in the adult human brain. AB - Recent behavioral and physiological evidence suggests that even brief sensory deprivation can lead to the rapid emergence of new and functionally effective neural connections in the adult human brain. PMID- 8248124 TI - Hepatocyte nuclear factor 3/fork head or "winged helix" proteins: a family of transcription factors of diverse biologic function. AB - A family of transcription factors, first identified as hepatocyte nuclear factors (HNF-3 alpha, -3 beta, and -3 gamma) and as a homeotic Drosophila mutant, fork head, has been intensively studied for the past 4 years. Important findings have emerged about the structure of the DNA-binding portion of the proteins as well as biologic discoveries about the diversity of the family and its implied role in early development. PMID- 8248125 TI - Punctuated equilibrium and positive Darwinian evolution in vesicular stomatitis virus. AB - RNA viruses possess the potential for rapid evolution and serve as excellent models to test evolutionary theory. Molecular phylogenetic analysis of the P gene for a larger number of diverse natural isolates of vesicular stomatitis virus reveals no evidence for a molecular clock but instead shows a stepwise evolutionary pattern unlike that ever seen before. Each step out from the tree's ancestral root to terminal branch tips correlates not with time of virus isolation but with a south-to-north geographical progression from Panama to the United States. The grossly unequal rates of change within this single species imply an underlying mechanism at odds with the prevailing notion that neutral changes are the dominating feature of molecular evolution. This is also a demonstration of punctuated equilibrium at the molecular level. PMID- 8248126 TI - Multiple oligomeric states regulate the DNA binding of helix-loop-helix peptides. AB - To study the protein-protein interactions that allow Id, a negative regulator of cell differentiation, to inhibit the DNA-binding activities of MyoD and E47, we have synthesized peptides corresponding to the helix-loop-helix domains of MyoD, E47, and Id. We show that Id preferentially inhibits the sequence-specific DNA binding activity of MyoD, a muscle-specific protein, as compared to E47, a more ubiquitous protein. The Id helix-loop-helix domain itself forms stable tetramers, and its inhibitory activity arises from the formation of a heterotetrameric structure with MyoD. The formation of this higher order complex provides a general mechanism by which inhibitory proteins can generate sufficient interaction free energy to overcome the large DNA-binding free energy of dimeric DNA-binding proteins. PMID- 8248127 TI - Interaction of glucocorticosteroid receptor and wild-type or mutated 90-kDa heat shock protein coexpressed in baculovirus-infected Sf9 cells. AB - Coexpression of the human glucocorticosteroid receptor (hGR) and chicken 90-kDa heat shock protein alpha (chsp90) in recombinant baculovirus-infected Sf9 cells is a system that provides a large quantity of wild-type chsp90-hGR complexes able to bind hormone ([3H]triamcinolone acetonide; TA), sedimenting at 8 S, and displaceable to 11 S by BF4 and D7 alpha anti-chsp90 monoclonal antibodies. Thus, we were able to examine the effects of selective chsp90 mutations on hetero oligomeric complex formation. Two deletions involved hydrophilic regions, A between amino acids 221 and 290 and B between amino acids 530 and 581, and the third, Z, removed a central leucine heptad repeat region (amino acids 392-419). When these chsp90 mutants were expressed, the lack of displacement of [3H]TA receptor complexes on sucrose gradient by specific chsp90 antibodies was consistent with the formation of [3H]TA receptor complexes containing only endogenous insect hsp90. By using an immunoadsorption method and sedimentation analysis, we found that the deletion of region A precluded the interaction of chsp90 with the hGR, while B and Z deletions led to formation of abnormal complexes with the hGR, which displayed large forms (> 10 S), were unable to bind hormone, and apparently formed only small amounts of tightly bound nuclei hGR upon in vivo hormone treatment. As a whole, the data are consistent with distinct roles of hsp90 regions in hGR function. PMID- 8248128 TI - Specific enhancement of beta-adrenergic receptor kinase activity by defined G protein beta and gamma subunits. AB - The beta and gamma subunits of heterotrimeric guanine nucleotide-binding proteins (G proteins) have recently been shown to play an active role in signal transduction. Among other effects they enable translocation of the beta adrenergic receptor kinase (beta ARK) from the cytosol to the plasma membrane and thus permit phosphorylation and ultimately desensitization of beta-adrenergic receptors and other G-protein-coupled receptors. To investigate the specificity of this effect, we have purified various combinations of recombinant beta and gamma subunits expressed in Sf9 cells and measured their effects on beta ARK catalyzed phosphorylation of beta 2-adrenergic receptors and of rhodopsin. The combinations tested were beta 1 gamma 2, beta 1 gamma 3, beta 2 gamma 2, beta 2 gamma 3, and transducin beta gamma (beta 1 gamma 1). There were clear differences in enhancement of rhodopsin phosphorylation, with an order of efficacy beta 2 gamma 2 > beta 1 gamma 2 >> beta 2 gamma 3 approximately beta 1 gamma 3 approximately beta 1 gamma 1. The first two combinations had larger effects than a mixed beta gamma preparation from bovine brain. In enhancing phosphorylation of beta 2-adrenergic receptors, beta 1 gamma 2 was more efficient and potent than all other combinations. These data suggest a twofold specificity of beta gamma complexes in enhancing beta ARK-catalyzed receptor phosphorylation: the gamma subunits may be important in interacting with beta ARK, with gamma 2 being more potent than gamma 3, whereas the beta subunits may determine coupling to the receptors, with beta 2 being more effective than beta 1 for rhodopsin and beta 1 being more effective than beta 2 for beta 2-adrenergic receptors. PMID- 8248129 TI - Production and fluorescence-activated cell sorting of Escherichia coli expressing a functional antibody fragment on the external surface. AB - We have expressed a single chain Fv (scFv) antibody fragment, consisting of the variable heavy and variable light domains from two separate anti-digoxin monoclonal antibodies, on the external surface of Escherichia coli by fusing it to an Lpp-OmpA hybrid previously shown to direct heterologous proteins to the cell surface. This scFv fusion was expressed at a high level and was shown to bind the hapten with high affinity and specificity. Whole cell ELISAs, fluorescence microscopy, protease sensitivity, and flow cytometry all confirmed that the scFv was anchored on the outer membrane and was accessible on the surface. Utilizing fluorescence-activated cell sorting, we were able to specifically enrich scFv-producing cells from a 10(5)-fold excess of control cells in only two steps. The expression of antibody fragments on the surface of E. coli is being evaluated as an attractive method for the in vitro production and selection of useful antibody fragments. PMID- 8248130 TI - Functional expression and photoaffinity labeling of a cloned P2U purinergic receptor. AB - ATP and UTP can function as extracellular signaling molecules by activating plasma membrane receptors termed P2 purinergic receptors. In the present study a P2U receptor cDNA has been expressed in K-562 human leukemia cells, one of the few available mammalian cell lines that lacks an endogenous P2U receptor. In stably transfected cells, low micromolar concentrations of ATP or UTP activated the receptor, resulting in the mobilization of intracellular calcium but not the influx of extracellular calcium. A photoaffinity agonist of the P2U receptor, 3' O-(4-benzoylbenzoyl)adenosine 5'-[alpha-32P]triphosphate ([alpha-32P]BzATP), photolabeled several proteins in plasma membranes from the stable transfectant or from untransfected K-562 cells. The photolabeling of a 53-kDa protein was significantly greater in plasma membranes from the stable transfectant than from untransfected cells. A mutant receptor containing six consecutive histidine residues at its carboxyl terminus was constructed and used to verify that this 53 kDa protein was the P2U receptor. In plasma membranes from cells expressing the histidine-tagged P2U receptor, but not from cells expressing the wild-type receptor, a single [alpha-32P]BzATP-labeled protein with a molecular mass of 53 kDa was retained on a Ni(2+)-charged Sepharose column, which binds many proteins containing a polyhistidine tag. Photolabeling of the 53-kDa protein by [alpha 32P]BzATP was inhibited by ATP but not by UTP, raising the possibility that the P2U receptor may have distinct binding sites for each nucleotide. PMID- 8248131 TI - A method for producing monoclonal antibodies to human T-cell-receptor beta-chain variable regions. AB - Study of the T-cell repertoire in humans has been hampered by the lack of monoclonal antibodies (mAbs) to the T-cell receptor (TCR) variable region (V) gene products. We describe a method for producing mAbs to the human TCR beta chain V (V beta) gene products in which mice were immunized with a rat basophil cell line (RBL-2H3) transfected with the extracellular domain of the TCR heterodimer fused to the lambda chain of CD3. These cells acted as excellent immunogens for raising anti-TCR mAb and also formed the basis of a rapid screening assay. We generated mAbs against V beta protein of the TCR, showed that these mAbs stained approximately 1% of peripheral blood T cells, and further showed that the mAbs could stimulate proliferation of these T cells. We then characterized the mAbs by amplifying TCR cDNA derived from mAb-stimulated cells and sequencing the beta chain. All clones sequenced used the V beta 7.1 chain, proving conclusively that the mAbs generated were specific for V beta 7.1 subfamily. This method generates mAbs to human TCR V beta proteins efficiently and might allow production of a complete panel of mAbs directed against human TCR V beta proteins. PMID- 8248132 TI - Ligation of major histocompatibility complex class II molecules mediates apoptotic cell death in resting B lymphocytes. AB - Class II major histocompatibility complex-encoded molecules expressed on the surface of primed B lymphocytes function as restriction elements for presentation of antigen to T lymphocytes, an interaction that ultimately leads to activation and differentiation of both cell types. The engagement of class II on a resting B cell, on the other hand, inhibits subsequent B-cell growth and activation. Our studies show that treatment of resting B lymphocytes with anti-class II antibodies, or with other agents (dibutyryl cAMP or isoproterenol) that increase intracellular levels of cAMP, results in the apoptotic death of most or all of the resting B cells. Conversely, treating cells with immobilized anti immunoglobulin and interleukin 4, conditions known to prime cells, protects them from class II-mediated death and specifically from increases in nucleosomal fragments characteristic of apoptotic death. Freshly ex vivo activated B cells likewise are refractory to class II-mediated apoptosis. Treating B cells with anti-class II reagents causes an elevation of cAMP in resting, but not in activated, B cells. These results suggest that apoptotic death is a mechanism of prevention of nonspecific B-cell activation in the event that T-cell receptor and/or CD4 ligation of major histocompatibility complex class II occurs in the absence of antigen. PMID- 8248133 TI - Relationship between intrinsic connections and functional architecture revealed by optical imaging and in vivo targeted biocytin injections in primate striate cortex. AB - In primate primary visual cortex, neurons sharing similar response properties are clustered together forming functional domains that appear as a mosaic of patches or bands, often traversing the entire cortical depth from the pia to the white matter. Similarly, each cortical site connects laterally through an extensive network of intrinsic projections that are organized in multiple clusters (patches) and reach distances of up to a few millimeters. The relationship between the functional domains and these laterally connected patches has remained a controversial issue despite intensive research efforts. To investigate this relationship, we obtained high-resolution functional maps of the cortical architecture by in vivo optical imaging. Subsequently, extracellular injections of the sensitive anterograde tracer biocytin were targeted into selected functional domains. Within the ocular dominance system, we found that long-range intrinsic connections tended to link the monocular regions of same-eye ocular dominance columns. Furthermore, we discovered that binocular domains formed a separate set of connections in area V1; binocular regions were selectively connected among themselves but were not connected to strictly monocular regions, suggesting that they constitute a distinct columnar system. In the other subsystem subserving orientation preference, patches of intrinsic connections tended to link domains sharing similar orientation preferences. Analyses of the precision of these connections indicated that in both functional subsystems, < 15% of the connections were between domains having orthogonal response properties. However, their selectivity was limited; approximately 30% +/- 10% of the interconnected patches contained neurons exhibiting orientation tuning that differed from those found at the injection sites by at least 45 degrees. At short range (up to 400 microns from the injection site), this casual trend seemed markedly accentuated; the local, synaptic-rich axonal and dendritic arbors crossed freely through columns of diverse functional properties. These complex sets of connections can endow cortical neurons with a rich diversity of response properties and broad tuning. PMID- 8248134 TI - Cloning of the human homolog of the CDC34 cell cycle gene by complementation in yeast. AB - In a screen designed to isolate human cDNAs that complement a yeast G2 phase checkpoint mutation (mec1), we isolated a cDNA homologous to the Saccharomyces cerevisiae CDC34 gene. The human CDC34 cDNA can functionally substitute for the yeast CDC34 gene and represents a mammalian homolog of the group of yeast genes required for the late G1-->S phase transition. The human CDC34 gene is expressed in multiple cell lines as a unique species and Southern blot analysis reveals evidence for a single gene that is highly conserved in higher eukaryotes. The human gene is located on the far telomeric region of 19p13.3 in a location that defines a region of homology between human chromosome 19p and mouse chromosome 11. PMID- 8248135 TI - Topography of the histone octamer surface: repeating structural motifs utilized in the docking of nucleosomal DNA. AB - The histone octamer core of the nucleosome is a protein superhelix of four spirally arrayed histone dimers. The cylindrical face of this superhelix is marked by intradimer and interdimer pseudodyad axes, which derive from the nature of the histone fold. The histone fold appears as the result of a tandem, parallel duplication of the "helix-strand-helix" motif. This motif, by its occurrence in the four dimers, gives rise to repetitive structural elements--i.e., the "parallel beta bridges" and the "paired ends of helix I" motifs. A preponderance of positive charges on the surface of the octamer appears as a left-handed spiral situated at the expected path of the DNA. We have matched a subset of DNA pseudodyads with the octamer pseudodyads and thus have built a model of the nucleosome. In it, the two DNA strands coincide with the path of the histone positive charges, and the central 12 turns of the double helix contact the surface of the octamer at the repetitive structural motifs. The properties of these complementary contacts appear to explain the preference of histones for double-helical DNA and to suggest a possible basis for allosteric regulation of nucleosome function. PMID- 8248136 TI - Preferential repair of ionizing radiation-induced damage in the transcribed strand of an active human gene is defective in Cockayne syndrome. AB - Cells from patients with Cockayne syndrome (CS), which are sensitive to killing by UV although overall damage removal appears normal, are specifically defective in repair of UV damage in actively transcribed genes. Because several CS strains display cross-sensitivity to killing by ionizing radiation, we examined whether ionizing radiation-induced damage in active genes is preferentially repaired by normal cells and whether the radiosensitivity of CS cells can be explained by a defect in this process. We found that ionizing radiation-induced damage was repaired more rapidly in the transcriptionally active metallothionein IIA (MTIIA) gene than in the inactive MTIIB gene or in the genome overall in normal cells as a result of faster repair on the transcribed strand of MTIIA. Cells of the radiosensitive CS strain CS1AN are completely defective in this strand-selective repair of ionizing radiation-induced damage, although their overall repair rate appears normal. CS3BE cells, which are intermediate in radiosensitivity, do exhibit more rapid repair of the transcribed strand but at a reduced rate compared to normal cells. Xeroderma pigmentosum complementation group A cells, which are hypersensitive to UV light because of a defect in the nucleotide excision repair pathway but do not show increased sensitivity to ionizing radiation, preferentially repair ionizing radiation-induced damage on the transcribed strand of MTIIA. Thus, the ability to rapidly repair ionizing radiation-induced damage in actively transcribing genes correlates with cell survival. Our results extend the generality of preferential repair in active genes to include damage other than bulky lesions. PMID- 8248137 TI - Electroporation of mammalian skin: a mechanism to enhance transdermal drug delivery. AB - Mammalian skin owes its remarkable barrier function to its outermost and dead layer, the stratum corneum. Transdermal transport through this region occurs predominantly through intercellular lipids, organized largely in bilayers. Electroporation is the creation of aqueous pores in lipid bilayers by the application of a short (microseconds to milliseconds) electric pulse. Our measurements suggest that electroporation occurs in the intercellular lipid bilayers of the stratum corneum by a mechanism involving transient structural changes. Flux increases up to 4 orders of magnitude were observed with human skin in vitro for three polar molecules having charges between -1 and -4 and molecular weights up to slightly more than 1000. Similar flux increases were observed in vivo with animal skin. These results may have significance for drug delivery and other medical applications. PMID- 8248138 TI - Identification of barriers to rotation of DNA segments in yeast from the topology of DNA rings excised by an inducible site-specific recombinase. AB - Controlled excision of DNA segments to yield intracellular DNA rings of well defined sequences was utilized to study the determinants of transcriptional supercoiling of closed circular DNA in the yeast Saccharomyces cerevisiae. In delta top1 top2ts strains of S. cerevisiae expressing Escherichia coli DNA topoisomerase I, accumulation of positive supercoils in intracellular DNA normally occurs upon thermal inactivation of DNA topoisomerase II because of the simultaneous generation of positively and negatively supercoiled domains by transcription and the preferential relaxation of the latter by the bacterial enzyme. Positive supercoil accumulation in DNA rings is shown to depend on the presence of specific sequence elements; one likely cause of this dependence is that the persistence of oppositely supercoiled domains in an intracellular DNA ring requires the presence of barriers to rotation of the DNA segments connecting the domains. Analysis of the S. cerevisiae 2-microns plasmid partition system by this approach suggests that the plasmid-encoded REP1 and REP2 proteins are involved in forming such a barrier in DNA containing the REP3 sequence. PMID- 8248139 TI - De novo synthesis of budding yeast DNA polymerase alpha and POL1 transcription at the G1/S boundary are not required for entrance into S phase. AB - The POL1 gene, encoding DNA polymerase alpha (pol alpha) in Saccharomyces cerevisiae, is transiently transcribed during the cell cycle at the G1/S phase boundary. Here we show that yeast pol alpha is present at every stage of the cell cycle, and its level only slightly increases following the peak of POL1 transcription. POL1 mRNA synthesis driven by a GAL1 promoter can be completely abolished without affecting the growth rate of logarithmically growing yeast cultures for several cell divisions, although the amount of the pol alpha polypeptide drops below the physiological level. Moreover, alpha-factor-arrested cells can enter S phase and divide synchronously even if POL1 transcription is abolished. These results indicate that the level of yeast pol alpha is not rate limiting and de novo synthesis of the enzyme is not required for entrance into S phase. PMID- 8248140 TI - Automatic enumeration and characterization of heterogeneous clonal progression in cell transformation. AB - Most human tumors are clonal in origin, although the cells may be diverse in their properties. Since the tumors evolve through progressive stages over decades of time, it is possible that the conditions that induced the tumor transform many cells, but that selective overgrowth of the fastest growing lead to a clonal population of identified tumor cells. We studied the progression of neoplastic transformation in clones from a population in which about 10% of the cells formed well-defined transformed foci. A few of the clones produced many large foci, but most of the clones produced no foci or only one focus. Maintenance of the nonproducing and low-producing clones under the growth constraint of confluence and low serum concentration, which promotes transformation, led to the production of large numbers of small foci by all of them. Visual inspection revealed considerable heterogeneity in size and density among the foci from each clone, and this was quantitated by computer scanning. Subclonal analysis of focus formation was done on one of the clones after it had undergone further growth constraint to promote transformation. As in the original cloning, some of the subclones produced many large foci, but most produced none. Another round of growth constraint was imposed on the nonproducing subclones, which then became producers of many small foci varying in size and density. The results indicate that most if not all cells in the population respond to growth constraint by undergoing transformation. Though there is wide variation in the degree of transformation, the results are consistent with the view that an entire field of cells exposed to carcinogenic conditions in an animal undergoes some progression toward neoplasia but that heterogeneity of the response followed by selective neoplastic growth may lead to a clonal origin of the clinically detected tumor. PMID- 8248141 TI - Specific UV-induced mutation spectrum in the p53 gene of skin tumors from DNA repair-deficient xeroderma pigmentosum patients. AB - The UV component of sunlight is the major carcinogen involved in the etiology of skin cancers. We have studied the rare, hereditary syndrome xeroderma pigmentosum (XP), which is characterized by a very high incidence of cutaneous tumors on exposed skin at an early age, probably due to a deficiency in excision repair of UV-induced lesions. It is interesting to determine the UV mutation spectrum in XP skin tumors in order to correlate the absence of repair of specific DNA lesions and the initiation of skin tumors. The p53 gene is frequently mutated in human cancers and represents a good target for studying mutation spectra since there are > 100 potential sites for phenotypic mutations. Using reverse transcription PCR and single-strand conformation polymorphism to analyze > 40 XP skin tumors (mainly basal and squamous cell carcinomas), we have found that 40% (17 out of 43) contained at least one point mutation on the p53 gene. All the mutations were located at dipyrimidine sites, essentially at CC sequences, which are hot spots for UV-induced DNA lesions. Sixty-one percent of these mutations were tandem CC- >TT mutations considered to be unique to UV-induced lesions; these mutations are not observed in internal human tumors. All the mutations, except two, must be due to translesion synthesis of unrepaired dipyrimidine lesions left on the nontranscribed strand. These results show the existence of preferential repair of UV lesions [either pyrimidine dimers or pyrimidine-pyrimidone (6-4) photoproducts] on the transcribed strand in human tissues. PMID- 8248142 TI - RNA sequence of astrovirus: distinctive genomic organization and a putative retrovirus-like ribosomal frameshifting signal that directs the viral replicase synthesis. AB - The genomic RNA of human astrovirus was sequenced and found to contain 6797 nt organized into three open reading frames (1a, 1b, and 2). A potential ribosomal frameshift site identified in the overlap region of open reading frames 1a and 1b consists of a "shifty" heptanucleotide and an RNA stem-loop structure that closely resemble those at the gag-pro junction of some retroviruses. This translation frame-shift may result in the suppression of in-frame amber termination at the end of open reading frame 1a and the synthesis of a nonstructural, fusion polyprotein that contains the putative protease and RNA dependent RNA polymerase. Comparative sequence analysis indicated that the protease and polymerase of astrovirus are only distantly related to the respective enzymes of other positive-strand RNA viruses. The astrovirus polyprotein lacks the RNA helicase domain typical of other positive-strand RNA viruses of similar genome size. The genomic organization and expression strategy of astrovirus, with the protease and the polymerase brought together by predicted frameshift, most closely resembled those of plant leuteoviruses. Specific features of the sequence and genomic organization support the classification of astroviruses as an additional family of positive-strand RNA viruses, designated Astroviridae. PMID- 8248143 TI - Salmonella induces the formation of filamentous structures containing lysosomal membrane glycoproteins in epithelial cells. AB - Salmonella species invade and replicate within epithelial cells in membrane-bound vacuoles. In this report we show that upon infection of HeLa epithelial cells, Salmonella typhimurium residues in vacuoles that contain lysosomal membrane glycoproteins (lgps). Four to six hours after invasion, intracellular bacteria induce the formation of stable filamentous structures containing lgps that are connected to the bacteria-containing vacuoles. Formation of these lgp-rich structures requires viable intracellular bacteria and is blocked by inhibitors of vacuolar acidification. These structures are not present in uninfected cells or in cells infected with another invasive bacteria, Yersinia enterocolitica. Tracers added to the extracellular medium are not delivered to the Salmonella induced filaments, suggesting that these structures are different from previously described tubular lysosomes. Initiation of intracellular bacterial replication correlates with formation of these lgp-containing filaments. Certain avirulent Salmonella mutants that are defective for intracellular replication fail to induce formation of these structures. These observations suggest that Salmonella induced filaments containing lgps are linked to intracellular bacterial replication. PMID- 8248144 TI - Mechanistic aspects of genome-wide demethylation in the preimplantation mouse embryo. AB - Gene-specific methylation patterns in mammals play a role in a variety of biological processes in the embryo and adult tissues. These patterns are established during embryo development by a process that involves genome-wide demethylation in the morula and de novo methylation in the pregastrula. To elucidate the mechanism of demethylation in the early mouse embryo, we have injected mouse zygotes with gene sequences that were methylated in vitro by Hpa II methylase and analyzed the methylation status of specific sites in blastocyst DNA. Because it had been propagated in Escherichia coli, the DNA used for these injections was also methylated at adenine residues in GATC sites. This allowed us to eliminate fully methylated, unintegrated DNA by Dpn I digestion and fully unmethylated, integrated DNA that underwent several rounds of replication by Mbo I digestion. The integrated, originally injected DNA strands were in a hemimethylated state and survived this treatment. The methylation status of Hpa II sites in these molecules was analyzed by Hpa II digestion of the genomic DNA isolated from blastocysts, followed by PCR amplification using appropriate primers. The results demonstrate that demethylation is achieved by an active mechanism and that specific sites in imprinted genes escape demethylation, maintaining a methylated state throughout preimplantation development. PMID- 8248145 TI - Ectopic expression of wild-type or a dominant-negative mutant of transcription factor NTF-1 disrupts normal Drosophila development. AB - The Drosophila melanogaster tissue-specific transcription factor NTF-1 was originally identified in vitro as a protein that could bind to and activate transcription from the Dopa decarboxylase (Ddc) gene. A structure-function analysis of NTF-1 led to the identification of a discrete amino-terminal activation domain. Here, we report that an NTF-1 mutant lacking the activation domain acts as a trans-dominant inhibitor of NTF-1 activation in tissue culture cells by forming inactive heterodimers with the full-length protein. Ectopically expressing this dominant-negative protein or the full-length protein in developing Drosophila embryos leads to dire developmental consequences. Overexpressing the trans-dominant NTF-1 leads to lethality, while overexpressing full-length NTF-1 results in both lethality and morphogenetic defects. Our results suggest that both the activity and the regulation of NTF-1 are critical for viability and proper development of the fly. PMID- 8248146 TI - Comparative spectroscopic, calorimetric, and computational studies of nucleic acid complexes with 2',5"-versus 3',5"-phosphodiester linkages. AB - We have used a combination of spectroscopic, calorimetric, and computational techniques to characterize the properties of nucleic acid complexes with 2',5''- and 3',5''-phosphodiester linkages. Specifically, we have compared the properties of complexes formed by the association of 3',5'' single-stranded 16-mers of adenylic acid (A16) and thymidylic acid (T16) with the complexes formed by the corresponding single-stranded 16-mers with 2',5''-phosphodiester linkages (A*16 and T*16). Our results reveal the following differential features: (i) the 3',5'' strands form either a duplex or a triplex, depending on the sodium ion concentration, whereas the 2',5'' strands form either a triplex or no complex at all; (ii) the 2',5'' and 3',5'' triplexes exhibit significantly different CD spectra, suggesting that the two triplex states are conformationally nonequivalent; (iii) the 2',5'' triplex has a lower charge density than the 3',5'' triplex; (iv) the thermal stability of the 3',5'' triplex, as expected, is concentration dependent, whereas the thermal stability of the 2',5'' triplex is concentration independent; (v) relative to their component single strands, the 2',5'' triplex is thermodynamically much less stable than the 3',5'' triplex, despite being thermally more stable; (vi) the reduced thermodynamic stability of the 2',5'' triplex relative to the 3',5'' triplex is overwhelmingly enthalpic in origin. In the aggregate, our results reveal and characterize significant differences in the properties of complexes formed by the association of strands with identical base sequences but different phosphodiester linkages. We describe a structural model that is consistent with many of the differential properties observed. We also speculate on how these differential properties may have provided an evolutionary advantage for 3',5'' linkages and how the properties of 2',5'' complexes might be exploited in antisense strategies. PMID- 8248147 TI - Expression of the human beta-amyloid precursor protein gene from a yeast artificial chromosome in transgenic mice. AB - One hallmark of Alzheimer disease is the formation in the brain of amyloid plaques containing a small peptide derived from the beta-amyloid precursor protein (APP). The APP gene exhibits a complex pattern of expression in peripheral tissues and in the brain. The entire human APP gene was introduced into embryonic stem (ES) cells by co-lipofection of a 650-kb yeast artificial chromosome (YAC). Three ES lines containing an essentially intact YAC were isolated, and expression of human APP mRNAs at levels comparable to those of endogenous mouse APP transcripts was obtained. A transgenic mouse line was established by germ-line transmission of the APP YAC. RNase protection analysis of human APP mRNAs demonstrated appropriate splicing of the primary APP transcript in ES cells and in the brain of a transgenic animal. These mice may be useful for elucidating the function of the various APP isoforms in vivo. PMID- 8248148 TI - A second hepatitis C virus-encoded proteinase. AB - Host and viral proteinases are believed to be required for the production of at least nine hepatitis C virus (HCV)-specific polyprotein cleavage products. Although several cleavages appear to be catalyzed by host signal peptidase or the HCV NS3 serine proteinase, the enzyme responsible for cleavage at the 2/3 site has not been identified. In this report, we have defined the 2/3 cleavage site and obtained evidence which suggests that this cleavage is mediated by a second HCV-encoded proteinase, located between aa 827 and 1207. This region encompasses the C-terminal portion of the 23-kDa NS2 protein, the 2/3 cleavage site, and the serine proteinase domain of NS3. Efficient processing at the 2/3 site was observed in mammalian cells, Escherichia coli, and in plant or animal cell-free translation systems in the absence of microsomal membranes. Cleavage at the 2/3 site was abolished by alanine substitutions for NS2 residues His-952 or Cys-993 but was unaffected by several other substitution mutations, including those that inactivate NS3 serine proteinase function. Mutations abolishing cleavage at the 2/3 site did not block cleavage at other sites in the HCV polyprotein. Cotransfection experiments indicate that the 2/3 site can be cleaved in trans, which should facilitate purification and further characterization of this enzyme. PMID- 8248149 TI - The lysosomal proenzyme receptor that binds procathepsin L to microsomal membranes at pH 5 is a 43-kDa integral membrane protein. AB - Two lysosomal proenzymes, procathepsins L and D, bind to mouse fibroblast microsomal membranes at acidic pH. This membrane association is independent of the mannose-6-phosphate receptors and requires the presence of the N-terminal propeptides of the enzymes. We have identified the protein that specifically binds procathepsin L at pH 5. A 43-kDa membrane protein coimmunoprecipitated with procathepsin L at pH 5 but not at pH 7 when cells were denatured with detergents. Similarly, a 43-kDa integral membrane protein bound procathepsin L in three kinds of ligand blots at pH 5 but not at pH 7. A synthetic peptide containing the 24 N terminal residues of mouse procathepsin L blocked the binding of procathepsin L to this integral membrane protein on ligand blots. These results indicate that the 43-kDa integral membrane protein is a lysosomal proenzyme receptor that specifically binds the procathepsin L activation peptide at acidic pH. PMID- 8248150 TI - Genetic analysis of homomeric interactions of human immunodeficiency virus type 1 integrase using the yeast two-hybrid system. AB - The retroviral integrase protein (IN) is responsible for catalyzing a concerted integration reaction in which the two termini of linear viral DNA are joined to host DNA. To probe the potential for IN to form protein multimers, we used the yeast two-hybrid system. The coexpression of a GAL4 DNA binding domain-IN fusion and a GAL4 activation domain-IN fusion together resulted in the successful activation of a GAL4-responsive LacZ reporter gene. The system was used to examine a variety of IN deletion mutants. The results suggest that the central region of the protein is necessary for multimerization and that the N-terminal zinc finger region is not important. PMID- 8248151 TI - Multiple loci for synapse protein SNAP-25 in the tetraploid goldfish. AB - The common goldfish Carassius auratus is tetraploid and has 100 chromosomes. We describe here goldfish cDNA clones for SNAP-25, a 200-amino-acid synaptosome associated protein that has remained highly conserved during evolution. SNAP-25 occurs as a single-copy gene in mouse, chicken, and Drosophila melanogaster. Sequences of six distinct goldfish cDNA clones and Southern hybridizations show that the goldfish has three, or possibly four, SNAP-25 loci rather than two as expected. A gene duplication early in actinopterygian fish evolution gave rise to the loci SnapA and SnapB. The proteins SNAP-A and SNAP-B are 94% and 91% identical to the mouse protein but are only 91% identical to each other. SNAP-B has a larger number of unique amino acid replacements than SNAP-A and also has more dramatic replacements. The tetraploidization resulted in two SnapB loci whose divergence from each other is consistent with a tetraploidization event 15 20 million years ago. The presence of duplicate SnapA loci has not yet been possible to confirm, possibly because they are still very similar to each other. Two of the SnapA cDNA clones and one SnapB cDNA clone have frameshift mutations. As these aberrant alleles otherwise display high sequence identity to the functional alleles, they probably became nonfunctional recently. The findings of allelic variability and aberrant alleles emphasize the importance of characterizing multiple DNA clones in tetraploid species. PMID- 8248153 TI - Conservation of allelic richness in wild crop relatives is aided by assessment of genetic markers. AB - Wild crop relatives are an important source of genetic variation for improving domesticated species. Given limited resources, methods for maximizing the genetic diversity of collections of wild relatives are needed to help spread protection over a larger number of populations and species. Simulations were conducted to investigate the optimal strategy of sampling materials from populations of wild relatives, with the objective of maximizing the number of alleles (allelic richness) in collections of fixed size. Two methods, based on assessing populations for variation at marker loci (e.g., allozymes, restriction fragment length polymorphisms), were developed and compared with several methods that are not dependent on markers. Marker-assisted methods yielded higher overall allelic richness in the simulated collections, and they were particularly effective in conserving geographically localized alleles, the class of alleles that is most subject to loss. PMID- 8248152 TI - Isolation of a shark immunoglobulin light chain cDNA clone encoding a protein resembling mammalian kappa light chains: implications for the evolution of light chains. AB - The time of emergence of immunoglobulin kappa and lambda light (L) chains in evolution is unknown. An L chain cDNA clone was isolated from the nurse shark (Ginglymostoma cirratum), a cartilaginous fish, whose predicted variable (V) region amino acid sequence has up to 60% sequence identity to mammalian V kappa domains. Genomic analyses suggest a cluster-type gene organization for this L chain locus, similar to the shark lambda-like immunoglobulin L chain loci rather than mammalian kappa loci. We propose that divergence of the ancestral L chain into isotypes likely occurred before the emergence of elasmobranchs 400-450 million years ago. Similarities in gene organization between the two isotypes in sharks may reflect the gene organization utilized by the ancestral L chain. PMID- 8248154 TI - Chromatin structure and regulation of the eukaryotic regulatory gene GAL80. AB - The chromatin structure around the 5' end of the yeast regulatory gene GAL80 has been determined. The chromatin organization is very similar to that on the 5' regions of the GAL1-10 structural genes: a constitutive hypersensitive region containing the upstream activating sequence (UAS) element, and nucleosomes around this hypersensitive region. The downstream nucleosome, which is a positioned nucleosome, covers the TATA and transcription start sites. The nucleosome upstream of the hypersensitive region undergoes significant change when cells are grown in galactose, where GAL80 gene expression is induced to maximal levels. The change may be related to the induction process. GAL4 protein binds strongly to the GAL80 UAS in galactose-grown cells, less strongly in glycerol-grown cells, and not at all in glucose-grown cells. These data and published gene expression data are used to develop a model for the regulation of the GAL80 regulatory gene. PMID- 8248155 TI - Expression and biochemical properties of a protein serine/threonine phosphatase encoded by bacteriophage lambda. AB - The predicted amino acid sequence encoded by the open reading frame 221 (orf221) of bacteriophage lambda exhibited a high degree of similarity to the catalytic subunits of a variety of protein serine/threonine phosphatases belonging to PP1, PP2A, and PP2B groups. Cloning and expression of the orf221 gene in Escherichia coli provided direct evidence that the gene codes for a protein serine/threonine phosphatase. The single-subunit recombinant enzyme was purified in soluble form and shown to possess a unique repertoire of biochemical properties--e.g., an absolute requirement for Mn2+, resistance to okadaic acid, inhibitors 1 and 2, and ability to dephosphorylate casein, adenovirus E1A proteins, and the alpha subunit of phosphorylase kinase. No phosphotyrosine phosphatase activity was observed. Mutational and biochemical analyses identified the conserved residues 73-77 and Cys138 to be important for activity. The name PP-lambda is proposed for this unusual prokaryotic enzyme. PMID- 8248156 TI - Strand-invasion of duplex DNA by peptide nucleic acid oligomers. AB - Polyamide oligomers, termed peptide nucleic acids (PNAs), bind with high affinity to both DNA and RNA and offer both antisense and antigene approaches for regulating gene expression. When a PNA binds to a complementary sequence in a double-stranded DNA, one strand of the duplex is displaced, and a stable D-loop is formed. Unlike oligodeoxynucleotides for which binding polarity is determined by the deoxyribose sugar, the unrestrained polyamide backbone of the PNA could permit binding to a DNA target in an orientation-independent manner. We now provide evidence that PNAs can, in fact, bind to their complementary sequence in DNA independent of the DNA-strand polarity--that is, a PNA binds to DNA in both "parallel" and "antiparallel" fashion. With a mixed-sequence 15-mer PNA, kinetic studies of PNA.DNA interactions revealed that D-loop formation was rapid and the complex was stable for several hours. However, when measured either by gel mobility-shift analysis or RNA polymerase II-elongation termination, D-loop formation was salt dependent, but PNA-strand dissociation was not salt dependent. We observed that D-loop-containing DNA fragments had anomalous gel mobilities that varied as a function of the position of the D-loop relative to the DNA termini. On the basis of permutation analysis, the decreased mobility of the PNA.DNA complex was attributed to a bend in the DNA at or near the D-loop. PMID- 8248157 TI - Genetic and linguistic differentiation in the Americas. AB - The relationship between linguistic differentiation and evolutionary affinities was evaluated in three tribes of the Pacific Northwest. Two tribes (Nuu-Chah Nulth and Bella Coola) speak Amerind languages, while the language of the third (Haida) belongs to a different linguistic phylum--Na-Dene. Construction of a molecular phylogeny gave no evidence of clustering by linguistic affiliation, suggesting a relatively recent ancestry of these linguistically divergent populations. When the evolutionary affinities of the tribes were evaluated in terms of mitochondrial sequence diversity, the Na-Dene-speaking Haida had a reduced amount of diversity compared to the two Amerind tribes and thus appear to be a biologically younger population. Further, since the sequence diversity between the two Amerind-speaking tribes is comparable to the diversity between the Amerind tribes and the Na-Dene Haida, the evolutionary divergence within the Amerind linguistic phylum may be as great as the evolutionary divergence between the Amerind and Na-Dene phyla. Hence, in the New World, rates of linguistic differentiation appear to be markedly faster than rates of biological differentiation, with little congruence between linguistic hierarchy and the pattern of evolutionary relationships. PMID- 8248158 TI - Social control of primary sex differentiation in the Midas cichlid. AB - Sexual differentiation in teleost fishes is characteristically labile. The most dramatic form of sexual lability is postmaturational sex change, which is common among teleosts although rare or absent in other vertebrate taxa. In many cases this process is regulated by social cues, particularly dominance interactions. Here we show that in the Midas cichlid, Cichlasoma citrinellum, these same sorts of social interactions affect much earlier stages of sexual differentiation. In this species, males are larger than females. By manipulating relative size in juveniles, we show that this sex-based size difference does not arise from endogenous factors associated with sex. Rather, sex is determined by relative size as a juvenile. We argue that this mode of sex determination, which may be common among teleosts, is a heterochronic variant of postmaturational sex change, one in which some individuals are deflected from a default female trajectory before maturation, as a result of social signals. The size-advantage model, which specifies the optimal size for sex change in hermaphroditic species, can be extended to account for the decision whether to mature as a male or a female in the Midas cichlid. PMID- 8248159 TI - An examination of the generation-time effect on molecular evolution. AB - By using DNA sequences of 17 mammalian genes, the generation-time effect is estimated separately for synonymous substitutions and nonsynonymous substitutions. Star phylogenies composed of rodentia, artiodactyla, and primates are examined. The generation-time effect is found to be more conspicuous for synonymous substitutions than for non-synonymous substitutions, by using the methods of (i) Nei and Gojobori, (ii) Li, and (iii) Ina. The proportion of accepted amino acid substitutions in evolution is estimated to be about twice as large in the primate lineage as in the rodent lineage. This result is in accord with the nearly neutral theory of molecular evolution. PMID- 8248160 TI - Comparison of somatic mutation in a transgenic versus host locus. AB - Somatic mutations can now be quantified in almost any cell type in mice carrying bacterial genes in a lambda phage shuttle vector. Mutations induced in vivo are detectable ex vivo, after packaging host-cell DNA into phage that are grown on suitable bacteria. However, the transgenic DNA differs from many host loci in several ways: it (i) is prokaryotic DNA, (ii) is present in multiple tandem copies, and (iii) is heavily methylated and probably not expressed. Thus, mutation of a transgene may not be a suitable model of the host loci, which are eukaryotic, unique, and expressed. To test the relevance of the transgene mutation model, the frequencies of the bacterial lacI+ to lacI- mutations induced in half of the small intestine were compared with the frequencies of the host Dlb 1b to Dlb-1a mutations induced in the other half. The loci responded similarly to ethyl nitrosourea (ENU) with respect to the animal's age and sex, sex of the parent transmitting the transgene, and expression time. ENU dose-response curves were similar. Furthermore, no difference was found at the Dlb-1 locus between transgenic and nontransgenic siblings. In contrast, x-rays induced few lacI mutations but many Dlb-1 mutations. Probably few large deletions are detectable at lacI, but many are detectable at Dlb-1. If so, an important class of mutation is not readily detected in these transgenic mice. With this exception, the transgene and host gene responded similarly in this somewhat limited trial, as is necessary if the transgenic mice are to be a useful model. PMID- 8248161 TI - cDNA cloning, characterization, and tissue-specific expression of human xanthine dehydrogenase/xanthine oxidase. AB - We isolated cDNAs encoding xanthine dehydrogenase (XD; xanthine:NAD+ oxidoreductase, EC 1.1.1.204) from a human liver cDNA library. The complete nucleotide sequence of human XD was determined; the deduced amino acid sequence encoded a protein of 1336 amino acid residues of M(r) 147,782. Human XD possessed many of the signature sequences typical of XDs from flies and rodents, including an unusual cysteine distribution, a potential 2Fe/2S binding site, and a putative molybdopterin cofactor binding domain. Analysis of potential NAD binding sites suggested a simple hypothesis for the conversion of human XD into the oxygen metabolite forming xanthine oxidase (XO; xanthine:oxygen oxidoreductase, EC 1.1.3.22). Using a human XD complementary RNA hybridization probe, we found a 5100-base RNA in human liver by RNA blot-hybridization analysis. This RNA exhibited tissue-specific distribution that may be pertinent to XD- and XO mediated oxygen radical injury in ischemia/reperfusion and inflammation. A second 4500-base RNA was detected in some tissues and may arise through differential transcription termination. PMID- 8248162 TI - Inhibition of vascular endothelial cell growth factor activity by an endogenously encoded soluble receptor. AB - Vascular endothelial cell growth factor, a mitogen selective for vascular endothelial cells in vitro that promotes angiogenesis in vivo, functions through distinct membrane-spanning tyrosine kinase receptors. The cDNA encoding a soluble truncated form of one such receptor, fms-like tyrosine kinase receptor, has been cloned from a human vascular endothelial cell library. The mRNA coding region distinctive to this cDNA has been confirmed to be present in vascular endothelial cells. Soluble fms-like tyrosine kinase receptor mRNA, generated by alternative splicing of the same pre-mRNA used to produce the full-length membrane-spanning receptor, encodes the six N-terminal immunoglobulin-like extracellular ligand binding domains but does not encode the last such domain, transmembrane-spanning region, and intracellular tyrosine kinase domains. The recombinant soluble human receptor binds vascular endothelial cell growth factor with high affinity and inhibits its mitogenic activity for vascular endothelial cells; thus this soluble receptor could act as an efficient specific antagonist of vascular endothelial cell growth factor in vivo. PMID- 8248163 TI - RNA fingerprinting using arbitrarily primed PCR identifies differentially regulated RNAs in mink lung (Mv1Lu) cells growth arrested by transforming growth factor beta 1. AB - RNA fingerprinting using arbitrarily primed PCR (RAP) samples an RNA population and allows the detection of differentially expressed genes between two or more populations. This method was applied to mink lung epithelial cells, which respond to treatment with transforming growth factor beta (TGF-beta) by undergoing cell cycle arrest at or near the G1/S-phase boundary. The steady-state abundances of approximately 200 RNAs were surveyed, a few of which displayed differential regulation in response to TGF-beta 1. Three products were isolated, cloned, and sequenced. One differentially regulated RNA corresponded to cyclin A, a gene known to be required for the progression of mammalian fibroblasts through S phase. Northern blot analysis confirmed that the cyclin A mRNA steady-state abundance decreased dramatically in response to a 24-hr TGF-beta 1 treatment and also in response to cell cycle arrest caused by contact inhibition. A second RAP product corresponded to a previously unknown 7.5-kb mRNA, the level of which decreased dramatically in response to TGF-beta 1 treatment. Unlike the cyclin A mRNA, the abundance of this transcript did not decrease in response to growth arrest induced by contact inhibition. A third RAP product corresponded to the mRNA for osteonectin, an extracellular matrix protein. The abundance of this mRNA increased at least 2-fold during TGF-beta 1 treatment. This observation is consistent with other reports of increases in extracellular matrix proteins during TGF-beta treatment. RAP should be able to identify many of the genes that change in steady-state expression during the cell cycle. PMID- 8248164 TI - Cellular epigenetics: effects of passage history on competence of cells for "spontaneous" transformation. AB - "Spontaneous" neoplastic transformation of cells in culture has been shown to be an adaptive response to moderate physiological constraints such as contact inhibition and lowered concentrations of serum. These are the same constraints that promote normal differentiation of a variety of cell types. Because both normal and neoplastic development represent enduring changes in response to constraint, spontaneous transformation can be considered a form of epigenesis. This phenomenon permits the use of established cell lines in developing general epigenetic principles with all of the advantages of manipulation, efficient cloning, and quantification afforded by such systems. Being environmentally sensitive, however, the responsive capacity of cell lines is subject to fluctuation and long-term modification. I therefore set out to define the conditions used in maintaining cells that influence their competence to undergo transformation with the aim of controlling that competence. It proved to be sensitive to both the population density and frequency of three different repetitive passage regimens. Both the saturation density of the cells and their capacity to produce transformed foci varied initially in different ways within the same regimen in two consecutive passage series starting from the same frozen stock, but both parameters eventually settled down to values characteristic of each of the three regimens. As a result, three sublines were developed of high, intermediate, and low competence for transformation. Each of the sublines has special advantages for studying different aspects of epigenetic change. In the process of developing the sublines, a number of observations were made that reinforce the epigenetic nature of neoplastic transformation and are consistent with the concept of progressive state selection as the basis of the change. That concept can be considered a first step in the formal analysis of epigenesis. PMID- 8248165 TI - Resolution of the HLA-DRB6 puzzle: a case of grafting a de novo-generated exon on an existing gene. AB - HLA-DRB6, one of the human major histocompatibility complex genes, lacks exon 1, which normally codes for the leader and the first four amino acid residues of the mature protein. Because it also lacks the HLA promoter, it was surprising to find that the gene is transcribed at a low level in a chimpanzee B-lymphoblastoma cell line, in which the DRB6 homolog is truncated as in humans. The study designed to resolve the paradox has revealed that a retrovirus related to the mouse mammary tumor viruses was inserted into intron 1 of DRB6 > 23 million years ago. The insertion was either accompanied or followed by the deletion of exon 1 and the promoter region of DRB6. In the 3' long terminal repeat of the retrovirus, however, an open reading frame for a new exon arose, which codes for a sequence of mostly hydrophobic amino acid residues; the sequence could function as a leader for the truncated DRB6 gene. This new exon has a functional donor splice site at its 3' end, which enables it to be spliced in register with DRB6 exon 2. Upstream from the new exon is a promoter enabling transcription of the DRB6 gene. Besides providing an example of a de novo generation of an exon, the study suggests a potential mechanism for generating new genes through the replacement of old exons with newly generated ones. PMID- 8248166 TI - Simple models for reading neuronal population codes. AB - In many neural systems, sensory information is distributed throughout a population of neurons. We study simple neural network models for extracting this information. The inputs to the networks are the stochastic responses of a population of sensory neurons tuned to directional stimuli. The performance of each network model in psychophysical tasks is compared with that of the optimal maximum likelihood procedure. As a model of direction estimation in two dimensions, we consider a linear network that computes a population vector. Its performance depends on the width of the population tuning curves and is maximal for width, which increases with the level of background activity. Although for narrowly tuned neurons the performance of the population vector is significantly inferior to that of maximum likelihood estimation, the difference between the two is small when the tuning is broad. For direction discrimination, we consider two models: a perceptron with fully adaptive weights and a network made by adding an adaptive second layer to the population vector network. We calculate the error rates of these networks after exhaustive training to a particular direction. By testing on the full range of possible directions, the extent of transfer of training to novel stimuli can be calculated. It is found that for threshold linear networks the transfer of perceptual learning is nonmonotonic. Although performance deteriorates away from the training stimulus, it peaks again at an intermediate angle. This nonmonotonicity provides an important psychophysical test of these models. PMID- 8248167 TI - Overexpression of human loricrin in transgenic mice produces a normal phenotype. AB - The cornified cell envelope (CE) of terminally differentiating stratified squamous epithelial cells is a complex multiprotein assembly about 15 nm thick of which loricrin is a major component. We have produced transgenic mice bearing the human loricrin transgene in order to study the role of loricrin in CE assembly, structure, and function. By analyses of RNA and protein, we show that the human transgene is expressed in mouse epithelial tissues in an appropriate developmental manner but at an overall level about twice that of endogenous mouse loricrin. Thus the 20-kbp construct used contains all necessary regulatory elements. By immunogold electron microscopy, all of the expressed protein is incorporated into the CE. That no alternations were noted indicates that overproduction of the loricrin component of the CE does not affect the flexible structure or function of the epithelial tissues. Furthermore, these data imply that loricrin may be the last protein to be deposited onto, and thus lines, the intracellular surface of the CE, where it may be accessible to interact with the subjacent keratin intermediate-filament network. PMID- 8248168 TI - Direct transfer of transforming growth factor beta 1 gene into arteries stimulates fibrocellular hyperplasia. AB - The arterial wall responds to thrombosis or mechanical injury through the induction of specific gene products that increase cellular proliferation and connective tissue formation. These changes result in intimal hyperplasia that is observed in restenosis and the early phases of atherosclerosis. Transforming growth factor beta 1 (TGF-beta 1) is a secreted multi-functional protein that plays an important role in embryonal development and in repair following tissue injury. However, the function of TGF-beta 1 in vascular cell growth in vivo has not been defined. In this report, we have evaluated the role of TGF-beta 1 in the pathophysiology of intimal and medial hyperplasia by gene transfer of an expression plasmid encoding active TGF-beta 1 into porcine arteries. Expression of TGF-beta 1 in normal arteries resulted in substantial extracellular matrix production accompanied by intimal and medial hyperplasia. Increased procollagen, collagen, and proteoglycan synthesis in the neointima was demonstrated by immunohistochemistry relative to control transfected arteries. Expression of TGF beta 1 induced a distinctly different program of gene expression and biologic response from the platelet-derived growth factor B (PDGF B) gene: procollagen synthesis induced by TGF-beta 1 was greater, and cellular proliferation was less prominent. These findings show that TGF-beta 1 differentially modulates extracellular matrix production and cellular proliferation in the arterial wall in vivo and could play a reparative role in the response to arterial injury. PMID- 8248169 TI - Intraarticular expression of biologically active interleukin 1-receptor antagonist protein by ex vivo gene transfer. AB - Gene therapy offers a radical different approach to the treatment of arthritis. Here we have demonstrated that two marker genes (lacZ and neo) and cDNA coding for a potentially therapeutic protein (human interleukin 1-receptor-antagonist protein; IRAP or IL-1ra) can be delivered, by ex vivo techniques, to the synovial lining of joints; intraarticular expression of IRAP inhibited intraarticular responses to interleukin 1. To achieve this, lapine synoviocytes were first transduced in culture by retroviral infection. The genetically modified synovial cells were then transplanted by intraarticular injection into the knee joints of rabbits, where they efficiently colonized the synovium. Assay of joint lavages confirmed the in vivo expression of biologically active human IRAP. With allografted cells, IRAP expression was lost by 12 days after transfer. In contrast, autografted synoviocytes continued to express IRAP for approximately 5 weeks. Knee joints expressing human IRAP were protected from the leukocytosis that otherwise follows the intraarticular injection of recombinant human interleukin 1 beta. Thus, we report the intraarticular expression and activity of a potentially therapeutic protein by gene-transfer technology; these experiments demonstrate the feasibility of treating arthritis and other joint disorders with gene therapy. PMID- 8248170 TI - The precursor region of a protein active in sperm-egg fusion contains a metalloprotease and a disintegrin domain: structural, functional, and evolutionary implications. AB - PH-30, a sperm surface protein involved in sperm-egg fusion, is composed of two subunits, alpha and beta, which are synthesized as precursors and processed, during sperm development, to yield the mature forms. The mature PH-30 alpha/beta complex resembles certain viral fusion proteins in membrane topology and predicted binding and fusion functions. Furthermore, the mature subunits are similar in sequence to each other and to a family of disintegrin domain containing snake venom proteins. We report here the sequences of the PH-30 alpha and beta precursor regions. Their domain organizations are similar to each other and to precursors of snake venom metalloproteases and disintegrins. The alpha precursor region contains, from amino to carboxyl terminus, pro, metalloprotease, and disintegrin domains. The beta precursor region contains pro and metalloprotease domains. Residues diagnostic of a catalytically active metalloprotease are present in the alpha, but not the beta, precursor region. We propose that the active sites of the PH-30 alpha and snake venom metalloproteases are structurally similar to that of astacin. PH-30, acting through its metalloprotease and/or disintegrin domains, could be involved in sperm development as well as sperm-egg binding and fusion. Phylogenetic analysis indicates that PH-30 stems from a multidomain ancestral protein. PMID- 8248171 TI - Characterization of broken DNA molecules associated with V(D)J recombination. AB - We previously demonstrated that DNA molecules with double-strand breaks at variable-(diversity)-joining [V(D)J] recombination signal sequences are relatively abundant in mouse thymocytes. This abundance strongly suggests that the mechanism of V(D)J recombination involves double-strand cleavage at recombination signals. As a first step toward understanding the mechanism of cleavage, we used a sensitive PCR assay to characterize the structure of one class of cleavage products, the signal ends, in detail. Here we demonstrate that most of these ends are blunt and terminate in 5' phosphoryl groups. Virtually all of the flush signal ends are full length. A minor subpopulation of broken ends terminates in short single-strand extensions. We have found no evidence for covalent DNA-protein linkages involving the signal ends. These data allow further refinement of the double-strand cleavage model for V(D)J recombination. PMID- 8248172 TI - Cloning of glycoprotein D cDNA, which encodes the major subunit of the Duffy blood group system and the receptor for the Plasmodium vivax malaria parasite. AB - cDNA clones encoding the major subunit of the Duffy blood group were isolated from a human bone marrow cDNA library using a PCR-amplified DNA fragment encoding an internal peptide sequence of glycoprotein D (gpD) protein. The open reading frame of the 1267-bp cDNA clone indicated that gpD protein was composed of 338 amino acids, predicting a M(r) of 35,733, which was the same as a deglycosylated gpD protein. Portions of the predicted amino acid sequence, matched with six CNBr/pepsin peptides obtained from affinity-purified gpD protein. In ELISA analysis, an anti-Duffy murine monoclonal antibody reacted with a synthetic peptide deduced from the cDNA clone. Hydropathy analysis suggested the presence of 9 membrane-spanning alpha-helices. In bone marrow RNA blot analysis, the gpD cDNA detected a 1.27-kb mRNA in Duffy-positive but not in Duffy-negative individuals. It also identified the same size mRNA in adult kidney, adult spleen, and fetal liver; in brain, it detected a prominent 8.5-kb and a minor 2.2-kb mRNA. In Southern blot analysis, gpD cDNA identified a single gene in Duffy positive and -negative individuals. Duffy-negative individuals, therefore, have the gpD gene, but it is not expressed in bone marrow. The same or a similar gene is active in adult kidney, adult spleen, and fetal liver of Duffy-positive individuals. Whether this is true in Duffy-negative individuals remains to be demonstrated. A GenBank sequence search yielded a significant protein sequence homology to human and rabbit interleukin-8 receptors. PMID- 8248173 TI - Efficacy of beta 1-adrenergic receptors is lower than that of beta 2-adrenergic receptors. AB - We investigated the relative activity at which fully occupied human beta 1- and beta 2-adrenergic receptors (beta 1AR and beta 2AR) activate the stimulatory G protein (Gs)/adenylyl cyclase (AC) system in isolated membranes. The receptors were cloned and coexpressed in permanent cell lines at beta 1/beta 2 ratios that varied from 1:2 to 3:1 and at total receptor abundance that ranged from 8 to 2200 fmol/mg of membrane protein. Cell lines expressing beta 1AR or beta 2AR alone were also obtained. Competitive inhibition of isoproterenol-stimulated AC activity by the beta 2-selective antagonist ICI 118551 showed in all cases that maximal stimulation elicited by beta 1AR was lower than when it was elicited by equivalent densities of beta 2AR. This was especially noticeable at limiting concentrations of receptor, where the beta 1AR-mediated effect was < 10% of that mediated by beta 2AR. At receptor concentrations > 1000 fmol/mg of protein, stimulation by beta 2AR appeared to reach a maximum, while stimulation by beta 1AR continued to increase, so that at 3200 fmol/mg, beta 1AR-stimulated activity was 80% of beta 2AR-stimulated activity. It is clear that the degree to which a given receptor system is able to activate the Gs/AC system depends not only on its abundance but also on an activity parameter determined by the nature of the receptor, which we refer to as receptor efficacy. For human beta ARs, this efficacy parameter is much lower for the beta 1 subtype than for its beta 2 counterpart. The more effective stimulation of AC through beta 2AR than through beta 1AR is an inherent property of the receptor and not the cell in which it is expressed. PMID- 8248174 TI - Acetalins: opioid receptor antagonists determined through the use of synthetic peptide combinatorial libraries. AB - A synthetic peptide combinatorial library made up of 52,128,400 hexapeptides, each having an acetyl group at the N terminus and an amide group on the C terminus, was screened to find compounds able to displace tritiated [D Ala2,MePhe4,Gly-ol5]enkephalin from mu opioid receptor binding sites in crude rat brain homogenates. Individual peptides with mu receptor affinity were found using an iterative process for successively determining the most active peptide mixtures. Upon completion of this iterative process, the three peptides with the highest affinity were Ac-RFMWMT-NH2, Ac-RFMWMR-NH2, and Ac-RFMWMK-NH2. These peptides showed high affinity for mu and kappa 3 opioid receptors, somewhat lower affinity for delta receptors, weak affinity for kappa 1 receptors, and no affinity for kappa 2 receptors. They were found to be potent mu receptor antagonists in the guinea pig ileum assay and relatively weak antagonists in the mouse vas deferens assay. These peptides represent a class of opioid receptor ligands that we have termed acetalins (acetyl plus enkephalin). PMID- 8248175 TI - Role of dimerization in yeast aspartyl-tRNA synthetase and importance of the class II invariant proline. AB - Cytoplasmic aspartyl-tRNA synthetase (AspRS; EC 6.1.1.12) from yeast is, as are most class II synthetases, an alpha 2 dimer. The only invariant amino acid in signature motif 1 of this class is Pro-273; this residue is located at the dimer interface. To understand the role of Pro-273 in the conserved dimeric configuration, we tested the effect of a Pro-273-->Gly (P273G) substitution on the catalytic properties of homo- and heterodimeric AspRS. Heterodimers of AspRS were produced in vivo by overexpression of their respective subunit variants from plasmid-encoded genes and purified to homogeneity in one HPLC step. The homodimer containing the P273G shows an 80% inactivation of the enzyme and an affinity decrease for its cognate tRNA(Asp) of one order of magnitude. The P273G-mutated subunit recovered wild-type enzymatic properties when associated with a native subunit or a monomer otherwise inactivated having an intact dimeric interface domain. These results, which can be explained by the crystal structure of the native enzyme complexed with its substrates, confirm the structural importance of Pro-273 for dimerization and clearly establish the functional interdependence of the AspRS subunits. More generally, the dimeric conformation may be a structural prerequisite for the activity of mononucleotide binding sites constructed from antiparallel beta strands. PMID- 8248176 TI - Yeast precursor mRNA processing protein PRP19 associates with the spliceosome concomitant with or just after dissociation of U4 small nuclear RNA. AB - During assembly of the spliceosome, the U4 small nuclear RNA (snRNA) interacts with the spliceosome as a preformed U4/U6-U5 triple small nuclear ribonucleoprotein (snRNP) complex. Subsequently, U4 becomes loosely associated with the spliceosome, whereas U5 and U6 remain tightly associated, suggesting unwinding of the U4/U6 duplex. We show that this step of the assembly process can be blocked by limiting the ATP concentration in the splicing reaction. We also show that the yeast precursor mRNA processing protein PRP19 becomes associated with the spliceosome during this transition. Thus, PRP19 may function in this step of spliceosome assembly. PMID- 8248177 TI - Phosducin-like protein: an ethanol-responsive potential modulator of guanine nucleotide-binding protein function. AB - Acute and chronic exposure to ethanol produces specific changes in several signal transduction cascades. Such alterations in signaling are thought to be a crucial aspect of the central nervous system's adaptive response, which occurs with chronic exposure to ethanol. We have recently identified and isolated several genes whose expression is specifically induced by ethanol in neural cell cultures. The product of one of these genes has extensive sequence homology to phosducin, a phosphoprotein expressed in retina and pineal gland that modulates trimeric guanine nucleotide-binding protein (G protein) function by binding to G protein beta gamma subunits. We identified from a rat brain cDNA library an isolate encoding the phosducin-like protein (PhLP), which has 41% identity and 65% amino acid homology to phosducin. PhLP cDNA is expressed in all tissues screened by RNA blot-hybridization analysis and shows marked evolutionary conservation on Southern hybridization. We have identified four forms of PhLP cDNA varying only in their 5' ends, probably due to alternative splicing. This 5' end variation generates two predicted forms of PhLP protein that differ by 79 aa at the NH2 terminus. Treatment of NG108-15 cells for 24 hr with concentrations of ethanol seen in actively drinking alcoholics (25-100 mM) causes up to a 3-fold increase in PhLP mRNA levels. Induction of PhLP by ethanol could account for at least some of the widespread alterations in signal transduction and G-protein function that are known to occur with chronic exposure to ethanol. PMID- 8248178 TI - beta-Amyloid-(1-42) is a major component of cerebrovascular amyloid deposits: implications for the pathology of Alzheimer disease. AB - Reinvestigation of the chemical structure of beta-amyloid peptide (A beta) deposits in the vascular tissue of Alzheimer disease brains revealed that the 42 residue form A beta-(1-42), rather than the more soluble A beta-(1-40) form, is the predominant peptide. Following removal of the surrounding tissue with SDS and collagenase, A beta was solubilized in formic acid and purified by Superose 12 chromatography. Peptides generated by enzymatic and chemical digestion of the A beta were purified by HPLC and characterized by amino acid analysis, sequence analysis, and mass spectrometry. In the leptomeningeal vessels, the average ratio of A beta-(1-42)/A beta-(1-40) was 58:42, whereas in the parenchymal vessels this ratio was 75:25. Interestingly, vascular A beta contains considerably less isomerized and racemized aspartyl residues than does neuritic plaque A beta, suggesting that the vascular amyloid is "younger." The discrete nature of the bands and spherical deposits of A beta associated with arterioles and capillaries, respectively, suggests that this amyloid arises from the vascular tissue itself. Increasing A beta deposition appears to lead to the distortion and occlusion of capillaries, which may contribute significantly to the pathology of Alzheimer disease. PMID- 8248179 TI - Brn-3.0: a POU-domain protein expressed in the sensory, immune, and endocrine systems that functions on elements distinct from known octamer motifs. AB - Characterization of Brn-3.0 and identification of a highly related member (Brn 3.1) of the class IV POU-domain family suggest potential roles of Brn-3.0 in the development of retinal ganglion cells and sensory neurons, as well as potential roles in the pituitary gland and the immune system. Brn-3.0 is expressed in the pituitary gland and in a corticotroph cell line. A functional DNA response element has been identified in the proopiomelanocortin promoter. In contrast to previously described mammalian POU-domain proteins, Brn-3.0 binds relatively ineffectively to known octamer DNA motifs, but instead binds with high affinity to a distinct set of DNA elements, functioning as a transcriptional activator. Brn-3.0, Brn-3.1, and the Drosophila tI-POU share an N-terminal region of homology, referred to as the "POU-IV box," which is similar to a conserved functional domain in the c-myc gene family. PMID- 8248180 TI - Heterotypic and homotypic associations between ezrin and moesin, two putative membrane-cytoskeletal linking proteins. AB - Ezrin and moesin are components of actin-rich cell surface structures that are thought to function as membrane-cytoskeletal linking proteins. Here we show that a stable complex of ezrin and moesin can be isolated from cultured cells by immunoprecipitation with specific antibodies. The capacity of these two proteins to interact directly was confirmed with a blot-overlay procedure in which biotin tagged proteins in solution were incubated with immobilized binding partners. In addition to the heterotypic association of ezrin and moesin, homotypic binding of ezrin to ezrin and of moesin to moesin was also demonstrated in vitro. These results suggest mechanisms by which ezrin and moesin might participate in dynamic aspects of cortical cytoskeletal structure. PMID- 8248181 TI - Highly identical cassettes of gene regulatory elements, genomically repetitive and present in RNA. AB - A region in the first intron of a metallothionein-encoding gene of the sea urchin Strongylocentrotus purpuratus (SpMTA gene) regulates its 5' promoter activity. Within this region is a 290-bp cassette of six sequence motifs that are present in other genes in this species and posited to operate as regulatory elements. The cassette, present at high multiplicity in the genome, was used to screen genomic DNA clones. Of these, six diverse individuals were partially sequenced and found to have segments 94% identical to the 290-bp cassette in the SpMTA gene. Their next 80 bp diverged from the SpMTA sequence but were highly identical among the six non-SpMTA clones and contained an additional regulatory motif. These diverse clones thus contained 370-bp cassettes of an overall 94% sequence identity and an apparent content of seven regulatory elements. The regulatory cassettes were transposon-like, insofar as the termini of the highly identical regions consisted of 24- to 25-bp inverted repeats, bracketed by 6- to 9-bp direct repeats in the divergent regions. In addition to being in transcripts of the SpMTA intron, the cassette was found in other sea urchin embryo poly(A)+ RNAs, in eggs and embryos, and enriched in pluteus ectoderm. The cassette sequence was present in moderate abundance in transcripts in both sense and antisense orientation. We report the presence of a transposon-like cassette of regulatory elements that is also represented in RNA, which potentially could function differently from previously described transposons. PMID- 8248182 TI - Cell growth and lambda phage development controlled by the same essential Escherichia coli gene, ftsH/hflB. AB - The lambda phage choice between lysis and lysogeny is influenced by certain host functions in Escherichia coli. We found that the frequency of lambda lysogenization is markedly increased in the ftsH1 temperature-sensitive mutant. The ftsH gene, previously shown to code for an essential inner membrane protein with putative ATPase activity, is identical to hflB, a gene involved in the stability of the phage cII activator protein. The lysogenic decision controlled by FtsH/HflB is independent of that controlled by the protease HflA. Overproduction of FtsH/HflB suppresses the high frequency of lysogenization in an hflA null mutant. The FtsH/HflB protein, which stimulates cII degradation, may be a component of an HflA-independent proteolytic pathway, or it may act as a chaperone, maintaining cII in a conformation subject to proteolysis via such a pathway. Suppressor mutations of ftsH1 temperature-sensitive lethality, located in the fur gene (coding for the ferric uptake regulator), did not restore FtsH/HflB activity with respect to lambda lysogenization. PMID- 8248183 TI - The Escherichia coli hflA locus encodes a putative GTP-binding protein and two membrane proteins, one of which contains a protease-like domain. AB - The hflA (high frequency of lysogenization) locus of Escherichia coli governs the lysis-lysogeny decision of bacteriophage lambda by controlling stability of the phage cII protein. hflA contains three genes, hflX, hflK, and hflC, encoding polypeptides of 50, 46, and 37 kDa, respectively. We have determined the nucleotide sequence of 3843 base pairs containing hflA and have found three large open reading frames corresponding to hflX, hflK, and hflC. HflX contains the three sequence motifs typical of GTP-binding proteins and appears to be a member of a distinct family of putative GTPases. HflC and HflK appear to be integral membrane proteins which show some similarity to each other and to a human membrane protein. The C-terminal region of HflC contains a domain resembling the catalytic domain of ClpP, a bacterial ATP-dependent protease. We hypothesize that HflK and HflC constitute a distinct membrane-bound protease whose activity may be modulated by HflX GTPase. PMID- 8248184 TI - Gastric DNA-binding proteins recognize upstream sequence motifs of parietal cell specific genes. AB - Polymerase chain reaction amplification of cDNA from pig gastric mucosa demonstrated the presence of zinc-finger proteins called GATA-GT1, GATA-GT2, and GATA-GT3, each having zinc-finger sequences similar to previously characterized GATA-binding proteins. Subsequently, full-length cDNAs of GATA-GT1 and GATA-GT2 were obtained from rat stomach. The zinc-finger domains of GATA-GT1 and -GT2 were 66-86% identical on the amino acid level with each other and with other GATA binding proteins. Potential protein kinase phosphorylation sites were present in the zinc-finger region. In contrast, regions outside the zinc fingers shared significantly lower similarities. GATA-GT2 was found to bind to the upstream sequence of the H+/K(+)-ATPase beta gene and to a sequence containing the GATA motif. GATA-GT1 and -GT2 were expressed predominantly in the gastric mucosa and at much lower levels in the intestine (GATA-GT2, also in testis), their tissue distributions being distinct from those of GATA-1, -2, or -3. These results clearly suggest that GATA-GT1 and GATA-GT2 are involved in gene regulation specifically in the gastric epithelium and represent two additional members of the GATA-binding protein family. PMID- 8248185 TI - Rapid assembly of the bacteriophage T4 core replication complex on a linear primer/template construct. AB - DNA synthesis on a primed DNA substrate by bacteriophage T4 requires the assembly of a core replication complex consisting of the T4 DNA polymerase, a single stranded binding protein (32 protein), and the accessory proteins 44/62 and 45. In this paper, we demonstrate the successful assembly of this core complex on a short linear primer/template system at levels of accessory proteins equivalent to the concentration of primer 3' ends. The key to this assembly is the presence of streptavidin molecules bound at each end of the DNA substrate via biotin moieties incorporated into the template strand. Streptavidin serves to block the ends of the primer/template, thus preventing translocation of the accessory proteins away from the site of assembly and their subsequent dissociation from the ends of the primer/template. Complex assembly on this substrate requires ATP and the presence of both the 44/62 and 45 proteins. The time required for assembly of a full enzyme equivalent of complex in our system is approximately 2 s. PMID- 8248186 TI - Evidence for linkage of the apolipoprotein A-II locus to plasma apolipoprotein A II and free fatty acid levels in mice and humans. AB - Although it has been hypothesized that the synteny between mouse and human genes provides an approach to the localization of genes that determine quantitative traits in humans, this has yet to be demonstrated. We tested this approach with two quantitative traits, plasma apolipoprotein A-II (apoAII) and free fatty acid (FFA) levels. ApoAII is the second most abundant protein of high density lipoprotein particles, but its function remains largely unknown. We now show that, in a backcross between strains Mus spretus and C57BL/6J, apoAII levels correlate with plasma FFA concentrations on both chow (P < 0.0001) and high-fat (P < 0.0003) diets and that apoAII levels are linked to the apoAII gene (P < 0.0002). To test whether variations of the apoAII gene influence plasma lipid metabolism in humans, we studied 306 individuals in 25 families enriched for coronary artery disease. The segregation of the apoAII gene was followed by using an informative simple sequence repeat in the second intron of the gene and two nearby genetic markers. Robust sib-pair linkage analysis was performed on members of these families using the SAGE linkage programs. The results suggest linkage between the human apoAII gene and a gene controlling plasma apoAII levels (P = 0.03). Plasma apoAII levels were also significantly correlated with plasma FFA levels (P = 0.007). Moreover, the apoAII gene exhibited linkage with a gene controlling FFA levels (P = 0.003). Evidence for nonrandom segregation was seen with markers as far as 6-12 centimorgans from the apoAII structural locus. These data provide evidence, in two species, that the apoAII gene is linked to a gene that controls plasma apoAII levels and that apoAII influences, by an unknown mechanism, plasma FFA levels. The results illustrate the utility of animal studies for analysis of complex traits. PMID- 8248187 TI - A receptor tyrosine kinase found in breast carcinoma cells has an extracellular discoidin I-like domain. PMID- 8248188 TI - Proteins, radicals, isotopes, and mutants in photosynthetic oxygen evolution. PMID- 8248189 TI - RNA enzyme-directed gene therapy. PMID- 8248190 TI - Positron emission tomography as a tool to study human vision and attention. PMID- 8248191 TI - Conventional protein coding genes in the Drosophila Y chromosome: is the puzzle of the fertility gene function solved? PMID- 8248192 TI - What determines where alpha-helices begin and end? PMID- 8248193 TI - Caveolae: where incoming and outgoing messengers meet. AB - Plasmalemmal caveolae were first identified as an endocytic compartment in endothelial cells, where they appear to move molecules across the cell by transcytosis. More recently, they have been found to be sites where small molecules are concentrated and internalized by a process called potocytosis. A growing body of biochemical and morphological evidence indicates that a variety of molecules known to function directly or indirectly in signal transduction are enriched in caveolae. This raises the possibility that a third function for caveolae is to process hormonal and mechanical signals for the cell. Insights gained from studying potocytosis suggest several different ways that this membrane specialization might function to integrate incoming and outgoing cellular messages. PMID- 8248194 TI - Different perceptual tasks performed with the same visual stimulus attribute activate different regions of the human brain: a positron emission tomography study. AB - To investigate the processing of visual form in human cerebral cortex, we used the PET (positron emission tomography) activation technique to compare the human brain regions that are involved in a visual detection task and two orientation discrimination tasks: the temporal same-different (TSD) task, which includes a short-term memory component, and the identification (ID) task, which is without this component. As a control task we used passive viewing. Stimuli were identical in all four tasks. Subtraction of passive viewing from detection showed that the detection task activates early visual cortical regions (areas 17/18) as well as several motor brain regions, while decreasing activity in several higher order frontal, temporal, and parietal regions. Comparing the ID task to the detection task revealed no further visual cortical activation, while comparison of the TSD task to the detection task revealed an activation of several right visual cortical regions, one of which remained significant after the subtraction of ID from TSD (right area 19). These experiments demonstrate the task dependence of visual processing, even for very closely related tasks, and the localization of the temporal comparison component involved in orientation discrimination in human area 19. PMID- 8248195 TI - A functional model for the role of cytochrome b559 in the protection against donor and acceptor side photoinhibition. AB - A quinone-independent photoreduction of the low potential form of cytochrome b559 has been studied using isolated reaction centers of photosystem II. Under anaerobic conditions, the cytochrome can be fully reduced by exposure to strong illumination without the addition of any redox mediators. Under high light conditions, the extent and rate of the reduction is unaffected by addition of the exogenous electron donor Mn2+ and, during this process, no irreversible damage occurs to the reaction center. However, prolonged illumination in strong light brings about irreversible bleaching of chlorophyll, indicative of photoinhibitory damage. When the cytochrome is fully reduced and excess Mn2+ is present, the effect of moderate light is to facilitate the photoaccumulation of reduced pheophytin. The dark reoxidation of the reduced cytochrome is very slow under anaerobic conditions but significantly speeded up on addition of oxidized 2,5 dibromo-3-methyl-6-isopropyl-p-benzoquinone. From these results it is suggested that the low potential form of cytochrome b559 can accept electrons directly from reduced pheophytin and in so doing help to protect the reaction center against acceptor side photoinhibition as suggested by Nedbal et al. [Nedbal, J., Samson, G. & Whitmarsh, J. (1992) Proc. Natl. Acad. Sci. USA 89, 7929-7933]. This conclusion has been incorporated into a model that further suggests that in its high potential form the cytochrome primarily acts to protect against donor side photoinhibition due to increased lifetime of highly oxidized species as previously proposed by Thompson and Brudvig [Thompson, L. & Brudvig, G. W. (1988) Biochemistry 27, 6653-6658]. The particular feature of our scheme is that it incorporates reversible interconversion between the two redox forms so as to protect against either type of photoinhibition. PMID- 8248196 TI - Raf-1 forms a stable complex with Mek1 and activates Mek1 by serine phosphorylation. AB - Recombinant Mek1 and Raf-1 proteins produced in Sf9 cells undergo a tight association both in vivo and in vitro, which apparently does not depend on additional factors or the kinase activity of Mek1 or Raf-1. The complex can be disrupted by two polyclonal antibodies raised against Raf-1 peptides. Coinfection with Raf-1 activates Mek1 > 150-fold, and coinfection with Raf-1 and Mek1 activates Erk1 approximately 90-fold. The activation of Mek1 by Raf-1 involves only serine phosphorylation, which is directly proportional to the extent of Mek1 activation. Phosphopeptide maps suggest a single Raf-1 phosphorylation site on mek1. PMID- 8248197 TI - Phosphorylation of the c-Fos transrepression domain by mitogen-activated protein kinase and 90-kDa ribosomal S6 kinase. AB - Phosphorylation of the C terminus of c-Fos has been implicated in serum response element-mediated repression of c-fos transcription after its induction by serum growth factors. The growth-regulated enzymes responsible for this phosphorylation in early G1 phase of the cell cycle and the sites of phosphorylation have not been identified. We now provide evidence that two growth-regulated, nucleus- and cytoplasm-localized protein kinases, 90-kDa ribosomal S6 kinase (RSK) and mitogen activated protein kinase (MAP kinase), contribute to the serum-induced phosphorylation of c-Fos. The major phosphopeptides derived from biosynthetically labeled c-Fos correspond to phosphopeptides generated after phosphorylation of c Fos in vitro with both RSK and MAP kinase. The phosphorylation sites identified for RSK (Ser-362) and MAP kinase (Ser-374) are in the transrepression domain. Cooperative phosphorylation at these sites by both enzymes was observed in vitro and reflected in vivo by the predominance of the peptide phosphorylated on both sites, as opposed to singly phosphorylated peptides. This study suggests a role for nuclear RSK and MAP kinase in modulating newly synthesized c-Fos phosphorylation and downstream signaling. PMID- 8248198 TI - Endogenous nitrogen oxides and bronchodilator S-nitrosothiols in human airways. AB - Recent discoveries suggesting essential bioactivities of nitric oxide (NO.) in the lung are difficult to reconcile with the established pulmonary cytotoxicity of this common air pollutant. These conflicting observations suggest that metabolic intermediaries may exist in the lung to modulate the bioactivity and toxicity of NO.. We report that S-nitrosothiols (RS-NO), predominantly the adduct with glutathione, are present at nano- to micromolar concentrations in the airways of normal subjects and that their levels vary in different human pathophysiologic states. These endogenous RS-NO are long-lived, potent relaxants of human airways under physiological O2 concentrations. Moreover, RS-NO form in high concentrations upon administration of NO. gas. Nitrite (10-20 microM) is found in airway lining fluid in concentrations linearly proportional to leukocyte counts, suggestive of local NO. metabolism. NO. itself was not detected either free in solution or in complexes with transition metals. These observations may provide insight into the means by which NO. is packaged in biological systems to preserve its bioactivity and limit its potential O2-dependent toxicity and suggest an important role for NO. in regulation of airway luminal homeostasis. PMID- 8248199 TI - Theoretical basis for separation of multiple linked gene effects in mapping quantitative trait loci. AB - It is now possible to use complete genetic linkage maps to locate major quantitative trait loci (QTLs) on chromosome regions. The current methods of QTL mapping (e.g., interval mapping, which uses a pair or two pairs of flanking markers at a time for mapping) can be subject to the effects of other linked QTLs on a chromosome because the genetic background is not controlled. As a result, mapping of QTLs can be biased, and the resolution of mapping is not very high. Ideally when we test a marker interval for a QTL, we would like our test statistic to be independent of the effects of possible QTLs at other regions of the chromosome so that the effects of QTLs can be separated. This test statistic can be constructed by using a pair of markers to locate the testing position and at the same time using other markers to control the genetic background through a multiple regression analysis. Theory is developed in this paper to explore the idea of a conditional test via multiple regression analysis. Various properties of multiple regression analysis in relation to QTL mapping are examined. Theoretical analysis indicates that it is advantageous to construct such a testing procedure for mapping QTLs and that such a test can potentially increase the precision of QTL mapping substantially. PMID- 8248200 TI - Transcriptional organization of a 450-kb region of the human X chromosome in Xq28. AB - In this paper, we report the transcriptional organization of a 450-kb gene cluster in Xq28, flanked by the glucose-6-phosphate dehydrogenase and the color vision genes. CpG islands previously identified and mapped to distal Xq28 have helped in construction of a continuous contig of cosmids and in identification of cDNAs corresponding to eight transcripts. Thirteen to 16 small genes with CpG islands are clustered in a region of 250-300 kb. Many are highly expressed in muscle or brain and may be the genes responsible for muscle or neurological disorders mapped to distal Xq28. Our analysis indicates that, in this region of the genome, genes not related in sequence are organized in transcriptional domains of 100 kb and that this organization may be important for establishing and regulating gene expression in relation to tissue distribution and X chromosome inactivation. PMID- 8248201 TI - Induction of apoptosis in cerebellar granule neurons by low potassium: inhibition of death by insulin-like growth factor I and cAMP. AB - High levels of extracellular K+ ensure proper development and prolong survival of cerebellar granule neurons in culture. We find that when switched from a culture medium containing high K+ (25 mM) to one containing a low but more physiological K+ concentration (5 mM), differentiated granule neurons degenerate and die. Death induced by low K+ is due to apoptosis (programmed cell death), a form of cell death observed extensively in the developing nervous system and believed to be necessary for proper neurogenesis. The death process is accompanied by cleavage of genomic DNA into internucleosome-sized fragments, a hallmark of apoptosis. Inhibitors of transcription and translation suppress apoptosis induced by low K+, suggesting the necessity for newly synthesized gene products for activation of the process. Death can be prevented by insulin-like growth factor I but not by several other growth/neurotrophic factors. cAMP but not the protein kinase C activator phorbol 12-myristate 13-acetate can also support survival in low K+. In view of the large numbers of granule neurons that can be homogeneously cultured, our results offer the prospect of an excellent model system to study the mechanisms underlying apoptosis in the central nervous system and the suppression of this process by survival factors such as insulin-like growth factor I. PMID- 8248202 TI - On the puzzling arrangement of the asymmetric MalT-binding sites in the MalT dependent promoters. AB - The MalT-dependent promoters of the enterobacteria belong to a small family of positively regulated prokaryotic promoters in which the activator protein recognizes short asymmetric nucleotide sequences present in several locations and orientations. We demonstrate that active MalT-dependent semisynthetic promoters can be constructed by using a synthetic decanucleotide as the MalT site and random nucleotide sequences as connecting sequences, provided that the location and orientation of the sites are the same as in natural MalT-dependent promoters. Strikingly, the induced level of promoter activity and the induction factor of each semisynthetic promoter are identical to those of its natural counterpart, in spite of considerable differences in their nucleotide sequences. The study of these semisynthetic promoters confirms the importance of the structural motif formed by two MalT sites in a direct repeat. This motif is involved in promoter activation either alone or in conjunction with a third MalT site, proximal with respect to the transcription start site. In this latter configuration, the promoters are active irrespective of the orientation of the repeat, and they retain at least some activity when the distance between the repeat and the proximal site is increased, provided that the alignment along the axis of the helix is conserved. PMID- 8248203 TI - Purification and characterization of angiotensin II AT2 receptors from neonatal rat kidney. AB - Angiotensin II (Ang II) AT2 receptors were purified 40,000-fold to a nearly homogeneous state after solubilization from neonatal rat kidney membranes with 3 [(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propane-sulfonic acid. Comparable IC50 values for the soluble extract (0.32 nM) and membranes (0.31 nM) were obtained by competition curves with 125I-labeled CGP42112, a selective AT2 ligand. Binding to AT2 receptors in the soluble extract was not sensitive to dithiothreitol. AT2 receptors were further purified by gel filtration and a CGP42112 Sepharose affinity column. Ang II AT2 receptors were selectively eluted with 5 microM CGP42112 at 4 degrees C, and a single band with an apparent molecular mass of 71 kDa was obtained after SDS/PAGE. Two-dimensional electrophoresis confirmed the purity of the protein and an isoelectric point of 5.3-5.5 was obtained. A highly selective elution of the AT2 receptors from the affinity column was performed with 5 nM 125I-labeled CGP42112 at room temperature after the column was treated with 1 microM losartan in the presence of high salt. After cross-linking, a major labeled protein with similar molecular mass and isoelectric point was obtained. Dissociation of the radiolabeled protein was insensitive to losartan but was enhanced by CGP42112, PD123177, Ang II, and [Sar1]Ang II. In summary, Ang II AT2 receptors were purified by CGP42112 affinity chromatography and selective elution and retain the pharmacological specificity of particulate receptors. PMID- 8248204 TI - cDNAs encoding the large subunit of human replication factor C. AB - Replication factor C (RFC) is a multisubunit, DNA polymerase accessory protein required for the coordinated synthesis of both DNA strands during simian virus 40 DNA replication in vitro. Previous studies have shown that RFC is a DNA-dependent ATPase that binds in a structure-specific manner to the 3' end of a primer hybridized to a template DNA, an activity thought intrinsic to the 140-kDa component of this multisubunit complex. Here, the isolation and analysis of cDNAs encoding this subunit is described. Analysis of the full-length coding sequence revealed an open reading frame of 3.4 kb, encoding an 1148-amino acid protein with a predicted molecular mass of 130 kDa. A putative ATP-binding motif was observed that is similar to a motif in several of the smaller subunits of RFC and in functionally homologous replication factors of bacterial and viral origin. A "DEAD" box is also conserved among these proteins. The predicted protein shows significant identity with a DNA-binding protein of murine origin (B. Luckow, P. Lichter, and G. Schutz, personal communication). Regions of similarity were also seen between the amino acid sequences of the 140-kDa subunit of RFC, poly(ADP ribose) polymerase, and bacterial DNA ligases--possibly representing a conserved structural feature of these proteins that bind similar DNA substrates. PMID- 8248205 TI - Autoregulation of the Escherichia coli heat shock response by the DnaK and DnaJ heat shock proteins. AB - All organisms respond to various forms of stress, including heat shock. The heat shock response has been universally conserved from bacteria to humans. In Escherichia coli the heat shock response is under the positive transcriptional control of the sigma 32 polypeptide and involves transient acceleration in the rate of synthesis of a few dozen genes. Three of the heat shock genes--dnaK, dnaJ, and grpE--are special because mutations in any one of these lead to constitutive levels of heat shock gene expression, implying that their products negatively autoregulate their own synthesis. The DnaK, DnaJ, and GrpE proteins have been known to function in various biological situations, including bacteriophage lambda replication. Here, we report the formation of an ATP hydrolysis-dependent complex of DnaJ, sigma 32, and DnaK proteins in vitro. This DnaJ-sigma 32-DnaK complex has been seen under different conditions, including glycerol gradient sedimentation and co-immunoprecipitation. The DnaK and DnaJ proteins in the presence of ATP can interfere with the efficient binding of sigma 32 to the RNA polymerase core, and are capable of disrupting a preexisting sigma 32-RNA polymerase complex. Our results suggest a possible mechanism for the autoregulation of the heat shock response. PMID- 8248206 TI - A difference Fourier-transform infrared study of two redox-active tyrosine residues in photosystem II. AB - Photosystem II, the photosynthetic water-oxidizing complex, contains two redox active tyrosine residues. Although current models suggest that these tyrosines are located in symmetric positions in the reaction center, there are functional differences between them. To elucidate those structural factors that give rise to this functional asymmetry, we have used difference Fourier-transform infrared spectroscopy to obtain the vibrational difference spectrum associated with the oxidation of each of these redox-active residues. Isotopic labeling was employed to definitively assign vibrational lines to the redox-active tyrosines. This work has shown that the vibrational spectra of the two redox-active species are significantly different from each other. This result suggests that the structure of the redox-active residue helps to determine its role in electron transfer in the reaction center. PMID- 8248207 TI - Production of immortalized distal respiratory epithelial cell lines from surfactant protein C/simian virus 40 large tumor antigen transgenic mice. AB - Murine lung epithelial (MLE) cell lines representing the distal bronchiolar and alveolar epithelium were produced from lung tumors generated in transgenic mice harboring the viral oncogene simian virus 40 (SV40) large tumor antigen under transcriptional control of a promoter region from the human surfactant protein C (SP-C) gene. The cell lines exhibited rapid growth, lack of contact inhibition, and an epithelial cell morphology for 30-40 passages in culture. Microvilli, cytoplasmic multivesicular bodies, and multilamellar inclusion bodies (morphologic characteristics of alveolar type II cells) were detected in some of the MLE cell lines by electron microscopic analysis. The MLE cells also maintained functional characteristics of distal respiratory epithelial cells including the expression of surfactant proteins and mRNAs and the ability to secrete phospholipids. Expression of the exogenous SV40 large tumor antigen gene was detected in all of the generated cell lines. The SP-C/SV40 large tumor antigen transgenic mice and the MLE cell lines will be useful for the study of pulmonary surfactant production and regulation as well as lung development and tumorigenesis. PMID- 8248208 TI - Altered regulation of G1 cyclins in senescent human diploid fibroblasts: accumulation of inactive cyclin E-Cdk2 and cyclin D1-Cdk2 complexes. AB - Senescent human diploid fibroblasts are unable to enter S phase in response to mitogenic stimulation. One of the key deficiencies in mitogen-stimulated senescent cells is their failure to phosphorylate the retinoblastoma protein, which acts as an inhibitor of entry into S phase in its unphosphorylated form. Recent data suggest that cyclin-dependent kinases (Cdks) regulated by G1 cyclins (D type and E) are responsible for the primary phosphorylation of the retinoblastoma protein prior to the G1/S boundary. Surprisingly, we found 10- to 15-fold higher constitutive amounts of both cyclin E and cyclin D1 in senescent cells compared to quiescent early-passage cells. Nevertheless, cyclin E associated kinase activity in senescent cells was very low and did not increase significantly upon mitogenic stimulation even though cyclin E-Cdk2 complexes were abundant. In contrast to early-passage cells in late G1 phase, senescent cells contained mainly underphosphorylated cyclin E and proportionally more unphosphorylated and inactive Cdk2, perhaps accounting for the low kinase activity. We also show that a majority of the Cdk2 in senescent cells, but not in early-passage cells, was complexed with cyclin D1. Cyclin D1-Cdk2 complexes, severalfold enriched in senescent cells, contained exclusively unphosphorylated Cdk2. Amounts of cyclin A, which ordinarily accumulates in S and G2 phases, were extremely low in stimulated senescent cells. We suggest that the failure to activate cyclin E-Cdk2 kinase activity in senescent cells may account for the inability of these cells to phosphorylate the retinoblastoma protein in late G1 phase, which in turn may block the expression of late G1 genes such as cyclin A that are required for entry into S phase. PMID- 8248209 TI - Molecular cloning and centrosomal localization of human caltractin. AB - Caltractin, a 20-kDa calcium-binding protein, was previously purified and cloned at the DNA level from the unicellular green alga Chlamydomonas. It is a structural component of the basal body complex, the major microtubule-organizing center in Chlamydomonas and the functional homolog of the centrosome in the animal cell. Here we report the characterization of a cDNA encoding a human caltractin that shares a high degree of amino acid identity (70%) with its algal counterpart. Caltractin was identified in both HeLa and BHK cells as a 21-kDa polypeptide specifically localized to the centrosome of interphase and mitotic cells. The high level of conservation in the amino acid sequence of caltractin from algae to humans and its association with the major microtubule-organizing center in the cell suggest that caltractin plays a fundamental role in microtubule-organizing center structure and function. PMID- 8248210 TI - Cooperative inhibition of NF-kappa B and Tat-induced superactivation of human immunodeficiency virus type 1 long terminal repeat. AB - Human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR)-regulated gene expression is stimulated independently by the cellular trans-activator NF kappa B and the viral protein Tat. Noncytotoxic concentrations of the drug pentoxifylline (PTX) inhibited interaction of NF-kappa B with its motif and the stimulation of HIV-1 LTR-driven gene expression in Jurkat cells. Tat protein (from a cotransfected Tat-expression vector) also induced activation of HIV-1 LTR driven gene expression. This activation was unaffected by PTX when NF-kappa B sites in the HIV-1 LTR were mutated, suggesting that this drug does not directly influence Tat function, which, however, was inhibited by the Tat-inhibitor Ro 24 7429. Transient reporter gene expression regulated by HIV-1 LTR with wild-type NF kappa B motifs in the presence of Tat protein was 10- to 60-fold higher than in the presence of either of the trans-activators alone, demonstrating superactivation of HIV-1 LTR by the concerted action of both the trans activators. Treatment of cells with either PTX or Ro 24-7429 inhibited this superactivation of the HIV-1 LTR. The inhibitory effect of these two drugs in combination, at concentrations that alone did not significantly influence viral promoter activity, was far more than additive. A cooperative action of PTX (NF kappa B inhibitor) and Ro 24-7429 (Tat inhibitor) on HIV-1 LTR-regulated gene expression is suggested. Concentrations of the drugs that induced maximum inhibition of HIV-1 LTR through their cooperative action are far below cytotoxic levels. Thus, the combination of these two inhibitors could be very effective for anti-HIV therapy. PMID- 8248211 TI - 5-Ethynyluracil (776C85): a potent modulator of the pharmacokinetics and antitumor efficacy of 5-fluorouracil. AB - 5-Ethynyluracil (5-EU, 776C85) is a mechanism-based irreversible inhibitor of dihydropyrimidine dehydrogenase (EC 1.3.1.2), the rate-determining enzyme in 5 fluorouracil (5-FU) catabolism. In the present study, 5-EU was found to be a potent modulator of 5-FU catabolism in mice and rats. Liver extracts prepared up to 6 hr after a 5-EU dose (2 mg/kg) were > 96% inhibited in their ability to catalyze 5-FU degradation. 5-EU treatment increased the elimination t1/2 and the area under the plasma concentration-time curve of 5-FU. 5-FU oral bioavailability was approximately 100% in rats pretreated with 5-EU. Consequently, 5-EU induced a linear relationship between the area under the plasma concentration-time curve and the oral dose of 5-FU. As expected from the preservation of plasma 5-FU, 5-EU potentiated the antitumor activity and the toxicity of 5-FU in two mouse tumor models (Colon 38 and MOPC-315). However, 5-EU potentiated the antitumor activity to a greater degree and thereby increased the therapeutic index of 5-FU 2- to 4 fold. PMID- 8248212 TI - TAP1 alleles in insulin-dependent diabetes mellitus: a newly defined centromeric boundary of disease susceptibility. AB - It has been previously demonstrated that individuals with certain DR alleles have an increased relative risk of developing insulin-dependent diabetes mellitus (IDDM). The disease association is even stronger with certain DQ alleles but there is little association with DP providing a boundary of disease association to the 430 kb between DQ and DP. The recently described TAP (transporter associated with antigen processing) genes have been mapped approximately midway between DP and DQ. Therefore, it was of interest to determine if any TAP alleles were associated with IDDM. In addition to the alleles of TAP1 that have been described, others were identified during this study. Diabetics and normal controls were screened for TAP1 using single-stranded conformational polymorphism and relative risk was determined. In the same population group we have studied extensively in the past, we found a higher association of a TAP1 allele with IDDM than with any single HLA-DP allele but the risk was lower than with HLA DQB1*0302. These data provide new limits for IDDM susceptibility to the 190-kb interval between TAP1 and HLA-DQB1. PMID- 8248213 TI - "BFA bodies": a subcompartment of the endoplasmic reticulum. AB - A specialized region of the endoplasmic reticulum--the BFA body--is defined by the site of accumulation of coatomer when nonclathrin coat protein (COP)-coated vesicle assembly is prevented by the drug brefeldin A (BFA). BFA bodies are formed by part smooth, part rough domains of endoplasmic reticulum that are cis to the classical transitional endoplasmic reticulum and to BFA-induced Golgi remnants. PMID- 8248214 TI - Opiate-like substances in an invertebrate, an opiate receptor on invertebrate and human immunocytes, and a role in immunosuppression. AB - The presence of morphine-like and codeine-like substances was demonstrated in the pedal ganglia, hemolymph, and mantle tissues of the mollusc Mytilus edulis. The pharmacological activities of the endogenous morphine-like material resemble those of authentic morphine. Both substances were found to counteract, in a dose dependent manner, the stimulatory effect of tumor necrosis factor alpha or interleukin 1 alpha on human monocytes and Mytilus immunocytes, when added simultaneously to the incubation medium. The immunosuppressive effect of this opiate material expresses itself in a lowering of chemotactic activity, cellular velocity, and adherence. Codeine mimics the activity of authentic morphine, but only at much higher concentrations. Specific high-affinity receptor sites (mu 3) for morphine have been identified on human monocytes and Mytilus immunocytes. In Mytilus recovering from experimentally induced stress, the return of "altered" immunocytes to a more inactive state appears to be due to a significant rise in the content of morphine-like material in the pedal ganglia and hemolymph at this time. Thus, morphine may have a role in calming or terminating the state of immune alertness. PMID- 8248215 TI - Dominant T-cell-receptor beta chain variable region V beta 14+ clones in juvenile rheumatoid arthritis. AB - The characteristic histopathology and major histocompatibility complex associations in juvenile rheumatoid arthritis suggest an oligoclonal antigen specific T-cell population may be critical to pathogenesis. To test this, we analyzed the T-cell repertoire of a polyarticular HLA-DR4+ juvenile rheumatoid arthritis patient with an aggressive form of disease that required arthrocentesis of the knee joints and early replacement of both hip joints. A comparison of T cell-receptor beta chain variable region (V beta) gene expression in peripheral blood and synovial fluid performed by semiquantitation of cDNA samples amplified by the PCR revealed overexpression of the T-cell-receptor V beta 14 gene family. To determine the nature of V beta 14 overexpression, we sequenced randomly cloned amplification products derived from two synovial fluid, two synovial tissue, and three peripheral blood samples by using a V beta 14/beta chain constant region primer pair. Sequence data showed that the T-cell response in the synovia was oligoclonal. Of four clones found, one was present in all joints examined and persisted over time. This clone accounted for 67% and 74% of all V beta 14+ clones sequenced in two synovial fluid samples and 75% and 40% in two synovial tissue samples. This clone was also found at a lesser frequency in peripheral blood samples. Further studies provided evidence for the presence of oligoclonally expanded populations of T cells utilizing the V beta 14 T-cell receptor in 6 of 27 patients examined. In contrast to the remaining patients studied, 3 with a late onset polyarticular course who exhibited especially marked clonality were characterized by features typical of adult rheumatoid arthritis (IgM rheumatoid factor-positive and HLA-DR4+). These data suggest a role for V beta 14+ T cells in a group of juvenile rheumatoid arthritis patients. PMID- 8248216 TI - Dimer formation by an N-terminal coiled coil in the APC protein. AB - Mutations in the human APC gene are associated with an inherited predisposition to colon cancer. APC codes for polypeptides of approximately 2800 amino acids, with sequence homologies to coiled-coil proteins in the first 900 residues. To determine the oligomerization properties of the APC protein, we used genetic and biochemical approaches to examine the ability of APC fragments to self-associate. A subdomain comprising the first 55 amino acids of APC was found to form a stable, parallel, helical dimer, as expected for a coiled coil. The location of a key dimerization element at the N terminus of the protein supports models in which mutations in APC exert effects through dimerization of the mutant gene products. PMID- 8248217 TI - Genomic binding-site cloning reveals an estrogen-responsive gene that encodes a RING finger protein. AB - Estrogen receptor (ER)-binding fragments were isolated from human genomic DNA by using a recombinant ER protein. Using one of these fragments as a probe, we have identified an estrogen-responsive gene that encodes a putative zinc finger protein. It has a RING finger motif present in a family of apparent DNA-binding proteins and is designated estrogen-responsive finger protein (efp). efp cDNA contains a consensus estrogen-responsive element at the 3' untranslated region that can act as a downstream estrogen-dependent enhancer. Moreover, efp is regulated by estrogen as demonstrated at both the mRNA and the protein level in ER-positive cells derived from mammary gland. These data suggest that efp may represent an estrogen-responsive transcription factor that mediates phenotypic expression of the diverse estrogen action. Thus, the genomic binding-site cloning may be applicable for isolation of the target genes of other transcription factors. PMID- 8248218 TI - Estrogen modulates expression of the glycosyltransferases that synthesize sulfated oligosaccharides on lutropin. AB - The glycoprotein hormone lutropin (LH) bears oligosaccharides terminating with the sequence SO4-4-GalNAc beta 1,4GlcNAc beta 1,2 Man alpha. We have determined that estrogen actively modulates expression of the GalNAc- and sulfotransferases responsible for synthesis of sulfated oligosaccharides on LH alpha and beta subunits. Consequently, terminal glycosylation of LH oligosaccharides with GalNAc 4-SO4 is maintained when LH synthesis and secretion are markedly increased, as occurs during the midcycle surge and following ovariectomy. Maintenance of sulfated oligosaccharides on LH has important biologic consequences because LH circulatory half-life as well as biologic activity at the hormone receptor level are dramatically affected by glycosylation. To our knowledge, regulation of glycosyltransferase levels in response to specific stimuli has not been observed previously, further emphasizing the biologic significance of glycosylation for expression of LH bioactivity in vivo. PMID- 8248219 TI - A fertility region on the Y chromosome of Drosophila melanogaster encodes a dynein microtubule motor. AB - A clone encoding a portion of the highly conserved ATP-binding domain of a dynein heavy-chain polypeptide was mapped to a region of the Drosophila melanogaster Y chromosome. Dyneins are large multisubunit enzymes that utilize the hydrolysis of ATP to move along microtubules. They were first identified as the motors that provide the force for flagellar and ciliary bending. Seven different dynein heavy chain genes have been identified in D. melanogaster by PCR. In the present study, we demonstrate that one of the dynein genes, Dhc-Yh3, is located in Y chromosome region h3, which is contained within kl-5, a locus required for male fertility. The PCR clone derived from Dhc-Yh3 is 85% identical to the corresponding region of the beta heavy chain of sea urchin flagellar dynein but only 53% identical to a cytoplasmic dynein heavy chain from Drosophila. In situ hybridization to Drosophila testes shows Dhc-Yh3 is expressed in wild-type males but not in males missing the kl-5 region. These results are consistent with the hypothesis that the Y chromosome is needed for male fertility because it contains conventional genes that function during spermiogenesis. PMID- 8248220 TI - Visual performance in behaving cats after prenatal unilateral enucleation. AB - Prenatal unilateral enucleation in mammals causes an extensive anatomical reorganization of visual pathways. The remaining eye innervates the entire extent of visual subcortical and cortical areas. Electrophysiological recordings have shown that the retino-geniculate connections are retinotopically organized and geniculate neurones have normal receptive field properties. In area 17 all neurons respond to stimulation of the remaining eye and retinotopy, orientation columns, and direction selectivity are maintained. The only detectable change is a reduction in receptive field size. Are these changes reflected in the visual behavior? We studied visual performance in cats unilaterally enucleated 3 weeks before birth (gestational age at enucleation, 39-42 days). We tested behaviorally the development of visual acuity and, in the adult, the extension of the visual field and the contrast sensitivity. We found no difference between prenatal monocularly enucleated cats and controls in their ability to orient to targets in different positions of the visual field or in their visual acuity (at any age). The major difference between enucleated and control animals was in contrast sensitivity:prenatal enucleated cats present a loss in sensitivity for gratings of low spatial frequency (below 0.5 cycle per degree) as well as a slight increase in sensitivity at middle frequencies. We conclude that prenatal unilateral enucleation causes a selective change in the spatial performance of the remaining eye. We suggest that this change is the result of a reduction in the number of neurones with large receptive fields, possibly due to a severe impairment of the Y system. PMID- 8248221 TI - Isolation of cDNA of an auxin-regulated gene encoding a G protein beta subunit like protein from tobacco BY-2 cells. AB - The addition of 2,4-dichlorophenoxyacetic acid to tobacco BY-2 cells that had been cultured in modified Linsmaier and Skoog medium deprived of auxin for 3 days induced cell division, whereas without 2,4-dichlorophenoxy-acetic acid application, no such induction of cell division was seen. When differential cDNA screening for auxin was done at 4 hr after the addition of 2,4 dichlorophenoxyacetic acid, the cDNA of an auxin-responsive gene designated arcA was isolated. The predicted gene product of arcA is a polypeptide with a M(r) of 35,825. arcA, thus, is a plant hormone-regulated gene that encodes a protein structurally related to the beta subunit of a guanine nucleotide-binding regulatory protein, which is composed of seven repetitive segments of Trp-Asp 40 aa repeats. The possibility that arcA gene products induce cell division is discussed. PMID- 8248222 TI - Drosophila retinal degeneration A gene encodes an eye-specific diacylglycerol kinase with cysteine-rich zinc-finger motifs and ankyrin repeats. AB - The Drosophila visual mutant, carrying the retinal degeneration A gene (rdgA), has photoreceptor cells that degenerate within a week after eclosion. Morphological studies suggested that this mutant harbors abnormalities in membrane turnover of the photoreceptor cells. Biochemically, the rdgA mutant lacks an eye-specific and membrane-associated diacylglycerol kinase (DGK; EC 2.7.1.107) activity in a gene-dosage-dependent manner, suggesting that rdgA gene encodes a DGK. We report the molecular cloning and characterization of a DGK gene, which maps to the rdgA locus. This gene, designated as DGK2, has a single open reading frame that encodes 1454 amino acids. Like porcine DGK, DGK2 has two cysteine-rich zinc-finger motifs as well as a DGK catalytic domain. The DGK2 protein contains four ankyrin-like repeats at the C-terminal region, suggesting that DGK2 is likely anchored to the membrane or cytoskeleton. Northern blot analysis and tissue in situ hybridization to adult sections revealed that DGK2 is expressed exclusively in the adult retina and that the amount of its mRNA is reduced in some of the rdgA mutant alleles. Furthermore, in two rdgA alleles, rdgA1 and rdgA2, nonsense and missense mutations occur within their DGK2 gene, respectively. Thus, we conclude that rdgA encodes an eye-specific DGK, the absence of which leads to rhabdomere degeneration due to defective phospholipid turnover. PMID- 8248223 TI - Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor gene. AB - Estrogen receptor and its ligand, estradiol, have long been thought to be essential for survival, fertility, and female sexual differentiation and development. Consistent with this proposed crucial role, no human estrogen receptor gene mutations are known, unlike the androgen receptor, where many loss of function mutations have been found. We have generated mutant mice lacking responsiveness to estradiol by disrupting the estrogen receptor gene by gene targeting. Both male and female animals survive to adulthood with normal gross external phenotypes. Females are infertile; males have a decreased fertility. Females have hypoplastic uteri and hyperemic ovaries with no detectable corpora lutea. In adult wild-type and heterozygous females, 3-day estradiol treatment at 40 micrograms/kg stimulates a 3- to 4-fold increase in uterine wet weight and alters vaginal cornification, but the uteri and vagina do not respond in the animals with the estrogen receptor gene disruption. Prenatal male and female reproductive tract development can therefore occur in the absence of estradiol receptor-mediated responsiveness. PMID- 8248224 TI - Cytoplasmic dynein is required for normal nuclear segregation in yeast. AB - We have identified the gene DYN1, which encodes the heavy chain of cytoplasmic dynein in the yeast Saccharomyces cerevisiae. The predicted amino acid sequence (M(r) 471,305) reveals the presence of four P-loop motifs, as in all dyneins known so far, and has 28% overall identity to the dynein heavy chain of Dictyostelium [Koonce, M. P., Grissom, P. M. & McIntosh, J. R. (1992) J. Cell Biol. 119, 1597-1604] with 40% identity in the putative motor domain. Disruption of DYN1 causes misalignment of the spindle relative to the bud neck during cell division and results in abnormal distribution of the dividing nuclei between the mother cell and the bud. Cytoplasmic dynein, by generating force along cytoplasmic microtubules, may play an important role in the proper alignment of the mitotic spindle in yeast. PMID- 8248225 TI - Altered phosphorylation of growth-associated protein B50/GAP-43 in Alzheimer disease with high neurofibrillary tangle density. AB - The growth-associated phosphoprotein B50/GAP-43, associated with axonal proliferation and regeneration, was isolated from superior temporal gyrus (area 22) of seven control and eight Alzheimer disease (AD) postmortem human brains. Membrane and cytoplasmic proteins were fractionated and B50/GAP-43 was isolated by reverse-phase HPLC and gel electrophoresis. B50/GAP-43 was identified with rabbit polyclonal antibodies 4P3 (generated against the calmodulin binding domain of B50/GAP-43) and 1B5 (generated against whole bovine B50/GAP-43). B50/GAP-43 protein was further separated into phosphorylated and dephosphorylated species by calmodulin-Sepharose chromatography. The amounts of phosphorylated and dephosphorylated B50/GAP-43 forms were determined by electrophoresis, protein staining, and densitometry. Data on the relative phosphorylation of B50/GAP-43 protein in membrane and cytoplasmic fractions show a 10-fold difference in the ratio of cytoplasmic/membrane phosphorylation of B50/GAP-43 in AD brains with high neurofibrillary tangle (NFT) density compared to AD brains with low NFT density. This difference is due to a decreased percentage of phosphorylated B50/GAP-43 in the membrane fraction relative to that in the cytosolic fraction from high NFT density. No analogous relationship was found between the phosphorylation of B50/GAP-43 and the density of neuritic plaques in the brains examined. These data indicate differential distribution of phosphorylated and dephosphorylated B50/GAP-43 in normal and AD brains is related to NFT density but not to neuritic plaque density. PMID- 8248226 TI - Inhibition of protein synthesis alters the subcellular distribution of mRNA in neurons but does not prevent dendritic transport of RNA. AB - This study evaluates whether protein synthesis plays a role in targeting RNA molecules to different subcellular domains within neurons. Transport of newly synthesized RNA (labeled with [3H]uridine) was examined in the presence of the protein synthesis inhibitors puromycin and cycloheximide. In situ hybridization was used to determine whether inhibition of protein synthesis altered the subcellular distribution of mRNAs. Transport of recently synthesized RNA was not disrupted after prolonged exposure to either inhibitor. However, inhibition of protein synthesis caused several mRNAs that are normally confined to the cell body to appear in dendrites. The distribution of mRNAs that are normally present in dendrites was unaffected. These findings suggest that protein synthesis is not required to translocate RNA into the dendrites but may play a role in restricting particular mRNAs to the neuronal cell body. PMID- 8248227 TI - Two motion systems with common and separate pathways for color and luminance. AB - We present psychological experiments that reveal two motion systems, a specific and an unspecific one. The specific system prevails at medium to high temporal frequencies. It comprises at least two separate motion pathways that are selective for color and for luminance and that do not interact until after the motion signal is extracted separately in each. By contrast, the unspecific system prevails at low temporal frequencies and it combines color and luminance signals at an earlier stage, before motion extraction. The successful implementation of an efficient and accurate technique for assessing equiluminance corroborates further the main findings. These results offer a general framework for understanding the nature of interactions between color and luminance signals in motion perception and suggest that previously proposed dichotomies in motion processing may be encompassed by the specific/unspecific dichotomy proposed here. PMID- 8248228 TI - Newly expressed progesterone receptor cannot activate stable, replicated mouse mammary tumor virus templates but acquires transactivation potential upon continuous expression. AB - During development and differentiation, the expression of transcription factors is regulated in a temporal fashion. Newly expressed transcription factors must interact productively with target genes organized in chromatin. Although the mechanisms governing factor binding to chromatin templates are not well understood, it is now clear that template access can be dramatically influenced by nucleoprotein structure. We have examined the ability of a well characterized transactivator, the progesterone receptor (PR), to activate the mouse mammary tumor virus (MMTV) promoter organized either in stable, replicating templates that have a highly ordered nucleosome structure or as transiently transfected DNA, which adopts a less-defined structure. If the PR is transiently expressed in cells harboring both replicated and transient MMTV receptor constructs, it cannot significantly activate the stable replicated MMTV template. In contrast, when PR cDNA is stably inserted into the same cells and constitutively expressed, it gains the ability to activate both chromosomal and transiently introduced templates. These results demonstrate that newly expressed PR is not competent to activate the MMTV template in its native nucleoprotein conformation but acquires this ability upon prolonged expression in replicating cells. PMID- 8248229 TI - Use of yeast artificial chromosomes (YACs) for studying control of gene expression: correct regulation of the genes of a human beta-globin locus YAC following transfer to mouse erythroleukemia cell lines. AB - We demonstrate that transfer of a yeast artificial chromosome (YAC) containing 230 kb of the human beta-globin locus into mouse erythroleukemia cells by fusion results in correct developmental regulation of the human beta-like globin genes. Additionally, we show that early after hybrid formation, human embryonic epsilon- and fetal gamma-globin genes are coexpressed with the adult beta gene but that after 10-20 weeks in culture, globin gene expression switches to predominantly adult. Thus, in contrast to shorter gene constructs, the globin genes of the beta globin locus YAC are regulated like the chromosomal globin genes. These results indicate that transfer of YACs into established cell lines can be used for the analysis of the developmental control of multigenic and developmentally regulated human loci. PMID- 8248230 TI - Transgenic sorghum plants via microprojectile bombardment. AB - Transgenic sorghum plants have been obtained after microprojectile bombardment of immature zygotic embryos of a drought-resistant sorghum cultivar, P898012. DNA delivery parameters were optimized based on transient expression of R and C1 maize anthocyanin regulatory elements in scutellar cells. The protocol for obtaining transgenic plants consists of the delivery of the bar gene to immature zygotic embryos and the imposition of bialaphos selection pressure at various stages during culture, from induction of somatic embryogenesis to rooting of regenerated plantlets. One in about every 350 embryos produced embryogenic tissues that survived bialaphos treatment; six transformed callus lines were obtained from three of the eight sorghum cultivars used in this research. Transgenic (T0) plants were obtained from cultivar P898012 (two independent transformation events). The presence of the bar and uidA genes in the T0 plants was confirmed by Southern blot analysis of genomic DNA. Phosphinothricin acetyltransferase activity was detected in extracts of the T0 plants. These plants were resistant to local application of the herbicide Ignite/Basta, and the resistance was inherited in T1 plants as a single dominant locus. PMID- 8248231 TI - Simian virus 40 large tumor antigen is unable to transform mouse embryonic fibroblasts lacking type 1 insulin-like growth factor receptor. AB - Fibroblast cell lines were established from mouse embryos homozygous for a targeted disruption of the Igf1r gene, encoding the type 1 receptor for insulin like growth factor I (IGF-I) and from their wild-type littermates. The cells from the wild-type embryos (W cells) grow in serum-free medium supplemented with platelet-derived growth factor, epidermal growth factor, and IGF-I, whereas the cells from Igf1r(-/-) embryos (R- cells) do not, although they grow at a reduced rate in 10% fetal calf serum. The simian virus 40 (SV40) large T antigen, expressed from a transfected plasmid, can transform W cells, which form foci in monolayer cultures and colonies in soft agar (anchorage-independent growth). In contrast, the SV40 large tumor antigen, although normally expressed from the transfected template, is unable to transform R- cells, which remain contact inhibited and fail to grow in soft agar. The transformed phenotype is restored if the R- cells carrying the SV40 large tumor antigen are stably transfected with a plasmid expressing the human IGF-I receptor. These results demonstrate that signaling via the IGF-I receptor is an indispensable component of the SV40 transformation pathway. This conclusion is further supported from the results of antisense RNA experiments with tumor cell lines showing that interference with the function of the IGF-I receptor has a profound effect on anchorage-independent growth, even under conditions that only modestly affect growth in monolayers. PMID- 8248232 TI - Origin of the human alcohol dehydrogenase system: implications from the structure and properties of the octopus protein. AB - In contrast to the multiplicity of alcohol dehydrogenase in vertebrates, a class III type of the enzyme [i.e., a glutathione-dependent formaldehyde dehydrogenase; formaldehyde; NAD+ oxidoreductase (glutathione-formylating), EC 1.2.1.1.] is the only form detectable in appreciable yield in octopus. It is enzymatically and structurally highly similar to the human class III enzyme, with limited overall residue differences (26%) and only a few conservative residue exchanges at the substrate and coenzyme pockets, reflecting "constant" characteristics of this class over wide time periods. It is distinct from the ethanol-active "variable" class I type of the enzyme (i.e., classical liver alcohol dehydrogenase; alcohol:NAD+ oxidoreductase, EC 1.1.1.1). The residue conservation of class III is also spaced differently from that of class I but is typical of that of proteins in general, emphasizing that class I, with divergence at three functional segments, is the form with deviating properties. In spite of the conservation in class III, surface charges differ considerably. The apparent absence of a class I enzyme in octopus and the constant nature of the class III enzyme support the concept of a duplicative origin of the class I line from the ancient class III form. Still more distant relationships define further enzyme lines that have subunits with other properties. PMID- 8248233 TI - Identifying the catalytic residue of the ATPase reaction of DNA gyrase. AB - We propose a mechanism for the hydrolysis of ATP by the DNA gyrase B protein in which Glu42 acts as a general base and His38 has a role in aligning and polarizing the glutamate residue. We have tested this mechanism by site-directed mutagenesis, converting Glu42 to Ala, Asp, and Gln, and His38 to Ala. In the presence of wild-type A protein, B proteins bearing the mutations Ala42 and Gln42 show no detectable supercoiling or ATPase activities, while Asp42 and Ala38 proteins have reduced activities. In the DNA cleavage and relaxation reactions of gyrase, which do not require ATP hydrolysis, wild-type and mutant proteins have similar activities. When the 43-kDa N-terminal fragment of the gyrase B protein (which hydrolyzes ATP) contained the mutations Ala42 or Gln42, ATP was bound but not hydrolyzed, supporting the idea that Glu42 is involved in hydrolysis but not nucleotide binding. PMID- 8248234 TI - Cloning and characterization of a transmembrane serine kinase that acts as an activin type I receptor. AB - Activin type II receptors are transmembrane protein-serine/threonine kinases. By using a reverse-transcription PCR assay to screen for protein kinase sequences, we isolated a cDNA clone, activin X1 receptor, from rat brain that encodes a 55 kDa transmembrane protein-serine kinase which is structurally related to other receptors in this kinase subfamily. The predicted protein consists of 509 amino acids, and the kinase domain shows 40% and 37% identity to the activin and transforming growth factor beta type II receptors, respectively. No activin binding was observed when activin X1 receptor was expressed alone in COS-M6 cells; however, coexpression with type II activin receptors gave rise to a 68-kDa affinity-labeled complex in addition to the 85-kDa type II receptor complex. The size of this cross-linked band is consistent with the size of the type I activin receptor; furthermore, activin X1 receptor associated with type II receptors, as judged by coimmunoprecipitation with type II receptor antibodies. These data suggest that activin X1 receptor can serve as an activin type I receptor and that the diverse biological effects of activins may be mediated by a complex formed by the interaction of two transmembrane protein-serine kinases. PMID- 8248235 TI - A human mitochondrial ATP-dependent protease that is highly homologous to bacterial Lon protease. AB - We have cloned a human ATP-dependent protease that is highly homologous to members of the bacterial Lon protease family. The cloned gene encodes a protein of 963 amino acids with a calculated molecular mass of 106 kDa, slightly higher than that observed by Western blotting the protein from human tissues and cell lines (100 kDa). A single species of mRNA was found for this Lon protease in all human tissues examined. The protease is encoded in the nucleus, and the amino terminal portion of the protein sequence contains a potential mitochondrial targeting presequence. Immunofluorescence microscopy suggested a predominantly mitochondrial localization for the Lon protease in cultured human cells. A truncated LON gene, in which translation was initiated at Met118 of the coding sequence, was expressed in Escherichia coli and produced a protease that degraded alpha-casein in vitro in an ATP-dependent manner and had other properties similar to E. coli Lon protease. PMID- 8248236 TI - Evidence for myosin motors on organelles in squid axoplasm. AB - Squid axoplasm has proved a rich source for the identification of motors involved in organelle transport. Recently, squid axoplasmic organelles have been shown to move on invisible tracks that are sensitive to cytochalasin, suggesting that these tracks are actin filaments. Here, an assay is described that permits observation of organelles moving on unipolar actin bundles. This assay is used to demonstrate that axoplasmic organelles move on actin filaments in the barbed-end direction, suggesting the presence of a myosin motor on axoplasmic organelles. Indeed, axoplasm contains actin-dependent ATPase activity, and a pan-myosin antibody recognized at least four bands in Western blots of axoplasm. An approximately 235-kDa band copurified in sucrose gradients with KI-extracted axoplasmic organelles, and the myosin antibody stained the organelle surfaces by immunogold electron microscopy. The myosin is present on the surface of at least some axoplasmic organelles and thus may be involved in their transport through the axoplasm, their movement through the cortical actin in the synapse, or some other aspect of axonal function. PMID- 8248237 TI - Evolution of an enzyme activity: crystallographic structure at 2-A resolution of cephalosporinase from the ampC gene of Enterobacter cloacae P99 and comparison with a class A penicillinase. AB - The structure of the class C ampC beta-lactamase (cephalosporinase) from Enterobacter cloacae strain P99 has been established by x-ray crystallography to 2-A resolution and compared to a class A beta-lactamase (penicillinase) structure. The binding site for beta-lactam (penicillinase) structure. The binding site for beta-lactam antibiotics is generally more open than that in penicillinases, in agreement with the ability of the class C beta-lactamases to better bind third-generation cephalosporins. Four corresponding catalytic residues (Ser-64/70, Lys-67/73, Lys-315/234, and Tyr-150/Ser-130 in class C/A) lie in equivalent positions within 0.4 A. Significant differences in positions and accessibilities of Arg-349/244 may explain the inability of clavulanate-type inhibitors to effectively inactivate the class C beta-lactamases. Glu-166, required for deacylation of the beta-lactamoyl intermediate in class A penicillinases, has no counterpart in this cephalosporinase; the nearest candidate, Asp-217, is 10 A from the reactive Ser-64. A comparison of overall tertiary folding shows that the cephalosporinase, more than the penicillinase, is broadly similar to the ancestral beta-lactam-inhibited enzymes of bacterial cell wall synthesis. On this basis, it is proposed that the cephalosporinase is the older of the two beta-lactamases, and, therefore, that a local refolding in the active site, rather than a simple point mutation, was required for the primordial class C beta-lactamase to evolve to the class A beta-lactamase having an improved ability to catalyze the deacylation step of beta-lactam hydrolysis. PMID- 8248238 TI - Analysis of the human alpha-globin gene cluster in transgenic mice. AB - A 350-bp segment of DNA associated with an erythroid-specific DNase I hypersensitive site (HS-40), upstream of the alpha-globin gene cluster, has been identified as the major tissue-specific regulator of the alpha-globin genes. However, this element does not direct copy number-dependent or developmentally stable expression of the human genes in transgenic mice. To determine whether additional upstream hypersensitive sites could provide more complete regulation of alpha gene expression we have studied 17 lines of transgenic mice bearing various DNA fragments containing HSs -33, -10, -8, and -4, in addition to HS -40. Position-independent, high-level expression of the human zeta- and alpha-globin genes was consistently observed in embryonic erythroid cells. However, the additional HSs did not confer copy-number dependence, alter the level of expression, or prevent the variable down-regulation of expression in adults. These results suggest that the region upstream of the human alpha-globin genes is not equivalent to that upstream of the beta locus and that although the two clusters are coordinately expressed, there may be differences in their regulation. PMID- 8248239 TI - A receptor for interleukin 10 is related to interferon receptors. AB - We isolated cDNAs encoding a mouse interleukin 10 receptor (mIL-10R) from mouse mast cell and macrophage cell lines. The two cDNAs are substantially identical and express an approximately 110-kDa polypeptide in COS7 cells, which binds mIL 10 specifically. A mouse pro-B-cell line (Ba/F3) expressing transfected recombinant mIL-10R binds IL-10 with high affinity (approximately 70 pM) and proliferates in response to mIL-10. mIL-10R is structurally related to interferon receptors (IFNRs). Since IL-10 inhibits macrophage activation by IFN-gamma, a possible implication of this relationship interaction of IL-10R and IFN-gamma R or their signaling pathways. PMID- 8248240 TI - A ubiquitous protein is the source of naturally occurring peptides that are recognized by a CD8+ T-cell clone. AB - We previously isolated from mouse spleen an octapeptide (LSPFPFDL) that in association with the class I major histocompatibility complex protein Ld is recognized by the antigen-specific receptor of an alloreactive CD8+ T-cell clone (2C). Guided by an assay dependent upon the same 2C T-cell receptor, we have now isolated from the same source another naturally occurring peptide. The second peptide (VAITRIEQLSPFPFDL) includes the entire octapeptide sequence and preliminary evidence suggests that it may be a natural precursor of the octapeptide. On finding extensive sequence homology between the 16-mer and part of human 2-oxoglutarate dehydrogenase, we determined the cDNA sequence of mouse 2 oxoglutarate dehydrogenase and found that the deduced amino acid sequence matches precisely the two naturally occurring peptides, indicating their origin by cellular processing of this ubiquitous self protein. PMID- 8248241 TI - Regulation by the neuropeptide cholecystokinin (CCK-8S) of protein phosphorylation in the neostriatum. AB - Despite physiological evidence that cholecystokinin (CCK) is an excitatory neurotransmitter in the brain, little is known about its mechanism of action. CCK immunoreactivity in the brain, including projections to the striatum, is primarily attributable to the sulfated octapeptide CCK-8S. We report here that CCK-8S abolishes cAMP-dependent phosphorylation of a dopamine- and cAMP-regulated 32-kDa phosphoprotein (DARPP-32) in striatal neurons. The effect of CCK-8S is prevented by antagonists of CCKB and N-methyl-D-aspartate receptors. Our results support a model in which CCK-8S, originating from CCK or CCK/glutamate corticostriatal neurons, promotes the release of an excitatory neurotransmitter that causes the dephosphorylation and inactivation of DARPP-32, a potent protein phosphatase inhibitor, thereby modulating neuronal excitability. PMID- 8248242 TI - Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease. AB - A neuropathological hallmark of Alzheimer disease (AD) is a widespread amyloid deposition. We analyzed the entire amino acid sequences in an amyloid preparation and found, in addition to the major beta/A4-protein (A beta) fragment, two unknown peptides. We raised antibodies against synthetic peptides using subsequences of these peptides. These antibodies immunostained amyloid in neuritic and diffuse plaques as well as vascular amyloid. Electron microscopic analysis demonstrated that the immunostaining was localized on amyloid fibrils. We have isolated an apparently full-length cDNA encoding a 140-amino-acid protein within which two previously unreported amyloid sequences are encoded in tandem in the most hydrophobic domain. We tentatively named this 35-amino acid peptide NAC (non-A beta component of AD amyloid) and its precursor NACP. NAC is the second component, after A beta, identified chemically in the purified AD amyloid preparation. Secondary structure predictions indicate that the NAC peptide sequence has a strong tendency to form beta-structures consistent with its association with amyloid. NACP is detected as a M(r) 19,000 protein in the cytosolic fraction of brain homogenates and comigrates on immunoblots with NACP synthesized in Escherichia coli from NACP cDNA. NACP mRNA is expressed principally in brain but is also expressed in low concentrations in all tissues examined except in liver, suggesting its ubiquitous and brain-specific functions. The availability of the cDNA encoding full-length NACP should help to elucidate the mechanisms of amyloidosis in AD. PMID- 8248243 TI - Importance of independence in ribozyme reactions: kinetic behavior of trimmed and of simply connected multiple ribozymes with potential activity against human immunodeficiency virus. AB - The kinetic behavior of ribozymes derived from two types of multiple-ribozyme expression vector were examined. In some cases, multiple ribozymes were expressed as a single RNA molecule and all the ribozymes were simply connected in tandem (connected type). In other cases, multiple ribozymes were flanked by cis-acting ribozymes at both their 5' and 3' ends so that, upon transcription, multiple ribozymes were trimmed at both their 5' and 3' ends, with resultant liberation of multiple independent ribozymes (shotgun type). When levels of ribozyme expression were examined for the shotgun-type vector, the level of the ribozyme transcript was found to be proportional to the number of units (n) connected in tandem. Accordingly, the activities of the shotgun-type ribozymes, in terms of the cleavage of HIV-1 RNA in vitro, were also found to be proportional to the number of units connected in tandem (n). By contrast, the activities of the connected type ribozymes reached plateau values at around n = 3. These results indicate that, when the shotgun-type expression system is used, it is theoretically possible to generate various independent ribozymes, each specific for a different target site, without sacrificing the activity of any individual ribozyme. PMID- 8248244 TI - Direct gene transfer with DNA-liposome complexes in melanoma: expression, biologic activity, and lack of toxicity in humans. AB - Direct gene transfer offers the potential to introduce DNA encoding therapeutic proteins to treat human disease. Previously, gene transfer in humans has been achieved by a cell-mediated ex vivo approach in which cells from the blood or tissue of patients are genetically modified in the laboratory and subsequently returned to the patient. To determine the feasibility and safety of directly transferring genes into humans, a clinical study was performed. The gene encoding a foreign major histocompatibility complex protein, HLA-B7, was introduced into HLA-B7-negative patients with advanced melanoma by injection of DNA-liposome complexes in an effort to demonstrate gene transfer, document recombinant gene expression, and determine the safety and potential toxicity of this therapy. Six courses of treatment were completed without complications in five HLA-B7-negative patients with stage IV melanoma. Plasmid DNA was detected within biopsies of treated tumor nodules 3-7 days after injection but was not found in the serum at any time by using the polymerase chain reaction. Recombinant HLA-B7 protein was demonstrated in tumor biopsy tissue in all five patients by immunochemistry, and immune responses to HLA-B7 and autologous tumors could be detected. No antibodies to DNA were detected in any patient. One patient demonstrated regression of injected nodules on two independent treatments, which was accompanied by regression at distant sites. These studies demonstrate the feasibility, safety, and therapeutic potential of direct gene transfer in humans. PMID- 8248245 TI - Tissue-specific expression of the human aromatase cytochrome P-450 gene by alternative use of multiple exons 1 and promoters, and switching of tissue specific exons 1 in carcinogenesis. AB - Extensive screening of aromatase cDNA was carried out in cDNA libraries from various human tissues. The DNA sequences of all the isolated cDNA clones were identical in the region encoded by exons 2-10 of the aromatase gene. However, tissue-specific sequences, which were classified into four groups, were observed in the 5' portions of the clones corresponding to the region encoded by exon 1. All of them were also found in clones isolated from a human genomic library and mapped between exons 1 and 2 of the human aromatase gene reported previously, suggesting the presence of multiple exons 1 and promoters in the gene. Reverse transcription-PCR analyses of aromatase mRNAs in various tissues revealed that aromatase transcripts are tissue-specifically spliced by alternative use of multiple exons 1, although minor forms of the transcripts were also present in each tissue. Aromatase mRNA is spliced from 10 exons in most tissues, but from 9 exons in the prostate and from 10 or 11 exons in the placenta. This suggests that tissue-specific regulation of the aromatase gene in various tissues may be explained by alternative use of multiple exons 1 flanked with tissue-specific promoters. The alternative use of multiple exons 1 for liver transcripts was found to change developmentally. Furthermore, switch from an adipose-specific exon 1 to another type of exon 1 was observed in aromatase transcripts of adipose tissues of three of five breast cancer patients. PMID- 8248246 TI - Structure and promoter characterization of the gene encoding the large subunit (R1 protein) of mouse ribonucleotide reductase. AB - Mammalian ribonucleotide reductase (EC 1.17.4.1) is composed of two nonidentical subunits, proteins R1 and R2, both required for enzyme activity. The structure of the genomic mouse ribonucleotide reductase R1 gene was compiled from a number of overlapping lambda clones isolated from a Charon 4A mouse sperm genomic library. The R1-encoding gene covers 26 kb and consists of 19 exons. All exon-intron boundaries were located by dideoxynucleotide sequencing, showing that intron 7 starts with the variant GC instead of GT. About 3.5 kb of DNA from the 5' flanking region of the R1-encoding gene were cloned and sequenced, and the transcriptional start site was determined by nuclease S1 mapping of RNA. DNase I footprinting assays on the R1 promoter identified two nearly identical 23-bp-long protein-binding regions. Three protein complexes binding to one of the 23-mer regions were resolved and partially identified by using gel-retardation mobility shift assays and UV crosslinking. One complex most likely contained Sp1, and another complex showed S-phase-specific binding, suggesting a direct role in the cell-cycle-dependent R1 gene expression. PMID- 8248247 TI - A peptide motif that recognizes A.T tracts in DNA. AB - The DAT1 gene of Saccharomyces cerevisiae encodes a DNA binding protein that specifically interacts with nonalternating oligo(A).oligo(T) tracts (A.T tracts). Deletion analysis of DAT1 coding information showed that the amino-terminal 36 residues are sufficient for specific DNA binding activity. Furthermore, a 35 residue synthetic peptide corresponding to amino acids 2-36 bound to A.T tracts with an equilibrium dissociation constant of 4 x 10(-10) M. Within this region the pentad Gly-Arg-Lys-Pro-Gly is repeated three times. Mutational analysis revealed that the Arg side chains are required for high-affinity binding, whereas the other pentad side chains are dispensable. Chemical interference experiments showed that the DAT1 protein interacts with the minor groove of the double helix. The data suggest that the pentad arginines interact in a cooperative manner with a repeated minor groove feature of A.T tract DNA to achieve high-affinity recognition. Amino acid similarities with other DNA binding proteins suggest that the DAT1 protein pentad represents a specialized example of a widespread motif used by proteins to recognize A.T base pairs. PMID- 8248248 TI - Helix capping propensities in peptides parallel those in proteins. AB - Helix content of peptides with various uncharged nonaromatic amino acids at either the N-terminal or C-terminal position has been determined. The choice of N terminal amino acid has a major effect on helix stability: asparagine is the best, glycine is very good, and glutamine is the worst helix-stabilizing amino acid at this position. The rank order of helix stabilization parallels the frequencies of these amino acids at the N-terminal boundary (N-cap) position of helices in proteins found by Richardson and Richardson [Richardson, J. S. & Richardson, D. C. (1988) Science 240, 1648-1652], and the N-terminal amino acid in a peptide composed of helix-forming amino acids may be considered as the N-cap residue. The choice of C-terminal amino acid has only a minor effect on helix stability. N-capping interactions may be responsible for the asymmetric distribution of helix content within a given peptide found by various workers. An acetyl group on the N-terminal alpha-amino function cancels the N-cap effect and the acetyl group is equivalent to N-terminal asparagine in an unacetylated peptide. Our results demonstrate a close relationship between the mechanisms of alpha-helix formation in peptides and in proteins. PMID- 8248249 TI - Charged histidine affects alpha-helix stability at all positions in the helix by interacting with the backbone charges. AB - To determine whether a charged histidine side chain affects alpha-helix stability only when histidine is close to one end of the helix or also when it is in the central region, we substitute a single histidine residue at many positions in two reference peptides and measure helix stability and histidine pKa. The position of a charged histidine residue has a major effect on helix stability in 0.01 M NaCl: the helix content of a 17-residue peptide is 24% when histidine is at position 3 compared to 76% when it is at position 17. This dependence of helix content on histidine position decreases sharply in 1 M NaCl, as expected for counterion screening of the charge-helix dipole interaction. Results at interior positions indicate that the position of a charged histidine residue affects helix stability at these positions. Unexpectedly high values of the helix content are found when either neutral or charged histidine is at one of the last three C-terminal positions, suggesting that either form can stabilize an isolated helix by hydrogen bonding to a main-chain CO group. PMID- 8248250 TI - Elastic free energy of anisotropic helical ribbons as metastable intermediates in the crystallization of cholesterol. AB - We report measurements of the geometrical structure and temporal evolution of metastable helical intermediates in the pathway for cholesterol crystallization in native and model biles. We find that the lecithin component in the bile can dramatically affect the kinetics along this pathway. We also present a theoretical description of these helical intermediates using an elastic free energy appropriate for anisotropic bilayers of tilted chiral amphiphiles, which provides a quantitative description of the observed helical ribbon geometry and insight into the relative free energies of the observed metastable intermediates. PMID- 8248252 TI - Erythropoietin receptor signals both proliferation and erythroid-specific differentiation. AB - Ectopic expression of the erythropoietin receptor (EPO-R) in Ba/F3, an interleukin 3-dependent progenitor cell line, confers EPO-dependent cell growth. To examine whether the introduced EPO-R could affect differentiation, we isolated Ba/F3-EPO-R subclones in interleukin 3 and assayed for the induction of beta globin mRNA synthesis after exposure to EPO. Detection of beta-globin mRNA was observed within 3 days of EPO treatment, with peak levels accumulating after 10 days. When EPO was withdrawn, expression of beta-globin mRNA persisted in most clones, suggesting that commitment to erythroid differentiation had occurred. Although EPO-R expression also supports EPO-dependent proliferation of CTLL-2, a mature T-cell line, those cells did not produce globin transcripts, presumably because they lack requisite cellular factors involved in erythrocyte differentiation. We conclude that the EPO-R transmits signals important for both proliferation and differentiation along the erythroid lineage. PMID- 8248251 TI - Transgenic mice expressing the human GLUT4/muscle-fat facilitative glucose transporter protein exhibit efficient glycemic control. AB - To examine the physiological role of the GLUT4/muscle-fat specific facilitative glucose transporter in regulating glucose homeostasis, we have generated transgenic mice expressing high levels of this protein in an appropriate tissue specific manner. Examination of two independent founder lines demonstrated that high-level expression of GLUT4 protein resulted in a marked reduction of fasting glucose levels (approximately 70 mg/dl) compared to wild-type mice (approximately 130 mg/dl). Surprisingly, 30 min following an oral glucose challenge the GLUT4 transgenic mice had only a slight elevation in plasma glucose levels (approximately 90 mg/dl), whereas wild-type mice displayed a typical 2- to 3-fold increase (approximately 250-300 mg/dl). In parallel to the changes in plasma glucose, insulin levels were approximately 2-fold lower in the transgenic mice compared to the wild-type mice. Furthermore, isolated adipocytes from the GLUT4 transgenic mice had increased basal glucose uptake and subcellular fractionation indicated elevated levels of cell surface-associated GLUT4 protein. Consistent with these results, in situ immunocytochemical localization of GLUT4 protein in adipocytes and cardiac myocytes indicated a marked increase in plasma membrane associated GLUT4 protein in the basal state. Taken together these data demonstrate that increased expression of the human GLUT4 gene in vivo results in a constitutively high level of cell surface GLUT4 protein expression and more efficient metabolic control over fluctuations in plasma glucose concentrations. PMID- 8248253 TI - Identification of the promoter and a transcriptional enhancer of the gene encoding L-CAM, a calcium-dependent cell adhesion molecule. AB - L-CAM is a calcium-dependent cell adhesion molecule that is expressed in a characteristic place-dependent pattern during development. Previous studies of ectopic expression of the chicken L-CAM gene under the control of heterologous promoters in transgenic mice suggested that cis-acting sequences controlling the spatiotemporal expression patterns of L-CAM were present within the gene itself. We have now examined the L-CAM gene for sequences that control its expression and have found an enhancer within the second intron of the gene. A 2.5-kb Kpn I-EcoRI fragment from the intron acted as an enhancer of a simian virus 40 minimal promoter driving a chloramphenicol acetyltransferase (CAT) reporter gene and produced 14.0-fold induction of CAT activity in MDCK cells. To narrow down the region responsible for enhancer activity and to determine whether the enhancer could function in a cell type-specific manner, a number of smaller restriction fragments from the intron were tested for activity in two chicken cell lines, the LMH hepatoma line, which produces high levels of L-CAM, and the SL-29 fibroblast line, which produces little, if any, L-CAM. Four L-CAM enhancer plasmids containing shorter segments derived from the intron showed enhanced CAT activity levels (between 9.4- and 16.5-fold) in extracts from transfected LMH cells but not from SL-29 cells. DNA sequence analysis of the L-CAM enhancer region revealed putative binding sites for the transcription factors SP1, E2A, and AP-2. In addition, LE-9, the smallest L-CAM enhancer segment (310 bp), contained a consensus binding site for the liver-enriched POU-homeodomain transcription factor, HNF-1. Tests of upstream sequences showed that a 630-bp fragment, corresponding to nearly the entire intergenic region between L-CAM and its neighboring CAM gene, K-CAM, could function as a promoter. In combination with the L-CAM enhancer, this fragment directed cell type-specific expression of the CAT reporter gene in LMH cells at a level comparable to that observed with enhancer constructs using the simian virus 40 minimal promoter. These combined observations define a promoter and an enhancer for the chicken L-CAM gene. They raise the possibility that these cis-acting regulatory sequences may be instrumental in directing specific place-dependent expression of the L-CAM gene in the chicken. PMID- 8248254 TI - Selective repression of transcriptional activators at a distance by the Drosophila Kruppel protein. AB - The Kruppel (Kr) protein, bound at kilobase distances from the start site of transcription, represses transcription by RNA polymerase II in mammalian cells. Repression is monotonically dependent on the dose of Kr protein and the presence of Kr binding site(s) on the DNA. These data suggest an inhibitory protein protein interaction between the Kr protein and proximal transcription factors. Repression by Kr depends on the specific activator protein driving transcription. In particular, Kr protein selectively represses transcription mediated by the Sp1 glutamine-rich activation domain, tethered to the promoter by a GAL4 DNA-binding domain, but does not repress transcription stimulated by the acidic GAL4 activator. We believe this represents repression by a quenching interaction between DNA-bound Kr protein and the activation region of Sp1, rather than competition between Sp1 and Kr for a limiting transcriptional component. Selective, context-related repression affords an added layer of combinatorial control of gene expression by sequence-specific transcription factors. PMID- 8248255 TI - Isolation of cDNA encoding the human NF-E2 protein. AB - The human homolog of mouse NF-E2 was isolated from the K562 cell line and found to encode a member of the basic leucine-zipper family of DNA-binding regulatory proteins. The deduced amino acid sequence of the mouse and human proteins exhibited near identity. Comparison to the related protein, Nrf1, revealed significant homologies at isolated regions, particularly within the basic domain, suggesting that NF-E2 and Nrf1 are members of a distinct subfamily of basic leucine-zipper proteins that share similar DNA-binding properties. High levels of human NF-E2 mRNA were observed in human erythroleukemic cell lines examined. Extensive survey of human tissue samples found NF-E2 expression not limited to erythropoeitic organs. Expression in the colon and testis suggests that NF-E2 may participate in the regulation of genes other than globin. PMID- 8248256 TI - Cloning of Nrf1, an NF-E2-related transcription factor, by genetic selection in yeast. AB - We have devised a complementation assay in yeast to clone mammalian transcriptional activators and have used it to identify a human basic leucine zipper transcription factor that we have designated Nrf1 for NF-E2-related factor 1. Nrf1 potentially encodes a 742-aa protein and displays marked homology to the mouse and human NF-E2 transcription factors. Nrf1 activates transcription via NF E2 binding sites in yeast cells. The ubiquitous expression pattern of Nrf1 and the range of promoters containing the NF-E2 binding motif suggest that this gene may play a role in the regulation of heme synthesis and ferritin genes. PMID- 8248257 TI - The enhancer of position-effect variegation of Drosophila, E(var)3-93D, codes for a chromatin protein containing a conserved domain common to several transcriptional regulators. AB - In Drosophila modifying mutations of position-effect variegation have been successfully used to genetically dissect chromatin components. The enhancer of position-effect variegation E(var)3-93D [formerly E-var(3)3] encodes proteins containing a domain common to the transcriptional regulators tramtrack and the products of the Broad complex. It interacts with a number of chromatin genes that suppress position-effect variegation. Mutations in E(var)3-93D exhibit an imprinting-like effect on the Y chromosome. This effect is transmitted paternally over several generations. Homeotic transformations in E(var)3-93D mutants indicate an involvement of the gene products in regulation of homeotic gene complexes. An antiserum raised against E(var)3-93D protein detects this chromosomal protein in a large subset of sites in polytene chromosomes. Our genetic and molecular data suggest that the proteins of E(var)3-93D are generally involved in establishing and/or maintaining an open chromatin conformation. PMID- 8248258 TI - Germ-line transmission and developmental regulation of a 150-kb yeast artificial chromosome containing the human beta-globin locus in transgenic mice. AB - Sequential expression of the genes of the human beta-globin locus requires the formation of an erythroid-specific chromatin domain spanning > 200 kb. Regulation of this gene family involves both local interactions with proximal cis-acting sequences and long-range interactions with control elements upstream of the locus. To make it possible to analyze the interactions of cis-acting sequences of the human beta-globin locus in their normal spatial and sequence context, we characterized two yeast artificial chromosomes (YACs) 150 and 230 kb in size, containing the entire beta-globin locus. We have now successfully integrated the 150-kb YAC into the germ line of transgenic mice as a single unrearranged fragment that includes the locus control region, structural genes, and 30 kb of 3' flanking sequences present in the native locus. Expression of the transgenic human beta-globin locus is tissue- and developmental stage-specific and closely follows the pattern of expression of the endogenous mouse beta-globin locus. By using homology-directed recombination in yeast and methods for the purification and transfer of YACs into transgenic mice, it will now be feasible to study the physiological role of cis-acting sequences in specifying an erythroid-specific chromatin domain and directing expression of beta-globin genes during ontogeny. PMID- 8248259 TI - The Drosophila melanogaster flightless-I gene involved in gastrulation and muscle degeneration encodes gelsolin-like and leucine-rich repeat domains and is conserved in Caenorhabditis elegans and humans. AB - Mutations at the flightless-I locus (fliI) of Drosophila melanogaster cause flightlessness or, when severe, incomplete cellularization during early embryogenesis, with subsequent abnormalities in mesoderm invagination and in gastrulation. After chromosome walking, deficiency mapping, and transgenic analysis, we have isolated and characterized flightless-I cDNAs, enabling prediction of the complete amino acid sequence of the 1256-residue protein. Data base searches revealed a homologous gene in Caenorhabditis elegans, and we have isolated and characterized corresponding cDNAs. By using the polymerase chain reaction with nested sets of degenerate oligonucleotide primers based on conserved regions of the C. elegans and D. melanogaster proteins, we have cloned a homologous human cDNA. The predicted C. elegans and human proteins are, respectively, 49% and 58% identical to the D. melanogaster protein. The predicted proteins have significant sequence similarity to the actin-binding protein gelsolin and related proteins and, in addition, have an N-terminal domain consisting of a repetitive amphipathic leucine-rich motif. This repeat is found in D. melanogaster, Saccharomyces cerevisiae, and mammalian proteins known to be involved in cell adhesion and in binding to other proteins. The structure of the maternally expressed flightless-I protein suggests that it may play a key role in embryonic cellularization by interacting with both the cytoskeleton and other cellular components. The presence of a highly conserved homologue in nematodes, flies, and humans is indicative of a fundamental role for this protein in many metazoans. PMID- 8248260 TI - Amino acid substitutions in the floor of the putative antigen-binding site of H 2T22 affect recognition by a gamma delta T-cell receptor. AB - We have previously identified a self-reactive gamma delta T-cell clone (KN6) specific for the H-2T region gene product T22b. Now we have investigated by an in vitro mutagenesis analysis of the T22b gene the possibility that the interaction between the KN6 gamma delta T-cell receptor and T22b involves a peptide. The results demonstrate that mutations at the floor of the putative antigen-binding groove of T22b affect recognition by the gamma delta T-cell receptor. Furthermore, we have shown that KN6 cells react with cells that are deficient in the class I peptide transporter TAP1/TAP2. These results suggest that peptide is involved in the interaction of the KN6 T-cell receptor with T22 and that loading of T22 with the putative peptide is TAP1/TAP2-independent. PMID- 8248261 TI - Restoration of early thymocyte differentiation in T-cell receptor beta-chain deficient mutant mice by transmembrane signaling through CD3 epsilon. AB - Thymic repertoire selection requires the expression of the alpha beta CD3 T-cell receptor (TCR) together with the coreceptors CD4 and CD8. The appearance of CD4 and CD8 on thymocytes is the hallmark of a complex maturation step, accompanied by downregulation of the interleukin 2 receptor (IL-2R) alpha chain, arrest of rearrangement (i.e., allelic exclusion) of the TCR beta-chain locus, a burst of cell divisions, and reduction in cell size. This maturation step is inhibited in TCR beta-chain-deficient mouse strains and may depend on surface expression of an immature TCR complex containing CD3 and TCR beta chains but no TCR alpha chain. Here we show that the CD4+8+ double-positive (DP) stage can be induced by treatment of fetal thymic organ cultures with anti-CD3 epsilon monoclonal antibodies in several TCR beta-chain-deficient mouse strains: severe combined immunodeficient (scid) mice, mice carrying a mutation in the recombination activating gene 1 (Rag-1), or mice carrying a deletion in the TCR beta-chain locus itself. These findings suggest that CD3 epsilon is expressed on the thymocyte surface independent of and prior to the TCR beta chain. The data are consistent with the notion that in wild-type mice the DP stage is induced by transmembrane signaling through an immature CD3-TCR beta-chain complex, which can be bypassed by crosslinking of CD3 epsilon alone. PMID- 8248262 TI - Human cytomegalovirus induces JC virus DNA replication in human fibroblasts. AB - JC virus, a human papovavirus, is the causative agent of the demyelinating brain disease progressive multifocal leucoencephalopathy (PML). PML is a rare but fatal disease which develops as a complication of severe immunosuppression. Latent JC virus is harbored by many asymptomatic carriers and is transiently reactivated from the latent state upon immunosuppression. JC virus has a very restricted host range, with human glial cells being the only tissue in which it can replicate at reasonable efficiency. Evidence that latent human cytomegalovirus is harbored in the kidney similar to latent JC virus led to the speculation that during episodes of impaired immunocompetence, cytomegalovirus might serve as helper virus for JC virus replication in otherwise nonpermissive cells. We show here that cytomegalovirus infection indeed leads to considerable JC virus DNA replication in cultured human fibroblasts that are nonpermissive for the replication of JC virus alone. Cytomegalovirus-mediated JC virus replication is dependent on the JC virus origin of replication and T antigen. Ganciclovir-induced inhibition of cytomegalovirus replication is associated with a concomitant inhibition of JC virus replication. These results suggest that reactivation of cytomegalovirus during episodes of immunosuppression might lead to activation of latent JC virus, which would enhance the probability of subsequent PML development. Ganciclovir induced repression of both cytomegalovirus and JC virus replication may form the rational basis for the development of an approach toward treatment or prevention of PML. PMID- 8248263 TI - Induction of long-term facilitation in Aplysia sensory neurons by local application of serotonin to remote synapses. AB - Long-term synaptic facilitation at the connections of Aplysia sensory neurons onto their target cells involves alterations in gene expression. How then are the relevant cellular signals for the induction and expression of long-term synaptic changes conveyed between the nucleus and remote synaptic terminals? We have explored this question using a set of remote, peripheral terminals of siphon sensory cells, which are approximately 3 cm from the sensory cell body in the abdominal ganglion. We found that these remote synapses, like the proximal synapses previously studied in dissociated cell culture, can exhibit long-term facilitation 24 hr after cell-wide serotonin application. Furthermore, serotonin applications restricted to the remote synaptic terminals nevertheless produced long-term facilitation, indicating that signals generated in synaptic regions can trigger the long-term process, perhaps via retrograde signals to the nucleus to modify gene expression, followed by anterograde signals back to the terminal. Serotonin applications restricted to the cell body and proximal synapses of the sensory neuron also produced long-term facilitation at remote synapses, although to a lesser extent, suggesting that long-term facilitation is expressed cell wide, but that superimposed on this cell-wide facilitation there appears to be a component that is synapse-specific. PMID- 8248264 TI - Isolation of Hsp90 mutants by screening for decreased steroid receptor function. AB - The 90-kDa heat shock protein Hsp90 represents a highly conserved strongly expressed gene family; in Saccharomyces cerevisiae, Hsp90 proteins are essential for cell viability. Hsp90 interacts with certain cellular proteins, including steroid hormone receptors, tyrosine and serine/threonine kinases, and other heat shock proteins, but its biological functions are not understood. The unliganded glucocorticoid receptor must interact with Hsp90 to acquire competence for high affinity hormone binding and subsequent transcriptional regulation. By screening in yeast for defects in glucocorticoid receptor function, Hsp90 mutants were isolated. Four such mutants are described, all of which interact with the glucocorticoid receptor but display distinct defects in ligand responsiveness and differences in growth and resistance to high temperature. PMID- 8248265 TI - The Arabidopsis endoplasmic reticulum retention receptor functions in yeast. AB - Soluble proteins retained in the lumen of the endoplasmic reticulum (ER) contain a carboxyl-terminal tetrapeptide sequence that functions presumably to recycle these proteins from a subsequent compartment. Biochemical and genetic evidence indicate that the ERD2 gene product is the receptor for these ER retention signals. Here we report the identification of a cDNA clone from Arabidopsis thaliana (aERD2) similar in sequence and size to members of the ERD2 gene family. Southern and Northern blot analyses indicate that Arabidopsis contains a single aERD2 gene which is expressed at different levels in various plant tissues. A functional assay demonstrates that the Arabidopsis homologue, unlike the mammalian protein, can complement the lethal phenotype of the erd2 deletion mutant of Saccharomyces cerevisiae, indicating that this protein may have a similar function in plants. As the plant protein may have a binding specificity similar to the human Erd2 protein but can function in yeast, we suggest that the plant homologue is the functional link between yeast and animals. PMID- 8248266 TI - Interrelationship between dietary protein, cholesterol and n-6 polyunsaturated fatty acid metabolism. PMID- 8248267 TI - The synthesis of rare and unusual fatty acids. PMID- 8248269 TI - Monthly Prostaglandin Bibliography prepared by the University of Sheffield Biomedical Information Service. PMID- 8248268 TI - Transport of lipids to the plasma membrane in animal cells. PMID- 8248270 TI - Effects of fish oils on lipid variables and platelet function indices. PMID- 8248271 TI - Dietary sunflower, linseed and fish oils affect phospholipid fatty acid composition, development of cardiac lesions, phospholipase activity and eicosanoid production in Atlantic salmon (Salmo salar). AB - Atlantic salmon (Salmo salar) post-smolts were fed practical-type diets in which the lipid was supplied either as fish oil (FO), sunflower oil (SFO) or linseed oil (LO) for 12 weeks. In general, the heart phospholipids from SFO-fed fish had increased 18:2n-6, 20:2n-6, 20:3n-6 and 20:4n-6 but decreased 20:5n-3 compared to both other dietary treatments. This was reflected in a decreased n-3/n-6 polyunsaturated fatty acid (PUFA) ratio and an increased 20:4n-6/20:5n-3 or eicosanoid precursor ratio in SFO-fed fish. While heart phospholipids of fish fed LO had increased levels of 18:2n-6, 20:2n-6 and 20:3n-6 compared to fish fed FO, 20:4n-6 levels were reduced, although only significantly in phosphatidylcholine (PC). Dietary-induced changes in phospholipid fatty acid compositions of blood leucocytes were similar to those in heart, although fish fed LO had increased 20:5n-3 compared to fish fed FO. Thromboxane B2 (TXB2) produced by stimulated blood cells was reduced in fish fed LO compared to those fed SFO. Prostaglandin E2 (PGE2) production was reduced in LO-fed fish compared to both other dietary treatments. Fish fed LO had reduced PC in heart membranes compared to the other two dietary treatments, resulting in a ratio of PC:PE (phosphatidylethanolamine) less than unity. Fish fed SFO developed a marked cardiac histopathology which, while present in FO-fed fish albeit in a less severe form, was virtually absent in fish fed LO. Fish fed SFO had increased heart phospholipase A activity compared to those given either FO or LO. PMID- 8248272 TI - Mononuclear cell-conditioned medium enhances thrombin-stimulated PGI2 production by human umbilical vein endothelial cells in culture. AB - We investigated the effect of blood mononuclear cell-conditioned medium on prostacyclin (PGI2) production by human umbilical vein endothelial cells in culture (HUVEC), and compared the potency of the conditioned medium in PGI2 production with that of various cytokines and lipopolysaccharide (LPS). HUVEC which had been preincubated with LPS, interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor (TNF alpha), or interferon-gamma (IFN-gamma) produced more PGI2 than control cells in response to thrombin. However, the HUVEC preincubated with the conditioned medium made with mononuclear cells with or without LPS (LPS-Mo-CM, Mo-CM) produced more PGI2 than those preincubated with LPS, IL-1 alpha, IL-1 beta, TNF alpha, or IFN-gamma. Although the concentrations or IL-1 beta and TNF alpha in the post-culture medium of HUVEC treated with LPS Mo-CM were much higher than those with Mo-CM, LPS-Mo-CM which was made with 13,000/ml of mononuclear cells and 1 microgram/ml of LPS did not significantly augment the subsequent PGI2 production by HUVEC as compared with Mo-CM made with the same numbers of mononuclear cells. PGI2 production by Mo-CM-treated HUVEC still exceeded that of control cells, even when an excess amount of antibody to TNF alpha and/or IL-1 alpha was added to the Mo-CM. It is possible that Mo-CM contains unknown cytokines besides IL-1 and TNF which stimulate the HUVEC to produce PGI2. PMID- 8248273 TI - The influence of indomethacin on the hatching of mouse blastocysts. AB - In order to investigate the effect of indomethacin for inhibition of the cyclooxygenase pathway on the hatching process of mouse blastocysts, 508 mouse blastocysts were cultured in modified HAM F10 medium containing 0, 8, 79 or 788 microM indomethacin, added after 24 h of incubation. Hatching was scored after 72 h of incubation. In another series of experiments, indomethacin was added only after blastulation had occurred. Indomethacin in doses of 8 microM and 79 microM, added after 24 h of incubation or after blastulation had occurred, did not influence successful hatching. A dose of 788 microM indomethacin interfered with hatching, but this dose appeared to be toxic for the embryo. PMID- 8248274 TI - Pilot trial to determine the efficacy of a low dose of fish oil in the treatment of angina pectoris in the geriatric patient. AB - 23 geriatric patients completed a single-blind placebo controlled, crossover trial using fish oil as adjunctive therapy for stable angina pectoris. The results indicated a significant reduction in the number of anginal attacks, as well as a significant reduction in the consumption of sublingual isosorbide dinitrate tablets. A possible prolonged carry over effect of fish oil was noted after crossover from the active substance to placebo. PMID- 8248275 TI - Dietary supplementation with primrose oil or fish oil does not change urinary excretion of prostacyclin and thromboxane metabolites in pre-eclamptic women. AB - To study whether balance between antiaggregatory, vasodilatory prostacyclin (PGI2) and proaggregatory, vasoconstrictory thromboxane A2 (TXA2) could be affected by dietary manipulation, 18 pre-eclamptic women were treated in randomized order between 31 and 36 weeks of gestation either with primrose oil (n = 7), with fish oil (n = 5), or with placebo (n = 6). Urinary excretions of the degradation products of PGI2 (6-keto-PGF1 alpha, 2,3-dinor-6-keto-PGF1 alpha) and TXA2 (TXB2, 2,3-dinor-TXB2) were measured in 24 h urines before and serially during the supplementation. Fatty acid supplementation did not affect urinary prostanoid excretions or clinical signs of pre-eclampsia. PMID- 8248276 TI - Effect of policosanol on cerebral ischemia in Mongolian gerbils: role of prostacyclin and thromboxane A2. AB - Policosanol is a mixture of higher primary aliphatic alcohols, isolated from sugar cane wax, whose main component is octacosanol. Policosanol (25, 50 and 200 mg/kg) administered by the oral route not only significantly reduced serum thromboxane B2 (TXB2) levels but also, at 200 mg/kg significantly increased 6 keto-PGF1 alpha in Mongolian gerbils. Policosanol at 200 mg/kg significantly protected against cerebral ischemia induced by unilateral ligation of common carotid artery in Mongolian gerbils. In this experimental model, combined administration of ineffective doses of policosanol (25 mg/kg) and aspirin (ASA) (30 mg/kg) significantly protected animals indicating a synergism between them. PMID- 8248277 TI - Prostacyclin release in experimental diabetes: effects of evening primrose oil. AB - Alterations in release of endothelium-derived vasomotor agents could underlie microvascular and neuropathic complications in diabetes. This study examined release of the potent vasodilator prostacyclin, measured as immunoreactive 6-keto prostaglandin F1 alpha, from rat lung, kidney and peripheral nerve. Tissues were taken from control and streptozotocin-diabetic rats which had been treated for 8 weeks with either evening primrose oil (EPO) or, as a control for lipid intake, coconut oil (CO). Lung and kidney slices were incubated in the presence of acetylcholine (ACh), the calcium ionophore 4-Br-A23187, arachidonic acid (AA) or without agonist (basal). Segments of sciatic nerve, with their epineuria punctured, were incubated with or without 4-Br-A23187. Basal prostacyclin release from the lung was significantly higher in rats treated with EPO irrespective of diabetic state (increased by 60% in controls and by 77% in diabetics). Levels were reduced in CO-diabetics compared to EPO-controls (53% reduction) and CO controls (30% reduction), although this did not reach statistical significance in the latter. Basal prostacyclin release was also significantly reduced in the kidney from CO-diabetics (40% reduction compared to CO-controls and 56% reduction compared to EPO-controls). In the presence of AA, lung prostacyclin release was significantly lower in CO-diabetic rats compared to all other groups (40% reduction compared to EPO-diabetics and 60% compared to both control groups) but there were no differences in renal release between any group. Prostacyclin release by nerves from CO-diabetic rats was significantly reduced (by 91-93%) compared to all other groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248278 TI - Enhancement of antineoplastic effect and attenuation of sister chromatid exchanges by prostaglandin E2 in Ehrlich ascites tumour cells treated with cyclophosphamide in vivo. AB - Reduced sister chromatid exchanges (SCE) frequency in response to cyclophosphamide (CP) was observed when Ehrlich ascites tumour (EAT) cells were exposed in vivo to 2 micrograms/g body weight of prostaglandin E2 (PGE2). 1 h before i.p. injection of 5-bromodeoxyuridine (BrdUrd) adsorbed to activated charcoal, EAT-bearing mice treated i.p. with CP appeared to have increased SCE rates and cell division delays. PGE2 had no effect on survival and in inhibiting tumour growth. CP had only a slight non-significant effect on survival and in inhibiting tumour growth. In mice treated with the combined CP (5 micrograms/g bd wt) plus PGE2 (2 micrograms/g bd wt) a significant enhancement (P < 0.01) of survival time was accompanied by inhibition of tumour growth (P < 0.01) in comparison with the untreated controls. These data imply that SCEs might result from errors in a repair process which might involve a PGE2 sensitive step. PMID- 8248279 TI - Modulation of prostaglandin E2-induced Ca2+ influx by steroid hormones in osteoblast-like cells. AB - In osteoblast-like MC3T3-E1 cells, we previously reported that prostaglandin E2 (PGE2), a potent bone resorbing agent, stimulates Ca2+ influx (H. Tokuda, M. Miwa, Y. Oiso and O. Kozawa, Cell Signal 1992; 4: 261-266). In this study, we examined the effects of various hormones belonging to the steroid hormone superfamily on PGE2-induced Ca2+ influx in MC3T-E1 cells. 1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3], an active form of vitamin D3, dexamethasone and retinoic acid significantly inhibited the PGE2-induced Ca2+ influx in a dose-dependent manner in these cells. The effects of these hormones were dependent on the time of pretreatment and submaximum inhibitions were observed at 6 h. In contrast, 24,25 dihydroxyvitamin D3 [24,25-(OH)2D3], an inactive form of vitamin D3, 17 beta estradiol, progesterone, testosterone and triiodothyronine had little effect on the PGE2-induced Ca2+ influx in these cells. These results suggest that, in the steroid hormone superfamily, 1,25-dihydroxyvitamin D3, glucocorticoid and retinoic acid modulate bone metabolism through the inhibition of Ca2+ influx induced by PGE2 in osteoblast-like cells. PMID- 8248280 TI - Peptidoleukotrienes increase the efflux of glutathione from perfused rat liver. AB - The effect of peptidoleukotrienes (LT) on the efflux of glutathione (GSH) from the perfused rat liver was investigated. LTD4, C4 and E4 were infused at a final concentration of 20 nM into the portal vein of rat livers perfused with Krebs Henseleit buffer. Perfusion pressure, efflux of glucose and release of LDH increased during the infusion of LTC4 and D4 and returned to baseline upon cessation of the infusion of LT. In contrast, the efflux of GSH did not change during the infusion of LT, but increased from 15 +/- 2 to 26 +/- 4 nmol/min.g liver 20 min after cessation of the infusion of LTC4. LTE4 did not increase the efflux of LDH, glucose, lactate, or GSH. During the LTC4- and LTD4-induced rise in perfusion pressure bile-flow decreased transiently by one third. The biliary excretion of GSH, however, decreased by an average of 75% and recovered more slowly than the cholestasis. In the presence of the selective LTD4 receptor antagonist LY171883 the effects of LTC4 and LTD4 were largely abolished. The delayed effects of LT on GSH efflux suggest that LT shift the efflux of GSH from the canalicular towards the sinusoidal side of the hepatocyte independent of other effects of LT on hepatic function. The sustained increase in efflux of GSH resulting from LT will raise the extracellular concentration of this antioxidant, such that more GSH is available at sites of inflammation to detoxify reactive oxygen species released by activated inflammatory cells. PMID- 8248281 TI - Pharmacology of lower urinary tract smooth muscles and penile erectile tissues. PMID- 8248282 TI - Neuropharmacology of quinolinic and kynurenic acids. AB - In a little more than 10 years, the kynurenine metabolites of tryptophan have emerged from their former position as biochemical curiosities, to occupy a prominent position in research on the causes and treatment of several major CNS disorders. The pathway includes two compounds, quinolinic acid and kynurenic acid, which are remarkably specific in their pharmacological profiles: one is a selective agonist at receptors sensitive to NMDA, whereas the other is a selective antagonist at low concentrations at the strychnine-resistant glycine modulatory site associated with the NMDA receptor. It has been argued that these agents cannot be of physiological or pathological relevance because their normal extracellular concentrations, in the nanomolar range, are at least 3 orders of magnitude lower than those required to act at NMDA receptors. This is a facile argument, however, that ignores at least two possibilities. One is that both quinolinate and kynurenate may be present in very high concentrations locally at some sites in the brain that cannot be reflected in mean extracellular levels. Similar considerations apply to many neuroactive agents in the CNS. The fact that both compounds appear to be synthesised in, and thus emerge from, glial cells that are well recognised as enjoying a close physical and chemical relationship with some neurones in which the intercellular space may be severely restricted may support such a view. Certainly the realisation that NMDA receptors may not be fully saturated functionally with glycine would be consistent with the possibility that even quite low concentrations of kynurenate could maintain a partial antagonism at the glycine receptor. A second possibility is that there may be a subpopulation of NMDA receptors (or, indeed, for a quite different amino acid) that possesses a glycine modulatory site with a much lower sensitivity to glycine or higher sensitivity to kynurenate, making it more susceptible to fluctuations of endogenous kynurenine levels. Whatever the specific nature of their physiological roles, the presence of an endogenous selective agonist and antagonist acting at NMDA receptors must continue to present exciting possibilities for understanding the pathological basis of several CNS disorders as well as developing new therapeutic approaches. An imbalance in the production or removal of either of these substances would be expected to have profound implications for brain function, especially if that imbalance were present chronically.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8248283 TI - Axon guidance factors in invertebrate development. AB - The contribution that studies in the invertebrates have made to our understanding of the factors responsible for directing axon growth is reviewed. Cellular mechanisms for axon guidance are considered, particularly the question of the accuracy of initial axon growth, and the implications of these observations for models of growth cone turning. The cellular substrates followed by growing axons during embryogenesis are identified, together with the experimental evidence that each is essential for reliable axon navigation. The significance of these studies for investigations into the molecular nature of axon guidance factors is discussed, and the likely cellular roles of putative axon guidance molecules considered. PMID- 8248284 TI - Regulation of steroid hydroxylase gene expression: importance to physiology and disease. AB - Steroid hydroxylase gene expression is multifactorial in nature, being regulated by tissue-specific, developmental, constitutive and signal transduction systems. The biochemistry of this complex pattern of regulation is not yet clearly elucidated, but studies in several laboratories have led to an understanding of specific aspects of regulation, particularly that involving signal transduction. The complexity of regulation appears to be necessary for normal human physiology because of the wide variety of steroid hormones produced by these enzymes. Genetic diseases associated with the steroid hydroxylases provide examples of how aberrant physiology can result from alterations in the multifactorial regulation of steroid hydroxylase gene expression. PMID- 8248285 TI - The role of atrial natriuretic peptide (ANP) in chronic liver disease. AB - The role of atrial natriuretic peptide (ANP) and potential defects of ANP in liver disease are reviewed. Patients with cirrhosis of the liver show no decrease of ANP plasma concentrations nor changes in the pattern of ANP immunoreactivity nor changes of splanchnic ANP clearance. The renal effects of exogenously administered as well as endogenously released ANP are blunted in cirrhosis, in particular in patients with ascites. This seems due to increased activity of sodium-retaining hormonal systems and changes of the renal ANP receptor status. Pharmacological inhibition of ANP-degradation or clearance may yield therapeutic potential. PMID- 8248286 TI - Current developments in proton therapy: a review. AB - The use of high-energy protons in radiotherapy was first proposed in 1946. In the last decade there has been a significant growth in the number of centres using protons in the treatment of malignant and non-malignant disease. To date (January 1993) a total of more than 11,500 patients have been treated world-wide. Encouraging clinical results have been reported in the literature. The purpose of this article is to outline the advantages of proton beams and to review current developments in physics and engineering applied to the field of proton therapy with particular emphasis on proton accelerator technology and the development of proton therapy facilities. The production of clinically useful beams is discussed and the relative merits of different treatment systems compared. Reference is also made to the factors affecting the absorbed dose in a patient and to proton radiobiology together with the results of studies of comparisons of treatment planning with protons with that using conventional photon therapy. The dosimetry of proton beams is also reviewed. PMID- 8248287 TI - Determining locations of intracerebral lesions for proton radiotherapy. AB - Using an external view of the head, the ability to direct a proton radiotherapy beam at an intracerebral lesion is described, using a biostereometric (BSM) technique. The positions of markers, recognizable on computed tomography (CT) or magnetic resonance (MR) images, at known locations on the skin of a patient's head are related to the site of the lesion seen on the images. The markers' positions are determined by a BSM device, incorporating six video cameras coupled to a computer. A transformation from a coordinate system, defined by the CT or MR images, to another coordinate system, that of the BSM device, allows the proton beam to be directed towards the lesion. Markers used in this work were solid steel spheres for CT and plastic spheres containing oil for MR images. PMID- 8248288 TI - Modelling photon transport in non-uniform media for SPECT with a vectorized Monte Carlo code. AB - A vectorized Monte Carlo code has been developed for modelling photon transport in non-uniform media for single-photon-emission computed tomography (SPECT). The code is designed to compute photon detection kernels, which are used to build system matrices for simulating SPECT projection data acquisition and for use in matrix-based image reconstruction. Non-uniform attenuating and scattering regions are constructed from simple three-dimensional geometric shapes, in which the density and mass attenuation coefficients are individually specified. On a Stellar GS1000 computer, Monte Carlo simulations are performed between 1.6 and 2.0 times faster when the vector processor is utilized than when computations are performed in scalar mode. Projection data acquired with a clinical SPECT gamma camera for a line source in a non-uniform thorax phantom are well modelled by Monte Carlo simulations. The vectorized Monte Carlo code was used to stimulate a 99Tcm SPECT myocardial perfusion study, and compensations for non-uniform attenuation and the detection of scattered photons improve activity estimation. The speed increase due to vectorization makes Monte Carlo simulation more attractive as a tool for modelling photon transport in non-uniform media for SPECT. PMID- 8248289 TI - Quality assurance in brachytherapy: principles for ionization chamber measurement of absorbed dose close to brachytherapy sources. AB - At present there exists no standardized procedure for absorbed dose determination with ionization chambers in the vicinity of brachytherapy sources. In this paper, a modification of the IAEA formalism for external photon beams is proposed in order to make it applicable at a reference point 20 mm from brachytherapy sources as well. A non-uniformity correction factor is introduced to correct for the effects of the steep absorbed dose gradient, and it is concluded that reasonable accuracy is possible when the Bragg-Gray principle is assumed to be valid. PMID- 8248291 TI - Special series: skeletal muscle, part 1. PMID- 8248290 TI - Quality assurance in brachytherapy: the displacement effect in the vicinity of 60Co and 192Ir brachytherapy sources. AB - Cylindrical ionization chambers with varying radii have been used to determine the displacement effect at distances between 10 mm and 40 mm from 60Co and 192Ir brachytherapy sources. Agreement with published values of the displacement factor for external 60Co beams is obtained. The shift between the centre of the ionization chamber and the effective point of measurement is found to be only a few per cent of the internal radius of the cylindrical ionization chamber. The shift is found to be dependent on the source to ionization chamber centre distance. PMID- 8248292 TI - Motor unit recruitment and the gradation of muscle force. AB - The capabilities of the different types of motor units are reviewed, and their properties in a variety of muscles are discussed. Because the tension-generating capacities of motor units are so different, the order in which they are recruited will have a strong influence on the way force output of the whole muscle is graded. Activation of motor units in a random order produces a roughly linear force increase with progressive recruitment, whereas recruitment of motor units in order of increasing force produces an approximately exponential force increase as the number of active motor units increases. The latter scheme allows fine control of weak movements and rapid production of powerful movements. Motor units are shown to be well adapted to the tasks they must perform, and a "compromise" motor unit will not fulfill all the tasks demanded of it. Finally, changes in motor unit properties produced by different activity patterns and by muscle reinnervation are reviewed, and the implications for rehabilitation are discussed. PMID- 8248293 TI - Skeletal muscle mechanics: implications for rehabilitation. AB - Skeletal muscles are the primary organ system responsible for force generation and movement. As such, an improved understanding of normal movement can be obtained by understanding skeletal muscle mechanical properties. In this review, we present the basic mechanical properties of skeletal muscle in a way that relates to their normal function. First, isometric force production is discussed, followed by a presentation of isotonic force production. Then, skeletal muscle architectural properties are presented as a strategy for muscles to specialize in either force production or excursion. Finally, we discuss the relationship between muscles and joints and the significance of this relationship for understanding strength. Based on this presentation, the therapist will have an improved understanding of normal movement and may have insights into developing rehabilitation protocols that can improve function. PMID- 8248294 TI - Compartmentalization of muscles and their motor nuclei: the partitioning hypothesis. AB - This review article is designed to expose physical therapists to an examination of muscle organization and the implications that this organization has for therapeutic applications. The partitioning hypothesis is based on the fact that an individual muscle is arranged in a more complex array than simply fibers attaching at aponeuroses, tendons, or bones with a single muscle nerve innervation. Neuromuscular compartments, which are distinct subvolumes of a muscle, each innervated by an individual muscle nerve branch and each containing motor unit territories with a unique array of physiological attributes, are described. In addition, the organization of individual muscles into these subunits is paralleled by the organization of their parent motoneurons within the spinal cord. These notions are detailed in a review of data derived from studies performed primarily in cat and rat models. Recent data derived from morphological and anatomical study of human muscles support the existence of similar neuromuscular partitions. These data are complemented by physiological studies, the results from which suggest that partitions may have functional or task oriented roles; that is, different portions of one muscle may be called into play depending on the task demands of the situation. The importance of these observations for reconsidering how we provide clinical applications, such as neuromuscular stimulation or kinesiological monitoring, is discussed. PMID- 8248295 TI - Motor unit populations in healthy and diseased muscles. AB - The numbers of functioning motor units can be estimated in proximal and distal muscles of human limbs by an electrophysiological technique in which the mean sizes of the motor unit potentials are compared with the maximum M-waves of the same muscles. Although manual methods of estimation have been used successfully in the past, the introduction of automated techniques has brought considerable advantages, including greater objectivity and reduced contamination of the results by "alternation." In healthy subjects, the intrinsic muscles of the hand have approximately 100 motor units each, and the biceps brachii muscle has only slightly more. With advancing age, there is a loss of motor units, which appears to be more pronounced in distal muscles. The motor unit estimating methodology has been found to be of value in the diagnosis and assessment of patients suspected of having muscle denervation. In amyotrophic lateral sclerosis, the mean rate of motor unit loss is swift, whereas in late-onset cases of spinal muscular atrophy, the reduction in the motor unit population does not appear to progress. In only the most rapidly deteriorating cases of post-polio syndrome is it possible to demonstrate further loss of motor units. In all of these denervating disorders, and in peripheral neuropathies, the importance of collateral reinnervation as a compensatory mechanism is emphasized. PMID- 8248296 TI - The role of skeletal muscle in glucose transport, glucose homeostasis, and insulin resistance: implications for physical therapy. AB - Skeletal muscle has a fundamentally important role in the maintenance of normal glucose homeostasis and in regulating whole-body carbohydrate metabolism. In this review, we discuss the regulation of skeletal muscle glucose transport by muscular activity and inactivity. A large number of patients routinely seen by physical therapists exhibit some form of skeletal muscle insulin resistance. Therefore, we discuss how skeletal muscle insulin resistance can be localized to a relatively small muscle mass, or in other circumstances can affect a large proportion of the muscle mass leading to disturbances in whole-body glucose homeostasis. We review the mechanisms and regulation of skeletal muscle glucose transport as background for understanding how defects in this process may contribute to the underlying pathogenesis of insulin resistance. Research into the events regulating glucose entry into skeletal muscles has considerable impact on how physical therapy exercise prescriptions may benefit patients with disturbances in carbohydrate metabolism. With an understanding of the principles of proper exercise prescription, physical therapists can use exercise training as a primary therapeutic intervention to improve local muscle and whole-body glucose utilization, and thereby minimize insulin resistance. PMID- 8248297 TI - Combined use of surface electromyography and 31P-NMR spectroscopy for the study of muscle disorders. AB - This article describes the use of combining spectral electromyographic signal techniques with phosphorus magnetic resonance (31P-NMR) spectroscopy for the purpose of studying muscle disorders. The quantification of muscle fatigue by electromyographic spectral variables such as the median frequency is summarized. Its development as a laboratory and clinical tool is presented, with an emphasis toward its potential as an assessment procedure. Similarly, the use of 31P-NMR spectroscopy for noninvasive measurement of phosphate metabolites and intracellular pH during fatigue are described. The limitations of this procedure are presented and compared with surface electromyographic techniques. Suggestions are made for combining these techniques for the purpose of monitoring muscle metabolic and electrophysiologic changes in situ during fatiguing exercises. A recent study in which these techniques were combined to evaluate the underlying mechanisms of fatigue in patients with fibromyalgia is described. PMID- 8248298 TI - Muscle fatigue: clinical implications for fatigue assessment and neuromuscular electrical stimulation. AB - Muscle fatigue can be defined as a decrease in the force-generating ability of a muscle that resulted from recent activity. Recent studies of muscle fatigue are reviewed that are relevant to two areas of interest to physical therapists: clinical assessment of muscle fatigue and neuromuscular electrical stimulation. Volitional and electrical tests have been used to quantify muscle fatigue. Several variations on each type of test are discussed, as are the possible sites in which fatigue might occur. The rate of fatigue during the therapeutic application of electrical stimulation of skeletal muscle is much greater than that seen during volitional contractions. Factors contributing to this phenomenon are examined. The unique requirements affecting how stimulus variables can be manipulated to minimize muscle fatigue in three specific therapeutic uses of neuromuscular electrical stimulation are addressed. PMID- 8248299 TI - Injury to skeletal muscle fibers during contractions: conditions of occurrence and prevention. AB - Contraction-induced injury results in the degeneration and regeneration of muscle fibers. Of the three types of contractions--shortening (concentric), isometric, and lengthening (eccentric)--injury is most likely to occur and the severity of the injury is greatest during lengthening contractions. The magnitude of the injury to muscle fibers may be assessed by direct measures of cellular and ultrastructural damage; by indirect measures of changes in enzyme efflux, calcium influx, ratio of oxidized to reduced glutathione, and force development; and, in human beings, by reports of muscle soreness. The sequence of events includes an initial injury that is primarily mechanical and a secondary metabolic, or biochemical, injury that peaks 1 to 3 days after the injurious contractions. The recovery from contraction-induced injury is usually complete within 30 days. Repeated exposures to protocols of lengthening contractions result in "trained" muscles that are not injured by the protocol that previously caused injury. PMID- 8248300 TI - Educational programs. PMID- 8248301 TI - [Venous disease, venous trauma and perspectives]. PMID- 8248302 TI - [Does the inferior vena cava have 2 surfaces and 2 borders also?]. AB - The vena cava that has a preferential flattening axis, has also 2 wall sides and 2 borders. The back wall fits closely round on the back vertebral plane and the main colaterals terminate on the borders of the vein. TM echography perfectly analyses the movements of the walls of the vein, as well as the respiratory and auricular movements. In a lying patient, echo-doppler colour shows the expiratory acceleration of the vena cava flow in subrenal area associated with an inspiratory slowing down (as well as for the femoral veins) and the inspiratory acceleration of the flow in suprarenal area associated with an expiratory slowing down. In a standing patient, the vena is cylindrical. When he/she walks on a treadmill, the diameter of the cava seems to be constant, i.e. quasi identical to the aorta's, as fluxes vary in the aorta according to the ventricular contractions and in the LVC according to the patient's gait. PMID- 8248303 TI - [Radiographic anatomy of the inferior azygos system, the ovarian and spermatic veins]. AB - The spermatic vein is tributary at the right of the infra-renal cava mainstem and at the left of the renal vein. It is fitted with an ostium or pre-ostium valvula which is normally tight. In the broad ligament of the uterus, the ovarian vein exchanges plexiform anastomosi with the homolateral or contralateral uterine vein through pre- or retro-uterine arches. The spermatic or ovarian reflux is more frequent on the left. Retrograde venography under cava occlusion is a good means to explore these vessels. The big Azygous vein joins the sub-renal vena cava by two roots. The internal root, which is often slender, perforates vertically the dialitic membrane. The external root, bigger, is the vein in L2; the lateral part of its track takes place in the psoas where it gains veins proceeding from the foramina of the adjacent conjugations. On the left side, the roots of the hemi Azygos appear most frequently in the left-renal vein. There may be a single root: the external root is the Lejars' arch. The variations of the derivative cavo-cava system represented by the Azygos veins are studied on the anatomical plan. Pathological literature complements this study. PMID- 8248304 TI - [Doppler pulsed and color echography of the inferior vena cava]. AB - B mode real time echotomography associated with duplex and colour doppler makes possible a complete investigation of the lower vena cava. The ilio-cava fossa is difficult to explore because of gas and digestive superpositions. Diagramatically, it is possible to distinguish five levels of investigation. 1) Iliac veins and ilio-cava convergence. 2) Sub-renal LVC. 3) Inter-renal LVC and communications with renal veins. 4) Retro-hepatic LVC: communications with liver and supra-hepatic veins. 5) Thoracic LVC and termination in the right atrium. Sector-based probes provide a 60 to 110 degrees sector thanks to which it is possible to obtain a sufficient exploration field for a very limited acoustic fenestra. Duplex and colour doppler assess the venous flow and its variations during the respiratory cycle and during the operations of compression or Valsalva's experiment. Colour doppler detects more precisely slow, collateral or repermeation flows. The recent complete thrombosis leads no duplex and colour doppler signal but an increase of the vein diameter associated with a collateralitis syndrome. In case of partial thrombosis, the vein is partially compressible as colour doppler fits thrombus closely round and visualizes the remaining lumen. In the ilio-cava fossa, compressions (either tumorous, ganglial or aneurysmal) which are often associated are diagnosed in the meanwhile. It is important to know the change of diameter and the abnormalities of the LVC position when a cava blocking is advised. A LVC whose diameters exceed 28 mm (mega-cava) contra indicated filter because of the risk of migration. Colour doppler makes easier the supervision of the blocking. PMID- 8248305 TI - [Imaging-anatomy correlation of the subrenal part of the inferior vena cava]. AB - A limited number of tomodensimetrical sections, MRI or echographical according to the fundamental, frontal, sagittal or horizontal plans makes possible the exploration of the sub-renal part of the lower vena cava. The comparison of pictures with the corresponding anatomical sections makes possible a detailed analysis of all the visualized structures, around the lower vena cava. PMID- 8248306 TI - [The ilio-caval confluence syndrome]. AB - The venous confluence syndrome is the clinical consequence of the flows obstruction which are the main tributary of the deep venous system. The cava confluence syndrome is different from the aortic obstruction at the level of its bifurcation. Its causes are congenital abnormality, extrinsic, intrinsic and intramural compression. CLINICAL PICTURE: collaterization of the int. vertebral plexus of the Azigos and hemiozygos veins as well as episgastric and thoracic veins, in chronic cases. On the other hand, in acute cases, significant renal insufficiency, ascites and bilateral phlegmasia caerulea make up the clinical picture. In acute stage, surgery requires thrombectomy and endovascular prothesis whereas in chronic stage, surgery will tend to remove the cause with only a palliative action on external symptoms. In both cases, abnormalities bring about a surgical issue. PMID- 8248307 TI - [Spinal canal stenosis syndrome by venous collateralization of an inferior cava thrombosis]. AB - We described in 1988 the case of a young patient with a right iliac vein and vena cava thrombosis who developed lubagoes and intermittent claudication of paralytic type. A digital venography by left femoral approach showed a derivation via the lumbar vein towards the spinal plexus. Later, the patient experienced a left external iliac vein thrombosis. Subsequently, collaterization went through the truncal veins and the clinical manifestations of the tight spinal canal disappeared. Since then, several cases of chronic thrombosis of the left primary iliac veins have been discovered. These also showed unexplained lumbagoes of the same duration. The idea of a relation of cause and effect was seldom accepted by the patients, which prevented us to proceed to many venous catherisms. Also, we performed only one operation until now: the cesarean of an ascending lumbar vein. In this very case, the tight spinal canal symptoms disappeared but appeared again after a few month-time for unknown reasons. Only one acute case has been found up to now: a woman who delivered by ligation suffered from lumbagoes and sciaticae. A dilatation of spinal veins and a lower vena cava thrombosis were showed by tomography. This woman had no leg symptom and lumbagoes disappeared with the partial recanalization of the cava. PMID- 8248308 TI - [Thrombophlebitis of the ovarian vein with a floating clot in the inferior vena cava]. AB - The authors report two recent observations of thrombophlebitis of the right ovarian vein. The first occurred after a cesarotomy, due to a bigeminal pregnancy, and the second, after a breech delivery. The diagnosis was given when faced to a febrile syndrome and pains of the right flank and confirmed in both cases by an abdominal tomodensimetric examination. In both cases, an enlargement of the lower vena cava thrombosis was observed with a floating clot that reached the renal veins. Both patients underwent a surgery. The latter consisted in both cases in a thrombectomy of the lower vena cava as well as a ligature of the right ovarian vein. In both cases, the evolution was positive, thanks to a remote control of the lower vena cava via tomodensimetry. This therapeutic procedure and other potential therapies were discussed. PMID- 8248309 TI - [In vitro bench tests of caval umbrella filters]. AB - Seven ombrelles percutaneous cava filters, now available, are tested on hydraulic testing ground. Measurements are concerned with the blocking function of filters and pressure changes induced in the cava flow and the cava track wall of the testing ground. A statistical analysis of the results (100 measurements per filter) shows homogeneous reactions of each filter when faced with different situations imposed by variations of the testing ground. Greenfield's filter, a reference, is compared with other models. This one and the L.G. filter show satisfactory filtering qualities. Two other filters, Cardial and Vascor, offer a highest filtering power. Other models give poor filtering results. PMID- 8248310 TI - [Blue phlebitis with exo- and endo-caval filters: 5 case reports]. AB - 5 cases of phlegmatia caerula dolens have been observed after the fixation of a cava blocking. You will find below the characteristics of the case reports: Mean age of patients: 69.2, from 55 to 83. Early phlegmatiae caeruleae dolens: 2 cases; late phlegmatiae: 3 cases (3 and 4 years after the cava ligature). Clinical context: advanced age; general state alteration 1 case; artery predisposition: 2 cases; heparin thrombopenia: 1 case. Responsible material: ombrelle de Mobin Uddin: 3 cases; Adams-De Weese's Clip: 2 cases. Current filters are probably less thrombogenous. Nevertheless, these case reports make us aware of the fact that in case of predisposition and/or in case of precary hemodynamic conditions, any factor likely to generate or worsen a venous stasis can originate (immediately or later) a significant thrombosis and, particularly in a few conditions, a phlegmatia caerulea dolens. Consequently, partial cava blocking indications must be seriously taken into consideration and saved for cases in which embolic risk is patent. PMID- 8248311 TI - [Percutaneous caval filter Dibie-Musset "DM". Results of animal experiments]. AB - AIM: a 7 F percutaneous cava filter was achieved, developed and tested in a goat. Thanks to its double-spiral original form, it is possible to place and remove it atraumaticaly and percutaneously. PRINCIPLES OF THE FILTER: its diameter, larger than the lower vena cava's (LVC) leads to a flattening of the venous lumen whose flow is intersected by the filter turns, thus creating a netting effect. CASE REPORT: during 28 months, 40 filters were introduced under fluorscopy in 20 goats. Thanks to 16 embolizations, it was possible to test the effectiveness of the filter against small emboli, with simultaneous cavography and pulmonary angiography in 4 cases. 29 filters were removed by jugular and femoral track, from D0 to D14. 12 goats were sacrificed to investigate histologicaly and macroscopicaly the LVC and control the biocompatibility from the 8th to the 385th day. Clinical and radiological supervision lasted more than one year for 3 goats. RESULTS: the size of the filter (30, 35, 40 mm) is chosen from the LVC diameter measured by cavography. 30 filters were introduced via jugular vein, 10 via femoral vein. These filters were introduced by catheter 7F thanks to an applicator and placed in correct position in the LVC. RELIABILITY OF THE FILTER: easy percutaneous introduction 7F. Once installed, the filters flatten the LVC; this process is automatically confirmed by cavography, and by scanning in 4 cases. EFFECTIVENESS AGAINST EMBOLI: out of 16 cases, 2 partial failures were observed at the beginning of the experiment (one spiral-fitted filter). Its effectiveness was optimized thanks to the addition of a second spiral which allowed the blocking of over-2 mm clots. PERCUTANEOUS REMOVAL: during the initial removals, partial failures were due to the fragility of the filter and the inflexibility of the extracting material. Successive changes of the shape and the alloy of the filter as well as the development of catheters and extracting materials have led to a sufficient reliability to remove (D0 to D14) the filter in security, by percutaneous tract (9 jugular, 9 femoral) before its clamping on the LVC, on the 15th day. INNOCUOUSNESS: both biological supervision and anatomo pathological investigation have showed the good tolerance of the filter. In local areas, this atraumatic filter does not wound nor perforate the LVC wall. Histologically, a thickening of the intima is observed. The positive results of this experiment led us to start clinical trials of "DM" filter in human beings. PMID- 8248312 TI - [Temporary caval filters. Our experience. Preliminary analysis of 24 cases]. AB - The AA. utilized temporary vena cava filters (16 Filcard and 8 Lysofilters) in 24 patients affected by deep venous thrombosis (DVT) of the lower limbs for the prevention of primary and recurrent pulmonary embolism (PE). The diagnosis of thromboembolic disease was always achieved by means of Ultrasounds (echo-color doppler) and was punctually confirmed by a retrograde cavagram during the insertion of the device. 19 patients presented large free-floating thrombi at inferior caval, iliac or common femoral vein level whereas 5 patients presented thrombi mostly of occlusive aspect. There was clinical or scintigraphic evidence of PE in 6 of the patients enrolled. 20 patients, without contraindications, were treated by fibrinolysis (F) with Urokinase (2-10 days) whereas 4 patients underwent surgical thrombectomy (T) because of short time relation with surgical intervention or trauma. All of them were protected by temporary vena cava filters and heparinized. All the filters were removed within 10 days. The results were considered "very good" (complete regression of floating thrombi) in 16 cases (14 F + 2 T), "good" (nearly complete regression of floating thrombi) in 3 cases (2 F + 1 T) and "poor" (unchanged) in the remaining 5 cases (4 F + 1 T). We didn't observe any new case or relapse of PE in the whole group and, furtherly, in 2 cases (1 F and 1 T) we demonstrated the capture of big emboli by the filter's basket. These clots were subsequently dissolved by fibrinolysis. To achieve the diagnosis of thromboembolic disease the following methods were used: 1- Screening: echo-color doppler of lower limbs extended to iliac and inferiora cava veins for detection of DVT and echocardio-color doppler for the detection of cardiac signs of PE. 2--DIAGNOSIS: pulmonary scintigram, retrograde cavogram and, rarely, angioCT scan. 3--FOLLOW-UP: echo-color doppler of lower limbs and pulmonary scintigram. The percutaneous insertion sites were the basilic vein (Filcard) and the right jugular vein (Lysofilter). Left jugular vein was used in 1 case with a big thyroid goitre. In the present experience we had no accidents during filters introduction or removal and no thrombosis at the insertion site (1 case of phlebitis of basilic vein). Indications and effectiveness: our results seem to be favorable to the use of inferior vena cava temporary filters for primary and recurrent pulmonary embolism prevention in the cases with floating thrombi both on fibrinolysis and embolectomy. In the cases of occlusive thrombotic diseases they proved to be effective to prevent PE during surgical embolectomy.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8248313 TI - [Preliminary results of the thrombolytic Angiocor in massive pulmonary emboli and proximal venous thrombosis]. PMID- 8248314 TI - [Prosthetic reconstructions of the inferior vena cava]. PMID- 8248315 TI - [Anomalies of the subrenal inferior vena cava in the surgery of non-specific and inflammatory abdominal aortic aneurysms]. AB - The sub-renal abnormalities of the lower vena cava (LVC) (left LVC, double LVC) are determined by a deterioration of the alteration process of supra-cardinal veins. Though they are rare, it is necessary to look for them during surgery of abdominal aorta in order to lower the risk of iatrogenic venous injuries. You will find below the description of six cases of sub-renal lower vena cava abnormality (3 double LVC, 3 left LVC) associated with an abdominal aorta aneurism (4 non specific aneurisms, 2 inflammations ones) as well as the diagnostic aspects and the technical issues they cause during the reconstruction of a non specific and inflammation aneurism of the abdominal aorta. PMID- 8248316 TI - [Resection of the aortic bifurcation for fibrosarcoma of the inferior vena cava]. AB - Leiomyosarcomas of the lower vena cava are tumours whose prognosis is bad. The best survival opportunities consist of the widest possible exeresis. You will find below two cases that lead us to proceed to a joint exeresis of the aortic and vena cava bifurcation. PMID- 8248317 TI - [Leiomyosarcoma of the great veins: a case involving the left iliac vein extending to the inferior vena cava]. AB - Large veins LMS is a rare slow growing malignant tumor originating from smooth muscle cells of the media. The authors report a case of LMS of the left common iliac vein propagating to the Inferior Vena Cava that presented with a left femoral-iliac deep thrombophlebitis. CT scan showed an uneven solid mass approximately 5 cm large within the left side of the pelvis. The mass displaced the left iliac artery and compressed the left iliac vein without a significant cleavage surface between the mass itself and the vascular structures. Location was next to the spine, medially and anteriorily to the psoas muscle. A thrombosis could be noticed within the distal segment of the inferior Vena Cava and within the proximal segment of the left iliac vein. US scan with fine needle biopsy of the mass didn't yield significant information. At surgical exploration a neoplastic mass involving and blocking the left iliac vein was found. Veinotomy performed on the iliac vein and on the distal segment of the Inferior Vena Cava but without infiltration of the vein walls. Surgical treatment consisted of asportation of the neoplastic mass, resection of the left iliac vein and thrombectomy of the Inferior Vena Cava. Histologic examination of the operated specimen revealed a mixoid LMS with vascular origin without involvement of the surrounding lymph nodes. Absence of clinical and radiological signs of relapse eight months after surgery makes further surgical and complementary (drug- and radiotherapy) treatments currently unnecessary. PMID- 8248318 TI - [All venous obstructions is not necessarily due to phlebitis. Case report of an ilio-femoral venous leiomyosarcoma]. AB - Venous leiomyosarcoma are rare malignant tumours that are usually revealed by an oedema of the lower limb, a sign of deep phlebitis. As the development is not linked with radiotherapy and chemotherapy, the prognosis depends mainly on a early diagnosis, which brings the best opportunities of complete surgical exeresis before the apparition of metastasis. The case reported below as well as ten other cases taken in the literature show the necessity to mention this diagnosis when confronted to any phlebitis appeared in the absence of usual precipitating factors. It is all the more necessary when mass is found by palpation, a small tumour is revealed by echography or when phlebography shows a lateral or circumferencial stricture that is different from usual pictures of phlebitis. PMID- 8248319 TI - [External valvuloplasty under preoperative angioscopic control]. AB - Intraoperative angioscopy was applied to evaluate the venous valvular incompetence and perform external valvuloplasty in case of venous reflux. Sixty seven limbs, 43 cases of primary varicose veins were examined using intraoperative angioscopy and the angioscopic findings of the incompetent venous valves were classified into three types as follows: valves with elongated and atrophic cusps 43 (50%)--type I, values with expanded and depressed commissures 36 (42%)--type II, and valves with perforated cusps or other changes 7 (8%)--type III, according to the angioscopie findings, external valvuloplasty was done in 31 subterminal valves of the long saphenous veins and 7 highest valves of the superficial femoral vein. Our external valvuloplasty consisted of two techniques. The first was the total plication technique for valvular annulus by a running suture of prolene and the second was by the venocuff sleeve of the autogenetic femorofascial band. The degree of plication was decided by angioscopic observation. Postoperative observation periods were from 2 to 28 months. There was no recurrence of varicose veins or prominent venous reflux. The application of intraoperative angioscopy is therefore useful for the choosing appropriate surgical procedures and for the evaluation of venous valvuloplasty. PMID- 8248320 TI - Photodegradation and in vitro phototoxicity of fenofibrate, a photosensitizing anti-hyperlipoproteinemic drug. AB - The phototoxic anti-hyperlipoproteinemic drug fenofibrate was found to be photolabile under aerobic and anaerobic conditions. Irradiation under argon of a methanol solution of this drug produced the photoproducts isopropyl 4-(1-[4 chlorophenyl]-1,2-dihydroxy)ethylphenoxyisobutyrate, 1,2-bis(4-chlorophenyl) 1,2bis (4-[isopropoxycarbonylisopropoxy]phenyl)ethane-1,2-diol and 4-(4 chlorobenzoyl)phenol, while under oxygen the photoproducts were 4 chloroperbenzoic acid, methyl 4-chlorobenzoate, 4-chlorobenzoic acid and singlet oxygen, as evidenced by trapping with 2,5-dimethylfuran. These results can be rationalized through hydrogen abstraction by excited fenofibrate, to afford a free radical as key intermediate. Biologically active antioxidants such as glutathione and cysteine efficiently reduced 4-chloroperbenzoic acid to 4 chlorobenzoic acid. The involvement of an electron transfer mechanism is suggested by detection (UV-vis spectrophotometry) of the radical cation TMP+. during the oxidation of tetramethylphenylenediamine (TMP) with 4-chloroperbenzoic acid. Fenofibrate was phototoxic in vitro when examined by the photohemolysis test, both under oxygen and argon atmosphere, although the photohemolysis rate was markedly lower under anaerobic conditions. The photoproducts 4-(1-[4 chlorophenyl]-1,2-dihydroxy)ethylphenoxyisobutyrate and 4-chloroperbenzoic acid induced hemolysis in the dark; however, this effect was quantitatively less important than photohemolysis by fenofibrate. On the other hand, fenofibrate photosensitized peroxidation of linoleic acid, monitored by the UV detection of dienic hydroperoxides. Based on the inhibition of this process upon addition of butylated hydroxyanisole, a radical chain (type I) mechanism appears to operate. In summary, fenofibrate is phototoxic in vitro. This behavior can be explained through the involvement of free radicals, singlet oxygen and stable photoproducts. PMID- 8248321 TI - Photolesions and biological inactivation of plasmid DNA on 254 nm irradiation and comparison with 193 nm laser irradiation. AB - Plasmid pTZ18R and calf thymus DNA in aerated neutral aqueous solution were irradiated by continuous 254 nm light. The quantum yields are phi ssb = 4.0 x 10( 5) and phi dsb = 1.4 x 10(-6) for single- and double-strand break formation, respectively, phi br = 2.3 x 10(-5) for base release, phi dn = 2.1 x 10(-3) for destruction of nucleotides, and phi icl approximately phi lds approximately 1 x 10(-6) for interstrand cross-links and locally denatured sites, respectively. The presence of Tris-HCl/ethylenediaminetetraacetic acid (10:1, pH 7.5) buffer strongly reduces phi ssb. The corresponding phi values, obtained on employing pulsed 193 nm laser irradiation, are much larger than those using lambda irr = 254 nm. This is ascribed to a contribution of chemical reactions induced by photoionization, which is absent for 254 nm irradiation. The quantum yields of inactivation of plasmid DNA (lambda irr = 254 nm) were measured by transformation of the Escherichia coli strains AB1157 (wild type), phi ina (1157) = 1.6 x 10( 4), AB1886 (uvr-), phi ina (1886) = 4.2 x 10(-4), AB2463 (rec-), phi ina (2463) = 4.1 x 10(-4) and AB2480 (uvr- rec-), phi ina (2480) = 3.1 x 10(-3). The quantum yields of inactivation of plasmid DNA are compared with those of the four E. coli strains (denoted as chromosomal DNA inactivation) obtained from the literature. The results for E. coli strain AB2480 show that the chromosomal DNA and the plasmid DNA are both inactivated by a single pyrimidine photodimer per genome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248322 TI - Linear dichroism study of metalloporphyrin transition moments in view of radiationless interactions with tryptophan in hemoproteins. AB - We measured the linear dichroism of several metalloporphyrins embedded in stretched polyvinyl alcohol (PVA) films to estimate the orientation of the absorption transition moments, which in hemoproteins are relevant to the radiationless energy transfer between tryptophan and heme. The metalloporphyrins were derivatives of protoporphyrin IX (PPIX), namely Fe(3+)-PPIX (ferric-heme) and Fe2+CO-PPIX (CO-heme), Mg-PPIX (Mg-heme) and Zn-PPIX (Zn-heme). Measurements were conducted between 300 and 700 nm. In all cases the linear dichroism was wavelength dependent, indicating the presence of several transition moments with different orientations. We focused our attention on the near-UV (300-380 nm) and Soret (380-450 nm) absorption bands. Deconvolution in terms of Gaussian components gave three components between 380 and 450 nm and only one in the 300 380 nm region. Deconvolution of the near-UV and Soret spectra of oxy-, deoxy- and carbonmonoxyhemoglobin gave very similar results, suggesting a very similar orientation of the various transition moments in the free and protein-embedded hemes. It should be stressed that the single 300-380 nm band is the only one responsible for the overlap integral that regulates the energy transfer from tryptophan to heme in hemoproteins (Gryczynski et al., Biophys. J. 63, 648-653, 1992). The dichroism of this single band indicated that its transition moment is oriented at about 60 degrees from the alpha-gamma meso-axis of the heme moiety. We conclude that the heme should be considered a linear oscillator when it acts as acceptor of energy transfer from tryptophans. PMID- 8248323 TI - A pulsed laser and pulse radiolysis study of amphiphilic chlorophyll derivatives with PDT activity toward malignant melanoma. AB - Two amphiphilic derivatives of chlorophyll, which have high potential as photodynamic therapy sensitizers for malignant melanoma have been investigated by a combination of laser flash photolysis and pulse radiolysis. It is shown that direct excitation of monomeric forms of these molecules in both hydrophilic and hydrophobic environments produces significant yields of the corresponding triplet states, which have been characterized in terms of spectral and kinetic parameters. In both environments, scavenging of the triplets by oxygen produces singlet oxygen, O2(1 delta g), with essentially unit efficiency as evidenced by time-resolved IR luminescence measurements. PMID- 8248324 TI - Membrane damage induced in cultured human skin fibroblasts by UVA irradiation. AB - Irradiation of cultured human skin fibroblasts with ultraviolet light from 320 to 400 nm (UVA) leads to a decrease in the membrane fluidity exemplified by an enhanced fluorescence anisotropy of the lipophilic fluorescent probe 1-[4 trimethylamino)-phenyl]-6-phenylhexa-1,3,5-triene. This UVA-induced decrease in fluidity is associated with lactate dehydrogenase leakage in the supernatant. Vitamin E, an inhibitor of lipid peroxidation, exerts a protective effect on both phenomena. Therefore, this UVA-induced damage in membrane properties may be related to lipid peroxidation processes. Moreover, exponentially growing cells are more sensitive to these UVA-induced alterations than confluent cells. PMID- 8248325 TI - Photosensitization with etiobenzochlorins and octaethylbenzochlorins. AB - The photophysical and photobiological properties of a series of etiobenzochlorins were evaluated in cell culture using murine leukemia L1210 cells. In the series of agents tested, the chlorin-(mono)sulfonate was the most efficacious, the tin chlorin somewhat less so and the tin chlorin-sulfonate much less active. The parent chlorin was essentially inactive at the limit of solubility. Photodamage was assessed by measuring alterations in surface hydrophobicity (via a two-phase partitioning procedure), amino acid transport and membrane potential. Additional information was provided from fluorescence microscopy, which was used to identify sites of sensitizer binding and effects of photodamage on the binding patterns of fluorescent probes specific for mitochondria, lysosomes and plasma membranes. Effects of photodamage on fluorescence lifetime distribution of the membrane probe trimethylaminodiphenyl hexatriene were examined. The data obtained were consistent with localization of the parnet etiobenzochlorin and tin derivative at lysosomal loci, the chlorin-sulfonate at plasma and mitochondrial membranes and tin-sulfonate at the cell surface. PMID- 8248326 TI - Photoinactivation of influenza virus fusion and infectivity by rose bengal. AB - Rose bengal inactivated influenza virus upon exposure to light. Infectivity and fusion were inactivated with the same dose dependence, supporting the suggestion that the virucidal activity of photodynamic agents against enveloped viruses may be generally due to inactivation of their fusion protein(s). Concentrations required for inactivation were found to depend upon the ratio of rose bengal to virus, rather than on the nominal aqueous concentration. Fusion-competent virosomes were inactivated similarly to intact virus particles. The HA2 portion of the influenza fusion protein HA underwent two different, apparently mutually exclusive modifications upon illumination with rose bengal: cross-linking, and conversion to a form that moved slightly more slowly on sodium dodecyl sulfate polyacrylamide gel electrophoresis. Inactivation of viral fusion was inhibited by oxygen removal or addition of azide or beta-carotene, and was enhanced by D2O, consistent with partial involvement of singlet oxygen. The possibility of a second mechanism of viral photoinactivation, by direct interaction between the viral fusion protein and the photoactivated dye, is also discussed. PMID- 8248327 TI - Measurement of formal potentials for hypericin in dimethylsulfoxide. AB - The formal potentials for hypericin are measured in dimethylsulfoxide using cyclic voltammetry. Two reversible one-electron reductions and a more poorly defined multielectron oxidation are observed. The formal potentials for these processes are consistent with the visible spectrum of hypericin and they indicated that hypericin is both a good oxidizing agent and a good reducing agent in the excited state. PMID- 8248328 TI - Inhibition of 80 kDa protein phosphorylation by short-wavelength UV light in NIH 3T3 cells. AB - The exposure of NIH 3T3 fibroblast cells to 254 nm UV radiation resulted in a temporary depression of DNA synthesis and inhibition of 80 kDa protein phosphorylation. This inhibition of protein phosphorylation was correlated with decreased protein kinase C activity in the membrane fractions of UV-damaged cells. The inositol triphosphate contents measured, by the competitive binding assay using bovine adrenal binding protein, showed 80% reduction in the fibroblasts treated with 15 J/m2 of UV light. The intracellular diacylglycerol concentration was also markedly reduced in UV-damaged cells. The results suggest that UV light causes acute reductions of inositol triphosphate and diacylglycerol contents in cells along with decreases in membrane protein kinase C activity, which leads to the inhibition of phosphorylation of an acidic protein of 80 kDa. PMID- 8248329 TI - Primary DNA damage, HPRT mutation and cell inactivation photoinduced with various sensitizers in V79 cells. AB - DNA strand breaks and hypoxanthine guanine phosphoribosyl transferase (HPRT) mutants were measured in parallel in photochemically treated (PCT) cells and compared at the same level of cell survival. Chinese hamster fibroblasts (V79 cells) were either incubated with the lipophilic dyes tetra(3 hydroxyphenyl)porphyrin (3THPP) and Photofrin II (PII), the anionic dye meso tetra(4-sulfonatophenyl)porphine (TPPS4) or the cationic dye meso-tetra(N-methyl 4-pyridyl)porphine (p-TMPyPH2) before light exposure. In the cells, the lipophilic dyes were localized in membranes, including the nuclear membrane, while the hydrophilic dyes were taken up primarily into spots in the cytoplasm. In addition, the hydrophilic TPPS4 was distributed homogeneously throughout the whole cytoplasm and nucleoplasm. According to the HPRT mutation test, the mutagenicity of light doses survived by 10% of the cells was a factor of six higher in the presence of 3THPP than of PII, whereas for X-rays it was a factor of three higher than for PCT with 3THPP. Light exposure in the presence of the hydrophilic dyes TPPS4 and p-TMPyPH2 was not significantly mutagenic. There was no correlation between the induced rates of HPRT mutants and of DNA strand breaks. Thus, TPPS4 was the most efficient sensitizer with regard to DNA strand breaks when compared at the same level of cell survival, followed by 3THPP, PII and p-TMPyPH2. Hence, the rate of DNA strand breaks cannot be used to predict the mutagenicity of PCT. PMID- 8248330 TI - Protective effect of selenium and zinc on UV-A damage in human skin fibroblasts. AB - Ultraviolet A radiation participates in cytotoxicity and carcinogenesis of the skin by a mechanism involving the generation of reactive oxygen species. Endogenous antiradical defense systems utilize metalloenzymes including Se dependent glutathione peroxidase and Cu and Zn superoxide dismutase. The aim of the present work was to determine the protective effect of two trace elements, Se and Zn, on cultured human diploid fibroblasts exposed to UV-A radiation (broad spectrum source with a maximum intensity at 375 nm). Selenium in the culture medium (0.1 mg/L) in the form of sodium selenite increased the synthesis and activity of glutathione peroxidase by 60.5% in the absence of exposure to UV-A radiation and by 35% after irradiation with 5 J/cm2 (P = 0.043). The presence of this element significantly increased the survival of UV-A-irradiated fibroblasts (P < 0.0001). This confirms the essential role of Se in the detoxifying activity of the enzyme. In addition, thiobarbituric acid-reacting substances (TBAR), which are lipid peroxidation markers, decreased in the presence of exogenous Se: -19% and -22% without irradiation and after irradiation with 5 J/cm2 (P = 0.056). When Zn was added at the dose of 6.5 mg/L as ZnCl2, fibroblasts subjected to oxidizing stress induced by UV-A were protected from cytotoxicity (P < 0.0001). The TBAR production decreased significantly: -33% without irradiation and -34% after irradiation with 5 J/cm2 (P = 0.008). Superoxide dismutase activity, however, decreased after supplementing with Zn: -26% without irradiation and -20% after UV A irradiation (P = 0.017). The antioxidant properties of Zn are thus apparently independent of superoxide dismutase activity. PMID- 8248331 TI - Photocleavage of DNA: irradiation of quinone-containing reagents converts supercoiled to linear DNA. AB - Irradiation (350 nm) of air-saturated solutions of reagents containing an anthraquinone group linked to quaternary alkyl ammonium groups converts supercoiled DNA to circular and to linear DNA. Generation of linear DNA does not occur by accumulation of numerous single-strand cuts but by coincident-site double-strand cleavage of DNA. Irradiation forms the triplet state of the anthraquinone, which reacts either by hydrogen atom abstraction from a sugar of DNA or by electron transfer from a base of the DNA. Subsequent reactions result in chain scission. The quinone is apparently reformed after this sequence and reirradiation leads to double-strand cleavage. PMID- 8248332 TI - Effects of Arctic ozone depletion and snow on UV exposure in Finland. AB - The increase in the UV exposure of the Finnish population associated with the combined effects of ozone depletion and snow reflection was studied with the aid of theoretical calculations based on Green's clear sky UV model. A simple formula was utilized to transform horizontal irradiances to vertical irradiances averaged over 360 degrees azimuth angle. The model was verified with spectral and broadband measurements. The difference between the theoretical and measured UV radiation falling to horizontal surfaces was in most cases less than +/- 10%, and the additional error to theoretical vertical irradiances was less than +/- 10%. The calculations show that the annual horizontal doses in Helsinki (60.2 degrees N, 25 degrees E) are about 35% higher than in Saariselka (68.4 degrees N, 27.5 degrees E), but the difference is only 16% for vertical doses owing to the stronger contribution to vertical (facial) surfaces of the reflection of UV from snow. At Saariselka, the maximum vertical irradiance at the end of April approaches the midsummer values. The ozone depletions up to 40% in February and March 1992 had no significant effect on the annual doses because the total ozone returned to normal before the UV increased to biologically significant levels. PMID- 8248333 TI - An animal model and new photosensitizers for photopheresis. AB - Recently, photopheresis was introduced as a therapy for several T cell-mediated disorders. The treatment results in a specific immune response against the pathogenic clone of T cells involved. However, although promising there is controversy concerning the use of photopheresis in some diseases, e.g. systemic sclerosis. One of the problems is that there is not yet sufficient insight into the mechanism underlying the therapy. This lack of knowledge is partly caused by the fact that there are no easy-to-handle animal models available for photopheresis. This report describes such a model--a Wistar-derived rat with contact hypersensitivity (CHS); a T cell-mediated immune response. White blood cells from CHS rats were simultaneously exposed to 8-methoxypsoralen (8-MOP) and ultraviolet A radiation (UVA) and subsequently intravenously injected into other syngeneic rats suffering from the same disorder. This treatment appears to be very efficacious in suppressing the immunological response against the applied contact allergen, 2,4-dinitrofluorobenzene (DNFB). In addition, the generated suppression of CHS is highly specific and transferable. Furthermore, drugs other than 8-MOP (chlordiazepoxide, nitrofurantoin and chlorpromazine) also appear to be active in our model. PMID- 8248334 TI - In vivo photodynamic effects of phthalocyanines in a skin-fold observation chamber model: role of central metal ion and degree of sulfonation. AB - Six sulfonated metallophthalocyanines, chelated with either aluminum or zinc and sulfonated to different degrees, were studied in vivo for their photodynamic activity in a rat skin-fold chamber model. The chamber, located on the back of female WAG/Rij rats, contained a syngeneic mammary carcinoma implanted into a layer of subcutaneous tissue. Twenty-four hours after intravenous administration of 2.5 mumol/kg of one of the dyes, the chambers received a treatment light dose of 600 J/cm2 with monochromatic light of 675 nm at a power density of 100 mW/cm2. During light delivery and up to a period of 7 days after treatment, vascular effects of tumor and normal tissue were scored. Tumor cell viability was determined by histology and by reimplantation of the chamber contents into the skin of the same animal, either 2 h after treatment or after the 7 day observation period. Vascular effects of both tumor and subcutaneous tissue were strongest with dyes with the lowest degree of sulfonation and decreased with increasing degree of sulfonation. Tumor regrowth did not occur with aluminum phthalocyanine mono- and disulfonate and with zinc phthalocyanine monosulfonate. With the protocol that was used, complete necrosis without recovery was only observed when reimplantation took place at the end of the 7 day follow-up period. Reimplantation 2 h after treatment always resulted in tumor regrowth. At this interval, the presence of viable tumor cells was confirmed histologically. In general tumor tissue vasculature was more susceptible to photodynamic damage than vasculature of the normal tissue. The effect on the circulation of both tumor and normal tissue increased with decreasing degree of sulfonation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248335 TI - Adriamycin resistance in Chinese hamster fibroblasts following oxidative stress induced by photodynamic therapy. AB - Photodynamic therapy (PDT) generates reactive oxygen species that are responsible for the initial cytotoxic events produced by this treatment. An extended (16 h) porphyrin incubation prior to light irradiation increased expression of the 75, 78 and 94 kDa glucose-regulated stress proteins (GRP), as well as the cognate form of the 70 kDa heat shock protein. However, these stress proteins were not induced following isoeffective PDT doses using a short (1 h) porphyrin incubation protocol. In the current study, Chinese hamster fibroblasts were used to examine sensitivity to adjunctive PDT and adriamycin as previous reports indicate a correlation between stress protein synthesis and a decrease in adriamycin cytotoxicity. Treatments that either induced GRP (i.e. PDT with an extended porphyrin incubation or exposure to the calcium ionophore A23187) or did not induce GRP (i.e. PDT with a short porphyrin incubation or UV irradiation) were followed at increasing time intervals with a 1 h adriamycin incubation. A time dependent decrease in adriamycin cytotoxicity was observed when cells were first exposed to either of the PDT protocols or to A23187. Alterations in intracellular drug levels did not account for the change in adriamycin sensitivity. Likewise, intracellular glutathione concentrations and antioxidant enzyme activities were not significantly altered following PDT or A23187. Parameters associated with altered adriamycin sensitivity included a decrease in the percentage of S phase cells following PDT and A23187 as well as a depletion of intracellular ATP after PDT using the extended porphyrin incubation. These results demonstrate that PDT can be added to the growing list of diverse stresses producing transient resistance to adriamycin and that stress protein induction is not universally associated with all oxidative treatments inducing this resistance. PMID- 8248336 TI - Optimization of multifiber light delivery for the photodynamic therapy of localized prostate cancer. AB - The understanding of light distribution within the target organ is essential in ensuring efficacy and safety in photodynamic therapy (PDT). A computer simulator of light distribution in prostatic tissue was employed for optimizing dosimetry for PDT in localized prostatic cancer. The program was based on empirically determined light distributions and optical constants and an assumed fluence rate differential from fiber source to necrosis periphery. The diffusion theory approximation to the Boltzmann transport equation was the applicable formulation relevant to prostatic tissue, which has a high albedo with forward-scattering characteristics. Solving this equation of diffusive transfer for the appropriate fiber geometry yielded the energy fluence distributions for cleaved fiber and cylindrical diffuser light delivery. These distributions, confirmed by our measurements, show a 1/r and 1/square root of r dependency (r = distance from light source) of the fluence phi (r) for the cleaved fiber and diffuser, respectively. This manifests itself by the tighter spacing of energy fluence isodoses in the case of the cleaved fiber. It was predicted that for a typical PDT regime a single interstitially placed cleaved fiber would treat 0.05-0.72 cm3. Four parallel fibers improved the uniformity of light distribution and treatment volume, and an interfiber separation of 12 mm would be necessary to provide optimal overlap of PDT necrosis, treating 0.26-3.6 cm3. The cylindrical diffuser, however, could treat larger volumes, and it was predicted that four 3 cm long diffusers at an optimal separation of 25 mm would treat 25-88 cm3 of prostatic tissue. PMID- 8248337 TI - Sites of photodamage by the iminium salt of a copper octaethylbenzochlorin. AB - Because the benzochlorin derivative copper (II) alpha-meso-N,N' dimethyloctaethylbenzochlorin iminium chloride (CDS1) is not fluorescent, sites of drug localization in L1210 cells were detected by indirect methods involving using a series of fluorescent probes. The CDS1-mediated cytotoxicity was associated with mitochondrial damage, a decreased membrane potential and an increase in the heterogeneity of membrane sites of binding of a polar analog of diphenylhexatriene. Although CDS1 is a cationic compound, its accumulation was not impaired in a cell line exhibiting the multidrug resistance phenotype. PMID- 8248338 TI - Eating patterns of lactating dairy cows of three different breeds fed grass ad lib. AB - Eating behavior of lactating cows of three breeds (Holstein-Friesian = HF, Simmental = SI, and Jersey = JER) was compared in the present study. The cows were kept in a loose housing system and fed grass and hay ad lib along with concentrates. Within 11 days, 1396, 1504, and 1682 grass meals were recorded and analyzed for HF, SI, and JER cows, respectively. In cows of all three breeds, 89% of the grass meals (= 94% of the total grass intake) occurred during the light phase (0400-2200) and peaks of grass intake with large meals were always observed shortly after refilling the troughs. A similar diurnal distribution was evident for concentrate intake, but was less pronounced (28% eaten during the dark phase), and peaks of concentrate intake occurred also in the evening and after midnight. Hay intake was negligible and was not considered for meal pattern analysis. JER cows ate generally less than HF or SI cows, but spent more time eating. Grass meal frequency was higher, but grass meal size and mean eating rate within grass meals was smaller for JER cows than for HF and SI cows. Significant correlations between the size of grass meals and the duration of either the interval preceding or following that same meal were observed only for some animals (in particular JER and SI cows) during the main eating period in the morning (0800-1300), but barely in the afternoon (1300-1700). Thus, the short term physiological control of meal-taking in dairy cows kept under normal farming conditions seems to be markedly influenced by external factors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248339 TI - Independence of genetic influences on body size, daily intake, and meal patterns of humans. AB - Body size and daily and meal nutrient intakes have been shown to be affected by heredity. The present study investigated the independence of the heritability of overall daily intake from body size, meal intake from overall daily intakes, and meal macronutrient intake from meal size. Previously collected 7-day food intake diary data obtained from 109 identical and 86 fraternal adult twin pairs, who lived separately, were reanalyzed. A series of linear structural models were fit, using maximum likelihood estimation, to the identical and fraternal twin covariance matrices for the body size, overall intake, and meal intake. Gender and age were used as additional covariates. The results suggest that 96% of the variance in height and 85% of the variance in weight are due to heredity. They also indicate that from 25% to 50% of the variance in daily food intake and from 22% to 48% of the variance in average meal size is due to genetic influences independent of the body size, gender, and age of the individual. Furthermore, they indicate that from 18% to 42% of the variance in average meal size is due to genetic influences independent of the overall daily intake, gender, and age of the individual. On the other hand, the influence of heredity on the macronutrient composition of the meals was found to result, for the most part, secondarily from the genetic influence on overall meal size. These results strongly suggest that there are separate genetic influences on body size, overall intake, and meal intake that have interacting influences on one another.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248340 TI - The caloric demand of lactation does not alter spontaneous meal patterns, nutrient intakes, or moods of women. AB - The regulation of food and fluid intake in lactating women was examined by comparing maternal intake during lactation, 6 weeks postpartum, to intake in two groups of nonpregnant, nonlactating women. All subjects completed a questionnaire and then filled out a food-intake diary for seven consecutive days. Lactating women did not differ from body weight-matched, nonlactating controls in their total daily intakes or their meal patterns. However, they consumed a significantly smaller percentage of the recommended dietary allowances (RDA) per day than did their nonlactating counterparts. Lactating women were also significantly more calm both pre- and postmeal than were either of the control groups. These results indicate that the lactating women, at 6 weeks postpartum, did not increase their intake to compensate for the caloric demands of lactation. This may indicate that the lactating women catabolize weight gained during pregnancy faster than accounted for in the RDA, or that lactating women increase their metabolic efficiency. PMID- 8248341 TI - Feeding behavior and an oropharyngeal component of satiety in a two-headed snake. AB - We studied the feeding behavior of a dicephalous Black Rat Snake (Elaphe o. obsoleta) whose heads attack and ingest prey independently. Both heads were given satiation feedings with mouse prey at either 5-day intervals (1989, n = 16 feedings) or 9-11 day intervals (1990, n = 4 feedings). In 1989, a meal pattern was manifested as a negative correlation between total (left + right) meal size (g) and total size of the last meal. Meal sizes of the heads were consistently different (L > R) despite their apparently identical genotypes. We confirmed tongue-flicking rate and swallowing time as indices of satiety/hunger in each head. In order to test whether oropharyngeal stimulation is involved in satiation, we analyzed three measures: willingness to feed, and intrameal changes in swallowing, and tongue-flicking rates. Data for these tests were taken from experimental feedings in which one head was fed to satiation (first head) before the other head (second head) was fed. The second head generally did eat (8 of 10 experimental feedings) after the first head had become satiated. In addition, while the time to swallow a mouse generally increased as the snake neared satiety, swallowing time decreased significantly (p = 0.04) from the first head's last mouse to the second head's first mouse. Finally, in two of three feedings, the first head's tongue-flicking rate decreased before the second head's. These results, which take advantage of this unique animal, indicate that oropharyngeal stimulation is involved in meal termination. PMID- 8248342 TI - Effects of vagotomy on Leu-enkephalin-induced changes in memory storage processes. AB - When given before or after training, Leu-enkephalin impairs later retention of learning. To replicate these findings, Experiment 1 determined if Leu-enkephalin impairs retention when administered after inhibitory avoidance training. Posttraining injection of 100.0 micrograms/kg of Leu-enkephalin impaired retention relative to saline controls or those receiving a lower dose of this peptide. Since Leu-enkephalin does not cross freely from the blood stream into the brain, this peptide may exert its influence on mnemonic processes by activating peripheral receptors that transmit neural messages to the brain via the vagus nerve. In Experiment 2, 100.0 micrograms/kg Leu-enkephalin impaired retention in unoperated and sham-operated animals although vagotomized animals did not differ significantly from these groups or saline controls. These results suggest that subdiaphragmatic vagotomy may not be sufficient to disrupt the mnemonic effects of Leu-enkephalin. Alternatively, the vagus nerve may be one pathway but not the only pathway by which the activation of peripheral systems may influence the memory storage process. PMID- 8248343 TI - Circadian rhythms of food and water intake and urine excretion in diabetic rats. AB - The light/dark (L/D) rhythms of food and water intake and urine output were studied in normal and diabetic rats for 7 consecutive days at 4-h intervals. The control rats showed the highest values of these parameters during the dark phase (83.68% food, 68.71% water, and 67.44% urine). The diabetic rats also maintained this nocturnal predominance, although the values were less marked (59.55% food, 55% water, and 56% urine). A circadian rhythm of food (phi = 3.31 h) and water (phi = 3.54 h) intake as well as of the volume of urine excreted (phi = 1.10) was detected in the control animals. The diabetic rats, inspite of presenting polyphagia, maintain the circadian rhythm of food intake, whereas a loss of the normal circadian variation of drinking intake was observed as well as the absence of circadian rhythm in urinary excretion. It was concluded that in streptozotocin diabetic rats we have observed a loss of the normal patterns of water intake and urinary excretion, perhaps masked by the polydipsia and polyuria, whereas the circadian rhythm of food intake remains. PMID- 8248344 TI - The effects of fludrocortisone acetate and deoxycorticosterone acetate on salt appetite in mice. AB - Three genotypes of mice were shown to increase their intake of 3% sodium chloride (NaCl) following administration of fludrocortisone acetate (25 mg/kg), a synthetic steroid having strong mineralocorticoid/glucocorticoid activity. However, treatment with deoxycorticosterone acetate (DOCA) (10 mg/kg), a corticosteroid with predominantly mineralocorticoid activity, did not increase 3% NaCl consumption by the stock tested. In contrast, both fludrocortisone and DOCA have been shown to be effective activators of sodium appetite in rats. The essential importance of glucocorticoids in potentiating mineralocorticoid activation of sodium hunger in mice is suggested. PMID- 8248345 TI - A twin study of genetic and environmental influences on the intake of fluids and beverages. AB - The heritability of fluid and beverage intake was investigated with 109 identical and 86 fraternal adult twin pairs who were paid to maintain 7-day diaries of everything they either ate or drank. Both classical analysis of heritability and linear structural modeling revealed that the amount of fluids ingested daily or in bouts is strongly influenced by heredity in males and by familial environment in females. This was true for the total amount of water ingested, regardless of source, the amount of fluid ingested in excess of digestive requirements, and the amount ingested in the form of drinks. Linear structural modeling revealed that these influences act primarily on the overall total intake of fluids and affect drink fluid intake only indirectly. Significant influences of the genes and the environment were also found on the ingestion of particular beverages, independent of the effects of age and the total intake of fluids. In particular, significant genetic influences were identified on the intake of alcohol, sugared soda, coffee, and milk, and significant familial environmental influences on the intake of diet soda and fruit juice. These results suggest that both the amounts and the types of fluids ingested are complexly influenced by heredity and the environment, depending upon the gender of the individual and the type of beverage investigated. PMID- 8248346 TI - Sex differences in open-field behavior in response to the beta-carboline FG 7142 in rats. AB - Sex differences in animal models of anxiety and depression that employ external stimuli have been previously reported. This study examined the effect of gender on pharmacologically induced anxiety in rats coupled with novel handling, injection, and activity in an open-field test. The anxiogenic compound FG 7142 significantly decreased male open-field exploratory at 5, 10, and 20 mg/kg, while rearing behavior was decreased only at the 20 mg/kg dose. Female rats were more resistant to the effects of FG 7142 on open-field exploratory behavior, decreasing open-field activity, and rearing behavior only after the administration of a dose of 40 mg/kg. In addition, a significant sex difference was observed in the open-field activity of rats injected with the vehicle control. Male rats were less active but showed a similar level of rearing behavior when compared to female rats. The greater sensitivity of male rats to the activity-suppressant effects of FG 7142 could explain the sex differences observed in several other animal models of anxiety and depression. PMID- 8248347 TI - Augmenting and reducing of visual evoked potentials in Roman high- and low avoidance rats. AB - Human and cat high sensation seekers tend to show increasing amplitudes (augmenting) of the P1 and N1 components of the visual evoked potential (VEP) to increasing intensities of light flash, whereas low sensation seekers show VEP reducing. Roman high-avoidance (RHA) and Roman low-avoidance (RLA) rats have behavioral traits comparable to human and cat high and low sensation seekers, respectively. RHA rats show greater exploration, activity, and aggression than do RLA rats. Rats of each Roman line and Wistar rats were anesthetized with chloral hydrate and maintained at a stable moderate anesthetic level. VEPs to each of five flash intensities were computer averaged for each rat. The slopes of P1 amplitudes as a function of flash intensity were significantly greater in the RHA than the RLA rats. RHA rats were clear augmenters; RLA rats had almost flat amplitude-intensity functions. The Wistar rats had slope functions that were similar to those of the RLA rats. This study shows that the relationship between sensation-seeking behavior and VEP augmenting and reducing has a heritable base and extends across species from human, cat, and rat. In addition, we demonstrate a rat model of this relationship that yields advantages of genetic homogeneity and a short generational time, and provides access to a wealth of behavioral data and experimental manipulations available for the rat. PMID- 8248348 TI - Ontogeny of immobility and response to alarm substance in the forced swim test. AB - The ontogeny of immobility in the forced swim test and the response to alarm substance were determined in rats. Immobility, defined as floating vertically in water, making only those minimal movements necessary to keep the head above water, emerged at 21 days of age and stabilized beginning at 26 days (< or = 10% variation from the mean time of the previous day). Males and females did not differ in maturation of this response. The reaction to alarm substance was discernible when animals were 24 days of age. Again, males and females were not significantly different in ontogeny of this response. PMID- 8248349 TI - The effects of the acute administration of RU 486 on dietary fat preference in fasted lean and obese men. AB - The effects of RU 486, a potent glucocorticoid antagonist, on dietary fat preference were explored in obese men and lean controls in a double-blind crossover study. An oral 10 mg/kg dose of RU 486 or placebo was administered at midnight the second night of a 48-h hospital stay. Macronutrient and caloric intakes were calculated each day and a taste test of six commercial dairy products (fat content by weight < 0.5%, 2.0%, 3.3%, 10.5%, 18%, and 36%) was performed. Dairy products were judged for pleasantness, creaminess, and overall preference. Subjects were then asked to consume their favorite dairy product until sated. Urinary free cortisol (UFC) and plasma adrenocorticotropic hormone (ACTH), cortisol, insulin, and glucose were determined. Intake of a self-selected diet was recorded. As expected, in response to RU 486, UFC increased from 120 +/- 25 micrograms/24 h to 297 +/- 73 micrograms/24 h (p < 0.05) in obese men and from 81 +/- 10 micrograms/24 h to 357 +/- 109 micrograms/24 h (p < 0.05) in lean men. Plasma cortisol increased from 26.1 +/- 1.1 microgram/dl to 31.8 +/- 1.0 microgram/dl (p < 0.05) in obese men and from 26.1 +/- 1.7 micrograms/dl to 32.2 +/- 1.7 micrograms/dl (p < 0.05) in lean men. Plasma insulin was significantly higher in obese 24.6 +/- 3.2 microIU/ml than in lean men 12.8 +/- 1.1 microIU/ml (p = 0.0001) but was unaffected by RU 486. RU 486 did not decrease fat intake in either obese or lean men.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248350 TI - Effects of repeated stress on plasma arginine levels in young and old rats. AB - The effect of age and recurring stress responses on endogenous plasma arginine (ARG) concentrations were determined. Blood from catheterized young (3 months) and old (24 months) were used to ascertain the plasma concentrations of ARG under baseline conditions, during a 30-min immobilization experience and during a 3.5 h poststress period. Rats were again immobilized and blood drawn following a 1 day rest period (day 3) and then after an additional 3 day rest period (day 7). For young animals, ARG levels decrease significantly after 30 min of stress and remain suppressed during the entire poststress period (3.5 h). After a 1 day rest period, these young rats may have exhibited adaptation to stress on day 3. Old animals showed little or no reduction in plasma ARG concentrations during and after immobilization stress exposure on all three occasions. Baseline levels of old animals were lower in comparison to young animals and their stress responses were smaller. Thus, it is possible that significant changes in plasma ARG levels (as the nitric oxide precursor) during or after stress could serve as an indicator of impending pathology such as cardiovascular disease or immunocompetency. PMID- 8248351 TI - Influence of social isolation, gender, strain, and prior novelty on plus-maze behaviour in mice. AB - Behavioural baseline is a critical determinant of response to drugs and other manipulations. In the present study, the influence of several organismic and procedural variables on basal plus-maze profiles in mice were examined. The methodology incorporated traditional behavioural parameters as well as novel measures derived from ethological analysis. Experiment 1 showed that social isolation for 1-3 weeks enhanced aggression in male DBA/2 mice but did not substantially alter their behaviour on the maze. A reduction in stretch attend postures did, however, suggest a minor reduction in anxiety in socially isolated animals. In Experiment 2, males of both DBA/2 and T1 strains exhibited higher levels of general activity on the maze than their female counterparts. Although additional evidence suggested that DBA/2 (but not T1) females were less anxious than males, no major sex differences were noted. Experiment 3 revealed a significant strain difference in plus-maze profiles, with T1 males showing a lower basal level of anxiety than DBA/2 males. This study also demonstrated that DBA/2 and T1 males react very differently to prior novelty experience, with enhanced anxiety evident in the former and reduced anxiety in the latter. Together, these findings point to a range of organismic and procedural variables that may account for inconsistencies in the literature on the elevated plus-maze. PMID- 8248352 TI - An iodinated vasopressin (V1) antagonist blocks flank marking and selectively labels neural binding sites in golden hamsters. AB - An arginine-vasopressin (AVP) derivative, [d(CH2)5,Sar7]AVP (SAVP), has been characterized as an antagonist to vasopressin V1 receptors. Using AVP-dependent flank-marking behavior as a bioassay, it was possible to verify that iodinated SAVP (I-SAVP) retains biological activity within the central nervous system, as the antagonist blocked the behavior. Furthermore, 125I-SAVP was used to localize specific V1 binding sites in the brain. The resulting binding was localized to discrete anatomical sites, and highly specific to V1-like receptors. While we confirmed previous findings using 3H-AVP in golden hamsters, we also identified binding in many areas previously unreported (e.g., arcuate and paraventricular nuclei of the hypothalamus, tenia tecta, posteromedial cortical nucleus of the amygdala, and zona incerta), suggesting that 125I-SAVP provides a greater level of resolution. In addition, specific binding was observed in the lateral septum, anterior hypothalamus, and midbrain central gray, areas that have previously been shown to trigger flank marking in response to AVP microinjection. The presence of AVP binding sites in limbic and mesencephalic areas involved in the regulation of flank marking suggests that this neuropeptide may play an important role as a neurotransmitter at multiple levels in the neural circuits controlling this behavior. PMID- 8248353 TI - Effects of early protein malnutrition and repeated testing upon locomotor and exploratory behaviors in the elevated plus-maze. AB - An elevated plus-maze was used to investigate the effects of repeated testing on the locomotor and exploratory behaviors of malnourished rats. Pup malnutrition was induced during the lactation period (0 to 21 days of age) by feeding the dams a protein-deficient diet (6% protein) and the animals were allowed to recover from weaning to 70 days of age by eating a commercial lab chow diet. Control animals were suckled by dams receiving a normal protein diet (16% protein) during the lactation period and were fed a commercial lab chow diet after weaning. At 70 days, malnourished and control animals were placed on the central platform of the elevated plus-maze facing an enclosed arm and allowed to explore for 5 min. This procedure was repeated at 24-h intervals for 6 days. The repeated testing in the elevated plus-maze did not change the total number of arm entries and attempts to enter open arms, but decreased the percentage of open arm entries, time spent in open arms, and total time spent on the central platform. These data suggest an increase in anxiety with repeated testing in the elevated plus-maze. In addition, the malnourished animals showed a larger number of both rearings and attempts to enter the open arms, suggesting a high level of exploration and/or high impulsiveness of these animals as compared to control. The elevated plus-maze proved to be a useful animal model to evaluate exploratory behaviors in early protein malnourished animals. PMID- 8248354 TI - Hyperphagia and obesity in female rats with temporal lobe lesions. AB - Damage to the temporal lobes in cats, dogs, and primates has long been known to result in hyperphagia and obesity, but research into the role of this area of the brain in feeding behavior has largely been neglected because of an inability to produce similar results in rats. The present study reports hyperphagia and obesity in female rats with small electrolytic lesions centered in the posterodorsal amygdala. Daily food intake more than doubled in the first few days after surgery and mean weight gain was more than four times that observed in animals with sham lesions during the first 26 days. The rats with lesions were not hyperresponsive to a switch in diets (lab chow to high-fat, and back). In all animals that gained abnormal amounts of weight, the posterior extent of the lesions extended through the amygdalohippocampal area into the ventral hippocampal formation. The results suggest that the temporal lobe is an important extrahypothalamic site for the regulation of food intake in rodents. PMID- 8248355 TI - Psychoneuroendocrine outcomes of short-term crowding stress. AB - One-week exposure to crowding stress has been reported to affect body weight and pituitary-adrenal axis activity. This study has first evaluated whether the aforementioned changes are associated with altered exploratory activity. One-week crowding (16 rats/cage) diminished body weight gains, compared to the control situation (four rats/cage): actually, this difference was mainly due to a marked loss in food intake and body weight following the first day of crowding. On the other hand, 1-week crowded rats were not different from their controls when placed for the first time in an open field, as shown by the analysis of their exploratory activities, their number of grooming episodes, and their defecation. Psychoneuroendocrine effects of 1-day crowding were then investigated: actually, open field behaviors were not modified but basal plasma glucose levels were decreased. Moreover, neither plasma ACTH and corticosterone levels nor 2-min cold swim-induced increases in corticosterone levels were affected. Besides, cold swim induced increases in plasma ACTH levels were amplified in crowded rats, and these increases were associated with hyperglycemia in crowded, but not in control rats. It is suggested that 1-day exposure to crowding has differential consequences upon feeding and exploratory behaviors while potentiating the responsiveness of stress hormones. PMID- 8248356 TI - Effects of voluntary exercise on immune function in rats. AB - We investigated effects of voluntary wheel running on specific antibody responses to Keyhole Limpet Hemocyanin (KLH) and on mitogen-stimulated lymphocyte proliferation in adult male rats. Each subject was placed in a running wheel for 12 h daily during the dark portion of the light cycle for a total of 8 weeks. For experimental animals the wheels rotated freely, enabling subjects to exercise at will, while for control animals the wheels were prevented from rotating. Subjects were immunized with KLH (25 micrograms) 5 weeks into the study, and blood samples were collected intermittently from the tail for 3 weeks and later assayed for anti-KLH antibody levels. At the end of the study, subjects were sacrificed and spleens were dissected and assayed for lymphocyte proliferative responses to the mitogen Concanavalin A. Exercised rats gained less weight and had higher splenic proliferative responses than control rats; however, there were no significant group differences in anti-KLH antibody levels. PMID- 8248357 TI - Aberrant transients in the EEG of epileptic rats: a spectral analytical approach. AB - Aberrant transients in the cortical electroencephalogram of rats of the epileptic WAG/Rij strain were studied by means of spectral analysis. The EEG of rats of this strain contains, besides normal sleep spindles, high-voltage spiky phenomena, epileptic spike-wave discharges, and deviant intermediate stage. Spectral analysis of these transient phenomena shows that some features, like their peak frequency, are alike, but that they differ in other spectral characteristics, as in the first harmonic of the peak frequency and in the domain of the high frequencies. The results provide arguments for the view that spike wave discharges might be considered as unique aberrant phenomena, presumably related but dissimilar to the high-voltage spiky phenomena and intermediate stage. Next to this, spectral analysis was used to study the intraphenomenal dynamics of spike-wave discharges. The peak frequency was found to decrease monotonously from about 10 Hz at the beginning of the spike-wave discharge to about 8 Hz at the end. Other spike-wave discharge frequency bands showed an intraphenomenal increase followed by a decrease. These time-variant EEG dynamics in spike-wave discharges might correlate with the cognitive disturbances during absence seizures in man. PMID- 8248358 TI - Evaluation of the stability and quality of sleep using Hjorth's descriptors. AB - Hjorth's descriptors (NSD) (activity, mobility, and complexity) provide a useful tool for evaluating micro- and macrostructural elements of sleep. In rats, the automatic analysis of sleep recordings by means of NSD calculated from SM and Vis derivations allow different sleep stages to be discriminated. Activity distribution over the total recording permits the definition of a global effect on the EEG. Moreover, the quality of sleep can be evaluated by the variations of the SM activity distribution by considering different classes of amplitude. Index and rate of unstable amplitude segments (UAS) constitute useful parameters to analyse the stability and quality of sleep with different models of insomnia and after pharmacological treatment. The UAS in rats can be compared to CAPs in humans. The NSD are also able to define quality and stability of human sleep. EEG analysis using NSD provides a novel perspective for the analysis of the stability and quality of rat and human sleep. This microstructural analysis of sleep appears to be a useful tool for pharmaco-EEG studies. PMID- 8248359 TI - Arousals without awakening--dynamic aspect of sleep. AB - K complex as an elementary form of arousal was investigated by evoked K complexes. The components building up evoked K complexes and topographical sleep level differences and influence of the kind of cognitive elaboration were analysed. Evidences are presented supporting that K complex represents a cascade of events originating from sources of different topography and probably by a different kind of elaboration activated in a certain order requested by the nature and context of the eliciting stimuli. The power spectra of evoked arousals -including K complexes--were investigated. The poststimulus spectral pattern is characterized by a short initial power elevation and a following reduction of all frequency bands except a simultaneous but prolonged (5-20 s) and strong (50%) power reduction at the 13-14 Hz sigma spindle band. This phenomenon seems to be a common feature in different stages of slow wave sleep. This stimulus-related microstate could serve as a transitory stand-by state ready to reach higher arousal rapidly while maintaining the continuity of sleep; hence, the inhibition of spindle activity could provide a phasically improved thalamocortical sensory inflow after environmental stimuli. A microstructural scheme of arousals without awakening has been delineated. PMID- 8248360 TI - Aging and changes in phasic events during sleep. AB - Changes in phasic events in the elderly are reviewed. Such phasic events may in part be determined by the macrostructure of sleep (sleep stages). Therefore, a brief description of sleep architecture and EEG morphology in the aged person is given. Second, there is a marked variability among individuals in the number of spindles and K-complexes and more so in older individuals than in younger. However, there is an overall decrease in these events with age. The characteristics of these events (amplitude, frequency content, and distribution) change simultaneously. Third, the number of rapid eye movement occurring during REM sleep decreases with aging, but important gender differences exist, in that women generally maintain a higher REM density. The degree of changes in phasic events might correlate with mental deterioration (sleep cognition theory), but physical factors might also be the underlying reason for the observed changes. Whether these changes are indicative of the individual's ability to maintain sleep remains to be determined. PMID- 8248361 TI - Clinical applications of cyclic alternating pattern. AB - Sleep-related disorders are revisited in the light of the physiological modality of NREM sleep expressed by the cyclic alternating pattern (CAP). Owing to its fluctuating properties on vigilance, muscle tone, and vegetative activities, CAP represents a highly favorable condition for the occurrence of interictal generalized and focal lesional EEG discharges, for motor seizures, and for periodic jerks in nocturnal myoclonus. All these manifestations are significantly associated with the components of activation during CAP, i.e., the A phases. On the contrary, the B phases of CAP appear chronologically linked to inhibitory phenomena in epileptic patients and in nocturnal myoclonus. The two phases of CAP seem especially involved in sleep apnea syndrome, where respiration is interrupted during a phase B and restored during a phase A. CAP rate, that measures the effort of the brain to maintain sleep, is increased by all conditions that induce vigilance instability such as noise, clinical insomnia, interictal EEG paroxysms, nocturnal seizures, periodic leg movements, and in certain extreme pathologic conditions such as Creutzfeldt-Jakob disease and stage 2 coma. PMID- 8248362 TI - Shifts and awakenings: valuable indicators of the quality of sleep: do these parameters react differently to hypnotics? AB - In order to test the assumption that the sleep stability increases as the number of shifts and awakenings decreases, all-night sleep recordings (registered at home) were reanalyzed. The registrations belong to 10 young good sleepers, 10 adult good sleepers, 10 insomnia patients without treatment, 10 treated with a benzodiazepine and 10 treated with a nonbenzodiazepine hypnotic, 10 patients with proven narcolepsy, and 10 with obstructive sleep apnea syndrome (OSAS). For each of the categories NREM-1-2, NREM 3-4, and REM sleep, indices were defined as the number of shifts and awakenings that occurred per hour. Our results show that the indices of awakenings for the good as well for the bad sleepers were similar, whereas the indices of awakenings were very group dependent. We interpreted these results as showing that awakenings are valuable indicators for studying the quality of sleep, whereas shifts are not. PMID- 8248363 TI - Sleep phasic events in dysthymic patients: a comparative study with normal controls. AB - Among psychiatric entities, dysthymic patients provide a good model for studies on minor disturbances in sleep organization because they present borderline abnormalities in their night sleep that concern mainly slow wave sleep. A study on sleep phasic events was performed to evaluate abnormalities in sleep microstructure. Therefore, for all subjects--normal controls and dysthymic patients--the following phasic activities were scored: K-complexes, vertex sharp transients, posterior occipital transients, delta paroxysms, theta bursts, microarousals, and awakenings. Furthermore, other sleep EEG patterns, such as REM with spindles, and intermediate stages, were assessed. The following results were obtained: 1) dysthymic patients have less K-complexes and vertex sharp transients, with lower densities of these events; 2) they have a reduced number of microarousals, with an increased rate of awakenings; 3) patients present a higher incidence of other events such as theta bursts, POSTs, REM with spindles, and intermediate stages. These results suggest that the analysis of phasic events is useful for the discrimination between normal and pathologic sleep, even when there are only minor differences in terms of macrostructure. PMID- 8248364 TI - Stimulation of drinking by bacterial endotoxins in the rat. AB - We investigated the effects of endotoxins on water balance, rectal temperature, and food intake in male Long-Evans rats with femoral venous catheters. Extracts of Escherichia coli or Salmonella minnesota, in doses ranging from 125 to 500 micrograms/kg IV, stimulated drinking and reduced urinary water loss for several hours. The net gain of 5 ml water 2 h after the lowest dose of E. coli endotoxin was sufficient to reduce plasma osmolality and sodium concentration 2 to 3%. Drinking occurred during the period of hypothermia that frequently precedes the onset of endotoxin-induced fever, so cannot be attributed to increased body temperature. Doses of endotoxin causing drinking inhibited both spontaneous and deprivation-induced feeding. The cause of the drinking is not known, but may involve mechanisms other than the known dehydrational signals controlling thirst. PMID- 8248365 TI - Relationship between emotional status and spontaneous behavior in rats exposed to high pressure. AB - When humans and experimental animals are exposed to increased environmental pressure, they develop the high pressure neurological syndrome. In the present study, we have investigated the relationship between the emotional status and the development of pressure-induced behavioral disorders, such as locomotor and motor activity (LMA) and myoclonia, in two genetically selected lines of rats (Roman low-(RLA/verh) and high-(RLA/verh) avoidance), which differ drastically in their level of emotionality. RLA/verh rats presented a greater level of LMA than RHA/Verh rats; RLA/Verh rats also showed myoclonia while RHA/Verh rats did not. These results are discussed in the light of recent experiments in rats exposed to stressful situations or pressure environment. It is concluded that the use of both RLA/Verh and RHA/Verh rats would be of particular interest to better understand the neurochemical and neurological processes underlying the development of LMA and myoclonia in rats exposed to high pressure. PMID- 8248366 TI - Nutritional status, suckling behavior, and prolactin release during lactation. AB - This experiment was conducted to determine whether suckling-induced prolactin (PRL) values in chronically food-restricted lactating rats resulted from the inability of their litters to suckle vigorously. Rats were randomly assigned to be fed ad lib (AL) or to be chronically restricted (CR) to 50% of AL before and during pregnancy and lactation. Rats nursed their own litter (own) or a litter of the opposite dietary treatment group (other) for 90-min periods at early or peak lactation. Blood was collected regularly through a catheter implanted several days earlier. Suckling contributed to differences in PRL between AL and CR dams in early lactation as demonstrated by an interaction between dietary treatment and type of litter. However, tests of AL own vs. AL other and CR own vs. CR other were not significant. This is probably because the inattentive behavior of CR dams attenuated the PRL response in this group. At peak lactation, PRL values were related to recovery time from catheter implantation; therefore, determinants of PRL release change over the course of lactation. PMID- 8248367 TI - Metabolic correlates of behaviour in the newborn infant. AB - Twenty-four infants were submitted to behavioural observation 0.5 and 3.5 h after birth. Blood samples were collected from the umbilical cord and from the infant immediately following observation, for the determination of glucose, insulin, cholecystokinin (CCK), and free fatty acids (FFA). It was found that 88% of the infants were awake and 83% performed sucking movements 0.5 h after birth; these findings were associated to a decrease in the plasma glucose concentration (p = 0.05). Eighty-eight percent of the infants were asleep and only 12% exhibited sucking movements 3.5 h after birth; at this time point there was an increase in the plasma concentrations of both CCK (p = 0.05) and FFA (p = 0.01). The infants' plasma insulin concentrations remained unchanged during the time course. It is suggested that the initial behavioural pattern of the newborn, characterized by the appearance of sucking movements, may be due to the rapid decrease of the circulating glucose concentration. The successive increase in the blood concentrations of both CCK and FFA might be two of the factors contributing to the cessation of sucking and to the establishment of a sleeping state. PMID- 8248368 TI - Brain-reactive antibodies and behavior of autoimmune MRL-lpr mice. AB - The hypothesis that brain-reactive autoantibodies (BRA) impair behavior was examined in MRL-lpr mice, which develop spontaneous autoimmune disease. Circulating BRA were measured as in vitro serum reactivity to Neuro-2A neuroblastoma cell line, and behavioral competence was assessed in activity monitors, open field, beam walking, and Morris water maze task. Mice with BRA in serum (BRA positive) exhibited slower spontaneous locomotion in a novel environment, shorter grooming episodes, and less exploration of the open field centre when compared to age-matched 7-11-week-old BRA-negative cagemates. Moreover, when initially exposed to the large swimming pool, BRA-positive mice showed increased swimming along the wall, but had no difficulty in learning the water maze task or in traversing a narrow beam. Brain-reactive autoantibodies titre and behavioral measures were not correlated, suggesting that the concentration of serum BRA is not reflective of the magnitude of behavioral impairment. Nevertheless, the present study suggests that the presence of circulating BRA is associated with impaired exploration and/or enhanced emotional reactivity in MRL-lpr mice. It also supports the hypothesis of a pathogenic role of BRA in various mental disorders. PMID- 8248369 TI - Chronic intrahypothalamic insulin infusion in the rat: behavioral specificity. AB - This study examines whether chronic intrahypothalamic (IH) insulin infusions suppress body weight and food intake directly or via effects on water intake or activity. Insulin (15 microU/h) was infused into the ventromedial hypothalamic nucleus of rats for 1 week. If IH insulin infusions primarily suppress water intake, animals should consume less water during insulin infusion in the absence of food. In the first experiment in this study, rats food deprived during IH insulin infusion did not drink significantly less than during vehicle infusion. This implies that IH insulin affects water intake secondarily to its impact on food intake. Insulin might suppress food intake and body weight by decreasing overall activity levels, including activity involved in ingestive behavior. In the second experiment, rats' activity on a running wheel was measured during IH insulin and vehicle infusion; activity increased during insulin infusion compared to vehicle infusion. These findings suggest that insulin's effects on food intake and body weight are via a mechanism that does not appear to directly influence water intake, and does not reduce overall activity levels. PMID- 8248370 TI - New variables for defining sleep continuity. AB - One very synthetic way to represent a night's sleep is by way of a hypnogram: a graphical representation of the sleep stages as a function of time. The hypnogram is generally quantified by a series of variables that measure the durations and latencies of the various sleep stages including wake. These variables, however, do not fully account for all the information contained in the hypnogram, in particular information on sleep continuity. A series of variables that measure and localize disruption of this continuity are proposed and their utility validated on three groups of patients presenting sleep disorders. Utility is established if the variable is capable of differentiating between patients and healthy controls. Two sets of variables are examined: those that use the entire sleep period as unit of measurement, and those that are measured within each consecutive NREM-REM sleep cycle. The results show that the variables proposed are able to differentiate between groups and, therefore, are useful measures reflecting the hypnogram more precisely. They also show that fragmentation of REM sleep does not present a systematic trend across the night, but that fragmentation of NREM sleep goes up linearly. PMID- 8248371 TI - Short-term effects of different amounts of sweet and nonsweet carbohydrates on satiety and energy intake. AB - If a sweet taste sensation increases appetite it can be hypothesized that sweet carbohydrates (CHOs) are less satiating than nonsweet CHOs. We studied this hypothesis in two experiments. In two within-subjects repeated measures experiments, subjects (n = 28 in study 1, n = 29 in study 2) got lunches of varying energy content. There were five energy levels (250, 350, 450, 550, and 650 kcal) in the first experiment, and three energy levels (250, 500, and 750 kcal) in the second experiment. Within each energy level there was one lunch with sweet, and one lunch with nonsweet CHOs. The weight, protein, fat, carbohydrate, and fibre level was kept constant within each energy level. The results of the first study confirmed the hypothesis that sweet CHOs were less satiating than nonsweet CHOs. The results of the second study did not show a difference between the satiating effect of sweet and nonsweet CHOs. There was a strong sensory specific effect in both studies. Sweet CHOs had a suppressing effect on appetite for something sweet, whereas nonsweet CHOs suppressed appetite for something savoury. In the second study, sweet CHOs suppressed the intake of sweet food items in a test meal 2 h later in a dose-response manner. Nonsweet CHOs suppressed the intake of nonsweet food items, also in a dose-response manner. It is concluded that energy (or a food property related to energy) has a strong effect on appetite. The taste had a sensory specific effect on subsequent appetite and food selection. PMID- 8248372 TI - Passive-avoidance behavior in Wistar-Kyoto (WKY), Wistar, and Fischer-344 rats. AB - In Experiment 1, Wistar-Kyoto (WKY), Wistar, and Fischer-344 (F-344) rats were tested on two passive-avoidance tasks: a platform step-down task and a one-way avoidance task. Wistars had shorter response latency scores in the step-down task, whereas F-344 rats had shorter scores in the one-way task. A subsequent ulcerogenic water-restraint stress produced significantly fewer ulcers in Wistar rats as compared to WKY and F-344 rats. In Experiment 2, rearings, grooming, ambulation, defecation, and response ambivalence behaviors were recorded in the one-way avoidance task in addition to response latency. WKY rats defecated more and produced lower scores on the three activity measures as compared to Wistar and F-344 rats. Response latency and ambivalence scores were higher for WKY rats. The subsequent ulcerogenic procedure also produced more ulcers in WKY rats. These data verify the ulcer susceptibility of WKY rats and underscore the assertion that their predominant stress coping behaviors are immobility and freezing. High ambivalence scores in WKY rats suggest behavioral inhibition, which is readily elicited by stressors. These behaviors resemble depressive behavior and suggest that WKY rat may be a useful animal model of depression. PMID- 8248373 TI - The effects of continuous morphine infusion on diet selection and body weight. AB - The administration of morphine causes a short-term increase in food intake, and repeated administration of morphine has been shown to cause progressively larger increases in intake and/or the relative intake of dietary fat. In this experiment, we measured the effects of continuous morphine infusions on diet choice and total intake. Male rats were given ad lib access to two diets: a high carbohydrate diet (80% carbohydrate, 20% protein) and a high-fat diet (80% fat, 20% protein). Diet intakes were measured daily for 21 days. Via the implantation of osmotic minipumps, one group received continuous infusions of morphine sulfate (approx. 2.8 mg/kg/h) for days 1-7 and of saline for days 8-14. A second group was infused with saline for days 1-7 and with morphine for days 8-14. A third group received sham surgery but no minipumps. Total caloric intake was significantly decreased on the final 6 days of morphine infusions. The percentage of total caloric intake consumed from the high-fat diet was significantly increased for the first 2-3 days of morphine treatment; this effect was due to an initial reduction in carbohydrate intake and an increase in fat intake. Over the course of the infusion period, fat intake gradually decreased and carbohydrate intake increased. The effects of morphine when infused on days 1-7 were similar to those observed when the drug was infused during days 8-14.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248374 TI - Maternal aggression of rats is impaired by cutaneous anesthesia of the ventral trunk, but not by nipple removal. AB - Aggression toward conspecific and allospecific individuals by female mammals is much more likely during lactation than during other reproductive states, a behavior that serves to protect the young. Previous research revealed that removal of nipples (thelectomy) prepartum greatly reduces the likelihood of postpartum aggression in house mice, but not in Sprague-Dawley Norway rats. The present work shows that prepartum thelectomy has no effect on the likelihood or intensity of postpartum aggression toward a strange male intruder in Long-Evans rats. In contrast, anesthesia of each nipple and surrounding skin prevents or severely impairs the elicitation of biting and attacking by the intruder, but does not impair normal retrieval of pups. Following removal of the litter, maternal aggression occurs readily at 1 h and somewhat less so at 5 h, but is absent at 24 h. These data suggest that while maternal aggression in postpartum rats does not require suckling, it is dependent on somatosensory stimulation of the ventral trunk by pups; this stimulation apparently produces a motivational change that lasts several hours. PMID- 8248375 TI - Laboratory instrumentation and computing: comparison of six methods for the determination of the period of circadian rhythms. AB - Data sets with known periodicity were used to compare the accuracy and noise tolerance of six methods of circadian period analysis: Fourier analysis, autocorrelation, Enright's (chi-square) periodogram, linear regression of onsets, interonset averaging, and acrophase counting. All methods except acrophase counting accurately detected the period of rhythms with pure waveform (cosine and square wave), whereas Fourier analysis and Enright's periodogram were superior to the other methods in the analysis of more complex waveforms (which more closely resemble actual circadian rhythms). The sensitivity of all methods was reduced by the insertion of random noise into the original data sets, but the methods of autocorrelation and Enright's periodogram were more tolerant of low signal-to noise ratios than the remaining methods. Although particular situations may require particular methods, the results suggest that Enright's periodogram is the best choice as a general method for the determination of the period of circadian rhythms. PMID- 8248376 TI - Effects of neuropeptide Y on ingestion of flavored solutions in nondeprived rats. AB - Recent evidence suggests that in addition to altering energy balance, neuropeptide Y (NPY) may stimulate ingestive behavior by modifying the orosensory quality of ingested substances. The present experiments investigated the effect of intracerebroventricular administration of NPY (5 micrograms/5 microliters) on ingestion of various flavored solutions in nondeprived rats. Experiment 1 examined the effects of NPY on ingestion of a range of concentrations of saline, sucrose, and saccharin solutions in single-bottle tests. Results indicated that NPY stimulates ingestion of both sucrose and saccharin solutions that are normally palatable. In Experiment 2, palatable sucrose solutions flavored with either orange or black cherry Kool-Aid for separate training groups were selectively associated with NPY injection during single-bottle training sessions. Subsequent two-bottle preference tests showed a significant shift in preference toward the flavor paired with NPY during training. The results of these experiments extend previous findings by showing that NPY can stimulate ingestion of sweet solutions regardless of caloric value and may potentiate sweet taste preference via an associative mechanism. PMID- 8248377 TI - Acute blockade of gastric emptying and meal size in rats. AB - To investigate the functional capacity of the combination of pregastric and gastric satiating stimuli for the control of meal size, rats were surgically implanted with a pyloric cuff that could be inflated to prevent gastric emptying. After 0.5 or 19 h of food deprivation, rats were given access for 30 min to three concentrations of sucrose (0.8, 0.4, and 0.2 M) and of corn oil emulsions (100%, 50%, and 25%) in descending order. There were only two significant changes in intake between cuff-open and cuff-closed tests and both occurred after 19 h of food deprivation: 1) meal size of 50% corn oil with the cuff closed was significantly larger than meal size of 50% corn oil with the cuff open on the preceding day; 2) meal size of 0.8 M sucrose with the cuff closed was significantly larger than meal size with the cuff open on the next test day. These results provide new experimental evidence that when ingested food is prevented from emptying from the stomach acutely, the satiating effect of food stimuli acting at pregastric and gastric sites is sufficient to produce meals of normal size under many, but not all, experimental conditions. PMID- 8248378 TI - Effects of 12-h sleep deprivation and of 12-h cold exposure on sleep regulation and cortical temperature in the rat. AB - The predictions of the two-process model of sleep regulation were tested by 12-h sleep deprivation (SDEP) and 12-h cold exposure (cold, 4 degrees C) in the rat, both carried out in the 12-h dark period. The analysis was based on recordings of vigilance states, electroencephalogram (EEG) power spectra (0.25-25.0 Hz), and cortical temperature (Tcrt). During recovery from SDEP, slow-wave activity (SWA; EEG power density in the 0.75-4.0 Hz band) in non-REM sleep was higher and the number of brief awakenings (nBA) lower than in the corresponding baseline period. During recovery from cold, a moderate and delayed increase in SWA was present. During SDEP, Tcrt was above baseline, and in some intervals of recovery below baseline. In those intervals waking was also decreased. During recovery from cold, Tcrt and waking were not affected. It is concluded that 12-h SDEP causes an intensification of sleep, as indicated by the enhanced SWA and the reduced nBA, whereas 12-h cold has only marginal effects. The time course of SWA in both experiments could be accurately predicted by a computer simulation based on the assumption of the two-process model and the parameter values determined in a previous experiment. PMID- 8248379 TI - Chronic vascular catheterization in the mouse. AB - A method is described for stress-free periodic sampling of blood and/or intravenous infusions in the normal awake and behaving mouse. The surgical procedure consists of the permanent implantation of a Silastic-rubber catheter in the right external jugular vein. The free end of the catheter is then pulled subcutaneously up to the skull and fixed on the skull with a form of dental acrylic that sticks firmly on the bone and thus does not require anchor screws in the skull. This technique allowed us to perform glucose tolerance tests in the mouse, and to follow the time course of blood glucose levels in individuals. PMID- 8248380 TI - Genetic differences in female house mice in aggressive response to sex steroid hormone treatment. AB - Male mice, genetically selected for aggression, characterized by short attack latency (SAL) or long attack latency (LAL), differ on several testosterone (T) related parameters during ontogeny and adult age. The variation in aggressive behavior at adult age may be due to differences in degree of androgenization prenatally. When exposed to T at prenatal, neonatal, and/or adult age, nonlactating females also display intraspecific fighting behavior. In the present study, we investigated in females of the SAL and LAL selection lines, whether the differentiation of aggression involves processes similar to ones seen in males. Therefore, we injected females with testosterone propionate (TP) or vehicle on the day of birth, treated them after ovariectomy at adult age with T, estradiol (E), or vehicle, and tested their aggressive response. We found that neonatally vehicle-treated SAL females show a higher aggressive response to chronic T treatment at adult age than LAL females receiving the same treatment. Females of both selection lines treated with vehicle or E as adults were not aggressive. Neonatal TP treatment did not influence the adult T sensitivity and difference between selection lines in response to T at adult age. However, neonatally TP treated SAL females showed aggressive behavior when treated with E at adult age, whereas LAL females failed to do so. These results suggest a genetic difference in susceptibility to T and E, which plays a major role prenatally, in organizing the development of sex steroid-dependent neural systems. PMID- 8248381 TI - Visual dysfunction in the spontaneously hypertensive rat. AB - We report two forms of visual defect in the spontaneously hypertensive rat (SHR) compared to its normotensive control strain (WKY). Ten-month-old male SHR and WKY were tested for intensity discrimination ability using a Y-maze and food reward. The SHR took more trials in the acquisition phase of the task using white light. In the test phase, in which the white light intensity difference was reduced, there was no significant difference in performance between SHR and WKY. Also, there was no significant difference between the strains when red light was used. The defect in SHR occurred in the blue range of the spectrum. Using blue light, the performance of SHR declined significantly at an intensity difference of 1.15 lux, whereas the performance of WKY did not decline significantly until the difference reached 0.01 lux. The defect in SHR for intensity discrimination was not related to cataract formation, but we detected a strain difference in the nuclei of the photoreceptor cells. Thus, this visual dysfunction may involve the rod photoreceptors. Further testing revealed a deficit in visual acuity in SHR aged 40-66 days, prior to the development of hypertension. Thus, the visual dysfunction of SHR is not secondary to their development of hypertension, but rather a discrete characteristic of the strain. PMID- 8248382 TI - Microstructural analysis of successive negative contrast in free-feeding and deprived rats. AB - Rats shifted from 1.0 M to 0.1 M sucrose lick at lower rates for the weaker solution than rats that have continual access to the 0.1 M sucrose solution only. This effect, referred to as successive negative contrast, has been investigated primarily in food-deprived rats and, in all cases, using total licks or total volume consumed as the dependent measure. The present experiment used a microstructural analysis of licking patterns to examine the changes in behavior that constitute the contrast effect in total licks in both deprived and free feeding rats. Although the magnitude of the effect was similar, deprived rats recovered from contrast more rapidly than free-feeding rats. Furthermore, the patterns of licking behavior associated with contrast differed under the two deprivation conditions. Specifically, when compared with the unshifted controls, the contrast effect in deprived rats was accomplished through a decrease in the number of licks per burst, an increase in the number of bursts initiated, a brief increase in the length of the interburst intervals, and no change in length of the interlick intervals. In free-feeding animals, contrast was associated with a decrease in the number of licks per burst, a brief increase in the length of the interburst interval, and no changes in either the number of bursts initiated or in the length of the interlick intervals. Together, these data demonstrate that patterns of licking behavior are differentially affected by solution concentration, deprivation state, and relative aspects of reward value. PMID- 8248383 TI - Voluntary intake and gastric emptying in pigs: effects of fat and a CCK inhibitor. AB - The influence of the cholecystokinin (CCK) antagonist MK-329 (0.1 mg/kg) on short term control of voluntary food intake and gastric emptying was studied in five pigs. Arterial injection of MK-329 abolished the inhibition of intake resulting from duodenal infusion of emulsified fat (Intralipid, 24 g/h). However, MK-329 only reduced the inhibition of dry matter (DM) and liquid emptying, and hence increased stomach volume at satiety. Intralipid infusion decreased the rise in gastric pressure during feeding. All these changes are consistent with endogenous CCK, released by Intralipid, inhibiting intake independent of the concomitant slowing of gastric emptying. MK-329 alone significantly increased intake using two different diets, but had no effect on gastric emptying so that gastric volume at satiety was significantly increased. These results suggest that endogenous CCK may be part of short-term satiety control in the pig. PMID- 8248384 TI - Sodium diclofenac inhibits hyperthermia induced by paradoxical sleep deprivation: the possible participation of prostaglandins. AB - Sodium diclofenac inhibits hyperthermia induced by paradoxical sleep deprivation (PSD), which suggests the participation of prostaglandins. The temperature of paradoxical sleep-deprived rats increased from the first to the fourth day of deprivation. This hyperthermia was blocked on the second, third, and fourth days by daily administration, twice a day, of 10 mg/kg of sodium diclofenac, a potent prostaglandin synthesis inhibitor. In the dose of 10 mg/kg, a decrease of temperature was observed only on the second and third days of PSD. These data suggest the participation of prostaglandins in modulating the increase in temperature during PSD. PMID- 8248385 TI - Lack of involvement of 4-hydroxynorephedrine in phenylpropanolamine-induced anorexia in rats. AB - The anorexic effects of phenylpropanolamine (PPA) appear to be qualitatively different in humans and rats. One factor that may account for these differences is that PPA is excreted essentially unchanged in humans, while nearly 30% is metabolized into 4-hydroxynorephedrine (4-OHN) in rats. To investigate the contribution of 4-OHN to the anorexic properties of PPA, this experiment compared the effects of equal doses (0.0-20.0 mg/kg, IP) of both drugs on eating and drinking during restricted feeding trials in the same group of food-deprived, female rats. Both 15.0 and 20.0 mg/kg of PPA significantly decreased eating when compared to saline vehicle, while 5.0-20.0 mg/kg of the drug reduced prandial drinking. In comparison, only the highest dose of 4-OHN (20.0 mg/kg) significantly suppressed food and water intake. When the percentage of reduction produced by corresponding doses of the two drugs was compared, PPA proved to be more than twice as potent as 4-OHN. It is concluded that, at the doses used, 4 OHN is unlikely to significantly contribute to reductions in deprivation-induced eating produced by the acute administration of PPA. PMID- 8248386 TI - Effect of early experience on behavior and immune response in the rat. AB - The effect of maternal deprivation (MD) and preweaning handling on open field (OF) behavior, body and organ weights (spleen, thymus, and adrenals), and humoral immune response (plaque-forming cell response and antibody production) in adult male and female Wistar rats was studied. Maternal deprivation took place either for 28 postnatal days (2 h/day), or on days 15, 18, and 21 (2 h/day), whereas handling was performed daily during 28 postnatal days for 3 min. Sex differences were found both in behavior and immune response. The MD rats showed ambulatory hyperactivity in OF tests, females being more active than males, and a marked suppression of the PFC response. Handled rat's behavior was distinguishable from MD rats by an increased curiosity. Female handled rats were more active in the OF and their antibody production was higher. Male handled rats showed higher defecation scores and lower plaque-forming cell response. These results present evidence for a deprivation syndrome and immunosuppressive behavior in MD rats. Several mechanisms that may account for these immunobehavioral results are outlined. PMID- 8248387 TI - A chronic high-salt diet fails to enhance blood pressure reactivity to a tone associated with footshock in SHR, BHR, and WKY rats. AB - Both the human and animal literatures suggest that reactivity to stress is enhanced in the presence of a positive family history of hypertension. There is also some suggestive evidence, though not as strong, that a high-salt diet will enhance reactivity to stress, at least in a subpopulation of individuals. In the present study, rats with zero (Wistar-Kyoto, or WKY), one (borderline hypertensive, or BHR), or two (spontaneously hypertensive, or SHR) hypertensive parents were placed on a normal or high (8% in chow)-salt diet for 8 weeks starting at 8 weeks of age. After 6 weeks on the appropriate diet, rats were stressed daily for 5 days. Each session consisted of 28 foot shock trials preceded by a tone. On the following week, animals were instrumented with femoral artery catheters. After a 2-day recovery period, they were again subjected to the experimental paradigm, during which blood pressure was continuously monitored. Differences were found with respect to blood pressure reactivity and family history: SHR were the most reactive to the tone associated with foot shock. However, no effects of salt on reactivity were observed, despite an effect of this manipulation on basal blood pressure. The effect of foot shock itself was also studied, and revealed that BHR showed a blood pressure reactivity response intermediate between SHR and WKY. Once again, no effects of the salt manipulation were seen. In conclusion, while the data support a relationship between family history of hypertension and reactivity to stress, they do not support a relationship between salt intake and reactivity to stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248388 TI - Neuroendocrine and behavioral effects of vasopressin in resting and mild stress conditions. AB - Neuroendocrine and behavioral effects of subcutaneously administered AVP (6 micrograms/kg b.wt.) were determined in resting conditions and after the mild stress of transportation to and placement in a novel environment. In resting conditions, systemic administration of AVP caused a rapid increase in blood glucose level and a long-lasting increase in CORT secretion. A decrease in circulating plasma NE level was observed. AVP did not affect adrenal medullary E secretion. The stress-induced sympathetic activation, as reflected in plasma NE level, was inhibited 60 min after AVP administration. Stress-induced increases in blood glucose, CORT, and E secretion were not influenced by AVP. In resting condition, AVP caused a 60-min lasting increase in grooming behavior with a concomitant decrease in time spent resting. Sixty minutes after administration, the behavioral activation after the stress of transportation and placement in a novel environment was more rapidly terminated in AVP-treated rats than in vehicle treated control rats. The state- and time-dependent modulation of spontaneous behavior suggests that AVP has arousing properties shortly after administration when marked pressor and endocrine effects are apparent. In a later phase, AVP may facilitate dearousal mechanisms following stress-induced behavioral activation. PMID- 8248389 TI - Induced components in the spontaneous ovulatory cycle of the Djungarian hamster (Phodopus campbelli). AB - Although Djungarian hamsters, Phodopus campbelli, show regular, 4-day spontaneous estrous cycles when tested for behavioral receptivity, this study shows that over 50% of 624 virgin females paired with males show their first behavioral estrous on the third day after pairing. The observed pattern is compared against predictions of estrous synchrony and/or estrous cycle suppression models. They fail to explain the high incidence of mating on the third day, and are incompatible with our current understanding of the natural history of the species. An alternative hypothesis, that Djungarian hamsters fail to cycle in the absence of a male, but are rapidly induced into a succession of 4-day cycles by his presence, appears to explain both the pattern of matings and earlier observations that estrous cycles could not be monitored by vaginal smear or lavage, only by interactions with a male. These analyses, thus, conclude that Djungarian hamster females may require a signal from a male to initiate and maintain successive 4-day estrous cycles. PMID- 8248390 TI - Does a learned taste aversion contribute to the anorectic effect of bacterial lipopolysaccharide? AB - The present study addressed the possible role of a conditioned taste aversion in the anorectic effect of bacterial lipopolysaccharide (LPS) in the rat. Pairing an intraperitoneal (IP) injection of LPS (100 micrograms/kg b.wt.) with the subsequent presentation of a familiar diet (FD) or of a novel-tasting saccharin diet (SD) for several hours did not affect FD or SD intake when the same diet was offered several days later after 12 h of food deprivation. However, food intake during the second presentation of SD was reduced when food was not withheld prior to the test. In a similarly designed experiment, the antipyretic and antiinflammatory drug indomethacin (5 mg/kg b.wt., IP) attenuated the anorectic effect of LPS during the initial pairing, but did not affect the inhibition of SD intake in LPS-pretreated rats during the second feeding test. The antiemetic trimethobenzamide (5 mg/kg b.wt., IP) failed to influence the anorectic effect of LPS. Lesion of the area postrema (AP) and the adjacent nucleus of the solitary tract (NST) was found to enhance the anorectic effect of LPS, but the development of tolerance to this effect remained unchanged in AP/NST-lesioned animals. In spite of the ability of LPS to induce a taste aversion that inhibits feeding under certain conditions (novel-tasting diet, no food deprivation prior to the feeding test), the findings indicate that a learned taste aversion is not the only contributor to the anorectic effect of LPS. PMID- 8248391 TI - Effects of sleep deprivation, lunch, and personality on performance, mood, and cardiovascular function. AB - The present study examined the effects of sleep deprivation on performance, mood, and cardiovascular functioning in the late morning and early afternoon. The results showed that the sleep-deprived subjects felt less alert and detected fewer targets in a cognitive vigilance task. Selective impairments due to sleep deprivation were also observed in a logical reasoning task. There was little evidence to suggest that consumption of lunch altered these effects of sleep deprivation. However, individual differences in the effects of sleep deprivation were apparent, with trait anxiety being related to the changes in subjective alertness produced by sleep deprivation, and sleep-deprived extraverts showing greater performance impairments than the sleep-deprived introverts. PMID- 8248392 TI - Ethophysiological responses to overwinter food shortage in wild European rabbits. AB - Wild European rabbits studied under seminatural conditions turned out to have about 30% lower heart rates during periods of overwinter food shortage than during periods with access to green forage or pelleted rabbit chow. Concomitant changes in the animals' metabolic rate (ADMR), body weight, blood sugar level, and foraging activity clearly indicate that this heart rate reduction is a sign of starvation due to insufficient food quality. Telemetered body temperature, on the other hand, was amazingly stable in most subjects, even during overwinter food shortage. These data are discussed with respect to nutritional requirements and possible mechanisms of energy conservation in wild European rabbits. PMID- 8248393 TI - Interactions among aging, gender, and gonadectomy effects upon naloxone hypophagia in rats. AB - The present study examined the dose-dependent (0.25-5 mg/kg) effects of systemic naloxone upon deprivation-induced intake and high-fat intake as functions of age (4, 8, 14, and 20 months), gender, and gonadectomy in rats. Significant increases in body weight were observed as functions of age and gonadectomy. Whereas aging significantly reduced basal deprivation-induced intake, it generally failed to alter basal high-fat intake. Whereas age, gender, and gonadectomy failed to alter the decreases in deprivation-induced intake following low (0.25-2.5 mg/kg) naloxone doses, sham males displayed significantly greater age-related and gender related inhibition following the 5 mg/kg dose of naloxone. Young gonadectomized rats displayed significant increases in naloxone's inhibition of deprivation induced intake as well. More dramatic changes occurred in naloxone's inhibition of high-fat intake. Naloxone's potency increased in sham female rats as a function of age, and decreased in sham males and ovariectomized females as a function of age. Whereas sham males and ovariectomized females were most sensitive to naloxone's inhibition of high-fat intake at young ages, sham females were most sensitive at older ages. These data indicate that effects of age, gender, and gonadectomy upon naloxone-induced hypophagia dissociate as a function of the type of intake. Because selective opioid antagonist studies demonstrate that deprivation-induced intake is mediated by the mu1 receptor and high-fat intake is mediated by kappa and mu2 receptors, it is postulated that the differential effects of aging, gender, and gonadectomy variables upon opioid mediation of the two forms of intake may reflect their interaction with different opioid receptor subtypes. PMID- 8248394 TI - Amygdaloid NMDA and muscarinic receptors involved in working memory performance of rats. AB - In a three-panel runway task, the competitive N-methyl-D-aspartate (NMDA) receptor antagonist, 3-[(+-)-2-carboxypiperazin-4-yl]propyl-1-phosphonic acid (CPP) (56 ng/side), injected bilaterally into the basolateral subdivision of the amygdala, significantly increased the number of errors (attempts to pass through two incorrect panels of the three panel gates at four choice points) in a test of working memory. Intra-amygdaloid injection of the muscarinic receptor antagonist scopolamine (5.6 micrograms/side) also significantly increased working memory errors. In a test of reference memory, neither CPP nor scopolamine affected the number of errors when injected into the basolateral amygdala at doses up to 56 ng/side or 5.6 micrograms/side, respectively. These results indicate that processes mediated by NMDA and muscarinic receptors in the basolateral amygdala are involved in the performance of working memory, but not in reference memory. PMID- 8248395 TI - Learned preferences for the flavor of salted food. AB - We examined whether rats can associate the flavor of their food with its salt content, and whether this association is influenced by sodium status during training and testing. During two pairs of 2-h training trials, rats ate flavored food containing 1.75% NaCl or an alternatively flavored unsalted food. The motivational state of the rats was manipulated prior to each trial by combined 48 h dietary sodium deprivation and furosemide treatment (severe sodium depletion), 48-h dietary sodium deprivation (mild sodium deprivation), or continued maintenance on stock diet containing 1% NaCl (sodium replete). When later given a choice between the two flavors, all rats preferred food containing the salt paired flavor. The strength of this preference was unaffected by motivational state during training or by the salt content of the test foods, but was modulated by the motivational state of the rats during the preference test. Preference for the salt-paired flavor was strongest when rats were tested after severe sodium depletion, less strong after mild sodium deprivation, and absent when sodium replete. These results indicate that deprivation state during training has little effect on learned preferences for the flavor of salted food but deprivation state during testing affects the expression of this learning. PMID- 8248396 TI - Management of locally aggressive sinus disease using craniofacial exposure and the galeal frontalis fascia-muscle flap. AB - Histologically, benign sinus diseases can behave aggressively, potentially causing sight- and life-threatening conditions requiring surgical treatment. Four patients illustrating acute and chronic infection, single and multiple sinus polyposis, and mucoceles are presented and discussed to illustrate how the principles of craniofacial exposure, resection, and reconstruction may be employed to treat aggressive sinus disease. The bicoronal and buccal sulcus approaches minimize visible scarring; wide exposure of the orbital contents and dura reduce the risk of excessive bleeding, neurologic complications, and orbital volume derangements. Immediate cranial bone grafting and the well-vascularized galeal-frontalis myofascial flap obliterate the dead space and isolate the extradural space from contaminated facial cavities, diminishing the risk of infection. The craniofacial surgical approach is particularly appropriate for the management of aggressive frontoethmoidal and multiple sinus involvement because it allows the treatment of sinus cavities under direct vision and complete resection of sinus disease. PMID- 8248397 TI - Comparison of CT imaging artifacts from craniomaxillofacial internal fixation devices. AB - This study compares the artifacts caused by craniomaxillofacial internal fixation devices in CT images. Mandibular reconstruction and "mini" titanium, Vitallium, and stainless steel systems, "micro" titanium and Vitallium systems, and stainless steel wires were evaluated. The hardware was placed on a nylon grid and submerged in water. CT images were obtained with both bone and soft-tissue window settings. All artifacts were compared and graded after a minimum of five observations each. The severity of "starburst" artifact was found to be related to the physical size of the fixation hardware and its composition. Titanium hardware caused the least amount of artifact. Vitallium and stainless steel fixation devices, with the exception of interfragmentary wiring, produced significantly more artifact. These results agree with theoretical predictions. The data indicate that when postoperative imaging is an important clinical consideration, (1) the least amount of implant material necessary to achieve stable fixation should be used, (2) the proximity of implant material to the area of interest should be considered, and (3) titanium implants produce less artifact than Vitallium or stainless steel implants. PMID- 8248398 TI - Clinical experience with the tunable pulsed-dye laser (585 nm) in the treatment of capillary vascular malformations. AB - A retrospective review of 134 patients who received laser therapy with the pulsed dye laser at the 585-nm wavelength for capillary vascular malformations was conducted. These patients were treated over a 5-year period with a minimum follow up of 2 years. Ninety-five patients were available for evaluation of results. Thirty-nine were lost to follow-up. Each birthmark was described before and after treatment under a classification system developed by the authors. Results were reported by classification, age, anatomic location, and race. Excellent or good results were obtained in the majority of patients in all age groups and regardless of the color of the birthmark. All anatomic areas and all ethnic groups responded to treatment with varying degrees of success. A range of up to eight treatments was required to achieve resolution. Scarring was an infrequent complication (1.1 percent) of treatment. Hypopigmentation was seen in 3.2 percent of treated patients. The pulsed-dye laser (585 nm) offers promise to patients of all ages and hope to those who previously were not candidates for laser therapy. PMID- 8248399 TI - Repositioning the lateral alar crus. AB - Malposition of the lateral crus is a normal variant of the nasal alar cartilage as described by Sheen which shows a "parentheses" deformity from the frontal view. The cephalic-positioned lateral crus leaves the alar rims without cartilaginous support, causing deformity and potential destabilization of normal airway competence. The procedure described herein is a modification of the Sheen technique that can be done as either an open or a closed procedure. It is particularly valuable in decreasing the overprojecting nasal tip and preserving the equal height of the alar domes while widening the angle of divergence of the lateral crus. This technique delivers consistently dependable results for an anatomic variant modification frequently overlooked in rhinoplasty surgery. PMID- 8248400 TI - Surgical anatomy of the SMAS: a reinvestigation. AB - There has been considerable disagreement in descriptions of the SMAS. Fresh cadaver dissection, combining anatomic dissection done with the aid of the operating microscope and histologic cross section of facial tissues, was performed in 24 facial halves. Histologic preparation was made on tissue macrosections, each 10 cm in length. Consistent findings include the following: (1) Although the SMAS is closely applied to the superficial surface of the parotid gland, a thin but distinct parotid fascia can be identified between parotid gland and SMAS. (2) The SMAS terminates superiorly within 1 cm below the zygomatic arch, and is not continuous with the temporoparietal fascia. (3) The SMAS in the cheek lies in continuity with the orbicularis oculi muscle of the lower eyelid. (4) Considerable variability exists in the histologic appearance of the SMAS in different facial regions within a single cadaver, as well as in a given facial region between cadavers. Because of its variability, histologic identification of the SMAS should be made in continuity with known reference structures, such as the platysma. PMID- 8248401 TI - Classification of mandibular defects. AB - A classification of mandibular defects based on functional as well as aesthetic factors is presented. By taking into account the difficulties in restoring form and function and not simply relying on traditional anatomic landmarks, it is hoped that this method will allow different types of reconstructions to be fairly evaluated. It also should help surgeons to tailor individual reconstructive techniques to specific clinical situations. Major difficulties in mandibular reconstruction arise when a condyle requires replacement, when there is a mucosal and/or skin component to the defect, and when the area to be reconstructed involves the anterior arch. The classification is based on three upper-case and three lower-case characters: H, C, L and o, m, s. H defects are lateral defects of any length, including the condyle but not significantly crossing the midline; L defects are the same only without the condyle; C defects consist of the entire central segment containing the four incisors and the two canines. Combinations of these letters are possible (an angle-to-angle defect, for example, is represented as LCL). Thus H and L defects may reach or even extend slightly beyond the midline but are not referred to as LC or HC unless they contain the entire central segment. The letters o (neither a skin nor a mucosal component), s (skin), m (mucosa), and sm (skin plus mucosa) are added to denote the epithelial requirement. PMID- 8248402 TI - Reduction mammaplasty by suction alone. AB - Because unsatisfactory scars often result when traditional methods of reduction mammaplasty are performed, techniques that leave less noticeable scars have been advocated recently. One such technique is suction lipectomy, which has been employed previously, but only for patients who have well-located, small nipple areola complexes and whose hypertrophy is primarily fat. I have found that concern for the size and location of the nipple-areola complex and cause of hypertrophy are unfounded. Thus reduction mammaplasty by suction alone may be the technique of choice in certain patients. Eleven patients had reduction mammaplasty by suction alone and obtained aesthetically pleasing breasts that had good sensation and minimal scars. As a result, the method is suggested for selected patients: those with minimal to moderately large breasts. PMID- 8248403 TI - Mammary ducts in the areola: implications for patients undergoing reconstructive surgery of the breast. AB - The use of selective excision of the nipple with preservation of the areola ("nipple coring") has been advocated for patients undergoing subcutaneous or total glandular mastectomy. This is based on the assumption that all the mammary ducts in the nipple-areola complex are located in the nipple and that the areola is devoid of mammary ductal tissue. To investigate this issue, we performed nipple coring on eight consecutive mastectomy specimens, excised the areolas, and submitted them in their entirety for histologic examination. Mammary ducts identical in appearance to extralobular ducts in the breast parenchyma were identified in the dermis of the areola in all eight cases. These ducts were present throughout the areola, including the most peripheral aspects. We conclude that mammary ducts are a normal component of the areolar dermis and that nipple coring does not result in the removal of all mammary ductal tissue from the nipple-areola complex. This observation should be taken into consideration by reconstructive surgeons considering preservation of the areola in patients undergoing subcutaneous or total glandular mastectomy. PMID- 8248404 TI - The permeability of tissue expanders to bacteria: an experimental study. AB - Infection is an occasional but serious complication of tissue expansion. Bacteria in many cases are found both inside and outside the expander. The mechanism of how they spread inside and out is not fully understood. In the current study, we have assessed both the expander membranes (n = 28) and the port (n = 42) in terms of leakage and transmission of Staphylococcus aureus in normal expansion (100 percent) and overexpansion (400 percent) in an in vitro model. Our results indicate that the membrane was impermeable to S. aureus, whereas the bacteria were able to migrate freely through the puncture sites in the ports. PMID- 8248405 TI - Blood conservation in massive suction lipectomy. AB - The practice of massive liposuction for the treatment of patients with large lipodystrophies has already been reported. The safety of this technique lies in proper fluid and blood replacement. Blood conservation techniques are therefore needed to avoid or minimize homologous transfusion. The use of a combination of predeposited autologous blood and acute intraoperative normovolemic hemodilution in the management of 150 patients requiring massive liposuction is reported. Since it is possible to anticipate the amount of blood loss in liposuction, a formula is proposed to monitor the hematocrit of the patient during the procedure and therefore predict the need for homologous transfusion. The technique reported enabled us to aspirate up to 11 liters in a single stage. Homologous transfusion was either avoided or markedly reduced. The degree of patient satisfaction was high, with no mortality or morbidity reported. PMID- 8248406 TI - The effect of systemic antibiotic and antibiotic-impregnated polymethylmethacrylate beads on the bacterial clearance in wounds containing contaminated dead bone. AB - A dorsal muscular wound model was used in 40 New Zealand White rabbits to study the effect of systemic and local antibiotics on the bacterial clearance of contaminated dead bone. Devitalized iliac crest bone preincubated with Staphylococcus aureus was implanted in each deep muscular wound with or without tobramycin-impregnated polymethylmethacrylate beads. Either systemic tobramycin or cefazolin was administered for 7 days. Animals were sacrificed at 7 and 14 days. The wounds containing tobramycin beads had significantly fewer bacteria than those without antibiotic beads (2.0 x 10(2) versus 1.3 x 10(6); p < 0.008). The reduction in bacteria due to the tobramycin beads did not differ significantly with respect to the concurrent systemic antibiotics or to the duration of incubation. We conclude that tobramycin-impregnated beads are effective in reducing bacterial count in contaminated bony wounds treated with systemic antibiotics. Furthermore, the bactericidal effect of the antibiotic beads is independent of and additive to the systemic antibiotic delivered to the wounds by well-perfused muscles. PMID- 8248407 TI - One-stage total penile reconstruction with a free sensate osteocutaneous fibula flap. AB - This is the first series of total penile reconstructions with the free sensate osteocutaneous fibula flap. The main advantages of this flap lie in its intrinsic rigidity, its superior donor-site location, and its long vascular pedicle. The fibula flap provides better bone volume than does the radial forearm flap, which commonly results in a floppy phallus in the absence of bone. Penile prostheses in other flaps have enjoyed limited success. Forearm donor-site complications can be avoided. The donor site in the lower extremity can be readily covered with a sock. The vascular pedicle of the fibula flap is of sufficient length to allow end-to-side anastomosis of the flap to the femoral artery. Interpositional vein grafts are unnecessary, and dissection of the inferior epigastric artery system to serve as a donor artery may be avoided. The appearance of the neophallus is excellent. We present only the first four continuous cases of the six we have performed because sufficient follow-up data are available only for these four. The advantages and disadvantages of fibula and forearm donor sites, the long-term fate of the bony component, the importance of sensation, and the vascularized urethral reconstruction are discussed. High patient satisfaction and the advantages of the technique convince us that the fibula osteocutaneous flap is superior for total penile reconstruction. PMID- 8248408 TI - Dorsalis pedis flap with vascularized extensor tendons for dorsal hand reconstruction. AB - We describe three cases of combined loss of skin and tendons on the dorsum of the hand treated with the use of cutaneotendinous dorsalis pedis free flap. The functional and aesthetic results are excellent in the hand and aesthetically acceptable in the foot. This technique is the first choice when the treatment of these injuries requires three or four tendon grafts. PMID- 8248409 TI - Sixty cases of partial or total toe transfer for repair of finger losses. AB - Our experience with partial or total toe transfers for the reconstruction of finger deficits is presented. Sixty-one toes were transferred to reconstruct finger deficits in 60 patients. The transfers from the big toe consisted of 3 vascularized whole-nail grafts, 4 onychocutaneous flaps which included the nail and a skin flap from the toetip, 16 thin osteo-onychocutaneous flaps, 4 wrap around flaps, and 3 combined wrap-around and dorsalis pedis flaps. The transfers from the second toe consisted of 6 trimmed toetips including the nail, 16 second toes, 1 combined second toe and a dorsalis pedis flap, and 1 second toe and third toe. The other transfers mainly consisted of other flaps, including a hemipulp flap and a first web space flap. Regarding the transfers from the big toe, vascularized nail grafts and onychocutaneous flaps were found to be most suitable for the treatment of total nail loss, thin osteo-onychocutaneous flaps for distal phalangeal loss of the thumb, wrap-around flaps with a vascularized iliac bone graft for thumb loss above the metacarpal joint, and the combined wrap-around and dorsalis pedis flaps for a total thumb deficit. Regarding the transfers from the second toe, the trimmed toetips including the nail were most suitable for claw nail deformities, the second toe was most suitable for finger loss except for the thumb with the proximal interphalangeal joint, and the combined second toe and dorsalis pedis flap was most applicable for a total thumb deficit including thenar skin loss. PMID- 8248410 TI - Diffusion versus iontophoresis in the transport of gentamicin in the burned rabbit ear model. AB - This experimental study demonstrates that iontophoresis can be used to transport biologically active gentamicin into auricular cartilage. Fifty female New Zealand White rabbits were divided into three groups: live controls with unburned ears, live animals with burn-injured ears (deep or partial-thickness), and euthanized animals with unburned ears. Each group was then divided into subgroups that received gentamicin transported by iontophoresis or diffusion. In ears subjected to iontophoresis, adequate gentamicin activity could be demonstrated only in the cartilage of euthanized animals with unburned ears and live animals with full thickness burned ears. Diffusion did not transport a detectable level of gentamicin in any instance. Diffusion is ineffective in the transport of gentamicin and should not be used as a primary method of treatment for ear chondritis. Iontophoresis did not transport active gentamicin across partial thickness burns, presumably because the migrating antibiotic molecules were dispersed throughout the body by the circulation. However, if the circulation is destroyed, as in a full-thickness burn, iontophoresis can move the biologically active gentamicin into the cartilage. PMID- 8248411 TI - Predicting the survival of experimental skin flaps with a knowledge of the vascular architecture. AB - Experimental skin flaps have been utilized by researchers for almost a century to investigate many of the perplexing questions in plastic and reconstructive surgery, yet the underlying vascular anatomy of these flaps is addressed infrequently. The purpose of this study was to predict the survival of experimental skin flaps prior to their elevation in guinea pigs and rabbits, planned on a knowledge of the underlying vascular anatomy. On the basis of our previous anatomic studies, 17 guinea pigs and 15 rabbits were used in separate experiments. In experiment 1, two parallel flank flaps of identical dimensions were compared on one side of each guinea pig. The dorsal flap encompassed the vascular territories of multiple perforators, while the ventral flap embraced two perforators (two-territory flap). Viability was assessed on days 3 and 7 by inspection and fluorescein dye injection. All ventral flaps survived to a greater extent than the dorsal flaps. Whole-body fresh cadaver lead oxide injections were performed to provide cutaneous angiograms. It was found in each flap that the area of skin viability corresponded to the capture of one to two adjacent vascular territories on the artery at its base. In the second experiment, a multiple-territory osteocutaneous flap was designed on one side of the torso of the rabbit using the Doppler probe. It was based on the thoracodorsal artery and embraced the skin and a 1 x 2 cm segment of iliac crest bone in the adjacent deep circumflex iliac artery angiosome. Using the same criteria as in experiment 1, we found that one to two adjacent viable skin territories were captured on the thoracodorsal artery. In addition, viability of the iliac bone was confirmed in every case by angiography, fluorochrome labeling, and india ink injection studies indicating the capture of deep structures of the deep circumflex iliac artery angiosome. This study reinforces the angiosome concept and indicates that one adjacent vascular territory may be captured reliably in experimental guinea pig and rabbit skin flaps. In addition, we have described a reliable osteocutaneous flap model in the rabbit. PMID- 8248412 TI - Plastic surgery board review courses: to review or to reveal ... that is the question. PMID- 8248413 TI - A dilemma. PMID- 8248414 TI - Rigid fixation, skull reconstruction, and fiscal responsibility. PMID- 8248415 TI - Midazolam and sexual fantasies. PMID- 8248416 TI - Extradigital glomus tumor as a cause of knee pain. AB - Although there exists much debate in the dermatohistopathologic literature about the exact characterization of the glomus tumor, this case illustrates what should be considered the sine qua non indications of clinically significant glomus tumors, i.e., exquisite pain and complete relief of pain with surgical excision. One should not, therefore, limit the inclusion of glomus tumors to the differential diagnosis of severe localized pain only in the upper extremity. PMID- 8248417 TI - Squamous cell carcinoma of the lacrimal sac. AB - Squamous cell carcinoma of the lacrimal sac is rare. We report a case of a 45 year-old Caucasian woman with a squamous cell carcinoma of the right lacrimal sac, initially presenting at an early stage. A critical review of the literature is also presented with specific regard to operative and postoperative management. Treatment of these lesions is predominantly surgical. The role of radiation therapy is not yet fully defined. Successful management requires a high index of suspicion as well as aggressive and early intervention. PMID- 8248418 TI - A new free thenar flap. AB - While previous pedicled procedures yielded good results in a small traumatic volar defect of the fingers, a large volar defect presents a difficult reconstructive challenge. We devised a new free thenar flap utilizing the superficial palmar branch of the radial artery which we used to treat two patients with volar defects of the index finger. This free flap provides an excellent alternative in coverage of a large long volar defect of the fingers with little sacrifice of the donor site. PMID- 8248419 TI - Postburn facial resurfacing with a split ascending neck flap. AB - Ninety-eight patients with postburn facial scar deformities were treated by using a split ascending neck flap. Anatomic studies and discovery of axial blood supply to the anterior of the neck are utilized, allowing for wide mobilization, thin flaps, and transfer of neck skin to the face and chest skin to the neck. By leaving the platysma muscle in situ and using vacuum drainage with a large tube in the subflap space, preservation of the chin and neck contours is obtained. There are five varieties of postburn facial scar deformities described and treated. Extensive facial deformity requires a two-stage operation. Neck skin transferred to the face by the described method preserves all natural properties. This method of facial resurfacing is preferable to other methods and is indicated for all cases of scars to the face with healthy neck skin and chest wall. PMID- 8248420 TI - Repair of a recurrent urethrovaginal fistula with an island bulbocavernous musculocutaneous flap. AB - This paper describes a case of recurrent postpartum urethrovaginal fistula. The extent of the vaginal tissue loss and the perilesional scarring made direct closure of the defect not practicable. After suturing of the urethra, the anterior vaginal wall was reconstructed with an island bulbocavernous musculocutaneous flap raised from the left labium majus. Seven months after surgery, the flap healed well, and cystography showed a regular voiding without periurethral suffusions. Healing of thedonor site also was aesthetically satisfactory. PMID- 8248421 TI - A study of the outcome of the American Society for Aesthetic Plastic Surgery research grant program. AB - Since 1974, the American Society for Aesthetic Plastic Surgery has funded a competitive research grant program jointly administered by the Plastic Surgery Educational Foundation. The grant program was initiated to provide "seed money" to researchers in aesthetic surgery. A written survey of all recipients over a 6 year period from 1985 to 1990 was conducted to assess the productivity of the grant program. The survey suggests that seed money grants are effective and that there is a high rate of peer-reviewed publication and national presentation of completed projects. Suggestions for improvement of the grant program are also presented. PMID- 8248422 TI - An instance of failure of local anesthesia during liposuction: cause? PMID- 8248423 TI - Injection of botulinum A toxin for hemifacial spasm. PMID- 8248425 TI - Medications that cause gynecomastia. PMID- 8248424 TI - The use of an island chondromucosal flap in eyelid reconstruction: preliminary report. PMID- 8248426 TI - 31P magnetic resonance spectroscopy to study noninvasively metabolism in cutaneous tissue. PMID- 8248427 TI - Augmentation of craniofacial skeleton. PMID- 8248428 TI - Lip enhancement. PMID- 8248429 TI - Autologous fat injection. PMID- 8248430 TI - Fixation of vascularized bone graft in mandibular reconstruction. PMID- 8248431 TI - Vermilion reconstruction with labia minora graft. PMID- 8248432 TI - An observation of the dental anlage based on findings in cleft lip patients. PMID- 8248433 TI - Tumescent technique for liposuction. PMID- 8248434 TI - Death after stapling of skin graft to intestine. PMID- 8248435 TI - Measurement of the excised skin in face lifting procedure. PMID- 8248436 TI - Preoperative columella lengthening in bilateral cleft lip and palate. PMID- 8248437 TI - Postoperative headaches following large-volume suction lipectomies. PMID- 8248438 TI - Calcification bonded to saline-filled breast implants. PMID- 8248439 TI - [Psychiatry and the law in German-speaking countries]. PMID- 8248440 TI - [Criteria for assessing the ability of psychiatric patients to give informed consent]. AB - In this paper criteria for the assessment of the ability of psychiatric patients to give an informed consent are presented. Legal and medical aspects including jurisdiction are discussed. Some elements for a judgement of an informed consent are proposed. PMID- 8248441 TI - [Alcohol and drug abuse as risk factors for violence and delinquency in schizophrenic patients]. AB - A study on the prevalence of substance abuse in schizophrenics performed in 1989/1990 had shown high rates of alcohol and drug abuse in schizophrenia. An additional analysis of the data revealed that schizophrenics with substance abuse had been convicted more often than other schizophrenics. The possible role of substance abuse as a risk factor for delinquency or casualties in schizophrenia is discussed. PMID- 8248442 TI - [Sentenced to psychotherapy? Report of experiences from the Austrian central treatment institution for incompetent mentally abnormal criminals]. AB - A short report is given of the legal principles underlying the treatment of mentally ill offenders in Austria-called the "Massnahmenvollzug". Pro and contra of psychotherapy under a compulsory setting in a specialized prison is discussed. Some concepts of inpatient psychotherapy are described and their realisation in "Justizanstalt Gollersdorf"-an Austrian prison for adult mentally abnormal offenders-is given in development and modifications for all day living. The discussion of theory and casework leads to some conclusions: Psychotherapy under the conditions of "Massnahmenvollzug" seems to be possible if mutually free decision for it can be guaranteed. The therapist must be willing to take care and responsibility not only for the psychotherapeutic process alone but for the structure of prison organisation as well. The institution as a whole should be organized under therapeutic conditions. PMID- 8248443 TI - [Discharge of incompetent mentally abnormal criminals: experience from the Austrian central treatment institution]. AB - Based on the data of 66 male patients not guilty for reason of insanity, who were treated in Justizanstalt Gollersdorf, Austria's central institution for mentally ill offenders, we examined the influence of age, duration of detention, diagnosis, and offence on discharge. Law provides a yearly, compulsory examination of the necessity of further inpatient-treatment. We also tried to describe a concept of further dangerousness (considered to be the essential basis for the decision of possible discharge) in a four-factor-model including the criteria psychopathology, insight, conflict behaviour, and social competence. Concerning the various levels of decision finding (institution, expert, court) different interpretations of these criteria could be demonstrated. Independent of diagnostic and prognostic considerations the court stressed mainly kind of offence as the most important factor for the possibility of discharge. PMID- 8248444 TI - [Social workers in the commitment procedure]. AB - Regular inclusion of a social worker alongside the expertising physician whenever a judge has to decide on committing a person to a psychiatric hospital, is known in German legal practice as "Stuttgart model". Our experiences with this procedure are discussed on an interdisciplinary level. In our opinion this model agrees with the spirit of the new German law governing guardianship and care. PMID- 8248445 TI - [Pedophilia and diaper fetishism in a man with Klinefelter syndrome]. PMID- 8248446 TI - ["Psychologic marital unfitness" in cyclothymia? A case report on the topic of the annullment procedure in canon law]. AB - According to the Canon Law (CIC) of the catholic church a marriage can be annulled because of psychological inability under certain conditions. We present the case report of a married woman, who fell ill with cyclothymia after her first childbirth. Her marriage was annulled under canon 1095 (n.3 CIC) because of psychological inability to fulfill martial obligations. The authors believe that this represents a clear cut case of discrimination against patients with mental disorders: on the one hand the catholic church claims that every martial bond is principally indissoluble; on the other hand they do not eschew invalidating a marriage due to an affective psychosis. This poses special problems which are also discussed. PMID- 8248447 TI - Guidelines for the recording and quantitative analysis of electroencephalographic activity in research contexts. AB - Developments in technologic and analytical procedures applied to the study of brain electrical activity have intensified interest in this modality as a means of examining brain function. The impact of these new developments on traditional methods of acquiring and analyzing electroencephalographic activity requires evaluation. Ultimately, the integration of the old with the new must result in an accepted standardized methodology to be used in these investigations. In this paper, basic procedures and recent developments involved in the recording and analysis of brain electrical activity are discussed and recommendations are made, with emphasis on psychophysiological applications of these procedures. PMID- 8248448 TI - Modulation of event-related potentials by word repetition: the role of visual selective attention. AB - Event-related potentials (ERPs) were recorded while subjects viewed visually presented words, some of which occurred twice. Each trial consisted of two colored letter strings, the requirement being to attend to and make a word/nonword discrimination for one of the strings. Attention was manipulated by color in Experiment 1, and color and a precue were used in Experiment 2. As in previous ERP studies of word repetition, a positive offset to repeated words developed when both first and second presentations were the focus of attention. In Experiment 2, ERPs showed evidence of positive-going repetition effects in all conditions in which at least one of the two presentations of the repeated word was attended. In the visual modality, the positive-going ERP repetition effect occurs only when at least one of the two presentations of a repeated item is the object of attention, which suggests that one or more of the processes reflected by the effect is capacity limited. PMID- 8248449 TI - Perceptual context and the selective attention effect on auditory event-related brain potentials. AB - Two experiments examined the effect of the perceptual context established through tonal grouping on neuroelectric responses during selective listening. Subjects monitored one of the extreme pitches in four-pitch tone sequences and detected rare longer tones of the designated pitch. In the first study, tonal grouping was manipulated by changing the tonal separation between the extreme pitches and their nearest neighbor, keeping the extreme pitches constant. Grouping increased the negativity of the attended-unattended difference potential. A second study examined the effect of grouping on the attention-related negativity when it opposed the effect of physical similarity. The proximity of extreme pitches varied, keeping the middle pitches constant. The effect of grouping on event related potentials (ERPs) for the middle pitches was varied according to the tone being attended. ERPs for the middle tone adjacent to the attended tone became more negative with grouping, whereas ERPs for the distant middle tone became less negative with grouping. These effects suggest that the attention-related negativity is sensitive to contextual information. PMID- 8248450 TI - Startle reflex facilitation as a function of classical eyeblink conditioning in humans. AB - I investigated whether classical eyeblink conditioning increases the activating effect of the conditional stimulus (CS) on the relevant alpha motoneuron pool in the facial nucleus. The activating effect was measured by the reflex modification method, where startle reflexes, sharing the facial nerve motor pathway with conditional and unconditional eyeblink reflexes, were elicited in the presence of reinforced and nonreinforced CSs (CS+ and CS-). To reduce influences of conditional arousal on startle, a weak airpuff to the eye was used as unconditional stimulus and relatively short stimulus onset asynchronies (SOAs) of 300 and 600 ms were used in three startle tests presented while conditioning was in progress. Two groups (N = 72) received 100% or 0% reinforcement (Groups 100 and 0) of a tone CS+. The CS- was not reinforced. Three startle tests, consisting of acoustic startle probes presented alone and after CS+ and CS- onset, were conducted early, middle, and late in conditioning. Results showed increased startle reflex facilitation after CS+ onset in the late startle test in Group 100, indicating that startle increased as a function of classical conditioning. Although reflex facilitation at the 300-ms SOA in Group 100 is consistent with the hypothesis that CS presentations activated motoneurons in the facial nucleus, the present study cannot itself definitively support this hypothesis. Reflex facilitation at the 600-ms SOA in Group 100 could be influenced by conditional arousal. PMID- 8248451 TI - Amplitude and bilateral coherency of facial and jaw-elevator EMG activity as an index of effort during a two-choice serial reaction task. AB - In earlier studies, positive but inconsistent relationships have been reported between mental effort and electromyogram (EMG) amplitude in task-irrelevant limb muscles. In this study, we explored whether facial EMG activity would provide more consistent results. Tonic EMG activity of six different facial and jaw elevator muscles was bilaterally recorded during a two-choice serial reaction task with paced presentation of auditory or visual signals. In Experiment 1, task load (signal presentation rate) was kept constant for 20 min at the level of the subject's maximal capacity. In Experiment 2, task load was increased in a stepwise fashion over six successive 2-min periods from sub- to supramaximal capacity levels. EMG amplitude and coherency between momentary bilateral amplitude fluctuations were measured. In Experiment 1, EMG amplitude of frontalis, corrugator supercilii, and orbicularis oris inferior showed a strong gradual increase throughout the task period, whereas taks performance remained fairly stable. Orbicularis oculi, zygomaticus major, and temporalis EMG showed a much smaller increase or no increase. In Experiment 2, the first three muscles showed a fairly consistent increase in EMG amplitude with increasing task load. Orbicularis oculi and zygomaticus major were not active until task load became supramaximal. Effects of stimulus modality or laterality were not found in any experiment. These results are consistent with the notion that EMG amplitude of frontalis, corrugator, and orbicularis oris provides a sensitive index of the degree of exerted mental effort. PMID- 8248452 TI - Autonomic activity in boys with disruptive behavior disorders. AB - Skin conductance (SC) and heart rate (HR) were measured during rest, a series of tones, and a reaction time task in 34 boys with disruptive behavior disorder to ascertain (a) if this broadly defined group differed from control boys (n = 33) and (b) if there were differences within the spectrum related to component diagnoses or to target behaviors. Disruptive boys had higher resting HR than controls, due largely to those without a subdiagnosis of conduct disorder. Disruptive boys showed a smaller increment in SC response frequency for task instructions. They did not show a general deficit in phasic SC reactivity but habituated to signal stimuli at a faster rate than did controls. However, their SC response latencies were shorter despite slower motor reaction time. Responsivity variables were similar in boys with and without a subdiagnosis of conduct disorder. The data are not compatible with the hypothesis of low autonomic nervous system (ANS) baselines in disruptive boys but partially suggest low ANS reactivity. They generally support the validity of the disruptive behavior disorder spectrum. Correlations with measures of aggression and impulsivity suggest that individual differences within the spectrum might be more fruitfully described by behavior dimensions than by conventional subdiagnosis. PMID- 8248453 TI - The relationship between trait hostility and cardiovascular reactivity: a quantitative review and analysis. AB - Results of a series of meta-analyses indicated that high and low scorers on most trait hostility measures do not consistently differ in blood pressure or heart rate reactivity to traditional laboratory stressors. The few significant effects were modest in size, and instances of hyporeactivity were found. When stressors were classified as provocative versus nonprovocative, in accord with Trait x Situation approaches, however, Potential for Hostility-Interpersonal Style was predictive of exaggerated systolic and diastolic blood pressure responses and the Cook-Medley Hostility Inventory was predictive of diastolic blood pressure responses to provocative stressors. Hence, the next generation of studies of the hostility-reactivity hypothesis should emphasize interpersonal stressors. Alternative mechanisms for the disease consequences of hostility should also be examined, however, because the available evidence indicates that the hyperreactivity hypothesis is unlikely to furnish a complete explanation for the association between hostility and heart disease. PMID- 8248454 TI - Reliable responses to a cardiovascular reactivity protocol: a replication study in a biracial female sample. AB - This replication study was designed to examine the reliability of individual differences in cardiovascular responsivity to a standardized PC-based cognitive task protocol (Kamarck et al., 1992) in a female community sample. Thirty women, both black and white (ages 25-44 years), were administered the protocol twice with a 1-month retest interval using a mobile laboratory at a community-based testing site. Measures of heart rate and blood pressure reactivity were obtained comparing assessments taken at rest and during three protocol tasks. Preejection period, stroke volume, and total peripheral resistance changes were estimated using impedance cardiography assessments. As in previous studies with males, individual differences in heart rate and systolic and diastolic blood pressure responses to the protocol were highly reliable (.80 or greater) when data were aggregated across three tasks and two testing sessions. Reliability of cardiac contractility (preejection period) and stroke volume changes to these tasks exceeded .70. This reactivity assessment procedure has now produced reliable results in three studies and appears to be exportable across diverse samples and settings. PMID- 8248455 TI - Familial influences on heart rate, blood pressure, and self-report anxiety responses to stress: results from 100 twin pairs. AB - Two hundred healthy adolescent to middle-aged individuals (12-44 years, M = 20 years) were tested in a standardized stress protocol. These individuals comprised 20 monozygotic female, 20 monozygotic male, 20 same-sex dizygotic female, 20 same sex dizygotic male, and 20 opposite-sex dizygotic twin pairs. Familial influences on heart rate, blood pressure, and self-report anxiety responses to four different kinds of stressors (Visual-Verbal Test for Conceptual Thought, mental arithmetic, isometric handgrip, cold pressor) were assessed using biometrical genetic model fitting. Evidence of significant genetic effects on resting heart rate and blood pressure was obtained, providing heritability estimates of .65, .63, and .58 for resting heart rate and systolic and diastolic blood pressure, respectively. Cardiovascular reactivity to the Visual-Verbal Test, mental arithmetic, and the cold pressor test appeared to be primarily influenced by genetic and idiosyncratic (nonfamilial) environmental factors, whereas reactivity to handgrip was more related to effects of the family environment. The results of multivariate model fitting suggested that the genetic effects on reactivity were relatively independent of those affecting resting heart rate and blood pressure and that there was significant overlap of genetic influences on heart rate and blood pressure responses to the two active coping tasks. PMID- 8248456 TI - Memory template comparison processes in anhedonia and dysthymia. AB - Anhedonic subjects, potentially at risk for psychopathology because of a deficient ability to experience pleasure, have demonstrated a large N200 component in the event-related brain potential (ERP). The present experiment attempted to determine the psychological significance of this finding in light of Naatanen's (1990) distinction between N2a and N2b subcomponents. Anhedonics were contrasted with controls and dysthymics, an at-risk group reporting depression. Across groups, N2a was larger when a tone mismatched a longer run of preceding identical tones. Thus, an involuntary mismatch process appears to be intact in both at-risk groups. However, the three groups produced distinct N2bs as a function of stimulus sequence. The N2b finding for anhedonics is consistent with Knight's (1984, 1992) model of early stimulus processing deficits in schizophrenia. PMID- 8248457 TI - A method of constant stimuli for examining heartbeat detection: comparison with the Brener-Kluvitse and Whitehead methods. AB - A method of constant stimuli (MCS) for measuring heartbeat detection was examined by comparing performance on this method with the Brener and Kluvitse (BK) method and a variant of the Whitehead (WH') method. All methods required subjects to judge the simultaneity of heartbeat sensations and tones, and each yielded measures of judgment precision and of the temporal location of heartbeat sensations relative to the electrocardiogram R-wave. Both measures were found to be significantly correlated across tasks. A greater proportion of subjects met criteria for classification as heartbeat detectors on the MCS (54%) and BK (50%) tasks than on the WH' task (33%). In the MCS and BK tasks, subjects judged tones presented 100-300 ms after the R-wave to be most simultaneous with heartbeat sensations. Intratask correlations showed that only the MCS procedure yielded stable measurements of both judgment precision and temporal location. The MCS procedure possesses commendable psychometric properties and provides a simpler means than the BK procedure of examining heartbeat detection. PMID- 8248458 TI - The relationship between neuroticism and blood pressure reexamined. An investigation of a nonclinical sample of military conscripts. AB - Given the contradictory results reported in the literature on the relationship between neuroticism and blood pressure, it was decided to reexamine this topic. This was done with a large sample very homogeneous for age, sex, food intake and life style and basically unselected in terms of medical history. In the frame of a blood donation program blood pressure was measured in 810 conscripts. In addition, subjects completed either form A or form B of the Eysenck Personality Inventory (EPI). Data from a total of 624 subjects were suitable for evaluation. No significant correlations were obtained between systolic or diastolic blood pressure and scores of neuroticism or extraversion (r = -0.10 to 0.06) nor did groups with different blood pressure values differ substantially in terms of these variables. This supports work showing that there is no essential connection between blood pressure and emotional lability as measured by the EPI. PMID- 8248459 TI - Relaxation therapy in the treatment of bronchial asthma: effects on basal spirometric values. AB - Eighteen asthmatic children were divided into two 9-patient groups matched with respect to severity of the disease. The first group received a 6-week asthma self management programme which included progressive relaxation training. The second group received the same programme without the relaxation training component. State anxiety and spirometric data (FEV1, FVC and FEV1/FVC%) for both groups, obtained before and after each session with the therapist, showed that relaxation is effective in reducing state anxiety but does not modify basal spirometric values. PMID- 8248460 TI - Technical advances in behavioral psychotherapy of obsessive-compulsive disorders. PMID- 8248461 TI - Aspects of abnormal illness behaviour. AB - This paper reviews in detail the concept of abnormal illness behaviour. Recent developments in relation to criteria for diagnosis are discussed. Newer approaches to management are described, with particular attention to controlled trials. The need for a 'transition phase' of therapy is emphasized. PMID- 8248462 TI - Hypnosis and relaxation in the medically ill. AB - Interest in the application of hypnotic techniques for patients with medical disorders seems to rise and fall over the years. Enthusiasm for this work comes both from patients and clinicians. Often, however, these techniques are offered without regard to the psychological theories that should inform their operation and the limits that clinical and experimental research suggest. This article offers a brief description of the elements of hypnosis and a review of the history of the use of hypnotic techniques with a variety of medical problems including asthma, habits such as cigarette smoking and medical symptoms such as persistent nausea and vomiting. Special attention is placed on the psychological and physiological principles that help to establish the valid use of this technique. PMID- 8248464 TI - A curious irony. PMID- 8248463 TI - Are parents of asthmatic children different from other parents? Some follow-up results from the Norwegian PRAD Project. AB - Follow-up results from the Norwegian PRAD study (Psychosocial Risk for Allergic Development) show that parents of children who later developed symptoms of asthma were different from a control group of parents with normal, nonsymptomatic children several years preceding the disorder. There were significant within-pair differences in self-reported marital adjustment (DAS-Dyadic Adjustment Scale) for the control group but not for the asthma group. There were no significant between pair DAS scores for the two groups of parents. The two groups of parents also differed in perceiving how the child had influenced their lives, in description of the childbirth and partially in childcare style. The assumed relationship between parental characteristics and asthma in children is being discussed, and the paper concludes that parents of asthmatic children should be regarded as a risk factor and clinical and therapeutic routines should be developed accordingly. PMID- 8248465 TI - The influence of psychosocial factors on survival after myocardial infarction. AB - The prognostic importance psychosocial factors after acute myocardial infarction (AMI) is still debated. A fourfold increase in risk of mortality after AMI was reported for participants in the beta Blocker Heart Attack Trial who were described as being socially isolated and having high levels of life stress. This study was designed to determine the influence of social isolation and/or life stress on mortality after AMI in an English population. It was a follow-up study of a subset of patients recruited for the Anglo-Scandinavian Study of Early Thrombolysis (ASSET), between November 1986 and February 1988. The study group comprised 1,376 patients with suspected AMI (1,073 men and 303 women), from coronary care units in six English hospitals. Patients who were alive at 7 days and had completed a psychosocial questionnaire within seven days post-infarction were followed up for a median time of three years, the sole outcome measure being death from all causes. All deaths were notified by the National Health Service Central Registry (NHSCR). Cox's regression was used to allow for independent clinical prognostic factors such as age-group, previous documented infarct, complications in hospital, history of diabetes and history of hypertension. Socially isolated patients (in terms of lack of membership of any club or religious group or lack of contact with family and friends) were 49% more likely to die after an infarction than patients classified as not being socially isolated. While this statistic is of borderline significance it does suggest that such patients are at an increased risk of death after AMI. No associations with mortality risk were found with life-stress level, type A behaviour pattern or depression. PMID- 8248466 TI - The four levels of health promotion: an integrated approach. AB - Four levels of health promotion are identified: environmental, social, organisational and individual. It is argued that health promotion interventions should not be confined to one level but instead an integrated approach should be adopted, in which the relationships between the four levels and the outcomes at all levels ought to be considered and analysed. It is suggested that these four levels are used as a checklist when health promotion interventions are planned. PMID- 8248467 TI - The QALY: a guide for the public health physician. AB - This paper considers the background to the rational allocation of health resources at DHA level within the context of the new purchaser/provider regime. A popular approach to this problem has been that of cost utility analysis, using quality adjusted life years (QALYs) as the measure of the benefit derived from a health intervention. This paper seeks to offer the practising public health physician an appraisal of the practical value and pitfalls of a QALY-based approach. To this end, the theoretical background to QALYs and their practical use are described and some of the practical and ethical problems and pitfalls are discussed. It is concluded that the QALY concept provides a useful tool in aiding decision making in the allocation of health care resources, but that it is too early for a universal application of the approach. PMID- 8248468 TI - Costs to affected individuals following an outbreak of food poisoning: a pilot study. AB - This small pilot study surveyed the victims of an outbreak of food poisoning, in order to investigate the personal consequences which they suffered so as to identify costs, which are often ignored in economic analyses. A response rate of 88% was achieved. The results showed that an average of 10.5 days of illness was suffered by the affected individuals and there was a marked variation in the number of days of work missed. Housewives were identified as a group whose personal consequences are often omitted from economic studies and a variety of other costs were identified. The results should be borne in mind when undertaking future economic studies of food poisoning. PMID- 8248469 TI - Room for improvement? Detecting autosomal trisomies without serum screening. AB - OBJECTIVE: To assess the efficacy of the prenatal screening and diagnostic programme for the detection of the common autosomal trisomies in the North West Thames region prior to the introduction of serum screening. DESIGN: A retrospective study based on reports made to the North West Thames Regional Health Authority Congenital Malformation Register. SUBJECTS: 235 pregnancies of women delivered in units in the North West Thames region over a two-year period (1990-91) whose babies or fetuses were diagnosed as having Down, Edwards or Patau syndrome. RESULTS: 33% of Down syndrome, 68% of Edwards syndrome and 52% of Patau syndrome were diagnosed prenatally (before 28 weeks) in the region without the use of serum screening. CONCLUSIONS: Using maternal age and routine ultrasound as screening methods, units in the region are prenatally diagnosing significantly fewer Down syndrome babies than are those in authorities using serum screening. The use of maternal age and ultrasound screening is more effective for the antenatal detection of the other autosomal trisomies than it is for the detection of Down syndrome. It is not yet clear to what extent serum screening might be expected to increase the detection rate for these conditions. PMID- 8248470 TI - Hospital bed usage by people with HIV disease: experience in a provincial setting. AB - In order to describe hospital bed usage by people with HIV disease in a provincial setting, a retrospective analysis of admissions to hospital wards in Oxford was undertaken for people admitted to hospital with all HIV-related illnesses or complications of HIV-related treatment. A total of 83 people were identified as having been admitted to hospital between January 1986 and the end of August 1990. Average length of hospital stay, the number of admissions per observed person-year and the in-patient days per observed person-year decreased. Of the 2,446 days spent in hospital, 913 were by people with an AIDS diagnosis; 1,533 days were spent by people who did not fulfil the World Health Organisation/Centers for Disease Control (WHO/CDC) classification for AIDS but who were admitted because of their HIV disease. AIDS is an end-point of infection with HIV. Pre-AIDS morbidity, a spectrum of illness of increasing severity from minor illness up to the point of WHO/CDC level AIDS, is a major determinant of hospital care and has previously been underestimated. In order to calculate the best estimates of hospital care needed, HIV disease should be regarded as a single entity and the artificial barrier dividing HIV illnesses from AIDS should be discarded. PMID- 8248471 TI - Community needlestick accident data and trends in environmental quality. AB - New health-based indicators of environmental quality are being sought. In this search a retrospective analysis was undertaken for all persons in England and Wales who, during 1988-91, were given Hepatitis B Immune Globulin (HBIG) in prophylaxis against Hepatitis B infection after accidental percutaneous or permucosal exposure, and for whom a surveillance form was returned to the Public Health Laboratory Service Communicable Disease Surveillance Centre, England and Wales (PHLS CDSC). Although the number of adult HBIG doses issued during 1988-90 increased annually from 1,907 to 2,850, the percentage for which completed forms were returned fell during this time from 55% (1,043/1,907) to 31% (894/2,850). In 1991, 2,049 doses were issued and completed forms returned for 32% (653/2,049). From data on the completed and returned forms for 1988-91, 51% (1,805/3,535) of accidental exposures occurred in the community and 45% (1,604/3,535) in hospitals. Of the community exposures, 958 (53%) were needlestick accidents; 16% of them occurred 'in the street', 12% from 'contact with rubbish', 6% 'in the park', and 4% 'on the beach'. The findings confirm that discarded hypodermic syringe needles in the general environment are causing new public health problems. As the percentage of completed and returned forms has declined, and because HBIG is not recommended for needlestick accidents in the community where the source of the needle is unknown, HBIG surveillance forms are thought not to be a suitable source of data for continued monitoring of environmental quality. PMID- 8248472 TI - Calculation of photon mass energy-transfer and mass energy-absorption coefficients. AB - Calculations of mass energy-transfer and mass energy-absorption coefficients for photon energies from 1 keV to 100 MeV have been developed, based on a re examination of the processes involved after the initial photon interaction. The probabilities for the initial interaction are from the current photon interaction cross-section database at the National Institute of Standards and Technology. The calculations then take into account (1) electron binding effects on the Compton scattered photon distribution; (2) the complete cascade of fluorescence emission after ionization events in any atomic subshell, including those associated with incoherent scattering and triplet production; and (3) the radiative energy losses of the secondary electrons and positrons slowing down in the medium, including the emission of bremsstrahlung, characteristic X rays from impact ionization, and positron in-flight as well as at-rest annihilation quanta. Consideration of the processes in (3) goes beyond the continuous-slowing-down approximation and includes the effects of energy-loss straggling. Results for the mass energy absorption coefficient are compared with those from recent tabulations. PMID- 8248473 TI - Characterization of the products of dinucleoside monophosphates d(GpN) irradiated in aqueous solutions. AB - The dinucleoside monophosphates d(GpT), d(GpC), and d(GpA) were X-irradiated in oxygenated aqueous solution. The principal products were identified in the intact modified dinucleoside monophosphates using NMR spectroscopy and FAB mass spectrometry. The 8-hydroxyguanine modification is a major product in each of the d(GpN). The d(GpN), where N is a pyrimidine nucleoside, also yield products in which the pyrimidine base is degraded to a formamido remnant. The most interesting product is one bearing two base damages, 8-hydroxylation of the guanine base and degradation of the pyrimidine base to a formamido remnant. This double lesion was observed in both d(GpN) where N is a pyrimidine base. PMID- 8248474 TI - Bone sarcoma characteristics and distribution in beagles fed strontium-90. AB - A total of 66 primary bone sarcomas were diagnosed in 47 beagles; 43 of these dogs were part of the 403 beagles fed 90Sr and 4 were part of the 162 controls. Multiple primary bone sarcomas were found in 15 of the 47 beagles (32%). The incidence of multiple primary bone sarcoma was restricted to the two highest dose groups, except for a single control dog which developed two bone sarcomas. A threshold-like radiation dose response was observed; no sarcomas were observed in the lowest three dose groups, but the number of primary bone sarcomas increased rapidly in the higher dose groups. Of the 66 primary sarcomas, 49 were osteosarcomas (74%). As the dose increased, the proportion of osteosarcomas increased sharply, 4/10 (40%), 26/29 (90%), and 16/18 (89%), in the three highest dose groups. Thirteen of the bone sarcomas of other types occurred in males, and 4 in females, whereas 21 osteosarcomas occurred in males, and 28 in females. The ratio of bone sarcomas of the appendicular skeleton to those in the axial skeleton was 40:26, with osteosarcomas occurring more often in the appendicular than the axial skeleton (32:17), whereas nonosteogenic tumors showed no predilection (8:9). A statistical study of the distribution of bone sarcomas among 16 separate bone groups showed a correlation only with the distribution of cancellous bone volume-to-surface ratio and not with either skeletal mass distribution or dose distribution. The highest occurrence of sarcomas was in the humeri, femora, and mandible, and no occurrence in the coccygeal vertebrae, paws, or sternum. It is postulated that the distribution of bone sarcomas reflects a critical combination of the osteosarcoma precursor cell population, their cell division rate, and the radiation dose absorbed by these cells. PMID- 8248475 TI - Yield of strand breaks as a function of scavenger concentration and LET for SV40 irradiated with 4He ions. AB - We have measured by gel electrophoresis the yields of single- and double-strand breaks (SSBs and DSBs) induced in aqueous solutions of SV40 DNA and the SV40 minichromosome by 137Cs gamma rays (mean LET 0.3 keV micron-1) and 4He ions (mean LETs 85, 102, and 152 keV microns-1). DNA SSBs are caused mainly by the hydroxyl radicals under these conditions and are reduced in yield as either the hydroxyl radical scavenger concentration or the LET is increased (over the range studied). The G(SSB) for 4He ion irradiation is less by a factor of up to 10 than the G(SSB) for gamma irradiation, depending upon the scavenger concentration. The difference in the yields of SSBs agrees well with the difference in the yields of hydroxyl radicals for the radiations in question. In contrast, the yields of DSBs are similar for gamma and 4He ion irradiation over much of the range of scavenging capacity studied. However, at the highest scavenger concentrations the yields of DSBs are greater for 4He ion irradiation. In addition, the yields of DSBs remain almost constant with increasing LET (over the range studied). Therefore the relative yield of DSBs per SSB increases with increasing LET, supporting the hypothesis that increasing LET leads to an increased clustering of damage in DNA. PMID- 8248476 TI - Increased DNA repair ability after irradiation following treatment with the immunomodulator AS101. AB - Ammonium trichloro(dioxoethylene-o-o')tellurate (AS101) is a new synthetic compound previously described by us as being able to modulate the immune system and having minimal toxicity. Clinical trials are currently in progress with AS101 on cancer patients. AS101 has recently been found to have radioprotective effects on hemopoiesis and survival of irradiated mice when administered prior to irradiation. Radioprotection conferred by AS101 has recently been demonstrated by us to result partly from induction of progenitor cells to enter into S phase, which is assumed to be a more radioresistant phase of the cell cycle, and partly from the enhanced stimulation of CFU-S not only toward proliferation but also toward self-renewal. In the present study we demonstrate that the DNA repair processes expressing the cellular reponses associated with the restoration of the normal nucleotide sequence after damage caused to the DNA were also increased significantly after treatment with AS101. Unscheduled DNA repair synthesis was found to be significantly higher in both spleen and bone marrow cells from mice injected with AS101 compared to mice injected with PBS. DNA repair synthesis in spleen cells incubated with AS101 in vitro was also higher than that of PBS treated cells. This was demonstrated by equilibrium alkaline cesium chloride density gradient of DNA from irradiated and nonirradiated spleen cells in the presence of hydroxyurea. In addition, using the neutral filter elution technique, we show that AS101 can both protect cells from DNA double-strand breaks (DSBs) induced by irradiation and enhance the ability of the affected cells to rejoin the DSBs. We show that extracts of splenocytes, either incubated with AS101 in vitro or obtained from mice injected with AS101, contain substantial DNA polymerase activity which is significantly higher compared to that of control treated cells. Aphidicolin, an inhibitor of DNA polymerases alpha and delta, and dideoxy-thymidine, an inhibitor of DNA polymerase beta, inhibited DNA repair synthesis of irradiated splenocytes stimulated with AS101. These results collectively indicate that AS101 confers its radioprotective effects partly by preventing the induction of DSBs induced by irradiation and partly by enhancing the ability of irradiated cells to repair their damaged DNA, probably by increasing mainly DNA polymerase activity. The understanding of the mechanism of radioprotection conferred by AS101 will enable us to use AS101 more effectively for the restoration of hemopoiesis in patients after radiation therapy or in patients suffering from overdose or accidental irradiation. PMID- 8248477 TI - Radiation hepatology of the rat: parenchymal and nonparenchymal cell injury. AB - Cellular changes that occur within the liver before and during liver fibrosis have been studied. At various times (0-90 days) after gamma irradiation (0-25 Gy) of the whole liver, hepatocytes and liver nonparenchymal cells were isolated by enzymatic dissociation of the liver and differential centrifugation. Differential cell counts were done to quantify the yield of hepatocytes and nonparenchymal cells. Hepatocyte function in vitro was assayed by the uptake of rose bengal and compared with the clearance of the dye in vivo. Intracellular alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were used to estimate the effect of radiation on the phenotypic expression of hepatocytes. Gene expression of transforming growth factor beta 1 (TGF-beta 1), a cytokine which has been found to play a role in hepatic fibrosis, was measured by 32P-cDNA:mRNA hybridization of poly(A+) mRNA isolated from purified populations of hepatocytes and nonparenchymal cells. Morphologically, hemorrhages were evident in perfused livers 28 days after 25 Gy whole-liver irradiation. This injury was preceded by a significant increase in liver nonparenchymal cells at 2 weeks, the earliest time after irradiation at which measurements were made. Significant increases in liver nonparenchymal cells occurred at doses greater than 10 Gy and persisted throughout the follow-up period after irradiation. The lowest dose studied (5 Gy) inhibited both normal growth of the liver and the associated increase in the number of hepatocytes. Doses greater than 10 Gy resulted in a dose-dependent decline in liver mass and hepatocytes. Viable hepatocytes isolated from irradiated dysfunctional livers, with respect to clearance of rose bengal, were functionally normal in vitro. A dose-dependent decline in the ALT and AST contents of liver cell suspensions and homogenates was observed but the relative decline in ALT was greater, resulting in lower ALT/AST ratios. This decline in the ALT/AST ratios occurred at doses (< 15 Gy) which caused no loss of hepatocytes, suggesting possible conversion of ALT-rich periportal hepatocytes to ALT-poor hepatocytes due to changes in the microenvironment and/or aging of the cell population. TGF-beta 1 mRNA was detected mainly in nonparenchymal cells, and radiation preferentially enhanced the TGF-beta 1 gene expression in these cells. These data suggest that radiation-induced alterations in liver nonparenchymal cell populations may be responsible for microvascular fibrosis, which results in a cascade of pathological events which lead to hepatocyte loss. PMID- 8248478 TI - Radiation hepatology of the rat: time-dependent recovery. AB - In situ recovery kinetics of gamma-irradiated rat livers was investigated. To measure short-term repair, surgically exposed livers were irradiated with a 12.5 Gy priming dose. At various times (0-24 h) the liver was re-exposed surgically to 10 Gy. To measure long-term recovery, rats were exposed to a 10-Gy priming dose to the upper abdomen to irradiate the whole liver. The liver was then re-exposed surgically and given graded test doses 0 to 56 days after the first exposure. Repair as a function of interfraction interval (1-12 h) between eight fractions of 3 Gy to the upper abdomen followed 2 weeks later by an 10-Gy top-up dose to the exposed liver was also investigated. Approximately 3 months after the last exposure plasma aspartate aminotransferase, retention of intravenously injected rose bengal, liver weight, and hydroxyproline content were measured to determine hepatic necrosis, function, mass, and fibrosis, respectively. A 15-Gy single exposure threshold dose was needed to produce detectable hepatic injury. Exceeding this threshold dose resulted in a dose-dependent increase in liver injury as measured by all four end points. Split-dose irradiation with a 10-Gy priming dose and an interfraction interval of 1 day increased the isoeffect dose by 3.9 to 5.8 Gy. Increasing the interfraction interval from 1 to 56 days did not further increase the isoeffect dose. Exposing the liver to eight fractions of 3 Gy with an interfraction interval of 3 h or longer resulted in a dose recovery of approximately 80% (i.e., 19 Gy). A substantial portion of this recovery was evident with interfraction intervals of 1 h. A fluctuating repair pattern with increasing time (0 to 24 h) between 12.5 and 10.0 Gy liver irradiation was observed. This repair was characterized by a 1-h delay after irradiation, followed by near complete repair during the next 3 h, followed by sensitization during the next 2 h and a second wave of repair between 8 and 24 h. Repair was not completed until 21 h after irradiation. It is concluded that the liver's capacity to repair injury within 24 h is similar to that of other late-responding tissues. However, there is no additional repair after 24 h, indicating an absence of slow repair. PMID- 8248479 TI - Long-term observation of the rat skin after fractionated local X irradiation: development of secondary tumors. AB - Data from previous animal experiments were analyzed retrospectively with a view to the induction of secondary tumors. The skin of 86 rats was exposed locally to multiple fractions of 200 kV p X rays at doses of 60 to 82 Gy applied in 30 or 35 fractions in 6 weeks, i.e., in the clinically relevant range of 1.9 to 2.7 Gy per fraction. The course of the early skin reaction was scored and compared to the late carcinogenic effects of the treatment using Kaplan-Meier data analysis. With increasing total dose the median duration of the early skin reaction increased, while median latency of tumor induction in the irradiated area of skin decreased from 381 days after 60 Gy to 274 days after 82 Gy. Independent of the total dose, number of fractions, and operative intervention, 84 to 100% of the animals cured by irradiation developed a secondary neoplasm inside the treated area. The majority of induced tumors were squamous cell carcinomas. In the skin covering the thorax, which had received a dose of 15 Gy in 10 fractions for elective irradiation of the lungs, no tumors were induced. PMID- 8248480 TI - Stimulation of hematopoietic cell recovery by tetrachlorodecaoxide in sublethally irradiated mice. AB - This study investigated whether tetrachlorodecaoxide (TCDO), a potent wound healing agent, promotes recovery of hematopoietic tissue of mice depleted by sublethal X irradiation. Mice were exposed to 3 Gy total-body irradiation and were given 1 ml/kg TCDO intravenously daily from day 1 to 5 or day 4 to 8 after irradiation. The effect was assessed by the change in the total number of nucleated cells in bone marrow and spleen, endogenous spleen colony formation, and 30-day mouse lethality. In the spleen there were profound effects on spleen weight, total organ cellularity, and endogenous colony formation. The TCDO produced radiation dose-modifying factors between 1.4 and 1.5 for endogenous spleen colony formation in mice primed with 3 Gy and reirradiated 4 days after treatment. This stimulated cellular recovery resulted in an increased protection of mice from lethality caused by subsequent total-body irradiation by factors of 1.12-1.18. The results show that TCDO is a potent stimulator of cellular recovery in the spleen of sublethally irradiated mice. PMID- 8248481 TI - Cytogenetic response to G2-phase X irradiation in relation to DNA repair and radiosensitivity in a cancer-prone family with Li-Fraumeni syndrome. AB - Noncancerous skin fibroblasts from six family members with Li-Fraumeni syndrome, five with cancer of diverse tissue origin and one with a premalignant neoplasm, showed a high frequency of chromatid aberrations, 94 to 119 breaks and 58 to 95 gaps per 100 metaphase cells arrested with colcemid 0.5 to 1.5 h after X irradiation (1.75 x 10(-2) C/kg). This response results from deficient repair of the radiation-induced DNA damage. In contrast, skin fibroblasts from two unrelated normal controls and a spouse showed 19 breaks and 17 to 19 gaps per 100 cells. Whereas all six members of the cancer-prone family had a radioresistant phenotype, only four had an inherited p53 mutation. Fibroblasts from a radioresistant family member showed the same extent of chromatid damage directly (0 to 0.5 h) after G2-phase X irradiation as those from the radiosensitive control spouse. We conclude, therefore, that radiosensitivity, as determined by cell killing in asynchronous populations of skin fibroblasts, is unrelated to chromosomal sensitivity to G2-phase X irradiation. However, the persistence of a high frequency of chromatid breaks and gaps at 0.5 to 1.5 h after G2-phase X irradiation, a manifestation of deficient DNA repair, is associated with proneness to cancer in this family. PMID- 8248482 TI - An ionizing radiation-sensitive mutant of CHO cells: irs-20. II. Dose-rate effects and cellular recovery processes. AB - An ionizing radiation-sensitive mutant of CHO cells, irs-20, irradiated in plateau-phase cultures showed a complete lack of a dose-rate effect, i.e., there was no reduction in cell killing per unit dose as the dose rate was lowered from 0.75 Gy/min to 0.06 Gy/h. No measurable repair of potentially lethal damage or sublethal damage was evident for irs-20 cells in the G1 phase of the cell cycle. During S phase, irs-20 cells showed repair of sublethal damage similar to that for the wild-type cells. PMID- 8248483 TI - Gamma radiation (5-10 Gy) impairs neuronal function in the guinea pig hippocampus. AB - Guinea pigs were exposed to 5 and 10 Gy gamma radiation. Hippocampal brain slices were isolated 30 min, 1 day, 3 days and 5 days after irradiation or sham irradiation and the electrophysiological characteristics of the neural tissue were evaluated. Both radiation doses elicited significant changes that were dependent on dose, dose rate and time. Synaptic efficacy decreased soon after exposure to 5 Gy at dose rates of both 1 and 20 Gy/min. Recovery occurred by 5 days. Ten grays at 20 Gy/min potentiated the postsynaptic potential 1 day after irradiation. By 3 days, synaptic efficacy was decreased and did not recover. The ability of the synaptic potentials to generate spikes was potentiated within 30 min after exposure to 5 Gy at 1 Gy/min and persisted through 3 days, with recovery at 5 days. At the 20 Gy/min dose rate, a similar potentiation did not result with 10 Gy and occurred only at 3 days after irradiation with 5 Gy. Rather, within 30 min and after 5 days, spike generation was significantly depressed by these exposures. Both synaptic efficacy and spike generation contribute to the net input-output relationship of the neuronal population. This relationship was profoundly decreased within 30 min with recovery at 1 day and subsequent decline with the higher dose rate in a dose-dependent manner. These persistent changes in neuronal function are likely to be a consequence of the actions of ionizing radiation on the physiological processes that influence the neuronal environment. PMID- 8248484 TI - Ionizing radiation induces two forms of interphase chromosome breaks in Chinese hamster ovary cells that rejoin with different kinetics and show different sensitivity to treatment in hypertonic medium or beta-araA. AB - We have shown previously that incubation of irradiated plateau-phase CHO cells in hypertonic growth medium during the period normally allowed for chromosome condensation, in the premature chromosome condensation (PCC) assay, uncovers a form of interphase chromosome breaks that rejoin with fast kinetics (t1/2 = 1.5 min). Here, we report that incubation with beta-arabinofuranosyladenine (beta araA), an inhibitor of DNA, chromosome, and cellular repair processes, during the same period uncovers a different form of interphase chromosome breaks that rejoin with slower kinetics (t1/2 = longer than 15-20 min). The yield of interphase chromosome breaks increased from 2.0 breaks/cell/Gy in untreated control cells to 3.6 breaks/cell/Gy in cells exposed to 1 mM beta-araA, and was the same as that observed in cells treated in hypertonic medium (500 mM NaCl). Simultaneous exposure to beta-araA and hypertonic medium increased the yield of interphase chromosome breaks further to 5.3 breaks/cell/Gy. This increase was consistent with an additive effect of each treatment on the overall yield of breaks, and suggested that hypertonic medium and beta-araA affect distinct and independent subsets of radiation-induced interphase chromosome breaks. We tested further the notion of independence by measuring rejoining of interphase chromosome breaks sensitive to hypertonic treatment in the presence of 1 mM beta-araA, and vice versa, rejoining of interphase chromosome breaks sensitive to beta-araA during and after treatment in hypertonic medium (500 mM NaCl, 20 min); under both sets of conditions each treatment caused maximal expression of prematurely condensed chromosome breaks responding sensitively to it when given immediately after irradiation. There was no change in the rejoining kinetics of interphase chromosome breaks sensitive to hypertonic treatment in the presence of beta-araA, and no change in the rejoining kinetics of interphase chromosome breaks sensitive to beta-arA in cells treated in hypertonic medium. These results are consistent with the hypothesis that exposure to ionizing radiation leads to the induction of two forms of prematurely condensed chromosome breaks that can be distinguished from each other on the basis of their repair kinetics and their differential sensitivity to treatment with beta-araA or hypertonic medium. In direct analogy to a classification proposed previously for potentially lethal damage (PLD) based on a similar set of experiments, we introduce the terms alpha form and beta form of interphase chromosome breaks for the slow, beta-araA-sensitive, and the fast, hypertonic treatment-sensitive form, respectively. We also propose that there is a correlation between alpha and beta form of interphase chromosome breaks and alpha and beta form of PLD, and present evidence suggesting that fast and slowly repairing DNA double-strand breaks underlie fast and slowly repairing interphase chromosome breaks.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8248485 TI - The effects of hypoxia and cysteamine on X-ray mutagenesis in human cells. II. hprt mRNA expression and cDNA sequence analysis of induced mutants. AB - Mutants at the hprt locus isolated after treatment of cells of the human lymphoblastoid cell line TK6 with X rays were examined by Northern blot and cDNA sequence analysis. In previous work, Southern blot analysis showed that approximately 25% of the mutants isolated from cultures treated with X rays displayed restriction fragment patterns indistinguishable from wild type. In addition, 38 and 48% of the mutants isolated from cultures treated with X rays under two radioprotective conditions, hypoxia or 25 mM cysteamine, respectively, had normal restriction fragment patterns. In the work presented here, Northern blot and DNA sequence analyses were used to characterize these mutants further. Mutants were classified as having normal size and amount of hprt mRNA, reduced amount or undetectable mRNA, or abnormal size message. Mutants that expressed hprt mRNA were sequenced after reverse transcription and PCR amplification. Sequence analysis is reported for 7 mutants from cultures treated in the absence of protection, 11 from cultures treated under hypoxic conditions, and 11 from cultures irradiated in cysteamine. Striking differences among the three treatment groups were not apparent. All types of mutations at both AT and GC base pairs were observed, including transitions, transversions, small deletions, and insertions. Several mutations affected RNA splicing, leading to exon skipping or inclusion of intron sequences in the final message. Approximately half (14/29) of the mutants sequenced had additions or deletions of one to several nucleotides. Also, 3/29 involved tandem DNA base changes (GG-->TT, C-->AA, AA-->CG). These observations are consistent with a mechanism involving the induction of noncoding or synthesis-blocking lesions that result in polymerase slippage or error-prone bypass synthesis. In addition, potential hotspot sites for mutation by X rays were discovered. At one site in exon 3, the same complex mutation, consisting of a G-->T transversion and a nearby six-base deletion, was detected in three independent mutants. Another mutant had a G-->C transversion at the same base, but without the deletion. At another site in exon 8, three mutations occurred at a run of three consecutive cytosines; these included a -C, a -CC, and a C-->AA. Also in exon 8, two mutations (+T, T-->C) occurred at two consecutive thymines. PMID- 8248486 TI - Histological changes in mouse colon after single- and split-dose irradiation. AB - These studies were undertaken to determine the relationship between acute mucosal damage and late obstructions in the colorectal region in the mouse after exposure to radiation. Radiation doses that either permanently depleted the mucosal epithelial cells or spared the mucosal lining by allowing epithelial regeneration were used. The distal 2.5 cm of colon and rectum of male C3Hf/Kam mice was irradiated with either a range of single doses (15-35 Gy) or two equal doses ranging from 9.75 to 14.75 Gy separated by 10 days. The time of onset and the incidence of obstructions and strictures in the bowel were recorded as a function of dose and time after irradiation. Acute damage in the mucosa and subsequent histological changes in the bowel were documented by sequential histological studies. Doses greater than 20 Gy caused acute crypt depletion followed by nonproductive attempts at regeneration and repopulation that culminated in persistent epithelial denudation. In these mice, obstructions appeared as early as 4 weeks and were characterized histologically by a mucosal ulceration extending deep into the muscularis. Single doses of less than 20 Gy and the split doses produced acute crypt cell depletion followed by successful regeneration, repopulation, and restoration of the colonic mucosa. In these mice, obstructions did not appear until at least 40 weeks after irradiation and were characterized by an intact mucosa with a thickened and fibrotic submucosa. Animals given a single dose of 20 Gy developed obstructions throughout the duration of the experiment. Those obstructions that occurred before 6 months were characterized by ulcerations, whereas those that appeared after this time exhibited only fibrosis in the submucosa with no mucosal ulceration. Based on these data, we suggest that two types of late obstructions occur in the bowel, one that depends on persistent epithelial denudation, i.e., a "consequential" response, and the other in the absence of epithelial denudation, i.e., a true late effect. PMID- 8248487 TI - Induction of hprt mutations in mice after exposure to fission-spectrum neutrons or 60Co gamma rays. AB - The effects of exposure to fission-spectrum neutrons and 60Co gamma rays on mutation induction in B6CF1 mice were investigated. Mutation induction was measured at the hypoxanthine-phosphoribosyl-transferase (hprt) locus in splenic lymphocytes at 56 days after whole-body irradiation. Lymphocytes were cultured 12 16 days in round-bottomed, 96-microwell plates in the presence of 5 x 10(4) feeder cells (syngeneic lymphocytes irradiated with 50 Gy gamma rays). The selective agent used as 6-thioguanine at a concentration of 2.5 micrograms/ml. Animals were exposed to either single doses of neutrons (1.5 Gy) or photons (7.5 Gy) or fractionated doses delivered over 2 weeks of neutrons (0.25 Gy x 6, total 1.5 Gy) or photons (1.5 Gy x 6, total 9.0 Gy). The frequency of hprt mutant induction by fission-spectrum neutrons delivered in a 1.5-Gy single dose compared to a 7.5-Gy single dose of 60Co photons was approximately the same, i.e., 5.98 x 10(-5) +/- 1.51 x 10(-5) (SE) vs. 5.56 x 10(-5) +/- 3.09 x 10(-5) (SE), respectively (Student's two-tailed t test, P = 0.8997). Multiple doses of neutrons gave rise to slightly higher mutant frequencies compared to photons even though the ratio of total doses of neutrons to photons was increased from 5 to 6, i.e., 8.71 x 10(-5) +/- 5.39 x 10(-5) (SE), total dose 1.5 Gy, vs 2.30 x 10(-5) +/- 9.07 x 10(-6) (SE), total dose 9.0 Gy, respectively (Student's two-tailed t test, P = 0.3330). These results suggest that the relative differences in magnitude in radiation-induced genotoxic effects between fission-spectrum neutrons and 60Co gamma rays increase when the comparisons are made between fractionated rather than single-dose exposure regimens. PMID- 8248488 TI - Is there an adaptive response to radiation in the developing brain of the fetal rat? AB - An adaptive response has been demonstrated in certain mammalian cells wherein pre exposure to a small radiation dose prior to a large dose ameliorates the damage induced by the second dose. We investigated whether a similar response could occur in the developing brain of the fetal rat, and if so, what the optimum interval between the two doses would be. Pregnant rats were exposed to a dose of 0.02 Gy gamma radiation at variable times (1, 3, 6, 12, or 24 h) prior to a second dose of 0.5 Gy on day 15 of gestation. Fetuses were harvested at 6 and 24 h after the second irradiation and standard cellular morphological assessments performed on the developing cerebral cortex. For number of mitotic cells, pyknotic cells, and macrophages, no significant differences were found between any of the groups that had received the priming (0.02 Gy) dose and the group that had not. Significant differences were found between fetuses harvested at 6 h and those harvested at 24 h after the final irradiation for all parameters measured. Thus, while the data were consistent with past research relating to the effects of radiation on the development of the brain of the fetal rat, no evidence for an adaptive response to radiation was found. Whether an adaptive response was indeed absent, or whether the doses and/or intervals used were simply not appropriate for demonstrating it, remains unknown. PMID- 8248489 TI - Cell survival from single passages for high-LET track-segment experiments with cell monolayers. PMID- 8248490 TI - Points on comparisons of cellular radiation response. PMID- 8248491 TI - Leukemia risk in thorotrast patients. PMID- 8248492 TI - Can nurses testify as to physician malpractice? PMID- 8248493 TI - Nurses fail to listen to patient's complaints. Case in point: Harmon v. Patel 617 N.E. 2d 183--IL (1993). PMID- 8248494 TI - Legal case briefs for nurses. MO: home "controls" private nurse's aide: workers' compensation liability results; MA: drunken patient admitted to E.R.: failure to monitor patient--death results. PMID- 8248495 TI - Failure to repeat vitals--paralysis: proximate cause issue. Case in point: Porter v. Lima Memorial Hospital 995 F. 2d 629--OH (1993). PMID- 8248496 TI - Cholinergic neurons identified by in situ hybridization histochemistry. PMID- 8248497 TI - Acetylcholine receptors: drugs and molecular genetics. PMID- 8248498 TI - Functional diversity of neuronal nicotinic acetylcholine receptors. PMID- 8248499 TI - Muscarinic acetylcholine receptor subtypes: localization and structure/function. AB - Based on the sequence of the five cloned muscarinic receptor subtypes (m1-m5), subtype selective antibody and cDNA probes have been prepared. Use of these probes has demonstrated that each of the five subtypes has a markedly distinct distribution within the brain and among peripheral tissues. The distributions of these subtypes and their potential physiological roles are discussed. By use of molecular genetic manipulation of cloned muscarinic receptor cDNAs, the regions of muscarinic receptors that specify G-protein coupling and ligand binding have been defined in several recent studies. Overall, these studies have shown that amino acids within the third cytoplasmic loop of the receptors define their selectivities for different G-proteins and that multiple discontinuous epitopes contribute to their selectivities for different ligands. The residues that contribute to ligand binding and G-protein coupling are described, as well as the implied structures of these functional domains. PMID- 8248500 TI - Molecular biology of cholinesterases: a background and an introduction. PMID- 8248501 TI - Structure and functions of acetylcholinesterase and butyrylcholinesterase. PMID- 8248502 TI - Promoter elements and alternative splicing in the human ACHE gene. PMID- 8248503 TI - The cholinesterases: a discussion of some unanswered questions. PMID- 8248504 TI - Regulation of the synthesis of acetylcholine. PMID- 8248505 TI - Molecular genetic specification of cholinergic neurons. PMID- 8248506 TI - Acetylcholine transporter--vesamicol receptor pharmacology and structure. PMID- 8248507 TI - Storage and release of acetylcholine in a sympathetic ganglion. PMID- 8248508 TI - Acetylcholine turnover and release: the influence of energy metabolism and systemic choline availability. PMID- 8248509 TI - Choline, a precursor of acetylcholine and phospholipids in the brain. PMID- 8248510 TI - The non-quantal release of acetylcholine from motor nerve terminals: comment on its likely size. PMID- 8248511 TI - Molecular aspects of acetylcholine release: an overview. PMID- 8248512 TI - Molecular approaches to synaptic vesicle exocytosis. PMID- 8248513 TI - The neurotrophic hypothesis and the cholinergic basal forebrain projection. PMID- 8248514 TI - Nerve growth factor affects the cholinergic neurochemistry and behavior of aged rats. PMID- 8248515 TI - Protective effects of nerve growth factor and brain-derived neurotrophic factor on basal forebrain cholinergic neurons in adult rats with partial fimbrial transections. PMID- 8248516 TI - Trophic responses of forebrain cholinergic neurons: a discussion. PMID- 8248517 TI - Comments to session on electrophysiological aspects of cholinergic mechanisms. PMID- 8248518 TI - Central cholinergic mechanisms and function. PMID- 8248519 TI - Actions of acetylcholine in the cerebral cortex and thalamus and implications for function. PMID- 8248520 TI - The phosphoinositide signalling system. I. Historical background. II. Effects of lithium on the accumulation of second messenger inositol 1,4,5-trisphosphate in brain cortex slices. PMID- 8248521 TI - Lithium selectively potentiates cholinergic activity in rat brain. PMID- 8248522 TI - Characteristics of the changes in intracellular calcium concentration on the activation of muscarinic receptors in hippocampal neurons. PMID- 8248523 TI - Cholinergic blockage of network- and intrinsically generated slow oscillations promotes waking and REM sleep activity patterns in thalamic and cortical neurons. PMID- 8248524 TI - Cholinergic modulation of sensory information. PMID- 8248525 TI - The cholinergic neuromodulatory system: an evaluation of its functional roles. PMID- 8248526 TI - Cholinergic receptor subtypes and REM sleep in animals and normal controls. AB - As reviewed here and elsewhere in this symposium, acetylcholine, in conjunction with other neurotransmitter systems, plays a very important role in the regulation of circadian and sleep-wake states. To briefly recapitulate, several current basic concepts about the regulation of sleep-wake states include: (a) REM sleep, or at least its phasic events (eye movements and PGO spikes), are promoted by cholinergic neurons originating within the peribrachial regions [LDT/PPT] (Mitani et al., 1988; Shiromani et al., 1988; Datta et al., 1991; Shouse and Siegel, 1992); (b) REM sleep may be inhibited by noradrenergic and serotonergic neurons in the locus coeruleus and dorsal raphe, respectively (Siegel, 1989; Steriade and McCarley, 1990; Jones, 1991); (c) stages 3 and 4 (Delta) sleep are inhibited by cholinergic terminals from basal forebrain to cortex (Buzsaki et al., 1988) and from LDT/PPT to thalamus (Steriade and McCarley, 1990; Steriade et al., 1991); (d) Delta sleep is modulated by complex serotonergic mechanisms; for example, it is increased by pharmacological antagonists of 5HT2 receptors (Declerck et al., 1987; Dugovic et al., 1989; Benson et al., 1991), although the mechanism and neuroanatomical site at which this effect occurs is unknown. Given the importance of mACHR mediation of components of REM sleep, it is unfortunate that so little is known about the distribution of the various subtypes of mACHRs in brainstem areas which regulate REM sleep. mACHR subtypes have been identified by molecular, biological and pharmacological methods.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248527 TI - Acetylcholine as a brain state modulator: triggering and long-term regulation of REM sleep. PMID- 8248528 TI - Behavioral aspects of cholinergic transmission: role of basal forebrain cholinergic system in learning and memory. PMID- 8248529 TI - Role of forebrain cholinergic systems in learning and memory: relevance to the cognitive deficits of aging and Alzheimer's dementia. PMID- 8248530 TI - Clinical aspects of cholinergic pharmacology. PMID- 8248531 TI - Approaches to cholinergic therapy in Alzheimer's disease. PMID- 8248532 TI - What have we learned from the THA trials to facilitate testing of new AChE inhibitors. PMID- 8248533 TI - Brain selective inhibition of acetylcholinesterase: a novel approach to therapy for Alzheimer's disease. AB - It could be argued that clinical experience with cholinergic drugs in the therapy of AD has not yet shown relevant symptomatic improvements. The main reasons for this might be attributed to peripheral cholinergic effects and the liver toxicity of some of these drugs, which limit their use and prevent confirmation of the cholinergic hypothesis (Gray et al., 1989). The main disadvantages of the cholinesterase inhibitors used in clinical trials are the short duration of action in the case of physostigmine and the potential for liver toxicity seen with the aminoacridine derivatives. The results presented with SDZ ENA 713 indicate that the disadvantages of AChE inhibitors might be overcome by improving CNS selectivity and thereby decreasing the peripheral cholinergic effects and toxicity. Clinico-pharmacological studies with SDZ ENA 713 have been performed in healthy volunteers; while central activity was clearly demonstrated in an EEG sleep study (Holsboer et al., 1992), no prohibitive peripheral side effects were seen, confirming in humans the results obtained in experimental animals (Enz et al., 1991). A multicentre clinical investigation in AD patients has been performed in Europe and is currently being evaluated. PMID- 8248534 TI - Subtype selective muscarinic agonists: potential therapeutic agents for Alzheimer's disease. PMID- 8248535 TI - Pharmacotherapy of Alzheimer disease: new drugs and novel strategies. PMID- 8248536 TI - Overview and future directions of CNS cholinergic mechanisms. PMID- 8248537 TI - Cholinergic systems: human diseases, animal models, and prospects for therapy. PMID- 8248538 TI - The organization of central cholinergic systems and their functional importance in sleep-waking states. AB - Since the demonstration some 50 years ago of the presence and synthesis of acetylcholine (ACh) in specific neuronal systems within the brain, a wealth of information concerning the organization and functional importance of central cholinergic neurons has emerged through immunohistochemical, neuroanatomical, pharmacological, biochemical and neurophysiological studies. Many of the original theses have proven valid concerning the key structural and functional position of cholinergic neurons within the central reticular core of the brain, where the basic sleep-waking cycle is determined. The two major cholinergic cell groups of this core, one within the pontomesencephalic tegmentum that projects rostrally into the non-specific thalamo-cortical relay system and the other within the basal forebrain that receives input from the brainstem reticular formation and projects in turn as the ventral, extrathalamic relay upon the cerebral cortex, are critically involved in processes of cerebral activation that accompany the states of wakefulness and paradoxical sleep. By interaction with other cell groups, including monoaminergic and GABAergic neurons, and by differential modes of firing, the cholinergic neurons may furthermore shape the responsiveness and activity of the reticular core and thalamo-cortical systems across the sleep waking cycle. PMID- 8248539 TI - CNS distribution of cholinergic receptors--some questions from a clinical neuroscientist. PMID- 8248540 TI - Nicotinic receptors in mammalian brain: localization and relation to cholinergic innervation. PMID- 8248541 TI - Autoradiographic distribution of putative muscarinic receptor sub-types in mammalian brain. PMID- 8248542 TI - Advances and limitations of the molecular neuroanatomy of cholinergic receptors: the example of multiple muscarinic receptors. PMID- 8248543 TI - Macrophage heterogeneity established by immunocytochemistry. PMID- 8248544 TI - Inhibition of antigen-induced airway hyperresponsiveness by a thromboxane A2 receptor antagonist (AA-2414) in Ascaris suum-allergic dogs. AB - We studied changes in airway responsiveness to acetylcholine (ACh) after antigen inhalation in Ascaris suum (A. suum)-allergic dogs. Airway responsiveness was determined by obtaining a dose-response curve of lung resistance plotted against increasing concentrations of ACh aerosol before and after inhalation of A. suum antigen. To determine the role of thromboxane A2 (TXA2) in the airway response, we tested the effect of a TXA2 receptor antagonist, AA-2414, in A. suum-allergic dogs. The procedure was repeated in each dog at an interval of 2 weeks to evaluate the effect of AA-2414 in a crossover manner. The dogs showing an airway response to antigen showed an increase in airway responsiveness to ACh 2, 4 and 6 h after antigen inhalation. The increase in airway responsiveness was significantly inhibited by administration of AA-2414 (5 mg/kg, i.v.) before antigen inhalation. These results suggest that TXA2 may be involved in antigen induced airway hyperresponsiveness (AHR) in dogs. PMID- 8248545 TI - Increased thromboxane A2 synthesis by rat lung neutrophils during selenium deficiency. AB - Modulation of cellular hydroperoxide levels is considered one of the important physiological mechanisms for regulating the synthesis of prostaglandins (PGs) and leukotrienes (LTs) in mammalian cells. Both vitamin E and selenium (Se) have the potential to affect the concentration of peroxides and, thus, the biosynthesis of eicosanoids. To gain insight into some of the molecular mechanisms underlying the regulation of the arachidonic acid cascade by vitamin E and Se, we have investigated the influence of altered vitamin E and Se nutrition on the ability of polymorphonuclear leukocytes (PMNs) derived from endotoxin-challenged lung to secrete arachidonic acid metabolites. Selenium deficiency had no significant effect (p > 0.05) on lavage fluid levels of thromboxane (TX) B2, LTB4 or LTC4. Vitamin E deficiency, however, led to a significant increase in LTB4 recovered from lavage fluid while having no effect on TXB2. In contrast, Se deficiency, although producing no discernible effects on the production of LTB4, resulted in a significant increase in the release of TXB2 by PMNs. An increase in TXB2 release was seen in both in vitro-stimulated and nonstimulated PMNs. Vitamin E deficiency appeared to induce an enhancement of LTB4 release by PMNs but the increase was not statistically significant. No detectable levels of LTC4 were found in PMN cultures stimulated with either zymosan or A23187. Thus, these studies indicate that deficiencies of either Se or vitamin E lead to alterations in the metabolism of arachidonic acid in the lung. PMID- 8248546 TI - De novo synthesis of phospholipase A2 and prostacyclin production by proliferating rat smooth muscle cells. AB - We investigated the role of phospholipase A2 (PLA2) in cell cycle-dependent alterations of endogenous prostacyclin (PGI2) synthesis in aortic smooth muscle cells in culture (VSMC) from Wistar Kyoto rats. Randomly cycling VSMC generated more PGI2 than the stationary cells. Cell cycle analysis showed that PGI2 production capacity was increased from the G0/G1 through the early DNA synthetic (S) phases. Enzyme analysis revealed that, although there were different mechanisms underlying this increase in the PGI2 production during the G0/G1, the peak at 4 hours coincided with a sharp increase in PLA2 activity. This increase in PLA2 activity was preceded by an increased expression of functional PLA2 messenger RNA, and protein synthesis inhibition prevented most of the increase in PGI2 production at 4 hours. These data indicate that endogenous PGI2 generation is mainly increased during the G0/G1 period and that this event is secondary to de novo synthesis of PLA2 and probably, at least in part, to cyclooxygenase induction. This mechanism provides a negative feedback regulating VSMC proliferation. PMID- 8248547 TI - A comparison of serum and vaginal dialysate concentration of 13,14-dihydro-15 keto-PGF2 alpha in women undergoing 1st trimester therapeutic abortions. AB - 13,14-Dihydro-15-keto-PGF2 alpha(PGFM) levels in serum and vaginal secretion, before, one hour, and 24 hours post abortion were measured in 13 women from 8 to 12 weeks pregnant. The mean metabolite levels in the blood were 61.2 pg/ml pre termination, 73.1 pg/ml and 121.4 pg/ml one hour and 24 hours post-termination. The vaginal dialysate means were 136.7 pg/ml, 425.7 pg/ml, and 211.7 pg/ml respectively. PMID- 8248548 TI - Effects of interleukin-6 and tumor necrosis factor-alpha on prostaglandin production by cultured human fetal membranes. AB - In this study we investigated the effects of the cytokines interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) on prostaglandin production by cultured human fetal membranes. These cytokines stimulate prostaglandin synthesis by isolated components of human fetal membranes, but their effects on the intact tissue comprising amnion, chorion and decidua were not known. TNF-alpha added to the maternal side of the membrane activated decidual production of PGF2 alpha but had no effects on synthesis of PGE2 or PGE2 metabolites. Addition of TNF-alpha to the fetal side of the membrane increased production of PGE2 by amnion and PGE2 metabolites from chorion. The addition of IL-6 to the fetal or the maternal side of the membrane increased production of PGE2 from amnion and PGE2m from chorion, suggesting that IL-6 might pass through the fetal membrane. IL-6 had no effect on decidual PGF2 alpha production. These results suggest that TNF-alpha may be involved in labor by increasing decidual prostaglandin synthesis, whereas IL-6 is less likely to have a role. PMID- 8248549 TI - Effect of IGF-1 on placental prostanoid production. AB - Prostanoids play an important role throughout all of pregnancy and during the initiation and progress of labor. The human placenta, at term, produces large quantities of prostanoids, yet little is known of the steps regulating their biosynthesis. In these studies, the effect of IGF-I on the release of placental prostanoids was investigated. The basal release of prostaglandin E (PGE), prostaglandin F (PGF), thromboxane (TxB2) and 6-keto-prostaglandin F1 alpha (6 keto-PGF1 alpha) increased from the fifth hour in culture, while the release of 13, 14-dihydro-15-keto-PGF2 alpha (PGFM) remained constant and hCG release decreased. The addition of IGF-I (10(-8) M) to the perfusing medium effected an inhibition of TxB2 and PGF within two and one-half hours of exposure. However, the release of PGE, PGFM, 6-keto-PGF1 alpha or hCG was not altered by IGF-I. Because both TxB2 and PGF are vaso-constrictors, we have proposed that IGF-I may enhance vasodilation in the placenta. Therefore, IGF-I may allow increased blood flow, thus affecting the maintenance of pregnancy and supply of nutrients for the growth of the fetus. PMID- 8248550 TI - Identification of 19 (R)-OH prostaglandin E2 as a selective prostanoid EP2 receptor agonist. AB - The physiological significance of the formation of large quantities of 19(R) hydroxy prostaglandin E's (19-OH PGE) from PGE1 and PGE2 in human seminal plasma is intriguing. The concept that prostaglandins exert their biological effects by interacting with specific receptors, according to the current working classification for prostanoid receptors, was employed as a conceptual framework to re-examine the activity of 19(R)-OH PG's. In contrast to PGE2, which may indiscriminately stimulate a variety of prostanoid receptor subtypes, 19(R)-OH PGE2 exhibited selectivity for the EP2-receptor subtype. In EP1 (guinea pig ileum contraction), EP2 (cat trachea relaxation), and EP3 (chick ileum contraction) preparations where PGE2 is equipotent, 19(R)-OH PGE2 exhibited greater potency in the EP2-receptor population. Moreover, unlike PGE2, 19(R)-OH PGE2 did not stimulate an FP-receptor preparation (cat iris). 19(R)-OH PGE2 was devoid of activity at thromboxane A2-(TP), prostaglandin D2-(DP) and prostacyclin-(IP) sensitive receptors as indicated by its inability to cause human platelet aggregation or inhibit ADP-induced platelet aggregation. 19(R)-OH PGE1 had an entirely converse profile of activity. As a myotropic agent in the guinea pig and chick ileal preparations, 19(R)-OH PGE1 was approximately 1.5 orders of magnitude more potent than 19(R)-OH PGE2 but it appeared devoid of EP2-receptor stimulant properties. 19(R)-OH PGF2 alpha possessed very little biological activity in a diverse variety of isolated tissue preparations, indicating that 19-hydroxylation represents a highly efficient inactivation step for PGF2 alpha. The implications of the formation of receptor selective PGE derivatives in human seminal fluid for human reproductive physiology remains to be established. PMID- 8248551 TI - A model for calculating the costs of in vivo dosimetry and portal imaging in radiotherapy departments. AB - The costs of in vivo dosimetry and portal imaging in radiotherapy are estimated, on the basis of a detailed overview of the activities involved in both quality assurance techniques. These activities require the availability of equipment, the use of material and workload. The cost calculations allow to conclude that for most departments in vivo dosimetry with diodes will be a cheaper alternative than in vivo dosimetry with TLD-meters. Whether TLD measurements can be performed cheaper with an automatic reader (with a higher equipment cost, but lower workload) or with a semi-automatic reader (lower equipment cost, but higher workload), depends on the number of checks in the department. LSP-systems (with a very high equipment cost) as well as on-line imaging systems will be cheaper portal imaging techniques than conventional port films (with high material costs) for large departments, or for smaller departments that perform frequent volume checks. PMID- 8248552 TI - HDR versus LDR gynecological brachytherapy revisited. AB - Despite the obvious breakthrough of high dose-rate (HDR) afterloading systems on the gynecological brachytherapy market, questions still remain regarding the transfer of available expertise gained throughout the last 80 years with low dose rate (LDR) radium and cesium, especially regarding the conversion of LDR total dose into equivalent HDR dose per fraction and total dose. Calculation of biologically equivalent schedules requires a knowledge of repair capacity and repair kinetics of tumors and normal tissues, both of which influence the biological effect of any radiation dose. The clinical experience with HDR is, however, accumulating and it is acknowledged that the new technique entails an acceptable therapeutic index as compared to the classical LDR. There is thus a state of apparent 'equivalence' between the two treatment modalities. This state is influenced by many factors in which, in contrast to what is frequently claimed, radiobiological factors do not play the most important role. It is probably its high-tech environment which makes HDR an acceptable alternative. Treatment at LDR, indeed, has proven to be quite tolerant to a lack of absolute precision, something that would be disastrous with HDR techniques. Because HDR intracavitary brachytherapy has not been compared in controlled trials with the best existing LDR brachytherapy, but only retrospectively with heterogeneous LDR clinical data, it cannot be claimed to be equivalent, but simply feasible. PMID- 8248553 TI - Intraoperative radiotherapy in recurrent gynecological cancer. AB - A retrospective analysis to assess the feasibility and clinical tolerance of intraoperative radiotherapy (IORT) in the treatment of recurrent gynecologic cancer is reported. From February 1985 to September 1992, 26 patients with recurrent gynecologic tumors entered this trial. The clinical experience comprises two different categories of disease situations: tumors relapsing after full dose radiation therapy (group I) and recurrent disease to previous surgery (group II). Cervical carcinoma was the initial tumor site of involvement in 18 patients (69%). Treatment consisted in maximal surgical resection + IORT boost (10-25 Gy) to the high-risk areas for recurrence. Non previously irradiated patients also received external beam irradiation (EBRT) (+/- chemotherapy) pre- or postoperatively. IORT-related toxicity was one episode of motor neuropathy. Local control rates have been 33% and 77%, respectively in groups I and II. The 4 year actuarial overall survival in Group I is 7% and 6-year actuarial overall survival in Group II is 33%. The addition of IORT to surgical debulking achieves modest local control and long-term survival rates if tumor-free margins cannot be obtained in previously irradiated patients. Combined EBRT (+/- chemotherapy) maximal surgical resection plus IORT could render some long-term survivors among those surgical recurrent patients not candidates for radical surgery with curative intent. PMID- 8248554 TI - Reporting radiotherapeutic complications in patients with uterine cervical cancer. The importance of latency and classification system. AB - Radiotherapeutic morbidity is reported according to our own system (AADK) and the Franco-Italian glossary (FI) in 442 patients with cervical cancer FIGO stage IIB (139), IIIA (10), IIIB (221), and IVA (72). The AADK system records each symptom, date of appearance, the required therapy, and its initial date. FI describes the maximal damage in 4 grades. Actuarial estimates of moderate or worse complications in the rectosigmoideum differed significantly in relation to stage, while frequencies did not differ. Frequencies were up to 25% lower than the actuarial estimates. Moderate AADK complications in the rectosigmoideum occurred from 1 to more than 24 months in 42% of stage IIIB patients finally developing severe FI complications, and during more than 2 years in 24% of the patients dying from rectosigmoid complications. An analysis of the probability of being alive without moderate or worse AADK complications indicated that survival and complications were unrelated. It is concluded that, with any classification system for reporting morbidity, each symptom and required therapy used in the definition of each complication grade and the date of appearance should be registered regularly to allow (1) reporting of the real risk of organ damage, (2) rescoring of complication grades, (3) separation of early and late morbidity, and (4) reporting of actuarial estimates. If these minimum requirements are met, underestimation of morbidity is avoided. PMID- 8248555 TI - Squamous cell carcinoma of the lip: analysis of the Princess Margaret Hospital experience. AB - We reviewed 117 patients with squamous cell carcinoma of the lip who were treated at the Princess Margaret Hospital between 1976 and 1985. Ninety-eight cancers arose from the lower lip, 18 from the upper lip and 1 from the commissure. Two patients had lymph node metastases at presentation. Sixty-one patients were treated with radiation therapy following a biopsy, 28 underwent surgery followed by post-operative radiation, and 28 had surgery alone. With a median follow-up time of 5.4 years, the 5-year actuarial overall and cause-specific survival rates were 81% and 99%, respectively. Local failure developed in 4 patients after radiation treatment, 3 of whom were salvaged by surgery. Six patients developed regional metastases after initial treatment, 4 of whom were salvaged with surgery and/or radiotherapy. Two patients died from lip cancer. After a univariate analysis, the only factor which predicted for nodal failure was T stage of the primary lesion, with a 4% risk of nodal failure for T1 lesions vs. 20% for T2/3 lesions (p = 0.03). No other patient, tumour or treatment variables influenced loco-regional control or survival in a statistically significant manner. Cosmetic and functional outcome were evaluated in 8 patients whose radiation treatments were administered 13 years ago. No patients had compromised lip function, and the majority had minimal cosmetic sequelae from their radiation therapy. Based on the excellent results of this review, we would continue to recommend radiation therapy as an effective treatment modality for patients with lip cancer because of the ease by which the entire tumour can be encompassed whilst maintaining excellent cosmetic and functional outcome. PMID- 8248556 TI - Rates of local control, metastasis, and overall survival in patients with posterior uveal melanomas treated with ruthenium-106 plaques. AB - The prognostic importance of clinically assessed tumor variables, patient variables, and treatment variables of 184 patients treated with ruthenium-106 eye plaques for uveal melanoma was studied using crude survival, metastatic disease and local tumor control as the endpoints. Using the multivariate Cox proportional hazards model, tumor diameter, age and sex were identified as the strongest variables predicting survival. Risk of metastasis was found to be dependent on tumor size (diameter) only and the metastasis was generally observed within 6 years after the end of treatment (96%). Clinically diagnosed local tumor control was predicted by tumor T-stage, treatment time, and total dose to the sclera. No correlation between metastatic disease and local failure was detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248557 TI - Hypofractionation in retinoblastoma: an increased risk of retinopathy. AB - Forty-four eyes in 38 children were treated between 1963 and 1991 by external radiotherapy for retinoblastoma. Treatment modalities varied widely during this period; in addition to radiotherapy there was chemotherapy (16/44), photocoagulation (14/44), and laser therapy or cryotherapy (14/44). Treatment technique and dose fractionation also varied widely; lateral beam technique (39/44) versus anterior or anterior/lateral beam; doses per fraction ranged from 1 to 4.5 Gy, total doses from 30 to 61.5 Gy, and overall times from 22 to 49 days. Patients were followed at 3-month intervals, and actuarial survival at 10 years was 88%, with 62% local control. Ten eyes showed clinical evidence of retinopathy. A multivariate analysis of factors associated with increased risk of retinopathy was carried out using the Cox proportional hazards model and the mixture model of Farewell. The estimated latent time was 17 months (95% confidence interval, 14-20 months). The only factors found to be significantly associated with retinopathy were total dose multiplied by dose per fraction, or total dose normalized to the equivalent total dose in 2-Gy fractions as estimated from the LQ model, and these gave equivalent descriptions. There were trends (not significant) for increased risk of retinopathy when treatments included chemotherapy or photocoagulation, and for decreased risk (also not significant) when cryotherapy was used in conjunction with radiotherapy. No significance could be attached to any of the following: number of sites per eye, Reese-Ellsworth stage, and family history. We conclude that hypofractionation carries a significant risk for retinopathy in the treatment of retinoblastoma. PMID- 8248558 TI - Radiosensitization of SCCVII tumours and normal tissues by nicotinamide and carbogen: analysis by micronucleus assay. AB - The radiosensitizing effect on SCCVII tumours of carbogen (95% O2 + 5% CO2) combined with nicotinamide was investigated using the micronucleus assay and an in vivo/in vitro colony formation assay following single irradiation. The effects on intestinal crypt cells and bone marrow cells were also examined in mice. The frequency of micronuclei in tumours increased with an increase in the nicotinamide dose (administered 1 h before the irradiation of 5 Gy) from 0.1 to 1.0 mg/g, and showed a 1.3-fold increase at 1.0 mg/g when compared with radiation alone. The micronucleus frequency showed a more marked increase following irradiation combined with nicotinamide and carbogen inhalation starting 15 min before irradiation. The radiosensitizing effect reached a plateau at a nicotinamide dose of 0.1 mg/g in combination with carbogen, giving an enhancement ratio (ER) of 1.8 relative to radiation alone at 2 Gy. In the radiation dose range of 5-20 Gy, ERs of 1.8-1.9 and 1.5 (at a 10% cell-survival level) were obtained for the combination of 0.1 or 1.0 mg/g nicotinamide and carbogen and for carbogen alone, respectively. A slight increase in the regeneration response of the jejunum and the bone marrow was observed following irradiation combined with 0.1 mg/g nicotinamide and carbogen, yielding ERs of 1.07 for the jejunum and 1.15 for the bone marrow. Thus, the nicotinamide/carbogen combination, with its large therapeutic gain factor at low doses and proven low toxicity in humans, seems to improve the response of cancer to radiotherapy. PMID- 8248559 TI - Comparison of treatment techniques for lung cancer. AB - A comparison of the treatment techniques for a typical case of lung cancer has been performed between 10 Swiss radiation oncology centers. The description and basic imaging data of a locally advanced non-metastatic lung cancer were sent to each institution and participants were asked to perform the treatment planning as for a routine case. The comparison shows that, in addition to a broad spectrum of treatment techniques, there was a significant variation in the major treatment parameters: definition of the target volume, prescription procedure, total dose to the tumor and dose per fraction. Although some reasons could be found to partially explain these differences, the lessons learned from this comparison are, first, that a standardization, in particular for dose prescription, is urgently needed, and, second, that a search for a better consensus in standard managements should be promoted in radiation oncology. PMID- 8248560 TI - ICRU reference points and maximum doses of rectum and bladder in intracavitary radiotherapy. AB - The absorbed doses in the reference points of the rectum and bladder defined by ICRU report 38 for reporting intracavitary radiotherapy were compared with the calculated maximum doses of these organs. The calculations were validated by a thermoluminescent probe. A good correlation between the doses at the reference points and the calculated maximum doses was found. PMID- 8248561 TI - Accidental overirradiation syndrome. AB - Between the 10th and 20th December 1990, there was a malfunction in the operation of the Electron Linear Accelerator (ELA) at the Hospital Clinico Universitario in Zaragoza (Spain), which led to irradiation with the highest beam energy. This situation shows the existence of a latent period between the irradiation and the appearance of the symptoms, even in situations involving much higher than normal doses in cases of localized irradiation. PMID- 8248562 TI - The effect of treatment duration in the local control of cervix cancer. PMID- 8248563 TI - Craniospinal radiotherapy: where does the thecal sac end? PMID- 8248564 TI - Regaud Lecture, Malmo, 1992. The clinical science of radiation oncology. AB - Claudius Regaud can be regarded as the founding father of both radiobiology and radiotherapy. His research extended widely in oncology and the conclusions which he drew from his work are considered in the light of our modern practice of the clinical science of radiation therapy. PMID- 8248565 TI - Possibilities and pitfalls in triangulation. PMID- 8248566 TI - The stress process in family caregivers of persons receiving chemotherapy. AB - The purpose of this analysis was to explore strain and depression among family caregivers of persons receiving chemotherapy for cancer using the Pearlin Stress Process Model. Seventy-five family caregivers of persons receiving chemotherapy participated, completing the Caregiver Strain Index and the Depression Subscale of the Profile of Mood States within 3 weeks of the initiation of chemotherapy. Hierarchical regression analyses indicated that 44% of the variance in strain and 40% of the variance in depression were explained by the antecedent variables of caregiver age and gender, patient age and gender, patient functional status, the presence of recurrent disease, perceived efficacy of coping strategies, and perceived adequacy of social support. Further analysis aimed at delineating the mechanisms through which coping and social support operate in the model found no evidence for a moderating or stress-buffering effect. However, coping mediated the relationship between strain and depression and social support mediated the relationship between functional status and depression. PMID- 8248567 TI - Social support sources and psychological well-being in older women with heart disease. AB - A test of the compensatory hierarchical model of social support sources was conducted in a sample of 80 older women with heart disease. Relationships of support sources to measures of psychological well-being also were investigated. Emotional and tangible support sources fit the model with the exception that unmarried women without children named friends more often than extended family as emotional support sources. Receipt of emotional support from nonfamily members rather than from sources including family members was related to lower positive affect. Similarly, receipt of tangible support from nonfamily members rather than from sources including family members was related to less satisfaction with life. Factors that disrupt the family relationships of older women with illness are worthy of further exploration. PMID- 8248568 TI - The effect of environmental sound and communication on CCU patients' heart rate and blood pressure. AB - The effects of high ambient stressors (equipment sounds) and social stressors (conversation) on heart rate (HR) and blood pressure (BP) were examined in coronary care patients. Simultaneously occurring sound level in decibels, actual sound, electrocardiogram, and BP were recorded three times/day over 2 days for 20 subjects. Repeated measures ANOVA and pairwise comparisons revealed maximum HR to be significantly higher during conversation than during low ambient sounds (quiet). High ambient stressors did not affect HR for these subjects. BP did not significantly change during any of the sound conditions. Further research is needed to delineate cardiovascular effects of specific social stressors. PMID- 8248569 TI - Cycle-phase changes in perceived stress in women with varying levels of premenstrual symptomatology. AB - Stress has been documented as a critical factor in understanding a variety of health problems. This study was designed to examine the relationships between symptom severity and perceived stress in women with varying severity levels of premenstrual symptoms. Perceived stress was evaluated before and after menstruation in 104 women. Women who had high levels of premenstrual symptomatology demonstrated significantly greater perceived stress in the premenstrual phase compared to the postmenstrual phase, while women with low levels of premenstrual symptomatology showed no cycle-phase difference. PMID- 8248570 TI - Mothers' self-reports of parenthood across the first 6 months postpartum. AB - A postpartum measure, What Being the Parent of a New Baby is Like, was administered to explore the development of maternal role for mothers (N = 76) at 1, 3, 4 1/2, and 6 months postpartum. Individual differences were relatively stable across time and accounted for much more variance than time effects. Mean scores on the Centrality (of baby) and Help (with parenting tasks) subscales decreased over time, and Evaluation (satisfaction with role performance) and Lifechange subscale scores increased. Primiparas demonstrated the steepest mean increase in Evaluation scores over time. Mean Evaluation scores for first- (n = 29) and second-time mothers (n = 33) were similar, but mothers of three or more children (n = 14) had significantly higher scores. Parity was also related inversely to amount of life change, and maternal education was related inversely to Evaluation and Centrality scores. Demographic, subscale, and contextual variable interrelationships were generally consistent with maternal role adaptation theory. Findings also suggest that parity effects are more complex than a dichotomous primipara/multipara representation. PMID- 8248571 TI - Attitudes of practicing nurses as predictors of intended care behavior with persons who are HIV positive: testing the Ajzen-Fishbein Theory of Reasoned Action. AB - The purpose of this descriptive correlational study was to examine practicing nurses' attitudes, subjective norms, and intentions to care for HIV positive patients, using the Theory of Reasoned Action. One hundred and forty-one subjects completed a questionnaire developed according to guidelines described by Ajzen and Fishbein (1980). Consistent with the theory, nurses' attitudes and subjective norms were found to be significant predictors of intentions to care for persons who are HIV positive (R2 = 0.27). Personal beliefs found to discriminate between intenders and nonintenders were those related to possible consequences for self, family, and friends, but not job-related consequences. Normative beliefs which discriminated between groups were also related to nonprofessional referents' expectations. In addition, qualitative data showed persistent concerns about occupational risk for contracting AIDS. Based on the results of this research, it is recommended that nurse educators in both clinical and academic settings, target specific educational/training interventions to include transmission, prevention, as well as exploration of feelings, attitudes, beliefs, and behavioral intentions about HIV-related topics. Further theory-based research and testing of interventions to change practicing nurses' attitudes and beliefs about HIV disease are advocated. PMID- 8248572 TI - The moderating effects of dispositional affectivity on relationships between job characteristics and nurses' job satisfaction. AB - Using the job satisfaction model embedded in the Price-Mueller nurse turnover model, the moderating effects of positive affectivity (PA) and negative affectivity (NA) on the relationships between the seven independent variables in the model and job satisfaction were examined. Multiple regression procedure was used to analyze data collected from 162 nursing employees of a 327-bed Veterans Administration Medical Center. The major determinants of job satisfaction were found to be routinization, instrumental communication, and PA. The impact of participation, instrumental communication, integration, distributive justice on job satisfaction were found to be moderated by PA. NA moderated the impact of promotional opportunity on job satisfaction. These predictors, in conjunction with demographic variables, explained 45% of the variance of job satisfaction. PMID- 8248573 TI - Single-item indicators in nursing research. AB - Single-item indicators that ask respondents for their global rating of a specific concept are congruent with nursing's emphasis on wholism and individualism. They allow the subject to take personally salient features of the situation into account when providing a response. The psychometric performance of single-item indicators in published research and in a sample data set using measures of the mother's choice and satisfaction with her employment decision support the validity and reliability of the measures, suggesting that these indicators deserve more attention in nursing research. Recommendations for the use of single item indicators are provided. PMID- 8248574 TI - [Radiologic research in Italy]. PMID- 8248575 TI - [Pulmonary tuberculosis today. Radiologic problems]. PMID- 8248576 TI - [Radiology of normal chest structures in the elderly patient]. PMID- 8248577 TI - [Vocal recognition in dental and oral radiology]. AB - Speech reporting benefits by units which can recognize sentences in any natural language in real time. The use of this method in the everyday practice of radiology departments shows its possible application fields. We used the speech recognition method to report orthopantomographic exams in order to evaluate the advantages the method offers to the management and quality of reporting the exams which are difficult to fit in other closed computed reporting systems. Both speech recognition and the conventional reporting method (tape recording and typewriting) were used to report 760 orthopantomographs. The average time needed to make the report, the legibility (or Flesch) index, as adapted for the Italian language, and finally a clinical index (the subjective opinion of 4 odontostomatologists) were evaluated for each exam, with both techniques. Moreover, errors in speech reporting (crude, human and overall errors) were also evaluated. The advantages of speech reporting consisted in the shorter time needed for the report to become available (2.24 vs 2.99 minutes) (p < 0.0005), in the improved Flesch index (30.62 vs 28.9) and in the clinical index. The data obtained from speech reporting in odontostomatologic radiology were useful not only to reduce the mean reporting time of orthopantomographic exams but also to improve report quality by reducing both grammar and transmission mistakes. However, the basic condition for such results to be obtained is the speaker's skills to make a good report. PMID- 8248578 TI - [Inverted axis scans in the study of the lumbo-sacral passage with computed tomography]. AB - In the CT study of the lumbosacral spine, the disk spaces must be scanned along a plane which parallels the relative intervertebral disk perfectly. However, the lumbosacral space often happens to be too oblique--i.e., over the possible inclination of the gantry (usually: +/- 25 degrees). Thus, the somatic planes of the spine overlap. Therefore, in a series of 1,800 patients, the lumbosacral space was studied with the gantry inclined in the opposite direction to that in conventional scans, with the maximum angle allowed by our unit (-25 degrees). The incidence of negative findings for disk conditions was 7% (versus 13.5% with conventional CT). In both the negative and the positive cases, inversion CT scans were diagnostic in 594 patients (468 with disk herniation)--i.e., in 33% of cases. PMID- 8248579 TI - [Achilles tendon ossification and achillodynia]. AB - Achilles tendon ossification is a rare and asymptomatic condition. The possible sequelae of tendinitis, bursitis and fracture are some of the causes of achillodynia. Achillodynia is often unrelieved by conservative management and excision of the ossification may be necessary. Moreover, ossification were found in 3 of 41 patients with spontaneous rupture of the Achilles tendon. Predisposing degenerative changes of the tendon were found in all patients. Thus there is no evidence that ossification by itself may increase the risk of tendon rupture. PMID- 8248580 TI - [Seat belt syndrome]. AB - The increasingly common use of seat belts in the standard features of cars has markedly reduced the seriousness of lesions due to motor vehicle accidents but, at the same time, has caused the appearance of a specific pattern of injuries strictly related to the type of restraint. Thus, the seat belt syndrome consists of soft-tissues, skeletal and visceral injuries associated with the use of the passenger restraint system. After describing the site of injuries, we report the most common features of traumatic lesions, also comparing the consequences of the three possible occurrences: unrestrained, with two- and three- point restraints. With lap seat belts, which are the less complex, injuries most frequently involve the abdomen and vertebrae while with the three-point variety of seat belts the chest and shoulder girdle are involved. No matter what the type of restraint, skin abrasions are nearly always observed, and associated with internal injuries in 30% of cases. From the analysis of distribution, type and seriousness of injuries we can summarize that: a) the use of passenger restraint systems has surely reduced mortality and morbidity associated with motor vehicle accidents; b) the seat belt will never be able to avoid traumatic events because strong forces act and strike in a very short time on small areas of textile fabric; c) seat belts can be still considered the easiest, most reliable and cost-effective preventive method. PMID- 8248581 TI - [Computed tomography in the study of periodontal disease]. AB - Periodontal disease, an affection of bacterial etiology, causes the destruction of the periodontal tissue. From both the diagnostic and the prognostic points of view, it is extremely important to evaluate the changes occurring in the alveolar bone in patients affected with periodontal disease. The information obtained by conventional radiographic techniques is usually poor due to the overlapping of anatomical structures and to the lack of an axial view of the structures. CT, thanks to millimetrical sections of the dental structures along the planes which parallel the hard palate, allows the evaluation of: 1) the size and the relationship between teeth roots, with no overlapping of anatomical structures; 2) bone thickness; 3) very early bone lesions around the teeth roots. The authors suggest CT as a valuable tool in the study of periodontal disease in its early stage and in the evaluation of advanced lesions, both before treatment and during follow-up, when it is important to check the results of conservative treatment. PMID- 8248582 TI - [The temporo-mandibular joint in pathologic conditions: rheumatoid arthritis and seronegative spondyloarthritis]. AB - Pain and dysfunction of the temporomandibular joint (TMJ) are major clinical problems, especially in arthritides and allied conditions. In the last 10 years, such new imaging methods as arthrography, CT and MRI have been developed, but many problems are still to be solved. This study was aimed at reporting the radiographic patterns of lesions in TMJ rheumatoid arthritis and seronegative spondyloarthropathies and at investigating the role of conventional radiology in the assessment of these disorders. Digital hypocycloidal tomography of the TMJ was performed on 44 patients affected with rheumatoid arthritis, on 2 with Sjogren's syndrome, 2 with ankylosing spondylitis, 4 with psoriatic arthritis and 1 with Reiter's syndrome. CT, with sagittal and coronal scans, was performed only on 9 patients (18 joints) whose clinical and radiographic findings were particularly severe. The radiographic features of the lesions--i.e., erosions, osteophytes, subchondral bone sclerosis and condylar-glenoid fossa remodelling- cannot be distinguished from one another and from the so-called allied conditions; nevertheless, in rheumatoid arthritis abnormalities are usually bilateral and symmetric, whereas in seronegative arthropathies joint involvement is usually unilateral. In the first stage of the disease, these lesions are better demonstrated by CT than by tomography, thanks to better contrast and spatial resolution of the former, especially on the lateral and medial aspects of the joint. The most common findings were: erosions (68.2%), osteophytes (31.8%), subchondral bone sclerosis (28.6%) and condylar-glenoid fossa remodelling (9.1%). In the clinical practice sagittal multidirectional tomography remains today the method of choice because it can demonstrate several areas on the condylar surface, with finer anatomical detailing, and it can solve most diagnostic problems in TMJ disorders. Thus, CT is recommended only in the patients whose diagnosis remains questionable with the above techniques. PMID- 8248583 TI - [Reference standards in the performance of a mammographic screening program. Results of the screening program in the province of Florence, 1992]. AB - The authors report the results of the Florence District program for the year 1992. 11,033 subjects were examined. Attendance rate (64.4%) was significantly related to age (42-49 = 71.0%; 50-59 = 66.7%; 60-70 = 58.5%). Recall rate to diagnostic assessment was 2.09% (mammographic abnormalities = 199, subjective symptoms other than pain = 32) and was related to age (42-49 = 3.12%; 50-59 = 1.65%; 60-70 = 1.77%). According to the results of diagnostic assessment 53 surgical biopsies (0.48%) were recommended and performed. The biopsy rate was also related to age (42-49 = 0.2%; 50-59 = 0.4%; 60-70 = 0.7%). Forty-seven carcinomas were detected in 46 subjects (benign/malignant biopsy ratio = 0.13). Cancer detection rate was 0.42% and changed significantly with age (42-49 = 0.13; 50-59 = 0.36; 60-70 = 0.68%), as well as the observed/expected cancer ratio (42 49 = 0.89; 50-59 = 2.03; 60-70 = 2.98). Detected cancers were nonpalpable in 57% of cases. Pathologic staging was pTIS in 2 cases, pT1a in 5, pT1b in 17, pT1c in 17, pT2 in 5, and pT4b in one case. Six of 47 (12.8%) cancers involved axillary nodes. A preliminary estimate of screening cost yielded a total cost of Lit. 397,671,000 for the year 1992--i.e., Lit. 36,000 per examined subject and Lit. 8,461,000 per detected cancer. The results are compared with reference standards for the evaluation of screening performance provided by the national breast screening program in the United Kingdom. PMID- 8248584 TI - [Computed tomography diagnosis of small intestine carcinoid]. AB - This study was aimed at evaluating CT diagnostic capabilities in detecting carcinoid tumors of the small bowel and comparing CT results with conventional radiographic findings. The CT diagnosis of carcinoid tumor was made in 6 cases based on CT findings. All patients had undergone conventional radiographs (transit or double-contrast studies of the small intestine) and 2 patients had undergone double-contrast barium enema of the colon. 500 ml of oral contrast medium were administered 2 hours, 1 hour and 15 minutes before CT scans, respectively. A hypotonic agent was injected i.v. immediately before the examination. Primary lesion extent, mesenteric involvement, extramesenteric lymph nodes and hepatic metastases were the investigated CT patterns. The first two variables were also assessed on plain radiographs. In all patients the diagnosis of enteric carcinoid tumor was confirmed at surgery or liver biopsy. CT proved to be useful in demonstrating the primary tumor in 5 cases, mesenteric involvement in all patients, liver metastases in 3, lymphadenopathy in none. CT yielded very accurate findings and thus allowed the diagnosis of carcinoid tumor to be made in all the 6 patients, while plain films resulted normal in 2 cases and aspecific in 4. In conclusion, CT, if adequately performed, proved the best technique to detect carcinoid tumors of the small bowel. Of course, further evidence is required. PMID- 8248585 TI - [Primary hepatocarcinoma. Diagnostic imaging and chemoembolization]. AB - The diagnostic and therapeutic approach by means of transcatheter arterial chemoembolization is analyzed in 100 patients affected with hepatocellular carcinoma (HCC): in all patients the HCC had been diagnosed by means of US-guided fine-needle biopsy: 75 of 100 patients were in Child A class, 24 in Child B and 1 in Child C class. The most sensitive imaging technique to identify HCC proved to be angiography (95%); post-Lipiodol CT was also of great value to stage the disease. The authors suggest a diagnostic-therapeutic approach including angiography and simultaneous chemoembolization right after US-guided biopsy, avoiding conventional CT; such a protocol allows high diagnostic accuracy at a lower cost. Chemoembolization proved to be a safe technique, with a low incidence of mortality and complications; the latter were easy to treat by medical therapy. The follow-up data relative to 39 of 100 patients who underwent 2 or more chemoembolization treatments prove the latter to be a valuable technique relative to both the evolution of HCC nodules (in 18/21 patients with single lesions and in 9/18 patients with multiple lesions, lesion size was unchanged or decreased) and to survival rates (70% in 65 patients followed-up 1 year at least); on the whole, 77 of 100 patients are still alive. PMID- 8248586 TI - [Arterial CT in the diagnosis of hepatocellular carcinoma: initial experience with 12 patients]. AB - Intra-arterial CT of the liver is a valuable method to evaluate hepatocellular carcinoma (HCC). It consists of an infusion of contrast medium into the hepatic artery during CT scanning. Twelve patients with suspected resectable HCCs were evaluated with CT arteriography before surgery. The results of CT arteriography were compared with those of US, of CT with intravenous contrast medium and of angiography; on the rule, all exams had been performed some days earlier. The diagnosis of HCC was confirmed by US-guided fine-needle biopsy. CT arteriography demonstrated liver lesions in 11 patients. The lesions were hyperdense in 3/11 patients (27.3%) and hypodense and surrounded by a hyperdense ring in 8/11 patients (72.7%). In 4 of 11 patients (36.4%) CT arteriography identified additional tumor nodules and thus surgery was excluded. In the latter cases, on the basis of CT arteriographic findings, US, CT with i.v. contrast medium and angiography were repeated but failed to demonstrate the additional nodules, either because they were too small or because of cirrhotic changes in liver parenchyma. Therefore, CT arteriography is recommended in the evaluation of selected patients, especially when detailed information on liver parenchyma is needed--e.g., before surgery. In these patients CT arteriography can be performed together with preoperative angiography. PMID- 8248587 TI - [Computed tomography evaluation of urothelial carcinomas of the upper urinary tract]. AB - Computed Tomography (CT) is an indispensable noninvasive method for staging transitional cell carcinomas of renal pelvis and ureter. Twenty-seven patients with upper urinary tract tumors were examined and the CT results correlated with histopathologic findings. These tumors exhibit three different development patterns: in our series we identified 17 intraluminal sessile lesions, 6 focal or concentric wall thickenings and finally 7 infiltrating masses; soft-tissue density lesions exhibited in all cases mild contrast enhancement (mean density increase: 38 HU). Even though the attenuation values of tumors do not allow the accurate demonstration of the depth of renal pelvis and ureteral wall invasion, CT can differentiate the lesions which are still confined within the external wall layers (CT1, 17 cases) from those invading the peripelvic-ureteral fat (CT2, 9 cases) or spreading to other organs as well as distant metastases (CT3, 4 cases). Delayed scans can also demonstrate intraluminal lesion extent and sometimes changes of patients' position allow better differentiation of wall outlines from peripheral fat. The overall staging accuracy of CT was 76.66% and its sensitivity was 93.33%. The assessment of lymph node metastases was the major cause of error in the CT evaluation of these tumors. PMID- 8248588 TI - [Abdominal Doppler ultrasonography in the diagnosis of renovascular diseases. Double-blind prospective study]. AB - Our work was aimed at evaluating the sensitivity and specificity and the positive and negative predictive values of abdominal Doppler US in the diagnosis of renovascular disease. Fifty hypertensive patients (23 men and 27 women) with 100 bilaterally single renal arteries (66 of them normal and 34 stenosed) were studied by means of color (26 cases) and duplex (24 cases) Doppler US. The following US variables were considered in the diagnosis of renal artery stenosis: increased peak systolic velocity in the renal artery, increased ratio between peak systolic velocity in the renal artery and in the aorta, increased parenchymal acceleration time and increased resistive index. Feasibility was 100% for all variables. The general multivariate statistic model of discriminate analysis was used to define three calculation levels, according to: a) duplex vs. color Doppler equipment, b) hemodynamic variables and c) optimal cutoff values. US examinations were performed by a single observer in a double-blind study, before angiography, so as to eliminate interobserver variability. On the whole abdominal Doppler US exhibited optimal specificity and fairly good sensitivity. Specificity increased (94% to 97%) with the use of color Doppler according to the different combinations of variables, but sensitivity remained the same. The calculation including all the Doppler variables increased sensitivity from 73% (with true values) to 93% (with threshold values). The positive predictive value demonstrated that 90% of the renal arteries with increased peak systolic velocity and 100% of those with increased parenchymal acceleration time were stenosed. However, the negative predictive value demonstrated that 15% of the renal arteries with no increase in peak systolic velocity is misdiagnosed as normal. In conclusion, abdominal Doppler US can be used to study renovascular disease patients, but the examination must always be based on different hemodynamic variables and true values. PMID- 8248589 TI - [Percutaneous transluminal renal angioplasty. Italian experience at 13 centers]. AB - The data relative to 13 years' experience with percutaneous transluminal renal angioplasty (PTRA) in the 13 major interventional radiology centers in Italy were collected and analyzed to evaluate technical and clinical results. Our aim was to collect homogeneous results in a large series of patients, evaluating both the technical and the clinical success with a long-term follow-up (1 month-13 years). One thousand three hundred forty seven PTRA procedures (including 42 restenoses) in 1,073 patients were retrospectively reviewed. In 807 cases the cause of stenosis was atherosclerosis, in 442 cases fibromuscular dysplasia and in 24 cases arteritis in 12 cases, stenoses were present in patients that had undergone a surgical by-pass. Sixty-two stenoses in patients with solitary kidney and 102 in patients with renal failure were studied separately. The technical success (based on the morphology of the dilated tract) obtained in 91% of cases was considered, together with the clinical success (in 81% of cases), based on the decrease in blood pressure evaluated according to Martin's classification. The blood pressure values collected after the maneuver were also evaluated separately for the different types of stenosis etiology: atherosclerosis, fibromuscular dysplasia or arteritis; moreover, blood pressure was studied in solitary kidney patients and in those with renal failure. Complications were classified as major (4.2%) and minor (4.9%). The high success rate and the low incidence of complications we observed in our series suggest PTRA as the procedure of choice for high blood pressure patients with renal artery stenosis > 50% of the normal caliber. PMID- 8248590 TI - [Arterial recanalization with the Kensey catheter. Preliminary results]. AB - Fifteen patients with chronic obstruction of leg arteries (mean length: 6 cm) were treated with the Kensey catheter and then with PTA. In 11 cases the procedure was successful (74%) and in 2 it was interrupted--in one case for dissection and in the other for vessel perforation. Two maneuvers, after dissection, were ended with PTA and stent placement (overall success rate: 87%). Emergency surgery was never needed. In the cases solved with the Kensey catheter, the cumulative patency rate was 51% at 6, 12 and 24 months. The Kensey catheter did not improve the results of conventional PTA, which means its role is complementary to that of PTA and an alternative to more expensive laser systems, if PTA fails. PMID- 8248591 TI - Absorbed dose from contaminant electrons inside and outside megavoltage photon beams. AB - Megavoltage photon beams interactions with any object on their path give rise to secondary photons and electrons. This fact is particularly important in Radiotherapy. These secondary particles produce an unwanted dose contribution both on beam path and outside the geometrical edges of the irradiation field. The goal of this report is to investigate the dose contribution due to secondary electrons. Ionization measurements were made in a polystyrene phantom by means of a Markus ionization chamber, with X-ray beams produced by a "SL5" Philips and by a "Saturne 43" GE CGR linear accelerators. Specific fields for nasopharynx treatment and for supra-diaphragmatic lymphatic chains treatment ("mantle" fields) were investigated. Both depth ionization values on beam path and depth and surface ionization values outside the geometrical edges of the field were obtained. These measurements were made with and without specific electron filters for 6, 15, 25 MV X-ray beams. The results obtained prove that, both on the beam path and outside the geometrical borders of the field, electron contamination reduction by means of specific filters is important only for lower energy X-ray beams. For this sort of beams, the percentage reduction of electron contamination was found. Moreover, the amount of electron contamination outside the field geometrical edge versus field size was evaluated. PMID- 8248592 TI - [Human errors in the calculation of the Monitor Unit in radiotherapy]. AB - Human mistakes are a major source of error from the definition to the execution of a treatment planning. An unrevealed serious human error can cause therapy to fail therefore it is of fundamental importance to eliminate serious human errors during MU calculation. In this work the human error incidence in MU calculation is evaluated by an independent check. The investigation refers to 1,926 controls collected in about 18 months. Serious daily errors rate (errors causing a 5%, or higher, discrepancy on the daily reference dose) was 1.4% (27/1926). Serious global errors incidence (errors causing a 5%, or higher, discrepancy on the total reference dose) was 0.9% (15/1,731). The data show that the human error in MU calculation is not negligible; they also indicate the value of the independent control of MU, which is an important tool for quality assurance in radiotherapy (like in vivo dosimetry and portal imaging. PMID- 8248593 TI - [Report of a case of cerebellar hemangioblastoma. Description of a case]. PMID- 8248594 TI - [Diffuse micrometastatic involvement of the bone marrow: scintigraphic diagnosis with 201Thallium-chloride. Description of a case]. PMID- 8248595 TI - [Articulate osteomas of the mandibular condyle. 2 cases]. PMID- 8248596 TI - [Sonographic diagnosis of gastric phytobezoar. Description of a case]. PMID- 8248597 TI - [Sonographic diagnosis of the neoplastic course of procelain gallbladder]. PMID- 8248598 TI - [Splenic hemangioma: functional study with double radiotracer single photon emission tomography and technique of sequential marking]. PMID- 8248599 TI - [Multiple leiomyomas in AIDS. Description of a case]. PMID- 8248600 TI - [Sonographic picture, with computed tomography and magnetic resonance in 3 cases of "retrohepatic" gallbladder]. PMID- 8248602 TI - [Barbarisms, acronyms, useless neologisms]. PMID- 8248601 TI - [Holt-Oram syndrome. Description of a case]. PMID- 8248603 TI - [Specification. Combined radio-chemo-surgical treatment of non N2 non-small-cell lung carcinoma]. PMID- 8248604 TI - [Transvaginal ultrasonography in the study of the rectum: combined methods]. PMID- 8248605 TI - [Continuous monitoring of hemoglobin oxygen saturation in mixed venous blood during thoracic anesthesia with selective single-lung ventilation]. AB - OBJECTIVES: The lateral decubitus position, the thoracotomy and selective single lung ventilation all cause serious changes in oxygenation. The aim of this study is to examine the usefulness of continuous monitoring of mixed venous saturation (SvO2) under these conditions. MATERIAL AND METHODS: Twenty-three patients scheduled for pulmonary resection by thoracotomy in lateral decubitus position with single-lung ventilation were monitored continuously for SvO2 by reflection oximetry and a fiber-optic catheter in the pulmonary artery (Abbot Oximetrix 3). Cardiorespiratory profiles were obtained 15 minutes after induction (M1), after 15 minutes in lateral decubitus position (M2), 5 min after start of pleurotomy (M3), 5 min (M4) and 20 min (M5) after lung collapse, 5 min after closure of the thoracic wall (M6), and after 5 min in supine decubitus position (M7). RESULTS: The decrease in SvO2 was not statistically significant. It reached its lowest level at M5 and rose significantly in M6 (p < 0.05). For all SvO2 measurements there was a significant correlation between Oximetrix 3 and Co-oximeter readings. C(a-v)O2 and O2 uptake correlated significantly at all measurements, as did those of PvO2 and Qva/Qt except at M6. VO2I correlated inversely from M1 to M5, and CI an DO2I only at M6 and M7. PaO2 and SaO2 were significantly correlated during single-lung ventilation (M4 and M5) and later (M6). The combined changes in SaO2(a-v) and CI explain the changes in SvO2 only during and after selective ventilation, and the changes are more likely to be reflected by SaO2 than by CI. CONCLUSIONS: Under the conditions studied, continuous monitoring of SvO2 is a reliable method, indicating primarily oxygenation, and cardiac output secondarily, particularly during single-lung ventilation, and also reflecting the balance of oxygen supply and demand in tissues. Given the lack of specificity, however, hemodynamic and gas measurements must be taken to diagnose the cause of any change in the supply/demand balance whenever there is a sudden drop in SvO2. PMID- 8248606 TI - [Anesthesia recovery, gas exchange and postoperative hepatic and renal function in patients with morbid obesity undergoing bariatric surgery: comparison of the effects of halothane, isoflurane and fentanyl]. AB - OBJECTIVES: To compare the postoperative effects of three anesthetic agents, fentanyl, halothane and isoflurane, on recovery from anesthesia, changes in arterial blood gases, and tests of liver and kidney function in morbidly obese patients recovering from vertical ring gastroplasty. MATERIAL AND METHODS: Thirty three patients were studied, randomly distributed into three groups of 11. Induction for all was with atracurium (5 mg), 2.5% thiopentone sodium (5-6 mg.kg 1), succinylcholine (1.5 mg.kg-1) and orotracheal intubation. Anesthesia was maintained with intermittent doses of fentanyl (group F), 2% halothane (group H) or 2.5% isoflurane (group I). All patients received a 50% O2/N2O mixture at a minute volume calculated on ideal weight. Muscle relaxation was achieved by continuous perfusion of atracurium. Postoperative analgesia was by morphine chloride through a lumbar epidural catheter. Time of eye opening and time of extubation were recorded. Arterial blood gas measurements were taken and the results of liver and kidney function tests were recorded until the 7th day after surgery. RESULTS: Eye opening after awakening was earlier in the fentanyl group (6 +/- 5 min), but no differences were found for time of extubation. Blood gas measurements for the 33 patients revealed a significant decrease in PaO2 (58 +/- 14 mmHg), a slight increase of PaCO2 (40 +/- 6 mmHg) and a lower pH (7.32 +/- 0.04) immediately after surgery. On day seven, PaO2 had not yet reached preoperative levels (p < 0.01). These results were independent of anesthetic agent used. Kidney function tests showed significant rises in SGOT (81 +/- 36 U/l), SGPT (150 +/- 110 U/l) and bilirubin (Bil: 15 +/- 5 mmol/l) and decreases in prothrombin activity (PT: 73 +/- 11%) 24 hours after surgery, with later normalization. Urea fell significantly throughout the seven-day period (3.2 +/- 1.3 mmol/l). These results were also independent of the anesthetic agent used. CONCLUSIONS: Morbidly obese patients undergoing gastroplasty recover from anesthesia in the same way regardless of the agent used. The early postoperative period is characterized by severe hypoxemia and transitory changes in kidney function tests. Neither of these findings is dependent on the agent used. PMID- 8248607 TI - [Anesthesia in surgery for epilepsy]. AB - OBJECTIVES: To evaluate the efficacy of general anesthesia during epileptic surgery. MATERIAL AND METHODS: A retrospective study of 64 patients who received general anesthesia during epileptic surgery. In the preoperative period, anticonvulsive medication was adjusted in accordance with plasma levels and withdrawn entirely 8 hours before surgery. After premedication with droperidol and fentanyl, a balanced anesthetic technique was applied, based on pentothal, pancuronium (or vecuronium), fentanyl, N2O and isoflurane. Continuous monitoring of ECG, arterial blood pressure, pulse oximetry, ET CO2 and neuromuscular function. Isoflurane was stopped for 10 min after the opening of the duramadre so that ECoG could be recorded and methohexital or propofol was given in some cases in order to activate the epileptogenic focus. Muscular relaxation was restored intraoperatively following the study of somatosensory evoked potentials. Immediate and later complications related to anesthesia or surgery were recorded. RESULTS: The surgical procedure performed in most cases was temporal or frontal resection, with a mean duration for anesthesia of 377 +/- 50 min and for surgery of 318 +/- 50 min. Only one patient received local anesthesia and no hemodynamic changes were observed. Perioperative complications were cerebral edema (4 cases), arrhythmia (2 cases) and bronchospasm (1 case). Postoperative complications were as follows: 3 of 9 patients undergoing callosotomy required mechanical ventilation for 24 hours, 4 patients experienced language alterations, 3 wounds were infected, 2 cases of hemiplegia were observed, 1 status epilepticus occurred after administration of propofol and there was 1 case of respiratory distress. Anticonvulsive medication was given parenterally after surgery. CONCLUSIONS: General anesthesia is a safe and effective method for epileptic surgery, with local anesthesia providing additional sedation for isolated cases. Appropriate treatment requires an understanding of the pharmacokinetics and pharmacodynamics of the drugs used, as well as knowledge of the condition and the anticonvulsive medications used. PMID- 8248608 TI - [Combined subarachnoid and epidural block with a single injection, with a modified Tuohy needle and used in hip surgery]. AB - OBJECTIVES: To evaluate prospectively the advantages and disadvantages of a combined subarachnoid and epidural block with a modified Tuohy needle during hip surgery. MATERIAL AND METHODS: Fifty ASA II-III patients undergoing full hip arthroplasty were studied. The same technique was used with all patients; 2 ml of 0.5% hyperbaric bupivacaine (10 mg) was used for the subarachnoid block. Ten ml of 2% lidocaine (200 mg) was injected by epidural catheter into 25 patients (group I); 5 ml of 0.5% bupivacaine (25 mg) was injected into the others (group II). A Tuohy 17 G needle with a Huber point and supplemental orifice for a 26 G intradural needle was used, along with an epidural catheter. The level of difficulty of the technique was evaluated, as were unusual events, the metameric level of sensory block reached, and intraoperative (hypotension, bradycardia or both) and postoperative (head or lower back pain) complications. RESULTS: The level of difficulty was nil in 60% of the cases, average in 26% and high in 14%. The quality of the motor-sensory blockade was excellent and the thoracic level of sensory blockade (mean +/- SD) was 6.2 +/- 1.4 for group I and 8.2 +/- 1.1 for group II. The difference between the two groups was significant (p < 0.05). Hypotension was found in 6% of all patients, bradycardia in 20% and both in 14%, with no significant differences between the two groups. No case of headache was reported and the level of patient satisfaction with the anesthesia was excellent in 92% of the cases. CONCLUSIONS: A combined subarachnoid (10 mg hyperbaric bupivacaine) and epidural (200 mg lidocaine or 25 mg bupivacaine) blockade provides an excellent sensory block with few side effects. The technique facilitates epidural treatment of postoperative pain. PMID- 8248609 TI - [EMLA. A new topical anesthetic]. AB - The method most widely used at present for cutaneous analgesia is local anesthetic infiltration. Pain occurs when the needle penetrates and when the anesthetic is deposited and diffuses. The procedure is not risk-free. It is therefore not surprising that many attempts have been made to develop an anesthetic formula that is effective when applied topically to intact skin. Early efforts were unsuccessful due to either low analgesic efficacy or side effects. The problem was solved several years ago in the European market by the EMLA (Eutetic Mixture of Local Anesthetics) cream. The EMLA cream has been shown to be useful as a topical anesthetic on either intact or damaged skin. Its clinical profile is safe, with a low incidence of side effects. We feel it is a useful tool for preventing pain in a variety of procedures that call for cutaneous analgesia. It may also be used as premedication for perineural injection of anesthetics, providing greater comfort for the patient. This article describes the main pharmacological principles that make EMLA an effective topical analgesic, as well as indications for its use in the clinical practice of anesthesiology. PMID- 8248610 TI - [E. Pi y Molist's and J.A. Reynes' memoirs on chloroform (1849): 2 forgotten classics of the Spanish bibliography on anesthesia]. AB - This paper reviews the circumstances surrounding the first prize given in Barcelona in 1849 for a report on anesthesia. The two winning studies by the Barcelona physicians Emilio Pi y Molist and Jose Antonio Reynes are summarized, evaluated and compared with other similar contemporary works. PMID- 8248611 TI - [Propofol and retrobulbar anesthesia for cataract extraction]. AB - We divided 80 patients undergoing cataract surgery into two groups of 40: one control group and one propofol group to whom 1-1.5 mg/kg of propofol was administered before retrobulbar anesthesia. The following parameters were recorded before and after retrobulbar anesthesia: systolic and diastolic arterial pressure (SAP and DAP), heart rate (HR) and finally arterial oxygen saturation through pulse oximetry. Pain was also measured on the Scott-Huskisson visual analog scale. For patients in the control group a rise in arterial pressure over baseline values after 5 minutes (p < 0.01) was observed, while a decrease was found in the propofol group (p < 0.01). The rise after 5 minutes in the control group was significant when compared with the measurements for the propofol group (p < 0.01). The pain measure for the control group reached 5.53 +/- 1.54 on the Scott-Huskisson scale, but was 0 in the propofol group. Measurements on the pain scale correlated positively with diastolic arterial pressure 5 minutes after blockade in the control group (p < 0.05). The technique studied affords greater comfort for the patient, presents no special difficulties for the anesthesiologist performing the retrobulbar blockade, and causes no complications. PMID- 8248612 TI - [Vascular lesions in surgery of the lumbar spine]. AB - We present two cases of iliac artery damage arising from surgery to correct a lumbar disc hernia due to ventral perforation. The first was a pseudoaneurysm of the left iliac artery with a retroperitoneal hematoma diagnosed by ultrasound and TAC on the third day after surgery. The second was a retroperitoneal hematoma diagnosed by intraoperative ultrasound after the patient was shifted to a new position. Emergency laparotomy was performed on both patients to repair the damage. This is an unusual but serious complication, and in most cases requires intuitive diagnosis and lifesaving surgery. PMID- 8248613 TI - [The use of the spinal route in the treatment of chronic pain of oncologic origin]. PMID- 8248614 TI - [Transient blindness during transurethral resection of the prostate: is glycine the culprit?]. PMID- 8248615 TI - [Anesthesia in variegate porphyria]. PMID- 8248616 TI - [Late extrapyramidal syndrome with low doses of droperidol]. PMID- 8248617 TI - [Anesthesia in liver and kidney transplantation]. PMID- 8248618 TI - [Subcutaneous emphysema without pneumothorax after difficult intubation]. PMID- 8248619 TI - [More on amylene]. PMID- 8248620 TI - [Drug surveillance in anesthesia]. PMID- 8248621 TI - [Hyperthyroidism: preoperative control with diltiazem]. PMID- 8248622 TI - Immunological localization of Bacillus thuringiensis serovar israelensis toxins in midgut cells of intoxicated Anopheles gambiae larvae (Diptera: Culicidae). AB - Fourth instar larvae of Anopheles gambiae were intoxicated with doses of purified crystals from Bacillus thuringiensis serovar israelensis (Bti) corresponding to 50-fold the LC50 after 24 h. Midguts were dissected after various contact times, then processed for immuno-light and -electron microscopy. Immunodetection on thin sections was performed using affinity-purified rabbit IgG against Bti crystal CryIVD or CytA polypeptides, in combination with anti-rabbit IgG/peroxidase. Both polypeptides were detected by optical and electron microscopy after 15 min of contact with Bti crystals on the apical brush border of midgut cells, but only in the gastric caeca and posterior stomach. No specific signal was detected in the other parts of the midgut, i.e. the cardia cells and the anterior stomach. These results confirm that mosquito midgut cells are the primary target for the toxins and that binding to specific receptors on the apical microvilli membrane is the initial step of delta-endotoxin action. PMID- 8248623 TI - Mutations in phoB, the positive gene activator of the pho regulon in Escherichia coli, affect the carbohydrate phenotype on MacConkey indicator plates. AB - Mutants defective in phoB, the positive gene activator of the Escherichia coli pho regulon, exhibit aberrant behaviour on MacConkey indicator plates. They appear pale in the presence of a fermentable carbon source such as trehalose, maltose or glucose. The addition of at least 5 mM phosphate corrects this defect. Colonies of phoB+ strains turn red on MacConkey indicator plates and derepress the pho regulon when the cells are able to ferment the carbon source. In contrast, the inability to ferment the carbon source maintains the pho regulon in the repressed state. PMID- 8248624 TI - Altered expression of several genes in IIIManL-defective mutants of Streptococcus salivarius demonstrated by two-dimensional gel electrophoresis of cytoplasmic proteins. AB - Mannose, glucose and fructose are transported in Streptococcus salivarius by a phosphoenolpyruvate:mannose phosphotransferase system (PTS) which consists of a membrane-bound Enzyme II (EII) and two forms of IIIMan having molecular weights of 38,900 (IIIManH) and 35,200 (IIIManL), respectively. We have previously reported the isolation of spontaneous mutants lacking IIIManL and showed that they exhibit higher beta-galactosidase activity than the parental strain after growth on glucose, and that some of them constitutively express a fructose PTS which is induced by fructose in the parental strain. In an attempt to determine whether the expression of other genes is affected by the mutation and what the physiological link is between them, we examined three S. salivarius IIIManL defective mutants (strains A37, B31 and G29) and the parental strain using two dimensional gel electrophoresis after growth of the cells on a variety of sugars. After growth on glucose, five new proteins were detected in the cytoplasm of the three mutants. Two of these proteins were induced in the parental strain by galactose or oligosaccharides containing galactose, and one was specifically induced by melibiose. The other two proteins were not detected in the parental strain under any of the growth conditions tested. Two other proteins were only detected in glucose-grown cells of mutant A37, and a protein associated with the metabolism of fructose was constitutively expressed in mutants B31 and G29. Moreover, we have found that under identical growth conditions the amounts of several other proteins which were detected in the parental strain were either increased or decreased in the mutants. Globally, our results have indicated that (1) the expression of several genes was affected in the spontaneous IIIManL defective mutants; (2) some of the proteins abnormally produced in the mutants were specifically induced in the parental strain by sugars; (3) the phenotypic modifications observed in the mutants were of two types: most were observed solely after growth of the cells on glucose whereas the others were glucose independent; and (4) the mutants shared common phenotypic traits, but also exhibited idiosyncratic characteristics. PMID- 8248625 TI - Proteolytic and oxidoreductase activity of Treponema denticola ATCC 35405 grown in an aerobic and anaerobic gaseous environment. AB - The cells of a human oral spirochete, Treponema denticola ATCC 35405, and of seven clinical isolates of this organism obtained from the subgingival dental plaque of periodontitis patients were studied for their ability to grow in an aerobic and an anaerobic environment, and for their profile of peptidohydrolase and oxidoreductase enzymes. The growth yield of aerobically grown cultures was either comparable to or higher than that of anaerobically grown ones regardless of whether prereduced broth, freshly prepared broth or oxidized broth was used. However, elimination of certain supplements from the growth media resulted in poor growth regardless of the nature of the gaseous environment. The microscopic morphology and motility of the cells were not affected by differences in the gaseous atmosphere. Quantitative studies on several peptidohydrolase activities suggest that anaerobically grown cells displayed higher specific activity especially toward N alpha-L-prolyl-2-naphthylamine, indicating that increased synthesis of proline iminopeptidase enzymes (or enzyme) of the cells was associated with anaerobic growth conditions. The formation of enzymes hydrolysing N alpha-benzoyl-DL-arginyl-2-naphthylamine (and the corresponding p-nitroaniline) was not affected to the same extent. Growth experiments suggest that T. denticola ATCC 35405 is a facultatively anaerobic spirochete instead of an obligate anaerobe as reported in previous literature. The quantitative enzyme studies suggest that the gaseous growth atmosphere of the cells can exert a selective effect on the activity levels of certain peptidolytic enzymes of this organism. Such effects were not observed when the whole cells were studied by means of qualitative or semi-quantitative enzyme tests. The activities of catalase, peroxidase and superoxide dismutase of the cells were low and variable. Because of this, it was not possible to relate these oxidoreductase activities to the composition of the gaseous atmosphere. PMID- 8248626 TI - Measurement of the net production of acidity by a sulphate-reducing bacterium: experimental checking of theoretical models of microbially influenced corrosion. AB - The net production and consumption of acidity by Desulfovibrio fructosovorans, growing with lactate as the carbon and energy source, were measured in an unbuffered medium in a pH-controlled bioreactor. At alkaline pH (7.2 and 8.5), net acidity production was measured. At pH 6.0, acidity consumption was obtained, although bacterial growth was not observed. These observations are in good agreement with theoretical predictions emphasizing the key role of H+ ions in the relationship between the metabolism of sulphate-reducing bacteria and microbially influenced corrosion. PMID- 8248627 TI - A diverse transketolase family that includes the RecP protein of Streptococcus pneumoniae, a protein implicated in genetic recombination. PMID- 8248628 TI - Screening of a cosmid library of Mycobacterium bovis BCG in Mycobacterium smegmatis for novel T-cell stimulatory antigens. AB - We have developed a novel method for screening a Mycobacterium bovis (BCG) cosmid library in Mycobacterium smegmatis for the detection of immunostimulatory T-cell antigens (Ag). Distinctive protein banding patterns were demonstrated in culture filtrates of three of 30 recombinant M. smegmatis clones: pBCCS13 (41 and 73 kDa); pBCCS221 (30, 50 and 68 kDa); pBCCS223 (100 kDa). Western immunoblots indicated that monoclonal antibodies (mAb) directed to the previously characterized 19-, 30-, 38-, 65- and 71-kDa mycobacterial Ag were not reactive with the distinctive recombinant proteins. Furthermore, T-cell Western blots demonstrated that fractions containing the distinctive proteins were immunostimulatory. A given tuberculin-positive donor expressed unique patterns of blastogenic reactivity to protein fractions isolated from each of the three recombinant clones. Restriction enzyme digests of the three recombinant BCG inserts revealed distinctive DNA-banding patterns. The immunostimulatory Ag, therefore, are most likely encoded within different regions of the BCG genome, as contained within three distinct inserts. T-cell Western blots further indicated a heterogeneity in the repertoire of BCG-responsive T cells since tuberculin positive donors varied in the pattern of reactivity to protein fractions isolated from the same recombinant filtrate. Most likely, immunity to M. tuberculosis results from activation of a heterogeneous array of T cells targeted to multiple immunostimulatory Ag. The method we describe should greatly enhance our ability to define the full spectrum of T-cell Ag encoded by mycobacteria, particularly those which are secreted proteins. PMID- 8248629 TI - Identification of six open reading frames in the Salmonella enterica subsp. enterica ser. Typhi viaB locus involved in Vi antigen production. PMID- 8248630 TI - Factors affecting reproducibility of random amplified polymorphic DNA fingerprinting. AB - The reproducibility of random amplified polymorphic DNA (RAPD) was tested using two different thermal cyclers and three brands of Taq DNA polymerase. Three different oligonucleotides were used to obtain patterns of amplified fragments from three DNA samples (Escherichia coli, Bacillus subtilis, and Thermococcus littoralis). Experiments were repeated three to six times. Apart from the expected between-oligonucleotide and between-DNA variations, between-thermal cycler and between-DNA polymerase variations were observed. Within the DNA oligonucleotide-DNA-polymerase-thermal cycler, reproducibility was excellent when the thermal cycler equipped with the best temperature regulation was used, but was not as good with another brand of thermal cycler. PMID- 8248631 TI - Biotyping of Streptococcus thermophilus strains by DNA fingerprinting. AB - DNA of 33 strains of Streptococcus thermophilus (7 isolated from natural whey cultures utilized as starter, 25 from commercial yogurts and 1 reference strain) was analysed by restriction endonuclease digestion and both conventional and pulsed-field agarose gel electrophoresis. The restriction patterns after digestion of total DNA with HindIII, EcoRI and BamHI enabled 10 different groups to be distinguished by conventional electrophoresis. Digesting genomic DNA with SmaI or NotI and separating it by pulsed-field gel electrophoresis, the 33 strains could be classified into 5 groups. The same result was obtained after digestion with HaeIII and conventional electrophoresis. The 33 isolates could be divided into 13 groups when diversities among patterns obtained after both the first and second method were considered. A combination of the two techniques turned out to be the most reliable methodological approach for characterizing strains of S. thermophilus for scientific, industrial and legal purposes. PMID- 8248632 TI - Genetic studies of cefotaxime resistance in Streptococcus pneumoniae: relationship to transformation deficiency. AB - A laboratory pneumococcal strain resistant to cefotaxime was studied by DNA induced transformation in order to characterize its genetic structure. At least three independent genes were required to confer the highest level of resistance to this beta-lactam antibiotic. The accumulation of mutations in these three genes accounted for three levels of resistance. Mutation of the gene encoding penicillin-binding protein 2x was very likely responsible for the first step of resistance, which was a prerequisite for sequential increase in resistance. Additionally, strains highly resistant to cefotaxime were defective for natural transformation. Revertants of these strains were frequently observed. Such strains had recovered full transformability, suggesting a correlation between the inability to be transformed and a high level of resistance to cefotaxime. The possibility of electrotransforming these highly resistant strains suggests that natural transformation is probably blocked at the DNA-uptake level. PMID- 8248633 TI - Inclusion bodies and crystals of Bacillus sphaericus mosquitocidal proteins expressed in various bacterial hosts. AB - Mosquitocidal proteins of 42 and 51 kDa from Bacillus sphaericus were produced in acrystallogenic B. sphaericus and B. subtilis strains. In both species, transformants containing each protein expressed individually were non-toxic for mosquito larvae and produced small inclusions which did not have a crystalline ultrastructure. When both components of the binary toxin were expressed together, toxicity was restored: oval and round amorphous inclusions were produced in B. subtilis, and typical native-type crystals were synthesized in B. sphaericus. In B. subtilis, native-type crystals were produced only when the two components of the binary toxin were synthesized as a 93-kDa fusion protein. PMID- 8248634 TI - [Preliminary study of HTLV-I seroprevalence in Chilean Indian populations]. AB - Aiming to seek the origin and define the prevalence of HTLV-1 infections, 464 blood samples from aboriginal populations proceeding from isolated regions of the north and south of Chile were studied. Antibodies against HTLV were measured with agglutination tests and confirmed with immuno-fluorescence and Western Blotting. Seven out of 107 (6.5%) blood samples from Atacama indians, 2 out of 202 (1%) from Mapuche indians and 3 out of 155 (1.9%) from Huilliche aborigines were positive. These results highlight an important presence of the virus in indigenous populations, specially in the extremes of the country. These findings could suggest an indigenous (mongoloid) origin of HTLV-1 in Chile, specially in Chiloe, where apparently there has been no contact with african or japanese populations, that could be the origin of the contamination. PMID- 8248635 TI - [Respiratory symptoms and pulmonary function in O' Higgins antarctic base residents]. AB - The aim was to study pulmonary function and physical capacity and their relation to respiratory symptoms among military personnel before and after one year of permanence in an Antarctic base. In 21 men aged 38 +/- 5.5 years, enquiries about smoking habits and respiratory symptoms, spirometry, bronchial provocation test and measures of aerobic capacity and peak expiratory flow were performed. After 75 days of permanence in the base, there was a significant increase in weight, exertional dyspnea appeared in 8 subjects and there was an 8.4% decrease in aerobic capacity. The bronchial provocation test was positive outdoors in 7 of the 21 staff members; when performed indoors, no subject had a positive test and in Santiago only one subject had a positive test (Chi sq p < 0.025). After 7.5 and 11 months of permanence, forced vital capacity decreased in 13% and no diurnal differences of peak expiratory flow were observed. In conclusion, the permanence in Antarctica may slightly affect the respiratory system with the appearance of exertional dyspnea and outdoor airway hyper reactivity in some residents. PMID- 8248636 TI - [Treatment of mild hypertension with captopril and captopril with hydrochlorothiazide. Impact on blood pressure values and quality of life]. AB - The effects of captopril alone or associated to hydrochlorothiazide on blood pressure and quality of life was studied in 244 hypertensive subjects. After a medication wash out period of 30 days (and 7 additional days when diuretics were used) patients were treated initially with captopril 25 mg b.i.d. (scheme A); if blood pressure was not normalized, hydrochlorothiazide 25 mg o.d. was added (scheme B) and the dose of captopril was increased to 50 mg bid (scheme C) in a stepwise fashion. Sixteen patients (6.6%) were lost from follow up; in the rest a significant reduction of blood pressure was obtained at the end of the treatment period (from 163/98.9 +/- 19.7/3.1 to 143/85 +/- 12.7/6.7 p < 0.05), without changes in cardiac frequency. Using captopril alone, blood pressure was normalized at 15 and 75 days in 62.9 and 70.6% of patients respectively. With captopril plus hydrochlorothiazide blood pressure was normalized in 80.9% of patients. Quality of life improved significantly from of 42.8 +/- 1.7 to 50.7 +/- 15 score points (p < 0.05) secondary effects were observed in 7.4% of patients. It is concluded that captopril alone or associated to hydrochlorothiazide is effective in the treatment of mild hypertension, produces few secondary effects and improves the quality of life. PMID- 8248637 TI - [Rapidly progressive glomerulonephritis without immune deposits: detection of neutrophil anticytoplasmic antibodies]. AB - Antineutrophil cytoplasmic autoantibodies (ANCA), are serologic markers of disease in rapidly progressive glomerulonephritis without immune deposits (pauci immune) and vasculitis, and could play a pathogenic role in these diseases. We communicate 5 patients with pauci-immune rapidly progressive glomerulonephritis; four of them in the evolution of necrotizing systemic vasculitis (Wegener's granulomatosis and microscopic polyarteritis) and one case with lesions limited to the kidney. All of them were associated with the presence of ANCA. The immunofluorescence pattern (cytoplasmic and perinuclear) and the antigenic specificity of ELISA assay, [antiproteinase-3 (PR-3) and antimyeloperoxidase (MOP)], are useful laboratory tools for the diagnosis and clinical management of these patients. PMID- 8248638 TI - [Changes in the clinical presentation of primary hyperparathyroidism. Analysis of 84 cases]. AB - During the last two decades, the clinical presentation of primary hyperparathyroidism (PHP) has changed due to the routine use of multiphasic biochemical screening tests. We assessed 84 patients with PHP treated in our service between 1977 and 1991. The yearly incidence increased from 1.6 to 7.6 patients/year with the introduction of multiphasic biochemical testing in our hospital in 1982; likewise the proportion of asymptomatic patients increased from 12.5 to 40.7%. The most frequent presenting symptoms were bone pain and renal colic. Nineteen percent of patients were over 70 years old and this age group had distinct clinical features. The plasma chlorine/phosphorus ratio was abnormal in 95% of cases; on the contrary only 7 of 18 patients had a urinary calcium excretion over 300 mg/day. Cervical ultrasound, performed in 45 patients had a positive predictive value of 78% to localize the lesion. Bone density was below fracture threshold in 50% of studied patients. The principal surgical finding was the presence of adenoma. Twenty one percent of patients had symptomatic hypocalcemia during the first week after surgery; however, only 2.5% of patients continued to have hypocalcemia one month after surgery. One patient had an inferior laryngeal nerve damage and two a cervical hematoma. It is concluded that the introduction of massive calcium measurements has allowed an early diagnosis of asymptomatic PHP, specially in elderly people. PMID- 8248639 TI - [Hemodynamic effects of dobutamine in critical patients: discrepancies between parameters of tissue oxygenation and lactic acid]. AB - A retrospective study on the effects of dobutamine on hemodynamic parameters of tissue oxygenation and lactic acid levels was performed in 35 patients. The use of the drug in a mean dose of 5.0 +/- 2.1 ug/kg/min was associated with a significant increase in cardiac index, oxygen transport and consumption and decrease in arterial lactic acid levels. A separate analysis of patients with or without basal hyperlactacidemia and those with favorable (decrease) or unfavorable (no change or increase) lactic acid response to the drug, was performed. This analysis disclosed important disagreements between changes in hemodynamic parameters of oxygen consumption and transport and arterial lactic acid levels, as indexes of tissue perfusion. It is concluded that is advisable to monitor together hemodynamics, oxygen transport and consumption and arterial lactic acid in patients with severe cardiorespiratory disturbances or shock. PMID- 8248640 TI - [Etiologic diagnosis of pneumonia in HIV-AIDS patients. Usefulness of fibrobronchoscopy]. AB - AIM: to assess the usefulness of fiberoptic bronchoscopy in the etiologic diagnosis of pulmonary infiltrates in patients with AIDS. PATIENTS AND METHODS: in 25 patients with AIDS and pneumonia, 31 fiberoptic bronchoscopies with bronchoalveolar lavage were performed; in 11 occasions, a transbronchial biopsy was also performed. RESULTS: in 24 of the 31 procedures (77%) an etiologic diagnosis was made. There were 7 false negative results; in two of these patients, a repeated examination allowed an etiologic diagnosis. The etiologies of the infiltrates were pyogenic bacteria in 14 cases. Pneumocystis carinii in 9, mycobacteria in 3, cytomegalovirus in one and a lymphocytic interstitial pneumonia in one. In 5 patients, the association of two or more etiological agents was observed. Except one case, all patients had a good response to specific treatments. There were no complications attributed to the bronchoscopic procedure. CONCLUSIONS: fiberoptic bronchoscopy and bronchoalveolar lavage are safe and effective procedures for the etiologic diagnosis of pulmonary infiltrates in patients with AIDS. PMID- 8248641 TI - [Is it justifiable to use antiplatelet drugs as a universal protection in patients with chronic glomerular damage?]. AB - In many renal diseases, glomerular thrombosis may play an important role in the development of glomerular sclerosis and progression of renal failure. The aim of this study was to assess the effect of antiplatelet agents on the evolution of patients with chronic glomerular disease. Twenty four patients aged 48 +/- 17 years (21 with idiopathic glomerulonephritis, one with systemic lupus erythematosus and two with type II diabetes mellitus) were treated with aspirin and dipyridamole or aspirin alone during 23.9 +/- 17.5 months. The patients were followed during 31.8 +/- 23 months; seven patients had a progressive deterioration of renal failure requiring dialysis or transplantation (Group A) and 17 had a stable or improving renal function (Group B). Initial serum creatinine was significantly higher in group A than in group B (3.6 +/- 1.6 vs 1.5 +/- 1.5 mg/dl respectively p = 0.003); no other significant differences in the initial assessment were observed between both groups. It is concluded that antiplatelet agents may delay the progression of renal disease when started in patients with normal or slightly deteriorated renal function. PMID- 8248642 TI - [Infective endocarditis: clinical aspects and necropsy study in 20 cases]. AB - Aiming to know the principal anatomo-pathological findings in patients that died with a diagnosis of infective endocarditis, the clinical histories and necropsy protocols of such patients that died between 1984 and 1989 were reviewed. Twenty patients (11 male) aged 43.2 +/- 16.1 years were studied. In nine, the endocarditis was subacute, and 80% had a history of valvular or congenital heart disease. The principal clinical complication was sepsis followed by neurological, nephrological and cardiac complications. The necropsy disclosed cardiac, nephrological and abdominal lesions in 100, 80 and 65% of patients respectively. In 65% of necropsies, neurological injuries were found (cerebral edema in 7 patients and hemorrhagic complications in 11). It is concluded that extra cardiac lesions are frequent in patients dying with a diagnosis of infective endocarditis. PMID- 8248643 TI - [Arterial hypertension: therapeutic explorations for an old disease]. PMID- 8248644 TI - [Percutaneous transluminal angioplasty in stenosis of Blalock-Taussig pulmonary systemic shunt in an adult patient]. AB - AIM: The report of a female patient with tricuspid atresia and severe stenosis of the pulmonary artery, stenosis of the left pulmonary-systemic shunt and occlusion of the right systemic-pulmonary shunt. BACKGROUND: The use of transluminal angioplasty has been extended to the treatment of several congenital cardiopathies and, in some of them, it is the therapeutic procedure of choice. METHODS: Dilatation of the distal stenosis of the pulmonary-systemic shunt with a coaxial system composed initially by an 8 French right Judkins carrier catheter with a soft tip through which a 0.014" (0.036 cm) coronary angioplasty guidewire and 3 and 4 mm balloon catheters were advanced successively. Subsequently a 0.028" interchange Rosen guide was advanced, gradually introducing balloon catheters up to 8 mm of diameter. RESULTS: The dilatation with balloon catheter systems of increasing diameter allowed to amplify the stenosis zone, improving arterial oxygen partial pressure and saturation without evidences of left lung hyperflux. Considering the anatomic characteristics of the pulmonary systemic shunt, the use of carrier catheters facilitates the penetration of balloon catheters to the stenosis zone. CONCLUSION: Percutaneous transluminal angioplasty may be used with success in patients with stenosis of Blalock-Taussig shunts, independent of the anastomosis curvature and even if the magnitude of the stenosis is significant. PMID- 8248645 TI - [Budd-Chiari syndrome. Report of 2 surgical cases]. AB - Two female patients with Budd-Chiari syndrome and suffering of Polycythemia Vera are reported. Both patients were operated performing a central spleno renal shunt with preservation of the spleen and ligation of the splenic artery. The patients are well 6 and 24 months after the operation respectively. The principal features and therapeutic alternatives of this condition are reviewed briefly. PMID- 8248646 TI - [Genetic, molecular, and immunologic aspects of Vibrio cholerae infection]. AB - The deterioration of the economical and social conditions of the majority of the population in the Americas the last 20 years has generated several epidemics of enteric infections in the region, dramatically manifested by the current massive and widespread cholera outbreak. The absence of cholera from the continent for more than 100 years, the worsening environmental conditions, the biological peculiarities of Vibrio cholerae El Tor such as decreased virulence, which generates increased number of carriers, and its improved ability to thrive in the environment are probably responsible for the rapid dissemination of the disease through out the continent. Genetic and molecular studies of the biology of V cholerae have permitted identification of a variety of new virulence factors besides the enterotoxin, and are also helping to unravel the exquisite mechanisms that regulate the expression of these virulence factors in response to different stimuli. Molecular studies of V cholerae chromosomal and plasmid DNA, and of chromosomal and plasmid gene products, with techniques such as DNA hybridization and multilocus enzyme analysis are improving the characterization of V cholerae strains, resulting in progress in understanding their epidemiology in different communities. The non-invasive character of V cholerae infections, epidemiological and immunological studies suggest that the disease and current vaccines fail in providing an effective and long lasting immunity, and that the control of the disease in endemic areas by the use of vaccines may therefore be unfeasible. Similar studies indicate that the provision of safe drinking water, adequate sewage disposal, sufficient nutrition, and education remain the most effective measures for controlling the disease. PMID- 8248647 TI - [Scientific research in clinical medicine]. AB - A Faculty of Medicine requires that its academics devote an important part of their duties to scientific research. It does not seem possible to perform university teaching without a personal experience in the methods, techniques and practices of scientific research. Notwithstanding, keeping in mind the fundamental importance of medical practice in the training of professors of a Faculty of Medicine, it is possible to consider a clinical teaching staff, to whom scientific research would not be demanded. This staff would coexist with an academic research staff and would have a limited dedication to clinical research. The object of clinical scientific research is mankind and its purpose relates directly to the medical act. As a consequences the responsible should be a physician. This fact implies that the Faculty of Medicine has the responsibility to create an appropriate environment and system to promote training and research in the clinical area as well as in preclinical and basic areas. A doctorate program in medical sciences in proposed as the most suitable method for the development of clinical scientific research and for the training of young researchers in this area of medical knowledge. PMID- 8248649 TI - [Problems derived from condom commercialization]. PMID- 8248648 TI - [Health, power, and democracy. Notes for a primary care theory]. AB - Health services are relentlessly deteriorating due to the persistent increase in their costs. This deterioration not only affects people's health. This service is directly linked to personal and family security and populations social equity expectatives. Thus, its failure inevitably leads to a gradual loss of State and public authorities legitimacy, threatening the stability of the democratic system. In the long run, it also affects the legitimacy of medical profession. To overcome this problem, considering the limited possibilities of a relatively poor country, a new health services model is proposed, based on a massive strengthening of primary care, organized in a way in which the community assumes an important part of the responsibility for the health of its members. To achieve this purpose the use of the concept of "empowerment" is proposed, practically applied through the "pedagogic investigation-action". As a result a noticeable increase in the general health level of the population is expected. At the same time, one of the serious threatens to the legitimacy of the Democratic State will be eliminated. PMID- 8248650 TI - [Symptomatic profile of the climacteric woman]. PMID- 8248651 TI - [A medical conflict in 1871]. AB - The first medical strife in Chile occurred in the city of Valparaiso. The intendant protested to the examining board of physicians because 5 doctors refused to give emergency medical care to a patient late at night. "Is fair that both the authority and the public have the right to demand those services and it is not natural that their fulfillment depend on the good or bad will of the physicians". The intendant proclaimed a decree establishing a weekly obligatory nocturnal medical services of two physicians under police control and fine threaten. The 14 doctors of the city menaced to resing to their profession considering that the decree "violates constitution and laws." The medical corps of Santiago made common cause with their colleagues "profoundly irritated". The conflict was finally resolved. PMID- 8248652 TI - [Epidemiological evolution of human hydatidosis in Chile, 1965-1988. Impact of livestock programs in the XI and XII regions]. AB - The aim of this work was to analyze the possible impact of livestock echinococcosis control programs in the XI and XII regions of Chile, on the incidence of human echinococcosis. National morbidity was stable until 1984, with an abrupt and important decrease afterwards. Mortality showed a slight tendency to decrease during this period. There was an increase in lethality, that was equivalent and contemporary to the communicated decrease in morbidity after 1985. It is concluded that the national and regional decrease in the communicated cases of echinococcosis is due to a decrease in the notification of the disease. This problem was originated in administrative decisions of the Ministry of Health in 1985, whose effects persist. An instrument to correct retrospectively this low notification is proposed. PMID- 8248653 TI - A case for Bayesianism in clinical trials. AB - This paper describes a Bayesian approach to the design and analysis of clinical trials, and compares it with the frequentist approach. Both approaches address learning under uncertainty. But they are different in a variety of ways. The Bayesian approach is more flexible. For example, accumulating data from a clinical trial can be used to update Bayesian measures, independent of the design of the trial. Frequentist measures are tied to the design, and interim analyses must be planned for frequentist measures to have meaning. Its flexibility makes the Bayesian approach ideal for analysing data from clinical trials. In carrying out a Bayesian analysis for inferring treatment effect, information from the clinical trial and other sources can be combined and used explicitly in drawing conclusions. Bayesians and frequentists address making decisions very differently. For example, when choosing or modifying the design of a clinical trial, Bayesians use all available information, including that which comes from the trial itself. The ability to calculate predictive probabilities for future observations is a distinct advantage of the Bayesian approach to designing clinical trials and other decisions. An important difference between Bayesian and frequentist thinking is the role of randomization. PMID- 8248654 TI - The case for frequentism in clinical trials. PMID- 8248655 TI - The value of randomization and control in clinical trials. AB - This paper examines the two principal justifications that have been offered for the standard conditions that clinical trials be randomized and controlled, with the conclusion that, strictly speaking, neither justification is valid. It is argued, on the other hand, that a Bayesian analysis of clinical trials affords a valid, intuitively plausible rationale for selective controls, and marks out a more limited role for randomization than it is generally accorded. The feasibility of retrospective trials is then considered in the light of these conclusions. PMID- 8248656 TI - The role of p-values in analysing trial results. AB - The current widespread practice of using p-values as the main means of assessing and reporting the results of clinical trials cannot be defended. Reasons for grave concern over the present situation range from the unsatisfactory nature of p-values themselves, their very common misunderstanding by statisticians as well as by clinicians and their serious distorting influence on our perception of the very nature of clinical trials. It is argued, however, that only by fully understanding the reasons why they have become so universally popular can we hope to change opinion and introduce more sensible ways of summarizing and reporting results. Some of the ways in which this might happen are discussed. PMID- 8248657 TI - Statistical and ethical issues in monitoring clinical trials. PMID- 8248658 TI - Clinical trials and causation: Bayesian perspectives. AB - In addition to the safety, it is essential to establish the causal efficacy of extant and new treatments, and well-designed clinical trials are thought by most to be the 'gold standard' to accomplish this. Contrary to most statisticians' and regulators' views, however, I will argue that the concept of causation involved in clinical trials is not all that clear. I discuss the manipulability approach to causation, interpreted counterfactually, which seems to fit causation as it is found in such sciences as physiology, but it has unclear relations to a concept of causation proposed by a number of epidemiologists. I characterize 'epidemiological causation' as probabilistic and formulated at a population level, and dependent on certain general criteria for causation as well as study design considerations. I then attempt to clarify the connections between these concepts of causation and Cartwright's views on complexity and causality, a 'Bayesian' framework proposed by Rubin and further elaborated by Holland, and Glymour and his colleagues' recent directed graphical causal modelling approach. PMID- 8248659 TI - Applying Bayesian ideas in drug development and clinical trials. AB - The Bayesian paradigm emphasizes that studies are not performed in isolation, and that external evidence can be used formally in the design, monitoring and reporting of clinical trials. A variety of tools for assessing the current evidence for treatment efficacy are presented, making use of graphical display to provide insight into ethical and efficiency issues in starting and stopping trials--these are illustrated with a trial in osteosarcoma that is currently taking place. Finally we recommend that an additional 'interpretation' section is placed in clinical reports to provide a bridge between 'results' and 'discussion' -it is this section that would contain the Bayesian perspective. PMID- 8248660 TI - The application of large Gaussian mixed models to the analysis of 24 hour ambulatory blood pressure monitoring data in clinical trials. AB - We propose the use of Gaussian mixed models to analyse statistically 24 hour ambulatory blood pressure data from clinical trials. We develop specific models and apply them to data from a clinical study that compares two angiotensin converting enzyme inhibitors. We investigate and discuss computing issues related to the implementation of such methods. We conclude that this methodology provides a sophisticated but practical approach to the analysis of such data. PMID- 8248661 TI - Dispersion tests and adjustments for survival and case-control studies. AB - Cox and Snell have pointed out that when the empirical variance of a measured response y is quite different from its nominal variance under an assumed probability model, standard errors that are not adjusted for the discrepancy may yield misleading inferences. This article describes a test for assessing departures from nominal dispersion in survival analysis and case-control studies. Also, a robust procedure for adjusting standard errors for these models is given. PMID- 8248662 TI - A strategy for assembling samples of adult twin pairs in the United States. AB - In this paper we develop a methodology for the identification of large numbers of U.S. adult twin pairs. Data for this study derive from the U.S. Department of Defense and the Vietnam Era Twin (VET) Registry. The Department of Defense identified potential male twins (n = 10,002) using a computerized record linkage algorithm based on the same last name, same date of birth, and the same first five digits of the Social Security number. Twinship was confirmed by comparison with the Vietnam Era Twin Registry. We developed a logistic regression model that predicts the probability that a paired record identifies twins based on the absolute difference in the last four digits in the Social Security number, the age of issuance of the Social Security number, and the frequency of occurrence of the last name. We used the estimated coefficients derived from this regression model to assign predicted probabilities of being a twin to each matched record. There is a close correspondence between the observed and expected number of twins when evaluated across deciles of predicted probabilities of being a twin; the value of the Harrell's c index (c = 0.68 +/- 0.0004) indicates the overall predictive accuracy of the regression equation. The results from this study demonstrate the feasibility of identifying adult male-male twin pairs from any large computerized database that contains name, date of birth and Social Security number. However, the selection criteria used in the creation of the computer database must be clearly specified to avoid constructing a biased sample of twins. PMID- 8248663 TI - Conditional independence models for epidemiological studies with covariate measurement error. AB - We construct a unifying representation of the structure of measurement error problems with particular reference to situations commonly encountered in epidemiological studies, and outline how estimation of the parameters of interest can be carried out in a Bayesian framework using Gibbs sampling. We show how this approach can be implemented for designs involving continuous measurement errors assessed through a validation substudy, and discuss our results on simulated data. PMID- 8248664 TI - A comparison of the generalized estimating equation approach with the maximum likelihood approach for repeated measurements. AB - Liang and Zeger proposed an extension of generalized linear models to the analysis of longitudinal data. Their approach is closely related to quasi likelihood methods and can handle both normal and non-normal outcome variables such as Poisson or binary outcomes. Their approach, however, has been applied mainly to non-normal outcome variables. This is probably due to the fact that there is a large class of multivariate linear models available for normal outcomes such as growth models and random-effects models. Furthermore, there are many iterative algorithms that yield maximum likelihood estimators (MLEs) of the model parameters. The multivariate linear model approach, based on maximum likelihood (ML) estimation, specifies the joint multivariate normal distribution of outcome variables while the approach of Liang and Zeger, based on the quasi likelihood, specifies only the marginal distributions. In this paper, I compare the approach of Liang and Zeger and the ML approach for the multivariate normal outcomes. I show that the generalized estimating equation (GEE) reduces to the score equation only when the data do not have missing observations and the correlation is unstructured. In more general cases, however, the GEE estimation yields consistent estimators that may differ from the MLEs. That is, the GEE does not always reduce to the score equation even when the outcome variables are multivariate normal. I compare the small sample properties of the GEE estimators and the MLEs by means of a Monte Carlo simulation study. PMID- 8248665 TI - Risk ratio and rate ratio estimation in case-cohort designs: hypertension and cardiovascular mortality. AB - Multivariate analysis in case-base designs depends on approximate methods. In the present study, new pseudo-likelihood methods are developed for this design. With these methods, the case-cohort risk ratio and rate ratio as well as their standard errors are easily estimated using logistic regression and Poisson regression, respectively. This is illustrated by the association between hypertension and cardiovascular mortality in a cohort, estimated by case-cohort analysis, using samples of several sizes. The estimates are compared with those obtaining in full-cohort and nested case-control designs. The results indicate that these methods, which require nothing but widely available computer software, are valid. The case-cohort design, therefore, is a good, sometimes even advantageous alternative to the nested case-control design, in studying a disease that is not very rare. Application of the risk ratio method to the full cohort, using a 'sample' of 100 per cent follows logically; whenever the true risk ratio is desired instead of the odds ratio, a multivariate model for its estimation is therefore available. PMID- 8248666 TI - The effects of professional group supervision of nurses--utilizing the nurse self description form. AB - All the registered nurses in two orthopaedic wards participated in a four-month period of professional group supervision. The main focus of the supervision was professional identity. The effect was measured by the Nurse Self Description Form, a 21-item paper-and-pencil form for nurses to judge themselves in comparison to other nurses. The nurses answered the form before and after the period of supervision. They increased their total scores significantly. Some single items were influenced more than others. The results argue in favour of group supervision as a useful mean of strengthening the professional identity of nurses. PMID- 8248667 TI - Caregiver-infant interaction--comforting post-operative neonates. AB - Analysis of 40 h of videotaped caregiving to four male infants was conducted using ethology. All infants were full term and had major chest surgery 12 h previously. All incidents of caregiver-infant interaction were examined, and the verbal and tactile comforting behavior of the caregiver was coded for duration, context, infant cue and infant state prior and following the interaction. A total of 191 comforting incidents occurred during the study. Stroking was the most common form of touch used. Most of the comforting (60%) was given in the context of caregiving; sleeping infants were touched often (38%); 25.3% of the touches were given to acutely distressed infants, and wide awake infants relatively little. Most of the infants showed no change in infant state after the relatively brief comforting incidents, but in 15.7% of the incidents the infants 'settled' after comforting. The authors conclude that this is a relatively strong effect considering the short duration of the comforting actions. PMID- 8248668 TI - Quality of care from a patient perspective--a grounded theory study. AB - The aim of the present study was to develop a theoretical understanding of quality of care from a patient perspective, using a grounded theory approach. Thirty-five interviews were conducted with a sample of 20 adult hospitalized patients (mean age: 60 years) in a clinic for infectious diseases. Data were analysed according to the constant comparative method. A model was formulated according to which quality of care can be understood in the light of two conditions, the resource structure of the care organization and the patient's preferences. The resource structure of the care organization consists of person related and physical- and administrative environmental qualities. The patient's preferences have a rational and a human aspect. Within this framework, patients' perceptions of quality of care may be considered from four dimensions: the medical-technical competence of the caregivers; the physical-technical conditions of the care organization; the degree of identity-orientation in the attitudes and actions of the caregivers and the socio-cultural atmosphere of the care organization. The model is discussed in relation to existing theories in the field. PMID- 8248669 TI - What is the meaning of caring? PMID- 8248670 TI - Adapting incontinent patients incontinence aids to their leakage volumes. AB - The importance of adapting incontinence aids according to the patients leakage volume and comfort was investigated in 28 urinary incontinent 85-year-old men and women who were living at home. The patients incontinence was quantified by a 48 hour pad test and was graded as slight (max. leakage per pad < 5 g; total leakage/48 h < 30 g), moderate (max. leakage per pad 5-15 g; total leakage/48 h 30-70 g) or severe (max. leakage per pad > 15 g; total leakage/48 h > 70 g). Four women had slight urinary incontinence, eight were moderately incontinent and nine were severely incontinent. The corresponding figures for the seven men were as follows: slight, one; moderate, one; severe, five. After the primary assessment, incontinence aids were prescribed based on the measured leakage volumes. The correctly selected and adapted incontinence aid brought the patient better comfort and security. The importance of careful information and instructions how to apply the pad is emphasised. There is also a need to see the patient again at intervals as leakage volumes may vary. Thus, smaller packages of pads should be supplied as the choice of pad may need to be modified. The latter is also important from a financial point of view as the expense of otherwise wasted pads would unnecessarily increase costs. PMID- 8248671 TI - Mental wellbeing of nurse educators at work. AB - The aim of the study was to assess the mental wellbeing of nurse educators at their work by means of self-assessment. Questionnaires were sent to 100 randomly selected trained nurse educators. Data were collected from 83 respondents. The nurse educators evaluated their mental wellbeing as being rather good. There were, however, some contradictions in the answers to questions covering identical work areas but having somewhat different formulation. The disadvantages of being a nurse educator, as listed by the respondents, included: too much work, and especially too many administrative duties, communication breakdowns with the directors of the nursing college, and underestimation by others of the value of teaching as such. As characteristics of the job promoting a sense of wellbeing, the nurse educators mentioned their interaction and relationships with the students. In addition, the demands and challenges of the work were found very interesting and rewarding. PMID- 8248672 TI - Personality development during nursing education--a Swedish study. AB - The aim of the study was to explore personality development during nursing education. The sample consisted of 122 Swedish nursing students. Ninety-three per cent of the students were female and the mean age at the beginning of the nursing education (which lasts for two years) was 27.6 years. Personality inventories were administered at the beginning and end of the education. The post-education assessment rendered several statistically significant differences of means. Compared with the pre-education assessment, the nursing students reported a more positive profession-oriented self-perception at the end of the education, a stronger ideal, real, and mirror identity, a reduced trait anxiety, stronger needs for achievement, exhibition, autonomy, dominance, and nurturance, and a weaker abasement need. The practical significance of the results was discussed. PMID- 8248673 TI - Functional status of stroke patients in long-term care--a basis for development of rehabilitation and care. AB - Thirty persons who had remained in long-term care for one year or more post stroke were assessed with respect to motor function, cognitive, perceptual, and communication ability and ADL capacity. Before the stroke they had lived an independent life, although 40% suffered from cardiovascular disease. At the follow-up, all patients had severe impairments due to the brain damage. None could walk or stand independently, ten could not call for help or attract attention in any way, and 13 could not take part in a conversation. Eleven patients had severe pain in spite of treatment with analgesics. Pain treatment and training methods that can reduce dependence and improve the quality of life for patients like these need to be developed. More appropriate assessment instruments for this patient group have to be constructed to make evaluation of training and care possible. PMID- 8248674 TI - Complications after surgery as expressed by patients and health care--the Project Peri-Operative Risk (PROPER). AB - In the present study, which is a part of a clinical-epidemiological study of complications, (the Project Peri-Operative Risk, PROPER), 510 patients, admitted for elective surgery, participated. During their time in hospital, the incidence of complications was registered by nurses. Three months after surgery, the patients were asked in a questionnaire if they had, during their time in hospital or during their convalescence at home, suffered from postoperative complications. Statements by patients and health care professionals regarding the nature and frequency of complications have been compared. Our results confirm the findings that a prospective method in registering complications results in the detection of a higher incidence than if a retrospective method is used. The analysis also reveals that patients and professionals report different types of complications. They also describe complications from different perspectives and present identical complications in different terms. PMID- 8248675 TI - [Acidosis of renal origin in childhood]. PMID- 8248676 TI - [What can one expect from the correction of nose deformities as sequelae of a unilateral cleft]. PMID- 8248677 TI - [Treatment of ovarian cancer. Literature review]. PMID- 8248678 TI - [Animal experimentation, vivisection and various other procedures]. PMID- 8248679 TI - [Therapeutic note. The treatment of oat cell bronchial cancer (1993)]. PMID- 8248680 TI - Practical use of ratings of perceived exertion in a clinical setting. PMID- 8248681 TI - Cardiovascular benefits of improved exercise capacity. AB - The evidence suggesting that regular exercise can help to prevent coronary artery disease is overwhelming. While some studies have suggested that exercise will not provide health benefits, our inactive population needs to pay heed to the substantial data presented by the many international health organisations suggesting the opposite. The American Heart Association Medical/Scientific Statement on Exercise emphasised the large role regular aerobic physical activity plays in the prevention of cardiovascular disease. Several human studies have also demonstrated the positive effects of long term exercise on the human heart. For example, it has been shown that a consistent exercise programme can lessen the impact of atherosclerotic plaques through increasing coronary artery diameter. Echocardiography studies on a training group of competitive swimmers have shown that exercise training can induce rapid changes in left ventricular dimensions and mass, which can ultimately lead to an increased stroke volume and increased maximal oxygen consumption. Studies on sedentary individuals have also demonstrated an increase in maximal oxygen uptake with a regular endurance exercise programme. In addition to these health benefits, habitual dynamic exercise can also decrease the likelihood of a cardiac event. Others have demonstrated a 50% lower incidence of coronary events in those individuals maintaining rigorous activity 2 days a week. With the preponderance of evidence revealing the health benefits of habitual exercise, it is striking to learn that more than 50% of the US population exercises for less than 20 minutes, 3 days a week. The widespread nature of this sedentary lifestyle makes inactivity an attributable fraction of the total risk factors associated with cardiac disease. The amount of exercise needed to reduce the risk of coronary artery disease is a minimum aerobic workout of 30 minutes, 4 to 5 times a week, such as a vigorous walk. Comprehensive programmes promoting exercise training should be implemented at a level appropriate to an individual's capacity and need. PMID- 8248682 TI - The critical power concept. A review. AB - The basis of the critical power concept is that there is a hyperbolic relationship between power output and the time that the power output can be sustained. The relationship can be described based on the results of a series of 3 to 7 or more timed all-out predicting trials. Theoretically, the power asymptote of the relationship, CP (critical power), can be sustained without fatigue; in fact, exhaustion occurs after about 30 to 60 minutes of exercise at CP. Nevertheless, CP is related to the fatigue threshold, the ventilatory and lactate thresholds, and maximum oxygen uptake (VO2max), and it provides a measure of aerobic fitness. The second parameter of the relationship, AWC (anaerobic work capacity), is related to work performed in a 30-second Wingate test, work in intermittent high-intensity exercise, and oxygen deficit, and it provides a measure of anaerobic capacity. The accuracy of the parameter estimates may be enhanced by careful selection of the power outputs for the predicting trials and by performing a greater number of trials. These parameters provide fitness measures which are mode-specific, combine energy production and mechanical efficiency in 1 variable, and do not require the use of expensive equipment or invasive procedures. However, the attractiveness of the critical power concept diminishes if too many predicting trials are required for generation of parameter estimates with a reasonable degree of accuracy. PMID- 8248683 TI - Effects of fasting on endurance exercise. AB - Early studies agree that fasting is detrimental to overall physical performance and to endurance performance in humans; however, a study in rats reported an ergogenic effect where time to exhaustion was increased by a glycogen-sparing effect of elevated free fatty acids in blood resulting from a 24-hour fast. Later studies on humans have also found a detrimental effect of fasting on exercise endurance, with the exception of 1 study which found no difference. The discrepancy between humans and rats could not be explained by level of glycogen sparing, mode of exercise, duration of the fast, physiological differences or level of training. The intensity of exercise, and a potential placebo effect of fasting, are possible reasons for the conflicting results. Despite reduced endurance performance, fasted humans are able to exercise and maintain blood glucose homeostasis; the specific cause of an earlier onset of fatigue during a single bout of exercise in the fasted state remains unclear. PMID- 8248685 TI - Ischial apophysis injuries in athletes. AB - The diagnosis and treatment of ischial tuberosity pain in athletes include several clinical entities. These injuries include apophysitis, adult tug lesion, painful unfused apophysis, and acute and old avulsions. In some avulsion injuries a bony fragment can be seen, but they can also be x-ray-negative, especially in children. An insertional tendon rupture is also possible. Conservative treatment of apophysitis includes modification of activities and anti-inflammatory medication. Avulsions, if diagnosed early, usually heal with conservative therapy, especially if the displacement is not marked. Urgent surgical treatment is recommended in cases with total or nearly total soft-tissue hamstring muscle insertion rupture. Surgery has to be considered also in cases with dislocation of the apophysis or bony avulsion of more than 2cm. Excision of the excessive mass or poorly united fragment provides relief of pain in some cases of old avulsions. PMID- 8248686 TI - [What is your diagnosis? Fracture of C6 and C7 spinal process in Bechterew ankylosing spondylitis]. PMID- 8248687 TI - [Endothelial dysfunction and coronary heart disease. Interaction of endothelium and thrombocytes]. AB - Aggregating thrombocytes release various mediators acting on endothelium and muscle cells of coronary arteries. In coronary arteries without endothelium, platelets induce potent vasoconstriction, whereas in segments with endothelium platelets induce relaxation. This endothelium-dependent relaxation induced by platelets is caused by the endothelial relaxation factor EDRF or nitric oxide (NO). NO inhibits not only smooth muscle cells of the vessels but also platelets themselves, an effect which is potentiated by prostacyclin. The physiologic importance of platelets-mediated contraction after removal of the vascular endothelium concerns the vascular phase of hemostasis. With the appearance of functional endothelial disorders, particularly in patients with cardiovascular risk factors, disturbances of coronary circulation can arise and finally cause ischemia. The finding that regenerated endothelial cells show functional defects especially towards the action of platelets (mainly the serotonin released from platelets) is important. In this functional disorder a dysfunction of Gi-protein of the endothelial 5HT1-serotonergic receptor plays a decisive role. Similar functional endothelial disorders can be shown after ischemia and reperfusion as well as in arteriosclerosis. PMID- 8248684 TI - Metabolic adaptations to exercise in the cold. An update. AB - Metabolic adaptations to exercise in a cold environment include the liberation of heat by vigorous physical activity, shivering and various forms of nonshivering thermogenesis. During a single exposure to cold the main metabolic fuel is glycogen; however, repeated bouts of exercise in the cold also result in an increase in fat metabolism. Potential contributors to fat loss induced by exercise in the cold include: the energy cost of synthesising lean tissue; cold induced excretion of ketones; stimulation of resting metabolism; and the high energy cost of movement in a cold environment (walking over snow, the weight of heavy boots, hobbling by winter clothing, and decreased mechanical efficiency of dehydrated muscles). Biochemical explanations of fat mobilisation include increased secretion of catecholamines, increased sensitivity of peripheral catecholamine receptors and a decrease in circulating insulin levels. Such fat loss may be helpful in treating moderate obesity, although the response seems less well developed in women than in men. Metabolic changes must be taken into consideration in preparing winter athletes for competition. Glycogen depletion has a negative effect on the performance of endurance competitors, but this can be countered by a combination of diet, training and cold acclimation. PMID- 8248688 TI - [Nitrogen monoxide (NO)--the active principle of organic nitrates]. AB - Although organic nitrates have been used in the treatment of patients with angina pectoris for more than 100 years, their mechanism of action was only disclosed during the last years. In the seventies it became already clear that nitrates act via the intracellular messenger cyclic 3,5'-guanosine-monophosphate (cGMP). Later on, S-nitrosothiols were suggested as possible intermediary messengers arising during metabolism of the nitrates. In parallel with the discovery of the endothelium-derived relaxing factor (EDRF) and its biochemical identification as nitric oxide (NO), it became clear that organic nitrates act via the release of NO in the vascular wall and thus by using metabolic pathways identical to those of endogenous EDRF. The target-enzyme for nitrates or for the NO released by them, respectively, thus is the soluble guanylylcyclase. The rate of enzymic stimulation induced by a given nitrate correlates closely with the rate of measured NO production from the nitrate molecule. The highest NO production was detected with nitroglycerin, followed by the group of dinitrates and mononitrates. In the vessel wall both endothelial cells as well as vascular smooth muscle cells can transform nitrates into NO. This might explain, why the antiaggregatory effect of nitrates is more pronounced in the presence of these cells or in vivo than it is in the absence of vascular cells in vitro. In spite of some differences in metabolism, nitrates are thus closely related by their end product to the endothelium-derived relaxing factor and represent, therefore, an adequate substitution for NO missing in diseased blood vessels. PMID- 8248689 TI - [Endogenous and therapeutic nitrates in healthy and arteriosclerotic blood vessels]. AB - Nitrates belong to the most potent drugs for treatment of angina pectoris. Patients with this disease always show arteriosclerotic changes in their coronary arteries. In arteriosclerotic vessels endothelium-dependent relaxation induced by the endogenous vasodilator nitric oxide (NO) is markedly reduced. This article introduces a theory, whereby NO production is not reduced in arteriosclerotic vessels (in the arteriosclerotic aorta of the rabbit it is even enhanced) but that the activity of superoxide dismutase, which normally metabolizes free oxygen radicals (O2-) is reduced and that NO is metabolized by accumulating superoxides. It is interesting that only endogenous nitrate (NO) can be metabolized by this pathway, whereas in arteriosclerotic vessels exogenous nitro-vasodilatators remain fully effective. For the clinical use of nitrates it is of particular importance that these exhibit different potencies for epicardial coronary vessels and the myocardial microcirculation. Nitroglycerin causes extensive relaxation in isolated large coronary arteries but has only a limited effect on middle-sized vessels and almost none on arteries in the microcirculation. These differences in efficacy of nitroglycerin in the coronary micro- and macrocirculation is probably due to the inability of microvessels to transform nitroglycerin into active metabolites such as S-nitroso-L-cysteine. Comparing vascular and clinical effects of nitroglycerin (in particular dilatation of coronary vessels) and of adenosine (acting in particular on the coronary microcirculation) discloses the great importance of the aforementioned effects on epicardial coronary vessels for treatment of angina pectoris. In this context a vasodilatating effect on the coronary microcirculation might not only be disadvantageous but could even trigger angina pectoris itself. PMID- 8248690 TI - [Clinical significance of endothelial dysfunction: studies of human coronary circulation in vivo]. AB - One of the most fascinating findings concerning our knowledge on the physiology of the cardiovascular system, especially the coronary circulation, concerns the modulating capacity of the endothelial cells on the tone of smooth muscle cells. This phenomenon was described for the first time in the aorta of a rabbit in 1980 by Furchgott and Zawadzki using acetylcholine as stimulator. Later, however, it became clear that other substances that are likewise of interest in the context of coronary heart disease, like thrombin, serotonin and histamine, have the same effect on endothelial cells. These effects can also be demonstrated in the human coronary circulation during cardiac catheterization. While normal coronary segments show a vasodilatation (corresponding to the endothelial relaxation in vitro) in arteriosclerotic coronary segments, vasoconstriction is induced by acetylcholine and serotonin. A disturbance of this vasomotor process mediated by endothelial vasomotor processes may be of particular importance in instable angina pectoris and for the development of myocardial infarction. PMID- 8248691 TI - Diagnostic advances in APUDomas and other endocrine tumors: imaging and localization. AB - The different diagnostic imaging modalities available for determining the location of the various APUDomas are discussed with reference to their advantages and disadvantages. The ability to image these lesions, and to be confident in their role in the underlying pathophysiology, and clinical neuroendocrine syndrome causing symptomatology and illness, has proved to be the key to successful treatment. In many instances it is not the diagnosis that is in question, but it is the extent of disease, the location of the lesion, or whether the anatomical abnormality under study is responsible for the complicating symptoms being considered, that are the crucial questions. The ability to locate APUDomas by a variety of direct and indirect imaging or regionalizing modalities, represent a magnificent advance in the management of these tumors. It should, however, be kept in mind that no one modality or diagnostic method can uniformly be relied upon. It is evident that a multimodal, interactive medical team approach is essential to the successful overall management of patients afflicted with these fascinating tumors. PMID- 8248692 TI - [Female non-surgical sterilization: use of endo-uterine quinacrine and betamethasone pellets. Review of the subject: experimental bases--pharmacology- toxicology--efficacy]. AB - Quinacrine produces obstruction of the region of the uterine tube by its fibroblastic granulomatous action. Factors such as the Zn++ and Cu++ content of tubal tissue, as well as the use of anti-prostaglandins, potentialise this action. Two groups of patients were evaluated in this study: Group A--95 women. 180 mg of quinacrine +0.6 mg of betamethasone were inserted into the uterine cavity, on the basis of two insertions separated by a one month interval. Two pregnancies were found by the end of the 2nd year, the percentage failure rate being 2.2 and the Pearl index 0.58. Group B--129 women. 216 mg of quinacrine +1.2 mg of betamethasone were inserted into the uterine cavity, on the basis of two insertions separated by a one month interval. Two pregnancies were found by the end of the 1st year, the percentage failure rate being 0.59 and the Pearl index 0.54. Iatrogenic adverse reactions associated with quinacrine only were not seen in either of these two study groups. None of the pregnancies was ectopic. PMID- 8248693 TI - [Cervical ripening : comparison of three methods. Preliminary results of a randomized prospective study]. AB - The authors report the preliminary results of a randomised prospective study begun in April 1992 at the maternity unit of the Caen Regional Hospital group. The aim of the study was to compare the efficacy and tolerability of three methods of cervical ripening in patients at term in whom there was an indication for the induction of labour while local conditions were unfavourable (Bishop score less than 6). In the first group, patients received an intracervical instillation of prostaglandins E2 (Prepidil), in accordance with the usual method of the department. In the patients of the second group, an intracervical Foley catheter with its balloon inflated to 50 cc was inserted and left under traction for 12 hours. Management of patients in the third group consisted of an extraamniotic injection of prostaglandins E2 via a Foley catheter previously inserted into the cervix and the balloon of which was also inflated with 50 cc of sterile water and which was left in place for 12 hours. Regardless of the method, each patient could undergo 3 ripening procedures separated by 24 hour intervals, provided the Bishop score did not reach 6. These results, which concern the first 184 patients enrolled in the protocol show the superior efficacy of the catheter + Prepidil method as compared with the other two (75% success rate after ripening as compared with 50% in each of the other two groups) as well as that intracervical Prepidil is equally effective in terms of success/failure as an intracervical catheter alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248694 TI - [Rehabilitation during pregnancy and in the postpartum]. AB - Pregnancy and labour are notable etiopathogenic factors concerning urinary incontinence and pelvic positional problems. 30 to 40% of women suffer from incontinence during pregnancy and 20% in the post-partum period. While this incontinence regresses spontaneously in 60 to 80% of cases, it nevertheless reflects the existence of pelviperineal problems. Combined with the efforts of everyday life, this trauma finally explains the high incidence of women with urinary problems and pelvic positional difficulties starting from the fourth and fifth decades of life. Perineo-sphincter exercises must be started very early, during the pre-partum phase and always before the start of sports and occupational activities. It involves all the constituents of the lumbo-pelvi perineal region and in particular perineal tone, muscle power and strength and the quality of sphincter closure during effort. These efforts are themselves studied on the basis of the pressures which they produce. In total, perineal exercises, abdominal exercises and physical activity are not contradictory but, in fact, complementary, such that the management approach is global and adapted to the individual. PMID- 8248695 TI - [Thrombophlebitis of the ovarian vein. New therapeutic approach]. AB - The authors report two cases of puerperal right ovarian vein thrombophlebitis (POVT) with floating thrombus in the inferior vena cava (IVC). The originality of this report lies in the first line surgical treatment approach. POVT is recognized as presenting usually within the first week post-partum after about 0.05% of deliveries. The syndrome consists of lower abdominal or flank pain, unexplained fever and a tender abdominal mass. Abdominal or pelvic findings are often scanty. In some cases, the thrombus may extend to the inferior vena cava, leading to the risk of pulmonary embolism or low grade renal insufficiency. Diagnosis has been difficult in the past. Since acute appendicitis is the commonest differential diagnosis, laparotomy is frequent. CT scan provides a readily available, accurate, non invasive technique for the diagnosis of POVT. Criteria are: enlargement of the vein, a low density lumen within the vessel wall and a sharply defined vessel wall enhanced by contrast media. The treatment of POVT is initially medical. Antibiotics should be given to cover the commonest infecting organisms. Heparin should also be prescribed at therapeutic IV doses to be followed by oral anticoagulants for at least six weeks. Surgery is usually only recommended when the patient remains symptomatic despite proper medical management, develops clinical, scan or arteriographic evidence of pulmonary embolism, or cannot be anticoagulated. The recommended surgical technique is to clamp the anastomosis of the ovarian vein with the vena cava.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248696 TI - [Acute pancreatitis and pregnancy]. AB - Acute pancreatitis during pregnancy is a serious condition and diagnosis is often difficult. The authors report the case of a 32-year-old woman in the 32nd week of her fifth pregnancy, in which the outcome was fatal for both mother and child. The cause of pancreatitis during pregnancy has been attributed to many factors, chiefly cholelithiasis. A number of recent studies have shown the relationship existing between the role played by pregnancy in predisposing to gallbladder disease with lithiasis. Many diagnosis errors are made in this condition. Thus modern treatment methods have improved the prognosis in acute pancreatitis but, when it occurs during pregnancy, diagnostic delays often lead to a gloomy outlook. PMID- 8248697 TI - [Alcoholism: addiction or disease?]. PMID- 8248698 TI - [Protective effect of pancreatic enzyme depletion in the course of acute pancreatitis--experimental study in rats]. AB - This study was performed to assess the effects of the reduction of pancreatic enzymatic content in the evolution of acute pancreatitis (A.P.) in rats. We hypothesized that cerulein in physiologic dose reduces the enzymatic content of the pancreas and may decrease the mortality of A.P.. Four groups of rats have been studied. In the group I cerulein in a total dose of 8 ug/Kg was given, Group II was a control group, Group III received cerulein in inframaximal dose (0.4 micrograms/Kg) and Group IV was a control group with free access to diet and water. Acute pancreatitis was induced by retrograde injection of taurocholic acid into the pancreatic duct. The mortality was 60% in group I, 56% in group II, 23% in group III and 34% in group IV. These results suggest that by decreasing enzymatic content of the pancreas the severity of acute pancreatitis may be reduced. PMID- 8248699 TI - [Tropical myositis]. AB - The authors describe six cases of tropical myositis or pyomyositis treated in the Department of Internal Medicine of the University of Sao Paulo between 1985 and 1992. Staphylococcus aureus was the causative agent. It was isolated from the muscular abscess in four cases and blood cultures in two cases. The treatment with appropriate antibiotics and drainage of the abscess(es) determined satisfactory evolution without mortality or residual deformity. PMID- 8248700 TI - [Effect of quinolones on mice experimental infection by Plasmodium berghei and their possible therapeutic usefulness]. AB - The search for new antimalarial drugs is important for many reasons, specially because of the resistance of plasmodia. Some clinical and laboratory studies have recently indicated that quinolones, currently in use for treatment of bacterial infections, have antimalarial activity. So, we evaluated the possible action of ciprofloxacin, norfloxacin, ofloxacin and pefloxacin in mice experimentally infected by Plasmodium berghei, by the oral route. Taking into account parasitemia and mortality, we came to conclusion that these drugs are not effective, as judged by the methods used. PMID- 8248701 TI - [Gastric wounds: study of 85 patients]. AB - During January 1987 and December 1990 we studied 85 patients (75 males), the age varying between 6 and 47 years (mean 27.6) sustaining penetrating (81 patients) and blunt (4 patients) gastric wounds. The mortality rate was 17.6% (15 patients) with four deaths occurring during the surgical procedure as consequence of critical associated injuries. Only one death happened as consequence of gastric wound; 6 patients died during the first 24 hours of hospitalization because of irreversible shock, 9 deaths came about after the first day of hospitalization (4 sepsis, 3 respiratory failure, 2 head trauma). The morbidity rate among the 81 patients that survived after surgical treatment was 39.5% (32 patients). The main postoperative gastric complication was vomiting in 10 patients (5 presenting vagus nerve injury and 5 sustaining pancreatic and/or another hollow viscus wounds) one patient presented with gastric suture dehiscence. Among the 12 patients sustaining vagus nerve injury the presence of gastric complication was higher in the group that was not submitted to pyloroplasty (6 patients). Analysing the patients presenting gastric and pancreatic injuries we verified that morbidity rate was statistically significantly higher in this group (69.2%) than in overall morbidity (39.5%). PMID- 8248702 TI - [Simultaneous injury of 4 iliac veins by firearm: report of a case and review of the literature]. AB - Iliac vessel injuries constitute approximately 10 percent of all vascular traumatic lesions. The high mortality rate is due to the massive exsanguination and the high frequency of associated injuries. Iliac vein injury occurs either isolated or in association with the arterial lesion. Multiple injuries of iliac veins are uncommon and frequently fatal. An unusual case, in which there were injuries of the four iliac veins by gunshot is described. A review of literature of this condition and a discussion about the diagnosis, treatment and factors affecting mortality rate is presented. No other such case has been found reported. PMID- 8248703 TI - [Congenital malaria: report of a case and considerations about the subject]. AB - We report a case of Plasmodium falciparum malaria in a 21 day old child that was born from a mother that developed clinical signs of falciparum malaria just after giving birth and died. Transmission could have occurred either by the transplacental route or by maternal blood reaching the child during the childbirth. Literature review shows that malaria should be considered as a cause of a febrile disease in the newborn, particularly if there is a low birthweight and the newborn is a child of a woman coming from an endemic malaria region. PMID- 8248704 TI - Polyvalent pneumococcal polysaccharide vaccines; a review of the literature. AB - Streptococcus pneumoniae is a leading cause of morbidity and mortality, mainly in children, elders and individuals with AIDS or AIDS-related complex, being a frequent bacterial cause of pneumonia, otitis media, sinusitis, bacteremia and meningitis. Polyvalent pneumococcal capsular polysaccharide vaccines contain the 23 most common pneumococcal serotypes causative of pneumococcal infection in several countries. Public Health Service Advisory Committee on Immunization Practice of the Centers for Disease Control (CDC) does not recommend the polyvalent pneumococcal vaccine for general public. However, several investigators have recommended its employment for special population at high risk, such as for HIV infection, who can be at enhanced risk for systemic pneumococcal disease. The objective of the present literature review is to relate the importance of studied different types of polyvalent pneumococcal vaccines as well as their immunological properties in the vaccinated people. PMID- 8248706 TI - [A European measure of health: the EuroQol]. PMID- 8248705 TI - [Typology of performance of graduate course disciplines in medicine at the Medical School of the University of Sao Paulo]. AB - The results of the program for the evaluation of the curriculum of the graduation course of the Medical Faculty of the University of Sao Paulo are presented and discussed. Following disciplines received special attention: practice in a hospital for infections diseases, orthopedics and traumatology, medical imagery, propedeutics and preventive and social pediatrics. These disciplines were chosen in an aleatory way among those that correspond to the four models of the behaviour: increasing, decreasing, decreasing/increasing and increasing/decreasing. PMID- 8248707 TI - [Hypertension. A behavioral treatment]. PMID- 8248708 TI - [Nursing diagnoses. An instrument for administering care]. PMID- 8248709 TI - [What is the Sant Gregori Consortium and how does it operate?]. PMID- 8248710 TI - [Nutritional study in a center for the mentally retarded]. PMID- 8248711 TI - [Pharmacognosy. An emerging field of knowledge]. PMID- 8248712 TI - [Endocarditis and valve prostheses]. PMID- 8248714 TI - [A "catalog of health benefits". Rationalizing the health system]. PMID- 8248713 TI - [Continuing education. 83. Medical-surgical. Transient cardiac stimulation]. PMID- 8248715 TI - Development of an animal model for testing human breast implantation materials. AB - Although breast implant materials have been tested in laboratory species since the early 1950s, a standardized evaluation system does not currently exist in which human-made polymers are exposed directly to the mammary milieu of female animals. The present study evaluated such a model as the basis for future experiments on long-term tissue effects. Polyesterurethane disks, 6 mm in diameter x 3 mm thick, were inserted bilaterally beneath the axillobrachial and inguinal mammary/fat pads of 50 9-wk-old female B6D2F1 mice (4 implants each). Implant sites were examined histologically at time points 24 hr to 47 wk after surgery. An acute inflammatory reaction at the implant edges began within 24 hr, and macrophages were found lining the smooth polyurethane fiber surfaces at the periphery by day 2. Multinucleated foreign body giant cells formed by day 4, and by week 4 giant cells contained polyurethane fragments within the cytoplasm, implying degradation of the material. Implant sites showed declining subacute inflammatory responses and increasing fibrosis by week 5. By 13 wk, the polyurethane disks appeared to be integrated into the growing adipose and mammary tissues. Although not apparent on gross inspection, microscopic examination showed that polyurethane fibers moved progressively into adjacent tissues and were always associated with chronic granulomatous inflammation. Histologic findings in the present study are strikingly similar to the human response to polyurethane-coated breast implants. These results suggest the applicability of this model to appropriately test mammaplasty materials in mammary tissues. PMID- 8248716 TI - Pituitary carcinoma of pars distalis as a common neoplasm in Fischer-344 rats. AB - The incidence of invasive spontaneous pituitary carcinomas in Fischer-344 rats were studied. The pituitaries were examined histopathologically together with surrounding tissues including sphenoid bone. The incidence of carcinoma was higher than that previously reported in Fischer-344 rats. All the carcinomas observed in this study showed evidence of local invasion either ventrally into bone and bone marrow or laterally into peripheral nerves and blood vessels. No dorsal infiltration into the brain was recognized. Pituitary carcinoma with local invasion is a relatively common neoplasm when examined histopathologically together with the surrounding tissues. PMID- 8248717 TI - c-N-ras is activated infrequently in canine malignant lymphoma. AB - Activated c-N-ras alleles have been detected in human lymphoma specimens. The aim of the present study was to determine the frequency of c-N-ras mutational activation in canine malignant lymphoma. DNA was isolated from 28 canine malignant lymphoma specimens collected from 28 separate dogs and examined for c-N ras mutations by polymerase chain reaction amplification and direct sequencing. The tumors were naturally occurring and derived from 20 dogs with known exposures to the phenoxy herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and from 8 dogs with no known exposure to the herbicide. An oncogenically activating mutation was found in 1 dog without known 2,4-D exposure. The mutation was a 13th codon, second position transition that would result in a glycine-to-aspartate amino acid substitution. The results of this study demonstrate that, similar to the human, c N-ras mutations are uncommon in dogs with malignant lymphoma and that there is no association between 2,4-D exposure and activation of c-N-ras in the dog. PMID- 8248718 TI - Testicular degeneration and spermatid retention in young male rats. AB - The incidence of spontaneous testicular atrophy and its morphological changes in relation to stage-specific spermatogenesis were investigated in young Crl:CD/BR male rats at 10-12 wk of age used as controls for toxicity screening during 1983 1990. The incidence of testicular degeneration was 2.5% (5/197) in control rats used for oral toxicity studies and 9.4% (31/327) in rats used for inhalation studies. The epididymal tubules of rats with testicular degeneration had exfoliated germ cells and low sperm density. The high incidence of testicular degeneration observed in the control rats used in inhalation studies may be related to the stress associated with immobilization in the restrainer during nose-only exposure conditions. The severity of testicular degeneration in the inhalation studies was mostly minimal. In these minimally affected testes, mature spermatids (step 19) were retained within normal-appearing germinal epithelium at spermatogenic stages IX-XIV. Also, eosinophilic globular bodies (EGBs) were formed with elongated or mature spermatids throughout all spermatogenic stages, but the general architecture of germinal epithelium was normal in appearance. By electron microscopy, EGBs were sequestered necrotic spermatids, and the germ cell degeneration was associated with cytoplasmic vacuolation of Sertoli cells. In moderate testicular degeneration, markedly decreased maturing spermatids (steps 15-19) and a slight depletion of round spermatids were observed in stages I-VIII. In severe testicular degeneration, seminiferous tubules were lined with 1-2 layers of round spermatids and spermatocytes with giant cell formation. The round spermatids served as a marker to identify spermatogenic stages (I-VIII) of the atrophic tubules. Also, in severe testicular degeneration, tubules in spermatogenic stages X-XIV had no elongated spermatids, and spermatocytes were exfoliated with occasional giant cell formation. Many seminiferous tubules were lined with only 1-2 layers of spermatocytes, and specific germ cell markers were not present. PMID- 8248719 TI - Tumor induction by concurrent oral administration of ethylenethiourea and sodium nitrite in mice. AB - Carcinogenic potential of ethylenethiourea (ETU) in combination with sodium nitrite was investigated in ICR mice of both sexes. Groups of 30 males and 30 females each were given 10 weekly oral administrations of ETU and sodium nitrite with the following combinations of dosing (ETU vs sodium nitrite, mg/kg/wk): 0 vs 0, 100 vs 0, 0 vs 70, 25 vs 17.5, 50 vs 35, and 100 vs 70. Thereafter, the animals were allowed to live without treatment up to 18 mo after the first administration. Concurrent administration of ETU and sodium nitrite caused earlier development of tumors and/or dose-dependent increases in the incidences of tumors in the lymphatic tissue, lung, forestomach, Harderian gland, and uterus, whereas treatment with either ETU or sodium nitrite failed to show carcinogenic activity. In addition, carcinomas in the forestomach and uterine horn were limited to mice receiving concurrent administrations of ETU and sodium nitrite. These results indicate that ETU is most probably converted in vivo into N-nitroso ETU and that the N-nitroso ETU has a greater carcinogenic potential in mice than ETU alone. PMID- 8248720 TI - Morphologic investigations of the guinea pig model of iron overload. AB - We have developed a guinea pig model of iron overload toxicity. Animals were administered intraperitoneal iron dextran 3 times a week to achieve total body iron load of 0.25, 0.5, 1.0, 1.5, and 2.0 g Fe/kg body weight in less than 30 days. Quantitation of tissue iron levels with atomic absorption indicated increased iron deposition in liver and heart of the iron-loaded guinea pigs (p < 0.001). Additionally, the iron-loaded pigs demonstrated decreased nuclear magnetic resonance spectroscopy T1 relaxation times in both liver and heart (p < 0.001). Serum iron, total body iron capacity, and transferrin saturation values were also determined in guinea pigs treated with 0.25, 0.5, and 1.0 g Fe/kg body weight. Serum iron and total iron-binding capacity were significantly increased at 0.5 and 1.0 g Fe/kg; transferrin saturation was elevated at 0.25 and 1.0 g Fe/kg. kg. Histologic examination of liver, heart, and bone marrow as well as ultrastructural studies on liver and heart confirmed increased iron deposition in treated animals. At the low iron dose level of 0.5 g Fe/kg, liver iron particles were primarily confined to Kupffer cells with minimal hepatocellular localization. Increased hepatocellular iron deposition was observed with larger doses of loaded iron. Myocardial iron was most prominent in interstitial cells of the epicardium, endocardium, myocardium, and coronary adipose tissue. Ultrastructurally, the presence of iron particles in perinuclear, membrane-bound structures (consistent with lysosomes) was confirmed using x-ray microanalysis. These morphological studies suggest that in this animal model siderosis of hepatic mononuclear phagocyte and myocardial interstitial cells may be the initial lesions leading to further biochemical and functional abnormalities. Correlation between tissue iron measurements and both light and electron microscopic changes, presented in this report, serve to introduce the iron-loaded guinea pig as a model for the study of iron-induced tissue damage. PMID- 8248721 TI - Iron-induced myocardial and hepatic lysosomal abnormalities in the guinea pig. AB - The guinea pig model of iron overload, described in the preceding article, was used to investigate the mechanism of excess iron toxicity in hepatic and cardiac tissues. Effects of iron overload on both lysosomal membrane fragility and membrane peroxidation were studied. The free activity of selected myocardial and hepatic lysosomal enzymes, in addition to serum activity, was measured in guinea pigs treated with iron dextran (0.25, 0.5, 1.0, and 2.0 g Fe/kg body weight); controls received dextran. Levels of malondialdehyde were also determined in whole homogenates of heart and liver in animals loaded with 0.5 and 1.5 g Fe/kg of iron dextran. Results indicated that the free activity of hepatic glucosaminidase (p < 0.05) and beta-glucuronidase (p < 0.05) were significantly elevated at all levels of iron loading; hepatic acid phosphatase was increased at all but the lowest iron dose. Similarly, increased serum glucosaminidase activity was observed (p < 0.01) at all dose levels. When compared to pooled controls, the free activity of myocardial glucosaminidase was also elevated (p < 0.05) at all levels of loading. However, myocardial acid phosphatase was increased only at the highest iron dose (p < 0.01). Increased malondialdehyde was measured at the high iron dose (1.5 g Fe/kg) in whole homogenates of both heart and liver (p < 0.01). We conclude that iron loading in this model profoundly alters the stability of hepatic and myocardial lysosomal membranes; furthermore, changes in serum glucosaminidase activity may be reflective of modified tissue lysosomal properties. Elevated levels of malondialdehyde in whole homogenates suggest that iron-mediated lipid peroxidation may be responsible in part for enhanced lysosomal membrane fragility. PMID- 8248722 TI - Time course of hepatic injury and recovery following coadministration of carbon tetrachloride and trichloroethylene in Fischer-344 rats. AB - Simultaneous administration of trichloroethylene (TCE), at an oral dose of 0.5 ml/kg, resulted in a marked potentiation of liver injury caused by an oral dose of carbon tetrachloride (CCl4, 0.05 ml/kg). Hepatic glutathione levels were depressed at 24 hr only in the rats given TCE and CCl4. Using serum enzyme (ALT and SDH) as indicators of hepatotoxicity, potentiation of CCl4-injury was most apparent at 24 hr. Upon histological examination of H&E stained liver sections, the differences between livers obtained from TCE and CCl4-treated rats versus CCl4-treated rats were most apparent at later time points (48 and 72 hr). At 48 hr after CCl4, livers showed a distinctive and uniform pattern of injury with regeneration features predominating over necrosis. At this time, livers from TCE and CCl4-treated rats were characterized by extensive zone 3 coagulative necrosis. Inflammatory infiltrations were less prominent. At 72 hr, morphological features of livers from TCE and CCl4 rats were similar to those from rats given CCl4 alone at 48 hr. From the results obtained, it appears that the regenerative activity of the liver may be delayed in rats simultaneously administered TCE and CCl4 as compared to rats administered only CCl4. PMID- 8248723 TI - Spontaneous erythroleukemia in a 16-wk-old female Slc:SD rat. PMID- 8248724 TI - Hypodontia in 9-year-old Norwegians related to need of orthodontic treatment. AB - The present study reports on the prevalence of hypodontia in a Norwegian population and classifies children with hypodontia according to need of orthodontic treatment. Orthopantomograms of 1953 children (960 girls and 993 boys) at the age of 9 were available for examination. Of the boys registered with hypodontia of second premolars at the age of 9, 11.3% showed late mineralization between the ages of 9 and 12. Only 2.9% of the girls showed late mineralization. The corrected prevalence of hypodontia, excluding third molars, in the girls was 7.2%, in the boys 5.8%, and in both sexes combined 6.5%, the difference between sexes not being statistically significant. Of the children with hypodontia, 86.6% lacked only one or two permanent teeth. The most frequently missing teeth were the mandibular second premolars, the maxillary second premolars, and the maxillary lateral incisors, in that order. Classification of children with hypodontia according to need of orthodontic treatment showed that about two thirds had hypodontia only of single posterior teeth with a moderate need of treatment. About one-third had hypodontia involving anterior teeth, and only 3.1% had hypodontia of two or more teeth in the same quadrant with a great need of treatment. PMID- 8248725 TI - Dentin formation in miniature pigs with special reference to indomethacin and orthodontic treatment. AB - The rate of dentin mineralization and the influence of indomethacin on the dentin mineralization rate during orthodontic treatment was determined in miniature pigs by intravital labeling with tetracycline. The results demonstrated that the dentin mineralization rate in the control animals was 3.8 microns/day, a rate corresponding to that of human teeth. Both indomethacin and orthodontics had an effect on the dentin mineralization rate, indomethacin reducing and orthodontic forces increasing it. In combination, the two factors neutralized each other. PMID- 8248726 TI - Exaggerated abrasion/erosion of human dental enamel surfaces: a case report. AB - An atypical, rapidly proceeding abrasion/erosion of the labial enamel surfaces of the maxillary and mandibular incisors and canines in a 27-yr-old man is reported. Ultrastructural examination of a replica of the teeth showed a practically structureless enamel surface both at the initial examination and after 12 months. However, at the end of the period, minor areas of dentin tubules became visible, indicating that a substantial loss of the tooth substance had taken place. The patient's occupation involved daily environmental contact with proteolytic enzymes. In vitro study of enamel exposed to one of the actual proteolytic enzymes showed dissolution of enamel substance, and it cannot be excluded that enzymatic decomposition of the organic enamel matrix is a contributing cause of the observed exaggerated loss of tooth substance. PMID- 8248727 TI - Effectiveness of caries removal by the partial tunnel preparation method. AB - The tunnel preparation method is designed to remove approximal caries through a channel from the occlusal surface while preserving the marginal ridge. This method entails reduced access to the caries lesion and thereby uncertainty as to the complete removal of caries. The purpose of the present investigation was to study the effectiveness of caries removal in 60 extracted premolars and molars by the partial tunnel preparation method. The glass polyalkenoat (ionomer) filling and the distance to the pulp were also examined. Examination of the sectioned teeth showed residual caries in the axial wall of two teeth and in dentin close to the enamel lesion in 10 teeth. Very few porosities were found within the glass polyalkenoat material and at the interface between the filling and the cavity walls. PMID- 8248728 TI - Salivary fluoride clearance after a single intake of fluoride tablets and chewing gums in children, adults, and dry mouth patients. AB - The aim of the present investigation was to compare the clearance pattern in saliva and the salivary stimulating effect of a new fluoride (F) chewing gum (Fluorette) with three other F products used in Scandinavia for many years for caries prevention. Concentration of F was determined in whole saliva in three groups of subjects: 1) children, 10-12 yr of age (n = 20), 2) adults (n = 20), and 3) dry mouth patients (n = 15), after a single intake of the two tablets, Dentan and Fludent, and the two chewing gums, Fluomin and Fluorette, all containing 0.25 mg F as NaF. Sucking was allowed until the tablets had been completely dissolved in the mouth. The chewing gums were used for 15 min. Saliva samples were collected from subjects expectorating 0.3-0.5 ml at nine different time intervals up to 45 min after the intake. There were some significant differences in the maximum F concentration, the area under the salivary fluoride concentration curve (AUC) when plotted against time, and the salivary stimulating effect among the four products, but as a whole they were small and probably of minor clinical importance. Among the various groups, the dry mouth patients showed the highest salivary F concentration. Thus, the main conclusion from this study is that the F tablets and chewing gums studied, including the new product Fluorette, had approximately the same clearance pattern in saliva and the same salivary stimulating effect. However, there were great variations among the different subjects. PMID- 8248729 TI - Tissue plasminogen activator (t-PA) and placental plasminogen activator inhibitor (PAI-2) in gingival fluid from 8-9-year-old children. AB - High concentrations of tissue plasminogen activator (t-PA) and placental type plasminogen activator inhibitor (PAI-2) have previously been found in gingival crevicular fluid (GCF) of adults. In the present study, the levels were examined in 16 children aged 8-9 yr. Sampling of GCF was performed with small disks of Millipore-filter. t-PA and PAI-2 were analyzed with enzyme-linked immunosorbent assays with low method errors. The mean concentration of t-PA was slightly higher than in adults, while the mean PAI-2-concentration was slightly lower. An intraindividual study comparing healthy and inflamed sites in the children showed slightly higher concentrations in GCF from inflamed sites. No change was observed in the balance between t-PA and PAI-2. PMID- 8248730 TI - Cyclosporin-A-induced gingival overgrowth in renal transplant children. AB - Gingival overgrowth was assessed in renal transplant children, 19 boys and 13 girls, aged 2.5-18 yr, who had been on a cyclosporin-A (CsA)-based immunosuppressive regimen for at least 12 months. Data collected included number of posttransplant months, total CsA dose administered during the first 6 posttransplant months, oral dose and blood trough level of CsA on the day of examination, plaque accumulation (VPI%), gingival inflammation (GBI%), loss of attachment, and gingival overgrowth. Thirteen percent of the children exhibited gingival overgrowth characterized by one or more units with increased sulcus probing depth (> or = 4 mm), i.e. pseudopockets. The total amount of CsA administered during the first 6 posttransplant months was significantly higher in the children with gingival overgrowth than in those without. The study indicates that the development of CsA-induced gingival overgrowth is positively related to the total dose of the drug administered to the children during the first 6 posttransplant months. PMID- 8248731 TI - Doxycycline prevents root resorption and alveolar bone loss in rats after periodontal surgery. AB - The effect of systemic doxycycline administration on frequency of root resorption cavities and extent of bone loss following periodontal surgery was studied in albino rats. Thirty Wistar rats with healthy gingiva were divided into four groups. Six untreated animals (Group 1) served as controls. Gingivectomy (Group 2, six rats), mucoperiosteal flap operation (Group 3, eight rats), and mucoperiosteal flap operation with doxycycline added to the drinking water (Group 4, 10 rats) were performed on the palatal aspect of maxillary left molars. After a healing period of 3 wk, the rats were killed by an overdose of sodium pentothal, and specimens of first molars were prepared for light microscopy. Root resorption was absent in normal and gingivectomized animals (Groups 1 and 2). Resorption cavities occurred in all flap-operated animals (Group 3) and in one of 10 doxycycline-treated animals (Group 4). The distance from cementoenamel junction to alveolar bone crest remained unchanged in gingivectomized (Group 2) and doxycycline-treated animals (Group 4), as compared with control specimens. In flap-operated animals (Group 3), the crestal bone level was located more apically than in doxycycline-treated rats (Group 4). This study has shown that root resorption and bone loss were associated with flap operations involving exposure of periodontal ligament and bone. Systemic doxycycline prevented both root resorption and bone loss. PMID- 8248732 TI - In vitro antimicrobial susceptibility of different serotypes of Actinobacillus actinomycetemcomitans. AB - In vitro susceptibility of Actinobacillus actinomycetemcomitans (A.a.) serotypes to selected antimicrobial agents was investigated by the agar dilution method on supplemented Mueller-Hinton test medium. Eighty-three A.a. strains, 80 recent isolates from 40 periodontally healthy or diseased subjects, and three type strains were included in the study. Serotype a represented 20, serotype b 32, serotype c 17, and serotype e 7 and nontypable 4 of the tested strains. The most effective drugs against all A.a. serotypes in vitro were cefaclor, cefuroxime, tetracycline hydrochloride, doxycycline, trimethoprim-sulfamethoxazole (cotrimoxazole), and ciprofloxacin, which inhibited 100% of the strains at 4.0 micrograms/ml, 4.0 micrograms/ml, 1.0 microgram/ml, 2.0 micrograms/ml, 0.06 microgram/ml, and 0.015 microgram/ml, respectively. Serotypes a and e were more susceptible to cefaclor and cefuroxime than were serotypes b and c; 100% of the first two groups were inhibited at 2.0 micrograms/ml and 1.0 microgram/ml. Ampicillin inhibited 92% of the tested strains at 1.0 microgram/ml. Serotype b was always susceptible to ampicillin. Metronidazole exhibited the best activity against serotype a strains. The lowest minimal inhibitory concentration values for benzylpenicillin, ampicillin, erythromycin, doxycycline, and metronidazole were encountered among serotype b isolates. The results of the present study indicate minor differences in the in vitro antimicrobial susceptibility patterns of different A.a. serotypes, except to metronidazole. Also, the new oral cephalosporins and cotrimoxazole, rare antimicrobial agents in periodontology, showed promising efficacy against all A.a. strains. PMID- 8248733 TI - Eubacterium yurii subspecies margaretiae is resistant to nonopsonic phagocytic ingestion. AB - We have previously shown that strains of Eubacterium yurii are hydrophobic, as compared with human polymorphonuclear leukocytes (PMNs), possibly because of a crystalline surface layer (S-layer) covering the cell envelope of this potential endo-perio pathogen. The aim of the present study was to investigate the phagocytic ingestion by PMNs of the three E. yurii subspecies, with special attention to bacterial surface structures and hydrophobicity. Type strains of subspp. margaretiae, yurii, and schtitka, together with three clinical isolates from necrotic root canals, were studied. All strains were hydrophobic when tested by a two-phase partition method. E. yurii subspp. margaretiae strains ATCC43715T, ES4C, and ES14B-8E were resistant to PMN ingestion in the absence of opsonins, whereas strains of the two other subspecies were readily ingested. The presence of a resistant strain (subsp. margaretiae ATCC43715T) did not inhibit the ingestion of a sensitive strain (subsp. schtitka ATCC43716T). Ingestion of E. yurii subsp. margaretiae strains required opsonization by normal human serum or specific antibodies. Electron microscopy revealed an S-layer in all strains and fimbria-like structures in the subspp. margaretiae and yurii strains. The antiserum prepared against the S-protein of E. yurii subsp. margaretiae ATCC43715T showed only slight cross-reactivity with other E. yurii strains and indicated the presence of strain-specific rather than species- or subspecies specific antigens in the S-protein of E. yurii subsp. margaretiae ATCC43715T. The results suggest that the mere presence of the S-layer or fimbria-like structures cannot explain the susceptibility to ingestion by the PMNs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248734 TI - Varicella-zoster virus DNA in recurrent aphthous ulcers. AB - The present study was undertaken to examine for varicella-zoster virus (VZV) constituents in recurrent aphthous ulcers (RAU). Smears and biopsies of ulcers 0 3 days old were obtained from 20 otherwise healthy minor/major RAU patients and from two patients with Behcet's syndrome. VZV could not be cultivated from any of the biopsies, all smears were negative for VZV antigen, and in situ hybridization on the five samples tested was negative for VZV nucleic acid. DNA extracted from biopsies, however, yielded positive polymerase chain reaction (PCR) products in all cases with one primer set of the protein kinase 66 (PK66). Ten control biopsies from non-RAU subjects yielded no PCR products apart from the sample from AP, who obtained the biopsies, and from HOM, who performed the PCR, suggesting that the DNA had been passed on from the patients. Other parts of the PK66 gene and of thymidine kinase 36 yielded no products. Although the consistent presence of VZV DNA in RAU might suggest a pathogenetic implication, the potential presence and possible pathogenetic importance of other herpesviruses cannot be excluded. PMID- 8248735 TI - Mandibular elevator muscles: physiology, action, and effect of dental occlusion. AB - In spite of differences in embryologic origin, central nervous organization, and muscle fiber distribution, the physiology and action of mandibular elevator muscles are comparable to those of skeletal muscles of the limbs, back, and shoulder. They also share the same age-, sex-, and activity-related variations of muscular strength. With respect to pathogenesis, the type of muscular performance associated with the development of fatigue, discomfort, and pain in mandibular elevators seems to be influenced by the dental occlusion. Clinical research comparing the extent of occlusal contact in patients and controls as well as epidemiologic studies have shown reduced occlusal support to be a risk factor in the development of craniomandibular disorders. In healthy subjects with full natural dentition, occlusal support in the intercuspal position generally amounts to 12-14 pairs of contacting teeth, with predominance of contact on first and second molars. The extent of occlusal contact clearly affects electric muscle activity, bite force, jaw movements, and masticatory efficiency. Neurophysiologic evidence of receptor activity and reflex interaction with the basic motor programs of craniomandibular muscles tends to indicate that the peripheral occlusal control of the elevator muscles is provided by feedback from periodontal pressoreceptors. With stable intercuspal support, especially from posterior teeth, elevator muscles are activated strongly during biting and chewing with a high degree of force and masticatory efficiency, and with relatively short contractions, allowing for pauses. These variables of muscle contraction seem, in general, to strengthen the muscles and prevent discomfort. Therefore, occlusal stability keeps the muscles fit, and enables the masticatory system to meet its functional demands. PMID- 8248736 TI - Mandibular condyle lesions related to age at onset and subtypes of juvenile rheumatoid arthritis in 15-year-old children. AB - The severity of lesions in the temporomandibular joint (TMJ) area and their association with age at onset, the various forms of juvenile rheumatoid arthritis (JRA), and certain serologic tests for rheumatoid factor (RF), antinuclear antibodies (ANA), and human leukocyte antigen (HLA)-B27 were determined in 15-yr old children. The series comprised 121 JRA children, 78 girls and 43 boys, in whom an analysis had recently been made of the relation of TMJ lesions to jaw movement and occlusal status. The mean age at onset was 7.3 yr, the girls contracting JRA earlier than the boys. Also, the girls with lesions in the TMJ area were significantly younger than those with no lesions. TMJ abnormalities were found in 50% of cases with a pauciarticular or systemic onset, but in 72% of those representing the polyarticular subtype. Flattened condyles and grave lesions were equally represented in all subgroups and in both sexes. A crossover from onset type to present diagnosis was found in 30% of the cases, mostly from pauciarticular to polyarthritis, which also increased the risk of TMJ lesions from 50 to 60%. RF, ANA, or HLA-B27 alone did not seem to be associated with a risk of TMJ abnormalities. Maximal opening capacity is more restricted in patients with early onset or a polyarticular mode of JRA. Since the TMJ is affected in more than half of JRA children, regular measurements of maximal movements of the mandible or roentgenologic examinations of the TMJ are essential for their optimal treatment. PMID- 8248737 TI - Reflectometry: a new method for quantitative determinations of intraoral film formation. AB - A simple optical method--reflectometry--is described to determine the thickness and mass of salivary films formed on solid surfaces at oral exposure. Reflectometry is based on the fact that p-polarized light is reflected with minimum intensity at an interface when the angle of incidence equals the so called pseudo-Brewster angle. Hydrophilic and hydrophobized silica surfaces were used as substrates and the reflectometer was calibrated against a null ellipsometer. A linear relation was found between the square of the ellipsometrically measured thickness of silicon oxides on silicon substrates and the reflectometrically registered intensity of the reflected light at these surfaces. Thirty volunteers participated in the study of the thickness of films formed on the test substrates at oral exposure. The test silica surfaces were positioned in the vestibular sulcus of each test subject for periods of 1, 5, 15, 30, 60, and 120 min. The results show that films formed on hydrophobic surfaces leveled-off to a thickness value of 140 A after 60 min. Films formed on hydrophilic surfaces, however, reached a plateau value of approximately 100 A after only 300 min. Reflectometry seems to be an accurate and valid, yet inexpensive and quick method for quantitative investigations and thickness measurements of early salivary pellicles in large subject groups. PMID- 8248738 TI - Alpha-hemolytic streptococcal bacteremia: a review of 203 episodes during 1980 1991. AB - This is a review of 203 episodes of alpha-hemolytic streptococcal bacteremia during a 12-year period (1980-91) in a large community teaching hospital in the USA. There were 87 male and 116 female patients, with ages ranging from newborn infants to a 96-year-old; 118 patients (58.1%) were 60 years or older. 31 patients had endocarditis, 93 had bacteremia, and 79 had polymicrobial bacteremia. Streptococcus sanguis, S. mitis, S. salivarius and S. intermedius accounted for 80% of the strains. Some strains were resistant to tetracycline, nafcillin, erythromycin or clindamycin. The overall mortality was 29.6% (60 of 203 patients). There was no significant difference in the mortality among patients with endocarditis, bacteremia, and polymicrobial bacteremia. Factors that adversely influenced the mortality rate were old age (60 years and older) and rapidly fatal or ultimately fatal underlying conditions. PMID- 8248739 TI - Hepatitis B carriers in Sweden--effects of immigration. AB - The number of hepatitis B carriers in Gothenburg, the second largest city in Sweden, has gradually increased between 1980 and 1990. Altogether 525 chronic carriers (35% females) were found, corresponding to a prevalence of 0.11%. 401 (76%) were immigrants from high or medium prevalence regions, mainly the Middle East and Southeast Asia. 102 were native Swedes (19%). In 1988-90 the immigrants constituted 85% (154/182) of all new cases. 93% (245/264) of the carriers born in 1960-89 were immigrants, whereof 53% (130/245) were HBeAg positive. The increasing number of hepatitis B carriers in Sweden places demands on the public health service to undertake measures to prevent new cases of hepatitis B, as well as to discover and treat chronic liver disease induced by hepatitis B virus. PMID- 8248740 TI - Diagnosis of cytomegalovirus in bronchoalveolar lavage by polymerase chain reaction, in comparison with virus isolation and detection of viral antigen. AB - Bronchoalveolar lavage (BAL) products from 52 immunocompromised patients with symptoms of pulmonary infection was examined for cytomegalovirus (CMV) by virus isolation, polymerase chain reaction (PCR) and detection of CMV antigen by immunofluorescence or immunoperoxidase staining after short-term incubation in tissue culture and directly in BAL cells. We found that PCR detected all cases positive by virus isolation (15/52 samples) and the result was obtained within 5 h. PCR detected more cases of CMV than did virus isolation (22/52 samples). Positive PCR and negative virus isolation were consistent with probable CMV infection in 3/7 patients when other clinical and laboratory parameters of CMV infection were considered. The negative predictive value of PCR was high; none of 30 patients negative by PCR developed CMV pneumonia within the subsequent 2 months. Detection of CMV antigen after short-term incubation was rapid enough to be used in clinical practice, specific (100%) and with a sensitivity of 60%. Demonstration of CMV antigen in alveolar cells was highly specific (100%) but had too low a sensitivity (26.7%) to be used as the only rapid method. Our conclusion is that a combination of PCR and detection of CMV antigen after short-term incubation and directly in alveolar cells is optimal for rapid identification of CMV. PMID- 8248741 TI - Chlamydia pneumoniae in Swedish patients. AB - Chlamydia pneumoniae infections have earlier been described as mycoplasma-like illnesses in young people, and also appear to be associated with community acquired pneumonia in adults. In this retrospective study, 12.2% (23/188) of patients with pneumonia who required hospitalization during the 3 years 1985-87 had serological evidence of recent C. pneumoniae infection. Many of these patients had symptoms similar to ornithosis. The most interesting finding was that half of the patients with a 4-fold IgG antibody titre rise to C. pneumoniae also had an increased alkaline phosphatase concentration. PMID- 8248742 TI - White blood cell and differential counts in acute respiratory viral and bacterial infections in children. AB - White blood cell (WBC) and differential counts were studied in 201 children hospitalized for acute viral or bacterial respiratory infection. The aetiology of infection was studied with a comprehensive set of serological tests. WBC and granulocyte counts were higher in patients with bacterial infection than in those with viral infection. Lymphocyte counts, by contrast, had no such aetiological association. The 95% confidence limits for WBCs and granulocytes distinguished bacterial and pneumococcal cases completely from viral cases with no bacterial involvement. The sensitivity of WBC counts, as well as granulocyte or lymphocytes counts, for distinguishing bacterial from viral cases was low at all cut-off levels. Specificity, in contrast, was 86% and 95% for WBCs at the cut-off levels 15.0 and 20.0 x 10(9)/l, and 84% and 97% for granulocytes at the cut-off levels 10.0 and 15.0 x 10(9)/l, respectively. It is concluded that high WBC and granulocyte counts are clear evidence of the bacterial aetiology of respiratory infection, but low or normal values do not rule it out. Lymphocyte counts are of no value for distinguishing between viral and bacterial infections. PMID- 8248743 TI - Monoclonal antibodies for identification of Borrelia afzelii sp. nov. associated with late cutaneous manifestations of Lyme borreliosis. AB - Borrelia isolates associated with Lyme borreliosis were previously divided into 3 genospecies, B. burgdorferi sensu stricto, B. garinii and group VS461, on the basis of DNA homology. B. burgdorferi sensu stricto and B. garinii were identified by monoclonal antibodies (MAbs), H3TS and D6 respectively, but no MAbs were available to identify group VS461. Two MAbs were produced, I 17.3 and J 8.3 which reacted with OspB and OspA proteins, respectively, of strains belonging to group VS461, which should be named B. afzelii sp. nov. 24 strains were assigned to B. afzelii sp. nov., 11 of them being isolated from skin lesions, 6 from acrodermatitis chronica atrophicans (ACA) and 5 from erythema chronicum migrans (ECM). Although quite unknown in the USA, ACA has frequently been reported in northern Europe where B. afzelii sp. nov. is commonly isolated. This study documents the involvement of B. afzelii sp. nov. as a specific aetiological agent of ACA. PMID- 8248744 TI - Seasonal prevalence of Borrelia burgdorferi in Ixodes ricinus in different vegetation types in Sweden. AB - The aim of this investigation was to estimate the seasonal risk of contracting human Lyme disease in different vegetation types in southern Sweden. Host-seeking Ixodes ricinus ticks (Acari: Ixodidae) were collected with standardized methods during May-September 1988 and March-October 1989 at 10 different sampling sites. Tick abundance was greatest during May-June and August-September. Prevalence of Borrelia burgdorferi infection of the ticks was assessed by phase-contrast microscopy complemented by immunology. Spirochetal infection of tick larvae was not detected. The prevalence of infection among nymphal ticks differed significantly between years and between sampling sites. Infection prevalence was greater in adult females than in nymphs, but was similar in female and male ticks. Among all vegetation types studied, the greatest Lyme disease risk was deemed to be from I. ricinus nymphs during May and September in mixed forest vegetation. PMID- 8248745 TI - Evaluation of DNA-DNA hybridization for the direct detection of enterotoxigenic Escherichia coli in stool blots. AB - The simplicity of enterotoxigenic Escherichia coli (ETEC) stool blot hybridization, where the total bacterial growth of a fecal inoculum is examined directly for the presence of enterotoxin genes, has been marred by reports of unsatisfactory sensitivity and/or specificity. To assess the accuracy of stool blot hybridization and to study the effect of varying proportions of ETEC among fecal E. coli (ETEC/E. coli) on test performance, a detailed 'blind' study of 166 stool specimens from children with diarrhea was performed. Oligonucleotide probes were found to be superior to polynucleotide probes, having a sensitivity of 80%, a specificity of 99%, a positive predictive value of 89% and a negative predictive value of 97%. The sensitivity was found to be 100% when ETEC/E. coli > 2/12, as compared with 20% when ETEC/E. coli < or = 2/12 (p = 0.001), showing that the proportion of ETEC among fecal E. coli is of paramount importance for test sensitivity. PMID- 8248746 TI - Epidemiological aspects of community-acquired and hospital-acquired bacteremia in northern Norway. AB - 389 patients with 401 bacteremic episodes, either community-acquired (CAB) or hospital-acquired (HAB), admitted to the University Hospital of Tromso (UHT), Norway, and 3 small local hospitals in the region (LHs), were reviewed on the basis of data collected in 1985 and 1989. As regards incidence, sex- and age distribution, distribution to either surgical, medical, or pediatric wards, compared with etiological agents and predisposing factors, the statistical significance has been evaluated. The proportion of patients < 50 years was greater at UHT than in the LHs. More patients with HAB associated with predisposing factors were hospitalized at UHT than at the LHs. Coagulase-negative staphylococci occurred less frequently in bacteremic patients at UHT than at the LHs. As opposed to reports from elsewhere, Tromso University Hospital seems to appear much more similar to the smaller local hospitals in the epidemiological aspects of bacteremia, inasmuch as both HAB and the CAB presented an even distribution pattern between the 2 hospital groups. PMID- 8248747 TI - Effects of imipenem, cefotaxime and cotrimoxazole on aerobic microbial colonization of the digestive tract. AB - Superinfections originating from a digestive tract colonized by abnormally high concentrations of aerobic microorganisms as a result of impaired resistance to colonization (CR) may complicate antibiotic therapy. In this study, patients with a moderate to severe systemic infection were randomized to receive either cefotaxime (CTX, n = 10) or cotrimoxazole (CTR, n = 10), 2 antibiotic regimens presumed to spare CR; or imipenem/cilastine (I/C, n = 19). The effect on CR was measured indirectly by comparing the aerobic faecal flora before antibiotic treatment with that on day 8 of treatment. An increase in aerobic faecal flora denotes a disturbed CR, whereas a decrease means that the organism is sensitive to the effective faecal concentration of the antibiotic. Imipenem/cilastine treated patients showed a significant increase in enterococci and Candida spp., while the number of aerobic Gram-negative rods remained constant. Cefotaxime treated patients had evidence of an increase in enterococci, but not of Candida spp., and Escherichia coli numbers decreased significantly. In these patients the concentration of other Gram-negative aerobic rods showed a slight increase in 6 patients with a resistant Pseudomonas strain. Cotrimoxazole-treated patients showed a significant decrease in aerobic Gram-negative rods, a significant increase in Candida spp. and no change in enterococci. It is concluded that all 3 antimicrobial agents impair colonization resistance. Whether or not this is followed by overgrowth with resistant micro-organisms depends on the active faecal concentration of the antimicrobial agent and the MIC of the aerobic micro organisms. The risk of overgrowth of the bowel with resistant Gram-negative bacilli appears to be smaller following cotrimoxazole than following cefotaxime or imipenem/cilastine. PMID- 8248748 TI - Early Candida isolations in febrile patients after abdominal surgery. AB - The significance of finding Candida sp. in febrile patients after abdominal surgery was studied in a prospective study on 107 patients. The patients were divided into 3 groups, based on the isolation and time of sampling of Candida sp. and on any other microbiological documentation of the infection. Eight patients were found Candida-positive either by blood cultures or by cultures of urine, abdominal pus, tracheal aspirate or wound exudate within the first postoperative week and had significantly greater mortality than patients with other infections or later Candida sp.-isolation (50% vs 10%, p < 0.05). Isolation of Candida was associated with prolonged antibiotic therapy, extended central vein catheterization, parenteral nutrition and operations on the small bowel, ascending colon or pancreas, but not with the underlying illness. The results support the view that isolation of Candida sp. within the first week after surgery does not represent harmless colonization but is rather associated with serious morbidity. PMID- 8248749 TI - Clinical experience with multilamellar liposomal amphotericin B in patients with proven and suspected fungal infections. AB - Over a 3-year period, an unsonicated multilamellar vesicle preparation containing a low ratio of amphotericin B (5 mole %) was used as a routine alternative to amphotericin B-deoxycholate in treating 17 patients with a variety of systemic fungal infections representative of those commonly encountered on a tertiary care centre infectious disease service. Patient acceptability and convenience of administration were noteworthy. In 6/7 patients who had been given the liposomal drug after experiencing severe side effects (primarily hypokalemia and marked elevation of serum creatinine) on the non-liposomal form, the problems that had led to institution of the liposomal drug were reversed during treatment. However, multilamellar liposomal amphotericin B at conventional dosage was not without detectable toxicity in this patient population. Three transplant patients receiving cyclosporin at the same time as liposomal amphotericin B experienced a rise in serum creatinine, and 4 patients became hypokalemic during treatment: none of these effects was severe or required discontinuation of therapy. One or more liver enzymes rose measurably in 7 patients during treatment with liposomal amphotericin B, but remained unchanged or actually decreased in the remaining patients. PMID- 8248750 TI - Kawasaki disease in Finland in 1982-1992. AB - Data on patients suffering from Kawasaki disease have been collected prospectively in Finland since 1981. To ascertain the incidence, treatment and outcome in these patients the data were analysed for the 10-year period April 1982 to March 1992. Kawasaki disease was confirmed in 229 patients. The annual attack rate varied from 3.1 to 7.2 per 100,000 children below the age of 5 years. Their mean age was 3.9 years (range: 7 weeks to 39 years). 40% were under 2 years and 71% under 5 years of age. There was 1 adult patient. The male/female ratio was 1.5:1. Coronary artery lesions were found in 28 patients, 2 of whom died of myocardial infarction. Since April 1985, 132 patients (73%) have been treated with intravenous immunoglobulin. When treatment was begun before day 7 of the disease, coronary artery lesions were found in 5.5%; when begun between days 7 and 10, in 12%, and when begun after day 10, in 28% of the patients. PMID- 8248751 TI - Involvement of dogs in plague epidemiology in Tanzania. Serological observations in domestic animals in Lushoto District. AB - Venous blood was collected aseptically from clinically healthy domestic dogs, goats, sheep, cats and fowl in various plague-infected villages of Lushoto District, Tanzania, at the time when the disease was actively prevalent in the area. Flea ectoparasites were collected from the animals, processed, identified and counted. Serum samples were tested for specific plague antibodies, using the passive haemagglutination technique and checked by passive haemagglutination inhibition tests. Altogether 389 animals, of which 201 were domestic dogs, were involved. 11 (5.5%) dogs had significantly elevated specific plague antibodies at titres ranging from 20 to 1280. All the dogs were also heavily infested with fleas at a mean index of 7.7 fleas per animal. Of 1,871 fleas collected from the dogs, 93.8% were Ctenocephalides felis and 6.2% were C. canis. All the other animals examined were negative for plague. It was concluded that domestic dogs could play an important role as plague carriers in the area and that the animals could serve as sentinel animals for the detection of plague in villages where human plague outbreaks have not previously occurred. PMID- 8248752 TI - A study of the epidemiology of an endemic strain of staphylococcus haemolyticus (TOR-35) in a neonatal intensive care unit. AB - Coagulase-negative staphylococci (CNS) are among the most prevalent microorganisms that colonize and cause sepsis in neonatal intensive care units (NICU). We had previously identified a strain of CNS, Staphylococcus haemolyticus (TOR-35), in the NICU at Mount Sinai Hospital, that had been repeatedly isolated from blood cultures from neonates. We therefore carried out a prospective study to determine the frequency and time of colonization and the frequency of bacteremia in neonates over a 3.5 month period. This was accomplished by obtaining surface swabs within 1 h of birth and on days 3, 5, and 7 and by characterizing all blood culture isolates of CNS. We also determined what percentage of neonatal CNS bacteremias were due to this strain, between January 1, 1987 and December 31, 1990, by retrieving and typing all stock cultures of CNS from that period. All isolates were typed by species identification and antimicrobial susceptibility profile code. There were 76 (38%) neonates that became colonized with the TOR-35 strain at some time during their NICU stay. Lower birth weight was associated with colonization (p < 0.001), as was lower gestational age (p < 0.001). Only 1 neonate had a positive blood culture isolate for the TOR-35 strain during the prospective study. Of the 4 years of neonatal bacteremias that were studied retrospectively, there were 252 episodes of CNS bacteremia, of which 27 (11%) were due to the TOR-35 strain. The TOR-35 strain has become endemic in our NICU and appears to selectively colonize premature, low birth weight newborn infants, but only infrequently causes bacteremia. PMID- 8248753 TI - IgM response to a human Pneumocystis carinii surface antigen in HIV-infected patients with pulmonary symptoms. AB - We have developed an ELISA to detect IgM antibodies to a major human Pneumocystis carinii surface antigen (gp95), and investigated the IgM response in 128 HIV infected patients who underwent bronchoscopy for evaluation of pulmonary symptoms. Only 5 (4%) patients had IgM antibodies to P. carinii gp95. Four of the 5 patients with IgM antibodies also had IgG antibodies to gp95 and microbiologically proven P. carinii pneumonia (PCP). In 76/128 patients for whom serial samples were available, changes in antibody response were determined. In 3 patients we demonstrated an increase in IgM antibody response to gp95. These patients also showed an increase in IgG antibodies to gp95 and had microbiologically proven PCP. Prior to the development of the IgM response, IgG antibodies to gp95 were detectable in all 3 patients. Thus, HIV-infected patients with PCP seldom produce IgM antibodies to the major human P. carinii surface antigen. The increase in IgM response during the course of PCP observed in 3 patients suggests either reinfection with a new strain, or antigenic drift of an already acquired strain of P. carinii. PMID- 8248754 TI - Natural killer cell activity in herpes zoster in children without underlying disease. AB - We determined natural killer cell (NKC) activity in 10 otherwise normal children with herpes zoster. NKC activity values in children with vs. without varicella zoster virus specific IgM antibodies and in controls were 28.3 +/- 8.6%, 11.9 +/- 3.6% and 20.2 +/- 3.8% (mean +/- SD), respectively. There were significant differences between the children with and without IgM antibodies during the acute phase (p < 0.005) and between children without IgM antibodies and controls (p = 0.005). NKC values in children with mild vs. moderate morbidity were 11.7 +/- 4.1% and 25.7 +/- 9.9%, respectively (p < 0.05). The morbidity was moderate in all children with IgM antibodies, but in only 2 of the 5 children without IgM antibodies. Children who contracted varicella when a few months old had the highest IgM antibody titers and the highest value of NKC activity. NKC activity was related both to the presence of IgM antibodies and to the morbidity of herpes zoster. PMID- 8248755 TI - Puumala virus infection without signs of renal involvement. AB - Five cases of Puumala virus infection without renal involvement are described. The main signs were fever, headache, chills and thirst. Pulmonary involvement was also common. Neither increased levels of serum creatinine nor abnormal urinalyses were detected. We suggest that Hantavirus infection is included in the differential diagnosis of any febrile condition when exposure to aerosols from infectious secretions or secretions from rodents is suspected. PMID- 8248756 TI - Ureaplasma urealyticum cultured from brain tissue of preterm twins who died of intraventricular hemorrhage. AB - We present a case of premature twins, born at 24 weeks of gestation. Both infants died of intraventricular hemorrhage, aged 1 and 3 days, respectively. Ureaplasma urealyticum was isolated from brain tissue obtained at the autopsy of both infants. Our observations lend additional evidence of the role of U. urealyticum as a central nervous system pathogen in premature infants. PMID- 8248757 TI - Imported relapsing fever in European tourists. AB - Two 'imported' cases of relapsing fever after a trip through Senegal are described. Two women developed a tick-borne relapsing fever after having slept outdoors on a terrace in Zinguichor, Senegal. The first patient was rapidly cured after a course of doxycycline. The second patient initially received erythromycin, but despite this treatment she developed neurological symptoms and Borrelia persisted in the thick-smear examination. After treatment with doxycycline she developed a Jarish-Herxheimer reaction. Treatment with doxycycline was continued and finally all symptoms disappeared within 36 hours after starting this treatment. A diagnosis of relapsing fever should be considered in all patients returning from the tropics with recurrent fever, especially if no malaria parasites are found. PMID- 8248758 TI - Low natural killer cell function in disseminated aspergillosis. AB - We investigated whether immune deficiency was associated with a fatal case of invasive cerebral aspergillosis due to Aspergillus fumigatus infection. The lymphocyte proliferative capacity to T- and B-cell mitogens was comparable to that of healthy controls. However, the natural killer (NK) cell activity of the patient was 2-6-fold lower than the activity expressed by a paired control and age-sex matched healthy controls (n = 20). No specific abnormalities were evident in the capacity to transduce either proliferative signals or cytokine (interleukin 2) secretory capacity of in vitro activated lymphocytes from the patient. The specific reduction in NK cell activity might either indicate that she was a low NK responder, or that the low NK cell activity was primarily or secondarily associated with the fungal infection caused by Aspergillus fumigatus in humans. PMID- 8248759 TI - Ketoconazole treatment of a family with zoonotic sporotrichosis. AB - All 3 members of a family developed lymphocutaneous and cutaneous sporotrichosis after exposure to an infected cat. The lesions resembled acute bacterial lymphadenitis and lymphangitis and responded within 2 weeks to ketoconazole treatment with complete healing and no recurrence over the next 6 months. Immunologically normal children and adults with lymphocutaneous or cutaneous sporotrichosis may respond well to ketoconazole therapy. PMID- 8248760 TI - Late onset polio sequelae in Denmark. Results of a nationwide survey of 3,607 polio survivors. PMID- 8248761 TI - Stroke rehabilitation: identification of target groups and planning data. AB - In order to ascertain the need for late stroke rehabilitation in South-West Stockholm (SWS) and explore the conditions for experimental stroke outcome research, we followed a population-based, systematic sample of incident stroke patients, hospitalized during the period 1987-1988, using computer files and a telephone interview. The annual incidence of first-ever or recurrent transitory ischaemic attack (TIA) or acute stroke during the period was 2.83 per 1000. In December 1989, approximately 3% of these incident patients declared subjective residual dysfunction, were living at home and were considered potential candidates for rehabilitation. The estimated prevalence of home-bound individuals with residual dysfunction due to stroke was 0.74 per 1000 (95% CI 0.51-1.04). The average duration of hospitalization for these patients was 95.2 days. Approximately 3/4 of them were interested in further conventional rehabilitation. The group not interested in this therapy were mainly females and were older. They scored lower in personal ADL and mental activity, but similarly in motor function. We concluded that the prevalence need for rehabilitation after stroke in the SWS community amounted to at least 1/1000. Early rehabilitation in the community, linked to post-acute stroke care with reduction of hospital stay would be functionally and socially advantageous and cost-effective for more than 25% of the incident stroke cases. Stroke rehabilitation and clinical follow-up are closely related processes that should be evaluated simultaneously. PMID- 8248762 TI - A comparison of pressure pain thresholds in different tissues and body regions. Long-term reliability of pressure algometry in healthy volunteers. AB - Pressure pain thresholds (PPTs) were measured in 12 healthy female volunteers with a hand-held electronic pressure algometer (Somedic). The PPTs over 30 points, mainly located on the trunk, were measured in a randomized order. The measurements were repeated after one week and again 10-13 weeks later. Three spots over nerve tissue had lower PPTs than nearby muscle. There were no consistent differences between muscle and periosteum within the same region. Overall there was a tendency for points in the nape region to have the lowest PPT, and those in the lumbosacral region to have the highest. The shoulder points had intermediate values. The interindividual differences were great. There was no difference between the mean PPTs from the first session and those from the second session. However, at the third session, 10 weeks later, the average PPT value was substantially higher than in the previous sessions. PMID- 8248763 TI - Pulmonary function and symptom-limited exercise stress testing in subjects with late sequelae of poliomyelitis. AB - Sixty-eight subjects, consecutively admitted to our rehabilitation hospital with a presumptive postpolio syndrome, were examined by pulmonary function and symptom limited exercise stress testing. The purpose of this investigation was to study how many of these subjects could be classified as suffering from cardiorespiratory deconditioning. The subjects had moderately reduced lung function of restrictive type, and none of the subjects had forced expiratory volume for one second (FEV1) below 30% of predicted value, indicating that hypoventilation would probably not occur. A pronounced reduction in maximal oxygen uptake (max VO2) was seen, especially in women. The maximal heart rate (max HR) values were above 70% of predicted values in all but one subject, indicating that the subjects might benefit from endurance training. Fifteen subjects had a suspected pulmonary limitation due to the exercise, with the ratio ventilation/maximal voluntary ventilation (V/MVV) above 70%. However, max HR in these subjects did not differ from that in the subjects with the ratio V/MVV below 70%. Thirteen other subjects had a ratio V/MVV < 70% but did not achieve respiratory quotient (R) > 1.0 and/or capillary lactate concentration > 4 mmol/l during exercise, indicating that muscular factors limited the exercise. These results indicate that cardiorespiratory deconditioning was considerable in most of our subjects with postpolio syndrome. PMID- 8248764 TI - Trapezius muscle pressure pain threshold and strain in the neck and shoulder regions during repetitive light work. AB - Perceived musculoskeletal discomfort caused by the repetition of a light work task was examined by rating the perceived musculoskeletal discomfort and measuring the muscle pressure pain threshold. Thirteen healthy female workers performed a repetitive light work task for 120 minutes. Based on the perceived musculoskeletal discomfort during the work task the subjects were assigned to one of two groups, i.e., those who reported most complaints and those who reported fewest complaints. The pressure pain thresholds of the trapezius muscles in the most-complaining group were significantly lower than those in the least complaining group. The pressure pain threshold was logarithmically correlated both with the perceived musculoskeletal discomfort and with the EMG signal amplitudes. PMID- 8248765 TI - Assessment of patients with chronic back pain using the "Functional Status Questionnaire". AB - Forty-eight patients with chronic back pain undergoing a rehabilitation program were assessed with the Functional Status Questionnaire (FSQ) during the first week of rehabilitation and after 6 weeks, 6 months and one year. In their first test, all the patients showed diminished functional capacity in at least one of five summary scales, indicating serious health problems and a need for rehabilitation. The test-retest correlation was k = 0.96, indicating high reliability of the questionnaire. After 6 weeks of rehabilitation the patients estimated significant improvements regarding "Satisfaction with health", "Mental health" and "Number of close friends". Some of these improvements were maintained after 6 months and one year. The "Basic ADL" and "Intermediate ADL" scales, however, showed no changes, indicating either low sensitivity or that no real changes had occurred during the rehabilitation period. The FSQ is recommended for use among patients with chronic back pain, but the tests need to be supplemented with other measurements concerning ADL capacity. PMID- 8248766 TI - Semi-quantitative SPECT scanning in acute ischaemic stroke. AB - We performed semi-quantitative SPECT scans using 99mTcm-HMPAO on 34 elderly subjects 10-15 days after ischaemic stroke. Each cerebral hemisphere was divided into five regions of interest. Asymmetry scores were determined for ten slices in each region using activity ratios to the other hemisphere and to the ipsilateral cerebellum. Outcome was assessed by maximal Barthel ADL score during the first two months after stroke. Asymmetry scores in the hemisphere involved by the stroke event were much higher than those in the opposite hemisphere. Subjects with poor outcome (maximal Barthel 0-12) had significantly higher asymmetry scores than those with good outcome (Barthel 13-20). The total asymmetry score (both hemispheres) and the score of the involved hemisphere predicted Barthel ADL score in a general linear model. Total asymmetry score remained a significant predictor of maximal Barthel score in multivariate models. Thus semi-quantitative SPECT scanning is of value in predicting functional recovery after stroke. PMID- 8248767 TI - Effectiveness of countermeasures applied in Belarus to produce milk and meat with acceptable levels of radiocaesium after the Chernobyl accident. AB - Countermeasures taken in Belarus to reduce transfer of radiocaesium from feeds to animals have combined live monitoring and monitoring of radiocaesium (137Cs and 134Cs) in bulk milk on dairy farms with the setting of upper limits for radionuclide contents for animal feeds. Only uncontaminated feeds have been used in the final fattening of animals before slaughter. Combined with widespread soil management programmes these measures have allowed the continued use of the majority of contaminated agricultural lands without substantial changes in farming practices. PMID- 8248768 TI - Chemical methods to reduce the radioactive contamination of animals and their products in agricultural ecosystems. AB - After major nuclear accidents the most effective action to reduce activity levels in animal products is to provide uncontaminated feed or feed diluted with uncontaminated material. However, uncontaminated feed may not always be available in such situations. Therefore other effective procedures are needed to reduce activity levels which must be easy to apply, low in cost and have no side effects on animals and their products. Chemical substances administered as feed additive to housed or grazing animals act by influencing gut absorption or increasing the rate of excretion from blood and organs. The main chemical methods used are: (1) dilution of radionuclides by stable isotopes or analogous inactive elements; and (2) natural or artificial binding agents. In this paper a summary and critical review of the different chemical methods described in the literature is given. PMID- 8248769 TI - Management methods of reducing radionuclide contamination of animal food products. AB - Examples are given of general management methods that will reduce the contamination of domestic food producing animals. Such methods will decrease the radiation exposure of the animal which could be of importance for its health and long-term production of food but more importantly in the short term they will decrease transfer of radioactive contamination to animal food products, i.e. milk, meat, offal and eggs. The most important radionuclides contaminating food products of animal origin, 90Sr, 131I and 137Cs, are specifically discussed in detail. By choosing proper general management methods it is possible to decrease the transfer of contaminating radionuclides to animal food products by an order of magnitude or more. PMID- 8248770 TI - Chemical methods for reduction of the transfer of radionuclides to farm animals in semi-natural environments. AB - The same chemicals can be used for reduction of radionuclide transfer to animals whether kept on farms or grazing in semi-natural and natural habitats. However, different techniques are required for administration of the active compounds. Dairy ruminants may be treated effectively by inclusion of chemicals in supplemental concentrates. Practical experience gained after the Chernobyl accident has shown that both clay minerals and hexacyanoferrates are effective in preventing high radiocaesium levels in animal products. Chemicals such as bentonite clays and CaCO3, used for reduction of 137Cs and 90Sr transfer respectively, must be fed in hectogram quantities and are only practical for dairy animals in semi-natural ecosystems. Salt licks and sustained release boli with hexacyanoferrates as caesium binders have been developed and used successfully after the Chernobyl accident for meat producing cattle, sheep and reindeer which graze freely for extended periods. Daily doses of 25-300 mg in sheep and 250-2000 mg in cows reduces 137Cs accumulation 2-10-fold. Binders for 90Sr have not been tested in grazing animals. Stable iodine could be provided in salt licks and indwelling rumen boli at rates required to block radioiodine uptake by the thyroid gland. Boli and salt licks are highly cost effective in reducing doses to man when compared to interdiction of food from farm animals. PMID- 8248771 TI - Management methods of reducing radionuclide contamination of animal food products in semi-natural ecosystems. AB - Countermeasures involving changes in management practices which are suitable for use in semi-natural ecosystems are described. Most of the relevant information relates to radio-caesium, but the applicability for some other radionuclides has also been evaluated. Improved live-monitoring techniques for radiocaesium, developed since the Chernobyl accident, allow the identification of animals whose contamination levels exceed the intervention limits, so that countermeasures can be effectively chosen and targeted. Generally the most effective approach for domesticated and semi-domesticated animals is either to remove contaminated animals from the affected area or to provide uncontaminated feed. Uncontaminated feed is given continuously for dairy animals or in the final fattening stages for meat-producing animals. The introduction of other effective changes in management practices, such as changing hunting seasons for game, or slaughtering at a time of year when the animals have been grazing on less contaminated herbage, depends on a good understanding of the behaviour of radionuclides in these ecosystems. In practice, the most effective countermeasures which can used to reduce radionuclide contamination of animals in semi-natural ecosystems will be obtained by a combination of both management changes and the use of chemical binders to prevent gut absorption. PMID- 8248772 TI - Countermeasures for reducing radioactive contamination of farm animals and farm animal products. AB - A summary is given of the range and effectiveness of different available countermeasures which can reduce the radiation dose arising from the radioactive contamination of food products from animals. The choice of which countermeasure is suitable for each particular situation will depend on many factors. Consideration should be given not only to the effectiveness of each countermeasure in terms of reduction of radionuclide content, but also to its practicality and cost, particularly where deposition levels are high or where high contamination levels in animal food products persist for a long period of time. Effective countermeasures which can reduce radiocaesium contamination of milk and meat from farmed animals by more than a factor of 10 have been developed, tested and used during the last 30 years. Less progress has been made in identifying effective binders for reducing levels of the other important contaminating radionuclides, such as I and Sr, in animal food products. PMID- 8248773 TI - Forests and forestry: radiation protection measures with special reference to the Chernobyl accident zone. AB - A large proportion of the area contaminated by the Chernobyl accident in the former USSR is forested and has presented unique problems when considering appropriate post-contamination management and clean-up techniques. These problems are related to the forest's role as both a source and sink for radioactive contamination. Although it has been suggested that resuspension from forested areas may provide a secondary source of contamination to adjacent land, data collected after the Kyshtym and Chernobyl accidents suggest that forest ecosystems may also be effective in limiting the further spread of contamination away from the point of initial deposition and that this effect will increase over time. Such evidence serves to highlight the importance of these ecosystems in influencing the behaviour of radionuclides immediately after their release to the environment. Management practices for forested areas adopted since 1986 are described and a critical appraisal is presented of engineering-based countermeasures implemented over the initial post-accident period. These were intended to remove large quantities of contaminated materials from the forest environment. However, it is suggested that the natural processes of self decontamination of trees and forest floor litter layers are sufficiently rapid and efficient to necessitate radical alterations to the technology based approaches adopted in the first 2 years after the Chernobyl accident. PMID- 8248774 TI - Basic research and the cost of health care. AB - Figure 4 (p. 997) of the article "The genesis and collapse of third millenium north Mesopotamian civilization" by H. Weiss et al. (20 Aug., p. 995) contained some errors. The correct figure appears below. [See Table in the PDF] PMID- 8248775 TI - Varmus: the view from Bethesda. Interview by Eliot Marshall. PMID- 8248776 TI - Dioxin tied to endometriosis. PMID- 8248777 TI - Proteins and organic solvents make an eye-opening mix. PMID- 8248778 TI - Claim of higher risk for women smokers attacked. PMID- 8248779 TI - A switch between two-, three-, and four-stranded coiled coils in GCN4 leucine zipper mutants. AB - Coiled-coil sequences in proteins consist of heptad repeats containing two characteristic hydrophobic positions. The role of these buried hydrophobic residues in determining the structures of coiled coils was investigated by studying mutants of the GCN4 leucine zipper. When sets of buried residues were altered, two-, three-, and four-helix structures were formed. The x-ray crystal structure of the tetramer revealed a parallel, four-stranded coiled coil. In the tetramer conformation, the local packing geometry of the two hydrophobic positions in the heptad repeat is reversed relative to that in the dimer. These studies demonstrate that conserved, buried residues in the GCN4 leucine zipper direct dimer formation. In contrast to proposals that the pattern of hydrophobic and polar amino acids in a protein sequence is sufficient to determine three dimensional structure, the shapes of buried side chains in coiled coils are essential determinants of the global fold. PMID- 8248780 TI - A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. AB - A DNA sequence rich in (A+T), located upstream of the -10, -35 region of the Escherichia coli ribosomal RNA promoter rrnB P1 and called the UP element, stimulates transcription by a factor of 30 in vivo, as well as in vitro in the absence of protein factors other than RNA polymerase (RNAP). When fused to other promoters, such as lacUV5, the UP element also stimulates transcription, indicating that it is a separate promoter module. Mutations in the carboxyl terminal region of the alpha subunit of RNAP prevent stimulation of these promoters by the UP element although the mutant enzymes are effective in transcribing the "core" promoters (those lacking the UP element). Protection of UP element DNA by the mutant RNAPs is severely reduced in footprinting experiments, suggesting that the selective decrease in transcription might result from defective interactions between alpha and the UP element. Purified alpha binds specifically to the UP element, confirming that alpha acts directly in promoter recognition. Transcription of three other promoters was also reduced by the COOH-terminal alpha mutations. These results suggest that UP elements comprise a third promoter recognition region (in addition to the -10, -35 recognition hexamers, which interact with the sigma subunit) and may account for the presence of (A+T)-rich DNA upstream of many prokaryotic promoters. Since the same alpha mutations also block activation by some transcription factors, mechanisms of promoter stimulation by upstream DNA elements and positive control by certain transcription factors may be related. PMID- 8248781 TI - Photoactivated conformational changes in rhodopsin: a time-resolved spin label study. AB - Rhodopsin has been selectively spin-labeled near the cytoplasmic termini of helices C and G. Photoactivation with a light flash induces an electron paramagnetic resonance spectral change in the millisecond time domain, coincident with the appearance of the active metarhodopsin II intermediate. The spectral change is consistent with a small movement near the cytoplasmic termination of the C helix and reverses upon formation of the MIII state. These results provide an important link between the optical changes associated with the retinal chromophore and protein conformational states. PMID- 8248782 TI - Rad: a member of the Ras family overexpressed in muscle of type II diabetic humans. AB - To identify the gene or genes associated with insulin resistance in Type II (non insulin-dependent) diabetes mellitus, subtraction libraries were prepared from skeletal muscle of normal and diabetic humans and screened with subtracted probes. Only one clone out of 4000 was selectively overexpressed in Type II diabetic muscle as compared to muscle of non-diabetic or Type I diabetic individuals. This clone encoded a new 29-kilodalton member of the Ras-guanosine triphosphatase superfamily and was termed Rad (Ras associated with diabetes). Messenger ribonucleic acid of Rad was expressed primarily in skeletal and cardiac muscle and was increased an average of 8.6-fold in the muscle of Type II diabetics as compared to normal individuals. PMID- 8248783 TI - Phosphatidylinositol 4-kinase: gene structure and requirement for yeast cell viability. AB - Phosphatidylinositol (PtdIns) 4-kinase catalyzes the first step in the biosynthesis of PtdIns-4,5-bisphosphate (PtdIns[4,5]P2). Hydrolysis of PtdIns[4,5]P2 in response to extracellular stimuli is thought to initiate intracellular signaling cascades that modulate cell proliferation and differentiation. The PIK1 gene encoding a PtdIns 4-kinase from the yeast Saccharomyces cerevisiae was isolated by polymerase chain reaction (PCR) with oligonucleotides based on the sequence of peptides derived from the purified enzyme. The sequence of the PIK1 gene product bears similarities to that of PtdIns 3-kinases from mammals (p110) and yeast (Vps34p). Expression of PIK1 from a multicopy plasmid elevated PtdIns 4-kinase activity and enhanced the response to mating pheromone. A pik1 null mutant was inviable, indicating that PtdIns4P and presumably PtdIns[4,5]P2 are indispensable phospholipids. PMID- 8248784 TI - The influence of antigen organization on B cell responsiveness. AB - The influence of antigen epitope density and order on B cell induction and antibody production was assessed with the glycoprotein of vesicular stomatitis virus serotype Indiana [VSV-G (IND)]. VSV-G (IND) can be found in a highly repetitive form the envelope of VSV-IND and in a poorly organized form on the surface of infected cells. In VSV-G (IND) transgenic mice, B cells were unresponsive to the poorly organized VSV-G (IND) present as self antigen but responded promptly to the same antigen presented in the highly organized form. Thus, antigen organization influences B cell tolerance. PMID- 8248785 TI - Mechanism-based inactivation of prostatic acid phosphatase. AB - Protein phosphatases play important roles in the regulation of cell growth and metabolism, yet little is known about their enzymatic mechanism. By extrapolation from data on inhibitors of other types of hydrolases, an inhibitor of prostatic acid phosphatase was designed that is likely to function as a mechanism-based phosphotyrosine phosphatase inactivator. This molecule, 4-(fluoromethyl)phenyl phosphate, represents a useful paradigm for the design of potent and specific phosphatase inhibitors. PMID- 8248786 TI - Actions of antiulcer drugs. PMID- 8248787 TI - Action of antiulcer drugs. PMID- 8248788 TI - Health care: more access and more cures. PMID- 8248789 TI - Medical research: alternative views. PMID- 8248790 TI - Dioxin exposure. PMID- 8248791 TI - Genetics and crime. PMID- 8248792 TI - A life-saving accelerator. PMID- 8248793 TI - Cyclic ADP-ribose and pancreatic beta cells. PMID- 8248794 TI - Skn-1a and Skn-1i: two functionally distinct Oct-2-related factors expressed in epidermis. PMID- 8248795 TI - Serious flaws in the horizontal approach to biotechnology risk. PMID- 8248796 TI - Biomedical funding. Prevention research: a new growth area for NIH? PMID- 8248797 TI - Race to grow hantavirus ends in tie. PMID- 8248798 TI - AIDS prevention trial seeks answers. PMID- 8248799 TI - Ribozymes: killing the messenger. PMID- 8248800 TI - New clues surface about the making of the mind. PMID- 8248801 TI - Enzymatic synthesis of a bacterial polyketide from acetyl and malonyl coenzyme A. AB - Microorganisms and plants manufacture a large collection of medically and commercially useful natural products called polyketides by a process that resembles fatty acid biosynthesis. Genetically engineered microorganisms with modified polyketide synthase (PKS) genes can produce new metabolites that may have new or improved pharmacological activity. A potentially general method to prepare cell-free systems for studying bacterial type II PKS enzymes has been developed that facilitates the purification and reconstitution of their constituent proteins. Selective expression of different combinations of the Streptomyces glaucescens tetracenomycin (Tcm) tcmJKLMN genes in a tcmGHIJKLMNO null background has been used to show that the Tcm PKS consists of at least the TcmKLMN proteins. Addition of the TcmJ protein to the latter four enzymes resulted in a greater than fourfold increase of overall activity and thus represents the optimal Tcm PKS. Polyclonal antibodies raised against each of the TcmKLMN proteins strongly inhibit the Tcm PKS, as do known inhibitors targeted to the active site Cys and Ser residues of a fatty acid synthase. This system exhibits a strict starter unit specificity because neither propionyl, butyryl, or isobutyryl coenzyme A substitute for acetyl coenzyme A in assembly of the Tcm decaketide. Because the Tcm PKS activity is significantly diminished by removal of the TcmM acyl carrier protein and can be restored by addition of separately purified TcmM to two different types of TcmM-deficient PKS, it should be possible to use such preparations to assay for each of the constituents of the Tcm PKS. PMID- 8248802 TI - Engineered biosynthesis of novel polyketides. AB - Polyketide synthases (PKSs) are multifunctional enzymes that catalyze the biosynthesis of a huge variety of carbon chains differing in their length and patterns of functionality and cyclization. Many polyketides are valuable therapeutic agents. A Streptomyces host-vector system has been developed for efficient construction and expression of recombinant PKSs. Using this expression system, several novel compounds have been synthesized in vivo in significant quantities. Characterization of these metabolites has provided new insights into key features of actinomycete aromatic PKS specificity. Thus, carbon chain length is dictated, at least in part, by a protein that appears to be distinctive to this family of PKSs, whereas the acyl carrier proteins of different PKSs can be interchanged without affecting product structure. A given ketoreductase can recognize and reduce polyketide chains of different length; this ketoreduction always occurs at the C-9 position. The regiospecificity of the first cyclization of the nascent polyketide chain is either determined by the ketoreductase, or the chain-extending enzymes themselves. However, the regiospecificity of the second cyclization is determined by a distinct cyclase, which can discriminate between substrates of different chain lengths. PMID- 8248803 TI - Binding to DNA and the retinoblastoma gene product promoted by complex formation of different E2F family members. AB - The E2F family of transcription factors functions in the control of the mammalian cell cycle. Here it is shown that two family members, E2F-1 and DP-1, form specific heterodimers in vivo, a process that enhances DNA binding, transactivation, and the binding of the retinoblastoma gene product. These results suggest that heterodimerization regulates E2F function and contributes to cell cycle control. PMID- 8248804 TI - Visualization of single molecules of RNA polymerase sliding along DNA. AB - Transcription requires that RNA polymerase binds to promoters buried in nonspecific sites on DNA. The search for promoters may be facilitated if the polymerase slides along the molecule of DNA. Single molecules of Escherichia coli RNA polymerase were visualized, and their movements on immobilized bacteriophage lambda and T7 DNAs were examined. Deviating from drifts by bulk flow, about 40 percent of the enzyme molecules moved along the extended DNA. The results provide direct evidence for sliding as a mechanism for relocation of the enzyme on DNA. PMID- 8248805 TI - Nucleosome disruption by transcription factor binding in yeast. AB - Studies in vivo and in vitro have shown that the packaging of DNA into chromatin can affect gene expression. Here, binding of the yeast transcriptional activator GAL4 to DNA in chromatin has been investigated in vivo with a yeast episome. A positioned nucleosome that is present in cells grown in glucose and contains a single GAL4 binding site is disrupted by GAL4 binding in galactose. GAL4 can also bind to DNA in chromatin when the carboxyl-terminal activation domain of GAL4 is either masked by GAL80 or is absent. These results show that a transcription factor can bind to its site in vivo in what would appear to be a repressive chromatin structure. PMID- 8248806 TI - Tethering ribozymes to a retroviral packaging signal for destruction of viral RNA. AB - Cellular compartmentalization of RNAs is thought to influence their susceptibility to ribozyme cleavage. As a test of this idea, two retroviral vectors--one encoding a hammer-head ribozyme designed to cleave lacZ transcripts and another encoding the lacZ messenger RNA--were coexpressed inside retroviral packaging cells. Because of the retroviral packaging signal, the ribozyme would be expected to colocalize with the lacZ-containing viral genomic RNA but not with the lacZ messenger RNA. The ribozyme was found to reduce the titer of infectious virus containing lacZ by 90 percent, but had no effect on translation of lacZ messenger RNA. These results indicate that sorting gene inhibitors to appropriate intracellular sites may increase their effectiveness. PMID- 8248807 TI - A link between cyclin A expression and adhesion-dependent cell cycle progression. AB - Cell adhesion has an essential role in regulating proliferation during the G1 phase of the cell cycle, and loss of this adhesion requirement is a classic feature of oncogenic transformation. The appearance of cyclin A messenger RNA and protein in late G1 was dependent on cell adhesion in both NRK and NIH 3T3 fibroblasts. In contrast, the expression of Cdc2, Cdk2, cyclin D1, and cyclin E was independent of adhesion in both cell lines. Transfection of NRK cells with a cyclin A complementary DNA resulted in adhesion-independent accumulation of cyclin A protein and cyclin A-associated kinase activity. These transfected cells also entered S phase and complete multiple rounds of cell division in the absence of cell adhesion. Thus, cyclin A is a target of the adhesion-dependent signals that control cell proliferation. PMID- 8248808 TI - Involvement of platelet-endothelial cell adhesion molecule-1 in neutrophil recruitment in vivo. AB - During inflammation, neutrophils migrate from the vascular lumen into extravascular sites. In vitro assays have suggested that platelet-endothelial cell adhesion molecule-1 [PECAM-1 (CD31)], a member of the immunoglobulin superfamily, is required for the transmigration of neutrophils across endothelial monolayers. Antibody to human PECAM-1, which cross-reacts with rat PECAM-1, was found to block not only in vivo accumulation of rat neutrophils into the peritoneal cavity and the alveolar compartment of the lung but also neutrophil accumulation in human skin grafts transplanted onto immunodeficient mice. On the basis of these findings in three different models of inflammation, it appears that PECAM-1 is required for neutrophil transmigration in vivo and may thus be a potential therapeutic target. PMID- 8248809 TI - T cell receptor specificity and diabetes in nonobese diabetic mice. PMID- 8248810 TI - T cell receptor specificity and diabetes in nonobese diabetic mice. PMID- 8248811 TI - Effect of platelet-associated virus on assays of HIV-1 in plasma. PMID- 8248812 TI - Scintigraphy of spinal disorders in adolescents. AB - Bone scintigraphy in adolescents is useful in helping to differentiate between developmental (atypical lumbar Scheuermann disease), infectious (discitis, osteomyelitis), neoplastic (osteoid osteoma, osteoblastoma), and traumatic (occult fractures, spondylolysis, pseudoarthrosis) disease of the spine. Double phase (blood pool, delayed images) scintigraphy can characterize the pattern (i.e., linear in fracture, ovoid in nidus of osteoid osteoma). Single-photon emission computed tomography (SPECT) can be helpful in detecting the subtle presence of stress reaction (spondylolyses) not noted on routine planar scintigraphy and radiography. Bone scintigraphy is most beneficial when correlated with other imaging modalities in refining the diagnosis of spinal diseases. PMID- 8248813 TI - Magnetic resonance imaging of patellofemoral relationships. AB - Patellofemoral relationships were analyzed in 11 patients (13 knees) with patellar dislocation and 15 asymptomatic subjects (15 knees) at 0 degree and 20 degrees of flexion. The measurements were made from five consecutive axial images through the patellofemoral joint. The six indices measured were lateral patellar tilt (LPT), lateral patellofemoral angle (LPA), lateral patellar displacement (LPD), patella-lateral condyle index (L/PW), congruence angle (CA), and sulcus angle (SA). The reproducibility of the method was evaluated. The difference between the two study groups was more evident at 0 degree than at 20 degrees of knee flexion. Significant differences were noted between measurements made at different levels of the joint, particularly in the controls. Isometric contraction of the quadriceps muscle lateralized and tilted the patella slightly in both groups. L/PW with and without quadriceps muscle contraction, and LPA with reference to the anterior condyles differentiated between the two study groups most clearly. LPT and LPA with reference to the anterior condyles differentiated the study groups better than LPT and LPA with reference to the posterior condyles. The reproducibility was good except for inter-observer comparison of CA and SA. The use of an imaging plane selected at the midpoint of the patellar articular cartilage increases the sensitivity of the measurements, since it takes into account both the height of the patella and the tendency towards lateralization. These results indicate that patellar tilt is best measured with the LPA index and patellar lateralization with the L/PW index at 0 degree knee flexion. This study should always include isometric contraction of the quadriceps muscle. PMID- 8248814 TI - The patellar tendon: thickening, internal signal buckling, and other MR variants. AB - We studied the range of appearance of asymptomatic patellar tendons and evaluated the effect of age, weight, joint effusions, and anterior cruciate ligament (ACL) tears on this tendon. One hundred and seventy-three patellar tendons in asymptomatic patients were studied at 1.5 tesla. Sagittal short and long TE images were evaluated in regard to tendon thickness, ratio of thickness of patellar to quadriceps tendons, frequency, location, and severity of intratendon signal, and frequency and severity of tendon buckling. Results were correlated with patient age, sex, weight, the presence of ACL tears, and relative volumes of joint fluid. The mean thickness of the patellar tendon was 0.52 cm. The patellar to quadriceps tendon ratio was 0.72. The patellar tendon frequently (74%) had focal areas of signal apparently within it. This signal was usually subtle, V shaped (95%), and seen posteriorly in the proximal end of the tendon (82%). Intratendon signal was also seen commonly in the inferior aspect of the tendon (32%). This signal intensity did not increase with greater T2-weighting (99%). Buckling of the patellar tendon was a frequent asymptomatic variant (71%) but was also associated with joint effusions (p < 0.01) and ACL tears (p = 0.01). Buckling, intratendon signal, and tendon thickness increased with weight and age. Variation of the magnetic resonance appearance of the patellar tendon is frequent. Many of these changes appear to represent subclinical degeneration. Buckling of this tendon also may occur secondary to joint effusions or ACL tears. PMID- 8248815 TI - Paranasal sinus osteomas: a review of 46 cases. AB - In a survey of 1500 coronal sinonasal CT scans, 46 patients were found to have paranasal sinus osteomas. The overall incidence was 3%, with a predilection for decades 5 and 6 and a male-to-female ratio of 1.3:1. The frontal sinus was most commonly involved, and of these lesions 37% were in the immediate vicinity of the nasofrontal duct and 21% above and lateral to the ostium. Tumour size varied from a mean diameter of 1.5 mm to 30 mm; however, 17 lesions were 5 mm in size or less. Five matrix patterns were observed. Only two patients were symptomatic and only three osteomas excised. Isotope bone scanning may be a useful screening test with specific follow-up utilising computed tomography. PMID- 8248816 TI - An absence of structural changes in the proximal femur with osteoporosis. AB - The hypothesis that osteoporosis occurs not as a preferential loss of the tensile trabeculae but as a general loss of bone was tested by using bone mineral densitometry and an indentation test on dissected proximal femora. As osteoporosis advanced a significantly correlated decrease was found in both bone mineral density and mechanical properties between the principal compressive and tensile trabeculae. The decrease correlated with a decrease in the Singh index. These findings led to the conclusion that a sequential bone loss from the tensile trabeculae to the compressive ones did not occur as Singh reported, but instead a generalized loss of bone mineral in both the tensile and compressive trabeculae supervened. The structural changes, on which the grading system by Singh was based, were not observed in the proximal femur affected by osteoporosis. PMID- 8248817 TI - Bone metastases as the presenting manifestation of rhabdomyosarcoma in childhood. AB - Rarely, rhabdomyosarcoma can present with bone pain and bone lesions on radiographs without evidence of a primary tumor. Of 428 children with biopsy proven rhabdomyosarcoma, four presented with radiographic evidence of bone metastases, but no primary tumor was found on subsequent evaluation. On radiographs, these metastases, located most commonly in the metaphyses of the extremities and in the spine, displayed a destructive or diffusely permeative pattern without sclerotic margins and mimicked the more common neuroblastoma. One patient also had diaphyseal cortical lytic metastases of the tibia. Radiographs defined metastases of the extremities better than the correlative bone scans. In the spine, on T2-weighted magnetic resonance (MR) images, metastases displayed high signal intensity which contrasted with the low-signal-intensity marrow in these pediatric patients. On histopathologic examination, metastatic rhabdomyosarcoma was composed of small cells of variable size, shape, and growth pattern similar to other round cell tumors. A positive desmin immunohistochemical test helped to establish the diagnosis. The radiologist, pathologist, and clinician should be aware of this unusual presentation of rhabdomyosarcoma so that suitable immunohistochemical tests are performed and appropriate chemotherapy given. PMID- 8248818 TI - CT evaluation of primary epiphyseal bone abscesses. PMID- 8248819 TI - Case report 784: Fibrous dysplasia of the second pedal digit. AB - In summary, we have reported on an unusual form of fibrous dysplasia, presumptively monostotic, localized to the middle phalanx of the second pedal digit, and presenting as a mass. Monostotic fibrous dysplasia of phalanges of the foot is rare, but should be considered in the differential diagnosis of a mass in the foot. PMID- 8248820 TI - Case report 798: Recurrent chondromyxoid fibroma in connective tissue of leg. AB - In summary, this is a case of recurrent CMF in soft tissue, 17 years after the diagnosis of the initial lesion. The literature concerning this entity is reviewed and the histological and radiological patterns are considered. The causative factors are evaluated and the prognosis is discussed. PMID- 8248821 TI - Case report 799: Extraskeletal chondrosarcoma (Fig. 3). AB - In summary, extraskeletal chondrosarcoma is a rare soft tissue tumor. There may or may not be calcification, and in uncalcified tumors the diagnosis may not even be considered by the radiologist. The body of information as to clinical outcome is limited. Both of the patients whose tumors have been illustrated here are alive 2 years following surgery. PMID- 8248822 TI - Case report 800: Skeletal fibrous dysplasia associated with intramuscular myxoma (Mazabraud's syndrome). AB - In summary, intramuscular myxoma associated with fibrous dysplasia of bone represents a benign disorder of uncertain etiology. Magnetic resonance imaging has proved to be a useful diagnostic tool in the evaluation of this benign disorder. In addition, MR is a valuable aid in the preoperative planning process. PMID- 8248823 TI - Case report 801: Osteoid osteoma of the coccyx. AB - In summary, a typical osteoid osteoma in the coccyx in a 13-year-old boy has been presented-a unique location. The clinical, radiological, and pathological aspects of osteoma are summarized. The relationship of osteoid osteoma to osteoblastoma is stressed; it is also stressed that the occurrence of osteoid osteoma in the coccyx is most unusual. PMID- 8248824 TI - Case report 802: Ewing's tumor of the proximal phalanx of the third finger with radiographic progression documented over a 6-year-period. AB - In summary, a case of Ewing's tumor of the proximal phalanx of the long finger of the right hand was presented. This case is unusual because of its location as well as the unusually long clinical course before the correct diagnosis was made. The major point to be stressed is that Ewing's tumor may initially present with a "benign" appearance mimicking a bone cyst [17] or angioma [12]. Timely follow-up of so-called benign lesions, particularly when recurrent swelling occurs, would help in avoiding prolonged delays in diagnosis. PMID- 8248825 TI - Case report 803: Gorham's disease (Gorham-Stout syndrome) of scapula. AB - In summary, a 36-year-old man presented with pain and limited motion in the shoulder. Clinical examination revealed obliteration of normal scapular landmarks in the conspicuous absence of any palpable soft tissue mass. Roentgenograms showed progressive osteolysis of the scapula. Biopsy confirmed the diagnosis of Gorham's disease. MRI played a key role in defining the extent of disease involvement and in displaying the distinct soft tissue anatomy. These MRI features are to our knowledge previously undescribed. PMID- 8248826 TI - Case report 804: Chondroblastic osteosarcoma grade 3 of the left clavicle. AB - The summary, the case of a 15-year-old boy with osteosarcoma of the clavicle is presented, prompted by the rarity of primary tumors at that site and by this particular tumor's unusual roentgenographic appearance, which suggested a benign process. A review of the literature on clavicular tumors and malignancies is provided. Though extremely rare, primary tumors of the clavicle are more likely to be malignant than benign. This suggests tht serious consideration should be given to malignancy in the face of an apparently aggressive lesion of the clavicle, and perhaps even a benign-appearing tumor as well. PMID- 8248827 TI - Case report 805: Synovial sarcoma, epithelial hyperplasia in the humeral air sacs, and formation of medullary bone. AB - In summary, we have reported a most interesting case of synovial sarcoma, epithelial hyperplasia in the air sacs, and formation of medullary bone in the left humerus of a 5-year-old female white pigeon. The tumor showed aggressive features radiologically and caused dysfunction of the left wing. The histologic findings and immunohistochemical reactions in this report were identical in many respects to the manifestation of synovial sarcoma described in human beings and dogs. Formation of medullary bone in female birds during the reproductive phase and hyperplasia of epithelia in the air sacs were also discussed. PMID- 8248828 TI - Origin and control of gastrointestinal motility. AB - As a result of improved understanding of the origin and control of motility at both the whole organ and the cellular level, a scientific approach to the diagnosis and treatment (both medical and surgical) of motility disorders has evolved. Examples are present for all levels of the gastrointestinal tract. Manometric, myoelectric, and pharmacologic studies have elucidated the role of the lower esophageal sphincter and stomach in the pathogenesis of gastroesophageal reflux and determined the mechanism of successful medical and surgical treatment. Better evaluation of colorectal motility using colonic transit studies, pelvic floor radiography, and rectoanal manometrics has led to a better identification of both the etiology of severe constipation and patients who will have a successful surgical outcome. Studies of normal and abnormal gallbladder motility and responsiveness to hormonal stimulation have shed light on the cellular abnormalities in gallbladder myocytes that predispose to gallstone formation. Finally, since we have learned that certain surgical procedures affect motility in an adverse manner, a better basic understanding of gastrointestinal physiology has led to a better clinical understanding of the mechanism by which the changes occur and to the development of more directed physiologic operations. The classic example is seen in ulcer surgery, where the introduction of highly selective vagotomy instead of truncal vagotomy preserved antral innervation and decreased the incidence of postvagotomy complications. All these concepts and more are addressed in more detail in subsequent articles in this issue. PMID- 8248829 TI - Esophageal physiology and pathophysiology. AB - This article presents the normal physiology of esophageal peristalsis. It discusses current approaches to the diagnosis and treatment of primary disorders of the esophagus, including achalasia, nutcracker esophagus, diffuse esophageal spasm, as well as the secondary disorder, scleroderma. PMID- 8248830 TI - Gastroesophageal reflux. AB - Gastroesophageal reflux is a common disease that accounts for approximately 75% of esophageal pathology. Motility abnormalities of the esophagus and stomach, including an incompetent lower esophageal sphincter, are responsible for pathologic reflux in the majority of patients. Surgical treatment offers the only chance for long-term cure. Obtaining optimal results following surgery of the esophagus is one of the most challenging aspects of modern surgical therapy. Given a precise diagnosis, careful patient selection, critical attention to detail in the perioperative period, and the meticulous performance of the appropriate anti-reflux procedure, long-term success can be assured in more than 90% of patients. PMID- 8248831 TI - Gastric motor physiology and pathophysiology. AB - The stomach has two distinct physiologic motor areas: the proximal stomach and the distal stomach. The proximal stomach, with its slow, sustained contractions, has a key role in regulating intragastric pressure and gastric emptying of liquids, while the distal stomach, with its peristaltic contractions, has a major role in mixing, trituration, and emptying of solids. Diseases and operations that disturb the motility of these two areas can result in unique adverse motor sequelae. For example, operations that impair proximal gastric motility, such as proximal gastric resection, may cause rapid gastric emptying of liquids and subsequent dumping and diarrhea. In contrast, operations that impair distal gastric contractions, such as truncal vagotomy, may cause slow gastric emptying of solids and chronic gastric atony. Knowledge of the physiology of the stomach in health and of the pathophysiology with disease and after operation provides a basis for the successful treatment and prevention of these disorders. PMID- 8248832 TI - Gastrointestinal pacing. AB - Gastrointestinal pacing has been used to alter motor function and is effective in animal models in modulating gastric emptying, intestinal transit, and absorption. Application of gastrointestinal pacing to disorders of the human stomach and small bowel is an attractive treatment option that may some day become a clinical reality. Several technical problems must be overcome before further clinical testing can proceed. PMID- 8248833 TI - Normal physiology of small intestinal motility. AB - The small intestine produces a number of different contractions in various spatial and temporal patterns that promote efficient digestion, absorption, and propulsion of ingested material. The small bowel also serves a protective role through the use of special situation contractions that rapidly propel enteric contents into the stomach or colon, from which they may be expelled. Contractile activity of the small intestine is coordinated by an interplay of myogenic, neural, and chemical controls. PMID- 8248834 TI - Pathophysiology of small intestinal motility. AB - Only in the last few years have we begun to understand the effects of surgical procedures on contractile activity of the small intestine. Although most patients readily adapt to the procedures that are performed, operations such as small intestinal transplantation make it imperative that we pursue a more thorough understanding of the effects of surgery on the motility of the small intestine. PMID- 8248835 TI - Colonic motor activity. AB - The colon exhibits three types of contractions: individual phasic (short and long duration), organized groups (MMCs and nonmigrating motor complexes), and ultrapropulsive (giant migrating contractions). The individual phasic contractions and the MMCs and nonmigrating motor complexes produce extensive mixing and kneading of fecal material and slow net distal propulsion. The GMCs produce mass movements and expel feces during defecation. All contractions are controlled by myogenic, neural, and chemical mechanisms. The myogenic mechanisms determine the timing and frequency of contractions and the duration and distance of propagation of contractions. The neurochemical mechanisms determine whether the contractions will occur at a given site. PMID- 8248836 TI - Pathophysiology of colonic motility disorders. AB - When no identifiable organic cause for colonic symptoms can be found, it is easy for the busy clinician to label the patient neurotic. It is evident that many of these "functional" disorders do reflect an underlying motility disorder, although our understanding is far from clear. However, currently, patients with severe constipation are evaluated in a much more rational manner and, as a consequence, are offered a reasonable therapeutic approach that can be predicted to have a good chance for success. We can hope that as our understanding of irritable bowel syndrome is strengthened, treatment will become more efficacious than the unproved and costly medications that are in use currently. Until dietary modification becomes commonplace, it is unlikely that the incidence of diverticular disease or its complications will change. Already, our understanding of ileus has allowed us to realize the benefits of laparoscopic surgery, and as our knowledge of the various gut hormones and the inhibitory role that some play in intestinal motility grows, ileus, and its resulting prolongation of hospital stay, may become less problematic. PMID- 8248837 TI - Anorectal motor physiology and pathophysiology. AB - Disorders of anorectal function are complex. A clear definition of the underlying pathophysiologic process is possible only with the use of the anal physiology laboratory. A reasonable treatment plan is then possible. PMID- 8248838 TI - Physiology and pathophysiology of gallbladder motility. AB - Most significant gallbladder disease is associated with gallbladder stasis. Gallbladder motility is controlled by a complex interplay of hormonal and neural factors. Experimental and clinical studies have demonstrated impaired motility in gallstone disease, and experimental evidence indicates that motility disturbances precede gallstone formation. The ability to measure gallbladder motility clinically has also resulted in better diagnosis and treatment for patients with chronic acalculous cholecystitis. PMID- 8248839 TI - Physiology of motor function of the sphincter of Oddi. AB - Human and experimental studies of the sphincter of Oddi have established that the sphincter is not a simple and passive smooth muscle portion of the biliary system; rather, it plays an active role in modulating bile flow into the duodenum in both the fasted and the postprandial states. The sphincter of Oddi in the opossum, and likely in man, demonstrates spontaneous phasic and perhaps peristaltic activity that affects bile flow into the duodenum. The sphincter appears to be under the control of a smooth muscle pacemaking-like region in the proximal sphincter that controls the frequency and direction of propagation of the phasic contractions. Immunohistochemical studies have documented the presence of dense concentrations of neuropeptide-containing myenteric nerves in the sphincter of Oddi. Physiologic studies have confirmed that these peptides, in combination with the classic gastrointestinal hormones, exert significant effects on biliary motility. Experimental studies of the motor physiology of the sphincter of Oddi have advanced our understanding of human biliary motility and dysmotility. A subset of patients experiences biliary-like pain after cholecystectomy. This pain may be attributable to either mechanical stenosis of the papilla or dysfunction of the sphincter of Oddi. No definitive test currently exists to establish the diagnosis with adequate sensitivity or specificity. It is hoped that further development of ultrasonographic, scintigraphic, manometric, or electromyographic techniques will allow a clearer definition of patients who truly have sphincter dysfunction and who might benefit from medical, endoscopic, or surgical therapy. PMID- 8248840 TI - Pathophysiology of the sphincter of Oddi. AB - Pathophysiology of the sphincter of Oddi--or sphincter of Oddi dysfunction- manifests as either a biliary-type pain syndrome or recurrent pancreatitis. Imaging studies are unreliable, and direct endoscopic manometry is used to diagnose this entity. Milwaukee biliary classification, in addition to manometry, helps guide therapy. Endoscopic sphincterotomy in selected patients achieves permanent relief of symptoms. Endoscopic therapy for recurrent pancreatitis is still experimental. PMID- 8248841 TI - Radiation therapy for early glottic carcinoma: indication for the wedge filter. AB - From 1977 to the end of 1987, 244 patients with early glottic carcinoma (T1N0M0) were treated with radiation therapy. For 95 patients (group A), a styrofoam head holder and band were used to ensure immobilization during treatment without a wedge filter and for 149 patients (group B) a shell fixing devise and wedge filter were used. Total radiation dose administered was 50 to 70 Gy over a period of five to seven weeks with fields of 5 x 5 cm (group A) and 5 x 5 cm (group B1: n = 74) or 6 x 6 cm (group B2: n = 75). The five-year relapse-free survival (RFS) rates for group A and B were 85% and 90%, respectively, and RFS was essentially the same (p = 0.241). In group B, RFS was also basically the same for groups B1 and B2 (p = 0.78). According to tumor size, however, in patients with large T1a lesions (total length of one vocal cord) five-year RFS rates for groups A and B were 62% and 88%, respectively, and RFS was statistically significantly different (p = 0.003). Up to five years, seven patients (9%) of group A and 23 patients (17%) of group B showed minor chronic complication. As a minor chronic complication, arytenoid edema was found more frequently in group B (n = 15) than group A (n = 0). Eleven of these 15 cases were treated with large field (6 x 6 cm). For large T1a lesions of glottic carcinoma, a wedge filter was useful. A small field (5 x 5 cm) with an appropriate angle of wedge filter is recommended to avoid late arytenoid edema and achieve a more homogeneous dose distribution. PMID- 8248842 TI - [Dosage specification at the ICRU reference point: the consequences for clinical practice. International Commission on Radiation Units and Measurements]. AB - In the Department of Radiotherapy, University of Tubingen, dose specification is performed according to the new ICRU recommendations since June, 1991. The purpose was to prescribe, record and report photon beam therapy in a consistent way applicable to most clinical situations and to provide uniform criteria for a comparison of treatment results from different radiotherapy centers. For "level 2" plans (including CT planning), the concept of dose specification has been changed to the "ICRU reference point" instead of to form the isodose encompassing the target volume as we did previously. A total of 24 representative treatment plans have been analyzed to assess the consequences of this change on dose prescription, treatment technique, and the size of the irradiated volume. Our results revealed that the prescription of single and total doses have to be individually adjusted according to the specific treatment plans. ICRU recommendations on dose homogeneity may significantly affect treatment technique and field sizes. Small deviations in dose prescription may be of particular influence on late responding normal tissues ("double trouble" due to simultaneous adjustment of single and total doses). Detailed comparisons of individual treatment plans with respect to physical, biological, and clinical data are obviously necessary in order to make sure that the outcome of a radiation treatment is not deviating too much from the clinical experience based on the former dose specification concept. PMID- 8248843 TI - [A mathematical description of the relative dosage distributions of stereotactic collimator tubes in 6-MV photon radiation]. AB - The relative dose distributions of small circular fields for 6 MV photons may be calculated using a simple mathematical model. This model has been developed for stereotactic collimators with cylindrical cross section and diameters between 5 mm and 30 mm. The model consists of the description of depth dose curves and off axis dose distributions. The function for off-axis dose distributions is calculated by convolution of a simple profile function with a constant function. The width of the constant function is defined by beam geometry. In the result of integration only the width of the profile function is unknown, but it can be iteratively calculated. The agreement between measured and calculated dose distributions was tested at 5 cm and 20 cm depth in water and 100 cm source-to surface distance using thermoluminescent dosimetry (TLD) and films, giving a resolution of 1 mm. The depth dose curve is described using the inverse square law and two exponential functions. The first of these functions contains the effective attenuation coefficient in the argument, the second describes the build up. The increase of irradiated volume with increase of field diameter is accounted for using scatter-air ratios. The calculations were compared with TLD and ion-chamber measurements. PMID- 8248844 TI - Second primary cancer in irradiated stage I testicular seminoma. AB - The incidence of second primary cancer (SPC) was determined in 64 irradiated stage I testicular seminoma patients, treated and followed at the Northern Israel Oncology Center from 1968 to 1988. Seven (11%) patients developed a total of eight second cancers. The cumulative risk for developing second primary cancer at ten, 15 and 20 years following the diagnosis of seminoma was 2.5%, 5.1% and 8.9%, respectively. Two patients developed SPC within the radiation field (urinary bladder, sigmoid colon) and four patients outside the radiation field (bronchogenic cancer, thymoma, malignant melanoma, thyroid cancer). In two patients, who developed lung cancer and testicular seminoma, respectively, scatter dose from the main radiation field could not be excluded as one of the factors contributing to the SPC. Three patients died as a direct result of their SPC. It is concluded that there is a low, but significant risk of solid SPC among patients apparently cured of early stage seminoma. It will be important in future surveillance studies of stage I seminoma to assess the risk of SPC in the non irradiated patients. PMID- 8248845 TI - Different immobilization procedures during irradiation influence the estimation of alpha/beta ratios in mouse lip mucosa. AB - In previous experiments an anaesthetic gas mixture of enflurane (Ethrane, Abbott, Belgium) and oxygen has been used in our department to position mice for local irradiation. The development of a new immobilization method allowed the investigation of possible dose modifying effects of enflurane with different gases as carrier on the radiation response of mouse lip mucosa. The effect of different immobilization procedures on single dose and fractionated irradiations was examined. The experiments were compared on the basis of the dose associated with 50% incidence of lower lip desquamation (ED50 +/- 95% CI). When mice were breathing air the ED50 value following single dose irradiation was 20.0 (19.6 to 20.5) Gy. For mice receiving an anaesthetic gas mixture of enflurane and oxygen the ED50 was decreased significantly to 17.5 (17.2 to 17.8) Gy, indicating a sensitizing effect of the anaesthetic mixture by a factor of 1.14. The impact of different experimental set-ups on the fractionation sensitivity has been assessed. In order to estimate alpha/beta ratios in different treatment conditions two and ten fraction experiments were performed, additionally to the single dose irradiations. Dose modification factors were 1.09 for two fractions and 0.97 for the ten fraction experiment. This varying dose modification leads to a pronounced shift in alpha/beta ratios (estimated by "direct analysis" and graphically illustrated with a Fe-plot). For air breathing animals an alpha/beta ratio of 16.4 (14.5 to 18.9) Gy was estimated by direct analysis, whereas for the enflurane/oxygen anaesthesia 8.7 (7.3 to 10.2) Gy were calculated. Both these data sets were adequately described with the linear-quadratic formalism, as was shown with Tucker's analysis. The sensitizing effect of oxygen alone in single dose irradiations (DMF: 1.07) suggests moderate physiological hypoxia in mouse lip mucosa. The present data emphasize the effect of different immobilization procedures on the estimation of alpha/beta ratios, especially when large fraction sizes are involved. Data from experiments involving large fraction sizes should be avoided in the estimation of repair parameters. Further experimental work has to be carried out to elucidate the possible dose modifying effects of anaesthetic agents in clinical practice, e.g. for infants and for patients undergoing intraoperative radiotherapy. PMID- 8248846 TI - [Mediastinal irradiation in small-cell bronchial carcinoma]. PMID- 8248847 TI - [Should radiotherapy be used in the intraocular metastases of solid tumors?]. PMID- 8248848 TI - [Radiochemotherapy in anal carcinoma: radiation plus 5-FU with and without long term cisplatin infusion]. PMID- 8248849 TI - Low density of dopamine D4 receptors in Parkinson's, schizophrenia, and control brain striata. AB - The purpose of this study was to determine whether dopamine D4 receptors could be detected in the human brain striatum by means of an indirect ligand-binding method, because no dopamine D4 receptor-selective ligand presently exists. The antipsychotic clozapine is more selective for the dopamine D4 receptor than for other dopamine receptors. Although most antipsychotic drugs act in the striatum to elicit Parkinson-like side-effects, clozapine is atypical in that it does not produce Parkinsonism. To understand this atypical action of clozapine, it would be helpful to know whether the presumed target for clozapine, the dopamine D4 receptor, is or is not present in the human striatum. We measured dopamine D4 receptors indirectly, using [3H]emonapride and [3H]raclopride. Emonapride has a high affinity (K = 90 pM) for the dopamine D4 receptor, while raclopride has a very low affinity for this receptor (K = 240 nM); thus, any difference in the densities of these two [3H]ligands (in the absence of dopamine) could be attributed to the presence of dopamine D4 receptors. Since the binding of [3H]raclopride is sensitive to endogenous dopamine, we used Parkinson-diseased tissue which has little dopamine. We found that the densities of the two ligands were identical in Parkinson striata, indicating a low density (< 1 pmol/g) for dopamine D4 receptors in the human striatum. This low or undetectable density of dopamine D4 receptors in the striatum is consistent with other data indicating that clozapine does not have its major action in the human striatum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248850 TI - Spinal cord transection produces a long-term increase in GABAB binding in the rat substantia gelatinosa. AB - Quantitative autoradiography was used to determine changes in GABAB receptor binding in the substantia gelatinosa of the lumbar spinal cord at 4 days and at 6 weeks after a midthoracic spinal transection in rats. In the 4 day lesion animals, there was no significant change in either the density or the affinity of the GABAB binding. At 6 weeks, however, there was a 35% increase in binding density, with no significant change in affinity. The results suggest that alterations in spinal synaptic mechanisms can slowly evolve following loss of descending input to the spinal cord. PMID- 8248851 TI - Comparison of immunolocalization patterns for the synaptic vesicle proteins p65 and synapsin I in macaque monkey retina. AB - The distributions of the two synaptic vesicle proteins p65 [Matthew et al. (1981) J. Cell Biol., 91:257-269] and synapsin I [De Camilli et al. (1983) J. Cell Biol., 96:1337-1354] were compared in macaque monkey retina using pre-embedding immunocytochemistry for both light and electron microscopy. The monoclonal antibody AB-48 against p65 labeled ribbon-containing synaptic terminals of cone, rod, and bipolar cells as well as many conventional synapses of amacrine cells. In contrast, a polyclonal antiserum against synapsin I (SYN I) labeled many amacrine conventional synapses but no photoreceptor or bipolar ribbon synaptic terminals. Horizontal cell pre- and post-synaptic profiles in the outer plexiform layer were not labeled by either antibody. At the light microscopic level, the banding patterns in the inner plexiform layer also differed for the two antibodies, with four bands of AB-48 immunoreactivity in sublayers S1, S2, S4, and S5 but only three bands of SYN I immunoreactivity in S1, S3, and S5. SYN I also labeled varicose fibers in both the inner nuclear layer and the outer plexiform layer that are probably processes of dopaminergic and GABAergic interplexiform cells. Varicose fibers in the ganglion cell layer were labeled by both antibodies. These results provide the first electron microscopic immunocytochemical labeling for AB-48 and SYN I in intact retina and confirm that AB-48 labels both ribbon and conventional synaptic terminals, whereas SYN I labels only conventional synapses. PMID- 8248852 TI - Distinct kinetic binding properties of N-[3H]-methylscopolamine afford differential labeling and localization of M1, M2, and M3 muscarinic receptor subtypes in primate brain. AB - Three classes of muscarinic receptors in mammalian brain have been postulated on the basis of equilibrium and kinetic binding data. However, equilibrium binding assays alone have not permitted a clear demonstration of the localization of putative M1, M2, and M3 receptor subtypes in the brain because of the overlapping affinities of virtually all muscarinic antagonists. In the present study, the conditions for selective occupancy of the M1, M2, and M3 receptor subtypes in the brain of the rhesus monkey were based on the distinct kinetic and equilibrium binding properties of N-[3H]-methylscopolamine (NMS) at cloned m1-m4 muscarinic receptor subtypes expressed in A9L transfected cells. Quantitative autoradiography of the M1, M2, and M3 muscarinic receptor subtypes in the primate brain was performed according to the following strategy. The M1 (m1) receptor subtype was labeled directly with a non-saturating concentration of [3H] pirenzepine. The M2 (m2) subtype was labeled by incubations consisting of short, two minute pulses of [3H]-NMS after a preincubation with 0.3 microM pirenzepine to occlude m1, m3, and m4 sites. Selective occupancy of the M3 (m3) receptor (subtype) was achieved by pre-incubation with 0.5 nM unlabeled NMS to partially occlude the m1, m2, and m4 sites, equilibrium with 0.5 nM [3H]-NMS, followed by a 60 minute tracer dissociation in the presence of 1 microM atropine. In vitro autoradiography demonstrated that the M1 receptor subtype was confined to forebrain structures. M1 receptors were prevalent throughout the cerebral cortical mantle, amygdala, hippocampus, and the striatum. Low to background levels of the M1 receptor subtype were measured over the thalamus, hypothalamus, and brainstem. The M2 subtype was widely distributed with elevated densities of binding sites seen over all primary sensory cortical areas, and within discrete thalamic, hypothalamic, and brainstem nuclei. The distribution of the M3 receptor subtype was largely coincident with the pattern of the M1 sites labeled by non saturating concentrations of [3H]-pirenzepine with some notable exceptions. Within the cerebral cortical mantle, the M3 receptor exhibited an elevated gradient over the orbitofrontal gyrus and the temporal lobe. Within the striatum, the M3 subtype was elevated over the anterior and dorsal part of the caudate nucleus, while the M1 receptors were most prevalent over the ventromedial sector. Selective labeling of M3 receptors was seen over the medial division of the globus pallidus and within the substantia nigra pars reticulata. In contrast to the pattern of the M1 receptor subtype, M3 receptors were prevalent also over midline nuclei of the hypothalamus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8248853 TI - Electrophysiological evidence for a large receptor reserve for inhibition of dorsal raphe neuronal firing by 5-HT1A agonists. AB - Previous studies [Meller et al. (1990) Mol. Pharmacol., 37:231-237] have shown that a large receptor reserve exists for the inhibition of serotonin synthesis in rat cortex and hippocampus by the 5-HT1A agonist 8-hydroxy-2(di-n propylamino)tetralin (8-OH-DPAT), whereas little or no reserve exists for the lower efficacy agonists ipsapirone and BMY 7378. The current studies were undertaken to determine if the above drugs exhibit similar relative efficacies and receptor reserves in an electrophysiological model of 5-HT1A receptor activation, i.e., the inhibition of dorsal raphe cell firing. Intravenous dose response curves were constructed in untreated control rats, or in rats which received an injection of the irreversible receptor inactivator N-ethoxycarbonyl-2 ethoxy-1,2-dihydroquinoline (EEDQ, 6 mg/kg, s.c.) 24 hours before recording. All three drugs fully inhibited dorsal raphe cell firing in control rats (ED50's: 1.5 micrograms/kg, 8-OH-DPAT; 30.0 micrograms/kg, ipsapirone; 17.5 micrograms/kg, BMY 7378). However, unlike effects on serotonin synthesis, EEDQ treatments caused no depression of the maximal inhibitory response for any of the agonists, although all dose-response curves were shifted to the right (ED50's: 10.1 micrograms/kg, 6.7-fold shift, 8-OH-DPAT; 139.9 micrograms/kg, 4.7-fold shift, ipsapirone; 53.8 micrograms/kg, 3.1-fold shift, BMY 7378). Although the order of agonist efficacies was similar for both inhibition of serotonin synthesis and dorsal raphe cell firing (8-OH-DPAT > ipsapirone > BMY 7378), a large (> 50%) receptor reserve was estimated for all three drugs in this electrophysiological system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248854 TI - Chronic fluoxetine treatment upregulates 5-HT uptake sites and 5-HT2 receptors in rat brain: an autoradiographic study. AB - This study was undertaken to investigate the effect of chronic treatment with fluoxetine, a selective serotonin uptake inhibitor used widely in the treatment of depression, on the distribution and density of 5-HT uptake sites, 5-HT2 receptors, and vesicular amine uptake sites in rat brain. Fluoxetine (10 mg/kg i.p.) was administered daily for 21 days. The density of 5-HT uptake sites labelled by [3H]paroxetine, 5-HT2 receptors labelled by [3H]ketanserin in presence of tetrabenazine and vesicular amine uptake sites labelled by [3H]ketanserin in the presence of mianserin were measured by quantitative autoradiography in 22 areas of rat brain, using coronal tissue sections. Chronic administration of fluoxetine produced significant increases in the density of 5 HT uptake sites in layers of frontoparietal cortex (by 32-43%), of striate cortex (by 55%), in CA1 field of hippocampus (by 111%) and in superior colliculus (by 20%). Fluoxetine treatment also resulted in upregulation of 5-HT2 receptors in layers of frontoparietal cortex (31-38%) and in CA2-3 fields of hippocampus (by 39%). The density of tetrabenazine-sensitive vesicular amine uptake sites in the caudate-putamen was also significantly increased (by 66%). The observed alterations in 5-HT uptake site and 5-HT2 receptor densities are likely a part of adaptive neuronal changes that occur after chronic administration of fluoxetine and may be related to the antidepressant effect of the drug. PMID- 8248855 TI - Gender disparity in the academic pipeline: women in neuroscience. AB - The percentages of females at critical steps in the academic career ladder were examined for one specific field, neuroscience. There was a slight attrition among females completing the Ph.D. However, the major drop-off occurred when qualified post-doctorates did not apply for faculty positions. Remedial actions should focus on not only the graduate school environment but also on the more critical postdoctoral experience. PMID- 8248856 TI - Effects of cadmium on reproduction, an epizootologic study. AB - The Dutch part of Kempenland has been exposed to cadmium pollution since the last century. Experimental data suggest effects by cadmium on reproduction such as diminished fertility, decreased foetal growth, and specific malformations. Use was made of a historical cohort of dairy cows, ascertained from a surveillance program conducted by the regional Veterinary Health Service between 1976 and 1986. Ten dairy farms in the cadmium region were matched by size with 40 dairy farms from a reference area. Logistic models were used to calculate Odds (OR) and Rate Ratios (RR) and their 95% confidence interval. In total 4,039 exposed and 15,552 reference gestations were compared. The reasons for the slaughter of 574 exposed and of 2,824 reference cows were ascertained. A lower twinning rate [OR = 0.63 (0.47-0.84)] was found and more birth complications, for both calves [OR = 1.50 (1.25-1.80)] and cows [OR = 1.49 (1.24-1.79)]. More inseminations [OR = 1.20 (1.01-1.43)] were needed for conception in the exposed area. Deaths among twins [OR = 1.66 (0.90-3.07)] were not significantly higher. Perinatal death, premature death, age at, or reasons for, slaughter were not consistently different. These results are consistent with those of other observational and experimental studies. Extrapolation from these large domestic animals to humans is problematic, as is the more frequent extrapolation from rodents to humans. It can be concluded that longterm exposure to low levels of cadmium is associated with impaired reproduction in dairy cows. PMID- 8248857 TI - Retinoic acid induces changes in the rhombencephalic neural crest cells migration and extracellular matrix composition in chick embryos. AB - Chick embryos at 9-10 stages (Hamburger and Hamilton: J Morphol 88:49-82, 1951) have been treated with all-trans retinoid acid (RA) (0.5 microgram, 1.5 micrograms, and 2.5 micrograms) to determine the pattern and mechanism of RA induced effects on early cephalic development. We found that while 0.5 microgram RA did not produce any significant dysmorphogenesis, 2.5 micrograms RA elicited wide malformation of both cephalic and trunk regions. However, 1.5 micrograms RA produced selective and specific changes at the cephalic level, which consisted of morphological alterations, changes in neural crest cells (NCC) migration and extracellular matrix (ECM) composition. Morphological alterations included hypoplasia of the first three branchial arches, swelling of either anterior cardinal veins or dorsal aortae, and atrophy of branchial arch arteries. Concurrently NCC did not migrate away, remaining clustered on the dorsal surface of the rhombencephalon, and in some cases they shifted into the neural tube cavity. Accordingly, the second branchial arch showed a reduction of the mesenchymal cellular population. The extracellular matrix in RA-injected embryos showed changes in glycosaminoglycans (GAGs) concentration as compared with controls, that is, an increase in the non-sulphated GAGs, stained with alcian blue 8GX at 2.5 pH, and a decrease in the sulphated GAGs stained with alcian blue 8GX at 1 pH. These quantitative changes reflected alterations in the pattern of distribution and composition of the GAGs within the cephalic ECM, which specifically consisted in an increase of the hyaluronic acid and a decrease of the chondroitin sulphate. Our findings indicate that RA is involved in abnormal cephalic development, suggesting that RA may effect neural crest cell migration via changes in the GAGs of the ECM. PMID- 8248858 TI - Teratogenesis associated with oxydemeton-methyl in the stage 12 chick embryo. AB - The teratogenic potential of oxydemeton-methyl (ODM) was investigated using a stage specific localized topical method of application to the stage 12 chick embryo. A dose of 0.01, 0.05, 0.10, 0.50, 1.0, or 2.0 mg/embryo was applied to the vitelline membrane directly above the stage 12 embryo. The embryo was then returned to the incubator and monitored daily until stage 41 (15 day). At stage 41 the embryo was autopsied and examined for gross external and internal malformations, wet weight, and crown-rump length. Experimental data was compared to unopened and saline treated controls. At doses less than 0.50 mg/embryo, survival rates were high (> 80%) but when that dose was exceeded, the survival rate fell significantly (P < 0.001). A dose-dependent increase in malformation rate was seen in all treatment groups with 0.10 mg/embryo, producing a maximum malformation rate (19/33) with minimum mortality (3/36). Crown-rump lengths and wet weights were significantly less than controls in all treatment groups (P < 0.001). Anomalies were primarily seen in the musculoskeletal (ventral midline, limb, and neck) and cardiovascular (ventricular septum and aortic arches) systems. Thoracogastroschisis and ventricular septal defects were the most common combination of malformations. Our data suggest that ODM is teratogenic when topically applied to the stage 12 chick embryo. PMID- 8248859 TI - Chloroquine embryotoxicity in the postimplantation rat conceptus in vitro. AB - The embryotoxicity of the antimalarial drug chloroquine (CQ) was evaluated in vitro using the rat whole embryo culture system. CQ was found to be embryotoxic and dysmorphogenic when added directly to the culture media containing gestational day (GD) 10 rat conceptuses. Twenty-six-hr exposure to CQ elicited dose-related decreases in embryonic crown-rump length, protein and DNA contents and increases in the incidence of morphologically abnormal embryos. At 30 microM CQ, embryonic protein content was decreased to 67% and DNA content to 58% of control while the incidence of morphological abnormalities rose to 100%. Abnormal axial rotation, micro-ophthalmia, and selective cephalic hypoplasia were the most common developmental abnormalities observed. Visceral yolk sac (VYS) vasculature and blood pigmentation were also decreased in a dose-dependent manner, as was VYS DNA content (80% of control at 30 microM). VYS protein content, however, showed an alternate pattern of response, decreasing to 87% of control at 10 microM CQ but increasing to 125% of control at 30 microM. Histologic evaluation revealed that the cytoplasm of the VYS endoderm epithelium was distended due to vacuolization produced by CQ exposure. In the embryo proper, CQ inhibited cranial neural tube development and altered the morphology of cranial neural crest cells. These observations document the in vitro embryotoxicity of CQ and suggest altered VYS histiotrophic nutrition as well as direct embryonic effects as possible mechanisms of CQ embryotoxicity. PMID- 8248860 TI - Reporting of malformations in routine health registers. AB - The utility of three national registers--the malformation, birth, and hospital in patient discharge registers--in identification of malformations among 60,255 children born in 1987 in Finland was compared. Information in the malformation register is collected by specific reporting of physicians; information on the malformed children in the other two registers in 1987 through 1990 was routinely collected and identified by ICD 9-codes (740-759, 7886A). From October 1990 on, the ICD-9 codes were omitted from the birth register and malformation data were asked only in a yes-no question. In 1987, the malformation register included 1,032, the birth register 3,084 and the in-patient register 2,003 malformed infants identified up to the age of 1 year. There was underrepresentation of almost all malformation diagnoses in the malformation register. Individual linkage of the three registers showed that the malformation register revealed very few cases not recorded in the birth and the in-patient registers. With the adoption of the new recording method in the birth register beginning in October 1990, reports of malformation declined by 75%. Our analyses showed that data sources based on diagnoses collected routinely and not requiring an explicit decision on whether or not a problem is a malformation, can be more useful for routine surveillance of occurrence of malformations. PMID- 8248861 TI - Evaluation of the effectiveness of a teratogen information service: a survey of patient and professional satisfaction. AB - We have evaluated the effectiveness of the Wisconsin Teratogen Project (WTP), a teratogen information service, using two retrospective surveys. We surveyed medical professionals who utilized the WTP and patients who had received teratogen counseling in a clinic setting. The results from the medical professional survey indicated that medical professionals rely on the WTP for accurate and convenient teratogen information. A tendency to rely on the Physician's Desk Reference as an alternative resource was noted, reinforcing the need to encourage professionals to use teratogen information service projects. Major findings and conclusions from the patient survey included: patients found counseling to be most helpful if, whenever possible, numeric risks were provided; recommendations made in the absence of known risk were not considered by patients to be helpful for providing reassurance; overall compliance with recommendations was high but behavioral recommendations seemed to reduce reassurance and increase dissatisfaction with the service for some patients; and pregnant women exposed to low-dose ionizing radiation were more likely to have considered termination of pregnancy prior to referral but were not more likely to terminate following counseling. Our findings suggest that although the WTP is considered an effective service overall, teratogen counselors should give special consideration to patient and counseling characteristics as these can significantly influence the perception of the risk counseling provided. Particular attention should be given to patients with low-dose ionizing radiation exposure, since misperception of its teratogenic risk is extremely common. PMID- 8248862 TI - Fetal development in the rat following disruption of maternal renal function during pregnancy. AB - Pregnant Sprague-Dawley rats received subcutaneous injections of mercuric chloride (1-4 mg/kg) on either gestation day 7, 9, 11, or 13 to determine effects of altered maternal renal function on embryonic and fetal development. Maternal renal function, assessed by urinalysis, was markedly disrupted for at least 48 hours after treatment and resulted in decreased maternal body weight gain. Residual effects on maternal kidney weight were evident on GD 21 when the females were killed and the fetuses removed and examined for visceral and skeletal development. We did not observe an increased incidence of malformations in the offspring for exposure on any day of gestation. Maternal exposure to mercuric chloride slightly impaired fetal growth over several gestational exposures periods and changed the pattern of rib formation when exposure occurred early in organogenesis. The extent of the changes could not, however, be related to the immediate degree or duration of altered maternal renal function. Rather, we found correlations between lasting effects of exposure as measured by maternal renal weight on GD 21 and super-numerary lumbar rib induction on GD 7 and 9; while for the relationship with fetal weight, the strongest correlation with maternal kidney weight occurred following exposure on GD 9 (P < 0.01), with weaker correlations (P < 0.10) for GD 7 and 13 exposures. Maternal serum urea was negatively correlated with fetal weight from the GD 7 exposure. This study supports the concept that some specific forms of maternal toxicity may be associated with limited manifestations of developmental toxicity, but, in general, embryonic development was observed to proceed normally in the presence of marked disruptions in maternal renal physiology. Maternal toxicity, especially as routinely measured in developmental toxicity studies, cannot be looked upon as a single disease. Instead, each manifestation must be examined for what it signifies to the physiology and well being of the female and for cause-and-effect relationships with fetal observations. Maternal toxicity and developmental toxicity should not be etiologically linked solely because of their concurrent appearance on the dose-response curve. PMID- 8248863 TI - Relationship between abnormal somite development and axial skeletal defects in rats following heat exposure. AB - The effects of in vivo heat exposure on gestation day (GD) 10 rat embryos were evaluated on GD 11 to determine the relationships between morphological sequelae following in vivo and in vitro exposures and between effects detected on GD 11 and those observed in postnatal day (PND) 3 pups. Anesthetized rats were exposed to 42 degrees C in a warm air incubator until their rectal temperatures reached 41 degrees C or until a rectal temperature of 42-42.5 degrees C had been maintained for 5 minutes. Heat-exposed embryos exhibited a significant decrease in growth parameters including head length, somite number, and protein content/embryo versus controls. These changes correlated well with in vitro effects from an earlier study (G.L. Kimmel et al., '93). Among the morphological endpoints which were slightly delayed in development were the caudal neural tube, branchial bars, forelimb and hindlimb. The only effect on the embryos that could not be explained as a transient delay in development induced by heat was the induction of unsegmented somites. Additional embryos were exposed to 42 degrees C for 15-20 min in vitro and examined specifically for unsegmented somites, which were observed in 47% of embryos exposed to 42 degrees C in vivo or in vitro. This phenomenon was observed in somites 9-20, i.e., those that give rise to cervical and thoracic vertebrae and ribs. These results correlated well with the axial skeletal malformations observed in PND 3 pups exposed to the same heat treatment (C.A. Kimmel et al., '93). PMID- 8248865 TI - What do patients think about their physicians? PMID- 8248864 TI - Glucose and scyllo-inositol impair phosphoinositide hydrolysis in the 10.5-day cultured rat conceptus: a role in dysmorphogenesis? AB - Culture of the postimplantation rat conceptus from gestational day 9.5-10.5 in media supplemented with d-glucose or scyllo-inositol decreases tissue myo inositol and phosphoinositides with a concomitant increase in dysmorphogenesis. A number of mitogenic agents initiate cellular proliferation and differentiation through receptors coupled to phosphoinositide hydrolysis. To test whether the decrease in conceptus phosphoinositides is associated with a reduced phosphoinositide hydrolytic response, we developed a protocol to stimulate phosphoinositide hydrolysis. Phosphoinositide hydrolysis was monitored by measurement of [3H]inositol phosphates after preincubation in serum free media. We examined the ability of serum, platelet-derived growth factor (PDGF), epidermal-derived growth factor (EGF), insulin-like growth factor 1 (IGF-1), insulin-like growth factor 2 (IGF-2), endothelin-1 (ET-1), and endothelin-2 (ET 2), to stimulate phosphoinositide hydrolysis. As measured by [3H]inositol monophosphate ([3H]InsP1) accumulation, normal rat seru, ET-1, and ET-2 stimulated phosphoinositide hydrolysis 47%, 420%, and 154% above the basal rate observed in serum free controls. EGF stimulated a statistically insignificant 15% increase while PDGF, IGF-1, or IGF-2 were without effect. We further characterized ET-1 stimulated phosphoinositide hydrolysis. Dose-response studies disclosed that incremental increases in [3H]InsP1 (129-420%) are observed over a concentration range of 10-1,000 nM. Maximal stimulation was not reached even at 1,000 nM. Temporally [3H]InsP1 and [3H]InsP3 levels increased linearly during incubation periods of 15-60 min. We further analyzed ET-1 stimulated phosphoinositide hydrolysis in 10.5-day conceptuses cultured for 24 hr in media containing high concentrations of glucose (23.3-56.6 mM) or scyllo-inositol (0.55, 5.5 mM). Under these dysmorphogenic conditions that concomitantly decrease the phosphoinositide precursor pool the response to ET-1 was blunted 28-76% for glucose and 29-65% for scyllo-inositol. This suggests that the effect of glucose and scyllo-inositol on lowering phosphoinositide precursor pools also results in a decrease in the response to agonists using the inositol/lipid intracellular pathway. This impaired signaling response may contribute to initiating dysmorphogenic events in diabetic embryopathy. PMID- 8248866 TI - Physicians reaching out to their communities through CMS programs. PMID- 8248867 TI - Physicians find good, bad news in Clinton budget. PMID- 8248868 TI - Troubling waters: sister cities struggle with health conditions on the US-Mexico border. PMID- 8248869 TI - TMA targets private insurers to cut red tape, hassles for physicians. PMID- 8248870 TI - Texas physicians on the nursing home front. PMID- 8248871 TI - Dedication and a system that makes sense. PMID- 8248872 TI - A return to old-fashioned doctoring. PMID- 8248874 TI - The good, the bad, and the underfunded. PMID- 8248873 TI - Making differences in small ways. PMID- 8248875 TI - Bridging the age gap. PMID- 8248876 TI - Studies target ethnic and racial differences in drug metabolism. PMID- 8248877 TI - Breast implant controversy: lawyers say physicians not to blame. PMID- 8248878 TI - New option in forming group practices. PMID- 8248879 TI - Congestive heart failure. AB - Congestive heart failure is important to the physician who treats elderly patients because of its incidence, prevalence, and high morbidity and mortality and because it often necessitates repeated hospitalizations. Presentation may be atypical and the condition may be precipitated by many of the ills that occur routinely in geriatric practice. Diastolic dysfunction may complicate diagnostic evaluation. Many of the symptoms respond gratifyingly to therapy. Congestive heart failure ranks among the most treatable of the geriatric syndromes. The physician who cares for older patients should be constantly alert for its presence and should keep abreast of the current therapies available and acquire experience in their use. PMID- 8248880 TI - Hispanic elders in Texas: implications for health care. AB - Hispanic elders are a large, rapidly growing subgroup of the Hispanic population in Texas. This population continues to suffer from a high incidence of non insulin-dependent diabetes mellitus, with similar or lower rates of cardiovascular disease than non-Hispanic whites have. Barriers that may modify potentially effective diagnostic or appropriate treatment plans include communication problems, access issues, and use of informal health-care services. Knowledge and attention to these issues may improve compliance and decrease morbidity of this cohort. This article summarizes important information affecting the health of Hispanic elders in Texas and alerts practitioners to issues that may affect the care of their older Hispanic patients. PMID- 8248881 TI - Challenges to quality care in Texas nursing facilities. AB - The American population is aging. Predictions for the future estimate that nearly 50% of those over age 65 will need long-term institutionalized care in nursing facilities at some time. Texas has gained national attention for the difficulties it faces in providing quality care to this vulnerable group. Nursing facilities face challenges in reimbursement, regulation, medical direction and nursing care, quality improvement, and ethical dilemmas. This article will attempt to present these challenges and offer solutions consistent with the goals and desires of consumers and providers. PMID- 8248882 TI - Osteoporosis: considerations in evaluating and managing the older patient. AB - Osteoporosis is a common disorder that results in 1.3 million fractures each year in the United States. The resultant morbidity and mortality are substantial. While there has been considerable progress in our understanding of this disease, research studies frequently include only early postmenopausal individuals. At the same time, a number of important pathophysiologic and clinical differences exist between early postmenopausal and elderly individuals. These considerations raise the issue of the appropriateness of generalizing evaluation and treatment recommendations from younger to older patients. This review emphasizes factors important in the assessment and treatment of established osteoporosis in older patients. PMID- 8248883 TI - Where were the DOs? PMID- 8248884 TI - Office management of late-life depression. AB - Depression is often misunderstood and underappreciated as a genuine clinical entity throughout the life cycle. In elderly persons especially, it can be underdiagnosed and undertreated. Office physicians are the most important resource for early intervention in late-life depression where results can be as successful as with younger age groups. Physician awareness of the normal biopsychosocial changes of late life is essential to skillful assessment and intervention since somatic disorders and their symptoms frequently overlap those of psychological origin. Tricyclic antidepressant drugs remain the core of modern medical intervention, but their appropriate use in elderly patients requires special care and finesse on the part of office-based physicians. PMID- 8248885 TI - A stochastic model for genetic linkage equilibrium. AB - Linkage equilibrium is an independence condition among the alleles at a set of gene loci. Equilibrium or disequilibrium only makes sense relative to some reference population of a species. If the loci all occur on the same chromosome, then linkage equilibrium holds provided a random representative of that chromosome from the reference population displays independent alleles at the various loci of the set. Classical deterministic population genetics theory shows that linkage equilibrium is approached asymptotically after many generations of random mating in a reference population of infinite size. The current paper considers a Markov chain model for the establishment of linkage equilibrium in a population of finite size. The states of this Markov chain correspond to counts of chromosomes of various types. Because the chain is reversible, the equilibrium distribution can be explicitly computed. Partial characterization of the geometric rate of convergence of the chain to equilibrium is possible using a strong stationary stopping time. PMID- 8248886 TI - Filling a gap in the prediction of the equilibrium genetic variance. AB - We derive analytical predictions for the variance and kurtosis of the equilibrium distribution of allelic effects under stabilizing selection and drift, and for any value of the kurtosis of the distribution of the mutational effects. Numerical results relative to the equilibrium genetic variance are compared with other analytical predictions and with simulation results from the literature. Our prediction is superior for relatively weak selection (or relatively low variance of the mutational effects) and not to small population sizes, where previous methods overestimate the equilibrium genetic variance. The behavior of the equilibrium kurtosis is also illustrated. PMID- 8248887 TI - Recombination modification with X-linked characters. PMID- 8248888 TI - The evolution of interference: reduction of recombination among three loci. AB - Crossover events along chromosomes do not occur independently, but influence the probability of other nearby events. The most common interaction between nearby crossover events is inhibitory: a crossover event tends to reduce the probability of other such events nearby, and this is called positive interference. A crossover event may increase the probability of events nearby, and this rare phenomenon is called negative interference. In this paper, we use numerical methods to investigate how interference among three loci would evolve if it were under the genetic control of a fourth, selectively neutral locus. We first discuss the effect of interference on the overall rate of recombination among the three loci, and then show that, under a variety of conditions, interference evolves in the same way as would be predicted based upon its effect on the overall rate of recombination. That is, the overall rate evolves in the same direction as would the rate at a locus that controls recombination between two loci directly. We then check for the existence of viability-analogous Hardy Weinberg equilibria in the four-locus model of interference modification. PMID- 8248889 TI - Typing of serum-soluble HLA-B27 antigen by ELISA. AB - An ELISA using serum as soluble HLA antigen source was developed for HLA-B27 typing. Two sandwich assays were run in parallel. The first assay utilized a monoclonal antibody (mAb) reacting with a determinant expressed by both HLA-B7 and B27 antigens; the other assay utilized a mAb reactive with HLA-B7 antigens but not with HLA-B27 antigens. After incubation with serum samples, bound HLA antigen was detected using an anti-beta 2m antibody conjugated to peroxidase and a chromogenic substrate. Absorbance of each well was measured at 490 nm. Based on analysis of absorbances obtained with panels of specimens of known HLA phenotypes, a mathematical algorithm was developed to derive the specimen HLA-B27 phenotype from its ELISA absorbance values. Despite the lack of monospecific mAb, an accurate HLA-B27 typing was possible. 362 specimens (including 151 HLA-B27 positive) were tested. Agreement between microlymphocytotoxicity and ELISA was 99.2%. No correlation between the level of HLA-B27 antigen reactivity and the amount of total HLA class I antigen in serum was observed. This report demonstrates the possibility of using serum-soluble HLA antigen and ELISA technology for histocompatibility testing. The assay offers several significant advantages over microlymphocytotoxicity: no need for cell preparation, batch testing capabilities and objective, reproducible interpretation of results. PMID- 8248890 TI - Biochemical and functional characteristics of soluble MHC molecules derived from H-2Ld/Q10d chimeric gene. AB - We have constructed a chimeric class I gene in which the 5' half of the H-2Ld gene is linked to the 3' half of Q10d. The resulting H-2Ld/Q10d protein is homologous to the native H-2Ld heavy chain for the three external domains except for an Arg to His substitution at position 260. The transmembrane and intracytoplasmic domains of the H-2Ld chain are replaced by the short low hydrophobic transmembrane-like domain of the Q10d chain. Following DNA-mediated gene transfer into mouse L cells, transformants were selected for the presence of specific mRNA. Radiolabelling and immunoprecipitation analysis revealed secretion of a 48-46 kd chain weakly associated with beta 2-microglobulin. This molecule reacts with H-2Ld-specific mAb that identify determinants on the first and second domains as well as with an anti-Q10 carboxyl-terminal peptide antiserum, but is not recognized by a mAb specific for a determinant of H-2Ld third domain. The integrity of antibody reactivity of the first and second domains together with beta 2-microglobulin association suggest that our molecule may be considered a good soluble counterpart of the native membrane H-2Ld molecule with which to perform functional studies. In order to analyze the immunogenic capacities and T cell recognition of the soluble H-2Ld molecules, T-cell lines were produced from mice of various inbred strains immunized with supernatant from H-2Ld/Q10d transfected fibroblasts. Characterization of these T cells revealed that they expressed a CD4+CD8- phenotype, and recognized H-2Ld/Q10d products in a class II restricted manner. PMID- 8248891 TI - Soluble MHC class I antigens (sHLA) and anti-HLA antibodies in heart and kidney allograft recipients. AB - Soluble HLA (sHLA) antigens were measured in a panel of 50 renal and 50 cardiac graft recipients by a quantitative ELISA over periods ranging between 2 months and 3 years. 72% of the renal patients and 68% of the cardiac graft patients experienced episodes of acute rejection during the observation period. sHLA were elevated to over 2-5 times the normal levels up to 10 days before histological evidence of rejection. This duration was for each individual patient very variable. Cytomegalovirus (CMV) infections did not appear to elevate sHLA levels. Donor-specific HLA-A2 was measured in only 1 cardiac recipient. 15% of the sera obtained from cardiac patients had cytotoxic anti-HLA antibodies, when tested against a panel of 50 test cells. After thermal inactivation at 56 degrees C, 42.5% of the sera became positive against at least 10% of the test cells. This finding suggests thermal dissociation of immune complexed anti-HLA antibodies. sHLA measurement and careful monitoring of serum for the presence of cytotoxic anti-HLA antibodies in graft patients are useful techniques which could help predict the long-term fate of an allograft. PMID- 8248892 TI - Human interleukin-2 and lymphoproliferative (T-helper cell) responses to soluble HLA class I antigens in vitro: I. Specificity for polymorphic domains. PMID- 8248893 TI - A single amino acid substitution at residue 167 forms a novel HLA-B51 subtype. PMID- 8248894 TI - A novel HLA-DRB1*01 allele (DRB1*0104). PMID- 8248896 TI - [Thoughts on the reform of veterinary studies]. PMID- 8248895 TI - Increased serum HLA class I molecule levels in elderly human responders to influenza vaccination. AB - Increased cell surface expression and serum levels of HLA class I molecules have been shown to occur in normal elderly humans. Elderly subjects (65 years and older) generally also have a poorer response to influenza vaccine than younger adults. Some 30 to 40% of elderly subjects do not respond to influenza vaccination. Sera from 20 elderly subjects with a positive response to influenza vaccine (HAI antibody titer > 40) and 20 subjects who did not respond (HAI antibody titer < 40) were randomly selected and the levels of soluble HLA class I molecules determined before and after vaccination. The subjects who responded had higher serum HLA class I levels both before and after vaccination than did non responders. Two weeks after vaccination non-responders showed similar serum HLA class I levels (1.78 mg/ml +/- 0.62) as compared to prevaccination levels (1.73 mg/ml +/- 0.49). The responders showed significantly higher levels of soluble HLA class I molecules following vaccination (2.64 mg/ml +/- 0.99) as compared to pre vaccination (2.08 mg/ml +/- 0.85, p = 0.00001). Our results suggest that measuring soluble HLA class I molecules in the serum may be useful in assessing the adequacy of response to influenza vaccination in the elderly population. PMID- 8248897 TI - [Health and disease in humans and animals over the course of history]. AB - A historical review of the terms health and sickness in human beings and animals. The terms health and sickness have changed from ancient history to the present day and have also been modified. The notions of what life really is, when it starts and when it ends, have contributed to these changes. Historically, health and sickness have generally been interpreted anthropocentrically in the course of time, rarely has the companionship between man and animal been taken into account. A closer look over the past three millennia as to the science of the dynamics of diseases, the humoral pathology and the solidistic pathology, leads to the conclusion that the terms sickness and health have only been understood in general, not in detail. In the last decades of the 19th century and up to the present day an essential change of the notion of sickness was initiated through cellular pathology by R. Virchow (1821-1902). Eventually, bacteriology, virology, molecular biology and gene technology have established new standards with regard to health and sickness and have paved the way to the present holistic medicine, which is based on science. Sickness is defined as a verifiable divergence from the norm of anatomical, physiological, immunological and psychic conditions of an organism and, accordingly, as a pathological form of being, implying a disturbance of homeostasis. The legal issues such as the federal laws on epidemic diseases of human beings and animals and on insurance are dealt with in conclusion. PMID- 8248898 TI - [The clinical case. Necrosis of the ends of both hindlimbs of a calf]. PMID- 8248899 TI - [The effects of glucocorticosteroids on lipolysis stimulated by fasting and liver function in cows]. AB - During a five-day period prednisolone (200 mg/d) reduced the rise of FFA, beta hydroxybutyrate and bilirubin concentrations in the blood plasma of three cows compared with three control animals. The liver fat content remained within the physiological range. The reduced lipolysis in the experimental group was the result of a reactive hyperinsulinaemia, a more effective energy metabolism and a diminished metabolic activity. Harmful effects on the liver function could not be observed. Plasma cortisol concentration was reduced for only a short time. PMID- 8248900 TI - [Clinicochemical blood values in newborn calves. 2. Repeated studies in the same animals]. AB - Blood samples taken by venipuncture in calves within 12 hours after delivery and again in the same animals within 5 to 23 days of postnatal life were analyzed for hematocrit and fifteen other biochemical values. The changes of blood values between these two sampling points in each individual calf were investigated by regression analysis. Remarkable individual reactions and effects of the constitution of the animal immediately after delivery on the level and on the changes of the biochemical values could be observed as well as great deviations of some blood values of twin calves too. The scattering of some blood values and their changes in the first days of postnatal life are discussed regarding the maturity of the calves as well as the possible incidence of deficiency states or the setting of a new level for these physiological values. PMID- 8248901 TI - [Case reports of hemorrhagic diathesis in calves with bovine diarrhea virus infection]. AB - Clinical and pathological findings in six veal calves suffering from haemorrhagic diathesis are reported. Further pathological and virological results were highly suggestive of mucosal disease (BVD). No virus isolation or classification was possible. The post mortem results indicated that the virus might have been cytopathogenic. The cases are discussed and compared with similar field and experimental publications dealing with thrombocytopenia in veal calves in the USA. These are the first reports of cases published in Germany. PMID- 8248902 TI - [Effects of different methods of tooth resection in suckling piglets]. AB - Epizootic examinations were made on 796 newborn piglets to test the consequences of the resection of a piglet's canine and lateral incisor teeth on its first day of life. As a comparative examination the resection was carried out with side cutter pliers or with the teeth grinder PIGMATIC 110. A third group of piglets was not treated. Histological, radiographic and bacteriological examinations were made of 10 piglets--in each case with piglets with the resection with side-cutter pliers or with the teeth grinder. 48% of ground teeth reacted with an inflammation of the pulp. The clipping of the crown of teeth with side-cutter pliers caused pulpitis in 92% of the teeth. Inflammation of the gingiva could almost only be seen around clipped teeth. Splinters only occurred with teeth which had been clipped with side-cutter pliers. Gingivitis and pulpitis extended along the splinters to the bottom of the root. Bites among the litter mates were more frequent in the group of piglets whose teeth had not been treated than in the group of piglets with resected teeth. Compared to the grinding or the clipping of needle teeth, bites to the sow's udder were much more frequent if the resection had been omitted. During the whole period of examination the mortality of piglets was at its lowest in the group of piglets with clipped teeth. The development of grinding instruments for the resection of needle teeth presents a method which reduces the negative consequences of the conventional resection with side-cutter pliers and shows clearly the advantages of resection. In terms of animal welfare, teeth resection is an amputation and therefore every case requires veterinary justification. PMID- 8248903 TI - [Naturally-acquired microsporidia (Encephalitozoon cuniculi) infections in hens]. AB - During a short period of time, four approximately ten months old chickens from an egg production (group A) unit developed symptoms of apathy, lameness, mild diarrhoea and loss of weight. The layers were killed and submitted for necropsy. For the first time, microsporidia infection, probably Encephalitozoon cuniculi (E.c.) was diagnosed in chickens. The diagnosis was mainly based on immunohistochemical methods. E.c. was found particularly in the cells of the cutaneous mucous membrane of the crop and in epithelial cells, the cells of the propria mucosae and the smooth muscle cells of the small intestine. E.c. also frequently occurred in the fibres of the skeletal muscle. Intracellularly, E.c. and its development stages were located primarily in the cytoplasm and less frequently in the nucleus of cells. Retrospective investigations on tissue samples from a domestic cock and 9 hens by immunohistochemical methods also revealed occurrence of this microsporidian. The results showed that in chickens E.c. infections can cause multi-symptomatic illness but may also be clinically inapparent. PMID- 8248904 TI - [Experiences with long-term intravenous therapy using teflon catheters in 80 horses]. AB - In a retrospective study the tolerance to Teflon coated catheters in long-term intravenous medication was evaluated in 80 horses. Catheters were inserted into the jugular vein and remained there for 3 to 30 days (average 8.6 days). Catheters were flushed using an heparinized solution after each medication administration. The site of catheter placement was evaluated daily for swelling, pain and venous distensibility respectively. Swelling at the site of insertion was noted in 10 horses, a small subcutaneous abscess formation was identified in one horse. Fourteen sonographic evaluations were performed on 10 horses, demonstrating perivascular changes in four. Thrombus formation did not occur. In all horses the catheter could be identified floating freely within the vascular lumen. Catheter failure occurred in nearly a third of all cases either as a result of mechanical failure or of pull-out by the horse. After removal of the catheter no adverse side effects attributable to the mode of medication were noted in any of the horses. In conclusion, intravenous medication using Teflon coated catheters is a suitable alternative to other modes of administration. PMID- 8248905 TI - [Recent information about the etiopathogenesis of paretic-paralytic forms of herpesvirus infection in horses]. AB - From spring 1990 to summer 1991 we investigated 21 horses with clinical symptoms of EHV-infection by means of serological and virological methods including DNA hybridization to identify the causative agents. The results indicated that, as already reported by us, EHV4 may also cause the paralytic form of the infection. The possibility of double infection with EHV4 and EHV1 cannot be excluded. In 3 out of 21 affected horses we could investigate brain tissue and/or spinal fluid by Dotblot hybridization with EHV1 and EHV4-DNA. The investigated samples of all three horses showed hybridization with EHV4-DNA, without or with less pronounced reaction with EHV1-DNA. The results were confirmed by serological investigation. Brain tissue and cerebrospinal fluid from two horses with paretic or paralytic disorders (1979 and 1980) was also investigated by DNA hybridization. In the liquor of one horse--a 5-month-old foal with neonatal ataxia--we detected EHV1 DNA. The other horse showed a strong reaction with EHV1 and a weaker reaction with EHV4 in its brain material and no hybridisation in the cerebrospinal fluid. The results are discussed. PMID- 8248906 TI - [Muscular dystrophy in a cat]. AB - A case of muscular dystrophy in a 1-year-old male castrated Domestic Shorthair cat is presented. The most striking clinical features were regurgitation, a stiff gait, an increased muscle tone and exercise intolerance. Serum biochemistry panels showed a marked increase in the muscle specific enzyme creatine kinase, and moderately elevated levels of LDH, AST and ALT. Spontaneous electrical activity of skeletal muscles in the form of "bizarre high frequency discharges" and "myotonia-like repetitive discharges" were registered. Gross pathology revealed a marked hypertrophy of the skeletal muscles. The main histopathological changes were myofiber necrosis and calcification, variation in fiber size, hypertrophied muscle fibers of type I and type II and fiber splitting. Indirect immunofluorescence showed dystrophin deficiency. Feline muscular dystrophy resembles the X-linked human Duchenne muscular dystrophy (DMD). Besides the X linked muscular dystrophy in the mouse and Golden Retriever the feline muscular dystrophy could represent another valuable animal model for the study of DMD. PMID- 8248907 TI - [The transmucosal measurement of oxygen partial pressure (PO2) in dogs]. AB - A transmucosal measuring method for monitoring PO2 continuously is described. In an undisturbed microcirculation, the transmucosal PO2 measurement may complement the measurement of PO2 in arterialized blood in order to diagnose several respiratory diseases as well as heart diseases and circulatory disorders. Investigations with uraemic patients and with dogs under conditions of neuroleptoanalgesia serve to explain the influence of microcirculation on transmucosal PO2. This influence allows an inference from transmucosal PO2 to the function or dysfunction of microcirculation in an organism. Therefore transmucosal PO2 measurement offers the possibility of monitoring high-risk patients continuously. PMID- 8248908 TI - [Serologic diagnosis of canine leishmaniasis: the dot-ELISA as a simple alternative to conventional ELISA]. AB - Serology is an important tool for the diagnosis of canine leishmaniasis, since the direct detection of the parasite remains negative in many cases. A dot-ELISA was developed, which can be read without using a photometer. For this purpose assay strips were prepared, which were divided into a control field (staining indicates normal function of reagents) and four reaction fields. The reaction fields were coated with four different concentrations of Leishmania antigen and allowed semiquantitative readings of test results. The comparison of the reactions of 73 sera (20 healthy dogs, 24 dogs with leishmaniasis, 15 dogs with trypanosomiasis and 14 dogs with babesiosis) in the dot-ELISA with the results of a conventional microtiter plate ELISA showed a significant correlation between the two methods. PMID- 8248909 TI - [Congenital "ectromelia" in fur-bearing animals caused by Orthopoxvirus muris]. AB - Orthopox virus infection is endemic in farms with fur-bearing animals in the Czech Republic (Bohemia and Moravia). This disease is called ectromelia of silver foxes and minks. The infection is congenitally transmitted and manifests itself in reproductive disorders, stillbirth or birth of sick neonates. Adult animals are usually free of clinical symptoms. The infective agent was isolated from recent outbreaks and was identified as a mouse pox virus (Orthopoxvirus muris) by its cultural and immunological characteristics. The significance of this pox virus infection, hitherto not described in Western Europe, is discussed. PMID- 8248910 TI - Protection of BALB/c mice against methicillin-resistant Staphylococcus aureus infection by intraperitoneal administration of nucleoside-nucleotide mixture. AB - The effect of intraperitoneally administered nucleic acid components (nucleoside nucleotide mixture) on the recovery from methicillin-resistant Staphylococcus aureus (MRSA) strain 8985N infection was studied in mice. Two experiments were conducted in which BALB/c mice were fed a nucleotide-free 20% casein diet for 30 days. On the 10th day, the mice were inoculated intravenously with viable MRSA organisms. The mice were intraperitoneally administered nucleoside-nucleotide mixture or saline (control) daily from the onset of the experiment (experiment 1) or from the day of inoculation (experiment 2). The survival rate in the nucleoside-nucleotide group in experiment 1 (70%) was significantly higher than that in the saline group (20%) (p < 0.05). In experiment 2, the survival rate in the nucleoside-nucleotide group (55%) tended to be higher than that in the saline group (36%) without statistically significant difference; furthermore, in the surviving mice, the viable MRSA organisms recovered from the spleen and the kidney were fewer in the former group than in the latter group (p < 0.05). The studies showed that the intraperitoneal administration either before or after the MRSA challenge of the nucleoside-nucleotide mixture was effective for the recovery of the mice from the infection. PMID- 8248911 TI - Prognostic significance of simultaneous infiltration of HLA-DR-positive dendritic cells and tumor infiltrating lymphocytes into human esophageal carcinoma. AB - The distribution of HLA-DR-positive dendritic cells (DR+DCs) and tumor infiltrating lymphocytes (TILs) was investigated in 67 patients with squamous cell carcinoma of the esophagus. DR+DCs were chiefly present among the cancer cell nests, and few were seen in the stroma. As clusters, UCHL-1-positive T-cells (UCHL1-Ts) were detected in the tissue around the cancer cell nests and as individual cells within tumor nests. Some UCHL1-Ts in the tumor nests showed direct contact with DR+DCs. In addition, a few OPD4-positive helper/inducer T cells were found among cancer cells, and were densely present in the stroma. A correlation was demonstrated between the number of DR+DCs and UCHL1-Ts in each patient (R = 0.4547, p < 0.01). In addition, dense tumor infiltration by both DR+DCs and UCHL1-Ts within the cancer cell nests was related to a better prognosis (p < 0.01). PMID- 8248912 TI - Effect of acidosis on insulin binding and glucose uptake in isolated rat adipocytes. AB - To assess the mechanism of insulin resistance in metabolic acidosis, we investigated insulin receptor binding and glucose uptake in isolated rat epididymal adipocytes at three pHs, acidic (pH 6.8), neutral (pH 7.4) and alkaline (pH 8.0) groups. After male Sprague-Dawley rats weighing 220-240 g were submitted under chloral hydrate anesthesia, epididymal fat pads were dissected and isolated. [125I]-insulin binding rate was not different between alkaline and neutral groups, but that of acidic group was markedly decreased to approximately 30% of neutral group at maximum specific insulin binding rate. Scatchard analysis showed that numbers of insulin receptors were not different among three groups, but values of binding affinity in acidic group was significantly lower than those of the others. 2-[3H]deoxy-D-glucose uptake of alkaline and neutral groups were increased to reach steady state immediately, but that of acidic group increased slowly and reached at maximum which was lower than the other two groups. Furthermore, 2-deoxy-D-glucose uptake against amount of bound insulin was lower in acidic group than other two groups. From these results, it is suggested that insulin resistance in metabolic acidosis resulted from the combined defects in binding affinity and post-binding process in the insulin receptor. PMID- 8248913 TI - Changes of 31P-MR spectroscopy in experimental tumors following radiotherapy combined with 5-fluorouracil compound (UFT). AB - The energy metabolism of FM3A tumors in C3H mice following radiotherapy (20 Gy or 40 Gy), 5-fluorouracil compound (UFT) chemotherapy or radiotherapy (20 Gy) combined with UFT was investigated by in vivo 31P-MR spectroscopy on day 2 after treatment. The effects were compared with the growth curves of the tumors and with the cytokinetic and histological changes in the tumors on day 5 after treatment. The combined group (20 Gy+UFT) showed growth curves similar to the 40 Gy group. Cytokinetic and histological changes of the combined group were also similar to those of the 40 Gy group, suggesting supra-additive effect of UFT. 31P MR spectra of the combined group were similar to those of the 40 Gy group which were characterized by increase of intracellular pH(pHi), decrease of Pi/beta-NTP and of PME/total phosphorus-signal ratios. Spectra of the UFT group and the 20 Gy group, on the contrary, showed decrease of pHi, increase of Pi/beta-NTP and of PME/total phosphorus-signal ratios. Changes detected by 31P-MR spectroscopy on day 2 were considered to reflect early metabolic events which were correlated to the growth curve, cytokinetic and histological alterations observed on day 5 after treatment. PMID- 8248914 TI - Female systemic lupus erythematosus in Miyagi Prefecture, Japan: a case-control study of dietary and reproductive factors. AB - To investigate risk factors of systemic lupus erythematosus (SLE) in relation to diet and reproduction, a population-based case-control study was conducted during the period from October 1988 to October 1989 in Miyagi Prefecture in northeastern Japan. Included in the study were 52 female patients with the recent SLE onset. Two sex- and birth year-matched (+/- 2 years) controls for each patient were selected from the general population. The analysis on diet showed that the frequent intake of meat was associated with an increased risk (frequent vs. rare, relative risk (RR) 3.36; 95% confidence interval (CI) 1.10-10.24) and that the patients preferred fatty meat such as beef or pork. The analysis on menstrual history revealed that menstrual irregularity was also associated with an increased risk (RR 3.79; 95% CI 1.43-10.01). These results suggest that dietary and reproductive factors may be responsible for the onset and the progression of SLE. PMID- 8248915 TI - Rises in urinary bicarbonate contents and pH of adult Tokyo citizens. AB - Urinary bicarbonate has been considered to be negligible, and the pH of urine to be low (around pH 5.5) for these fifty years. It was, however, found that the contents of bicarbonate and pH of the urine of adult Tokyo citizens (435 adults including 148 males and 287 females) substantially increased. The findings may be correlated with increased inhaled amounts of carbon dioxide which finally causes the increase of urinary excretion of bicarbonate, though evidence that the gas accumulates in the atmosphere of Tokyo due to air pollution has not been directly demonstrated. PMID- 8248916 TI - Lack of direct action of atriopeptidase inhibitor on cellular pH regulation in rabbit S2 proximal straight tubules. AB - While the in vivo diuretic and antihypertensive effects of the newly-developed drug Candoxatril (UK-79300), a prodrug of a selective atriopeptidase inhibitor UK 73967 (API) are well established, the mechanism of its diuretic action is not yet fully understood due to the lack of information about its direct effects on proximal tubules. To elucidate whether this atriopeptidase inhibitor has any direct effects on proximal tubules, we examined the effect of API on intracellular pH (pHi) of the in vitro microperfused proximal S2 segment of rabbit kidney, using the pH-sensitive fluorescent dye, (2',7')-bis(carboxyethyl) (5,6)-carboxyfluorescein (BCECF). In the steady-state condition, pHi was 7.13 +/- 0.02 (n = 19). Atriopeptidase inhibitor (API) at 100 microM added to the bath changed pHi by only 0.02 +/- 0.07 unit (n = 7, p > 0.05) in 10 min. The same concentration of API in the lumen changed pHi by 0.03 +/- 0.02 unit (n = 6, p > 0.05). To test whether the synergistic effects of API on the luminal and basolateral transport systems prevented the apparent change in pHi, we also examined the effect of API in the presence of amiloride in the lumen. The inhibition of Na+/H+ antiporter by the addition of 1 mM of amiloride to the lumen caused no change in pHi response to basolateral API application. API was without effect on pHi also in the presence of atrial natriuretic polypeptide (ANP). These results suggest that API per se has no significant effect on the process of proximal acidification over a short time period. PMID- 8248917 TI - The augmented blood pressure response to chronic angiotensin II infusion in partially nephrectomized rats. AB - To assess potential roles of circulating levels of angiotensin II (Ang II) in the regulation of blood pressure in rats with reduced functional renal mass, we studied the blood pressure response to chronic infusion of Ang II at a subpressor dose in partially nephrectomized rats. Male Sprague-Dawley rats were subjected to 1/6, 2/6, 3/6, 4/6 and 5/6 nephrectomy, and sham operation a week before the infusion of Ang II at a rate of 400 micrograms/kg/day delivered intraperitoneally by osmotic minipumps for 14 days, or vehicle alone served as controls. Infusion of Ang II was associated with a slight but significant increase in systolic blood pressure in 1/6 nephrectomized rats (p < 0.05), whereas it did not induce a significant change in systolic blood pressure in sham-operated rats. Hypertension during Ang II infusion was more prominent in 2/6 nephrectomized rats than that in 1/6 nephrectomized rats (p < 0.05). However, there was no significant difference in hypertension during Ang II infusion among groups in 2/6, 3/6, 4/6 and 5/6 nephrectomized rats. Blood urea nitrogen and serum creatinine levels, endogenous creatinine clearance, urinary excretion of protein as indices of renal function, and urinary sodium excretion were not affected by chronic infusion of Ang II in 1/6, 2/6, 3/6, 4/6 and 5/6 nephrectomized, and sham-operated rats. There was also no significant difference in circulating levels of Ang II among Ang II infused groups in 1/6 and 2/6 nephrectomized rats. These results indicate that elevated levels of circulating Ang II are capable of causing a sustained hypertension in rats with reduced functional renal mass.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8248918 TI - Cephalometric study on the influence of vertical traction of teeth on maxillofacial bones in young dogs. AB - The influence of one-sided vertical traction of teeth on maxillofacial bones was investigated using cephalometric radiographs and tetracycline-labeling in young dogs. On the frontal view of the dry skull, traction resulted in a downward distortional deformity of the maxilla including the nasal floor. Cephalometry of radiographs revealed a downward displacement of the upper premolar and incisor teeth, medial maxillary region, and alveolar process of molar, canine and incisor on the right side, where traction was applied. Displacement was from 2.2 +/- 0.4 (mean +/- S.D.) mm at P3 to 0.8 +/- 0.2 mm at I1. In the control group without traction, it was 0.0 +/- 0.0 at P3 and 0.2 +/- 0.1 at I1. Tetracycline-labeling disclosed, in the frontal cut surface of maxillofacial bones at P3, that traction affected not only the periodontal tissue and alveolar processes, but also the neighboring bones and midpalatal suture. The ratio of the labeled area in the bone, measured using an image analyzer in 6 sectors of the cut surface, was 1.2 +/- 0.3% in the left upper sector to 13.0 +/- 4.3% in the right lower sector. In the control group, it was 1.2 +/- 0.4% in the left upper and 4.8 +/- 1.1% in the right lower sector. In conclusion, vertical traction of teeth induces a vertical deformation of the maxilla. PMID- 8248919 TI - Reduction by niceritrol treatment of serum lipoprotein(a) in normolipidemic patients with coronary artery disease. AB - The effect of niceritrol at an ordinary dose (1500 mg/day) on the serum lipoprotein(a) (Lp(a)) concentration was investigated in 25 normolipidemic patients with coronary artery disease. The serum Lp(a) level was reduced by approximately 21% after 3 months of treatment (before treatment, 30.3 +/- 4.1 mg/100 ml; during treatment, 22.6 +/- 2.4 mg/100 ml; p < 0.01). By one month after the drug was discontinued, the Lp(a) returned to a pretreatment level (30.8 +/- 2.8 mg/100 ml). The levels of LDL-cholesterol and apoB were decreased significantly by the drug therapy; LDL-cholesterol increased closely to a pretreatment level after withdrawal of the drug. The percentage reduction of Lp(a) was significantly correlated with that of fibrinogen (r = 0.763, p < 0.01). Plasma concentration of fibrinogen was decreased in the patients whose reduction rate of Lp(a) was 30% or more. These results indicate that niceritrol has Lp(a) lowering effect; sufficient reduction of Lp(a) may improve dyscoagulopathy of patients with coronary artery disease. PMID- 8248920 TI - Clinical and pathological investigation of microscopic mole. AB - To clarify the pathological significance of the quasi-mole lesions of the placenta, morphological and morphometrical study of villi was undertaken. The material comprised products of conception in 70 cases of artificial abortion, 110 cases of spontaneous abortion and 11 cases of complete mole: The artificially aborted material was regarded as having normal villi. The products, after being observed with a stereo microscope, were subjected to microscopic examination of paraffin sections. A vascular villi with central liquefaction and proliferation of trophoblasts were found not only in the group of overt mole but also in spontaneously aborted and even in artificially aborted material, though less frequently. By morphometry, villi were shown to be involved in these changes even when macroscopically there seemed to be no significant abnormalities, a condition we prefer to call "microscopic mole." This was prevailing particularly in cases in which the average minor axis of villi exceeded 1 mm; it became more predominant with enlarging axis, gradually approaching the shape of complete mole. Some of the cases with enlarged villi proved to develop later invasive mole. Thus, whether or not the average minor axis exceeds 1 mm is likely to have a profound clinical significance. PMID- 8248921 TI - Enhanced lymphatic delivery of monoclonal antibody following OK432 pretreatment. AB - The effect of OK432 on the lymphatic delivery of monoclonal antibody (Mab) was investigated by injecting [125I]-Mab A7 into BALB/c mice pre-treated with OK432, and measuring the radioactivity in the regional lymph nodes. The [125I]-Mab A7, when administered subcutaneously to the foot pad, preferentially accumulated in the ipsilateral popliteal and para-aortic lymph nodes. Treatment with OK432 prior to antibody injection significantly enhanced the accumulation of Mab A7 in these regional lymph nodes. These findings suggest that pretreatment with OK432 may constitute a promising method of enhancing the lymphatic delivery of Mab, and improving the efficacy of therapy directed against lymphatic malignancies. PMID- 8248922 TI - Psychoses in three cases with myasthenia gravis and thymoma--proposal of a paraneoplastic autoimmune neuropsychiatric syndrome. AB - Three patients with neuropsychiatric symptoms (NPSs) associated with thymoma, high serum titers of antiacetylcholine receptor (AchR) antibody and generalized myasthenia gravis (MG) are reported. The NPSs were homogeneous; (1) Altered consciousness as manifested by dreamy state with paramnesia, (2) psychosensory symptoms (the sudden change of senses of smell and taste with behavior abnormalities, auditory and visual hallucinations, deja experiences, microteleopsia and derealization), (3) cognitive disturbances (recent memory loss with compulsive behaviors), (4) emotional disturbances (agitation, fear and anger), and (5) psychotic symptoms (secondary delusions and hallucinations) were characteristic. The NPSs preceded by several months to years the onset of MG, and thereafter they were closely related to worsening and relapse of MG. A typical patient showed repeatedly abnormal electroencephalograms (EEG) indicative of cerebral dysfunction. Another showed improvement of the NPSs after thymectomy and immunosuppressive therapy. The organicity of the phenomenology of psychoses with the same NPSs was suggested and it appears to comprise a unique paraneoplastic syndrome by central autoimmune mechanism. We proposed an autoimmune psychiatric syndrome and the genesis of psychosis due to the central cholinergic dysfunction in MG. PMID- 8248923 TI - Morbidity of allergic rhinitis based on the National Health Insurance records of Japan. AB - The morbidity of allergic rhinitis (AR) based on Japan National Health Insurance records was calculated from 1981 to 1990. The data shows a three-fold increase of the rate for AR patients in those 10 years. Additionally, Aichi Prefecture's morbidity of AR was analysed for May 1991. When cross-sectional methods were used for May 1991, it was found that the morbidity of AR showed little correlation with the pollen count for Japanese cedars and Japanese cypresses. However, the results suggested possible correlations between the morbidity of AR and the mean yearly levels of the pollutant components SPM and NO2. PMID- 8248924 TI - Adhesion and motility of polymorphonuclear leukocytes isolated from the blood of rats exposed to ozone: potential biomarkers of toxicity. AB - Ozone (O3) exposure of rats results in airway epithelial injury and an infiltration of polymorphonuclear leukocytes (PMNs) into the lungs, suggesting alteration of PMN functions. To identify the altered PMN functions and their possible effects on epithelia, rats were exposed to air or 0.8 ppm O3 for 2 hr. PMNs were isolated from the blood and incubated with an epithelial cell line derived from rat lung (ARL-14) or primary alveolar type II cell cultures. The PMNs from the O3-exposed rats exhibited stimulated motility and spontaneous redistribution of actin filaments and adhered in a greater number to the epithelial cells when compared with the PMNs from the air-exposed rats. Actin caps usually formed at the sites of contact between the PMNs and epithelial cells, suggesting a cytoskeletal role in the inflammatory-epithelial cell interaction. By scanning electron microscopy, PMNs from air-exposed rats had features of non-motile cells. In a striking contrast to this, PMNs from O3 exposed rats revealed surface modifications, which were quite prominent at the sites of PMN-epithelial cell contacts. Despite these morphological changes, the PMNs from O3-exposed rats did not alter the epithelial resistance, a measure of paracellular permeability. In contrast to this, PMNs stimulated by phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine not only exhibited greater adhesion to the epithelial cells, but also caused a reduction in epithelial resistance. The changes reflecting altered morphology, motility, and adhesion of PMNs from O3-exposed rats may represent important steps in the O3 induced inflammatory response that precedes barrier disruption in vivo, but they are not associated with increased epithelial permeability in an in vitro system. Besides their mechanistic relevance, the alterations of vascular PMNs may serve as important biomarkers for detecting O3 effects. PMID- 8248925 TI - In vivo ESR spin trapping evidence for hydroxyl radical-mediated toxicity of paraquat and copper in rats. AB - The electron spin resonance (ESR) spin trapping technique was employed to study the role of Cu(II) in the generation of hydroxyl radical during paraquat (PQ2+) intoxication in rats. A secondary radical trapping technique was used for the detection of hydroxyl radical generated in vivo during copper-mediated PQ2+ toxicity. According to this technique (Burkitt and Mason, Proc. Natl. Acad. Sci. USA 88, 8440-8444, 1991), the hydroxyl radical generated reacts with dimethyl sulfoxide (DMSO) in vivo to form the methyl radical, which is then spin trapped by phenyl-N-tert-butylnitrone (PBN). The relatively stable methyl radical adduct of PBN was detected in the bile of rats 2 hr after treatment with simultaneous doses of CuSO4, PQ2+, PBN, and DMSO, whereas no radical adducts were detected in the absence of administered PQ2+. Bile samples were collected into Cu(I)- and Fe(II)-stabilizing agents in order to prevent the occurrence of radical adducts generated ex vivo in bile during its collection. The analysis of radical adducts excreted via the biliary route provide strong ESR evidence for the generation of the hydroxyl radical as a result of the known futile enzymatic redox cycling of PQ2+, with copper playing an essential mediatory role. No radical adducts were detected when either CuSO4 or PQ2+ was excluded. From a different perspective, in vivo copper-dependent hydroxyl radical generation could be said to be promoted by PQ2+. This is the first report of ESR evidence for this synergetic hydroxyl radical generation by copper and PQ2+ in a whole animal. PMID- 8248926 TI - CYP4B1 activates 4-ipomeanol in rat lung. AB - Inhibition of pulmonary CYP4B1 activity by pretreatment of rats with p-xylene decreased the ability of lung microsomes to N-hydroxylate 2-aminofluorene and prevented the lung damage normally seen after dosing with ipomeanol. The toxicity of ipomeanol, as assessed by acute lethality, was decreased by a factor of eight. In contrast, induction of CYP1A1 by Aroclor or beta-naphthoflavone, or inhibition of CYP2B1 by O,O,S-trimethyl-phosphorodithioate, as assessed by measurement of lung microsomal dealkylation of ethoxyresorufin or pentoxyresorufin, did not change ipomeanol toxicity. A polyclonal antibody raised against CYP4B1 prevented the covalent binding of [14C]-ipomeanol to lung microsomal protein in vitro. Antibodies raised against the other major P450 isozymes of rat lung, CYP2B1 and CYP1A1, had no effect on this binding. Aroclor, beta-naphthoflavone, and O,O,S trimethylphosphorodithioate failed to affect binding of radiolabeled ipomeanol in vivo, but pretreatment with p-xylene resulted in a significant reduction in this binding. The CYP4B1 substrate 2-aminofluorene, when dosed to rats, caused a sixfold decrease in ipomeanol toxicity. These results indicate that in the rat, unlike the rabbit, pulmonary bioactivation of ipomeanol is predominantly dependent upon CYP4B1. PMID- 8248927 TI - The behavioral syndrome caused by 3,3'-iminodipropionitrile and related nitriles in the rat is associated with degeneration of the vestibular sensory hair cells. AB - Animals exposed to 3,3'-iminodipropionitrile (IDPN) or to several similar nitriles develop a permanent syndrome of behavioral abnormalities. The present work addressed the hypothesis that this syndrome is caused by a toxic effect of these nitriles on the peripheral vestibular system. Male Long-Evans rats were given acute doses of IDPN (0, 200, 400, 600, or 1000 mg/kg, ip) and assessed for a number of behaviors indicative of vestibular function at postdosing times ranging from 1 day to 9 weeks. The pathological effects of IDPN on the morphology of the vestibular sensory epithelia were studied by scanning electron microscopy at 1,2,4, and 21 days after exposure. The behavioral study revealed dose dependent deficits in vestibular function after IDPN. Alterations in vestibular morphology occurred at the same doses of IDPN that induced behavioral changes (400-1000 mg/kg). The pathological alterations after IDPN consisted of degeneration of the vestibular sensory hair cells, and no hair cells remained in the vestibular receptors 3 weeks after the 1000 mg/kg dose. A good correlation was also found for the time-course characteristics of the behavioral and the morphopathological effects of IDPN. The vestibular sensory epithelia displayed a regional pattern of differential sensitivity to the toxic effect of IDPN. Both intraepithelial and interepithelial differences in sensitivity were found. Crotonitrile (250 mg/kg, ip), which induces the same behavioral syndrome, was found to induce also degeneration of the vestibular hair cells. We conclude that IDPN and the similar nitriles that cause the same behavioral abnormalities are toxic to the peripheral vestibular system. PMID- 8248928 TI - A comparison of the pulmonary defenses against streptococcal infection in rats and mice following O3 exposure: differences in disease susceptibility and neutrophil recruitment. AB - Ozone (O3) exposure reduces alveolar macrophage (AM) phagocytosis in mice and increases their susceptibility to Streptococcus zooepidemicus. O3 exposure also decreases AM phagocytosis in rats but does not result in mortality to infection. To investigate the mechanism of disease protection in rats, antibacterial defenses of two strains of mice and F344 rats were compared. O3 exposure (3 hr, 0.4 or 0.8 ppm) and infection with S. zooepidemicus resulted in a dose-dependent proliferation of bacteria in the lungs of mice and high mortality. Polymorphonuclear leukocytes (PMNs) were observed in severely affected individuals 2 or more days postinfection and did not alter the fatal infection. In contrast, microbial inactivation was only impaired in O3-exposed rat lungs during the first 48 hr after infection. In these animals PMNs could be isolated from bronchoalveolar lavage fluid between 6 and 48 hr postinfection with the peak response occurring at 24 hr. Pretreatment with anti-PMN serum eliminated the neutrophil influx and impaired further the bactericidal activity in ozone-exposed rats. The results suggest that inhaled streptococci are cleared normally from the mouse lung by AMs. Following exposure to O3, AM phagocytosis is reduced and the mice develop a fatal infection. The persistence of bacteria in the lungs of O3 exposed rats triggers a transient influx of PMNs whose appearance corresponds with elimination of the bacteria. Differences in antimicrobial defenses between various experimental species and humans need to be better understood in order to predict effects of air pollutants on susceptibility to infection in man. PMID- 8248929 TI - Acetaminophen hepatotoxicity: influence of phenobarbital and beta-naphthoflavone treatment in obese and lean Zucker rats. AB - The effect of phenobarbital (PB) and beta-naphthoflavone (beta-NF) on acetaminophen (APAP)-induced hepatotoxicity was evaluated in obese and lean Zucker rats. In addition, the consequences of APAP overdose on hepatic CYP2B1/2B2 enzyme activities following PB treatment were assessed. A single oral dose of APAP 3 g/kg (total body weight) was administered to both littermates 24 hr after the last dose of PB or beta-NF. Histologic evidence of hepatocellular necrosis and serum hepatic aminotransferase enzymes 48 hr after APAP administration was utilized to evaluate hepatic damage. Hepatic microsomal total cytochrome P450 concentrations, alkoxyresorufin O-dealkylase activities, and 16 beta-testosterone hydroxylase activities were determined to evaluate the effect of APAP overdose on cytochrome P450 enzyme-substrate activities in the presence and absence of PB treatment. APAP overdose produced lower hepatotoxicity in the obese Zucker rat compared to lean controls. A similar trend was observed in animals treated with PB prior to APAP administration. In contrast, beta-NF treatment produced potentiation of APAP toxicity and/or death of both obese and lean Zucker rats. Generally, APAP overdose produced reduction of hepatic cytochrome P450 enzyme substrate activities. However, obese Zucker rats retained a higher percentage of their pre-APAP-treated enzyme activities which is consistent with the observation that obese Zucker rats are less affected by the hepatotoxic effects of APAP overdosage. PMID- 8248930 TI - The effect of 2,4-dinitrophenol on the metabolic rate of bobwhite quail. AB - Bobwhite quail were exposed to 2,4-dinitrophenol (DNP) in a respirometer designed to continuously monitor exchange of O2 and CO2, from which metabolic rates (MR) were estimated. After 14-16 days of acclimation to the system (temperature 22 degrees C, light cycle 8L:14D), hens received feed amended with 0, 625, or 1250 ppm DNP ad libitum for 8 days, followed by 2-5 days of feed containing no DNP. The 625 ppm treatment produced dark-period MR 31-41% higher than corresponding control values, and light-period MR 23-32% higher than controls. The 1250 ppm treatment produced dark-period MR 48-77% higher than control values, and light period MR 41-67% higher than controls. Over the 8 days of testing, the 625 ppm treatment hens expended 32% more energy than the control hens. Hens consuming 1250 ppm feed expended 60% more energy than control hens and lost most of their body fat. In general, the risk of being unable to meet energy demands for survival or reproduction would probably be substantially increased by the observed elevation in MR. PMID- 8248931 TI - Leukocytes in carbon monoxide-mediated brain oxidative injury. AB - This study was conducted with rats to assess the involvement of leukocytes in a model of CO-mediated brain injury. Myeloperoxidase activity, measured as an index of leukocyte sequestration, was found to be increased 10-fold in brain microvessel segments prepared from rats immediately or 90 min after exposure to CO. Fluorescence and light microscopic examinations revealed leukocytes in microvessels taken from CO-poisoned rats, but not in that from control rats. Studies were then conducted with rats that had been made leukopenic or treated with monoclonal anti-CD-18 F(ab')2 fragments to inhibit leukocyte adherence to the vasculature. Neither of these groups of animals exhibited the biochemical changes observed in the brains of sham-treated rats: conversion of xanthine dehydrogenase (XD) to sulfhydryl-irreversible xanthine oxidase (XO), and lipid peroxidation, at 90 min following CO poisoning. Treatment with a synthetic serine protease inhibitor, gabexate mesylate, also prevented these biochemical changes if administered immediately after CO poisoning, but the agent did not inhibit leukocyte sequestration. Rats depleted of XD and XO by a tungsten diet, and those treated with allopurinol to inhibit XD and XO, also exhibited at least a 10-fold increase in myeloperoxidase activity in microvessels immediately after CO poisoning, but only a 5-fold increase at 90 min. In vitro studies demonstrated that B2 integrin-dependent polymorphonuclear leukocyte adherence was impaired immediately following CO poisoning although the adherence molecules were expressed on the membrane surface. Adherence function normalized by 45 min. The results suggest that leukocytes are responsible for the development of biochemical changes in brain following CO poisoning, and the sequence of events is as follows: leukocyte sequestration in the microvasculature, B2 integrin dependent adherence, protease-mediated conversion of XD to XO, O2 radical dependent lipid peroxidation. PMID- 8248932 TI - Functional inhibition of leukocyte B2 integrins by hyperbaric oxygen in carbon monoxide-mediated brain injury in rats. AB - Exposure to hyperbaric oxygen [3 atmospheres absolute (ATA) for 45 min] inhibited carbon monoxide (CO)-mediated lipid peroxidation in the brains of rats by preventing the conversion of xanthine dehydrogenase to oxidase, a conversion process known to be due to the action of leukocytes. The effect was the same whether treatment was given 24 hr before or up to 45 min after poisoning. Hyperbaric oxygen did not inhibit the initial interaction of leukocytes with brain microvasculature, based on measurements of myeloperoxidase (MPO) in microvessel segments, but persistent adherence, which is due to B2 integrins, did not occur. Exposing rats to 3 ATA pressure (0.21 ATA O2) after CO poisoning had no significant effects. A progressive reduction in brain microvessel MPO titers occurred with exposure to O2 at 1, 2, or 3 ATA after CO poisoning, but 1 ATA O2 treatment did not significantly inhibit xanthine oxidase formation or lipid peroxidation. In vitro studies with polymorphonuclear leukocytes (PMN) from rats exposed to hyperbaric oxygen corroborated the absence of PMN B2 integrin function, but when these cells were stimulated they exhibited normal B2 integrin expression on their surface and also normal elastase release and superoxide radical production. Adherence functions of PMN that do not require B2 integrins appeared to remain intact after exposure to hyperbaric oxygen, as peritoneal neutrophilia in response to a glycogen challenge was not inhibited. B2 integrin function could be restored by incubating cells with 8 bromo cGMP, and incubation with phorbol ester stimulated the adherence function of both control and hyperbaric oxygen-exposed PMN. These results provide a clear mechanism for the inhibition of CO-mediated brain lipid peroxidation by hyperbaric oxygen and indicate that hyperoxia causes a discrete disturbance of PMN adherence function. PMID- 8248933 TI - Furan-induced cytolethality in isolated rat hepatocytes: correspondence with in vivo dosimetry. AB - Furan, a rodent hepatotoxicant and hepatocarcinogen, produced incubation time- and concentration-dependent decreases in the glutathione (GSH) content and viability of freshly isolated F-344 rat hepatocytes in vitro. Since furan itself did not significantly react with GSH, these data indicate the formation of a reactive metabolite of furan in hepatocyte suspensions. Treatment of the hepatocyte suspensions with the cytochrome P450 inhibitor 1-phenylimidazole delayed GSH depletion but did not alter furan-induced (4 to 12 mM) cytolethality. The furan concentrations required to produce measurable hepatocyte cytolethality in vitro within 6 hr (4 to 12 mM) were several orders of magnitude greater than the predicted maximal liver concentrations of furan in vivo following hepatotoxic doses. In order to study the mechanisms involved in the cytolethality of furan toward hepatocytes in vitro at concentrations relevant to hepatotoxicity in vivo, a hepatocyte suspension/culture system was developed that utilized furan concentrations and incubation times similar to hepatic dosimetry in vivo. Freshly isolated rat hepatocytes in suspension (in Williams' Medium E) were incubated with furan (2 to 100 microM) for 1-4 hr and placed in culture, and viability was determined after 24 hr by lactate dehydrogenase release. Furan produced cytolethality (5 to 70%) and modest GSH depletion in an incubation time- and concentration-dependent manner. Both GSH depletion and cytolethality induced by furan were prevented by 1-phenylimidazole and enhanced by acetone pretreatment of the rats. These data show that oxidation of furan by cytochrome P450 is required for GSH depletion and cytolethality, indicating that a reactive metabolite is involved in cell death. The results of this study underscore the importance of using in vivo toxicant concentrations and exposure times for in vitro mechanistic studies of chemically induced cytolethality. PMID- 8248934 TI - Kinetic analysis of furan biotransformation by F-344 rats in vivo and in vitro. AB - Furan is both hepatotoxic and hepatocarcinogenic in rats. The kinetics of furan biotransformation by male F-344 rats were studied in vivo and in vitro in order to understand target tissue dosimetry. A physiologically based pharmacokinetic (PBPK) model for furan in rats was developed from gas uptake studies using initial furan concentrations of 100, 500, 1050, and 3850 ppm. Tissue partition coefficients for furan were determined in vitro using vial equilibration techniques. Furan gas uptake kinetics in vivo were described by a single saturable process with a Vmax of 27.0 mumol/hr/250 g rat and a KM of 2.0 microM. Furan metabolism in vivo was inhibited by pyrazole. The furan PBPK model adequately simulated blood and liver furan concentrations following 4-hr inhalation exposures to 52, 107, and 208 ppm furan. The biotransformation of furan was studied in freshly isolated rat hepatocytes in vitro and compared to biotransformation in vivo. Furan biotransformation by isolated rat hepatocytes exhibited a KM of 0.4 microM and a Vmax of 0.018 mumol/hr/10(6) cells. Inhibition and induction studies indicated that cytochrome P450 was the catalyst of furan oxidation. Acetone pretreatment of the rats produced a five-fold increase in the rate of the hepatocyte oxidation of furan, suggesting an important role for cytochrome P450 2E1. The Vmax determined in hepatocytes in vitro extrapolated to 23.0 mumol/hr/250 g rat, assuming 128 x 10(6) hepatocytes/g liver. Incorporation of the in vitro hepatocyte kinetic parameters into the PBPK model for furan accurately simulated in vivo pharmacokinetics. These results suggest that freshly isolated hepatocytes are a valuable in vitro system for predicting chemical pharmacokinetics in vivo. PMID- 8248935 TI - Cytotoxicity and DNA-protein crosslink formation in rat nasal tissues exposed to vinyl acetate are carboxylesterase-mediated. AB - Vinyl acetate is used in the paint, adhesive, and paper board industries. Vinyl acetate is a nasal carcinogen in rats exposed by inhalation for 2 years to 200 and 600 ppm, but not 50 ppm. Previous studies from our laboratory suggest that rat liver microsome-activated vinyl acetate induces plasmid DNA-histone crosslinks, in vitro, through esterase-mediated metabolism. Since nasal tissues contain high levels of carboxylesterase, tumorigenesis may be related to in situ production of the hydrolysis products acetaldehyde and acetic acid. Vinyl acetate was cytotoxic to both respiratory and olfactory tissues in vitro at 50-200 mM, but not 25 mM, after 2 hr exposure. Pretreatment of rats with the carboxylesterase inhibitor, bis-(p-nitrophenyl) phosphate (BNPP), attenuated the cytotoxic effects and metabolism of vinyl acetate in both tissue types. Semicarbazide, an aldehyde scavenger, was unable to protect the tissues from vinyl acetate-induced cytotoxicity. When the metabolites were tested, acetic acid, but not acetaldehyde, was cytotoxic to both tissues. The induction of DNA protein crosslink (DPXL) formation by acetaldehyde and vinyl acetate in rat nasal epithelial tissues was detected using a sodium dodecyl sulfate/KCl precipitation technique. Endogenous crosslink levels ranged from 0.5 to 2.0% of total DNA and were considered background. Epithelial cells isolated from both olfactory and respiratory turbinates exhibited dose- and time-dependent increases in DPXL formation when exposed to 10-150 mM acetaldehyde for 1-2 hr at 37 degrees C. Similarly, respiratory and olfactory epithelial cells exposed to 5-75 mM vinyl acetate for 1-2 hr accumulated up to 12- and 15-fold higher crosslink levels than untreated cells, respectively. However, vinyl acetate appears to induce much higher levels of DPXLs at equimolar doses than acetaldehyde. This is thought to be related to stimulation of acetaldehyde-induced DPXL formation by the pH lowering effect of acetic acid production (via vinyl acetate hydrolysis). Pretreatment of the nasal turbinates with 1 mM BNPP reduced 25 mM vinyl acetate induced DPXL formation by over 75% in both tissues. These data support a hypothesis that carboxylesterase-mediated hydrolysis of vinyl acetate is necessary to generate the active intracellular cross-linking agent, acetaldehyde, and the cytotoxic metabolite, acetic acid. PMID- 8248936 TI - Mechanism of the negative inotropic action of cocaine in rat papillary muscle. AB - We studied the mechanism of the negative inotropic action of cocaine in isolated rat papillary muscles superfused with Tyrode's solution at 31 degrees C while attached to a force transducer to measure peak tension developed (PTD), maximum velocity of development of tension (Vmax T), and time to peak tension (TPT). The results show that in preparations driven at 1 Hz, 5 and 10 mg/liter cocaine had a significant negative effect only on PTD, while 20 mg/liter (58 microM) significantly depressed all three measured parameters. Similar results were obtained in preparations driven at 0.5 Hz, which developed a greater control active tension than those driven at 1 Hz. In preparations driven at 0.5 Hz, propranolol (1 microM) had no significant effect on the active tension developed nor did it modify the negative inotropic action of cocaine. Both nifedipine (1 microM) and caffeine (1.5 mM), on the other hand, significantly reduced PTD, Vmax T, and TPT. The depressant inotropic action of cocaine was accentuated by nifedipine, but was blocked by caffeine. These observations suggest that the primary site of action of cocaine was at the level of the sarcoplasmic reticulum rather than at the sarcolemma. PMID- 8248937 TI - Specific amino acids modulate the embryotoxicity of nickel chloride and its transfer to the rat embryo in vitro. AB - To investigate the effects of amino acids on the embryotoxicity and placental transfer of nickel chloride (NiCl2), Day 10 rat embryos were cultured in rat serum medium containing NiCl2 or 63NiCl2 (0.34 or 0.68 mM Ni), with or without L histidine (2 mM), L-aspartic acid, glycine (2 or 8 mM), or L-cysteine (2 mM). After 26 hr, conceptuses were assessed for survival, growth and development, and malformations. The 63Ni contents of embryos and yolk sacs and the extent of 63Ni binding to the proteins of the culture medium were also determined. NiCl2 alone did not affect the embryonic development at 0.34 mM and caused growth retardation and brain and caudal abnormalities at 0.68 mM. Coincubation of L-histidine with 0.34 mM Ni increased Ni concentrations in embryonic tissues compared to 0.34 mM 63Ni alone, but did not elicit NiCl2 embryotoxicity. Coincubation of L-cysteine with 0.34 mM Ni elicited growth retardation and brain abnormalities caused by NiCl2 and increased yolk sac concentrations of 63Ni compared to 0.34 mM 63Ni alone. In contrast, coincubation of L-histidine, L-cysteine, or L-aspartic acid with 0.68 mM Ni reduced the growth retardation and the incidence and/or severity of brain defects caused by NiCl2 and decreased the concentrations of 63Ni in the yolk sacs, compared to 0.68 mM 63Ni alone. L-Histidine also reduced the percentage of NiCl2-elicited caudal defects. Coincubation with glycine did not NiCl2-elicited caudal defects. Coincubation with glycine did not affect the embryotoxic profile, nor the placental transfer of NiCl2. In the presence of L histidine, L-cysteine, or L-aspartic acid, there was a shift of 63Ni binding from the high-molecular-weight proteins of the culture medium to the low-molecular weight fraction. Thus, specific extracellular amino acids can modulate the embryotoxicity and placental transfer of NiCl2 in vitro. The pattern of this modulation is dependent on the concentration of NiCl2, as well as on the amino acid. PMID- 8248938 TI - The effect of Ca2+, Cd2+ and Ni2+ on detergent-permeabilized vascular smooth muscle from the shark, Squalus acanthias. AB - We examined the effect of Ca2+, Cd2+, or Ni2+ on vascular smooth muscle intracellular proteins involved in contraction, using rings of detergent permeabilized aortae from the spiny dogfish, Squalus acanthias. Addition of Ca2+ stimulated contraction of the vascular smooth muscle, and permeabilization by treatment with Triton X-100 increased the sensitivity to Ca2+ nearly 5 log units, demonstrating that this protocol left contractile and regulatory proteins intact. Addition of 1 microM calmodulin did not increase the sensitivity of the rings to Ca2+, suggesting that this preparation is not leaky to this regulatory protein. Neither Cd2+ nor Ni2+ stimulated contraction of permeabilized rings demonstrating that the previously-described contractile action of these heavy metals is not mediated by direct stimulation of intracellular proteins, rather by interaction with sarcolemmal proteins. PMID- 8248939 TI - Effect of exposure to low level lead on growth and growth hormone release in rats. AB - This study was undertaken to examine the effect of exposure to low level lead on growth and growth hormone (GH) release. Female pups exposed to lead beginning in utero were smaller than controls on postnatal day 7 (P = 0.06). There was no corresponding effect in males. No overall differences in body weights were detected in either sex with respect to treatment effect. No differences in food or water intake were observed at any time. Pituitaries from 49-day-old lead treated pups responded to in vitro incubation with growth hormone releasing factor (GRF) with a smaller increase in GH release than those from control pups (P = 0.08). In the case of the dams, lead did not affect body weight, body length, food consumption or pituitary responsiveness; however, water consumption was significantly increased in the lactating dam (P < 0.05). Interestingly, blood lead content in 5-day-old pups (43.3 +/- 2.7 micrograms/dl) exposed to lead in utero was more than twice that of their 49-day-old litter-mates (18.9 +/- 0.7 micrograms/dl). At 49 days blood lead levels in female pups (19.94 +/- 0.8 micrograms/dl) were significantly higher than those of male pups (17.00 +/- 1.1 micrograms/dl). Maternal blood lead levels on the same day averaged 22.7 +/- 2.5 micrograms/dl. This study suggests that exposure to a low level of lead can reduce pituitary responsiveness to a hypothalamic stimulus. In addition, the data reinforce the importance of considering age and sex when evaluating the toxic effects of lead. PMID- 8248940 TI - Vanadium treatment of diabetic Sprague-Dawley rats results in tissue vanadium accumulation and pro-oxidant effects. AB - The effect of sodium metavanadate (NaVO3) consumption on trace element metabolism, components of the antioxidant defense system and lipid oxidative damage were studied in control (CON) and streptozotocin-induced diabetic (DIAB) rats. Ten days after injection, CON and DIAB rats received either 0 mM NaVO3/80 mM NaCl (0 group) or 1.2 mM NaVO3/80 mM NaCl (1.2V group) in their drinking water. DIAB groups had higher food and fluid intakes than the CON groups; vanadium (V) groups had lower food and fluid intakes than the saline groups. Vanadium therapy lowered plasma glucose concentrations of DIAB rats. The following parameters were similar among the groups: plasma Zn, Cu and Fe concentrations, plasma ceruloplasmin activity, liver Zn, Cu, Mn and Fe concentrations, kidney Mn and Fe concentrations, liver non-Se-dependent glutathione peroxidase (GSH-Px), glutathione reductase (GSH-Red) and Mn-SOD activities, liver reduced glutathione (GSH) and oxidized glutathione (GSSG) concentrations and kidney non-Se-dependent GSH-Px activity. Kidney Zn and Cu concentrations were higher in DIAB rats than in CON rats. The CON-1.2V and DIAB 1.2V groups had V accumulation in the liver and kidney. Liver CuZn-SOD and Se dependent GSH-Px and kidney CuZn-SOD and GSH-Red activities were lower in DIAB rats compared to CON rats; kidney Mn-SOD and kidney Se-dependent GSH-Px activities were higher in DIAB rats than CON rats. Vanadium treatment did not cause significant alterations in the antioxidant defense system; however, tissue vanadium concentrations were positively correlated to TBARS production. These results show that diabetes caused significant alterations in the antioxidant defense system and that V therapy was associated with a marked deterioration in health of both control and diabetic rats. PMID- 8248941 TI - Electron paramagnetic resonance studies of the effects of methoxyacetic acid, a teratologic toxin, on human erythrocyte membranes. AB - Methoxyacetic acid (MAA), a teratogenic toxin, is the major metabolite of ethylene glycol monomethyl ether (EGME, also referred to as 2-methoxyethanol, 2 ME). MAA causes a wide range of toxic effects in laboratory animals including reproductive and developmental toxicity, as well as hematotoxicity, by mechanisms that are not clear. In this study, we employed electron paramagnetic resonance (EPR) spin-labeling techniques in conjunction with spin labels specific for cytoskeletal proteins, bilayer lipids, cell-surface sialic acid, or cell-surface galactose and N-acetylgalactosamine residues of human erythrocyte membranes in order to gain insight into the mechanism of MAA toxicity. The major findings are: (1) MAA significantly increases the protein-protein interactions of skeletal proteins in a concentration-dependent manner (P < 0.001), while 2-ME has no effect (at even a 2.5-fold higher concentration). (2) Addition of MAA leads to significant increase in the rotational motion of spin-labeled terminal galactose and N-acetylgalactosamine residues (2.0 mM MAA, 38% decrease of the apparent rotational correlation time tau a, P < 0.01). (3) The rotational motion of spin labeled sialic acid, 70% of which is on the major transmembrane sialoglycoprotein (glycophorin A or PAS 1), was not affected by MAA treatment. (4) MAA has no effect on the lipid bilayer fluidity, since no change in the motion of a lipid bilayer specific spin label (5-NS) in the erythrocyte membrane was observed. These results suggest that MAA may lead to teratologic toxicity by interacting not with lipid components but with certain, perhaps specific, protein components, i.e., transport proteins, cytoskeleton proteins or neurotransmitter receptors. PMID- 8248942 TI - Selective removal of copper bound to metallothionein in the liver of LEC rats by tetrathiomolybdate. AB - LEC rats (Long-Evans with a cinnamon-like coat color) have a genetical defect in Cu metabolism. Male LEC rats aged 10 weeks were injected ip with TTM at a dose of 5 or 10 mg/kg body weight for 8 consecutive days and killed one day after the last injection. Cu that had accumulated in the liver at a concentration of 251 micrograms/g liver was decreased to 82.7 or 74.3 micrograms/g liver respectively, by the treatment. Although most of Cu was bound to MT as a soluble form before TTM treatment, the metal remaining in the liver after the treatment was present almost exclusively in the non-soluble fraction. Zinc (Zn) present, bound to MT before the treatment, was also partly removed from the liver by TTM, and the Zn remaining in the liver after the treatment was revealed to be bound to MT (Zn-MT) by high performance liquid chromatography-atomic absorption spectrophotometry. Iron (Fe) in the liver was not affected by TTM treatment. Cu in the kidneys and spleen increased by TTM treatment, while Zn and iron were not affected. Treatment of LEC rats with severe jaundice effectively cured the animals from otherwise lethal signs by only two ip injections of TTM at a dose of 10 mg/kg body weight. PMID- 8248943 TI - Airway epithelial labeling index as an indicator of ozone induced lung injury. AB - Rats were implanted subcutaneously with a bromodeoxyuridine-filled minipump and then were exposed to ozone delivered at a low dose rate (0.4 ppm during 12 h per night) or at a high dose rate (0.8 ppm during 6 h per night). Three and 7 days after pump implantation the cumulative labeling indices were measured in the alveolar zone and in the airways. Greater alveolar labeling indices were observed 7 days after implantation of the minipumps than after 3 days in all groups, but no ozone-related changes were found in the alveoli of rats in either experimental group at either time. After 3 or 7 days, the labeling index in the large intrapulmonary airways and in the terminal bronchioli of the rats exposed to the higher dose rate (0.8 ppm) was increased. In rats exposed to the lower dose rate (0.4 ppm) the labeling index was significantly elevated in the terminal bronchioli after 3 days and in both the terminal bronchioli and large intrapulmonary airways after 7 days. In the terminal bronchioli the extent of cell proliferation appeared to be defined by dose rate rather than by cumulative exposure. It is concluded that measurement of the airway labeling index is a sensitive indicator of the response of the rat lung to acute exposure to ozone. PMID- 8248944 TI - Subchronic toxicity of 2,3,7,8-tetrabromodibenzo-p-dioxin in rats. AB - 2,3,7,8-Tetrabromodibenzo-p-dioxin (2,3,7,8-TBDD) was administered daily to male and female rats for 91 days by gavage. Ten male and 10 female rats per group received 0.01, 0.1, 1, 3, or 10 micrograms 2,3,7,8-TBDD/kg body weight per dose per day, solubilised in arachis oil. At 1 microgram/kg per day and above, body weight gain was dose-dependently reduced by treatment. Animals in the 3 and 10 micrograms/kg dose groups showed symptoms of wasting syndrome. Fifty percent of the animals in the 3 micrograms/kg dose-group died and all animals of the highest dose (10 micrograms/kg) died or had to be killed in extremis. Hematological investigations indicated changes--mainly in the 1 and 3 micrograms/kg dose-groups -in hemoglobin content, packed cell volume and number of thrombocytes. The prothrombin-time was markedly prolonged after 3 micrograms/kg in week 13. Clinical chemistry performed at the end of treatment revealed an increase in plasma alkaline phosphatase (APh), aspartate aminotransferase, ASAT and alanine aminotransferase, ALAT (females only) in the highest surviving dose-group (3 micrograms/kg). Marginal changes of APh and ASAT were seen in rats in the 1 microgram/kg dose-group. In the same animals, total bilirubin was elevated. Triglycerides were reduced mainly at 1 and 3 micrograms/kg. Serum thyroxin was reduced, beginning with a marginal change at 0.1 micrograms/kg, triiodothyronine was elevated, starting with a dose of 1 microgram/kg. Thymus weights were reduced in rats of the 1, 3 and 10 micrograms/kg dose-groups. Histopathological analysis showed atrophy of the lymphatic tissue in thymus and spleen. Investigations of the liver indicated peliosis hepatis after treatment with 3 or 10 micrograms/kg. Activities of microsomal enzymes (ethoxyresorufin O-deethylase, ethoxycoumarin O deethylase, aryl hydrocarbon hydroxylase, UDP-glucuronyltransferase) investigated in liver, lung and kidney were dose-dependently elevated after 13 weeks of treatment. At a dose of 3.0 micrograms/kg, activities were below those of the dose 1.0 microgram/kg, probably due to liver toxicity. The induction ratio of kidney was generally higher than in liver and lung. No signs of treatment-related toxicity were observed in the 0.01 and 0.1 micrograms/kg groups after the subchronic administration of 2,3,7,8-TBDD by gavage. PMID- 8248945 TI - The interaction of 2,4-dichlorophenoxyacetic acid, ribosomes and polyamines in Azospirillum brasilense. AB - 2,4-Dichlorophenoxyacetic acid (2,4-D) is an herbicide used extensively in agriculture. We had previously determined that 1 mM 2,4-D could inhibit cell growth, DNA and protein synthesis of Azospirillum brasilense. The present work was designed to determine if these alterations are a consequence of 2,4-D action on polyamine biosynthesis and if the protein synthesis inhibition is a result of ribosomal impairment. In this paper we demonstrate that 2,4-D alters the metabolism of polyamines and, thus, affects protein synthesis at the ribosomal level. PMID- 8248946 TI - Effect of chronic cadmium exposure on glutathione S-transferase and glutathione peroxidase activities in rhesus monkey: the role of selenium. AB - The effect of cadmium (Cd) on the activity of glutathione S-transferase (GST) and glutathione peroxidase (GSH-Px) which play an important role in the detoxification of xenobiotics, was studied in the liver, kidney, heart and lung of Rhesus monkeys. Furthermore, the role of selenium (Se) in the modulation of Cd toxicity with respect to GST and GSH-Px was also evaluated. Cadmium exposure (5 mg Cd/kg body wt./day as CdCl2 for 10 weeks) to monkeys resulted in decreased GSH Px activity in all four organs present in the order liver > kidney > heart > lung. Cadmium administration also resulted in a significant decrease in total GST activity present in the order liver > heart > kidney > lung, whereas a significant increase in the pi class GST activity was observed greatest in the heart followed by lung, kidney and liver. Oral administration of Se (0.5 mg Se/kg body wt./day as Na2SeO3 for 10 weeks) caused a significant increase in GSH-Px activity in the order liver > heart > kidney > lung. Selenium administration caused an increase in total GST activity in liver and lung but a decrease in kidney and heart. Simultaneous administration of Cd and Se resulted in an increase in total GST activity (except in lung) including the pi class activity as well as GSH-Px activity in all four tissues under study. Thus, the mechanism by which selenium decreases Cd toxicity in Rhesus monkeys, seems to rely on the protection of the enzyme systems GST and GSH-Px in the four organs, possibly by forming non-toxic cadmium selenide. PMID- 8248947 TI - Cadmium stimulation of plasminogen activator inhibitor-1 release from human vascular endothelial cells in culture. AB - We investigated the effect of cadmium (0.5, 1.0 or 2.0 microM) on the release of plasminogen activator inhibitor-1 antigen (PAI-1:Ag) from cultured human vascular endothelial cells. It was found that cadmium at 1.0 and 2.0 microM significantly increased the PAI-1:Ag release from the cells after a 24-h incubation. However, the tissue plasminogen activator antigen (t-PA:Ag) release was not changed by the metal. Although nickel as well as cadmium increased the PAI-1:Ag release, the other heavy metals including cobalt, zinc and copper did not exhibit such a stimulatory effect. The incorporation of [3H]leucine into the acid-insoluble fraction of the cell layer was unchanged by cadmium, suggesting that the metal may stimulate the synthesis of PAI-1 without association with generalized increase in protein synthesis. Cadmium at 1.0 and 2.0 microM significantly decreased the t-PA activity in the medium. The present study showed that cadmium increases the release of PAI-I:Ag from cultured endothelial cells without non specific stimulation of protein synthesis. The decrease in the t-PA activity suggests an implication of cadmium in vascular lesion which is mediated by anti fibrinolytic activity. PMID- 8248948 TI - Depletion of total non-protein sulphydryl groups in mouse tissues after administration of d-amphetamine. AB - A study of total non-protein sulphydryl groups (TNPSH) depletion was performed in male Charles River CD1 mice (25-30 g) isolated or housed in pairs. The mice were injected intraperitoneally with d-amphetamine (5, 20 and 80 mg/kg), kept at 20 +/ 1 degrees C, sacrificed at different times (between 0 and 120 min) and TNPSH in brain, lungs, heart, liver, kidney and spleen was quantified. d-Amphetamine significantly reduced the TNPSH levels in the liver and kidney, the reduction being more pronounced in the latter organ. In the other organs assayed no depletion was observed. Housing in pairs increased depletion of TNPSH both in the liver and in the kidney. The dose of 20 mg/kg caused a higher TNPSH depletion in paired mice than the dose of 80 mg/kg did in isolated mice. Rectal temperatures (RT) were also determined in the different times for the same doses of d amphetamine. For the two lowest doses temperature values were similar in isolated and paired mice and were inversely proportional to TNPSH depletion in kidney. The administration of 80 mg/kg induced the highest TNPSH depletion in kidney and liver, but the hyperthermic effect was lower than that elicited by 20 mg/kg. No proportion was observed between RT and TNPSH depletion in liver. These results suggest that d-amphetamine changes TNPSH homeostasis in mouse liver by a temperature independent mechanism. In kidney, the hyperthermic effect may have some influence. Housing mice in pairs increases TNPSH depletion, but has no influence in the hyperthermic effect of d-amphetamine, with this experimental protocol. PMID- 8248949 TI - Cytochrome P450IA1 activity in human myometrium and uterine leiomyoma: influence of smoking. AB - A xenobiotic-metabolizing enzyme, the P450IA1 (aryl hydrocarbon hydroxylase; AHH) was measured fluorometrically in myometrium and in uterine leiomyoma tissues. The material included both myometrium and leiomyoma tissue samples from 62 leiomyoma patients and 22 tissue samples of normal myometrium obtained from leiomyoma-free patients undergoing hysterectomy. Among 62 leiomyoma patients there were 14 cigarette-smokers and 48 non-smokers, and among 22 control patients 6 and 16, respectively. The P450IA1 activity of the leiomyomas was significantly (P < 0.0005) higher than that of the surrounding myometrium in both smoker and non smoker groups. In smokers, myometrium an obvious induction of P450IA1 was seen. Smoking increased the P450IA1 activity also in the leiomyoma, but the increase was not statistically significant. Our study supports the idea that myometrial tissue can respond to xenobiotics from the environment. PMID- 8248950 TI - Trifluoromethylanilines--their effect on DNA synthesis and proliferative activity in parenchymal organs of rats. AB - Reactive isomeric 3- and 4-trifluoromethylanilines (3-,4-TFMA), and control aniline itself, induced the following effects on biosynthesis of DNA in the liver, kidney, thymus and spleen of rats: (a) The administration of 4-TFMA initially suppressed the utilization of labeled thymidine for splenic DNA synthesis during the early prereplicative stage. However, with progressing time the incorporation of the labeled marker began to increase and in 30 h its level exceeded the controls by more than 200%. As expected, aniline administration resulted in mild depression of incorporation during the whole period studied. (b) 4-TFMA caused a significant increase of incorporation of labeled thymidine into DNA thymine also in the thymus. After administration of aniline the utilization of labeled thymidine for the synthesis of DNA thymine in thymus was suppressed during the first 16 h. (c) The dose-response curve showed a linear increase of incorporation in the spleen within the dose range between 0.125 and 0.500 mmol/kg of 4-TFMA. (d) It appears that enhanced incorporation of labeled thymidine into splenic and thymic DNA is a phenomenon specific for compounds bearing the CF3 group on the 4-position of the phenyl ring, such as 4-TFMA and 4-TFMPD. On the contrary, the analogous 3-CF3 substituted derivatives had no effect. Increased incorporation of labeled thymidine into spleen and thymus DNA apparently represents an increased DNA synthesis and cellular proliferation in lymphatic organs. The proliferative response was possibly evoked by the preceding hemolysis or by other toxic effects caused by the drug. PMID- 8248951 TI - Mercury-metallothionein and the renal accumulation and handling of mercury. AB - In the present study, we evaluated the renal and hepatic accumulation of mercury, the intrarenal distribution of mercury and the urinary and fecal excretion of mercury in rats injected intravenously with a non-toxic 0.1 mumol/kg-dose of mercury in the form of mercuric chloride (HgCl2) or a complex of mercury metallothionein (Hg-MT). Between 6 and 72 h after injection, the concentration of mercury in the kidneys of the rats injected with Hg-MT was significantly greater than that in the rats injected with HgCl2. The greatest difference in the renal concentration of mercury between the two groups of rats was detected 6 h after injection. In the kidneys of both experimental groups of rats, the cortex and the outer stripe of the outer medulla contained the highest concentrations of mercury, with the greatest concentrations found in the renal cortex and outer stripe of the outer medulla of the rats injected with Hg-MT. No differences were found between the two experimental groups with respect to the concentration of mercury in the renal inner stripe of the outer medulla and inner medulla throughout 72 h of study. The content of mercury in the blood and liver decreased over time in both groups of rats, but was always significantly greater in the blood and liver of rats injected with HgCl2. The rats injected with Hg-MT excreted more than eight times the amount of mercury in the urine than the corresponding rats injected with HgCl2 during 72 h. These data indicate that there may be decreased tubular reabsorption of filtered Hg-MT and/or tubular secretion of mercury in the rats injected with Hg-MT. In contrast, the rats injected with HgCl2 excreted significantly more mercury in the feces during the same period of time than the corresponding rats injected with Hg-MT. In conclusion, our data clearly indicate that the renal and hepatic uptake and accumulation of mercury, and the urinary and fecal excretion of mercury, are altered significantly when inorganic mercury is administered intravenously as a complex with metallothionein. PMID- 8248952 TI - The effect of hydroxyl-containing dithiocarbamates on cisplatin-induced long-term renal dysfunction in Wistar rats. AB - The possibility of a slow, longer term deterioration in renal function following the administration of cisplatin has been little studied in animal models. To obtain data on this we have examined renal function and histopathology at 30 days post i.p. cisplatin (5 mg/kg) treatment in female Wistar rats with and without the administration of hydroxyl-containing dithiocarbamates as a protective measure. In contrast to the studies terminated at shorter times, the degree of protection furnished by the use of dithiocarbamates at longer times post treatment is less impressive. Results suggest that a continuing deterioration in renal function occurs at times greater than 1 week post-treatment when such dithiocarbamates are administered at 1 and 3 h post cisplatin. This was evidenced in both several measures of renal function and in the histopathology of the S3 segment of the proximal tubule. PMID- 8248953 TI - Subacute toxicity of piperonyl butoxide in ICR mice. AB - Piperonyl butoxide, alpha-[2-(2-butoxyethoxy)ethoxy]-4,5-methylenedioxy-2 propyltol uene, is a pesticide synergist. ICR mice of both sexes were maintained on diet containing 0, 0.1, 0.3 or 0.9% of piperonyl butoxide for 20 days. At the end of the experimental period, they were necropsied. Selected organs were weighed and serum chemistries were analyzed. In male and female mice of the 0.9% group, body weight, kidney and spleen weight were depressed in comparison to those of control group. Liver weight of the 0.3 and 0.9% group of both sexes were significantly higher than those of control group. Mice of the 0.9% group of both sexes had increased serum levels of cholesterol, total protein, gamma-glutamyl transpeptidase. Histological examination of livers from mice of the 0.9% group by light microscopy showed enlarged hepatocytes, anisonucleosis and single cell necrosis. The results indicated that subacute toxicity of piperonyl butoxide in ICR mice was directed primarily at liver. PMID- 8248954 TI - One-year reduction and longitudinal analysis of carotid intima-media thickness associated with colestipol/niacin therapy. AB - BACKGROUND AND PURPOSE: The Cholesterol Lowering Atherosclerosis Study has reported significant reduction of coronary artery disease and of carotid arterial intima-media thickness (IMT) at 2 and 4 years with colestipol/niacin therapy. We now report on treatment effects on carotid IMT at 6 months and 1 year. METHODS: One hundred eighty-eight nonsmoking men, aged 40 to 59 years, with prior coronary artery bypass graft surgery were randomized to colestipol/niacin plus diet therapy or placebo plus diet therapy. Computerized image processing of carotid ultrasound films was used to measure IMT in the right common carotid artery. Treatment group comparisons were made at 6 months and 1 year (46 and 33 subjects, respectively, with baseline and 6-month or 1-year ultrasound measures). The time course of the treatment effect on carotid IMT was estimated using the complete sample of 78 subjects with baseline and on-trial data. RESULTS: No significant treatment group differences on carotid IMT were found at 6 months. At 1 year, the treated group showed significant reduction of carotid IMT (P = .01 between groups). The placebo group showed continuing progression of IMT during the 4-year study period (estimated progression rate, 0.018 mm/y). The treated group showed reduction of IMT during the first 3 years and a plateau during the remainder of the study. CONCLUSIONS: Reduction of carotid IMT was found with aggressive lipid lowering therapy. Ultrasound measures of IMT offer a noninvasive and precise measure of early carotid atherosclerosis that will decrease sample size requirements, potentially decrease dropout rates, and widen the study population of antiatherosclerotic clinical trials. PMID- 8248955 TI - Cerebral glucose metabolism as a predictor of rehabilitation after ischemic stroke. AB - BACKGROUND AND PURPOSE: Permanent neurological deficits after ischemic stroke are primarily determined by the location and size of an infarct, but social recovery and rehabilitation also depend on the functional status of brain tissue outside the infarct. Since neuronal loss and functional deactivation in peri-infarct tissue are reflected as changes in flow and metabolism, measurement of glucose consumption may yield an additional measure of rehabilitative capacity. METHODS: Seventy-six nondiabetic patients (48 men, 28 women; mean age, 56.7 +/- 14.37 years) with a first unilateral supratentorial ischemic infarct were consecutively enrolled. At stable neurological and clinical condition 9 +/- 7.2 days after the attack, cerebral metabolic rate of glucose (CMRglu) in noninfarcted brain regions was measured by positron emission tomography of fluorodeoxyglucose. Outcome was assessed 21 to 77 (mean, 50.5 +/- 11.7) months after the stroke with a rehabilitation index for daily life activities. RESULTS: At time of assessment of outcome 16 patients had died (score, 0), 22 were completely recovered (score, 200), and 38 had partially improved (rehabilitation score, 5 to 195). Younger age, absence of arterial hypertension and cardiac disease, but also higher global, ipsilateral, and contralateral CMRglu were significantly related to a better final outcome (P = .001), whereas sex and neurological deficits in the subacute stage after stroke were not related to final outcome. To evaluate the significance of CMRglu further after adjustment for clinical prognostic variables, a multiple regression analysis of the effect of age and CMRglu on rehabilitation score in homogeneous subgroups of partially recovered patients was performed. It revealed a significant positive correlation of CMRglu (P = .016) with recovery in hypertensive subjects, while age was the dominant prognostic factor (P = .07) in patients with normal blood pressure. CONCLUSIONS: These results demonstrate that outcome after stroke is significantly influenced by several factors incapacitating brain function in addition to the ischemic attack. In addition to age and cardiac disease, hypertension is an important factor leading to widespread arteriopathy with neuronal loss and tissue damage. The significant correlation of CMRglu outside the infarct with functional recovery in hypertensive subjects probably reflects the extent of hypertensive tissue damage and subsequently reduced capacity to compensate for the focal ischemic insult. PMID- 8248956 TI - Upper limb somatosensory evoked potentials as a predictor of rehabilitation progress in dominant hemisphere stroke patients. AB - BACKGROUND AND PURPOSE: The aim of this study was to determine the predictive yield of upper limb short latency somatosensory evoked potential (USEP) in patients with first stroke in the dominant hemisphere. METHODS: Nineteen patients (average age, 58 years) were evaluated twice: on arrival at the rehabilitation center, approximately 3 weeks after the stroke, and again approximately 10 weeks later. The clinical assessment included a quantitative evaluation of motor ability, independence in activities of daily living, and communication ability. USEP was recorded during the week of the initial clinical evaluation. Special attention was paid to the relations between USEP parameters and the dynamics of the clinical condition. RESULTS: The seven patients in whom no cortical potential could be detected showed the worst outcomes; however, the existence of cortical potentials in the remaining 12 patients did not provide a precise prediction of their "rehabilitative capacity" (ie, the extent of their progress). A correlation was established between the amplitude of the potentials recorded over both hemispheres and changes in communication ability. Additional findings included an association between shortened central conduction time over the damaged hemisphere during the first month after stroke and improvement in motor ability. CONCLUSIONS: USEP can serve as an adjuvant tool for predicting the recovery progress of stroke patients. PMID- 8248957 TI - Predicting length of stay, functional outcome, and aftercare in the rehabilitation of stroke patients. The dominant role of higher-order cognition. AB - BACKGROUND AND PURPOSE: Research in recent years has revealed factors that are important predictors of physical and functional rehabilitation: demographic variables, visual and perceptual impairments, and psychological and cognitive factors. However, there is a remaining uncertainty about prediction of outcome and a need to clinically apply research findings. This study was designed to identify the relative importance of medical, functional, demographic, and cognitive factors in predicting length of stay in rehabilitation, functional outcome, and recommendations for postdischarge continuation of services. METHODS: The influence of these factors was determined by comparing diagnostic, medical, demographic, functional, and neuropsychological information that was retrospectively obtained by reviewing the records of 86 patients admitted for comprehensive rehabilitation due to stroke (n = 36) or orthopedic injury (n = 50). Multiple linear regression with statistical adjustment to control for overprediction of variance was used to predict outcomes. RESULTS: The study revealed the primary importance of higher-order cognitive impairments (comprehension, judgment, short-term verbal memory, and abstract thinking) in extending length of stay and increasing referrals for outpatient therapies and home services after discharge for the cerebrovascular accident patients in comparison with orthopedic cohorts. CONCLUSIONS: The need is discussed for early, comprehensive assessment of deficits in cognition that affect a stroke survivor's ability to participate in a rehabilitation program and remediation that facilitates functional improvement by building on residuals of impaired abilities or teaching compensatory behaviors. PMID- 8248958 TI - Interobserver reliability of a clinical classification of acute cerebral infarction. AB - BACKGROUND AND PURPOSE: The Oxfordshire Community Stroke Project (OCSP) clinical classification of subtypes of cerebral infarction (total and partial anterior circulation infarction, lacunar infarction, and posterior circulation infarction) can be used to predict early mortality, functional outcome, and whether the infarct was likely due to large- or small-vessel occlusion. The OCSP classification was originally developed and tested by neurologists as part of a community-based study of first-ever stroke, in which some cases were seen after the acute phase. We examined the interobserver reliability of the classification when used in everyday clinical practice in patients seen during the acute phase of stroke shortly after admission to the hospital. METHODS: Two clinicians independently assessed consecutive patients admitted to the hospital with an acute stroke and recorded both the neurological features and their opinion of the subtype of infarct. RESULTS: Eighty-five patients were assessed. Interobserver agreement for the classification was moderate to good (kappa = 0.54; 95% confidence interval, 0.39 to 0.68). Differences in the assessment of the commonly elicited neurological signs explained many of the disagreements: interobserver agreement was good for some signs (hemiparesis [kappa = 0.77], dysphasia [kappa = 0.70]), moderate for some (hemianopia [kappa = 0.39]), and poor for others (sensory loss [kappa = 0.15]). CONCLUSIONS: The classification was simple and practicable (and could be widely used in routine clinical practice, randomized controlled trials, and audit), and interobserver reliability was satisfactory. PMID- 8248959 TI - Factors associated with early presentation of acute stroke. AB - BACKGROUND AND PURPOSE: Patients with stroke symptoms commonly delay many hours before seeking medical attention. We sought to explore the factors associated with early presentation of stroke patients to physicians. METHODS: We prospectively studied 100 consecutive acute stroke patients presenting to three large, urban medical centers. Using a standardized, structured interview and chart review, we assessed patient education about stroke, risk factors, clinical features of the stroke, source of stroke recognition, and timing of presentation. We did not study the distance from the site of stroke onset to the site of physician contact. RESULTS: Stroke onset time was known in 96 of the patients. Mean patient age was 71.3 years, 79% had at least one stroke risk factor, 26% had prior transient ischemic attack, 19% had prior stroke, 74% had some high school education, and 86% had regular physicians. Only 8% had been previously educated about stroke symptoms. Eighty one percent of strokes were ischemic. The mean time to physician contact was 13.4 +/- 2.3 hours (median, 4.0 hours) and to neurologist contact was 21.2 +/- 2.9 hours. A skewed distribution of presentation times accounts for the mean-median differences. A small number of patients presenting very late could have an effect on the correlations between presentation time and the variables studied. Early presentation time was associated with increased age, the sudden onset of a stable deficit, and recognition that the symptoms signified stroke. Only the sudden onset of a stable deficit correlated independently with early presentation time (P = .0048). There was no correlation between presentation time and prior transient ischemic attack or stroke, headache, vomiting, loss of consciousness or seizures at onset, or stroke subtype, but a type II error could not be excluded. CONCLUSIONS: Despite their education level, regular health care, and risk factors, especially prior stroke and transient ischemic attack, these patients were not knowledgeable about stroke and delayed many hours before contacting physicians. The course of symptoms and recognition that they signified stroke were associated with earlier presentation. Patient education focused on groups at risk may hasten the presentation and treatment of acute stroke. PMID- 8248960 TI - Relation of laboratory and clinical variables to the grade of carotid atherosclerosis. AB - BACKGROUND AND PURPOSE: To clarify diagnostic entities in ischemic stroke we analyzed the relation between the severity of carotid atherosclerosis, coagulation parameters, lipoproteins, neurological status, and risk factors in 232 patients. METHODS: Duplex ultrasonography, computed tomography scan, and laboratory investigations were performed between the third and tenth days after stroke. Based on carotid ultrasound scores, we categorized the patients into four groups (A, B, C, and D) according to severity of atherosclerosis. Corresponding laboratory variables and clinical data were statistically analyzed. RESULTS: Ultrasound scores were significantly (P < .05) higher in the male (n = 126) versus female (n = 106) patients. The hematocrit was significantly higher and thrombin time was significantly shorter in the male group compared with the female group. Severe atherosclerosis (group C) and occlusion (group D) of the internal carotid artery was associated with smoking (C = 56%; D = 78%), hypertension (C = 43%; D = 35%), claudication (C = 13%; D = 5%), and antecedent myocardial infarction (C = 9%; D = 13%). There was no statistical correlation between ultrasound scores and the patients' neurological condition. Cholesterol and plasma fibrinogen levels were significantly higher and high-density lipoprotein cholesterol was significantly lower in groups with severe atherosclerosis compared with patients with slight intimal damage. The presence of multiple plaques or thrombosis of the internal carotid artery was concordant with the prevalence of single cerebral infarcts. CONCLUSIONS: Severity of carotid atherosclerosis corresponded well with the following factors: age, smoking, and low concentration of high-density lipoprotein cholesterol. Elevation of plasma fibrinogen combined with a loss of high-density lipoprotein cholesterol is strongly associated with severe atherosclerosis and results in brain infarction. PMID- 8248961 TI - Evaluation times for patients with in-hospital strokes. AB - BACKGROUND: Each year at least 35,000 people suffer a stroke while hospitalized, but little is known about the clinical characteristics of such patients or how rapidly they are identified and evaluated. With a recent emphasis on the very early treatment of stroke, in-hospital stroke patients may be candidates for some early interventions. METHODS: This was a retrospective study using the stroke registries at two academic medical centers. Data were collected about the clinical characteristics of patients with an in-hospital stroke and who recognized the stroke. Detailed time data were analyzed to determine the time of stroke recognition, medical evaluation, and neurological evaluation. These specific time epochs were analyzed to determine which were responsible for any delays in stroke identification and assessment. Data were analyzed using nonparametric methods, including the Wilcoxon rank sum and Kruskal-Wallis procedure. RESULTS: Sixty-three patients were identified with in-hospital strokes and adequate time data. In-hospital stroke patients were recognized most frequently by nurses (63%) and by the patient (16%). Patients on a cardiology service and general surgery service accounted for 48% of all in-hospital strokes. The mean and median times from stroke recognition to a neurology evaluation were 14.5 and 2.5 hours, respectively. Total delays differed significantly with service and locale (P = .004). Patients on the Duke neurology service were evaluated significantly sooner (median delay, 0.5 hour) than patients on the Duke medical (median delay, 5.8 hours) or Duke surgical (median delay, 20.5 hours; P < .01 by Wilcoxon rank sum) services. Patients on the Yale surgical service were evaluated significantly sooner than patients on the Duke medical (P = .0006) or surgical (P = .0001) services. The time between physician notification and calling for a neurology evaluation accounted for > 60% of the total time delay for patients not on a neurology service. CONCLUSIONS: A substantial number of in hospital stroke patients experience a long delay between symptom recognition and a neurological evaluation. While medical personnel are usually notified very soon after an in-hospital stroke is recognized, such patients often do not receive a rapid neurological evaluation. Additional education of hospital staff may reduce these time delays. PMID- 8248962 TI - A model for management of patients with stroke during the acute phase. Outcome and economic implications. AB - BACKGROUND AND PURPOSE: The purpose of the study was to develop a clinical pathway for patients with nonhemorrhagic stroke during the acute hospital phase to improve the quality of care and reduce costs. METHODS: The pathway included standard admission orders and a swallow screen on day 1 of hospitalization. Physical therapy, occupational therapy, speech therapy, and social worker assessments were done on day 2. A physiatry consult was performed on day 3 if indicated, and by day 4 a discharge target date and disposition were addressed. RESULTS: Outcomes for 121 patients during the first year of pathway implementation are reported. The average length of stay on the acute service decreased from 10.9 days to 7.3 days (P < .05), reducing the charges per patient by 14.6%. Complications in the form of urinary tract infections and aspiration pneumonia rates decreased by 63.2% (P < .05) and 38.7%, respectively. CONCLUSIONS: We conclude that the implementation of a clinical pathway for patients with acute, nonhemorrhagic stroke resulted in a significant reduction in length of stay, charges, and complications while improving the quality of care. PMID- 8248963 TI - Regular light-to-moderate intake of alcohol and the risk of ischemic stroke. Is there a beneficial effect? AB - BACKGROUND AND PURPOSE: To evaluate the association between different patterns of alcohol consumption and the risk of ischemic stroke in young or middle-aged men. METHODS: One hundred fifty-six patients and 153 control subjects were included in this case-control study. The pattern and the estimated average weekly intake of alcohol were assessed using a structured questionnaire. The pattern of drinking was defined as regular (daily or almost daily) or irregular (up to three times per week), and the weekly amount of consumption was defined as nondrinking, light to-moderate drinking (up to 150 g/wk), moderate drinking (> 150 to 300 g/wk), and heavy drinking (> 300 g/wk). Multiple stepwise logistic regression models were used, and adjustments were carried out for potential confounders. RESULTS: Heavy alcohol intake associated with an increased risk of stroke (odds ratio, 4.45; 95% confidence interval, 1.09 to 18.1), whereas the risk tended to be reduced in light-to-moderate drinkers (odds ratio, 0.54; 95% confidence interval, 0.28 to 1.05). Accounting for the pattern of alcohol intake in addition to the average weekly amount in grams, regular light-to-moderate drinking showed a significant inverse association with stroke (odds ratio, 0.12; 95% confidence interval, 0.02 to 0.65), and an irregular pattern of consumption attenuated this association. Based on the same multivariate analyses, other significant independent risk factors for stroke were arterial hypertension, coronary heart disease, and history of snoring, whereas the contributions of age, diabetes mellitus, smoking, and body mass index proved to be nonsignificant. CONCLUSIONS: Light-to-moderate alcohol intake appears to have an inverse association with the risk of ischemic stroke. The beneficial effect appears to be most prominent if the consumption of alcohol is regular and evenly distributed throughout the week, whereas a sporadic or an occasional pattern of drinking seems to weaken the association. This study also supports the role of heavy drinking as an independent risk factor for ischemic stroke. PMID- 8248964 TI - Stroke subtypes among Hispanics living in Guayaquil, Ecuador. Results from the Luis Vernaza Hospital Stroke Registry. AB - BACKGROUND AND PURPOSE: Racial differences in stroke subtypes have been documented. Asians have a higher rate of cerebral hemorrhage than whites; however, there is little information about stroke subtypes among Hispanics. The purpose of this study was to determine the patterns of stroke subtypes in a population of Hispanics. METHODS: Five hundred consecutive patients with a first stroke were included. Patients were collected from hospital wards, the emergency department, and the outpatient clinic to ensure inclusion of patients with a wide range of stroke severity. Computed tomography was available in all cases. Patients with pure subarachnoid hemorrhage were excluded. RESULTS: There were 313 (62.6%) patients with an infarct and 187 (37.4%) with a hemorrhage. Hypertensive arteriolopathy was the most common cause of both infarcts and hemorrhages. The carotid territory was involved in 70.6% of the 313 patients with infarcts, the vertebrobasilar territory in 17.9%, multiple territories in 6.7%, and a watershed area in 4.8%. Hemorrhages were most often lobar (36.4%), followed by putaminal (30.5%), brain stem (9.1%), cerebellar (8%), thalamic (8%), ventricular (5.3%), and caudate (2.7%). CONCLUSIONS: This hospital-based stroke registry suggests that stroke in Hispanics has a pattern different from that in whites but similar to that in Asians. Cerebral hemorrhages occur three times more frequently in Hispanics than in whites. PMID- 8248965 TI - Early carotid atherosclerosis in healthy middle-aged women. A follow-up study. AB - BACKGROUND AND PURPOSE: Few longitudinal data about early atherosclerotic lesions of the carotid arteries are available in general populations. The main purpose of this study was to investigate risk factors for development and regression of intimal-medial thickening and atheromatous plaques. METHODS: Initial and 2-year examinations of the carotid arteries with high-resolution B-mode ultrasonography were performed in 308 apparently healthy women aged 45 to 55 years. The development of new atheromatous plaques and new intimal-medial thickening and the disappearance of preexisting plaques and intimal-medial thickening defined the four outcomes of interest. RESULTS: The development of plaques occurred more frequently in women with intimal-medial thickening than in women with normal carotid arteries at baseline (14.4% versus 7.2%, P < .053). A regression was seen in 21.7% of the women with preexisting plaques. Development of intimal-medial thickening occurred in 47.5% of the women with normal carotid arteries whereas 20.2% of the women with preexisting intimal-medial thickening showed a regression of their lesions. Multiple logistic regression showed that smoking (regression coefficient +/- SE: 1.281 +/- 0.450; P < .005), baseline levels of systolic blood pressure (regression coefficient +/- SE: 0.031 +/- 0.015; P < .04) and apolipoprotein B (regression coefficient +/- SE: 0.016 +/- 0.007; P < .03) were independently associated with the development of plaques, whereas the presence of an intimal-medial thickening did not reach the significance level (regression coefficient +/- SE: 0.639 +/- 0.436; P < .15). Independent predictors of the development of intimal-medial thickening were age (regression coefficient +/- SE: 0.124 +/- 0.048; P < .04) and, with a borderline significance level, (log)triglycerides (regression coefficient +/- SE: 0.854 +/- 0.451; P < .06). Low levels of low-density lipoprotein cholesterol (regression coefficient +/- SE: 0.027 +/- 0.009; P < .004) were associated with its regression. CONCLUSIONS: This longitudinal study emphasizes the interest of B-mode ultrasonography in the monitoring of early carotid lesions. It gives further support to the hypothesis that intimal-medial thickening may be an early indicator of the atherosclerotic process. PMID- 8248966 TI - The J-curve phenomenon in stroke recurrence. AB - BACKGROUND AND PURPOSE: The relation of poststroke blood pressure to stroke recurrence remains undetermined, and the optimal control of blood pressure has not been established. We performed the present study to resolve these issues. METHODS: We analyzed 368 stroke patients with a history of hypertension (mean age, 62 years) who were admitted within 3 months after stroke onset and observed for 6 months or more. We determined stroke recurrence rate in relation to baseline (or initial) blood pressure, mean values of poststroke blood pressure, stroke subtypes, age, antihypertensive treatment, and other clinical features. RESULTS: The recurrence rate had a J-curve relation to poststroke diastolic blood pressure but not to poststroke systolic blood pressure and baseline diastolic and systolic blood pressures. The stroke recurrence rate was 3.8% per patient-year in 94 patients who had a poststroke diastolic blood pressure of 80 to 84 mm Hg, significantly lower than the rates of 9.2% per patient-year (P < .05) and 11.4% per patient-year (P < .01) in those with a lower and higher poststroke diastolic blood pressure, respectively. The range of poststroke diastolic blood pressure accompanying the lowest stroke recurrence rate was higher in patients with atherothrombotic (85 to 89 mm Hg) than in those with lacunar infarction (80 to 84 mm Hg). Neither antihypertensive therapy nor patients' age affected this phenomenon. CONCLUSIONS: The present study suggests that lower blood pressure does not always result in favorable effects on stroke recurrence. The effects of poststroke blood pressure and antihypertensive therapy on stroke recurrence may be complicated by the J-curve phenomenon. PMID- 8248967 TI - Clinical analysis of incidentally discovered unruptured aneurysms. AB - BACKGROUND AND PURPOSE: We analyzed the risk factors for rupture of an intracranial aneurysm based on a retrospective angiographic study of ruptured and unruptured aneurysms. METHODS: The 44 cases of asymptomatic aneurysms were selected from 1612 patients whose lesions had been discovered fortuitously by angiography (2.7%) during the period from 1980 to 1989. All these patients were free from any sign of intracranial aneurysm. The variations in age, sex, and location of the aneurysms were analyzed compared with 638 ruptured aneurysms that had been treated in our institute during the same period. The size, shape, and arterial geometry of the unruptured aneurysms were examined angiographically. RESULTS: Unruptured aneurysm was discovered fortuitously in 44 (2.7%) of 1612 patients, with greater incidence in women aged older than 60 years. Unruptured aneurysms were less likely to occur in the anterior communicating artery (12.8%) and the middle cerebral artery (6.4%). However, they were frequently found in the internal carotid artery, with an incidence of 10.6% in cavernous portion of the internal carotid artery, 19.1% in the internal carotid-ophthalmic artery, 19.1% in the internal carotid-posterior communicating artery, and 12.8% in the internal carotid-anterior choroidal artery. Seven of the nine internal carotid-posterior communicating artery aneurysms showed a hypoplastic or aplastic posterior communicating artery. The mean diameter of the unruptured aneurysms was 4.8 mm, and 80% were smaller than 6 mm. CONCLUSIONS: Intracranial aneurysms are formed not only at the bifurcation of an artery but also at its branching and bending points. However, an aneurysm located at the bifurcation, such as the anterior communicating artery and the middle cerebral artery, bleeds easily in contrast with lateral aneurysms such as those found at the branching and bending points on the internal carotid artery. PMID- 8248968 TI - Computer-assisted three-dimensional image analysis of cerebral amyloid angiopathy. AB - BACKGROUND AND PURPOSE: Microaneurysms and fibrinoid necrosis of cerebral cortical arteries have been reported to be related to the pathogenesis of intracerebral hemorrhage associated with cerebral amyloid angiopathy. To elucidate the pathogenesis of such vascular lesions, we conducted the present study. METHODS: Five hundred serial sections from brain tissue of a patient with severe amyloid angiopathy and intracerebral hemorrhage were analyzed histologically and immunohistochemically. Three-dimensional reconstructions of the vascular lesions were performed using a computer-assisted image analysis system. RESULTS: The microaneurysms were found to develop in small cortical arteries with diameters of about 40 to 50 microns. They were spindle-shaped dilatations, with a maximum diameter of about 200 microns, and appeared within vascular segments bearing severe amyloid deposition. In the walls of the aneurysms, the intima was thickened, and the media and adventitia showed thinning and disruption. Fibrinoid necrosis was found in the vascular walls of the most dilated, middle portions of the aneurysm. The vascular walls undergoing fibrinoid necrosis did not show any beta/A4 or cystatin C but presented with fibrinogen like immunoreactivities, indicating invasion of plasma components. CONCLUSIONS: These results suggested the following sequential events for the pathogenesis of the cerebral amyloid angiopathy-associated vascular lesions leading to hemorrhage: (1) damage of the media and adventitia due to severe amyloid deposition results in dilatation of the cortical arteries, (2) the vascular dilatation progresses and is accompanied by thickening of the intima and disruption of the media and adventitia (microaneurysm formation), (3) plasma components invade to the vascular wall (fibrinoid necrosis), and (4) finally, hemorrhage develops. PMID- 8248969 TI - Atrial septal aneurysm and patent foramen ovale as risk factors for cryptogenic stroke in patients less than 55 years of age. A study using transesophageal echocardiography. AB - BACKGROUND AND PURPOSE: An association between atrial septal aneurysm and embolic events has been suggested. Atrial septal aneurysm has been shown to be associated with patent foramen ovale and, in some reports, with mitral valve prolapse. These two latter cardiac disorders have been identified as potential risk factors for ischemic stroke. The aim of this prospective study was to assess the role of atrial septal aneurysm as an independent risk factor for stroke, especially for cryptogenic stroke. METHODS: We studied the prevalence of atrial septal aneurysm, patent foramen ovale, and mitral valve prolapse in 100 consecutive patients < 55 years of age with ischemic stroke who underwent extensive etiological investigations. We compared these results with those in a control group of 50 consecutive patients. The diagnosis of atrial septal aneurysm and patent foramen ovale relied on transesophageal echocardiography with a contrast study and that of mitral valve prolapse, on two-dimensional transthoracic echocardiography. RESULTS: Stepwise logistic regression analysis showed that atrial septal aneurysm (odds ratio, 4.3; 95% confidence interval, 1.3 to 14.6; P = .01) and patent foramen ovale (odds ratio, 3.9; 95% confidence interval, 1.5 to 10; P = .003) but not mitral valve prolapse were significantly associated with the diagnosis of cryptogenic stroke. The stroke odds of a patient with both atrial septal aneurysm and patent foramen ovale were 33.3 times (95% confidence interval, 4.1 to 270) the stroke odds of a patient with neither of these cardiac disorders. For a patient with atrial septal aneurysm of > 10-mm excursion, the stroke odds were approximately 8 times the stroke odds of a patient with atrial septal aneurysm of < 10 mm. CONCLUSIONS: This study shows that atrial septal aneurysm and patent foramen ovale are both significantly associated with cryptogenic stroke and that their association has a marked synergistic effect. Atrial septal aneurysms of > 10-mm excursion are associated with a higher risk of stroke. PMID- 8248970 TI - Lacunar infarcts and white matter attenuation. Ophthalmologic and microcirculatory aspects of the pathophysiology. AB - BACKGROUND AND PURPOSE: By means of neurological and ophthalmologic examinations we considered whether there is a microcirculatory disorder not related to hypertension and diabetes in patients with lacunar infarcts and whether there are microcirculatory differences in patients with lacunar infarcts compared with those with white matter attenuation. METHODS: Eighty neurological patients with a lacunar infarct underwent computed tomography and, based on the results, were prospectively assigned to subgroups as follows: (1) patients without changes; (2) patients with white matter attenuation but without lacunar infarcts; (3) patients with lacunar infarcts alone; and (4) patients with both lacunar infarcts and white matter attenuation. Clinical and ophthalmologic parameters were monitored. The retinal microcirculation was studied by videofluorescence angiography. These neurological patients were compared with control ophthalmologic patients matched for age, sex, hypertensive and diabetic ocular fundus changes, and smoking habits. RESULTS: On average, the 80 patients with lacunar infarcts had a significantly (P = .0001) slower arteriovenous passage time (2.6 +/- 0.7 seconds) than the ophthalmologic control subjects (1.6 +/- 0.6 seconds). Arteriovenous dye passage time through the retinal microcirculation was nearly normal (2.2 +/- 0.8 seconds) in patients with white matter attenuation alone, but was significantly prolonged in patients with lacunar infarcts (2.9 +/- 0.8 seconds, P = .00085) or both white matter attenuation and lacunar infarcts (2.8 +/- 0.4 seconds, P = .008). CONCLUSIONS: Patients with lacunar infarcts are characterized by an additional disorder of retinal microcirculation independent of arterial hypertension and diabetes. Our data suggested that white matter attenuation and lacunar infarcts may be phenomena with only weak interdependence. PMID- 8248971 TI - Cerebral venous thrombosis associated with pregnancy and puerperium. Review of 67 cases. AB - BACKGROUND AND PURPOSE: Cerebral venous thrombosis is characterized by its clinical pleomorphism and pathogenetic variability. We studied 67 patients with cerebral venous thrombosis associated with pregnancy and puerperium and compared them with 46 other cases unrelated to obstetric causes to disclose differences in their clinical presentation, neuroradiological findings, clinical course, and prognosis. METHODS: In this retrospective study, we analyzed the clinical, laboratory, and neuroimaging findings of 113 patients collected consecutively at our institute. The diagnosis of cerebral venous thrombosis was confirmed by angiography, magnetic resonance imaging, or neuropathological study. RESULTS: Patients with cerebral venous thrombosis associated with pregnancy and puerperium were younger (average age, 26 versus 36 years), and in most, the onset of symptoms was acute (82% versus 54%; P = .003). The evolution of symptoms reached a plateau within 10 days in 70% of patients with thrombosis from obstetric causes, compared with only 45% in those from other causes (P = .01). Anemia was more frequent in the obstetric group (64% versus 26%; P = .00001). There were no differences regarding neurological and neuroradiological findings. Although the initial severity of illness was similar in both groups, the final outcome was considered good in 80% of patients with obstetric causes, compared with 58% of patients with other causes (P = .01); mortality rates were 9% and 33%, respectively (P = .002). CONCLUSIONS: Cerebral venous thrombosis associated with pregnancy and puerperium has a more acute onset and a better prognosis than thrombosis due to other causes. These findings might be helpful in the diagnostic and therapeutic strategies for patients with cerebral venous thrombosis. PMID- 8248972 TI - Hemodilution improves cerebral hemodynamics in internal carotid artery occlusion. AB - BACKGROUND AND PURPOSE: Hemodilution may be a useful form of therapy for patients with carotid occlusive disease and hemodynamic ischemia. Accordingly, we evaluated the effects of hemodilution on cerebral hemodynamics and oxygen metabolism in patients with carotid artery occlusion. METHODS: Using positron emission tomography, we analyzed regional cerebral blood flow, oxygen extraction fraction, oxygen metabolic rate, and blood volume before and after isovolemic hemodilution in five patients with unilateral internal carotid artery occlusion and minor stroke. Hemodilution was accomplished by phlebotomy of 400 mL and infusion of 400 mL of hydroxyethyl starch. RESULTS: Before hemodilution, the patients had a significant decrease in blood flow and oxygen transport along with significantly elevated oxygen extraction fraction in the cerebral hemisphere with carotid artery occlusion compared with six control subjects. After hemodilution, the hematocrit and arterial oxygen content decreased from 41.2% and 18.6 mL/dL to 36.3% and 16.5 mL/dL, respectively. Both cerebral blood flow and oxygen transport were increased and oxygen extraction fraction was decreased without any change in oxygen consumption. The degree of increase in blood flow and oxygen transport was positively correlated with the ratio of oxygen extraction fraction to blood volume before hemodilution. CONCLUSIONS: These findings indicate that hemodilution improves oxygen transport as well as blood flow in patients with internal carotid occlusion and decreased perfusion and that this improvement may be more prominent in patients with a severely compromised hemodynamic state. Thus, hemodilution may be useful in patients with hemodynamic ischemia. PMID- 8248973 TI - Early temporal variation of cerebral metabolites after human stroke. A proton magnetic resonance spectroscopy study. AB - BACKGROUND AND PURPOSE: Proton magnetic resonance spectroscopy has documented declines in normal metabolites and long-term elevation of lactate signal after stroke in humans. Within days of stroke, leukocytes infiltrating the infarct zone may produce much of the lactate seen in the subacute and chronic periods. METHODS: We examined 10 patients by localized proton magnetic resonance spectroscopy with one-dimensional spectroscopic imaging within the first 60 hours after acute nonhemorrhagic cerebral infarction, a period before abundant leukocyte infiltration. Follow-up studies on day 8 to 17 after stroke were performed on 7 of these patients. RESULTS: Initially, the lactate magnetic resonance signal was elevated in all patients. The N-acetyl-aspartate peak within the lesion was reduced below contralateral normal brain in all but two. At subsequent examination, significant declines had occurred in lesion maximum lactate and N-acetyl-aspartate signals, with average changes of -36 +/- 11% per week and -29 +/- 9% per week, respectively. Declines in lesion creatine/phosphocreatine and in choline-containing compound peaks occurred in some patients but did not attain statistical significance for the group as a whole. Estimated lesion volume correlated positively with both total (r = .75, P = .012) and lesion maximum (r = .74, P = .015) lactate signal. CONCLUSIONS: Elevated lactate signal is reliably detectable by magnetic resonance spectroscopy after acute cerebral infarction in humans. Clearance of lactate occurs despite the potential contribution of lactate-producing leukocytes in the subacute stage. Delayed loss of N-acetyl-aspartate signal in second examinations suggests that late death of viable cells may occur within the first 2 weeks after cerebral infarction. PMID- 8248974 TI - Relationship of transcranial Doppler flow velocities and arteriovenous malformation feeding artery pressures. AB - BACKGROUND AND PURPOSE: Feeding mean arterial pressure immediately proximal to the nidus of arteriovenous malformations may influence the frequency of spontaneous intracerebral hemorrhage. This study assessed the usefulness of transcranial Doppler ultrasound velocities as a noninvasive estimate of feeding mean arterial pressure. METHODS: We studied 41 patients undergoing 73 staged treatments of arteriovenous malformations with endovascular embolization, surgery, or both. Before treatment during the awake state, transcranial Doppler mean and peak velocities were recorded in proximal Willisian vessels. During superselective angiography with the patient under conscious sedation or during surgery with the patient under general anesthesia, feeding mean arterial pressure was measured through a 1.5F transfemoral intracranial microcatheter or a 26-g needle by direct puncture. Measurement of insonated artery diameter was possible in 41 embolizations, and a flow velocity index (mL/min) and Reynolds' number were estimated. RESULTS: Mean +/- SEM feeding mean arterial pressure was 38 +/- 2 mm Hg at a systemic mean arterial pressure of 77 +/- 2 mm Hg; mean velocity was 102 +/- 4 cm/s. There was an inverse correlation between feeding mean arterial pressure and parent artery mean velocity (y = -0.74x + 130, r = .35, P = .0025). The best correlation was for the first treatment in each patient (n = 27) using the highest peak velocity obtainable in a Willisian vessel ipsilateral to the arteriovenous malformation (y = -1.61x + 221, r = .62, P = .0005). Flow velocity index (775 +/- 106 mL/min) did not correlate with feeding mean arterial pressure, but there was a weak correlation with Reynolds' number (y = -12x + 1616, r = .27, P = .1283). Mean Reynolds' number was 1257 +/- 119. CONCLUSIONS: Transcranial Doppler mean velocity is correlated with feeding mean arterial pressure but only weakly predictive. Considerations influencing the relation of distal feeding mean arterial pressure to proximal mean velocity might include the influence of other fistulae in the circuit between major inflow and outflow channels as well as turbulent flow at vascular branch points between point of insonation and the nidus, as suggested by Reynolds' number values of more than 400. PMID- 8248975 TI - Clinical features, pathogenesis, and computed tomographic characteristics of internal watershed infarction. AB - BACKGROUND AND PURPOSE: Infarction in the internal border-zone region has been described radiologically and pathologically. The aim of this study was to define the clinical and pathophysiological correlates of internal watershed infarction. METHODS: Eighteen consecutive stroke patients with evidence of internal watershed infarction on computed tomography (CT) were studied. RESULTS: Two CT patterns were identified: 6 patients had confluent internal watershed infarction (CIWI), and 12 patients had partial internal watershed infarction (PIWI). Syncopal symptoms and/or documented hypotension were prominent in both groups. Patients with CIWI usually presented with stepwise onset of contralateral hemiplegia and recovered poorly; patients with PIWI usually had discrete episodes of brachiofacial sensorimotor deficit and good recovery. Both groups had evidence of cortical involvement as part of their clinical deficit. Severe carotid occlusive disease was seen in 10 patients, and 12 patients had evidence of transiently impaired cardiac output. Carotid disease (P < .001), cardiac disease (P < .01), and diabetes mellitus (P < .01) were more prevalent in patients with internal watershed infarction compared with our stroke population as a whole. CONCLUSIONS: Distinguishing internal watershed infarction from lacunar and other subcortical infarctions is important because the different pathological mechanisms demand different therapeutic strategies. PMID- 8248976 TI - Functional image of dynamic computed tomography in diagnostic and prognostic evaluation of ischemic stroke within the first 6 hours. AB - BACKGROUND AND PURPOSE: It is important to make a diagnosis before a low-density area appears on computed tomography for appropriate management of acute ischemic stroke. We report the diagnostic and prognostic usefulness of functional image of dynamic computed tomography for acute ischemic stroke. METHODS: Forty-seven patients with ischemic strokes within 6 hours of ictus underwent dynamic computed tomography in which functional images were obtained. These findings were compared with angiographic findings, follow-up computed tomography, and clinical outcome. RESULTS: The functional images were categorized into three groups: (1) cortical type: abnormalities on time to peak image and/or corrected mean transit time image involving mainly cortical structures (29 cases); (2) noncortical type: abnormalities on either or both images limited to noncortical structures (7 cases); and (3) normal type: no abnormalities on both images (11 cases). Cortical type as a diagnostic test for arterial trunk occlusion had a good sensitivity (100%), specificity (95%), and accuracy (98%). Infarction volume on follow-up computed tomography correlated with extension of prolonged time-to-peak area (r = .80, P < .01) and that of prolonged corrected mean transit time area (r = .81, P < .01). Cortical type was associated with significantly unfavorable outcome (P < .01). CONCLUSIONS: Functional image of dynamic computed tomography findings predicted arterial trunk occlusion, infarction volume, and clinical outcome. Therefore, this technique would be useful not only for indicating definitive angiography and subsequent therapy but for evaluating the effectiveness of surgical or medical recanalization. PMID- 8248977 TI - Hematologic abnormalities occur in both cortical and lacunar infarction. AB - BACKGROUND AND PURPOSE: Primary hematologic abnormalities are a rare but established cause of ischemic stroke. In addition, activation of hemostatic parameters is often present during the acute phase of stroke. However, it is uncertain whether these abnormalities occur in both cortical and lacunar infarction; this study aimed to further assess this issue. METHODS: Hematologic parameters (prothrombin, activated partial thromboplastin, thrombin clotting, and euglobulin lysis times; and fibrinogen, fibrinopeptide A, antithrombin III, protein C, protein S, and plasminogen levels) were measured in 19 patients within 48 hours of the onset of acute cerebral infarction. These patients included 10 with cortical infarcts and 9 with lacunar infarcts, as determined by standard clinical and radiological criteria. RESULTS: Five patients with lacunar infarction and 7 patients with cortical infarction demonstrated raised fibrinopeptide A levels, indicating enhanced thrombin activity. Fibrinolysis, assessed by the euglobulin lysis time, was impaired in 6 of 9 patients with lacunar infarction and in 2 of 10 patients with cortical infarction. Lupus anticoagulants were detected in 3 patients with lacunar infarction and in 1 patient with cortical infarction. Three patients in each group displayed decreased antithrombin III function, and 1 patient with a lacunar infarction had a low protein C level. CONCLUSIONS: Primary hematologic disorders and secondary hemostatic derangements may occur in patients with either cortical or lacunar infarction. PMID- 8248978 TI - A quantitative investigation of the variability of the major cerebral arterial territories. AB - BACKGROUND AND PURPOSE: We have previously reported that the topographic variability of the territories of the anterior, middle, and posterior cerebral arteries is much larger than is generally considered in the literature. In the current study, we quantitatively investigated the variability of the territorial distribution of the major cerebral arteries and analyzed its relation to the variability of the circle of Willis. METHODS: In 23 human brains, the volumes of the major cerebral territories were calculated and the diameters of the contributing arteries were measured and standardized for size. RESULTS: The variability of the standardized territorial volumes proved to be considerably large and strongly correlated with the variability of the diameters of the arteries emanating from the circle of Willis. Furthermore, the relative vascular densities in the gray and white matter were determined. Taking these densities into account, an estimation of the relative peripheral resistance of the contributing arteries was made. CONCLUSIONS: We hypothesize that the morphological variability of the cerebral vascular system is related to the peripheral resistance of the major cerebral arteries and, consequently, to flow patterns, both of which are hemodynamic factors. We suggest that hemodynamic factors predominantly determine the form and size of the cerebral vascular system. PMID- 8248979 TI - Correlations between morphological changes in platelet aggregates and underlying endothelial damage in cerebral microcirculation of mice. AB - BACKGROUND AND PURPOSE: The aim of this study was to test the hypothesis that, once formed, platelet aggregates may injure underlying cerebrovascular endothelium. Such injury could make the same site selectively attractive to the next wave of passing emboli or activated platelets. This vicious circle could account for repetitive, stereotypic symptoms in transient ischemic attacks. METHODS: In pial arterioles, minor endothelial injury was produced by a laser/dye technique. After various periods of platelet aggregation at the damaged site, the vessels were fixed in situ for electron microscopic study. The degree of platelet activation (rounded and/or degranulated forms) was evaluated by counting these forms in the electron photomicrographs. These counts were related to the degree of endothelial damage ascertained in the micrographs. Other statistical relations were also examined. RESULTS: Endothelial damage progressed in parallel with the duration of platelet aggregation and the degree of platelet degranulation at the site. Correlations were number of activated platelets versus degree of damage, r = .43, P < .03; duration of aggregation versus damage, r = .52, P < .01; and number of degranulated platelets versus the degree of endothelial damage, r = .83, P < .001. If an aggregate embolized, endothelial damage did not appear to progress. No correlation existed between the duration of exposure to the laser and the degree of injury. CONCLUSIONS: The parallel between changes in platelets and endothelial damage could represent either an effect of endothelium on platelets or an effect of platelets on endothelium. Although the former alternative cannot be totally ruled out, the observations seem to fit best the hypothesis that progressive endothelial damage can result from increasing activation and degranulation of overlying platelets. PMID- 8248980 TI - Effect of phorbol myristate acetate on cerebral blood flow in normal and neutrophil-depleted rats. AB - BACKGROUND AND PURPOSE: Recent evidence suggests a possible role for leukocytes in ischemic brain injury. This study examined the effect of activation of endogenous circulating leukocytes on cerebral blood flow in normal and neutrophil depleted rats. METHODS: Leukocytes were activated by rapid injection of either 50 micrograms/kg phorbol 12-myristate 13-acetate, a protein kinase C activator, or an equimolar amount of the chemotactic peptide N-formyl-methionyl-leucyl phenylalanine, into the right carotid artery. Control rats received an equal volume of dimethyl sulfoxide in saline vehicle. H2-clearance cerebral blood flow was measured in each of the three groups and in vinblastine-treated, neutrophil depleted rats after carotid artery injection of phorbol. RESULTS: Phorbol 12 myristate 13-acetate dramatically decreased circulating leukocyte and platelet counts from 5 to 120 minutes after infusion and decreased regional cerebral blood flow in the ipsilateral parietal cortex from a baseline of 119 +/- 14 mL.min 1.100 g-1 (mean +/- SEM) to 49 +/- 5 mL.min-1.100 g-1 at 30 minutes (P < .05). Decreased flow persisted for the 2-hour study. Neither N-formyl-methionyl-leucyl phenylalanine or vehicle had an effect on cerebral blood flow. In the neutrophil depleted rats the initial decrease in cerebral blood flow at 30 and 60 minutes after infusion of phorbol was observed, but cerebral blood flow was restored to 70% to 80% of its baseline value (P > .05 versus baseline) by 90 to 120 minutes. CONCLUSIONS: The early phorbol 12-myristate 13-acetate-induced decrease in cerebral blood flow may be due to the effects of protein kinase C activation on vascular smooth muscle or on platelet aggregation, whereas the persistent decrease in cerebral blood flow appears to be mediated in part by neutrophil activation. PMID- 8248981 TI - The ultrastructure of photochemically induced thrombi with embolization in a rat model. AB - BACKGROUND AND PURPOSE: Photochemical techniques, currently used in stroke and cancer research, produce endothelial damage and thrombosis. To further characterize these thrombi and to determine whether they embolize, we studied the ultrastructure of photochemically damaged carotid arteries and small vessels distal to the irradiated carotid. METHODS: The right carotid artery of 9 Wistar rats was irradiated with a laser (632 nm, 200 mW/cm2, 15 minutes) after the injection of the photosensitizing dye Photofrin II, 12.5 mg/kg. There were 6 additional control rats: laser only, 2 rats; dye only, 2; carrier only (5% dextrose), 1; and normal, 1. The carotid artery and cerebral arterioles were studied using scanning and transmission electron microscopy. RESULTS: Endothelial damage was present in all irradiated carotid arteries, and consisted of exposure of the subendothelium and the formation of a nonocclusive thrombus. Although most cerebral arterioles were normal, 32 of these vessels contained peripheral blood elements, with platelet or red blood cell aggregates present in 15. The endothelium adjacent to the aggregates was intact. A few scattered endothelial cells had been lost in the carotid artery of control animals (compatible with normal cell turnover), with a few platelets adhering to the exposed subendothelium. CONCLUSIONS: Aggregates of blood cells and platelets in cerebral vessels in the absence of endothelial denudation verifies embolism as the mechanism for cerebral vascular occlusion in this experimental model. The possibility of embolization distal to the site of photochemical irradiation has implications for potential applications of photochemistry for cancer treatment and the ablation of vascular malformations and/or aneurysms. PMID- 8248982 TI - The kinetics of lymphocyte subsets and macrophages in subarachnoid space after subarachnoid hemorrhage in rats. AB - BACKGROUND AND PURPOSE: Although it has been suggested that humoral immunity plays a role in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage, there has been no quantitative assay for cellular immunity. We studied the kinetics of immune cells in the subarachnoid space after subarachnoid hemorrhage in the rat. METHODS: One hundred fourteen Sprague-Dawley rats were used in this study. The animals were divided into two groups and injected with either autologous blood (0.3 mL) or saline into the major cistern. They were killed at the specified time: 10 minutes or 1, 2, 3, 5, or 7 days after subarachnoid hemorrhage. For immunohistochemical analysis, the rats' whole brains were frozen, and cryostat sections were prepared. For flow cytometric analysis of immune cell presence, their whole brains underwent enzymatic digestion. RESULTS: Histopathologic study revealed pathological change of the arterial wall, and immunohistochemical study revealed the existence of macrophages and T cells in the subarachnoid space in animals with a survival time of 2 to 5 days after subarachnoid hemorrhage. A flow cytometric study revealed the peak of appearance of T cells and macrophages 2 days after subarachnoid hemorrhage. The helper suppressor T cell ratio also reached a peak 2 days after subarachnoid hemorrhage. CONCLUSIONS: A serial response of immunoreactive cells, which resembles that of the chronic allergic reaction observed in autoimmune diseases or delayed-type hypersensitivity, exists in the subarachnoid space after subarachnoid hemorrhage. The present results suggest that the initial response in cellular immunity, which is followed by humoral immunity and eicosanoid reactions, plays a role in eliciting the development of cerebral vasospasm. PMID- 8248983 TI - Evidence supporting a role for programmed cell death in focal cerebral ischemia in rats. AB - BACKGROUND AND PURPOSE: Cells die by one of two mechanisms, necrosis or programmed cell death. Necrosis has been implicated in stroke and occurs when the cytoplasmic membrane is compromised. Programmed cell death requires protein synthesis and often involves endonucleolytic cleavage of the cellular DNA. We assessed the potential contribution of programmed cell death to ischemia-induced neuronal death. METHODS: Cycloheximide (protein synthesis inhibitor; 1 mg/kg per 24 hours) or vehicle (1 mL/kg per 24 hours) was continuously infused into the right cerebral ventricle of spontaneously hypertensive rats. Neocortical focal ischemia was produced by tandem occlusion of the right common carotid artery and the ipsilateral middle cerebral artery. After 24 hours the brain was stained with 2% 2,3,5-triphenyltetrazolium and the ischemic zone quantitated. Protein synthesis was determined by [3H]methionine incorporation into acid-precipitated protein. DNA integrity was determined in isolated DNA by gel electrophoresis and in whole cells by flow cytometry. RESULTS: Continuous cycloheximide infusion caused approximately 70% reduction in cortical protein synthesis. Cycloheximide also reduced the size of the infarction produced by focal cerebral ischemia when compared with controls (ischemic brain volume, 147.5 +/- 25.9 and 188.7 +/- 16.8 mm3 for cycloheximide and saline, respectively; P < .01), suggesting that protein synthesis may contribute to cell death. Purified DNA from the ischemic zone showed evidence of endonucleolytic degradation when fractionated by gel electrophoresis. Flow cytometric analysis demonstrated increased propidium iodide fluorescence in intact cells isolated from ischemic cortex, indicating an increased accessibility of degraded DNA to the intercalating dye. CONCLUSIONS: New protein synthesis appears to contribute to ischemic cell death in which endonucleolytic DNA degradation is apparent. These observations implicate programmed cell death in ischemic injury and may open unique therapeutic approaches for the preservation of neurons in stroke. PMID- 8248984 TI - Nitric oxide and S-nitroso-L-cysteine as endothelium-derived relaxing factors from acetylcholine in cerebral vessels in cats. AB - BACKGROUND AND PURPOSE: The predominant view is that the endothelium-derived relaxing factor generated by acetylcholine from blood vessels is nitric oxide. However, there is evidence suggesting that certain nitric oxide-containing compounds such as nitrosothiols resemble the endothelium-derived relaxing factor generated by acetylcholine more closely than does nitric oxide itself. Accordingly, we compared the effects of nitric oxide and S-nitroso-L-cysteine on cerebral arteriolar caliber in relation to the associated increments in nitrite concentration in the effluent. METHODS: Acetylcholine, nitric oxide, and S nitroso-L-cysteine were administered by continuous superfusion in oxygen-free solution through the space under a cranial window in anesthetized cats. Nitrite concentration was measured in the effluent. The degree of vasodilation induced was evaluated in relation to the increment in nitrite concentration. RESULTS: All agents induced dose-dependent vasodilation and dose-dependent increments in nitrite concentration in the effluent. For any given degree of vasodilation, the increments in nitrite concentration were equivalent during acetylcholine or S nitroso-L-cysteine infusion, whereas the nitrite concentrations were 10 times higher during nitric oxide infusion. After administration of nitroarginine, a competitive inhibitor of nitric oxide synthesis from arginine, there was depression in the vasodilation as well as the increment in nitrite concentration induced by acetylcholine. CONCLUSIONS: S-Nitroso-L-cysteine resembles endothelium derived relaxing factor from acetylcholine more closely than does nitric oxide. PMID- 8248985 TI - Acidic foci within the ischemic penumbra of the New Zealand white rabbit. AB - BACKGROUND AND PURPOSE: In vivo panoramic imaging of reduced nicotinamide adenine dinucleotide (NADH), intracellular brain pH (pHi), and cortical blood flow was used to characterize the ischemic penumbra during focal ischemia. During global ischemia, hypoxia, and status epilepticus, the development of cortical acidic foci has been observed. The hypothesis tested was that during focal ischemia, acidic foci develop, which may lead to recruitment of the ischemic penumbra into infarction. METHODS: Five fasted New Zealand White rabbits underwent middle cerebral artery (MCA) occlusion under 1.5% halothane anesthesia through a retro orbital approach, and five animals served as controls. Brain pHi and cerebral blood flow were measured with in vivo umbelliferone fluorescence. RESULTS: Baseline brain pHi was 6.98 +/- 0.05, whereas cortical blood flow and NADH fluorescence measured 52.2 +/- 8.7 mL/100 g per minute and 35.5 +/- 3.7 gray scale units, respectively. Fifteen minutes after MCA occlusion, overall brain pHi and cortical blood flow of the ischemic penumbra measured 6.61 +/- 0.06 and 31.9 +/- 9.2 mL/100 g per minute. Over 3 hours there was normalization of pHi in the majority of the penumbra due to increases in cortical blood flow. Within the ischemic penumbra acidic foci developed with an initial pHi of 6.35 +/- 0.09 and cortical blood flow of 18.0 +/- 5.7 mL/100 g per minute. These foci remained acidic with increased NADH fluorescence despite being surrounded by cortex that was recovering from ischemia. On light microscopy, these acidic foci had a mixed pattern of neuronal injury. CONCLUSIONS: Within the ischemic penumbra, acidic foci develop that do not follow a vascular distribution and have microscopic evidence of ischemic neuronal injury. This suggests that there is a cortical selective vulnerability regarding pHi regulation and these acidic foci may lead to recruitment of the ischemic penumbra into infarction. PMID- 8248986 TI - Comparative effects of angiotensin-(1-7) and angiotensin II on piglet pial arterioles. AB - BACKGROUND AND PURPOSE: Recent investigations indicated that degradation fragments of angiotensins could be involved in the regulation of the cerebral circulation and that their effects might be mediated by prostaglandins. The present study was designed to examine the effect of angiotensin-(1-7), a major endogenous heptapeptide fragment, on cerebral arteriolar diameter and compare it with the octapeptide angiotensin II, and further to determine whether prostaglandins mediate their effects. METHODS: Newborn, anesthetized pigs were equipped with a closed cranial window, and the diameter of one pial arteriole was measured using intravital microscopy. RESULTS: Topical application of angiotensin (1-7) (n = 9) increased the diameter by 6.8 +/- 5.3% (mean +/- SEM), 10.4 +/- 5.2%, 14.3 +/- 5.9%, and 17.5 +/- 7.7% (P < .05) at 10(-7), 10(-6), 10(-5), and 10(-4) mol/L, respectively (baseline, 94 +/- 3 microns). Topical application of angiotensin II (n = 8) increased the diameter by 9.6 +/- 7.0%, 9.6 +/- 7.6%, 11.3 +/- 8.4% (P < .05), and 5.5 +/- 7.9% at 10(-7), 10(-6), 10(-5), and 10(-4) mol/L, respectively (baseline, 94 +/- 5 microns). After administration of indomethacin (5 mg/kg IV), which did not significantly change the baseline arteriolar diameter, neither angiotensin-(1-7) at 10(-4) mol/L nor angiotensin II at 10(-5) mol/L caused significant vasodilation. CONCLUSIONS: The results indicate that angiotensin-(1-7) is a modest dilator in the cerebral circulation, as is angiotensin II, and that prostaglandins may mediate responses. PMID- 8248987 TI - In vivo mapping of brain benzodiazepine receptor changes by positron emission tomography after focal ischemia in the anesthetized baboon. AB - BACKGROUND AND PURPOSE: Recent reports have shown an increase in specific binding (in vitro) of [3H]PK 11195 to peripheral-type benzodiazepine receptors in both experimental animals and humans, reflecting a glial/macrophagic reaction within and around focal ischemic insults. We have evaluated by positron emission tomography the time course of changes in brain uptake in vivo of 11C-labeled PK 11195 and flumazenil (an antagonist of central benzodiazepine receptors) as indirect and direct markers of neuronal loss, respectively, after focal cerebral ischemia. METHODS: Ten anesthetized baboons were submitted to sequential positron emission tomography studies between day 1 and day 91 after unilateral middle cerebral artery occlusion. The studies consisted of successive assessments, in the same positron emission tomography session, of [11C]PK 11195, [11C]flumazenil, cerebral blood flow, and oxygen consumption; late computed tomographic scans were obtained to map the approximate contours of infarction and to define a concentric peri-infarct area. RESULTS: We found a significant time-dependent increase in [11C]PK 11195 uptake in the peri-infarcted area, maximum at 20 to 40 days after occlusion. In contrast, there was a time- and perfusion-independent significant decrease in [11C]flumazenil uptake in the infarcted area, stable from day 2 onward, and already present in one baboon at day 1. Challenge studies with saturating doses of cold ligands confirmed that these changes represented alterations in specific binding. [11C]Flumazenil uptake was not affected in hypometabolic (but apparently noninfarcted, ie, deafferented) cortical areas. CONCLUSIONS: The delayed and apparently transient increases in [11C]PK 11195 specific uptake in vivo presumably represent glial/macrophage reaction; the marked depression in [11C]flumazenil specific binding, which appears selective for synaptic damage, is both precocious and sustained and thus may be better suited for the early assessment of ischemic damage in humans. PMID- 8248988 TI - Effect of a novel thromboxane A2 receptor antagonist, S-1452, on postischemic brain injury in rats. AB - BACKGROUND AND PURPOSE: Arachidonate metabolites have been implicated in the development of cerebral injury after ischemia. Particular importance has been placed on the balance of thromboxane A2 and prostaglandin I2 because of its regulative activity on platelet functions and arterial tone. The purpose of the present study was to shed light on the role of thromboxane A2 in postischemic brain injury. METHODS: We evaluated the effects of S-1452, a novel thromboxane A2 receptor antagonist, on brain edema, infarct areas, and survival rate in rats with middle cerebral artery occlusion. A transient middle cerebral artery occlusion model was produced by inserting a piece of silicon-coated nylon thread into the internal carotid artery. RESULTS: The ratio of plasma thromboxane B2 to 6-keto-prostaglandin F1 alpha significantly rose at 0 hour (P < .05), 1 hour (P < .01), 3 hours (P < .05), and 12 hours (P < .05) and then nearly returned to the normal level at 24 hours after reperfusion following 1-hour occlusion. Pretreatment with S-1452 (5, 10, or 50 mg/kg PO) significantly attenuated the increase in postischemic water content in the cerebral cortex perfused by the anterior cerebral artery and the cerebral cortex perfused by the middle cerebral artery in a dose-dependent manner but slightly attenuated it in the caudate putamen 24 hours after reperfusion following 1-hour occlusion. Pretreatment with S-1452 (10 mg/kg PO) also significantly decreased the areas of infarction in the front parts of the cerebrum. The survival rate of animals after 2 hours of occlusion tended to be improved by treatment with S-1452 (10 mg.kg-1.d-1 PO), although there was no statistical significance. CONCLUSIONS: Our results suggest that thromboxane A2 is closely related to postischemic brain injury in the early phase of recirculation and that S-1452 may have a protective effect on postischemic brain injury. PMID- 8248989 TI - Effect of cerebral blood flow generated during cardiopulmonary resuscitation in dogs on maintenance versus recovery of ATP and pH. AB - BACKGROUND AND PURPOSE: Cardiopulmonary resuscitation with external chest compression generates low perfusion pressures that may be inadequate for restoring cerebral metabolism and may worsen intracellular pH. We tested the hypothesis that cerebral reperfusion with a low perfusion pressure after arrest restores brain adenosine triphosphate (ATP) and pH to levels attained at the same perfusion pressure without preceding complete ischemia. METHODS: Brain ATP and intracellular pH were measured by magnetic resonance spectroscopy, and cerebral blood flow was measured with microspheres in anesthetized dogs. External chest compressions were begun in group A (n = 6) immediately after the onset of arrest (ie, arrest time zero) and in group B (n = 10) after 6 minutes of arrest (ie, arrest time 6 minutes). In both groups, mean cerebral perfusion pressure was regulated at 30 mm Hg for 70 minutes by adjustment of inflation pressure of a pneumatic thoracic vest. RESULTS: At 12 minutes of resuscitation, cerebral blood flow was 27 +/- 4 mL/min per 100 g in group A and 21 +/- 4 mL/min per 100 g in group B, but ATP in group B (58 +/- 10% of prearrest) was less than in group A (105 +/- 6%). With prolonged resuscitation, ATP deteriorated to near zero levels in dogs in group B, with blood flow less than 15 mL/min per 100 g. Dogs with greater blood flow never achieved complete metabolic recovery. In group B, intracellular pH was unchanged from the 6.3 value at the start of resuscitation, even in those dogs with extremely low blood flows. CONCLUSIONS: Levels of cerebral perfusion pressure sufficient to maintain cerebral oxidative metabolism without complete ischemia during cardiopulmonary resuscitation are not sufficient to restore metabolism after complete ischemia during cardiopulmonary resuscitation. However, low "trickle" blood flow did not worsen intracellular acidosis. PMID- 8248990 TI - Diffusion and perfusion magnetic resonance imaging studies to evaluate a noncompetitive N-methyl-D-aspartate antagonist and reperfusion in experimental stroke in rats. AB - BACKGROUND AND PURPOSE: Diffusion magnetic resonance imaging (MRI) can quantitatively detect focal ischemic injury within minutes of onset, and perfusion MRI can evaluate the brain's microcirculation. N-Methyl-D-aspartate (NMDA) antagonists and reperfusion can reduce lesion size in stroke models. We used diffusion and perfusion MRI to evaluate the in vivo effects of a noncompetitive NMDA antagonist, CNS 1102, in a temporary ischemia model. METHODS: Sixteen Sprague-Dawley rats underwent suture occlusion of the middle cerebral artery. Fifteen minutes after occlusion, animals were randomly assigned to treatment with CNS 1102 (n = 10) or placebo (n = 6), receiving a bolus of 1.13 mg/kg at that time and an infusion of 0.785 mg.kg-1.h-1 for the next 165 minutes. The placebo group received a saline bolus and infusion. Diffusion MRI studies by a spin-echo technique were initiated 30 minutes after occlusion and repeated every 30 minutes for the next 3 hours. Perfusion MRI studies were obtained using echo-planar imaging after injection of superparamagnetic iron oxide particles, immediately before and 15 minutes after withdrawal of the occluder at 3 hours after middle cerebral artery occlusion. At 24 hours, the animals were clinically evaluated (scale of 0 to 5) and electively killed, and the brain was stained with triphenyltetrazolium chloride to evaluate infarct size. RESULTS: Diffusion imaging demonstrated markedly reduced ischemic lesion area in the CNS 1102 group during occlusion--10.5 +/- 7.3% (mean +/- SEM) of the ischemic hemisphere (optic chiasm slice) at 30 minutes after occlusion versus 50.0 +/- 2.7% of the hemisphere in controls (P < .02). With reperfusion after 3 hours of temporary ischemia, diffusion imaging documented an additional 29% reduction of the ischemic lesion area in the CNS 1102-treated group (P < .01) compared with the prereperfusion ischemic lesion area, with no change in the placebo group. During occlusion, perfusion imaging demonstrated a relative signal intensity decline of 31.5 +/- 7.7% in controls and 83.4 +/- 7.6% in the CNS 1102 group (P < .005), indicating better perfusion in the latter group. After removal of the occluder, perfusion improved in both groups and was not significantly different. Post mortem infarct volume was 53.8 +/- 20.0 mm3 in the CNS 1102 group and 216.8 +/- 16.1 mm3 in the controls (P < .0001). Clinical outcome at 24 hours was 1.1 +/- 0.4 in the CNS 1102 group and 4.0 +/- 0.5 (scale of 0 to 5) in the controls (P < .005). CONCLUSIONS: This study demonstrates that CNS 1102 reduces early postischemic injury as documented by diffusion MRI and improves perfusion as documented by perfusion MRI and that reperfusion confers additional reduction of ischemic lesion size. PMID- 8248991 TI - Reversed alterations of hippocampal parvalbumin and protein kinase C-gamma immunoreactivity after stroke in spontaneously hypertensive stroke-prone rats. AB - BACKGROUND AND PURPOSE: Aging spontaneously hypertensive stroke-prone rats (SHR SP) were previously shown to develop neocortical strokes. Because the hippocampal CA1 is selectively vulnerable to abnormal brain perfusion, the neuropathological effects of spontaneous strokes were investigated on specific neurochemical alterations in two major cell types of the hippocampal CA1 in SHR-SP. METHODS: The immunoreactivity for the gamma-isoform of protein kinase C (in pyramidal cells) and parvalbumin (in interneurons) was determined in the hippocampal CA1 by applying monoclonal antibodies. Because chronic treatment with the calcium antagonist nimodipine prevents the development of strokes in SHR-SP, we compared SHR-SP (stroke) with age-matched nimodipine-treated rats (nonstroke). RESULTS: After stroke in control animals, we observed a strikingly enhanced immunoreactivity for protein kinase C-gamma in CA1 pyramidal cells compared with nimodipine-treated rats, which can be interpreted as the result of an increased activation of these cells. The pathological increase of protein kinase C-gamma immunoreactivity was accompanied by a reduced parvalbuminergic innervation of these pyramidal cells in symptomatic SHR-SP. CONCLUSIONS: Because parvalbumin is present in a subset of GABAergic inhibitory interneurons, these data suggest that increased activity of CA1 pyramidal cells after spontaneous stroke may partially be related to a decreased inhibitory input on these cells. PMID- 8248992 TI - Anatomic variation of the middle cerebral artery in the Sprague-Dawley rat. AB - BACKGROUND AND PURPOSE: The aim of this study was to record and analyze the anatomy of the middle cerebral artery in the rat in an attempt to identify a potential source of variation of quantitative infarct measurements in commonly used models of focal cerebral ischemia. METHODS: Over a period of 22 months, as part of ongoing studies, we routinely induced focal cerebral ischemia in 263 Sprague-Dawley rats. This was achieved by permanent occlusion of a length of the right middle cerebral artery approximately 1 to 5 mm distal to the point at which it traverses the rhinal fissure combined with 60 minutes of bilateral common carotid occlusion. The branching pattern of the middle cerebral artery was recorded for each animal by drawing and photography before occlusion and at the time of death. RESULTS: Six distinct branching patterns were observed. The majority (82.9%) of middle cerebral arteries studied conformed to the typical bifurcating vessel commonly referred to in the literature. However, the remaining 17.1% were far more variable with major, atypical branching, introducing, we believe, unnecessary and avoidable variation into quantitative measurements of ischemia in this and similar models. CONCLUSIONS: Middle cerebral artery anatomy should be routinely recorded in models of focal cerebral ischemia in which this artery is occluded distal to the olfactory tract, thus allowing improved analysis of data. PMID- 8248993 TI - Hemibody tremor related to stroke. AB - BACKGROUND: Hemibody tremor is an uncommon manifestation of stroke. We describe a case investigated by both brain magnetic resonance imaging and positron emission tomography using [18F]fluorodeoxyglucose. CASE DESCRIPTION: Three months after a pure motor stroke, a 65-year-old man developed a right arm and leg tremor. The tremor was of large amplitude, intermittent at rest; its frequency was 5 to 6 Hz. Neither rigidity nor akinesia was detected, and administration of L-dopa was ineffective. Brain magnetic resonance imaging revealed an ischemic lesion in the left centrum semiovale and a left caudate lacunar infarction. We suspected that the resting unilateral tremor was related to this lacunar lesion. Positron emission tomography demonstrated glucose hypermetabolism in the left sensorimotor cortex. CONCLUSIONS: This case suggests that unilateral tremor may be related to a lacunar stroke in the caudate nucleus and may be accompanied by an increased glucose metabolism in the contralateral sensorimotor cortex. PMID- 8248994 TI - Accelerated brain infarction in hypertension complicated by hereditary heterozygous protein C deficiency. AB - BACKGROUND: Protein C deficiency leads to reduced inhibition of coagulation and an increased likelihood of thrombosis. It is widely accepted that the most common syndromes associated with protein C deficiency are venous thrombosis and pulmonary thromboembolism, whereas arterial thrombosis is rare. Here we describe two patients with hypertension and hereditary heterozygous protein C deficiency who developed multiple lacunar infarcts. CASE DESCRIPTIONS: Patient 1 was a 46 year-old man with a history of hypertension who developed a right upper quadrantanopia and gradually progressive intellectual and behavioral deterioration. Patient 2 was a 61-year-old man with history of hypertension and two episodes of right-sided motor weakness who developed left sixth and seventh cranial-nerve palsies and reduced pinprick sensation in the right extremities. In both patients, magnetic resonance imaging revealed multiple small lesions in the pons as well as the bilateral basal ganglia, thalamus, corona radiata, and other subcortical structures, which are consistent with lacunar infarcts. Protein C activity and antigen levels were reduced to approximately one half of normal in these two patients, as well as in some of their family members who had no other serological or coagulation abnormalities. A diagnosis of heterozygous protein C deficiency type 1 was thus established. CONCLUSIONS: Although it remains uncertain whether protein C deficiency itself increases the risk of cerebral artery thrombosis, it may predispose a patient to develop multiple brain infarctions in association with hypertension. PMID- 8248995 TI - Congenitally abnormal plasminogen in juvenile ischemic cerebrovascular disease. AB - BACKGROUND AND PURPOSE: Congenitally abnormal plasminogen is characterized by markedly decreased fibrinolytic activity and has been reported mainly in association with venous occlusive disease. CASE DESCRIPTION: We found three young adult patients (34, 45, and 27 years old at onset) with ischemic cerebrovascular disease, all of whom had congenital plasminogen abnormalities but no other known risk factors. Hemostatic tests of all three patients revealed plasma plasminogen activities at almost one half of the normal level despite normal plasma plasminogen antigen levels. They were found to be heterozygotes with abnormal plasminogen (normal Ala-601[GCT] to abnormal Thr-601[ACT]) by DNA sequence analysis after polymerase chain reaction. CONCLUSIONS: Congenital plasminogen abnormalities could be one of the risk factors of juvenile ischemic cerebrovascular disease of the arterial as well as venous type. PMID- 8248996 TI - Postpartum cerebral angiopathy. Is there a role for sympathomimetic drugs? AB - BACKGROUND: Postpartum cerebral angiopathy is associated with the use of of ergot alkaloids. The exact mechanism is unclear but may be related to their sympathomimetic properties, as evidenced in patients already on other ergot derivatives who deteriorated only after taking additional sympathomimetic drugs. We postulate that sympathomimetic agents, independent of ergot alkaloids, may produce the same complication. CASE DESCRIPTION: A postpartum patient, initially presenting with headaches, subsequently manifested rapid neurological deterioration after ingesting isometheptene, a sympathomimetic drug. She was not on any ergot derivative but presented similar clinical and radiological manifestations. She experienced increased headache severity, visual disturbance, and seizures associated with multiple segmental cerebral vasoconstriction on angiography and increased T2-weighted signal in the occipital areas on magnetic resonance imaging. CONCLUSIONS: This case is additional evidence that sympathomimetic actions of some drugs, such as ergot derivatives and isometheptene, may lead to postpartum cerebral angiopathy. Documentation of medication used by postpartum women suffering similar complications is needed to verify these findings. PMID- 8248997 TI - Persistent primitive proatlantal intersegmental artery (proatlantal artery I) results in 'top of the basilar' syndrome. AB - BACKGROUND: Persistent primitive proatlantal intersegmental artery (proatlantal artery I) is an anastomotic channel between the carotid and vertebrobasilar arterial systems. Persistence of this embryonic anastomosis is very rare. The "top of the basilar" syndrome is usually due to occlusion of thalamoperforating arteries by emboli that rise from the vertebrobasilar arterial system. In this case, however, the emboli stemmed from atherosclerotic plaques in the carotid system. CASE DESCRIPTION: A 55-year-old woman was hospitalized with top of the basilar syndrome. A brain scan showed hypodense lesions extending from the mesencephalon to both thalami. Digital subtraction angiography of the cerebral circulation was performed, which revealed a proatlantal artery I on the left side. In addition, atheromatous plaques were seen along the internal and common carotid arteries on the same side. CONCLUSIONS: The occurrence of proatlantal artery I is extremely rare. Top of the basilar syndrome is also rarely encountered. To our knowledge, this is the first report of a patient with both conditions. PMID- 8248998 TI - Discrete cortical infarction with prominent impairment of thumb flexion. AB - BACKGROUND: Intracortical efferent zones in the primary motor cortex for thumb movements, namely thumb flexion, extension, adduction, and abduction, have been described in Macaca mulatta monkeys but not in humans. Even recent cortical mapping based on intraoperative monitoring does not provide information about intracortical efferent zones as it is not ethically possible to search the human motor cortex extensively by punctate electrodes. CASE DESCRIPTION: A 78-year-old man with discrete cortical embolism over the left central sulcus is described. Only a mild weakness of his right thumb was observed. Thumb flexion was affected to a greater degree than thumb extension, adduction, and abduction. The lesion ran along the bottom of the central sulcus and affected part of the precentral and postcentral gyri on both sides of it. Animal studies in Macaca mulatta monkeys have shown that the intracortical efferent zones for thumb movement, especially for thumb flexion, are located in the part of the motor cortex just adjoining the central sulcus that folds down from the "bank" to the depth of the central sulcus. It was possible to explain the prominent weakness of thumb flexion if we applied the results of the animal studies. CONCLUSIONS: Our case suggests that the same arrangement of intracortical efferent zones found in monkeys may also exist in human beings. PMID- 8248999 TI - Altered gene expression in cerebral ischemia. AB - BACKGROUND: Using the techniques of molecular biology, recent experimental studies have shown that cerebral ischemia induces a variety of changes in gene expression in the brain. SUMMARY OF REVIEW: During the early postischemic stages, protein synthesis in the brain is generally suppressed, but specific genes are expressed and their corresponding proteins may be synthesized, such as immediate early gene products (c-fos, c-jun, and zinc finger gene), heat-shock proteins, and amyloid precursor protein. The ability of neurons to induce such stress responses, which depends on both the severity of ischemia and the intrinsic nature of the neuronal populations, may be directly associated with neuronal death and survival after ischemia. Nerve growth factor and fibroblast growth factor are also induced after ischemia and may be related to repair processes, in which a role of glial cells is suggested. Postischemic events that may be associated with the altered gene expression include (1) induction of tolerance to ischemia after pretreatment with sublethal ischemia, (2) slow, progressive neuronal changes and the development of neuronal plasticity after ischemia, and (3) delayed neuronal changes in remote areas outside the cerebral ischemic focus. CONCLUSIONS: Because a variety of harmful stresses, including ischemia, elicit the same stress response and because this response is induced when total protein synthesis in the brain is nearly completely suppressed, this response may be vital to cell survival and repair. A successful induction of this response may induce resistance and survival of neurons after ischemia. However, failure or abortion of the response and persistent stresses may lead to neuronal death and possibly long-term changes and degeneration. PMID- 8249000 TI - The use of expected value as an aid to decisions regarding anticoagulation in patients with atrial fibrillation. AB - BACKGROUND: The method described provides a rational means for determining whether to institute chronic anticoagulation to prevent stroke in patients with chronic atrial fibrillation under a variety of clinical circumstances. SUMMARY OF COMMENT: The concept of expected value is used in conjunction with data from clinical studies to define the net value of anticoagulation to the patient. A full year of anticoagulation is warranted in patients with recent stroke or transient ischemic attack thought to be due to cardiogenic embolism who feel that stroke is a very serious event with nearly as much disvalue as death. If stroke has a lesser degree of negative value to the patient, or it is uncertain whether the stroke was in a large-vessel distribution, or it is uncertain whether a large vessel distribution stroke was due to cardiogenic embolism, 6 months or less of anticoagulation may be warranted. Indefinite anticoagulation is justifiable in most patients with chronic atrial fibrillation without a history of stroke or transient ischemic attack but may be contraindicated in certain patients at extremely low risk for embolism and in patients who place a low value on stroke relative to death and who have a modest increase in risk for fatal hemorrhage. CONCLUSIONS: The method described provides a means readily usable by clinicians to make anticoagulation decisions in patients with chronic atrial fibrillation that will address risk-benefit tradeoffs with somewhat greater precision than current approaches. PMID- 8249002 TI - Spinal infarction follow-up study. PMID- 8249001 TI - The Stroke Prevention Patient Outcomes Research Team. Goals and methods. AB - BACKGROUND AND PURPOSE: The aim of the present study, based at Duke University and involving 14 other institutions, is to identify the most appropriate and cost effective clinical strategies for prevention of ischemic (thrombotic or embolic) stroke in high-risk individuals and to design and test an intervention to disseminate this information to providers and the public. METHODS: The study uses (1) secondary data from literature review, Medicare claims, and population-based data from three epidemiological studies and (2) primary data generated in national physician and patient surveys and in demonstration trials. Phases I through III involve data collection and analysis using a decision/cost effectiveness model and consensus development methods. Phase IV includes intervention in physicians' practice patterns. Data is collected by literature survey and abstraction, review of medical records, claims analysis, and patient and physician surveys. CONCLUSIONS: A structured decision model and a well defined clinical focus provide a successful organization for a PORT on stroke prevention. PMID- 8249003 TI - The clinical problem of brain and heart. AB - Observations in an intensive care stroke unit showed that patients recovering from stroke sometimes perish unexpectedly from cardiac complications or sudden death. Compared with control subjects, stroke patients more often have cardiac arrhythmias, raised cardiac enzymes, and elevated plasma norepinephrine, suggesting a hypersympathetic state. We have found that involvement of the insula by experimental cerebral infarction is crucial in mediating the cardiac complications. Age and right as opposed to left hemisphere involvement represent additional risks for the cardiac complications of experimental stroke. These experimental observations now need to be tested clinically. PMID- 8249004 TI - Treatment of deep spontaneous intracerebral hematomas. A review. PMID- 8249005 TI - Mechanisms of hemostasis. Effect on intracerebral hemorrhage. AB - Aspects of the mechanisms of hemostasis are reviewed with particular emphasis on the assembly of enzyme complexes on the membranes of cells. The way this membrane fixation of reactions accelerates coagulation, inhibition, and fibrinolytic systems is discussed. The complex nature of the hemostatic system and the multiple complications of its abnormalities present a difficult problem for the neurologist and neurosurgeon managing the patient with intracerebral hemorrhage. This is a situation in which close consultation and collaboration among the neurological and hematologic specialists are imperative to achieve the most favorable outcome for the patient with this life-threatening situation. PMID- 8249006 TI - Mechanisms of ischemic brain damage with intracerebral hemorrhage. AB - The results of surgical evacuation of spontaneous intracerebral hematomas are disappointing. This is largely because experimental studies have now confirmed that the brain surrounding an intracerebral hematoma develops profound and extensive ischemia. The volume of this ischemic brain may exceed the volume of the hemorrhage several times. This has been demonstrated experimentally using 14C iodoantipyrene autoradiography in various modifications of the intracerebral hemorrhage model. These models have demonstrated that the pathophysiology of the ischemia is partly due to direct mechanical compression. There is also a component of the ischemic process induced by vasoconstrictor substances in blood. The diffuse uncontained type of hemorrhage (subarachnoid or intraventricular) causes a global reduction in cerebral perfusion pressure. The focal ischemic event is initiated at the time of hemorrhage and is largely irreversible. The experimental evidence to date indicates that neuroprotective agents (calcium channel blockers and N-methyl-D-aspartate receptor antagonists) reduce ischemic brain damage. Similarly, in immunosuppressed animals the amount of brain edema that follows the initial ischemic insult was reduced. These studies indicate that pharmacological neuroprotective strategies can minimize the brain damage that follows intracerebral hemorrhage. Early removal of the mass lesion may play a role, but it is unlikely to reverse the ischemic process if it is the only treatment offered. PMID- 8249007 TI - Clinical, radiological, and pathological aspects of cerebrovascular disease associated with antiphospholipid antibodies. The Antiphospholipid Antibodies in Stroke Study Group (APASS). PMID- 8249008 TI - Antiphospholipid antibodies. Basic mechanisms, clinical features, and animal models. PMID- 8249009 TI - Clinical, radiological, and pathological aspects of cerebrovascular disease associated with drug abuse. PMID- 8249010 TI - Black-white differences in stroke risk among young adults. AB - BACKGROUND AND PURPOSE: Little information is available regarding black-white differences in stroke incidence in young adults. METHODS: Cerebral infarction and intracerebral hemorrhage rates among adults 15 to 44 years of age were studied in Baltimore City and Baltimore County for 1988. Diagnoses were based on neurologist review of data obtained from medical records at 41 hospitals by neurological nurses. RESULTS: The cerebral infarction rates per 100,000 were 22.8 for black males, 10.3 for white males, 20.7 for black females, and 10.8 for white females. The intracerebral hemorrhage rates per 100,000 were 14.2 for black males, 4.6 for white males, 4.8 for black females, and 1.5 for white females. CONCLUSIONS: Blacks had significantly (P < .05) higher rates than whites for both cerebral infarction and intracerebral hemorrhage. Cerebral infarction rates are high in Baltimore compared with Florence, Italy, or Stockholm, Sweden. PMID- 8249011 TI - Growth factors protect neurons against excitotoxic/ischemic damage by stabilizing calcium homeostasis. AB - An aberrant elevation in intraneuronal calcium levels resulting from energy failure and excitatory amino acid receptor activation is believed to play a major role in the neuronal damage and death that occur in stroke. We have found that several growth factors can protect cultured rat hippocampal and septal neurons and human cortical neurons from excitotoxic damage caused by glucose deprivation or hypoxia. Using the calcium indicator dye fura 2 and whole-cell patch-clamp recording, we found that glucose deprivation initially results in calcium current inhibition and a reduction in intraneuronal free calcium levels without morphological signs of cell damage. After 12 to 16 hours of glucose deprivation, a large elevation in intraneuronal calcium levels occurred that involved N-methyl D-aspartate receptor activation and mediated the cell damage and death. Basic fibroblast growth factor (bFGF), nerve growth factor (NGF), and insulin-like growth factors (IGF-I and IGF-II) each prevented, in a dose-dependent manner, glucose deprivation-induced loss of calcium homeostasis and neuronal damage. The growth factors were effective to varying degrees when added up to 12 hours after the onset of glucose deprivation. NGF, bFGF, and IGFs also protected neurons against damage caused by exposure to a hypoxic environment. By stabilizing intraneuronal calcium levels within a window of concentrations conducive to neuronal survival, growth factors can protect neurons against the damaging effects of ischemia-like insults. Because ATP levels are expected to be reduced under ischemia-like conditions, we determined whether the growth factors would protect neurons against a more selective reduction in ATP levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249012 TI - Nitric oxide and nitrosothiols in cerebrovascular and neuronal regulation. AB - BACKGROUND: Currently prevailing concepts concerning the endothelium-dependent relaxant effect of acetylcholine and other endothelium-dependent agonists are that it is mediated by the generation from arginine of nitric oxide, which is then released into the extracellular space, diffuses to the vascular smooth muscle, and activates soluble guanylate cyclase by combining with the iron of the heme component of the enzyme. RESULTS AND CONCLUSIONS: Recent studies show that in the cerebral circulation these traditional concepts need to be modified in two major areas. First, the activation of soluble guanylate cyclase by nitric oxide, nitroglycerin, or nitroprusside is indirectly mediated via release of calcitonin gene-related peptide from sensory nerve fibers. This peptide then activates soluble guanylate cyclase by an unknown mechanism. Second, the endothelium derived relaxing factor from cerebral arterioles is not nitric oxide but a nitric oxide-containing compound, very likely a nitrosothiol. Nitrosothiols activate soluble guanylate cyclase in cerebral arterioles by direct action independent of calcitonin gene-related peptide. The participation of nitric oxide, nitrosothiols, or both in the regulation of basal cerebral vascular tone, in flow dependent dilation, in the vascular responses to CO2, and in response to activation of the N-methyl-D-aspartic acid receptor are considered. PMID- 8249013 TI - Distribution of atherosclerotic cerebrovascular lesions. Effects of age, race, and sex. PMID- 8249014 TI - Pathology of cerebral atherosclerosis. Influence of age, race, and gender. AB - Age, race, and gender are among the logical variables to investigate in determining the natural history of disease. In this connection, the pathological lesions of cerebral atherosclerosis have been particularly difficult to investigate. The limitations of autopsy populations, time-consuming dissections of the intracranial and extracranial arteries, and numerous technical problems associated with specimen preparation, shipment, and storage are among the impediments. Long-term repeated studies and grading systems, and the validation thereof, are also elements adding to the complexity of these studies. In the currently available publications, the most systematic studies, several of which are international in scope and from diverse medical centers, permit some tentative conclusions: (1) There is no reliable evidence of a qualitative difference in the lesions of cerebral atherosclerosis among diverse autopsy populations. (2) Quantitative differences exist in lesion severity among different age groups and races and between males and females. (3) Quantitative differences in intracranial versus extracranial atherosclerosis exist that are related to age and to race (white versus black versus Asian) and gender. (4) The role of hypertension as a factor leading to more severe and more complicated lesions is most obvious in black and Japanese autopsy populations, but its influence is not a simple one to decipher. Complicated lesions refer to stenosis, ectasias and aneurysms, thrombosis, ulceration, and calcification and hemorrhages in plaques. (5) Diet is one obvious variable differing in the populations studied so far. Cigarette smoking is probably an important factor in several populations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249015 TI - The NASCET-ACAS plaque project. North American Symptomatic Carotid Endarterectomy Trial. Asymptomatic Carotid Atherosclerosis Study. PMID- 8249016 TI - Gene polymorphism of apolipoprotein AI, the major protein of high density lipoprotein in predicting stroke risk among white and black subjects. AB - BACKGROUND AND PURPOSE: Restriction fragment length polymorphism of the apolipoprotein AI gene, which encodes the most prominent apoproteins in high density lipoprotein (HDL), were investigated using the restriction enzymes Sac I and Pst I in white and black subjects to determine the potential role of genetic variations as stroke risks as determined by carotid stenosis and an atherogenic serum profile, such as elevated total cholesterol and low density lipoprotein (LDL) levels or reduced HDL levels. METHODS: Ninety-eight subjects, including normal control subjects with no carotid stenosis and subjects with carotid stenosis, who were believed to be at stroke risk, were the study subjects and included 70 white and 28 black subjects. RESULTS: Sac I polymorphic S2 allele frequency was higher in stroke-risk groups. Significantly higher levels of serum cholesterol, triglycerides, and LDL (P < .05) and significantly lower levels of HDL (P < .05) were present in the stroke-risk group with carotid stenosis. Our study showed the following: Sac I polymorphism frequency was significantly higher in black than white subjects (chi 2 = 3.92, P < .05). Triglyceride level was significantly higher in white subjects compared with black subjects (P < .05). In white subjects, carotid artery stenosis was associated with significantly elevated total cholesterol and LDL levels (P < .01) but not with Sac I polymorphism. In black subjects, the reverse was seen with the Sac I polymorphic S2 allele associated with carotid bifurcation stenosis but did not reach statistical significance because of the small number of subjects. In addition, Sac I polymorphism did not correlate with any lipid profile. Pst I polymorphism was not associated with an abnormal atherogenic lipid profile or carotid artery stenosis abnormalities. CONCLUSIONS: Our study shows that carotid artery stenosis in white subjects is associated with increased plasma total cholesterol and LDL levels and an atherogenic profile but not with Sac I polymorphism for apoprotein AI. In black subjects, Sac I polymorphism appears to identify individuals with significant carotid stenosis, a necessary precursor to atherothrombotic brain infarction, but not those with elevated total cholesterol or LDL and/or reduced HDL levels. These results suggest that Sac I polymorphism may identify black subjects at increased risk for atherothrombotic brain infarctions. PMID- 8249017 TI - The anatomy and physiology of cortical mechanisms of cardiac control. AB - For nearly fifty years, attention has repeatedly been drawn to electrocardiographic changes accompanying acute stroke. The mechanism was unknown. These alterations, which are more frequent after hemisphere than brain stem infarction, are not solely due to concomitant myocardial ischemia. Recently, cardiac chronotropic organization has been shown within the posterior rat insular cortex. Stimulation of that site mimics both the electrocardiographic changes and the cardiac pathology accompanying acute stroke. These effects are attended by increased sympathetic activity. In addition, stimulation of the human right insula increases sympathetic cardiovascular tone, whereas parasympathetic increases are more frequent during left insula stimulation. It is suggested that right middle cerebral artery infarction disinhibits insular function, resulting in increased sympathetic cardiovascular tone and the cardiac consequences of stroke. PMID- 8249018 TI - Cerebral blood flow and metabolism in newborn infants. What can they teach us about stroke? PMID- 8249019 TI - Complement activation, neutrophils, and oxygen radicals in reperfusion injury. PMID- 8249020 TI - Positron emission tomography imaging and the therapeutic window. PMID- 8249021 TI - Applications of NMR spectroscopy to the study of experimental stroke in vivo. AB - BACKGROUND AND PURPOSE: Magnetic resonance spectroscopy and imaging enable us to investigate biochemical and pathophysiological changes associated with cerebral ischemia. The specific aims of these studies were to establish the relationships between energy metabolites and regional cerebral blood flow and to determine whether diffusion-weighted imaging is sensitive to the known thresholds for cerebral tissue energy failure and disturbance of transmembrane ionic gradients in gerbils. METHODS: Magnetic resonance spectroscopy measurements of energy metabolites in the gerbil brain were obtained as a function of cerebral blood flow (measured with the hydrogen clearance technique) before, during, and after unilateral or bilateral occlusion of the common carotid arteries. Diffusion weighted and T2-weighted images were obtained in a separate series of experiments. RESULTS: Major changes in brain energy metabolites were observed at flow values of 20 ml.100 g-1.min-1 and below. The cerebral blood flow threshold for maintenance of energy status was lowered in hypothermia, consistent with a protective effect. Diffusion-weighted imaging intensity increased at cerebral blood flow values of 15 to 20 ml.100 g-1.min-1 and below and increased gradually following the onset of severe global cerebral ischemia, but with a delay of about 2.5 minutes. CONCLUSIONS: The spectroscopic observations suggest that the flow thresholds for electrical function and edema are a direct consequence of energy failure. Comparison of the spectroscopy and imaging data suggests that diffusion weighted imaging is sensitive to disruption of tissue energy metabolism or to a consequence of this disruption. The possibilities arise of visualizing energy failure with the spatial resolution characteristic of magnetic resonance imaging and detecting compromised but recoverable tissue. PMID- 8249022 TI - Early detection of regional cerebral ischemia using high-speed MRI. AB - BACKGROUND AND PURPOSE: Diffusion-weighted magnetic resonance imaging (MRI) has been shown to be effective in detailing regions of cerebral ischemia in which water proton translations or motions have been slowed. The corresponding perfusion patterns, however, have not been correlated. Further, the hemodynamics of normal and ischemic tissues and the changes due to mild insults are also not clear. This study describes high-speed MRI techniques and observations found in the early detection of regional cerebral ischemia in the cat. METHODS: Gradient echo and spin-echo-planar MRI was used with middle cerebral artery balloon occluders to induce transient ischemia and reperfusion. RESULTS: Apparent diffusion fell within minutes after middle cerebral artery occlusion and correlated with near-total or total perfusion deficits. Reactive hyperemia, apnea, and vasodilatation appeared to be changed in ischemic and normal brain. CONCLUSIONS: Characterization of early ischemic events is dramatically improved when the motions of water (apparent diffusion), delivery of water (perfusion), and response to hemodynamic perturbations (regulatory response) are measured. All can be accomplished with high-speed MRI techniques described herein. PMID- 8249023 TI - Nuclear magnetic resonance methods in stroke. PMID- 8249024 TI - HSP70 heat shock gene regulation during ischemia. AB - The hsp70 gene is induced by denatured protein in injured cells and is an extremely sensitive and reliable marker of cells injured by ischemia, seizures, and toxins. Normal brains have little detectable hsp70 mRNA or HSP70 protein. After status epilepticus produced by systemic injections of kainic acid, however, HSP70 protein is induced in neurons but not glia in brain regions known to be injured by kainic acid. Global and focal ischemia also induce the hsp70 gene in brain. The induction of HSP70 protein in hippocampus following increasing durations of global ischemia correlates with the regional and cellular vulnerability to ischemia: CA1 neurons express HSP70 after the briefest periods of ischemia followed by CA4, CA3, dentate granule neurons, glia, and lastly, endothelial cells. Moreover, as the severity of ischemia worsens, a transcriptional and/or translational blockade of the hsp70 gene occurs in the same order so that moderate degrees of ischemia induce HSP70 in CA3 neurons and dentate granule neurons but not necrotic CA1 neurons, and severe ischemia induces HSP70 in capillary endothelial cells of hippocampus but not in any infarcted neurons or glia throughout the hippocampus. Brief periods of focal ischemia induce HSP70 primarily in neurons, suggesting that even focal ischemia can produce selective neuronal injury without infarction. In some instances, HSP70 immunoreactive astrocytes surround the HSP70 immunostained neurons. Focal ischemia that produces infarction induces HSP70 primarily in endothelial cells of cerebral blood vessels in the regions of infarction and in neurons and astrocytes on the perimeter or the penumbral area of infarction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249025 TI - Expression of immediate early gene and growth factor mRNAs in a focal cerebral ischemia model in the rat. AB - BACKGROUND AND PURPOSE: Increased release of excitatory neurotransmitters leading to activation of signal transduction has been noted after cerebral ischemia. One of the biochemical events in the signaling processes is an alteration of gene expression. A focal cerebral ischemia-reperfusion model in the rat was used to study the postischemic alteration of selected proto-oncogene and neurotrophin gene expression. METHODS: Focal ischemia in the cerebral cortex irrigated by the right middle cerebral artery was induced by temporary occlusion of the right middle cerebral artery and both common carotid arteries for 30 or 90 minutes. Expression of selected proto-oncogenes and neurotrophin genes was studied with Northern blot analysis and in situ hybridization. RESULTS: Northern blot analysis showed that an increase in the expression of selected proto-oncogenes including c fos, c-jun, jun B, zif/268, and nur 77 were noted 30 to 90 minutes into postischemic reperfusion. In situ hybridization revealed different regional distribution of c-fos mRNA signal under different experimental conditions. Nuclear run-on experiments showed that the increase in c-fos and jun B mRNA signals was due to an increase in the transcription rate. An increase in AP-1 binding activity in the ischemic cortex was also noted. Increased expression of brain-derived neurotrophic factor and nerve growth factor mRNAs were noted subsequent to the expression of immediate early genes in the ischemic cortex and adjacent areas in a biphasic pattern. CONCLUSIONS: Results are in agreement with the contention that increased c-fos and jun B expression leads to increased AP-1 binding activity, which in turn has been linked to the enhanced expression of neurotrophin genes. PMID- 8249026 TI - Inflammatory glia mediate delayed neuronal damage after ischemia in the central nervous system. AB - Reactive microglia respond within hours to central nervous system ischemic injury as exhibited by increased surface molecules, including the scavenger receptor. It is at least several days after an insult, however, before these activated mononuclear phagocytes reach a peak of secretory activity with the release of neurotoxins. This period of cytotoxin secretion is associated with a delayed neuronal loss seen in tissues neighboring sites of ischemia. Microglia suppressing drugs reduce tissue production of neurotoxic factors and improve functional outcome after ischemic injury. Immunosuppressive therapy may offer a means to reduce late neuronal damage associated with stroke. PMID- 8249027 TI - The clinical problem of intracerebral hematoma. PMID- 8249028 TI - Treatment of spontaneous intracerebral and intracerebellar hemorrhages. AB - Most primary intracerebral and intracerebellar hemorrhages are hypertensive, and the most common site is the basal ganglion. In typical basal ganglia hematoma, surgery offers no benefit, and such patients should be treated conservatively. Surgery is not indicated in pontine hematomas either. Cerebellar hematomas may block the circulation of the cerebrospinal fluid and cause an acute life threatening hydrocephalus; therefore such hematomas should be operated on. Subcortical hematomas, which are usually not associated with hypertension and may be due to tumor or vascular malformation, should as a rule be operated on. Carotid angiography is necessary for most supratentorial hematomas to exclude the presence of aneurysm or arteriovenous malformation. Secondary hematomas from ruptured arterial aneurysm should be operated on as urgently as traumatic intracranial hematomas if the patient's level of consciousness is deteriorating and if there is severe neurological deficit. Hematomas due to arteriovenous malformation must sometimes be evacuated as an emergency measure if the patient is unconscious, and the malformation should be excised if technically possible. The operation should preferably be postponed to the second week after the bleeding if the patient's level of consciousness is not deteriorating, since the malformation is more easily excised after the brain edema has subsided. Hematomas associated with anticoagulant treatment should be evacuated if the hematoma is expansive and if the patient is unconscious or somnolent but the results are not very good. Hematomas of hemophiliacs should be evacuated, and these patients need an appropriate replacement therapy. PMID- 8249029 TI - New approaches in the treatment of hypertensive intracerebral hemorrhage. AB - A comparison of the long-term outcome after surgical and nonsurgical treatment of hypertensive putaminal hemorrhage was performed in Japan over the last 15 years to determine the appropriate role of surgery. The overall results show a poor response to surgical treatment, but neurosurgeons also know that hematoma evacuation may bring about a dramatic result in some cases. In addition, experimental studies have shown that hematoma evacuation improves neuronal function at the penumbra. The discrepancy between the results of this study and the neurosurgeon's clinical impression is probably a reflection of the following. In the past, the decision to operate was determined mainly by the location of the hemorrhage as determined by computed tomography and/or magnetic resonance imaging. However, this method of deciding surgical indications is probably not correct. We are trying to change the method of determining the surgical indications from morphological to physiological criteria as follows: All patients are initially treated with hyperbaric oxygen, and those who show improvement of their symptoms are clearly indicated for surgery. If the somatosensory evoked potential or auditory brain stem response shows an improvement after administration of mannitol or glycerol, this is also an indication that surgery should be performed. PMID- 8249030 TI - Stressful life events, psychopathology, and adolescent suicide: a case control study. AB - The relationship between stressful life events and adolescent suicide was assessed in 67 adolescent suicide victims and 67 matched community controls. In the year before death, suicide completers were more likely to have experienced: (1) interpersonal conflict with parents and with boy/girlfriends, (2) disruption of a romantic attachment, (3) legal or disciplinary problems. Legal or disciplinary problems were more commonly associated with suicide in conduct and substance abuse disordered youth. Interpersonal loss was more commonly associated with suicide in substance abuse as well. Even after controlling for psychopathology, legal and disciplinary problems in the past year were associated with an increased risk of suicide. PMID- 8249031 TI - Use of the Suicide Probability Scale (SPS) with adolescents. AB - This study examined the utility of the Suicide Probability Scale (SPS) by comparing results obtained with a sample of 217 normal adolescents (M = 16.2 years) to the findings obtained in Cull and Gill's (1988) standardization effort. The present sample scored significantly higher than Cull and Gill's normative sample in SPS item, subscale, and total scores. In addition, the SPS generally failed to discriminate between the present, adolescent sample and Cull and Gill's inpatient psychiatric sample. Moreover, item-subscale correlations obtained for the present sample often differed from those reported by Cull and Gill, and factor analysis of SPS item scores failed to replicate Cull and Gill's four factor solution. Findings suggest the need for caution when using the SPS to assess adolescent suicide potential and underscore the need for additional research regarding the instrument's efficacy in distinguishing between normal and suicidal adolescents. PMID- 8249032 TI - Sudden death and bereavement outcomes: the impact of resources on grief symptomatology and detachment. AB - The study of sudden bereavement focuses almost exclusively on its impact on psychological well-being. This research broadens our understanding of bereavement outcomes by exploring the effect of grief on family detachment following sudden bereavement. Moreover, this study examines the relative contributions of resources in alleviating grief symptoms and reducing detachment. Data are collected from medical examiner records and mail-back surveys from family members of victims of suicide and accidental deaths in a large metropolitan area of the United States. A covariance structural equation model allowing for correlated measurement error is estimated using LISREL VII. The results indicate that the most grief-stricken survivors are more detached from family than those who are least grief-stricken. Examination of the direct and indirect influences of resources on bereavement outcomes reveals that self-esteem is the strongest predictor of grief and that it also plays a decisive role in mediating the influences of religious participation and exercise on grief. Expressive support exerts direct effects on both grief and detachment. This research concludes by directing future research to examine more fully the impact of sudden bereavement on marital and family relationships. PMID- 8249033 TI - The relationship between coping strategies and perceived stress in telephone intervention volunteers at a suicide prevention center. AB - This study examines stress perceived by telephone intervention volunteers at a suicide prevention center before their shift, during the most high-urgency call, and after the shift. Eighty of the 82 active volunteers completed questionnaires concerning stress, coping strategies, motivations for volunteer work, and experiences with suicide. Stepwise multiple regression analysis indicated that only one variable, the amount of experience in telephone intervention with suicidal persons, predicted stress level before the shift; volunteers with more experience tended to be less stressed. Stress during the most urgent call was related first to the level of urgency of the call, then to the total length of all calls received, followed by the coping mechanisms of magical thinking, detachment, and feeling personally responsible. Stress after the shift was related first to the total amount of time spent on calls, then to the number of other persons present during the shift. Stress after the shift was also negatively related to amount of education and having realistic expectations about interventions. Magical thinking was positively related to stress, and the mechanism of positive thinking was negatively related. A high proportion of volunteers had attempted suicide, had previous thoughts about suicide, and had known persons who attempted or died by suicide. These findings are discussed as to their implications for the selection and training of volunteers in suicide prevention. PMID- 8249034 TI - Potential for suicide and aggression in delinquents at Juvenile Court in a southern city. AB - Because juvenile delinquents rarely communicate their wishes for suicide or homicide appropriately to busy caretakers, some means to identify those juveniles at greatest risk of acting out was sought. Psychological examiners in a Southern city individually questioned 263 Juvenile Court offenders about whether they wished to be dead, to kill themselves, or to kill others; and also about their existential experiences, thoughts and feelings. Taking the data associated with each wish, we used stepwise multiple logistic regression to pinpoint experiences, such as substance abuse, that were associated with a high likelihood of verbalizing a wish for destructive behavior. These results included sexual abuse as a risk factor and the presence of a mother or grandmother as a protective factor, and led to recommending education of caretakers and forming a screening test. PMID- 8249035 TI - Inpatient standards of care and the suicidal patient. Part I: General clinical formulations and legal considerations. AB - The most common legal action involving psychiatric care is the failure to reasonably protect patients from harming themselves. In this regard it is critical to understand that courts have tended to impose much stricter standards on inpatient than on outpatient care; that at the present time, most malpractice actions involve clinical activities related to inpatient care (negligent admission, treatment, supervision, discharge, etc.). This article reviews the current climate in the legal and clinical formulation of standards of care for hospitalized adult suicidal patients. It suggests general guidelines for effective assessment, management, and treatment procedures that balance the need for high-quality care by a reasonable and prudent practitioner with the requirements of court-determined and statutory standards. The authors specifically discuss court cases that show common failure situations in inpatient care, discharge planning, and follow-up (e.g., problems in pharmacotherapy, the decision to hospitalize, the assessment of imminence and lethality, etc.). The paper also emphasizes the crucial element of clinical judgment in developing any inpatient standard of care. PMID- 8249036 TI - The suicide of Anne Sexton. AB - Anne Sexton, a distinguished poet, killed herself in 1974. Sexton's poems, many of which deal with suicide, read together with a richly researched recent biography--enhanced by the biographer's access to tapes of Sexton's sessions with her psychiatrist and by a foreword by that psychiatrist--permit an unusual opportunity to understand the interrelationship of her illness, her treatment, and the meaning of suicide in her life. PMID- 8249037 TI - The effectiveness of suicide prevention centers. AB - The presence of suicide prevention centers in a state in 1970 was negatively associated with changes in the suicide rate from 1970 to 1980 in the USA, indicating a preventive effect from suicide prevention centers. This effect, though weak, was consistently found for most demographic subgroups of the population and when a strong social correlate of suicide rates (divorce rates) was taken into account by means of multiple regression analysis. PMID- 8249038 TI - Suicide in the workplace. PMID- 8249039 TI - Case of Kim. PMID- 8249040 TI - Plasma concentrations of dothiepin and its metabolites are not correlated with clinical efficacy in major depressive illness. AB - Plasma concentration-antidepressant response relationships for dothiepin, nordothiepin, dothiepin-S-oxide, and nordothiepin-S-oxide were investigated in 50 patients (33 women and 17 men), who had had a major depressive episode. Depression and anxiety were assessed at the start of therapy and after 2 and 4 weeks by measurement of a Hamilton rating score for depression (HRSD), a Beck depression inventory (BECK), visual analog scores for depression (VASDEP) and anxiety (VASANX), and a physician's global (GLOBAL) score. There were significant (p < 0.001) decreases in both mean depression (32-69%) and mean anxiety (30-44%) scores at weeks 2 and 4, but there were no robust linear or polynomial correlations between percent decrease in depression or anxiety scores and plasma concentrations of dothiepin or its metabolites at week 4. It is suggested that measurement of the nordothiepin/dothiepin ratio may assist in the assessment of compliance. PMID- 8249041 TI - Nicotine, cotinine, and trans-3-hydroxycotinine levels in seminal plasma of smokers: effects on sperm parameters. AB - Sperm samples from 44 cigarette smokers and 50 nonsmokers attending an infertility clinic were examined by high-performance liquid chromatography (HPLC) assay and HPLC-mass spectrometry for the presence of nicotine (NIC), cotinine (COT), and trans-3'-hydroxycotinine (THOC) in seminal plasma. Smokers were found to have levels of COT and THOC in seminal plasma that were similar to those found in serum. The level of NIC was significantly increased in seminal plasma compared to serum. Total motility of spermatozoa was significantly and negatively correlated to COT and THOC levels in seminal plasma. Forward motility of spermatozoa was correlated only with cotinine semen levels. On the basis of these results, we suggest that the presence of tobacco smoke constituents in seminal plasma could provide a warning of the adverse effects of cigarette smoke on the physiology of reproduction. PMID- 8249042 TI - Morphine pharmacokinetics and effects on salivation and continuous reaction times in healthy volunteers. AB - Ten healthy volunteers were given an i.v. infusion of 10 mg morphine HCl, an oral solution of 20 mg morphine HCl, or a new controlled release tablet of 30 mg morphine sulphate on three separate occasions in a complete crossover design. Venous blood samples were collected serially for 14-24 h and analyzed for morphine using high-performance liquid chromatography (HPLC). Continuous reaction times (CRTs) and salivation were measured repeatedly in all subjects. Oxygen saturation remained normal throughout the procedure. Five subjects experienced nausea on at least one occasion. Pharmacokinetic parameters, calculated using a two-compartment model, were in accordance with previous results for i.v. and oral administration of morphine solutions. The absolute bioavailability of morphine in the oral solution was 21.6% (15.4-27.7%; 95% CI) and in the controlled release tablet, 17.1% (12.6-21.6%; CI). Secondary peaks in the plasma concentration curves strongly indicated an enterohepatic circulation (EHC) of morphine. Alternative pharmacokinetic calculations, including EHC, were performed and used in a pharmacokinetic-pharmacodynamic model, in which the studied effects were well correlated to the concentrations of morphine. PMID- 8249043 TI - Pharmacokinetic interaction between high-dose methotrexate and amoxycillin. AB - A case report of toxicity following concurrent administration of high-dose methotrexate and amoxycillin is presented. A 16-year-old male patient was administered 10 high-dose methotrexate cycles for treatment of a fully malignant osteogenic sarcoma. Methotrexate was administered at a dosage of 8 g/m2 and infused intravenously over a 6-h period. The patient received pre- and posttreatment hydration and sodium bicarbonate for alkalinization of urine. Calcium folinate rescue was performed when appropriate. During the 10th cycle, coadministration of amoxycillin (1 g/6 h, p.o.) resulted in prolonged and marked enhancement of methotrexate serum levels. Pharmacokinetic parameters obtained in cycle 10 indicate significant differences for total plasma clearance, mean residence time, and distribution half-life when compared to those in cycles 1-9. Amoxycillin decreased the renal clearance of methotrexate, probably by competition at the common tubular secretion system and by secondary methotrexate induced renal impairment. The patient experienced acute and subacute toxicity with renal failure, myelosuppression, mucositis, nausea, vomiting, fever, and dermatologic abnormalities. Patients receiving amoxycillin during methotrexate therapy should be closely monitored to avoid severe toxicity. PMID- 8249044 TI - Individualizing drug dosage regimens: roles of population pharmacokinetic and dynamic models, Bayesian fitting, and adaptive control. AB - The role of population pharmacokinetic modeling is to store experience with drug behavior. The behavior of the model is then correlated with the clinical behavior of the patients studied, permitting selection of a specific serum level therapeutic goal that is based on each individual patient's need for the drug and on the risk of adverse reactions, both of which must be considered. A dosage regimen is then computed to achieve that goal with maximum precision. The patient should not run a greater risk of toxicity than is justified, and should obtain the maximum possible benefit within the acceptable risk. The regimen is given and the patient monitored. PMID- 8249045 TI - Development and validation of a limited sampling strategy for 5-fluorouracil given by bolus intravenous administration. AB - A wide range of interindividual variability of 5-fluorouracil (5-FU) pharmacokinetics exists after bolus administration. The degree to which this variability in 5-FU exposure impacts upon the response and toxicity of the drug has not been determined. The area under the concentration time curve (AUC) is a commonly used indicator of exposure, but normally requires the collection of 8-10 timed blood samples after i.v. bolus administration. This presents difficulties if large-scale population samplings are required. This study involved the development and testing of a strategy to estimate AUC from a limited number of blood samples in patients with gastrointestinal and breast cancer. The optimal single time point for AUC estimation was 0.17 h (r2 = 0.954). Addition of the 0.75 h time point significantly improved predictability (r2 = 0.983). Addition of a third or fourth time point did not provide further benefit. These models were then tested separately in a group of women who received a higher dose of 5-FU. The two data points model performed significantly better than the single time point model (r2 = 0.70 and 0.85, respectively). The AUC of standard dose 5-FU after bolus administration can be reliably estimated from two timed samples taken 10 and 45 min after injection. PMID- 8249046 TI - Single-dose pharmacokinetics of the new immunosuppressant RS-61443 in the rabbit. AB - The pharmacokinetics of mycophenolic acid (MPA) were investigated in six New Zealand white rabbits following (i.v.) and (s.c.) administration of RS-61443. Plasma concentrations of MPA were analyzed by high-performance liquid chromatography (HPLC). Model-dependent and -independent parameters were calculated. Mycophenolic acid is rapidly cleared with a terminal half-life of approximately 1.0 h. The volume of distribution subsequent to i.v. administration indicates minimal distribution into tissues. The mean residence time and volume of distribution were greater for the s.c. as compared to the i.v. route of administration. The insignificant difference in the area under the curve between the two dosage routes infers that the bioavailability of RS-61443-derived MPA by the former route of administration is comparable to the latter. Trough concentrations at the doses administered are below the detection limit of the HPLC procedure. The implications of these findings in relation to the therapeutic monitoring of the drug are discussed. PMID- 8249047 TI - Evaluation of the effect of fluoxetine on the formation of carbamazepine epoxide. AB - Fluoxetine has been reported to increase carbamazepine (CBZ) plasma concentrations and cause adverse effects. CBZ-10, 11 epoxide (CBZE), the major metabolite of CBZ, contributes to the clinical effect and toxicity of CBZ. The objective of the present study was to investigate the effect of fluoxetine and its major metabolite, norfluoxetine, on CBZE formation in isolated perfused rat liver, in vitro human liver (n = 5) microsomes, and patients (n = 14), after either CBZ monotherapy or polytherapy with fluoxetine. In isolated perfused rat liver, there was no effect of fluoxetine (n = 8) or norfluoxetine (n = 6) on the formation clearance of CBZE (12.8 +/- 5.3 and 11.7 +/- 3.8 ml/min, respectively) or the intrinsic metabolic clearance of CBZ (6.6 +/- 2.7 and 6.3 +/- 1.8 ml/min, respectively). Studies on human liver microsomes confirmed that neither fluoxetine or norfluoxetine inhibited formation of CBZE until concentrations were > 20 times those found clinically. In support of this, there was no difference in the ratio of CBZE to CBZ plasma concentrations in patients also receiving fluoxetine when compared to patients on CBZ monotherapy; however, there was a trend toward a decrease in the apparent plasma clearance of CBZ. In conclusion, increased plasma concentrations of CBZ found when fluoxetine is added are not due to decreased formation of CBZE. Clinically, if fluoxetine causes an increase in CBZ levels, CBZE plasma concentrations will increase proportionately and contribute to the toxicity. PMID- 8249048 TI - Experiences with the enzyme-multiplied immunoassay cyclosporine specific assay in a therapeutic drug monitoring laboratory. AB - Cyclosporin-A (CsA) monitoring is well established in the management of most organ transplant patients. The present communication reviews the performance of the recently introduced specific enzyme-multiplied immunoassay (EMIT) CsA method during the first 7 months of its operation and compares costs of providing this service with those of the specific 125I radioimmunoassay (RIA) method previously employed in this clinical laboratory. Results suggest that the EMIT method performed well, giving long calibration curve stability (up to 12 weeks), and only 4.4% of the 31 kits through this period were consumed in assaying calibration standards compared with 20.8% with RIA. However, more quality control assays were performed, with the net result that only a slight improvement in the percentage of kit consumed in patient assays was noted (74.0% compared with 70.3%) with the EMIT method. This method appears to have been well accepted clinically as the CsA assay request rate over this period increased by 23% and, since it is both specific and rapid, is, therefore, recommended as the best CsA method currently available. PMID- 8249049 TI - Influence of serum separator tubes on total and free phenytoin concentrations and dosages. AB - The influence of serum separator tubes (SSTs) on total and free phenytoin concentrations and phenytoin dosages was evaluated in patients treated with phenytoin. Thirty blood samples were obtained from 18 patients. Equal volumes of blood were placed into SSTs and plain tubes. Samples were centrifuged, and serum analyzed for total and free phenytoin by fluorescence polarization immunoassay (FPIA) at 2, 24, and 48 h. Total phenytoin concentrations collected in SSTs were significantly lower than those collected in plain tubes at 2 (12.3 vs 12.8 mg/L, p < 0.01), 24 (11.4 vs 12.9 mg/L, p < 0.01), and 48 h (10.9 vs 12.9 mg/L, p < 0.01). These differences resulted in significantly higher calculated maximum rates of metabolism (Vmax) and daily phenytoin dosages (R0) for SSTs at 24 (Vmax: 10.4 vs 10.1 mg/kg/day; R0: 8.2 vs 8.0 mg/kg/day, p < 0.01) and 48 h (Vmax: 10.8 vs 10.3 mg/kg/day; R0: 8.5 vs 8.1 mg/kg/day, p < 0.01). Free phenytoin concentrations from SSTs were significantly lower at 48 h (1.56 vs 1.61 mg/L, p < 0.05). However, there were no significant differences in calculated dosages. Observed statistical differences in total phenytoin concentrations can be clinically important for making dosage adjustments, especially in patients undergoing nonlinear elimination. Thus, SSTs should not be used to collect blood for total serum phenytoin determination. PMID- 8249050 TI - Saliva nicotine as an index of plasma levels in nicotine skin patch users. AB - This study examined whether salivary nicotine concentrations would provide a useful index of plasma concentrations in studies of the effects of transdermal nicotine administration. Twenty-four subject smokers abstained from smoking for 12 h prior to admission to a clinical research unit ward and for 36 h while remaining confined to the ward. Three doses of nicotine skin patches were applied to different groups of subjects, and blood and saliva samples were collected at several time points. Saliva flow was stimulated by three different methods: (a) sucking on a lemon candy, (b) dissolving a sugar cube in the mouth, and (c) chewing on parafilm. Nicotine and cotinine concentrations were measured in both saliva and blood using gas chromatography. There was a high correlation between blood and saliva values of nicotine (r = 0.82). Overall, saliva nicotine concentrations were approximately 8.1 times higher than those of plasma. Saliva and blood cotinine concentrations were also highly correlated (r = 0.94), replicating results of previous studies. Results suggest that saliva nicotine may be a useful marker of nicotine intake during nicotine skin patch treatment. Saliva collection may be advantageous under conditions in which blood collection is impractical and may provide greater sensitivity because of the high concentration of nicotine in saliva relative to that in blood. PMID- 8249051 TI - Immunoassay reagents for psychoactive drugs. Part 3. Removal of phenothiazine interferences in the quantification of tricyclic antidepressants. AB - Phenothiazines and their metabolites are known to interfere in the quantification of tricyclic antidepressants (TCAs). A method for selective chemical modification of phenothiazines by chloramine-T in the presence of TCAs is described. This method allows for accurate quantification of the TCA analyte in a serum sample without interference from the modified phenothiazine. PMID- 8249052 TI - Determination of the serum protein binding of oxycodone and morphine using ultrafiltration. AB - Protein binding of oxycodone and morphine in human serum was determined in vitro using ultrafiltration. Binding studies were also performed using both purified human serum albumin and human alpha 1-acid glycoprotein (AAG). Albumin was found to be the major binding protein for both oxycodone and morphine. The serum protein binding of both oxycodone and morphine was independent of drug concentration in the therapeutic range (5-100 ng/ml), but was dependent on protein concentration. In addition, bound fractions of oxycodone and morphine increased with increasing concentrations of both albumin and AAG. At physiological pH and temperature, the mean (+/- SD) serum protein binding of oxycodone was 45.1% (+/- 0.4%) and that of morphine was 35.3% (+/- 0.2%) A decrease in temperature from 37 to 23 degrees C significantly increased the serum protein binding of oxycodone and morphine by 8-9% (p < 0.0001) and 7-10% (p < 0.0001), respectively, indicating the importance of maintaining the temperature at 37 degrees C during protein binding experiments. A reduction in pH from 7.75 8.85 to 7.4 significantly reduced serum protein binding of both oxycodone and morphine by 4-5% (p < 0.0001) and 4-7% (p < 0.0001), respectively. Serum samples, to which known concentrations of oxycodone had been added and which were stored at -20 degrees C, showing a gradual but significant decline (p < 0.0001) in serum protein binding of oxycodone from approximately 45 to 39% during the 4-week storage period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249053 TI - Enzyme immunoassays for a new angiotensin-converting enzyme inhibitor, zabicipril, and its active metabolite in human plasma: application to pharmacokinetic studies. AB - Zabicipril (S 9650) is a new angiotensin-converting enzyme inhibitor whose hydrolysis in vivo produces the pharmacologically active metabolite zabiciprilat (S 10211). Two competitive enzyme immunoassays specific for either zabicipril or zabiciprilat have been developed using acetylcholinesterase (E.C. 3.1.1.7) as label. Antibodies were raised in rabbits after immunization with lysil derivatives of zabicipril or zabiciprilat coupled with bovine serum albumin. Assays were performed in 96-well microtiter plates coated with a monoclonal antibody raised against rabbit immunoglobulin G, thus ensuring rapid separation of free and bound fractions of the tracer. The analysis does not require any extraction step. In the case of the assay of zabiciprilat, interference generated by endogenous angiotensin-converting enzyme (ACE) was eliminated by the addition of perindoprilat, another ACE inhibitor. Perindoprilat was not recognized by the antibodies (cross-reactivity < 0.01%) and did not affect assay efficiency. The specificity of the assays was checked by high-performance liquid chromatography of human plasma samples obtained after oral administration of 2 mg of zabicipril. No metabolites or endogenous substances were detected. The mean reproducibility was 15% for the assay of zabicipril and 19% for the assay of zabiciprilat. The quantification limits were 1.2 ng/ml for the zabicipril assay and 0.8 ng/ml for the zabiciprilat assay. These assays are therefore suitable for pharmacokinetic studies and drug monitoring in clinical studies. PMID- 8249054 TI - A sensitive and simple high-performance liquid chromatographic method for the determination of doxorubicin and its metabolites in plasma. AB - A sensitive high-performance liquid chromatographic (HPLC) method for the measurement of doxorubicin and its metabolites in plasma is described. After precipitation with zinc sulphate and methanol, samples are resolved by isocratic elution from a C18 reverse phase support within 20 min and quantified by endogenous fluorescence. Recoveries over a concentration range from 5 to 1,000 nM of doxorubicin, doxorubicinol, doxorubicinone, doxorubicinolone, and 7 deoxydoxorubicinolone were 80-110%, while recovery for 7-deoxydoxorubicinone was approximately 60%. At concentrations of 5 nM, within-run and between-day coefficients of variation for each compound were < 8 and < 16%, respectively. Limits of detection for the compounds were 1-2 nM and standard curves were linear up to at least 1,000 nM. The drug and its metabolites are stable in deproteinized plasma samples at room temperature and in the dark for at least 24 h. The method requires few manipulations and is readily adaptable to automated analysis of large series of samples. PMID- 8249055 TI - Comparison of theophylline blood levels measured by the standard TDx assay and a new patient-side immunoassay cartridge system. AB - Plasma steady-state concentrations of theophylline were measured by the standard TDx assay and a novel patient-side immunoassay system with a single-use cartridge (Biotrack 516) in 100 samples from 93 patients. Intra- and interassay coefficients of variation for the new cartridge technique averaged 4.3 and 9.8%, respectively. In the concentration range of 2 to 40 micrograms/ml, there was a linear correlation (r = 0.96, p < 0.0001) between both assays [slope of 0.86; y intercept (TDx values) = -0.14 micrograms/ml], indicating that the two methods can be used interchangeably. Since the completely automated cartridge system is very simple and extremely rapid, this technique offers some advantages for routine clinical use, especially under ambulatory conditions. PMID- 8249056 TI - Immunization in the developing world: strategic challenges. PMID- 8249057 TI - Royal Society of Tropical Medicine and Hygiene meeting at Manson House, London, 10 December 1992. George Carmichael Low FRCP: twelfth president of the Society and underrated pioneer of tropical medicine. AB - George Carmichael Low, like so many early pioneers of Tropical Medicine, had his origin(s) in Scotland. Following a distinguished undergraduate (and early postgraduate) career, he joined Dr Patrick Manson at the newly established London School of Tropical Medicine in 1899. His first major contribution to the specialty (in 1900) was to demonstrate filariae in the proboscis sheath of mosquitoes which had been infected with Filaria bancrofti in Australia, using a technique recently learned in Heidelberg and Vienna. Shortly afterwards, he led an expedition to the Roman Campagna; this established beyond doubt mosquito transmission of Plasmodium vivax infection to Homo sapiens. In 1901-1902, Low undertook a demanding tour of the Caribbean where he made important contributions to the understanding of the filariases, and assisted in malaria eradication. In 1902 he led a small team (the Royal Society's first sleeping sickness expedition) to investigate the 'negro lethargy' which had emerged in epidemic proportions on the northern shores of Lake Victoria in East Africa. This expedition narrowly failed to establish the aetiological agent (Trypanosoma sp.) of this disease. Following his return to London, Low became superintendent of the Albert Dock Hospital and from then onwards devoted most of his career to the London School of Tropical Medicine and the Hospital for Tropical Diseases (where he became senior physician). He wrote extensively, in addition to his clinical, teaching and administrative commitments. Perhaps Low's major contribution, however, was in establishing the Society (later Royal) of Tropical Medicine and Hygiene in 1907, with Mr (later Sir) James Cantlie.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249058 TI - Periodicity and space-time clustering of severe childhood malaria on the coast of Kenya. AB - Traditionally malaria epidemiology has focused on factors such as parasite rates and vector dynamics without specific reference to disease. There are limited comprehensive data on malaria as a life-threatening event in African children. We have identified, through hospital surveillance, 581 episodes of severe malaria in residents of a defined area on the Kenya coast over a period of 3 years. This represents an absolute minimum risk of developing severe malaria by the fifth birthday of 1 in 15. The presentation of severe malaria showed marked seasonality, but the timing and magnitude of these fluctuations varied considerably between years. A satellite navigational system was used to define the exact location of the home of each severe malaria case. Space-time clustering of severe malaria was evident in this community. Seasonal peaks in incidence of severe malaria may comprise discrete mini-epidemics. In contrast, parasite rates in the community varied little during the course of the surveillance. The monitoring of disease, as opposed to parasitization, in children may result in more effective targeting of intervention resources. PMID- 8249059 TI - Anophelines in the state of Acre, Brazil, infected with Plasmodium falciparum, P. vivax, the variant P. vivax VK247 and P. malariae. AB - Anophelines collected indoors and in the peri-domiciliary area in 3 localities in the Amazon region, state of Acre, Brazil, from August 1990 to January 1991 were examined by enzyme-linked immunosorbent assay (ELISA) using specific monoclonal antibodies directed against the repeats of the circumsporozoite proteins of Plasmodium falciparum, P. vivax, P. vivax V247, and P. malariae. Of the 3056 specimens collected, 2610 were Anopheles oswaldoi, 362 A. deaneorum, 60 A. triannulatus and 24 were A. darlingi. The infection rates of A. oswaldoi were 3.41% for P. falciparum, 2.26% for P. vivax, 1.22 for P. vivax VK247, and 0.42% for P. malariae. For A. deaneorum, the infection rates were 2.76% for P. falciparum, 0.55% for P. vivax, and 0.82% for P. vivax VK247. All samples of the other 2 species collected (A. triannulatus and A. darlingi) were negative in the ELISA. There were certain differences in the anopheline distribution and infection rates between these localities, and in one only A. oswaldoi was found to be infected. These results strongly point to A. oswaldoi as the main malaria vector in the region. No difference was found between the potential vectors of P. vivax and P. vivax VK247. The significance of these findings for malaria control is discussed. PMID- 8249060 TI - 'Baggage malaria' in Italy: cryptic malaria explained? PMID- 8249061 TI - Kala-azar in western Upper Nile province in the southern Sudan and its spread to a nomadic tribe from the north. AB - Since the start in 1988 of the present epidemic of kala-azar (visceral leishmaniasis) in western Upper Nile state in southern Sudan, the epidemiology of the disease in all parts of the Sudan where kala-azar has been reported was reassessed by the Leishmaniasis Research Group in Khartoum. In this paper, the spread of the epidemic is described among a nomadic tribe originating from southern Kordofan state, who migrate every year with their cattle to the Bentiu area in western Upper Nile state where the epidemic is still raging. 200 cases from this tribe were seen in Khartoum; another 56 cases were found during a field trip to the area. In addition, the Bentiu area was visited, where 301 cases were under treatment and another 52 of 1120 individuals screened were confirmed parasitologically. 20 cases of post-kala-azar dermal leishmaniasis were found. Parasites isolated from the nomadic tribe were of the same zymodeme as parasites isolated previously from the Nuer in western Upper Nile. The epidemiological findings in each state are discussed in relation to the tribes that were affected and the ecology of the area. PMID- 8249062 TI - Phlebotomus (Larroussius) perfiliewi naturally infected with dermotropic Leishmania infantum at Tenes, Algeria. PMID- 8249063 TI - An evaluation of factors affecting Helicobacter pylori prevalence in Tibetans exiled in India. AB - Helicobacter pylori infection is found world-wide although the epidemiology of infection has not been well defined in many geographical areas. The aims of this study were to determine the prevalence of H. pylori infection and chronic gastritis and the demographic correlates of infection in a single racial group in southern India. The sample population was selected randomly from the male population register of a Tibetan refugee settlement. Demographic data and results of endoscopy with antral mucosal biopsy were evaluated in 197 subjects (median age 28, range 21-81 years). H. pylori was present in 77.2% of subjects by histology and/or urease test. Chronic gastritis and H. pylori were closely related and there was an association between the number of bacteria present and the severity of the gastritis (P < 0.04). Infection with H. pylori was inversely associated with socio-economic factors, specifically educational level (P < 0.02) and occupation (P < 0.02). Unlike other studies, the prevalence of H. pylori was not found to rise with age, being lower in those older than 40 years (P < 0.005). This difference was still apparent when adjusted for socio-economic status. The major demographic difference between younger and older subjects of low socio economic status was the greater proportion of early life spent outside India (and in Tibet) by older subjects. Among younger subjects, residence in India for 20 years or more was associated with a greater risk of H. pylori infection (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249064 TI - Foci of Schistosoma mansoni in Assiut province in middle Egypt. AB - Following detection of Schistosoma mansoni in a 12 years old boy who had both schistosomal polyposis and periportal fibrosis with hepatosplenomegaly, epidemiological studies to confirm local transmission were performed in his and 30 other villages in Assiut Governorate, Egypt. The index case's brother and 6 of 380 inhabitants of his village who provided stool specimens were infected with S. mansoni and a farmer with dysentery and hepatosplenomegaly had a positive rectal snip biopsy. All had hepatic abnormalities on ultrasound examination. Two of 221 schoolchildren in another village had mixed infections with S. mansoni and S. haematobium; 17 others had only S. haematobium. None of 419 inhabitants living near the infected boys had S. mansoni infection. Snails from canals and drains near both villages were netted, identified, counted and checked for infection: in the second village one of 1039 Bulinus truncatus was infected with Schistosoma sp. but none of 859 Biomphalaria alexandrina was infected. Schistosomiasis mansoni is being focally transmitted in 2 villages in Assiut Governorate and appears to be spreading from Lower to Middle and Upper Egypt. We believe B. alexandrina, which has been present in some of the waterways for at least 15 years, were infected recently by local inhabitants returning from Iraq or by cattle traders or military recruits from the Delta. PMID- 8249065 TI - Modelling the efficacy of prophylactic and curative therapies for preventing the spread of tuberculosis in Africa. AB - Concerns have been raised about whether the interaction between tuberculosis and human immunodeficiency virus (HIV) may lead worldwide to a recrudescent tuberculosis pandemic. These concerns are particularly grave in Africa which has a high prevalence of both tuberculosis and HIV. This study used a computer simulation model to examine the effect of tuberculosis-HIV interactions on tuberculosis prevalence and mortality in Africa. The model then assessed the impact of expanding treatment and chemoprophylaxis programmes on tuberculosis prevalence and mortality over the next decade. In communities where 20% of the population is infected with HIV and 25% receive treatment for tuberculosis, deaths from tuberculosis would be 100% higher than in communities where none of the population is HIV-infected. In a population the size of Uganda's, during one decade there would be approximately an additional 530,000 deaths from tuberculosis. When 50% of patients with active tuberculosis receive treatment, one death will be averted for every 2.5 people who receive treatment. The prevalence of active tuberculosis could be cut by over 90% in a decade by providing effective chemoprophylaxis to 30% of individuals with inactive TB. In conclusion, TB is only one example of a preventable and treatable infectious disease which can be spread through casual contact and which, because of its higher prevalence among the HIV positive population, may lead to a preventable increase in incidence of infection among the general population. PMID- 8249066 TI - Subclinical infection with Mycobacterium leprae--a problem for leprosy control strategies. AB - Tests for the serodiagnosis of leprosy, including those based on phenolic glycolipid-I (PGL-I), have shown poor specificity for leprosy in studies of endemic communities, despite initially promising results in studies of selected patients. During a 5 years follow-up study of a hyperendemic community in Papua New Guinea, a marked reduction in numbers of seropositive children and an increase in age of those seropositive followed introduction of multi-drug therapy. This was accompanied by a reduced case detection rate and a shift to paucibacillary disease in new cases, consistent with a reduction in transmission. Only a minority of persistently seropositive persons developed leprosy. These observations suggest that subclinical infection with Mycobacterium leprae is common in endemic communities and that PGL-I seropositivity is a marker of subclinical infection, with poor specificity for overt disease. Detection of subclinical infection may confound control strategies which use screening tests to identify asymptomatic highly infectious cases for earlier therapy. PMID- 8249067 TI - Melioidosis in Papua New Guinea: is it less common than in tropical Australia? PMID- 8249068 TI - Infant disease patterns in northern Cameroon. AB - The epidemiology of common diseases of infants seen in district health service clinics in a culturally heterogeneous population in northern Cameroon was studied. Significantly higher incidences were found for respiratory tract infection in infants of non-Moslem households, and for diarrhoea in infants of Moslem households. A subsequent interview survey of parents of children aged 0-5 years revealed a higher prevalence of smokers and millet stalk kitchens in the compounds of non-Moslems. Other possible risk factors were explored. The findings indicated that in this area the influence of culture on disease in infants is strong, and epidemiological differences among cultural groups should be considered in the design of disease control programmes in the community. PMID- 8249069 TI - A population-based health survey in Kon Dieng District, Cambodia. PMID- 8249070 TI - Presumptive diagnosis of malaria in infants in an endemic area. PMID- 8249071 TI - Field application of an ELISA using redefined Leishmania antigens for the detection of visceral leishmaniasis. AB - Two soluble antigens from Leishmania donovani of 116 kDa and 70 kDa molecular mass, and a soluble mixture of crude antigens, were used in an enzyme-linked immunosorbent assay (ELISA) for the detection of visceral leishmaniasis (VL) in the field, and compared with the direct agglutination test (DAT). The tests were carried out on 8 VL patients, 34 normal individuals from an area endemic for the disease, and 68 former visceral leishmaniasis patients 1-5 years after treatment. The 70 kDa ELISA and the DAT had a sensitivity and specificity of 100% (95% confidence interval 63-100%), while the 116 kDa ELISA and the soluble crude antigen ELISA were 37.5% (9-76%) and 50% (16-84%) sensitive, respectively. When using ELISA (116 kDa or 70 kDa), 68-69% of sera tested 1-2 years, and 92-94% of sera tested 5 years, after treatment were negative. In contrast, when DAT or ELISA with crude antigen were used, the negativity rate was 31% 1-2 years, and 53% 5 years, after treatment. DAT was therefore not an accurate test for diagnosis in the field. The use of the 70 kDa antigen in ELISA was an accurate alternative to DAT in the detection of VL. PMID- 8249072 TI - Leishmanin and tuberculin sensitivity in leishmaniasis in the Sudan, with special reference to kala-azar. AB - The application of leishmanin and tuberculin skin tests was studied in patients with leishmaniasis in the Sudan. 35 cases of active kala-azar and 3 relapse cases were leishmanin negative. 81% of patients treated for kala-azar showed a positive reaction after 6 months. 17 of 29 patients with post kala-azar dermal leishmaniasis (PKDL) were leishmanin positive. 2 of 16 patients with kala-azar tested with tuberculin were positive; one was diagnosed as tuberculosis. In 7 initially tuberculin negative patients, the tuberculin test became positive after treatment. A new Leishmania major skin test antigen (Pasteur Institute, Iran) was more reactive than other antigens in patients with cutaneous leishmaniasis, mucocutaneous leishmaniasis and PKDL, but not in treated kala-azar cases. In a field study in an area of endemic kala-azar, the new L. major antigen proved more reactive both in individuals previously exposed to L. major (causing cutaneous leishmaniasis) and in those with past exposure to L. donovani. The literature concerning skin testing with leishmanin and tuberculin in kala-azar is reviewed. PMID- 8249073 TI - Diagnosis of Giardia duodenalis infection in Bangladeshi infants: faecal antigen capture ELISA. AB - An enzyme-linked immunosorbent assay (ELISA) to detect antigens of Giardia duodenalis in faeces was evaluated as a diagnostic tool by testing faecal samples collected during a cohort study of 229 infants living in an urban slum in Dhaka, Bangladesh. Faecal samples had been collected at enrollment, on a routine monthly basis, and repeatedly during episodes of diarrhoea and infection with Giardia, and a portion of all samples was frozen in saline. A direct smear of all had been examined by microscopy and again after concentrating cysts by ether sedimentation. A total of 2121 of the 4936 samples stored during the 22 months study were tested by the ELISA. After excluding non-specific binding, the sensitivity of the assay was 94.2% and the specificity was 98%. The presence of other parasites, including flagellated protozoa, was not linked to false positive ELISA results. There was a correlation between the number of Giardia cysts present and the ELISA optical density. Assuming that the ELISA is 100% sensitive, microscopy detected 92.4% of the infections detected by the ELISA. PMID- 8249074 TI - Serological diagnosis of Oesophagostomum infections. AB - A sensitive and specific enzyme-linked immunosorbent assay (ELISA) is described to diagnose human infection with Oesophagostomum bifurcum. In an ELISA using crude soluble antigen, prepared from adult O. bifurcum, many cross reactions occurred when measuring IgG titres in patients with other helminth infections. An ELISA based on the detection of specific IgG4, however, had a specificity of over 95%. The sensitivity of the IgG4 ELISA was difficult to assess because a reliable parasitological diagnosis is not available. The IgG4-ELISA described seems to be a powerful new tool to study the distribution of this little known but locally very common nematode parasite. PMID- 8249075 TI - Relation of the stage of parasite development in the peripheral blood to prognosis in severe falciparum malaria. AB - Admission blood films from 72 patients who died of severe falciparum malaria (50 Thai adults, 22 Gambian children) were matched retrospectively for parasitaemia with equal numbers of survivors. The peripheral blood parasites from fatal cases were more mature than those from survivors. Tiny rings (TR) comprised > 50% of parasites in 47/72 (65%) survivors but only 12/72 (17%) of fatal cases (P < 0.001). Parasites containing visible pigment (MTS: mature trophozoites and schizonts) comprised < 20% of the total parasite count in 10/72 (14%) survivors compared with 31/72 (43%) fatal cases (P < 0.001). Of the 39 patients with > 10(4) MTS/microL, 30 (81%) died. These findings were confirmed in a prospective study of 279 adult Thai patients admitted sequentially with acute falciparum malaria. Only 4 of the 19 fatal cases (21%) had > 50% TR, compared with 130 of 260 (50%) survivors, whereas > 20% MTS were found in 10/19 (53%) fatal cases, compared with 28/108 (27%) severe malaria survivors, and 26/155 (17%) patients with moderately severe malaria (P = 0.001). As a predictor of fatal outcome, the finding of either > 10(4) MTS/microL or > 5 x 10(5) parasites/microL in severe malaria had a sensitivity of 90% (95% confidence interval [CI] = 75-97%) and a specificity of 72% (95% CI = 59-86%). These observations are consistent with the hypothesis that a predominance of mature parasites in the peripheral blood reflects a greater sequestered biomass, and thus more severe disease. Simple microscopical assessment of parasite maturity on an admission blood slide provides important pathophysiological and prognostic information in severe falciparum malaria. PMID- 8249076 TI - The clinical and immunological spectrum of American cutaneous leishmaniasis. AB - American cutaneous leishmaniasis is characterized by a spectrum of clinical manifestations. These include localized, often self-healing single lesions, intermediate forms which frequently produce mucosal lesions and often show exaggerated delayed-type hypersensitivity (DTH), and the rare diffuse cutaneous leishmaniasis in which no reaction of protective cell-mediated immunity or DTH can be demonstrated. Clinical, pathological and immunological studies have begun to unravel some of the mechanisms associated with different disease manifestations, dependent on complex interactions between the host immune response, measured in terms of indices including lymphocyte subsets and lymphokines in vitro and within active lesions, and different species of Leishmania. PMID- 8249077 TI - Ischaemic colitis (necrotizing colitis, pseudomembranous colitis) in acute schistosomiasis mansoni: report of two cases. AB - Two cases of ischaemic necrosis of the sigmoid colon (necrotizing colitis) are reported in 2 brothers aged 7 and 4 years, diagnosed within a 10 d interval. The children had bathed in streams suspected to be contaminated by Schistosoma mansoni about 50-60 d before the onset of acute disease. Both patients had been previously exposed to schistosome-infected streams without showing signs or symptoms of infection. Before admission, S. mansoni eggs had not been found in the stool. Both patients presented with an apparently identical, relatively symptomatic clinical course with rapid evolution to an acute abdomen. Laparotomy disclosed, in both patients, extensive necrosis (ischaemic necrotizing colitis of schistosomal aetiology) of about 20 cm in the first child and 8 cm in the second, extending from part of the descending colon to the sigmoid. The patients were successfully operated upon (hemicolectomy plus colostomy). The histopathological findings were similar in both patients. Ischaemic necrosis with complete destruction of the mucosa and part of the submucosa was detected in the first case; in the necrotic areas a few eggs of S. mansoni were seen, with no granulomatous reaction, but surrounded by cell shadows, pycnotic nuclei and amorphous material. Necrosis extended to the muscular layer and serosa, in which schistosome granulomas in the necrotic-exudative phase were seen, as well as diffuse granulocytic exudate and fibrin. Sections of tissue from both patients contained numerous eggs and granulomas all in the same exudative phase in regional lymph nodes and near the thrombotic vessels. After surgery, the 2 patients progressed similarly. About 10 d after hospital discharge, the patients received anti-schistosomal treatment with oxamniquine. No further sign of infection was detected at subsequent recall visits. PMID- 8249078 TI - Possible clinical infection with Edge Hill virus. PMID- 8249079 TI - Drug response and genetic characterization of Plasmodium falciparum clones recently isolated from a Sudanese village. AB - We have isolated 20 clones of Plasmodium falciparum from isolates from patients attending a village clinic in Sudan during 10 d in October-November 1989. The clones were genetically diverse, having highly variable molecular karyotypes and a wide range of drug responses. Chloroquine-sensitive (50% inhibitory concentration [IC50] in the 4-15 nM range) and chloroquine-resistant clones (IC50 in the 40-95 nM range) co-existed in the population, but no obvious amplification of the P-glycoprotein homologue gene, Pgh1 (previously known as the multi-drug resistance gene, mdr1) marked the chloroquine-resistant clones. Chloroquine resistance was reversible by verapamil in these clones, although they varied in their susceptibility to verapamil alone. These observations indicate that the biochemical characteristics of the Sudanese chloroquine-resistant P. falciparum are similar to those reported from south-east Asian and Latin American isolates, which is consistent with there being a similar molecular basis for this phenomenon. PMID- 8249080 TI - Steady state pharmacokinetics of mefloquine in long-term travellers. AB - Mefloquine is an antimalarial drug with a 3-week elimination half-life, which has led to concerns that toxic accumulation may occur during weekly administration for long-term malaria chemoprophylaxis. Despite the endorsement of weekly mefloquine by the World Health Organization and the United States Centers for Disease Control, mefloquine pharmacokinetics have been incompletely studied in subjects taking the drug once weekly for more than 4 weeks. Our objective was to study plasma mefloquine concentrations in travelers taking mefloquine 250 mg once weekly for 3 months. Multiple mefloquine concentrations were measured by high pressure liquid chromatography following the 1st, 2nd and 10th to 13th of 13 weekly doses of 250 mg mefloquine taken by 15 Canadian travellers (median age 23 years; 6 male, 14 white). Steady state was achieved in all subjects by or before the 10th dose. Mefloquine pharmacokinetic values were comparable to those previously reported by other investigators. In 7 subjects, 2 peaks of mefloquine and metabolite concentration followed ingestion, suggesting redistribution of mefloquine. Mefloquine concentration 14 d after the last dose was 74% of the level 7 d after the last dose. In conclusion, pharmacokinetic values determined by this study support mefloquine weekly dosing for long-term malaria chemoprophylaxis; toxic accumulation does not occur and weekly dosing is associated with significantly higher trough levels than 14 d dosing. PMID- 8249081 TI - Neuropsychiatric side effects of mefloquine in Africans. PMID- 8249082 TI - Resistance of Plasmodium falciparum to amodiaquine in Papua New Guinea. AB - The sensitivity of Plasmodium falciparum to amodiaquine was assessed in children in the Madang region of Papua New Guinea. Fifty-four tests in vivo were carried out and 19 (35%) of these showed resistance at the RII or RIII level. Twenty-two of the 43 isolates tested showed some degree of resistance to amodiaquine in vitro. A negative correlation between the age of the patient and apparent resistance of the parasite in vivo was observed. There was no correlation between resistance in vivo and in vitro. PMID- 8249083 TI - A new micronized formulation of halofantrine hydrochloride in the treatment of acute Plasmodium falciparum malaria. AB - The current formulation of halofantrine hydrochloride has poor absorption and bioavailability. A newly developed micronized formulation was evaluated for efficacy, safety and tolerance in the treatment of acute Plasmodium falciparum malaria. The study was conducted at a plantation hospital in northern Tanzania, where chloroquine resistance is common. Sixty in-patients with mild or moderate malaria were treated with 375-750 mg micronized halofantrine hydrochloride given in 3 equal doses, 6 h apart. Patients were followed up for 28 d after therapy. Treatment was associated with rapid parasite clearance (mean clearance time = 34.8 h), fever clearance (mean time = 20 h), and clinical improvement (70% of patients were free of all presenting symptoms within 2 d). The formulation was well tolerated clinically, although 3 patients (5%) developed mild pruritus after treatment which may have been drug-related. Haematological and biochemical studies did not indicate any significant toxicity. One patient, whose immediate recovery was uneventful, was found to have a headache and low parasitaemia 3 weeks after treatment. He was readmitted to the study and treated as before. Parasitaemia, fever and headache cleared rapidly and he remained aparasitaemic for 28 d. PMID- 8249084 TI - Activity in vitro of doxycycline against multidrug-resistant Plasmodium falciparum. PMID- 8249085 TI - Anti-malaria activity of Nigerian neem leaves. PMID- 8249086 TI - Intramuscular sulfadoxine-pyrimethamine in uncomplicated chloroquine-resistant falciparum malaria during pregnancy. PMID- 8249087 TI - Eflornithine concentrations in serum and cerebrospinal fluid of 63 patients treated for Trypanosoma brucei gambiense sleeping sickness. AB - Eflornithine (difluoromethylornithine, DFMO) has recently been approved for the treatment of Trypanosoma brucei gambiense trypanosomiasis. Treatment failures have been infrequent but have occurred among patients treated with oral DFMO only, and among children. To investigate the higher frequency of failures observed in young patients, DFMO trough concentrations in serum and cerebrospinal fluid (CSF) were measured at the end of treatment in 13 children and 50 adults who had received 200 mg/kg intravenously every 12 h for 14 d. Mean DFMO concentration in CSF was significantly lower among children aged less than 12 years when compared to older patients (25.1 vs 68.9 nmol/mL, P < 0.001). Mean serum concentration was also lower in children (49.2 vs 87.5 nmol/mL, P = 0.03). Among patients who received DFMO as initial therapy for sleeping sickness, the mean CSF/serum ratio was lower in children (0.41 vs 0.91, P < 0.005). The 3 patients who failed DFMO treatment had CSF trough concentrations around or below 50 nmol/mL. Convulsions and anaemia were associated with higher drug levels and previous therapy with melarsoprol. The lower CSF drug concentrations observed in children could result from higher renal clearance and different CSF pharmacokinetics of DFMO in that age group. To avoid treatment failures, a 6 hourly regimen as well as higher DFMO dosage based on body surface area rather than on weight are recommended for children. PMID- 8249088 TI - Efficacy and tolerability of liposomal amphotericin B in Italian infants with visceral leishmaniasis. PMID- 8249089 TI - Microfilariae recurrence in Polynesian Wuchereria bancrofti carriers treated with repeated single doses of 100 micrograms/kg of ivermectin. AB - Forty-six Polynesian carriers of Wuchereria bancrofti were treated with 3 successive single doses of ivermectin, 100 micrograms/kg, given every 6 months. Immediate microfilaricidal activity of ivermectin was excellent in all carriers, since residual mean microfilaraemia levels, 2 d after each of the 3 treatments, were less than 1% of pretreatment levels. Before initial treatment, geometric mean microfilaraemia was 500 microfilaria (mf)/ml for the whole group (range 21 6398 mf/ml); 6 months after each successive treatment it was 197, 108 and 87 mg/ml, respectively, 39.4, 21.6 and 17.4% of the pre-initial treatment level. By considering the mean percentage recurrent level at 6 months after the 3rd treatment (36.8%) as a threshold, it was possible to classify the carriers into 2 groups: 17 in whom the percentage recurrent level was > 36.8% and who were considered as 'fast repopulating' individuals, and the remaining 29 who were considered as 'slow repopulating' individuals. In the latter group, 6 months after each of the 3 treatments, the recurrent microfilaremia levels were 22.7%, 8.0% and 4.9% of the pre-initial treatment level, respectively, while they were 95.1%, > 100% and > 100% in the former. The constant recurrence of mf suggests that ivermectin, at a dosage of 100 micrograms/kg, had no effect on adult worms in 'fast repopulating' individuals, whereas the progressive lessening in recurrence of mf suggests some activity (sterilizing or killing) of ivermectin on W. bancrofti macrofilariae in 'slow repopulating' individuals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249090 TI - Cerebral alveolar echinococcosis treated with albendazole. PMID- 8249091 TI - Single dose treatment with praziquantel for human Diphyllobothrium nihonkaiense infections. AB - Diphyllobothrium nihonkaiense is a large pseudophyllid cestode and one of the commonest parasites derived from fish in Japan. Three Japanese patients infected with D. nihonkaiense were treated with a single oral dose of 7, 23 or 25 mg/kg of praziquantel. All patients expelled the tapeworm with scolex, and no side effect was noted. This study indicates that single-dose treatment with praziquantel is effective against D. nihonkaiense infection. PMID- 8249092 TI - Antigenic analysis of Leishmania isolates from Tachira state, Venezuela. AB - Studies of cutaneous leishmaniasis in 3 endemic foci in Tachira state, western Venezuela have revealed sympatric populations of parasites causing both cutaneous and mucocutaneous disease. Immunological techniques and measurement of protease/acid phosphatase activities have been used to detect species-specific parasite antigens from 3 isolates from Tachira. Identified antigens of particular interest had molecular masses of 100, 82, 66, 50 and 27 kDa, but there was a high degree of heterogeneity between the antigens of the Tachira isolates and other Venezuelan strains of Leishmania braziliensis and L. mexicana. This heterogeneity has implications concerning the selection of antigens for use in serodiagnosis of leishmaniasis. PMID- 8249093 TI - Entamoeba histolytica: identification of a major surface antigen in liver abscess pus using a monoclonal antibody. PMID- 8249094 TI - Isozymic data question the specific status of some blood-sucking bugs of the genus Rhodnius, vectors of Chagas disease. PMID- 8249095 TI - Adherence of Entamoeba histolytica to hydrophobic matrices: a simple method for measuring cell surface hydrophobicity. PMID- 8249096 TI - Ubombo and the site of David Bruce's discovery of Trypanosoma brucei. AB - The site and remains believed to be those of the camp where Sir David Bruce and his wife Mary worked between 1894 and 1897, and where Bruce discovered the causative agent of nagana and its transmission by the tsetse fly, have recently been discovered at the small village of Ubombo in northern KwaZulu (Zululand), South Africa. The site where these remnants were found fits the meagre, albeit significant, information presented by Bruce in his writings on the location of the camp. PMID- 8249097 TI - A lens too far? PMID- 8249098 TI - Apparent absence of Sarcocystis infection in human tongue and diaphragm in Northern Ireland. PMID- 8249099 TI - Rejection in single versus combined pancreas and kidney transplantation in pigs. AB - Clinically, the incidence of reversible renal allograft rejection episodes appears to be higher in recipients of simultaneous pancreas/kidney (SPK) than of kidney transplantation alone (KTA); conversely, the rate of irreversible pancreas allograft rejection appears to be higher in pancreas transplant alone (PTA) than SPK recipients. Clinical/histological correlation of graft rejection in these three groups has not been precise. Therefore, we studied the incidence and histological severity of the rejection process in a large animal (pig) model of SPK (n = 36), PTA (n = 31), and KTA (n = 36) allotransplantation. SPK and PTA recipients were made diabetic pretransplant by streptozotocin (150 mg/kg). Pancreas graft exocrine secretions were bladder-drained via a duodenocystostomy for urine amylase (UA) monitoring; endocrine function was monitored by plasma glucose (PG) levels. SPK and KTA recipients underwent native nephrectomy, and renal allograft function was monitored by serum creatinine (CR). Cyclosporine, azathioprine, and prednisone were given in tapering doses from the time of transplantation. Grafts were biopsied weekly to grade histologic severity of interstitial and vascular rejection on light microscopy (LM) and for intensity of T cell infiltration on immunofluorescence. Pancreas graft exocrine function (UA above pretransplant baseline), present in 62% of PTA and 68% of SPK recipients at one week, persisted in only 7% of PTA vs. 64% of SPK pigs at 2 weeks (P = 0.0004). Likewise, pancreas graft endocrine function (PG < 200 mg/dl off insulin) was sustained longer in SPK than PTA recipients (100% vs. 84% at 1 and 91% vs. 27% at 2 weeks; P = 0.0006). However, renal allograft functional survival (serum creatinine < 3.0 mg/dl) was not significantly different (P = 0.471) between SPK and KTA recipients (36% vs. 30% at 1 and 23% vs. 13% at 2 weeks). Graft functional parameters partially correlated with biopsy observations. Pancreas allograft biopsies showed a significantly (P = 0.03 at 1 and P = 0.05 at 2 weeks) lower incidence of moderate/severe interstitial rejection in SPK than PTA recipients (67% vs. 95% at 1 and 57% vs. 92% at 2 weeks); rejection was absent in 8% of SPK and in no PTA biopsies at 1 week. Vascular rejection was moderate/severe in significantly fewer (P = 0.0013 at 1 and P = 0.023 at 2 weeks) SPK than PTA pancreas grafts (13% vs. 37% at 1 and 14% vs. 38% at 2 weeks).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8249100 TI - Long-term survival of DLA-matched segmental small-bowel allografts in dogs. AB - The aim of this study was to investigate the combined effect of DLA matching and immunosuppressive therapy on the survival of segmental small-bowel allografts in dogs. Orthotopic segmental small-bowel transplantations (25 to 30% of total small bowel length) were performed in two stages: first a heterotopic segmental small bowel transplantation, followed after 5 to 8 weeks by a second-stage operation during which the heterotopic graft was placed in an orthotopic position and the native small bowel was resected. All dogs received cyclosporine immunosuppression. Control dogs (n = 4), subjected to total enterectomy, survived 37.3 +/- 7.1 days (mean +/- SEM). Recipients of DLA-mismatched small bowel grafts (n = 6) survived 113.2 +/- 37.0 days, which was a significantly shorter time than dogs with a DLA-matched graft (n = 6, 211.5 +/- 38.8 days, P < 0.05). None of the matched allografts was rejected during CsA treatment, whereas four of six mismatched grafts were (P < 0.05). The control dogs uniformly showed progressive weight loss, steatorrhea, and hypoalbuminemia. The dogs with DLA-mismatched grafts did not regain initial body weight, whereas animals with DLA-matched grafts recovered preoperative weight after 20 weeks. Both transplanted groups showed near-normal fecal fat excretions and constant serum albumin, cholesterol, and triglyceride levels, whereas serum total protein levels increased during follow-up. We conclude that segmental small bowel transplantation between DLA matched donor-recipient pairs results in long-term survivors with an adequate nutritional status. This may have important implications for future living related small-bowel transplantation. PMID- 8249101 TI - Biohybrid artificial pancreas. Long-term function of discordant islet xenografts in streptozotocin diabetic rats. AB - Long-term function of canine, bovine, and porcine islet xenografts implanted in streptozotocin-induced diabetic rats has been achieved by islet encapsulation within permselective acrylic membrane chambers. Intraperitoneal implants of 1 x 10(4) (n = 11) or 2 x 10(4) (n = 2) encapsulated canine islets reversed the diabetic state of the recipients within 24 hr, with plasma glucose levels dropping from a preimplantation level of 480 +/- 26 (mean +/- SEM) to 97 +/- 4 mg/dl during the first month. Chambers from 2 of the animals were removed, bisected, and reimplanted at 1 week and 2 months; both animals reverted to hyperglycemia (glucose, > 200 mg/dl) in < 2 weeks. The remaining implants maintained function for a mean time of 138 +/- 16 days, whereas the 2 animals that received the higher islet dose maintained function for > 260 days. Membranes containing 2 x 10(4) bovine (n = 6) or porcine (n = 10) islets also normalized glucose concentrations, with plasma glucose levels dropping from 468 +/- 61 to 91 +/- 10 (bovine) and 97 +/- 11 (porcine) mg/dl during the first month (vs. 94 +/- 3 mg/dl for nondiabetic control rats). Three of the latter implants were removed at 1 month. All 3 animals promptly reverted to diabetes. The 3-, 6-, 9-, and 12 month graft survival rates for the remaining animals were 100%, 100%, 60%, and 40%, and 100%, 75%, 50%, and 25%, respectively. The transplant recipients showed an approximately 38-54% gain in body weight during the first 100 days after implantation, compared with < 1% (P < 0.001) and 86% (P < 0.001) for the untreated diabetic (n = 5) and normal control (n = 6) groups. Immunohistochemical staining of long-term grafts (1-20 months) revealed varying degrees of alpha-, beta-, and delta-cell granulation; the external membrane surfaces were generally free of fibrotic overgrowth and exhibited only occasional host cell adherence. Despite a problem of membrane breakage in long-term implants, these results suggest that prolonged survival of discordant transplants of porcine, bovine, and canine islets in diabetic rats can be achieved without immunosuppression. PMID- 8249102 TI - The effect of rejection and graft-versus-host disease on small intestinal microflora and bacterial translocation after rat small bowel transplantation. AB - Bacterial translocation and the development of sepsis after small bowel transplantation may be promoted by immunological damage to the intestinal mucosa or by quantitative and qualitative changes in intestinal microflora. This study assessed the effects of rejection, graft-versus-host disease (GVHD) and immunosuppression on intestinal microflora and bacterial translocation after heterotopic rat small bowel transplantation. Isografts, allografts with and without CsA immunosuppression, and the semi-allogeneic parent to the F1 hybrid GVHD model were studied. Intestinal microflora in graft and host loops and bacterial translocation to host organs and the graft mesenteric lymph node were determined. Bacterial colonies were counted and individual colonies identified using API 20E nutrient and fermentation indicator techniques. Colony counts in isografts and allografts were significantly higher than in the native intestine, whereas there was a massive overgrowth in the native intestine in the GVHD group. The species profile for the host and graft loops was similar in animals that had received isografts, allografted animals receiving CsA, and animals undergoing GVHD. However, there was a large increase in Staphylococcus epidermidis in animals with rejection. Bacterial translocation was not detected in isografted animals, but was observed in all other animal groups, with S. epidermidis being the most prevalent organism. These findings demonstrate that rejection and GVHD are associated with shifts in intestinal microflora toward potentially pathogenic organisms and that bacterial translocation into recipient tissues poses a major threat for the development of sepsis. PMID- 8249103 TI - Hepatic microcirculatory changes after reperfusion in fatty and normal liver transplantation in the rat. AB - The hepatic microcirculation in fatty and normal liver grafts in ACI rats was investigated using in vivo microscopy. Six groups were studied. They were: normal and fatty control livers (sham operated), 6-hr cold University of Wisconsin solution (UW)-preserved fatty and normal liver grafts (survival conditions, fatty and normal liver grafts), 18-hr cold UW-preserved fatty livers (nonsurvival conditions, fatty liver graft), and 24-hr cold UW-preserved normal livers (nonsurvival conditions, normal liver grafts). Fatty livers in all groups were found to have narrow and irregular sinusoids with blood cell adhesions to endothelial cells. The number of adhesions increased as the preservation time increased. Sinusoidal blood flow area decreased as the preservation time increased and was correlated with survival in both normal and fatty liver grafts. The phagocytic activity of Kupffer cells (corrected for flow) increased as the preservation time increased. The phagocytic Kupffer cell activity of the 18-hr preserved fatty liver group was greater than the activity of any other group. These features may cause liver cell death and contribute to primary graft nonfunction after transplantation of a fatty liver. PMID- 8249104 TI - The effect of fasting and exogenous adenosine on ATP tissue concentration and viability of canine pancreas grafts during preservation by the two-layer method. AB - We have demonstrated a direct correlation between a high ATP level in a canine pancreas graft after preservation by the two-layer method and good posttransplant outcome. The purpose of this study was to examine the effect of fasting and exogenous adenosine on the ATP tissue level and viability of the canine pancreas graft during preservation by the two-layer method. Graft viability was judged by graft survival following autotransplantation. Maintenance of normoglycemia for 5 days post-transplant was considered graft survival. The pancreas was harvested from either 72-hr-fated (n = 3) or -fed dogs (n = 4) and preserved by the two layer (Euro-Collins' solution [EC]/perfluorochemical [PFC]) method for 24 hr. Graft survival rates in fed and fasted groups were 4/4 (100%) and 3/3 (100%), respectively. There was no significant difference in ATP tissue concentrations between the two groups (7.48 +/- 0.55 vs. 7.03 +/- 0.74 mumol/g dry wt, NS). The pancreas was subjected to 60 min warm ischemia and then was preserved by the two layer method using EC or EC containing 5 mM adenosine for 24 hr. Without adenosine, graft survival rate was 0/3 (0%) and ATP tissue levels were not changed during preservation (1.62 +/- 0.26 vs. 1.56 +/- 0.40 mumol/g dry wt, NS). However, provision of adenosine to the graft during preservation led to the restoration of ATP tissue levels (1.90 +/- 0.54 vs. 8.13 +/- 0.98 mumol/g dry wt, P < 0.01) in 4 of 5 grafts, and these grafts functioned immediately and maintained normoglycemia after transplantation. Graft survival rate was 4/5 (80%). One of 5 grafts, however, did not survive, and the ATP tissue level was not adequately recovered during preservation compared with viable grafts (3.67 vs. 8.13 +/- 0.98 mumol/g dry wt). This study clearly demonstrates that the nutritional state of the donor has no influence on the ATP tissue level and viability of the graft during 24-hr preservation by the two-layer method. On the other hand, provision of adenosine to the graft during preservation stimulates ATP synthesis and improves the viability of the ischemically damaged pancreas. PMID- 8249105 TI - Successful extended preservation of ischemically damaged pancreas by the two layer (University of Wisconsin solution/perfluorochemical) cold storage method. AB - We have demonstrated that a two-layer (University of Wisconsin solution [UW]/perfluorochemical [PFC]) cold storage method restores the function of ischemically damaged pancreas during 24-hr preservation in canine autotransplantation model. The purpose of this study was to examine the possibility of a long-term preservation of the ischemically damaged pancreas by the two-layer (UW/PFC) method. After 60 or 90 min of warm ischemic time, pancreas grafts were preserved by the two-layer (UW/PFC) method or a simple cold storage in UW alone for up to 96 hr. A K value of i.v. glucose tolerance test more than 1.0 2 weeks after autotransplantation was considered successful preservation. After 60 min warm ischemia, limitation of preservation time by the simple cold storage in UW was 24 hr (5/5 100% and 0/5 0%; 24- and 48-hr preservation, respectively). However, the two-layer method made it possible to extend the preservation time up to 48 hr (5/5 100%, 5/5 100%, 2/5 40%, and 0/5 0%; 24-, 48-, 72-, and 96-hr preservation, respectively). After 90 min warm ischemia, the simple cold storage in UW was not effective even for 24-hr preservation (0/5 0%). However, 48-hr preservation was successful by the two-layer (UW/PFC) method (5/5 100%, 5/5 100%, and 0/5 0%; 24-, 48-, and 72-hr-preservation, respectively). After preservation by the two-layer (UW/PFC) method, ATP tissue concentrations of viable grafts were significantly higher compared with nonviable grafts (9.11 +/- 3.05 (n = 22) versus 5.22 +/- 1.02 (n = 13) mumol/g dry wt, P < 0.001). Based on analysis of individual ATP for each graft, if an ATP concentration of 6.0 mumol/g dry weight was determined as a critical value for doing the transplant, sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 84.6%, 91.7%, and 94.3%, respectively. This study clearly demonstrated that 48-hr preservation of the canine pancreas subjected to either 60 or 90 min warm ischemia was successfully achieved by the two-layer (UW/PFC) cold storage method, and ATP tissue concentration at the end of preservation by this method would predict the post-transplant outcome of the ischemically damaged pancreas just prior to transplantation. PMID- 8249106 TI - Increased susceptibility for CsA-induced hepatotoxicity in kidney graft recipients with chronic viral hepatitis C. AB - CsA-induced hepatotoxicity is a rare disorder in renal transplant recipients when low doses are administered and whole blood trough levels of CsA are regularly monitored. However, there is controversy about the clinical value of measuring CsA-metabolites, whose contribution to immunosuppression and toxicity is not fully understood. To assess the relation between low-dose CsA therapy and hepatotoxicity, we studied 128 renal transplant recipients attending our nephrology clinic. Eight of these patients had markedly elevated liver function tests. Three patients while receiving very low doses of oral CsA (< 3.8 mg/kg of body weight) presented marked derangements of CsA metabolism with abnormally increased CsA-metabolite levels. Parent drug levels were in the normal range. All 3 patients had chronic infection with hepatitis C virus and revealed histomorphologic evidence of hepatotoxicity. Hepatic dysfunction normalized when CsA was withdrawn or reduced by 50%. It is likely that hepatitis C virus infection interferes with CsA metabolism and/or biliary CsA-excretion and thus is responsible for CsA and/or metabolite-induced hepatotoxicity despite very low doses of CsA. PMID- 8249107 TI - Chronic hepatitis C after renal transplantation. Treatment with alpha-interferon. PMID- 8249108 TI - The role of the spleen in pancreas transplantation. AB - Early graft thrombosis and rejection of the graft are the two major causes of graft failure in pancreas transplantation. Inclusion of the spleen in the pancreatic graft has been purported as a possible solution to both complications, but severe graft-versus-host disease led to abolishment of this procedure. By irradiating the donor spleen ex vivo during cold storage, we successfully prevented graft-versus-host disease, allowing us to evaluate the advantages of clinical pancreaticosplenic transplantation. This study reports our experience with 12 pancreaticosplenic transplantations. Using Doppler flow measurements, we have been able to examine the hemodynamic advantages. Our results confirm the purported benefit. Vascular resistance indices in the pancreatic graft are significantly lower when the donor spleen is included. This, however, did not lower the incidence of thrombosis (2 out of 12 cases) in our study. Serial radionuclide studies with 99mTc-hexamethyl propylene amine oxime were performed for further evaluation of graft perfusion. With time the spleen uptake diminishes, compatible with atrophy of the organ. This was confirmed histologically. No indication of an immunologic advantage of transplanting the pancreas together with the spleen was found. All patients went through severe rejection crises. A transient reduction in platelet count (55-88%, mean 71%) of preoperative values was observed. This platelet drop is not seen in patients with a pancreas without spleen transplantation. We conclude that in pancreas transplantation, inclusion of the irradiated spleen has no obvious advantages for early graft thrombosis and rejection of the graft. PMID- 8249109 TI - Soluble CD23 as a sensitive marker for Epstein-Barr virus-related disorders after liver transplantation. AB - Plasma levels of the soluble fragments of Fc epsilon RII/CD23 (sCD23/IgE-binding factor) were measured to assess the level of activation of B lymphocytes associated with Epstein-Barr virus infection in 28 patients who received living related liver transplantation and were treated with FK506 and steroids. In 6 patients with symptoms of EBV infection (EBV-related disorders), the plasma concentration of sCD23 increased to more than 9.8 ng/ml at the onset of symptoms. In a patient with B cell lymphoma, the plasma levels of sCD23 increased significantly when peripheral lymphadenopathy was noticed, and remained more than 10 ng/ml during the terminal period. In 4 of 6 patients, the increase of plasma levels of sCD23 preceded the increase of anti-EBV capsid antigen IgM. In the other 2 of 6 patients, there was no significant increase of the antibody, despite the integration of EBV DNA in the mononuclear cells in their ascites. The plasma levels of sCD23 of the patients without symptoms of EBV infection did not exceed 7.5 ng/ml. In contrast, the proportion of CD20+/CD23+ B lymphocytes in peripheral blood mononuclear cells was not significantly different in the patients with EBV related disorders and those with latent asymptomatic EBV infection. Therefore, the plasma level of sCD23 is a sensitive and useful marker of EBV-related polyclonal and/or monoclonal B cell proliferation in transplanted patients with immunosuppression. PMID- 8249110 TI - Interferon-alpha and donor buffy coat transfusions for treatment of relapsed chronic myeloid leukemia after allogeneic bone marrow transplantation. AB - Eight patients with chronic myeloid leukemia relapse after allogeneic BMT were treated with IFN-alpha and buffy coat transfusions (BC) of the bone marrow donor. The antileukemic effect of this treatment was directly demonstrated in 4 patients by the disappearance of Philadelphia chromosome-positive metaphases or the loss of detectable BCR-ABL transcripts by polymerase chain reaction. In 2 patients in whom cytogenetic or polymerase chain reaction analysis was not performed, a change in hemopoietic chimerism with recurrence of donor-type hemopoiesis was demonstrated. Two patients, both treated in advanced stages of hematological relapse after BMT, did not respond. However, severe side effects of the treatment were observed: graft-versus-host disease (GVHD) occurred in 5 patients. Two of these patients progressed to severe chronic GVHD and 1 patient ultimately died of this complication. GVHD occurred in 5 of the 6 responding patients; one patient responded without developing clinical symptoms of GVHD. Six patients developed bone marrow hypoplasia after IFN/BC treatment, and pancytopenia occurred in 4 patients. None of these 4 patients recovered spontaneously and 2 patients died of complications of pancytopenia (cerebral bleeding, infection). Our results demonstrate that treatment of chronic myeloid leukemia relapse with IFN and BC transfusions is highly effective in patients with relapse in chronic phase. The occurrence of GVHD and pancytopenia, however, resulted in a high treatment associated morbidity and mortality. Whereas a response to treatment was observed in 1 patient without GVHD, indicating that GVHD and a graft-versus-leukemia effect may be clinically separable, bone marrow hypoplasia occurred in all responding patients. PMID- 8249111 TI - Islet cell hormone release immediately after human pancreatic transplantation. A marker of tissue damage associated with cold ischemia. AB - Pancreatic graft procurement, preservation, and transplantation surgery may result in damage to and loss of the integrity of endocrine cells and consequently in leakage of cell products into the insular vascular capillaries. Thus, the amount of alpha-, beta-, and pancreatic polypeptide (PP) cell products released into the vascular space of the recipient immediately after graft reperfusion may reflect islet cell injury. To test this hypothesis, we assessed glucagon, PP, C peptide, and insulin levels in a prospective study of 22 consecutive renal pancreatic transplantations. Transplantation-related parameters were used to account for differences in hormone release. Five grafts were preserved using Euro Collins preservation fluid and 17 grafts were preserved using University of Wisconsin solution (UW). The first sign of a reinstalled physiological axis was the decrease of the blood glucose concentration after a median duration of 40 min (range 5-90 min) and the association of the recipient's ambient blood glucose levels with insulin release between 25 and 180 min after reperfusion. The delay period before a fall in blood glucose was observed correlated with cold ischemia time (rs = 0.73, P < 0.001, n = 21). An immediate and marked increase in plasma levels of glucagon (from 180 +/- 18 to 585 +/- 99 ng/L, mean +/- SEM), PP (from 57 +/- 8 to 122 +/- 13 pmol/L), C-peptide (from < 0.06 +/- 0.02 to 5.43 +/- 0.63 nmol/L), and insulin (from 0.15 +/- 0.21 to 2.05 +/- 0.26 nmol/L) was observed. C peptide release correlated with glucagon (r = 0.76, P < 0.001) and PP (r = 0.60, P < 0.01). The hormone release was compared with computed tomography scans that were performed in the immediate postoperative period in 15 UW-preserved allografts. The diameter of the pancreatic head was increased and ranged from 4.5 to 7.7 cm (mean 6.2 cm). Peroperative C-peptide release significantly correlated with morphological graft changes reflected by the pancreatic head diameter (r = 0.58, P = 0.02). In a stepwise multiple regression analysis, cold ischemia time was a significant factor for the release of PP (r2 = 0.18, P = 0.049) and C peptide (r2 = 0.35, P = 0.004). We suggest that peroperative hormone release reflects endocrine tissue damage. Furthermore, cold ischemia time may jeopardize the pancreatic allograft after relatively short preservation times, even when UW is used. PMID- 8249112 TI - Ten years' experience of renal transplantation in children in the cyclosporine era. AB - Sixty renal transplantations were performed in 53 children, 0.4-16.0 years of age during the last 10 years. Fifty-five percent of the children were < or = 7 years at transplantation and 23% were < or = 2 years. Congenital nephropathies were the primary disease in 79%. Preemptive transplantation was performed in 24 first transplantations. Forty-two grafts came from living related donors and 18 came from cadaveric donors. The 1- and 5-year patient survival rates in the 0- to 7.0 year age group were 83% and 83%, respectively, and in the 7.1- to 16.0-year age group, 100% and 93%. The 1- and 5-year graft survival rates were 77% and 77% and 90% and 74% in the two groups, respectively. In children < or = 2 years old at transplantation, the 1- and 5-year patient and graft survival rates were the same, 86% and 86% in living related donors recipients, whereas they were 40% and 40% in cadaveric donors recipients. Six patients died, 3 with functioning grafts. An additional 7 grafts were lost in 6 patients, all of whom were subsequently retransplanted. The median height SD scores at transplantation was -2.98 SD in children with congenital diseases and -0.48 SD in children with acquired diseases. The median height SD scores of the 22 children followed for 3 years after transplantation was -1.06 SD. It is concluded that the survival rates obtained are satisfactory, despite the fact that the majority of the children were transplanted at a comparatively young age because of a high frequency of congenital renal disorders. PMID- 8249113 TI - Impact of innovative techniques on the waiting list and results in pediatric liver transplantation. AB - The wide application of liver transplantation in children is hampered by the shortage of size-matched pediatric donors; this results in high mortality rate on the waiting list, a long waiting time, worsening of the clinical condition of the waiting patient, deterioration of the overall results, and an increase in the cost. Reduced-size liver transplants have been shown to be a safe way to alleviate the shortage of size-matched organs. We have retrospectively analyzed the impact of the reduced-size liver transplants on the waiting list and the results in a consecutive series of 314 transplants performed in 261 children over an 8-year period (1984-1991). Among these 314 grafts, 160 (51%) were innovative techniques including 86 reduced livers (stricto senso), 66 partial livers (with preservation of the recipient vena cava), and 8 split livers. Such an extensive use of these technical variants allowed a sharp decrease in the waiting list mortality: from 14.9% between 1984 and 1989 to 6.6% in 1990 and 5% in 1991; the corresponding figures for infants registered under the age of 1 year were 25%, 13.3%, and 8.3%, respectively. Results obtained with a full-size graft or a technical variant were similar regarding surgical complications (with a significantly lower incidence of arterial thrombosis for the reduced transplants), graft loss, and patient survival. The 5-year survival of the whole group was 78.1% without any significant difference regarding type of transplant, indications (with the best results: 89.4% 5-year survival obtained in 41 children grafted for metabolic diseases), or age (the 5-year survival was 82.2% for the 41 infants transplanted under the age of 1 year, 78.9% for the 124 children transplanted between 1 and 3 years, and 81.3% for the 96 children transplanted between 6 and 15 years). This series of reduced-size liver transplants, which is the largest worldwide single institutional experience, confirms that the extensive use of reduced transplants in children is safe; this study also shows that innovative techniques, including the split liver, allow a drastic decrease of the waiting list mortality of candidates in the pediatric age range without alterations of the results. PMID- 8249114 TI - The efficacy and toxicity of rapamycin in murine islet transplantation. In vitro and in vivo studies. AB - We performed an in vivo and in vitro dose-response study of the novel immunosuppressive macrolide antibiotic rapamycin looking at murine islet allograft survival, impact on glucose homeostasis, and possible tissue toxicity. A total of 300 islets were isolated from CBA/J mice (H-2k) and transplanted beneath the renal capsule of streptozotocin-induced diabetic BALB/c (H-2d) recipients. Seven groups of allografted mice received intraperitoneally for 7 days post-transplant: no immunosuppression (n = 8); vehicle only (carboxymethyl cellulose) (n = 6); or rapamycin at dosages of 0.05 (n = 8), 0.1 (n = 8), 0.3 (n = 8), 1.0 (n = 8), or 5.0 (n = 6) mg/kg/day. Blood glucose was monitored on alternate days, with graft failure defined by the first day of persistently high blood glucose (> 14 mmol/L). The 0.1 and 0.3 mg/kg/day groups showed statistically significant prolongation of diet allograft survival (P < 0.01) when compared to the controls and vehicle-treated mice. Three mice in both the 0.1 and 0.3 mg/kg/day groups and one mouse in the 0.05 mg group reached 100 days normoglycemia and, following nephrectomy of the islet-bearing kidney, returned to hyperglycemia. The 0.05, 1.0, and 5.0 mg/kg/day groups showed no statistically significant prolongation of graft survival. In addition, the higher dosage (1.0 and 5.0 mg/kg/day) groups had erratic blood glucose control. Histologically, there was no evidence of toxicity seen in any of the multiple organ samples. In the in vitro analysis, BALB/c (H-2k) islets cultured in either 0, 10, 30, or 100 ng/ml rapamycin had no significant differences in insulin secretion following a 24-hr culture period; however, there was a significant deterioration in glucose stimulated insulin release after 72 hr culture at high rapamycin concentration (100 ng/ml). Rapamycin significantly prolonged murine islet allograft survival. At doses 10 to 50 times the effective antirejection dosage, we demonstrated adverse impact on glucose homeostasis without histological evidence of end-organ toxicity. We also demonstrated an adverse impact on insulin release in vitro following prolonged culture (72 hr) in a high concentration of rapamycin. PMID- 8249115 TI - Induction of specific unresponsiveness to rat islet allografts by intrathymic UVB donor spleen cells. AB - Recently, we showed that intrathymic (i.t.) injection of UVB-irradiated (600 J/m2) spleen cells induces donor-specific unresponsiveness to cardiac allografts in the sublethally irradiated (200 rads, TBI) recipients in the Lewis-to-ACI rat combination. This study examined if i.t. injection of UVB donor SC could induce specific unresponsiveness to neovascularized (islet) allografts in the same rat combination of Lewis-to-ACI. Streptozotocin-induced diabetic ACI rats pretreated with sublethal TBI (200 rads) 7 days prior to intraportal transplantation of freshly isolated Lewis islets reject their grafts in 10.2 +/- 2.9 days compared with rejection of islets in 8.5 +/- 2.6 days in unmodified controls. Recipient pretreatment with i.t. injection of UVB donor SC combined with sublethal TBI (200 rads) 7 days prior to islet transplantation induced indefinite graft survival (> 200 days) in 3 of 7 animals. Similar treatment failed to prevent acute rejection of third-party (WF) islets, thus demonstrating donor-specificity. Treatment with sublethal TBI (200 rads) on the day of islet transplantation led to permanent graft survival in 2 of 5 animals that received i.t. inoculation of UVB donor SC 7 days prior to islet transplantation. Conditioning of the recipients with a sublethal TBI dose of 300 rads on the day of islet transplantation, which alone delayed graft rejection for only 19 days, led to indefinite graft survival (> 150 days) in all animals pretreated with i.t. injection of UVB donor SC. Extrathymic inoculation of donor UVB SC via subcutaneous, intraperitoneal, intratesticular, and intravenous routes, respectively in similarly prepared animals failed to prolong islet survival, thus confirming the privileged position of the thymus in the induction of tolerance. Our findings suggest that this new strategy of immunomodulation with donor UVB SC is potentially useful for induction of donor specific unresponsiveness. PMID- 8249116 TI - Tolerance induction to rat islet allografts by intrathymic inoculation of donor spleen cells. AB - Intrathymic (i.t.) injection of UVB donor spleen cells induces donor-specific unresponsiveness to cardiac and islet allografts in sublethally irradiated recipients in the low-responder Lewis-to-ACI rat combination. This study examined whether unresponsiveness to islet allografts could be achieved following i.t. inoculation of untreated or UVB-irradiated donor SC under the cover of peritransplant immunosuppression with sublethal TBI or ALS in the high-responder combination of WF-to-Lewis rats. The results of this study show that i.t. injection of untreated SC combined with sublethal TBI or ALS led to permanent islet allograft survival in 50% of recipients, while i.t. injection of UVB donor SC combined with sublethal TBI or ALS resulted in indefinite graft survival in 80 100% of recipients. Third-party (BN) islets were rejected normally in this model, confirming donor-specificity of unresponsiveness. Extrathymic inoculation of UVB treated donor SC by the subcutaneous, intratesticular, or intravenous routes in similarly immunosuppressed animals did not result in any prolongation of islet allograft survival, thus confirming the importance of the thymus in induction of tolerance in this model. The unresponsive recipients that were challenged with 2nd-set allografts 100 days after islet transplantation, permanently accepted donor-type but not third-party (ACI) cardiac allografts, thus proving that recipients are indeed tolerant to donor alloantigens and that such tolerance is donor- and not organ-specific. This study proposes a novel strategy of immunomodulation that may be useful in induction of specific unresponsiveness to organ allografts. PMID- 8249117 TI - The impact of acute episodes of rejection on the generation of chronic rejection in rat renal allografts. AB - We have investigated the impact of the frequency of acute rejection episodes on the generation of chronic rejection in long-surviving rat renal allografts. A total of 33 renal transplantations was performed from DA to WF rats, receiving different cyclosporine-based immunosuppressive regimens. As a consequence, different numbers of acute rejection episodes were recorded in the recipients, all of which were successfully treated with cyclosporine. Upon sacrifice at 12 weeks posttransplantation, the frequency of acute rejection episodes was correlated with the major histological parameters of chronic rejection and with graft function. The intensity of the major histological parameters of chronic rejection, exemplified by vascular intimal proliferation and glomerular mesangial matrix increase, and the decline in graft function between the no-rejection vs. rejection groups, were directly proportional to the number of acute rejection episodes. Vascular intimal proliferation increased from 0.5 +/- 0.4 (arbitrary units) in the no-rejection group to 1.7 +/- 0.9 (P = 0.0093) after one rejection episode, to 2.2 +/- 0.3 (P = 0.0001) after two, and to 2.2 +/- 0.5 (P = 0.0014) after three or four rejection episodes. Glomerular mesangial matrix increased from 1.2 +/- 0.3 (arbitrary units) in the no-rejection group to 1.9 +/- 0.6 (P = 0.017) after one, to 2.2 +/- 0.5 (P = 0.0005) after two, and to 2.1 +/- 0.4 (P = 0.003) after three or four rejection episodes. Serum creatinine increased from 93 +/- 24 mumol/L in the no-rejection group to 196 +/- 92 mumol/L (P = 0.016) after one, to 238 +/- 38 mumol/L (P = 0.0001) after two, and to 308 +/- 85 mumol/L (P = 0.0016) after three or four rejection episodes. Prolongation of the preoperative ischemia time from 30-60 min correlated with an increase in the number of acute rejection episodes as well as an increase in chronic changes. To conclude, in the rat renal allograft model, acute allograft rejection carries a highly significant correlation with the development of chronic rejection. PMID- 8249118 TI - Alterations in renal interleukin-1 production during kidney transplant rejection in the rat. The effects of high-dose methylprednisolone. AB - To characterize the role of interleukin-1 in renal allograft rejection, we examined the temporal relationship of IL-1 production to changes in renal function and histology in a rat kidney transplantation model. In rat renal allografts, both glomerular filtration rate and renal plasma flow (RPF) fell progressively from days 4 through 6 following transplantation. The reduction in allograft function was accompanied by histologic changes consistent with rejection and enhanced steady-state levels of IL-1 beta mRNA measured by Northern blot. This increase in IL-1 beta mRNA levels was associated with a forty-fold increase in IL-1 bioactivity in eluates of kidney allografts compared with isografts. In rejecting allografts, these changes also coincided with increased production of thromboxane B2 (TxB2) by the graft. To attempt to modify IL-1 production in the transplanted kidney, a separate group of animals with renal allografts were treated with 80 mg/kg/day of methylprednisolone for 6 days. GFR in MP-treated animals was significantly preserved compared with vehicle-treated animals. However, similar histologic manifestations of rejection were found in both groups. Although IL-1 beta mRNA levels in the kidney were not changed with MP treatment, renal IL-1 bioactivity was reduced four-fold in animals that received MP compared with controls. Thus, IL-1 beta gene expression and IL-1 protein production are stimulated in rejecting kidney transplants. MP administration improves allograft function and inhibits IL-1 production, apparently at a post-transcriptional level. We hypothesize that overproduction of IL-1 during kidney transplant rejection may promote allograft dysfunction and injury. Some of the beneficial effects of corticosteroids in acute rejection may be mediated through inhibition of IL-1 release within the allograft. PMID- 8249119 TI - Long-term survival and strain-specific tolerance induction in rat-to-mouse neonatal islet xenografts. AB - These studies were designed to determine (1) if culture-isolated, neonatal rat islets are capable of inducing xenogeneic tolerance in mice and (2) whether this tolerance is species- or strain-specific. We attempted to induce xenogeneic tolerance by transplanting culture-isolated neonatal FSH islets to 26 diabetic C57B1/6 recipients. These animals received one injection of ALS at the time of transplant. Fifteen (58%) animals remained reversed by xenotransplant for > 173 days. To assess the development of strain or species-specific tolerance, 14 of the animals bearing long-term surviving FSH grafts were divided into 3 treatment groups. Animals in group 1 were nephrectomized to remove the initial graft and then retransplanted with uncultured, adult FSH islets; animals in group 2 were retransplanted with uncultured, adult FSH islets without nephrectomy; and group 3 animals were nephrectomized and retransplanted with uncultured, adult third-party islets (WF). In naive controls, adult FSH islets were rejected in 9 +/- 2 days. The MST for adult FSH grafts transplanted to nephrectomized recipients was 104 +/ 54 days, with 4 out of 5 (80%) surviving until sacrifice 90-171 days posttransplant. The MST for FSH grafts transplanted to nonnephrectomized recipients was 120 +/- 70 days with 3 out of 4 (75%) surviving until sacrifice 143-154 days posttransplant. Thus, it appears that the initial neonatal FSH transplant induced the development of immune tolerance to highly immunogenic FSH islet tissue. In contrast, the MST for third-party adult WF grafts was 27 +/- 13 days compared with an MST of 36 +/- 24 days in naive controls. Thus, it appears that the xenogeneic tolerance induced by neonatal FSH islets was strain rather than species-specific. Factors such as the close evolutionary relationship between rats and mice, the neonatal condition of the initial graft, and its relative lack of donor APCs are included in a discussion of possible mechanisms of tolerance induction. PMID- 8249120 TI - Prevention of complement-mediated activation of xenogeneic endothelial cells in an in vitro model of xenograft hyperacute rejection by C1 inhibitor. AB - The complement system plays a major role in hyperacute rejection of discordant xenografts. In an immediately vascularized xenograft of a porcine organ to a primate, natural antibodies bind to the vascular endothelium of the graft, triggering activation of complement via the classical pathway. One consequence of antibody binding and complement activation is the activation of endothelial cells leading to the loss from the cells of heparan sulfate. We explored to what extent the classical pathway regulatory protein C1 inhibitor (C1 inh) would inhibit complement-mediated cytotoxicity and activation of endothelial cells. Cultured porcine aortic endothelial cells were used as a model for a xenogeneic organ and human serum as a source of xenoreactive natural antibody and complement. Addition of purified human C1 inh to human serum inhibited deposition of C4b and iC3b and cytotoxicity after the serum was reacted with the cultured cells. C1 inh prevented, in a dose-dependent manner, activation of the endothelial cells, as manifested by release of heparan sulfate. These observations demonstrate that C1 inh added in sufficient amounts to human serum can effectively inhibit C1 activation in an antigen-antibody system. These studies extend our previous findings consistent with the concept that complement activation occurs via the classical pathway in models of hyperacute rejection in which porcine vascular endothelial cells are in contact with human serum containing xenogeneic natural antibodies against the endothelial cells. Thus, our results suggest a potential clinical use of C1 inh in conjunction with other therapies to prevent hyperacute rejection in xenogeneic combinations mediated by complement activation via the classical pathway. PMID- 8249121 TI - Iron chelation suppresses mononuclear cell activation, modifies lymphocyte migration patterns, and prolongs rat cardiac allograft survival in rats. AB - Iron influences host immunity, in part by affecting the function and migration patterns of T cell subpopulations. Removal of iron stores from the body by chelation decreases proliferation and differentiation of T cells, as shown in models of autoimmunity and pancreatic islet transplantation. We have examined the influence of iron chelation on rejection of vascularized heart allografts in rats. Two protocols were investigated: "treated" recipients received desferrithiocin (30 mg/kg/day) orally for 10 days beginning the day of transplantation; "pretreated" hosts received a similar dose of the drug for 10 days before engraftment. Graft survival increased from about 7 days in untreated animals to 14-16 days in both treatment groups (P < 0.001). Histological and immunoperoxidase studies of allografts at day 7 showed that iron chelation resulted in only a mild reduction in cell infiltration, but in a marked decrease in graft edema and interstitial hemorrhage and essentially complete suppression of mononuclear cell activation and cytokine production. Chelation therapy was also found to inhibit profoundly cytokine (TNF-alpha) production in rats treated with LPS, consistent with the effects observed in situ in the allografts. In vitro studies showed that pretreatment significantly inhibited the mixed lymphocyte reaction. Chelation also influenced migration of T lymphocyte subsets: treatment stimulated migration of CD4+ lymphocytes to peripheral lymph nodes; pretreatment strikingly and selectively increased CD8+ cell migration into parathymic lymph nodes draining the graft, with the opposite effect on nondraining node groups. We conclude that treatment with iron-chelating agents has several effects on host alloresponsiveness in a rat heart graft model secondary to inhibition of immune activation; these include prolongation of graft survival, inhibition of the mixed lymphocyte reaction (pretreatment), marked depression of cytokine production, and alteration in recirculation patterns of lymphocyte subpopulations. PMID- 8249122 TI - Exposure of rat pancreatic islets to RS-61443 inhibits beta-cell function. AB - In this study, the long-term effects of the immunosuppressive drug RS-61443 on pancreatic beta-cell function have been evaluated. For this purpose, isolated rat pancreatic islets were precultured for 6 to 7 days in RPMI 1640 medium + 10% FCS and then cultured for another 6 days in the absence or presence of RS-61443 (15, 60, 120, and 240 mg/L). The islet DNA content was decreased by 20-30% after culture at all concentrations of the drug. At the highest drug concentration, there were disarrangements in the islet architecture. This was probably not confined to only beta-cells, since the ratio of insulin to DNA was not decreased. On day 6, glucose-stimulated insulin secretion was inhibited by 75-90%, whereas rates of islet (pro)insulin biosynthesis and glucose oxidation were not or only slightly affected. After 48 hr of culture in the absence of the drug, the decrease in islet DNA content remained, but the impaired insulin secretion was partly restored. In acute in vitro experiments, RS-61443 initially enhanced glucose-stimulated insulin secretion, but during a second hour of incubation, there was an inhibition. In variance to this latter finding, islet glucose oxidation was inhibited already during the first hour. The present results suggest that long-term exposure to RS-61443 in vitro impairs beta-cell function and this might be due to an inhibition of formation of guanosine metabolites. In other experiments, rats and mice were treated for 6 days with intraperitoneal injections of RS-61443 (70 mg/kg body wt). The pancreatic insulin concentration was not changed, but during glucose tolerance tests, minor impairments were observed. In conclusion, the present data suggest that RS-61443 is potentially harmful to beta-cells. PMID- 8249123 TI - Blood distribution of rapamycin. AB - Rapamycin (RAPA) is a potent new immunosuppressive drug. Although blood concentration monitoring of RAPA is being performed in preclinical and clinical trials, little is known regarding the blood distribution of the drug. Such information would have an impact on the medium used for analysis of the drug. The distribution of RAPA was investigated by spiking human whole blood having an initial temperature of either 4 degrees C or 22 degrees C with a constant amount of 3H-RAPA and increasing amounts of RAPA to a final concentration of 5-100 micrograms/L. The drug concentration spans the range seen when immunosuppressive doses of the drug are administered. This was followed by incubation of the blood at 37 degrees C for 0 to 60 min before separation of cells. The dpm in the resulting plasma and RBC fractions was determined by scintillation counting. The plasma to formed blood elements and plasma to whole blood ratios were 0.05 +/- 0.051 and 0.09 +/- 0.016, respectively (mean +/- SD, n = 50). The distribution did not exhibit any temperature or concentration dependence. The proportion of the drug among cellular components was as follows (mean % distribution +/- SD); RBC 94.5 +/- 4.9%; plasma 3.1 +/- 2.5%; lymphocytes 1.01 +/- 1.02%; and granulocytes 1.0 +/- 0.88%. The free or unbound fraction of RAPA over the plasma concentration range of 5-100 micrograms/L as determined by ultracentrifugation was 2.5 +/- 0.2%. The drug was found to be associated primarily with nonlipoprotein fractions in plasma. The results suggest from an analytical perspective that whole blood as compared with plasma would be the most suitable medium for analysis due to the higher concentrations found in the former. PMID- 8249124 TI - Macrophage-generated nitric oxide as cytotoxic factor in destruction of alginate encapsulated islets. Protection by arginine analogs and/or coencapsulated erythrocytes. AB - Isolated rat islets were microencapsulated in alginate beads of about 1.5 mm in diameter. These were cocultured with activated or resident peritoneal macrophages of syngeneic rats for 24 hr. Examination of the encapsulated islets by transmission electron microscopy showed that the islets were lysed by activated (80.0 +/- 12.8% of islets lysed), but not by resident, macrophages (17.5 +/- 12.2% lysis) despite encapsulation. Islet lysis was inhibited in a concentration dependent manner by a specific nitric oxide-synthase inhibitor (0.5 mM NG-methyl L-arginine: 5.9 +/- 3.9% lysis) in an L-arginine-reversible manner (0.5 mM NG methyl-L-arginine + 10 mM L-arginine: 55.1 +/- 16.6% lysis). Incubation of encapsulated islets with 3 different nitric oxide-generating compounds also resulted in a concentration-dependent islet lysis. Coencapsulation of autologous erythrocytes was found to be an effective and easy way of protection from macrophage-mediated lysis. Protection was dependent upon the number of erythrocytes coencapsulated. This in vitro study demonstrates that nitric oxide secreted by activated macrophages is able to destroy islets despite encapsulation in alginate, and that both, inhibition of nitric oxide formation using enzyme inhibitors and scavenging of nitric oxide once formed exploiting the hemoglobin of autologous erythrocytes, protect encapsulated islets from destruction. PMID- 8249125 TI - Cyclosporine and cremaphor modulate von Willebrand factor release from cultured human endothelial cells. AB - Cyclosporine has been associated with microangiopathic hemolysis (MAHA) and other thrombotic complications of bone marrow and renal transplantation. MAHA is characterized by intravascular platelet aggregation, which, in some situations, is thought to be mediated by hyperactive high molecular weight von Willebrand factor (vWF). We have hypothesized that transplant-related MAHA may be caused by CsA-mediated release of von Willebrand factor from endothelial cells. This hypothesis was tested by studying vWF release from human umbilical vein endothelial cells primed with either CsA or cremophor EL. CsA and cremophor alone did not increase vWF release until toxic concentrations were reached (50-100 micrograms/ml). However, at therapeutic concentrations (0.1-5 micrograms/ml) vWF release by cells stimulated with thrombin, histamine, PMA, and the calcium ionophore A23187 was enhanced by both CsA and cremophor in a concentration dependent manner. In single isolated endothelial cells, the thrombin-induced increase in cytosolic free calcium was enhanced by both CsA and cremophor. Preincubation for 24 hr with CsA but not cremophor suppressed vWF release after thrombin stimulation. These observations were mirrored by a concentration dependent suppression of [3H]thymidine uptake by CsA. We conclude that CsA vehicle, cremophor, enhances stimulated vWF release in vitro, probably by processes dependent upon increased cytosolic free calcium. This suggests a possible mechanism for thrombotic transplant complications. PMID- 8249126 TI - The detection of cytotoxic T cells with high-affinity receptors for donor antigens in the transplanted heart as a prognostic factor for graft rejection. AB - Alloreactive T lymphocytes are the initiators and effectors of acute rejection of organ transplants, and T cells with high-affinity receptors for antigen might be especially implicated in this process. It has been shown that the cytotoxic capacity of CTL with low affinity for alloantigens can be inhibited with CD8 mAb, while high-affinity CTL are not affected. To investigate whether the presence of such high-affinity cells in human heart transplants may be predictive for acute rejection, we analyzed their frequency in cultures derived from endomyocardial biopsies in 19 patients, 9 of whom had never experienced acute rejection and 10 who had had one or more rejection episodes. IN the rejectors, already before histological signs of rejection (myocyte damage) had developed, significantly higher donor-reactive CTL frequencies were found compared with the nonrejectors (medians of 10,586 vs 1,169 reactive cells per 10(6) tested cells, P = 0.002). After CD8 inhibition, the difference between rejectors and nonrejectors was even more pronounced (P < 0.001). In patients with rejection, the number of CD8 resistant, high-affinity CTL was higher than 1000 per million cells in all cases, while in patients who had never experienced rejection this number was less than 1000. As these CTL characteristics are already present before the first histological signs of rejection have developed, this might be used as a prognostic factors. PMID- 8249127 TI - Comparison of the abilities of MHC-compatible bone marrow cells and lymph node cells to induce tolerance of skin allografts in rats. AB - A comparison has been made of the abilities of bone marrow cells and lymph node cells to induce tolerance of skin when inoculated into neonatal rats known to differ only with regard to non-MHC incompatibilities, including putative skin specific (Skn) antigens. Each recipient received 50 x 10(6) cells, and tolerance was confirmed by the permanent acceptance of donor-strain neonatal heart tissue transplanted to the pinna of the ear. In 5 of the 8 MHC-compatible strain combinations tested, BMC were significantly more effective than LNC in inducing tolerance of skin, whereas in one situation LNC proved more efficient. Although the results are in accord with the occurrence of Skn antigens in rats, it appears that at least some of the antigens involved must also be expressed by BMC or LNC, but not equally by both of these tissues. The results also provide evidence that in rats, as in mice, the MHC can play a major role in determining the response to, and/or the immunogenicity of, Skn antigens. PMID- 8249128 TI - Expression of MHC class II A alpha k transgene. Pairing with endogenous beta chain is dependent upon affinity and competition. AB - Studies on cell lines transfected with MHC class II genes have revealed important limitations on the assembly of haplotype-mismatched A alpha/A beta complexes. We have investigated pairing restriction in vivo by analyzing transgenic mice that carry an A alpha k gene on several H-2 haplotypes. Previously, we reported the A alpha k transgenic expression on the cell surface of H-2b and H-2u strains. Further transcomplementation studies performed with high A alpha k-expressing transgenic lines revealed that A alpha k chain can pair with A beta b,s,f,d,v, or u chains but not with A beta q. Homozygosity for the A alpha k transgene increased the expression of mixed heterodimers. The inefficient assembly of haplotype-mismatched class II polypeptides results from their inability to compete with the matched pairs. The higher affinity between the matched pairs can be overcome by increased synthesis of the mismatched pair. Thus, there is no "strict" sequence-based restrictions on pairing. PMID- 8249129 TI - A human lymphocytotoxic autoantibody is encoded by antibody variable region genes in their germ line configuration. AB - Lymphocytotoxic autoantibodies are commonly found in sera from kidney dialysis patients, where they interfere with interpretation of the crossmatch test. We have produced a monoclonal lymphocytotoxic autoantibody by EBV transformation of PBLs from a dialysis patient, followed by fusion of the transformed cells with a heteromyeloma. The autoantibody derived, called FWE, was IgM kappa class and its pattern of reactivity against T and B lymphocytes, cells from patients with chronic lymphocytic leukemia and K562, was similar to that described for lymphocytotoxic autoantibodies found in sera. It was absorbed by fetal but not adult human erythrocytes, suggesting the antigenic determinant might be the blood group antigen i. cDNA encoding the variable domain of FWE was amplified by polymerase chain reaction, cloned into the vector M13 mp18, and sequenced. The variable region of the kappa light chain (V kappa) was 98.8% identical over a 260 bp stretch with a known germ line sequence and the junctional (J kappa) region was identical over 16 bp with the germ line sequence J kappa 2. The variable region of the heavy chain (VH) was 99.3% identical over a 268-bp overlap with the germ line gene VH4.21, a member of the VHIV family, and the junctional region of the heavy chain (JH) was identical with the germ line JH gene JH5 over 46 bp, with a truncated 5' end. The diversity region was not identified. These data suggest that the genes required to produce human lymphocytotoxic autoantibodies are encoded within the germ line and, therefore, that all dialysis patients may be able to produce them under certain circumstances. PMID- 8249130 TI - Influence of HLA matching on rejections and short- and long-term primary cadaveric allograft survival. AB - Distribution of cadaveric donor kidneys, based upon the donor-recipient HLA match grade, remains one of the major controversies in transplantation. To determine whether matching results in fewer rejection episodes and better graft survival, we retrospectively studied our single-center patient population of 683 cyclosporine-prednisone-treated primary cadaveric renal allograft recipients. For 237 recipients of well-matched HLA A, B kidneys (< or = 2 HLA A, B mismatches [MM]) the 1-, 3-, 5-, and 7-year graft survivals of 76%, 66%, 62%, and 61%, respectively, were not significantly different from those of 71%, 65%, 63%, and 63%, respectively, for the 446 poorly matched HLA A, B (> 2 HLA A, B MM) recipients. Similarly, the 1-, 3-, 5-, and 7-year graft survivals for the 307 recipients of well-matched HLA-DR kidneys (0 or 1 DR MM) of 74%, 65%, 63%, and 61%, respectively, were not significantly different from those of 72%, 65%, 63%, and 62%, respectively, for the 366 poorly matched (2 DR MM) recipients. Patient survivals were comparable at each time point for well- vs. poorly matched recipients. Similarly, donor-recipient HLA A, B, and DR matching was not beneficial in retransplant recipients who were transplanted following negative NIH and antiglobulin (AHG) crossmatches when testing both historical (high-PRA) and pretransplant sera. Since rejection episodes may be a more sensitive indicator of immune response than graft loss, we also analyzed the relationship between donor-recipient HLA match grade and posttransplant rejections. A total of 60% (n = 413) of recipients experienced no rejections and had 1-, 3-, 5-, and 7 year graft survivals of 82%, 78%, 74%, and 73%, respectively; 32% (n = 215) of patients who experienced 1 rejection had 1-, 3-, 5-, and 7-year graft survivals of 58%, 48%, 44%, and 43%, respectively (P < 0.001 for graft survival of 0 vs. 1 rejection). The remaining 8% (n = 55) of recipients experienced more than 1 (> 1) rejection and had 1-, 3-, 5-, and 7-year graft survivals of 62%, 38%, 36%, and 36%, respectively (P < 0.001 for graft survival of 0 vs. > 1 rejection and P < 0.01 for graft survival of 1 vs. > 1 rejection). The mean numbers of rejections/patient experienced by well-matched vs. poorly matched recipients were comparable and not significantly different.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8249131 TI - A comparative study of cyclosporine and its derivative SDZ IMM-125 in canine renal allografts. AB - The immunosuppressive efficacy of CsA and its derivative SDZ IMM-125 was compared in highly mismatched mongrel dogs in receipt of renal allografts. At an equal dose of 15 mg/kg/day and in the same drug vehicle, SDZ IMM-125 was not superior to CsA in prolonging allograft survival. Whole blood levels of SDZ IMM-125 were lower than those achieved for CsA. No specific drug-related side effects were noticed in this model. PMID- 8249132 TI - Human xenoreactive natural antibodies--avidity and targets on porcine endothelial cells. PMID- 8249133 TI - Prolongation of cardiac xenograft survival in a hamster-to-rat combination by recipient treatment with deoxymethylspergualin. PMID- 8249134 TI - A new method for determining graft function after liver transplantation by near infrared spectroscopy. PMID- 8249135 TI - Functional evaluation of mouse bone marrow T lymphocytes. PMID- 8249136 TI - Coinjection of anti-interleukin-2 receptor antibody diminishes antiantibody formation during prolonged treatment with anti-CD3 antibody. PMID- 8249137 TI - The failure of donor essential fatty acid deficiency to prevent renal allograft rejection in rats. PMID- 8249138 TI - Epstein-Barr virus--associated posttransplant lymphoproliferative disorder presenting as perirenal transplant lymphocele. PMID- 8249139 TI - The effect of portal venous tolerance on the survival of small bowel allografts in the rat. PMID- 8249140 TI - Delayed-start cyclosporine immunosuppression reduces graft cell migration after small bowel transplantation. PMID- 8249141 TI - Cytomegalovirus-induced severe thrombocytopenia after liver transplantation. PMID- 8249142 TI - The first report of a successful delivery in a woman with an ABO-incompatible kidney transplantation. PMID- 8249143 TI - Prolonged venovenous bypass for intraoperative failure of transplanted livers--a report of three cases. PMID- 8249144 TI - Inflammation and hyperalgesia: highlighting the team effort. PMID- 8249145 TI - Adenosine A3 receptors: two into one won't go. PMID- 8249146 TI - In vitro approaches can predict human drug metabolism. PMID- 8249147 TI - Gaddum revisited. PMID- 8249148 TI - Constitutive activity of receptors coupled to guanine nucleotide regulatory proteins. AB - Adrenoceptors are prototypic members of the superfamily of seven transmembrane domain, G protein-coupled receptors. Study of the properties of several mutationally activated adrenoceptors is deepening understanding of the normal functioning of this ubiquitous class of receptors. The new findings suggest an expansion of the classical ternary complex model of receptor action to include an explicit isomerization of the receptors from an inactive to an active state which couples to the G protein ('allosteric ternary complex model'). This isomerization involves conformational changes which may occur spontaneously, or be induced by agonists or appropriate mutations which abrogate the normal 'constraining' function of the receptor, allowing it to 'relax' into the active conformation. Robert Lefkowitz and colleagues discuss the physiological and pathophysiological implications of these new insights into regulation of receptor activity. PMID- 8249149 TI - Molecular basis of muscarinic acetylcholine receptor function. AB - Muscarinic acetylcholine receptors play important roles in numerous physiological functions including higher cognitive processes such as memory and learning. Consistent with the well-documented pharmacological heterogeneity of muscarinic receptors, molecular cloning studies have revealed the existence of five distinct muscarinic receptor proteins (M1-M5). Structure-function relationship studies of the cloned receptors have been greatly aided by the high degree of structural homology that muscarinic receptors share with other G protein-coupled receptors. In this review, Jurgen Wess discusses recent mutagenesis studies that have considerably advanced our knowledge of the molecular details underlying muscarinic receptor function. PMID- 8249150 TI - Vineyard peptide conference bears fruit. PMID- 8249151 TI - New perspectives in osteoporosis. PMID- 8249152 TI - The 'inner core' of inwardly rectifying K+ channels. PMID- 8249153 TI - Reversible inhibitors of the gastric H+/K(+)-transporting ATPase: a new class of anti-secretory agent. PMID- 8249154 TI - Do the K+ channel openers relax smooth muscle by opening K+ channels? AB - During the past decade, a group of chemically heterogeneous compounds known as the K+ channel openers has emerged. These compounds open a certain class of K+ channels (ATP-sensitive K+ channels) in the sarcolemma of vascular smooth muscle cells, which leads to hyperpolarization of the cell membrane and relaxation of the tissue. The mechanisms by which hyperpolarization affects smooth muscle contraction and contractility can thus be examined. Hyperpolarization induced by these K+ channel openers prevents Ca2+ entry through voltage-operated Ca2+ channels. Surprisingly, and by mechanisms not yet defined, hyperpolarization of the cell also reduces agonist-induced accumulation of inositol 1,4,5 trisphosphate (and consequently, Ca2+ mobilization from intracellular stores), and the Ca2+ sensitivity of the contractile apparatus. In addition, recent evidence reviewed here by Ulrich Quast suggests that the K+ channel openers possess further mechanisms of vasorelaxation not linked to the opening of plasmalemmal K+ channels. PMID- 8249155 TI - Transforming growth factor-beta 1 and basic fibroblast growth factor in the injured CNS. AB - After injuries that penetrate the mature brain or spinal cord, damaged axons initially show a growth response, but later their regeneration is aborted as a dense permanent scar is laid down within the core of the wound. Functional recovery from such injuries is poor and morbidity is severe, particularly for those patients with spinal cord damage. Clinically, no long term therapeutic treatments have been developed that might inhibit scarring and promote neuronal growth. Consequently, the prevalence of patients permanently disabled from head and spinal cord injury is high, estimated at more than 1:1000 of the population of North America (Office of Technology Assessment USA, 1990). Ann Logan and Martin Berry define the mechanisms that underlie the wound healing response in the CNS and discuss the rationale for the development of novel therapeutic strategies. PMID- 8249156 TI - [The clinical and cytogenetic characteristics of the children born to persons with a history of 1st- and 2d-degree acute radiation sickness as a result of the accident at the Chernobyl Atomic Electric Power Station]. AB - We have examined 15 children (born in 1987-1988), whose fathers liquidated the aftereffects of the accident at Chernobyl Nuclear Power Plant and suffered from acute radiation sickness of the 1st and 2nd stages and 50 children of the control group. The obtained data showed that the number of small developmental abnormalities (stigmas of dysembryogenesis) increased as well as the chromatid aberration frequency as compared with the control group. PMID- 8249157 TI - [The cytogenetic disorders in the people who took part in the cleanup of the sequelae at the Chernobyl Atomic Electric Power Station and who lived continuously in areas with an unfavorable ecological situation]. AB - Cytogenetic disturbances are manifested now in persons who liquidated aftereffects of the Chernobyl disaster. The character and course of the disturbances depend on the ecological conditions of the region where liquidators live at present. Examination of a group of liquidators with nerve psychopathologic disorders and accompanying somatic pathology has revealed the presence of cytogenetic disturbances, mainly of the chromosome aberration type. At the same time, in persons who live under conditions of high environment pollution with ejections of industrial enterprises, the number of chromatid aberrations increases, which may be a result of action of chemical mutagens. A tight correlation is revealed between the level of cytogenetic disorders in lymphocytes and expressivity of the secondary immunodeficiency. Elimination of lymphocytes with unstable chromosome aberration is delayed when ecological pollution of the biosphere reaches the high level. PMID- 8249158 TI - [Limited genetic monitoring in Kiev in relation to the accident at the Chernobyl Atomic Electric Power Station]. AB - Frequency of innate developmental defects, perinatal death rate, spontaneous and induced abortions were statistically studied on the basis of primary medical information from archives of two largest maternity hospitals of Kiev for the period between 1969 and 1989. Significant variations in the chosen characters in different years were found, unidirectional changes in time being absent. No increase in the number of innate defects and frequency of spontaneous abortions after 1986 was found. It is shown necessary to introduce a system of unified registration of innate developmental defects adopted by an international register as well as to use more perfect mathematical methods for correct estimation of the processes occurring in human populations. PMID- 8249159 TI - [The protection of the gene pool of the population of Ukraine]. PMID- 8249160 TI - [Teratogenic effects in a population based on a model of the bone system of the human embryo]. AB - The dynamics of teratogenic effects was studied in population of town of Chernovtsy (Ukraine) on the model of the human embryo bone system. Visual and morphometric examinations of native intact bone preparations (including the ossification zones) of artificially aborted normal human embryos of 6-12 gestational weeks were made by the original method suggested by the author. The first stage of the study was performed in 1988 during acute total alopecia of children. The second stage was three and four years after the event. The obtained results have supported the evidence for a comparative rise in rate of chemically induced embryonal teratogenic defects in population of Chernovtsy. PMID- 8249161 TI - [The detection and assessment of the total mutagenic action of the aerosol portion of the chemical atmospheric contaminants from industrially developed cities of Ukraine]. AB - The studies on assessment of total mutagenic activity of chemical pollutants of atmospheric air in some Ukrainian towns with the mainly developed metallurgical industry were carried out using the Ames test with metabolic activation. It was established that the air samples taken in five towns during 12 months had mutagenicity to certain extent, both with and without metabolic activation. The degree of mutagenic effects' pronouncement among the towns tested was distributed as follows: Mariupol' > Zaporozhye > Donetsk > Krivoi Rog > Makeyevka. PMID- 8249162 TI - [The mutagenic effect of plasmid DNA pUC19 and its derivatives]. AB - For a better understanding of the mechanism of foreign DNA mutagenesis, it is essential to study the validity of the changing DNA structure for manifestation of its mutagenic action on Drosophila melanogaster (method Basc). The testing of three different kinds of plasmid DNA has been carried out. The injections of only plasmid DNA pUC18 lead to a significant increase in the X-linked recessive lethal frequency. It is important to check mutagenicity of every new recombinant DNA. PMID- 8249163 TI - [The mutagenic effect of the recombinant plasmid pBR322ins carrying a native and a modified human insulin gene]. AB - The mutagenic activity of pBR322 bacterial plasmid DNA and of pBR322ins and pBR322insN recombinant plasmid DNAs has been investigated in Blld-ii-FAF 28 line cultivated fibroblasts of Chinese hamster. The pBR322 bacterial plasmid DNA is shown to induce no resistant mutations to 6-mercaptopurine; the pBR322insN (the frameshift mutation in human insulin gene; this gene is supposed to have no expression in the cells because of frameshift mutation) induces neither such mutations as well. The pBR322ins recombinant plasmid carrying the native human insulin gene induces the gene mutations in this system. The influence of the insulin gene structure on the gene mutations induction is taken into account. The results obtained and the data of other authors illustrate a great interest of recombinant DNA molecules carrying transgenes in the field of mutation research. PMID- 8249164 TI - [Plasmid pSVc-myc-1 induces gene mutations and chromosome aberrations in cultured Chinese hamster cells]. AB - The induction of mutations to 6-thioguanine-resistance and chromosome aberrations by the plasmid pSVc-myc-1, carrying the activated cellular oncogene c-myc, isolated from a mouse plasmocytoma was studied in a cultured Chinese hamster cell line. The yield of HPRT- mutants and chromosome aberrations increased 1.6 times on the average after pSVc-myc-1 treatment. The mutagenic activity of pSVc-myc-1 was statistically significant. The role of mutagenic effects of activated cellular oncogenes in malignant transformations is discussed. PMID- 8249165 TI - [The characteristics of the chromosomal apparatus and functional state of the thyroid in amenorrhea in women]. AB - Girls-teenagers and women with menstrual disturbances were examined cytogenetically and clinically with special respect to the thyroid function. The rate of chromosomal pathology in the studied group was 29.5%. The rate of aberrant cells proved to be elevated in cases of primary and secondary amenorrhea as compared with the control. Chromatid aberrations were typical of the Turner syndrome (8.1% as against 2.1% in controls). In cases of menstrual disturbances the C-segment macrovariants were detected in chromosomes 9 and 16, so as inverted C-segments. The thyroid hyperplasia of the 1st, 2nd stages was detected at the rate of 81.2% among the teenaged girls with menstrual disturbances and TSH hypophysis-hyperfunction was typical of women with primary or secondary amenorrhea, providing evidence for the secondary origin of thyroid hormonal disturbances. PMID- 8249166 TI - [Chromosomal syndromes in the autosomal system and the problems of differential diagnosis]. AB - The karyotype of 2715 patients suspected of chromosome pathology has been studied. Chromosomal aberrations in the system of autosomes has been revealed. A diagnostic programme promoting a selective screening of chromosome pathology in nonspecialized medical institutions is developed. Medical-social rehabilitation programme for patients with chromosome syndrome is worked out. PMID- 8249167 TI - [Polymorphism of the C-band segments of chromosomes 1, 9 and 16 in the peripheral blood lymphocytes of patients with endometrial cancer]. AB - The paper presents results of investigation of constitutive heterochromatin of chromosomes 1, 9 and 16 in 23 patients with endometrial cancer and 5 women of the control group. The analysis was carried out on slides obtained by the routine method of the peripheral blood cell culture. The C-band segment variability was studied by the C-binding method. The investigation established an increase of polymorphism of constitutive heterochromatin in cancer patients, extreme variants and heteromorphism of homologues of chromosomes 1, 9 and 16. It is shown that polymorphism of the C-band on chromosomes 1, 9 and 16 in patients with cancer in their families increases as compared to the patients without cancer pathology in pedigrees. PMID- 8249168 TI - [An analysis of mutations in the 7th, 10th and 11th exons and of the polymorphism of the 4 nucleotide tandem repeats from the 3' end of the 6th intron of the CFTR gene in families from Ukraine with a high risk of mucoviscidosis]. AB - Mutations in the CFTR gene have been screened in 110 families from Ukraine with high risk of cystic fibrosis. Deletion F508 was found in 121 (55%) of 220 CF alleles. Among the rest mutant alleles (with the absence of delta F508) six other mutations occurring in the 7th, 10th, 11th exons of the CFTR gene were screened. In this way, 5 CF alleles may be characterized: 1 allele with R334W (the 7th exon) 1 with 1677 delTa (the 10th exon), 1 allele with G551D and 2 alleles with R553X (the 11th exon). We have screened 865 healthy donors for F508 from different regions of Ukraine. The frequency of heterozygous carriers in different regions ranged from 1:28 to 1:170. Strong linkage imbalance was found between polymorphic markers of 4 bp tandem repeats from the 3'-end of the sixth intron in the CFTR gene and deletion F508. PMID- 8249169 TI - [The mechanisms of the hereditary predisposition to ischemic heart disease]. AB - The hereditary predisposition of people to ischemic cardiac disease may be realized both by means of the lipid blood spectrum and through the break-down thrombosis processes. The argument mentioned is confirmed by the strong fixed correlation between patients who had infarction-myocarditis at young age and their children. PMID- 8249170 TI - [An evaluation of the mutagenic background and mutational variability of the population in a region with a high intensity of pesticide use]. AB - The mutagenicity of soils (on A. cepa and Tr. poludosa) and chromosome aberration level in population have been studied in two Uzbekistan areas. The obtained results are evidence of global contamination of the whole region by mutagenic and toxic chemicals. The mutation level in test objects and frequency of chromosome aberrations in peripheral blood lymphocytes in the examined persons have not been correlated. The condition of the area studied is classified by mutagenic contamination as "critical". PMID- 8249171 TI - [Dimatif modification of the cytogenetic effect in the peripheral blood lymphocytes of children subjected to a constant low-intensity radiation exposure]. AB - Primary results of cytogenetic monitoring of children living in regions of Ukraine contaminated after Chernobyl accident permit suggesting existence of radiation-induced modification of sensitivity of somatic cell chromosomes to mutagens. It may be supposed that interindividual and group difference in sensitivity of in vivo irradiated cells to additional induction of in vitro mutagen effect is explained not only by inherited properties, but also by preliminary irradiation. PMID- 8249173 TI - [The role of the genetic component in oncology]. AB - The paper presents information on fundamentals of oncogenetics, role of the clinical and genealogical method in oncology, in epidemiological genetics in particular. It is shown necessary to create registers of family cases of tumours proceeding from genetical data: risk groups on appearance of malignization. The ways for the further study of the role of a hereditary factors in tumour growth are outlined. PMID- 8249172 TI - [The immediate and late results of radiation effects on the occurrence of congenital defects in the locomotor apparatus based on screening data on newborns in the maternity hospitals of the city of Kiev]. AB - On the basis of screening of 15939 newborns at the maternity homes of Kiev after the accident at the Chernobyl atomic station it was found that the number of children with congenital malformation of the locomotor apparatus in 1991-1992 increased by 28.7% as compared with the data of the first postaccident years (1986-1987). Congenital pathology of the hip joint (dysplasia in average 23.5 per 1000 births, hip dislocation 3.6:1000) and foot deformity (7.9:1000) had the highest incidence with a tendency to increase at the latest years. PMID- 8249174 TI - Genetic events responsible for colorectal tumorigenesis: achievements and challenges. AB - Colorectal carcinogenesis is a multistep process that is accompanied by accumulation of changes in proto-oncogenes and tumor-suppressor genes. APC/MCC, RAS, DCC, p53 mutations and/or allelic losses, hyperexpression of c-MYC and RB genes, as well as other genomic alterations appear at characteristic stages of tumor development and are observed in most neoplasms. However, consideration of each of these abnormalities leaves many unanswered questions. The striking data on recurrent amplification of the RB tumor-suppressor gene as well as suppressive activities of protein kinase C and activated RAS genes, at least in some colon carcinoma cell lines, suggest the unusual effects of some signalling pathways in colonic epithelial cells. The results obtained to date indicate that distinct sets of genetic changes may underlie the development of colorectal tumors. PMID- 8249175 TI - Childhood leukemia in south-west Sardinia (Italy). AB - AIM AND BACKGROUND: Public concern on an increased incidence of childhood leukemia in SW Sardinia prompted the authors to an epidemiological investigation. METHODS: Incident childhood neoplasms observed in the Cagliari province (Sardinia -Italy) in 1974-1989 were registered. Expected cases of the most frequent childhood cancers were calculated for each town, based on the sex-and age specific incidence rates in the province. RESULTS: An excess risk of childhood acute lymphoblastic leukemia (ALL) was observed in Carbonia, a town located in the SW part of the province. The risk was highest in 1983-85, when 7 cases occurred versus 0.8 expected (RR = 8.7; 95% C.I. = 4.6, 16.3). No spatial clustering of ALL cases was observed within the town. CONCLUSIONS: A significantly higher than expected incidence of childhood ALL was observed in the town of Carbonia in 1983-85. In alternative to chance, possible exposure to environmental pollutants from a near industrial settlement is discussed as the cause of the observed excess, but it is far to be proven. Other hypotheses, including a viral infection in a population with increased susceptibility, as suggested for new urban settlements, cannot be discarded. PMID- 8249176 TI - Immunoendocrine therapy with interleukin-2 (IL-2) and medroxyprogesterone acetate (MPA): a randomized study with or without MPA in metastatic renal cancer patients during IL-2 maintenance treatment after response or stable disease to IL-2 subcutaneous therapy. AB - AIMS AND BACKGROUND: It is known that interleukin-2 (IL-2) activated cytotoxic lymphocytes require a cell-cell contact to exert their anticancer action. Therefore, the pronounced fibrosis that generally characterizes the neoplastic mass could counteract the action of cytotoxic lymphocytes. Some preliminary studies have shown that progesterone and its analogs may inhibit fibroblast proliferation. On the basis of such evidence, we have designed a clinical study with or without the progestational agent medroxyprogesterone acetate (MPA) in metastatic renal cancer patients in maintenance therapy with IL-2 following response or stable disease (SD) after two cycles of IL-2 subcutaneous immunotherapy, in an attempt to evaluate the influence of MPA on free-from progression (FPP) period. METHODS: The study included 30 consecutive patients who were randomized to receive IL-2 alone (3 million IU twice/day for 5 days/month subcutaneously) or IL-2 plus low-dose MPA (500 mg orally one day/week) without interruption until disease progression. RESULTS: A FPP period longer than 1 year was obtained in 8/14 patients treated with IL-2 plus MPA and in only 3/16 patients treated with IL-2 alone. The difference was statistically significant. On the contrary, no significant difference was seen in the mean number of lymphocytes and eosinophils, which was evaluated monthly. Finally, no hyperglycemic or thromboembolic complications occurred in patients concomitantly treated with MPA. CONCLUSIONS: This preliminary study would suggest that the concomitant administration of low-dose MPA may prolong the FFP period in metastatic renal cancer patients under maintenance therapy with IL-2. A longer follow-up will be required to evaluate the influence of MPA on overall survival. PMID- 8249177 TI - A phase I-II study of oral doxifluridine plus radiotherapy in radiosensitive tumors of the pelvic region. AB - AIMS AND BACKGROUND: Fluoropyrimidines have shown synergic effects in combination with radiotherapy in several tumor types. Doxifluridine is a novel 5-fluorouracil (5-FU) prodrug which is transformed into 5-FU in neoplastic tissue. This would imply enhancement of radiotherapy by 5-FU in neoplastic tissue, where the drug is concentrated higher than in surrounding healthy tissues. METHODS: A phase I-II study was carried out on 10 patients with radiosensitive tumors of the pelvic area (4 uterine carcinomas). Escalating doses of oral doxifluridine were administered in combination with standard radiotherapy to assess the efficacy and toxicity profile of the combined treatment. The 9 evaluable patients (3 groups of 3 patients each) received oral doxifluridine, at daily doses of 500, 750, or 1000 mg, for 6 consecutive weeks in combination with a standard (1.8-2.0 Gy) dose of radiotherapy. Assessment of physical and laboratory parameters was made at baseline, then weekly up to the end of the treatment and at follow-up. RESULTS: At the final evaluation, one patient with a diagnosis of uterine carcinoma showed a complete response that lasted 10 weeks. One patient had a partial response, and 7 patients had no change. The most frequent adverse events were gastrointestinal: 27 episodes of mild to moderate nausea/vomiting and diarrhea. Three patients complained of severe diarrhea of 5-7 days duration: all patients spontaneously recovered. There were no significant changes in laboratory or clinical parameters, and no serious toxicity requiring reduction or interruption of the radiotherapy. CONCLUSIONS: The maximum tolerated dose of oral doxifluridine in combination with standard radiotherpay was assessed at 1000 mg/patient/day (equivalent to 36-38 g monthly, previously reported as mTD in phase I studies). PMID- 8249178 TI - Chemotherapy with mitomycin C and vinblastine in pretreated metastatic breast cancer. AB - AIMS: In February 1986 we began a study to test the activity of mitomycin C (12 mg/m2) plus vinblastine (6 mg/m2) on day 1 of a 28-day cycle (MV) as second or third-line chemotherapy for metastatic breast cancer patients. METHODS: As of February 1988 the study was stopped after 26 patients had been enrolled. The median age of the patients was 54 years (range 35-78); all patients were progressive from chemotherapy; 15 (57.7%) patients were treated as second and 11 (42.3%) as third line; 19 (73.1%) patients had received anthracyclines as first (13 patients) or second-line (6 patients) chemotherapy; 18 (69.2%) patients had visceral involvement; 7 (26.9%) had one metastatic site, 11 (42.3%) two sites, 6 (23.1%) three sites and 2 (7.7%) four sites. RESULTS: Overall, 86 cycles were administered, with a median number of 3 cycles per patient. Toxicity was mild; hematologic side effects required discontinuation of treatment in 3 cases. Vomiting occurred in 3 (11.5%) patients, nausea in 5 (19.2%). Moderate neurologic toxicity was recorded in 6 (23%) patients. No complete and 3 partial responses were observed. The objective response rate was 11.5% (exact 95% confidence interval, 2.4-30.1). Responses occurred independently of disease-free interval, dominant metastatic site, response to previous chemotherapy, previous anthracycline and line of treatment; all responses were recorded in patients under 50 years of age. Kaplan-Meier estimated median time to progression and overall survival were 13 and 40 weeks, respectively. CONCLUSION: The MV regimen was well tolerated but showed little activity in pretreated metastatic breast cancer. PMID- 8249179 TI - Evaluation of the esthetic results of conservative treatment of breast cancer. AB - AIMS AND BACKGROUND: This paper describes the evaluation of a personal series of 106 breast cancer patients, aged 28-75 years (average, 54), treated between 1988 and 1971, to determine the success of a conservative protocol proposed as a means of combining radicality with a better cosmetic result, less alteration of the body image and an improved quality of life. METHODS: All the patients were operated with quadrantectomy and excision of axillary lymph nodes, followed by radiotherapy: 50 Gy to the whole breast plus 10 Gy on the tumor bed (boost irradiation). Adjuvant hormone and/or chemotherapy was also administered in 71/106 cases. The mean follow-up was 22.3 months. Cosmetic assessment was subjective and objective (asymmetry, loss of volume, retraction). All patients were photographed. RESULTS: The subjective judgements were: excellent 79, satisfactory 25, and poor 2, compared with objective assessments of 72, 20 and 14, respectively. Patients with poor esthetic results were almost all either T2 or treated with chemotherapy. CONCLUSIONS: Conservative treatment of breast cancer using integrated irradiation and surgery can therefore attain the required objective of eradication of the cancer while maintaining a satisfactory esthetic and therefore good psychologic outcome. PMID- 8249180 TI - Color-Doppler ultrasound in ovarian masses: anatomo-pathologic correlation. AB - BACKGROUND AND AIMS: In the radiologic assessment of ovarian masses, the major difficulty consists in the late recognition and lack of parameters for a differential diagnosis between benign and malignant lesions, especially in the post-menopause when the incidence of cancer is higher. The use of a transvaginal probe and the color-Doppler examination have recently improved the study of the female pelvis. This study is aimed to verify the possibility of the color-Doppler imaging to differentiate between malignant and benign ovarian lesions during transvaginal echographies, on the basis of the qualitative and quantitative characteristics of the vascular pattern of the ovarian lesions. RESULTS: Twenty six expansive ovarian lesions were studied: 8/26 showed no vascular signals and were considered benign as confirmed at histology. In the remaining lesions with some vascularization, the resistance index (RI) was evaluated: those with RI > 0.40 were considered benign, those with RI < 0.40 malignant. In 8/9 benign lesions and 7/9 malignant neoplasms, the results of color-Doppler were coherent with histology. The results showed a sensibility of 87.5% and a specificity of 88.8% for the transvaginal examination. CONCLUSIONS: The main advantages of the color-Doppler transvaginal examination are: the high frequency of visualization of the ovaries, even in postmenopausal patients; the definition of small lesions; the visualization of small parenchymal vessels, both physiologic and pathologic, and their quantitative analysis. The importance of the RI cutoff was critical for the differential diagnosis between benign and malignant lesions; we think that a cutoff of 0.50, instead of 0.40 proposed by other authors, would be far more appropriate. PMID- 8249181 TI - Hyperthermic potentiation of cisplatin cytotoxicity on solid Ehrlich carcinoma. AB - BACKGROUND: Hyperthermia produces marked effects on many biochemical parameters of tumor cells and has been reported to potentiate the effect of many drugs. We therefore evaluated the possible synergistic effect between hyperthermia and cisplatin against solid Ehrlich carcinoma. The study was based on the measurement of some biologic characteristics in tumor tissues, namely: DNA, RNA, and protein content and their rate of synthesis as parameters for nuclear damage; total lipids and cholesterol as parameters for membrane damage; acid-phosphatase and acid-ribonuclease as parameters for lysosomal damage; and tumor volume as a direct parameter for tumor growth. METHODS: Treatment of solid Ehrlich carcinoma by hyperthermia at 43 degrees C for 30 min for 3 successive days produced a 41.5% decrease in tumor volume, as well as a significant decrease in nucleic acids, protein contents and their rate of synthesis, in total lipids and cholesterol, and in acid-phosphatase and acid-ribonuclease. Chemotherapeutic management of the tumor by 5 mg/kg x 3 of cisplatin alone showed a continuous increase in tumor volume but at a lower rate than that of the untreated control. However, when cisplatin was given 1 h prior to hyperthermia, the tumor volume was significantly decreased by 82.6%. RESULTS: The effects observed on all the investigated parameter were intensified when cisplatin was combined with hyperthermia. The results obtained suggest that hyperthermia may enhance the penetration of cisplatin to its target site inside the tumor cells due to a membrane-damaging effect. The enhanced lethality of cisplatin on tumor cells may also be due to the inhibition of DNA repair processes by hyperthermia. PMID- 8249182 TI - Multiple primary tumors and familial cancers. A case report. AB - We herein report the case of a patient who had 4 primary tumors, 3 of which were malignant. His family had a high rate (26%) of cancer. The possible genetic factors that con produce these events are briefly discussed. PMID- 8249183 TI - Unusual breast edema and erythema during radiotherapy in the conservative approach of breast cancer. A case report. AB - The appearance of breast edema in the conservative approach of breast cancer is correlated with axillary dissection, and it is worsened by radiotherapy. In rare cases there are serious edema and erythema of the breast at the beginning of radiotherapy. We present a patient with edema and erythema of the breast with an unusual evolution after conservative surgical treatment of the breast cancer. Possible etiologies and published data are reviewed. PMID- 8249184 TI - Renal cancer after busulphan treatment for chronic myeloid leukemia. A case report. AB - The incidence of secondary malignancies following chemotherapy is progressively increasing, mostly due to prolonged survival of patients treated for primary cancer. So far, only 3 cases of solid cancer following busulphan administration have been reported. We describe the case of a patient who developed a renal cancer after 4 years of busulphan treatment for chronic myeloid leukemia. Immunosuppression rather than mutagenesis seems to be responsible for the emergence of second solid cancers in patients receiving busulphan. PMID- 8249185 TI - Spontaneous perforation of the rectum by carcinoma of the prostate. Case report. AB - A case of spontaneous perforation of the rectum by a locally advanced prostatic carcinoma is described. The mechanisms of rectal involvement by prostatic carcinoma are discussed, focusing attention on the difficulties in differential diagnosis with primary carcinoma of the rectum. PMID- 8249186 TI - Extraskeletal myxoid chondrosarcoma. A clinicopathologic and immunohistochemical study of two cases. AB - Two extraskeletal myxoid chondrosarcomas arising on the left lower extremity of two women, aged 48 and 72 years, respectively, are reported. One case mimicked clinically an organized hematoma. By light microscopy, they showed a lobular architecture. Anastomosing cords of cells lying in a myxoid background were the histologic hallmark. Based on conventional criteria, they were classified as neoplasms of low (G1) and intermediate (G2) grade malignancy. Both cases were strongly positive for S-100 protein and vimentin. Follow-up confirmed the low aggressiveness of this entity. PMID- 8249187 TI - Small bowel metastasis from squamous cell carcinoma of the larynx. A case report. AB - Small bowel metastases from squamous cell carcinoma of the head and neck are relatively rare and in some cases asymptomatic. In the case herein reported, small bowel metastasis from a laryngeal carcinoma caused a life-threatening complication. The surgical approach was effective in saving the patient's life. Death occurred for neck tumor progression without abdominal signs of secondary lesions. A review of the previously reported cases is provided. PMID- 8249188 TI - Adenosquamous carcinoma of the colon. Case report of an unusual type. AB - Primary adenosquamous carcinoma of the colon is a rare entity. Its prevalent site of origin is at the level of the proximal portion of the large bowel. The tumor presents in young patients and follows a highly aggressive course. We present an extremely unusual case of adenosquamous carcinoma in which the classical mucinous and signet-ring cell adenocarcinoma was associated with the squamous component. The prognosis of this rare neoplasm is very poor. PMID- 8249189 TI - Plasma, erythrocyte and urinary selenium levels in sickle cell homozygotes and traits. AB - Plasma, erythrocyte and urinary selenium levels were determined in 21 sickle cell homozygote patients, 20 siblings with sickle cell traits, 29 parents and in 21 healthy controls with an HbAA pattern. The mean levels of plasma and erythrocyte selenium and the daily urinary selenium depletion were found to be lower in the HbSS patients than in the controls (p < 0.02; p < 0.001; p < 0.001, respectively). Urinary selenium depletion was found to be lower in the sickle cell trait siblings than in the controls (p < 0.001), but the plasma and erythrocyte selenium levels were close to normal values (p > 0.05). The mean erythrocyte selenium levels were found to be decreased in the parents as compared with the controls (p < 0.05), which indicates that urinary selenium losses may be replenished primarily by sources in the plasma and in the erythrocytes before stores in the other parts of the body are used. PMID- 8249190 TI - Pulmonary dysplasia with renal malformations: a common connective tissue disorder? AB - The effect of D-penicillamine (DPA) on the development of lung and kidney tissues was evaluated to determine whether the association of lung and kidney malformation results from a common generalized disorder of connective tissue in the fetal rat. Ten animals received a daily dose of 300 mg DPA in their drinking water during the last six days of their gestation. Ten other pregnant rats were used as controls. At the end of the gestation period all the animals were delivered by cesarean section. We found that DPA treatment during gestation caused marked growth retardation in the offspring. Although the lung weight to body weight ratios did not differ between the study and control groups, the left kidney weight to body weight ratios were found to be lower in the study group. Histologic sections showed pulmonary dysplasia with bronchomegaly, cystic alveoli, atelectasis, vascular aneurism, perivascular loose of connective tissue, small hemorrhagic foci in the lungs and caliechtasis. We hypothesize that a common connective tissue disorder induced by DPA during intrauterine life may cause lung and kidney malformations. PMID- 8249191 TI - Griscelli's syndrome: clinical features of three siblings. AB - Three siblings diagnosed as having Griscelli's syndrome (GS) are presented. The clinical features were partial albinism, silvery hair and absence of giant granules in the white blood cells. The diagnosis of GS was confirmed intra-vitam in the youngest sibling (propositis) at the age of nine months by the demonstration of irregular clumps of pigment in the hair shaft, and in particular melanocytes engorged with melanosomes in the skin biopsy, findings characteristic of this syndrome. A retrospective diagnosis of GS was made in the older two siblings. The first sibling died at the age of two, having a clinical picture suggestive of bulbar poliomyelitis. However, no tissue was available for histopathologic examination. The second sibling developed fever, jaundice, seizure, hepatosplenomegaly and lymphadenopathy and died at the age of six. Postmortem examination of this sibling revealed lymphohistiocytosis in the liver and spleen. The propositus died at the age of five following development of central nervous system involvement. Immunologic studies were not available in the first sibling. The IgG level was slightly low and the T-lymphocyte number was normal in the second sibling. The propositus had normal serum immunoglobulin levels and T-cell numbers and skin tests were positive with phytohemagglutinin and candida. PMID- 8249192 TI - Anesthetic management of a patient with hereditary fructose intolerance and phenylketonuria. AB - This is a report of a five-year-old girl with phenylketonuria (PKU) and hereditary fructose intolerance (HFI) who underwent elective strabismus surgery. PKU and HFI are two inborn errors of metabolism which have an autosomal recessive mode of inheritance. This case report describes the anesthetic features of a patient with PKU and HFI, each defect requiring specific anesthetic management. PMID- 8249193 TI - Blue rubber bleb nevus syndrome: report of a case. AB - A case of blue rubber bleb nevus syndrome (BRBNS) in a 17-year old boy with multiple cutaneous and gastrointestinal hemangiomas is presented. The patient had iron deficiency anemia and pica. In addition, we describe, for the first time, left renal artery, right hepatic artery, and ureteral abnormalities in a patient with BRBNS. PMID- 8249194 TI - Unusual appearance of the liver on ultrasonography and computed tomography in a patient with cystic fibrosis. AB - A seven-year-old boy with cystic fibrosis (CS) who presented with abdominal pain is reported. Ultrasonographic and computed tomographic studies of the upper abdomen revealed unusual liver findings. An ultrasound scan showed a liver that was exceedingly heterogeneous and a mixed echo pattern with dominant hyperechogenicity. Computed tomography showed large, multiple hypodense cyst-like lesions in the liver. Using the ultrasound scan as a guide, a needle biopsy was performed. The pathological findings were in accord with the findings obtained from ultrasonography and computed tomography, and were consistent with pathological findings seen in CS cases. PMID- 8249195 TI - Unusual neurologic complications of typhoid fever (aphasia, mononeuritis multiplex, and Guillain-Barre syndrome): a report of two cases. AB - Although typhoid fever can frequently lead to such neurological manifestations as confusion, delirium, and encephalopathy, Guillain-Barre syndrome, aphasia and mononeuritis multiplex occur very rarely. In this report, we describe two patients with typhoid fever who developed these unusual complications. PMID- 8249196 TI - Subependymal giant cell astrocytomas in tuberous sclerosis. AB - Two cases of tuberous sclerosis with subependymal giant cell astrocytoma are presented. This rare autosomal dominant disorder was also detected in family members of the patients who had never had any symptoms of cerebral involvement. Both patients underwent surgery because of signs of increased intracranial pressure. PMID- 8249197 TI - Megalourethra associated with prune-belly syndrome. AB - A 14-day-old male infant with megalourethra is presented because of the rarity of the anomaly and its association with prune-belly syndrome. The lax, wrinkled appearance of the abdomen, bilateral cryptorchidism and severe dilatation of the urinary system are features included in the classic triad of the prune-belly syndrome. Our patient had the scaphoid variety of megalourethra since the penis appeared elongated and floppy in the fusiform form. PMID- 8249198 TI - Evoked potentials in Guillain-Barre syndrome. AB - A prospective evaluation was made in a total of 19 patients with Guillain-Barre syndrome (GBS) by neurologic examination, visual evoked potentials (VEP), brain stem auditory evoked potentials (BAEP) and somatosensory evoked potentials (SEP) during the acute phase of the disease, three weeks later, and then again three months after the onset of symptoms. The results of electrophysiologic studies of the patients were compared with those of 19 healthy children. There were no statistically significant differences found in the mean VEP and BAEP values between the patient and control groups. However, the VEP and BAEP values were abnormal during the first and second stages of the disease in only a few patients. There was a statistically significant prolongation of the response in tibial and median SEP during the first and second stages of the disease when compared with the third stage and the control group. The Erb-Cervical 7 (C7) interwave latency was elevated in the median SEP. It was concluded that the prolongation of the response in SEP was diagnostically important in the acute phase of GBS. There was no correlation found between SEP and clinical progression. PMID- 8249199 TI - Comparative therapeutic results of penicillin plus chloramphenicol versus ampicillin plus sulbactam in childhood meningococcemia. AB - Fifty-seven cases of meningococcemia were evaluated retrospectively. The age of the patients ranged between 2 and 17 years. Of the 57 patients investigated for the efficacy of antibiotic treatment, 31 (54.4%) were treated with benzylpenicillin plus chloramphenicol and 26 (45.6%) with ampicillin plus sulbactam. Patients with criteria for a poor prognosis (presence of disseminated intravascular coagulation, low arterial blood pressure, and altered consciousness) were divided equally into two treatment groups. There were no statistically significant differences between the two treatment groups except for the higher incidence of convulsion in the group given penicillin plus chloramphenicol. The mortality rate was 19.3 percent for patients treated with benzylpenicillin plus chloramphenicol and 7.6 percent for patients treated with ampicillin plus sulbactam (p = 0.19; overall mortality rate 14%). PMID- 8249200 TI - Long-term results of patients with congenital complete atrioventricular block. AB - Congenital complete atrioventricular block is an uncommon disorder with a prognosis which is usually favorable. The disorder is sometimes associated with syncope, sudden death or cardiac pacing. It is difficult to determine the patients at risk of sudden death. In this retrospective study, hospital records of children who had been admitted to the Hacettepe University Children's Hospital between 1970-1990 for evaluation of complete heart block were examined. The study population, consisting of 39 children, 27 males and 12 females, had electrocardiograms consistent with complete heart block. These patients, diagnosed as having congenital complete A-V block, had an otherwise normally structured heart, and were followed up for a period of from one month to 15 years. Age at diagnosis ranged from 27 days to 17 years (mean: 86.9 +/- 48 months). Of the 14 patients with symptoms (five with syncope, eight with exercise intolerance and one with presyncope), nine were paced electively and have done well. Clinical and laboratory features of the asymptomatic and symptomatic groups were compared to evaluate potential risk factors. PMID- 8249201 TI - Oral rehydration of infants with hypernatremic dehydration due to acute gastroenteritis. AB - Twenty-five infants with hypernatremic dehydration due to acute gastroenteritis were given oral rehydration therapy (ORT). The patients received a glucose electrolyte solution (such as that recommended by the World Health Organization) over six hours (2:1 rotating method). Twenty-three patients were successfully rehydrated within 48 hours after onset of therapy, while the two remaining patients attained normal serum Na+ levels within 72 hours. Acidosis was noted in 10 patients which disappeared in 24 hours. PMID- 8249202 TI - [Treatment of asthma with theophyllines]. PMID- 8249203 TI - [Doppler evaluation of Saint Jude aortic prostheses (pressure gradient and functional surface)]. PMID- 8249204 TI - [Evaluation of the sensitivity of routine obstetrical echography in the prenatal diagnosis of major fetal anomalies]. PMID- 8249205 TI - [Characteristics of pregnancy and delivery in the cardiac patient]. PMID- 8249206 TI - [Sarcoidosis in a dermatology service]. PMID- 8249207 TI - [Hematological characteristics in primary myelodysplastic syndromes: retrospective study of 14 cases]. PMID- 8249208 TI - [Granulomatous gastritis. 8 case reports]. PMID- 8249209 TI - [Bilateral spontaneous rupture of the extensor apparatus of the knee in chronic renal insufficiency]. PMID- 8249211 TI - [New strategies of the Medical Research Council of Canada]. PMID- 8249210 TI - [Hirsutism and pregnancy. A case report]. PMID- 8249212 TI - [In cardiology: no miracles, but regression of mortality with therapeutic options that multiply]. PMID- 8249213 TI - [Hyperthyroidism in a young, female patient]. PMID- 8249214 TI - [Arms control: a question of public health]. PMID- 8249215 TI - [Peer review: the more it changes...]. PMID- 8249216 TI - [Periodic medical examination, update 1992. 3. Anti-HIV antibody screening. Canadian work group on the periodic medical examination]. PMID- 8249217 TI - [Perceptions of mental illness in Quebec residents]. AB - A transverse survey was conducted with 772 Quebecers in September 1992. Its purpose was to evaluate the level of awareness of the general public about mental illness, the general knowledge of these illnesses and their treatments and the attitudes towards mentally ill patients. Results show that half of the interviewed subjects still feel that they are not informed enough about mental illness and even well informed people have a poor general knowledge of mental illnesses and their treatments. Finally, more than 65% of the surveyed population show positive attitudes towards psychiatric patients, which seems to be an improvement in comparison with the past years. The author believes that physicians should give more information on mental illness and particularly on the role of drugs in the treatment of mentally ill patients. It seems also important to put more emphasis on information to low and middle class people. PMID- 8249218 TI - [Pregnant women's knowledge of prematurity in Quebec]. AB - The objective of this study was to determine pregnant women's knowledge level concerning premature birth. During July of 1990, 465 pregnant women completed a questionnaire evaluating their knowledge of the following dimensions of premature birth: risk factors, symptoms, epidemiology, prevention and self-assessment of risk. The subjects were all francophones, at 16 weeks of less of pregnancy, with characteristics comparable to all pregnant women in Quebec. The study participants considered premature birth to be a serious health problem, and over estimated the premature birth rate. Fifty-seven percent thought that the rate was 17% or greater. Eighty percent associated premature birth with some morbidity and mortality risks. On the other hand, the risk factors concerning premature birth were poorly known; approximately 50% of the women did not recognize twin pregnancy and a past history of premature delivery as risk factors. Moreover, a third of the women did not recognize the signs and symptoms of premature rupture of membranes and of premature labor. In conclusion, a high proportion of pregnant women in Quebec do not have even minimally adequate knowledge levels concerning the prevention or risks of premature birth. PMID- 8249219 TI - [Food poisoning caused by a chemical product]. PMID- 8249220 TI - Evaluation of upper urinary tract dynamics by diuresis renography in patients receiving urinary reservoir operation. Comparison of full and empty reservoir. AB - Upper urinary tract dynamics was evaluated using diuresis renography during full and empty reservoir in 14 patients (11 men, 3 women, 13-70 years old) with intestinal urinary reservoir or intestinal bladder augmentation. Types of operation were Mainz pouch urinary diversion 5, Mainz neobladder to urethra, 5, Kock pouch urinary diversion 2, and Mainz bladder augmentation 2. Diuresis renography was performed using 99mTc-DTPA and furosemide during full and empty reservoir, and several parameters (Tmax, T75, T50, GFR) in addition to the patterns of renogram were evaluated. In the normal control, none of the parameters of the diuresis renogram with full bladder differed from those with empty bladder. In 5 of 14 patients, abnormal waves on cystometry (CMG) were found, and in 4 of these 5 patients, the patterns of diuresis renogram of full reservoir were worse than those of empty reservoir. However, the patterns of diuresis renogram of full reservoir were not different from those of empty reservoir in 9 patients without abnormal waves on CMG. In conclusion, renal injury may easily occur in patients with intestinal reservoir and abnormal waves on CMG. PMID- 8249221 TI - Vesicourethral function after surgery for uterine cancer: predictive value of postoperative maximum urethral closure pressure on residual urine. AB - Using a urodynamic technique, vesicourethral function was evaluated in 30 patients who had undergone surgery for uterine cancer. In all 28 patients treated with radical hysterectomy, detrusor function was damaged irrespective of the use of the Cavitron ultrasonic surgical aspirator (CUSA), whereas, in 2 patients treated with semiradical hysterectomy, the function was preserved. Vesicourethral functions 3 weeks after the operation in all 28 patients treated with radical hysterectomy showed underactive detrusor function with low compliance. In 10 out of these 28 patients, postoperative maximum urethral closure pressure (MUCP) did not decrease, without relation to the use of CUSA nor to the presence of detrusor sphincter dyssynergia, and more than 5 weeks were required to reduce residual urine to less than 50 ml. On the other hand, in the remaining 18 patients, postoperative MUCP decreased, and residual urine decreased to less than 50 ml by 5 weeks after the operation. From the results obtained here, we can conclude that the evaluation of MUCP at an early phase after the operation is a useful tool to predict complications caused by residual urine, such as urinary tract infection, irreversible alteration of detrusor function or deterioration in renal function. PMID- 8249222 TI - Antimicrobial treatment for chronic prostatitis as a means of defining the role of Ureaplasma urealyticum. AB - A study was conducted to assess the clinical and microbiological effects of antimicrobial treatment for chronic prostatitis as a means of defining the role of Ureaplasma urealyticum. Significant U. urealyticum cells were considered to be isolated from the prostates of 18 of 143 prostatitis patients. These patients with ureaplasma-associated prostatitis were randomly treated with either ofloxacin or minocycline for 2 weeks; 4 patients were excluded due to voluntary withdrawal. U. urealyticum was eradicated in all the patients. Symptoms were resolved in 10 patients, and leukocytes in expressed prostatic secretion were cleared in 4 patients; both drug treatments revealed similar results. Even if we exclude 3 patients with significant coexistent Staphylococcus epidermidis cells before treatment, 3 of 11 patients evaluated showed complete resolution of symptoms and clearance of leukocytes in expressed prostatic fluid. These results suggest that U. urealyticum is a causative organism in some patients with chronic prostatitis. PMID- 8249223 TI - Mass screening for prostate cancer and the bias relating to survival rate. AB - Since 1981 we have been studying prostate cancer (Pca) by mass screening (MS) in the Gunma Prefecture, Japan. From 1981 to 1990, 9,067 subjects (total 15,451) were examined in connection with this project and 121 subjects were diagnosed as having Pca. The presence of Pca in 87 cases was confirmed at their initial MS (initial group); 34 cases were confirmed either the year after or several years after the first MS (repeat group). To evaluate the significance of MS for Pca, in this group of patients, the effects of 'lead time bias', 'self-selection bias' and 'length bias' on the survival rate were compared to Pca patients detected in the hospitals of the Gunma Prefecture during the same period. The survival curves of MS cases for each stage of the disease were better than those of the controls. Only in stage D was there a significant difference between the two groups. However, this MS curve decreased from the 4th year in the same manner as the control curve from the start. The clinical characteristics (age, pathological differentiation, prostatic acid phosphatase, gait disturbance, erythrocyte sedimentation rate, chronic disease, and pain in the patients with stage D disease) were compared between both groups. All characteristics studied, except age, in the MS group were clinically more favorable compared to those of the controls for each stage or in total. Moreover, the relative survival rate of subjects examined by MS was greater than 1.0. No significant difference was found in stage distribution between the initial group and the repeat group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249224 TI - Total blood and urine protein of urolithiasis patients with renal calculi of different chemical composition. AB - We determined total blood protein (TBP) and total urine protein (TUP) in healthy subjects and urolithiasis (UL) patients with renal calculi of different chemical composition: phosphate (CaP), oxalate (CaOx) and urate (HUr). We discussed the peculiarities of TBP and TUP distribution curves obtained and showed that the data on TBP and TUP do not make it possible to reliably single out patients with UL or determine the chemical composition of calculi in their kidneys. However, it was established that the comparison of TUP measurement results for UL patients using the Ponseau-S and sulfosalicylic acid methods makes it possible to reliably separate patients with Ca-containing calculi (CaP and CaOx) from those with HUr calculi. The explanation to this phenomenon using the data on the fractional composition of TUP and the organic matrix of those patients' calculi is given. PMID- 8249225 TI - Local chemolysis of obstructive uric acid stone with 0.1 M THAM and 0.02% chlorhexidine. AB - We used a modified chemolytic solution by combining 0.1 M THAM (tris[hydroxymethyl] aminomethane) with 0.02% chlorhexidine to treat 10 obstructive uric acid stone patients. A total of 23 local chemolysis sessions were performed. Five patients received 13 sessions (average: 2.6) as an adjuvant chemolysis after extracorporeal shock wave lithotripsy 5 patients received 10 sessions (average: 2) as a primary treatment. The average time of local chemolysis was 2.3 days, ranging from 1 to 4 days. In the 23 local chemolysis sessions, only one patient developed two episodes of fever (8.7%), which were managed successfully by conservative treatment. The mean values of effective renal plasma flow of treated kidney was increased from 108.2 to 155.6 ml/min (p < 0.05). The stone-free rate assessed by IVP and ultrasonography at 3 months follow up was 80%. Our results indicate that the combined use of 0.1 M THAM and 0.02% chlorhexidine is an effective and safe method to treat obstructive uric acid stones. PMID- 8249226 TI - Effects of shock waves on the male reproductive system. AB - Although many reports concerning the effects of shock waves on the human body have been published, their effects on the male reproductive system are not well known. This report examines the effects of shock waves on human spermatozoa and rat testes. Semen samples were placed in water at 37 degrees C and 100, 200, 500, and 1,000 shock waves were applied. In proportion to an increasing number of shock waves, sperm motility and the percentage of viable sperm decreased, while the percentage of short-tailed sperm increased. In a second study 500 or 1,000 shock waves were applied to the testes of infant and adult male Wistar rats. Hematomas were observed macroscopically in the testes immediately after shock wave treatment. Light microscopy showed rupture of the basement membrane of the seminiferous tubules and outflow of spermatogenic cells. After 5 weeks, the treated testes appeared atrophic, with the absence of spermatogenic cells in the seminiferous tubules, except for Sertoli cells. Our data indicate that spermatozoa and the testis appear to be structurally damaged by shock waves. PMID- 8249227 TI - Renal cell cancer and concomitant transitional cell cancer of the renal pelvis and ureter in the same kidney--report of 4 cases and review of the literature. AB - Three patients (1 man, 81 years old; 2 women, 56 and 61 years old) had renal cell cancer and transitional cell cancer in the same kidney, and another patient (female, 77 years old) had renal cell cancer and transitional cell cancer in the ipsilateral ureter. Simultaneous occurrence of renal cell cancer and transitional cell cancer is an extremely rare entity with only 15 cases having been reported. PMID- 8249228 TI - Papillary renal cell carcinoma associated with autosomal dominant polycystic kidney disease. AB - A case of papillary renal cell carcinoma in a patient suffering from autosomal dominant polycystic kidney disease is described. The occurrence of papillary renal cell carcinoma in polycystic kidneys in the literature is reviewed and the implications of operative management and further surveillance in case of a papillary renal cell carcinoma are discussed. PMID- 8249229 TI - Radical cystectomy for invasive squamous cell bladder cancer in a renal transplant patient. AB - A rare case of invasive squamous cell carcinoma of the urinary bladder developing 5 years after cadaveric renal transplant is described. The technical precautions to be taken while performing a radical cystectomy in such a situation is highlighted. This patient required chemotherapy in the post-operative period, and the role of chemotherapy in the presence of immunosuppressive therapy is discussed. PMID- 8249230 TI - Renal milk of calcium: review of the literature and report of 5 cases. AB - To our knowledge only a few cases of renal milk of calcium have been reported. We have encountered 5 such patients. The diagnosis depends upon the demonstration of half-moon-shaped calcification in the lateral decubitus or upright roentgenograms. This condition should be differentiated from a renal calculus to avoid unwarranted surgery or extracorporeal shock wave lithotripsy. PMID- 8249231 TI - Successful fibrinolytic therapy using tissue plasminogen activator in acute renal failure due to acute thrombosis of bilateral renal arteries. AB - A 46-year-old housewife with regurgitation of mitral valve developed acute renal failure due to acute thrombosis of bilateral renal arteries after cardiac catheterization. A regional fibrinolytic therapy using transarterial infusion of tissue plasminogen activator (t-PA) was performed at 8 h after the onset, and was successful. She could be taken off hemodialysis 2 weeks after the fibrinolytic therapy, and could be successfully operated on with mitral valve replacement 7 months after that. Three years after mitral valve replacement, her serum creatinine was 148 mumol/l. This is the longest follow-up case after regional transarterial fibrinolysis of acute thrombosis of renal artery using t-PA, and this therapy is a safe and good one for acute thrombosis of renal artery. PMID- 8249232 TI - Squamous cell carcinoma of the prostate. AB - A case of the rare squamous carcinoma of the prostate is herein presented. Squamous cancer accounts for 0.5-1% of prostatic malignancies. The patient died of systemic metastases 5 months after diagnosis. Review of the literature suggests that such a cancer is highly aggressive and responds poorly to any mode of therapy, perhaps because of a different cell of origin. PMID- 8249233 TI - Potbellied pigs. Miniature porcine pets. AB - The potbellied pig was introduced into the United States in 1985 and has since become a desirable pet throughout the country. When seeking medical or husbandry advice, pig owners often seek the counsel of their local small animal practitioner. The practitioner may feel at a loss to adequately respond to requests for veterinary care because they do not remember pig medicine from veterinary school. Large animal practitioners, who have knowledge of the medicine and surgery of commercial pigs, often do not like caring for potbellied pigs because they are pets and not production animals. This article is intended to provide the small animal practitioner with some practical knowledge in the husbandry and medical care of the pet pig, also known as the yuppy puppy. PMID- 8249234 TI - Exotic pet medicine I. Snakes. AB - Of the 6,000 extant species of reptiles, approximately 2500 are snakes. Several dozen species of snakes have become popular in the pet trade, and to meet the increased demand, more and more species are being bred in captivity. Along with this popularity comes the need for more sound veterinary expertise. Although much biomedical information on snakes exists in the literature and can be found in texts on reptile medicine, few reviews have tried to consolidate the literature on this subject. The purpose of this article is to bring together information on the most pertinent aspects of snake medicine and disease. PMID- 8249235 TI - The medical care of iguanas and other common pet lizards. AB - The majority of health problems in captive lizards result from improper diets, caging, and environmental conditions. This article discusses recommended husbandry and management techniques. A clinical approach to evaluating the health status of lizards, including assessing patient history, physical examination, clinical pathology, anesthesia, and surgery, is reviewed. Common health maladies of captive lizards are discussed, and rapid diagnosis and treatment are emphasized. PMID- 8249236 TI - Biology and medicine of turtles and tortoises. AB - Turtles and tortoises are unique reptiles that are gaining popularity as pets. Their anatomy and defense posture hinder, but do not preclude, clinical assessment and performance of routine diagnostic and therapeutic procedures by the clinician. A basic working knowledge of chelonian taxonomy, anatomy, physiology, husbandry, common diseases, and therapeutics will enable the veterinarian to provide health care to this order of reptiles. PMID- 8249237 TI - Exotic pet medicine I. Amphibians. AB - This article presents information on the basic taxonomy, biology, medicine, surgery, and anesthesia of amphibians. Included for quick reference are tables listing commonly kept amphibians, hormones for induction of reproduction, diseases, and clinical signs, commonly used pharmaceuticals and chemicals, nutritional diseases, and parasites. PMID- 8249238 TI - Diagnostic imaging of exotic pets. AB - Radiographic, ultrasonographic, and computed tomographic (CT) imaging are important diagnostic modalities in exotic pets. The use of appropriate radiographic equipment, film-screen combinations, and radiographic projections enhances the information obtained from radiographs. Both normal findings and common radiographic abnormalities are discussed. The use of ultrasonography and CT scanning for exotic small mammals and reptiles is described. PMID- 8249239 TI - Principles and techniques of anesthesia and analgesia for exotic practice. AB - Principles and techniques of anesthesia and analgesia for small animal exotic practice are described. The use of regimens based around inhalation anesthesia are recommended over injectable anesthetics. Techniques of perianesthetic monitoring and support are described and emphasized. The article concludes with a discussion of anesthesia for specific exotic animals likely to be encountered in small animal practice. PMID- 8249240 TI - Relationship between pharmacological properties and clinical efficacy of ruminant anthelmintics. AB - The purpose of this review article is to establish a relationship between pharmacological properties and clinical efficacy of the most widely used broad spectrum veterinary anthelmintics. The impact of drug pharmacokinetics/pharmacodynamics on anthelmintic efficacy and resistance is discussed. We review the clinical pharmacokinetics of the currently available anthelmintics used in livestock-imidazothiazoles, tetrahydropyrimidines, benzimidazoles/pro-benzimidazoles, salicylanilides (closantel) and avermectin type compounds. Understanding the pharmacokinetic and metabolic behaviour of broad-spectrum anthelmintics in the host, and factors modulating that behaviour, is highly important for maximizing anthelmintic utility and efficacy. PMID- 8249241 TI - A comparison of the antigen detection ELISA and parasite detection for diagnosis of Trypanosoma evansi infections in camels. AB - Two herds of 60 camels each, living in Trypanosoma evansi endemic areas, were selected and studied for a period of 18 months. Animals in one herd were treated prophylactically with quinapyramine prosalt (May and Baker, Dagenham, UK), while those in the other herd were treated individually with quinapyramine dimethylsulphate (May and Baker, Dagenham, UK) when proven parasitaemic. The herd on prophylaxis was sampled for antigen and patent infection monthly. The other herd was sampled weekly for patent infection and fortnightly for antigen. The results obtained could be divided into four categories. The first category comprised cases (52 out of 61) in which the presence of trypanosome antigens could be correlated with parasitological diagnosis. In 80% of these animals the antigens disappeared from the circulation within a period of 30 days following chemotherapy. The second category comprised those animals with parasitologically proven infections but which did not have antigens in their sera. This was observed in nine camels, seven of which were from the herd that was being examined weekly for the presence of trypanosomes. These were considered to be animals in early infection, as the subsequent sera were also negative for anti trypanosome antibodies and immune complexes. The third category comprised camels which were antigen-positive but aparasitaemic. Sera from these animals were also positive for anti-trypanosome antibodies, indicating that antigen-positivity was a true reflection of trypanosome infections in these animals. The last category comprised pre-weaned camel calves which appeared to have some form of protection against trypanosomiasis, as evidenced by the absence of trypanosomes, antigens and antibodies throughout the early period of their lives. Only occasional antigenaemia was found in a few calves. It is concluded that trypanosome antigen detection may give a more accurate idea of the prevalence of T. evansi infections than does whole parasite detection. PMID- 8249242 TI - Kinetic responses of parasite-specific antibody isotypes, blood leucocyte pattern and lymphocyte subsets in rats during primary infestation with Fasciola hepatica. AB - Antibody responses, blood leucocyte and splenic lymphocyte subset patterns were studied during a primary infection with Fasciola hepatica in the rat. The infection induced parasite-specific IgM by 2 weeks after infection. High levels of IgM antibodies were maintained for many weeks. The IgE response was biphasic with peaks at 5 and 9 weeks after infection which were correlated with different phases in the development of F. hepatica in the rat. Both IgG2a and IgG1 antibodies were detected but the titre of IgG2a augmented slightly and rose more slowly than did that of IgG1. There was a rise in neutrophil and eosinophil numbers. Neutrophils did not increase before the fourth week but eosinophil numbers were raised by the second week after infection and remained high during the whole migratory phase of the parasite. In the spleen, the percentage of B lymphocytes increased and there was a decrease in the percentages of CD4+ and CD(8+)-like T lymphocytes. These results suggested that, in the rat infested with F. hepatica, TH2-like lymphocytes could be preferentially stimulated, as has been reported in murine schistosomiasis. PMID- 8249243 TI - Oesophagostomum radiatum: the effect of different adjuvants on vaccination with a partially purified protective antigen. AB - The components of a soluble extract of adult Oesophagostomum radiatum were separated into four fractions by gel filtration chromatography on a Superose-12 column. The second fraction had a molecular weight range of approximately 60,000 65,000 as judged by SDS gel electrophoresis. Vaccination of calves with this fraction significantly (P < 0.05) reduced faecal egg output after challenge infection. In addition, the packed cell volume of vaccinated calves was higher than that of control calves (P < 0.05). The effect of three adjuvants on the vaccine activity was compared. Fraction 2 presented in dextran sulfate reduced the faecal egg count by 93.6% (P < 0.05), Fraction 2 in Freund's complete adjuvant reduced the faecal egg count by 27% (P = N.S.). Quil A was ineffective as an adjuvant. PMID- 8249244 TI - Sarcocystis-like organisms in musculature of a domestic dog (Canis familiaris) and wild dogs (Lycaon pictus) in Kenya. AB - Three of six wild dogs (Lycaon pictus) which died of rabies, and one Alsatian dog which died of adenocarcinoma, were found to harbour sarcocysts in their cardiac and/or skeletal muscles. The bradyzoites and metrocytes of the cysts were smaller than those seen in a lion infected with Microbesnoitia leoni and the common Sarcocystis spp. found in ruminants. It is proposed that Lycaon pictus is the natural intermediate host of the unidentified Sarcocystis-like species. PMID- 8249245 TI - The effect of prostaglandin inhibition on the development of pulmonary pathology associated with dead Dirofilaria immitis. AB - Flunixin meglumine was used to examine the effect of prostaglandin inhibition on the pathogenesis of Dirofilaria immitis in the pulmonary arteries of dogs. Immunopathological reactions to dead filariae were monitored by light and transmission electron microscopy and serology. Lung lesions in prostaglandin inhibited dogs exposed to dead filariae were enhanced compared to control dogs. This was associated with the persistence of parasitic antigen in lung tissue and in the blood. Serology demonstrated that after insertion of D. immitis in treated dogs, antibody levels did not change, while immune complex and antigen levels increased. These results indicate that prostaglandin may have a protective effect on the way the lung reacts to dead D. immitis, and that altered dynamics of the antigen processing may well contribute to the associated lung lesions. PMID- 8249246 TI - Effect of temperature and relative humidity on survival of eggs and infective larvae of Ostertagia circumcincta. AB - Survival of Ostertagia circumcincta eggs in faeces kept at 4 degrees C and of third stage larvae (L3) kept in distilled water at -50, -25, -10, 4, 16, 25 and 35 degrees C was studied. The effect of relative humidity (RH) of 95, 75, 50 and 30% at 16, 25 and 35 degrees C on the survival of L3 was also studied. The survival of eggs at 4 degrees C was high during the first week, followed by a gradual decline to 10% by Day 22 of storage. The L3 survived for 7 weeks at -50 degrees C, 9 weeks at -25 degrees C, 13 weeks at -10 degrees C, and 7 weeks at 35 degrees C. By 16 weeks, at the termination of the experiments, 74%, 76% and 23% of L3 were still alive at 4 degrees C, 16 degrees C and 25 degrees C, respectively. At all temperatures tested, the L3 survived longer at RH of 30 and 50% than at 75 and 95%. At 35 degrees C, L3 survived up to 6 weeks at 30%, 5 weeks at 50% and 1 week at 75 and 95% RH. At 16 and 25 degrees C and RH of 30 and 50%, over 70% of L3 were still alive at the end of experiments at the seventh week. At 25 degrees C, 40% of L3 survived for up to 5 weeks at 75% RH and 5% survived for 6 weeks at 95% RH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249247 TI - Comparison of three enzyme immunoassays for diagnosis of Dictyocaulus viviparus infection. AB - Three enzyme-linked immunosorbent assays (ELISA) that detect antibodies against Dictyocaulus viviparus in cattle were compared for sensitivity, specificity and seroconversion after primary infection. These assays were (i) an indirect ELISA with crude somatic antigens from adult D. viviparus (ca-ELISA), (ii) an indirect ELISA with purified antigens (sa-ELISA) isolated from somatic antigens of adult D. viviparus and (iii) a competition ELISA with antigen purified with anion chromatography in combination with monoclonal antibodies against D. viviparus. Sera from helminth-naive calves and sera from calves monospecifically infected with Ostertagia ostertagi, Cooperia oncophora, Nematodirus helvetianus, Ascaris suum or Fasciola hepatica were used to determine the specificity of the assays. Sera from calves and milk cows experimentally or naturally infected with D. viviparus, and from vaccinated calves, were used to test the sensitivity of the assays and to determine when the ELISAs detected seroconversion. The specificity of the competition and the sa-ELISA was 97%, whereas the specificity of the ca ELISA was 67%. The sensitivities of the sa-ELISA, the competition ELISA and the ca-ELISA were 97, 73 and 99%, respectively. All three assays detected seroconversion between 4 and 6 weeks after primary infection. None of the assays detected seroconversion in calves receiving lungworm vaccination. We conclude that of these three tests, the sa-ELISA can be used most beneficially to diagnose lungworm disease. PMID- 8249248 TI - Infection build-up and development of immunity in calves following primary Dictyocaulus viviparus infections of different levels at the beginning or in the middle of the grazing season. AB - An experiment was carried out to study the build-up of infection and development of immunity against Dictyocaulus viviparus infections in calves. Six groups of four calves were used. Groups 1-5 grazed on separate pastures and Group 6 served as a permanently housed control group. To simulate 'low' and 'very low' overwintered pasture infectivity, Groups 1 and 2 were infected experimentally six times with ten and two larvae, respectively, during the first 3 weeks on pasture in May. From the middle of July, Groups 2-4 were experimentally infected six times in 3 weeks with 10, 10 and 100 larvae, respectively, to simulate 'low' and 'moderate' pasture contamination by carriers. Group 5 served as a non-infected pasture control group. After housing of Groups 1-5 in October, all calves were treated with oxfendazole and challenged with 5000 larvae of D. viviparus to evaluate the development of immunity. The results showed a similar population build-up of lungworm infections in Groups 1-4. After the beginning of patency, 24 35 days after primary infection, an increase in faecal larval counts occurred 31 35 days later as a result of re-infection. For Group 2, this was only true when the infections with a total of 12 larvae in May were ignored, as these did not result in patent infections. Faecal larval counts were highest in Group 4, which was the only group with mild signs of lungworm disease occurring in the re infection period. The worm counts at necropsy showed the development of immunity in Groups 1-4 but not in Group 5. This is consistent with the fact that Group 5 did not develop patent lungworm infections throughout the grazing season. PMID- 8249249 TI - Lamb serum recognition of infective larvae and adult Haemonchus contortus antigens. AB - Sixteen- to eighteen-week-old lambs were infected with 2500 3rd stage larvae (L 3) of Haemonchus contortus or kept as uninfected controls. Two months later all animals were challenged with 5000 L-3 of this parasite. Soluble antigens of H. contortus L-3 and adult worms were analyzed by SDS-PAGE and Western blotting during experimental infection and challenge. Antigens from both sources, particularly of low molecular weight under reducing conditions, were recognised by the pooled sera of infected lambs. No single L-3 antigen was recognised by all infected lambs, whereas two peptides having around 25 and 26 kDa from adults were recognised by infected animals during the patency and could be of potential use in the diagnosis of lamb haemonchosis. PMID- 8249250 TI - Laboratory selection for ivermectin resistance in Haemonchus contortus. AB - A strain of Haemonchus contortus, susceptible to ivermectin (IVM), was submitted to a selection procedure using IVM at 0.02 mg kg-1 during nine successive passages in lambs. Treatment reduced worm egg counts (epg) by more than 99% up to the seventh passage, but thereafter there was no significant reduction in epg. The efficacy of ivermectin against the ninth selected generation compared with the original susceptible strain was assessed. The results showed that animals infected with the selected strain and treated with IVM did not show any significant reduction (P > 0.05) in epg compared with a 99.9% reduction in the susceptible group. At necropsy, post-treatment worm burdens confirmed that the selection pressure applied to the original strain had been successful in stimulating resistance to IVM. PMID- 8249251 TI - Parasite spectrum and seasonal epidemiology of gastrointestinal nematodes of small ruminants in The Gambia. AB - One hundred and four post mortem examinations of randomly selected sheep (52) and goats (52) were carried out weekly from March 1990 to February 1991. Gastrointestinal parasites were identified and counted. A large spectrum of 16 helminth species was found in 101 (97%) infected animals. The nematodes infecting small ruminants were, in order of predominance: Trichostrongylus colubriformis (96% of all animals), Oesophagostomum columbianum (82%), Haemonchus contortus (67%), Strongyloides papillosus (55%), Gaigeria pachyscelis (38%), Cooperia spp. (49%) and Trichuris ovis (12%). Forty-five (43%) necropsies revealed an infection with Moniezia benedeni, Avitellina centripunctata or Stilesia globulosa. Infections with Schistosoma bovis and Paramphistomum spp. were rarely seen. Goats carried significantly lower worm burdens than sheep and were less often infected by tapeworms. Nematode worm burdens followed a similar seasonal pattern in both host species. While Trichostrongylus colubriformis was also recovered in high numbers in the mid dry season, other species such as H. contortus, Strongyloides papillosus and Oesophagostomum columbianum reached distinct peaks in the rainy season (July-October). Different survival strategies for the prolonged dry season were observed. Haemonchus contortus outlived the unfavourable climatic conditions as inhibited larvae in the abomasal mucosa, while the other nematode species survived as adults with a reduced fecundity. Egg production per adult worm specimen was found to depend heavily on the season and egg counts in the dry season (November-June) did not correspond to the size of worm burdens. Unexpectedly high worm burdens were recovered from adult and old animals. Investigations of young animals indicated that no reinfection can occur during the dry season. The findings are discussed with regard to their relevance for strategic control of gastrointestinal nematodes in small ruminants. PMID- 8249252 TI - The potential of nematophagous fungi to control the free-living stages of nematode parasites of sheep: screening studies. AB - Ninety-four species of fungi with known nematophagous activity were tested for their ability to reduce the number of infective larvae of sheep nematodes in faecal cultures, and also for their ability to produce nematode-attractant and nematocidal substances against these free-living stages under in vitro conditions. Reductions of infective larval numbers exceeding 80% were consistently recorded when 100-250 conidia g-1 faeces of various species from the genera Arthrobotrys, Geniculifera and Monacrosporium were used. Even concentrations as low as 10 conidia g-1 faeces resulted in a significant reduction in infective larval numbers compared to control cultures. This study demonstrates that whilst many fungal species exhibit nematophagous activity against a variety of free-living nematodes, few show efficient activity against the free-living stages of parasitic nematodes in the sheep faecal environment. For the most active, there were six species of Arthrobotrys, two species of Geniculifera and two species of Monacrosporium which showed comparable activity to the extensively studied species A. oligospora. PMID- 8249253 TI - Impact of natural infestation of Amblyomma variegatum on the liveweight gain of male Gudali cattle in Adamawa (Cameroon). AB - The effect of natural tick infestation on the liveweight gain (LWG) of male Gudali zebu cattle was studied throughout a year by comparing the performances of two herds, one of which was submitted to weekly acaricidal treatment and the other was left untreated against ticks. Six species of ticks were identified on the untreated animals: Amblyomma variegatum, Boophilus decoloratus, Rhipicephalus lunulatus, Rhipicephalus turanicus, Hyalomma nitidum and Hyalomma marginatum rufipes. Most of the losses observed in the untreated herd during the rainy season were due to A. variegatum, and the loss in LWG was estimated to be 55-76 g per engorged female A. variegatum. The infestation also leads to wounds and to lesions of dermatophilosis. There was an interval between the peak infestation by A. variegatum and the appearance of weight loss owing to them. The control of ticks on the Gudali zebu in Adamawa, during the months of high infestation by A. variegatum adults, is economically profitable. On the other hand, the performances of the two herds during the dry season were similar, showing that infestation by larvae and nymphs of A. variegatum has no impact on the zebu LWG, and that tick control during that period is not profitable. PMID- 8249254 TI - The prevalence of concurrent trypanosome and gastrointestinal nematode infections in west African dwarf sheep and goats in Nsukka area of eastern Nigeria. AB - The prevalence of concurrent nematode-trypanosome infections in traditionally reared West African Dwarf sheep and goats in eastern Nigeria was monitored over a 12-month period during 1987-1988. The most prevalent nematodes were Haemonchus contortus and Trichostrongylus colubriformis, which usually occurred together in all nematode infected animals. Their combined prevalence rates ranged from 90 to 100% throughout the year and they accounted for 66 to 98% of the total monthly worm burdens. Of the 107 animals examined 13.6% were infected with trypanosome species comprising Trypanosoma brucei (50%), Trypanosoma congolense (43%) and Trypanosoma vivax (36%). No clear seasonal pattern was observed in the prevalence of concurrent nematode-trypanosome infection but owing to the widespread prevalence of gastrointestinal nematode infections, all trypanosome infected animals were invariably infected with H. contortus and Trichostrongylus colubriformis. PMID- 8249255 TI - Distribution of the number of Eurytrema sp. eggs per gram of feces in naturally infected cattle. AB - Eurytrema sp. egg counts (epg) in the feces of naturally infected cattle were performed and the technique employed showed 94.2% probability of detecting positive cases of the infection with a single examination independently of the host parasite burden. It was also demonstrated that the epg of Eurytrema sp. follows a negative binomial distribution model and is characterized by its small magnitude. PMID- 8249256 TI - Stomach parasites of donkeys in Morocco: habitat and interspecific interactions. AB - One hundred and four donkeys originating from several regions of Morocco were necropsied during 1 year. The parasites in their stomachs were counted and locations determined in the cardiac, fundic and pyloric portions. Synergistic or antagonistic interactions between parasites were assessed by a principal component analysis. Gasterophilus intestinalis was only found in the cardiac portion and G. nasalis was mostly found in the pyloric portion. Habronema were principally encountered in the pyloric portion, whereas Trichostrongylus axei was found in the fundic portion. The parasites were synergistically associated in their habitat interaction, except for T. axei on the fundic portion which was antagonistic to other parasites. PMID- 8249257 TI - Relationship between alpha-aminolevulinic acid dehydratase activity, free erythrocyte protoporphyrin concentration and blood lead in calves from lead contaminated farm. AB - In order to investigate useful indicators of lead contamination, 108 Japanese Black calves from a lead-contaminated farm were used. The lead concentrations in blood (Ph-B), delta-aminolevulinic acid dehydratase (ALA-D) activities, and free erythrocyte protoporphyrin (FEP) concentrations were examined. A significant negative correlation was obtained between Pb-B concentrations and ALA-D activities (r = -0.621, p < 0.01). A significant positive correlation was obtained between Ph-B concentrations and FEP concentrations (r = 0.850, p < 0.01). The calves were divided by Pb-B concentrations (micrograms/100 ml) into 5 groups (A = < or = 30, B = 31 approximately 90, C = 91 approximately 150, D = 151 approximately 210, E = > or = 211) to observe the relation of ALA-D activities, FEP concentrations and ALA-D activities in group B (r = -0.706, p < 0.05). A significant positive correlation was obtained between Pb-B concentrations and FEP concentrations in groups A, B and D (A = r = 0.496, p < 0.01; B = r = 0.686, p < 0.01; D = r = 0.529, p < 0.05). These results indicate that FEP concentrations were good indicators of lead contamination. PMID- 8249258 TI - Effect of aflatoxin-contaminated feed on morbidity and residues in walleye fish. AB - The effect of aflatoxin in the feed on the morbidity and toxin residue in the musculature of walleye fish was studied. The fish were divided into 3 groups of 12 each; group 1 served as controls, groups 2 and 3 received feed treated with 50 or 100 ppb toxin for 30 d respectively. Fish were kept in glass aquaria, 4 fish/aquarium, with aerator and solids-waste filter system. Fish were observed for morbidity and mortality. Six fish from each group were sacrificed after 30 d and examined for gross pathological lesions and histopathological changes. In addition, fish musculature were analyzed for aflatoxin. One out of 16 (8%) of the fish fed 100 ppb aflatoxin in 48 h. Pale livers and significant degenerative changes were observed in both toxin-treated groups. Aflatoxin B1, G1 and G2 were detected in the musculature of both groups at a concentration of 5, 10, 15, 20 ppb respectively. After 2 w of aflatoxin withdrawal, no aflatoxin was detectable in the fish muscle. However, marked histopathological lesions were still observed. PMID- 8249259 TI - Screening of "Kargasok tea". IV: Studies of pathological effects in BALB/C mice and Wistar rats. AB - Forty-eight BALB/C mice (30 males, 18 females) and 18 male Wistar rats were given Kargasok tea in their drinking water. Zero, 15, 25, 50, 75 or 100% concentrations were administered to mice for 2 or 8 w while the rats were given 0, 15 or 50% concentrations of the beverage for 12 w. Neither the mice nor rat's organ/body weight ratios were significantly affected by treatment. Rats receiving 15 or 50% concentrations of the tea had nephropathy and non-suppurative necrosis of the duodenum, pancreas and intestine. No significant microscopic lesions were found in any treated mice even when given 100% concentration of the tea. These findings suggest species variation in susceptibility to toxicity from the tea. The tea may be a cause of renal failure, diabetes mellitus and malabsorption syndrome and may not be safe for human or animal consumption. PMID- 8249260 TI - The efficacy of different decontaminants in rats and pigs percutaneously poisoned with organophosphates. AB - The efficacies of clay or alcoholate as decontaminants and a polyethylene glycol + oxime N-octylpyridium-4-aldoxime bromide (OPAB) protective ointment were evaluated in rats and pigs percutaneously poisoned with O-ethyl S-2 diisopropylaminoethyl methylphosphonothioate (VX) and soman (GD) nerve gases. The use of protective ointment per se or combined with the decontaminants protected all rats poisoned with GD, regardless of the experimental procedure. No poisoning or fatalities were observed in pigs decontaminated 2 min later with clay or alcoholate, while the protective ointment delayed the onset of poisoning and even prevented death. The application of protective ointment, with or without the use of decontaminants, significantly postponed the onset of poisoning in animals contaminated with VX. There was no significant difference between procedures. Decontamination was very satisfactory and dependent on the duration of exposure, being somewhat more efficient if performed by 30 min after exposure. PMID- 8249261 TI - Determination of chloramphenicol residues in eggs by high performance liquid chromatography (HPLC). AB - A rapid and sensitive HPLC method for the determination of chloramphenicol (CAP) residues in albumin and yolk of eggs is described. Conditions of isolation and identification are presented in detail. The detection limit is 10 ppb and the linearity is very good up to 2500 ppb CAP. The recovery of the method is 82 and 77%, and the precision is 6.6 and 11.1% for the albumin and yolk determinations respectively. The method was tested to quantitate CAP residues in eggs of 5 laying hens to which CAP had been administered in feed at a concentration of 800 mg/kg. CAP was detected in albumin and yolk of eggs laid 3 and 8 days after the CAP administration, respectively. PMID- 8249262 TI - The cupruretic effect of two chelators following copper loading in sheep. AB - The cupruretic effect of 2 copper chelators, d-penicillamine and trientine, was assessed in 12 fistulated South African Mutton Merino rams of approximately 6 mo age following copper loading. Each animal received 20 mg CuSO4.5H(2)0/kg body mass as an 0.5% m/v aqueous solution, intraruminally, daily for 35 d. The animals were randomly assigned to either a d-penicillamine (n = 4) or trientine treatment group (n = 4) or an unmedicated control group (n = 4). A separate group of 3 rams were kept as non-copper-loaded controls. Urinary copper excretion was measured before and during treatment. All the sheep were housed individually or placed intermittently on steel metabolic crates to facilitate urine collection. At the end of the trial the animals were euthanatized and specimens of organs collected for determination of copper concentrations. d-Penicillamine significantly (p < 0.05) increased urinary copper excretion. Trientine failed to increase copper excretion in the urine when compared to the unmedicated and non-copper-loaded control groups. PMID- 8249263 TI - Synergistic effects of monensin in combination with permethrin or neomycin on neuronal activity. AB - Drug combinations have the potential to produce novel and unpredictable responses on nervous tissue. This study was designed to test the hypothesis that the effects of combinations of monensin (an ionophore antibiotic) and either neomycin (an aminoglycoside antibiotic) or permethrin (synthetic pyrethroid) are synergistic. Effects of the drug combinations upon the electrical properties and membrane activities of an in vitro sensory neuron preparation were found to be greater than expected from addition of the effects of the same drugs acting individually, indicating synergism and thus supporting the hypothesis. It was concluded that drugs acting at different neuronal membrane sites and applied in combination produce unpredictable responses. Such drug combinations behave as if they were novel drugs. PMID- 8249264 TI - The efficacy of different formulations of protective-decontaminating ointments based on polyethylene glycol in rats percutaneously poisoned with highly toxic organophosphates. AB - Protective-decontaminating ointments based on polyethylene glycols (PEG) of different molecular weights were tested alone or with oxime N-octylpyridinium-4 aldoxime bromide, against percutaneous poisoning by the nerve gases 0-ethyl S-2 diisopropyl methylphosphorothiolate and soman in rats after 60 min exposure. PEG based ointments consisting of higher PEG combined with the oxime proved the most efficient for both poisons. The combination of lower and medium PEG was somewhat less efficient, while the formulation consisting of mostly lower PEG exerted protective effects only in soman poisoning. PMID- 8249265 TI - Characterization of particles, ammonia and endotoxin in swine confinement operations. AB - To define the atmospheres swine confinement operations, we measured concentrations of total and respirable dust particles, ammonia and endotoxin in the nursery and grower areas of 4 swine confinement houses. Increased ventilation in spring-summer relative to that in winter reduced concentrations of large dust particles more rapidly than it did smaller particles or ammonia. The greater decrease in large particles correlating to increased room air velocity may reflect larger particles' momentum causing impaction on surfaces. There was significant spatial variation in the concentration of airborne endotoxin within individual swine rooms and pens reflecting different mixing of large feed and smaller manure particles. Smaller particles had 4-fold higher concentrations of endotoxin than did larger particles, suggesting they had higher fecal material concentrations. Total airborne endotoxin and total suspended particulates correlated to the fraction of functional endotoxin contained in large particles, suggesting that small particles (0.5-2.0 micrometers) collide with large particles (50 micrometers). These data suggest that large non-respirable particles remove smaller respirable particles from indoor atmospheres due to kinematic coagulation. PMID- 8249266 TI - Teratogenic effects of a mixture of scopolamine and hyoscyamine in chick embryos. AB - The effect of a mixture of scopolamine and hyoscyamine (98:2) on developing chick embryos was investigated. The embryos were exposed at 2 stages of development to various doses of this mixture via yolk sac injection. This resulted in deaths at different stages of prenatal development, as well as various malformations and reduction deformities. These findings suggest that the studied mixture has a teratogenic effect when injected into the yolk sac of developing chick embryos. PMID- 8249267 TI - Vicia tetrasperma (four-seeded vetch) ingestion by a 3-year-old child. AB - We report an ingestion of Vicia tetrasperma (four-seeded vetch), initially misidentified as crown vetch (Coronilla varia). Vicia tetrasperma is not listed in POISINDEX; little is known of its toxic effects and we found no published human case reports. The child suffered only minor gastrointestinal effects which lasted a few hours and had no residual effects upon 24- and 96-hour followups. PMID- 8249268 TI - Evolution of fluoride concentrations in cattle and grass following a volcanic eruption. AB - This study determined fluoride concentrations of forage, water and bovine serum 2 y after the cessation of a volcanic eruption. Three farms within the polluted area were selected, and water and green grass were collected from April 1990 to February 1992. Samples of hay were collected twice from each farm. Five cows exposed during the 13-mo eruption and 6 young bulls not exposed during the volcanic eruption had blood collected monthly or bimonthly. Serum fluoride concentrations and alkaline phosphatase activities were determined and dental conditions observed in all animals. Grass fluoride remained low except during spring of the second year; high fluoride was in the hay. Serum fluoride of the cows remained near acceptable reference values, although some cows had higher values; serum fluoride of the bulls markedly increased during the spring of the second year. Severe incisor attrition occurred in the cows, while the young bulls had less severely eroded teeth. PMID- 8249269 TI - Successful therapy of iron intoxication in pregnancy with intravenous deferoxamine and whole bowel irrigation. AB - We describe the successful treatment of a severely iron-poisoned adult patient in week 26 of gestation with 10.2 g deferoxamine administered iv over 14 h and whole bowel irrigation (2 L/h of polyethylene glycol-electrolyte solution/nasogastric tube for 12 h) with a good maternal outcome and no adverse effects on the fetus. PMID- 8249270 TI - Grand mal seizure and clobutinol overdose. AB - We report the case of a previously healthy girl who developed a grand mal seizure after an inappropriate dose of clobutinol and had a good response to iv diazepam. PMID- 8249271 TI - A review of thallium toxicity. AB - Thallium (Tl) is one of the most toxic of the heavy metals. Its continued use as a rodenticide in many developing countries and its increasing use in an expanding number of new technologies raise concerns about exposure risk to animals and humans. Because Tl and potassium (K) have the same charge and similar ionic radii, Tl follows K distribution pathways and alters and number of K-dependent processes. Possible toxic mechanisms of Tl include ligand formation with protein sulfhydryl groups, inhibition of cellular respiration, interaction with riboflavin and riboflavin-based cofactors, and disruption of calcium homeostasis. The principal clinical features of thallotoxicosis are gastroenteritis, peripheral neuropathy of unknown etiology, and alopecia. The presence of elevated Tl levels in the urine or other biologic materials confirms the diagnosis of Tl poisoning. Treatment with prussian blue (or activated charcoal) will interrupt the enterohepatic cycling of Tl, thus enhancing fecal elimination of the metal. Forced diuresis with potassium loading will increase the renal clearance of Tl, but should be used cautiously because neurologic and cardiovascular symptom may be exacerbated. If recognized and treated early, Tl poisoning carries a favorable prognosis for full recovery. PMID- 8249272 TI - Fescue toxicosis and its impact on animal agriculture. AB - Studies of fescue toxicosis have identified several classes of plant/fungal alkaloids that may be responsible for the toxicosis problem of endophyte (Acremonium coenophialum) infected tall fescue (Festuca arundinacea). This review explores the interaction of these alkaloids with the animal's system and attempts to explain the various anomalies of fescue toxicosis through discussion of both in vitro and in vivo experiments. PMID- 8249273 TI - Problems associated with the use of common names in the identification of poisonous plants. AB - Barriers in communication among professional botanists, physicians and the general populous developed from a disparity in the use of botanical nomenclature. Universally accepted scientific names from plants affords a common ground of understanding among botanists throughout the world. A general phobia with unfamiliar latinized scientific nomenclature has proliferated the use of common or trivial names. These names, however, can vary with geographic region, language, or individual preference often and thus, can lead to misidentification or a delay in the proper identification of a toxic plant. PMID- 8249274 TI - Illiteracy: a contributing factor to poisoning. AB - It is estimated that over 60 million Americans (1/3 of the adult population) are functionally or marginally illiterate. To recognize and gain an understanding of illiteracy and its impact on poisoning, we determined if the illiterate in our community could recognize potential poisons. A prospective study involving 29 male and 21 female adult illiterates was undertaken. A personal interview was conducted to determine their ability to purchase medication and household products, their understanding of the uses and associated dangers of medications, and their manner of storage of these products in their homes. Of the participants, 66% were at reading levels of 0-3rd grade and 34% were at 3rd through 6th grade reading level. Each participant was shown 3 separate products and asked to distinguish and interpret caution statements and directions. In the 0-3rd grade group, 30% were unable to identify any of the products and none could explain the cautions or directions. In the second group (3rd-6th grade), all were able to identify the products and 76% could explain the cautions; everyone in this group correctly read the directions. We concluded that a large percentage of the adult population are potential poisoning victims due to their inability to read and comprehend label instructions. Poison Centers should recognize illiteracy as a contributing factor in poisonings and consider education and prevention programs for this segment of our population. PMID- 8249275 TI - DNA sequence analysis of conserved and unique regions of swinepox virus: identification of genetic elements supporting phenotypic observations including a novel G protein-coupled receptor homologue. AB - Swinepox virus (SPV) contains a double-stranded cross-linked linear DNA genome of approximately 175 kilobase pairs with terminal inverted repetitions (TIRs) of 4.3 kb. The nucleotide sequence was determined for fragments from several regions of the genome including a 2.85-kb fragment from the central potentially conserved portion and two fragments within the presumed variable near-terminal regions which tend to be unique to a given poxvirus. The core sequence contains one partial and two complete open reading frames that are highly conserved and colinear with three contiguous ORFs within the HindIII D fragment of vaccinia virus (VV). The two near-terminal fragments, encompassing 14.2 and 3.6 kb, are respectively located 2.1 kb internal to the left and right cross-linked termini of the DNA and span the TIR junctions. The sequences encode 25 open reading frames including numerous proteins predicted to be membrane-bound or secreted in infected cells. Several ORFs unique to SPV were identified that may be involved in cell attachment, immune modulation, and pathogenesis including a novel poxvirus G protein-coupled receptor. In addition, several polypeptides encoded within the near-terminal regions of vaccinia virus DNA that function as host range or virulence factors are lacking within this region of swinepox virus including the VV growth factor, complement-binding protein, and ORFs C7L and K1L, associated with host range. The lack of these functional homologues could explain the characteristic attenuated phenotype and limited host range of SPV. PMID- 8249276 TI - Two-codon insertion mutations of the HBx define two separate regions necessary for its trans-activation function. AB - A panel of mutants of the hepatitis B virus X gene (HBx) was constructed by oligonucleotide-directed insertion of two codons (Arg Pro) at 10- or 20-amino acid intervals along the entire gene. These mutants were tested in transiently transfected HepG2 cells for their effects on HBx trans-activation of an AP1-CAT reporter plasmid. The effects of HBx mutations on the mRNA and protein stability were also determined. Our results reveal two separate internal domains of the HBx, one around amino acid residue 68 and the other between residues 110 and 139, that are necessary for its activity. Mutations in the amino terminus (to position 45), between the two necessary domains and in the extreme carboxyl terminal portion of HBx, have little or no effect on its activity. PMID- 8249277 TI - Large T-antigen and sequences within the regulatory region of JC virus both contribute to the features of JC virus DNA replication. AB - The requirements for the DNA replication of the human papovavirus JC were analyzed using JC T-antigen as well as the T-antigens of the related viruses SV40 and BK. With all three T-antigens, the boundary of the core origin mapped on the early side to position 5093 of the viral genome. In conjunction with earlier studies, the core origin of DNA replication was therefore defined as a 68-bp region which, similar to the SV40 core origin, contains three major structural elements, early palindrome, T-antigen binding site II, and A/T-rich tract. Replication was stimulated by sequences flanking the core origin on the early side. Specifically, the stimulating sequences on the early side were identified as T-antigen binding site I. The degree to which flanking sequences were able to stimulate viral DNA replication was dependent on the T-antigen used in the experiment, with JC T-antigen relying most and BK T-antigen relying least on the flanking sequences. SV40 T-antigen showed an intermediate dependence. The same hierarchy was observed when replication activities were compared. BK T-antigen was more active in replicating DNA than SV40 T-antigen, which in turn was more effective than JC T-antigen. Dependence on flanking sequences is, thus, inversely correlated to the replicating activity of the respective T-antigen, showing that, in addition to the origin, the T-antigen contributes to the characteristics of JC virus DNA replication. PMID- 8249278 TI - Endonexin II, present on human liver plasma membranes, is a specific binding protein of small hepatitis B virus (HBV) envelope protein. AB - Binding of viral envelope proteins to specific receptors on human hepatocytes is considered to be an important step in HBV infection. In this study, we demonstrate that a 34-kDa human liver plasma membrane protein specifically binds to small HBsAg in a Ca(2+)-dependent manner. By partial amino acid sequence analysis of preparatively isolated 34-kDa protein comigrating with HBsAg-binding protein obtained from binding assay on IEF/SDS-PAGE, we have identified this HBsAg-binding protein as Endonexin II (E-II). Native human liver E-II inhibits binding of HBsAg to intact human hepatocytes and shows specific binding to small HBsAg. This binding can be inhibited by human liver plasma membrane proteins, recombinant E-II, or anti-E-II antibodies. Despite 90% sequence homology, rat liver E-II does not bind to small HBsAg and does not inhibit significantly (less than 20%) binding of HBsAg to intact hepatocytes. Cross-linking of small HBsAg and radiolabeled human liver E-II resulted in a specific additional protein complex on PAGE with an apparent molecular weight of 90 kDa, corresponding to a complex of E-II and small HBsAg with a ratio of 2 to 1 or 1 to 2. These findings indicate that E-II, found in human liver, is a specific HBsAg-binding protein and might play an important role in the initiation of HBV infection. PMID- 8249279 TI - A reassortant H1N1 influenza A virus caused fatal epizootics among camels in Mongolia. AB - In the autumn of 1979 a severe influenza epizootic started among camels in Mongolia (Lvov et al., 1982; Viprosi Virusol. 27, 401-405.) Between 1980 and 1983 13 independent isolates of H1N1 viruses were obtained from diseased camels, which were virtually indistinguishable from the human A/USSR/90/77 strain by serological means. Two hundred and seventy-one samples of camel sera collected between 1978 and 1983 contained antibodies against the human A/USSR/90/77 isolate. After experimental infection of camels with some of these isolates, the animals developed similar symptoms as those found during natural infection: coughing, bronchitis, fever, discharge from nose and eyes. A genetic sequence analysis revealed that among the eight segments (genes) the PB1, HA, and NA genes were almost identical with allelic genes of the USSR/77 strain, and the PB2, PA, NP, M, and NS genes were almost identical with those of the A/PR/8/34 strain. PMID- 8249280 TI - High rate of infection with the human T-cell leukemia retrovirus type II in four Indian populations of Argentina. AB - Sera from 215 non-drug-injecting Toba and Mataco-Mataguayo pure Indians belonging to four communities in northern Argentina were examined using assays that allow differentiation between reactivities due to type-specific antigens of the human T cell leukemia/lymphoma virus (HTLV). Three of these populations have very little contact with non-Indian groups and reside in remote, isolated areas. HTLV-II type specific seroreactivity was present in 24 (13.7%) of the 175 Indians older than 13 years of age and in none of the 40 who were of younger ages. None of the Indians had antibodies reacting with HTLV-I type-specific antigen. Seroreactivity was more prevalent and appeared at younger ages in females than in males. The majority of the HTLV-II-seropositive Indians belonged to the more isolated communities. The seroprevalences among the Tobas and Mataco-Mataguayo Indians were comparable. With the exception of a Toba who was positive in a test for Treponema pallidum, no serological evidence of sexually transmitted infections with this spirochete, hepatitis B virus, hepatitis C virus, and human immunodeficiency virus was found among the Indians tested. None of the 55 non Indian people tested in the region showed HTLV-II type-specific seroreactivity. PCR analysis of DNA isolated from peripheral blood lymphocytes of seropositive Indians confirmed that the virus present in these populations is HTLV-II. Sequence analysis of PCR-amplified genomic segments showed that the virus belongs to the HTLV-II subtype which has been found to be endemic in other Paleo-American Indians. PMID- 8249281 TI - Decreased reporter gene expression during latent infection with HSV LAT promoter constructs. AB - The latency-associated transcripts (LAT), which code from an 8.5 kb segment of the internal repeat region of the HSV genome, are the only viral transcripts that are present during HSV latent infection. However, little is known about the relative contribution of promoter activity, degradative processes, and elements or regions affecting long term expression of these transcripts in latently infected neurons. To begin to address this question we investigated LAT promoter activity during acute and latent infection. Mouse footpads were infected with KOS/62-3, an engineered herpes simplex virus in which both copies of the LAT promoter are used to drive expression of the Escherichia coli lac Z gene. Four days post-inoculation (p.i.) abundant beta-galactosidase (beta-gal) protein and transcripts were present within ganglionic neurons as assayed by enzyme histochemistry and in situ hybridization. In contrast, by Day 21 (at which time a latent infection had been established) no beta-gal transcripts were present in infected ganglia, even when assayed by the polymerase chain reaction (PCR). These findings indicate a significant drop in LAT promoter activity between Day 4 and Day 21 p.i. To provide confirmatory evidence for this conclusion we infected mice with a second viral construct, KOS/67-7, in which the LAT promoter was used to drive expression of the nerve growth factor (NGF) gene. Four days p.i., abundant NGF antigen and transcripts were present in infected ganglionic neurons, but no evidence of transcription of the cloned NGF gene could be found in latently infected ganglia. Our findings suggest that LAT promoter activity is severely restricted during the latent phase of ganglionic infection. PMID- 8249282 TI - Sequence and functional analysis of the human adenovirus type 7 E3-gp19K protein from 17 clinical isolates. AB - The adenovirus (Ad) early region 3 (E3) glycoprotein of 19K (gp19K) binds major histocompatibility (MHC) class I antigens in the endoplasmic reticulum (ER), and the gp19K-class I complex is retained in the ER through an ER retention signal at the C-terminus of gp19K. This retention of class I antigens blocks cytolysis of gp19K-expressing cells by cytotoxic T lymphocytes (CTL). Animal models infected with Ad mutants lacking gp19K support a role for gp19K in counteracting a CTL response. Gp19K binds with different avidities to different class I antigens, and portions of the gp19K sequence are highly variable among Ad serotypes in different subgroups (Ad3, 11, and Ad35 in subgroup B; Ad2 and Ad5 in subgroup C); this raises the possibility that certain human individuals may be more susceptible to productive or persistent infection by particular serotypes of Ad, depending on the haplotype of the individual and the type of Ad. To begin to address this possibility, the gp19K gene from 17 very diverse Ad7 (subgroup B) clinical isolates was amplified by the polymerase chain reaction, and the DNA sequences were determined. The Ad7 gp19K sequence was 98% identical to that of Ad3. Surprisingly, we found complete conservation of the amino acid sequence of gp19K from all but one of the clinical isolates; one isolate had a conservative Ala to Val substitution. Gp19K from Ad7 clinical isolates representing distinct Ad7 genotypes co-immunoprecipitated with class I antigens. Our data indicate that there is very strong evolutionary pressure to maintain the sequence of gp19K in Ad7. The only known function for gp19K from different Ad serotypes is binding to class I antigens. It is interesting to consider, therefore, what selective pressure operates to maintain the sequence of gp19K among serotypes within a subgroup, and yet allows for very significant divergence in the sequence of gp19K among serotypes in different subgroups. The possible role of MHC class I antigens in this selection process is discussed. PMID- 8249283 TI - Specific interaction of the human immunodeficiency virus Tat proteins with a cellular protein kinase. AB - The human immunodeficiency virus types 1 and 2 (HIV-1 and HIV-2) Tat proteins are related transcriptional activators whose effects are likely to be mediated by a cellular factor. Using an in vitro kinase assay, we have shown that the Tat protein of HIV-2 and the activation domain of the Tat protein of HIV-1 specifically bind to a cellular protein kinase. Mutations in Tat that abolish transactivation activity in vivo abrogate the ability of the mutants to bind to the kinase in vitro. This is the first demonstration of a cellular factor that binds to Tat that is specific for a functional activation domain of Tat and that displays a biochemical activity. Additionally, we show that the Tat protein of HIV-2 serves as a substrate of the kinase in vitro. Consistent with the in vitro results, the Tat protein of HIV-2 interacts with a cellular kinase in HIV-2 Tat transfected cells and is phosphorylated in vivo. These results suggest that a cellular serine/threonine kinase may act as a mediator of Tat function. PMID- 8249284 TI - RNA-dependent cleavage of VP0 capsid protein in provirions of hepatitis A virus. AB - Stable provirions of hepatitis A virus containing up to 62% VP0 were purified from infected BS-C-1 cells by sucrose density gradient ultracentrifugation, and conversion of these provirions to virions through maturation cleavage of VP0 capsid protein was demonstrated. VP0 cleavage was slow but linear over 7 days at 37 degrees, with mature virions containing between 3 and 7 copies of VP0 in separate experiments. Cleavage of approximately 25% of VP0 molecules (15 copies) was accompanied by a twofold increase in specific infectivity. Particles with reduced levels of VP0 were observed to sediment more rapidly in sucrose than VP0 rich provirions, reflecting conformational changes in the particles. The kinetics and temperature-dependence of VP0 cleavage further suggest that such conformational changes accompanying VP0 cleavage are necessary for the formation of subsequent catalytic sites. PMID- 8249285 TI - Transmission of human immunodeficiency virus type 1 (HIV-1) from mother to child correlates with viral phenotype. AB - The aim of this study was to investigate if the risk of mother-to-child transmission of human immunodeficiency virus type 1 (HIV-1) is influenced by the biological phenotype of the mother's virus. Virus isolates from 30 HIV-1 infected mothers and 12 infected children born to these mothers were analyzed for replication on several cell lines (Jurkat-tat, Jurkat, CEM, U937 clone 2, and MT 2). We show that mothers who harbor virus able to replicate in cell lines (rapid/high virus) have a significantly higher risk to infect their children than mothers with slow/low virus (P = 0.017). Children born to mothers with rapid/high viruses can be infected by slow/low as well as rapid/high viruses, while mothers with slow/low virus appear to transmit slow/low virus in every case. Our study shows that the biological phenotype of the mother's virus may serve as a complementary marker to CD4+ lymphocyte counts and p24 antigenemia in predicting the risk of transmission of HIV-1 to the child. PMID- 8249286 TI - Inhibition of heterologous DNA replication by the MVMp nonstructural NS-1 protein: identification of a target sequence. AB - The nonstructural protein NS-1 of minute virus of mice (MVMp), an autonomous parvovirus, trans-inhibits the replication of a chimeric plasmid containing the SV40 origin of replication (ori) embedded in the MVMp genome. It appears that a 157-bp 5' proximal sequence of MVMp DNA is sufficient, in the presence of NS-1, to cause the inhibition of DNA replication driven by the SV40 ori placed on the same molecule. This effect is not dependent on the orientation of the MVMp target sequence and results from both a reduced level of utilization of SV40 ori and the blockage of progressing replication forks at the level of the target. Furthermore, replication driven by Epstein-Barr virus origin (oriP) is trans inhibited by MVMp but this inhibition does not require the presence of parvoviral sequences in cis. On the basis of sequence homologies between EBV oriP and MVMp 5' terminal sequence, it is proposed that the direct or indirect interaction of NS-1 with parvovirus-like sequences present in heterologous viral and possibly also cellular genomes may result in an inhibition of DNA replication. PMID- 8249287 TI - Function of the mouse Mx1 protein is inhibited by overexpression of the PB2 protein of influenza virus. AB - The interferon-induced murine Mx1 protein possesses an intrinsic antiviral activity with selectivity for influenza viruses. Mx1 accumulates in the nucleus and inhibits the replication of influenza virus at the level of primary transcription. Simultaneous overexpression of the three influenza virus polymerase subunits via recombinant vaccinia virus vectors can titrate out the inhibitory action of Mx1 as determined by the amplification of a transfected recombinant viral reporter gene. A low degree of neutralization was also observed, when PB2 was overexpressed alone [Huang, T., Pavlovic, J., Staeheli, P., and Krystal, M. (1992) J. Virol. 66, 4154-4160]. We now employed a much simpler experimental setting which allowed us to directly measure the effect of PB2 on the antiviral activity of Mx1. We stably transfected a cell line derived from an A2G mouse (homozygous for a functional Mx1 gene) with expression vectors coding for cDNAs of the influenza virus polymerase subunits PB1 and PB2 or of the nucleoprotein (NP). Cells coexpressing Mx1 and PB1 or NP remained resistant to influenza virus infection whereas cells coexpressing Mx1 and PB2 became sensitive to influenza virus infection. The degree of neutralization of Mx1 activity by PB2 was dependent on the Mx1 concentration in the cell. Immunofluorescence analysis revealed that the nuclear localization of Mx1 and PB2 overlapped to a great extent. These findings support the view that Mx1 exerts its antiviral activity by interfering with the function of the influenza virus polymerase subunit PB2. PMID- 8249288 TI - Dynamics of genome change in the E2/NS1 region of hepatitis C virus in vivo. AB - To substantiate and extend the quasispecies model of hepatitis C virus (HCV), we made a pairwise comparison in the nucleotide and deduced amino acid sequences for multiple recombinant clones of the E2/NS1 region, which derived from each blood sample taken from five patients of subtype II or subtype III infection at different stages. Sequence heterogeneity among the clones was generally high. The heterogeneity, however, changed temporally and appeared to be significantly lowered after interferon therapy. The temporal fluctuation involved selection of particular amino acids at particular positions, which had represented only a minor fraction or had been absent in the previous clones. Evolution of defective viruses was featured by most of the cases, the proportion of which also fluctuated temporally and was extremely high at a certain stage in one of the patients. A comparison was also made among the clones from different stages of each patient and revealed highly divergent clones. An extreme case of subtype III infection showed sequence differences well beyond those among chronologically and geographically different isolates of subtype I or subtype II, which were defined by the same calculation of available sequence data. These features of HCV genome suggest that the virus could circulate as an extremely heterogeneous population including defective viruses and that this heterogeneity lends itself to selection pressures including interferon therapy and host immune response. Our results also indicate an absolute need of population based approach in HCV genetics. PMID- 8249289 TI - Teratogenic effects of neonatal arenavirus infection on the developing rat cerebellum are abrogated by passive immunotherapy. AB - The effects of viral infection on the developing nervous system and the potential of passive immunotherapy to protect against infection were examined. When 4-day old Lewis rats were injected intracerebrally with lymphocytic choriomeningitis virus (LCMV) the majority of stem cells within the external granular layer of the developing cerebellum became infected. The infection progressed to the molecular layer, internal granular layer, and the Purkinje cells. By 15 days postinfection the molecular and internal granular layers of LCMV-infected cerebella were noticeably thinner than those in the controls and the individual folia were smaller. Neurons remained infected for up to 40 days as determined by immunohistochemistry. However, in rats treated with rat monoclonal anti-LCMV antibodies the staining was limited to the cells of ependyma and choroid plexus and was not detectable by 15 days postinfection. Macroscopically the infection resulted in pronounced hypoplasia, with the cerebella of 21-day-old LCMV-infected rats weighing 52 +/- 10 mg compared with 159 +/- 30 mg for control rats. Antibody treated rats exhibited normal cerebellar size and development. Neutralizing antibodies specific for the viral GP-1 glycoprotein were protective but nucleoprotein-specific antibodies were not. Furthermore, suckling rat pups born of and nursed by LCMV-immune mothers were spared from cerebellar disease following neonatal infection. These results suggest that passive immunotherapy of neonates can provide effective protection against teratogenic effects of neonatal viral infection on the developing CNS. PMID- 8249290 TI - HF1 and HF2: novel bacteriophages of halophilic archaea. AB - Two novel halophilic archaebacterial bacteriophages, HF1 and HF2, were isolated from an Australian solar saltern. They were morphologically identical with icosahedral-shaped heads (diameter 58 nm) and contractile tails (length 94 nm). Other similarities included sensitivity to reduced ionic conditions, similar protein profiles by SDS-PAGE, and dsDNA genomes of identical size (73.5 kbp) with analogous restriction patterns. DNA-DNA hybridization data showed the two phages to be closely related. HF1 has a broad host-range, infecting members of three halobacterial genera including Halobacterium salinarium and the genetically well characterized strain Haloferax volcanii WFD11. Mutants showing increased plating efficiency on alternative hosts were readily selectable. By contrast, HF2 showed a limited host range, confined to the closely related dam-methylated strains Ch2 and H. saccharovorum. PMID- 8249291 TI - Purification and biochemical characterization of the protein kinase encoded by the US3 gene of herpes simplex virus type 2. AB - Post-ribosomal cytoplasmic fractions from Vero cells mock-infected or infected with wild-type herpes simplex virus type 2 (HSV-2) or a US3 gene-disrupted mutant of HSV-2 were fractionated with DEAE-cellulose chromatography, and the peak fraction of the protein kinase which was detectable only in the extract of wild type virus-infected cells was subjected to successive chromatography. The enzyme was purified more than 1000-fold from the post-ribosomal supernatant, and the final preparation contained one major protein of apparent molecular weight 66 kilodalton (K), which was phosphorylated in the autophosphorylation reaction. Western blotting analysis showed that antibodies to an synthetic peptide corresponding to the 15 amino acids of the predicted HSV-2 US3 protein sequence strongly reacted with a 66 K protein in the enzyme fractions. On Superose 12 HR chromatography, the protein kinase activity was eluted as a single major peak at a position corresponding to an apparent molecular mass of approximately 60 K. These results suggest that the 66 K protein is the protein kinase encoded by the US3 gene of HSV-2 and that it acts as a monomer. The HSV-2 protein kinase was relatively resistant to high concentrations of salt, but KCl above 400 mM exerted a significant inhibitory effect. When the substrate specificity was investigated using synthetic oligopeptides, the peptides containing arginyl residues on the amino-terminal side of the target seryl residue were found to be the best substrates for the protein kinase. However, the replacement of the seryl residue to threonine markedly reduced the rate of phosphorylation by this enzyme, suggesting that threonine is a poor phosphate acceptor of the protein kinase. The enzyme was resistant to heparin, a potent inhibitor of casein kinase II, but was moderately sensitive to H-9 (N-(2-aminoethyl)-5-isoquinolinesulfonamide dihydrochloride), a potent inhibitor of cyclic nucleotide-dependent protein kinases and protein kinase C. Quercetin, a bioflavonoid, also inhibited the protein kinase and the inhibitory effect was competitive towards ATP (Ki = 10 microM). The results indicate that the biochemical properties of the HSV-2 US3 protein kinase are very similar to those of the HSV-1 counterpart and pseudorabies virus-encoded 38-kDa protein kinase, but are different from those in several respects. PMID- 8249292 TI - Partial purification and characterization of foxtail mosaic potexvirus RNA dependent RNA polymerase. AB - The RNA-dependent RNA polymerase (RdRp) of foxtail mosaic virus (FMV) was partially purified from infected leaves of Chenopodium quinoa. The membrane fraction of crude plant extracts contained most of the FMV RdRp activity. Additional purification was obtained by solubilization of the RdRp using KCl and dodecyl-sucrose and by centrifugation through a glycerol gradient. The RNA template endogenous to RdRp preparations could be removed using micrococcal nuclease but the resulting fraction was unable to copy added template purified from FMV virions. However, supplementation of fractions containing RdRp activity with FMV RNA resulted in a significant decrease in the level of RNA synthesis. This effect was specific to potexviral RNAs since a similar interference was also observed with clover yellow mosaic virus RNA but not with brome mosaic virus RNA or yeast RNA. RNA transcripts corresponding to various regions of the FMV genome were tested for their ability to inhibit RNA synthesis on endogenous template. The simultaneous presence of both 5' and 3' terminal regions of the viral genome was necessary to interfere with RNA synthesis suggesting that this inhibition resulted from competition for the binding of component(s) of the RdRp complex. PMID- 8249293 TI - Identification of genes encoding late expression factors located between 56.0 and 65.4 map units of the Autographa californica nuclear polyhedrosis virus genome. AB - Using a previously developed method that allows the identification of Autographa californica nuclear polyhedrosis virus (AcMNPV) genes which stimulate transient expression from a late and a very late viral promoter [Passarelli and Miller, J. Virol., 67, 2149-2158 (1993)], we have identified three genes between 56.0 and 65.4 map units of the AcMNPV genome involved in expression from a late and a very late promoter but not from an early viral promoter. One gene, p143, was previously shown to be essential for viral DNA replication and shares sequence motifs with DNA helicases [Lu and Carstens, Virology, 181, 336-347 (1991)]. The second gene, previously sequenced and originally referred to as open reading frame 6 (herein renamed late expression factor-5 [lef-5]), was located just downstream of the 6.9 kilodalton core protein gene, p6.9. The third gene, late expression factor-4 (lef-4), was defined and sequenced. The lef-4 gene was located immediately upstream of, and in opposite orientation to, the major capsid protein gene, vp39. The position and direction of lef-4 appeared to be conserved in the Orgyia pseudotsugata and Lymantria dispar nuclear polyhedrosis viruses. The gene product of lef-4, LEF-4, is predicted to be an acidic polypeptide (pl 4.91) of 464 amino acids in length with a molecular mass of 53,913 daltons. PMID- 8249294 TI - Identification and characterization of a putative origin of DNA replication in the genome of a baculovirus pathogenic for Orgyia pseudotsugata. AB - A four-kilobase (kb) region (HindIII-N, map units 7.0-11.3) of the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (OpMNPV) genome was found to contain sequences that conferred upon plasmids the ability to undergo infection-dependent replication. The plasmid DNA appeared to be replicated to form high molecular weight multimers. Plasmids with deletions of up to 1.8 kb from either end of the HindIII-N region were replication competent. However, a discrete sequence, contained within the region bracketed by the deletions, capable of specifying replication was not identified. No evidence for sequence homology was found between the OpMNPV HindIII-N region and regions elsewhere in the OpMNPV genome or to putative Autographa californica MNPV (AcMNPV) replication origins. Origin-dependent plasmid replication was shown to require the presence of the OpMNPV DNA polymerase gene. The OpMNPV origin replicated poorly in AcMNPV infected Spodoptera frugiperda cells and, conversely, a putative AcMNPV origin (hr2) replicated at low levels in OpMNPV-infected Lymantria dispar cells. PMID- 8249295 TI - Generation of reovirus core-like particles in cells infected with hybrid vaccinia viruses that express genome segments L1, L2, L3, and S2. AB - When mouse L fibroblasts are infected with various combinations of recombinant vaccinia viruses possessing thymidine kinase (TK) genes with inserted reovirus genes that encode core components, particles are formed that closely resemble reovirus cores. In cells infected with vaccinia viruses that express reovirus proteins lambda 1 and sigma 2, particles are formed that are very similar to reovirus core shells; if, in addition, the cells are also infected with vaccinia virus that expresses protein lambda 2, particles are formed that also possess the characteristic icosahedrally located projections/spikes that are present on reovirus cores. If, in either case, the cells are also infected with vaccinia virus that expresses the reovirus RNA polymerase, protein lambda 3, the resultant particles are morphologically identical with those formed in its absence, but also contain protein lambda 3. PMID- 8249296 TI - Cell fusion studies identified multiple cellular factors involved in mouse hepatitis virus entry. AB - We have previously reported that certain murine cell lines are susceptible to mouse hepatitis virus (MHV) A59 strain infection but resistant to JHM strain, despite the expression of the viral receptor for both strains. This restriction on viral infection has been shown to occur at the virus entry level (Yokomori et al., Virology 196, 45-56, 1993). To study whether JHM resistance of these cell lines is due to a defective cellular factor necessary for JHM virus entry, or due to the presence of an inhibitor, hybrid cells were obtained by polyethylene glycol (PEG)-mediated fusion between two resistant cell lines, i.e., a Balb/c mouse-derived cell line, BC10 cells, which are resistant to JHM infection but express a viral receptor, and the simian cell line COS cells, which are resistant to JHM because of the absence of the viral receptor. JHM could replicate in the hybrid BC10-COS cells, indicating that the restriction of viral infection in BC10 cells could be complemented by COS cells. This result indicates that the resistance of BC10 cells to JHM infection is due to the defectiveness of a cellular factor rather than the presence of an inhibitor. JHM virus-binding and penetration into BC10 cells appeared to be normal. However, JHM internalization into BC10 cells by PEG-induced virus-cell fusion did not lead to viral replication, suggesting that the restriction of JHM infection in BC10 cells is at the level of viral uncoating. This restriction could be complemented by PEG mediated fusion with other murine cell lines which have the virion-uncoating activity, but not by cell lines which lack this activity. Furthermore, the viral resistance of most of other murine cell lines, which express MHV receptors, could not be overcome by fusion with COS cells, suggesting that different murine cell lines are defective in different viral entry functions. Therefore, we conclude that JHM viral entry process requires multiple cellular factors secondary to the viral receptor. PMID- 8249297 TI - Expression of the gene encoding a translational elongation factor 3 homolog of Chlorella virus CVK2. AB - A gene encoding a putative translational elongation factor 3 (EF-3) on the genome of Chlorella virus CVK2 has been cloned and sequenced. It encodes for a predicted polypeptide of 1120 amino acids (aa) with a molecular mass of 127 kDa. The overall amino acid sequence of CVK2 EF-3 (vEF-3) showed 36.1% identity and 83.6% similarity to that of Saccharomyces cerevisiae EF-3. Functional domains including two sets of ATP-binding motifs were extremely well conserved between vEF-3 and yeast EF-3; 63.6% identity and 92.4% similarity in total 330-aa portions. Northern blot analysis indicated that the vEF-3 gene was transcribed in the host cells early, at 20 min postinfection (p.i.), as well as late, 3-4 hr p.i. Western blot analyses with anti-vEF-3 antibody detected the 120 kDa vEF-3 protein product after 40 min p.i. It was present until the final stages of infection but absent in the virion. The vEF-3 gene was highly conserved among all Chlorella viruses isolated in Japan. PMID- 8249298 TI - Transient HTLV-I infection of a human glioma cell line following cell-free exposure. AB - The human T-lymphotropic virus type I (HTLV-I) has been recently associated with cases of tropical spastic paraparesis and human myelopathy. In order to study whether cells of neuroectodermic origin were susceptible to HTLV-I infection, a human glioma cell line T67 was exposed in vitro to HTLV-I by a cell-free method of virus transmission. The presence of HTLV-I proviral DNA was analyzed by polymerase chain reaction 3, 7, and 14 days after infection. The results showed the presence of LTR, pol, and tax sequences within glioma cell line 3 days after the infection. However after 7 and 14 days, detection of HTLV-I sequences remarkably decreased. P19 expression peaked 7 days after infection and decreased in the following week. These data provide evidence that cell-free transmission of HTLV-I results in transient infection of cells of glial origin. PMID- 8249299 TI - NF kappa B upstream regulatory sequences of the HIV-1 LTR are involved in the inhibition of HIV-1 promoter activity by the NS proteins of autonomous parvoviruses H-1 and MVMp. AB - To investigate parvoviral interference with human immunodeficiency virus type 1 (HIV-1) in human cells that are normally susceptible to HIV-1 infection, nonstructural (NS) proteins of the parvoviruses H-1 virus and minute virus of mice were studied for their effect on the activity of the HIV-1 promoter in a variety of CD4+ cells. Transient cotransfection assays revealed a reduced HIV-1 promoter activity in the presence of parvoviral NS proteins. Stimulation of the HIV-1 promoter by phorbol 12-myristate 13-acetate (PMA) led to an increase in its sensitivity to NS-induced suppression. The inhibitory effect of NS polypeptides depended, at least in part, on the presence of the NF kappa B motifs of the HIV-1 long terminal repeat, suggesting an interaction of the parvoviral products with PMA-inducible cellular factors binding to these elements of the HIV-1 promoter. PMID- 8249300 TI - Asp-->Asn substitutions in the putative calcium-binding site of the turnip crinkle virus coat protein affect virus movement in plants. AB - Many plant virus coat proteins have binding sites for divalent cations, particularly calcium. It has been speculated that the purpose of such sites is to ensure that plant viruses release their RNA only in the host cytoplasm which has a low calcium concentration. By comparison to tomato bushy stunt virus, two of the amino acids that probably interact directly with calcium ions in turnip crinkle virus (TCV) capsids, Asp155 and Asp157, were substituted with Asn residues by oligonucleotide-directed mutagenesis to create the mutant TCV-M18. TCV-M18 coat protein, and virions accumulated to wild-type levels in isolated protoplasts. Mutant virions were able to uncoat and to initiate de novo replication in protoplasts although no symptoms were observed in plants inoculated with TCV-M18. Mutant RNA accumulated to much lower levels than wild type RNA in inoculated leaves and was not detected in upper, uninoculated leaves. The lower infectivity of TCV-M18 in plants may be due to a decreased capacity to move efficiently from cell to cell, and we suggest the possibility that TCV coat protein plays an active, pivotal role in cell-to-cell movement interactions. PMID- 8249301 TI - Transcriptional control of the equine herpesvirus 1 immediate early gene. AB - Transient expression assays measuring induction of an equine herpesvirus 1 (EHV 1) immediate early (IE) promoter construct, pIE beta gal, were used to examine trans-induction of the IE gene (IR1) promoter by superinfection with intact EHV-1 (Kentucky A strain), uv-inactivated EHV-1, or EHV-1 stocks highly enriched for defective interfering particles (DIPs), and by cotransfection with plasmids containing EHV-1 DNA fragments. Our results confirm reports by others in that the IE promoter can be induced by both intact and uv-inactivated EHV-1 and provide evidence that DIP-rich virus stocks also possess transinducing activity. Furthermore, IE promoter activity was induced by cotransfection of the promoter construct with either of two plasmids containing restriction enzyme DNA fragments that span ORF12, the putative EHV-1 homolog of the herpes simplex virus 1 (HSV-1) alpha-trans-inducing factor (alpha-TIF) gene, UL48, or by cotransfection with an isolated DNA fragment containing only ORF12, while no transactivation was associated with plasmids linearized by restriction enzyme digestion at a single site within ORF12. These data indicate that, despite its predicted lack of an acidic carboxy terminus (a region essential in HSV-1 alpha-TIF for trans-inducing activity), the EHV-1 ORF12 product is a functional alpha-TIF. PMID- 8249302 TI - DNA-sequence rearrangement required for the adaptation of JC polyomavirus to growth in a human neuroblastoma cell line (IMR-32). AB - Infection of a human neuroblastoma cell line (IMR-32) with the JC polyomavirus (JCV) strain Mad-1 with subsequent serial passage results in the generation of a virus adapted to growth in IMR-32 (K. Akatani, M. Imai, M. Kimura, K. Nagashima, and N. Ikegami, J. Med. Virol., in press). To understand the basis of this adaptation, we molecularly cloned JCV DNAs from the adapted virus. The cloned JCV DNAs consisted of essentially three species (M1-IMRa, -IMRb, and -IMRc) with rearranged regulatory regions. Two TATA sequences are present in the regulatory region of the parental virus Mad-1, but one distal from the origin of replication was commonly deleted in M1-IMRa, -IMRb, and -IMRc. We showed that these regulatory regions were required for the efficient growth of JCV in IMR-32. Various JCV strains should be propagated in IMR-32, if their regulatory regions are replaced with those defined in this study. Since it is difficult to propagate JCV in cells other than primary human fetal glial cells, this system may be useful for structural and immunological studies of JCV. PMID- 8249303 TI - The influence of viral coding sequences on the efficiency of internal initiation of translation of cardiovirus RNAs. AB - Since internal initiation of translation of cardiovirus RNAs requires the approximately 450-nt segment of the viral genome immediately upstream of the authentic initiation codon for viral polyprotein synthesis, the question arises as to whether the immediately adjacent sequences, the start of the polyprotein coding region, also influence the efficiency of internal initiation. Therefore, a variety of constructs derived from encephalomyocarditis virus and Theiler's murine encephalomyelitis virus retaining various lengths of viral coding sequence were translated in rabbit reticulocyte lysates. Efficient internal initiation showed no requirement specifically for viral coding sequences, but on the other hand certain motifs, notably G-rich sequences, located immediately downstream of the initiation codon were highly inhibitory. These results suggest a possible explanation for the evolution of cardiovirus polyproteins lacking N-terminal myristylation signals and are also pertinent to the design of constructs in which the cardiovirus internal ribosome entry signal is used to drive the expression of a reporter cistron. PMID- 8249304 TI - The maize stripe virus major noncapsid protein messenger RNA transcripts contain heterogeneous leader sequences at their 5' termini. AB - Primer extension analyses and a PCR-based cloning strategy were used to identify and characterize 5' nucleotide sequences on the maize stripe virus (MStV) RNA4 mRNA transcripts encoding the major noncapsid protein (NCP). Direct RNA sequence analysis by primer extension showed that the NCP mRNA transcripts had 10-15 nucleotides beyond the 5' terminus of the MStV RNA4 nucleotide sequence. MStV genomic RNAs isolated from ribonucleoprotein particles (RNPs) lacked the additional 5' nucleotides. cDNA clones representing the 5' region of the mRNA transcripts were constructed, and the nucleotide sequences of the 5' regions were determined for 16 clones. Each was found to have a distinct 10-15 nucleotide sequence immediately 5' of the MStV RNA4 sequence. Eleven of 16 clones had the correct MStV RNA4 5' nucleotide sequence, while five showed minor variations at or near the 5' most MStV RNA4 nucleotide. These characteristics show strong similarities to other viral mRNA transcripts which are synthesized by cap snatching. PMID- 8249305 TI - Genetic variation in VP7 gene of human rotavirus serotype 1 (G1 type) isolated in Japan and China. AB - Sequence analysis of the gene encoding the major neutralization glycoprotein (VP7) was performed on 12 human isolates of serotype 1 of rotavirus in Japan and China. They were examined for genetic variations among serotype 1 isolates. Comparative studies of their nucleotide and deduced amino acid sequences between the 12 isolates and the Wa strain revealed an overall homology of more than 92 and 96%, respectively. Higher degrees of homologies were observed between Wa and 2 strains (K1 and K2) in Tokyo, 1979-1980, than between Wa and recent isolated strains in Tokyo and in China. In our isolates, a total of 16 amino acid residues frequently converted to another amino acid. Six amino acid residues belonging to the major neutralizing epitope regions (B, D, and E in this communication) frequently converted. From these data three subtypes (subtypes A, B, and intermediate) were suggested to be divided. Whether these differences are an important mechanism in the epidemiology of rotaviruses requires further investigation. PMID- 8249306 TI - Identification of HIV1 determinants for T lymphoid cell line infection. AB - Determinants responsible for HIV-1 infection of T lymphoid cell lines were identified by functional analysis of chimeric proviral clones derived from T-cell line tropic-(HXB2) and non-T-cell line-tropic isolates (YU2, ADA). Replacement of the HXB2 V3 envelope loop sequence with that derived from YU2 resulted in a virus that is no longer T cell line-tropic. However, the reciprocal replacement using HXB2 V3 loop sequences did not confer upon either ADA or YU2 envelope proteins the ability to infect T cell lines. Furthermore, the resultant viruses were incapable of infection of primary lymphocytes. Single, double, and multiple point mutations made within the V3 loop sequence did not result in change in tropism, although mutations involving residue 275 resulted in a virus that was incapable of infecting primary lymphocytes but retained the ability to infect Jurkat T lymphoid cells. These results suggest that the V3 envelope determinant is necessary for T cell line infection, but other determinant(s) in envelope are also necessary to obtain infectious virus expression. PMID- 8249307 TI - Expansion of the retinal pigment epithelium in experimental myopia. AB - Expansion of the retinal pigment epithelium was studied in neonatal chicks after one or two weeks of unilateral form vision deprivation to investigate altered ocular growth mechanisms in this experimental model of myopia. The area of individual retinal pigment epithelial (RPE) cells, measured in tangential sections, was greater in myopic eyes than in contralateral control eyes at both times. The mean RPE cell area in myopic eyes increased to the same extent as the area of the retinal pigment epithelium as a whole. In control eyes between one and two weeks, RPE cell expansion occurred predominantly in the periphery; in myopic eyes, it occurred more generally across the epithelium but was less pronounced in the temporal region. Given the absence of detectable mitotic figures in control and myopic eyes, expansion of the epithelial layer is attributable to passive stretch or growth of existing cells. Whether scleral growth or stretch occurs selectively beneath the areas of more pronounced RPE cell expansion is unknown. PMID- 8249308 TI - Photochemically-induced lesion of the rat retina: a quantitative model for the evaluation of ischemia-induced retinal damage. AB - The effects of ischemia-induced retinal damage were quantitatively evaluated in rats with the aim of obtaining a suitable model to study the pathogenesis of the loss of retinal neurons after ischemic episodes. Anaesthetized rats were injected with 80 mg/kg i.v. of the fluorescein rose bengal dye and one eye was exposed to cold light for different periods (from 5 to 30 min). The animals were sacrificed at different times (1 and 4 hr; 2 and 7 days) after the lesion and the photochemically-induced damage was evaluated. The damaged retinae appeared thicker, numerous neurons of the inner nuclear layers showed swelling of the perinuclear cytoplasm and the retinal vessels were enlarged. The activity of choline acetyltransferase (ChAT) and glutamic acid decarboxylase (GAD), two marker enzymes of the GABAergic and cholinergic neurons, significantly decreased, indicating a degeneration of GABAergic and cholinergic amacrine cells. PMID- 8249309 TI - Spatial-frequency- and orientation-selectivity of simple and complex channels in region segregation. AB - Models incorporating spatial-frequency- and orientation-selective channels explain many texture-segregation results, particularly when known nonlinearities are included. One such nonlinearity is complex channels. A complex channel consists of two stages of linear filtering separated by a rectification-type nonlinearity. Here we investigate the spatial-frequency- and orientation selectivity of simple (linear) channels and of the complex channels' first stage. Observers rated the degree of segregation between two "textures" both composed of elements which were Gabor patches. When the textures differed in type of element (e.g. one composed of vertical and the other of horizontal Gabor patches), the segregation results yield bandwidth estimates for simple channels of approx. 0.5 1.0 octave on the spatial-frequency dimension and 5-20 deg of rotation on the orientation dimension. When the textures differ in the arrangement of elements (e.g. striped vs checkerboard arrangements, both of horizontal and vertical patches), the segregation results yield bandwidth estimates for the first stage of complex channels. These estimates, while differing substantially from one observer to another, were always substantially wider than those for simple channels (by at least a factor of two) but narrower than bandwidths of LGN cells (particularly on the orientation dimension where LGN cells show little selectivity at all). PMID- 8249310 TI - Linear and rotation motion aftereffects as a function of inspection duration. AB - Subjects rated the strength of linear and rotary motion aftereffects (MAEs) on an eleven point scale. Inspection durations ranged from 30 to 180 sec in 30 sec steps. In Expt 1, trials using a single inspection duration were spaced at least 22 hr apart to minimize the possibility of interactions between adapting stimuli and long lasting aftereffects. In Expt 2, a counterbalanced sequence of inspection durations was completed in a single session. Total duration of the MAE and durations of the decay phase and tail were measured directly. The decay time constant (DTC), the time it takes for rated strength of the MAE to drop to 1/e of its initial value, was calculated from a line fit to a semilog plot of the ratings during the decay phase. The DTC is inversely related to the decay rate which is indexed by the slope of this line. In both experiments, the duration and DTC increased, and decay rate decreased, with increasing inspection duration for both rotary and linear MAEs. This finding replicates the results for linear MAEs and extends them to rotation MAEs. There were no discernible differences between the two types of MAEs. When the trials were spaced, the total duration increased with the square root of inspection duration. The DTC did not follow the square root rule over the entire range but appeared to approximate it for inspection durations of 90 sec and above. When trials were massed, the square root rule did not appear to apply at all. PMID- 8249311 TI - Perceived speed of moving lines depends on orientation, length, speed and luminance. AB - In this study, the perceived speed of a tilted line translating horizontally (for a duration of 167 msec) is evaluated with respect to a vertical line undergoing the same translation. Perceived speed of the oblique line is shown to be underestimated when compared to the vertical line. This bias increases: (1) when the line is further tilted, (2) with greater line lengths, (3) with lower contrasts, and finally (4) with a speed of 2.1 deg/sec as compared to a higher speed of 4.2 deg/sec. These results may be accounted for by considering that two velocity signals are used by the visual system to estimate the speed of the line: the translation of this line (this signal does not depend on the line's orientation) and the motion component normal to the line (this signal depends on orientation). We suggest that these two signals are encoded by different types of units and that the translation signal is specifically extracted at the line endings. We further suggest that these signals are integrated by a weighted average process according to their perceptual salience. Other interpretations are considered at the light of current models dealing with the two-dimensional integration of different velocity signals. PMID- 8249312 TI - The role of features in preattentive vision: comparison of orientation, motion and color cues. AB - Arrays of lines or blobs were used to investigate the role of features vs feature contrast in preattentive vision. This study continues earlier work on orientation cues and extends it into the dimensions of motion and color. Tests were performed on patterns displaying continuous feature gradients, i.e. continuous variation, from element to element, in either line orientation, direction of motion, or color. In different series of experiments, the following four aspects of visual perception were investigated: (i) detection of a salient target ("odd man out" paradigm), (ii) segmentation of texture fields, (iii) search strategies for given targets, and (iv) figure-ground discrimination by grouping. Features were, in general, not found to play an important role in these tasks and performance was instead related to feature contrast. Only in the case of color did performance also depend upon hue, i.e. feature properties themselves. Whereas in all dimensions tested pop-out ("saliency") and segmentation were obtained from target or border elements whose local feature contrast was well above the level of variation elsewhere in the pattern, performance in the two latter tasks differed between orientation, motion, and color. In search, orientation targets were detected quickly when sufficiently distinct from their neighbors, but were apparently searched for serially if feature contrast was similar to that of other elements nearby. Color targets, however, were always detected fast in these patterns, independently of local feature contrast. Also, the perceived grouping of orientation or motion defined targets depended only on local feature contrast and not on the similarity of target elements. In fact, figures of dissimilar elements were seen as easily as figures of similar elements, indicating that, in these dimensions, stimulus coherence is not essential for the discrimination of figure and ground. For color, however, figures made up of the same targets were always seen slightly better than figures composed of different colors. PMID- 8249313 TI - Depth in anticorrelated stereograms: effects of spatial density and interocular delay. AB - Disparity-based depth is not perceived in densely textured, anticorrelated random dot stereograms (RDSs) whose elements carry opposite signs of brightness contrast on corresponding loci, as extant data show. We observed global depth in anticorrelated RDSs flashed repetitively with an interocular delay. During the delay time, a dot array in one eye was paired with a gray frame in the other eye and thus could interact with the negative afterimage of the contralateral dot array. A correlated RDS (e.g. 8 min arc dots, 50% density, 15-msec flash duration) lost depth with delays > 45 msec. An anticorrelated RDS, that was otherwise identical, showed robust depth when flashed with an interocular delay of some 60 msec. A delay was not always necessary to produce depth. At low dot density (1-2%), anticorrelated RDSs showed disparity-dependent local depth even when displayed continuously, or flashed simultaneously; as dot density alone was increased, depth was progressively lost. To make global depth visible in a dense RDS flashed with an interocular delay, the internal response had to be strongly biphasic. Our results support the generally held notion that cyclopean depth signals emerge exclusively from same-sign binocular cortical filters. However, the exclusionary rule may be invalid with respect to the processing of coarse local depth with figural stimuli. Relative depth between a pair of small dots was easily perceived when one of the dots was in opposite contrast, but the depth threshold was then about 0.5 log unit higher than with the same-contrast pair of dots indicating that the internal effects of contrast have not all lost their sign prior to binocular disparity processing. It remains to be determined whether depth can be perceived from edges of opposite contrast. PMID- 8249314 TI - Dichoptic activation of the early motion system. AB - The short range or early motion system has long been considered incapable of binocular integration. We have developed dichoptic motion stimuli which are based upon the decomposition of traveling sinewave gratings into the sum of two standing waves in spatial and temporal quadrature. The monocular views of such displays appear as counterphase flicker but when presented dichoptically the perception is of movement in a unique direction. Two lines of evidence are presented for the binocularity of early motion mechanisms in human vision. First, adaptation to dichoptic motion sinewave gratings is found to result in a motion aftereffect. Second, random texture motion displays based on the quadrature decomposition are found to support dichoptic perception of motion direction, but not figure/ground. Unlike random dot kinematograms, these displays do not necessitate alternating the direction of motion during dichoptic presentation. This encumbrance, and the reliance on figure/ground discrimination, may have been responsible for prior failure to achieve dichoptic motion perception with short range stimuli. PMID- 8249315 TI - Responses of macaque ganglion cells and human observers to compound periodic waveforms. AB - We measured responses of macaque retinal ganglion cells to different periodic waveforms (sinusoidal, square, rapid-on and rapid-off sawtooth waveforms) for both luminance and equiluminant chromatic modulation. We analyzed the responses with a peak-to-trough detector. At low frequencies, on-center and off-center magnocellular (MC-) pathway cells showed a ten-fold higher responsivity to the rapid-on and rapid-off sawtooth respectively. Red-on (+L-M) and green-on (+M-L) parvocellular (PC-) pathway cells showed a four-fold greater responsivity to rapid red-on and rapid green-on equiluminant chromatic sawtooth waveforms respectively. At an equivalent retinal eccentricity, we measured psychophysical thresholds for luminance stimuli and chromatic stimuli. We concluded that luminance sawtooth sensitivities from psychophysics are consistent with selective detection through MC-pathway on- and off-center channels in the visual system. The differences between the compound periodic waveforms seen in the PC-pathway cell data did not occur in the psychophysics. In a second analysis, cell responses to sinusoidal modulation were used to predict the linear response to square-wave and sawtooth waveforms. PC-pathway cells showed linear temporal behavior over a wide range of contrasts, but MC-pathway cells displayed linear behavior only for low-contrast luminance modulation. Using these linear fits, we implemented a model incorporating central low-pass filtering in the MC- and PC pathways before the peak-to-trough detector. This model captured better the time scale and relative sensitivity to periodic waveforms found in the psychophysical data. PMID- 8249316 TI - Dependence on stimulus onset asynchrony in apparent motion: evidence for two mechanisms. AB - The detection of the direction of motion was measured as a function of the spatial and temporal offset for a kinematogram stimulus presented in two-frame apparent motion. The stimulus was made up of Gabor function micro-patterns randomly distributed across the stimulus field. We show that for short stimulus onset asynchronies (SOA) performance can be predicted from the spatio-temporal Fourier power spectrum of the stimulus, whereas for long SOAs the pattern of performance is qualitatively different from such a prediction. The dependence of motion perception on SOA exhibits an abrupt change from one mode of behaviour to the other. These findings are suggestive of the operation of distinct mechanisms, one "quasi-linear" and one "nonlinear", which can be separated by temporal parameters. PMID- 8249317 TI - Nonlinear combination rules and the perception of visual motion transparency. AB - Experiments were performed to elucidate the decomposition performed by the human visual system in the segregation of complex motion stimuli into distinct moving surfaces. Subjects were presented with achromatic patterns consisting of four types of elements, generated from two random binary luminance patterns (random dot checkerboards). The luminance of each of the four region classes was under program control. Animated sequences of such images were produced by displacing each of the two generating patterns in opposite directions on a frame by frame basis. These displays evoke a wide variety of percepts, depending on the programmed luminance values, including motion in a single direction, simultaneous motions of transparent sheets in opposite directions, dynamic noise with no directional component, or any combination of the above percepts. A theory is presented which relates the strengths of these percepts to the amplitudes of the components in the perceptual decomposition. The experiments described measured thresholds for seeing noise or "twinkling" in addition to the multiple motions, with the goal of determining the particular signal transformations preceding motion analysis. The results are consistent with a motion extraction mechanism which operates on a linear representation of the input imagery. These results extend a similar finding due to Anstis and Mather [(1985) Perception, 14, 167 179] and call into question the interpretation of a recent study by Stoner, Allbright and Ramachandran [(1990) Nature (London), 344, 153-155]. PMID- 8249318 TI - Interactions of spectral, spatial, and temporal mechanisms in the human pattern visual evoked potential. AB - The human pattern onset-offset visual evoked potential (VEP) was studied with different colours and spatial frequencies presented on a steady homogeneous intensive yellow background. Under this condition a broad, late negative wave (N2) dominated by the blue-sensitive mechanism and a sharper earlier positive wave (P1) dominated by the red-green-sensitive mechanism can be observed. With a 460 nm pattern N2 shows a strong amplitude tuning at low-medium spatial frequencies. With 550 nm an early negative component (N1) is added showing an amplitude tuning at high spatial frequencies. Different spatial resolutions of the two colour mechanisms are thus indicated. With pattern-reversal stimuli the 550 nm stimulus shows a higher temporal resolution than the 460 nm stimulus. PMID- 8249319 TI - Functional brain imaging: dipole localization and Laplacian methods. AB - The performance of two methods, used to localize brain activity from evoked potential fields measured on the scalp, was assessed in a tank model of the human head. This physical model contained a human skull encased in a polymer simulating the resistivity and geometry of brain and scalp. The dipole localization method mislocalized the positions of known dipole sources by several centimeters. The mislocalization was systematic. The dipoles were localized too deeply in the head. The Laplacian method yielded a field resembling the brain surface field (epicortical potential field) provided that the iso-potential contours of the scalp field closed within the measurement range. Clipping resulted in a serious mislocalization of the position of the peak of the epicortical potential field. PMID- 8249320 TI - Light-induced binding of proteins to rhabdomeric membranes in the retina of crayfish (Procambarus clarkii). AB - Light-induced protein interaction as part of the process of visual transduction in arthropods with rhabdomeric photoreceptors was investigated biochemically by using crayfish retina. Two kinds of retinal buffer soluble proteins (one of 40 kDa and the other of 46 kDa) were found to bind to the irradiated rhabdomeric membranes both in vitro and in vivo. The proteins bound to the membranes in the presence of metarhodopsin. An antibody against mouse arrestin (S-antigen) cross reacted with the 40 kDa protein. These results suggest that the binding of the proteins to the membranes is caused by the formation of metarhodopsin, and that the 40 kDa protein has a similar structure to arrestin. PMID- 8249321 TI - Strain differences in the ratio of synaptic body types in photoreceptors of the rat retina. AB - In the retinal outer plexiform layer of seven different rat strains, synaptic bodies (SB) were counted and, according to their morphology, characterized as synaptic ribbons (SR), synaptic spheres (SS) or intermediate structures. It was found that absolute SB numbers showed relatively small variations while SR/SS ratios differed considerably between the strains investigated. These results are discussed with respect to retinal pigmentation and to formation and degradation, respectively, of synaptic ribbons. PMID- 8249322 TI - Comparison of the waveforms of the ON bipolar neuron and the b-wave of the electroretinogram. AB - The b-wave of the electroretinogram (ERG) is believed to result largely from the light responses of ON bipolar neurons. One apparent inconsistency is that the b wave is a phasic signal, more similar to ON responses of transient third-order neurons than to the ON bipolar light responses. However, when selective glutamate agonists or antagonists were used to isolate the b-wave, it was found to be tonic and resemble the light response waveform of the ON bipolar cell. PMID- 8249323 TI - On the possible function of the proximal lens unit of Notonecta glauca. AB - The water bug Notonecta glauca has doublet lenses corrected for spherical aberration. The proximal lens unit constitutes a refractive index transition between the distal lens unit and the crystalline cone. It inevitably reduces the reflectivity of the eye. Such correction for spherical aberration, however, could also be simply achieved by means of an aplanatic singlet lens. Therefore it seems possible to suppose that the proximal lens unit might produce both decreased reflection and increased transmission. To test this idea, the reflectivity of the doublet in Notonecta is calculated and compared with the reflectivity of two corresponding aplanatic singlets of different refractive indices for contact with air and water. Because of the lower lens unit, the absolute reflection suppression delta R in the dioptric apparatus is about 2% in water and the relative reflectivity reduction delta R/R amounts to 65%. In air this effect is negligible. The question of the evolutionary importance of this phenomenon is briefly discussed. PMID- 8249324 TI - Chronotopographical analysis of the pattern onset visual evoked magnetic response (VEMR): implications for waveform peak identification. AB - The topography of the visual evoked magnetic response (VEMR) to a pattern onset stimulus was studied in five normal subjects using a single channel BTi magnetometer. Topographic distributions were analysed at regular intervals following stimulus onset (chronotopography). Two distinct field distributions were observed with half field stimulation: (1) activity corresponding to the C11 m which remains stable for an average of 34 msec and (2) activity corresponding to the C111 m which remains stable for about 50 msec. However, the full field topography of the largest peak within the first 130 msec does not have a predictable latency or topography in different subjects. The data suggest that the appearance of this peak is dependent on the amplitude, latency and duration of the half field C11 m peaks and the efficiency of half field summation. Hence, topographic mapping is essential to correctly identify the C11 m peak in a full field response as waveform morphology, peak latency and polarity are not reliable indicators. PMID- 8249325 TI - Express saccades: the effects of spatial and temporal uncertainty. AB - Once trained to make express saccades, monkeys continue to make express saccades when there is a high level of uncertainty about both the location and the time of onset of a peripheral target. Providing additional information about the location and the time of onset of a peripheral target does not increase the frequency of occurrence of short latency saccades. Furthermore, the metrics of saccades with express latencies are typical of visually-guided saccades, rather than anticipatory movements or saccades to the remembered location of a target. Finally, practice on the gap paradigm restricted to one target location always viewed from the same initial eye position does not increase the probability of producing express saccades to the same target from other initial eye positions. Instead, the practice effect is specific to targets that have the same relative location, regardless of initial eye position. PMID- 8249326 TI - Influence of the contrast sensitivity function on the reaction time. AB - The reaction time (RT) vs spatial frequency (SF) curve is determined, using gratings from 1 to 40 c/deg, at seven different contrast levels between 0.95 and 0.02. The form of the RT/SF function: (a) replicated the inverse of the contrast sensitivity function (CSF) at near threshold contrast levels; (b) behaved differently at higher contrasts, exhibiting two branches at contrast close to 1. The interpretation is that there are two factors determining this function: (1) the transition from the operation of fast transient channels at low SF to the operation of slow sustained channels at high SF, the transition taking place within a narrow SF band close to 6 or 8 c/deg (depending on the subject) and (2) the contrast attenuation by the optical and neural transfer function, operating throughout the SF range. At high contrasts, the effect of the first factor can be clearly observed, because the effect of the second factor does not change with spatial frequency except in a region where the RT/SF function changes rapidly. At lower contrasts, however, the second factor becomes increasingly relevant while the first becomes less and less observable. PMID- 8249327 TI - Mechanisms of velocity constancy. AB - Human observers can compare the physical velocities of objects (cm/sec) moving at different distances quite well, although the objects' retinal velocities (deg/sec) may vary considerably. This perceptual ability is called velocity constancy. We conducted a number of experiments to investigate what mechanisms observers use to attain this constancy and if pure motion signals can also be matched according to their physical speeds. Subjects were asked to match the velocities of two moving stimuli presented at different viewing distances. The stimuli consisted of sparse random-dot kinematograms or drifting Julesz patterns. The subjects matched the true physical velocities of the stimuli provided that the two visual scenes contained identical size references. Knowledge of the actual viewing distances proved to be irrelevant for evaluating the physical velocities of the stimuli. We conclude that velocity constancy is based upon a relative scaling algorithm. PMID- 8249328 TI - Dependence of plaid motion coherence on component grating directions. AB - We measured motion coherence for plaids composed of two different spatial frequency (SF) cosine grating components moving at various relative angles. The component SFs were in a ratio of 6:1, and several component motion directions were chosen to produce small to large angular differences. For angles of less than +/- 45 deg all subjects perceived rigid coherent motion, while for angles of +/- 45 deg or greater the components were perceived to slide transparently. The results were not altered when we changed the ratio of component contrasts or speeds over a factor of 2 and varied the SF ratio up to 9:1. These results show that transparency or coherence of moving plaids in different spatial scales depends on the relative component motion directions and is relatively independent of contrast, speed, and SF difference between the components. This angular dependence also explains recent data previously thought to be based on a visual computation of multiplicative transparency. A quantitative model in which the resultant motion on each scale provides a facilitative biasing signal to units tuned to similar directions (within +/- 30 deg) on other scales explains the experimental results. PMID- 8249329 TI - The effect of contrast polarity on letter identification. AB - To determine the effect of contrast polarity on the spatial characteristics of letter identification, we measured contrast sensitivity for individual Sloan letters that were presented either as luminance increments or luminance decrements relative to a continuously presented adapting field. The temporal mode of presentation consisted of either a rapid onset with a Gaussian offset, or the reverse. When contrast was specified in terms of Rayleigh (Michelson) units, the contrast sensitivity function for letters of positive contrast extended to smaller letter sizes than the function for letters of negative contrast. However, when contrast was defined in Weber terms, letter contrast sensitivity functions were identical for letters of positive and negative contrast, indicating that letter identification was equivalent for luminance increments and decrements that had equal absolute magnitude. Onset-offset characteristics had no differential effect on letter contrast sensitivity by either contrast definition. These findings provide a basis for predicting the effect of contrast polarity on tasks that involve letter identification. PMID- 8249330 TI - The effect of edge blur on grating induction magnitude. AB - In order to assess the contribution of high spatial frequency channels (i.e. local, edge-dependent mechanisms) to the grating induction effect, grating induction magnitude was measured as a function of systematic amounts of blurring of the inducing/test field boundary for four test field heights which spanned a two octave range (0.25-2.0 degrees). Measurements were obtained from two subjects using both the cancelling procedure of McCourt [(1982) Vision Research, 22, 119 134] and a contrast matching paradigm. The two measures yielded similar outcomes: consistent with previous results, both matching and cancelling contrast decreased monotonically with increasing test field height. The effect of blurring the edge was to produce a small (eta 2 = 0.6-5.1%), but significant (P < 0.001), increase in grating induction magnitude. A second experiment utilized the contrast matching paradigm to investigate the effect of edge blur at extreme values (i.e. zero blur and maximum blur) on grating induction magnitude across a four octave range of spatial frequency (0.0625-1.0 c/deg). The results of the matching procedure were again consistent with those obtained previously using the cancelling technique: grating induction magnitude decreased monotonically with increasing spatial frequency. The effect of blurring was to produce a modest (eta 2 = 1.0-2.8%), but significant (P < 0.001), elevation in induction magnitude. These results lead to the conclusion that, unlike some other brightness effects, visual spatial filters selectively sensitive to high spatial frequencies, or to the edges which they sharpen, are not essential for the production of the grating induction effect. PMID- 8249331 TI - Orientation acuity for sine-wave gratings with random variation of spatial frequency. AB - Orientation discrimination thresholds were estimated for sine-wave grating stimuli using a two-alternative forced-choice procedure on the two principal, and the two main oblique axes. The two discriminanda within any trial were subject to independent random variation of their spatial frequency. The variation of spatial frequency was defined by a Gaussian probability density function. Thresholds were obtained for five different depths of frequency variation within the range 0.0 1.0 octave. We find that orientation acuity on any of the axes tested is unaffected by variation in the spatial frequency of the targets. The data support the hypothesis that orientation and spatial frequency are independent and orthogonal neural codes. PMID- 8249332 TI - Motion perception with spatiotemporally matched chromatic and achromatic information reveals a "slow" and a "fast" motion system. AB - Recent reports dealing with apparent motion challenged the standard view according to which motion processing should be impossible if the visual attributes matched across space and time are processed in independent channels (the similarity principle). The present work examines this possibility insofar as it relates to the spatiotemporal combination of pure chromatic and pure luminance information. The data indicate that the "similarity principle" is indeed infringed at low (< or = 2.5 Hz, i.e. velocities of 2.5 deg/sec for spatial modulations of 1 c/deg, in this study) but not at high (> or = 7.5 Hz) temporal frequencies. The fact that colour and luminance may or may not combine to yield motion perception depending on their temporal modulation reconciliates contradictory results in the literature and supports the idea of two motion systems, a "fast"/specific one, integrating information only from similar subunits, and a "slow"/unspecific one, integrating information across dissimilar subunits (in the present case, across the chromatic and achromatic "domains"). This dichotomy is also supported by the finding that chromatic reverse-phi (i.e. with equiluminant, red and green stimuli) can be observed at medium temporal frequencies but is replaced by direct motion at low temporal frequencies, presumably within the range of the "slow"/unspecific system. Using a modified "minimum motion" technique (referred to as the Reverse-Phi equiluminance method) we present data allowing to assess the relative weights of the two systems as a function of temporal frequency. PMID- 8249333 TI - Development of infant contrast sensitivity to chromatic stimuli. AB - We have monitored the development of contrast sensitivity to equiluminant red green chromatic patterns by monitoring visual evoked potentials (VEPs) in 13 infants. The results confirm our previous report [Morrone, Burr and Fiorentini, Proceedings of the Royal Society B, 242 (1990a)] that, before 7-8 weeks of age, there was no response to purely chromatic stimuli, while at the same age luminance stimuli of 20% contrast produced reliable responses. At all ages (even before the onset of a chromatic response) the colour mixture to yield equiluminance was similar to that of adults, suggesting that the relative proportion and efficacy of medium- and long-wave cones is similar for infants as for adults. For both luminance and chromatic stimuli, amplitude increased roughly linearly with log-contrast, so sensitivity thresholds could be predicted by linear extrapolation to the abscissa. Detailed contrast sensitivity curves were measured for four infants at various ages. The results show that luminance and chromatic contrast sensitivity develop independently at different rates, probably reflecting differential development of postreceptoral neural mechanisms. PMID- 8249334 TI - Does chromatic sensitivity develop more slowly than luminance sensitivity? AB - Chromatic sensitivity is very low in humans during the first few months of life. We examined whether low chromatic sensitivity reflects a deficiency among chromatic mechanisms or whether it is simply a manifestation of poor visual sensitivity in general. The sweep VEP was used to measure contrast sensitivity to gratings varying in the mixture of red and green components. For infants from 2 to 8 weeks of age, sensitivity to all mixtures was lower than color-normal adults' sensitivity, but infant and adult ratios of luminance/chromatic sensitivity were similar. This finding is consistent with the hypothesis that infants have functional MWS and LWS cones and the requisite post-receptor chromatic mechanisms to compare their signals. PMID- 8249335 TI - Multi-aperture viewing: perception of figures through very small apertures. AB - A new phenomenon of viewing objects through multiple apertures is reported. When a solid figure is hidden behind a sieve of very small apertures (pinholes), only its general shape may be perceived. Outline figures may be entirely invisible behind the sieve. Motion of the figure markedly improves the visibility of the outline figures and of the small details and edges of the solid figures. Two experiments are presented. The first demonstrates that the size of a dark stimulus moving behind a rear-illuminated row of holes is perceived with an accuracy that may be considerably better (i.e. the error is smaller) than the interhole distance. The results of the second experiment show that the visibility of an outline figure, a ring, is significantly better when the figure smoothly moves than when presented at random positions behind a two-dimensional sieve. Changing the velocity within the range of 5.18-10.36 deg/sec does not affect the visibility of the figure while it moves; however, a lower rate of discrete presentation leads to significantly better visibility. An explanation of the phenomenon is presented in terms of integration of information about the relative positions of the covered/uncovered holes, with a possible involvement of motion analyzing mechanisms. PMID- 8249336 TI - Preattentive discrimination of relative phase modelled by interacting Gabor or by difference-of-Gaussian filters. AB - A computer simulation of the human preattentive visual pathway used Gabor and difference-of-Gaussian (DOG) filters to model two-dimensional relative phase discrimination. There is a hierarchy of hexagonally packed levels: (i) an image layer; (ii) an intermediate level of on- and off-centre DOG filters; (iii) a partial set of broadband oriented Gabor-like filters with high-level DOG filters in parallel. Connections between layers use half-wave rectification and a compressive nonlinearity. Local feedback interactions between oriented Gabor filters, together with spatial averaging, allow the model to discriminate both two-dimensional relative phase and orientation differences. There were two separate simulations for the Gabor filters, one with even-symmetric filters and another with odd-symmetric. Textures were formed by superposing three high contrast sine-wave gratings with successive rotations of 60 degrees. Relative phase and global orientation were the varied parameters. Psychophysical rating data for peripheral viewing of texture pairs resemble the results from the even symmetric simulation, showing discrimination of relative phase and orientation. In contrast, the DOG filters in the model simulate only the relative phase aspects of the data. PMID- 8249337 TI - [The organization of the treatment of minor casualties in neurosurgery]. AB - The selection of neurosurgical minor wounded during casualty staging has a number of specific features. First of all it is caused by objective difficulties in the diagnosis of wounds and traumas of nervous system. Patients who have even minimal traumas of soft tissues of head or bone structures of skull could have intracranial injuries which could be treated only at the stage of tertiary medical care. These patients can be evacuated to the hospital for minor wounded if they have no confirmation on intracranial injuries. PMID- 8249338 TI - [Current problems in organizing medical care for those with minor closed brain trauma]. AB - The approach to the management of slight nonpenetrating cerebral casualties must be seriously revised in order to repudiate the idea to consider these traumas as "the last residual element". Multiple defects in diagnosis and treatment of such trauma interconnect with various posttraumatic syndromes, which can be disclosed by close examination. Frequently the postcontusional disorders occur in this category of wounded because of their classification as "minor casualties". The prevention of scar and commissure processes, atrophies, hypertensive or hydrocephalic disorders, as well as diminishing of incapacitation rate could be achieved by improved methods of treatment, opportune application of modern diagnostical facilities, and broad clinical experience gained in field conditions. Nevertheless, many organizational problems of diagnosis and treatment have no clear-cut decisions and need further scientific researches. PMID- 8249339 TI - [The organization and principles of rendering medical care to minor casualties in otorhinolaryngology]. AB - The article deals with the notions of prior and coexistent ENT injuries. There are classification of mechanical injuries and ENT diseases, as well as rate and structure of ENT sanitary losses, and its evacuation and treatment procedures. The authors give gradations of slight, medium and severe ENT-injured and ENT-sick contingent, and set up the categories of minimally ENT sick, injured and wounded. The article also contains data concerning the treatment policy including immediate medical care at various stages of medical evacuation, and describes peculiarities of triage of different groups of ENT sick and injured. There are proposals for improvement of medical care to ENT sick, injured and wounded. PMID- 8249340 TI - [The prospects for improving medical care for minor casualties and patients in ophthalmology]. AB - As for modern war the casualty rate of ophthalmological minor wounded will be approximately 1.7% of all sanitary losses. The improvement of medical care to this category of patients will be possible if some organizational principles are observed: selection of these patients into a separate group in order to promote their rapid evacuation to the hospital for minor wounded (HMW); existence of physician-ophthalmologist at the stage of secondary care for qualified triage, immediate and holding care, and pre-evacuation procedures; application of modern ophthalmological facilities in field conditions; optic correction of sight in patients at HMW. PMID- 8249341 TI - [Anesthesiological care in the treatment system for minor casualties]. PMID- 8249342 TI - [The nature and characteristics of treating nonpenetrating abdominal wounds]. AB - According to the analysis of 2687 medical histories of abdominal injuries during the 9 years of the Afghan war 25% of them were nonpenetrating wounds. The basic surgical intervention in these cases was a primary surgical treatment of wounds. Laparotomy was performed for 10% of patients because of suspicion of internal injuries. After admission of such patients to medical establishments they must pass a thorough examination. PMID- 8249343 TI - [The therapeutic aspects of the treatment and rehabilitation of minor casualties in neurosurgery]. AB - The article is based on the data obtained in the result of examination of 53 wounded with closed cranial injuries (slight and middle gravity cases), and 34 wounded with spine injuries, but without spinal marrow traumas. The activity of fibroblasts of spinal marrow was studied, as well as the functional activity of peripheral blood monocytes during a ten-day posttraumatic period. It was found out that patients had refractory anemia with a diminishing number of erythroid cells, and also initial infectious complications, such as pneumonia or wound suppuration. PMID- 8249344 TI - [The use of therapeutic soft contact lenses for eliminating blepharospastic blindness in eye burns of mild to moderate degrees of severity]. AB - Application of soft contact lens made of hi-hydrofiled polymeric material for patients with blepharospastic blindness makes it possible to remove or considerably diminish the corneal syndrome in slight and middle gravity burns. Most patients could be returned to their ranks and occupations. PMID- 8249345 TI - [Problems in the early rehabilitation of minor casualties with injuries to the ENT organs]. AB - The article examines the questions of a specialized otorhinolaryngological care to the minor wounded with mine injuries. There are description of evacuation procedures for this category of wounded. Up to 70% of such patients can be treated at the HMW. The article contains schemes concerning the conservative treatment of auditory disorders caused by mine traumas. The authors describe the methods of conduction anesthesia via the low palpebral fissure in cases when patients have traumas of the middle facial zone. PMID- 8249346 TI - [Medical care delivery and specialized treatment for patients with minor dermatovenereal disease]. PMID- 8249347 TI - [The organizational aspects of treating light casualties in modern warfare (2)]. AB - The experience gathered by Medical Service during the war in Afghanistan and during liquidation of the consequences of various disasters and accidents has shown that the most rational method of treatment of minor wounded near the combat area or zone of disaster was a two-staged (and sometimes a three-staged) management system. At the combat tactical zone it is expedient to render the secondary surgical care and reanimation procedures of vital cases, and also provide treatment of minor wounded who could be returned to their ranks in 10 days. For this purpose it is necessary to integrate the sections of medical triage and minor wounded treatment into organic structure of the Brigade medical company and Divisional hospital. As for Army Medical Brigade it must have in its structure a hospital for minor wounded who could be returned to their ranks in 20 days. All the wounded who have to be treated more than 20 days must be evacuated to the Front hospital for minor wounded. PMID- 8249348 TI - [The characteristics of providing an epidemic-control regimen in military field hospitals for minor casualties]. AB - On the basis of the experience gained during the Great Patriotic war (1941-1945) on antiepidemic support of minimally wounded, and analysing the work of the military field hospitals for minimally wounded (HMW) the authors describe the role played by the "mixing" factor and the "local population transmission" factor upon the spreading of infectious diseases. A special attention is paid to disclosure of infectious patients during their admission to the HMW, measures towards the enhancement of their immune resistance, and strict control over sterilization of medical instruments. A proposal was made to introduce a post of sanitation physician in the organic structure of HMW. PMID- 8249349 TI - [The organization of medical care for patients with minor infectious pathology]. AB - Taking into account the prognosticated modern character of combat actions, a new organic structure of Russian Armed Forces, and estimated structure of infectious diseases in war period, it was stressed that anti-infectious platoons (inside medical companies of Brigades) would perform the main part of diagnosis and treatment of minimally sick patients (85% from all infectious cases). There are proposals towards elaboration of a new organic structure of the military field infectious hospital. PMID- 8249350 TI - [The hygienic characteristics of the working conditions for marines during a landing operation]. AB - The article discusses the questions of the marines' activities during the mounting and staging of naval assault operation. The authors describe unfavourable factors of environmental conditions and limitations of manned objects of military technique. There are data concerning the health status and operational ability of the marines depending on characteristic features of various climatic zones, and stages of naval assault operation. The article emphasizes the role of sanitary-epidemiological control over water supplies, meals, and accommodation of servicemen in field conditions. PMID- 8249351 TI - [The medical aspects of nutrition in minor casualties]. AB - It is recommended that a standard ration will be accompanied by "Unpit", natural casein concentrates, and "Glucosolan" vitaminized preparation to eliminate++ the disorders of trophological status of minimally wounded. There is a proposal to include the products of enteral nutrition for the ration of food supplies, because it would shorten the recovery time of minimally wounded, and, thus, accelerate their return to the ranks. PMID- 8249352 TI - [The principles of organizing medical care for the minor casualties of Air Force units under battle conditions]. AB - The article describes methodological aspects of medical care to minor wounded of the Air Force servicemen in the conditions of modern war. There are data concerning the scale and structure of sanitary losses among the minimally wounded of an Air Army, as well as proposals concerning the optimal utilization of the military field air force hospitals in the conditions of the modern war front defence operation. PMID- 8249353 TI - [The significance today of the experience in treating minor casualties in the Navy during World War II]. PMID- 8249354 TI - [The characteristics of medical supplies in rendering medical care in minor casualties and minor illnesses]. AB - The article describes limitations of the existing system of medical supply. There are proposals towards the improvement of the organic structure of this service, storage and fabrication of medications in field conditions. PMID- 8249355 TI - [The improvement of field materiel and equipment for rendering medical care in minor casualties, their treatment and rehabilitation]. PMID- 8249356 TI - [Medical support for Soviet troops in the Battle of Kursk (on the 50th anniversary of the engagement)]. PMID- 8249357 TI - [The principles of a differential approach to rendering medical care and treatment for light casualties in wartime]. PMID- 8249358 TI - Erythropoietin: physiologic basis for clinical applications. PMID- 8249359 TI - Prenatal management of fetal alloimmune thrombocytopenia. PMID- 8249360 TI - Effectiveness of four intravenous IgG preparations in prevention of phagocytosis of anti-Rh-(D)-coated erythrocytes by mononuclear phagocytes. AB - Four commercial intravenous immunoglobulin G (IGIV) preparations, Sandoglobulin (Sandoz), Intraglobin (Biotest), Gammonativ (Kabivitrum) and Gammaogard (Kabivitrum) showed percent inhibition of Fc receptor-mediated phagocytosis of anti-Rh(D)-coated erythrocytes as follows: 45 +/- 14 SD, 41 +/- 23 SD, 35 +/- 14 SD, 25 +/- 13 SD, respectively. Pre-incubation of Rh(D)-positive and antibody coated Rh(D)-positive erythrocytes with IGIV did not lead to any inhibitory effects on phagocytosis. However, preincubation of mononuclear phagocytes with IGIV gave marked inhibition of phagocytosis of antibody-coated erythrocytes. In summary, all four IGIV preparations showed Fc receptor-mediated inhibition of phagocytosis of anti-Rh(D)-coated erythrocytes by mononuclear phagocytes. PMID- 8249361 TI - Role of lactate in platelet storage lesion. AB - It is known that lactate accumulation may cause a pH fall in platelet concentrates (PC) during storage, and this phenomenon causes platelet morphological lesions and loss of platelet in vivo viability. In this study, we added increasing amounts of lactate to identical PC in order to evaluate the role of hydrogen ion accumulation in determining platelet activation and lesion during storage. Six hours after PC preparation, lactate was added to PC1 and PC2 at 20 and 12 mM final concentrations, respectively, while PC3 served as control. In PC1, pH was lower than 6.3, and platelet function and discoid morphology were lost. PC2 were stored for 7 days at pH values ranging from 6.4 to 6.6, and most results of in vitro measurements reflecting platelet function such as osmotic reversal, ATP release and aggregation in response to different stimuli were not significantly inferior when compared to controls. The addition of lactate had no apparent effect on the rise of platelet activation markers P-Selectin, lysosome like protein gp 53, platelet-bound fibrinogen and granulophysin, while a reduction of borderline significance was observed in glycoprotein Ib expression after pH reduction to values lower than 6.6. It is concluded that the rise of platelet activation markers during storage reflects platelet lesions different from those determined by lactate per se. PMID- 8249362 TI - Prevalence and consistency of ALT elevation in plasmapheresis donors: implications for the assessment of blood product infectivity. AB - Alanine aminotransferase (ALT) levels were determined in 8,420 plasmapheresis donations obtained from 431 donors over a period of 18 months. Using sex differentiated normal ranges 2.5% of donations but 23% of donors exhibited elevated ALT levels on at least 1 occasion. Amongst the donors with elevated ALT this was only seen on 1 occasion in one third, while a quarter had elevations in consecutive donations. No donors with consecutive elevations above 100 IU/l were detected. The results are discussed in terms of the guidelines currently recommended for assessing post-transfusion hepatitis infectivity of blood products, such as factor VIII. It is concluded that the current allowance for infection acquired from sources other than blood products under consideration may be over-generous, leading to a potential underestimate of the true rate of infection. PMID- 8249363 TI - Analysis of pre- and post-donation haematological values in plateletpheresis donors. AB - Pre- and post-donation haematological values were measured in 112 donors undergoing plateletpheresis using Haemonetics PCS machines. We found significant differences between pre- and post-donation means for males (M) and females (F) for haemoglobin (M: pre 14.73 g/dl, post 15.25 g/dl; F: pre 13.57 g/dl, post 13.94 g/dl), haematocrit (M: pre 0.418, post 0.431; F: pre 0.389, post 0.4), total protein (M: pre 73.1 g/l, post 66 g/l; F: pre 72.2 g/l, post 63.7 g/l) and albumin (M: pre 42.2 g/l, post 38.5 g/l; F: 41.4 g/l, post 37 g/l). Significant differences were also seen for platelet count (pre 258.6 x 10(9)/l, post 229.2 x 10(9)/l), total white cells (pre 5.3 x 10(9)/l, post 5.55 x 10(9)/l) and neutrophils (pre 3.15 x 10(9)/l, post 3.33 x 10(9)/l), but there were no differences between males and females. This information was of value in establishing post-donation reference ranges which could be utilised when reviewing the suitability of donors for subsequent donations. PMID- 8249364 TI - Recombinant human erythropoietin as adjuvant treatment for autologous blood donation. A prospective study. AB - In a prospective randomized study we investigated the potential of subcutaneous recombinant human erythropoietin (rhEpo) as adjuvant treatment for autologous blood transfusions (3 units) in elective surgery. Four and 2 weeks before surgery, 49 patients received 6 x 10,000 U of rhEpo. delta Hb values (days -28 and 0) of the rhEpo group were compared to delta Hb values of 52 controls (no rhEpo). Reticulocytes were measured at days -21, -14, -7 and 0. Peri- and postoperative supplementary homologous blood requirements were compared in the two randomized groups. delta Hb of rhEpo group was 0.96 g/dl (mean value) and 2.38 for controls. Reticulocyte count increased earlier and to higher levels in rhEpo-treated patients. Except in 1 case, Epo was well tolerated. These results indicate that autologous predonation (3 x 400 ml) does not create anemia if adjuvant Epo treatment is given. However, homologous blood requirements were not significantly different, which is probably due to the fact that 96 of the 101 treated patients underwent elective orthopedic surgery requiring limited blood replacement. Significant benefit of the Epo regimen can be expected in elective cardiovascular and hepatic surgery where larger amounts of blood (5-6 units) are needed. PMID- 8249365 TI - Evaluation of monoclonal antibodies with specificity for the human platelet HPA-1 allotypes. AB - Two mouse monoclonal antibodies have been characterized. Both antibodies specifically recognized the amino acid sequences which are responsible for the presence of the HPA-1a and HPA-1b alloantigens at the surface of human platelet. The monoclonality allied with an exquisite specificity of these antibodies allow a perfect typing of platelet HPA-1 status. A fast and reliable Elisa assay which fulfills all the needs and requirements of hematology laboratories has been set up and is described here. The Elisa assay has been tested on a population of 689 blood donors. The results agreed with the known phenotypic frequencies of the HPA 1 alloantigens. PMID- 8249366 TI - Influence of chloroquine or acid treatment of human platelets on the antigenicity of HLA and the 'thrombocyte-specific' glycoproteins Ia/IIa, IIb, and IIb/IIIa. AB - The influence of treatment of platelets with citrate buffer (pH 7.2), chloroquine, or citric acid at pH 3 on the expression of HLA class I antigens and 'thrombocyte-specific' glycoproteins was investigated by means of flow cytometry. After treatment with citric acid at pH 3 and chloroquine, the expression of HLA class I was significantly reduced, while the density of the molecules GPIa/IIa, GPIIb, and GPIIb/IIIa (GP = glycoprotein) carrying 'thrombocyte-specific' antigens was not or only weakly decreased on the surface of the platelets. The use of two monoclonal antibodies (HC-10 and HC-A2) against the native heavy chain of the HLA class I molecule revealed that 'antigen stripping' with chloroquine or citric acid does not affect the entire molecule: only the beta 2-microglobulin is cleaved, or only some epitopes on the heavy chain are altered by this procedure. The treatment with citric acid yielded better results with respect to the removal of HLA class I activity and the preservation of 'thrombocyte-specific' glycoproteins. The presence of the heavy chain of HLA class I molecules on the surface of platelets after treatment with citric acid and chloroquine confirms the hypothesis that platelets--like nucleated cells--bear HLA class I antigens inserted in the cell by a cytoplasmic and a transmembrane domain. PMID- 8249367 TI - Use of jacalin as a solid phase in ABO reverse grouping. AB - A major problem of using red cells as the solid phase in assay systems is the difficulty to bind them strongly to appropriate surfaces. We report here on a number of lectins of different specificities which were examined for their ability to bind red cells to polystyrene 96-well microtitre plates. The use of the Thomsen-Friedenreich antigen-specific lectins, jacalin, mushroom and Maclura pomifera agglutinin proved the most useful for ABO reverse grouping. Jacalin coated plates were also compared with plates coated with poly-L-lysine and bovine serum albumin/glutaraldehyde for the binding of erythrocyte membranes and were found to be superior. We also describe the colorimetric detection of the solid phase red cell antibody reaction by using an indicator erythrocyte and peroxidase chromogenic substrate. PMID- 8249368 TI - Cold autoagglutinins with anti-Pr specificity associated with fresh varicella infection. AB - Cold agglutinins with the rare anti-Pr specificity were identified in an adult patient with fresh varicella infection. The antibody was of the IgM kappa type with subspecificity anti-Pr3d and caused a haemolytic episode in the patient. In the only previously reported case in which cold agglutinins were associated with varicella infection, the antibody specificity was also anti-Pr. PMID- 8249369 TI - Routine blood donor screening for syphilis can reveal recent infection. PMID- 8249370 TI - Renal toxicity of intravenous immunoglobulin. PMID- 8249371 TI - [Effect of penicillamine treatment on the occurrence of heart rhythm disturbances in patients with rheumatoid arthritis]. AB - An ECG examination, prolonged over several hours was carried out in 30 patients aged from 29 to 66 years (mean age 51.3) with the diagnosis of rheumatoid arthritis made on the basis of the WHO criteria, before, and in the second and third month of treatment with penicillamine. The values were evaluated of the mean, minimal, and maximal heart rate per minute, and supraventricular and ventricular heart rhythm disturbances, and also conduction disturbances were assessed. In the study no statistically significant changes were recorded in the individual periods of the examination of the values of mean, minimal and maximal heart rate/min. In the initial examination, supraventricular arrhythmia occurred in 80% of patients, and ventricular arrhythmia in 33% of cases. As a result of the applied treatment a decrease was observed of incidence of arrhythmia both supraventricular and ventricular. PMID- 8249372 TI - [Results of abdominal operations in patients with liver cirrhosis]. AB - A retrospective analysis was done of 45 patients with cirrhosis who were operated on for abdominal diseases and their complications, mainly cholelithiasis. Out of seven studied risk factors, of greatest importance were serum albumin deficiency which appeared in 45% of patients and was fraught with 100% mortality, prothrombin time (in 40%-89% mortality), and jaundice (25%-82% mortality). Postoperative complications occurred in 71% of patients with cirrhosis, mainly hepatic coma (22%), sepsis (35%), haemorrhage (18%), and eventration (22%). Postoperative mortality was 64.5%, including 76% after emergency operations, and 54% after elective surgery. After operations on the bile ducts 55% patients died. So the operations should not be performed in asymptomatic cholelithiasis during cirrhosis since there is no evidence that it deteriorates the patients' condition, and the mortality is very high. The causes are discussed of poor prognosis after surgery in cirrhosis, as well as the principles of management which may contribute to reduction of the number of complications and to a drop in mortality. PMID- 8249373 TI - [Study of acid-base balance parameters and certain electrolytes in acute diarrhea in children after oral hydration with WHO solution]. AB - The efficacy was evaluated of oral hydration with WHO solution in children with acute diarrhoea. The beneficial changes were shown of selected acid-base balance parameters and electrolytes in blood serum during this treatment. PMID- 8249374 TI - [Benign esophageal tumors]. AB - The authors present seven cases of benign oesophageal tumours treated in hospital in the years 1972-1990. Among the treated patients were six men and one woman, aged from 30 to 70 years. In four patients the tumour was situated in the middle part of the thoracic segment, and in three cases in the lower part of the supradiaphragmatic segment of the oesophagus. The diagnosis od benign oesophageal tumour was made on the basis of X-ray examination, oesophagoscopy, and histopathological examination. Five patients were treated surgically. In two patients the access to the tumour was from the right pleural cavity, and in three other cases--from the abdominal cavity. The result of histopathological examination of the removed tumours was as follows: in two patients--benign polyps, in one patient--mature teratoma, in one female patient--myxofibroma, and in one case--fibromyoma. In all patients operated on complete cure was achieved. PMID- 8249375 TI - [Preliminary clinical evaluation of unasyn preparation in genitourinary tract infections]. AB - The preparation Unasyn Pfizer in tablets of 375 mg was used for 10 days in doses of 1 tablet twice daily in 31 patients of either sex with chronic genitourinary tract infection. For criterion of treatment efficacy, the lack of significant bacteriuria was accepted as well as regression of leukocyturia and clinical signs of infection. Cure was achieved in 24 patients, improvement was found in five patients, and the lack of therapeutic effect was observed in two cases. Unasyn is useful in the treatment of genitourinary tract infections, especially by Escherichia coli and Proteus species, and is convenient in dosage, non-toxic, and well tolerated by patients. PMID- 8249376 TI - [Zinc deficiency in children infected with Giardia lamblia]. AB - The group of children infested with Giardia intestinalis consisted of 52 children aged from 3 to 10 years. The infestation was diagnosed on the basis of duodenal contents examination. The control group comprised 50 children in whom no infestation with intestinal parasites was found. Mean age of children in both groups was not different. Disturbances were found in zinc metabolism in children with Giardia intestinalis infestation. Mean serum zinc concentration in those children was 58% of serum zinc concentration in children in the control group. Similarly, zinc concentration in the erythrocytes of children in the infested group was lower and was 64% of the concentration in children in the control group. Elimination of zinc with urine in infested children was lower than in the control group, and was 36% of the level in non-infested children. Mean concentrations of serum zinc carriers, total protein, albumin fraction and transferrin were not differing between both studied group of children. Also the concentration of picolinic acid--a zinc absorption factor in the gastrointestinal tract--in duodenal contents was similar in both groups. The above mentioned results show that in children infested with Giardia intestinalis a decrease occurs of zinc absorption from the gastrointestinal tract which causes zinc deficiencies in the those children. PMID- 8249377 TI - [Psychological aspects of bone marrow marrow transplantation]. AB - Bone marrow transplantation is connected with many psychological problems of the patient determined mainly by the procedure itself. In individual phases of bone marrow transplantation (qualification of patients for operation, peritransplantation and post-transplantation periods, and final phase) different disturbances and difficulties are seen. Own observations as well as the literature data indicate the need of full diagnosis and psychotherapy, and sometimes even of psychiatric intervention during that method of treatment. PMID- 8249378 TI - [Role of viruses in pathogenesis of atherosclerosis]. PMID- 8249379 TI - [Periventricular brain edema in hydrocephalus]. AB - The authors present the pathophysiology of the zone of lowered density in periventricular white substance of the brain in computer tomography of the head in patients with hydrocephalus. The importance is discussed of that pathology for establishing of indications for surgical control of the shunt. Regression or decrease of that zone after operation is an index of efficacy of the shunt. PMID- 8249380 TI - [Zinc deficiencies in children after discontinuation of breast feeding]. AB - Zinc is a component or activator of many enzymes, especially enzymes participating in biosynthesis processes of nucleic acids and proteins. Thus the role of that microelement in the development of children is very important. The paper discusses clinical pictures of both mild and acute zinc deficiencies. Since the biological availability of zinc is much greater from mother's milk than from cow milk, thus during discontinuation of breast feeding the possibility is to be considered of occurrence of zinc deficiency. PMID- 8249381 TI - [Aztreonam--new antibiotic for treating infections caused by gram-negative aerobic bacteria]. AB - Aztreonam is the first synthetic monobactam used in practical medicine. It is effective in Gram-negative aerobe infections. It inhibits the growth of most Enterobacteriaceae in concentrations below 2 mg/ml, and of Pseudomonas aeruginosa below 16 mg/ml. It shows a widespread, distribution achieving effective therapeutic concentration there, where infections are seen most frequently. The half-life is from 1.6 to 2.0 hours. It can be administered to patients every 8 and 12 hours in single parenteral doses of 0.5, 1.0, and 2.0 g. Aztreonam is a non-toxic antibiotic, a weak hapten with slight allergenicity. It has found therapeutic use together with the antibiotics directed against aerobic and anaerobic bacterial Gram-positive flora, and it is used in the therapy directed against infections with Gram-negative aerobes. It is an effective antibiotic in nosocomial infections, in oncological patients with neutropenia, and in elderly patients. PMID- 8249382 TI - [Treatment of hemorrhoids]. AB - In the years 1980-89, 156 patients were treated in the Department of Gastrointestinal Surgery, in whom the main disease were haemorrhoids. Surgical treatment was given to 101 patients (64.74%), and 55 patients were treated conservatively (35.26%). In the group of patients treated surgically, the Whitehead method was used in 58.42% of cases, the Milligan method in 35.64% of patients, and in the remaining 5.94% the operation was carried out by other methods. Among the patients operated on, cure was achieved in 97 cases (96.04%), four patients (3.96%) required further treatment due to local complications (3 after operation by the Whitehead method, 1-by the Milligan method). More postoperative complications were seen in patients treated by the Whitehead method (5.08%) than by the Milligan method (2.78%), but for operation by the Whitehead method, patients were qualified with advanced, often complicated haemorrhoids, and all patients with accompanying anal prolapse. PMID- 8249383 TI - [Selenic enzymes as factors protecting against oxygen stress]. AB - Oxygen stress is an increase of activity of free oxygen radicals in a degree exceeding the possibility of their control by the antioxidative system of the organism. Since glutathione peroxidase was indicated as a selenium-dependent enzyme of the antioxidative system, many later studies did not confirm that function, indicating rather the participation of that protein in selenium transport. Lately, two other selenoproteins were discovered, one being a hepatic selenoprotein, participating in repairing of the damage caused by free radicals, the other being a blood serum selenoprotein which is supposed to act as a carrier. The participation of these selenoproteins in the antioxidative system of the organism is not well known. PMID- 8249384 TI - [Diagnostic difficulties and threatening complications in arterial hypertension caused by combined vascular anomaly of the abdominal aorta]. AB - A case is described of hypertension in a boy aged 8 years, admitted for elucidation of its cause is described. Abdominal aortography demonstrated a combined vascular anomaly-stenosis of the abdominal aorta and renal arteries. PMID- 8249385 TI - [Hyperthyroidism of pituitary origin]. AB - Pituitary tumour secreting thyrotropic hormone is a rare cause of hyperthyroidism. A case is discussed of hyperthyroidism of pituitary origin with coexisting valvular heart disease in the form of mitral and pulmonary valve insufficiency, and related diagnostic difficulties. PMID- 8249386 TI - [Recurrent painful ophthalmoplegia (Tolosa-Hunt syndrome)]. AB - A case is presented of recurrent painful ophthalmoplegia of atypical course, with pain in the orbit, eyelid oedema, and paralysis of external eye muscles. Disease symptoms occurred intermittently in both eyeballs. The performed MRI examination showed the presence of inflammatory focus in the posterior ethmoid. PMID- 8249387 TI - [A case of infestation with Dicrocoelium dendriticum in an 11-year old child]. PMID- 8249388 TI - [Changes of IgA, IgG and IgM immunoglobulins and of 10 other plasma proteins in patients with duodenal ulcer treated with ranitidine]. AB - In 25 patients with duodenal ulcer, before treatment and after three weeks, and another 30 days of treatment with ranitidine, plasma concentrations were determined of IgA, IgG, IgM immunoglobulins, and of 10 other proteins. In comparison to the control group of 20 persons, a statistically significant decrease was found in the concentration of alpha 2-M, prealbumins, and IgG, as well as a significant increase of alpha 1-AT, C4 complement component, and a non significant increase of coeruloplasmin, haptoglobin and IgM. During the treatment a tendency was observed for an increase of IgG and IgM level, as well as a statistically significant decrease of alpha 1-AT, C4, and coeruloplasmin concentration. The treatment exerted no effect on the decreased level of alpha 2 M. It was accepted that the tendency for an increase of IgG and IgM level was due, most probably, to the immunostimulating action of ranitidine, and the changes of concentrations of other parameters induced by the treatment were related to the presence and healing of the ulceration. The observed decrease of alpha 2-M level was probably due to binding of alpha 2-M to pepsinogen whose blood level was increased during gastric ulcer. PMID- 8249389 TI - [Value of exercise test for assessment of hemodynamic efficiency of the heart]. AB - The purpose of the work was an attempt at determination of the value of exercise test for the assessment of haemodynamic heart effectiveness. The reaction of blood pressure and heart rate to exercise was accepted as the basis for analysis. The analysis was based on a material of 40 men aged 33-66 years (mean = 51 years) with a history of myocardial infarction in whom exercise tests were carried out one year after aortocoronary bypass graft implantation. The studied men were divided into two groups A abd B depending on the reaction of blood pressure and heart rate occurring during exercise. Group A consisted of persons who responded to exercise with blood pressure and heart rate in a physiological way. The patients in group B terminated the exercise test due to pathological response. The subject of the analysis were: ischaemic changes during exercise; duration of the exercise test; attained workload in Watts; heart rhythm disturbances in group A and B; segmental left ventricular contractility disturbances evaluated on the basis of echocardiography findings. Among the patients in group B a higher percent of positive tests was found as well as shorter duration of the test and lower mean workload in relation to the patients in group A. Contractility disturbances occurred significantly more frequently - 80% among the patients in group B with greater prevalence of more sever forms of akinesis and dyskinesis (p < 0.05). Hypokinesis was observed in 7 patients (35%), akinesis in 5 cases (25%), and dyskinesis in 4 cases (20%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249390 TI - [Psychologic problems in patients with diabetes mellitus and myocardial infarction]. AB - A group was studied of 20 patients with diabetes mellitus treated in the Department of Cardiology for acute myocardial infarction. The duration of diabetes mellitus in these patients was from 5 months to 6 years, and in all patients this was the first myocardial infarction. For the study of personal features of the patients with diabetes mellitus and myocardial infarction R.B. Cattell's Self-Perception Sheet, H. Gough's ACL Adjective Test, and Multi symptomatic Diagnostic Scale MMPI-WISKAD were used. The studied showed that the group of patients with diabetes mellitus and myocardial infarction was characterized by greater intensity of depression symptoms, excessive interest in own organism and its fitness with significant manifestation of hypochondriac hysterical features and high level of restlessness and anxiety. The results of our study revealed many common features (great similarity) in the personality of patients with diabetes mellitus and myocardial infarction and patients with only diabetes but less common features with non-diabetic patients with myocardial infarction. PMID- 8249391 TI - [Comparison of beta-2-agonist and beclomethasone effects on potassium levels, heart action and pulmonary ventilation parameters in patients with chronic asthmatic bronchitis]. AB - The main goal of asthma treatment has usually been to maintain normal air potency with bronchodilators. A prompt improvement in physiologic measures of expiratory flow can be observed after administration beta-2-agonists or steroids. The present study aimed to assay the influence of fenoterol or beclomethasone therapy on the pulmonary ventilation (VC, FEV1, FEV1%), the heart action (heart rate, QT interval, 24 hours ECG) and the potassium balance in examined group. 20 patients with chronic asthmatic bronchitis were divided on two group (A and B). Group A consisted 10 subjects and patients were treated with fenoterol, the second group (Group B) consisted 10 subjects too, and patients were treated with beclomethasone. After five days treatment in all subjects an increase VC, FEV1 and FEV1% were observed. In group A (inhalation of fenoterol) we observed: 1--the significant increase of heart rate, QT interval, and frequency of cardiac arrhythmias and 2--significant decrease plasma potassium levels. In group B we didn't observed a significant changes in heart function and potassium levels. We concluded that nebulization of fenoterol, even in low doses, required control of the plasma potassium concentration and the evaluation of the heart action. PMID- 8249392 TI - [Liver microsomal enzyme function in patients with duodenal ulcer treated with cimetidine]. AB - The functional status of hepatocytes was evaluated using the breathing test with 14C-aminopyrine in a group of patients with duodenal ulcer treated with cimetidine, and the reversibility of liver microsomal system activity changes was assessed at various time intervals after the completion of treatment with the above mentioned drug. Thirty patients with duodenal ulcer treated with cimetidine were examined: before the treatment (Ia), after one day of treatment (Ib), after four weeks of treatment (Ic), after eight weeks of treatment (Id), one week after completion of treatment (Ie), and four weeks after completion of treatment. A significant decrease was found of the value of the breathing test already after one day of treatment with cimetidine which was more pronounced after four and eight weeks of treatment. An impaired microsomal system activity was shown even one week after completion of the treatment with cimetidine. The obtained results indicate the necessity of caution when other drugs are administered to patients treated with cimetidine, and this refers particularly to drugs impairing liver function, or metabolized in this organ. PMID- 8249393 TI - [Clinical evaluation of one-year controlled studies of duodenal ulcer recurrence rate in patients treated with the preparation Ulfamid]. AB - The purpose of the work was an evaluation of duodenal ulcer recurrence rate in patients with a history of at least two years of ulcer treated during one year with the preparation Ulfamid (famotidine) Krka in a maintenance dose of 20 mg before sleep. The study involved 30 patients of either sex in whom complete healing of the ulcer niche was obtained in the first phase. Control endoscopy was carried out after 3, 6, 9, and 12 months. In the first phase of the study, complete healing of the ulcer niche after the treatment with Ulfamid in a therapeutic dose of 40 mg for six weeks was obtained in 94.9%. In the second phase of the study, after three months on Ulfamid in a maintenance dose, the recurrence rate was 13.3%, after six months 26.6%, after nine months another 6.1%. After 12 months the recurrence rate of duodenal ulcer reached jointly 46.7%. Of interest is the observation of a large number of the so called asymptomatic ulcers which accounted for 50% of the observed recurrencies. PMID- 8249394 TI - [Socio-familial situation of women treated for childlessness]. AB - The documentation of the Andrology Outpatient Clinic, Institute of Obstetrics and Gynaecology, Medical Academy in Lublin from the years 1979-1989 was studied retrospectively, and 169 women were isolated who completed the treatment for childlessness. The carried out studies indicated that younger people with shorter marriage history (usually after 2-5 years) more frequently sought treatment for childlessness, especially those coming from rural areas. Greater striving for having children in married people from rural areas was expressed also by the fact that they more frequently decided to use donor semen for insemination. PMID- 8249395 TI - [Hepatic reactions in the course of viral meningoencephalitis in patients hospitalized in 1985-1989]. AB - In the years 1985-1989 in a department for parasitic and occupational diseases in Bialystok, 112 patients were hospitalized with the diagnosis of lymphocytic meningitis. The most common aetiological factor were viruses (30.6%), arboviruses (25.9%), and then herpes viruses (6.25%), and mumps virus (5.3%). In 36 patients (32%) no aetiological factor of the inflammatory process was established. In 22 patients (19.6%) a simultaneous hepatocellular damage was found in the form of jaundice (15 patients, that is 13.4%), and hepatomegaly in seven patients. The observed hepatic involvement was found in the group of patients with severe clinical course of meningitis. Most frequently hepatic reaction was observed in the group of lymphocytic meningitis caused by arboviruses. Among the biochemical indices of hepatocellular damage a significant increase in relation to normal value was found of serum bilirubin level (mean = 1.75 mg%; normally mean = 1.1 mg%), alanine aminotransferase activity was 2.5 times higher than the mean normal value (mean = 77.6 IU; normal value mean = 30 IU). The observed hepatic changes were transient and regressed together with the regression of acute inflammatory state in the central nervous system. PMID- 8249396 TI - [Comparison of doppler examination results, arteriography and intraoperative findings in patients with ischemia of the lower extremities]. AB - The diagnostic value was assessed of the examination of artery stenosis in lower extremities by the Doppler method, and the results were compared with diagnosis made by means of arteriography, and intraoperatively. The analysis included 40 patients with atherosclerosis, diabetes mellitus, and Buerger disease. Ultrasonic normal values were determined for healthy arteries, and for insignificant and significant stenosis. Treating the intraoperative diagnosis as verifying the two other methods it was found that the per cent of correct diagnosis was similar for ultrasonography and intraoperative examination, and was 76% and 66% respectively. The Doppler examination supplemented arteriography in the evaluation of crural arteries, while arteriography supplemented Doppler examination in the evaluation of the deep femoral artery and the condition of collateral blood flow. PMID- 8249397 TI - [Analysis of endometriosis cases from 10-years of surgical material]. AB - Analysis of ten years of surgical material of the histopathological research of 1865 patients population proved that in 231 cases (12.4%) patients suffered endometriosis. The ailment occurred most often in the age group 41-50 years. In general endometriosis afflicted corpus uterus (50.8% cases). In two patients the pathology was observed in abdomen wall scar after cesarean section (0.8%). PMID- 8249398 TI - [Increase of hemoglobin concentration in blood during the second trimester of pregnancy in relation to later pregnancy complications such as hypertension and intrauterine fetal growth inhibition]. AB - The concentration of haemoglobin was determined during the second trimester of pregnancy in a group of 375 women. The mean haemoglobin concentration was significantly higher in women in whom during the third trimester of pregnancy. pregnancy-induced hypertension developed (p < 0.01). Similarly, the mean haemoglobin concentration during the second trimester of pregnancy was higher in women in whom intrauterine fetal growth inhibition was found during the third trimester. The highest haemoglobin concentration during the second trimester of pregnancy was observed in women in whom pregnancy-induced hypertension developed during the third trimester, together with intrauterine fetal growth inhibition. PMID- 8249399 TI - [Comparison of endoscopic and microscopic diagnosis in patients with dyspeptic symptoms]. AB - The endoscopic and microscopic diagnosis was compared in 1030 patients with dyspeptic symptoms. The endoscopic diagnoses of "normal mucosa" and "chronic superficial inflammatory condition" were confirmed by microscopic examination in 40% of cases, and chronic atrophic inflammatory condition in 84% of cases. Gastric tumour was diagnosed endoscopically in 43 and microscopically in 47 cases. PMID- 8249400 TI - [Study of obligate anaerobic bacterial sensitivity to tinidazole and metronidazole (determination of minimal inhibiting concentration--MIC)]. AB - Comparative susceptibility testing of 428 strains of obligate anaerobic bacteria belonging to genera Propionibacterium, Arachnia, Actinomyces, Bacteroides, Prevotella, Porphyromonas, Anaerorhabdus, Fibrobacter, Fusobacterium, Peptostreptococcus and Clostridium to metronidazole and tinidazole was performed. The study of the susceptibility of anaerobic bacteria was carried out by the method of serial dilution in Brucella agar according to Finegold and Sutter (1972). Strains of B. fragilis species, B. fragilis group, other Bacteroides spp.. Fusobacterium spp. and Clostridium spp. were susceptible to both chemotherapeutics in clinically attainable concentrations. Of particular significance is the greater susceptibility of these bacteria to tinidazole. Taking into account this activity, tinidazole is a superior agent in the treatment of anaerobic infections. Both chemotherapeutics were not active against, rarely isolated from severe infections. Gram-positive anaerobic rods from genera Propionibacterium. Actinomyces and Arachnia and were partially active against peptostreptococci. PMID- 8249401 TI - [Seminoma in extragonadal locations]. AB - The pathogenesis, clinical picture and treatment of extragonadal seminoma are reviewed. For illustration a case is reported of a patient treated in the Regional Oncology Centre in Bydgoszcz. PMID- 8249402 TI - [Modern methods of treating ectopic pregnancy using surgical laparoscopic methods]. AB - The authors present modern methods of radical treatment of tubal pregnancy by the endoscopic technique. The methods are presented of conservative and radical treatment of tubal pregnancy by laparoscopy. PMID- 8249403 TI - [Pneumocystis pneumonia in infants]. AB - The acute and chronic forms are described of pneumocystis interstitial pneumonia in two infants which were observed during a hospital epidemic. The microscopic examination of bronchial secretion from the subglottic region made possible the use of guided treatment. PMID- 8249404 TI - [Leukemic infiltrations as a cause of acute renal failure]. PMID- 8249405 TI - [A case of multiple congenital anomalies of the central nervous system with neural infection and respiratory disturbances]. AB - The authors describe a case of an infant with hydrocephalus in whom respiratory disturbances occurred which were directly life-threatening. They present the differential diagnosis of this condition and put forward suggestions on aetiology and pathogenesis, based on the latest literature data. PMID- 8249406 TI - [Diagnostic difficulties in genitourinary tract tuberculosis in a 28-year old woman]. AB - The diagnostic difficulties are presented in a case of genitourinary tract tuberculosis in a 28-year-old woman. Sarcoid granulation tissue without the features of caseous necrosis which was found during histopathological examination of the tissues taken from the patient during laparotomy, suggested genitourinary tract sarcoidosis. The whole clinical picture together with the history data and laboratory investigations, and, in the first place, positive culture of the secretion from enterocutaneous fistula for Mycobacterium tuberculosis made possible the diagnosis of genitourinary tract tuberculosis. Antituberculous treatment was instituted, obtaining a significant improvement of the general and local condition. PMID- 8249407 TI - [Successful pregnancy outcome in a patient with acute renal failure superimposed on chronic renal failure treated with repeated hemodialysis]. AB - A case is presented of successful pregnancy in a patient with acute renal failure superimposed on chronic controlled renal failure. The cause which led to development of acute renal failure were severe water-electrolyte equilibrium disturbances due to diarrhoea. Appropriately conducted conservative treatment in the period of oliguria, and early institution of treatment with repeated haemodialysis (taking into account the minimal fluctuations of body fluids, and adequate heparinization) were decisive for successful outcome of pregnancy in this patient. PMID- 8249408 TI - [Effect of treatment with propranolol or pindolol on the left ventricular myocardium mass in patients with slight essential arterial hypertension]. AB - The effect was studied of 8 weeks of treatment with propranolol and pindolol on the left-ventricular myocardium mass, left-ventricular myocardium mass index, plasma renin activity, and serum aldosterone and total catecholamine concentrations. The studies were carried out in 72 patients with essential arterial hypertension of slight degree. A statistically significant decrease was shown of the left-ventricular myocardium mass, left-ventricular myocardium mass index, plasma renin activity, and aldosterone concentration in all studied patients. A decrease of total catecholamine concentration appeared in patients treated with propranolol. Weaker effect of pindolol on the studied parameters may be connected with its intrinsic sympathicomimetic activity. A correlation was shown between the decrease of the left-ventricular myocardium mass, and the decrease of systolic arterial blood pressure. PMID- 8249409 TI - [Effect of treatment with indobufen on blood viscosity indices and exercise tolerance in patients with coronary disease]. AB - The study included 20 patients aged from 50 to 65 years with stable coronary disease. In all patients exercise ECG test in Quinton moving track was performed before the treatment, and after one, three, and eight weeks of therapy. Besides that, before the treatment, and after each exercise, platelet aggregation, viscosity of whole blood and plasma, and filtration time of 1 ml of whole blood were evaluated. It was found that already after one week of treatment indobufen caused a decrease of platelet aggregation, reduction of viscosity of whole blood and plasma, and improvement of erythrocyte compliance which favourable effects increased with the continuation of the therapy up to eight weeks. At the same time as a result of eight weeks of treatment a significant improvement was achieved of exercise tolerance in the treated patients. PMID- 8249410 TI - [Certain complications of diabetes mellitus in old age]. AB - The incidence rate of the diabetes mellitus in the aged patients has been analyzed. The clinical picture, complications, and the treatment varieties were evaluated in 2260 patients observed in the diabetes dispensary. Among them this disease has been diagnosed in 500 patients after 60 year of life. The results of the observations were compared with that found in 160 patients with diabetes diagnosed before the 35 year of life. This analysis revealed that the insulin treatment decreases in the aged patients the retinopathy rate, that in them the hypersensitivity and the resistance against this hormone are rather exceptional. PMID- 8249411 TI - [Pneumonia in children and adolescents up to 18 years of age--a comparative analysis of the years 1976-1985 and 1986-1990]. AB - An analysis was done of 1084 autopsies of children and adolescents up to 18 years of age carried out in the years 1986-1990. In the analysed period 631 cases were found of pneumonia (P) (58.2% autopsies) which was an increase of about 14% in relation to the years 1976-1985. Pneumonia was most frequently observed in the age range 1-12 months. It was the direct cause of death (primary pneumonia--PP) in 130 cases (11.9% autopsies) while in 501 cases (46.2%) it accompanied other primary diseases (secondary pneumonia--SP). In relation to the years 1976-1985 a decrease was observed of the per cent of autopsies with the diagnosis of PP (from 19.5% to 11.9%), as well as a significant increase of the per cent of SP) from 25.1% to 46.2%). PMID- 8249412 TI - [Attempt at evaluation of certain parameters of gastric secretion in patients with duodenogastric reflux]. AB - There were four groups of patients: duodenal ulcer patients (32), duodeno-gastric reflux patients (31), patients with duodenal ulcer and reflux (33) and 12 healthy men. All of them aged 20-40. In all patients we performed gastroscopy, cholecystography, Kay's test for secretion and isotope proteolytic secretion test. High activity of proteolytic secretion in both groups of reflux patients was found. There were also the high percentage of false positive scores in endoscopy estimation of reflux, in comparison to the non-invasive isotope method. We suggest: reflux may be the important factor in duodenal ulcer pathogenesis; endoscopy cannot be used in duodeno-gastric reflux estimation. PMID- 8249413 TI - [Incidence of Helicobacter pylori infections in patients with duodenogastric reflux]. AB - The purpose of the work was a comparison of the incidence of Helicobacter pylori (HP) infections in persons with dyspeptic symptoms of non-ulcer dyspepsia (NUD) character in relation to the presence or absence of duodenogastric reflux (DGR). The study included 80 patients aged from 17 to 80 years, 21 males and 59 females. In all studied patients endoscopy of upper digestive tract segment was performed with biopsy specimen taking for microbiological investigations. From each patient two biopsy specimens were taken from the prepyloric part and the fundus. The specimens were subjected to a microbiological analysis using the rapid urease test, culturing, and direct preparation. The presence of duodenogastric reflux was evaluated during endoscopy. In persons with diagnosed NUD and duodenogastric reflux, lower incidence of HP infections was found than in patients with the same diagnosis but without the reflux. PMID- 8249414 TI - [Rotaviruses in acute diarrhea syndrome of infants and small children]. AB - During a period of one year 227 children aged form three days to two years were hospitalized for acute diarrhoea syndrome. In 111 cases (48.9%) the presence of HRV was observed in the faeces (group I). In 116 children (51.1%) no presence was shown of HRV in the faeces (group II). An analysis was done of the significance of differences between the observed clinical sign and abnormalities in laboratory tests in both analyzed groups. It was shown that the general condition of the children in group I was statistically more frequently moderately severe and severe (58.6% and 23.4%) than in group II. Major dehydration over 5% was found in 53.5% of cases in group I, and in 23.4% it was equal to 10% body weight. Body temperature ranged about 38 degrees C in 49.6% of children in group I while in group II in 56.9% it did not exceed 38 degrees C. Vomiting was observed very often (78.4% in group I in comparison to 35.3% in group II), especially preceding the appearance of watery faeces with mucus. The HRV diarrhoea was accompanied significantly more frequently by pharyngitis and rhinitis. Among the results of laboratory investigations of interest was greater tendency of children in group I for hypokalemia and metabolic acidosis and the white blood cell count of below 10,000, observed in 77.5% of patients. PMID- 8249415 TI - [Functional disturbances of the lower urinary tract]. AB - Six patients are described, including five women with a history of many years of refractory dysuria symptoms caused by neurosis. Four patients were elementary school teachers. In none patient changes were found in numerous general and bacteriological urine analyses nor in urography and cystoscopy, and in women also in gynaecological examination. The removal of neurosis producing factors caused a regression of dysuria symptoms in three patients. PMID- 8249416 TI - [Effect of hemodialysis on levels of parathormone in blood of patients with chronic kidney failure]. AB - In 28 patients with chronic renal failure the concentration was determined of "MID"-"carboxyterminal" PTH fragment (M/C-PTH) and of beta-2-microglobulin in the serum during haemodialysis. It was found that the serum M/C-PTH concentration was increased significantly during haemodialysis using Cuprophane dialysers. The serum concentrations of M/C-PTH and beta-2-microglobulin were not changing significantly during the first hour of haemodialysis. The authors believe that the increase of serum M/C-PTH concentration during haemodialysis may be due to unsatisfactory biocompatibility of Cuprophane dialyzing membranes. PMID- 8249417 TI - [Effect of many years of hemodialysis therapy on FSH secretion induced by luliberin in men with chronic renal failure]. AB - The purpose of the work was an explanation, what was the influence of many years of hemodialysis therapy on FSH secretion in men with chronic renal failure (CRF). The studies were carried out in 41 men with CRF treated with haemodialysis, 10 men with advanced CRF who were not dialysed, and in 15 healthy men. In all studied individuals the luliberin stimulation test (LH-RH) was performed. It was found that men with CRF, independently of the duration of hemodialysis therapy, were characterized by increased serum FSH concentration in comparison to healthy subjects. In men dialysed form over 50 months, the FSH concentration was higher than in those dialysed for a shorter time period, and not dialysed. The reaction of FSH secretion stimulated by LH-RH in men dialysed for less than 100 months and not dialysed was diminished while in those dialysed for over 100 months it was normalized. PMID- 8249418 TI - [Cholesterol crystal embolism]. PMID- 8249419 TI - [Lipid disturbances in diabetes mellitus--pathomechanism and treatment]. AB - Lipid disturbances in uncomplicated diabetes mellitus may be due to hypoinsulinaemia, hyperinsulinaemia, and hyperglycaemia. The first change is hypertriglyceridaemia with possible superimposition of hypercholesterolaemia. In diabetic nephropathy the sequence of changes is opposite. The evaluation of lipid risk factors for ischaemic heart disease in diabetes mellitus is possible only after its full controlling. The normalization of glycaemia or insulin therapy are the best methods of treatment of lipid disturbances in diabetes mellitus. PMID- 8249420 TI - [Transposition of great vessels in Cantrell syndrome]. AB - A case is presented of complete transposition of great vessels with atrial and ventricular septum defect and coarctation of the pulmonary artery in Cantrell syndrome. The Cantrell syndrome consists of: congenital heart disease, defect of pericardium, diaphragm, sternum, and anterior abdomen wall. In all cases of Cantrell syndrome described as yet ventricular septum defect was present alone or in combination with other intracardiac defects. The presented case is the first report of congenital abnormality in the from of d-TGA in Cantrell syndrome. PMID- 8249421 TI - [Isotope methods in diagnosis of portal vein occlusion]. AB - The dynamic intravenous hepato-scintigraphy with pertechnetate Tc-99m enables a quantitative evaluation of portal and arterial blood flow in the liver. A case is presented of thrombotic occlusion of the portal vein with rapidly growing oesophageal varices in which hepato-scintigraphy showed the absence of portal blood flow in the liver, being the decisive diagnostic method. Using the dynamic transrectal porto-scintigraphy an evaluation was done of the degree of collateral circulation development in the area of the inferior mesenteric vein, obtaining indirect information about localization and duration of portal vein occlusion. PMID- 8249422 TI - [Diagnostic and therapeutic difficulties in staphylococcal sepsis]. AB - A case is described of a 38-year-old man treated for three years for diabetes mellitus admitted to an internal department for elucidation of the cause of protracted fever. Staphylococcal sepsis was diagnosed with pneumonia and retroperitoneal abscess. Surgical treatment was given and targeted antibiotic therapy was administered. The patient was discharged from the hospital after 38 days as cured. An observation of 18 months failed to show the recurrence of the infection. PMID- 8249423 TI - [Hypernatremia in a patient with extrarenal kidney failure during the course of prostatic adenoma]. AB - A case is described of a rare cause of hypernatraemia in a 68-year-old patient in whom renal failure developed due to an obstacle in urine flow in the course of prostatic adenoma. Of interest are high, life-threatening values of sodium concentration in the plasma, severe course of the disease, and high grade urinary sodium loss. PMID- 8249424 TI - [Two cases of Shy-Drager syndrome--clinical picture and electrophysiological findings]. AB - Two cases are presented of Shy-Drager syndrome of various clinical symptomatology. The authors call attention to differential diagnosis and the usefulness of electrophysiological examinations in this syndrome. The particular diagnostic value of evoked auditory brainstem potential examination is stressed. PMID- 8249425 TI - [Raynaud's syndrome during the course of polymyalgia rheumatica]. AB - A case is described of polymyalgia rheumatica in which Raynaud's sign appeared at the beginning of the disease parallel with changes of rheumatoid arthritis type without other features of arterial system involvement in particular without clinical features of the involvement of the subclavian or axillary artery. The sign of temporal artery involvement appeared only after 18 months of disease duration. Steroid-therapy produced complete regression of both articular muscular and vascular changes. PMID- 8249426 TI - Comprehensive delineation of antigenic and immunogenic properties of peptides derived from the nef HIV-1 regulatory protein. AB - The Human Immunodeficiency Virus (HIV-1) nef regulatory protein, a protein involved in AIDS pathology, was used as a model to investigate and analyze B- and T-cell epitopes. In this paper, we describe the potential structural basis of antigenic and immunogenic reactivity of synthetic peptides derived from the macromolecular antigen. The relationship between B- and T-cell determinants in the context of regulatory mechanisms involved in immune recognition, while integrating recent data concerning MHC presentation. As a result of the recent progress in the field of peptide recognition and presentation, the potential of the peptide approach for constructing successful synthetic vaccines needs to be continuously re-evaluated. PMID- 8249427 TI - Testosterone impairs efficacy of protective vaccination against P. chabaudi malaria. AB - Vaccination with surface membranes isolated from Plasmodium chabaudi-infected erythrocytes can protect B10.A mice from the lethal outcome of P. chabaudi malaria. However, the efficacy depends on gender and testosterone levels. Thus, vaccination protects over 90% of female mice, but only about 55% of male mice and only about 34% of female mice when pretreated with testosterone for 4 weeks. The suppressive testosterone effect remains imprinted in females even at 10 weeks after the testosterone treatment. These data indicate that not only genetic but also environmental factors restrict the host's immune response to a malaria vaccine. PMID- 8249428 TI - Heat-shock promoter-driven synthesis of secreted bovine herpesvirus glycoproteins in transfected cells. AB - The bovine hsp70A heat-shock gene promoter was isolated and used to direct the heat-regulated synthesis of bovine herpesvirus glycoproteins gIII and gIV in transfected cultured bovine cells. Sequences encoding the viral glycoproteins incorporated mutations that deleted the transmembrane anchors. Both proteins were efficiently secreted from transfected cells in a temperature-dependent manner and the gIV so produced was found to be antigenically similar to the authentic molecule. Stable cell lines with regulated expression of these proteins were obtained and repeated thermal cycling of the cultures enabled high-yield production of these subunit vaccine antigens. The continuous production demonstrated by this system is highly relevant to the efficient and economic manufacture of vaccines and other protein biopharmaceuticals. PMID- 8249429 TI - Serological evaluation of a simplified immunization schedule using quadruple DPT polio vaccine in Burkina Faso. AB - The immunogenicity of quadruple DPT-polio vaccine used in a two-dose regimen was investigated in a cross-sectional serological survey involving 355 children under 5 years of age. This schedule is currently applied in the EPI programme in three provinces in Burkina Faso, West Africa. It was found that two doses of quadruple DPT-polio vaccine induced antibodies at protective levels to diphtheria and tetanus toxin, and to the three types of polioviruses in over 90% of 179 children. The persistence of antibodies to tetanus and polioviruses is good, since over 90% of the children studied still had antibodies more than 2 years after their last vaccination. The antibodies to diphtheria toxin tend to decline in the first 6 months after vaccination, which is not uncommon. However, our data indicate clearly that a very high percentage (98%) of children have been primed to diphtheria toxin. Of 176 non-vaccinated children, up to 25% of the older ones had antibodies to polioviruses, most of them only to one type. This appears to be a sensitive parameter for the circulation of wild polioviruses in the environment. As the vaccination coverage in the study area was low (< 60%), it was to be expected that the circulation of polioviruses in the community could not be interrupted. The present study demonstrates the applicability of a two dose strategy for primary immunization with a quadruple DPT-polio vaccine especially for poliovirus components and the toxoids. To induce pertussis immunity, however, a third vaccination is recommended. PMID- 8249430 TI - Reduction of disulfide bonds in Plasmodium falciparum gp195 abolishes the production of growth-inhibitory antibodies. AB - The role of disulfide-dependent protein conformation of the 195,000 kDa Plasmodium falciparum merozoite surface glycoprotein in the induction of biologically active antibodies was investigated. Serum samples from rabbits immunized with native gp195 had a mean ELISA titre of 1/560,000 and a mean in vitro parasite growth inhibition of 80%. In contrast, serum samples from rabbits immunized with reduced and alkylated gp195 had a mean antibody titre of 1/23,100 and did not inhibit parasite growth. These results indicate that the native structure of gp195 is essential for antigenicity, immunogenicity and induction of growth-inhibitory antibodies. Therefore, effective recombinant gp195-based vaccines may require the production of properly folded molecules resembling the native conformation. PMID- 8249431 TI - Study on the stability of 17D-204 yellow fever vaccine before and after stabilization. AB - To monitor the parameters controlling yellow fever vaccine production, eight different lots produced without stabilizers were studied. It was found that the freeze-thaw cycle did not have any adverse effect on virus infectivity and the mean loss in virus titre during lyophilization was 0.51 log10 ml-1. A thermodegradation study after storage of vaccine at different temperatures showed that the vaccine did not pass the accelerated stability test. To stabilize the 17D-204 substrain vaccine, four stabilizers with different sugars and amino acids were examined. The optimum time for addition of the stabilizers was found to be during homogenization of infected embryos. An accelerated stability test at 37 degrees C indicated that stabilizers S1, S2 and S3 kept the vaccine stable for up to 4 weeks, whereas the vaccine was only stable for up to 3 weeks in stabilizer S4. Stabilizer S3 was found statistically to be the best. Reproducibility in production methodology was established by preparing and testing more batches of the vaccine using stabilizer S3. PMID- 8249432 TI - Further studies on the adjuvanticity of stearyl tyrosine and ester analogues. AB - A new family of immunoadjuvants, long-chain stearyl esters of amino acids and peptides, are described and examined with bacterial and viral vaccines. The parent compound, stearyl tyrosine, displayed significant adjuvant activity with these vaccines. Stearyl glycyl glycine displayed superior activity with viral vaccines. A number of analogues of stearyl tyrosine were adjuvant-active. Further, these adjuvants were able to elicit a neutralizing antibody response. Stearyl tyrosine and stearyl ester analogues represent promising adjuvants for human vaccines. PMID- 8249433 TI - Mucosal immunization with a subunit respiratory syncytial virus vaccine in mice. AB - The protective efficacy of mucosal immunization with purified respiratory syncytial virus (RSV) fusion protein (F) using cholera toxin (CT) as an adjuvant was assessed in mice. Intranasal vaccination with F+CT induced F-specific serum IgG and nasal IgA and resulted in 75% protection from nasal virus replication compared with control mice. Intranasal CT+F with simultaneous parenteral F provided complete protection equivalent to intranasal live virus immunization. PMID- 8249434 TI - Monophosphoryl lipid A enhances specific CTL induction by a soluble protein antigen entrapped in liposomes. AB - Exogenous soluble antigen loaded in membranous vehicles including splenocytes and liposomes can induce a CD8+ cytotoxic T-lymphocyte (CTL) response in mice. Plain liposomes of simple composition, however, are not as effective as cellular vehicles such as splenocytes. In the present study it is shown that incorporation of monophosphoryl lipid A (MPL), a semisynthetic bacterial adjuvant, into liposomes enhanced the ability of liposomal ovalbumin (OVA) to prime for a specific CTL response. With the MPL formulation, the minimal antigen dose required for a detectable CTL induction was reduced about fivefold, and this approximated the required minimal dose of OVA loaded in the splenocytes. Moreover, liposomes containing MPL could induce a considerable level of CTL activity by either an intravenous, intramuscular or subcutaneous immunization protocol, whereas liposomes without MPL could only elicit such a response by an intravenous injection route. Subcutaneous injection of a mixture of liposomes containing MPL and liposomes containing antigen also elicited specific CTL activity. However, simultaneous subcutaneous administration of liposomal MPL and liposomal OVA at two distant sites did not prime the mice for a CTL response. These results indicate that MPL, although not necessarily incorporated in the same liposomes, must be in close proximity to the antigen to exert its adjuvant activity. Based on the results of this model antigen study, it is suggested that an optimal CTL inductive vaccine should include immunomodulatory adjuvant in addition to a class I pathway delivery vehicle such as liposomes. PMID- 8249435 TI - Diagnosis of zoster and evaluation of varicella vaccine with a passive haemagglutination assay. AB - A passive haemagglutination (PHA) assay for the detection of varicella-zoster virus (VZV) antibody was prepared with purified viral glycoproteins. Serum samples from vaccinees with live attenuated varicella vaccine, and of zoster patients, were measured for antibody titres against VZV with PHA, complement fixation (CF) and immune adherence haemagglutination (IAHA) assays, and the results compared. Antibody development could be detected as early as 3 weeks after vaccination, by both PHA and IAHA tests, but not with the CF test. Significant rises in VZV antibody in zoster patients were detected by both PHA and CF tests several days after onset. No cross-reaction was observed using HSV PHA among the vaccinees and the zoster patients. The VZV PHA assay could be used as a monitor of vaccination and a tool for differential diagnosis. PMID- 8249436 TI - Vaccination with two different vaccinia recombinant viruses: long-term inhibition of secondary vaccination. AB - The effects of immunity to vaccinia virus on the efficiency of vaccination with a vaccinia recombinant virus were studied. In mice the efficiency and duration of the B-cell response to the recombinant gene product of a second vaccinia recombinant virus were suppressed for more than 9 months, i.e. practically lifelong. Antibody titres against the recombinant gene product were not only lower but also lasted for a shorter time. Thus, immunity to vaccinia virus may influence both the titre and duration of the antibody response induced by a second distinct vaccinia recombinant vaccine. PMID- 8249437 TI - Use of single-strand conformation polymorphism technique for the initial characterization of virus isolates. PMID- 8249438 TI - Immunoadjuvant activities of liposomes: two different macrophage-mediated mechanisms. PMID- 8249439 TI - Poliovirus neurovirulence assays by ulnar nerve inoculation in bonnet monkeys. PMID- 8249440 TI - WHO/OIE meeting: consultation on newly emerging strains of equine influenza. 18 19 May 1992, Animal Health Trust, Newmarket, Suffolk, UK. PMID- 8249441 TI - Cell immortalization: the role of viral genes and carcinogens. PMID- 8249442 TI - Classification of a new member of the TBE flavivirus subgroup by its immunological, pathogenetic and molecular characteristics: identification of subgroup-specific pentapeptides. AB - The antigenic, pathogenic and molecular characteristics of Turkish sheep encephalitis (TSE) virus, strain TTE80, were compared with other members of the tick-borne encephalitis (TBE) virus complex. Monoclonal antibodies with defined specificity for the flavivirus envelope glycoprotein distinguished TSE virus from louping ill (LI), western or far eastern TBE, Langat and Powassan virus in indirect immunofluorescence, haemagglutination-inhibition and neutralization tests. On the other hand, TSE virus, which produces an LI-like disease in sheep, resembled LI virus in mouse neurovirulence tests. Molecular homology data of all the structural genes of TSE virus compared with other tick-borne flaviviruses demonstrated that TSE virus is a distinct member in the TBE virus subgroup. The data are consistent with the conclusion that TSE virus has evolved by a separate evolutionary pathway as compared with the close antigenic relatives, western European, far eastern TBE viruses and LI virus. By aligning the encoded amino acids in the viral envelope glycoprotein of mosquito- and tick-borne flaviviruses, we have also identified subgroup-specific pentapeptide motifs for the tick-borne encephalitis, Japanese encephalitis and dengue subgroup viruses of the genus Flavivirus. These pentapeptides have important implications for the evolution, classification and diagnosis of flaviviruses. PMID- 8249443 TI - Nucleolar accumulation of core protein in cells naturally infected with Semliki Forest virus. Quantitative aspects. AB - The core (C) protein of Semliki Forest virus (SFV) is known to exert several important functions with regard to both the virus and the host. This paper shows that migration of parental and progeny C protein to the nucleolus is a common feature in infected vertebrate and invertebrate cells. The amount of C protein accumulating to the nucleolus is small, always less than 1% of the intra- and extracellular C protein at various times post infection. Migration to the nucleolus is a fast process; 1.5 h post infection a prominent amount of parental C protein is already incorporated into nucleolar fractions. PMID- 8249444 TI - Comparison of nucleotide sequences among hantaviruses belonging to the same serotype: an analysis of amplified DNA by thermal cycle sequencing. AB - The hantavirus genus, belonging to the bunyaviridae family, is comprised of at least four serologically distinct types: Hantaan, Seoul, Puumala and Prospect Hill. Previously, we reported the use of the polymerase chain reaction (PCR) for grouping hantavirus isolates by using four sets of primers specific to each serotype. Our PCR typing results agreed with those of serological typing. The present study makes use of thermal cycle sequencing to sequence PCR-amplified DNA products in order to determine the level of similarity among members of the same serotype. We show that members of Hantaan and Seoul serotypes are over 92% homologous, irrespective of their host and geographical origin. Puumala sequences show a degree of homology ranging from 80 to 98%. Despite the variation in sequence at the nucleotide level, amino acids show an even higher level of conservation. PMID- 8249445 TI - Reconstitution of wild type viral DNA in simian cells transfected with early and late SV40 defective genomes. AB - The DNAs of polyomaviruses ordinarily exist as a single circular molecule of approximately 5000 base pairs. Variants of SV40, BKV and JCV have been described which contain two complementing defective DNA molecules. These defectives, which form a bipartite genome structure, contain either the viral early region or the late region. The defectives have the unique property of being able to tolerate variable sized reiterations of regulatory and terminus region sequences, and portions of the coding region. They can also exchange coding region sequences with other polyomaviruses. It has been suggested that the bipartite genome structure might be a stage in the evolution of polyomaviruses which can uniquely sustain genome and sequence diversity. However, it is not known if the regulatory and terminus region sequences are highly mutable. Also, it is not known if the bipartite genome structure is reversible and what the conditions might be which would favor restoration of the monomolecular genome structure. We addressed the first question by sequencing the reiterated regulatory and terminus regions of E- and L-SV40 DNAs. This revealed a large number of mutations in the regulatory regions of the defective genomes, including deletions, insertions, rearrangements and base substitutions. We also detected insertions and base substitutions in the T-antigen gene. We addressed the second question by introducing into permissive simian cells, E- and L-SV40 genomes which had been engineered to contain only a single regulatory region. Analysis of viral DNA from transfected cells demonstrated recombined genomes containing a wild type monomolecular DNA structure. However, the complete defectives, containing reiterated regulatory regions, could often compete away the wild type genomes. The recombinant monomolecular genomes were isolated, cloned and found to be infectious. All of the DNA alterations identified in one of the regulatory regions of E-SV40 DNA were present in the recombinant monomolecular genomes. These and other findings indicate that the bipartite genome state can sustain many mutations which wtSV40 cannot directly sustain. However, the mutations can later be introduced into the wild type genomes when the E- and L-SV40 DNAs recombine to generate a new monomolecular genome structure. PMID- 8249446 TI - Changes in expression of surface and core antigens of hepatitis B virus in different mutant clones of hepatoma PLC-PRF-5 cells. AB - Mutant clones of human hepatocarcinoma PLC-PRF-5 cells carrying a hepatitis B virus (HBV) genome have been obtained using selection for resistance to the toxic action of a variety of preparations to induce cell differentiation. The clones differed in various features such as expression of alpha-fetoprotein (AFP) and albumin as well as in growth rates, ability to grow in semisolid media and to be cloned in agar. Expression of the surface antigen (HBsAg) was significantly increased in mutant cells exhibiting differentiation features in contrast to the parental cells. In addition, the core antigen (HBcAg), which was silent in the original cells, was detected in some clones. There was no temporal correlation between the peak of enhanced expression of HBsAg and activation of HBcAg observed at different life periods of each clone. Evidence of cell fusion in cell culture such as premature chromatin condensation and increased numbers of binucleate cells was detected in clones with differentiation features and an increased level of viral gene expression. The approach used in this study can be used to develop cell lines of the same origin with different degrees of differentiation whilst maintaining HBV expression. This model system may be useful in the study of HBV. PMID- 8249447 TI - Cymbidium mosaic virus RNA synthesis in isolated orchid protoplasts. AB - An in vitro protoplast host system has been developed to facilitate the study of cymbidium mosaic virus (CyMV) replication mechanisms. CyMV, a member of the potexvirus group of plant viruses, is among the most significant pathogens of cultivated orchid plants, but a convenient system for studying its molecular genetics has not been available previously. A cDNA probe representing a segment of the CyMV genome was used to detect viral RNA following inoculation of Vanda floral protoplasts with virus particles. Detection was carried out by RNA-DNA hybridization methods (RNA slot blots). Newly synthesized viral RNA was detectable by 6-9 h postinfection and appeared to reach a maximum around 12 h. These results are consistent with the time course of viral RNA synthesis in protoplasts infected with other RNA viruses. Immunoblot detection methods revealed the presence of newly synthesized viral protein indicating that viral gene expression occurs in the protoplasts in addition to genome replication. PMID- 8249448 TI - Workshop on 'reverse genetics of negative stranded RNA viruses' sponsored by the Juan March Institute, Madrid, Spain. PMID- 8249449 TI - [Brain monoamine oxidase and the predisposition for catalepsy in tame and aggressive Norway rats]. AB - Aggressive Norway rats predisposed to catalepsy were characterized by an increased MAO B and dopamine-deaminating activities in the brainstem as compared to non-predisposed ones, MAO A activity being unchanged. Catalepsy-susceptible grey rats from a substrain of the tamed animals had lowered MAO A, MAO B, and dopamine-deaminating activities in the brainstem in comparison with the nonsusceptible ones. Among catalepsy-non-susceptible rats from both aggressive and tame populations, the brainstem MAO A and MAO B activities were considerably higher in tame animals than in aggressive ones. PMID- 8249450 TI - [A multiparameter method for the complex evaluation of anxiety-phobic states in rats]. AB - A new multiparameter method is elaborated for evaluating the anxiety-phobic states in rats. The method is based on a ranged scale of parameters which characterize the species-specific responses of an animal to the series of ethologically adequate test-stimuli inducing manifestation of the anxiety-phobic states. The method makes it possible to evaluate an individual anxiety-phobic level of a rat, to form experimental groups of rats with known individual anxiety phobic levels, to conduct repeated observations of the same animals. The method is easy to introduce into laboratory practice and makes it possible to obtain results quickly. PMID- 8249451 TI - [The motor organization of the sense of rhythm]. PMID- 8249452 TI - [The 3-dimensional localization of the afterdischarges of single visual evoked potentials]. PMID- 8249453 TI - [The synaptic release of dopamine in the striatum of rats during the acquisition of a passive avoidance conditioned reflex]. AB - To study the striatal synaptic dopamine release during passive avoidance behaviour the intracranial microdialysis technique combined with radioenzymatic assay method was used in Sprague-Dawley rats. It was shown that the acquisition of passive avoidance conditioning and an unescapable footshock led to delayed increases in striatal synaptic dopamine release. PMID- 8249454 TI - [The reaction of rats with active- and passive-type behaviors during the "collision" of 2 unconditioned reflexes]. PMID- 8249455 TI - [The spontaneous behavior of rats and the immobilization stress-evoked accumulation of TBA-active products in the tissues: a cluster analysis]. PMID- 8249456 TI - [Have command neurons in the CNS of the snail been found?]. PMID- 8249457 TI - [The regulations of the Svetloe Piatno International Research Center on the Neurobiology of Consciousness]. PMID- 8249458 TI - [The brain in stereotaxic coordinates (a textbook for colleges)]. AB - The present textbook is directed forward students of universities and medical colleges, young scientists and practicing doctors dealing with stereotaxic method. The Paxinos and Watson stereotaxic rat brain atlas (1982) is the basis of the textbook. The atlas has been transformed into computer educational program and seven laboratory works: insertion of the electrode into brain, microelectrophoresis, microinjection of drugs into brain, electrolytic destruction in the brain structures, local brain superfusion. The laboratory works are compiled so that they allow not only to study practical use of the stereotaxic method but to model simple problems involving stereotaxic surgery in the deep structures of brain. The textbook is intended for carrying by IBM PC/AT computers. The volume of the textbook is 1.7 Mbytes. PMID- 8249459 TI - [The negative components of visual evoked potentials in the mechanisms of the classification of incomplete images]. AB - Event-related brain potentials were recorded in 16 young adult subjects in response to a short presentation of recognizable and nonrecognizable visual images with an incomplete set of signs. Sequential brain maps and amplitude temporal ERP parameters in 1000 ms poststimulus epoch were estimated (BMDP 4M and 7M, 1987). The results suggested that the component N450 was specifically related to a perception of nonrecognizable visual images. N70 and N150 reflected voluntary attention during invariant recognition and the late negative waves (600 1000 ms) reflected an "exhaustive search" for categorization of new information. PMID- 8249460 TI - [Endogenous cortical evoked potentials in parkinsonism]. AB - Averaged cortical evoked potentials (EP) to light stimuli were studied in 19 Parkinsonian patients and 12 healthy subjects under the conditions of passive perception ("the background") and during different psycho-motor reactions. Differential EP waves between "the reaction" and "the background" were analysed, i.e. the endogenous potentials. A retardation of the psycho-motor reactions in patients was found to be accompanied by a decrease of endogenous EP in the frontal recording. This decrease mainly concerned the potentials connected rather with the motor acts than with the cognitive operations. The retardation of the voluntary movements in parkinsonism is assumed to be connected with a "deficit" of the frontal activation due to a disfunction of the basal ganglia. PMID- 8249461 TI - [Teaching monkeys complex behavioral tasks]. AB - Differences of two ways of training were shown in two monkeys. Random order of alteration of two conditioned signals was extremely ineffective. The use of a special algorithm of signals' administration dependent on the behaviour of a monkey made the learning of complicated tasks possible. Probable mechanisms of the effect described are discussed. PMID- 8249462 TI - [The participation of the projection areas of the temporal cortex in the sensory support of certain forms of cognitive processes]. AB - In neuropsychophysiological and biochemical experiments was studied the role of the temporal cortex of the cat (AI, AII) in mnemonic, perceptive, gnostic functions, praxis, and the higher cognitive processes. The participation of the temporal fields in the mechanisms forming the gnostic imagery activity was shown. PMID- 8249463 TI - [The bilateral interaction of the hypothalamus, amygdala and neocortex at different stages in conditioned reflex acquisition]. AB - By averaged evoked potentials of the symmetric points of the cat's auditory cortex (Al), hypothalamus (Hl) and amygdala (Abl) in response to a sound was determined the character of the two-way interaction (Spearman coefficient of correlation) between the structures before and at different stages of instrumental food-procuring conditioning. Individual character of co-tuning of Al, Hl, and Abl as well as the emotional state connected with adaptation of an animal were found to be reflected in the correlation of evoked activity of the structures. Conditioning led to a formation of close interconnections between the structures eliminating individual peculiarities of correlation. Stabilization of the response was accompanied by a weakening of the interconnections between the cortex and the subcortical structures, internal symmetrical co-tuning of Al, Hl and Abl being unchanged. PMID- 8249464 TI - [The dynamic characteristics of the chaotic component of the high-frequency EEG of the rabbit during the acquisition and extinction of the orientation reflex]. AB - The chaotic component of the high-frequency EEG of the rabbit was studied during repeated presentation of non-rhythmical light stimuli. The chaotic component was found to be sensitive to the action of these stimuli, its reactions being variant in different cortical areas. Dimension changes were revealed corresponding to a manifestation of the phasic and tonic components of the orienting reaction. In the course of orienting reflex extinction the chaotic component characteristics changed in correspondence with the activation level shifts in the cortical areas under study. PMID- 8249465 TI - [The effect of intraspecific isolation in early ontogeny on the formation of a local instrumental reflex in male and female rats]. AB - Peculiarities of local instrumental conditioning were studied in male and female rats brought up under the conditions of intraspecific isolation and in group. Early intraspecific isolation was found to cause a failure of the local instrumental conditioning in male rats but not in females. Right-handed and ambidextrous males brought up in isolation were shown to adapt to a new situation and to form a motor habit slower than the animals brought up in group. This occurred due to an increase of emotional reactivity of isolants. The early intraspecific isolation did not influence the velocity of the motor habit formation in males with motor dominance of the right hemisphere. In male and female rats was revealed the different sensitivity of the brain to a deficit of biologically significant influences in early ontogeny. PMID- 8249466 TI - [Prolonged homosynaptic depression of the impulse reactions of the motor cortex neurons in the cat]. AB - It is shown that in the motor and the visual cortices of the cat the homosynaptic long-term posttetanic depression (LTD) of monosynaptic impulse responses of the cortical neurons may be induced in the tetanized input. Homosynaptic LTD appears as a decrease of the probability of the monosynaptic discharges or an increase in the latency of the monosynaptic responses. The cortical homosynaptic depression possesses the same properties as the hippocampal LTD, namely, the longevity, input specificity, cooperativity, and associativity. The possible mechanisms of the homosynaptic LTD induction are discussed. The effect may be determined, on the one hand, as Ca-dependent phenomenon, and on the other hand, as the LTP of monosynaptic reactions of the input inhibitory interneurons. It is supposed that the homosynaptic LTD of the impulse reactions of the cortical neurons may be one of the basic mechanisms in certain learning tasks, such as habituation or extinction. PMID- 8249467 TI - [The effect of moderate doses of ethanol on the nature of the background neuronal impulse activity of the sensorimotor cortex in rats]. AB - The influence of acute administration of moderate doses of ethanol (170-1200 mcg/kg, intraperitoneally) on the spontaneous impulse activity of motor cortex neurons was studied in outbred rats never earlier exposed to alcohol. Initial reactions of the cortical neurons were shown to be characterized by the changes in the frequency of spontaneous activity which could be increased or decreased during the first 20-25 min after the injection. The changes in the mean frequency of spontaneous discharges were accompanied by the changes in their patterns. Both increase and decrease of the mean frequency at the initial stage of ethanol action was accompanied by significant variability of interspike intervals. It is emphasized that examination of the initial reactions of the cortical neurons to ethanol administration demands studying their temporal duration. PMID- 8249468 TI - [The role of cAMP in the electrophysiological effects of morphine and enkephalins]. AB - The review considers the data on the morphine and enkephalins electrophysiological effects which were obtained in snail neurons and might be explained by a decrease in the level of cAMP in the cytoplasm. These effects are: a membrane hyperpolarization, a depression of the bursting activity, an inhibition of the excitatory serotonin and dopamine responses. PMID- 8249469 TI - [The role of second messengers in the modulation of the plasticity of the cholinoreceptors of the snail RPa3 neuron by the opiate kappa agonist bremazocine]. AB - In the paper was shown the participation of some second messengers in intracellular molecular mechanism of a decrease of RPa3 neuron cholinoreceptor plasticity by opiate kappa-agonist bremazocine during rhythmical acetylcholine application. This mechanism includes 1) inhibition of inositol-1,4,5 trisphosphate-dependent mobilization of deposited Ca2+ activating intracellular receptor calmodulin and 2) inhibition of the synthesis of the acyclic eicosanoids. It is possible to assume a specificity of this mechanism, so far as is not revealed the role of the second messengers under study in bremazocine inhibitors of RPa3 neuron somatic cholinoreceptors under the conditions of non rhythmical acetylcholine application. PMID- 8249470 TI - [The neurochemical correlates of plasticity]. AB - Water-soluble proteins in the CNS of Helix pomatia were studied by the method of disk electrophoresis in polyacrylamide gel. The protein Rf 0.58 proved to be absent in young snails, while in adults its amount varied from 3% (naive snails) to 6-10% (trained animals) of the proteins separated by the method used. Incubation of the adults' ganglia in saline containing serotonin or NaF led to a significant elevation of the protein Rf 0.58 quantity. Neither serotonin nor NaF induced the protein Rf 0.58 in the young snails. We suggest the protein Rf 0.58 to be related to G-proteins. Late maturation of G-proteins might be one of the reasons of low behavioural plasticity in the young snails. Influence of acetylcholine on the protein synthesis in the identified "pool D" neurons was studied too. Protein fractions were identified, in which the intensity of labelling increased up to 400-500% of the control level. Acetylcholine is supposed to modulate the protein synthetizing system in these neurons. PMID- 8249471 TI - [Receptors of ionotropic mediators in snail neurons: the ionic nature and mechanisms of their functional regulation and topography]. AB - In the paper are described the responses of identified neurons to application of acetylcholine, glutamate, glycine, and gamma-aminobutyric acid. Were studied the ionic mechanisms of these responses and the effects of different agents (metabotropic neurotransmitters, peptides, vitamins, and lectins) on the functional properties and topography of the receptors of ionotropic neurotransmitters. PMID- 8249472 TI - [A method for assessing the functional state of an animal (cat) during the modelling of neurotic pathology]. AB - The study was aimed to accumulate and classify the data on different degrees of experimental neurotic pathology. Experiments were carried out with cats using the method of food-procuring conditioning. Behavioural, vegetative reactions and a set of electrophysiological characteristics were recorded. The experimental neurotization was evoked by different factors: acute emotional stress, chronic stress, artificially increased and lowered brain activation. Classification of the facts obtained allowed to create a scale of parameters corresponding to different degrees of pathology (weak, middle, deep). Every characteristic may be quantitatively estimated using the scale. The summarized evaluation was used to determine the functional state of an experimental animal. PMID- 8249473 TI - [The long-term memory and learning capacity of rats with high brain seizure activity after an embryonic nerve tissue allograft]. AB - Fragments of the embryonic hippocampus, septum, cerebellum and substantia nigra were grafted into the lesioned brain areas of 151 animals with phenotypical (bilateral destruction of the CA1 and CA4 hippocampal areas or the basolateral amygdalar regions in Wistar rats) and genotypic (in Krushinskii-Molodkina strain) high seizure readiness. The parameters of long-term memory were improved after embryonic septal and cerebellar graftings, normalized after embryonic hippocampal graftings and were not improved after embryonic nigral graftings in both strains. Learning ability of the animals of both the strains with ceased audiogenic seizures was kept at the control level after the transplantation of embryonic tissue from the different brain areas. In animals with continued seizures the learning ability normalized after the septal graftings. PMID- 8249474 TI - [The comparative characteristics of the work capacity of adult and older rats under positive and negative emotional reinforcement]. AB - Age-dependent characteristics of motor activity during self-stimulation and active avoidance of stimulation of the hypothalamus were studied in rats. Capacity for work of the adult rats was found to be higher than that of the old ones during the positive as well as the negative emotional reinforcement. Age dependent differences of the capacity for work were more expressed during the negative emotional reinforcement being connected with relatively fast destruction of motivational behaviour in the old animals. PMID- 8249475 TI - [Identification of irradiated pasteurized egg products: a combined method for use in routine control]. AB - Pasteurized egg products (whole egg, egg yolk and egg white) were tested for irradiation treatment in the German food control laboratories in Oldenburg/Niedersachsen and Kassel/Hessen as well as in the food irradiation laboratory of the German federal health office. Gas chromatographic/mass spectrometric measurements on the fat components of egg-products showed clearly whether the product had been irradiated or not. While in unirradiated samples no traces of special hydrocarbons (according to the fatty acid composition of egg) and no traces of the irradiation-specific compound 2-Dodecyl-cyclobutanone were found, irradiated control samples as well as products of two Belgian suppliers contained these substances. Additionally, regarding the rather high time consumption of gas chromatography, electron spin resonance (ESR)-measurements were carried out on the packaging material of egg products. Irradiated packaging material (cellulose) could be easily detected by the appearance of a signal pair in the ESR spectrum (cellulose radical). ESR measurements are very fast and easy to perform so that this method can be used for screening. Microbiological investigations showed remarkably reduced total numbers of microorganisms for some irradiated samples, but the microbiological status is influenced by other factors like storage-time and -temperature, so that microbiological tests can not be used successfully for screening on irradiation treatment. PMID- 8249476 TI - Contribution of aluminum from packaging materials and cooking utensils to the daily aluminum intake. AB - Migration of aluminum (Al) from packaging materials and cooking utensils into foods and beverages was determined at intervals during cooking or during storage by graphite furnace atomic absorption spectroscopy. High amounts of Al migrated into acidic products such as mashed tomatoes during normal processing in normal, non-coated Al pans. After 60 min cooking an Al content of 10-15 mg/kg was measured in tomato sauce. Surprisingly, the Al concentration was also increased up to 2.6 mg/L after boiling tap water for 15 min in Al pans. Storage of Coca Cola in internally lacquered Al cans resulted in Al levels below 0.25 mg/L. In contrast, non-coated Al camping bottles containing lime blossom tea acidified with lemon juice released up to 7 mg Al/L within 5 days. The Al concentration in coffee was lower than that of the tap water used in its preparation, even if prepared in Al heaters. In Switzerland, where most pans nowadays are made of stainless steel or teflon-coated Al, the average contribution for the use of Al utensils to the daily Al intake of 2-5 mg from the diet is estimated to be less than 0.1 mg. PMID- 8249477 TI - Evaluation of an in vitro method for the estimation of the selenium availability from cow's milk. AB - The present study evaluates the application of an in vitro digestion procedure for estimating the selenium (Se) availability from cow's milk. The results obtained by this in vitro method indicate that only a small portion (< 10%) of milk Se can be estimated as available. The available fraction of whole milk (4.7%) was significantly lower compared to the Se availability of skim milk (7.3%). Investigations on the methodology of the in vitro method indicate that the portion of total potentially available Se, determined as total dialysable Se, increases simultaneously with protein digestibility along with the digestion process. Therefore it is concluded that protein digestibility might largely influence the Se availability from milk. PMID- 8249478 TI - Effects of shooting period, times within shooting periods and processing systems on the extract, caffeine and crude fiber contents of black tea. AB - The extract, caffeine and crude fibre contents of black tea, from different shooting periods, different times within each shooting period and processed by five different commercial rolling methods, were examined. The extract and crude fibre contents of black tea were significantly (P < 0.01) affected by all these factors and their interactions. The caffeine content of black tea was affected by the shooting period and times within the shooting period. It was determined that processing methods did not have a significant (P < 0.05) effect on the caffeine content. The amount of extract and caffeine decreased from the first shooting period to the third, and also from the beginning of each shooting period to the end. However, the crude fibre content of the black tea increased, especially from the beginning of each shooting period to the end of each shooting period. PMID- 8249479 TI - [Contamination of hazel nuts and chocolate by mineral oil from jute and sisal bags]. AB - Before spinning, jute and sisal fibres are treated with a batching oil commonly consisting of a raw mineral oil fraction. Such oil is transferred to foods packed into jute or sisal bags, i.e. primarily cocoa beans, hazelnuts, coffee, almonds, oil seeds, and rice. Concentrations in the foods easily exceed 100 mg/kg. Chocolate may be affected several-fold: cocoa beans as well as hazelnuts and almonds may be contaminated. PMID- 8249480 TI - Influence of sous vide processing, steaming and boiling on vitamin retention and sensory quality in broccoli florets. AB - In this study the effect of sous vide processing, steaming and traditional boiling of broccoli florets on retention of ascorbic acid, vitamin B6 and folacin was investigated. Freshly prepared samples were also sensorily evaluated. In all heat treatments ascorbic acid was found to have the highest retention, and was therefore judged not to be suitable as an indicator for vitamin retention in general. When the three types of heat treatment were compared, boiling showed lowest retention of all the vitamins examined (45-64% in 5 min), whereas sous vide processing gave the highest retentions (97-100% in 5 min). Retentions during steaming were a little lower (83-100% in 5 min). Sensory evaluation showed that sous-vide cooked and steamed broccoli florets generally had higher acceptability than boiled. PMID- 8249481 TI - [The amino acid composition of the edible Russula and Agaricus mushroom species]. AB - The amino acid concentrations of six samples (five species) of the genus Russula and of seven samples (five species) of the genus Agaricus were determined. The Russula samples have significantly lower total amino acid concentrations (14.79% of the dry weight), than the Agaricus species (23.75%). Essential amino acids occur in both groups in appreciable concentrations: 48.9% of the total amino acid concentration for Russula and 46.4% for Agaricus. Between the cultivated varieties and the wild species of Agaricus are insignificant differences in the total amino acid concentration, but the quantities of certain essential amino acids were lower, whereas the concentration of Glu was higher in the wild species. PMID- 8249482 TI - Organochlorine residues in the edible part of eels of different origins. AB - Fifty-four eel samples from 11 different countries were analysed by capillary gas chromatography for levels of the pesticides aldrin, endrin, hexachlorobenzene, alpha- and beta-BHC, lindane, heptachlor, octachlorostyrene, pp'-DDT, pp'-DDD, pp'-DDE, and for the levels of the PCB congeners 28, 52, 101, 138, 153, and 180. Wild eels, especially from the Baltic Sea area, had elevated contents compared to farmed eels. The mean contamination level reached only 1-5% of the allowable values given by German directives and never exceeded the official limits. PMID- 8249483 TI - [Analysis of element species and the question of the bioavailability of calcium from different types of bread]. AB - In NaCl and NaCl/HCl extracts from 11 kinds of bread and flour the calcium ion concentrations in comparison to the total calcium contents were determined by a calcium-selective electrode. Crisp-breads showed the slowest values of extractable calcium and calcium ion concentrations in the extracts. Differentiated investigations on calcium binding in the neutralized extracts were carried out with a view to studying calcium resorption in the duodenum. Correlations with the phytic acid contents have shown that other degradation products also take part in the re-binding of calcium. By these investigations the contribution of element speciation to the evaluation of calcium bioavailability has been shown by the analytical methodology and the results of application. PMID- 8249484 TI - Annexins--multifunctional, calcium-dependent, phospholipid-binding proteins. PMID- 8249485 TI - Factors controlling the activity of serine- and other base-exchange enzymes and phosphatidylserine transport in mammalian cell. PMID- 8249486 TI - ATP-dependent phosphatidylserine formation in animal cells as a base exchange reaction. PMID- 8249487 TI - Thapsigargin: potent inhibitor of Ca2+ transport ATP-ases of endoplasmic and sarcoplasmic reticulum. PMID- 8249488 TI - Calcyclin--from basic research to clinical implications. PMID- 8249489 TI - ATP-sensitive K+ channel in mitochondria. PMID- 8249490 TI - Transport and metabolic effects of carnitine and acylcarnitines in brain. PMID- 8249491 TI - The significance of myosin light chains in mechanochemical coupling in skeletal muscle. PMID- 8249492 TI - Motility assay: achievements and perspectives. PMID- 8249493 TI - Specificity of thymidylate synthase inactivation by 4,5-bisubstituted dUMP analogues. PMID- 8249494 TI - Synergistic cell growth inhibition by combination of antifolates. PMID- 8249495 TI - Polyamines as regulators of cell activation. PMID- 8249496 TI - Differential role of polyamines in hyperplasia and hypertrophy. PMID- 8249497 TI - Transcription factors in cellular senescence and death. PMID- 8249498 TI - Gangliosides in the repair of brain cholinergic neurons. PMID- 8249499 TI - Subcellular localization of two different type-1 casein kinases from yeast. AB - Specific antibodies directed against the two yeast type-1 casein kinases (CK1) were used to study the localization of both 45 kDa and 27 kDa casein kinase species in yeast cells by immunofluorescence. Our results indicate that the larger and smaller CK1 species are localised in different compartments of the yeast cell. The 45 kDa enzyme is present in the cytoplasm of the cell both during the logarithmic and stationary growth phase. The 27 kDa CK1 was found in the nucleus in the cells in logarithmic growth phase while the enzyme from the stationary phase was present in the cytoplasm. Our results suggest that the 27 kDa casein kinase may play some role in yeast cell division control by displacement from the nucleus to the cytoplasm. PMID- 8249500 TI - Comparison of the kinetic properties of the pyruvate dehydrogenase complex from pig kidney cortex and medulla. AB - The activity of the pyruvate dehydrogenase complex (PDC) purified from pig kidney medulla was affected by K+, Na+, Cl-, HCO3-, HPO4(2-) and changes in ionic strength. Increased ionic strength influenced the activity of PDC from medulla by decreasing the Vmax and S0.5 for pyruvate and increasing the Hill coefficient. The magnitude of these changes was smaller than the corresponding changes for PDC purified from the cortex. In the presence of K+ (80 mM), Na+ (20 mM), Cl- (20 mM), HCO3- (20 mM), HPO4(2-) (10 mM) and at ionic strength of 0.15 M the S0.5 for pyruvate of PDC from medulla was 117 microM and the enzyme complex was saturated by 1.1 mM pyruvate. Under these conditions the S0.5 for pyruvate of PDC derived from cortex was 159 microM and the enzyme was saturated at 4.5 mM pyruvate. Based on the results presented in this report it is suggested that PDC in kidney medulla may be regulated not only by a phosphorylation/dephosphorylation system and end-product inhibition but also via changes in ionic strength. PMID- 8249501 TI - Comparative studies on O-acetylhomoserine sulfhydrylase: physiological role and characterization of the Aspergillus nidulans enzyme. AB - O-acetylhomoserine sulfhydrylase (OAH SHLase) from Aspergillus nidulans is an oligomeric protein with a broad substrate specificity with regard to sulfhydryl compounds. As its Saccharomyces cerevisiae counterpart the enzyme also reacts with O-acetylserine and is inhibited by carbonyl reagents but not by antiserum raised against the yeast enzyme. In contrast to Saccharomyces cerevisiae the enzyme is not essential for Aspergillus nidulans as indicated by the completely prototrophic phenotype of OAH SHLase-negative mutants. Its major physiological role in Aspergillus nidulans seems to be recycling of the thiomethyl group of methylthio-adenosine but it is also a constituent of the alternative pathway of cysteine synthesis. PMID- 8249502 TI - Diagnostic assessment of enlarged superficial lymph nodes by fine needle aspiration. AB - Two hundred eighty-five consecutive outpatients with enlarged superficial lymph node either clinically suspicious (152) or with a previous diagnosis of a malignant tumor (133) underwent fine needle aspiration (FNA) followed by excisional biopsy. Cytologic and/or cytologic-immunophenotypic diagnoses made on direct smears were compared with subsequent histologic findings. The comparison demonstrated (1) a high rate of conclusive cytologic diagnoses in the assessment of metastatic malignancies, with an overall accuracy rate of 99.1% and a typing accuracy rate of 96.5%; (2) a high rate of conclusive diagnoses in the assessment of high grade non-Hodgkin's lymphomas and Hodgkin's disease, with the exception of the lymphocytic predominance variant of the latter; and (3) significant limitations in the assessment of low grade non-Hodgkin's lymphomas because of the high rate of false-negative diagnoses in cases with a substantial nonmalignant cell component. This was particularly evident in follicular centroblastic centrocytic lymphomas. Immunocytochemistry appeared to be of limited value in the distinction between centroblastic-centrocytic follicular lymphomas and reactive follicular hyperplasia. The results confirmed the diagnostic value of fine needle aspiration as the first step in the workup of patients with nodal enlargement suspicious for malignancy. In the area of low grade non-Hodgkin's lymphomas, morphologic and immunocytochemical methods need to be supplemented by molecular techniques in order to achieve conclusive diagnoses. PMID- 8249503 TI - Routine use of endometrial cytology in clinical practice. AB - The aim of this study was to evaluate the long-term use of endometrial cytology with the Endopap sampler in clinical practice. Emphasis was placed on diagnostic accuracy, reduction of the number of curettages and cost per diagnostic test. Blind comparisons of two diagnostic tests were made during an observation period of five years. Endometrial cytology was compared with endometrial curettage as a standard if both samples had been taken within a six-month period and sufficient material had been obtained with both methods. A total of 468 women were studied. All had undergone endometrial curettage because of vaginal bleeding or impending hysterectomy. Abnormal endometrial cytology was found in 129 of 134 (96%) samples from patients with endometrial cancer, in 23 of 25 (92%) with invasive cervical cancer, in 30 of 31 (97%) with atypical endometrial hyperplasia and in 90 of 100 (90%) with endometrial hyperplasia but no atypia. Specificity was 84%. The annual numbers of fractional curettages decreased from 900 in 1986 to 298 in 1991. The study clearly showed that endometrial cytology is useful in clinical practice. PMID- 8249504 TI - Cytology of vulvar squamous neoplasia. AB - Cytologic findings on various vulvar squamous lesions are described in order to elucidate the usefulness of vulvar cytology. Lichen sclerosus, hyperplastic dystrophy and dysplasia with a few exfoliated anucleate squamous cells could not be differentiated cytologically. Numerous parakeratotic cells and dyskaryotic cells were identified in squamous cell carcinoma in situ. Three cytologic patterns were presented in cases of frankly invasive squamous cell carcinoma: negative cytology with parakeratotic cells, suspicious cytology with dyskaryotic cells and positive cytology with malignant cells. Verrucous carcinoma yielded only anucleate squamous cells. Parakeratotic cells without nuclear atypia seemed to be neoplastic cells on vulvar cytology. PMID- 8249505 TI - Cervical cytology after cryosurgery, laser ablation and conization. A comparison of the cotton swab and endocervical brush. AB - Obtaining an adequate Papanicolaou smear, as evidenced by the presence of endocervical cells, is paramount in following patients after treatment for cervical dysplasia. A randomized, prospective trial was done in which 203 patients with a history of cryosurgery, laser ablation or cone biopsy were randomized into two groups: 114 patients had cervical smears done with a cotton swab and 89 patients with an endocervical brush. The presence or absence of endocervical cells was noted on the cytology report. Overall, the endocervical brush produced significantly more adequate smears than did the cotton swab (86% vs. 74%, P = .0389). Within each subgroup the endocervical brush consistently produced a higher number of adequate smears than did the cotton swab: after cryosurgery, 86% vs. 82%, P = .7242; after laser ablation, 95% vs. 88%, P = .61; after conization, 76% vs. 47%, P = .0675. The difference was most notable in the conization group. The endocervical brush is superior to the cotton swab in obtaining satisfactory Papanicolaou smears after cryosurgery, laser ablation and conization. PMID- 8249506 TI - Diagnostic value of differential brushing of all branches of the bronchi in patients with sputum positive or suspected positive for lung cancer. AB - In roentgenographically occult lung cancer, it is often difficult to determine the location of the tumor despite the existence of cancer. This complicates diagnosis and points to a need for a more systematic method of examination. Differential brushing was performed on all the respective segmental bronchi in both lungs of 196 patients with positive or suspected positive indications of lung cancer as revealed by sputum cytology. Fifty-nine borderline lesions in 43 cases and 107 lung cancer lesions in 95 cases were diagnosed. Localization was possible in 70.4% of the cases. The diagnosis of borderline lesions was also possible. At the first examination, the rate of localization, as compared with that in the historical control group, improved from 64.1% to 95.8%, and, in particular, an improvement from 0% to 86.2% was noted in those cases in which abnormal bronchoscopic findings were not observed. Concurrent multiple primary cancer was also diagnosed in 12.6% of lung cancer cases before treatment. With this method, cytologic findings in sputum and in specimens obtained by brushing and histologic findings of resected lung can be compared in an integrated manner, and henceforth more accurate diagnostic criteria can be established. PMID- 8249507 TI - Cytology of atypical and malignant meningiomas in intraoperative crush preparations. AB - Six cases of atypical and malignant meningioma studied by intraoperative crush preparations are reported. Four atypical meningiomas were characterized by large cohesive and dyshesive cell clusters showing slightly pleomorphic nuclei and scant cytoplasm. The papillary meningioma yielded dyshesive groups of pleomorphic cells with variable and well-defined cytoplasm. The malignant meningioma, hemangiopericytic type, showed dyshesive cells with scant, ill-defined cytoplasm and spindle-shaped or vesicular nuclei. PMID- 8249508 TI - Diagnostic accuracy of fine needle aspiration cytology in pancreatic lesions. A review of 77 cases. AB - Fine needle aspiration cytologic specimens from pancreatic lesions in 77 patients were blindly reviewed. The patients were divided into two subgroups: group A included 19 patients without malignancy of the pancreas, and group B included 58 patients with confirmed malignancy (primary or secondary) of the pancreas. The original cytologic diagnoses in both groups were plotted against the respective reevaluated cytologic diagnoses. In group A there were no false-positive cytologic diagnoses, neither originally nor on reevaluation. In group B, among nine cases originally diagnosed as "benign cells," there were two cases diagnosed as malignant on reevaluation, while in the remaining seven cases malignant cells could not be found on reevaluation. In 11 cases in group B the cell material was considered insufficient for cytologic diagnosis, both originally and on reevaluation. In 34 of the 58 cases (58.6%) in group B a conclusive cytologic diagnosis of malignancy was made both in the original cytologic reports and on reevaluation. In this series, sampling errors accounted for most false-negative cytologic diagnoses. The specificity in our series was 100%. The demonstrated sensitivity was 59% initially and 62% on reevaluation. PMID- 8249509 TI - Metastatic cutaneous and subcutaneous deposits from internal carcinoma. An analysis of cases diagnosed by fine needle aspiration. AB - Thirty-eight cases diagnosed by fine needle aspiration as cutaneous/subcutaneous deposits from internal carcinomas were analyzed. Eighteen patients had undergone previous surgery for primary neoplasms in the breast (9 cases), prostate (1), kidney (1), ovary (1), rectum (3), cervix (1), cheek (1) and periampullary region (1). The metastatic deposits in these cases occurred 1-42 months following removal of the primary neoplasm. Sixteen patients had metastatic deposits occurring simultaneously with the primary tumor in the lung (4 cases), pancreas (1), stomach (3), esophagus (1), ovary (1), rectum (1) anal canal (1), maxillary antrum (1), gallbladder (1), colon (1) and kidney (1). In four patients with metastatic nodules, the primary was undetected in spite of a thorough clinical examination and laboratory investigations. There were five cases of Sister Mary Joseph's nodule (umbilical metastases), four from primary cancer in the abdomen and one in which the primary remained unknown. The deposits were solitary except in one case and were usually less than 2.5 cm in diameter. Adnexal tumor can be a differential diagnosis. Special stains were of little help in such cases, and correlation with the clinical findings was enough to arrive at the diagnosis. Fine needle aspiration provides a rapid morphologic diagnosis in patients with cutaneous/subcutaneous metastatic deposits from internal carcinoma. PMID- 8249510 TI - Low grade transitional cell carcinoma and instrument artifact. A challenge in urinary cytology. AB - This was a retrospective study of 46 cases of urine specimens to distinguish low grade transitional cell carcinoma (TCC) and instrument artifact. The cytomorphologic features of 46 urinary specimens showing papillary fragments were studied. Urine from TCC showed papillary and irregular groups of cells with ragged borders and subtle nuclear changes. Urine after instrumentation showed cellular specimens with cohesive, ball-shaped and papillary clusters with smooth borders lined with a denser-staining cytoplasmic collar. The evenness of the border and presence of the cytoplasmic collar were distinguishing features of instrument artifact. PMID- 8249511 TI - Fine needle aspiration biopsy of orbital and eyelid lesions. AB - Thirty-five patients with orbital and eyelid lesions were subjected to fine needle aspiration biopsy (FNAB). Histopathology or other investigations were available in 18 cases; in 17 cases (94%) a concordant diagnosis was reached. There was one false-positive diagnosis in 18 cases (5.5%). FNAB is a reliable and safe diagnostic technique and can be used routinely in outpatient departments for the diagnosis of orbital lesions. It is particularly useful in nonresectable and retrobulbar orbital lesions. PMID- 8249512 TI - Microcystic adenoma of the pancreas. A report of two cases with fine needle aspiration cytology and differential diagnosis. PMID- 8249513 TI - Meningioma, neurilemmoma and astrocytoma specimens obtained with the squash method for cytodiagnosis. A cytologic and immunochemical study. AB - In this study, the peculiar cytomorphologic features of meningiomas and neurilemmomas were described in comparison with those of astrocytomas, employing squash preparations stained by the Papanicolaou method. The value of this method as an adjunct to rapid frozen diagnosis was demonstrated. Meningioma cells had common characteristic features in both cell clusters and individual cells; in the subclass of meningioma they also had peculiar features. Neurilemmomas of type A tissue of Antoni or of type B were characterized by the peculiar shapes of cell clusters and individual cells. In malignant meningiomas the cytologic features were pleomorphic, like those of metastatic carcinoma, making diagnosis difficult. By immunoperoxidase staining for vimentin, the characteristic differences in intracytoplasmic filaments of meningiomas and neurilemmomas could be shown more distinctly. PMID- 8249514 TI - Ploidy studies by image analysis on fine needle aspirates of the breast. AB - Quantitative DNA analysis was performed on preoperative fine needle aspirates of 12 breast cancers and 1 benign lesion using the Cell Analysis Systems (CAS) Model 200 image analysis system. The smears were prepared on CAS slides, routinely processed for Papanicolaou staining, and then destained and restained with Feulgen stain after cytologic evaluation. Four cases were DNA diploid, 3 were tetraploid, 1 was diploid/tetraploid, and 5 were aneuploid. When compared to analysis of samples from the resected tumor, both DNA index and S-phase fraction were constant. PMID- 8249515 TI - Fine needle aspiration for diagnosing a thymoma producing CA-125. A case report. AB - This case report describes the cytologic appearance of a solid and cystic thymoma with high CA-125 content (35,532 microns/mL, measured by enzyme immunoassay) in the cyst fluid obtained by fine needle aspiration (FNA). The initial FNA yielded straw-colored fluid that contained scanty foam cells, lymphocytes and benign epithelial cells and was interpreted as a bronchial inflammatory cyst. The cyst fluid CA-125 was 35,532 microns/mL and serum CA-125, 5.2 microns/mL (normal < 15). In view of the discordance between the cytologic diagnosis, suspicious radiologic appearance and high fluid CA-125 level, FNA of the solid portion of the mass was performed. Smears/cell block examination showed large, cohesive clusters of benign, spindle-shaped epithelial cells (keratin positive) admixed with mature lymphocytes diagnostic of thymoma. This report illustrates the importance of (1) sampling solid areas of solid/cystic tumors, (2) rapid assessment of FNA material, and (3) appropriate clinical and radiologic correlation. It also describes the cytologic features of a solid/cystic thymoma with elevated CA-125 content in the cyst fluid. PMID- 8249516 TI - Fine needle aspiration biopsy cytology of sclerosing hemangioma of the lung. A case report. AB - We report the fine needle aspiration cytology of a case of sclerosing hemangioma of the lung. This rare and benign neoplasm characteristically exhibits a variety of histologic patterns in a particular tumor--e.g., angiomatous, solid, papillary and sclerotic. There are few descriptions in the cytologic literature, and all are of the angiomatous pattern. Our case showed a papillary/sclerotic pattern with no angiomatous area. The cytologic findings consisted of spindle cells admixed with sheets of cuboidal cells, precisely reflecting the histologic pattern. PMID- 8249517 TI - Metastatic glioblastoma multiforme in cervical lymph nodes. Report of a case with diagnosis by fine needle aspiration. AB - Metastatic glioblastoma multiforme is a rare event usually seen in patients with previous and repeated surgery. Fine needle aspiration from enlarged lymph nodes in a patient with cerebral glioblastoma multiforme showed a highly cellular smear with dispersed single cells and cell clusters. The cells had marked pleomorphism, hyperchromasia, coarse chromatin, prominent nucleoli and occasional intranuclear cytoplasmic inclusions. Multinucleate giant cells and marked endothelial proliferation were also seen. The diagnosis of metastatic glioblastoma multiforme was supported by immunohistochemistry, which showed many cells with thick and large cytoplasmic processes with intense immunoreactivity to glial fibrillary acid protein. The differential diagnosis with other metastatic small cell tumors is discussed. PMID- 8249518 TI - Metastatic signet ring carcinoma of the breast as a source of positive cervicovaginal cytology. A case report. AB - The cellular manifestations of extrauterine cancer have been well documented. While finding metastatic carcinoma in gynecologic specimens should not be unexpected, descriptions of certain types of neoplasms are not well represented in the cytology literature. We describe the clinical, cytopathologic and histologic features of a case of signet ring cell breast carcinoma metastatic to the uterine cervix. Signet ring cell breast carcinoma is a rare neoplasm, and its possible histogenesis is discussed. PMID- 8249519 TI - Fine needle aspiration biopsy of medullary carcinoma of the thyroid metastatic to the liver. A case report. AB - A 59-year-old female with a six-month history of medullary carcinoma of the thyroid (MCT) presented with multiple liver nodules and several physical complaints. Ultrasound-guided fine needle aspiration biopsy (FNAB) of the liver established the diagnosis of metastatic MCT. This report describes the usefulness of FNAB, aided by immunocytochemical and ultrastructural studies, in patients with MCT in whom liver metastases are suspected. PMID- 8249520 TI - Nodular fasciitis of the orbit diagnosed by fine needle aspiration cytology. A case report. AB - Nodular fasciitis is a common lesion in the head and neck area in the pediatric population. In adults the extremities are the favored location. A 10-year-old girl developed a rapidly growing orbital mass that resulted in medial deviation of the eyeglobe. Clinical diagnosis was a malignant neoplasm, probably rhabdomyosarcoma. A fine needle aspiration biopsy yielded a preoperative diagnosis of nodular fasciitis. The patient underwent surgical resection and had histologic confirmation of the cytologic diagnosis. PMID- 8249521 TI - Squamous cell carcinoma of the breast. Report of a case diagnosed by fine needle aspiration cytology. AB - A 66-year-old woman presented with pure squamous cell carcinoma of the breast, a rare occurrence. The incidence of pure squamous cell carcinoma of the breast is lower than that of the mixed type. The preoperative diagnosis could be made by fine needle aspiration cytology. A modified radical mastectomy was performed. The patient was doing well, with no evidence of recurrence, 17 months after surgery. PMID- 8249522 TI - Biliary cystadenocarcinoma arising in a cystadenoma. Report of a case diagnosed by fine needle aspiration cytology. AB - Hepatic cyst fluid cytology tends to yield disappointing results. We report a case of a 56-year-old woman with a biliary cystadenocarcinoma diagnosed by fine needle aspiration cytology. Computed tomography scans had shown a solitary, unilocular hepatic cyst over a five-year period. There was a recent increase in the size and development of a mural echogenic focus. Cytologic examination revealed clusters of malignant glandular cells in a background of cellular debris and mucinophages. The resected specimen confirmed the presence of an adenocarcinoma arising from malignant transformation of a preexisting cystadenoma. PMID- 8249523 TI - Fine needle aspiration cytodiagnosis of basaloid-squamous cell carcinoma metastatic to a cervical lymph node. PMID- 8249524 TI - Statistics and cervical sampling methods. PMID- 8249525 TI - Retroperitoneal human dirofilariasis with a pseudotumoral presentation: comments. PMID- 8249526 TI - [Gastroduodenal mucosa lesions. New physiopathological concepts]. PMID- 8249527 TI - [Idiopathic acute pancreatitis (IAP)]. PMID- 8249528 TI - [Gastric hypersecretion, Helicobacter pylori, and duodenal ulcer]. PMID- 8249529 TI - [Molsidomine and nitric oxide in gastric cytoprotection]. AB - The effect of Molsidomine as a gastric mucosa protector against absolute ethanol was studied in Wistar rats. It was found that Molsidomine provided an important gastric cytoprotection and it behaved in a similar way as the nitric oxide (NO), the latter verified by specific NO antagonist, namely the NG-monomethyl-L arginine. It was also demonstrated that the cytoprotector mechanism of Molsidomine, as well as of other nitroderivates was sulfhydryl and HEM dependent regarding the guanylate-cyclase enzyme. It can be concluded that both Molsidomine and the NO behaved as modulator of the gastric microcirculation, venous and arterial vasodilator and that their gastric cytoprotective mechanism was endogenous prostaglandin dependent. PMID- 8249530 TI - [Increase of acetaminophen conjugation ability in experimental cholestasis]. AB - Acetaminophen glucuronidation ability and its relationship to hepatic microsomal phospholipid changes were studied during experimental extrahepatic cholestasis. This study included two groups of Wistar rats: 1) normal rats and 2) cholestatic rats. UDP-Glucuronyltransferase activity (UDP-GT) towards acetaminophen resulted increased in 38% (p < 0.05) in the cholestatic group when compared to normal rats. We also found important changes in hepatocyte microsomal phospholipid profile. When phosphatidylcholine fatty acid composition was analysed, all of them resulted diminished except for an increment on oleic acid content (36%). This fatty acid increase in the main membrane phospholipid had a parallel behaviour with acetaminophen glucuronidation activity. This fact could support an activation role for oleic acid over this enzymic system. PMID- 8249531 TI - HBV and HCV infection in kidney transplant recipients. AB - The risk of HBV and HCV liver infection in kidney graft recipients was evaluated in 35 patients. All were tested for anti-HBc, HBsAg, HBeAg, anti-HBs, Anti-HBe, anti-HCV (c-100-3 and c-100-3, c-22, 33-c), anti-HDV and anti-HIV by ELISA, and for HBV-DNA by hybridization. Liver biopsy, immunostaining for HBcAg and Knodell's hepatic inflammatory index were performed in 18. Mean time elapsing form transplant to inclusion was 20.7 months (range 1-108). HBsAg was the only marker searched for prior to transplant. Twenty six (74.2%) patients presented HBV and/or HCV markers, while 9 (25.8%) had none; 16 (45%) proved anti-HBc+, 6(17.1%) HBsAg+, (3 HBeAg+ and 3 anti-HBe+), 7 (20%) anti-HBs+ and 3 (8.5%) isolated anti-HBc. Anti-HCV (C-100-3) was positive in 9/32 (28.1%), while 2nd. generation anti-HCV was positive in 20/35 (57.1%) cases. No false positives for 1st. generation test were found. Both anti-HDV and anti-HIV were negative in all the sample. Raised aminotransferases were present in 13/30 (43.3%), 7 in anti HCV+, one in HBsAg+ and 3 in HBsAg+/HCV+ cases, but normal in 17/30 (56.6%). History of Transfusion and Hemodialysis time showed no significant differences between anti-HCV+ and anti-HCV negative cases. Biopsy disclosed 10 chronic persistent hepatitis (CPH), one chronic active hepatitis (CAH) with cirrhosis, one inactive cirrhosis (Ci) 4 minimal lesions (MHL) and 2 normal. Seven CPH, 3 MHL. one normal and both cirrhosis cases proved anti-HCV+. HBsAg was positive in the single CAH, in 2 CPH and in one MHL.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249532 TI - [Mitotic index of intraepithelial lymphocytes in children with celiac disease. Retrospective study]. AB - The mitotic activity of epithelial lymphocytes was determined in 69 jejunal biopsies performed in 23 children with celiac disease. The mitotic index (MI) exceeded 0.2% in 46 biopsies obtained in untreated celiac and during acute gluten challenge. In the 23 biopsies made during the period of gluten restriction the mitotic index fell down under 0.2%. This retrospective study confirm the reproducibility of the method proposed by Marsh. The high mitotic index in the first biopsy may be useful to reduce in the future the multiple biopsies currently necessary for the diagnosing celiac disease. PMID- 8249533 TI - [Flumazenil, a benzodiazepine antagonist, in endoscopic premedication]. AB - Midazolam is the benzodiazepine recommended for anaesthesia induction when an early recovering is desirable. Flumazenil, which is also a hydrosoluble imidazobenzodiazepine, improves the safety of the forhere one because it immediately reduces the degree of sedation. With this purpose, this last drug has been studied in 25 patients who had been previously treated with Midazolam in order to carry out different endoscopic studies. The Flumazenil average dose used was 0.276 mg and the conscience recovering after its intravenous injection, evaluated by a simple oral numeric regression test, oscillated between 60 and 300 seconds with an X of 96 seconds. Once the study was finished, all patients were discharged from hospital after two-hour observation, except for one patient (4%) who suffered a complication and so was discharged after a twenty-four-hour control. No doubt the advent of a drug opposed to benzodiazepine receptors used in anaesthetic induction shows the possibility of a quick recovering and moreover. Quicker discharge from hospital. PMID- 8249534 TI - [Esophageal cancer associated to a middle esophageal diverticulum: a case report]. PMID- 8249535 TI - [Erythro-tropic human parvovirus B19 (HPV)]. PMID- 8249536 TI - Genetic lesions in preleukemia. AB - The incidence of genetic abnormalities have been investigated in a variety of preleukaemic states RAS and FMS oncogene, p53 suppressor gene mutations and monoclonality in myelodysplastic syndromes (MDS), a paradigm for pre-leukemias have been observed. Other patients at risk of developing either secondary leukaemia or evolving into leukaemia have been similarly studied including haematologically normal patients in remission from lymphoma. Time from treatment to detection of genetic abnormalities is a significant factor in some of these patients which is consistent with the expansion of an abnormal clone. A case of non-dysplastic MDS has been identified with a 7q-karyotypic abnormality typical of therapy related MDS, abnormal progenitor growth and RAS mutations but with normal clinical features. Normal individuals have also been under investigation and found to have a low incidence of proto-oncogene mutations. A prospective study should enable us to determine if these parameters are indeed prognostic indicators. PMID- 8249537 TI - [Chronic myeloid leukemia without Philadelphia chromosome (Ph-negative CML)]. AB - Ph-negative chronic myeloid leukemia [Ph(-)CML] is a heterogenous group of conditions characterized by similar cytogenetic pattern but variable changes at the molecular level. All cases with BCR gene rearrangement in this group have clinical and haematological course similar to Ph(+)CML. However, CML without rearrangements of ABL and BCR genes, called "atypical CML", forms a separate entity showing clinical and morphological features different from classical CML. PMID- 8249538 TI - [Diagnosis and treatment of central venous catheter related infections]. AB - The central venous catheter related infections are relatively common in clinical practice. These infections are particularly frequent in the patients with long term central venous catheter, such as the oncology patients or patients with short intestine syndrome. The diagnosis of infection should be based on repeated positive blood cultures. In some cases the infection responds well to antibiotic treatment. Unless there are indications for removal of catheter antibiotic therapy should be applied for 1-2 weeks. PMID- 8249539 TI - [Platelets and the fibrinolytic system]. AB - The authors present interactions between fibrinolysis and platelets. Platelets may exert both stimulatory and inhibitory effect on fibrinolysis. Also the problem of fibrinolysis activation on platelets function is considered. PMID- 8249540 TI - Standard heparin and low molecular weight heparin effect on spontaneous platelet aggregation measured in whole blood and in platelet-rich plasma. AB - The evaluation of both standard heparin (SH) and low molecular weight heparin (LMWH) (Fraxiparine) impact on spontaneous platelet aggregation assessed in the whole blood and in platelet-rich plasma (PRP). The results obtained reveal that SH enhance spontaneous aggregation in a similar degree both in the whole blood (34.5 +/- 3.0, control 21.3 +/- 4.1) and in PRP (33.5 +/- 2.7, control 13.8 +/- 3.2). LMWH significantly intensifies spontaneous aggregation in the whole blood (46.9 +/- 3.2, control 21.3 +/- 4.1) but not in PRP (19.4 +/- 4.3, control 13.8 +/- 3.2). PMID- 8249541 TI - [Diagnostic value of acidified glycerol lysis test (AGLT) in hereditary spherocytosis and selected hematologic diseases]. AB - AGLT and osmotic fragility test (method according to Dacie) were performed in patients with hereditary spherocytosis, their relatives and in patients with other hematological diseases. Abnormal results of both tests were observed not only in all cases of hereditary spherocytosis but also in other patients with spherocytes present in the peripheral blood film. In our opinion, AGLT is a good screening test for hereditary spherocytosis and can replace the more time consuming Dacie's method. PMID- 8249542 TI - Dr. Jean Watson. Director: Center for Human Caring Denver, Colorado. Interview by Eve Henderson. PMID- 8249543 TI - Placebos: trickery or therapy? PMID- 8249545 TI - Alternate models for direct access to nursing services provided by registered nurses. PMID- 8249544 TI - Advance treatment directives: a nurse ethicist's view. PMID- 8249546 TI - Determining the appropriate scope of nursing practice for particular practice settings: guidelines for registered nurses: Alberta Association of Registered Nurses. AB - The depth and breadth of the scope of nursing practice in Alberta places registered nurses in a position where they make a significant contribution to health care. The nature of the contribution of registered nurses in a given practice setting must rely on a determination of the appropriate scope of practice for registered nurses in that setting. That determination should be guided by the principles outlined above, and in accordance with the documents cited in this paper. Within these guiding principles, activities performed by registered nurses in a given practice setting should be mutually agreed upon between health care professionals, should be time constant, and should be driven by the needs and consent of the client, not by the desire for convenience in health care professionals in the practice setting. PMID- 8249548 TI - Supportive environments: a conceptual model. PMID- 8249547 TI - Will I become addicted? PMID- 8249549 TI - The lowly louse. PMID- 8249550 TI - Painful breastfeeding. PMID- 8249551 TI - Pharmacokinetics and related pharmacodynamics of anticholinergic drugs. AB - The pharmacokinetics and some pharmacodynamic properties of atropine, glycopyrrolate and scopolamine are reviewed. With the development of new analytical methods for drug determination, it is now possible to measure relatively low concentrations of these drugs in biological fluids and, consequently, some new kinetic data have been collected. Following intravenous administration, a fast disappearance from the circulation is observed and due to a high total clearance value their elimination phase half-lives vary from 1 to 4 h. All these agents are nonselective muscarinic receptor antagonists, but their actions on various organ systems with cholinergic innervation show considerable diversity. The cardiovascular effects are of short duration; other peripheral muscarinic effects and CNS effects can last up to 8 h or even longer. Differing from atropine and scopolamine, glycopyrrolate as a quaternary amine penetrates the biological membranes (blood-CNS, placental barriers) slowly and incompletely, making it the drug of choice for elderly patients with coexisting diseases and for obstetric use. Similarly, its oral absorption is slow and erratic, and hence it cannot be used as an oral premedicant. Atropine, scopolamine and glycopyrrolate have a definitely faster absorption rate, when injected into the deltoid muscle compared with administration into the gluteal or vastus lateralis muscles. There appear to be significant differences in the metabolism and renal excretion of these agents. Scopolamine is apparently excreted into the urine mainly as inactive metabolites, nearly half of the atropine dose administered is recovered in the urine as the parent drug or as active metabolites and about 80% of glycopyrrolate is excreted as unchanged drug or active metabolites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249552 TI - Persistent postoperative hiccups: a review. AB - The pathogenesis of persistent postoperative hiccups is not known. Hiccups can present as a symptom of a subphrenic abscess of gastric distention, and metabolic alterations may also cause hiccups. The hiccups may develop because of increased activity in neural reflex pathways not yet fully defined. Numerous treatment modalities have been tried but with questionable success. Valproate has proven effective in two trials investigating persistent non-surgical hiccups. The simple application of a nasogastric tube may successfully treat the hiccups, possibly because of an alteration of the activity in the reflex neural pathways involved. The available literature on the treatment of persistent hiccups is reviewed, and a treatment protocol for persistent postoperative hiccups is provided. PMID- 8249553 TI - Halothane-associated enhancement of the secondary immune response to sheep erythrocytes in mice: cell transfer studies. AB - The effect of halothane anesthesia on the humoral immune response to sheep red blood cells was studied in mice immunized twice, with a 15-day interval. On both occasions, mice were exposed to 1.5% halothane for 40 min immediately after sensitization. Halothane reexposure resulted in increased numbers of IgG secreting cells (IgG-SC) as well as circulating 7S-serum agglutinins. To examine further whether this effect could be obtained in syngeneic recipients, adoptive transfer experiments employing spleen cells were performed. While mice receiving cells from unimmunized and anesthetized donors displayed significantly higher levels of IgG-SC, recipients of cells from normal, immunized and immunized anesthetized donors showed a depressed response when compared to control counterparts. Besides the possibility of an enhancing effect of halothane reexposure on the humoral response, this procedure may counteract normal physiological immunoregulatory processes during the generation of the immune response. PMID- 8249554 TI - Patient-controlled analgesia (PCA) leads to more postoperative pain relief, but also to more fatigue and less vigour. AB - This investigation evaluated patient-controlled analgesia (PCA) for subjective well-being and mood in the postoperative period in comparison with the intramuscular (im) administration of morphine given on demand. Patients scheduled for elective upper abdominal surgery were assigned at random to either PCA (n = 17) or im morphine (n = 14). The PCA group experienced significantly more pain relief and consumed more morphine than those who received im morphine. The PCA patients suffered from more fatigue and showed less vigour than the im group. Neither preoperative trait anxiety nor locus of control was associated with postoperative pain in either of the groups. PMID- 8249555 TI - Epidural morphine for postoperative pain relief in children. AB - Epidural morphine for postoperative pain relief is in general use, and has proved to be very efficient in adults. The epidural technique and the use of epidural morphine are much less frequent in children. For 2 years we have prospectively followed 76 children who had epidural morphine for postoperative pain relief after major abdominal surgery. The age distribution was from newborn to 13 years, with a median age of 12 months. It was estimated that 94% of the patients had good analgesia for the first 24 postoperative hours and no other opioids were given. The side effects were few, but one case of respiratory depression was seen and 20% of the children had pruritus. There were four dural punctures and three catheters slipped out accidentally, but otherwise the treatment was continued as long as it was considered necessary (1-11 days). The use of postoperative ventilatory support decreased during the investigation. We observed a change in the sleeping pattern with an increased number of sleep-induced myoclonia during the administration of epidural morphine. In conclusion, the use of epidural morphine in children for postoperative pain relief is very efficient. The minimal effective dose has not been established as yet, but 50 micrograms/kg every 8 h, supplemented with small doses of bupivacaine, provides excellent analgesia in the immediate postoperative period after major abdominal surgery. The side effects are few, but the risk of respiratory depression is always present and observation in the intensive care unit or recovery for the first 24 h is strongly recommended. PMID- 8249556 TI - Attenuation of the pressor response to tracheal intubation with oral nitrendipine. AB - The effect of nitrendipine on the cardiovascular responses to tracheal intubation was studied in a placebo-controlled, randomised, double-blind trial. Thirty patients (ASA physical status 1) undergoing elective surgery received either 5 or 10 mg nitrendipine, or a placebo orally 3 h before induction of anaesthesia (n = 10 for each group). Anaesthesia was induced with sodium thiopentone 5 mg/kg i.v. and tracheal intubation was facilitated with vecuronium 0.2 mg/kg i.v. Patients receiving the placebo showed a significant increase in the mean arterial pressure and the rate-pressure product in response to tracheal intubation. These increases following intubation were reduced in nitrendipine-treated patients compared with the placebo group (P < 0.05). Oral administration of nitrendipine (5 or 10 mg, 3 h before induction of anaesthesia) was able to attenuate the hypertensive response to tracheal intubation in ASA 1 patients under light anaesthesia. We propose this pharmacological technique with supplementary doses of opioids and/or benzodiazepines for the management of patients with hypertension or coronary artery disease. PMID- 8249557 TI - The independent metabolic effects of halothane and isoflurane anaesthesia. AB - Twelve healthy, unpremedicated women scheduled for total abdominal hysterectomy were given either isoflurane (n = 6) or halothane (n = 6) anaesthesia. They all received general anaesthesia for a period of 3 h, with surgery being carried out only in the last hour. The anaesthesia consisted of thiopentone, pancuronium and a mixture of oxygen-enriched air (FiO2 = 34%) supplemented with 1 MAC of either isoflurane or halothane. The patients were maintained normothermic, and with an arterial SaO2 above 95% throughout the period of the study. The following measurements were made before, during and after anaesthesia (with and without surgery): oxygen consumption (VO2), carbon dioxide production (VCO2); circulating concentrations of various hormones (insulin, growth hormone and cortisol); various metabolites; selected amino acids and albumin; forearm arterio-venous concentration difference of glucose, lactate, free fatty-acids and selected amino acids (four patients in each group). Whole body VO2 decreased significantly by over 20% during anaesthesia (with or without surgery), P < 0.05). Although the circulating concentration of most amino acids showed little or no change during anaesthesia alone, there was a tendency for the flux of most metabolites to decrease, and this persisted during surgery (P < 0.05). During anaesthesia alone there was a twofold reduction in the plasma cortisol concentration (P < 0.05), and a decrease in albumin concentration (P < 0.01). With the onset of surgery, plasma cortisol concentration increased rapidly (in association with several other hormones and metabolites) but hypoalbuminemia persisted. PMID- 8249558 TI - Peroperative nitrous oxide does not influence recovery after laparoscopic cholecystectomy. AB - We have evaluated the effects of nitrous oxide on recovery following laparoscopic cholecystectomy in a prospective, randomised, double-blind study with 42 otherwise healthy patients. All patients received meperidine 1 mg/kg and atropine 6 micrograms/kg im for premedication, and anaesthesia was induced with fentanyl 2 micrograms/kg and thiopental 4-6 mg/kg. Succinylcholine was used for the intubation and muscle relaxation was achieved using vecuronium. Isoflurane with 70% nitrous oxide in oxygen and fentanyl was used for maintenance of anaesthesia in group I (n = 19), and isoflurane in air/oxygen and fentanyl in group II (n = 23). The postoperative ward staff and the surgeon evaluating the postoperative recovery were blinded to the anaesthetic technique. No differences were found in duration of operation and anaesthesia, need for postoperative analgesia or postoperative nausea treated medically. Recovery, judged by the Steward Coma Score, comprehension and collaboration, degree of sedation and orientation in time and space, was similar in the two groups. Postoperative hospital stay was 1 (1-4) day in the nitrous oxide group (median (10-90th percentiles) versus 2 (1-4) days in the air group. The time until patients were recovered, as judged by return to work and normal daily activities, was the same in the two groups: 8 (4 11) days in the nitrous oxide group and 8 (4-11) days in the air group. We conclude that nitrous oxide has no influence on recovery after laparoscopic cholecystectomy. PMID- 8249559 TI - Evaluation of a new system for ventilatory administration of nitric oxide. AB - A new system for delivery of nitric oxide (NO) to inspiratory gas consisting of two mass flow regulators and a soda-lime absorber for scavenging of nitrogen dioxide (NO2) is described. The system was evaluated using three different techniques for NO analysis (infrared, chemiluminescence and electro-chemical fuel cell technique). The electro-chemical fuel cell was less sensitive to humidity in the sample and is suitable for clinical routine use. The infrared analyser was very sensitive to humidity and the gas sample must be dried by silica gel, which absorbs NO2 and will cause falsely low NO2 values. NO2 was analysed with ultra violet methodology. NO2 is highly toxic and the highest recommended occupational health and safety level for inhalation is 5 ppm. The highest values of NO2 in our system were detected before the absorber in the inspiratory limb of the breathing system, being 5 ppm at 100% oxygen and 100 ppm NO using "infant" respiratory settings (3 l/min in ventilation, frequency of 30/min). The corresponding value for "adult" respiratory settings (10 l/min in ventilation, frequency of 15/min) was 3.2 ppm. The absorber reduced these levels to well below 1 ppm. When clinically relevant levels of NO were used (20 ppm), no NO2 could be detected after the absorber, irrespective of oxygen concentration in the breathing gas. It was observed that gas cylinders with NO mixed in nitrogen may initially have a high NO2 concentration (around 12 ppm) and should be flushed thoroughly before use. PMID- 8249560 TI - The influence of nitrous oxide on recovery of bowel function after abdominal hysterectomy. AB - The influence of nitrous oxide on the recovery of bowel function was studied in 36 patients anaesthetised for elective abdominal hysterectomy with or without salpingo-oophorectomy. Patients were randomly assigned to receive either isoflurane in nitrous oxide and 30% oxygen (N2O group) or isoflurane in air and 30% oxygen (Air group). Anaesthetic management included thiopentone, fentanyl, suxamethonium and atracurium. The lungs were not ventilated prior to intubation. Before closing the abdomen, the surgeon assessed the degree of distension of the intestines and the closing conditions. Postoperative nausea and vomiting was assessed 2, 6, 12 and 24 h after recovery from anaesthesia. The lapse of time before mobilisation and passing of flatus and faeces was recorded. The patients in the Air group were significantly older than the patients in the N2O group (48.9 years versus 44.0 years, P = 0.04); otherwise, there were no differences in the demographic data of the patients. We found no significant differences between the groups with respect to nausea and vomiting, distension of the intestines before closure of the abdomen, closing conditions, time elapsing before mobilisation, constipation before recovery of bowel function or time elapsing before passing of flatus. We found a statistically significant delay of 10.3 h in time elapsing before passing of faeces in the N2O group compared to the Air group (P = 0.04), suggesting a potentially adverse effect of nitrous oxide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249561 TI - Comparison of 5 mg tetracaine diluted in 1 ml, 2 ml and 4 ml of 10% glucose for spinal anesthesia. AB - The influence of variations in the volume of injectate when maintaining an identical dose of a hyperbaric local anesthetic has not previously been investigated when performing spinal anesthesia. This study compares spinal anesthesia of 5 mg of lyophilized tetracaine diluted in 1 ml, 2 ml or 4 ml of 10% glucose in 45 elderly patients undergoing elective hip surgery. While supine and horizontal, each patient received double-blind one of the three solutions through a catheter inserted 4 cm into the intrathecal space at the L2-L3 or L3-L4 interspace. No difference in the anesthetic effects was found between the three groups. The median value of the maximal sensory level was T6 (range T3-L2), T4 (range T3-T9) and T5 (range T3-T11) in the 1-ml, 2-ml, and 4-ml groups, respectively. The number of patients with a motor blockade of grade 2 or 3 was 12/15, 14/15 and 13/15, and the time from the initial dose to the need for the first top-up dose (mean +/- s.d.) was 88 +/- 35 min, 75 +/- 15 min and 68 +/- 15 min for the 1-ml, 2-ml and 4-ml groups, respectively. Hemodynamic changes were also comparable between the three groups. The authors conclude that in elderly patients, undergoing spinal anesthesia while supine and horizontal, variations in volume from 1 to 4 ml do not influence the characteristics of hyperbaric spinal anesthesia while injecting an identical dose of local anesthetic. PMID- 8249562 TI - Efficacy of epidural blood patch for postdural puncture headache. AB - This prospective investigation was conducted to evaluate the efficacy of different volumes of epidural blood patch (EBP) for treatment of postdural puncture headache (PDPH) in 81 consecutive patients. In the first part of the investigation (Study part I), 10 ml of blood was injected for EBP in 28 patients. In the second randomized part of the investigation (Study part II), the patients were allocated to receive for EBP either 10 ml (27 patients) or 10-15 ml (26 patients), according to the height of the patient. The procedure was considered initially successful if PDPH disappeared completely during the 2-h recovery room follow-up. To evaluate the long-term success, a questionnaire was mailed to all patients. The EBP performed 3.7 +/- 2.9 days following the dural puncture was initially successful in 88-96% of the patients in the different study groups. In the questionnaire, only 50-68% of the patients reported that PDPH had disappeared immediately without recurrence. In 16-36% of the patients the PDPH returned at lesser intensity and in 14-17% PDPH was reported to have continued, disappearing gradually in all patients. Despite this, 87% of all patients were satisfied with the EBP treatment. There were no statistically significant differences between the groups. The results indicate that a larger, height-adjusted volume of blood for EBP in adults does not produce a better effect on PDPH compared to a standard 10-ml volume. Despite the excellent initial effect (91%) seen in our patients, a permanent effect of the blood patch was only achieved in 61%. PMID- 8249563 TI - Peripheral antinociceptive effects of morphine after burn injury. AB - In a double-blind study, 2 mg of morphine in saline, or saline only, was given subcutaneously into a second-degree bilateral leg-burn injury in 12 volunteers. Heat-pain thresholds and pressure-pain thresholds were significantly increased by local morphine administration. These results confirm experimental data demonstrating a peripheral antinociceptive effects of opioids in inflamed tissue. PMID- 8249564 TI - A short fasting interval does not increase preoperative plasma atrial natriuretic polypeptide concentrations in children. PMID- 8249565 TI - Diversity and developmental regulation of extraocular muscle: progress and prospects. AB - The developmental regulation of skeletal muscle phenotypes has attracted much attention from cell and molecular biologists. Myogenesis serves as an excellent model of the interaction of genetic and epigenetic events in determining definitive adult characteristics. The muscles that are responsible for eye movements are among the most structurally and functionally diverse mammalian skeletal muscles. Despite these unique attributes, the extraocular muscle has been studied less extensively than the traditional limb and diaphragm models. This review explores current concepts regarding the development of these muscles. Proper interocular alignment and coordination of eye movements are essential for normal vision in the adult. Moreover, coordinated oculomotor function during the perinatal period is critical for the normal development of the visual system. Therefore, a thorough understanding of the sequence of events and mechanisms that regulate extraocular muscle development and maldevelopment is vital. Studies related to myoblast origin and differentiation and to the role of genetic versus environmental factors in shaping and maintaining adult extraocular muscle phenotype are reviewed. Prospects for understanding the cellular and molecular mechanisms that regulate the heterogeneity and plasticity of these muscles also are discussed. PMID- 8249566 TI - Hippocampal p75 nerve growth factor receptor immunoreactivity in development, normal aging and senescence. AB - Using the monoclonal antibody ME 20.4, p75 nerve growth factor (NGF) receptor immunoreactivity has been studied in the hippocampus and adjacent cortex in a series of 57 cases ranging in age from 24 weeks gestation to 95 years of age. The activity of the neurotransmitter-synthesizing enzyme choline acetyltransferase (ChAT), the activity of which is regulated by NGF, has also been determined in parallel experiments. p75 NGF receptor immunoreactivity was detected in the fetal neocortex as nerve terminal staining, potentially derived from basal forebrain neurons which were positive for NGF receptor, and was also localized in nerve cells of the cerebral cortex. Cortical reactivity for NGF receptor increased with age up to the 4th decade thereafter remaining constant. NGF receptor reactivity localized to neocortical neuronal cell bodies was not present in the postnatal or adult brain. Hippocampal reactivity for the NGF receptor was not present before birth appearing first in the postnatal period and thereafter showing an identical development pattern to the neocortex. ChAT activity in the entorhinal cortex and hippocampus partially paralleled NGF receptor development being present in the neocortex in the fetus but not in the fetal hippocampal formation and increasing postnatally to reach maximum levels in the 4th decade. Whilst entorhinal cortex ChAT values remain relatively constant with ageing, hippocampal ChAT declined with age after the 4th decade. The results may have implications for the aetiology of age-related cholinergic deficits in the hippocampus. PMID- 8249567 TI - Morphological evidence for migration of particle-laden macrophages through the interalveolar pores of Kohn in the murine lung. AB - We have investigated the topography of particle-laden macrophages in the pulmonary tissue of CD-1 mice after intratracheal instillation of a suspension of 250 micrograms of calcium tungstate. The mice were sacrificed 1, 3, 7 and 14 days after the particle deposition. Lung fragments were studied by scanning electron microscopy (SEM) coupled with X-ray microanalysis that allowed in situ elemental identification of tungsten in the lungs. Tungsten-positive macrophages were distinctly located in the lungs of mice sacrificed at 1-3 days when compared with samples from mice killed 7-14 days after the calcium tungstate instillation. At 1 3 days, the tungsten-carrying macrophages were accumulated near the terminal bronchioles whereas they were seen predominantly in the alveolar ducts and sacs in the 7- to 14-day groups of mice. This suggests that during pulmonary inflammation there is a redistribution of the particle-containing macrophages throughout the deep lung tissue. In high-magnification SEM views, we observed that the tungsten-positive macrophages presented numerous surface microvilli. Tungsten-laden phagocytes were detected in interalveolar fenestrae, at the so called Kohn pores. This finding documents that the Kohn pores may be used by inflammatory cells as a pathway for the migration of phagocytes in between adjacent alveolar sacs. PMID- 8249568 TI - Histotopographic and ultrastructural study on the lymphatic network of the pancreas in the guinea pig. AB - The localization of absorbing lymph vessels was studied in the normal guinea pig pancreas by light microscopy (serial semithin sections) and transmission electron microscopy (ultrathin sections). Absorbing lymph vessels were observed both in the interlobular and intralobular areas where they can be localized close to the adenomeres. Lymph vessels were also seen in the interlobular areas located next to the islets of Langerhans and to groups of endocrine cells (n = 8-10) not showing a vascular organization comparable with islets of Langerhans. All lymph vessels in the pancreas are absorbing lymph vessels, characterized by a very thin endothelial wall, anchoring filaments and the absence of a definitive basal membrane. Adjacent endothelial cells form end-to-end, overlapping and interdigitating intercellular junctions. Valves were present both in the interlobular and intralobular lymph vessels. PMID- 8249569 TI - A morphological study of the size of the vascular compartment of the carotid body in a non-human primate (Cercopithecus ethiopus), and a comparison with the cat and rat. AB - The carotid bodies from 5 adult non-human primates (mean body weight 2.9 kg) were perfusion-fixed at normal arterial blood pressure with 3% phosphate-buffered glutaraldehyde. Serial 5-microns sections were cut, stained, and, using an interactive image analysis system, determinations were made of the volumes of the carotid body and of its vascular and extravascular compartments. The total volume of the carotid body was, on average 0.21 mm3, the total vascular volume contributing 9.7%. The small vessels (5-12 microns diameter) comprised 5.4% of the total volume of the carotid body, or about 56% of the vascular compartment; these estimates were similar to values obtained for the cat and rat. The mean small vessel endothelial area, per unit of extravascular volume (which is assumed to consist largely of type 1 and 2 cells) was 61.8 mm-1 in the primate and 69.7 mm-1 in the cat. A value was not available for the rat. Estimates of the carotid body tissue specific blood flow were 31, 61 and 104 ml/min/100 g organ tissue in the primate, cat and rat, respectively. It was emphasised that these values were not to be confused with estimates of carotid body specific blood flow based on values for total organ blood flow and the dissected weight of the organ. PMID- 8249570 TI - Local vascular contribution of the superficial palmar arch. AB - The aim of the present study was to establish a precise topography of the arterial supply of the different layers in the palm of the human hand. Two hundred upper extremities from fresh human cadavers aged 20-80 years were injected either by coloured latex or by India ink and gelatine. Some of our specimens were treated by the Spalteholz technique of transclarification, while some others were treated by acid corrosion. All of the specimens were dissected under the dissecting microscope. The superficial palmar arch (SPA) is the main vascular structure of the palm of the hand. Regardless of its size and of its variation, in addition to the common palmar digital arteries, the SPA supplies the superficial flexor tendons, flexor retinaculum, median and ulnar nerves, tendon of the flexor pollicis longus muscle, lumbrical muscles, palmar aponeurosis and the skin of the palm of the hand. These data may be of interest to those involved in hand surgery. PMID- 8249571 TI - Arguments for a vascular glaucoma etiology. AB - A material of Open Angle Glaucoma (G) collected by an eye practitioner (B S) from 1980 to 1991 is analysed with special respect to the occurrence of disc haemorrhages (Hs) and retinal vein occlusions (VOs). There is a close association between Hs and VOs and G, which in all categories increases with increasing follow-up time. Furthermore, the occurrence of mixed cases--H in one eye and VO in the same or the other--support the opinion that Hs, branch vein occlusions and central vein occlusions are manifestations of the same vascular disease, the difference between them being the size of the vessel affected. The well known morphological changes in the retinal veins of glaucomas and in central vein occlusions are endothelial proliferations causing progressive increase of flow resistance. Similar changes most probably also cause branch occlusions and disc haemorrhages. The vascular affection which obstructs the vessels with hindrance of the blood flow and impaired nutrition of neuronal tissue, might be the primary cause of glaucoma. PMID- 8249572 TI - Perimetric point density and detection of glaucomatous visual field loss. AB - Eighty-three patients with high risk ocular hypertension were followed between 9 and 120 months with computerized threshold perimetry every three months in a prospective clinical study. Twenty eyes developed definite glaucomatous visual field defects. Analysis of these cases showed that definite field loss was preceded by increased variability of measured threshold values. Already at this early stage up to the time when a definite diagnosis of field loss first could be made, was, in all eyes, the disturbed area large enough to cover several test locations in the standard test point pattern used during the study. The results indicate that test point density of standard central field automated threshold tests is entirely sufficient, and that early field loss is unlikely to be extremely focal in nature and able to escape detection between tested points. While early recognition of glaucomatous field loss obviously is facilitated by access to large amounts of threshold data, the current results imply that test point density is usually not a limiting factor, but that testing an eye twice using the same standard test point pattern might be as effective as if two interlocking grids were used. PMID- 8249573 TI - Differences of retinal nerve fiber layer thickness between normal and glaucoma like optic disks (physiological cups) matched by optic disk area. AB - We matched by disk area size 23 eyes of patients with glaucoma-like disks (physiological cups) with normal ocular pressures and no visual field loss with 23 eyes of normal subjects. Using stereophotogrammetry we found a significant decrease in retinal nerve fiber layer thickness in the eyes with glaucoma-like disks compared to the normal eyes (P = 0.0081), especially in the nasal quadrant (18.1%). A similar significant decrease was noted in the neuroretinal rim area and a significant increase in cup volume, area, depth and slope for the total disk and its quadrants. The use of computerized image analysis also showed a significant increase in pallor area for glaucoma-like disks compared to normal disks. Eyes with glaucoma-like disks or physiological cups showed evidence of an optic neuropathy and may be manifesting the same damage to the optic nerve occurring in high pressure open angle glaucoma. PMID- 8249574 TI - The effect of betaxolol and timolol on postoperative intraocular pressure. AB - Postoperative intraocular pressure (IOP) elevation is a well documented phenomenon after cataract surgery. Many pharmacological agents have been used to prevent this complication. We conducted a randomized double-blind, placebo controlled study to evaluate the effect of topical betaxolol and timolol on postoperative IOP after extracapsular cataract extraction and intraocular lens implantation. One hundred and twenty patients were randomly divided into three groups. At the end of the operation, the patients were given one drop of 0.5% betaxolol, 0.5% timolol or placebo solution into the lower cul-de-sac of the operated eye. IOP was measured preoperatively and five and twenty-four h postoperatively. There was a significant rise in IOP in the placebo and betaxolol groups but not in the timolol group five h postoperatively. IOP was significantly higher in all groups twenty-four h after the operation than preoperatively. Betaxolol did not have any significant effect on the postoperative IOP, but timolol was effective in preventing the early IOP rise. The differences in the antagonist effects of these drugs and the possible role of beta 2-receptors in mediating the aqueous humour flow are discussed. PMID- 8249575 TI - Normal tension glaucoma: the value of predictive tests. AB - Fifty-four normal tension glaucoma cases were studied to determine the value of several clinical tests for predicting the progression of the disease. Outflow facility, intraocular pressure (IOP) increase after water drinking, and diurnal changes in IOP were studied. Progression was determined on the basis of changes in visual sensitivity as measured on the Octopus 201. A minimum of four examinations of the central 30 degrees were conducted over a 3- to 7-year period. Progression of visual field defects was seen in 38.5% of eyes that had demonstrated some degree of abnormality in at least one of three clinical tests, while only 10.7% of those eyes that appeared normal on the basis of these tests showed such progression. The difference was significant (p < 0.04). These results suggest that the three clinical tests may be of value in detecting normal tension glaucoma eyes at risk for progression of visual field defects. PMID- 8249576 TI - Glare measurements before and after cataract surgery. AB - The efficacy of cataract surgery in improving visual performance was evaluated in 10 patients by measuring changes in intraocular light scatter, contrast sensitivity and glare induced visual loss. Two different methods were used to estimate these functions before and after cataract extraction with implantation of a monofocal posterior intraocular lens. By one method intraocular light scattering was assessed. By the other, letter contrast sensitivity and glare induced visual loss were determined. Preoperatively most patients had various glare problems, glare induced visual loss and increased intraocular light scatter. Following surgery the letter contrast sensitivity increased by a factor of 3 or more (mean 4.3). The glare problems diminished or disappeared after surgery, although several patients still had increased intraocular light scattering and glare induced visual loss compared to normals. Postoperatively the contrast sensitivity both with and without glare had increased to a level where the glare induced visual loss did not create any visual problems. PMID- 8249577 TI - Cataract surgery. Computer-based registration and analysis of data. AB - Consecutive registration, rapid retrieval and relevant analysis of information about cataract patients were obtained by applying a data form. The form was designed to be used with a statistical computer program (SPSS/PC). Data describing cataract cases, operations and postoperative controls were recorded. The information is contained in 76 variables, each with up to 10 alternative answers. The main advantages are the opportunity for quality control and clinical research. As an example, an analysis of the pre- and postoperative visual acuity is presented. Originally, the material consisted of 969 eyes, and of these, 948 eyes were available for follow-up. The postoperative corrected visual acuity was 5/5 or better in 57.2% of the eyes. In 80.4% of the eyes a visual acuity better than 5/10 was achieved. The main cause of postoperative visual acuity less than 5/8.5 was age-related macular degeneration (115 eyes). This maculopathy was recognized preoperatively in 76 eyes. PMID- 8249578 TI - Secondary implantation of flexible open loop anterior chamber IOLs. AB - Secondary implantation of flexible, open loop anterior chamber IOLs was performed on 20 aphakic eyes between January 1990 and May 1992. In 3 cases anterior vitrectomy was necessary. No other complications were encountered during surgery. The patients were followed up for 4 months postoperatively. Seventeen eyes achieved a corrected visual acuity of 5/5 or better. Fourteen eyes achieved the same visual acuity as preoperatively, and 5 ended with an improved visual outcome. In one case the visual acuity became worse than preoperatively due to cystoid macular edema. No other serious postoperative complications occurred. The postoperative refraction was within 1 diopter of emmetropia in 16 eyes compared to the estimated (biometry) 19 eyes. We conclude that implantation of flexible open loop anterior chamber IOLs is a safe procedure in properly selected cases. PMID- 8249579 TI - Full-field electroretinogram in a patient with cutaneous melanoma-associated retinopathy. AB - A patient operated for malignant melanoma developed night blindness and a sensation of shimmering light, strong enough to make him unable to work. Shortly afterwards, melanoma metastases became apparent. A full-field ERG examination showed absence of rod responses but normal cone amplitudes, suggesting a malfunctioning rod system in the retina. The patient was not on chemotherapy. A few similar cases have previously been reported. The disease appears to differ from the cancer-associated retinopathy (CAR) syndrome, which affects mainly the photoreceptor cells. PMID- 8249580 TI - Full-field electroretinograms during general anesthesia in normal children compared to examination with topical anesthesia. AB - Full-field electroretinograms were obtained from 20 normal children during general anesthesia and compared with electroretinograms obtained with topical anesthesia. Disoprofol and fentanyl induced a reduction in the b-wave amplitudes and an increase in the cone b-wave implicit time, but the changes were small and fall within the range limits seen with topical anesthesia (i.e., average +/- 2 SD). PMID- 8249581 TI - Simultaneous occurrence of primary choroidal and cutaneous malignant melanoma and skin metastasis. AB - This case report describes the clinical and histopathological findings in a 65 year-old woman enucleated for a malignant choroidal melanoma. Simultaneously, an excision was performed of a cutaneous melanoma together with a satellite nodule presumed to be a metastasis from the cutaneous, superficial spreading melanoma. Six months later, chemotherapy for liver metastasis was given without effect. There were no signs of dysplastic nevus syndrome. A 39-year-old cousin, however, had been enucleated for a malignant choroidal melanoma. This sporadic case might suggest a common etiologic factor in the pathogenesis of multicentric melanomas. PMID- 8249582 TI - Survey of eye injuries in Norwegian children. AB - A review of the medical records of 238 children younger than 16 years admitted with ocular injury to the University Hospital in Trondheim during a 10-years period was undertaken to provide information about the causes, circumstances and visual outcome of ocular trauma in young patients. Children with ocular injury represented 14% of all paediatric eye admissions. The majority were boys (77%). The frequency of injuries among boys increased markedly from the age of 8 years, while the frequency was almost the same among girls in all age groups. The most common cause of injury was projectiles (21.5%) followed by sticks, twigs and pencils (10.1%), falls (10.1%), bow and arrows and catapults (9.7%) and balls (8.8%). The most frequent diagnosis was contusion (43%). Perforating eye injuries amounted to 19%. Follow-up examination showed that 49% of children with eye injuries had some visual deficit, including eighteen children (8%) with visual acuity worse than 0.1. Because most of the eye injuries among children are preventable, more appropriate strategies for the prevention of these should be implemented. PMID- 8249583 TI - Three-dimensional structure of human retinal vessels studied by vascular casting. AB - A new casting technique for studying the three-dimensional structure of the human retinal vascular system on post mortem tissue was developed. The technique was designed to provide an overview presentation of the retinal vasculature such that localized areas with vascular pathology could be identified for further analysis of three-dimensional structure. The paper presents qualitative characteristics of three-dimensional structure in the normal human retinal vascular bed, to provide a basis for future studies on vascular impairment in retinal disease. PMID- 8249584 TI - Colour vision in diabetic patients after photocoagulation treatment. A five-year follow-up. AB - Colour vision of 60 diabetic patients (60 eyes) was studied after photocoagulation treatment in 1986-87. For the follow-up study 5 years later in 1991-92, 32 of the patients were available. The ages of the patients in the follow-up study varied from 28 to 64 years, the duration of diabetes from 19 to 35 years, the amount of laser spots from 200 to 3174, and the visual acuity from 0.4 to 1.0. As colour vision tests, the Standard Pseudo-isochromatic Plates part 2, Lanthony Tritan Album, Farnsworth Panel D 15, and box III of the Farnsworth Munsell 100 hue test were used. Of the 32 eyes, 22 (68.8%) had the same results in the colour vision tests, 4 (12.5%) had better results, and 6 (18.8%) had worse results than 5 years earlier. Between Group 1 (colour vision the same or better than 5 years earlier, 26 eyes) and Group 2 (colour vision worse than earlier, 6 eyes) there was a significant difference in the age and in the level of the visual acuity of the patients. The changes in the lens, fundus or visual acuity during 5 years did not seem to have an effect in colour vision. PMID- 8249585 TI - Low-grade myopia in children with regressed retinopathy of prematurity. AB - Myopia is a frequent sequelae of retinopathy of prematurity. The mechanism for myopia development in children born preterm is not well understood. From a group of 33 myopic preterm children we selected those who had regressed retinopathy and myopic values < or = 5 D. Seventeen children met our criteria. Of them, 10 patients could be re-examined. Factors related to myopia development were analyzed and compared to a control group of 12 children born fullterm with emmetropia or low hypermetropia, as well as another group of 10 fullterm children with myopic values < or = 5 D. The age of children ranged from 9 to 14 years in all groups. The mean birth weight in myopic premature children was 1249.3 g. Higher keratometric diopter values were found in myopic premature children than in emmetropic fullterm children (p < 0.05) and myopic fullterm children (p < 0.05). Increased axial length values were observed in myopic fullterm children when compared to myopic premature children (p < 0.05) In children born preterm the corneal refractive power seems to play a major role in myopia development. PMID- 8249586 TI - Human basic tear fluid osmolality. I. Importance of sample collection strategy. AB - Osmolalities of 200 human tear prism fluid samples collected from two subjects were determined from their melting-point temperatures with the Clifton Nanoliter Osmometer by calibration with 200 standard solution samples (290 mOsm/kg). Comparisons were made between tear fluid osmolalities obtained using a single sample simultaneous-recalibration method: 1) for tear samples collected using finely-drawn microcapillaries without biomicroscopy, vs secondly with biomicroscopic observation by illumination of only the sampling area on the inferior tear prism, the two collections separated by an interval of 10 min; and 2) for samples collected without biomicroscopy before, vs after a 10-min interval. Tear fluid collection using a biomicroscope resulted in values that were significantly lower than those collected without (overall mean = 299.5 and 306.6 mOsm/kg, respectively; p < 0.0001). The difference (7.1 mOsm/kg) may have resulted from mechanical, photic, and/or psychogenic reflex stimulation due to biomicroscopy, as the 10-min interval had no osmotic effect on samples collected without biomicroscopy. Based on these results from two subjects, we suggest that relatively rapid, repetitive collections of human tear prism fluid can be made without significantly disturbing the osmotic outcome. However, physiological representation of basic human tear fluid is more accurate by avoidance of reflex inducing collection methods that were formerly considered unobtrusive. Basic human tear prism fluid is more hypertonic, by at least 7 mOsm/kg, than generally realized. PMID- 8249587 TI - Human basic tear fluid osmolality. II. Importance of processing strategy. AB - Osmolalities of 100 human inferior tear prism fluid samples collected from two subjects and 100 standard solution samples (290 mOsm/Kg) were determined from their melting-point temperatures with the Clifton Nanoliter Osmometer. Accuracy and reliability comparisons were made for endpoints obtained using a single sample simultaneous-recalibration (SS/SR) strategy, vs a lowest-of-six periodic recalibration (LS/PR) strategy. Tear fluid osmolality values based on the LS/PR strategy were significantly lower (hypotonic) than for the SS/SR strategy (overall mean = 302.4 and 307.8 mOsm/Kg, respectively; p < 0.0012). The mean difference (5.4 mOsm/Kg) resulted from the combined effects of machine drift and reduced reliability of endpoint determination for tear fluid in comparison to that of a homogenous standard solution. LS/PR osmolality was linearly correlated to SS/SR osmolality with a slope of 0.433 and the two endpoint strategies resulted in equivalent values at 298.4 mOsm/Kg. Eighty-four percent (84%) of individual osmolality readings were greater than 298.4 mOsm/kg, and an 'averaging type' effect caused LS/PR osmolalities to be increasingly less than SS/SR values above this value. These outcomes from two subjects were approximated by a parallel statistical model. We suggest that physiological representation of basic human tear fluid is more accurate using the single-sample simultaneous recalibration strategy. Basic human tear prism fluid is more hypertonic and has a greater within-subject range in normal (non-dry eye) humans than previously realized. PMID- 8249588 TI - Topographical corneal edema. An update. AB - Measurement of the topographical corneal edema response has been previously restricted by the use of optical pachometry and small sample sizes. This study uses ultrasound pachometry to measure changes in the corneal thickness of 23 subjects at 9 locations during 2 hours' wear of a thick soft contact lens. The data are analysed using both averaged and individual location data. The averaged location analysis supports the current paradigm that less edema occurs in the peripheral cornea than in the central cornea during contact lens wear. However, the individual location analysis reveals a more complex corneal edema response in which the greatest amount of edema following 2 hours' contact lens wear occurred in the central and mid-peripheral temporal cornea, in comparison with the peripheral nasal and peripheral inferior cornea which showed the least edema. The edema measured in the peripheral temporal and peripheral superior cornea following 2 hours' contact lens wear was not significantly different from that measured in the central cornea. The edema measured in the central cornea following 2 hours' contact lens wear was not significantly different from that found at any mid-peripheral location. This study demonstrates that the corneal edema response associated with 2 hours' wear of a thick soft contact lens is a complex dynamic topographical process and cannot be simply modelled using averaged data from small samples. PMID- 8249589 TI - Effects of retrobulbar bupivacaine on post-operative pain and nausea in retinal detachment surgery. AB - A prospective double-masked clinical trial was conducted to determine whether retrobulbar bupivacaine block had an effect on post-operative pain, nausea and intra- and post-operative use of analgesics in retinal detachment surgery performed under general anaesthesia. Thirty-two patients were randomized to have general anaesthesia with or without retrobulbar bupivacaine. Pain score was documented as 0-10 (0 = no pain. 10 = worst pain ever felt). Post-operative pain score was significantly lower in the retrobulbar block group during the first post-operative hours than in the control group. Men complained more about post operative pain than did women. The patients in the retrobulbar group complained less about nausea. Significantly fewer patients in the retrobulbar group required parenteral pain relief during operation and the first 48 h after. PMID- 8249590 TI - Immunoglobulins in granular corneal dystrophy Groenouw type I. AB - Three patients with granular corneal dystrophy Groenouw type I underwent corneal grafting, and cryostat sections of the corneal buttons were examined immunohistochemically for immunoglobulins. Positive results were obtained for IgG, Kappa-, and Lambda chains with immunofluorescence technique. The reactions were seen exclusively in the same localizations as the Masson trichrome positive deposits. PMID- 8249591 TI - Differential diagnosis between granular corneal dystrophy Groenouw type I and paraproteinemic crystalline keratopathy. AB - A case of corneal opacities in a leukemic patient with an M-component in the serum proteins is presented, and a comparison is made to patients with granular corneal dystrophy Groenouw type I. The corneal deposits associated with the two conditions may appear identical with slit-lamp biomicroscopy. Granular dystrophy patients, however, show a normal serum immunoglobulin pattern in contrast to patients with paraproteinemic crystalline keratopathy. The two entities can therefore be distinguished from each other by a serum electrophoresis. PMID- 8249592 TI - Hypertonic saline test for ophthalmic nerve impairment. AB - In order to compare the accuracy of methods for testing ocular surface sensitivity (e.g. function of the first branch of the trigeminal nerve) three different methods were compared in patients with unilateral acoustic neurinomas. The three methods 1) hypertonic (3%) saline in the conjunctival sac (apparently not previously described in the literature) 2) esthesiometer (Cochet & Bonnet) touching of the cornea, and 3) touching the cornea with a cotton wool wisp, was found to reveal reduced ocular surface sensitivity on the neurinoma side in 50% (hypertonic saline), 23% (esthesiometer) and 14% (cotton wool wisp) of cases, respectively. With McNemar's test for comparing test methods the 3% saline test proved significantly more sensitive than the cotton wool wisp test (p < 0.05), but not significantly more sensitive than the esthesiometer test (p > 0.10). The advantage of the 3% saline test, apart from its high sensitivity, is that it does not require sterilization of any equipment as is the case for the esthesiometer nor, in contrast to the methods using corneal or conjunctival touch, does it require perfect visual control by means of a magnifying glass in order to be performed accurately, and it is not affected by visual stimuli. It should therefore be the preferred test of assymetry in ophthalmic nerve function. PMID- 8249593 TI - Healon GV in posterior segment surgery. AB - Healon, a viscoelastic preparation of Sodium hyaluronate, has been reported effective in posterior segment surgery for dissection of epiretinal membranes, i.e. proliferative diabetic retinopathy and macular pucker. A more viscous substance might be more effective for this kind of surgery. Therefore, a new Sodium hyaluronate preparation Healon GV with a ten times greater viscosity at rest than Healon, was tested in an open clinical study. Twenty-four patients of both sexes and not younger than 18 years were enrolled. The follow-up time was three months. The main indications for using Healon GV were: to separate membranes from the retina, to stop bleeding and to reposition a detached retina. 0.2-0.5 ml of Healon GV was injected through a 27 gauge bent cannula and at close of surgery irrigated out of the eye. Postoperatively the patients were monitored concerning intraocular pressure, vitreous clarity, anterior chamber flare and visual acuity. Healon GV was found to make the surgical procedure simpler and safer. No inflammatory reaction and no increase of IOP was observed. PMID- 8249595 TI - How to judge the efficacy of glaucoma treatment. PMID- 8249594 TI - Multiple melanocytic tumours in a case of ocular melanocytosis. AB - A case of ocular melanocytosis with the simultaneous occurrence of a choroidal malignant melanoma, a ciliary body melanocytoma and a surface plaque iris naevus in an 80-year-old male Caucasian is reported. Patients with ocular or oculodermal melanocytosis are considered to be at risk for tumour formation and malignant transformation; the multiple melanocytic tumours of the present case support this hypothesis. PMID- 8249596 TI - A rare corneal injury caused by molten lead. PMID- 8249598 TI - [Direct measurement of oxy-radicals using HPLC-ECD]. AB - A direct, sensitive, simple and specific high-pressure liquid chromatographic (HPLC) method was used for the quantitation of hydroxyl radicals by means of oxy radical trapping of DMPO to form DMPO-OH adducts. The DMPO-OH adduct peak was separated successfully and identified by HPLC-ECD with a Waters ODS reversed phase 10 microns column. The mobile phase composed of citric acid/sodium acetate (citric acid-30 mmol/L-sodium acetate 50 mmol/L-3% acetonitrile, pH 5.1), at a flow rate of 1.2 ml/min and detection potential of 0.6 V with Ag/AgCl as reference electrode. Both EDTA-Fe(2+)-H2O2 (FeSO4 300 mumol/L, EDTA 300 mumol/L, H2O2 180 mumol/L and DMPO 2 mmol/L) and H2O2 photolysis (H2O2 18 mmol/L and DMPO 2 mmol/L photolysis for 6 min) systems were taken to produce hydroxyl free radicals for screening new drugs and studying the mechanism of action. The relative standard deviations were 6.1 and 8.0% respectively. The sensitivity of the method was shown to be similar to that of ESR. The method for detection of superoxide anions with HPLC-ECD was also described. PMID- 8249597 TI - [Estimation of free calcium level within synaptosomes by using fura-2 and the effect of calcium channel agonist and antagonist]. AB - The cytosolic free calcium concentration [Ca2+]i in synaptosomes was determined with the fluorescent indicator fura-2, the effects of calcium channel agonist and antagonist on intracellular Ca2+ level were studied. The cytosolic ionized calcium concentrations in resting status were between 200 nmol/L and 400 nmol/L. Cytosolic Ca2+ was elevated following increases in Ca2+ concentration in the medium, plasma membrane depolarizations induced by KCl, and the addition of glutamate and NMDA. On the other hand, the increase of cytosolic Ca2+ induced by KCl was decreased by verapamil and that induced by NMDA was decreased by MgCl2. A few critical problems in [Ca2+]i detection were also discussed. PMID- 8249599 TI - [Synthesis and antitumor activities of 4-acylamido-4-deoxy-4' demethylepipodophyllotoxin analogues]. AB - Twenty four 4-acylamidc-4-deoxy-4'-demethylepipodophyllotoxins were synthesized by acylation of 4-amino-4-deoxy-4'-demethylepipodophyllotoxin by means of usual methods, and screened in vitro. This type of compounds showed strong antitumor activities, most of them are superior to etoposide in the inhibition against L1210 and KB cells. PMID- 8249600 TI - [Studies on total synthesis of squamosamide]. AB - A new natural compound squamosamide isolated from Annona squamosa, was synthesized. This compound was characterized by 1H, 13CNMR and MS. PMID- 8249601 TI - [Isolation and structure elucidation of qianhucoumarin A]. AB - Seven compounds were isolated from the traditional Chinese Medicine Qianhu, roots of Peucedanum praeruptorum Dunn. Their structures were identified as psoralen (II), 5-methoxypsoralen (III), 8-methoxypsoralen (IV), pteryxin (V), peucedanocoumarin II (VI), beta-sitosterol (VII) and qianhucoumarin A (VIII) by physico-chemical constants and spectral analyses (UV, IR, 1HNMR, 13CNMR, EIMS). VIII is a new compound with the structure of 3'(R)-hydroxy-4'(R)-tigloyloxy-3',4' dihydroseslin. Its absolute configurations were determined by chemical correlation with known compounds. II, III, IV and VII were first isolated from this plant. PMID- 8249602 TI - [Studies on the chemical constituents of Populus tomentosa carr]. AB - Populus tomentosa Carr. a Chinese folk medicine, has been used for treatment of arthritic disease. A new flavonoid(1), with molecular formula C30H26O12 was isolated from this plant along with four known compounds. They were identified as apigenin-7-O-(6"-O-p-coumaroyl)-beta-D-glucopyranoside (I), salicin (II), tremulodin (III), tremulacin (IV) and daucosterol (V) by the use of 1HNMR, 13CNMR and 1H-13C COSY spectroscopy. PMID- 8249603 TI - [The crystal structure atlas data base system for chemical constituents of Chinese traditional and folk medicine]. AB - The atlas data base system for chemical constituents of Chinese traditional and folk medicine is the first one for single crystal structure in China. It includes about 250 crystal structures of chemical ingredients of Chinese traditional and folk medicine. It has searching, plotting and computing functions. It is a useful reference base. All softwares are written in dbaseIII, FORTRAN and Assembler Languages. They can be run on PC-286,386,486 and their compatible microcomputers. PMID- 8249604 TI - [Determination of the optical purity of praziquanamine by reversed phase HPLC after derivatization with GITC]. AB - The enantiomers of praziquanamine were separated by reversed phase HPLC after derivatization with 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosylisothiocyanate (GITC). The determination of the optical purity of praziquanamine was successfully achieved. This method was applied to the determination of the optical purity of d- and l-praziquanamines. PMID- 8249605 TI - [Studies on distribution of magnetic gelatin microspheres in rabbits]. AB - In this report, the technique of labelling MG-ms with 99mTc as pertechnetate in the presence of a reducing agent such as SnCl2 was described. The distribution of intravenously injected 99mTc-labelled MG-ms in rabbits at different intervals of magnetic field applied and different magnetic field intensity was investigated by using an externally applied magnetic field and measuring the radioactivity at the rabbit head and other organs. When magnet was used, the radioactivity in the head, target site, was 15 times more than that when magnet was not used. At the same time, the radioactivity of the lung was 5 times less than when magnet was not used. The newly designed magnetic field equipment was presented. PMID- 8249606 TI - [Effect of dihydroepiandrosterone sulfate on the tension of uterine cervix and its mechanisms]. AB - After i.v. injection of dihydroepiandrosterone sulfate (DHA-S) at 20 or 40 mg/kg to female rats on day 19 of gestation, the tension and hydroxyproline level of uterine cervix were decreased obviously. However, DHA-S at 10 mg/kg showed no effect on the tension of uterine cervix. DHA-S in vitro, at 0.1 mg/kg significantly increased estradiol secretion in the ovaries and placenta of late pregnant rats. PMID- 8249607 TI - [Effect of seleno-organic compounds on leukotriene B4 biosynthesis]. AB - Leukotrienes (LTs) are a group of metabolites of arachidonic acid through the 5 lipoxygenase pathway. Among these metabolites, LTB4 is an important mediator of inflammatory disease. Recently, it has been shown that seleno-organic compounds are very biologically active. One of them, Ebselen [2-phenyl-1,2 benzoisoselenazol-3 (2H) one] is a new seleno-organic compound with very low toxicity while exhibits anti-inflammatory activity. Attempt to search for seleno organic compounds as anti-inflammatory drugs and establish structure-activity relationships, ten ebselen derivatives with modifications in the 2-phenyl moiety were studied with respect to their effects on LTB4 biosynthesis. p-substituted compounds were shown to have stronger inhibitory activity on LTB4 biosynthesis than o-substituted compounds and ebselen itself. Among the p-substituted compounds, polar-inducing group-substituted compounds showed stronger activity than compounds substituted with polar-conjugated groups. Among the compounds substituted with polar-inducing groups, strongpolar groups exhibited stronger activity than weakpolar groups. PMID- 8249608 TI - [Synthesis and antibacterial action of 1-ethyl-6-fluoro-1,4-dihydro-4-oxo-7-(4 aroylthiocarbamoyl-1-piperaziny l)-3-quinoline carboxylic acids]. AB - Thirteen new 1-ethyl-6-fluoro-1,4-dihydro-4-oxo-7-(4-aroyl-thiocarbamoyl-1 piperazinyl)-3- quinoline carboxylic acids were prepared. Their structures were characterized by elemental analysis, IR, HNMR and MS spectra. Preliminary pharmacological tests indicated that some of compounds Ia-m possess strong inhibiting activity against Escherichia coli, Bacillus subtilis and Proteus at concentration of 100 micrograms/ml. PMID- 8249609 TI - [Main carotenoids in pressed seeds (Cucurbitae semen) of oil pumpkin (Cucurbita pepo convar. pepo var. styriaca)]. AB - Various use of the oil-pumpkin offers reason for the phytochemical analysis of seed-meal's carotene pigments. Column chromatography was performed on the adsorbents MgO, Celite and CaCO3 with hexane and benzene as eluents. HPLC separation of different pigments was carried out on a 6 microns reverse phase packing with a ternary gradient elution method using a diode-array detector. The main components of the press-residue were lutein [3,3'-dihydroxy-alpha-carotene = (3R,3'R,6'R)-beta,epsilon-carotene-3,3'-diol; 52.5%] and beta-carotene (beta,epsilon-carotene; 10.1%). In addition to the above-mentioned pigments it was successful to reveal the presence of violaxanthin, luteoxanthin, auroxanthin epimers, lutein epoxide, flavoxanthin, chrysanthemaxanthin, 9(9')-cis-lutein, 13(13')-cis-lutein, 15-cis-lutein (central-cis)-lutein, alpha-cryptoxanthin, beta cryptoxanthin and alpha-carotene (beta,epsilon-carotene) in small quantities. PMID- 8249610 TI - Dissociation constant of chlordiazepoxide in montmorillonite suspension and its pharmaceutical application for a controlled-release dosage form. AB - The changes in the distribution of the ionic species of a drug due to interactions with a clay surface may have important consequences in drug therapy. The binding of chlordiazepoxide (CDO) into montmorillonite clay, veegum: HV (V), surface was studied under several pH conditions. In vitro drug release behaviour from CDO-V products was investigated. Changes in the partial molar free energy of the ionic species of CDO, delta Gi (i = CDO, CDOH+) as a result of interaction with V were determined by using differential absorbance spectroscopic techniques at 260 and 310 nm. The stronger interaction between the negatively charged clay surface and the protonated species (CDOH+) relative to the parent compound is responsible for the apparent displacement of its pKa value in V suspensions from 4.64 to 5.67. In vitro drug release behaviour from CDO-V product to the simulated gastrointestinal fluids showed a controlled pattern. The in vivo results indicated a prolonged drug effect after the oral administration of drug-clay preparation. PMID- 8249611 TI - [Formulation of diazepam suppositories, results of rheological and biopharmaceutical studies. 1. Rheological study of the used suppository bases]. AB - The rheological properties of four lipophilic and three hydrophilic suppository bases were studied. The flow and viscosity curves, the initial, equilibrium and plastic viscosity as well as the Bingham's yield value of the systems were determined. It was found that the viscosity curves had two different parts in the examined shearing rate range: the part of greater rise described a definitive destruction of the system, while the part of smaller rise represented an equilibrium state. The values of the rheological parameters of the lipophilic and hydrophilic systems showed a definite difference, which was explained by the greater association of the hydrophilic bases. PMID- 8249612 TI - [Formulation of diazepam suppositories, results of rheological and biopharmaceutical studies. 2. In vitro membrane diffusion and in vivo absorption results]. AB - In the first part of the publication the rheological properties of the vehicles used for the production of suppositories were studied in order to determine the ideal parameters for formulation. In this part a detailed methodology and the results of the in vitro membrane diffusion and in vivo bioavailability studies, are presented. The results confirm a general correlation between the in vitro and the in vivo findings. It seems that hydrophilic macrogol-mixture with great molecular mass can be recommended as the optimal vehicle for formulation of diazepam suppositories. PMID- 8249613 TI - Conditions of cyclodextrin complexation. AB - In the past 20 years, cyclodextrin (CD) research has achieved considerable results, as indicated by the large number of publications and patents, the six international symposia on CDs, the new dosage forms of medicines prepared with CDs, etc. Sufficient data have accumulated to permit a survey of the most important conditions of CD complexation. Further, an account is presented of the success of precipitation of the complex involving furosemide and beta-CD. PMID- 8249614 TI - [Comparative evaluation of dissolution and membrane transport methods of suppositories]. AB - The dissolution and transport rate constants of Diaphyllin, Feldene, Naprosyn and Tilcotil suppositories originated from the same batches were studied. The dissolution tests were carried out with Desaga type flow cell. The membrane transport experiments were performed in "Muhlemann-tester" and in the own designed "membrane-diffusion cell". The values of the dissolution rate constants were higher than the diffusion rate constants. The results of the same batches of the suppositories tested in Muhlemann-tester or in the own designed membrane diffusion cell were in good correlation. On the basis of the experiments carried out by the membrane-diffusion cell we can state that it is able to be standardised, automated and the results show good reproducibility. PMID- 8249615 TI - Combined action of phenobarbital and dexamethasone on the activity of rat liver P450 system. AB - Phenobarbital and dexamethasone are potent inducers of the same cytochrome P450 form, CYP3A1, but the mechanism of action is not quite clear. If the mechanism of induction by phenobarbital and dexamethasone is different, additive effect may be observed in the specific activities of CYP3A1: ethylmorphine or aminopyrine N demethylation of liver microsomes from rats treated with phenobarbital and dexamethasone in combination. The results of recent work could not display differences in CYP3A1 activities between the groups of animals single administered and coadministered with phenobarbital and dexamethasone. However, p nitrophenol hydroxylation surprisingly increased threefold as a result of double induction. 3-amino-1,2,4-triazole inhibition study shows that CYP2E1 is responsible for 60% of enhanced p-nitrophenol hydroxylase activity, but the residual 40% indicates the participation of other isoenzyme(s). PMID- 8249616 TI - Possible involvement of nitric oxide in arginine-induced analgesia. AB - Intracerebroventricular injection (icv) of 1-arginine (Arg, 0.5, 5.0, 50 micrograms) produced a dose-dependent prolongation in the hot-plate latency in mice. A similar result was obtained when nitroprusside (0.1, 1.0, 10 micrograms, icv) was given. Naloxone (2 mg.kg-1, ip) failed to antagonize the effects of Arg or nitroprusside. But Arg-induced antinociception was attenuated by NG-monomethyl arginine (NMMA, 10 micrograms). 8-Bromoguanosine 3': 5' cyclic monophosphate (bromo-cGMP, 50 micrograms) also produced an antinociceptive effect. The results suggest that Arg induces an analgesia possibly via activation of nitric oxide cGMP pathway. PMID- 8249617 TI - Specific binding of [125I]iodomelatonin in pigeon and quail spleen membrane preparations and effect with hydrocortisone-treatment. AB - [125I]Iodomelatonin binding sites were studied in the spleens of pigeon and quail. Scatchard analyses of the membrane preparations collected at midday (12:00) revealed an equilibrium dissociation constant (Kd) of 250 +/- 74 pmol.L-1 and a maximal number of binding sites (Bmax) of 13.4 +/- 1.8 fmol/mg protein in the pigeon spleen and Kd = 390 +/- 132 pmol.L-1, Bmax = 21.3 +/- 3.7 fmol/mg protein in the quail spleen. Circadian studies indicated that spleens collected at midnight (24:00) had 53% and 70.9% less [125I]iodomelatonin binding sites than the samples collected at midday in the pigeon and quail, respectively. Specificity studies showed only melatonin and 6-chloromelatonin had significant inhibition to [125I]iodomelatonin binding. Hydrocortisone 15 mg.kg-1 x d-1 im for 5 d selectively increased [125I] iodomelatonin binding sites in pigeon spleen. These results suggest that spleen is a site of melatonin action outside the brain. PMID- 8249618 TI - Pharmacokinetics and relative bioavailability of nimodipine capsules and tablets in 8 Chinese healthy men. AB - A single oral dose of tablets or capsules of 239 mumol nimodipine was given to 8 healthy volunteers of Han nationality in a randomized crossover study. Plasma levels were determined with HPLC method. The plasma concentration-time curve fitted to a first order absorption, 1-compartment open model, and the T1/2K was around 2 h. Although the capsules could reach peak level faster, the bioavailability was not significantly different from that of the tablets. PMID- 8249619 TI - Determination of beta-adrenoceptor subtypes in human pulmonary artery and thoracic aorta by radioligand binding. AB - The beta-adrenoceptors of human pulmonary artery (PA) and thoracic aorta (TA) were studied by the use of a high specific activity radioligand [125I]pindolol (Pin). To identify the subtypes of beta-adrenoceptors in the 2 blood vessels, the competitive inhibition curves of [125I]Pin by beta 1-antagonist atenolol and beta 2-agonist salbutamol were analyzed using a computer program LIGAND of a mathematical model of the ligand-binding system. The Bmax (15.3 +/- 1.2 fmol/mg protein) and the dissociation constant Kd (44 +/- 4 pmol.L-1) for PA were similar to those for TA (Bmax, 12.8 +/- 1.2 fmol/mg protein, Kd, 45 +/- 4 pmol.L-1). Competitive inhibition analysis showed that the beta 1- and beta 2-adrenoceptor subtypes coexisted in human PA and TA, with beta 1 adrenoceptor dominant. The ratios of beta 1:beta 2 in PA and TA were 2.9:1.0 and 2.1:1.0, respectively. PMID- 8249620 TI - Antifibrillatory effect of tetrahydroberberine. AB - Electric stimulation and drug-induced ventricular fibrillation (VF), monophasic action potentials (MAP), and triggered activity were studied before and after administration of tetrahydroberberine (THB) in rabbits, rats or guinea pigs. At doses of 5, 10, and 20 mg.kg-1, i.v. THB increased the ventricular fibrillation threshold, and the BaCl2-induced VF was also prevented or terminated by THB in rabbits. Centrogenic VF induced by icv aconitine in rats was inhibited by pretreatment with THB in a dose-dependent manner, whereas VF induced by iv ouabain in guinea pig was inhibited to a lesser degree. For MAP, the duration at 90% repolarization (MAPD90) was prolonged remarkably, whereas the MAPD20, the MAP amplitude, and the maximal velocity of phase 0 were shortened or decreased slightly. The amplitudes of early afterdepolarization produced by cesium chloride (CsCl) were attenuated, while the cumulative threshold doses of CsCl for sustained ventricular tachycardia were elevated by THB. These results indicated that THB had an potent antifibrillatory effect, which might be attributed to its blockade of potassium, calcium, and sodium currents. PMID- 8249621 TI - Central expression of c-fos protein after peripheral noxious thermal stimulation in awake rats. AB - This study applied immunohistochemistry method to examine the pattern of c-fos expression in the neuraxis following peripheral noxious thermal stimulation accomplished by immersion of tail of awake rat into hot water (50 degrees C). In unstimulated control rats, no obvious baseline expression of c-fos protein was found except in nucleus paraventricularis hypothalami and colliculus inferior, probably associated with restraint-induced stress and auditory stimulus, respectively. Noxious thermal stimulation resulted in the activation of c-fos expression, and bilateral increased nuclear immunostaining was counted in dorsal horn of lumbar and sacral segments of spinal cord (laminae I, II), nucleus raphe dorsalis, substantia grisea centralis (ventralis), nucleus paraventricularis thalami, nucleus anterior thalami, nucleus ventralis thalami, nucleus medialis thalami, nucleus reuniens, nucleus rhomboideus, nucleus habenulae lateralis, nucleus paraventricularis hypothalami, nucleus arcuatus, nucleus lateralis hypothalami, nucleus preopticus lateralis, nucleus septi lateralis, nucleus amygdala, nucleus striae terminalis, nucleus tractus diagonalis, and cortex cerebri. The results demonstrated that peripheral noxious stimulation induced central c-fos protein expression in a pattern of labeling nociresponsive cells. PMID- 8249622 TI - Effects of 3,6-dimethamidodibenzopyriodonium citrate on slow inward calcium current in isolated guinea pig ventricular cells. AB - The effects of 3,6-dimethamidodibenzopyriodonium citrate (I-65) on action potentials and slow inward calcium current (I(si)) were examined on isolated guinea pig ventricular myocardial cells. I-65 (30-100 mumol.L-1) depressed the action potential duration at 20% repolarization (APD20) and, under voltage-clamp conditions, reduced the amplitude of I(si) without changing the I-V relations. I 65 also showed use-dependent effects on I(si). These suggest that I-65 may block I(si) by acting on the inactivated state of Ca channels. PMID- 8249623 TI - Effect of Panax notoginseng saponins on increased proliferation of cultured aortic smooth muscle cells stimulated by hypercholesterolemic serum. AB - Panax notoginseng saponins (PNS) was extracted from a Chinese herb medicine. After preparation of cultured aortic smooth muscle cell (SMC) from primary aortic explants, the cytotoxicity of hypercholesterolemic serum (HCS) for cultured cells was determined by trypan blue exclusion test, and [3H]thymidine incorporation and cell numbers were counted at the same time. The results showed that HCS (0.5 mg cholesterol.ml-1) increased the incorporation of [3H]thymidine into cultured cells (3722 +/- 440 vs 1655 +/- 288 dpm/microgram cell proteins, P < 0.01), stimulated the proliferation of SMC [(6.5 +/- 1.5) x 10(5) vs (4.3 +/- 1.2) x 10(5) cells/plate, P < 0.01], and that high concentration HCS (final cholesterol concentration 2 mg.ml-1) was cytotoxic to the cultured cells. PNS (100 and 400 micrograms.ml-1) decreased the incorporation of [3H]thymidine into SMC in culture with or without HCS (1292 +/- 260 and 982 +/- 314 or 4111 +/- 886 and 2361 +/- 751 dpm/microgram cell protein), and inhibited the proliferation of the cultured cells [(3.3 +/- 0.7) x 10(5) and (2.9 +/- 0.7) x 10(5) or (4.7 +/- 1.4) x 10(5) and (4.1 +/- 1.2) x 10(5) cells/plate). We conclude that PNS can inhibit the proliferation of aortic SMC stimulated by HCS. These results also suggest that HCS may play an atherogenic role in the arterial wall and that PNS may prevent atherosclerosis and inhibit progression of the atherosclerotic lesions by interfering with the proliferation of arterial SMC. PMID- 8249624 TI - Alpha 1A- and alpha 1B-adrenoceptor-mediated positive chronotropic effects on isolated rat atrium. AB - The positive chronotropic effect of alpha 1-adrenoceptor subtypes was studied on the isolated right atrium of adult male Wistar rats. In the presence of beta blocker propranolol, chloroethylclonidine (Chl) (an irreversible alpha 1B inactivator) reduced the positive chronotropic effect of phenylephrine (Phe). When the basal rate was lowered by cholinergic stimulation. Chl showed less effect on the increase of spontaneous beat rate. H-7, a protein kinase C (PKC) inhibitor, lowered the chronotropic effect to Phe under physiological condition, but did not affect the Phe-induced positive chronotropic effect when basal rate was lessened by carbachol. These suggested that the chronotropic effect was mainly mediated by alpha 1B receptors, and partially related to the activation of PKC by second messengers under physiological conditions. The compensatory effect of the alpha 1-adrenoceptors was mainly mediated by alpha 1A receptors, which is scarcely involved in the activation of PKC. PMID- 8249625 TI - Increased heart microsomal Na(+) K(+)-transporting ATPase activity by tetrandrine in spontaneously hypertensive rats. AB - In spontaneously hypertensive rats (SHR), the heart microsomal Na+, K(+)-ATPase showed a reduced activity with a higher Ca2+ sensitivity and lower responsiveness to K+. After SHR were treated with tetrandrine (Tet, ig 30 mg.kg-1, qd x 30 d) or when the microsomes obtained from SHR hearts were incubated with various concentrations of Tet, the myocardial Na+,K(+)-ATPase showed a higher activity with a decreased Km for K+, or ATP and an increased Vmax for K+. The in vitro stimulatory effect of Tet on the enzyme was dependent on the Na+ concentration in medium and was not abolished by CaCl2 1 mmol.L-1 or ouabain 0.1 mmol.L-1. However, when given in vivo, Tet tended to reduce the tolerance of the enzyme to Ca2+ and failed to change the responsiveness of the enzyme to ouabain. In vitro, Tet had similar but weaker stimulatory effect on the Na+,K(+)-ATPase from Wistar Kyoto rats. PMID- 8249626 TI - Effects of tetrandrine on production of leukotriene B4 and thromboxane B2 in rabbit blood. AB - The effects of tetrandrine (Tet) on the production of 2 major metabolites of arachidonic acid (AA), the leukotriene B4 (LTB4) and the thromboxane B2 (TXB2) in rabbit whole blood were investigated by reversed phase high pressure liquid chromatography (HPLC) and radioimmunoassay (RIA), respectively. After incubation with different doses of Tet for 15 min in vitro, the production of LTB4 and TXB2 by rabbit whole blood stimulated with calcimycin (20 mumol.L-1) was inhibited in a dose-dependent manner, with IC50 value of 17.8 +/- 8.6 and 17.7 +/- 9.2 mumol.L 1, respectively. In the presence of exogenous AA, the inhibitory effects of Tet were markedly lessened. The effects of Tet were much like those of calmodulin (CaM) antagonist fluphenazine (Flu). Dexamethasone (Dex) also inhibited the production of LTB4 and TXB2 when incubated with rabbit whole blood for 60 min. Tet iv 10 mg.kg-1 also inhibited the production of LTB4 and TXB2 in rabbit whole blood stimulated with calcimycin. These results suggest that Tet may be an antagonist of CaM, thus suppressing the release of AA which was catalyzed by CaM dependent phospholipase A2 (PLA2) from the membrane phospholipids of blood cells. PMID- 8249627 TI - Dauricine and anisodamine inhibited leukotrienes- and platelet activating factor induced DNA synthesis and proliferation of bovine cerebral microvascular smooth muscle cells in culture. AB - The effects of leukotrienes (LT) and platelet activating factor (PAF) on DNA synthesis and proliferation of bovine cerebral microvascular smooth muscle cells (BCMSMC) were studied. At 100 pmol.L-1, LTB4, LTC4, LTD4, and PAF promoted the DNA synthesis by 44%, 50%, 48%, and 57%, and enhanced the cell proliferation by 33%, 47%, 27%, and 40%, respectively. Dauricine and anisodamine inhibited the DNA synthesis of the cells induced by LT and PAF (0.1-100 mumol.L-1). These results indicate the bright future of the 2 drugs in the prevention and treatment of cerebral vascular diseases. PMID- 8249628 TI - Effects of Achyranthes bidentata polysaccharides on interleukin-1 and tumor necrosis factor-alpha production from mouse peritoneal macrophages. AB - Achyranthes bidentata polysaccharides (ABP), extracted from the root of Achyranthes bidentata, induced interleukin-1 (IL-1) synthesis as well as tumor necrosis factor-alpha (TNF-alpha) synthesis and secretion from thioglycolate primed mouse peritoneal macrophages in vitro. ABP 100-800 micrograms.ml-1 enhanced both synthesis and release of IL-1 when stimulated by lipopolysaccharides (LPS) (5 micrograms.ml-1), but had no significant influences on synthesis and release of TNF-alpha induced by LPS (10 micrograms.ml-1). Studies on IL-1 and TNF-alpha production induced by ABP (200 micrograms.ml-1) alone or plus LPS showed that peak levels of IL-1 release reached at 24 h and that of TNF-alpha release at about 2-6 h after incubation. Peritoneal macrophages from mice ip ABP 25 and 50 mg.kg-1.d-1 x 5 d produced much more IL-1 than those from control group. Peritoneal macrophages from ip ABP 100 mg.kg-1.d-1 x 6 d alone released more TNF-alpha vs control group, and showed a synergetic action with LPS (10 micrograms.ml-1), which was as strong as the positive control agent BCG. These results provide an explanation for the immunopotentiating effect of ABP. PMID- 8249629 TI - Effects of calcitriol and its analogue calcipotriol on proliferation and differentiation of human osteosarcoma cells. AB - The effects of steroid hormone calcitriol (Cal) and its analogue calcipotriol on human osteosarcoma cell line HOS-8603 were determined. When cells grew in monolayer culture in the presence of hormones, their proliferations were inhibited both in dose- and time-dependent manners. The cells showed marked morphologic changes after a 4-d treatment to apparently less transformed fibroblast-like ones. Anchorage-independent growth studies indicated that both Cal and calcipotriol at 10 nmol.L-1 inhibited colony formation by HOS-8603 cells. As a marker enzyme of the osteoblastic phenotype, alkaline phosphatase activity was induced in response to Cal or calcipotriol 100 nmol.L-1. These results suggested that Cal and calcipotriol play an important role in regulating growth and differentiation of HOS-8603 cells. PMID- 8249630 TI - Inhibitory effect of dioxopiperazine compounds on malondialdehyde formation induced by doxorubicin in rat liver mitochondria in vitro. AB - The isolated rat liver mitochondria were used in vitro to study the effect of doxorubicin on lipid peroxidation. We found that NADH-dependent mitochondrial peroxidation, measured by thiobarbituric acid (TBA) method, was stimulated to fourfold by doxorubicin (50 mumol.L-1). The addition of Fe3+ produced a significant increase of malondialdehyde (MDA) formation induced by doxorubicin. Doxorubicin enhanced the peroxidation of lipids in liver mitochondria through enzymatic mechanism especially in the presence of Fe3+. Probimane, bimolane, dexrazoxane (dioxopiperazine compounds), and edetic acid (EDTA) inhibited the formation of MDA in doxorubicin or doxorubicin+FeCl3 systems in a concentration dependent manner. The inhibitory rates of MDA formation by probimane at the concentrations of 0.01, 0.05, 0.1, and 0.25 mmol.L-1 were 27.80%, 25.19%, 47.80%, and 59.77% respectively; bimolane were 21.04%, 25.55%, 24.83%, and 54.13%; dexrazoxane were 11.29%, 20.68%, 34.94%, and 58.65%; EDTA were 57.52%, 55.67%, 61.62%, and 63.16% in Dox and FeCl3 system. The inhibitory rates of MDA formation by probimane at concentration 0.01, 0.05, 0.1, and 0.25 mmol.L-1 were 19.27, 39.02, 59.60, and 58.63% respectively; bimolane were 6.10, 17.19, 41.58, and 53.22%; dexrazoxane were 27.24, 33.26, 58.21, and 59.11%; EDTA were 63.76, 67.43, 61.68, and 63.27% respectively in Dox system. These results suggested that protection against cardiotoxicity of doxorubicin afforded by probimane, bimolane, and dexrazoxane may be related to their ability to combine with the complex iron so that the iron was no longer able to take part in free radical reactions. PMID- 8249631 TI - Protective effects of flunarizine on hemorrhagic shock in rats. AB - Intravenous injections of flunarizine 0.25, 0.5, or 1.0 mg.kg-1 10 min before hemorrhage increased the maximal bleeding volume from 4.3 +/- 1.1 to 5.5 +/- 1.1 ml. As the dose of flunarizine increased, the survival time in rats subjected to hemorrhage was prolonged in a dose-dependent manner. Five hours after the reinfusion, flunarizine 1 mg.kg-1 markedly improved the survival rate to 70% compared with nil in the shock group. Flunarizine reduced the increase of lactate in blood, ameliorated the depletion of ATP stores in tissues, and prevented the calcium accumulation in heart and liver. The results suggest that flunarizine may produce a protective effect on hemorrhagic shock, probably related to the decrease of calcium accumulation in the ischemic tissues. PMID- 8249632 TI - Effects of glaucocalyxin A on aggregation and cAMP levels of rabbit platelets in vitro. AB - Glaucocalyxin A (Gla A) is a new diterpenoid isolated from ethereal extract of the leaves of Rabdosia japonica (Burm f) Hara var glaucocalyx (Maxim) Hara (Labiatae) collected in Northeastern China. When incubated with washed rabbit platelets, Gla A inhibited ADP-, AA-, and PAF-induced aggregation of rabbit platelets with IC50 values of 3.44, 13.32, and 7.74 mumol.L-1, respectively. Gla A 10 and 100 mumol.L-1 increased the cAMP levels in platelets. In combination with imazodan hydrochloride, Gla A (1-100 mumol.L-1) caused a marked increase of platelet cAMP levels, while no effect with PGE1. PMID- 8249633 TI - [Image analysis of effects of 4 drugs on coronary occlusion and myocardial reperfusion injury]. AB - Myocardial infarction induced by coronary occlusion (O model) and reperfusion (R model) and the effects of nitroglycerin (Nit), propranolol (Pro), lidocaine (Lid), nifedipine (Nif), and saline were studied in rats by image analysis and weighing methods. The results showed that the myocardial infarct size (MIS) in R and O models were 27 +/- 8% and 39 +/- 6%, respectively (P < 0.01). The roundness and the distances from the border of the infarct zone (IZ) to endocardium or epicardium in R model were greater than those in O model, while the gray level difference between normal and infarct myocardium (delta G) in the R model was less than that in O model. MIS of Nit group in O and R models were 23 +/- 12% and 16 +/- 7%, respectively, which were significantly less than those in Lid, Nif groups, and saline. The reduction of MIS was also found in Pro group in both models. The results suggested that early restoration of blood flow resulted in the salvage of injured myocardium and Nit and Pro were found to have beneficial effects in both models. PMID- 8249634 TI - [Potentiating effect of clonidine on anxiolytic action of buspirone in rats]. AB - The anxiolytic effects of buspirone (Bus) and clonidine (Clo) were studied in the social interaction (SI) test and the elevated plus-maze (EPM) in rats. Bus 1 mg.kg-1 (ip, 15 min) and Clo 10 micrograms.kg-1 (ip, 15 min) prolonged the active interaction time (AIT) in the SI test. While neither Bus 0.3 mg.kg-1 nor Clo 3 micrograms.kg-1 was effective in this model, combination of them increased AIT. Yohimbine 30 micrograms.kg-1 (ip, 15 min before Bus) reversed such a potentiating effect. On the other hand. Bus 0.3 mg.kg-1 or/and Clo 3 micrograms.kg-1 did not affect the VI (variable interval) schedule-controlled behavior. In EPM test, neither Bus 0.5 mg.kg-1 nor Clo 30 micrograms.kg-1 increased the percentages of open-arms entries (OE%) or of the time spent in open-arms (OT%). But these 2 indices were increased by combining Bus 0.5 mg.kg-1 with Clo 30 micrograms.kg-1. The results suggested that Clo potentiated the anxiolytic effect of Bus probably by its alpha 2-adrenoceptor agonist action in SI and EPM tests. PMID- 8249635 TI - [Protective effect of panaxatriols on function of reproductive endocrine axis in radiation-injured rats]. AB - After whole body irradiation with X-ray 5 Gy (radiation condition: 0.31 Gy.min-1, 200 kV, 10 mA, 0.5 mm Cu and 1.0 mm Al), the male rats were given ip panaxatriols 5 mg.d-1 24 h before and after irradiation for 14 d. The results showed that hypothalamic leu-enkephalin (165 +/- 12 vs 131 +/- 14 pg.mg-1), pituitary beta endorphin (2.3 +/- 0.5 vs 1.6 +/- 0.3 ng.mg-1) contents, and serum testosterone (1.66 +/- 0.15 vs 0.82 +/- 0.23 ng.ml-1) level were decreased, while serum FSH (1.34 +/- 0.10 vs 1.99 +/- 0.10 ng.ml-1) level increased in irradiation group vs normal control. These indices approached to control levels in irradiation + panaxatriols group. These suggest that panaxatriols have protective effects on reproductive endocrine axis and promote their recovering course from the radiation injuries. PMID- 8249636 TI - [Sodium channel blocking effect of scorpion venom on cultured mouse myocardiocytes]. AB - Myocardiocytes of mice were cultured. Action potentials were recorded with microelectrodes inside the cells. Scorpion venom from Buthus martensii Karsch 3.75 or 7.5 micrograms.ml-1 decreased the duration of action potential and all of the depolarization concerned parameters of myocardiocytes. The Vmax, TP, APA behaved apparently in a dosage-dependent way. Restoration happened after washing out. Tetrodotoxin 2.5 micrograms.ml-1 acted in a similar way. Nimodipine 3.0 micrograms.ml-1 led to a decrease in action potential duration. BaCl2 0.1 mmol.L 1 elongated the action potential duration, while decreased the parameters concerned with depolarization. These results indicate that the scorpion venom has Na+ channel blocking action. PMID- 8249637 TI - [Effects of oxyfedrine on high blood viscosity and myocardial necrosis induced by epinephrine and ice water stress in rats]. AB - Acute high blood viscosity (HBV) and myocardial necrosis was established by epinephrine (Epi) and ice water stress in rats. Effects of iv oxyfedrine (Oxy) on HBV, plasma viscosity (PV), hematocrit, erythrocyte electrophoretic time (EET), and fibrinogenic viscosity (FV) were studied in model. Results showed that Oxy 1 mg.kg-1 iv markedly decreased the arterial and venous blood HBV at shear rates of 700 s-1 and 70 s-1, respectively (P < 0.01). There were significant differences in the alleviation of HBV among 3 groups (Oxy 0.01, 0.1, and 1 mg.kg-1 iv). The above doses markedly decreased the HBV, PV, and FV, and shortened the EET. Effects of iv Oxy on the myocardial necrosis rat model were scrutinized under the light and electron microscopes. Oxy iv 1 mg.kg-1.d-1 x 1, 3, and 5 d prevented or mitigated the occurrence and development of myocardial necrosis. The structure of heart mitochondria and myofibrils were clearly discernible. This action may be related to the alleviation of HBV by Oxy. PMID- 8249638 TI - [Protective effects of 3, 6-dimethamidodibenzopyriodinium citrate on myocardial injury induced by ischemia and reperfusion in rats]. AB - 3, 6-Dimethamidodibenzopyriodonium citrate (I-65) 0.5 and 1 mg.kg-1 pretreatment reduced the size of myocardial infarct after ischemia for 40 min and reperfusion for 120 min. I-65 0.5 mg.kg-1 decreased myocardial creatine kinase, lactate dehydrogenase release, and Ca2+ accumulation after ischemia for 40 min and reperfusion for 120 min. The results show that I-65 prevents cardiac ischemia and reperfusion injury and the effect is considered to be related to inhibition of myocardial Ca2+ accumulation. PMID- 8249639 TI - [Effects of preimplantation treatment with aspirin and acetaminophen on blastocyst and fetus in rats]. AB - Pregnant rats were treated with ig aspirin (Asp) and acetaminophen (Ace) on d 3 of pregnancy (positive vaginal smear = d 0). Blastocysts were collected on d 4 and evaluated for gross morphology, cell number, micronucleus, and mitotic index. Some rats were killed on d 20 and fetuses were examined for teratogenic effects. On d 4 a reduction of cell number per blastocyst was found in the rats treated with Asp 0.5, 1 g.kg-1, and Ace 1 g.kg-1, while the mitotic index, frequency of micronuclei, and frequency of blastocysts with morphological alterations were increased. The frequency of micronuclei was increased in rats exposed to Ace 0.25 and 0.5 g.kg-1. On d 20 major malformation and embryotoxicity were seen in Asp 0.5, 1, and Ace 1 g.kg-1 groups. PMID- 8249640 TI - [Effects of clonidine on estrus, estradiol, gonadotropin, graafian follicle, and corpus luteum in rats]. AB - Effects of clonidine (Clo) on female reproductive system were studied in rats. Blood FSH, LH, progesterone, testosterone, and estradiol were measured by radioimmunoassay and the development of secondary follicles and corpus luteum in ovary were investigated by morphometry. After Clo po 0.3 mg.kg-1.d-1 x 14 d, the estrus of rats was prolonged; FSH, LH, and progesterone increased significantly; while estradiol reduced. The development of secondary follicles in ovary was blocked at the stage of prematuration and the numerical density of corpus luteum decreased. After clonidine po 28 d, FSH and LH sustained at high levels, but the estrous cycle, estradiol and progesterone recovered. PMID- 8249641 TI - [Protective action of blumeatin against experimental liver injuries]. AB - Blumeatin (Blu, 5,3',5'-trihydroxy-7-methoxy-dihydro-flavone) was first isolated from Blumea balsamifera DC by Department of Chemistry, Sunyatsen University of China. Blu ip inhibited the increase of serum alanine aminotransferase (AAT) and liver triglyceride and increased serum triglyceride, beta-lipoprotein, and liver glycogen content in CCl4-intoxicated rats. Histological lesions of liver were less severe than those of hepatic injury control. Blu ip 0.65 and 3.25 mg.kg-1 inhibited the increase of serum AAT and hepatic TG in thioacetamide (TAA) intoxicated mice. Blu ip shortened the pentobarbital sleeping time in CCl4 intoxicated mice. It suggested that Blu could protect liver against injury induced by CCl4 and TAA. PMID- 8249642 TI - [Effect of thymic factor D on lipid peroxide, glutathione, and membrane fluidity in liver of aged rats]. AB - The effects of thymic factor D (TFD) on aging were investigated in male Wistar rats at the age of 24 months. Thymic factor, polypeptides extracted from swine thymus, was given to aged rats in the dose of 2 mg.kg-1 sc every other day for 3 months. It was shown that TFU decreased the content of malondialdehyde (MDA) and lipofuscin in the liver of aged rats, accompanied by an increase of glutathione (GSH). Besides, the declined fluidity of hepatic mitochondrial and microsomal membranes in aged rats was recovered to the level of young adult rats by administration of TFD. It is suggested that TFD may possess beneficial effects on retarding the senile metabolic changes in hepatic cells. PMID- 8249643 TI - Post-dexamethasone cortisol levels and computerized tomographic findings in manic patients. AB - The relations of post-dexamethasone cortisol levels to computerized tomography (CT) scan findings of lateral and third ventricle size and cortical sulcal prominence rated separately for frontal and parieto-occipital distributions were examined in 15 manic patients. Failure to suppress endogenous cortisol after a dexamethasone challenge was associated with larger third ventricles, but not to any of the other CT scan measures. PMID- 8249644 TI - Premenstrual changes: are they familial? AB - To investigate the potential familial component in premenstrual changes, 80 pairs of sisters were assessed with the Premenstrual Assessment Form, a retrospective measure, and the Daily Ratings Form, a prospective measure, for physical, behavioral and mood changes during the premenstrual phase of the menstrual cycle. Sisters were not found to be concordant for premenstrual changes on measures derived from either procedure, a finding contradictory to previous reports in the literature as well as to our own prior research. PMID- 8249645 TI - Seasonality in severity of depression: relationships to suicide and homicide occurrence. AB - Some previous studies have reported seasonal or monthly variations in the occurrence of depressive syndromes. The present study was carried out in order to investigate seasonality in severity of depression. Toward this end, the authors measured the Zung Self-Rating Depression (ZD) and Anxiety (ZA) Scales scores in 104 consecutively admitted depressed patients between November 1983 and April 1985. The data were analyzed by means of spectral analysis of a single time series. Up to 47.9% of the variance in the weekly average of the ZD scores could be explained by two significant rhythms of 51 (circannual) and 7 weeks. Peaks in ZD scores were observed in April-May, with lows occurring in August-September. Up to 30.8% of the variance in the weekly average of ZA scores was explained by a circannual rhythm. Our results show that there is a true seasonality in the severity of illness of depressed subjects. There were significant correlations between the weekly average in severity of illness and the chronograms of suicide (positively) and homicide (negatively) occurrence in Belgium. PMID- 8249646 TI - Vietnamese boat refugees: the influence of war and flight traumatization on mental health on arrival in the country of resettlement. A community cohort study of Vietnamese refugees in Norway. AB - A consecutive cohort of 145 adult Vietnamese refugees were personally interviewed and completed the Symptom Checklist 90 R self-rating scale on arrival in Norway. Sixty-two percent had witnessed bombing, fires and shooting, 48% had witnessed other people being wounded or killed and 36% had been involved in life threatening situations or had been wounded in the war. Nearly all war trauma variables but none of the escape or refugee camp variables were significantly related to mental health 7 years after the end of the war. War trauma was significantly associated with mental health, also when age, gender and previous mental problems were controlled for. These results and our clinical experience indicate that clinicians treating refugees should address such traumatic experiences specifically. PMID- 8249647 TI - Short-term and intermediate-term outcome in adolescent eating disorders. AB - Within a prospective longitudinal outcome study of the eating disorders with onset during adolescence, 26 former patients were reassessed at short-term (19-38 months) and intermediate-term (48-72 months) follow-up periods. It was shown that the outcome was worse at short-term follow-up. In addition, the changes of diagnostic categories within the spectrum of the eating disorders were studied. It is concluded that outcome studies require repeated assessments to study the various patterns of course of the eating disorders. PMID- 8249648 TI - Predicting the long-term outcome of affective disorders. AB - Factors influencing the long-term outcome of affective disorders were investigated in 106 patients with an average length of course of 28 years. Univariate statistical analyses, stepwise multiple discriminance analyses and path analysis were applied. A direct unfavourable influence on the frequency of persisting alteration (evaluated according to the Global Assessment Scale) was found to have the low self-confident premorbid personality and a higher number of episodes during course, whereas only the variable "male" had such an influence on the social consequences of the illness. The variable "manic episode" (bipolarity) and "age at onset" had only an indirect influence, in that bipolar patients had more episodes, and younger patients more frequently had a bipolar course and therefore had more episodes. The unfavourable long-term outcome was due in those cases to the higher number of episodes. PMID- 8249649 TI - Redefining epidemic hysteria: an example from Sweden. AB - Francois Sirois' influential paradigm for diagnosing episodes of epidemic hysteria is discussed. The part of his schema addressing the large diffuse outbreak should be eliminated as it does not possess characteristic features of mass hysteria and overlooks the potential social, cultural, political, ritualistic and institutional patterning of collective behavior. A case study involving the collective delusion of phantom rockets over Sweden during 1946 illustrates the complexities of such episodes. PMID- 8249650 TI - Personality disorders in DSM-III-R as categorical or dimensional. AB - Despite the atheoretical approach of the DSM-III-R, the personality disorders have their roots in specific theoretical schools. Due to clinical tradition, analogies with Axis I and the tradition in psychiatry and medicine, the personality disorders are presented as categories although there is more empirical support for a dimensional approach. This study attempted to determine whether the separate personality disorders meet Kendell's criteria for distinct entities, i.e., bimodality with distinct points of rarity. None of the personality disorders met the criteria for a distinct disease entity. Instead, all of the personality disorders presented as continuous, dimensional personality traits present among healthy subjects and more pronounced in patients with mental disorders. PMID- 8249651 TI - The test-retest reliability of the Mini-Mental State Examination in chronic schizophrenic patients. AB - The test-retest reliability of the Mini-Mental State Examination (MMSE) was explored in a sample of 22 inpatients suffering from chronic schizophrenia by DSM III-R criteria. Patients were tested 4 times during a period of 4-6 weeks. The total score for serial sevens showed consistently significant reliability at about 0.7, and the total score for spelling displayed a lower reliability at about 0.4, which was of borderline significance. In this short period of time, 36% and 27% of the patients shifted between being demented and nondemented when the serial sevens and spelling, respectively, were used to calculate total scores. The MMSE should not be used to categorize schizophrenics as demented or nondemented in biological studies using relatively stable variables. PMID- 8249652 TI - Parental rearing and intimate relations in women's depression. AB - Bowlby has suggested that attachment behavior is not restricted to early childhood but can remain valid through the life span. This study was designed to test whether recall of parental rearing (Parental Bonding Instrument) and perception of marital relationship (Dyadic Adjustment Scale) is significantly different between 2 groups of women: one with non-bipolar depressive disorder (DSM-III-R) compared with another (control) of healthy women from a primary practice setting. We also examined the hypothesis that exposure to dysfunctional parenting is associated with negative intimate relationships in adulthood. Our results partially support these hypotheses. We discuss the significance of these findings in the prevention and treatment of depressive disorders. PMID- 8249653 TI - Mianserin and chronic pain: a double-blind placebo-controlled process and outcome study. AB - There is evidence that antidepressants may have an analgesic effect in chronic pain. To replicate this effect and to throw light on the processes involved, a 12 week cross-over double-blind trial of mianserin versus placebo was carried out in 4 diagnostic groups: A) depressive patients without pain complaints (n = 8), B) depressive patients with chronic organic pain (n = 8), C) patients with somatoform pain disorder and vital signs of depression (n = 11) and D) patients with chronic organic pain without depression (n = 8). Although a mianserin induced antidepressant effect in group A was evident, no significant pain reduction was accomplished in any group. The reasons for this failure to replicate the antidepressant-induced analgesic effect are discussed. PMID- 8249654 TI - Sex differences in depressiogenic risk factors and coping strategies in a socially homogeneous group. AB - We examined for sex differences in possible risk factors for depression, manifestations of depression and associated help-seeking and coping styles in a socially homogeneous cohort without any sex difference in lifetime rates of depression. The sexes did not differ on measures of depression, dysfunctional attitudes, perception of important interpersonal relationships, experience or perceived impact of life events. Women scored more highly on measures of neuroticism, were more dependent and were more likely to engage in self-consoling behaviours and coping styles. Recklessness as a coping style was the only item endorsed more frequently by men, posing the question of whether current measures take into consideration the male experience of depression. The findings suggest that many putative depressiogenic risk factors over-represented in women do not, by themselves, create depression but require a triggering factor or certain social conditions for their expression. PMID- 8249655 TI - Obsessive-compulsive disorder in children and adolescents. Self-reported obsessive-compulsive behaviour in pupils in Denmark. AB - A total of 1032 pupils in Denmark aged 11-17 years completed a survey form of the Leyton Obsessional Inventory - child version, consisting of 20 items. These pupils had higher scores for obsessional symptoms or traits than high school students in the United States, also when corrected for age differences, but the most frequently reported symptoms seemed to be the same: repetitive thoughts or words, having trouble making up one's mind and worrying about being clean enough. There was no significant sex difference in total score. From 0.5% to 5% reported considerable interference by each obsessional symptom, and 4.1% had a total interference score > 25, reflecting possible subclinical or clinical obsessive compulsive disorder. PMID- 8249656 TI - Serum thyrotropin responses to thyrotropin-releasing hormone in Korsakoff's syndrome. AB - Eleven patients with DSM-III-R Korsakoff's syndrome were studied as were 11 patients with alcohol dependence syndrome (without Korsakoff's syndrome and abstinent for at least 3 years) and 11 healthy controls. All subjects underwent a thyroid-releasing hormone and thyroid-stimulating hormone stimulation test. Forty five percent of the Korsakoff's syndrome patients had a blunted response and 18% of the chronic abstinent alcoholic group were similarly blunted. None of the controls had an attenuated response. The possible reasons for the high level of blunting are discussed. PMID- 8249657 TI - Severity of family distress of schizophrenia--fact or artifact? PMID- 8249658 TI - Suicide in the military as a function of involvement in war. PMID- 8249659 TI - Effects of early undernutrition on kinetic parameters of brain acetylcholinesterase from adult rats. AB - The effects of undernutrition during early life on acetylcholinesterase (AChE) specific activity of cerebellum, hippocampus, hypothalamus and striatum were examined in rehabilitated adults rats. Undernourished rats were raised by dams maintained on a restricted food scheme from pup birth to pup weaning (day 31). The offspring were maintained on a restricted food schedule until day 38 when they started to have free access to food until 70-80 days of age. Control rats were raised by dams which had free access to food. The results showed that early malnutrition caused a significant increase in AChE specific activity in cerebellum (about 20%), striatum (about 40%), and hypothalamus (about 30%). No changes were found in the hippocampus. Undernutrition caused a significant increase in Vmax when compared to the control group without changes in Km both in cerebellum and striatum. These results suggest that early undernutrition changes AChE concentration in cerebellum and striatum and does not affect the affinity of the enzyme for the substrate. PMID- 8249660 TI - Behavioural changes after ablation of subdivisions of the rat prefrontal cortex. AB - Sham operated controls and four groups of rats subjected to ablation of various parts of their frontal cortex were compared in food and water intake and four behavioural tasks. The ablations were aimed at removing (1) the ventral prefrontal cortex, (2) the dorsal part of the medial prefrontal cortex, (3) the total medial prefrontal cortex, and (4) the anterior dorsolateral (non prefrontal) cortex. Only two groups had a significantly impaired food and water intake: the ventral prefrontal and the non-prefrontal anterolateral. The latter group was not adipsic. Two variants of spontaneous alternation were administered in a T-maze: a non-cued version in which both arms were grey and a cued version in which one arm was white and the other black. While all ablated groups behaved like the control group on the non-cued test, the number of perseverative responses of the total anteromedial group was significantly increased in the cued version of the test. Significant group differences could be seen neither in a test of conditioned taste aversion nor in extinction of operantly conditioned bar presses. Finally, in a vertical hole-board exploration test the only significant group difference was a prolongation of the mean visit duration of the ventrally lesioned animals in comparison with all other groups. The results of the present study further indicate functional differentiation within the prefrontal cortex of the rat. PMID- 8249661 TI - The role of sexual reward in the temporal patterning of copulatory behaviour in male rats. AB - In 8 male rats instrumental responses reinforced by contact with an estrous female with an intact or a surgically closed vagina were investigated. Contact with the female lasted until the end of the mount bout. It was observed that neither the rewarding value of the mount bout (expressed by the duration of the response) nor its arousing properties (expressed by the rate of bar presses) depended on the quality (mounts or intromissions) or quantity of the copulatory events of which it was composed. It is suggested that the temporal patterning of copulatory behaviour in male rats depends on fluctuations of sexual reward. This sexual reward can be a result of both genital stimulation as well as the performance of mounting under the influence of sign stimuli coming from the estrous female. PMID- 8249662 TI - Dilatation of the lateral part of the transverse fissure of the brain in Alzheimer's disease. AB - Post-mortem MRI (magnetic resonance images) studies followed by histopathological examination were used to study the size and the shape of the lateral part of the transverse fissure of the brain in seven individuals with Alzheimer disease (AD) and five controls. In control brains, the lateral part of the transverse fissure is a narrow cleft protruding laterally as choroid and hippocampal recesses. In AD affected brains, the lateral part of the transverse fissure becomes a large subarachnoid space as a result of different degrees of atrophy of various hippocampal and parahippocampal structures. Our findings directly indicate the relationship between changes in the hippocampal and parahippocampal structures and the size of the lateral part of the transverse fissure. Sector CA1, the subiculum, the entorhinal cortex, and the parahippocampal isocortex are the most affected, whereas the dentate gyrus is much less affected. Adjacent thalamic structures, which are less vulnerable to the AD pathology, do not appear to contribute to transverse fissure changes. The size and the shape of the lateral part of the transverse fissure of the brain in AD reflect the atrophy of the hippocampus and parahippocampal structures. PMID- 8249663 TI - Learning performance at different time periods after hypoxia in infant rats. AB - Infant rats were exposed to hypoxia. At different time periods after the hypoxic aggression the animals were subjected to a one-way active avoidance task consisting of 3 daily training sessions. Learning performance of hypoxic rats, expressed as the number of avoidances per 10 trials, was significantly decreased in all the sessions at posthypoxic day 35 (P < 0.001). The rate to which learning behaviour was affected by hypoxia was comparable among all 3 sessions in both groups. Histological evaluation of hippocampus and cortex at day 7 and day 35 after hypoxic exposure, did not reveal detectable structural changes. It can be concluded from this study that hypoxia, induced in 21 day old rats leads to learning performance deficits in the adult animal. PMID- 8249664 TI - Evidence for a D4 dopamine receptor decreasing serotonin N-acetyltransferase activity in chick retina. PMID- 8249665 TI - Salt appetite: its neuroendocrine basis. AB - Based on the early work of Richter (1936), showing that the adrenalectomized rat kept itself alive by drinking hypertonic NaCl solutions, Epstein and Stellar (1955) demonstrated that that salt appetite was innate, not dependent on learning. A series of later papers by Epstein and his students made clear that in addition to the adrenal steroid, aldosterone, salt appetite depended upon the action of angiotensin II in the brain. Blocking either hormone in the brain reduced depletion-induced salt appetite in half; blocking both eliminated it. Two salt depletions enhanced depletion-induced salt appetite by nearly a factor of two even when the rats never had a chance to drink salt in the first depletion. With multiple depletions, need-free salt intake also increased when the rats were sodium replete, producing an chronic, elevated salt appetite. Strikingly, female rats drink almost twice as much as males and become more enhanced by prior depletions. The neural circuitry involved in the synergy of angiotensin and aldosterone is becoming clearer with lesions of the amygdala that reduce aldosterone's effects and lesions of the anterior wall of the third ventricle that reduce angiotensin's effects. The significance of salt appetite in nature, in body fluid homeostasis, and as a model system of the brain mechanisms of ingestive behavior is discussed. PMID- 8249666 TI - Social effects on development of food preferences. AB - The purpose of this study was to answer the question as to whether the food preferences of the young are dependent on the social influences or not. The experiments were conducted on female cats bearing electrodes implanted in the mid lateral hypothalamus, and their weanling kittens. Each mother was given a self stimulation test; the mothers which learned to press a lever for the hypothalamic stimulation reward were chosen for the experiment. Each mother, always 4 h food deprived before the session, was given a choice of meat pellets and banana slices. Eating bananas was rewarded by the hypothalamic stimulation whereas eating meat pellets was not; as a result, the mother concentrated only on eating bananas while ignoring meat pellets. In the following sessions one or two weanling kittens were always accompanying the mother during the session. It was found that 15 out of 18 weanling kittens used in this experiment joined the mothers in eating bananas. After separation from the mothers these kittens continued to choose bananas and ignore meat pellets when tested in the absence of the mother. Control kittens of the same age, which were never trained with the mother, refused to eat bananas. Similar results were obtained with other mother kittens groups in which mashed potatoes or plain jellied agar were given instead of bananas. These results suggested that the food preferences of the weanling kittens were influenced by the mother's choice of food, even in the case when this food was unusual for the species. PMID- 8249667 TI - Patterns of line bisection in patients suffering from unilateral spatial neglect. AB - Thirteen patients suffering from Unilateral Spatial Neglect and 6 Right Brain Damaged Control Patients were tested on a line bisection task in order to verify peculiar patterns of error. Stimuli were arranged in order to avoid confounding the effects of line length and line position in the space. Two parameters of rightward displacement of setting point were used: 1) the distance of patient's setting point from actual line midpoint, and 2) the deduced left end-point of the line that patient took into account. Results showed that: 1) the rightward extension of lines did not lead to a consistent rightward displacement of setting point; 2) error significantly increased as lines extended more than 10 cm in the left hemispace; 3) the USN severity amplified the degradation of leftmost portion of stimuli until the deduced left endpoint of all space/length conditions was aligned on the left of patient's sagittal midplane. The interpretation of these results points to the particular arrangement of neurons directing attention on specific portions of the visual field. PMID- 8249668 TI - The presence of anti-Borrelia burgdorferi antibodies in a group of multiple sclerosis patients in eastern Sicily. Preliminary data. AB - The authors evaluate the presence of anti-Borrelia burgdorferi antibodies in a group of polysclerotic patients of Eastern Sicily, in order to verify or dismiss a correlation between Borrelia infection and demyelinizing syndrome. 460 samples of serum were examined, of which 60 belonged to patients affected by definite multiple sclerosis; the other 400 were taken from a group of apparently healthy subjects. From the analysis of the data obtained it can be seen that of the 60 polysclerotic subjects examined, 12 equal to 20% were found to be positive for the presence of IgG anti-Borrelia burgdorferi, while in the control group 30 subjects were found to be positive, equal 7.5%. PMID- 8249669 TI - Acid maltase deficiency in childhood. Early diagnosis and clinical follow-up of late-onset glycogen storage disease type II. AB - A case is described of late-onset glycogenosis type II presenting with an isolated rise in serum transaminase levels. Histological, histochemical, ultrastructural and biochemical examinations performed on muscle biopsy showed the typical laboratory features of late-onset glycogenosis type II, which was diagnosed more than four years before the first appearance of disease-related signs and symptoms. A heterozygote status for the same defect was also demonstrated by enzyme assays in both parents, thus confirming the autosomal recessive mode of inheritance of the disorder. Even though an elevation in transaminases and other serum enzymes of possible muscle origin has been previously described as a diagnostic clue in some unsuspected muscular diseases in childhood, as far as we know no other patient with a sporadic form of glycogenosis type II has been identified when still completely asymptomatic. The possibility of silent primary metabolic diseases and myopathies should be carefully considered when evaluating children with persistently elevated serum transaminases, even in the absence of suggestive anamnestic, familial and physical findings, in order to obtain an early diagnosis and to provide an appropriate genetic counselling. PMID- 8249670 TI - Paroxystic neuropsychological symptoms as the early expression of hepatic encephalopathy. A case report. AB - A 52-year-old patient presented with paroxystic episodes of generalized apraxia, anomia, agraphia and acalculia. The transient character of these attacks was supported by several neuropsychological examinations. Initially a tentative diagnosis of multiple TIA's was made. Treatment consisted of antiplatelet aggregation therapy. Three years later, however, paroxystic neuropsychological symptomatology occurred more frequently with an increase of severity. The patient was again seen and the differential diagnosis included epilepsy or a metabolic disturbance, in casu hepatic encephalopathy. A therapeutic trial with carbamazepine was started but the patient deteriorated further. He developed a flapping tremor and became stuporous. The blood ammonia was high and there were triphasic waves on the EEG. A probable diagnosis of hepatic encephalopathy was made and carbamazepine therapy was withdrawn. There was a good response on low protein diet and lactulose. PMID- 8249671 TI - Thymectomy for myasthenia gravis: predictive factors and long term evolution. A retrospective study on 46 patients. AB - Forty-six patients with myasthenia gravis (MG) underwent transternal thymectomy between 1975 and 1991 and were observed over a long term follow up period (mean 6 years). Surgery was well tolerated and was followed in 32.6% of patients by remission of symptomatology. The severity of symptoms according to the Osserman scale was significantly reduced: slight forms of myasthenia increased soon after thymectomy more frequently than moderate and severe forms. Response to thymectomy at 3 months, according to the Hankins scale, was found to be positive (remission or amelioration of symptoms) in 31/41 patients, and worse in patients with thymoma. Probability of clinical remission and death were both significantly correlated with clinical response at three months after thymectomy. No other preoperatory clinical factor (onset age, sex, initial severity of MG, interval from symptom onset to thymectomy) was found to predict the response to treatment. Preoperatory mediastinic CT had a good reliability (> 85%) in diagnosing thymona, but was rarely able to distinguish between thymic hyperplasia and atrophy. PMID- 8249672 TI - Transient "cerebellar" mutism in lesions of the mesencephalic-cerebellar region. AB - Four patients aged from 20 to 48 years with transient mutism are presented: 3 patients underwent surgery for midline tumours of the mesencephalic-cerebellar region (medulloblastoma in two cases and pinealoblastoma in one), at times attached to one or both lateral recesses of the IV ventricle. One patient was hospitalized and treated for brain-stem ischemia. All patients developed mutism 48 to 72 hours after surgery; in the patient with brain-stem softening mutism appeared 72 hours after admission. All the patients had unimpaired consciousness and no deficits of lower cranial nerves. Speech, always normal in the first hours after surgery, was regained after a period of 6-16 weeks. Various hypotheses for this speech disorder are analyzed. PMID- 8249673 TI - Multiple cerebral infarctions in a young patient with secondary thrombocythemia due to iron deficiency anemia. AB - A 30-year-old woman developed multiple cerebral infarctions. In the absence of other risk factors, thrombocythemia secondary to iron deficiency anemia due to polymenorrhoea was considered to underlie the cerebral infarctions. Platelet count was normalized after iron therapy. The importance of vigorous treatment of iron deficiency anemia in preventing complications of secondary thrombocythemia is emphasized. PMID- 8249674 TI - Antiphospholipid antibodies and cerebrovascular disease. AB - Antiphospholipid antibodies (lupus anticoagulant and anticardiolipin antibodies) are regarded as important risk factors for ischemic stroke and transient ischemic attacks in young subjects. In fact the interaction of these antibodies with phospholipid may impair the coagulation system at several steps, promoting thrombosis. A variety of therapeutic strategies including corticosteroids, immunosuppressive drugs, antiplatelet agents and anticoagulants have been prescribed in stroke patients with aPLs, but the efficacy of these drugs has not been established yet. PMID- 8249675 TI - Impaired glucose-induced insulin secretion: studies in animal models with spontaneous NIDDM. PMID- 8249676 TI - Molecular genetics of NIDDM and the genes for insulin and insulin receptor. PMID- 8249677 TI - Autoregulation of glucose transport: effects of glucose on glucose transporter expression and cellular location in muscle. PMID- 8249678 TI - Insulin resistance and the pathogenesis of non-insulin dependent diabetes mellitus: cellular and molecular mechanisms. PMID- 8249679 TI - Perturbation of islet metabolism and insulin release in NIDDM. PMID- 8249680 TI - Indirect effects of insulin in regulating glucose fluxes. AB - Metabolism of fuels is driven by the energy demand of the organism and its regulation is influenced by many hormonal and metabolic factors. Insulin is of utmost importance in regulating glucose metabolism by promoting glucose uptake in the insulin-sensitive tissues for energy consumption and/or storage. The effects of insulin on glucose metabolism can be both direct and indirect. Ample evidence has indicated that insulin directly stimulates glucose transport systems in the target tissues. However, the changes in glucose fluxes can also be brought out by indirect effects of insulin which are produced secondary to the insulin-induced changes in other hormones and metabolites. In this chapter, we discussed a number of examples of insulin's indirect effects on glucose metabolism. We demonstrated that insulin can indirectly promote muscle glucose uptake during exercise by restraining the release and oxidation of fatty acids and decrease of hyperglycemia. We have presented some evidence for an indirect regulation of glucose cycling by insulin. We have also demonstrated the importance of the peripheral levels of insulin for insulin-induced inhibition of hepatic glucose production. This presumably indirect effects of peripheral insulin might consist of 1) suppression of the release of energy substrates and gluconeogenic precursors; and 2) suppression of glucagon secretion. In a carbachol-induced stress model, insulin is not required for a putatively neural regulation of an increase in systemic glucose uptake but a "permissive" effect of insulin is essential. These studies underscore the importance of the interactions between insulin and other hormones and metabolites as opposed to insulin's direct actions per se. PMID- 8249681 TI - The insulin-antagonistic effect of the counterregulatory hormones--clinical and mechanistic aspects. PMID- 8249682 TI - On insulin action in vivo: the single gateway hypothesis. PMID- 8249683 TI - Acute hormonal regulation of gluconeogenesis in the conscious dog. PMID- 8249684 TI - Estimating gluconeogenic rates in NIDDM. AB - To measure the rate of gluconeogenesis in humans directly, one must administer and determine the specific activity or the enrichment in an intermediate in the gluconeogenic process and in the glucose formed, thus obtaining the fraction of the glucose formed by gluconeogenesis. By a separate determination of the rate of hepatic glucose production, the rate of gluconeogenesis can then be calculated. The closer the intermediate is to glucose-6-P, the more complete will be the measurement of the rate. Thus, if the intermediate is below the level of the triose phosphates, gluconeogenesis from glycerol will not be included in the estimate. Estimates of rates of gluconeogenesis from estimates of PEP enrichment or specific activity require a measure of the extent of exchange of label at the level of oxaloacetate. By using 14C or 13C labeled CO2 as the intermediate and estimating the relative rates of the reactions of the tricarboxylic acid cycle relative to gluconeogenesis from the distribution of 14C from [3-14]lactate in glutamine from the glutamine conjugate of phenylacetate, the enrichment or specific activity of PEP has been estimated. Correction must be made for the incorporation into the glutamine of 14CO2 formed from the [3-14C]lactate. Data support the validity of this approach toward estimating gluconeogenesis in NIDDM, but the approach is complex, time consuming and with uncertainties. Estimates that have been made using [2-14C] acetate are invalid because of the extensive metabolism of [2-14C]acetate in other than liver. Other approaches have promise, but technical problems may exist in their use and other problems, such as hepatic zonation and exchange reactions, may compromise their application. PMID- 8249685 TI - Glucose metabolism during physical exercise in patients with noninsulin-dependent (type II) diabetes. PMID- 8249686 TI - Substrates and the regulation of hepatic glycogen metabolism. AB - Glycogen metabolism is a complex process which depends on the metabolic circumstances and the hormonal milieu. In this overview an intriguing new possibility has been emphasized--the possible central role of lactate in coordinating, with glucose, the net synthesis of glycogen. Since lactate changes acutely under many physiological circumstances, it would be a logical candidate for a signal which communicates to the liver the metabolic states of the periphery. It would then acutely determine the synthetic rate of glycogen synthesis within the range determined by the glucose concentrations which in turn could be said to reflect the nutritional state of the system. Interestingly, after oral glucose loading, portal glucose levels would be about 25% higher (Radziuk et al., 1978) relative to arterial. As seen from Figs 8 and 9 however the glycogen synthetic rate appears very sensitive to glucose (at a given lactate uptake). Everything else being assumed equal therefore, more glycogen would be synthesized than during intravenous loading with an equivalent peripheral concentration. This is indeed the case (Shulman and Rossetti, 1989). On the other hand, during equivalent loads, peripheral glucose levels are higher and the same quantity of glycogen is synthesized (Radziuk, 1989a, 1989b). If lactate is typical of other glucogenic substrates, then it is also logical that mixed meals with higher levels of portal substrate would maximize glycogen synthetic rates. Similarly, in diabetes where hyperglycemia and hyperlactatemia prevail, gluconeogenesis plays a predominant role in glycogen synthesis (Giaccari and Rossetti, 1992).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249687 TI - Regulation of cytoplasmic free Ca2+ in insulin-secreting cells. PMID- 8249688 TI - Gluconeogenesis in type 2 diabetes. PMID- 8249689 TI - Regulation of adipose tissue lipolysis, importance for the metabolic syndrome. PMID- 8249690 TI - Cellular and molecular factors in adipose tissue growth and obesity. AB - Heparin-binding growth factors related to basic fibroblast growth factor are major determinants of the cellular clonal composition of adipose tissue. By providing and maintaining varying complements of preadipocytes in different fat depots, these factors contribute to the varying sizes and functions of different regions, including the hypercellularity in appreciable obesity. Thus, differing levels and activities of the heparin-binding growth factors contribute to variations in depots within the same individual and between individuals, in lean and obese states. In contrast to regional differences in adiposity, which are accounted by factors resident in adipose tissue, we believe that obesity results from a generalized energy overload. According to our concept, there are genetic variations in cytoskeletal activity and thus differing quantities of energy are utilized for biomechanical processes. In a reciprocal relationship, the higher the cytoskeletal activity, the lesser the energy available for chemical energy storage, mainly in the form of triglyceride in adipocytes. At the extreme of "supermassive" obesity, a mutation in a gene related to a cytoskeletal protein would lead to appreciable dampening of cytoskeletal activity, with consequently the greatest quantity of energy remaining available for eventual triglyceride storage. Moreover, the new concept, for which we have have increasing experimental evidence, invokes a hypothalamic-efferent neural-cytoskeletal pathway, which would modulate the activity of the cytoskeleton. PMID- 8249691 TI - Regional obesity and NIDDM. AB - Obesity and NIDDM are clearly linked. The subgroup of abdominal, visceral obesity has been shown to have a particularly close link to the development of diabetes. This is probably due to the marked insulin resistance of that condition. Epidemiological data show a predictive power for the development of NIDDM in both sexes, in signs of insulin resistance, visceral obesity and, in women, hyperandrogenicity. In men a relative hypogonadism may be of importance. Experimental evidence suggests cause-effect relationships between these factors. In both sexes visceral fat may contribute to insulin resistance in the liver and the periphery by excess production of FFA. Hyperandrogenicity in women may also cause insulin resistance, although the reverse sequence of events cannot be excluded. The relative hypogonadism may well contribute to insulin resistance in men, as well as to the accumulation of visceral fat. There are observations of additional endocrine aberrations in visceral obesity, suggesting a central, neuroendocrine disturbance, which might be a primary factor for the pathogenesis of the syndrome. PMID- 8249692 TI - Hyperinsulinemia and VLDL kinetics. PMID- 8249693 TI - Hyperinsulinism and dyslipidemias as coronary heart disease risk factors in NIDDM. AB - In conclusion hypertriglyceridemia is accompanied by multiple metabolic disturbances which are potentially atherogenic. Atherogenic pattern of risk factors in insulin resistance syndrome request early intervention to prevent the development of CHD. The practical implication is that the presence of cardiovascular risk factors like hyperinsulinism and dyslipidemias particularly in a person with a family history of NIDDM or hypertension, deserves attention before overt diseases develop. [table: see text] PMID- 8249694 TI - A paradigm to link clinical research to clinical practice: the challenge in non insulin dependent diabetes mellitus. PMID- 8249695 TI - The beta-cell sulfonylurea receptor. PMID- 8249696 TI - Regulation of glucose transporters and the Na/K-ATPase by insulin in skeletal muscle. PMID- 8249697 TI - Insulin receptor: aspects of its structure and function. PMID- 8249698 TI - The diabetogenes concept of NIDDM. PMID- 8249699 TI - Recombinant DNA technology: a clinical revolution in pediatrics. PMID- 8249700 TI - Bone marrow transplantation. AB - BMT has been successfully utilized in clinical medicine for nearly 25 years. Successful accomplishment of BMT has severely taxed the research and clinical capabilities of medical scientists, but the attainment of a cure for fatal disease has amply justified the efforts expended. The major problems still to be solved include better conditioning regimens, better control of GVHD, and enlargement of donor pools. The control of problems created by this therapy demands a thorough understanding of the immune system. As this knowledge increases, useful manipulation of immunity to control the hundreds of other diseases caused by aberrant immune processes should become possible. Soon, genetically engineered marrow transplants should be possible. PMID- 8249701 TI - The current status of childhood lead toxicity. PMID- 8249702 TI - The follow-up of infants exposed to chloride-deficient formulas. PMID- 8249703 TI - Body composition. PMID- 8249704 TI - Bacterial meningitis in children: current concepts of neurologic management. PMID- 8249705 TI - Rett syndrome. PMID- 8249706 TI - Transcatheter closure of cardiac defects. PMID- 8249707 TI - Diuretic treatment agents. AB - Diuretics are important drugs in the treatment of fluid overload and other related conditions in infants and children. Their rational use depends on adequate knowledge of how they work and how they interact with other drugs and the changing physiology caused by normal growth and development and by various disease states. It is in the best interest of the patient for the physician prescribing these drugs to have appropriate knowledge of their mode of action. It is clear that with the exception of a few drugs the amount of appropriate data available on the use of these drugs in children is limited at best. As newer drugs become available, the physician caring for such children must be sure that there is some clear indication for its use in preference to those with which we have more experience. PMID- 8249708 TI - Nephrotic syndrome in childhood. AB - MLNS is the most common cause of nephrotic syndrome in childhood and has a very good prognosis. Other histologic types have a varying prognosis. Corticosteroid treatment induces a remission in the majority of patients with nephrotic syndrome. Every effort should be made to reduce the total dose of corticosteroid so as to minimize the potential for corticosteroid-induced side effects. PMID- 8249709 TI - The kidney in systemic disease: Part II--Autoimmune and vascular disorders. AB - The long-term outlook for the pediatric patient with lupus nephritis has improved dramatically over the past several decades. In 1968, Meislin and Rothfield compared childhood with adult SLE and found the prognosis to be worse when disease onset was at the age of 15 years or younger. Children with renal disease had only a 42% 5-year survival rate vs. an 82% survival rate in adults. Caeiro et al. found the 10-year survival rate in childhood to be only 48%. However, more recent data have demonstrated comparable results for patients of all ages. Ten year patient survival rates have improved to 69% to 85% as a result of better immunosuppressive therapy, more effective antibiotics, and improved control of hypertension. Despite this overall improvement, severe renal disease continues to complicate the course of 20% to 35% of patients with SLE originating in childhood and demands further investigation of newer therapeutic measures. PMID- 8249710 TI - Childhood immunization update. PMID- 8249711 TI - Renal stones and hypercalciuria. PMID- 8249712 TI - Anemia of prematurity: the prospects for avoiding blood transfusions by treatment with recombinant human erythropoietin. PMID- 8249713 TI - Advances in pediatric pharmacology and toxicology. PMID- 8249714 TI - Maternal cocaine use: impact on child health and development. PMID- 8249715 TI - The upper gastrointestinal series at a crossroads. AB - Radiologic examination of the upper gastrointestinal tract, or the upper gastrointestinal series, is at a crossroads. As we approach the 21st century, it is possible to envision two very different scenarios for the future of this examination. The gradual but steady decline in the number of upper gastrointestinal studies done during the past decade could continue or even accelerate until this technique is no longer used in diagnostic radiology. Or, spurred by new concerns about cost and safety in a rapidly changing political and economic environment, the upper gastrointestinal series could undergo a dramatic resurgence, supplanting endoscopy as the primary diagnostic test in patients with symptoms in the upper gastrointestinal tract. Radiologists may ultimately have a pivotal role in determining the fate of the upper gastrointestinal series by their commitment or lack of commitment to perpetuating this form of examination. In this article, we consider the status of the upper gastrointestinal series in current medical practice and the challenges we face if it is to remain a useful diagnostic technique in the 21st century. PMID- 8249716 TI - Coronary artery calcification: clinical significance and current methods of detection. AB - Coronary artery disease affects 1,500,000 Americans each year; 500,000 of these will die. The earliest detectable lesion of coronary atherosclerosis is the fatty streak. Later, crescent-shaped lipid plaques occur, which may rupture and produce either progressive stenosis or sudden occlusion with myocardial infarction. Calcium is deposited early in the formation of the atherosclerotic plaque, and calcification can be used as a marker of the atherosclerotic process. Many imaging techniques can be used to detect calcification of coronary arteries. The most promising are fluoroscopy, ultrafast CT, and intravascular sonography. Detection of calcification is most valuable in persons less than 40 years old in whom modification of risk factors may be important. In addition, the progression and possible regression of calcification can be used as an indicator of the atherosclerotic process. The absence of calcification in coronary arteries may diminish the need for further testing. PMID- 8249717 TI - Fast scanning and fat-suppression MR imaging of musculoskeletal disorders. AB - Two of the most exciting areas of development in MR imaging in the past several years have been rapid imaging and fat-suppression techniques. This article reviews the most widely available techniques for performing rapid imaging and fat suppression and summarizes current clinical applications in musculoskeletal imaging. PMID- 8249718 TI - Pulmonary eosinophilic granuloma. PMID- 8249719 TI - Serial CT in fibrosing alveolitis: prognostic significance of the initial pattern. AB - OBJECTIVE: In patients with fibrosing alveolitis, a reticular pattern on CT scans of the lungs correlates with histologic findings of fibrosis, whereas a ground glass pattern has been reported to correlate with reversible inflammatory disease. The majority of patients with fibrosing alveolitis have a ground-glass component visible on CT scans, but response to therapy is infrequent. The aim of this study was to evaluate the prognostic significance of the relative extents of ground-glass and reticular patterns by analyzing serial changes in these CT appearances. SUBJECTS AND METHODS: Serial CT scans were analyzed in 56 patients (21 with idiopathic pulmonary fibrosis and 35 with fibrosing alveolitis associated with systemic sclerosis). The relative extents of ground-glass and reticular patterns were assessed on the initial CT scan. Overall extent of abnormal lung and distribution of disease on initial CT scans were also categorized. Changes in extent and appearance of disease were evaluated in paired anatomically comparable CT sections and assessed independently by two observers; the median interval between scans was 16 months. Serial changes on CT were examined in relation to the initial pattern, extent, and distribution of disease seen on CT scans and in relation to trends in results of concurrent pulmonary function tests. RESULTS: Changes in the extent of disease were due to regression of a ground-glass pattern in 18 patients, an increase in a reticular pattern in nine patients, and an increase in a ground-glass pattern in five patients. A reticular pattern did not regress in any patient. In treated patients, diminution in extent of disease, shown as regression of a ground-glass pattern, was seen most frequently when a ground-glass pattern was the most extensive abnormality at initial scanning (p < .002), independent of extent or distribution of disease seen on CT scans and the type of fibrosing alveolitis. When a ground-glass pattern was associated with an equally extensive reticular pattern, the extent of disease diminished with therapy in a minority of patients (5/13). Improvement in results of pulmonary function tests was associated with regression of a ground glass pattern in the majority of patients (8/11). CONCLUSION: These findings indicate that the prognostic significance of a ground-glass pattern depends on the extent of an associated reticular pattern and is independent of the extent and distribution of disease. PMID- 8249720 TI - Previous mammograms in patients with impalpable breast carcinoma: retrospective vs blinded interpretation. 1993 ARRS President's Award. AB - OBJECTIVE: We examined differences between blinded and retrospective reviews of screening mammograms obtained before a mammogram that resulted in the diagnosis of an impalpable breast carcinoma. MATERIALS AND METHODS: We reviewed 152 previous mammograms in 73 patients in whom impalpable breast carcinomas were subsequently detected on later mammograms. The earlier studies were interpreted in two ways: (1) blindly (without knowledge that carcinoma was subsequently detected) and (2) retrospectively (with the mammogram showing the carcinoma for comparison). The two interpretations were then compared with regard to the presence of carcinoma, recommendations for biopsy, parenchymal density, histologic characteristics of the tumor, lymph node status, and film quality. RESULTS: When we did a blinded review of the mammograms obtained before the diagnostic mammograms, the previous study was interpreted as showing evidence of carcinoma in 30 patients (41%). For the remaining 43 patients (59%), the findings of the most-recent previous mammogram were interpreted as normal or benign by the blinded reviewers; however, the retrospective reviewers thought evidence of cancer was visible in 25 of these patients (34%). Differences between blinded and retrospective interpretations were statistically significant. In patients in whom evidence of tumor was thought to be present on retrospective review but not on blinded review, the majority of mammographic abnormalities were asymmetric densities on the most-recent previous examination. This was true whether or not the retrospective reviewers thought that the mammographic finding warranted earlier biopsy. The histologic characteristics and lymph node status among patients in whom mammograms were interpreted retrospectively as showing evidence of tumor were no different from those among patients with no evidence of tumor. CONCLUSION: Our results show that impalpable breast carcinomas are frequently evident in retrospect on previous mammograms. However, because many are manifested only as an asymmetric density, these may not necessarily be true radiologic errors. Failure to detect a retrospectively visible abnormality on a screening mammogram is not necessarily negligent, and retrospective reviews do not reflect the everyday practice of screening mammography. PMID- 8249721 TI - Tubular carcinoma of the breast: mode of presentation, mammographic appearance, and frequency of nodal metastases. AB - OBJECTIVE: Tubular carcinoma of the breast is a distinct, well-differentiated histologic subtype of infiltrative adenocarcinoma. The purpose of this study was to determine the typical mode of presentation, mammographic appearance, and frequency of metastases to the axillary lymph nodes. MATERIALS AND METHODS: We retrospectively analyzed the clinical records, mammograms, and histologic slides of 20 cases of proved tubular carcinoma of the breast in 20 women. These patients were identified by computerized search of our pathology data base from 1984 to 1993. Histologic findings were reviewed in all cases and correlated with the mammographic findings. RESULTS: Thirteen (65%) of the 20 women had impalpable tubular cancers discovered on screening mammograms. Seven patients (35%) had mammography because of a palpable tumor. Abnormalities were seen on mammograms in 16 patients (80%). These consisted of a mass alone in 13 (65%), a mass with microcalcifications in two (10%), and calcifications associated with architectural distortion and asymmetric density in one (5%). Most masses (11/15, 73%) were irregular or spherical and had spiculated margins. The size of the masses ranged from 3 to 19 mm (median, 8 mm). Metastatic carcinoma was found in the axillary lymph nodes of four (29%) of 14 patients who had axillary lymph node dissections, three of whom had a primary tumor 1 cm or less in diameter. CONCLUSION: Tubular cancers were most frequently detected as impalpable abnormalities seen on mammograms, emphasizing the importance of screening mammography. However, tubular carcinomas did not have a unique appearance on mammograms that would allow differentiation from other carcinomas. Lymph node metastases were more common than has been previously reported for this type of tumor, indicating that axillary lymph node dissections should continue to be performed. PMID- 8249722 TI - Mammographic changes in postmenopausal women undergoing hormonal replacement therapy. AB - Increasing numbers of postmenopausal women are undergoing hormonal replacement therapy. In this pictorial essay, we present the spectrum of mammographic changes seen in these women. These changes include symmetric and asymmetric increase in breast density, increase in size of fibroadenomas, and development or increase in size of cysts. Our examples illustrate that differentiation of hormonal therapy changes from neoplasm can occasionally be problematic when a focal density or mass is seen on mammograms. Furthermore, reexamination of published reports and our cases suggests that treatment with estrogen alone promotes enlargement of cysts and fibroadenomas, whereas treatment with a combination of estrogen and progesterone is more likely to be associated with diffuse increase in density. PMID- 8249723 TI - Spiral CT of the abdomen: artifacts and potential pitfalls. AB - Spiral CT has been advocated as a major advancement in CT technology. Spiral CT scanning is rapid, the volume of IV contrast material can potentially be decreased, and high-quality coronal, sagittal, and three-dimensional reconstruction is possible. However, when compared with dynamic CT, because of the shorter acquisition time of spiral CT, vascular and organ enhancement on spiral CT scans is more dependent on factors that affect the delivery of contrast material into the bloodstream and on the time that scanning begins after the start of injection of contrast material. Additionally, organs other than the liver are scanned earlier than they typically are during a dynamic study, particularly the kidneys and spleen. In our experience to date, these aspects of spiral scanning have led to several difficulties in the interpretation of spiral CT scans. This essay illustrates potential pitfalls in the interpretation of spiral CT scans associated with scanning too soon after injection of contrast material. PMID- 8249724 TI - MR imaging with i.v. superparamagnetic iron oxide: efficacy in the detection of focal hepatic lesions. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of superparmagnetic iron oxide (SPIO) in the detection of focal hepatic lesions on MR images. SUBJECTS AND METHODS: The study included 21 patients with 115 focal hepatic lesions and eight patients without focal hepatic lesions. T1- and T2 weighted MR images were obtained at 1.5 T before and 60 min after the end of injection of an SPIO agent. Contrast-enhanced CT scans were obtained in all patients within 10 days after MR imaging. The effect of SPIO on the signal intensity of the liver and spleen was assessed by using quantitative analysis of the region of interest. Efficacy was evaluated by using multiple criteria and unenhanced and SPIO-enhanced images. Evaluations included subjective assessment of image quality, counting the number of lesions detected, and statistical analysis of quantitative changes in the signal intensity of lesions and of normal liver. RESULTS: By all criteria, SPIO-enhanced T2-weighted MR images were superior to unenhanced T2-weighted images and to contrast-enhanced CT scans. Conversely, by all criteria, SPIO-enhanced T1-weighted MR images were worse than unenhanced T1-weighted images and contrast-enhanced CT scans. The mean lesion-to liver contrast on T2-weighted images was 317% on unenhanced images and 1745% on SPIO-enhanced images. For T1-weighted, the mean contrast was 26% on unenhanced images and 18% on SPIO-enhanced images. CONCLUSION: SPIO is an efficacious contrast agent for the detection of focal hepatic lesions when T2-weighted MR images are used. PMID- 8249725 TI - Diagnosis of appendicitis: imaging findings in patients with atypical clinical features. AB - Acute appendicitis is usually diagnosed on the basis of signs, symptoms, and results of simple laboratory tests. However, in patients with atypical clinical features, imaging studies play a significant role in preoperative diagnosis and determination of proper treatment. In this pictorial essay, we present a spectrum of imaging findings in patients whose clinical features were unusual because of the abnormal position of the appendix, the presence of other illnesses, or the presence of complications in children. Appendicitis was not initially considered on clinical presentation in any of our cases. In most cases, the correct diagnosis of appendicitis was made or, at least in retrospect, could have been made on the basis of imaging findings. PMID- 8249726 TI - Pharyngography after head and neck irradiation: differentiation of postirradiation edema from recurrent tumor. AB - OBJECTIVE: In patients previously irradiated for head and neck carcinomas, persistent soft-tissue thickening in the larynx and pharynx makes it difficult to distinguish between postirradiation edema and recurrent epidermoid carcinoma. The goal of this work was to characterize the standard double-contrast pharyngographic appearance after irradiation and to differentiate this appearance from that of recurrent or residual neoplasms. MATERIALS AND METHODS: The posttreatment pharynogograms in 43 patients treated with radiotherapy for malignant tumors of the head and neck were retrospectively reviewed. The pharynx had been included in the radiation field in each case. We characterized the posttherapy appearance and identified radiographic signs that could be used to distinguish normal postirradiation edema from recurrent tumor. RESULTS: Soft tissue thickening of normal laryngeal and pharyngeal structures and laryngotracheal aspiration were common nonspecific findings on posttherapy pharyngograms, each occurring in 39 of 43 patients. Epiglottic enlargement and laryngotracheal aspiration did not correlate with the presence of residual or recurrent neoplasm. Asymmetric swelling was more common ipsilateral to the original neoplasm and did not necessarily indicate a malignant tumor. Mucosal irregularity, ulceration, and a demonstrable focal mass were the only significant predictors of recurrent or residual tumor; at least one of the three findings was present in 17 of 19 patients with neoplasm correctly identified by pharyngography. CONCLUSION: Double-contrast pharyngography is a useful means of diagnosing pharyngeal tumors in patients who have had radiation therapy. Radiographic signs of neoplasm (ulcer, mucosal irregularity, and a focal mass) can be used to distinguish recurrent tumor from nonmalignant changes after irradiation. PMID- 8249727 TI - CT imaging of abdominal hernias. AB - Most abdominal hernias can be diagnosed on the basis of findings on physical examination or plain films and barium studies. However, diagnostic dilemmas can arise when patients are obese or have had surgery. Cross-sectional CT scans can show hernias and the contents of the peritoneal sac. More important, CT findings can be used to diagnose unsuspected hernias and to distinguish hernias from masses of the abdominal wall, such as tumors, hematomas, abscesses, undescended testes, and aneurysms. This essay illustrates the CT findings in diaphragmatic hernias, internal hernias, and abdominal wall hernias. PMID- 8249728 TI - Acquired C1 esterase inhibitor deficiency causing intestinal angioedema: CT appearance. PMID- 8249729 TI - Bone marrow reconversion in adults who are smokers: MR Imaging findings. AB - OBJECTIVE: Conversion of bone marrow in the extremities from red to yellow is a normal maturation process. Marrow reconversion is present when expected yellow marrow is replaced with active red marrow, and it tends to occur at times of physiologic stress. The significance of this finding on MR imaging is not always clear. Accordingly, we studied the prevalence of bone marrow reconversion in adults undergoing MR imaging of the knee for reasons other than marrow disorders. Possible relationships between marrow reconversion and age, sex, weight, and smoking were sought. MATERIALS AND METHODS: Fifty-nine outpatients over 24 years of age who had no evidence of hematopoietic disorders and for whom MR images of the knee were available for review were selected for study. Patients younger than 25 years old were eliminated to ensure that all patients in the study were old enough to have achieved normal adult marrow conversion. The 59 subjects were classified according to their smoking histories. Twenty-eight were nonsmokers, 10 were intermediate smokers, and 21 were heavy smokers. The MR images were classified as showing either red marrow (reconverted from yellow to red) or yellow marrow (converted) by two independent observers who had no knowledge of the patients' smoking histories. RESULTS: MR images of six heavy smokers, two intermediate smokers, and one nonsmoker showed evidence of reconversion. A statistically significant association between marrow reconversion and heavy smoking (p = .02) was found. Reconversion also was found to be more prevalent in patients less than 39 years old and in obese women who smoked. No association was found between weight, sex, or obese male smokers and reconversion. CONCLUSION: Our results show marrow reconversion at the knee is most prevalent in heavy smokers, younger patients, and especially obese women who smoke heavily. In these patients, marrow reconversion can be a normal finding on MR imaging. In other patients, other causes should be considered. PMID- 8249730 TI - CT of the spine: value of reformatted images. AB - Recent improvements in CT software allow rapid and high-quality two-dimensional reformatting of scans of the spine. Reformatting involves computer interpolation of two-dimensional multislice images acquired during routine axial CT scanning in order to generate a three-dimensional volume of data. From this block of data, new images are created and displayed in any desired plane. Reformatted images are acquired within minutes following axial CT scanning. PMID- 8249731 TI - Glenoid labral tears: prospective evaluation with MRI imaging, MR arthrography, and CT arthrography. AB - OBJECTIVE: We prospectively compared MR imaging, MR arthrography, and CT arthrography to determine the sensitivity of each technique in detecting glenoid labral tears and in determining whether the labrum is detached or degenerated. SUBJECTS AND METHODS: Thirty patients 19-39 years old (mean, 27 years old) who had either signs and symptoms of shoulder instability or shoulder pain of unexplained origin were referred for diagnostic imaging. Each patient underwent MR imaging, followed by MR arthrography after intraarticular injection of 25 ml of a dilute solution of gadopentetate dimeglumine. Twenty-eight of thirty patients underwent CT arthrography after intraarticular injection of air and radiographic contrast material. Each patient also underwent arthroscopy or open surgery. RESULTS: At surgery, labral tears were found in 28 patients; a detached fragment was found in 26 patients. The labrum was found to be degenerated in 18. A labral tear was detected on MR images in 26 (93%) of 28, on MR arthrograms in 27 (96%) of 28, and on CT arthrograms in 19 (73%) of 26. A detached labral fragment was detected on MR images in 12 (46%) of 26, on MR arthrograms in 25 (96%) of 26, and on CT arthrograms in 13 (52%) of 25. Labral degeneration was detected on MR images in two (11%) of 18, on MR arthrograms in 10 (56%) of 18, and on CT arthrograms in four (24%) of 17. MR arthrography was the best of the three imaging techniques for showing the inferior part of the glenoid labrum and inferior glenohumeral ligament. CONCLUSION: MR arthrography and MR imaging both showed labral tears with greater sensitivity than CT arthrography did. MR arthrography was the most sensitive of the three techniques for detecting a detached labral fragment and labral degeneration. Furthermore, MR arthrography afforded the best visualization of the inferior part of the labrum and the inferior glenohumeral ligament. MR imaging and MR arthrography also enabled direct visualization of rotator cuff disease and other unsuspected associated abnormalities. PMID- 8249732 TI - Tear of the posterior tibial tendon causing asymmetric flatfoot: radiologic findings. AB - OBJECTIVE: The purpose of this study was to describe the preoperative radiographic appearance of the acquired asymmetric flatfoot caused by a tear of the posterior tibial tendon. These radiographic changes reflect the loss of tendon function and the development of flatfoot deformity. SUBJECTS AND METHODS: Preoperative radiographs of 30 patients (mean age, 48 years) with surgically proved complete tears of the posterior tibial tendon were evaluated. Erect anteroposterior and lateral views of the feet were obtained in all patients and were frequently supplemented with anteroposterior views of the ankles to evaluate valgus tilt. Calcaneal plantar, talocalcaneal, and talometatarsal angles were measured on radiographs and compared with the measurements in normal control subjects. Additional osseous and soft-tissue abnormalities were also evaluated. RESULTS: Findings on preoperative radiographs of the foot were abnormal in 50% of patients. These abnormalities included decreased calcaneal plantar angle (50%), increased lateral talometatarsal angle (47%), increased anterior talocalcaneal angle (43%), and increased lateral talocalcaneal angle (13%). Bone and soft tissue abnormalities included osteoporosis (37%), medial soft-tissue swelling (27%), tarsal osteoarthritis (20%), distal tibial proliferative changes (7%), and the presence of accessory navicular bones (17%). CONCLUSION: Multiple abnormalities of the intertarsal relationships are seen as a result of tears of the posterior tibial tendon with the development of hindfoot valgus, midfoot abduction, and forefoot pronation. PMID- 8249733 TI - Spring ligament of the ankle: normal MR anatomy. AB - The plantar calcaneonavicular or spring ligament is visualized inconsistently and incompletely on routine MR images of the foot. This ligament is a vital stabilizer of the longitudinal arch of the foot, providing support for the head of the talus, which rests on the ligament's central portion. Laxity or rupture of the spring ligament permits plantar flexion of the talus. This motion results in valgus alignment of the calcaneus and a flatfoot deformity (pes planovalgus). Laxity or rupture of the spring ligament can develop in cases of chronic dysfunction of the posterior tibial tendon. In rupture of the posterior tibial tendon, surgical management may include plication of the spring ligament in addition to repair or reconstruction of the tendon to stabilize the medial column of the foot. Thus, the status of the spring ligament can be a significant consideration in preoperative planning. This pictorial essay illustrates the normal MR anatomy of the spring ligament, the planes of imaging required for optimal depiction of the ligament, and the neighboring structures with which the ligament can be confused. PMID- 8249734 TI - Gastric duplication with hemoptysis: CT findings. PMID- 8249735 TI - Prenatal sonographic diagnosis of scalp edema/cephalohematoma mimicking an encephalocele. PMID- 8249736 TI - The pelvis after cesarean section and vaginal delivery: normal MR findings. AB - OBJECTIVE: In postpartum women, pain and fever can indicate serious complications, such as phlegmon or abscess of the parametrium or bladder flap, ovarian vein thrombosis, or uterine dehiscence. The purpose of this study was to ascertain the MR appearance of the normal pelvis after uncomplicated cesarean section and vaginal delivery. SUBJECTS AND METHODS: Twenty-two patients (14 who had cesarean sections and eight who had vaginal deliveries) were imaged with a superconducting 0.35-T MR system 1-5 days after delivery. Sagittal and axial T1 weighted, proton density-weighted, and T2-weighted spin-echo images were acquired. Phase reconstruction of the axial T1-weighted sagittal images was performed to confirm flow in the ovarian veins. The ovarian veins, parametrium, bladder flap, endometrium, and uterine incision were analyzed on all images. RESULTS: The uterine incision was well seen in all 14 patients who had cesarean section. The signal characteristics of the incision suggested subacute hematoma within the myometrium, and no myometrial defects were seen. In all 13 patients with a low transverse incision, a hematoma of the bladder flap was seen. Patency was confirmed in the majority of ovarian veins (39/44). No evidence of parametrial edema or mass, which can be seen in abscess or phlegmon, was found in any of the 22 patients. The ovarian perivenous fat did not show edema, which can indicate acute ovarian vein thrombosis. Increased signal in the endometrial cavity was seen in all patients; the signal characteristics were similar to those of subacute hematoma, consistent with clinical findings. CONCLUSION: Normal findings of the postpartum pelvis on MR imaging are subacute hematoma in the uterine incision, bladder flap, and endometrial cavity. Patent ovarian veins are demonstrated in most (89%) cases. PMID- 8249737 TI - Sonography of ocular choroidal melanomas. AB - Ocular choroidal melanomas lend themselves particularly well to evaluation by sonography. This essay illustrates the different appearances of ocular melanoma as depicted with sonographic imaging, including conventional gray-scale sonography and duplex Doppler, color flow Doppler, and three-dimensional display. PMID- 8249738 TI - Toluene abuse causing reduced MR signal intensity in the brain. PMID- 8249739 TI - MR imaging in a patient with Meniere's disease. PMID- 8249740 TI - Parathyroid scintigraphy with iodine-123 and 99mTc-sestamibi: imaging findings. AB - Subtraction scintigraphy with 99mTc-pertechnetate and 201Tl-chloride is widely used for localizing the parathyroid gland, but the sensitivity of this method averages only 55%. The sensitivity of scintigraphy is improved by using a combination of 99mTc-sestamibi and iodine-123. Parathyroid adenomas can also be localized with 99mTc-sestamibi alone by subtracting early images from late sequential images. The fast washout of 99mTc-sestamibi from the thyroid gland allows visualization of the parathyroid gland. The purpose of this essay is to illustrate the findings of abnormalities of the parathyroid glands when using a combination of 99mTc-sestamibi and iodine-123. PMID- 8249741 TI - Cervical carotid MR angiography with multiple overlapping thin-slab acquisition: comparison with conventional angiography. AB - OBJECTIVE: Multiple overlapping thin-slab acquisition (MOTSA) is a hybrid time-of flight MR angiography technique that combines the advantages of two- and three dimensional methods. A venetian blind-like artifact, resulting from variations in signal intensity at slab boundaries, is a potentially significant disadvantage. The objective of this study was to determine the accuracy of MOTSA MR angiography for the measurement of stenosis and to evaluate the effectiveness of modifications to reduce the venetian blind artifact. SUBJECTS AND METHODS: Fifty one consecutive patients undergoing conventional and MR angiography of the carotid arteries for suspected atherosclerotic disease were studied. Methods established in the North American Symptomatic Carotid Endarterectomy Trial (NASCET) were used to measure stenosis. Measurements from the conventional angiograms were compared with those from the MR angiograms. Modifications of the MR angiographic technique that were instituted after the initial 26 patients to reduce the venetian blind artifact included new acquisition parameters and the use of a postprocessing algorithm. RESULTS: After modification, highly significant improvements in Spearman ranked correlation coefficients were observed for MR angiography interobserver comparison (.573 to .974, p < .001) and comparison of conventional and MR angiography (.774 to .949, p < .001). A corresponding reduction was observed in the standard deviation of the difference measurement for percent stenosis for both interobserver (34.5% to 11.7%, p < .001) and intertechnique (22.6% to 12.6%, p < .001) comparisons. In the postmodification group, the interobserver correlation coefficients for conventional and MR angiography were not significantly different (.969 vs .974). The areas under the receiver-operating-characteristic curves used to predict surgically significant disease for the premodification and postmodification groups were 0.79 and 0.98, respectively, for observer A and 0.85 and 0.95, respectively, for observer B. CONCLUSION: Improved acquisition parameters and postprocessing significantly reduce the severity of the venetian blind artifact and increase the accuracy of MOTSA MR angiography in the measurement of percent stenosis. With these improvements, MOTSA MR angiography shows promise for noninvasive evaluation of stenosis of the carotid arteries. PMID- 8249742 TI - Correlation of arterial and venous blood flow in the mesenteric system based on MR findings. 1993 ARRS Executive Council Award. AB - OBJECTIVE: The purpose of this study was to determine if a consistent relationship exists between (1) flow in the portal vein and superior mesenteric vein and (2) flow in the mesenteric arteries. SUBJECTS AND METHODS: Cine phase contrast methods were used to determine volumetric flow in the celiac axis, superior mesenteric artery, portal vein, and superior mesenteric vein in 11 healthy volunteers. RESULTS: The mean (+/- SD) volumetric flow rates in the celiac axis, superior mesenteric artery, superior mesenteric vein, and portal vein were 9.9 +/- 3.2, 6.0 +/- 1.8, 5.7 +/- 2.0, and 13.7 +/- 1.8 ml/min per kilogram, respectively. A strong correlation was found between measured mesenteric arterial blood flow, defined as the sum of flow in the celiac axis and the superior mesenteric artery, and portal venous flow (r = .97, p = .00002). Good agreement was found between flow in the superior mesenteric artery and that in the superior mesenteric vein (r = .98; p = .00001; bias = 20 ml/min). CONCLUSION: There is a consistent relationship between MR measurements of flow in the portal or superior mesenteric vein and the measured flow in the arteries supplying those veins. Flow in the superior mesenteric vein is an accurate predictor of flow in the superior mesenteric artery. PMID- 8249743 TI - Protein S deficiency: imaging findings. AB - Protein S deficiency is an autosomal-dominant inherited disorder of coagulation associated with spontaneous and recurrent venous thrombosis. Approximately 5% of patients with deep venous thrombosis of the lower extremities who are less than 45 years old have a deficiency of protein S. Patients frequently have spontaneous and recurrent deep venous thrombosis and pulmonary embolism during young adulthood. Thrombosis occurs less commonly in the splanchnic and cerebral veins. Arterial thrombosis is rare but is seen in the cerebral circulation. Radiologists should include protein S deficiency in the differential diagnosis of unexplained thrombosis in young patients. This pictorial essay illustrates the range of imaging findings encountered in patients with this disorder. PMID- 8249744 TI - Is bilateral lower extremity compression sonography useful and cost-effective in the evaluation of suspected pulmonary embolism? AB - OBJECTIVE: Because deep venous thrombosis is clinically linked with pulmonary embolism and often treated similarly, we sought to assess the usefulness of obtaining bilateral lower extremity compression sonograms when findings on ventilation-perfusion lung scans indicate a low or indeterminate probability of pulmonary embolism. Demonstration of deep venous thrombosis would provide a rationale for treating both pulmonary embolism and deep venous thrombosis. MATERIALS AND METHODS: Two hundred twenty-three consecutive patients with suspected pulmonary embolism had ventilation-perfusion lung scans and concurrent bilateral lower extremity compression sonograms; 34 also had pulmonary arteriography. RESULTS: In 75 cases, the results of ventilation-perfusion lung scanning indicated an indeterminate probability of pulmonary embolism. Evidence of thrombosis was seen on sonograms in 11 of these 75. In the remaining 64, 17 underwent pulmonary arteriography and four (24%) had pulmonary embolism. Findings on lung scans indicated a low probability of pulmonary embolism in 70 of 223 patients. Evidence of thrombosis was seen on sonograms in 11 of these 70. Five of the remaining 59 underwent pulmonary arteriography and one (20%) had pulmonary embolism. According to the 1993 Medicare Fee Schedule, if all 145 patients whose lung scans were nondiagnostic had sonography and only those with normal sonograms had pulmonary arteriography, the professional and hospital charges would be $359,552. If all 145 had pulmonary arteriography without sonography, the charges would be $395,031. CONCLUSION: If ventilation-perfusion lung scans indicate a low or an indeterminate probability of pulmonary embolism and bilateral lower extremity compression sonography is performed, only those patients with normal sonographic findings would need further study. Thus, 15% (22/145) of patients could be spared pulmonary arteriography, and the estimated savings in cost would be 9%. PMID- 8249745 TI - Blinded comparison of biopsy needles and automated devices in vitro: 1. Biopsy of diffuse hepatic disease. AB - OBJECTIVE: The purpose of this study was to compare several of the commonly used needles with several of the new automated biopsy devices (biopsy guns) for biopsy of diffuse hepatic disease. MATERIALS AND METHODS: Nine different biopsy needles or automated devices were each used to do three biopsies of 10 cadaveric livers. The specimens were reviewed in a blinded fashion by a pathologist who did not know which needle or device was used, and they were compared on the basis of a previously published histopathologic grading scale. RESULTS: The three conventional biopsy needles (16-gauge Jamshidi, 18-gauge Sure-Cut, and 14-gauge Tru-Cut) obtained a large amount of tissue with an average of 4.1 intact portal triads per biopsy. The 18-gauge Biopty gun obtained equivalent results. The 18 gauge Autovac gun with a 2-cm biopsy depth did not obtain any tissue in 18.5% of attempts. The 14- and 16-gauge Biopty guns and the 18-gauge Autovac gun with a 4 cm biopsy depth performed best with respect to fragment size and number of intact portal triads. CONCLUSION: Automated biopsy devices can provide more diagnostic specimens than can manual or conventional needles in biopsy for diffuse hepatic disease. PMID- 8249746 TI - Blinded comparison of biopsy needles and automated devices in vitro: 2. Biopsy of medical renal disease. AB - OBJECTIVE: The purpose of this study was to compare several of the commonly used manual biopsy needles with several of the new automated biopsy devices (biopsy guns) for biopsy of medical renal disease. MATERIALS AND METHODS: Ten different biopsy needles or automated devices were each used to do two biopsies of 10 cadaveric kidneys. The specimens were reviewed in a blinded fashion by a pathologist using a previously published histopathologic scale. RESULTS: Of the four conventional biopsy needles tested (16-gauge Jamshidi, 18-gauge Sure-Cut, 14 gauge Tru-Cut, and 14-gauge Vim Silverman), the Jamshidi needle obtained the greatest average number of glomeruli (5.64). Results similar to those obtained with the conventional needles were obtained with the 16- and 18-gauge Biopty and Ultra-Cut biopsy guns. The 18-gauge Autovac gun with either a 2- or 4-cm depth of biopsy suffered from a significant number of biopsies from which no tissue was obtained. The 14-gauge Biopty gun was clearly superior, leading in all graded categories including the average number of glomeruli (8.11) per biopsy. CONCLUSION: The automated biopsy device, or biopsy gun, can provide more diagnostic specimens than can manual or conventional needles in biopsy for medical renal disease. PMID- 8249747 TI - Diagnosis of renal cell carcinoma: value of fine-needle aspiration cytology in patients with metastases or contraindications to nephrectomy. AB - OBJECTIVE: A retrospective study of imaging-directed percutaneous fine-needle aspiration cytology of focal renal lesions was performed. The objectives were to determine the efficacy and safety of renal aspiration biopsy in establishing the diagnosis of renal cell carcinoma and avoiding nephrectomy in patients with a renal mass and disseminated metastases or with a renal mass and relative contraindications to nephrectomy. MATERIALS AND METHODS: Between September 1987 and September 1991, 55 consecutive patients had 57 imaging-directed renal aspiration biopsies at our institution. We examined the medical records, pathology reports, imaging studies, and follow-up information of the 23 patients who had the procedure because of disseminated metastases (10 patients) or relative contraindications to nephrectomy (13 patients). Preliminary diagnoses were based on the cytopathology, and final diagnoses were based on the surgical pathology (six patients) or clinical course over more than 6 months (17 patients). RESULTS: Cytopathologic findings were true-positive in 12 patients, true-negative in eight, false-negative in three, and false-positive in none. The sensitivity was 80%, the specificity was 100%, and the accuracy was 87%. Of the 10 patients with a renal mass and disseminated metastases, aspiration cytology failed to show malignancy in only one. None of these patients had surgery, and all died within 1 year. Of the 13 patients with relative contraindications to surgery, seven were treated without surgery. The cytologic diagnosis was renal cell carcinoma in two of the seven, and these two patients have since died. Cytopathology was negative for renal cell carcinoma in five of the seven, and subsequent imaging studies in these patients have shown no renal mass enlargement or metastases 2-3 years after biopsy (three patients) or the patients have died of severe heart disease (two patients). Nephrectomy was performed in the remaining six patients, and no evidence of disease had been found 2-4 years after removal of two oncocytomas and four renal cell carcinomas. There were no significant complications. CONCLUSION: Percutaneous renal aspiration biopsy cytology is accurate, safe, and useful in establishing the diagnosis of renal cell carcinoma in patients with disseminated metastases or relative contraindications to surgery. PMID- 8249748 TI - Adjustable-depth biopsy device: comparison with a single-depth biopsy gun in cadaveric organs. PMID- 8249749 TI - Authentication and management of radiologic reports: value of a computer workstation integrated with a radiology information system. AB - An increasing number of radiology departments are using computers to facilitate management of radiologic information. Transcription, storage, and printing of radiologic reports are among the primary functions of a radiology information system. Consequently, manual signature of radiologic reports is being replaced by on-line electronic authentication. However, the utilities provided on most radiology information systems to review, edit, and otherwise manipulate radiologic reports are relatively crude compared with commercial word-processing and data-base management software available for personal computers. We have developed a personal computer software system that is integrated with our existing radiology information system; expedites the radiologist's task of reviewing, editing, and authenticating (i.e., signing) radiologic reports; provides a teaching file data base on the workstation into which radiologic reports and patients' demographics can be instantly transferred; and provides a similar data base to facilitate follow-up on those examinations deemed appropriate for quality-assurance procedures. These features improve the radiologist's efficiency and increase his or her willingness to more fully exploit the intended purposes of a radiology information system. PMID- 8249750 TI - Data interface between a radiology information system and a computed radiography system using a personal computer and standard software. AB - We describe the implementation of a simple data interface between a radiology information system and a computed radiography system that uses personal computers and standard software. The radiology information system was developed in house and runs in a local area network of personal computers. The computed radiography system is connected to a picture archiving and communication system. We have implemented a software data interface on a microcomputer, allowing automated transfer of patient data from the radiology information system program to the computed radiography scheduling program. The interface adds essential information used by the picture archiving and communication system to handle work lists, routing, and archiving algorithms. We have improved the user interface, shortened the scheduling time, enabled coherence of data bases, and eased the use of computed radiography and examination routing in the picture archiving and communication system. We have been using this interface for more than 1 year without difficulties. This custom solution addresses the problem of interconnecting existing equipment, avoiding proprietary restrictions or the lack of effective standards. This approach can be applied to any radiology environment that uses personal computers. PMID- 8249751 TI - Do citation conventions influence coauthorship patterns? AB - The main purpose of contributing to radiologic publications should be to advance the state of knowledge or increase awareness of issues useful in radiologic practice and research. According to experts on medical writing, however, the pride of discovery and the communication of scientific information for the enrichment of public knowledge are not the sole motives of authors. Success in the profession, prestige, and tenure have long been associated with prolific publishing in peer-reviewed journals. A proof of this association is the documented rise through recent years in the number of authors per paper. The phenomenon has been attributed in part to the increasing complexity and labor intensiveness of biomedical research, but this speculation remains unproved, and it is believed that often the allocation of authorship has a volitional component. PMID- 8249752 TI - History of diagnostic radiology at the Mayo Clinic. PMID- 8249753 TI - Intraabdominal metastatic thymoma. PMID- 8249754 TI - Primary liposarcoma of the liver. PMID- 8249755 TI - Paraumbilical to pelvic portosystemic collateral: diagnosis with color Doppler sonography. PMID- 8249756 TI - Unsuspected malignant pheochromocytoma: appearance on thallium and gallium scans. PMID- 8249757 TI - Extracranial cerebellum in Chiari II Malformation. PMID- 8249758 TI - MR imaging: a precision instrument for measuring the pituitary stalk? PMID- 8249759 TI - Deep venous thrombosis complicating sonographically guided compression repair of a pseudoaneurysm of the common femoral artery. PMID- 8249760 TI - Remote transgraft hemorrhage complicating thrombolysis with tissue plasminogen activator. PMID- 8249761 TI - The sticky biopsy specimen: a saline solution. PMID- 8249762 TI - The generic drug approval process. PMID- 8249763 TI - Blood lead screening: the argument for it. PMID- 8249764 TI - Blood lead screening: the argument against it. PMID- 8249765 TI - Home-mixed vs. commercial oral rehydration solutions. PMID- 8249766 TI - Dermal cylindromas. PMID- 8249767 TI - Oral corticosteroids and postherpetic neuralgia. PMID- 8249768 TI - Sacroiliac sprain. PMID- 8249769 TI - Anterior knee pain. PMID- 8249770 TI - Anterior knee pain. PMID- 8249771 TI - Use of body-mass index in elderly patients with bone loss. PMID- 8249772 TI - Diary from a week in practice. PMID- 8249773 TI - Microalbuminuria and diabetes mellitus. AB - Small concentrations of albumin detected in the urine predict renal dysfunction and reflect vascular abnormalities such as atherosclerosis, retinopathy and, probably, neuropathy. Although microalbuminuria is not specific for diabetic complications, it has been most extensively studied in diabetics. The rate of urinary albumin excretion can also be used to determine therapeutic response to pharmacologic and lifestyle interventions such as diet, smoking cessation and physical activity. The pathophysiology of microalbuminuria and its clinical significance in diabetes is presented, along with a discussion of measurement issues and implications for clinical management. An algorithm for the evaluation of diabetic patients is included. PMID- 8249774 TI - Assessment of the child with failure to thrive. AB - Failure to thrive is a presenting symptom, rather than a diagnosis. Organic failure to thrive may be caused by defects in food assimilation, excessive loss of ingested calories, increased energy requirements or prenatal insults. Nonorganic failure to thrive may occur accidentally, or it may be related to economic or emotional deprivation, as well as to child abuse or neglect. Assessment of a child with failure to thrive includes a comprehensive history and a thorough physical examination. Some basic laboratory screening tests may be helpful in the evaluation. More extensive investigation may be required, based on the clinical findings and the results of the initial screening tests. Early intervention is essential, because children may develop significant long-term sequelae from nutritional deficiency. PMID- 8249775 TI - Exposure to hazardous substances and reproductive health. Agency for Toxic Substances and Disease Registry. AB - The extent to which environmental or occupational chemical exposures affect reproductive health remains largely unknown. Teratogenic effects usually take place during critical periods of organogenesis. Common concerns of patients include the possible effects of drinking contaminated water on the fetus as well as the breast-fed infant, and the possible effects of a chemical exposure on fertility and pregnancy. PMID- 8249776 TI - Recurrent spontaneous abortion: evaluation and management. AB - Recurrent, or habitual, spontaneous abortion may be related to anatomic, genetic, infectious, immunologic, endocrine, environmental or psychologic problems. A thorough evaluation of the possible etiologies may lead to an explanation, a therapy plan and a successful pregnancy. Psychosocial support is an integral part of both evaluation and treatment. PMID- 8249777 TI - Chronic diarrhea: evaluation and treatment. AB - Chronic diarrhea is defined as the passage of more than 200 g of stool per day for more than three weeks. This condition may result from decreased absorption of gastrointestinal contents or increased fluid secretion into the bowel. Although chronic diarrhea can have many etiologies, irritable bowel syndrome, lactose intolerance, dietary factors, inflammatory bowel disease and colon cancer are the causes most frequently encountered in primary care practice. An orderly work-up, beginning with a complete history an a thorough physical examination, is essential. Whenever possible, treatment should be directed at the underlying cause of the diarrheal condition. If the diarrhea persists and the etiology remains obscure, administration of opiates or bile-sequestering agents often is helpful in alleviating symptoms. New approaches to decreasing secretions, such as the use of clonidine therapy, are being studied. PMID- 8249778 TI - Pulmonary complications of asbestos exposure. AB - Except for benign pleural effusion, asbestos-related pulmonary complications, including asbestosis, malignant mesothelioma and bronchogenic carcinoma, usually occur more than 20 years after exposure. Pleural plaques and pleural thickening serve as markers for asbestos exposure, but they are not associated with an increased risk of malignancy. Clinical criteria for the diagnosis of asbestosis include a reliable history of asbestos exposure, an appropriate interval between exposure and disease detection, radiographic evidence of pulmonary fibrosis, decreased vital capacity and diffusing capacity, and bilateral posterior inspiratory crackles. A lung biopsy is indicated only to rule out other causes of interstitial lung disease. A history of dyspnea, pleuritic chest pain, fatigue, weight loss and pleural effusion in a former asbestos worker is suggestive of mesothelioma. Cigarette smoking greatly increases the risk of lung cancer in asbestos workers. PMID- 8249779 TI - Preventing the spread of viral hepatitis. AB - The most common causes of viral hepatitis, both in the United States and worldwide, are the hepatitis A, hepatitis B and hepatitis C viruses. The similarities and differences of these viral forms provide a basis for developing strategies to prevent the spread of viral hepatitis. Preventive strategies include preexposure and postexposure prophylaxis and universal immunization of all infants against hepatitis B. PMID- 8249780 TI - Succimer: the first approved oral lead chelator. AB - The Centers for Disease Control and Prevention has redefined the threshold of concern for low-level lead toxicity, reducing it from a blood lead level of 25 micrograms per dL (1.21 mumol per L) to a blood lead level of 10 micrograms per dL (0.48 mumol per L), and has recommended universal screening of young children. Succimer (2,3-dimercaptosuccinic acid) is an effective oral lead chelating agent that has been approved for outpatient treatment of children with blood lead levels higher than 45 micrograms per dL (2.17 mumol per L). In the United States, clinical experience with succimer is limited; however, observed side effects, including gastrointestinal symptoms, rash and transient elevations of serum aminotransferase levels, are uncommon and mild. Isolated cases of neutropenia have been reported. Weekly monitoring of complete blood counts and serum aminotransferase levels is recommended during the 19-day treatment. Blood lead levels should be checked weekly to identify rebound from bone and soft tissue mobilization. PMID- 8249781 TI - AHA updates guidelines for pediatric basic and advanced life support. PMID- 8249782 TI - Assessment of perfused left ventricular mass in normal, ischemic, and reperfused myocardium by means of single-photon emission computed tomography of technetium 99m isonitrile. AB - To test the hypothesis that single-photon emission tomography of technetium (Tc) 99m hexakis 2-methoxyisobutyl isonitrile (Tc-MIBI) can accurately measure perfused left ventricular (LV) mass in nonischemic, ischemic, and reperfused myocardium, we acquired Tc-MIBI tomographic images in canines with normally perfused hearts (n = 33) after occlusion of the left anterior descending coronary artery (n = 15), after reperfusion (n = 13), and with subsequent second injection of Tc-MIBI (15 to 18 mCi; n = 12). In all ischemic studies the initial dose of Tc MIBI (5 to 6 mCi) was injected after coronary artery occlusion but before reflow. Scintigraphic perfused LV mass was calculated from the total voxels demonstrating Tc-MIBI uptake x voxel volume (cm3) x specific gravity of myocardium (1.05 gm/cm3). After being imaged the animals were killed, the left ventricle was weighed, and the risk area was determined by dual perfusion with phthalocyanine blue dye and triphenyltetrazolium chloride (TTC). Perfused LV mass was defined as total LV mass minus the risk area mass. There was good correlation between scintigraphic and morphologic determinations of perfused left ventricular mass in nonischemic hearts (Tc-MIBI left ventricular distribution = 0.84 x left ventricular weight + .20.4, n = 33, r = 0.93, p = 0.0001) and ischemic hearts (Tc MIBI left ventricular distribution = 0.51 x left ventricular weight + 37.9, n = 15, r = 0.83, p = 0.0001). In animals imaged both before and after reperfusion, scintigraphic determinations of the nonischemic region correlated closely (after reflow Tc-MIBI distribution = 1.07 x before-reflow Tc-MIBI distribution--8.0, n = 13, r = 0.88, p = 0.0001), indicating that Tc-MIBI does not significantly redistribute into the ischemic zone after reperfusion. After injection of the second dose of Tc-MIBI in acutely reperfused canines, there was good correlation between the distribution mass of Tc-MIBI and the mass of viable myocardium by TTC staining (Tc-MIBI distribution = 0.61 x viable LV mass + 34.2, n = 12, r = 0.77, p = 0.0001). Furthermore, the apparent redistribution of myocardial Tc-MIBI from before and after second injection images correlated with the degree of myocardial salvage by histochemical staining (r = 0.72, p = 0.0082). In conclusion, single photon emission computed tomography of Tc-MIBI can measure perfused LV mass accurately in both ischemic and nonischemic canine preparations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8249783 TI - Long-term dietary fish oil supplementation protects against ischemia-reperfusion induced myocardial dysfunction in isolated rat hearts. AB - Dietary fish oil has been shown to exert protective effects against arrhythmias and myocardial infarction after coronary artery occlusion. However, the effects of fish oil on ischemia-reperfusion-induced cardiac dysfunction are not known. This study was designed to examine if long-term dietary fish oil protects against a rise in coronary perfusion pressure and myocardial contractile dysfunction following ischemia and reperfusion. Fifteen Sprague-Dawley rats (age 7 to 9 weeks) were fed fish oil-rich chow for 4 to 5 weeks and 11 rats from the same batch were fed ordinary chow. Three fish oil-fed rats were also fed ad libitum indomethacin for the last 2 days. Isolated hearts from both groups were perfused on a Langendorff apparatus and were subjected to 25 minutes of global ischemia and 20 minutes of reperfusion. Myocardial phospholipid acid content was also measured. After 4 to 5 weeks of dietary fish oil supplementation, myocardial content of long-chain polyunsaturated fatty acid (PUFA) (C20-C22) and omega-3 PUFA was increased and that of omega-6 PUFA was decreased in the fish oil-fed group (all p < 0.01). Following global ischemia and reoxygenation, there was a reduction in the force of cardiac contraction and an increase in coronary perfusion pressure. However, reduction in the force of cardiac contraction was less in the hearts of fish oil-fed rats than in the control hearts (49 +/- 9% vs 63 +/- 5%, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249784 TI - Adenosine deaminase inhibition is not cardioprotective in the rat. AB - Adenosine receptor activation has been assumed to play a role in the cardioprotective effect of ischemic preconditioning. The actions of adenosine are terminated by the naturally occurring substance adenosine deaminase. We determined whether 2'-deoxycoformycin (DCF), a potent inhibitor of adenosine deaminase, could mimic the effect of preconditioning in nonpreconditioned rats and potentiate the salutary effect of preconditioning in preconditioned rats. We assessed the effect of DCF on myocardial infarct size and the incidence of ventricular arrhythmias in four groups of anesthetized rats: control (nonpreconditioned) + vehicle, control + DCF, preconditioned + vehicle, and preconditioned + DCF. All rats underwent 90 minutes of coronary artery occlusion followed by 4 hours of reperfusion, while preconditioned rats received three cycles of 3-minute episodes of ischemia and 5 minutes of reperfusion before the 90-minute occlusion. Following 4 hours of reperfusion, area at risk was determined by intravenous injection of blue dye during a brief coronary occlusion, and area of necrosis was determined by incubation of heart slices in triphenyltetrazolium chloride. In the nonpreconditioned control rats receiving vehicle, myocardial infarct size expressed as a percentage of the area at risk averaged 44.8 +/- 7.6%. Pretreatment with DCF had no effect on infarct size (49.4 +/- 4.9%) in the nonpreconditioned control rats. Both the preconditioned+vehicle (19.2 +/- 7.8%) and the preconditioned + DCF (17.9 +/- 5.1%) groups had a significant reduction in infarct size (p < 0.05 versus control + vehicle and control + DCF), with no significant difference in infarct size between the two preconditioned groups. The incidence of ventricular tachycardia (VT) during the 90 minutes of ischemia was significantly attenuated in both preconditioned groups (p < 0.05 versus controls).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249785 TI - Elevated plasma interleukin-6 levels in patients with acute myocardial infarction. AB - Interleukin-6 (IL-6) plays a key role in the synthesis of human acute-phase protein and several acute-phase responses occur in patients with acute myocardial infarction (AMI). We examined the plasma levels of IL-6 in 23 consecutive patients with AMI over the course of 4 weeks and in 30 control subjects. In patients with AMI, the plasma IL-6 levels (in picograms per milliliter) were increased at all sampling points from admission to discharge (ranging from 28.5 +/- 6.6 to 46.5 +/- 7.8) compared with levels in control subjects (11.4 +/- 2.9; p < 0.01). Cardiac catheterization did not influence plasma IL-6 levels. The plasma IL-6 level reached its peak approximately 3 days (46.5 +/- 7.8) and approximately 1 week after admission in patients with AMI. There was a significant positive linear correlation between the peak level of plasma IL-6 minus the level on admission and the peak level of plasma C-reactive protein in patients with AMI. The peak IL-6 level did not correlate with the peak levels of creatine kinase, pulmonary capillary wedge pressure, or left ventricular ejection fraction at 4 weeks. We conclude that the plasma IL-6 level is increased over a time course of 4 weeks in patients with AMI. PMID- 8249786 TI - Role of coronary vasospasm in the pathogenesis of myocardial infarction: study in patients with no significant coronary stenosis. AB - The role of coronary vasospasm in the pathogenesis of myocardial infarction is unclarified. Among 212 patients with myocardial infarction in whom percutaneous transluminal coronary angioplasty (PTCA) or coronary thrombolysis was not performed at the acute stage, 21 patients (10%) showed no significant coronary stenosis (the degree of stenosis was less than 50% of the luminal diameter) by coronary angiography 4 weeks after myocardial infarction. Among them, 11 (52%) had preinfarction angina at rest, including two with variant angina, and nine (43%) had postinfarction angina at rest. Intracoronary ergonovine maleate induced coronary vasospasm in 12 (75%) of 16 patients examined. Coronary vasospasm occurred in the infarct-related coronary arteries in all patients, and importantly, multivessel coronary vasospasm occurred in 11 patients (69%). The infarct size was relatively small in these patients: (1) seven patients (33%) had Q wave myocardial infarction while 14 patients (67%) had non-Q wave myocardial infarction; (2) peak creatine phosphokinase (CPK) was lower than 1000 IU/ml in all patients; and (3) thallium-201 (Tl-201) scintigraphic study showed no perfusion defect in 8 of 18 patients. There was only one patient with congestive heart failure and no patient died. These results suggest that coronary vasospasm may play an important role in the pathogenesis of myocardial infarction in patients without significant coronary stenosis. The relatively small infarct size suggests that coronary reperfusion occurred in the early stages of myocardial infarction. PMID- 8249787 TI - Risk factors and mortality among black, Caucasian, and Latina women with acute myocardial infarction. AB - The purpose of this study was to examine the clinical outcome of acute myocardial infarction among women of three ethnic/racial groups and to determine whether observed differences in outcome are explained by differences in associated risk factors. Nine hundred seventeen (917) consecutive admissions occurred among 810 women, of whom 347 (42.8%) were Caucasian, 258 (31.9%) were black, and 205 (25.3%) were Latina. The overall mortality rate was 13%; the Caucasian rate was significantly higher than the Latina rate (16.9% vs 7%, p < 0.01), as was the black rate (13.8% vs 7%, p < 0.05). Comparing survivors with nonsurvivors among the three groups, hypertension was more common for survivors among blacks, as was a history of angina; in-hospital congestive failure was associated with a higher mortality for blacks and Caucasians, as was in-hospital angina for Caucasians. Univariate and multivariate logistic regression for the outcome variables of congestive failure, in-hospital angina, and mortality, showed race to be a significant factor in the lower mortality rate for Latinas in the univariate analysis. In the multivariate analysis, only in-hospital angina and congestive failure significantly influenced mortality. In this study of myocardial infarction among women of lower socioeconomic status, factors other than race were the primary influences on outcome. PMID- 8249788 TI - Exercise-induced QRS prolongation in patients with coronary artery disease: a marker of myocardial ischemia. AB - The objective of this study was to investigate the effect of myocardial ischemia on the QRS duration in patients with coronary artery disease since acute myocardial ischemia decreases conduction velocity through the ischemic myocardium and may produce QRS prolongation on the surface electrocardiogram. One hundred fifty patients who underwent cardiac catheterization and exercise radionuclide ventriculography within 1 month of each other were studied. Forty patients had normal coronary arteries and 110 had coronary artery disease. QRS duration decreased with exercise in patients with normal coronary arteries (-3.0 msec, confidence limits -6.2 to 0.2), but increased in patients with coronary artery disease; exercise-induced QRS prolongation was directly related to the number of diseased vessels (4.8 msec in patients with one, 7.8 msec in patients with two, and 13.3 msec in patients with three-vessel disease, p < 0.001). Likewise, QRS duration decreased with exercise in patients without exercise-induced segmental contraction abnormalities (-1.8 msec, confidence limits -3.7 to 0.1), but increased in patients with segmental contraction abnormalities (6.7 msec in patients with one, 13.5 msec in patients with two, and 21 msec in patients with three segmental contraction abnormalities, p < 0.0001). Exercise-induced QRS prolongation was better related to the number of segmental contraction abnormalities than to the number of diseased vessels (p < 0.01). It was concluded that exercise produces QRS prolongation in patients with coronary artery disease in direct relation to the number of diseased vessels and to exercise-induced segmental contraction abnormalities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249789 TI - Quantitative angiography during coronary angioplasty with a single angiographic view: a comparison of automated edge detection and videodensitometric techniques. AB - Little information is available on the reliability of coronary luminal measurements obtained from quantitative analysis of a single angiographic view, an approach that is central to the practical use of on-line quantitative angiography. In the present study we investigated the contribution of two different techniques of quantitative angiography, edge detection (ED) and videodensitometry (VD), to the application of this concept during coronary angioplasty. Forty-six balloon angioplasty procedures were included in this study, all of them performed in a stenosis located in the mid right coronary segment. This coronary location was chosen to optimize data collection on luminal morphology and to minimize the number of factors that may adversely affect quantitative analysis with both techniques. In all cases two orthogonal angiographic projections were obtained before, after balloon dilatation, and at follow-up. Correlation coefficients and differences between orthogonal measurements obtained with each technique were used to evaluate the agreement between orthogonal readings at every stage of the procedure. The obtained correlation coefficients and mean differences (MD) between orthogonal measurements were as follows: before percutaneous transluminal coronary angiography (PTCA), 0.67 (MD 0.01 +/- 0.47 mm2) and 0.57 (MD 0.05 +/- 0.64 mm2) for ED and VD, respectively (Pitman's test for SD, p < 0.05); after balloon dilatation, 0.32 (MD -0.56 +/- 1.53 mm2) and 0.53 (MD -0.15 +/- 1.43 mm2) for ED and VD, respectively (paired t test for MD, p < 0.05); and at follow-up 0.79 (MD 0.15 +/- 0.97 mm2) and 0.73 (MD 0.17 +/- 1.16 mm2) for ED and VD, respectively (p = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249790 TI - Coronary restenosis: evaluation of a restenosis injury index in a swine model. AB - To investigate the mechanisms of restenosis and detect useful interventions to prevent it, reliable quantitative measurements must be evaluated. Coronary arteries of domestic and minipigs (n = 18) were mechanically injured by balloon overstretching and killed at different intervals (2 to 25 weeks) after quantitative angiographic analysis. Morphometric measurements evaluated intimal hyperplasia at 0.59 +/- 0.42 mm without relation to artery size or balloon/artery ratio. Intimal hyperplasia, expressed as the ratio of neointimal area to total wall area (A), is directly related to the injury, assessed by the ratio of internal elastic lamina (IEL) fracture length to IEL circumference (B), r = 0.84, p = 0.002. Restenosis injury index, defined as A/B, provides a useful tool for the quantitative assessment of future angioplasty-related restenosis interventions. PMID- 8249791 TI - Clinical features and electrophysiologic characteristics of accessory atrioventricular pathways and atrioventricular nodal reentrant tachycardia: comparative study between young and elderly patients. AB - Information focused on elderly patients with paroxysmal supraventricular tachycardia, and a comparative study between the elderly and younger patients was limited. The aim of this study was to investigate the clinical features and electrophysiologic characteristics in elderly patients (> or = 65 years of age) with accessory atrioventricular pathway-mediated tachyarrhythmia or atrioventricular nodal reentrant tachycardia. Electrophysiologic studies of 45 elderly patients with accessory pathway-mediated tachyarrhythmia and 47 elderly patients with atrioventricular nodal reentrant tachycardia were performed. When compared with younger patients, the elderly patients had a similar incidence of critical clinical manifestation, higher incidence of other cardiac disease or systemic disease, similar inducibility of tachycardia, greater baseline electrophysiologic parameters (sinus node, atrioventricular node, atrium and ventricle), and poorer properties of accessory pathways and dual atrioventricular nodal pathways. These results suggest that conduction properties of accessory pathways and dual atrioventricular nodal pathways were poorer in elderly patients, but similar critical manifestations and tachycardia events were found in elderly and younger patients. PMID- 8249792 TI - Spectrum of right heart involvement in patients with ventricular tachycardia unrelated to coronary artery disease or left ventricular dysfunction. AB - We studied 41 patients with clinically symptomatic ventricular tachycardia in the absence of coronary artery disease or left ventricular dysfunction to define the extent of right and left heart derangement and their relation to electrophysiologic and clinical data. Individual echocardiographic measurements as well as global assessment scores of the right and left heart demonstrated a wide spectrum of right heart echocardiographic abnormalities. There was much less variation in the left heart, with the majority of patients being close to normal. There was an association between the right heart score, the clinical presentation of arrhythmia, the response to programmed electrical stimulation, and the recurrence of arrhythmia (p < 0.05). Thus echocardiographic findings demonstrate the whole spectrum of right heart involvement in patients with apparent idiopathic ventricular tachycardia and can give insight into clinical history, arrhythmia inducibility, and prognosis. PMID- 8249793 TI - Dynamics of the QT interval in patients with exercise-induced ventricular tachycardia in normal and abnormal hearts. AB - Inhomogeneity of ventricular repolarization reflected in prolongation of the QT interval of the surface electrocardiogram can predispose patients to ventricular arrhythmia. This study examines whether an abnormality of QT adaptation to changes in heart rate is likely to be of importance in the pathogenesis of ventricular tachycardia (VT) in patients with and without underlying structural heart disease. The QT-R-R relationship during exercise was studied in 52 patients. Forty-two patients had VT associated with a "clinically normal" heart (idiopathic VT), of which 23 had no VT on exercise and 19 had exercise-induced VT. These patients were compared to 10 subjects with exercise-induced VT related to ischemic heart disease. The QT interval was measured manually from computer averaged QRS complexes recorded at 1- to 3-minute intervals during treadmill exercise tests. An approximately linear association existed between the QT and R R intervals within the range of heart rates observed. The slope of the QT-R-R relation was lower in patients with structural heart disease (0.23 +/- 0.06) than in patients with normal hearts with (0.29 +/- 0.12) and without (0.29 +/- 0.12) exercise-induced VT (p < 0.05). The intercept of the regression line was higher in patients with structurally abnormal hearts (209.2 +/- 55.3 msec) than in patients with idiopathic VT with (155.6 +/- 49.7 msec) and without (157.7 +/- 69.0 msec) exercise-induced VT (p < 0.02). The corrected QT (Bazett's formula) was similar all three groups at rest, but was higher in patients with structurally abnormal hearts at peak exercise, 449.6 +/- 28.0 versus 425.8 +/- 27.4 msec (idiopathic VT, exercise induced) versus 427.3 +/- 26.6 msec (idiopathic VT, not exercise induced) (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249794 TI - Alterations in heart rate variability and its circadian rhythm in hypertensive patients with left ventricular hypertrophy free of coronary artery disease. AB - Heart rate variability (HRV) and its circadian rhythm were evaluated in 22 patients with treated hypertension and left ventricular hypertrophy in whom coronary artery disease was excluded by stress thallium or angiography. By using 24-hour Holter monitoring, HRV and its spectral components were measured. Findings were compared with 11 age-matched normal controls. The difference between mean R-R intervals during sleep (11 PM to 7 AM) and while awake (9 AM to 9 PM) (73 +/- 33 vs 263 +/- 63 msec, p < 0.0001) and the mean 24-hour SD of the R R intervals (55 +/- 6.3 vs 93 +/- 11, p < 0.0001) were lower among the hypertensive patients compared with controls. The percentage of difference between successive R-R intervals that exceeded 50 msec, a measure of parasympathetic tone, was also lower among the hypertensive patients (6.8 +/- 7.1 vs 13.6 +/- 8.9, p < 0.002); it increased at night and decreased during the day among the controls, and this circadian rhythm was blunted among the patients. Spectral analysis showed that power in the high-frequency range (0.15 to 0.40 Hz) was lower among the hypertensive patients during 21 of 24 hours but that the difference was statistically significant only during 9 hours (p ranging from < 0.05 to 0.009). Power in the low-frequency range (0.04 to 015 Hz) was lower at night, increased in the morning, and higher during the day among controls; this circadian rhythm was absent among hypertensive patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249795 TI - Evaluation of left ventricular mass: comparison of ultrafast computed tomography, magnetic resonance imaging, and contrast left ventriculography. AB - We measured and compared left ventricular mass in 20 patients by ultrafast computed tomography (UFCT), magnetic resonance imaging (MRI), and contrast left ventriculography (LVG). Left ventricular mass was calculated by UFCT and MRI in two ways: (1) excluding papillary muscles and trabeculae (LV mass), and (2) including papillary muscles and trabeculae (LV mass + PM&T) by Simpson's method. Left ventricular mass excluding papillary muscles and trabeculae (LV mass) in LVG was calculated by Rackley's method by biplane angiocardiography. LV mass was significantly larger in LVG than in MRI and UFCT (p < 0.01). Although LV mass was significantly larger in MRI than in UFCT (p < 0.01), there was no significant difference in LV mass + PM&T between UFCT and MRI. Interobserver and intraobserver variability showed good correlation of coefficient in both UFCT and MRI. We therefore conclude that left ventricular mass is best measured by including papillary muscles and trabeculae by Simpson's method in UFCT or MRI. PMID- 8249796 TI - Magnetic resonance imaging compared with angiography in the evaluation of intermediate-term result of coarctation balloon angioplasty. AB - Between July 1986 and December 1990, 24 consecutive adult patients with native coarctation of the aorta underwent balloon dilatation. Their ages ranged from 15 to 55 (mean 25) years. Dissection of the aorta developed in one patient. The remaining 23 patients were restudied by catheterization and magnetic resonance imaging (MRI) 8 to 60 (mean 21) months after dilatation. Both studies were performed between 1 and 180 (mean 40) days of each other. The diameter of the aorta at the site of previous coarctation was measured on angiogram and MRI by two independent observers. The data were compared by means of linear regression analysis. The gradient across the previous coarctation site ranged from 0 to 20 (mean 7 +/- 7.3) mm Hg. The diameter of the aorta at the site of previous coarctation measured on angiogram was 13.7 +/- 3.7 mm and on MRI it measured 13.5 +/- 3.7 mm, with excellent correlation (r = 0.96, SEE = 0.92, p < 0.001). Two patients had small aneurysms 2 cm in diameter demonstrated by angiography and MRI, and two patients developed restenosis, diagnosed correctly by both cardiac catheterization and MRI. This study demonstrates that MRI provides excellent visualization of the anatomy of the aorta and is a good noninvasive method for follow-up of patients undergoing balloon coarctation angioplasty. PMID- 8249797 TI - Immunohistochemical localization of laminin in the hearts of patients with chronic chagasic cardiomyopathy: relationship to thickening of basement membranes. AB - Histologic, ultrastructural, and immunohistochemical studies were made of endomyocardial biopsy specimens from 10 patients with chronic chagasic cardiomyopathy. Histologic and electron microscopic observation disclosed marked thickening of the basement membranes of the myocytes, endothelial cells, and vascular smooth muscle cells in all patients. Light (peroxidase-labeled antibodies) and electron (gold-conjugated antibody) microscopic immunohistochemical methods revealed a positive reaction for laminin in these thickened basement membranes. This thickening of basement membranes may develop as a consequence of: (1) an immunologic reaction that is triggered by the presence of a laminin-like molecule on the surfaces of Trypanosoma cruzi amastigotes and trypomastigotes; (2) an immunologic response to direct injury of basement membranes causing some of their components to become antigenic; (3) myocardial fibrosis, with synthesis of new connective tissue components; and (4) a combination of the preceding factors. The relationship of these changes to antilaminin antibodies requires clarification. PMID- 8249798 TI - Relation of pulmonary venous flow to mean left atrial pressure in mitral stenosis with sinus rhythm. AB - To determine whether pulmonary venous flow measured by transesophageal Doppler echocardiography can be used to estimate mean left atrial pressure (LAP), we prospectively studied 12 consecutive patients with sinus rhythm undergoing percutaneous mitral balloon commissurotomy for their severe mitral stenosis (mitral valve area < 1.5 cm2). We correlated Doppler variables of pulmonary venous flow and the mean LAP measured by left atrial catheterization. Among the variables of the pulmonary venous flow, the systolic fraction (i.e., the systolic velocity-time integral expressed as a fraction of the sum of systolic and early diastolic velocity-time integral) correlated significantly with mean LAP (r = 0.71, p < 0.05) and mitral valve area (r = 0.64, p < 0.05). Peak velocity and velocity-time integral in systole also significantly correlated with mean LAP (r = -0.66, r = -0.67 respectively, p < 0.05). We conclude that the more severe the degree of mitral stenosis in patients with sinus rhythm, the less systolic pulmonary venous flow in severe mitral stenosis. PMID- 8249799 TI - Doppler echocardiographic assessment of left ventricular filling in idiopathic dilated cardiomyopathy during a one-year follow-up: relation to the clinical course of disease. AB - In idiopathic dilated cardiomyopathy (IDC), an impaired left ventricular filling as assessed by the Doppler echocardiographic mitral flow pattern is closely related to the severity of congestive heart failure. This study examined the relation of left ventricular filling and the clinical course of the disease in patients with a recent diagnostic procedure and initiation of medical therapy (group 1, n = 15) as compared with patients in a chronic stage of the disease (group 2, n = 24) with the diagnosis established > 1 year before. All patients had to be in sinus rhythm to facilitate the Doppler echocardiographic evaluation of left ventricular filling. The clinical status was assessed by the New York Heart Association classification and a heart failure score at baseline and after a period of 12 +/- 7 months. At baseline the ratio of the peak early/atrial Doppler velocities (VE/VA) was shifted toward the early diastole in group 1 as compared to group 2 (1.84 +/- 1.02 vs 1.12 +/- 0.55; p < 0.05). Symptoms of heart failure were more severe in group 1. During follow-up, VE/VA tended to decrease in group 1 from 1.84 +/- 1.02 to 1.35 +/- 1.03 (p = 0.07) and remained unchanged in group 2 (1.12 +/- 0.55 and 1.34 +/- 1.23; not significant). In a subgroup of 10 patients who underwent repeat right heart catheterization, the decrease of VE/VA coincided with a decrease of the pulmonary capillary wedge pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249800 TI - Abnormal changes in transmitral flow after acute exposure to nitroglycerin and nifedipine in uncomplicated insulin-dependent diabetes mellitus: a Doppler echocardiographic study. AB - To study the left ventricular (LV) diastolic function in uncomplicated insulin dependent diabetic patients without hypertension (n = 25) and comparable controls (n = 15), the effect of acute administration of nitroglycerin and nifedipine on cavity dimensions and transmitral flow pattern was investigated by Doppler echocardiography. At baseline no significant differences in any of the LV Doppler echocardiographic parameters were seen in the two groups. Only in diabetics did nitroglycerin diminish LV diastolic and systolic diameter significantly (p < 0.0002 and 0.004, respectively), reducing the stroke volume from 70 +/- 16 to 64 +/- 20 ml, p < 0.01. The decrease in stroke volume correlated significantly to hemoglobin (Hb) A1c level (r = 0.42, p = 0.036). An excessive preload reducing effect on venous capacitance vessels is assumed in diabetes, and this preload reducing effect was also reflected in transmitral flow pattern where E-wave/A wave ratio decreased significantly only in the diabetic subjects (p < 0.0005). Nifedipine induced the same degree of sympathetic activation in the two groups, but an increase in LV early peak filling rate (E wave) was seen in diabetes only (63.3 +/- 13.5 to 66.8 +/- 13.5 mm, p < 0.01). The atrial filling of LV (A wave) was increased significantly in both groups. Thus an abnormal LV diastolic function can be disclosed by these pharmacologic challenges. Regulation of preload and its influence on transmitral flow pattern in diabetes deserves further investigation, as does the influence of calcium antagonists on early relaxation in the diabetic myocardium. PMID- 8249801 TI - Diagnostic considerations in patients with suspected coronary artery anomalies. AB - The diagnosis of coronary artery anomalies requires a high index of suspicion during the history and physical examination. Nonspecific presentations are common, although dramatic presentations such as myocardial infarction or aborted sudden death will occur. Typically the results of the physical examination and ECG and the chest x-ray film are normal except with ALCAPA or coronary-artery fistulas. Screening studies such as exercise stress testing have low sensitivity, and a negative evaluation does not rule out the presence of a significant anomaly. These important limitations must be considered. Echocardiography is a practical and frequently diagnostic test if specific attention is paid to the coronary arteries. This examination should be performed in most patients with suspect symptoms. Other noninvasive techniques such as nuclear and cine MRI cardiac imaging may become increasingly important, but their current use is anecdotal. Cardiac catheterization remains the gold standard; however, recognition of important clues and specific angiographic views are required to fully delineate many anomalies. PMID- 8249802 TI - Magnetic resonance measurement of velocity and flow: technique, validation, and cardiovascular applications. AB - With a newly developed magnetic resonance (MR) technique for blood flow measurements, qualitative and quantitative information on both flow volume and flow velocity in the great vessels can be obtained. MR flow quantitation is performed with a gradient-echo MR sequence with high temporal resolution enabling measurements at frequent intervals throughout the cardiac cycle. MR flow quantitation uses the phase rather than the amplitude of the MR signal to reconstruct the images. These images, often referred to as MR velocity maps or velocity-encoded cine MR images, are two-dimensional displays of flow velocity. From these velocity maps, velocity and volume flow data can be obtained. Previous validation experiments have demonstrated the accuracy of MR velocity mapping, and this technique is now being applied successfully in several clinical fields. MR velocity mapping may be of considerable value when Doppler echocardiography results are unsatisfactory or equivocal, particularly because MR is suited for the analysis of volumetric flow and complex flow patterns. Among the vastly growing number of clinical cardiovascular applications that have been reported are the great arteries and veins, coronary vessels, valvular disease, and the abdominal and peripheral vessels. These items are reviewed, and some aspects of the technique that need improvement are discussed. PMID- 8249803 TI - Validation of volumetric flow measurements by means of a Doppler-tipped coronary angioplasty guide wire. AB - We used an in vitro model to validate volumetric flow measurements obtained with an 0.018-inch angioplasty guidewire with a 12 MHz transducer mounted on its tip. By using a modified two-head roller pump device, flow was adjusted incrementally from a minimum of 90 ml/min to a maximum of 550 ml/min. Flow was measured with the Doppler guide wire in tubing ranging from 1.9 mm to 6.0 mm internal diameter, as the product of the spectral Doppler velocity integral and the cross-sectional area of the tubing, over a 1-minute period. It was an excellent correlation between the Doppler calculated flow rates and actual flow, regardless of tubing diameter (r = 0.99). These results suggest that the Doppler spectral output of this device might be accurately applied to estimates of volumetric flow in human coronary arteries. PMID- 8249804 TI - Peripartum myocardial infarction. PMID- 8249805 TI - Coronary subclavian steal syndrome. PMID- 8249806 TI - Percutaneous transluminal angioplasty of the innominate artery combined with carotid endarterectomy. PMID- 8249807 TI - Regression of a dilated cardiomyopathy after radiofrequency ablation of incessant supraventricular tachycardia. PMID- 8249808 TI - Idiopathic ventricular fibrillation and bradycardia-dependent intraventricular block. PMID- 8249809 TI - Ablation and coagulation of myocardial tissue by means of a pulsed holmium:YAG laser. PMID- 8249810 TI - Feasibility and safety of pulsed dye laser irradiation in the pulmonary arteries. PMID- 8249811 TI - Aortico-left ventricular tunnel with aortic atresia in the newborn. PMID- 8249812 TI - Transesophageal echocardiographic diagnosis of right atrioventricular valve patch dehiscence causing intracardiac right-to-left shunting after Fontan operation. PMID- 8249813 TI - Plasma endothelin and pulmonary pressures in patients with congestive heart failure. PMID- 8249814 TI - Endothelial dysfunction after reperfusion. PMID- 8249815 TI - CNS-mediated coronary spasm. PMID- 8249816 TI - Low-nicotine cigarettes on CAD. PMID- 8249817 TI - Systemic embolization during TTE. PMID- 8249818 TI - Balloon dilation of aortic coarctation. PMID- 8249819 TI - Doppler-detected aortic coarctation with PDA. PMID- 8249820 TI - Umbilical vein cardiac catheterization. PMID- 8249821 TI - Diltiazem myopathy. PMID- 8249822 TI - Arterial thrombosis in thrombocythemia. PMID- 8249823 TI - Chagas' disease in north America. PMID- 8249824 TI - U.S. trends in rheumatic fever and heart disease. PMID- 8249825 TI - Cardiac allograft vasculopathy. PMID- 8249826 TI - A sample of dispensing errors reported. PMID- 8249827 TI - Flu vaccinations. PMID- 8249828 TI - Assessing and counseling contact lens wearers. PMID- 8249829 TI - Estrogens for treating urinary incontinence in women. PMID- 8249830 TI - Comparing professional and consumer drug information centers. AB - Providing drug information to consumers is an essential component of medical and pharmaceutical care. In recent years, profit-based consumer-oriented drug information centers (DICs) or drug hot lines have developed to fill this need. This study compared the services and information provided by DICs for consumers with those provided by DICs for health care professionals. The two consumer oriented DICs provided-accurate and professional information, similar in content to that provided by the health care professional DICs. Although the data were too small for statistical analysis, it appears that the consumer-oriented DICs offer a valuable service. PMID- 8249831 TI - Drug information center for community pharmacists. PMID- 8249832 TI - HIV infection: a pharmacist's resource. APhA Editorial Board on Pharmaceutical Care for Patients with HIV Infection. PMID- 8249833 TI - Lead poisoning. PMID- 8249834 TI - Health care reform and PAID's network. PMID- 8249835 TI - Initial clinical presentation of cardiac disease in asymptomatic men with silent myocardial ischemia and angiographically documented coronary artery disease (the Oslo Ischemia Study). AB - Data concerning the natural history of asymptomatic coronary artery disease (CAD) has been limited to epidemiologic rather than angiographic studies, thus leading to uncertainty as to whether warning symptoms and signs will identify subjects with silent myocardial ischemia before morbid events. To address this issue, 50 apparently healthy men with angiographically proven CAD and asymptomatic exercise induced ST depression have been followed prospectively for 15 years in the Oslo Ischemia Study. Fourteen men died. The initial presenting clinical event in these 14 men was chest pain in 4 (30%)--but in only 1 case was it recognized as typical angina--silent myocardial infarction in 5 (35%) and sudden death in 5 (35%). Thirty-six men survived, with 19 developing symptoms. Overall, chest pain was the first clinical event in 22 of the total of 33 men with symptoms (66%), whereas myocardial infarction occurred in 6 (18%) and sudden death in 5 (16%). Although chest pain occurred in 22 men, it was clinically diagnosed as typical angina pectoris in only 6. These observations suggest that there is an absence of clear cut ischemic symptoms in many asymptomatic patients before morbid events. PMID- 8249836 TI - Late outcome after percutaneous transluminal coronary angioplasty during acute myocardial infarction. AB - Early reperfusion for acute myocardial infarction (AMI) results in improved ventricular function and survival. There is a dearth of data on long-term survival (> 5 years) after percutaneous transluminal coronary angioplasty (PTCA) performed either as a primary procedure or in conjunction with thrombolytic therapy. We studied 160 patients who underwent PTCA during AMI between 1981 and 1987 either with (n = 101) or without (n = 59) streptokinase therapy. Mean time to reperfusion was 4.6 hours, and patency was achieved in 134 patients (84%). Mean discharge ejection fraction was 46 +/- 14%. Coronary artery bypass grafting was performed before dismissal in 34 patients (21%), including 21 of 130 patients (16%) with 1- or 2-vessel disease and 13 of 30 patients (43%) with 3-vessel disease (p < 0.05). Eleven patients (7%) died in the hospital. The 149 hospital survivors were followed for a mean of 69 +/- 21 months (median 72). During follow up, 22 patients (15%) died, 21 (14%) had reinfarction, 23 (15%) underwent coronary artery bypass grafting, and 21 (14%) underwent repeat PTCA of the infarct-related artery. On univariate analysis, age > or = 62 years, multivessel disease, ejection fraction < or = 40%, previous AMI, and being a nonsmoker at the time of AMI were predictive of late mortality (p < 0.05 each variable). On multivariate analysis, only ejection fraction < or = 40% and prior AMI were predictive of late death. In patients treated with PTCA for AMI, late survival is excellent. Early surgical revascularization of high-risk patients may contribute to these encouraging results. PMID- 8249837 TI - Digital dipyridamole stress echocardiography in silent ischemia after coronary artery bypass grafting and/or after healing of acute myocardial infarction. AB - This study evaluates dipyridamole stress echocardiography in silent ischemia. Fourteen patients with previous coronary artery bypass grafting (group A) and 16 patients with healed myocardial infarction (group B) were studied. All had > or = 1 mm ST depression without chest pain during bicycle exercise testing. Left ventricular wall motion was analyzed using a computerized display of digital systolic cineloops with a high frame rate. Test results were compared with coronary angiography. Dipyridamole echocardiography accurately identified patients with significant coronary artery stenosis in both groups (3 of 4 in group A, 11 of 14 in group B). Retrograde flow to the occluded native artery was associated with positive results on dipyridamole testing in 6 of 7 patients in group A and all 3 in group B. Sensitivity, specificity and diagnostic accuracy for detecting significant coronary stenosis or occlusions with retrograde flow was 78, 100 and 83%, respectively. Patients with angiographic multivessel disease had a significantly larger increase in wall motion score index during dipyridamole stress than patients with 0- or 1-vessel disease, 0.18 +/- 0.11 versus 0.05 +/- 0.18 (p < 0.05). Two patients developed symptomatic bradycardia and hypotension during dipyridamole infusion. It is concluded that dipyridamole echocardiography accurately identifies myocardial regions with restricted coronary flow. Stress echocardiography is a valuable tool for assessing coronary flow in silent ischemia. PMID- 8249838 TI - Effects of streptokinase during acute myocardial infarction on the signal averaged electrocardiogram and on the frequency of late arrhythmias. AB - Although a number of studies have shown that the incidence of late potentials is lower after thrombolytic therapy, it is not known whether this is paralleled by fewer arrhythmic events during long-term follow-up. In patients with first acute myocardial infarction, filtered QRS duration was significantly shorter when treated with streptokinase (95 +/- 11 ms, n = 53) than when treated with conventional therapy (99 +/- 12 ms, n = 77, p < 0.05). The low-amplitude signal (D40) was shorter after thrombolysis (28 +/- 11 vs 33 +/- 12 ms, p < 0.02). Terminal root-mean-square voltage did not differ significantly (41 +/- 24 vs 35 +/- 23 microV). Irrespective of treatment, late potentials were predictive in the complete group (n = 171) for arrhythmic events during follow-up (13 +/- 6 months, range 6 to 24) (hazard ratio 7.7, p < 0.02, Cox proportional-hazards survival analysis), but treatment (streptokinase vs conventional) did not significantly affect outcome when added to the model. It is concluded that thrombolysis prevents the development of late potentials. However, this study does not confirm the hypothesis that prevention of late potentials leads to a decrease in arrhythmic events. PMID- 8249839 TI - Comparison of the usefulness of the implantable cardioverter-defibrillator and subendocardial resection in patients with sustained ventricular arrhythmias and poor regional wall motion associated with coronary artery disease. AB - The implantable cardioverter-defibrillator (ICD) and subendocardial resection are effective forms of therapy for sustained ventricular arrhythmias associated with coronary artery disease in selected patients. The relative efficacy of these 2 treatments in equivalently matched patients is not known. A regional wall motion score has been shown to be a powerful predictor of long-term outcome after both ICD implantation and subendocardial resection. This study retrospectively analyzed the long-term outcome of patients with coronary artery disease and ventricular arrhythmias treated during the same period with an ICD (n = 53) or by subendocardial resection (n = 65). Treatment outcomes were compared in subgroups determined by preoperative regional wall motion scores of either < or = 16 or > 16%. The 3-year cardiac mortality of the 2 therapies was not significantly different among patients with a wall motion score of > 16% (0% ICD vs 11% endocardial resection) or of < or = 16% (41% ICD vs 35% endocardial resection). Similarly, the 3-year sudden cardiac death mortality was similar among patients with a score of > 16% (0% for both ICD and endocardial resection) or of < or = 16% (9% ICD vs 14% endocardial resection, p = NS). At 24 months after hospital discharge, the percentage of patients who were in New York Heart Association functional class I or II was similar among patients with a wall motion score of > 16% (75% ICD vs 86% endocardial resection, p = NS) or with a wall motion score of < or = 16% (26% ICD vs 45% endocardial resection, p = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249840 TI - Transesophageal echocardiography during radiofrequency catheter ablation of ventricular tachycardia. AB - Radiofrequency lesion formation requires stable catheter tip/endocardial contact. Energy delivery is limited when temperatures are > 100 degrees C due to coagulum formation at the catheter tip. Transesophageal echocardiographic imaging may be useful for monitoring catheter position and detecting boiling. Transesophageal echocardiographic images were recorded during production of 22 radiofrequency lesions in bovine myocardium in a saline bath. Lesion size, tissue temperature and appearance of echo contrast (bubbles) were assessed. In 11 patients, transesophageal echocardiography was used to guide catheter movement and detect boiling during radiofrequency ablation for ventricular tachycardia. In the tissue bath, the appearance of echo bubbles was associated with visual bubbling at the catheter tip, tissue temperatures > 60 degrees C and larger lesions (284 +/- 165 vs 30 +/- 54 mm3; p < 0.001). In humans, transesophageal images easily identified the catheter tip in either ventricle and enabled continuous observation of electrode-tissue contact during radiofrequency application. Transesophageal echocardiographic bubbles appeared in 59 of 217 radiofrequency applications (27%). Continued radiofrequency application after appearance of bubbles was followed by an increase in impedance. Prolonged placement of the probe in heavily sedated patients resulted in a mild sore throat, but no other complications. Transesophageal echocardiographic imaging enables continuous monitoring of catheter position during radiofrequency energy application. The abrupt appearance of echo bubbles indicates boiling and impending coagulum formation at the catheter tip. PMID- 8249841 TI - Echocardiographic assessment of left ventricular structure and diastolic filling in elderly subjects with borderline isolated systolic hypertension (the Framingham Heart Study). AB - Abnormalities in left ventricular (LV) structure and function have been shown in patients with diastolic hypertension and recently in subjects with isolated systolic hypertension. The purpose of this study was to determine whether abnormalities of cardiac structure or function are present in elderly subjects with borderline isolated systolic hypertension (defined as systolic blood pressure [BP] between 140 and 159 mm Hg, and diastolic BP < 90 mm Hg). Ninety-one subjects (mean age 77 years) from the original Framingham Heart Study with untreated borderline isolated systolic hypertension, who were free of cardiovascular disease, were compared with 139 normotensive (BP < 140/90 mm Hg) subjects (mean age 76 years). Measurements included M-mode values for LV structure, and 6 Doppler indexes of LV diastolic filling. Subjects with borderline isolated systolic hypertension and the control group differed in mean systolic (147 vs 125 mm Hg) and diastolic (76 vs 70 mm Hg) BP. Borderline systolic hypertension was the most frequent form of untreated hypertension in this elderly group. The sum of LV wall thicknesses (septum+posterior wall) was significantly higher in borderline hypertensive subjects than in normotensive ones (20.5 vs 19.7 mm; p = 0.002). No difference was detected in LV internal dimension or systolic function. After adjustment for age and other clinical variables, comparisons between the groups revealed significant differences in indexes of Doppler diastolic filling. Peak velocity of early filling, and the ratio of early to late peak velocities were lower in the hypertensive group (40 vs 44 cm/s [p = 0.03] and 0.69 vs 0.76 [p = 0.01], respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249842 TI - Usefulness of piretanide plus ramipril for systemic hypertension: a multicenter trial. AB - To test the dose responses of piretanide, ramipril, and their combination in patients with essential hypertension, a prospective, randomized, double-blind, placebo-controlled trial was conducted in 480 patients. Twelve separate groups were studied: placebo, piretanide 3 mg, piretanide 6 mg, ramipril 2.5 mg, ramipril 5 mg, ramipril 10 mg, and their combinations, as single daily morning doses. Patients were randomized after a 2-week run-in period without drugs; treatment was given for 6 weeks. A dose response compared with placebo was found for both drugs; the combination was more effective than either drug alone. Piretanide 6 mg, combined with ramipril 5 mg, provided optimal blood pressure reduction. Self-reported adverse effects of both drugs and their combinations did not exceed those reported for placebo. A surface analysis suggested that piretanide primarily reduced systolic blood pressure, whereas ramipril was more effective in reducing diastolic blood pressure. The data attest to a combined efficacy of piretanide and ramipril in decreasing arterial blood pressure. PMID- 8249843 TI - Impact of left ventricular cavity size on survival in advanced heart failure. AB - Although left ventricular (LV) dilation has been assumed to be deleterious, the physiologic significance of severe LV dilation in advanced heart failure and its impact on survival have not been defined. LV end-diastolic dimension was measured by M-mode echocardiography in 382 patients with class III or IV heart failure symptoms (mean LV ejection fraction 20 +/- 8%) referred for evaluation for cardiac transplantation. All patients underwent right-sided heart catheterization, and received vasodilator and diuretic therapy adjusted to hemodynamic goals. Although 183 patients with massive LV dilation by LV index > 4 cm/m2 (LV index = LV end-diastolic dimension/estimated body surface area) had a similar severity of hemodynamic impairment to that of 199 patients with only moderate dilation (LV index < or = 4 cm/m2), with baseline mean cardiac index of 2 liters/m/m2 and mean pulmonary arterial wedge pressure of 26 mm Hg in both groups, their actuarial 2-year survival without transplantation was much lower (49 vs 75%; p = 0.004). In the Cox proportional-hazards model, LV index predicted total and sudden death, independent of etiology of heart failure, ejection fraction and other parameters of disease severity. Follow-up echocardiograms (mean 13 +/- 6 months) in 80 heart failure survivors without transplantation showed an increase in mean LV ejection fraction (22 +/- 8% to 26 +/- 13%), but no change in mean LV index in either the massive or moderately dilated groups. Thus, massive LV dilation is an independent contributor to poor outcome in patients with advanced heart failure, and may be stabilized by aggressive vasodilator and diuretic therapy. PMID- 8249844 TI - Diagnostic accuracy of transesophageal echocardiography for detecting left atrial thrombi in patients with rheumatic heart disease having undergone mitral valve operations. AB - Transesophageal echocardiography (TEE) has emerged as an efficient method for detecting left atrial (LA) thrombi in recent years, but its accuracy has not been fully evaluated. A prospective clinicopathologic study in 213 consecutive patients with chronic rheumatic mitral valve disease over a period of 39 months was undertaken. All patients underwent open heart surgery within 3 days after the TEE study. The presence or absence of LA thrombi was confirmed at surgery by direct inspection of the left atrium and proven by histopathologic examination. Of the 213 patients, 147 had predominant mitral stenosis, and the remaining 66 patients had significant mitral regurgitation. Twenty-eight patients had LA thrombi by TEE criteria. These findings were all confirmed at surgicopathologic studies (specificity 100%). However, in 2 patients, LA thrombi were present but could not be detected by TEE (sensitivity 93.3%). Therefore, the positive predictive value was 100%, the negative predictive value was 98.9% and the diagnostic accuracy was 99.1%. No thrombi were found in patients with significant mitral regurgitation. The frequency of LA thrombi in patients with predominant mitral stenosis was 20% (30 of 147), and most of these patients had chronic atrial fibrillation (28 of 30, 93%). Only 16 patients (16 of 30, 53%) were found to have LA thrombi by transthoracic echocardiography. Furthermore, our data showed poor correlation between the echogenicity of LA thrombi and the degree of thrombus organization. Thus, TEE is excellent for detecting LA thrombi in patients with rheumatic heart disease severe enough to warrant mitral valve operations. PMID- 8249845 TI - Effects of changes in management of active infective endocarditis on outcome in a 25-year period. AB - The clinical outcome and long-term follow-up of 130 consecutive patients (141 episodes) with active infective endocarditis who were treated between 1966 and 1991 were analyzed. There was a shift toward a higher proportion of referred patients (39 to 78%), patients aged > 60 years (11 to 41%) and urgent surgical treatment (11 to 44%). Medical treatment was administered in 98 patients (70%); 30-day mortality was 27%. Surgery was performed in 43 patients (30%), with an operative mortality of 26%; 9 of 14 patients (64%) who underwent operation within the first week of admission died. Patients with severe heart failure are at the highest risk for early mortality (relative risk = 21.1; 95% confidence interval 7.4-60.3). Referred patients were much more often treated surgically than were nonreferred patients (48 versus 14%) and had a lower operative mortality (24 vs 30%). Nonreferred patients were more often treated medically (86 vs 52%) and with lower mortality (19 vs 39%). The total follow-up time was 730 patient-years; only 1 patient was considered lost to follow-up. The overall cumulative 5-year and 10 year survival after hospital discharge for patients after urgent surgery were 84 +/- 7% and 53 +/- 7%, respectively, and for those after medical treatment 84 +/- 5% and 77 +/- 6%, respectively. The probability of remaining free of late events (recurrent endocarditis, late valve replacement or death) during 5 and 10 years for patients after urgent surgery was 84 +/- 7% and 53 +/- 15%, respectively, and for those after medical treatment 59 +/- 6% and 40 +/- 7%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249846 TI - Radiofrequency ablation of accessory pathways associated with congenital heart disease including heterotaxy syndrome. AB - Congenital heart disease complicates the management of most accessory pathway mediated tachycardias and also increases the challenge of radiofrequency ablation. Since 1990, radiofrequency ablation of accessory atrioventricular (AV) pathways has been attempted in 10 patients (age range 3.5 months to 30 years) with congenital heart disease: Ebstein's anomaly (n = 5), heterotaxy with AV discordance (n = 3), tetralogy of Fallot (n = 1) and total anomalous pulmonary venous return (n = 1). Eight patients had manifest Wolff-Parkinson-White syndrome and 2 had concealed pathways. Five patients had multiple pathways including 4 of the 5 with Ebstein's anomaly. Of 16 pathways total, 15 were associated with the tricuspid valve including all pathways in the patients with Ebstein's anomaly and heterotaxy. The His bundle area was identified in all patients and involved an anterior AV node in 2 of 3 with heterotaxy. Ablation was performed on the atrial side of the AV ring in all cases. No instance of AV block was encountered. Complete success was achieved in 6 patients including the 3 with heterotaxy. In 2 patients, manifest preexcitation was eliminated and clinical symptoms were greatly modified. The procedure was transiently successful in 1 patient who later had surgical interruption of the accessory pathway during tetralogy of Fallot repair. Ablation was unsuccessful in 1 patient. Thus, the overall success was 80%. PMID- 8249847 TI - Fate of right ventricular hypertrophy in tetralogy of Fallot after corrective surgery. AB - To elucidate the reversibility of right ventricular (RV) myocardial hypertrophy in tetralogy of Fallot (TF), 30 patients underwent RV endomyocardial biopsies 1 to 25 years (mean 9.7 +/- 6.6) after corrective surgery. Myocardial cell diameter was evaluated by comparing histopathologic data with preoperative patients with TF and normal subjects. As a whole, postoperative cell diameter was smaller than that of age-matched preoperative patients with TF, and larger than that of age matched normal subjects. In 7 patients without significant residual pulmonary stenosis whose preoperative data were also available, cell diameter significantly decreased after surgery (17.1 +/- 2.1 to 14.0 +/- 2.1 microns, p < 0.01). There was a positive correlation between postoperative cell diameter and age at study (p < 0.01). To negate the influence of age, cell diameter was expressed in terms of a percentage of age-matched normal values (percent cell diameter). There was no significant correlation between percent cell diameter and age at surgery, age at study or the follow-up periods. There were positive correlations between percent cell diameter and the following parameters: RV systolic pressure (p < 0.05), percent normal RV end-diastolic (p < 0.05) and end-systolic (p < 0.01) volumes. These results demonstrate that RV myocardial hypertrophy in TF can regress to some extent after corrective surgery if significant residual pulmonary stenosis is avoided. PMID- 8249848 TI - Usefulness of biplane transesophageal echocardiography in neonates, infants and children with congenital heart disease. AB - A study was performed to assess the feasibility, additional diagnostic value and potential applications of biplane transesophageal echocardiography in neonates, infants and children. One hundred thirty-two consecutive studies were attempted in 111 anesthetized children with congenital heart disease. Longitudinal and transverse planes were compared using 3 methods: (1) separate 7 mm longitudinal and transverse pediatric transducers used sequentially; (2) an experimental 9 x 8 mm biplane pediatric transducer; and (3) a standard adult biplane transducer (12 x 9 or 13 x 9 mm). In all but 1 patient, a probe could be inserted. The longitudinal plane provided superior visualization of both the right and left ventricular outflow tracts, the interatrial septum, the main pulmonary artery, the ascending aorta and the right coronary artery. In 18 patients (16%), the longitudinal plane provided completely new diagnostic information that was not obtained with combined transthoracic and transverse plane transesophageal echocardiography. However, the transverse plane was mandatory for demonstration of the 4-chamber view, short-axis cross sections through the great arteries, the distal right pulmonary artery and bifurcation of the left coronary artery. The longitudinal plane is complementary to the transverse plane, but cannot substitute for it. PMID- 8249849 TI - Impact of transesophageal echocardiography on the anticoagulation management of patients admitted with focal cerebral ischemia. AB - Transesophageal echocardiography (TEE) improves the diagnostic accuracy of transthoracic echocardiography in the identification of potential cardiac sources of embolus. However, there are few studies of the impact of TEE on the medical management of patients with focal cerebral ischemia. The records of 52 consecutive, hospitalized patients undergoing both TEE and transthoracic echocardiography for suspected cardiac source of embolus were reviewed to determine the influence of TEE on the decision to anticoagulate patients. Of 52 patients, 39 had focal cerebral ischemia (transient ischemic attack, n = 9; acute cerebral infarction, n = 30). In 4 of these 39 patients (10%), the TEE results changed the management of anticoagulation. In 19 of 39 patients (49%), the TEE results helped confirm anticoagulation decisions, and in 16 (41%), the results had no effect on anticoagulation decisions, because of overriding clinical information. Ten of the latter 16 patients had TEE evidence for a possible source of an embolus, but were not anticoagulated; 5 of these were poor candidates for long-term anticoagulation, and the others had right-to-left shunting across a patent foramen ovale or an interatrial septal aneurysm. Clinical variables (atrial fibrillation, TEE findings and pre-TEE anticoagulation status) were considered as possible predictors of post-TEE anticoagulation status using logistic regression analysis; the strongest predictor of post-TEE anticoagulation status was pre-TEE anticoagulation status (p < 0.0005). Despite the selection of patients presumed to receive maximal benefit from TEE, this study suggests that TEE findings are not predictive of subsequent anticoagulation management. However, TEE is at least confirmatory of anticoagulation decisions in most cases. PMID- 8249850 TI - Hydrogen inhalation for detecting intracardiac left-to-right shunting in adults. AB - The hydrogen inhalation technique is easily performed and exquisitely sensitive for detecting intracardiac left-to-right shunting. Previous studies of this technique relied heavily on data from infants and children, and the distinction between "normal" and "abnormal" was imprecise and somewhat arbitrary. The present study was done to assess the results of hydrogen inhalation in adults with and without intracardiac left-to-right shunting. In 45 adult subjects (15 men, 30 women, aged 18 to 72 years) (18 without and 27 with intracardiac left-to-right shunting), the elapsed time from hydrogen inhalation to its appearance in the pulmonary artery was measured. The 18 patients without shunting had an appearance time of 12 +/- 3 (mean +/- SD) seconds, and it was > or = 9 seconds in all. In contrast, the 27 subjects with shunting had an appearance time of 1.5 +/- 0.7 second, with only 1 of 27 being > or = 3 seconds. There was no relation between the magnitude of left-to-right shunting and the hydrogen appearance time. Thus, the hydrogen inhalation technique easily and reliably separates adult subjects with and without intracardiac left-to-right shunting, with no overlap between the 2 groups. PMID- 8249851 TI - Comparison of the Asymptomatic Cardiac Ischemia Pilot and modified Asymptomatic Cardiac Ischemia Pilot versus Bruce and Cornell exercise protocols. AB - The Asymptomatic Cardiac Ischemia Pilot (ACIP) and modified ACIP treadmill exercise protocols were developed to test patients with coronary artery disease and to linearly increase work load between stages. The physiologic changes that occurred with ACIP and modified ACIP were compared to those with the Bruce and Cornell protocols in 28 normal subjects and 16 men with coronary artery disease. The exercise protocols were randomly assigned over 2 days, and gas exchange data were obtained continuously with each test. In normal subjects, the peak heart rate, systolic blood pressure, peak oxygen consumption rate (VO2) and minute ventilation were similar for the 4 protocols tested, with exercise time shortest for the Bruce protocol in comparison with the ACIP, modified ACIP and Cornell protocols (10.2 +/- 3.1 vs 13.4 +/- 4.9, 13.9 +/- 4.5, and 15.0 +/- 4.2 minutes, respectively; p < 0.001). The difference between predicted and observed VO2 was smallest for the ACIP protocol (37.0 +/- 11.0 vs 35.8 +/- 13.5 ml/kg/min) and greatest for the Bruce protocol (41.1 +/- 11.8 vs 36.7 +/- 15.0 ml/kg/min) in normal subjects, as well as in patients with coronary artery disease (ACIP protocol 26.9 +/- 7.1 vs 22.5 +/- 6.7, and Bruce protocol 29.1 +/- 7 vs 22.6 +/- 5.7 ml/kg/min, respectively). The ratio of VO2 to work rate, expressed as a slope, was similar in normal subjects for the 4 protocols tested. However, in patients with coronary artery disease, the slope was 0.84 and 0.83 for the ACIP and modified ACIP protocols, respectively, versus 0.61 and 0.71 for the Bruce and Cornell protocols, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8249852 TI - On-line estimation of changes in left ventricular stroke volume by transesophageal echocardiographic automated border detection in patients undergoing coronary artery bypass grafting. AB - Echocardiographic automated border detection can determine the interface between blood and myocardial tissue and calculate left ventricular (LV) cavity area in real-time. The objective was to determine if on-line measurements of LV cavity area by transesophageal automated border detection could be used to determine beat-to-beat changes in stroke volume in humans. Studies were attempted on 9 consecutive patients, aged 66 +/- 8 years, undergoing coronary bypass surgery. Stroke volume was measured by electromagnetic flow from the ascending aorta, and LV cavity area was measured at the midventricular short-axis level. Simultaneous area and flow data were recorded on a computer workstation through a customized interface with the ultrasound system. Recordings were performed during baseline apnea and rapid alterations induced by inferior vena caval occlusions before and after cardiopulmonary bypass. Measurements of stroke area (maximal area-minimal area) were correlated with stroke volume for matched beats. Data were available for analysis on 8 of 9 patients before and on 5 patients after cardiopulmonary bypass for 644 beats. Stroke area was closely correlated with stroke volume both before (mean R = 0.94 +/- 0.03, SEE = 0.33 +/- 0.12 cm2) and after (mean R = 0.92 +/- 0.05, SEE = 0.59 +/- 0.81 cm2) cardiopulmonary bypass. The slopes of these stroke area-stroke volume relations were quite reproducible from before to after cardiopulmonary bypass in the same patient but varied between individual patients. Transesophageal automated border detection has potential for on-line estimation of changes in stroke volume in selected patients. PMID- 8249853 TI - Technetium-99m teboroxime regional myocardial washout in subjects with and without coronary artery disease. AB - The aim of this study was to test the hypothesis that regional myocardial washout of technetium-99m teboroxime is slowed in the presence of coronary stenosis. Washout was assessed in 33 catheterized patients and in 13 with a low likelihood of coronary artery disease, using a triple detector camera and dynamic single photon emission computed tomography, with serial 1-minute acquisitions after injection of 20 to 25 mCi of teboroxime at the third minute of adenosine-induced hyperemia. Washout was measured as the percent change in counts between the first, second and third minutes after injection, as measured in 6 short-axis myocardial regions of interest. Myocardial regions were classified as ischemic (> or = 50% diameter stenosis and no prior myocardial infarct), infarcted, normal (no significant coronary stenosis) or "low likelihood" (from the 13 patients with a low likelihood of coronary artery disease). Teboroxime washout was significantly (p < 0.001) slowed in the ischemic myocardium (12.7 +/- 8.3%) compared with the normal (18.5 +/- 5.7%), low-likelihood (17.8 +/- 6.1%) and infarcted (17.8 +/- 4.4%) zones. There was regional variability in washout rates (% washout/min), with the anterior wall having the lowest (13.8 +/- 3.4%/min) and the inferior wall the highest (20.7 +/- 7.9%/min) values. In regard to individual coronary territories, 21 of 41 ischemic, noninfarcted territories (51%) had abnormal washout compared with 3 of 43 normal territories (7%) (p = 0.001). In conclusion, regional washout of teboroxime is detectably slowed in ischemic, noninfarcted myocardium. The clinical value of washout analysis in teboroxime single-photon emission computed tomography warrants further investigation. PMID- 8249854 TI - Radionuclide angiography for risk stratification of patients with coronary artery disease. AB - Radionuclide angiography has become an accepted diagnostic method that is relied upon for its rapid, accurate and quantitative capabilities to characterize LV function. This method will continue to be used extensively in the future for the evaluation of patients with CAD. In addition to its common use to measure LV function at rest, the application of radionuclide angiography to assess LV function during exercise has proven prognostic potential that can be used in the clinical decision-making process in patients with known or suspected CAD. PMID- 8249855 TI - Little women, Texas, small numbers and coronary artery bypass grafting. PMID- 8249856 TI - Familiar clustering of aortic dissection in polycystic kidney disease. PMID- 8249857 TI - Massive diltiazem overdose. PMID- 8249858 TI - Getting cardiologists interested in lipids. PMID- 8249859 TI - Pulsatile varicose veins in tricuspid regurgitation. PMID- 8249860 TI - Radiation exposure from percutaneous transluminal coronary angioplasty. PMID- 8249861 TI - Nutrition education in medical schools. PMID- 8249862 TI - Isn't it time to teach nutrition to medical students? PMID- 8249863 TI - Adipose tissue distribution as assessed by magnetic resonance imaging and total body fat by magnetic resonance imaging, underwater weighing, and body-water dilution in healthy women. AB - A method for estimating total body fat (TBF) and the amount and distribution of adipose tissue in humans by using magnetic resonance imaging (MRI) is described. Thirty transaxial images of subjects were taken by a nuclear magnetic imager (0.02 T). Adipose tissue volume was calculated from adipose tissue area in each image and from the distances between images. The CV when estimating total volume of body adipose tissue was 1.5%. TBF of healthy Swedish women was estimated by the MRI method (n = 25), by underwater weighing (UWW) (n = 20), and by body-water dilution (BWD) (n = 23). When assessed by MRI and BWD, women contained 1.4 +/- 2.9% less and 4.7 +/- 4.0% more TBF, respectively, than when assessed by UWW. On average, 74.2% of total adipose tissue volume was situated subcutaneously. Subcutaneous fat:fat-free body weight and nonsubcutaneous fat:fat-free body weight were both significantly correlated with percent TBF but the women stored excess fat subcutaneously rather than nonsubcutaneously. PMID- 8249864 TI - Validation and application of dual-energy x-ray absorptiometry to measure bone mass and body composition in small infants. AB - Precision and validity of dual-energy x-ray absorptiometry (DXA) for analysis of whole-body composition in infants were assessed by 1) scanning piglets in triplicate to calculate CVs, and 2) comparing DXA estimates with chemical analysis of whole carcass. The mean CVs for all DXA measures in small piglets and large piglets were < 2.5%, except for fat mass, which were 6.3% and 3.5%, respectively. In large piglets DXA provided reasonable estimates of chemical analysis for bone mineral content (BMC), lean body mass, and fat mass, but only for lean body mass in small piglets. DXA overestimated fat by twofold and underestimated BMC by a third in small piglets. Scans of prematurely born infants (n = 17) at term and at 3, 6, and 12 mo corrected age demonstrated that changes in BMC, lean body mass, and fat mass can be quantitated by DXA. However, further refinement of DXA technology is necessary before reliable measures of BMC and fat mass in small infants are attainable. PMID- 8249865 TI - Dietary fat affects weight loss and adiposity during energy restriction in rats. AB - This study examined the effect of dietary fat during food restriction on weight loss, body composition, and adipose tissue lipoprotein lipase activity (ATLPL). Rats previously fed a high-fat diet were divided into four groups. One group served as baseline controls. The other three groups were fed diets of 12%, 28%, or 45% fat, restricted (R) in amounts to equal 75% of their previous ad libitum energy intake. After 10 wk, body weight loss and reduction of adiposity were significantly less in the 45%-R group compared with the 12%-R group (weight P < 0.05; adiposity P < 0.01). Rats fed restricted amounts of the high-fat diet (45% R group) retained adiposity equivalent to baseline control rats. ATLPL activity was increased in the 45%-R group compared with baseline control rats. Further studies are needed in humans, but these results suggest that body composition will not be normalized with reduction of energy intake unless dietary fat is also reduced. PMID- 8249866 TI - Effect of a weight cycle on visceral fat accumulation. AB - Magnetic resonance imaging was used to study the effect of a single weight cycle on three fat depots: the visceral and subcutaneous abdominal depots and the subcutaneous depot at the trochanter level. Obese subjects (17 men, 15 women) were examined before and after weight-loss intervention and 67 wk after intervention. They lost 12.9 +/- 3.3 (mean +/- SD) kg body wt during intervention and regained 11.9 +/- 5.1 kg during follow-up. Weight regain did not result in greater body fatness than before weight loss (initial fat mass: 34.3 +/- 6.1 kg, final fat mass: 32.8 +/- 7.7 kg; P = 0.047). There was no indication of a preferential deposition of visceral fat after weight regain (initial visceral fat area: 120 +/- 41 cm2, final visceral fat area: 110 +/- 48 cm2; P = 0.087). On the contrary, there was a slight tendency to accumulate subcutaneous fat at the expense of visceral fat. It is concluded that weight loss followed by weight regain neither leads to a greater body fatness nor to a larger amount of visceral fat compared with before weight loss. PMID- 8249867 TI - Nutritional factors and anticonvulsant therapies: effect on growth in children with epilepsy. AB - Factors influencing growth before puberty were examined in a group of 32 epileptic children in a cross-sectional investigation. Participants in the study were divided into two groups according to what anticonvulsant drug, carbamazepine or valproic acid, they were currently taking. Dietary intakes of energy, iron, and zinc were assessed and hemoglobin and serum zinc concentrations were evaluated. Measurements of height and weight were compared with standard growth charts prepared by the National Center for Health Statistics, Hyattsville, MD. A Student's t test comparison was made along with analyses of covariance and stepwise regression, and no significant differences were found between the two groups of children in terms of height or weight. Linear growth was normal. We found that low dietary zinc intake, 64-87% of the recommended dietary allowance, was not associated with reduced height in children. In addition, neither the number of years taking anticonvulsant medication nor the total amount of drug intake had a significant effect on weight. This was an unanticipated result because valproic acid has been associated with weight gain because of increased appetite. PMID- 8249868 TI - A positive association between extended breast-feeding and nutritional status in rural Hubei Province, People's Republic of China. AB - Data were analyzed from a cross-sectional nutrition surveillance survey to determine the association between extended breast-feeding and growth. The sample consisted of 2148 initially breast-fed children between 12 and 47 mo of age. Breast-feeding for > 24 mo was associated with a greater height-for-age Z score, and breast-feeding for > 18 mo was associated with greater weight-for-age and weight-for-height Z scores. These results remained significant after the number of food groups being consumed at 12 mo of age, age when the selected food items were first given to a child, the consumption of powered milk, recent infections, age, sex, birth order, birth weight, county of residence, father's occupation, and mother's education were controlled for. These results suggest that extended breast-feeding in this population, in which food was introduced late in infancy, was associated with improved nutritional status as measured by standard anthropometric indicators. PMID- 8249869 TI - Effect of dietary macronutrient content on carnitine excretion and efficiency of carnitine reabsorption. AB - We examined the effect of macronutrient content on glomerular filtration rate (GFR), and excretion, reabsorption, and filtered load of carnitine. Ten subjects consumed five diets [high protein (HP), low protein (LP), control, high fat (HF), and high carbohydrate (HC)] of equal energy and carnitine content for 6 d each, in a randomized crossover manner. The rate of carnitine excretion was lower after the LP diet than after the HP diet because of lower GFR after the LP diet. The rate of carnitine reabsorption was lower after the LP diet than after the HP diet, also because of the lower GFR after the LP diet. The rate of carnitine reabsorption was not different after the HF and HC diets, nor was GFR. The filtered load of carnitine, however, was greater after the HF diet, resulting in a higher rate of carnitine excretion. PMID- 8249870 TI - Prediction of glycemic index for starchy foods. AB - Associations between published values for glycemic index (GI) and chemical components were examined by regression analysis in 18 starchy foods. Independent variables were the amounts of protein, fat, phytic acid, and total dietary fiber (TDF) present in food portions containing 50 g available carbohydrate. GI was similarly and significantly correlated (P < 0.05) with TDF, protein, and phytate. When foods were considered as comprising two groups, legumes and others (grains and tubers), food category accounted for as large a fraction of variability in GI as did regression analysis with food components in the undivided sample. Most correlations between GI and components reverted toward zero when calculated only for the legumes or the "others" group. Evidence is cited that the method of preparing foods and the characteristics of starch and starch granules are more important in predicting GI among starchy food than is the content of any of these four food components. PMID- 8249871 TI - Adaptation of lactose maldigesters to continued milk intakes. AB - Twenty-five lactose-maldigesting and lactose-intolerant African Americans, ranging in age from 13 to 39 y, were given gradually increasing amounts of lactose in milk over a period of time until the maximum lactose dose tolerated was determined. Seventeen (77%) of the 22 subjects who completed the study tolerated > or = 12 g lactose and 5 (23%) tolerated < 12 g. Breath-hydrogen tests done on each subject with the maximum dose of lactose tolerated showed that only four (18%) had a breath-hydrogen concentration < 5 ppm above fasting concentration. This study suggests that the majority of African-American young adults who claim intolerance to moderate amounts of milk can ultimately adapt and tolerate > or = 12 g lactose in milk (the equivalent of 8 oz of full-lactose milk) with minimal or no discomfort if milk is ingested in gradually increasing amounts. The mechanism of adaptation is assumed to be an increased tolerance to colonic lactose-fermentation products. PMID- 8249872 TI - Effects of skin thickness, age, body fat, and sunlight on serum 25-hydroxyvitamin D. AB - We tested the hypothesis that the age-related decline in skin thickness may contribute to the age-related decline in serum 25-hydroxyvitamin D [25(OH)D]. We measured skinfold thickness on the back of the hand, serum 25(OH)D, height, and weight in 433 normal postmenopausal women. We also noted the average daily hours of sunlight in the month in which the observations were made and in the preceding 2 mo. Serum 25(OH)D was positively related to hours of sunlight (with a time lag of 2 mo) and to skin thickness, and negatively to body mass index (wt/ht2). Serum 25(OH)D fell significantly after age 69 y. Seasonal variation of serum 25(OH)D was greater in lean than in fat subjects, which we attributed to the larger fat mass and consequent larger pool size in the latter group. The results suggest that the tendency for serum 25(OH)D to fall with age is due in part to the age related decline in skin thickness. PMID- 8249873 TI - Reduced plasma ascorbic acid concentrations in nonsmokers regularly exposed to environmental tobacco smoke. AB - Oxidants in cigarette smoke accelerate metabolic turnover of ascorbic acid (AA), and thereby deplete body stores of this potent antioxidant and putative anticarcinogen in active smokers. We examined plasma AA concentrations and vitamin C intakes in nonsmokers regularly exposed to environmental tobacco smoke (ie, passive smokers; n = 44), as compared with active smokers (n = 47) and nonexposed nonsmokers (n = 50), to determine whether passive smokers also exhibit altered AA nutriture suggestive of oxidant exposure. Plasma AA concentrations in passive smokers were intermediate between those of active smokers (P = 0.0001) and nonexposed nonsmokers (P = 0.01) despite similar dietary vitamin C intakes. Hypovitaminosis C (< 23 mumol/L) was observed in 24% of active smokers and 12% of passive smokers but not in nonexposed nonsmokers. Reduced plasma AA concentrations were associated with low vitamin C intakes within smoke-exposed populations only. We conclude that chronic smoke exposure, particularly in association with low vitamin C intake, may reduce AA pools in both active and passive smokers. PMID- 8249874 TI - Amelioration of effects of severe dietary copper deficiency by food restriction in rats. AB - To test the effect of food restriction on responses to dietary copper deficiency, male weanling rats were fed two amounts of dietary copper via five dietary-intake regimens ranging from ad libitum to 70% of ad libitum intake. Copper-deficient rats exhibited characteristic signs, including depressed organ copper content, reduced activity of copper-dependent enzymes, enlarged hearts, and anemia. Food restriction attenuated the cardiac enlargement, red blood cell defects, and reduction of superoxide dismutase activity in copper-deficient rats. Mineral and enzyme assays suggested that possible mechanisms for this amelioration are the correction of copper status and/or the improvement of antioxidant status. Also, food restriction depressed serum cholesterol and enhanced cytochrome c oxidase activity in both copper-adequate and copper-deficient rats, which compensated for effects of copper deficiency. A second experiment illustrated that the mortality associated with severe copper deficiency was also inhibited by food restriction. PMID- 8249875 TI - Zinc and copper balances in healthy adult males during and after 17 wk of bed rest. AB - The effects of long-term bed rest on zinc and copper balances were measured in seven healthy men. Volunteers aged 22-54 y (mean +/- SD, 34 +/- 12 y), 168-185 cm in height (173 +/- 5 cm), and 64-86 kg in weight (74 +/- 9 kg) remained on a metabolic ward for 29 wk. Subjects were ambulatory during weeks 1-5, remained in continuous bed rest for weeks 6-22, and were reambulated during weeks 23-29. Copper and zinc were measured in weekly urine and fecal composites. Dietary intakes provided (mean +/- SD) 19.2 +/- 1.2 mumol Cu (1.22 +/- 0.08 mg), 211 +/- 11 mumol Zn (13.81 +/- 0.72 mg), 25.2 +/- 1.2 mmol Ca (1011 +/- 46 mg), 1086 +/- 46 mmol N (15.21 +/- 0.65 g), and 48.1 +/- 1.4 mmol K (1489 +/- 44 mg)/d. Bed rest increased fecal zinc excretion and decreased zinc balance, whereas copper balance was unchanged. Reambulation decreased fecal zinc excretion and increased both zinc and copper balances. These results suggest that during long-term bed rest or space flight, individuals will lose total body zinc and will retain more zinc and copper when they reambulate. PMID- 8249876 TI - Measurement of zinc bioavailability from beef and a ready-to-eat high-fiber breakfast cereal in humans: application of a whole-gut lavage technique. AB - We measured zinc absorption in humans from two different foods, using a whole-gut lavage technique. Healthy adults (n = 8) were given test foods containing approximately 4 mg (61.2 mumol) Zn as either beef or a high-fiber, ready-to-eat breakfast cereal. Basal zinc loss in the gut was measured after giving only 200 mL water alone after an overnight fast. Mean polyethylene glycol (PEG) recovery, a nonabsorbable fecal marker, was 95 +/- 2% (mean +/- SE). There was no significant difference of PEG recoveries with the various treatments. True absorption of zinc was 11.0 +/- 5.4 mumol (716.3 +/- 355.9 micrograms) for breakfast cereal and 34.8 +/- 3.5 mumol (2274.1 +/- 230.2 micrograms) for beef. Fractional zinc absorption was significantly lower for breakfast cereal compared with beef, 14.7 +/- 6.7% vs 55.1 +/- 4.6%, respectively. Our findings demonstrate that zinc absorption can be measured in humans from single foods containing a physiological dose of zinc by using the whole-gut lavage absorption technique and that zinc bioavailability from beef is about fourfold greater than from a high fiber breakfast cereal. PMID- 8249877 TI - Metabolism of triacetin-derived acetate in dogs. AB - Triacetin is a water-soluble triglyceride that may have a role as a parenteral nutrient. In the present study triacetin was administered intravenously to mongrel dogs (n = 10) 2 wk after surgical placement of blood-sampling catheters in the aorta and in the portal, hepatic, renal, and femoral veins. [1-14C]Acetate was infused to allow quantification of organ uptake of acetate as well as systemic turnover and oxidation. Systemic acetate turnover accounted for approximately 70% of triacetin-derived acetate, assuming complete hydrolysis of the triglyceride. Approximately 80% of systemic acetate uptake was rapidly oxidized. Significant acetate uptake was demonstrated in all tissues (liver, 559 +/- 68; intestine, 342 +/- 23; hindlimb, 89 +/- 7; and kidney, 330 +/- 37 mumol/min). In conclusion, during intravenous administration in dogs, the majority of infused triacetin undergoes intravascular hydrolysis, and the majority of the resulting acetate is oxidized. Thus, energy in the form of short chain fatty acids can be delivered to a resting gut via intravenous infusion of a short-chain triglyceride. PMID- 8249878 TI - Effect of parenteral administration of short-chain triglycerides on leucine metabolism. AB - The present studies investigated the effects of intravenous administration of the short-chain triglyceride triacetin on leucine metabolism in dogs. Animals received infusions of triacetin at 1.0 x estimated resting energy expenditure (REE), hyperenergetic triacetin at 1.5 x REE, glycerol, or saline during infusion of [1-14C]leucine. During both triacetin infusions, plasma alpha-ketoisocaproate concentrations increased (P < 0.05). During triacetin infusion at 1.5 REE, the plasma leucine concentration decreased (P < 0.05) and leucine rate of appearance decreased by approximately 19% (P < 0.05); this was significantly greater than the changes that occurred during triacetin at 1.0 x REE and glycerol (P < 0.05). There was no difference in leucine oxidation between the dogs given triacetin at 1.0 x REE and control groups, whereas leucine oxidation decreased by 53% during triacetin infusion at 1.5 x REE (P < 0.05). Nonoxidative leucine disappearance, an indicator of protein synthesis, did not change in any of the studies. These results indicate that triacetin has effects on leucine metabolism similar to those previously reported with long-chain triglyceride emulsions. Because of its water solubility, lack of toxicity, and favorable effects on protein metabolism, further studies are warranted regarding the use of triacetin as a parenteral nutrient. PMID- 8249879 TI - Bile acids and pH values in total feces and in fecal water from habitually omnivorous and vegetarian subjects. AB - Twenty habitually omnivorous subjects and 19 habitually lactoovovegetarian subjects aged 59-65 y collected feces during 4 consecutive days. The concentrations of bile acids in total feces did not differ between the omnivores and vegetarians, but the bile acid concentrations in fecal water were significantly lower in the vegetarians. The concentration of the colorectal cancer-predicting bile acid deoxycholic acid in fecal water was explained by the intake of saturated fat and the daily fecal wet weight (r2 = 0.50). Fecal pH did not differ between the omnivores and vegetarians. This variable was significantly (P < 0.05) explained by the intake of calcium (r2 = 0.30); 24-h fecal wet weight and defecation frequency were significantly higher in the vegetarians. In conclusion, our vegetarian subjects had a lower concentration of deoxycholic acid in fecal water, higher fecal wet weight, and higher defecation frequency than the omnivorous subjects. PMID- 8249880 TI - Maintenance of iron status in healthy men during an extended period of stress and physical activity. AB - Body weight loss and iron status of 55 male soldiers were measured during 62 d of intense physical activity and sustained stress and sleep deprivation. Body weight declined from (mean +/- SD) 75.9 +/- 9.0 to 63.8 +/- 6.7 kg (P < 0.05). Serum iron fell from 13.7 +/- 5.6 to 8.3 +/- 3.6 mumol/L by 14 d (P < 0.05), but returned to baseline values by 8 wk. Total iron-binding capacity declined from 53.4 +/- 6.8 at baseline to 47.5 +/- 6.3 mumol/L at 8 wk (P < 0.05). During the study, hematocrit, serum hemoglobin, and erythrocyte count did not change, whereas ferritin increased from 116 +/- 84 to 202 +/- 106 micrograms/L (P > 0.05). Adequate dietary iron, initiation of training with adequate body iron stores, and physical activity not in excess of pretraining workloads contributed to the maintenance of iron status during prolonged physical activity and stress. Our results suggest that some acute phase-like disturbances in iron metabolism may be a normal component of adaptation to stress and physical activity in healthy men. PMID- 8249881 TI - The role of Harvard's School of Public Health in development of nutrition education for medicine. PMID- 8249882 TI - The Friedewald equation for the determination of low-density-lipoprotein cholesterol: a special case. PMID- 8249883 TI - Medical complications of cocaine abuse. PMID- 8249884 TI - The antiphospholipid-thrombosis syndromes. Fact, fiction, confusion, and controversy. AB - Anticardiolipin antibodies and the lupus anticoagulant are strongly associated with thrombosis and appear to be the most common of the acquired blood protein defects causing thrombosis. Although the precise mechanism(s) whereby antiphospholipid antibodies alter hemostasis to induce a hypercoagulable state remain unclear, several theories have been advanced. Because the aPTT is unreliable in patients with lupus anticoagulant and is not usually prolonged in patients with anticardiolipin antibodies, definitive tests, such as ELISA for IgG, IgA, and IgM anticardiolipin antibodies and the dRVVT (followed by cephalin correction for confirmation) for lupus anticoagulant, should be immediately ordered when suspecting antiphospholipid syndrome in persons with otherwise unexplained thrombotic or thromboembolic events or fetal wastage syndrome. The laboratory diagnosis of APL-T syndrome is summarized in Figure 1. PMID- 8249885 TI - Blackened bronchoalveolar lavage fluid in crack smokers. A preliminary study. AB - A retrospective study was performed on heavily pigmented pulmonary cytologic specimens from 14 hospital patients to determine the clinical features distinguishing these cases. The lavage fluid or sputum in each case was turbid and gray or black, exceeding the blackness usually seen in heavy tobacco smokers dwelling in the same urban environment. Excessive carbonaceous material was observed in the cytoplasm of pulmonary alveolar macrophages or the extracellular compartment of the smears. The latter feature is not seen in cigarette smokers. Many other pigmentary sources were ruled out, including melanin, hemosiderin, medicinal charcoal, India ink, and hematoxylin crystals. The common feature of the patients was that they recently or currently smoked the crack form of cocaine heavily; five patients also had positive toxicologic results for cocaine at admission. The authors suggest that blackened bronchoalveolar lavage fluid indicates the possibility of crack cocaine smoking and the associated sequelae, particularly when the carbonaceous material is present in the extracellular compartment. PMID- 8249886 TI - Calcium oxalate is associated with benign breast tissue. Can we avoid biopsy? AB - Breast biopsies are commonly performed for abnormal, usually clustered, calcifications detected by mammography. Calcium phosphate is the predominant form of calcium seen in breast tissue and is frequently associated with malignancy. Calcium oxalate, which can also be present in breast tissue, has been exclusively associated with benign lesions. Thus, if mammography could distinguish calcium phosphate from calcium oxalate, biopsy could be avoided in some patients. Pathologic findings and corresponding mammograms of 55 patients who underwent biopsy for abnormal calcifications were reviewed. The authors evaluated such pathologic features as type of calcification, anatomic location, and association with fibrocystic changes or carcinoma. Mammographically, calcifications were categorized by size, distribution, and morphology, and each was assigned a density rating of low, medium, or high. Of the 55 cases, 41 contained calcium phosphate only, 8 contained calcium oxalate only, and 6 contained both. If only calcium oxalate was present, the calcium was always associated with benign epithelium. Of 47 cases, calcium phosphate was associated with benign breast disease in 28 and with carcinoma in 19. Five of six cases with both calcium phosphate and calcium oxalate contained carcinoma; calcium phosphate was seen in the carcinoma area in all five. Radiologically, calcium phosphate was typically medium to high density, whereas calcium oxalate was characterized as amorphous, low to medium density. Other low-density calcifications were almost always benign, unless pleomorphic in shape. Although further work is necessary to confirm these findings, it appears that, radiologically, low-density, amorphous, calcifications, even if clustered, are associated with benign breast disease, and may represent calcium oxalate. Patients with such calcifications may be managed conservatively. PMID- 8249887 TI - Muscle involvement in Walker-Warburg syndrome. Clinicopathologic features of four cases. AB - Walker-Warburg syndrome is a lethal autosomal recessive disorder characterized by anomalies of the central nervous system and the eye. Recently, elevation of muscle enzymes and congenital muscular dystrophy have been found to contribute to the diagnosis of this syndrome. The authors studied two pairs of siblings with Walker-Warburg syndrome, offspring of consanguineous parents from two unrelated families. The patients had hydrocephalus, congenital glaucoma, elevated muscle enzymes, and hypotonicity. The histologic and ultrastructural muscular changes were consistent with congenital muscular dystrophy. PMID- 8249888 TI - Use of cell image analysis in the detection of cancer from specimens obtained during endoscopic retrograde cholangiopancreatography. AB - The authors report a scoring system based on cell image analysis of specimens obtained during endoscopic retrograde cholangiopancreatography from either the biliary or pancreatic duct epithelium. Nine normal, 20 inflammatory, and 19 malignant epithelia were assessed using a cell-image processor that permits determination of morphometric (nuclear area), densitometric (nuclear DNA content), and textural (chromatin distribution) parameters on Feulgen-stained nuclei. The score is based on three densitometric parameters: the DNA histogram type, the nuclear DNA content, and the percent proliferating cell population. These parameters permit distinction of normal and inflammatory from malignant epithelium with 95% sensitivity and 100% specificity. Cell-image analysis of specimens obtained during endoscopic retrograde cholangiopancreatography is a useful clinical tool in the detection of malignant lesions of the pancreas and biliary epithelia. PMID- 8249889 TI - Latent sites of Epstein-Barr virus infection. AB - Epstein-Barr virus (EBV) has been detected in a wide spectrum of tumors. This study investigates the detection rate of EBV-DNA by Southern blot hybridization analysis (SOBH) and polymerase chain reaction (PCR) in different tissues from persons without apparent EBV-related diseases. Of 20 tonsillectomy specimens studied, SOBH indicated positivity for EBV-DNA in 1 case, and PCR indicated positivity in 10. In autopsies performed on patients with no apparent evidence of EBV-related diseases, the viral DNA was only detected by PCR in the following: parotid gland (7/15), submandibular gland (8/20), nasopharynx (8/10), tonsil (8/10), larynx (5/6), lung (5/9), cervical lymph node (7/10), mediastinal lymph node (7/10), abdominal lymph node (4/10), spleen (6/10), thyroid (5/10), liver (1/10), pancreas (1/4), kidney (4/10), uterine cervix (1/4), ovary (1/5) and testis (1/3). These results provide a baseline for interpreting the role of EBV in carcinogenesis. PMID- 8249890 TI - Epithelioid angiosarcoma arising in uterine leiomyomas. AB - Primary angiosarcomas of the uterus are exceedingly rare. The authors report a unique case of this entity arising in connection with uterine leiomyomas. The malignant tumor cells showed focally epithelioid features and were immunoreactive for keratin. Electron microscopy and immunohistochemistry confirmed their endothelial nature. PMID- 8249891 TI - High-grade transformation of chronic lymphocytic leukemia and low-grade non Hodgkin's lymphoma. Genotypic confirmation of clonal identity. AB - The abrupt appearance of a high-grade tumor in patients with low-grade malignant lymphoma usually is associated with an accelerated clinical disease course. The high-grade lymphoma may take a variety of histologic forms and often, but not always, represents evolution of the original low-grade disease, as shown by immunophenotypic or immunogenotypic studies. The authors describe the transformation of a variety of low-grade B-cell neoplasms to high-grade tumors in four patients. The initial diagnoses included chronic lymphocytic leukemia and mantle cell lymphoma in one patient each and low-grade follicular lymphoma in two patients. The high-grade tumors were classified as lymphoblastic lymphoma in one patient and small noncleaved cell lymphoma in two patients. The high-grade component manifests primarily in the peripheral blood as circulating blast-like cells consistent with large-cell lymphoma in the remaining patient. In each case, immunophenotypic studies showed identical monoclonal surface immunoglobulin expression on the low- and high-grade tumors. Immunoglobulin heavy chain gene and kappa light chain gene studies showed identical clonally rearranged bands in paired samples from three of the four patients, a finding indicative of clonal identity. Unexpectedly, dissimilar immunoglobulin light and heavy chain gene rearrangements were detected in the paired samples from one patient with previously diagnosed follicular lymphoma, making the relationship of the two tumors from this patient uncertain; however, additional Southern blot analysis of the bcl-2 gene showed identical rearrangements in both lesions. Furthermore, polymerase chain reaction across the t(14;18) major breakpoint region in both tumors amplified nucleotide fragments of identical size, confirming the clonal identity of the low- and high-grade lymphomas despite the divergent immunoglobulin gene studies. These studies show that low-grade malignant lymphomas of small lymphocytic, mantle cell, or follicular small cleaved cell types may assume high-grade morphologic characteristics, that this change is the result of transformation of the preexisting low-grade malignant neoplasm, and that this progression, like typical Richter's syndrome, is associated with a dramatically accelerated clinical course. In addition, these studies confirm previous reports that disparate immunoglobulin light and heavy chain gene rearrangements are not necessarily an indicator of different cellular origins, and additional genotypic studies occasionally may be required to show the clonal identity of the cell population involved in these morphologic transformations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8249892 TI - Sensitive detection of clonal antigen receptor gene rearrangements in non Hodgkin's malignant lymphoma with an anchored polymerase chain reaction-based strategy. AB - The identification of minimal residual disease (MRD) in non-Hodgkin's malignant lymphoma is of immediate clinical relevance. Although several polymerase chain reaction (PCR)-based strategies are currently available, they all share serious technical or theoretic limitations with respect to their ability to detect the broad spectrum of mature B-cell neoplasia. We report a new strategy based on anchored PCR that does not rely on a particular chromosomal translocation or VH consensus sequence. It can yield the complete DNA sequence of the immunoglobulin heavy chain variable region (VHDJH). The assay is capable of detecting one neoplastic cell among 10(3)-10(4) normal cells with high specificity and has the potential to greatly expand the variety of B-cell malignant neoplasms that can be studied. PMID- 8249893 TI - Immunophenotyping of acute leukemia by flow cytometric analysis. Use of CD45 and right-angle light scatter to gate on leukemic blasts in three-color analysis. AB - This article describes a procedure for performing routine three-color flow cytometric analysis for acute leukemia on lysed whole bone marrow preparations. This technique uses the combination of CD45 intensity and right-angle light scatter (RALS) to distinguish leukemic cells from normal lymphocytes, monocytes, neutrophils, eosinophils, and nucleated red blood cells. On this display, leukemic cells occupy a unique blast region characterized by intermediate CD45 density and low RALS, which, in normal marrows, contains less than 5% of the total cells. This approach was applied to 39 cases of acute leukemia and 8 cases of myelodysplasia or myeloproliferative disorders. The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemic cell counts over a wide range. Moreover, the pattern seen on the CD45-RALS display was different for different French-American-British subtypes of leukemia, suggesting that this pattern might be useful for categorization. When CD45-peridin chlorophyll alpha protein was combined with other pairs of fluorescein isothiocyanate- and phycoerythrin-conjugated reagents, it was possible to set an analysis window on the leukemic blasts and display dual-parameter (ie, green vs. red fluorescence) data regarding expression of two additional markers on the leukemic population. This gating strategy was superior to traditional forward angle versus RALS displays in that it did a better job of isolating the leukemic cells analytically. PMID- 8249894 TI - Type 2 Epstein-Barr virus genome and latent membrane protein-1 expression in a T cell-rich lymphoma of probable B-cell lineage. AB - In a 79-year-old white woman, a lymphoproliferative disorder that was associated with type 2 Epstein-Barr virus (EBV) infection or reactivation, documented by three subsequent lymph node biopsies, was studied. After an initial phase with features of reactive lymphadenopathy with exhaustion of the follicular germinal centers and depletion of the B-cell lymphoid population, the disease evolved to a T-cell-rich lymphoma in which a clonal cell population of probable B-cell origin was identified. Such clonal cell population harbored the viral genome and expressed EBV latent membrane protein-1 but not EBV nuclear antigen-2. The implications of immunologic interactions between the clonal EBV-infected cells and the reactive T-cell component in the pathogenetic process are discussed. PMID- 8249895 TI - Immunoglobulin clonality analysis. Resolution of ambiguities in immunofixation electrophoresis results by high-resolution, two-dimensional electrophoretic analysis of paraprotein bands eluted from agarose gels. AB - This article describes evaluation of the clonality of origin of immunoglobulin (Ig) chains by strategies that include qualitative analysis of paraproteins with high-resolution, two-dimensional electrophoresis (2DE) and silver staining. This approach is helpful in evaluating specimens in which standard immunoelectrophoresis or immunofixation electrophoresis (IFE) techniques do not provide definitive Ig typing results. One method the authors developed involves a "band elution" procedure, in which proteins present in high-resolution agarose gel electrophoresis bands are cut from the agarose gel, eluted with a denaturing buffer, and subjected to 2DE. The microheterogeneity patterns of the Ig light chains, heavy chains, or both are evaluated for their relationship regarding mass, charge, and, by inference, number of genes of origin. When necessary, determination of charge relationships may be aided by urea-mediated carbamylation of lysine residues, which introduces single-charge shifts to the individual protein subunits. Overall, these adjunctive techniques are particularly useful in cases with multiple bands of identical immunologic types (eg, several IgG1 bands) on IFE before and after sulfhydryl reduction (with dithiothreitol or 2 mercaptoethanol). The authors present procedural details and examples of the 2DE band elution patterns of serum and urine samples from four patients with B-cell dyscrasias, including the first reported case of POEMS syndrome with biclonal gammopathy, to the best of their knowledge. PMID- 8249896 TI - Lambda III subgroup immunoglobulin light chains are precursor proteins of nodular pulmonary amyloidosis. AB - To investigate the selectivity of amyloid deposits in lungs, the authors determined the amino-terminal sequence of two amyloid proteins of two nodular pulmonary cases--a case of systemic amyloidosis without myeloma and a localized case--and produced a specific antibody to the amyloid protein of the systemic case to stain other amyloid lung specimens. Moreover, the authors extracted immunoglobulins (Igs) from the amyloidotic spleen of the systemic case to assess their affinity with the lung specimens. Both nodular pulmonary amyloid proteins had an amino-terminal structure similar to that of the lambda III subgroup Ig light chain and reacted with the same antibody. In other cases, nodular forms also stained positively with the same antibody, but interstitial forms were unstained. Extracted Igs containing amyloidogenic lambda III light chains reacted with pulmonary amyloid deposits on sections of both nodular cases. These results suggest that the lambda III subgroup Ig light chains are precursor proteins of nodular pulmonary amyloidosis and that their affinity for pulmonary tissue may be a main determinant of pulmonary localization in nodular amyloidosis. PMID- 8249897 TI - ARACHnase. An evaluation of a positive control for platelet neutralization procedure testing with seven commercial activated partial thromboplastin time reagents. AB - ARACHnase (Hemostasis Diagnostics International Co., Denver, CO) is a normal plasma that contains a venom extract from the brown recluse spider, Loxosceles reclusa, which mimics the presence of a lupus anticoagulant (LA). Seven activated partial thromboplastin time (APTT) reagents were used for platelet neutralization procedure (PNP) testing with ARACHnase: Automated APTT (Organon-Teknika, Durham, NC); Thrombofax and Thrombosil (Ortho, Raritan, NJ); Actin and Actin FSL (Dade, Aguado, PR); and Thromboscreen-Kontact and Thromboscreen-APTT LS (Pacific Hemostasis, Ventura, CA). ARACHnase consistently displayed a positive PNP result of greater than 5 seconds correction of the initial baseline APTT. Thus, ARACHnase may provide a positive control for LA testing, regardless of the choice of APTT reagent and activator/phospholipid combination. PMID- 8249899 TI - Reactive histiocytic disorders. PMID- 8249898 TI - Rapid detection of plasma glycocalicin by a latex agglutination test. A useful adjunct in the differential diagnosis of thrombocytopenia. AB - To aid in the rapid differential diagnosis of thrombocytopenia, the authors developed a latex agglutination test for glycocalicin, a proteolytic fragment of platelet membrane glycoprotein Ib. Plasma glycocalicin determinations were performed for 34 patients with thrombocytopenia. Plasma samples from four patients with aplastic anemia and ten patients with myelodysplastic syndromes, all with glycocalicin levels less than 0.6 mg/L by an enzyme-linked immunosorbent assay, all had negative results by the latex test. In contrast, positive latex agglutination titers were obtained for all 12 patients with idiopathic thrombocytopenic purpura. Eight patients with liver cirrhosis and splenomegaly had elevated levels of plasma glycocalicin, and all of their plasma samples produced agglutination. This latex agglutination test for glycocalicin allows a rapid discrimination of thrombocytopenia caused by impaired platelet production from that caused by increased platelet destruction; thus, it is suitable for use as a screening test in a routine clinical laboratory. PMID- 8249900 TI - The frequency of hypomagnesemia in acute and chronic care hospitalized patients. PMID- 8249901 TI - Reversible dilated cardiomyopathy due to growth hormone deficiency. PMID- 8249902 TI - Point of care laboratory testing in the emergency department. PMID- 8249903 TI - Where did this come from? Identification of sample mix-ups by DNA testing. PMID- 8249904 TI - Laboratory medicine education in United States medical schools. AB - The Academy of Clinical Laboratory Physicians and Scientists conducted a survey of US medical schools to examine the current status of laboratory medicine education, estimate the amount of teaching time available in the curriculum, and identify the most effective teaching practices. Questionnaires were sent to department heads and course directors at 126 US medical schools. Replies were received from 120 schools (95%), 83 of which offered a total of 132 courses in laboratory medicine. Only 68 schools (57%) had required courses. Most of the elective courses (35 of 50; 70%) were in general clinical pathology. Lectures remained the most common teaching format, with or without laboratory sessions and workshops. Computer-assisted instruction was used in only 10 schools. Laboratory medicine courses were offered in all 4 years of medical school, with the majority (70 of 132; 53%) in the second year, often integrated with general pathology. Opinion was divided over the relative importance of laboratory medicine instruction in the preclinical versus clinical years. PMID- 8249905 TI - The evaluation of a portable clinical analyzer in the emergency department. AB - A recently available portable clinical analyzer (PCA), which examines sodium, potassium, chloride, glucose, urea nitrogen, and hematocrit levels on 60 microL of blood and calculates hemoglobin and osmolality levels within 2 minutes, was evaluated. Blood from 574 patients was drawn by emergency department staff, who immediately tested the samples with the PCA and transported them for plasma analysis on a reference analyzer in the clinical laboratory. Correlations between the PCA and the reference analyzer were as follows: R2 = 0.987 for urea nitrogen; R2 = 0.97, glucose; R2 = 0.937, K;R2 = 0.79, hematocrit; R2 = 0.751, sodium; and R2 = 0.689 for chloride. With its rapid turnaround, small sample requirement, and ease of operation, the PCA is most useful in an emergency department setting, where immediate access to clinically relevant laboratory testing is required in support of urgent clinical decision-making. PMID- 8249906 TI - Does the emergency department need a dedicated stat laboratory? Continuous quality improvement as a management tool for the clinical laboratory. AB - Using the principles of continuous quality improvement (CQI), the authors conducted a study in response to a request from the Department of Emergency Medicine, Los Angeles County, and University of Southern California Medical Center, Los Angeles, for a dedicated stat laboratory in the emergency department. The stat orders to test serum electrolyte, glucose, blood urea nitrogen, creatinine, amylase, and lipase levels, prothrombin time, and complete blood count (CBC) were evaluated. The study was done in two phases. First, a baseline on stat laboratory test turnaround time was established, and problems were identified. Then, the authors reexamined the turnaround after problems were addressed and a new laboratory information system was installed. In the first phase, median within-laboratory turnaround for chemistry tests was 61 minutes from the time the specimens arrived in the laboratory and 70 minutes for CBCs from the time of accessioning. Delay in physician review of the results (45 minutes) was the longest component of overall turnaround. The second phase of the study found that the median within-laboratory turnaround had improved to 36 minutes for chemical and 55 minute for hematologic tests. However, other preanalytic factors outside the control of the laboratory, such as collecting blood and sending the specimens to the laboratory, and postanalytic delays in physician acknowledgment of the results remained the major components of the perceived turnaround delays. In conclusion, stat laboratory service for the emergency department improved with CQI. The study suggested that resources required to establish a dedicated stat laboratory in the emergency department would be more beneficial if directed toward reducing the preanalytic delays. Further, CQI has great potential as a management tool for the clinical laboratory. PMID- 8249907 TI - A comparison of the Coulter STKS, Coulter S+IV, and manual analysis of white blood cell differential counts in a human immunodeficiency virus-infected population. AB - The white blood cell differential counts obtained by manual analysis and those obtained by automated analysis using Coulter STKS and Coulter S+IV cell counters (Coulter Electronics, Hialeah, FL) in a human immunodeficiency virus (HIV) infected population were compared. Linear correlation analyses showed the STKS to be less accurate than the S+IV in determinations of the lymphocyte and granulocyte count. This finding was confirmed by difference analyses and by determining the number of measurements that fell outside of 95% confidence limits. Similar analyses were also done on patients not known to be HIV infected, and there was no disparity found between the STKS and S+IV in that group. In the HIV-infected population, the discrepancy between the manual and automated granulocyte counts performed on the STKS, but not on the S+IV, increased with increasing red blood cell mean corpuscular volume. Physicians caring for, and investigators performing studies on, HIV-infected patients should be aware of the potential for falsely low granulocyte numbers when using the differential from the STKS. PMID- 8249908 TI - Evaluation of the Sysmex NE-8000. Automated hematology analyzer in a high-volume outpatient laboratory. AB - An evaluation of SYSMEX NE-8000 (Toa Medical Electronics Co., Ltd., Kobe, Japan) hematology analyzer, including its automated five-part white blood cell (WBC) differential count, was performed in a high-volume outpatient laboratory. Precision, mixing studies, stability, carry-over, and linearity were all within the limits stated by the manufacturer. Evaluation of the five-part WBC differential revealed excellent correlation with manual differential neutrophil and lymphocyte counts. Monocyte, eosinophil, and basophil data were acceptable given the known poor precision of 200 cell manual WBC differential counts when low percentages of individual cell types are present. Overall, flagging for morphologic abnormalities displayed a sensitivity of 87% and a specificity of 97%. The combination of the flagging system with the visual record provided by the NE-8000 WBC histogram represents an effective tool for separating normal from abnormal specimens, and allows considerable selectivity in the performance of follow-up manual WBC differential counts. PMID- 8249909 TI - A parallel evaluation of four automated hematology analyzers. AB - A parallel evaluation was performed on four automated hematology analyzers: the Celldyn 3000 (Unipath Corp., Mountain View, CA), the Coulter STKS (Coulter Electronics Inc., Hialeah, FL), the Sysmex NE-8000 (Baxter Healthcare Corp., McGaw Park, IL), and the Technicon H*2 (Miles Corp., Tarrytown, NY). The protocol included evaluation of the complete blood count and differential leukocyte count (DLC) parameters. The DLC evaluation was performed using the National Committee for Clinical Laboratory Studies H20-A protocol. Based on this evaluation, the authors could not identify a single instrument that was clearly superior to the others. Overall, the four instruments were found to be safe and effective for diagnostic use; however, there were areas in which their performance was less than optimal. Particular questions were raised regarding the clinical usefulness of instrumental "flags" to identify qualitative leukocyte abnormalities. The results are discussed in relation to the selection of instruments for specific clinical applications. PMID- 8249910 TI - Comparative study of spleen pathology in drug abusers with thrombocytopenia related to human immunodeficiency virus infection and in patients with idiopathic thrombocytopenic purpura. A morphometric, immunohistochemical, and ultrastructural study. AB - A morphometric and immunohistochemical study was performed to assess the spleen's alterations in patients with autoimmune thrombocytopenia and in drug abusers with thrombocytopenia (DAT) related to human immunodeficiency virus (HIV) infection. A total of 34 patients were included in the study: 20 DAT patients and 14 with idiopathic thrombocytopenic purpura (ITP). Twenty HIV-negative splenectomy patients without thrombocytopenia were included as controls. Spleen weight in DAT patients (323.25 +/- 149.96 g, mean + standard deviation) was significantly increased compared with the ITP (164.28 +/- 29.79 g, P < 0.0001) and control (175.50 +/- 49.14 g, P < 0.0001) groups. The mean diameter of lymphoid follicles in the spleens of DAT patients (446.83 +/- 99.16 microns, was significantly higher than in those of the control patients (370.87 +/- 55.30 microns, P = 0.019). In control patients' spleens, the number of platelets in Billroth's cords was significantly higher (59.54 +/- 32.72/10(4) microns 2) than in those of the DAT (2.13 +/- 1.42/10(4) microns 2, P < 0.0001) and ITP (P < 0.0001) patients. The number of macrophages and ceroid histiocytes per 10(4) microns 2 of red pulp was significantly increased in both DAT (5.14 +/- 1.90) and ITP (7.48 +/- 4.38) patients compared with the control patients (3.66 +/- 1.10, P < 0.0001) and P = 0.06, respectively), and in ITP patients compared with DAT patients (P = 0.0136). The number of granulopoietic precursors per 10(4) microns 2 of red pulp was higher in the spleens of DAT (1.41 +2- 1.46, P < 0.0001) and ITP (0.92 +/- 0.75, P < 0.0001) patients compared with those of the control group. Transmission electron microscopy studies demonstrated platelet phagocytosis by macrophages of Billroth's cords and presence of myeloid metaplasia in spleens of DAT and ITP patients. Immunohistochemical studies showed a depletion of CD4+ lymphocytes in the T zone of splenic white pulp and an increased number of CD8+ lymphocytes in red pulp of DAT patients' spleens compared with those of ITP and control patients. There were no significant alterations in dendritic reticular cell network in the DAT group compared with the ITP and control groups. PMID- 8249911 TI - Detection and species identification of mycobacteria in paraffin sections of lung biopsy specimens by the polymerase chain reaction. AB - The authors analyzed 25 paraffin-embedded lung biopsy specimens for mycobacterial DNA by the polymerase chain reaction (PCR) from patients with pulmonary mycobacterial infection demonstrated by acid-fast stain, culture, or both. DNA was extracted from 4 microM unstained paraffin sections by proteinase K digestion followed by freeze-fracturing and amplified by nested PCR with primers for the mycobacterial 65-kDa antigen gene. Mycobacterial DNA was detected in 7 of 7 wedge and 9 of 18 transbronchial biopsy specimens by PCR. Nested PCR with direct visualization on an agarose gel was as sensitive as Southern blot hybridization. Serial dilution studies demonstrated that nested PCR could detect DNA amplified from 4-8 acid-fast organisms from a paraffin section. Restriction enzyme digestion of the amplified PCR product differentiated Mycobacterium tuberculosis from Mycobacterium avium-intracellulare. Polymerase chain reaction can detect low numbers of acid-fast organisms in paraffin sections and confirm and presumptively speciate mycobacterial infection when cultures are negative or not obtained. PMID- 8249912 TI - Relationship between intrahepatic expression of hepatitis B viral antigens and histology in Chinese patients with chronic hepatitis B virus infection. AB - Hepatic cellular and subcellular expression of hepatitis B virus (HBV) antigens- HBsAg, HBcAg, and HBeAg--in 143 Chinese patients with chronic HBV infection were studied by immunohistochemical techniques. Nuclear expression of HBcAg and nuclear and cytoplasmic expression of HBeAg showed a secular trend decreasing from the carrier state and chronic persistent hepatitis (CPH), through chronic active hepatitis (CAH), to cirrhosis with or without hepatocellular carcinoma. In contrast, cytoplasmic HBcAg expression was significantly greater in patients with CPH and CAH (P < .0108). In addition, cytoplasmic HBcAg correlated significantly with lobular activity, portal inflammation, and hepatitic activity (P < or = .007). Expression of cytoplasmic HBcAg also exceeded cytoplasmic HBeAg and nuclear HBcAg in liver specimens showing significant damage (P < .038). HBsAg, however, showed no secular trend and was not related to liver histology. The authors' findings support the theory that HBcAg is also the viral target antigen for immune-mediated liver damage in Chinese patients with chronic HBV infection. PMID- 8249913 TI - One century of mammary carcinoma in situ. What have we learned? PMID- 8249914 TI - Detection of Epstein-Barr viral genome in tumor cells of Warthin's tumor of parotid gland. AB - The presence of Epstein-Barr virus (EBV) DNA-sequences in specimens of Warthin's tumor in the parotid gland was studied. Using an in situ hybridization technique, EBV genome was detected in distinct cell types in routinely processed tissues. The authors' results indicated that EBV is frequently present in the cytoplasm (possibly because of a chronic, productive infection) of neoplastic cells of multiple/bilateral Warthin's tumor (MBWT) (86.7%), and only occasionally of solitary Warthin's tumor (SWT) (16.7%). In addition, EBV genome has often been documented in the cytoplasm of ductal cells (75% of MBWT and 33.3% of SWT), and occasionally of acinar cells (16.7% of both MBWT and SWT) of residual normal salivary gland tissue surrounding both MBWT and SWT. The almost constant presence of EBV DNA in tumor cells of MBWT, as opposed to the occasional finding in SWT, suggested a strong association between infection of cells with this virus and the development of multiple/bilateral lesions. The positivity of ductal epithelium supported speculations about the role of ductal cells as a histogenetic precursor of the tumor. This was the first report on the detection of EBV genome in Warthin's tumor. PMID- 8249915 TI - Identification of mismatched fixed specimens with a commercially available kit based on the polymerase chain reaction. AB - Specimen mix-ups inevitably occur and have the potential for great harm. The ability to investigate mix-ups objectively and assign fixed tissues to patients correctly is unfortunately limited, as most such assays require fresh specimens. A commercial kit based on the polymerase chain reaction (PCR) can be applied to the molecular genetic analysis of fixed tissues. This kit, which can amplify and distinguish 21 different genotypes at a polymorphic human leukocyte antigen (HLA) locus, was applied to investigate 16 cases of potential specimen mismatches. The majority of tissues were small and essentially irreplaceable biopsy specimens, and four cases involved minute fragments of potential "floaters." Data were successfully obtained from all 16 cases despite the collection from several different hospitals and the small quantities of tissue. The assay required approximately 2 days for completion; therefore, data were returned within a clinically useful time period. This study provided evidence that molecular genetic assays based on the PCR can be applied to routinely obtained fixed-tissue specimens to investigate potential mismatches. PMID- 8249916 TI - Cardiac myxoma. A clinicopathologic study. AB - The clinicopathologic features of 107 cardiac myxomas are presented. Eighty-one were in the left atrium, 18 in the right atrium, and 8 were biatrial or located in the ventricles. The mean age at presentation was 50 years and did not vary significantly by sex, race, or site in the left or right atrium. Histologically, 41% of tumors had surface thrombus; 41%, fibrosis; 23%, mitotic activity; 20%, calcification; 17%, gamma bodies; 8%, ossification; 7%, extramedullary hematopoiesis; 3%, mucin-forming glands; 3%, atypical cells simulating malignancy; and 1% had thymic rests. A majority of myxomas was immunohistochemically positive for QBEnd throughout the lesion, but in most tumors, factor-VIII related antigen was limited to the surface cells. A minority of myxomas stained for antismooth muscle actin and Ulex europaeus; KP-1 and cytokeratin were always negative. Right atrial tumors were more likely calcified than those in the left atrium. Embolic tumors were less often fibrotic than nonembolic myxomas and were more likely thrombosed and extensively myxoid with an irregular frond-like surface. Fibrotic and non-thrombosed tumors had a longer mean duration of clinical symptoms and were found in older persons. Recurrent, multiple, and familial myxomas were more often found in younger women and, more likely irregular surfaced and histologically myxoid; however, they were not likely to be histologically atypical, cellular, or incompletely excised. Five tumors caused the death of the patient by emboli to the brain or heart, but none metastasized. The histology of myxomas varied by site, clinical presentation, patient age, and immunohistochemical findings. PMID- 8249917 TI - A quantitative immunohistochemical evaluation of lentigo maligna and pigmented solar keratosis. AB - Pigmented solar keratosis (PSK) is sometimes clinically indistinguishable from lentigo maligna, a form of malignant melanoma in situ. Occasionally histologic diagnosis is also difficult. Accurate diagnosis is essential, as the treatment and prognosis for each condition differs considerably. To determine whether there was a significant overlap in the number of melanocytes in these sun-damaged skin lesions, or whether immunohistochemistry might be helpful in the differential diagnosis, the authors examined skin biopsy specimens from 26 patients with obvious lentigo maligna and 15 patients with PSK using 3 monoclonal antibodies (HMB-45, NK1C3, and vimentin) and 1 polyclonal antibody (S-100 protein). Formalin fixed paraffin sections were immunostained with each of the above antibodies, and immunopositive cells per mm2 of epidermis were counted. The difference between lentigo maligna and PSK counts was statistically significant at a level of P < .0001; furthermore, there was almost no overlap between the two groups. The sensitivity for the diagnosis of lentigo maligna was high with all antibodies. However, HMB-45 had the highest sensitivity and the lowest false-positive rate and was visually most pleasing. Using a cut-off count of 60 cells per mm2 of epidermis, HMB-45 had a sensitivity of 96% and a 0% false-positive rate. In this study, lentigo maligna was easily differentiated from PSK. The real value of immunohistochemistry in the differential diagnosis of these pigmented lesions should be tested in a prospective study using cases that are difficult to diagnose by routine light microscopy. PMID- 8249918 TI - Mutation analysis of K-ras oncogenes in gastroenterologic cancers by the amplified created restriction sites method. AB - A rapid, simple, and nonradioactive method for diagnosing point mutations of c-K ras oncogenes in gastroenterologic cancers is described. This method involved the selective amplification of DNA fragments from cancer tissues of surgical specimens with specific oligonucleotide primers, followed by digestion with restriction enzymes that recognized artificially created or naturally occurring restriction sites. To detect codon 12 mutations, an artificial Msp I site was created by introducing a single nucleotide mismatch into the 5' mutagenesis primer. Using a similar approach, an Hae III site was created to detect codon 13 mutations. Bal I and MBo II sites were used to detect codon 61 mutations. A total of 61 gastroenterologic cancer cases were studied. Of 35 cases of colorectal cancer, 7 showed mutations: 6 at codon 12 and 1 at codon 13. In 1 of 2 cases of cholangiocellular carcinoma, point mutation at codon 12 was found. One case of duodenal cancer showed point mutation at codon 12. No mutations were found in the cases of hepatocellular carcinoma (4), gastric cancer (12), esophageal cancer (3), or pancreatic cancer (2). PMID- 8249919 TI - Osteitis fibrosa cystica simulating metastatic tumor. An almost-forgotten relationship. AB - The diagnosis of primary hyperparathyroidism most often results from the incidental finding of hypercalcemia. In two recent cases of osteitis fibrosa cystica (OFC), however, patients without adequate access to health care served as graphic reminders that the clinical spectrum of the disease includes bone disease, and that OFC can be the presenting manifestation of long-standing primary hyperparathyroidism. Both patients complained of bone pain and had widespread osteolytic bone lesions in addition to hypercalcemia on a multitest biochemical panel. The presumptive diagnosis of malignancy with bone involvement (metastatic cancer or multiple myeloma) led to random bone marrow trephine biopsies. Examination of the bone marrow biopsy material revealed the characteristic pathology of OFC, leading to appropriate diagnosis and surgical management of large parathyroid adenomas in both patients. PMID- 8249920 TI - Intrauterine infarction of a pulmonary segment in a full-term newborn. A case report. AB - Intrauterine pulmonary infarction is extremely rare. In a series of 775 stillborn and newborn infants, only a few cases of acute pulmonary infarction were reported. The authors present the findings from a full-term infant who died at 9 hours of age of a chronic pulmonary infarction. Several pathogenic mechanisms, including the entry of placental thromboplastin into the fetal circulation, are discussed. PMID- 8249921 TI - Liver dysplasia and the interpretation of image and flow cytometric data of hepatocytes. PMID- 8249922 TI - Report of a case of localized Castleman's disease with progression to malignant lymphoma. PMID- 8249923 TI - A rare lymphoepithelial cyst of the pancreas. PMID- 8249924 TI - Comments on incisor torque control. PMID- 8249925 TI - Comment on static magnetic field bioeffects. PMID- 8249926 TI - Comment on computer article. PMID- 8249927 TI - Comment on "Straight wire: the next generation". PMID- 8249929 TI - Longitudinal shape changes of the nasal dorsum. AB - This investigation quantifies childhood and adolescent growth changes of the upper and lower nasal dorsum and evaluates various aspects of the persons' morphology that relate to shape changes of the dorsum. A longitudinal sample of 37 French-Canadian girls, each having cephalograms at 6, 10, and 14 years of age, was evaluated. The 6- to 10-year interval was chosen to represent childhood growth; the 10- to 14-year interval represented adolescent growth. The upper dorsum rotates upward and forward (counterclockwise) approximately 10 degrees between 6 to 14 years of age. The lower dorsum shows both downward and backward (clockwise) and upward and forward (counterclockwise) rotation average childhood and adolescent changes in angulation were not significant. The results clearly indicate that changes in the nasal dorsum are most closely related to angulation changes of the lower dorsum, particularly during adolescence. The lower dorsum rotates downward and backward in persons who show greater vertical and less horizontal growth changes. Rotational changes of the lower dorsum are most closely related with vertical changes at pronasale. PMID- 8249928 TI - Reflections on my involvement in orthodontic research. AB - Some years ago, Professor Takihashi, who was then the greatest living professor of orthodontics in Japan, called on us in Ann Arbor just before his retirement. I asked Dr. Takihashi who would replace him? and he replied, "There's a young man who will do better than I because he will be able to take Japanese orthodontics to the world and bring the world's orthodontics to Japan." That young man was Fujio Miura. Fujio Miura has, in the years since Takihashi spoke to me, written one of the most amazing records in the history of our specialty. His research is as far ranging as anyone's. For example, in biology he has written well on the ethnology of Japanese craniofacial morphology and that of the aborigines in Japan. He has written on developmental biology and on the tissue reaction of tooth movements. In metallurgy he has contrived alloys that have revolutionized our appliances, for he was a pioneer in the use of nickel-titanium wires. Fujio Miura invented direct bonding. Professor Miura has fulfilled Professor Takihashi's predictions well for he has become the most famous orthodontic professor in Japan's history. It is interesting to see that his students occupy most of the chairs of orthodontics in Japan and many elsewhere. Miura is an accomplished artist who has exhibited his work. This personable scholar probably attends more meetings of the American Association of Orthodontists Society than most American members, and he is equally at home in the European society, and many other orthodontic groups in the world. PMID- 8249930 TI - Long-term stability and prediction of soft tissue changes after LeFort I surgery. AB - Many evaluations of soft tissue changes after orthognathic surgery have been undertaken, and many correlations of soft tissue to hard tissue movements have been established. These studies have not, however, specifically discussed the long-term stability or characteristics of the soft tissue changes. The objectives of this study were (1) to determine the long term stability of soft tissue changes 5 years after LeFort I osteotomy, (2) to determine reliable correlations, if any, of soft tissue changes to bony movements effected in surgery, and (3) to determine the predictability of soft tissue changes as an aid to orthodontic treatment planning. Cephalometric data from 25 patients who were treated with LeFort I osteotomy with or without a concurrent mandibular procedure were analyzed retrospectively. Cases were selected from the patient records of the Department of Oral Surgery of the Vrije Universiteit in Amsterdam, the Netherlands. These patients were followed up at four time points, the last being a mean of 6.1 years after surgery. Analysis of stability data revealed that most horizontal and vertical soft tissue change after LeFort I surgery occurred in the first year after surgery. Significant (> 10%) change continued to occur for subnasale, labrale inferius, upper lip protrusion, lower lip protrusion, and soft tissue convexity during the subsequent 5 years. Hard tissue to soft tissue correlations were calculated and ratios of soft tissue to hard tissue movement were determined for appropriate hard and soft tissue landmarks at four time intervals. Reliable correlations of hard tissue change at surgery to 5-year soft tissue change could be made for 10 variables, which was considerably less frequently than for 1-year soft tissue change. The relatively low reliability of long-term prediction correlations suggests that soft tissue movements may be more independent of hard tissue over time. One-year prediction values were similar to 5-year values and thus could be used for prediction purposes in orthodontic treatment planning. Most short-term hard to soft tissue correlations found in the present study were in the range of those established by previous authors. Long term hard tissue to soft tissue correlations gave higher ratios of soft tissue movement secondary to maxillary surgery, approaching ratios of 1.0:1.0 for some variables. This finding, as well as the long-term stability data, suggest that soft tissue settling or equilibrium after surgery may take several years to complete. PMID- 8249931 TI - Molar and incisor changes with Wilson rapid molar distalization. AB - The purpose of this study was to determine the magnitude and direction of maxillary and mandibular first molar and incisor changes that occur during Class II molar correction with Wilson "rapid molar distalization." Nineteen patients (13 females, 6 males) were selected from the clinic at Loma Linda University Department of Orthodontics. All patients received maxillary bimetric distalizing arches (BDA), and either a mandibular three-dimensional lingual arch, with or without a passive 0.016 x 0.016-inch utility arch, or a traditional edgewise full banded and bonded arch. The arches were activated with open coil spring and Class II intermaxillary elastics. A total of five activations were completed or until visual inspection revealed a Class I molar relation. Prestudy and poststudy lateral cephalometric radiographs were taken with molar bands cemented. Dental changes were determined, with Ricketts superimpositions. The mean change in molar relation measured at the occlusal plane was 4.1 mm. The mean maxillary molar distalization was 2.16 mm, with 7.8 degrees of tip. The rate of maxillary molar movement was 0.56 mm per month. The mean mandibular molar mesial movement was 1.38 mm. The maxillary incisors protruded a mean of 0.3 mm, and extruded a mean of 1.6 mm. Except for the edgewise sample, all mandibular molars moved mesially further than the incisors, reducing the mandibular arch length. The presence of erupted maxillary second molars did not correlate with the rate of maxillary first molar movement, magnitude of movement, or amount of tipping that occurred. PMID- 8249932 TI - Lower third molar development subsequent to second molar extraction. AB - Development and eruption of lower third molars was examined in 63 subjects who were treated by extraction of second molars. Changes in mesiodistal and buccolingual lower third molar angulation were measured on 60 degrees cephalograms taken before extraction and 3 or more years later. The final position of lower third molars was assessed on models at the end of the observation period by using a scoring system. All lower third molars erupted in periods ranging from 3 to 10 years after extraction. Ninety-nine percent of the third molars upright mesiodistally, but few became as upright as the second molars they replaced. Model analysis showed that 96% of the lower third molars erupted in good or acceptable positions. PMID- 8249933 TI - A modified direct technique versus conventional direct placement of brackets: in vitro bond strength comparison. AB - Bond strength and failure location were evaluated in vitro for two methods of direct bracket bonding. Sixty human premolars were divided into two groups of 30 each. In group I the brackets were bonded with a two-paste adhesive by using the conventional direct method. In group II brackets were bonded with a newly developed modified direct technique. During the modified direct technique, unfilled resin catalyst liquid was applied to a bracket, which had a coating of hardened composite cured against a dental anatomic matrix (tooth). After the acid etched tooth was coated with unfilled resin base liquid, the bracket was placed. Mixed unfilled resin liquid (sealant) was then placed at the periphery of the bracket/tooth interface. Thus the major modification of the direct technique would entail fabrication by the manufacturer of a bracket with prehardened bis GMA composite resin on its backing. Bond strengths were 155.2 (SD = 35.7) and 140.6 (SD = 30.1) kg/cm2 for conventional and modified techniques, respectively. With the conventional method, failure occurred mainly at the tooth/composite interface. Failure seen with the modified technique was mixed, but the major mode was composite/bracket. Therefore this modified bonding method promises similar bond strengths and some advantages over the conventional method including, elimination of composite flash from around the brackets, ample working time, consistent adhesive thickness, and reduction of porosity. PMID- 8249934 TI - Natural cervical inclination and craniofacial structure. AB - The purpose of this study was to search for the statistical associations between postural and morphologic variables of the head. Interpretation of the facial structure was made by using both intracranial and the extracranial reference lines. The sample comprised natural head posture (NHP) cephalograms of 106 dental students, aged 19 to 29 years. Results showed that, when the facial structure was evaluated by using a NHP analysis based on extracranial reference lines, it was associated with the inclination of the cervical column to the true horizontal. In addition, in the natural position of the head, inclination of the NSL reference was found to be associated with the vertical localization of sella turcica (r = 724, p < or = 0.001), rather than the "extension" or "flexion" of the head. It was concluded that associations between posture and structure of the head are merely caused by the functional factors related to "forward cervical posture" and "vertical cervical posture". PMID- 8249935 TI - Facial plastic surgery, temporomandibular disorders, and orthodontics: a note. AB - The complexities introduced into the practice of orthodontics with the advent of the newer orthognathic surgery procedures are now being better understood. Widespread discovery of temporomandibular disorders further complicates this situation. Lack of understanding about dental problems and practice by some physicians may result in less than satisfactory patient care. Two case reports are presented that illustrate these problems. PMID- 8249936 TI - The orthodontic practice ownership transition. AB - Any orthodontist contemplating a transfer of ownership interests would be well served ensuring that the decision-making process is handled objectively and analytically and that appropriate advice is acquired before entering into an arrangement. Boiler plate agreements and rules of thumb do not work any more effectively than developing a single diagnosis and treatment plan for all patients. PMID- 8249937 TI - Unorthodox approaches to health care. PMID- 8249938 TI - Orthodontic bracket recycling. PMID- 8249939 TI - The adrenal incidentaloma. A pediatric perspective. AB - In our commentary, we have reviewed the literature on adrenal incidentalomas and specifically have tried to relate it to the pediatric population. The studies of others have shown that adrenal tumors are very common in patients with CAH (homozygote and heterozygote) and that these tumors are very rarely malignant and therefore should not routinely be surgically removed. In fact, we suggest that only under very rare circumstances is surgical removal justified. Perhaps, in the future, biochemical markers will allow us to distinguish between adrenal adenomas and carcinomas. We have tried to give broad guidelines for the care of patients with CAH with adrenal tumors, but these are guidelines only--they are not laws chiseled in stone. Finally, as radiological imaging is done more frequently and becomes even better at finding small adrenal tumors, this general topic will become increasingly relevant. We hope our thoughts will lead to lively discussion and useful studies in this "newly discovered" medical dilemma. PMID- 8249940 TI - A room of one's own and the Pogo factor. More women in pediatrics. PMID- 8249941 TI - Obturator internus muscle abscess: a case report and review of the literature. PMID- 8249942 TI - The female adolescent athlete and iron. PMID- 8249943 TI - Identification and treatment of nonsteroidal anti-inflammatory drug-induced gastroduodenal injury in children. PMID- 8249944 TI - Aspirin and bismuth subsalicylate. PMID- 8249945 TI - The genetics of autoimmune diabetes. Approaching a solution to the problem. AB - Molecular genetics has had a substantial impact on our understanding of inherited susceptibility to insulin-dependent diabetes mellitus. Alleles at the HLA-DQA1 and DQB1 loci appear to have the greatest influence on diabetogenesis. Other promising loci are present on chromosome 11 in the vicinity of the insulin gene. We have sought not only to improve our prediction of insulin-dependent diabetes mellitus but also to reveal the underlying immune and nonimmune defects that predispose to autoimmune beta-cell destruction. Continued advances in the field of genetics will aid in the prevention of insulin-dependent diabetes mellitus, which is our ultimate goal. PMID- 8249946 TI - Miller-Dieker syndrome. Detection of a cryptic chromosome translocation using in situ hybridization in a family with multiple affected offspring. AB - OBJECTIVE: To describe a family in whom fluorescence in situ hybridization allowed for accurate diagnosis of Miller-Dieker syndrome in an at-risk pregnancy and determination of parental carrier status. DESIGN: Retrospective case analysis and application of a new molecular tool to evaluate the family. SETTING: Health maintenance organization. The family was followed up by the Departments of Medical Genetics, Pediatrics, and Obstetrics and Gynecology, Kaiser Permanente Medical Center, Panorama City, Calif. PARTICIPANTS: Members of a single family. INTERVENTIONS: Clinical evaluation and neuroimaging studies of the proband. Prenatal diagnosis via ultrasonography and amniocentesis. Chromosomal evaluation of the couple and their offspring. In situ hybridization studies in both parents and an affected fetus. MEASUREMENTS/MAIN RESULTS: We describe a family in whom fluorescence in situ hybridization detected a submicroscopic deletion of the Miller-Dieker syndrome critical region 17p13.3 arising from a cryptic translocation in one of the parents. The proband was determined at birth owing to the presence of multiple congenital anomalies, including low birth weight, microcephaly, agenesis of the corpus callosum, lissencephaly, cerebral atrophy, unilateral ptosis, polydactyly, and omphalocele. High-resolution chromosome banding analysis findings were normal in the parents and proband, who died at age 4 years. There were four subsequent pregnancies: two ended in first-trimester spontaneous abortion, and in the other two, large omphaloceles were detected in fetuses at 15 and 13 weeks' gestation. Both pregnancies were terminated. Fluorescence in situ hybridization probes for 17p13.3 had become available before the most recent pregnancy and were used to study parental and fetal cells. As a result, a balanced cryptic translocation between chromosome 17 and chromosome 19 was identified in the father: 46,XY,t(17;19)(p13.3q13.33). An unbalanced form of the translocation, involving a deletion of 17p13.3, was detected with fluorescence in situ hybridization in the fetus. This finding was in accordance with a clinical diagnosis of Miller-Dieker syndrome. CONCLUSIONS: Molecular cytogenetic technology should be used in cases of suspected Miller-Dieker syndrome when high-resolution cytogenetic analysis fails to detect del(17) (p13.3). Positive findings should be followed up with parental studies. In addition, omphalocele should be included among the list of malformations that make up the Miller-Dieker syndrome. PMID- 8249947 TI - Risk of congenital heart defects in relatives of patients with atrioventricular canal. AB - OBJECTIVE: To investigate the overall occurrence of congenital heart disease in 103 pedigrees with a proband affected with atrioventricular canal. DESIGN: Family study of patient series. SETTING: Department of Pediatric Cardiology, Bambino Gesu' Hospital, Rome, Italy. PARTICIPANTS: One hundred three consecutive patients with atrioventricular canal and normal karyotype and their first- and second degree relatives. INTERVENTIONS: None. MEASUREMENTS/MAIN RESULTS: In 12 pedigrees (11.7%), one or more relatives had concordant or discordant congenital heart disease. Congenital heart disease occurred in four (1.9%) of the 206 parents of probands, in four (3.6%) of the 111 siblings, and in five (0.8%) of the 644 uncles or aunts. None of the grandparents had congenital heart disease. Atrioventricular canal affected several family members in five pedigrees. CONCLUSIONS: Familial aggregation of atrioventricular canal in several pedigrees requires careful collection of family histories, accurate cardiologic evaluation of the first- and second-degree relatives of the probands, and fetal echocardiography in all pregnant women in at-risk families. PMID- 8249948 TI - Evidence of a major gene with pleiotropic action for a cardiovascular disease risk syndrome in children younger than 14 years. AB - OBJECTIVE: To test the plausibility of the existence of a genetically based syndrome involving obesity, hypertension, and a central deposition of body fat. DESIGN: Survey of a random stratified sample of the US population. PARTICIPANTS: Male and female children aged 13 years or younger (mean, 5.3 years; SD, 3.7 years; median, 4.0 years; range, 6 months to 13 years) were chosen from National Health and Nutrition Examination Survey II data. INTERVENTIONS: None. MEASUREMENTS/MAIN RESULTS: Using multivariate commingling analysis, we evaluated the hypothesis that a major gene produces a syndrome involving the phenotypic indicators of body mass index (in kilograms per square meter), subscapular-to triceps skinfold thickness ratio, systolic blood pressure, and diastolic blood pressure. Maximum likelihood estimation was used to test competing models. A model with three component distributions and unequal variance-covariance matrices fit significantly better than any competing model. CONCLUSIONS: Our findings support the existence of a distinct cardiovascular disease risk syndrome in children and suggest that it may be the result of a major gene with pleiotropic effects. PMID- 8249949 TI - X-linked lymphoproliferative disease. Detection of a paternally inherited mutation in a German family using haplotype analysis. AB - OBJECTIVE: To study the inheritance of the X-linked lymphoproliferative disease (XLP) locus in a German family. DESIGN: Haplotype segregation analysis. SETTING: Departments of Pediatrics and Human Genetics, University of Wurzburg and University of Ulm, Federal Republic of Germany. PARTICIPANTS: Fourteen members of a family with XLP. INTERVENTIONS: None. MEASUREMENTS/MAIN RESULTS: Using molecular genetic techniques, we diagnosed the XLP status of the members of a German family. Restriction fragment length polymorphism analysis with closely linked polymorphic X-chromosomal DNA markers (Xq25-q27) revealed XLP carrier status in a female infant. Moreover, the XLP mutation was suggested to have occurred in the germ cell of the grandfather. CONCLUSION: This is the first report of a paternally inherited XLP mutation. The recurrence risk for germ cell mosaicism in XLP may be similar to that of X-linked Duchenne muscular dystrophy. PMID- 8249950 TI - Cytogenetic survey of Apert syndrome. Reevaluation of a translocation (2;9)(p11.2;q34.2) in a patient suggests the breakpoints are not related to the disorder. AB - The association of Apert syndrome with a translocation (2p-;Cq+) was previously reported in this journal. On reexamination using high-resolution chromosome banding, results showed both the patient and her unaffected father carry the balanced translocation (2;9)(p11.2;q34.2). This finding suggests the rearrangement is unlikely to be the cause of her disorder. Other chromosomal anomalies and genes known to be located at or near these breakpoints and a cytogenetic survey of patients with Apert syndrome are reviewed. PMID- 8249951 TI - Autosomal recessive microcephaly, microcornea, congenital cataract, mental retardation, optic atrophy, and hypogenitalism. Micro syndrome. AB - Three affected children from an inbred family had microcornea, microcephaly, congenital cataract, severe mental retardation, retinal dystrophy, optic nerve atrophy, hypothalamic hypogenitalism, and agenesis of the corpus callosum. The disorder is presumably autosomal recessive; no identical syndrome has been described, but we consider syndromes with similar features. PMID- 8249952 TI - Effects of methylphenidate on sleep in children with attention-deficient hyperactivity disorder. An activity monitor study. AB - OBJECTIVE: To assess the effects of methylphenidate hydrochloride on sleep patterns in children diagnosed as having attention-deficit hyperactivity disorder (ADHD). DESIGN: A double-blind, controlled drug-placebo cross-over design. SETTING: Home. SUBJECTS: Ten children (aged 6 years 9 months to 12 years & months) diagnosed as having ADHD were consecutively recruited and compared with age- and sex-matched normal controls. TREATMENT: Methylphenidate hydrochloride (0.3 to 0.4 mg/kg) or placebo was administered at 7:30 AM. MEASUREMENTS AND RESULTS: Each child underwent activity monitoring at home during 6 days of no treatment (baseline) followed by placebo and methylphenidate treatment. The results of the three trial stages, as well as those of the 20 age- and sex matched normal controls, were compared. A shorter total sleep duration was evident during the methylphenidate treatment compared with that of baseline and placebo treatment. The amount of quiet sleep was lower (however, not significantly) among the study group compared with controls, whereas no such difference was noted during methylphenidate treatment. Night-to-night sleep pattern stability was found. No other differences were found either between children with ADHD and controls or between on and off stages of methylphenidate treatment. CONCLUSIONS: These results support the notion that ADHD is a centrally generated disorder attributable to hypoarousal, which subsequently stimulates motor overactivity. Methylphenidate does not appear to affect sleep patterns adversely and possibly normalizes them in patients with ADHD. PMID- 8249953 TI - Passive-active immunization in infants of hepatitis Be antigen-positive mothers. Comparison of the efficacy of early and delayed active immunization. AB - OBJECTIVE: To assess the efficacy of late active immunization against hepatitis B concomitant with diphtheria, pertussis, tetanus, and polio vaccine in high-risk infants receiving hepatitis B immune globulin at birth. DESIGN: Randomized study of infants born to mothers positive for hepatitis B surface antigen (HBsAg) and hepatitis Be antigen (HBeAg). SETTING: Three large city hospitals and one rural area providing prenatal care and obstetric services. SUBJECTS: Eighty neonates of HBsAg- and HBeAg-positive carrier mothers received 0.5 mL/kg of body weight hepatitis B immune globulin within 2 hours of birth and hepatitis B vaccine (10 micrograms) at 0, 1, 2, and 11 months of age (group A) or at 3, 4, 5, and 11 months of age concomitant with diphtheria, pertussis, tetanus, and polio immunization (group B). A second dose of hepatitis B immune globulin was given to infants on schedule B at 3 months. MAIN OUTCOME MEASURES: Blood samples were collected at 0, 3, 6, 11, and 12 months of age and tested for antibodies against hepatitis B core antigen and HBsAg. Follow-up visits were scheduled annually up to 5 years of age. RESULTS: Eight infants were excluded from analysis. During the study period, six children became HBsAg carriers, three in each group, which corresponds to a 5-year incidence of infection of 9% and 8% for groups A (three of 35) and B (three of 37), respectively. Subclinical infections (persistent anti HBc positivity beyond month 12 or appearance of anti-HBc) were encountered in another eight infants (four in each group). CONCLUSION: Late active immunization starting at 3 months of age appears to provide similar protective efficacy as active immunization starting at birth when combined with hepatitis B immune globulin at 0 and 3 months of age. PMID- 8249954 TI - Adrenal tumor complicating untreated 21-hydroxylase deficiency in a 5 1/2-year old boy. AB - OBJECTIVE: A 5 1/2-year-old boy presenting with virilization was diagnosed as having classic 21-hydroxylase deficiency complicated by an adrenal tumor. We attempted to document a reduction in the size of the tumor with glucocorticoid therapy. DESIGN: Case study. SETTING: Referral center. INTERVENTION: Glucocorticoid therapy was instituted for congenital adrenal hyperplasia. Surgery for the adrenal mass was deferred, and the size of the mass was monitored by serial ultrasonography. RESULTS: Baseline values of serum 17 alpha hydroxyprogesterone (186.6 nmol/L) and testosterone (24.7 nmol/L [7.1 ng/mL]) were elevated. After instituting steroid treatment, 17 alpha-hydroxyprogesterone was suppressed (13.0 nmol/L), and testosterone remained undetectable on follow up. However, the size of the mass increased during 6 months. Unilateral adrenalectomy performed at that time revealed a well-encapsulated adenoma in a hyperplastic gland. CONCLUSIONS: Untreated classic congenital adrenal hyperplasia may be complicated by an adrenal tumor even at a young age. Suppression of adrenal androgens by glucocorticoid therapy was not accompanied by regression of the tumor in our patient. PMID- 8249955 TI - Maltreatment of children born to cocaine-dependent mothers. AB - OBJECTIVE: To investigate the relationship between maternal cocaine dependency and child maltreatment in a cohort of young children. DESIGN: Historical cohort study at an urban, tertiary care medical center. PATIENTS: 47 infants, born between January and September 1989, whose mothers were regular users of cocaine during pregnancy, based on history and the results of newborn's urine toxicology screens. These cocaine-exposed infants were matched to a comparison group of 47 infants whose mothers did not use cocaine during pregnancy. Matching was on the basis of birth date, race, method of payment for the hospitalization, and marital status of the mother. MAIN OUTCOME MEASURES: Occurrence of maltreatment (physical abuse, sexual abuse, or neglect), and placement either in foster care or with a substitute caretaker. RESULTS: By 24 months of life, maltreatment had occurred in 23% of the cocaine group vs 4% of the comparison group (risk ratio, 5.5; 95% confidence interval, 1.3 to 23.5). Physical abuse had occurred in 11% of the cocaine group vs 2% of the comparison group, while neglect had occurred in 11% vs 0% (P < .05). Changes in placement had occurred in 20% of the cocaine group vs 2% of the comparison group (risk ratio, 10.0; 95% confidence interval, 1.3 to 75.1). Of the 10 placements, only three were directly linked to an episode of maltreatment. CONCLUSIONS: Children identified during the neonatal period as regularly "exposed" to cocaine in utero are at a substantially increased risk both of maltreatment and of changes in the primary caretaker during the first 24 months of life. PMID- 8249956 TI - Applicability of the Greulich and Pyle skeletal age standards to black and white children of today. AB - OBJECTIVE: The Greulich and Pyle skeletal age atlas was derived from white children of upper socioeconomic level during the 1930s. To our knowledge, the Greulich and Pyle standards have not been reassessed for both black and white children; it was the purpose of this study to reassess the applicability of these standards to today's children. DESIGN: A "blinded" review of hand roentgenograms taken for the evaluation of trauma was performed. Age of the subjects was evenly distributed between 0 and 18 years. The roentgenograms were scored for bone age by five individuals from different disciplines and levels of training. The difference between the median bone age and the child's chronologic age was calculated for each roentgenogram and stratified into four age groups: early childhood (0 to 4 years), middle childhood (4 to 8 years), late childhood (8 to 13 years), and adolescence (13 to 18 years). SETTING: The roentgenograms were obtained from four hospital emergency rooms in the Lake Erie basin area, the same geographic area from which the Greulich and Pyle standards originated. PATIENTS: There were 841 children: 452 boys and 389 girls, 461 black and 380 white children. RESULTS: The bone ages and chronologic ages were similar for white girls of all ages. Black girls were skeletally advanced by 0.4 to 0.7 year (P < .001), except during middle childhood. White boys were skeletally delayed during middle childhood by 0.9 year (P < .001) and during late childhood by 0.4 year (P < .01), but they were advanced during the adolescent years by 0.5 year (P < .01). Black boys showed no difference except for the adolescent group, which was skeletally advanced by 0.4 year (P < .02). CONCLUSIONS: The Greulich and Pyle atlas is not applicable to all children today, especially black girls. We should be aware of this information when making clinical decisions requiring accurate bone ages. PMID- 8249957 TI - Radiological case of the month. Proteus syndrome: benign angiolipomatous tumor with intraspinal extension. PMID- 8249958 TI - Pathological case of the month. Wells' syndrome. PMID- 8249959 TI - Picture of the month. Cutaneous leishmaniasis. PMID- 8249960 TI - Breast cancer and colorectal neoplasia: double jeopardy or not? PMID- 8249961 TI - Because it's there. PMID- 8249962 TI - Gallstones, cholecystectomy, and colorectal cancer. AB - PURPOSE: To examine the controversial association of gallstones, cholecystectomy and colorectal cancer. Methodologic explanations for the association include ascertainment bias, unequal diagnostic testing, and necropsy selection bias. Necropsy screening, which eliminates unequal diagnostic testing and ascertainment bias and reduces necropsy selection bias, was used to study this controversy. METHODS: Adult necropsies at the University of Kansas Medical Center from 1950 to 1984 were reviewed. Patients with colorectal cancer, gallstones, or who had cholecystectomy during life were excluded. The remaining patients were those in whom neither colorectal cancer nor gallstones were suspected during life (reducing selection bias). The occurrence of gallstones and colorectal cancer among these individuals was then determined (reducing ascertainment bias and unequal diagnostic testing). RESULTS: Of 7485 persons receiving necropsy, 239 had colorectal cancer diagnosed during life and an additional 604 had gallstones or cholecystectomy, leaving 6642 patients available for study. Overall, no association between colorectal cancer and gallstones was found. In women, gallstones were associated with colorectal cancer; 6/447 (1.3%) with gallstones had colorectal cancer compared with 11/2259 (0.4%) without gallstones who had colorectal cancer, p = 0.048, odds ratio 2.78 (95% CI 0.84-8.25). A stronger association was found between right-sided colorectal cancer and gallstones (odds ratio 6.79, 95% CI 1.14-46.46). CONCLUSIONS: These data suggest an association between gallstones and colorectal cancer among women. Gallstones may indicate patients at higher risk for colorectal cancer. Studies associating cholecystectomy with colorectal cancer may be explained--not by ascertainment bias--but, rather, by susceptibility bias. The reason for the cholecystectomy (gallstones) may be the correct association and not the cholecystectomy itself. PMID- 8249963 TI - Endoscopic management of primary sclerosing cholangitis: review, and report of an open series. AB - Of the multiple treatment options available to physicians managing patients with primary sclerosing cholangitis (PSC), no single form of therapy has proven to be efficacious or to improve the clinical outcome of affected patients. Surgical and radiological methods require multiple procedures which are associated with increased morbidity, the most serious being sepsis. Endoscopic therapy is safer and easier to perform, and has been shown to provide symptomatic as well as objective improvement in affected patients. It, too, is associated with some complications, although less serious than other alternative treatments, but it is not effective in protracted disease. The potential of overlooking or missing cholangiocarcinoma is more apparent when nonsurgical techniques are employed. In this review we will discuss the various endoscopic treatment modalities utilized in PSC disease, and include the results that have been obtained with these techniques since they were incorporated into our armamentarium in 1983. PMID- 8249964 TI - Colonic adenomas in asymptomatic women with a history of breast cancer. AB - One hundred ninety-three asymptomatic women with a personal history of breast cancer underwent screening colonoscopy. One hundred sixty-eight women had breast cancer as their only potential risk factor for colonic neoplasia, and 25 had a family history of colorectal neoplasia in addition to their personal history of breast cancer. Among women with breast cancer, increasing age and body weight were each predictive of an increasing prevalence of colonic adenomas. The prevalence of colonic adenomas in women aged 50-75 yr whose only potential risk factor was breast cancer was 18%, and was identical to the prevalence of colonic adenomas in 186 asymptomatic average-risk control women aged 50-75 yr (odds ratio 1.0, 95% CI 0.54-1.87). We conclude that a personal history of breast cancer does not predict a higher prevalence of colonic adenomas. PMID- 8249965 TI - Failure of providers to vaccinate HIV-infected men against hepatitis B: a missed opportunity. AB - OBJECTIVE: To carry out an audit of hepatitis B immunization practices in an outpatient HIV clinic. METHODS: We reviewed the medical records of all new HIV infected patients seen between October 1, 1990 and December 31, 1991. RESULTS: The 125 patients were men with a mean age and CD4 count of 43 yr and 240 cells/mm3, respectively. Fourteen percent (14%) of men who showed a clear need for vaccine, having no HBV markers, were not vaccinated by the clinic staff. Further, 16% whose susceptibility to HBV infection was unclear, with anti-HBc as a sole HBV marker, were not evaluated with a booster dose of hepatitis B vaccine in an attempt to elicit an anamnestic response. CONCLUSIONS: In failing to vaccinate or evaluate the 30% of patients without HBsAg or anti-HBs, our providers are missing an important opportunity in preventive medicine. We urge others to examine their own hepatitis B screening and vaccination practices. PMID- 8249966 TI - Colorectal cancer screening: is one-year surveillance sigmoidoscopy necessary? AB - OBJECTIVES: To assess the diagnostic yield of the 1-yr follow-up flexible sigmoidoscopy examination for the detection of colonic neoplasia in a colorectal cancer screening program. METHODS: Data were prospectively collected for flexible sigmoidoscopies performed between July 1991 and July 1992 as part of the University of South Alabama Colon Cancer Registry. RESULTS: A total of 217 subjects with an age-related risk of colon carcinoma and no evidence of occult blood by stool examination were enrolled. One hundred and sixteen underwent initial sigmoidoscopic examination, and 101 had a 1-yr surveillance examination. Ten of the 116 patients (8.6%) undergoing initial surveillance sigmoidoscopy were found to have neoplastic lesions. In contrast, only 1/101 (1%) patients undergoing 1-yr follow-up surveillance sigmoidoscopy was found to have a neoplastic lesion. Neoplastic lesions in both groups were adenomas. No carcinomas were found. CONCLUSIONS: These data suggest that the 1-yr surveillance sigmoidoscopy adds little to the diagnostic yield of colon cancer screening. PMID- 8249967 TI - Healing of transsphincteric perianal fistulas in Crohn's disease using a new technique. AB - OBJECTIVES: Transsphincteric perianal fistulas in Crohn's disease are often refractory to medical therapy and difficult to treat surgically. Our objective was to determine whether a new method of anal fistulotomy was effective in healing these fistulas. METHODS: In this new method, the internal opening of the fistula tract was displaced distally to the region of the anal mucosa in 26 patients with Crohn's disease. RESULTS: Healing was achieved within 5 weeks of surgery in 45/49 (92%) of the fistulas and was not influenced by perioperative medication, site of preexisting Crohn's disease, or condition of the rectum. There has been no alteration of preoperative continence. CONCLUSIONS: We conclude that this method is highly effective in treating transsphincteric perianal fistulas in Crohn's disease. PMID- 8249968 TI - Is there a role for upper gastrointestinal endoscopy in the evaluation of patients with occult blood-positive stool and negative colonoscopy? AB - OBJECTIVE: To determine the role of upper gastrointestinal endoscopy in the clinical investigation of patients with occult blood-positive and negative colonoscopy. METHODS: This is a retrospective study of 211 consecutive patients who were investigated by upper gastrointestinal endoscopy for occult blood positive stool and negative colonoscopy. Patients were categorized into four groups: asymptomatic (n = 117), symptomatic (n = 37), severe anemia (hemoglobin < 10 g/dl) (n = 33), and incomplete (CBC not obtained or insufficient documentation regarding the presence or absence of symptoms) (n = 24). RESULTS: Eighty-eight of the 211 patients (42%) had abnormal upper gastrointestinal endoscopic findings: 43% of the "asymptomatic" group, 35% of the "symptomatic" group, 45% of the "severe anemia" group, and 42% of the "incomplete" group. The number of patients with abnormal upper gastrointestinal findings increased with age (p = 0.0002). Furthermore, there was a significant difference in upper gastrointestinal lesions between the patients who were 60 yr of age or older, and the patients less than 60 yr of age: 51% compared to 22% (p = 0.00003). All upper gastrointestinal endoscopic findings were benign, with 53 of the 88 patients (60%) having erosive gastritis. However, only 25 of those 88 patients (28%) had a lesion for which a specific treatment was available. CONCLUSIONS: Upper gastrointestinal endoscopy in patients with occult blood-positive stool and negative colonoscopy has a very low yield for significant lesions, not only from a therapeutic standpoint, but also for detection of malignant lesions. Prospective studies are needed to determine which specific groups of patients deserve further investigation. PMID- 8249969 TI - Acute gastrointestinal bleeding from portal hypertensive gastropathy: prevalence and clinical features. AB - The clinical and endoscopic features of patients diagnosed with acute bleeding due to portal hypertensive gastropathy (PHG) were evaluated. Acute bleeding from PHG was diagnosed in 12 patients (0.8%) of 1496 patient admissions prospectively evaluated by our Gastrointestinal Bleeding Team over a 3-yr period, and accounted for 8% of nonvariceal bleeding diagnosed in patients with liver disease. The median age of PHG patients (8M:4F) was 66 yr (range, 37-72). The most common underlying liver disease was alcoholic cirrhosis (five patients). The majority of patients presented with melena. There was no hemodynamic instability. Six patients had prior sclerotherapy. Esophageal varices, grades 1 (five patients) and 2 (three patients), were present. Severe PHG was encountered in seven patients. There was no correlation between the presence or absence of varices, the grade of esophageal varices, and prior sclerotherapy on the severity of PHG or continued bleeding. The mean hospital stay was 6 days. An average of 4 units of blood was transfused per patient (range, 2-8). Continued bleeding occurred in nine patients (75%), one of whom had mild PHG. Two patients with continued bleeding subsequently were diagnosed with portal hypertensive vasculopathy distal to the stomach. There was one episode of encephalopathy and no related mortality. Acute (overt) bleeding from PHG is uncommon, likely to recur, and can evolve into a pattern of chronic blood loss. PMID- 8249970 TI - Atypical rectosigmoid histology in children with newly diagnosed ulcerative colitis. AB - BACKGROUND: In the untreated patient with inflammatory colitis, rectal sparing or patchy rectal inflammation is generally considered a sign of Crohn's disease (CD), rather than ulcerative colitis (UC). METHODS: The initial endoscopic rectosigmoid mucosal biopsies obtained at disease onset from 12 untreated children with UC who ultimately required surgery were blindly reviewed (randomly mixed with another 62 specimens obtained from children with CD or treated UC). Biopsies were classified as typical UC if there was diffuse, active inflammation and severe crypt destruction or distortion. Those with patchy, active inflammation and only mild crypt changes were classified as CD. Because all 12 subjects had ultimately been proven to have UC by examination of a subtotal colectomy specimen, for the purposes of this report biopsies read as either normal or CD were both considered evidence of atypical UC with rectal sparing. RESULTS: Five of 12 subjects (seven biopsies) had atypical histology. Mild, patchy inflammation was seen in six rectal or sigmoid biopsies, whereas one rectal biopsy was normal. The remaining seven subjects (10 biopsies) had diffuse inflammation. Two of five subjects with atypical biopsies had an endoscopically normal rectosigmoid, one had patchy inflammation, and the remaining two had diffuse endoscopic changes. All seven subjects with typical UC histology had diffuse endoscopic changes. Subjects with atypical findings could not be differentiated by age, duration, or types of symptoms at presentation, years of disease at colectomy, or indications for colectomy. CONCLUSIONS: Patchy or absent inflammation of the rectum and sigmoid can be present in untreated children with UC at disease onset. Because such children may be mistakenly diagnosed as having CD, these data must be considered when treatments or clinical research protocols are designed to include children with colitis. PMID- 8249971 TI - Acid secretion and serum gastrin in normal subjects and patients with duodenal ulcer: the role of Helicobacter pylori. AB - OBJECTIVES: To compare gastric secretory function in patients with duodenal ulcer and in healthy volunteers with and without Helicobacter pylori infection. METHODS: Basal acid output, peak acid output, meal-stimulated acid output, fasting and meal-stimulated serum gastrin concentrations were measured in 136 healthy volunteers (63 H. pylori positive, 73 H. pylori negative) and 52 duodenal ulcer patients, all but one of whom were H. pylori positive. RESULTS: By multivariate linear regression analysis, H. pylori infection was a significant negative predictor of basal acid output and a positive predictor of fasting and meal-stimulated gastrin concentrations. When compared to truly normal (i.e., H. pylori-negative) control subjects, duodenal ulcer patients had elevated basal acid output, peak acid output, fasting and meal-stimulated gastrin concentrations. CONCLUSIONS: Our results show that in patients with duodenal ulcer disease, hypergastrinemia is largely related to gastric H. pylori infection, whereas acid hypersecretion is due to factors other than H. pylori. PMID- 8249972 TI - Sugar malabsorption in functional bowel disease: clinical implications. AB - OBJECTIVE: To investigate the relationship of sugar malabsorption to the development of clinical symptoms in functional bowel disease. METHODS: Twenty five consecutive outpatients [five men, 20 women; mean age 38.7 +/- 2.6 (SEM) yr] with functional bowel disease and symptoms suggestive of carbohydrate malabsorption were studied. Twelve healthy subjects [six men, six women; mean age 35.7 +/- 3.7 (SEM) yr] acted as the control group. Sugar malabsorption was assessed by breath-hydrogen test after an oral load of various solutions containing lactose (50 g), fructose (25 g), sorbitol (5 g), fructose plus sorbitol (25 + 5 g), and sucrose (50 g). The severity of symptoms developing after sugar challenge was studied. In addition, the effect on clinical symptoms of a diet free of the offending sugars, compared to a low-fat diet, was assessed. RESULTS: Frequency of sugar malabsorption was high in both patients and controls, with malabsorption of at least one sugar in more than 90% of the subjects. Median symptom scores after both lactose [median 6; interquartile (IQ) range 3-7] and fructose plus sorbitol (median 2; IQ range 0-4) malabsorption were significantly higher than after sucrose load (median 1; IQ range 0-1.5) in functional bowel disease patients (p = 0.001 and p = 0.007, respectively). However, there were no differences in healthy controls. In addition, symptoms score after both lactose and fructose plus sorbitol malabsorption was significantly higher in patients than in control subjects (p = 0.02 and p = 0.008, respectively). On the other hand, H2 production capacity, as measured following lactulose load, was significantly higher in patients than in controls. The clinical symptoms improved in 40% of the evaluated patients after restriction of the offending sugars. CONCLUSIONS: These results suggest that sugar malabsorption may be implicated in the development of abdominal distress in at least a subset of patients with functional bowel disease. PMID- 8249973 TI - Clinical significance of Mallory-Weiss tears. AB - OBJECTIVES: To assess course and outcome of patients with endoscopically diagnosed Mallory-Weiss tear bleeding. SUBJECTS: Thirty-four subjects seen during a 5-yr period formed the study group. RESULTS: Available follow-up after index bleed was 27.5 months +/- 2.7 SEM. There was no antecedent explanation for the tear, such as nausea, retching, abdominal pain, or vomiting in 12/28 (42.9%). Hematemesis on first emesis was noted in 13/26 (50%). Mean transfusion requirements were 2.6 +/- 1.0 (SEM) units of packed cells (range, 0-28), and 9/34 (26.5%) received four or more units. Two had therapeutic endoscopy and three required surgery to control bleeding. Thirty-day mortality noted four deaths, all multiorgan system failure related to the bleed. Patients who died had other endoscopic abnormalities, such as ulcers or varices, and all had an alcohol history. Two patients of 20 contacted had recurrent bleeding. One had another tear. The other had intermittent recurrent bleeding and refused care. CONCLUSIONS: Mallory-Weiss tear bleeding may be significant and recurrent. It may cause death or require transfusion, therapeutic endoscopy, and surgery. PMID- 8249974 TI - A multicenter case-controlled study of percutaneous endoscopic gastrostomy in HIV seropositive patients. AB - OBJECTIVES: Malnutrition is an important cause of morbidity and mortality in AIDS patients. Percutaneous endoscopic gastrostomy (PEG) is a safe and effective method of providing nutrition in HIV-seronegative patients who are unable to swallow food, but have an otherwise functional alimentary tract. This study analyzes the risks and benefits of PEG in HIV-seropositive patients. METHODS: The risks of PEG were analyzed in 14 consecutive HIV-seropositive patients admitted to two university hospitals, and were compared with the risks in a sex- and age matched control group of 21 patients. Thirteen HIV patients had AIDS. RESULTS: PEG indications included mechanical esophageal obstruction in six, wasting in six, and central nervous system disorders in two. The mean weight of the HIV patients increased by 7.4% +/- 3.0% (SE) 3-8 wk after PEG. Despite advanced HIV infection, serum biochemical parameters of nutritional status remained stable at 3-8 wk after PEG. Nine HIV patients suffered 10 complications during a mean follow-up of 111 +/- 147 (SD) days. The HIV patients had six (43%) minor complications, whereas the controls had four (19%) minor complications (not significantly different, Fisher's exact test). Minor complications in the HIV patients included transient tenderness and erythema at the PEG site in four, transient pyrexia without a source in one, and transient ileus in one. The HIV patients had four (29%) significant complications, compared to none in the controls (p < 0.02, Fisher's exact test). Significant complications in the HIV seropositive patients included stomal cellulitis treated with intravenous antibiotics in three, and gastric bleeding requiring transfusion of one unit of packed erythrocytes in one. None of these complications were severe; all complications rapidly resolved with therapy. The high rate of wound infection following PEG in AIDS patients, like the previously reported high rate of postoperative wound infection in AIDS patients, is attributable to immunosuppression. During the study period, four HIV patients underwent surgical gastrostomy, with one complication of severe gastrointestinal bleeding. CONCLUSIONS: This work suggests that PEG is a useful and relatively safe method of providing long-term nutritional support in selected AIDS patients with a functional gastrointestinal tract. PMID- 8249975 TI - Bacterial infection in cirrhosis, with and without hepatocellular carcinoma. AB - OBJECTIVE: The extent and type of bacterial infection occurring with liver cirrhosis has remained unknown. Further, while hepatocellular carcinoma (HCC) is known to occur mainly in patients with liver cirrhosis, no report has yet investigated the incidence of bacterial infection in HCC. The purpose of the present study was to establish the prevalence of bacterial infection in patients with HCC. METHODS: We have retrospectively investigated all 1140 patients with hepatic cirrhosis and/or HCC for any incidence of bacterial infection. RESULTS: The incidence of bacterial infection was found to be 15.4%, in 740 patients with HCC. This was approximately equal to the incidence of bacterial infection in 400 patients with cirrhosis without HCC, which was found to be 15.3%. When the severity of cirrhosis was graded according to Child-Pugh classification, the incidences of bacterial infection in Child-Pugh class A, class B, and class C were 3.3%, 11.1%, and 31.2%, respectively, in cirrhosis, and 2.3%, 9.1% and 25.6% in HCC. The incidence of bacterial infection increased with the severity of cirrhosis and severe bacterial infections occurred in Child-Pugh class B and C patients. CONCLUSION: The data suggest that the susceptibility of HCC patients to bacterial infection is mainly related to the underlying cirrhosis and not to the HCC. PMID- 8249976 TI - Magnetic resonance cholangiography for evaluation of obstructive jaundice. AB - OBJECTIVES: Magnetic resonance (MR) cholangiography is a newly developed adjunctive tool for noninvasive evaluation of patients with obstructive jaundice. We discuss whether MR cholangiography can provide useful information about the biliary tract, compared with other diagnostic modalities. METHODS: MR cholangiographies of 20 patients with obstructive jaundice were reviewed to evaluate the site of obstruction, the cause of obstruction, and the degree of stricture. RESULTS: The precise location of an obstructing lesion and complete display of the dilated biliary tree just proximal to the obstruction were provided in all 20 patients by MR cholangiography the same as in direct cholangiography. MR cholangiography could completely distinguish common duct stones from malignancies, whereas accurate differentiation among the malignancies was impossible. In six patients, MR cholangiography was not able to help in differentiating between high-grade stenosis and complete occlusion. CONCLUSION: Although precise cholangiographic estimation of the morphology and extent of the malignant disease by MR cholangiography was inferior to direct cholangiography, MR cholangiography is easily performed for a short duration and is a noninvasive diagnostic modality for assessing the outline of the anatomy of the dilated biliary tree. Therefore, we recommend promptly performing MR cholangiography if obstructive jaundice is suspected on clinical grounds. PMID- 8249977 TI - A syndrome of cirrhosis, achlorhydria, small intestinal bacterial overgrowth, and fat malabsorption. AB - OBJECTIVES: The purposes of this report were to examine whether or not a positive bile acid breath test in cirrhotic patient was associated with bacterial overgrowth in the upper small intestine, to verify that these bacteria have deconjugation ability, and, in addition, to elucidate whether or not changes in the gastric pH are related to bacterial overgrowth. METHODS: Twenty-seven patients with liver cirrhosis were tested by breath analysis technique using glycine-1-14C-labeled glycocholate. Jejunal fluids were aspirated through a double-lumen tube with a rubber cover on the tip. Anaerobes and aerobes were isolated and identified. After culturing, we used thin layer chromatography to determine whether each bacteria had the ability to deconjugate bile salts. RESULTS: Expired breath samples from seven of 27 patients with liver cirrhosis showed a marked increase of 14CO2-specific activity. Bacterial overgrowth was found in the jejunal fluid of these patients. Administration of chloramphenicol to the seven patients reduced 14CO2 specific activity significantly. The majority of the species identified deconjugated bile acids. Seventeen healthy control subjects showed no increase in CO2 excretion, and 16 of the 17 had no bacteria isolated from jejunal fluid. The relationship between 14CO2 specific activity of expired breath samples and gastric pH was also tested in the cirrhotic patients. There was a relatively good correlation (n = 27, r = 0.87, p < 0.05) between 14CO2 activity and gastric pH. CONCLUSION: We conclude that some patients with liver cirrhosis have bacterial overgrowth in the proximal small intestine that contains species that can deconjugate bile salts, and that the bacterial overgrowth is probably associated with the shift to alkaline pH in gastric juice. PMID- 8249978 TI - Partial regression of Barrett's esophagus--an inadequate endpoint. AB - A patient with Barrett's esophagus and high grade dysplasia is presented. He underwent a fundoplication and, subsequently, nearly three years of high-dose proton pump inhibition therapy. At the time of esophagogastrectomy, extensive squamous re-epithelialization was observed along with two histologic foci of intramucosal adenocarcinoma. Despite partial regression of Barrett's esophagus, high grade dysplasia present for more than 4 years had progressed to adenocarcinoma. Partial squamous repopulation of Barrett's esophagus is an inadequate endpoint of therapy. Complete regression is necessary to eliminate the risk of cancer. PMID- 8249979 TI - Hepatic infarction complicating a transjugular intrahepatic portosystemic shunt. AB - Transjugular intrahepatic portocaval shunt (TIPS) is being used with increasing frequency to treat variceal bleeding and portal hypertension. Like any other invasive vascular procedure, it has the potential for serious complications. We report a case of a woman who underwent a TIPS procedure for variceal bleeding due to hepatitis C and sustained a hepatic infarction as a complication of the procedure. We caution about the potential for catastrophic complications with this procedure, and recommend that it be performed only in centers with a liver transplant program nearby. PMID- 8249980 TI - Atypical presentation of inflammatory bowel disease in the elderly. AB - Despite multiple invasive diagnostic procedures including exploratory laparotomy and surgical resection, our patient's diagnosis remained an enigma. However, given the clinical scenario and the documented PG, a trial of steroids was warranted. The patient has fared well since her treatment, with resolution of all her symptoms. We feel confident that her disease process is most consistent with, and is most likely, Crohn's disease. There are several lessons to be learned from this case: 1) Inflammatory bowel disease can present at any age and belongs in a clinician's differential diagnosis of fever and diarrhea. 2) Failure to consider IBD in an elderly patient may lead to significant delay in diagnosis, and may expose the patient to unnecessary and sometimes dangerous intervention. 3) IBD in the elderly generally follows the same clinical patterns seen in younger patients. 4) Appropriate therapy can lead to prompt control or even resolution of the signs and symptoms of IBD. PMID- 8249981 TI - Cystic duct anatomy: an endoscopic perspective. AB - Knowledge of the junction of the cystic and common hepatic duct is essential for endoscopic management of biliary tract disease. The cystic and common hepatic ducts were evaluated retrospectively in cholangiograms obtained for a variety of indications in 524 persons. The cysticohepatic junction was adequately visualized in 70%. Medial junctions were noted in 18% and a spiral configuration in 32%, both more common than reported. An 11% occurrence of parallel duct systems was less frequent than expected. In 10% of patients, the cystic duct entered the hepatic duct in the distal third of the extrahepatic biliary tree. The importance of understanding this anatomy is illustrated with selected cases of therapeutic biliary endoscopy and laparoscopic cholecystectomy complications. Suggestions are made for improving performance in this area. PMID- 8249982 TI - Asymptomatic primary biliary cirrhosis in HTLV-I-associated myelopathy. AB - A 54-yr-old woman was admitted for complaints of slowly progressive gait and bladder disturbance, and HTLV-I-associated myelopathy (HAM/TSP) was diagnosed by positive antibodies to HTLV-I, both in her serum and cerebrospinal fluid. In the course of a diagnostic evaluation, she was found to have asymptomatic primary biliary cirrhosis. In immunohistochemistry, a fair number of her hepatocyte nuclei stained strongly positive for HTLV-I core protein, suggesting that HTLV-I may infect hepatocytes. The present case demonstrates that the spectrum of HTLV-I infection may be broad, and suggests the relationship between HTLV-I-associated myelopathy and primary biliary cirrhosis by autoimmune process. PMID- 8249983 TI - Acyclovir-associated colitis. PMID- 8249984 TI - Spontaneous healing of a cytomegalovirus esophageal ulceration in a patient with acquired immunodeficiency syndrome. PMID- 8249985 TI - Unusual migratory complication of a gastrostomy tube retrieved by percutaneous needle puncture. PMID- 8249986 TI - Intrahepatic spleen presenting as a mass lesion. PMID- 8249987 TI - Treatment of a giant biloma and bile leak by ERCP stent placement. PMID- 8249988 TI - Hepatic fascioliasis of long-term evolution: diagnosis by ERCP. PMID- 8249989 TI - Ascaris lumbricoides resulting in acute cholecystitis and pancreatitis in the Midwest. PMID- 8249990 TI - Benign bifurcation of the pancreatic duct presenting as a pancreatic mass on CT scan. PMID- 8249991 TI - Nutrition and colonic polyps: are we what we eat? PMID- 8249992 TI - Chronic idiopathic diarrhea: is there a happy ending to the story? PMID- 8249993 TI - Non-ulcer dyspepsia: seeing through the haze? PMID- 8249994 TI - Gallstones and colon cancer. PMID- 8249995 TI - A multicenter, randomized, controlled trial to evaluate the effect of prophylactic octreotide on ERCP-induced pancreatitis. PMID- 8249996 TI - Liver biopsy in patients with acquired immunodeficiency syndrome. PMID- 8249997 TI - Variceal band ligation and infection. PMID- 8249998 TI - Laci LeBeau syndrome. PMID- 8249999 TI - Crohn's disease presenting as pyogenic liver abscess. PMID- 8250000 TI - Hemobilia in a case of Bernard-Soulier syndrome. PMID- 8250001 TI - Mesenteric venous thrombosis with protein S deficiency. PMID- 8250002 TI - Echographic diagnosis of bowel intussusception. PMID- 8250003 TI - T- and B-cell markers in colonic nodular lymphoid hyperplasia in HIV infection. PMID- 8250004 TI - Idiopathic hypertrophic subaortic stenosis, aortic stenosis, and angiodysplasia. PMID- 8250005 TI - Characteristics of zinc binding to human red blood cell membranes. AB - The objective of the present study was to standardize the analysis of zinc binding on human red blood cell (RBC) membranes in 20 normal adults. The displacement studies revealed that at the maximal stable zinc concentration tested (600 microM), 57% (mean) of the bound 65Zn was displaced and to displace half maximal 65Zn, the stable zinc concentration was 300 microM. Scatchard plots revealed two classes of binding sites for zinc on RBC membranes: one with higher affinity, Kd = 1.20 x 10(-5) M (site I), and the other with lower affinity, Kd = 2.77 x 10(-4) M (site II). Binding sites occupancy was 97% means and 58.5% means for sites I and II, respectively. The displacement was affected by temperature, membrane protein concentration, freezing, thawing, and dialysis. Other metal cations, including Co++, Fe++, and Mn++, had very little effect on 65Zn displacement, in contrast copper displaced 65Zn from its binding sites on RBC membranes. Zinc binding to RBC membranes was rapid and readily reversible in a dynamic equilibrium with its binding sites. It is anticipated that this method will be applicable to studies of a wide variety of diseases specifically related to zinc metabolism in humans as well as in animals. PMID- 8250006 TI - Quantitative defect of glycoprotein Ib in severe cirrhotic patients. AB - A decrease of platelet agglutination induced by ristocetin has been described in cirrhotic patients. In order to investigate the relationship of such phenomenon with a putative defect on platelet membrane glycoprotein Ib, we have studied the quantitative and functional status of Gp Ib in eleven severe alcoholic cirrhotic patients, and the ability of their formalin-fixed platelets to agglutinate in the presence of normal plasma plus ristocetin. Interestingly, we found a significant decrease of immunoreactive GP Ib molecules (9,978 +/- 1,534 vs. 17,064 +/- 404 molecules per platelet) and ristocetin-dependent binding of vWF (9,113 +/- 1,338 vs. 13,992 +/- 1,968 molecules per platelet) (P < 0.01) in comparison to the levels found in a control group of healthy subjects. Immunoblotting analysis of platelet lysates confirmed the reduction of GP Ib level in cirrhotic patients, but showed no modification on the precipitation pattern of this glycoprotein. The ristocetin-induced platelet agglutination (light transmission %) was also significantly lower in patients with cirrhosis (48 +/- 7.6 vs. 92 +/- 3.6, P < 0.01), and correlated with the binding of normal vWF (r = 0.863, P = 0.0013). Patients with bleeding times longer than 7 min and/or clinical history of bleeding episodes showed the lowest values of platelet agglutination and GP Ib level. In conclusion, our present results indicate that liver cirrhosis is associated with a relevant decrease of functional GP Ib molecules on the platelet surface. This reduction might be related to the prolongation of bleeding time and to the bleeding diathesis observed in cirrhotic patients. PMID- 8250007 TI - HTLV-I associated adult T-cell leukaemia/lymphoma in Hong Kong. PMID- 8250008 TI - Direct evidence for systemic fibrinogenolysis in patients with acquired alpha 2 plasmin inhibitor deficiency. AB - To examine whether or not acquired alpha 2-plasmin inhibitor deficiency is associated with systemic fibrinogenolysis, we analyzed the fibrin and fibrinogen degradation products in eight patients with this condition in various disease states. The underlying disease was gastric cancer in three patients, metastatic prostatic cancer in two, acute promyelocytic leukemia in two, and abdominal aortic aneurysm in one patient. In all eight patients, the alpha 2-plasmin inhibitor level was reduced to less than 50% of normal, and plasmin-alpha 2 plasmin inhibitor complex levels were increased. Immunoblotting of serum using an antifibrinogen antibody detected a 250 kDa protein (corresponding to fragments X or DY) in all eight patients. Fragment Y and D monomer were detected in seven of the eight patients, indicating the occurrence of systemic fibrinogenolysis. However, they were not detected in one patient with metastatic prostatic cancer. To determine whether or not fibrinogen degradation was also occurring in the patient without fragment Y, we characterized the 250 kDa protein in all eight patients. The protein was found to be fragment X in the metastatic prostatic cancer patient without fragment Y, while it was fragment DY in the other seven patients. Thus, systemic fibrinogenolysis was present in all eight patients. In the two patients with metastatic prostatic cancer, the level of alpha 2-plasmin inhibitor gradually increased with the reduction of tumor size by treatment. Fragment X, fragment Y, and D monomer were not detected when the alpha 2-plasmin inhibitor level exceeded 60% of normal in both patients. In the other six patients fragment Y and D monomer also disappeared when the alpha 2-plasmin inhibitor level exceeded 60% of normal. These findings suggest that systemic fibrinogenolysis only occurs when the plasma levels of alpha 2-plasmin inhibitor falls below 60% of normal due to activation of the fibrinolytic system by various pathological conditions. PMID- 8250009 TI - Detection of tumor necrosis factor-alpha in bone marrow plasma and peripheral blood plasma from patients with aplastic anemia. AB - Plasma levels of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) were determined in healthy individuals and patients with aplastic anemia (ApAn). IFN-gamma was not detected in normal peripheral blood plasma (PBP) or bone marrow plasma (BMP) and was present in PBP from only 2 of 22 patients and in BMP from 1 of 14 patients and the levels were low (< 1.5 U/ml). Elevated levels of TNF-alpha were present in BMP and PBP from patients but not in control (healthy donor) PBP and BMP. Eleven of twenty-four patients had elevated levels of TNF-alpha in their PBP and 6 of 13 patients had detectable levels of TNF-alpha in their BMP. Only one of the 14 healthy control donors had detectable TNF-alpha and the level was very low (7 pg/ml), while 13 of the 27 ApAn patients had detectable TNF-alpha (P = .009, chi-square test). Not surprisingly, the centers of the distributions of TNF-alpha concentrations of the controls and ApAn patients differed significantly (P < .017 for control and patient PBP and P < .056 for control and patient BMP, Wilcoxon rank-sum test). Spontaneous production of IFN-gamma and TNF-alpha by cultured bone marrow mononuclear cells was observed in four of seven patients but not in the six healthy controls (P = 0.026). Spontaneous production of IFN-gamma and TNF-alpha by cultured peripheral blood mononuclear cells from patients and controls was however similar. Phytohemagglutinin (PHA)-induced production of IFN-gamma and TNF-alpha by cultured mononuclear cells did not differ significantly between ApAn patients and normal controls. The significance of overproduction of TNF-alpha in the pathophysiology of ApAn is discussed. PMID- 8250010 TI - Splenic lymphoma with circulating villous lymphocytes: report of seven cases and review of the literature. AB - Splenic lymphoma with villous lymphocytes (SLVL) is a relatively new entity with only a few reports published. We report seven cases of SLVL with detailed clinicopathologic and comprehensive immunophenotypic studies to further characterize this lymphoma, which is frequently confused with hairy cell leukemia and other low-grade B-cell lymphoid neoplasms. The diagnostic criteria we used include 1) prominent splenomegaly, 2) insignificant or no lymphadenopathy, 3) lymphocytosis without leukopenia, 4) presence of circulating villous lymphocytes, 5) characteristic cytologic and histologic features, and 6) specific phenotypic and cytochemical findings. Our studies show that SLVL does not represent a pure entity but rather a morphologically heterogeneous group of low-grade lymphomas with various cytologic and histologic features. Although immunophenotyping is helpful in differential diagnosis, multiparameter studies are necessary to confirm the diagnosis. In our series, only two patients died of SLVL, who probably developed transformation to a higher-grade lymphoma. PMID- 8250011 TI - Dyshematopoiesis in combined immune deficiency with congenital neutropenia. AB - This report describes a patient with combined immune deficiency associated with congenital neutropenia (CID/CN) and reports a partial characterization of his hematopoietic abnormalities. The CID/CN syndrome described is characterized by neutropenia and by deficiencies in B-lymphoid and T-lymphoid cell number and function. Red cell and platelet counts were normal. In vitro assays indicate that the myeloid lineage was developmentally arrested at the level of the committed monocyte/granulocyte progenitor (CFU-GM), while precursors to the CFU-GM progenitor were normal. In vitro studies showed that the defect in myeloid development was not corrected with G-CSF or GM-CSF. However, combinations of cytokines present in conditioned media from the T-cell lines MO or C5MJ, or defined multiple cytokine combinations containing IL-1, IL-3, GM-CSF, kit ligand, IL-6, and IL-9, restored myelopoiesis in-vitro. In contrast, C5MJ-conditioned media did not correct deficiencies in immune function in the patient's lymphocytes and accessory cells. No abnormalities in the production of G-CSF, GM CSF, M-CSF, or IL-1 from the patient could be identified to account for the defects in myelopoiesis orimmune function. PMID- 8250012 TI - Autoimmune-thrombocytopenia and SLE in a patient with 5q-anomaly and deletion of the c-fms oncogene. AB - Autoimmune-thrombocytopenia was the striking feature in a patient with typical clinical symptoms of systemic lupus erythematosus (SLE), complement C4 deficiency, and positive lupus serology. However, myelodysplasia was found in the bone marrow and chromosome analysis revealed a deletion of the long arm of chromosome 5 (5q-anomaly), which was confirmed by a hemizygosity for the c-fms oncogene (CSF-1-receptor) on Southern blot. Autoimmune phenomena reported in conjunction with myelodysplastic syndromes (MDS), e.g., an elevation of antinuclear antibodies, are usually regarded as nonspecific. This case report suggests that SLE can occur in patients with MDS and that a concomitant autoimmune-thrombocytopenia may mask the typical signs of the 5q- syndrome. PMID- 8250013 TI - Laboratory monitoring of warfarin therapy in Utah. AB - Accurate laboratory monitoring of oral anticoagulation has been emphasized as an important factor in providing safe and effective therapy for patients with thromboembolism. However, recent reports indicate that coagulation laboratories may not be providing optimal clinical information to clinicians who treat these patients. We surveyed all hospital coagulation laboratories in Utah to determine their format for reporting prothrombin time results in patients receiving oral anticoagulants. We found that less than 50% of laboratories used the reliable reporting format, i.e., the International Normalized Ratio (INR), and that many of the laboratories using the INR format may be reporting incorrect values. Our survey also found a significant lack of interest by physicians in requesting that their laboratories adopt reliable reporting methods. These results indicate a substantial lack of understanding by laboratories and clinicians of the importance of using reliable methods to monitor oral anticoagulation. Significant educational efforts will be required to correct this problem. PMID- 8250014 TI - Production of parathyroid hormone-related protein by cultured human myeloma cells. AB - We examined whether or not parathyroid hormone-related protein (PTH-rP) is produced in cultured human myeloma cells. PTH-rP protein was detected in the cell extract of one (U266) of 6 myeloma cell lines examined by radioimmunoassay. Also we demonstrated the expression of PTH-rP mRNA in all of the 6 lines. These findings indicate that PTH-rP may be one of the factors responsible for the bone destruction or hypercalcemia in multiple myeloma. PMID- 8250015 TI - Quantities of alpha Q chain variants in heterozygotes with and without a concomitant beta-thalassemia trait. AB - We have analyzed the quantities of alpha x chain-containing hemoglobins (alpha 2 x beta 2 and alpha 2 x delta 2) in 14 heterozygotes for Hb Q-India [alpha 64(E13)Asp-->His] or Hb Q-Thailand [alpha 74(EF3)Asp-->His]; both amino acid replacements are the result of mutations in the alpha 1-globin gene. Five of these persons (three with Hb Q-India and two with Hb Q-Thailand) had an additional beta(0)-thalassemia heterozygosity. The average quantities for Hb Q + Hb Q2 in the four groups were 17.2% (alpha alpha Q/alpha alpha; beta A/beta A), 9.5% (alpha alpha Q/alpha alpha; beta A/beta(0) Th), 26.8% (-alpha Q/alpha alpha; beta A/beta A), and 16.95% (-alpha Q/alpha alpha; beta A/beta(0) Th). These variations can best be explained by a posttranslational control mechanism; an imbalance in the alpha A, alpha Q, and beta A chain ratio will favor the alpha 2 Q beta 2 formation when an alpha-thalassemia is present and will reduce its formation in the presence of a beta-thalassemia heterozygosity. PMID- 8250016 TI - Acute lymphoblastic leukemia in Fanconi's anemia. PMID- 8250017 TI - Standard Ph chromosome, t(9;22)(q34;q11), as an additional change in a patient with acute myelomonocytic leukemia (M4Eo) associated with inv(16)(p13q22) PMID- 8250018 TI - Beta S-gene-cluster haplotypes in sickle cell anemia patients from two regions of Brazil. PMID- 8250019 TI - Burkitt's lymphoma presenting as ophthalmoplegia in an elderly male. PMID- 8250020 TI - Intracranial hemorrhage in an adult with factor-VII deficiency. PMID- 8250021 TI - High-dose human immunoglobulins in thrombotic thrombocytopenic purpura. PMID- 8250022 TI - Renal replacement therapies in the elderly: Part 1. Hemodialysis and chronic peritoneal dialysis. AB - Recent demographic data from the United States and Europe demonstrate that the dialysis population is growing progressively older. In the United States the median age at onset of end-stage renal disease (ESRD) had reached 61 years in 1990, and the fraction of ESRD patients older than 65 years will approach 60% by the year 2000. The primary treatment of geriatric ESRD is center hemodialysis (82% of elderly patients). Chronic peritoneal dialysis in this age group is limited in the United States to less than 10%. Only 2.7% of elderly patients have a functioning transplant. Despite their complex medical and psychosocial conditions, survival and rehabilitation are acceptable in the elderly dialysis patient and these patients tend to be more complaint. Five-year dialysis patient survival rates in the United States are 19% and 10%, respectively, for the 65 to 74 and 75 to 84 age groups compared with 32% and 19%, respectively, for the same age groups in Europe. Five-year survival, particularly in the elderly, is even higher in Japan. Several factors favor the delivery of low doses of hemodialysis in the elderly and discontinuing dialysis is more common in the elderly. Continuous ambulatory peritoneal dialysis is a satisfactory alternative treatment modality for geriatric ESRD. Most studies confirm that survival of elderly patients on continuous ambulatory peritoneal dialysis and hemodialysis is similar. The 1991 United States Renal Data System report showing higher mortality among diabetic continuous ambulatory peritoneal dialysis patients than among hemodialysis patients may reflect selection of such patients, who may have more co-morbid conditions. In special cases, continuous cyclic peritoneal dialysis can be an alternative treatment for elderly ESRD patients. PMID- 8250023 TI - Renal disease in adolescents with type I diabetes mellitus: a report of the Southwest Pediatric Nephrology Study Group. AB - Although 30% to 40% of patients with type I diabetes mellitus develop diabetic nephropathy, the usual signs of clinical nephropathy are often thought to be delayed until adulthood. We studied 13 adolescents with type I diabetes mellitus for 5 to 14 years who had renal biopsies completed because of clinical problems, including proteinuria, hematuria, or hematuria plus proteinuria. Changes typical of diabetes were seen by light and immunofluorescence microscopy; evidence of other renal diseases was not noted. On electron microscopy evaluation, glomerular basement membrane width was increased in 11 patients. In seven patients, mesangial volume was elevated above the normal range. Peripheral capillary filtration surface density was diminished below the normal range in five patients. Thus, several of these adolescents had severe glomerular lesions that were indicative of overt diabetic nephropathy. Within 2 to 3 years after biopsy, at least two patients were dialysis dependent. Thus, the adolescent diabetic patient with a relatively short duration of diabetes may be developing progressive diabetic renal lesions, and the clinical signs and symptoms at presentation may not be those typically seen in diabetic nephropathy. PMID- 8250024 TI - The association of non-insulin-dependent diabetes mellitus and hypertension with urinary excretion of albumin and transferrin. AB - Abnormal urinary excretion of albumin is a nonspecific sign of nephropathy, commonly occurring in persons with hypertension as well as diabetes. Transferrin, rather than albumin, is more readily excreted by the kidney in those with diabetes compared with those with hypertension alone. One hundred eighty non insulin-dependent diabetic mellitus patients were age, race, and sex matched to 90 nondiabetic control subjects who had mild to moderate hypertension. Overnight urine collections were analyzed by immunoturbidity for albumin and transferrin. The average duration of hypertension was 11 years among controls. Mean blood pressures were 134/79 mm Hg for diabetic subjects and 145/87 mm Hg for controls (P < 0.001). Diabetic patients had higher mean excretion rates of albumin (128 micrograms/min v 49 micrograms/min; P = 0.04) and transferrin (7.3 micrograms/min v 0.9 microgram/min; P = 0.0001) and higher excretion ratios of albumin (0.179 g/g creatinine v 0.069 g/g creatinine; P = 0.02) and transferrin (0.0065 g/g creatinine v 0.0013 g/g creatinine; P < 0.001) than hypertensive controls. Ratios of transferrin to albumin excretion for those with diabetes and hypertension greatly exceeded expected ratios for those with hypertension or diabetes alone if the effects of these disorders were additive. While diabetic patients were twice as likely as controls to have abnormal albumin excretion ratios (P = 0.01), they were three times as likely to have elevated transferrin excretion ratios (P = 0.0001), even though the diabetic group was half as likely as the controls to have systolic blood pressure > or = 160 mm Hg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250025 TI - The quantitative relationship between treated blood pressure and progression of diabetic renal disease. AB - Antihypertensive therapy reduces the rate at which glomerular filtration rate (GFR) declines (delta GFR) in diabetic nephropathy; however, the optimal blood pressure is unknown. The quantitative relationship between treated blood pressure and delta GFR was analyzed retrospectively in 59 patients with established diabetic nephropathy and treated hypertension using weighted univariate and weighted multivariate regression. The GFR was calculated using the Cockcroft and Gault formula. More rapid GFR loss correlated most strongly with higher diastolic blood pressures (r = 0.70; P < 0.0001); for each millimeter of mercury of diastolic blood pressure, the GFR decreased by 0.69 mL/min/yr. This relationship remained present if those individuals with diastolic pressures greater than 90 mm Hg were eliminated from the study (r = 0.50; P < 0.001). The correlation for systolic blood pressure was weaker (r = 0.30; P < 0.05) and explained completely by covariance between systolic and diastolic blood pressures. The correlation for mean blood pressure (r = 0.59; P < 0.0001) fell between the correlations for diastolic and systolic blood pressures. Proteinuria, serum albumin concentration, and serum cholesterol concentration also correlated with delta GFR. In multivariate analysis, neither these indices of disease severity nor the initial GFR explained the correlation between delta GFR and diastolic blood pressure. Age, sex, race, type of diabetes, and percentage of glycosylated hemoglobin did not correlate with delta GFR. PMID- 8250026 TI - An ultrasound renal cyst prevalence survey: specificity data for inherited renal cystic diseases. AB - This study documents the prevalence of simple renal cysts at various ages to produce a table of age-specific specificity values. The specificity of diagnosis for inherited renal cystic conditions depends primarily on the prevalence of simple renal cysts in disease-free subjects. To provide specificity data useful for the ultrasound assessment of individuals at genetic risk of having a renal cyst-forming condition, a prospective ultrasound study was performed. The urinary tract was examined in detail in 729 individuals referred for investigation of symptoms unrelated to the urinary tract and who had normal renal function. The prevalence of individuals with at least one renal cyst was 0% in those aged 15 to 29 years, 1.7% in those aged 30 to 49 years, 11.5% in those aged 50 to 70 years, and 22.1% in those aged 70 years and above. The prevalence of bilateral renal cysts (at least one cyst in each kidney) was 1% in those aged 30 to 49 years, 4% in those aged 50 to 70 years, and 9% in those aged 70 years and above. A table is provided that allows specificity estimates for differing diagnostic criteria together with 95% confidence limits for each estimate. In the younger age groups, bilateral renal cysts, particularly bilateral cysts with one or both kidneys having more than one cyst, are relatively uncommon. A reduced threshold for suspecting inherited renal cystic disease in younger individuals at high genetic risk will continue to have high specificity while improving the sensitivity of ultrasound diagnosis. PMID- 8250027 TI - Elevated lipoprotein (a) levels in primary nephrotic syndrome. AB - Elevated plasma levels of total cholesterol and increase in the hepatic synthesis of some apo B-containing lipoproteins have been noted in the nephrotic syndrome. Apoprotein (a), the apolipoprotein distinguishing lipoprotein (a) [Lp(a)] from low-density lipoprotein, is equally of hepatic origin, and Lp(a) recently has been shown to possess both atherogenic and thrombogenic activities. However, little is known of Lp(a) levels in nephrotic patients. We measured plasma Lp(a) concentrations in 11 patients with primary nephrotic syndrome in the absence of hematuria, hypertension, and renal insufficiency. Histologic lesions were minimal change disease in five cases, membranous glomerulopathy in four cases, and focal and segmental glomerulosclerosis in two cases. Mean levels of Lp(a) (98 +/- 92 mg/dL [mean +/- SD]) were markedly elevated in the nephrotic patients as compared with the controls (14 +/- 13 mg/dL). No correlation was noted between plasma Lp(a) and proteinuria, albuminemia, total cholesterolemia, low-density lipoprotein cholesterol, apoprotein B100, or plasminogen. Furthermore, there was no correlation between Lp(a) levels and apoprotein (a) isoform size. In four patients, the level of Lp(a) decreased approximately fourfold after remission of the nephrotic syndrome under corticosteroid treatment. Our observation that Lp(a) levels are elevated in the nephrotic syndrome is consistent with the hypothesis that these patients may be at an increased risk of cardiovascular and thrombotic complications. PMID- 8250028 TI - Fingernail creatinine as a predictor of prior renal function. AB - When a patient presents with renal failure, it is often difficult to ascertain whether the individual is suffering from acute or chronic renal failure. Fingernail creatinine might help to differentiate between the two. To test this possibility, the relationship between the fingernail creatinine and the serum creatinine obtained concurrently or 1 or more months previously was examined in 22 normal adults, nine patients with acute renal failure, seven patients with chronic renal failure not undergoing dialysis, 16 maintenance hemodialysis patients, and 33 patients with a functioning renal transplant who had been transplanted 0.2 to 3.9 months (n = 21) or 4.3 to 33 months previously (n = 12). Fingernail creatinine was significantly greater than normal in the patients with chronic renal failure, patients undergoing maintenance hemodialysis, and patients who had a functioning renal transplant implanted 0.2 to 3.9 months previously. In contrast, fingernail creatinine was not different from normal in the patients with acute renal failure and in patients who had a functioning renal transplant placed 4.3 to 33 months previously. In the maintenance hemodialysis patients, fingernail creatinine showed the strongest correlation with the serum creatinine obtained between 5 and 10 months previously, with the highest correlation at 9 months. In the renal transplant recipients, the fingernail creatinine decreased to normal or near normal values within approximately 90 to 120 days after transplantation. These findings indicate that fingernail creatinine may reflect the serum creatinine values several months previously. The fingernail creatinine may help to identify whether patients have recent onset as compared with longstanding renal failure. PMID- 8250029 TI - The relationship between plasma and dialysate lipoproteins and apoproteins in patients treated by continuous ambulatory peritoneal dialysis. AB - End-stage renal disease is frequently associated with lipoprotein abnormalities, manifested primarily by elevated very low-density lipoprotein levels combined with a decrease in high-density lipoprotein levels. These lipoprotein disturbances are further exacerbated in continuous ambulatory peritoneal dialysis. We examined the lipoprotein and apolipoprotein profiles in the blood and dialysate effluents of eight normolipidemic and five hypertriglyceridemic patients with end-stage renal failure treated with continuous ambulatory peritoneal dialysis. The normolipidemic patients were found to have significantly greater losses, as expressed by the fractional catabolic rates through the dialysate, for protein, total cholesterol, and very low-density lipoprotein cholesterol. These results suggest that the hypertriglyceridemia associated with continuous ambulatory peritoneal dialysis may be mitigated in some patients by the excessive loss of very low-density lipoprotein, or some other plasma constituent, into the dialysate effluent. PMID- 8250030 TI - Influence of race and gender on related donor renal transplantation rates. AB - Racial differences in kidney transplantation have received recent attention both in the medical community and in the general public. Most efforts to improve renal transplantation among minority groups have been directed toward cadaveric donation and transplantation. Since evaluation of the comparative trends by race of living related donor (LRD) kidney transplantation have been lacking, we examined trends of LRD transplantation from 1983 through 1990 using national data from the US Renal Data System. The total number of LRDs in blacks did not change during the 8-year period between 1983 and 1990 (198 in 1983 and 197 in 1990). During this same period, the total number of LRDs in whites increased by 11% (1,390 in 1983 and 1,548 in 1990). Rates of LRD transplantation per nontransplanted dialysis patients were consistently lower in blacks and females compared with whites and males, respectively. White males have a fivefold higher rate of LRD transplantation than black males, whereas white females have a fourfold higher rate then black females. When intraracial gender differences were examined, black males were transplanted with LRD kidneys at a rate 20% higher than black females. This difference was present between 1983 and 1989, but was nonexistent in 1990. Among whites, males also had a higher rate of LRD transplantation than females, which gradually decreased from 34% in 1983 to 20% in 1990. In view of the ever-increasing demand for cadaver organs, additional effort in the medical community and society toward increasing LRD transplantation rates represents a more promising approach to increasing organ donation in all groups than a single focus on cadaveric donation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250031 TI - Renal responses to acute angiotensin II inhibition and administered angiotensin II in the aging, conscious, chronically catheterized rat. AB - Studies of conscious, chronically catheterized, young (3 to 5 months of age) and old (19 to 22 months of age) male Sprague Dawley rats in the baseline state showed that glomerular filtration rate (factored for body weight) was lower and urine flow higher in old compared with young rats. Acute blockade of endogenous angiotensin II (AII) with either converting enzyme inhibitor (CEI) or Losartan (Dupont Merck, Wilmington, DE) caused small variable decreases in blood pressure in both age groups and produced a significant renal vasodilation with increases in renal plasma flow in the older rats. In separate studies using low-dose AII infusion (5 ng/kg body weight/min), an increase in filtration fraction was the only effect seen in both young and old rats with an intact renin/AII system; no effect was seen with CEI. During high-dose AII (20 ng/kg body weight/min), significant and similar increases were seen in blood pressure and renal vascular resistance in old and young rats, and the patterns of blood pressure and renal hemodynamic responses were similar with and without acute CEI. A natriuretic and diuretic response to high-dose AII was seen in young rats with intact endogenous renin/AII, whereas old rats were completely refractory to this action of AII. Against a background of acute CEI, no natriuretic/diuretic response to high-dose AII was seen in either age group. Thus, in the baseline state, renal hemodynamics in the old kidney are controlled by endogenous AII since CEI and Losartan produce renal vasodilation in old but not young rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250032 TI - Renal tissue angiotensin II: response to infusions of angiotensin I and an angiotensin-converting enzyme inhibitor. AB - A procedure for the collection, processing, extraction, separation, and measurement of renal tissue angiotensin II [ANG-(1-8)] is described. The arterial plasma and renal tissue concentrations of ANG-(1-8) were examined in anesthetized Sprague-Dawley male rats (10 to 12 weeks of age) given an intravenous saline infusion (group 1; n = 6), an intravenous bolus infusion of 0.5 microgram angiotensin I (group 2; n = 6), or an intravenous bolus of an angiotensin converting enzyme inhibitor (group 3; n = 6). Plasma and renal tissue were collected at the peak mean arterial pressure (MAP) response. The mean (+/- SEM) concentration of ANG-(1-8) in the group given the saline vehicle was 12 +/- 2 fmol/mL in the plasma and 2.4 +/- 0.3 pmol/g in the renal tissue. The angiotensin I bolus significantly increased MAP by 40%. Following the angiotensin I infusion, at the time of peak MAP response, the concentration of the circulating and renal tissue ANG-(1-8) were 12-fold and twofold higher, respectively, compared with the saline vehicle. The angiotensin-converting enzyme inhibitor significantly decreased MAP by 10% and decreased the circulating and renal tissue ANG-(1-8) levels by 75% and 62.5%, respectively, compared with the saline vehicle. The rapid conversion of exogenous angiotensin I to ANG-(1-8) in renal tissue provides direct evidence that renal tissue can generate de novo ANG-(1-8) from a circulating precursor. Furthermore, plasma and renal tissue respond similarly to acute angiotensin-converting enzyme inhibition. PMID- 8250033 TI - Regulation of epidermal growth factor receptor expression and growth by protein kinase C and retinoic acid in LLC-PK1 cells. AB - The importance of receptor expression and protein kinase C to epidermal growth factor (EGF)-induced cell proliferation was studied in LLC-PK1 kidney cells. These cells have both high- and low-affinity binding sites for EGF. Neither transforming growth factor-beta nor tumor necrosis factor altered EGF receptor expression. On the other hand, retinoic acid induced a concentration-dependent increase in EGF binding that was maximal at 1 mumol/L. One micromolar of retinoic acid increased EGF binding from 0.38 +/- 0.01 fmol/10(6) cells in controls to 1.10 +/- 0.03 fmol/10(6) in treated cells at 18 hours (n = 8, P < 0.001). The increase in binding was the result of an increase in the Bmax of the high affinity receptor. The upregulation of the EGF receptor induced by retinoic acid was associated with enhanced EGF-induced growth promotion. A 45-minute incubation of cells with phorbol 12-myristate 13-acetate caused a concentration-dependent decrease in EGF binding that was prevented by a 40-hour, 2 mumol/L pre-exposure to phorbol 12-myristate 13-acetate; 10(-8) mol/L EGF also caused a downregulation of the EGF receptor that was not prevented by phorbol 12-myristate 13-acetate or retinoic acid. Downregulation of protein kinase C did not interfere in the capacity of EGF to induce growth in these cells. These studies demonstrate that EGF receptor upregulation plays an important role in the control of EGF-induced cell growth. Protein kinase C regulates EGF binding in these cells; however, it is not necessary for EGF-induced growth promotion or receptor downregulation. PMID- 8250034 TI - Acute renal failure due to rhabdomyolysis associated with the extreme lithotomy position. AB - A patient who developed acute renal failure secondary to rhabdomyolysis associated with the use of the extreme lithotomy position for 6 hours during radical perineal prostatectomy is described. It appears that muscle ischemia due to compression of the lumbar and pelvic muscles resulted in muscle injury. Intense muscle uptake of technetium 99m methylene diphosphonate assisted in localizing the muscles involved and ascertaining the extent of the injury. Review of the literature disclosed seven other patients with a similar association. All patients complained of muscle pain shortly after recovery from anesthesia. Early recognition and aggressive treatment with intravenous fluids may prevent the development of acute renal failure. PMID- 8250035 TI - Reflex anuria and uremia from unilateral ureteral obstruction. AB - A 59-year-old man had anuria of 32 hours' duration that was associated with unilateral calculous ureteral obstruction. Retrograde pyelography showed a normal contralateral collecting system. Initial technetium pentetate (DTPA) scanning showed diminished uptake of radioactivity by the unobstructed kidney compared with the obstructed kidney, whereas a follow-up study, after relief of ureteral obstruction, showed a reversal of this pattern. Previously reported cases of "reflex anuria" are reviewed along with the relevant experimental literature. Vascular or ureteral spasm related in part to abnormality of the autonomic nervous system may underlie this rare entity. PMID- 8250036 TI - Primary focal segmental glomerulosclerosis: pathology, histological variants, and pathogenesis. PMID- 8250037 TI - Seven momentous years. PMID- 8250038 TI - Independent origin of identical beta cardiac myosin heavy-chain mutations in hypertrophic cardiomyopathy. AB - The origins of the beta cardiac myosin heavy-chain (MHC) gene missense mutations that cause familial hypertrophic cardiomyopathy (FHC) in 14 families have been evaluated. Of eight different mutations, four were present in single families, while four occurred in two or more families. To investigate the origins of the four shared mutations, we defined the beta cardiac MHC haplotypes of each of the mutation-bearing chromosomes by determining the alleles present at three intragenic polymorphic loci. Two of the mutations (Arg453Cys and Val606Met) have arisen independently in each of three families, being found on different chromosomal backgrounds. A third mutation (Gly584Arg) is associated with identical haplotypes in two families with Portuguese ancestors, suggesting a founder effect. Haplotype analysis was uninformative for the fourth mutation (Arg403Gln). Thus, FHC-causing mutations have arisen independently in at least 12 of the 14 families studied, suggesting that the majority have arisen relatively recently as new mutations. This finding predicts the prevalence of disease causing beta cardiac MHC mutations to be comparable in all population groups. PMID- 8250039 TI - Physical mapping by FISH of the DiGeorge critical region (DGCR): involvement of the region in familial cases. AB - We describe the relative ordering, by fluorescence in situ hybridization, of cosmid loci and translocation breakpoints in the DiGeorge syndrome (DGS) critical region of chromosome 22. This physical map enables us to define a large region, commonly deleted in a majority of affected patients, and the smallest deleted region which, when lost, is sufficient to produce DGS. In four instances, a similar large deleted region is observed in a familial context. In these pedigrees, the deletion is encountered in one parent with mild features of the disease. PMID- 8250040 TI - Localization of the Krabbe disease gene (GALC) on chromosome 14 by multipoint linkage analysis. AB - The gene responsible for Krabbe disease, an autosomal recessive disorder caused by deficiency of galactocerebrosidase (GALC), was localized by multipoint linkage analysis on chromosome 14. Eight mapped dinucleotide repeat polymorphisms were tested for linkage to GALC. Two-point linkage analysis demonstrated close linkage of GALC and D14S48, with Z = 13.69 at theta = 0. Multipoint analysis yielded strong support for this finding, with maximum likelihood for GALC located within 1 cM of D14S48. This analysis also identified markers that clearly flank the GALC locus, as the map order of D14S53-GALC-D14S45 is favored by odds greater than 10(6):1. Additional support for close linkage of GALC and D14S48 comes from the apparent linkage disequilibrium between these two loci in a consanguineous Druze community in Israel. These data localize GALC to 14q24.3-q32.1. PMID- 8250041 TI - Mapping of a gene for familial juvenile nephronophthisis: refining the map and defining flanking markers on chromosome 2. APN Study Group. AB - Familial juvenile nephronophthisis (NPH) is an autosomal recessive kidney disease that leads to end-stage renal failure in adolescence and is associated with the formation of cysts at the cortico-medullary junction of the kidneys. NPH is responsible for about 15% of end-stage renal disease in children, as shown by Kleinknecht and Habib. NPH in combination with autosomal recessive retinitis pigmentosa is known as the Senior-Loken syndrome (SLS) and exhibits renal pathology that is identical to NPH. We had excluded 40% of the human genome from linkage with a disease locus for NPH or SLS when Antignac et al. first demonstrated linkage for an NPH locus on chromosome 2. We present confirmation of linkage of an NPH locus to microsatellite markers on chromosome 2 in nine families with NPH. By linkage analysis with marker AFM262xb5 at locus D2S176, a maximum lod score of 5.05 at a theta max = .03 was obtained. In a large NPH family that yielded at D2S176 a maximum lod score of 2.66 at theta max = .0, markers AFM172xc3 and AFM016yc5, representing loci D2S135 and D2S110, respectively, were identified as flanking markers, thereby defining the interval for an NPH locus to a region of approximately 15 cM. Furthermore, the cytogenetic assignment of the NPH region was specified to 2p12-(2q13 or adjacent bands) by calculation of linkage between these flanking markers and markers with known unique cytogenetic assignment. The refined map may serve as a genetic framework for additional genetic and physical mapping of the region. PMID- 8250042 TI - A large, dominant pedigree of atrioventricular septal defect (AVSD): exclusion from the Down syndrome critical region on chromosome 21. AB - We describe a large pedigree of individuals with autosomal dominant atrioventricular septal defect (AVSD). The pedigree includes affected individuals and individuals who have transmitted the defect but are not clinically affected. AVSDs are a rare congenital heart malformation that occurs as only 2.8% of isolated cardiac lesions. They are the predominant heart defect in children with Down syndrome, making chromosome 21 a candidate for genes involved in atrioventricular septal development. We have carried out a linkage study in the pedigree by using 10 simple-sequence polymorphisms from chromosome 21. Multipoint linkage analysis gives lod scores of less than -2 for the region of trisomy 21 associated with heart defects, which excludes a locus within this region as the cause of the defect in this family. PMID- 8250043 TI - Clinical and molecular evaluation of four patients with partial duplications of the long arm of chromosome 18. AB - Four individuals with partial duplications of the long arm of chromosome 18 were analyzed at the clinical, cytogenetic, and molecular levels. Two of the individuals had duplications of the long arm from 18q21.1-qter because of inheritance of an unbalanced translocation. Both of these individuals displayed the clinical phenotype characteristic of Edwards syndrome. Two other patients had de novo interstitial duplications of 18q but did not have a clinical diagnosis of Edwards syndrome. The extent of the duplicated material in each patient was determined initially by using cytogenetic analysis and subsequently with more detailed comparisons of the duplicated regions by using molecular probes derived from a chromosome 18-specific lambda phage library. The results demonstrated that one of the de novo interstitial duplications that did not result in the Edwards syndrome phenotype had a more proximal breakpoint than that of the partial duplications of the two patients with features of Edwards syndrome. These results suggest that a single critical region for Edwards syndrome in the proximal portion of 18q is unlikely. PMID- 8250044 TI - An index marker map of chromosome 9 provides strong evidence for positive interference. AB - An index marker map of chromosome 9 has been constructed using the Centre d'Etude du Polymorphisme Humain reference pedigrees. The map comprises 26 markers, with a maximum intermarker interval of 13.1 cM and only two intervals > 10 cM. Placement of all but one marker into the map was achieved with > 10,000:1 odds. The sex equal length is 151 cM, with male length of 121 cM and female length of 185 cM. The map extends to within 2%-3% of physical length at the telomeres, and its coverage therefore is expected to be within 20-30 cM of full map length. The markers are all of the GT/CA repeat type and have average heterozygosity .77, with a range of .60-.89. The map shows both marked contraction of genetic distance relative to physical distance in the pericentromeric region and expansion in the telomeric regions. Genotypic data were carefully examined for errors by using the crossover routine of the program DATAMAN. Five new mutations were observed among 17,316 meiotic events examined. There were two double crossover events occurring within an interval of 0-10 cM, and another eight were observed within an interval of 10-20 cM. Many of these could be due to additional mutational events in which one parental allele converted to the other by either gene conversion or random strand slippage. When there was no correction for these possible mutational events, the number of crossovers displayed by the maternal and paternal chromosomes was significantly different (P < .001) from that predicted by the Poisson distribution, which would be expected in the absence of interference. In addition, the observed crossover distribution for paternally derived chromosomes was similar to that predicted from cytogenetic chiasma frequency observations. In all, the data strongly support the occurrence of strong positive interference on human chromosome 9 and suggest that flanking markers at an interval of < or = 20 cM are generally sufficient for disease gene inheritance predictions in presymptomatic genetic counseling by linkage analysis. PMID- 8250045 TI - Linkage analysis by two-dimensional DNA typing. AB - In two-dimensional (2-D) DNA typing, genomic DNA fragments are separated, first according to size by electrophoresis in a neutral polyacrylamide gel and second according to sequence by denaturing gradient gel electrophoresis, followed by hybridization analysis using micro- and minisatellite core probes. The 2-D DNA typing method generates a large amount of information on polymorphic loci per gel. Here we demonstrate the potential usefulness of 2-D DNA typing in an empirical linkage study on the red factor in cattle, and we show an example of the 2-D DNA typing analysis of a human pedigree. The power efficiency of 2-D DNA typing in general is compared with that of single-locus typing by simulation. The results indicate that, although 2-D DNA typing is very efficient in generating data on polymorphic loci, its power to detect linkage is lower than single-locus typing, because it is not obvious whether a spot represents the presence of one or two alleles. It is possible to compensate for this lower informativeness by increasing the sample size. Genome scanning by 2-D DNA typing has the potential to be more efficient than current genotyping methods in scoring polymorphic loci. Hence, it could become a method of choice in mapping genetic traits in humans and animals. PMID- 8250046 TI - Population genetics of the HRAS1 minisatellite locus. AB - Several years ago it was reported that rare HRAS1 VNTR alleles occurred more frequently in U.S. Caucasian cancer patients than in unaffected controls. Such an association, in theory, could be caused by undetected population heterogeneity. Also, in a study clearly relevant to this issue, it was recently reported that significant deviations from Hardy-Weinberg equilibrium exist at this locus in a sample of U.S. Caucasians. These considerations motivate our population genetic analysis of the HRAS1 locus. From published studies of the HRAS1 VNTR locus, which classified alleles into types, we found only small differences in the allele frequency distributions of samples from various European nations, although there were larger differences among ethnic groups (African American, Caucasian, and Oriental). In an analysis of variation of rare-allele frequencies among samples from four European nations, most of the variance was attributable to molecular methodology, and very samples from four European nations, most of the variance was attributable to molecular methodology, and very little of the variance was accounted for by nationality. In addition, we showed that mixture of European subpopulations should result in only minor deviations from expected genotype proportions in a Caucasian database and demonstrated that there was no significant deviation from Hardy-Weinberg equilibrium in our HRAS1 data. PMID- 8250047 TI - Extended multipoint identity-by-descent analysis of human quantitative traits: efficiency, power, and modeling considerations. AB - Goldgar introduced a novel marker-based method for partitioning the variation of a quantitative trait into specific chromosomal regions. Unlike traditional linkage mapping methods, Goldgar's method does not require the estimation of statistical quantities characterizing each locus thought to influence the trait under scrutiny (e.g., allele frequencies, penetrances, etc.). Goldgar's method is thus more flexible and less model dependent than many traditional marker-based genetic analysis techniques. Unfortunately, however, many of the properties of Goldgar's method have not been investigated. In this paper, the utility of an extended version of Goldgar's approach is studied in settings in which sibships are taken as the sampling unit of interest. The extensions discussed resolve around the incorporation of a wider variety of effects and factors into Goldgar's basic model. Analytic studies pertaining to power, sample-size requirements, and estimation procedures for the proposed extended version of Goldgar's method are described. Hypothesis-testing strategies are also discussed. The results of the analytic studies indicate that, although an extended sib-pair version of Goldgar's variance-partitioning approach to modeling the chromosomal determinants of a quantitative trait will be useful only for traits with high heritabilities or when fine-scale genetic maps can be employed. Goldgar's technique as a whole has promise, as it can be made relatively robust statistically, refined through some simple and intuitive extensions, and can be easily adapted to work with more complex sampling units. Further extensions of Goldgar's methods are proposed, and areas in need of additional research are discussed. PMID- 8250048 TI - Pedigree models for complex human traits involving the mitochondrial genome. AB - Recent biochemical and molecular-genetic discoveries concerning variations in human mtDNA have suggested a role for mtDNA mutations in a number of human traits and disorders. Although the importance of these discoveries cannot be emphasized enough, the complex natures of mitochondrial biogenesis, mutant mtDNA phenotype expression, and the maternal inheritance pattern exhibited by mtDNA transmission make it difficult to develop models that can be used routinely in pedigree analyses to quantify and test hypotheses about the role of mtDNA in the expression of a trait. In the present paper, we describe complexities inherent in mitochondrial biogenesis and genetic transmission and show how these complexities can be incorporated into appropriate mathematical models. We offer a variety of likelihood-based models which account for the complexities discussed. The derivation of our models is meant to stimulate the construction of statistical tests for putative mtDNA contribution to a trait. Results of simulation studies which make use of the proposed models are described. The results of the simulation studies suggest that, although pedigree models of mtDNA effects can be reliable, success in mapping chromosomal determinants of a trait does not preclude the possibility that mtDNA determinants exists for the trait as well. Shortcomings inherent in the proposed models are described in an effort to expose areas in need of additional research. PMID- 8250049 TI - Conclusions of segregation analysis for family data generated under two-locus models. AB - Susceptibility to a disease may involve the interactive effect of two genes. What conclusions will be drawn by segregation analysis in such a case? To answer this question, we considered a set of two-locus models and the corresponding exact distribution for 300 families. We investigated the conclusions and parameter estimations obtained for this sample, by comparing the likelihood expectations of the unified model and of more restricted models. In many cases, segregation analysis leads to the conclusion of a major gene effect, with or without a polygenic component--usually without a polygenic component in multiplicative models (i.e., where two genes have a multiplicative effect) and with such a component in nonmultiplicative models. For all the models considered, existence of a major gene effect is supported by transmission probability tests; there is evidence for transmission and agreement with the hypothesis of Mendelian transmission. Accordingly, there is no means of detecting that the effect of a major gene, with or without a polygenic component, does not correspond to the correct model. In addition, the parameter estimates for the major gene do not correspond to the characteristics of either of the two genes of the true model. This may substantially affect further linkage analysis. PMID- 8250050 TI - Our educational challenge. PMID- 8250051 TI - PCR-based cystic fibrosis (CF) carrier screening in a first-year medical student biochemistry laboratory. PMID- 8250052 TI - Knowledge of genetics among residents in obstetrics and gynecology. AB - A supervised genetics examination was administered to 76 obstetrics and gynecology (ob/gyn) residents from 15 different institutions in the Philadelphia area. The questions were specifically designed to be applicable to obstetrical practice. Overall, the mean score was 69% (range 32%-88%). Using the nonparametric Mann-Whitney rank sum test, we found that the 25 residents from institutions with an obstetrics-gynecology-genetics (OGG) program, coordinated by an obstetrician/gynecologist board certified in clinical genetics, had statistically significant higher scores than the remaining 51 residents from institutions without an OGG program (77% vs. 65%, respectively; P < .001). This study demonstrates that knowledge of genetics among residents in ob/gyn is deficient, especially among residents at institutions without OGG programs. Special efforts should be made to provide genetics education to these individuals. We propose that more obstetricians be encouraged to pursue postgraduate training in genetics in light of the rapid development of medical genetics; its application to prenatal screening, diagnosis, and counseling; the anticipated utilization of genetics services; and the need for educational initiatives geared toward ob/gyn patients. PMID- 8250053 TI - Involvement of rabbis in counseling and referral for genetic conditions: results of a survey. AB - Members of the New York Board of Rabbis were surveyed in the summer of 1991 to assess their activity in counseling congregants on issues related to genetics. Of a sample of 257 members, 181 (70.4%) responded to the questionnaire, and 175 of the responses were analyzed. More than half (56.0%) of the rabbis discussed health issues as a routine part of premarital counseling, and 22.3% had counseled a couple after prenatal diagnosis of an abnormal fetus. Orthodox rabbis were more likely than rabbis from other branches of Judaism to have contacted medical personnel in these cases, and they reported more involvement in helping families after the birth of a child with a hereditary condition or birth defect. However, a majority (90.9%) of rabbis from all branches would refer such a family for genetic counseling. Ninety-four rabbis (53.7%) discussed Tay-Sachs carrier testing with congregants. These rabbis tended to be Reform, to be younger, and to have fewer years in the rabbinate. Reform rabbis also scored significantly higher than did Orthodox or Conservative rabbis on knowledge questions about Jewish genetic diseases and were more active in distributing pertinent literature to congregants. Even though nearly 90% of the sample viewed counseling on genetic issues as part of their rabbinical role, most rabbis, even those who actually counseled on these issues, felt poorly prepared to do so. Recommendations are made for increased programming in rabbinical schools and for outreach from the genetics community. PMID- 8250054 TI - Educating the obstetric community through a genetic newsletter. PMID- 8250055 TI - On Historiography of Eugenics by Jonathan Marks. PMID- 8250056 TI - Chlorinated hydrocarbon solvents: substituting our way toward human carcinogenicity. AB - A discussion of the history of substitutions in industrial solvent use is presented, and animal carcinogenicity studies with certain chlorinated hydrocarbon solvents are reviewed. The present knowledge on the association between occupational exposure to chlorinated hydrocarbon solvents and human carcinogenicity is summarized, including a critical review of 23 studies pertaining to human carcinogenicity. Several deficiencies are identified in these studies that make interpretation of the results uncertain, including poor characterization of exposures in qualitative and quantitative terms, the small numbers of deaths or cases, and short follow-up periods. Despite these weaknesses, we conclude that, when viewed collectively, the evidence strongly suggests that occupational exposures to chlorinated hydrocarbon solvents are associated with an excess human cancer risk. The literature supports an association between occupational exposures to chlorinated hydrocarbons and elevated risks of leukemia, lymphoma, and urinary tract cancer. Parental occupational exposures are consistently associated with an elevated childhood cancer risk in the offspring. PMID- 8250057 TI - Organic dust exposures from compost handling: case presentation and respiratory exposure assessment. AB - Inhalation of dust from contaminated organic materials may result in acute respiratory tract illness. Possible mechanisms include toxic and cellular reactions to microbial and other organic products or immunologic responses after prior sensitization to an antigen. A case is presented of a 52 year old male who developed fever, myalgia, and marked dyspnea 12 hr after shoveling composted wood chips and leaves. Inspiratory crackles, hypoxemia, and bilateral patchy pulmonary infiltrates were seen. Precipitating antibody tests for the usual antigens were inconclusive. He improved over 3 days. In order to assess the environmental conditions the patient had experienced, we returned to the site to reproduce and measure respiratory exposures during hand loading of the compost. Visible clouds of fine particulate were easily generated during handling activities. Microscopic examination of these dusts indicated a predominance of spores. Endotoxin concentrations from inspirable and respirable dust samples ranged from 636 to 16,300 endotoxin units/m3. Levels of contaminants found were consistent with those associated with respiratory illness in other agricultural settings. Two respiratory disorders, hypersensitivity pneumonitis (HP) and organic dust toxic syndrome (ODTS), may occur after exposure to organic dusts containing fungal spores and endotoxins. Despite extensive clinical and environmental investigations, we were unable to differentiate these two disorders, and suggest they may represent parts of a spectrum of responses to complex organic dusts, rather than completely distinct clinical entities. PMID- 8250058 TI - Organic dust exposure from compost handling: response of an animal model. AB - The objective of this investigation was to elucidate the pulmonary responses of an animal model to dust generated from leaf/wood compost which had caused a severe case of acute respiratory illness in an individual. Guinea pigs were exposed for 4 hr to 30 mg/m3 of aerosolized leaf/wood compost dust. Inhalation resulted in significant cellular activation and changes in pulmonary mechanics. Maximal elevation in breathing rate (increases 36%) was observed 12-18 hr postexposure. Similarly, maximal granulocyte infiltration (increases 1,600%) and activation of alveolar macrophages (increases 65%) occurred 18 hr postexposure. In contrast, maximal airway obstruction (increases 120%) occurred immediately after exposure and returned toward normal (increases 53%) by 18 hr postexposure. In several respects, the airway obstruction and pulmonary inflammation described in the animal model were comparable to the human response to compost dust. Therefore, this animal model may be useful in predicting the potential respiratory hazard associated with exposure to various organic dusts. PMID- 8250059 TI - An environmental and respiratory health survey of workers in a grain mill in the Johannesburg area, South Africa. AB - A respiratory health survey was conducted in a grain mill and the relationship of health indicators to quantitative measures of airborne dust, fungal, and bacterial contamination was examined. Respiratory symptoms were more prevalent in the high dust exposure categories; lung function levels were also higher in the high dust exposure categories, consistent with a "healthy" worker effect. Workers in the three higher dust exposure categories showed either no change or a decrease in lung function over the working week, while workers in the low exposure category demonstrated an improvement in lung function over the working week. Total dust and microbiological (fungal and bacterial) load were found to be significantly related to each other, and the relationship of microbiological load to lung function level and changes over the working week were similar to those found for total dust. PMID- 8250060 TI - Relationship of respiratory health status to grain dust in a Witwatersrand grain mill: comparison of workers' exposure assessments with industrial hygiene survey findings. AB - Objective measures of exposure furnished by dust monitoring are both costly and time consuming and require a sufficient level of technology. However, they are important in demonstrating exposure-response relationships, in furnishing information necessary to establish environmental control levels, and to assess if interventions, for instance, improving dust control, have been effective. In this paper respiratory symptoms and cross-shift changes in spirometric lung function were related to dust exposure level in a grain mill assessed in two ways, subjectively (by workers themselves on a four point scale) and objectively (by personal dust monitoring). Health indicators that depend on the individual's perception (e.g., symptoms) correlated more closely with the subjectively assessed dust category, while health indicators that were measured objectively (e.g., cross-week FVC and FEV1 change) correlated more closely with the objectively assessed dust category. However, the patterns of relationship of respiratory health indicators to either dust category were similar, and exposure assessed by one method was, to a large extent, a proxy for the other. The most significant predictor of workers' choice of dust exposure category was the measured dust level. These findings indicate that exposure categories based on workers' assessment of dustiness can be a useful tool in etiologic research, in particular in establishing exposure-response relationships. PMID- 8250061 TI - Job tasks, potential exposures, and health risks of laborers employed in the construction industry. AB - Construction laborers have some of the highest death rates of any occupation in the United States. There has been very little systematic research focused exclusively on "laborers" as opposed to other workers in the construction industry. We reviewed the English language literature and various data bases describing the occupational tasks, exposures, and work-related health risks of construction laborers. The sources of information included 1) occupational mortality surveillance data collected by the states of California and Washington and the National Institute for Occupational Safety and Health (NIOSH); 2) National Occupational Exposure Survey; 3) national fatality data; 4) cancer registry data; and 5) case reports of specific causes of morbidity. While the literature reported that construction laborers have increased risk for mesothelioma, on-the-job trauma, acute lead poisoning, musculoskeletal injury, and dermatitis, the work relatedness of excess risks for all-cause mortality, cirrhosis, cerebrovascular disease, chronic obstructive pulmonary disease, ischemic heart disease, and leukemia is less clear. Furthermore, while laborers are known to be potentially exposed to asbestos, noise, and lead, and the NIOSH Job Exposure Matrix describes other potential hazardous exposures, little research has characterized other possible exposures and no research has been found that describes the exposures associated with specific job tasks. More advanced study designs are needed that include a better understanding of the job tasks and exposures to construction laborers, in order to evaluate specific exposure-disease relationships and to develop intervention programs aimed at reducing the rate of work-related diseases. PMID- 8250062 TI - Correlation between radiological and pathological diagnosis of silicosis: an autopsy population based study. AB - The radiological findings for the profusion of rounded opacities were compared to pathological findings for parenchymal silicosis in 557 gold miners who had, on average, 2.7 years between the radiological and pathological examination. Three readers read the radiographs, and ILO category 1/1 or more was defined as a positive diagnosis of silicosis. The sensitivity values were 0.393, 0.371, and 0.236, and the specificity values were 0.987, 0.965, and 0.978, for the three readers, respectively. The sensitivity of the readers improved with increasing degree of autopsy silicosis, but a large proportion of those with a moderate and marked degree of silicosis were not diagnosed radiologically. The diagnostic sensitivity of the radiological test could be improved by using category 0/1 as a cutoff point for workers exposed to a high average concentration of respirable silica dust. The diagnostic specificity of radiology could be improved by using category 1/0 or 1/1 as a cutoff point for a positive diagnosis for workers exposed to a low average concentration of respirable silica dust. PMID- 8250063 TI - Risk of silicosis in a cohort of white South African gold miners. AB - The risk of silicosis was investigated in a cohort of 2,235 white South African gold miners who had, on average, 24 years of net service from 1940 to the early 1970s and who were followed up to 1991 for radiological signs of onset of silicosis (ILO category 1/1 or more). There were 313 (14%) miners who developed signs of silicosis at an average age of 55.9 years. The latency period was largely independent of the cumulative dust exposure. In 57% of the silicosis, the radiological signs developed, on average, 7.4 years after mining exposure ceased. The risk of silicosis increased exponentially with the cumulative dust dose, the accelerated increase being after 7 mg/m3-years. At the highest exposure level of 15 mg/m3-years, which represents approximately 37 years of gold mining at an average respirable dust concentration of 0.4 mg/m3, the cumulative risk for silicosis reached 77%. In conclusion, the risk of silicosis was strongly dose dependent; however, the latency period was largely independent of the dose. PMID- 8250064 TI - A follow-up study of laboratory animal exposed workers: the influence of atopy for the development of occupational asthma. AB - In a 5-year follow-up study of 88 animal exposed laboratory technicians, the incidence of laboratory animal allergy (LAA), lung function, and the development of allergy test reactivity were investigated. Only two individuals developed test positive LAA rhinitis during the follow-up period. Furthermore, one subject who had previously had LAA rhinitis developed LAA asthma. In the remaining subjects the results of skin prick tests against laboratory animals and environmental allergens, total serum IgE levels, and lung function were unchanged. Atopy defined as parental and childhood allergy, raised total serum IgE levels, and positive skin prick tests against nonanimal environmental allergens and nonlaboratory animals (dog and horse) were risk indicators for development of test positive LAA asthma. The low incidence of LAA during the 5-year follow-up is interpreted as a result of an early LAA development in atopic subjects. PMID- 8250065 TI - Mortality patterns among men in the motor vehicle manufacturing industry. AB - This retrospective follow-up study evaluates the mortality experience of 123,232 male hourly workers employed at a motor vehicle manufacturing company in 1973. Company computerized personnel records were used to identify subjects and to obtain their employment history. Follow-up through December 31, 1985 was conducted using company, state, and national data sources. The mortality rates of the cohort were compared with the rates of the United States (U.S.) general population of white or black men, adjusting for age and calendar time, and using the standardized mortality ratio (SMR) as the measure of association. The overall mortality rate of cohort members is lower than the U.S. rate by 11% among whites (11,060 observed/12,427 expected deaths; SMR = 89) and by 24% among blacks (3,744/4,926 deaths; SMR = 76). These low mortality rates reflect deficits of diseases other than cancer. Cancer SMRs are 99 for whites and 90 for blacks. There are slight excesses of stomach cancer (SMR = 113) and of lung cancer (SMR = 108) among whites and of cancer of the large intestine and rectum (SMR = 114) and of the lymphopoietic tissue (SMR = 111) among blacks. Both whites and blacks have a large deficit of buccal cancer and pharynx cancer, and blacks also have a deficit of esophagus cancer. The cohort's overall deficit of deaths is due largely to the favorable mortality experience of active workers. In contrast, white subjects who have left active employment have an 11% mortality excess, and inactive black subjects have a mortality rate that is similar to the general population rate. The increased mortality among inactive whites is not limited to any one particular disease category. This pattern may reflect illness-related employment termination and generalized confounding by socioeconomic status, although the study does not rule out a relationship between workplace exposures and specific diseases. PMID- 8250066 TI - Retrospective follow-up study of foundry and engine plant workers. AB - A retrospective follow-up study of 21,013 workers employed at a foundry and two engine manufacturing plants was conducted to determine if these workers had an unusual mortality experience. A total of 2,235 deaths occurred during the follow up period of 1970-1987. Mortality from all causes was lower than expected. Men experienced a 6-13% excess of lung cancer deaths, depending on the choice of the comparison group. The data displayed evidence of a positive trend between lung cancer mortality and increasing duration of employment (p = 0.008). White men experienced a statistically significant excess of deaths from stomach cancer (standardized mortality ratio [SMR] = 158; 95% confidence interval [CI] = 101 234). Black men had increased mortality from pancreatic cancer, especially among engine plant workers (SMR = 303; CI = 121-624), and an excess of prostate cancer, concentrated among foundry workers (SMR = 234; CI = 112-430). PMID- 8250067 TI - Use of city directories as a source of occupational data in a case-control study. AB - The association between occupational factors and male breast cancer was investigated in a population-based case-control study conducted in Western New York state. Since 9% of cases and controls were missing occupational information in the tumor registry (usual occupation) and screening clinic (type of work done) files, supplemental occupational data were collected from commercial city directories. Occupational data were located for 69% of participants via the city directory thereby reducing the missing occupational data to 3%. For individuals with occupational listings in both the original source document and the city directories, similar titles were found for 94% of cases. Factors to be considered in assessing the feasibility of directory usage include purpose of study, degree of directory coverage within the geographic area, gender of participants, and availability of personnel and resources. City directories are a valuable supplemental source of occupational data for hypothesis generating case-control studies or registry-based studies of men conducted in urban areas. PMID- 8250068 TI - Clinical and histopathologic observations concerning hypotony after trabeculectomy with adjunctive mitomycin C. AB - Prolonged hypotony-induced maculopathy is a serious complication of trabeculectomy with adjunctive mitomycin C. We performed trabeculectomies with intraoperative mitomycin C on 59 eyes of 52 consecutive patients. Exposure time to mitomycin C was five minutes in the first seven patients, two of whom had prolonged hypotony-induced maculopathy. One of these required surgical revision of the filtering procedure. Light and electron microscopic study of the excised, avascular bleb disclosed an irregular epithelium and largely acellular subepithelium of loosely arranged connective tissue. In the remaining 52 eyes, the exposure time to mitomycin C was titrated between two and five minutes according to each patient's risk for failure of filtration from excessive fibrosis. Four additional cases of prolonged hypotony-induced maculopathy occurred among these 52 cases (7.7%), all of which were in the lower risk groups that received two- or three-minute exposure to mitomycin C. Four procedures failed, requiring further glaucoma surgery, and all of the patients were in the higher risk groups, receiving three- to five-minute exposures. Our titration of the exposure time to mitomycin C may have reduced, but did not eliminate, the risk fo prolonged hypotony-induced maculopathy, and further study is needed to establish the optimum protocol for the use of this drug as an adjunct to trabeculectomy. PMID- 8250069 TI - An evaluation of clusters in the glaucomatous visual field. AB - We used a statistical cluster analysis to analyze patterns of loss in 76 visual fields with typical glaucomatous defects to identify natural groupings of test locations in the visual field. Eleven clusters in the Octopus Program G1 (Interzeag, Inc., Northboro, Massachusetts) visual field were thus defined. In a separate population of 70 early glaucomatous and 70 age-matched normal visual fields, the local mean defects within these clusters and the global mean defect were calculated to assess their relative abilities to discriminate between the two groups. The 11 clustered mean defects collectively had a sensitivity of 90% and a specificity of 93%; the global mean defect had a sensitivity of 81% and a specificity of 91%. Additionally, we examined the long-term fluctuation clusters of test locations compared to the long-term fluctuation of individual test locations. Four hundred ten visual fields of 93 clinically stable eyes of 67 patients with glaucoma, as well as 210 visual fields of 105 eyes of 105 normal subjects were studied. In the stable glaucoma group, mean fluctuation of clustered test locations was 3.5 dB2, and mean fluctuation of individual test locations was 7.0 dB2. In the normal group, the respective values were 0.6 dB2 and 1.8 dB2. Cluster analysis was effective in detecting localized loss and in dampening long-term fluctuation. We studied the use of clusters in distinguishing normal from glaucomatous as well as stable from deteriorating visual fields. PMID- 8250070 TI - Corneal endothelial changes after excimer laser photorefractive keratectomy. AB - We used specular microscopy to investigate corneal endothelial changes after excimer laser photorefractive keratectomy. Twenty patients (26 eyes) underwent photorefractive keratectomy for the correction of myopia. The central cornea endothelium of each eye was observed preoperatively and one month postoperatively. Sixteen of the 26 eyes were examined by specular microscopy one year after photorefractive keratectomy. The corneal endothelial cell densities preoperatively, one month, and one year postoperatively were 3,221 +/- 216/mm2, 3,233 +/- 240/mm2, and 3,177 +/- 185/mm2 (mean +/- SD), respectively. The average coefficients of variation of mean cell area preoperatively and one month and one year postoperatively were 0.24 +/- 0.09, 0.23 +/- 0.05, and 0.22 +/- 0.05, respectively. There were no significant differences in mean cell density or the coefficients of variation of mean cell area among preoperative and one-month and one-year postoperative values. These results suggest that photorefractive keratectomy does not markedly affect the corneal endothelial cell density. PMID- 8250071 TI - The role of ciprofloxacin in endophthalmitis therapy. AB - Ciprofloxacin has been proposed for the systemic treatment of endophthalmitis. We studied the role of therapy based on the susceptibility of actual bacterial isolates and developed a system for assessing this susceptibility in endophthalmitis. Susceptibility testing was performed on 68 bacterial isolates from 66 patients with endophthalmitis. Our results indicated that, on the basis of a vitreous ciprofloxacin concentration of 0.25 microgram/ml, only 41 of 68 isolates (60%) would be inhibited by systemic therapy. All gram-negative bacteria (six of six) would be inhibited, whereas gram-positive bacterial susceptibility was variable. We concluded that systemic ciprofloxacin alone should not be administered empirically but it could provide an adjunct for treating selective cases of bacterial endophthalmitis. Bacterial susceptibility should be assessed on a blood serum standard of 0.25 microgram/ml instead of 1.0 microgram/ml. PMID- 8250072 TI - Comparison of topical antibiotics for treating Mycobacterium fortuitum keratitis in an animal model. AB - The efficacy of three topical antibiotic treatments for Mycobacterium fortuitum (strain ATCC-6841) keratitis were compared in rabbits. Rabbits were treated with ciprofloxacin (3 mg/ml) or clarithromycin (20 mg/ml) or a combination of amikacin (100 mg/ml) and vancomycin (50 mg/ml). All three treatments significantly reduced the number of organisms in treated eyes compared to untreated, control eyes (all P values < .001). No significant difference in treatment efficacy was found between the three treatment groups (all P values > or = .48), although ciprofloxacin (3 mg/ml) was more effective than clarithromycin (20 mg/ml) after excluding outliers (P = .01). All treatments stabilized or reduced the size of stromal infiltrates after four days of therapy, whereas infiltrates continued to enlarge in untreated eyes. These results suggest that topical clarithromycin, topical ciprofloxacin, and combined amikacin and vancomycin may all be clinically useful for treating M. fortuitum keratitis. Both clarithromycin and ciprofloxacin were better tolerated than combined amikacin and vancomycin. This study supports the further development of clarithromycin, a new macrolide antibiotic, as a topical drug for treatment of M. fortuitum keratitis. PMID- 8250073 TI - Risk of decreased visual acuity in full-thickness idiopathic macular holes. AB - We studied 169 eyes of 149 patients with a full-thickness idiopathic macular hole to ascertain the risk factors of enlargement of the hole or the surrounding retinal detachment and subsequent decreased visual acuity. Eyes were divided as follows: group A, 48 eyes with no posterior vitreous detachment and a hole 400 microns or smaller; group B, 77 eyes with no posterior vitreous detachment and a hole 400 microns or larger; group C, 13 eyes with posterior vitreous detachment and a hole 400 microns or smaller; group D, 31 eyes with posterior vitreous detachment and a hole 400 microns or larger. In group A, the macular hole enlarged in 37 eyes (77%); the retinal detachment enlarged in 35 eyes (73%); and visual acuity decreased by two or more Snellen lines in 28 eyes (58%). These percentages were significantly higher than those in group B (P = .01, P = .01, and P = .01, respectively), group C (P = .02, P = .01, and P = .01, respectively), and group D (P = .01, P = .05, and P = .01, respectively). Eyes without posterior vitreous detachment with a full-thickness idiopathic macular hole 400 microns or smaller may have a greater risk of an enlarged macular hole, surrounding retinal detachment, and subsequent visual loss. PMID- 8250074 TI - Risk factors for development of rhegmatogenous retinal detachment in patients with cytomegalovirus retinitis. AB - We studied 259 patients to determine the time-dependent risk and risk factors for the development of retinal detachment in patients with cytomegalovirus retinitis. The six-month and one-year retinal detachment rates (by eye) were 11% and 24%, respectively. Increasing retinal surface involvement outside of the posterior pole and the presence of retinitis activity were found to be the two covariates that best predicted retinal detachment. Eyes with peripheral involvement greater than 25% had a fivefold risk for detachment, compared to eyes with 10% involvement. If there was retinitis activity and more than 25% peripheral (external to major vascular arcades) involvement, the risk increased to 24-fold. The presence of a fellow eye with retinal detachment was not an independent risk factor. These observations should help in the design of a prophylactic trial intended to prevent retinal detachment and should also help ophthalmologists counsel patients with cytomegalovirus retinitis. PMID- 8250075 TI - Ophthalmic manifestations of infections with Cryptococcus neoformans in patients with the acquired immunodeficiency syndrome. AB - The present study was undertaken to determine the nature and the prevalence of ophthalmic manifestations of infections with Cryptococcus neoformans in human immunodeficiency virus seropositive patients and to analyze whether the presence or absence of ocular signs is associated with improved survival. Eighty human immunodeficiency virus seropositive patients with cryptococcal infection were enrolled. We observed papilledema in 26 of the 80 patients (32.5%). Visual loss and abducens nerve palsy occurred in seven patients (9%). Only two patients (2.5%) had optic atrophy. Visual loss caused by optic nerve involvement was less frequent among the 62 patients treated with oral conazoles exclusively than among the 18 patients who had received amphotericin B or a combination of amphotericin B and conazoles. Actual invasion of the intraocular structures with Cryptococcus neoformans was an uncommon complication in our series. In addition to the ocular manifestations attributable to cryptococcal disease, human immunodeficiency virus related retinopathy was present in nearly half of the patients. Cytomegalovirus retinitis was diagnosed in four patients (5%). The 26 patients (32.5%) with papilledema had a median survival of 182 days vs 160 days for the patients without papilledema. The median survival for 18 patients (22.5%) with cotton-wool spots was 102 days vs 186 days for those without cotton-wool spots. The differences between these subgroups were not statistically significant. PMID- 8250076 TI - Retinal lesions in septicemia. AB - We explored the association between septicemia and specific retinal lesions in a prospective controlled study. Hemorrhages, cotton-wool spots, or Roth's spots were found in 24 of 101 septicemic patients (24%), compared to four of 99 age- and gender-matched control patients (4%) (P = .0002). There was no significant association between types of organisms or focus of infection and the presence of specific lesions. Histologic examination of affected eyes disclosed cytoid bodies in the nerve fiber layer without inflammation. A definite association between septicemia and retinal lesions was found and indicates the need for routine ophthalmoscopy in septicemic patients. PMID- 8250077 TI - Relative letter and position difficulty on visual acuity charts from the Early Treatment Diabetic Retinopathy Study. AB - Ten Sloan letters were used in the visual acuity charts developed for use in the Early Treatment Diabetic Retinopathy Study. We used the data from the 3,710 Early Treatment Diabetic Retinopathy Study subjects to investigate the relative difficulty of the ten Sloan letters and to evaluate whether the position of a letter on a line affected its relative difficulty. In general, our findings were consistent with those of the previous study. The four letters with curved contours (C, O, S, and D) were more difficult to discern at threshold than the six letters (Z, N, H, V, R, and K) composed of straight lines. Our data demonstrate that under these test conditions, letters at the end of a line are more likely to be read incorrectly than letters at the beginning of the line. This finding indicates that these data are probably not useful for evaluating possible crowding phenomena. PMID- 8250078 TI - Comparison of Nidek 3Dx and Donaldson simultaneous stereoscopic disk photography. AB - We performed a retrospective study on 100 eyes of 55 consecutive patients to compare the stereoscopic quality of simultaneous stereoscopic disk photographs and the reproducibility of optic nerve assessments using the Nidek 3Dx system and the Donaldson fundus camera. Photographs were taken on the same eyes on the same dates and reviewed independently by one clinician three times in a randomized, masked fashion. Mean cup/disk ratios using the Nidek 3Dx system were similar to mean cup/disk ratios using the Donaldson camera. There was significantly less variability of disk analysis using the Nidek 3Dx system than the Donaldson camera (F = 1.6, P = .01, df = 99). Overall mean stereoscopic quality was significantly better with the Nidek system (3.2 +/- 1.15 S.D.) than with the Donaldson camera (2.5 +/- 1.27 S.D., P < .01). Age was the only variable that correlated with stereoscopic quality for both cameras. The new Nidek camera provided significantly better stereoscopic quality images than the Donaldson camera and with significantly more reproducible optic nerve head assessments. PMID- 8250079 TI - Growth of the human optic disk and nerve during gestation, childhood, and early adulthood. AB - We measured vertical and horizontal diameters of the optic disk and retrobulbar optic nerve in eyes from 95 patients on whom autopsies were performed at the UCLA Medical Center over a 20-year period. Ages at death ranged from 4.8 months' gestation to 21.9 years. Optic disk surface area and nerve cross-sectional area were calculated for each subject by using the formula for an ellipse. Approximately 50% of the growth of the optic disk and nerve occurs by 20 weeks' gestation, and 75% by birth; 95% of the growth occurs before the age of 1 year. All optic disk and nerve measurements correlate strongly (correlations > .67; P < .0001) with subject height and globe anteroposterior diameter. We applied our results to the current understanding of optic disk and nerve development, and compared them to previous clinical and pathologic studies of optic nerve dimensions in adults and older children. PMID- 8250080 TI - Hyperopia as a risk factor for nonarteritic anterior ischemic optic neuropathy. AB - The most convincing morphologic component in the pathogenesis of nonarteritic anterior ischemic optic neuropathy is the anatomically small and crowded optic nerve head. Because the appearance of the optic disk is presumed to be related to underlying refractive error, we sought to determine if a particular refractive error places patients at additional risk for, or affords protection from, nonarteritic anterior ischemic optic neuropathy. We compared refractive errors of 50 patients with nonarteritic anterior ischemic optic neuropathy to an age matched and eye-matched control population. The mean refractive error (in spherical equivalents) for the nonarteritic anterior ischemic optic neuropathy group was +0.26 diopter (SD +/- 2.08); the mean refractive error for controls was -0.86 diopter (SD +/- 2.91) (P = .027). Our results suggest that patients who have nonarteritic anterior ischemic optic neuropathy are less myopic than a control population. Whether it is myopia that protects from, or hyperopia that predisposes to nonarteritic anterior optic neuropathy, our results imply another anatomically based factor that characterizes eyes that develop nonarteritic anterior ischemic optic neuropathy. Those eyes tend to be minimally hyperopic. PMID- 8250081 TI - Optic disk risk factors for nonarteritic anterior ischemic optic neuropathy. PMID- 8250082 TI - Asymmetric pigmentary glaucoma caused by unilateral angle recession. PMID- 8250083 TI - Histopathologic characteristics of failed holmium laser sclerostomy. PMID- 8250084 TI - Indocyanine green angiographic findings in pseudoxanthoma elasticum. PMID- 8250085 TI - Anticardiolipin antibodies in isolated retinal vasculitis. PMID- 8250086 TI - Nadolol in the treatment of central serous retinopathy. PMID- 8250087 TI - Tuberculin-triggered panuveitis in a patient recently treated for active pulmonary tuberculosis. PMID- 8250088 TI - DNA diagnosis of Leber's hereditary optic neuropathy by using dried blood specimens. PMID- 8250089 TI - Lymphomatous optic neuropathy. PMID- 8250090 TI - Pseudotumor cerebri as a complication of amiodarone therapy. PMID- 8250091 TI - Marginal eyelid laceration after episiotomy. PMID- 8250092 TI - Calcium channel blockers in the management of low-tension and open-angle glaucoma. PMID- 8250093 TI - Genetic study of Japanese encephalitis viruses from Vietnam. AB - Two hundred eighty-five nucleotides from the capsid/premembrane gene region of 16 isolates of Japanese encephalitis (JE) virus from the southern and northern regions of Vietnam obtained during the period 1964-1988 were sequenced and compared with each other and with wild JE virus strains from China and Japan. The 16 Vietnamese isolates showed low rates of genetic evolution (< or = 3.2%) and were classified into one genotype with an overall similarity of > or = 95.4%, including the Japanese JaArOS982 strain taken as reference, and earlier isolated strains from Japan (Nakayama 1935) and China (Beijing-1 1949 and SA14 1954), the wild-type strains used to derive JE vaccines. The implications of the close genetic relationship among Vietnamese strains of JE virus analyzed in our study in relation to their epidemiology and evolution are discussed. PMID- 8250094 TI - The ubiquitin-ligase system in Trypanosoma brucei brucei. AB - The question of whether the African trypanosome Trypanosoma brucei brucei utilizes a ubiquitin-ligase system to conjugate the 8,500-dalton protein ubiquitin to other proteins has not been investigated. Using 125I-labeled ubiquitin and gel electrophoresis (sodium dodecyl sulfate and acetic acid, urea, Triton X-100), we looked for the incorporation of label into proteins larger than ubiquitin to determine ubiquitin-ligase system activity in cytosolic and nuclear lysates of long slender, intermediate, and short stumpy bloodstream-form trypanosomes. We present data suggesting that there is cytosolic activity of a ubiquitin-ligase system in all three bloodstream forms of T. brucei brucei. There are indications that the three bloodstream forms of T. brucei brucei differ in their cytosolic ubiquitin-ligase system activity. Our assay showed no activity of this system in the nucleus of T. brucei brucei. Further studies on the ubiquitin ligase system in T. brucei brucei may define differences between the three bloodstream forms, the parasite, and its host, leading to development of novel chemotherapeutic strategies. PMID- 8250095 TI - Immunochemical characterization and differentiation of two approximately 300-kD erythrocyte membrane-associated proteins of Plasmodium falciparum, PfEMP1 and PfEMP3. AB - Erythrocyte membrane-associated antigens of Plasmodium falciparum have been of long-standing interest as potential adherence receptors and vaccine candidates. We recently identified in trophozoite-stage infected erythrocytes a novel high molecular weight erythrocyte membrane-associated protein of P. falciparum, PfEMP3, defined by Western blotting with the rat monoclonal antibody 12C11. Genomic clone lambda 12.1.3 and cDNA clone p12.2 contain nucleic acid sequences encoding PfEMP3. Analysis of Malayan Camp strain parasites by sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 5% gels revealed that PfEMP3, defined by Western blot, has the same relative molecular weight (M(r)) as the surface-exposed protein PfEMP1 defined by cell surface iodination. We show here that PfEMP3 is distinct from PfEMP1 by three criteria. First, 125I-labeled PfEMP1 was resolved from PfEMP3 by extended migration on 4% gels. Second, in two strains of P. falciparum in which 125I-PfEMP1 has a different M(r), PfEMP3 had the same M(r). Third, immunization studies were performed with fusion proteins derived from clones lambda 12.1.3 and p12.2. Although one rabbit, Rb 05.75, immunized with the PfEMP3-derived fusion protein beta gal12.1.3, produced a serum that strongly immunoprecipitated PfEMP1 as well as PfEMP3, most sera immunoprecipitated only PfEMP3. Furthermore, immunoprecipitation of PfEMP3 by Rb 05.75 serum was blocked by the glutathione S-transferase 12.1.3 fusion protein, whereas immunoprecipitation of PfEMP1 was unaffected. Therefore, we conclude that PfEMP1 and PfEMP3 are antigenically distinct. PMID- 8250096 TI - Correlation between serum levels of antibodies to the 96-kD antigen of Plasmodium falciparum and protective immunity in Cameroon: a longitudinal study. AB - A longitudinal study was conducted in the Yaounde area of Cameroon that involved 211 individuals in June 1990, and 70 individuals for the follow-up study in December 1990. Sera from these subjects were tested against the recombinant 96 thermoresistant antigen of Plasmodium falciparum and the kinetics of antibody production to this protein show that titers tend to increase with age and are also related to antigen exposure. The increase in antibody titers with age correlates positively with the ability of the individual to prevent development of a high parasitemia. Adults who maintained stable high titers generally did not experience clinical attacks during the study period. The data suggest that antibodies against the 96-kD antigen participate in conferring some immunity to falciparum malaria. PMID- 8250097 TI - Immunoglobulin complex deposits in Plasmodium falciparum-infected placentas from Malawi and Papua New Guinea. AB - Term placentas from 35 patients infected with Plasmodium falciparum were obtained in Malawi in southeast Africa and six term placentas from patients infected with P. falciparum were obtained in Wewak, Papua New Guinea, Melanesia. The placental tissues were examined by light microscopy and by an immunohistologic method to compare the pathologic changes of placentas in the two malaria-endemic countries. Using the number of parasitized red blood cells (PRBC) in intervillous spaces, pregnant women from Malawi with placental parasitemia were categorized into three groups. In the high PRBC group (> 20%, group I), there was no deposition of IgE in fetal blood vessels. In contrast, IgE was observed in fetal blood vessels of the intermediate PRBC group (1-10%, group II) and low PRBC group (< 1%, group III). In all six placentas from Papua New Guinean women, deposition of immune complexes, including IgE, was observed in the fetal blood vessels. All placentas with deposition of IgE in fetal blood vessels showed no sequestration of malaria parasites in intervillous spaces. Our data indicate that the amount of deposition of IgE in the placenta from women infected with P. falciparum is inversely correlated with the degree of parasitemia at that site. PMID- 8250098 TI - Aggravation of both Trypanosoma cruzi and murine leukemia virus by concomitant infections. AB - Given the dissemination of acquired immunodeficiency syndrome (AIDS) in Latin America, where Chagas' disease is endemic, there is a present and increasing risk of concurrent infections with human immunodeficiency virus (HIV) and Trypanosoma cruzi. We used the model of murine acquired immunodeficiency syndrome (MAIDS) caused by a murine leukemia virus (MuLV) that induces immunologic alterations with similarities to those accompanying human HIV infection to study aspects of concomitant infections. The MuLV infection was found to reactivate T. cruzi infection in C57Bl/10 mice, as indicated by elevated parasitemia and lymphocytic infiltration in the myocardium. The T cells from these animals did not respond to T. cruzi antigens (lymphocyte proliferation, interferon-gamma, or interleukin-2 [IL-2] production) but had increased levels of IL-10. Trypanosoma cruzi-specific antibody was decreased but not absent in dually infected animals. In a second set of experiments, we infected MAIDS-resistant B6D2 mice with MuLV, followed by infection with T. cruzi. These animals had higher parasitemia than those infected with T. cruzi alone. More interestingly, only dually infected animals developed MAIDS. The present report describes the activation of T. cruzi infection by MuLV as well as the aggravation of MuLV infection by T. cruzi. These results may be relevant to coinfections with retrovirus and protozoan parasites in humans. PMID- 8250099 TI - Epidemiology of cutaneous leishmaniasis at a focus monitored by the multinational force and observers in the northeastern Sinai Desert of Egypt. AB - A longitudinal epidemiologic study of cutaneous leishmaniasis (CL) transmission was conducted between July 1989 and June 1991 in a 1,200-km2 sector of the northeastern Sinai Desert monitored by the Multinational Force and Observers (MFO), an international peace keeping mission between Egypt and Israel. The occurrence of human cases, sand fly density, rodent collection, and isolations of Leishmania confirmed only one of four surveyed locations as a significant focus of CL transmission. Phlebotomus papatasi, the only anthropophilic sand fly species encountered at this focus, comprised more than 96% of the sand fly population and attained human landing densities exceeding 100 sand flies/person/hr during 1990. Seasonal activity of this species ranged from April to November, with highest densities occurring during the period May-September. A peak promastigote infection rate of 2.4% (13 of 534) was observed in P. papatasi during July 1990. Twelve of the 60 (20%) persons at risk during the six months of intense sand fly activity at this site developed lesions consistent with CL; L. major was isolated from nine (75%) of these cases. Leishmania major infection was acquired by two of 22 (9%) sentinel hamsters used during the same period. More than 97% of the 897 wild rodents trapped at this site were desert gerbil species. Leishmania major was the only Leishmania isolated from human, sand fly, wild rodent (Gerbillus pyramidum), and sentinel hamster infections that originated at site Check point 1-Delta, the focus of CL transmission within jurisdiction of the MFO. The altered ecology of this area, created by construction of a dam, may contribute significantly to the transmission dynamics of CL at this focus. PMID- 8250100 TI - Application of seroepidemiology in identification of local foci in a malarious community in Palawan, The Philippines. AB - The value of seroepidemiology in malaria control programs has long been recognized, but its use in such situations has been limited. We therefore conducted a seroepidemiologic study in a frontier community on Palawan Island, The Philippines, an area where malaria is highly endemic. The first serologic survey was done at the end of the rainy season and the second was done during the dry season. The sera were examined using an indirect fluorescent antibody test. There was a significant difference in the geometric mean reciprocal titer (GMRT) during the rainy season as compared with the GMRT during the dry season. There were seasonal changes in the distribution of high- and low-titer responses, which is suggestive of the occurrence of recent past malaria epidemics. The differences in the geographic distribution of high and low titers were indicative of permanent foci of malaria transmission in this community. Results of a parallel parasitologic study were consistent with those of the serologic study. These findings may be of practical value not only in the proper reorientation of the local malaria control program, but of those of other endemic countries as well. PMID- 8250101 TI - Seroepidemiology of Coxiella burnetii among wildlife in Nova Scotia. AB - We used an indirect immunofluorescence assay to determine antibody titers to phase I and phase II Coxiella burnetii antigens in serum samples from a variety of wild animals in Nova Scotia. Forty-nine percent of the hares, 16.5% of the moose, 7.1% of the raccoons, and 1.5% of the white-tailed deer tested had antibodies to phase I antigen. We conclude that there is extensive infection of the hare population by C. burnetii, with lesser degrees of infection of the moose, raccoon, and deer population. PMID- 8250102 TI - Regression of diffuse intralobular liver fibrosis associated with visceral leishmaniasis. AB - Diffuse intralobular fibrosis of the liver is rare in cases of New World visceral leishmaniasis. A patient with this disease from a newly endemic area in the northernmost area of the Brazilian Amazon region was studied. Hypertrophy and hyperplasia of the phagocytic mononuclear cell system were observed, with parasitism and Disse's space fibrosis diffusely involving the liver. This description is based on a biopsy carried out after seven days of treatment with Glucantime. Another biopsy performed almost two years later showed no fibrosis, no Kupffer cell hypertrophy and hyperplasia, and no parasitism. The mechanism of fibrosis and regression is discussed. PMID- 8250103 TI - Use of Trypanosoma cruzi purified glycoprotein (GP57/51) or trypomastigote-shed antigens to assess cure for human Chagas' disease. AB - With the exception of assays for the detection of antibodies promoting complement mediated lysis of Trypanosoma cruzi, serologic tests have generally failed to assess the effectiveness of chemotherapy for Chagas' disease. Conventional serology, although useful for the diagnosis of infection, is not capable of determining which patients have been cured. Here we demonstrate that a high proportion of antibodies detected by conventional serology (using fixed epimastigotes or trypomastigotes or crude extracts obtained therefrom) are directed against the carbohydrate residue galactosyl alpha 1- > 3 galactose (Gal alpha 1- > 3 Gal), a determinant also recognized by antibodies from noninfected healthy volunteers. In a study of 14 cured patients with long-term followup, we found that the persistently positive reactions detected using conventional serology were largely eliminated following immunoadsorption with melibiose. Because of their wide distribution among microorganisms of intestinal and pulmonary microflora, these carbohydrate determinants may keep stimulating lymphocytes previously primed by T. cruzi Gal alpha 1- > 3 Gal epitopes, thereby accounting for false-positive results in cured patients. Consistent with this proposition, enzyme-linked immunosorbent assays performed with two distinct T. cruzi antigen preparations that lack the Gal alpha 1- > 3 Gal epitope, namely purified GP57/51 and trypomastigote-shed antigens, were indeed capable of determining a cure after chemotherapy, albeit to a different degree. Collectively, the data indicate that conventional immunoassays prepared with highly specific T. cruzi antigens can be useful in the assessment of a cure after chemotherapy. PMID- 8250104 TI - A polymerase chain reaction system for rapid diagnosis of scrub typhus within six hours. AB - A rapid diagnostic system for scrub typhus (tsutsugamushi disease) was established using the polymerase chain reaction (PCR) combined with microplate hybridization. With this system, diagnosis can be made within six hours without the need for radioisotopes, electrophoresis, or membrane hybridization. The use of a PCR processor and DNA-precoated enzyme-linked immunosorbent assay plates allows a rapid and precise diagnosis to be easily made even in primary medical care clinics. PMID- 8250105 TI - A randomized trial of ivermectin versus albendazole for the treatment of cutaneous larva migrans. AB - In an open study, we compared the efficacy of single doses of oral ivermectin (12 mg) and oral albendazole (400 mg) for the treatment of cutaneous larva migrans. Twenty-one patients were randomly assigned to receive ivermectin (n = 10) or albendazole (n = 11). All patients who received ivermectin responded and none relapsed (cure rate = 100%). All but one patient in the group receiving albendazole responded, but five relapsed after a mean of 11 days (cure rate = 46%; P = 0.017). No major adverse effects were observed. We conclude that a single 12-mg dose of ivermectin is more effective than a single 400-mg dose of albendazole for the treatment of cutaneous larva migrans. PMID- 8250106 TI - Amplification of quinine cardiac effects by the resistance-reversing agent prochlorperazine in falciparum malaria. AB - The use of reversing agents to overcome drug resistance is a potential new treatment strategy for both malaria and cancer. Laboratory studies have raised questions about the safety of this therapeutic approach, but data in humans are lacking. We therefore assessed the toxic potential of reversing agent therapy in Thai patients receiving quinine (17 mg/kg given over 4.5 hr) for falciparum malaria by serial measurements of the QTc interval, an electrocardiographic (ECG) marker of the effect of quinine. Six patients were randomly assigned to receive intravenous quinine alone while another six received one intramuscular injection of 12.5 mg of the reversing agent prochlorperazine (PC; compazine, stemetil) 2.5 hr after the quinine infusion had begun. Compared with baseline values at 2.5 hr, there was prolongation of the QTc interval 30, 60, 90, and 120 min after PC was injected (P < 0.05) but no further lengthening with quinine alone (P > 0.2). Prochlorperazine alone did not lengthen the QTc interval in six healthy volunteers. Neither total nor free quinine plasma levels increased after PC was injected, suggesting that ECG changes may have been due to PC-induced intracellular accumulation of quinine. Although only minor quinine ECG effects were amplified by the reversing agent PC in this study, resistance-reversing therapy could potentiate more serious drug effects. The possibility that more serious toxic effects could be produced by this therapeutic approach should be investigated further. PMID- 8250107 TI - Curing of chloroquine-resistant malaria with clindamycin. AB - A randomized comparative trial for treating adult patients with Plasmodium falciparum malaria was performed in Lambarene, Gabon. Forty-two patients received chloroquine (25 mg/kg for 48 hr) and 38 patients received clindamycin (5 mg/kg twice a day, for five days). Chloroquine treatment cured 15 patients (36%). Twenty patients (48%) showed recrudescent malaria by day 28 of follow-up (RI resistance) and seven patients (17%) showed persistent parasitemia after chloroquine treatment (RII/III resistance). In contrast, clindamycin treatment cured 37 of 38 patients (97%) and only one (3%) showed a recrudescence by day 28 (P < 0.001). Although the parasite clearance time was significantly longer after clindamycin treatment (median five days, range 3-6) than after chloroquine treatment (median four days, range 2-8) (P < 0.01), no differences were seen in the duration of symptoms after chemotherapy. In both treatment groups, no severe side effects occurred. Clindamycin can be used as a safe alternative to achieve radical cure in semi-immune adult patients with chloroquine-resistant P. falciparum malaria in Central Africa. PMID- 8250108 TI - [Reserves in decreasing maternal mortality due to obstetrical hemorrhages]. AB - Analysis of 542 lethal cases at pregnancy terms 28 weeks and more, of parturients and puerperae in Russia over 1987-1991 has shown that obstetrical bleedings were responsible for 42.2% of deaths, being the most frequent causes of lethal outcomes. Analysis of records documenting 283 lethal cases has shown a high incidence of somatic (90.9%) and obstetrical diseases (95.5%). Assessment of medica care quality has shown inadequate care of pregnant women and parturients and inadequate emergency care. Prevention of massive blood loss should involve hemostasis studies in pregnant women and parturients and its correction; if a bleeding occurs, comprehensive intensive care and surgical interventions are indicated. PMID- 8250109 TI - [The treatment of acute pyelonephritis in pregnant women]. AB - Analysis of 2058 pregnancy histories of patients with acute pyelonephritis helped work out and specify the indications for conservative and surgical treatment and characterize the etiopathogenetic factors and diagnostic methods. The necessity is emphasized of 'relieving' calix catheterization leading to elimination of retention shifts in the upper urinary tract and improving the efficacy of comprehensive antibacterial and detoxifying therapy. Methods and volumes of surgical interventions are substantiated. PMID- 8250110 TI - [The characteristics of the fetal-maternal-placental blood flow in habitual abortion]. AB - Dopplerometry of uterofetoplacental blood flow in 59 women at pregnancy terms of 24 to 40 weeks with a history of habitual abortions revealed disordered blood flow in approximately 60% of cases. Pathologic shifts first develop in a fetus as a rule. Moderate disorders of the uterofetoplacental blood flow in women with a history of habitual abortions do not result in the majority of cases in chronic disorders of fetal status, whereas marked disorders of blood flow often lead to fetal intrauterine suffering. PMID- 8250111 TI - [The hemostatic system in pregnant women and puerperae with the tetralogy of Fallot]. AB - The hemostasis system was studied in 31 pregnant women with Fallot's tetralogy, aged 18 to 32. In women with a history of radical correction of this condition a subcompensated form of the DIC syndrome was detected, manifesting as chronometric and structural hypercoagulation and platelet hyperaggregation. In patients with the cyanotic form of Fallot's tetralogy hypocoagulation thrombophilia was revealed, that was due to deficiency of contact factors (Hageman factor, highly molecular kininogene) and that was responsible for thrombohemorrhagic complications in these patients. Replacement therapy is recommended for such cases in labor and controlled anti-thrombotic therapy in the postpartum period. PMID- 8250112 TI - [The incidence of pathological microbial colonization and the adaptational characteristics of newborns with intrauterine hypotrophy]. AB - A total of 103 full-term small-for-date newborns were examined, as was a control group of normal full-term babies born to healthy mothers. Bacteriologic studies carried out in 148 newborns revealed most frequently Klebsiella, enterobacteria, Proteus, staphylococci, yeast fungi, and various associations thereof. If such microflora is found in small-for-date newborns, prophylactic antibiotic therapy is advisable to prevent the development of grave infections. PMID- 8250113 TI - [Controversial and unresolved problems of genital endometriosis]. PMID- 8250114 TI - [Renal hemodynamics studied by Doppler echography in newborn infants]. AB - The author describes a method for renal blood flow examination in the newborns making use of a lateral position of the sensor. Under study were linear blood stream velocity, time mean maximal velocity and time mean velocity, resistance index; renal artery diameter was measured and the minute and stroke renal perfusion was calculated. Three basic types of dopplerographic curves were distinguished: those characteristic of normal blood flow, of renal vascular spasm (no diastolic blood flow), and of drastic microcirculation disturbances (pendulum like blood flow). Renal artery dopplerography is an informative method useful to elucidate renal blood flow status. PMID- 8250115 TI - [The psychiatric aspect of the climacteric syndrome: its clinical picture and treatment]. AB - A total of 130 patients with the climacteric syndrome associated with mental disorders were examined by psychiatrists. Two groups of patients were distinguished. In Group 1 specific climacteric disorders ('flushes', excessive sweating, etc.) were the major components, whereas mental disorders (asthenic, phobic, hysterical, depressive) were just concomitant. In Group 2 the climacteric syndrome resulted in development of various mental diseases, mostly neuroses, depressions of various origins, superworship ideas. Recommendations on psychotropic therapy of these disorders were developed and contraindications against the use of hormonal therapy formulated. PMID- 8250116 TI - [Changes in the content of phospholipids and their metabolic products in the blood and tissues of patients with endometriosis interna and cancer of the corpus uteri]. AB - Levels of phospholipids, phosphoinosites, phosphatidylinosite-3-phosphates, diacylglycerols, arachidonic acid, prostaglandins E, F, J, and thromboxane A2 in patients with endometriosis and cancer of the corpus uteri were found to differ from those in healthy women. Biochemical parameters of whole blood and myometrial tissue of endometriosis patients differ significantly from those of patients with corpus uteri cancer. The authors suggest that reduction of phosphatidylinosite and phosphatidulinosite-3-phosphate levels in whole blood and their parallel increase in myometrial tissue are conducive to cellular proliferation and malignant degeneration. PMID- 8250117 TI - [The immunocorrective therapy of cervix uteri diseases]. AB - Analyzes the results of comprehensive clinical and immunologic examinations of women with inflammatory and tumorous diseases of the cervix uteri. Demonstrates a reduction of the immunologic reactions in such diseases and in traditional antibiotic therapy thereof. Presents methods of immunocorrective therapy with vitamin E, thymactin, and sodium nucleinate to be carried out on an inpatient basis. PMID- 8250118 TI - [The characteristics of hypophyseal gonadotropic function in patients with hypogonadotropic amenorrhea against a background of pergonal ovulation stimulation]. AB - Forty-three ovulation cycles stimulated with human menopausal gonadotropin were examined in 31 patients with hypogonadotropic amenorrhea. Peripheral blood estradiol and luteinizing hormone were radioimmunoassayed. The findings indicate the possibility of recovery of adenohypophyseal gonadotropin autosecretion in the presence of human menopausal gonadotropin administration. PMID- 8250119 TI - [Experience with using laser surgery in the treatment of benign diseases of the cervix uteri when it is deformed]. AB - Contact AYG laser was used in surgery on the deformed cervix uteri for cervicosis. The suggested method of using a contact laser scalpel at the main stages of the operation for partial stratification and miniconization provides a reliable hemostasis, minimal injury to the tissues, sterility of the operation site, and in stratification of the cervix uteri--a sufficiently easy dissection of the desired layer. The authors emphasize the usefulness of a differentiated approach to the choice of the method of surgical intervention (conization, plasty by partial or complete stratification), permitting the surgeon to combine the radical character of a surgical intervention with sparing of the anatomical shape of the cervical canal, and thus providing functionally stable results. PMID- 8250120 TI - [The use of tumor markers in the diagnosis and treatment monitoring of malignant tumors of the female genitalia]. AB - Levels of carcinoembryonic antigen (CEA), CA-19-9, CA-125, and mucin-like associated tumor antigen (MAA) were measured in the blood sera of 108 patients with uterine cervix and body, and ovarian carcinomas before treatment, and in those with ovarian carcinoma over the course of treatment to monitor its efficacy. The findings indicate that neither CEA nor MAA were diagnostically informative in malignant tumors of female genitals, and in uterine cervix cancer none of the tumor markers was diagnostically valuable. In neglected cases of uterine body cancer blood levels of CA-19-9 and CA-125 were elevated. In ovarian carcinoma CA-125 levels varied statistically significantly (p < 0.05) in various stages of the disease. CA-125 measurements are recommended for monitoring the treatment efficacy in patients with ovarian cancer. PMID- 8250121 TI - [THe indices of the immune status and cell membrane function in pregnant women with obesity]. PMID- 8250122 TI - [The characteristics of the microbiocenosis and local immunity of the intestines in newborns with an intrauterine infection]. PMID- 8250123 TI - [The clinical x-ray characteristics of breast status in patients with polycystic ovaries]. PMID- 8250124 TI - [Diagnostic and surgical hysteroscopy]. PMID- 8250125 TI - [The first obstetrical manuscript in the Russian language]. PMID- 8250126 TI - [The local anti-infection immunity of the female genital system]. PMID- 8250127 TI - [Tumor necrosis factor in the immunology of reproduction]. PMID- 8250128 TI - [The procedures of a neonatal service at a perinatal center]. AB - The authors analyze the development histories of 54 full-term newborns born in a satisfactory state, though in need of intensive care and rehabilitation at a special department for neonatal diseases. The absence of asphyxia at birth could not preclude a disease in the future. Analysis of mother's risk factors and neonatal disease permit a conclusion that babies born to mothers with a high-risk pregnancy even when born full-term should be singled out as a special group because of a high risk of disease development (intrauterine infection, cerebral disturbances, developmental defects, endocrine diseases, metabolic disorders, etc.). The most significant of mother's risk factors are reproductive function abnormalities (sterility, habitual abortions, a history of perinatal loss, etc.). Analysis of erroneous policy of neonatologists and late treatment start demonstrated that maternal risk factors should be more thoroughly assessed at childbirth. PMID- 8250129 TI - [The combined obstetrical and nephrological examination of pregnant women with kidney diseases and arterial hypertension]. PMID- 8250130 TI - [The use of plasmapheresis in the combined treatment of pregnant patients with pyelonephritis]. AB - The results of pyelonephritis treatment in 48 pregnant patients are analyzed. Twenty-three patients were administered routine antibacterial therapy, in 25 ones plasmapheresis was added to it. Immunity status of all the patients was examined. Antibiotic therapy was conducive to improvement of the patients' status, to a reduction of the renal inflammation severity, though it did not completely eliminate immunodeficiency signs. B lymphocyte and immunoglobulin levels persisted reduced. Plasmapheresis promoted a more swift elimination of clinical and laboratory signs of pyelonephritis and virtually completely liquidated immunodeficiency. PMID- 8250131 TI - [Intrauterine lavage in the treatment and prevention of puerperal endometritis]. AB - Addition of intrauterine lavage with cooled furacilin solution (0-4 degrees C) to a complex of treatment of 87 patients with postpartum endometritis resulted in a sooner recovery as against that in 61 patients administered traditional treatment alone; such a therapeutic course required lesser amounts of drugs, too. To specify endometritis severity and monitor the treatment efficacy, metro-aspirated specimen cytology was examined. Preventive lavage of the uterine cavity postpartum in cases with a 'dry' period longer than 24 h and other factors of risk of infectious complications proved to be highly effective. PMID- 8250132 TI - [The pathogenetic mechanisms of hyperthermia in patients with the premenstrual syndrome]. AB - Functional status of the central and autonomic nervous system was analyzed in 28 patients with the premenstrual syndrome one of whose manifestations was disordered thermoregulation. The examinations were carried out during both cycle phases. Analysis of the EEG data and clinical findings has shown different patterns of autonomic thermoregulation disturbances in three groups of patients and permitted the authors to suggest that changed activity of certain neurotransmitter systems connected with hormonal secretion over the course of the cycle was responsible for these disturbances. PMID- 8250133 TI - [New data on the pathogenesis of endometriosis interna]. AB - Eighty-six patients with histologically verified internal endometriosis were examined using histochemical, morphometric methods, and measurements of blood and myometrial tissue phosphatidyl inosites. Blood levels of phosphatidyl inosites were found reliably decreased in the patients vs. normal subjects, whereas their levels in myometrial tissue were significantly increased. Increased counts of tissue basophils, microcirculation disorders, lymphostasis and membranogenic stromal edema development were observed round endometriosis foci. Changes in the basic substance and fibrous structures of myometrial connective tissue complex were revealed, most marked in the areas adjacent to the endometrium. A possible mechanism of the development of pathologic shifts in internal endometriosis is discussed. PMID- 8250134 TI - [The morphofunctional characteristics of the amnion in normal and pathological pregnancies]. PMID- 8250135 TI - [The dynamics of the rapid changes in the phospholipids of the whole blood and blood cells in patients with endometriosis interna]. AB - Time course of rapid changes in blood phospholipids and blood cells was studied in 44 patients with endometriosis. Rapid reversible blood phospholipid changes were observed when these patients' red cells and lymphocytes were incubated with calcium chloride. Time course of rapid changes in phosphoinosites, sphyngomyelines, phosphatidylserines, phosphlatidylethanolamines of patients' red and white blood cells, whole blood, and myometrial tissue differed significantly from that in healthy women. The authors suppose that cellular proliferation processes in endometriosis are connected with phospholipid homeostasis disorders in blood cells and tissue. PMID- 8250136 TI - [The role of chlamydial infection in the genesis of tuboperitoneal sterility in women]. AB - The incidence of chlamydial infection in women with tuboperitoneal sterility is 49.5%. Clinically inflammatory diseases of the pelvic organs in such an infection are characterized by a primary chronic course with frequent recurrences and involvement of the cervix uteri, urethra, internal genitals, with moderately manifest clinical symptoms. Therapy of chlamydial infection should be combined with individual selection of therapeutic measures with due consideration for the inflammatory process phase and site, for the type of pathologic changes, general and immune status. PMID- 8250137 TI - [The treatment of bacterial vaginosis]. AB - A two-staged procedure involving creation of a physiologic vaginal environment and formation of normal microbiocenosis is suggested for correction of dysbiotic conditions of the vagina. Biologic preparations lactobacterin, acilact, bifidumbacterin, bifidin are used at the final stages of vaginitis therapy. Altogether 88.4% of patients were cured after the first eubiotic course, 6.8% after 2-3 courses. In 4.7% of patients the treatment was hardly effective. The results demonstrate a high efficacy of eubiotic therapy of bacterial vaginosis. PMID- 8250138 TI - [The characteristics of hormonal contraception in women with hyperandrogenism]. AB - One hundred and ten fertile women with such androgenism signs as seborrhea, acne, and hirsutism were examined to elucidate the contraceptive and therapeutic efficacy of hormonal contraceptive drugs anteovin, marvelon, and diane-35. The duration of pill administration for each women depended on her cycle duration. All the drugs had a 100% contraceptive efficacy and exerted a therapeutic effect on androgenism symptoms. PMID- 8250139 TI - [Reconstructive plastic operations in ptosis and prolapse of the genitalia. The choice of the technic and the prevention of complications]. AB - The principal causes of recurring prolapses are inadequate choice of surgical technique and unqualified performance of the operation proper, as well as the absence of expert evaluation of the performance capacity of this patient population. The authors suggest pathogenetically, anatomically, and functionally validated methods of reconstructive surgery permitting restoration of the position and function of the genitals and the adjacent organs and helping preserve the generative function of fertile women. Approaches to prevention of recurrences are outlined. PMID- 8250140 TI - [The principles of preoperative preparation in genital fistulae of an inflammatory origin]. AB - Body defense potential was examined in 156 patients with genital fistulas of inflammatory origin. Addition of myelopid, an immunomodulating agent, to the scheme of preoperative treatment was experimentally validated. Preoperative preparation was assessed from the time course of the patients' general status, blood parameters, liver and kidney functions, and immunity status. PMID- 8250141 TI - [The results of studying the erythrokinetics in puerperae suffering from iron deficiency anemia]. PMID- 8250142 TI - [The role of dynamic echography of the kidneys in an uncomplicated course of pregnancy]. PMID- 8250143 TI - [Endogenous opioid peptides in pregnancy and labor]. PMID- 8250144 TI - Evoked and spontaneous otoacoustic emissions. A comparison of neonates and adults. AB - Evoked otoacoustic emissions (EOAEs) and spontaneous otoacoustic emissions (SOAEs) were compared between neonates and adults. Several differences were seen for EOAEs: (i) the occurrence of EOAEs was significantly greater in adults than in neonates, and in neonates it increased with extra-uterine age; (ii) the high frequency EOAE intensity was greater in neonates than in adults; (iii) the EOAE intensity was significantly greater in neonates than in adults. No age-related differences for SOAEs were found: (i) age was not significantly related to the presence/absence of SOAEs in neonates or adults; (ii) the occurrence of SOAEs was, however, significantly greater in females than in males in both neonates and adults. PMID- 8250145 TI - Somatosensory evoked potentials in hemimegalencephaly and lissencephaly: anatomo functional correlations. AB - Four patients with hemimegalencephaly and four with lissencephaly were studied by median nerve somatosensory evoked potentials. The peripheral conduction velocity and the conduction time between the brachial plexus (Erb's point) and cervical cord were normal in both malformations. In hemimegalencephaly, the cortical components were absent on the malformed hemisphere but normal on the healthy side, whereas in lissencephaly an abnormal, prolonged N20 wave was recorded bilaterally. The difference presumably relates to the absence of normal cortical lamination in hemimegalencephaly and to the presence of a 'four-layered' cortex in lissencephaly. PMID- 8250146 TI - Phenobarbital in newborns with neonatal seizures. A study of plasma levels after intravenous administration. AB - Phenobarbital plasma levels were studied in a group of 25 newborn infants. Phenobarbital was administered i.v. in all cases throughout the study period. The mean loading dose was 19.4 mg/kg, ranging from 16.4 to 20.5, and the mean maintenance dose was 4.0 mg/kg/day, varying from 2.6 to 5.0. We obtained mean plasma levels of 22.9 micrograms/ml, 24 h after administering the loading dose. Mean plasma levels at 4, 7, 14 and 21 days were in the therapeutic range (15-40 micrograms/ml), with only a few cases falling outside of it. There was no difference in plasma phenobarbital levels between term and pre-term infants. Side effects were not seen in infants without a severe neurological impairment prior to drug administration. PMID- 8250147 TI - Maturation and effect of stimulus rate on brainstem auditory evoked potentials. AB - Brainstem auditory evoked potentials (BAEPs) at three stimulus rates (20, 41.3 and 61.3 c/s) were recorded in 104 normal preterm infants, distributed in four age groups between 32 and 39 weeks (conceptional age), and in 20 normal adults. The latency shifts between 61.3 c/s and 20 c/s, and between 41.3 c/s and 20 c/s, were calculated for each wave, and studied as a function of age. A significant effect of conceptional age was shown on the '61.3-20 c/s' and the '41.3-20 c/s' wave V latency shifts, and a significant difference between adults and newborns was noted for the wave V and wave III latency shifts. No significant difference was obtained for the wave I latency shifts. This suggests that the changes of auditory adaptation with maturation may be predominantly at a central level, i.e. on central synapses. PMID- 8250148 TI - Ependymal abnormalities in cerebro-hepato-renal disease of Zellweger. AB - The ependyma in six infants dying with cerebro-hepato-renal (CHR) disease showed similar but less extensive morphological and immunohistochemical abnormalities to those previously demonstrated in lissencephaly/pachygyria. More than two-thirds of the ependyma lining all ventricles was a pseudostratified columnar epithelium resembling midfetal life. Discontinuities did not correlate with minimal ventriculomegaly. Subventricular rosettes were common and not confined to regions of gaps in the overlying ependyma. Subependymal gliosis and glial nodules were absent. Immunoreactivity of ependymal cells for vimentin and GFAP was normal for age, but abnormally positive for S-100 protein and cytokeratin, as in lissencephaly; unlike lissencephaly, the rosettes in CHR disease also are reactive. Ependymal abnormalities may contribute to the pathogenesis of cerebral dysgenesis in CHR disease as in other genetic disorders of neuroblast migration. PMID- 8250149 TI - Prenatal diagnosis of GM2-gangliosidosis. Immunofluorescence analysis of ganglioside GM2 in cultured amniocytes by confocal laser scanning microscopy. AB - A confocal laser scanning microscopic system was used to detect the storage of ganglioside GM2 in Tay-Sachs fibroblasts and amniocytes. The diagnosis of the disease was confirmed by counting immunoreactive cells or by digital imaging analysis. This novel system facilitates the prenatal diagnosis of GM2 gangliosidosis. PMID- 8250150 TI - Sequential MRI findings in a patient with a germ cell tumor in the basal ganglia. AB - Serial changes of MRI scanning of an 11-year-old boy with hemiparesis due to a germ cell tumor in the basal ganglia are presented. Initial brain MRI T1-weighted images revealed a subtle mixed signal intensity lesion at left anterior and posterior limbs of the internal capsule. This lesion was not enhanced with Gd DTPA, however, T2-weighted images showed a misty high signal intensity lesion in the same region. The MR images also showed hemiatrophy of the left basal ganglia. Histologic examination of biopsy specimens of the tumor revealed an embryonal carcinoma. The patient was treated with chemotherapy with subsequent improvement in the hemiparesis. Our case suggests that germ cell tumors must be considered in the differential disorders of lesions in the ipsilateral hemiatrophic basal ganglia in pediatric patients with hemiparesis. PMID- 8250151 TI - Magnetic resonance imaging and positron emission tomography of band heterotopia. AB - A case of band heterotopia was reported with findings of positron emission tomography (PET). The patient was an 8-year-old girl who had mild mental retardation and intractable partial epilepsy. Her MRI showed another diffuse layer of gray matter underlying the normal-looking cortex and separated from it by an apparently normal layer of white matter. PET scan with [18F]fluorodeoxyglucose revealed that band heterotopia had the same degree of glucose metabolism as that of the overlying cortex. PMID- 8250152 TI - Mental retardation with marfanoid syndrome: presentation of a family with different phenotypical expression. AB - Here we report a new case in which the clinical manifestation were compatible with the phenotype described by Lujan et al. [Am J Med Genet 1984; 17: 311-22] as 'X-linked mental retardation with marfanoid habitus'. Based upon the presence of mild psychomotor retardation, epilepsy and skeletal malformations, a sister can be considered an affected carrier, whereas an older brother showed skeletal abnormalities and juvenile glaucoma. The mother had bilateral palpebral ptosis with minimal mitochondrial abnormalities at muscle biopsy. PMID- 8250153 TI - Early onset dystonia decreasing with development. Case report of two children with familial myoclonic dystonia. AB - Two related girls had the onset of unilateral leg dystonia in the neonatal period and at 13 months, respectively. The dystonic signs subsided with motor development and resolved completely in one of the girls by the age of 5 years. There was no response to L-dopa. From 2-3 years of age segmental myoclonus with a shoulder girdle distribution appeared. Family investigation results were compatible with autosomal dominant myoclonic dystonia responsive to alcohol. The onset and resolution of dystonia have not been described previously. This disorder is genetically separate from torsion dystonia. No linkage has been found to the dopamine beta-hydroxylase gene locus. Genetically determined disorders of neurotransmission may add to our knowledge of the normal development of motor control and thus merit further study. PMID- 8250154 TI - Some excitatory and inhibitory factors involved in the epileptic state. AB - Many factors are involved in the determination of the epileptic threshold and the precipitation of epileptic seizures. The balance between the underlying excitatory and inhibitory mechanisms are often imperfectly understood. However, some of these factors must be fully considered if the management of those with epilepsy is to be effective. This applies especially to children. Certain types of epilepsy are genetically determined, and others are age-related. Development of the brain plays a crucial role in the changing liability to epilepsy. If these processes are disturbed by various lesions and diseases, such as metabolic and toxic disorders, protective mechanisms may suffer. Kindling and mirror foci, the role of the cerebellum, and reflex epilepsy will be discussed. There are some seizure-inducing factors which are susceptible to change, and can be as important as anti-epileptic drugs in controlling seizures. The use and limitations of these measures, and of drugs, will be considered. PMID- 8250155 TI - Appearance of wave III of auditory brainstem response after removal of a cerebellar tumor. AB - Findings for auditory brainstem response (ABR) before and after surgical removal of a cerebellar tumor in a 10-year-old female are presented. ABR improved markedly, although the tumor showed no direct invasion to the brainstem. The cause of the ABR change and the origin of wave III are discussed. PMID- 8250156 TI - Long-term observation of the changing brainstem auditory evoked potentials in a case of infantile opsoclonus-polymyoclonia syndrome. AB - Brainstem auditory evoked potential (BAEP) studies were repeatedly carried out on a boy with infantile opsoclonus-polymyoclonia syndrome, from 15 months to 39 months of age, the following being revealed: the I-III interpeak latency (I-III IPL) was more or less in the upper normal range until 24 months and then remained at the same level throughout the rest of the observation period, not showing physiological shortening with maturation. It should be noted that the prolongation of I-III IPL continued in spite of the disappearance of the opsoclonus. PMID- 8250157 TI - Electrical status epilepticus in childhood: treatment with short cycles of high dosage benzodiazepine (preliminary note). AB - A treatment of electrical status epilepticus during childhood with relatively high doses of benzodiazepines (BZ) in short cycles (3 weeks) is proposed for subjects shown to be responsive to BZs (diazepam bolus test: DZP test). This treatment gave positive results in 7 out of 8 patients responsive to the DZP test (87%). PMID- 8250158 TI - An unusual case of Duchenne muscular dystrophy. AB - A 10-year-old boy with Duchenne muscular dystrophy (DMD), with quite unusual clinical data, is presented. He was unable to walk until age 6, walked only for 9 months and then became wheel-chair bound. No dystrophin was present on muscle biopsy sections and a large deletion was found in the dystrophin gene. The deletion encompassed the central high frequency deletion region of the gene. Early developmental milestones may be delayed in DMD, but patients usually start to walk around 2-3 years of age. A delay of the extent in this case is very unusual. PMID- 8250159 TI - Type C Niemann-Pick disease. Detection and quantification of cholesterol accumulating cells in bone marrow. PMID- 8250161 TI - Italian Society of Child Neuropsychiatry. PMID- 8250160 TI - On the problem of essential and secondary moyamoya and vascular dysplasia in Down syndrome. PMID- 8250162 TI - Annual meeting of the European Academy of Allergology and Clinical Immunology 1993. Rotterdam, The Netherlands, 12-15 September 1993. Abstracts. PMID- 8250163 TI - Combined continuous spinal-epidural anaesthesia with a single interspace, double catheter technique. AB - A double catheter, single interspace combined spinal-epidural anaesthetic was performed in 12 ASA grade 3 patients. The technique failed in one patient in whom the intrathecal catheter could not be inserted satisfactorily. In another patient, the epidural catheter was accidentally placed intrathecally. The maximum height of the block obtained with hyperbaric lignocaine or bupivacaine was below T10 in all patients and there were no adverse cardiovascular changes. As opposed to a classical combined spinal-epidural block, this technique is most suitable for very debilitated patients. The advantages are the ability to titrate the intrathecal dose of the local anaesthetic to achieve the desired dermatomal level and to test the correct position of the epidural catheter before injecting drugs intrathecally. PMID- 8250164 TI - The levering laryngoscope. PMID- 8250165 TI - Anaesthesia for the porphyric patient. PMID- 8250166 TI - Anaesthesia for the porphyric patient. PMID- 8250167 TI - Pathogenesis of postimmersion collapse. PMID- 8250168 TI - Lipaemia affecting automated clotting results. PMID- 8250169 TI - Entonox-isoflurane for analgesia in labour. PMID- 8250170 TI - Laryngeal mask insertion--a useful tip. PMID- 8250171 TI - A tie for the laryngeal mask airway. PMID- 8250172 TI - Increased propofol requirements in a child with Duchenne muscular dystrophy. PMID- 8250173 TI - A problem with the Graseby 3300 PCA pump. PMID- 8250174 TI - Intravenous magnesium therapy in critically ill patients. PMID- 8250175 TI - A faulty epidural catheter. PMID- 8250176 TI - Breathing systems filters can affect the performance of anaesthetic monitors. PMID- 8250177 TI - Conversion of orotracheal to nasotracheal intubation with the aid of the fibreoptic laryngoscope. PMID- 8250178 TI - Pressure effects of syringes. PMID- 8250179 TI - Pressure effects of syringes. PMID- 8250180 TI - Bronchospasm during epidural anaesthesia. PMID- 8250181 TI - Propofol and electroconvulsive therapy. PMID- 8250182 TI - Horner's syndrome due to epidural anaesthesia presenting with a painful eye. PMID- 8250183 TI - Intra-operative heat loss--wrapping up the problem with a space blanket. PMID- 8250184 TI - 'Smelly agents'. PMID- 8250185 TI - Minimally invasive therapy--implications for anaesthesia. PMID- 8250186 TI - Neuromuscular effects and intubating conditions following mivacurium: a comparison with suxamethonium. AB - Mivacurium chloride has been assessed in respect of intubating conditions and neuromuscular effects. The influence of suxamethonium on the onset and duration of subsequently administered mivacurium was also studied. A dose of 0.15 mg.kg-1 of mivacurium was found to provide unacceptable intubating conditions at 2 min in 9/9 patients and further studies were conducted using 0.2 mg.kg-1. Intubating conditions with this dose were acceptable in 65% and 80% of patients at 2 min and 2.5 min respectively. In comparison, intubating conditions were acceptable in 100% of patients at 1 min following 1 mg.kg-1 of suxamethonium. The onset of block occurred in 96 s and 97 s after 0.15 mg.kg-1 and 0.2 mg.kg-1 respectively, and the durations of block in terms of recovery of the first twitch (T1) to 25% and 90% of control, and to recovery of train-of-four ratio to 0.7, were 16.1 and 17.9; 24.1 and 25.8; and 24.2 and 27.0 min respectively with the two doses. The time for the onset of complete block with suxamethonium 1.0 mg.kg-1 was 50 s and the times to 25% and 90% recovery were 9.8 min and 13.3 min. The differences between suxamethonium and both doses of mivacurium were significant (p < 0.05) but there were no significant differences between the two doses of mivacurium in any of the neuromuscular measurements. Prior administration of suxamethonium had no influence on the effects of mivacurium. Cutaneous flushing was observed in 30 out of 75 patients but this was associated with transient hypotension in only two patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250187 TI - Personality traits as predictors of anxiety prior to caesarean section under regional anaesthesia. AB - One hundred and fifteen patients scheduled for elective Caesarean section under regional anaesthesia were questioned to determine their personality profiles and anxiety state. Positive correlations were found between neuroticism and anxiety scores (p < 0.05). Patients with tendencies towards both neuroticism and introversion had higher pre-operative anxiety scores than other personality types (p = 0.015). In general, previous experience did not reduce pre-operative anxiety and personality appeared to be a better predictor of anxiety prior to Caesarean section under regional anaesthesia. Some personality types might benefit from additional support to improve their ability to cope with this stressful event. PMID- 8250188 TI - Eltanolone for induction of anaesthesia and to supplement nitrous oxide for minor gynaecological surgery. AB - We have used eltanolone to induce anaesthesia and by intermittent injection to supplement nitrous oxide during maintenance in 50 patients undergoing minor gynaecological surgery; opioids were not used. Induction of anaesthesia was remarkably trouble free and achieved with a single dose of 0.5 mg.kg-1 in 90% of patients. Pain on injection was not a feature. Satisfactory anaesthesia was achieved in 84% of patients; eight required addition of isoflurane. There were minimal cardiovascular changes and apnoea on induction occurred in only one patient. Recovery times were acceptable. PMID- 8250189 TI - A comparison of the effect of isoflurane and propofol on arterial carbon dioxide tensions following intracranial surgery. AB - Thirty-four adult patients undergoing elective craniotomy were randomly allocated to receive either isoflurane or propofol to supplement nitrous oxide and fentanyl for maintenance of anaesthesia. Arterial blood gases were measured pre operatively before premedication and at 5, 15, 30 and 60 min after tracheal extubation. The arterial carbon dioxide tension was higher than the preoperative value 5 min after tracheal extubation in both groups (p < 0.01), but there was no difference between the two groups at any time. It is concluded that the degree of postoperative respiratory depression after intracranial surgery is short-lived and minimal following maintenance of anaesthesia with either propofol or isoflurane. PMID- 8250190 TI - Plasma concentrations of lignocaine after thoracic paravertebral blockade in infants and children. AB - Plasma concentrations of lignocaine were determined in six ASA 1 children treated with thoracic paravertebral blockade. Peak concentrations ranged between 1.7 and 3.0 micrograms.ml-1 after a bolus injection of lignocaine 0.5 ml.kg-1 (lignocaine 10 mg.ml-1 with adrenaline 5 micrograms.ml-1) and peak plasma concentrations were reached 15-30 min after injection. Two hours after the initial bolus a continuous infusion of the same lignocaine solution was started (0.25 ml.kg-1.h-1). Steady state levels of lignocaine were reached 8-10 h after the start of the infusion and ranged from 2.1-3.2 micrograms.ml-1. This regimen for paravertebral block in children resulted in moderate plasma concentrations of lignocaine, well below those which might result in side effects or systemic toxicity. PMID- 8250191 TI - Dexamethasone reduces pain and swelling following extraction of third molar teeth. AB - Extraction of multiple third molar teeth can cause significant postoperative pain, swelling and trismus, which may result in delayed hospital discharge. We have examined the effect of a single prophylactic dose of oral dexamethasone 8 mg on these complications, in a randomised double-blind study of 50 adult patients. The number of extractions performed and the operative approach were standardised. Dexamethasone resulted in a significant reduction in pain 4 h postoperatively, and eliminated the need for opioid analgesia in the postoperative period. The incidence of severe swelling was also reduced significantly, but there was no effect on trismus. Postoperative nausea and vomiting were significantly lower in the dexamethasone group. We conclude that the use of prophylactic oral dexamethasone is useful in reducing postoperative analgesia requirements in this group of patients, and may facilitate surgery performed on a day case basis. PMID- 8250192 TI - An ethnic comparison of the sympathetic response to tracheal intubation. AB - The sympathetic response to orotracheal intubation was examined in five Europeans, 15 Chinese, and seven Nepalese male patients. Plasma adrenaline and noradrenaline concentrations were assayed using high performance liquid chromatography with colorimetric detection. There was a significant rise in noradrenaline concentrations following intubation in the Chinese and the concentrations decreased over 5 min. Similar increases were seen in the Europeans and Nepalese. The mean plasma catecholamine concentrations were comparable, which suggests that there are no ethnic differences in the groups studied. A wide interindividual variation of catecholamine concentrations was found. PMID- 8250193 TI - Avoidance of intermittent positive pressure ventilation in tetanus with dantrolene therapy. AB - Two patients with features of severe tetanus who were successfully treated with dantrolene are reported. Neither patient required treatment with neuromuscular blocking agents so mechanical ventilation, with all its disadvantages, was avoided. PMID- 8250194 TI - Anaesthesia for cardiomyoplasty. AB - Cardiomyoplasty is a new surgical procedure for the treatment of severe heart failure, but early mortality has been high in the preliminary experience of many centres. This report describes the anaesthetic techniques used for the first two successful cases to be performed in the UK. PMID- 8250195 TI - Postoperative bilateral diaphragmatic paralysis and subsequent anaesthesia. AB - A case is described of a 56-year-old man who developed bilateral diaphragmatic paralysis following surgery remote from the course of the phrenic nerves. This rare complication is reviewed and the management of subsequent anaesthesia described. PMID- 8250196 TI - Subcutaneous emphysema, pneumomediastinum and pneumothorax complicating laparoscopic vagotomy. Report of two cases. AB - Two patients developed subcutaneous emphysema and pneumomediastinum during laparoscopic vagotomy. One of the patients also had a pneumothorax which produced a sudden increase in end-tidal carbon dioxide concentration preceding arterial oxygen desaturation. The pneumothorax was drained with an intercostal cannula. The patient required a twofold increase in minute ventilation to maintain normocarbia, probably because of the additional absorption of carbon dioxide through the pleural cavity. Despite the presence of a peritoneo-pleural communication, surgery was successfully completed. We believe that gas under tension in the peritoneal cavity dissected along tissue planes around the oesophagus opened up during surgery. Thus pneumomediastinum, subcutaneous emphysema and pneumothorax are definite risks associated with this new procedure. PMID- 8250197 TI - The intraosseous route for induction of anaesthesia. PMID- 8250198 TI - The Bicore pulmonary monitor. A device to assess the work of breathing while weaning from mechanical ventilation. AB - The Bicore CP-100 pulmonary monitor offers the advantage of continuous pulmonary monitoring at the bedside. Using an easily placed oesophageal balloon catheter and a flow transducer within the ventilator system, intrathoracic pressures, airway pressures and airway flow can be measured, and from these the Bicore calculates 25 other respiratory variables. The monitor screen displays real time waveforms of airway pressure, airway flow, oesophageal pressure and inspired tidal volume, and with these data it should be possible to determine more accurately when patients can be weaned from mechanical ventilation. It should also be possible to assess objectively the onset of respiratory fatigue and failure so that intervention can take place at an earlier stage. To assess the ease of use and potential benefit, the Bicore was used to calculate the work of breathing while weaning eight patients from mechanical ventilation in a spontaneous breathing mode. The pressure support ranged from 20 cmH2O, 10 cmH2O and 5 cmH2O, to a continuous positive airway pressure of 5 cmH2O, all with a positive end-expiratory pressure of 5 cmH2O, down to a single T-circuit with no positive airway pressure or end-expiratory pressure. The work of breathing while on a Veola Hamilton ventilator was found to be about the same, while receiving pressure support of 20 cmH2O compared to a continuous positive airway pressure of 5; the other forms of respiratory support increased the work of breathing significantly. PMID- 8250199 TI - Anaesthesia and rheumatoid arthritis. AB - A review of the implications of rheumatoid arthritis on peri-operative anaesthetic management is presented. Pre-operative assessment should include a careful search for articular and systemic manifestations of the disease that may complicate intraoperative care. Drug disposition may vary from the normal patient as a result of changes in serum protein binding, together with possible exaggeration of pharmacodynamic responses. Airway management presents one of the greatest challenges to the anaesthetist, and care in manipulation of the cervical spine is paramount. PMID- 8250200 TI - Blood ordering practices in obstetric units in the United Kingdom. AB - A postal questionnaire on blood ordering practices and blood availability was sent to 89 randomly selected maternity units within the United Kingdom. The replies demonstrated a wide variation in crossmatching practices. Of those units that replied, 56% crossmatch for elective and 64% for emergency Caesarean section, 54% for manual removal of placenta, and 29% for fetal distress in labour. The remainder 'group and screen' for these indications. Lack of 24 h cover by a resident haematology technician, and location of blood banks distant to the obstetric unit were associated with significant delays in blood availability. More extensive use of the 'group and screen' technique may reduce unnecessary crossmatching without jeopardizing patient safety, and is advocated for maternity units. PMID- 8250201 TI - [Diagnosis of malignant hyperthermia susceptibility. 1. The significance of in vitro susceptibility tests]. AB - Molecular genetic findings indicate genetic heterogeneity in susceptibility to malignant hyperthermia (MH). At present the in vitro contracture test (IVCT) is still the most reliable diagnostic procedure for MH susceptibility. It must be performed in a standardized fashion. METHODS. We investigated 350 patients (233 children and 117 adults) using the protocol of the European MH Group for the IVCT. The test results were classified as susceptible to MH (MHS), non susceptible to MH (MHS), non-susceptible to MH (MHN) and equivocal (MHE), with an abnormal caffeine result designated MHEc and an abnormal halothane result designated MHEh. Reasons for the IVCT were a positive family history for MH susceptibility (n = 94), a MH reaction (n = 157), creatine kinase elevation unknown aetiology (n = 53) and different neuromuscular diseases (NMD, n = 46). Physical, neurological and laboratory work-up included serum enzymes, nerve conduction studies, electromyography and muscle biopsy evaluated by different techniques. Thirty-one children and 11 adults were MHS, while 152 children and 80 adults were MHN. MHE findings were obtained in 50 children and 26 adults. While the MHS and MHN groups are diagnostically safe, the equivocal group is not, with possible false-negative or false-positive interpretation. The high number of MHE findings most probably is explained by the high proportion of patients with NMD (53% of the children, 69% of the adults). RESULTS. In a group of 18 boys with Duchenne or Becker muscular dystrophy, ranging in age from 1.5 to 24 years, the IVCT results were twice MHS, once MHE, and MHN in the remaining 15 cases. In seven other boys with Duchenne or Becker muscular dystrophy, proven by molecular techniques, there were anaesthetic complications with MH-like symptoms. After administration of trigger substances, five out of the seven suffered a cardiac arrest, two of whom died. In the surviving five boys the IVCT results were three times MHN, once MHE and once MHS. Most probably these boys suffered from effects of succinylcholine, possibly potentiated by other trigger substances. The adverse cardiac reactions are attributed to triggered rhabdomyolysis with associated hyperkalemia but not a primary hereditary disposition to MH. CONCLUSION: In patients with NMD, MHS and MHE test results do not indicate a hereditary, heterogeneous disposition to MH; the majority will be caused by a secondary induced disturbance of calcium homoeostasis in the diseased muscle cells. These results do, however, indicate the following: (1) Patients with NMD exposed to trigger substances are at higher risk than the general population for MH-like episodes, including sudden death. (2) NMD therefore should be diagnosed as early as possible and patients should not be exposed to trigger substances when alternatives are at hand. (3) Diagnostic procedures in patients having suffered an MH-like episode should include IVCT and special investigations to exclude or substantiate other NMD. The work-up may be changed if a family member is properly classified as MH susceptible. (4) In patients with known NMD there is no indication for performing IVCT, since the results may even be misleading. PMID- 8250202 TI - [Diagnosis of malignant hyperthermia susceptibility. 2. Anesthesia for muscle biopsy. Differential diagnosis in negative test results]. AB - At present the in vitro caffeine-halothane contracture test is the only test to predict susceptibility to malignant hyperthermia (MH) with acceptable sensitivity and specificity. Anaesthesia is necessary for the excision of muscle bundles from the vastus lateralis muscle. MATERIAL AND METHODS. Between 1983 and 1991, muscle biopsies were taken from 350 patients, 233 children and 117 adults. In 197 cases, trigger-free general anaesthetics were supplemented by ventilation, via mask in 124 cases and via tracheal intubation in 73 cases. One hundred and fifty-three muscle biopsies were performed under regional or local anaesthesia with or without sedation. Fourteen of the patients with regional anaesthesia needed an additional general anaesthetic to tolerate the operation. For premedication midazolam was administered orally. The general anaesthetics consisted of etomidate or propofol with fentanyl or alfentanil, always with nitrous oxide and oxygen. For local anaesthesia we used procaine or prilocaine. During the first 3 years local infiltration of the thigh was used, but subsequently direct nerve blockade of the femoral nerve and the lateral cutaneous femoral nerve was preferred. RESULTS. In all cases of general anaesthesia and in 91.6% of cases of regional anaesthesia operating conditions were very good. Fourteen (8.4%) of the patients with regional anaesthesia needed an additional general anaesthetic. There were no severe complications noticed. Moderate complications were found in 29 of the 194 paediatric general anaesthesias (15%) and in 5 of the 117 regional anaesthesias in adults (4%); all were easy to treat. The 50 paediatric regional anaesthesias and the three general anaesthesias in adults were without complications. DISCUSSION. For muscle biopsies, trigger-free general anaesthesia can be recommended as well as peripheral nerve blockades. Complete monitoring is necessary, even for this minor procedure: ECG, blood pressure, pulse oximetry, capnometry, measurement of body temperature and blood gas status. Differential diagnosis in negative test results. In patients who suffer an anaesthetic incident, the following disease must be considered: myopathies (especially the congenital myopathies and muscular dystrophies), respiratory problems due to pulmonary infection and obstruction, metabolic disorders of various origins, and the problem of masseter spasm. PMID- 8250203 TI - [The behavior of arterial and mixed venous oxygen and carbon dioxide partial pressure and the pH value during and following intubation apnoea. Studies on the occurrence of the Christiansen-Douglas-Haldane effect]. AB - The Christiansen-Douglas-Haldane effect describes the reduced CO2 binding capacity of oxygenated compared to deoxygenated haemoglobin. Under the condition of a "closed system", for example hyperoxic apnoea after adequate preoxygenation (continuous O2 uptake with lack of CO2 delivery), specific effects on the arterial and mixed venous blood gas status, due to the Haldane effect, are seen: within 30 s after onset of apnoea, "paradoxical pCO2" (paCO2 exceeds pvCO2) and "pH reversal" (pHa falls under pHv) can be observed. It was the aim of this study to demonstrate how fast arterial and mixed venous pCO2 and pH normalize when a change from apnoea ("closed system") to controlled ventilation ("open system") takes place. METHODS. 12 patients (ASA II-IV, NYHA II-III) scheduled for coronary artery bypass grafting were studied. Premedication consisted of flunitrazepam 2.0 mg p.o. given the evening before operation and another 2.0 mg p.o. given 90-120 min before induction of anaesthesia. Routine preparation for induction consisted of venous and arterial cannulas, pulmonary artery catheter and continuous pulse oximetry. Following standardized preoxygenation, induction of anaesthesia was performed with fentanyl, pancuronium and etomidate. After cessation of spontaneous respiration, controlled ventilation was continued with 100% O2 until intubation. Intubation and insertion of stomach tube and oesophageal temperature probe were undertaken after exactly 2 min. After reconnection to the semi-closed circle breathing system, controlled ventilation was continued with 100% O2. Eighteen arterial (a) and 18 mixed-venous (v) blood samples were drawn simultaneously in a sequential manner immediately before and during the last 20 s of apnoea, as well as within 4 min after onset of controlled ventilation (Table 1). The pO2 (mmHg), pCO2 (mmHg) and pH were determined using a Stat Profile 5 blood gas analyser. RESULTS. During apnoea and within the first 35 s of controlled ventilation the paO2 showed a total decrease of 131.5 mmHg that was followed by an almost linear increase of 29.7 mmHg/min (Fig. 1a). In the course of apnoea and controlled ventilation the pvO2 remained relatively stable, with values ranging from 42 to 43 mmHg (Fig. 1b). During apnoea the paCO2 showed an increase of 12.5 mmHg that was followed by a biphasic decrease (first 13.8 mmHg/min and then 0.75 mmHg/min) beginning 15 s after the onset of controlled ventilation (Fig. 2a). With an increase of 4.2 mmHg, the pvCO2 showed about a third of the increase of the paCO2 during apnoea, reaching a maximum 45 s after the onset of controlled ventilation and then being followed by a linear decrease of 0.86 mmHg/min (Fig.2b). Comparing the course of paCO2 and pvCO2 during apnoea as well as during the period of controlled ventilation, pHa and pHv changed in a reciprocal manner (Fig. 3a/b). The so-called normalization of pCO2 (paCO2 falls under pvCO2) and pH (pHa exceeds pHv) began 18.2 s and 23.2 s respectively after the onset of controlled ventilation (Fig. 4a, b). CONCLUSION. Considering the expected decrease of paO2 during hyperoxic apnoea, insufficient pulmonary N2 elimination prior to the onset of apnoea, as well as direct N2 delivery into the alveoli, due to the so-called a ventilatory mass flow, will limit unrestricted pulmonary O2 uptake. The continuing decrease of the paCO2 after the onset of controlled ventilation can be regarded as indirect proof of a ventilatory mass flow. The course of pCO2 and pH after the onset of controlled ventilation shows that normalization in arterial and mixed-venous blood gas status takes place in about 18.2 s after the cessation of apnoea. PMID- 8250204 TI - [The effect of hyperbilirubinemia on the measurement of oxygenated hemoglobin (O2Hb), carboxyhemoglobin (COHb) and methemoglobin (MetHb) using multiwavelength oximeters in mixed venous blood]. AB - Oximetric measurements are influenced by several mechanisms. Severe jaundice is one of these mechanisms with some clinical interest. In the literature it is pointed out that a high bilirubin concentration may falsify oximetric measurements and is often accompanied by elevated COHb levels. The reason for this phenomenon is thought to be an interference in the absorption spectra of haemoglobin derivatives and bilirubin [2, 3, 4, 10]. In our investigation we attempted to answer the following questions: 1. How do multiwavelength oximeters measure haemoglobin derivatives in different bilirubin concentrations? 2. Do different multiwavelength oximeters give different concentrations of haemoglobin derivatives? METHODS. In 13 patients who developed postoperative jaundice on the intensive care unit, O2Hb, COHb and MetHb were measured in mixed venous blood with two multiwavelength oximeters (OSM3, Radiometer; CO 2500, Ciba-Corning). Bilirubin concentration was measured by the DPD (dichlorphenyldiazonium) method in the central laboratory of our hospital. RESULTS. With increasing bilirubin concentrations, both oximeters measured increasing O2Hb values; the OSM3 consistently showed higher O2Hb concentrations than the CO 2500, with a maximal difference of 2.8% (Fig. 3). Regarding COHb, we saw clear increases in the values with increasing bilirubin concentrations (Fig. 4). The CO 2500 showed higher COHb values than the OSM3 (average 1.54 +/- 0.3%). The findings regarding MetHb differed. The CO 2500 showed increasing MetHb values as the bilirubin concentration increased (Fig. 5). All measurements exceeded normal values above a bilirubin concentration of 17 mg/dl. The OSM3, however, measured constant MetHb values which did not depend on jaundice. CONCLUSIONS. 1. The in vitro measurement of haemoglobin derivates by multiwavelength oximeters is influenced by hyperbilirubinaemia. This is caused by an interference between the light absorption spectra of the haemoglobin derivates and of bilirubin and by the increasing development of endogenous CO in the haem metabolism during severe jaundice (Fig. 7). 2. With increasing bilirubin levels, a lower O2Hb is measured with the CO 2500 than with the OSM3. 3. We also see increasing COHb values with rising bilirubin concentrations. 4. With increasing bilirubin levels the MetHb concentration measured with the CO 2500 rises, while the OSM3 gives constant MetHb values. 5. In severe jaundice the O2Hb values measured with multiwavelength oximeters are not identical with the real blood concentration of this haemoglobin derivative. In this situation multiwavelength oximeters cannot be used as a reference method for in vivo oximetric systems such as pulse oximeters or fibreoptic pulmonary artery catheters. PMID- 8250205 TI - [Optimization of the image intensifier-assisted technique of lumbar sympathetic block. Computed tomographic simulation of a paravertebral puncture access]. AB - The paravertebral approach is the most common technique for local anaesthetic and neurolytic lumbar sympathetic blocks. However, guidelines on the site of needle insertion differ. As there have been several case reports on accidental trauma to the ureter and the kidney, this study was undertaken to evaluate the site of paravertebral needle insertion and the fluoroscopic landmarks for lumbar sympathetic blocks by computed tomographic puncture simulation. METHODS. CT scans of 73 patients at the level of L2-4 were analysed with respect to the location of kidneys, the vertebral insertion of the diaphragm, the distance of the sympathetic trunk from the ventral and lateral border of the lumbar vertebrae, the paravertebral distance of a tangent from the sympathetic trunk through the kidney surface and the frequency of inadvertent puncture of major organs by different paravertebral approaches. For needle insertion distances of 6, 8, 10 and (at the level of L4) 12 cm lateral to the midline were simulated. RESULTS. Simulating a paravertebral approach of 6 cm resulted in perforation of the right (or left) kidney only at the level of L2 in 1.4% (2.8%) of cases. The incidence of accidental renal puncture due to a paravertebral approach of 8 cm was 26.0%, 4.1% and 0 (26.0%, 2.7% and 0) at the levels L2, L3 and L4, respectively. A more lateral insertion of needles 10 cm from the spinous process increased the frequency of anticipated renal puncture to 57.5%, 19.2% and 1.4% (65.8%, 26.0% and 1.4%); in addition, perforation of liver parenchyma was detected in two cases. Although the lower pole of the kidney reached the mid-vertebral level of L4 in only 23.3 (15.1)% of cases, a lateral approach 12 cm from the midline still showed a 8.2 (4.1)% incidence of kidney perforation and inadvertent trauma to the intestine in two cases. The mean distance from the sympathetic trunk to the ventral border of the lumbar vertebra (in simulation of a lateral fluoroscopic view) was 0.80, 0.66 and 0.59 cm, analogous measurements to the lateral border averaged 0.37, 0.43 and 0.50 cm at L2, L3 and L4, respectively. At the level of L2 the medial insertion of the diaphragm was identified in 45% of patients in close anatomical relationship to the psoas fascia. CONCLUSION. In order to reduce the risk of accidental trauma to major organs the paravertebral distance of insertion of the needles from the midline should not exceed 6, 7 and 10 cm for lumbar sympathetic blocks at the levels of L2, L3 and L4, respectively. However, a paravertebral approach of less than 6 cm may cause a lateral and ventral deviation of the needle from the sympathetic chain. Under fluoroscopy a correct needle position is obtained at an average distance of 0.5-0.8 cm dorsal to the anterior vertebral border, advancing the needle to the ventral border may cause an accidental puncture of the vena cava in more than 20% of patients undergoing nerve block of the right sympathetic chain. Furthermore, at the level of L2 inadvertent placement of the needle tip within the vertebral insertion of the diaphragm must be considered as a reason for atypical spread of contrast medium. PMID- 8250206 TI - [Anesthesia and intensive therapy for a patient with mitochondrial myopathy]. AB - Since 1983 we have been involved in the diagnostic work-up and emergency treatment of a female patient now 48 years old who has a mitochondrial myopathy resembling Luft's disease. The syndrome was first described in 1959, and in more detail in 1962, by Luft and et al., who reported a picture of hypermetabolism with high temperature, extreme sweating, tachycardia, dyspnoea at rest, polydipsia, polyphagia and irritability but normal thyroid function. In 1971 and 1976 Haydar and Di Mauro presented a second case and proposed treatment with chloramphenicol. Our patient has the third case of the syndrome reported so far: her case was initially published in 1987. CASE REPORT. Since her 17th year of life the patient had suffered from episodes of fever, tachycardia and sweating. At the age of 32 these attacks worsened, leading to unconsciousness and apnoea. The patient then had to be intubated, ventilated and sometimes resuscitated. The diagnosis of MH susceptibility and Luft's disease was made on biochemical grounds after the first muscle biopsy in 1983. Therapy with chloramphenicol failed. Therapy with beta blockers, vitamin C and K or E, coenzyme Q10 and a high-caloric diet was started in 1985. The patient was registered with an emergency service, which flew her to our ICU whenever she had a severe crisis. For milder episodes she was supplied with an oxygen breathing mask at home. Myalgia increased with the episodes starting in 1988, and the patient needed dantrolene infusions and analgesics at home. To facilitate venepuncture a Port-A-Cath system was implanted in 1987, which had to be removed four times due to infection and sepsis. A muscle biopsy was taken in Rotterdam, which revealed differences in mitochondrial function from the biochemical findings recorded in 1983 and not in keeping with Luft's disease. Unfortunately, the patient was not able to undergo further metabolic investigations or therapeutic trials. ANAESTHESIA. The patient received three local and six general anaesthetics in our clinic. The muscle biopsies, two in 1983 and one in 1985, were performed under local infiltration with procaine and were uneventful. The general anaesthetics were carried out without MH trigger substances following pretreatment with dantrolene for the following surgical procedures: the repair of an extensive arterio-venous fistula between the brachiocephalicus trunk and the right jugular and subclavian vein, revision of the sternum cerclage, implantations and explanations of infectious Port-A-Cath systems. We used etomidate, propofol and fentanyl or alfentanil with nitrous oxide and oxygen for induction and maintenance of anaesthesia. Muscle relaxation was induced with vecuronium or atracurium. All cardiovascular, respiratory, metabolic and temperature measurements stayed in normal ranges. After the extensive vascular repair (av fistula) the patient had to be mechanically ventilated for some hours until normal body temperature was restored. At the end of all other periods of anaesthesia she was extubated in the operating theatre. In five cases the postoperative period was uneventful. Only once she developed a crisis with hyperthermia, tachycardia, sweating and dyspnoea. INTENSIVE CARE. From 1985 to 1992 the patient was treated in our ICU 21 times. On 11 occasions she was already intubated and being ventilated by the emergency service on arrival. Extubation was usually possible within 2-20 h. During the crisis, heart rate was about 160-190 per minute and temperature above 40 degrees C. Serum values of CK, glucose, BUN, electrolytes, lactate and thyroid hormones were always in the normal ranges. Blood gas controls showed a constant respiratory alkalosis, arterial pCO2 values decreasing to 20 mm Hg or less. In addition to mechanical ventilation, treatment consisted in dantrolene infusions and droperidol injections, supplemented from 1989 onward with piritramide injections because of the increased severity of myalgia. In 1991 we gave propofol by PMID- 8250207 TI - [Nosocomial infections in a neurosurgical intensive care unit]. AB - The occurrence of nosocomial infections in 1017 consecutive patients seen in a neurosurgical intensive care unit (ICU), over a period of 18 months is reported. The frequency of infections is low, which may possibly be due partly to the short stay in the ICU. Close interdisciplinary cooperation is stressed as an important factor in limiting infections. BACKGROUND. The aim of this study was to analyse the nosocomial infections in a neurosurgical intensive care unit over a period of 18 months, emphasizing localization and cause of infection, in order to adapt treatment and to take preventive measures. From 15% to 27% of patients treated in ICUs acquire nosocomial infections. In Germany this means 500,000-800,000 patients a year, and the annual costs related to nosocomial infections are estimated at 1.7 billion Deutschmarks. PATIENTS AND METHODS. In all, 1017 consecutive patients were evaluated. The patients were divided into two groups, depending on the duration of treatment in the ICU: Patients who remained for less than 48 h (1017 patients) Patients who were treated for a period exceeding 48 h (314 patients) The evaluation was performed retrospectively from the medical documentation. Criteria for registration are those of the Centers for Disease Control (Atlanta 1988). When more than one infection was diagnosed, each was considered as a new infection, regardless of the bacteria involved. Among the 314 patients who were in the ICU for more than 48 h a total of 114 nosocomial infections were recorded. The frequency of infection referred to all patients treated during that time (n = 1017) was 11.2%, while the frequency among those who were treated for longer than 48 h was 36.3%. Most infections (38.6%) affected the respiratory tract, followed by infections of the urinary tract. Of the bacteria determined 56.7% were gram-negative. In this group E. coli was the most frequently found (29.8%). In the group of gram-positive bacteria, S. aureus was diagnosed in 56.3% of cases. Twelve (16%) of the infected patients died and lethality referred to all patients was 8.6%. DISCUSSION. Compared with other studies, this study revealed a low the infection rate, at 11.2%. This can be explained partly by the short stay in this ICU (mean 3.7 days) and partly by the retrospective method of registration and the particular medical characteristics of neurosurgical patients. The well-known general risk factors for infection, such as age, mechanical ventilation, continuous catheterization of the bladder, and long duration of stay, are also found in neurosurgical ICUs. It is quite difficult to determine to what extent nosocomial infections prolong the treatment necessitated by the primary neurosurgical disease. We were not able to extrapolate the influence of immunosuppressant treatment on the appearance of nosocomial infections, as almost all patients in this study were receiving steroids. This study underlines the necessity of interdisciplinary cooperation between neurosurgeons, anaesthesiologists, microbiologists and nurses in neurosurgical ICUs, where most patients staying longer than 48 h are immunosuppressed and ventilated and thereby particularly at risk of nosocomial infections. PMID- 8250208 TI - [The pulmonary artery catheter]. AB - The introduction of the pulmonary artery (PA) catheter in routine practice by Swan and Ganz at the beginning of the 1970s has enhanced our understanding of the mechanisms of patients' decompensation and has helped to guide appropriate therapeutic interventions in many diseases. Everybody who uses this monitoring instrument should become familiar with its technical concepts and theoretical basis. Modern technology has provided a large number of sophisticated developments, particularly in the past decade. Today we are able to obtain a huge mass of haemodynamic and oxymetric data in the critically ill. It is a common misconception in clinical practice that PA catheters should be employed for documentation of the haemodynamic catastrophe; however, it should be used to avoid significant haemodynamic problems. In recent years the value of PA catheter monitoring has been called into question. Whether a monitoring device can improve 'outcome' may be doubted. Although some opponents of haemodynamic monitoring wish to turn back the clock to the old days of a 'finger on the pulse', there is no doubt that the use of the PA catheter may yield additional information or that some of this information may be useful when the data are interpreted carefully. The time has come not only to criticize this monitoring instrument, but to examine future perspectives and visions. PMID- 8250209 TI - Electroinsertion of proteins in the plasma membrane of red blood cells. PMID- 8250210 TI - Determination of integrins on cells by cell adhesion to antibodies. AB - An assay for the determination of relative concentrations of integrins on clonal cultured cells is described. Unpurified monoclonal antibodies to integrin subunits, present in ascites or hybridoma culture media, are immobilized on a plastic surface via goat antibodies to mouse IgG. Cell attachment to the integrin coated substrate is quantitated after fixation and staining of the cells. The assay is specific and very sensitive; only cells with the relevant integrins attach to antibody-coated substrates and concentrations of antibody as low as 1 10 ng/ml are sufficient. Titrations of the antibodies on the solid phase allow the estimation of the relative amounts of integrins on different cell types. The results with this assay correlate well with results obtained by flow cytometry. PMID- 8250211 TI - Capsular polysaccharide structure of a clinical isolate of Vibrio vulnificus strain BO62316 determined by heteronuclear NMR spectroscopy and high-performance anion-exchange chromatography. AB - Virulence of Vibrio vulnificus has been strongly associated with encapsulation. Capsular polysaccharide was purified from a virulent strain of Vibrio vulnificus BO62316, a clinical isolate, by dialysis, centrifugation, enzymatic digestion, and phenol-chloroform extraction. This polysaccharide shows partial reactivity with antibodies to the capsular polysaccharide of a related pathogenic strain of V. vulnificus (MO6-24) whose structure was recently reported. Nuclear magnetic resonance spectroscopic analysis of the purified polysaccharide from strain BO62316 showed that the polymer is composed of a repeating structure with four sugar residues per subunit: a residue of 2-acetamido-2,6-dideoxy-hexo pyranose in the alpha-gluco configuration (QuiNAc), a residue of 2-acetamido-2,6-dideoxy hexopyranose in the alpha-galacto configuration (FucNAc), a residue of 2 acetamido-2,6-dideoxy hexopyranose in the alpha-manno configuration (RhaNAc), and a residue of 2-acetamido-2,6-dideoxy hexouronate in the alpha-galacto configuration (GalNAcA). The complete carbohydrate structure of the capsular polysaccharide was determined by heteronuclear nuclear magnetic resonance spectroscopy and by high-performance anion-exchange chromatography. The 1H and 13C spectra were completely assigned, and vicinal coupling relationships were used to establish the stereochemistry of each sugar residue, its anomeric configuration, and the position of the glycosidic linkages. The complete structure is Formular; [see text]. This is the first reported occurrence of RhaNAc in a capsular polysaccharide. PMID- 8250212 TI - An examination of the reliability of the radiochemical assay for monoamine oxidases A and B. AB - The radiochemical assay for MAO A has been compared with the polarographic and alcohol dehydrogenase-coupled assays, using 5-hydroxytryptamine (5-HT) and tyramine and the homogeneous human liver enzyme expressed in yeast, and rat liver mitochondria. Despite efforts to measure true initial rates and to avoid known sources of error in the radiochemical procedure, significantly higher rates (Vmax) and lower Km values for the substrate were obtained with the polarographic than with the radiochemical method for 5-HT and tyramine, using either highly purified enzyme or mitochondria. The rate of tyramine oxidation measured by the polarographic method and the coupled assay agreed well, however. Consequently, for kinetic and inhibition studies we recommend the polarographic method for MAO A substrates. Comparison of the radiochemical, polarographic, spectrophotometric, and coupled assays for MAO B from beef liver (mitochondria and homogeneous enzyme) showed polarography to be the method of choice on all substrates tested, although, if all known sources of error are carefully controlled, the radiochemical assay gives the same Vmax and Km values. We also report that with each substrate studied the apparent Km is much lower when mitochondria are used than when the highly purified, virtually lipid-free preparations of MAO A are studied. PMID- 8250213 TI - A spectrophotometric assay for alpha-ketoaldehydes using horse liver alcohol dehydrogenase. AB - alpha-Ketoaldehydes have been extensively employed as reagents for the chemical modification of arginine residues in proteins, and to probe for putative anion recognition sites. A major disadvantage in their use is instability, as alpha ketoaldehydes are prone to hydration, oxidation, and polymerization. These reagents are typically supplied as the aldehyde hydrates which are relatively stable, but must be purified and standardized prior to analytical use. The known chemical, spectroscopic, and enzymatic methods for quantitating alpha ketoaldehydes are not practical for routine analysis due principally to their high detection limits and sensitivity to interfering substances. Surprisingly, alpha-ketoaldehydes have not been reported as substrates for the alcohol dehydrogenases. We have discovered that phenylglyoxal and several related alpha ketoaldehydes are good substrates for horse liver alcohol dehydrogenase (HLADH). The second order rate constants (kcat/Km) are within a factor of 10 of that for the reduction of acetaldehyde, a known good substrate for HLADH. The enzymatic reduction reaction is stoichiometric with the oxidation of NADH, resulting in a rapid, convenient, and sensitive method for the spectrophotometric quantitation of alpha-ketoaldehydes, with a submicromolar detection limit for phenylglyoxal. The sole product of phenylglyoxal reduction has been identified as alpha hydroxyacetophenone. The ketone functional group is not reduced, and the enzymatic reaction is essentially irreversible as alpha-hydroxyacetophenone is not oxidized to phenylglyoxal by HLADH in the presence of NAD+. PMID- 8250214 TI - The use of tributylphosphine and 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole in the study of protein sulfhydryls and disulfides. AB - The use of the reagent tributyl phosphine (Bu3P) to reduce disulfides (Ruegg, U.T., and Rudinger, J., Methods Enzymol. 47, 111-116, 1977) and of 4 (aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (ABD-F) to block free sulphydryl groups (Toyo'oka, T., and Imai, K. Anal. Chem. 56, 2461-2464, 1984) is well established in the literature. Since the two reagents apparently do not react with each other, their combination offers a convenient and quite general method for the complete characterization of free Cys (SH) and crosslinked Cys (S-S) in proteins (Kirley, T.L., J. Biol. Chem. 264, 7185-7192, 1989). We review some of the characteristics of the reaction of these reagents with Cys in peptides and proteins and some of the properties of the ABD-Cys derivatives. The review includes reactions with model compounds (e.g., Cys and glutathione), proteins such as enolase from yeast and rabbit muscle, containing only free Cys, a protein, fetuin, containing only crosslinked Cys, and a protein, superoxide dismutase, containing both free and crosslinked Cys. In all cases the direct comparison of the tryptic or chymotryptic peptides derived from the products of parallel reactions of the protein with ABD-F alone and with ABD-F together with Bu3P permitted the determination of both free and total Cys in the protein. Sequencing the fluorescent ABD-peptides established the position of the Cys residues in the primary sequence.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250215 TI - On the synthesis and characterization of N-formyl glutathione and N-acetyl glutathione. AB - Treatment of glutathione (GSH) with formic acid and acetic anhydride leads to preparation of crystalline N-formyl GSH. This is the first characterized preparation of N-formyl GSH; this product was previously erroneously thought to be N-acetyl GSH. Authentic N-acetyl GSH (crystalline) was prepared by treating GSH with acetic acid and acetic anhydride. The N-acylated derivatives were characterized by 1H NMR and mass spectroscopy. PMID- 8250216 TI - Quantification of subnanomolar amounts of phosphate bound to seryl and threonyl residues in phosphoproteins using alkaline hydrolysis and malachite green. AB - An assay for protein-bound phosphate with a capacity to determine 100 pmol of phosphate is described. It is based on the combination of two well known methods: the alkaline hydrolysis of phosphate from seryl and threonyl residues in phosphoproteins and the quantification of the released phosphate by the use of malachite green and phosphomolybdate. PMID- 8250217 TI - Purification of membrane proteins using a micropreparative gel electrophoresis apparatus: purification of subunits of the integral membrane protein Bacillus subtilis aa3-type quinol oxidase for low level amino acid sequence analysis. AB - We have developed a continuous elution micropreparative gel electrophoresis system in which small amounts of proteins, e.g., 1-10 micrograms, are continuously eluted from one-dimensional polyacrylamide gel columns in either native or denatured form. Depending on the electrophoretical parameters proteins are separated according to their size and/or net charge. The system is based on a simple modification of the Mini-Protean II 2D unit from Bio-Rad Inc. The apparatus is connected on-line to a high-performance liquid chromatograph allowing accurate delivery of the elution solvent and direct analysis of the separated samples by uv detection. The high resolution of the apparatus allows the purification of individual proteins, even from complex mixtures, in only one step, with overall recoveries of approximately 70%. In addition to standard proteins the system has been applied to the purification of the membrane-bound and thus highly hydrophobic subunits I and II from Bacillus subtilis aa3-type quinol oxidase. The hydrophobic character of these subunits has hindered their purification by other conventional methods. Isolated subunits were collected directly into a buffer suitable for proteolytic digestion and automated amino acid sequence analysis. The internal amino acid sequences determined could all be found in the DNA sequence recently reported by Santana et al. (J. Biol. Chem. 267, 10225-10231, 1992), thereby confirming the expression of such an oxidase. PMID- 8250218 TI - In situ determination of the reduction levels of cytochromes b and c in growing bacteria: a case study with N2-fixing Azorhizobium caulinodans. AB - The determination of the in situ reduction levels of cytochromes b and c in growing bacteria is achieved by coupling a chemostat with a dual wavelength spectrophotometer. Visible light absorption spectra of cytochromes present in bacterial cells actively growing in a chemostat at a specific growth rate of 0.1 h-1 are recorded. This is accomplished by transporting the emitted light from the spectrophotometer via glass fibers to one side of the chemostat vessel and detecting the transmitted light via a photomultiplier at the other side. The vessel itself is enclosed in a dark box, which contains mirrors on the inside surfaces. The reduction levels of cytochromes b and c during steady state in chemostat cultures are expressed as percentage absorbance of fully reduced cytochromes in the alpha-region of the spectrum. Steady state spectra are recorded in N2-fixing, succinate-limited continuous cultures of Azorhizobium caulinodans at dissolved oxygen tensions in the range between 0.1 and 3.5% O2. Spectra of fully reduced cytochromes are obtained on the basis of spectra recorded after having reached anoxic conditions by sparging pure nitrogen gas through the culture. These spectra of cytochromes b and c reduced by endogenous substrates are corrected as to give the spectrum of fully reduced cytochromes. The respective contributions of cytochromes b and c to spectra in the alpha region are estimated by deconvolution using best-fit analysis. Using this in situ technique it is observed that at each dissolved oxygen tension the reduction level of the cytochromes b is higher than that of the cytochromes c.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250219 TI - Preparative mini-slab gel continuous elution electrophoresis: application for the separation of two isoforms of rat hepatocyte cell adhesion molecule, cell-CAM 105, and its associated proteins. AB - A simple method for performing continuous elution electrophoresis using a mini slab-gel system has been developed. The high resolving power of this method provides a means to separate complex mixtures of proteins and recover individual peptides in high yields. Recoveries between 89 and 99% were achieved using a single percentage gel to separate a mixture of standard proteins radiolabeled with 125I. Peptide fragments between 2.5 and 60 kDa generated by CNBr digestion of albumin were isolated within 150 min. Application of continuous elution electrophoresis to the purification of proteins associated with the rat hepatocyte cell adhesion molecule, cell-CAM 105, resulted in complete separation of the two predominant isoforms and the purification of three immunologically unrelated proteins that were coisolated during immunoaffinity chromatography. PMID- 8250220 TI - Facile preparation and structural determination of monohydroxy derivatives of docosahexaenoic acid (HDoHE) by alpha-tocopherol-directed autoxidation. AB - Polyunsaturated fatty acids are oxidized through both enzymatic and nonenzymatic reactions into hydroxy derivatives. With increasing interest in dietary manipulations through ingestion of the highly unsaturated fish oil fatty acids, eicosapentaenoic acid and docosahexaenoic acid (DHA), methods to measure their metabolism are required. In this study we report the simple and expedient alpha tocopherol-directed autoxidative preparation of a series of monohydroxy derivatives of DHA to provide a relatively homogeneous hydroxylation along each of the double bonds of the fatty substrate. Products were purified by high performance liquid chromatography (HPLC) and their structures elucidated by the characteristic fragmentation pattern of the hydrogenated methyl ester trimethylsilyl ether derivatives by gas chromatography-mass spectrometry. Nine products were isolated in 20.2% yield overall, ranging from 1.55 to 4.14% yield of isolated compound. These were identified as 7, 8, 10, 11, 13, 14, 16, 17, and 20-HDoHEs (monohydroxydocosahexaenoic acids). Two of these products (14- and 17 HDoHE) could not be separated under the HPLC conditions used but were clearly distinguished using selected ion chromatography by their distinct mass spectral fragmentation. This method is highly suitable for the generation of standards to investigate the metabolism of DHA in tissues. PMID- 8250221 TI - Circular dichroic analysis of denatured proteins: inclusion of denatured proteins in the reference set. AB - We hypothesize that inclusion of denatured proteins in the set of reference native proteins may better represent the unordered form in the current circular dichroism (CD) analyses of proteins involving unfolding ones. Adding three denatured-protein spectra and one oligopeptide spectrum to 16 reference protein spectra markedly improved the correlation coefficients (r) between CD calculations and X-ray determinations for the unordered form and, to a lesser extent, for beta-turn, but the r-values for alpha-helix and beta-sheet decreased slightly. With 20 reference proteins the estimates of the unordered form of denatured proteins were significantly improved. Thus, we suggest that as a compromise the new set of reference proteins be used for estimating the changes in conformation for unfolding proteins. However, the current use of 16 reference native proteins appears to be adequate for CD analysis of native proteins and the expansion to 20 reference proteins including denatured ones may not enhance the analysis of native proteins. PMID- 8250222 TI - Analysis of receptor-stimulated and basal guanine nucleotide binding to membrane G proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - A method to study [alpha-32P]GTP binding to the alpha subunit of GTP-binding proteins in rat brain membranes is described. This method measures receptor stimulated GTP binding to individual alpha subunits. GTP binding is associated with two protein bands following sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The bands, 40- and 45-kDa in size, comigrate with the alpha subunits of Gi/Go and Gs, respectively. Binding of [alpha-32P]GTP is saturable and Mg(2+)-dependent. Nucleotides compete with [alpha-32P]GTP binding in the following order: GTP > GDP > Gpp(NH)p > App(NH)p. Dopamine stimulates [alpha-32P]GTP labeling of the 40- and 45-kDa bands. A binding increase of 300 400% is observed at 10 microM dopamine. Isoproterenol (10 microM) stimulates [alpha-32P]GTP binding only to the 45-kDa protein band. The effects of dopamine and isoproterenol are blocked by their respective receptor antagonists, fluphenazine and propranolol. The individual G proteins activated by dopamine are resolved by immunoprecipitation of stimulated [alpha-32P]GTP binding to G alpha s, G alpha i, and G alpha o with specific anti-G alpha antisera. Dopamine stimulates [alpha-32P]GTP binding to G alpha s and G alpha i while the labeling of G alpha o was not significantly changed. Pertussis toxin-mediated ADP ribosylation prevents the activation of G alpha i which is mediated by dopamine receptor stimulation. The methods described are useful in defining the coupling of specific neurotransmitter receptors to specific G proteins in native membranes. These procedures also allow measurements of receptor stimulation of individual G proteins in intact biological membranes. PMID- 8250223 TI - Tetrazolium-based cell bioassay for neurotoxins active on voltage-sensitive sodium channels: semiautomated assay for saxitoxins, brevetoxins, and ciguatoxins. AB - In the present study we have developed an assay for the detection of sodium channel-specific marine toxins based upon mitochondrial dehydrogenase activity in the presence of veratridine and ouabain. This cell bioassay allows detection of either sodium channel enhancers, such as the brevetoxins and the ciguatoxins, or sodium channel blocking agents, such as the saxitoxins. The assay responds in a dose dependent manner and differentiates the toxic activity as either sodium channel blocking or enhancing. In addition, the assay is highly sensitive, with present detection limits of 2 ng/ml for either saxitoxins or brevetoxins (PbTx-1 and PbTx-3). Assay response to a ciguatoxic extract and to brevetoxins is rapid, allowing dose dependent detection within 4 to 6 h. The method is simple, utilizes readily available reagents, uses substantially less sample than required for mouse bioassay, and is well within the scope of even modest tissue culture facilities. This cell-based protocol has the potential to serve as an alternate and complementary method to the standard mouse bioassay. PMID- 8250224 TI - Isolation of the phagocytic compartment from macrophages using a paramagnetic, particulate ligand. AB - Isolation of Fc receptor-dependent phagosomes from a macrophage cell line, J774 A.1, was accomplished using antibody-opsonized, paramagnetic beads. Following binding and ingestion of these beads, cells were homogenized in a standard membrane isolation buffer. Phagosomes containing the trapped paramagnetic beads were isolated by subjecting the whole cell homogenate to a magnetic field. The method is extremely simple and the preparation of an enriched phagosome fraction from a whole cell homogenate is rapid and highly selective. The method should provide useful starting material for investigators interested in cytoskeletal involvement in phagocytosis, in kinetic studies of ingestion, in phagosome lysosome fusion, and in the ability of a particular ligand to initiate phagocytosis. PMID- 8250225 TI - Isolation and characterization of the outer membrane proteins of Serratia marcescens W225. AB - The study addressed the general problem of fractionating cell envelopes in order to isolate the outer membranes of gram-negative bacteria. Whereas the cells are normally transformed into spheroplasts prior to disintegration and membrane separation, Serratia marcescens was found to be resistant to spheroplast formation using the procedures available, which were originally developed for Escherichia coli. An efficient technique for spheroplasting S. marcescens was therefore developed; this comprised combining osmotic shock and lysozyme-EDTA treatment of sucrose-conditioned cells. Spheroplasting efficiency and the amount of outer membrane protein recovered were highly dependent on the spheroplasting technique used. Separation of the outer and inner membranes was performed by two methods, isopyenic centrifugation and selective detergent solubilization with Sarkosyl. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the analysis of specific inner membrane marker enzymes revealed that the protein obtained by detergent solubilization was much purer than that obtained by isopycnic centrifugation. The outer membrane isolated accounted for 60% of the envelope proteins and had a buoyant density of 1.2502 g/cm3. The protein profile of the outer membrane determined by SDS-PAGE resolved into 12 distinct protein bands, 3 of which represented major proteins. PMID- 8250226 TI - Synthesis of N alpha-[3H]acetyl-L-lysine chloromethyl ketone and its use in the fluorographic detection of proteases. AB - Tritiated N alpha-acetyl-L-lysine chloromethyl ketone (ALCK) was synthesized on a laboratory scale for use as an active-site-directed affinity label in the fluorographic detection of proteases after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The synthesis involved acetylation of N epsilon benzyloxycarbonyl-L-lysine chloromethyl ketone with [3H]acetic anhydride just before the removal of the benzyloxycarbonyl group. By this method, [3H]ALCK with a specific activity of 250 mCi/mmol was obtained as a crystal. Trypsin, thrombin, plasmin, papain, and clostripain were inactivated by ALCK according to first order kinetics. For fluorographic detection of proteases, enzyme samples were allowed to react with [3H]ALCK and then resolved by SDS-PAGE. Proteases that reacted with [3H]ALCK could be detected with a sensitivity equivalent to or higher than that of Coomassie brilliant blue R-250 staining. A trypsin-like protease in Pronase, clostripain as a contaminant in a commercial preparation of Clostridium histolyticum collagenase, and cysteine proteases in Porphyromonas gingivalis could be detected. PMID- 8250227 TI - Preparation and characterization of biotinylated analogs of the calcium channel blocker omega-conotoxin. AB - We have prepared a series of biotinylated analogs of omega-conotoxin (omega CgTx) as potent, selective markers for N-type calcium channels. At pH 9.5, reaction of omega CgTx with amidocaproylbiotin succinimidyl ester gives three biotinylated conjugates, labeled at lysines 2 or 24, or at both positions. Kinetic competition assays of 125I-omega CgTx binding to rat brain synaptic membranes show that each conjugate has a similar rate constant for association (1-1.3 x 10(6) M-1 s-1) but not dissociation (1-4 x 10(-4) s-1). Comparison with rate constants obtained for the association (1.2 x 10(7) M-1 s-1) and dissociation (5 x 10(-5) s-1) of native omega CgTx indicates that while biotinylation reduces omega CgTx potency (Kdkin = k-2/k2 = 4 pM for omega CgTx), binding of these labels to membranes is nevertheless of very high affinity (Kdkin 0.1-0.3 nM). PMID- 8250228 TI - An amperometric flow-injection analysis biosensor for glucose based on graphite paste modified with tetracyanoquinodimethane. AB - A biosensor system using flow injection analysis (FIA) has been developed for the analysis of glucose in human serum. The system consists of the enzyme glucose oxidase incorporated into graphite paste modified with the electroactive material tetracyanoquinodimethane (TCNQ). TCNQ acts as an efficient mediator for oxidation of the reduced enzyme at 200 mV vs Ag/AgCl. The flow injection assay described has detection limits of 2 mM glucose using a 100-microliters sample injection through a 250-microliters sample loop. Data are presented to show the effect of sample injection volume and flow rate on the response of the FIA sensor. The biosensor exhibited excellent reproducibility for 800 injections. The loss of response after 800 injections was due to leaching of TCNQ from the graphite paste. Each assay takes 3 min giving a sample throughput of 20 per hour at a flow rate of 30 ml/h. The sensor was applied to the determination of glucose in human serum. The glucose measurements are in good agreement with those of a commercially available spectrophotometric method. Data showing the effect of interfering substances, ascorbic acid and acetaminophen, on the response of the sensor are also reported. PMID- 8250229 TI - Analysis of lipids in crude extracts by 13C nuclear magnetic resonance. AB - Phospho- and glycolipids with mono- or polyunsaturated fatty acid chains, free and acylated steroids, carotenes and carotenoids, chlorophylls, and related pheophytins were detected in crude extracts by NMR spectroscopy without any need for prior separation. A broad range of molecules belonging to different lipid classes could be identified in one-dimensional 13C NMR spectra obtained by applying a polarization transfer pulse sequence (DEPT 135 degrees). Directly detected or 1H-detected two-dimensional heterocorrelated NMR experiments were performed to facilitate the assignment of peaks arising from carotenoids, unsaturated fatty acid chains, and chlorophylls. 13C NMR data of crude lipid extracts from the macroalagae Ulva rigida and Fucus virsoides are shown to yield an informative overview of their lipid content. NMR is thus proposed as a simple, nonselective, and nondestructive technique for the first screening of the main lipid classes in complex lipid mixtures. PMID- 8250230 TI - Lactose repressor-operator DNA interactions: kinetic analysis by a surface plasmon resonance biosensor. AB - Lactose repressor binding to operator DNA and subsequent dissociation of the complex was monitored continuously by a biosensor, measuring surface plasmon resonance. In this analysis a synthetic, double-stranded oligonucleotide containing the operator site was immobilized on the sensor surface and repressor protein was passed over the surface. The formation of the repressor-operator complex was specific and could be inhibited by isopropyl-beta-D thiogalactopyranoside inducer. From the association curve, the apparent kass was determined to be 1.8 x 10(6) M-1 s-1. Dissociation of the complex was, for the first time for the lac repressor, determined as an uncatalyzed reaction and the kdiss was determined to be 3.4 x 10(-4) s-1. As a reference, the repressor operator interaction was analyzed by electrophoretic mobility shift assay under similar reaction conditions. With this method the equilibrium binding constant was calculated to be 2.4 (+/- 0.2) x 10(8) M-1. The corresponding value calculated from biosensor data was 5.1 x 10(9) M-1. PMID- 8250231 TI - Determination of the antimetabolite Gemcitabine (2',2'-difluoro-2'-deoxycytidine) and of 2',2'-difluoro-2'-deoxyuridine by 19F nuclear magnetic resonance spectroscopy. AB - The analysis of the new antimetabolite 2',2'-difluoro-2'-deoxycytidine (Gemcitabine, Eli Lilly Corp.) and of its metabolic deamination product, 2',2' difluoro-2'-deoxyuridine, in urine and plasma by means of 19F NMR is described. Both compounds show AB-type NMR spectra which are characterized as a function of pH at a magnetic field strength of 9.4 T. In the physiological pH range the spectra of both compounds are distinct, despite the fact that both compounds have almost identical average 19F shifts. A linear relation between NMR intensity and concentration in urine has been established. The NMR method does not require sample pretreatment and allows for rapid, nondestructive analysis of Gemcitabine concentrations over 0.01 mM. Separation of the two compounds in vivo by 19F NMR spectroscopy is difficult to achieve. PMID- 8250232 TI - A generally applicable ELISA for the detection and quantitation of the cytosolic factors of NADPH-oxidase activation in neutrophils. AB - An enzyme-linked immunosorbent assay (ELISA) has been developed using polyclonal antibodies raised against two cytosolic proteins of 47 kDa (p47) and 67 kDa (p67) which behave as activation factors for the superoxide-generating NADPH oxidase of neutrophils at the onset of phagocytosis. These two proteins become associated with the NADPH oxidase complex during activation. This immunological technique has been used to follow the purification steps of p47 and p67. It allows the detection of very small amounts of cytosolic factors in a crude neutrophil extract. It is straightforward and much more sensitive (about 1000 times more) than the classical assay based on the use of a cell-free system of oxidase activation and production of superoxide anion. The percentages of p47 and p67 assessed by ELISA with respect to total cytosolic protein were estimated to amount to 0.13 and 0.20%, respectively. The described method has potential applications for the titration of the cytosolic factors of oxidase activation in autosomal recessive forms of chronic granulomatous disease. PMID- 8250233 TI - Determination of metabolites by 1H NMR and GC: analysis for organic osmolytes in crude tissue extracts. AB - World-wide salinity and drought problems necessitate the understanding of biological adaptation to water deficit. Osmotic adjustment via organic solutes is a common strategy for organisms to deal with water deficit problems. Numerous water-soluble organic metabolites across several chemical classes are commonly utilized as osmolytes, including betaines, sulfonium and sulfonate compounds, amino acids, carbohydrates, and polyols. To deal with the complexity and variability in osmolyte composition, we have devised an analytical approach that combines high-resolution 1H NMR and GLC to provide both structure identification and quantification of a broad spectrum of compounds. This combined approach also facilitated direct analyses of crude tissue extracts without extensive sample preparation, making it well-suited for a convenient screening of potential osmolytes. The structures of known osmolytes were confirmed from two-dimensional total correlation 1H NMR spectra, which also yielded structural information about unknown compounds. Five each terrestrial plant and marine animal species were examined for 41 metabolites, including osmolyte candidates glycinebetaine, dimethylsulfoniopropionate, taurine, proline, glycine, asparagine, alanine, glutamine, glucose, and sucrose. The osmotic function of glycinebetaine, proline, asparagine, glutamine, glucose, and sucrose was also demonstrated in leaves of Distichlis spicata under different salinity treatments. PMID- 8250234 TI - A fluorometric microplate-based assay of submicrogram monomeric actin by inhibition of deoxyribonuclease I. AB - Inhibition of bovine pancreatic deoxyribonuclease I (DNase I) has been a standard method for specific quantitation of monomeric (G-) actin. The aim of this work is to substantially enhance the sensitivity of this type of G-actin assay by using a fluorescent dye, thiazole orange (TO), which has a high optical absorption, moderate DNA affinity, and large fluorescence enhancement upon binding to DNA. The high fluorescence output and moderate affinity of TO's DNA complex produce a large fluorescence decrease when the complex is disrupted by DNase I, thus permitting a highly sensitive detection of G-actin by its inhibition on the fluorescence decrease. The results show that the dynamic responsiveness of a fluorometric G-actin assay can be improved by lowering the concentration of DNA and affinity of the labeling dye, as long as the fluorescence signal of the dye DNA complex is sufficient for instrumental detection. With a DNA concentration of 6.0 micrograms/ml and TO dye labeling, as little as 20 ng of G-actin can be reliably detected in a fluorescence microplate scanner. This sensitivity also appears to be the utmost detection limit of any G-actin assay that is based on DNase I inhibition. PMID- 8250235 TI - Simultaneous determination of creatine compounds and adenine nucleotides in myocardial tissue by high-performance liquid chromatography. AB - A rapid high-performance liquid chromatography method was developed for the determination of creatine phosphate, creatine, adenine nucleotides, and related compounds in myocardial tissue. Analysis was performed by reversed-phase chromatography on a C18 column containing 3-microns particles, employing gradient elution and uv detection at 210 nm. Separation was achieved in less than 5 min. Total analysis time, including equilibration of the column after return of the gradient to starting conditions, was 8 min. The high reproducibility and short analysis time make this method suitable for the routine analysis of large series of samples. PMID- 8250236 TI - Quantitative slot-blot chemiluminescence assay for determination of myeloperoxidase from human granulocytes. AB - Using slot blot, we show that myeloperoxidase (MPO), a constituent of azurophil granules of neutrophil polymorphonuclear leukocytes, can be measured quantitatively using a commercially available chemiluminescence kit, originally developed for detection of specific proteins on Western blots. MPO is determined through its ability to catalyze the oxidation of luminol, resulting in the emission of light which is recorded on a photographic film. The sensitivity of the method is high and allows MPO from less than 100 cells to be detected. This method was used to determine MPO in exudate fluid and in neutrophil fractions following disintegration and subcellular fractionation of the postnuclear supernatant on two-layer Percoll gradients. PMID- 8250237 TI - Synthesis of preparative amounts of biologically active interleukin-6 using a continuous-flow cell-free translation system. AB - Cell-free synthesis of preparative amounts of interleukin-6 (IL-6) in a wheat germ extract is described. Following transcription of uncapped mRNA by SP6 polymerase, protein was synthesized in a continuous flow of translation components. The system remained active for at least 50 h, producing 1.25 x 10(6) units of IL-6. Recovered IL-6 was > 80% pure, reacted with anti-IL-6 polyclonal antiserum in Western blots, and had a specific activity comparable to recombinant IL-6 purified from Escherichia coli. This is the first time biologically active IL-6 has been prepared in preparative amounts using a continuous-flow cell-free translation system and confirms the feasibility of using enhanced in vitro translation for rapid synthesis of proteins from cloned templates. PMID- 8250238 TI - A quantitative immunostaining method for the measurement of UDP galactose:lactosylceramide galactosyltransferase for the synthesis of globotriaosylceramide in rabbit small intestine and HeLa cells. AB - Galactosyltransferase is required for the addition of galactose to lactosylceramide (galactose beta 1-4 glucose beta 1-1 ceramide), resulting in the synthesis of globotriaosylceramide (Gb3). We describe a quantitative more sensitive and specific method for the measurement of UDP galactose:lactosylceramide galactosyltransferase activities in rabbit small intestine and HeLa cell which utilizes the specific binding of Shiga toxin to the product, Gb3. Intestinal microsomal or HeLa cell sonicate preparations were incubated in the presence of lactosylceramide and [14C]UDP-galactose. The lipid reaction products were extracted on C18 Bond-Elut columns, separated by high performance thin-layer chromatography and exposed to Shiga toxin followed by polyclonal rabbit anti-Shiga toxin antibody and goat anti-rabbit IgG alkaline phosphatase conjugate. Gb3 was visualized with NBT and BCIP and quantitated by densitometry. These data were compared with a standard assay in which, following incubation and lipid extraction, radioactivity was measured by scintillation counting of the isolated lipids. There was a 22-fold increase in enzyme activity by the immunostaining method compared to the usual scintillation counting technique. This is attributable to the exclusion of radioactive lipids other than Gb3 in calculating enzyme activity and the correction for endogenous UDP galactose. Thus, the immunostaining method provides increased accuracy, sensitivity, and specificity in the assay of galactosyltransferase activity. PMID- 8250239 TI - In vivo analysis of glucose-induced fast changes in yeast adenine nucleotide pool applying a rapid sampling technique. AB - Transition from glucose limitation to glucose excess in Saccharomyces cerevisiae leads to a fast change from complete to partial oxidative metabolism. To determine rapid changes of in vivo concentrations of yeast metabolites, occurring in the range of seconds after a glucose injection, a fast sampling technique was developed. A harvesting device with negligible dead space and highly efficient inactivation and extraction steps was applied to analyze the behavior of the physiological concentrations of adenine nucleotides as the dynamic response to fast changing glucose concentrations. Adenine nucleotides were selected as appropriate references due to their high turnover rates (in the range of seconds). During the first 5 s after a glucose pulse to a continuous chemostat culture, a significant decrease (30%) in ATP concentration and a coincident rise of ADP and AMP values were found. A further advantage of this technique is the high sampling frequency (5 s) and the long-term sterility ensuring recurrent harvesting. PMID- 8250240 TI - Testing potential gyrase inhibitors of bacterial DNA gyrase: a comparison of the supercoiling inhibition assay and "cleavable complex" assay. AB - Inhibitory activity of test compounds against Escherichia coli DNA gyrase in a "cleavable complex" assay, readily observed in vitro at the enzyme level by the artificial addition of a denaturing agent, is found to be an excellent indicator of 4-quinolone inhibition of DNA gyrase, and as accurate a predictor of target enzyme inhibitory activity as the measurement of the inhibition of DNA gyrase supercoiling. This study was designed to examine the specificity of DNA gyrase inhibitors of various chemical classes in these two DNA gyrase assays, and define the use of these two assays in understanding the nature of inhibition by experimental compounds. Supercoiling inhibition was detected by determination of the 50% inhibition level, and cleavable complex inhibition measured by the determination of the drug concentration at which 50% of the maximal (of control) formation of linear, cleaved DNA was obtained. Results indicate that these two assays can serve several different functions in microbiological research, among them: (1) quantitation of enzyme inhibitory activity at the target level; and (2) distinguishing between nonspecific inhibition or artifactual inhibition of DNA gyrase and true, mechanism-based inhibition of the catalytic activity of DNA gyrase. PMID- 8250241 TI - A rapid method for the synthesis of methylmalonyl-coenzyme A and other CoA esters. AB - A rapid and high-yield synthesis of methylmalonyl coenzyme A is reported. The two step procedure involves preparation of the thiophenyl ester of methylmalonic acid using dicyclohexylcarbodiimide as a condensing agent, followed by transesterification with coenzyme A, to yield methylmalonyl coenzyme A in 80% overall yield. Inclusion of an additional step, methylation of malonic acid with iodomethane, affords the opportunity for introducing a stable or radioactive isotope into the product. This method should be applicable for the syntheses of other coenzyme A esters that are of biochemical interest such as succinyl coenzyme A. PMID- 8250242 TI - Anion exchange separation and pulsed amperometric detection of inositols from flower petals. AB - A rapid and specific method for the separation of cyclic sugar alcohols by anion exchange HPLC and subsequent detection by pulsed amperometry is presented. The cyclitols include the inositols and O-methyl inositols. Although examples use plant extracts the method has general application and is suitable for detecting as little as 10 pmol of substance. Carbohydrates are weak acids with pKa values typically above 12. While separation of most monosaccharides on high-performance anion exchange columns in aqueous NaOH is adequate, the ionization of inositols and O-methyl inositols is weak and their separation is poor. Use of a high capacity anion exchange column separated the inositols well but the more ionized sugars were tightly retained. Automatic switching of the inositol fraction eluting from the lower-capacity column onto the high-capacity column with subsequent detection by pulsed amperometry gave specific and rapid separations of the inositols with a turnaround time of 16 min. As an example of application, we chose the separation and identification of cyclic and acyclic sugar alcohols in flower petals from a variety of plants. There was an apparent correlation between the postharvest longevity of flowers and the content of inositols, particularly D pinitol. PMID- 8250243 TI - Quantification of type III iodothyronine deiodinase activity using thin-layer chromatography and phosphor screen autoradiography. PMID- 8250244 TI - Restoring free sulfhydryl groups in synthetic peptides. PMID- 8250245 TI - Modification of a nonincinerative method for determination of iodine in iodoproteins. PMID- 8250246 TI - Enhanced transformation of Streptococcus mutans by modifications in culture conditions. PMID- 8250247 TI - A fluorescence-based protein assay for use with a microplate reader. PMID- 8250248 TI - Automatic microinjector for amphibian oocytes, eggs, and embryonic blastomeres. PMID- 8250249 TI - Functional analysis of mutated cDNA clones by direct use of PCR products in in vitro transcription/translation reactions. PMID- 8250250 TI - Detection of biotinylated molecules in solid-phase assays using a recombinant biotin-binding bacteriophage. PMID- 8250251 TI - A phosphodiesterase assay using alumina microcolumns. PMID- 8250252 TI - Analysis of the transformation products of dehydro-L-ascorbic acid by ion-pairing high-performance liquid chromatography. AB - Dehydro-L-ascorbic acid, the oxidation form of L-ascorbic acid, converts nonenzymatically to a variety of transformation products depending upon the incubating conditions. These products, called the ascorbate cascade, possess different absorption spectra and reducing activities. Ion-pairing high performance liquid chromatography with multiwavelength uv absorption and amperometric electrochemical detection has proven to be a useful technique for the analysis of these products. Separation and characterization in the present investigations reveal three kinds of transformation products: the polymerized intermediates of browning reaction, the reductant possessing an absorption maximum at 345 nm as the six-carbon compound, and erythro-L-ascorbic acid, 2 furoic acid, 3-hydroxy-2-pyrone, 5-methyl-3,4-dihydroxytetrone, reductic acid, and furfural as the five-carbon compound. These products possess an absorption maximum above 240 nm, and most of them are electrochemically active reducing compounds which are derived from nonreducing dehydro-L-ascorbic acid through the processes of delactonization, decarboxylation, and intramolecular rearrangement. PMID- 8250253 TI - A microtiter plate assay for cytochrome c oxidase in permeabilized whole cells. AB - Defects of the mitochondrial respiratory chain often involve dysfunction of cytochrome c oxidase, the terminal component of this chain. We have developed a microassay, suitable for various cell types, which can be carried out in microwell plates. This assay has advantages over the traditional spectrophotometric method as only a small number of cells are used, without the need for mitochondrial fractionation. This makes the assay faster and use of the microtiter plate allows many samples to be screened simultaneously. We demonstrate that lowered levels of cytochrome c oxidase activity can be detected in patients' blood samples, avoiding the necessity for a more invasive tissue biopsy. This cytochrome c oxidase assay is also suitable for use as an intramitochondrial marker for monitoring the effect of mitochondrial translation inhibitors or effectors which modulate mitochondrial biogenesis and could be used for assessing the effect of mitochondrial DNA mutations. PMID- 8250254 TI - Electrospray ionization mass spectrometry on hydrophobic peptides electroeluted from sodium dodecyl sulfate-polyacrylamide gel electrophoresis application to the topology of the sarcoplasmic reticulum Ca2+ ATPase. AB - We describe a method to prepare proteins and peptides in a state suitable for exact determination of molecular mass by electrospray ionization mass spectrometry after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) and electroelution. The utility of the procedure, in conjunction with N terminal sequencing, in defining the C-terminal end of the peptide fragments produced by proteolysis of sarcoplasmic reticulum Ca2+ ATPase with V8 is demonstrated. The application of mass spectrometry aids significantly the use of proteolytic enzymes for topological studies of membrane proteins, and SDS-PAGE is preferable to reverse-phase HPLC for separation of membraneous, hydrophobic peptides and proteins. PMID- 8250255 TI - DNA-DNA subtractive cDNA cloning using oligo(dT)30-Latex and PCR: identification of cellular genes which are overexpressed in senescent human diploid fibroblasts. AB - We previously reported an efficient method for subtractive cDNA cloning using oligo(dT)30-Latex and polymerase chain reaction (PCR) (E. Hara et al., Nucleic Acids Res. 19, 7097-7104, 1991). The subtraction was performed by hybridization between mRNA of cell type B and the cDNA made from mRNA of cell type A using an oligo(dT)30 primer covalently linked to Latex particles in an Eppendorf tube. The mRNA common to both types of cells could be removed by a brief centrifugation. In the present paper, the method was improved by using the sense strand DNA instead of mRNA for hybridization to cDNA covalently linked to the particles to minimize mRNA degradation and by optimizing the hybridization condition. The sense strand DNA was made from cDNA-oligo(dT)30-Latex by asymmetric PCR. Using the improved method, a subtractive cDNA library with longer cDNA inserts was successfully constructed with higher probability than the original method. PMID- 8250256 TI - Substrate-gel electrophoresis for composition and molecular weight of proteinases or proteinaceous proteinase inhibitors. AB - A rapid, sensitive, and generally applicable substrate-sodium dodecyl sulfate polyacrylamide gel electrophoresis method for detection of proteinases or proteinaceous protease inhibitors in biological preparations is described. Electrophoretic separation of proteinases or proteinaceous proteinase inhibitors in the sample using sodium dodecyl sulfate-polyacrylamide gel slabs is followed by immersion of the gel in (1) a protein substrate solution for detection of proteinases or (2) an appropriate proteinase solution, and then in a protein substrate solution for detection of proteinase inhibitors. Some advantages of the reported method over previously described techniques that incorporate substrate into the gel matrix are: (1) the development of bioactive bands, as well as staining and washing, is accomplished more quickly, i.e., 4 to 6 h; (2) the trailing of proteolysis which often appears in copolymerized substrate systems is eliminated; (3) the method is applicable to assay at pH values other than those used for electrophoresis; (4) molecular weight markers can be visualized on the same gels; (5) proteinaceous proteinase inhibitors can also be examined in the biological samples; and (6) the sensitivity is several times higher than that of former assays. PMID- 8250257 TI - A novel ES cell line, TT2, with high germline-differentiating potency. AB - In producing mutant mice by gene-targeting and gene-trapping in embryonic stem (ES) cells, the efficient colonization of the mutant ES cells into germline is still a critical matter. We have established a new line of ES cells, TT2, from an F1 embryo between a C57BL/6 female and a CBA male. When the TT2 cells were injected into blastocysts, the colonization into each tissue was very low. However, when injected into eight-cell embryos, the cells segregated inside the blastomeres, localized in an inner cell mass of blastocysts developed 1 day later, and colonized efficiently in each tissue of the pups. The pups were disproportionately male, about half of which were composed of TT2-derived cells primarily; in more than 70% of the males, TT2-derived cells were dominant, accounting for over half of the total cells. When these males were mated, they exclusively yielded TT2-derived offspring. The germline-differentiating potency was stable during 3 weeks of culture. Twenty-one of 24 mutant clones independently isolated yielded germline chimeras, and 19 clones yielded them in a rate comparable to that of the parent cells. Thus, TT2 cells can serve as a valuable vehicle for the production of mutant mice. PMID- 8250258 TI - A novel negative selection for homologous recombinants using diphtheria toxin A fragment gene. AB - In producing mutant mice by gene targeting in embryonic stem (ES) cells, the efficient isolation of the homologous recombinants is still a critical step. We previously reported on a negative selection using the diphtheria toxin A (DT-A) fragment gene for homologous recombinants (1). It was efficient but limited to gene loci expressed in ES cells. For wider applicability of this negative selection to many gene loci not expressed or expressed at low levels in ES cells, we exploited a novel targeting vector composed of a polyA-less neo gene, a mRNA destabilizing signal, a pausing signal for RNA polymerase II from the minute virus of mice, and the DT-A gene. There was about a 30-fold decrease in frequency of G418-resistant colonies with this strategy against that using only the neo gene in the vector, and homologous recombinants were obtained at frequencies of more than 1/50 among G418 resistant cells at fyn, csk, c-mos, and insulin receptor substrate-1 gene loci. PMID- 8250259 TI - High-sensitivity gas phase sequence analysis of proteins and peptides on PVDF membranes using short cycle times. AB - An optimized sequencer program with a cycle time of 38 min which is specifically tailored for analysis using polyvinylidene difluoride (PVDF) membranes has been developed. The program was developed using a pulsed liquid-phase instrument which was converted to gas-phase acid delivery. Gas-phase acid delivery minimized sample extraction from PVDF membranes and improved tryptophan yields in at least some cases. Other modifications which contributed to reliable high sensitivity sequencer performance included use of a Blott cartridge, substitution of ethyl acetate:heptane (1:1, v/v) instead of butyl chloride as the extraction solvent, use of a modified 100-microliters injection loop with an internal restrictor to reliably inject nearly 90% of the sample, and an HPLC gradient which resolved tryptophan from diphenylurea. These shortened cycle times were achieved at the conventional gas-phase reaction temperature. A slight increase in lag or carryover at prolines was compensated by reduced background from nonspecific acid cleavage which facilitated extended and/or high sensitivity sequencing of large proteins. Reproducible high initial and repetitive cycle yields were obtained with a wide range of experimental peptides which were electroblotted from either 1D or 2D polyacrylamide gels onto high retention PVDF membranes. Initial yields of the majority of the experimental samples analyzed with this program were less than 5 pmol. In addition, most samples with initial yields below 1-2 pmol yielded sufficient sequence information to identify the protein by comparison to protein sequence data-bases or to design oligonucleotide probes. PMID- 8250260 TI - Detection of proteins on polyacrylamide gels using calconcarboxylic acid. AB - We describe here a protein staining method in polyacrylamide gels with a new staining dye, 1-(2-hydroxy-4-sulfo-1-naphthylazo)-2-hydroxy-3-naphthoic acid (calconcarboxylic acid, NN). This method can be performed by both simultaneous and postelectrophoretic staining techniques. Simultaneous staining using 0.01% of NN in upper reservoir buffer eliminates the poststaining step, and thus enables detection of the proteins more rapidly and simply. In poststaining, proteins can be stained by a 30-min incubation of a polyacrylamide gel in 40% methanol/7% acetic acid solution of 0.05% NN. These techniques produced protein staining patterns identical to the ones obtained by the conventional poststaining with Coomassie blue R-250 (CB). NN staining can detect as little as 10 ng of bovine serum albumin by poststaining and 25 ng by simultaneous staining, compared to 50 ng detectable by CB poststaining. In comparing the relationship between band intensity and amount of protein, NN staining gave better linearity than CB staining. PMID- 8250261 TI - Solute adsorption on polymer-coated fused-silica capillary electrophoresis columns using selected protein and peptide standards. AB - Three series of basic proteins and peptides were selected for use in studying the adsorption behavior of fused-silica capillary electrophoresis columns. A comparative study of untreated and treated fused-silica columns was done. The polymers used as coating materials ranged from very hydrophobic to very hydrophilic. The average efficiency of weakly hydrophilic Ucon-coated capillaries for basic proteins was over 1 million theoretical plates at a buffer pH of 4.8. For this coating, it was found that the maximum column efficiency was obtained at a film thickness of approximately 100 nm. It was easily seen that both ionic and hydrophobic interaction forces can play important roles in protein adsorption onto the capillary walls. Furthermore, strongly hydrophilic polymer coatings may possibly lead to hydrogen-bonding interactions, which decrease column efficiency. Studies have been conducted using two series of polypeptides with varying chain lengths. One series was used to study the effect of polypeptide chain length on retention behavior. Efficient separations of bioactive peptide drugs on Ucon coated columns were achieved. The results indicate that high efficiency and resolution of basic peptides tested can be realized on columns modified with weakly hydrophilic polymers at a pH value of about 5. PMID- 8250262 TI - Site-specific carbohydrate identification in recombinant proteins using MALD-TOF MS. AB - The utility of matrix-assisted laser desorption time-of-flight (MALD-TOF) mass spectrometry for the analysis of recombinant glycopeptides is discussed and compared to information which may be obtained by fast atom bombardment mass spectrometry (FABMS) and tandem mass spectrometry (MS/MS). MALD-TOF appears to be 10-100 times more sensitive than FAB MS for the analysis of underivatized glycopeptides, providing qualitative site-specific information regarding the carbohydrate microheterogeneity without the extensive isolation and derivatization procedures required to obtain similar information by FAB MS. Analysis of a digest mixture in the positive and negative ion mode of MALD-TOF indicated that, in mixtures, sialylated glycopeptides are preferentially detected in the negative ion mode. The determination of the molecular masses of a glycopeptide with MALD-TOF prior to and after treatment with a variety of specific glycosidases, often without removal of the buffers, coupled to a comparison of molecular mass information available from a carbohydrate database facilitates the assignment of a carbohydrate composition. The vast majority of the molecular ion signal observed in the linear mode for sialylated glycopeptides are metastable ions. Reflector mass spectra reveal a shift to lower mass consistent with the loss of most of the neuraminic acid residues. The loss of Hex and HexNAc residues is also observed. Sequential lowering of the reflector potential reveals structurally significant fragment ions representing the carbohydrate and peptide portions of the molecule. PMID- 8250263 TI - Differentiation of Ca(2+)- and Mg(2+)-coordinated branched trisaccharide isomers: an electrospray ionization and tandem mass spectrometry study. AB - Three isomeric branched trisaccharides coordinated to calcium and magnesium were analyzed by electrospray ionization followed by tandem mass spectrometry. Studies were undertaken to optimize the singly charged metal coordinated precursor prior to MS/MS and MS/MS/MS experiments. The positioning of the glycosidic linkages between reducing and nonreducing rings could be determined in all three isomers, and the linkage of the fucose ring was obvious in two of the three isomers when calcium was used as the coordinating metal. The ionic radius and charge of the metal appear to be very important in obtaining structural information in these isomers. PMID- 8250264 TI - Optimization of electric field strength for DNA sequencing in capillary gel electrophoresis. AB - Capillary gel electrophoresis (CGE) has demonstrated the ability to separate DNA sequencing reactions at speeds up to 25 times as great as conventional slab gel electrophoresis. These increased speeds are made possible by the efficient heat dissipation of capillaries, which permits higher electric fields to be employed without deleterious thermal effects. The high electric fields, however, also lead to a reduction in the spacing between bands with a concomitant loss of resolution. The resulting tradeoff between speed and resolution is a very important practical aspect of these high-field separations. This work addresses this question by investigating the band broadening and resolution of DNA fragments as they are separated through a fixed distance of gel at field strengths ranging from 50 to 400 V/cm. It is found that the bandwidths of DNA fragments do decrease with the higher field strengths due to a reduction in the diffusional broadening of bands. However, at sufficiently high electric field strengths, the bands begin to broaden again due to the thermal gradient across the gel. This behavior causes the optimum electric field strength for maximum fragment resolution to depend upon the length of fragments being separated. The relative contributions of diffusion and thermal gradients are discussed and used to predict the ultimate performance of constant field capillary gel electrophoresis. PMID- 8250265 TI - DNA sequencing by capillary electrophoresis with replaceable linear polyacrylamide and laser-induced fluorescence detection. AB - Replaceable linear polyacrylamide (LPA) has been utilized as a sieving matrix for DNA sequencing by capillary electrophoresis (CE). Difficulties associated with cross-linked polyacrylamide gel stability have been overcome for the routine application of CE to DNA sequencing. A simple laser-induced fluorescence (LIF) detection system based on a single laser and two photomultipliers (PMT) has been adopted for this work. Sequencing information for four bases has been obtained from two fluorescent dyes and two peak height ratios, detected in two optical channels. FAM- and JOE-labeled M13 (-21) primers have been chosen because both dyes are efficiently excited with a low-power argon ion laser, can be optically separated, and exhibit minimal dye-based shifts in DNA fragment mobilities. Addition of denaturants to the electrophoresis running buffer (1 x TBE, 3.5 M urea, 30% formamide) and column operation at 32 degrees C permitted the resolution of difficult compressed sites in the sequence of phage M13mp18. Careful examination of the polymerization reaction of LPA has led to methodology that has proven to be reproducible for obtaining DNA sequencing information of M13mp18 phage for 350 nucleotides in close to 30 min. PMID- 8250266 TI - Fragmentation of protonated peptides: surface-induced dissociation in conjunction with a quantum mechanical approach. AB - This paper describes the results of a systematic investigation designed to assess the utility of surface-induced dissociation in the structural analysis of small peptides (500-1800u). A number of different peptides, ranging in mass and amino acid sequence, are fragmented by collision with a surface in a tandem mass spectrometer and the spectra are compared with data obtained by gas-phase collisional activation. The surface-induced dissociation spectra provide ample sequence information for the peptides. Side-chain cleavage ions of type w, which are generally detected upon kiloelectronvolt collisions with gaseous targets but not upon electronvolt collisions with gaseous targets, are detected in the ion surface collision experiments. A theoretical approach based on MNDO bond order calculations is suggested for the description of peptide fragmentation. This model, supplemented by ab initio calculations, serves as a complement to the experimental work described in the paper and explains (i) the easy cleavage of the amide bond, (ii) charge-remote backbone and side-chain cleavages, and (iii) the influence of intramolecular H-bonding. PMID- 8250267 TI - Quartz crystal microbalance for the detection of microgram quantities of human serum albumin: relationship between the frequency change and the mass of protein adsorbed. AB - We have developed a piezoelectric immunosensor for the detection of microalbumin. Human serum albumin (HSA) in the range 0.1-100 micrograms mL-1 could be detected using a flow cell; the immunosensor is sensitive enough to monitor levels of albuminuria. The immunosensor did not respond to bovine serum albumin, only to HSA, implying that the specificity for HSA was high. We investigated the relationship between the frequency change (delta F) and adsorption per unit area of piezoelectrically active quartz crystal (delta M). delta M was estimated with radioisotope-labeled anti-HSA or HSA. When anti-HSA was adsorbed onto the surface of the crystal or HSA was bound to anti-HSA supported by the crystal, values of magnitude of delta F/delta M were larger than the value predicted from theory (Sauerbrey's equation). Furthermore, magnitude of delta F/delta M for HSA was larger than that for anti-HSA. PMID- 8250268 TI - A contribution to the history of renal structure knowledge (from Galen to Malpighi). AB - This work documents the progressive gain in knowledge on renal anatomy acquired by anatomists from Galen to Malpighi. Galen, with albeit his rather imaginative explanations, was the first anatomist to recognize the urine producing function of the kidney. His influence was felt up to the Middle Ages; his followers imagined the presence of two cavities within the kidney that were separated by a porous membrane that they called the "colatorium" which was capable of filtering the urine from the blood. It was only later that Berengario da Carpi, divorcing himself from Galenic dogmatism, finally dismissed the presence of the colatorium. He described the renal papillae and gave the first elementary model of renal vascularization. Further important progress was due to the studies of Falloppius and Eustachius who brought contemporary understanding of renal structure to the limit of what could be seen with the unaided eye. They distinguished the difference between the unilobar canine kidney and the human multilobar organ, they also described the minor and major calyces and, furthermore, guessed at the canalicular structure of its parenchyma. Highmore was then responsible for the description of the archiform vessels which he hypothesized as an arterio-venous anastomotic net between the renal cortex and medulla. With the invention of the microscope, new doors opened for the study of renal structure. Bellini proved the canalicular organization of the parenchyma and, moreover, described the interlobular vessels. Malpighi then described the glomerulus and its relation to the intrarenal excretory ducts. The basis had now been laid for the beginning of modern nephrology. PMID- 8250269 TI - The neurons of the human magnocellular septal nuclei: a Golgi study. AB - The morphology of neurons in the magnocellular septal nuclei (medial septal and diagonal band nucleus) were studied in frontal sections of 15 human brains by means of the Golgi method. We classified neurons in the diagonal band nucleus according to their size and morphology into four types: type I--multipolar neurons, type II--fusiform neurons, type III--triangular neurons and type IV- fusiform multipolar neurons. The neurons of the medial septal nucleus we classified into two types: type I--multipolar neurons and type II--fusiform neurons. Our results indicated greater morphological variability of neurons in the human diagonal band nucleus than in the medial septal nucleus. PMID- 8250270 TI - Scanning electron microscopy of the Rhodnius neglectus (Hemiptera) labial salivary glands after starvation. AB - Labial salivary glands are found in the majority of insects. They are relatively large, extend back into the thorax, and in Rhodnius, they are cherry red in color due to a pigment derived from traces of hemoglobin absorbed form the gut. In most insects they are acinous shaped, with long excretion channels that present differentiated regions which from salivary reservoirs. The glands may be relatively simple or complexly branched and convoluted. In Rhodnius they are described as being unilobed with no traces of division. The main duct leaves the gland at its anterior extremity. The acini have different kinds of cells but all of them are seen as sources of secretion. Our material has a different shape due to the fact that the animals spent 20 days under starvation conditions. New data are also obtained through treatment with collagenase and HCl. The importance of the study of these glands lies in the fact that it will further understanding of the transmission of Chagas' disease. PMID- 8250271 TI - Electronmicroscopic observation on the degeneration of skeletal muscles in Xenopus laevis during metamorphosis and after denervation. AB - We observed metamorphosis-related structural changes in the tail muscle of Xenopus laevis tadpoles during tail involution and changes in the calf muscles of adult Xenopus laevis. The specimens were treated with hydrochloric acid for extraction of the fibrous components in the connective tissues. Prior to metamorphosis, the muscle fibers appeared straight and flat. At the start of metamorphosis, they appeared crooked with uneven surfaces. Scanning electronmicroscopy revealed a marked increase in the number of macrophages and fibroblast-like cells in the muscle fibers. Transmission electronmicroscopy revealed an enlargement of the sarcoplasmic reticulum, the appearance of myelin structures and the uptake of muscle fragments by macrophages. The myofilaments in the muscle fragments were in different stages of decomposition. After denervation of the nervus ischiadicus, scanning electron-microscopy showed that the diameter of the muscles was reduced; a fine unevenness of the muscle surface could be observed. However, the cellular component in the muscle fibers was not obviously increased. Transmission electronmicroscopy showed that the sarcolemma and basal lamina were finely waved; cross striae were unclear due to a decrease in myofilaments. There are similarities in the structural changes which occur in the tail muscles of Xenopus laevis during tail involution and calf muscles after denervation, but the processes are not identical. In tadpols, nerves exert a secondary influence on structural changes of the muscles in the process of tail involution during metamorphosis. PMID- 8250272 TI - Morphological studies on the rodent main and accessory olfactory systems: the regio olfactoria and vomeronasal organ. AB - The present study on the main olfactory system (MOS) and the accessory olfactory system (AOS) documents the functional morphology of the rodent olfactory region and that of the vomeronasal organ (VNO) using light and electron microscopical techniques. Special attention is given to the cytoarchitecture of the sensory epithelia, i. e. the olfactory epithelium (OE) of the regio olfactoria and the neuroepithelium of the VNO (VNO-NE). Both sensory epithelia consist of a pseudostratified columnar epithelium composed of three types of cells, i. e. receptor cells, supporting cells and progenitor cells. Even at the light microscopical level, however, distinctive morphological features can be distinguished which illustrate important differences between the two sensory epithelia. For example, the height of the respective epithelia differs considerably, the VNO-NE is approximately 170 microns tall and the OE is only about 90 microns. The receptors of the VNO-NE lack olfactory knobs which are typically found in the sensory cells of the OE. The perikarya of the receptor cells of the VNO-NE are very large when compared to those of the sensory cells of the OE. In contrast to the OE, blood vessels are found within the neuroepithelial layer of the VNO. The progenitor cells of the OE are located in a clearly distinguishable cell layer which is lacking in the rodent VNO-NE. The differences between the two epithelial layers become more obvious at the electron microscopical level. The olfactory knobs of the sensory cell dendrites of the OE reach the nasal cavity with numerous cilia. These olfactory hairs, on average 11 per knob, consist of a short proximal segment and a long and thin distal segment. This distal segment runs parallel to the epithelial surface and is embedded in the neuroepithelial mucosal layer. The dendrites of the receptor cells of the VNO NE reach the lumen of the VNO with numerous branched microvilli which are also embedded in the mucous layer. Horizontal ultrathin sections through the apical portion of the OE reveal that each supporting cell completely envelopes several dendrites. This glia-like relationship is not found in the corresponding layer of the VNO-NE. The sensory cell perikarya of the OE contain only a few endoplasmatic reticulum (ER) profiles while the receptor cells of the VNO are characterized by an extensive smooth endoplasmatic reticulum (SER). In contrast to the fila olfactoria, numerous axons within the vomeronasal nerve show ellipsoidal varicosities without synaptic vesicles which may indicate the existence of at least two vomeronasal nerve fibers.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8250273 TI - Ultrastructural study of the testis of non-breeding naked mole-rat (Heterocephalus glaber, Ruppell). AB - The testicular structure of the wild caught naked mole rat was studied. It comprises of a large volume of lipid-rich interstitial cells of Leydig among which are few scattered seminiferous tubules. In addition, the interstitial cells possess elongated mitochondria and vast network of smooth endoplasmic reticulum (sER). The Golgi apparatus (GA) apparently is not conspicuous or well developed. All stages of spermatogenesis occur in the seminiferous tubules although the mature forms (secondary spermatocytes, spermatids and spermatozoa) are few. Sertoli cells show an irregular nucleus, mitochondria oriented perpendicular to the basement membrane, a vast network of endoplasmic reticulum with sER as the predominant form and lipid droplets. The ultrastructural features of Leydig cells seem to suggest a steroidogenic capacity although the vast accumulation of lipid droplets may imply impaired utilisation of cholesterol reservoir as a result of pituitary hormonal imbalance or (and) the local paracrine influence by Sertoli cells. The cause of slow-down in spermatogenesis is still unclear but may also be under the influence of pheromonal cues or the local paracrine control. Sertoli cell features point towards a role of synthesis and secretion. PMID- 8250274 TI - Morphometric evaluation of the rat testis, epididymis and vas deferens following chemical sympathectomy with guanethidine. AB - Selective chemical sympathectomy of the internal sex organs of adult male rats was undertaken by long term administration of low doses of guanethidine. The spermatogenic activity of the testis was unaffected by treatment. Examination of the vas deferens using morphometric methods revealed a marked increase in luminal area in contrast to a decrease in muscle layer area and in epithelial height. This is morphological evidence of sperm accumulation caused by a disorder in ductal contractile activity. No structural changes were observed in the epididymis. However, the concentration of spermatozoa in the sperm suspension stored in the cauda epididymidis was significantly increased in denervated rats. This result is discussed in terms of a sympathetic control of resorption mechanisms in the epididymis. PMID- 8250275 TI - [Prenatal development of the radial annular ligament]. AB - The prenatal development of the annular ligament of the elbow was studied by investigating 600 microns thick plastinated sections through the elbow joints of nine fetuses with a crown-rump-length (CRL) of 105-360 mm. At 105 mm CRL the annular ligament consists of one layer of circular fibres around the circumference of the radial head. At 130 mm CRL it extends further distally to the radial neck. The postero-lateral region exhibits an arrangement of three layers of fibres of different course. Stages later than the 190 mm stage show in the anterior region external fibres arising from the medial periosteum of the ulna. They pass over the anterior surface of the ulna and blend with fibres arising from the anterior margin of the ulna's radial notch to surround the radial head. At 270 mm the articular capsule can be recognized as a strong homogenous layer of fibres. With all stages investigated the annular ligament consists of circular fibres only near the attachment at the posterior margin of the ulna's radial notch. Anteriorly the annular ligament is narrower than posteriorly. With the later stages of 290, 330 and 360 mm CRL a further differentiation of the annular ligament cannot be recognized. Thus, between 105 mm and 270 mm CRL, the annular ligament develops a more and more complex structure similar to that described for the adult joint. We describe several distinct steps of this differentiation during the fetal period which are not mentioned in the literature yet. PMID- 8250276 TI - Monochromatic x-ray radiation for high resolution radiography in osteology. AB - The contrast of an x-ray exposure is influenced by the x-ray absorption of the examined material and the x-ray frequency. The examination of the absorbing properties of bone material in the molybdenum spectrum by means of the Bragg method revealed an intense increase of absorption in the long-wave range. By using a zirconium filter, long-wave and monochromatic MoK alpha-rays can be isolated from the spectrum of a molybdenum x-ray tube. The x-ray exposures of thin bone tissue produced in this spectral range show a much greater degree of contrast and depth as compared to conventional exposures. PMID- 8250277 TI - Growth allometry of the human face: analysis of the osseous component of the mid and lower face in Brazilian fetuses. AB - Growth of facial bones was studied in 30 human fetuses ranging from 14 to 35 weeks of gestation (both sexes together), investigating both bivariate and multivariate allometry with principal components analysis (PCA). Weight of the mandible, maxillae, zygomatic, palatine, vomer and both lateral masses of the ethmoid bones was measured and correlated with fetal weight. The relative order of growth rates was similar for bivariate and multivariate results. These results indicate that the growth rate of the lower face is greater than growth rates of the vomer and palatine bones, and is less than growth rates of the maxillae, zygomatic and ethmoid bones. Bivariate and multivariate results present only one significant difference: the mandible grows with positive allometry in bivariate study and with negative allometry in multivariate analysis. The analysis performed in this study provides an approximation of the biological phenomenon of growth which may be useful in the interpretation of facial growth. PMID- 8250278 TI - Epidermal lipid in several cetacean species: ultrastructural observations. AB - The ultrastructure of the skin of four cetacean species, bottlenose dolphin (Tursiops truncatus) long-finned pilot whale (Globicephala melaena), humpback whale (Megaptera novaeangliae), and fin whale (Balaenoptera physalus) was investigated with particular reference to epidermal lipid. It has already been established that massive lipid reservoirs exist in whales, that the biochemical structures of cetacean lipids are unique, and that unusual intracellular lipid droplets appear in the epidermis. We report here some novel findings on scanning electron microscopic morphology of epidermal lipid, and on its ultrastructural morphology in general and specialized integumentary sites, including species not previously investigated. The intracellular epidermal lipid droplets were more extensive than lamellar body-derived intercellular lipid which is within the interstices of stratum externum cells. The intracellular droplets were spherical, highly variable in size ranging from 0.24 micron to 3.0 microns in diameter, appeared singly or were aggregated in cytoplasmic cavitations, and often were closely associated with epidermal cell nuclei. Evidence for exocytosis of the intracellular droplets was not observed. Significant numbers of intracellular lipid droplets are not observed in the epidermis of terrestrial mammals, so their presence is one of several aquatic specializations of the cetacean integument. Its full significance remains obscure, but it is more probably associated with epidermal cell metabolism than with secretion of lipid. PMID- 8250279 TI - The ventralizing effect of the notochord on somite differentiation in chick embryos. AB - The dorso-ventral pattern formation of the somites becomes manifest by the formation of the epithelially organized dorsal dermomyotome and the mesenchymal ventrally situated sclerotome. While the dermomyotome gives rise to dermis and muscle, the sclerotome differentiates into cartilage and bone of the axial skeleton. The onset of muscle differentiation can be visualized by immunohistochemistry for proteins associated with muscle contractility, e.g. desmin. The sclerotome cells and the epithelial ventral half of the somite express Pax-1, a member of a gene family with a sequence similarity to Drosophila paired-box-containing genes. In the present study, changes of Pax-1 expression were studied after grafting an additional notochord into the paraxial mesoderm region. The influence of the notochord and the floor-plate on dermomyotome formation and myotome differentiation has also been investigated. The notochord is found to exert a ventralizing effect on the establishment of the dorso-ventral pattern in the somites. Notochord grafts lead to a suppression of the formation and differentiation of the dorsal somitic derivatives. Simultaneously, a widening of the Pax-1-expressing domain in the sclerotome can be observed. In contrast, grafted roof-plate and aorta do not interfere with dorso-ventral patterning of the somitic derivatives. PMID- 8250280 TI - Immunocytochemical demonstration of nerve growth factor receptor (NGF-R) in developing human fetal teeth. AB - Evidence is accumulating that nerve growth factor receptor (NGF-R or p75NGFR) can mediate cell growth and differentiation of non-neuronal cells. NGF-R expression was studied in developing teeth of human embryos and fetuses between the 6th and 18th weeks of gestation, using a monoclonal mouse-anti-human NGF-R antibody. In contrast to earlier findings in rodents, the NGF-R expression of the human dental papilla was found to be transient. NGF-R was present in the condensing ecto mesenchymal cells of the dental papilla in the early cap stage tooth germ. In later developmental stages, a shift of the NGF-R expression from the papilla to the cytoplasmic membrane of the inner enamel epithelium (IEE) was demonstrated. As in rodent odontogenesis, the NGF-R immunoreactivity of the IEE remained until the odontoblasts started secretion of predentinal matrix in the late bell state. The mitotic activity in the IEE was detected by an antibody against proliferating cell nuclear antigen (PCNA) and showed that the NGF-R expression of the IEE decreased as the cell proliferation ceased. We propose that NGF-R may, be involved in differential and/or proliferative events of human odontogenesis. PMID- 8250281 TI - Microvasculature of regular and irregular areolae of the areola-gland subunit of the porcine placenta: structural and functional aspects. AB - To examine the vasculature of the areola-gland subunit of advanced pig placenta, tissues from ten animals between 43 and 112 days of gestation were prepared for histology and for scanning electron microscopy of vascular corrosion casts from both maternal and fetal sides. Regular areolae, tributary to one gland only, are round with a wide-meshed and smooth subepithelial capillary network on the maternal side, which is similar to the pre-implantative stage and bordered by an abrupt rim towards the inter-areolar maternal capillary network. On the fetal side, the capillary network follows papillae which protrude into the areolar cavity or converge to form a ring towards the areolar periphery. Irregular areolae, in contrast, have indistinct boundaries and are characterized by two or more gland openings. The maternal capillary network has moderate density and follows the corrugations, whereas the fetal capillary network is basically two dimensional with some blunt sinusoidal protrusions. Vessel architecture of both areolar types implies facilitated external inflow of blood into the areola on arteriolar as well as on capillary levels, whereas the outflow from the areolar capillaries comprises venules converging into one or two areolar stem veins, and therefore conducts venous blood in a manner different from that of the inter areolar region. It is suggested that this arrangement could favour vascular control mechanisms in uterus, placenta and fetus. On the basis of these observations and the discussion, it is suggested that these areola-specific vessel systems are important for sustaining the characteristic substance transfers in the areola, the secretion, metabolism and absorption, which according to the literature are not the same in the regular as in the irregular areolar type of the porcine areola-gland subunit of the placenta. PMID- 8250282 TI - Correlation between the embryonic head flexures and cardiac development. An experimental study in chick embryos. AB - The aim of the present study is to examine whether the formation of the cranial and cervical flexures is involved in the process of cardiac looping, and whether looping anomalies are causally involved in the development of cardiac malformations. For this purpose, the formation of the cranial and cervical flexures was experimentally suppressed in chick embryos by introducing a straight human hair into the neural tube. In the experimental embryos, the absence of the cervical flexure, alone or in combination with a reduced cranial flexure, was always associated with anomalies in the looping of the tubular heart. The convergence of the primary distant venous and arterial ends of the heart, as well as the normal movement of the ventricular region from its original position, cranial and ventral from the cardiac inflow, to its final position caudal to the presumptive atria, was suppressed to an extent related to the degree to which the formation of the flexures was prevented. Positional immaturity of the heart loop (increased distance between its inflow and outflow, and cranio-ventral position of the ventricular region) was associated with incomplete deformations (reduced angulations) of the cardiac wall at the atrioventricular or conoventricular junctional areas. Reduced angulations were associated with the hypoplasia of the anlagen of the cardiac septa at the level of the angulation (av-cushions, conal ridges). Hypoplasia of these anlagen was followed by incomplete or absent fusion of their opposite free edges, which finally resulted in atrioventricular or ventricular septal defects. These results show that the convergence of the venous and arterial ends of the tubular heart and the caudo-dorsal movement of its ventricular region are related to the formation of the cervical flexure, and that the mesenchymal septa of the heart seem to develop in response to deformations of the embryonic heart, which are generated by the process of cardiac looping. Therefore, the positional and morphological changes of the looping heart are regarded as playing a key role in the process of normal and abnormal morphogenesis of the heart. PMID- 8250283 TI - An analysis of HRP-filled basket cell axons in the cat's cerebellum. I. Morphometry and configuration. AB - Basket cell axons terminate directly on the Purkinje cell body in the mammalian cerebellum and thus are in a position to have a direct influence on this neuron. In the present study, individual basket cells were intracellularly filled with horseradish peroxidase and the morphological characteristics and distribution of their axons were analyzed and quantified. The horizontally directed axon of basket cells preferentially distributes toward one side of the cell body, and is usually directed toward the base of the folium. In their course, the axons give rise to three types of axonal specializations including descending collaterals that form the complex endings around the axon hillocks and somata of Purkinje cells (i.e. the pinceau), beaded tendrils that arise from the descending collaterals and distribute to the Purkinje cell and upper granule cell layer, and beaded tendrils that arise directly from the horizontal axon to enter the molecular layer. In some respects, basket cells comprise a heterogeneous population of neurons on the basis of the length of their axon, as well as the number, distribution, and morphological characteristics of the axonal collaterals. The major finding of this study is that a single basket cell does not form a pinceau around the axon hillock of every Purkinje cell located along the course of its horizontal axon. Further, the pinceaux that are formed vary in their complexity. These findings suggest that the classic concept of off-beam inhibition mediated by basket cells is more complex than previously described. PMID- 8250284 TI - An analysis of HRP-filled basket cell axons in the cat's cerebellum. II. Axonal distribution. AB - The axons of basket cells course horizontally within the lower molecular layer of the cerebellar cortex and give rise to three types of axon collaterals. One collateral distributes around the somata and axon hillocks of Purkinje cells, forming a complex pericellular basket-like formation or "pinceau". The other two axonal specializations have the form of beaded tendrils, but differ in their origin and laminar distribution. In the previous paper, the morphometry and configuration of these axonal branches were described. In the present account, their synaptic relationships are analyzed based on serial section analysis and computer reconstructions. Our data indicate that: (1) Purkinje cells receive descending collaterals from more than one basket cell; (2) the contribution of the descending collaterals to individual pinceau is not uniform; Purkinje cells located close to the basket cell of origin receive a greater number of descending collaterals when compared to Purkinje cells located more distally; (3) few synaptic junctions are formed by the descending collaterals within the pinceaux; (4) beaded tendrils make synaptic contacts with the somata and dendrites of Purkinje cells, and (5) the horizontal axon forms synaptic junctions with the dendritic shafts and spiny branchlets of Purkinje cells. Functional considerations of these synaptic relationships are discussed in the light of the classic concept of off-beam inhibition mediated by basket cells. PMID- 8250285 TI - Surgical ligation vs. angiographic embolization of the vena spermatica: a prospective randomized study for the treatment of varicocele-related infertility. AB - Varicoceles as a common cause of male infertility are treated either by surgical ligation or, more recently, by angiographic occlusion of the spermatic vein. In the present prospective randomized study 38 patients were treated by surgical ligation and 33 by angiographic embolization. During the 12-month follow-up period a significant increase in sperm number (at 3 and 12 months) and sperm motility (at 12 months) occurred in both groups while sperm morphology remained unaffected. Altogether, 22 pregnancies (31%) were reported within the year following treatment, of which 11/38 (29%) occurred in the ligation group and 11/33 (33%) in the embolization group. Thus both treatment modalities appear equivalent, whereby embolization has the advantage that it can be performed on an outpatient basis. PMID- 8250286 TI - Flow cytometric DNA analysis demonstrates contralateral testicular deterioration in experimental unilateral testicular torsion of prepubertal rats. AB - It has been postulated that unilateral testicular torsion causes damage to the contralateral testis and reduces fertility. However, in animal studies such an effect has not been fully proven by histopathologic examination or other conventional assays of spermatogenesis. We investigated the effect of unilateral testicular torsion on contralateral spermatogenesis in prepubertal rats using quantitative flow cytometric DNA analysis. Male rats were divided into three groups which underwent sham-operation, simple hemiorchiectomy or unilateral testicular torsion. Five weeks after these operations, fertility and spermatogenesis by flow cytometry were evaluated. No significant differences were observed in body weight, contralateral testicular weight or serum testosterone concentration among the three experimental groups. In the torsion group, mean seminiferous tubular diameter, number of foetuses, fertility rate and percentage of haploid cells were all significantly decreased compared to the other two groups. These results suggest that unilateral testicular torsion causes damage to the contralateral testis and consequently can reduce the future fertility of prepubertal rats. PMID- 8250287 TI - Repeated interruptions of the testicular blood flow do not have long-term effects on spermatogenesis in the ram. AB - The effect of repeated interruptions of the testicular blood flow on spermatogenesis was studied in mature Texel rams. Reversible interruption of the blood flow was achieved by an inflatable occluder, placed around the testicular artery at the level of the spermatic cord. In eight testes the blood flow was successfully interrupted six times for 1 h within 3 weeks and in 14 testes nine times for 1 h within 3 weeks. Nine weeks after the last blood flow interruption spermatogenesis was evaluated in histological sections of the testes. Both after six and nine blood flow interruptions a qualitatively complete epithelium was found in at least 90% of the seminiferous tubules. Cell counts in stages VII and VIII of the spermatogenic cycle revealed a slight decrease of spermatocytes and spermatids in the tubules with a complete epithelium after nine occlusions, which was only statistically significant for Preleptotene Spermatocytes. After six occlusions the numbers of all cell types were at or even slightly above control levels. These results show that repeated periods of ischaemia for 1 h do not result in conspicuous long-term damage to spermatogenesis. PMID- 8250288 TI - Protection of spermatogenesis against cytotoxic effects of two chemotherapeutic drugs by temporary testicular blood flow interruption in the ram. AB - Temporary interruption of the testicular blood flow for 1 h after injection of cytostatic drugs has a protective effect on spermatogenesis. This was shown in experiments in which spermatogenesis was evaluated at four weeks after a single intravenous injection of Adriblastina (ADR; doxorubicin hydrochloride) or Mitomycin-C-kyowa (MIT). Interruption of the blood flow was performed by inflation of an occluder implanted around the testicular artery. The animals were killed and histological sections prepared 4 weeks after treatment. In all drug treated animals spermatids were near absence and spermatocytes were decreased in number. Therefore, even after occlusion of the blood flow, the drug doses were high enough to kill not only large numbers of differentiating spermatogonia but also stem cells. The response of the stem cells to the treatments was evaluated by counting the numbers of A spermatogonia per 100 Sertoli cells in the different groups. Normal numbers of these cells were found after both MIT and ADR, indicating that the stem cell population had responded to the initial cell loss by extra proliferation. However, significantly higher numbers of A spermatogonia were found in the drug-treated animals in which the testicular blood flow was interrupted for 1 h. This indicates that occlusion of the blood flow to the testis for 1 h results in a faster recovery of spermatogenesis than after drug treatment alone. PMID- 8250289 TI - Temporal coincidence of the appearance of elongated spermatids and of histochemical reaction of 11 beta-hydroxysteroid dehydrogenase in rat Leydig cells. AB - The serial semithin sections of the testes of perfused rats (age: 25-41 postnatal days) were evaluated microscopically on each alternative day for the morphology of spermatogenetic cells and especially for spermatid differentiation. The histochemical reaction for non-specific esterases, 3 alpha-hydroxysteroid dehydrogenase (HSDH), 3 beta-HSDH, 11 beta-HSDH, 17 beta-HSDH were performed in cryostat sections of corresponding rat testes (age: 25-55 days). Eight different types of germ cell associations (designated as A-H) appeared from day 25 to day 41. Seven steps of differentiation of spermatids were characterized. The turning point of spermatid maturation was observed on day 35. Prior to this day, mainly spermatids step 1 were identified. On day 35 spermatids steps 1-5 were present. The reactions of non-specific esterases, 3 beta-HSDH, 17 beta-HSDH and 3 alpha HSDH were observed in the Leydig cells of all testes. Only from day 35 onwards could 11 beta-HSDH be demonstrated. The temporal sequence of the differentiation of spermatids and the appearance of histochemical activity of 11 beta-HSDH in the Leydig cells seem to be closely correlated. An additional observation concerning the presence of germ cell debris was made in the seminiferous tubules during the postnatal development. Large amounts of debris were observed only till day 33. PMID- 8250290 TI - Purification and characterization of sperm-coating antigen, identified by a monoclonal antibody. AB - A procedure was designed for purification of a human seminal plasma-specific antigen (HSP-antigen) identified by a monoclonal antibody produced in this laboratory (mAb 4E6). Pooled human seminal plasma was fractionated by consecutively applied methods: affinity chromatography on Lentil lectin sepharose, gel chromatography on Ultrogel AcA 34 and immunoaffinity on mAb 4E6 coupled CNBr-Sepharose 4B. The antigen-containing fraction obtained after this procedure was proved to be homogeneous when analysed by electrophoresis in polyacrylamide gel. After the consecutive purification procedures the degree of purification was 128 times as compared to the starting material. Electrophoretic analysis of the purified 4E6 antigen under reducing conditions showed that it consisted of 3 polypeptide subunits with molecular weight 70 kDa, 64 kDa and 60 kDa respectively. On the basis of the data obtained from competitive ELISA testing of sera from infertile patients it has been suggested that the identified antigen may be involved in pathogenesis of immunologic infertility. PMID- 8250291 TI - Binding of a specific subclass of immunoglobulins by a human seminal plasma component. AB - Human seminal plasma contains a component that binds immunoglobulins (Ig). The Ig binding factor was purified by ammonium sulphate precipitation, preparative isoelectrofocusing and gel filtration chromatography and found to bind strongly human IgGl and mouse IgM. This seminal plasma component may possess immunosuppressive activity and may modulate the activities of the immunosurveillance system of the reproductive tract. PMID- 8250292 TI - A hCG-secreting testicular seminoma revealed by male infertility: mechanism of hCG-evoked endocrine disturbances. AB - We report a case of a hCG-producing testicular seminoma revealed by a male infertility due to oligozoospermia. No palpable tumour was found at clinical examination and the diagnosis was established on hormonal data and the echography of the testis. The endocrine patterns associated high levels of beta-hCG and 17 beta-oestradiol contrasting with low levels of LH and FSH and normal levels of testosterone. Immunohistochemical studies confirmed the ectopic production of hCG. Histological findings and the evolution of hormonal parameters suggested that hyperoestradiolaemia was probably the consequence of a hCG-evoked Leydig cells hyperplasia involving a paracrine mechanism. In fact, after removal of the tumour, a dramatic decrease of plasma sex steroid levels was observed before recovery of normal testicular endocrine and exocrine functions. PMID- 8250293 TI - Effects of cyproterone acetate on the reproductive tract and pituitary-gonadal axis of the golden hamster. AB - The effects of cyproterone acetate (CA) on the reproductive tract and serum levels of gonadotropins and testosterone in male golden hamsters exposed to long days, were studied. The daily injection of CA at 100 mg kg-1 body weight during 8 weeks reduced sperm count, both in caput and cauda epididymidis. The weight of epididymis and seminal vesicles were also reduced, while testicular weight and histology were not affected. Serum levels of testosterone and LH were increased 3.6 and 1.8 fold, respectively, above controls, while FSH was not affected by the treatment with CA. The absence of an inhibitory action of CA on the testis could be explained through an increase in LH stimulation, thus suggesting that in the golden hamster CA is devoid of progestagenic partial effects. PMID- 8250294 TI - Expression of the proliferation-associated Ki-67 antigen in bovine testicular tubular cells during the seminiferous epithelial cycle, demonstrated with the MIB 1 antibody. AB - Ki-67 expression in the seminiferous tubule of the bovine testis was studied by immunohistochemistry during the seminiferous epithelial cycle using the monoclonal antibody MIB-1. Spermatogonial proliferation is most obvious in stages 5-7, and 8, when B-spermatogonia divide. A lower rate of spermatogonial propagation is observed preceding or during meiosis in stages 1-4. The MIB-1 antibody also gives positive results with some post-spermatogonial tubular cells. Preleptotenes passing through S-phase in stage 1 reveal positive nuclei. During prophase of meiosis I pachytenes react strongly, diplotenes react in an attenuated manner, while leptotenes and zygotenes stay negative. Secondary spermatocytes seen in stage 4 are positive as are the chromosomes during meta- and anaphase of the meiotic divisions. Post-meiotic spermatids are also decorated but stop Ki-67 expression abruptly at the end of stage 4. Sertoli cells are negative. PMID- 8250295 TI - The role of transesophageal echocardiography in patients with cardiac trauma. PMID- 8250296 TI - The effect of halothane on the sarcoplasmic reticulum in normal and right ventricular hypertrophy in rabbits. AB - Calcium accumulation and storage by the sarcoplasmic reticulum is affected adversely by both inhaled anesthetics and hypertrophy. The interaction of these two conditions in right ventricular muscle was examined in a skinned fiber preparation whereby the sarcolemma was disrupted mechanically to permit external control of the chemical composition of the cytoplasm. Skinned fibers were obtained from the right ventricles of rabbits 4 mo after surgical banding of the pulmonary artery (n = 5) or sham banding (n = 4). Right ventricles after pulmonary artery banding weighed an average of twice that of normal right ventricles. The sarcoplasmic reticulum of the skinned fibers was permitted to accumulate Ca2+ and then maximal or submaximal Ca2+ release was induced with 25 mM or 2 mM caffeine, respectively. The magnitude of the Ca2+ release was estimated by the area of the ensuing tension over time. When 1%, 2%, or 3% halothane accompanied the 2 mM caffeine, both normal and hypertrophied muscle exhibited an area of the tension transient twice that of the tension transients without halothane (control). When present during Ca2+ accumulation, 1%, 2%, and 3% halothane reduced the 25 mM caffeine-induced tension transient in normal and hypertrophied muscle to 36%, 17%, and 11%, respectively, of the controls. The equal depression in normal and hypertrophied right ventricles was in contrast to a previous study in which hypertrophy provided some protection against the decrease in sarcoplasmic reticulum Ca2+ storage by halothane.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250297 TI - Protamine and left ventricular function: a transesophageal echocardiography study. AB - The effect of protamine sulfate on left ventricular function was evaluated in 23 patients undergoing heart surgery requiring cardiopulmonary bypass (CPB). Ventricular function was assessed by measuring cardiac index with a thermodilution pulmonary artery catheter and fractional shortening with transesophageal echocardiography (TEE). After CPB, a heparin-neutralizing dose of protamine was infused continuously for 5 min. Hemodynamic variables were obtained and fractional shortening was measured before protamine administration (time 0), during protamine infusion (2.5 and 5 min), and after the infusion (10 and 15 min after the start of protamine). Patients with a preoperative ejection fraction > or = 50% had no statistically significant changes in either cardiac index or fractional shortening with protamine administration, when compared to time 0. However, patients with a preoperative ejection fraction < 50% had a significant decrease in fractional shortening which occurred during and immediately after protamine administration when compared to time 0 (P < 0.01), with no associated change in hemodynamics. Our results suggest that protamine may have a negative inotropic effect that is apparent in patients with impaired ventricular function. PMID- 8250298 TI - Relation between oxygen consumption and oxygen delivery in patients after cardiac surgery. AB - The relative contributions of oxygen delivery (DO2) and oxygen extraction (O2ER) to the increase in cellular oxygen uptake (VO2) after cardiopulmonary bypass were studied prospectively in 36 patients after coronary artery bypass grafting (n = 18), valve replacement (n = 17), and removal of a left atrial tumor (n = 1). VO2 was calculated from the Fick equation and DO2 from thermodilution cardiac output and arterial oxygen content. During the first 24 h after cardiac surgery, there was a strong relation between VO2 and DO2 (VO2 = 28 + 0.27 x DO2, r = 0.79, P < 0.0001) but not between VO2 and oxygen extraction. Mixed venous oxygen saturation (SVO2) was usually reduced when cardiac index was below 2.0 L.min-1.m-2. Patients with a prolonged intensive care unit course (> 24 h) had lower cardiac index and lower SVO2 than the other patients. Therefore, the progressive increase in VO2 after cardiac surgery is accomplished primarily by an increase in cardiac output and DO2. It is usually when cardiac function is compromised that O2ER increases and SVO2 decreases. PMID- 8250299 TI - Accuracy of continuous jugular bulb venous oximetry during cardiopulmonary bypass. AB - We evaluated the accuracy of fiberoptic catheter oximetry in the jugular bulb during conditions of normothermia, hemodilution, and hypothermia in 11 patients who underwent cardiac surgery with cardiopulmonary bypass (CPB). An oximetry catheter was inserted into the right jugular bulb under general anesthesia, calibrated by the in vitro (n = 7) or in vivo (n = 4) mode. Jugular bulb oxygen saturation (SjO2) with the catheter oximeter was compared with a concurrent laboratory CO-oximeter value from a blood sample during surgery. Nasopharyngeal temperature (NPT) and hemoglobin concentration (Hb) were also measured. The oximetric catheter SjO2 correlated closely with the CO-oximeter determinations in both calibration modes (in vitro, r2 = 0.88; in vivo; r2 = 0.96). Data in the in vitro calibrated group were grouped into three conditions; 1) normothermia and no hemodilution, 2) normothermia and hemodilution, and 3) hypothermia and hemodilution, and showed good correlations between SjO2 values measured by the two methods (r2 = 0.90, r2 = 0.81, r2 = 0.79, respectively). The difference in SjO2 values by the two methods was not significantly affected by changes in NPT and Hb during CPB. In conclusion, the continuous SjO2 monitoring with catheter oximetry during CPB would be accurate and reliable under either calibration mode. Moderate hypothermia and hemodilution during CPB did not significantly influence the accuracy. PMID- 8250300 TI - Positive end-expiratory pressure ventilation decreases plasma atrial and brain natriuretic peptide levels in humans. AB - To investigate the possible role of atrial and brain natriuretic peptides (ANP and BNP) in the renal effects of mechanical ventilation with positive end expiratory pressure (PEEP), we measured changes in plasma ANP and BNP levels during PEEP in patients undergoing subtotal esophagectomy. Application of 15 cm of H2O PEEP for 1 h decreased the levels of plasma ANP and BNP from 24.4 +/- 5.5 (mean +/- SEM) and 19.0 +/- 3.5 fmol/mL to 14.4 +/- 2.1 and 15.3 +/- 3.0 fmol/mL, respectively (P < 0.05). The level of plasma cyclic guanosine monophosphate, an intracellular second messenger of ANP and BNP, also decreased from 8.4 +/- 1.5 to 5.7 +/- 0.8 pmol/mL (P < 0.05). PEEP increased the levels of plasma arginine vasopressin from 2.0 +/- 0.5 to 4.2 +/- 1.2 pg/mL, aldosterone from 36.1 +/- 4.9 to 65.3 +/- 12.7 pg/mL, and plasma renin activity from 1.4 +/- 0.5 to 2.7 +/- 0.7 ng.mL-1.h-1. During PEEP ventilation, urine output, urinary sodium and potassium excretion, osmolar clearance, and cardiac index all decreased. PEEP increased free water clearance, right atrial pressure, pulmonary arterial pressure, and pulmonary capillary wedge pressure. The level of plasma endothelin, mean blood pressure, and heart rate did not change significantly. These results suggest that not only hemodynamics and the vasopressin and renin-angiotensin-aldosterone system, but also the natriuretic peptide system (ANP and BNP), are involved in the renal effects of PEEP. PMID- 8250301 TI - Alfentanil attenuates the cardiopulmonary response of critically ill patients to an acute increase in oxygen demand induced by chest physiotherapy. AB - Critically ill patients often are subjected to interventions that acutely increase oxygen demand and require increased output of the cardiac and respiratory systems. This study explored whether alfentanil could attenuate the response to chest physical therapy, a procedure that increases oxygen consumption by 40%-50%. Patients were examined during two consecutive therapy sessions. In random order, they received either a placebo or alfentanil (30 or 60 micrograms/kg) 2 min before treatment. In Group 1 (n = 11, 30 micrograms/kg alfentanil) only the arterial blood pressure increases induced by chest physical therapy were attenuated. In Group 2 (n = 12, 60 micrograms/kg) alfentanil attenuated the increases in heart rate, central venous pressure, and pulmonary artery systolic pressures as well as systemic blood pressure. Neither dose of alfentanil altered the increases in oxygen consumption, carbon dioxide elimination, oxygen delivery, or extraction ratio. Thus alfentanil attenuated the hemodynamic responses to chest physiotherapy in a dose-dependent fashion. This was likely due to its vagotonic actions. In contrast, alfentanil had no effect on the balance between oxygen demand and delivery during chest physiotherapy. There was thus a dissociation between the hemodynamic and metabolic responses. PMID- 8250302 TI - Pulmonary capillary pressure measured with a pulmonary arterial double port catheter in surgical patients. AB - We developed a pulmonary artery (PA) double port catheter technique for reliable clinical measurements of pulmonary capillary pressure (Ppc). In seven elective surgical patients, the PA double port catheter with the second PA port 1 cm proximal to the balloon was inserted. The two PA ports, connected to identical pressure measuring systems, provided the pulmonary arterial pressures (Ppa) distal and proximal to the balloon. After general anesthesia was stabilized, the two Ppas were measured simultaneously during a PA occlusion maneuver during 10 s of apnea. The instant of occlusion was determined precisely as the time when the two Ppa traces sharply diverged. A single exponential equation was fitted to the segment of distal Ppa tracing starting 0.3 s after the instant of occlusion. Ppc was determined as the value of the exponential fit extrapolated to time 0. In six of seven patients, PA occlusion occurred consistently in the early systolic phase regardless of the timing of balloon inflation. Mean Ppa, Ppc, and pulmonary arterial wedge pressure were 16.6, 11.8, and 7.6 torr. The ratio of venous to total resistance ranged from 0.37 to 0.54 (mean:0.46). We conclude that this technique is clinically feasible and valuable in precise definition of the instant of PA occlusion. By defining PA occlusion consistently, this technique can provide reliable Ppc estimation in the clinical settings. PMID- 8250303 TI - Airway management for patients with penetrating neck trauma: a retrospective study. AB - Airway management in patients with penetrating neck trauma is controversial. We reviewed the records of 107 patients with penetrating neck trauma from 1989 through 1991 for primary intubation technique, mechanism of injury, zone of injury, and structures injured. Six patients (6%) received a surgical airway as the primary choice, 89 (83%) had direct laryngoscopies after rapid sequence induction of anesthesia, eight (7%) had awake fiberoptic bronchoscopies, and four (4%) had awake blind nasotracheal intubation. The success rates for primary surgical were 100%, fiberoptic 100%, direct laryngoscopy 98%, and blind nasal 75%. Two (2%) of the patients in the direct laryngoscopy group required a secondary emergency surgical airway. One failed blind nasal attempt (25%) resulted in the patient's death due to loss of airway during a secondary attempt at an emergency surgical airway. A second patient died as a result of hemorrhage not related to airway management. Success rates were not statistically different with any of the four airway management techniques chosen primarily. However, the one death in the awake nasal intubation group and the technical and time constraints of fiberoptic intubation cause us to prefer rapid sequence induction of anesthesia with direct laryngoscopy or a primary surgical airway in patients with penetrating neck trauma who need an emergency airway. PMID- 8250304 TI - Pain on injection of propofol: modification by nitroglycerin. AB - The effect of applying nitroglycerin or placebo ointment to the back of the hand before venipuncture and injection of propofol was investigated in 60 ASA physical status I unpremedicated women. Eighteen patients (67%) pretreated with nitroglycerin experienced no pain compared with 10 (33%) in the placebo group. Eleven in the placebo group experienced moderate or severe pain during injection compared with only one in the active group. The time of onset of pain in more than half the subjects occurred 10 s or longer after commencement of injection, and, in more than half the patients, the site at which pain was felt was above the injection site (with three subjects experiencing it in the shoulder). No patient had a headache or experienced postural hypotension. We conclude that nitroglycerin ointment applied to the back of the hand before injection reduces the incidence of painful injection with propofol. PMID- 8250305 TI - Dose-response study of oral famotidine for reduction of gastric acidity and volume in outpatients and inpatients. AB - A relatively new H2-antagonist, famotidine, has become clinically available, but its effectiveness in decreasing gastric acidity and volume has not been compared in inpatients and outpatients. We reexamined the difference in gastric acidity and volume in inpatients and outpatients, and tested the effectiveness of different oral doses and dosage regimens of famotidine in reducing gastric acidity and volume in both groups of patients. Patients received either placebo or 20 mg or 40 mg of famotidine orally the night before surgery (HS) and on the morning of surgery (AM). One hundred forty-two inpatients and 180 outpatients were randomized to one of seven groups as follows: 1) placebo (HS)/placebo (AM); 2) 20 mg of famotidine (HS)/20 mg of famotidine (AM); 3) 20 mg of famotidine (HS)/placebo (AM); 4) placebo (HS)/20 mg of famotidine (AM); 5) 40 mg of famotidine (HS)/40 mg of famotidine (AM); 6) 40 mg of famotidine (HS)/placebo (AM); and 7) placebo (HS)/40 mg of famotidine (AM). We measured the gastric acidity and volume after induction of anesthesia and found no difference between the inpatients and outpatients, with or without famotidine. We found that famotidine given HS and AM or AM only was effective in reducing gastric acidity in both groups of patients, and that there was no difference between the 20-mg and 40-mg doses of famotidine. Gastric volume was not affected by any famotidine dose. PMID- 8250306 TI - Intrathecal sufentanil labor analgesia: the effects of adding morphine or epinephrine. AB - Intrathecal opioids can provide labor analgesia. We attempted to prolong the duration of intrathecal sufentanil analgesia by adding epinephrine or morphine. Forty-one healthy, term nulliparae with cervical dilation < 5 cm participated in this double-blind, randomized protocol. Using a combined spinal and epidural technique, we gave intrathecal injections of either sufentanil 10 micrograms, sufentanil 10 micrograms plus epinephrine 200 micrograms, or sufentanil 10 micrograms plus morphine 250 micrograms. At baseline and every 5 min for 30 min thereafter, we recorded arterial blood pressure and asked the patients to rate their pain, nausea, and pruritus on visual analog scales. The women continued to rate these variables every 30 min until they requested additional analgesia. They then received 10 mL of 0.25% bupivacaine via the epidural catheter, and rated the above variables every 30 min until they requested additional epidural drug injection. Both morphine and epinephrine prolonged the duration of sufentanil analgesia. Only morphine prolonged analgesia after the first dose of epidural bupivacaine. However, because women in the morphine group experienced significantly more side effects throughout the study period, we do not recommend intrathecal morphine for labor analgesia. PMID- 8250307 TI - Intrathecal sufentanil for labor analgesia--sensory changes, side effects, and fetal heart rate changes. AB - This study was designed to evaluate intrathecal (IT) sufentanil for labor analgesia with respect to sensory changes, side effects, and fetal heart rate (FHR) changes. In Phase I of the study, data regarding duration of analgesia and hemodynamic changes were obtained retrospectively from the labor and anesthetic records of 90 patients who had received IT sufentanil, 10 micrograms in 1 mL of saline, during active labor. In Phase II, an additional 18 parturients who received similar treatment were studied prospectively to document sensory, motor, and hemodynamic changes, as well as the incidence of side effects. In Phase I, analgesia occurred rapidly and lasted 124 +/- 68 min (SD); 19% of patients required no further analgesia before delivery. In Phase II, median time to onset of analgesia was 3 min (range 1-6 min) and mean duration of analgesia was 96 +/- 36 min. Decreased sensation to pinprick and cold occurred within 6 min extending from T4 to L4 (upper and lower median levels) in the majority of patients. All subjects requested additional analgesia within approximately 30 min of recession of sensory changes. Motor strength remained normal throughout. Hypotension (systolic blood pressure [BP] < or = 90 mm Hg or > 20% decrease in systolic BP) occurred in 14% and 11% of patients in Phase I and II, respectively. Perineal itching preceded analgesia in 95% of patients and all subjects experienced mild sedation. FHR changes occurred in 15% of cases but were not associated with adverse neonatal outcome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250308 TI - Postoperative pain: the effect of low-dose ketamine in addition to general anesthesia. AB - In a randomized, double-blind study, postoperative pain was assessed in 22 patients undergoing elective open cholecystectomy with two types of anesthesia: standardized general anesthesia (control group), and low-dose ketamine as an addition to the same method of general anesthesia, before surgical incision (ketamine group). After the operation we found that the time from the end of surgery to the first request for analgesic was longer in the ketamine group. Postoperatively, patients in both groups were treated with patient-controlled analgesia (PCA) in exactly the same way. The major difference in the study was the reduced dose requirement of morphine in the ketamine group compared with the control group after the operation. The mean dose of morphine given in patients of the control group during the first 24 h was 48.7 mg vs 29.5 mg in the ketamine group. Mean visual analog scale (VAS) and verbal rating scale (VRS) were higher in patients in the control group during the first 5 h after surgery (P < 0.02), but between 5 and 24 h after surgery VAS and VRS were not significantly different (P > 0.05). Our results indicate that postoperative pain can be decreased when ketamine in low doses is added to general anesthesia before surgical stimulation. PMID- 8250309 TI - Incremental dosing versus single-dose spinal anesthesia and hemodynamic stability. AB - We compared the hemodynamic stability after spinal anesthesia with two different dosing regimens in the elderly. Fifty patients, all older than 60 yr and scheduled for elective knee or hip surgery were assigned to two groups. After administration of 10 mL/kg of lactated Ringer's solution (RL) intravenously (i.v.) in the first group, we performed a continuous spinal anesthesia (CSA) by means of a 28-gauge catheter through which repetitive injections of 2.5-5 mg of plain bupivacaine 0.5% were given. In the other group, a single-dose spinal anesthesia (SS) with 20 mg of the same local anesthetic (LA) was carried out. Noninvasive mean arterial pressure (MAP), heart rate, and levels of analgesia were measured. To maintain MAP within 25% of initial value, the patients received additional i.v. fluids (RL) as first measure. When MAP could not be maintained despite hydration, incremental doses of ephedrine were given i.v. Six patients in the CSA group and 17 in the SS group developed a level of anesthesia higher than T6 (P < 0.01). In the SS group more fluid was needed (792 vs 388 ml) than in the CSA group (P < 0.01). Moreover, more patients of the SS group (11 vs 4) required ephedrine (P < 0.05). We conclude that CSA produces reliable and predictable analgesia for lower limb surgery with less need for correction of hemodynamic changes compared to SS. PMID- 8250310 TI - Regression of sensory anesthesia during continuous epidural infusions of bupivacaine and opioid for total knee replacement. AB - The epidural administration of morphine and fentanyl delay the regression of sensory anesthesia in postoperative patients receiving epidural bupivacaine. This study was performed to determine any differential effects of two lipid-soluble opioids upon regression of sensory anesthesia during coadministration with epidural bupivacaine. Forty-eight patients scheduled for total knee replacement underwent lumbar epidural catheterization and received 1.5% etidocaine with 1:200,000 epinephrine to establish sensory anesthesia to approximately T6 bilaterally. Patients were randomized by the investigational pharmacy to receive either: (a) bupivacaine without opioid (control) (n = 16), or (b) bupivacaine with 1 mg/mL of meperidine (n = 16), or (c) bupivacaine with 3 micrograms/mL of fentanyl (n = 16) in a double-blind fashion. Intraoperatively, 0.5% bupivacaine +/- opioid was administered by epidural infusion at a rate of 10 mL/h. Postoperatively, the bupivacaine concentration was decreased to 0.25% (+/- the same opioid), and the infusion rate was decreased to 4 mL/h. Pinprick sensory anesthesia and verbal numerical pain score were recorded each hour after surgery by a blinded investigator. For each patient, the study was considered terminated when the cephalad level of sensory anesthesia bilaterally decreased five dermatomal segments or the pain score reached "5" (moderate pain). Patients receiving epidural infusions of bupivacaine and meperidine had a significantly slower regression of sensory anesthesia and slower development of pain. There was no difference in the rate of regression of sensory anesthesia or the development of pain among patients receiving bupivacaine alone or bupivacaine with fentanyl. PMID- 8250311 TI - A dose-response study of orally administered clonidine as premedication in the elderly: evaluating hemodynamic safety. AB - Clonidine premedication in a dose of 5 micrograms/kg may be particularly well suited for elderly patients. To pursue this approach, sedation, intraocular pressure (IOP), and the hemodynamic profile of two doses of oral clonidine premedication were compared in 60 elderly patients, aged 65-82 yr, who underwent elective ophthalmic surgery under local anesthesia. Group 1 (n = 20) received placebo, Group 2 (n = 20) 150 micrograms of clonidine (2-2.5 micrograms/kg), and Group 3 (n = 20) 300 micrograms of clonidine (4-4.5 micrograms/kg) in a randomized, double-blind fashion. Decreases in mean arterial blood pressure were more pronounced and occurred earlier after 300 micrograms of clonidine (31.4 +/- 12.1%, P < 0.001) as compared to 150 micrograms of clonidine (18.1 +/- 10.9%, P < 0.001). Throughout the study, six patients (30%) in Group 3 (300 micrograms clonidine-treated group), but no patient in Groups 1 or 2, were treated at least once for hypotension (P < 0.05). Heart rate decreased significantly 18.5 +/- 8.1% (P < 0.001) only after 300 micrograms of clonidine. Clonidine 150 micrograms and 300 micrograms decreased IOP 32.1 +/- 14.3% (P < 0.001) and 47.8 +/- 17.2% (P < 0.001), respectively. After 150 micrograms of clonidine patients were significantly more sedated as compared to those given placebo (P < 0.01) but significantly less sedated than after 300 micrograms of clonidine (P < 0.01), where sedation persisted more than 6 h postoperatively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250312 TI - Pharmacokinetics and pharmacodynamics of rocuronium (Org 9426) in elderly surgical patients. AB - The effects of age on the pharmacokinetic and pharmacodynamic responses to rocuronium (Org 9426) were studied in 20 elderly (> 70 yr) and 20 younger control patients (< 60 yr) during N2O/O2, fentanyl anesthesia. The onset times were the same for both the elderly and younger control group, but the duration of action of rocuronium was significantly prolonged in the elderly patients. Elderly patients, when compared with the younger, also exhibited a significant decrease in plasma clearance (3.67 +/- 1.0 vs 5.03 +/- 1.5 mL.kg-1.min-1, mean +/- SD) and volume of distribution (399 +/- 122 vs 553 +/- 279 mL/kg, mean +/- SD). During the recovery phase of paralysis, no significant difference was seen in the log plasma concentration versus twitch tension response relationship between 20% and 80% paralysis in young and elderly patients receiving rocuronium. The differences in action of rocuronium between the elderly and younger groups can be fully explained by the observed differences in the distribution and elimination of rocuronium between the two groups. The decreased total body water and decreased liver mass which normally accompany aging are likely explanations for the pharmacokinetic changes found in the elderly in this study. We conclude that the action of rocuronium is prolonged in patients aged more than 70 yr because of decreased elimination of the drug. PMID- 8250313 TI - Pharmacokinetics and pharmacodynamics of pipecuronium in patients with cirrhosis. AB - To determine the effect of liver cirrhosis on the pharmacokinetics and pharmacodynamics of pipecuronium, the authors administered 100 micrograms/kg of pipecuronium intravenously to eight patients with liver cirrhosis and eight patients with normal liver and renal function undergoing elective abdominal surgery. All patients were anesthetized with thiopental (5-7 mg/kg), nitrous oxide (50-70% in oxygen), and fentanyl in repeated doses (2 micrograms/kg). Plasma concentrations of pipecuronium were determined by high-pressure liquid chromatography. A two-compartment open model was used for pharmacokinetic analysis. Thumb-elicited mechanical response to single-twitch ulnar nerve stimulation was also measured. Total plasma clearance did not differ between controls (2.96 +/- 1.05 mL.min-1.kg-1, mean +/- SD) and cirrhotics (2.61 +/- 1.16 mL.min-1.kg-1). Terminal elimination half-life was 111 +/- 46 min in controls and 143 +/- 25 min in cirrhotics. The total apparent volume of distribution at steady state did not differ between controls (350 +/- 81 mL/kg) and cirrhotics (452 +/- 222 mL/kg). The volume of the central compartment was not different between the two groups. The onset of neuromuscular blocking effect was longer in cirrhotics (233 +/- 112 s) (P < 0.05) than in controls (170 +/- 33 s). The clinical duration (injection until single twitch returned to 25%) was similar between the two groups: 167 +/- 41 min in controls and 165 +/- 48 min in cirrhotics. The authors conclude that hepatic insufficiency due to cirrhosis does not alter the pharmacokinetics and pharmacodynamics of pipecuronium (100 micrograms/kg). PMID- 8250314 TI - The effects of large-dose flumazenil on midazolam-induced ventilatory depression. AB - Flumazenil, a benzodiazepine antagonist, clearly reverses midazolam-induced sedation; reversal of ventilatory depression has not been as well demonstrated. Thirty-two subjects completed this randomized, double-blind, placebo-controlled study investigating the dose-response relationship and duration of flumazenil's effects on ventilatory depression and hypnosis induced by a continuous midazolam infusion. A computer-controlled infusion of midazolam was used to titrate the predicted midazolam plasma concentration to a level at which subjects were unresponsive to verbal commands and then to maintain that concentration. Measurements of ventilation and hypnosis were repeated at predetermined intervals: before midazolam administration, before test drug (flumazenil [1, 3, or 10 mg] or placebo), and 5, 30, 60, 120, and 180 min after test drug administration. Ventilation and tidal volume were measured during an isocapnic hyperoxia clamp at a PETCO2 of 46 mm Hg (VE46 and VT46, respectively). A pseudo rebreathing technique was used to measure the hypercapnic ventilatory response (HCVR) slope and ventilation intercept at a PETCO2 of 58 mm Hg (VE58). Midazolam reduced VE46, VT46, and VE58, as well as hypnosis scores, in all test drug groups. The reduction in HCVR slope, however, was significant only when all 32 subjects were considered in aggregate. All three doses of flumazenil reversed hypnosis and also reversed the reduction in VE46 and VT46 within 5 min. The reduction in VE58, however, was reversed less consistently. Flumazenil's effect on VE46 and VT46 lasted at least 30 min after 1 mg and at least 60 min after 3 mg, paralleling the effect of these doses on hypnosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250315 TI - Effects of halothane anesthesia on vasoconstrictor response to NG-nitro-L arginine methyl ester, an inhibitor of nitric oxide synthesis, in sheep. AB - This study tests the hypothesis that halothane-induced inhibition of the endothelium-derived relaxing factor/nitric oxide (EDRF/NO) pathway significantly contributes to cardiovascular performance and thus reduces the vasoconstrictor response to NO synthesis inhibitors in vivo. We determined the effects of the administration of the NO synthesis inhibitor NG-nitro-L-arginine methyl ester (L NAME) in chronically instrumented, halothane-anesthetized sheep and in awake control animals. Six sheep underwent halothane anesthesia (1.5 vol%) with mechanical ventilation. Five sheep were studied in the awake state with spontaneous breathing. Both groups received a bolus of L-NAME (25 mg/kg), followed 4 h later by L-arginine (300 mg/kg) to reverse the effects of L-NAME. L NAME administration caused a significant increase in pulmonary and systemic vascular resistance (P < 0.05) in both groups. However, L-NAME produced a sharp increase in mean arterial and pulmonary artery pressures only in the control group, whereas the pressor response in the halothane group was attenuated. Cardiac output, which was significantly lower after L-NAME administration in both groups, increased after L-arginine. The results suggest that halothane does not significantly alter the EDRF/NO-mediated effects on the vasculature but potentiates the cardiac depressant effect of L-NAME. PMID- 8250316 TI - The resting volume and compliance characteristics of the bronchial cuff of left polyvinyl chloride double-lumen endobronchial tubes. AB - The resting volume of a low-pressure, high-volume cuff is an important factor that determines the cuff compliance, because it is the maximum volume that maintains the low-pressure characteristics of the cuff. Modern polyvinyl chloride (PVC) double-lumen tubes (DLT) are designed with a low-pressure bronchial cuff to minimize the risk of bronchial damage. Maintenance of the low-pressure characteristics of this cuff, however, requires knowledge of how its resting volume and compliance vary between different DLT sizes and brands, and how the compliance changes when the cuff is inflated inside different-sized bronchi. We, therefore, measured the bronchial cuff pressure-volume relationship for each of the adult sizes, 35 Fr, 37 Fr, 39 Fr, and 41 Fr, of the Mallinckrodt, Sheridan, Rusch, and Portex left DLTs. The compliance of each cuff was characterized by: 1) the cuff resting volume, defined as the smallest cuff volume beyond which a 0.5 mL increase in volume resulted in more than 10 mm Hg increase in cuff pressure; and 2) the cuff pressure at 3-mL volume. Measurements were repeated for sizes 35 Fr and 41 Fr left Mallinckrodt DLTs with the bronchial cuff lying inside two canine left main-stem bronchi measuring 11 mm and 13 mm in diameter. The bronchial cuff resting volume ranged from 1.5 to 5.0 mL in different sizes and brands of DLTs, which could be explained by differences in cuff size between different manufacturers and between different DLT sizes of the same manufacturer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250317 TI - Use of the self-inflating bulb for detecting esophageal intubation after "esophageal ventilation". AB - This present investigation tests the efficacy of the self-inflating bulb for detecting esophageal intubation after intentional "esophageal ventilation" to mimic gastric insufflation after bag-and-mask ventilation. In 72 anesthetized patients, the trachea and esophagus were intubated with identical tubes. The efficacy of the bulb was tested by a second anesthesiologist before and after the delivery of three breaths at a tidal volume of 300-350 mL each. The pressures generated by the bulb connected to esophageally placed tubes were measured in 10 patients. In all patients, the second anesthesiologist reported no reinflation of the bulbs when connected to esophageally placed tubes and instantaneous reinflation when connected to tracheally placed tubes, thus correctly identifying the location of each tube. The mean negative pressure generated when compressed bulbs were connected to esophageally placed tubes was 55.4 +/- 1.2 mm Hg before esophageal ventilation and 59.0 +/- 0.68 mm Hg after esophageal ventilation. We conclude that insufflation of the stomach as a result of esophageal ventilation, to the extent demonstrated in this study, does not interfere with the effectiveness of the bulb in differentiating esophageal from tracheal intubation. PMID- 8250318 TI - Impairment of anesthesia task performance by laser protection goggles. AB - This study was designed to determine whether the wearing of colored goggles, commonly used during laser surgery, increased the time needed to perform simple tasks or the likelihood of medication errors. The Stromberg Dexterity Test and the Medication Sorting Task (MST), a test of our own design, were used to measure the speed and accuracy of identifying medications. Thirty anesthesia care providers performed the tests twice, once with colored goggles and once without. Ambient light during the test was controlled to approximate that in the operating room during laser surgery. Times required to complete the tests improved with repetition of the test. When colored goggles were worn, more time was needed to complete the tests. More errors, especially serious errors in medication sorting, occurred when goggles were worn than when they were not. The wearing of colored goggles in combination with low levels of illumination and stress due to time constraints, may increase the potential for medication errors. PMID- 8250319 TI - Hypokalemia and anesthetic implications. AB - Reductions in serum potassium influence myocardial cell excitability by increasing membrane potential, diastolic depolarization, duration of refractory period and action potential, and decreasing conduction velocity. Disturbances in electrolyte balance typically involve alterations in two or more cations whose effects can be additive or antagonist. Serum magnesium and calcium have been demonstrated to influence the electrophysiologic effects of potassium imbalances. The arrhythmogenic potential of hypokalemia is thought to result from electrical inhomogeneity, alterations in conduction, changes in automaticity and disturbances in sodium pump kinetics. Potassium balance is maintained by two separate, yet interrelated, systems: distribution and the balance between intake and excretion. Cell wall integrity, osmolality, hormones and acid-base balance influence the relative concentrations of potassium between the intracellular and extracellular compartments. Renal excretion is the major route of elimination and is affected by acid-base balance, potassium and sodium intake, urinary flow rates and mineralocorticoid states. Serum potassium is not an accurate reflection of total body potassium stores. Acute hypokalemia differs from chronic, for the former results in a change in only the serum potassium concentration, whereas the latter is accompanied by a reduction in both the total body stores and serum levels. The importance of intracellular and extracellular concentrations lies in their determination of the resting membrane potential and, therefore, membrane excitability. Although experimental studies have demonstrated an association between ventricular ectopy and hypokalemia, the clinical studies to date have reported conflicting results. The arrhythmogenic role of hypokalemia has been examined in ambulatory hypertensive patients with acute myocardial infarctions and those with magnesium deficiencies, the results of which have been difficult to interpret because of differences in study design, durations, and characteristics of study population. During general anesthesia, both experimental and clinical studies have failed to demonstrate an increase in the incidence of ventricular ectopy in hypokalemic patients. The common practice of acute repletion therapy or cancellation is not warranted based on the studies to date. Repletion therapy is neither inexpensive nor benign. In one study of 4921 hospitalized patients, the frequency of dangerous complications of oral and/or intravenous potassium therapy was approximately 1 in 175. Certain patients are susceptible to hyperkalemia, so commonly prescribed medication can promote elevations in serum potassium including indomethacin, amiloride, beta-adrenergic blocking agents, and angiotensin-converting enzyme inhibitors. Evaluation of hypokalemia should include identification of the etiology precipitating the electrolyte imbalance. Examinations should include the duration and severity of the depletion, history of past and present illness, current medications, and the presence of concurrent electrolyte disturbances.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8250320 TI - Perioperative management of children with mucopolysaccharidoses. PMID- 8250321 TI - Performance characteristics of Ultratherm fluid warmer. AB - Ultratherm Infusion Fluid Warmer Model 3703-1 (Ultratherm) is a disposable device approved by the Food and Drug Administration to warm fluids including blood and nutritional fluids. It has no temperature monitor or audible alarm and uses air activated chemical heating elements in direct contact with loops of intravenous (i.v.) tubing. We studied the warming of blood and saline by Ultratherm using intratubing thermocouple wire probes. Internal warmer chamber temperature reached 58 degrees C within 40 min of activation, and a plateau of approximately 65 degrees C reached in 1 h was sustained over the next 5 h. Free hemoglobin measurements on 47 degrees C and 53 degrees C blood samples obtained as blood exited the warmer at 30 mL/h demonstrated 0.6% and 7% hemolysis, respectively. Room temperature blood at 500 mL/h and refrigerated blood at 100 mL/h were warmed to approximately 36 degrees C. Refrigerated blood at 500 mL/h was not warmed above 19 degrees C. At flow rates of 500, 100, and 30 mL/h, room temperature saline reached 34 degrees C, 45 degrees C, and 52 degrees C, respectively. The unmonitored, high, internal warmer chamber temperatures preclude the use of Ultratherm with blood or any fluids potentially damaged by excessive heat, and its use should be limited strictly to clear fluids. The utility of the warmer, even with clear fluids, is further limited to room temperature fluids, to a time interval between 1 and 6 h following activation, and to flow rates between 100 and 500 mL/h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250322 TI - Penetrating cardiac trauma: a perioperative role for transesophageal echocardiography. PMID- 8250323 TI - Severe circulatory collapse immediately after pericardiocentesis in a patient with chronic cardiac tamponade. PMID- 8250324 TI - Tracheal stents: the Montgomery T-tube. PMID- 8250325 TI - Unanticipated difficult airway secondary to lingual tonsillar hyperplasia. PMID- 8250326 TI - Food anaphylaxis--an important cause of acute respiratory distress in infants. PMID- 8250327 TI - Neurotoxicity of lidocaine combined with mexiletine. PMID- 8250328 TI - An unusual case of back pain after epidural anesthesia: ureteral obstruction. PMID- 8250329 TI - Paranoid psychosis after intrathecal morphine. PMID- 8250330 TI - Potentiation of succinylcholine-induced phase II block by vancomycin. PMID- 8250331 TI - Relocation of a double-lumen tube during surgical dissection. PMID- 8250332 TI - Pharmacokinetic basis for local anesthetic infusions. PMID- 8250333 TI - Is uvular edema a complication of nasogastric tube placement? PMID- 8250334 TI - Transient neurologic toxicity after subarachnoid anesthesia with hyperbaric 5% lidocaine. PMID- 8250335 TI - Effect of "massage" on venous blood flow. PMID- 8250336 TI - Cardiac safety of ECT. PMID- 8250337 TI - Plasma drug efflux and pharmacokinetic models. PMID- 8250338 TI - The market has not failed! PMID- 8250339 TI - Vasoconstrictors in local anesthesia for dentistry. AB - Addition of a vasoconstrictor to a local anesthetic may have several beneficial effects: a decrease in the peak plasma concentration of the local anesthetic agent, increase in the duration and the quality of anesthesia, reduction of the minimum concentration of anesthetic needed for nerve block, and decrease of blood loss during surgical procedures. The addition of a vasoconstrictor to a local anesthetic may also have detrimental effects. A review of the literature indicates that vasoconstrictor concentrations in local anesthetics marketed for dental use in the United States are not always optimal to achieve the purposes for which they are added. In most cases, a reduced concentration of vasoconstrictor could achieve the same goal as the marketed higher concentration, with less side-effect liability. PMID- 8250340 TI - Limitations of pulse oximetry. AB - Pulse oximetry is a noninvasive, accurate, and safe method for the measurement of oxygen saturation during intravenous sedation or general anesthesia. Several factors should be considered with its use, since these variables will either alter the accuracy of the readings or may cause harm to the patient. These factors include changes in the strength of the arterial pulse, body movements, dyshemoglobinemias, plasma lipids and bilirubin, color interferences, venous pulsations, and several physical factors. Awareness of these variations will help the clinician become more knowledgeable in the use of the pulse oximeter. PMID- 8250341 TI - Anesthetic activity of the lipospheres bupivacaine delivery system in the rat. AB - The Lipospheres Bupivacaine Delivery System (bupivacaine-lipospheres) is a novel sustained-release local anesthetic preparation that has recently been made available for research purposes. This investigation compared the local anesthetic efficacy and safety of 2% bupivacaine-lipospheres, 0.5% bupivacaine plus 1:200,000 epinephrine, lipospheres plain, and physiologic saline following subcutaneous tail injection in the rat. A modified tail-flick paradigm was used to assess local anesthetic efficacy. Animals treated with 2% bupivacaine lipospheres or 0.5% bupivacaine with epinephrine displayed significant antinociception (P < 0.05) compared to saline or lipospheres plain with 5 min of injection. Bupivacaine with epinephrine had an anesthetic duration of 30 min, whereas 2% bupivacaine-lipospheres had a duration of 3 hr. The local anesthetic blockade produced by both active solutions was completely reversible. All animals gained weight normally during the 1-wk course of the study, and there were no signs of local tissue toxicity at the injection sites. We conclude that 2% bupivacaine-lipospheres is a safe and efficacious local anesthetic preparation in this particular animal model. It possesses an onset of action that is a rapid as 0.5% bupivacaine with 1:200,000 epinephrine, and a duration that is six times longer. PMID- 8250342 TI - Effect of head-down tilt on intracranial pressure and sagittal sinus pressure during general anesthesia in cats. AB - The effect of head-down tilt during general anesthesia on intracranial pressure (ICP) dynamics was examined in eight cats. Changes in lateral ventricular pressure (LVP), sagittal sinus pressure (SSP), and effective CSF pressure (ECSFP), which is the driving pressure of cerebrospinal fluid (CSF) absorption, were studied in association with a shift from the horizontal prone position to the 20 degrees head-down tilt position. Both LVP and SSP values were significantly (P < 0.01) increased at 10 min in the head-down tilt position as compared with the control position, remained elevated during the next 110 min, and returned to baseline when the horizontal position was restored. However, ECSFP (expressed by LVP - SSP) was not significantly different from the control value, because changes in LVP and SSP were similar. These results suggest that head-down tilt does not impair CSF absorption. PMID- 8250343 TI - Dental fear in Japan: Okayama Prefecture school study of adolescents and adults. AB - A total of 3,041 students and staff in middle school in Okayama Prefecture, Japan, were surveyed regarding dental fear. Over 88% reported fear, with 42.1% classified as having high fear. Almost 70% reported acquiring dental fear prior to junior high school. A majority reported being hurt at the last appointment. Delay of dental work was also reported for over 50% of the sample. Coping, pattern of physiological upset, nondental fears, and sex and age differences were also reported. Results suggest intervention is needed to address the major dental public health problems associated with dental fear. PMID- 8250344 TI - Experimental AIDS drug, d4T, now available to patients in critical need. PMID- 8250345 TI - Challenge procedures in detection of reactions to aspirin and nonsteroidal anti inflammatory drugs. PMID- 8250346 TI - Why adulterate theophylline? PMID- 8250347 TI - Tuberculosis and HIV infection: epidemiology, pathogenesis, and clinical aspects. AB - Human immunodeficiency virus infection has profoundly affected the epidemiology, natural history, and clinical presentation of TB. It has also contributed in part to the recent epidemic of MDR-TB in the US and significantly reduced the sensitivity (but not the specificity) of the tuberculin skin test. Further research is needed to determine the optimal strategy for chemoprophylaxis in HIV infected individuals, new methods of diagnosing TB infection and disease, and new drugs to control MDR-TB. Most importantly, however, further funding is necessary to improve compliance with therapy and chemoprophylaxis in order to regain control of this ancient scourge. PMID- 8250348 TI - Two siblings with recurrent infections. PMID- 8250349 TI - Minimizing acarians and house dust in the tropics. AB - Dust collection and mite isolation by Furumizo's method occurred in 33 homes on days 0 and 14. The first application of Paragerm AK (23 homes) or placebo aerosol (ten homes) was done after vacuuming on day 14. The second (final) spraying occurred on day 28, immediately after vacuuming. Vacuum collections without spraying were done on days 56 and 84 in this randomized double-blind trial. With Paragerm, dust volume fell to 80.3% of initial levels by day 28, remained low at 87.9% (P < .05) on day 56, and then rose to 115.9% by day 84. In control homes, comparative dust values were 89.0%, 103.7%, and 109.9%. With Paragerm, mite concentration decreased to 70.8% by day 28 (P < .01), to 65% at day 56 (P < .025), and continued to fall to 47.7% by day 84 (P < .05). With control aerosol, mite numbers rose compared with initial values: 119.1% at day 28, 227.2% at day 56, and 125.1% at day 84. No adverse effects were observed in Paragerm homes besides odor (70% complained). The two asthmatic occupants improved symptomatically and took less medication following the Paragerm. In conclusion, Paragerm, used over 40 years in Europe, is an effective, nontoxic acaricide in Hawaii, acting promptly and for at least 2 months. It was also associated with diminished dust volume, important in nondust allergy. PMID- 8250350 TI - Peak expiratory flow rates in healthy Saudi Arabian children living in Riyadh. AB - The peak expiratory flow rate (PEFR) was measured in 457 healthy Saudi schoolchildren (235 boys and 222 girls) aged 6 to 16 years, living in Riyadh, using two simple and commercially available airflow meters, Wright peak flow meter and the mini-Wright peak flow meter. All measurements were obtained in the standing position and the best of three trials was recorded. Standing height and age were used as independent variables. The regression equations for PEFR were determined for boys and girls separately. The boys had higher values than the girls at all heights. The difference was not significant. Both boys and girls had lower PEFRs as compared with European children and American children of different racial origin (P < .0001). When age was added to height as the second independent variable the Saudi boys and girls had significantly lower PEFRs than Swedish and British children (P < .0001). One explanation for these differences may be that the Saudi children on the average are shorter and have lower body weights than, eg, American children. The PEFR did not differ when using the Wright peak flow meter or the mini-Wright peak flow meter. These findings will serve as an important basis for preparing charts for normal PEFR values for Saudi children. PMID- 8250351 TI - Elevation of soluble IL-2 receptor and IL-4, and nonelevation of IFN-gamma in sera from patients with allergic asthma. AB - To determine whether the predominant in vivo activation of Th1 subset or Th2 subset in peripheral blood occurs in patients with allergic asthma, we measured serum levels of Th1 cell-derived interferon-gamma (IFN-gamma) and Th2 cell derived interleukin-4 (IL-4). In addition, serum levels of soluble interleukin-2 receptor (sIL-2R) released from activated T cells were measured to determine a potential source of these cytokines. The elevation of serum IL-4 levels and nonelevation of serum IFN-gamma levels were observed in patients with allergic asthma compared with normal control subjects. A significant correlation between serum IL-4 levels and serum sIL-2R levels was observed in these patients. These results suggest that there is allergen-induced in vivo activation of Th2-like cells in peripheral blood of patients with allergic asthma. PMID- 8250352 TI - Nonsteroidal antiinflammatory drug desensitization using flurbiprofen (Ansaid) AB - Nonsteroidal antiinflammatory drug (NSAID) and aspirin reactions remain commonplace. Specific recommendations and standardized approaches to the use of NSAIDs in patients with known hypersensitivity to these agents are evolving. Cross reactivity and sensitization are frequent within this group of agents. The advent of newer synthetic nonsteroidal antiinflammatory drugs may offer a means of lowering reaction rates. Flurbiprofen (Ansaid) is a new NSAID for the treatment of rheumatoid arthritis and osteoarthritis. We report successful flurbiprofen desensitization with incremental oral challenge in a patient using this agent. PMID- 8250353 TI - IgE-mediated allergy from vegetable allergens. AB - Allergy to fresh fruits and vegetables is IgE-mediated. Its main clinical features are local, such as oral pruritus and swelling of lips and tongue but systemic symptoms such as urticaria, asthma, or anaphylactic shock may occur. Clinical associations with allergic rhinitis due to cross-reactive antigens of pollens and foods are frequent. PMID- 8250354 TI - When a sigh is just a sigh . . . and not asthma. AB - Sighing is a normal physiologic response, expanding the lungs to vital capacity, usually followed by a prolonged expiratory phase. Sighing dyspnea is a condition that may be mistaken for asthma, and should be considered in the atypical cases. Recognizing sighing dyspnea at the onset may save patients from having to undergo extensive diagnostic evaluations and treatments. This condition, once identified, can often be easily treated by explaining the benign nature and giving reassurance to the patient. PMID- 8250355 TI - Passive transfer of IgE-mediated cutaneous reactivity in heterologous species. AB - The transfer of cutaneous hypersensitivity was performed in humans, monkeys, and dogs, using the Prausnitz-Kustner (PK) reaction. Quantitative endpoint titrations were used to study reactions resulting from sera from allergic humans, monkeys, and canines. IgE antibody from dogs sensitized human and rhesus monkey skin. IgE antibody from humans and rhesus monkeys did not sensitize canine skin. The Fc receptors on primate mast cells must have broader stoichiometric adaptation for IgE Fc receptors than the analogous Fc receptors on canine mast cells. PMID- 8250356 TI - Multi-Test skin testing in allergy: a review of published findings. PMID- 8250357 TI - Double-blind trial of pyridoxine (vitamin B6) in the treatment of steroid dependent asthma. PMID- 8250358 TI - [Axillary plexus block by simultaneous blockade of several nerves. I. Influence of the volume of the anesthetic solution]. AB - The influence of the volume of local anaesthetic solution on axillary blockade was investigated in a prospective randomized double-blind study including 120 patients presenting for upper limb surgery. A peripheral nerve stimulator was used to carry out the axillary block with a multiple injection technique. The musculocutaneous, radial, median and ulnar nerves were routinely stimulated. Four criteria were monitored: quality of sensory blockade, degree of motor blockade, time of onset and duration of anaesthesia. The patients were randomly allocated into two groups (n = 60 each). In the first, patients received 25 ml per m2 of body surface area of carbonated 1% lignocaine, 0.25% bupivacaine or a 1 to 1 mixture of both. Patients in the other group were given the same amount of local anaesthetic in half the volume, i.e. 12.5 ml.m-2 of carbonated 2% lignocaine, 0.5% bupivacaine or a 1 to 1 mixture of both these. Adrenaline (1 in 400,000) was added to half the anaesthetic solutions used in each group. The data demonstrated that, for a same amount of local anaesthetic, the larger volumes provided better quality sensory blockade than the smaller ones (p < 0.03). However, the volume of solution used affected neither the time of onset nor the duration of anaesthesia, nor the degree of motor blockade. It is concluded that, despite the use of a neurostimulator and simultaneous infiltration of several nerve trunks, the volume required to ensure a reliable degree of sensory block with the technique of axillary block is comprised between 40 and 50 ml (25 ml.m-2). PMID- 8250359 TI - [Axillary plexus block by simultaneous blockade of several nerves. II. Evaluation of lidocaine-bupivacaine combination]. AB - A mixture of carbonated lignocaine and bupivacaine for axillary blocks was assessed prospectively in a randomized double-blind study including 60 patients scheduled for upper limb surgery. A peripheral nerve stimulator was used to carry out the axillary block with a multiple injection technique. The musculocutaneous, radial, median and ulnar nerves were routinely stimulated. Four criteria were monitored: quality of sensory blockade, degree of motor blockade, time of onset and duration of anaesthesia. The patients were randomly assigned to three groups (n = 20 each). Patients received 25 ml per m2 of body surface area of either carbonated 1% lignocaine (group 1) or 0.25% bupivacaine (group 2) or a 1 to 1 mixture of both (group 3). Adrenaline (1 in 400,000) was added to half the anaesthetic solutions used in each group. The onset of anaesthesia was faster with the mixture (15 +/- 9 min) than will bupivacaine alone (23 +/- 14 min) (p < 0.02). It lasted longer in the mixture group (264 +/- 111 min) than in the lignocaine group (193 +/- 83 min). The longest duration was in the bupivacaine group (476 +/- 276 min) (p < 0.03). Adrenaline only affected significantly the duration of anaesthesia in the lignocaine group (252 min vs 135 min with and without adrenaline, respectively) (p < 0.03). In axillary blocks, the mixture of carbonated lignocaine and bupivacaine has a faster onset of action than bupivacaine alone, and a longer duration of action than lignocaine alone. Both agents provided a quality of sensory and motor blockade similar to that obtained with the mixture. PMID- 8250360 TI - [Local anesthesia of the finger through the flexor tendon sheath]. AB - A series of 350 cases of local anaesthesia of the finger using the flexor tendon sheath is reported. The patients were ranked ASA 1 or 2 and the youngest was 12 months old. This technique was used for day case surgery, such as nail trauma and tumours, wound exploration, tumour and cyst removal, tenolysis and neurolysis. The needle was inserted on the palmar aspect of the metacarpophalangeal joint, into the digital flexor tendon of the corresponding finger. Thereafter the needle was withdrawn very slightly in order to inject the local anaesthesia solution within the sheath, and not the tendon. The volumes were respectively 1 ml in children and 3 to 4 ml in adults of either 2% lignocaine or 0.5% bupivacaine without adrenaline. As palmar injections are painful a freezing spray solution was used prior to insertion of the needle. There was no anaesthesia of the dorsal digital nerves, as opposed to the findings of the author of the first series. This technique, which can be qualified as being easy and reliable, could be a valuable alternative for the ring or interdigital blocks, which carry a significant risk of vascular and neurological complications. PMID- 8250361 TI - [Preoperative normovolemic hemodilution using erythropheresis]. AB - Normovolaemic haemodilution was carried out by erythropheresis by the Blood Bank, 24 to 72 yours before surgery, using a plasmapheresis device. A special disposable haemodilution kit was used, with one plasma and three red cell collection bags and CPD as anticoagulant. Plasma was replaced by 4% albumin. One to 4 red cell packs with 75% haematocrit was obtained and stored for 10 days. This technique is indicated for surgery with an expected blood loss between 1,000 and 1,500 ml, or when the patient cannot be included in a preoperative blood donation programme. The contra-indications are the same as those of intentional normovolaemic haemodilution. This technique has been used in 101 patients due to undergo orthopaedic surgery. Compared with standard haemodilution bags, the advantages of this technique are the excellent asepsis of this product, its 10 days storage and high haematocrit, and the absence of anaesthetic drugs within the bags as well. This technique does not interfere with haemodynamic conditions during anaesthesia; it saves time for both anaesthetists and surgeons. In the authors' experience, this technique has in part replaced the more usual technique. PMID- 8250362 TI - [Comparative study of immunologic consequences of autotransfusion and homologous transfusion in lung cancer surgery]. AB - A study was carried out to determine the effects of transfusion with either autologous or homologous blood in 35 patients with lung cancer undergoing lung surgery. The patients were randomly allocated to two groups, "allo" receiving homologous blood (n = 10), and "auto" receiving autologous blood only (n = 13). A third control group included the patients (n = 12) who had not required any blood. In patients of "auto" group, two blood removals of 7 ml.kg-1 each were performed, provided their haemoglobin concentration was over 110 g.l-1 and their haematocrit over 34%. The following parameters were studied at t-1 (the day of inclusion), t0 (just before surgery and transfusion), t1 (one week after surgery), and t2 (one month later): haematocrit and haemoglobin concentration; IgG, IgA and IgM concentrations; polymorphonuclear and neutrophil leucocyte and lymphocyte numbers, lymphocyte subpopulations (CD4/CD8), activated natural killers (NK), polymorphonuclear leukocyte chemotaxis and chemoluminescence. Anaesthesia and surgery initiated initiated an early decrease in immunity in all three groups. The immune profile, and its time course in patients who received autologous blood only was similar to that of those who were not transfused at all. On the other hand, homologous blood transfusion seems to modify the recipient's non specific immune system. In group "allo", a decrease occurred in the number of auxiliary lymphocytes CD4, an increase in suppressor lymphocyte numbers CD8, with an increase in the CD4/CD8 ratio, and no increase in NK activity. These data suggest that autologous transfusion is well tolerated in cancer patients undergoing surgery. PMID- 8250363 TI - [Analgesia by sublingual buprenorphine in extracorporeal kidney lithotripsy]. AB - The efficacy of sublingual buprenorphine given for premedication was assessed in 287 consecutive patients scheduled for extracorporeal shock wave lithotripsy (ESWL). The patients were randomly assigned to three groups. They were given 45 min before starting ESWL 5 mg midazolam orally in group 1 (n = 97), and 5 mg midazolam orally and 0.2 mg sublingual buprenorphine in group 2 (n = 97), and 5 mg midazolam orally and 0.4 mg sublingual buprenorphine in group 3 (n = 93). Analgesia was assessed with a simple three step verbal scale: no pain, moderate pain treated with simple analgesics (Spasfon, Visceralgine), and exquisite pain requiring general anaesthesia. Sublingual buprenorphine provided good analgesia in 87% of group 2 patients, and 88% of group 3 patients (NS) whereas 70% of group 1 patients experienced no pain (p < 0.05). The incidence of side-effects during the 12 h following ESWL, such as nausea, vomiting and drowsiness, was higher in group 3 (31 patients) than in group 2 (13 patients) (p > 0.01). No clinical respiratory depression occurred, although 11 patients in group 3 complained of drowsiness. It is concluded that premedication with 0.2 mg of sublingual buprenorphine provides efficient analgesia for ESWL and few side-effects. PMID- 8250364 TI - [Analgesia after thoracotomy by extrapleural administration of continuous bupivacaine]. AB - This study was aimed to assess the efficiency and the side effects of a continuous administration of bupivacaine into the paravertebral space. Twenty patients, ranked ASA 2 or 3, with a mean age of 57.9 years, and having had a posterolateral thoracotomy for resection of lung tissue, were randomly assigned to one of two groups, B or C. At the end of the surgical procedure, a 22 gauge catheter was inserted into the paravertebral extrapleural space, at T4 levels As soon as pain occurred during recovery (T0), the patients were given two-hourly intravenous boluses of buprenorphine. The patients in group B were also given, through the paravertebral catheter, a 20 ml bolus of 0.25% bupivacaine, followed by a continuous steady rate infusion (10 ml.h-1). Group C patients were given normal saline in the same way. All patients could improve their analgesia with 0.05 ml boluses of buprenorphine given by an auto-analgesia pump (Pharmacia). The following parameters were assessed during the 72 h which followed the first injection: pain with a visual analogic scale, quality of sedation (5 grades), heart and breathing rate, systolic and diastolic blood pressure, arterial blood gases. In group B, plasma bupivacaine concentrations were measured throughout the infusion, and for an 8-hour period after its end. The statistical analysis included 15 patients only, as the catheter had moved into the chest cavity in the other 5. Analgesia was qualified to be adequate by all patients, but there was no statistically significant difference in the amounts of self-administered buprenorphine between groups B and C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250365 TI - [Postoperative respiratory function and cholecystectomy by laparoscopic approach]. AB - Open cholecystectomy is associated with characteristic changes in pulmonary function showing a restrictive pattern. Laparoscopic cholecystectomy without opening of the peritoneal cavity could be an alternative in reducing postoperative respiratory dysfunction. Having given their informed consent, 13 healthy ASA1 patients (age: 41 +/- 18 yrs) undergoing laparoscopic cholecystectomy were enrolled in this study, in order to assess their postoperative pulmonary function tests (forced vital capacity [FRC], forced expiratory volume [FEV1], functional residual capacity [FRC]) before operation (T0) and 4 h (T4), 24 h (T24), 48 h (T48) after surgery. Anaesthesia technique was the same associating propofol-atracurium-fentanyl, 50% N2O/O2. Ventilation was adapted to maintain end-tidal carbon dioxide pressure up to 30-35 mmHg. Postoperative analgesic regimen consisted of paracetamol-ketoprofen. Mean length of surgery was 84 +/- 15 min; mean duration of anaesthesia was 110 +/- 24 min. An immediate and harmonious restrictive breathing pattern developed postoperatively. Postoperative FVC measured 65% (T4), 63% (T24), 72% (T48) of preoperative function (p < 0.025); postoperative FEV1 measured respectively 60, 66 and 75% of preoperative function (p > 0.001), without change in FEV1/CV and FRC; a significant hypoxia occurred (T0: 86 mmHg, T4: 80 mmHg, T24: 75 mmHg, T48: 81 mmHg [p < 0.05]). Laparoscopic cholecystectomy resulted in less postoperative respiratory dysfunction than conventional cholecystectomy, as previously reported; this restrictive pattern observed without changes in FRC was similar to that following lower abdominal surgery. PMID- 8250366 TI - [Use of fiberoptic bronchoscope for difficult intubation in maxillofacial surgery]. AB - This retrospective study analyzed the cases of difficult intubations carried out with a fiberoptic bronchoscope between March 1984 and May 1989. During this period, 222 such procedures were attempted in 131 male and 68 female patients. All cases were diagnosed at the preoperative visit. A fiberoptic bronchoscope was used as a guide under topical nasal and laryngeal anaesthesia, together with appropriate benzodiazanalgesia, in order to maintain spontaneous breathing. The nasotracheal route was used in 218 cases. Successful intubation was achieved in 219 cases (98.6%), 209 of them (95.4%) within fifteen minutes. Of the three failures, only one was unpredictable (inability to withdraw the fiberoptic bronchoscope from the endotracheal tube). In the two others, the technique was subsequently found to have been wrongly indicated: major nasopharyngeal secretions occluded the fiberoptic device in the first one, and an anatomically compromised airway led to apnoea under sedation in the other. A narrow nasotracheal passage, spreading oedema, bloody secretions or coughing gave rise to technical difficulties in 39 cases (17.5%). There was one case of regurgitation without any drawback. Benzodiazanalgesia was responsible for two cases of respiratory depression. This technique was otherwise very satisfactory, patients being cooperative and frequently having amnesia of intubation. The indications for the use of a fiberoptic bronchoscope were: insufficient oral opening, orofacial obstacles to laryngoscopy, and cases where laryngoscopy had to be avoided. The main drawbacks of this technique were the cost and fragility of fiberoptic device as well as operator efficiency. The anaesthesiologists involved in the present series performed ten easy intubations each with the fiberoptic bronchoscope, and routinely used the teaching eye-piece.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250367 TI - [Effects of isoflurane and halothane on motor evoked potentials in the rabbit]. AB - The effects of isoflurane or halothane on motor evoked potentials (MEPs) were assessed and compared in the rabbit, in order to contribute to devise a rigorous human anaesthetic protocol to be used for monitoring of MEPs in corrective spinal surgery. Ten black adult New Zealand rabbits were anaesthetised twice at a month interval, once with isoflurane and once with halothane. Once a control cortical stimulation had been carried out after the animal had breathed pure oxygen for three minutes, the following concentrations of anaesthetic agent were given for 3 min each: respectively 0.3 vol %, 0.5 vol % and 1 vol % of isoflurane, and 0.5 vol % and 1 vol % of halothane. Cortical stimulation was carried out every minute. The signs of anaesthesia (diameter decrease of the pupil, eye covered by the nictating membrane), muscle relaxation (ears drop) and breathing rate were recorded. MEPs were recorded 1, 2, 3 and 5 minutes after the end of anaesthesia. Isoflurane had a stronger effect on MEPs than halothane. The effect was more pronounced on amplitude than on latency. MEPs remained present whatever the concentration of halothane. In 70% of cases, MEPs, discontinued with isoflurane, more rapidly, more deeply, and for a longer time. Mean latency was more constantly increased in the isoflurane than in the halothane group. The effect of volatile halogenated anaesthetics on mean latency of MEPs seemed to be more delayed than that on amplitude. One should also take into account an individual sensitivity, it is concluded that the interpretation of MEPs during anaesthesia with volatile halogenated agents should be carried out with caution. PMID- 8250368 TI - [Prophylactic antibiotherapy using cefapirin in the surgery of duodenal ulcer: a randomized clinical trial]. AB - A randomized clinical trial of antibiotic prophylaxis in patients undergoing gastroduodenal surgery for stenosing or bleeding duodenal ulcer was carried out from February 1990 to February 1991. Seventy-two patients were randomly assigned to either Group I (n = 33) and given 1 g of cefapirine intravenously every eight hours, four times, starting at induction of anaesthesia; or to Group II (n = 39), and not given any antibiotic. The surgical dressing was changed on the third postoperative day, and thereafter every two days in patients with fever. The efficiency of prophylaxis was assessed on the incidence of superficial or deep primary infection, and the length of hospital stay. Primary wound sepsis occurred in 7 Group II patients, and in none in Group I (n = 0.01). The results for those patients with stenosing ulcer (n = 53) were also significant: there was no primary wound infection in Group I (n = 25), whereas five occurred in Group II (n = 28) (p = 0.03). Hospital stay was shorter in Group I than in Group II (7 vs 9.5 days). Antibiotic prophylaxis also seemed to be efficient in patients with bleeding duodenal ulcers (n = 19), but their group was too small to reach levels of statistical significance. Again hospital stay was shorter in Group I than in Group II (8.5 vs 12.2 days). The efficiency of antibiotic prophylaxis was therefore confirmed for patients undergoing gastroduodenal surgery for stenosing duodenal ulcer. PMID- 8250369 TI - [A rare cause of obstruction of the intubation tube]. AB - A case is reported of endotracheal tube obstruction due to impaction of a turbinate. This complication occurred during a first attempt of nasotracheal intubation in an ASA 1 18-year-old patient. Once the tube had been inserted into the trachea, manual ventilation was impossible. The diagnosis was made on removal of the completely obstructed tube. Differential diagnosis was sudden severe bronchospasm or a herniated cuff. Anaesthesiologists should be aware of this rare complication when carrying out nasotracheal intubation. Means of prevention are proposed. PMID- 8250370 TI - [Temporary relief from pain in the phantom limb after spinal anesthesia using a combination of bupivacaine and clonidine]. AB - Two cases are reported of patients with phantom limb pain after lower limb amputation and requiring surgery of their stump. The stumps were revised and the remaining femoral shaft shortened. Both had spinal anaesthesia for this procedure, with an association of bupivacaine and 150 micrograms of clonidine. In one patient, the phantom pain did not resume for one month, and in the other for three days. The return of pain was preceded by the sensation of a phantom limb. These cases suggest that alpha 2 adrenergic agonists could play a major part in the treatment of phantom limb pain. PMID- 8250371 TI - [Rhabdomyolysis in severe acute asthma]. AB - A case is reported of a 74-year-old man who developed rhabdomyolysis during treatment for status asthmaticus. This patient had a history of asthma. He was admitted after a cardiac arrest during a severe asthma attack. Continuous muscle relaxation (4 mg.h-1 of pancuronium for 10 days; total dose 960 mg) was required to carry out mechanical ventilation. Bronchodilators and high doses of steroids (300 mg.day-1 of methylprednisolone for 10 days) were also given. After the administration of muscle relaxant had been discontinued, the patient displayed a severe, predominantly proximal, quadriplegia as well as a raised blood creatinine kinase concentration. The urine was brown coloured over a 24 h period. Rhabdomyolysis was confirmed by muscle biopsy. The patient recovered over a period of one month. This case is discussed in the light of some other similar reports in the literature. It seems that the combination of muscle relaxant with high doses of steroids is to be incriminated. PMID- 8250373 TI - [A new laryngoscope with a multiple-use blade]. PMID- 8250372 TI - [Rhabdomyolysis after ORL surgery in prolonged lateral position]. AB - Two cases are reported of upper limb rhabdomyolysis occurring after prolonged ENT cancer surgery, the patient being part of the time in the right lateral position, with the left forearm outstretched. Surgery consisted of a vertical hemilaryngectomy with immediate parascapular free graft reconstruction in a 48 year-old man (10 h of surgery, with 6 h in a lateral position) and the surgical removal of a neoplasm involving the mouth floor and larynx in a 62-year-old man including parascapsular free graft reconstruction (10 h of surgery, with 7 h in a lateral position). In the early postoperative period, the patients complained of severe pain in the left forearm. There was a tense painful swelling of the forearm, combined with an increase in creatinine kinase plasma concentration, and myoglobinaemia. The mechanism involved was most likely a compression of the forearm muscles, together with prolonged surgery and the patient position. The part played by the state of the patients is discussed. Diagnosis must be made as soon as possible, as the only efficient treatment consists of an early fasciotomy together with the administration of alkali to avoid renal failure. PMID- 8250374 TI - [Myocardial postpartum hypokinesia: role of salbutamol]. PMID- 8250375 TI - [Peritoneal mucormycosis and fluconazole]. PMID- 8250376 TI - [Consensus conference. Antibiotic prophylaxis in surgery in adults. French Society of Anesthesia and Intensive care (11-12 December 1992)]. PMID- 8250377 TI - Biomedical Engineering Society, 25th Anniversary, 1993 annual fall meeting. Memphis, Tennessee, October 21-24, 1993. Abstracts. PMID- 8250378 TI - Evaluation of limb symmetry indices, using ground reaction forces in healthy dogs. AB - Limb symmetry was evaluated by measuring ground reaction forces in 2 groups of normal-gaited dogs at a trot. Data were collected from 2 groups of 21 dogs trotted at dog/handler velocities of 1.25 to 1.55 m/s and 1.85 to 2.05 m/s, respectively. Of these dogs, 9 participated in both groups to allow comparison of data at both velocities. Additionally, 16 of the dogs in group 1 were measured in 2 directions of movement to determine whether directional dependence was present. Collected data were then applied to 3 described symmetry indices. Each index was easy to calculate, but all had limitations. A major limitation was variation in magnitude of ground reaction forces measured between the different axes and the effect of this variation on precision of the derived indices. Vertical ground forces provided the most consistent symmetry indices, in part because of their large magnitude. The indices indicated that no dog had perfect right-to-left symmetry during a trotting gait. Statistical differences were not found in any of the measurements of directional dependence. Likewise, comparing symmetry data in dogs trotted at both velocities indicated no significant differences in any axis. However, further analysis of the data revealed the actual amount that a variance attributable to right-left limb variation was negligible. Most of the variance was attributable to trial variation. Thus, the aforementioned indices, which use nonconsecutive footfall methods to evaluate limb symmetry, actually measure principally trial variation and not limb-to-limb variation. PMID- 8250379 TI - Endogenous creatinine clearance measurement of glomerular filtration rate in dogs. AB - Renal clearance procedures were performed on adult mixed-breed dogs with a wide range of renal function. Endogenous creatinine clearance was computed after analyzing plasma and urine for creatinine by use of 2 methods, PAP and kinetic Jaffe. For 20-minute clearance procedures, [14C]inulin clearance was measured simultaneously with endogenous creatinine clearance. For 111 twenty-minute clearance procedures performed on 24 dogs, [14C]inulin clearance was highly correlated with creatinine clearance for both methods of creatinine analysis (R2 = 0.979 for [14C]inulin-PAP; R2 = 0.943 for [14C]inulin-Jaffe). The absolute values for PAP and [14C]inulin clearance were nearly the same (PAP-to-[14C]inulin clearance ratio = 1.03 +/- 0.08), but those for Jaffe clearance were substantially less than those for [14C]inulin clearance (Jaffe-to-[14C]inulin clearance ratio = 0.88 +/- 0.10). The Jaffe-to-[14C]inulin clearance ratio was inversely correlated with degree of renal function (R2 = 0.464), whereas the PAP to-[14C]inulin clearance ratio was not correlated with degree of renal function (R2 = 0.060). Thus, Jaffe-determined creatinine clearance varied, in relation to [14C]inulin clearance, depending on degree of renal function. In 4 clinically normal dogs, 20-minute and 24-hour sample collections analyzed by use of the PAP method gave clearance values significantly greater, for both periods, than did Jaffe analyses. The PAP-determined creatinine clearance values were less than, but not significantly different from 20-minute exogenous creatinine clearance values determined 10 days after 24-hour collections.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250380 TI - Evaluation of the indirect fluorescent antibody test for diagnosis of Babesia gibsoni infections in dogs. AB - We determined the extent of the serologic cross-reactivity in the indirect fluorescent antibody (IFA) test for Babesia gibsoni, and the optimal cut-off titer for seropositivity in the test. The lowest titer to B gibsoni detected in a dog with naturally acquired clinical babesiosis was 1,280, and 7 of 12 dogs had titer between 10,240 and 20,480. Two experimentally infected normosplenic dogs developed high titer (40,960 to 81,920) to B gibsoni, and the same sera reacted in IFA tests for B canis (titer < or = 640), Toxoplasma gondii (titer < or = 2,560), and Neospora caninum (titer < or = 10,240). Dogs that were experimentally infected with B canis and T gondii had titer < or = 160 to B gibsoni. Dogs that were experimentally infected with N caninum had titer (80 to 10,240) to N caninum, but failed to have serologic reactivity to B gibsoni. Serologic titer of healthy dogs from Australia, a country where B gibsoni is not known to exist, was < or = 160 to B gibsoni. On the basis of these findings, a cut-off titer of 320 was considered to be appropriate for serodiagnosis of B gibsoni in dogs with clinical signs of babesiosis. A more conservative titer of 1,280 was established as the cut-off titer for seroepidemiologic studies based on relative costs and benefits of false-positive results and failure to isolate B gibsoni parasites after splenectomy and immunosuppression from a clinically normal dog with B gibsoni titer of 640.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250381 TI - Use of a DNA probe to detect the intracellular organism of proliferative enteritis in swine feces. AB - A method of extracting bacterial DNA from swine feces was developed and used in a molecular assay for the presence of ileal symbiont (IS) intracellularis, formerly known as the Campylobacter-like organism associated with swine with proliferative enteritis. Hybridization with a digoxigenin-labeled, IS intracellularis-specific probe detected the presence of IS intracellularis at a concentration of 10(7) organisms/g of feces. This method was sufficient to detect IS intracellularis in the feces of swine with experimentally induced and naturally acquired infection. Results of the hybridization were in agreement with those from histologic postmortem examination. PMID- 8250382 TI - Immune responses and protection against infection and abortion in cattle experimentally vaccinated with mutant strains of Brucella abortus. AB - Twenty-four 10-month-old Polled Hereford heifers were inoculated SC with live cells of one of the following strains of Brucella abortus: S19 delta 31K (n = 4), S19 delta SOD (n = 4), RB51 (n = 4), and strain 19 (n = 6); controls (n = 6) were given saline solution. Heifers given the deletion mutants S19 delta 31K and S19 delta SOD, and those given strain 19 developed antibody responses to B abortus and cutaneous reactions to brucellin. Heifers given strain RB51 did not develop antibodies that reacted in the standard tube agglutination test, but sera reacted in tests, using an antibody dot-blot assay containing RB51 antigen. The S19 delta 31K and S19 delta SOD strains of B abortus isolated from lymph node tissue after vaccination did not differ genetically from the master stock strain. All heifers were bred naturally at 16 to 17 months of age, and were challenge-exposed intraconjunctivally with virulent B abortus strain 2308 during the fifth month of pregnancy. All vaccinated heifers were protected (ie, none aborted and none had B abortus isolated from their tissues after parturition). Calves born from vaccinated dams were free of B abortus. Antibody responses in heifers after challenge exposure were an indicator of immunity. All 5 control heifers (nonvaccinated) developed serum antibodies after challenge exposure; 3 aborted, and 1 delivered a small, weak calf at 8.5 months of gestation. Thus live mutant strains of B abortus can induce protective immunity when given at 10 months of age, and strain RB51 is a strong candidate for further testing. PMID- 8250383 TI - Leukocyte mobilization to skin lesions in dogs. AB - A suction blister technique was used in 10 healthy dogs to remove the epidermis from the dermis in a standardized way. Collection chambers were attached to these skin windows and filled with autologous serum to attract exudative neutrophils. The chambers were emptied by fine-needle aspiration at 4-hour intervals and were refilled with serum for 24 hours after the last aspiration. The collected cells were counted, differentiated, and stained, using the trypan blue dye-exclusion method to determine cell viability. Multiple skin biopsy specimens obtained during the procedure were examined histologically. The chamber fluid collected after 24 hours was cultured for bacteria. Increasing numbers of viable neutrophils were collected during the 24-hour period from the induced skin windows. In all but 1 dog, sufficient viable neutrophils could be collected to perform further functional tests in vitro. Our conclusion is that this technique might be useful to study chemotaxis in vivo and to perform functional tests on exudative neutrophils. PMID- 8250384 TI - Clinical and endoscopic study to estimate the incidence of distal respiratory tract infection in thoroughbred foals on Ontario breeding farms. AB - Respiratory tract infections are prevalent in foals, yet the frequency with which the distal airways are affected in clinical episodes of respiratory tract disease has not been evaluated to our knowledge. The objective of the study was to determine the incidence of distal respiratory tract infection (DRTI) in foals on a sample of Thoroughbred breeding farms (n = 10) in Ontario. In a pilot study, clinical criteria commonly used to select foals for antimicrobial treatment (detection of abnormal lung sounds, plus nasal discharge, cough, fever, tachypnea, and/or lethargy) were found to segregate foals with and without endoscopically confirmed DRTI. Mucopurulent exudate and bronchial erythema were observed more frequently (P < 0.005), bronchial lavage total cell count and neutrophil concentration were significantly (P < 0.005) higher, and intracellular cocci were recovered significantly (P < 0.01) more often from bronchial lavage samples of affected foals (n = 8) than of controls (n = 8). These clinical criteria were used to identify cases in a cohort of Thoroughbred foals (n = 219) from May 1 to October 30, 1991. Case morbidity adjusted for clustering was 82 +/- 5% (95% confidence limits, 72 to 92%). Most (74%) episodes of clinical DRTI were detected in July and August, and equal numbers were detected before (53%) and after (47%) weaning of foals. Of 178 cases, 66 (48%) were selected at random for endoscopy and bronchial lavage. Grade-II pharyngeal lymphoid hyperplasia was observed commonly (60% of foals); auditory tube diverticulum (guttural pouch) discharge was observed in 18 of 86 (21%) foals, and guttural pouch infection was confirmed in 6 of 7 foals examined endoscopically.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250385 TI - Microbiologic changes during antimicrobial treatment and rate of relapse of distal respiratory tract infections in foals. AB - Despite the high incidence of distal respiratory tract infection of undetermined cause on farms, to our knowledge, the microbiologic effects of conventional antimicrobial treatment for this condition have not been studied. We evaluated the possible pathogenic role of bacterial isolates from the distal airways of foals with clinical respiratory tract disease, by correlating changes in their numbers (increase or decrease) with clinical, endoscopic, and pulmonary cytologic signs of disease resolution during treatment with antimicrobial drugs. We also determined qualitative changes in in vitro antimicrobial susceptibility of bacterial isolates after 7 days of treatment and relapse rate of foals. Significant (P < 0.05) decrease in the numbers of an isolate in the airways was considered strong evidence of a pathogenic role in this disease syndrome. Foals with endoscopically confirmed distal respiratory tract infection (DRTI; n = 65) were selected at random for treatment (n = 56) or nontreatment (n = 9), and bronchial lavage specimens were cultured and evaluated cytologically before and after 7 days of treatment with trimethoprim-sulfamethoxazole (TMS) and a beta lactam drug (penicillin, ampicillin, or sulbactam-ampicillin), the standard treatment in all foals. The effect of treatment was to abruptly reduce the clinical (nasal discharge, cough, adventitious lung sounds) and cytologic signs of airway infection. Severity of disease in nontreated foals, however, did not change or did worsen over time. Reduction in the frequency and numbers of Streptococcus zooepidemicus isolated during treatment supported a causal role for this organism in the clinical syndrome observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250386 TI - Association of microbiologic flora with clinical, endoscopic, and pulmonary cytologic findings in foals with distal respiratory tract infection. AB - Undifferentiated distal respiratory tract disease (nasal discharge, cough, pneumonia) in foals (1 to 8 months old) is a burdensome economic problem on breeding farms; yet, the infective agents associated with these episodes have not been well described. Possible causes of these episodes of illness were investigated by culturing specimens of proximal and distal airways of clinically diseased foals (n = 101), prior to any treatment, for aerobic and anaerobic bacteria and viruses (rhinoviruses, equine arteritis virus, equine herpesvirus subtype 1 [EHV-1], influenza virus, and adenovirus). Pairs of sera (n = 47) were examined for antibodies to influenza A virus, equine subtypes 1 and 2, EHV-1, and adenovirus antigens, and sera obtained from foals during acute infection were examined for antibodies (by agar gel immunodiffusion [AGID]) to equi factor antigens of Rhodococcus equi. Viruses were not isolated from the proximal (swab) or distal (bronchial lavage) airway specimens in foals, and only 2 of 47 randomly selected foals seroconverted to EHV-1. Serotiters to the other viruses were low and frequently decreasing between samples, which was compatible with maternally derived antibody. Streptococcus zooepidemicus was the predominant isolate from bronchial lavage specimens (88/101 cases), accompanied by alpha-hemolytic streptococci (8 cases), Bordetella bronchiseptica (13 cases), Staphylococcus epidermidis (9 cases), and other organisms in lesser frequency. Only Str zooepidemicus was recovered significantly (P < 0.05) more often in cases than in controls. The AGID test was found useful to detect foals with presumed exposure to R equi, but positive tests results did not correspond well with bacterial culture results; positive AGID results were recorded in 34% of culture-negative foals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250387 TI - Production and characterization of VP4/VP7 reassortant swine rotaviruses derived from Gottfried and OSU parental strains. AB - The ability of viral glycoproteins (VP) VP4/VP7 reassortant swine rotaviruses (RV) to induce cross-neutralizing antibody against parental serotypes was investigated in guinea pigs. Using selective culture conditions, we produced 10 reassortant viruses that contained gene segment 4 of the OSU RV strain and gene segment 9 of the Gottfried RV strain. These reassortant RV grew to high titer in cell culture and were neutralized by monospecific antisera against both parental RV strains. The reassortant RV were chemically inactivated with binary ethylenimine, adjuvanted with aluminum hydroxide, and used to produce antisera in guinea pigs. The hyperimmune antisera had high neutralization titer against both parent RV strains. These results indicate that several of the reassortant RV may be capable of inducing neutralizing antibodies to VP4 and VP7 and may have future use as bivalent vaccine strains. PMID- 8250388 TI - Antigenic and genetic analysis of a recently isolated H1N1 swine influenza virus. AB - Hemagglutinins (HA) of H1N1 swine influenza viruses isolated in the United States have remained antigenically and genetically conserved for many years. In contrast to such conservation, the HA of A/Swine/Nebraska/1/92 (Sw/Neb) could readily be distinguished from those of contemporary porcine viruses. Twenty-eight amino acid mutations differentiated the HA of Sw/Neb and A/Swine/Indiana/1726/88, the most recent H1N1 swine influenza virus for which HA sequence data were available. Among these differences were mutations at potential asparagine-linked glycosylation sites and charge changes at many residues. The Sw/Neb virus also could be differentiated from other swine influenza viruses in hemagglutination inhibition assays with monoclonal antibodies to recent H1 swine HA. Nonetheless, overall sequence analysis of the HA and the nucleoprotein genes of Sw/Neb indicated that this virus was more closely related genetically to classic H1N1 swine influenza viruses than to H1N1 avian or human viruses. Infection of swine with Sw/Neb under experimental conditions induced clinical signs and lesions typical of swine influenza. However, affected swine in the field had high, persistent fevers, but relatively mild signs of respiratory tract disease. This study indicated that an antigenically and genetically novel variant of swine influenza virus was detected in the United States. PMID- 8250389 TI - Efficacy of Pasteurella haemolytica subunit antigens in a goat model of pasteurellosis. AB - The effectiveness of Pasteurella haemolytica biovar A, serovar 1 (Ph1) subunit vaccines was tested in goats, using challenge exposure by transthoracic injection. Twenty-two weanling male Spanish goats were randomly allotted to 4 groups. Six goats were given 2 transthoracic injections into the lung 18 days apart with live Ph1 impregnated in agar beads (positive controls). Six goats were not given injections (negative controls). Five goats were given 2 transthoracic injections into the lung 18 days apart with 4.6 mg of cytotoxin in agar beads. The remaining 5 goats were given 2 IM injections, 18 days apart, into the thigh with 4.6 mg of cytotoxin emulsified in incomplete Freund's adjuvant. Twenty-four days after the second injection, all goats were challenge-exposed to live Ph1 by transthoracic injection into the lung, and 4 days later, all goats were euthanatized and necropsied. Serum neutralizing anticytotoxin titer was measured throughout the experiment. Mean volume of consolidated lung tissue was 0.38 cm3 for the positive control group, 32 cm3 for the negative control group; 19 cm3 for the cytotoxin-lung group; and 88 cm3 for the cytotoxin-adjuvant-IM group. Only the positive control group was protected from Ph1 challenge exposure. The Ph1 cytotoxin subunit vaccine alone appeared to be ineffective, and the anticytotoxin titer was not correlated with protection. In a separate trial, 32 weanling male Spanish goats were randomly allotted to 5 groups. Each was given 2 transthoracic injections into the lung 22 days apart. Six goats were given Ph1 cytotoxin impregnated into agar beads; 6 were given Ph1 lipopolysaccharide impregnated in agar beads; 6 were given Ph1 capsule impregnated in agar beads. Six goats were given agar beads only (negative controls), and 6 were given live Ph1 impregnated into agar beads (positive controls). Twenty days after the second injection, all goats were challenge-exposed to live Ph1 by transthoracic injection into the lung, and 4 days later, all goats were euthanatized and necropsied. Mean volume of consolidated lung tissue was 0.14 cm3 for the positive control group, 7.59 cm3 for the negative control group, 11.21 cm3 for the cytotoxin group, 10.19 cm3 for the lipopolysaccharide group, and 1.6 cm3 for the capsule group. Again, only injection of live Ph1 (positive controls) induced solid protection; however, the capsule subunit vaccine induced partial protection against challenge exposure in this trial. Lipopolysaccharide and cytotoxin subunit vaccines were ineffective in protecting goats against challenge exposure with live Ph1. PMID- 8250390 TI - Effects of dietary protein and calorie restriction in clinically normal cats and in cats with surgically induced chronic renal failure. AB - To determine the effects of long-term dietary protein restriction in cats with chronic renal failure (CRF), 4 healthy adult cats and 7 cats with surgically induced CRF were fed a high-protein (HP, 51.7% protein) diet and 4 healthy adult cats and 7 cats with surgically induced CRF were fed a low-protein (LP, 27.6% protein) diet for 1 year. Cats with induced CRF that were fed the LP diet had reduced serum urea nitrogen concentrations, despite lower glomerular filtration rates, compared with cats with CRF fed the HP diet. Despite five-sixths reduction in renal mass, reduced glomerular filtration rate, and azotemia, 13 of the 14 cats with induced CRF retained the ability to concentrate urine and produced urine with a specific gravity > 1.035. Cats fed the HP diet consumed significantly more calories than did cats fed the LP diet, presumably because the HP diet was more palatable. As a result of the lower caloric intake in cats fed the LP diet, these cats were protein and calorie restricted, compared with cats fed the HP diet. Cats fed the HP diet weighed significantly more than did cats fed the LP diet. Mean hematocrit and mean serum albumin concentration were significantly lower in control cats and in cats with CRF fed the LP diet, compared with control cats and cats with CRF fed the HP diet. Hypokalemia developed in 4 of 7 cats with CRF fed the HP diet (containing 0.3% potassium); hypokalemia did not develop in control cats fed the same diet or in cats with CRF fed the LP diet containing 0.4% potassium. Excessive kaliuresis, hypomagnesemia, and metabolic acidosis did not appear to contribute to the hypokalemia. Subsequent supplementation of the HP diet with potassium gluconate prevented hypokalemia in cats with CRF. PMID- 8250391 TI - Association of host cell mitochondria with developing Toxoplasma gondii tissue cysts. AB - Ultrastructure of the interactions of host cell mitochondria with developing Toxoplasma gondii tissue cysts was examined in cultured cells, using transmission electron microscopy of infected cells and rhodamine 123 (a mitochondria-specific vital fluorescent dye) staining of isolated tissue cysts. Structurally mature T gondii tissue cysts were observed as early as 2 days after inoculation of cultured cells. During development of T gondii, host cell mitochondria were observed surrounding the parasitophorous vacuole membrane. Mitochondria became flat and elongated in the vicinity of the parasitophorous vacuole membrane. These mitochondria were also closely associated with T gondii tissue cysts. Incubation of tissue cysts from cultured cells and tissue cysts from mouse brains with rhodamine 123 revealed fluorescence of the tissue cyst wall in living specimens. Incubation of tissue cysts with 10 microM rotenone caused diminished fluorescence of the tissue cyst walls, and 100 microM rotenone caused complete inhibition. Mouse RBC, and tissue cysts fixed in 100% methanol did not fluoresce after exposure to rhodamine. Tissue cysts in 9 isolates of T gondii from mouse brains were examined, using rhodamine 123, and the tissue cysts walls of all isolates fluoresced, indicating no isolate effects. Our results indicate that host cell mitochondria may be closely associated with the tissue cysts of T gondii in cell cultures and in mice. PMID- 8250392 TI - Serologic and parasitologic responses of domestic chickens after oral inoculation with Toxoplasma gondii oocysts. AB - Four-week-old chickens were inoculated orally with 1,000 or 100,000 oocysts of the ME-49 or GT-1 strain of Toxoplasma gondii, and their antibody responses were measured, using the direct modified agglutination test, latex agglutination test, indirect hemagglutination test, ELISA, and the Sabin-Feldman dye test. Antibodies against T gondii were detected by use of the modified agglutination test and ELISA within 2 weeks of oocyst inoculation, and antibodies persisted until termination of the study by postinoculation day 68. The latex agglutination test was insensitive in detecting T gondii antibodies, and antibodies were not detected by use of the dye and indirect hemagglutination tests. Of tissues bioassayed in mice for tissue cysts by pepsin digestion of individual organs of chickens on postinoculation day 68, tissue cysts were found in the brain of all 5, heart of 3, and leg muscles of 2, but not in the liver and breast muscles. None of the birds developed clinical toxoplasmosis. PMID- 8250393 TI - Evaluation of the microcirculation of the equine small intestine after intraluminal distention and subsequent decompression. AB - Effects of intraluminal distention (25 cm of H2O, 120 minutes) and subsequent decompression (60 minutes) on intramural vascular patterns of the small intestine was evaluated in 7 anesthetized horses. Intraluminal distention (25 cm of H2O, 120 minutes) was created in 2 jejunal segments in each horse. Experimental and control segments were removed either immediately after the experimental period or after 60 minutes of decompression. The vascular system of experimental and control jejunal segments was lavaged with NaCl, then was injected with a blue colored radiopaque medium for microangiography or with a diluted methyl methacrylate for scanning electron microscopy of microcorrosion vascular casts. After angiographic evaluation, tissue sections were prepared for light microscopic evaluation to assess vascular filling and tissue morphology. The distended segments had short villi, which were separated by expanded crypts, and had mesothelial cell loss, neutrophil infiltration, and edema in the seromuscular layer. The number of perfused vessels was significantly (P < 0.05) decreased in the seromuscular layer and, to a lesser extent, in the mucosal layer of the distended segments, compared with controls. After decompression, the morphologic lesions progressed in mucosal and serosal layers and the number of observed vessels increased in all intramural layers; however, vascular density did not return to the predistention state. These results identify altered intramural vascular patterns in the equine jejunum during luminal distention and subsequent decompression. PMID- 8250394 TI - Evaluation of the microcirculation of the equine jejunum and ascending colon after ischemia and reperfusion. AB - Intramural vascular patterns of the jejunum and colon were evaluated during ischemic strangulation obstruction (ISO, 70 minutes) and subsequent reperfusion (60 minutes) in 7 adult anesthetized horses. Microvasculature of experimental and control segments was described by comparison of results from microangiography, light microscopy, and scanning electron microscopy of vascular replicas. Experimental and control segments with isolated vascular arcades were removed either immediately after the experimental period or after 60 minutes of reperfusion. Blood was flushed from the vascular system by use of isotonic NaCl, and the segments were divided. Half of each segment was perfused with a modified radiopaque medium for microangiographic evaluation, and half was perfused with dilute methyl-methacrylate to create a vascular replica to be studied by scanning electron microscopy. Microangiographic section also were evaluated for histologic changes. Microvasculature of jejunal control segments and all colon segments was similar to described normal microvasculature of the equine jejunum and ascending colon. In jejunal ISO segments, intramural perfusion was redistributed away from the mucosa. In the villi, the central arteriole was short and convoluted and the subepithelial capillaries were not filled. The submucosal vessels and crypt capillaries were congested, compared with those of controls, and the serosal vessels were not filled in the ischemic segments. Histologic grade II-III mucosal lesion was seen in jejunal ISO segments. Reperfused jejunal segments had a transmural hyperemic response, and previously unfilled capillaries were observed in all intestinal layers. After reperfusion, the mucosal lesion progressed to grade III-IV and a cellular infiltrate and edema formation were observed in the serosa. The intramural vasculature of the ischemic and reperfused colon remain unchanged. Minimal histologic damage was observed in the colon after 70 minutes of ISO or after 60 minutes of reperfusion. PMID- 8250395 TI - Recovery of horses from inhalation anesthesia. AB - To study behavioral and cardiopulmonary characteristics of horses recovering from inhalation anesthesia, 6 nonmedicated horses were anesthetized under laboratory conditions on 3 different days, with either halothane or isoflurane in O2. Anesthesia was maintained at constant dose (1.5 times the minimum alveolar concentration [MAC]) of halothane in O2 for 1 hour (H1), halothane in O2 for 3 hours (H3), or isoflurane in O2 for 3 hours (I3). The order of exposure was set up as a pair of Latin squares to account for horse and trial effects. Circulatory (arterial blood pressure and heart rate) and respiratory (frequency, PaCO2, PaO2, pHa) variables were monitored during anesthesia and for as long as possible during the recovery period. End-tidal percentage of the inhaled agent was measured every 15 seconds by automated mass spectrometry, then by hand-sampling after horses started moving. Times of recovery events, including movement of the eyelids, ears, head, and limbs, head lift, chewing, swallowing, first sternal posture and stand attempts, and the number of sternal posture and stand attempts, were recorded. The washout curve or the ET ratio (end-tidal percentage of the inhaled agent at time t to end-tidal percentage of the inhaled agent at the time the anesthesia circuit was disconnected from the tracheal tube) plotted against time was similar for H1 and H3. The slower, then faster (compared with halothane groups) washout curve of isoflurane was explainable by changes in respiratory frequency as horses awakened and by lower blood/gas solubility of isoflurane. The respiratory depressant effects of isoflurane were marked and were more progressive than those for halothane at the same 1.5 MAC dose. During the first 15 minutes of recovery, respiratory frequency for group-I3 horses increased significantly (P < 0.05), compared with that for the halothane groups. For all groups, arterial blood pressure increased throughout the early recovery period and heart rate remained constant. Preanesthesia temperament of horses and the inhalation agent used did not influence the time of the early recovery events (movement of eyelids, ears, head, and limbs), except for head lift. For events that occurred at anesthetic end-tidal percentage < 0.20, or when horses were awake, temperament was the only factor that significantly influenced the nature of the recovery (chewing P = 0.04, extubation P = 0.001, first stand attempt P = 0.008, and standing P = 0.005).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8250396 TI - Modulation of bronchial responsiveness in horses by phenylbutazone and furosemide. AB - Effects of phenylbutazone (PBZ) and furosemide (FUR) on the respiratory tract of horses were evaluated, focusing on bronchial responsiveness. Four healthy Thoroughbreds were used and data were analyzed by use of a Latin square design. Histamine provocation tests (0.5, 1, 2, and 4 micrograms/min, i.v.) were done: (1) without prior treatment with PBZ or FUR, (2) 30 minutes after administration of PBZ (8 mg/kg, i.v.), (3) 1 hour after administration of FUR (1 mg/kg, i.v.), and (4) after administration of PBZ plus FUR. Pulmonary function tests (dynamic compliance, resistance, respiratory frequency, and tidal volume) and heart rate were monitored throughout the experiments. Phenylbutazone did not influence basal pulmonary function test results, whereas FUR caused a significant (P < 0.05) increase in dynamic compliance and decrease in resistance. Histamine infusion resulted in a dose-dependent decrease in dynamic compliance and a dose-dependent increase in resistance, respiratory frequency, and heart rate. Phenylbutazone administration significantly (P < 0.05) attenuated most of the changes induced by histamine, whereas FUR had less protective action. Administration of PBZ plus FUR before administration of histamine was less effective than administration of PBZ alone. PMID- 8250397 TI - Cardiorespiratory effects of intravenous bolus administration and infusion of ketamine-midazolam in dogs. AB - Twelve healthy dogs were used to determine the cardiorespiratory effects of i.v. administered ketamine (10 mg/kg of body weight) and midazolam (0.5 mg/kg). Half the dogs received a ketamine-midazolam combination (K-M) as a bolus over 30 seconds and the other half received the K-M as an infusion over 15 minutes. Induction of anesthesia by use of K-M was good in all dogs. Ketamine-midazolam combination as a bolus or infusion induced minimal cardiorespiratory effects, except for significant (P < 0.05) increases in mean heart rate and rate-pressure product. The increase in heart rate was greater in dogs of the infusion group. Mild and transient respiratory depression was observed in dogs of both groups immediately after administration of K-M, but was greater in dogs of the bolus group than in dogs of the infusion group. Duration of action of K-M for chemical restraint was short. Salivation and defecation were observed in a few dogs. Extreme muscular tone developed in 1 dog after K-M bolus administration. PMID- 8250398 TI - Cardiovascular effects of intravenous bolus administration and infusion of ketamine-midazolam in isoflurane-anesthetized dogs. AB - Cardiovascular effects of IV administered ketamine (10 mg/kg) and midazolam (0.5 mg/kg) were determined in 12 healthy isoflurane-anesthetized (1.7% end-tidal concentration) dogs. Six dogs received a ketamine-midazolam combination (K-M) as a bolus over 30 seconds and 6 dogs received K-M as an infusion over 15 minutes. Ketamine-midazolam combination as a bolus and an infusion caused early significant (P < 0.05) reductions in mean systemic blood pressure, cardiac index, and stroke index, which returned to baseline values near the end of the study. Heart rate decreased significantly (P < 0.05) in dogs of the infusion group and returned to the baseline value near the end of the study. One dog died after K-M bolus administration. Mean maximal decreases from baseline for systemic blood pressure, cardiac index, and stroke index were significantly (P < 0.05) greater in dogs of the bolus group than in dogs of the infusion group; therefore, cardiovascular effects of K-M after infusion were less severe than those after bolus. Base excess and pHa decreased significantly (P < 0.05) in the infusion group, although similar changes occurred in both groups. Four dogs were maintained with 1.7% end-tidal isoflurane to determine temporal effects of isoflurane; these dogs did not receive K-M. Increases in heart rate, cardiac index, stroke index, and left and right ventricular stroke work indexes were significant (P < 0.05) at various sample collection intervals, particularly during the later stages of the study. Isoflurane anesthesia effectively blocked the cardiostimulatory properties of K-M.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250399 TI - Influence of anesthetic regimens on the perioperative catecholamine response associated with onychectomy in cats. AB - Plasma catecholamine concentrations in response to onychectomy were examined in 27 cats receiving different anesthetic regimens. Each cat was anesthetized with a dissociative-tranquilizer combination, and onychectomy was performed on 1 forefoot. One week later, each cat was anesthetized with the same dissociative tranquilizer combination plus either butorphanol or oxymorphone, and onychectomy was performed on the other forefoot. Four treatment groups were studied: tiletamine-zolazepam and tiletamine-zolazepam-butorphanol combinations were administered to group-1 cats, ketamine-acepromazine and ketamine-acepromazine butorphanol combinations were administered to group-2 cats, tiletamine-zolazepam and tiletamine-zolazepam-oxymorphone combinations were administered to group-3 cats, and ketamine-acepromazine and ketamine-acepromazine-oxymorphone combinations were administered to group-4 cats. All drug combinations were administered IM. Central venous blood samples were drawn for catecholamine analysis after injection of drug(s), after onychectomy, and 1, 2, and 4 hours after injection. Tiletamine-zolazepam alone or tiletamine-zolazepam-butorphanol prevented epinephrine release for 2 hours after injection of drug(s). Norepinephrine concentration increased significantly (P < 0.05) from baseline after onychectomy for tiletamine-zolazepam-butorphanol and at 4 hours for tiletamine-zolazepam and tiletamine-zolazepam-butorphanol. After onychectomy, there was no difference in epinephrine values between tiletamine-zolazepam and tiletamine-zolazepam-oxymorphone. Ketamine-acepromazine prevented increases in norepinephrine and epinephrine concentrations for up to 2 hours after surgery. Addition of butorphanol to ketamine-acepromazine decreased norepinephrine values immediately after onychectomy. Addition of oxymorphone to ketamine-acepromazine resulted in lower epinephrine values 4 hours after surgery. PMID- 8250400 TI - Technique for prolonged, minimally invasive monitoring of intragastric pH in ponies. AB - Intragastric pH monitoring was investigated in ponies. In cadaver stomachs, close contact with the mucosa led to high pH readings if nonweighted electrodes were used. However, pH recorded by weighted electrodes was markedly less affected by mucosal contact (P < 0.001). The latter were used for subsequent trials. In vivo, high correlations were found between pH recorded by weighted electrodes with or without a wire guard to prevent mucosal contact (correlation, r = 0.866; P < 0.001). Readings from each correlated well with those from simultaneous gastric aspirates (r = 0.774 and r = 0.807, respectively; P < 0.001 for both correlations). Plain electrodes recorded more highly variable (temporally heterogeneous) pH than did guarded electrodes. In vitro, trials using equine gastric fluid indicated that this resulted from greater responsiveness of the plain electrode. In vivo, episodes of nearly neutral pH were a common feature, and high pH correlated with intensely yellow-green, neutral fluid in the stomach (rank correlation, rho = 0.626; P < 0.01). Concentration of bile acids did not correlate with pH or color score (rho = -0.158 and rho = 0.076, respectively). Causes of the episodes could include salivary influx, duodenogastric reflux, and variable gastric acid secretion. Pentagastrin infusion (0.6 microgram/kg of body weight/h) reduced intragastric pH (P = 0.018), but episodes of neutrality still occurred. Experiments in fed ponies indicated possible existence of a stable pH gradient, from neutral dorsally to heterogeneous and more acidic ventrally. Care was required in the rational choice of summary variables for expression of monitored pH data. Of the frequency distributions of 3 summary variables assessed in this study (mean, median, and percentage of data > pH 4), only that of the mean approached normality. Thus, use of the mean may allow analysis by parametric statistical methods. Intragastric pH monitoring was found to be a useful technique. Episodes of increased pH were often identified. These may represent episodic duodenogastric reflux. PMID- 8250401 TI - Effect of constant administration of a gonadotropin-releasing hormone agonist on reproductive activity in mares: induction of ovulation during seasonal anestrus. AB - The potential of a gonadotropin-releasing hormone (GnRH) agonist (goserelin acetate), delivered constantly for 28 days via a subcutaneous depot, to induce ovulation in seasonally anestrous mares, was investigated. Two experiments were conducted, in which a range of doses (30 to 240 micrograms/mare/d) was examined. Mares were selected on the basis of lack of substantial follicular development (follicle diameter < 20 mm determined ultrasonically) and low serum concentrations of luteinizing hormone (LH) and progesterone. Constant administration of the GnRH agonist-induced ovulation in anestrous mares, but a dose-response relation was not observed. Furthermore, with identical doses tested in consecutive or alternate years, considerable variation was observed in the ovulatory response. In general, ovulation in all treated mares was accompanied by increased circulating concentrations of LH and a decrease in follicle-stimulating hormone values. Ovulation was preceded by an increase in estradiol and LH concentrations. In mares in which ovulation did not occur, concentration of LH increased during agonist treatment, whereas that of follicle-stimulating hormone either increased or did not change. It was concluded that constant administration of GnRH agonists may induce ovulation in mares during seasonal anestrus; however, percentage of mares ovulating and the lack of reproducibility of effect indicate that this approach is inappropriate for use as a reliable method to manipulate breeding activity in commercial broodmares. PMID- 8250402 TI - Effect of constant administration of a gonadotropin-releasing hormone agonist on reproductive activity in mares: preliminary evidence on suppression of ovulation during the breeding season. AB - During the breeding season, the effect of constant administration of an agonist analog of gonadotropin-releasing hormone (GnRH; goserelin acetate) on reproductive activity of mares was determined. Twenty-four mares undergoing estrous cycles were allocated at random to 6 groups (n = 4/group) and, on May 29 (day 0), received no treatment (group 1, controls), 120 micrograms (group 2), 360 micrograms (group 3), 600 micrograms (group 4), or 1,200 micrograms (group 5) of GnRH agonist/d for 28 days via a depot implanted subcutaneously. The final group of mares (group 6) was treated with 120 micrograms of GnRH agonist/d for 84 days (3 occasions at 28-day intervals). During a pretreatment period (April 19 to May 29) and for 90 days after initiation of GnRH agonist treatment, follicular development and ovulation were monitored by transrectal ultrasonography of the reproductive tract at 2- to 3-day intervals. On each occasion a blood sample was collected for determination of luteinizing hormone (LH) and progesterone. Estrous behavior was monitored by teasing of mares with a stallion. Initiation of agonist treatment was random, relative to the stage of the estrous cycle, and all mares ovulated within 11 days before or after implantation. In 3 of 4 nontreated control mares, estrous cycles were observed throughout the study, with interovulatory intervals ranging from 18 to 26 days. In the remaining mare, concentration of progesterone was high after asynchronous double ovulation during the pretreatment period, suggestive of persistent corpus luteum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250403 TI - Measurement of pulmonary diffusing capacity for carbon monoxide and functional residual capacity during rebreathing in conscious thoroughbreds. AB - A rebreathing method for measurement of pulmonary diffusing capacity for carbon monoxide (DLCO) and functional residual capacity (FRC) was evaluated in conscious horses. Horses were manually ventilated through an endotracheal tube, using a custom-made syringe filled with a gas mixture containing 18-carbon monoxide (18CO) and helium (He). The 18CO and He concentrations were continuously monitored by use of a mass spectrometer connected to the rebreathing circuit. Values for DLCO and FRC were calculated from changes in the concentration of these 2 gases. In 11 Thoroughbreds, mean (+/- SD) DLCO was 330.3 +/- 56.9 ml.min 1 x mm of Hg-1, and FRC was 20.21 +/- 3.35 L. Body weight normalization yielded mean (+/- SD) values of 0.652 +/- 0.114 ml.min-1 x mm of Hg-1 x kg-1 for DLCO, and 39.9 +/- 6.4 ml.kg-1 for FRC. PMID- 8250404 TI - Influence of hydration state on renal functions of dogs. AB - Clinically normal dogs were evaluated in states of dehydration, euhydration, and after fluid administration to determine effects of hydration state on renal clearance values. Endogenous creatinine, exogenous creatinine, and [14C]inulin clearances, were determined to measure glomerular filtration rate (GFR); in some experiments p-aminohippurate clearance was determined to measure renal plasma flow. Dehydration caused significant (P < 0.05) decrease in clearance values, compared with euhydration, and clearance values during euhydration were significantly (P < 0.05) less than values obtained after a single gavage with water (30 ml/kg of body weight). Sustained administration of 3 fluid regimens was evaluated for effects on clearance values (treatment A = 30 ml of lactated Ringer's solution/kg/h; treatment B = 30 ml of water/kg by gavage hourly; treatment C = 10 ml of glucose:lactated Ringer's solution/kg/h). All regimens of fluid therapy caused significant P < 0.05), progressive increases in GFR, but treatment C resulted in the most stable GFR values. Increases in clearance values were associated with positive fluid balance; the rate of fluid administration was greater than the rate of urine formation. Data from 285 GFR determinations on 85 dogs were evaluated retrospectively. For each determination, three 20-minute urine collections were made beginning 40 minutes after 30 ml of water/kg was given by gavage. Values between collections were significantly (P < 0.05) different, but varied by < 3%. Comparison of methods for measurement of GFR indicated that endogenous creatinine clearance and [14C]inulin clearance were highly correlated (R2 = 0.82), but mean clearance values were markedly different (mean +/- SEM, 28.70 +/- 0.01 and 37.07 +/- 1.29 ml/min, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250405 TI - Circulating concentration of dexamethasone in healthy dogs, dogs with hyperadrenocorticism, and dogs with nonadrenal illness during dexamethasone suppression testing. AB - Concentration of dexamethasone was determined in plasma or serum samples from dogs after i.v. administration of a low dose (0.01 mg/kg of body weight) or high dose (0.1 mg/kg) of dexamethasone. On the basis of history, clinical signs of disease, and degree of cortisol suppression in response to dexamethasone, dogs were assigned to these groups: healthy dogs, dogs with nonadrenal illness, and dogs with hyperadrenocorticism. Four hours after administration of the low dose of dexamethasone, concentration of the steroid was reduced (P < 0.05) in dogs with hyperadrenocorticism, compared with healthy dogs, but not compared with values from dogs with nonadrenal illness. By 8 hours after dexamethasone administration, values were similar across groups. Dexamethasone concentration 4 and 8 hours after high-dose administration was similar between healthy dogs and dogs with hyperadrenocorticism. Concentration of dexamethasone 4 and 8 hours after its administration overlapped after the 2 doses. For example, in 11 of 66 dogs from all groups, concentration measured 4 hours after the low dose was greater than the minimal concentration determined in the 18 dogs given the high dose. These data indicate that dexamethasone metabolism may be altered in dogs with hyperadrenocorticism, and that individuals may have appreciable variability in dexamethasone clearance. Such variability provides a possible explanation for false-positive and false-negative results associated with dexamethasone suppression testing in dogs. PMID- 8250406 TI - Biomechanical comparison of 1-screw and 2-divergent pin internal fixations for treatment of slipped capital femoral epiphysis, using specimens obtained from immature dogs. AB - Biomechanical strength and stiffness of 2 fixation treatments used to repair acute slipped capital femoral epiphysis were evaluated in bone specimens form immature dogs. A servohydraulic testing machine was used to create slipped capital femoral epiphysis in 7 pairs of femurs by shearing the capital femoral epiphysis along the physis in a craniocaudal direction. The slip was reduced and repaired with one 3.5-mm-diameter screw placed in lag fashion or 2 double pointed, 1.6-mm (0.062 inch)-diameter smooth pins and retested. Strength and stiffness of each intact femur (which served as the control) and repaired femur were compared. Results of the study indicated that the failure strength of 2-pin fixation was significantly (P < 0.05) weaker than its control and the 1-screw fixation. There was no significant difference between failure strength of the 1 screw fixation and its control. The stiffness of 1-screw and 2-pin fixations was not significantly different, compared with each other, but was significantly (P < 0.05) less, compared with their respective controls. PMID- 8250407 TI - Effects of vena caval banding in experimentally induced multiple portosystemic shunts in dogs. AB - Effects of vena caval banding on portal venous and vena caval hemodynamics were examined in 6 control dogs and in 10 dogs that had undergone attenuation (banding) of the abdominal part of the caudal vena cava and had dimethylnitrosamine-induced multiple portosystemic shunts (PSS). Additionally, indocyanine green (ICG) extraction and clearance after infusion to steady state were used to calculate hepatic plasma flow in these dogs. Sixteen dogs were randomly assigned to 2 groups: control (n = 6) or diseased (n = 10). Diseased dogs were administered dimethylnitrosamine (2 mg/kg, PO, twice weekly) until multiple PSS developed, as assessed by results of clinical laboratory tests, ultrasonography, and hepatic scintigraphy. Shunts were confirmed visually at celiotomy and by contrast portography. Venous pressures (caudal vena caval, portal, and hepatic) were recorded before and after vena caval banding for up to 7 days in dogs from both groups. Peritoneal cavity pressures were recorded in all dogs after closure of the body wall. To determine ICG extraction and clearance, a bolus injection of ICG (0.5 mg/kg, i.v.) was administered, followed by steady state infusion of 0.097 mg/min. Extractions and clearances of ICG were measured, and from these, hepatic plasma flow rates were determined immediately before and after banding and at 6 hours, 48 hours, and 7 days after banding. The gradient (caudal vena caval pressure within 1 to 2 mm of Hg of portal pressure) between caudal vena cava and portal venous pressures established at banding was maintained after the first hour in both groups. Caudal vena cava pressures established at banding were maintained throughout the study, with the exception of the first hour in diseased dogs. Extraction ratios were higher in control dogs at all times, except at 48 hours. Clearance was higher in control dogs at all times. Hepatic plasma flow did not differ between groups, except immediately after banding, when flow was greater in diseased dogs, and differences were not found over time in either group. This study indicated that vena caval banding in this model of experimentally induced multiple PSS increases and maintains caudal vena cava pressure, relative to portal venous pressure (after the first hour) for 7 days, and that calculated hepatic plasma flow is not persistently improved by vena caval banding. PMID- 8250408 TI - Effects of intra-articular chlorhexidine diacetate lavage on the stifle in healthy dogs. AB - Eight dogs were determined to be orthopedically normal on the basis of prelavage physical examination, stifle radiography, synovial fluid analysis, and force plate analysis (peak vertical force normalized for body weight, and time on the force plate). Each dog had 1 stifle randomly assigned to be lavaged with 100 ml of a commercially available 0.05% (w/v) chlorhexidine diacetate solution, and the contralateral stifle was lavaged with lactated Ringer's solution. Difference was not detected between the chlorhexidine diacetate and lactated Ringer's solution treated joints, with regard to results of synovial fluid analysis and clinical lameness evaluations on days 4 and 8 after lavage. Chlorhexidine diacetate caused a more intense synovitis than did lactated Ringer's solution, as determined by histologic evaluation of synovial membrane specimens after necropsy on day 8; however, a difference in the intensity of toluidine blue staining of articular cartilage was not found between treatments. Chlorhexidine diacetate, as a 0.05% (w/v) solution, cannot be recommended as a joint lavage fluid until the duration of inflammatory changes in the synovial membrane are determined or until the chemical constituents of chlorhexidine diacetate causing the synovitis can be identified and removed. PMID- 8250409 TI - Nurses win greater role under Clinton health plan. PMID- 8250410 TI - RN career security is ANA goal for reform. PMID- 8250411 TI - Consumers willing to see a nurse for routine 'doctoring'. PMID- 8250412 TI - Kristine Gebbie urges nurses to be active in war against AIDS. Interview by Kathryn Scott. PMID- 8250413 TI - Researcher says RN care is key component to hospital competitiveness. Interview by Kathryn Scott. PMID- 8250414 TI - A career choice for collective bargaining. PMID- 8250415 TI - Nursing programs for career changers are on the rise. PMID- 8250416 TI - Study explores nurses' attitudes about career issues. PMID- 8250417 TI - A celebration of patriotism and courage. Vietnam Women's Memorial to be dedicated Nov. 11. PMID- 8250418 TI - Confidentiality crucial to nurse-patient relationship. PMID- 8250419 TI - Nurses discuss profession's role in assisted suicide and euthanasia. PMID- 8250420 TI - A night in the life of a nurse-midwife. PMID- 8250421 TI - Refresher course eases re-entry into nursing. PMID- 8250422 TI - Biofeedback can improve patient outcome. PMID- 8250423 TI - AAN forms grassroots network to mobilize nurses nationally. PMID- 8250424 TI - New York nurses rescue patients during fire. PMID- 8250425 TI - Southwestern communities respect rural RNs. PMID- 8250426 TI - Strengthening the states to be the heroes in reform. PMID- 8250427 TI - ANA, its work and the future of nursing. PMID- 8250428 TI - [Inflammatory bowel disease]. PMID- 8250429 TI - [Erythema nodosum in pediatric patients. A study of 22 cases]. AB - In this study we report the cases of 22 pediatric patients with nodal erythema. The predominance of this condition in male patients was clear. Etiological factors were determined in 77% of the patients. In our series, the principal cause was tuberculosis (36%). We would like to point out the etiological diversity in this small series of children: streptococcal infection, gastrointestinal infection with Salmonella enteritidis or Campylobacter jejuni, cat scratch disease, infectious mononucleosis, chronic hepatitis B, Crohn's disease and pharmacological (amoxycillin). In 22% of the cases no cause was found. PMID- 8250430 TI - [Umbilical colonization in normal newborns. A comparative study of 4 methods for umbilical antisepsis]. AB - We report the results of a prospective study in which the effectiveness of 70% alcohol (A), mercurochrome (M), a solution of alcohol and mercurochrome (AM) and 1% chlorhexidine (C) as umbilical antiseptics was tested on 311 normal newborn infants. Similar results were obtained by using A, M, or AM; however, when C was used umbilical colonization was significantly reduced as compared to when the other three antiseptic agents were used. The umbilical cords that had been scrubbed with A, M or AM were detached by the 8th day after birth, whereas those treated with C took 14 days to fall off. The use of chlorhexidine resulted in a marked increase in pure cultures positive for Gram negative bacteria (Proteus spp. and Pseudomonas spp.) which was not the case with the other three antiseptics tested. PMID- 8250431 TI - [Childhood kala-azar: the cases of a decade]. AB - The aim of our work was to review the epidemiological, clinical and laboratory features of the cases of Kala-azar in our hospital during a decade in order to outline data that might be useful in helping to make an earlier diagnosis and subsequently, earlier treatment. We report eighteen cases of visceral Leishmaniasis treated in our hospital between January 1981 and December 1990. The ages of the patients varied between five months and seven years. In our experience, medullar aspirate was the most effective diagnostic method. However, it this test is negative, specific serological tests should be included in the assessment of any pediatric inpatient with fever and splenomegaly. In our opinion, it is very important to keep this disease in mind, especially in non endemic areas. PMID- 8250432 TI - [The sudden infant death syndrome. II. Its prevention by home monitoring]. AB - Sudden infant death syndrome (SIDS) is a very dramatic situation with a very high incidence rate. Children with a high risk of SIDS can be identified by clinical/epidemiological scorings. The preventive usefulness of home monitoring is now a matter of study. Our experience is reported in this article. Thirty-nine infants at high risk were selected, although the monitoring program was finally achieved in only 36 cases. A cardiorespiratory impedance monitor (Centurion, Clinical Data, Boston, USA) was used. The tachycardia alarm was set at 200 b/min, bradycardia at 60 b/min and apnea > 20 seconds. The parents were trained in running the monitor and received technical support by phone during 24 hour per day. They filled out a daily record sheet with the observed anomalies. The mean time of monitoring was 8.2 months. After the withdrawal of the monitor, the parents answered a form with 26 questions. Twenty-four of the children needed, at least at some occasion, a light stimulation and 12 (33.3%) required a strong one. Meanwhile, the other 12 did not need any type of stimulus during monitoring. A simultaneous apnea/bradycardia alarm occurred in 17 cases. The home monitoring program was considered positive by the parents and only 4 families had technical difficulties. After the second week the fear and anxiety disappeared and only 1 family continued to be nervous throughout the monitoring program. The mean number of false alarms was 13.3/child, as opposed to 15.6 alarms due to apnea, 11.6 due to tachycardia and 0.9 due to apnea plus bradycardia. In conclusion, we think that home monitoring is easy, it reassures the families and their acceptance of the program is good. PMID- 8250433 TI - [The determination of IgG subclasses in healthy children by ELISA with monoclonal antibodies]. AB - The age at which the different IgG subclasses reach stable values has not been uniformly established to date. In this study the concentration of IgG1, IgG2, IgG3 and IgG4 has been determined in the serum of 164 children (from 1 to 14 years of age) and 20 healthy adults, by using a monoclonal antibody enzyme immunoassay. The geometric mean, standard deviation and coefficient of correlation were obtained. Results showed a gradual increase in the levels of the different subclasses throughout age. The time at which the adult concentrations were obtained varied and these adult levels were not necessarily the highest values. During the first year of life, the level of IgG1 is already high and by 6 7 years its serum concentration represented 90% of the adult IgG concentration. IgG2, IgG3 and IgG4 were initially detected at low levels. IgG2 increased rapidly until it reached a stable value at 9-10 years of age. Adult values were obtained at 10-11 years of age for IgG3 and 7-8 years of age for IgG4. PMID- 8250434 TI - [Acute meningococcal disease. Its prognostic assessment]. AB - We have studied 50 children affected with acute meningococcal disease (AMD). The ages of the children varied between 4 months and 12.58 years, with a mean age of 4.58 years. By using the shock state and DIC syndrome, both of which are indications of the severity of the illness, an evaluation of the discriminatory capacity was done with regard to significantly associate variables and 3 scores, Bjorvatn, Leclerc and PRISM, throughout 8 intervals within the first 48 hours of hospital treatment. We observed a very high survival rate (98%) associated with the early treatment for shock. Leukopenia and disseminated purpura were the best variables in order to discriminate shock and DIC, respectively. The greatest capacity for the diagnosis of the shock state and DIC syndrome were registered during the 0-6 hour period and the 0-12 hour period, respectively. The prognosis improved if the child remained alive 12 hours after the treatment had begun. PMID- 8250435 TI - [The prevalence of anemia in the children and adolescents of Madrid]. AB - Iron deficiency and iron deficiency anemia are very important disorders in childhood because, during this stage of life, the confluence of different factors can lead to iron deficiency. During childhood, there are rapid growth periods in which the need for iron increases and these needs are not always compensated for by a sufficient iron intake. We have studied a sample of 2,224 schoolchildren, both males and females, between the ages of 2 and 18 years, from five different schools in the Community of Madrid. The prevalence of anemia was studied by determining the hemoglobin, MCV and MCHC, whereas iron deficiency was assessed by measurement of serum iron. In our study population, the prevalence of iron deficiency was 4.94% and the rate of iron deficiency anemia was 0.94%. This disorder is predominant in 13 to 15 year olds (3.7% of the males and 3.53% of the females showed hemoglobin values below the reference range). In addition 6.73% of this group were iron deficient. The MCV was decreased in 2.69% of the subjects, after the exclusion of six children with thalassemia trait. PMID- 8250436 TI - [Elastase-alpha 1-antiprotease in neonatal sepsis]. AB - Elastase-alpha 1-proteinase inhibitor complex (E-alpha 1-PI) was evaluated in 682 blood samples from 516 newborn infants. They were divided into three groups: control (group 1; n = 99), non-infectious (group 2; n = 338) and infectious (group 3; n = 82). The plasma values of E-alpha 1-PI complex (median; minimum maximum, in microgram/L) were 180; 46-296 in group 1, 337; 40-2524 in group 2 and 954.5; 183-8160 in group 3. Significant differences were found between the groups (p < 0.001). E-alpha 1-PI complex in blood showed a sensitivity of 75%, specificity of 81.9%, positive predictive value of 30.6% and negative predictive value of 96.8% for the diagnosis of neonatal infection. PMID- 8250437 TI - [Berger's disease in children: its form of presentation, pathological anatomy and evolution in 22 cases]. AB - We have reviewed 22 cases of Berger's disease in children (glomerular nephritis with mesangial IgA deposits), all of which were diagnosed by renal biopsy between 1976 and the present time. We describe the clinical and pathological findings in these patients. In addition, we put special emphasis on the evolution of the disease in relationship to some of the parameters that have been reported in the literature as being related to a bad prognosis of glomerular function such as, massive proteinuria at the onset of the disease, histological classification, presence of deposits of IgM or fibrinogen derivatives and glomerular sclerosis. All of the patients started with hematuria, 21 of which had gross hematuria (95%). Fourteen patients (63%) showed proteinuria (2 of which also had a temporary nephrotic syndrome). Five children showed some transient decrease in glomerular filtration rate and another patient rapidly developed renal failure and then end stage renal disease. We were able to follow 15 children for 3 years: 8 (53%) still showed outbreaks of gross hematuria, 5 (33%) only had microhematuria and 2 (14%) showed no signs of hematuria. Four children (27%) still had proteinuria. The glomerular filtration rate was still normal in all but two children (one with rapid evolution to end stage renal disease and another with a glomerular filtration decrease of 20%). Ten children were followed for 6 to 13 years. After 6 years, 2 (20%) still showed outbreaks of gross hematuria, 1 (10%) still had proteinuria.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8250438 TI - [The medical care and vaccinal coverage of the children of Filipino mothers]. AB - We present data on the access to health care services and vaccination coverage of a sample of children born to Filipino mothers in the city of Barcelona (Spain). The sample was obtained by snowball sampling from the community reference centers for the identified migrant Filipino population. Access to public health care services is much lower for this population than for the whole population, which causes use of private services to compensate for this shortcoming. The causes are related to the residence and working status of a segment of this migrant population. Vaccination coverage for two and three year olds was 68% for measles, mumps and rubella, 52% with four doses for polio, diphtheria and tetanus and 48% with four doses for pertussis. Vaccination coverage is slightly higher for four and five year olds, with a coverage of 73.9% for measles, 69.5% for mumps and rubella and 73.9% with four doses for polio, diphtheria and tetanus, and 69.5% with four doses for pertussis. Vaccination coverage of 6 to 14 year old children is much lower. It is possible that real vaccination coverage is higher, but vaccination cards or records have been lost by the families. These results are much lower than global vaccination levels for the city, but comparable to those described in the inner city district where most of these children live. The causes of this situation, their implications and how to improve them are discussed. PMID- 8250439 TI - [The treatment of the Guillain-Barre syndrome in childhood by plasmapheresis]. AB - We have performed 18 plasmapheresis [correction of plasma exchange] sessions in a group of 5 patients diagnosed as having Guillain-Barre Syndrome. These patients were diagnosed according to clinical, analytical and electromyographic criteria. In three patients we observed an obvious clinical improvement during the treatment. In one of the patients there was no appreciable evolution. This patient suffered from acute lymphocytic leukemia. We were unable to evaluate the remaining patient since this child underwent posthypoxic encephalopathy secondary to cardiac arrest. On the basis of the results obtained, we advocate the treatment of this polyneuropathy by plasmapheresis [correction of plasma exchange]. PMID- 8250440 TI - [The long-term efficacy of a recombinant hepatitis B vaccine in newborns]. AB - A prospective evaluation of the immunogenicity, protective efficacy and safety of the recombinant hepatitis B vaccine associated to the immunoglobulin (IG) in newborns with high risk of infection by the hepatitis B virus was carried out. Two groups of newborns were used. The first group was formed by children with HBsAG carrier mothers (56 children) and the second group consisted of newborns of mothers negative for HBsAG, but having a high risk of infection (21 children). Within the first few hours of life, all of the newborns received a dose (20 mcg) or recombinant vaccine and 0.5 ml of IG. New doses of vaccine were administered at one and six months of life. At eight months of age, the anti-HBs seroconversion rate in children belonging to groups 1 and 2 was 100%, with an elevated anti-HBs titer and with no detected cases of positive HBsAG. On a long term basis (39 +/- 5 months), only 5.6% of the children present an anti-HBs titer < 10 mUI/ml. The recombinant hepatitis B vaccine associated to IG is immunogenic and provides efficient protection in newborns. Important side effects were not observed. PMID- 8250441 TI - [The Arnold-Chiari type-II malformation and apneas]. PMID- 8250442 TI - [A tick bite and lymphadenosis benigna cutis in childhood. Their treatment with josamycin]. PMID- 8250443 TI - [Paratesticular fibrous hamartoma in childhood. A report of a new case]. PMID- 8250444 TI - [Central core myopathy. A review of its complications]. PMID- 8250445 TI - [Congenital cystathioninuria. B6-dependent]. PMID- 8250446 TI - [Hepatoblastoma. A report of a case diagnosed in utero]. PMID- 8250447 TI - [Spontaneous perforation of the extrahepatic bile ducts in infancy]. PMID- 8250448 TI - [Iron deficiency anemia. Whose approach is better?]. PMID- 8250449 TI - [Breast feeding and the infant diet industry. 2 incompatible terms]. PMID- 8250450 TI - [The trend of the smoking habit among Bierzo adolescents]. PMID- 8250451 TI - Association of chronic nasal carriage of Staphylococcus aureus and higher relapse rates in Wegener granulomatosis. AB - OBJECTIVE: To examine possible risk factors for relapse, including chronic nasal carriage of Staphylococcus aureus and serial antineutrophil cytoplasmic antibody (ANCA) determinations in patients with Wegener granulomatosis. DESIGN: Observational cohort study. SETTING: Outpatient clinic at a university-affiliated hospital. PATIENTS: Consecutive patients (n = 71) with biopsy-proven Wegener granulomatosis who were seen during follow-up at the outpatient clinic from January 1988 to July 1991. Fourteen patients were ineligible or dropped out; 57 patients were analyzed. MEASUREMENTS: Serial ANCA determinations and swab cultures of both anterior nares for S. aureus taken at each visit every 4 to 6 weeks. Occurrence of infections and relapses of Wegener granulomatosis were identified according to strict, predefined criteria. RESULTS: Thirty-six of the 57 patients (63%; 95% CI, 49% to 76%) were found to be chronic nasal carriers of S. aureus (> or = 75% of nasal cultures positive for S. aureus). Proportional hazards regression analysis identified chronic nasal carriage of S. aureus (adjusted relative risk, 7.16; CI, 1.63 to 31.50), creatinine clearance above 60 mL.min-1 (adjusted relative risk, 2.94; CI, 1.27 to 6.67), and a history of previous relapses of Wegener granulomatosis (adjusted relative risk, 1.33; CI, 0.98 to 1.78) as independent risk factors for relapse. Twenty-two of 33 patients persistently or intermittently positive for ANCA had a relapse as opposed to only 1 of 21 persistently negative patients. Relapses of Wegener granulomatosis were not related to diagnosed infections. CONCLUSION: Chronic nasal carriage of S. aureus identifies a subgroup of patients with Wegener granulomatosis who are more prone to relapses of the disease, suggesting a role for S. aureus in its pathophysiology and a possible clue for treatment. PMID- 8250452 TI - Obesity as an adverse prognostic factor for patients receiving adjuvant chemotherapy for breast cancer. AB - OBJECTIVE: To determine whether obesity is an independent prognostic factor among women receiving adjuvant chemotherapy for lymph node-positive breast cancer and to determine how obesity relates to other commonly used prognostic indicators. DESIGN: Retrospective review of the clinical characteristics and clinical course of 735 patients with stages II and III primary breast cancer who were treated using three consecutive postoperative adjuvant chemotherapy protocols. Univariate and multivariate analyses were used to determine the prognostic implications of obesity defined by weight and height tables and body mass index. In addition, we analyzed the relation between obesity and other known prognostic indicators for patients with primary breast cancer. SETTING: A comprehensive cancer center. PATIENTS: 735 patients with lymph node-positive primary breast cancer who were treated using three consecutive fluorouracil-doxorubicin-cyclophosphamide containing adjuvant chemotherapy protocols and for whom complete data on weight, height, standard prognostic factors, and outcome were available. MAIN OUTCOME MEASUREMENTS: Disease-free and overall survival for the entire group and obese and nonobese subgroups. RESULTS: 24 percent of patients were more than 20% overweight. With a median follow-up of 10.7 years, the estimated 10-year, disease free rate for patients not more than 20% overweight was 54% (95% CI, 50% to 58%) compared with 40% (CI, 33% to 47%) for remaining patients classified as obese. Although obese patients tended to have somewhat less favorable prognoses based on standard prognostic criteria, a proportional-hazards regression model adjusting for other factors indicated that risk for disease recurrence among obese patients was 1.33 times that of the nonobese population (CI, 1.05 to 1.68). CONCLUSIONS: Obesity is an indicator of poor prognosis for patients with primary breast cancer even after the administration of adjuvant chemotherapy. The effect of dietary interventions to reduce body weight on the outcome of breast cancer therapy must be investigated. PMID- 8250453 TI - Prediction of long-term mortality in patients with rheumatoid arthritis according to simple questionnaire and joint count measures. AB - OBJECTIVE: To describe mortality over 15 years in a cohort of patients with rheumatoid arthritis, according to a simple questionnaire and joint count. DESIGN: A cohort study with 15 years of follow-up. SETTING: University hospital outpatient clinic. PATIENTS: A cohort of 75 patients with rheumatoid arthritis. MEASUREMENTS: Quantitative baseline measures: demographic, articular (joint counts), clinical, questionnaire, and physical measures, including modified questionnaire and joint count measures with substantially fewer items. RESULTS: Although few deaths were seen in the first 3 years after baseline, the standard mortality ratio over 15 years was 1.62, similar to findings in other series. Significant predictors of mortality included age, formal education level, joint count, activities-of-daily-living questionnaire scores, disease adjustment scores, morning stiffness, comorbid cardiovascular disease, grip strength, modified walking time, and button test. Five-year survival in patients with the poorest status according to these quantitative measures was 40% to 60%, comparable to expected survival at that time of patients with three-vessel coronary artery disease or with stage 4 Hodgkin disease. Simplified measures, including a count using only 28 joints and a questionnaire using only 8 activities of daily living, were similar to the more elaborate traditional measures for predicting mortality. CONCLUSION: Higher mortality rates in patients with rheumatoid arthritis are predicted by more severe clinical disease, as in other chronic diseases. Severe rheumatoid arthritis may be identified using quantitative functional status questionnaires and joint counts, which can be ascertained in about 10 to 15 minutes in any clinical setting. PMID- 8250454 TI - Incidence of cancer among men with the Felty syndrome. AB - OBJECTIVE: To estimate the incidence of cancer (especially lymphoproliferative malignancies) in patients with the Felty syndrome. DESIGN: A retrospective cohort study. SETTING: A computerized database of all discharge records for 1969 to 1990 from a Veterans Affairs hospital. PATIENTS: 906 men with a discharge diagnosis of the Felty syndrome. MEASUREMENTS: Standardized incidence ratios (SIR) (ratios of observed-to-expected events) estimated the risk for specific cancers. Hospital records confirmed the diagnoses of the Felty syndrome and cancer. RESULTS: We observed a twofold increase in total cancer incidence (137 patients; SIR = 2.09; 95% CI, 1.8 to 2.5). The risk for non-Hodgkin lymphoma (19 patients; SIR = 12.8, CI, 7.7 to 20.0) was much greater than the twofold increase in risk for lymphoma generally reported for rheumatoid arthritis. The risk for leukemia was increased but only within 5 years of the first hospitalization for the Felty syndrome, (13 patients; SIR = 7.67; CI, 4.1 to 13.1). CONCLUSION: The increased risk for non Hodgkin lymphoma after the Felty syndrome in our study is similar to the risk associated with the Sjogren syndrome and may reflect similar immunostimulatory mechanisms. PMID- 8250455 TI - Thrombolytic treatment and balloon angioplasty in chronic occlusion of the aortic bifurcation. AB - OBJECTIVE: To evaluate nonsurgical alternatives in reopening chronically occluded aortic bifurcation. DESIGN: Uncontrolled randomized study. SETTING: University affiliated referral center for vascular diseases. PATIENTS: Twenty-five of 39 consecutive patients with chronic aortoiliac disease including a totally occluded aortic bifurcation were found to be acceptable candidates for an aortobifemoral prosthetic graft. INTERVENTION: Patients were randomly assigned to receive either streptokinase or urokinase or recombinant tissue-type plasminogen activator (rt PA). In cases of successful thrombolysis and residual obstructions, subsequent balloon angioplasty was attempted. Prosthetic bypass grafting was done if thrombolytic treatment and balloon angioplasty failed. RESULTS: Complete lysis was achieved in 5 of 25 patients (20%). In 10 (40%) patients, lysis showed residual obstructions, which were reopened mechanically in 8 patients; 2 patients had extra-anatomical bypass grafts. Ten patients (40%) without thrombolysis had surgical aortobifemoral bypass grafts. Overall, recanalization and clinical improvement were achieved in 13 of 25 patients (52%) by thrombolytic therapy and subsequent balloon angioplasty. The recanalization rate did not differ among the different thrombolytic drugs. However, rt-PA therapy resulted in reopening after 4 days of treatment; streptokinase, after 6 days; and urokinase, after 9 days (P < 0.005). No major complications or deaths occurred. CONCLUSION: Thrombolytic treatment followed by balloon angioplasty may help avoid the need for aorto bifemoral prosthetic bypass grafting in more than 50% of patients with chronic aortoiliac disease. PMID- 8250456 TI - Insulin-like growth factor-1 therapy in diabetes: physiologic basis, clinical benefits, and risks. AB - PURPOSE: To review the effects of insulin-like growth factor-1 (IGF-1) and to discuss the clinical benefits and risks of using it in patients with diabetes. DATA SOURCES: Recent publications identified through a MEDLINE search using relevant keywords. STUDY SELECTION: Selected studies on the metabolic effects and kinetic mechanisms of in vitro IGF-1 and existing literature on the effects of IGF-1 on glucose and lipid metabolism in vivo with special emphasis on data from humans. DATA SYNTHESIS: The substantial stimulatory effect of IGF-1 on glucose uptake suggests that, in selected clinical situations, the drug may be an alternative to standard treatment of diabetes. Metabolic control in patients with extreme insulin resistance is improved after using IGF-1. Moreover, patients with type II (non-insulin-dependent) diabetes who receive IGF-1 have improved glucose tolerance and decreased hyperinsulinemia and hypertriglyceridemia. The complications associated with long-term administration of IGF-1 are unknown but might include the progression of certain neoplasms and diabetic complications, such as nephropathy and retinopathy. CONCLUSIONS: Insulin-like growth factor-1 may be a useful adjunct for treatment of diabetes and may even be the drug of choice in some patients with extreme insulin resistance who have metabolic emergencies. However, further data are needed to evaluate the risks and benefits of IGF-1 use in diabetes and in other states associated with impaired insulin action. PMID- 8250457 TI - Chemical pleurodesis for malignant pleural effusions. AB - PURPOSE: To provide information about available agents for chemical pleurodesis. DATA SOURCES: A MEDLINE search (1966 to October 1992) was conducted using the terms malignant pleural effusion and pleurodesis. STUDY SELECTION: All articles containing references to patients with recurrent, symptomatic, malignant pleural effusions treated with chemical pleurodesis were selected and reviewed for pleurodesis regimen, number of patients treated, success rate (complete response), and adverse effects. The agents studied included doxycycline, minocycline, tetracycline, bleomycin, cisplatin, doxorubicin, etoposide, fluorouracil, interferon-beta, mitomycin-c, Corynebacterium parvum, methylprednisolone, and talc. DATA EXTRACTION: Independent extraction by three observers. RESULTS: Studies including a total of 1168 patients with malignant pleural effusions were reviewed for efficacy of the pleurodesis agent and studies including 1140 patients were reviewed for toxicity. Chemical pleurodesis produced a complete response in 752 (64%) of 1168 patients. The success rate of the pleurodesis agents varied from 0% with etoposide to 93% with talc. Corynebacterium parvum, the tetracyclines, and bleomycin had success rates of 76%, 67%, and 54%, respectively. The most commonly reported adverse effects were pain (265 of 1140, 23%) and fever (220 of 1140, 19%). CONCLUSIONS: Doxycycline and minocycline, with success rates of 72% and 86%, respectively, appear to be effective tetracycline-replacement agents in the few patients studied. Talc appears to be the most effective and least expensive agent; however, insufflation has the disadvantages of the expense of thoracoscopy and the usual need for general anesthesia. Bleomycin appears to be less effective than talc and the tetracyclines and is substantially more expensive. PMID- 8250458 TI - Scorecard cardiovascular medicine. Its impact and future directions. AB - Public release of operator-specific data for cardiovascular procedures has set a new precedent, introducing the "scorecard" era. Justification exists for public disclosure, but the mechanics of appropriate data release are complex from a clinical, statistical, and logistic standpoint. Scorecard medicine may appropriately promote regionalization of medical centers and consolidation of services, but unless the process is directed effectively, it may impair the development of new treatments because of a more restrictive clinical practice environment. We propose revamping our current system to facilitate rapid and accurate access to outcome data in the local practice environment so that improvement in practice occurs on a voluntary basis rather than in response to punitive restrictions. A rational plan needs to be developed for dealing with high-risk patients, perhaps through compensation in regression models used to calculate expected outcomes, and for the start-up of novice physicians. Special provisions are needed to promote clinical research. Before procedures are done, it would be ideal to provide a full disclosure informed consent, whereby the physician reports operator-specific data and the patient's decision-making process is facilitated. Overall, appropriate implementation of scorecards could ultimately lead to a substantial improvement in the quality of U.S. cardiovascular medicine. PMID- 8250459 TI - Tuberculosis and the health care worker: a historical perspective. AB - Many hospital outbreaks of tuberculosis have occurred in recent years in the United States, resulting in tuberculosis infection and disease among health care workers and patients. Several hospital workers have died of nosocomially acquired multidrug-resistant tuberculosis. Assuring the safety of the health care worker with respect to tuberculosis has become an urgent priority. A review of the medical literature of the past 100 years reveals that our current view of tuberculosis care as an occupational hazard emerged only in the 1950s, after a fierce and extensive debate. Many authorities had felt that care of the tuberculous patient conferred a health advantage to the care provider. This paper reviews this debate and considers steps taken decades ago, before our current environmental interventions were available to ensure the safety of the health care worker. PMID- 8250461 TI - The ghost of Dr. Said. PMID- 8250460 TI - Thyrotoxic bone disease in women: a potentially reversible disorder. AB - OBJECTIVE: To measure changes in spinal and femoral neck bone mineral densities in patients treated for Graves thyrotoxicosis. DESIGN: Cohort study. SETTING: Tertiary care center. PATIENTS: Fifteen women with active Graves thyrotoxicosis. Six patients were premenopausal and nine were postmenopausal. All patients had evidence of thyrotoxicosis as indicated by a raised total serum thyroxine, suppressed serum thyroid-stimulating hormone, and an elevated technetium-99m pertechnetate thyroid scan. A control group of 15 healthy volunteers matched for age, sex, and menopausal status were followed during the same period. MEASUREMENTS: Bone mineral density was measured by dual-energy x-ray absorptiometry at baseline and after 12 months of antithyroid therapy when euthyroidism had been achieved. RESULTS: After 12 months of therapy, total serum thyroxine, total serum triiodothyronine, serum alkaline phosphatase, and serum bone Gla-protein activities had returned to normal in all patients (P < 0.001 for all comparison between initial and final biochemical measurements). During this interval, the mean lumbar spine bone mineral increased from an initial value of 1.01 g/cm2 to 1.07 g/cm2, an increase of 6.6% per year (95% CI, 3.6% to 9.6%) (P < 0.001 compared with controls). Increases in femoral neck (1.2%/y; CI, -2.1% to 4.5%; P = 0.2 compared with controls) and femoral trochanter bone mineral (3.2%/y; CI, 2.4% to 8.7%; P = 0.2 compared with controls) were not statistically significant. Using forced-entry multiple regression analysis, the severity of the thyrotoxicosis was independently associated with the percentage increment in lumbar spine bone mineral density after 12 months of antithyroid therapy. CONCLUSION: Effective treatment of Graves thyrotoxicosis was associated with increases in lumbar spine and femoral neck bone mineral. Although the changes in bone mineral were modest, our data suggest that thyrotoxic bone loss may be a reversible disorder. PMID- 8250462 TI - Transmission of hepatitis viruses. PMID- 8250463 TI - Health care reform is on the way: do we want to compete on quality? PMID- 8250464 TI - Cholesterol reduction and stroke. PMID- 8250465 TI - Arthritis, collagenous colitis, and discoid lupus. PMID- 8250466 TI - Quinine-induced pancytopenia and coagulopathy. PMID- 8250467 TI - Cognitive therapy for hypertension. PMID- 8250468 TI - Training more generalists. PMID- 8250469 TI - Training more generalists. PMID- 8250470 TI - Training more generalists. PMID- 8250471 TI - Risk assessment for worker exposure to agricultural pesticides: review of a workshop. PMID- 8250472 TI - Evaluation of agricultural worker exposure to, and absorption of, pesticides. AB - The evaluation of the occupational exposure of agricultural workers to pesticides is an integral part of the risk assessment for product safety and regulatory purposes. At present, there is no internationally accepted, harmonized approach to this exposure evaluation. This paper reviews the currently available methods for the measurement of exposure to, and absorption of, pesticides by workers involved in their use and in associated agricultural activities. Biological monitoring is recommended as the most precise means of estimating the absorbed dose of a pesticide, particularly if supported by human metabolism and pharmacokinetic data. A 'whole-body' sampling method is recommended for the measurement of dermal exposure. For concurrent exposure and biological monitoring a refined 'whole-body' method is recommended which involves the use of clothing representing that which workers normally wear under the prevailing conditions. A personal air sampling method is recommended for the measurement of inhalation exposure, to collect the 'inspirable' fraction (and/or, where necessary, vapour component) of pesticide. These ambient exposure monitoring methods may be conducted simultaneously with biological monitoring. Guidelines are proposed for the conduct of field studies to evaluate exposure to, and absorption of, pesticides to satisfy the requirements of regulatory authority and other organizations. PMID- 8250473 TI - Biological monitoring for pesticide absorption. AB - This paper reviews the development of biological monitoring as a method of assessing absorption of pesticides during occupational exposure. An approach to human volunteer studies to estimate the relationship between exposure to pesticides and excretion of metabolites is described and practical aspects of applying biological monitoring to field studies are considered. Case studies are used to illustrate some of the pitfalls of extrapolating from animals to man and to demonstrate the importance of human volunteer studies in establishing a sound basis for biological monitoring for pesticides. PMID- 8250474 TI - Predictive exposure modelling for pesticide registration purposes. AB - The health risk to the agricultural worker using pesticides is important for registration procedures since during mixing, loading and application he or she handles undiluted as well as diluted pesticide formulations. Furthermore, exposure may occur during daily activities in the crop (e.g. harvesting). Possible exposure may reach levels that lead to excessive health risks depending on the toxicity of the pesticide. In such cases measures to ensure safe handling have to be prescribed. In the present paper the published data, based on exposure occurring in agricultural practice, are considered for extrapolation to obtain surrogate exposure levels for risk assessment in registration procedures. The criteria necessary for the input of exposure data in data-bases, and the scope for extrapolation and its drawbacks, will be discussed. The various exposure models that have been described will be discussed and compared, and illustrated with a few cases. PMID- 8250475 TI - Risk assessments in the pesticide regulatory process. AB - The risk assessment process offers regulators, registrants, public interest groups and users a means of weighing apparent health effects of pesticides and other chemicals, and measuring their safety for use. This report contains the method for developing relative measures of safety for acute toxicity and expressions of risk for chronic toxicity and assesses-human exposure under a variety of extreme scenarios. Since workplace exposures usually exceed environmental exposures by two or more orders of magnitude, occupational experience with pesticides is critical to the regulatory process. Default assumptions, the adoption of conventional numbers in the absence of empirical data, for factors such as dermal absorption, metabolism, clearance and worker protective measures that seem conservative may weaken the reliability of the risk characterization process and may reduce the likelihood of recognizing significant risk. An exposure-based regulatory policy should feature measurements of absorbed dose validated using biomonitoring and margins of safety and estimates of risk that reflect human experience. This paper lists basic characteristics of unit processes of risk assessment and presents some strategies and pitfalls of the process as practiced in California in recent years. PMID- 8250476 TI - The role of occupational hygiene in exposure assessment by biological indices. A position paper from the Occupational Hygiene Committee of the International Congress on Occupational Health. PMID- 8250477 TI - Management of acoustic neuromas. AB - A survey of the management of acoustic neuromas in the broadest sense is given. The epidemiology and pathogenesis is described. The clinical development may take any imaginable course, and even though a slowly progressing unilateral hearing impairment, of the sensorineural type, is the main pattern of presentation, it is stressed that any type of symptom could appear in an acoustic neuroma patient. In order to make an early diagnosis a high index of suspicion must be the basic principle, with all patients with unilateral symptoms, not only hearing deficits, being suspected of suffering from acoustic neuromas until proven otherwise. All patients with hearing better than 70-80 dB should be subjected to ABR, and if there is any doubt about the normality of the response, the patient should proceed to MRI. Patients with poor hearing should go directly to MRI. The advantages and disadvantages of the different surgical approaches are described and an electric treatment algorithm is outlined: 1) All tumors measuring 25 mm or more on MRI are operated via the translabyrinthine approach. 2) All patients with PTA poorer than 30 dB, and SDS poorer then 70% are operated via the translabyrinthine approach. 3) Tumors less than 10 mm extrameatally, and PTA better than 30 dB and SDS better then 70% are removed via the middle fossa route. 4) Tumors measuring 10-25 mm and PTA better than 30 dB and SDS better than 70% are removed via the suboccipital route. PMID- 8250478 TI - [Surgical indications in angioma of the face]. AB - Hemangiomas are common vascular tumors of children. They always regress. Surgery takes place for late sequelae, such as cutaneous scarring and distortion of the facial structures. Early surgery is beneficial for the nasal cryano hemangioma, or for some eyelid hemangiomas. Venous malformations need to be manage from childhood to adulthood as they slowly worsen. Percutaneous embolization with Ethibloc and surgery are performed in order to minimize, or to improve the skin, muscle and bone distortion. Arteriovenous malformations are the most dangerous vascular anomalies of the face. Superselective arterial embolization prepares the surgical excision of the nidus. Difficulties in covering the excised area are frequently solved by flaps or cutaneous expansion. PMID- 8250479 TI - [Mycoses of the maxillary sinus. Apropos of 40 cases]. AB - Between 1985 and 1992, 40 patients with mycosis of the maxillary sinus were treated in the department at Hopital Saint Michel and of otolaryngology, head neck surgery at Hopital Saint Michel and their charts retrospectively analysed. The clinical signs, radiological findings and etiopathological hypothesis are described. The authors report their experience of functional endoscopic sinus surgery for treatment of mycosis of maxillary sinus. A high rate of postoperative successes, a low rate of complications, the recovery of a normal intrasinusal aerial flow and the easy endoscopic follow-up lead them to adverate the use FESS in primary treatment for mycosis of the maxillary sinus. PMID- 8250480 TI - [Value of auditory evoked potentials and electrocochleography in the diagnosis of small acoustic neuromas. Our experience on 131 cases]. AB - The auditory brainstem responses, in a series of 131 patients with surgically confirmed acoustic neuroma, are reported. Six groups have been individualized according to the value of the I-V interpeak latency, the synchronisation of the waveform shape and the need to perform an electrocochleography. If we exclude the sixteen cases of profoundly deafness, or cophosis, where ABR and ECoG don't permit the recording of waves, the electrophysiological investigations allowed to provide the diagnosis of retrocochlear lesion in 115 patients, for a rate of 98.26%. This high degree of sensitivity favorably with data reported in literature, and supports the reliability of this method for detection of acoustic neuroma. According to these results, our diagnosis workup of a patient suspected of having an acoustic neuroma, presenting normal ABR findings, is presented. PMID- 8250481 TI - [Thyroid oncology in everyday practice. Apropos of 105 cases]. AB - The charts of 105 patients treated for histologically documented nodular thyroid disease from 1976 to 1991 were analysed to determine the various diagnostic and therapeutic problems posed by the management of thyroid nodules in everyday clinical practice. The incidence of solitary nodular and multinodular disease was almost identical, 45.7% and 41% respectively. Radionucleid imaging was performed in only 61.9% (65) of patients and showed a distinct predominance of cold nodules as compared to multiheteronodular goiters, 67.7% (44/65) and 21.5% (14/65), respectively. Histological confirmation of diagnosis, obtained after fixation of the surgical specimen in paraffin, revealed a vast majority of benign versus malignant disease, 89.5% and 10.5%, respectively. 13.6% of cold nodules proved to be cancers. Extensive surgical resection either subtotal or total thyroidectomy, was performed in 41.6% of patients. Improvements in clinical practice and screening of goiters in endemic areas must contribute to a better definition of the surgical management of thyroid nodules. PMID- 8250482 TI - [Epidermoid carcinoma of the thyroid. A clinical study and review of the literature. Apropos of a case]. AB - Squamous cell carcinoma of the thyroid is very rare and fatal disease, with most patients dead within one year or less. The tumor may be derivated from metaplastic squamous elements of the thyroid or from transformation of papillary carcinoma. These lesions are usually uncompletely resectable. They also are radioresistant and chemotherapy inconclusive. PMID- 8250483 TI - [Pulsatile tinnitus and arteriovenous fistula. Apropos of a case]. AB - Objective pulsatile tinnitus is an unusual disease. Diagnosis, suspected by the interrogation, can be confirmed by vascular radiography. Treatments are discussed and can use surgical solution or interventional radiology. The authors describe one case of arteriovenous occipito-sinusal fistula revealed by pulsatile tinnitus and treated with success by means of embolisation. PMID- 8250484 TI - [Laser and glottis excision]. AB - The CO2 laser is being used in several department for primary treatment of limited vocal cord carcinoma. The surgical resection is not the same in several department. One speaks of vaporisation, laser excision, cordectomy, deep cordectomy. The purpose of this paper is to definitive 3 types of laser surgery for glottic cancer = partial cordectomy, cordectomy with exposition, enlarged cordectomy. This classification allow to know with precision the importance of the surgical resection in each case. PMID- 8250485 TI - [Pleomorphic adenoma of the nasal septum. Apropos of a case]. AB - Pleomorphic adenoma is the common benign tumor of glandular tissue occurring in the head neck region. The authors report a case localised in the nasal septum, which is an uncommon site for this tumor. Symptoms include unilateral nasal obstruction and intermittent nasal bleeding. Local surgical resection with histologic clear margins is the treatment of choice but long term follow-up is recommended. PMID- 8250486 TI - [How do I study laryngeal mobility?]. PMID- 8250487 TI - [Aging of cholinesterase after inhibition by organophosphates]. AB - Progressive inhibition of cholinesterases by organophosphates results from phosphorylation of the active-site serine. Phosphorylated cholinesterases may undergo a dealkylation reaction of the organophosphorus moiety leading to "aged" enzyme, i.e. conversion of the inhibited enzyme into a non-reactivable form. Aging occurs rapidly when the inhibitor is soman, a powerful nerve agent. This reaction promotes formation of a salt bridge between the protonated histidine of the active site catalytic triad and a negatively charged oxygen bound to the phosphorus atom. This reaction is accompanied by enzyme conformational and stability changes. In the research of compounds which retard or prevent the dealkylation reaction of organophosphate-cholinesterase conjugates, some allosteric effectors are relatively efficient by decreasing the velocity of the "aging" process. Knowledge of the three-dimensional structure of non-inhibited, inhibited and aged cholinesterases allows to understand the intimate mechanism of irreversible enzyme inhibition. Modeling of enzyme structure in the presence of effectors is essential to find out new therapeutic means against organophosphate poisoning. PMID- 8250488 TI - [Validation of the virus inactivation capacity of a procedure of human plasma albumin purification by chromatography]. AB - Almost the whole of the human plasma albumin preparations intended for clinical or biological uses is at present fractionated by cold ethanol precipitation technics based on the Cohn method. However, ion-exchange chromatographic processes have been recently developed. The aim of this work was the evaluation of the viral inactivation efficacy of an automated industrial chromatographic process allowing fractionation of 350 to 400 l of plasma per cycle (one precipitation step, three ion-exchange chromatography steps using the Spherodex Spherosil gels--Sepracor-IBF, Villeneuve la Garenne, France--and one pasteurization step. Three relevant viruses were selected for this validation study: the hepatitis B virus (HBV), the poliomyelitis virus and the human immunodeficiency virus (HIV). In order to comply with EEC and FDA regulatory documents, significant amounts of the tested viruses were spiked into the different fractions obtained during the various purification steps and their removal or inactivation during the subsequent step were determined. The validation study was performed under conditions which mimic the manufacturing process using fractions obtained during a semi-industrial fractionation. Moreover, residual viral infectivity was checked on after elution and washing of the columns for each chromatographic step. Results have pointed out: a) an overall reduction of 4.4 log 10 for HBV. Infectivity is judged by a combination of several markers and the DNA polymerase activity is the most affected particularly during the three ending purification steps; b) an overall reduction in virus titer > 10 log 10 for the poliomyelitis virus; c) an overall reduction in virus titer > 10 log 10 for HIV (four of the five steps have an important potential to inactivate this virus increasing the safety of the process). Moreover, no residual viral infectivities were detected after washing of the columns. In conclusion, this study showed the viral safety of human albumin purified using the chromatographic Spherodex-Spherosil process. As had been observed for fractionation by means of ethanol, the pasteurization step is necessary to ensure inactivation of two of the three viruses tested (HBV and poliomyelitis virus). This validation study allowed the preparation of a manufacturing and controls document for albumin and a marketing authorization has been issued by the "Laboratoire National de la Sante" (LNS, France). PMID- 8250489 TI - [Use of 7-amino-4-methylcoumarin for the spectrophotometric determination of nitrites]. AB - A new reagent of diazotisation, 7-amino-4-methylcoumarin (coumarin 120) is proposed for the spectrophotometric determination of small amounts of nitrite. To obtain molecules with an high absorptivity, the copulation with different aromatic amines is studied. N, N-dimethylaniline leads to the best results, particularly the reproducibility. Under the optimised conditions, a good linearity (between 10 micrograms/l and 200 micrograms/l) and a good reproducibility (2-3%) according to the amount of detected nitrites are obtained. PMID- 8250490 TI - [Phenotype profile of blood lymphocytes in bronchiolitis obliterans after lung transplantation]. AB - Bronchiolitis obliterans (BO) remains the major complication in long-term survivors with lung transplants, occurring in up to 30% of them. As a non invasive follow-up of lung recipients, we studied the phenotype of peripheral blood lymphocyte subsets. Using a flow cytometric analysis, we could define a specific pattern during BO. The most important findings were 1) disappearance of the CD19+ B cell population, despite normal or increased immunoglobulin blood levels; 2) marked decrease of the CD4+/CD8+ ratio; 3) dramatic increase in phenotypic cytotoxic effector T cells CD8+S6F1+ (MHC Class I-restricted allocytotoxicity) and CD3+CD4-CD8- (MHC Class I-non restricted allocytotoxicity); 4) marked increase of the CD4+CD29+ (helper/inducer T cell) to CD4+CD45RA+ (suppressor/inducer T cell) ratio associated with the loss of phenotypic suppressor/inducer CD4+CD45RA+ T cells. Moreover, we have shown that the maintenance triple immunosuppressive regimen that consisted of cyclosporin, prednisolone and azathioprine, did not affect the relative distribution of lymphocyte subsets, except for the CD3+CD4-CD8- cytotoxic subset that was slightly decreased under therapy. Thus, using a selected combination of lymphocyte membrane antigens, sequential prospective testing should be useful in the non-invasive follow-up of lung-transplanted patients to predict and halt the progressive course towards BO. PMID- 8250491 TI - [Luminescence and detection in liquid chromatography. I: Change of environment of analytes]. AB - The emissions of light by biorganisms or these obtained by alchemists were known since long time ago but the are used in analytical chemistry only when STOKES discovered that the intensity of this light was proportional to the quantity of the matter. The very large sensibilities reached, associated with the great separation's ability of the liquid chromatography allows to develop new processes for quantification of very low concentrations of luminescent or no luminescent molecules. Many pharmaceutical, biological toxicological environmental or alimentary applications show that it is possible in liquid chromatography to obtain a detection limit about the pico or femtomole when simple chemical process are used: direct potentialization of luminescence by addition of modifiers of the chemical environment of the analytes: solvents, cyclodextrins, surfactants, metallic ions, indirect potentialization of the luminescence by transfer of energy from an excited molecule: sensitized fluorescence and phosphorescence, excitation of the molecule by a chemical reaction or chemiluminescence. These aspects are emphasized and illustrated by some examples in three articles. PMID- 8250492 TI - [Biological safety in the laboratory. Biological risk, standardization and practice]. AB - Working with pathogens or genetically engineered micro-organisms is a potential hazard for scientists, health care workers, employees of pharmaceutical industry, and also for the environment. Carelessness, poor technique in the handling of infectious materials, needle sting or infectious aerosol exposure are the cause of laboratory acquired infection. Biosafety, corollary of biocontamination, is based on the combination of good microbiological techniques, facility design of the laboratory and safety equipment. So, four biosafety levels are appropriate for the operations performed and the hazard posed by the infectious agents. PMID- 8250493 TI - [Jean Choay (1923-1993)]. PMID- 8250494 TI - Proteinuria: pathophysiology, significance and recommendations for measurement in clinical practice. PMID- 8250495 TI - The effect of microalbuminuria on glycaemic control, serum lipids and haemostasis parameters in non-insulin-dependent diabetes mellitus. AB - Microalbuminuria is an important risk factor for cardiovascular disease in non insulin-dependent diabetes mellitus (NIDDM) patients although the pathogenic mechanism between microalbuminuria and cardiovascular disease has not yet been established. Microalbuminuria in insulin-dependent diabetes mellitus (IDDM) patients has been related to abnormalities in haemostasis, poor glycaemic control, disadvantageous alterations in the lipid spectrum and elevated concentrations of lipoprotein(a), another independent risk factor for cardiovascular disease. In this study the interrelations between microalbuminuria and metabolic control, lipoprotein(a), other blood lipids and several haemostasis parameters were studied in 96 NIDDM patients (50 women, 46 men). Forty-three patients showed microalbuminuria. No significant differences were found in blood lipids (Lp(a), serum cholesterol, low-density lipoprotein and high-density lipoprotein cholesterol and triglycerides), glycaemic control (HbA1c) and several haemostasis parameters (factor VII, VIII, fibrin monomer, thrombin-antithrombin III, D-dimer, tissue plasminogen activator antigen and plasminogen activator inhibitor-1) between the micro- and normoalbuminuric subgroups. In the microalbuminuric subgroup increased concentrations for plasminogen and alpha 2 antiplasmin were measured. In general, the presence of microalbuminuria was not associated with significant alterations in glycaemic control, blood lipids or haemostasis parameters in this group of 96 NIDDM patients. Further investigation is required to explain the excess cardiovascular mortality in patients with an elevated urinary albumin excretion rate. PMID- 8250496 TI - Plasma prekallikrein levels in patients with hepatocellular carcinoma and liver cirrhosis: a pilot study. AB - The concentration of plasma prekallikrein (PK) in five patients with hepatocellular carcinoma (HCC) has been measured and related to levels in 18 patients with liver cirrhosis (LC) and 30 healthy subjects. It was found that the mean PK level was significantly increased in patients with HCC, while patients with LC demonstrated lower concentrations, as compared with healthy subjects. The results indicate that PK might be useful in screening cirrhotic patients for HCC. Longitudinal studies of PK in a larger group of patients at risk of developing HCC are therefore recommended. PMID- 8250497 TI - Improved formulae for the judgement of intrathecally produced IgA and IgM in the presence of blood CSF barrier damage. AB - In order to discriminate between transsudated and intrathecally produced IgA and IgM in patients with blood cerebrospinal fluid (CSF) barrier (BCB) damage, we developed extended indices (IgA-EI and IgM-EI) for these immunoglobulins according to the general formula for extended index of a component X in CSF: X-EI = (CSF-X/serum-X)/(CSF-albumin/serum-albumin)a where a is a parameter specific for X. For IgA parameter a was found to be 1.15 and for IgM 1.9. A preliminary evaluation of IgA-EI and IgM-EI indicated lower false positive rates as compared to CSF IgA and IgM concentrations as well as 'conventional' IgA and IgM indices in cases with BCB damage, and essentially the same rates as for the hyperbolic formulae of Reiber and Felgenhauer. The importance of reliable sampling and analytical technique for IgM in CSF is discussed. PMID- 8250498 TI - Isoelectric focusing in an immobilized pH gradient for the detection of intrathecal IgG in cerebrospinal fluid: sensitivity and specificity for the diagnosis of multiple sclerosis. AB - Cerebrospinal fluid and serum from 192 patients was analysed for the presence of intrathecally synthesized oligoclonal IgG bands using isoelectric focusing in an immobilized pH gradient pH 7-10. The sensitivity of this method for the diagnosis of multiple sclerosis (MS) was 95% (21 of 22), or 75% if patients with suspected MS were included. The specificity for the diagnosis of MS was 98%, or 96% if the suspected MS patients were included. The very high specificity may be because the intrathecally synthesized oligoclonal IgG associated with MS is more alkaline than IgG from serum and is better detected in an immobilized alkaline pH gradient. PMID- 8250499 TI - Role of plasma phosphate measurements in detecting rickets of prematurity and in monitoring treatment. AB - Twice weekly plasma and urine measurements were made in 24 very low birth weight infants. Intravenous feeding was given whilst infants required respiratory support. Subsequently they received breast milk or formula milk with a vitamin D supplement of 400 U/day. Fourteen babies required intravenous feeding for more than 10 days. Six developed radiological rickets or severe osteoporosis, and these infants had plasma phosphate levels < 1.2 mmol/L on breast milk or < 1.8 mmol/L on formula milk. Babies without radiological rickets had plasma phosphate levels > 1.2 mmol/L on breast milk and > 1.8 mmol/L in all but one on formula milk. Successful treatment of rickets was associated with a rise in plasma phosphate to the above levels. Untimed urine calcium and phosphate concentrations expressed as creatinine ratios were not helpful in detecting babies with rickets, but may be useful in monitoring therapy. PMID- 8250500 TI - A blood spot assay for apo A1 and B lipoproteins and the apo B/A1 ratio. AB - A study of conditions for the elution of apo A1 and B lipoproteins from dried blood spots has led to the development of an apo B/A1 ratio assay with results for dried blood spots which are comparable with serum assays. This assay has been developed to be suitable for large scale population screening. The concept of measuring ratios for co-eluting blood constituents improves the accuracy and precision of blood spot assays and opens up the possibility that patients could take their own blood sample and send it to the laboratory by post. PMID- 8250501 TI - Hypertriglyceridaemia in a case of subcutaneous fat necrosis in a newborn. PMID- 8250502 TI - The effect of variations in haematocrit, mean cell volume and red blood cell count on reagent strip tests for glucose. PMID- 8250503 TI - A rapid selective high-performance liquid chromatography assay for cyclosporine. PMID- 8250504 TI - Use of a chemiluminescent acridinium ester label for cellular uptake assay of low density lipoprotein. PMID- 8250505 TI - Urine catecholamine excretion in long-term oral theophylline use. PMID- 8250506 TI - Successful treatment of short stature and delayed puberty in congenital magnesium losing kidney. AB - Bartter's syndrome is a well described but uncommon disease characterized by hypokalaemia and hyperplasia of the juxtaglomerular apparatus of the kidney. It may present in infancy with failure to thrive and muscle weakness; it commonly causes short stature. Lesions at different sites within the renal tubule have been proposed as the cause of the syndrome. However, the biochemical abnormalities in many cases can be explained by defective reabsorption of chloride in the ascending loop of Henle, with loss of sodium and water and a secondary increase in renin and aldosterone concentrations. Less severe cases have been described which present in adolescence and have tetany as a prominent feature. Primary renal loss of magnesium associated with potassium wasting has been described in such cases and it has been suggested that these can be distinguished from classical Bartter's syndrome by hypocalciuria. This less well characterized disease has been named Welt, Gitelman-Welt or Gitelman syndrome and may include deficient tubular reabsorption of chloride, but the sites of magnesium and potassium loss in the kidney are uncertain. We describe a patient with this syndrome who presented with short stature, delayed puberty and tetany and responded well to magnesium replacement. PMID- 8250507 TI - Measurement of dopamine, HVA and HMMA in untimed urine samples. PMID- 8250508 TI - Method comparison. PMID- 8250509 TI - What are the criteria to introduce new methods for determination of urinary N acetyl-beta-D-glucosaminidase activity? PMID- 8250510 TI - Secondary hyperparathyroidism. PMID- 8250511 TI - Laboratory diagnosis of phaeochromocytoma. PMID- 8250512 TI - Mononuclear blood cell magnesium. PMID- 8250513 TI - Serum creatinine: effect of delay in separation. PMID- 8250514 TI - Screening for microalbuminuria in a mixed ethnic diabetic clinic. PMID- 8250515 TI - HemoCue blood glucose analyser and haematocrit. PMID- 8250516 TI - [The value of laparoscopy in the staging of digestive system cancers]. AB - Sixty two patients with a gastrointestinal carcinoma were evaluated pre operatively by ultrasonography, CT scan and laparoscopy to seek liver metastases and/or peritoneal carcinomatosis. Performance levels of laparoscopy, ultrasonography and CT scan were comparable regarding the diagnosis of liver metastases. Laparoscopy was markedly better than CT scan and ultrasonography in the diagnosis of peritoneal carcinomatosis. Thus the investigation of choice for the detection of liver metastases is a single imaging technique. In contrast, if excision surgery is envisaged, all gastrointestinal carcinomas should be assessed by ultrasonography and/or CT scan and laparoscopy to definitely eliminate peritoneal carcinomatosis or small superficial liver metastases, in such a way as to avoid unnecessary laparotomy. PMID- 8250517 TI - [Alcoholic ketoacidosis. Apropos of a case with coma]. AB - Alcoholic ketoacidosis is rare and must be considered in the differential diagnosis of any alcoholic patient with acidosis. The pathogenesis is discussed on the basis of one case. It is easily reversible by simple saline plus glucose infusion. PMID- 8250518 TI - [Cancer of the anal canal. Combination radio-chemotherapy: fashion or necessity?]. PMID- 8250519 TI - [The use of probiotics in humans]. PMID- 8250520 TI - [Maintenance treatment of Crohn's disease using orally administered mesalazine (Pentasa). A controlled multicenter study. The Study Groups on the Treatment of Inflammatory Digestive Disorders]. 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) 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 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 on-going 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. CONCLUSION: 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.